PMID- 17395799 TI - Professional societies. ACS drops Iranian members, citing embargo. PMID- 17395798 TI - Genetics. Selfish genes could help disease-free mosquitoes spread. PMID- 17395800 TI - Clinical research. Testing a novel strategy against Parkinson's disease. PMID- 17395801 TI - Research funding. Canadian institutes get windfall without the bother of competition. PMID- 17395802 TI - Genetics. Sequencers of a famous genome confront privacy issues. PMID- 17395803 TI - Metagenomics. Massive microbial sequence project proposed. PMID- 17395804 TI - Uranium enrichment. Spinning a nuclear comeback. PMID- 17395805 TI - Phil Baran profile. Chemical high-flyer's strategy: take away the safety net. PMID- 17395806 TI - Plant pathology. Deadly wheat fungus threatens world's breadbaskets. PMID- 17395807 TI - Lunar and Planetary Science conference. Bringing Martian streaks and gullies down to Earth. PMID- 17395808 TI - Lunar and Planetary Science conference. Warped shorelines on a rolling Mars. PMID- 17395809 TI - Lunar and Planetary Science conference.Cold, cold bodies, warm hearts. PMID- 17395810 TI - Lunar and Planetary Science conference. Snapshots from the meeting. PMID- 17395811 TI - Wildlife population increases in Serengeti National Park. PMID- 17395813 TI - Coal-fired power plants: imprudent investments? PMID- 17395812 TI - HIV-malaria interactions: don't forget the drugs. PMID- 17395814 TI - Teaching. Opportunities to learn in America's elementary classrooms. PMID- 17395815 TI - Behavior. The ultimate ecosystem engineers. PMID- 17395816 TI - Medicine. Anticipating trouble from gene transcription. PMID- 17395818 TI - Climate change. Ice sheet stability and sea-level rise. PMID- 17395817 TI - Geophysics. Taking Earth's temperature. PMID- 17395819 TI - Atmospheric science. CO2 is not the only gas. PMID- 17395820 TI - Materials for aesthetic, energy-efficient, and self-diagnostic buildings. AB - It has become desirable to reduce the nonrenewable content and energy footprint of the built environment and to develop "smart buildings" that allow for inexpensive monitoring and self-diagnostic capabilities. Latest-generation embedded sensors, self-healing composites, and nanoscale and responsive materials may augur a time when buildings can substantially adjust to changing environmental and functional demands. However, faced with the legal liability resulting from unknown lifetime performance, designers and engineers have had little incentive to incorporate new material technologies into building designs. As efficiency issues become more acute, the potential for improvement in performance from new materials, together with partnerships between the materials science community and those entrusted with the design and engineering of the built environment, may offer real breakthroughs for the future. PMID- 17395821 TI - Scleractinian coral species survive and recover from decalcification. AB - Anthropogenic-driven accumulation of carbon dioxide in the atmosphere and projected ocean acidification have raised concerns regarding the eventual impact on coral reefs. This study demonstrates that skeleton-producing corals grown in acidified experimental conditions are able to sustain basic life functions, including reproductive ability, in a sea anemone-like form and will resume skeleton building when reintroduced to normal modern marine conditions. These results support the existence of physiological refugia, allowing corals to alternate between nonfossilizing soft-body ecophenotypes and fossilizing skeletal forms in response to changes in ocean chemistry. This refugia, however, does not undermine the threats to reef ecosystems in a high carbon dioxide world. PMID- 17395822 TI - Seismostratigraphy and thermal structure of Earth's core-mantle boundary region. AB - We used three-dimensional inverse scattering of core-reflected shear waves for large-scale, high-resolution exploration of Earth's deep interior (D'') and detected multiple, piecewise continuous interfaces in the lowermost layer (D'') beneath Central and North America. With thermodynamic properties of phase transitions in mantle silicates, we interpret the images and estimate in situ temperatures. A widespread wave-speed increase at 150 to 300 kilometers above the coremantle boundary is consistent with a transition from perovskite to postperovskite. Internal D'' stratification may be due to multiple phase-boundary crossings, and a deep wave-speed reduction may mark the base of a postperovskite lens about 2300 kilometers wide and 250 kilometers thick. The core-mantle boundary temperature is estimated at 3950 +/- 200 kelvin. Beneath Central America, a site of deep subduction, the D'' is relatively cold (DeltaT = 700 +/- 100 kelvin). Accounting for a factor-of-two uncertainty in thermal conductivity, core heat flux is 80 to 160 milliwatts per square meter (mW m(-2)) into the coldest D'' region and 35 to 70 mW m(-2) away from it. Combined with estimates from the central Pacific, this suggests a global average of 50 to 100 mW m(-2) and a total heat loss of 7.5 to 15 terawatts. PMID- 17395823 TI - Computational design of peptides that target transmembrane helices. AB - A variety of methods exist for the design or selection of antibodies and other proteins that recognize the water-soluble regions of proteins; however, companion methods for targeting transmembrane (TM) regions are not available. Here, we describe a method for the computational design of peptides that target TM helices in a sequence-specific manner. To illustrate the method, peptides were designed that specifically recognize the TM helices of two closely related integrins (alphaIIbbeta3 and alphavbeta3) in micelles, bacterial membranes, and mammalian cells. These data show that sequence-specific recognition of helices in TM proteins can be achieved through optimization of the geometric complementarity of the target-host complex. PMID- 17395824 TI - Ballistic electron microscopy of individual molecules. AB - We analyzed the transport of ballistic electrons through organic molecules on uniformly flat surfaces of bismuth grown on silicon. For the fullerene C60 and for a planar organic molecule (3,4,9,10-perylene-tetracarboxylic acid dianhydride), the signals revealed characteristic submolecular patterns that indicated where ballistic transport was enhanced or attenuated. The transport was associated to specific electronic molecular states. At electron energies of a few electron volts, this "scanning near-field electron transmission microscopy" method could be applied to various adsorbates or thin layers. PMID- 17395825 TI - Role of solvent-host interactions that lead to very large swelling of hybrid frameworks. AB - An unusually large expansion upon solvent adsorption occurs without apparent bond breaking in the network of a series of isoreticular chromium(III) or iron(III) diarboxylates labeled MIL-88A to D [dicarbox = fumarate (88A); terephthalate (1,4 BDC) (88B); 2,6-naphthalenedicarboxylate (2,6-NDC) (88C); and 4-4' biphenyldicarboxylate (4-4'-BPDC) (88D)]. This reversible "breathing" motion was analyzed in terms of cell dimensions (extent of breathing), movements within the framework (mechanism of transformation), and the interactions between the guests and the skeleton. In situ techniques show that these flexible solids are highly selective absorbents and that this selectivity is strongly dependent on the nature of the organic linker. PMID- 17395826 TI - Plastic deformation recovery in freestanding nanocrystalline aluminum and gold thin films. AB - In nanocrystalline metals, lack of intragranular dislocation sources leads to plastic deformation mechanisms that substantially differ from those in coarse grained metals. However, irrespective of grain size, plastic deformation is considered irrecoverable. We show experimentally that plastically deformed nanocrystalline aluminum and gold films with grain sizes of 65 nanometers and 50 nanometers, respectively, recovered a substantial fraction (50 to 100%) of plastic strain after unloading. This recovery was time dependent and was expedited at higher temperatures. Furthermore, the stress-strain characteristics during the next loading remained almost unchanged when strain recovery was complete. These observations in two dissimilar face-centered cubic metals suggest that strain recovery might be characteristic of other metals with similar grain sizes and crystalline packing. PMID- 17395827 TI - Permissive and instructive anterior patterning rely on mRNA localization in the wasp embryo. AB - The long-germ mode of embryogenesis, in which segments arise simultaneously along the anteriorposterior axis, has evolved several times in different lineages of the holometabolous, or fully metamorphosing, insects. Drosophila's long-germ fate map is established largely by the activity of the dipteran-specific Bicoid (Bcd) morphogen gradient, which operates both instructively and permissively to accomplish anterior patterning. By contrast, all nondipteran long-germ insects must achieve anterior patterning independently of bcd. We show that bcd's permissive function is mimicked in the wasp by a maternal repression system in which anterior localization of the wasp ortholog of giant represses anterior expression of the trunk gap genes so that head and thorax can properly form. PMID- 17395828 TI - Emergent biogeography of microbial communities in a model ocean. AB - A marine ecosystem model seeded with many phytoplankton types, whose physiological traits were randomly assigned from ranges defined by field and laboratory data, generated an emergent community structure and biogeography consistent with observed global phytoplankton distributions. The modeled organisms included types analogous to the marine cyanobacterium Prochlorococcus. Their emergent global distributions and physiological properties simultaneously correspond to observations. This flexible representation of community structure can be used to explore relations between ecosystems, biogeochemical cycles, and climate change. PMID- 17395829 TI - Cascading effects of the loss of apex predatory sharks from a coastal ocean. AB - Impacts of chronic overfishing are evident in population depletions worldwide, yet indirect ecosystem effects induced by predator removal from oceanic food webs remain unpredictable. As abundances of all 11 great sharks that consume other elasmobranchs (rays, skates, and small sharks) fell over the past 35 years, 12 of 14 of these prey species increased in coastal northwest Atlantic ecosystems. Effects of this community restructuring have cascaded downward from the cownose ray, whose enhanced predation on its bay scallop prey was sufficient to terminate a century-long scallop fishery. Analogous top-down effects may be a predictable consequence of eliminating entire functional groups of predators. PMID- 17395830 TI - Protein composition of catalytically active human telomerase from immortal cells. AB - Telomerase is a ribonucleoprotein enzyme complex that adds 5'-TTAGGG-3' repeats onto the ends of human chromosomes, providing a telomere maintenance mechanism for approximately 90% of human cancers. We have purified human telomerase approximately 10(8)-fold, with the final elution dependent on the enzyme's ability to catalyze nucleotide addition onto a DNA oligonucleotide of telomeric sequence, thereby providing specificity for catalytically active telomerase. Mass spectrometric sequencing of the protein components and molecular size determination indicated an enzyme composition of two molecules each of telomerase reverse transcriptase, telomerase RNA, and dyskerin. PMID- 17395831 TI - Regulation of hepatic stellate cell differentiation by the neurotrophin receptor p75NTR. AB - Differentiation of hepatic stellate cells (HSCs) to extracellular matrix- and growth factor-producing cells supports liver regeneration through promotion of hepatocyte proliferation. We show that the neurotrophin receptor p75NTR, a tumor necrosis factor receptor superfamily member expressed in HSCs after fibrotic and cirrhotic liver injury in humans, is a regulator of liver repair. In mice, depletion of p75NTR exacerbated liver pathology and inhibited hepatocyte proliferation in vivo. p75NTR-/- HSCs failed to differentiate to myofibroblasts and did not support hepatocyte proliferation. Moreover, inhibition of p75NTR signaling to the small guanosine triphosphatase Rho resulted in impaired HSC differentiation. Our results identify signaling from p75NTR to Rho as a mechanism for the regulation of HSC differentiation to regeneration-promoting cells that support hepatocyte proliferation in the diseased liver. PMID- 17395832 TI - Top-down versus bottom-up control of attention in the prefrontal and posterior parietal cortices. AB - Attention can be focused volitionally by "top-down" signals derived from task demands and automatically by "bottom-up" signals from salient stimuli. The frontal and parietal cortices are involved, but their neural activity has not been directly compared. Therefore, we recorded from them simultaneously in monkeys. Prefrontal neurons reflected the target location first during top-down attention, whereas parietal neurons signaled it earlier during bottom-up attention. Synchrony between frontal and parietal areas was stronger in lower frequencies during top-down attention and in higher frequencies during bottom-up attention. This result indicates that top-down and bottom-up signals arise from the frontal and sensory cortex, respectively, and different modes of attention may emphasize synchrony at different frequencies. PMID- 17395833 TI - Lactate sensitive transcription factor network in L6 cells: activation of MCT1 and mitochondrial biogenesis. AB - We hypothesized that in addition to serving as a fuel source and gluconeogenic precursor, lactate anion (La-) is a signaling molecule. Therefore, we screened genome-wide responses of L6 cells to elevated (10 and 20 mM) sodium-La- added to buffered, high-glucose media. Lactate increased reactive oxygen species (ROS) production and up-regulated 673 genes, many known to be responsive to ROS and Ca2+. The induction of genes encoding for components of the mitochondrial lactate oxidation complex was confirmed by independent methods (PCR and EMSA). Specifically, lactate increased monocarboxylate transporter-1 (MCT1) mRNA and protein expression within 1 h and cytochrome c oxidase (COX) mRNA and protein expression in 6 h. Increases in COX coincided with increases in peroxisome proliferator activated-receptor gamma coactivator-1alpha (PGC1alpha) expression and the DNA binding activity of nuclear respiratory factor (NRF)-2. We conclude that the lactate signaling cascade involves ROS production and converges on transcription factors affecting mitochondrial biogenesis. PMID- 17395834 TI - Out-of-pocket healthcare spending by the poor and chronically ill in the Republic of Korea. AB - OBJECTIVES: We estimated out-of-pocket health care spending and out-of-pocket spending burden ratio employing household equivalent income in the Republic of Korea. We examined variations in out-of-pocket spending, estimated out-of-pocket spending burden ratio employing household equivalent income, and identified factors associated with out-of-pocket spending. METHODS: We used the 1998 Korean National Health and Nutrition Survey, a nationally representative survey of 39,060 individuals. Our analyses examined out-of-pocket spending, out-of-pocket spending burden ratio, and health care use by socioeconomic status, insurance type, health care facility type, and chronic condition after we controlled for sociodemographic variables. RESULTS: The lowest income quintile spent 12.5% of their total income out-of-pocket on medical expenditures, which was 6 times that of the highest income quintile (2%). Among those with 3 or more chronic conditions, low-income Koreans had the highest out-of-pocket spending burden ratio (20%), which was 5 times the spending burden among high-income Koreans (4%). In multivariate analyses, the number of chronic conditions, insurance type, health care use, and health care facility type were associated with out-of-pocket spending. CONCLUSIONS: Out-of-pocket spending in Korea is regressive, because lower-income groups pay disproportionately more of their income compared with higher-income groups. Low-income individuals with multiple chronic conditions are particularly vulnerable. PMID- 17395835 TI - Differences in frequency of violence and reported injury between relationships with reciprocal and nonreciprocal intimate partner violence. AB - OBJECTIVES: We sought to examine the prevalence of reciprocal (i.e., perpetrated by both partners) and nonreciprocal intimate partner violence and to determine whether reciprocity is related to violence frequency and injury. METHODS: We analyzed data on young US adults aged 18 to 28 years from the 2001 National Longitudinal Study of Adolescent Health, which contained information about partner violence and injury reported by 11,370 respondents on 18761 heterosexual relationships. RESULTS: Almost 24% of all relationships had some violence, and half (49.7%) of those were reciprocally violent. In nonreciprocally violent relationships, women were the perpetrators in more than 70% of the cases. Reciprocity was associated with more frequent violence among women (adjusted odds ratio [AOR]=2.3; 95% confidence interval [CI]=1.9, 2.8), but not men (AOR=1.26; 95% CI=0.9, 1.7). Regarding injury, men were more likely to inflict injury than were women (AOR=1.3; 95% CI=1.1, 1.5), and reciprocal intimate partner violence was associated with greater injury than was nonreciprocal intimate partner violence regardless of the gender of the perpetrator (AOR=4.4; 95% CI=3.6, 5.5). CONCLUSIONS: The context of the violence (reciprocal vs nonreciprocal) is a strong predictor of reported injury. Prevention approaches that address the escalation of partner violence may be needed to address reciprocal violence. PMID- 17395836 TI - Socioeconomic status and risk for arsenic-related skin lesions in Bangladesh. AB - OBJECTIVES: Arsenic contamination of groundwater is a severe public health crisis in Bangladesh, where the population is exposed to arsenic in drinking water through tube wells used for groundwater collection. In this study, we explored the association between socioeconomic status and arsenic toxicity. METHODS: We used baseline data from 11438 men and women who were recruited into the Health Effects of Arsenic Longitudinal Study (HEALS), a prospective cohort study on the health effects of arsenic exposure in Bangladesh. We conducted analyses with logistic regression and generalized estimating equations. RESULTS: We found a strong dose-response association with all measures of arsenic exposure and skin lesions. We also found that the effect of arsenic was modified by land ownership on a multiplicative scale, with an increased risk among non-land owners associated with well water arsenic (P=.04) and urinary total arsenic concentrations (P=.03). CONCLUSIONS: Our study provides insight into potentially modifiable host characteristics and identifies factors that may effectively target susceptible population subgroups for appropriate interventions. PMID- 17395837 TI - Socioeconomic position, co-occurrence of behavior-related risk factors, and coronary heart disease: the Finnish Public Sector study. AB - OBJECTIVES: We examined the associations between socioeconomic position, co occurrence of behavior-related risk factors, and the effect of these factors on the relative and absolute socioeconomic gradients in coronary heart disease. METHODS: We obtained the socioeconomic position of 9337 men and 39,255 women who were local government employees aged 17-65 years from employers' records (the Public Sector Study, Finland). A questionnaire survey in 2000-2002 was used to collect data about smoking, heavy alcohol consumption, physical inactivity, obesity, and prevalence of coronary heart disease (myocardial infarction or angina diagnosed by a doctor). RESULTS: The age-adjusted odds of coronary heart disease were 2.1-2.2 times higher for low-income groups than high-income groups for both men and women, and adjustment for risk factors attenuated these associations by 13%-29%. There was no further attenuation with additional adjustment for the number of co-occurring risk factors, although socioeconomic disadvantage was associated with the co-occurrence of multiple risk factors. The absolute difference in coronary heart disease risk between socioeconomic groups could not be attributed to the measured risk factors. CONCLUSIONS: Interventions to reduce adult behavior-related risk factors may not completely remove socioeconomic differences in relative or absolute coronary heart disease risk, although they would lessen these effects. PMID- 17395838 TI - Predictors of low-intent and high-intent suicide attempts in rural China. AB - OBJECTIVES: Acts of suicide are heterogeneous with respect to level of intent. Data on correlates of intent are overwhelmingly from Western samples. We sought to identify correlates of low-intent and high-intent suicide attempts in China. METHODS: We compared 277 adult case patients who presented to the emergency department in a rural hospital because of attempted suicide to 277 community control individuals, pair-matched for age, gender, and location of residence. Attempted suicides were stratified into 3 levels of suicide intent: low, intermediate, and high. Paired logistic regression analyses were used to analyze the data. RESULTS: High-intent acts of suicide were associated with major depression, chronic stress, and a relative or associate who had a history of suicidal behavior. These correlates were not associated with low-intent acts of suicide. CONCLUSIONS: High-intent acts of suicide in China are the culmination of longstanding difficulties or symptoms including depression, chronic stress, and previous exposure to suicidal behavior. Prevention of high-intent acts of suicide should include a focus on these domains. PMID- 17395839 TI - Family history of diabetes, awareness of risk factors, and health behaviors among African Americans. AB - OBJECTIVES: We examined the role of family history of diabetes in awareness of diabetes risk factors and engaging in health behaviors. METHODS: We conducted a cross-sectional analysis of 1122 African American adults without diabetes who were participants in Project DIRECT (Diabetes Interventions Reaching and Educating Communities Together). RESULTS: After adjustment for age, gender, income, education, body mass index, and perceived health status, African Americans with a family history of diabetes were more aware than those without such a history of several diabetes risk factors: having a family member with the disease (relative risk [RR]=1.09; 95% confidence interval [CI]=1.03, 1.15), being overweight (RR=1.12; 95% CI=1.05, 1.18), not exercising (RR=1.17; 95% CI=1.07, 1.27), and consuming energy-dense foods (RR=1.10; 95% CI=1.00, 1.17). Also, they were more likely to consume 5 or more servings of fruits and vegetables per day (RR=1.31; 95% CI=1.02, 1.66) and to have been screened for diabetes (RR=1.21; 95% CI=1.12, 1.29). CONCLUSIONS: African Americans with a family history of diabetes were more aware of diabetes risk factors and more likely to engage in certain health behaviors than were African Americans without a family history of the disease. PMID- 17395840 TI - Social inequalities in perinatal mortality in Belo Horizonte, Brazil: the role of hospital care. AB - OBJECTIVES: We examined the contribution of hospital type and quality of care to perinatal mortality rates in the city of Belo Horizonte, Brazil. METHODS: We used a cohort study of all births (40953) and perinatal deaths (826) in Belo Horizonte in1999. After adjusting for maternal education and birthweight, we compared mortality rates according to hospital category--defined by a hospital's relation to the national Universal Public Health System (SUS)--and quality of care. We used the Wigglesworth Classification to examine perinatal deaths. RESULTS: After we controlled for birthweight and maternal education, the highest perinatal death rates were observed in private and philanthropic SUS-contracted hospitals (relative to private, non-SUS-contracted hospitals). Hospital quality was also directly associated with perinatal death rates. Mortality rates were especially high for normal-birthweight babies born in private SUS-contracted hospitals. Intrapartum asphyxia was the leading cause of preventable death. CONCLUSIONS: In a class-segregated health care system, such as Brazil's, disparities in quality of care between SUS-contracted and non-SUS-contracted hospitals contribute to the unacceptably high rates of perinatal mortality. PMID- 17395841 TI - Globalization, binational communities, and imported food risks: results of an outbreak investigation of lead poisoning in Monterey County, California. AB - OBJECTIVES: Although the burden of lead poisoning has decreased across developed countries, it remains the most prevalent environmental poison worldwide. Our objective was to investigate the sources of an outbreak of lead poisoning in Monterey County, California. METHODS: An investigation in 3 county health department clinics in Monterey County, California, was conducted between 2001 and 2003 to identify risk factors for elevated blood lead levels (> or = 10 microg/dL) among children and pregnant women. RESULTS: The prevalence of elevated blood lead levels was significantly higher in 1 of the 3 clinics (6% among screened children and 13% among prenatal patients). Risk factors included eating imported foods (relative risk [RR]=3.4; 95% confidence interval [CI]=1.2, 9.5) and having originated from the Zimatlan area of Oaxaca, Mexico, compared with other areas of Oaxaca (RR=4.0; 95% CI=1.7, 9.5). Home-prepared dried grasshoppers (chapulines) sent from Oaxaca were found to contain significant amounts of lead. CONCLUSIONS: Consumption of foods imported from Oaxaca was identified as a risk factor for elevated blood lead levels in Monterey County, California. Lead contaminated imported chapulines were identified as 1 source of lead poisoning, although other sources may also contribute to the observed findings. Food transport between binational communities presents a unique risk for the importation of environmental hazards [corrected] PMID- 17395842 TI - Changes in socioeconomic inequalities in cancer mortality rates among French men between 1968 and 1996. AB - OBJECTIVES: We investigated changes in socioeconomic inequalities in cancer mortality rates among men in France between 1968 and 1996. METHODS: We used a representative sample of 1% of the French population and studied 4 periods (1968 1974, 1975-1981, 1982-1988, and 1990-1996). Causes of death were obtained by direct linkage with the French national death registry. The socioeconomic position of men aged 35 to 59 years was measured by using the occupational class reported at the time of the census at the beginning of each period. Analyses were conducted for all cancers and specifically for lung, upper aerodigestive tract, esophageal, colorectal, and other cancers. RESULTS: In all analyses, we observed socioeconomic inequalities during the 4 periods considered; the inequalities increased between the first and the last period. Most of the total increase occurred between 1968 and 1981, and inequalities remained stable thereafter. Inequalities were larger when men out of the labor force were included in the analysis. The strongest increase in socioeconomic inequalities over time was observed for upper aerodigestive tract cancer. CONCLUSIONS: Although cancer mortality rates have decreased, substantial socioeconomic inequalities in cancer mortality among men remain. PMID- 17395843 TI - Effects of the Office of National Drug Control Policy's Marijuana Initiative Campaign on high-sensation-seeking adolescents. AB - OBJECTIVES: We evaluated the effects of the Marijuana Initiative portion of the Office of National Drug Control Policy's National Youth Anti-Drug Media Campaign on high-sensation-seeking and low-sensation-seeking adolescents. METHODS: Personal interviews were conducted via laptop computers with independent monthly random samples of 100 youths from the same age cohort in each of 2 moderate-sized communities over 48 months (April 1999-March 2003) of the campaign, including the critical first 6 months of the 9-month initiative. The start of the initiative was treated as an "interruption" in time-series analyses of the combined community sample. RESULTS: The Marijuana Initiative reversed upward developmental trends in 30-day marijuana use among high-sensation-seeking adolescents (P<.001) and significantly reduced positive marijuana attitudes and beliefs in this at risk population. Use of control substances was not affected. As expected, low sensation-seeking adolescents had low marijuana-use levels, and the campaign had no detectable effects on them. Other analyses indicated that the initiative's dramatic depiction of negative consequences of marijuana use was principally responsible for its effects on high-sensation-seeking youths. CONCLUSIONS: Substance use prevention campaigns can be effective within an approach using dramatic negative-consequence messages targeted to high-sensation seekers. PMID- 17395844 TI - Trends in characteristics and country of origin among foreign-trained nurses in the United States, 1990 and 2000. AB - OBJECTIVES: We describe long-term trends in the characteristics of foreign trained new entrants to the registered nurse (RN) workforce in the United States. METHODS: Using the 1990 and 2000 US Census 5% Public Use Microdata Sample files, we compared trends in characteristics of US- and foreign-trained new entrants to the RN labor force (n=40827) and identified trends in the country of origin of the foreign-trained new entrants. RESULTS: Foreign-trained RNs grew as a percentage of new entrants to the RN workforce, from 8.8% in 1990 to 15.2% in 2000. Compared with US-trained RNs, foreign-trained RNs were 3 times as likely to work in nursing homes and were more likely to have earned a bachelor's degree. In 2000, 21% of foreign-trained RNs originated from low-income countries, a doubling of the rate since 1990. CONCLUSIONS: Foreign-trained RNs now account for a substantial and growing proportion of the US RN workforce. Our findings suggest foreign-trained RNs entering the United States are not of lower quality than US trained RNs. However, growth in the proportion of RNs from low-income countries may have negative consequences in those countries. PMID- 17395845 TI - A comparative health survey of the inhabitants of Roma settlements in Hungary. AB - OBJECTIVES: We compared the health of people living in Roma settlements with that of the general population in Hungary. METHODS: We performed comparative health interview surveys in 2003 to 2004 in representative samples of the Hungarian population and inhabitants of Roma settlements. RESULTS: In persons older than 44 years, 10% more of those living in Roma settlements reported their health as bad or very bad than did those in the lowest income quartile of the general population. Of those who used any health services, 35% of the Roma inhabitants and 4.4% of the general population experienced some discrimination. In Roma settlements, the proportion of persons who thought that they could do much for their own health was 13% to 15% lower, and heavy smoking and unhealthy diet were 1.5 to 3 times more prevalent, than in the lowest income quartile of the general population. CONCLUSIONS: People living in Roma settlements experience severe social exclusion, which profoundly affects their health. Besides tackling the socioeconomic roots of the poor health of Roma people, specific public health interventions, including health education and health promotion programs, are needed. PMID- 17395846 TI - The Latino paradox in neighborhood context: the case of asthma and other respiratory conditions. AB - OBJECTIVES: Evidence indicates that foreign-born Latinos have a health advantage compared with US-born persons of the same socioeconomic status. An explanation for this paradox has remained elusive. We examined the extent to which this paradox exists for the prevalence of asthma and other respiratory conditions. We then explored the role of neighborhood social context in understanding any observed advantage. We invoked theories of social organization, collective efficacy, and the urban ethnic enclave. METHODS: We combined data from the Project on Human Development in Chicago Neighborhoods Community Survey with 2 other data sources and used hierarchical generalized linear modeling techniques. RESULTS: We found a distinctly graded effect for asthma and other breathing problems among foreign-born Latinos, depending on community composition. Foreign born Latinos embedded in a neighborhood that had a high percentage of foreign born residents experienced a significantly lower prevalence of asthma and other breathing problems; those in communities that had a low percentage of foreign born residents had the highest prevalence overall (even when compared with African Americans). CONCLUSIONS: Foreign-born Latinos have a respiratory health advantage only in enclave-like settings. Contexts such as these may provide the cohesiveness critical for effective prevention. PMID- 17395847 TI - The relationship between neighborhood characteristics and self-rated health for adults with chronic conditions. AB - OBJECTIVES: We sought to determine whether the association between neighborhood characteristics and health differs for people with and without a chronic condition. METHODS: We analyzed data from 2536 adults from the Los Angeles Family and Neighborhood Survey and evaluated the relationship between the presence of a chronic condition at the individual level, neighborhood socioeconomic status (SES), and self-rated health. We constructed multilevel models to evaluate the relationship between the neighborhood SES index and self-rated health for people with and without chronic conditions, after adjustment for other individual characteristics. RESULTS: Having a chronic condition was associated with substantially poorer self-rated health among participants in a deprived area than among those in a more advantaged area. CONCLUSIONS: Residence in a disadvantaged neighborhood may be associated with barriers to the management of a chronic condition. Further work is needed to identify the specific characteristics of disadvantaged areas associated with poorer self-rated health for adults with chronic conditions. PMID- 17395848 TI - Generating political priority for maternal mortality reduction in 5 developing countries. AB - I conducted case studies on the level of political priority given to maternal mortality reduction in 5 countries: Guatemala, Honduras, India, Indonesia, and Nigeria. Among the factors that shaped political priority were international agency efforts to establish a global norm about the unacceptability of maternal death; those agencies' provision of financial and technical resources; the degree of cohesion among national safe motherhood policy communities; the presence of national political champions to promote the cause; the deployment of credible evidence to show policymakers a problem existed; the generation of clear policy alternatives to demonstrate the problem was surmountable; and the organization of attention-generating events to create national visibility for the issue. The experiences of these 5 countries offer guidance on how political priority can be generated for other health causes in developing countries. PMID- 17395849 TI - Race and the politics of polio: Warm Springs, Tuskegee, and the March of Dimes. AB - The Tuskegee Institute opened a polio center in 1941, funded by the March of Dimes. The center's founding was the result of a new visibility of Black polio survivors and the growing political embarrassment around the policy of the Georgia Warm Springs polio rehabilitation center, which Franklin Roosevelt had founded in the 1920s before he became president and which had maintained a Whites only policy of admission. This policy, reflecting the ubiquitous norm of race segregated health facilities of the era, was also sustained by a persuasive scientific argument about polio itself: that Blacks were not susceptible to the disease. After a decade of civil rights activism, this notion of polio as a White disease was challenged, and Black health professionals, emboldened by a new integrationist epidemiology, demanded that in polio, as in American medicine at large, health care should be provided regardless of race, color, or creed. PMID- 17395850 TI - Self-rated health trajectories in the United States and the United Kingdom: a comparative study. AB - OBJECTIVES: We reviewed literature on comparative social policy and life course research and compared associations between health and socioeconomic circumstances during an 11-year period in the United States and the United Kingdom. METHODS: We obtained data from the US Panel Study of Income Dynamics and the British Household Panel Survey (1990-2002). We used latent transition analysis to examine change in self-rated health from one discrete state to another; these health trajectories were then associated with socioeconomic measures at the beginning and at the end of the study period. RESULTS: We identified good and poor latent health states, which remained relatively stable over time. When change occurred, decline rather than improvement was more likely. UK populations were in better health compared with US populations and were more likely to improve over time. Labor market participation was more strongly associated with good health in the United Kingdom than in the United States. CONCLUSIONS: National policies and practices may be keeping more US workers than UK workers who are in poor health employed, but British policies may give UK workers the chance to return to better health and to the labor force. PMID- 17395851 TI - Associations between body composition, anthropometry, and mortality in women aged 65 years and older. AB - OBJECTIVES: We examined the relation between measures of body size and mortality in a predominantly White cohort of 8029 women aged 65 years and older who were participating in the Study of Osteoporotic Fractures. METHODS: Body composition measures (fat and lean mass and percentage body fat) were calculated by bioelectrical impedance analysis. Anthropometric measures were body mass index (BMI; kg/m2) and waist circumference. RESULTS: During 8 years of follow-up, there were 945 deaths. Mortality was lowest among women in the middle of the distribution of each body size measure. For BMI, the lowest mortality rates were in the range 24.6 to 29.8 kg/m2. The U-shaped relations were seen throughout the age ranges included in this study and were not attributable to smoking or measures of preexisting illness. Body composition measures were not better predictors of mortality than BMI or waist girth. CONCLUSIONS: Our results do not support applying the National Institutes of Health categorization of BMI from 25 to 29.9 kg/m2 as overweight in older women, because women with BMIs in this range had the lowest mortality. PMID- 17395852 TI - Husbands' involvement in housework and women's psychosocial health: findings from a population-based study in Lebanon. AB - OBJECTIVES: We examined the association between husbands' involvement in housework and the psychosocial health of their wives using data on married couples living in poor neighborhoods in Beirut, Lebanon. METHODS: Data were derived from a cross-sectional survey of 2797 households; 1652 married couples and their families were included in the analysis. An index of husbands' relative involvement in housework was constructed from 25 items focusing on division of housework activities. Logistic regression was used to assess associations between husbands' involvement in housework and wives' self-rated mental health status, marital dissatisfaction, and unhappiness. RESULTS: Husbands' involvement in housework was negatively associated with wives' psychological distress, marital dissatisfaction, and overall unhappiness after adjustment for relevant risk factors. In comparison with wives whose husbands were highly involved in housework, wives whose husbands were minimally involved were 1.60 times more likely to be distressed, 2.96 times more likely to be uncomfortable with their husbands, and 2.69 times more likely to be unhappy. CONCLUSIONS: Our results showed a significant association between husbands' involvement in housework and their wives' psychosocial health. PMID- 17395853 TI - Agent-based modeling: use with necessary caution. PMID- 17395854 TI - Topological determinants and consequences of adventitial responses to arterial wall injury. AB - Arteries are composed of 3 concentric tissue layers which exhibit different structures and properties. Because arterial injury is generally initiated at the interface with circulating blood, most studies performed to unravel the mechanisms involved in injury-induced arterial responses have focused on the innermost layer (intima) rather than on the outermost adventitial layer. In the present review, we focus on the involvement of the adventitia in response to various types of arterial injury leading to vascular remodeling. Physiologically, soluble vascular mediators are centrifugally conveyed by mass transport toward the adventitia. Moreover, in pathological conditions, neomediators and antigens can be generated within the arterial wall, whose outward conveyance triggers different patterns of local adventitial response. Adventitial angiogenesis, immunoinflammation, and fibrosis sequentially interact and their net balance defines the participation of the adventitial response in arterial pathology. In the present review we discuss 4 pathological entities in which the adventitial response to arterial wall injury participates in arterial wall remodeling. Hence, the adventitial adaptive immune response predominates in chronic rejection. Inflammatory phagocytic cell recruitment and initiation of a shift from innate to adaptive immunity characterize the adventitial response to products of proteolysis in abdominal aortic aneurysm. Adventitial sprouting of neovessels, leading to intraplaque hemorrhages, predominates in atherothrombosis. Adventitial fibrosis characterizes the response to mechanical stress and is responsible for the constrictive remodeling of arterial segments and initiating interstitial fibrosis in perivascular tissues. These adventitial events, therefore, have an impact not only on the vessel wall biology but also on the surrounding tissue. PMID- 17395855 TI - Cyclic strain regulates the Notch/CBF-1 signaling pathway in endothelial cells: role in angiogenic activity. AB - OBJECTIVE: The purpose of this study was to determine the effect of cyclic strain on Notch signaling in endothelial cells. METHODS AND RESULTS: Exposure of human endothelial cells (ECs) to cyclic strain (10%) resulted in temporal upregulation of Notch receptors (1 and 4) at the mRNA and protein level. Cyclic strain significantly increased EC network formation on Matrigel (an index of angiogenesis); network AU=775+/-127 versus 3928+/-400 for static and strained ECs, respectively. In addition, Angiopoietin 1 (Ang1), Tie1, and Tie2 expression were increased and knockdown of Ang1/Tie1,2 by siRNAs decreased cyclic strain induced network formation. Knockdown of Notch 1 and 4 by siRNA, or inhibition of Notch mediated CBF-1/RBP-Jk regulated gene expression by RPMS-1, caused a significant decrease in cyclic strain-induced network formation and in Tie1 and Tie2 mRNA expression. Notch 1 or Notch 4 siRNA, but not RPMS-1, inhibited cyclic strain-induced Ang1. Constitutive overexpression of Notch IC resulted in increased network formation, and Ang1 and Tie2 mRNA expression, under both static and strain conditions. CONCLUSIONS: These data suggest that cyclic strain stimulated EC angiogenesis is mediated in part through a Notch-dependent, Ang1/Tie2 signaling pathway. This pathway may represent a novel therapeutic target for disease states in which hemodynamic force-induced angiogenesis occurs. PMID- 17395856 TI - Liposome-encapsulated hemoglobin reduces the size of cerebral infarction in the rat: evaluation with photochemically induced thrombosis of the middle cerebral artery. AB - BACKGROUND AND PURPOSE: Liposome-encapsulated hemoglobin (LEH; TRM-645) is a novel oxygen (O(2)) carrier with a lower O(2) affinity (P(50)O(2)=40 mm Hg) than red blood cells. In contrast to cell-free hemoglobin, encapsulation prevents hemoglobin extravasation, whereas its subcellular size (230 nm) may improve O(2) delivery and limit the severity of cerebral infarction. METHODS: The extent of cerebral infarction was determined 24 hours after photochemically induced thrombosis of the middle cerebral artery from the integrated area of infarction detected by triphenyltetrazolium chloride staining in rats receiving no treatment, 10 mL/kg of LEH, homologous blood, empty liposomes, or saline. To develop a dose-response relationship, LEH dose was reduced from 10 mL/kg to 2 mL/kg, 0.4 mL/kg, and 0.08 mL/kg. RESULTS: Infarction extent was significantly suppressed in rats receiving LEH as compared with animals receiving no infusion, saline, empty liposome, or transfusion in the cortex but not in the basal ganglia, where all had similar degrees of damage. The dose-response relationship revealed that as little as 2 mL/kg of LEH was protective, whereas the total blood O(2) content, hemoglobin level, and transfusion and/or infusion of empty liposomes or saline were not effective. CONCLUSIONS: Our results suggest that LEH significantly reduces the area of infarction in the cortex but not in basal ganglia after photochemically induced thrombosis of the middle cerebral artery in the rat. PMID- 17395857 TI - Combining aspirin with oral anticoagulant therapy: is this a safe and effective practice in patients with atrial fibrillation? PMID- 17395858 TI - So, what's new? PMID- 17395859 TI - SRC kinase inhibition improves acute outcomes after experimental intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: The mechanisms by which intracerebral hemorrhages produce changes of blood flow and metabolism, cell death, and behavioral abnormalities are complex. In this study, we begin to test the hypothesis that intracerebral hemorrhage activates Src kinases that phosphorylate other molecules to produce cell injury and behavioral deficits after intracerebral hemorrhage (ICH). METHODS: ICH was produced in adult Sprague Dawley rats by direct injection of autologous blood (50 microL) into striatum. Src kinase activity, glucose hypermetabolic areas around the ICH, TUNEL-stained cells, and apomorphine-induced rotational behaviors were assessed in animals with ICH pretreated with the Src kinase inhibitor, PP1, or with vehicle. RESULTS: PP1 (3 mg/kg) blocked increases of Src kinase activity (5-fold) at 3 hours after ICH. PP1 also blocked the areas of glucose hypermetabolism and decreased the numbers of TUNEL-stained cells surrounding the ICH at 24 hours. Finally, apomorphine-induced (1 mg/kg) rotation at 24 hours after ICH was markedly attenuated by previous treatment with PP1 (3 mg/kg intraperitoneal). CONCLUSIONS: PP1 decreases Src kinase activation, glucose metabolic activation, cell death, and behavioral abnormalities after ICH in striatum of adult rats. It is hypothesized that intracerebral hemorrhage, possibly via thrombin activation of protease-activated receptors, activates Src that phosphorylates NMDA receptors, matrix metalloproteinases, and other proteins that mediate injury after ICH. PMID- 17395860 TI - Na+/Ca2+ exchanger maintains ionic homeostasis in the peri-infarct area. AB - BACKGROUND AND PURPOSE: A prominent feature of cerebral ischemia is the excessive intracellular accumulation of both Na(+) and Ca(2+) ions, which results in subsequent cell death. The plasma membrane Na(+)/Ca(2+) exchanger (NCX), regulates the distribution of these ions acting either in the forward mode or in its reverse mode and it can play a critical role in brain ischemia. However, it is unclear whether the activity of NCX leads to detrimental or beneficial effects. METHODS: Extracellular field potentials and whole-cell patch clamp recordings were obtained from rat corticostriatal brain-slice preparations in the peri-infarct area 24 hours after the permanent middle cerebral artery occlusion. Ischemia was induced in rats by permanent middle cerebral artery occlusion. RESULTS: Bepridil, an inhibitor of NCX, reduced in a concentration-dependent manner (IC(50)=68 micromol/L) the field potential amplitude recorded from the peri-infarct area of corticostriatal slices. Conversely, no change was observed in sham-operated animals. The effect of bepridil was mimicked by 5-(N-4 chlorobenzyl)-2',4'-dimethylbenzamil (CB-DMB) (IC(50)=6 micromol/L), a more selective inhibitor of NCX. In whole-cell patch clamp experiments, bepridil and CB-DMB caused an inward current in spiny neurons recorded from the peri-infarct area but not in the same cells recorded from controls. Interestingly, cholinergic interneurons recorded from the striatal peri-infarct area did not develop an inward current after the application of NCX inhibitors, suggesting that the electrophysiological alterations induced by NCX inhibition are cell-type specific. Bepridil and CB-DMB also induced a suppression of excitatory synaptic currents in most of spiny neurons recorded from the peri-infarct area. This effect was not coupled to a significant change of paired-pulse facilitation suggesting a postsynaptic site of action. CONCLUSIONS: Our data indicate that NCX plays a critical role in the maintenance of ionic homeostasis in the peri-infarct area. PMID- 17395861 TI - Reduced medication access: a marker for vulnerability in US stroke survivors. AB - BACKGROUND AND PURPOSE: Medication access is crucial to secondary stroke prevention. We assessed medication access and associated barriers to care across region and time in a national sample of US stroke survivors. METHODS: Among all 5840 black or white stroke survivors aged > or =45 years responding to the National Health Interview Survey years 1997 to 2004, we examined inability to afford medications within the last 12 months across region (Northeast, Midwest, West, South) and time. With logistic regression, we adjusted associations between medication inaffordability and region and time for age, sex, race, neurological disability, comorbidity, health status, insurance, income and out-of-pocket medical expenses. RESULTS: In 2004, approximately 76,000 US stroke survivors were unable to afford medications. Lower medication affordability was reported among stroke survivors who were <65 years old, black, female, had high comorbidity or low health status. Compared with stroke survivors able to afford medications, those unable more frequently reported lack of transportation (15% versus 3%; P<0.001), no health insurance (16% versus 3%; P<0.001), no usual place of care (6% versus 2%; P=0.001), income <$20,000 (66% versus 40%; P<0.001) and out-of pocket medical expenses > or =$2000 (35% versus 25%; P<0.001). From 1997 to 2004, inability to afford medications increased significantly from 8.1% to 12.7% (P(trend)=0.01) overall and increased in all US regions except the Northeast. CONCLUSIONS: We identified a vulnerable stroke survivor population with reduced medication access and increased barriers to medical care. Membership in this population has grown substantially from 1997 to 2004, potentially leading to increased recurrent stroke incidence. PMID- 17395862 TI - Serum glial fibrillary acidic protein is related to focal brain injury and outcome after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Aneurysmal subarachnoid hemorrhage (aSAH) stands out from other subtypes of stroke because of the high early mortality and the risk of complications. Serum glial fibrillary acidic protein (s-GFAP) concentrations are increased after stroke. The aim of this study was to investigate whether s-GFAP could be used as a marker of brain damage and outcome after aSAH. METHODS: Serum samples were obtained on a regular basis from 116 adults during a 2-week period after aSAH and analyzed using an enzyme-linked immunosorbent assay. The World Federation of Neurological Surgeons scale was used for neurological evaluation. Outcome was assessed after 1 year and categorized according to the Extended Glasgow Outcome Scale. RESULTS: Increased s-GFAP levels were seen in 81 of the 116 patients. Maximum s-GFAP correlated with World Federation of Neurological Surgeons scale on arrival and on days 10 to 15 (r=0.37, P<0.001 and r=0.47, P<0.001, respectively). Furthermore, maximum s-GFAP levels were increased in the patient group with radiological signs of focal lesions acute or at 1 year, compared with the group without focal lesions (P<0.001 in both comparisons). Patients with secondary events (re-bleeding or ischemia) reached maximum levels later in the series and both maximum and final s-GFAP levels increased compared with the levels in patients without secondary events (P<0.001 in all 3 comparisons). Finally, maximum s-GFAP correlated with outcome (r=-0.48, P<0.001) and s-GFAP was an independent predictor of dichotomized outcome. CONCLUSIONS: s GFAP provides information about brain injury severity and outcome after aSAH, which can be useful as a complement to clinical data. PMID- 17395863 TI - Radiological findings, clinical course, and outcome in asymptomatic moyamoya disease: results of multicenter survey in Japan. AB - BACKGROUND AND PURPOSE: Although the development of a noninvasive MR examination has increased the opportunity to identify asymptomatic patients with moyamoya disease who have experienced no stroke episodes, their clinical features are still unclear. This was the first multicenter, nation-wide survey focused on asymptomatic moyamoya disease in Japan and was designed to clarify their clinical features. METHODS: A clinical database of asymptomatic patients with moyamoya disease was collected from 12 participating hospitals in Japan between 2003 and 2006. In total, 40 patients were enrolled in this historical prospective cohort study. Of these, 6 underwent surgical revascularization, including superficial temporal artery to middle cerebral artery anastomosis and/or pial synangiosis. Their demographic and radiological findings as well as outcome were evaluated. RESULTS: On initial evaluation, cerebral infarction and disturbed cerebral hemodynamics were detected in approximately 20% and 40% of the involved hemispheres, respectively. Angiographical stage was more advanced in more elderly patients. Of 34 nonsurgically treated patients, 7 experienced transient ischemic attack (n=3), ischemic stroke (n=1), or intracranial bleeding (n=3) during follow up periods (mean, 43.7 months). The annual risk for any stroke was 3.2%. Disease progression was associated with ischemic events or silent infarction in 4 of 5 patients. No cerebrovascular event occurred in the 6 patients who underwent surgical revascularization. CONCLUSIONS: The findings revealed that asymptomatic moyamoya disease is not a silent disorder and may potentially cause ischemic or hemorrhagic stroke. Asymptomatic patients with moyamoya disease should be carefully followed-up to further clarify their outcome and to establish the management guideline for them. PMID- 17395864 TI - A novel, self-expanding, nitinol stent in medically refractory intracranial atherosclerotic stenoses: the Wingspan study. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the safety and performance of the Wingspan stent system and Gateway percutaneous transluminal angioplasty balloon catheter in the treatment of high-grade, intracranial atherosclerotic lesions in patients who had failed medical therapy. METHODS: In this prospective, multicenter, single-arm study, medically refractory patients with a modified Rankin score < or =3 and recurrent symptoms attributable to angiographically demonstrated intracranial stenosis > or =50% in a vessel 2.5 to 4.5 mm in diameter were enrolled. Intracranial lesions were predilated with an undersized Gateway balloon catheter to 80% of the native vessel diameter, followed by deployment of the self-expanding Wingspan stent to facilitate further remodeling of the atherosclerotic plaque and to maintain vessel patency. Neurologic examinations and angiograms were performed at 6 months after the procedure. RESULTS: Among the 45 patients enrolled, the degree of stenosis was reduced from a baseline of 74.9+/-9.8% to 31.9+/-13.6% after stenting and 28+/ 23.2% at the 6-month follow-up. The 30-day composite ipsilateral stroke/death rate was 4.5% (2/44); at the 6-month follow-up, the ipsilateral stroke/death rate was 7.0%, the rate for all strokes was 9.7%, and all-cause mortality was 2.3%. Physician-reported follow-up in 43 patients (average of 13 months) conducted outside the study protocol (not adjudicated by the clinical event committee) reported 1 additional ipsilateral stroke. CONCLUSIONS: In medically refractory patients with high-grade intracranial atherosclerotic stenoses, a new treatment paradigm involving predilation with an undersized Gateway percutaneous transluminal angioplasty balloon catheter and placement of a self-expanding Wingspan stent system appears to be safe, may facilitate remodeling, and may contribute to favorable angiographic outcomes. PMID- 17395865 TI - Atrial fibrillation, stroke, and cognition: a longitudinal population-based study of people aged 85 and older. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate the association between atrial fibrillation (AF), stroke, dementia, and their correlation with brain pathology in subjects aged 85 years or older. METHODS: This is a prospective 9-year follow-up population based study in Vantaa, a town in Southern Finland; 553 subjects (92% of the total population) aged 85 years or older were clinically examined by a neurologist. The presence of AF was collected from the medical records or examined by ECG or ambulatory ECG. Neuropathological examination was conducted in more than half of the clinically examined subjects. RESULTS: AF was significantly associated with stroke at baseline; 32% of patients with AF had clinical evidence of stroke compared with 16.7% of those without such evidence (P<0.001). Dementia at baseline was significantly associated with age, clinical stroke, and the presence of apolipoprotein E epsilon4 allele, but not with sex, education, or vascular risk factors. Multiple regression analysis including neuropathological results showed that dementia was significantly associated with education (OR, 0.89; 95% CI, 0.80 to 0.98; P=0.019), the beta amyloid load in the brain (OR, 1.26; 95% CI, 1.13 to 1.39; P<0.001) and with the vascular pathology (OR, 2.03; 95% CI, 1.14 to 3.62; P=0.016), but not with sex, age at death, apolipoprotein E epsilon4 allele, or vascular risk factors. CONCLUSIONS: AF is a significant and preventable risk factor for stroke but not for dementia in the very old. The etiology of dementia syndrome in the very old is multifactorial. Both Alzheimer disease pathology and vascular pathology, particularly multiple small infarcts, contribute to cognitive decline. PMID- 17395866 TI - Peripheral inflammation exacerbates damage after global ischemia independently of temperature and acute brain inflammation. AB - BACKGROUND AND PURPOSE: Concomitant infection can exacerbate damage caused by cerebral ischemia. However, the interaction between and relative importance of the febrile and inflammatory components of the immune response is still unknown. METHODS: Male Sprague-Dawley rats were subjected to a 2-vessel occlusion with hypotension, immediately followed by intraperitoneal injection of lipopolysaccharide or pyrogen-free saline. RESULTS: Inflammation immediately after 2-vessel occlusion exacerbated hippocampal cell loss at 3 days and enhanced anxiety-related behaviors in the elevated plus maze and open field. These effects were not associated with differences in body temperature changes or with hippocampal pro-inflammatory cytokine production or hippocampal microglial activation. CONCLUSIONS: We show a previously undocumented dissociation between lipopolysaccharide-exacerbated damage after global ischemia in the rat and the temperature and acute brain immune response, indicating that the mechanism for enhanced lipopolysaccharide damage is hippocampal cytokine and temperature independent in this case. PMID- 17395867 TI - Association between physician volume and hospitalization costs for patients with stroke in Taiwan: a nationwide population-based study. AB - BACKGROUND AND PURPOSE: Past studies consistently show an inverse relationship between physicians' surgical procedures/diagnoses volume and cost. There is little information available on this aspect of stroke care. We used nationwide population-based data to explore the association between physician case volume and costs per discharge for patients with stroke. METHODS: Data on all 83,748 hospitalizations for stroke in 2004, treated by 3757 physicians in Taiwan, from Taiwan's National Health Insurance Research Database, was analyzed using hierarchical linear regression modeling to explore associations between costs per discharge and physician case volume (one to 44 cases=low volume, 44 to 135=medium volume, > or =136 cases=high volume) adjusting for patient's age, gender, comorbidities, and stroke type; hospital ownership, teaching status, and geographic region; and physician demographics. RESULTS: Unadjusted mean cost per discharge was highest for patients treated by low-volume physicians, at NT $79,993 compared with NT $78,588 for medium-volume physicians and NT $43,942 for high-volume physicians (P<0.001). Adjusted for patient, hospital, and physician variables, low-volume physicians had a mean case cost of NT $27,729 higher than high-volume physicians (P=0.001) and NT $7761 higher than medium-volume physicians (P=0.027). CONCLUSIONS: Our data confirm an inverse volume-cost relationship for stroke care in Taiwan. After adjusting for patient, hospital, and physician characteristics, the potential cost savings if all patients were treated or supervised by high-volume physicians could be 41.0% of the mean treatment cost incurred by low-volume physicians. PMID- 17395868 TI - A single infusion of zoledronate prevents bone loss after stroke. AB - BACKGROUND AND PURPOSE: Stroke is a major risk factor for hip fracture. Patients with intermediate rather than severe or mild stroke deficits at the time of hospital discharge have the most fractures. This proof-of-concept study evaluated the efficacy of a single infusion of zoledronate, an intravenous bisphosphonate, in preserving hip bone density after stroke. METHODS: In a 1-year randomized, double-blind, placebo-controlled, clinical trial, 27 newly hemiplegic patients (6 females, 21 males) with acute stroke were assigned to receive 4 mg of the intravenous zoledronate (n=14) or placebo (n=13) within 35 days. Strict inclusion criteria were followed-up to ensure recruited patients were likely to have residual functional impairment. Both groups received calcium and vitamin D supplementation. The primary outcome measure was the change in bone mineral density (BMD; Lunar Prodigy) at the hemiplegic hip during the year of investigation. RESULTS: The treatment was generally well tolerated. Mean total hip BMD was unchanged in the hemiplegic hip of the zoledronate group (mean 0.0% change), whereas in the placebo group the total hip BMD changed by -5.5%, with the greatest bone loss observed in the trochanteric subregion (mean, -8.1%). On the unaffected side the mean change in total hip BMD was +1.0% with zoledronate versus a mean change of -2.7% without. Repeated measures ANOVA confirmed the significance of the differences between groups at both hips (hemiplegic, P<0.001; unaffected, P=0.002). CONCLUSIONS: Stroke patients were protected from the deleterious effects of hemiplegia on hip bone density for at least 1 year after a single infusion of zoledronate. PMID- 17395869 TI - Long-term outcome after angioplasty and stenting for symptomatic vertebral artery stenosis compared with medical treatment in the Carotid And Vertebral Artery Transluminal Angioplasty Study (CAVATAS): a randomized trial. AB - BACKGROUND AND PURPOSE: The long-term outcome of endovascular intervention compared with best medical management of patients with symptomatic vertebral artery stenosis is uncertain. We therefore compared these treatments in a randomized trial with long-term follow-up. METHODS: In the international, multicenter Carotid And Vertebral Artery Transluminal Angioplasty Study, 16 patients with symptomatic vertebral artery stenosis were randomized in equal proportions to receive endovascular therapy (balloon angioplasty or stenting) or best medical treatment alone. An independent neurologist followed up the patients for as long as 8 years. RESULTS: Endovascular intervention was technically successful in all 8 patients, but 2 patients experienced transient ischemic attack at the time of endovascular treatment. There were no deaths or strokes in any arterial territory within the first 30 days. During a mean follow-up period of 4.7 years, no patient in either treatment group experienced a vertebrobasilar territory stroke, but 3 patients in each treatment arm died of myocardial infarction or carotid territory stroke, and 1 endovascular patient had a nonfatal carotid territory stroke. CONCLUSIONS: Patients with vertebral artery stenosis were more likely to have carotid territory stroke and myocardial infarction during follow-up than have recurrent vertebrobasilar stroke. The trial failed to show a benefit of endovascular treatment of vertebral artery stenosis, but the numbers of patients included was small. Larger randomized trials are required to determine whether vertebral artery stenting is justified in patients at higher risk of vertebrobasilar stroke. Treatment of patients with vertebral artery stenosis should focus on global reduction of vascular risk, including prevention of carotid territory stroke and myocardial infarction. PMID- 17395870 TI - Retreatment of ruptured cerebral aneurysms in patients randomized by coiling or clipping in the International Subarachnoid Aneurysm Trial (ISAT). AB - BACKGROUND AND PURPOSE: Because the long-term security of endovascular treatments remains uncertain, a follow-up study of the patients treated in the International Subarachnoid Aneurysm Trial was performed to compare the frequency, timing, and consequences of aneurysm recurrence. METHODS: Patient data were reclassified by actual treatment performed. Aneurysm and patient characteristics, including occlusion grades, time and type of retreatment, and clinical outcomes, were compared. The relationship between these variables and late retreatment as a surrogate for recurrence was analyzed by means of the Cox proportional hazards model. RESULTS: Retreatment was performed in 191 of 1096 (17.4%) patients after primary endovascular coiling (EVT) and in 39 of 1012 patients (3.8%) after neurosurgical clipping. After EVT, 97 (8.8%) patients were retreated early and 94 (9.0%) late, 7 (0.6%) after rebleeding and 87 (8.3%) without. The mean time to late retreatment was 20.7 months. After neurosurgical clipping, 30 (2.9%) patients were retreated early and 9 (0.85%) late, 3 (0.3%) after rebleeding and 6 (0.6%) without. The mean time to late retreatment was 5.7 months. The hazard ratio (HR) for retreatment after EVT was 6.9 (95% CI=3.4 to 14.1) after adjustment for age (P=0.001, HR=0.97, 95% CI=0.95 to 0.98), lumen size (P=0.006, HR=1.1, 95% CI=1.03 to 1.18), and incomplete occlusion (P<0.001, HR=7.6, 95% CI=3.3 to 17.5). CONCLUSIONS: Late retreatment was 6.9 times more likely after EVT. Younger age, larger lumen size, and incomplete occlusion were risk factors for late retreatment after EVT. After neurosurgical clipping, retreatments were earlier; whereas EVT retreatments continued to be performed throughout the follow up period. Short-term follow-up imaging is therefore insufficient to detect recurrences after EVT. PMID- 17395871 TI - Clinical impact of NXY-059 demonstrated in the SAINT I trial: derivation of number needed to treat for benefit over entire range of functional disability. AB - BACKGROUND AND PURPOSE: The SAINT I trial demonstrated that the neuroprotective agent NXY-059 improves the distribution of acute stroke patient outcomes on the modified Rankin Scale (mRS) of global disability. Standard dichotomized number needed-to-treat (NNT) analyses for the magnitude of treatment benefit range widely, from 22.7 to infinity, and each capture only a portion of the observed beneficial shift in outcomes. Derivation of an NNT value reflecting the treatment's benefit over the entire range of the mRS is required to describe the clinical import of the trial results. METHODS: The minimum and maximum possible NNTs for benefit over the range of mRS global disability outcomes were calculated by completing a joint distribution outcome table for a model population of 1000 patients, shifting responders by the greatest and smallest possible increments, respectively. The biologically most plausible NNT within this range was derived by having 10 neurologist and emergency-physician acute stroke-care experts independently specify the joint distribution of outcomes in model samples of 1000 patients assigned to placebo and active therapy. RESULTS: The minimum possible NNT for benefit incorporating all mRS state transitions is 7.9 and the maximum is 16.7. The biologically most plausible NNT for 1 additional patient to have a better outcome by 1 or more grades on the mRS outcomes is 9.8 (95% CI, 8.7 to 10.9). CONCLUSIONS: Considering improvements in global disability over the entire outcome range, the SAINT I trial results indicate that for every 100 patients treated, approximately 10 will benefit and none be harmed as a result of treatment. PMID- 17395872 TI - Creation of a biological pacemaker by cell fusion. AB - As an alternative to electronic pacemakers, we explored the feasibility of converting ventricular myocytes into pacemakers by somatic cell fusion. The idea is to create chemically induced fusion between myocytes and syngeneic fibroblasts engineered to express HCN1 pacemaker channels (HCN1-fibroblasts). HCN1 fibroblasts were fused with freshly isolated guinea pig ventricular myocytes using polyethylene-glycol 1500. In vivo fused myocyte-HCN1-fibroblast cells exhibited spontaneously oscillating action potentials; the firing frequency increased with beta-adrenergic stimulation. The heterokaryons created ectopic ventricular pacemaker activity in vivo at the site of cell injection. Coculture of nonfused HCN1-fibroblasts and myocytes without polyethylene-glycol 1500 revealed no evidence of dye transfer, demonstrating that the I(f)-mediated pacemaker activity arises from heterokaryons rather than electrotonic coupling. This nonviral, non-stem cell approach enables autologous, adult somatic cell therapy to create biopacemakers. PMID- 17395873 TI - Endothelial progenitor cells stimulate cerebrovascular production of prostacyclin by paracrine activation of cyclooxygenase-2. AB - In the present study we hypothesized that endothelial progenitor cells (EPCs) enhance production of vasoprotective substances in cerebral arteries. Isolated mononuclear cells from rabbit peripheral blood were cultured in endothelial growth medium (EGM-2) for 7 days to yield EPCs. Rabbit basilar arteries were exposed to autologous EPCs ( approximately 5x10(5) cells) in vitro or in vivo. Twenty-four hours after intracisternal delivery of autologous EPCs, basilar arteries were isolated and expression of vasoregulatory proteins, production of prostacyclin (PGI(2)), and cAMP were determined. Arteries transplanted with EPCs demonstrated increased protein expression of cyclooxygenase-2 and PGI(2) in adventitia, media, and endothelium. Furthermore, production of PGI(2) and arterial content of cAMP, second messenger for PGI(2), were significantly augmented after transplantation of EPCs. In contrast, production of thromboxane A(2) was significantly reduced, whereas production of prostaglandin E(2) remained unchanged. The increased production of PGI(2) and arterial content of cAMP were inhibited only by a selective cyclooxygenase-2 inhibitor, NS-398. In vitro or in vivo treatment of basilar artery with conditioned media from EPCs also caused increase in cyclooxygenase-2 and PGI(2) synthase protein expression associated with elevation of cAMP. Our results suggest that in cerebral arteries, paracrine effect of EPCs promotes vasoprotection by increasing PGI(2) production and intracellular concentration of cAMP. This effect appears to be mediated by activation of arachidonic acid metabolism via stimulation of cyclooxygenase 2/PGI(2) synthase pathway. PMID- 17395874 TI - Inhibition of endogenous Mst1 prevents apoptosis and cardiac dysfunction without affecting cardiac hypertrophy after myocardial infarction. AB - Mammalian sterile 20-like kinase-1 (Mst1) plays an important role in mediating cardiac myocyte apoptosis in response to ischemia/reperfusion. Whether or not Mst1 is also involved in the long-term development of heart failure after myocardial infarction (MI) is unknown. We addressed this issue using transgenic mice with cardiac specific overexpression of dominant negative Mst1 (Tg-DN-Mst1). The left coronary artery was permanently ligated, and the size of MI was similar between Tg-DN-Mst1 and nontransgenic controls (NTg). After 4 weeks, Mst1 was significantly activated in the remodeling area in NTg, but not in Tg-DN-Mst1. Although left ventricular (LV) enlargement was significantly attenuated in Tg-DN Mst1 compared with NTg, neither LV weight/body weight nor myocyte cross sectional area was statistically different between Tg-DN-Mst1 and NTg. LV ejection fraction was significantly greater in Tg-DN-Mst1 than in NTg (53 versus 38%, P<0.01), whereas LV end-diastolic pressure (6 versus 12 mm Hg, P<0.05) and lung weight/body weight (9.8 versus 12.2 P<0.05) were significantly smaller in Tg-DN Mst1 than in NTg. The number of TUNEL-positive myocytes (0.17 versus 0.28%, P<0.05) and amount of interstitial fibrosis (5.0 versus 7.1%, P<0.05) in the remodeling area were significantly less in Tg-DN-Mst1 than in NTg. Upregulation of matrix metalloproteinase 2 and proinflammatory cytokines was significantly attenuated in Tg-DN-Mst1. These results indicate that endogenous Mst1 plays an important role in mediating cardiac dilation, apoptosis, fibrosis, and cardiac dysfunction, but not cardiac hypertrophy, after MI. Inhibition of Mst1 improves cardiac function without attenuating cardiac hypertrophy. Thus, Mst1 may be an important target of heart failure treatment. PMID- 17395875 TI - The BTB-Kelch protein KLEIP controls endothelial migration and sprouting angiogenesis. AB - Sprouting and invasive migration of endothelial cells are important steps of the angiogenic cascade. Vascular endothelial growth factor (VEGF) induces angiogenesis by activating intracellular signal transduction cascades, which regulate endothelial cell morphology and function. BTB-kelch proteins are intracellular proteins that control cellular architecture and cellular functions. The BTB-kelch protein KLEIP has been characterized as an actin-binding protein that interacts with the nucleotide exchange factor ECT2. We report that KLEIP is preferentially expressed in endothelial cells, suggesting that it may play a critical role in controlling the functions of migrating, proliferating, and invading endothelial cells during angiogenesis. KLEIP mRNA level in endothelial cells is strongly regulated by hypoxia which is controlled by hypoxia-inducible factor-1alpha. Functional analysis of KLEIP in endothelial cells revealed that it acts as an essential downstream regulator of VEGF- and basic fibroblast growth factor-induced migration and in-gel sprouting angiogenesis. Yet, it is not involved in controlling VEGF- or basic fibroblast growth factor-mediated proliferative responses. The depletion of KLEIP in endothelial cells blunted the VEGF-induced activation of the monomeric GTPase RhoA but did not alter the VEGF stimulated activation of extracellular signal-regulated kinase 1/2. Moreover, VEGF induced a physical association of KLEIP with the guanine nucleotide-exchange factor ECT2, the depletion of which also blunted VEGF-induced sprouting. We conclude that the BTB-kelch protein KLEIP is a novel regulator of endothelial function during angiogenesis that controls the VEGF-induced activation of Rho GTPases. PMID- 17395876 TI - High pressure promotes monocyte adhesion to the vascular wall. AB - Hypertension is a known risk factor for the development of atherosclerosis. To assess how mechanical factors contribute to this process, mouse carotid arteries were maintained in organ culture at normal (80 mm Hg) or high (150 mm Hg) intraluminal pressure for 1, 6, 12, or 24 hours. Thereafter, fluorescent human monocytic cells (U937) were injected intraluminally and allowed to adhere for 30 minutes before washout. U937 adhesion was increased in vessels kept at 150 mm Hg 12 hours (23.5+/-5.7 versus 9.9+/-2.2 cells/mm at 80 mm Hg; P<0.05) or 24 hours (26.7+/-5.7 versus 8.8+/-1.5 cells/mm; P<0.05). At 24 hours, high pressure was associated with increased mRNA expression of monocyte chemoattractant protein-1, interleukin-6, keratinocyte-derived chemokine, and vascular cell adhesion molecule-1 (6.9+/-2.1, 4.4+/-0.1, 9.8+/-2.8, and 2.4+/-0.1-fold respectively; P<0.05), as assessed by quantitative RT-PCR and corroborated by immunohistochemistry, which also revealed an increase in intracellular adhesion molecule-1 expression. Nuclear factor kappaB inhibition using SN50 peptide abolished the overexpression of chemokines and adhesion molecules and reduced U937 adhesion in vessels at 150 mm Hg. Moreover, treatment of vessels and cells with specific neutralizing antibodies established that monocyte chemoattractant protein-1, interleukin-6, and keratinocyte-derived chemokine released from vessels at 150 mm Hg primed the monocytes, increasing their adhesion to vascular cell adhesion molecule-1 but not intracellular adhesion molecule-1 via alpha4beta1 integrins. The additive effect of chemokines on the adhesion of U937 cells to vascular cell adhesion molecule-1 was confirmed by in vitro assay. Finally, pressure-dependent U937 adhesion was blunted in arteries from mice overexpressing endothelial NO synthase. Hence, high intraluminal pressure induces cytokine and adhesion molecule expression via nuclear factor kappaB, leading to monocytic cell adhesion. These results indicate that hypertension may directly contribute to the development of atherosclerosis through nuclear factor kappaB induction. PMID- 17395877 TI - The matrix revolutions: matrix metalloproteinase, vasculogenesis, and ischemic tissue repair. PMID- 17395878 TI - Selective depletion of macrophages in atherosclerotic plaques: myth, hype, or reality? PMID- 17395879 TI - Physiological smooth muscle cell apoptosis contributes to the uterine vascular remodeling in human early pregnancy. PMID- 17395880 TI - Move on!: smooth muscle cell motility paired down. PMID- 17395881 TI - One gene, many proteins: alternative splicing of the ryanodine receptor gene adds novel functions to an already complex channel protein. PMID- 17395882 TI - Hey, there's a hole in my heart. PMID- 17395883 TI - Do glucose and lipids exert independent effects on atherosclerotic lesion initiation or progression to advanced plaques? AB - It is becoming increasingly clear that suboptimal blood glucose control results in adverse effects on large blood vessels, thereby accelerating atherosclerosis and cardiovascular disease, manifested as myocardial infarction, stroke, and peripheral vascular disease. Cardiovascular disease is accelerated by both type 1 and type 2 diabetes. In type 1 diabetes, hyperglycemia generally occurs in the absence of elevated blood lipid levels, whereas type 2 diabetes is frequently associated with dyslipidemia. In this review article, we discuss hyperglycemia versus hyperlipidemia as culprits in diabetes-accelerated atherosclerosis and cardiovascular disease, with emphasis on studies in mouse models and isolated vascular cells. Recent studies on LDL receptor-deficient mice that are hyperglycemic, but exhibit no marked dyslipidemia compared with nondiabetic controls, show that diabetes in the absence of diabetes-induced hyperlipidemia is associated with an accelerated formation of atherosclerotic lesions, similar to what is seen in fat-fed nondiabetic mice. These effects of diabetes are masked in severely dyslipidemic mice, suggesting that the effects of glucose and lipids on lesion initiation might be mediated by similar mechanisms. Recent evidence from isolated endothelial cells demonstrates that glucose and lipids can induce endothelial dysfunction through similar intracellular mechanisms. Analogous effects of glucose and lipids are also seen in macrophages. Furthermore, glucose exerts many of its cellular effects through lipid mediators. We propose that diabetes without associated dyslipidemia accelerates atherosclerosis by mechanisms that can also be activated by hyperlipidemia. PMID- 17395884 TI - Endothelial cell migration during angiogenesis. AB - Endothelial cell migration is essential to angiogenesis. This motile process is directionally regulated by chemotactic, haptotactic, and mechanotactic stimuli and further involves degradation of the extracellular matrix to enable progression of the migrating cells. It requires the activation of several signaling pathways that converge on cytoskeletal remodeling. Then, it follows a series of events in which the endothelial cells extend, contract, and throw their rear toward the front and progress forward. The aim of this review is to give an integrative view of the signaling mechanisms that govern endothelial cell migration in the context of angiogenesis. PMID- 17395885 TI - Defective mitochondrial biogenesis: a hallmark of the high cardiovascular risk in the metabolic syndrome? AB - The metabolic syndrome is a group of risk factors of metabolic origin that are accompanied by increased risk for type 2 diabetes mellitus and cardiovascular disease. These risk factors include atherogenic dyslipidemia, elevated blood pressure and plasma glucose, and a prothrombotic and proinflammatory state. The condition is progressive and is exacerbated by physical inactivity, advancing age, hormonal imbalance, and genetic predisposition. The metabolic syndrome is a particularly challenging clinical condition because its complex molecular basis is still largely undefined. Impaired cell metabolism has, however, been suggested as a relevant pathophysiological process underlying several clinical features of the syndrome. In particular, defective oxidative metabolism seems to be involved in visceral fat gain and in the development of insulin resistance in skeletal muscle. This suggests that mitochondrial function may be impaired in the metabolic syndrome and, thus, in the consequent cardiovascular disease. We have recently found that mitochondrial biogenesis and function are enhanced by nitric oxide in various cell types and tissues, including cardiac muscle. Increasing evidence suggests that this mediator acts as a metabolic sensor in cardiomyocytes. This implies that a defective production of nitric oxide might be linked to dysfunction of the cardiomyocyte metabolism. Here we summarize some recent findings and propose a hypothesis for the high cardiovascular risk linked to the metabolic syndrome. PMID- 17395886 TI - Ang-2 and PDGF-BB cooperatively stimulate human peripheral blood monocyte fibrinolysis. AB - Angiopoietin-2 (Ang-2) is a growth factor, which was identified originally as playing a critical role in vessel remodeling during angiogenesis. More recent evidence has indicated additional involvement in vascular homeostatic responses such as coagulation and inflammation, which are central to wound healing. We therefore determined whether a relationship existed between Ang-2 and monocytes, one of the initial cell types to be recruited to a wound, in the context of fibrin clot invasion. Ang-2 significantly increased monocyte invasion of fibrin in the presence of serum. In the absence of serum, it required a combination of Ang-2 and platelet-derived growth factor BB (PDGF-BB) to increase invasion by threefold. Furthermore, it was shown that the heightened invasion was dependent on serine proteases and matrix metalloproteinases (MMPs) and that the combination of Ang-2 and PDGF-BB increased urokinase plasminogen-activator receptor expression, as well as MMP-9 and membrane type 1 MMP expression. These data give further credence to the concept of Ang-2 as a key regulator of several essential phases of wound healing. PMID- 17395887 TI - COX-2 expression induced by diesel particles involves chromatin modification and degradation of HDAC1. AB - Cyclooxygenase-2 (COX-2) plays an important role in the inflammatory response induced by physiologic and stress stimuli. Exposure to diesel exhaust particulate matter (DEP) has been shown to induce pulmonary inflammation and exacerbate asthma and chronic obstructive pulmonary disease. DEP is a potent inducer of inflammatory reponses in human airway epithelial cells. The mechanism through which DEP inhalation induces inflammatory mediator expression is not understood. In this report, we demonstrate that DEP can induce the expression of COX-2 gene in a human bronchial epithelial cell line (BEAS-2B) at both transcriptional and protein levels. The induction of COX-2 gene expression involves chromatin modification, in particular acetylation and deacetylation of histones. We show that exposure to DEP increases the acetylation of histone H4 associated with the COX-2 promoter and causes degradation of histone deacetylase 1 (HDAC1). Further, we establish that HDAC1 plays a pivotal role in mediating the transcriptional activation of the COX-2 gene in BEAS-2B cells exposed to DEP, supported by evidence that the down-regulation of HDAC1 using siRNA leads to activation of COX 2 gene expression, whereas overexpression of HDAC1 results in its repression. Finally, DEP exposure induced recruitment of histone acetyltransferase (HAT) p300 to the promoter of the COX-2 gene, suggesting that acetylation is also important in regulating its expression in response to DEP exposure. These results show for the first time acetylation via selective degradation of HDAC1, and that recruitment of HAT plays an important role in DEP-induced expression of the COX-2 gene. PMID- 17395889 TI - Isolation of an adult mouse lung mesenchymal progenitor cell population. AB - Contained within the adult lung are differentiated mesenchymal cell types (cartilage, smooth muscle, and myofibrobasts) that provide structural support for airways and vessels. Alterations in the number and phenotype of these cells figure prominently in the pathogenesis of a variety of lung diseases. While these cells are thought to arise locally, progenitors have yet to be purified. In previous work, we developed a method for isolating progenitors from lung tissue: this technique takes advantage of the unique ability of cell populations enriched for somatic stem and progenitor activity to efflux the vital dye Hoechst 33342, a feature that permits isolation by flow cytometry-based procedures. Using this method, we determined that a rare population of mesenchymal progenitors resides within the CD45- CD31- Hoechst low fraction of the adult murine lung. Similar to other mesenchymal progenitors, these cells express Sca-1, CD106, and CD44; can be serially passaged; and can differentiate to smooth muscle, cartilage, bone, and fat. Overall, these findings demonstrate that a phenotypically distinct mesenchymal progenitor resides within the adult murine lung, and provide a scheme for their isolation and study. PMID- 17395888 TI - Apical oxidative hyaluronan degradation stimulates airway ciliary beating via RHAMM and RON. AB - Hyaluronan (HA) is synthesized in high-molecular-weight form at the apical pole of airway epithelial cells, covering the luminal surface. When human airway epithelial cells grown and redifferentiated at the air-liquid interface (ALI) were exposed to xanthine/xanthine oxidase (X/XO), ciliary beat frequency (CBF) increased. This effect was blocked by superoxide dismutase (SOD) and catalase. Inhibition of hyaluronan synthesis inhibited the CBF response to X/XO, while addition of exogenous HA amplified it. A functionally blocking antibody to the receptor for hyaluronic acid-mediated motility (RHAMM) reduced the CBF response to X/XO. Since RHAMM has no transmembrane domain and thus cannot signal on its own, the association of RHAMM with recepteur d'origine nantais (RON), a member of the hepatocyte growth factor receptor family, was explored. Immunohistochemistry of human airway epithelium showed co-localization of RHAMM and RON at the apex of ciliated cells. Physical association of RHAMM and RON was confirmed with co immunoprecipitations. Macrophage-stimulating protein (MSP), an agonist of RON, stimulated CBF. Genistein, a nonspecific tyrosine kinase inhibitor, and MSP beta chain (beta-MSP), a specific RON inhibitor, blocked the X/XO-induced CBF increase. HA present in the apical secretions of human airway epithelial cells was shown to degrade upon exposure to X/XO, a process inhibited by SOD. Low molecular-weight HA fragments stimulated CBF, an effect blocked by anti-RHAMM antibody and genistein. These data suggest that high molecular form HA is broken down by reactive oxygen species to form low-molecular-weight fragments that signal via RHAMM and RON to stimulate CBF. PMID- 17395890 TI - Alpha1-antitrypsin suppresses TNF-alpha and MMP-12 production by cigarette smoke stimulated macrophages. AB - We have previously observed that mice exposed to cigarette smoke and treated with exogenous alpha(1)-antitrypsin (A1AT) were protected against the development of emphysema and against smoke-induced increases in serum TNF-alpha. To investigate possible mechanisms behind this latter observation, we cultured alveolar macrophages lavaged from C57 mice. Smoke-conditioned medium caused alveolar macrophages to increase secretion of macrophage metalloelastase (MMP-12) and TNF alpha, and this effect was suppressed in a dose-response fashion by addition of A1AT. Macrophages from animals exposed to smoke in vivo and then lavaged also failed to increase MMP-12 and TNF-alpha secretion when the animals were pretreated with A1AT. Because proteinase activated receptor-1 (PAR-1) is known to control MMP-12 release, macrophages were treated with the G protein-coupled receptor inhibitor, pertussis toxin; this suppressed both TNF-alpha and MMP-12 release, while a PAR-1 agonist (TRAP) increased TNF-alpha and MMP-12 release. Smoke-conditioned medium caused increased release of the prothrombin activator, tissue factor, from macrophages. Hirudin, a thrombin inhibitor, and aprotinin, an inhibitor of plasmin, reduced smoke-mediated TNF-alpha and MMP-12 release, and A1AT inhibited both plasmin and thrombin activity in a cell-free functional assay. These findings extend our previous suggestion that TNF-alpha production by alveolar macrophages is related to MMP-12 secretion. They also suggest that A1AT can inhibit thrombin and plasmin in blood constituents that leak into the lung after smoke exposure, thereby preventing PAR-1 activation and MMP-12/TNF-alpha release, and decreasing smoke-mediated inflammatory cell influx. PMID- 17395891 TI - Attenuation of vascular permeability by methylnaltrexone: role of mOP-R and S1P3 transactivation. AB - Endothelial cell (EC) barrier dysfunction (i.e., increased vascular permeability) is observed in inflammatory states, tumor angiogenesis, atherosclerosis, and both sepsis and acute lung injury. Therefore, agents that preserve vascular integrity have important clinical therapeutic implications. We examined the effects of methylnaltrexone (MNTX), a mu opioid receptor (mOP-R) antagonist, on human pulmonary EC barrier disruption produced by edemagenic agents including morphine, the endogenous mOP-R agonist DAMGO, thrombin, and LPS. Pretreatment of EC with MNTX (0.1 muM, 1 h) or the uncharged mOP-R antagonist naloxone attenuated morphine- and DAMGO-induced barrier disruption in vitro. However, MNTX, but not naloxone, pretreatment of EC inhibited thrombin- and LPS-induced barrier disruption, indicating potential mOP-R-independent effects of MNTX. In addition, intravenously delivered MNTX attenuated LPS-induced vascular hyperpermeability in the murine lung. We next examined the mechanistic basis for this MNTX barrier protection and observed that silencing of mOP-R attenuated the morphine- and DAMGO-induced EC barrier disruption, but not the permeability response to either thrombin or LPS. Because activation of the sphingosine 1-phosphate receptor, S1P(3), is key to a number of barrier-disruptive responses, we examined the role of this receptor in the permeability response to mOP-R ligation. Morphine, DAMGO, thrombin, and LPS induced RhoA/ROCK-mediated threonine phosphorylation of S1P(3), which was blocked by MNTX, suggesting S1P(3) transactivation. In addition, silencing of S1P(3) receptor expression (siRNA) abolished the permeability response to each edemagenic agonist. These results indicate that MNTX provides barrier protection against edemagenic agonists via inhibition of S1P(3) receptor activation and represents a potentially useful therapeutic agent for syndromes of increased vascular permeability. PMID- 17395892 TI - The effect of omeprazole on the bioavailability and safety of garenoxacin in healthy volunteers. AB - The effect of coadministration of omeprazole on the bioavailability of oral garenoxacin was evaluated in an open-label study in 14 healthy subjects. Single dose pharmacokinetics of garenoxacin were determined with and without steady state omeprazole. Following an oral dose of garenoxacin 600 mg on day 1, serial blood samples were obtained over the next 72 hours. Omeprazole 40 mg once daily was administered from days 4 to 10. A second pharmacokinetic assessment of garenoxacin was conducted on day 8. Geometric means for the maximum observed concentration and area under the concentration-time curve from time 0 extrapolated to infinity were 9.6 microg/mL (18.2%) and 132.0 microg.h/mL (18.9%), respectively, for garenoxacin alone and 9.3 microg/mL (21.6%) and 140.4 microg.h/mL (22.1%), respectively, for coadministered garenoxacin and omeprazole. The 90% confidence interval for the ratio of geometric means (with/without omeprazole) for both variables was contained within 0.80 to 1.25, and the bioavailability of garenoxacin was not affected by the concomitant administration of omeprazole. PMID- 17395893 TI - The pharmacokinetics of taurolidine metabolites in healthy volunteers. AB - Taurolidine is an experimental antibacterial and antiendotoxic compound whose clinical utility as an antitumor agent is being investigated in human clinical trials. Taurolidine in aqueous solution exists in equilibrium with taurultam. Taurultam is subsequently transformed to taurinamide. The pharmacokinetic profiles of these metabolites are not well established. In this study, 18 healthy volunteers were administered 5.0 g of taurolidine in 250 mL of 5% polyvinylpyrrolidone in water over 2, 1, or 0.5 hours by intravenous infusion in a parallel-group design. All subjects noted discomfort at the infusion site, although there were no serious adverse events. t(max) generally occurred at the end of infusion for taurinamide, whereas that of taurultam was reached before completion of infusion. The taurolidine metabolite taurultam demonstrated a shorter half-life and lower systemic exposure than taurinamide. Shortening of infusion duration increased the C(max) and AUC of taurultam. Changes in infusion rate did not substantially change the pharmacokinetic parameters of taurinamide. PMID- 17395894 TI - Single-dose pharmacokinetics of multilayer-release methylphenidate and immediate release methylphenidate in children with attention-deficit/hyperactivity disorder. AB - The objective of this study was to compare the single-dose pharmacokinetics of multilayer-release and immediate-release methylphenidate in children with attention-deficit/hyperactivity disorder. Patients 6- to 12-years-old with a DSM IV diagnosis of attention-deficit/hyperactivity disorder were randomized to receive multilayer-release methylphenidate (qd) or immediate-release methylphenidate (bid) at equivalent doses, with a 14-day washout between treatments. Plasma samples were collected predosing and 1, 2, 3, 4, 5, 6, 8, 10, 12, and 24 hours postdose. Pharmacokinetic analysis was conducted on 14 patients (1 female, 13 male; mean age: 9.6 +/- 2.5 years [range, 6-12]). The mean dose of methylphenidate received by these patients in both phases of the study was 38.6 mg/d (range, 20-80 mg/d). The relative AUC(0-t) and C(max 0-4) ratios for multilayer-release compared with immediate-release methylphenidate were 100.8% and 78.8%, respectively. Multilayer-release methylphenidate produces a biphasic concentration-time profile, with a rapid initial increase in plasma concentration that is maintained throughout the school day. PMID- 17395895 TI - Effects of a cannabinoid receptor agonist on colonic motor and sensory functions in humans: a randomized, placebo-controlled study. AB - Cannabinoid receptors (CBR) are located on cholinergic neurons in the brain stem, stomach, and colon. CBR stimulation inhibits motility in rodents. Effects in humans are unclear. Dronabinol (DRO), a nonselective CBR agonist, inhibits colonic motility and sensation. The aim of this study was to compare effects of DRO and placebo (PLA) on colonic motility and sensation in healthy volunteers. Fifty-two volunteers were randomly assigned (double-blind) to a single dose of 7.5 mg DRO or PLA postoperative with concealed allocation. A balloon-manometric assembly placed into the descending colon allowed assessment of colonic compliance, motility, tone, and sensation before and 1 h after oral ingestion of medication, and during fasting, and for 1 h after 1,000-kcal meal. There was an overall significant increase in colonic compliance (P = 0.045), a borderline effect of relaxation in fasting colonic tone (P = 0.096), inhibition of postprandial colonic tone (P = 0.048), and inhibition of fasting and postprandial phasic pressure (P = 0.008 and 0.030, respectively). While DRO did not significantly alter thresholds for first gas or pain sensation, there was an increase in sensory rating for pain during random phasic distensions at all pressures tested and in both genders (P = 0.024). In conclusion, in humans the nonselective CBR agonist, DRO, relaxes the colon and reduces postprandial colonic motility and tone. Increase in sensation ratings to distension in the presence of relaxation of the colon suggests central modulation of perception. The potential for CBR to modulate colonic motor function in diarrheal disease such as irritable bowel syndrome deserves further study. PMID- 17395896 TI - Interleukin-6 inhibits growth hormone-mediated gene expression in hepatocytes. AB - During systemic inflammation, the liver becomes unresponsive to growth hormone (GH), resulting in decreased plasma insulin-like growth factor-I (IGF-I) with concomitant reductions in lean body mass. Transgenic mice that overexpress IL-6 also demonstrate impaired growth and decreased IGF-I. To determine whether IL-6 directly inhibits GH-inducible gene expression, CWSV-1 hepatocytes were incubated with IL-6 (10 ng/ml), then stimulated with recombinant human GH (500 ng/ml, 18 h). The increase in IGF-I and serine protease inhibitor 2.1 (Spi 2.1) mRNA in GH treated cells was inhibited by treatment with IL-6 for 24 h. To investigate potential mechanisms, we examined the effects of IL-6 on GH receptor (GHR) expression and GH signaling via the JAK/signal transducer and activator of transcription (STAT) and MAP kinase pathways. Incubation of cells with IL-6 (10 ng/ml, 24 h) had no effect on GHR abundance or signaling proteins JAK2, STAT5b, and ERK1/2. Although GH transiently increased (2- to 5-fold) the tyrosine phosphorylation of GHR, JAK2, STAT5b, and ERK1/2, IL-6 did not alter these phosphorylation events. However, nuclear protein from IL-6-treated cells demonstrated reduced STAT5 DNA binding (by EMSA) at 15 min (-20%) and 60 min ( 43%) after GH stimulation. To determine whether IL-6 inhibits GH-inducible promoter activity, CWSV-1 cells were transfected with Spi 2.1 or prolactin receptor promoter luciferase vectors, incubated with or without IL-6, then stimulated with GH. The induction of both Spi 2.1 (7.5-fold) and prolactin receptor (4-fold) promoter activity by GH was inhibited by IL-6. In summary, IL-6 mediates hepatic GH resistance by a time-dependent inhibition of GH-inducible promoter activity that is associated with reductions in STAT5 DNA binding. PMID- 17395897 TI - Overexpression of Kruppel-like factor 5 in esophageal epithelia in vivo leads to increased proliferation in basal but not suprabasal cells. AB - Kruppel-like factor 5 (Klf5; also called IKLF or BTEB2), a zinc-finger transcription factor with proproliferative and transforming properties in vitro, is expressed in proliferating cells of gastrointestinal tract epithelia, including in basal cells of the esophagus. Thus, Klf5 is an excellent candidate to regulate esophageal epithelial proliferation in vivo. Nonetheless, the function of Klf5 in esophageal epithelial homeostasis and tumorigenesis in vivo has not previously been determined. Here, we used the ED-L2 promoter of the Epstein-Barr virus to express Klf5 throughout esophageal epithelia. ED-L2/Klf5 transgenic mice were born at the appropriate Mendelian ratio, survived to at least 1 yr of age, and showed no evidence of esophageal dysplasia or cancer. Staining for bromodeoxyuridine (BrdU) demonstrated increased proliferation in the basal layer of ED-L2/Klf5 mice, but no proliferation was seen in suprabasal cells, despite ectopic expression of Klf5 in these cells. Notably, expression of the KLF family member Klf4, which binds the same DNA sequences as Klf5 and which inhibits proliferation and promotes differentiation, was not altered in ED L2/Klf5 transgenic mice. In primary esophageal keratinocytes that overexpressed Klf5, expression of Klf4 still inhibited proliferation and promoted differentiation, providing a possible mechanism for the persistence of keratinocyte differentiation in ED-L2/Klf5 mice. To identify additional targets for Klf5 in esophageal epithelia, we performed functional genomic analyses and identified a total of 15 differentially expressed genes. In summary, while Klf5 positively regulates proliferation in basal cells, it is not sufficient to maintain proliferation in the esophageal epithelium. PMID- 17395898 TI - Enteric neural pathways mediate the anti-inflammatory actions of glucagon-like peptide 2. AB - Glucagon-like peptide-2 (GLP-2) is an important regulator of nutritional absorptive capacity with anti-inflammatory actions. We hypothesized that GLP-2 reduces intestinal mucosal inflammation by activation of vasoactive intestinal polypeptide (VIP) neurons of the submucosal plexus. Ileitis or colitis was induced in rats by injection of trinitrobenzene sulfonic acid (TNBS), or colitis was induced by administration of dextran sodium sulfate (DSS) in drinking water. Subsets of animals received (1-33)-GLP-2 (50 mug/kg sc bid) either immediately or 2 days after the establishment of inflammation and were followed for 3-5 days. The involvement of VIP neurons was assessed by concomitant administration of GLP 2 and the VIP antagonist [Lys(1)-Pro(2,5)-Arg(3,4)-Tyr(6)]VIP and by immunohistochemical labeling of GLP-2-activated neurons. In all models, GLP-2 treatment, whether given immediately or delayed until inflammation was established, resulted in significant improvements in animal weights, mucosal inflammation indices (myeloperoxidase levels, histological mucosal scores), and reduced levels of inflammatory cytokines (IFN-gamma, TNF-alpha, IL-1beta) and inducible nitric oxide synthase, with increased levels of IL-10 in TNBS ileitis and DSS colitis. Reduced rates of crypt cell proliferation and of apoptosis within crypts in inflamed tissues were also noted with GLP-2 treatment. These effects were abolished with coadministration of GLP-2 and the VIP antagonist. GLP 2 was shown to activate neurons and to increase the number of cells expressing VIP in the submucosal plexus of the ileum. These findings suggest that GLP-2 acts as an anti-inflammatory agent through activation of enteric VIP neurons, independent of proliferative effects. They support further studies to examine the role of neural signaling in the regulation of intestinal inflammation. PMID- 17395899 TI - Novel localization of Rab3D in rat intestinal goblet cells and Brunner's gland acinar cells suggests a role in early Golgi trafficking. AB - Rab3D is a small GTP-binding protein that associates with secretory granules of endocrine and exocrine cells. The physiological role of Rab3D remains unclear. While it has initially been implicated in the control of regulated exocytosis, recent deletion-mutation studies have suggested that Rab3D is involved in the biogenesis of secretory granules. Here, we report the unexpected finding that Rab3D also associates with early Golgi compartments in intestinal goblet cells and in Brunner's gland acinar cells. Expression of Rab3D in the intestine was demonstrated by SDS-PAGE and Western blot analysis of homogenates prepared from the rat duodenum and colon. Confocal laser scanning microscopy revealed Rab3D immunofluorescence in the Golgi area of goblet cells of the duodenum and colon and in Brunner's gland acinar cells. There was no colocalization between Rab3D and a trans-Golgi network marker, TGN-38. In contrast, Rab3D colocalized partially with a cis-Golgi marker, GM-130, and with a marker of cis-Golgi and coat protein complex I vesicles, beta-COP. Strong colocalization was observed between Rab3D and the lectins Griffonia simplicifolia agglutinin II and soybean agglutinin, which have been described as markers of the medial and cis-Golgi, respectively. Rabphilin, a putative effector of Rab3D, displayed an identical pattern of Golgi localization. Incubation of colon tissue with carbamylcholine or deoxycholate to stimulate exocytosis by goblet cells caused a partial redistribution of Rab3D to the cytoplasm and mucous granule field and a concomitant transformation of the Golgi architecture. Taken together, the present data suggest that Rab3D and rabphilin may regulate the secretory pathway at a much earlier stage than what has hitherto been assumed. PMID- 17395900 TI - Reproducibility of human brain activity evoked by esophageal stimulation using functional magnetic resonance imaging. AB - Functional MRI is a popular tool for investigating central processing of visceral pain in healthy and clinical populations. Despite this, the reproducibility of the neural correlates of visceral sensation by use of functional MRI remains unclear. The aim of the present study was to address this issue. Seven healthy right-handed volunteers participated in the study. Blood oxygen level-dependent contrast images were acquired at 1.5 T while subjects received nonpainful and painful phasic balloon distensions ("on-off" block design, 10 stimuli per "on" period, 0.3 Hz) to the distal esophagus. This procedure was repeated on two further occasions to investigate reproducibility. Painful stimulation resulted in highly reproducible activation over three scanning sessions in the anterior insula, primary somatosensory cortex, and anterior cingulate cortex. A significant decrease in strength of activation occurred from session 1 to session 3 in the anterior cingulate cortex, primary somatosensory cortex, and supplementary motor cortex, which may be explained by an analogous decrease in pain ratings. Nonpainful stimulation activated similar brain regions to painful stimulation, but with greater variability in signal strength and regions of activation between scans. Painful stimulation of the esophagus produces robust activation in many brain regions. A decrease in subjective perception of pain and brain activity from the first to the final scan suggests that serial brain imaging studies may be affected by habituation. These findings indicate that for brain imaging studies that require serial scanning, development of experimental paradigms that control for the effect of habituation is necessary. PMID- 17395902 TI - GERD is associated with shortened telomeres in the squamous epithelium of the distal esophagus. AB - Telomeres are repetitive DNA sequences located at the ends of chromosomes. Telomeres are shortened by repeated cell divisions and by oxidative DNA damage, and cells with critically shortened telomeres cannot divide. We hypothesized that chronic gastroesophageal reflux disease (GERD)-induced injury of the esophageal squamous epithelium results in progressive telomeric shortening that eventually might interfere with mucosal healing. To address our hypothesis, we compared telomere length and telomerase activity in biopsy specimens of esophageal squamous epithelium from GERD patients and control patients. Endoscopic biopsies were taken from the esophageal squamous epithelium of 38 patients with GERD [10 long-segment Barrett's esophagus (LSBE), 15 short-segment (SSBE), 13 GERD without Barrett's esophagus] and 16 control patients without GERD. Telomere length was assessed using the terminal restriction fragment assay, and telomerase activity was studied by the PCR-based telomeric repeat amplification protocol assay. Patients with GERD had significantly shorter telomeres in the distal esophagus than controls [8.3 +/- 0.5 vs. 10.9 +/- 1.5 (SE) Kbp, P = 0.043]. Among the patients with GERD, telomere length in the distal esophagus did not differ significantly in those with and without Barrett's esophagus (LSBE 7.9 +/- 0.8, SSBE 8.6 +/- 0.9, GERD without BE 8.7 +/- 1.0 Kbp). No significant differences in telomerase activity in the distal esophagus were noted between patients with GERD and controls (4.0 +/- 0.39 vs. 5.2 +/- 0.53 RIUs). Telomeres in the squamous epithelium of the distal esophagus of patients who have GERD, with and without Barrett's esophagus, are significantly shorter than those of patients without GERD despite similar levels of telomerase activity. PMID- 17395901 TI - Enteroendocrine cells express functional Toll-like receptors. AB - Intestinal epithelial cells (IECs) provide a physical and immunological barrier against enteric microbial flora. Toll-like receptors (TLRs), through interactions with conserved microbial patterns, activate inflammatory gene expression in cells of the innate immune system. Previous studies of the expression and function of TLRs in IECs have reported varying results. Therefore, TLR expression was characterized in human and murine intestinal sections, and TLR function was tested in an IEC line. TLR1, TLR2, and TLR4 are coexpressed on a subpopulation of human and murine IECs that reside predominantly in the intestinal crypt and belong to the enteroendocrine lineage. An enteroendocrine cell (EEC) line demonstrated a similar expression pattern of TLRs as primary cells. The murine EEC line STC-1 was activated with specific TLR ligands: LPS or synthetic bacterial lipoprotein. In STC-1 cells stimulated with bacterial ligands, NF kappaB and MAPK activation was demonstrated. Furthermore, the expression of TNF and macrophage inhibitory protein-2 were induced. Additionally, bacterial ligands induced the expression of the anti-inflammatory gene transforming growth factor beta. LPS triggered a calcium flux in STC-1 cells, resulting in a rapid increase in CCK secretion. Finally, conditioned media from STC-1 cells inhibited the production of nitric oxide and IL-12 p40 by activated macrophages. In conclusion, human and murine IECs that express TLRs belong to the enteroendocrine lineage. Using a murine EEC model, a broad range of functional effects of TLR activation was demonstrated. This study suggests a potential role for EECs in innate immune responses. PMID- 17395903 TI - HGF ameliorates a high-fat diet-induced fatty liver. AB - Hepatocyte growth factor (HGF) has various effects especially on epithelial cells. However, the precise role of HGF on lipogenesis is still not fully understood. A high-fat diet was administered to HGF transgenic mice and wild-type control mice in vivo. Furthermore, recombinant human HGF (rhHGF) was administered to HepG2 cell line in vitro. We performed an analysis regarding the factors relating to lipid metabolism. An overexpression of HGF dramatically ameliorates a high-fat diet-induced fatty liver. HGF transgenic mice showed an apparently reduced lipid accumulation in the liver. The activation of microsomal triglyceride transfer protein (MTP) and apolipoprotein B (ApoB) accompanying higher triglyceride levels in the serum were found in HGF transgenic mice on a normal diet. Interestingly, this upregulation of the MTP activation became more apparent in the high-fat diet. In addition, the administration of rhHGF stimulated MTP and ApoB expression while reducing reduced the intracellular lipid content in HepG2 cell line. However, this induction of MTP and ApoB by HGF was clearly inhibited by PD98059 (MAPK inhibitor). In conclusion, the data presented in this study indicated that HGF ameliorates a high-fat diet-induced fatty liver via the activation of MTP and ApoB. PMID- 17395904 TI - Anticoagulation for venous thromboembolism. PMID- 17395905 TI - Conduct disorders in children. PMID- 17395906 TI - Acute coronary syndrome. PMID- 17395907 TI - Erythropoiesis stimulating agents. PMID- 17395908 TI - Institutional racism in mental health care. PMID- 17395909 TI - Perinatal death in twins: Absolute risk: better basis for decision making. PMID- 17395910 TI - NICE is encouraging artificial intervention. PMID- 17395911 TI - Oldest old are not just passive recipients of care. PMID- 17395912 TI - Publisher boycott: It's a hatchet job. PMID- 17395913 TI - Publisher boycott: Editorial misses the point. PMID- 17395914 TI - New system threatens UK clinical research. PMID- 17395915 TI - Dying on the acute take can be improved. PMID- 17395916 TI - Asthma in pregnancy: Attacks seem rare in labour. PMID- 17395919 TI - US judge bans use of term "light cigarette" worldwide. PMID- 17395920 TI - NICE faces legal challenge over Alzheimer's drug. PMID- 17395921 TI - New York Times reveals payments to doctors by drug firms. PMID- 17395922 TI - Drug company payments to doctors still hard to access despite disclosure laws. PMID- 17395924 TI - UN calls for tougher rules to prevent sale of children's organs. PMID- 17395925 TI - Tackle substance misuse in young people early, says NICE. PMID- 17395926 TI - NICE recommends women should choose where to give birth. PMID- 17395927 TI - Sexual healing. PMID- 17395928 TI - UK government fails to stem rise in tuberculosis cases. PMID- 17395931 TI - Gynaecologist wins right to challenge fertility authority over raids at his clinics. PMID- 17395932 TI - Patients with tuberculosis in poor countries must be tested for HIV. PMID- 17395933 TI - Global tuberculosis levels plateau while extensively drug resistant strains increase. PMID- 17395934 TI - UK Biobank gets 10% response rate as it starts recruiting volunteers. PMID- 17395943 TI - Genes on ice. PMID- 17395944 TI - Sweetening the pill. PMID- 17395945 TI - Autism research: What makes an expert? PMID- 17395946 TI - Open all hours. PMID- 17395947 TI - Life and death: The growing gap. PMID- 17395948 TI - Towards sustainable clinical trials. PMID- 17395949 TI - Managing the acute psychotic episode. PMID- 17395950 TI - Spurious hyperkalaemia. PMID- 17395956 TI - The effectiveness of a balance training intervention in reducing the incidence of noncontact ankle sprains in high school football players. AB - BACKGROUND: A high body mass index and previous ankle sprains have been shown to increase the risk of sustaining noncontact inversion ankle sprains in high school football players. HYPOTHESIS: Stability pad balance training reduces the incidence of noncontact inversion ankle sprains in football players with increased risk. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Height, body mass, history of previous ankle sprains, and current ankle brace/tape use were documented at the beginning of preseason training in 2 high school varsity football teams for 3 consecutive years (175 player-seasons). Players were categorized as minimal risk, low risk, moderate risk, and high risk based on the history of previous ankle sprain and body mass index. Players in the low-, moderate-, and high-risk groups (ie, any player with a high body mass index and/or a previous ankle sprain) were placed on a balance training intervention on a foam stability pad. Players balanced for 5 minutes on each leg, 5 days per week, for 4 weeks in preseason and twice per week during the season. Postintervention injury incidence was compared with preintervention incidence (107 players-seasons) for players with increased risk. RESULTS: Injury incidence for players with increased risk was 2.2 injuries per 1000 exposures (95% confidence interval, 1.1-3.8) before the intervention and 0.5 (95% confidence interval, 0.2-1.3) after the intervention (P < .01). This represents a 77% reduction in injury incidence (95% confidence interval, 31%-92%). CONCLUSION: The increased risk of a noncontact inversion ankle sprain associated with a high body mass index and a previous ankle sprain was eliminated by the balance training intervention. PMID- 17395957 TI - Failure strengths in distal biceps tendon repair. AB - BACKGROUND: Surgical repair is the most favored treatment for a rupture of the distal biceps tendon. A variety of techniques have been described for distal biceps tendon reattachment, including transosseous sutures, suture anchors, interference screws, and an EndoButton-based technique. HYPOTHESIS: EndoButton and suture anchor have initially stronger fixation strengths than do transosseous sutures, allowing early postoperative rehabilitation. STUDY DESIGN: Controlled laboratory study. METHODS: Single loads to failure and mode of failure of 13 different fixation techniques were determined using 130 human cadaveric elbows. Quantitative computer tomography was performed to exclude differences in bone mineral density as an affecting factor. Repeated-measures analysis of variance was used to assess differences in failure load between repair techniques. RESULTS: The EndoButton-based technique showed a significantly higher failure load (259 +/- 28 N) than did all other techniques (P < .05). No significant differences were seen between the transosseous suture technique (210 +/- 29 N) and most other techniques (P > .05). Failure loads of the TwinFix-QuickT (57 +/- 29 N) and Biocuff screw (105 +/- 28 N) were significantly lower than those of all other repairs (P < .05). CONCLUSION: Significant differences exist in failure loads and modes of failure for the different repair techniques after rupture of the distal biceps tendon. CLINICAL RELEVANCE: The transosseous technique is still a sufficient and cost-saving procedure for repair of the distal biceps tendon. TwinFix-QuickT 5.0 mm and Biocuff screw 5.7 mm had significantly lower failure loads, which might affect early rehabilitation, particularly in patients with poor bone quality. PMID- 17395958 TI - The use of intra-articular hylan G-F 20 in the treatment of symptomatic osteoarthritis of the shoulder: a preliminary study. AB - BACKGROUND: While hylan G-F 20 is an approved therapy for the treatment of knee osteoarthritis, there are few reports of its use in shoulder osteoarthritis. HYPOTHESIS: Hylan G-F 20 can reduce pain and improve function in patients with glenohumeral osteoarthritis. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Thirty consecutive patients with idiopathic glenohumeral osteoarthritis who failed to respond to standard conservative measures were enrolled. Patients received 3 weekly intra-articular hylan G-F 20 injections in their affected shoulder and completed a visual analog scale for pain, the UCLA score, and the Simple Shoulder Test at baseline and at 1, 3, and 6 months after the third injection. RESULTS: Of the 30 patients, 3 withdrew during the washout period before treatment; the remaining patients (17 men and 10 women) had an average age of 62 years. The mean baseline visual analog scale score was 54, UCLA score was 15.7, and Simple Shoulder Test score was 5.7 (of 12 possible "yes" responses). At the 6-month follow-up, hylan G-F 20 significantly improved visual analog scale (mean 30, P < .001), UCLA (mean 24, P < .001), and Simple Shoulder Test (7.6 "yes" responses, P < .001) scores. More patients slept comfortably after treatment (56%) versus before treatment (15%). Nine patients had a visual analog scale improvement >40 points after 6 months. No device-related adverse events were observed. CONCLUSION: Hylan G-F 20 may have a beneficial therapeutic effect on some symptomatic patients with glenohumeral osteoarthritis. PMID- 17395959 TI - Influence of modern studded and bladed soccer boots and sidestep cutting on knee loading during match play conditions. AB - BACKGROUND: The influence of modern studded and bladed soccer boots and sidestep cutting on noncontact knee loading during match play conditions is not fully understood. HYPOTHESIS: Modern soccer boot type and sidestep cutting compared with straight-ahead running do not significantly influence knee internal tibia axial and valgus moments, anterior joint forces, and flexion angles. STUDY DESIGN: Controlled laboratory study. METHODS: Fifteen professional male outfield soccer players undertook trials of straight-ahead running and sidestep cutting at 30 degrees and 60 degrees with a controlled approach velocity on a Federation Internationale de Football Association (FIFA) approved soccer surface. Two bladed and 2 studded soccer boots from 2 manufacturers were investigated. Three dimensional inverse dynamics analysis determined externally applied internal/external tibia axial and valgus/varus moments, anterior forces, and flexion angles throughout stance. RESULTS: The soccer boot type imparted no significant difference on knee loading for each maneuver. Internal tibia and valgus moments were significantly greater for sidestep cutting at 30 degrees and 60 degrees compared with straight-ahead running. Sidestep cutting at 60 degrees compared with straight-ahead running significantly increased anterior joint forces. CONCLUSION: Varying soccer boot type had no effect on knee loading for each maneuver, but sidestep cutting significantly increased internal tibia and valgus moments and anterior joint forces. CLINICAL RELEVANCE: Sidestep cutting, irrespective of the modern soccer boot type worn, may be implicated in the high incidence of noncontact soccer anterior cruciate ligament injuries by significantly altering knee loading. PMID- 17395960 TI - Using patient care quality measures to assess educational outcomes. AB - OBJECTIVES: To report the results of a project designed to develop and implement a prototype methodology for identifying candidate patient care quality measures for potential use in assessing the outcomes and effectiveness of graduate medical education in emergency medicine. METHODS: A workgroup composed of experts in emergency medicine residency education and patient care quality measurement was convened. Workgroup members performed a modified Delphi process that included iterative review of potential measures; individual expert rating of the measures on four dimensions, including measures quality of care and educational effectiveness; development of consensus on measures to be retained; external stakeholder rating of measures followed by a final workgroup review; and a post hoc stratification of measures. The workgroup completed a structured exercise to examine the linkage of patient care process and outcome measures to educational effectiveness. RESULTS: The workgroup selected 62 measures for inclusion in its final set, including 43 measures for 21 clinical conditions, eight medication measures, seven measures for procedures, and four measures for department efficiency. Twenty-six measures met the more stringent criteria applied post hoc to further stratify and prioritize measures for development. Nineteen of these measures received high ratings from 75% of the workgroup and external stakeholder raters on importance for care in the ED, measures quality of care, and measures educational effectiveness; the majority of the raters considered these indicators feasible to measure. The workgroup utilized a simple framework for exploring the relationship of residency program educational activities, competencies from the six Accreditation Council for Graduate Medical Education general competency domains, patient care quality measures, and external factors that could intervene to affect care quality. CONCLUSIONS: Numerous patient care quality measures have potential for use in assessing the educational effectiveness and performance of graduate medical education programs in emergency medicine. The measures identified in this report can be used as a starter set for further development, implementation, and study. Implementation of the measures, especially for high stakes use, will require resolution of significant measurement issues. PMID- 17395961 TI - Spirituality, religion, and health. PMID- 17395962 TI - Measuring children's health care quality. PMID- 17395963 TI - The impact of percutaneous coronary interventions on outcomes for acute myocardial infarction in Pennsylvania. AB - This research estimates the benefits associated with percutaneous coronary interventions (PCIs) for patients with acute myocardial infarction (AMI) treated at hospitals in Pennsylvania. We studied 31 351 patients with AMI in Pennsylvania during the year 2000, including 10 170 who received PCI. Univariate comparisons between groups were made using chi2 tests for categorical outcomes and Student's t tests for continuous outcomes. A logit model for proportions was used to model the relationship between mortality and the proportion of AMI patients who received PCI. The mortality rate for patients undergoing PCI was significantly lower than for those being treated medically (1.4% vs 15.8%, P<.0001). Furthermore, significant survival benefits associated with PCI persisted when patients were stratified by age, sex, type of infarction, and severity at admission. At the hospital level, higher rates of PCI were associated with a significantly lower overall mortality rate among patients with AMI (P<.0001). PMID- 17395964 TI - Improvement of psychiatric ambulatory follow-up care by use of care coordinators. AB - This study examined whether patients discharged from inpatient psychiatric care would improve rates of follow-up appointments when designated staff (ie, care coordinators) were assigned to coordinate care after hospital discharge. Data were collected from 1313 psychiatric discharges in 2003 and 1804 psychiatric discharges in 2004, principally from hospitals in the Northeast states. Patients' health care was managed by Anthem Behavioral Health-Northeast. Appointment verification was made through Health Plan Employer Data and Information Set methodology. Of the 1804 psychiatric discharges, 71.6% kept an outpatient appointment within 7 days of discharge, and 88.3% kept an outpatient appointment within 30 days of discharge. These rates were a statistically significant improvement ( P>.001) from the prior year's rates of 66.6% and 84.0%, respectively, when care coordinators were not used. PMID- 17395965 TI - Redefining hospital readmissions to better reflect clinical course of care for heart failure patients. AB - The prevailing definition of a hospital readmission is a hospital admission that occurs within a specified time frame after discharge from the first or index admission. This definition may be questionable for studies with long or indefinite time frames during which the effect of interventions in the index admission will likely wane, making it less appropriate to classify a later hospitalization as a readmission. The current study proposes an alternative definition of readmission and compares the new definition to the traditional one. The comparison shows that the new method is more conceptually correct because it takes into account the waning effect of the index admission. It can identify readmissions regardless of when they occur. It increases a readmission sample size and thus statistical power. PMID- 17395966 TI - A 10-Rights framework for patient care quality and safety. AB - Recent reports in the lay and professional press document the failings of our patient care systems and have led to a multitude of suggestions for patient care quality and safety improvement initiatives. Given the complexity and range of services being offered, hospitals are launching numerous improvement initiatives in nearly all clinical care and support areas. This article describes a quality improvement framework, the "10 Rights," designed to help leaders better understand, organize, and prioritize patient care quality and safety issues and approaches. In addition to describing the framework, each Right is linked to 3 current national efforts at enhancing patient care quality and safety: the Joint Commission on Accreditation of Healthcare Organizations' National Patient Safety Goals, the National Quality Forum 30 Safe Practices, and the Centers for Medicare and Medicaid Services Hospital Quality Measures. PMID- 17395967 TI - Educating to improve patient care: integrating quality improvement into a medical school curriculum. AB - A curriculum focused on disease and the management of disease is no longer adequate for training physicians who are expected to practice in an environment where medical errors, system shortcomings, and physician and practice scorecards are the norm. This article describes a longitudinal curriculum on quality improvement that was integrated into the existing curriculum at Mayo Medical School, from years I through IV, through collaboration with 13 different course and clerkship directors. The curriculum is being taught through exercise-based discussions, video sessions followed by debriefing, simulations, case-based discussions, and lectures. This curriculum prepares physicians in training to be knowledgeable about common systems issues and medical errors in clinical practice and has the potential for application in undergraduate medical education. PMID- 17395968 TI - Diagnostic utility of an epilepsy monitoring unit. AB - Guidelines recommend that patients with persist- ent seizures not responsive to standard antiepileptic drugs after 1 year be referred to specialized epilepsy centers for evaluation, which may include inpatient video-electroencephalographic monitoring. This 1-year retrospective, limited-focus review of 213 admissions and 3-year postdischarge review of epilepsy surgery referrals sought to determine if admission to an inpatient adult epilepsy monitoring unit resulted in a definitive diagnosis, a change in diagnosis or treatment, and whether referral patterns were consistent with guidelines. The median duration of time since symptom onset prior to admission was 15 years. At discharge, 87.8% of admissions received a definitive diagnosis. Epilepsy was diagnosed in 73.3% and excluded in 21.6%. Admissions without epilepsy had been treated with antiepileptic drugs for a median of 9 years. Admission resulted in change in treatment in 79%. This review documents the diagnostic utility of an epilepsy monitoring unit while highlighting that time since symptom onset to admission exceeded established guidelines. PMID- 17395969 TI - Emergency departments in crisis: implications for disaster preparedness. PMID- 17395970 TI - Quality in the delivery of preventive services: The National Colorectal Cancer Roundtable. PMID- 17395971 TI - Early versus delayed endocrine therapy for prostate cancer. AB - Endocrine treatment (ET) has in the past been shown to be beneficial in delaying clinical progression in all stages of prostate cancer, leading to an improvement of progression free survival in virtually all trials ever conducted. The first observations on this issue date back to the studies of the Veterans Administration Cooperative Urological Research Group in the 1960s. The period of time during which ET and the resulting side effects can be avoided is strongly dependent on the clinical stage of the disease. This treatment period is long in men who have minimal disease, such as a rising prostate-specific antigen after potentially curative management; however, it is considerably shorter in men who initially present with metastatic disease. In these situations, the potential benefit in quality of life, and avoidance of adverse events must be matched against the benefit in terms of gaining progression free time for the individual patient. This difficult task is supported by information supplied in this review. Locally advanced and regional (lymph node positive; stage T3N0-1M0Gx) disease is the domain of adjuvant ET. In this field, important progress has recently been made due to trials, which combine aggressive treatment of the primary tumor with adjuvant ET initiated at the same time. Therefore, in locally advanced and regional disease, radiotherapy or surgery combined with adjuvant ET must be considered state-of-the-art. PMID- 17395972 TI - Adrenocortical cancer: pathophysiology and clinical management. AB - Adrenocortical cancer (ACC) is a rare tumor with a poor prognosis. By contrast, benign adrenocortical tumors are frequent, underlying the importance of a correct diagnosis of malignancy of such tumors. ACC can be diagnosed by the investigation of endocrine signs of steroid excess, symptoms due to tumor growth or an adrenal incidentaloma. Hormonal investigations demonstrate in most ACC steroid oversecretion, the dominant characteristics being a co-secretion of cortisol and androgens. Imaging by CT-scan or MRI shows a large heterogeneous tumor with a low fat content. Careful pathological investigation with the assessment of the Weiss score is important for the diagnosis of malignancy. Molecular markers can also be helpful and in the future might be important for prognosis. Tumors localized to the adrenal gland (McFarlane stages 1 and 2) have a better outcome than invasive and metastatic tumors (stages 3 and 4). Tumor removal by a specialized team is crucial for treatment and should always aim at complete removal. In patients with metastatic or progressive disease, medical treatment is started with mitotane that requires a close monitoring of its blood level. Surgery is indicated when possible for local recurrence but also in some cases of metastasis. Local treatment (radiofrequency, chemoembolization, and radiation therapy) can have some indications for metastatic disease. In patients with disease progression cytotoxic chemotherapy can be used. Despite the best care, the overall prognosis of ACC is poor with a 5-year survival rate below 30% in most series. Therefore, progress in the understanding of the pathophysiology of ACC is important. Despite the rarity of ACC, significant advances have been made in the understanding of its pathogenesis the last decade. These progresses came mainly from the study of the genetics of ACC, both at the germline level in rare familial diseases, and at the somatic level by the study of molecular alterations in sporadic tumors. These advances underline the importance of genetic alterations in ACC development and point-out to various chromosomal regions (2, 11p15, 11q, 17p13) and genes (IGF II, p53, beta-catenin, ACTH receptor). This review will summarize these advances as well as the current clinical management of ACC. PMID- 17395973 TI - Staging systems for follicular thyroid carcinoma: application to 171 consecutive patients treated in a tertiary referral centre. AB - A number of risk-group stratification or staging systems have been found useful at stratifying patients with differentiated thyroid carcinoma into risk groups. Those identified as high risk could be subjected to more aggressive treatment, while those at low risk could be spared of such treatment. However, the best stratification system in patients with follicular thyroid carcinoma (FTC) remains unclear. Through a comprehensive MEDLINE search from 1965 to 2005, a total of 18 different staging systems were identified in the literature and 14 of them were applicable to 171 patients, with FTC managed at our institution from 1961 to 2001. Cancer-specific survivals (CSS) were calculated by Kaplan-Meier method and were compared by log-rank test. Using Cox proportional hazards analysis, the relative importance of each staging system in determining CSS was calculated by the proportion of variation in survival time explained (PVE). CSS were predicted by 13 out of the 14 staging systems significantly (P < 0.001). The three highest ranked staging systems by PVE were the new American Joint Committee on Cancer/Union Internationale Centre le Cancer 6th edition, tumour, node, metastases (TNM; 22.4), followed by the Clinical Class (21.2) and the metastases, age, completeness of resection, invasion, size (MACIS; 20.4). In conclusion, 13 out of the 14 presently available staging systems predicted CSS significantly in FTC. When predictability was measured by PVE, the TNM system was found to have the best predictability and thus, should be the stratification system of choice for FTC in the future. PMID- 17395974 TI - p53 family proteins in thyroid cancer. AB - At variance with other human malignancies, p53 mutations are not frequent in thyroid cancer and are believed to be responsible mainly for cancer progression to poorly differentiated and aggressive phenotype. p63 and p73, two proteins with a high degree of homology with p53, are overexpressed in thyroid cancer, but their role in cancer initiation or progression is controversial. Regulation of p53 family protein function depends on: (1) the balance between the expression of transcriptionally active (p53, TAp63, and TAp73) and inactive isoforms (DeltaNp63 and DeltaNp73); (2) their interaction and competition at DNA-responsive elements; (3) their interaction with regulatory proteins, either inhibitory or activating. In thyroid cancer, therefore, although mutations of the p53 oncosuppressor protein family are rare, other mechanisms are present, including aberrant expression of p53 family dominant negative isoforms, up-regulation of inhibitory proteins, and functional inhibition of activating proteins. The overall result is a defective oncosuppressor activity. These inactivating mechanisms may be present in the early stages of thyroid cancer and in different cancer histotypes. A better understanding of this complex network may not only ameliorate our comprehension of cancer biology, but also open the possibility of innovative diagnostic procedures and the development of targeted therapies. PMID- 17395975 TI - Osteoblast-conditioned medium promotes proliferation and sensitizes breast cancer cells to imatinib treatment. AB - Inhibition of platelet-derived growth factor receptor (PDGFR) signaling restricts the growth of human breast cancer in the bone of nude mice. We hypothesized that osteoblast-secreted substances may alter the response capacity of breast cancer cells to the PDGFRs tyrosine kinase inhibitor imatinib mesylate. We found that osteoblast-conditioned medium (OCM) increases the proliferation rate of the estrogen receptor negative (ER-) MDA-MB-231 and of the ER+ MCF-7 human breast cancer cell lines and the growth-promoting effect on ER+ cells is independent from estrogen. OCM significantly improved the dose- and the time-dependent sensitivity of the tumor cells to the anti-proliferative effect of imatinib. We also found that MDA-MB-231 and MCF-7 cells express the two PDGFRs subtypes, PDGFR alpha and PDGFR-beta, and OCM treatment increases the expression of the PDGFRs. Furthermore, imatinib inhibited the phosphorylation rate of its target tyrosine kinase receptors. We conclude that bone microenvironment, through osteoblast secreted substances may cause estrogen-independent proliferation of breast cancer cells by a mechanism mediated by the induction of PDGFRs expression. The enhanced sensitivity of OCM-treated breast cancer cells to imatinib would justify investigation on the efficacy of imatinib in bone breast cancer metastasis. PMID- 17395976 TI - The ubiquitin-binding protein p62 is expressed in breast cancers showing features of aggressive disease. AB - p62 is a multi-functional protein, which induces nuclear factor-kappaB (NFkappaB) activation through multiple upstream signalling pathways, including those triggered by the epidermal growth factor (EGF) family of receptors. We hypothesised that p62 overexpression increased EGF family receptor expression and worse outcome in breast cancer would be associated. We stained a tissue microarray representing 523 breast cancers using a commercial guinea pig anti human p62 sera and standard immunohistochemical methods to address this. Out of n = 106 tumours, 20.3% stained positively. p62 expression correlated with grade (P = 0.010) and distant metastasis (P = 0.04) and EGF receptor (EGFR) (P = 0.012), HER2 (P = 0.016), HER3 (P = 0.007) and HER4 (0.002) expressions. Though expression correlated with reduced 5-year survival (58.5 vs 73.6%), there was no association with overall disease specific survival. p62 expression may represent a marker of activation of the NFkappaB pathway. PMID- 17395978 TI - Pasireotide, a multiple somatostatin receptor subtypes ligand, reduces cell viability in non-functioning pituitary adenomas by inhibiting vascular endothelial growth factor secretion. AB - Somatostatin (SRIF) analogs have been employed in medical therapy of non functioning pituitary adenomas (NFA), with contrasting results. Previous evidence showed that SRIF can exert its antiproliferative effects by reducing vascular endothelial growth factor (VEGF) secretion and action, and that VEGF expression may be related to pituitary tumor growth. The aim of our study was to clarify the possible effects of a multireceptor SRIF ligand on VEGF secretion and cell proliferation in human NFA primary cultures. We assessed the expression of SRIF receptors (SSTR1-5), the in vitro effects on VEGF secretion, and on cell viability of SRIF and of the stable SRIF analog pasireotide (SOM230), which activates SSTR1, 2, 3, and 5. Twenty-five NFA were examined by RT-PCR for expression of alpha-subunit, SSTR, VEGF, and VEGF receptors 1 (VEGF-R1) and 2 (VEGF-R2). Primary cultures were tested with SRIF and with pasireotide. All NFA samples expressed alpha-sub, VEGF and VEGFR-1 and 2, while SSTR expression pattern was highly variable. Two different groups were identified according to VEGF secretion inhibition by SRIF. VEGF secretion and cell viability were reduced by SRIF and pasireotide in the 'responder' group, but not in the 'non-responder' group, including NFA expressing SSTR5. SRIF and pasireotide completely blocked forskolin-induced VEGF secretion. In addition, SRIF and pasireotide completely abrogated the promoting effects of VEGF on NFA cell viability. Our data demonstrate that pasireotide can inhibit NFA cell viability by inhibiting VEGF secretion, and suggest that the multireceptor-SSTR agonist pasireotide might be useful in medical therapy of selected NFA. PMID- 17395977 TI - Serum IGF-I, its major binding protein (IGFBP-3) and epithelial ovarian cancer risk: the European Prospective Investigation into Cancer and Nutrition (EPIC). AB - We set out to study the relationship between circulating levels of IGF-I and its major binding protein (IGFBP-3) in relation to ovarian cancer risk. We conducted a case-control study nested within the European Prospective Investigation into Cancer and Nutrition. Levels of IGF-I and IGFBP-3 were measured in prediagnostic serum samples of 214 women who subsequently developed ovarian cancer, and 388 matched control subjects. Conditional logistic regression models were used to estimate relative risks of ovarian cancer by tertiles of IGF-I and IGFBP-3 levels. For all women, there was no association between the circulating IGF-I or IGFBP-3 levels and the risk of ovarian cancer. However, among women diagnosed with ovarian cancer aged 55 or younger, the relative risk was higher in the middle or top tertiles of serum IGF-I, when compared with women in the lowest tertile (odds ratios (OR) = 1.8 (95%CI 0.7-4.3) and OR = 2.4 (95%CI 0.9-6.4); P(trend) = 0.08) respectively. These results were adjusted for body mass index, previous hormone use, fertility problems, and parity. Restricting the analysis to women who were premenopausal at blood donation, relative risks for ovarian cancer diagnosed before age 55 were higher (OR = 5.1 (95%CI 1.5-18.2) and OR = 5.6 (95%CI 1.5-20.8) respectively, for second and third tertiles; P(trend) = 0.02). Adjustment for serum IGFBP-3 levels only slightly attenuated relative risk estimates. Relations between IGFBP-3 and ovarian cancer before age 55 were in the same direction as for IGF-I, but less strong and statistically not significant. In women aged over 55, there was no association between serum IGF-I or IGFBP-3 and ovarian cancer risk. Our results suggest that the circulating levels of IGF-I may play a potentially important role in the development of ovarian cancer in women of a pre- or perimenopausal age. PMID- 17395979 TI - Incidence and familial risks in pituitary adenoma and associated tumors. AB - Reliable data on familial risks are important for clinical counseling and cancer genetics. We wanted to study incidence trends and familial risks for pituitary adenomas and associated tumors through parental and sibling probands, using the nation-wide Swedish Family-Cancer Database on 10.5 million individuals, containing families with parents and offspring. Cancer data were retrieved from the Swedish Cancer Registry from years 1958 to 2002, including 3239 pituitary tumor patients. Familial risk for offspring was defined through standardized incidence ratio (SIR), adjusted for many variables. The incidence of pituitary adenoma has increased moderately from 1958 to the 1990s and declined thereafter. There were only three offspring-parent pairs with a concordant pituitary tumor, the SIR was not significant. Parental skin cancer (SIR 1.60) and leukemia (1.90, chronic lymphatic leukemia 2.59) were associated with offspring pituitary adenoma diagnosed at any age up to 70 years. There was a strong association of pituitary adenomas with nervous system hemangiopericytomas, SIR 182. The only significant association among siblings was between pituitary tumors and breast cancer (1.46). The risk of pituitary adenoma was marginally increased in individuals whose siblings were diagnosed with colorectal cancer. The results suggest an association of pituitary adenomas with nervous system hemangiopericytomas and breast and colorectal cancers, in addition to some other tumor types. Whether these associations can be explained by the recently identified pituitary adenoma predisposing gene, AIP, remains to be established. PMID- 17395980 TI - Src family kinase activity regulates adhesion, spreading and migration of pancreatic endocrine tumour cells. AB - Pancreatic endocrine tumours (PETs) are rare and 'indolent' neoplasms that usually develop metastatic lesions and exhibit poor response to standard medical treatments. Few studies have investigated pathways responsible for PET cell growth and invasion and no alternative therapeutic strategies have been proposed. In a recent microarray analysis for genes up-regulated in PETs, we have described the up-regulation of soluble Src family tyrosine kinases in this neoplasia, which may represent potentially promising candidates for therapy. Herein, we have investigated the expression and function of Src family kinases in PETS and PET cell lines. Western blot analysis indicated that Src is highly abundant in the PET cell lines CM and QGP-1. Immunohistochemistry and Western blot analyses showed that Src is up-regulated also in human PET lesions. Pharmacological inhibition of Src family kinases by the specific inhibitor PP2 strongly interfered with adhesion, spreading and migration of PET cell lines. Accordingly, the actin cytoskeleton was profoundly altered after inhibition of Src kinases, whereas even prolonged incubation with PP2 exerted no effect on cell cycle progression and/or apoptosis of PET cells. A transient increase in tyrosine phosphorylation of a subset of proteins was observed in QGP-1 cells adhering to the plate, with a peak at 75 min after seeding, when approximately 80% of cells were attached. Inhibition of Src kinases caused a dramatic reduction in the phosphorylation of proteins with different molecular weight that were isolated from the cell extracts by anti-phosphotyrosine immunoprecipitation or pull-down with the SH2 domain of Src. Among them, the docking protein p130Cas interacted with Src and is a major substrate of the Src kinases in QGP-1 cells undergoing adhesion. Our results suggest that Src kinases play a specific role during adhesion, spreading and migration of PET cells and may indicate therapeutical approaches directed to limiting the metastatic potential of these cells. PMID- 17395981 TI - The Ras effectors NORE1A and RASSF1A are frequently inactivated in pheochromocytoma and abdominal paraganglioma. AB - NORE1A (RASSF5) and RASSF1A are newly described Ras effectors with tumour suppressor functions. Both molecules are frequently inactivated in various cancers. In this study, we aimed to explore the potential involvement of NORE1A and RASSF1A in pheochromocytoma and abdominal paraganglioma tumorigenesis. A panel of 54 primary tumours was analysed for NORE1A and RASSF1A mRNA expression by TaqMan quantitative RT-PCR. Furthermore, NORE1A and RASSF1A promoter methylation was assessed by combined bisulphite restriction endonuclease assay and methylation-sensitive Pyrosequencing respectively. The anti-tumorigenic role of NORE1A was functionally investigated in Nore1A-transfected PC12 rat pheochromocytoma cells by fluorescent inhibition of caspase activity and soft agar assays. Significantly suppressed NORE1A and RASSF1A mRNA levels were detected in primary tumours compared with normal adrenal medulla (P<0.001). Methylation of the NORE1A promoter was not observed in primary tumours. On the other hand, 9% (5/54) of the primary tumours examined showed RASSF1A promoter methylation greater than 20% as detected by Pyrosequencing. Methylation of the RASSF1A promoter was significantly associated with malignant behaviour (P<0.05). Transient expression of Nore1a resulted in enhanced apoptosis and impaired colony formation in soft agar. Our study provides evidence that NORE1A and RASSF1A are frequently suppressed in pheochromocytoma and abdominal paraganglioma. Silencing of NORE1A contributes to the transformed phenotype in these tumours. PMID- 17395982 TI - The association between primary hyperparathyroidism and malignancy: nationwide cohort analysis on cancer incidence after parathyroidectomy. AB - In order to evaluate the link between primary hyperparathyroidism (pHPT) and malignancies, cases subjected to parathyroid adenomectomy (PTX) during 1958-1997 in Sweden were identified by analyzing the National Swedish Cancer Registry. To minimize the influence of confounding by detection, cases with malignant disease diagnosed before or at the same time as pHPT or during the first year after PTX were excluded. Altogether 9782 cases (7642 female) were included and followed for up to 40 years. Thus, the study comprises 89,571 person-years of observation. The incidence of malignancies was compared with that in the Swedish population standardized for age, sex, and calendar year. An increased overall incidence of cancer was demonstrated in both genders (standardized incidence ratio (SIR) 1.43, 95% confidence interval (CI) 1.35-1.52). This remain unchanged beyond 15 years after PTX. Breast cancer contributed a quarter of the cancer incidence in women (SIR 1.44, 95% CI 1.25-1.62). An increased risk of kidney (SIR 2.40, 95% CI 1.72 3.25), colonic (SIR 1.46, 95% CI 1.19-1.77), and squamous cell skin cancer (SIR 2.79, 95% CI 2.25-3.43) was found in both genders. The risk of endocrine and pancreas cancer was increased in the minority of patients who had their PTX before the age of 40. We conclude that pHPT is associated with an increased risk of developing malignancies that persists even after PTX. This suggests a causal disassociation with the biochemical derangements caused by parathyroid adenoma, while potentially common etiological mechanisms may include genetic predisposition or acquired disability to withstand environmental influence. PMID- 17395983 TI - The inhibition of Wnt/beta-catenin signalling by 1alpha,25-dihydroxyvitamin D3 is abrogated by Snail1 in human colon cancer cells. AB - The Wnt/beta-catenin signalling pathway is activated in 90% of human colon cancers by nuclear accumulation of beta-catenin protein due to its own mutation or to that of adenomatous polyposis coli. In the nucleus, beta-catenin regulates gene expression promoting cell proliferation, migration and invasiveness. 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) inhibits beta-catenin signalling by inducing its binding to vitamin D receptor (VDR) and by promoting beta-catenin nuclear export. The transcription factor Snail1 represses VDR expression and we demonstrate here that Snail1 also abolishes the nuclear export of beta-catenin induced by 1,25(OH)(2)D(3) in SW480-ADH cells. Accordingly, Snail1 relieves the inhibition exerted by 1,25(OH)(2)D(3) on genes whose expression is driven by beta-catenin, such as c-MYC, ectodermal-neural cortex-1 (ENC-1) or ephrin receptor B2 (EPHB2). In addition, Snail1 abrogates the inhibitory effect of 1,25(OH)(2)D(3) on cell proliferation and migration. In xenografted mice, Snail1 impedes the nuclear export of beta-catenin and the inhibition of ENC-1 expression induced by EB1089, a 1,25(OH)(2)D(3) analogue. The elevation of endogenous SNAIL1 protein levels reproduces the effect of an ectopic Snail1 gene. Remarkably, the expression of exogenous VDR in cells with high levels of Snail1 normalizes the transcriptional responses to 1,25(OH)(2)D(3). However, this exogenous VDR failed to fully restore the blockage of the Wnt/beta catenin pathway by 1,25(OH)(2)D(3). This suggests that the effects of Snail1 on this pathway are not merely due to the repression of VDR gene. We conclude that Snail1 is a positive regulator of the Wnt/beta-catenin signalling pathway in part through the abrogation of the inhibitory action of 1,25(OH)(2)D(3). PMID- 17395984 TI - Role of ERbeta palmitoylation in the inhibition of human colon cancer cell proliferation. AB - The cellular functions regulated by 17beta-estradiol (E2) start after the hormone binds to its receptors (i.e., ERalpha and ERbeta). These act as ligand-dependent transcription factor transactivating target genes. In addition, E2 induces non genomic actions, whose activation is triggered by a fraction of the ERs localized at the plasma membrane. Palmitoylation allows ERalpha to localize at the plasma membrane, to associate with caveolin-1, and, upon E2 stimulation, to activate rapid signals relevant for cell proliferation. The existence of a mechanism, which allows ERbeta localization at the plasma membrane and its putative role in anti-proliferative E2 effects is completely unknown. Here, the susceptibility of ERbeta to undergo palmitoylation and the role played by this process has been analyzed in DLD-1 containing endogenous ERbeta or in HeLa cells transiently transfected with ERbeta or ERalpha expression vectors. As for ERalpha, palmitoylation is necessary for ERbeta localization at the plasma membrane and its association with caveolin-1 but, in contrast to ERalpha, the E2 binding increases ERbeta association with caveolin-1 and the p38 member of MAPK family. Moreover, the palmitoyl acyl transferase (PAT) inhibitor blocks the ability of ERbeta-E2 complex to activate p38 impairing the receptor-dependent activation of downstream proapoptotic cascade (i.e., caspase-3 activation and poly(ADP ribose)polymerase (PARP) cleavage). Consequently, palmitoylation must be considered to be a molecular device for ERbeta, which allows these receptors to interact with the plasma membrane and to regulate E2-induced non-genomic functions relevant to the anti-proliferative effect of this hormone. PMID- 17395985 TI - Hypochlorous acid-mediated generation of glycerophosphocholine from unsaturated plasmalogen glycerophosphocholine lipids. AB - The myeloperoxidase-derived metabolite hypochlorous acid (HOCl) promotes the selective cleavage of plasmalogens into chloro fatty aldehydes and 1 lysophosphatidylcholine (LPC). The subsequent conversion of the initially generated LPC was investigated in plasmalogen samples in dependence on the fatty acid residue in the sn-2 position by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry and (31)P NMR spectroscopy. Plasmalogens containing an oleic acid residue in the sn-2 position are converted by moderate amounts of HOCl primarily to 1-lyso-2-oleoyl-sn-glycero-3 phosphocholine and at increased HOCl concentrations to the corresponding chlorohydrin species. In contrast, plasmalogens containing highly unsaturated docosahexaenoic acid yield upon HOCl treatment 1-lyso-2-docosahexaenoyl glycerophosphocholine and glycerophosphocholine. The formation of the latter product denotes a novel pathway for the action of HOCl on plasmalogens. PMID- 17395986 TI - Skin tears: a review of the evidence to support prevention and treatment. AB - Physiological changes that occur as part of the aging process put the elderly at risk for developing skin tears. Lack of consistent care in the management of skin tears in the authors' community sparked a literature search to determine best evidence for the management of skin tears. Medline and Cochrane Library databases were searched for studies and systematic reviews on skin tear prevention and treatment. The literature suggested that the greatest number of skin tears occurs among the elderly (65 years and older). Factors reported to be associated with the occurrence of skin tears in addition to age include immobility and a history of skin tears. In the absence of systematic risk factor research, clinicians continue to report that all variables that may be associated with these wounds must be considered when assessing the elderly. Several small studies suggest that most skin tears occur on the extremities and seasonal incidence variations also have been reported. Despite consistent sample-size limitations, the results of controlled clinical studies and case series consistently suggest that education and implementation of prevention protocols reduce the incidence of skin tears in extended care facilities by almost 50%. Studies to evaluate and compare the effectiveness and cost-effectiveness of skin tear treatments are limited but when appropriate care is provided most skin tears can be expected to heal after 7 to 21 days depending on the severity of the wound. A combination of skin protection, preventive measures, and treatments that facilitate moist wound healing appear to be most effective. Risk factor and epidemiological studies as well as research to validate the existing skin tear classification system and improve clinician ability to provide evidence-based risk assessments, preventive care, and treatment are needed. PMID- 17395987 TI - The effect of noncontact, low-intensity, low-frequency therapeutic ultrasound on lower-extremity chronic wound pain: a retrospective chart review. AB - Pain associated with chronic wounds and related wound care modalities presents a persistent clinical challenge in patient care, yet evidence supporting the effects of interventions on wound pain remains sparse. In response to initial clinical observations that several patients with painful chronic lower-extremity wounds reported a reduction in wound pain shortly after ultrasound therapy was initiated, a retrospective chart review and analysis of reported pain scores was conducted. The records of 15 consecutive patients (eight women, seven men, age range 28 to 88 years) with painful, nonhealing, lower-extremity wounds treated for 2 to 4 weeks with noncontact, low-intensity, low-frequency therapeutic ultrasound were reviewed and recorded pain scores abstracted. Mean pain scores decreased from 8.07 (+/- 1.91) pre-treatment to 1.67 (+/- 1.76) post-treatment (P = 0.0003). No patients reported worsening pain after treatment commenced. This preliminary evidence suggests that prospective, controlled clinical studies to evaluate the effect of this treatment on wound-related pain are warranted. PMID- 17395988 TI - Evaluating the use of hydrogel sheet dressings in comprehensive burn wound care. AB - Comprehensive burn wound management comprises a challenging spectrum of acute, chronic, traumatic, and surgical wounds with a wide range of anatomical locations and depth. When processing of porcine and cadaver skin - items central to burn care management strategies - became problematic at one Hong Kong hospital, a commercially available cost-effective substitute dressing was urgently needed. After reviewing the characteristics and availability of several dressings, hydrogel sheet dressings were evaluated in a range of burn wound applications. Fifty wounds, including skin graft donor sites, acute partial-thickness burns, and excised full-thickness wounds in 30 consecutive patients were managed with the dressing. It also was used as a temporary dressing over meshed autografts and cultured cell applications. When hydrogel is applied, it is nonadherent; nursing staff reported general ease of use. When applying hydrogel to awkward areas (eg, posterior thigh donor sites), assistance was required to stabilize the large sheets while securing retention dressings. Patients reported no pain during and between dressing changes. No adverse events occurred. Clinical outcomes met or exceeded expectations and guidelines for dressing application and removal were developed. Based on the results obtained, prospective, randomized, controlled clinical studies to ascertain the efficacy and effectiveness of this dressing were initiated. Broader exploration of the advantages of hydrogel use in burn wound care is warranted. PMID- 17395989 TI - Antioxidant measurements. AB - Chemical reactions, including oxidation and reduction of molecules, occur in every cell. These reactions can lead to the production of free radicals. Free radicals react with organic substrates such as lipids, proteins, and DNA. Through oxidation free radicals cause damage to these molecules, disturbing their normal function, and may therefore contribute to a variety of diseases. The anti oxidation system, which consists of enzymatic antioxidants and non-enzymatic antioxidants, defends against oxidative stress. The aim of this review is to summarize general aspects of methods to measure the antioxidant defence system all in one (total antioxidant capacity) and discuss a number of methods which are currently used for detection of antioxidant properties. PMID- 17395990 TI - Techniques for early detection of Alzheimer's disease using spontaneous EEG recordings. AB - Alzheimer's disease (AD) is a degenerative disease which causes serious cognitive decline. Studies suggest that effective treatments for AD may be aided by the detection of the disease in its early stages, prior to extensive neuronal degeneration. In this paper, we propose a set of novel techniques which could help to perform this task, and present the results of experiments conducted to evaluate these approaches. The challenge is to discriminate between spontaneous EEG recordings from two groups of subjects: one afflicted with mild cognitive impairment and eventual AD and the other an age-matched control group. The classification results obtained indicate that the proposed methods are promising additions to the existing tools for detection of AD, though further research and experimentation with larger datasets is required to verify their effectiveness. PMID- 17395991 TI - Noninvasive liver-iron measurements with a room-temperature susceptometer. AB - Magnetic susceptibility measurements on the liver can quantify iron overload accurately and noninvasively. However, established susceptometer designs, using superconducting quantum interference devices (SQUIDs) that work in liquid helium, have been too expensive for widespread use. This paper presents a less expensive liver susceptometer that works at room temperature. This system uses oscillating magnetic fields, which are produced and detected by copper coils. The coil design cancels the signal from the applied field, eliminating noise from fluctuations of the source-coil current and sensor gain. The coil unit moves toward and away from the patient at 1 Hz, cancelling drifts due to thermal expansion of the coils. Measurements on a water phantom indicated instrumental errors less than 30 microg of iron per gram of wet liver tissue, which is small compared with other errors due to the response of the patient's body. Liver-iron measurements on eight thalassemia patients yielded a correlation coefficient r = 0.98 between the room temperature susceptometer and an existing SQUID. These results indicate that the fundamental accuracy limits of the room-temperature susceptometer are similar to those of the SQUID. PMID- 17395992 TI - Calculation of the induced voltage at the terminals of cardiac pacemakers submitted to conducted disturbances. AB - This paper presents a method of numerical simulation based on the admittance method which allows us to calculate the induced tension at the terminals of a cardiac pacemaker subjected to conducted disturbances. The physical model used for simulation is an experimental test bed that makes it possible to study in vitro the behavior of pacemakers subjected to electromagnetic disturbances. The studied frequencies correspond to domestic and industrial applications of electricity (50 Hz-500 kHz). The experimental tests are carried out with two different single-chamber pacemakers implanted in a tissue-equivalent phantom and correlated to tests performed in air (without the phantom). Results obtained by numerical simulation are in good agreement with experimental values and that allows us to validate the computer model. Numerical results are used to determine the transfer function between the signal source and the signal induced between the distal lead tip and the case of the implanted device. PMID- 17395993 TI - A robust fetal ECG detection method for abdominal recordings. AB - In this paper, we propose a new method for FECG detection in abdominal recordings. The method consists of a sequential analysis approach, in which the a priori information about the interference signals is used for the detection of the FECG. Our method is evaluated on a set of 20 abdominal recordings from pregnant women with different gestational ages. Its performance in terms of fetal heart rate (FHR) detection success is compared with that of independent component analysis (ICA). The results show that our sequential estimation method outperforms ICA with a FHR detection rate of 85% versus 60% of ICA. The superior performance of our method is especially evident in recordings with a low signal to-noise ratio (SNR). This indicates that our method is more robust than ICA for FECG detection. PMID- 17395994 TI - Effects of high concentration oxygen administration on n-back task performance and physiological signals. AB - This study investigated the effect of 40% oxygen administration on n-back task performance, blood oxygen saturation and heart rate. Five male (25.8 +/- 1.3 years) and five female (23.0 +/- 1.0 years) college students were selected as the subjects for this study. The experiment consisted of two runs: one was an n-back task with normal air (21% oxygen) administered and the other was with hyperoxic air (40% oxygen) administered. The experimental sequence in each run consisted of Rest1 (1 min), 0-back task (1 min), 2-back task (2 min) and Rest2 (4 min). Blood oxygen saturation and heart rate were measured throughout the four phases. The results of the n-back behavioural analysis reveal that accuracy rates were enhanced with 40% oxygen administration compared to 21% oxygen. When 40% oxygen was supplied, blood oxygen saturation was increased and heart rate was decreased compared to that with 21% oxygen administration. It is suggested that 40% oxygen can stimulate brain activation by increasing actual blood oxygen concentration in the process of cognitive performance, and hyperoxia makes heart rate decrease. This result supports the hypothesis that 40% oxygen administration would lead to increases in n-back task performance. PMID- 17395995 TI - Real time ECG artifact removal for myoelectric prosthesis control. AB - The electrocardiogram (ECG) artifact is a major noise source contaminating the electromyogram (EMG) of torso muscles. This study investigates removal of ECG artifacts in real time for myoelectric prosthesis control, a clinical application that demands speed and efficiency. Three methods with simple and fast implementation were investigated. Removal of ECG artifacts by digital high-pass filtering was implemented. The effects of the cutoff frequency and filter order of high-pass filtering on the resulting EMG signal were quantified. An alternative adaptive spike-clipping approach was also developed to dynamically detect and suppress the ECG artifacts in the signal. Finally, the two methods were combined. Experimental surface EMG recordings with different ECG/EMG ratios were used as testing signals to evaluate the proposed methods. As a key parameter for clinical myoelectric prosthesis control, the average rectified amplitude of the signal was used as the performance indicator to quantitatively analyze the EMG content distortion and the ECG artifact suppression imposed by the two methods. Aiming at clinical application, the optimal parameter assignment for each method was determined on the basis of the performance using the suite of testing signals with various ECG/EMG ratios. PMID- 17395996 TI - Evaluation of a wearable orthosis and an associated algorithm for tremor suppression. AB - We describe a wearable orthosis and an associated algorithm for the simultaneous assessment and treatment of essential tremor, one of the most common movement disorders in humans involving an overactivity of the olivo-cerebellar pathways. A motor providing effective viscosity is fixed on a wearable orthosis in the upper limbs. The motor is controlled by a personal computer with software processing in real time the position and rate of rotation of the joint detected by a chip gyroscope. The orthosis can be used in a monitoring mode and in an active mode. The range of tremor suppression of the signals above the orthosis operational limit ranges from about 3% (percentile 5) to about 79% (percentile 95) in relation to energy in the monitoring mode. Considering both postural and kinetic, the mean tremor energy decreased from 55.49 +/- 22.93 rad2 s(-3) in the monitoring mode to 15.66 +/- 7.29 rad2 s(-3) in the active mode. Medians of power reduction were below 60% for the wrist and the elbow. In addition to supplying new information on the interactions between kinematics, dynamics and tremor genesis, this non-invasive technique is an alternative to current therapies. This new approach will provide new insights into the understanding of motor control. PMID- 17395997 TI - Nonlinear analysis of heart rate dynamics in hyperthyroidism. AB - Studies on the physiology of the cardiovascular system suggested that the generation of the heart rate signal is governed by nonlinear chaotic dynamics. No study investigated the nonlinear dynamics of heart rate in hyperthyroidism. We examined whether the heart rate dynamics of hyperthyroid patients is different from normal controls by the nonlinear analysis of heart rate variability (HRV) with correlation dimension (CD). Thirty-three hyperthyroid Graves' disease patients (30 females and 3 males; age 31 +/- 1 years, means +/- SE) and 33 sex-, age-, and body mass index-matched normal controls were recruited to receive one channel electrocardiogram recording for 30 min. The CD, an index of complexity, was computed from the sequence of normal R-R intervals by the Grassberger and Procaccia algorithm. Compared to the normal controls, the hyperthyroid patients showed significant reductions (P < 0.001) in the mean R-R interval (hyperthyroid 616 +/- 15 versus control 868 +/- 16 ms), the standard deviation of R-R intervals (25 +/- 2 versus 54 +/- 4 ms) and CD (5.02 +/- 0.11 versus 6.42 +/- 0.16). Our study demonstrated for the first time that hyperthyroid patients and normal controls could be distinguished by CD analysis of HRV. In addition, the decreased CD in hyperthyroid patients implies reduced complexity and impaired tolerance to cardiovascular stresses in hyperthyroidism. This finding helps to explain exercise intolerance and irritability manifested by the hyperthyroid patients. PMID- 17395998 TI - Automatic detection of left ventricular ejection time from a finger photoplethysmographic pulse oximetry waveform: comparison with Doppler aortic measurement. AB - Left ventricular ejection time (LVET) is a useful measure of ventricular performance and preload. The present study explores a novel method of continuous LVET monitoring using a noninvasive finger photoplethysmographic pulse oximetry waveform (PPG-POW). A method for the automatic beat-to-beat detection of LVET from the finger PPG-POW is presented based on a combination of derivative analysis, waveform averaging and rule-based logic. The performance of the detection method was evaluated on 13 healthy subjects during graded head-up tilt. Overall, the correlation between the PPG-POW derived LVET and the aortic flow derived LVET was high and significant (r = 0.897, p < 0.05). The bias was -14 +/- 14 ms (mean +/- SD), and the percentage error was 9.7%. Although these results would not be sufficient to satisfy the requirement for clinical evaluation of LVET when absolute accuracy was demanded, the strong correlation between the PPG POW LVET and the aortic LVET on an intra-subject basis (r = 0.945 +/- 0.043, mean +/- SD) would support the application of PPG-POW to detect the directional change in LVET of an individual. This could be very useful for the early identification of progressive hypovolaemia or blood loss. The present study has demonstrated a promising approach to extract potentially useful information from a noninvasive, easy-to-obtain signal that could be readily acquired either from existing patient monitoring equipment or from inexpensive instrumentation. More extensive investigation is necessary to evaluate the applicability of the present approach in clinical care monitoring. PMID- 17396000 TI - Historical perspectives on the clinical development of bisphosphonates in the treatment of bone diseases. AB - Bisphosphonates (formerly termed diphosphonates) were first synthesized in the late 1800s; however, their clinical use has been relatively recent. The bisphosphonates' affinity for hydroxyapatite crystal surface led Procter and Gamble to test these compounds in dental, then medical applications. With key input from university researchers, this led to the medical use of the first bisphosphonate, etidronate disodium in 1968 to treat a young patient with myositis ossificans progressiva. Further clinical research led to widespread medical application for the bisphosphonate class including use as a diagnostic in radionuclide bone imaging agents, treatment of osteoporosis, Paget's disease of bone, hypercalcemia of malignancy and metastatic bone disease. The historical development of bisphosphonates provides an excellent example of how observations and knowledge obtained at the basic science level were applied and successfully tested in the clinic. The end result of these efforts has provided health care professionals with diagnostic and therapeutic tools to improve the lives of patients. PMID- 17396001 TI - Thresholding technique for accurate analysis of density and geometry in QCT, pQCT and microCT images. AB - Computed tomography (CT) is widely used in the assessment of bone parameters in live patients and animals as well as bone samples. Quantitative analysis requires the segmentation of the bone from the surrounding tissue, and most segmentation methods rely on some type of thresholding technique. The aim of this communication is to highlight the influence of threshold selection on various bone parameters and recommend appropriate thresholds. Two types of information are of interest in bone analysis from images: geometric parameters and density parameters. We know from imaging theory that blurring is an inherent byproduct of all imaging methods. Depending on the threshold used for segmentation, the object boundary moves in space due to the sloping edge. It is, thus, critical to select the threshold that creates an object boundary that reflects the actual object size. Similarly, due to blurring, the imaged density shows erroneous values at the object boundaries. Such values must not be included for an accurate representation of the object density. Using a pQCT scanner and a bone phantom with known density and geometry, we show that the thresholds for geometry and density are different. The threshold for accurate geometric segmentation was 49% of the difference of the density between the adjacent tissues. The threshold for accurate density assessment was 95% of the maximum density value of the bone. These specific thresholds are valid only for the scanner tested; however, the principle for selecting the thresholds is valid across scanner platforms and scale of imaging. PMID- 17396002 TI - Animals and spaceflight: from survival to understanding. AB - Animals have been a critical component of the spaceflight program since its inception. The Russians orbited a dog one month after the Sputnik satellite was launched. The dog mission spurred U.S. interest in animal flights. The animal missions proved that individuals aboard a spacecraft not only could survive, but also could carry out tasks during launch, near-weightlessness, and re-entry; humans were launched into space only after the early animal flights demonstrated that spaceflight was safe and survivable. After these humble beginnings when animals preceded humans in space as pioneers, a dynamic research program was begun using animals as human surrogates aboard manned and unmanned space platforms to understand how the unique environment of space alters life. In this review article, the following questions have been addressed: How did animal research in space evolve? What happened to animal development when gravity decreased? How have animal experiments in space contributed to our understanding of musculoskeletal changes and fracture repair during exposure to reduced gravity? PMID- 17396003 TI - Genetic and environmental determinants of osteoporosis. AB - Osteoporosis is a common, complex disease that is influenced by genetic and environmental factors. Although molecular genetic studies have identified several potential regions of linkage, underlying susceptibility gene(s) are largely unknown. Genetic susceptibility to osteoporosis may be both context dependent and developmentally regulated, and epigenetic mechanisms are the likely link between gene and environment. In this paper we will review the status of genetic research into osteoporosis, and present the evidence for gene-environment interaction in its pathogenesis. Finally, the current challenges and future directions of research will be briefly discussed. PMID- 17396004 TI - Skeletal responses to space flight and the bed rest analog: a review. AB - The potential for loss of bone mineral mass due to space flight was recognized by space scientists even before man's first venture into micro-gravity. Early life science studies in both the U.S. and Russian space programs attempted to measure the effects of reduced gravity on skeletal homeostasis, and these measurements have become more sophisticated with time. Bone-related measurements have typically included: bone mineral density measured by X-ray absorptiometry and more recently CT scanning; bonerelated hormones and other biochemical markers of bone turnover; and calcium excretion and balance. These measurements, conducted over the last 4 decades, have shed light on the nature of disuse bone loss and have provided preliminary information regarding bone recovery. Ground-based analog (bed rest) studies have provided information complementary to the space flight data and have allowed the testing of various countermeasures to bone loss. In spite of the wealth of knowledge obtained thus far, many questions remain regarding bone loss, bone recovery, and the factors affecting these skeletal processes. This paper will summarize the skeletal data obtained to date by the U.S. and Russian space programs and in ground-based disuse studies. In addition, related body composition data will be briefly discussed, as will possible countermeasures to space flight-induced bone loss. PMID- 17396005 TI - The importance of mechanical loading in bone biology and medicine. AB - This paper discusses the premise that the skeleton is primarily a mechanical organ, and reviews the reasons that mechanical factors play a major role in bone biology. It begins by considering three basic observations: (1) Galileo's observation that bone proportions become more robust as the species' overall size increases; (2) da Vinci's observation that larger structures are inherently weaker than smaller structures subjected to the same stress; and (3) the general observation that each unit of bone mass provides structural support for about 15 units of soft tissue organ mass. Together, these observations lead to the concept that it can be advantageous to minimize bone mass, consistent with constraints on other factors. This premise is discussed here in relation to the phenomenon of bone remodeling, which is seen to serve two purposes: the adjustment of bone mass and geometry to maintain peak bone strains at their maximum tolerable values, and the continual removal of fatigue damage produced at those strain levels. Finally, it is observed that bone remodeling apparently originated approximately 250 million years ago when the first vertebrates of substantial size became weight bearing on land, suggesting that mechanical forces associated with weight-bearing were instrumental in the evolution of bone remodeling. PMID- 17396006 TI - Skeletal fragility and bone quality. PMID- 17396007 TI - Application of skinned single muscle fibres to determine myofilament function in ageing and disease. AB - The chemically skinned fibre is a suitable preparation to determine whether alterations in myofilament function contribute to muscle dysfunction during ageing and disorders such as chronic obstructive pulmonary disease (COPD). In this preparation the sarcolemma is chemically permeabilized and the myofilament lattice kept intact, functioning under controlled near-physiological conditions. As force generating capacity is an important determinant of muscle function and is related to fibre crosssectional area (FCSA), we compared several methods employed by researchers to determine FCSA. Specific tension, force divided by FCSA, has a co-efficient of variation of 27%, 37%, or 30% when the FCSA was measured from the width and depth assuming an elliptical circumference, the width assuming a circular circumference, and the width while the fibre was suspended in the air, respectively. The last method showed the closest relation with the FCSA in histological sections. The velocity of maximal unloaded shortening (V(0)) varied with fibre type, with fibres expressing the Beta/slow (type I) myosin heavy chain (MyHC) isoform being the slowest and fibres expressing the IIb MyHC isoform the fastest. While muscle weakness experienced after surgery could not be explained by changes in specific tension or FCSA of individual fibres, the preparation revealed significant changes in myofilament function during ageing and COPD. PMID- 17396008 TI - Bone loss and mechanical properties of tibia in spinal cord injured men. AB - AIM: The effects of Spinal Cord Injury (SCI) on bone in paralyzed areas are well documented but there are few data for the importance of the level of injury in the decrease of mechanical strength in paralyzed legs. The aim of the present study was to describe bone loss of the separate compartments of trabecular and cortical bone in spinal cord injured men and to compare possible changes in mechanical properties of tibia with the neurological level of injury. MATERIALS AND METHODS: Fifty men were included in this study: 39 had complete SCI in chronic stage. As chronic stage, we considered paraplegia >1.5 years (yrs). Men were separated as follows: Group A (18 men, high paraplegia: Thoracic (T)4-T7 level, mean age: 33 yrs, duration of paralysis: 5.9 yrs) and group B (21 men, low paraplegia: T8-T12 level, mean age: 39 yrs, duration of paralysis: 5.6 yrs) in comparison with 11 healthy men as a control group (C) of similar age, height, and weight. None of the subjects was given bone acting drugs. The neurological profile of each patient was assessed according to the American Spinal Injury Association (ASIA). All subjects were measured by peripheral quantitative computed tomography (pQCT). Measurements were performed at the tibia with a Stratec XCT 3000 (Stratec Medizintechnik, Pforzheim, Germany) scanner. The distal end of the tibia was used as an anatomical marker. The bone parameters, bone mass density (BMD) trabecular, BMD total, BMD cortical, and cortical thickness have been measured at 4% and 38%, respectively, of the tibia length proximal to this point, and the periosteal and endocortical was measured at 14% of the tibia. We calculated stress strain index (SSI), a bone strength estimator derived from the section modulus, and the volumetric density of the cortical area at 14% (SSIPol2) and 38% (SSIPol3) of the tibia length proximal to the distal end of the tibia. RESULTS: In both groups A and B most bone mass parameters were statistically decreased in comparison with controls. In each group we calculated the median deltaSSI(3-2) (SSIPol3 - SSIPol2). In the paraplegic groups Spearman correlation coefficient between duration of paralysis and deltaSSI(3-2) was in group A: r= 0.178, p=N.S. and group B: r=0.534, p=0.027, respectively. CONCLUSION: Despite the similar paralytic effect on bone in all paraplegic patients in our study and because of the non-significant duration of paralysis between paraplegic groups (p=0.87), the two paraplegic groups act differently in mechanical properties of the tibia. In addition, group A patients in respect to the level of injury, are susceptible to autonomic dysreflexia as a result of the disruption of the autonomic nervous system pathways. These results suggest that neurogenic factors are influencing geometric bone parameters. PMID- 17396009 TI - Trabecular bone modeling and subcapital femoral fracture. AB - Fragility fractures, including neck of femur fractures, result from reductions in the amount, quality and architecture of bone. The aim of this study was to compare the cancellous bone structure, and static indices of bone turnover, in female patients who had sustained fragility fracture at the femoral neck, with age-matched females without fragility fracture. Bone samples were taken from the intertrochanteric region of the proximal femur of female patients undergoing hip arthroplasty surgery for a subcapital fragility fracture of the femoral neck (#NOF) or from age-matched female control individuals at routine autopsy. The histomorphometric data, which were normally distributed, indicated no difference between the mean values for any of the structural parameters in control and fracture samples. In particular, the BV/TV values were not different and did not change significantly with age in these cohorts of individuals aged >65 years. The static indices of bone turnover, eroded surface (ES/BS) and osteoid surface (OS/BS), were positively correlated with age in the >65-year-old control group (p<0.05 and p<0.03, respectively). The median values for these indices were not different between the fracture and control groups. However, both the median and the range of OS/BS values were increased for >65-year-old controls compared with a group of younger females aged <65 years, suggesting an increase in bone formation in older females in the proximal femur after 65 years of age. When the data were further interrogated, a reduction in the percentage osteoid surface to eroded surface quotient (OS/ES) was found for the fracture group compared with the age-matched control group. These data indicate that perturbations in bone formation and/or resorption surface are potentially important in producing bone in the proximal femur with increased propensity to fracture. These data also support the concept that trabecular bone modeling may be a factor influencing bone strength in addition to bone mass. PMID- 17396010 TI - Changes in diaphyseal and epiphyseal bone parameters in thoroughbred horses after withdrawal from training. PMID- 17396011 TI - Preliminary results on the mobility after whole body vibration in immobilized children and adolescents. AB - The present article is a preliminary report on the effect of Whole Body Vibration (WBV) on the mobility in long-term immobilized children and adolescents. WBV was applied to 6 children and adolescents (diagnoses: osteogenesis imperfecta, N=4; cerebral palsy, N=1; dysraphic defect of the lumbar spine, N=1) over a time period of 6 months. WBV was applied by a vibrating platform constructed on a tilt table. The treatment effect was measured by alternations of the tilt-angle of the table and with the "Brief assessment of motor function" (BAMF). All 6 individuals were characterized by an improved mobility, which was documented by an increased tilt-angle or an improved BAMF-score. The authors concluded WBV might be a promising approach to improve mobility in severely motor-impaired children and adolescents. Therefore, the Cologne Standing-and-Walking- Trainer powered by Galileo is a suitable therapeutic device to apply WBV in immobilized children and adolescents. PMID- 17396012 TI - Influence of birth weight on peripheral bone in young Gambian adults: a pQCT study. PMID- 17396013 TI - Influence of birth weight and early diet on peripheral bone in premenopausal Cambridge women: a pQCT study. PMID- 17396014 TI - Physical activity in osteopenia treatment improved the mass of bones directly and indirectly submitted to mechanical impact. AB - The effect of physical activity in the treatment of osteopenia induced by ovariectomy was studied in 34 two-month-old Wistar female rats. Animals were divided into three groups in which two were formed by ovariectomized (OVX) animals and the other one had sham-operated animals. Group 1, active OVX'd rats; group 2, sedentary OVX'd rats and group 3, sham-operated ones (control). After three months of daily physical activity in a motor-driven treadmill all rats were sacrificed. In order to perform a histomorphometric analysis, long bones, vertebrae, and nasal bone were selected at necropsy. Ovariectomized rats which exercised showed an increased trabecular bone volume, cortical thickness in the long bones and vertebrae and also an increased nasal bone thickness. Physical activity also increased the connection of osteocytes. It was concluded that physical activity in osteopenia treatment increases and restores the mass of bones directly and indirectly submitted to physical impact. PMID- 17396015 TI - Expressed and cryptic flagellin genes in the H44 and H55 type strains of Escherichia coli. AB - Bacterial H antigens are specified by flagellin molecules, which constitute the flagellar filament. Escherichia coli 781-55 and E2987-73 are the type strains for H44 and H55 antigens, respectively. Unlike E. coli K-12, they possess two flagellin genes, fliC and fllA, on their chromosomes. However, they are monophasic, expressing exclusively the fllA genes, which specify the type antigens. In this study, the flagellin genes were cloned from these strains and their structure and expression were analyzed. It was found that the fliC genes encode apparently intact flagellin subunits but possess inefficient sigma28 dependent promoters, which may result in these genes being silent. The chromosomal locations of the fllA genes are approximately, but not exactly, identical with that of the phase-2 flagellin gene, fljB, of diphasic Salmonella strains. However, unlike the Salmonella fljB gene, the invertible H segment and the fljA gene responsible for the control of flagellar phase variation are both absent from the fllA loci. The fllA genes are highly homologous to the E. coli fliC gene but distantly related to the Salmonella fljB gene. These results suggest a hypothesis that the fllA genes may have emerged by an intra-species lateral transfer of the fliC gene. This hypothesis is further supported by the observation that the fllA genes are flanked by several IS elements and located within cryptic prophage elements. PMID- 17396016 TI - The accurate replacement of long genome region more than several hundreds kilobases in Bacillus subtilis. AB - Competent cell transformation with DNA obtained by the gentle lysis of protoplasts (LP transformation) was used to replace a large genomic region in this study. Discontinuity was detected in the replacement of the donor region tested, probably due to multiple crossover events involving a single donor genome fragment. To overcome discontinuous replacement, we inverted the genomic region to be replaced in the donor used for LP transformation. The replaced region in the transformant was identified to have a continuous genomic region originating from the donor genome. Furthermore, the genome region to be replaced was inverted in the recipient, and the same region and the flanking 10 kb region of both ends was inverted in the donor genome. LP transformation was conducted with the two inversion mutants and it is possible to restrict homologous recombination to the 10 kb flanking regions. Using this method, the 99 kb yxjG-yxbA region, the 249 kb pbpG-yxbA region and the 602 kb yvfT-yxbA region were suggested to be replaced continuously and accurately. PMID- 17396017 TI - Mre11 mediates gene regulation in yeast spore development. AB - Mre11, together with Rad50 and Xrs2/NBS, plays pivotal roles in homologous recombination, repair of DNA double strand breaks (DSBs), activation of damage induced checkpoint, and telomere maintenance. Here we demonstrate that the absence of Mre11 in yeast causes specific effects on regulation of a class of meiotic genes for spore development. Using DNA microarray assays to analyze yeast mutants defective for meiotic DSB formation, we revealed that the meiotic expression profile in the mre11Delta cells was generally unaffected when compared to the one in the wild-type strain, although the activation of about 90 meiotic genes were severely and specifically impaired in early meiosis. These defects were confirmed by northern and lacZ reporter gene assays. Interestingly, a substantial portion of the severely affected genes includes genes responsible for spore wall biogenesis, the defects of which may account for the fragile spore wall phenotype of the mre11Delta strain. The transcriptional deficiency was not observed in other DSB mutants such as rad50Delta, xrs2Delta, spo11Delta, and spo11Y135F, suggesting the transcriptional defect in mre11Delta is due to neither lack of meiotic DSB formation, nor disintegrity of Mre11-Rad50-Xrs2 complex. In addition, the deficiency of mre11Delta in gene activation was not alleviated by the deletion of RAD24. Therefore, it is unlikely that DNA damage checkpoint activation by mre11Delta caused transcriptional deficiency. We also found that a C-terminus DNA binding domain truncation mutant (mre11DeltaC49), which has meiosis-specific defects, exhibited transcriptional defects as observed in mre11Delta, whereas an N-terminal phosphoesterase mutant (mre11D16A) does not. Taken together, we propose that Mre11 is involved in the regulation of a specific class of genes during spore development through its C-terminus domain. PMID- 17396018 TI - Error-free RAD52 pathway and error-prone REV3 pathway determines spontaneous mutagenesis in Saccharomyces cerevisiae. AB - Using the CAN1 gene in haploid cells or heterozygous diploid cells, we characterized the effects of mutations in the RAD52 and REV3 genes of Saccharomyces cerevisiae in spontaneous mutagenesis. The mutation rate was 5-fold higher in the haploid rad52 strain and 2.5-fold lower in rev3 than in the wild type strain. The rate in the rad52 rev3 strain was as low as in the wild-type strain, indicating the rad52 mutator phenotype to be dependent on REV3. Sequencing indicated that G:C-->T:A and G:C-->C:G transversions increased in the rad52 strain and decreased in the rev3 and rad52 rev3 strains, suggesting a role for REV3 in transversion mutagenesis. In diploid rev3 cells, frequencies of can1Delta::LEU2/can1Delta::LEU2 from CAN1/can1Delta::LEU2 due to recombination were increased over the wild-type level. Overall, in the absence of RAD52, REV3 dependent base-substitutions increased, while in the absence of REV3, RAD52 dependent recombination events increased. We further found that the rad52 mutant had an increased rate of chromosome loss and the rad52 rev3 double mutant had an enhanced chromosome loss mutator phenotype. Taken together, our study indicates that the error-free RAD52 pathway and error-prone REV3 pathway for rescuing replication fork arrest determine spontaneous mutagenesis, recombination, and genome instability. PMID- 17396019 TI - Involvement of mitochondrial-targeted RecA in the repair of mitochondrial DNA in the moss, Physcomitrella patens. AB - Homologous recombination is a universal process that contributes to genetic diversity and genomic integrity. Bacterial-type RecA generally exists in all bacteria and plays a crucial role in homologous recombination. Although RecA homologues also exist in plant mitochondria, there have been few reports about the in vivo functions of these homologues. We identified a recA gene orthologue (named PprecA1) in a cDNA library of the moss, Physcomitrella patens. N-terminal fusion of the putative organellar targeting sequence of PpRecA1 to GFP caused a targeting of PpRecA1 to mitochondria. PprecA1 partially complemented the effects of a DNA damaging agent in an Escherichia coli recA deficient strain. Additionally, the expression of PprecA1 was induced by treating the plants with DNA damaging agents. Disruption of PprecA1 by targeted replacement resulted lower rate of the recovery of the mitochondrial DNA from methyl methan sulfonate damage. This is the first report about the characteristics of a null mutant of bacterial-type recA gene in plant. The data suggest that PprecA1 participates in the repair of mitochondrial DNA in P. patens. PMID- 17396020 TI - Close genetic relationship between cultivated and natural populations of common buckwheat in the Sanjiang area is not due to recent gene flow between them-An analysis using microsatellite markers. AB - Natural populations of wild common buckwheat have been found growing adjacent to cultivated populations of common buckwheat. Gene flow between the cultivated and natural populations would be expected in such cases. To evaluate the amount of gene flow, two sets composed of a cultivated buckwheat population and an adjacent natural population of wild common buckwheat were chosen, one from Yanjing village in the Sanjiang area, which is presumed to be the original birthplace of common buckwheat, and one from Jinhe village, Yanyuan district of Sichuan province in China. The genotypes of 45 individuals from each population were examined at eight microsatellite marker loci to estimate the magnitude of gene flow between the cultivated and wild common buckwheat populations. The Bayesian method with a Markov chain Monte Carlo approach estimated that the magnitude of gene flow between the populations in the Sanjiang area at 0.002-0.008 was not significantly different from that found in Yanyuan district at 0.002-0.008. The gene flow between cultivated populations was higher, usually at 0.002-0.044 (exceptionally high at 0.255 between cultivated populations of Yanjing and Jinhe), than that found between a cultivated population and a natural population (0.002-0.008) or between two natural populations (0.002-0.003). Therefore, the genetic similarity found between the cultivated populations and natural populations observed in the Sanjiang area (Konishi et al., 2005) was not due to recent gene flow between them. This in turn suggests that the close genetic relationship found in the Sanjiang area may be due to the common ancestry of the natural populations and cultivated common buckwheat. PMID- 17396021 TI - Distribution of S-alleles in island populations of flowering cherry, Prunus lannesiana var. speciosa. AB - We surveyed the distribution of S-alleles in natural island populations of Prunus lannesiana var. speciosa sampled from seven sites on the Izu Peninsula and six Izu islands, Japan. The S-genotypes of sampled individuals were determined by Southern analysis of RFLPs generated by restriction enzyme digestion of genomic DNA, using cDNA of the S-RNase gene as a probe. All individuals were heterozygous, as expected under gametophytic self-incompatibility (GSI). Sixty three S-alleles were observed in the species, but 12 private to the Izu Peninsula population seemed to be derived from related species, giving a total of 75. The estimated number of S-alleles in each population ranged from 26 to 62, and was inversely correlated with the respective population's distance from the Izu Peninsula, the closest point in the mainland to the islands. This geographical cline in the estimated numbers of S-alleles suggests that gene flow to and from the distant island populations was less frequent, and that the studied species has migrated from the mainland to the Izu islands. The genetic relationship at the S-locus among populations also gave an "isolation by distance" pattern. The genetic differentiation at the S-locus among the populations was very low (F(ST) = 0.014, p < 0.001). The number of S-alleles in the species did not seem to depend on genetic differences associated with population subdivisions. This might be due to the greater effective migration rates of S-alleles, as expected under balancing selection in GSI. PMID- 17396022 TI - Molecular phylogeny of the Drosophila auraria species complex and allied species of Japan based on nuclear and mitochondrial DNA sequences. AB - We studied the phylogenetic relationships among the six species of the Drosophila auraria species complex and three species of the montium species subgroup endemic to Japan. To do this, we examined the DNA sequences of the nuclear ITS1 and the mitochondrial COII of these species. Intraspecific variation in ITS1 was observed for most of the species studied. However, interspecific variation was small and of the same magnitude as intraspecific variation among D. auraria, D. triauraria, and D. quadraria. In addition, the amount of intraspecific variation between COII sequences from D. auraria and D. triauraria was almost the same as that of the interspecific variation between these species. Phylogenetic trees based on ITS1 or COII data reveal that, with the exception of D. pectinifera, these species can be separated into two taxonomic groups. D. rufa formed a taxonomic lineage with two Japanese endemic species, D. asahinai and D. lacteicornis. Physiological and behavioral differences between D. auraria and D. triauraria have been reported, but our results differed from those previously reported. Such differences may be better explained by hybridization between D. auraria and D. triauraria than by similarity due to recent common ancestry. Both the ITS1- and COII-based phylogenetic trees placed D. pectinifera outside the auraria complex. However, D. pectinifera sequences were more similar to those of the auraria complex than those of D. punjabiensis, another species thought to be closely related to members of the auraria complex. PMID- 17396023 TI - Molecular phylogenetic analysis of the human endogenous retrovirus E (HERV-E) family in human tissues and human cancers. AB - The human genome is estimated to contain up to 50 copies of full-length and truncated members of HERV-E family. They are thought to be involved in human gene transcription. Here we examine the expression pattern and phylogenetic relationships of the HERV-E in diverse human tissues and cancer cells using RT PCR amplification and bioinformatic tools. The env gene was expressed in many human tissues (brain, prostate, testis, kidney, placenta, spleen, thymus and uterus) but not in heart, liver, lung and skeletal muscle, importantly, HERV-E expression was detected in all cancer cell lines examined (RT4, PFSK-1, BT-474, HCT-116, TE-1, UO-31, Jurkat, HepG2, A549, MCF7, OVCAR-3, MIA-PaCa-2, PC3, LOX IMVI, AZ521, 2F7, U-937 and C-33A), suggesting that HERV-E family are expressed corresponding to the transcriptional program of human tissues and human cancer cells. Phylogenetic analysis of HERV-E env family from human tissues, cancer cells and our previous data identify two groups (I and II) through evolutionary divergence. Taken together, HERV-E family expression in human tissues and human cancer cells exhibited close relationships of the env gene sequences across human chromosomes. These active HERV-E elements deserve further investigation as potential pathogenic factors in human diseases such as cancers. PMID- 17396024 TI - Different profiles of IL-10+IFN-gamma-IL-4-CD4+ T cells in the peripheral blood in atopic and non-atopic asthmatics. AB - BACKGROUND: The impaired production of interleukin (IL) 10 from regulatory T cells has been proposed as a causal mechanism of asthma. Although IL-10-producing (IL-10+) T cells are detectable in the peripheral blood, their significance in the pathophysiology of asthma remains uncertain. OBJECTIVES: This study aimed to investigate the profile of circulating IL-10+CD4+ T cells in atopic and non atopic asthma. METHODS: Atopic and non-atopic asthmatics were divided into a mild and severe group. Their peripheral blood mononuclear cells (PBMCs) were stimulated with anti-CD3 and anti-CD28 antibodies and then processed for triple cytokine flow cytometry directed to IL-10, interferon (IFN) gamma and IL-4. RESULTS: IL-10+CD4+ cells were exclusively detected in the IFN-gamma-IL-4- population. In atopic asthma, the frequency of IL-10+IFN-gamma-IL-4-CD4+ cells in the severe group was significantly lower than that in the mild group. The frequency of IL-10+IFN-gamma-IL-4-CD4+ cells in the severe group was not significantly different from that in the mild group of those with non-atopic asthma. The frequency of IL-4+IFN-gamma-IL-10-CD4+ cells (Th2) was significantly higher in the group with mild atopic asthma than in that with mild non-atopic asthma. IFN-gamma+IL-4-IL-10-CD4+ cells (Th1) did not differ between groups, irrespective whether the subjects suffered from atopic or non-atopic asthma. CONCLUSIONS: IL-10+CD4+ cells in PBMCs may be distinct from Th1 or Th2 and likely have the profile of regulatory T cells. The differential association of IL-10+IFN gamma-IL-4-CD4+ cells with clinical severity between atopic and non-atopic asthma implies that its pathophysiological significance may differ among asthma phenotypes. PMID- 17396025 TI - Absence of alveolar tears in rat lungs with significant alveolar instability. AB - BACKGROUND: Lung injury associated with the acute respiratory distress syndrome can be exacerbated by improper mechanical ventilation creating a secondary injury known as ventilator-induced lung injury (VILI). We hypothesized that VILI could be caused in part by alveolar recruitment/derecruitment resulting in gross tearing of the alveolus. OBJECTIVES: The exact mechanism of VILI has yet to be elucidated though multiple hypotheses have been proposed. In this study we tested the hypothesis that gross alveolar tearing plays a key role in the pathogenesis of VILI. METHODS: Anesthetized rats were ventilated and instrumented for hemodynamic and blood gas measurements. Following baseline readings, rats were exposed to 90 min of either normal ventilation (control group: respiratory rate 35 min(-1), positive end-expiratory pressure 3 cm H(2)O, peak inflation pressure 14 cm H(2)O) or injurious ventilation (VILI group: respiratory rate 20 min(-1), positive end-expiratory pressure 0 cm H(2)O, peak inflation pressure 45 cm H(2)O). Parameters studied included hemodynamics, pulmonary variables, in vivo video microscopy of alveolar mechanics (i.e. dynamic alveolar recruitment/derecruitment) and scanning electron microscopy to detect gross tears on the alveolar surface. RESULTS: Injurious ventilation significantly increased alveolar instability after 45 min and alveoli remained unstable until the end of the study (electron microscopy after 90 min revealed that injurious ventilation did not cause gross tears in the alveolar surface). CONCLUSIONS: We demonstrated that alveolar instability induced by injurous ventilation does not cause gross alveolar tears, suggesting that the tissue injury in this animal VILI model is due to a mechanism other than gross rupture of the alveolus. PMID- 17396026 TI - The initial response to inhaled nitric oxide treatment for intensive care unit patients with acute respiratory distress syndrome. AB - BACKGROUND: Inhaled nitric oxide (INO) can improve hypoxemia and reduce pulmonary hypertension, but there is a wide range of response to INO. OBJECTIVES: The aim of this study was to evaluate the effect of different INO concentrations in acute respiratory distress syndrome (ARDS) patients. METHODS: Thirty-two adult ARDS patients who were supported by mechanical ventilator were included. INO was given at a concentration of 1, 5, 10, 20 and 40 ppm, sequentially. Arterial blood gas and mean pulmonary artery pressure (MPAP) were measured 30 min after INO concentrations changed. RESULTS: There was a significant increase in PaO(2)/FiO(2) (p < 0.0001) and a decrease in pulmonary vascular resistance index and MPAP (p < 0.0001) after INO use. INO concentrations required for improving oxygenation were in the range of 1-20 ppm, whereas concentrations required for decreasing MPAP were in the range of 1-40 ppm. PaO(2)/FiO(2) worsened when the INO dose was adjusted above 20 ppm. Sixty-nine percent of ARDS patients were responders; 31% of them were nonresponders. Among responders, 64% of patients responded at 1 ppm, 36% at 5 ppm. CONCLUSIONS: The optimal doses for improving oxygenation and reducing PAP differ. The maximum PaO(2)/FiO(2) was observed at a lower INO concentration than that required for the minimal MPAP. There was no further improvement in PaO(2)/FiO(2) when the INO dose was adjusted above 20 ppm. Higher doses of INO treatment worsened oxygenation. PMID- 17396027 TI - Expiratory flow limitation in patients with pleural effusion. AB - BACKGROUND: Expiratory flow limitation (EFL) is one of the main mechanisms contributing to dyspnea in patients with chronic obstructive pulmonary disease but has not been explored in patients with pleural effusion. OBJECTIVES: It was the aim of this study to determine whether patients with pleural effusion exhibit EFL and to investigate the effect of therapeutic thoracentesis on EFL. PATIENTS AND METHODS: The study was performed on 21 patients with pleural effusion who were subjected to thoracentesis and measurement of pleural pressure (PP). Spirometry and estimation of flow limitation by the negative expiratory pressure technique were performed before and after thoracentesis. RESULTS: Statistically significant differences were observed in all spirometric parameters. No correlation between the increase in lung volumes and flows and any of the aspirated fluid parameters was observed. Before thoracentesis, 14 out of 21 patients were flow limited, compared with 7 patients after thoracentesis (chi(2) = 6.151, p = 0.013). Mean values of flow limitation before and after thoracentesis differed significantly. The decrease in flow limitation did not correlate with the increase in the spirometric parameters, the aspirated fluid volume or PP decrease. CONCLUSIONS: In the majority of patients with pleural effusion, flow limitation improves after thoracentesis. Flow limitation may be a contributing factor to the sensation of dyspnea in these patients. PMID- 17396028 TI - Complete tracheal ring. PMID- 17396030 TI - Subtle attentional deficits in the absence of dementia are associated with an increased risk of post-operative delirium. AB - BACKGROUND: Previously, key studies of the risk profile for post-surgical delirium have focused on general medical and non-elective patients, few have examined elective cohorts. Accurate prediction is imperative for clinical trials and prevention strategies. AIMS/HYPOTHESIS: Our hypothesis was that subtle pre operative impairments of attention will be associated with risk of post-operative delirium. METHOD: A prospective study evaluating pre- and post-operative neuropsychological performance in older (> or =70) consecutive elective admissions for orthopaedic surgery, and free of dementia (n = 100) was initiated in a general medical hospital. RESULTS: Pre-operative attentional deficits were closely associated with delirium. Patients who developed post-surgical delirium had significantly slower mean reaction times (p < or = 0.011) and greater variability of reaction time (p = 0.017). A 4- to 5-fold increased risk of delirium was observed for people one standard deviation above the sample means on these variables. CONCLUSIONS: The present study describes a measurement of attentional performance which could form the basis of a neuropsychological marker of delirium. PMID- 17396029 TI - Podocyte injury and glomerulosclerosis in hyperhomocysteinemic rats. AB - BACKGROUND/AIMS: We previously reported that increase in plasma homocysteine (Hcys) levels by a 6-week methionine treatment produced remarkable glomerular injury. However, the mechanism by which hyperhomocysteinemia (hHcys) produces glomerular injury remains unknown. The present study was to observe when glomerular injury happens during hHcys and to explore the possible role of podocyte injury in the progression of glomerulosclerosis associated with hHcys. METHODS: Uninephrectomized Sprague-Dawley rats treated with methionine were used to examine the time course of glomerular injury induced by hHcys. RESULTS: Creatinine clearance was not different until rats were treated with methionine for 6 weeks, although plasma Hcys levels significantly increased at the 1st week of methionine treatment. However, urinary albumin excretion increased at the 2nd week of methionine treatment. Morphological examinations showed that mesangial expansion occurred at the 2nd week and podocyte effacement was also observed as processed glomerular damage during hHcys. Immunofluorescence analyses demonstrated that podocin and nephrin expressions were reduced, while alpha actinin-4 increased during hHcys. CONCLUSIONS: Increased plasma Hcys level is an important pathogenic factor resulting in glomerular injury even in the very early time of hHcys. These pathogenic effects of Hcys are associated with podocyte injury and changed expression and distribution of podocyte-associated proteins. PMID- 17396031 TI - Adverse drug events in cognitively impaired elderly patients. AB - BACKGROUND/AIMS: Adverse drug events (ADEs) are a frequent problem encountered in the elderly. The aim of this study was to elucidate the factors that influence ADEs in an elderly population with cognitive impairment. METHODS: 242 patients were recruited from dementia clinics and assessed after 6 months for ADEs. The use of natural health products (NHPs) was also documented. RESULTS: Backward logistic regression found that higher age (OR = 1.06; 95% CI 1.01-1.12), and greater cognitive impairment (OR = 0.94; 95% CI 0.90-0.98) were associated with an increased risk of developing an ADE while the use of NHPs (OR = 0.32; 95% CI 0.13-0.79) was associated with a decreased risk (chi(2) = 27.6, p < 0.001). CONCLUSION: Risk of ADEs increased with greater age and cognitive impairment but decreased with the use of NHPs. PMID- 17396032 TI - Association of extracellular cleavage of E-cadherin mediated by MMP-7 with HGF induced in vitro invasion in human stomach cancer cells. AB - BACKGROUND: Proteolytic shedding of the ectodomain of a variety of transmembrane proteins, including cell-to-cell adhesion molecules, has been observed in solid cancers. We have investigated whether extracellular cleavage of E-cadherin mediated by matrix metalloproteinase-7 (MMP-7) is involved in hepatocyte growth factor (HGF) induced in vitro invasion in stomach cancer cells. METHODS: The effects of HGF on the expression of E-cadherin/beta-catenin and MMP-7 at both the protein and mRNA levels were assessed in stomach cancer cells, NUGC-3 and MKN-28, and in cells in which the expression of MMP-7 was downregulated by transfection with a MMP-7 short hairpin RNA plasmid. RESULTS: Treatment with HGF increased the extracellular cleavage of E-cadherin and the release of MMP-7 and reduced the level of E-cadherin in a dose- and time-dependent manner. HGF treatment repressed the phosphorylation of beta-catenin in a Triton-soluble fraction, but enhanced this phosphorylation in a Triton-insoluble fraction. The association of E cadherin with beta-catenin was decreased by HGF treatment in the Triton-soluble fraction. In addition, treatment of MMP-7 short hairpin RNA transfected NUGC-3 cells with HGF resulted in no extracellular cleavage of E-cadherin and also decreased the in vitro cell invasion. CONCLUSIONS: These results suggest that incubation with HGF mediated the release of MMP-7, resulting in extracellular cleavage of E-cadherin from stomach cancer cells. This might be a key mechanism in HGF-induced in vitro invasion and metastasis. PMID- 17396033 TI - Cytokine-release patterns in elderly patients with systemic inflammatory response syndrome. AB - BACKGROUND: Information is scarce about the inflammatory responses to infection in elderly patients. OBJECTIVE: To investigate the inflammatory pattern in elderly patients with systemic inflammatory response syndrome (SIRS). METHODS: We assessed the value of pro-inflammatory cytokines (TNF-alpha, IL-6, IL-1beta) and C-reactive protein (CRP) in elderly patients with SIRS at the time of consultation in the emergency room and on the fourth day of hospitalization. RESULTS: Of 100 patients with SIRS, 92 had a clinical suspicion of infection and 46 were microbiologically confirmed. Basal TNF-alpha and IL-6 levels were higher in patients with bacteremia (p < 0.001 and p = 0.017, respectively). Basal levels of TNF-alpha (p < 0.001) and fourth-day levels of CRP (p = 0.004), TNF-alpha (p < 0.001) and IL-6 (p = 0.002) had a positive correlation with the length of hospitalization. Higher IL-6 levels on the fourth day showed a correlation with mortality (p = 0.016), while the other cytokines and CRP did not show this association. CONCLUSION: Febrile elderly patients showed an inflammatory response pattern to infection equivalent to that reported in younger patients. Despite some limitations of our study, among febrile elderly patients, pro-inflammatory cytokine levels can be good markers for evaluating the prognosis of infections. PMID- 17396034 TI - Final height in children with idiopathic growth hormone deficiency treated with a fixed dose of recombinant growth hormone. AB - There is no consensus regarding the optimal dosing of recombinant human growth hormone (rhGH) for children with growth hormone deficiency (GHD). Our objective was to evaluate the final adult height (FAH) in children with idiopathic GHD treated with a fixed rhGH dose of 0.18 mg/kg/week. We reviewed all charts of patients with idiopathic GHD treated with rhGH since 1985 who reached FAH. Ninety six patients were treated for an average of 5.4 years. The mean age was 11.9 years, the mean height -2.87 standard deviation score (SDS) and the mean FAH was 1.04 SDS. Females had a lower predicted adult height than males at the initiation of therapy (-2.0 vs. -1.01 SDS; p = 0.0087) but a higher FAH - predicted adult height (1.08 vs. 0.04 SDS; p = 0.0026). In multiple regression analysis, the FAH SDS was positively related to the midparental height SDS, the height SDS at GH initiation and growth velocity during the first year of therapy, and negatively correlated with peak GH and bone age at initiation (r(2) = 0.51; p < 0.005). Treatment of children with idiopathic GHD with a fixed dose of 0.18 mg/kg/week rhGH is sufficient to reach FAH within 2 SDS of the normal population range (84%) with an average FAH within -0.5 SDS of midparental height. PMID- 17396035 TI - Optimizing growth hormone therapy in growth hormone deficient children: what to do in the absence of hard evidence? Commentary on Rachmiel et al.: Final height in children with idiopathic growth hormone deficiency treated with a fixed dose of recombinant growth hormone (Horm Res 2007;68:236-243). PMID- 17396036 TI - Prevalence of Balkan endemic nephropathy has not changed since 1971 in the Kolubara region in Serbia. AB - BACKGROUND/AIMS: Thirty-one years after the first cross-sectional study, the population of Vreoci, a Balkan endemic nephropathy (BEN) village, was reinvestigated in order to determine the current prevalence of BEN and the clinical and laboratory characteristics of BEN family members with detected signs of kidney disease. METHODS: A total of 2,009 inhabitants (82% of the adult population) of the village were examined. Danilovic's criteria were used for diagnosis and classification of BEN. RESULTS: The prevalence of BEN (1.70%) was similar to that in 1971 (1.67%). Diagnosis of BEN was established in 19 BEN family members, suspected BEN in 23, proteinuria in 29, while 16 healthy members were examined as controls. Urine protein, alpha1-microglobulin levels and frequency of glucosuria were significantly higher and kidney length significantly smaller in the three patient groups than in healthy persons. Serum urea and creatinine levels were significantly higher, but creatinine clearance was lower in BEN and BEN suspected patients than in the other two groups. CONCLUSION: The prevalence of BEN remains stable over time in Vreoci village. Manifested disease was found in both BEN and BEN suspected patients. In persons with proteinuria but not enough criteria for BEN, tubular disorders and hypertension were frequently found. PMID- 17396037 TI - Treatment of lupus nephritis with cyclosporine - an outcome analysis. AB - BACKGROUND: The optimal therapy for lupus nephritis (LN), including the role of cyclosporine (CsA), still lacks scientifically valid clinical experience. We evaluated the efficacy of CsA in the induction and maintenance treatment of patients with biopsy-proven LN. PATIENTS AND METHODS: A total of 31 patients (25 women, 6 men, mean age 29.5 years) were enrolled in the study. The majority had proliferative LN. The mean follow-up was 85.6 +/- 24.7 months. RESULTS: CsA was used as first-line treatment in 38.7% of patients and as second-line treatment in 61.3% of patients. Complete remission was achieved in 93.5% of patients. The relapse rate was 45.2%. The mean disease-free interval was 33 months. At the end of follow-up, a total of 67.9% of the patients were in remission. The treatment led to significant improvement in proteinuria (p = 0.001) and stabilization of renal function. CONCLUSION: CsA might be an appropriate and a less toxic alternative drug for LN both as a first-choice and rescue therapy. PMID- 17396038 TI - Risk of persistent pulmonary hypertension of the neonate in twin-to-twin transfusion syndrome. AB - BACKGROUND: Chronic twin-to-twin transfusion syndrome (TTTS) is a complication of monochorionic twin gestations and is associated with high perinatal mortality and increased neurological, cardiovascular and renal morbidity. OBJECTIVE: To report the risk of severe persistent pulmonary hypertension of the newborn (PPHN) in TTTS and discuss the possible association between severe PPHN and TTTS. METHODS: All cases of monochorionic twins with severe PPHN at birth admitted to our nursery between June 2002 and July 2006 were reviewed retrospectively. We compared the incidence of severe PPHN in monochorionic twins with and without TTTS. Severe PPHN was diagnosed according to clinical and ultrasound criteria when an infant with a structurally normal heart had (1) severe hypoxemia and (2) evidence of a right-to-left shunt on persistent ductus arteriosus or foramen ovale, requiring treatment with inhaled nitric oxide (iNO). RESULTS: In a consecutive series of 73 twin pregnancies with TTTS, 4 of the 135 live-born twins (3%) were affected by severe PPHN. All reacted promptly to treatment with iNO. The incidence of severe PPHN in monochorionic twins without TTTS was 0% (0/161). CONCLUSION: In view of the severe clinical course in PPHN and need for adequate and prompt treatment with iNO, perinatologists should be aware of the increased risk of severe PPHN in TTTS. PMID- 17396039 TI - A randomised phase II study of irinotecan in combination with 5-FU/FA compared with irinotecan alone as second-line treatment of patients with metastatic colorectal carcinoma. AB - BACKGROUND: We conducted a randomised phase II study to compare irinotecan monotherapy with irinotecan in combination with infusional 5-fluorouracil/folinic acid (5-FU/FA) regarding efficacy and safety of these regimens in second-line therapy after failed fluoropyrimidine therapy in patients with metastatic colorectal cancer (mCRC). PATIENTS AND METHODS: 55 patients with mCRC after failure of a first-line therapy were randomised to receive either irinotecan 80 mg/m2 followed by FA 500 mg/m2 and 5-FU 2,000 mg/m2 24 h weekly for 6 weeks, with courses repeated on day 50 (arm A), or irinotecan 125 mg/m2 weekly for 4 weeks, with cycles repeated on day 43 (Arm B). RESULTS: Both regimens yielded a partial response rate of 11% with identical progression-free survival (3.7 months for both regimens) and similar overall survival (9.5 months for the combination therapy vs. 10.7 months for the monotherapy). Both regimens were very well tolerated, and the combination of irinotecan with 5-FU/FA did not result in increased toxicity. CONCLUSION: Our study confirms that irinotecan alone or in combination with infusional 5-FU/FA is an effective and safe regimen for CRC patients who failed first-line therapies. However, the role of 5-FU in addition to irinotecan for fluoropyrimidine failures remains unclear. Due to the small sample size, a decision cannot be made which therapy should be preferred, and a significant contribution to the efficacy of single-agent irinotecan is not obvious from this small randomised phase II trial. PMID- 17396040 TI - Effects of a step-by-step inpatient rehabilitation programme on quality of life in breast cancer patients. A prospective randomised study. AB - BACKGROUND: The aim of this study was to compare a conventional oncological inpatient rehabilitation programme with an inpatient step-by-step rehabilitation programme with regard to quality of life in breast cancer patients. PATIENTS AND METHODS: Women with the diagnosis of breast cancer were randomly assigned to either the step-by-step programme (group A: 3-week rehabilitation plus 4 and 8 months later a 1-week sojourn each time) or the conventional rehabilitation programme (group B: 4- week stay and no further interventions afterwards). Quality of life was assessed with the QLQ-C30 of the EORTC at t0 (beginning of the study), t1 (end of 3- or 4-week stay) and t2 (12 months after t0, end of study). RESULTS: Between t0 and t1, a tendency (0.1 > p = 0.05) could be shown in group B towards better improvement in global quality of life and emotional function. In the subgroup of patients that had impaired functions of quality of life at t0, superiority in group B reached significance (p < 0.05) for emotional and cognitive function at t1. Between t1 and t2, cognitive function improved further in group A, but diminished in group B. For the subgroup of patients with impaired cognitive function at t0, this difference between both groups became significant (p = 0.0098). CONCLUSIONS: Although not generally superior to conventional inpatient rehabilitation programmes, the analysed step-by- step rehabilitation provided marked benefits for patients with cognitive impairments. PMID- 17396041 TI - Safety and efficacy of itraconazole compared to amphotericin B as empirical antifungal therapy for neutropenic fever in patients with haematological malignancy. AB - BACKGROUND: Safety, tolerability and efficacy of itraconazole and amphotericin B (AMB) were compared for empirical antifungal treatment of febrile neutropenic cancer patients. PATIENTS AND METHODS: In an open, randomised study, 162 patients with at least 72 h of antimicrobial treatment received either intravenous followed by oral itraconazole suspension or intravenous AMB for a maximum of 28 days. Permanent discontinuation of study medication due to any adverse event was the primary safety parameter. Efficacy parameters included response and success rate for both treatment groups. RESULTS: Significantly fewer itraconazole patients discontinued treatment due to any adverse event (22.2 vs. 56.8% AMB; p < 0.0001). The main reason for discontinuation was a rise in serum creatinine (1.2% itraconazole vs. 23.5% AMB). Renal toxicity was significantly higher and more drug-related adverse events occurred in the AMB group. Intention-to-treat (ITT) analysis showed favourable efficacy for itraconazole: response and success rate were both significantly higher than for AMB (61.7 vs. 42% and 70.4 vs. 49.3%, both p < 0.0001). Treatment failure was markedly reduced in itraconazole patients (25.9 vs. 43.2%), largely due to the better tolerability. CONCLUSIONS: Itraconazole was tolerated significantly better than conventional AMB and also showed advantages regarding efficacy. This study confirms the role of itraconazole as a useful and safe agent in empirical antifungal therapy of febrile neutropenic cancer patients. PMID- 17396042 TI - Complete remission and early relapse of refractory plasma cell leukemia after bortezomib induction and consolidation by HLA-mismatched unrelated allogeneic stem cell transplantation. AB - BACKGROUND: Chromosomal deletion of q13 (del q13) and plasma cell leukemia predict both a worse prognosis in myeloma. Experiences with bortezomib in plasma cell leukemia prior to allogeneic stem cell transplantation (SCT) have not yet been reported. CASE REPORT: A 66- year-old male patient was admitted for IgA myeloma(IIIA) with del q13. The myeloma progressed to plasma cell leukemia. Bortezomib was given, free light chain (FLC) excretion decreased, and myeloma cells disappeared from blood and decreased in marrow. An unrelated, mismatched allogeneic SCT was performed. The patient was discharged after engraftment with full chimerism without signs of GvHD. FLC excretion increased, and immunosuppression was discontinued. From day +84 on, bortezomib was infused again and FLC excretion decreased rapidly. Relapse was confirmed in marrow, and bortezomib was continued. Donor lymphocyte infusions (DLI) had no effect, and a further cycle of bortezomib and thalidomide had only minor effects. On day +209, the patient died from myeloma. CONCLUSION: This case gives evidence for an excellent initial response of plasma cell leukemia to bortezomib, however, early relapse after SCT clearly indicates limitations of both bortezomib therapy and allogeneic SCT in high-risk myeloma. Future developments are mandatory and could include combination of bortezomib with other cytostatics and early DLI in the absence of GvHD. In addition, there is need to clarify if graft-versus-myeloma effect is diminished by bortezomib therapy after allogeneic SCT. PMID- 17396043 TI - Recurrent low- to intermediate-grade chondrosarcoma of the thumb with lung metastases: an objective response to trofosfamide. AB - BACKGROUND: Low- to intermediate-grade chondrosarcoma is usually a slow-growing and highly chemotherapyresistant tumor type. CASE REPORT: The 76-year-old female patient presented with low- to intermediate-grade chondrosarcoma of the distal phalanx of the right thumb in 1993 and was treated with an excision of the carpometacarpal joint. Approximately 10 years later, the patient presented with recurrent local disease at the trapezoid bone, which was resected. Eight months later she presented with local recurrence at the carpal joint and lung metastases. Subsequently she was treated chemotherapeutically with pegylated liposomal doxorubicin as part of a clinical trial. Due to tumor progression after 2 cycles, she was switched to oral trofosfamide (150 mg daily). A follow-up CT scan after 8 months demonstrated a partial remission that was confirmed by a second scan after 9 months of treatment. The treatment was well tolerated. After 18 months of continued treatment, she was still in remission and on trofosfamide. CONCLUSION: Trofosfamide at a daily dose of 150 mg may be effective in low- to intermediate-grade chondrosarcoma. PMID- 17396044 TI - Asymptomatic primary malignant melanoma of the esophagus. AB - BACKGROUND: Primary malignant melanoma of the esophagus is an exceedingly rare disease. This tumor is typically aggressive and disseminates early via the lymphatics and the bloodstream with a mean survival time between 10 and 15 months after radical surgical resection. The role of chemotherapy and immunotherapy is unclear. No treatment plan for the disease has yet been established. CASE REPORT: A 78-year-old man came for a checkup with a medical history of reflux esophagitis and chronic gastritis. Esophagogastroscopy showed a bluishgray tumor of the esophagus, and histology revealed features consistent with malignant melanoma. The patient underwent total transhiatal esophagectomy with curative intention, and esophagogastric anastomosis was performed. Immunohistochemistry revealed tumor cells strongly positive for the melanoma-specific antigen HMB45 and protein S-100, and negative for cytokeratin. A proposed postoperative chemotherapy was declined by the patient. Nine months after surgery, the patient's condition deteriorated, and a mediastinal lymph node conglomerate was found. Two months later, he died of bleeding into the cervical soft tissue. CONCLUSION: Up to date, radical surgical resection is the main treatment. Very little is known about the benefits of chemotherapy and immunotherapy. However, these therapeutic modalities may play an important role in the future. PMID- 17396045 TI - An ambiguous phenomenon of radiation and drugs: recall reactions. AB - The term 'radiation recall' describes an acute inflammatory reaction in previously irradiated areas after the administration of certain inciting systemic agents. It was first described in 1959 by D'Angio that dermatitis is related to the application of actinomycin D on the skin. Though this reaction occurs frequently on the skin, it may also be seen in the oral mucosa, the larynx, esophagus, small intestine, lungs, muscle tissue, and brain. Most drugs associated with recall reactions are cytotoxics, however, several other drugs may also elicit the phenomenon. Although this phenomenon is well known, its etiology is not understood. Radiation recall reactions are generally associated with megavoltage radiotherapy. The time interval between the completion of radiotherapy and the recall reaction ranges from days to years. The recall reaction occurs on average 8 days (3 days to 2 months) after the application of the promoting agent. Although no standard treatment exists, some authors suggest discontinuation of the inciting drug and the use of corticosteroids or nonsteroidal anti-inflammatory agents. PMID- 17396046 TI - Fibromyalgia, facial expression and emotional experience. AB - BACKGROUND: The present study aims at analyzing the nonverbal affective behavior of female fibromyalgia (FM) inpatients in comparison to healthy women. METHODS: Videotaped psychodynamic interviews of each of 15 female FM inpatients and healthy women were analyzed. Afterwards the analyses of facial expression were related to gazing behavior and emotional experience. RESULTS: FM patients exhibited neither a reduction in total activity of facial expression nor in absolute frequency of primary affects in comparison to healthy women, who, however, (also in eye contact) also exhibited a significantly higher proportion of 'genuine joy' and a lower one of 'contempt'. No congruence between the patient's emotional experience and affective expression was found. CONCLUSIONS: The absence of reduced total activity of facial expression is in contrast to the elaborate descriptions of complaints provided by the patients. Nevertheless, our detailed analysis shows a lack of elements that stabilize the relationship and the presence of dissociating elements in the interactions. PMID- 17396047 TI - The operationalized psychodynamic diagnostics system: clinical relevance, reliability and validity. AB - In this paper, we present a multiaxial system for psychodynamic diagnosis, which has attained wide usage in Germany in the last 10 years. First we will discuss the 4 operationalized psychodynamic diagnostics (OPD) axes: illness experience and treatment assumptions, relationships, mental conflicts, and structure, then clinical applications will be outlined. Focus psychodynamic formulations can be employed both with inpatients and with outpatients. Studies show good reliability in a research context and acceptable reliability for clinical purposes. Validity will be separately summarized as content, criterion, and construct validity. Validity studies indicate good validity for the individual axes. Numerous studies on the OPD indicate areas of possible improvement, for example for clinical purposes the OPD should be more practically formulated. PMID- 17396048 TI - Intentionality as a link between the neuropsychology and the symptoms of schizophrenia. AB - Cognitive deficits are an important feature of schizophrenia, particularly in the areas of working memory and executive function. However, their relationship to other clinical dimensions of the illness has remained elusive. We suggest that a phenomenological exploration of disordered intentionality in schizophrenia symptoms might provide us with a framework for understanding the role of cognitive deficits. We propose an understanding of intentionality as having a layered structure, which facilitates a mapping onto neuropsychological concepts. A disturbance of basic intentional processes can explain some of the 'positive' symptoms of schizophrenia. We focus on the temporal aspect of intentionality, which will be related to recent conceptions of prefrontal cortical function as the temporal organization of thought and behavior. A compensation of these basic disturbances leads to a 'minimization of change', which can explain some of the 'negative' symptoms. The compensatory strategies are thought to rely on the higher-order executive functions of the prefrontal cortex. These are also disturbed in schizophrenia, which often renders the process of compensation ineffective, leading to a fragmentation of thought and behavior. Thus the concept of intentionality might help us relate different aspects of prefrontal dysfunction to specific schizophrenic symptom clusters. PMID- 17396049 TI - The temporal structure of intentionality and its disturbance in schizophrenia. AB - Working memory, attention and executive control functions are central areas of neuropsychological research in schizophrenia. These concepts implicitly refer to the basic temporal structure of mental life as an integration of past, present and future. From a phenomenological point of view, they may be paralleled to the structure of internal time consciousness as analyzed by Husserl, consisting of a retentional, presentational and protentional function. These synthetic functions, operating at the most basic layer of consciousness, are capable of integrating the sequence of single moments into an 'intentional arc', enabling us to direct ourselves towards objects and goals in a meaningful way. On this background, basic symptoms of schizophrenia such as formal thought disorder, loss of automatic performances and disturbances of self-awareness may be conceived as caused by a weakening and dissolution of the intentional arc. A failure of the continuous intertwining of succeeding moments, and especially of the protentional function, leads to a loss of the tacit or operative intentionality that carries the acts of perceiving, thinking and acting. The loss of tacit, implicit functions undermines the common-sensical understanding of reality and has to be compensated by the deliberate, hyperreflexive reconstruction of everyday performances. Phenomenological analyses may thus establish a link between experimental research on single mental dysfunctions on the one hand and the higher level of the patient's subjective experience on the other. PMID- 17396050 TI - The differentiation between 'lack of insight' and 'dysfunctional health beliefs' in schizophrenia. AB - BACKGROUND: 'Lack of insight' into one's own illness is a frequent symptom in schizophrenic disorders. 'Health beliefs' are psychological explanations of one's own health status and are held by all individuals. The question is how they relate to each other. METHODS: Lack of insight (according to the definition of the AMDP System) and health beliefs (measured with the Illness Concept Scale) were assessed in 364 schizophrenic outpatients who participated in a study on neuroleptic long-term treatment. RESULTS: 'Insight into illness' and 'health beliefs' are independent of each other. Insight is related to the current psychopathological status as measured by the Brief Psychiatric Rating Scale, whereas health beliefs are related to personal life experiences. Lack of insight and dysfunctional health beliefs are both associated with patient noncompliance. CONCLUSION: Poor insight and dysfunctional health beliefs are separate clinical phenomena. Making this distinction is important for guiding research and patient care. PMID- 17396051 TI - Gender differences of enzymatic activity and distribution of 17beta hydroxysteroid dehydrogenase in human skin in vitro. AB - The interconversion of estrone (E1) and 17beta-estradiol (E2) is catalyzed by 17beta-hydroxysteroid dehydrogenase (17beta-HSD) in peripheral steroidogenic organs such as the skin. To investigate gender differences of activity and skin distribution of 17beta-HSD in human skin, enzymatic activity was measured in skin homogenates and skin horizontally sliced by 10 microm thickness in vitro. Reductive 17beta-HSD (E2 formation from E1) in female skin has a lower substrate affinity than in male skin; Km (Michaelis-Menten constant) of female and male skin is 11.8 +/- 6.5 and 2.0 +/- 2.0 microM, respectively. Female skin had a tendency to activate estrogen; Vmax (maximum rate) for E2 formation, 5.8 +/- 4.0 pmol/min/mg protein, is 1.7 times larger than E1 formation, 3.5 +/- 1.5 pmol/min/mg protein, and, on the other hand, male skin tends to deactivate estrogen; Vmax for E1 and E2 is 10.5 +/- 6.1 and 4.2 +/- 3.7 pmol/min/mg protein, respectively. The concentration of metabolite had a peak value at 80-120 microm from the skin surface. Therefore, these in vitro results suggest that the enzymatic activities of 17beta-HSDs have a gender difference in estrogen formation/metabolism and are distributed around the basement layer of the epidermis irrespective of sex. 17Beta-HSDs distributed around the basement epidermis may be effectively supplied with circulating estrogen from the papillary plexus to maintain the estrogen level in skin. This distribution pattern having a peak surrounding 100 microm from the skin surface indicates the importance for defense from noxae (e.g. detoxication) and maintenance of the internal environment (e.g. biosynthesis of hormones). Future studies should increase sample size and confirm these results by stricter statistical analysis. PMID- 17396052 TI - Interaction of lipophilic moisturizers on stratum corneum lipid domains in vitro and in vivo. AB - Dry skin symptoms such as scaling and itching are often treated with lipophilic moisturizers. The aim of this study was to investigate the effect of lipophilic moisturizers on the stratum corneum (SC) ultra-structure and lipid organization. Lipophilic moisturizers were applied on the forearms of 4 healthy volunteers for 3 h. Subsequently, the application sites were tape stripped, and selected tape strips prepared for Freeze Fracture Electron Microscopy (FFEM), a method to visualize the SC intercellular lipid parallel to the skin surface. To investigate the effect of lipid moisturizers on the lipid lamellae, isolated SC was pretreated with the lipophilic moisturizers for 24 h prior to performing small angle X-ray diffraction (SAXD) measurements. Additionally, the lipid organization of mixtures prepared with ceramides, cholesterol, free fatty acids and lipophilic moisturizer in a 2:1:1:1 molar ratio were studied using SAXD. The FFEM data (in vivo) as well as the SAXD data (in vitro) show that the lipophilic moisturizers do not change the lipid lamellar organization in the SC. Addition of 20% m/m lipophilic moisturizer to the ceramide:cholesterol:free fatty acids mixture did not inhibit the formation of the long periodicity phase, the characteristic lamellar phase in the SC, even though there was clear evidence that two of the three moisturizers were at least partially incorporated in the long periodicity phase. Concluding, all findings suggest that the lipophilic moisturizers investigated in this study do not drastically change the lamellar organization of the SC intracellular lipid matrix, but that the moisturizers form separate domains in the SC, as was visualized by FFEM. PMID- 17396053 TI - Kinetic characteristics of acidic and alkaline ceramidase in human epidermis. AB - It has recently become evident that at least five ceramidase (CDase) isoforms are present in human epidermis, and that specifically acidic CDase (aCDase) and alkaline CDase (alkCDase) activities increase during keratinocyte differentiation, and thus might play a pivotal role(s) in permeability barrier function. Prior to investigating their possible roles in the epidermal barrier function, it is necessary to characterize basic kinetic parameters for these enzymes, as well as to determine the effects of the established CDase inhibitors and their activities. In this study, assays for both aCDase and alkCDase activities in fully differentiated human epidermis were optimized using a radiolabeled substrate. These studies revealed that aCDase activity is substantially higher than alkCDase activity, and that both isoenzymes are inhibited by a CDase inhibitor N-oleylethanolamine. These findings were also confirmed using an in situ enzyme assay. PMID- 17396054 TI - Follicular penetration of topically applied caffeine via a shampoo formulation. AB - AIMS: Follicular drug delivery is the prerequisite for an effective treatment of androgenetic alopecia or other reasons of premature hair loss. METHODS: The follicular penetration of caffeine, applied topically in a shampoo formulation for 2 min, was measured with highly sensitive surface ionization in combination with mass spectroscopy, a selective method for the detection of very small quantities of transcutaneously absorbed substances in the blood. An experimental protocol, developed to selectively block the follicular pathway within the test area, was used. Based on this principle, a clear distinction between interfollicular and follicular penetration of topically applied caffeine was feasible. RESULTS: After 2 min, caffeine penetrated via the hair follicles and stratum corneum. CONCLUSION: It was found that the penetration via hair follicles was faster and higher compared with the interfollicular route and that hair follicles are the only pathway for fast caffeine absorption during the first 20 min after application. PMID- 17396055 TI - Blunted hemodynamic response and reduced oxygen delivery with exercise in anemic heart failure patients with systolic dysfunction. AB - Anemic heart failure patients with systolic dysfunction are known to have reduced exercise capacity. Whether this is related to poor hemodynamic adaptation to anemia is not known. Peak exercise oxygen consumption (VO2) and hemodynamics at rest and peak exercise were assessed among 209 patients and compared among those who were (n=90) and were not (n=119) anemic. Peak VO2 was significantly lower among anemic patients (11.7+/-3.3 mL/min/kg vs 13.4+/-3.1 mL/min/kg; P=.01). At rest, right atrial pressure was higher (10+/-5 mm Hg vs 8+/-4 mm Hg; P=.02) and venous oxygen saturation lower (62%+/-8% vs 58%+/-10%; P<.01) among anemic patients. At peak exercise, anemic patients had a higher wedge pressure (27+/-9 mm Hg vs 24+/-10 mm Hg; P=.04). No significant differences in stroke volume, cardiac index, systemic vascular resistance, or oxygen saturation were noted between the 2 groups. In conclusion, the relative hemodynamic response to exercise among anemic heart failure patients appears blunted and may contribute to worse exercise tolerance. PMID- 17396056 TI - The effects of obesity on functional work capacity and quality of life in phase II cardiac rehabilitation. AB - Many patients referred to cardiac rehabilitation (CR) programs are obese. It is unknown, however, whether obese patients derive reduced benefit from CR. A total of 388 CR patients were categorized into normal-weight, overweight, class I obese, and class II/III obese groups. Functional work capacity and quality of life were examined at baseline and after 12-week completion of the CR program. After adjusting for demographic and medical conditions, class II/III obese persons showed lower work capacity and physical quality of life at both baseline and follow-up. Class II/III obese individuals also showed smaller gains in work capacity from baseline to follow-up than all other groups. Further work is needed to identify strategies for improving outcome in obese patients, including incorporating structured weight loss into CR or post-CR referral to an exercise maintenance program. PMID- 17396057 TI - Variation in blood levels of inflammatory markers related and unrelated to smoking cessation in women. AB - This study assessed the influence of short-term changes in smoking habit on blood levels of inflammatory markers, which have been associated with increased cardiovascular risk. Five inflammatory markers were measured before and 6 weeks after attempting smoking cessation in 138 healthy women. In the 48 participants who stopped smoking, white blood cell count (-0.7+/-1.2 x 10(9)/L; P<.001) and fibrinogen (-0.6+/-1.5 micromol/L; P<.01) decreased, but there was no significant (P>.1) change in the plasma level of C-reactive protein (median change +0.1; interquartile range -0.2, 0.9 mg/L), intercellular adhesion molecule 1 (+17+/-75 ng/mL), or CD40 ligand (+0.4+/-2.1 ng/mL). Most of the individual variation in inflammatory marker levels was unrelated to changes in smoking habit. PMID- 17396058 TI - Are emergency chest pain patients ready to quit smoking? AB - This study examined predictors of readiness to quit smoking among emergency chest pain patients admitted to the observation unit (OU) to rule out myocardial infarction. While in the OU, patients (n=543) completed surveys assessing smoking history, nicotine dependence, readiness to quit, and other relevant variables. Participants smoked an average of 18.8 (SD=12.6) cigarettes per day. More than half (58%) had made at least 1 serious quit attempt > or = 24 hours) in the past year. Most had never used nicotine replacement medications. Nicotine dependence, perceived risk from smoking, and patient perceptions that smoking might be related to their chest pain were significantly associated with readiness to quit (P<.05). Results indicate that a significant proportion of OU patients think they are at relatively low risk from smoking and, although motivated to quit, are not using medications appropriately to assist quit attempts. There is a need for intervention and education with this population of patients. PMID- 17396059 TI - Effects of levels of cigarette smoke exposure on symptom-limited spiroergometry. AB - Previous investigations demonstrated reduced exposure to selected cigarette smoke constituents in adult smokers switching from conventional cigarettes (CC) to an electrically heated cigarette smoking system (EHCSS). This study investigated whether reduced exposure and no smoking (NS) would improve exercise performance. In a 3-period crossover study, 18 male adult smokers (age, 43.6+/-5.3 years) of CC were randomized to smoke CC (tar, 11 mg; nicotine, 0.8 mg; carbon monoxide, 11 mg), to use EHCSS (tar, 3 mg; nicotine, 0.2 mg; carbon monoxide, 0.4 mg [Federal Trade Commission method]), or to NS for 3 days before performing symptom-limited spiroergometry. NS and EHCSS vs CC resulted in less severe dyspnea (NS, 44.4% [P<.01 vs CC;] EHCSS, 50% [P=.03 vs CC;] CC, 88.9%), higher working capacity (NS, 2.92+/-0.4 W/kg [P=.06 vs CC;] ECHSS, 2.92+/-0.4 W/kg [P=.04 vs CC;] CC, 2.86+/ 0.5 W/kg), higher peak oxygen uptake (NS, 2694+/-466 mL O(2)/min [P=.08 vs CC;] EHCSS, 2830+/-606 mL O(2)/min [P=.03 vs CC;] CC, 2682+/-492 mL O(2)/min), higher anaerobic threshold (NS, 1324+/-306 mL O(2)/min; EHCSS, 1396+/-312 mL O(2)/min [P=.03 vs CC;] CC, 1315+/-290 mL O(2)/min), and higher maximum rate-pressure product (NS, 30.1+/-2.7 x 10(3) mm Hg/min; EHCSS, 2.8 x 10(3) mm Hg/min [P<.01 vs CC;] CC, 30.7+/-29.2+/-3.6 x 10(3) mm Hg/min) indicating that reduced exposure from tobacco smoke and NS for 3 days may improve cardiovascular function as detected by symptom-limited spiroergometry. PMID- 17396060 TI - Stress testing in patients with implantable cardioverter-defibrillators: a preliminary report. AB - This retrospective study was undertaken to assess the responses to, and complications associated with, stress testing in patients with implantable cardioverter-defibrillators (ICDs). Primary end points were occurrence of malignant ventricular arrhythmias, onset of burst pacing or ICD firing, cardiopulmonary resuscitation, or death during or soon after stress testing. Secondary end points were urgent coronary revascularization and/or hospital readmission for cardiovascular complications. During a 4-year period, 1734 patients underwent ICD implantation or generator replacement at our institution; 84 patients (mean age +/- SD, 67+/-12 years; 76% men) subsequently underwent 107 stress tests, including 44 exercise and 63 pharmacologic (22 dobutamine, 41 dipyridamole) evaluations. None of the ICDs were inactivated before testing. All tests were supervised by specially trained paramedical personnel, with a physician immediately available. Four patients had self-terminating, nonsustained ventricular tachycardia at peak stress. None had sustained ventricular tachycardia requiring emergent therapy. There were no deaths or hospital readmissions for ventricular arrhythmias. These findings suggest that stress testing is feasible in patients with ICDs and that it can be performed without pretest inactivation. PMID- 17396061 TI - Smoking-related morbidity and mortality in the cardiovascular setting. AB - It is estimated that during the 20th century, 100 million people died of tobacco related illnesses worldwide. The outlook for the future is grim as this number is expected to increase 10-fold during the 21st century. Many of these deaths will occur in middle-aged adults and, while most tobacco-related deaths currently occur in men, female mortality is expected to increase markedly due to increased rates of smoking in women, especially in developing countries. The risk of coronary heart disease is strongly associated with smoking in both developed and undeveloped countries. In addition, other forms of tobacco exposure (chewing, inhalation through water, and secondhand smoke) have also been documented to be important causes of coronary disease worldwide. Fortunately, the news is not all bad. Recent large-scale studies show that much of the excess risk associated with smoking is attenuated 1 to 2 years after quitting, depending on the level of smoking during an individual's lifetime. These latest findings should stimulate efforts of health care workers to become more aggressive toward cessation of smoking in our practices. PMID- 17396062 TI - A clinical imperative: assisting patients who smoke to reduce their risk of cardiovascular disease. AB - The role of tobacco smoking as a cause of cardiovascular disease is now unequivocal and well-documented in literally hundreds of epidemiologic and biomedical studies over the past 50 years. Cessation of smoking, on the other hand, swiftly and profoundly reduces the risks of a cardiovascular event. Thus, smoking cessation should be seen as perhaps the most effective lifesaving intervention in the physician's armamentarium. Despite this widely available knowledge, and evidence that most smokers want to quit, relatively few physicians offer cessation support to their smoking patients, even those at high risk for a cardiovascular event. This article reviews the links between tobacco smoking and cardiovascular disease, argues for a greater role for physicians in assisting smokers to quit, and highlights the most effective interventions currently available. PMID- 17396063 TI - Smoking cessation pharmacotherapy--nicotine and non-nicotine preparations. AB - Cigarette smokers tend to die prematurely from a number of diseases. Cigarette smoking is an important modifiable risk factor for cardiovascular morbidity and mortality. Despite the clear health benefits of smoking cessation, smokers usually find it difficult to stop and behavioral therapies often prove insufficient. Pharmacologic intervention may aid the process because of the addictive nature of nicotine. Nicotine replacement therapy, which is regarded as first-line therapy, was developed to overcome the symptoms of nicotine withdrawal that many patients find distressing. Different modes of administration include inhalation and buccal or transdermal absorption. The orally administered non nicotine drugs varenicline and bupropion are also regarded as first-line treatments, either used alone or as an adjunct to nicotine replacement therapy. Second-line treatments include clonidine and nortriptyline. Other treatment strategies that have been examined include monoamine oxidase inhibitors and selective serotonin reuptake inhibitors; efficacy has yet to be proven definitively. A novel approach to treatment using the cannabinoid-1 receptor antagonist rimonabant is also under investigation. PMID- 17396064 TI - Maximizing smoking cessation in clinical practice: pharmacologic and behavioral interventions. AB - Clinicians are in a unique position to reduce cardiovascular morbidity and mortality by helping their patients quit smoking. At each visit, clinicians should document smoking status, provide strong and clear advice to quit, and recommend and prescribe pharmacotherapy for patients who are motivated to quit, which can double the odds of success. Effective pharmacotherapies include nicotine replacement, bupropion, and varenicline, which was recently approved by the Food and Drug Administration. Behavioral counseling to develop a quit plan and extended follow-up are critical to maximize quit rates but are rarely provided by clinicians due to time constraints and lack of expertise. As an alternative, the authors recommend referral to telephone quitlines that provide no-cost behavioral counseling by specialists. Hospitals should implement systemwide procedures to ensure that smokers are identified at admission and trained staff is available to provide smoking cessation consults that include a minimum of 20 minutes of inpatient counseling with follow-up for at least 1 month. PMID- 17396065 TI - Vascular disease in an expanding world: seeking answers inside and outside of the box. PMID- 17396067 TI - Effects of warfarin on blood pressure in men with diabetes and hypertension--a longitudinal study. AB - Warfarin causes extensive vascular calcification leading to increased systolic blood pressure and pulse pressure in rats, may be associated with increased valvular and coronary calcifications in man, and possibly worsens hypertension in high-risk patients, particularly in those with diabetes mellitus or uncontrolled hypertension. The authors evaluated blood pressure and intensity of antihypertensive therapy over 36 months in a cohort of 58 patients with diabetes and hypertension on warfarin and 58 control subjects with diabetes and hypertension not on warfarin. The results demonstrate that warfarin therapy at conventional doses does not increase systolic blood pressure or pulse pressure in patients with diabetes and hypertension. PMID- 17396066 TI - Effect of combined treatment with alpha-Lipoic acid and acetyl-L-carnitine on vascular function and blood pressure in patients with coronary artery disease. AB - Mitochondria produce reactive oxygen species that may contribute to vascular dysfunction. alpha-Lipoic acid and acetyl-L-carnitine reduce oxidative stress and improve mitochondrial function. In a double-blind crossover study, the authors examined the effects of combined alpha-lipoic acid/acetyl-L-carnitine treatment and placebo (8 weeks per treatment) on vasodilator function and blood pressure in 36 subjects with coronary artery disease. Active treatment increased brachial artery diameter by 2.3% (P=.008), consistent with reduced arterial tone. Active treatment tended to decrease systolic blood pressure for the whole group (P=.07) and had a significant effect in the subgroup with blood pressure above the median (151+/-20 to 142+/-18 mm Hg; P=.03) and in the subgroup with the metabolic syndrome (139+/-21 to 130+/-18 mm Hg; P=.03). Thus, mitochondrial dysfunction may contribute to the regulation of blood pressure and vascular tone. Further studies are needed to confirm these findings and determine the clinical utility of alpha lipoic acid/acetyl-L-carnitine as antihypertensive therapy. PMID- 17396068 TI - Pharmacologic agents in the management of hypertension--nisoldipine coat-core. AB - Nisoldipine coat-core (CC), a 1,4-dihydropyridine calcium antagonist, is indicated for the treatment of hypertension and may be used alone or in combination with other antihypertensive agents. The CC technology allows for extended delivery of the drug and once-daily dosing. Nisoldipine CC tablets are absorbed across the entire gastrointestinal tract, including the colon. Eighty percent of the total dose is in the slow-release outer coat, while the core has immediate-release characteristics suitable for absorption in the distal gastrointestinal tract. Numerous double-blind, randomized studies of this agent have been done in patients with hypertension. The use of nisoldipine CC reduced both clinic and ambulatory blood pressure to a similar degree when compared with angiotensin-converting enzyme (ACE) inhibitors, beta-blockers, and the calcium antagonists amlodipine and felodipine. The drug has also been studied in hypertensive African Americans and demonstrated equivalent efficacy to amlodipine. Tolerability of the drug is good, with the most common side effect of edema at a rate similar to other dihydropyridine calcium antagonists. Thus, results of more than a decade of clinical trial data support the use of nisoldipine CC as once-daily therapy for the treatment of hypertension. PMID- 17396069 TI - Automated measurement of blood pressure in routine clinical practice. AB - In recent years, automated devices have been developed to record blood pressure (BP) accurately in the home and during usual daily activities. Clinical outcome studies have clearly shown home BP and 24-hour ambulatory BP to be significantly better predictors of future cardiovascular events compared with BP recorded in the office setting using mercury sphygmomanometry. It is also now possible to measure office BP with the patient resting quietly alone in the examining room using an automated device. Studies in routine clinical practice using this approach have demonstrated that automated office BP can eliminate most of the white coat effect seen with manual BP measurement. The automated office BP also correlates significantly better than does the routine office BP with the 24-hour ambulatory BP, the gold standard for predicting risk of future cardiovascular events. Sufficient evidence now exists to consider incorporating automated office BP into an algorithm for diagnosing hypertension. PMID- 17396070 TI - Adherence. PMID- 17396071 TI - First-step use of fixed-dose combination treatment in stage 2 hypertensive patients: has the time come? PMID- 17396072 TI - More light please. PMID- 17396073 TI - Hepatocellular necrosis associated with labetalol. PMID- 17396074 TI - Exaggerated systolic blood pressure response to exercise. PMID- 17396075 TI - Is it a pheochromocytoma? AB - The patient is a 44-year-old man with a 4-year history of intermittently elevated blood pressure (BP) controlled by diet and exercise. Three months before evaluation he described daily "spikes" of BP with sharp unilateral headaches. He was seen in the emergency department with a BP of 212/106 mm Hg and was started on hydrochlorothiazide 25 mg daily. He denied palpitations, diaphoretic episodes, pallor, and tremor. The patient did not want to take medication and specifically requested an evaluation to rule out pheochromocytoma. Results from 24-hour urine tests for total metanephrines was 812 mg/24 h (normal, 130-520 mg/24 h), for total catecholamines was 53 mg/24 h (normal, 0-135 mg/24 h), and for vanillylmandelic acid was 4.7 mg/24 h (normal, <7 mg/24 h). Thyroid-stimulating hormone was 0.87 (normal, 0.4-4.0 IU/mL). Physical examination revealed normal optic fundi, negative cardiac examination results, and presence of peripheral pulses without bruits. His BP was now 136/74 mm Hg, with a heart rate of 76 beats per minute. PMID- 17396081 TI - [Endocannabinoids: therapeutic perspectives in chronic liver diseases]. PMID- 17396082 TI - Epidemiology of hepatocellular carcinoma in Finistere. Prospective study from June 2002 to May 2003. AB - OBJECTIVES: The aims of this prospective study were to evaluate the incidence of hepatocellular carcinoma (HCC) in Finistere, an administrative district of western France, and to highlight epidemiological characteristics. METHODS: From June 1st 2002 to May 31st 2003, all cases of HCC in Finistere were registered prospectively. Standardized incidence rates were calculated. RESULTS: One hundred and six cases of HCC were registered. Standardized incidence rates were 13.8/100000 among men and 0.8/100000 among women. Among the incident cases of HCC, 52% were discovered in patients consulting for symptoms, and 28% in patients undergoing screening. The new non-invasive diagnostic criteria of HCC were used in 60% of cases. HCC was associated with cirrhosis in 89 patients (84%). Excessive alcohol intake was the main cause of cirrhosis. In all, 27 patients (25.5%) received potentially curative treatment. CONCLUSION: Incidence of HCC is high in Finistere. The proportion of patients given potentially curative treatment is still low, illustrating how difficult screening can be when alcoholic cirrhosis predominates in the target population. PMID- 17396083 TI - [Carbohydrate metabolism dysregulation in cirrhosis: pathophysiology, prognostic impact and therapeutic implications]. AB - The liver plays a key-role in carbohydrates metabolism. Glucose intole-rance, overt diabetes mellitus and insulin resistance are characteristic features of patients with cirrhosis. Central hyperinsulinemia and peripheral insulin resistance are the main explanations for the high prevalence of diabetes in patients with cirrhosis. On the other hand, type 2 diabetes is associated with a wide spectrum of liver diseases ranging from nonalcoholic fatty liver to cirrhosis and hepatocellular carcinoma. Carbohydrate metabolism abnormalities are a major aggravating risk factor in cirrhosis. Diabetes is also an independent negative prognostic factor in cirrhotic patients. This leads to specific diagnostic procedures and therapeutic issues. Patients with diabetes and liver disease frequently need insulin treatment. The presence of liver disease makes the treatment of diabetes complex, and additional research is needed to determine the best treatment strategies in these patients. PMID- 17396084 TI - Patient information for liver biopsy: impact of a video movie. AB - AIMS: The information given to patients before a medical procedure is usually delivered verbally and in the form of a written document. Viewing a video movie about the procedure might be helpful in improving the quality of patient information and thus contributed to reduce patient anxiety. The aim of this prospective study on the methods used to deliver information to patients scheduled for liver biopsy was to determine, in comparison with standard information delivery, the impact on anxiety and understanding of additional information provided by a video movie. METHODS: Patients included in this study were scheduled for liver biopsy. Three days before the procedure, a physician informed the patients about liver biopsy and gave them a written information document. After this standard information delivery, the patients were randomly assigned to two groups, to view or not a video movie illustrating liver biopsy and recalling the information provided in the information document. Anxiety was measured after the information visit and just before the biopsy procedure using Spielberger's self-administered STAI-Y questionnaire. Patient understanding of and satisfaction with the information received were evaluated respectively before and after the procedure. RESULTS: Among the 67 patients retained for analysis, 33 viewed the information video and 34 did not. The level of anxiety measured at the information visit and before liver biopsy was similar in the two groups (38.8 and 37.4 with video versus 38.9 and 40.1 without video). The patients had an excellent understanding of the information received (12.3/14 with video; 12.7/14 without video) and were well satisfied with the information received. CONCLUSION: The patients understood and were well satisfied with the information received about liver biopsy. Complementary information in the form of a video movie had no effect on patient anxiety. PMID- 17396085 TI - [Sentinel node biopsy in colorectal cancer. A multifunctional tool or a missing link?]. PMID- 17396086 TI - Sentinel lymph nodes of colorectal carcinoma: reappraisal of 123 cases. AB - AIMS: Results concerning the usefulness of the sentinel lymph node (SLN) in colorectal carcinoma have been discordant. The SLN technique may be used to guide surgical resection (lymph mapping), restrict the lymph node analysis solely to the SLN (accuracy) and upgrade tumor staging when micrometastases are specifically detected in the SLN. METHODS: The blue dye injection technique was used. Serial sections of the SLNs were analyzed after hematoxylin-eosin (HES) staining. RESULTS: The SLN technique was tested in 123 patients, successfully in 112/118 (feasibility 95%) (five intraoperative exclusions). On average, twenty lymph nodes (range: 5-74) and two SLNs (range: 1-5) were identified. Lymph mapping was used in 11% of patients to guide surgical resection; the SLN was negative in 14 of 36 N+ patients (39% false-negatives); HES staining enabled detection of micrometastases in 8 of 84 initially N0 patients (10% secondary upgrading to N+). CONCLUSION: Limiting node analysis to the SLN cannot replace a complete pathology examination of all resected lymph nodes. Careful examination of serial sections of the SLN can however affect therapeutic decision making since staging may be upgraded in up to 10% of initially N0 patients. PMID- 17396088 TI - [Cytomegalovirus and cryptogenic inflammatory bowel disease]. AB - CMV infection has been reported in association with some flares of IBD. Its prevalence varies with the method of diagnosis and the severity of IBD. Although the link between CMV and IBD is not clear, the immunomodulator properties of the virus may play a role in the evolution of IBD. Besides the necessity of immunosuppression to treat IBD, inflammation per se can maintain in situ viral replication. Antiviral treatment can be useful in some situations. New molecular methods will permit earlier and more sensitive diagnosis of CMV infection and a better evaluation of treatment efficacy. PMID- 17396087 TI - Place of multidisciplinary consulting meetings and clinical trials in the management of colorectal cancer in France in 2000. AB - AIM: The 1998 consensus conference dealing with colon cancer, and the 2003 Cancer Plan underlined the need for multidisciplinary meetings and for including patients in therapeutic trials. The aim of this study, which pooled data from the French Cancer Registries operating within the Francim network, was to report on diagnostic and therapeutic practices in the general French population before implementation of the Cancer Plan. METHODS: The study population was composed of 2935 patients with colorectal cancer diagnosed in 2000 in twelve French administrative districts accounting for 15% of the geographical area of France. Data were collected using a standardized procedure. Three categories of place of diagnosis were defined: public university hospitals, public non-university hospitals, and private clinics. RESULTS: Overall, multidisciplinary meeting was conducted for 32.2% of patients with colorectal cancer. This proportion varied from 6.4% to 76.9%, depending on the geographical area (P<0.001). The place of diagnosis affected this practice: 52% in public university hospitals, 31% in public non-university hospitals and 29% in private clinics (P<0.001). In multivariate analysis, age (OR(>75 years): 0.71, P<0.001), site (OR(rectum): 1.80, P<0.001) and health care facilities (OR(public non-university vs public university): 0.36, P<0.001, OR(private vs public university): 0.40, P<0.001) affected the use of multidisciplinary meeting. Overall, 4.3% of patients were included in a therapeutic trial. This concerned 6.2% of patients aged under 75 and 1.0% of those aged over 75 (P<0.001). The proportion of inclusions, taking into account the trials proposed in 2000 and 2001, varied from 0.7% to 16.4% according to geographical area (P<0.001). This proportion was 10.3% if there had been multidisciplinary meeting and 5.1% if not (P<0.001). Neither cancer site, gender, nor healthcare facility responsible for diagnosis influenced trial inclusion. CONCLUSION: This population-based study underlines geographical variations in the management of colorectal cancer in France. In 2000, multidisciplinary meeting was conducted for an insufficient proportion of patients, and an insufficient number of patients were included in therapeutic trials. Repeating the same survey in 2005 will provide information on the effects of the Cancer Plan and the diffusion of these recommendations. PMID- 17396089 TI - Giant aneurysm of the main hepatic artery secondary to hereditary hemorrhagic telangiectasia: 3D contrast-enhanced MR angiography features. AB - This case report describes a patient with Hereditary Hemorrhagic Telangiectasia and a giant hepatic artery aneurysm. Aneurysms of the hepatic artery are the second most common form of visceral artery aneurysms. The causes of hepatic artery aneurysms are atherosclerosis in 30% of cases, arteritides, periarterial inflammation, liver transplantation, and hepatic tumor embolization. To our knowledge no giant hepatic artery aneurysm has been described in relation to Weber-Rendu-Osler disease in the literature. These aneurysms probably develop because of hepatic arterio-venous fistulas and secondary to changes in arterial hemodynamics. The increased use of non invasive imaging techniques such as MRI before liver transplantation in patients with hereditary hemorrhagic telangiectasia reveals these asymptomatic aneurysms and makes it possible to choose the best therapeutic approach. PMID- 17396090 TI - [Stomal varices treated by glue embolization]. AB - In patients with cirrhosis and enterostomy, although bleeding stomal varices are rare, they can been severe and difficult to be treat. We report the first two cases of bleeding stomal varices treated by cyanoacrylate embolization, in patients with cirrhosis and colostomy. In each case, after pharmacological treatment of portal hypertension (propranolol) failed, embolization of the stomal varices by transdermal injection of biological glue effectively stopped the bleeding, without recurrence or side effects, after 8 and 16 months of follow-up. The embolization of stomal varices by biological glue is a safe, easy and efficient treatment for bleeding stomal varices. PMID- 17396091 TI - [Diffuse primary fat replacement of the pancreas: an unusual cause of steatorrhea]. AB - Diffuse and primitive fat replacement of the exocrine pancreas is a rare cause of exocrine pancreatic insufficiency in adults. We report two adult patients with chronic diarrhoea and steatorrhea whose morphologic abnormalities revealed diffuse fat replacement of the pancreas. Tomodensitometry detected diffuse characteristic abnormalities with fat intermixed with normal pancreatic lobules or a "vanishing pancreas". In one patient, magnetic resonance imaging and echo endoscopy suggested abnormalities. Administration of pancreatic extracts improved symptoms. PMID- 17396092 TI - [Retroperitoneal non-secreting paraganglioma. Apropos of a case]. AB - Paragangliomas are rare tumors arising from extraadrenal chromaffin cells. These tumors are most commonly found in the adrenal gland but other locations are possible. A 79-year-old woman with abdominal pain underwent computed tomography (CT scan). Surgery was indicated because of the increase in the size of the tumor. Histopathological examination revealed a non secreting paraganglioma in the left retroperitoneum. Paragangliomas are rare neuroendocrine tumors. They have a greater potential for malignancy than pheochromocytomas due to metastases or local recurrence making surgical excision the treatment of choice. PMID- 17396093 TI - [Giant Meckel's diverticulum]. PMID- 17396094 TI - [Systemic amylosis AL revealed by a hemorrhagic gastric pseudotumor]. PMID- 17396095 TI - [Legal outcome of a case of hepatitis C nosocomial infection]. PMID- 17396096 TI - [How to treat non-responders to antiviral bitherapy for hepatitis C?]. PMID- 17396097 TI - [Treatment of hepatitis B. Current strategies]. PMID- 17396098 TI - [Can metabolic steatohepatitis be treated?]. PMID- 17396099 TI - [The role of TIPS in the treatment of refractory ascites]. PMID- 17396100 TI - [Percutaneous treatment of hepatocellular carcinoma: what is the best technique?]. PMID- 17396101 TI - [Case report: a series of autoimmune diseases]. PMID- 17396103 TI - HLA-DRB1*04 and DRB1*10 are associated with resistance and susceptibility, respectively, in Brazilian and Vietnamese leprosy patients. AB - The host genetic background has been considered one of the factors that influence leprosy outcome, a chronic infectious disease caused by Mycobacterium leprae. Genome scans demonstrated that the 6p21 region is associated with leprosy and a substantial number of population-based studies analyzing human leukocyte antigen (HLA) class II loci suggested association of HLA-DR with leprosy. However, some studies lacked robustness as they had limited power. Indeed, experimental designs require increased sample size to achieve adequate power, as well as replication studies with independent samples for confirmation of previous findings. In this work, we analyzed the influence of the HLA-DRB1 locus on leprosy susceptibility per se and disease type using a case-control design carried out in Brazilians (578 cases and 691 controls) and a replication study based on a family design in a Vietnamese population (n=194 families). The results showed that HLA-DRB1*10 is associated with susceptibility to leprosy and HLA-DRB1*04 is associated with resistance, both in the Brazilian and Vietnamese populations suggesting that these alleles play an important role in the activation of cellular immune responses against M. leprae. PMID- 17396104 TI - The lack of impact of the 'impact factor' in clinical medicine. PMID- 17396105 TI - Imatinib for the treatment of rheumatic diseases. PMID- 17396106 TI - Treatment of chronic pain in pediatric rheumatic disease. AB - Pain in children with rheumatic disease is common, and is most often caused by arthritis. Despite the widespread use of effective new biologic agents, pain continues to be a problem in these patients, and it greatly impairs their daily functioning and quality of life. The pathogenesis of pain in children with rheumatic diseases is multifactorial, and disease treatment alone is often not enough to alleviate it. No standard of care or detailed algorithm for managing pain in these patients exists. Specific pain treatments often include acetaminophen, NSAIDs and medications that treat arthritis, such as methotrexate and etanercept. Other approaches should include nonpharmacologic interventions, for example exercise and cognitive-behavioral therapy, as well as the use of analgesics such as opioids in patients whose pain is refractory to standard therapies. The use of systemic corticosteroids to treat pain in children with arthritis should be avoided. PMID- 17396107 TI - Therapy insight: scleritis and its relationship to systemic autoimmune disease. AB - The term scleritis describes a chronic inflammation that involves the outermost coat and skeleton of the eye. Disease can be isolated to the eye, but in up to half of affected individuals it occurs in the context of an immune-mediated systemic inflammatory condition, such as rheumatoid arthritis or Wegener's granulomatosis. Although uncommon, scleritis is often extremely painful, can lead to vision-threatening complications (and involvement of other ocular tissues), and is considered to confer an increased risk of mortality in patients with rheumatoid arthritis. Pathogenic mechanisms in scleritis are poorly understood, but enzymatic degradation of collagen fibrils by resident cells and infiltrating leukocytes seems to be a key feature. Several forms of inflammation can be distinguished histologically; interestingly, although the disease typically presents with engorgement of scleral vessels, vasculitis is not universally present at the microscopic level. Although some patients with scleritis respond well to treatment with NSAIDs, aggressive systemic therapy is often required to obtain a favorable outcome, particularly when systemic disease coexists. The mainstay of treatment is oral prednisone, but this agent is usually combined with a steroid-sparing immunosuppressive drug. New therapies presently under investigation for scleritis include local corticosteroid injections and various biologic agents. PMID- 17396108 TI - Drug insight: different mechanisms of action of tumor necrosis factor antagonists passive-aggressive behavior? AB - Antagonists of tumor necrosis factor (TNF) have revolutionized the treatment of selected inflammatory diseases. In rheumatology, this has been most notable for ankylosing spondylitis, psoriatic arthritis and rheumatoid arthritis. Despite their specificity for TNF, these agents, which include the soluble p75 receptor etanercept and the anti-TNF antibodies adalimumab and infliximab, have demonstrated differential clinical efficacy in studies of rheumatoid arthritis; patients who do not respond to one antagonist often respond to another. Therapeutic disparity of these agents is also seen in specific diseases, most notably Crohn's disease. Differences in pharmacodynamics, pharmacokinetics and mechanisms of action, as well as disease heterogeneity, have been proposed to account for these effects. Reverse signaling by transmembrane TNF in response to anti-TNF antibodies, but not soluble receptor, might also influence the therapeutic response. PMID- 17396109 TI - Primer: establishing a clinical trial unit - regulations and infrastructure. AB - The performance of clinical trials can be very rewarding for the practicing or academic clinical rheumatologist. There are at least 50 new compounds - small molecules and biologics - in development for rheumatic diseases such as rheumatoid arthritis, osteoarthritis, systemic lupus erythematosus, ankylosing spondylitis, psoriatic arthritis, scleroderma, gout and fibromyalgia. Clinical trials are important to try to determine the appropriate use of these compounds, as well as to answer questions about their safety. To carry out clinical trials effectively, the physician-investigator must be aware of, and adhere to, the regulatory requirements. The purpose of this article is to review these requirements in depth, as well as to discuss the infrastructure required to establish a successful clinical trial unit. PMID- 17396110 TI - Malar rash caused by metal allergy in a patient with systemic lupus erythematosus. AB - BACKGROUND: A 61-year-old woman with an 8-year history of systemic lupus erythematosus presented with a non-pruritic, erythematous, malar rash. Previously, she had tested positive for antinuclear antibody and autoantibodies to double-stranded DNA and Ro/SSA, and had an elevated erythrocyte sedimentation rate. She wore eyeglasses with metal frames and had recently gained weight, which caused the eyeglasses to have increased contact area with her face. INVESTIGATIONS: Physical examination, autoantibody testing, measurement of complement C3 and C4 levels, measurement of erythrocyte sedimentation rate, hypersensitivity patch testing, dimethylglyoxime test of the patient's eyeglass frames. DIAGNOSIS: Delayed-type hypersensitivity reaction to nickel and possibly to cobalt dichloride in the patient's eyeglass frames, which caused a malar rash that mimicked acute cutaneous lupus erythematosus. MANAGEMENT: The rash resolved completely with contact avoidance with the eyeglass frames. PMID- 17396111 TI - Renin-stimulated TGF-beta1 expression is regulated by a mitogen-activated protein kinase in mesangial cells. AB - Recent evidence indicates that renin itself might be profibrotic, independent of angiotensin II; however, the signaling system by which renin exerts a direct effect is not known. We tested the hypothesis that renin receptor activation, in turn, activates the extracellular-signal regulated kinase 1 and 2 (ERK1/2) of the mitogen-activated protein kinase system in mesangial cells. Recombinant rat renin induced a rapid phosphorylation of ERK1/2 and subsequent cell proliferation in a dose- and time-dependent manner. ERK1/2 activation by renin addition was not altered by angiotensin-converting enzyme inhibition or angiotensin receptor blockade. An ERK kinase inhibitor significantly reduced the renin-induced ERK1/2 phosphorylation and the subsequent increase in transforming growth factor-beta1 (TGF-beta1) and plasminogen activator inhibitor-1 mRNA expression. A small inhibiting RNA, siRNA, against the renin receptor completely blocked ERK1/2 activation by rat renin. We conclude that renin induces ERK1/2 activation though a receptor-mediated, angiotensin II-independent mechanism in mesangial cells. This renin-activated pathway triggers cell proliferation along with TGF-beta1 and plasminogen activator inhibitor-1 gene expression. This system may play an important role in the overall profibrotic actions of renin. PMID- 17396112 TI - Cisplatin-induced nephrotoxicity is mediated by tumor necrosis factor-alpha produced by renal parenchymal cells. AB - Cisplatin is a chemotherapeutic agent that induces tumor necrosis factor-alpha (TNF-alpha) production in many cell types with unfortunate renal toxicity. We sought to determine the contributions of renal parenchymal cells and bone marrow derived immune cells to the pathogenesis of cisplatin-induced renal injury in vivo. To do this we created chimeric mice in which the bone marrow was ablated and replaced with donor bone marrow cells from wild-type or from TNF-alpha knockout mice. Six weeks after reconstitution, the chimeric mice were treated with cisplatin and renal structural and functional parameters were measured. Chimeras with kidneys of wild-type animals all developed significant renal failure after 72 h of cisplatin treatment regardless of the immune cell source. Chimeras with kidneys of TNF-alpha knockout mice showed significantly less renal dysfunction (blood urea nitrogen, serum creatinine, and glomerular filtration rate), renal histologic injury, and serum TNF-alpha levels; again regardless of the immune cell source. Urinary excretion of several proinflammatory cytokines was lower in the wild-type bone marrow-knockout kidney chimera mouse than in wild type background mice. Our results indicate that a substantial portion of circulating and urinary TNF-alpha is derived from nonimmune cells after cisplatin administration. We conclude that the production of TNF-alpha by renal parenchymal cells, rather than by bone marrow-derived infiltrating immune cells, is responsible for cisplatin-induced nephrotoxicity. PMID- 17396113 TI - Modified RIFLE criteria in critically ill children with acute kidney injury. AB - A classification system has been proposed to standardize the definition of acute kidney injury in adults. These criteria of risk, injury, failure, loss, and end stage renal disease were given the acronym of RIFLE. We have modified the criteria based on 150 critically ill pediatric RIFLE (pRIFLE) patients to assess acute kidney injury incidence and course along with renal and/or non-renal comorbidities. Of these children, 11 required dialysis and 24 died. Patients without acute kidney injury in the first week of intensive care admission were less likely to subsequently develop renal Injury or Failure; however, 82% of acute kidney injury occurred in this initial week. Within this group of 123 children, 60 reached pRIFLEmax for Risk, 32 reached Injury, and 31 reached Failure. Acute kidney injury during admission was an independent predictor of intensive care; hospital length of stay and an increased risk of death independent of the Pediatric Risk of Mortality (PRISM II) score (odds ratio 3.0). Our results show that a majority of critically ill children develop acute kidney injury by pRIFLE criteria and do so early in the course of intensive care. Acute kidney injury is associated with mortality and may lead to increased hospital costs. We suggest that the pRIFLE criteria serves to characterize the pattern of acute kidney injury in critically ill children. PMID- 17396114 TI - Role of P-fimbrial-mediated adherence in pyelonephritis and persistence of uropathogenic Escherichia coli (UPEC) in the mammalian kidney. AB - P fimbria, a mannose-resistant adhesin of uropathogenic Escherichia coli (UPEC), has been shown to be associated with acute pyelonephritis. The pap gene cluster encodes the proteins required for P-fimbrial biogenesis, including papG, which encodes the tip adhesin. The three most studied PapG molecular variants, which are shown to bind distinct isoreceptors, are PapGI, -II, and -III. PapGII preferentially binds globoside, or GbO4, a glycolipid isoreceptor of the human kidney. Studies using different animal models of ascending urinary tract infection (UTI) have demonstrated a variable role for P fimbriae, and specifically PapGII-mediated adherence, in renal colonization. The disparities in the results obtained from those studies are likely to be attributed to the differences in animal models and UPEC strains utilized. One explanation that is discussed in detail is the contribution of multiple fimbriae of UPEC that potentially mediate adherence to the mammalian kidney. Overall, P fimbriae appear to play some role in mediating adherence to uroepithelial cells in vivo and establishing an inflammatory response during renal colonization, thus contributing to kidney damage during acute pyelonephritis. To verify that P fimbriae contribute to the pathogenesis of UPEC during ascending UTI (and in particular acute pyelonephritis), future studies should be conducted to satisfy fully all three tenets of the molecular Koch's postulates, including complementation of a mutated allele. PMID- 17396115 TI - Hyperproliferation of PKD1 cystic cells is induced by insulin-like growth factor 1 activation of the Ras/Raf signalling system. AB - Autosomal dominant polycystic kidney disease (ADPKD) largely results from mutations in the PKD1 gene leading to hyperproliferation of renal tubular epithelial cells and consequent cyst formation. Rodent models of PKD suggest that the multifunctional hormone insulin-like growth factor-1 (IGF-1) could play a pathogenic role in renal cyst formation. In order to test this possibility, conditionally immortalized renal epithelial cells were prepared from normal individuals and from ADPKD patients with known germline mutations in PKD1. All patient cell lines had a decreased or absence of polycystin-1 but not polycystin 2. These cells had an increased sensitivity to IGF-1 and to cyclic AMP, which required phosphatidylinositol-3 (PI3)-kinase and the mitogen-activated protein kinase, extracellular signal-regulated protein kinase (ERK) for enhanced growth. Inhibition of Ras or Raf abolished the stimulated cell proliferation. Our results suggest that haploinsufficiency of polycystin-1 lowers the activation threshold of the Ras/Raf signalling system leading to growth factor-induced hyperproliferation. Inhibition of Ras or Raf activity may be a therapeutic option for decreasing tubular cell proliferation in ADPKD. PMID- 17396116 TI - Conversion from conventional to nocturnal hemodialysis improves vitamin D levels. AB - Patients on conventional hemodialysis have low levels of 25-hydroxy-vitamin D probably due to diet and decreased cutaneous synthesis. As 1,25 dihydroxy-vitamin D synthesis is substrate-dependent in end-stage renal disease, this could be a contributing factor to low 1,25 dihydroxy-vitamin D levels in patients undergoing conventional hemodialysis. We converted 35 patients historically on conventional hemodialysis to nocturnal hemodialysis for a minimum of 6 months thereby significantly increasing sessional equilibrated Kt/V from an average of 1.30 to an average of 2.01. Dietary restrictions were also removed. Serum phosphorus significantly fell, whereas the serum calcium, parathyroid hormone, and the mean dose of calcitriol did not change after the conversion. Significant increases in both 25-hydroxy and 1,25-dihydroxy-vitamin D levels were seen after hemodialysis mode conversion. A significant correlation was found between the dialysis dose and the levels of both hydroxylated forms of vitamin D. We suggest that improving uremia by nocturnal hemodialysis in the absence of exogenous supplementation is associated with increased 25 and 1,25-hydroxy-vitamin D levels. Additionally, normalization of serum phosphorus may improve 1alpha-hydroxylation thereby enhancing substrate-dependent generation of 1,25-dihydroxy-vitamin D in chronic dialysis patients. PMID- 17396117 TI - Acute kidney injury: changing lexicography, definitions, and epidemiology. AB - In recent years, there have been numerous advances in understanding the molecular determinants of functional kidney injury after ischemic and/or toxic exposure. However, translation of successful novel therapies designed to attenuate kidney functional injury from animal models to the clinical sphere has had modest results. This lack of translatability is at least in part due to lack of sufficient standardization in definitions and classification of cases of acute kidney injury (AKI), an incomplete understanding of the natural history of human AKI, and a limited understanding of how kidney injury interacts with other organ system failure in the context of systemic metabolic abnormalities. A concerted effort is now being made by nephrologists and intensivists to arrive at standardized terminology and classification of AKI. There have also been dramatic advances in our understanding of the epidemiology and natural history of AKI, particularly in the hospital and intensive care unit setting. Promising strategies are now being developed which may ultimately lead to improved outcomes for patients at risk for or who have developed AKI, which should be readily testable in the coming decade. PMID- 17396118 TI - Predictors of the response to treatment in anemic hemodialysis patients with high serum ferritin and low transferrin saturation. AB - Treating hemodialysis patients to combat anemia corrects hemoglobin but exacerbates iron deficiency by utilizing iron stores. Patients needing iron should receive this by intravenous (i.v.) means. The Dialysis patients' Response to IV iron with Elevated ferritin (DRIVE) trial investigated the role of i.v. iron in anemic patients with high ferritin, low transferrin saturation, and adequate epoetin doses. We examined whether baseline iron and inflammation markers predict the response of hemoglobin to treatment. Patients (134) were randomized to no added iron or to i.v. ferric gluconate for eight consecutive hemodialysis sessions spanning 6 weeks with epoetin increased by 25% in both groups. The patients started with hemoglobin less than or equal to 11 g/dl, ferritin between 500 and 1200 ng/ml, and transferrin saturation of less than 25%. Significantly, patients with a reticulocyte hemoglobin content greater than or equal to 31.2 pg were over five times more likely to achieve a clinically significant increase in hemoglobin of greater than 2 g/dl. Lower reticulocyte hemoglobin contents did not preclude a response to i.v. iron. Significantly higher transferrin saturation or lower C-reactive protein but not ferritin or soluble transferrin receptor levels predicted a greater response; however their influence was not clinically significant in either group. We conclude that none of the studied markers is a good predictor of response to anemia treatment in this patient sub-population. PMID- 17396119 TI - Sixteen novel mutations identified in COL4A3, COL4A4, and COL4A5 genes in Slovenian families with Alport syndrome and benign familial hematuria. AB - Alport syndrome (ATS) and benign familial hematuria (BFH) are type IV collagen inherited disorders. Mutations in COL4A5 are generally believed to cause X-linked ATS, whereas mutations in COL4A3 and COL4A4 genes can be associated with the autosomal-recessive and -dominant type of ATS or BFH. In view of the wide spectrum of phenotypes, an exact diagnosis is sometimes difficult to achieve. This study involved screening each exon with boundary intronic sequences of COL4A3, COL4A4, and COL4A5 genes by optimized polymerase chain reaction-single stranded conformational polymorphism analysis in 17 families with ATS and in 40 families diagnosed as having BFH. Twelve different mutations were found in the COL4A5 gene in ATS patients, comprising nine missense mutations, a splice site mutation, a mutation causing frameshift, and a nonsense mutation. One of the missense mutations (p.G624D) was present not only in one family with ATS but also in five families with suspected BFH. Three heterozygous mutations in the COL4A3 gene (two missense and one frameshift) and four heterozygous mutations in COL4A4 (two splice site, one in-frame deletion, and one missense) were identified in patients with BFH. Sixteen mutations are to the best of our knowledge new and private. PMID- 17396120 TI - Raf kinase signaling functions in sensory neuron differentiation and axon growth in vivo. AB - To define the role of the Raf serine/threonine kinases in nervous system development, we conditionally targeted B-Raf and C-Raf, two of the three known mammalian Raf homologs, using a mouse line expressing Cre recombinase driven by a nestin promoter. Targeting of B-Raf, but not C-Raf, markedly attenuated baseline phosphorylation of Erk in neural tissues and led to growth retardation. Conditional elimination of B-Raf in dorsal root ganglion (DRG) neurons did not interfere with survival, but instead caused marked reduction in expression of the glial cell line-derived neurotrophic factor receptor Ret at postnatal stages, associated with a profound reduction in levels of transcription factor CBF-beta. Elimination of both alleles of Braf, which encodes B-Raf, and one allele of Raf1, which encodes C-Raf, affected DRG neuron maturation as well as proprioceptive axon projection toward the ventral horn in the spinal cord. Finally, conditional elimination of all Braf and Raf1 alleles strongly reduced neurotrophin-dependent axon growth in vitro as well as cutaneous axon terminal arborization in vivo. We conclude that Raf function is crucial for several aspects of DRG neuron development, including differentiation and axon growth. PMID- 17396121 TI - Memory consolidation or transformation: context manipulation and hippocampal representations of memory. AB - The traditional view is that the hippocampus is necessary for retaining memories until they are consolidated in their original form in the neocortex. An alternative view is that the original memory, which is hippocampus- and context dependent, becomes transformed with time to one that is more schematic and independent of the hippocampus. By manipulating context in two protocols that are widely used to investigate hippocampal-neocortical interactions in memory, we find evidence for the transformation view. PMID- 17396122 TI - Human visual cortex responds to invisible chromatic flicker. AB - When two isoluminant colors alternate at frequencies of 25 Hz or higher, observers perceive only one fused color. Chromatic flicker beyond the fusion frequency induces flicker adaptation in human observers and stimulates monkey V1 neurons. Here we use functional magnetic resonance imaging (fMRI) to show that many human visual cortical areas, with the exception of VO, can distinguish between fused chromatic flicker and its matched nonflickering control. This result supports the existence of significant intracortical temporal filtering of high-frequency chromatic information. The result also suggests that a considerable difference in cortical activation in many visual cortical areas does not necessarily lead to different conscious experiences. PMID- 17396123 TI - The proprioceptive representation of eye position in monkey primary somatosensory cortex. AB - The cerebral cortex must have access to an eye position signal, as humans can report passive changes in eye position in total darkness, and visual responses in many cortical areas are modulated by eye position. The source of this signal is unknown. Here we demonstrate a representation of eye position in monkey primary somatosensory cortex, in the representation of the trigeminal nerve, near cells with a tactile representation of the contralateral brow. The neurons have eye position signals that increase monotonically with increasing orbital eccentricity from near the center of gaze, with directionally selectivity tuned in a Gaussian manner. All directions of eye position are represented in a single hemisphere. The signal is proprioceptive, because it can be obliterated by anesthetizing the contralateral orbit. It is not related to foveal or peripheral visual stimulation, and it represents the position of the eye in the head and not the angle of gaze in space. PMID- 17396124 TI - TRPC channels promote cerebellar granule neuron survival. AB - Channels formed by the transient receptor potential (TRP) family of proteins have a variety of physiological functions. Here we report that two members of the TRP cation channel (TRPC) subfamily, TRPC3 and 6, protected cerebellar granule neurons (CGNs) against serum deprivation-induced cell death in cultures and promoted CGN survival in rat brain. In CGN cultures, blocking TRPC channels or downregulating TRPC3 or 6 suppressed brain-derived neurotrophic factor (BDNF) mediated protection, BDNF-triggered intracellular Ca2+ elevation and BDNF-induced CREB activation. By contrast, overexpressing TRPC3 or 6 increased CREB-dependent reporter gene transcription and prevented apoptosis in the neurons deprived of serum, and this protection was blocked by the dominant negative form of CREB. Furthermore, downregulating TRPC3 or 6 induced CGN apoptosis in neonatal rat cerebellum, and this effect was rescued by overexpressing either TRPC3 or 6. Thus, our findings provide in vitro and in vivo evidence that TRPC channels are important in promoting neuronal survival. PMID- 17396125 TI - The rise of the 'projectome'. PMID- 17396126 TI - Do-it-yourself RNAi made easy? PMID- 17396127 TI - Three-dimensional culture models of normal and malignant breast epithelial cells. AB - Extracellular matrix is a key regulator of normal homeostasis and tissue phenotype. Important signals are lost when cells are cultured ex vivo on two dimensional plastic substrata. Many of these crucial microenvironmental cues may be restored using three-dimensional (3D) cultures of laminin-rich extracellular matrix (lrECM). These 3D culture assays allow phenotypic discrimination between nonmalignant and malignant mammary cells, as the former grown in a 3D context form polarized, growth-arrested acinus-like colonies whereas the latter form disorganized, proliferative and nonpolar colonies. Signaling pathways that function in parallel in cells cultured on plastic become reciprocally integrated when the cells are exposed to basement membrane-like gels. Appropriate 3D culture thus provides a more physiologically relevant approach to the analysis of gene function and cell phenotype ex vivo. We describe here a robust and generalized method for the culturing of various human breast cell lines in three dimensions and describe the preparation of cellular extracts from these cultures for molecular analyses. The procedure below describes the 3D 'embedded' assay, in which cells are cultured embedded in an lrECM gel (Fig. 1). By lrECM, we refer to the solubilized extract derived from the Engelbreth-Holm-Swarm mouse sarcoma cells. For a discussion of user options regarding 3D matrices, see Box 1. Alternatively, the 3D 'on-top' assay, in which cells are cultured on top of a thin lrECM gel overlaid with a dilute solution of lrECM, may be used as described in Box 2 (Fig. 1 and Fig. 2). PMID- 17396129 TI - Reversal of renal disease: is it enough to inhibit the action of angiotensin II? AB - Over the last years, evidence emerged demonstrating that the progression of renal fibrosis is reversible in experimental models. The present review summarizes the new insights concerning the mechanisms of progression and regression of renal disease and examines this novel evidence under the light of feasibility and transfer to human nephropathies. The involved mechanisms are discussed with particular emphasis on the fibrotic role of vasoactive peptides such as angiotensin II and endothelin, and growth factors such as transforming growth factor beta (TGFbeta). The possibility of regression is introduced by presenting the in vivo efficiency of anti-hypertensive treatments and of systems that antagonize the fibrogenic action of TGFbeta such as bone morphogenic protein-7 (BMP-7) and hepatocyte growth factor. Finally, we provide a brief description of the promising future directions and clinical considerations about the applications of the experimental data to humans. PMID- 17396130 TI - Migration, fate and in vivo imaging of adult stem cells in the CNS. AB - Adult stem cells have been intensively studied for their potential use in cell therapies for neurodegenerative diseases, ischemia and traumatic injuries. One of the most promising cell sources for autologous cell transplantation is bone marrow, containing a heterogenous cell population that can be roughly divided into hematopoietic stem and progenitor cells and mesenchymal stem cells (MSCs). MSCs are multipotent progenitor cells that, in the case of severe tissue ischemia or damage, can be attracted to the lesion site, where they can secrete bioactive molecules, either naturally or through genetic engineering. They can also serve as vehicles for delivering therapeutic agents. Mobilized from the marrow, sorted or expanded in culture, MSCs can be delivered to the damaged site by direct or systemic application. In addition, MSCs can be labeled with superparamagnetic nanoparticles that allow in vivo cell imaging. Magnetic resonance imaging (MRI) is thus a suitable method for in vivo cell tracking of transplanted cells in the host organism. This review will focus on cell labeling for MRI and the use of MSCs in experimental and clinical studies for the treatment of brain and spinal cord injuries. PMID- 17396131 TI - Adenosine, an endogenous distress signal, modulates tissue damage and repair. AB - Adenosine is formed inside cells or on their surface, mostly by breakdown of adenine nucleotides. The formation of adenosine increases in different conditions of stress and distress. Adenosine acts on four G-protein coupled receptors: two of them, A(1) and A(3), are primarily coupled to G(i) family G proteins; and two of them, A(2A) and A(2B), are mostly coupled to G(s) like G proteins. These receptors are antagonized by xanthines including caffeine. Via these receptors it affects many cells and organs, usually having a cytoprotective function. Joel Linden recently grouped these protective effects into four general modes of action: increased oxygen supply/demand ratio, preconditioning, anti-inflammatory effects and stimulation of angiogenesis. This review will briefly summarize what is known and what is not in this regard. It is argued that drugs targeting adenosine receptors might be useful adjuncts in many therapeutic approaches. PMID- 17396132 TI - Endoplasmic reticulum stress-induced cell death mediated by the proteasome. AB - Cells exposed to sustained endoplasmic reticulum (ER) stress undergo programmed cell death and display features typical of apoptosis, such as cysteine aspartyl protease (caspase) activation, cytochrome c release, and DNA fragmentation. Here, we show that the execution of cell death induced by ER stress is mediated via the proteasome. Inhibition of the proteasome by lactacystin prevented ER stress induced degradation of Bcl-2, release of cytochrome c, processing of effector caspase-3, and exposure of phosphatidylserine. Owing to the ability of lactacystin to inhibit cytochrome c release, we propose that the pro-apoptotic activity of the proteasome lies upstream of mitochondrial activation. Thus, the proteasome serves as a principal mediator of ER stress-induced cell death in this system. PMID- 17396134 TI - Angiogenesis: an organizing principle for drug discovery? AB - Angiogenesis--the process of new blood-vessel growth--has an essential role in development, reproduction and repair. However, pathological angiogenesis occurs not only in tumour formation, but also in a range of non-neoplastic diseases that could be classed together as 'angiogenesis-dependent diseases'. By viewing the process of angiogenesis as an 'organizing principle' in biology, intriguing insights into the molecular mechanisms of seemingly unrelated phenomena might be gained. This has important consequences for the clinical use of angiogenesis inhibitors and for drug discovery, not only for optimizing the treatment of cancer, but possibly also for developing therapeutic approaches for various diseases that are otherwise unrelated to each other. PMID- 17396135 TI - Potential therapeutic applications of autophagy. AB - Autophagy is a dynamic process of subcellular degradation, which has recently sparked great interest as it is now recognized to be involved in various developmental processes and various diseases including cancer and neurodegeneration. Autophagy can function as a cytoprotective mechanism; however, it also has the capacity to cause cell death. A better understanding of autophagy is needed to allow its manipulation for therapeutic purposes, and new insights into the molecular mechanisms of autophagy are now leading to the discovery of exciting new potential drug targets. PMID- 17396136 TI - Antagonism of the prostaglandin D2 receptors DP1 and CRTH2 as an approach to treat allergic diseases. AB - Immunological activation of mast cells is an important trigger in the cascade of inflammatory events leading to the manifestation of allergic diseases. Pharmacological studies using the recently discovered DP(1) and CRTH2 antagonists combined with genetic analysis support the view that these receptors have a pivotal role in mediating aspects of allergic diseases that are resistant to current therapy. This Review focuses on the emerging roles that DP(1) and CRTH2 (also known as DP(2)) have in acute and chronic aspects of allergic diseases and proposes that, rather than having opposing actions, these receptors have complementary roles in the initiation and maintenance of the allergy state. We also discuss recent progress in the discovery and development of selective antagonists of these receptors. PMID- 17396137 TI - Telomere dysfunction suppresses spontaneous tumorigenesis in vivo by initiating p53-dependent cellular senescence. AB - Dysfunctional telomeres induce p53-dependent cellular senescence and apoptosis, but it is not known which function is more important for tumour suppression in vivo. We used the p53 ( R172P ) knock-in mouse, which is unable to induce apoptosis but retains intact cell-cycle arrest and cellular senescence pathways, to show that spontaneous tumorigenesis is potently repressed in Terc -/- p53 ( R172P ) mice. Tumour suppression is accompanied by global induction of p53, p21 and the senescence marker senescence-associated-beta-galactosidase. By contrast, cellular senescence was unable to suppress chemically induced skin carcinomas. These results indicate that suppression of spontaneous tumorigenesis by dysfunctional telomeres requires the activation of the p53-dependent cellular senescence pathway. PMID- 17396138 TI - S100A1 and S100B, transcriptional targets of SOX trio, inhibit terminal differentiation of chondrocytes. AB - Transcription factor SOX9 (sex-determining region Y-type high mobility group box 9) and its coactivators SOX5 and SOX6 (the SOX trio) induce early-stage chondrocyte differentiation and suppress its terminal stage. To identify possible targets of the SOX trio, we carried out a microarray analysis and identified S100A1 and S100B as possible target molecules. S100 protein expression was localized in late proliferative and pre-hypertrophic chondrocytes of the mouse growth plate. Overexpression of S100A1, S100B or their combination in cultured chondrogenic cells did not induce early differentiation, but suppressed hypertrophic differentiation and mineralization. Silencing of both S100A1 and S100B stimulated terminal differentiation and reversed the SOX-trio-mediated inhibition. Finally, luciferase reporter, electrophoretic mobility shift and chromatin immunoprecipitation analyses showed that transcription of both S100 proteins is induced by the SOX trio, and also identified their respective enhancer elements in the 5'-end flanking region. We conclude that S100A1 and S100B are transcriptional targets of the SOX trio and mediate its inhibition of terminal differentiation of chondrocytes. PMID- 17396139 TI - An in vitro cytologic assay for evaluation of the KIT signaling pathway in gastrointestinal stromal tumors. AB - Specific inhibitors can be designed to inactivate the molecular pathways involved in tumor growth. A compelling example is the use of small molecule drugs, such as imatinib (Gleevec), which inhibit the KIT tyrosine kinase in gastrointestinal stromal tumors (GIST). Assays are needed to determine which inhibitor is most effective at silencing the KIT kinase in each GIST patient. The aim of this study was to develop a robust, cytology-based assay to measure tumor susceptibility to target-specific small molecule inhibitors. We created an immortal GIST cell line (GIST882) that was treated in vitro with several inhibitors of the KIT --> AKT - > mTOR --> S6 signaling pathway. KIT was inhibited with imatinib, and mTOR with RAD001. Treatment response was assessed in cytologic preparations by immunocytochemical staining with antibodies to KIT, phospho-KIT, phospho-AKT, and phospho-S6. Optimization was performed to maximize staining in the absence of inhibitor, and minimize staining in the presence of inhibitor. GIST882 cells demonstrated strong, robust phospho-S6 expression in the absence of inhibitor. This expression was completely inhibited by treatment with upstream signaling pathway inhibitors (imatinib and RAD001). Other phospho-specific antibodies had weaker baseline reactivity in the absence of inhibitor. The accuracy of the immunocytochemical results on the cytologic preparations was validated by immunoblotting studies. Our study demonstrates the feasibility of cytologic methods to monitor labile biochemical responses in tumor cells during drug therapy. Such approaches will be enhanced by the development of additional activation state-specific antibodies, particularly those optimized for use in cytologic preparations. PMID- 17396140 TI - Utility of ALK-1 protein expression and ALK rearrangements in distinguishing inflammatory myofibroblastic tumor from malignant spindle cell lesions of the urinary bladder. AB - Inflammatory myofibroblastic tumor of the urinary bladder is an unusual spindle cell neoplasm that displays cytologic atypia, infiltrative growth and mitotic activity mimicking malignant tumors, such as leiomyosarcoma, rhabdomyosarcoma and sarcomatoid carcinoma. The objective of this study was to determine if anaplastic lymphoma kinase (ALK-1) protein expression detected by immunohistochemistry and ALK rearrangements detected by fluorescence in situ hybridization (FISH) were useful in distinguishing inflammatory myofibroblastic tumor from malignant spindle cell tumors of the urinary bladder. In inflammatory myofibroblastic tumor, ALK-1 expression was identified in 13 of 21 cases (62%) and ALK rearrangements in 14 of 21 cases (67%). All cases of inflammatory myofibroblastic tumor demonstrating ALK-1 expression, carried ALK rearrangements. One case negative for ALK-1 expression exhibited ALK rearrangement. ALK rearrangements were more common in women (P=0.0032). Leiomyosarcoma, sarcomatoid carcinoma, embryonal rhabdomyosarcoma and reactive myofibroblastic proliferations were negative for ALK-1 protein and ALK rearrangements. Immunohistochemistry using markers of muscle, epithelial, neural, and follicular dendritic cell differentiation showed overlap between inflammatory myofibroblastic tumor with and without ALK gene rearrangements, and between inflammatory myofibroblastic tumor and spindle cell malignancies. However, coexpression of cytokeratin and muscle-specific antigens was unique to inflammatory myofibroblastic tumor, observed in approximately half the tumors. This study indicates that detection of ALK protein and ALK gene rearrangements are useful in distinguishing inflammatory myofibroblastic tumor from spindle cell malignancies in the urinary bladder. Additionally, our findings suggest that ALK rearrangement is the primary mechanism for ALK activation and that inflammatory myofibroblastic tumor likely represents a heterogeneous group of spindle cell proliferations with the majority associated with ALK translocations, and the remaining associated with other etiologies. PMID- 17396141 TI - Standardization of HER2 testing: results of an international proficiency-testing ring study. AB - Human epidermal growth factor receptor 2 (HER2) positivity in breast cancer is a prognostic factor regarding tumor aggressiveness and a predictive factor for response to trastuzumab (Herceptin). Early and accurate HER2 testing of all breast cancer patients at primary diagnosis is essential for optimal disease management. Routine HER2 tests, such as immunohistochemistry and fluorescence in situ hybridization (FISH), are subject to interlaboratory variation, and validation by laboratory proficiency testing is important to improve standardization. This study compared immunohistochemistry and FISH testing between five international pathology reference centers. Each center evaluated 20 immunohistochemistry and 20 FISH breast cancer specimens in five testing rounds. In each round, one center selected two sets of four different invasive tumor specimens (set A for immunohistochemistry and set B for FISH) and sent samples to the other four centers in a blinded manner, while retaining samples for its own evaluation. Results were analyzed by an independent coordinator. With immunohistochemistry, there were no differences between the five centers for any of the specimens at the level of diagnostic decision (positive or negative HER2 status). However, differences between laboratories were observed in immunohistochemistry scoring. Of the 20 specimens, four were scored as negative (0/1+) and five as positive (3+) in all centers; eight were negative or equivocal (2+), and three positive or equivocal. After FISH retesting of nine of the 11 equivocal immunohistochemistry cases, consensus was achieved in 15 of 18 (83%) specimens. FISH analysis of set B specimens resulted in consensus between centers in 16 of 20 (80%) specimens (six negative and 10 positive). All four discordant FISH specimens were scored as having HER2:CEP17 ratios within the range 1.7-2.3 by at least one center. Equivocal immunohistochemistry and borderline FISH cases are difficult to interpret, even for highly experienced and validated laboratories, highlighting the need for quality-control procedures. PMID- 17396142 TI - Dense deposit disease is not a membranoproliferative glomerulonephritis. AB - Dense deposit disease (first reported in 1962) was classified as subtype II of membranoproliferative glomerulonephritis in the early 1970s. Over the last 30 years, marked differences in etiology and pathogenesis between type I membranoproliferative glomerulonephritis and dense deposit disease have become apparent. The sporadic observation that dense deposit disease can be seen with markedly different light microscopy appearances prompted this study. The goal was to examine a large number of renal biopsies from around the world to characterize the histopathologic features of dense deposit disease. Eighty-one cases of dense deposit disease were received from centers across North America, Europe and Japan. Biopsy reports, light microscopy materials and electron photomicrographs were reviewed and histopathologic features scored. Sixty-nine cases were acceptable for review. Five patterns were seen: (1) membranoproliferative n=17; (2) mesangial proliferative n=30; (3) crescentic n=12; (4) acute proliferative and exudative n=8 and (5) unclassified n=2. The age range was 3-67 years, with 74% in the range of 3-20 years; 15% 21-30 years and 11% over 30 years. Males accounted for 54% and females 46%. All patients with either crescentic dense deposit disease or acute proliferative dense deposit disease were between the ages of 3 and 18 years. The essential diagnostic feature of dense deposit disease is not the membranoproliferative pattern but the presence of electron dense transformation of the glomerular basement membranes. Based upon this study and the extensive data developed over the past 30 years, dense deposit disease is clinically distinct from membranoproliferative glomerulonephritis and is morphologically heterogeneous with only a minority of cases having a membranoproliferative pattern. Therefore, dense deposit disease should no longer be regarded as a subtype of membranoproliferative glomerulonephritis. PMID- 17396143 TI - Prognostic significance of maspin in pancreatic ductal adenocarcinoma: tissue microarray analysis of 223 surgically resected cases. AB - Maspin (SERPINB5), a serine proteinase inhibitor, was first identified as a potential tumor suppressor on the basis of its differential expression between normal mammary epithelial cells and human breast carcinoma cell lines. Recent studies have shown that maspin might be a prognostic tumor marker. Pancreatic ductal adenocarcinoma acquires maspin expression through hypomethylation of the maspin promoter. However, no study has investigated the prognostic significance of maspin expression in pancreatic ductal adenocarcinomas. In this study, we investigated maspin protein expression in a large series of 223 surgically resected pancreatic ductal adenocarcinomas using immunohistochemical staining and high throughput tissue microarrays. Maspin expression was correlated with postoperative survival and other clinicopathologic factors. Maspin was detected in 209 of these 223 (94% cases) pancreatic ductal adenocarcinomas including 39 (18% cases) focal (5-50% tumor cells) and 170 (76% cases) diffuse (>50% tumor cells). Fourteen (or 6% cases) pancreatic ductal adenocarcinomas did not show maspin expression by immunohistochemical staining (<5% tumor cells). Normal ductal epithelium is not labeled with maspin. Overexpression of maspin in pancreatic ductal adenocarcinoma is associated with worse postoperative survival especially in patients whose tumors exhibit diffuse expression of maspin. After adjusting other clinicopathologic factors, maspin expression remains to be an independent adverse prognosticator for postoperative survival. Maspin expression is not associated with patient age, gender, tumor size, tumor pathologic stage, lymph node status, and vascular invasion or perineural invasion. Nuclear labeling of maspin is associated with better tumor differentiation although this staining pattern is not associated with a better prognosis. In addition, maspin overexpression is also observed in 48% low-grade (grades 1a and 1b) pancreatic intraepithelial neoplasias (PanINs) and 78% high-grade (grades 2 and 3) PanINs, suggesting that maspin upregulation occurs early during the multi-step progression model of pancreatic ductal adenocarcinoma. PMID- 17396144 TI - Clinical and prognostic significances of nuclear and cytoplasmic KIT expressions in extrahepatic bile duct carcinomas. AB - After receiving FDA approval as a therapeutic regimen in gastrointestinal stromal tumors, the tyrosine kinase inhibitor imatinib mesylate has been applied to the treatment of other solid malignant neoplasms. To evaluate the usefulness of imatinib mesylate as a possible therapeutic regimen in extrahepatic bile duct carcinomas, an immunohistochemical study for KIT was performed in 289 cases of extrahepatic bile duct carcinomas, and mutational analysis of exon 11 of the c kit gene was performed in 20 cases that were arbitrarily retrieved from the cases with KIT expression. Cytoplasmic KIT expression was observed in 54 cases (19%) and nuclear KIT in 58 cases (20%) of extrahepatic bile duct carcinoma. Nuclear KIT expression was more frequent in cases with vascular invasion (P<0.001), whereas cytoplasmic KIT expression was more common in tumors of T1-T3 than in those of T4 (P=0.04), and was more frequently observed in cases with a papillary growth pattern (P=0.03). Patients with cytoplasmic KIT-positive tumors had significantly better survival both by univariate (P=0.01) and multivariate analyses (P=0.04). Infrequent cytoplasmic KIT expression without mutation of exon 11 suggests that imatinib mesylate may not be effective for the treatment of extrahepatic bile duct carcinoma. However, immunohistochemical study for KIT may be helpful in routine pathologic examinations for evaluating better prognosis for patients with extrahepatic bile duct carcinoma. In addition, more frequent nuclear expression of KIT in cases with vascular invasion suggests that nuclear KIT expression may contribute to the progression of extrahepatic bile duct carcinoma. PMID- 17396145 TI - Fascin overexpression in intraductal papillary mucinous neoplasms (adenomas, borderline neoplasms, and carcinomas) of the pancreas, correlated with increased histological grade. AB - Intraductal papillary mucinous neoplasm (IPMN) is a well-established entity in pancreatic neoplasms and a precursor of infiltrating adenocarcinoma. Fascin, an actin-bundling protein involved in cellular motility, is upregulated in many human neoplasms. Its overexpression in pancreatic intraepithelial neoplasia, a pre-cancerous lesion sharing many characteristics with IPMN, has been reported. However, fascin expression in IPMN remains unknown. The aim of this study was to investigate fascin expression in IPMNs and to elucidate its relationship to clinicopathological features, including histological grade and phenotypic subclassification. We evaluated fascin expression by immunohistochemistry in 116 surgical specimens, followed by quantitative analysis of fascin mRNA expression using a laser microdissection system and real-time reverse-transcriptase polymerase chain reaction in eight frozen samples. Fascin expression was significantly higher in borderline neoplasms (25/29, 86%) and carcinomas (37/42, 88%) than in adenomas (23/45, 51%) (P<0.05, respectively), but no difference was observed between borderline neoplasms and carcinomas. With regard to the subclassification, intestinal-type neoplasms (35/39, 90%) were more frequently positive for fascin than gastric-type neoplasms (36/59, 61%) (P<0.05). Two oncocytic-type neoplasms were both fascin-negative. Fascin mRNA expression seemed to be higher in moderately to severely dysplastic epithelium than in mildly dysplastic epithelium (not statistically significant), supporting the immunohistochemical experiments. Our findings suggest that fascin overexpression is involved in the progression of IPMN. Fascin could become a new therapeutic target for inhibition of their progression. PMID- 17396146 TI - TAO kinases mediate activation of p38 in response to DNA damage. AB - Thousand and one amino acid (TAO) kinases are Ste20p-related MAP kinase kinase kinases (MAP3Ks) that activate p38 MAPK. Here we show that the TAO kinases mediate the activation of p38 in response to various genotoxic stimuli. TAO kinases are activated acutely by ionizing radiation, ultraviolet radiation, and hydroxyurea. Full-length and truncated fragments of dominant negative TAOs inhibit the activation of p38 by DNA damage. Inhibition of TAO expression by siRNA also decreases p38 activation by these agents. Cells in which TAO kinases have been knocked down are less capable of engaging the DNA damage-induced G2/M checkpoint and display increased sensitivity to IR. The DNA damage kinase ataxia telangiectasia mutated (ATM) phosphorylates TAOs in vitro; radiation induces phosphorylation of TAO on a consensus site for phosphorylation by the ATM protein kinase in cells; and TAO and p38 activation is compromised in cells from a patient with ataxia telangiectasia that lack ATM. These findings indicate that TAO kinases are regulators of p38-mediated responses to DNA damage and are intermediates in the activation of p38 by ATM. PMID- 17396147 TI - FAAP100 is essential for activation of the Fanconi anemia-associated DNA damage response pathway. AB - The Fanconi anemia (FA) core complex plays a central role in the DNA damage response network involving breast cancer susceptibility gene products, BRCA1 and BRCA2. The complex consists of eight FA proteins, including a ubiquitin ligase (FANCL) and a DNA translocase (FANCM), and is essential for monoubiquitination of FANCD2 in response to DNA damage. Here, we report a novel component of this complex, termed FAAP100, which is essential for the stability of the core complex and directly interacts with FANCB and FANCL to form a stable subcomplex. Formation of this subcomplex protects each component from proteolytic degradation and also allows their coregulation by FANCA and FANCM during nuclear localization. Using siRNA depletion and gene knockout techniques, we show that FAAP100-deficient cells display hallmark features of FA cells, including defective FANCD2 monoubiquitination, hypersensitivity to DNA crosslinking agents, and genomic instability. Our study identifies FAAP100 as a new critical component of the FA-BRCA DNA damage response network. PMID- 17396148 TI - Genomic instability of the host cell induced by the human papillomavirus replication machinery. AB - Development of invasive cervical cancer upon infection by 'high-risk' human papillomavirus (HPV) in humans is a stepwise process in which some of the initially episomal 'high-risk' type of HPVs (HR-HPVs) integrate randomly into the host cell genome. We show that HPV replication proteins E1 and E2 are capable of inducing overamplification of the genomic locus where HPV origin has been integrated. Clonal analysis of the cells in which the replication from integrated HPV origin was induced showed excision, rearrangement and de novo integration of the HPV containing and flanking cellular sequences. These data suggest that papillomavirus replication machinery is capable of inducing genomic changes of the host cell that may facilitate the formation of the HPV-dependent cancer cell. PMID- 17396149 TI - Electron cryotomography of immature HIV-1 virions reveals the structure of the CA and SP1 Gag shells. AB - The major structural elements of retroviruses are contained in a single polyprotein, Gag, which in human immunodeficiency virus type 1 (HIV-1) comprises the MA, CA, spacer peptide 1 (SP1), NC, SP2, and p6 polypeptides. In the immature HIV-1 virion, the domains of Gag are arranged radially with the N-terminal MA domain at the membrane and C-terminal NC-SP2-p6 region nearest to the center. Here, we report the three-dimensional structures of individual immature HIV-1 virions, as obtained by electron cryotomography. The concentric shells of the Gag polyprotein are clearly visible, and radial projections of the different Gag layers reveal patches of hexagonal order within the CA and SP1 shells. Averaging well-ordered unit cells leads to a model in which each CA hexamer is stabilized by a bundle of six SP1 helices. This model suggests why the SP1 spacer is essential for assembly of the Gag lattice and how cleavage between SP1 and CA acts as a structural switch controlling maturation. PMID- 17396150 TI - A novel human AP endonuclease with conserved zinc-finger-like motifs involved in DNA strand break responses. AB - DNA damage causes genome instability and cell death, but many of the cellular responses to DNA damage still remain elusive. We here report a human protein, PALF (PNK and APTX-like FHA protein), with an FHA (forkhead-associated) domain and novel zinc-finger-like CYR (cysteine-tyrosine-arginine) motifs that are involved in responses to DNA damage. We found that the CYR motif is widely distributed among DNA repair proteins of higher eukaryotes, and that PALF, as well as a Drosophila protein with tandem CYR motifs, has endo- and exonuclease activities against abasic site and other types of base damage. PALF accumulates rapidly at single-strand breaks in a poly(ADP-ribose) polymerase 1 (PARP1) dependent manner in human cells. Indeed, PALF interacts directly with PARP1 and is required for its activation and for cellular resistance to methyl-methane sulfonate. PALF also interacts directly with KU86, LIGASEIV and phosphorylated XRCC4 proteins and possesses endo/exonuclease activity at protruding DNA ends. Various treatments that produce double-strand breaks induce formation of PALF foci, which fully coincide with gammaH2AX foci. Thus, PALF and the CYR motif may play important roles in DNA repair of higher eukaryotes. PMID- 17396151 TI - Structural basis for enzymatic excision of N1-methyladenine and N3-methylcytosine from DNA. AB - N(1)-methyladenine (m(1)A) and N(3)-methylcytosine (m(3)C) are major toxic and mutagenic lesions induced by alkylation in single-stranded DNA. In bacteria and mammals, m(1)A and m(3)C were recently shown to be repaired by AlkB-mediated oxidative demethylation, a direct DNA damage reversal mechanism. No AlkB gene homologues have been identified in Archaea. We report that m(1)A and m(3)C are repaired by the AfAlkA base excision repair glycosylase of Archaeoglobus fulgidus, suggesting a different repair mechanism for these lesions in the third domain of life. In addition, AfAlkA was found to effect a robust excision of 1,N(6)-ethenoadenine. We present a high-resolution crystal structure of AfAlkA, which, together with the characterization of several site-directed mutants, forms a molecular rationalization for the newly discovered base excision activity. PMID- 17396152 TI - Nanodiscs unravel the interaction between the SecYEG channel and its cytosolic partner SecA. AB - The translocon is a membrane-embedded protein assembly that catalyzes protein movement across membranes. The core translocon, the SecYEG complex, forms oligomers, but the protein-conducting channel is at the center of the monomer. Defining the properties of the SecYEG protomer is thus crucial to understand the underlying function of oligomerization. We report here the reconstitution of a single SecYEG complex into nano-scale lipid bilayers, termed Nanodiscs. These water-soluble particles allow one to probe the interactions of the SecYEG complex with its cytosolic partner, the SecA dimer, in a membrane-like environment. The results show that the SecYEG complex triggers dissociation of the SecA dimer, associates only with the SecA monomer and suffices to (pre)-activate the SecA ATPase. Acidic lipids surrounding the SecYEG complex also contribute to the binding affinity and activation of SecA, whereas mutations in the largest cytosolic loop of the SecY subunit, known to abolish the translocation reaction, disrupt both the binding and activation of SecA. Altogether, the results define the fundamental contribution of the SecYEG protomer in the translocation subreactions and illustrate the power of nanoscale lipid bilayers in analyzing the dynamics occurring at the membrane. PMID- 17396153 TI - Reconstruction of a pathway of antigen processing and class II MHC peptide capture. AB - Endocytosed antigens are proteolytically processed and small amounts of peptides captured by class II MHC molecules. The details of antigen proteolysis, peptide capture and how destruction of T-cell epitopes is avoided are incompletely understood. Using the tetanus toxin antigen, we show that the introduction of 3-6 cleavage sites is sufficient to trigger a partially unfolded conformation able to bind to class II MHC molecules. The known locations of T-cell epitopes and protease cleavage sites predict that large domains of processed antigen (8-35 kDa) are captured under these conditions. Remarkably, when antigen is bound to the B-cell antigen receptor (BCR), processing can trigger a concerted 'hand-over' reaction whereby BCR-associated processed antigen is captured by neighbouring class II MHC molecules. Early capture of minimally processed antigen and confinement of the processing and class II MHC loading reaction to the membrane plane may improve the likelihood of T-cell epitope survival in the class II MHC pathway and may help explain the reciprocal relationships observed between B- and T-cell epitopes in many protein antigens and autoantigens. PMID- 17396154 TI - A permeable cuticle in Arabidopsis leads to a strong resistance to Botrytis cinerea. AB - The plant cuticle composed of cutin, a lipid-derived polyester, and cuticular waxes covers the aerial portions of plants and constitutes a hydrophobic extracellular matrix layer that protects plants against environmental stresses. The botrytis-resistant 1 (bre1) mutant of Arabidopsis reveals that a permeable cuticle does not facilitate the entry of fungal pathogens in general, but surprisingly causes an arrest of invasion by Botrytis. BRE1 was identified to be long-chain acyl-CoA synthetase2 (LACS2) that has previously been shown to be involved in cuticle development and was here found to be essential for cutin biosynthesis. bre1/lacs2 has a five-fold reduction in dicarboxylic acids, the typical monomers of Arabidopsis cutin. Comparison of bre1/lacs2 with the mutants lacerata and hothead revealed that an increased permeability of the cuticle facilitates perception of putative elicitors in potato dextrose broth, leading to the presence of antifungal compound(s) at the surface of Arabidopsis plants that confer resistance to Botrytis and Sclerotinia. Arabidopsis plants with a permeable cuticle have thus an altered perception of their environment and change their physiology accordingly. PMID- 17396155 TI - Crystal structures of autoinhibitory PDZ domain of Tamalin: implications for metabotropic glutamate receptor trafficking regulation. AB - Metabotropic glutamate receptors (mGluRs) function as neuronal G-protein-coupled receptors and this requires efficient membrane targeting through associations with cytoplasmic proteins. However, the molecular mechanism regulating mGluR cell surface trafficking remains unknown. We report here that mGluR trafficking is controlled by the autoregulatory assembly of a scaffold protein Tamalin. In the absence of mGluR, Tamalin self-assembles into autoinhibited conformations, through its PDZ domain and C-terminal intrinsic ligand motif. X-ray crystallographic analyses visualized integral parts of the oligomeric self assemblies of Tamalin, which require not only the novel hydrophobic dimerization interface but also canonical and noncanonical PDZ/ligand autoinhibitory interactions. The mGluR cytoplasmic region can competitively bind to Tamalin at a higher concentration, disrupting weak inhibitory interactions. The atomic view of mGluR association suggests that this rearrangement is dominated by electrostatic attraction and repulsion. We also observed in mammalian cells that the association liberates the intrinsic ligand toward a motor protein receptor, thereby facilitating mGluR cell-surface trafficking. Our study suggests a novel regulatory mechanism of the PDZ domain, by which Tamalin switches between the trafficking-inhibited and -active forms depending on mGluR association. PMID- 17396156 TI - Removal of nonnative fish results in population expansion of a declining amphibian (mountain yellow-legged frog, Rana muscosa). AB - The mountain yellow-legged frog (Rana muscosa) was once a common inhabitant of the Sierra Nevada (California, USA), but has declined precipitously during the past century due in part to the introduction of nonnative fish into naturally fishless habitats. The objectives of the current study were to describe (1) the effect of fish removal from three lakes (located in two watersheds) on the small, remnant R. muscosa populations inhabiting those lakes, and (2) the initial development of metapopulation structure in each watershed as R. muscosa from expanding populations in fish-removal lakes dispersed to adjacent habitats. At all three fish-removal lakes, R. muscosa population densities increased significantly following the removal of predatory fish. The magnitude of these increases was significantly greater than that observed over the same time period in R. muscosa populations inhabiting control lakes that remained in their natural fishless condition. Following these population increases, R. muscosa dispersed to adjacent suitable (but unoccupied) sites, moving between 200 and 900 m along streams or across dry land. Together, these results suggest that large-scale removal of introduced fish could result in at least partial reversal of the decline of R. muscosa. Continued monitoring of R. muscosa at the fish-removal sites will be necessary to determine whether the positive effects of fish eradication are sustained over the long-term, especially in light of the increasingly important role played by an emerging infectious disease (chytridiomycosis, caused by Batrachochytrium dendrobatidis) in influencing R. muscosa populations. PMID- 17396157 TI - Multiphoton coherent control in complex systems. AB - Control of multiphoton transitions is demonstrated for a multilevel system by generalizing the instantaneous phase of any chirped pulse as individual terms of a Taylor series expansion. In the case of a simple two-level system, all odd terms in the series lead to population inversion while the even terms lead to self-induced transparency. The results hold for multiphoton transitions that do not have any lower-order photon resonance or any intermediate virtual state dynamics within the laser pulse width. PMID- 17396158 TI - Role of dietary soy protein in obesity. AB - Soy protein is an important component of soybeans and provides an abundant source of dietary protein. Among the dietary proteins, soy protein is considered a complete protein in that it contains ample amounts of all the essential amino acids plus several other macronutrients with a nutritional value roughly equivalent to that of animal protein of high biological value. Soy protein is unique among the plant-based proteins because it is associated with isoflavones, a group of compounds with a variety of biological properties that may potentially benefit human health. An increasing body of literature suggests that soy protein and its isoflavones may have a beneficial role in obesity. Several nutritional intervention studies in animals and humans indicate that consumption of soy protein reduces body weight and fat mass in addition to lowering plasma cholesterol and triglycerides. In animal models of obesity, soy protein ingestion limits or reduces body fat accumulation and improves insulin resistance, the hallmark of human obesity. In obese humans, dietary soy protein also reduces body weight and body fat mass in addition to reducing plasma lipids. Several potential mechanisms whereby soy protein may improve insulin resistance and lower body fat and blood lipids are discussed and include a wide spectrum of biochemical and molecular activities that favorably affect fatty acid metabolism and cholesterol homeostasis. The biologic actions of certain constituents of soy protein, particularly conglycinin, soyasaponins, phospholipids, and isoflavones, that relate to obesity are also discussed. In addition, the potential of soy protein in causing food allergy in humans is briefly discussed. PMID- 17396159 TI - Rasburicase represents a new tool for hyperuricemia in tumor lysis syndrome and in gout. AB - Hyperuricemia is a feature of several pathologies and requires an appropriate and often early treatment, owing to the severe consequences that it may cause. A rapid and massive raise of uric acid, during tumor lysis syndrome (TLS), and also a lower and chronic hyperuricemia, as in gout, mainly damage the kidney. To prevent or treat these consequences, a new therapeutic option is represented by rasburicase, a recombinant form of an enzyme, urate oxidase. This enzyme converts hypoxanthine and xanthine into allantoin, a more soluble molecule, easily cleared by kidney. The several types of urate oxidase have followed each other, with progressive reduction of adverse reactions. The most important among them are allergenicity and the development of antibodies which compromise their effectiveness. Nevertheless, a limit of rasburicase's use remains its cost, which obliges to a judicious choice to prevent TLS in high risk patients with cancer and in case of allergy or impossibility to take allopurinol orally both in TLS and in gout. A large body of evidence confirms the efficacy and safety of rasburicase, even in comparison to the standard drugs used in the aforementioned pathologies. PMID- 17396160 TI - Alterations in arterial blood parameters in patients with liver cirrhosis and ascites. AB - In cirrhotic patients, in addition to hepatocytes and Kuppfer cells dysfunction circulatory anatomic shunt and ventilation/perfusion (V(A)/ Q) ratio abnormalities can induce decrease in partial pressure of oxygen in arterial blood (PaO(2)), in oxygen saturation of hemoglobin (SaO(2)) as well as various acid base disturbances. We studied 49 cases of liver cirrhosis (LC) with ascites compared to 50 normal controls. Causes were: posthepatic 37 (75.51%), alcoholic 7 (14.24%), cardiac 2 (4.08%), and cryptogenic 3 (6.12%). Complications were: upper gastrointestinal bleeding 24 (48.97), hepatic encephalopathy 20 (40.81%), gastritis 28 (57.14%), hepatoma 5 (10.2%), renal hepatic syndrome 2 (4.01%), HbsAg (+) 24 (48.97%), and hepatic pleural effusions 7 (14.28%). Average PaO(2) and SaO(2) were 75.2 mmHg and 94.5 mmHg, respectively, compared to 94.2 mmHg and 97.1 mmHg of the control group, respectively (p value in both PaO(2) and SaO(2 )was p<0.01). Respiratory alkalosis, metabolic alkalosis, metabolic acidosis, respiratory acidosis and metabolic acidosis with respiratory alkalosis were acid base disturbances observed. In conclusion, portopulmonary shunt, intrapulmonary arteriovenous shunt and V(A)/Q inequality can induce a decrease in PaO(2) and SaO(2) as well as various acid-base disturbances. As a result, pulmonary resistance is impaired and patients more likely succumb to infections and adult respiratory distress syndrome. PMID- 17396161 TI - No associations of Helicobacter pylori infection and gastric atrophy with plasma total homocysteine in Japanese. AB - Recent studies have suggested that Helicobacter pylori (H. pylori) infection might be a risk factor for atherosclerosis. Since the bacterium has not been isolated from atherosclerotic lesions, a direct role in atherogenesis is not plausible. We examined associations of plasma total homocysteine (tHcy) and serum folate, independent risk factors for atherosclerosis, with H. pylori infection and subsequent gastric atrophy among 174 patients (78 males and 96 females) aged 20 to 73 years, who visited an H. pylori eradication clinic of Nagoya University from July 2004 to October 2005. Polymorphism genotyping was conducted for methylenetetrahydrofolate reductase (MTHFR) C677T and thymidylate synthase (TS) 28-bp tandem repeats by PCR with confronting two-pair primers and PCR, respectively. H. pylori infection and gastric atrophy were not significantly associated with hyperhomocysteinemia (tHcy > or = 12 nmol/ml), when adjusted by sex, age, smoking, alcohol, and genotypes of MTHFR and TS. The adjusted odds ratio of gastric atrophy for low folate level (< or = 4 mg/ml) was 0.21 (95% confidence interval = 0.05-0.78). The associations of tHcy with serum folate and MTHFR genotype were clearly observed in this dataset. The present study demonstrated that folate and MTHFR genotype were the deterministic factors of plasma tHcy, but not H. pylori infection and subsequent gastric atrophy, indicating that even if H. pylori infection influences the risk of atherosclerosis, the influence may not be through the elevation of homocysteine. PMID- 17396162 TI - Reproductive phase locking of mosquito populations in response to rainfall frequency. AB - The frequency of moderate to heavy rainfall events is projected to change in response to global warming. Here we show that these hydrologic changes may have a profound effect on mosquito population dynamics and rates of mosquito-borne disease transmission. We develop a simple model, which treats the mosquito reproductive cycle as a phase oscillator that responds to rainfall frequency forcing. This model reproduces observed mosquito population dynamics and indicates that mosquito-borne disease transmission can be sensitive to rainfall frequency. These findings indicate that changes to the hydrologic cycle, in particular the frequency of moderate to heavy rainfall events, could have a profound effect on the transmission rates of some mosquito-borne diseases. PMID- 17396163 TI - Mechanisms of copper ion mediated Huntington's disease progression. AB - Huntington's disease (HD) is caused by a dominant polyglutamine expansion within the N-terminus of huntingtin protein and results in oxidative stress, energetic insufficiency and striatal degeneration. Copper and iron are increased in the striata of HD patients, but the role of these metals in HD pathogenesis is unknown. We found, using inductively-coupled-plasma mass spectroscopy, that elevations of copper and iron found in human HD brain are reiterated in the brains of affected HD transgenic mice. Increased brain copper correlated with decreased levels of the copper export protein, amyloid precursor protein. We hypothesized that increased amounts of copper bound to low affinity sites could contribute to pro-oxidant activities and neurodegeneration. We focused on two proteins: huntingtin, because of its centrality to HD, and lactate dehydrogenase (LDH), because of its documented sensitivity to copper, necessity for normoxic brain energy metabolism and evidence for altered lactate metabolism in HD brain. The first 171 amino acids of wild-type huntingtin, and its glutamine expanded mutant form, interacted with copper, but not iron. N171 reduced Cu(2+)in vitro in a 1:1 copper:protein stoichiometry indicating that this fragment is very redox active. Further, copper promoted and metal chelation inhibited aggregation of cell-free huntingtin. We found decreased LDH activity, but not protein, and increased lactate levels in HD transgenic mouse brain. The LDH inhibitor oxamate resulted in neurodegeneration when delivered intra-striatially to healthy mice, indicating that LDH inhibition is relevant to neurodegeneration in HD. Our findings support a role of pro-oxidant copper-protein interactions in HD progression and offer a novel target for pharmacotherapeutics. PMID- 17396164 TI - Probabilistic protein function prediction from heterogeneous genome-wide data. AB - Dramatic improvements in high throughput sequencing technologies have led to a staggering growth in the number of predicted genes. However, a large fraction of these newly discovered genes do not have a functional assignment. Fortunately, a variety of novel high-throughput genome-wide functional screening technologies provide important clues that shed light on gene function. The integration of heterogeneous data to predict protein function has been shown to improve the accuracy of automated gene annotation systems. In this paper, we propose and evaluate a probabilistic approach for protein function prediction that integrates protein-protein interaction (PPI) data, gene expression data, protein motif information, mutant phenotype data, and protein localization data. First, functional linkage graphs are constructed from PPI data and gene expression data, in which an edge between nodes (proteins) represents evidence for functional similarity. The assumption here is that graph neighbors are more likely to share protein function, compared to proteins that are not neighbors. The functional linkage graph model is then used in concert with protein domain, mutant phenotype and protein localization data to produce a functional prediction. Our method is applied to the functional prediction of Saccharomyces cerevisiae genes, using Gene Ontology (GO) terms as the basis of our annotation. In a cross validation study we show that the integrated model increases recall by 18%, compared to using PPI data alone at the 50% precision. We also show that the integrated predictor is significantly better than each individual predictor. However, the observed improvement vs. PPI depends on both the new source of data and the functional category to be predicted. Surprisingly, in some contexts integration hurts overall prediction accuracy. Lastly, we provide a comprehensive assignment of putative GO terms to 463 proteins that currently have no assigned function. PMID- 17396165 TI - Protection of visual functions by human neural progenitors in a rat model of retinal disease. AB - BACKGROUND: A promising clinical application for stem and progenitor cell transplantation is in rescue therapy for degenerative diseases. This strategy seeks to preserve rather than restore host tissue function by taking advantage of unique properties often displayed by these versatile cells. In studies using different neurodegenerative disease models, transplanted human neural progenitor cells (hNPC) protected dying host neurons within both the brain and spinal cord. Based on these reports, we explored the potential of hNPC transplantation to rescue visual function in an animal model of retinal degeneration, the Royal College of Surgeons rat. METHODOLOGY/PRINCIPAL FINDINGS: Animals received unilateral subretinal injections of hNPC or medium alone at an age preceding major photoreceptor loss. Principal outcomes were quantified using electroretinography, visual acuity measurements and luminance threshold recordings from the superior colliculus. At 90-100 days postnatal, a time point when untreated rats exhibit little or no retinal or visual function, hNPC-treated eyes retained substantial retinal electrical activity and visual field with near normal visual acuity. Functional efficacy was further enhanced when hNPC were genetically engineered to secrete glial cell line-derived neurotrophic factor. Histological examination at 150 days postnatal showed hNPC had formed a nearly continuous pigmented layer between the neural retina and retinal pigment epithelium, as well as distributed within the inner retina. A concomitant preservation of host cone photoreceptors was also observed. CONCLUSIONS/SIGNIFICANCE: Wild type and genetically modified human neural progenitor cells survive for prolonged periods, migrate extensively, secrete growth factors and rescue visual functions following subretinal transplantation in the Royal College of Surgeons rat. These results underscore the potential therapeutic utility of hNPC in the treatment of retinal degenerative diseases and suggest potential mechanisms underlying their effect in vivo. PMID- 17396166 TI - Recent Progress in the Treatment of Advanced Prostate Cancer With Intermittent Dose-Intense Calcitriol (DN-101). AB - Docetaxel is becoming standard therapy for androgen-independent prostate cancer (AIPC), and investigational agents are being added to docetaxel to assess potential additive effects and synergy. Although one of these agents, calcitriol, has repeatedly demonstrated antiproliferative properties against cancer of the prostate, breast, colon, and lung, the antineoplastic activity of calcitriol requires superphysiologic levels. Unfortunately, chronic exposure to superphysiologic levels of calcitriol causes hypercalcemia and resulting toxicity. Therefore, a host of analogues of calcitriol have been investigated for antineoplastic function, including intermittent dose-intense calcitriol, or DN 101. Because of encouraging results from phase II studies of DN-101 combined with docetaxel, the ASCENT (AIPC Study of Calcitriol Enhancement of Taxotere) phase II trial investigated docetaxel plus DN-101 versus docetaxel plus placebo in 250 men with metastatic AIPC and an abnormal baseline prostate-specific antigen (PSA) level. Although the ASCENT trial did not achieve its primary endpoint for increased PSA response, there was a significant trend in PSA response rate in the DN-101 arm. DN-101 in combination with docetaxel seems to improve overall survival and, interestingly, has a favorable safety profile compared with docetaxel alone. The DN-101/docetaxel combination is currently being studied in a much larger international trial, ASCENT-2. PMID- 17396167 TI - Diagnosis of interstitial cystitis/ painful bladder syndrome in patients with overactive bladder symptoms. AB - Overactive bladder (OAB) and interstitial cystitis (IC) have similar symptoms, including urinary urgency/frequency and nocturia, making them difficult to differentiate on the basis of clinical presentation alone. Both conditions may represent a clinical manifestation of a hypersensitive bladder and should be included in the differential diagnosis for patients who present with urgency/ frequency. It is especially important that IC be considered in patients with OAB that is refractory to treatment. The proposed diagnostic framework may be useful for differentiating IC from OAB and for facilitating appropriate treatment. PMID- 17396168 TI - Uric Acid nephrolithiasis: recent progress and future directions. AB - The prevalence of urolithiasis has been increasing for the past few decades in industrialized nations. Uric acid calculi account for a significant percentage of urinary stones. Certain risk factors may be involved in the pathogenesis of uric acid nephrolithiasis, including hyperuricosuria, low urinary volume, and persistently low urinary pH. Patients with medical conditions that promote profound hyperuricosuria are at high risk of developing uric acid calculi. These conditions include chronic diarrheal states; myeloproliferative disorders; insulin resistance, including diabetes mellitus; and monogenic metabolic disorders, such as Lesch-Nyhan syndrome. Computed tomography can provide a definitive diagnosis. Except in cases in which there is severe obstruction, progressive azotemia, serious infection, or unremitting pain, the initial treatment of patients with uric acid nephrolithiasis should be medical dissolution therapy because this approach is successful in the majority of cases. A thorough review of the epidemiology and pathophysiology of uric acid nephrolithiasis is crucial for the diagnosis, treatment, and prevention of stones in patients with this condition. PMID- 17396170 TI - Updates in Pediatric Urology: Highlights of the American Academy of Pediatrics Section on Urology Annual Meeting, October 7-9, 2006, Atlanta, GA. PMID- 17396169 TI - Is the testis a chemo-privileged site? Is there a blood-testis barrier? AB - The incidence of testicular cancer, primarily seminoma, has been increasing in many countries, including the United States. The testis is often the site of residual cancer after adequate treatment with systemic chemotherapy. The blood testis barrier is commonly cited as the explanation for residual tumor within the gonad after chemotherapy and as the indication for delayed orchiectomy. Conversely, complete eradication of viable tumor from the primary site is common and argues against the testis as a "tumor sanctuary." Residual tumor is also demonstrated within metastatic foci, and the disparity between the histopathologic response of the primary tumor and metastatic sites may be best explained by tumor heterogeneity and multiple tumor clones. Regardless of the scientific and academic arguments, delayed radical orchiectomy remains an important part of treatment for patients undergoing primary chemotherapy. PMID- 17396171 TI - Solitary fibrous tumor of the kidney: a case report and review of the literature. AB - A solitary fibrous tumor (SFT) is an unusual spindle cell neoplasm that usually occurs in the pleura but has recently been described in diverse extrapleural sites. Urogenital localization is rare, and only 19 cases of SFT of the kidney have been described. We report a case of a large SFT clinically thought to be renal cell carcinoma arising in the kidney of a 70-year-old man. The tumor was well circumscribed and composed of a mixture of spindle cells and dense collagenous bands, with areas of necrosis or cystic changes noted macroscopically and microscopically. Immunohistochemical studies revealed reactivity for CD34, CD99, and Bcl-2 protein, with no staining for keratin, S-100 protein, or muscle markers, confirming the diagnosis of SFT. This tumor is benign in up to 90% of cases. The immunohistochemical study is the key to diagnosis. PMID- 17396172 TI - Postprostatectomy risk stratification based on detection of occult lymph node metastasis. PMID- 17396173 TI - Dextranomer/Hyaluronic Acid implantation for vesicoureteral reflux. PMID- 17396174 TI - Botulinum toxin a in the treatment of neurogenic and idiopathic urinary incontinence. PMID- 17396175 TI - Structural and functional properties of metarhodopsin III: recent spectroscopic studies on deactivation pathways of rhodopsin. AB - The activation of rhodopsin has been the focus of researchers over the past decades, revealing many aspects of the activation pathways of this prototypical G protein-coupled receptor on a molecular level, starting with the light-dependent isomerization of its retinal chromophore from 11-cis to all-trans and leading eventually to the large scale helix movements in the transition to the active receptor state, Meta II. Comparatively little is known, however, on the deactivation pathways of the light receptor, which represent essential steps in maintaining a functional photoreceptor cell. Rhodopsin's active receptor species, Meta II, decays by two fundamentally different pathways, either forming the apoprotein opsin by release of the activating all-trans retinal ligand from its binding pocket, or by a thermal isomerization of this ligand to a less activating species in the transition to metarhodopsin III (Meta III). Both decay products, opsin and Meta III, are largely inactive under physiological conditions, yet they do not restore the complete inactivity of the dark state. Although some properties of Meta III have been described already in the 1960s, its molecular nature and the pathways of its formation have remained rather obscure. In this review, we focus on recent studies from our laboratories, which have provided a major progress in our understanding of the Meta III deactivation pathway and its potential physiological roles. Using Fourier-transform infrared (FTIR) difference spectroscopy in combination with a variety of other spectroscopic and biochemical techniques and quantum chemical calculations, we have developed a general picture of the interplay between the retinal ligand and the receptor protein, which is compared to similar reaction mechanisms in invertebrate photoreceptors and microbial retinal proteins. PMID- 17396176 TI - Helices and Sheets in vacuo. AB - The structures and properties of unsolvated peptides large enough to possess secondary structure have been examined by experiments and simulations. Some of the factors that stabilize unsolvated helices and sheets have been identified. The charge, in particular, plays a critical role in stabilizing alpha-helices and destabilizing beta-sheets. Some helices are much more stable in vacuum than in aqueous solution. Factors like helix propensity, context, and the incorporation of specific stabilizing interactions have been examined. The helix propensities in vacuum differ from those found in solution. Studies of the hydration of unsolvated peptides can be performed one water molecule at a time. The first few water molecules only bind weakly to unsolvated peptides, and they bind much more strongly to some conformations than to others. The most favorable binding locations are not the protonation sites, but clefts or pockets where a water molecule can establish a network of hydrogen bonds. Non-covalent interactions between secondary structure elements leads to the formation of tertiary structure. Helical peptides assemble into complexes with a variety of intriguing structures. The intramolecular coupling of helices to make antiparallel coiled coil geometries has also been investigated with model peptides. PMID- 17396177 TI - Electronic coupling in the excited electronic state of stacked DNA base homodimers. AB - The nature of the electronic coupling of stacked nucleic acid bases adenine (A), thymine (T), and cytosine (C), in A-A, T-T, and C-C complexes in their excited states was investigated; a different character of the electronic coupling for the T-T complex was shown. PMID- 17396178 TI - Dissymmetry effects on the laser spectroscopy of supersonically expanded rare gas/chiral arene heteroclusters. AB - The R2PI-TOF spectra of supersonically expanded rare gas/chiral arene heteroclusters have been rationalized in terms of the distortion of the pi electron density reflecting the different dipole and quadrupole momenta induced in the rare gas atoms by interaction with the opposite pi-faces of the chiral arene itself. PMID- 17396179 TI - How accurate is the density functional theory combined with symmetry-adapted perturbation theory approach for CH-pi and pi-pi interactions? A comparison to supermolecular calculations for the acetylene-benzene dimer. AB - Five different orientations of the acetylene-benzene dimer including the T-shaped global minimum structure are used to assess the accuracy of the density functional theory combined with symmetry adapted perturbation theory (DFT-SAPT) approach in its density-fitting implementation (DF-DFT-SAPT) for the study of CH pi and pi-pi interactions. The results are compared with the outcome of counterpoise corrected supermolecular calculations employing second-order Moller Plesset (MP2), spin-component scaled MP2 (SCS-MP2) and single and double excitation coupled cluster theory including perturbative triple excitations (CCSD(T)). For all considered orientations MP2 predicts much deeper potential energy curves with considerably shifted minima compared to CCSD(T) and DFT-SAPT. In spite of being an improvement over the results of MP2, SCS-MP2 tends to underestimate the well depth while DFT-SAPT, employing an asymptotically corrected hybrid exchange-correlation potential in conjunction with the adiabatic local density approximation for the exchange-correlation kernel, is found to be in excellent agreement with CCSD(T). Furthermore, DFT-SAPT provides a detailed understanding of the importance of the electrostatic, induction and dispersion contributions to the total interaction energy and their repulsive exchange corrections. PMID- 17396180 TI - Mode-specific energy absorption by solvent molecules during CO2 vibrational cooling. AB - Non-equilibrium molecular dynamics (NEMD) simulations of energy transfer from vibrationally excited CO(2) to CCl(4) and CH(2)Cl(2) solvent molecules are performed to identify the efficiency of different energy pathways into the solvent bath. Studying in detail the work performed by the vibrationally excited solute on the different solvent degrees of freedom, it is shown that vibration-to vibration (V-V) processes are strongly dominant and controlled by those accepting modes which are close in frequency to the CO(2) bend and symmetric stretch vibration. PMID- 17396181 TI - A strategy for predicting the crystal structures of flexible molecules: the polymorphism of phenobarbital. AB - A computational exploration of the low energy crystal structures of the pharmaceutical molecule phenobarbital is presented as a test of an approach for the crystal structure prediction of flexible molecules. Traditional transferable force field methods of modelling flexible molecules are unreliable for the level of accuracy required in crystal structure prediction and we outline a strategy for improving the evaluation of relative energies of large sets of crystal structures. The approach involves treating the molecule as a set of linked rigid units, whose conformational energy is expressed as a function of the relative orientations of the rigid groups. The conformational energy is calculated by electronic structure methods and the intermolecular interactions using an atomic multipole description of electrostatics. A key consideration in our approach is the scalability to more typical pharmaceutical molecules of higher molecular weight with many more atoms and degrees of flexibility. Based on our calculations, crystal structures are proposed for the as-yet uncharacterised forms IV and V, as well as further polymorphs of phenobarbital. PMID- 17396182 TI - Molecular ordering in a biaxial smectic-A phase studied by scanning transmission X-ray microscopy (STXM). AB - Results of STXM investigations of a binary mixture (-TNF = 2 : 1; SmA(b) 140 M 180 Iso) known to form a SmA(b) phase [T. Hegmann, J. Kain, S. Diele, G. Pelzl and C. Tschierske, Angew. Chem. Int. Ed., 2001, 40, 887] are presented. Near edge X-ray absorption fine spectra (NEXAFS) of the -TNF board-like aggregates, in particular the intensity of the low energy peaks associated with aromatic ring pi* orbitals (284.5-286.5 eV), show that the molecular plane of these aggregates is very sensitive to the relative orientation of electric field vector E of linearly polarized light, which is used to determine the molecular orientation in the LC phase. The observed strong in-plane dichroic signal suggests the predominant orientation of the -TNF aggregates to be along the smectic layer normal as well as long-range ordering of the in-plane molecular orientation (biaxiality). Orientational maps derived from series of measurements at different sample rotation angles around the specimen normal clearly show a Schlieren-type texture, and permit a detailed examination of exclusive +/-(1/2) disclination theoretically predicted for the SmA(b) phase. PMID- 17396183 TI - On the use of electrokinetic phenomena of the second kind for probing electrode kinetic properties of modified electron-conducting surfaces. AB - The electrokinetic features of electron-conducting substrates, as measured in a conventional thin-layer electrokinetic cell, strongly depend on the extent of bipolar faradaic depolarisation of the interface formed with the adjacent electrolytic solution. Streaming potential versus applied pressure data obtained for metallic substrates must generally be interpreted on the basis of a modified Helmholtz-Smoluchowski equation corrected by an electronic conduction term-non linear with respect to the lateral potential and applied pressure gradient-that stems from the bipolar electrodic behavior of the metallic surface. In the current study, streaming potential measurements have been performed in KNO(3) solutions on porous plugs made of electron-conducting grains of pyrite (FeS(2)) covered by humic acids. For zero coverage, the extensive bipolar electronic conduction taking place in the plug-depolarized by concomitant and spatially distributed oxidation and reduction reactions of Fe(2+) and Fe(3+) species-leads to the complete extinction of the streaming potential over the entire range of applied pressure examined. For low to intermediate coverage, the local electron transfer kinetics on the covered regions of the plug becomes more sluggish. The overall bipolar electronic conduction is then diminished which leads to an increase in the streaming potential with a non-linear dependence on the pressure. For significant coverage, a linear response is observed which basically reflects the interfacial double layer properties of the humics surface layer. A tractable, semi-analytical model is presented that reproduces the electrokinetic peculiarities of the complex and composite system FeS(2)/humics investigated. The study demonstrates that the streaming potential technique is a fast and valuable tool for establishing how well the electron transfer kinetics at a partially or completely depolarised bare electron-conducting substrate/electrolyte solution interface is either promoted (catalysis) or blocked (passivation) by the presence of a discontinuous surface layer. PMID- 17396184 TI - Dissociative electron attachment to abasic DNA. AB - Thin films of the short single DNA strand, GCAT, in which one of the bases has been removed were bombarded with 3 to 15 eV electrons. The yield functions of the H(-), O(-) and OH(-) ions desorbed from these films exhibit a broad peak near 9 eV, which is attributed to dissociative electron attachment to the basic molecules. Whereas removal of any one of the bases considerably decreases N glycosidic and backbone C-O bond scission, the creation of basic sites does not appreciably modify bond rupture leading to anion electron stimulated desorption. These seemingly contradictory results make it possible to propose a detailed mechanism leading to the transfer of electrons in the range 5-13 eV within DNA. PMID- 17396185 TI - Size-dependent structural transformations of hematite nanoparticles. 1. Phase transition. AB - Using Fourier Transform InfraRed (FTIR) spectroscopy, Raman spectroscopy, X-ray diffraction (XRD), and Transmission Electron Microscopy (TEM), we characterize the structure and/or morphology of hematite (alpha-Fe(2)O(3)) particles with sizes of 7, 18, 39 and 120 nm. It is found that these nanoparticles possess maghemite (gamma-Fe(2)O(3))-like defects in the near surface regions, to which a vibrational mode at 690 cm(-1), active both in FTIR and Raman spectra, is assigned. The fraction of the maghemite-like defects and the net lattice disorder are inversely related to the particle size. However, the effect is opposite for nanoparticles grown by sintering of smaller hematite precursors under conditions when the formation of a uniform hematite-like structure throughout the aggregate is restricted by kinetic issues. This means that not only particle size but also the growth kinetics determines the structure of the nanoparticles. The observed structural changes are interpreted as size-induced alpha-Fe(2)O(3)<-->gamma Fe(2)O(3) phase transitions. We develop a general model that considers spinel defects and absorbed/adsorbed species (in our case, hydroxyls) as dominant controls on structural changes with particle size in hematite nanoparticles, including solid-state phase transitions. These changes are represented by trajectories in a phase diagram built in three phase coordinates-concentrations of spinel defects, absorbed impurities, and adsorbed species. The critical size for the onset of the alpha-->gamma phase transition depends on the particle environment, and for the dry particles used in this study is about 40 nm. The model supports the existence of intermediate phases (protohematite and hydrohematite) during dehydration of goethite. We also demonstrate that the hematite structure is significantly less defective when the nanoparticles are immersed in water or KBr matrix, which is explained by the effects of the electrochemical double layer and increased rigidity of the particle environment. Finally, we revise the problem of applicability of IR spectroscopy to the lattice vibrations of hematite nanoparticles, demonstrating that structural comparison of different samples is much more reliable if it is based on the E(u) band at about 460 cm(-1) and the spinel band at 690 cm(-1), instead of the A(2u)/E(u) band at about 550 cm(-1) used in previous work. The new methodology is applied to analysis of the reported IR spectra of Martian hematite. PMID- 17396186 TI - In-hospital and late outcomes after coronary stenting in patient with unstable angina and myelodysplastic syndrome. AB - Sixty-one-year-old male patient with diagnosis of myelodysplastic syndrome and unstable angina was submitted to coronary angiography and implant of stent. His Blood cell count revealed 40,000 platelets/mm3. Coronary angiography with previous platelet transfusion showed obstruction of 80% of the right coronary artery (RCA). Following the administration of clopidogrel, the patient was submitted to another platelet transfusion and stent implantation in the RCA lesion. No bleeding was observed after the introducers removal. After 6 months, treadmill test was positive and new coronary aniography, in the same conditions, showed in-stent restenosis. This case report suggests that coronary stent implantation in patients with thrombocytopenia is a safe procedure, provided that prophylactic platelet transfusion is performed, although late restenosis may occur. PMID- 17396187 TI - Surgical correction of total anomalous pulmonary venous drainage in an adult. AB - Total anomalous pulmonary venous drainage (TAPVD) is rarely seen in adults, because this congenital heart disease almost always requires surgical treatment in the neonatal period, often on an emergency basis. We report a patient that, despite being diagnosed during childhood, underwent surgical repair at age 25, about one year after his clinical condition worsened. PMID- 17396188 TI - Angina pectoris in patient with hyperthyroidism and normal angiography coronary. AB - In the presence of angina pectoris in a premenopausal woman without significant risk factors for coronary disease, we have to rule out other causes of coronary lesion of non atherosclerotic origin. The relations between hyperthyroidism and the cardiovascular system are well known, but hyperthyroidism is responsable for less than 5% of all causes of chest pain. We present a clinical case of a 47 year old woman with typical chest pain and eletrocardiogram (EKG) suggesting coronary ischemia but with normal laboratory data. Anamnesis, clinical and the laboratory data confirmed the diagnosis of hyperthyroidism. Further investigation showed a normal coronary angiography. After treatment with radioiodine and the establishment of euthyroidism, the patient remained asymptomatic and EKG and myocardial scintilography were negative for ischemia. These results suggest a cause and effect relationship between thyroid overactivity and myocardial ischemia, implying a probable etiological role for hyperthyroidism in the clinical and EKG findings. PMID- 17396189 TI - Myocardial infarction associated with the use of a dietary supplement rich in ephedrine in a young athlete. AB - Dietary supplements containing ephedrine and other alkaloids related to ephedrine are largely consumed in various countries, with the purpose of energetic stimulation and weight loss. Despite the fact that it is not approved for marketing in Brazil, these products may be freely purchased over the Internet or at gyms/fitness centers. We report the case of a young athlete with no risk factors for cardiovascular disease who experienced a myocardial infarction during the period in which he used an ephedrine-rich supplement. PMID- 17396190 TI - Multivessel percutaneous treatment of Takayasu arteritis. AB - We report the case of a female patient with obstructive lesions in the right and left carotid, right renal, left subclavian and left common iliac arteries which were percutaneously treated. PMID- 17396191 TI - Cardiac syncope induced by glossopharyngeal "neuralgia": a rare presentation. AB - The first description of severe pain in the distribution of the glossopharyngeal nerve is credited to Weisenberg, in 1910, in a patient with cerebellopontine angle tumor. However, it was Harris, in 1926, who coined the term glossopharyngeal neuralgia to describe this rare condition characterized by paroxysms of excruciating pain located laterally at the back of the tongue, soft palate, throat, and lateral and posterior pharynx, radiating to the ear. Swallowing, coughing, yawning or chewing may trigger pain, which usually lasts from seconds to minutes. The association between glossopharyngeal neuralgia and syncope is very rare, being identified by brief episodes of bradycardia, asystole, and hypotension. Such an association, with this same pathophysiology, was first described by Riley et al in 1942. PMID- 17396192 TI - Tilt table test in the differential diagnosis of refractory "epilepsy". AB - Epilepsy is one of the most frequent causes of neurological disorders in young adults. We report the case of a patient who lived with the diagnosis of refractory epilepsy for twelve years, when actually the cause of the symptoms could be found with the performance of a tilt table test. Cardiologists should be aware of the possible diagnosis of neurocardiogenic syncope in patients previously diagnosed with epilepsy, especially in those with difficult therapeutic control. PMID- 17396193 TI - Tadalafil as treatment for idiopathic pulmonary arterial hypertension. AB - Phosphodiesterase inhibitors like sildenafil have already been shown to improve functional capacity and hemodynamics in the treatment of pulmonary arterial hypertension. Few studies address the effects of new phosphodiesterase inhibitors as tadalafil. We report a case of a patient with idiopathic pulmonary arterial hypertension in functional class IV (New York Heart Association) with significant response to treatment with tadalafil. PMID- 17396194 TI - Aortic valve rupture due to blunt chest trauma. AB - We report a patient admitted to our hospital with aortic valve rupture due to blunt chest trauma. The aortic rupture was accurately identified by the transesophageal echocardiogram, allowing a better surgical approach. PMID- 17396195 TI - The International Liaison Committee on Resuscitation (ILCOR). Roll in guidelines 2005-2010 for cardiopulmonary resuscitation and emergency cardiovascular care. PMID- 17396196 TI - How long...? Possibilities, limits, criteria. PMID- 17396197 TI - Magnetic resonance is useful in heart valvular disease. PMID- 17396198 TI - Saddle shape of mitral valve annulus: three-dimensional transthoracic echocardiography. PMID- 17396199 TI - Progression of ventricular repolarization in left bundle branch block in non compaction of the myocardium. PMID- 17396200 TI - Electrocardiographic abnormalities preceding ventricular premature beats. PMID- 17396201 TI - Cilostazol, a phosphodiesterase III inhibitor: future perspectives in atherosclerosis. PMID- 17396202 TI - The role of aerobic exercise on endothelial function and on cardiovascular risk factors. PMID- 17396203 TI - Application of the stratum-specific likelihood ratio (SSLR) analysis to results of a depressive symptoms screening survey among Japanese workers. AB - BACKGROUND: The stratum-specific likelihood ratio (SSLR) has been widely used as a convenient method to interpret the results of screening surveys. In the present paper, an SSLR analysis is applied to the results of a survey that used the Center for Epidemiologic Studies Depression Scale (CES-D) to identify depressive symptoms in a group of Japanese workers in an attempt to determine score categories that have predictive clinical values in a screening context. METHODS: The self-reported CES-D and the Mini International Neuropsychiatric Interview (MINI) were administered to 2,219 workers (84.2% men; age 21-68 years) during a periodical medical examination. The SSLR was calculated to determine strata with optimal discrimination for major depressive disorder (MDD). RESULTS: For MDD, the SSLR was 0.06 (95%CI: 0.02-0.18) for the CES-D score range 0-16, 1.90 (0.78-4.62) for the score range 17-19, and 12.4 (10.2-15.1) for the score above 20. CONCLUSIONS: The SSLR shown in this study proposes score categories that guide clinicians in selecting the workers most at risks of having MDD among Japanese workers. PMID- 17396204 TI - Involuntary commitment in psychiatric care: what drives the decision? AB - BACKGROUND: Psychiatric commitment laws have been reformed in many European countries. We assessed the relative importance of the different legal criteria in explaining involuntary commitment under the Belgian Mental Health Act of 1990. METHOD: Psychiatric assessments were requested for 346 patients living in Brussels who were randomly selected from a larger group and were being considered for involuntary commitment. A retrospective study of these patients' files was carried out. RESULTS: More than half of the requests for involuntary commitment were turned down. The lack of a less restrictive alternative form of care was the criterion most crucial in decisions in favour of commitment. Alternative forms of care were more likely to be unavailable for psychotic individuals, foreigners, and patients not living in a private household. CONCLUSION: Involuntary commitment is mainly due to the inability of the mental health care system to provide more demanding patients with alternative forms of care. PMID- 17396205 TI - Global and relationship-specific perceptions of support and the development of postpartum depressive symptomatology. AB - BACKGROUND: A lack of social support has consistently been demonstrated to be an important modifiable risk factor for postpartum depression. As such, a greater understanding of specific support variables may assist health professionals in the development of effective preventive interventions. The purpose of this paper was two-fold: (1) to determine if women discriminated between global and relationship-specific perceptions of support, and (2) to examine the influence of global and relationship-specific perceptions of support in the immediate postpartum period on the development of depressive symptomatology at 8 weeks postpartum. METHODS: As part of a longitudinal study, a diverse sample of 594 mothers completed questionnaires that included the Edinburgh Postnatal Depression Scale (EPDS) and global and relationship-specific (e.g., partner, mother, and other women with children) measures of support. RESULTS: Mothers clearly discriminated between global and relationship-specific perceptions of support and those with depressive symptomatology at 8 weeks had significantly lower perceptions of both global and relationship-specific support at 1-week postpartum. Using discriminant function analysis, four variables, reliable reliance from partner, nurturance from partner, attachment to other women with children, and EPDS score at 1-week postpartum, differentiated between mothers who experienced depressive symptomatology at 8 weeks and those who did not. CONCLUSION: Relationship-specific interventions may be beneficial if they include strategies that target a positive partner relationship through preceptions of reliable alliance and feeling needed and provide opportunites for interaction with other mothers. Maternal mood at 1 week postpartum was the largest predictor of depressive symptomatology at 8 weeks. PMID- 17396206 TI - The "continence gap": a critical concept in obstetric fistula repair. PMID- 17396207 TI - Tension-free vaginal tape: poor intraoperative cough test as a predictor of postoperative urinary retention. AB - The purpose of this study was to determine if the quality of the intraoperative cough test could help to predict which patient would fail the post void residual test (PVR) immediately after a tension-free vaginal tape (TVT) procedure. Patients undergoing a TVT procedure only, under spinal or local anesthesia were enrolled. Patients were divided into two groups based on the outcome of the first postoperative PVR, failure group (FG) vs successful group (SG). Before adjusting the tape, patients underwent a standardized cough test. The quality of the cough test was determined to be either good or poor based on whether every cough produced a spurt of urine or not. Variables analyzed between the FG and SG were demographic and urodynamic data. Multivariate logistic regression analysis was used to calculate the adjusted odds ratios. Twenty-six (60.5%) women passed and 17 (39.5%) failed the initial postoperative PVR evaluation. There was a 4.89-fold greater odds of failing the postoperative PVR for women 65 and older compared to younger women (OR 4.89, 95% CI [1.07-26.45]). In addition, there was an 8.63-fold greater odds of failing postoperative PVR for patients with poor quality cough test (OR 8.63, 95% CI [1.54-54.66]). However, multivariate logistic regression analysis revealed that poor quality cough test was the only significant predictor for failing a postoperative PVR (OR 6.83, 95% CI [1.39-33.49], P = 0.018). A poor quality intraoperative cough test at the time of TVT procedure is a predictor of immediate postoperative urinary retention. PMID- 17396208 TI - Changes of collagen type III and decorin in paraurethral connective tissue from women with stress urinary incontinence and prolapse. AB - Thirteen premenopausal women with stress urinary incontinence (SUI), 6 with SUI and prolapse, 9 with prolapse, and 19 without prolapse were enrolled to observe the content change of collagen type III and the expression of decorin mRNA in paraurethral connective tissues. Collagen type III from transvaginal biopsies was assayed by immunohistochemical staining and decorin mRNA was detected by real time PCR. Premenopausal women with SUI had a significantly decreased level of collagen type III. Decorin mRNA expression was significantly increased in both premenopausal SUI+prolapse group and premenopausal prolapse group reflected by the decrease of DeltaCt value compared to their corresponding controls. The results suggest that a high level of decorin mRNA might be associated with the reduced content of collagen type III, resulting in a less flexible form of extracellular matrix in the connective tissue in SUI and prolapse patients. PMID- 17396209 TI - Adiponectin is a predictor of bone mineral density in middle-aged premenopausal women. AB - Adipose-modulated biochemical signal that explains some of the association between fat mass and bone mineral density (BMD) is adiponectin. The results demonstrated an independent association between adiponectin and BMD, while the influence of adiponectin on bone mineral content is mediated by fat free mass in middle-aged women. INTRODUCTION: Positive association between fat mass (FM) and bone mineral density (BMD) is mediated by biochemical factors. METHODS: The relationship between plasma adiponectin concentration and BMD in 98 sedentary premenopausal women aged 38-49 years with a body mass index range of 20.0-42.1 kg/m(2) was examined. Different body composition and blood biochemical parameters were measured to adjust for possible confounding variables. RESULTS: The association between adiponectin and BMD values (total BMD: beta = -0.919; p = 0.0001, femoral neck BMD: beta = -0.925; p = 0.0001 and lumbar spine BMD: ss = 0.912; p = 0.0001) was independent of the influences that measured body composition, hormonal and insulin resistance factors may exert on BMD (p < 0.02). However, adiponectin explained only 3-12% of the variations in measured BMD variables. Similarly, adiponectin was associated with total bone mineral content (BMC; beta = -0.911; p = 0.0001) and remained associated in different analyses that controlled for possible confounding parameters (p < 0.01). However, the association between adiponectin and total BMC was no longer significant when adjusted for fat free mass (FFM; p > 0.21). CONCLUSIONS: Adiponectin is an independent predictor of BMD, while its independent contribution to the interindividual variance in measured values is only modest. The influence of adiponectin on total BMC is mediated or confounded by FFM in middle-aged premenopausal women. PMID- 17396210 TI - Automated solid phase extraction and quantitative measurement of 2,3-dibromo-1 propanol in urine using gas chromatography-mass spectrometry. AB - 2,3-Dibromo-1-propanol (DBP) was used as an active flame retardant in the 1970s. It was also used as an intermediate in the preparation of insecticide formulations, pharmaceuticals and the flame retardants tris(2,3-dibromopropyl) phosphate (Tris-BP) and tetrabromobisphenol A bis (2,3-dibromopropyl ether). DBP is also produced in vivo as a metabolic product of Tris-BP in humans. In 1977, sleepwear containing DBP and Tri-BP was banned because of evidence of carcinogenicity animal studies. Although the production of DBP was reduced after 1977, studies show that DBP is still detected in indoor air and dust; hence, the U.S. population may be exposed potentially to DBP. Only a few methods have been reported in the literature for assessing exposure to DBP or Tris-BP by measuring DBP in urine. These methods are based on a labor-intensive and time-consuming liquid-liquid extraction for the isolation of DBP from the urine matrix. To measure urinary DBP in humans, a fast, accurate, and sensitive method was developed with a limit of detection of 0.1 ng/mL and extraction recovery of 96%. This method involves enzymatic cleavage of the DBP-glucuronide or sulfate conjugate, automated solid phase extraction, and analysis by gas chromatography mass spectrometry using 1,4-dibromo-2-butanol as the internal standard. PMID- 17396211 TI - Multiple cytokine and acute-phase protein gene transcription in West Greenland sledge dogs (Canis familiaris) dietary exposed to organic environmental pollutants. AB - Exposure levels of persistent organic pollutants, such as polychlorinated biphenyls and DDTs, are high in Arctic apex predators and Inuit peoples and are suspected to have negative impacts on their immune systems. We conducted a controlled generational study on liver tissue and ethylenediaminetetraacetic acid blood cytokine and acute-phase protein (APP) mRNA expressions using reverse transcriptionase-polymerase chain reaction in West Greenland sledge dogs (Canis familiaris); contaminated minke whale (Balaenoptera acutorostrata) blubber serves as dietary pollutant source. Two of seven blood cytokine expressions (IL-6 and IL 12) and three of five APP expressions (haptoglobin [HP], heat shock protein, and fatty acid-binding protein [FABP]) were lowest in the exposed group, whereas the remaining five blood cytokine expressions (IL-2, IL-10, IFN-gamma, TNF-alpha, and TGF-beta) and two APP expressions (MT1 and MT2) were highest in the exposed group. In liver tissue, three cytokine expressions (IL-10, IFN-gamma, and TNF alpha) and two APP expressions (MT1 and MT2) were highest in the exposed group, and the remaining cytokine and APP expressions were lowest in the exposed group. Of these, the liver tissue expression of HP and FABP was significantly lowest in the exposed group (both p < 0.05). As a consequence of our findings, we suggest that a daily intake of 50 to 200 g polluted whale blubber is associated with a genotoxic decrease in HP and FABP gene expression in the liver of sledge dog and possibly of other top mammalian marine predators and consumers in the Arctic, indicating a restricted acute-phase reaction and insufficient immune response. Finally, HP and FABP liver expression appear to be new and sensitive biomarkers of organohalogen compound exposure. PMID- 17396212 TI - Biomarkers of contaminant exposure in Northern Pike (Esox lucius) from the Yukon River Basin, Alaska. AB - As part of a larger investigation, northern pike (n = 158; Esox lucius) were collected from ten sites in the Yukon River Basin (YRB), Alaska, to document biomarkers and their correlations with organochlorine pesticide (total p,p'-DDT, total chlordane, dieldrin, and toxaphene), total polychlorinated biphenyls (PCBs), and elemental contaminant (arsenic, cadmium, copper, lead, total mercury, selenium, and zinc) concentrations. A suite of biomarkers including somatic indices, hepatic 7-ethoxyresorufin O-deethylase (EROD) activity, vitellogenin concentrations, steroid hormone (17B- ustradiol and 16-kebtestosteront) concentrations, splenic macrophage aggregates (MAs), oocyte atresia, and other microscopic anomalies in various tissues were documented in YRB pike. Mean condition factor (0.50 to 0.68), hepatosomatic index (1.00% to 3.56%), and splenosomatic index (0.09% to 0.18%) were not anomalous at any site nor correlated with any contaminant concentration. Mean EROD activity (0.71 to 17.51 pmol/min/mg protein) was similar to basal activity levels previously measured in pike and was positively correlated with selenium concentrations (r = 0.88, P < 0.01). Vitellogenin concentrations in female (0.09 to 5.32 mg/mL) and male (<0.0005 to 0.097 mg/mL) pike were not correlated with any contaminant, but vitellogenin concentrations >0.01 mg/mL in male pike from multiple sites indicated exposure to estrogenic compounds. Mean steroid hormone concentrations and percent oocyte atresia were not anomalous in pike from any YRB site. Few site differences were significant for mean MA density (1.86 to 6.42 MA/mm(2)), size (812 to 1481 microm(2)), and tissue occupied (MA-%; 0.24% to 0.75%). A linear regression between MA-% and total PCBs was significant, although PCB concentrations were generally low in YRB pike (< or =63 ng/g), and MA-% values in female pike (0.24% to 0.54%) were lower than in male pike (0.32% to 0.75%) at similar PCB concentrations. Greater numbers of MAs were found as zinc concentrations increased in YRB female pike, but it is unlikely that this is a causative relationship. Histological abnormalities observed in gill, liver, spleen, and kidney tissues were not likely a result of contaminant exposure but provide information on the general health of YRB pike. The most common histologic anomalies were parasitic infestations in various organs and developing nephrons and nephrocalcinosis in posterior kidney tissues. Overall, few biomarker responses in YRB pike were correlated with chemical contaminant concentrations, and YRB pike generally appeared to be healthy with no site having multiple anomalous biomarker responses. PMID- 17396213 TI - PCBs and PCDD/FS in fish and fish products and their impact on the human body burden in Belgium. AB - The concentrations of marker PCBs (28, 52, 101, 118, 138, 153, 180) in fish have been assessed with GC-MS: an average concentration of 540 ng-PCB g(-1) fat (5.02 ng-PCB g(-1) wet weight) was observed. The average concentration of PCDD/Fs, assessed with the CALUX bioassay, amounted to 64 pg-CALUX-TEQ g(-1) fat (0.58 pg CALUX-TEQ g(-1) wet weight) and that of PCDD/Fs + dioxin-like PCBs amounted to 131 pg-CALUX-TEQ g(-1) fat (1.18 pg-CALUX-TEQ g(-1) wet weight). Results of the PCB congeners analyses show that PCB-153 is the most abundant congener in almost all samples, with also main contributions of the 138- and 180-congeners. For some species such as the sand sole and lemon sole, a fairly constant PCB content, independent of the fat percentage, was observed. For a second group of species such as whelks, cod, and whiting, a positive correlation was observed between their PCB concentration (ng g(-1) fat) and their % of fat. The relationship between marker PCBs and PCDD/Fs concentrations, when plotted on a log scale, fits a straight line (correlation coefficient r = 0.83). With our results on fish and literature data for other food products, intake of marker PCBs and PCDD/Fs could be calculated for the adult population in Belgium (19-60 years old). The Total Daily Intake (TDI) of marker PCBs (ng-PCB day(-1)) ranges between 1690 and 2210. The TDI of PCDD/Fs (pg-CALUX day(-1)) ranges between 80.5 and 122, that of PCDD/Fs + dioxin-like PCBs amounts to 151. When PCDD/Fs in fish are assessed with GC-HRMS, the TDI can be lower. The relative importance of fish regarding marker PCB intake amounts to 15-19%, while regarding PCDD/Fs intake it amounts to 34 51%. Using TDI, the body burden evolution of marker PCBs and PCDD/Fs, with age has been calculated. PMID- 17396214 TI - Rationale for, and design of, a clinical trial targeting polyamine metabolism for colon cancer chemoprevention. AB - Polyamine metabolic genes are downstream targets of several genes commonly mutated in colon adenomas and cancers. Inhibitors of ornithine decarboxylase, such as difluoromethylornithine (DFMO), and agents that stimulate polyamine acetylation and export, such as non-steroidal anti-inflammatory drugs (NSAIDS), act at least additively to arrest growth in human cell models and suppress intestinal carcinogenesis in mice. These preclinical studies provided the rationale for colon cancer prevention trials in humans. A Phase IIb clinical study comparing the combination of DFMO and the NSAID sulindac versus placebo was conducted. Endpoints were colorectal tissue polyamine and prostaglandin E2 contents and overall toxicity to participants. Participants in the Phase IIb study served as a vanguard for a randomized, placebo-controlled prospective Phase III trial of the combination of DFMO and sulindac with the primary study endpoint the prevention of colon polyps. Seventy percent of participants will have completed the three years of treatment in December 2006. PMID- 17396215 TI - Spermine and spermidine mediate protection against oxidative damage caused by hydrogen peroxide. AB - The polyamines spermidine and spermine have been hypothesized to possess different functions in the protection of DNA from reactive oxygen species. The growth and survival of mouse fibroblasts unable to synthesize spermine were compared to their normal counterparts in their native and polyamine-depleted states in response to oxidative stress. The results of these studies suggest that when present at normal or supraphysiological concentrations, either spermidine or spermine can protect cells from reactive oxygen species. However, when polyamine pools are pharmacologically manipulated to produce cells with low levels of predominately spermine or spermidine, spermine appears to be more effective. Importantly, when cells are depleted of both glutathione and endogenous polyamines, they exhibit increased sensitivity to hydrogen peroxide as compared to glutathione depletion alone, suggesting that polyamines not only play a role in protecting cells from oxidative stress but this role is distinct from that played by glutathione. PMID- 17396216 TI - Effects of creatine supplementation on glucose tolerance and insulin sensitivity in sedentary healthy males undergoing aerobic training. AB - Recent findings have indicated that creatine supplementation may affect glucose metabolism. This study aimed to examine the effects of creatine supplementation, combined with aerobic training, on glucose tolerance in sedentary healthy male. Subjects (n = 22) were randomly divided in two groups and were allocated to receive treatment with either creatine (CT) ( approximately 10 g . day over three months) or placebo (PT) (dextrose). Administration of treatments was double blind. Both groups underwent moderate aerobic training. An oral glucose tolerance test (OGTT) was performed and both fasting plasma insulin and the homeostasis model assessment (HOMA) index were assessed at the start, and after four, eight and twelve weeks. CT demonstrated significant decrease in OGTT area under the curve compared to PT (P = 0.034). There were no differences between groups or over time in fasting insulin or HOMA. The results suggest that creatine supplementation, combined with aerobic training, can improve glucose tolerance but does not affect insulin sensitivity, and may warrant further investigation with diabetic subjects. PMID- 17396217 TI - Surfaces with quenched and annealed disordered charge distributions. AB - We consider surfaces with disordered charge distribution. The disorder can be caused by mobile charges, as for example in mixed lipid bilayers, or by weakly charged surfaces where charge regulation takes place (e.g. carboxyl groups). Using Monte-Carlo simulation methods we find for quenched as well as annealed disordered charge distributions counterion densities close to the surface that are significantly larger than for ordered regularly spaced surface ions. Our field-theoretic results agree well with results obtained from Monte-Carlo simulations of the system. Furthermore, we obtain expressions for the effective interaction between charged colloids and charged rods close to a charged surface and discuss the effect of the surface-ion mobility and polarization charges on the interaction. In general, polarization effects as well as surface-ion mobility lead to a weakening of the effective interaction between charged objects. PMID- 17396218 TI - The influence of grain shape, friction and cohesion on granular compaction dynamics. AB - This article is a review of our recent and new experimental works on granular compaction. The effects of various microscopic parameters on the compaction dynamics are addressed, in particular the influence of the grain shape, the friction and the cohesion between the grains. Two dimensional and three dimensional systems are analysed. And the role of dimensionality will be emphasized. Theoretical and numerical investigations provide additional informations about that phenomenon. Indeed numerical models permit us to study the influence of some parameters not easily accessible experimentally. Our results show that the above mentioned parameters have a deep impact on the compaction dynamics. Anisotropic grains lead to two different compaction regimes separated by a "burst" of the packing fraction. Friction is observed to modify how the grains are arranged in the pile. This is confirmed by numerical simulations. Cohesive forces between particles inhibit compaction and lead to extremely low values of the packing fraction. PMID- 17396219 TI - On coupling between the orientation and the shape of a vesicle under a shear flow. AB - A simple 2D model of deformable vesicles tumbling in a shear under flow is introduced in order to account for the main qualitative features observed experimentally as shear rates are increased. The simplicity of the model allows for a full analytical tractability while retaining the essential physical ingredients. The model reveals that the main axes of the vesicle undergo oscillations which are coupled to the vesicle orientation in the flow. The model reproduces and sheds light on the main novel features reported in recent experiments [M. Mader et al., Eur. Phys. J. E. 19, 389 (2006)], namely that both coefficients A and B that enter the Keller-Skalak equation, d psi/dt = A+B cos(2 psi) (psi is the vesicle orientation angle in the shear flow), undergo a collapse upon increasing shear rate. PMID- 17396220 TI - Comment on "The properties of free polymer surfaces and their influence on the glass transition temperature of thin polystyrene films" by J.S. Sharp, J.H. Teichroeb and J.A. Forrest. AB - Sharp, Teichroeb and Forrest [J.S. Sharp, J.H. Teichroeb, J.A. Forrest, Eur. Phys. J. E 15, 473 (2004)] recently published a viscoelastic contact mechanics analysis of the embedment of gold nanospheres into a polystyrene (PS) surface. In the present comment, we investigate the viscoelastic response of the surface and conclude that the embedment experiments do not support the hypothesis of a liquid surface layer of sufficiently reduced "rheological temperature" to explain reports of very large reductions in the glass temperature of freely standing ultrathin polystyrene films. We also report some errors and discrepancies in the paper under comment that resulted in an inability to reproduce the reported calculations. We present our findings of error in a spirit of clarifying the problem of embedment of spheres into surfaces and in order that others can understand why they may not reproduce the results reported by Sharp, Teichroeb and Forrest. In the comment, we also examine the effects of the magnitude of the forces that result from the polymer surface-nanosphere particle interactions on the viscoelastic properties deduced from the embedment data and we provide a comparison of apparent surface or "rheological" temperature vs. experimental temperature that indicates further work needs to be performed to fully understand the surface embedment experiments. Finally, we comment that the nanosphere embedment measurements have potential as a powerful tool to determine surface viscoelastic properties. PMID- 17396221 TI - Phylogeny and comparative seed morphology of epiphytic and terrestrial species of Liparis (Orchidaceae) in Japan. AB - To elucidate the evolution of epiphytes in Liparis section Liparis, we examined the phylogenetic relationships of 16 species by using internal transcribed spacer regions of 18S-26S nuclear ribosomal DNA (ITS) and three chloroplast DNA regions (trnS-trnG spacer, trnL with trnL-trnF spacer, and partial matK). Results showed that the epiphytic L. fujisanensis is sister to the terrestrial L. koreana and L. kumokiri, while another epiphyte, L. truncata, is sister to the terrestrial L. krameri. Therefore, the two epiphytic species evolved from terrestrial species independently in section Liparis. Comparative seed morphology revealed that the epiphytes have larger embryos than their closely related terrestrial counterparts. A similar trend toward the increase of embryo size in the two epiphytic species belonging to closely related, but distinct clades suggests that the large embryo may have an advantage in the epiphytic lifestyle. The two epiphytic species share another character state, smaller air spaces in the seed than that of closely related terrestrial species, suggesting possible low dispersibility of the epiphytes. PMID- 17396222 TI - Physiological functions of D-amino acid oxidases: from yeast to humans. AB - D-Amino acid oxidase (DAAO) is a FAD-containing flavoenzyme that catalyzes the oxidative deamination of D-isomers of neutral and polar amino acids. This enzymatic activity has been identified in most eukaryotic organisms, the only exception being plants. In the various organisms in which it does occur, DAAO fulfills distinct physiological functions: from a catabolic role in yeast cells, which allows them to grow on D-amino acids as carbon and energy sources, to a regulatory role in the human brain, where it controls the levels of the neuromodulator D-serine. Since 1935, DAAO has been the object of an astonishing number of investigations and has become a model for the dehydrogenase-oxidase class of flavoproteins. Structural and functional studies have suggested that specific physiological functions are implemented through the use of different structural elements that control access to the active site and substrate/product exchange. Current research is attempting to delineate the regulation of DAAO functions in the contest of complex biochemical and physiological networks. PMID- 17396223 TI - Asexual reproduction: genetics and evolutionary aspects. AB - Reproduction is essential to all organisms if they are to contribute to the next generation. There are various means and ways of achieving this goal. This review focuses on the role of asexual reproduction for eukaryotic organisms and how its integration in a life cycle can influence their population genetics and evolution. An important question for evolutionary biologists as to why some organisms reproduce sexually, as opposed to asexually, is addressed. We also discuss the economic and medical importance of asexual organisms. PMID- 17396224 TI - Molecular basis for chemoprevention by sulforaphane: a comprehensive review. AB - The consumption of cruciferous vegetables has long been associated with a reduced risk in the occurrence of cancer at various sites, including the prostate, lung, breast and colon. This protective effect is attributed to isothiocyanates present in these vegetables, and sulforaphane (SF), present in broccoli, is by far the most extensively studied to uncover the mechanisms behind this chemoprotection. The major mechanism by which SF protects cells was traditionally thought to be through Nrf2-mediated induction of phase 2 detoxification enzymes that elevate cell defense against oxidative damage and promote the removal of carcinogens. However, it is becoming clear that there are multiple mechanisms activated in response to SF, including suppression of cytochrome P450 enzymes, induction of apoptotic pathways, suppression of cell cycle progression, inhibition of angiogenesis and anti-inflammatory activity. Moreover, these mechanisms seem to have some degree of interaction to synergistically afford chemoprevention. PMID- 17396225 TI - Ruminations on dietary restriction and aging. AB - Calorie restriction has been known for many decades to extend the life span of rodents. Since the more recent discovery that a long-term reduction in nutrient intake also extends life span in nearly every invertebrate model organism used for aging research, the mechanisms behind the longevity benefits of this intervention have been under intense scrutiny. While models have been developed in yeast, worms, and flies, the molecular mechanisms governing life span extension by calorie restriction remain controversial, resulting in great anticipation of mammalian studies testing these models. Here we discuss the links between nutrient reduction and enhanced longevity with emphasis on evolutionarily conserved nutrient response signaling. PMID- 17396226 TI - Investigating metal-binding in proteins by nuclear magnetic resonance. AB - Metal ions play a key role for the function of many proteins. The interaction of the metal ion with the protein and its involvement in the function of the protein vary widely. In some proteins, the metal ion is bound tightly to the ligand residues and may be the key player in the function of the protein, as in the case of blue copper proteins. In other proteins, the metal ion is bound only temporarily and loosely to the protein, as in the case of some metalloenzymes and other proteins where the metal ion acts as a cofactor necessary for the function of the protein. Such proteins are often known as metal ion-activated proteins. The review focuses on recent nuclear magnetic resonance (NMR) studies of a series of metal-dependent proteins and the characterization of the metal-binding sites. In particular, we focus on NMR techniques for studying metal binding to proteins such as chemical shift mapping, paramagnetic NMR and changes in backbone dynamics upon metal binding. PMID- 17396227 TI - Images in neuro oncology: primary extraaxial cerebellopontine angle ependymoma. AB - Posterior fossa ependymoma usually arise in the fourth ventricle. Though extension of this tumor into the cerebellopontine angle and subarachnoid space through the foramina of Luschka or Magendie is well described, a primary extraaxial cerebellopontine angle location of a posterior fossa ependymoma is distinctly uncommon. The authors report imaging in such an unusual case of a primary cerebellopontine angle ependymoma. PMID- 17396228 TI - Pathogenesis, clinical and laboratory aspects of thrombosis in cancer. AB - The relationship between increased clotting and malignancy is well recognized, though the bidirectional development of this association is often overlooked. In the challenging cancer biology, transforming genes often act in concert with numerous epigenetic factors, including hypoxia, inflammation, contact between blood and cancer cells, and emission of procoagulant vesicles from tumors, to determine a net imbalance of the hemostatic potential which is detectable by a variety of laboratory tests. Procoagulant factors, in particular, are intimately involved in all aspects of hemostatic, cell proliferation and cellular signalling systems. However, the biggest as yet unresolved question is why cancer patients develop thrombosis? Since the thrombus itself does not apparently contributes directly to the tumor biology, enhanced hemostasis activation in cancer patients may be interpreted according to the most recent biological evidences. Coagulation and cancer biology interact bidirectionally in a "vicious cycle", in which greater tumor burden supplies greater procoagulants (tissue factor, cancer procoagulant) and thrombin, which would in turn act as strong promoters of cancer growth and spread. In this perspective, thrombosis may be interpreted as a epiphenomenon of an intricate an effective biological feedback to maintain or promote cancer progression. In this review article, we briefly analyze the pathogenesis, laboratory, clinical and therapeutic features of cancer and thrombosis. PMID- 17396229 TI - Distinguishing characteristics of erythrocyte-rich and platelet-rich thrombus by intravascular ultrasound catheter system. AB - BACKGROUND: Acute coronary syndromes are associated with platelet-rich, white thrombi (WT) and erythrocyte-rich, red thrombi (RT), but their ultrasonic characteristics are not well defined. To determine whether intravascular ultrasound (IVUS) could be used to detect specific characteristics of WT and RT, two experiments were performed. METHODS: An in-vitro experiment evaluated five WT and five RT and an ex-vivo experiment evaluated thrombi from 17 atherosclerotic rabbits with disrupted plaques and overlying thrombi. Specimen were mounted flat, immersed in a saline bath and examined from the intimal surface. Thrombi were classified as WT (n = 69) or RT (n = 40) by gross inspection and histology. IVUS was performed using a 1 mm, 20 MHz transducer in a 4.8F catheter. Images were digitally converted and points integrated to account for angular and depth resolution. Sampling was performed at the water-tissue interface and four other sites at 0.3 mm radial depth increments. Signals from each depth were standardized by obtaining the ratio of each energy level to the level at the water-tissue interface. RESULTS: The average energy ratio backscattered by RT was constant with increasing tissue depth while it attenuated for WT (P < 0.005; 2 way ANOVA). RT was less homogeneous and had more backscatter compared to WT. Light and electron microscopy corroborated these observations showing WT as densely homogenous and RT with loose cellular elements. CONCLUSION: WT may be detected by its attenuated ultrasound pattern versus a non-attenuated pattern for RT by IVUS. This technique has potential for characterizing WT and RT. PMID- 17396230 TI - The thrombolytic effect of aspirin in animal model. AB - BACKGROUND: The aspirin induced platelet aggregation has been reported to be mediated through the inhibition of platelet prostaglandin synthesis. This compound has also been recently reported to stimulate nitric oxide synthesis in platelets. Since nitric oxide has been reported to produce fibrinogen/fibrinolytic effect, investigation was carried out to determine fibrinolytic effect of in vivo exposure of platelets to aspirin in normal volunteers on the fibrinolysis of the clotted platelet-rich plasma in vitro. The thrombolytic effect of aspirin in situ was also carried out by injecting aspirin solution in the mice with ADP induced formed thrombi in the coronary artery. METHODS AND RESULTS: It was found that the clotted platelet-rich plasma prepared from the volunteers (n = 10, F = 5, M = 5) who ingested 150 mg aspirin, began to undergo spontaneous and progressive fibrinolysis for 200 min at 37 degrees C with the generation of fibrin degradation products in the lysate. No such fibrinolysis could be seen in control experiments. When platelet thrombi were produced in the coronary artery of mice by injecting ADP, and these animals subsequently received intravenous injection of aspirin (4 muM final), they not only survived (P < 0.0001, n = 10) the thrombogenic assault but the lysis of the platelet thrombi was also noted in the post mortem examination. The thrombolytic effect of aspirin was found to be comparable to that of streptokinase in these animals. CONCLUSIONS: Aspirin, through the stimulation of NO synthesis, may produce thrombolysis in vivo. PMID- 17396231 TI - Reactive oxygen species plasmatic levels in ischemic stroke. AB - Oxidative stress is probably one of the mechanisms involved in neuronal damage induced by ischemia-reperfusion, and the antioxidant activity of plasma may be an important factor providing protection from neurological damage caused by stroke associated oxidative stress. The aim of this study was to investigate the status of oxidative stress, NO and ONOO(-) levels in patients with atherothrombotic and lacunar acute ischemic stroke and iNOS, eNOS and nitrotyrosine expression in the same patients. Plasma ONOO(-) levels were significantly higher in patients than in controls while NO decreases in patients in respect to controls. Densitometric analysis of bands indicated that iNOS and N-Tyr protein levels were significantly higher in patients in respect to controls. This study has highlighted a significant NO decrease in our patients compared with controls and this is most probably due to the increased expression of inducible NO synthase by the effect of thrombotic attack. In fact, the constitutive NO isoforms, which produce small amounts of NO, are beneficial, while activation of the inducible isoform of NO, which produces much more NO, causes injury, being its toxicity greatly enhanced by generation of peroxynitrite. The significant ONOO(-) increase observed in our patients, compared to controls, is most probably due to reaction of NO with O(2)(*-) . These findings suggest that free radical production and oxidative stress in ischemic stroke might have a major role in the pathogenesis of ischemic brain injury. Peroxynitrite might be the main marker of brain damage and neurological impairment in acute ischemic stroke. PMID- 17396232 TI - RNAi inhibition of Pax3/7 expression leads to markedly decreased expression of muscle determination genes. AB - Skeletal muscle regeneration by cell transplantation for the treatment of muscle diseases requires the identification and isolation of well-defined, early skeletal muscle progenitor cells. It is known that myogenesis is governed by the sequential and compound activation of the muscle determination genes, the myogenic regulatory factors (MRFs). Recently it has been proposed that the transcription factors Pax3 and Pax7 trigger the expression of the MRFs and thereby specify a novel population of cells destined to enter the myogenic program. We directly tested this hypothesis using RNA interference methodology to reduce the levels of Pax3 and Pax7 RNA in mouse embryoid bodies developing in vitro. We found that decreasing the levels of Pax3/Pax7 RNA leads to a marked and selective decrease in Myf5, MyoD and Desmin expression. Pax3 and Pax7 expressing cells from developing embryos may thus serve as the earliest known skeletal muscle progenitor cells potentially useful for cell transplantation studies. PMID- 17396233 TI - Adipocyte fatty acid-binding protein (aP2), a newly identified LXR target gene, is induced by LXR agonists in human THP-1 cells. AB - The liver X receptors (LXRalpha and LXRbeta), ligand-activated transcription factors, belong to the superfamily of nuclear hormone receptors and have been shown to play a major role in atherosclerosis by modulating cholesterol and triglyceride metabolism. In this report, we describe a novel LXR target, the adipocyte fatty acid binding protein (aP2), which plays an important role in fatty acid metabolism, adipocyte differentiation and atherosclerosis. While LXR agonists induce aP2 mRNA expression in human monocytes (THP-1 cells) and macrophages in a time- and concentration-dependent manner, they have no effect on aP2 expression in human adipocytes. The increase in aP2 mRNA level was additive when THP-1 cells were treated with LXR and PPARgamma agonists. Also, an RXR agonist induced aP2 expression in these cells. While no additive effect was observed with LXR and RXR agonists, additive effects were observed with RXR and PPARgamma agonists. GW9662, a potent PPARgamma antagonist, inhibited PPARgamma induced aP2 expression without affecting LXR-mediated aP2 expression indicating the induction is mediated directly through LXR activation. Analysis of human aP2 promoter revealed a potential LXR response element (LXRE). Gel shift data showed that the LXRalpha/RXRalpha heterodimer bound to the LXRE motif in aP2 promoter in vitro in a sequence-specific manner. Deletion and mutation analyses of the proximal aP2 promoter confirm that this is a functional LXRE. These data indicate for the first time that human macrophage aP2 promoter is a direct target for the regulation by LXR/RXR heterodimers. PMID- 17396234 TI - Resveratrol-induced mitochondrial dysfunction and apoptosis are associated with Ca2+ and mCICR-mediated MPT activation in HepG2 cells. AB - Resveratrol, a natural polyphenolic antioxidant, has been reported to possess the cancer chemopreventive potential in wide range by means of triggering tumor cells apoptosis through various pathways. It induced apoptosis through the activation of the mitochondrial pathway in some kinds of cells. In the present reports, we showed that resveratrol-induced HepG2 cell apoptosis and mitochondrial dysfunction was dependent on the induction of the mitochondrial permeability transition (MPT), because resveratrol caused the collapse of the mitochondrial membrane potential (DeltaPsi(m)) with the concomitant release of cytochrome c (Cyt.c). In addition, resveratrol induced a rapid and sustained elevation of intracellular [Ca(2+)], which compromised the mitochondrial DeltaPsi(m) and triggered the process of HepG2 cell apoptosis. In permeabilized HepG2 cells, we further demonstrated that the effect of the resveratrol was indeed synergistic with that of Ca(2+) and Ca(2+) is necessary for resveratrol-induced MPT opening. Calcium-induced calcium release from mitochondria (mCICR) played a key role in mitochondrial dysfunction and cell apoptosis: (1) mCICR inhibitor, ruthenium red (RR), prevent MPT opening and Cyt.c release; and (2) RR attenuated resveratrol induced HepG2 cell apoptotic death. Furthermore, resveratrol promotes MPT opening by lowering Ca(2+)-threshold. These data suggest modifying mCICR and Ca(2+) threshold to modulate MPT opening may be a potential target to control cell apoptosis induced by resveratrol. PMID- 17396236 TI - An improved primer extension method for detection of mRNA start-points using non radioactive digoxigenin-labeling primers. AB - An improved primer extension method using non-radioactive digoxigenin (DIG) labeling primers is described which uses a commercially-available DIG-labeling and detection system to perform alternative hybridization-based "labeling" procedure of DNA markers and DIG chemiluminescent detection assay. The time consuming annealing step and relatively low specificity of conventional protocol are also improved considerably by an application of one-step primer/mRNA annealing procedure and subsequent high-temperature reverse transcription reaction. This new protocol is convenient, simple, cost-effective and safe, and can allow the detection of even low abundance mRNA start-points in enriched poly(A)(+) RNA samples. PMID- 17396235 TI - Transcriptional regulation of intronic calcium-activated potassium channel SK2 promoters by nuclear factor-kappa B and glucocorticoids. AB - Small-conductance Ca(2+)-activated K(+) channels (SK) of the SK2 subtype are widely expressed in the central nervous system where they contribute to the control of neuronal excitability. Two SK2 isoforms, SK2-S and SK2-L, the latter representing an N-terminally extended protein of SK2-S, are expressed in similar patterns in the brain. However, our understanding of mechanisms by which the expression of SK2 is regulated is limited. We identified one functional glucocorticoid response element (GRE) at position -2248 bp and two functional nuclear factor-kappB (NF-kappaB) response elements at positions -1652 and -1586 bp in the SK2-S promoter. An increase in SK2-S promoter activity was observed in PC12 cells transiently transfected with a wild-type SK2-S promoter-luciferase reporter gene construct and treated with aldosterone or dexamethasone. The mineralocorticoid receptor (MR) antagonist spironolactone or the glucocorticoid receptor (GR) antagonist mifepristone fully inhibited aldosterone or dexamethasone activation of the SK2-S promoter, respectively. SK2-S promoter activity was also induced by the cell-permeable ceramide analog, N acetylsphingosine (C2-ceramide). Antisense oligonucleotides directed to NF-kappaB p65 or p50 suppressed SK2-S transcription induced by C2-ceramide. Deletion studies showed that only the -1586 bp NF-kappaB binding site was necessary for maximum C2-ceramide response. Finally, we showed that activation of GRs but not of MRs repressed the NF-kappaB-mediated induction of SK2-S transcription. These findings suggest a possible transcriptional cross talk between GRs and NF-kappaB in the intronic promoter regulation of SK2-S channel gene transcription. PMID- 17396237 TI - 1H, 15N and 13C resonance assignments of CG7054, a new PEBP from Drosophila melanogaster. PMID- 17396238 TI - Light-enhanced caffeic acid derivatives biosynthesis in hairy root cultures of Echinacea purpurea. AB - Light plays an important role in almost all plant developmental processes and provides the fundamental building blocks for growth, development, primary and secondary metabolism. The effects of light on growth rate and caffeic acid derivative (CADs) biosynthesis in hairy root cultures of Echinacea purpurea (Moench) were assessed. Light-grown hairy roots accumulated increased levels of anthocyanins, which became visible in outer cell layer of the cortex as a ring of purple color. The light-grown root cultures also had radially thickened morphology compared with the dark-grown controls. The growth rate and cell viability of the hairy root cultures in light did not show obvious difference in comparison with those in dark. However, biosynthesis of CADs including cichoric acid, caftaric acid, chlorogenic acid and caffeic acid was significantly increased in hairy root cultures grown in the light. The enhanced accumulation of CADs and anthocyanins in E. purpurea hairy root cultures was correlated to an observed light-stimulated activity of phenylalanine ammonium lyase (PAL). PMID- 17396239 TI - The plant cell nucleus is constantly alert and highly sensitive to repetitive local mechanical stimulations. AB - When mechanical stimulation is applied to a plant cell, the nucleus usually shows oriented movement to the site of stimulation (as a defensive response). Former researchers have revealed that applying mechanical pressure to plant tissues could line up cell division plane. A proposal, therefore, was put forward that cells inside plant tissue could receive mechanical signals from their growing neighbors to adjust their nuclear position and thus regulate the orientation of their dividing plane in order to form characteristic morphology of plant organs. To explore nuclear capacity and sensitivity to rapidly changing signals, multiple mechanical stimulations were applied to the same plant cell at intervals, either locally or at distance. The results revealed that the nucleus was highly sensitive to mechanical stimulations. It responded quickly to both local and distant stimulation by showing oriented movement toward the stimulation site. The nucleus was able to respond immediately to a second stimulation (no time lag) by starting up a second oriented movement toward the new signal; the completion of nuclear oriented movement to a first site of stimulation was not necessary for startup of a subsequent movement track to a second stimulation site, regardless of whether the second stimulation was applied ahead of or behind the moving nucleus. The nucleus responded to a second stimulation without loss of velocity, whether or not it was in a resting or moving state. This novel finding favors the proposal that growing tissues adjust the location of nuclei in cells by varying mechanical pressures; they thus control cell division according to a plan whereby organs and their constituent tissues develop in an orderly, specified manner. It appears that the enhanced sensitivity of plant cells to mechanical pressure is necessary not only in response to the external environment, but also to the developmental microenvironment inside the tissues. PMID- 17396240 TI - [Coronary stents, dual antiplatelet therapy and peri-operative problems]. AB - Up to 90% of all percutaneous coronary interventions include coronary artery stenting. Dual antiplatelet therapy, usually involving acetylsalicyl acid combined with clopidogrel, is mandatory for patients with coronary artery stents. The duration of antiplatelet therapy for bare metal stents is 3-4 weeks, for drug eluting stents 6-12 months. Preoperative discontinuation of both drugs increases the risk of stent thrombosis, continuation the risk of relevant bleeding. According to the recommendations of anaesthesiological and cardiological societies, perioperative management has to balance the risk of bleeding vs stent thrombosis. Surgery involving a high risk of bleeding can require the discontinuance of both substances. In cases of high thrombosis risk, at least the acetylsalicyl acid should be continued until the day of surgery. For patients under antiplatelet therapy scheduled for local anaesthesia, national recommendations exist. A close collaboration between the anaesthesiologist, cardiologist and surgeon is essential for appropriate pre-, intra- and postoperative management. PMID- 17396241 TI - Overactivity of the intestinal endocannabinoid system in celiac disease and in methotrexate-treated rats. AB - The endocannabinoid system is upregulated in both human inflammatory bowel diseases and experimental models of colitis. In this study, we investigated whether this upregulation is a marker also of celiac disease-induced atrophy. The levels of the cannabinoid CB(1) receptor, of the endocannabinoids, anandamide, and 2-arachidonoyl-glycerol (2-AG), and of the anti-inflammatory mediator palmitoylethanolamide (PEA) were analyzed in bioptic samples from the duodenal mucosa of celiac patients at first diagnosis assessed by the determination of antiendomysial antibodies and histological examination. Samples were analyzed during the active phase of atrophy and after remission and compared to control samples from non-celiac patients. The levels of anandamide and PEA were significantly elevated (approx. 2- and 1.8-fold, respectively) in active celiac patients and so were those of CB(1) receptors. Anandamide levels returned to normal after remission with a gluten-free diet. We also analyzed endocannabinoid and PEA levels in the jejunum of rats 2, 3, and 7 days after treatment with methotrexate, which causes inflammatory features (assessed by histopathological analyses and myeloperoxidase activity) similar to those of celiac patients. In both muscle/serosa and mucosa layers, the levels of anandamide, 2-AG, and PEA peaked 3 days after treatment and returned to basal levels at remission, 7 days after treatment. Thus, intestinal endocannabinoid levels peak with atrophy and regress with remission in both celiac patients and methotrexate-treated rats. The latter might be used as a model to study the role of the endocannabinoid system in celiac disease. PMID- 17396242 TI - Lack of association between complement factor H polymorphisms and coronary artery disease or myocardial infarction. AB - Complement factor H (CFH) plays a critical role in the protection of host cells and tissues from damage by complement activation and has been suggested to protect against the progression of atherosclerosis. A polymorphism in the CFH gene, Y402H, known to be strongly associated with age-related macular degeneration, has been analyzed in relation to coronary artery disease (CAD) in several studies with conflicting results. We investigated the association of polymorphisms of the CFH gene in two large-scale studies on CAD and myocardial infarction (MI). The AtheroGene Study included a cohort of cases with CAD (n = 1,303) prospectively followed for a median period of 6.2 years, among whom198 experienced a cardiovascular event, and a group of 483 control subjects. The AtheroGene Study population was genotyped for the Y402H, I62V, and E936D polymorphisms. There was no significant difference in genotypic or allelic frequencies between CAD cases and controls. Among cases, no significant association was found with prospective cardiovascular outcome. Many inflammatory proteins, including the C-reactive protein, were measured, and none of the polymorphisms showed an association with these markers. The Etude Cas-Temoin de l'Infarctus du Myocarde (ECTIM) Study compared 1,034 patients with MI and 1,039 controls from France and United Kingdom. The ECTIM Study population was genotyped for the Y402H polymorphism. Genotype and allele frequencies were similar in cases and controls. These results do not support an involvement of common nonsynonymous polymorphisms of the CFH gene in predisposition to CAD and its complications. PMID- 17396244 TI - [Femoral neck prostheses]. AB - The survival rates of cementless femoral neck prostheses are lower than for cementless standard stems. Nevertheless, femoral neck prostheses have the advantage of preserving the femoral neck as well as not opening the diaphyseal marrow cavity. Should the need to change the endoprosthesis arise, a standard stem anchored in the diaphysis can be implanted after resection of the femoral neck. Since there is a relationship between surgical error and the learning curve for a new implant, the surgeon should be experienced in the field of total hip arthroplasty. In addition, the implantation of femoral neck prostheses is technically demanding. Therefore, careful patient selection is necessary and patients have to be informed of the advantages as well as the disadvantages of the different implant models. PMID- 17396243 TI - Tuning cardiac performance in ischemic heart disease and failure by modulating myofilament function. AB - The cardiac myofilaments are composed of highly ordered arrays of proteins that coordinate cardiac contraction and relaxation in response to the rhythmic waves of [Ca(2+)] during the cardiac cycle. Several cardiac disease states are associated with altered myofilament protein interactions that contribute to cardiac dysfunction. During acute myocardial ischemia, the sensitivity of the myofilaments to activating Ca(2+) is drastically reduced, largely due to the effects of intracellular acidosis on the contractile machinery. Myofilament Ca(2+) sensitivity remains compromised in post-ischemic or "stunned" myocardium even after complete restoration of blood flow and intracellular pH, likely because of covalent modifications of or proteolytic injury to contractile proteins. In contrast, myofilament Ca(2+) sensitivity can be increased in chronic heart failure, owing in part to decreased phosphorylation of troponin I, the inhibitory subunit of the troponin regulatory complex. We highlight, in this paper, the central role of the myofilaments in the pathophysiology of each of these distinct disease entities, with a particular focus on the molecular switch protein troponin I. We also discuss the beneficial effects of a genetically engineered cardiac troponin I, with a histidine button substitution at C-terminal residue 164, for a variety of pathophysiologic conditions, including hypoxia, ischemia, ischemia-reperfusion and chronic heart failure. PMID- 17396245 TI - Comparative hydrolysis of O-hexyl O-2,5-dichlorophenyl phosphoramidate and paraoxon in different tissues of vertebrates. AB - The Ca(2+)-dependent and EDTA-resistant hydrolysis of O-hexyl O-2,5 dichlorophenyl phosphoramidate (HDCP) and paraoxon was studied in serum and subcellular fractions of liver, kidney and brain of hen, rat and rabbit. HDCP was the best substrate among all the tissues studied, except that of rabbit serum which showed the highest Ca(2+)-dependent paraoxon hydrolysing activity (paraoxonase). High HDCP hydrolysing activity (HDCPase) was detected in the brain tissue of the three species studied, whereas low or no paraoxonase was found. The HDCPase/paraoxonase ratio of Ca(2+)-dependent hydrolysing activities ranged from 0.5 to 83 for tissues of the same species. EDTA-resistant HDCPase activity was more than 50% of the total activities in hen tissues, with an almost undetectable Ca(2+)-dependent paraoxonase activity in most organs. The same response was observed in rat tissues, except for serum where the Ca(2+)-dependent HDCPase and paraoxonase activities were higher (70 and 25% of total activities, respectively). EDTA-resistant HDCPase and paraoxonase activities represented less than 25% of all activities in rabbit tissues. Paraoxon has traditionally been the substrate for measuring organophosphorus hydrolysing activities. However, HDCP could be a good substrate in addition to paraoxon for monitoring other phosphotriesterases in biological tissues. PMID- 17396247 TI - Determination of lead in blood by laser ablation ICP-TOF-MS analysis of blood spotted and dried on filter paper: a feasibility study. AB - Screening programs for pediatric lead (Pb) poisoning routinely collect blood by finger stick onto specially designed filter paper (FP) for subsequent laboratory testing. Here, laser ablation inductively coupled plasma time-of-flight mass spectrometry is evaluated as a novel method for the determination of Pb in blood spotted onto FP cards. The method requires no sample pretreatment and may be particularly suitable for rapidly screening a large number of samples. Signal intensity from the ICP-MS strongly correlated (r (2) = 0.996) with Pb target values for five blood cards from a laboratory proficiency testing program. Better precision was obtained by replicate ablation line scans (<15% rsd) compared to a grid of 25 individual ablation spots (>30% rsd). Because <2% of the sample is consumed in the laser ablation analysis, selected samples can also be analyzed by traditional techniques or archived. Lead isotope ratios determined during the same total-Pb analysis appear to be effective for discerning contamination extraneous to the blood sample. Determining that an elevated result was due to contamination may negate the need for retesting with invasive pediatric venipuncture. Some elements monitored along with Pb exhibited signal profiles conducive to potential quantitation (Ca, V, Fe, Cu Zn), whereas others did not (Cr, Mn, Co, Ni, Cd, Hg, Tl, U). Copper, Ca, and V were inversely correlated with Pb (r = -0.89, -0.90, and -0.92, respectively). Scanning electron microscope images revealed surface modality differences between FP spotted with fresh whole blood and reconstituted freeze-dried blood reference material. PMID- 17396246 TI - Gene-environment interactions in vulnerability to cocaine intravenous self administration: a brief social experience affects intake in DBA/2J but not in C57BL/6J mice. AB - RATIONALE: Individual differences in cocaine-taking behavior and liability to develop abuse are clearly observed, but underlying mechanisms are still poorly understood. A role for gene-environment interactions has been proposed but remains hypothetical. OBJECTIVES: We investigated whether gene-environment interactions influence intravenous cocaine self-administration (SA) in mice. We tested the effect of a past short group housing experience on cocaine SA in two inbred strains of mice, the C57BL/6J (C57) and DBA/2J (DBA). METHODS: Adult C57 and DBA mice were individually housed upon arrival in the laboratory. After 3 weeks, half of the animals of each strain were group housed for 19 days. One week after the end of group housing, cocaine SA or measurement of brain cocaine levels took place. RESULTS: Individually and ex-group-housed C57 mice did not differ for cocaine SA. On the contrary, the ex-group-housed DBA mice showed an upward shift in the dose-response curve as compared to individually housed DBA. Differences in brain cocaine levels could not account for the observed behavioral differences. CONCLUSIONS: These results demonstrate that vulnerability to cocaine reinforcing effects can be affected by gene-environment interactions. We propose a mouse model for the characterization of gene-environment interactions in the vulnerability to cocaine-taking behavior. PMID- 17396248 TI - The natural fingerprint of stable isotopes--use of IRMS to test food authenticity. PMID- 17396249 TI - Cue and reward signals carried by monkey entorhinal cortex neurons during reward schedules. AB - Ablation of entorhinal/perirhinal cortices prevents learning associations between visual stimuli used as cues in reward schedules and the schedule state. Single neurons in perirhinal cortex are sensitive to associations between the cues and the reward schedules. To investigate whether neurons in the entorhinal cortex have similar sensitivities, we recorded single neuronal activity from two rhesus monkeys while the monkeys performed a visually cued reward schedule task. When the cue was related to the reward schedules, the monkeys made progressively fewer errors as the schedule state became closer to the reward state, showing that the monkeys were sensitive to the cue and the schedule state. Of 75 neurons recorded in the entorhinal cortex during task performance, about 30% responded. About half of these responded after cue presentation. When the relation of the cue to the reward schedules was random, the cue-related responses disappeared or lost their selectivity for schedule states. The responses of the entorhinal cortex neurons are similar to responses of perirhinal cortex neurons in that they are selective for the associative relationships between cues and reward schedules. However, they are particularly selective for the first trial of a new schedule, in contrast to perirhinal cortex where responsivity to all schedule states is seen. A different subpopulation of entorhinal neurons responded to the reward, unlike perirhinal neurons which respond solely to the cue. These results indicate that the entorhinal signals carry associative relationships between the visual cues and reward schedules, and between rewards and reward schedules that are not simply derived from perirhinal cortex by feed-forward serial processing. PMID- 17396250 TI - Acute renal failure due to bilateral uric acid lithiasis in infants. AB - Acute renal failure (ARF) is one of the complications of urolithiasis, but the role of medical treatment to relieve urinary obstruction in children with ARF is uncertain. We report on infants with acute obstructive uric acid lithiasis. We describe presentation features as well as diagnosis methods and medical treatment in five infants who were admitted to our institution with ARF due to uric acid lithiasis. The medical treatments for all patients were fluid liberalization, urine alkalinization, and oral allopurinol. Two children underwent urinary diversion. Within 8 h, urine output improved in all patients, and the stones passed spontaneously. All obstructed kidneys were relieved with medical treatment, and no renal sequel remained. So this series has showed a role of medical therapy in acute obstructive uric acid lithiasis. PMID- 17396251 TI - Efficacy of expulsive therapy using nifedipine or tamsulosin, both associated with ketoprofene, after shock wave lithotripsy of ureteral stones. AB - Extracorporeal shock wave lithotripsy (ESWL) is currently considered one of the main treatments for ureteral stones. Some studies have reported the effectiveness of pharmacologic therapies (calcium antagonists or alpha-blockers) in facilitating ureteral stone expulsion after ESWL. We prospectively evaluated the efficacy, after ESWL, of nifedipine on upper-middle ureteral stones, and tamsulosin on lower ureteral stones, both associated to ketoprofene as anti-edema agent. From January 2003 to March 2005 we prospectively evaluated 113 patients affected by radiopaque or radiolucent ureteral stones. Average stone size was 10.16 +/- 2.00 mm (range 6-14 mm). Thirty-seven stones were located in the upper ureter, 27 in the middle ureter, and 49 in the lower ureter. All patients received a single session of ESWL (mean number of shock waves: 3,500) by means of a Dornier Lithotripter S (mean energy power for each treatment: 84%). Both ultrasound and X-ray were used for stone scanning. After treatment, 63 of 113 patients were submitted to medical therapy to aid stone expulsion: nifedipine 30 mg/day for 14 days administered to 35 patients with upper-middle ureteral stones (group A1) and tamsulosin 0.4 mg/day for 14 days administered to 28 patients with stones located in the distal ureter (group A2). The remaining 50 patients were used as a control group (29 upper-middle ureteral stones-B1-and 21 lower ureteral stones-B2-), receiving only pain-relieving therapy. No significant difference in stone size between the groups defined was observed. Stone clearance was assessed 1 and 2 months after ESWL by means of KUB, ultrasound scan and/or excretory urography. A stone-free condition was defined as complete stone clearance or the presence of residual fragments smaller than 3 mm in diameter. The stone-free rates in the expulsive medical therapy group were 85.7 and 82.1% for the nifedipine (A1) and tamsulosin (A2) groups respectively; stone-free rates in the control groups were 51.7 and 57.1% (B1 and B2, respectively). Five patients (14.3%) in group A1, 5 (17.8%) in group A2, 14 (48.3%) in group B1 and 9 (42.8%) in group B2 were not stone-free after a single ESWL session and required ESWL re treatment or an endoscopic treatment. Medical therapy following ESWL to facilitate ureteral stone expulsion results in increased 1- and 2-month stone free rates and in a lower percentage of those needing re-treatment. The efficacy of nifedipine for the upper-mid ureteral tract associated with ketoprofene makes expulsive medical therapy suitable for improving overall outcomes of ESWL treatment for ureteral stones. PMID- 17396252 TI - Interleukin-18 gene polymorphism, but not interleukin-2 gene polymorphism, is associated with rheumatoid arthritis. AB - To investigate whether polymorphisms of IL-2 and IL-18 genes are associated with rheumatoid arthritis (RA), polymorphisms of IL-2 and IL-18 genes were detected by polymerase-chain-reaction-based restriction analysis in the patients with RA and normal controls. The results for the IL-18 gene revealed a significant difference between the patients and the normal controls (p = 0.000003), but there was no significant difference for the IL-2 gene (p = 0.876). The IL-18 gene 105A allele was associated with RA in Chinese patients. Individuals possessing the 105A allele had a higher incidence of RA. A lack of association of IL-2 gene polymorphism between RA patients and healthy individuals was noted. The results of this study provide genetic evidence that IL-18-105A/C polymorphism may play an effective role in RA. PMID- 17396253 TI - Metagenomics, biotechnology with non-culturable microbes. AB - Metagenomics as a new field of research has been developed over the past decade to elucidate the genomes of the non-cultured microbes with the goal to better understand global microbial ecology on the one side, and on the other side it has been driven by the increasing biotechnological demands for novel enzymes and biomolecules. Since it is well accepted that the majority of all microbes has not yet been cultured, the not-yet-cultivated microbes represent a shear unlimited and intriguing resource for the development of novel genes, enzymes and chemical compounds for use in biotechnology. However, with respect to biotechnology, metagenomics faces now two major challenges. Firstly, it has to identify truly novel biocatalysts to fulfil the needs of industrial processes and green chemistry. Secondly, the already available genes and enzymes need to be implemented in production processes to further prove the value of metagenome derived sequences. PMID- 17396254 TI - Genetic organization of the biosynthetic gene cluster for the indolocarbazole K 252a in Nonomuraea longicatena JCM 11136. AB - Indolocarbazole metabolite K-252a is a natural product that was previously reported as a potent protein kinase C inhibitor with in vitro and in vivo potency. From a biosynthetic viewpoint, this compound possesses structurally interesting features such as an unusual furanosyl sugar moiety, which are absent in the well-studied staurosporine and rebeccamycin. A cosmid library from genomic DNA of Nonomuraea longicatena JCM 11136 was constructed and screened for the presence of genes to be involved in the biosynthesis of indolocarbazole K-252a. Using as a probe an internal fragment of vioB, a Chromobacterium violaceum gene encoding a multifunctional enzyme that catalyzes tryptophan decarboxylation and condensation reaction in violacein biosynthesis, we isolated a DNA region that directed the biosynthesis of K-252a when introduced into the heterologous expression host Streptomyces albus. Sequence analysis of 45 kb revealed genes for indolocarbazole core formation, glycosylation, and sugar methylation, as well as a regulatory gene and two resistance/secretion genes. The cloned genes should help to elucidate the molecular basis for indolocarbazole biosynthesis and generate new indolocarbazole analogues by genetic engineering. PMID- 17396255 TI - Characterization of estrogen-degrading bacteria isolated from an artificial sandy aquifer with ultrafiltered secondary effluent as the medium. AB - This study investigated the aerobic and anoxic biodegradation of four estrogens [estrone (E1), estradiol (E2), estriol (E3), and the synthetic 17 alpha ethinylestradiol (EE2)] in microcosms constructed with marine sand and ulftrafiltered (UF) secondary effluent. Three estrogen-degrading bacteria, LHJ1, LHJ3, and CYH, were isolated. Based on gram-stain morphology and 16S rRNA sequence homology, LHJ1 and LHJ3 belong to the genus Acinetobacter and Agromyces, respectively; CYH matched to 95% with the genus Sphingomonas. Aerobically LHJ3 degrades E3, CYH degrades E1, and all three isolates oxidize E2 to E1. Under anoxic conditions, CYH degrades E1 and LHJ3 degrades E2, whereas E3 and EE2 were not degraded by the three isolates; EE2 was transformed in microcosms incubated with site ground water. The degradation kinetics of E1 and E2 by CYH and E2 by LHJ3 under aerobic conditions was linearly correlated with the initial concentration, which ranged from 50 to 2,000 microg/l. The degradation of E1 by CYH under anoxic conditions followed Michaelis-Menten kinetics. 16 alpha Hydroxyestrone was found to be a transient transformation product of E3 under aerobic conditions. PMID- 17396256 TI - Aortic embolism of cement: a rare complication of lumbar percutaneous vertebroplasty. AB - An aortic embolus of cement occurred in a 72-year-old woman after lumbar percutaneous vertebroplasty for breast cancer metastasis. A CT scan during the procedure revealed cement migration into the aorta via the lumbar artery. The cement embolus in the aorta, along with the hook-shaped cement fragment in the lumbar artery, was asymptomatic. Thereafter, a CT scan 4 months later still showed the presence of cement in the aorta and the lumbar artery at the level of the vertebroplasty. The patient is free of complaints after 4 months without anticoagulant treatment. This rare complication occurred because the collateral arterial supplies to the breast metastasis and the retrograde migration of cement during the vertebroplasty were not recognized. Proper techniques can minimize the risk of arterial embolism caused by cement during the procedure. PMID- 17396257 TI - The emergence of MRI and MSCT cardiac imaging: nuclear cardiology is not the only actor on the stage. PMID- 17396258 TI - EANM procedure guideline for 32P phosphate treatment of myeloproliferative diseases. AB - INTRODUCTION: (32)P phosphate was the first therapeutic radioisotope, used in leukaemia about 70 years ago. Since then, many new agents for haematological proliferations have been introduced successfully. Today there remains a distinct subgroup of elderly patients with polycythaemia vera and essential thrombocythaemia for whom (32)P is the most optimal treatment option, an assertion supported by two large studies with long follow-up. PURPOSE: The purpose of this guideline is to assist the nuclear medicine physician in treating and managing patients who may be candidates for (32)P phosphate therapy. PMID- 17396259 TI - Attitudes to blood transfusion post arthroplasty surgery in the United Kingdom: a national survey. AB - Five hundred orthopaedic surgeons and 336 anaesthetists were surveyed to assess current UK attitudes towards transfusion practice following arthroplasty surgery. Seventy-two percent of surgeons and 73% of anaesthetists responded to the survey. In an uncomplicated patient following total hip arthroplasty, 53.2% of surgeons and 63.1% of anaesthetists would transfuse at or below a haemoglobin (Hb) level of 8 g/dL. Surgeons tended to be more aggressive in their attitudes, with a mean transfusion threshold of 8.3 g/dL compared to 7.9 g/dL for anaesthetists (p<0.01), and with 97% of surgeons transfusing two or more units compared to 78% of anaesthetists (p<0.01). This threshold Hb increased if the patient was symptomatic (surgeons 9.3 g/dL, anaesthetists 8.8 g/dL, p<0.05) or was known to have pre-existing ischaemic heart disease (surgeons 9.0 g/dL, anaesthetists 9.2 g/dL, p<0.05). A wide variability in attitudes and practices is demonstrated, and the development and adoption of consensus guidelines needs to be encouraged if efforts to reduce the use of blood products are to succeed. PMID- 17396260 TI - Avascular necrosis of the femoral head: inter- and intraobserver variations of Ficat and ARCO classifications. AB - Diagnosis of avascular necrosis (AVN) of the femoral head depends on the combination of clinical symptoms and evaluation of radiographs and/or magnetic resonance imaging (MRI). To evaluate the evolution of AVN, the Ficat and the Association Research Circulation Osseous (ARCO) classification are commonly used to assess both imaging modalities. For comparison reasons, these classifications need to be reliable and reproducible to provide sufficient therapy options for the patient. Therefore, the aim of our study was to evaluate the interobserver reliability and the intraobserver reproducibility of these classifications. Patients with suspected AVN were examined using either radiographs or radiographs and MRI. The radiographs and/or MR images were reviewed initially and at 3 months by two general orthopaedic surgeons, two orthopaedic residents, and two general radiologists using the Ficat classification for radiographs and MR images as well as the ARCO classification for MR images only. In all, 38 patients (54 hips) were enrolled. There were 10 patients who presented with radiographs and 28 patients with radiographs and MR scans. Paired comparisons revealed a mean interobserver kappa reliability coefficient of 0.39 for the first and of 0.32 for the second review using the Ficat classification for radiographs, whereas for the MR images a mean of 0.39 in the first and of 0.34 in the second reading resulted. The MRI evaluation using the ARCO classification resulted in a mean interobserver reliability coefficient of 0.37 in the first and of 0.31 in the second reading. The mean kappa value for intraobserver reproducibility using the Ficat classification was 0.52 for radiographs and 0.50 for MR images, whereas a reproducibility of 0.43 resulted for the ARCO classification. This study showed poor interobserver reliability and fair intraobserver variability, diminishing any meaningful comparison of studies using the Ficat as well as the ARCO classification. Thus, the Ficat and ARCO staging systems are still not sufficient to reliably assess the status of AVN alone. PMID- 17396261 TI - Highly aggressive ALK-positive anaplastic large cell lymphoma with a leukemic phase and multi-organ involvement: a report of three cases and a review of the literature. AB - Anaplastic large cell lymphoma (ALCL) is an aggressive neoplasm of T- or null cell phenotype and is recognized as a distinct clinicopathologic subtype of non Hodgkin lymphoma (NHL) in the revised World Health Organization (WHO) classification of hematopoietic neoplasms. It is rarely associated with leukemic phase. Most cases with leukemic involvement are the small cell variant of ALCL. These cases often lack the pleomorphism seen in the common variant of ALCL and may be misdiagnosed. We report a series of three patients who presented with leukemic phase ALCL. The patients included an 11-year-old boy, a 29-year-old man, and a 59-year-old woman. The clinical and pathologic features of these cases are reviewed. The patients in our case series with leukemic phase ALCL exhibited rare clinical features. The patients presented with massive extranodal disease involving cerebrospinal fluid (CSF), liver, spleen, lungs, and bone marrow. CSF involvement was documented morphologically as well as by flow cytometry in two patients. Two of the patients had small cell variant and the third patient had common type ALCL. The neoplastic cells in all three patients were ALK positive; however these patients died within months of diagnosis. Leukemic phase ALCL is rare, and behaves in an aggressive manner. Some, but not all, cases in the literature presenting with peripheral blood involvement had small cell variant ALCL, as seen in two of our cases. The leukemic phase of ALCL should be considered when a T-cell leukemia with unusual morphologic features is encountered. PMID- 17396262 TI - A novel Bcl-2 small molecule inhibitor 4-(3-methoxy-phenylsulfannyl)-7-nitro benzofurazan-3-oxide (MNB)-induced apoptosis in leukemia cells. AB - A novel small molecule inhibitor, 4-(3-methoxy-phenylsulfannyl)-7-nitro benzofurazan-3-oxide (MNB), competes with the Bak BH3 peptide to bind Bcl-2 protein with a binding affinity of IC(50) = 0.70 microM, as assessed by a fluorescence polarization based binding assay. HL-60 cells express the highest levels of Bcl-2 among the cell lines examined. Treated with 5 microM of MNB only for 6 h, 85% of HL-60 cells were detected to undergo apoptosis. Pan-caspase inhibitor, Z-VAD-FMK, blocks MNB-induced apoptosis in HL-60 cells. Caspase-2, caspase-3, caspase-7, caspase-8, caspase-9, and PARP activation were observed at as early as 4 to 6 h of MNB treatment. In addition, it has been confirmed that the caspase-3 specific inhibitor, Z-DEVD-FMK, blocks the activation of caspase-8 in MNB-treated HL-60 cells. MNB treatment does not change Bcl-2 or Bax expression level in HL-60 cells, but causes Bid cleavage. Further experiments have illustrated that MNB inhibits the heterodimerization of Bcl-2 with Bax or Bid, reduces the mitochondrial membrane potential (DeltaPsimt), and induces cytochrome c release from mitochondria in HL-60 cells. These results suggest that MNB induces apoptosis in HL-60 by inhibiting the heterodimerization of Bcl-2 with pro apoptosis Bcl-2 members, resulting in a decrease in the mitochondrial membrane potential and cytochrome c release, activation of caspases and PARP; it is a caspase-dependent process in which the activation of caspase-8 is dependent on the mitochondrial apoptosis signal transduction pathway. MNB prolongs the life spans of HL-60 bearing mice, potently kills fresh AML and ALL cells, indicating that it has the potential to be developed to treat leukemia. PMID- 17396263 TI - Cell death in response to antimetabolites directed at thymidylate synthase. AB - PURPOSE: Thymidylate synthase (TS) is an indispensable enzyme in the de novo biosynthesis of TMP during DNA replication and cell growth, and has, therefore, been an important target for several classes of antimetabolites used in cancer chemotherapy. While most investigations of the action of TS-directed agents have focused on apoptosis as the primary means of cell death, little is known regarding the role, if any, of non-apoptotic mechanisms. In the present study, we have examined the mode of cell death induced by several TS inhibitors. METHODS: Apoptosis and necrosis in response to TS inhibitors was assessed. The roles of caspases and the transcriptional regulator nuclear factor kappa B (NFkappaB) in drug-induced cell death were analyzed. Finally, drug-mediated changes in expression of several proteins involved in regulation of apoptosis were analyzed. RESULTS: Though human colon tumor cells exposed to TS inhibitors undergo classical apoptosis, it is not the predominant mechanism of response; rather, a necrosis-like mechanism prevails. The apoptotic response to TS inhibitors is caspase-dependent, and is promoted by NFkappaB. In contrast, the necrosis-like response is independent of both caspases and NFkappaB. Exposure to TS inhibitors induces PARP cleavage, but does not alter expression of the pro or activated forms of caspases-3 or caspases-8, Fas, or FasL. Treatment with the death inducing cytokine TNFalpha, like TS inhibitors, results in a limited extent of apoptosis that is both caspase- and NFkappaB-dependent; however, unlike TS inhibitors, the cytokine does not induce necrosis. CONCLUSION: Classical apoptosis occurs to a limited extent in human colon tumor cells exposed to TS inhibitors, with caspase-independent necrosis being the prinicipal mechanism of cell death. We suggest that the role of necrosis and necrosis-like mechanisms should be considered in future studies of the action of TS-directed antimetabolites, as well as other chemotherapeutic agents. PMID- 17396264 TI - Hydroxyl radical scavenger ameliorates cisplatin-induced nephrotoxicity by preventing oxidative stress, redox state unbalance, impairment of energetic metabolism and apoptosis in rat kidney mitochondria. AB - Nephrotoxicity is the major dose-limiting factor of cisplatin chemotherapy. Reactive oxygen species generated in mitochondria are thought to be the main cause of cellular damage in such injury. The present study examined, in vivo, the protective potential of the hydroxyl radical scavenger dimethylthiourea (DMTU) against cisplatin-induced effects on renal mitochondrial bioenergetics, redox state and oxidative stress. Adult male Wistar rats (200 to 220 g) were divided into four groups of eight animals each. The control group was treated only with an intraperitoneal (i.p.) injection of saline solution (1 ml/100 g body weight). The second group was given only DMTU (500 mg/kg body weight, i.p, followed by 125 mg/Kg, i.p., twice a day until they were killed). The third group was given a single injection of cisplatin (10 mg/kg body weight, i.p.). The fourth group was given DMTU (500 mg/kg body weight, i.p.), just before the cisplatin injection (10 mg/kg body weight, i.p.), followed by injections of DMTU (125 mg/kg body weight, i.p.) twice a day until they were killed. Animals were killed 72 h after the treatment. Besides not presenting any direct effect on mitochondria, DMTU substantially inhibited cisplatin-induced mitochondrial injury and cellular death by apoptosis, suppressing the occurrence of acute renal failure. All the following cisplatin-induced effects were prevented by DMTU: (1) increased plasmatic levels of creatinine and blood urea nitrogen (BUN); (2) decreased ATP content, calcium uptake and electrochemical potential; (3) oxidation of lipids, including cardiolipin; and oxidation of proteins, including sulfhydryl, and aconitase enzyme, as well as accumulation of carbonyl proteins; (4) depletion of the antioxidant defense (NADPH and GSH) and (5) increased activity of the apoptosis executioner caspase-3. Our findings show the important role played by mitochondria and hydroxyl radicals in cisplatin-induced nephrotoxicity, as well as the effectiveness of DMTU in preventing the renal mitochondrial damage caused by cisplatin. These results strongly suggest that protection of mitochondria by hydroxyl radical scavengers may be an interesting approach to prevent the kidney tissue damage caused by cisplatin-chemotherapy. PMID- 17396265 TI - Pharmacokinetics of oxaliplatin in patients with severe hepatic dysfunction. AB - PURPOSE: Data are lacking on the pharmacokinetics of oxaliplatin in patients with severe hepatic dysfunction. The aim of this study was to determine the pharmacokinetic parameters of platinum after administration of oxaliplatin in cancer patients with severe hepatic impairment due to extended metastases into the liver. PATIENTS AND METHODS: Two female breast cancer patients and one male colon cancer patient were treated with oxaliplatin monotherapy at 130 mg/m(2) given as a 3-h intravenous infusion. The patients exhibited bilirubin concentrations of 9.6, 22.5 and 41.1 mg/dl indicating severe hepatic dysfunction. Serial blood samples were collected immediately before treatment, and at fixed intervals up to 27 h after start of therapy. Platinum concentrations in plasma, ultrafilterable plasma, and whole blood were determined using a validated flameless atomic absorption spectrometry (FAAS) method. Pharmacokinetic data analysis was performed assuming a two-compartment model. Individual pharmacokinetic parameters were compared with a reference population with normal hepatic function. RESULTS: The area under the curve (AUC from 0 to infinity) as well as the elimination half-life of platinum in ultrafilterable plasma were substantially increased and clearance accordingly decreased in the three patients with severe hepatic dysfunction. In plasma and whole blood, the deviations from the reference population were less pronounced. However, partial AUC from 0 up to 2 h after end of infusion reflecting better the exposure with cytotoxic platinum species was not different or only slightly altered. Moreover, no acute oxaliplatin-associated neurotoxicity was observed. CONCLUSIONS: The comparable platinum exposure early after administration in conjunction with the lack of acute toxicity do not support a dose reduction of oxaliplatin in patients with markedly elevated bilirubin concentrations. However, a larger number of patients must be examined before valid dose recommendations can be derived. PMID- 17396266 TI - A phase II trial of gemcitabine plus capecitabine for patients with advanced pancreatic adenocarcinoma. AB - PURPOSE: While gemcitabine (GEM) is widely accepted for the treatment of advanced pancreatic cancer, capecitabine (CAP) has shown single agent activity and promising efficacy in combination with GEM. This phase II study was conducted to evaluate the efficacy and toxicity of GEM combined with dose escalated 14-day CAP as first-line chemotherapy for advanced pancreatic cancer. In addition, we also analyzed the correlation between CA19-9 response and clinical outcomes. METHODS: Patients had advanced pancreatic adenocarcinoma, no prior systemic chemotherapy other than that given concurrently with radiation therapy, at lease one measurable disease, and adequate organ functions. The patients were treated with GEM 1,000 mg/m2 IV on days 1, 8 and CAP 1,000 mg/m2 twice a day PO on days 1-14, in 21-day cycles. RESULTS: The objective RR among 45 patients was 40.0% (95% CI; 25.1-54.9), including 1CR (2.2%). The median TTP and OS were 5.4 months (95% CI; 1.8-9.0) and 10.4 months (95% CI; 6.2-14.5), respectively. Patients with >or=25% decline of serum CA19-9 had significantly better outcomes in terms of TTP and OS than those who did not (P < 0.03). The most frequent, grade 3-4, non-hematologic toxicity was hand-foot syndrome (6.7%). CONCLUSIONS: The combination of GEM with dose escalated 14-day CAP is well tolerated and offers encouraging activity in the treatment of advanced pancreatic cancer. In addition, CA19-9 response correlates well with clinical outcomes in this population. PMID- 17396267 TI - Approximating genealogies for partially linked neutral loci under a selective sweep. AB - Consider a genetic locus carrying a strongly beneficial allele which has recently fixed in a large population. As strongly beneficial alleles fix quickly, sequence diversity at partially linked neutral loci is reduced. This phenomenon is known as a selective sweep. The fixation of the beneficial allele not only affects sequence diversity at single neutral loci but also the joint allele distribution of several partially linked neutral loci. This distribution can be studied using the ancestral recombination graph for samples of partially linked neutral loci during the selective sweep. To approximate this graph, we extend recent work by Etheridge et al. (Ann Appl Probab 16:685-729, 2006) and Schweinsberg and Durrett (Ann Appl Probab 15:1591-1651, 2005) using a marked Yule tree for the genealogy at a single neutral locus linked to a strongly beneficial one. We focus on joint genealogies at two partially linked neutral loci in the case of large selection coefficients alpha and recombination rates rho = theta(alpha/log alpha) between loci. Our approach leads to a full description of the genealogy with accuracy of theta((log alpha)(-2)) in probability. As an application, we derive the expectation of Lewontin's D as a measure for non-random association of alleles. PMID- 17396268 TI - [Teratoma of the ovary. Clinical and pathological differences between mature and immature teratomas]. AB - Teratomas are the most frequent germ cell tumors of the ovary. Two main groups can be distinguished: mature and immature teratomas. Mature teratomas are benign tumors, which are most often composed of derivatives of two or three germ cell layers. Only in rare cases is the transition into a malignant tumor observed (most often squamous cell carcinoma). In contrast, immature teratomas are malignant ovarian tumors. They contain immature tissue elements in addition to the mature components, most often consisting of immature neural tissue. Histologically, this tumor component can be identified as neurotubules or rosettes. The proportion of immature tissue elements defines the grade of immaturity. Four grades have been defined in to the WHO classification. Grade 0 represents a mature teratoma. With the exception of childhood cases, grade 2 and 3 immature teratomas are treated with chemotherapy. In childhood cases, foci of yolk sac tumor (YST) must be looked for, since this determines the prognosis. If a focus of YST is present, the patient is treated with chemotherapy. Both in cases of mature and immature teratoma, peritoneal implants can be found (gliomatosis peritonei), which are also graded. In cases of immature peritoneal implants, patients are also treated with chemotherapy. Gliomatosis peritonei is most likely derived from metaplasia of subperitoneal stem cells; it does not represent a metastatic disease of the ovarian teratoma. PMID- 17396269 TI - [Therapeutic standards for ovarian cancer]. AB - Epithelial ovarian cancer is the gynecological tumor with the highest mortality rate. Recently, numerous findings have emerged on the status and extent of surgical therapy for this disease. Innovations in the chemotherapy of primary and recurrent disease are also plentiful. It is essential for the improvement of the patients' situation to transfer these new developments, already integrated into national and international standards and guidelines, into clinical practice. PMID- 17396271 TI - Giving aged care a voice. PMID- 17396270 TI - Subepyndemal hemangioblastomas of the cervicomedullary junction: lessons learned in the management of two cases. AB - OBJECTIVE: This retrospective case series analyzes two cases of hemangioblastomas in the cervicomedullary junction. METHODS: A survey of the pediatric staff and of the operative reports from medical records with a review of the literature and medical records of patients with the condition was conducted. RESULTS: Two patients were successfully treated surgically. CONCLUSION: Surgery is the treatment of choice for hemangioblastomas of the cervicomedullary junction. Careful monitoring for unique complications, treatment of the tumor as a vascular malformation, and screening for von Hippel Lindau must all be employed to safely care for this challenging group of patients. PMID- 17396272 TI - OH: Wrongful termination suit on 'public policy': failure to comply with 180 day limitations fatal. PMID- 17396273 TI - By the way, doctor. I heard that taking a proton-pump inhibitor could cause hip fractures. I've been taking 20 mg of Prilosec every day for a year. Should I be concerned? PMID- 17396275 TI - [Fewer cancer deaths in the USA]. PMID- 17396274 TI - Leishmaniasis. AB - Epidemiology, disease patterns, immunology, diagnosis, treatment and control measures of leishmaniasis are described. Various issues relating to leishmaniasis are highlighted: the relative lack of importance given to this disease compared with other infections, climate change and its possible impact on extension of endemicity of this infection, and new diagnostic tests which are improving diagnosis, especially in resource poor areas. Other important aspects discussed include the potential for newer oral therapy to change the way this disease is managed; Leishmania-HIV coinfection and groups at risk; and development of an effective vaccine. PMID- 17396276 TI - Retroperitoneal fibrosis after chemotherapy: part 2. PMID- 17396277 TI - The motion: open partial nephrectomy is the standard of care for small resectable solid renal masses. PMID- 17396278 TI - Against the motion: open partial nephrectomy is the standard of care for small resectable solid renal masses. PMID- 17396279 TI - Re: twenty years of experience with an ileal orthopedic low pressure bladder substitute--lessons to be learned. PMID- 17396280 TI - [Ultrasound 2006. Abstracts of the 30th Three-country meeting of the German, Austrian and Swiss Societies of Ultrasound in Medicine, 18-21 October 2006, Graz]. PMID- 17396282 TI - Abstracts of WorldMEN 2006 Workshop, Marseille, France, September 7-10, 2006. PMID- 17396281 TI - Abdominal sacrocolpopexy with Burch colposuspension to reduce urinary stress incontinence. PMID- 17396283 TI - The use of octyl-2-cyanoacrylate (Dermabond) tissue adhesive for skin closure in head and neck surgery. PMID- 17396284 TI - Balancing drug availability and patient safety. PMID- 17396285 TI - Psychiatric drug discovery on the couch. AB - The past two decades have been a barren time for the discovery of novel drugs for psychiatric disorders, in particular those that could revolutionize disease treatment. In our first 'Viewpoint' article this month, we present the opinions of experts from across the field of neuroscience on how the effectiveness of psychiatric drug discovery could be enhanced. PMID- 17396286 TI - Big pharma slims down to bolster productivity. PMID- 17396287 TI - Cancer nanotechnology: small, but heading for the big time. PMID- 17396288 TI - An audience with... Paul Herrling. Interview by Joanna Owens. PMID- 17396289 TI - Multiple myeloma therapies. PMID- 17396290 TI - Posaconazole. PMID- 17396291 TI - Recurrence of hepatocellular carcinoma: it's in the genes? PMID- 17396292 TI - Are we reporting the same thing? PMID- 17396293 TI - The evolution of caval implantation in liver transplantation. PMID- 17396294 TI - Incidence of initial local therapy among men with lower-risk prostate cancer in the United States. PMID- 17396295 TI - [Abstracts of the Berlin-Brandenburg Society of Ophthalmology, 1-2 December 2006, Berlin-Steglitz]. PMID- 17396296 TI - A. Richard Green. PMID- 17396297 TI - Re: contribution of 11C-choline positron emission tomography/computerized tomography to preoperative staging of advance transitional cell carcinoma. PMID- 17396298 TI - [Abstracts of the 34th Congress of the German Association for Rheumatology and the 20th meeting of the ARO, Wiesbaden, Germany, 18-21 October 2006]. PMID- 17396299 TI - Re: superficial (pT2a) and deep (pT2b) muscle invasion in pathological staging of bladder cancer following radical cystectomy. PMID- 17396300 TI - Abstracts of the 58th Congress of the German Society of Urology, Hamburg, Germany, 20-23 September 2006. PMID- 17396301 TI - [Diagnosis of hyperfunctioning adrenal masses]. PMID- 17396302 TI - Proceedings from the Workshop on the Prevention of Depression in Children and Adolescents, June 2004. PMID- 17396303 TI - Proceedings of the 2nd International Conference on Integrated Approaches to Sustain and Improve Plant Production under Drought Stress, Rome, Italy, 24-28 September 2005. PMID- 17396304 TI - Bevacizumab for non-small-cell lung cancer. PMID- 17396305 TI - Bevacizumab for non-small-cell lung cancer. PMID- 17396306 TI - Bevacizumab for non-small-cell lung cancer. PMID- 17396307 TI - Human H5N1 influenza. PMID- 17396308 TI - Human H5N1 influenza. PMID- 17396309 TI - [Lipoma in cerebellopontine angle]. PMID- 17396311 TI - Freedom to choose. PMID- 17396310 TI - [Recommendations for treatment of retraction pockets]. PMID- 17396312 TI - [The locum system is important in today's health services]. PMID- 17396313 TI - [We are fathers too, but...]. PMID- 17396314 TI - Selling a physician-owned hospital--15 key concepts. PMID- 17396318 TI - A puzzling link. Antidepressants and diabetes. PMID- 17396319 TI - Drink up! The wonder of water. PMID- 17396320 TI - Are you low? Hypoglycemia unawareness. PMID- 17396321 TI - Healthy eating. A small feast. Cooking for one or two. PMID- 17396323 TI - Your biggest fan. Need a little more support from your spouse or partner when it comes to your diabetes management? Here's how to talk about it. PMID- 17396322 TI - Hot shot. NBA rookie Adam Morrison scores with control. PMID- 17396324 TI - Fighting the epidemic. A United Nations resolution on diabetes. PMID- 17396325 TI - A kitchen in the tropics. Plan a winter getaway in the comfort of your own home. Recipes. PMID- 17396326 TI - Research profile. Low testosterone. Tough on insulin sensitivity. Frances J Hayes, MD. PMID- 17396327 TI - Policy watch. Using science, abusing science. PMID- 17396328 TI - Highlights of a year in AIDS. PMID- 17396329 TI - Health literacy, HIV and outcomes. PMID- 17396330 TI - Cytomegalovirus encephalitis in an HIV-seropositive person. AB - Cytomegalovirus (CMV) infection is common in persons with HIV/AIDS. It can affect the eye, lung, liver, GI tract, and nervous system. It is also a common cause of blindness in persons infected with HIV. We report a case of CMV encephalitis in a person in whom AIDS was recently diagnosed who did not have CMV retinitis on ophthalmological examination. PMID- 17396331 TI - Images in HIV/AIDS. HIV-associated lymphogranuloma venereum proctitis. PMID- 17396332 TI - Darunavir: an overview of an HIV protease inhibitor developed to overcome drug resistance. AB - Darunavir (TMC114) is a second-generation, sulfonamide, nonpeptidic protease inhibitor (PI) with a unique, flexible, 3-dimensional structure that contributes to it high potency and slow selection of resistant virus. Darunavir has to be coadministered with low-dose ritonavir and food to optimize its pharmacokinetics. Ritonavir-boosted darunavir is effective in many heavily pretreated patients, including those with multiple PI resistance mutations. In vitro, the coexistence of numerous PI mutations was required for its virological potency to be significantly reduced. Preliminary findings suggest that it is active against some tipranavir-resistant strains. PMID- 17396333 TI - Editorial comment: impact of darunavir for salvage therapy. PMID- 17396334 TI - Recent developments in HIV therapeutics. PMID- 17396335 TI - [Is there still a place for the correction of chronic venous insufficiency of the lower limbs by valvuloplasty]. AB - AIM: Various surgical procedures, extravenous (VPE) AND intravenous (VPI) valvuloplasties were described for the treatment of superficial (IVCS) and deep (IVCP) chronic venous insufficiency. METHODS: In 18 years: 153 VPE in limbs affected with IVCS. 9 VPE, 26 VPI (10 typical, 8 atypical, 8 autologous) and 38 other procedures in 74 limbs with IVCP selected and controlled by US (hemodynamic improvement or normalization = MNE) and X-ray (5 VPI). RESULTS: IVCS: 153 VPE=8,2% of the general casuistry. ;: MNE) 90,2% ( mean follow-up. 5 y,4 m.); stable at 10 y 68%; best results in IVCS <20 y. (Chi-sq.P=0.05). IVCP: VPE=12,3% and VPI=35,6% of the interventions; 9 VPE: MNE 88,8%; 26 VPI: MNE 81,4%; hemodynamic normalization by VPE-VPI(n.7/27=25,9%) (mean follow-up.7 y.,8 m.). CONCLUSIONS: VPE: Indicated in selected cases with IVCS and IVCP. In IVCP VPE-VPI are preferable than other procedures. Atypical and autologous VPI represents a technical progress in valveless syndromes. PMID- 17396336 TI - [Cardiovascular remodelling in patients with sub-clinical hypothyroidism]. AB - Sub-clinical hypothyroidism is defined by elevated serum thyroid-stimulating hormone level in the face of normal free thyroid hormone values. The role of sub clinical hypothyroidism as independent risk factor for atherosclerosis has been suggested by clinical studies which demonstrated a higher prevalence of peripheral arterial disease, aortic atherosclerosis and coronary artery disease, in patients affected by this pathological condition. This association have been confirmed by the assessment of subclinical atherosclerosis by means of B-mode ultrasonography. Using this method an higher intima-media thickness (IMT) of carotid artery, a close marker of early atherosclerosis changes, have been found in patients with subclinical hypothyroidism compared to control euthyroid subjects. Levothyroxine replacement therapy of sub-clinical hypothyroidism was able to improve both the carotid IMT and atherogenic lipid profile, suggesting that lipid infiltration of the endothelium may represent a mechanism underlying the atherosclerotic process in patients with this pathological condition. Morphologic and functional changes of the myocardial tissue has been also demonstrated in patients with sub-clinical hypothyroidism, using ultrasonic backscatter video densitometry. All these data provide evidence of cardiovascular remodelling in patients with sub-clinical hypothyroidism. Vascular remodelling in sub-clinical hypothyroidism patients could be also studied by means of backscatter analysis of carotid artery, a method which allows the assessment of vascular sclerosis. Our preliminary results using this method suggested that not only atherosclerosis by also sclerosis characterises vascular remodelling in sub clinical hypothyroidism patients. PMID- 17396337 TI - [Dysthyroidism and connective pathology]. AB - AIM: The aim of the study is to evaluate the degree of the relationship between dysthyroidism and connective pathology, both autoimmune diseases, presenting, sometimes, an early common symptom: the Raynaud phenomenon. METHOD: We studied 30 patients subdivided as follows: 18 affected by mixed connective pathology, 6 by scleroderma, 2 by CREST, 4 by scleroderma and Sjogren syndrome. We focused our attention on the chronology and the duration of the diagnosis, correlated to the laboratory parameters and the hypothesis of the pathogenetic pathway. RESULTS: 36.6% were positive to both the pathologies. One of the 11 patients resulted positive to hyperthyroidism and mixed connective, 10 patients resulted positive to hyperthyroidism subdivided in: 6 affected by sclerodermia and by 4 sclerodermia and S. of Sjogren. All the patients were positive to ANA without correlation between those values and the microsomal or antithyreoglobulin antibodies: the last two were present in 33%. The laboratory parameters indicating the inflammatory state were in the normal range, a light hypercholesterolemia was found in all patients. CONCLUSION: There is a suggestive relationship between the connective pathology and dysthyroidism,therefore it can be useful to evaluate always the association of both pathologies for the future identification of the immunogenetic alteration factors, the evaluation of the composition and the turnover of the collagen, subdividing the patients in relation to the clinical and the laboratory parameters, follow up to middle and long term. PMID- 17396338 TI - Plantar fasciitis. Help for heel pain. PMID- 17396339 TI - Health tips. Practicing relaxation. PMID- 17396340 TI - Antibiotic may improve irritable bowel syndrome. PMID- 17396341 TI - Myasthenia gravis. A treatable muscle weakness disorder. PMID- 17396342 TI - Colitis. A cause of persistent diarrhea in older adults. PMID- 17396343 TI - Because I have high blood pressure, my doctor has convinced me to quit reaching for the saltshaker at the dinner table. But what about using pepper? PMID- 17396344 TI - Mixed marks for heart surgery report cards. Grading heart surgeons on how well their patients to is a good idea, but the process still needs some work. PMID- 17396345 TI - 9 ways to protect your heart when diabetes threatens it. Diabetes and heart disease often go hand in hand. Here's how you can uncouple them. PMID- 17396346 TI - Aldosteronism: too much of a good thing. The hormone aldosterone helps the body manage water and sodium. Too much of it is an overlooked cause of high blood pressure. PMID- 17396347 TI - Statins for aortic valve narrowing? PMID- 17396348 TI - Different shades of gray for post-heart attack depression. Depression that develops for the very first time during recuperation from a heart attack affects recovery more than depression that started before the attack. PMID- 17396349 TI - Home defibrillator skill slip away. PMID- 17396350 TI - Ask the doctor. I know that exercise is good for my heart, which is one reason why I took up long-distance running. But I have heard that marathon running damages the heart. Is that true? PMID- 17396351 TI - Progress towards poliomyelitis eradication in Nigeria, January 2005 to December 2006. PMID- 17396352 TI - Epidemic alert and verification: summary report 2006. PMID- 17396353 TI - [Hot topics about surgical treatment of slow transit constipation]. PMID- 17396354 TI - [Abstracts of the 112th annual meeting of the Japanese Association of Anatomists. Osaka, Japan. March 27-29, 2007]. PMID- 17396355 TI - [The 95th annual meeting of the Japanese Urological Association. Kobe, Japan. April 14-17, 2007. Abstracts]. PMID- 17396356 TI - An objective method to assess the efficacy of theatre scissors. PMID- 17396357 TI - HIV-1 reverse transcriptase pausing at bulky 2' adducts is relieved by deletion of the RNase H domain. AB - Pausing by reverse transcriptase (RT) during retroviral replication increases the frequency of homologous strand transfer, nucleotide misincorporation, and non templated nucleotide addition. Pausing frequency increases at sites of DNA damage or upon incorporation of nucleotide analogs with steric barriers. These lesions thus likely stimulate mutations leading to resistant viral strains that escape drug treatments or immune surveillance. To study the response of retroviral RTs to bulky 2' adducts, a ribozyme-catalyzed reaction was used to generate an RNA template strand containing a thiophosphate adduct at a specific 2'-hydroxyl located upstream from a polyadenosine sequence. Subsequent alkylation increased the size of the adduct. Polymerization readthrough efficiencies were compared for mature RTs derived from HIV-1 (p66/p51), AMV (p95/p63), MMLV (p80 monomer), and a truncated version of HIV-1 RT lacking the RNase H domain (p51/p51 homodimer). Readthrough at the 2' lesion was markedly greater for the p51/p51 homodimer of HIV-1 RT than for the other enzymes, suggesting that the presence of the RNase H domain increases the probability that the modified primer/template will encounter a barrier to translocation. Comparison to published structures suggests potential unfavorable interactions between the 2' adduct and W24, F61, I63, D76, and R78 in the fingers domain of the RT. We propose that the enhanced readthrough observed upon RNase H domain deletion alters the trajectory of the primer/template in this region that diminishes steric and electrostatic clash with these residues. The template also included a penta-adenosine sequence that induced pausing in the order MMLV > HIV-1 (p66/p51) > AMV ~ HIV-1 (p51/p51). PMID- 17396358 TI - TAPP repair for inguinal hernias--a new composite mesh technique. PMID- 17396359 TI - Liver resection using the Atlas vessel sealing system. PMID- 17396360 TI - Commentary. A clinical outcome-based prospective study on venous thromboembolism after cancer surgery: the @RISTOS Project. PMID- 17396361 TI - Commentary. Risk factors for progression of peripheral arterial disease in large and small vessels. PMID- 17396362 TI - Commentary. Magnetic forces enable rapid endothelialization of synthetic vascular grafts. PMID- 17396363 TI - Commentary. Conservative versus surgical treatment of venous leg ulcers: a prospective, randomized, multicenter trial. PMID- 17396364 TI - Commentary. Protected carotid stenting in high-surgical-risk patients: the ARCHeR results. PMID- 17396365 TI - Use of a cortical screw to aid single piece excision of the trapezium: a simple technique. PMID- 17396366 TI - Kansas City focuses on animal health. New initiative to capitalize on cluster of companies and colleges. PMID- 17396367 TI - Equivocations of gender and rank: eighteenth-century sporting women. PMID- 17396368 TI - Round ligament fibroid. PMID- 17396369 TI - [The making of a "local society" in 19th-century northern England: Tyneside's "Great" boat race]. PMID- 17396370 TI - The Nottingham lead foot. PMID- 17396371 TI - Mimesis, materiality and the shadow of death. PMID- 17396372 TI - Competing allies: professionalisation and the hierarchy of science in Victorian Britain. PMID- 17396373 TI - The employment and retirement of older men, 1851-1881: further evidence from the census. PMID- 17396374 TI - "Sporty" girls and "artistic" boys: friendship, illicit sex, and the British "companionship" advertisement, 1913-1928. PMID- 17396375 TI - Chinese migrant women and families in Britain. PMID- 17396376 TI - Craniums, criminals, and the "cursed race": Italian anthropology in American racial thought, 1861-1924. PMID- 17396377 TI - [The historical production of places]. PMID- 17396379 TI - Renal mucinous and tubular spindle cell carcinoma: a clinicopathological study of 4 cases. PMID- 17396380 TI - What's your diagnosis? Adult with cough and bilateral ankle swelling. PMID- 17396381 TI - [Trends in early diagnosis of and therapy for schizophrenia]. PMID- 17396382 TI - [Trends in psychiatry]. PMID- 17396383 TI - [Present tasks and future aspects in psychiatric day care]. PMID- 17396384 TI - [Present tasks and future aspects in psychiatric day care service]. PMID- 17396385 TI - [Present tasks and future aspects in psychiatric day care]. PMID- 17396386 TI - [Present condition and tasks in the psychiatric day care section in a hospital]. PMID- 17396387 TI - [Pharmacological study on second-generation antipsychotic agents and the effects on improvement of cognitive function in patients with schizophrenia]. PMID- 17396388 TI - [Impaired cognitive function and presynaptic protein complexin in patients with schizophrenia]. PMID- 17396389 TI - [Progress in community mental health services and welfare in Wakkanai, Japan]. PMID- 17396390 TI - [Alcohol-related disorders]. PMID- 17396391 TI - Current world literature. Pathophysiology of hypertension. PMID- 17396392 TI - [Surgical treatment options for weight bearing articular surface defect]. AB - Treatment of full thickness defects of weight bearing articular surfaces is one of the most challenging problem of the daily orthopaedic practice. Recently published statistical evaluations of different arthroscopic findings support that 80% of full thickness defects of weight bearing areas leads to early osteoarthritis in a 5-10 years period. Beside traditional resurfacing procedures in the last two decades several new techniques have been developed to promote a hyaline or hyaline-like repair of the defected area. Basic science data, experimental results and preclinical evaluations supported wide clinical use of microfracture technique and autologous osteochondral mosaicplasty. Insurance company data confirm that yearly more, than 200 000 resurfacing procedures are performed according to the above mentioned two techniques all over the world. Beside these two popular resurfacing options autologous chondrocyte implantation may be a promising future alternative to treat extended defect areas. Beside basic science information and experimental data clinical experiences of more, than 8000 human implantations support such an application of cell therapy. This paper gives an outline of actual clinical resurfacing options and summarizes recent advances of cartilage repair. PMID- 17396393 TI - [The immunology of psoriasis: from basic research to the bedside]. AB - Psoriasis is a frequent, chronic, clinically variable inflammatory disease of unknown etiology, affecting primarily the skin and the joints. A cure for the disease is still missing, and due to the chronic course of the disease, currently available treatments are associated with serious morbidity. Psoriasis is considered to be an (auto)immune disorder, probably initiated by the overactive skin innate immune system, and maintained by immigrating activated type 1 T cells and abnormally proliferating and differentiating keratinocytes. A complex network of cytokines and chemokines mediates the pathological reaction, whereas the abnormal function of psoriatic regulatory T cells is likely responsible for the chronic nature of psoriasis. The most important clinical, histological, and pathogenic characteristics of psoriasis are discussed, and an overview of traditional and novel therapeutic modalities is provided. Based on recently obtained evidence from animal disease models and clinical studies using biological drugs with selective immunological action, a complex model for the immunopathogenesis of psoriasis is outlined. Advances in understanding the immunology of psoriasis have not only provided more insights into the cause and development of the disease, but gave new therapeutic tools into the hands of clinicians to more selectively and (possibly) more effectively treat psoriasis. PMID- 17396394 TI - [Short acting insulin analogues for treating diabetic patients with CSII (continuous subcutaneous insulin infusion)]. AB - The use of insulin pump treatment (CSII: continuous subcutaneous insulin infusion) became widely accepted in the last couple of years. A growing body of experiences accumulated in paediatric practice because CSII is preferable for treating young patients with type 1 diabetes. Nevertheless, CSII can be used, if indicated, for treating type 2 diabetic patients as well. Recently, fast acting insulin analogues are exclusively used for CSII. At moment, clinical observations with insulin lispro and insulin aspart are available but experiences with glulisine are still limited. Although some inconsistencies could be observed in the literature, it is widely accepted, that higher reduction in HbA(1c) values could be achieved by CSII as compared to intensive conservative insulin treatment; this could be more pronounced in cases with high initial HbA(1c) values. CSII with short acting insulin analogues could lead to a higher reduction of HbA(1c) values than CSII with human regular insulin. Moreover, the decrease of hypoglycaemic events could be expected in some cases. PMID- 17396395 TI - [The occurrence of malignancies in a Hungarian lupus population]. AB - INTRODUCTION: Chronic organ damages and complications have become one of the major problems concerning morbidity and mortality in systemic lupus erythematosus. AIMS: Present study was to examine the frequencies and types of malignancies in a lupus population followed regularly in our department, and to find correlation between cancer-associated mortality and the use of immune suppressants. METHODS AND RESULTS: Authors analysed data 860 lupus patients from 1965 till the end of 2004. During study period 164 patients died, 18 of whom from cancer. The frequency of cancer-associated mortality was 11% that was 2% of the total lupus population. Mortality has increased within 10-year periods reaching the maximum between 2001-2004 (8/24, 33%). Age of the 37 patients affected by malignancy was 47.7 (20-73) years. Cancers appeared 13.4 (1-45) years after the diagnosis of lupus. Breast cancer occurred most frequently (n=11) followed by gastro-intestinal malignancies (n=9), then cervix cancer and different hematologic malignancies (5-5 cases), bronchial cancer (n=4) and skin, ovarian and vesical cancers (1-1-1 cases). Relative risk of non-Hodgkin lymphoma and cervix cancer increased significantly as compared to those of the Hungarian general female population. Before the cancer developed 62.2% of patients received azathioprine and/or cyclophosphamide. CONCLUSION: Present results describe and analyse for the first time the association between lupus and malignant disorders in a Hungarian population calling the attention for the increased risk for the development of cancers and the importance of regular screening and extended examinations in SLE patients. PMID- 17396396 TI - [Veneral diseases in the Austro-Hungarian Army in the First World War]. PMID- 17396397 TI - [Ambulance in Nyiregyhaza before 1945]. PMID- 17396398 TI - Alcohol's effects on women's risk detection in a date-rape vignette. AB - Researchers have established that alcohol is a risk factor for date rape for both victims and perpetrators. OBJECTIVE: The authors tried to experimentally address the link between alcohol consumption and women's risk detection abilities in a risky sexual vignette. PARTICIPANTS: The authors recruited 42 women from undergraduate classrooms at a large midwestern university and randomly assigned them to drink an alcoholic (.04 blood alcohol content) or a placebo beverage. METHODS: Participants completed self-report inventories and listened to a date rape audiotaped vignette, which began with consensual sexual behavior and culminated in date rape, and the authors asked them to determine if and when the man should refrain from making further sexual advances. RESULTS: Student's t tests and Pearson r correlations showed that women who consumed alcohol and exhibited high levels of rape myth acceptance showed a significant decrease in risk recognition (p = .000 and .001, respectively). CONCLUSION: These findings highlight the significance of even small amounts of alcohol on behavior and cognition in women who are self-reported experienced drinkers. PMID- 17396399 TI - Spatial and temporal aspects of alcohol-related crime in a college town. AB - OBJECTIVE: The authors aimed to clarify crime "movement" through the city of Madison to focus efforts to address consequences of student drinking. The authors examined all crime reported by police during the 2003 year. METHODS: Using geographical information system (GIS) mapping and 2003 crime data from the University of Wisconsin-Madison and the city of Madison, the authors investigated the relationships among high-density alcohol outlets and various city neighborhoods. RESULTS: Different categories of crime showed different temporal and spatial patterns: Serious crimes peaked between 2 and 3AM, coinciding with bar closing time. Less serious crimes peaked between 11PM and midnight. Reports of vandalism peaked in the morning and afternoon. "Hotspots" for different crimes moved throughout the downtown. Results were consistent with problems associated with high-risk student drinking in college communities. CONCLUSION: These results and maps generated by GIS led directly to changes made by city and university officials to address high-risk drinking. PMID- 17396400 TI - Reasons for not reporting victimizations to the police: do they vary for physical and sexual incidents? AB - Victimization is a significant problem among college students, but it is less likely to be reported to the police than are victimizations in the general population. OBJECTIVE: In this study, the authors examined (1) whether reasons for not reporting varied by type of victimization (sexual or physical) and (2) victim-, offender-, and incident-related predictors of these reasons. PARTICIPANTS: To address these objectives, the authors used data collected from 492 female college students. METHODS: The authors recruited women via flyers placed around campus that asked them to come to the student health center to complete anonymous surveys. RESULTS: Findings from within-subject analyses indicated that women were more likely to cite the following reasons for not reporting a sexual rather than a physical victimization: the incident would be viewed as their fault, they were ashamed, they did not want anyone to know about the incident, or they did not want the police involved. Results from logistic regression analyses indicated that the predictors of not reporting also varied across crime types. CONCLUSIONS: The authors discuss study implications for campus-based prevention strategies. PMID- 17396401 TI - The effect of a college sexual assault prevention program on first-year students' victimization rates. AB - OBJECTIVE: Although a variety of sexual assault prevention programs are currently available to college health professionals, there is a dearth of information about the effect of these programs on sexual assault victimization rates. PARTICIPANTS: The authors evaluated the efficacy of a sexual assault prevention program for first-year students at a college in the Northeast (N = 1,982). METHODS: They used a retrospective cohort design and assessed the prevalence of sexual assault victimization among students exposed to the sexual assault prevention program and students 1 year their senior who were not exposed. RESULTS: Students who had no exposure were more likely to report that they were sexually assaulted during their first year of college (odds ratio = 1.74, 95% confidence interval [1.32 2.29]). Results suggest that the program was effective for males and females, but not for students with a prior history of sexual assault victimization. Gay, lesbian, and bisexual students were at increased risk for victimization as compared with heterosexual students, and students who drank alcohol or engaged in binge drinking were at increased risk as compared with alcohol abstinent students. CONCLUSIONS: Findings suggest that this program had a positive effect on victimization rates for certain sub-groups of students. PMID- 17396402 TI - Date fighting experiences among college students: are they associated with other health-risk behaviors? AB - OBJECTIVE: The authors examined the clustering of health-risk behaviors among college students who reported date fight involvement. PARTICIPANTS AND METHODS: The authors administered a Web-based survey to a stratified random sample of 3,920 college students from 10 universities in North Carolina. RESULTS: Among men, 5.6% reported date fight victimization, and 1% reported date fight perpetration. Victimization among men was associated with (1) first drink at age 15 years or younger, (2) a recent threat of violence by someone who had been drinking, (3) smoking, (4) amphetamine use, and (5) older age. Among women, 6.7% reported date fight victimization, which was associated with (1) older age, (2) assault from a student who had been drinking, (3) sex with 2 or more persons, (4) consumption of alcohol in high school, (5) illegal drug use, (6) nonsexual assault requiring medical treatment, and (7) living off campus. Of the women, 4.2% reported date fight perpetration, which was associated with (1) minority race/ethnicity, (2) older age, (3) frequency of sexual intercourse, and (4) alcohol and marijuana use. CONCLUSIONS: Date fight experiences were associated with multiple health-risk behaviors among this sample of college students. PMID- 17396403 TI - Keynote address: making campuses safer communities for students. PMID- 17396404 TI - Campus violence white paper. PMID- 17396405 TI - Coupling of photocatalytic and biological reactors to remove EDTA-Fe from aqueous solution. AB - Degradation of 2.5 mM EDTA-Fe solution was performed in a coupled photocatalytic biological reactor. The system consists of a photochemical annular reactor filled with TiO2 immobilized on glass Raschig rings coupled with an activated sludge continuous reactor. Around 50% of EDTA degradation was reached after 150 min irradiation. Simultaneously a four-fold increase in biodegradability, measured as BOD5/COD ratio, was observed. The activated sludge is not capable to degrade the complex EDTA-Fe but it removed partially the COD and efficiently the BOD5 of the photochemically treated solution. PMID- 17396406 TI - Optimization of nitrogen removal and start-up of Psyttalia sewage treatment works. AB - Bench scale experiments were conducted in order to finalize the design of the secondary treatment stage of Psyttalia Sewage Treatment Works that serves the Greater Athens area with a population equivalent of 3.5 million. Carbon and nitrogen removal efficiencies and activated sludge settling characteristics were determined through continuous flow experiments and determination of the maximum specific utilization rates of ammonia (AUR) and nitrate (NUR). According to the results of the pilot experiments the need arose for a significant increase in the initial design's anoxic volume to ensure a 70% average annual nitrogen removal. AUR experiments showed an insignificant inhibition to the nitrification process in the order of 0-30%, representative of typical domestic sewage. The findings of the pilot study were in good agreement with full scale data obtained during the start up of Psyttalia biological treatment plant. According to full scale data and AUR tests a very satisfactory nitrogen removal efficiency was obtained during start up. Finally both lab scale and full scale data indicated that the activated sludge plant operating on Psyttalia primary effluent that has a high oil and grease content, is prompt to experience foaming problems due to the excessive growth of filamentous microorganisms M. parvicella and Gordona amarae - like microorganisms. PMID- 17396407 TI - Inhibitory effect of a surfactant on pure cultures of a filamentous and a floc forming micro-organism. AB - Activated sludge is the most widely used biological process for wastewater treatment. Inorganic and organic compounds are removed by a biotic community in the aeration basin. Problems of these systems are loss of settleability and poor sludge compaction due to excessive growth of filamentous micro-organisms. The filamentous bulking can be controlled by the addition of chemical agents. Strong oxidants, such as chlorine, are utilized to eliminate filamentous bacteria; however, these substances also tend to attack floc-forming bacteria and to cause process breakdown. Besides, chlorine may become hazardous owing to the formation of chemical products as chloramines. Surfactant addition constitutes an interesting alternative for the control of filamentous bulking. In this work the effect of a surfactant Triton X-100 (octylphenol ethoxylate), on the respiratory activity (RA) of pure cultures of a filamentous (Sphaerotilus natans) and a floc former microorganism (Acinetobacter anitratus) was evaluated. In the concentration range tested (60-220 mg l(-1)), the surfactant was observed to exhibit high RA specific inhibition of the filamentous micro-organism with no significant effect on the floc-forming bacteria. Light microscopy observations showed that the surfactant induced cell lysis, leaving only empty sheaths in the case of filamentous micro-organisms. A kinetic equation to predict the microbial RA fraction of a S. natans pure culture as a function of surfactant concentration and contact time was proposed. The effect of Triton X-100 on the inactivation of pure cultures of both micro-organisms was compared to that of chlorine. Triton X 100 results were adequate to eliminate filamentous bacteria emerging as an alternative for filamentous bulking treatment. PMID- 17396408 TI - Toxicity evaluation of the process effluent streams of a petrochemical industry. AB - The physico-chemical characteristics and the acute toxicity of several wastewater streams, generated in the industrial production of synthetic rubber, were determined. The acute toxicity was evaluated in bioassays using different organisms: Danio rerio (fish), Lactuca sativa (lettuce) and Brachionus calyciflorus (rotifer). The removal of toxicity attained in the industrial wastewater treatment plant was also determined upstream and downstream of the activated sludge process. The results obtained indicate that the critical streams in terms of acute toxicity are the effluents from the liquid polymer unit and the spent caustic butadiene washing stage. The biological treatment was able to partially remove the toxicity of the industrial wastewater. However, a residual toxicity level persisted in the biotreated wastewater. The results obtained with Lactuca sativa showed a high degree of reproducibility, using root length or germination index as evaluation parameters. The effect of volatile pollutants on the toxicity results obtained with lettuce seeds was assessed, using ethanol as a model compound. Modifications on the assay procedure were proposed. A strong correlation between the toxic responses of Lactuca sativa and Danio rerio was observed for most industrial effluent streams. PMID- 17396409 TI - Potential for enhancement of aerobic biological removal of recalcitrant organic matter in bleached pulp mill effluents. AB - Increasingly stringent effluent quality limits for bleached kraft pulp mills pose a great challenge to mill wastewater system managers since these limits can require levels of chemical oxygen demand (COD) removal efficiency rarely reported for biological treatment of these types of effluents. The present study was therefore undertaken to better understand the nature of recalcitrant COD in bleached kraft pulp effluents that persists through the biological treatment system. Bleaching effluents from a Brazilian eucalypt bleached kraft pulp mill were collected and treated in a bench-scale sequencing batch reactor. Organic matter in raw and treated effluents was characterized before and after separation into low and high molecular mass fractions. Biological treatment removed 71% of the COD, with 83% removal of the low molecular mass COD but only 36% removal of the high molecular mass COD. Microorganisms capable of degrading the recalcitrant COD were isolated from enrichment cultures of the original activated sludge fed on fractions of the bleaching effluent that presented low biodegradabilities. Use of a microbial consortium composed of ten of these isolates to treat the biologically treated effluent removed a further 12% of the effluent COD, all from the high molecular mass fraction. Results of this research indicate that microorganisms with potential for degrading recalcitrant COD are present in activated sludge, but that these are not metabolically active during normal activated sludge treatment of mill effluents. The use of biological selectors in the treatment system to promote growth of such microorganisms may enhance removal of recalcitrant organic matter. PMID- 17396410 TI - Is phosphorus recovery from waste water feasible? AB - Phosphorus (P) recovery from waste water must become a predominant goal of all countries to face the limited resources of this essential nutrient. The induced crystallisation of calcium phosphates straight from the waste water phase applying tobermorite-rich calcium silicate hydrate compounds (CSH) from the construction industry as the trigger material has proved to be a suitable method. Laboratory and semi-technical scale experiments were carried out in fixed bed, stirred reactor and expanded bed mode. P-loads of the crystallisation substrates of up to 13 wt-% total P (P-tot) (30 wt-% P2O5) were achieved. Recycling options of the generated products, both as substitute for phosphate rock in the phosphate industry and as a new fertiliser in agriculture, were demonstrated. Indicative operating and investment costs were estimated for conversion of conventional waste water treatment plants (WWTP) designed for nutrient removal and P precipitation with iron and aluminium reagents to the proposed new crystallisation technology for simultaneous P-removal and P-recovery. PMID- 17396411 TI - Comparison of reject water treatment with nitrification/denitrification via nitrite in SBR and SHARON chemostat process. AB - A comparison between three feasible ways of developing Biological Nitrogen Removal (BNR) via nitrite to treat real reject water of 800-900 mg NH4(+)-N l(-1) is proposed. A Sequencing Batch Reactor (SBR) and a chemostat SHARON (Single reactor High activity Ammonium Removal Over Nitrite) continuous reactor were operated. In the SBR operation 0.8 kg N (d m3)(-1) was achieved, whereas in SHARON/denitrification the removal reached was 0.4 kg N (d m3)(-1). SHARON was also developed with partial nitrification of ammonium in order to obtain a stream ready for Anammox (Anaerobic Ammonia Oxidation) process obtaining an effluent with an average composition of 400 mg NO2(-)-N l(-1) and 350 mg NH4(-)-N l(-1). PMID- 17396412 TI - Oxygen uptake rate measurements for evaluation of ozonation of municipal wastewater. AB - Ozonation has become more frequently used as oxidant to reduce chemical oxygen demand (COD) and toxic substances in wastewater originating from different industrial processes. An ozonation pilot plant followed by two parallel biological filters are used to investigate the possibility to reduce the high concentration of refractory COD in the effluent from a municipal wastewater treatment plant, partly loaded with biologically treated wastewater from a large pharmaceutical industry. COD measurements are used to evaluate the overall reduction of organic matter, oxygen uptake rate (OUR) and volatile fatty acids measurements are used for evaluation of the biodegradability of the remaining COD after ozonation. The impact of the ozone dose on the overall COD degradation and degradability of the remaining COD has been estimated. Biochemical oxygen demand (BOD) measurements confirm that parts of the COD are converted into biodegradable organic matter by ozonation. Biofilters following the ozonation plant secure that any degradable organic matter produced by ozonation is removed, which is confirmed from OUR-measurements. PMID- 17396413 TI - Model-based design of a control strategy for optimal start-up of a high-strength nitrification system. AB - The objective of this study was to define an automatic control loop for the start up of a high-strength nitrification system to achieve a rich nitrifying biomass from a poor nitrifying sludge by means of simulation tools. The used model considered the nitrification as a two-step model with substrate and non competitive inhibitions. Two control strategies (on-off controller and proportional-integral (PI) controller) were designed, simulated and compared. The measured variable in the control loops was the sum of ammonium and nitrite concentrations in the effluent and the manipulated variable was the inflow. The objective in the optimisation of both controllers was to increase the inflow of the system as fast as possible without exceeding the maximum ammonium and nitrite concentrations allowed in the effluent. The optimised controllers parameters were used to simulate a 40 days start-up. The results obtained with both strategies were similar, although, the best strategy was the PI controller since it was less oscillatory and the biomass growth was slightly faster. PMID- 17396414 TI - Determination of suitable pretreatment method for old-intermediate landfill leachate. AB - For old-intermediate landfill leachate, the high concentrations of ammonium and low BOD/COD (B/C) ratios are the main obstacles to biological treatment. Magnesium ammonium phosphate (MAP) precipitation (struvite precipitation) and Fenton oxidation were employed as pretreatment options on this type of leachate. Results revealed that at optimum operating conditions of pH 9 and MAP ratio of 1:1:1, ammonium and COD were reduced by 97.8% from 2132 mg l(-1) to 46 mg l(-1) and by 50.0% from 4113 mg l(-1) to 2057 mg l(-1), respectively using struvite precipitation. COD was lowered by 76% and ammonium by 6.3% using Fenton oxidation at optimized conditions of H2O2 dosage of 3500 mg l(-1), Fe2+ dosage of 437.5 mg l(-1), pH 4, a mass ratio of 8 for H2O2: Fe2+ dosage and reaction time of 30 minutes. At their respective operating conditions, the B/C ratios for both pre treatment methods were increased from 0.35 in the raw leachate to 0.66 and 0.71, for MAP precipitation and Fenton oxidation respectively, values that are suitable for biological treatment. This study shows that struvite precipitation is a suitable pretreatment option resulting in the simultaneous removal of ammonium and organic matter in old-intermediate landfill leachate. PMID- 17396415 TI - Modeling contaminant transport in a three-phase groundwater system with the Freundlich-type retardation factor. AB - Colloid-facilitated contaminant transport was simulated in this study for the three-phase groundwater system where one or more sorption processes can be described with nonlinear sorption isotherm (Freundlich isotherm). A concise form of contaminant transport equation was derived from the mass balance equation of the contaminant. The developed model was numerically solved by the finite difference method along with the Picard iteration. The simulation results were used to quantitatively analyze the previously reported column data showing nonlinear sorption behavior. The analysis led to the following observations: (i) increases of the distribution coefficient of contaminant between the aqueous and solid phases (K(S)c) and the one between the dissolved natural organic matters and solid phase ( K(S)OM) generate less facilitation (i.e., late arrival of contaminant breakthrough curves (BTCs), and the distribution coefficient of contaminant between the aqueous and the solid phases (K(OM)c) gives the opposite result; (ii) the increase of the Freundlich constant for the sorption isotherm between the aqueous and the solid phases (N(S)c) yields the late arrival of BTC, and the other two Freundlich constants produce the opposite results; (iii) the Freundlich constants generally yield a sharper front as the BTC arrives at later pore volumes, while the distribution coefficients generally yield a more spread of the BTC as it arrives at later volumes. This modeling study shows that transport modeling provides a more efficient analyzing tool than the retardation factor alone concerning the colloid-facilitated contaminant transport with nonlinear sorption processes. PMID- 17396416 TI - Formation and hydrodynamic characteristics of aerobic granules in an activated sludge system. AB - Development of aerobic granules in the aeration tank of an activated sludge system has been studied. The introduction of activated carbon particles into the aeration tank resulted in the formation of biogranules containing activated carbon as core nuclei. The presence of activated carbon also induced the formation of self-immobilized granules, which did not have any carrier particle at their core. The presence of aerobic granules enhanced the treatment efficiency of the reactor. At an organic loading rate of 32.8 kg COD m(-3)d(-1) and 0.78 h hydraulic retention time (HRT), the reactor showed 96% COD removal efficiency. At an HRT of 0.272 h and organic loading rate of 46.7 kg COD m(-3)d(-1), the reactor outlet COD remained below 100 mg l(-1). Settling velocity studies carried out on the biogranules showed that the drag coefficient of biogranules is greater than that of the rigid particle at the same Reynolds number. PMID- 17396417 TI - Coupling of abiotic and biotic parameters to evaluate performance of combining natural lagooning and use of two sand filters in the treatment of landfill leachates. AB - A study in the Etueffont landfill, located in Belfort (France), was conducted to evaluate the performance of combining natural lagooning and use of two sand filters for treating leachates through the coupling estimation of several abiotic and biotic parameters. Two gravel filters were installed in the upstream of the first basin which communicates with the remaing 2, 3 and 4 basins. The distribution of physical-chemical (T, pH, Eh, EC, O2, SM, SO4(2-), Cl-, Zn, Fe, Mg, Ni, Al, As, Ba, Cu, Sn, Zn, BOD, COD, KN, NH4+, NO2+ ,TP, AOX: absorbable organic halides, VFA: volatile fatty acids, and atrazine) and biological (bacteria, protozoa, phytoplankton) parameters was assessed in the leachate entering in basin 1, and downstream of the filters. The results showed slight variations in the physical-chemical composition of the leachate between 1999 and 2000, most likely ascribed to the maturation of the landfill but a very significant removal of SM (suspended matter) by the sand filters. This, applied to the majority of the studied parameters. Thus, the sand filter treatment of the leachates combined with natural lagooning was efficient in the improvement of water clarification. PMID- 17396418 TI - Effect of rotifers on the stability of aerobic granules. AB - The current study evaluated the effect of rotifers on the stability of aerobic granules. Two sequence batch reactors (SBRs) with airflow rates of 4 (R1) and 6 (R2) 1 min(-1), respectively, were used to develop aerobic granules. Granules were well developed with excellent settleability in terms of SVI30 (sludge volume index,) of about 50 ml g(-1) in both reactors at the beginning. With the outgrowth of rotifers, granules completely disintegrated in R1 around cycle 500 (a cycle was 3 hours). However, after the rotifers disappeared, i.e. cycle 550, granules re-appeared with a slow settling rate in R1 (SVI30: 200-300 ml g(-1)). The rotifers mechanically damaged the structure of granules, resulting in disintegration. However, granules developed under high shear force seem to have strong resistance to rotifers. During re-granulation, a long time lag between the improvements of morphology and settleability suggested that re-granulation resulted from entanglement more than bio-attachment or bio-growth. Additionally, it was confirmed that the ratio of carbohydrate to protein extracellular polymeric substances (EPS) could well indicate the strength of granules. Protein EPS well correlated with the difference between SVI5 and SVI30 in R1, therefore, decreasing protein EPS would increase the compactness of granules. PMID- 17396419 TI - Data 'so poor it had to be disregarded'. PMID- 17396420 TI - Making an entrance. A new 86 million pound state-of-the-art treatment centre in Liverpool incorporates the design ideas of senior nursing and medical staff. AB - Broadgreen Hospital in Liverpool has undergone a transformation, creating a treatment unit that is both patient-friendly and enjoyed by the staff. PMID- 17396421 TI - Edge of endurance. A study of how climbers on Everest cope with low oxygen levels could benefit patients in intensive care. AB - A team of scientists and technicians will spend three months near Everest researching human performance at extreme altitude. Nurse Kay Mitchell is helping run the pioneering study. PMID- 17396423 TI - Why obesity is the new smoking. PMID- 17396422 TI - Waiting to practise. Many Zambian nurses come to work in the UK but face a struggle to find jobs that match their skills and experience. AB - Although there is an ethical agreement that the NHS will not recruit nurses from developing countries, many still come and work in the independent sector--often without their expertise being recognised. PMID- 17396424 TI - Where's the logic? Round-the-clock surgery could stretch hospitals to breaking point. PMID- 17396425 TI - Nurses' involvement in international research collaborations. AB - Nurses are becoming increasingly involved in collaborative international research teams. This article describes a European-wide research consortium that is implementing research across Europe. The experiences of members of a cross national research consortium and some of the challenges are discussed. Cross national research consortia should pay attention to preparatory, methodological, financial, operational and structural challenges. Ownership of data and dissemination of research findings are also explored. Consortia need to find a balance between a consistent organisational structure and communication styles, and respecting the differences between members of the consortium. PMID- 17396426 TI - Attitudes to using the laryngeal mask airway in paediatric resuscitation. AB - AIM: To explore nurses' attitudes towards, and confidence in, the use of laryngeal mask airway (LMA) and oral Guedel airway (OGA). The research formed part of a larger study which examined whether nurses could deliver adequate ventilation in children using these two airways compared to a consultant anaesthetist. METHOD: Twenty five critical care nurses were trained to insert an LMA and an OGA. They were then required to insert each airway into an unconscious apnoeic child in the anaesthetic room. A questionnaire was devised to ascertain the participants' preferred airway and their attitudes towards using the airways at various times during the study period. FINDINGS: Before training the nurses were evenly divided about which airway they preferred but this changed to favouring the LMA after training. However, by the end of the study the majority of participants preferred to use the OGA. CONCLUSION: The nurses' confidence increased as a result of practising these ventilating techniques on patients. They felt that being able to insert an LMA as a second-line airway when the OGA was not providing adequate ventilation was useful. However, they also felt that they would need continuous training to update this skill. PMID- 17396427 TI - Management of patients with asthma and allergic rhinitis. AB - This article discusses the pathophysiology and management of patients with allergic rhinitis. The author relates the condition to asthma, which is often experienced by those with allergic rhinitis. PMID- 17396428 TI - Nursing care of vascular disorders. PMID- 17396429 TI - The ride of your life. Tough, but incredibly rewarding, a nursing course is a life-changing event. PMID- 17396430 TI - Keeping to the same old pathways. Seven years after coming up with radical proposals to demolish professional boundaries the government has retired defeated. PMID- 17396431 TI - Taking the long view. Carol Webley-Brown looks at a new role in managing chronic conditions. PMID- 17396432 TI - Should off-label drug use be off-the-table? PMID- 17396433 TI - Role of physical exercise in children and adolescents with diabetes mellitus. AB - During the past 50 years several studies have underlined the central role of physical exercise in the management of patients with both type 1 and type 2 diabetes mellitus. The numerous benefits described in normal individuals who practise regular exercise have also been demonstrated in patients with diabetes who obtained significant physical and psychological advantages for the care of the underlying disease. Despite physical and psychological benefits, the occurrence of acute complications and some important effects on diabetes-related vascular complications may often discourage patients from participation in sports activities. However, even though adverse events may occur, exercise is still judged one of the most important components in the treatment of patients with diabetes. Thus, children, adolescents and young adults with diabetes must be educated on the metabolic changes occurring during physical activity in order to be able to acquire the ability to individually modulate their diet and insulin therapy before and after exercise. Appropriate education may allow a proper and correct approach to physical exercise. PMID- 17396434 TI - Transient, isolated growth hormone deficiency after a BB gunshot wound. PMID- 17396435 TI - Assessment of insulin sensitivity from measurements in fasting state and during an oral glucose tolerance test in obese children. AB - BACKGROUND: Few previous studies have examined the validity of the fasting glucose-to-insulin ratio (FGIR), homeostasis model assessment of insulin resistance (HOMA-IR) and quantitative insulin-sensitivity check index (QUICKI) in pediatric populations. OBJECTIVE: To compare simple indices of insulin resistance calculated from fasting glucose and insulin levels with insulin sensitivity indices (area under the response curve [AUCinsulin], insulin sensitivity index [ISI-compositeL) determined by oral glucose tolerance testing (OGTT) in obese children. METHODS: One hundred and forty-eight obese children and adolescents (86 girls and 62 boys, mean age: 10.86 +/- 3.08 years, mean body mass index (BMI): 27.7 +/- 4.2) participated in the study. OGTT was performed in all participants. After glucose and insulin measurements from OGTT, the children were divided into two groups according to the presence or absence of insulin resistance. Insulin sensitivity indices obtained from the OGTT were compared between the groups. The total plasma glucose response and insulin secretion were evaluated from the AUC estimated by the trapezoid rule. Cut-off points, and sensitivity and specificity calculations were based on insulin resistance with receiver operating characteristic curve (ROC) analysis. RESULTS: The prevalence of insulin resistance, glucose intolerance and dyslipidemia was 37.1%, 24.3% and 54% in obese children, respectively. The groups consisted of 93 children without insulin resistance (54 girls and 39 boys; mean age: 10.5 +/- 3.3 years; mean BMI: 27.0 +/ 4.2) and 55 children with insulin resistance (32 girls and 23 boys; mean age: 11.4 +/- 2.5 years; mean BMI: 27.9 +/- 3.9). There were significant differences in mean FGIR (10.0 +/- 7.2 vs 5.6 +/- 2.8, p < 0.001), HOMA-IR (3.2 +/- 2.3 vs 4.9 +/- 2.3, p < 0.001) and QUICKI (0.33 +/- 0.03 vs 0.30 +/- 0.02, p < 0.001) between the groups. The cut-off points for diagnosis of insulin resistance were < 5.6 for FGIR (sensitivity 61.8, specificity 76.3), > 2.7 for HOMA-IR (sensitivity 80, specificity 59.1), and < 0.328 for QUICKI (sensitivity 80, specificity 60.2). CONCLUSIONS: Indices derived from fasting samples for diagnosis of insulin sensitivity are reliable criteria in obese children and adolescents. HOMA-IR and QUICKI appeared to have similar sensitivity and specificity and to have higher sensitivity than FGIR. PMID- 17396436 TI - A community-based behavior modification intervention for childhood obesity. AB - Childhood obesity, caused by reduced physical activity and increased food consumption, has reached epidemic proportions. We hypothesized that a single practitioner could enable a child to reduce BMI by educating towards a healthier lifestyle and then reinforcing the message in a structured manner. In this study, intervention group participants and their parents received a half-hour talk on exercise and diet, repeated after 3 months. They were instructed to fill weekly diaries and were called weekly by telephone. Controls received the initial instruction only. Twenty-seven (14 intervention) obese children were recruited. Anthropometric parameters, fitness and biochemical data were collected before intervention and after 6 months in both groups. Sustained but not statistically significant improvements in attitude, BMI SDS and LDL-cholesterol were noted in the intervention group. These promising results support a need for further work to evaluate the efficacy and applicability of our approach in the population at large. PMID- 17396437 TI - Serum sialic acid, hs-CRP and oxidative stress parameters in obese children. AB - BACKGROUND: Obesity is a common health problem that is rapidly increasing among children. Obesity is accompanied by a high incidence of atherosclerosis, arterial hypertension and type 2 diabetes mellitus. We tested the hypothesis that the risk for atherosclerosis may be increased in obese children and measured some risk factors for atherosclerosis. PATIENTS AND METHODS: We determined sialic acid levels and investigated correlations with malondialdehyde (MDA), susceptibility to oxidation, total thiol concentrations, glucose and lipid profile in 39 obese (BMI 26.6 +/- 3.9) and 33 sex- and age-matched healthy children (BMI 15.9 +/- 1.7). RESULTS: MDA concentrations, susceptibility to oxidation and hs-CRP were significantly higher in obese children than controls. Sialic acid and total thiol concentrations were higher in controls but this did not reach statistical significance. Total cholesterol, LDL-cholesterol and glucose concentrations were significantly higher in the obese group and there was a positive correlation between BMI and MDA, susceptibility to oxidation, hs-CRP and glucose concentrations, and a negative correlation between BMI and HDL-cholesterol. Although sialic acid levels were not different between the groups, they showed a correlation with hs-CRP. CONCLUSIONS: A higher risk was found in obese children in relation to oxidative stress parameters, hs-CRP and lipid profile, and this risk showed a positive correlation with BMI. These results are important because children will encounter this increased risk for a longer time than adults, and taking care of obesity in childhood is especially important. PMID- 17396438 TI - Short stature in children with sickle cell anemia correlates with alterations in the IGF-I axis. AB - Children with sickle cell anemia (SCA) frequently have short stature. We propose that alterations in the IGF-I axis are involved in their growth failure. We investigated the IGF-I axis in children with SCA and height below the 25th percentile (n = 15) and compared it with that of children with SCA and height above the 50th percentile (n = 7). IGF-I and IGFBP-3 levels were assessed by RIA. IGFBP-3 proteolysis was assessed by a protease activity assay and by Western immunoblots. IGF-I and IGFBP-3 SDS were low for both groups. In the short statured patients, IGF-I SDS correlated with height velocity SDS (p = 0.018). IGFBP-3 SDS, when corrected for bone age, decreased with age (p = 0.0054). IGFBP 3 was proteolyzed in both groups although the short statured patients had lower levels of absolute intact IGFBP-3 when compared with the normally growing group (p = 0.028). We demonstrated that children with SCA have abnormalities in the IGF I axis, which worsen with age. PMID- 17396439 TI - Inappropriate use of potent topical glucocorticoids in infants. AB - Topical therapy with glucocorticoids (GCs) is used commonly in chronic dermatoses. Side effects are less common compared to systemic use; however, newer potent preparations may have serious side effects. A potential danger is their inappropriate use. Three infants who developed iatrogenic Cushing's syndrome and prolonged adrenal suppression in the course of GC therapy for simple diaper dermatitis are described. One patient also developed steatohepatitis which is uncommon with local GCs. PMID- 17396440 TI - Bone mineral status in children with congenital adrenal hyperplasia. AB - Congenital adrenal hyperplasia (CAH) is caused by a deficiency in an adrenal enzyme resulting in alterations in cortisol and aldosterone production. Bone status is affected by chronic glucocorticoid therapy and excess androgen exposure in children with CAH. This cross-sectional study enrolled participants with 21 hydroxylase deficiency from a pediatric referral center. Bone mineral density in the participants was normal when compared to age, gender and ethnicity adjusted standards, with respect to chronological age or bone age. Lean body mass was positively correlated with bone mineral content (BMC), independent of fat mass (p < 0.001). There was no significant correlation between glucocorticoid dose or serum androgen levels and skeletal endpoints. In conclusion, lean body mass appears to be an important correlate of BMC in patients with CAH. The normal bone status may be explained by the differential effects of glucocorticoids on growing bone, beneficial androgen effects, or other disease specific factors. PMID- 17396441 TI - The incidence and evolution of thyroid dysfunction during interferon-alpha therapy in children with chronic hepatitis B infection. AB - AIM: To assess the effect of interferon-alpha (IFN-alpha) therapy on thyroid functions in children with chronic hepatitis B infection (CHB). METHODS: Sixty eight children (7.8 +/- 3.6 years) were treated with 5 (n = 37, group I) or 10 MU/m2 (n = 31, group II) IFN for 6 months. Thyroid hormones, thyrotropin, thyrotropin-releasing hormone stimulation test, thyroid peroxidase and thyroglobulin autoantibodies were evaluated. RESULTS: Baseline features were not different in the two groups. After therapy, thyroid dysfunction was 27% and 41.9% in groups I and II (n.s.). Subclinical hypothyroidism was 17.9%/ 29%, subclinical hyperthyroidism 5.4%/12.9%, hypothyroidism 2.7%/-, and thyroid antibody positivity 2.7%/- in groups I and II (n.s.). Thyroid dysfunction was 33.8% in the whole group (p = 0.001). Predictors of IFN induced-thyroid dysfunction were female sex and age < 6 years. Thyroid dysfunction resolved within median 6 months in all but three children. CONCLUSION: Although IFN-induced thyroid dysfunction is mostly subclinical and reversible, this side effect should be kept in mind. PMID- 17396442 TI - Primary pigmented nodular adrenocortical disease (PPNAD) and pituitary adenoma in a boy with sporadic Carney complex due to a novel, de novo paternal PRKAR1A mutation (R96X). AB - We report the sporadic case of a 9 year-old boy with Carney syndrome, who presented with precocious puberty due to the endocrinological effects of primary pigmented nodular adrenocortical disease (PPNAD) and a synchronous pituitary adenoma. The adrenal tumor was removed surgically. Following unsuccessful treatment with bromocriptine the pituitary adenoma was also resected and a residual tumor irradiated. Thirty months after diagnosis the boy is free of symptoms. Mutation screening of the entire coding region of the PRKAR1A gene identified five single nucleotide exchanges, four of which were either heterozygous or homozygous polymorphic variants that were also present in his parents. However, the hitherto unreported disease-relevant mutation R96X in exon 3 had occurred de novo on the paternal allele. PMID- 17396443 TI - Hypocalcemia as a presenting feature of celiac disease in a patient with DiGeorge syndrome. PMID- 17396444 TI - Growth, growth hormone testing and response to growth hormone treatment in Gitelman syndrome. PMID- 17396445 TI - Evaluation of hypothalamic-pituitary-adrenal axis (HPA) suppression by low-dose (0.5 microg) and standard-dose (250 microg) adrenocorticotropic hormone (ACTH) tests in asthmatic children treated with inhaled corticosteroid (JPEM 2006; 19: 1015-1023). PMID- 17396446 TI - Something fishy. PMID- 17396447 TI - Think lean. PMID- 17396448 TI - At the centre of practice. PMID- 17396449 TI - Joining forces: Part 2. PMID- 17396450 TI - Ten steps to managing change. PMID- 17396451 TI - Towards a global nursing workforce: the 'brain circulation'. PMID- 17396452 TI - Leadership for service improvement: Part 2. PMID- 17396453 TI - [Therapeutic hypothermia after cardiac arrest seems to protect the brain. Treatment should be according to protocol, and treatment data registered]. PMID- 17396454 TI - [Specially designed CBT helps in somatization syndromes]. PMID- 17396455 TI - [Scientific controversies inevitable in the society of today. Important to learn form them--risk of contempt of knowledge among the general public]. PMID- 17396456 TI - [True or false findings? Application of Bayes principles can improve the assessment]. PMID- 17396457 TI - [The Swedish Resuscitation Council's guidelines to increase survival after cardiac arrest]. PMID- 17396458 TI - [Acute dyspnea]. PMID- 17396459 TI - [What is the therapeutic effect if the patient gets better, but no one knows how much better? Statistical method for paired ordinal data]. PMID- 17396460 TI - [Need for stronger unanimity for cooperation between psychiatry and consumers]. PMID- 17396461 TI - [The medical birth record facilitates longitudinal studies of the course of pregnancy]. PMID- 17396462 TI - [Well-founded role of HPV analysis in cervix cancer prevention]. PMID- 17396463 TI - [Misleading about patient injuries in primary health care]. PMID- 17396464 TI - [A conflict between compulsory reporting in health care and the European Union convention on human rights?]. PMID- 17396465 TI - [The world goes toward a warmer and richer future!]. PMID- 17396466 TI - [Overuse of nasal adrenergic agents--time for a warning, again!]. PMID- 17396467 TI - [The effect of pandemics on society. Historical experience necessary for today's preparedness]. PMID- 17396468 TI - ECG of the month. Abnormal electrocardiogram in a woman with a urinary tract infection. Sinus rhythm, rate 82/minute. Sagging ST segments, low T waves, and prominent U waves suggest hypokalemia. PMID- 17396469 TI - Radiology case of the month. Developmental delay. Agenesis of the corpus callosum. PMID- 17396470 TI - A 31-year-old, HIV-positive man presenting with emesis and bloody diarrhea. C. difficile infection. PMID- 17396471 TI - The epidemic of vitamin D deficiency. AB - A 37-year-old woman was referred to our endocrine clinic for management of her long-standing hypothyroidism. Her main complaints were muscle aches and pains that started about a year ago. The symptoms progressed to generalized muscle weakness. She described difficulty in getting out of her chair and in climbing stairs. She had an extensive work-up done by her neurologist and rheumatologist, including nerve-conduction studies and a muscle biopsy. The evaluation was normal, and she was diagnosed with fibromyalgia. She had gastric bypass surgery in 1998 and lost 150 pounds since the operation. She also has had lactose intolerance and a compression fracture without trauma. Her weight was 314 pounds. She had proximal muscle weakness. The rest of her physical examination was normal. Serum laboratory values are listed in Table 1. Based on the laboratory values, she was diagnosed as having severe vitamin D deficiency. She was started on 50,000 IU of vitamin D2 (Ergocalciferol) once a week for 6 months. Now her 25 hydroxyvitamin D level is 40 ng/mL, and her muscle strength has improved. PMID- 17396472 TI - Chagas disease in the United States: a cause for concern in Louisiana? AB - Chagas disease, or American trypanosomiasis, is an arthropod-borne protozoan infectious disease, hyperendemic throughout Latin America, caused by Trypanosoma cruzi, and transmitted to man by reduviid or kissing bugs. Throughout the Americas, Chagas disease shares many life cycle features with malaria, including transmission of infectious stages by local arthropods, exacerbation or reactivation of subclinical infections by immunosuppression (particularly HIV/AIDS) and pregnancy, and both transplacental and transfusion-related transmission. Although most cases of Chagas disease in the United States (US) are imported, significant numbers of Latin American immigrants contribute to the US blood supply and donate cadaveric tissues and organs for human transplantation, thus increasing the risks of both transfusion and transplantation-transmitted Chagas disease from unscreened blood products and transplantable tissues and organs. In addition, the risks of local reduviid bug-transmitted autochthonous or indigenous Chagas disease are also increasing as more immigrant workers enter the displaced populations of the Gulf South, including Louisiana, to assist in rebuilding efforts after Hurricanes Katrina and Rita. Although screening donated blood products for malaria remains impractical, consideration should now be given to serologically screening all donated blood and organs for Chagas disease in the US, as in most countries of Latin America, especially in high risk areas of California and the southern US, including Louisiana. PMID- 17396473 TI - Cleaner LASIK is possible. AB - LASIK (laser in situ keratomileusis) surgery is the most frequently performed refractive procedure for correcting nearsightedness, farsightedness, and astigmatism. Its popularity increased over the late 1990's, rapidly replacing Radial Keratotomy (RK). The LASIK operation and excimer laser surface ablation procedures gained the rapid acceptance of refractive physicians and patients over the RK procedure because of greater predictability, precision, safety, and stable vision. In the mid 90's, when doctors first began performing LASIK, it was referred to implicitly as the "FLAP and ZAP" procedure. This of course was an overly confident misstatement as unfamiliar complications began to appear. One of the ultimate goals in performing any surgical procedure is to minimize less than desirable outcomes arising from both infectious and non-infectious contaminants entering the surgical field. This is especially true of all corneal procedures, such as LASIK, where the normal mechanisms for fighting contaminations are diminished. PMID- 17396474 TI - Left vagus nerve stimulation for depression: first implantation case post-fDA approval and review of the literature. AB - OBJECTIVE: To describe the first case of left vagus nerve stimulation (VNS) following FDA approval and to review this in the context of existing literature. METHODS: A case report of the first patient undergoing VNS post-FDA approval is described. Indications, challenges, post-operative outcomes, and long-term implications are discussed. RESULTS: The patient treated had suicidal ideation, extensive alcoholism related to depression, and severely blunted affect with other major depressive symptoms. He failed multiple antidepressant medications, and over 50 electroconvulsive therapy treatments. Following stimulator implantation and subsequent stimulation, he had significant improvement over two weeks with respect to integration into his family, work, and abstinence from alcohol. In the context of these findings, existing literature on the use of VNS for treatment-resistant depression is reviewed. CONCLUSION: In selected cases, VNS has a significant role in treatment-resistant depression; the data suggest improvement in one-third of patients. Thus, they should be cautioned about side effects and should have a realistic level of expectation. PMID- 17396475 TI - Compartment syndrome secondary to spontaneous rupture of a Baker's cyst. AB - A 49-year-old woman with end-stage renal disease secondary to posterior urethral valves has received two kidney transplants since 1975, and both have succumbed to chronic rejection. She has been anuric since 2003 and undergoes hemodialysis three times a week. She was admitted to our hospital for evaluation for a third kidney transplant. The kidney was found to be unsuitable for this recipient, and she was taken to dialysis prior to discharge. Shortly after dialysis, she developed acute pain in the lower portion of her left leg and received a venous ultrasound to rule out possible deep vein thrombosis. No thrombus was appreciated. Instead, a Baker's cyst appeared to have ruptured its contents into her deep posterior compartment. In the twenty minutes it took to perform the ultrasound, her symptoms worsened, and her leg became firm. The patient reported extreme pain, paresthesias over the lateral aspect of the lower portion of the leg, and an inability to plantarflex or dorsiflex the foot. The foot was warm to the touch and still had a palpable pulse. The leg was beginning to lighten in color. The patient underwent an emergent fasciotomy. Pressure within the posterior compartment of the leg was measured at 120 mm Hg just prior to incision. The anterior, medial, and lateral compartments were measured at pressures of 23, 32, and 26 mm Hg, respectively. A two-incision anterolateral faciotomy was performed, and the wounds were left open to heal by secondary intention. The patient's convalescence was unremarkable, and she is still listed for renal transplantation. PMID- 17396476 TI - Aminocaproic-acid seal to reduce or prevent bleeding after liver biopsy. AB - This study investigates the efficacy of an aminocaproic-acid seal to prevent or reduce the risk of bleeding attendant to liver biopsies. The simple technique of occluding the biopsy tract by injecting 1-2 mL of aminocaproic acid, a fibrinolysis inhibitor, while withdrawing the biopsy sheath appears to reduce substantially the risk of delayed bleeding. The technique may be most useful if large core biopsy needles must be used to provide an adequate specimen. PMID- 17396477 TI - The Canadian ICU Collaborative: patient advocacy at its best. PMID- 17396478 TI - Roadblock to recovery: the surgical stress response. AB - Inadequately managed post-operative pain and the resulting surgical stress response (SSR) negatively affect patient outcomes. Critical care nurses need to understand that adequate pain management is critical to enabling patient recovery. A review of the physiology and pathophysiology of the SSR provides concrete evidence to substantiate the need for critical care nurses to prioritize nursing care that focuses on the prevention, early detection, and management of pain and the surgical stress response. Critical care nurses equipped with this evidence are capable of improving patient outcomes. PMID- 17396479 TI - The standardization of critical care nursing education and training: strategies for advancing clinical practice in Ontario's adult ICUs. AB - In 2004/2005, the Ontario Ministry of Health and Long-Term Care (MOHLTC) launched a critical care transformation strategy with a goal to enhance service delivery through improved access, quality and system resource management. Health human resources planning was seen as essential to the success of the strategy, particularly recruitment, education/training and retention of critical care nurses. A nursing task group was invited to articulate core competencies and practice standards that can be applied across Ontario's adult ICUs and to make recommendations for implementation and the training needed to encourage compliance with the initiative. In this article, the opportunity to position nursing within the Ontario MOHLTC vision is described, as well as the work undertaken to prepare for a province-wide approach to critical care nursing education and training. PMID- 17396480 TI - Neuromuscular blocking agents: enhancing safety by reducing the risk of accidental administration. AB - Neuromuscular blocking agents (NMBAs) are often found as ward stock in critical care units to ensure their availability in case of urgent need. The unintentional administration of an NMBA to a non-intubated and non-ventilated patient can result in severe permanent injury or death. Incidents involving mix-ups with NMBAs have occurred within and outside of critical care units. Case reports are highlighted with the intent to increase practitioner awareness of situations that could lead to similar errors and to promote changes in the critical care environment in order to enhance medication safety with NMBAs. PMID- 17396481 TI - Adjuvant treatment of non-small-cell lung cancer: how do we improve the cure rates further? AB - Surgery remains the initial treatment for patients with early-stage non-small cell lung cancer (NSCLC). Additional therapy is necessary because of high rates of distant and local disease recurrence after surgical resection. Early trials of adjuvant chemotherapy and postoperative radiation were often plagued by small patient sample size, inadequate surgical staging, and ineffective or antiquated treatment. A 1995 meta-analysis found a nonsignificant reduction in risk of death for postoperative cisplatin-based chemotherapy. Since then, a new generation of randomized phase III trials have been conducted, some of which have reported a benefit for chemotherapy in the adjuvant setting. The role of postoperative radiation therapy remains to be defined. It may not be beneficial in early-stage NSCLC but still may have utility in stage IIIA disease. Improvement in survival outcomes from adjuvant treatment are likely to result from the evaluation of novel agents, identification of tumor markers predictive of disease relapse, and definition of factors that determine sensitivity to therapeutic agents. Some of the molecularly targeted agents such as the angiogenesis and epidermal growth factor receptor inhibitors are being incorporated into clinical trials. Preliminary results with gene-expression profiles and lung cancer proteomics have been promising. These techniques may be used to create prediction models to identify patients at risk for disease relapse. Molecular markers such as ERCC1 may determine response to treatment. All of these innovations will hopefully increase cure rates for lung cancer patients by maximizing the efficacy of adjuvant therapy. PMID- 17396482 TI - Hormone-refractory prostate cancer: choosing the appropriate treatment option. AB - Hormone-refractory prostate cancer (HRCaP) is both heterogeneous and lethal. Multiple treatment options exist, including secondary hormonal manipulations, chemotherapy, experimental options, and best supportive care. Choosing the appropriate therapy for an individual patient depends on several important clinical factors such as the presence or absence of symptomatic metastatic disease, age and comorbidities, and prostate-specific antigen velocity. While only docetaxel (Taxotere)-based chemotherapy has been proven to improve survival in this setting, a wide range of therapies may be effective for any individual. Palliative maneuvers, such as external-beam radiation, bisphosphonate therapy, radiopharmaceuticals, and pain management are critical for appropriate patient management. Several promising novel therapies are in late-stage testing and will hopefully provide more treatment options for these patients. PMID- 17396483 TI - Novel concepts in radioimmunotherapy for non-Hodgkin's lymphoma. AB - Tositumomab/iodine-131 tositumomab (Bexxar) and ibritumomab tiuxetan (Zevalin) are radioimmunoconjugates targeting the CD20 antigen. Both agents are approved in the United States for use in relapsed or refractory, indolent or transformed, B cell lymphoma. These agents are well tolerated and have the highest levels of single-agent activity observed in these histologies. This review will summarize the key trials that led to approval of both I-131 tositumomab and ibritumomab tiuxetan, and then focus on four novel therapeutic concepts in radioimmunotherapy: retreatment, therapy of de novo indolent lymphoma, therapy of aggressive histologies, and incorporation in high-dose therapy programs utilizing autologous stem cell support. PMID- 17396484 TI - The role of intraperitoneal therapy in advanced ovarian cancer. AB - Intraperitoneal (i.p.) chemotherapy is a preferred treatment option that should be offered to all women for front-line treatment of stage III optimally debulked ovarian cancer. Patients should be provided with information on the survival and toxicity for both i.p. and intravenous (i.v.) therapies, as well as practical information about the administration of each regimen, so that they may play an active role in the decision-making process. When making a decision between i.p. and i.v. therapeutic options, the experience and preference of the oncologist are critical factors in determining appropriate therapy for each woman. PMID- 17396485 TI - Adult sarcomas: a missed opportunity for sustained cooperative group translational and clinical research. AB - Preclinical advances offer an opportunity to further reduce morbidity and mortality from sarcomas over the next decade. Since no single institution or North American cooperative oncology group has the expertise or patient resources for histology-specific clinical and translational research on adult sarcomas, efforts have been made to develop funding from the National Cancer Institute (NCI). One such initiative was the Intergroup Coalition Against Sarcomas (ICAS), which, building upon the strengths of the multimodality cooperative oncology groups, provided an infrastructure for broad participation by investigators from all treatment disciplines in protocol development and patient entry. However, despite an excellent evaluation in formal peer review, the Division of Cancer Treatment of NCI has ended this initiative claiming insufficient available funds- to the detriment of adult sarcoma patients now and in the future. PMID- 17396486 TI - PET imaging: update on sarcomas. PMID- 17396487 TI - Avoiding stress and burnout in cancer care. Words of wisdom from fellow oncologists. PMID- 17396488 TI - The time has come: describing and shaping RN practice. PMID- 17396489 TI - Bringing life-changing experiences home. PMID- 17396490 TI - Labels are libelous. PMID- 17396491 TI - [Compatible biomass estimation models of natural forests in Changbai Mountains based on forest inventory]. AB - Forest biomass estimation is the groundwork of analyzing carbon cycle and its dynamics in terrestrial ecosystems, but the current estimation models had the problem of un-compatibility between total forest biomass and its components. Taking the Wangqing natural forest area in Jilin Province as a case, the compatible concept of building forest biomass models based on forest invent' tory was brought forward. A series of compatible biomass estimation models, taking stem volume as one of the independent variables, were established by using simultaneous equations. The accuracy of the models in estimating the biomass of coniferous forest, broad-leaved forest, and mixed forest was all higher than 95% , suggesting that these models basically solved the problem of un-compatibility between total forest biomass and its components. PMID- 17396492 TI - [Species diversity and interspecific association in development sequence of Hippophae rhamnoides plantations in Loess hilly region]. AB - Based on field investigation, this paper analyzed the characteristics of species diversity and interspecific association at different development stages of Hippophcze rhamnoides plantations. The results showed that the species diversities of shrub layer, grass layer, and whole community of H. rharnnoides plantations were all fluctuated in "S" shape. At different development stages, the species richness and diversity were higher in grass layer than in shruh layer. The shrub species diversity was lower on hare land, but increased gradually with development stage. Shrub evenness index was higher in 13-year forest stand, while grass diversity index was higher in 3-year plantation, lower in 8-year plantation, and higher in 25-year plantation. The positive and negative absolute values of interspecific association between H. rharnnoides and other principal species changed in parabola shape, and the notable degree and the interspecific association intensity were weaker in 13-year plantation, showing that the species substitution rate was slower, competition was less, and community composition and its structure were relatively stable. To improve ecological environment, the H. rhamnoides plantations younger than 13 years old should he given priority to protection, while for those of 25 years old, moderate thinning should be made to promote the regeneration. PMID- 17396493 TI - [Photosynthetic physiological adaptabilities of Pinus tabulaeformis and Robinia pseudoacacia in the Loess Plateau]. AB - With Yangling, Yongshou, Fuxian, Ansai, Mizhi and Shenmu, the s ix counties from the south to the north in the Loess Plateau as study sites, this paper studied thoe photosynthetic charac teristics and leaf traits of Pinus tabulaeformis and Robinia pseudoacacia. The results showed that among the six sites, there were significant differences in the photosynthetic rate (Pn), photosynthetic nitrogen use efficiency (PNUE), water use efficiency (WUE), leaf mass per area (LMA), nitrogen content (Nmass), and chlorophyll content (Chl) of P. tabulaeformis and R. pseudoacacia, suggesting that the photosynthetic capacity and leaf traits of the two species differed with sites. From the south to the north, the Pn, PNUE and WUE of P. tabulaeformis increased slightly while those of R. pseudoacacia decreased significantly, indicating that in drought habitat, P. tabulaef6rmis could still maintain high photosynthetic capacity, hut the photosynthetic capacity of R. pseudoacacia was greatly restrained. Also from the south to the north, the LMA of P. tabulaeformis and R. pseudoacacia had a slight increasing trend, while Nmass and Chl decreased slightly. The variation ranges of the three parameters were greater for R. pseudoacacia than for P. tabulaeformis, indicating that P. tabulaeformis had stronger drought-tolerant capability than R. pseudoacacia, which was not only exhibited in physiological metabolism, but also in leaf morphological acclimation. The correlation analysis between photosynthetic parameters and leaf traits of P. tabulaeformis and R. pseudoacacia in the six sites showed that there was a significant negative correlation between LMA and Nmass. The Pn and PNUE of both test species had no correlations with LMA and Nmass, but had significant positive correlation with Chl. The WUE of the species was negatively correlated with LMA, but positively correlated with Nmass. PMID- 17396494 TI - [Effects of light intensity on seed germination and seedling early growth of Shorea wantianshuea]. AB - With shading experiment, this paper studied the seed germination and seedling early growth of Shorea wantianshuea, a representative species in Xishuangbanna seasonal rain forest under different light intensity and forest habitat. The results showed that both 100% light and deepest shading did not facilitate the germination of S. wantianshuea seed, while medium shading accelerated the seed germination. Gap center was more appropriate to the seed germination than gap edge and under-canopy. The seeds in gap center had the highest germination rate and rapid germination. Under partially shading, the seedling height, diameter of basal stem, and leaf area per seedling were in their peak, while the root-shoot ratio decreased with increasing shading and achieved its maximum under 100% light. The specific leaf area increased with increasing shading in a certain range, and peaked under 4.2% light. Gap center was beneficial to the early seedling growth, with all growth parameters except root-shoot ratio peaked there. The effects of environmental factors, mainly light intensity and quality, on the seed germination and seedling growth of S. wantianshuea were discussed. PMID- 17396495 TI - [Light response of Wisteria sinensis leaves physiological parameters under different soil moisture conditions]. AB - With two years old Wisteria sinensis as test material, this paper measured the light response of its leaves net photosynthetic rate (Pn), transpiration rate (Tr) and water use efficiency (WUE) under different soil moisture conditions, aimed to ascertain the optimal soil moisture and light conditions of W. sinensis growth. The results showed that the Pn, Tr and WUE had evident threshold responses to the variations of soil moisture and light intensity. To maintain the normal growth and higher levels P. and WUE of W. sinensis, soil volumetric moisture content (Wr) and relative moisture content (Wv) should be within the range of 15.3%-26.5% and 46.4%-80.3%, and the optimal Wv and Wr were 23.3% and 70.6%, respectively. W. sineasis leaves had stronger adaptability to light conditions. When the photosynthetic active radiation (PAR) was 600-1600 micromol x m(-2) x s(-1), the Pn and WUE were at higher level, and the light saturation points of Pn and WUE were all at about 800-1000 x micromol x m(-2) x s(-1). The appearance of non-stomatal limit was significantly correlated with soil moisture and light intensity. When W, ranged from 18.4% to 26.5%, the decline of Pn was mainly caused by stomatal limit rather than PAR. Out of this range, Pn was obviously affected by PAR, and the critical turning point of PAR was observed with the change from stomatal limit to non-stomatal limit. The minimal values of Wv and Wr for the normal growth of W. sinensis were 11.9% and 36.1%, respectively, and the maximal PAR was 1000 micromol x m(-2) x s(-1), the critical point of detriment in leaf photosynthetic organ. PMID- 17396496 TI - [Transpiration of banana plant measured by Granier method]. AB - Banana plant is the highest herbaceous plant in the world. Because of the height, it's inconvenient to measure the transpiration (Tr) of banana plant in direct way. Some indirect methods such as soil water balance method or FAO 56 Crop Coefficient are commonly used to measure or calculate the Tr, but the obtained results are easily influenced by soil texture and water content, climatic condition, and agronomic practice. In the present paper, Granier method was used to measure the sap flow (SF) at the corm of banana in a greenhouse from November 15 to October 5, 2005, with the Tr measured by gravimetric method compared. The results showed that the daily SF measured by Granier method was about 4% lower than the daily Tr measured by gravimetric method, and the SF rate lagged the Tr rate about one hour. Granier method was not sensitive enough to measure SF when the daily Tr was less than 0.05 L x m(-2) active leaf area. Granier probes could work properly 2-3 days after installation, and the measurement error of Granier method could be much reduced when more sets of probes were installed and the mean values of SF rate were taken. PMID- 17396497 TI - [Numerical simulation of vertical one-dimensional water movement in unsaturated soil: a case study on coniferous forest brown soil on northern slope of Changbai Mountains]. AB - This paper established a mathematical model to simulate the vertical one dimensional flow processes in unsaturated zone, based on the basic equation of soil water movement and the actual layer structure of coniferous forest brown soil on northern slope of Changbai Mountains. The numerical discrete equations of this model were derived, and the procedure for the numerical simulation of the model was programmed. This model was validated with numerical simulation, and the simulation values were consistent well with the measurements. PMID- 17396498 TI - [Seasonal dynamics of ecosystem services of grassland in Qinghai-Tibetan Plateau]. AB - Based on the theory of ecosystem services and by using RS and GIS techniques, the ecosystem services of grassland in Qinghai-Tibetan Plateau were analyzed and evaluated, with growth season as time unit. The results showed that the service of supplying biomass presented an up-down-up trend, and the peaks appeared in May and September. Its main affecting factors were PAR, soil moisture condition, and underground biomass. The service of carbon accumulation and oxygen release also presented up-down-up trend, which was related to the supplied biomass and its formation. As for the services of holding soil nutrients and water, the former had a peak in May, while the latter presented staggered variation due to the different water consumption characters of herbages at their different growth stages. PMID- 17396499 TI - [Effects of high field temperature on summer maize grain yield and quality]. AB - A field study with two summer maize cultivars ND108 and YD13 showed that increased field temperature (3 degrees C) at the stage from male tetrad to maturity had significant effects on their grain yield and quality. The grain yield of ND108 and YD13 decreased by 46.6% and 45.1%, grain crude protein content increased by 11.6% and 2.02%, ether extract content decreased by 38.9% and 9.6%, starch (mainly amylopection) content decreased by 7.0% and 8.4%, and lysine content decreased by 58.9% and 51.2%, respectively. PMID- 17396500 TI - [Characteristics of CO2, CH4 and N2O emissions from winter-fallowed paddy fields in hilly area of South China]. AB - With closed static chamber and modified gas chromatograph (HP5890 II), the in situ measurements were made on the CO2, CH4 and N2O emissions from winter fallowed paddy fields in the hilly area of South China. Gas samples were taken simultaneously from the fields with and without rice stubble. The results showed that both of the fields had the peak value of CO2 flux in the late afternoon. In the fields with and without rice stubble, the CH4 flux was positive in the day time while negative in the night, and the N2O flux in the day time was 1.79 and 1.58 times as much as that in the night, respectively. The diurnal average CO2 flux in the field with rice stubble was significantly higher than that in bare field (P < 0.05). Correlation analysis demonstrated that the CO2 flux in winter fallowed paddy fields had significant correlations with soil temperature, aboveground temperature, and air temperature, suggesting that temperature was the main factor affecting the CO2 emission from rice field after harvesting. During the observation time (from 2003-11-10 to 2004-01-18), the average CO2, CH4 and N2O fluxes in the field with rice stubble were (180.69 +/- 21.21) mg x m(-2) x h( 1), (-0.04 +/- 0.01) mg x m(-2) x h(-1) and (21.26 +/- 19.31) microg x m(-2) x h( 1), respectively. Compared with bare field, the CO2 flux in the field with rice stubble was 13.06% higher, CH4 absorption increased by 50%, while N2O flux was 60.75% lower. It was concluded that the winter fallowed paddy field in hilly area of South China was the source of atmospheric CO2 and N2O, and the sink of atmospheric CH4. PMID- 17396501 TI - [Heat stress characteristics of photosystem II in eggplant]. AB - With lower-and higher heat-resistant varieties of eggplant (Solanum melongena L.) Heibei I and Heibei II as test materials, and by using Plant Efficiency Analyzer (PEA) from Hansatech, this paper measured the fast chlorophyll a fluorescence transient and its parameters. The results showed that PS II construction became more sensitive to heat stress when ambient temperature was higher than 40 degrees C. The F0 went up slowly, and Fv/Fm and deltaF/Fm' came down dramatically. Heibei II had a longer semi-attenuation temperature of Fv/Fm (T50) and deltaF/Fm' (t50) than Heibei I. Under strong heat stress (5 min at 48 degrees C or 20-30 min at 44 degrees C), the K-step in relation to the inactivation of oxygen-evolving complex appeared in fluorescence rise at about 700 micros, and the regular O-J-I-P transient was transformed to O-K-J-I-P one. The K-phase of Heibei I and Heibei II appeared when the treatment time was up to 20 and 30 minutes at 44 degrees C, respectively. In comparing with 35 degrees C heat treatment, the DI0/RC in the parameters of Strasser's specific energy fluxes model was increased by a great extent under 48 degrees C or more heat stress, reflecting a strong safeguard of energy dissipation to PS II. When the temperature of heat stress increased from 35 degrees C to 52 degrees C, the Fvi/Fv of PS II silent reaction centers of Heibei I and Heibei II increased remarkably. PMID- 17396502 TI - [Effects of potassium application rate and time on the uptake and utilization of nitrogen and potassium by winter wheat]. AB - The study with 15N tracing techniques showed that basal application of K did not increase the N and K accumulation in wheat plant when the available K content in 0-20 cm soil layer was 118.5 mg x kg(-1), but increased the accumulation significantly when the soil available K content was 79.0 mg x kg(-1). When the fertilizer K was 1/2 basal-applied and 1/2 top-dressed at elongation stage, the absorbed fertilizer N and soil N by plant as well as the residual N of top dressed N in soil were increased with increasing K application rate, while the fertilizer N loss decreased. Compared with basal application, the split application of K increased the accumulated amount, uptake efficiency, and productive efficiency of N and K significantly. When the K application rate was 135 kg x hm(-2), split application of K promoted the absorption of top-dressed and soil N by plant, and increased the residual N of top-dressed N in soil. The results also showed that K application increased the grain yield and its protein and wet gluten contents. At the same K application rate, split application had better effect than basal application, and treatment K45 +45 (45 kg x hm(-2) as basal and 45 kg x hm(-2) top-dressed) showed the best effect. Applying excessive K tended to decrease wheat yield and its quality. PMID- 17396503 TI - [Effects of free-air CO2 enrichment (FACE) on yield formation of wheat]. AB - To investigate the effects of predicted higher CO2 levels on the growth duration, plant height, yield, and yield components of wheat (Triticum aestivum L.), a free air CO2 enrichment (FACE) experiment with weak gluten variety Ningmai 9 was conducted at Anzhen of Wuxi in Jiangsu Province in 2001-2002 and 2002-2003 growth seasons. The target [CO2] in FACE plots was 200 microl x L(-1) above that in ambient air. Three levels of N were supplied, i.e., 90 kg x hm(-2) (2001-2002) and 125 kg x hm(-2)(2002-2003) (low level, LN), 180 kg x hm(-2) 2002-2003) (medium level, MN), and 250 kg x hm(-2)(high level, HN). The durations from sowing to heading and from heading to maturity and the whole growth period of wheat in FACE plots shortened 1.3 , 1.3 and 2.6 days, respectively, compared with the control. FACE increased the plant height (+4.0% significantly, due to the increases of panicle length and of the first and second internode lengths. FACE also greatly increased the grain yield by an average of 24.6%. Across the two years, there was a positive [CO2] x N interaction for grain yield, with a yield increase of 15.2%, 21.4% and 35.4% at LN, MN and HN, respectively. The ears per square meter in FACE plots was increased by an average of 17.8% mainly due to the increase of maximum tiller number per unit ground area rather than that of the percentage of productive tiller (panicle bearing). In addition, FACE increased the grain number per ear (+2.9% and the individual grain mass (+4.8%). PMID- 17396504 TI - [Allelopathy of garlic root exudates on different receiver vegetables]. AB - By the method of tissue culture under sterilized condition, this paper studied the allelopathy of garlic root exudates on lettuce, hot pepper, radish, cucumber, Chinese cabbage, and tomato. The results showed that garlic root exudates had no evident effects on the germination rate, germination index, shoot height, and protective enzyme system of test crops, but significantly increased the root length, aboveground fresh mass, and root fresh mass of lettuce, with the RIs being +0.163, +0.106, +0.318, respectively. The exudates also increased the root length of Chinese cabbage, with a RI of +0.120. For other test crops, no significant difference was observed between the treatments and the control. Garlic root exudates significantly increased the chlorophyll content and root activity of the receiver vegetables. The strongest promotion effects were found on chlorophyll content in radish, with RI being +0.282, and on root activity of cucumber, with RI being +0.184. The exudates promoted the nutrient absorption of all the receiver vegetables. PMID- 17396505 TI - [Effects of elevated CO2 on photoinhibition of strawberry leaves under different nitrogen levels]. AB - By using PAM-2000 portable chlorophyll fluorometer and HCM-1000 photosynthesis measurement system, this paper measured the initial fluorescence (Fo), maximal photochemical efficiency of PSII (Fv/Fm), maximal fluorescence (Fm), amount of inactive PS II reaction centers (Fi-Fo), proportion of Q(B)-non-reducing PS II reaction centers [(Fi-Fo)/(Fp-Fo)], and net photosynthetic rate (Pn) of strawberry leaves under conditions of elevated CO2 (700 microl x L(-1)) and ambient CO2 (390 microl x L(-1)) at three levels of nitrogen application (12, 4 and 0.4 mmol x L(-1)). The results showed that there was a significant joint effect between CO2 and N on the photoinhibition of strawberry leaves. Under elevated CO2 condition, the Pn in treatment 12 mmol N x L(-1) increased by 62.7%, while that in treatments 4 and 0.4 mmol N x L(-1) decreased by 7.4% and 21.3%, respectively. When exposed to high light and subsequently recovered in dark for 4 hours, the strawberry leaves in treatment 12 mmol N x L(-1) showed less changes of Fm and Fv/Fm in elevated CO2 than in ambient CO2, while those in treatments 4 and 0.4 mmol N x L(-1) were in adverse, suggesting that for the strawberry leaves in elevated CO2, nitrogen deficiency could result in an acclimatized decrease of photosynthesis and an increase of photoinhibition. PMID- 17396506 TI - [Desorption characteristics of inorganic nitrogen in vegetable garden soil and their effects on soil NO3- -N loss potential]. AB - With pot experiment and soil nitrogen desorption model, this paper studied the characteristics of nitrogen desorption in vegetable garden soil, and their effects on the NO3- -N concentration of soil leachate. The results showed that soil leachate NO3- -N concentration had a non-linear relationship with the parameters Q, Cli and C1/lamda of soil nitrogen, but the relationship became linear when these eigenvalues were relatively low. A conception of hi-curve cross point was put forward to assess the soil NO3- -N loss potential. When the eigenvalues were higher than the hi-curve cross point X0, the NO3- -N concentration in soil leachate would be increased rapidly in non-linear form, while on the contrary, the increase would be maintained at a lower level. PMID- 17396507 TI - [Nutrient contents and microbial populations of aeolian sandy soil in Sanjiangyuan region of Qinghai Province]. AB - Sanjiangyuan region (the headstream of three rivers) in Qinghai Province of China is the highest and largest inland alpine wetland in the world. The study on the nutrient contents and microbial populations of aeolian sandy soils in this region showed that soil organic matter content increased with the evolution of aeolian sand dunes from un-stabilized to stabilized state, being 5.9 and 3.8 times higher in stabilized sand dune than in mobile and semi-stabilized sand dunes, respectively. Soil nitrogen and phosphorus contents increased in line with the amount of organic matter, while potassium content and pH value varied slightly. The microbial populations changed markedly with the development of vegetation, fixing of mobile sand, and increase of soil nutrients. The quantities of soil bacteria, fungi and actinomycetes were 4.0 and 2.8 times, 19.6 and 6.3 times, and 12.4 and 2.6 times higher in stabilized and semi-stabilized sand dunes than in mobile sand dune, respectively, indicating that soil microbial bio-diversity was increased with the evolution of aeolian sand dunes from mobile to stabilized state. In addition, the quantities of soil microbes were closely correlated with the contents of soil organic matter, total nitrogen, and available nitrogen and phosphorus, but not correlated with soil total phosphorus, total and available potassium, or pH value. PMID- 17396508 TI - [High effective phosphate-solubilizing bacteria: their isolation and promoting effect on corn seedling growth]. AB - By using modified PVK plate method, 44 strains of phosphate-solubilizing bacteria (PSB) were isolated from the root surface of wild plants growing well in calcareous soil. After 7-day incubation in a liquid medium NBRIP, the total P in the cultures of strains K3 and K9 reached 643.2 microg x ml(-1) and 608.5 microg x ml(-1), and soluble P was 584.8 microg x ml(-1) and 606.4 microg x ml(-1), respectively. About 12.9% of tricalcium phosphate was dissolved, being 9.5 times higher than CK. Pot experiment showed that the plant height, stem diameter, and dry mass of corn seedling were significantly higher in PSB treatments than in CK. Applying PSB agent with manure as a carrier could significantly increase the seedling's dry mass, in comparing with applying PSB agent alone. The strains K3 and K9 were identified as Pseudomonas. PMID- 17396509 TI - [Effects of planting density and nitrogen application rate on soil microbial activity under wheat/forage rape multiple cropping]. AB - A field experiment was conducted to study the effects of planting density and nitrogen application rate on the topsoil (0-15 cm) microbial activity under wheat/forage rape multiple cropping. The results showed that multiple-cropping forage rape with wheat could significantly increase soil microbial biomass C (Cmic), soil microbial biomass N (Nmic), soil bacteria number (SBN), soil fungi number (SFN) and soil actinomyces number (SAN), but decrease soil microbial biomass C/N (Cmic/Nmic). The Cmic/Nmic and SBN increased with increasing planting density of forage rape, while Nmic and SAN were in adverse. SFN increased significantly with increasing nitrogen application rate, but Cmic and Nmic decreased first, increased then, and decreased again, with the highest in treatment 1000 kg x hm(-2) N. Also with increasing nitrogen application rate, the SFN and SAN during harvest stage of forage rape decreased first and increased then, while the SAN during seedling stage increased first and decreased then. Soil microbial activities at rape harvest stage were all higher than those at seedling stage, except for SAN in treatment 600 kg x hm(-2) N. SBN and SAN were positively correlated with Cmic and Nmic, but negatively correlated with/Nmic. No significant correlation was observed between SFN and Cmic, and SMBN and Cmic/Nmic. PMID- 17396510 TI - [Biodegradation of landfill leachate in soil]. AB - With aerobic and anaerobic incubation tests, this paper studied the biodegradation of three kind landfill leachates in acidic and calcareous soils. The leachates were collected from a landfill just receiving refuse (fresh sample) and the landfills having received refuse for 4-5 years (Tianjingwa sample) and 12 years (Shuige sample). The results showed that in the first seven days of incubation, these three landfill leachates degraded more quickly. Under aerobic condition, the apparent degradation rate of fresh sample, Tianjingwa sample and Shuige sample was 88.9%, 60.5% and 25.0% in acidic soil, and 96.6%, 80.4%, and 65.0% in calcareous soil, respectively. Seven days after, a lower degradation rate was observed. In same test soils, the shorter the landfilling age, the higher apparent degradation rate of the leachates was. Similar results were obtained under anaerobic condition, but the degradation rates were lower. The degradation of test landfill leachates fitted first-order kinetics model well, with a half-life of 12-16 days for fresh sample, and 20-30 days for Tianjingwa and Shuige samples. Once the leachates penetrated into soil, their degradation quickened greatly, suggesting that soil treatment of landfill leachate could have definite efficacy. PMID- 17396511 TI - [Effects of PCBs on Aegiceras corniculatum seedlings growth and membrane protective enzyme system]. AB - With pot experiment, this paper studied the effects of PCBs (0, 180, 900, 1800 and 2700 microg x kg(-1)) on the seedlings growth and the leaves chlorophyll content, MDA content, and membrane protective enzyme system of Aegiceras corniculatum. The results showed that PCBs had definite stimulative effects on the seedlings growth. The stem height, basal stem diameter, and stem volume were all increased with increasing PCBs concentration. Within the test range of PCBs, the chlorophyll content and chlorophyll a/b ratio maintained at a relatively stable and normal level. The decrement of chlorophyll content and the increment of chlorophyll a/b ratio were less than 25% and 10% of the control, respectively. With increasing PCBs concentration, SOD activity increased first and decreased then, while POD activity and MDA content were in adverse. It was suggested that A. corniculatum could endure and adapt definite concentrations of PCBs through physiological and biochemical mechanisms, being available to the phytoremediation of PCBs pollution. PMID- 17396512 TI - [Free radicals in Carassius auratus liver: their generation and oxidative stress induced by 2,4,6-trichlorophenol]. AB - With electron paramagnetic resonance (EPR) technique, this paper studied the free radicals generation and oxidative stress in Carassius auratus liver after 4, 8, 12, 24 and 72 hours of 2, 4,6-trichlorophenol (2,4,6-TCP) intraperitoneal injection. The results showed that 2,4,6-TCP significantly promoted the generation of free radicals in C. auratus liver. The superoxide dismutase (SOD) and glutathione-S-transferase (GST) activities were induced significantly, while catalase (CAT) activity was inhibited, and glutathione (GSH) content had little difference with the control. A significant increase was observed in the contents of oxidized glutathione (GSSG) and malondialdehyde (MDA). PMID- 17396513 TI - [Characteristics of multi-scale temporal-spatial distribution of urban heat island in Guangzhou]. AB - Hourly data were collected from 47 automatic weather stations in Guangzhou to calculate the Guangzhou urban heat island (UHI) intensity in January, April, July and October 2001. The Kriging interpolation showed that the spatial variation of Guangzhou UHI was closely correlated with underlying surface, man-made heat, atmospheric pollutant and weather system, and the analysis on the temporal variation from January 2001 to February 2002 by Morlet wavelet transform revealed that there were multi-scale UHI periods. The main time scales were daily, weekly and bimonthly periods, and the secondary time scale was trimestral period. Sea land breeze, man-made heat, long wave radiation of earth surface, and heat exchanges on earth surface affected the daily period, human activities affected the weekly period, solar radiation and weather system affected the bimonthly, period, while monsoon affected the trimestral period. This temporal-spatial variation of Guangzhou urban heat island was deeply affected by the coactions of weather system, long wave radiation, underlying surface, atmospheric pollutant, man-made heat, and heat exchanges on earth surface, etc. PMID- 17396514 TI - [Spatial correlation of active mounds locative distribution of Solenopsis invicta Buren polygyne populations]. AB - By using geostatistic method, this paper studied the spatial distribution patterns of the active mounds of Solenopsis invicta Buren polygyne populations in Wuchuan and Shenzhen, and built up the spherical models of the interval distances and semivariances of the mounds. The semivariograms were described at the two directions of east-west and south-north, which were obviously positively correlated to the interval distances, revealing that the active mounds in locative area were space-dependent. The ranges of the 5 spherical models constructed for 5 sampling plots in Wuchuan were 9.1 m, 7.6 m, 23.5 m, 7.5 m and 14.5 m, respectively, with an average of 12.4 m. The mounds of any two plots in this range were significantly correlated. There was a randomicity in the spatial distribution of active mounds, and the randomicity index (Nugget/Sill) was 0.7034, 0.9247, 0.4398, 1.1196 and 0.4624, respectively. In Shenzhen, the relationships between the interval distances and semivariances were described by 7 spherical models, and the ranges were 14.5 m, 11.2 m, 10.8 m, 17.6 m, 11.3 m, 9.9 m and 12.8 m, respectively, with an average of 12.6 m. PMID- 17396515 TI - [Secondary productivity of macrobenthos in Jiaozhou Bay]. AB - From September 2003 to September 2004, seven cruises bimonthly survey was conducted on the macrobenthos at five research stations in the Jiaozhou Bay, and the abundance, biomass, secondary productivity, and P/B value of the macrobenthos were studied by using Brey' s empirical formula. The results showed that the secondary productivity of the macrobenthos was averagely 47.34 g x m(-2) x a(-1), and the mean P/B value was 0.58. Correlation analysis indicated that Chl-a was the important environmental factor affecting the secondary productivity. A comparative study on the southern Yellow Sea, Bohai Bay and other areas also proved that on local scale, Chl-a was the important factor affecting the secondary productivity of the benthic community, but on regional scale, water depth was more determinative, i.e., this productivity was decreased with increasing water depth. The greatest contributor to the secondary productivity of macrobenthos in studied areas was Ruditapes philippinarum, the most important economic bivalve in the Jiaozhou Bay, and its population was dominated by the individuals with the ages of less than one year, one year, and two years. PMID- 17396516 TI - [Selective feeding of Calanus sinicus on harmful algal blooms species in East China Sea in spring]. AB - From April 27th to June 5th 2005, the selective feeding of Calanus sinicus on harmful algal blooms (HABs) species was studied by in situ feeding experiments at six typical stations in a high frequency red tide occurring region of East China Sea. The results showed that the selective feeding behavior of C. sinicus was dependent on food density. It had an obvious food selectivity when the abundance of phytoplankton cell was low, but the selectivity changed from preference to discrimination at high phytoplankton cell abundance, with a threshold value of 329 cells ml. The selective feeding of C. sinicus was also dependent on food composition. It showed no selective feeding or discrimination on microzooplankton at most stations, but preferred to P. delicatissima and disliked Karenia mikimotoi at HABs occurring station, which induced the succession of phytoplankton community to K. mikimotoi blooming. PMID- 17396517 TI - [Macrobenthos in Jinping reach of Yalongjiang River and its main tributaries]. AB - In this paper, an investigation was made on the macrobenthos at 25 sites in the Jinping reach of Yalongjiang River and its main tributaries in May and November 2004, aimed to study the relationships between macrobenthos distribution and environmental factors. The results showed that the dominant species of macrobenthos was Baetis sp. (Baetidae, Ephemeroptera), with a relative abundance being 14.9% and 27.0% in May and November, respectively. Based on the species composition and their relative abundance, and by using two-way indictor species analysis (TWINSPAN) and detrended correspondence analysis (DCA), the macrobenthods was divided into four groups, according to the habitat types of the sites. Canonical correspondence analysis (CCA) on the relationships between macrobenthods community structure and environmental factors indicated that in May, elevation and water hardness and NH4+ -N content were the main environmental factors affecting macrobenthods distribution, while in November, elevation, water hardness, NH4+ -N and SiO2 contents were the main ones. PMID- 17396518 TI - [Relationships among planktons DNA sequence diversity, water quality and fish diseases in Siniperca chuatsi ponds]. AB - By using random amplified polymorphic DNA (RAPD) technique, this paper studied the alpha-diversity of plankton communities and its relationships with water quality and fish diseases in 7 Siniperca chuatsi ponds, as well as the effects of stocking density and a new culture model on the diversity and water quality. The results showed that there was a significant negative correlation between the DNA sequence diversity of plankton communities and water quality index, and high stocking density decreased the DNA sequence diversity and increased the water quality index. The new culture model with short culture period, low stocking density and high feeding stuff input had a greater damage on the water environment. Hierarchical cluster analysis indicated that there existed similarities in the DNA sequences of plankton communities and the physicochemical properties of water bodies in the ponds with fish diseases, which provided a possibility to predict the diseases occurrence in Siniperca chuatsi ponds. PMID- 17396519 TI - [Overcompensation effect of Pavlova viridis under ultraviolet (UV-B) stress]. AB - To examine the existence of overcompensation growth of microalgae, this paper studied the effects of UV-B stress on the production of bioactive substances during the overcompensation of Pavlova viridis. Six groups of UV-B treatments 18, 36, 54, 65, 86 and 108 J x m(-2) as well as the control were set up for this study. After the UV-B stress relieved, the treated groups and the control were simultaneously cultured for 12 days under the same inoculation density and normal culture conditions, and the parameters OD, biomass, chlorophyll a, carotenoid, cellular protein and carbohydrate were measured. The results showed that under UV B stress, the growth of P. viridis was inhibited significantly (P < 0.05), with a decrease of relative growth rate being 16.15% - 60.00%, compared with the control. But, after the relief of UV-B stress, the growth rate during anaphase was significantly higher (P < 0.05) in UV-B treatments than in the control, indicating the occurrence of overcompensation growth. On the 12th day of culture, the OD, biomass, chlorophyll a, carotenoid, cellular protein and carbohydrate in treated groups were 22.38%, 15.00%, 26.15%, 23.81%, 11.63% and 27.58% higher than those in the control, respectively. The overcompensation growth of P. viridis provided an effective way to exploit the bioactive substances of microalgae. PMID- 17396520 TI - [Relationships between copepods and Eriocheir sinensis larvae in ecological breeding ponds]. AB - The study on the relationships between two copepod species Eurytemor affinis and Sinocalanus tenellus and Chinese mitten crab Eriocheir sinensis larvae in the ecological breeding ponds of E. sinensis larvae in Liaohe Delta district of Panjing City, Liaoning Province showed that E. affinis and S. tenellus had significant effects on the survival rate of first stage zoae (Z1). The higher the density of the two copepod species, the lower the survival rate of Z, was (P < 0.01). Third stage zoae (Z3) could prey on the copepods nauplii, and the feeding amount was increased with the increasing density of the nauplii (P < 0.01). Fifth stage zoae (Z5) and megalopae could easily prey on E. affinis, but not on S. tenellus. The megalopae yield in the ecological breeding ponds was negatively correlated with the quantity of the copepods during early breeding stage of E. sinensis larvae, and there was a significant difference among the megalopae yields when different quantity of copepods existed (P < 0.01). Some measures for controlling and utilizing the copepods in ecological breeding ponds of E. sinensis larvae were put forward. PMID- 17396521 TI - [Spatial distribution and its environmental analysis of chironomid larvae in Fuxian Lake of Yunnan Province]. AB - A field survey was made on the chironomid larvae in Fuxian Lake of Yunnan Province from June to July 2005. A total of 6 species belonging to 5 genera were collected, among which, Procladius choreus and Chironomous plumosus were the dominant species, with a sum of their relative density and of relative biomass being 94.9% and 97.5%, respectively. The average density of chironomid larvae in the lake was (275 +/- 333) ind x m(-2), and the average biomass was (0.642 +/- 0.763) g x m(-2) x P. choreus distributed all over the lake, C. plumosus mainly distributed in the water area at the south of Mingxingyudong, while other species distributed locally. Micropsectra sp., a typical species distributing in deep oligotrophic water, had an average distribution water depth of 107.2 m. The density of chironomid larvae had a significant positive correlation with water depth, and increased in the sequences of sand or gravel bottom < sand bottom < mud bottom, and submerged macrophyte area < no submerged macrophyte area. Compared with the survey data around 1980, the occurrence frequency and standing stock of chironomid larvae increased greatly, and the indicator species C. plumosus for eutrophic water appeared, indicating that the nutritional level of Fuxian Lake was being improved. PMID- 17396522 TI - [Nitrification-denitrification and N2O emission of typical Calamagrostis angustifolia wetland soils in Sanjiang plain]. AB - With intact soil core and by using acetylene inhibition method, this paper measured the N2O emission and denitrification rates of typical Calamagrostis angustifolia wetland soils in Sanjiang Plain, analyzed their relationships with environmental factors, and estimated the total amounts of N2O emission and denitrification loss. The results showed that meadow marsh soil and humus marsh soil had a similar change range of N2O emission rate (0.020-0.089 kg N x hm(2) x d(-1) and 0.012-0.033 kg N x hm(2) x d(-1), respectively), but the former had a much higher N2O emission rate than the latter, and the difference was significant (P < 0.05). As for denitrification rate, its change range was 0.024-0.127 kg N x hm(2) x d(-1) for meadow marsh soil and 0.021-0.043 kg N x hm(2) x d(-1) for humus marsh soil. Meadow marsh soil also had a higher denitrification rate than humus marsh soil, but the difference was not significant (P > 0.05). In meadow marsh soil, nitrification played an important role in N2O emission and nitrogen loss; while in humus marsh soil, denitrification was the main process inducing N2O emission and nitrogen loss. For these two soils, nitrogenous compounds were not the important factor affecting nitrification-denitrification. In meadow marsh soil, temperature had more evident effect, where nitrification-denitrification had a significant positive correlation with the soil temperature at the depths of 5 cm, 10 cm and 15 cm (P < 0.05). Soil moisture condition was another important factor inducing the difference of N2O emission and denitrification rates. In growth season, the amount of N2O emission and denitrification loss was 5.216 kg N x hm(-2) and 6.166 kg N x hm(-2) for meadow marsh soil, and 3.196 kg N x hm(-2) and 4.407 kg N x hm(-2) for humus marsh soil, respectively. In the denitrification productions of meadow marsh soil and humus marsh soil, the maximum value of N2O/N2 ratio was 5.49 and 3.76, respectively, indicating that the proportion of N2 in denitrification productions was higher in humus marsh soil than in meadow marsh soil, and the seasonal waterlogged condition could induce the decrease of N2O/N2 ratio. PMID- 17396523 TI - [Effects of exogenous metallothionein on thermoresistance and SOD gene expression of dairy cattle]. AB - To approach the effects of exogenous metallothionein (Zn-MT) on the thermoresistance and SOD gene expression of dairy cattle, an experiment was conducted with 28 lactating cows, which were randomly allocated to groups A, B, C and D, and supplemented with 0, 6.0, 12.0 and 16.0 mg Zn-MT x capita(-1), respectively, by intravenous injection. The results showed that the pulse, breath rate, and serum MDA content of the cows in groups B, C and D were lower (P < 0.05 or P < 0.01), while their milk yield, serum- and milk MT contents, blood GSH-PX activity, erythrocyte SOD activity, and SOD gene expression level were higher (P < 0.05 or P < 0.01) than those in group A. All the test indices of the cows in groups C and D were superior (P < 0.05 or P > 0.05) than those in group B, but no significant difference (P > 0.05) was observed between groups C and D. Exogenous Zn-MT had the best effects on the thermoresistance and SOD gene expression of dairy cattle 30 days after injection. All of these suggested that exogenous Zn-MT should be a physiologically active substance effective to the thermoresistance and SOD mRNA expression of dairy cattle, and presented time- and dose-dependent effects. PMID- 17396524 TI - [Research progress on methane oxidation in landfill cover soil]. AB - Methane is the second largest contributor after carbon dioxide to global warming, while landfill is considered as one of the major sources of methane emission, accounting for 1.5%-15% of the global methane sources. Methanotrophic microorganisms play an important role in regulating global methane content, and landfill cover soil is proved to have high capacity of methane oxidation. The study of methanotrophic microorganisms in landfill cover soil and their mechanisms in methane oxidation becomes one of the hot research fields in environmental science and applied microbiology. This review summarized the recent progress on the research of methanotrophic microorganisms, mechanisms and dynamics of microbial methane oxidation, co-metabolism of methane and trace landfill gases, and environmental factors affecting methane oxidation in landfill cover soil. Some perspectives for further research on methanotrophic microorganisms in landfill cover soil were discussed. PMID- 17396525 TI - [Research advances in diet selection mechanisms of grazing herbivores]. AB - Diet selection is a complicated topic. This paper reviewed the multi-disciplinary research advances in the diet selection mechanisms of grazing herbivores. There were two hypotheses about the mechanisms. One assumed that the diet selection behaviors were inherited and innate, while the other supposed that they were from learning, including post-ingestive feedback, trial-and-error learning, social learning, spatial memory, visual cues, and skin-defense. Four hypotheses on diet selection strategies including RT, MVT, OFT and MTD of grazing herbivores were summarized, and the prospects of grazing herbivores diet selection study, such as spatial-scale of diet selection, physiological factors of digestion, and interactions of multiple affecting factors, were discussed. PMID- 17396526 TI - [Methodology and application of soil metagenomics]. AB - Culturable microorganisms provide very limited information on soil microbial diversity, because only 0.1%-10% of soil microorganisms can be cultured under conventional laboratory conditions. A novel culture-independent approach, i.e., metagenomic methodology, has been developed as an effective tool for the discovery of new natural products and microbial functions. Soil metagenomic methods, comprising isolation of soil DNA and construction and screening of clone libraries, enable to peep at more complete scenario of soil microbial communities, and thus, to better understand their interactions. This methodology is of great potential for use in the studies of soil microbial communities and their functional genes, and in the discovery of new biocatalysts for industry and pharmacy. This review summarized some advances of soil metagenomic study. PMID- 17396527 TI - [Effects of aniline and phenol on freshwater algae growth]. AB - By the methods of bioassay, this paper studied the effects of aniline or phenol on the growth of Chlorella pyrenoidosa and Scenedesmus obiquus. The results showed that these two compounds had evident effects on the growth of test algae species. For the same species, aniline was more toxic. Under the same concentration of the compounds, S. obiquus was more sensitive than C. pyrenoidosa. These two algae species could degrade or absorb parts of the compounds, and phenol in particular, when their concentrations were lower. PMID- 17396528 TI - [Characteristics of arthropod community in alpine cabbage fields]. AB - The study on the community structure of arthropod in the alpine cabbage fields of Hubei Province showed that the dominant pests were Brevicoryne brassicae, Mamestra brassicae and Plutella xylostella, while the dominant natural enemies were Diaeretiella rapae, Cotesia plutella, Erigonidum gramiaicolum and Syrphus corollae. The richness, diversity index, evenness index and dominance index of pest and natural enemy sub-communities all changed with time. The dominance index of pest sub-community was higher, while its diversity and evenness indices were lower than those of natural enemy sub-community. Based on fuzzy clustering analysis, the pest and natural enemy subcommunities of 14 time sequences were grouped into 4 and 3 sorts, respectively. PMID- 17396529 TI - [Relationships between eggplant leaf pubescent and its resistance to Polyphagotarsonemus latus]. AB - With 27 eggplant varieties as test objects, this paper studied the effects of their leaf pubescent on the field population density of Polyphagotarsonemus latus, its injury index on eggplant leaf, and population growth rate. The results showed that the density and length of leaf pubescent differed with eggplant varieties. For the same variety, leaf pubescence had a higher density but a shorter length on reverse side than on obverse side. Among the test varieties, 7 resistant varieties had a significantly higher mean pubescence density than 6 susceptive varieties on their reverse side leaf. The higher the pubescence density on reverse side eggplant leaf, the less field population density of P. latus and the lower injury index of eggplant leaf, suggesting a positive correlation between the pubescence density on reverse side eggplant leaf and the resistance of eggplant to P. latus. PMID- 17396530 TI - Routine omissions of care. PMID- 17396531 TI - The challenge of assessing and diagnosing acute abdomen in tetraplegics: a case study. AB - The assessment and diagnosis of an acute abdomen in a spinal cord-injured patient presents a significant challenge to even the most experienced practitioners because of the muted or altered presentations secondary to paralysis. Because the patient with spinal cord injury is at increased risk for peptic or duodenal ulcers and a number of other gastrointestinal emergencies, clinicians need to maintain a high index of suspicion for acute abdomen when working with this population. PMID- 17396532 TI - Postpartum cerebral angiopathy: a case study. AB - Stroke during pregnancy is a leading cause of maternal death, but the risk of stroke does not end with delivery. Postpartum cerebral angiopathy is a rare condition that affects otherwise healthy women who have undergone a normal pregnancy followed by an uncomplicated delivery. It can cause ischemic or hemorrhagic stroke, or both, and usually occurs within the first week following delivery. Although neurological deficits may be minimized if the condition is recognized early, postpartum cerebral angiopathy may be devastating. It is of utmost importance for nurses to recognize the symptoms of postpartum cerebral angiopathy early and be familiar with disease progression and treatment to prevent further neurological compromise. PMID- 17396533 TI - Psychometric testing of the Americanized version of the Guy's Neurological Disability Scale. AB - The Guy's Neurological Disability Scale (GNDS) was originally developed in the United Kingdom to measure disability in multiple sclerosis (MS). The purpose of this study was to test the reliability, validity, and sensitivity of the Americanized version of the GNDS. The following five research questions were considered: (1) Is there convergent validity of the Americanized version of the GNDS as evidenced by a significant relationship with the Short Form 36 (SF-36) and Activities of Daily Living Self-Care for MS Scale? (2) Does the Americanized version of the GNDS have the same four-factor structure as reported in the psychometric testing of the original GNDS? (3) Is the Americanized version of the GNDS a reliable measure when examined with a 2-week retest? (4) Does the Americanized version of the GNDS have internal consistency? (5) Is the Americanized version of the GNDS sensitive to changes in neurological disability from MS over time? Participants were diagnosed with relapsing-remitting MS and a history of one or more relapses in the previous 2 years. The sample included 253 participants--87% were women (n = 219), 13% were men (n = 32), and two participants did not indicate their sex. They ranged in age from 22 to 77 years (M = 46, SD = 9). The Americanized version of the GNDS was found to be a reliable, valid, and sensitive multidimensional measure of neurological disability for individuals with MS. PMID- 17396534 TI - Spinal metastasis in glioblastoma multiforme: a case study. AB - Glioblastoma multiforme (GBM) is the most common primary malignant brain tumor. Spinal metastasis is rare. However, as local control of the primary tumor improves, metastatic disease is increasingly possible. The neuroscience nurse plays an instrumental role in caring for patients with GBM, developing a plan of care based on each patient's needs and outcomes. PMID- 17396535 TI - Normal pressure hydrocephalus. AB - Normal pressure hydrocephalus (NPH) is a neurological disorder characterized by a triad of symptoms: gait disturbance, dementia, and urinary incontinence. To date, the treatment of choice is cerebrospinal fluid diversion with a shunt. Because the clinical presentation of NPH mimics other neurological disorders, or can be perceived by healthcare providers as a natural aging process, it can be misdiagnosed or go undetected for many years. Patients with NPH suffer from debilitating physiological impairments as well as psychological disturbances. There are many different healthcare concerns in this population, including quality-of-life and safety issues. To achieve a more expedient diagnosis and treatment plan, nurses must be educated about this disorder. PMID- 17396536 TI - Nursing issues in caring for children with moyamoya vasculopathy and their families. AB - Moyamoya vasculopathy is a rare, progressive neurovascular condition that may cause recurrent transient ischemic attacks, ischemic strokes, hemorrhagic strokes, or neurologic decline in children. Children with moyamoya disease have no clear contributing etiology, but children with moyamoya syndrome have contributory diagnoses such as Down syndrome or neurofibromatosis. The concerns and lived experiences of children affected by this disease and their families have not been well explored or addressed in the nursing literature. Nurses who understand this lived experience increase their own knowledge of the disease so they can manage the complex medical issues, educate families about the disease, and provide emotional support. PMID- 17396537 TI - Recognizing thyroid storm in the neurologically impaired patient. AB - Thyroid storm is a life-threatening complication of thyroid dysfunction that is manifested by signs of cardiac arrhythmias, fever, and neurological impairment. These symptoms can easily be attributed to a multitude of factors commonly seen in neurological intensive care units, making the recognition and diagnosis of this event difficult. In this case study, a patient presents with a complicated course of hospitalization exacerbated by thyroid storm. Early nursing care and medical collaboration offset a potentially fatal condition. PMID- 17396538 TI - Mild traumatic brain injury: an update for advanced practice nurses. AB - Nearly 75% of persons with brain injury experience a mild injury. These people do not often enter the healthcare system by traditional means, nor do they always present with visible signs and symptoms of injury. In fact, people who experienced brain trauma are likely to seek help in primary care settings and from advanced practice nurses (APNs). Because the symptom experience can be complicated by impaired perception or mood, delays in seeking help, and faulty explanations for their symptoms, APNs need to rule out competing diagnoses, offer brief psychoeducational treatment, and refer the person to an appropriate specialist for therapy when needed. PMID- 17396539 TI - Pharmacological management of secreting pituitary tumors. AB - Because pituitary adenomas can present in many ways, nurses need to be aware of the signs and symptoms of different hormone-secreting tumors and their related pharmacologic treatment. Although long-term medical management of secreting tumors and their hormonal complications is usually carried out on an outpatient basis, diagnosis often occurs during inpatient care. Consequently, nurses in both settings need to be knowledgeable about medications used, outcomes expected, and adverse effects experienced. Patient care should be centered around education and management of complications as patient compliance is increased with knowledge. PMID- 17396540 TI - Predictive value of biochemical markers in stroke. AB - Biochemical markers of cerebral ischemia and subsequent inflammation represent a potential diagnostic and predictive modality in acute ischemic stroke. Measurement of these markers may improve clinical functions ranging from diagnosis of ischemic stroke to prediction of functional outcome at 3 months. Predictive models, using various biochemical markers, continue to be investigated. PMID- 17396541 TI - Uninsured Arkansas. PMID- 17396542 TI - Defendants, hospital and physician below the standard. PMID- 17396543 TI - Access to care: Medicaid recipients have access to a medical home. PMID- 17396544 TI - RQI grant offers chance to enhance state's health system. CHCS, RWJ funding support effort by Medicaid, AFMC. PMID- 17396545 TI - Minimally invasive joint replacement. PMID- 17396546 TI - Charcot arthropathy. An often overlooked complication of diabetes mellitus. AB - Charcot's arthropathy is a rare complication of diabetes that can result in significant morbidity. It may have a similar presentation to cellulitis, which often delays its recognition. While the pathogenesis remains unclear, timely diagnosis and treatment can help prevent the deformities associated with the late stages. It is important for physicians to recognize this complication early and use a team approach that includes an experienced orthopedic surgeon and/or podiatrist. We present a case and review the presentation, diagnosis and treatment of Charcot's arthropathy. PMID- 17396547 TI - Advancing the science of nursing education: rethinking the meaning and significance of impact factors. AB - It is not the case that impact factors are unhelpful and that their use should be abolished. Indeed, ISI is continuing to refine existing and develop new databases to provide important information to researchers, administrators, librarians, and editors. Impact factor data do provide useful information for the review process if used judiciously and with an awareness of what these data do and do not indicate. Perhaps it is timely for members of the nursing discipline to think more broadly about the nature of impact and to talk more about the ways in which review processes can account for the many ways the impact of research can be demonstrated. For example, as part of the review process, reviewers might ask each researcher to provide exemplars from teaching or clinical practice settings in which their research is actually being used. Researchers might also be asked to describe the impact that has occurred in ways other than through publication (e.g., presentations, consulting) and how this impact was determined. Creating review processes that allow researchers to describe their decision making related to disseminating their research and how this reflects their ability to influence the discipline may provide a more realistic picture of impact than calculated figures alone. PMID- 17396548 TI - Learning across the professions: the clergy, a case in point. AB - In this article, we examine similarities and differences in the educational goals and pedagogies of nursing and clergy students. We argue that nurse educators can learn a broader perspective of rationality than the narrow, technical rationality of instrumental problem solving taught in most professional schools. An interpretive form of rationality is needed to address suffering and human concerns in the world. The pedagogies of interpretation, formation, contextualization, and performance used in clergy education offer a more interpretive, historical, and contextual approach to reasoning. PMID- 17396549 TI - Using portfolio evaluation for program outcome assessment. AB - Portfolio evaluation is an efficient and effective strategy to objectively demonstrate the accomplishment of program objectives using students' perceptions of their academic experiences. A portfolio evaluation tool was developed to quantitatively measure students' perceptions of their achievement of the nursing program's goals and objectives. Data were collected from a convenience sample of 26 portfolio evaluations. Reliability testing showed the tool has high interrater reliability, with Cohen's Kappa >0.80 for most items. Individual achievement of program goals and objectives ranged from 53.6% to 100%. These data will be used in the next formative evaluation of the curriculum to revise program objectives and correspondneing course objectives and assignments.ty PMID- 17396550 TI - Undergraduate nursing students' knowledge of and attitudes toward aging: comparison of context-based learning and a traditional program. AB - The purpose of this study was to investigate nursing students' knowledge of and attitudes toward older adults in the first and fourth years of a baccalaureate program, following the introduction of a context-based learning (CBL) curriculum, and to compare the fourth-year CBL student findings to those of fourth-year students in the final year of the traditional, lecture-based baccalaureate program. The Facts on Aging Questionnaire was used to assess knowledge, and the Aging Semantic Differential was used to assess attitudes toward aging related to societal influences. Although there were differences in knowledge and attitudes between fourth-year CBL and fourth-year traditional students, the differences were not significant. These findings support earlier work that an integrated curriculum may not significantly improve knowledge of age-related changes nor positively influence attitudes that are already positive. The Reactions to Ageing Questionnaire was used to examine students' attitudes toward personal aging. There was a significant positive increase in CBL students' attitudes toward personal aging from the first to fourth years of the program. This suggests that CBL learning fosters an inner maturity toward personal aging. PMID- 17396551 TI - Critical assumptions: thinking critically about critical thinking. AB - The concept of critical thinking has been featured in nursing literature for the past 20 years. It has been described but not defined by both the American Association of Colleges of Nursing and the National League for Nursing, although their corresponding accreditation bodies require that critical thinking be included in nursing curricula. In addition, there is no reliable or valid measurement tool for critical thinking ability in nursing. As a result, there is a lack of research support for the assumptions that critical thinking can be learned and that critical thinking ability improves clinical competence. Brookfield suggested that commitments should be made only after a period of critically reflective analysis, during which the congruence between perceptions and reality are examined. In an evidence-based practice profession, we, as nurse educators, need to ask ourselves how we can defend our assumptions that critical thinking can be learned and that critical thinking improves the quality of nursing practice, especially when there is virtually no consensus on a definition. PMID- 17396552 TI - Medication error reduction and the use of PDA technology. AB - The purpose of this study was to determine whether nursing medication errors could be reduced and nursing care provided more efficiently using personal digital assistant (PDA) technology. The sample for this study consisted of junior and senior undergraduate baccalaureate nursing students. By self-selection of owning a PDA or not, students were placed in the PDA (experimental) group or the textbook (control) group, provided with a case study to read, and asked to answer six questions (i.e., three medication administration calculations and three clinical decisions based on medication administration). The analysis of collected data, calculated using a t test, revealed that the PDA group answered the six questions with greater accuracy and speed than did the textbook group. PMID- 17396553 TI - Making a difference with combined community assessment and change projects. AB - Nursing students value projects that enable them to make a difference in the lives of others. Two major group projects, assessment of an identified community and completion of a leadership change project, required during their last semester were combined to provide a meaningful experience for students. In addition to discussing ways to combine these projects, this article shares methods to build team cohesiveness. PMID- 17396554 TI - Evaluation of undergraduate students using Objective Structured Clinical Evaluation. AB - This study proposed to develop an Objective Structured Clinical Evaluation (OSCE) for senior undergraduate nursing students. This evaluation process uses a simulated and standardized format to measure synthesis of knowledge and clinical skills. The benefit of OSCE is that it provides a formative evaluation for both students and the educational institute. The study included developing case studies, identifying or developing assessment tools, hiring and training standardized patients, obtaining a testing site, and analyzing data. A total of 49 students were assessed on three of six patient simulation stations and one of two nonpatient stationary stations. Both formative and summative evalu- PMID- 17396555 TI - Evidence-based strategies of graduate students to achieve success in a hybrid Web based course. AB - Web-based hybrid courses are gaining in popularity in institutions of higher learning for both undergraduate and graduate nursing education. The purpose of this study was to examine how predictive the five self-regulatory resource management strategies of time management, study environment, effort regulation, help seeking, and peer learning are in determining whether a student will be successful academically within a hybrid learning environment. The sample consisted of 38 graduate nursing students enrolled in two sections--one hybrid and the other lecture--of a health care economics course at a major, public, urban, 4-year university. The results of the study revealed that students in the hybrid section had significantly higher end-of-course grades and a significantly higher favorable rating (affective behavior) of their method of instruction. Of the five resource management strategies examined, only help seeking showed a significant correlation with end-of-course grades in both sections. PMID- 17396556 TI - Experiencing the effect of teaching and learning styles on skill mastery. PMID- 17396557 TI - [The regulatory role of mitochondrial dysfunction in hypoxia and its interaction with transcription activity]. AB - The mitochondrial respiratory chain participates in the performance of the signal system, which activates the realization of metabolic compensatory processes and coupled functional response to both single and repeated, long-term exposure to acute hypoxia. Under the conditions of reduced oxygen delivery to cells the mitochondrial respiratory chain is involved in the process of oxygen homeostasis regulation and modulates oxygen consumption, the rate of oxygen delivery from the extracellular milieu to mitochondria, and energy synthesis, activating hypoxia specific transcription factors as well. PMID- 17396558 TI - [Properties of NAD-dependent brain enzymes under the conditions of hypoxia and ischemia]. AB - The catalytic properties of mitochondrial and cytoplasmic isoenzymes of NAD dependent brain malate dehydrogenase (MDH) were studied under hypoxic or ischemic conditions. Hypoxia was modeled in animals in pressure chamber, while ischemia was achieved via bilateral ligation of common carotic arteries. The properties of MDH in mitochondria of rat brain were studied; they were significantly different from those of MDH purified from bovine brain. The study revealed the importance of mitochondrial membranes for the regulation of malate dehydrogenase catalytic properties in brain mitochondria. Cerebral ischemia changes mitochondrial malate dehydrogenase significantly, which demonstrates disorder in MDH-membrane interaction. Cytoplasmic enzyme displays high activity and stability of its catalytic properties. Under cerebral hypoxia or ischemia catalytic properties of cytoplasmic malate dehydrogenase change only slightly, maintaining enzyme activity at a constantly high level. PMID- 17396559 TI - [Dynamic changes in transcription factor HIF-1alpha, rapid response protein, and membrane structure resistance following acute hypoxia]. AB - Within the last decade, a great number of reports have discussed cellular redox signalization depending on the levels of oxygen and reactive oxygen species (ROS). Experiments have proven that ROS can not only be damaging, but are also able to induce the synthesis of cell defense systems. The initiation of redox signal system results in the induction of various transcription factors which response to hypoxia and hyperoxia, an increase in ROS, oxidants etc. The most significant of them is HIF-1alpha, transcription factor playing an important role in the regulation of oxygen homeostasis in the cell as well as in resistance of the heart and the brain to ischemic and reperfusion injury. About 60 genes activated by HIF-1 are known today; among these are genes that code defense systems: cellular antioxidant system, peroxiredoxines, prostaglandins, heme oxygenase, and heat shock proteins. However, despite numerous data on HIF-lalpha expression stimulation or suppression in exposure to ischemia or hypoxia, they are rather contradictory. In this study, changes in HIF-la induction three, six, and twelve hours after acute hypoxia (8% O2 during one hour) were evaluated, and the dynamics of HIF-1alpha level following hypoxia was compared with the dynamics of the levels of rapid response protein, such as inducible heme oxygenase form, HSP70 stress inducible protein, and antioxidant defense enzymes. The findings indicate a nonlinear dynamics of changes in the levels of transcription factors and rapid response proteins with protective function, tissue specificity of their induction, a direct correlation between HIF-1alpha and superoxide dismutase levels in the heart and HIF-1alpha and HSP70 levels in the liver. The stability of membrane structures of different organs and cardiac sarcoplasmatic reticulum Ca pump are maintained by activation of redox signalization and compensatory synthesis of defense proteins. PMID- 17396560 TI - [Protective and damaging effects of periodic cerebral hypoxia: the role of nitric oxide]. AB - Low oxygen delivery to organs and tissues is one of the most life-threatening situations. Periodic hypoxic episodes may have not only damaging, but also protective effects on the organism depending on how long and intensive this factor is. In both cases an important role is played by changes in the synthesis and metabolism of NO. The direction of NO synthesis and, finally, the direction of periodic hypoxia effect is determined by the regimen of hypoxic impact. The effect of NO depends on its concentration. Both NO excess and deficit are very unfavorable to the organism. Sleep apnea syndrome and pulmonary hypertension are typical examples of NO-dependent damaging effects of periodical hypoxia. NO dependent protective effects of adaptation to periodic hypoxia are underlied by moderate stimulation of NO synthesis, which provides both compensation for NO deficit and the limitation of its hyperproduction. In turn, NO may increase the expression of other protective factors, which makes adaptive protection more reliable and durable. Understanding the mechanisms of adaptation to hypoxia will help develop new approaches to the prevention of hypoxia and ischemic lesions and the improvement of adaptive abilities of the organism. PMID- 17396561 TI - [Mitochondrial ATP-dependent potassium channel. 1. The structure of the channel, the mechanisms of its functioning and regulation]. AB - The modern conceptions of the structural organization and functional mechanisms of mitoKATP are discussed in the present review. Methods of the identification of the channel in the inner mitochondrial membrane and in intact cells are considered. The structure and properties of the channel, the mechanisms of its regulation with pharmacological modulators, redox agents (SH agents, NO and ROS), hormones etc. are reviewed. The authors adduce their own data concerning the structural organization, regulation, properties and the functional role of mitoKATP in the cell. PMID- 17396562 TI - [Mitochondrial ATP-dependent potassium channel. 2. The role of the channel in protection of the heart against ischemia]. AB - The cardioprotective properties of pharmacological and metabolic activators of mitoKATP are reviewed. Metabolic activators of the channels and data on their cardioprotective properties are discussed in the review. The authors adduce their own data concerning cardioprotective properties of mitoKATP channel metabolic activator (UDP). In experimental animals, UDP precursors, uridine and UMP, decrease myocardial ischemic alteration index and T-wave amplitude within 60 min after occlusion of the left coronary artery. Both effects are prevented by mitoKATP channel inhibitors, glibenclamide and 5-HD. UMP and uridine possess antiarrhythmic properties as well. These preparations decrease the number of premature ventricular beats, the duration of ventricular tachycardia and fibrillation, and these effects are eliminated mainly by glibenclamide. Thus, mitoKATP plays a significant role in prevention of both ischemic lesions and rhythm disorders. The prospects of application of metabolic activators to prevent and treat myocardial infarction are discussed. PMID- 17396563 TI - [The effects of adaptation to hypoxia on the resistance to neurodegenerative disorders in the brain of rats of different genetic strains]. AB - The aim of the study was to compare the protective effects of adaptation to altitude hypoxia (AH) on neurodegenerative brain disorders (NBD) induced with infusion of beta-amyloid peptides (Abeta) into the brain (imitation of Alzheimer's disease) of rats belonging to two species: Wistar rats (WR) and August rats (AR). Previously it was shown by the authors that WR were less resistant to memory function impairment and open-field activities, induced with Abeta infusion compared with AR. This study showed that preliminary AH significantly restricted brain function impairment induced by Abeta in WR, so AH demonstrated the protective effect in WR. In contrast, in AR preliminary AH provoked those impairments induced by Abeta. The AH protective effect in WR was associated with activation of stress-limiting systems (antioxidant system, NO system). Lack of AH protective effect in AR was associated with lack of activation of these systems in these rats. Thus, the different AH effects on NBD development in WR and AR are obviously determined by hereditary peculiarities of stress-limiting systems in WR and AR. PMID- 17396564 TI - [Energotropic, antihypoxic, and antioxidative effects of flavonoids]. AB - Phytogenous flavonoid-containing agents (PFCA) are able to initiate electron flow bypassing the NAD-dependent region of respiratory chain, which is related with the activity of DT-diaphorase catalyzing two-electron reduction of quinones to hydroquinones and hydrogen peroxide in the presence of NADH and oxygen. This property is dramatically potentiated under the conditions of suppressed electron transport function of mitochondrial enzyme complex I (MEC I). In this process, part of the flow goes to the cytochrome region of respiratory chain and provides recovery of the MEC II and MEC III coupling function. The other part forms a flow of free oxidation which can perform as an additional mechanism normalizing the cell redox potential and aimed at decreasing intracellular acidosis under the conditions of MEC I bypassing. The energotropic effect of PFCA under the conditions of blocked MEC I is best evident at low PFCA concentrations. The ratio of coupled to free oxidation in the presence of PFCA depends on PFCA concentration. At low PFCA concentrations and oxidation of NAD-dependent substrates, both pathways become potentiated to an approximately similar extent, although the coupled oxidation pathway is generally activated earlier. At high PFCA doses, the increase in free oxidation pathway predominates and may result in toxic side effects. PMID- 17396565 TI - [Anatomy education at the beginning of Japan's Meiji era and an anatomical model with topographical expressions of anatomy under the fasciae]. AB - One anatomical model of a human is preserved at Kyushu University. We presume this model to have been made at the beginning of Japan's Meiji era as a copy of the Anatomie clastique, which was designed and manufactured by Louis Thomas Jerome Auzoux and imported from France at the end of the Edo era. The model has labels of anatomical nomenclature in Japanese, which are taken from Kazuyoshi Taguchi's Kaibou-Ranyo, (see text) and the unique expressions of topographical anatomy under the fasciae. These are unique characteristics that cannot be seen on the Auzoux original. This model is supposed to have been made for use in professional medical training, and these characteristics reflect the historical background of the times when Western medicine was being introduced into Japan. At that time, Japan urgently needed training for doctors who were acquiring the techniques of surgery, but there was no system to supply a plentiful number of cadavers for use in anatomical education. PMID- 17396566 TI - [Beginning of modern anatomy in Japan from the perspective of anatomical bibliographies of the Meiji era]. AB - In contrast to the days of the Shogunate during the Edo period when knowledge and information from abroad were derived solely from The Netherlands (so-called Western learning), the establishment of the new Meiji government marked the arrival of a wealth of new knowledge from English and German speaking countries to Japan. As with other academic fields, the field of medicine, particularly anatomy, experienced an influx of many foreign books during this period. In the early to middle Meiji period (1868-1887), anatomy books from English-speaking countries became mainstream, and translations of these books were subsequently published. However, following Japan's decision in the third year of the Meiji era (1870) to model itself after German medicine, and the subsequent introduction of German teachers in Japanese medical education, medicine from English-speaking countries was gradually replaced by German medicine. Consequently, a multitude of German anatomy textbooks began to be imported into Japan during the middle Meiji period. In the later half of Meiji period (1888-1912), sequential publication of books written by Japanese anatomists based on German anatomy books became more common, along with the development of medical personnel who had been taught by foreign teachers. Most of the anatomy textbooks written by Japanese anatomists followed a format based on that of German anatomy textbooks of the time. This format style became well established by the late the end of Meiji period, and continued to be used in subsequently published anatomy textbooks until around the end of World War II. Here we introduce books, such as the translated anatomy books and textbooks, that were published during the turbulent Meiji era, and describe these within a bibliographical context. PMID- 17396567 TI - [Anatomical education in the late Meiji era --Lu Xun, doctor Fujino and their comtemporaries]. AB - Lu Xun studied medicine at Sendai Medical School for 1 and a half years and then changed his course to Literature. In his novel "Doctor Fujino", Lu Xun told his memory on the anatomical notes in which Professor Fujino made numerous corrections. I analyzed the anatomical notes by Lu Xun and his classmates, and revealed the situation of lectures at that time. The teachers drew many anatomical illustrations on the black board with colored chalks. The lecture notes of students may be either clean copies rewritten after lectures or crude notes written during the lectures. When making clean copies, they copied anatomical illustrations in the anatomical textbooks at hand. The anatomical textbooks by Gegenbaur, Rauber and Ishikawa were utilized. Lu Xun made clean copies in the first two months after matriculation, and made crude notes after then. Corrections by Professor Fujino were found in the crude notes for his lectures. PMID- 17396568 TI - [History of collecting cadavers in Japan]. AB - This study investigated how and from where medical students had acquired cadavers for research throughout Japanese history. At the beginning of dissection in the mid Edo era, they cut up executed prisoners granted by the Tokugawa Shyogunate to study internal body parts. After the Meiji Restoration, the social mechanism of delivering cadavers underwent a complete transformation and they began to utilize 1) dead bodies of inpatients who had received free medical treatment and 2) unclaimed bodies mainly from homes for the aged and prisons. It was quite recently that "kentai", voluntary body donation, became common practice of collecting cadavers. Consequently the history of cadavers submitted to dissection faithfully reflects the relation between medical science and society. PMID- 17396569 TI - [Premise of patient safety]. PMID- 17396570 TI - [Current status and issues in hematopoietic stem cell transplantation]. PMID- 17396571 TI - Measuring job stress among hospital nurses: an attempt to identify biological markers. AB - The purpose of this study was to identify biological markers corresponding to job stress among hospital nurses. The subjects of this study were 128 nurses working at a university hospital. The NIOSH job stress questionnaire and the Miki Nurse Stressor 35-item Scale measured their job stress levels. The GHQ28 was also used to measure the subjects' general mental health status. Blood analyses for neuroendocrine function and immunity reaction were performed in order to identify biological markers of job stress. Stress is related to the plasma levels of catecholamine, cortisol, adrenocorticotrophic hormone, and natural killer cell activity, therefore these factors were measured accordingly. In consideration to circadian rhythms, blood was collected from the subjects prior to the start of the day shift. The nurses filled out the questionnaires on the day of the blood tests. In order to investigate the correlation between job stress reactions indicated by the questionnaires and the results of the blood tests, we utilized Pearson's correlation coefficient and partial correlation coefficient for which other affected items were controlled. In this study, significant correlations were found between job stress and biological factors; however, the correlations were not strong. Thus, it can be said that the biological markers associated with a specific kind of job stress remain unclear. In the future, rather than implementing a simple cross-sectional study, a longitudinal study including follow-up research will be more effective in establishing biological markers for job stress. PMID- 17396572 TI - [Maternal filicide in Japan: analyses of 96 cases and future directions for prevention]. AB - OBJECTIVE: Maternal filicide is not an isolated phenomenon. When a mother kills her child, she may be affected by many factors and confronted with different problems based on the child's developmental stage. In this study in Japan, a judicial sample of 96 adult women, convicted in their first trial for the murder or attempted murder of their children, was divided into four groups of mothers according to the age of the victim (25 women killed neonates, 22 women infants, 27 women preschool children, and 22 women schoolchildren and/or teenagers) in order to identify the factors that have a major impact on filicide in each group. METHODS: The socio-demographic, clinical, forensic, circumstantial, and offense characteristics, and legal disposition of 96 cases drawn from judicial records were compared among the four groups using the Kruskal-Wallis test; comparison of two groups was conducted using the Mann-Whitney test. RESULTS: Neonaticide cases were distinguished from the other three groups by marked differences: a significantly higher rate of unmarried mothers, financial difficulties, absence of mental illness, and admission of not wanting an illegitimate child. In the other groups, mental disorders were frequent; in particular, post-partum depression was the primary cause of infanticide. For the two groups of cases involving a child older than one year, filicidal mothers were more affected by circumstantial factors such as health problems of the child or severe marital discord. These problems may then have caused a reactive mental disorder among these mothers. The risk of fatal abuse or neglect was higher for handicapped preschool children. Filicide-suicide was most frequently seen among school-aged children and/or teenagers who had serious behavioral problems, and these children often had a mental disorder. CONCLUSIONS: The classification of maternal filicide by age of the child demonstrated that there are specific issues for each group. Based on these findings, future directions for prevention include: appropriate sex education for youths to avoid unwanted pregnancy; organization of specialized mental health services for mothers with post-partum mental disorder; careful psychiatric risk assessment of mentally ill mothers; and development of diversified social support measures for child-bearing parents, especially those with identifiable financial or social difficulties. PMID- 17396573 TI - The mechanism of action of the natural bioregulator stifun. PMID- 17396574 TI - Study of the mechanism of reversible inhibition of various cholinesterases by thionephosphonates on the basis of data on comparative inhibitory specificity. PMID- 17396575 TI - A study of recombinant human lactoferrin secreted in milk of transgenic mice. PMID- 17396576 TI - Inhibition of migration of monocytes and granulocytes in vivo by the peptide corresponding to sequence 65-76 of monocyte chemotactic protein-1 (MCP-1). PMID- 17396577 TI - Modeling the growth and morphogenesis of malignant tumors. PMID- 17396578 TI - Fine stratified distribution of Gammarus lacustris Sars (Crustacea: Amphipoda) in the pelagic zone of the meromictic Lake Shira (Khakassia, Russia). PMID- 17396579 TI - Construction of chimeric lectins with new sugar-binding properties. PMID- 17396580 TI - Immunological detection of plant protein At-4/1 capable of interaction with viral movement proteins. PMID- 17396581 TI - Apoptotic shrinkage of lymphoid cells: a model of changes in Ion flux balance. PMID- 17396582 TI - Analysis of correlations in location of hydrophobic and hydrophilic monomers in protein sequences. PMID- 17396583 TI - Interaction with DNA as a cytotoxicity factor of a novel glycoside derivative of indolocarbazole. PMID- 17396584 TI - Effect of nitric oxide and hyperthermia on the content of heat shock proteins in tumor cell culture. PMID- 17396585 TI - Identification and determination of glycosides in tobacco leaves by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - HPLC-atmospheric pressure chemical ionization MS (HPLC-APCI-MS) was used to screen and identify glycosides in tobacco leaf. MS/MS and MS3 and photodiode array (PDA) detection were also used in the characterization. A total of 12 glycosides were found and four of them were identified based on their abundant [M + H]+ ions, UV spectra, and MS/MS analysis and they are scopolin, rutin, quercetin-3-glycoside, and kaempferol-3-rutinoside. Analytical characteristics of the method were investigated. The contents of these glycosides were obtained and compared based on the relative peak area to the internal standard in seven kinds of tobacco leaf. PMID- 17396586 TI - Pharmaceutical determination in surface and wastewaters using high-performance liquid chromatography-(electrospray)-mass spectrometry. AB - A method was developed to determine 11 pharmaceutical compounds in water samples. The method uses SPE and HPLC coupled to MS (LC/MS) using ESI in both positive and negative modes. Three different sorbents were compared for the extraction of analytes from river and sewage treatment plant (STP) waters and OASIS HLB provided the best results. For the solid-phase extraction of 500 mL of river water samples, the recoveries were between 41 and 101% with the exception of acetaminophen, salicylic acid and naproxen. The LODs were between 3 and 5 ng/L for all the compounds, except naproxen which had an LOD of 15 ng/L. Acetaminophen, caffeine, carbamazepine, bezafibrate and ibuprofen were found in three of the tested river samples at ng/L levels and among them, the highest values were for caffeine and bezafibrate with 305 and 363 ng/L, respectively. For the influent and effluent water samples of the STP, volumes of 100 and 250 mL were used, respectively, to obtain acceptable recoveries. All the compounds showed recoveries between 33 and 91% for effluent samples and 33-72% for influent samples, with the exception of acetaminophen, salicylic acid and bezafibrate, which had lower recoveries. The method developed enabled pharmaceuticals in the influent and effluent sewage waters to be determined in five campaigns carried out between February 2004 and June 2005. Several pharmaceuticals were found in the influent samples: for instance, maximum concentrations of ibuprofen and caffeine were 6 and 40 microg/L, respectively. PMID- 17396587 TI - Rapid determination of polyprenylated xanthones in gamboge resin of Garcinia hanburyi by HPLC. AB - A rapid ion-pair HPLC method was developed and validated for the determination of eight polyprenylated xanthones including three pairs of epimers, namely morellic acid (MA), 30-hydroxygambogic acid (HGA), 30-hydroxyepigambogic acid (HEGA), isogambogic acid (IGA), epiisogambogic acid (EIGA), gambogenic acid (GNA), gambogic acid (GA), and epigambogic acid (EGA), in gamboge resin of Garcinia hanburyi. The separation was performed on a narrow bore C8 column with isocratic elution using a mixture of methanol-ACN-40 mM KH2PO4 buffer (37.5:37.5:25 v/v/v, containing 0.1% tetradecyltrimethylammonium bromide). The newly developed method was used to determine the contents of the eight compounds present in the gamboge. Results showed that GA and EGA are the dominant components of gamboge. The content ratio of each epimer pair remained constant, indicating that the content ratio of epimers can be used as a specific characteristic for the quality control of gamboge. PMID- 17396588 TI - Enantiomeric separation of chiral pesticides by high performance liquid chromatography on cellulose tris-3,5-dimethyl carbamate stationary phase under reversed phase conditions. AB - Twenty chiral pesticides were tested, of which seven samples were directly separated by HPLC using cellulose tris-3,5-dimethyl carbamate (CDMPC) chiral stationary phase under RP conditions. The influence of mobile phase composition and column temperatures from 0 degrees C to 40 degrees C on the separations were investigated. The mobile phases were methanol/water or ACN/water at a flow rate of 0.8 mL/min with UV detection at 230 or 210 nm. Epoxiconazole, terallethrin, benalaxyl, and diclofopmethyl were observed to obtain the baseline separation under suitable conditions and other pesticides pyriproxyfen, lactofen, and quizalofop-ethyl were separated partially. The retention factors (k) and selectivity factor (alpha) for the enantiomers of most investigated pesticides decreased upon increasing the temperature except for the selectivity factors (alpha) of pyriproxyfen in methanol/water. The ln alpha - 1/T plots for racemic chiral pesticides were linear at the range of 0-40 except for that of pyriproxyfen enantiomers in methanol/water and the chiral separations were controlled by enthalpy. Better separations were not always at low temperature. The elution orders of the eluting enantiomers were determined by a circular dichroism (CD) detector. PMID- 17396589 TI - An alternative multiple-trapping LC-SPE-NMR system. AB - In this paper, we describe approaches that make RP LC-SPE-NMR simpler, and in our opinion, result in more reliable methods for trapping and subsequent transfer of separated trace-level compounds to the NMR. An SPE unit based on a commercially available, low dead-volume 10 port high-pressure column selector gives the possibility of trapping compounds on nine individual SPEs that have standard fittings. This allows the operator to employ specific stationary phases that are not available as SPEs in commercially available LC-SPE-NMR systems. Multiple trappings of small compounds like monuron, 1-(4-chlorophenyl)-3-methylurea, and 4 chlorophenylurea were easily performed employing a porous-carbon SPE material. The system was optimized to elute the SPE-trapped compounds to the NMR probes in as small a volume as possible using back-flushing. The proper match of NMR probe volume and SPE column inner diameter and elution volume was discussed, as well as the necessity of drying loaded SPEs prior to NMR transfer when using porous carbon SPE material. PMID- 17396590 TI - Identification of dimer impurities in ampicillin and amoxicillin by capillary LC and tandem mass spectrometry. AB - A micro-scale liquid chromatography electrospray ionization tandem mass spectrometric method was developed for the identification of polymerized impurities in ampicillin and amoxicillin in aqueous solution. Ampicillin and amoxicillin are broad-spectrum antibiotics and widely used for the treatment of human and animal infections. In this study ampicillin, amoxicillin, and their dimers were trapped in a 5-cm capillary column containing C18 sorbents. The analytes were separated on a reversed-phase column and introduced into the mass spectrometer via a nanospray ion source. An isocratic mobile phase consisting of 1% formic acid-acetonitrile (50:50, v/v) was used. For identification, the fragment ions of the analytes were monitored. The aim of the present study was to develop an optimized quality control method for the analysis of high molecular weight impurities of ampicillin and amoxicillin. PMID- 17396591 TI - Chiral separation of beta-blocker drug nadolol by HPLC--a kinetic and equilibrium study. AB - A Chiralpak AD-H column packed with amylose tris(3,5-dimethylphenylcarbamate) coated on silica gel was used to study the enantioseparation of nadolol by HPLC. The bed voidage, axial dispersion coefficient, overall mass transfer coefficients as well as equilibrium constants for the chromatographic enantiomeric separation were evaluated by moment analysis on the basis of the solid film linear driving force model. The equilibrium constants were found to be 3.81, 5.24, 9.45 and 19.41 for the stereoisomers (SRS)-, (SRR)-, (RSS)- and (RSR)-nadolol, respectively. Their overall mass transfer coefficients were found to be 1841.8, 1254.8, 799.4 and 401.7 min(-1) respectively. Temperature effect on the enantiomeric separation and thermodynamic properties including enthalpy and entropy change of binding to the amylose tris(3,5-dimethylphenylcarbamate) stationary phase were also investigated. The moment analysis and the parameters obtained were used to simulate nadolol elution profiles. The simulated results matched the experimental profiles well, which confirmed the validity of model parameters obtained in this study. PMID- 17396592 TI - Enantiomeric resolution of new triazole compounds by high-performance liquid chromatography. AB - Cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) was synthesized and coated on aminopropylsilica to prepare a chiral stationary phase (CSP). HPLC methods were developed for the direct enantioseparation of 12 chiral triazole compounds on the CSP. The separations were made using normal phase methodology with a mobile phase consisting of n-hexane-alcohol (ethanol, 1-propanol, 1-butanol, 2-propanol, and t butanol) in various portions. The column temperatures were studied for the optimization of the resolutions. The effects of structural features of the solutes on the discrimination between the enantiomers were examined. Baseline separation was easily obtained in many cases. PMID- 17396594 TI - Palladium(II) chelate of tetradentate Schiff base as mixed stationary phase for gas chromatography. AB - Bis(acetylacetone)ethylenediiminepalladium(II) (PdAA2en) was examined as a mixed stationary phase with OV-1 on Chromosorb G/NAW 60-80 mesh size for the GC separation of aromatic hydrocarbons, heteroaromatics, alcohols, aldehydes, ketones, esters, and nitro compounds. Forty-two compounds were examined and the GC results were compared with those obtained with 3% OV-1 on Chromosorb G/NAW 60 80 mesh size under similar operating conditions. Improved resolution, peak asymmetry, theoretical plate numbers, and Kovats indices were obtained on the mixed stationary phase of 3% OV-1 + 5% PdAA2en compared to 3% OV-1 column. The stability constant (Km), enthalpies (deltaH), entropies (deltaS), and Gibbs free energies (deltaG) of the GC elution on the column (2 m x 3 mm id) packed with 3% OV-1 + 5% PdAA2en on Chromosorb G/NAW 60-80 mesh size have been calculated. Donor acceptor complexation in the gas phase is indicated by negative values of enthalpy (-deltaH) within 5.81 to 10.89 kcal/mol and Gibbs free energy (-deltaG) from 1.34 to 4.08 kcal/mol. The retention time transformations on both the phases were calculated and the ratios obtained from the stationary phases 3% OV-1 and 3% OV-1 + 5% PdAA2en were plotted on the x-axis and the y-axis, respectively, and the coefficients of determination for alkanes, aromatic and heteroaromatics, ketones, and alcohols were observed to lie within 0.5253 to 0.9742, indicative of some different kinds of interaction of the two stationary phases with the solutes. PMID- 17396593 TI - Usefulness of autoantigens depletion to detect autoantibody signatures by multiple affinity protein profiling. AB - Patients with cancer produce specific autoantibodies against protein antigens present in limited amount among a large background of immunoglobulins (Igs), nonrelevant as biomarkers, including natural antibodies. Multiple affinity protein profiling (MAPPing) that combines 2-D immunoaffinity chromatography, enzymatic digestion of the isolated proteins, and identification by MS/MS, may facilitate the identification of these so far unknown patient antibodies. The first immunoaffinity chromatography is crucial, as it is used for selectively removing proteins (autoantigens) recognized by natural antibodies. Application of this depletion step to colon cancer cell proteins is specifically described along with the identification of the natural autoantigens, as well as the coupling of this depletion step with the next steps. By enabling to separate antibody-binding proteins recognized by either natural autoantibodies or patient-specific antibodies this approach may contribute significantly towards the definition of autoantibody signatures. PMID- 17396595 TI - Effects of injection volume change on gas chromatographic sensitivity determined with two contrasting calibration approaches for volatile organic compounds. AB - In this study, the effects of injection volume change on gas chromatographic detection properties have been evaluated using gas-phase standards containing three aromatic volatile organic compounds (VOC): benzene, toluene, and xylene (commonly called BTX). To examine such effects on GC sensitivity, a series of calibration data sets were obtained using standards of three concentration values (3, 6, and 10 ppm) at each of five selected injection volumes (20 to 1000 microL). The results were initially examined in terms of the fixed standard volume (FSV) approach to allow the direct comparison of calibration patterns between different injection volumes. Identical data sets were then re-organized so that the calibration data could also be compared across variable injection volumes for a given standard concentration (at all three concentrations), i.e. by the fixed standard concentration (FSC) approach. The results of our comparative analysis between the FSV and the FSC approaches indicate that the calibration patterns of VOC are highly sensitive to changes in injection volume or injection related conditions. It is thus suggested that the former approach is more reasonable for reducing uncertainties associated with the GC-based quantification of atmospheric pollutants. PMID- 17396596 TI - Analysis of 'ARN' naphthenic acids by high temperature gas chromatography and high performance liquid chromatography. AB - Examination by high temperature GC (HTGC) of the methyl esters of the so-called 'ARN' naphthenic acids from crude oils of North Sea UK, Norwegian Sea and West African oilfields revealed the distributions of resolved 4-8 ring C80 tetra acids and trace amounts of other acids. Whilst all three oils contained apparently the same major acids, the proportions of each differed, possibly reflecting the growth temperatures of the archaebacteria from which the acids are assumed to have originated. The structures of the 4, 5, 7 and 8 ring acids are tentatively assigned by comparison with the known 6 ring acid and related natural products and an HPLC method for the isolation of the individual acids is described. ESI-MS of individual acids isolated by preparative HPLC established the elution order of the 4-8 ring acids on the HPLC and HTGC systems and revealed the presence of previously unreported acids tentatively identified as C81 and C82 7 and 8 ring analogues. PMID- 17396597 TI - Development of a quality evaluation method for Fructus schisandrae by pressurized capillary electrochromatography. AB - A pressurized CEC (pCEC) method with postcolumn detection cell had been developed for quantifying the lignans from Fructus schisandrae extracts. The effects of different experimental conditions, such as the ACN content of the mobile phase, the concentration and pH of the buffer, the applied voltage, and the supplementary pressure were studied. Five lignans (schisandrin, gomisin A, schisantherin C, deoxyschizandrin, schisandrin B) were baseline separated using a mobile phase of ACN-phosphate buffer (pH 5.4; 5 mM) (40:60 v/v) under -4 kV applied voltage. The method showed the satisfactory retention time and peak area repeatability. The calibration curves were linear in the range 50.0-1000.0 microg/mL for schisandrin, 20.0-500.0 microg/mL for gomisin A, 10.0-200.0 microg/mL for schisantherin C, 20.0-500.0 microg/mL for deoxyschizandrin, and 20.0-500.0 microg/mL for schisandrin B. The correlation coefficients were between 0.9978 and 0.9989. With this pCEC system, fingerprints of F. schisandrae were preliminarily established to distinguish two members S. chinensis (Turcz.) Baill. and S. sphenanthera Rehd. Et Wils. of F. schisandrae by characteristic peaks, and evaluate the quality of various sources of raw materials by determining the contents of the five lignans. PMID- 17396598 TI - Online preconcentration of arsenic compounds by dynamic pH junction-capillary electrophoresis. AB - An online preconcentration technique by dynamic pH junction was studied to improve the detection limit for anionic arsenic compounds by CE. The main target compound is roxarsone, or 3-nitro-4-hydroxyphenylarsonic acid, which is being used as an animal feed additive. The other inorganic and organoarsenic compounds studied are the possible biotransformation products of roxarsone. The arsenic species were separated by a dynamic pH junction in a fused-silica capillary using 15 mM phosphate buffer (pH 10.6) as the BGE and 15 mM acetic acid as the sample matrix. CE with UV detection was monitored at a wavelength of 192 nm. The influence of buffer pH and concentration on dynamic pH junction were investigated. The arsenic species focusing resulted in LOD improvement by a factor of 100-800. The combined use of C18 and anion exchange SPE and dynamic pH junction to CE analysis of chicken litter and soils helps to increase the detection sensitivity. Recoveries of spiked samples ranged between 70 and 72%. PMID- 17396599 TI - Application of micellar and microemulsion electrokinetic chromatography for characterization of gallium(III) complexes of pharmaceutical significance. AB - CE with conventional UV detection has recently been shown as a highly effective means to assaying cytotoxic gallium(III)-based compounds with regard to desirable drug-like properties such as the stability and binding to serum proteins. In this extension of that work, different CE techniques are used to further characterize a given set of gallium coordination compounds with established antiproliferating efficacy. Using free-zone CE mode, the electrophoretic profiles of complexes are recorded in order to assess their actual charge state under physiological buffer conditions. Micellar and microemulsion electrokinetic chromatographic techniques are tested as tools for the rapid estimation of the n-octanol-water partition coefficient (log P) that provides a rationale estimate of a drug's ability to cross biological membranes. A range of electrolyte buffer systems with varying (both in the nature and concentration) organic modifiers are examined to evaluate their effect on the relationship between experimental or calculated log P and the retention factors of compounds (log k'). Both methods were found to be better applicable for neutral than for cationic Ga complexes, the microemulsion mode demonstrating superior lipophilicity estimations as well as statistically meaningful log P versus log k' correlations when all the complexes were included in one regression set. PMID- 17396600 TI - Controlling the surface chemistry and chromatographic properties of methacrylate ester-based monolithic capillary columns via photografting. AB - Preparation of monolithic capillary columns for separations in the CEC mode using UV-initiated polymerization of the plain monolith followed by functionalization of its pore surface by photografting has been studied. The first step enabled the preparation of generic poly(butyl methacrylate-co-ethylene dimethacrylate) monoliths with optimized porous properties, controlled by the percentages of porogens 1-decanol and cyclohexanol in the polymerization mixture, irradiation time, and UV light intensity. Ionizable monomers [2 (methacryloyloxy)ethyl]trimethylammonium chloride or 2-acryloamido-2-methyl-1 propanesulfonic acid were then photografted onto the monolithic matrix, allowing us to control the direction of the EOF in CEC. Different strategies were applied to control the grafting density and, thereby, the magnitude of the EOF. To control the hydrophobic properties, two approaches were tested: (i) cografting of a mixture of the ionizable and hydrophobic monomers and (ii) sequential grafting of the ionizable and hydrophobic monomers. Cografting resulted in similar retention but higher EOF. With sequential grafting, more than 50% increase in retention factors was obtained and a slight decrease in EOF was observed due to shielding of the ionizable moieties. PMID- 17396601 TI - Statistical mixture design optimization of extraction media and mobile phase compositions for the characterization of green tea. AB - The influence of different solvents on the extraction medium and the RP-HPLC mobile phase composition were investigated by statistical mixture designs to optimize solvent proportions to prepare the fingerprint of a medicinal herbal extract. For modeling, the number of peaks was used as a measure of fingerprint information. The optimum compositions of solvent to extract chemical substances from green tea and for mobile phase chromatographic analysis were ethyl acetate/ ethanol/dichloromethane (20:5:75 v/v/v) and MeOH/ACN/water (7.5:57.5:35 v/v/v), respectively. This system results in 26 peaks in the chromatographic fingerprint. These results show that an incorrect choice of modifiers for mobile phase composition and solvent extraction hampers the detection of a maximum number of peaks and produces a poor chromatographic fingerprint. PMID- 17396602 TI - Determination of dopamine and serotonin in human urine samples utilizing microextraction online with liquid chromatography/electrospray tandem mass spectrometry. AB - A specific LC-MS-MS method for the determination of dopamine and serotonin (5 hydroxytryptamine; 5HT) in human urine is described. The analytes were extracted from urine and preconcentrated by microextraction in a packed syringe (MEPS). The new method is very promising, very easy to use, fully automated, of low cost, and rapid in comparison to previously used methods. The method was validated and the standard curves were evaluated by means of quadratic regression and weighted by inverse of the concentration: 1/x for the calibration range 50-4000 microg/L. The MEPS applied polymer (silica-C8) could be used more than 300 times. The extraction recovery was about 50%. The results showed close correlation coefficients (r2 > 0.999) for all analytes in the calibration range studied. The accuracy of MEPS-LC-MS-MS was 100-101% for dopamine and 99-100% for 5HT. The interday precision (n = 3 days), expressed as the RSD%, was 6.0-7.7% for dopamine and 6.1-11% for 5HT. MEPS reduced the handling time by 12 times compared to a published method. PMID- 17396608 TI - [Nursing care of patients with stomas]. PMID- 17396609 TI - [Arterial puncture for blood gases]. PMID- 17396610 TI - [Making relationship touching the heart of care]. PMID- 17396611 TI - [Transcutaneous neurostimulation: equipment and nursing practices]. PMID- 17396612 TI - [Health in the prison milieu]. PMID- 17396613 TI - [Management of medical care waste products]. PMID- 17396614 TI - [Transmitting competences developed during the care of children]. PMID- 17396615 TI - [Macrogols]. PMID- 17396616 TI - [Positron emission tomography using FDG]. PMID- 17396617 TI - Influence of systemically given placebo, trapidil and isosorbide dinitrate on norepinephrine-evoked hand vein constriction in healthy subjects. AB - Since trapidil (CAS 15421-84-8) is able to dilate human hand veins after local intravenous administration, four studies were carried out in healthy male volunteers using the dorsal hand vein compliance technique to test the influence of common systemic single doses of trapidil (200 mg orally, 100 mg intravenously) and isosorbide dinitrate (CAS 87-33-2, 20 mg orally) on norepinephrine (CAS 51-41 2)-evoked hand vein constriction in comparison with oral placebo. Oral placebo and oral trapidil were studied in a randomized double-blind cross-over design in 10 subjects aged 20 to 30 years, and oral isosorbide dinitrate and intravenous trapidil, in a randomized open cross-over design in 8 subjects aged 22 to 29 years. In the three similar studies with oral medications dose-response curves for venoconstriction by locally infused norepinephrine were established before and 1 h, 2 h and 3 h after oral medication and ED50 values of norepinephrine were calculated. The control dose-response curves and ED50 values of norepinephrine did not differ significantly. After oral placebo administration the dose-response curves of norepinephrine did not change significantly, but the ED50 of norepinephrine increased 3 h after placebo (from 12.1 to 31.7 ng/ min), indicating a lessening in norepinephrine effect at this time. After oral trapidil application the dose-response curves of norepinephrine shifted to the left compared with the pre-treatment curve (significantly 2 h after trapidil) and the corresponding curves after placebo with a significant decrease in the ED50 of norepinephrine 3 h after trapidil compared with placebo (from 31.7 to 12.6 ng/ min). After oral isosorbide dinitrate administration the dose-response curves of norepinephrine did not differ significantly from the pre-treatment curve, but they shifted to the left compared with the corresponding curves after placebo (significantly 3 h after isosorbide dinitrate). The ED50 of norephinephrine decreased significantly 2 h after isosorbide dinitrate compared with the pre treatment value (from 9.4 to 3.3 ng/min) as well as 1 h, 2 h and 3 h after isosorbide dinitrate compared with placebo (from 32.4/21.3/31.7 to 7.3/3.3/6.0 ng/min). Therefore, oral trapidil and isosorbide dinitrate did not weaken norepinephrine-evoked hand vein constriction as expected but strengthened it slightly. Intravenously given trapidil led only to an insignificant short decrease followed by an insignificant increase in permanent venoconstriction due to local norepinephrine infusion. The data suggest that after systemic administration of trapidil or isosorbide dinitrate a hand vein constriction, which could be a reflex consequence of a drug-induced decrease in central venous pressure, exceeds an only discreet direct hand vein dilation. PMID- 17396618 TI - Comparative evaluation of the leukotriene receptor antagonist pranlukast versus the steroid inhalant fluticasone in the therapy of aged patients with mild bronchial asthma. AB - A comparative study was conducted in elderly subjects with mild bronchial asthma to investigate the clinical usefulness of monotherapy with a leukotriene receptor antagonist in comparison to an inhaled corticosteroid. A total of 41 elderly patients aged 65 years or older with mild bronchial asthma, classified as being in severity step 1 and 2, were randomly assigned to the following two treatment groups: a pranlukast (CAS 103177-37-3, Onon) treatment group of 21 patients and an inhaled corticosteroid treatment group of 20 patients. Patients of the former group received pranlukast 450 mg daily and those of the latter group received fluticasone (CAS 90566-53-3) 200 microg daily for eight weeks. In the reference group, one patient was found to suffer from oral candidiasis 4 weeks after the start of the study. Therefore the evaluation was conducted on the remaining 19 participants. The evaluation parameters examined were obtained by keeping an asthma diary, determinations of PEF (peek expiratory flow), use frequency of beta2 stimulants, changes in symptom scores, and medication compliance. Further, measured before and after therapy were the ratio of peripheral blood eosinophils counts, serum ECP (eosinophils cationic protein), ECP levels induced sputum, and forced expiratory volume in one second (FEV1.0). As a result, in the time-course changes of symptoms scores and morning PEF, swift improvement was noted in the pranlukast group. Further, in the variables such as use frequency of beta2 stimulants, serum ECP levels, ECP levels induced sputum, and FEV1.0, an almost comparable level of improvement to the fluticasone group was demonstrated. From the above results, it was deemed that in elderly patients with mild bronchial asthma classified as steps 1 and 2, the pranlukast monotherapy, with superior medication compliance to inhaled therapy, would produce an equivalent level of clinical efficacy to the monotherapy with inhaled corticosteroid (fluticasone 200 microg daily). PMID- 17396619 TI - Effects of imidafenacin (KRP-197/ONO-8025), a new anti-cholinergic agent, on muscarinic acetylcholine receptors. High affinities for M3 and M1 receptor subtypes and selectivity for urinary bladder over salivary gland. AB - Imidafenacin (CAS 170105-16-5, KRP-197, ONO-8025) is an antagonist for the muscarinic acetylcholine (ACh) receptor currently under development for the treatment of overactive bladder. Affinities of imidafenacin and other drugs for muscarinic ACh receptor subtypes were investigated by examining inhibitory effects on ACh release in the rat urinary bladder and K+ efflux in the rat salivary gland in functional and binding assays. In the functional assay, imidafenacin had higher affnities for M3 and M1 receptors than for the M2 receptor. In contrast, metabolites of imidafenacin (M-2, M-4 and M-9) had low affinities for muscarinic ACh receptor subtypes. Darifenacin had selectivity for the M3 receptor, while propiverine, tolterodine and oxybutynin had no selectivity for muscarinic ACh receptors. In carbamylcholine (CCh)-induced contraction in the urinary bladder, imidafenacin, propiverine, tolterodine and oxybutynin had affinities similar to those for the M3 receptor in the ileum. In the binding assay for human muscarinic ACh receptor subtypes, imidafenacin had higher affinities for m3 and m1 receptors than for m2 receptor, but tolterodine had no selectivity for m1, m2 and m3 receptors. In ACh release in the urinary bladder, inhibitory effects of imidafenacin, tolterodine, oxybutynin and darifenacin seemed to be partially mediated by the M1 receptor. In ACh-induced and electrical stimulation-induced K+ efflux from the salivary gland, inhibitory effects (IC50) of imidafenacin, propiverine, tolterodine, oxybutynin and darifenacin might be closely related to those for the M3 receptor in the ileum. These results suggest that imidafenacin more strongly antagonizes cholinomimetics on M3 and M1 receptors than on the M2 receptor. Moreover, imidafenacin seems to inhibit the contraction of the bladder smooth muscle by mediating antagonism to the M3 receptor and to regulate ACh release by mediating prejunctional facilitatory M1 receptor. Imidafenacin also inhibited K+ efflux from the salivary gland mainly by mediating the M3 receptor. Therefore, imidafenacin will have higher affinities for M3 and M1 receptors and higher selectivity for the urinary bladder than for the salivary gland. PMID- 17396620 TI - Bioequivalence assessment of two capsule formulations of omeprazole in healthy volunteers. AB - A randomized, single-dose, crossover study was conducted to assess the bioavailability of two omeprazole (CAS 73590-58-6) capsule formulations, Emilok (test) and a commercially available original preparation (reference), under fasting conditions. A 20 mg dose of each formulation was administered to 36 healthy male volunteers with one-week washout period, 17 blood samples were collected over 12 h, plasma omeprazole concentrations were determined by a locally validated high performance liquid chromatography (HPLC) assay, and omeprazole pharmacokinetic parameters were analyzed by the standard non compartmental method. Mean +/- SD of Cmax, Tmax, AUC(0 --> t), AUC(0 --> infinity), and t1/2 were 0.41 +/- 0.21 and 0.48 +/- 0.27 microg/ml, 1.98 +/- 1.02 and 1.63 +/- 0.78 h, 0.95 +/- 0.78 and 1.00 +/- 0.90 microg x h/ml, 0.99 +/- 0.81 and 1.04 +/- 0.95 microg x h/ml, and 1.30 +/- 0.64 and 1.14 +/- 0.61 h for the test and reference formulations, respectively. The parametric 90% confidence intervals on mean difference between log-transformed values of the two formulations were within the acceptable bioequivalence range of 80% to 125% for AUC(0 --> t) and AUC(0 --> infinity) (88.63% to 104.98%, and 91.71% to 106.86%, respectively) but not for Cmax (76.27% to 103.63%). ANOVA revealed significant subject's effect for AUC(0 --> t), AUC(0 --> infinity), Cmax, and t1/2 with a ratio of the inter-subject to intra-subject coefficient of variation of 3.66, 3.92, 1.25, and 1.46, respectively. The results confirm the presence of marked individual variations in the pharmacokinetics of omeprazole and indicate that the two formulations are equivalent in relation to the extent but not the rate of absorption. PMID- 17396621 TI - Pharmacokinetics of tilidine and naloxone in patients with severe hepatic impairment. AB - The objective of the present study was to evaluate the pharmacokinetics of tilidine (CAS 20380-58-9), naloxone (CAS 465-65-6) and tilidine metabolites after administration of a single oral dose of a solution containing 100 mg tilidine hydrochloride and 8 mg naloxone hydrochloride (equivalent to 1.44 ml Valoron N solution) to patients with severe hepatic impairment. The investigation was carried out as an open single-dose study in 8 patients suffering from liver cirrhosis. Patients qualified for study enrollment if they had a Child-Pugh score of > or = 7 and a mono-ethyl-glycine-xylidide (MEGX) 15-min test value < 50 ng/ ml. Blood samples were taken over a period of 28 h and analyzed for the prodrug tilidine, its active metabolite nortilidine, bisnortilidine, and naloxone (total and non-glucuronidated fraction). Pharmacokinetic parameters were compared with data from a previous study performed in healthy volunteers. Tilidine, nortilidine and unconjugated naloxone pharmacokinetic parameters showed a high variability between patients. Compared to previous results obtained in healthy volunteers, maximum plasma concentration (Cmax) of nortilidine was reduced by 44%, whereas elimination half-life (t1/2) was prolonged by factor 2. The area under the curve (AUC) showed a slight reduction of approximately 20%. For total naloxone, no relevant change was observed. However, in contrast to the results obtained in healthy subjects, unconjugated naloxone could be measured in plasma from patients with cirrhosis, possibly due to a reduced glucuronidation capacity of the liver in these patients. In conclusion, severe hepatic impairment has a relatively minor influence on the exposure (AUC) to the active metabolite of tilidine (i.e., nortilidine). However, a straightforward interpretation of the results was confounded by pronounced variability in nortilidine pharmacokinetics. In individual patients with severely affected liver function, satisfactory analgesia with tilidine/naloxone oral solution might not be achieved because of insufficient formation of nortilidine and insufficient inactivation of naloxone. PMID- 17396622 TI - Anti-retroviral activity of GMP-grade stampidine against genotypically and phenotypically nucleoside reverse transcriptase inhibitor resistant recombinant human immunodeficiency virus. An in vitro study. AB - The in vitro potency of GMP-grade stampidine (CAS 217178-62-6) was examined against 3 clinical HIV-1 isolates and 6 recombinant HIV-1 clones with multi-NRTI 'resistance (NRTI: nucleoside reverse transcriptase inhibitors). GMP-grade stampidine active drug substance (Lot #'s MPR-M0008.00-01 and MPR-M0008.01-01) as well as GMP-grade stampidine extracted from the clinical stampidine capsules (GMP Grade Clinical Batch, Pharmaceutical Service Lot Number 159I0601) were highly potent and exhibited nanomolar IC50 values against clinical HIV-1 isolates as well as recombinant HIV-1 clones with multi-NRTI resistance containing common patterns of reverse transcriptase mutations responsible for NRTI resistance. PMID- 17396623 TI - Tandem mass spectrometry (LC-MS-MS): a predominant role in bioassays for pharmacokinetic studies. AB - Plasma concentrations of drugs and drug metabolites in bioavailability studies are routinely bioassayed with high performance liquid chromatography (HPLC), gas chromatography (GC), and liquid chromatography coupled with tandem mass spectrometry (LC-MS-MS) methods. In the 1980s and 1990s, HPLC had achieved a relevant role compared to GC and other techniques in pharmacokinetic bioassays. However, in the last few years the LC-MS-MS technique, known as tandem mass spectrometry, has attained a predominant role over all other techniques. This is because it requires a less amount of matrix, possesses better specificity and sensitivity, and requires a shorter operating time. In the experience of the authors, LC-MS-MS in fact requires on average 3-4 fold shorter time to complete a bioequivalence study when performed by one operator than does HPLC. The higher cost of the apparatus and technical assistance of LC-MS-MS are fully compensated by the shorter operating time. Part or most of the HPLC apparatuses have been or are being replaced by LC-MS-MS systems in laboratories operating in clinical and pre-clinical pharmacokinetics and, to a large extent, in those involved in assessing permeability in screening procedures of new chemical entities. This paper analyses the growing development of the LC-MS-MS technique and compares four couples of methods validated with HPLC or GC versus LC-MS-MS, giving analytical details of the two approaches. PMID- 17396624 TI - Optimist by a landslide. PMID- 17396625 TI - The IPCC fourth assessment. PMID- 17396626 TI - Arsenic in Katrina's wood debris. PMID- 17396627 TI - The risk of PBDEs in dust. PMID- 17396628 TI - Dioxins and PCBs in rural areas. PMID- 17396629 TI - Antimony levels in bottled water. PMID- 17396630 TI - Climate change could offset ozone cleanup. PMID- 17396631 TI - Debate over lead in air. PMID- 17396632 TI - Unreliable water-quality forecasts at mines. PMID- 17396633 TI - CO2 in the Supreme Court. PMID- 17396634 TI - Lubes from vegetable oils become affordable. PMID- 17396635 TI - Known and unknown organic constituents in the Earth' s atmosphere. PMID- 17396636 TI - Economic input-output life-cycle assessment of trade between Canada and the United States. AB - With increasing trade liberalization, attempts at accounting for environmental impacts and energy use across the manufacturing supply chain are complicated by the predominance of internationally supplied resources and products. This is particularly true for Canada and the United States, the world's largest trading partners. We use an economic input-output life-cycle assessment (EIO-LCA) technique to estimate the economy-wide energy intensity and greenhouse gas (GHG) emissions intensity for 45 manufacturing and resource sectors in Canada and the United States. Overall, we find that U.S. manufacturing and resource industries are about 1.15 times as energy-intensive and 1.3 times as GHG-intensive as Canadian industries, with significant sector-specific discrepancies in energy and GHG intensity. This trend is mainly due to a greater direct reliance on fossil fuels for many U.S. industries, in addition to a highly fossil-fuel based electricity mix in the U.S. To account for these differences, we develop a 76 sector binational EIO-LCA model that implicitly considers trade in goods between Canada and the U.S. Our findings show that accounting for trade can significantly alter the results of life-cycle assessment studies, particularly for many Canadian manufacturing sectors, and the production/consumption of goods in one country often exerts significant energy- and GHG-influences on the other. PMID- 17396637 TI - Quantities of arsenic-treated wood in demolition debris generated by Hurricane Katrina. AB - The disaster debris from Hurricane Katrina is one of the largest in terms of volume and economic loss in American history. One of the major components of the demolition debris is wood waste of which a significant proportion is treated with preservatives, including preservatives containing arsenic. As a result of the large scale destruction of treated wood structures such as electrical poles, fences, decks, and homes a considerable amount of treated wood and consequently arsenic will be disposed as disaster debris. In this study an effort was made to estimate the quantity of arsenic disposed through demolition debris generated in the Louisiana and Mississippi area through Hurricane Katrina. Of the 72 million cubic meters of disaster debris generated, roughly 12 million cubic meters were in the form of construction and demolition wood resulting in an estimated 1740 metric tons of arsenic disposed. Management of disaster debris should consider the relatively large quantities of arsenic associated with pressure-treated wood. PMID- 17396638 TI - A general indication of the Contemporary background levels of PCDDs, PCDFs, and coplanar PCBs in the ambient air over rural and remote areas of the United States. AB - Long-term measurements of the atmospheric concentrations of PCDDs, PCDFs, and coplanar PCBs were taken in rural and remote areas of the United States by the National Dioxin Air Monitoring Network (NDAMN). A total of 21 quarterly sampling moments occurred from June, 1998 to December, 2002 at 34 locations geographically distributed throughout the United States. Sampling sites were located in rural and remote areas to obtain background air concentrations of dioxin-like compounds. Results were reported as the toxic equivalent (TEQ) of the mix of PCDDs/ PCDFs (TEQDF) and the mix of coplanar PCBs (TEQPCB). At the studied rural sites the mean annual TEQDF for each of the NDAMN sampling years was 10.43, 11.39, 10.40, and 10.47 fg m(-3) for 1999, 2000, 2001, and 2002, respectively. There was no statistically significant difference in the rural mean TEQDF air concentrations across the sampling years (at 0.05 level of significance), although the mean concentration in sampling year 2000 increased 10% relative to the other sampling years. The 95th percent confidence interval of TEQDF air concentrations in rural areas of the United States is from 6.4 to 15.4 fg m(-3), indicating there is a 95% probability that the true mean falls within this range. Mean annual atmospheric concentrations (TEQDF) at the remote sites were 1.41, 0.99, 0.7, and 1.07 fg m(-3) in 1999, 2000, 2001, and 2002, respectively, The 95th percent confidence interval of TEQDF air concentrations suggest that the true mean annual atmospheric TEQDF concentration in remote areas of the United States is between 0.1 and 3 fg m(-3). The remote sites have average air TEQDF concentrations that are approximately 10 times lower than those of the rural sites. For the rural sites, there was close agreement in the mean annual air concentrations of coplanar PCBs (TEQPCB) among the years 1999, 2000, 2001, and 2002 (i.e., 0.62, 0.69, 0.59, and 0.7 fg m(-3), respectively). However, as is the case with PCDDs/PCDFs, there was a marked increase (i.e., approximately equal to 13%) in the annual rural mean air concentration in 2000 as compared to the other sampling years. The confidence intervals across the NDAMN sampling years suggests a 95% probability that mean TEQPcB atmospheric concentrations in rural and remote areas of the United States are within the range of 0.5-0.9 fg m(-3) and 0.1-0.5 fg m(-3), respectively. The congener distributional patterns of PCDDs/PCDFs in air were relatively constant at all locations, and match the profile of urban air. We propose the hypothesis that urban areas are regional sources of PCDDs/PCDFs and are affecting atmospheric levels in rural and remote areas of the United States. PMID- 17396639 TI - Sesquiterpene emissions from pine trees--identifications, emission rates and flux estimates for the contiguous United States. AB - Biogenic volatile organic compound (BVOC) emissions were studied using vegetation enclosure experiments. Particular emphasis was given to sesquiterpene compounds (SQT), although monoterpenes (MT) were also characterized. SQT were detected in emissions from seven (out of eight) pine species that were examined. Thirteen SQT compounds were identified; the most abundant ones were beta-caryophyllene, alpha bergamotene, beta-farnesene, and alpha-farnesene, with emission rates increasing exponentially with temperature. Regression analysis yielded exponential dependencies of both MT and SQT emissions on temperature of the form E = E0 x exp(beta(T - T0)). This resulted in SQT basal emission rates (E0 defined at T0 = 30 degrees C) ranging between <4 and 620 ng (carbon) gdw(-1) h(-1) (gdw = gram dry weight). The average value of the exponential temperature response factor beta for SQT emissions, taken from all experiments, was 0.17 degree C(-1), whereas the value for monoterpenes was 0.11 degrees C(-1). The average, total SQT emissions from pines were estimated to be 9, 16, and 29% of the MT emissions at 20, 30, and 40 degrees C respectively. The emission factors and beta-factors determined from these measurements were used to estimate pine tree MT and SQT emission distributions for the contiguous United States using MEGAN (model of emissions of gases and aerosols from nature, Guenther et al., 2006). SQT fluxes reaching 10-40 mg m(-2) for the month of July were estimated for extensive areas of most western and southern U.S. states. PMID- 17396640 TI - Spatial distribution of perfluoroalkyl contaminants in lake trout from the Great Lakes. AB - Individual whole body homogenates of 4 year old lake trout (Salvelinus namaycush) samples collected in 2001 from each of the Great Lakes were extracted using a novel fluorophilicity cleanup step and analyzed for perfluoroalkyl compounds (PFCs). Standard addition and internal standardization were used for quantification. Results were reported (+/- SE) for perfluorinated carboxylates (PFCAs), perfluorinated sulfonates (PFSAs), and unsaturated fluorotelomer carboxylates (8:2 and 10:2 FTUCA). The lowest average concentration of sigmaPFC was found in samples from Lake Superior (13+/-1 ng g(-1)), while the highest average concentration was found in samples from Lake Erie (152+/-14 ng g(-1)). Samples from Lake Ontario (60+/-5 ng g(-1)) and Lake Huron (58 +/-10 ng g(-1)) showed similar average sigmaPFC concentrations, although the perfluorinated sulfonate/carboxylate ratios were different. The major perfluoroalkyl contaminant observed was perfluorooctane sulfonate (PFOS) with the highest concentration found in samples from Lake Erie (121+/-14 ng g(-1)), followed by samples from Lake Ontario (46+/-5 ng g(-1)), Lake Huron (39 +/-10 ng g(-1)), Lake Michigan (16+/-3 ng g(-1)), and Lake Superior (5+/-1 ng g(-1)). Perfluorodecane sulfonate (PFDS) was detected in 89% of the samples, with the highest concentration in Lake Erie samples (9.8+/-1.6 ng g(-1)), and lowest concentration in samples from Lake Superior (0.7 +/- 0.1 ng g(-1)). Statistically significant correlations were observed between PFOS and PFDS concentrations, and PFOS concentration and body weight, respectively. The PFCAs were detected in all samples, with the highest total average concentration in samples from Lake Erie (19 ng g(-1)), followed by samples from Lake Huron (16 ng g(-1)), Lake Ontario (10 ng g(-1)), Lake Michigan (9 ng g(-1)) and Lake Superior (7 ng g(-1)). The compounds with significant contributions to the sigmaPFCA concentrations were PFOA and C9-C13-PFCAs. The 8:2 FTUCA was detected at concentrations ranging between 0.1 and 0.2 ng g-1, with the highest level in samples showing also elevated concentrations of PFOA (4.4 ng g( 1) for Lake Michigan vs 1.5 ng g(-1) for all other samples). The 10:2 FTUCA was detected only in 9% of all samples (nd, 45 pg g(-1)). For those PFCs where we determined lake water concentrations, the highest log BAFs were calculated for PFOS (4.1), PFDA (3.9), and PFOSA (3.8). PMID- 17396641 TI - Contamination of bottled waters with antimony leaching from polyethylene terephthalate (PET) increases upon storage. AB - Antimony concentrations were determined in 132 brands of bottled water from 28 countries. Two of the brands were at or above the maximum allowable Sb concentration for drinking water in Japan (2 microg/L). Elevated concentrations of Sb in bottled waters are due mainly to the Sb2O3 used as the catalyst in the manufacture of polyethylene terephthalate (PET(E)). The leaching of Sb from PET(E) bottles shows variable reactivity. In 14 brands of bottled water from Canada, Sb concentrations increased on average 19% during 6 months storage at room temperature, but 48 brands of water from 11 European countries increased on average 90% under identical conditions. A mineral water from France in PET(E), purchased in Germany, yielded 725 ng/L when first tested, but 1510 ng/L when it was stored for 6 months at room temperature; the same brand of water, purchased in Hong Kong, yielded 1990 ng/L Sb. Pristine groundwater containing 1.7+/-0.4 ng/L Sb (n = 6) yielded 26.6+/-2.3 ng/L Sb (n = 3) after storage in PET(E) bottles from Canada for 6 months versus 281+/-38 ng/L Sb (n = 3) in PET(E) bottles from Germany. Tap water bottled commercially in PET(E) in December 2005 contained 450+/-56 ng/L Sb (n = 3) versus 70.3+/-0.3 ng/L Sb (n = 3) when sampled from a household faucet in the same village (Bammental, Germany), and 25.7+/-1.5 ng/L Sb (n = 3) from a local artesian flow. PMID- 17396642 TI - Biogeochemistry of organic and inorganic arsenic species in a forested catchment in Germany. AB - Little is known about the fate and behavior of diffuse inputs of arsenic (As) species in forested catchments which often are the sources of drinking water. The objective of this study was to investigate the mobility and transformation of different As species in forest ecosystems to assess the environmental risk related to the diffuse pollution of As. We determined concentrations and fluxes in precipitation, litterfall, soil solutions (Oa horizon and 20- and 90-cm depth), and runoff of organic and inorganic As species and Astotal in a forest ecosystem in NE-Bavaria, Germany. The concentrations of Astotal were mostly <1 microg As L(-1) in aqueous samples and were highest in forestfloor percolates (7.6 microg As L(-1)). In litterfall, the concentrations of As species never exceeded 0.1 microg As g(-1). Arsenate and arsenite were the prevalent As species in all samples. Organic As species, comprising monomethylarsonic acid, dimethylarsinic acid, trimethylarsine oxide, arsenobetaine, and three unidentified organic As species, were mostly found in throughfall reaching up to 45% of Astotal. The total deposition of Astotal (calculated as throughfall + litterfall) was 5.6 g As ha(-1) yr(-1) with 16% contribution of litterfall. The annual Astotal fluxes were 30 g As ha(-1) yr(-1) for forest floor percolates, 8.0 g As ha(-1) yr(-1) at 20-cm soil depth, and 1.4 g As ha(-1) yr(-1) at 90-cm soil depth. The annual runoff of Astotal from the catchment amounted to 3.8 g As ha( 1) yr(-1). The annual fluxes of total organic As species was highest in total deposition (1.1 g As ha(-1) yr(-1)) and decreased largely with depth in the soil profile. The annual runoff of total organic As species was only 0.08 g As ha(-1) yr(-1). Significant correlations in soil solutions and runoff were found between Astotal and dissolved organic C and Fe. Correlations between Astotal concentrations in runoff and water fluxes were seasonally dependent and with a steeper slope in the growing season than in the dormant season. The elevated concentrations of organic As species in throughfall indicate microbial methylation of As in the phyllosphere, but no evidence for methylation in the soil was found. The mass balance of the catchment points out the strong retention and probable degradation or oxidation of organic As species and arsenite but also to mobilizable pools of Astotal and arsenate. The forest floor is presently a source, whereas the mineral soil is a sink for Astotal and arsenate. The As concentrations in runoff seem to be controlled by As mobilization from forest floor and riparian wetland soils during heavy rain events and superficial flow. The risk for excessment of the drinking water threshold concentrations of As in runoff and soil solutions is considered low at our site. PMID- 17396643 TI - Effects of channel restoration on water velocity, transient storage, and nutrient uptake in a channelized stream. AB - Channel design is an important component of stream restoration, but little is known of the interplay between hydrogeomorphic features and ecosystem processes within designed channels. Water velocity, transient storage, and nutrient uptake were measured in channelized (prerestoration) and naturalized (postrestoration) reaches of a 1-km segment of Wilson Creek (KY) to assess the effects of restoration on mechanisms of nutrient retention. Stream restoration decreased flow velocity and reduced the downstream transport of nutrients. Median travel time was 50% greater in the restored channel due to lower reach-scale water velocity and the longer length of the meandering channel. Transient storage and the influence of transient storage on travel time were largely unaffected except in segments where backwater areas were created. First-order uptake rate coefficients for N and P were 30- and 3-fold higher (respectively) within the restored channel relative to its channelized state. Changes in uptake velocities were comparatively small, suggesting that restoration had little effect on biochemical demand. Results from this study suggest that channel naturalization enhances nutrient uptake by slowing water velocity. Solute injection experiments revealed differences in the functional properties of channelized, restored, and reference streams and provided a means for quantifying benefits associated with restoration of ecosystem services. PMID- 17396644 TI - Diagnostic model evaluation for carbonaceous PM2.5 using organic markers measured in the southeastern U.S. AB - Summertime concentrations of fine particulate carbon in the southeastern United States are consistently underestimated by air quality models. In an effort to understand the cause of this error, the Community Multiscale Air Quality model is instrumented to track primary organic and elemental carbon contributions from fifteen different source categories. The model results are speciated using published source profiles and compared with ambient measurements of 100 organic markers collected at eight sites in the Southeast during the 1999 summer. Results indicate that modeled contributions from vehicle exhaust and biomass combustion, the two largest sources of carbon in the emission inventory, are unbiased across the region. In Atlanta, good model performance for total carbon (TC) is attributed to compensating errors: overestimation of vehicle emissions with underestimations of other sources. In Birmingham, 35% of the TC underestimation can be explained by deficiencies in primary sources. Cigarette smoke and vegetative detritus are not in the inventory, but contribute less than 3% of the TC at each site. After the model results are adjusted for source-specific errors using the organic-marker measurements, an average of 1.6 microgC m(-3) remain unexplained. This corresponds to 26-38% of ambient TC concentrations at urban sites and up to 56% at rural sites. The most likely sources of unexplained carbon are discussed. PMID- 17396645 TI - Human exposure to PBDEs: associations of PBDE body burdens with food consumption and house dust concentrations. AB - This study was designed to determine the body burden of polybrominated diphenyl ethers (PBDEs) among first-time mothers in the Greater Boston, Massachusetts area and to explore key routes of exposure. We collected breast milk samples from 46 first-time mothers, 2-8 weeks after birth. We also sampled house dust from the homes of a subset of participants by vacuuming commonly used areas. Data on personal characteristics, diet, home furniture, and electrical devices were gathered from each participant using a questionnaire. Breast milk and dust samples were analyzed for PBDEs using gas chromatography/ mass spectrometry. PBDE concentrations were log-normally distributed in breast milk and dust. We found statistically significant, positive associations between PBDE concentrations in breast milk and house dust (r = 0.76, p = 0.003, not including BDE-209), as well as with reported dietary habits, particularly the consumption of dairy products (r = 0.41, p = 0.005) and meat (r = 0.37, p = 0.01). Due to low detection rates, it was not possible to draw conclusions about the association between BDE-209 in milk and dust. Our results support the hypothesis that the indoor environment and diet both play prominent roles in adult human exposure to PBDEs. PMID- 17396646 TI - Synthesis and characterization of 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE) and structurally related compounds evidenced in seal blubber and brain. AB - The unknown compound UBC-1 previously described as the major organobromine contamination in the blubber extract of a hooded seal (Cystophora cristata) from the Barents Sea was identified as 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE). DPTE, which is the main component of the brominated flame retardant (BFR) Bromkal 73-5 PE, was synthesized by electrophilic addition of bromine to allyl 2,4,6-tribromophenyl ether (ATE). The chirality of DPTE was proven by gas chromatographic enantioseparation of the synthesized racemate. On the basis of GC/ECNI-MS ion chromatograms (m/z79 and 81), DPTE was the dominating organobromine compound in blubber and brain samples of hooded seals and harp seals (Phoca groenlandica) from the Barents and Greenland Seas. The concentrations of DPTE in blubber and brain were up to 470 and 340 microg/kg wet weight. Next to DPTE, the natural dibromo-trichloromonoterpene (MHC-1), the anthropogenic BDE 47 and BDE 99, as well as ATE, 3,5-dibromo-2-(2',4'-dibromo) phenoxyanisole (6-MeO-BDE 47), 2-bromoallyl-2,4,6-tribromophenyl ether (BATE), and 4,6-dibromo-2-(2',4'-dibromo)-phenoxyanisole (2'-MeO-BDE 68) were present with decreasing relevance. BATE, which was detected for the first time in environmental samples, was synthesized from DPTE by E2 elimination. In brain samples of the harp seals, DPTE, ATE, and BATE were the most abundant organobromine compounds, whereas polybrominated diphenyl ethers (PBDEs) and MHC-1 were virtually absent. This indicated that DPTE, ATE, and BATE were able to penetrate the blood-brain barrier. The general co-occurrence of ATE and BATE in samples contaminated with DPTE support the hypothesis that these compounds are biotransformation products of DPTE. Anaerobic transformation studies of DPTE with super-reduced corrinoids resulted in the formation of ATE. Furthermore, 2,4,6 tribromophenol (TBP) and two other unknown minor transformation products were detected. PMID- 17396647 TI - Ion-specific isotopic fractionation of molybdenum during diffusion in aqueous solutions. AB - Experiments modeling diffusion of Mo in aqueous solutions have been performed and, using multicollector ICP-MS, the ratios of the diffusivities of Mo isotopes, D97Mo/ D95Mo, in aqueous solutions have been determined. Diffusion of MoO42- ions in solution was concomitant with Mo isotopic fractionation with D97Md/D95Mo = 0.99988+/-0.00004 (2sigma for n = 3). In contrast, during diffusion of Mo polyanions, such as Mo70246- and Mo8O264-, no measurable isotope fractionation has been found with D97Mo/D95MO = 1.00000 +/- 0.00002 (2sigma for n = 3). These results indicate the need for due consideration to Mo speciation when attempting to interpret the role of diffusive fluxes in the formation of Mo isotopic signatures in nature. They also raise the possibility that the various chemical forms of other transition metals may be characterized by species-specific isotopic fractionation effects during physicochemical reactions. PMID- 17396648 TI - Production of hydrated electrons from photoionization of dissolved organic matter in natural waters. AB - Under UV irradiation, an important primary photochemical reaction of colored dissolved organic matter (CDOM) is electron ejection to produce hydrated electrons (e-aq). The efficiency of this process has been studied in both fresh water and seawater samples with both steady-state scavenger (S-SS) and time resolved laser flash photolysis (LFP) methods. However, the apparent quantum yields (AQYs) of e-aq for the same samples using the two methods differ by as much as a factor of 100, necessitating a closer re-examination of how the process is measured. We developed a highly sensitive multipass LFP apparatus that allows detection of transient species at very low and variable UV irradiation intensities. Under single-photon conditions, we measured the AQY of e-aq from Laurentian fulvic acid as 1.3 x 10(-4), and set the upper limit for other CDOM samples at 6 x 10(-5), bringing the LFP results into agreement with those from S SS methods. We also examined the ionization at elevated irradiation intensities and clearly demonstrated that multiphoton ionization occurs at intensities well below those usually used in LFP experiments, but well above those likely to occur at the earth's surface. This multiphoton ionization is probably the cause of the high AQYs reported by earlier LFP work. In addition, we also observed in real time other photochemical reactions, such as triplet quenching and bleaching, in the single photon regime. PMID- 17396649 TI - Processes at the sediment water interface after addition of organic matter and lime to an Acid Mine Pit Lake mesocosm. AB - A strategy to neutralize acidic pit lakes was tested in a field mesocosm of 4500 m(3) volume in the Acidic Pit Mine Lake 111 in Germany. Carbokalk, a byproduct from sugar production, and wheat straw was applied near to the sediment surface to stimulate in lake microbial alkalinity generation by sulfate and iron reduction. The biogeochemical processes at the sediment-water interface were studied over 3 years by geochemical monitoring and an in situ microprofiler. Substrate addition generated a reactive zone at the sediment surface where sulfate and iron reduction proceeded. Gross sulfate reduction reached values up to 10 mmol m(-2) d(-1). The neutralization rates between 27 and 0 meq m(-2) d(-1) were considerably lower than in previous laboratory experiments. The precipitation of ferric iron minerals resulted in a growing acidic sediment layer on top of the neutral sediment. In this layer sulfate reduction was observed but iron sulfides could not precipitate. In the anoxic sediment H2S was oxidized by ferric iron minerals. H2S partly diffused to the water column where it was oxidized. As a result the net formation of iron sulfides decreased after 1 year although gross sulfate reduction rates continued to be high. The rate of iron reduction exceeded the sulfate reduction rate, which resulted in high fluxes of ferrous iron out of the sediment. PMID- 17396650 TI - Degradation of disinfection byproducts by carbonate green rust. AB - Disinfection byproducts (DBPs) in drinking water flowing through corroded iron or steel pipes may encounter carbonate green rust (GR(CO32-)), a mixed Fe(II)/Fe(lll) hydroxide mineral and potent reductant. This research was performed to investigate the kinetics and pathways of the degradation of selected halogenated DBPs in the presence of GR(C032-). Trichloronitromethane was rapidly degraded to methylamine via sequential hydrogenolysis followed by nitro reduction. Haloacetic acids reacted solely via sequential hydrogenolysis. Trichloroacetonitrile, 1,1,1-trichloropropanone, and trichloroacetaldehyde hydrate were transformed via hydrolysis and hydrogenolysis. Chloroform was unreactive over 300 h. The buffer identity affected reductive dehalogenation rates of DBPs, with faster rates in MOPS buffer than in carbonate buffer, the latter being representative of the buffer in drinking water systems. GR(CO32-) was unstable in both buffers and transformed to magnetite within 48 h. Thus, slower reacting compounds (half life >3 hours) were transformed by a combination of minerals. Reductive dehalogenation kinetics were influenced by DBP chemical structure and correlated with one-electron reduction potential. PMID- 17396651 TI - Fluid and porous media property effects on dense nonaqueous phase liquid migration and contaminant mass flux. AB - The effects of fluid and porous media properties on dense nonaqueous phase liquid (DNAPL) migration and associated contaminant mass flux generation were evaluated. Relationships between DNAPL mass and solute mass flux were generated by measuring steady-state mass flux following stepwise injection of perchloroethylene (PCE) into flow chambers packed with homogeneous porous media. The effects of fluid properties including density and interfacial tension (IFT), and media properties including grain size and wettability were evaluated by varying the density contrast and interfacial tension properties between PCE and water, and by varying the porous media mean grain diameter and wettability characteristics. Contaminant mass flux was found to increase as grain size decreased, suggesting enhanced lateral and vertical DNAPL spreading with higher fluid entry pressure. Mass flux showed a slight increase as the DNAPL approached neutral buoyancy, likely due to enhanced vertical spreading above the injection point. DNAPL spatial distribution and contaminant mass flux were only minimally affected by IFT and by intermediate level wettability changes, but were dramatically affected by wettability reversal. The relationship between DNAPL loading and flux generation became more linear as grain size decreased and density contrast between fluids decreased. These results imply that capillary flow characteristics of the porous media and fluid properties will control mass flux generation from source zones. PMID- 17396652 TI - Hydroxydicarboxylic acids: markers for secondary organic aerosol from the photooxidation of alpha-pinene. AB - Detailed organic analysis of fine (PM2.5) rural aerosol collected during summer at K-puszta, Hungary from a mixed deciduous/coniferous forest site shows the presence of polar oxygenated compounds that are also formed in laboratory irradiated alpha-pinene/NOx/air mixtures. In the present work, two major photooxidation products of alpha-pinene were characterized as the hydroxydicarboxylic acids, 3-hydroxyglutaric acid, and 2-hydroxy-4 isopropyladipic acid, based on chemical, chromatographic, and mass spectral data. Different types of volatile derivatives, including trimethylsilyl ester/ether, methyl ester trimethylsilyl ether, and ethyl ester trimethylsilyl ether derivatives were measured by gas chromatography/mass spectrometry (GC/MS), and their electron ionization (El) spectra were interpreted in detail. The proposed structures of the hydroxydicarboxylic acids were confirmed or supported with reference compounds. 2-Hydroxy-4-isopropyladipic acid formally corresponds to a further reaction product of pinic acid involving addition of a molecule of water and opening of the dimethylcyclobutane ring; this proposal is supported by a laboratory irradiation experiment with alpha-pinene/NOJ0 air. In addition, we report the presence of a structurally related minor alpha-pinene photooxidation product, which was tentatively identified as the C7 homolog of 3-hydroxyglutaric acid, 3-hydroxy-4,4-dimethylglutaric acid. The detection of 2-hydroxy-4 isopropyladipic acid in ambient aerosol provides an explanation for the relatively low atmospheric concentrations of pinic acid found during daytime in forest environments. PMID- 17396653 TI - Lack of enantioselective microbial degradation of chlordane in Long Island Sound sediment. AB - Numerous studies have examined the enantiomeric compositions of trans- and cis chlordane in soils (agricultural, background, and house foundation soils) and in the atmosphere. In contrast, little is known aboutthe enantiomeric compositions of chlordane in sediment. In this work, surficial sediments and sediment cores were collected at various sites in Long Island Sound (LIS) previously surveyed by the National Oceanic and Atmospheric Administration's (NOAA) National Status and Trends (NS&T) Program. Archived surficial sediments at selected sites were acquired from the NS&T Specimen Bank. The chlordanes were racemic or nearly racemic in most archived and recently collected sediments, indicating thatthe enantiomeric compositions of the sources of chlordane to LIS sediment did not change in the past two decades, and that house foundation soils are likely the major source of chlordanes to LIS. Invariant enantiomeric compositions temporally in surficial sediments and at different depths in sediment cores clearly indicate the lack of enantioselective biodegradation in LIS sediment, in striking contrast to the widely observed enantioselective biodegradation of chlordanes in soils. PMID- 17396654 TI - Geochemical modulation of bioavailability and toxicity of nitroaromatic compounds to aquatic plants. AB - Nitroaromatic compounds (NACs) are prominent soil and sediment contaminants that are strongly adsorbed by smectites at extents that depend on hydration properties of the exchangeable cation. Potassium smectites adsorb nitroaromatics much more strongly than calcium smectites, so that adjustment of K+ versus Ca2+ occupation on cation exchange sites in smectites can be used to modulate the retention and release of nitroaromatics. We suggest that this modulation can be used to advantageously manage the bioavailability and toxicity of NACs during bioremedation. We have measured the toxicity of 2,4-dinitrotoluene (2,4-DNT) to duckweed grown in smectite suspensions and utilized Ca2+/K+ exchange to retain or release 2,4-DNT. Retention by potassium smectite reduced bioavailability and hence toxicity to duckweed. Addition of Ca2+ to replace K+ by ion exchange released adsorbed 2,4-DNT, which is toxic to duckweed. So smectites can be used to sequester or release 2,4-DNT predictably and provide means to control bioavailability and environmental toxicity. PMID- 17396655 TI - Electronic polarizability as a predictor of biodegradation rates of dimethylnaphthalenes. an ab initio and density functional theory study. AB - Geometries, relative stabilities, electronic excited states, atomic charges, and electronic dipole polarizabilities of dimethylnaphthalene (DMN) isomers have been calculated in gas and aqueous phases by ab initio and DFT methods. At the highest levels of calculation, alpha,alpha-DMN (2,6-DMN, 2,7-DMN, and 2,3-DMN) are the lowest energy isomers, while 1,8-DMN is the less stable by 7-8 kcal mol(-1). The averaged electronic polarizability, , is dependent on the position of the methyl substituents, increasing in the order alpha, alpha-DMN < (a, beta-DMN < beta, beta-DMN, with the largest values being obtained for 2,6-DMN and 2,7-DMN, while the lowest value is calculated for 1,8-DMN isomer. Polarizability differences among the isomers have been related to their spectroscopic properties. The computed value of DMN isomers, with the notable exception of 2,7-DMN, is in excellent linear relationship with the observed first-order biomass-normalized rate coefficient, a parameter related to the rate of biodegradation of polycyclic aromatic hydrocarbons (PAHs). This result suggests that electronic polarizability may be a useful tool for prediction of biodegradation trends of series of compounds, and inductive and dispersive interactions play a fundamental role in the biodegradation process of DMNs. The present approach is potentially suitable for applications on PAHs with higher molecular weight. PMID- 17396656 TI - Multivariate chemical mapping of antibiotics and identification of structurally representative substances. AB - Antibiotics used in human and veterinary medicine have been found in samples from diverse environments in many parts of the world. To assess the environmental risks associated with them, data regarding their toxicity, occurrence, and fate are needed, but gathering such data is time-consuming and expensive. An efficient approach to address these difficulties would be to select a small subset of antibiotics with a wide variation in chemical characteristics, perform experimental tests on this subset, and then extrapolate the results to larger numbers of antibiotics, including the most potentially hazardous compounds. To assess the potential utility of such an approach, a set of 92 antibiotics for human use was studied and their structural properties were described with 24 chemical descriptors that included information on their steric, lipophilic, and electronic properties. Principal component analysis in combination with statistical experimental design was used to map the chemical diversity of the antibiotics and to select a small subset, a "training set", of 20 antibiotics. The chemical representativity of the training set was assessed in a quantitative structure-activity model established to predict ultimate biodegradation. The selected antibiotics showed to cover the chemical variation of the studied antibiotics and are suggested for use in future testing programs to assess antibiotics' fate and effects in the environment. PMID- 17396657 TI - Volatile organic compounds in human milk: methods and measurements. AB - The present study was conducted to optimize methods for measurement of volatile organic compounds (VOCs) by use of headspace solid-phase microextraction (HS SPME) and to provide a preliminary assessment of levels in human milk. MTBE (methyl tert-butyl ether), chloroform, benzene, and toluene were measured from two sources of milk: a North Carolina milk bank (n = 5) and multiple samples from three women within nonsmoking households in inner-city Baltimore, MD (n = 8). In Baltimore, indoor air VOC concentrations in the respective households were also measured by active sampling and thermal desorption gas chromatography/mass spectrometry in selective ion monitoring (GC/MS/SIM) over each of the 3 days of milk collection. By application of these optimized methods, we observed median VOC concentrations in Baltimore human milk of 0.09, 0.55, 0.12, and 0.46 ng/mL for MTBE, chloroform, benzene, and toluene, respectively. For benzene, toluene, and MTBE, milk levels trended with observed indoor air concentrations. On the basis of measured concentrations in air and milk, infant average daily dose by inhalation exceeded ingestion rates by 25-135-fold. Thus, VOC exposure from breast milk is vastly exceeded by that from indoor air in nonsmoking households. Accordingly, strategies to mitigate infant VOC exposure should focus on the indoor air inhalation pathway of exposure. PMID- 17396658 TI - A method of measuring Escherichia coli 0157:H7 at 1 cell/mL in 1 liter sample using antibody functionalized piezoelectric-excited millimeter-sized cantilever sensor. AB - Piezoelectric-excited millimeter-sized cantilever (PEMC) sensors immobilized with antibody specific to Escherichia coli (EC) 0157:H7 is used to detect EC at 1 cell/mL in 1 mL and 1 L samples in a batch and flow mode, respectively. Two sensor designs were used. The first design (PEMC-a) has both the piezoelectric and non-piezoelectric layer anchored, while in the second design (PEMC-b) had only the piezoelectric layer anchored. PEMC-a, used in batch mode with 1 mL sample, showed limit of detection at 10 cells/ mL using the second bending mode at 85.5 kHz in air. PEMC-b exhibited resonant frequencies at 186.5, 883.5, and 1778.5 kHz in air and 162.5, 800.0, and 1725.5 kHz in sample flow conditions. A one-liter sample containing 1000 EC cells was introduced at 1.5, 2.5, 3, and 17 ml/min, and the change in resonant frequency was monitored. The total frequency change observed for the mode at 800 kHz and sample flow rates of 1.5, 2.5, 3, and 17 mL/min were 2230+/-11, 3069 +/-47, 4686+/-97, and 7188+/-52 Hz, respectively. Each detection experiment was confirmed by exposing the sensor to a low pH solution followed by a phosphate buffered saline (PBS) rinse, which caused the release of the attached EC. The final frequency change observed was nearly identical to the value prior to EC attachment. Kinetic analysis showed that the observed binding rate constant at 1.5, 2.5, 3 mL/min were 0.009, 0.015, and 0.021 min(-1), respectively. The significance of these results is that very low concentration of pathogens in large sample volumes can be measured in a short time period without the need for filtration or enrichment. PMID- 17396659 TI - Causes of variable biomass density and its effects on settleability in full-scale biological wastewater treatment systems. AB - Both variable biomass density and floc structure were determined to affect the settleability of microbial biomass produced during biological wastewater treatment (activated sludge). Average biomass density varied from 1.022 to 1.056 g/mL in a survey of 17 full-scale biological wastewater treatment systems with a variety of configurations. Biomass settleability was correlated with density in samples with higher filament contents and/or more open floc structures, but settleability was independent of density in biomass with lower filament contents and more rounded floc structures. Biomass density increased with polyphosphate content, and enhanced biological phosphorus removal (EBPR) plants had higher density and better settleability than non-EBPR plants, including two systems that converted to EBPR during the course of this study. Density also increased with increasing nonvolatile suspended solids content, which was linked both to polyphosphate and to increasing solids residence times. Both density and floc structure should be considered in future analyses of activated sludge settleability, and it may be possible to improve system performance by adopting a new set of operational and design strategies to increase density. PMID- 17396660 TI - UV degradation kinetics and modeling of pharmaceutical compounds in laboratory grade and surface water via direct and indirect photolysis at 254 nm. AB - Direct and indirect photolysis of pharmaceutically active compounds (PhACs) was investigated in laboratory-grade water (LGW) and a local surface water using a low-pressure ultraviolet batch reactor. The PhACs selected in this study belong to different therapeutic classes and are known to occur in environmental samples. Fundamental photolysis and advanced oxidation process parameters obtained in LGW (such as the decadic molar absorption coefficient, quantum yield, and degradation rate constants) are reported and discussed. These parameters, together with the incident photon irradiance, solution depth, and solution absorbance were used to develop UV and UV/ H202 photolysis models that were compared with experimental results obtained in the surface water. The model predicted the experimental UV photolysis removals well but underestimated the UV/H2O2 photolysis results. These models were used to discuss the effects of optical path length and H2O2 concentration on the UV-based rate constant predictions. PMID- 17396661 TI - Study of an axial flow cyclone to remove nanoparticles in vacuum. AB - An axial flow cyclone for removing nanoparticles was tested for collection efficiency. Data were validated numerically in vacuum conditions of several Torrs, with flow rates of 0.35-0.57 slpm. The experimental cutoff aerodynamic diameter of the cyclone ranged from 21.7 to 49.8 nm. A 3-D numerical simulation was conducted first to calculate detailed flow and pressure fields, then a Brownian Dynamics simulation was done to determine the collection efficiency of nanoparticles. Both centrifugal force and Brownian diffusion were taken into account. The simulated results for both pressure drop and cutoff aerodynamic diameter are in good agreement with the experimental data. In comparison, previous theories using simplified tangential flow field assumption are not able to predict collection efficiencies accurately. The numerical model developed in this study can facilitate cyclone design to classify valuable nanopowders below a certain diameter, or to remove toxic nanoparticles from the vacuum exhaust of process chambers commonly used in high-tech industries. PMID- 17396662 TI - Trichloroethene degradation by UV/H2O2 advanced oxidation process: product study and kinetic modeling. AB - The broadband UV irradiation of 1.1 mM trichloroethene (TCE) aqueous solution in the presence of 10.4 mM H2O2 resulted in formic, oxalic, dichloroacetic (DCA), and monochloroacetic (MCA) acids, as organic byproducts. The organic chlorine was converted completely to chloride ion as a final product. TCE and its degradation products were completely mineralized in 30 min, under a volume-averaged UV-C irradiant power of 35.7 W/L from a 1 kW medium-pressure mercury vapor arc lamp. TCE degraded primarily through hydroxyl radical-induced reactions and onlyto a low extentthrough direct UV photolysis and chlorine atom-induced chain reactions. The experimental patterns of TCE, H2O2, and detected reaction products combined with the literature information on radical reactions in the aqueous phase were used to postulate a degradation mechanism and to develop a kinetic model to predict the TCE decay, formation and degradation of byproducts, and pH and oxygen profiles. The agreement between the model calculations and the experimental data is satisfactory. PMID- 17396663 TI - Decomposition of N20 over hexaaluminate catalysts. AB - Catalytic N2O decomposition has been studied over metal-substituted hexaaluminates with the general formula ABAl11O19, where A = La, Ba, and B = Mn, Fe, Ni. The materials were prepared by coprecipitation via the carbonate route followed by calcination at 1473 K for 10 h. Inductively coupled plasma optical emission spectroscopy (ICP-OES), X-ray diffraction (XRD), transmission electron microscopy (TEM), and N2 adsorption techniques were used to characterize the catalysts. The activity in direct N20 decomposition was evaluated by means of temperature-programmed reaction and steady-state experiments. Fe- and Mn containing hexaaluminates present the highest activities. The Ni-containing catalysts are significantly less active, comparable to the hexaaluminates without metal substitution. The catalytic activity was practically not influenced by the A cation (La or Ba) in the structure. The Fe- and Mn-substituted hexaaluminates exhibit high activity and stability for N2O decomposition in mixtures simulating the outlet of the Pt-Rh gauzes in ammonia oxidation reactors, containing N2O, NO, O2, and H2O. These materials are promising for high-temperature abatement of this powerful greenhouse gas in the chemical industry, particularly in nitric acid and caprolactam production. PMID- 17396664 TI - Effects of the nonionic surfactant tween 80 on microbial reductive dechlorination of chlorinated ethenes. AB - Recent field studies have indicated synergistic effects of coupling microbial reductive dechlorination with physicochemical remediation (e.g., surfactant flushing) of dense nonaqueous phase liquid (DNAPL) source zones. This study explored chlorinated ethene (e.g., tetrachloroethene [PCE]) dechlorination in the presence of 50-5000 mg/L Tween 80, a nonionic surfactant employed in source zone remediation. Tween 80 did not inhibit dechlorination by four pure PCE-to-cis-1,2 dichloroethene (cis-DCE) or PCE-to-trichloroethene (TCE) dechlorinating cultures. In contrast, cis-DCE-dechlorinating Dehalococcoides isolates (strain BAV1 and strain FL2) failed to dechlorinate in the presence of Tween 80. Bio-Dechlor INOCULUM (BDI), a PCE-to-ethene dechlorinating consortium, produced cis-DCE in the presence of Tween 80, further suggesting that Tween 80 inhibits dechlorination by Dehalococcoides organisms. Quantitative real-time PCR analysis applied to BDI revealed that the number of Dehalococcoides cells decayed exponentially (R(2) = 0.85) according to the Chick-Watson disinfection model (pseudo first-order decay rate of 0.13+/-0.02 day(-1)) from an initial value of 6.6 +/-1.5 x 10(8) to 1.3+/-0.8 x 10(5) per mL of culture after 58 days of exposure to 250 mg/L Tween 80. Although Tween 80 exposure prevented ethene formation and reduced Dehalococcoides cell numbers, Dehalococcoides organisms remained viable, and dechlorination activity pist cis-DCE was recovered following the removal of Tween 80. These findings suggest that sequential Tween 80 flushing followed by microbial reductive dechlorination is a promising strategy for remediation of chlorinated ethene-impacted source zones. PMID- 17396665 TI - Formation and durability of dithiocarbamic metals in stabilized air pollution control residue from municipal solid waste incineration and melting processes. AB - In Japan, chemical stabilization of metals using a chelating agent composed of dithiocarbamic salt is the prevailing technique for the final disposal of air pollution control (APC) residue from municipal solid waste (MSW) incineration and melting processes. The objective of this study is to quantitatively evaluate the formation and durability of dithiocarbamic metals (DCM) in treated APC residue. In this research, sodium N,N--diethyldithiocarbamate trihydrate was used as a chelating agent. An extraction method using n-butyl acetate was developed to separate DCM from residue. This method elucidated the competitive formation of DCM among Cu, Pb, Cd, and Zn. The metal fraction forming DCM was about 70 and 90% of the total content of Pb in APC residue from incineration and melting, respectively. Regarding durability in the environment, 240 day curing of the chelate-treated APC residue showed that air-contact conditions induced degradation of DCM with time. On the other hand, sealed conditions did not bring about a decrease over the course of the experiment. These results imply that DCM degradation proceeds when the treated residue comes into contact with air in actual landfills. PMID- 17396666 TI - Photodegradation of rhodamine B in water assisted by titania nanorod thin films subjected to various thermal treatments. AB - Well-crystallized titania nanorod thin films with high specific surface areas were synthesized through in situ oxidation of metallic titanium substrates with hydrogen peroxide solutions at a low temperature of 80 degrees C. The effects of thermal treatments on the ability of the thin films to assist photodegradation of rhodamine B in water were studied in detail. It is found that, due to a unique nanofeature, the titania nanorod film retained its small grain size and high specific surface area during the subsequent thermal treatment for up to 450 degrees C, which further improved the crystallinity of titania. In addition, upon heating to 450 degrees C, the nanorods still possessed a high surface fractal dimension, which is an indication of high surface roughness and surface area. As a result, the photocatalytic activity of the titania nanorod film is found to increase with an increasing heating temperature for up to 450 degrees C. The subsequent thermal treatment at temperatures beyond 450 degrees C decreased the photocatalytic activity, because of the significantly reduced specific surface area. The current investigation provided a simple and easily scaled-up approach to produce photocatalysts for efficient removal of dye effluents in wastewater. PMID- 17396667 TI - Distribution and abiotic degradation of chlorinated solvents in heated field samples. AB - The objective of this study was to evaluate the abiotic degradation of tetrachloroethylene (PCE) in contaminated soil and groundwater samples obtained from the Camelot Cleaners Superfund site, West Fargo, ND. The field samples were incubated at temperatures of 25, 55, 75, and 95 degrees C in sealed ampules containing aqueous, gas, and solid phases for periods of up to 75 days to simulate thermal treatment temperatures. Aqueous PCE concentrations increased with incubation temperature but remained constant over time. The degradation of dolomite to form CO2 facilitated the transfer of sorbed-phase PCE from the solid to the aqueous phase in heated ampules. While compounds associated with PCE degradation were detected in the heated ampules, these compounds were also detected in ampules with PCE-free Camelot soil and were attributed to soil diagenesis rather than PCE degradation. Trichloroethylene underwent hydrogenolysis to form cis-DCE at 95 degrees C, and TCE levels decreased with first-order half-lives of 157 days at 55 degrees C and 26 days at 95 degrees C. The relatively small decrease in total PCE levels after 75 days of heating at 95 degrees C suggests that abiotic degradation of PCE will not result in significant mass reduction during thermal treatment of the Camelot Cleaners Superfund site. PMID- 17396668 TI - Novel mercury control technology for solid waste incineration: sodium tetrasulfide (STS) as mercury capturing agent. AB - Traditional pollution control technologies are able to capture oxidized forms of mercury to some extent; however, they show low efficiency for the control of elemental mercury emissions. This study developed a novel mercury removal technology: injection of sodium tetrasulfide (Na2S4) dissolved in the sodium hydroxide (NaOH) solution in the spray-dryer system. The effects of flue gas temperature and Na2S4 level in flue gas on the mercury removal efficiency were investigated. Na2S4 was decomposed into Na2S (S2-) and elemental S (S0), which reacted with HgCl2 and elemental Hg (Hg0), and HgS was then formed. Under the optimized operation parameters, this technology can simultaneously remove over 88% of HgCl2 and more than 90% of Hg0 from a flue gas stream containing about 400 microg m(-3) Hg0 and 1200 microg m(-3) HgCl2. The increased flue gas temperature (>170 degrees C) and the decreased Na2S4-to-Hg mass ratio (S-Hg-R) (<2.0) had negative effects on the reactions of gaseous mercury (HgCl2 + Hg0) with ionic sulfur (S2-) and S0. All the experiments were conducted in a full scale hospital waste incinerator with a capability of 20 tons per day (TPD). PMID- 17396669 TI - Electrochemical stimulation of microbial perchlorate reduction. AB - As part of our studies into the diversity of dissimilatory perchlorate reducing bacteria (DPRB) we investigated the reduction of perchlorate in the cathodic chamber of a bioelectrical reactor (BER). Our results demonstrated that washed cells of Dechloromonas and Azospira species readily reduced 90 mg L(-1) perchlorate in the BER with 2,6-anthraquinone disulfonate (AQDS) as a mediator. No perchlorate was reduced in the absence of cells or AQDS, or in an open-circuit control. Similar results were observed when a natural microbial community was inoculated into a fed-batch BER. After 70 days of operation, a novel DPRB, strain VDY, was isolated which readily reduced perchlorate in a mediatorless BER. Continuous up-flow BERs (UFBERs) were seeded with active cultures of strain VDY, and perchlorate at a volumetric loading of 60 mg L(-1) day(-1) was successfully removed. Gas phase analysis indicated that low levels of H2 produced at the cathode surface through electrolysis may mediate this metabolism. The results of these studies demonstrate that biological perchlorate remediation can be facilitated through the use of a cathode as the primary electron donor, and that continuous treatment in such a system approaches current industry standards. This has important implications for the continuous treatment of this critical contaminant in industrial waste streams and drinking water. PMID- 17396670 TI - Rapid response concentration-controlled desorption of activated carbon to dampen concentration fluctuations. AB - Fluctuations in concentration of organic vapors in gas streams that are treated by devices such as biofilters or oxidizers make it challenging to remove the vapors from the gas streams in an efficient and economic manner. Combining adsorption with concentration-controlled desorption provides an active buffer between the source of vapors and the control device for better control of concentration and flow rate of the gas stream that is treated by the secondary control device, hence further enhancing the performance or reducing the size of the devices. Activated carbon fiber cloth is used with microwave swing adsorption to remove methyl ethyl ketone (MEK) from air streams and then provide a readily controllable feed stream of that vapor in air at a specified concentration and gas flow rate with steady-state tracking desorption. MEK was captured with >99.8% efficiency during the adsorption cycle. The MEK concentration during the regeneration cycle was readily controlled at concentration set-points between 170 and 5000 ppmv, within relative standard deviations of 1.8 and 4.9%, respectively, and at 20% of the gas flow rate that was treated during the adsorption cycle. Such capability of the system allows the secondary control device to be optimized for select constant concentrations and low gas flow rates that is not possible without such pretreatment. PMID- 17396671 TI - Selenium toxicity to invertebrates: will proposed thresholds for toxicity to fish and birds also protect their prey? AB - Efforts to manage the environmental risks of selenium (Se) in freshwater ecosystems have focused primarily on fish and birds, with invertebrates most often considered only as dietary sources of Se to higher trophic levels. Relatively little attention has been given to the risk of Se toxicity to invertebrates. Based on a review of 156 aqueous, dietary, or internal Se concentrations associated with toxic effects in 29 macroinvertebrate species, we found that water concentrations associated with acute lethality varied > 1000 fold among taxa, whereas toxic dietary concentrations varied approximately 100 fold and toxic internal concentrations varied about 30-fold. Sublethal effects occurred at approximately 10-fold lower concentrations than lethality. Sublethal effects occurred at 1-30 microg Se/g dry weight in invertebrate tissue, a range that encompasses proposed dietary thresholds for toxicity to fish and water birds, suggesting that Se may cause toxic effects in some invertebrate species at concentrations considered to be "safe" for the organisms consuming them. PMID- 17396672 TI - Effects of chytrid and carbaryl exposure on survival, growth and skin peptide defenses in foothill yellow-legged frogs. AB - Environmental contaminants and disease may synergistically contribute to amphibian population declines. Sub-lethal levels of contaminants can suppress amphibian immune defenses and, thereby, may facilitate disease outbreaks. We conducted laboratory experiments on newly metamorphosed foothill yellow-legged frogs (Rana boylii) to determine whether sublethal exposure to the pesticide carbaryl would increase susceptibility to the pathogenic chytrid fungus Batrachochytrium dendrobatidis that is widely associated with amphibian declines. We examined the effect of carbaryl alone, chytrid alone, and interactions of the two on individual survival, growth, and antimicrobial skin defenses. We found no effect of chytrid, carbaryl, or their interaction on survival. However, chytrid infection reduced growth by approximately one-half. This is the first report of suppressed growth in post-metamorphic amphibians due to infection with chytrid. Rana boylii skin peptides strongly inhibited chytrid growth in vitro, which may explain why chytrid exposure did not result in significant mortality. Skin peptide defenses were significantly reduced after exposure to carbaryl suggesting that pesticides may inhibit this innate immune defense and increase susceptibility to disease. PMID- 17396673 TI - Predicting copper toxicity with its intracellular or subcellular concentration and the thiol synthesis in a marine diatom. AB - The accumulation and subcellular distribution of copper (Cu) in a marine diatom Thalassiosira weissflogii were examined under different nutrient conditions [nitrogen- (-N) and phosphorus-starved (-P) conditions as well as nutrient enriched conditions with nitrate (+NO3-) and ammonium (+NH4+) as the nitrogen source]. Cu accumulation was induced in the NO3--exposed (+N03- and -P) cells, suggesting that Cu may be directly or indirectly required for intracellular NO3- reduction. The relationships between the response of the cell-specific growth rate or photosynthetic system II maximum quantum yield ((DM) to Cu exposure in different nutrient-conditioned cells and the free copper ion concentration,the intracellular Cu concentration (intra-Cu), and the distribution of Cu in different subcellular compartments were also examined. We found that the intra-Cu was the best Cu toxicity predictor, as it accounted for most of the Cu-induced PhiM response between different nutrient-conditioned cells. The synthesis of intracellular cysteine and five low molecular weight (LMW) thiols was not notably induced at high Cu levels possibly because of the existence of some other Cu detoxification mechanisms. This was further demonstrated by the much lower cysteine and LMW thiol contents in the -N cells with a similar Cu tolerance to the nutrient-enriched cells. Overall, our results suggest that Cu toxicity could be better predicted with the intra-Cu and its subcellular distribution as compared with the currently used free ion activity model and biotic ligand model. However, the LMW thiols had unexpectedly little contribution to Cu detoxification in T. weissflogii. PMID- 17396674 TI - Variation in the bioaccumulation of a sediment-sorbed hydrophobic compound by benthic macroinvertebrates: patterns and mechanisms. AB - Aquatic ecological risk assessment is primarily focused on aqueous exposure, but many hydrophobic contaminants bind to particulate material and accumulate in sediments. The risk posed by such contaminants is partially dependent on the importance of dietary exposure. Here, we describe the bioaccumulation of a highly hydrophobic compound (dioctadecyl-dimethyl ammonium chloride (DODMAC)) to four freshwater macroinvertebrates (i.e., Asellus aquaticus, Chironomus riparius, Gammarus pulex, Lumbriculus variegatus) and investigate the mechanistic basis for observed interspecific variation in bioaccumulation. Although more than 99.99% of DODMAC was sediment-bound, it was bioavailable to all four species via dietary exposure. Interspecific variation in bioaccumulation was apparent despite the lack of selective feeding and biotransformation potential and after normalization for body size and lipid content. Chironomus riparius had the highest lipid normalized DODMAC concentration and L. variegatus had the lowest. Study species differed in factors affecting uptake (i.e., feeding rate) and absorption efficiency (i.e., gut passage time and gut surfactancy). Feeding rate did not explain interspecific variation in bioaccumulation, but bioaccumulation was enhanced by either high surfactancy and short gut passage time (e.g., G. pulex) or low surfactancy and long gut passage time (e.g., C. riparius). Risk assessment of hydrophobic contaminants should consider dietary exposure and the potential food chain effects of interspecific variation in bioaccumulation. PMID- 17396675 TI - Prokaryotic gene profiling assays to detect sediment toxicity: evaluating the ecotoxicological relevance of a cell-based assay. AB - Despite their complexity, ecotoxicological measurements using higher level responses remain a major tool in the assessment of ecosystem integrity. Nevertheless, the past decade saw an increasing number of cell based testing systems have found widespread application in ecotoxicology. One such test is bacterial bioreporters carrying a stress sensitive promoter fused to an easily detectable reporter gene. In the presence of a specific toxic stress,the expression cassette is switched on and the reporter gene is produced. This study evaluated the use of 14 different Escherichia coli bioreporter strains sensitive to different types of toxicity in the assessment of the ecological status of a small river basin in Flanders, Belgium. The river is fed at two geographically separate locations by two distinct and well-described effluents, one from a household sewage treatment facility and one from the discharge of the wastewater treatment facility of a large chemical plant. The results of the bacterial gene profiling assay were related to active biomonitoring results obtained through higher-level responses of caged Dreissena polymorpha, Chironomus riparius, and Cyprinus carpio deployed at the locations sampled for the bacterial assay. The results of the gene induction assay and the active biomonitoring data correlated well and corresponded to the flow dilution data, which is used here as a surrogate forthe chemical pollution gradient present in the river basin. PMID- 17396676 TI - Comment on "Discrimination of shifts in a soil microbial community associated with TNT-contamination using a functional ANOVA of 16S rRNA hybridized to oligonucleotide microarrays". PMID- 17396677 TI - Point and click. PMID- 17396678 TI - Orient express. Ease navigation of your children's hospital by creating a patient/visitor's guide. PMID- 17396679 TI - Hire drill. Competent culture hiring isn't what you think. PMID- 17396680 TI - Built to last. Undertake market research before embarking on facility expansion or renovation. PMID- 17396681 TI - Round bounty. One-hour rounding positively influences patients and nursing staff members. PMID- 17396682 TI - Total recall. Improve consumer satisfaction and recall of healthcare information with social learning theory. PMID- 17396683 TI - Join the evolution. Has traditional marketing met its match with online social networking? PMID- 17396685 TI - Do your research. PMID- 17396684 TI - Speak up. PMID- 17396686 TI - Reversal of fortune. PMID- 17396687 TI - Panretinal photocoagulation and photodynamic therapy for anterior segment neovascularization secondary to ischemic central retinal vein occlusion. AB - BACKGROUND AND OBJECTIVE: To compare the effects of panretinal photocoagulation (PRP) and photodynamic therapy (PDT) for anterior segment neovascularization secondary to ischemic central retinal vein occlusion (CRVO). PATIENTS AND METHODS: Fifty-seven eyes were randomized to receive standard PRP (19 eyes), selective PRP (20 eyes), or PDT (17 eyes). Selective PRP was performed only when iris neovascularization and/or angle neovascularization showed progression on weekly follow-up. Primary outcome measures were the extension of anterior segment neovascularization and the rate of neovascular glaucoma development. Secondary outcome measures included visual acuity results. RESULTS: After 12 months of follow-up, iris neovascularization extension was 0.52, 2.55, and 2.27 in the PRP, selective PRP, and PDT groups, respectively. Anterior segment neovascularization extension was 0.57, 1.50, and 1.27 in the PRP, selective PRP, and PDT groups, respectively. Both showed a statistically significant difference in the PRP group. The rate of neovascular glaucoma development was similar in the three groups. CONCLUSIONS: Although PRP was better at determining iris neovascularization and anterior segment neovascularization regression, the similar rate of neovascular glaucoma development found in the three groups indicates that anterior segment neovascularization secondary to ischemic CRVO can also be safely managed using selective PRP or PDT. PMID- 17396688 TI - Incidence of complications in 25-gauge transconjunctival sutureless vitrectomy based on the surgical indications. AB - BACKGROUND AND OBJECTIVE: The complications of 25-gauge transconjunctival sutureless vitrectomy based on the surgical indications were reviewed. PATIENTS AND METHODS: Thirty-eight patients underwent 25-gauge transconjunctival sutureless vitrectomy. Indications for vitrectomy, preoperative and postoperative best-corrected visual acuity, preoperative and postoperative intraocular pressure, and any intraoperative or postoperative complications were recorded. RESULTS: Complications included the need to suture a leaking sclerotomy intraoperatively in four eyes (9%), all of which involved oil removal; postoperative hypotony with choroidals in two eyes (5%); mild progression of nuclear sclerotic cataract in one eye (2% of all eyes, 4% of phakic eyes); the need to switch to 20-gauge pars plana vitrectomy in one eye (2%); corneal abrasion in one eye (2%); and retinal detachment in one eye (2%). Sclerotomy leakage and hyopotony with choroidals were only encountered in cases involving previously vitrectomized eyes, whereas none of the eyes without previous vitrectomy had leakage-related complications. CONCLUSIONS: Previously vitrectomized eyes have a higher incidence of complications related to postoperative leakage, possibly due to the lack of plugging effect of peripheral vitreous on the unsutured sclerotomy. One should consider suturing any leaking sclerotomy at the conclusion of surgery involving previously vitrectomized eyes. PMID- 17396689 TI - A retrospective study of the incidence of culture-positive endophthalmitis after cataract surgery and the use of preoperative antibiotics. AB - BACKGROUND AND OBJECTIVE: To determine the incidence of culture-positive endophthalmitis after cataract surgery in an ambulatory surgery center and to analyze the effect of a specific preoperative antibiotic regimen in reducing the incidence of endophthalmitis. PATIENTS AND METHODS: The medical records of 30,870 consecutive eyes undergoing cataract extraction at a single ambulatory surgery center were reviewed retrospectively. Complications of the cataract surgeries, including endophthalmitis, were collected. RESULTS: Cataract surgery was performed without the use of preoperative topical antibiotics on 19,529 eyes from 1990 to 1998 (Group 1) and using topical preoperative ciprofloxacin-soaked pledgets on 11,341 eyes from 1999 to 2003 (Group 2). Five eyes had culture positive endophthalmitis in each group. The incidence of culture-positive endophthalmitis was 0.03% (1 in 3000) in Group 1 and 0.04% (1 in 2268) in Group 2 (P = 1.0). CONCLUSIONS: The incidence of postoperative endophthalmitis in this study was low. A preoperative pledget soaked with a second-generation fluoroquinolone did not appear to lower the incidence of culture-positive endophthalmitis after cataract surgery in this series. PMID- 17396690 TI - A survey of desired skills to acquire in ophthalmology training: a descriptive statistical analysis. AB - BACKGROUND AND OBJECTIVE: To determine which skills are most important to clinicians and how they correlate with Accreditation Council for Graduate Medical Education (ACGME) competencies. PATIENTS AND METHODS: A descriptive analysis to test four hypotheses: specific skill sets are likely to be important/mandatory for residents to obtain during training; comprehensive and subspecialist ophthalmologists would likely value skills similarly; year of training would likely affect the ranking of skills; and surveying practicing ophthalmologist could help to modify the educational curriculum. RESULTS: Highest-ranked skill sets included interpersonal and communication skills (mean, 1.17 +/- 0.08), ocular trauma (mean, 1.28 +/- 0.26), and practice-based learning (mean, 1.37). Comprehensive ophthalmologists and subspecialists did not significantly differ in the ranking of skill sets. Year of graduation did not affect the ranking of skill sets. CONCLUSION: The ACGME core competencies of interpersonal and communication skills and practice-based learning were ranked as highly important to both comprehensive and subspecialty ophthalmologists. These are included in ACGME core competency requirements. These findings support ACGME core competencies and the need for continued evaluation of ophthalmology residents in all areas of medical training. PMID- 17396691 TI - Sonographic features of senile scleral calcification. AB - BACKGROUND AND OBJECTIVE: To describe the sonographic features of senile scleral calcification using ultrasound biomicroscopy. PATIENTS AND METHODS: Four patients in whom senile scleral calcifications were found incidentally on computed tomography studies underwent ultrasound biomicroscopy examination. RESULTS: The senile scleral calcification was bilateral in three patients and unilateral in one patient. In all patients, the calcified material created the anterior shadowing typical of calcified lesions. The ultrasound biomicroscopy study localized the senile scleral calcification to the subconjunctival layer, superficially above the sclera, and anterior to the horizontal muscles. CONCLUSIONS: This is the first description of the ultrasound biomicroscopy features of senile scleral calcification. Ultrasound biomicroscopy may serve as a good tool for the accurate localization of the plaque. PMID- 17396692 TI - Retinal nerve fiber layer thickness evaluation using optical coherence tomography in eyes with optic disc hemorrhage. AB - BACKGROUND AND OBJECTIVE: To examine the association between disc hemorrhage and retinal nerve fiber layer (RNFL) thickness. PATIENTS AND METHODS: RNFL thicknesses were measured using optical coherence tomography at the location of disc hemorrhage, at the superior or inferior symmetrical area of the same eyes, and at equivalent locations in contralateral eyes without disc hemorrhage. RNFL thicknesses were compared using the paired-sample t test. For recurrent hemorrhages, the correlation between the number of recurrences and RNFL thickness was examined using Kendall's tau-b method. RESULTS: Thirty-two disc hemorrhages were found in 32 eyes with normal tension or primary open-angle glaucoma. RNFL thicknesses in areas with disc hemorrhage were thinner than those in superior or inferior symmetrical locations in the same eyes (81.7 +/- 32.9 vs 100.7 +/- 34.5 microns, P = .005), but were not different from those in equivalent locations in contralateral eyes (92.8 +/- 31.7 microns, P = .092). Number of recurrent hemorrhages and RNFL thicknesses at sites of recurrent hemorrhages tended to show a negative correlation (correlation coefficient = -0.260, P = .079). CONCLUSIONS: Disc hemorrhage is associated with RNFL thinning measured by optical coherence tomography, and a negative correlation was found between recurrent hemorrhages and RNFL thickness. PMID- 17396693 TI - Computerized detection and measurement of drusen in age-related macular degeneration. AB - BACKGROUND AND OBJECTIVE: The authors used a drusen detection algorithm to quantitate drusen on digitized images and determined its precision and accuracy. PATIENTS AND METHODS: Fundus images from 349 participants in the Age-Related Eye Disease Study trial were digitized and analyzed with the Drusen Analyzer (IRIDEX Corporation, Mountain View, CA). The size, number, and area of the drusen in two macular regions were computed by two readers using an interactive approach. Measurements were compared to data generated by reading center methods. RESULTS: For the Analyzer, inter-observer agreement was 0.79 (SE = 0.02) and 0.86 (SE = 0.01) (kappa statistic). Intra-observer precision was 0.835 and 0.880 (Spearman coefficients). Categorical agreement (weighted kappa) between the Analyzer and reading center results for the two regions was (0.76, 0.58) and (0.68, 0.68) for readers A and B. The time required to quantitate an image using the Analyzer was 105 +/- 52 and 218 +/- 102 seconds for the two regions, respectively. CONCLUSIONS: Quantitative detection of drusen can be performed reproducibly and efficiently. Comparisons to more labor intensive reading center techniques suggest that the results are similar but that the algorithm is more sensitive and precise. PMID- 17396694 TI - Corneal edema and permanent blue discoloration of a silicone intraocular lens by methylene blue. AB - BACKGROUND AND OBJECTIVE: To report a silicone intraocular lens (IOL) stained blue by inadvertent intraoperative use of methylene blue instead of trypan blue and the results of experimental staining of various lens materials with different concentrations of the same dye. MATERIALS AND METHODS: A "blue dye" was used to enhance visualization during capsulorhexis in a patient undergoing phacoemulsification with implantation of a three-piece silicone lens. Postoperatively, the patient presented with corneal edema and a discolored IOL. Various IOL materials were experimentally stained using methylene blue. Sixteen lenses (4 silicone, 4 hydrophobic acrylic, 4 hydrophilic acrylic, and 4 polymethylmethacrylate) were immersed in 0.5 mL of methylene blue at concentrations of 1%, 0.1%, 0.01%, and 0.001%. These lenses were grossly and microscopically evaluated for discoloration 6 and 24 hours after immersion. RESULTS: The corneal edema resolved within 1 month after the initial surgical procedure. After explantation, gross and microscopic analyses of the explanted silicone lens revealed that its surface and internal substance had been permanently stained blue. In the experimental study, all of the lenses except the polymethylmethacrylate lenses were permanently stained by methylene blue. The hydrophilic acrylic lenses showed the most intense blue staining in all dye concentrations. CONCLUSIONS: This is the first clinicopathological report of IOL discoloration due to intraocular use of methylene blue. This and other tissue dyes may be commonly found among surgical supplies in the operating room and due diligence is necessary to avoid mistaking these dyes for those commonly used during ocular surgery. PMID- 17396695 TI - Intraocular biodistribution of mono-L-aspartyl chlorin e6 in a primate choroidal neovascularization model. AB - BACKGROUND AND OBJECTIVE: To describe the biodistribution of a hydrophilic sensitizer, mono-L-aspartyl chlorin e6 (NPe6), in a primate model of choroidal neovascularization (CNV). MATERIALS AND METHODS: NPe6 (25 mg/kg) and indocyanine green (ICG, 15 mg/kg) were intravenously administered simultaneously in monkeys with laser-induced CNVs. Eyes were enucleated and examined by fluorescence microscopy immediately, 20 minutes, and 1, 4, and 24 hours after dye injection. RESULTS: Fluorescence peaked in retinal vessels immediately after dye administration; rapid washout began 1 hour postinjection. In contrast, experimental CNV demonstrated little fluorescence immediately after injection, with increased intensity at later time points. Peak dye fluorescence in CNV occurred 1 hour following dye injection. NPe6 fluorescence diminished rapidly and washed out completely by 24 hours. The dye accumulation and retention pattern of ICG dye resembled that of NPe6. CONCLUSIONS: This study demonstrated the selective accumulation of NPe6 photosensitizer and ICG dye within experimental CNV and rapid clearance from the circulation and retinochoroidal tissue. PMID- 17396696 TI - Clinical anophthalmia with orbital heterotopic brain tissue. AB - The authors describe the uncommon clinical presentation and treatment of a neonate with progressive proptosis, which turned out to be an ectopic cerebral rest in the orbit in the absence of a formed eye. The discussion examines the theories regarding the genesis of the condition, different locations in the body (including the orbit and the eye) in which ectopic cerebral rests have been described in the literature, and the management of the condition. PMID- 17396697 TI - Clinical and OCT features in spontaneously progressive idiopathic choriovitreal neovascularization. AB - A 47-year-old man presented with blurred vision in the right eye and was diagnosed as having idiopathic subfoveal choroidal neovascularization, which progressed to choriovitreal neovascularization in 2 weeks. Optical coherence tomography imaging revealed a subfoveal choroidal neovascularization breaking through the surface of an intact retina and growing as an epiretinal fibrovascular membrane accompanied by vitreomacular traction. Pars plana vitrectomy and removal of the choriovitreal neovascular membrane were performed. PMID- 17396698 TI - Retinal arterial macroaneurysm rupture following fundus fluorescein angiography and treatment with Nd:YAG laser membranectomy. AB - A 60-year-old woman with hypertension was seen for a routine examination. Her best-corrected visual acuity was 20/20 bilaterally. Intraretinal hemorrhages were found in the parafoveal region of the right eye. A fundus fluorescein angiography evaluation identified the hemorrhages as arterial macroaneurysms. The following day, the patient returned complaining of blurred vision that had developed the previous evening. Best-corrected visual acuity was counting fingers in the right eye. A subhyaloid hemorrhage approximately three optic disks in size was found in the macular region. The hemorrhage was drained with an Nd:YAG laser. PMID- 17396699 TI - Atypical myopic retinoschisis. AB - A 34-year-old woman with a refraction of-11 diopters in both eyes presented with a central scotoma and metamorphopsia in her right eye related to a juxtafoveal hemorrhage. Fluorescein angiography ruled out choroidal neovascularization and lacquer cracks. Optical coherence tomography revealed retinal splitting in the juxtafoveal area and the peripapillary area. No ophthalmoscopic peripheral abnormalities were observed. This is an atypical example of schisis associated with myopia because it was symptomatic, associated with hemorrhage, and did not affect the fovea. This may be an extension to the macula of a subclinical peripheral retinoschisis that became progressive and symptomatic due to vitreous tractional forces. PMID- 17396700 TI - Traumatic cataract surgery assisted by trypan blue. AB - The advantages of trypan blue use in anterior segment surgery have been well recognized, but few reports of its use in traumatic cataract surgery have been published. The authors describe three cases of traumatic cataract in which the role of trypan blue in performing safe and successful cataract surgery is demonstrated. Selective staining of the anterior lens capsule using trypan blue enabled the surgeon to identify the extent of structural damage and plan the surgery accordingly. In all three cases, the cataract was removed and the intraocular lens was implanted successfully without complications. Trypan blue is a safe and helpful aid in traumatic cataract surgery. PMID- 17396701 TI - Intravitreal bevacizumab for macular edema from idiopathic juxtafoveal retinal telangiectasis. AB - To assess the potential visual benefit of intravitreal bevacizumab in a patient with idiopathic juxtafoveal retinal telangiectasis refractory to focal laser treatment, an intravitreal injection of bevacizumab (1.25 mg) was given. Within 1 week, visual acuity improved from 20/50 to 20/25 and optical coherence tomography demonstrated complete resolution of macular edema. There was no adverse effect. The macular edema recurred after 3 months, requiring a repeat injection of bevacizumab with subsequent resolution of macular edema. An intravitreal injection of bevacizumab may provide potential short-term visual benefit in patients with macular edema from idiopathic juxtafoveal retinal telangiectasis. PMID- 17396702 TI - Identification of intraretinal neovascularization by high-speed indocyanine green angiography in idiopathic perifoveal telangiectasia. AB - Rapid advances in imaging technology have dramatically improved our understanding of the flow patterns of intraretinal circulation in normal and diseased states. To identify the angiographic features and flow pattern of retinal circulation in a patient with type 2 idiopathic perifoveal telangiectasia, dynamic simultaneous high-speed videoangiography using confocal scanning laser ophthalmoscopy was performed. This diagnostic tool provides enhanced anatomic resolution of retinal arterioles otherwise poorly defined by regular fluorescein and static indocyanine green angiography. High-speed indocyanine green angiography demonstrated dynamic flow abnormalities such as intraretinal neovascular complex and retino-retinal anastomosis in idiopathic perifoveal telangiectasia. PMID- 17396703 TI - Endoscopic cyclophotocoagulation using iris hooks versus viscoelastic devices. AB - Endoscopic cyclophotocoagulation utilizes a diode laser to ablate ciliary body epithelium, resulting in decreased intraocular pressure. Viscoelastic devices are often used to elevate the iris for improved exposure and efficient treatment of targeted tissue. Occasionally, early postoperative intraocular pressure spikes may occur due to retained viscoelastic material. Iris hooks may provide a safe alternative for elevation of the iris during endoscopic cyclophotocoagulation treatment and may be particularly advantageous in cases of aphakia or posterior capsule compromise in which viscoelastic removal is made more difficult. PMID- 17396704 TI - One-year outcomes of AquaLase cataract surgery. AB - The authors report surgical experience and clinical outcomes up to 1 year postoperatively in patients who underwent cataract surgery with the AquaLase liquefaction device (Alcon Laboratories, Fort Worth, TX). The device is a handpiece option for use with Alcon's Infiniti Vision System that uses heated balanced saline solution micropulses to liquefy lenticular material. Twenty-seven eyes of 23 patients underwent cataract extraction with the use of the AquaLase liquefaction device. The average age of participants was 68 years, and the average nuclear sclerotic grade was 1.96 on a 4-point scale. Outcomes were judged by metrics such as visual acuity, inflammation, endothelial cell count, and postoperative posterior capsule opacification. At 30 days postoperatively, 78% of eyes had a best-corrected visual acuity of 20/20. Visual acuity was 20/25 or better 1 year postoperatively in 88% of patients without complications except conversion to ultrasound phacoemulsification for two dense cataracts. PMID- 17396706 TI - Integration. PMID- 17396705 TI - Spontaneous visual improvement following a large traumatic submacular hemorrhage. PMID- 17396707 TI - Update on magnification and illumination in dentistry. PMID- 17396708 TI - A clinician's guide to purchasing surgical loupes. AB - Magnification using loupes is expanding within dental professions. Loupes offer enhanced vision of fine detail and critical ergonomic advantages. To achieve the maximum benefit from loupes, the dental professional should consider key features at the time of purchase. Proper fitting and adjustment are essential elements in the successful use of loupes. Though more expensive than simple, diopter lenses, the inherent advantages of compound and prism telescopic loupes make them superior choices when dental magnification is being considered. PMID- 17396709 TI - A review of the principles and practice of periodontal microsurgery. PMID- 17396710 TI - Magnification options in endodontics: a comparison of the operating microscope and the endoscope. PMID- 17396711 TI - Dental office waste disposal. Safety beyond the office. PMID- 17396712 TI - Oral and maxillofacial pathology case of the month. Melanocytic nevus, combined type (common blue). PMID- 17396713 TI - Mental health nursing is not fine art. PMID- 17396714 TI - Unusual & serious adverse effects of SSRIs: recognition & management. PMID- 17396715 TI - The transparency of aging. AB - This article is not meant to provide answers but to provoke thinking related to the questions we should be asking about the ethical personhood of aging adults. Are we covering over the rich opportunities to learn from their stories with an invisible cloak of transparency? Health care professionals have a moral obligation to rethink the assumptions that underlie their definitions of quality of life in aging. We cannot know what should be done unless we learn to listen to the life stories of aging people. This may even help us to see what is most real. PMID- 17396716 TI - Building a better mental health workforce: 8 core elements. AB - The field of behavioral health is facing a national crisis surrounding its workforce. Critical issues include problems in recruitment and retention and a serious lack of relevant preparation for work in mental health settings. This article identifies the challenges inherent in providing effective education and training to mental health staff who hold a bachelor's degree or less formal education. Key theories, concepts, and general principles of critical importance to all staff expected to work in a redesigned or transformed mental health system are described. Best and promising practices are contrasted with current practices, and specific recommendations including core concepts and competencies are listed, leading to the development of a training curricula targeted to meet these needs. PMID- 17396717 TI - Beyond the prescription pad: psychosocial treatments for individuals with schizophrenia. AB - Individuals with schizophrenia face multiple barriers to the achievement of optimal health, including apathy and poor concentration, sedative effects of medication, and poverty and lack of access to health education and treatment programs. In addition, the symptoms of the disease negatively affect their social functioning, which may result in isolation and lack of social support. In addition to providing psychiatric care and monitoring, both generalist nurses and advanced practice psychiatric nurses may assess physical and psychological risk factors, monitor medication responses, and provide health promotion to these individuals. Health promotion for individuals with schizophrenia includes psychosocial treatments to prevent relapse (e.g., education on the importance of treatment adherence) and to promote recovery (e.g., involving family members in treatment when possible). This article describes the key elements of recommended psychosocial treatments, briefly reviews the evidence for these treatments, and discusses clinical implications for all psychiatric nurses. PMID- 17396718 TI - Depression in school-aged children: assessment & early intervention. AB - A careful assessment of all of these factors will help nurse therapists who are also primary care providers develop a more holistic approach to treatment and, thus, increase treatment effectiveness. The three-pronged assessment approach discussed above is cost effective, safe, and indicated as a frontline measure to assess the risk for depression in children and their parents (particularly mothers), which can begin the process of "true" early intervention. I say "true" early intervention because in my role as an early intervention specialist, my first meeting with families usually occurs after a crucial incident requiring hospitalization of the child or the parent, or school failure, or both. My programs of intervention are hardly "early," although treatment is started relatively early in the life of the child. In the past few years, I have noticed that psychiatric-mental health advanced practice nurses, including myself, are increasingly working in some capacity in primary health centers, such as school based clinics, churches, collaborative practices, and community health centers. It would be wonderful if we used these settings to identify risk for depression in children and begin preventive treatment (starting with teaching good sleep hygiene, a treatment in itself) to truly make early intervention a reality. PMID- 17396719 TI - Anaphylaxis: my "top 10" list. PMID- 17396720 TI - The impact of wireless capsule endoscopy on gastrointestinal diseases. PMID- 17396721 TI - Gastric sarcoidosis. PMID- 17396722 TI - Ode to the hospitalist. PMID- 17396723 TI - Effects of noise contribute to physicians' challenges. PMID- 17396724 TI - How to improve the management of chest pain: hospitalists and use of prediction rules. AB - BACKGROUND: Three percent of patients with acute myocardial infarction are still missed despite the excess number of admissions with chest pain. The purpose of this study was to review the characteristics of patients who were admitted with chest pain, to evaluate the appropriateness of admissions and the outcomes. We also discuss whether use of a prediction rule could have made a difference in the management of these cases. METHODS: We performed retrospective chart review on all patients admitted to the hospitalist service with a diagnosis of chest pain. Each patient was risk stratified using Diamond and Forrester algorithm for probability of coronary artery disease (CAD), retrospectively. Results were analyzed using chi2 test or exact test and Student's t test. RESULTS: Of 260 patients admitted with chest pain to the hospitalist service, only 24 (9.2%) received the final diagnosis of acute coronary syndrome (ACS). The patients in the ACS group were older and more likely to be male and to have a history of hyperlipidemia, CAD, peripheral vascular disease, cerebrovascular disease and percutaneous coronary intervention (PCI). Of 34 patients who underwent cardiac catheterization, 20 (58.8%) had occlusive CAD and 14 of them received PCI. Risk stratification of patients, retrospectively, revealed 28.3% of the total patient population was high risk, while 6.6% of them were low risk. The number of ACS cases was highest in the high risk group, while none was detected in the low risk group. CONCLUSIONS: Our study demonstrated that using a prediction rule could have prevented about 6% of the chest pain admissions. Therefore, the use of risk stratification methods should be encouraged to decrease cost and improve efficiency of care. PMID- 17396725 TI - Trauma: life in the ER--education or entertainment? The patient's perspective. AB - OBJECTIVES: To determine whether patients felt that the show Trauma: Life in the ER has primarily educational or entertainment value. To determine if people felt that this show violated privacy. METHODS: Survey in an academic emergency department. Subjects were asked to participate in an initial survey, then asked to watch video clips from the show and complete a second survey. RESULTS: Three hundred ninety two participated in the initial survey. Fifty-six percent surveyed had seen the program, and of those, 55% stated that they watched the show for both educational and entertainment value. Of the initial participants, 267 watched the video and completed the second survey. Sixty-four percent stated that they would feel comfortable being filmed without prior permission. Eighteen percent of patients felt that the show violated patients' privacy. CONCLUSIONS: Most patients felt that this show was educational and did not invade their privacy. The majority were willing to be filmed for this television show, even without prior permission. PMID- 17396726 TI - Hyperglycemia and mortality in elderly patients with Staphylococcus aureus bacteremia. AB - OBJECTIVE: To investigate the association between hyperglycemia and in-hospital mortality in elderly patients with Staphylococcus aureus bacteremia (SAB). METHODS: We reviewed the medical records of 135 elderly patients with SAB admitted to two tertiary medical centers from January 2003 until December 2004. Patients were stratified into two groups: those with a 7-day post-SAB mean blood glucose < 170 mg/dL and those with a 7-day post-SAB mean blood glucose > or = 170 mg/dL. A stepwise logistic regression analysis was performed to determine whether the degree of hyperglycemia was a significant predictor of mortality. RESULTS: Seventy-four (54.8%) patients had methicillin-resistant Staphylococcus aureus bacteremia. During the follow-up period from admission until discharge, 36 (26.7%) patients died. Twenty-one (21.4%) of 98 patients with a 7-day post-SAB mean blood glucose < 170 mg/dL died, while 15 (40.5%) of 37 patients with a 7-day post-SAB mean blood glucose > or = 170 mg/dL expired. Multivariate analysis identified 3 independent determinants of death: Simplified Acute Physiology Score (SAPS) score at onset of SAB > 45 (OR 5.3, 95% CI {1.8, 15.5}, P = 0.002), a 7 day post-SAB mean blood glucose > or = 170 mg/dL (OR 3.3, 95% CI {1.2, 9.2}, P = 0.03), and altered mental status at the onset of SAB (OR 7.8, 95% CI {2.5, 23.9}, P = 0.0003). CONCLUSIONS: Hyperglycemia is an important marker of increased mortality among hospitalized elderly patients with SAB. PMID- 17396727 TI - Elevated red cell distribution width in the diagnosis of thrombotic thrombocytopenic purpura in patients presenting with anemia and thrombocytopenia. AB - OBJECTIVE: To determine if red cell distribution width (RDW) is elevated in thrombotic thrombocytopenic purpura (TTP) and to evaluate the sensitivity and specificity of such elevation in the diagnosis of TTP. METHODS: We retrospectively studied red cell distribution width at presentation in 25 consecutive patients with newly diagnosed TTP who were treated with plasmapheresis in our institution between 1997 to 2005. Control patients consisted of 20 consecutive patients without TTP who presented to the emergency room (ER) with anemia and thrombocytopenia. RESULTS: Patients with TTP had significantly elevated RDW compared with the controls (23.1% versus 17.2% respectively, P = 0.0001). An RDW > 18% had a 95% sensitivity and 70% specificity in the diagnosis of TTP, whereas RDW > 19% had 80% sensitivity and 85% specificity. CONCLUSIONS: TTP should be suspected in patients presenting with unexplained anemia and thrombocytopenia with an RDW > or = 18%. PMID- 17396728 TI - Group A Streptococcus: another resistant pathogen. AB - OBJECTIVES: In Greenville, South Carolina in 1992, erythromycin resistance in GAS was less than 5%, and there were no fully resistant strains. With a large increase in macrolide and azalide usage within the Greenville area, we again examined susceptibility patterns of pharyngeal GAS isolates in 2002 to 2003. METHODS: Community pediatric offices supplied 106 GAS isolates for study. Screening for macrolide resistance was done via Kirby-Bauer disk diffusion testing. Zones of inhibition from 16 to 20 mm were interpreted as intermediately resistant, and those 15 mm or less were interpreted as resistant per National Committee for Clinical Laboratory Standards guidelines. RESULTS: A total of 106 GAS isolates were tested; 0.9% of isolates were intermediately resistant to erythromycin and 11% were fully resistant. CONCLUSIONS: The rate of erythromycin resistance among GAS isolates has increased in the past 10 years in the Greenville community. This pattern has paralleled the increased utilization of macrolides in the same community. Continued monitoring of resistance rates will be needed to alert practitioners of possible treatment failures due to macrolide resistance. PMID- 17396729 TI - Early and late arrhythmogenic effects of doxorubicin. AB - BACKGROUND: To determine the incidence of early and late arrhythmogenic effects of doxorubicin-containing chemotherapy regimens. PATIENTS AND METHODS: A prospective study including 29 patients who were treated with doxorubicin containing regimens. Cardiac evaluation was based on 24-hour electrocardiographic monitorization (Holter), which was performed during the first cycle of doxorubicin-containing regimens, as well as after the last cycle of chemotherapy. RESULTS: The mean age of the patients was 45.8 +/- 15.1 (range 18-69). Holter records obtained during the first cycle of treatment revealed varying arrhythmias in 19 patients (65.5%) and in 18 (62.1%) patients after completion of therapy. One patient presented with syncope and both Mobitz Type 2 atrioventricular block and complete atrioventricular block were demonstrated. The patient subsequently underwent permanent pacemaker implantation. CONCLUSIONS: Doxorubicin may result in arrhythmias both in early and late periods of treatment. These arrhythmias are rarely life threatening. PMID- 17396730 TI - Reflux, dyspepsia, and disorders of the foregut. AB - Disorders of the foregut are an increasingly common cause of symptoms in Western populations. This review summarizes recent advances in the understanding and treatment of gastroesophageal reflux disease, dyspepsia and celiac disease. PMID- 17396731 TI - Small bowel capsule endoscopy: a systematic review. AB - Wireless capsule endoscopy offers a revolutionary diagnostic tool for small bowel diseases. Since its formal introduction, it has become an integral part of the diagnostic evaluation for obscure gastrointestinal bleeding. This relatively noninvasive imaging modality offered by small bowel capsule endoscopy is appealing to both patients and providers and consequently, the desire to expand its diagnostic role continues to grow. The use of CE in the diagnosis of Crohn disease and chronic diarrhea is being further investigated, as is the potential of employing this technique as a cancer surveillance mechanism in patients with hereditary polyposis syndromes which may involve the small bowel. This review article discusses the current indications for small bowel capsule endoscopy, the results of capsule endoscopy in patients with obscure gastrointestinal bleeding and small bowel diseases, and patient outcomes following capsule endoscopy. Capsule endoscopy is compared with traditional diagnostic modalities, including small bowel series, enteroclysis, CT, and push enteroscopy. Small bowel capsule endoscopy is the procedure of choice to evaluate obscure gastrointestinal bleeding, and is superior to radiographic procedures in detecting Crohn disease of the small bowel. PMID- 17396732 TI - Gastroparesis. AB - Gastroparesis is a symptomatic disorder of the stomach characterized by slow or delayed gastric emptying. Diabetes and idiopathic factors account for over 60% of gastroparesis cases. Symptoms associated with delayed gastric emptying include nausea, vomiting, abdominal bloating and early satiety. Delayed gastric emptying due to gastroparesis is managed by dietary adjustments, prokinetic medications, avoidance of medications that retard gastric motor activity and optimizing glycemic control in diabetic patients. Electrical stimulation and gastric pacing are an evolving treatment option for patients who do not respond to standard medical therapy. This article provides a review of gastric motility, the etiologies of gastroparesis and therapeutic approaches to this disorder. PMID- 17396733 TI - Noise pollution: a modem plague. AB - Noise is defined as unwanted sound. Environmental noise consists of all the unwanted sounds in our communities except that which originates in the workplace. Environmental noise pollution, a form of air pollution, is a threat to health and well-being. It is more severe and widespread than ever before, and it will continue to increase in magnitude and severity because of population growth, urbanization, and the associated growth in the use of increasingly powerful, varied, and highly mobile sources of noise. It will also continue to grow because of sustained growth in highway, rail, and air traffic, which remain major sources of environmental noise. The potential health effects of noise pollution are numerous, pervasive, persistent, and medically and socially significant. Noise produces direct and cumulative adverse effects that impair health and that degrade residential, social, working, and learning environments with corresponding real (economic) and intangible (well-being) losses. It interferes with sleep, concentration, communication, and recreation. The aim of enlightened governmental controls should be to protect citizens from the adverse effects of airborne pollution, including those produced by noise. People have the right to choose the nature of their acoustical environment; it should not be imposed by others. PMID- 17396734 TI - Anaphylactic shock: the great mimic. AB - Anaphylaxis, acute coronary syndrome and pulmonary embolism are conditions commonly seen in the acute medical setting which can be difficult to diagnose. Delay in establishing the correct diagnosis can result in either delayed or inappropriate treatment, and subsequent morbidity and mortality. The cases we present highlight the necessity of good clinical assessment when evaluating such patients. PMID- 17396735 TI - Four cases of patients with gastrointestinal granular cell tumors. AB - We present four cases of gastrointestinal granular cell tumors (GCT) with a literature review. Gastrointestinal granular cell tumors, a benign neural tumor thought to arise from Schwann cells, can occur in several areas, including the gastrointestinal tract. Studies suggest that endoscopic ultrasound and endoscopic removal is the treatment of choice for esophageal GCTs if they are small in size (< 2 cm) and do not involve the muscularis propria. GCTs are malignant less than 2% of the time. Although most GCTs are benign and can be followed endoscopically without resection, the malignant potential warrants evaluation with endoscopic ultrasound for possible endoscopic or surgical resection. PMID- 17396736 TI - Gastric sarcoidosis: a case report and review of the literature. AB - Sarcoidosis is a systemic granulomatous disease of unknown etiology that is characterized by the formation of noncaseating granulomas. Gastrointestinal (GI) tract involvement in sarcoidosis is rare. Gastric sarcoidosis, particularly involving the antrum, affects approximately 10% of patients with systemic disease. GI sarcoidosis commonly occurs subclinically, with clinical manifestations present in only 0.1 to 0.9% of patients with the disease. This is a rare case report of an individual with symptomatic gastric sarcoidosis. The patient presented with weight loss, nausea, and early satiety. An EGD and colonoscopy were performed and were grossly normal. However, biopsies of the gastric antrum revealed noncaseating granulomatous inflammation involving the gastric mucosa. Corticosteroid therapy was started and the symptoms abated almost immediately. We also offer a review of the literature. PMID- 17396737 TI - Ulceroglandular tularemia in a nonendemic area. AB - Two patients present with the abrupt onset of fever, malaise, anorexia, fatigue, progressive skin lesions and lymphadenitis. These patients represent two of the six cases of tularemia reported in Alabama over the last decade. The cases illustrate how mode of acquisition (direct versus vector-mediated) influences the clinical manifestations of ulceroglandular tularemia. In addition, a brief review of the epidemiology, differential diagnosis, clinical manifestations, and treatment of tularemia is provided. PMID- 17396738 TI - Massive multilocular cystic leiomyoma of the uterus: an extreme example of hydropic degeneration. AB - A 49-year-old woman underwent laparotomy for an undiagnosed, extremely large abdominal mass. At surgery, this was discovered to be a uterine tumor and she was subjected to a subtotal hysterectomy. Pathologic examination of the uterus revealed a multilocular cystic mass containing over 18 L of fluid that was established on histology to be a markedly hydropic leiomyoma. This case describes a very uncommon growth pattern of leiomyoma of the uterus that has only been occasionally described previously. We propose the name multilocular, cystic leiomyoma for this variant. PMID- 17396739 TI - McArdle disease presenting as acute renal failure. AB - In 1951, McArdle described a glycogen storage disorder which presents primarily as a myopathy. It is characterized by muscle pain, weakness and exercise intolerance with elevated creatine kinase from rhabdomyolysis. The pathophysiology involves a deficiency of myophosphorylase enzyme resulting in an inability to degrade glycogen stores. We present a novel case of McArdle disease (glycogen storage disease V [GSDV]) in a patient who had sickle cell trait and bulimia. The disease went unrecognized despite several admissions to the hospital with increased creatine kinase and muscle myopathy until the patient's initial presentation with acute renal failure. PMID- 17396740 TI - Emergency liver resection for combined biliary and vascular injury following laparoscopic cholecystectomy: case report and review of the literature. AB - A 75-year-old woman suffering from symptomatic cholelithiasis was admitted to our hospital for elective laparoscopic cholecystectomy (LC). Intraoperatively, because of severe inflammation and dense adhesions in the region of the Calot triangle and bleeding arising from the porta hepatis which obscured the operating field, the method was converted to a conventional open approach. Copious hemostasis was achieved using sutures, clips and diathermy, and no bile duct or vascular injuries were recognized intraoperatively. Because of severe right upper quadrant abdominal pain and significant deterioration of the liver function tests (LFTs) on the first postoperative day, the patient underwent a Doppler ultrasound scan which showed absence of blood flow at the level of porta hepatis. Urgent relaparotomy revealed an ischemic liver on the right, a transected common bile duct at the level of its confluence, a divided and ligated right hepatic artery and thrombosed portal vein down to its confluence. Thrombectomy and reconstruction of the portal vein were performed to salvage the left hemiliver, and after restoration of blood flow to the left hemiliver, a right hemihepatectomy and a Roux-en-Y hepaticojejunostomy on the left were performed. Liver resection serves an important role in the case of parenchymal necrosis due to combined biliary, hepatic artery and portal vein injury following laparoscopic cholecystectomy and moreover, the operation can be safely performed in the acute setting. PMID- 17396741 TI - Unrecognized delayed toxic lithium peak concentration in an acute poisoning with sustained release lithium product. AB - A 32-year-old female with a history of bipolar disorder was admitted after taking approximately 16 g of an extended-release lithium carbonate formulation in an attempted suicide. Five hours after consumption, the lithium serum level was 3.2 mEq/L. Fourteen hours after consumption, the lithium level was 5.1 mEq/L and the patient was asymptomatic. Due to a level > 4 mEq/L, the patient was transferred to a renal medicine service for hemodialysis. The lithium concentration 6 hours after the hemodialysis was 2.54 mEq/L. Thirty seven hours after the consumption (15 hours after hemodialysis), lithium levels increased up to 6.09 mEq/L. A second hemodialysis session was performed, which successfully reduced the serum lithium concentration to 1.86 mEq/L. Lithium levels 85 hours after the consumption were 0.61 mEq/L and the patient was transferred to the Psychiatry Department. Unrecognized delayed toxic peak lithium concentration may appear in an acute poisoning with a sustained release lithium product. Therefore, patients presenting with acute intoxication with extended release formulations should be managed with caution, and continued drug monitoring is suggested. PMID- 17396742 TI - Fatal passenger vehicle crashes 1999 to 2004 with drivers under age 15: the impact in Texas and other southern and southwestern states. AB - Texas has more fatal crashes involving unlicensed drivers under age 15 than does any other US state. Numbers and rates of such crashes are also above the national mean in many southern and Southwest states. Data on fatal passenger vehicle crashes from 1999 through 2004 were obtained from the online Fatality Analysis Reporting System (FARS). During the study period, there were 51 fatal passenger vehicle crashes in Texas in which drivers were under age 15. These crashes accounted for 12.3% of the US total. Nine southern states, including Texas, together accounted for 44% of all fatal crashes involving drivers under 15. Unlicensed crash rates per million inhabitants were higher in Texas than in other states with comparable populations but were much lower than those in other southern, southwest, and north central states. While Texas has recently improved its compliance with proposed graduated licensing models, state law explicitly prohibits police from stopping drivers based solely on age-related probable cause. This restriction may be a major barrier to effective detection and interdiction of under-age unlicensed driving. Because of the relatively high number of fatal crashes involving drivers under age 15 occurring in Texas, preventive efforts targeted to this state could modestly reduce the national burden of deaths due to very young unlicensed drivers. Expanding these efforts to other southern and southwest states could further reduce numbers and rates of such crashes. Expanded use of graduated licensing and increased public awareness are likely to prove effective tools in this public health effort. PMID- 17396743 TI - Yellow skin discoloration associated with sorafenib use for treatment of metastatic renal cell carcinoma. AB - This report describes a patient with metastatic kidney cancer who developed a deep yellow skin discoloration while on therapy with the oral multitargeted tyrosine kinase inhibitor (TKI), sorafenib. A significant hand-foot syndrome, featuring acral skin desquamation and tender erythema at pressure points, was also present. A thorough clinicolaboratory investigation did not reveal any evidence of jaundice, B12 deficiency, anemia, carotenemia, hypothyroidism, or any other disorder of endocrine or metabolic etiology. PMID- 17396744 TI - The role of positron emission tomography for the diagnosis and follow up of a patient with sarcoidosis and Takayasu arteritis. PMID- 17396745 TI - A rare cause of rhabdomyolysis. PMID- 17396746 TI - Bleeding Meckel diverticulum responds to intravenous pantoprazole. PMID- 17396747 TI - Glioblastoma multiforme disguised as sarcoidosis: pitfalls in diagnosis. PMID- 17396748 TI - Patient's page. Learning to manage heartburn and acid indigestion (reflux). PMID- 17396749 TI - Health care in Estonia. PMID- 17396750 TI - The challenge of migration and health. AB - Dr Manuel Carballo of the International Centre for Migration and Health provides the background to migration and its impact on the health of migrating peoples and the health care systems that treat them. PMID- 17396751 TI - Addressing the international, internal and public/private dimensions of the migration of health care workers: from awareness to action. PMID- 17396752 TI - Private sector initiatives to improve health in Africa. AB - In a presentation made to the Nigerian Chapter of IHF in Lagos in September 2006, Howard Lyons described an analytical approach developed with his colleagues in PA Consulting, Peter Classen and Shaun Bourgeois, to capture large volumes of internationally available data to reflect, at a macro-economic level, the attractiveness or otherwise of African countries for investment in health care. He then reported at a micro-economic level on initiatives and business propositions developed by African health care professionals living overseas who are seeking funding to implement their schemes in their native countries. He concludes that, as a way of improving health outcomes in Africa, private sector investment should be given more serious attention with particular reference to exploiting the talents and commitment of the African diaspora. PMID- 17396753 TI - Addressing the health workforce crisis: towards a common approach. AB - The challenges in the health workforce are well known and clearly documented. What is not so clearly understood is how to address these issues in a comprehensive and integrated manner that will lead to solutions. This editorial presents--and invites comments on--a technical framework intended to raise awareness among donors and multisector organizations outside ministries of health and to guide planning and strategy development at the country level. PMID- 17396754 TI - Use of appraisals for professional effectiveness. AB - Appraisal is usually linked with performance but it can also apply to intervention programmes. It is a distinct and formal management procedure used in the evaluation of work performance or the impact of a programme based on set objectives which are usually SMART. Every programme requires continual and meaningful feedback that transforms what looks technical into a dynamic and interactive process. This ties outcomes to intervention. Of great importance for a professional is self appraisal. When a professional is passion-driven to meet the needs of a client, the need to build capacity becomes obvious to remain in the market. Facility appraisal is as important as the environment of any service delivery and psychologically impacts on the consumer. Appraisal seems both inevitable and universal. To continually raise the standard of health care delivery, capacity building is at the crux. This will thoroughly equip health professionals for client-patient centered "best practices." PMID- 17396755 TI - Oral health in Uganda: the need for a change in focus. AB - This paper reviews the oral health care system of delivery in Uganda as accurately as possible given the limited research base. The paper looks into the evolution of dentistry in Uganda as well as the havoc wrecked on health care in general by two decades of civil strife in an effort to find explanations for the poor performance of the oral health system. Alternative methods of service delivery are suggested drawing from the rich experiences of countries like Sri Lanka and Tanzania. The alternative model highlights the need for a Primary Health Care approach to oral health service delivery implemented in a systematic manner. The paper concludes with a call to all Ugandan oral health workers to take personal and collective responsibility for the improvement of oral health outcomes of 24 million Ugandans through research, advocacy, adherence to professional ethics and continuing education. PMID- 17396756 TI - eHealth in Greece: strategic and implentation perpectives. AB - The main elements of the Greek Strategy for eHealth are presented, against a background overview of recent, relevant policy development. These elements, together with the current state of implementation were captured by the eHealth ERA survey on progress in individual Member States. An assessment of findings against targets set by the EU eHealth Action Plan and a reflection on the key success factors for converting policy into results are also presented. PMID- 17396757 TI - Developing health informatics as a recognised professional domain supporting clinical and health management activity. AB - This paper puts forward a case for use of the term "health informatics" to be deployed as a catalyst to collective recognition of the contribution that technology, information handling and decision support can provide to effective health care internationally. It cautions against disregarding the impact that the collective 'health informatics' may have on cohesion and recognition across the specialist clinical areas, management sectors and diverse professions involved in specialist areas. It also recognises that commonality of standards and consistency of protocols can be identified in many of the technical and clinical specialist areas, confirming the basis for a collective term. Having looked at potentially generic factors in the development of both a learned society and professional regulation body in the UK, the paper concludes that without recognition of the collective term "health informatics" there will be an grossy extended time frame before any of those working in this area gain the recognition and respect of a formal discipline. PMID- 17396758 TI - New online centre targets global nursing human resources challenges. PMID- 17396759 TI - Vocational training in Ireland. PMID- 17396760 TI - Commission consultation on health services. PMID- 17396761 TI - Basic dental care in Moldova. PMID- 17396762 TI - Local anaesthetic shortages continue to affect dentists. PMID- 17396763 TI - An audit of orthodontic referrals using IOTN as a comparison. AB - It is important that resources for orthodontic treatment within the Health Service Executive (HSE) are directed towards those children most in need of treatment. At present, children are referred using existing HSE guidelines. OBJECTIVES: To assess the level of treatment need in a sample of patients on the orthodontic waiting list in the North Eastern division of the HSE using the Index of Orthodontic Treatment Need (IOTN) as an objective comparison. Also, to compare these results with the findings of a similar audit in 2003 and to assess the effectiveness of recommendations from the 2003 audit. METHOD: Fifty models from each of two orthodontic units were selected. These were scored for the dental health component (DHC) and aesthetic component (AC) of IOTN by a calibrated examiner. RESULTS: In the 2005 audit, 100% of patients fell into DHC grades 4 or 5. These grades constitute a great need for treatment on dental health grounds. In the 2003 audit, 97% of patients fell into these two grades. The remaining 3% of children in 2003 were fostered and therefore entitled to orthodontic treatment under HSE guidelines. An average of 63.5% of patients fell into AC grades 8-10, i.e., deemed to be in great need of treatment on aesthetic grounds. CONCLUSION: The HSE screening guidelines identify patients in great need of orthodontic treatment using IOTN as an objective assessment of this need. The sensitivity of these guidelines requires assessment by measuring the level of unmet treatment need in 15-year-olds in the region. Recommendations arising from the 2003 audit relating to the filling of referral forms and the improvement of study model quality were found to have been effective. PMID- 17396764 TI - First permanent molars with molar incisor hypomineralisation. AB - Molar incisor hypomineralisation (MIH) is a common enamel defect presenting in the first permanent molars (FPM) and permanent incisors. This article presents the clinical findings and management considerations for the FPM with MIH to the general practitioner. The various treatment options are described with emphasis placed on early diagnosis as the most important prognostic factor. PMID- 17396765 TI - Single unit CAD/CAM restorations: a literature review. AB - Computer-aided design/computer-aided manufacture (CAD/CAM) has been used in dentistry since 1987. Since then, many CAD/CAM systems have been described, which enable the production of chair-side single unit dental restorations. These restorations are of comparable quality to those made by conventional techniques and have some specific advantages, including rapid production, improved wear properties, decreased laboratory fee and improved cross infection control. This literature review investigates the evidence base for the use of single unit CAD/CAM restorations. Materials, marginal gap, aesthetics, post-operative sensitivity, cementation, cost-effectiveness and longevity are discussed. PMID- 17396766 TI - Do you know your obligations? PMID- 17396767 TI - Seller beware. AB - Any dentist thinking of retiring needs to plan for the event, and that planning should include how to maximise Capital Gains Tax retirement relief. With assets having to be held for 10 years prior to retirement, that planning should start when the dentist reaches 45 years of age. As every dentist's circumstances are different, it is a tax that can become complicated. The best advice is to review your tax planning with your dental accountant as early as possible, and ensure you make the most of your practice assets. PMID- 17396768 TI - Convert profit into wealth. PMID- 17396769 TI - Chemical examination of Anacardium occidentale. AB - A new biflavonoid-C-glycoside named occidentoside, also the known ( )salipurposide and beta-sitosterol, have been isolated from the nut shells of Anacardium occidentale L. (Anacardiaceae). The structure of occidentoside has been established as tetrahydroninokiflavone-C-glucoside (III) from spectral and chemical evidence. This is the first biflavonoid to occur with one flavanone and one chalcone unit and also the first C-glycoside in the biflavonoid series. Identification of (-)salipurposide is also reported since it has been isolated for the first time from cashew nut shells. PMID- 17396770 TI - A new strain of Claviceps purpurea accumulating tetracyclic clavine alkaloids. AB - A new strain of Claviceps was isolated from a blokked mutant of Claviceps purpurea. This strain accumulates substantial amounts of clavine alkaloids (2 g/l). The alkaloid fraction is composed of chanoclavine-I ( approximately 10%) and a mixture of agroclavine/elymoclavine (90%). Most suitable for alkaloid production in submerged culture is an ammoncitrate/sucrose medium. The genealogy of the new strain, designated Pepty 695/ch-I is the following one: Pepty 695/S (ergotoxine producer) --> Pepty 695/ch (secoergoline producer) --> Pepty 695/ch-I (tetracyclic clavine producer). PMID- 17396771 TI - Studies in the field of drugs containing anthracene derivatives. AB - The anthranoid composition in roots of Rheum officinale has been followed during a one year cycle. No statistical correlation could be found between total anthranoid content and time of sampling. The mean total anthranoid content was found to be 3.17%, calculated on freeze dried plant material. On the other hand the ratio anthrone/total anthranoid varied with the season. It was concluded that all anthranoids were present under oxidized form, the anthraquinone, in summer months. They were in the reduced form, the anthrone, in winter. The conversion occured in a time lapse of three weeks. Also cyclic variations between the relative amounts of anthrone aglycones, anthronemono- and diglycosides could be observed. Just before the conversion anthrone --> anthraquinone and vice versa, the anthrone diglycoside content increased markedly. To find out if ambient temperature decrease is one of the factors influencing the anthraquinone --> anthrone conversion, analyses were carried out on plants submitted to a cooling treatment. After comparison with reference plants, it was clear that the composition was similar to that of plants harvested in winter time. This experiment confirms that the conversion anthraquinone --> anthrone could be induced artificially by decreasing ambient temperature. PMID- 17396772 TI - Constituents of local plants. AB - The antispasmodic principle of Cymbopogon proximus has been identified as cryptomeridiol (5) from its spectral properties and comparison with an authentic sample. PMID- 17396773 TI - Anti-tumour effects of the sesquiterpene Lactone parthenin. AB - The anti-tumour effects of the sesquiterpene lactone, parthenin was studied both in vivo and in vitro. Parthenin showed marked cytotoxic activity to P815 mastocytoma, L1210 leukemia, and M-1 rhabdomyosarcoma cells in vitro. In vivo studies showed intraperitoneal drug administration could either cure mice or increase their survival time after intraperitoneal of subcutaneous injection with tumour cells. PMID- 17396774 TI - Vallesiachotamine and isovallesiachotamine from the seeds of Strychnos tricalysioides. AB - The alkaloids vallesiachotamine and isovallesiachotamine have been isolated from the seeds of Strychnos tricalysioides and identified on the basis of their spectral characteristics. The potential chemotaxonomic significance of these alkaloids is briefly discussed. PMID- 17396775 TI - Brine shrimp: a convenient general bioassay for active plant constituents. AB - A method, utilizing brine shrimp (Artemia salina Leach), is proposed as a simple bioassay for natural product research. The procedure determines LC (50) values in microg/ml of active compounds and extracts in the brine medium. Activities of a broad range of known active compounds are manifested as toxicity to the shrimp. Screening results with seed extracts of 41 species of Euphorbiaceae were compared with 9KB and 9PS cytotoxicities. The method is rapid, reliable, inexpensive, and convenient as an in-house general bioassay tool. PMID- 17396776 TI - [Papaver bracteatum, influence of cold treatment and cloning on Thebaine content of the capsules.]. AB - A high heterogeneity of thebaine yields is revealed in capsules of plants obtained from seeds cultivated in a phytotron. Clonage methods could be used to perform culture. Heterogeneity of plants hides influence of low temperature on thebaine production. With clone this influence appears to be slight but appreciable. PMID- 17396777 TI - [Alkaloid Content of Thalictrum minus from the Slavjanka Mountains.]. AB - In Thalictrum minus plants, collected in the Slavjanka Mountains, the alkaloids thalmelatine, thalipine, ocoteine, berberine, thalicarpine and glaucine were quantitatively determined by HPLC. PMID- 17396779 TI - A New isoflavanone phytoalexin from Medicago rugosa. AB - An isoflavonoid phytoalexin produced by the fungus-inoculated leaflets of Medicago rugosa Desr. has been formulated as (+/-)-7,4'-dimethoxy-2' hydroxyisoflavanone (isosativanone). This compound cooccurs with the known isoflavonoid fungitoxins, (-)-medicarpin, (-)-vestitol, (-)-isosativanand(+/-) vestitone. PMID- 17396778 TI - Cyanogenesis in Passifloraceae. AB - A novel cyclopentenoid cyanogenic bis-glycoside has been isolated from Passiflora capsularis L. Its structure has been tentatively determined as 4-boivinosyl tetraphyllin B. The name passicapsin is proposed for this compound. Linamarin has been isolated from Passiflora warmingii Mast. Extracts of Passiflora perfoliata L. yielded a (probably glycosidic) cyclopentenone dervative, which possibly originated from an unstable glycoside having a free alpha-hydroxynitrile group. PMID- 17396780 TI - Anthraquinones of Cassia marginata seeds. AB - From the defatted seeds of Cassia marginata fam (Caesalpinaceae) two new anthraquinone glycosides have been isolated and characterised as physcion-8-O alpha-L-xylopyranoside and emodin-8-O-alpha-L-arabinopyranoside. PMID- 17396781 TI - Aristolochic acids in Aristolochia chilensis. AB - The major components of the acid fraction of the leaves and tender stems of Aristolochia chilensis Miers are aristolochic acids I and Ia. Aristolochic acid Ia has been isolated from plant material for the first time, and its PMR spectrum is discussed. PMID- 17396782 TI - Thebaine from root cultures of Papaver bracteatum. AB - Root cultures derived from cell suspension cultures of Papaver bracteatum were shown to produce thebaine (yield 0.03%). PMID- 17396783 TI - [Presence de Davanone et de Deux Autres Sesquiterpenes a Noyau Furane dans l'Huile essentielle d'Artemisia abrotanum L]. PMID- 17396784 TI - Initiation, maintenance and alkaloid content of Catharanthus roseus leaf organ cultures. PMID- 17396785 TI - Constituents of Magnolia grandiflora i: mono-O-methylhonokiol. PMID- 17396786 TI - [Neue Flavonglykoside in den Blattern von Digitalis lanata]. PMID- 17396787 TI - Chemical constituents of Swertia paniculata. PMID- 17396788 TI - Coleonol and forskolin from Coleus forskohlii. PMID- 17396791 TI - [New cardenolides of Convallaria majalis.]. AB - A detailed study of the cardiac glycosides of Convallaria majalis L. allowed the detection and structure determination of ten new cardenolides. Four glycosides of bipindogenin (bipindogenin-3-0-6'-deoxy-beta-D-guloside, bipindogenin-3-0-6' deoxy-beta-D-alloside, bipindogenin-3-0-6'-deoxy-beta-D-allosido-alpha-L rhamnoside and bipindogenin-3-0-beta-D-alloside) were isolated as well as three derivatives of sarmentologenin (sarmentologenin-3-0-alpha-L-rhamnoside = sarhamnoloside, sarmentologenin-3-0-6' -deoxy-beta-D-guloside and sarmentologenm 3-0-6'-deoxy-beta-D-alloside) and three isomeric glycosides of sarmentosigenin A (sarmentosigenin-3-0-alpha-L-rhamnoside = tholloside, sarmentosigenin-3-0-6' deoxy-beta-D-guloside and sarmentosigenin-3-0-6'-deoxy-beta-D-alloside). The occurrence of allose, and of the disaccharide rhamnosido-6-deoxy-allose, resp. as the sugar moieties of cardiac glycosides, was detected for the first time. PMID- 17396795 TI - Stem bark alkaloids of Rauwolfia vomitoria. AB - Forty-three indole alkaloids were isolated from 7 kg Rauwolfia vomitoria stem bark; 39 were identified and 2 partially characterised. At least 72 alkaloids classified into 19 types occur in the plant and the interrelationship of the alkaloids and their distribution throughout the plant has been discussed. The major alkaloids of the stems were heteroyohimbines (especially reserpiline) and N (a)-demethyldihydroindoles. PMID- 17396796 TI - Przewalskia tangutica - A tropane alkaloid-containing plant. AB - Przewalskia tangutica Maxim. (Solanaceae), a plant of Tibetan traditional medicine, was investigated. Contents of tropine, cuscohygrine, (-)-6beta hydroxyhyoscyamine, hyoscyamine, apoatropine, scopolamine and daturamine were determined (Table I). The roots of this plant showed high contents of both hyoscyamine (major constituent) and total alkaloids, amounting to 1.67-3.82% and 2.06-4.01%, respectively. This plant and the crude drugs prepared from it are noteworthy as new sources of tropane alkaloids. The occurrence of tropane alkaloids in solanaceous plants of China is discussed briefly (Table II). PMID- 17396797 TI - Studies on Sudanese medicinal plants III. AB - A prelimenary study was carried out to evaluate the potential of cultivating Hyoscyamus muticus L. as a source of hyoscyamine. The alkaloid yield was found to be quite favorable for the commercial utilization of this plant. PMID- 17396798 TI - Alkaloids from Fumaria parviflora and F. kralikii. AB - From Fumaria parviflora Lam. were isolated the major alkaloids protopine and adlumidiceine and the minor alkaloids parfumine, fumariline, dihydrofumariline, cryptopine, (-)-stylopine, 8-oxocoptisine, sanguinarine, and oxysanguinarine. The quaternary protoberberine fraction gave coptisine. Adlumidiceine, dihydrofumariline, 8-oxocoptisine, sanguinarine, and oxysanguinarine were found in F. parviflora for the first time. F. kralikii Jordan gave, besides the major alkaloids protopine, fumarophycine, and O-methylfumarophycine, the alkaloids adlumidiceine, berberine, coptisine, cryptopine, (-)-stylopine, and (-)-canadine. Only protopine and cryptopine have been isolated from F. kralikii earlier. PMID- 17396800 TI - [Barbatoside, a new Iridoid glucoside from Penstemon barbatus]. AB - From leaves of Penstemon barbatus Nutt. a new iridoid glucoside has been isolated. The structure was established mainly by spectroscopic methods ( (1)H NMR, (13)C-NMR, IR, MS). PMID- 17396801 TI - [Helenalin und seine Derivate, die herzwirksamen Verbindungen der Arnikabluten]. PMID- 17396802 TI - Coronary vasodilatory, spasmolytic and cAMP-Phosphodiesterase inhibitory properties of dihydropyranocoumarins and dihydrofuranocoumarins. PMID- 17396803 TI - [Zur Standardisierung von Phytopharmaka: Nachweis von herzaktiven Verbindungen in Phytopharmaka]. PMID- 17396804 TI - [Testsystem zur Bestimmung der in vitro-Wirksamkeit neuer Herzglykoside]. PMID- 17396805 TI - [Weitere Sesquiterpenlactone aus den Bluten von Arnica chamissonis]. PMID- 17396806 TI - [Uber die sedative Wirkung des Weissdorns]. PMID- 17396807 TI - [Zum Mechanismus der positiv inotropen und gefasserweiternden Wirkung von Forskolin aus Coleus forskohlii]. PMID- 17396808 TI - Principles regulating cardenolide formation in cell cultures of digitalis lanata. PMID- 17396809 TI - [Der Einfluss des Alterns auf die Sekundarstoffbildung in Gewebe-Kulturen aus Digitalis purpurea]. PMID- 17396810 TI - Effect of nutrients deficiency on organic constituents in digitalis lanata. PMID- 17396811 TI - On the pharmacology of the alkaloid tetramethoxyaporphine and of its semisynthetic structural analogs. PMID- 17396812 TI - [Veranderungen der Steroidgehalte in Digitalis lanata unter Wasserstress]. PMID- 17396813 TI - [Zur Kenntnis der Peroxidase-Aktivitat und der Peroxidase-Isoenzymmuster von Gewebekulturen aus Digitalis purpurea und Digitalis lanata]. PMID- 17396814 TI - Effect of environment on the growth and glycosides of callus tissue of digitalis lanata. PMID- 17396815 TI - [Zwei neue lebertoxische Pyrrolizidinalkaloide aus Gynura scandens]. PMID- 17396816 TI - [Vergleich der genotoxischen Aktivitat von Aristolochiasaure in verschiedenen Testsystemen]. PMID- 17396817 TI - The neuromuscular blocking actions of coclaurine derivatives and of paeoniflorin derivatives. PMID- 17396818 TI - [Wirkung von rizinolsaure auf die prostaglandin-biosynthese in blutgefassen]. PMID- 17396819 TI - [Zum Mechanismus der laxierenden Wirkung von Oleum Ricini]. PMID- 17396820 TI - Phytochemical and molluscicidal studies on Hedychium gardnerianum and its possible use in control of Bilharzia. PMID- 17396821 TI - Isolation of a new flavone and some essential oils with antimicrobial and antifungal properties from the stem bark of Jatropha podagrica. PMID- 17396822 TI - Antistress plant drugs. A new approach in the treatment of stress diseases. PMID- 17396823 TI - Protective effect of G2 102 on life supporting a study of white mouse under neutron irradiation. PMID- 17396824 TI - [Beitrag zur chemischen Zusammensetzung des dalmatinischen Salbeiols]. PMID- 17396825 TI - [Validierung in der arzneipflanzenanalytik]. PMID- 17396826 TI - Gentamycin und Lincomycin induzierte mitochondriale Strukturanomalien an menschlichen embryonalen Lungenfibroblasten in vitro. PMID- 17396827 TI - Phytochemical investigation of Melilotus siculus and M. indica. PMID- 17396828 TI - [Analyse der Hydrazone von Gyromitra esculenta, mittels HPLC, Differentialpuls und AC-Polarographie]. PMID- 17396829 TI - [Bestimmung des Galegins und die antidiabetische Wirkung der Droge Herba Galegae]. PMID- 17396830 TI - Effect of surface active agent on zeta potential of cells in callus cultures of Trigonella Foenum-graecum. PMID- 17396831 TI - Conversion of elymoclavine to paspalic acid by a particulate fraction from an ergotamine-producing strain of Claviceps sp. PMID- 17396832 TI - Aberrant biosynthesis of opium alkaloids: the effects of structural modifications of reticuline on its racemization and biotransformation. PMID- 17396833 TI - [Ungesattigte Saureamide - ein chemisches Merkmal der Gattung Achillea]. PMID- 17396834 TI - [Neue iridoidglykoside der gattung penstemon]. PMID- 17396835 TI - [Neue cyanglykoside aus compositen]. PMID- 17396836 TI - [Der Einsatz von Prometryn und Desmetryn bei Thymus vulgaris, Salvia officinalis und Silybum marianum und ihre Auswirkung auf Ertrag und Inhaltsstoffe]. PMID- 17396837 TI - 3,3'-Di-O-methyl ellagic acid-4-O-glucuronosyl-arabinosyl-arabinosyl-glucoside from Prosopis juliflora pods. PMID- 17396838 TI - Isolation procedure and structure of new saponins from Fagonia indica. PMID- 17396839 TI - Revised structure of phaeantharine. PMID- 17396840 TI - [Tagesrhythmische schwankungen im alkaloidgehalt von pflanzen]. PMID- 17396841 TI - [Untersuchungen uber Triacin-Ruckstande in Thymus vulgaris und Salvia officinalis nach Anwendung der Herbicide Gesagard 50 und Semeron 25.]. PMID- 17396842 TI - [Charakterisierung eines neuen 4-Arylcumarinderivates aus Copalchirinde]. PMID- 17396843 TI - Alkaloids of Senecio desfontainei (= S. coronopifolius). PMID- 17396844 TI - Compensation pattern of essential oil yield components in the selected caraway genotype 1. 4. 36. PMID- 17396845 TI - Anthraquinones of Hemerocallis aurantiaca. PMID- 17396846 TI - [Zur antibakteriellen Wirkung von Clavinalkaloiden und deren partialsynthetischen Derivaten]. PMID- 17396847 TI - Flavonoids of Francoeuria crispa. PMID- 17396848 TI - [Pharmakokinetik von 7,3', 4'-Tri-O-(2-hydroxyethyl)-rutin (Troxerutin) nach oraler Applikation am Menschen]. PMID- 17396849 TI - Biflavonoids and glycosides of Garcinia kola stem bark. PMID- 17396850 TI - [Chemische Analyse der Wurzelextrakte von Rhaponticum carthamoides]. PMID- 17396851 TI - New Alkaloids from Haplophyllum glabrinum and Ruta graveolens. PMID- 17396852 TI - [Chamaecytisus-Drogen als Rohstoff fur Spartein]. PMID- 17396853 TI - [Entgiftung von Absinthkraut durch CO2-Hochdruckextraktion]. PMID- 17396854 TI - [Ecdysteroid-fuhrende einheimische Chenopodiazeen und Caryophyllazeen]. PMID- 17396855 TI - Aliphatic carbonyl-derivatives (attractants) in the flowers and fruits of Evodia hupehensis. PMID- 17396856 TI - Microscopical study of valerian plants in vivo and in vitro. Histochemical localization of valepotriates. PMID- 17396857 TI - Cytotoxic anhydronium bases from Strychnos usambarensis. PMID- 17396858 TI - Anti-atherogenic action of panax ginseng in rats and in patients with hyperlipidemia. PMID- 17396859 TI - [Untersuchungen zur Analytik und Biosynthese der Glycyrrhizinsaure in Glycyrrhiza glabra]. PMID- 17396860 TI - [Der Einfluss von Valerenal und Valerensauren aus Valeriana officinalis auf die Aktivitat von Ratten und den Dopamin-Metabolismus im Rattengehirn]. PMID- 17396861 TI - Analysis of primary and secondary plant products in Valeriana officinalis callus during a growth cycle. PMID- 17396862 TI - [Trennung der diastereomeren Aloine mittels Droplet-Counter-Current Chromatography]. PMID- 17396863 TI - Two new secoiridoid glucosides from Syringa vulgaris. PMID- 17396864 TI - [Uber Inhaltsstoffe aus Uvaria scabrida]. PMID- 17396865 TI - [Ein Beitrag zur Pharmakologie und Toxikologie einer Drogenkombination und ihrer Einzeldrogen]. PMID- 17396866 TI - A standardized HPLC method for analyses of lichen compounds from the genus cladonia. PMID- 17396867 TI - [Cardenolide aus Ornithogalum boucheanum und Ornithogalum balansae]. PMID- 17396868 TI - New iridoids from Tecoma heptaphylla. PMID- 17396869 TI - 6-O-beta-D-xylopyranosylcatalpol, a new iridoid glycoside from Verbascum thapsiforme. PMID- 17396870 TI - Continuous biotransformation by immobilized cells in bioreactors. PMID- 17396871 TI - Cryopreservation of Digitalis lanata cells cultivated in vitro. PMID- 17396872 TI - Accumulation of purpureaglycoside a in vacuoles of Digitalis lanata cells cultivated in vitro. PMID- 17396873 TI - [Rutacridonepoxide in Suspensionskulturen von Ruta graveolens. Einfluss von Elicitoren und Mischkultur mit Hefen]. PMID- 17396874 TI - Indole alkaloids from cell suspension cultures of Stemmadenia tomentosa and Voacanga africana. PMID- 17396875 TI - Clonal multiplication of Digitalis lanata by meristeme culture. PMID- 17396876 TI - [Zur Kenntnis des Cardenolid- und Flavonoidmusters der Blatter von Digitalis purpurea wahrend verschiedener Entwicklungsstadien]. PMID- 17396877 TI - [In vitro-Kulturen von Arctostaphylos-Arten 2. Variabilitat des Arbutin- und des Methylarbutingehaltes bei Arctostaphylos uva ursi und Arctostaphylos nevadensis]. PMID- 17396878 TI - Teucrioside, a new phenylpropanoid glycoside from Teucrium chamaedrys. PMID- 17396879 TI - Structure and biological activity of sesquiterpene lactones from Thapsia species and of analogue derivatives. PMID- 17396881 TI - Presence of phthalides in umbelliferous plants. PMID- 17396880 TI - Substituent induced chemical shifts for aromatic carbons in indole alkaloids. PMID- 17396882 TI - Variation in the asperuloside production of Galium verum. PMID- 17396883 TI - Acetophenone glycosides from Veronica species. PMID- 17396884 TI - Structure analysis of eukovoside, a new phenylpropanoid glycoside from Euphrasia rostkoviana. PMID- 17396885 TI - [Cumarinderivate aus den Wurzeln von Eryngium campestre]. PMID- 17396886 TI - Literature survey of the genus Tabernaemontana. PMID- 17396887 TI - [Untersuchungen zur Verstoffwechslung von Benzylisothiocyanat beim Menschen]. PMID- 17396888 TI - Gentiogenal, a new antimicrobial iridoid derived from gentiopicrin (gentiopicroside). PMID- 17396889 TI - Investigation of commercial plant extracts for photoactive furocoumarins. PMID- 17396891 TI - [Die Flavonoidmuster in den Organen von Digitalis lanata]. PMID- 17396890 TI - Radioimmunoassay of lysergic acid derivatives from Claviceps paspali fermentation cultures. PMID- 17396892 TI - Application of NMR techniques in the quantitative analysis of tropane alkaloids and sennosides. PMID- 17396893 TI - Pharmacological active alkaloids of Pterotaberna inconspicua. PMID- 17396894 TI - Phenolics and other constituents from Eclipta alba. PMID- 17396895 TI - [Hagebutten: Unterscheidungsmoglichkeit durch Dunnschichtchromatographie]. PMID- 17396896 TI - Flavonoids from fruits of Silybum marianum var. albiflora. PMID- 17396897 TI - Quantitative determination of beta-methyldigitoxin and beta-methyldigoxin by HPLC in fermenter broth. PMID- 17396898 TI - Effects of Melia azedarach extract against schistosomiasis in mice. PMID- 17396899 TI - [Neue Chromenoflavanone aus der Wurzelrinde von Amorpha fruticosa]. PMID- 17396900 TI - Isolation and identification of alkaloids from Arundo donax. PMID- 17396901 TI - Pharmacological survey of Nigerian medicinal plants used as anticonvulsants for cardiovascular activity. PMID- 17396903 TI - [Histochemische Methode zum Nachweis der Lokalisation von Fenoloiden (Flavonoiden Rotenoiden)]. PMID- 17396902 TI - Effects of hypoxanthine-9-L-arabinofuranoside on isolated coronary artery of the goat. PMID- 17396904 TI - Formation of secondary products derived from tyrosine in Papaver suspension cultures. PMID- 17396905 TI - A contribution to the molluscicidal activity of Hedychium gardniarum. PMID- 17396906 TI - [Two new Dammaran sapogenins from leaves of Panax notoginseng.]. AB - Hydrolysis of the crude saponin extracted from the leaves of Panax notoginseng (Burk.) F. H. Chen yields five sapogenins which are separated by column chromatography. Two of these are identified to be panaxadiol and panaxatriol whose structures were elucidated earlier. The third sapogenin ist found to be Dammar-20(22)en-3beta, 12beta,-26-triol by carbon-13 NMR. The structure of the fourth sapogenin is established by X-ray diffractometry and carbon-13 NMR to be 20 ( R)-Dammaran-3beta, 12beta, 20, 25-tetrol. PMID- 17396911 TI - [New cardenolides from Convallaria majalis]. AB - From the leaves of Convallaria majalis L. eight glycosides of strophanthidin, cannogenol, sarmentogenin and 19-hydroxysarmentogenin, resp., were isolated and structurally elucidated as strophanthidin-3-O-6'-deoxy-beta-D-allosido-alpha-L rhamnoside, strophanthidin-3-O-6'-deoxy-beta-D-allosido-alpha-L-arabinoside, strophanthidin-3-O-alpha-L-rhamnosido-2'-beta-D-glucoside, cannogenol-3-O-6' deoxy-beta-D-allosido-beta-D-glucoside, cannogenol-3-O-6'-deoxy-beta-D-allosido alpha-L-rhamnoside, 19-hydroxy-sarmentogenin-3-O-alpha-L-rhamnoside, sarmentogenin-3-O-6'-deoxy-beta-D-allosido-alpha-L-rhamnoside and sarmentogenin-3 O-6'-deoxy-beta-D-guloside, mainly by means of their H-NMR-spectra, by EI-MS and LD-MS. This is the first report of the occurrence of 19-hydroxy-sarmentogenin in nature. The presence of arabinosido-6-deoxyallose within the sugar chain of a cardiac glycoside was detected for the first time, as well. Including the new glycosides, a total of 38 cardenolides, derived from nine aglycones, have now been found in Convallaria majalis. PMID- 17396912 TI - [Cardiac glycoside composition in the leaves of Digitalis schischkinii]. AB - Cardenolide glycosides of stem leaves of D. schischkinii, at the same age and location, were analysed quantitatively over several vegetation periods and the limit values found are tabulated. Total glycoside content of the leaves was 0.86 1.0%. Digoxigenin glycosides was 50% and the sum of lanatosid C, desacetyllanatosid C, acetyldigoxin and digoxin was 40% of the content. As yet unknown glycosides diginatigenin-digilanidobioside, the corresponding secondary glycoside and diginatigenin-bisdigitoxoside were identified as minor compounds. PMID- 17396914 TI - [Qualitative and Quantitative HPLC-Analysis of Cardiac Glycosides]. AB - An analytical pathway for identification of cardenolides from Adonis, Convallaria, Strophanthus and Lophopetalum extracts by HPLC is described. Structure-retention-relationships between cardiac glycosides and HPLC-separations of Strophanthus gratus, Strophanthus kombe and Adonis vernalis drugs are discussed. PMID- 17396916 TI - [The sedativ-hypnotic principle of hops]. AB - A gas chromatographic method for the quantitative evaluation of 2-methyl-3-butene 2-ol, a sedative hop constituent, is described. Depending on the storing time, the content of this compound was determined in a large number of hops, hop extracts, and drugs. While only traces of the compounds were detected in fresh hops, the concentration continously increased after drying to reach maximum levels (approx. 0.15%) within 2 years of storing at room temperature, irrespective of the specific kind. Hop extracts standardized to alpha-acids, which are available on the market, were found to contain relatively small amounts of 2-methyl-3-butene-2-ol. Only traces of the compounds were discovered in hop containing "sedative dragees", whereas a relatively high concentration was found in a sedative bath preparation. Thus, pharmacologically relevant concentrations of 2-methyl-3-butene-2-ol can be reached in both tea and bath preparations. However, since there is a reason to suppose that 2-methyl-3-butene-2-ol may also be formed IN VIVO by metabolization of alpha-acids, negative conclusions as to the effectiveness of the dragees should not be drawn before completion of the current studies. PMID- 17396919 TI - Application of radioimmunoassay to the quantitation of hyoscyamine in plant material. AB - A radioimmunoassay has been developed for the analysis of atropine ( DL hyoscyamine) from unpurified ethanolic extracts of Atropa belladonna L. The antiserum used was highly specific for D-hyoscyamine, the unnatural enantiomer. Consequently, a racemization step had to be included in the assay procedure in order to obtain the analysis of atropine. The method is rapid and sensitive, giving excellent values for accuracy, precision and correlation. There was practically no interference in the assay from other compounds present in the extracts. PMID- 17396920 TI - Detection of some alkaloids on thin layer chromatograms using pi-acceptors. AB - The reactions of chloranil, fluoranil, 2,5 dichlorobenzoquinone and chloranilic acid with some alkaloids on thin layer chromatograms were examined with a view to determine their suitability as visualizing agents. Chloranil and 2,5 dichlorobenzoquinone (each 0.5% w/v in dioxan) were found to be most suitable as they gave coloured reaction products with many alkaloids. PMID- 17396921 TI - Flavonoids and rotenoids from Tephrosieae and related tribes of leguminosae. AB - Seeds and/or roots of eleven species of Leguminosae, mainly Tephrosiae, were examined. Prenylated chalcones and flavanones were isolated from Lonchocarpus nitidus and L. neuroscapha seeds; rotenoids from Lonchocarpus spruceanus, Piscidia mollis and a Tephrosia sp.; demethylhomopterocarpin from Canavalia bonariensis (Phaseoleae). No flavonoids or rotenoids were found in L. nelsii, L. rugosus, T. multijuga (seeds), Millettia grandis (Tephrosieae) and in Xeroderris stuhlmanii (Dalbergieae). PMID- 17396922 TI - Deoxyandrographolide-19beta-D-glucoside from the leaves of Andrographis paniculata. AB - A new diterpene glucoside has been isolated from the leaves of Andrographis paniculata and its structure elucidated as deoxyandrographolide-19beta-D glucoside on the basis of chemical and spectral evidence. A detailed proton magnetic resonance study of the diterpene was crucial in the structure determination. PMID- 17396923 TI - Flavonoids of Thelesperma megapotanicum. PMID- 17396924 TI - Biogenesis of the ajmaline type alkaloids. PMID- 17396925 TI - Carbon-13 spectral data of prenylated coumarins. PMID- 17396928 TI - [Sur la Preparation Aisee d'Hydroxy-17 Dihydro-16,17 Heteroyohimbines]. PMID- 17396927 TI - Chemical investigation of Cocculus leaebe. PMID- 17396930 TI - Immobilization of strictosidine synthase from Catharanthus cell cultures and preparative synthesis of strictosidine. AB - Strictosidine synthase was partially purified from Catharanthus roseus cell suspension cultures and immobilized on CNBr-activated Sepharose. The immobilized enzyme exhibits a thermostability increased 300 fold over that of the soluble enzyme and catalyses exclusively the formation of the 3alpha(S)-isomer, strictosidine. Gram quantities of this biologically active glucoalkaloid, which hitherto had been difficult to synthesize and purify, were prepared. PMID- 17396931 TI - The effects of plant growth regulators and culture conditions on the growth and the alkaloid content of callus cultures of Cinchona pubescens. AB - Studies were performed on the growth and the alkaloid content of stem callus cultures of Cinchona pubescens Vahl concerning the influence of plant growth regulators in the medium. After two months the increase of fresh weight of cultures grown in the dark was less than those incubated with illumination. A number of quinoline and indole alkaloids could be detected by means of TLC. Media F (zeatin: 1 microM, indole-3-butyric acid: 1 microM) and H (zeatin: 1 microM, 2,4-dichlorophenoxy acetic acid: 1 microM) showed the best alkaloid production and tissue growth. PMID- 17396932 TI - Identification of the alkaloids and anthraquinones in Cinchona ledgeriana callus cultures. AB - The isolation of quinoline alkaloids, indole alkaloids and anthraquinones present in callus cultures initiated from different parts of Cinchona ledgeriana plants is described. From leaves of C. ledgeriana a new minor alkaloid was isolated and identified as 10-methoxycinchonamine. PMID- 17396933 TI - Alkaloid production by leaf organ, root organ and cell suspension cultures of Cinchona ledgeriana. AB - The alkaloids produced by leaf organ, root organ and root suspensions of two varieties of Cinchona ledgeriana have been determined quantitatively by HPLC. In contrast to previous findings, the root organs and root suspensions yielded quinine and quinidine. The root cells release alkaloids into the suspension medium. PMID- 17396934 TI - Constituents of the cultivated mulberry tree. AB - In addition to mulberrofuran C, a new 2-arylbenzofuran derivative, six flavonoid derivatives, cyclomorusin, morusin, kuwanon C, E, G and H, as well as a known 2 arylbenzofuran derivative, chalcomoracin, were isolated from extracts of root bark of the cultivated mulberry tree (a variety of Morus bombycis K OIDZUMI). The structure of mulberrofuran C was shown to be VIII on the basis of spectral data. It is regarded biogenetically as a Diels-Alder adduct of a chalcone derivative and a dehydroprenyl 2-arylbenzofuran derivative. Intravenous injection of mulberrofuran C produced significant hypotension in rabbit. PMID- 17396935 TI - Iridoids in equatorial and tropical flora. AB - The leaves of Tecoma chrysantha Jacq. a tree of the tropics, contain together with 6-epiaucubin ( 1), a less polar iridoid 2. Chemical and spectroscopical data demonstrated that 2 contains a residue of 1 esterified by 8-hydroxy-2,6-dimethyl 2,6-octadienoic acid ( 3), a rare monoterpenic product. PMID- 17396936 TI - 8-epiloganic acid and 7-beta-hydroxy-8-epiiridodial glucoside. AB - Two new iridoid glucosides, isolated from Linaria cymbalaria (Scrophulariaceae), are 8-epiloganic acid ( 1) and 7-beta-hydroxy-8-epiiridodial glucoside ( 2). Partial synthesis of 2 from 8-epiloganin ( 3) has been performed. PMID- 17396937 TI - [Oleo resin of Aucoumea klaineana]. PMID- 17396938 TI - Isolation of 6-O-(p-coumaroyl)-catalpol from Tabebuia rosea. AB - From the bark of Tabebuia rosea, the iridoid 6-O-(p-coumaroyl)-catalpol (specioside) was isolated. Antimalarial properties have been attributed to the infusion prepared from this part of the plant, although the compound failed to exhibit antimicrobial activity. The structure was corroborated by comparison with reported data for specioside as well as some additional chemical and spectroscopic data described herein. PMID- 17396939 TI - 8-Epi-iridodial glucoside from Boschniakia rossica. AB - Boschnaside, 8-epi-iridodial glucoside a new iridoid glucoside was isolated from Boschniakia rossica Hult., and its absolute structure was determined by chemical correlation of boschnaloside tetraacetate with boschnaside tetraacetate. PMID- 17396940 TI - New oligospirostanosides and oligofurostanosides from Asparagus adscendens roots. AB - The methanol extract of the defatted roots of Asparagus adscendens Roxb. yielded beta-sitosterol beta-D-glucoside, two new spirostanol glycosides (asparanin C and asparanin D) and two new furostanol glycosides (asparoside C and asparoside D). PMID- 17396941 TI - Isolation and structural elucidation of some sapogenols from Abrus cantoniensis. AB - After hydrolysis of the saponin fraction of the roots of Abrus cantoniensis, a new triterpenoid sapogenol, cantoniensistriol, was isolated together with sophoradiol, soyasapogenol B and soyasapogenol A. The structures of cantoniensistriol and sophoradiol were elucidated by spectroscopic methods. On the basis of the relation between cantoniensistriol and soyasapogenol A, the steric assignment of the C-21 and C-22 hydroxyl groups of the latter was discussed. PMID- 17396942 TI - [Di-C-glycosylflavones from Cerastium arvense ssp. arvense new for Caryophyllaceae.]. AB - The C-galactosyl-6 C-arabinosyl-8 apigenin or isocorymboside and a C-xylosyl-6 C arabinosyl-8 apigenin are isolated from the fresh whole plant, among many other flavonoids only present in small amounts. PMID- 17396943 TI - Alcaloides du Comularia camerunensis. AB - Column chromatography of the extract of C. Camerunensis stems and root barks afforded the known indole alkaloids, pleiocarpine, pleiocarpinine, eburnamine, isoeburnamine and eburnamenine. PMID- 17396944 TI - Free and esterified fatty acids of guar gum. AB - Guar gum consists essentially of the polysaccharide galactomannan. However, small amounts of various lipid soluble compounds, including long chain fatty acids, are also present. In this investigation free and esterified fatty acids of guar gum were analysed by GLC-MS. Palmitate (hexadecanoate), oleate (9-octadecenoate) and linoleate (9,12-octadecadienoate) were found to be the main components of the lipid fraction, but several long chain saturated acids and at least two 2-hydroxy derivatives of fatty acids are also present. PMID- 17396945 TI - Confirmation of new, high-mass saponins from Gleditsia japonica by field desorption mass spectrometry. AB - The molecular weights and complete sugar sequences have been determined in saponins carrying 3 to 8 free sugars. The results are in agreement with parallel chemical and instrumental analyses of the triterpenoid saponins derived from Gleditsia japonica. The capacity and limitations of field desorption mass spectrometry in structural elucidation of substituted oligoglycosides are described. PMID- 17396946 TI - Gleditsia saponins. AB - The chemical structure of gleditsia saponin C (GS-C), a major triterpenoid saponin of the fruit capsule of Gleditsia japonica Miq. (Leguminosae), comprised echinocystic acid, eight sugar units and two monoterpenyl groups ((+)2,6-dimethyl and (+) 2-hydroxymethyl-6-methyl-6-(S)hydroxy-2- trans-2,7-octadienoyl groups). Methylation analysis of GS-C followed by determination with gas-liquid chromatography and mass spectrometry showed that the monoterpenyl moieties attach to the 2 and 3-positions of terminal L-rhamnose of the C-28 oligoside portion. PMID- 17396947 TI - Differentiation of alkaloid cells in cultures of Macleaya mesophyll protoplasts. AB - In developing cell clusters derived from mesophyll protoplasts of Macleaya cordata (Willd.) R. Br. and M. microcarpa ( Maxim.) Fedde (Papaveraceae) cytodifferentiation is observed in the form of alkaloid cells and tracheary elements. The differentiation of alkaloid cells can already occur at the early stage of small clusters. Moreover cell colonies have been observed consisting predominantly to completely of alkaloid cells. Occasionally the direct differentiation of alkaloid cells is noticed, characterized by the coloured contents of the vacuoles after cell wall regeneration, but without preceding cell division. PMID- 17396948 TI - Cryopreservation of Digitalis lanata cell cultures. AB - Suspension cultures of Digitalis lanata strain I were grown in a medium containing 3% mannitol. For cryopreservation cell suspensions were treated with a mixture of sucrose-glycerol (20%/20 V%), cooled slowly (about 1 degrees C/min) till -100 degrees C and then were transferred to liquid nitrogen. After storage in liquid nitrogen the cells were thawed rapidly in a water bath of 40 degrees C and spread on the surface of a solidified nutrient medium. After 7 days of regrowth the cells were suspended in liquid nutrient medium for further cultivation. About 50% of the cells survived freezing and thawing. However, also the apparently surviving cells showed signs of injury (membrane vesicles outside the plasmalemma, dilated ER cisternae and separation of the nuclear membranes). The cultures derived from the surviving cells had the same growth rate and biochemical activity relative to the transformation of cardenolides, e.g., digitoxin, as the parent cultures. The frequency distribution of the nuclear DNA content in the cell cultures was the same before and after cryopreservation. These results indicate that there is no selection of a special cell type during freezing and thawing. PMID- 17396949 TI - [12-Methoxy-17,18-dehydro-vincamine and 16-epi-isositsirikine, alkaloids from Tabernaemontana psorocarpa]. AB - Besides sweroside ( 1) two main alkaloids were isolated from twigs and leaves of Tabernaemontana psorocarpa and shown to be 12-methoxy-17,18-dehydrovincamine ( 2) and 16-epi-isositsirikine ( 3) respectively. PMID- 17396950 TI - [Chemical composition of the essential oil from melissa]. AB - Different oil-samples of Melissa officinalis L. were analysed by capillary GC/MS, using fused silica columns and E.I.-Mass-spectrometry. Comparing the observed mass-spectra with those of a spectral collection, 70 compounds of the oil were identified. Geranial, neral, 6-methyl-5-hepten-2-one, citronellal, geranyl acetate, beta-caryophyllene, and beta-caryophyllene-oxide comprise about 96%. The fingerprint of the capillary gas-chromatogramm permitted differentiation of the essential oil of Melissa officinalis, Cymbopogon winterianus and Nepeta cataria var. citriodora., as well as a standardisation of pharmaceutical preparations containing Melissa oil. PMID- 17396951 TI - [Triterpenes and steroids in seeds of Solanum dulcamara]. AB - In seeds of Solanum dulcamara L. the 4,4-dimethylsterols cycloartenol, lanosterol, cycloartanol, 24-dihydrolanosterol, 24-methylenecycloartanol, and 24 methylenelanost-8-en-3beta-ol, the 4alpha-methylsterols 31-norcycloartenol, 31 norlanosterol, 31-norlanost-8-en-3beta-ol, lophenol, cycloeucalenol, obtusifoliol, gramisterol, and citrostadienol and the sterols cholesterol, 24 methylenecholesterol, campesterol, isofucosterol, sitosterol, and stigmasterol were identified by GC and GC/MS. The biosynthesis of these compounds is discussed. Additionally the spirostanoles tigogenin and diosgenin were isolated. The N-analoges spirosolanoles soladulcidine and solasodine were identified by TLC. The seeds contain 28% oil, of which the fatty acid composition has been examined. The predominant fatty acids are linolic acid (18:2) and oleic acid (18:1). PMID- 17396952 TI - [Studies on Ruta graveolens ssp. divaricata]. AB - Parts of leaves, stems and roots of Ruta graveolens ssp. divaricata (Tenore) Gams were explanted on Linsmaier and Skoog (16) medium. All explants showed good callus growth under various culture conditions. With stem derived tissue cultures, the capability for differentiation and regeneration was examined in response to variations of media and phytohormonal composition. In darkness, cultures produced roots of various length depending on composition of the media. Under continuous illumination (1200 lux) cultures produced fully green shoot regenerates on various media though complete plants could not be regenerated. PMID- 17396953 TI - [Analytical 1H-NMR-study of some natural Rotameric procyanidins]. AB - The phenomenon of conformational isomers of naturally occuring dimeric B procyanidins has been studied systematically by the 270 MHz- (1)H-NMR spectra of the acetate derivatives and the observed data are presented as an additional analytical criterion. PMID- 17396954 TI - Bio-Transformation of (G-3H)-digitoxin and (G-3H)-digoxin by Digitalis purpurea. AB - The biotransformation of (G- (3)H) digitoxin and (G- (3)H) digoxin has been studied in a strain of Digitalis purpurea in which cardenolides of the B series were predominant. A direct method of introducing the tritiated glycosides into the plant is described and the conversions monitored at intervals during the subsequent 6 weeks. Conversion of labelled digitoxin to purpurea glycosides A and B, gitoxin and gitaloxin occurred. Of particular interest was the conversion of digitoxin to the digitalose - containing glycosides, strospeside and verodoxin. Digoxin, an exogenous glycoside to D. purpurea, was glycosylated to desacetyl lanatoside C; there was some conversion to B and E series glycosides but only minimal formation of A series cardenolides. PMID- 17396955 TI - Etude des hernandiacees. AB - The alkaloidal content of the leaves and stem barks of H. peltata has been studied; eleven alkaloids and three lignans are isolated from the leaves, eleven alkaloids from the stem barks. Thirteen different alkaloids are identified by means of the spectral analysis; twelve are aporphines and nor-aporphines. PMID- 17396957 TI - [Flavonoids of Artemisia campestris, ssp. glutinosa]. AB - Four flavanones (pinostrobin, pinocembrin, sakuranetin and naringenin), one dihydroflavonol (7-methyl aromadendrin) and one flavone (hispidulin) have been isolated from Artemisia campestris L. ssp. glutinosa Gay and identified by spectroscopic methods. Artemisia campestris L. sous-espece glutinosa Gay est une Composee Anthemidee largement repandue sur les sables du littoral mediterraneean et abondante dans certaines regions d'Espagne et d'Italie. Dans le cadre d'une etude chimiotaxonomique du genre Artemisia Tourn., nous nous sommes interesses a l'analyse des flavonoides, composes jamais decrits, a notre connaissance, dans cette espece d' Artemisia. Les sommites fleuries d' Artemisia campestris sous espece glutinosa, sechees et pulverisees, sont degraissees a l'ether de petrole et epuisees par le chloroforme. Le fractionnement de l'extrait chloroformique, par chromatographie sur colonne de silice, et la purification de certaines fractions conduisent a l'isolement de six genines flavoniques, a l'etat pur. L' etude des spectres UV, des spectres de masse et des spectres de RMN [1,2] et la comparaison avec des echantillons authentiques permettent de proposer, pour ces flavonoides, les structures de la pinostrobine [3], de la pinocembrine [4], de la sakuranetine, de la naringenine [5] (flavanones), de la methyl-7-aromadendrine, [6, 7] (dihydroflavonol) et de l'hispiduline [8, 9] (flavone); quatre de ces genines sont methylees. Parmi ces flavonoides, la pinostrobine n'a jamais ete decrite, a notre connaissance, dans la famille des Composees; la pinocembrine, la sakuranetine et la naringenine ont deja ete signalees chez quelques Asteracees et Eupatoriees [10], et l'hispiduline dans la tribu des Anthemidees ( Santolina chamaecyparissus L.) [8]. Seule, la methyl-7-aromadendrine semble decrite, a ce jour, dans le genre Artemisia Tourn. [7]. PMID- 17396958 TI - Pyrrolizidine alkaloids from Senecio cruentus. PMID- 17396959 TI - Enzymatic determination of total cardiac glycosides in plant extracts. PMID- 17396960 TI - [New results of ergot alkaloid research]. PMID- 17396961 TI - Phenolic glycosides of Paulownia tomentosa bark. AB - The isolation of acteoside and coniferin from Paulownia tomentosa bark along with the previously reported phenolic glucoside syringin is described. The structure of both, acteoside and coniferin, have been assigned by (1)H- and (13)C-NMR spectroscopy. PMID- 17396963 TI - [Triterpenes and steroids in callus cultures of Solanum dulcamara]. AB - From tissue cultures (callus) of the soladulcidine strain of Solanum dulcamara L. the 4,4-dimethylsterols cycloartenol, cycloartanol, 24-dihydrolanosterol, and 24 methylenecycloartanol and the sterols cholesterol, 24-methylenecholesterol, campesterol, stigmasterol, isofucosterol, and sitosterol were identified by GC and GC/MS. In the callus the sterols are present in the free form and as esters, glucosides and palmitic esters of the glucosides. The total contents and the contents of the combined forms were determined photometrically (total: 0.19% on dry weight basis, 59.4% glucosides and acylated glucosides, 4.7% esters, 35.9% free). The main fatty acids of the petrolether soluble lipids of the callus (1.78%) are palmitic-, linoleic-, and linolenic acid. PMID- 17396964 TI - [Physcion-8-O-beta-D-gentiobioside, a new anthraquinone glycoside from rhubarb roots]. AB - A new anthraquinone glycoside - physcion-8-O-beta-D-gentiobioside - has been isolated from rhubarb roots in high yield. The structure was confirmed by H-NMR and MS-data as well as by direct comparison with the synthesized compound. The new substance was the main anthraquinone glycoside in some of the investigated samples of rhubarb roots and extracts. PMID- 17396966 TI - Constituents of the cultivated mulberry tree. AB - From the ethyl acetate extract of root bark of the Japanese cultivated mulberry tree (a variety of Morus alba L.), a novel chalcone derivative with a fused dihydrochalcone partial moiety was isolated and named kuwanon I. The structure was shown to be I on the basis of chemical and spectral data. Kuwanon I (I) is the first example which is regarded biogenetically as a Diels-Alder adduct of a prenylchalcone derivative and a dehydroprenylchalcone derivative. NMR variable temperature studies of I suggested that kuwanon I (I) exists as an equilibrium mixture of conformational isomers in solution. PMID- 17396968 TI - Alkaloids of Turkish Glaucium species. AB - Six isoquinoline alkaloids, glaucine, isocorydine, protopine, cryptopine, allocryptopine and trans-canadine methochloride, have been obtained from the aerial parts of Glaucium grandiflorum Boiss. et Huet var. torquatum Cullen, a plant indigenous to Turkey. The presence of a seventh alkaloid, corydine, was revealed by TLC. This is the first report of the occurrence of cryptopine in the genus Glaucium. PMID- 17396967 TI - Alkaloids from Papaver cylindricum. AB - The major alkaloids obtained from Papaver cylindricum Cullen were oripavine, narcotine, thebaine and rhoeadine. Floripavidine, papaverine, salutaridine, armepavine, N-methylasimilobine, scoulerine and cheilanthifoline were obtained as minor alkaloids. There have been no previous reports of alkaloids from P. cylindricum. PMID- 17396969 TI - Molluscicidal properties of Acacia nilotica. AB - The acetone, alcohol and aqueous extracts of fruits of Acacia nilotica (L) Willd. ex Del. (Mimosaceae) showed molluscicidal activity against Bulinus truncatus and Biomphalaria pfeifferi, the two snail species which transmit schistosomiasis in the Sudan (S. haematobium and S. mansonii). TAN, a new vegetable molluscicide prepared from the aqueous extract of the fruits of A. nilotica was produced on a pilot scale by using the spray drying procedure. Due to the high content of tannins in the powder (more than 56%), the molluscicidal properties of this plant may well be due to these compounds. PMID- 17396970 TI - [Comparative study of two medicinal Cucurbitaceae]. AB - Comparative study of the seeds of two Cucurbitaceae Momordica charantia L. and Herpetospermum caudigerum Wall. shows and important difference for their metabolism concerning particularly aspartic acid. Momordica can produce vicin, while for Herpetospermum, this amino acid would give asparagin. In the other hand, between the two species, Herpetospermum is the one which is able to accumulate lignoids. PMID- 17396971 TI - [Indole alkaloids of Odyendea gabonensis]. AB - From the trunk bark of Odyendea gabonensis six indole alkaloids have been isolated: Canthin-6-one (I); 5-Methoxy-canthin-6-one (II); 4,5-Dimethoxycanthin-6 one (III); 8-Hydroxy-canthin-6-one (IV); 1-Hydroxy-methyl-beta-carboline (V); and 1-Carboxamide-beta-carboline (VI). This is the first report of these products from this plant. PMID- 17396972 TI - Preliminary investigation of Ectopopterys soejartoi. PMID- 17396973 TI - Hydroxyanthraquinones from Abrus cantoniensis. PMID- 17396974 TI - Plant molluscicides. PMID- 17396979 TI - Alkaloids from Chasmanthera dependens. AB - A phytochemical investigation of the stem of Chasmanthera dependens Hochst led to the isolation and identification of five quaternary alkaloids: jatrorrhizine, columbamine, pseudocolumbamine, magnoflorine and palmatine and nine tertiary non phenolic alkaloids: tetrahydropalmatine, liriodenine, lysicamine, oxoglaucine, glaucine, anonaine, nornuciferine, norglaucine and O,O-dimethylcorytuberine. PMID- 17396980 TI - [A comparative study of the effects of selection and environmental factors on alkaloid accumulation in strains of Choisya ternata]. AB - The accumulation of two dihydrofuroquinoline alkaloids has been studied in seventeen strains of Choisya ternata (Rutaceae) initiated from the same tree. The contents of platydesminium and balfourodinium vary in heavy proportions. The effect of some factors (age of the culture, NAA or glucose concentrations of the medium, light intensity) has been studied in two of these strains, one with a low accumulative capacity, the other with a high accumulative capacity. These factors may in certain cases increase the alkaloid contents of both strains, but the induced variability is much lower than that existing naturally between the strains. The strain with a low content always remains inferior to that with a high content. This shows the importance of selecting strains with a high accumulative capacity before studying an optimal production medium. PMID- 17396981 TI - Monoterpenes in the essential oil of Origanum majorana. AB - The essential oil of Origanum majorana L. plants, wild growing in Turkey, was investigated by means of LSC and GLC. The oil contained about 65% carvacrol and 4% thymol. PMID- 17396982 TI - A new flavone glycoside from seeds of Daucus carota. AB - Phytochemical examination of seeds of Daucus carota L. resulted in the isolation of apigenin -4'-O-beta-D-glucoside, kaempferol-3-O-beta-D-glucoside and a new flavone glycoside which was characterised as apigenin-7-O-beta-D-galactopyranosyl (1-->4)-O-beta-D-mannopyranoside. PMID- 17396984 TI - A new anthraquinone from Cassia sophera heartwood. AB - A novel anthraquinone, now named sopheranin, has been isolated from the heartwood of Cassia sophera and characterised as 1, 3, 6, 8-tetrahydroxy 2-methyl 7-vinyl anthraquinone along with (beta-sitosterol, chrysophanol, physcion and emodin. PMID- 17396985 TI - Phytochemical investigation of roots of Adhatoda vasica. PMID- 17396986 TI - Alkaloids of Senecio tenuifolius. PMID- 17396987 TI - Constituents of French turpentine of oil from Pinus pinaster (Bordeaux). PMID- 17396988 TI - [Two lipophilic flavonoids from Begonia glabra]. PMID- 17396989 TI - Lessons from an HIV transmission pair. PMID- 17396990 TI - Herpes simplex virus type 2 and HIV-1: the dialogue between the 2 organisms continues. PMID- 17396991 TI - Transmission of hepatitis B in the health care setting: the elephant in the room ... or the mouse? PMID- 17396992 TI - Adrenergic regulation of bacterial virulence. PMID- 17396993 TI - The probable source of both the primary multidrug-resistant (MDR) HIV-1 strain found in a patient with rapid progression to AIDS and a second recombinant MDR strain found in a chronically HIV-1-infected patient. AB - BACKGROUND: Rapid progression to AIDS after acute infection with a multidrug resistant (MDR), dual-tropic strain of human immunodeficiency virus type 1 (HIV 1) was reported in a New York City man (hereafter referred to as "NYC") who has sex with men. The probable source of this HIV-1 (hereafter referred to as "CT01") and the development of a recombinant MDR HIV-1 in the source's partner (hereafter referred to as "CT02") are described. METHODS: After identification of the epidemiological link of CT01 and CT02 to NYC, viral sequences and phenotypic analyses were compared. Confirmatory genotypic and phenotypic analyses, replicative capacity, and viral coreceptor use were assessed. Viral recombination was assessed using a sliding window technique and phylogenetic tree analysis. RESULTS: NYC and CT01 were linked historically and epidemiologically and were genetically confirmed from CT01's samples acquired 2 days before and subsequent to the transmission event. Genotypic, recombination, and phylogenetic analyses suggest CT02 became superinfected by CT01 with subsequent production of a recombinant panresistant HIV-1. CONCLUSION: The probable source of a dual-tropic, MDR HIV-1 that was associated with rapid progression to AIDS is illustrated, suggesting progression was not explained by the HIV-1 variant alone. A probable second finding of a chronically infected host becoming superinfected with MDR HIV 1 with subsequent formation of a panresistant recombinant HIV-1 is described. This case illustrates the public health implications of unsafe sex between serodiscordant and seroconcordant partners. PMID- 17396994 TI - The role of herpes simplex virus type 2 and other genital infections in the acquisition of HIV-1 among high-risk women in northern Tanzania. AB - BACKGROUND: We examined the role of herpes simplex virus type 2 (HSV-2) and other genital infections on human immunodeficiency virus type 1 (HIV-1) incidence in a cohort study conducted between 2002 and 2005 among female bar/hotel workers in Moshi, Tanzania. METHODS: At baseline and every 3 months thereafter, participants were interviewed, and blood and genital samples were collected. Predictors of HIV 1 incidence were evaluated using a Cox proportional hazards regression model. RESULTS: Of 845 women who were HIV-1 seronegative at baseline, 689 (81.5%) were monitored in the study for a total of 698.6 person-years at risk (PYARs). The overall HIV-1 incidence was 4.6/100 PYARs (95% confidence interval [CI], 3.0 6.2/100 PYARs), and condom use was very low. After adjustment for other risk factors, the risk of HIV-1 was increased among women with HSV-2 at baseline (hazard ratio [HR], 4.3 [95% CI, 1.5-12.4]) and in those who acquired HSV-2 during the study period (HR, 5.5 [95% CI, 1.2-25.4]). Other independent predictors of HIV-1 were baseline chlamydial infection (HR, 5.2), bacterial vaginosis (HR, 2.1), and the occurrence of genital ulcers (HR, 2.7). CONCLUSION: HSV-2 and other genital infections were the most important risk factors for HIV 1. Control of these infections could help to reduce HIV-1 incidence in this population. PMID- 17396995 TI - Herpes simplex virus type 2 infection does not influence viral dynamics during early HIV-1 infection. AB - OBJECTIVE: We sought to compare baseline and longitudinal plasma HIV-1 loads between herpes simplex virus type 2 (HSV-2)-seropositive and -seronegative individuals who are enrolled in a primary HIV-1 infection cohort in San Diego, California. DESIGN: The study was a retrospective cohort analysis. METHODS: We categorized antiretroviral-naive subjects on the basis of HSV-2 serostatus at baseline using an HSV-2 enzyme immunoassay. Low positive results (1.1-3.5) were confirmed by Western blotting. We compared baseline HIV-1 loads of the 2 groups using a linear model. To detect differences in HIV-1 dynamics, we analyzed longitudinal viral loads using a flexible semiparametric model, controlling for the time to antiretroviral therapy and stratifying by HIV-1 infection stage at entry. RESULTS: We studied 294 adult men. Ninety percent reported sex with men as their main HIV-1 risk factor. The seroprevalence of HSV-2 was 41.5%. The HSV-2 seropositive and -seronegative groups had similar baseline HIV-1 loads during acute infection (5.52 vs. 5.72 log(10) copies/mL; P=.39) and early infection (4.57 vs. 4.67 log(10) copies/mL; P=.5). Longitudinally, the difference in HIV-1 loads between HSV-2-seropositive and -seronegative men remained close to 0 during the first year of infection. CONCLUSIONS: HSV-2 serostatus has minimal influence on the dynamics of HIV-1 during acute and early HIV-1 infection. PMID- 17396996 TI - HIV and parasitic infection and the effect of treatment among adult outpatients in Malawi. AB - We measured enteric parasitic infection prevalence and the effect of treatment on human immunodeficiency virus (HIV) RNA levels to assess their importance to HIV primary care in resource-limited settings. Adults in Lilongwe, Malawi, were evaluated, treated, and followed-up for parasitic and HIV infections. Of 389 patients, 266 (68%) were HIV infected. Helminth infections were more common in HIV-uninfected than in HIV-infected patients (39% vs. 17%). Among HIV-infected patients, helminth infections were associated with higher CD4 cell counts but not with higher HIV RNA levels. Successful treatment of parasitic infections had no effect on HIV RNA levels. Although common, parasitic infections did not impact HIV RNA levels. PMID- 17396997 TI - Molecular epidemiology of HIV-1 infection and full-length genomic analysis of circulating recombinant form 07_BC strains from injection drug users in Taiwan. AB - BACKGROUND: Previously, we reported that there was an outbreak of human immunodeficiency virus type 1 (HIV-1) circulating recombinant form (CRF) 07_BC among injection drug users (IDUs) in Taiwan in 2004. The objectives of the present study were to conduct a molecular epidemiological analysis and to characterize the full-length genome of the Taiwanese CRF07_BC. METHODS: Three hundred and fifty-eight patients with HIV-1/AIDS from hospitals and 133 HIV-1 infected inmates from detention centers were recruited. DNA sequencing and phylogenetic analysis were conducted to determine subtypes and evolutionary relationships. Recombination breakpoints of 2 full-length CRF07_BC strains were elucidated using a bootscanning method. RESULTS: Of 206 HIV-1-infected patients who received a diagnosis in 2004, 44.7% were infected with subtype B, 53.4% with CRF07_BC, and 1.5% with CRF01_AE. Ninety-eight percent (109/111) of IDUs were infected with CRF07_BC. Deletions of 7-11 amino acids in both p6(gag) and p6(pol) proteins were noted among the Taiwanese CRF07_BC strains. The CRF_07BC strains belonged to 2 phylogenetic clusters, and the first cluster contained only CRF07_BC strains from the southern part of Taiwan. CONCLUSIONS: The Taiwanese CRF07_BC strains had 97% full-length sequence homology with the prototype from mainland China. CRF07_BC was first introduced into the southern region in 2002 and then spread to other regions in Taiwan in 2004. PMID- 17396998 TI - Progesterone inhibits HIV-1 replication in human trophoblast cells through inhibition of autocrine tumor necrosis factor secretion. AB - BACKGROUND: Progesterone levels are higher in placental barriers during pregnancy, but the effect of progesterone on human immunodeficiency virus type 1 (HIV-1) infection in placental cells has not been addressed. We hypothesize that progesterone may affect HIV infection. METHODS: Purified trophoblastic cells and trophoblastic cell lines were infected or transfected with HIV-1, and the effect of progesterone was analyzed. Viral replication was measured by viral p24 or viral load quantification. Nuclear factor kappa -B (NF- kappa B) or long terminal repeat (LTR)-dependent transcription was measured by luciferase assays. Expression of chemokine receptors was analyzed by flow cytometry. Tumor necrosis factor (TNF) messenger RNA was assessed by reverse-transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR. RESULTS: Progesterone inhibits HIV-1 replication in placental cells at the concentration found in the placental interface, at a postentry step, and does not affect cell surface expression of chemokine receptors. Progesterone did not inhibit basal or induced LTR transcription or NF- kappa B activation. TNF synthesis in placental cells is induced by HIV-1 infection that, in an autocrine manner, activates viral replication, because neutralizing anti-TNF antibodies block it. Progesterone inhibits the induction of TNF synthesis by viral infection and virus or gp-120 induced TNF transcription. CONCLUSION: Our results demonstrate that progesterone inhibits HIV-1 replication in placental cells by reducing TNF levels, which are required for optimal viral replication. PMID- 17396999 TI - Mechanisms of polymorphonuclear neutrophil-mediated induction of HIV-1 replication in macrophages during pulmonary tuberculosis. AB - BACKGROUND: Pulmonary tuberculosis (TB) can present with polymorphonuclear neutrophil (PMN)-predominant alveolitis. TB accelerates acquired immunodeficiency syndrome by increasing human immunodeficiency virus type 1 (HIV-1) replication and mutation in alveolar macrophages. A 16-kDa CCAAAT/enhancer-binding protein beta (C/EBP beta ) isoform is a strong transcriptional repressor of the HIV long terminal repeat (LTR) in resting alveolar macrophages, leading to latent viral infection; its expression is lost during TB, derepressing the HIV LTR. METHODS: Lung segments were sampled from HIV/Mycobacterium tuberculosis-coinfected patients by means of bronchoalveolar lavage. In vitro coculture experiments defined the mechanism of induction of HIV-1 infection in macrophages by PMNs. RESULTS: Lung segments from patients with PMN-predominant TB had a markedly elevated viral load. Direct contact between activated PMNs and macrophages stimulated HIV-1 replication and LTR transcription and down-regulated inhibitory C/EBP beta . Isolated PMN membranes substituted for PMN contact, derepressing the HIV-1 LTR. The lipid raft fraction of PMN membranes expressed CD40 ligand (CD40L), CD28, and leukocyte function-associated antigen 1 (LFA-1 [i.e., CD11a and CD18]), and PMN activation increased lipid raft expression of CD40L and CD28. Blocking antibodies to CD40L, CD28, and LFA-1 inhibited PMN membrane-mediated HIV 1 LTR derepression. Alternately, cross-linking of macrophage receptors for CD40L, CD28, and LFA-1 (CD40, CD80/86, and intercellular adhesion molecule 1) abolished inhibitory C/EBP beta expression. CONCLUSION: PMN-macrophage contact derepresses the HIV-1 LTR and enhances HIV-1 replication in alveolar macrophages during pulmonary TB. Derepression is mediated through costimulatory molecule signaling. PMID- 17397000 TI - Patient-to-patient transmission of hepatitis B virus associated with oral surgery. AB - We used molecular epidemiologic techniques to document patient-to-patient transmission of hepatitis B virus (HBV) between 2 outpatient oral surgery patients operated on 161 min apart. Serological testing of 25 (93%) of 27 patients operated on after the source patient revealed that 19 (76%) of 25 were previously immune to HBV; no additional cases were identified. We found no deficiencies in infection control practices. Transmission may have been limited by the high prevalence (64%) of patients vaccinated against HBV. To our knowledge, this is the first documented case of patient-to-patient transmission of a bloodborne pathogen in a dental setting in the United States. PMID- 17397001 TI - CD4+ T cell responses in patients with chronic hepatitis C undergoing peginterferon/ribavirin therapy correlate with faster, but not sustained, viral clearance. AB - T cell immune responses may be important for the elimination of chronic hepatitis C virus (HCV) infection during antiviral treatment. In the present study, the kinetics of T cell responses to HCV antigens (NS3-4 and core) were prospectively assessed and were correlated with virologic outcome in 31 patients with chronic HCV infection undergoing peginterferon- alpha 2a/ribavirin therapy. NS3-4- directed T helper cell type 1 (Th1) responses were detected in 77% of patients with a significant decline in viremia at treatment week 4 but were not detected not in those with a slower viral decline. The detectability of NS3-4--directed Th1 responses was associated with faster viremia clearance, was short-lived, and did not seem to be associated with the final treatment outcome. PMID- 17397002 TI - GSTT1 and GSTM1 null genotypes may facilitate hepatitis C virus infection becoming chronic. AB - Oxidative stress contributes to hepatitis C virus (HCV)-induced liver damage. The activity of antioxidant glutathione S-transferases (GSTs) T1 and M1 is polymorphic. The GSTT1 and GSTM1 genotypes were identified in 139 HCV-infected patients and in 329 healthy individuals. Among patients, there was an excess of GSTT1 (odds ratio [OR], 2.76 [95% confidence interval [CI], 1.77-4.30]; P<.001) and GSTM1 (OR, 1.54 [95% CI, 1.02-2.35]; P=.032) null genotypes and of double null haplotypes (OR, 3.65 [95% CI, 1.98-6.75]; P<.001). The GSTT1 null genotype, particularly if associated with the GSTM1 null genotype, may facilitate HCV infection becoming chronic. PMID- 17397003 TI - Topical resiquimod 0.01% gel decreases herpes simplex virus type 2 genital shedding: a randomized, controlled trial. AB - BACKGROUND: Resiquimod, an investigational immune response modifier and Toll-like receptor (TLR) 7 and 8 agonist, stimulates production of cytokines that promote an antigen-specific T helper type 1 (Th1)--acquired immune response. In animal models, induction of Th1-specific responses modifies experimental herpes simplex virus (HSV) infection. METHODS: We conducted a randomized, double-blind, vehicle controlled trial to assess the efficacy of resiquimod 0.01% gel for reducing human anogenital HSV-2 mucosal reactivation. Adults with genital HSV-2 applied resiquimod or vehicle topically to herpes lesions 2 times weekly for 3 weeks and then collected daily anogenital swabs for 60 days for HSV DNA polymerase chain reaction. Recurrences during the subsequent 7 months were treated with study gel. During the final treatment-free 60 days, participants again collected daily swabs to assess shedding. RESULTS: The median lesion and shedding rates were lower for resiquimod compared with vehicle recipients during the initial sampling period (10% vs. 16% [P=.03] and 10% vs. 17% [P=.08], respectively) and during the final sampling period (3% vs. 22% [P<.001] and 10% vs. 26% [P=.009], respectively). Resiquimod did not influence recurrence length. CONCLUSIONS: These findings suggest that the immunological control of HSV-2 reactivation and lesion clearance may differ and that TLR7 and TLR8 agonists can reduce the frequency of mucosal HSV-2 reactivation. PMID- 17397004 TI - A comprehensive murine model to evaluate topical vaginal microbicides: mucosal inflammation and susceptibility to genital herpes as surrogate markers of safety. AB - A critical gap in microbicide development is the absence of surrogate safety markers. The objective of the present study was to develop a murine model to examine the mucosal response to microbicides and to assess the functional implication of observed changes. Mice received 14 daily intravaginal doses of nonoxynol-9, PRO 2000, or placebo gel. Nonoxynol-9 induced an inflammatory response characterized by increases in levels of cytokines and chemokines, recruitment of neutrophils and monocytes into the genital tract, and activation of the transcription factors NF- kappa B and activator protein-1. Minimal inflammation was observed in response to 2% PRO 2000. Nonoxynol-9-treated mice were significantly more susceptible to challenge with a low dose of herpes simplex virus type 2; the response of PRO 2000-treated mice was similar to the response to placebo. These findings suggest that PRO 2000 has little deleterious effect on mucosal immunity and, if validated by clinical experiences, support the inclusion of this model in the preclinical evaluation of future candidate microbicides. PMID- 17397005 TI - Risk of cervical intraepithelial neoplasia grade 2 or 3 after loop electrosurgical excision procedure associated with human papillomavirus type 16 variants. AB - Identification of factors associated with risk of relapse after treatment for high-grade cervical intraepithelial neoplasia (CIN) has important clinical implications. Study subjects were women participating in the Atypical Squamous Cells of Undetermined Significance and Low-Grade Squamous Intraepithelial Lesion Triage Study who were treated for CIN3 by loop electrosurgical excision procedure (LEEP) and who had a baseline infection with human papillomavirus type 16 (HPV16). These women were followed every 6 months for 2 years. Post-LEEP CIN2-3 was found in 20 (10.0%) of the 201 women. An adjusted relative risk of 3.1 (95% confidence interval, 1.1-8.9) was associated with HPV16 non-European, compared with European, variants, a finding that is consistent with the variant-related risk of prevalent/incident high-grade CIN. PMID- 17397006 TI - Comparison of tissue distribution, persistence, and molecular epidemiology of parvovirus B19 and novel human parvoviruses PARV4 and human bocavirus. AB - BACKGROUND: PARV4 and human bocavirus (HBoV) are newly discovered human parvoviruses with poorly understood epidemiologies and disease associations. We investigated the frequencies of persistence, tissue distribution, and influence of immunosuppression on replication of these viruses. METHODS: At autopsy, bone marrow, lymphoid tissue, and brain tissue from human immunodeficiency virus (HIV) infected individuals with acquired immunodeficiency syndrome (AIDS) and those without AIDS and from HIV-uninfected individuals were screened for parvovirus B19, PARV4, and HBoV DNA by means of quantitative polymerase chain reaction analyses. RESULTS: B19 DNA was detected both in HIV-infected study subjects (13 of 24) and in HIV-uninfected study subjects (8 of 8), whereas PARV4 DNA was detected only in HIV-infected study subjects (17 of 24). HBoV DNA was not detected in any study subjects. The degree of immunosuppression with HIV infection did not influence B19 or PARV4 viral loads. B19 or PARV4 plasma viremia was not detected in any study subjects (n=76; viral load <25 DNA copies/mL). A significantly older age distribution was found for study subjects infected with B19 genotype 2, compared with those infected with B19 genotype 1. Two genotypes of PARV4 were detected; study subjects carrying prototype PARV4 (genotype 1) were younger (all born after 1958) than those infected with genotype 2 (PARV5; study subjects born between 1949 and 1956). CONCLUSIONS: Tight immune control of replication of B19 and PARV4 was retained despite profound immunosuppression. Recent genotype replacement of PARV4, combined with absent sequence diversity among genotype 1 sequences, suggests a recent, epidemic spread in the United Kingdom, potentially through transmission routes shared by HIV. PMID- 17397007 TI - Modulation of pathogenicity with norepinephrine related to the type III secretion system of Vibrio parahaemolyticus. AB - BACKGROUND: Norepinephrine (NE) controls the functions of the gastrointestinal tract, but its role in the pathogenicity of enteropathogens is not clear. We examined the effect of NE on the pathogenicity of Vibrio parahaemolyticus with regard to its type III secretion systems (TTSSs). METHODS: To evaluate the effect of NE on pathogenicity of V. parahaemolyticus, we examined the cytotoxic activity to Caco-2 cells and enterotoxicity by use of the rat ileal loop model. It has been reported that TTSS1 causes cytotoxicity and that TTSS2 causes enterotoxicity in the animal ileal loop model. RESULTS: Our results showed that, although NE alone did not affect the viability of Caco-2 cells, NE stimulated the cytotoxic activity of V. parahaemolyticus. Furthermore, NE increased the transcription of the TTSS1-related genes vscQ and vscU. These results indicate that NE regulates V. parahaemolyticus cytotoxic activity. The enterotoxicity of V. parahaemolyticus was increased by NE through interaction with alpha (1)-adrenergic receptors. These results indicate that alpha (1)-adrenergic receptors on the intestinal epithelium appear to interact with V. parahaemolyticus enterotoxicity. CONCLUSIONS: The present findings suggest that enteric NE may modulate V. parahaemolyticus pathogenicity. PMID- 17397008 TI - Individuals with pulmonary tuberculosis have lower levels of circulating CD1d restricted NKT cells. AB - Mycobacterium tuberculosis (MTB) is a leading cause of mortality worldwide from an infectious agent. Natural killer T (NKT) cells recognize mycobacterial antigens and contribute to anti-MTB immunity in mouse models. NKT cells were measured in subjects with pulmonary tuberculosis, MTB-exposed individuals, and healthy controls. NKT cell levels are selectively lower in peripheral blood mononuclear cells from individuals with pulmonary tuberculosis than in both MTB exposed subjects and healthy control subjects. This apparent loss of NKT cells from the peripheral blood is sustained during the 6 months after the initiation of MTB treatment. These findings indicate that NKT cells may be an important component of antituberculosis immunity. PMID- 17397009 TI - Staphylococcus aureus exploits cathelicidin antimicrobial peptides produced during early pneumonia to promote staphylokinase-dependent fibrinolysis. AB - The increasing prevalence of Staphylococcus aureus strains isolated from hospital and community-acquired respiratory tract infections is an important public health concern worldwide. The majority of S. aureus strains produce staphylokinase, a plasminogen activator capable of inactivating neutrophil alpha defensins and of impairing phagocytosis via opsonin degradation. Cathelicidin antimicrobial peptides are present at sites of infection before the release of neutrophil alpha-defensins. Therefore, we hypothesized that staphylokinase interacts with cathelicidin during the early pathogenesis of S. aureus airway infection. In a mouse intranasal infection model, cathelicidin was strongly up regulated in the airways during the development of staphylococcal pneumonia. In vitro, cathelicidin bound directly to staphylokinase and augmented staphylokinase dependent plasminogen activation and fibrinolysis at concentrations consistent with those detected in the airways during infection. These data suggest that staphylokinase production may be a novel virulence mechanism by which S. aureus exploits cathelicidin to promote fibrinolysis, leading to enhanced bacterial dissemination and invasive infection. PMID- 17397010 TI - HLA class I-restricted T cell epitopes of the kinetoplastid membrane protein-11 presented by Leishmania donovani-infected human macrophages. AB - Visceral leishmaniasis is a protozoal disease caused by the intracellular parasites Leishmania donovani and L. chagasi/infantum, and it is usually deadly if not treated. To date, no vaccine exists for prophylaxis or immunotherapy, nor has it been established which effector mechanisms of the immune system are most instrumental against the parasites. Recent reports have suggested that CD8(+) T cells, in addition to CD4(+) T cells, might play major roles in the defense against infection and in the cure of the disease. To identify epitopes recognized by CD8(+) T cells that can be used for immune monitoring to investigate the role of these cells in human visceral leishmaniasis, as well as in vaccine development, we scanned the entire sequence of the leishmanial protein kinetoplastid membrane protein (kmp)-11 with overlapping nonapeptides. Thirty peptides that specifically trigger interferon- gamma secretion by human CD8(+) T cells were identified. Four T cell lines with specificities for different peptides recognize Leishmania-infected autologous macrophages, which proves that kmp-11 is processed and presented via the major histocompatibility complex class I pathway of infected cells. Kmp-11 is thus a candidate antigen for the development of T cell vaccines. PMID- 17397011 TI - Multilocus polymerase chain reaction restriction fragment--length polymorphism genotyping of Trypanosoma cruzi (Chagas disease): taxonomic and clinical applications. AB - BACKGROUND: Trypanosoma cruzi, the agent of Chagas disease, is subdivided into 6 discrete typing units (DTUs); their identification is important to understand clinical pleomorphism and track sylvatic DTUs that might (re-)invade domestic foci of the disease and jeopardize the running control programs. METHODS: The genetic polymorphism of 12 loci was analyzed by multilocus polymerase chain reaction restriction fragment--length polymorphism (PCR-RFLP) analysis (MLP analysis) in a sample representative of the diversity within T. cruzi. We paid particular attention to genes involved in host-parasite relationships, because these may be prone to polymorphism as an adaptive answer to the immune selective pressure. RESULTS: The results of MLP analysis were shown to agree with the current multilocus enzyme electrophoresis- and random amplified polymorphic DNA based classification of T. cruzi in 6 DTUs, thereby providing a taxonomic validation of our method. Our data supported hypotheses of genetic recombination within T. cruzi. We demonstrated direct applicability of PCR-RFLP analysis to blood of mammal hosts and intestine content of vector insects. Domestic DTUs were encountered in wild animals, and, reciprocally, sylvatic DTUs were encountered in humans, raising questions about changes of transmission patterns. CONCLUSIONS: MLP analysis represents a new alternative to existing molecular methods for T. cruzi typing. It might offer an invaluable support to clinical and epidemiological studies and to control programs. PMID- 17397012 TI - Unlikely association of multidrug-resistance protein 2 single-nucleotide polymorphisms with tenofovir-induced renal adverse events. PMID- 17397015 TI - On recombinant human erythropoietin preventing the death of mice during cerebral malaria. PMID- 17397016 TI - Direct EPR detection of a hydrogen-bonded complex between a semiquinone radical anion and a protonated amino acid, and electron transfer driven by hydrogen bonding. PMID- 17397017 TI - A supramolecular ferroelectric realized by collective proton transfer. PMID- 17397018 TI - Fast electrochemically induced translation of the ring in a copper-complexed [2]rotaxane: the biisoquinoline effect. PMID- 17397019 TI - High-yield synthesis of medium and large platinacycloalkanes from bis(alkenyl) precursors. PMID- 17397020 TI - Tuning electron transfer through translational motion in molecular shuttles. PMID- 17397021 TI - Metalloenzyme-inspired catalysis: selective oxidation of primary alcohols with an iridium-aminyl-radical complex. PMID- 17397022 TI - Stabilization of the zwitterionic structure of proline by an alkylammonium ion in the gas phase. PMID- 17397023 TI - Recognition of DNA three-way junctions by metallosupramolecular cylinders: gel electrophoresis studies. AB - The interaction of metallosupramolecular cylinders with DNA three-way junctions has been studied by gel electrophoresis. A recent X-ray crystal structure of a palindromic oligonucleotide forming part of a complex with such a cylinder revealed binding at the heart of a three-way junction structure. The studies reported herein confirm that this is not solely an artefact of crystallisation and reveal that this is a potentially very powerful new mode of DNA recognition with wide scope. The cylinders are much more effective at stabilizing three-way junctions than simple magnesium di-cations or organic or metallo-organic tetra cations, with the M cylinder enantiomer being more effective than P. The recognition is not restricted to three-way junctions formed from palindromic DNA with a central AT step at the junction; non-palindromic three-way junctions and those with GC steps are also stabilised. The cylinder is also revealed to stabilise other Y-shaped junctions, such as that formed at a fraying point in duplex DNA (for example, a replication fork), and other DNA three-way junction structures, such as those containing unpaired nucleotides, perhaps by opening up this structure to access the central cavity. PMID- 17397024 TI - Design and generation of extended zeolitic metal-organic frameworks (ZMOFs): synthesis and crystal structures of zinc(II) imidazolate polymers with zeolitic topologies. AB - Attempts to create metal-organic frameworks (MOFs) with zeolitic topologies, metal (zinc(II) and cobalt(II)) imidazolates have repeatedly been used as the metal-organic motifs of inorganic silicate analogues. By modulating the synthetic strategy based on the solvothermal and liquid diffusion method, seven further MOFs (including at least three zeolitic MOFs) of zinc(II) imidazolates, [Zn(im)2.x G] (G=guest molecule, x=0.2-1) 1 a-7 a, have been successfully synthesized. Of these, 1 a-3 a are isostructural with the previously reported cobalt analogues 1 b-3 b, respectively, while 4 a-7 a are new members of the metal imidazolate MOF family. Complex 4 a exhibits a structure related to silicate CaAl2Si2O8 of CrB4 topology, but with a higher network symmetry; complex 5 a has a structure with zeolitic DFT topology that was discovered in zeolite related materials of DAF-2, UCSB-3, and UCSB-3GaGe; complex 6 a demonstrates an unprecedented zeolite-like topology with one dimensional channels with 10-rings; and 7 a displays a structure of natural zeolite GIS (gismondine) topology. All of these polymorphous MOFs were created only by using certain solvents as structure directing agents (SDAs). Further extensive metal-organic frameworks with zeolitic topologies can be envisaged if other solvents were to be used. PMID- 17397025 TI - Absolute configuration of conformationally flexible cis-dihydrodiol metabolites by the method of confrontation of experimental and calculated electronic CD spectra and optical rotations. AB - We have determined the absolute configurations of conformationally flexible cis dihydrodiol metabolites (cis-1,2-dihydroxy-3,5-cyclohexadienes), bearing different substituents (e.g., Br, F, CF3, CN, Me) in 3- and 5-positions, by the method of confrontation of experimental and calculated electronic CD spectra and optical rotations. Convergent results were obtained by both methods in eight out of ten cases. For the difficult cases, where either conformer population and/or chiroptical properties (calculated rotational strengths of the long-wavelength Cotton effect or optical rotations) of contributing conformers remain inconclusive, the absolute configuration could still be correctly assigned based on one of the biased properties (either ECD or optical rotation). This approach appears well-suited for a broad spectrum of conformationally flexible chiral molecules. PMID- 17397026 TI - Signs of proinflammatory/genotoxic switch (adipogenotoxicosis) in mammary fat of breast cancer patients: role of menopausal status, estrogens and hyperglycemia. AB - The abundance of fat tissue surrounding normal and malignant epithelial mammary cells raises the questions whether such "adipose milieu" is important in the local proinflammatory/genotoxic shift, which apparently promotes tumor development and worsens prognosis, and what conditions stimulate this shift, or "adipogenotoxicosis." We studied 95 mammary fat samples from 70 postmenopausal and 25 premenopausal breast cancer (BC) patients at a distance of 1.5-2.0 cm from tumors. The levels of leptin, adiponectin, TNFalpha and IL-6 release after 4-hr incubation of the samples were evaluated with ELISA, nitric oxide (NO) production by Griess reaction and lipid peroxidation by determination of thiobarbiturate reactive products (TBRP). Infiltration of fat with macrophages (CD68-positive cells) and expression of cytochrome P450 1B1/estrogen 4-hydroxylase (CYP1B1) were detected by immunohistochemistry. Aromatase (CYP19) activity in mammary fat was measured by (3)H(2)O release from (3)H-1beta-androstenedione. In the postmenopausal BC patients, NO and TNFalpha production by adipose tissue explants increased independent of BMI and in parallel with decreasing leptin and, especially, adiponectin release. In the premenopausal patients, higher CYP1B1 expression and TBRP level were found in mammary fat, while higher aromatase activity was combined with higher CYP1B1 expression as well as NO and IL-6 production. In the postmenopausal group, impaired glucose tolerance was associated with higher IL-6 release production by fat and with higher IL 6/adiponectin ratio. Thus, signs of adipogenotoxicosis in mammary fat can be found in both pre- and postmenopausal BC patients. This condition is likely being maintained through estrogen- and glucose-related factors and mechanisms presumably associated with less favorable types of hormonal carcinogenesis. PMID- 17397027 TI - Tumor-targeting Salmonella typhimurium improves cyclophosphamide chemotherapy at maximum tolerated dose and low-dose metronomic regimens in a murine melanoma model. AB - Chemotherapy for cancer is partly limited by the inability of drugs to act on poorly vascularized or avascularized areas of tumors. Tumor-targeting bacteria are capable of preferentially replicating in these poorly perfused regions. Some strains have been combined with chemotherapeutic agents and the results have been promising. However, no systematic work has been carried out to test the effect of bacteria on clinical modes of chemotherapy, such as standard maximum tolerated dose (MTD) and novel low-dose metronomic (LDM) chemotherapy. Here Salmonella typhimurium VNP20009 was combined with cyclophosphamide (CTX) at both MTD and LDM schedules in a murine melanoma model. The results showed that VNP20009 significantly improved the effects of all forms of CTX treatments. The combination of VNP20009 and CTX led to a more significant decrease in tumor microvessel density and serum vascular endothelial growth factor (VEGF) level, compared with either treatment alone. Furthermore, combination therapy remarkably increased the number of bacteria within tumors when compared with bacteria treatment alone. These findings suggest that tumor-targeting bacteria, in conjunction with CTX at standard MTD and LDM regimens, might be of clinical value for the treatment of melanoma. PMID- 17397028 TI - Identification of cytotoxic T-lymphocyte epitope(s) and its agonist epitope(s) of a novel target for vaccine therapy (PAGE4). AB - PAGE4 is an X chromosome-linked cancer testis antigen and is a potential new tumor-associated antigen that is overexpressed in prostate and uterine cancers. The purpose of this study was to identify a human CTL epitope and a corresponding agonist epitope of PAGE4 to determine if PAGE4 is a potential target for vaccine mediated immunotherapy against PAGE4-expressing tumors. A class I PAGE4 epitope was identified with a high level of binding to HLA-A2. PAGE4 peptide-pulsed dendritic cells were then used to generate human PAGE4-specific T-cell lines. Further studies demonstrated the generation of an enhancer agonist epitope. Compared with the native peptide, the agonist (i) bound to HLA-A2 molecules at lower peptide concentrations, (ii) demonstrated a higher stability of the peptide HLA-A2 complex, (iii) induced higher levels of production of IFN-gamma, Granzyme B, TNF-alpha, IL-2 and lymphotactin by PAGE4-specific T-cell lines and (iv) T cell lines generated against the agonist peptide were more efficient to lyse HLA A2 human tumor cells expressing native PAGE4. The studies reported here are the first to describe a PAGE4 CTL epitope and its agonist epitope, and thus identify PAGE4 as a potentially useful target for vaccine-mediated therapy of prostate cancer. PMID- 17397029 TI - Benefit of sentinel lymphadenectomy for patients with nonulcerated cutaneous melanomas in the Breslow range between 0.76 and 1 mm: a follow-up study of 148 patients. AB - Sentinel lymphadenectomy (SLNE) is now internationally accepted for the primary treatment of melanomas thicker than 1 mm. But it is still controversial whether also patients with nonulcerated melanomas in the Breslow range between 0.76 and 1 mm should be included. At the authors' department, 87 of such patients (Group A) underwent SLNE in combination with wide local excision (WLE) of their primary melanomas in the years 1995 to 2001. SLN micrometastases were found in 10 of these patients (11.5%). Radical completion lymph node dissections (CLND) were added in 4 of the 10 patients without revealing any further nodal metastases. All the 87 Group A patients remained free from recurrent disease at a median follow up time of 74 months. The control Group B from the same department encompassed 61 consecutive stage Ia patients with melanomas in the identical Breslow range, who had undergone only WLE of their primaries without SLNE in the years 1987 to 1993 (median follow-up time 115 months). Five of these 61 patients (8.2%) developed melanoma metastases within 12 to 68 (median 19) months of follow-up, 3 of them initially in regional lymph nodes. Four of the 5 individuals died because of the final distant dissemination of the melanoma. Kaplan-Meier comparisons between Groups A and B with log-rank testing showed a significantly worse outcome of Group B with respect to recurrence-free survival (p = 0.01), regional nodal progression (p = 0.041), distant metastasis (p = 0.023) and melanoma-related mortality (p = 0.03). The overall survival was not significantly different, because expiries not related to melanoma predominated in both groups. Our data suggest that SLNE seems to nearly completely eliminate the risk of melanoma recurrences in patients with melanomas between 0.76 and 1 mm thick. PMID- 17397030 TI - Frequent inactivation of SPARC by promoter hypermethylation in colon cancers. AB - Epigenetic modification of gene expression plays an important role in the development of human cancers. The inactivation of SPARC through CpG island methylation was studied in colon cancers using oligonucleotide microarray analysis and methylation specific PCR (MSP). Gene expression of 7 colon cancer cell lines was evaluated before and after treatment with the demethylating agent 5-aza-2'-deoxycytidine (5Aza-dC) by oligonucleotide microarray analysis. Expression of SPARC was further examined in colon cancer cell lines and primary colorectal cancers, and the methylation status of the SPARC promoter was determined by MSP. SPARC expression was undetectable in 5 of 7 (71%) colorectal cancer cell lines. Induction of SPARC was demonstrated after treatment with the demethylating agent 5Aza-dC in 5 of the 7 cell lines. We examined the methylation status of the CpG island of SPARC in 7 colon cancer cell lines and in 20 test set of colon cancer tissues. MSP demonstrated hypermethylation of the CpG island of SPARC in 6 of 7 cell lines and in all 20 primary colon cancers, when compared with only 3 of 20 normal colon mucosa. Immunohistochemical analysis showed that SPARC expression was downregulated or absent in 17 of 20 colon cancers. A survival analysis of 292 validation set of colorectal carcinoma patients revealed a poorer prognosis for patients lacking SPARC expression than for patients with normal SPARC expression (56.79% vs. 75.83% 5-year survival rate, p = 0.0014). The results indicate that epigenetic gene silencing of SPARC is frequent in colon cancers, and that inactivation of SPARC is related to rapid progression of colon cancers. PMID- 17397031 TI - Cutaneous phenotype and MC1R variants as modifying factors for the development of melanoma in CDKN2A G101W mutation carriers from 4 countries. AB - The G101W founder mutation is the most common CDKN2A mutation in Italy, Spain, and France. As the background of modifying genes, environmental exposures, and sun behavior vary across countries, studying G101W carriers from distinct countries offers a unique opportunity to evaluate possible modifying factors in melanoma development. We evaluated 76 G101W cases and 59 carrier controls from France, Italy, Spain, and the United States. Hair color and dysplastic nevi distributions differed significantly in cases and controls across the 4 study groups. Cases also varied significantly for eye color, freckling, and nevi. The distribution of MC1R variants in cases differed significantly across study groups because 12% of Italian melanoma patients had > or =2 MC1R variants vs. >50% for the other case groups. Several MC1R covariates showed significant associations with melanoma risk in all groups combined and in the American, French, and Spanish samples; no significant findings were observed in the Italian sample. In multiple-case families, the number and type of MC1R variants varied significantly between multiple-primary-melanoma and single-primary-melanoma patients from the 4 groups; there was also a significant decrease in median age at melanoma diagnosis as the number or type of MC1R variants increased. The variation in the effects of the cutaneous phenotypic and MC1R factors across the study sample suggests that these factors differentially contribute to development of melanoma even on a common genetic background of a germline CDKN2A mutation. Differences in melanoma risk across geographic regions justify the need for individual studies in each country before counseling should be considered. PMID- 17397032 TI - Diabetes mellitus and risk of breast cancer: a meta-analysis. AB - Diabetes mellitus has been associated with an increased risk of several types of cancers, but its relationship with breast cancer remains unclear. We conducted a meta-analysis of case-control and cohort studies to assess the evidence regarding the association between diabetes and risk of breast cancer. Studies were identified by searching MEDLINE (1966-February 2007) and the references of retrieved articles. We identified 20 studies (5 case-control and 15 cohort studies) that reported relative risk (RR) estimates (odds ratio, rate ratio/hazard ratio, or standardized incidence ratio) with 95% confidence intervals (CIs) for the relation between diabetes (largely Type II diabetes) and breast cancer incidence. Summary RRs were calculated using a random-effects model. Analysis of all 20 studies showed that women with (versus without) diabetes had a statistically significant 20% increased risk of breast cancer (RR, 1.20; 95% CI, 1.12-1.28). The summary estimates were similar for case-control studies (RR, 1.18; 95% CI, 1.05-1.32) and cohort studies (RR, 1.20; 95% CI, 1.11 1.30). Meta-analysis of 5 cohort studies on diabetes and mortality from breast cancer yielded a summary RR of 1.24 (95% CI, 0.95-1.62) for women with (versus without) diabetes. Findings from this meta-analysis indicate that diabetes is associated with an increased risk of breast cancer. PMID- 17397033 TI - Short- vs long-term androgen suppression plus external beam radiation therapy and survival in men of advanced age with node-negative high-risk adenocarcinoma of the prostate. AB - BACKGROUND: The study evaluated whether the use of 3 years as compared with 6 months of androgen suppression therapy (AST) combined with external beam radiation therapy (RT) in the treatment of high-risk prostate cancer was associated with prolonged survival in advanced age men. METHODS: A pooled analysis of 311 men enrolled in 3 prospective randomized trials between 1987 and 2000 who received 6 months or 3 years of AST and RT for locally advanced or high grade localized adenocarcinoma of the prostate comprised the study cohort. Cox regression multivariable analysis was performed adjusting for known prognostic factors to determine whether the treatment received was associated with time to death after randomization. The median age and follow-up was 70 and 5.9 years, respectively, during which 82 (26%) deaths occurred. RESULTS: Treatment received was not significantly associated with survival time after randomization (adjusted hazard ratio [AHR]: 1.1; 95% confidence interval [CI]: 0.7, 1.8; P = .70), whereas age at randomization (AHR: 1.05; 95% CI: 1.01, 1.09; P = .02) was. The presence of Gleason score 8 to 10 cancers approached significance (AHR: 1.6; 95% CI: 0.9, 2.6; P = .09). CONCLUSIONS: After adjusting for known prognostic factors, the treatment of node-negative, high-risk prostate cancer using 3 years as compared with 6 months of AST with RT was not associated with prolonged survival in men of advanced age. The European Organization for Research and Treatment of Cancer randomized trial will help answer whether unknown confounding factors affected the results of the study. PMID- 17397034 TI - Synthesis of porphyrin dimers fused with a benzene unit. AB - Bicyclo[2.2.2]octadiene-connected pyrrolo-porphyrins have been prepared by an inverse-type [3+1] porphyrin synthesis of a bicyclo[2.2.2]octadiene-fused dipyrrole with a tripyrrane dicarbaldehyde. Another [3+1] porphyrin synthesis of pyrrole-connected porphyrins with the same or other tripyrrane dicarbaldehydes gave bicyclo[2.2.2]octadiene-bridged diporphyrins, the central metals and/or peripheral substituents of which were different. Thermal decomposition of the bicyclo[2.2.2]octadiene skeleton to a benzene moiety gave pi-system-fused porphyrin dimers in a highly pure form. PMID- 17397035 TI - The impact on survival of thromboembolic phenomena occurring before and during protocol chemotherapy in patients with advanced gastroesophageal adenocarcinoma. AB - BACKGROUND: Thromboembolic events (TEEs) are considered common in patients with gastroesophageal carcinoma, but their frequency at baseline and during chemotherapy is not known. Because prophylactic anticoagulation results in improved overall survival (OS) of solid tumor patients, the authors hypothesized that TEEs at baseline and during chemotherapy would have an adverse effect on OS. METHODS: The authors analyzed patients with advanced gastroesophageal carcinoma who were treated on 4 prospective chemotherapy Phase II/III trials. Baseline and subsequent TEEs were documented and correlated with OS. RESULTS: On the 4 trials, 191 patients received single-agent or a combination of a taxane, camptothecin, platinum, or fluoropyrimidine. At baseline, TEEs occurred in 5.3% of untreated patients compared with 8.5% of previously treated patients (who had received prior treatment for metastatic disease). The median OS was only 3.9 months for patients who had a TEE at any time versus 8.7 months for patients who never developed a TEE (P = .007). TEEs at baseline were correlated with poor median OS in untreated patients (4.9 months vs 8.9 months for patients without a TEE; P = .014). There was no associated between TEEs and the type of chemotherapy used. CONCLUSIONS: The current results established that TEEs at baseline and/or during chemotherapy are frequent and result in poor OS for patients with advanced gastroesophageal carcinoma. Aggressive methods to treat or prevent TEEs are warranted. PMID- 17397036 TI - A cross-setting study of chimpanzee (Pan troglodytes) personality structure and development: zoological parks and Yerkes National Primate Research Center. AB - This study addressed whether personality ratings using a 43 adjective questionnaire based on the Five-Factor Model generalized from a sample of 202 zoo housed chimpanzees to a sample of 175 chimpanzees housed in Yerkes National Primate Research Center. Mean interrater reliabilities of adjectival ratings were lower for the chimpanzees housed in Yerkes. In addition, rank order of the interrater reliabilities of items differed between settings. To compare factor structure, we first examined whether we could replicate the original six factor structure found in an earlier study of 100 zoo chimpanzees using principal factors analysis in the Yerkes sample and 102 new zoo chimpanzees. The dominance, extraversion, conscientiousness, and agreeableness factors were clearly replicated in the Yerkes sample and the 102 new zoo chimpanzees. The Neuroticism and Openness factors did not replicate in the Yerkes sample, but they also did not replicate in the new zoo chimpanzees. These findings suggest the need to sample more adjectives representing neuroticism and openness in future versions of the questionnaire. We next sought to determine whether factor structure, as determined by principal components analysis, remained invariant across the two settings. This analysis revealed dominance, extraversion, conscientiousness, and agreeableness factors in both settings and a high level of congruence between the zoo and Yerkes samples for these factors. Finally, we tested whether factor scores in the two samples were similarly related to age and sex. With the exception of differences in age effects for dominance and agreeableness, age, and sex effects were consistent across samples. These findings suggest that, whereas there may be differences in the ease with which ratings are made, personality structure, and development are largely consistent across widely differing settings. PMID- 17397037 TI - Treatment with CRH-1 antagonist antalarmin reduces behavioral and endocrine responses to social stressors in marmosets (Callithrix kuhlii). AB - Corticotropin-releasing hormone (CRH) has multiple roles in coordinating the behavioral and endocrine responses to a host of environmental challenges, including social stressors. In the present study we evaluated the role of CRH in mediating responses to a moderate social stressor in Wied's black tufted-eared marmosets (Callithrix kuhlii). Male and female marmosets (n=14) were administered antalarmin (a selective CRH-1 receptor antagonist; 50 microg/kg, p.o.) or vehicle in a blind, counterbalanced, crossover design. One hr after treatment, marmosets were separated from long-term pairmates and then housed alone in a novel enclosure for 7 hr. Behavior was recorded during separation and upon reunion with the partner, and urine samples for cortisol assay collected before, during, and after the intervention. Separation from partners elevated urinary cortisol concentrations over baseline for both conditions, but antalarmin treatment reduced the magnitude of the elevation. Antalarmin also lowered rates of behavioral patterns associated with arousal (alarm and "e-e" vocalizations, object manipulate/chew), but had no effect on contact calls, locomotory activity or alertness. Although most patterns of social behavior upon reunion with the partner were not affected by antalarmin, antalarmin-treated marmosets displayed more sexual behavior (mounts and copulations) upon reunion. These data indicate that antagonism of the CRH-1 receptor acts to reduce the magnitude of both endocrine and behavioral responses to a moderate social stressor without causing any overall reduction in alertness or general activity. This supports the hypothesis that CRH, acting through its type 1 receptor, is involved in coordinating the responses to anxiety-producing events. These results further suggest that the marmoset is a useful model for exploration of the role of CRH in mediating the behavioral and neuroendocrine responses to psychosocial stressors, particularly in the context of heterosexual social relationships. PMID- 17397038 TI - Epistatic interactions with a common hypomorphic RET allele in syndromic Hirschsprung disease. AB - Hirschsprung disease (HSCR) stands as a model for genetic dissection of complex diseases. In this model, a major gene, RET, is involved in most if not all cases of isolated (i.e., nonsyndromic) HSCR, in conjunction with other autosomal susceptibility loci under a multiplicative model. HSCR susceptibility alleles can harbor either heterozygous coding sequence mutations or, more frequently, a polymorphism within intron 1, leading to a hypomorphic RET allele. On the other hand, about 30% of HSCR are syndromic. Hitherto, the disease causing gene has been identified for eight Mendelian syndromes with HSCR: congenital central hypoventilation (CCHS), Mowat-Wilson (MWS), Bardet-Biedl (BBS), Shah-Waardenburg (WS4), cartilage-hair-hypoplasia (CHH), Smith-Lemli-Opitz (SLO), Goldberg Sprintzsen (GSS), and hydrocephalus due to congenital stenosis of the aqueduct of sylvius (HSAS). According to the HSCR syndrome, the penetrance of HSCR trait varies from 5 to 70%. Trisomy 21 (T21) also predisposes to HSCR. We were able to collect a series of 393 patients affected by CCHS (n = 173), WS4 (n = 24), BBS (n = 51), MWS (n = 71), T21 (n = 46), and mental retardation (MR) with HSCR (n = 28). For each syndrome, we studied the RET locus in two subgroups of patients; i.e., with or without HSCR. We genotyped the RET locus in 393 patients among whom 195 had HSCR, and compared the distribution of alleles and genotypes within the two groups for each syndrome. RET acts as a modifier gene for the HSCR phenotype in patients with CCHS, BBS, and Down syndrome, but not in patients with MWS and WS4. The frequent, low penetrant, predisposing allele of the RET gene can be regarded as a risk factor for the HSCR phenotype in CCHS, BBS, and Down syndrome, while its role is not significant in MWS and WS4. These data highlight the pivotal role of the RET gene in both isolated and syndromic HSCR. PMID- 17397039 TI - The diffusion process of sodium hyaluronate (Na-Ha) and Na-Ha-n-alkyl derivatives films swelling. AB - The process of diffusion of sodium hyaluronate and its hydrophobically modified alkyl derivatives in water was studied by measuring the kinetics of solid polymer film swelling. The following fundamental thermodynamic parameters of the swelling process were calculated: the apparent diffusion coefficient of swelling by water D(s) determined at three temperatures (25, 37, and 45 degrees C), the activation enthalpy of diffusion connected with the swelling DeltaH(D,s) and the activation enthalpy of the swelling process DeltaH(s). The thermodynamic activity of the solvent for the given sodium hyaluronate and its four alkyl derivatives has been expressed through the internal quantity RA(delta,s), that is expansion work of polymer coil accomplished by the action of the internal pressure. PMID- 17397040 TI - Tribological investigation of novel HDPE-HAp-Al2O3 hybrid biocomposites against steel under dry and simulated body fluid condition. AB - Among various biocompatible polymers, polyethylene based materials have received wider attention because of its excellent stability in body fluid, inertness, and easy formability. Attempts have been made to improve their physical properties (modulus/strength) to enable them to be used as load bearing hard tissue replacement applications. Among such attempts, high density polyethylene (HDPE) hydroxyapatite (HAp) composite (HAPEX), has already been developed for total hip replacement (THR) acetabular cup and low load bearing bone tissue replacement. In the present work, alumina has been added as a partial replacement of HAp phase to improve the mechanical and tribological properties of the HAPEX composite. In an attempt to assess the suitability of the developed composite in THR application, the tribological properties against steel counterbody under both in air and simulated body fluid (SBF), have been investigated and efforts have been made to understand the wear mechanisms. The fretting wear study indicates the possibility of achieving extremely low COF (Coefficient of Friction approximately 0.09) as well as higher wear resistance (order of 10(-6) mm(3)/N m) with the newly developed composites in SBF. A low wear depth of approximately 4.6-5.3 microm is recorded, irrespective of fretting environment. The implication of the work is that optimal and combined addition of bioactive and bioinert ceramic filler to HDPE can provide a good opportunity to obtain hybrid biocomposites with better combination of physical properties (modulus, hardness) as well as low friction and high wear resistance. PMID- 17397041 TI - Maternal anxiety: course and antecedents during the early postpartum period. AB - The early course and antecedents of postpartum anxiety are unknown. This study sought to determine the course and antecedents of maternal anxiety during the first month postpartum and to develop a model to predict 1-month anxiety using information obtainable before perinatal hospital discharge. Two hundred and ninety-six mothers were screened before discharge with the State (SS) and Trait (TS) Scales of the State Trait Anxiety Inventory (STAI). Demographic characteristics were assessed by questionnaire and medical record review, and psychiatric history, measures of perinatal stress, and resilient factors were determined by focused questions and formal instruments. At 1-month postpartum, the SS was repeated. Scores on the SS were significantly higher at 1 month than immediately postpartum (35.30+/-0.68 versus 33.38+/-0.60, mean+/-standard error, P=.004), but only 58.6% of mothers with high pre-discharge anxiety had high anxiety at 1 month. One-month anxiety correlated with pre-discharge SS and TS scores, a history of psychiatric problems including depressed mood, medical and negative social life events, lack of pregnancy planning and prenatal class attendance, perceived peripartum stress, and duration of postpartum hospital stay. Inverse correlations were observed with education, household income, and resiliency factors. In multivariate modeling, anxiety trait, education, history >or=2 years of depression, and perception of peripartum stress accounted for 50% of the variance in the 1-month SS score. Maternal anxiety increases during the first postpartum month. Women with high trait anxiety, low education, a history of depressed mood, and a perception of high peripartum stress are at risk for experiencing anxiety at this time. PMID- 17397042 TI - Predictive strength of Jonckheere's test for trend: an application to genotypic scores in HIV infection. AB - A problem arising in studies on the human immunodeficiency virus (HIV) infection relate to one-sided tests with ordered alternatives as opposed to the more classical two-sided tests. Patients not having a resistance mutation may have a better virologic response to treatment than patients with a single mutation. In turn, those with a single mutation may have a better response to treatment than those patients having two mutations, and so on. In the presence of a continuous outcome, Jonckheere's test for ordered alternatives is well adapted to this situation. Such an analysis does not provide any measure of prediction or explained variation which can complement these results. A measure of strength of effect would be helpful in quantifying the degree of association between the genotypic score (number of mutations) and some continuous virological response. We suggest a simple measure of 'goodness of split' for Jonckheere's test for trend. Interestingly, the measure can be related to the non-parametric measure of association known as gamma. The variance formula for the measure studied here can be seen to differ from the known variance estimate of the gamma measure, and simulations show it to be more accurate. Expectation and variance under H(0) of the measure are provided and a large simulation study is presented. Methods are applied to a recent clinical data set involving HIV-1 infected patients where the number of resistance mutations are investigated as potential predictors of the amount of HIV-1 RNA reduction at week 4. PMID- 17397043 TI - Spatial memory and hippocampal volume in humans with unilateral vestibular deafferentation. AB - Patients with acquired chronic bilateral vestibular loss were recently found to have a significant impairment in spatial memory and navigation when tested with a virtual Morris water task. These deficits were associated with selective and bilateral atrophy of the hippocampus, which suggests that spatial memory and navigation also rely on vestibular input. In the present study 16 patients with unilateral vestibular deafferentation due to acoustic neurinoma were examined 5- to 13-yrs post-surgery. Volumetry of the hippocampus was performed in patients and age- and sex-matched healthy controls by manually tracing the structure and by an evaluator-independent voxel-based morphometry. Spatial memory and navigation were assessed with a virtual Morris water task. No significant deficits in spatial memory and navigation could be demonstrated in the patients with left vestibular failure, whereas patients with right vestibular loss showed a tendency to perform worse on the respective tests. Impairment was significant only for one computed measure (heading error). The subtle deficiencies with right vestibular loss are compatible with the recently described dominance of the right labyrinth and the vestibular cortex in the right hemisphere. Volumetry did not reveal any atrophy of the hippocampus in either patient group. PMID- 17397044 TI - Maternal experience inhibits the production of immature neurons in the hippocampus during the postpartum period through elevations in adrenal steroids. AB - Motherhood is accompanied by alterations in numerous nonreproductive behaviors, including learning and memory, as well as anxiety and stress regulation. These functions have been linked to adult neurogenesis in the hippocampus, but the effect of maternal experience on this brain region has not been completely explored. To determine whether the production of new hippocampal granule cells is altered during the postpartum period, we examined the number of proliferating cells and their progeny in the dentate gyrus of primiparous female rats at various time points during the postpartum period while they were caring for their offspring, as well as after weaning. Additionally, we investigated whether cell proliferation in the postpartum female is affected by the presence of offspring and nursing-induced increases in glucocorticoids. Analysis of the number of BrdU labeled cells revealed that cell proliferation in the dentate gyrus was suppressed in lactating postpartum females until the time of weaning. This effect was temporary; a difference was detectable at 1 week after BrdU-labeling, when the majority of cells expressed a marker of immature and mature granule neurons (TuJ1) but not at 2 weeks, when most cells expressed a marker of mature neurons (NeuN). The decrease in cell proliferation was dependent on elevated basal glucocorticoid levels associated with lactation; removal of nursing pups reduced basal corticosterone levels and prevented the decrease in the number of BrdU labeled cells. Moreover, preventing increased basal corticosterone levels by means of adrenalectomy and low-dose corticosterone replacement eliminated the reduction in cell proliferation. These findings indicate that offspring interactions inhibit adult neurogenesis through changes in adrenal steroids, and further suggest a potential mechanism for alterations in hippocampal function during the postpartum period. PMID- 17397045 TI - Distribution and abundance of patas monkeys (Erythrocebus patas) in Laikipia, Kenya, 1979-2004. AB - Patas monkeys may be especially vulnerable to local extinction because they live in relatively small, female-philopatric groups at low densities and are strongly polygynous. We assessed a patas monkey population in Kenya's 9,700 km(2) Laikipia District over 25 years, using data collected in 1979-1981 and 1992-2004. The data were based on intensive observations of three study groups, "on the ground" counts, and surveys of Laikipia residents. In 1979-1981, a minimum of 415 patas monkeys lived in 14-15 groups. By 2000, the best estimate suggested 310-445 patas monkeys living in 13-17 groups over a greater surveyed area, suggesting that patas monkeys in Laikipia may have undergone a slight decline in numbers over time. Their distribution, however, was similar over time. The relative stability of this population has likely been the result of beneficial co-existence with large-scale cattle ranching. Outside Laikipia, substantial habitat alteration from rising human populations has coincided with the near disappearance of patas monkeys where they were previously more numerous. The small population in Laikipia, probably the largest remaining in Kenya, may therefore be critical to the continued existence of patas monkeys in that country and may be dependent on maintenance of large-scale ranches. Such land use provides patas monkeys with water and broad expanses of Acacia drepanolobium woodlands, the habitat to which patas are restricted in Laikipia. PMID- 17397046 TI - Ameliorative effects of a neuroprotective agent, T-817MA, on place learning deficits induced by continuous infusion of amyloid-beta peptide (1-40) in rats. AB - Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive decline due to neuronal loss and neural network dysfunction. It has been postulated that progressive neuronal loss in AD is consequence of the neurotoxic properties of the amyloid-beta peptide (Abeta). In the present study, we investigated the effect of T-817MA (1-{3-[2-(1-benzothiophen-5-yl)ethoxy] propyl}-3-azetidinol maleate), a newly synthesized neurotrophic compound, on place learning deficits in rats with hippocampal damages. To induce granule cell loss in the dentate gyrus (DG) of the hippocampus, Abeta (1-40) was continuously infused (300 pmol/day) into the cerebral ventricle using a mini-osmotic pump for 5 weeks. Three weeks after the Abeta infusion, the rats were tested in a place learning task, which required them to alternatively visit two diametrically opposed areas in an open field to obtain intracranial self-stimulation reward. The results indicated that the Abeta-infused rats without treatment of T-817MA displayed learning impairment in the task; their performance level was significantly inferior to that of the vehicle rats. Treatment of T-817MA (8.4 mg/kg/day, p.o.) significantly improved the task performance of the Abeta-infused rats. Furthermore, T-817MA prevented granule cell loss due to Abeta-infusion, which was correlated to task performance of the rats. However, other cognitive enhancer, an acetylcholinesterase inhibitor, had no such effects. The results demonstrated that T-817MA ameliorated learning deficits induced by Abeta infusion, which might be attributed to neuroprotection in the hippocampus. PMID- 17397047 TI - The novel p.L1649Q mutation in the SCN1A epilepsy gene is associated with familial hemiplegic migraine: genetic and functional studies. Mutation in brief #957. Online. AB - Familial hemiplegic migraine (FHM) is a severe subtype of migraine with hemiparesis during attacks. We scanned 10 families with FHM without mutations in the CACNA1A (FHM1) and ATP1A2 (FHM2) genes. We identified the novel p.L1649Q mutation (c.4946T>A) in Na(v)1.1 sodium channel gene SCN1A (FHM3) in a North American kindred with FHM without associated ataxia or epilepsy. Functional analysis of the mutation, introduced in the highly homologous human SCN5A, revealed markedly slowed inactivation and a two-fold faster recovery from fast inactivation predicting enhanced neuronal excitation. Our findings establish the role of neuronal Na(v)1.1 sodium channels in FHM and reinforce the involvement of ion channel dysfunction in the pathogenesis of this episodic brain disorder. PMID- 17397048 TI - Novel SLC4A11 mutations in patients with recessive congenital hereditary endothelial dystrophy (CHED2). Mutation in brief #958. Online. AB - Autosomal recessive congenital hereditary endothelial dystrophy (CHED2) is a severe and rare corneal disorder that presents at birth or shortly thereafter, characterized by corneal opacification and nystagmus. Recently the gene for CHED2 was identified and seven different mutations in the SLC4A11 gene were reported. Here, we report seven novel mutations and two previously identified mutations in families from India and the United Kingdom with recessive CHED. The novel changes include two nonsense (p.Trp240X; p.Gln800X) three missense (p.Glu143Lys; p.Cys386Arg; p.Arg755Trp) and two splice site mutations (c.2240+1G>A; c.2437 1G>A). Interestingly, the c.2398C>T (p.Gln800X) and c.2437-1G>A identified in two affected siblings represent the first compound heterozygous mutations in the SLC4A11 gene. PMID- 17397049 TI - Mutational analysis of CACNA1G in idiopathic generalized epilepsy. Mutation in brief #962. Online. AB - Recent studies have strongly implicated low voltage-activated/T-type calcium channels (T-channels) in the etiology of epilepsy. Here, we report the results of a mutational analysis of the CACNA1G gene, encoding the T-channel Ca(V)3.1/(1G) subunit, using a cohort of 123 mostly Japanese and Hispanic patients with idiopathic generalized epilepsies (IGE) and 360 healthy control individuals. We found 13 variants, including five which involved amino acid substitutions. One variant, c.1709C>T (Ala570Val), is present in a sporadic case of juvenile myoclonic epilepsy (JME) with early childhood absence and astatic seizures, but was not found in control samples. Another variant, c.3265G>T (Ala1089Ser), was observed in three family members affected with JME, and also in one control individual. Two JME patients and three control individuals harbored a third variant, c.2968G>A (Asp980Asn). Although not statistically significant, slightly faster inactivation decay rates were observed in some mutant channels. Our collective findings flag CACNA1G as a potential susceptibility locus for IGE subsyndromes that warrants closer investigation. PMID- 17397050 TI - Mutational analysis of the HGSNAT gene in Italian patients with mucopolysaccharidosis IIIC (Sanfilippo C syndrome). Mutation in brief #959. Online. AB - Mucopolysaccharidosis (MPS) describes any inherited lysosomal storage disorder resulting from an inability to catabolize glycosaminoglycans. MPS III (or Sanfilippo syndrome) is an autosomal recessive disease caused by a failure to degrade heparan sulphate. There are four subtypes of MPS III, each categorized by a deficiency in a specific enzyme involved in the heparan sulphate degradation pathway. The genes mutated in three of these (MPS IIIA, MPS IIIB, and MPS IIID) have been cloned for some time. However, only very recently has the gene for MPS IIIC (heparin acetyl CoA: alpha-glucosaminide N-acetyltransferase, or HGSNAT) been identified. Its product (previously termed transmembrane protein 76, or TMEM76) has little sequence similarity to other proteins of known function, although it is well conserved among all species. In this study, a group of MPS IIIC patients, who are mainly of Italian origin, have been clinically characterized. Furthermore, mutational analysis of the HGSNAT gene in these patients resulted in the identification of nine alleles, of which eight are novel. Three splice-site mutations, three frameshift deletions resulting in premature stop codons, one nonsense mutation, and two missense mutations were identified. The latter are of particular interest as they are located in regions which are predicted to be of functional significance. This research will aid in determining the molecular basis of HGSNAT protein function, and the mechanisms underlying MPS IIIC. PMID- 17397051 TI - Spectrum of MKS1 and MKS3 mutations in Meckel syndrome: a genotype-phenotype correlation. Mutation in brief #960. Online. AB - Meckel syndrome (MKS) is a rare autosomal recessive lethal condition characterized by central nervous system malformations (typically occipital meningoencephalocele), postaxial polydactyly, multicystic kidney dysplasia, and ductal proliferation in the portal area of the liver. MKS is genetically heterogeneous and three loci have been mapped respectively on 17q23 (MKS1), 11q13 (MKS2), and 8q24 (MKS3). Very recently, two genes have been identified: MKS1/FLJ20345 on 17q in Finnish kindreds, carrying the same intronic deletion, c.1408-35_c.1408-7del29, and MKS3/TMEM67 on 8q in families from Pakistan and Oman. Here we report the genotyping of the MKS1 and MKS3 genes in a large, multiethnic cohort of 120 independent cases of MKS. Our first results indicate that the MKS1 and MKS3 genes are each responsible for about 7% of MKS cases with various mutations in different populations. A strong phenotype-genotype correlation, depending on the mutated gene, was observed regarding the type of central nervous system malformation, the frequency of polydactyly, bone dysplasia, and situs inversus. The MKS1 c.1408-35_1408-7del29 intronic mutation was identified in three cases from French or English origin and dated back to 162 generations (approx. 4050 years) ago. We also identified a common MKS3 splice site mutation, c.1575+1G>A, in five Pakistani sibships of three unrelated families of Mirpuri origin, with an estimated age-of-mutation of 5 generations (125 years). PMID- 17397052 TI - Molecular analysis and characterization of nine novel CTSK mutations in twelve patients affected by pycnodysostosis. Mutation in brief #961. Online. AB - Molecular characterization of twelve unrelated patients affected by the autosomal recessive osteosclerotic skeletal dysplasia, Pycnodysostosis (cathepsin k deficiency), revealed 11 different genotypes. The mutational profile consisted of 12 different mutations, including nine previously unreported ones, spread throughout the whole gene. One mutation occurred in regions coding predomain, two affected the prodomain and nine others occurred in the mature domain. The novel lesions consisted in six missense mutations c.20T>C (p.L7P), c.494A>G (p.Q165R), c.580G>A (p.G194S), c.746T>C (p.I249T), c.749A>G (p.D250G), c.955G>T (p.G319C), two frameshifts c.60_61dupGA (p.I21RfsX29), c.282dupA (p.S95VfsX9) and a splicing mutation c.890G>A (r.785_890del). The six new missense mutations were examined by western blots of COS-7 cells transfected with mutant CTSK genes. The L7P, occurring within the predicted hydrophobic domain of signal peptide, showed a significantly reduced expression level compared to the wild type control. These findings suggested that the mutation affected targeting and translocation of the nascent lysosomal protein across the endoplasmatic reticulum membrane. The novel amino acid changes were also modeled into the three-dimensional structure that predicted incorrect protein folding for all of them. Molecular characterization of the patients is of particular value for genetic counseling of patients and their families as diagnosis of Pycnodysostosis based on enzyme assay is unpractical and thus not offered routinely. PMID- 17397053 TI - Novel mutations in FRMD7 in X-linked congenital nystagmus. Mutation in brief #963. Online. AB - Congenital nystagmus is an eye movement disorder in which one or both eyes are in constant movement. It can be associated with a number of ocular or neurological diseases, or it can be inherited in an autosomal or X-linked fashion. The latter form is called idiopathic or motor nystagmus (CIN). Loci on the X chromosome (NYS1) and on 6p12 (NYS2), 7p11.2 (NYS3), and 13q31-q33 (NYS4) have been identified for CIN. The molecular characterization of NYS1 has recently been solved by Tarpey et al., who identified mutations in FRMD7, a gene of unclear function. We report five novel mutations in FRMD7 and confirm the role of this gene in the pathogenesis of X-linked congenital nystagmus. PMID- 17397054 TI - Screening for the BRCA1-ins6kbEx13 mutation: potential for misdiagnosis. Mutation in brief #964. Online. AB - Misdiagnosis of a germline mutation associated with an inherited disease syndrome can have serious implications for the clinical management of patients. A false negative diagnosis (mutation missed by genetic screening) limits decision making about intervention strategies within families. More serious is the consequence of a false positive diagnosis (genetic test suggesting a mutation is present when it is not). This could lead to an individual, falsely diagnosed as a mutation carrier, undergoing unnecessary clinical intervention, possibly involving risk reducing surgery. As part of screening 283 ovarian cancer families for BRCA1 mutations, we used two different methods (mutation specific PCR and multiplex ligation-dependent probe amplification) to screen for a known rearrangement mutation L78833.1:g.44369_50449dup (ins6kbEx13). We found false positive and false negative results in several families. We then tested 61 known carriers or non-carriers from an epidemiological study of BRCA1 and BRCA2 mutation carriers (the EMBRACE study). These data highlight the need for caution when interpreting analyses of the ins6kbEx13 mutation and similar mutations, where characterising the exact sequence alteration for a deleterious mutation is not a part of the routine genetic test. PMID- 17397055 TI - Allelic heterogeneity of molecular events in human coagulation factor IX in Asian Indians. Mutation in brief #965. Online. AB - Mutations in Factor IX gene (F9) cause X-linked recessive bleeding disorder hemophilia B. Here, we characterized molecular events in nine North Indian hemophiliac families identifying four missense mutations (three novel), two nonsense mutations, and a deletion. We have also captured the mutational spectrum of this disease in India based on available reports and established their genotype/phenotype relationships. Indian F9 mutations data indicate the absence of an important germline mutagen in the Indian subcontinent over the last century, and are consistent with previously made conclusions that universal, presumably endogenous factors are predominant in the causation of the spontaneous mutations in F9. We also analyzed the distribution of Ala194Thr polymorphism in 1231 Asian Indians and have established that Ala variant is far more frequent and can certainly be exploited for carrier detection, contrary to earlier reports. PMID- 17397056 TI - Real-SPINE: an integrated system of neural networks for real-value prediction of protein structural properties. AB - Proteins can move freely in three-dimensional space. As a result, their structural properties, such as solvent accessible surface area, backbone dihedral angles, and atomic distances, are continuous variables. However, these properties are often arbitrarily divided into a few classes to facilitate prediction by statistical learning techniques. In this work, we establish an integrated system of neural networks (called Real-SPINE) for real-value prediction and apply the method to predict residue-solvent accessibility and backbone psi dihedral angles of proteins based on information derived from sequences only. Real-SPINE is trained with a large data set of 2640 protein chains, sequence profiles generated from multiple sequence alignment, representative amino-acid properties, a slow learning rate, overfitting protection, and predicted secondary structures. The method optimizes more than 200,000 weights and yields a 10-fold cross-validated Pearson's correlation coefficient (PCC) of 0.74 between predicted and actual solvent accessible surface areas and 0.62 between predicted and actual psi angles. In particular, 90% of 2640 proteins have a PCC value greater than 0.6 between predicted and actual solvent-accessible surface areas. The results of Real-SPINE can be compared with the best reported correlation coefficients of 0.64-0.67 for solvent-accessible surface areas and 0.47 for psi angles. The real SPINE server, executable programs, and datasets are freely available on http://sparks.informatics.iupui.edu. PMID- 17397057 TI - Identification of near-native structures by clustering protein docking conformations. AB - Most state-of-the-art protein-protein docking algorithms use the Fast Fourier Transform (FFT) technique to sample the six-dimensional translational and rotational space. Scoring functions including shape complementarity, electrostatics, and desolvation are usually exploited in ranking the docking conformations. While these rigid-body docking methods provide good performance in bound docking, using unbound structures as input frequently leads to a high number of false positive hits. For the purpose of better selecting correct docking conformations, we structurally cluster the docking decoys generated by four widely-used FFT-based protein-protein docking methods. In all cases, the selection based on cluster size outperforms the ranking based on the inherent scoring function. If we cluster decoys from different servers together, only marginal improvement is obtained in comparison with clustering decoys from the best individual server. A collection of multiple decoy sets of comparable quality will be the key to improve the clustering result from meta-docking servers. PMID- 17397058 TI - The N- and C-termini of the human Nogo molecules are intrinsically unstructured: bioinformatics, CD, NMR characterization, and functional implications. AB - RTN4 or Nogo proteins are composed of three alternative splice forms, namely 1192 residue Nogo-A, 373-residue Nogo-B, and 199-residue Nogo-C. Nogo proteins have received intense attentions because they have been implicated in a variety of critical cellular processes including CNS neuronal regeneration, vascular remodeling, apoptosis, interaction with beta-amyloid protein converting enzyme, and generation/maintenance of the tubular network of the endoplasmic reticulum (ER). Despite their significantly-different N-terminal lengths, they share a conserved C-terminal reticulon-homology domain consisting of two transmembrane fragments, a 66-residue extracellular loop Nogo-66 and a 38-residue C-tail carrying ER retention motif. Nogo-A owns the largest N-terminus with 1016 residues while the Nogo-B has an N-terminus almost identical to the first 200 residues of Nogo-A. So far, except for our previous determination of the Nogo-66 solution structure, no structural characterization of the other Nogo regions has been reported. In the present study, we initiated a systematically investigation of structural properties of Nogo molecules by a combined use of bioinformatics, CD, and NMR spectroscopy. The results led to two striking findings: (1) in agreement with bioinformatics prediction, the N- and C-termini of Nogo-B were experimentally demonstrated to be intrinsically unstructured by CD, two dimensional 1H 15N NMR HSQC, hydrogen exchange, and 15N heteronuclear NOE characterization. (2) Further studies showed that the 1016-residue N-terminus of Nogo-A was again highly disordered. Therefore, it appears that being intrinsically-unstructured allows Nogo molecules to serve as double-faceted functional players, with one set of functions involved in cellular signaling processes essential for CNS neuronal regeneration, vascular remodeling, apoptosis and so forth and with another in generating/maintaining membrane-related structures. We propose that this mechanism may represent a general strategy to place the formation/maintenance of membrane-related structures under the direct regulation of the cellular signaling. PMID- 17397059 TI - A critical review of the pharmacology of the plant extract of Pygeum africanum in the treatment of LUTS. AB - Despite an unremitting increase in the number of patients presenting symptoms of benign prostate hyperplasia (BPH), the viable treatment options remain relatively limited when compared to other disorders of aging. This has spurred an interest in so-called alternative medicines, many of which continue to be used in spite of the more recent emergence of rationally targeted therapies. Nonetheless, in the case of plant extracts, the vast majority of these have not been subjected to the same rigorous pre-clinical pharmacological testing and large-scale clinical trials now required by health authorities. Furthermore, demonstration of their clinical efficacy in BPH has been hindered by trials of limited duration with a high placebo response. Beginning with a preliminary demonstration of in vitro inhibition of growth factor-mediated fibroblast proliferation with Pygeum africanum extract, a detailed series of in vitro and in vivo studies on prostate growth and bladder function were undertaken. These studies, reviewed herein, have permitted the identification of putative molecular targets of Pygeum africanum extract affecting both growth factor-mediated prostate growth as well as specific parameters of bladder function. These results, corroborated in part by short-term clinical efficacy, set the stage for a large-scale clinical trial to investigate the efficacy of Pygeum africanum extract in the treatment of lower urinary tract symptoms. PMID- 17397060 TI - Flexible relaxation of rigid-body docking solutions. AB - Molecular Dynamics (MD) simulations have been performed on a set of rigid-body docking poses, carried out over 25 protein-protein complexes. The results show that fully flexible relaxation increases the fraction of native contacts (NC) by up to 70% for certain docking poses. The largest increase in the fraction of NC is observed for docking poses where anchor residues are able to sample their bound conformation. For each MD simulation, structural snap-shots were clustered and the centre of each cluster used as the MD-relaxed docking pose. A comparison between two energy-based scoring schemes, the first calculated for the MD-relaxed poses, the second for energy minimized poses, shows that the former are better in ranking complexes with large hydrophobic interfaces. Furthermore, complexes with large interfaces are generally ranked well, regardless of the type of relaxation method chosen, whereas complexes with small hydrophobic interfaces remain difficult to rank. In general, the results indicate that current force-fields are able to correctly describe direct intermolecular interactions between receptor and ligand molecules. However, these force-fields still fail in cases where protein-protein complexes are stabilized by subtle energy contributions. PMID- 17397061 TI - Randomized, double-blind, sham-controlled evaluation of the effect of functional continuous magnetic stimulation in patients with urgency incontinence. AB - AIMS: To evaluate the effect of functional continuous magnetic stimulation (FCMS) on urgency incontinence in randomized, sham-controlled manner. METHODS: Thirty nine patients with urgency incontinence, 16 males and 23 females (aged 66.0 +/- 16.5 years), who were refractory to pelvic floor muscle training (PFMT), were randomly assigned either to the treatment schedule performing 10-week active treatment, followed by 4-week non-treatment interval and then by 10-week sham treatment (A-S, n = 20) or to that performing the sham treatment first followed by 10-week active treatment (S-A, n = 19). RESULTS: At 10 weeks, the number of leaks/week, the total score of the International Consultation on Incontinence Questionnaire: Short Form (ICIQ-SF), and maximum cystometric capacity (MCC) were significantly improved as compared with the initial levels (P < 0.001, P < 0.001, and P = 0.003, respectively) in the former group, but not in the latter group. Four (20.0%) patients were cured in the A-S group, while no patient was cured in the S-A group. At the end of the A-S schedule (24 weeks of study), the effect of the active treatment was still maintained at a significantly improved level, as compared with the initial level. At the end of the S-A schedule, the number of leaks/week was significantly improved as compared with the initial level and with its 10-week level (P < 0.001 and P = 0.049, respectively), as well as ICIQ-SF total score (P = 0.001 and P = 0.006, respectively). MCC significantly increased from its initial level (P = 0.030). CONCLUSION: Magnetic stimulation was effective on urgency incontinence in comparison to sham stimulation in this small patient group. PMID- 17397062 TI - Characterization of interfacial solvent in protein complexes and contribution of wet spots to the interface description. AB - Water networks in protein interfaces can complement direct interactions contributing significantly to molecular recognition, function, and stability of protein association. Thus, water can be seen as an extension or addition of protein structural features, which may add plenty of information to protein interfacial definition. However, solvent is frequently neglected in protein interaction studies. Analysis of the interfacial information contained in the PDB is essential to achieve more accurate descriptions of protein interfaces. With this aim, we have used the SCOWLP database (http://www.scowlp.org) and applied computational geometry methods to extract and analyze interfacial information of a high-resolution nonredundant dataset of 176 protein complexes containing obligate and transient interfaces. We have identified all interfacial residues and characterized them in terms of temperature factors, secondary structure, residue composition, and pairing preferences to understand their contribution to the interface description. We have paid special attention to water-bridged residues; focusing on those that interact only mediated by a water molecule called wet spots. Our results show that 40.1% of the interfacial residues are interacting through water and that wet spots represent a 14.5% of the total, emphasizing the importance of the inclusion of solvent in protein interaction studies, and the contribution of wet spots to interfacial description. Wet spots present similar characteristics to residues binding buried water molecules in the core or cavities of proteins; being preferably located in nonregular secondary structures and establishing hydrogen bonds by their main-chains. We observe that obligate and transient interfaces present a comparable amount of solvent. Moreover, the role of solvent in both complex types differs according to the different nature of their interfaces. The information obtained in our studies will assist in the process of accomplishing more accurate descriptions of protein interfaces and may be helpful to improve comparison of protein family interfaces, to facilitate rational ligand design, and to guide protein docking. PMID- 17397063 TI - Tracking cell proliferation using the far red fluorescent dye SNARF-1. AB - BACKGROUND: The [(3)H]thymidine incorporation assay and staining of living cells with fluorescent dyes like carboxyfluorescein diacetate, succinimidyl ester (CFSE) have evolved as valuable methods for studying T cell responses. To assess proliferation of cells already labeled by FITC, CFSE, GFP, or other "green" molecules or to simultaneously track two otherwise indistinguishable cell populations in mixed cell cultures, it would be desirable to have a dye with distinct fluorescent properties for this application. METHODS: We analyzed the dilution of the far red fluorescent dye SNARF-1 in proliferating cells by flow cytometric analysis. The results were compared with the CFSE dilution technique as well as the [(3)H]thymidine incorporation assay. RESULTS: Staining of primary human lymphocytes revealed that SNARF-1 labeling was equivalent to CFSE for estimating proportions of proliferating cells in stimulated cell cultures and yielded results comparable to [(3)H]thymidine incorporation. We showed that SNARF 1 offers the possibility to simultaneously analyze the proliferation of phenotypically indistinguishable subsets of hematopoietic cells and can also be used to track uniformly proliferating, non hematopoietic cells like HEK293. CONCLUSIONS: In summary, we have demonstrated that labeling of cells with SNARF-1 allows for estimating cell proliferation of cells of hematopoietic and non hematopoietic origin. PMID- 17397064 TI - Luminex technology for anti-HLA antibody screening: evaluation of performance and of impact on laboratory routine. AB - The recent introduction of new technologies such as Luminex has provided alternative methods to the Complement Dependent Cytotoxicity (CDC) test for HLA specific antibody detection. In this study we compared the results obtained with CDC to those obtained using a Luminex method with the aim of evaluating the impact of this new technology on antibody screening policies in our transplant setting.A total of 1,421 sera, acquired from patients on the waiting list for a kidney transplant or following transplantation, were tested by both methodologies. CDC was performed using a whole lymphocyte population comprising a panel of 52 cells. The percentage panel reactive antibodies (PRA) and antibody specificity were evaluated using Lambda Scan Analysis software. For the Luminex method sera screening and identification of antibody specificity were carried out using the LABScreen Mixed and LABScreen PRA respectively. The overall concordance between the results obtained using the CDC and the Luminex methods was 85%. HLA antibody specificity was confirmed in 96% of the sera which tested positive using the Luminex system and serum positivity corresponded with a previous sensitisation event in these individuals. Using the Luminex method 18% of patients on the waiting list were considered and managed as sensitised as compared to 7% when testing with CDC alone. The Luminex method was able to detect a number of antibody specificities significantly more frequently than the CDC method and in addition the CDC method failed to detect some of the antibody specificities detected by the Luminex system. Based on this comparison study we have incorporated the Luminex methodology into our screening strategy. PMID- 17397065 TI - Flow cytometric CD34+ stem cell enumeration: lessons from nine years' external quality assessment within the Benelux countries. AB - BACKGROUND: A biannual external quality assurance (EQA) scheme for flow cytometric CD34+ haematopoietic stem cell enumeration has been operational in the Benelux countries since 1996. In an evaluation of the results of 16 send-outs, we studied the effects of the methods used on assay outcome and whether or not this exercise was effective in reducing between-laboratory variation. METHODS: Data were analyzed using robust multivariate regression. This approach is relatively insensitive to outliers and is used to assess the effect of methodological aspects of CD34+ cell counting on the bias and variability. RESULTS: Five variables were associated with significant bias of absolute CD34+ cell counts: (i) unique laboratory number (ULN), (ii) gating strategy; (iii) CD34 mAb fluorochrome; (iv) type of flow cytometer, and (v) method of sample preparation. In addition, ULN and platform methodology (i.e., single vs. dual) contributed significantly to the variability of this assay. Overall, the variability in results of CD34+ cell enumeration has declined with time; in particular, after a practical workshop in which participants were trained to use the "single platform ISHAGE protocol." CONCLUSIONS: Between-laboratory variation in CD34+ cell enumeration can be reduced by standardization of methodologies between centres. Our approach, i.e., EQA with targeted training and feedback in response to reported results, has been successful in reducing the variability of CD34+ cell enumeration between participants. PMID- 17397066 TI - Expression of annual cycles in preen wax composition in red knots: constraints on the changing phenotype. AB - Birds living in seasonal environments change physiology and behavior in correspondence to temporally changing environmental supplies, demands and opportunities. We recently reported that the chemical composition of uropygial gland secretions of sandpipers (Scolopacidae, order Charadriformes) changes during the breeding season from mixtures of monoesters to diesters, which fulfill specific functions related to incubation. A proper temporal match between the expression of diester preen waxes and incubation requires a flexible organization of the trait. Here we analyze the possible degrees of flexibility with reference to the functionality of better-understood molt and body mass cycles of free living and captive red knots (Calidris canutus). The relative flexibility of seasonal cycles in preen wax composition was examined by two experimental perturbations: (1) giving birds restricted access to food and (2) monitoring them long-term under a constant photoperiodic regime. We found that wax type cannot change instantaneously, but that changing the type of wax is under similar organizational time constraints as the replacement of feathers. Just as molt and mass cycles, the seasonal rhythm of diester secretion appeared to be under endogenous control: most birds placed in a constant photoperiod still maintained seasonally changing preen waxes. Diester preen wax secretion was synchronized with the peak in body mass in spring, but became less well expressed under constant photoperiodic conditions and when food availability was limited. PMID- 17397067 TI - Drosophila melanogaster and Eucypris virens giant spermatozoa as visualized by cell inclusion in microgels. AB - A new technique, based on live Sperm Inclusion in Microgels (SIM), allows quick and easy analysis of giant spermatozoa under bright field or fluorescence microscopy. The technique has been assayed on Drosophila melanogaster (Diptera) and Eucypris virens (Ostracoda) spermatozoa and based on the inclusion of freshly obtained male gametes in low melting agarose microgels at 37 degrees C to prevent cell damage. Gametes spread onto pretreated slides are dehydrated and directly observed under phase contrast microscopy or stained with specific fluorochromes for DNA or proteins. Results show that the morphology of whole sperm is highly preserved allowing identification of sperm characteristics difficult to visualize after using standard fixation procedures. In Drosophila melanogaster, SIM allows the simultaneous visualization of the complete flagellum plus a clear delineation of the DNA. It also allows observation of morphological changes in the perforatorium as sperm elongation takes place. In E. virens, SIM slides visualized under phase contrast and fluorescence microscopy show three main morphological regions on the entire spermatozoa. Spermatozoa of this species are auto-fluorescent under wavelength excitation from 387 to 562 nm. In spite of this, clear localization of the DNA molecule at the posterior 1/3 part of the whole spermatozoa can also be achieved after single DAPI staining or in combination with Mercuri-diBrom-Fluorescein, a fluorochrome for protein targeting. PMID- 17397068 TI - Architectural gear ratio and muscle fiber strain homogeneity in segmented musculature. AB - In the segmented axial musculature of fishes and amphibians, the patterns of muscle fiber shortening depend on both the orientation of muscle fibers relative to the long axis of the body as well as the distance of fibers from the neutral axis of bending (vertebral column). In this study we use the relatively simple architecture of salamander hypaxial muscles to explore the separate and combined effects of these morphological features on muscle fiber strains during swimming. In Siren lacertina the external oblique (EO) muscle has more obliquely oriented muscle fibers and is located further from the neutral axis of bending than the internal oblique (IO) muscle. To examine the effect of muscle fiber angle on strain patterns during swimming, we used sonomicrometry to quantify architectural gear ratio (AGR=longitudinal strain/fiber strain) in these two hypaxial muscles. By comparing the muscle fiber strains and shortening velocities of the EO and IO during swimming, we test whether variation in mediolateral position of the muscle layers is counteracted by their differences in AGR. We find that despite substantial differences in mediolateral position, the EO and IO undergo similar fiber strains and shortening velocities for a given amount of axial bending. Our results show that variation in muscle fiber angle acts to counteract differences in mediolateral position, thereby minimizing variation in muscle fiber strain and shortening velocity during swimming. These results highlight the significance of both muscle architecture and muscle moment arms in determining the fiber strains required for a given movement. PMID- 17397069 TI - Induction of increase in intracellular calcium concentration of embryonic cells and acceleration of morphogenetic cell movements during amphibian gastrulation by a 50-Hz magnetic field. AB - The influence of an alternating electromagnetic field (EMF) on early development of amphibian embryos was examined. When the embryos developed under the influence of a low-frequency EMF (50 Hz, 5-30 mT), the rate of early development was accelerated. The effect of EMF was exerted preferentially at the gastrula stage, and the period of gastrulation was shortened. Histological observations showed that EMF promoted morphogenetic cell movements during the gastrulation. The concentration of intracellular free Ca2+ ([Ca2+]i) in the embryonic cells under the influence of EMF was analyzed using Fura-2, an indicator of the intracellular concentration of calcium ions. The influence of EMF on [Ca2+]i was analyzed in embryonic cells isolated from blastula, gastrula, and neurula, EMF increased a [Ca2+]i particularly in the cells isolated from gastrula. Our results suggest that EMF specifically increased the [Ca2+]i of gastrula cells, thereby, accelerating the rate of morphogenetic cell movements during gastrulation. PMID- 17397070 TI - Sequence, expression and evolutionary relationships of carbamoyl phosphate synthetase I in the toad Xenopus laevis. AB - The sequence of carbamoyl phosphate synthetase I (CPSase I) cDNA and expression of the enzyme in liver of the toad Xenopus laevis are reported. CPSase I mRNA increases 6-fold when toads are exposed to high salinity for extended periods of time. The deduced 1,494-amino acid sequence of the CPSase I is homologous to other CPSases and reveals a domain structure and conserved amino acids common to other CPSases. A serine residue (S287) is present where there is a cysteine residue required for glutamine-dependent activity in CPSase Types III and II (Type I CPSases utilize only ammonia as nitrogen-donating substrate). A sequence of DNA 964 bases upstream from the ATG start codon for the CPSase I gene is also reported. Phylogenetic analysis for 30 CPSase isoforms, including X. laevis CPSase I, across a wide spectrum of phyla is reported and discussed. The results are consistent with the views that eukaryotic CPSase II as a multifunctional complex evolved from prokaryotic CPSase II and that CPSase I in terrestrial vertebrates and CPSase III in fishes arose from eukaryotic CPSase II by independent events after the divergence of plants in eukaryotic evolution. PMID- 17397071 TI - Absorption of tetraethylammonium (TEA+) by perfused lobster intestine. AB - The organic cation, tetraethylammonium (TEA(+)), is actively secreted by mammalian nephrons and crustacean urinary bladders by similar processes in both animal groups. These mechanisms consist of a basolateral Organic Cation Transporter (OCT family) that employs the transmembrane electrical potential as a driving force for organic cation uptake from the blood and a brush border secondary active transport process that exchanges luminal protons for TEA(+). The present study examined the nature of (14)C-TEA(+) transport across the perfused intestinal epithelium of the American lobster, Homarus americanus, to ascertain whether the gut complemented the kidneys in the clearance of these organic metabolites from the blood. Unidirectional mucosa to serosa (M to S) (14)C-TEA(+) fluxes in anterior and posterior intestine were hyperbolic functions of luminal [TEA(+)] and significantly (P<0.01) exceeded the respective serosa to mucosa (S to M) fluxes. Luminal quinine (1 mM) significantly (P<0.05) inhibited M to S flux of the organic cation, while serosal addition of the drug had no effect on S to M transfer of TEA(+). Reducing serosal pH from 7.20 to 6.02 significantly (P<0.01) stimulated M to S transfer of 0.1 mM (14)C-TEA(+), but significantly (P<0.05) lowered S to M transfer of the metabolite. Addition of 2.0 mM unlabelled serosal TEA(+) trans-stimulated the M to S flux of 0.1 mM (14)C-TEA and doubled the transfer rate of the organic cation from lumen to blood compared to its transport in the absence of TEA(+) in the bath. Results suggest that this organic cation is absorbed across lobster intestine by the combination of a brush border OCT-1-like transporter coupled with a basolateral H(+)/TEA(+) exchanger. A working model is presented for intestinal organic cation absorption in crustaceans and compared to the secretory transport model for this class of metabolites previously reported for crustacean and mammalian kidneys. PMID- 17397072 TI - Overcoming thermodynamic and kinetic limitations of aldolase-catalyzed reactions by applying multienzymatic dynamic kinetic asymmetric transformations. PMID- 17397073 TI - Domino N-H/C-H bond activation: palladium-catalyzed synthesis of annulated heterocycles using dichloro(hetero)arenes. PMID- 17397074 TI - Estimation of the kinetic acidity from substrate conformation--stereochemical course of the deprotonation of cyclohexenyl carbamates. PMID- 17397075 TI - Is eka-mercury (element 112) a group 12 metal? PMID- 17397076 TI - Convergent chemical synthesis and crystal structure of a 203 amino acid "covalent dimer" HIV-1 protease enzyme molecule. PMID- 17397077 TI - A simple approach for the selective and sensitive colorimetric detection of anionic surfactants in water. PMID- 17397078 TI - Chemiluminescent energy-transfer cassettes based on fluorescein and nile red. PMID- 17397079 TI - A thermally adjustable multicolor photochromic hydrogel. PMID- 17397080 TI - Improving solid-supported catalyst productivity by using simplified packed-bed microreactors. PMID- 17397081 TI - Layer-by-layer constructed macroporous architectures. PMID- 17397082 TI - Space- and time-resolved visualization of acid catalysis in ZSM-5 crystals by fluorescence microscopy. PMID- 17397083 TI - Anisotropically phase-segregated Pd-Co-Pd sulfide nanoparticles formed by fusing two Co-Pd sulfide nanoparticles. PMID- 17397084 TI - Combating Alzheimer's disease with multifunctional molecules designed for metal passivation. PMID- 17397085 TI - A 9-borylated acridinyl radical. PMID- 17397086 TI - Domino ring-opening/recyclization reactions of doubly activated cyclopropanes as a strategy for the synthesis of furoquinoline derivatives. PMID- 17397087 TI - Copper-catalyzed synthesis of N-sulfonyl-1,2,3-triazoles: controlling selectivity. PMID- 17397088 TI - Accelerating reactions with microreactors at elevated temperatures and pressures: profiling aminocarbonylation reactions. PMID- 17397089 TI - syn-selective and enantioselective direct cross-aldol reactions between aldehydes catalyzed by an axially chiral amino sulfonamide. PMID- 17397090 TI - Recombinant fusion proteins TAT-Mu, Mu and Mu-Mu mediate efficient non-viral gene delivery. AB - BACKGROUND: The inherent ability of certain peptides or proteins of viral, prokaryotic and eukaryotic origin to bind DNA was used to generate novel peptide based DNA delivery protocols. We have developed a recombinant approach to make fusion proteins with motifs for DNA-binding ability, Mu and membrane transduction domains, TAT, and tested them for their DNA-binding, uptake and transfection efficiencies. In one of the constructs, the recombinant plasmid was designed to encode the Mu moiety of sequence MRRAHHRRRRASHRRMRGG in-frame with TAT of sequence YGRKKRRQRRR to generate TAT-Mu, while the other two constructs, Mu and Mu-Mu, harbor a single copy or two copies of the Mu moiety. METHODS: Recombinant his-tag fusion proteins TAT-Mu, Mu and Mu-Mu were purified by overexpression of plasmid constructs using cobalt-based affinity resins. The peptides were characterized for their size and interaction with DNA, complexed with plasmid pCMVbeta-gal, and shown to transfect MCF-7, COS and CHOK-1 cells efficiently. RESULTS: Recombinant fusion proteins TAT-Mu, Mu and Mu-Mu were cloned and overexpressed in BL21(DE3)pLysS with greater than 95% purity. The molecular weight of TAT-Mu was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) to be 11.34 kDa while those of Mu and Mu-Mu were 7.78 and 9.83 kDa, respectively. Live uptake analysis of TAT-Mu, Mu and Mu-Mu as DP (DNA+peptide) or DPL (DNA+peptide+lipid) complexes into MCF-7 cells, followed by immunostaining and laser scanning confocal microscopy, demonstrated that the complexes are internalized very efficiently and localized in the nucleus. DNA:peptide complexes (DP) transfect MCF-7, COS and CHOK-1 cells. The addition of cationic liposomes enhances the uptake of the ternary complexes (DPL) further and also brings about 3-7-fold enhancement in reporter gene expression compared to DP alone. CONCLUSIONS: Recombinant proteins that are heterologous fusions, having DNA-binding domains and nuclear localization epitopes, generated in this study have considerable potential to facilitate DNA delivery and enhance transfection. The domains in these fusion proteins would be promising in the development of non-viral gene delivery vectors particularly in cells that do not divide. PMID- 17397091 TI - The administration of naked plasmid DNA into the liver induces antitumor innate immunity in a murine liver metastasis model. AB - BACKGROUND: Gene therapy is a promising strategy against advanced cancer; however, the safety of viral vectors and the effectiveness of non-viral vectors have not yet been established. Recently, a hydrodynamics-based procedure was reported to be an effective and safe method to deliver and transduce DNA into the liver. Herein, we propose a strategy for liver metastasis by a hydrodynamics based procedure to deliver naked non-coding plasmid DNA (pDNA) into the liver as an immunocompetent organ. METHODS AND RESULTS: Mice received a rapid intravenous (i.v.) injection of naked pDNA in a large volume of saline (0.1 ml/g body weight). The single administration of a naked non-coding pDNA by the hydrodynamics-based procedure before tumor cell inoculation strongly suppressed liver metastasis formation. However, the usual i.v. injection (200 microl/body) of the same dose of naked pDNA could not suppress liver metastasis formation. Following the methylation of CpG sequences within the pDNA using CpG methylase, injection of the methylated pDNA by the hydrodynamics-based procedure could not suppress liver metastasis formation. Gadolinium chloride pretreatment did not interfere with this antitumor effect, but anti-asialo GM1 antiserum treatment did. These findings indicated that natural killer (NK) cells, not Kupffer cells, were involved in this antitumor effect. The NK cytotoxic activities of liver mononuclear cells were strongly enhanced after receiving a naked pDNA by the hydrodynamics-based procedure. CONCLUSIONS: These observations suggest that unmethylated CpG motifs in pDNA stimulated immune cells, resulting in the activation of NK cells in the liver to suppress liver metastases in a murine model. PMID- 17397092 TI - Development of strategies for conditional RNA interference. AB - BACKGROUND: RNA interference (RNAi) represents a powerful tool with which to undertake sequence-dependent suppression of gene expression. Synthesized double stranded RNA (dsRNA) or dsRNA generated endogenously from plasmid or viral vectors can be used for RNAi. For the latter, polymerase III promoters which drive ubiquitous expression in all tissues have typically been adopted. Given that dsRNA molecules must contain few 5' and 3' over-hanging bases to maintain potency, employing polymerase II promoters to drive tissue-specific expression of RNAi may be problematic due to potential inclusion of nucleotides 5' and 3' of siRNA sequences. METHODS: To circumvent this, polymerase II promoters in combination with cis-acting hammerhead ribozymes and short-hairpin RNA sequences have been explored as a means to generate potent dsRNA molecules in tissues defined by the promoter in use. RESULTS: The novel constructs evaluated in this study produced functional siRNA which suppressed the enhanced green fluorescent protein (eGFP) both in vitro and in vivo (in mice). Additionally, the constructs did not appear to elicit a significant type-1 interferon response compared to traditional H1-transcribed shRNA. CONCLUSIONS: Given the potential 'off-target' effects of dsRNAs, it would be preferable in many cases to limit expression of dsRNA to the tissue of interest and moreover would significantly augment the resolution of RNAi technologies. Notably, the system under evaluation in this study could readily be adapted to achieve this objective. PMID- 17397093 TI - Care-giver advice as a preventive measure for drinking during pregnancy: zeros, categorical outcome responses, and endogeneity. AB - We conduct an empirical investigation of the impact of prenatal care-giver advice on alcohol consumption by pregnant women. In the design of the model and estimator, we pay particular attention to three aspects of the data. First, a large proportion of pregnant women do not drink at all. To accommodate this aspect of the sample we base the essential formulation of the model on the modified version of the two-part approach of Duan et al. (Journal of Business and Economic Statistics 1983; 1: 115-126.) suggested by Mullahy (Journal of Health Economics 1998; 17: 247-281.). Second, in the survey that we analyze (the 1988 National Maternal and Infant Health Survey - NMIHS), respondents were only required to report their consumption up to a specified range of values (e.g. 1-2 drinks per week, 2-5 drinks per week, and so on). For this reason, the model is cast in the grouped regression framework of Stewart (Review of Economic Studies 1983; 50: 141-149.). Third, the binary physician advice variable is likely to be endogenous and the econometric specification explicitly accounts for this possibility. To summarize the results, we find that failing to account for endogeneity leads to the counterintuitive conclusion that advice has a positive and statistically significant influence on drinking during pregnancy. When the model is extended to allow for potential endogeneity, we find that advice has a negative and statistically significant impact. PMID- 17397096 TI - Effects of chronic drinking on verb generation: an event related potential study. AB - In alcohol dependent individuals, abnormalities in brain functioning have been revealed using event-related potential (ERP) methods. In the present study, we investigated whether in non-alcohol dependent drinkers functioning of the brain is also compromised as a function of recent and lifetime drinking history (LDH). An ERP verb generation task consisting of two conditions (generating verbs describing the use of visually presented nouns versus reading nouns aloud) was used; subtracting ERPs in the latter condition from those in the former should reveal the sequence of brain processes involved in verb generation. Four groups were included, consisting of individuals drinking either lightly, moderately, heavily, or excessively (overall mean age 46.6 years). Participants were sober at the time of testing. Although the excessive group had the highest per cent retrieval errors, there was no continuous relationship between this score and amount of alcohol consumption. However, number of glasses per week affected differential ERPs associated with verb generation both at short (120-220 ms, mid frontal sites) and at longer latencies (from 700 ms on),left-temporal and right frontal electrode sites (T7, F6). It is concluded that moderate, heavy, and excessive drinkers, compared to light drinkers, show abnormal brain potentials associated with verb generation over frontal and temporal areas. Moderate to excessive drinking alters some but not all brain processes involved in verb generation. In particular the frontal and temporal brain areas appear to be vulnerable for the effects of chronic lifetime drinking. PMID- 17397097 TI - High and low dosage oxcarbazepine versus naltrexone for the prevention of relapse in alcohol-dependent patients. AB - INTRODUCTION: Oxcarbazepine (OXC) reduces high-voltage-activated calcium currents, thus reducing glutamatergic transmission at corticostriatal synapses. This effect on NMDA glutamatergic transmission may play a role against the increased glutamatergic transmission determined by alcohol withdrawal. To investigate the efficacy and safety of OXC in relapse prevention we compared OXC at different dosages with Naltrexone (NAL) in a 90 days randomised open-label trial. Craving and psychiatric symptoms improvements were the secondary endpoints. METHODS: Eighty-four detoxified alcohol dependent subjects currently meeting clinical criteria for alcohol dependence were randomised into three groups: 27 patients received 50 mg of naltrexone, 29 received 1500-1800 mg of oxcarbazepine (OXC high), 28 patients 600-900 mg of oxcarbazepine (OXC low). Craving (VAS; OCDS) and withdrawal (AWRS) rating scales were applied; psychiatric symptoms were evaluated through the SCL-90-R. RESULTS: A significantly larger number of subjects remained alcohol free in the OXC high group (58.6%) with respect to both the OXC low (42.8%) and the NAL groups (40.7%). Comparing the OCDS total scores at the end of the treatment, the improvement was significantly greater for the NAL group with respect to the OXC low group. The reduction of the Hostility-Aggression subscore of the SCL-90-R was significantly greater in the OXC high group than that of the other groups. Dual diagnosis patients had a better outcome when treated with OXC high. DISCUSSION: OXC at a dosage of 1500 1800 mg/day might be beneficial in terms of alcohol relapse prevention. The low dosage formulation did not show the same trend, but it still remain in the same range as NAL. The mechanism involved in the efficacy of oxcarbazepine in relapse prevention could be less related to craving and more connected to the treatment of the comorbid psychiatric symptomatology and the alcohol protracted withdrawal syndrome. PMID- 17397098 TI - Exploring the clinical and social determinants of prescribing anticholinergic medication for Chinese patients with schizophrenia. AB - AIMS: To-date few studies have investigated prescription patterns of anticholinergic medication (ACM) in Chinese patients with schizophrenia in general and outpatients in particular. This study examined the frequency and socio-demographic and clinical correlates of ACM in Hong Kong (HK) and Beijing (BJ), China. METHODS: Five hundred and five clinically stable outpatients with schizophrenia were randomly selected and interviewed in HK and BJ using standardized assessment instruments. Basic socio-demographic and clinical data and psychotropic drug prescriptions were collected at the time of the diagnostic interview. RESULTS: ACM was found in 47.7% of (n = 241) the whole sample and in 54.1 and 41.2% of the HK and BJ samples, respectively. ACM was associated with more frequent use of antipsychotic (AP) polypharmacy, typical AP drugs, and a combination of depot and oral AP, less use of clozapine and atypical and oral AP, a lack of health insurance, higher doses of APs, severity of extrapyramidal side effects (EPS), a higher number of APs prescribed, and study sites. In multiple logistic regression analysis, higher doses of APs, less frequent use of an atypical AP, and study sites all remained significantly associated with ACM. CONCLUSION: Although the ethnic and clinical characteristics of the two samples were nearly identical, there was a wide variation in the frequency of ACM prescriptions between HK and BJ suggesting that socio-cultural and economical factors as well as traditions of psychiatric practice all played a role in determining the use of ACM. Prescribing ACM in neither site was in line with current recommendation. PMID- 17397099 TI - Effects of smoking marijuana on focal attention and brain blood flow. AB - Using an attention task to control cognitive state, we previously found that smoking marijuana changes regional cerebral blood flow (rCBF). The present study measured rCBF during tasks requiring attention to left and right ears in different conditions. Twelve occasional marijuana users (mean age 23.5 years) were imaged with PET using [15O]water after smoking marijuana or placebo cigarettes as they performed a reaction time (RT) baseline task, and a dichotic listening task with attend-right- and attend-left-ear instructions. Smoking marijuana, but not placebo, resulted in increased normalized rCBF in orbital frontal cortex, anterior cingulate, temporal pole, insula, and cerebellum. RCBF was reduced in visual and auditory cortices. These changes occurred in all three tasks and replicated our earlier studies. They appear to reflect the direct effects of marijuana on the brain. Smoking marijuana lowered rCBF in auditory cortices compared to placebo but did not alter the normal pattern of attention related rCBF asymmetry (i.e., greater rCBF in the temporal lobe contralateral to the direction of attention) that was also observed after placebo. These data indicate that marijuana has dramatic direct effects on rCBF, but causes relatively little change in the normal pattern of task-related rCBF on this auditory focused attention task. PMID- 17397100 TI - Cardiovascular effects of high dose venlafaxine XL in patients with major depressive disorder. AB - OBJECTIVE: To assess cardiovascular safety profile of high dose Venlafaxine XL in patients with major depression. METHOD: Effects of high dose venlafaxine (mean 346.15 mg;) on the cardiovascular system in 37 patients with major depressive disorder were evaluated: BP, ECG (PR, QT, QRSD and QTc intervals) and heart rate. RESULTS: 12.5% of patients developed hypertension after starting treatment with venlafaxine. There was an association between heart rate and the dose of venlafaxine although not statistically significant. There was no association between dose of venlafaxine and PR, QT, QRSD and QTc intervals. One patient on 300 mg who was hypertensive and on other medications that may prolong QTc, had mildly prolonged QTc. However this was not clinically significant. CONCLUSION: This study of subjects on high dose venlafaxine (mean 346.15 mg; range 225-525 mg) did not demonstrate any clinical or statistically significant effects on electrocardiogram (ECG) parameters including PR, QT, QRSD and QTc interval. PMID- 17397101 TI - Serotonin reuptake inhibitors in pregnancy and the risk of major malformations: a systematic review. AB - OBJECTIVE: To review studies conducted to establish the risk of major congenital malformations in women exposed to serotonin reuptake inhibitors (SRIs) during the first trimester of pregnancy. METHODS: A literature search [corrected] was conducted within PsycINFO [corrected] EMBASE, MEDLINE and Cochrane databases from 1966 to October 2006, to identify studies assessing the risk of major malformations in infants whose mother was taking SRIs (SSRIs and SNRIs) during the first trimester of pregnancy. RESULTS: Fifteen studies were selected for the analysis: seven adopted a prospective cohort design and seven a retrospective design, of these one was a case-control study. DATA SYNTHESIS AND CONCLUSIONS: The reviewed studies suggest that exposure to fluoxetine, sertraline, citalopram and venlafaxine in early pregnancy is not associated with an increased risk of major congenital malformations. For paroxetine, recent data call for caution in prescribing such a drug in early pregnancy. For the other SRIs, the risk remains substantially undetermined, as data are so far scanty. Given this background, large prospective cohort studies are urgently needed to better assess the risk/benefit ratio of SRIs-treatment during pregnancy. PMID- 17397102 TI - Hypoxia-targeted over-expression of carboxylesterase as a means of increasing tumour sensitivity to irinotecan (CPT-11). AB - The induced expression of carboxylesterase (CE) enzymes, which convert the prodrug irinotecan (CPT-11) into its active cytotoxic metabolite SN-38, constitutes a promising strategy for cancer gene therapy. By incorporating hypoxia-responsive elements (HREs) in conjunction with the transgene, expression can be targeted specifically to hypoxic tissues (such as solid tumours), expressing the hypoxia-inducible factor 1 (HIF-1). We have constructed a recombinant adenoviral vector, AdHRE-rCE, encoding the cDNA for the highly efficient rabbit liver CE (rCE), under the control of a HRE derived from the human phosphoglycerate kinase 1 (PGK-1) gene in conjunction with a minimal SV40 promoter. In vitro, HT1080 fibrosarcoma and SW480 colon carcinoma cells demonstrated an approximately 10-fold hypoxia-dependent induction in CE expression following pre-infection with AdHRE-rCE, which led to a15-30-fold increased sensitivity to CPT-11. Furthermore, in vivo, SW480 tumour xenografts infected with AdHRE-rCE demonstrated a 2-fold decrease in tumour doubling time, when combined with 7 days of CPT-11 treatment, in comparison to mock-infected controls, with rCE expression shown to be limited to hypoxic regions only. As the cytotoxicity of CPT-11 is reduced under hypoxic conditions, over-expression of a highly efficient CE such as rCE under hypoxia control within these hypoxic cells could reverse this effect and, therefore, form the basis for future clinical treatment strategies. PMID- 17397103 TI - Nuclear localisation and pDNA condensation in non-viral gene delivery. AB - BACKGROUND: Non-viral gene delivery vectors are multi-component systems reflecting various functionalities required for effective cell transfection, including DNA condensation, promotion of cell membrane interactions and provision for subcellular targeting through endosomal escape and/or nuclear delivery. Elements mediating these functions will clearly display inter-dependency. In this study we sought to explore the relationship within non-viral vectors of condensation and nuclear localisation. METHODS: Binary, tertiary and quaternary vectors were prepared with combinations of pDNA, DOTAP lipid, the polycation peptide protamine and either SV40 nuclear localisation sequence peptide ('SV40 NLS') or a one amino acid substituted mutant of SV40 NLS ('mutant sequence'). The efficiency of pDNA condensation was determined by gel electrophoresis and quantitative fluorescence spectroscopy. Transfection efficiency was examined in mammalian cells in vitro using standard methods, by electroporation to bypass the plasma membrane barrier and in cells arrested in G0/G1 cell cycle phase to examine the effect of cell division and nuclear membrane disruption. RESULTS: Small NLS peptide sequences, despite possessing a significant proportion of basic amino acids, display minimal pDNA-condensing ability when compared to larger polycations such as protamine. In standard in vitro cell adherent transfection studies the predominant elements affording enhanced gene expression were effective pDNA condensation and lipid enhancement of cell membrane interactions. These features conversely hinder efficient gene expression in cells that have undergone electroporation. The benefit of SV40 NLS was only apparent when used in non-dividing cell populations. CONCLUSIONS: Whilst effective levels of non-viral mediated gene expression generally rely on efficient condensation of pDNA and enhanced interactions with cellular membranes, non-covalently associated NLS within a multi-component non-viral gene vector appears to contribute benefit in sustaining gene expression in non-dividing cells. PMID- 17397104 TI - Novel chitosan derivative nanoparticles enhance the immunogenicity of a DNA vaccine encoding hepatitis B virus core antigen in mice. AB - BACKGROUND: Chitosan has been shown to possess useful properties such as non toxicity, high biocompatibility and non-antigenicity that offer advantages for vaccine delivery systems. In this study, we prepared novel chitosan derivative nanoparticles as DNA vaccine carriers and the potential and mechanism of the DNA nanoparticle complexes in inducing augmented immune responses were explored. METHODS: The pVAX(HBc)DNA-nanoparticle complexes as vaccine delivery systems were studied in several aspects: the protection against DNase I degradation was measured by an in vitro inhibition assay; the sustained expression of the plasmid in vivo was determined by RT-PCR; the elevated uptake efficiency by phagocytes was observed with confocal microscopy; the biocompatibility was evaluated by cytotoxicity and histology assay; the complexes were administrated to C57BL/6 mice and the humoral and cellular immune responses were evaluated by ELISA, IFN gamma production and cytolytic T lymphocyte (CTL)-specific lysis assay. RESULTS: The remaining relative activity of DNase I after inhibition varied from 32.3% to 77.6%. The complexes were observed with higher uptake efficiency by phagocytes than naked DNA. Three types of nanoparticles did not induce significant cytotoxicity at concentrations2) stages for their characterization. Derivatization remains critical to position substructures in a glycan array since product ions carry fragmentation "scars" throughout the MSn tree. Equally as important are the pathway relationships between each stage and the greater yield of fragments with the smaller number of oscillators. Applications were directed to the structural isomers in ovalbumin and IgG, where, in the latter case, several previously unreported glycans were detected. Procedures were supported with bioinformatics tools for assimilating structure from the MSn data sets. PMID- 17397138 TI - Probing structural differences in prion protein isoforms by tyrosine nitration. AB - Two conformational isomers of recombinant hamster prion protein (residues 90-232) have been probed by reaction with two tyrosine nitration reagents, peroxynitrite and tetranitromethane. Two conserved tyrosine residues (tyrosines 149 and 150) are not labeled by either reagent in the normal cellular form of the prion protein. These residues become reactive after the protein has been converted to the beta-oligomeric isoform, which is used as a model of the fibrillar form that causes disease. After conversion, a decrease in reactivity is noted for two other conserved residues, tyrosine 225 and tyrosine 226, whereas little to no effect was observed for other tyrosines. Thus, tyrosine nitration has identified two specific regions of the normal prion protein isoform that undergo a change in chemical environment upon conversion to a structure that is enriched in beta sheet. PMID- 17397139 TI - Denaturation of protein by chlorine dioxide: oxidative modification of tryptophan and tyrosine residues. AB - Oxychlorine compounds, such as hypochlorous acid (HOCl) and chlorine dioxide (ClO2), have potent antimicrobial activity. Although the biochemical mechanism of the antimicrobial activity of HOCl has been extensively investigated, little is known about that of ClO2. Using bovine serum albumin and glucose-6-phosphate dehydrogenase of Saccharomyces cerevisiae as model proteins, here I demonstrate that the antimicrobial activity of ClO2 is attributable primarily to its protein denaturing activity. By solubility analysis, circular dichroism spectroscopy, differential scanning calorimetry, and measurement of enzymatic activity, I demonstrate that protein is rapidly denatured by ClO2 with a concomitant decrease in the concentration of ClO2 in the reaction mixture. Circular dichroism spectra of the ClO2-treated proteins show a change in ellipticity at 220 nm, indicating a decrease in alpha-helical content. Differential scanning calorimetry shows that transition temperature and endothermic transition enthalpy of heat-induced unfolding decrease in the ClO2-treated protein. The enzymatic activity of glucose 6-phosphate dehydrogenase decreases to 10% within 15 s of treatment with 10 microM ClO2. Elemental analyses show that oxygen, but not chlorine, atoms are incorporated in the ClO2-treated protein, providing direct evidence that protein is oxidized by ClO2. Furthermore, mass spectrometry and nuclear magnetic resonance spectroscopy show that tryptophan residues become N-formylkynurenine and tyrosine residues become 3,4-dihydroxyphenylalanine (DOPA) or 2,4,5 trihydroxyphenylalanine (TOPA) in the ClO2-treated proteins. Taking these results together, I conclude that microbes are inactivated by ClO2 owing to denaturation of constituent proteins critical to their integrity and/or function, and that this denaturation is caused primarily by covalent oxidative modification of their tryptophan and tyrosine residues. PMID- 17397140 TI - A novel substrate mimetic inhibitor of PKB/Akt inhibits prostate cancer tumor growth in mice by blocking the PKB pathway. AB - We describe a novel, potent peptide substrate mimetic inhibitor of protein kinase B (PKB/Akt). The compound selectively kills prostate cancer cells, in which PKB is highly activated, but not normal cells, or cancer cells in which PKB is not activated. The inhibitor induces apoptosis and inhibits the phosphorylation of PKB substrates in prostate cancer cell lines and significantly increases the efficacy of chemotherapy agents to induce prostate cancer cell death, when given in combination. In vivo, the inhibitor exhibits a strong antitumor effect in two prostate cancer mouse models. Moreover, treated animals develop significantly less lung metastases compared to untreated ones, and the effect is accompanied by a significant decrease in blood PSA [prostate-specific antigen] levels in treated animals. This compound and its potential analogues may be developed into novel, potent, and safe anticancer agents, both as stand-alone treatment and in combination with other chemotherapy agents. PMID- 17397141 TI - Interactions of isopenicillin N synthase with cyclopropyl-containing substrate analogues reveal new mechanistic insight. AB - Isopenicillin N synthase (IPNS), a non-heme iron oxidase central to penicillin and cephalosporin biosynthesis, catalyzes an energetically demanding chemical transformation to produce isopenicillin N from the tripeptide delta-(l-alpha aminoadipoyl)-l-cysteinyl-d-valine (ACV). We describe the synthesis of two cyclopropyl-containing tripeptide analogues, delta-(l-alpha-aminoadipoyl)-l cysteinyl-beta-methyl-d-cyclopropylglycine and delta-(l-alpha-aminoadipoyl)-l cysteinyl-d-cyclopropylglycine, designed as probes for the mechanism of IPNS. We have solved the X-ray crystal structures of these substrates in complex with IPNS and propose a revised mechanism for the IPNS-mediated turnover of these compounds. Relative to the previously determined IPNS-Fe(II)-ACV structure, key differences exist in substrate orientation and water occupancy, which allow for an explanation of the differences in reactivity of these substrates. PMID- 17397142 TI - Kinetic characterization of human JNK2alpha2 reaction mechanism using substrate competitive inhibitors. AB - Jun N-terminal kinase (JNK) is a stress activated serine/threonine protein kinase that phosphorylates numerous cellular protein substrates including the transcription factors c-Jun and ATF2. In this study, we defined the kinetic mechanism for the active form of JNK2alpha2. Double reciprocal plots of initial rates versus concentrations of substrate revealed the sequential nature of the JNK2alpha2 catalyzed ATF2 phosphorylation. Dead-end JNK inhibitors were then used to differentiate ordered and random kinetic mechanisms for the reaction. A peptide inhibitor containing the homology JNK docking sequence for substrate recognition, derived from amino acid residues 153-163 of JNK-interacting protein 1 (JIP-1), inhibited JNK activity via competition with ATF2. This peptide functioned as a noncompetitive inhibitor against ATP. In contrast, the anthrapyrazolone compound, SP600125, exhibited competitive inhibition for ATP and noncompetitive inhibition against ATF2. Furthermore, binding of one substrate had no significant effect on the affinity for the other substrate. The data in this study are consistent with a kinetic mechanism for activated JNK2alpha2 in which (1) substrate binding is primarily due to the distal contacts in the JNK2alpha2 docking groove that allow the delivery of the substrate phosphorylation sequence into the catalytic center, (2) there is minimal allosteric communication between the protein-substrate docking site and the ATP binding site in the catalytic center for activated JNK2alpha2, and (3) the reaction proceeds via a random sequential mechanism. PMID- 17397143 TI - Experimental and master equation study of the kinetics of OH + C2H2: temperature dependence of the limiting high pressure and pressure dependent rate coefficients. AB - The kinetics of the reaction OH + C2H2 have been studied using laser flash photolysis at 248 nm to generate OH radicals and laser-induced fluorescence to monitor OH removal. An attempt was made to use the rate coefficients OH (v = 1,2) + C2H2 to determine the limiting high-pressure rate coefficient, k(1a)(infinity), over the temperature range of 195-823 K. This method is usually applicable if the reaction samples the potential energy well of the adduct, HOC2H2, and if intramolecular vibrational relaxation is fast. In the present case, however, the rate coefficients for loss of the vibrationally excited states by reaction with C2H2 also contain a substantial contribution from nonreactive vibrational relaxation, which occurs via a mechanism that does not sample the adduct potential energy well but involves, at least at low temperatures, collisions that access a shallower, longer range van der Waals well. The data were analyzed using a composite mechanism that incorporates both reactive and nonreactive energy transfer mechanisms, which allows the determination of k(1a)(infinity)(T) for OH + C2H2 with satisfactory accuracy over the temperature range 195-823 K. The kinetics of the reaction OH (v = 0) + C2H2 were also studied in He over the range of conditions: 210-373 K and 5-760 Torr. A one-dimensional master equation (ME) analysis of the experimental data provided a further determination of k(1a)(infinity)(T) and also (down) for He. Combining the two sets of results gives a consistent dataset for k(1a)(infinity) and the Arrhenius parameters A1ainfinity = 7.3 x 10(-12) cm(3) molecule(-1) s(-1) and E(1a)(infinity) = 5.3 kJ mol(-1), with (down) = 150(T/300 K) cm(-1). Additional experiments were conducted at room temperature in N(2) and SF(6) by laser flash photolysis with cavity ring down spectroscopy, and ME calculations were then optimized for the pressure falloff in N(2) by varying the average downward energy transfer parameter ((down)). The output from the best fit ME was parametrized using a modified Troe expression to provide rate data for use in atmospheric modeling. PMID- 17397144 TI - Dual cation and anion acceptor molecules. The case of the (eta6 C6H6)(eta6C6F6)Cr0 complex. AB - In this manuscript we report high-level ab initio (RI-MP2(full)/6-31++G**) and DFT (B3LYP/ 6-31++G** and MPWB1K/6-31++G**) calculations on complexes between the bis(arene)chromium complex (eta6-C6H6)(eta6C6F6)Cr(0) (1) and cations/anions. This interesting molecule 1, which is synthetically available, exhibits a dual binding mode to anions and cations, with interaction energies similar to those previously reported for benzene with cations and hexafluorobenzene with anions. In addition, the simultaneous interaction with cations and anions is also studied. PMID- 17397145 TI - Wide compositional and structural diversity in the system Tl/Bi/P/Q (Q=S, Se) and observation of vicinal P-Tl J coupling in the solid state. AB - The compounds alpha-TlBiP2Se6 (I), beta-TlBiP2Se6 (II), TlBiP2S6 (III), Tl3Bi3(PS4)4 (IV), TlBiP2S7 (V), and Tl3Bi(PS4)2 (VI) were synthesized, and the structures of I-V were determined by single-crystal X-ray diffraction analysis. The structure of I features infinite chains. Those of compounds II, III, and V are layered. The structure of IV features a three-dimensional framework. Tl4Bi2(PS4)2(P2S6) (VII) was also prepared for comparison to the title compounds. The band gaps of each compound are 1.23, 1.27, 1.81, 1.88, 2.06, 1.98, and 1.97 eV for I-VII, respectively. Compounds I, III, IV, and VI melt congruently at 544, 595, 495, and 563 degrees C, respectively, and compounds II, V, and VII melt incongruently at 544, 509, and 600 degrees C, respectively. Solid-state 31P NMR spectroscopy of the reported compounds demonstrates chemical shifts and chemical shift anisotropies in line with related chalcophosphate materials. Evidence for two-bond P-Tl J coupling was observed in 31P NMR spectra (J=481-1781 Hz), and to the best of our knowledge, this is the first example of two-bond P-Tl J coupling and the first example of P-Tl coupling in the solid state. It was possible to assign chemical shifts of inequivalent 31P atoms from the same [PxQy]z- anion type based on different modes of metal ion coordination to the chalcogen. These assignments provide information about the vicinal metal ion contribution to the 31P chemical shift. PMID- 17397146 TI - Critical role of water content in the formation and reactivity of uranium, neptunium, and plutonium iodates under hydrothermal conditions: implications for the oxidative dissolution of spent nuclear fuel. AB - The reactions of 237NpO2 with excess iodate under acidic hydrothermal conditions result in the isolation of the neptunium(IV), neptunium(V), and neptunium(VI) iodates, Np(IO3)4, Np(IO3)4.nH2O.nHIO3, NpO2(IO3), NpO2(IO3)2(H2O), and NpO2(IO3)2.H2O, depending on both the pH and the amount of water present in the reactions. Reactions with less water and lower pH favor reduced products. Although the initial redox processes involved in the reactions between 237NpO2 or 242PuO2 and iodate are similar, the low solubility of Pu(IO3)4 dominates product formation in plutonium iodate reactions to a much greater extent than does Np(IO3)4 in the neptunium iodate system. UO2 reacts with iodate under these conditions to yield uranium(VI) iodates solely. The isotypic structures of the actinide(IV) iodates, An(IO3)4 (An=Np, Pu), are reported and consist of one dimensional chains of dodecahedral An(IV) cations bridged by iodate anions. The structure of Np(IO3)4.nH2O.nHIO3 is constructed from NpO9 tricapped-trigonal prisms that are bridged by iodate into a polar three-dimensional framework structure. Second-harmonic-generation measurements on a polycrystalline sample of the Th analogue of Np(IO3)4.nH2O.nHIO3 reveal a response of approximately 12x that of alpha-SiO2. Single-crystal magnetic susceptibility measurements of Np(IO3)4 show magnetically isolated Np(IV) ions. PMID- 17397147 TI - Dendronized diruthenium compounds via the copper(I)-catalyzed click reaction. AB - Novel Ru2-containing dendritic compounds were prepared from the CuI-catalyzed 1,3 dipolar cycloaddition between Ru2 compounds containing one or two ethynes, Ru2(D(3,5-Cl2Ph)F)4-n(DMBA-4-C2H)n with n=1 and 2, and azidopoly(benzyl ether) dendrons. These new compounds were also characterized with X-ray diffraction and voltammetric techniques. PMID- 17397148 TI - Evidence for the formation of terminal hydrides by protonation of an asymmetric iron hydrogenase active site mimic. AB - Treatment of [Fe2(mu-pdt)(CO)6] [pdt=S(CH2)3S] with dppe (Ph2PCH2CH2PPh2) in refluxing toluene affords the asymmetric complex [Fe2(mu-pdt)(CO)4(dppe)] (1). Protonation of 1 with HBF4-Et2O in CH2Cl2 gives at room temperature the mu hydrido derivative [Fe2(mu-pdt)(CO)4(dppe)(mu-H)](BF4) (2). Monitoring the reaction by 1H, 31P, and 13C NMR at low temperature reveals unambiguously that the process of the protonation of 1 implies terminal hydride intermediates. PMID- 17397149 TI - Catalytic double carbonylation of epoxides to succinic anhydrides: catalyst discovery, reaction scope, and mechanism. AB - The first catalytic method for the efficient conversion of epoxides to succinic anhydrides via one-pot double carbonylation is reported. This reaction occurs in two stages: first, the epoxide is carbonylated to a beta-lactone, and then the beta-lactone is subsequently carbonylated to a succinic anhydride. This reaction is made possible by the bimetallic catalyst [(ClTPP)Al(THF)2]+[Co(CO)4]- (1; ClTPP = meso-tetra(4-chlorophenyl)porphyrinato; THF = tetrahydrofuran), which is highly active and selective for both epoxide and lactone carbonylation, and by the identification of a solvent that facilitates both stages. The catalysis is compatible with substituted epoxides having aliphatic, aromatic, alkene, ether, ester, alcohol, nitrile, and amide functional groups. Disubstituted and enantiomerically pure anhydrides are synthesized from epoxides with excellent retention of stereochemical purity. The mechanism of epoxide double carbonylation with 1 was investigated by in situ IR spectroscopy, which reveals that the two carbonylation stages are sequential and non-overlapping, such that epoxide carbonylation goes to completion before any of the intermediate beta-lactone is consumed. The rates of both epoxide and lactone carbonylation are independent of carbon monoxide pressure and are first-order in the concentration of 1. The stages differ in that the rate of epoxide carbonylation is independent of substrate concentration and first-order in donor solvent, whereas the rate of lactone carbonylation is first-order in lactone and inversely dependent on the concentration of donor solvent. The opposite solvent effects and substrate order for these two stages are rationalized in terms of different resting states and rate-determining steps for each carbonylation reaction. PMID- 17397151 TI - Use of p-toluenesulfonic acid for the controlled grafting of alkoxysilanes onto silanol containing surfaces: preparation of tunable hydrophilic, hydrophobic, and super-hydrophobic silica. AB - The modification of Aerosil 200 has been carried out using methoxysilanes in toluene reflux, with p-toluenesulfonic acid as the catalyst. Both trimethoxyalkyl silanes (methyl, ethyl, propyl, butyl, hexyl, octyl, and octadecyl) and trialkylmethoxy silanes (trimethyl and dimethyloctyl) have been used. The surface has been studied by 29Si NMR, 13C NMR, elemental analysis, thermogravimetry, water contact angle, and BET analysis. When incorporating trimethoxysilanes, a plateau of modification was achieved after 1 h of reflux, while when using trialkylmethoxy silanes, a longer time of about 7 h was required. The average number of molecules incorporated in both cases has been well above those reported by other authors in similar reactions and in much shorter times. Depending on the modification agent and on the experimental conditions, the resulting organosilicas are in seven cases superhydrophobic, in three cases hydrophobic, and in two cases hydrophilic. Two structural origins for superhydrophobicity have been identified in these samples: almost complete disappearance of water accessible surface silanols (smallest methoxysilanes) and shielding of would-be water accessible surface silanols by long aliphatic tails. These features can be very precisely controlled. PMID- 17397150 TI - Targeted drug delivery utilizing protein-like molecular architecture. AB - Nanotechnology-based drug delivery systems (nanoDDSs) have seen recent popularity due to their favorable physical, chemical, and biological properties, and great efforts have been made to target nanoDDSs to specific cellular receptors. CD44/chondroitin sulfate proteoglycan (CSPG) is among the receptors overexpressed in metastatic melanoma, and the sequence to which it binds within the type IV collagen triple-helix has been identified. A triple-helical "peptide-amphiphile" (alpha1(IV)1263-1277 PA), which binds CD44/CSPG, has been constructed and incorporated into liposomes of differing lipid compositions. Liposomes containing distearoyl phosphatidylcholine (DSPC) as the major bilayer component, in combination with distearoyl phosphatidylglycerol (DSPG) and cholesterol, were more stable than analogous liposomes containing dipalmitoyl phosphatidylcholine (DPPC) instead of DSPC. When dilauroyl phosphatidylcholine (DLPC):DSPG:cholesterol liposomes were prepared, monotectic behavior was observed. The presence of the alpha1(IV)1263-1277 PA conferred greater stability to the DPPC liposomal systems and did not affect the stability of the DSPC liposomes. A positive correlation was observed for cellular fluorophore delivery by the alpha1(IV)1263-1277 PA liposomes and CD44/CSPG receptor content in metastatic melanoma and fibroblast cell lines. Conversely, nontargeted liposomes delivered minimal fluorophore to these cells regardless of the CD44/CSPG receptor content. When metastatic melanoma cells and fibroblasts were treated with exogeneous alpha1(IV)1263-1277, prior to incubation with alpha1(IV)1263-1277 PA liposomes, to potentially disrupt receptor/liposome interactions, a dose dependent decrease in the amount of fluorophore delivered was observed. Overall, our results suggest that PA-targeted liposomes can be constructed and rationally fine-tuned for drug delivery applications based on lipid composition. The selectivity of alpha1(IV)1263-1277 PA liposomes for CD44/CSPG-containing cells represents a targeted-nanoDDS with potential for further development and application. PMID- 17397152 TI - Single-wall carbon nanotubes bearing covalently linked phthalocyanines- photoinduced electron transfer. AB - HiPco single-walled carbon nanotubes (SWNTs) have been sidewall-functionalized with phthalocyanine addends following two different approaches: a straightforward Prato reaction with N-octylglycine and a formyl-containing phthalocyanine, and a stepwise approach that involves a former Prato cycloaddition to the double bonds of SWNTs using p-formyl benzoic acid followed by esterification of the derivatized nanotubes with an appropriate phthalocyanine molecule. The two materials obtained by these routes comprise different carbon/Pc-addenda ratios, as evidenced by Raman, TGA, and photophysical studies. The occurrence of electron transfer from photoexcited phthalocyanines to the nanotube framework in these ZnPc-SWNT ensembles is observed in transient absorption experiments, which confirm the absorption of the one-electron oxidized ZnPc cation and the concomitant bleaching of the van Hove singularities typical from SWNTs. Charge separation (i.e., 2.0 x 1010 s(-1)) and charge-recombination (i.e., 1.5 x 106 s( 1)) dynamics reveal a notable stabilization of the radical ion pair product in dimethylformamide. PMID- 17397153 TI - Proton-directed redox control of O-O bond activation by heme hydroperoxidase models. AB - Hangman metalloporphyrin complexes poise an acid-base group over a redox-active metal center and in doing so allow the "pull" effect of the secondary coordination environment of the heme cofactor of hydroperoxidase enzymes to be modeled. Stopped-flow investigations have been performed to decipher the influence of a proton-donor group on O-O bond activation. Low-temperature reactions of tetramesitylporphyrin (TMP) and Hangman iron complexes containing acid (HPX-CO2H) and methyl ester (HPX-CO2Me) functional groups with peroxyacids generate high-valent Fe=O active sites. Reactions of peroxyacids with (TMP)FeIII(OH) and methyl ester Hangman (HPX-CO2Me)FeIII(OH) give both O-O heterolysis and homolysis products, Compound I (Cpd I) and Compound II (Cpd II), respectively. However, only the former is observed when the hanging group is the acid, (HPX-CO2H)FeIII(OH), because odd-electron homolytic O-O bond cleavage is inhibited. This proton-controlled, 2e- (heterolysis) vs 1e- (homolysis) redox specificity sheds light on the exceptional catalytic performance of the Hangman metalloporphyrin complexes and provides tangible benchmarks for using proton coupled multielectron reactions to catalyze O-O bond-breaking and bond-making reactions. PMID- 17397154 TI - Hybrid gadolinium oxide nanoparticles: multimodal contrast agents for in vivo imaging. AB - Luminescent hybrid nanoparticles with a paramagnetic Gd2O3 core were applied as contrast agents for both in vivo fluorescence and magnetic resonance imaging. These hybrid particles were obtained by encapsulating Gd2O3 cores within a polysiloxane shell which carries organic fluorophores and carboxylated PEG covalently tethered to the inorganic network. Longitudinal proton relaxivities of these particles are higher than the positive contrast agents like Gd-DOTA which are commonly used for clinical magnetic resonance imaging. Moreover these particles can be followed up by fluorescence imaging. This study revealed that these particles suited for dual modality imaging freely circulate in the blood vessels without undesirable accumulation in lungs and liver. PMID- 17397155 TI - Nickel-specific response in the transcriptional regulator, Escherichia coli NikR. AB - Studies of the transcriptional repression of the Ni-specific permease encoded by the Pnik operon by Escherichia coli NikR using a LacZ reporter assay establish that the NikR response is specific to nickel in vivo. Toward understanding this metal ion-specific response, X-ray absorption spectroscopy (XAS) analysis of various M-NikR complexes (M = Co(II), Ni(II), Cu(II), Cu(I), and Zn(II)) was used to show that each high-affinity binding site metal adopts a unique structure, with Ni(II) and Cu(II) being the only two metal ions to feature planar four coordinate complexes. The results are consistent with an allosteric mechanism whereby the geometry and ligand selection of the metal present in the high affinity site induce a unique conformation in NikR that subsequently influences DNA binding. The influence of the high-affinity metal on protein structure was examined using hydrogen/deuterium (H/D) exchange detected by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). Each NikR complex gives rise to differing amounts of H/D exchange; Zn(II)- and Co(II)-NikR are most like apo-NikR, while the exchange time course is substantially different for Ni(II) and to a lesser extent for Cu(II). In addition to the high-affinity metal binding site, E. coli NikR has a low-affinity metal-binding site that affects DNA binding affinity. We have characterized this low-affinity site using XAS in heterobimetallic complexes of NikR. When Cu(II) occupies the high-affinity site and Ni(II) occupies the low-affinity site, the Ni K-edge XAS spectra show that the Ni site is composed of six N/O-donors. A similar low-affinity site structure is found for the NikR complex when Co(II) occupies the low-affinity site and Ni(II) occupies the high-affinity site, except that one of the Co(II) ligands is a chloride derived from the buffer. PMID- 17397156 TI - Solid-state NMR study of amyloid nanocrystals and fibrils formed by the peptide GNNQQNY from yeast prion protein Sup35p. AB - Sup35p is a prion protein found in yeast that contains a prion-forming domain characterized by a repetitive sequence rich in Gln, Asn, Tyr, and Gly amino acid residues. The peptide GNNQQNY7-13 is one of the shortest segments of this domain found to form amyloid fibrils, in a fashion similar to the protein itself. Upon dissolution in water, GNNQQNY displays a concentration-dependent polymorphism, forming monoclinic and orthorhombic crystals at low concentrations and amyloid fibrils at higher concentrations. We prepared nanocrystals of both space groups as well as fibril samples that reproducibly contain three (coexisting) structural forms and examined the specimens with magic angle spinning (MAS) solid-state nuclear magnetic resonance. 13C and 15N MAS spectra of both nanocrystals and fibrils reveal narrow resonances indicative of a high level of microscopic sample homogeneity that permitted resonance assignments of all five species. We observed variations in chemical shift among the three dominant forms of the fibrils which were indicated by the presence of three distinct, self-consistent sets of correlated NMR signals. Similarly, the monoclinic and orthorhombic crystals exhibit chemical shifts that differ from one another and from the fibrils. Collectively, the chemical shift data suggest that the peptide assumes five conformations in the crystals and fibrils that differ from one another in subtle but distinct ways. This includes variations in the mobility of the aromatic Tyr ring. The data also suggest that various structures assumed by the peptide may be correlated to the "steric zipper" observed in the monoclinic crystals. PMID- 17397157 TI - Two-photon absorption in tetraphenylporphycenes: are porphycenes better candidates than porphyrins for providing optimal optical properties for two photon photodynamic therapy? AB - Porphycenes are structural isomers of porphyrins that have many unique properties and features. In the present work, the resonant two-photon absorption of 2,7,12,17-tetraphenylporphycene (TPPo) and its palladium(II) complex (PdTPPo) has been investigated. The data obtained are compared to those from the isomeric compound, meso-tetraphenylporphyrin (TPP). Detection of phosphorescence from singlet molecular oxygen, O2(a(1)Delta(g)), produced upon irradiation of these compounds, was used to obtain two-photon excitation spectra and to quantify two photon absorption cross sections, delta. In the spectral region of 750-850 nm, the two-photon absorption cross sections at the band maxima for both TPPo and PdTPPo, delta = 2280 and 1750 GM, respectively, are significantly larger than that for TPP. This difference is attributed to the phenomenon of so-called resonance enhancement; for the porphycenes, the two-photon transition is nearly resonant with a comparatively intense one-photon Q-band transition. The results of quantum mechanical calculations using density functional quadratic response theory are in excellent agreement with the experimental data and, as such, demonstrate that comparatively high-level quantum chemical methods can be used to interpret and predict nonlinear optical properties from such large molecular systems. One important point realized through these experiments and calculations is that one must exercise caution when using qualitative molecular-symmetry derived arguments to predict the expected spectral relationship between allowed one- and two-photon transitions. From a practical perspective, this study establishes that, in comparison to porphyrins and other tetrapyrrolic macrocyclic systems, porphycenes exhibit many desirable attributes for use as sensitizers in two-photon initiated photodynamic therapy. PMID- 17397158 TI - Mapping the orientation of helices in micelle-bound peptides by paramagnetic relaxation waves. AB - Many antimicrobial peptides form alpha-helices when bound to a membrane. In addition, around 80% of residues in membrane-bound proteins are found in alpha helical regions. The orientation and location of such helical peptides and proteins in the membrane are key factors determining their function and activity. Here we present a new solution state NMR method for obtaining the orientation of helical peptides in a membrane-mimetic environment (micelle-bound) without any chemical perturbation of the peptide-micelle system. By monitoring proton longitudinal relaxation rates upon addition of the freely water-soluble and inert paramagnetic probe Gd(DTPA-BMA) to an alpha-helical peptide, a wavelike pattern with a periodicity of 3.6 residues per turn is observed. The tilt and azimuth (rotation) angle of the helix determine the shape of this paramagnetic relaxation wave and can be obtained by least-square fitting of measured relaxation enhancements. Results are presented for the 15-residue antimicrobial peptide CM15 which forms an amphipathic helix almost parallel to the surface of the micelle. Thus, a few fast experiments enable the identification of helical regions and determination of the helix orientation within the micelle without the need for covalent modification, isotopic labeling, or sophisticated equipment. This approach opens a path toward the topology determination of alpha-helical membrane proteins without the need for a complete NOE-based structure determination. PMID- 17397159 TI - A nearly isosteric photosensitive amide-backbone substitution allows enzyme activity switching in ribonuclease s. AB - psi[CS-NH]4-RNase S, a site specific modified version of RNase S obtained by thioxylation (O/S exchange) at the Ala4-Ala5- peptide bond, was used to evaluate the impact of protein backbone photoswitching on bioactivity. psi[CS-NH](4)-RNase S was yielded by recombination of the S-protein and the respective chemically synthesized thioxylated S-peptide derivative. Comparison with RNase S revealed similar thermodynamic stability of the complex and an unperturbed enzymatic activity toward cytidine 2',3'-cyclic monophosphate (cCMP). Reversible photoisomerization with a highly increased cis/trans isomer ratio of the thioxopeptide bond of psi[CS-NH](4)-RNase S in the photostationary state occurred under UV irradiation conditions (254 nm). The slow thermal reisomerization (t(1/2) = 180 s) permitted us to determine the enzymatic activity of cis psi[CS NH](4)-RNase S by measurement of initial rates of cCMP hydrolysis. Despite thermodynamic stability of cis psi[CS-NH](4)-RNase S, its enzymatic activity is completely abolished but recovers after reisomerization. We conclude that the thioxopeptide bond modified polypeptide backbone represents a versatile probe for site-directed photoswitching of proteins. PMID- 17397160 TI - Evidence for the rapid conversion of stephacidin B into the electrophilic monomer avrainvillamide in cell culture. PMID- 17397161 TI - Real-time observation of the photoinduced structural change of bis(2,9-dimethyl 1,10-phenanthroline)copper(I) by femtosecond fluorescence spectroscopy: a realistic potential curve of the Jahn-Teller distortion. AB - In copper(I) complex [Cu(dmphen)(2)]+ (dmphen = 2,9-dimethyl-1,10 phenanthroline), a "flattening" structural change is induced with 1MLCT excitation, which is a prototype of the structural change accompanied with Cu(I)/Cu(II) conversion in copper complexes. Femtosecond and picosecond emission dynamics of this complex were investigated in solution at room temperature with optically allowed S(2) <-- S(0) photoexcitation. Time-resolved emission was measured in the whole visible region, and the lifetimes, intrinsic emission spectra, and radiative lifetimes of the transients were obtained by quantitative analysis. It was concluded that the initially populated S(2) state is relaxed with a time constant of 45 fs to generate the S1 state retaining the perpendicular structure, and the D(2d) --> D(2) structural change (the change of the dihedral angle between the two ligand planes) occurs in the S(1) state with a time constant of 660 fs. The intersystem crossing from the S(1) state to the T(1) state takes place after this structural distortion with a time constant of 7.4 ps. Importantly, the temporal spectral evolution relevant to the structural change clearly exhibited an isoemissive point around 675 nm. This manifests that there exists a shallow potential minimum at the perpendicular geometry on the S1 surface, and the S1 state stays undistorted for a finite period as long as 660 fs before the structural distortion. This situation is not expected for the structural change induced by the ordinary (pseudo-)Jahn-Teller effect, because the distortion should be induced by the spontaneous structural instability at the perpendicular structure. This result sheds new light on the present understanding on the structural change occurring in the metal complexes. PMID- 17397162 TI - Specificity of sites within eight-membered ring zeolite channels for carbonylation of methyls to acetyls. AB - The acid-catalyzed formation of carbon-carbon bonds from C1 precursors via CO insertion into chemisorbed methyl groups occurs selectively within eight-membered ring (8-MR) zeolite channels. This elementary step controls catalytic carbonylation rates of dimethyl ether (DME) to methyl acetate. The number of O-H groups within 8-MR channels was measured by rigorous deconvolution of the infrared bands for O-H groups in cation-exchanged and acid forms of mordenite (M,H-MOR) and ferrierite (H-FER) after adsorption of basic probe molecules of varying size. DME carbonylation rates are proportional to the number of O-H groups within 8-MR channels. Na+ cations selectively replaced protons within 8-MR channels and led to a disproportionate decrease in carbonylation turnover rates (per total H+). These conclusions are consistent with the low or undetectable rates of carbonylation on zeolites without 8-MR channels (H-BEA, H-FAU, H-MFI). Such specificity of methyl reactivity upon confinement within small channels appears to be unprecedented in catalysis by microporous solids, which typically select reactions by size exclusion of bulkier transition states. PMID- 17397163 TI - Lithographic patterning of photoreactive cell-adhesive proteins. PMID- 17397164 TI - DNA metastability and biological regulation: conformational dynamics of metastable omega-DNA bulge loops. AB - Dynamic interchange between DNA conformations, including metastable states, can be of importance to biological function. In this study, we use a combination of spectroscopic and calorimetric techniques to detect and characterize kinetically trapped, metastable states in strand exchange and strand displacement reactions for bulge loop DNA conformations, here referred to as Omega-DNAs. We show that such metastable, Omega-DNA bulge loop states can stably coexist below 50 degrees C, while rearranging irreversibly at elevated temperatures to thermodynamically more stable states. Such dynamic interchange between metastable and globally stable DNA conformational states can be of importance in biological regulatory mechanisms. PMID- 17397165 TI - Hydroxyapatite surface-induced peptide folding. AB - Herein, we describe the design and surface-binding characterization of a de novo designed peptide, JAK1, which undergoes surface-induced folding at the hydroxyapatite (HA)-solution interface. JAK1 is designed to be unstructured in buffered saline solution, yet undergo HA-induced folding that is largely governed by the periodic positioning of gamma-carboxyglutamic acid (Gla) residues within the primary sequence of the peptide. Circular dichroism (CD) spectroscopy and analytical ultracentrifugation indicate that the peptide remains unfolded and monomeric in solution under normal physiological conditions; however, CD spectroscopy indicates that in the presence of hydroxyapatite, the peptide avidly binds to the mineral surface adopting a helical structure. Adsorption isotherms indicate nearly quantitative surface coverage and Kd = 310 nM for the peptide surface binding event. X-ray photoelectron spectroscopy (XPS) coupled with the adsorption isotherm data suggests that JAK1 binds to HA, forming a self-limiting monolayer. This study demonstrates the feasibility of using HA surfaces to trigger the intramolecular folding of designed peptides and represents the initial stages of defining the design rules that allow HA-induced peptide folding. PMID- 17397166 TI - Self-assembled monolayers of terminal alkynes on gold. PMID- 17397167 TI - An aryl to imidoyl palladium migration process involving intramolecular C-H activation. AB - Biologically interesting fluoren-9-one and xanthen-9-one derivatives have been prepared by a novel aryl to imidoyl palladium migration, followed by intramolecular arylation. The fluoren-9-one synthesis appears to involve both a palladium migration mechanism and a C-H activation process proceeding through an unprecedented organopalladium(IV) hydride intermediate. The results from deuterium labeling experiments are consistent with the proposed dual mechanism. PMID- 17397168 TI - Fluorescent ADP sensing in physiological conditions based on cooperative inhibition of a miniature esterase. PMID- 17397169 TI - Chemical modification of monolayer-protected gold nanoparticles using hyperbaric conditions. PMID- 17397171 TI - Palladium-catalyzed synthesis of highly substituted endocyclic enol lactones via a three-component coupling reaction in an ionic liquid. AB - [reaction: see text] A new, efficient ionic liquid based synthetic procedure was developed for the preparation of highly substituted endocyclic enol lactones via the carbonylation coupling reactions of alkynes and 1,3-diketones in ionic liquid. The reactions proceeded in excellent regioselectivity and in reasonably good yields. The catalyst system can be recycled five times with only modest loss of its catalytic activity. PMID- 17397172 TI - Synthesis and stereochemical assignment of brasilibactin A. AB - [structure: see text] Brasilibactin A, a naturally occurring siderophore related to the mycobactins, has been synthesized in six steps. Use of asymmetric titanium mediated aldol reactions allowed the preparation of both diastereomers from a common synthetic intermediate, thus allowing the relative stereochemistry of the natural product to be assigned. Brasilibactin A exhibits no inhibition of histone deacetylases (HDACs) in spite of the N-formyl-N-hydroxy lysine moiety that is expected to affect the activity of these metal-dependent lysine-modifying enzymes. PMID- 17397173 TI - Syntheses of amamistatin fragments and determination of their HDAC and antitumor activity. AB - [reaction: see text] Amamistatins A and B are natural products found to have anti proliferative effects against MCF-7, A549, and MKN45 human tumor cell lines (IC50 0.24-0.56 microM). It was proposed that their activity was due to histone deacetylase (HDAC) inhibition mediated by the N-formyl-N-hydroxy lysine moiety. Amamistatin B fragment analogs were synthesized and screened for biological activity. These compounds were modest HDAC inhibitors and showed antitumor activity against MCF-7 and PC-3 human tumor cells. PMID- 17397174 TI - From fullerene-mixed peroxide to open-cage oxafulleroid C(59)(O)(3)(OH)(2)(OO(t)()Bu)(2) embedded with furan and lactone motifs. AB - [reaction: see text] Removal of one carbon atom from the C60 cage is achieved under mild conditions. The process involves the formation of fullerene-mixed peroxide, subsequent Lewis acid induced cleavage of O-O and C-O bonds, and thermolysis at 75 degrees C. In the proposed mechanism, the carbon atom is deleted as CO and an oxygen atom occupies the vacancy to form a furan ring. Single-crystal X-ray analysis confirmed the results. PMID- 17397175 TI - Short and efficient synthesis of chiral furyl carbinols from carbohydrates. AB - [reaction: see text] Common sugar derivatives can be transformed in a few steps into chiral-substituted furyl carbinols. The mildness of the reaction conditions and the good yields obtained make this procedure an interesting alternative to the conventional processes. PMID- 17397176 TI - Ruthenium-catalyzed tandem cross-metathesis/Wittig olefination: generation of conjugated dienoic esters from terminal olefins. AB - [reaction: see text] In the presence of ruthenium-based olefin metathesis catalysts and triphenylphosphine, alpha,beta-unsaturated aldehydes can be olefinated with diazoacetates. This ruthenium-catalyzed transformation has been employed in tandem with olefin cross-metathesis to convert terminal olefins into 1,3-dienoic esters in a single operation. PMID- 17397177 TI - Asymmetric synthesis of primary amines via the spiroborate-catalyzed borane reduction of oxime ethers. AB - [reaction: see text] The enantioselective borane reduction of O-benzyloxime ethers to primary amines was studied under catalytic conditions using the spiroborate esters 5-10 derived from nonracemic 1,2-amino alcohols and ethylene glycol. Effective catalytic conditions were achieved using only 10% of catalyst 5 derived from diphenylvalinol in dioxane at 0 degrees C resulting in complete conversion to the corresponding primary amine in up to 99% ee. PMID- 17397178 TI - Total synthesis of the putative structure of stemonidine: the definitive proof of misassignment. AB - [structure: see text] The total synthesis of the putative structure of the Stemona alkaloid stemonidine has been completed. The key transformations include a 1,3-dipolar cycloaddition of a chiral nitrone derived from (S)-prolinol and a spirolactonization process involving the generation of the critical stereocenter. The NMR data of the synthetic material do not match those reported for the natural product. It is concluded that the structure assigned to stemonidine is incorrect, and it must be reassigned as stemospironine. PMID- 17397179 TI - Antioxidant and antimelanogenic activities of polyamine conjugates from corn bran and related hydroxycinnamic acids. AB - The antioxidant activity of three major polyamine conjugates, N,N'-dicoumaroyl putrescine (DCP), N-p-coumaroyl-N'-feruloylputrescine (CFP), and N,N'-diferuloyl putrescine (DFP) isolated from corn bran, and their related hydroxycinnamic acids, p-coumaric acid and ferulic acid, were evaluated by three antioxidant in vitro assay systems, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and superoxide and hydroxyl radicals generated by enzymatic and nonenzymatic reactions. Additionally, five phenolic compounds were evaluated for melanogenesis inhibitory activity using mushroom tyrosinase and B16 melanoma cells. Most of the phenolic compounds significantly scavenged DPPH, superoxide, and hydroxyl radicals in a dose-dependent manner. Particularly, DFP showed potent DPPH (IC50 = 38.46 microM) and superoxide (IC50 = 291.62 microM) radical scavenging activities, while DCP exhibited the strongest hydroxyl radical scavenging activity (IC50 = 120.55 microM). CFP also exerted moderate DPPH, superoxide, and hydroxyl radical scavenging activities. Meanwhile, DCP (IC50 = 181.73 microM) showed potent tyrosinase inhibitory activity toward l-tyrosine as the substrate, whereas DFP (IC50 = 733.64 microM) significantly inhibited melanin synthesis in B16 melanoma cells. These current results indicate that these three polyamine conjugates from corn bran may be useful potential sources of natural antioxidants and skin-whitening agents. PMID- 17397180 TI - Carotenoid accumulation in Japanese apricot (Prunus mume Siebold & Zucc.): molecular analysis of carotenogenic gene expression and ethylene regulation. AB - To elucidate the regulatory mechanisms of carotenogenesis in Japanese apricot (Prunus mume Siebold & Zucc.), the relationships between carotenoid accumulation and the expression of the carotenogenic genes, phytoene synthase (PmPSY-1), phytoene desaturase (PmPDS), zeta-carotene desaturase (PmZDS), lycopene beta cyclase (PmLCYb), lycopene epsilon-cyclase (PmLCYe), beta-carotene hydroxylase (PmHYb), and zeaxanthin epoxidase (PmZEP), were analyzed in two cultivars with different ripening traits, 'Orihime' and 'Nanko.' In 'Orihime' fruits, large amounts of carotenoids accumulated on the tree, concomitant with the induction of PmPSY-1 and the downstream carotenogenic genes PmLCYb, PmHYb, and PmZEP. In 'Nanko' fruits, carotenoids accumulated mainly after harvest, correlating with an appreciable induction of PmPSY-1 expression, but the downstream genes were not notably induced, which may explain the lower total carotenoid content in 'Nanko' than in 'Orihime.' In both cultivars, a decrease in PmLCYe expression and increased or constant PmLCYb expression could cause the metabolic shift from beta,epsilon-carotenoid synthesis to beta,beta-carotenoid synthesis that occurs as ripening approaches. Next, the effects of ethylene on the expression of PmPSY 1 and carotenoid accumulation were investigated in 'Nanko' fruits treated with propylene or 1-methylcyclopropene (1-MCP). Propylene treatment induced both ethylene production and carotenoid accumulation. PmPSY-1 was constitutively expressed, but propylene treatment accelerated its induction. 1-MCP treatment caused a slight inhibition of carotenoid accumulation along with the repression, although not complete, of PmPSY-1. Collectively, although PmPSY-1 expression was not exclusively regulated by ethylene, both the notable induction of PmPSY-1 accelerated by ethylene and the subsequent induction of the downstream carotenogenic genes, especially PmLCYb, could be necessary for the massive carotenoid accumulation that occurs during ripening. Furthermore, the switch from PmLCYe expression to PmLCYb expression could cause beta,beta-carotenoid accumulation in both Japanese apricot cultivars. PMID- 17397181 TI - High-throughput method for the quantitation of total folate in whole blood using LC-MS/MS. AB - A high-throughput liquid chromatography tandem mass spectrometry (HT LC-MS/MS) method for red blood cell (RBC) folate analysis was developed from a previously described manual (M) LC-MS/MS method. The HT LC-MS/MS method used 96-well plates in which RBC folates were hydrolyzed with concentrated HCl in the presence of the [13C6]pABA internal standard (IS). The pH of the hydrolysate was adjusted to 5.0 before cleanup using 96-well plate OASIS HLB SPE cartridges. The analyte and IS were eluted with ethyl acetate/hexane (95:5, v/v) and methylated with methanol and trimethylsilyldiazomethane. The methylated analyte and IS were quantified with LC-MS/MS as previously described. The HT LC-MS/MS method was validated by determining the recovery of six different folate vitamers, which were quantitatively recovered (84-105% with CV<9.0%). RBC folate concentrations in whole blood samples correlated between HT and M LC-MS/MS methods (r=0.922, p<0.0001 for n=43 samples) and between the HT LC-MS/MS method and a chemiluminescence assay (r=0.664, p<0.001 for n=325 samples). Comparison of the results between HT LC-MS/MS and chemiluminescence methods with Bland-Altman difference plots and by ROC curve analysis indicates that the chemiluminescence assay underreports RBC folate concentrations. The HT LC-MS/MS method allows for high-throughput sample preparation for the analysis of RBC folate. PMID- 17397182 TI - Comparison of two dietary folate intake instruments and their validation by RBC folate. AB - An optimal folate nutritional status is important in minimizing developmental and degenerative disease. Therefore, constant monitoring of folate intake and of biomarkers of folate nutritional status is essential. The objective of this research was to compare two folate intake instruments and validate each one against RBC folate measured by a high-throughput liquid chromatography tandem mass spectrometry (HT LC-MS/MS) method described in the companion paper (Owens, J. E.; Holstege, D. M.; Clifford, A. J. J. Agric. Food Chem. 2007, 55, 3292 3297). A food frequency questionnaire (FFQ) and a folate-targeted semiquantitative Block dietary folate equivalents (DFE) screener were compared and individually validated against an HT LC-MS/MS method. RBC folate was 1178 +/- 259 nmol/L (mean +/- SD) in a population of 337 normal adult subjects. Folate intakes were 556 +/- 265 microg/day by the FFQ and 524 +/- 276 microg/day by the DFE screener. Folate intakes by the DFE screener were approximately 34 microg less than by the FFQ (paired t test, p<0.01), but the intake instruments were highly correlated for total folate intake (r=0.608, p<0.01). Correlations between instruments and RBC folate were low (r<0.35) but strong (p<0.01). ROC curve analysis indicates that the measurement of RBC folate by the HT LC-MS/MS method is a better predictive tool than are intake instruments for the evaluation of marginal folate status. PMID- 17397183 TI - Interaction of resveratrol and its trimethyl and triacetyl derivatives with biomembrane models studied by differential scanning calorimetry. AB - The interaction of resveratrol (trans-3,5,4'-trihydroxystilbene) and two of its derivatives (3,5,4'-tri-O-methylresveratrol and 3,5,4'-tri-O triacetylresveratrol) with biomembrane models, represented by dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles (MLV), has been studied by differential scanning calorimetry (DSC). The analysis of MLV prepared in the presence of increasing molar fraction of such compounds has been carried out to reveal their maximum interaction with biomembrane models. The results from these studies have been compared with kinetic experiments results, in order to detect the entity and rate of compound absorption by the biomembrane models. The findings indicate that the compounds affected the thermotropic properties of DMPC MLV by suppressing the pretransition peak and broadening the DMPC main phase transition calorimetric peak and shifting it to lower temperatures. The order of effectiveness found was resveratrol > trimethylresveratrol > triacetylresveratrol. The kinetic experiments reveal that in an aqueous medium the absorption of resveratrol by the biomembranes models is allowed, whereas the absorption of its derivatives is hindered; in contrast when a lipophilic medium is employed, all three compounds are easily absorbed. PMID- 17397184 TI - Evaluation of metals in a defined medium for Pichia pastoris expressing recombinant beta-galactosidase. AB - Culture growth and recombinant protein yield of the Pichia pastoris GS115 methanol utilization positive system were studied in response to the types and levels of metals present in the growth medium and the supplemental salts typically used for these fermentations. Magnesium and zinc were both required to support cell growth but at significantly reduced levels compared to the control. However, supplementation with calcium, cobalt, iron, manganese, iodine, boron, and molybdenum were not required to sustain cell mass. When the medium was reformulated with only zinc and magnesium, the cells grew to 12-15 generations, which are expected for high cell density fed-batch fermentations. Product yields of the recombinant protein beta-galactosidase were significantly influenced by the trace metal concentrations. By using response surface and full factorial designs, maximum protein yield occurred when the concentration of zinc salt was limited to the level necessary only to support cell mass while protein yield positively correlated to increasing levels of the remaining trace metal salts. These studies are the first to show that excess trace metals must be optimized when developing P. pastoris based fed-batch fermentations. PMID- 17397185 TI - Heterologous GPCR expression: a bottleneck to obtaining crystal structures. AB - G protein-coupled receptors (GPCRs) are an important, medically relevant class of integral membrane proteins. Laboratories throughout all disciplines of science devote time and energy into developing practical methods for the discovery, isolation, and characterization of these proteins. Since the crystal structure of rhodopsin was solved 6 years ago, the race to determine high-resolution structures of more GPCRs has gained momentum. Since certain GPCRs are currently produced at sufficient levels for X-ray crystallography trials, it is speculated that heterologous expression of GPCRs may no longer be a bottleneck in obtaining crystal structures. This Review focuses on the current approaches in heterologous expression of GPCRs and explores the problems associated with obtaining crystal structures from GPCRs expressed in different systems. Although milligram amounts of certain GPCRs are attainable, the majority of GPCRs are still either produced at very low levels or not at all. Developing reliable expression techniques for GPCRs is still a major priority for the structural characterization of GPCRs. PMID- 17397186 TI - Upregulation of the immediate early gene product RhoB by exoenzyme C3 from Clostridium limosum and toxin B from Clostridium difficile. AB - ADP-ribosylation of Rho(A,B,C) by the family of exoenzyme C3-like transferases induces reorganization of the actin cytoskeleton based on inactivation of RhoA. No data are available on the role of RhoB in C3-treated cells. In murine fibroblasts treated with the cell-permeable exoenzyme C3 from Clostridium limosum (C3), an increase in the level of RhoB was observed. This upregulation of RhoB was based on transcriptional activation, as it was responsive to inhibition by actinomycin D and accompanied by activation of the rhoB promoter. Upregulation of RhoB was not observed in cells treated with either the actin ADP-ribosylating C2 toxin from Clostridium botulinum or latrunculin B, suggesting that inactivation of Rho but not actin reorganization was required for the upregulation of RhoB. This notion was confirmed, as the Rho/Rac/Cdc42-glucosylating toxin B from Clostridium difficile (TcdB) but not the Rac/R-Ras-glucosylating variant toxin B from C. difficile strain 1470 serotype F (TcdBF) induced a strong upregulation of RhoB. Upregulation of RhoB was further observed in response to the Rac/(H-,K-,N ,R-)Ras-glucosylating lethal toxin from Clostridium sordellii. The level of active, GTP-bound RhoB was increased in TcdB-treated cells compared to untreated cells (as determined by Rhotekin pull-down assay). In contrast, no active RhoB was found in C3-treated cells. RhoB-GTP was required for the TcdB-induced apoptosis (cytotoxic effect), as this effect was responsive to inhibition by C3. In conclusion, RhoB was upregulated by Rho-/Ras-inactivating toxins, as a consequence of the inactivation of either Rho(A,B,C) or (H-,K-,N-)Ras. In TcdB treated cells, RhoB escaped its inactivation and was required for the cytotoxic effect. PMID- 17397187 TI - Molecular dynamics simulation of the cytosolic mouth in Kcv-type potassium channels. AB - The functional effect of mutations near the intracellular mouth of the short viral Kcv potassium channel was studied by molecular dynamics simulations. As a model system we used the analogously mutated and truncated KirBac1.1, a channel with known crystal structure that shares genuine local sequence motifs with Kcv. By a novel simulated annealing methodology for structural averaging, information about the structure and dynamics of the intracellular mouth was extracted and complemented by Poisson-Boltzmann and 3D-RISM (reference interaction site model) integral equation theory for the determination of the K+ free energy surface. Besides the wild-type analogue of Kcv with its experimental reference activity (truncated KirBac1.1), two variants were studied: a deletion mutant where the N terminus is further truncated by eight amino acids, showing inactivity in the Kcv reference system, and a point mutant where the kink-forming proline at position 13 is substituted by alanine, resulting in hyperactivity. The computations reveal that the change of activity is closely related to a hydrophilic intracellular constriction formed by the C-terminal residues of the monomers. Hyperactivity of the point mutant is correlated with both sterical and electrostatic factors, while inactivity of the deletion mutant is related to a loss of specific salt bridge patterns between the C- and N-terminus at the constriction and to the consequences for ion passage barriers, as revealed by integral equation theory. The cytosolic gate, however, is probably formed by the N-terminal segment up to the proline kink and not by the constriction. The results are compared with design principles found for other channels. PMID- 17397188 TI - Mechanism of EF-Ts-catalyzed guanine nucleotide exchange in EF-Tu: contribution of interactions mediated by helix B of EF-Tu. AB - Elongation factor Tu (EF-Tu) belongs to the family of GTP-binding proteins and requires elongation factor Ts (EF-Ts) for nucleotide exchange. Crystal structures suggested that one of the salient features in the EF-Tu x EF-Ts complex is a conformation change in the switch II region of EF-Tu that is initiated by intrusion of Phe81 of EF-Ts between His84 and His118 of EF-Tu and may result in a destabilization of Mg2+ coordination and guanine nucleotide release. In the present paper, the contribution of His84 to nucleotide release was studied by pre steady-state kinetic analysis of nucleotide exchange in mutant EF-Tu in which His84 was replaced by Ala. Both intrinsic and EF-Ts-catalyzed nucleotide release was affected by the mutation, resulting in a 10-fold faster spontaneous GDP release and a 4-fold faster EF-Ts-catalyzed release of GTP and GDP. Removal of Mg2+ from the EF-Tu x EF-Ts complex increased the rate constant of GDP release 2 fold, suggesting a small contribution to nucleotide exchange. Together with published data on the effects of mutations interfering with other putative interactions between EF-Tu and EF-Ts, the results suggest that each of the contacts in the EF-Tu x EF-Ts complex alone contributes moderately to nucleotide destabilization, but together they act synergistically to bring about the overall 60,000-fold acceleration of nucleotide exchange in EF-Tu by EF-Ts. PMID- 17397189 TI - Trans-translation: the tmRNA-mediated surveillance mechanism for ribosome rescue, directed protein degradation, and nonstop mRNA decay. AB - The accurate flow of genetic information from DNA to RNA to protein is essential for all living organisms. An astonishing array of quality-assurance mechanisms have evolved to ensure that high degree of fidelity is maintained at every stage of this process. One of the most fascinating quality-control mechanisms involves tmRNA, also known as SsrA or 10Sa RNA. tmRNA is a versatile and highly conserved bacterial molecule endowed with the combined structural and functional properties of both a tRNA and a mRNA. The tmRNA system orchestrates three key biological functions: (1) recognition and rescue of ribosomes stalled on aberrant mRNAs, (2) disposal of the causative defective mRNAs, and (3) addition of a degradation tag to ribosome-associated protein fragments for directed proteolysis. Although not essential in Escherichia coli, tmRNA activity is essential for bacterial survival under adverse conditions and for virulence in some, and perhaps all, pathogenic bacteria. Recent evidence suggests that in addition to its quality-control function the tmRNA system might also play a key regulatory role in certain physiological pathways. This review will focus on recent advances in our understanding of the structural properties, mechanistic details, and physiological significance of this unique RNA and its principal protein partners. PMID- 17397190 TI - A convenient gHMQC-based NMR assay for investigating ammonia channeling in glutamine-dependent amidotransferases: studies of Escherichia coli asparagine synthetase B. AB - X-ray crystal structures of glutamine-dependent amidotransferases in their "active" conformation have revealed the existence of multiple active sites linked by solvent inaccessible intramolecular channels, giving rise to the widely accepted view that ammonia released in a glutaminase site is channeled efficiently into a separate synthetase site where it undergoes further reaction. We now report a very convenient isotope-edited 1H NMR-based assay that can be used to probe the transfer of ammonia between the active sites of amidotransferases and demonstrate its use in studies of Escherichia coli asparagine synthetase B (AS-B). Our NMR results suggest that (i) high glutamine concentrations do not suppress ammonia-dependent asparagine formation in this bacterial asparagine synthetase and (ii) ammonia in bulk solution can react with the thioester intermediate formed during the glutaminase half-reaction by accessing the N-terminal active site of AS-B during catalytic turnover. These observations are consistent with a model in which exogenous ammonia can access the intramolecular tunnel in AS-B during glutamine-dependent asparagine synthesis, in contrast to expectations based on studies of class I amidotransferases. PMID- 17397191 TI - Modeling of the complex between transducin and photoactivated rhodopsin, a prototypical G-protein-coupled receptor. AB - Obtaining a reliable 3D model for the complex formed by photoactivated rhodopsin (R*) and its G-protein, transducin (Gtalphabetagamma), would significantly benefit the entire field of structural biology of G-protein-coupled receptors (GPCRs). In this study, we have performed extensive configurational sampling for the isolated C-terminal fragment of the alpha-subunit of transducin, Gtalpha 340 350, within cavities of photoactivated rhodopsin formed by different energetically feasible conformations of the intracellular loops. Our results suggested a new 3D model of the rhodopsin-transducin complex that fully satisfied all available experimental data on site-directed mutagenesis of rhodopsin and Gtalphabetagamma as well as data from disulfide-linking experiments. Importantly, the experimental data were not used as a priori constraints in model building. We performed a thorough comparison of existing computational models of the rhodopsin transducin complex with each other and with current experimental data. It was found that different models suggest interactions with different molecules in the rhodopsin oligomer, that providing valuable guidance in design of specific novel experimental studies of the R*-Gtalphabetagamma complex. Finally, we demonstrated that the isolated Gtalpha 340-350 fragment does not necessarily bind rhodopsin in the same binding mode as the same segment in intact Gtalpha. PMID- 17397192 TI - Near-UV photolysis of 2-methyl-1,4-naphthoquinone-DNA duplexes: characterization of reversible and stable interstrand cross-links between quinone and adenine moieties. AB - Near-UV photolysis of 2-methyl-1,4-naphthoquinone (MQ, menadione) tethered DNA induces initial charge transfer followed by either transport of the damage to G or the formation of interstrand cross-links between MQ and DNA bases. In this work, the products responsible for interstrand cross-links have been characterized by mass spectrometry, NMR, and comparison with model compounds. Three major products were formed in the photolysis of MQ-DNA duplexes. Two of the products (isomers) have a cross-link between C2 of a 2,3-saturated MQ moiety and N6 of a dAdo moiety. These products readily convert back to MQ and dAdo upon heating in neutral solution, and thus, they lead to reversible cross-links in MQ DNA duplexes. The third product has a cross-link between C3 of a 2,3-unsaturated MQ moiety and N6 of an dAdo moiety. This product was stable in neutral solution. The formation of MQ to A cross-links in DNA may be explained by the coupling of MQ radicals that arise from the protonation of MQ radical anions, together with adenin-N6-yl radicals that arise from the deprotonation of A radical cations. PMID- 17397193 TI - Supramolecular sequential assembly of polymer thin films based on dimeric, dendrimeric, and polymeric Schiff-base ligands and metal ions. AB - New metal-Schiff-base coordination polymer films were prepared using multiple sequential adsorption of metal ions and salen-based ligand molecules. As the ligands, bis-bidentate 5,5'-methylene-bis(N-methylsalicylidenamine) (MBSA), tetra bidentate N,N',N' ',N' ''-tetrasalicylidene-polyamidoamine (TSPA), and multi bidentate poly(N-salicylidenevinylamine) (PSVA) were used. The metal ions were Cu(II), Zn(II), Fe(II), Fe(III), and Ce(IV). The resulting films are deeply colored due to the formation of coordinative bonds between the metal ions and the salen groups. Our study indicates that film formation becomes progressively easier, if the number of salen groups per ligand molecule increases. While Cu(II), Ni(II), Fe, and Ce(IV) are well suited for complex formation, Zn(II) is less suited. Possible structures of the polymers are discussed. Cyclic voltammetric studies of the films are also presented. PMID- 17397194 TI - AFM nanoindentations of diatom biosilica surfaces. AB - Diatoms have intricately and uniquely nanopatterned silica exoskeletons (frustules) and are a common target of biomimetic investigations. A better understanding of the diatom frustule structure and function at the nanoscale could provide new insights for the biomimetic fabrication of nanostructured ceramic materials and lightweight, yet strong, scaffold architectures. Here, we have mapped the nanoscale mechanical properties of Coscinodiscus sp. diatoms using atomic force microscopy (AFM)-based nanoindentation. Mechanical properties were correlated with the frustule structures obtained from high-resolution AFM and scanning electron microscopy (SEM). Significant differences in the micromechanical properties for the different frustule layers were observed. A comparative study of other related inorganic material including porous silicon films and free-standing membranes as well as porous alumina was also undertaken. PMID- 17397195 TI - Multiscale modeling of self-assembled monolayers of thiophenes on electronic material surfaces. AB - Computer simulation programs, spanning different time and length scales, are used to describe the fundamentals of thin film growth morphology in organic self assembled monolayers using thiophenes on gold as representative systems. Ab initio calculations created a catalog of the energetics between two N-[4-(thien 2ylethynyl)phenyl] hydroxyl ("1P" molecules) in vacuum and interactions in three orthogonal orientations (parallel, perpendicular, and gamma-phase) to a Au (111) surface. This energetic dataset was supplied as the input for kinetic Monte Carlo simulations of dimer and trimer representations of small organic molecules to describe both sub-monolayer and multilayer growth on a series of hypothetical model substrates. On strongly binding metallic-like substrates, sub-monolayers of the model organic molecules formed ordered phases in the x and y directions at high temperatures and a disordered polycrystalline structure at low temperatures with the molecules lying down. Only at high temperatures was a "phase inversion" observed from a completely flat to an upright structure, suggesting the upright phase to be kinetically limited. Results for multilayer deposition of 1P molecules on three substrates which differ in their binding energy to the molecule (from non-interacting to strongly binding substrates) provided a rich view of the polymorphism that can result from differing choices of temperature and flux conditions. Irrespective of the binding energy of the molecule to the substrate, on highly corrugated surfaces we always observed 3D-island growth of multiple layers of the thiophenes, in contrast to Stranski-Krastanov or Frank-van der Merwe growth on more uniform substrates. The qualitative picture we obtained agrees with the growth habits of other small organic molecule systems like the acene series. Finally, molecular dynamics studies were used to understand the packing structures of stable polymorphs of thiophene SAMs. Different deposition conditions and substrate-molecule binding captured different regimes of growth morphology, some of which have already been observed experimentally. PMID- 17397196 TI - Controlled stacking of charged block copolymer micelles. AB - Using poly(acrylic acid)-b-poly(methyl acrylate)-b-polystyrene (PAA-b-PMA-b-PS) triblock copolymers or a mixture of different molecular weight PAA-b-PS diblock copolymers, stacks of polymeric micellar assemblies, such as disks and Y-shaped cylinders, were formed through intermicellar interactions. Whereas micelles hierarchically stacked together, micellar interactions within the stack defined a uniform micelle geometry and size for up to micrometers in length. The kinetic pathway dependence and stability of the stacked assemblies were studied, and possible intermicellar interactions between micelles within the stacks are proposed. PMID- 17397197 TI - Micro- to nanoscale structure of biocompatible PLA-PEO-PLA hydrogels. AB - We observe large-scale structures in hydrogels of poly(l-lactide)-poly(ethylene oxide)-poly(l-lactide) (PLLA-PEO-PLLA) ranging in size from a few hundred nanometers to several micrometers. These structures are apparent through both ultra-small angle scattering (USAS) techniques and confocal microscopy. The hydrogels showed power law scattering in the USAS regime, which is indicative of scattering from fractal structures. The fractal dimension of the scattering from hydrogels revealed that the gels have large size aggregates with a mass fractal structure over the nanometer-to-micrometer length scales. The aggregates also seem to become more "dense" with an increase in the molecular weight of crystalline PLLA domains. Visualization through confocal microscopy confirms that the gels have a microstructure of interspersed micrometer-sized polymer inhomogeneities with water channels running between them. The presence of micrometer-sized water channels in the hydrogels has very important implications for biomedical applications. PMID- 17397198 TI - Surface organization, light-driven surface changes, and stability of semifluorinated azobenzene polymers. AB - A series of polymers with 4-perfluoroalkyl-modified azobenzene side groups was investigated for its light-induced changes in surface properties. The ultraviolet (UV) light activated trans to cis isomerization of the azobenzene group, and the influence of molecular order and orientation on this process were studied using near-edge X-ray absorption fine structure (NEXAFS) spectroscopy and water contact angle measurements. Light-induced molecular reorganization in the near-surface region was studied by NEXAFS using in situ UV irradiation of polymer thin films. Differential scanning calorimetry and wide-angle X-ray scattering studies showed that sufficiently long fluoroalkyl groups formed well-ordered smectic mesophases in the bulk, as well as on the surface, which was evidenced by NEXAFS. The disruption of mesogen packing by photoisomerization was found to be influenced by the fluoroalkyl segment length. Surfaces with perfluorohexyl and perfluorooctyl groups that showed high orientational order were also highly resistant to light induced changes. In such cases, the trans-cis isomerization resulted in greater lowering of the azobenzene phenyl ring order parameters than the perfluoroalkyl order parameters. UV exposure caused reorientation of the phenyl rings of the azobenzene group, but the terminal perfluoroalkyl segments remained more or less ordered. PMID- 17397199 TI - Polyelectrolyte blend multilayer films: surface morphology, wettability, and protein adsorption characteristics. AB - We report the influence of polyelectrolyte (PE) multilayer films prepared from poly(styrene sulfonate)-poly(acrylic acid) (PSS-PAA) blends, deposited in alternation with poly(allylamine hydrochloride) (PAH), on film wettability and the adsorption behavior of the protein immunoglobulin G (IgG). Variations in the chemical composition of the PAH/(PSS-PAA) multilayer films, controlled by the PSS/PAA blend ratio in the dipping solutions, were used to systematically control film thickness, surface morphology, surface wettability, and IgG adsorption. Spectroscopic ellipsometry measurements indicate that increasing the PSS content in the blend solutions results in a systematic decrease in film thickness. Increasing the PSS content in the blend solutions also leads to a reduction in film surface roughness (as measured by atomic force microscopy), with a corresponding increase in surface hydrophobicity. Advancing contact angles (theta) range from 7 degrees for PAH/PAA films through to 53 degrees for PAH/PSS films. X-ray photoelectron spectroscopy measurements indicate that the increase in film hydrophobicity is due to an increase in PSS concentration at the film surface. In addition, the influence of added electrolyte in the PE solutions was investigated. Adsorption from PE solutions containing added salt favors PSS adsorption and results in more hydrophobic films. The amount of IgG adsorbed on the multilayer films systematically increased on films assembled from blends with increasing PSS content, suggesting strong interactions between PSS in the multilayer films and IgG. Hence, multilayer films prepared from blended PE solutions can be used to tune film thickness and composition, as well as wetting and protein adsorption characteristics. PMID- 17397200 TI - The properties of a binary mixture of nonionic surfactants in water at the water/air interface. AB - The behavior of mixed nonionic/nonionic surfactant solutions, that is, p-(1,1,3,3 tetramethylbutyl)phenoxy poly(ethylene glycol)s Triton X-100 (TX100) and Triton X 165 (TX165) have been studied by surface tension and density measurements. The obtained results of the surface tension measurements were compared with those calculated from the relations derived by Joos, Miller, and co-workers. From the comparison, it appeared that by using these two approaches the adsorption behavior of TX100 and TX165 mixtures at different mole fractions can be predicted. The negative deviation from the linear relationship between the surface tension and composition of TX100 and TX165 mixtures in the concentration range corresponding to that of the saturated monolayer at the interface, the values of the parameters of molecular interaction, the activity coefficients, as well as the excess Gibbs energy of mixed monolayer formation calculated on the basis of Rosen and Motomura approaches proved that there is synergism in the reduction of the surface tension of aqueous solutions of TX100 and TX165 mixture when saturation of the monolayer is achieved. The negative parameters of intermolecular interaction in the mixed micelle and calculations based on MT theory of Blankschtein indicate that there is also synergism in the micelle formation for TX100 and TX165 mixture. It was also found that the values of the standard Gibbs energy of adsorption and micellization for the mixture of these two surfactants, which confirm the synergetic effect, can be predicted on the basis of the proposed equations, which include the values of the mole fraction of surfactant and excess Gibbs energy TX100 and TX165 in the monolayer and micelle. PMID- 17397201 TI - Surfactant templating effects on the encapsulation of iron oxide nanoparticles within silica microspheres. AB - Hollow silica microspheres encapsulating ferromagnetic iron oxide nanoparticles were synthesized by a surfactant-aided aerosol process and subsequent treatment. The cationic surfactant cetyltrimethyl ammonium bromide (CTAB) played an essential role in directing the structure of the composite. Translation from mesoporous silica particles to hollow particles was a consequence of increased loading of ferric species in the precursor solution and the competitive partitioning of CTAB between silicate and ferric colloids. The hypothesis was that CTAB preferentially adsorbed onto more positively charged ferric colloids under acidic conditions. At a critical Fe/Si ratio, most of the CTAB was adsorbed onto ferric colloids and coagulated the colloids to form larger clusters. During the aerosol process, a silica shell was first formed due to the preferred silicate condensation on the gas-liquid interface of the aerosol droplet. Subsequent drying concentrated the ferric clusters inside the silica shell and resulted in a silica shell/ferric core particle. Thermal treatment of the core shell particle led to encapsulation of a single iron oxide nanoparticle inside each silica hollow microsphere. PMID- 17397202 TI - Quenching of molecular fluorescence on the surface of monolayer-protected gold nanoparticles investigated using place exchange equilibria. AB - The insertion of fluorescently labeled thiols into the protecting self-assembled monolayer on the surface of gold nanoparticles through place exchange reactions and the effects of this insertion on the photophysical properties of the fluorophores are investigated. Analysis of solution-phase fluorescence data using a dynamic equilibrium model yields the equilibrium constant for the place exchange equilibrium, as well as the relative fluorescence brightness of the fluorophores on the particle surface. In all cases we find a significant quenching of the fluorescence, and potential reasons for this quenching are discussed. In the case of these relatively small particles (4.5 nm diameter), the quenching appears to be mainly related to enhanced nonradiative deactivation pathways. The place exchange equilibrium constant reveals a reduced affinity of the fluorescently labeled thiols for insertion into the nonfluorescent alkylthiol monolayer (K(eq) approximately 0.2) compared to unlabeled alkylthiols. PMID- 17397203 TI - Inhibiting surface crystallization of amorphous indomethacin by nanocoating. AB - An amorphous solid (glass) may crystallize faster at the surface than through the bulk, making surface crystallization a mechanism of failure for amorphous pharmaceuticals and other materials. An ultrathin coating of gold or polyelectrolytes inhibited the surface crystallization of amorphous indomethacin (IMC), an anti-inflammatory drug and model organic glass. The gold coating (10 nm) was deposited by sputtering, and the polyelectrolyte coating (3-20 nm) was deposited by an electrostatic layer-by-layer assembly of cationic poly(dimethyldiallyl ammonium chloride) (PDDA) and anionic sodium poly(styrenesulfonate) (PSS) in aqueous solution. The coating also inhibited the growth of existing crystals. The inhibition was strong even with one layer of PDDA. The polyelectrolyte coating still permitted fast dissolution of amorphous IMC and improved its wetting and flow. The finding supports the view that the surface crystallization of amorphous IMC is enabled by the mobility of a thin layer of surface molecules, and this mobility can be suppressed by a coating of only a few nanometers. This technique may be used to stabilize amorphous drugs prone to surface crystallization, with the aqueous coating process especially suitable for drugs of low aqueous solubility. PMID- 17397204 TI - Determination of membrane domain size by fluorescence resonance energy transfer: effects of domain polydispersity and packing. AB - Fluorescence resonance energy transfer (FRET) is sensitive to lateral heterogeneity in multicomponent membranes. Recently, we developed a model that enables the extraction of domain size from time-resolved FRET data, and here we examine the effects of domain polydispersity and spatial ordering on the accuracy of domain size determination. The model is applied to Monte Carlo calculations of membranes containing polydisperse domains, either randomly or hexagonally packed for three probe-partitioning schemes and three domain surface coverages. Statistical analysis of the model fitting results supports the conclusion that time-resolved FRET is a robust and precise technique for determining the size of small membrane domains (98:2). Simple treatment of the latter (RS,S) beta-halo-tert-butanesulfinamides with KOH afforded the corresponding (RS,S)-N (tert-butylsulfinyl)aziridines in quantitative yields. On the contrary, its epimer, (RS,R)-N-(tert-butylsulfinyl)aziridine was synthesized by switchover of the reducing agent from NaBH4 to LiBHEt3. (RS,R)-N-(tert-Butylsulfinyl)aziridines were synthesized in good yields (up to 85%) and diastereoselectivity (up to 92:8) by reduction of (RS)-N-tert-butanesulfinyl alpha-halo imines with LiBHEt3 in dry THF and subsequent treatment with KOH. All chiral aziridines were obtained as a single diastereomer after recrystallization (overall yield up to 91%) or after flash chromatography. PMID- 17397223 TI - Orientational isomerism controlled by the difference in electronic environments of a self-assembling heterodimeric capsule. AB - Tetrakis(4-hydroxyphenyl)-cavitand 1 and tetra(4-pyridyl)-cavitand 2 self assemble into a heterodimeric capsule 1.2 via four ArOH...pyridyl hydrogen bonds in CDCl3. The 1.2 expresses the orientational isomerism of an encapsulated unsymmetrical guest with high orientational selectivity because the electronic environment of the 1 unit is different from that of the 2 unit. For p ethoxyiodobenzene and 2-iodo-6-methoxynaphthalene encapsulated in 1.2, the iodo group is specifically oriented to the cavity of the 2 unit. The orientational isomeric selectivity for methyl p-acetoxybenzoate and methyl p-ethoxybenzoate within 1.2 is 1:0.11 and 1:<0.05, respectively, wherein the methyl ester group is preferentially oriented to the cavity of the 2 unit. The delicate balance among electrostatic potential repulsion, CH-pi interaction, or CH-halogen (halogen-pi) interaction, in 1.2-guest assembly influences the orientational isomeric selectivity of unsymmetrical guests within 1.2. PMID- 17397224 TI - Novel synthons from mucochloric acid: the first use of alpha,beta-dichloro-gamma butenolides and gamma-butyrolactams for direct vinylogous aldol addition. AB - Novel 5-(1'-hydroxy)-gamma-butyrolactone and gamma-butyrolactam subunits were synthesized by direct vinylogous aldol addition of alpha,beta-dichloro gamma butyrolactones and gamma-butyrolactams with aldehydes under basic conditions. Different bases and solvents were screened in the context of generating gamma butyrolactones. Diastereoselectivity was observed and gamma-butenolides and gamma butyrolactams showed opposite diastereoselectivity under the same reaction condition. PMID- 17397225 TI - Preparation of the 5/5-spiroketal of the ritterazines. AB - The enantiomerically pure 5/5-spiroketal required for the synthesis of the ritterazines has been prepared with high diastereocontrol by ring closure followed by equilibration. PMID- 17397226 TI - Practical syntheses of pyridinolines, important amino acidic biomarkers of collagen health. AB - The paper reports some successful results on the first fully stereoselective total synthesis of the collagen cross-link pyridinolines. All stereogenic centers are stereoselectively introduced using Williams glycine template methodology, and oxazinones are used as a source of chirality and as easily removable protecting groups of the amino acidic functionalities during the assembly of the pyridinoline nucleus. PMID- 17397227 TI - Parallel synthesis of a novel C2-aryl pyrrolo[2,1-c][1,4]benzodiazepine (PBD) library. AB - A 66-member C2-aryl pyrrolo[2,1-c][1,4]benzodiazepine (PBD) library has been successfully synthesized in parallel via Suzuki coupling using PS-PPh3Pd (catalyst) and PS-DEAM (scavenger) under microwave radiation. Library members were obtained in sufficient yield (up to 91%) and purity (85-98% crude) for biological evaluation. PMID- 17397228 TI - From the one-bead-one-compound concept to one-bead-one-reactor. AB - The one-bead-one-compound method gives access to millions of compounds that can be screened directly on the bead. Although characterization techniques are increasingly potent and reliable, problems can still be encountered in deciphering the sequence of the active compound because of sensitiveness or manipulation of the bead. ChemMatrix, a totally PEG-based resin, has resolved the synthesis of peptides of outstanding difficulty. Like other PEG-based resins, it permits on-bead screening because of its compatibility in aqueous media and has the further advantage of having a high loading, comparable to polystyrene resins. ChemMatrix beads previously swelled in water can be nicely divided into four parts that can be characterized using different analytical techniques or just stored for safety or for further testing. The four bead parts show high homogeneity and can thus be considered to be replicas. PMID- 17397230 TI - Tuning the mechanical properties of SWNT/nylon 6,10 composites with flexible spacers at the interface. AB - We have prepared nylon 6,10 nanocomposites using functionalized single wall carbon nanotubes and our interfacial in situ polycondensation method. The specific functional groups -(CH2)nCOCl [n = 4 and 9] on the sidewalls of SWNT were designed to covalently link nanotubes to the nylon matrix via alkyl segments. The composites with functionalized SWNT show significant improvements in tensile modulus, strength, and toughness relative to nylon and nylon modified with non-functionalized SWNT. The alkyl linkages at the SWNT/nylon 6,10 interface contribute significantly to improving the toughness of the composites. PMID- 17397229 TI - Efficient use of Ellman safety-catch linker for solid-phase assisted synthesis of multivalent glycoconjugates. PMID- 17397231 TI - Dipole-dipole interactions in nanoparticle superlattices. AB - Nanoparticles often self-assemble into hexagonal-close-packed (hcp) structures although it is predicted to be less stable than face-centered-cubic (fcc) packing in hard-sphere models. In addition to close-packed fcc and hcp superlattices, we observe formation of nonclose-packed simple-hexagonal (sh) superlattices of nearly spherical PbS, PbSe, and gamma-Fe2O3 nanocrystals. This surprisingly rich phase diagram of monodisperse semiconducting nanoparticles is explained by considering the interactions between nonlocal dipoles of individual nanoparticles. By calculating the total electrostatic and dispersive energies, we explain stability of the hcp and sh nanoparticle superlattices, introduce the superlattice phase diagram, and predict antiferroelectric ordering in dipolar nanoparticle superlattices. PMID- 17397232 TI - Local field asymmetry drives second-harmonic generation in non-centrosymmetric nanodimers. AB - We demonstrate that second-harmonic generation (SHG) from arrays of non centrosymmetric T-shaped gold nanodimers with a nanogap arises from asymmetry in the local fundamental field distribution and is not related strictly to nanogap size. Calculations show that the local field contains orthogonal polarization components not present in the exciting field, which yield the dominant SHG response. The strongest SHG responses occur through the local surface susceptibility of the particles for a fundamental field distributed asymmetrically at the particle perimeters. Weak responses result from more symmetric distributions despite high field enhancement in the nanogap. Nearly constant field enhancement persists for relatively large nanogap sizes. PMID- 17397233 TI - Breakdown of the continuum stokes-einstein relation for nanoparticle diffusion. AB - Cadmium selenide nanoparticles are found to diffuse approximately 200 times faster in a polymeric liquid than predicted by the Stokes-Einstein relation. This remarkable behavior is hypothesized to be due to the nanoparticles being smaller than the entanglement mesh to create a frictional drag that does not follow continuum expectations, in line with a theoretical calculation presented before. This is one of the first demonstrations of X-ray photo correlation spectroscopy applied to polymeric liquids, which we use to explain the simultaneous 60% viscosity reduction of the mixture through a proposed constraint release mechanism. PMID- 17397234 TI - Mechanisms of quantum dot energy engineering by metalorganic vapor phase epitaxy on patterned nonplanar substrates. AB - A novel technique for tuning the strength of quantum confinement in site controlled semiconductor quantum dots (QDs) is introduced and investigated theoretically and experimentally. The method makes use of controlled local growth rates during metalorganic vapor phase epitaxy on patterned arrays of inverted pyramids. A model accounting for precursor migration and adatom incorporation predicts the tuning in QD thickness as a function of the pattern parameters. The results are in good agreement with experimental findings. This technique offers means for designing QD photonic structures with potential applications in QD based cavity quantum electrodynamics and quantum information processing. PMID- 17397235 TI - Dynamics of size-selected gold nanoparticles studied by ultrafast electron nanocrystallography. AB - We report the studies of ultrafast electron nanocrystallography on size-selected Au nanoparticles (2-20 nm) supported on a molecular interface. Reversible surface melting, melting, and recrystallization were investigated with dynamical full profile radial distribution functions determined with subpicosecond and picometer accuracies. In an ultrafast photoinduced melting, the nanoparticles are driven to a nonequilibrium transformation, characterized by the initial lattice deformations, nonequilibrium electron-phonon coupling, and, upon melting, the collective bonding and debonding, transforming nanocrystals into shelled nanoliquids. The displasive structural excitation at premelting and the coherent transformation with crystal/liquid coexistence during photomelting differ from the reciprocal behavior of recrystallization, where a hot lattice forms from liquid and then thermally contracts. The degree of structural change and the thermodynamics of melting are found to depend on the size of nanoparticle. PMID- 17397236 TI - Enhancing the electrical and optoelectronic performance of nanobelt devices by molecular surface functionalization. AB - By functionalizing the surfaces of ZnO nanobelts (NBs) with a thin self-assembled molecular layer, the electrical and optoelectronic performances of a single NB based device are drastically improved. For a single NB-based device, due to energy band tuning and surface modification, the conductance was enhanced by 6 orders of magnitude upon functionalization; a coating molecule layer has changed a Schottky contact into an Ohmic contact without sophisticated deposition of multilayered metals. A functionalized NB showed negative differential resistance and exhibited huge improved photoconductivity and gas sensing response. The functionalized molecular layer also greatly reduced the etching rate of the ZnO NBs by buffer solution, largely extending their life time for biomedical applications. Our study demonstrates a new approach for improving the physical properties of oxide NBs and nanowires for device applications. PMID- 17397237 TI - Prediction and mechanistic interpretation of human oral drug absorption using MI QSAR analysis. AB - Membrane-interaction [MI]-QSAR analysis, which includes descriptors explicitly derived from simulations of solutes [drugs] interacting with phospholipid membrane models, was used to construct QSAR models for human oral intestinal drug absorption. A data set of 188 compounds, which are mainly drugs, was divided into a parent training set of 164 compounds and a test set of 24 compounds. Stable, but not highly fit [R2 = 0.68] MI-QSAR models could be built for all 188 compounds. However, the relatively large number [47] of drugs having 100% absorption, as well as all zwitterionic compounds [11], had to be eliminated from the training set in order to construct a linear five-term oral absorption diffusion model for 106 compounds which was both stable [R2 = 0.82, Q2 = 0.79] and predictive given the test set compounds were predicted with nearly the same average accuracy as the compounds of the training set. Intermolecular membrane solute descriptors are essential to building good oral absorption models, and these intermolecular descriptors are displaced in model optimizations and intramolecular solute descriptors found in published oral absorption QSAR models. A general form for all of the oral intestinal absorption MI-QSAR models has three classes of descriptors indicative of three thermodynamic processes: (1) solubility and partitioning, (2) membrane-solute interactions, and (3) flexibility of the solute and/or membrane. The intestinal oral absorption MI-QSAR models were compared to MI-QSAR models previously developed for Caco-2 cell permeation and for blood-brain barrier penetration. The MI-QSAR models for all three of these ADME endpoints share several common descriptors, and suggest a common mechanism of transport across all three barriers. A further analysis of these three types of MI-QSAR models has been done to identify descriptor-term differences across these three models, and the corresponding differences in thermodynamic transport behavior of the three barriers. PMID- 17397238 TI - Effects of pH and polyanions on the thermal stability of fibroblast growth factor 20. AB - Fibroblast growth factor 20 (FGF20) is a member of the FGF family with potential for use in several different therapeutic categories. In this work, we provide the first structural characterization of FGF20 using a wide variety of approaches. Like other members of the FGF family, FGF20 appears to possess a beta-trefoil structure. The effect of pH on the conformation and thermal stability of FGF20 is evaluated using far-UV circular dichroism (CD), intrinsic and ANS fluorescence, and high-resolution derivative UV absorption spectroscopy. Empirical phase diagrams are constructed to describe the solution behavior of FGF20 over a wide pH and temperature range. The protein appears to be unstable at pH <5, with aggregation and precipitation observed during dialysis. A major heat-induced conformational change also causes aggregation and precipitation of FGF20 at elevated temperatures. The highest thermal stability is observed near neutral pH (Tm ~55 degrees C at pH 7). The effect of several high- and low-molecular mass polyanions on the thermal stability of FGF20 is also examined using CD, intrinsic fluorescence, and DSC analysis. Among these ligands, heparin exhibits the greatest stabilizing effect on FGF20, increasing the Tm by more than 10 degrees C. PMID- 17397239 TI - Peloruside A synergizes with other microtubule stabilizing agents in cultured cancer cell lines. AB - The microtubule stabilizing agent peloruside A binds to a unique site on the tubulin alpha,beta-heterodimer compared to taxoid site drugs such as paclitaxel (Taxol), docetaxel (Taxotere), epothilone A, and discodermolide. Because the binding sites differ, peloruside A may be able to synergize with these taxoid site drugs when added in combination to cultured cells. Ovarian carcinoma cells (1A9) and myeloid leukemic cells (HL-60) were treated with different concentrations of peloruside A and taxoid site drugs, both compounds given singly and in combination in the nanomolar range, and the antiproliferative activity, G2/M blocking potency, and microtubule stabilizing activity of the treatments assessed. Cell proliferation was monitored using the MTT cell proliferation assay, cell cycle block was determined by flow cytometry, and stabilization of the tubulin polymer was assessed by Western blotting for beta-tubulin distributions in supernatant and pellet fractions of cell lysates. A combination index (CI) was calculated from the equation CI = D1/Dx1 + D2/Dx2 in which D1 and D2 are the concentrations of drug 1 and drug 2 that in combination give the same response as drug 1 alone (Dx1) or drug 2 alone (Dx2). A CI of less than 1 indicates synergy, equal to 1, additivity, and greater than 1, antagonism. Confidence intervals for each CI value were obtained using a bootstrapping procedure. In cell proliferation assays, statistically significant synergy was found between peloruside A and paclitaxel and epothilone A. Combinations of these two taxoid site drugs, however, also showed synergy in their effects on cell proliferation. These results confirm that peloruside A, when added in combination with other microtubule stabilizing agents, acts synergistically to enhance the antimitotic action of the drugs, but also highlight the complexity of drug interactions in intact cells. PMID- 17397240 TI - Physicochemical characterization of molecular assemblies of miltefosine and amphotericin B. AB - This study describes the interactions between two amphiphilic molecules with antileishmanial activity, amphotericin B (AmB) and miltefosine [hexadecylphosphocholine (HePC)], the latter being effective by the oral route. The effect of HePC on the aggregation state of AmB in aqueous solution and the interactions between the two agents were monitored using absorption spectroscopy and circular dichroism. Structural characterization of the mixed aggregates formed in water by dynamic light scattering (DLS) and cryofracture electron microscopy was performed. At concentrations above its critical micelle concentration, HePC was shown to interact with AmB, leading to an increase in the proportion of AmB in its monomeric form as a result of a micellar solubilization mechanism with a capacity of 26 +/- 3 mmol of AmB solubilized/mol of HePC, that is, nearly 40 molecules of HePC per molecule of AmB in the mixed micelles. These were revealed as individual and spherical aggregates close to 10 nm in diameter by both electron microscopy and DLS. Such a micellar formulation provides a new AmB-based system which might be useful in delivering AmB orally for visceral leishmaniasis bitherapy. PMID- 17397242 TI - Comparative genomic hybridization arrays in clinical pathology: progress and challenges. AB - Array-based comparative genomic hybridization (array CGH) genome scanning is a powerful method for the global detection of gains and losses of genetic material in both congenital and neoplastic disorders. When used as a clinical diagnostic test, array CGH combines the whole genome perspective of traditional G-banded cytogenetics with the targeted identification of cryptic chromosomal abnormalities characteristic of fluorescence in situ hybridization (FISH). However, the presence of structural variants in the human genome can complicate analysis of patient samples, and array CGH does not provide morphologic information about chromosome structure, balanced translocations, or the actual chromosomal location of segmental duplications. Identification of such anomalies has significant diagnostic and prognostic implications for the patient. We therefore propose that array CGH should be used as a guide to the presence of genomic structural rearrangements in germline and tumor genomes that can then be further characterized by FISH or G-banding, depending on the clinical scenario. In this article, we share some of our experiences with diagnostic array CGH and discuss recent progress and challenges involved with the integration of array CGH into clinical laboratory medicine. PMID- 17397243 TI - Impact of pharmacogenomics on clinical practice in oncology. AB - Multiple drug strategies for many cancer types are now readily available and there is a clear need for tools to inform decision making on therapy selection. Although there is still a long way to go before pharmacogenomics achieves the goal of individualized selection of cancer treatment, promising progress is being made. Genetic testing for thiopurine methyltransferase (TPMT) variant alleles in patients prior to mercaptopurine administration, and for UGT1A1*28 in patients prior to administration of irinotecan therapy, along with the instigation of genotype-guided clinical trials (e.g. TYMS) are important advances in cancer pharmacogenomics. Markers for the toxicity and efficacy of many oncology drugs remain unknown; however, the examples highlighted here suggest progress is being made towards the incorporation of pharmacogenomics into clinical practice in oncology. PMID- 17397244 TI - Cardiovascular proteomics: past, present, and future. AB - With cardiovascular (CV)-related disorders accounting for the highest mortality rates in the world, affecting the quantity and quality of life of patients and creating an economic burden of prolonged therapeutic intervention, there is great significance in understanding the cellular and molecular alterations that influence the progression of these pathologies. The cellular genotype is regulated by the DNA component, whilst the cellular phenotype is influenced by the protein complement. By improving the understanding of the molecular mechanisms that influence the protein profile, the pathologies that influence the intrinsic functions of the CV system may be detected earlier or managed more efficiently. This is achievable with technologies encompassed by 'proteomics.' Proteomic investigations of CV diseases, including dilated cardiomyopathy (DCM), atherosclerosis, and ischemia/reperfusion (I/R) injury, have identified candidate proteins altered with the pathologic states, complementing past biochemical and physiologic observations. Whilst proteomics is still a relatively new discipline to be applied to the basic scientific investigation of CV diseases, it is emerging as a technique to screen for potential biomarkers in both tissues/cells and biologic fluids (biofluids), as well as to identify the targets of existing therapeutics. By enabling the separation of complex mixtures over numerous dimensions, exploiting the intrinsic properties of proteins, including charge state, molecular mass, and hydrophobicity, in addition to cellular location, the discrete alterations within the cell may be resolved. Proteomics has shown alterations to myofilament proteins including troponin I and myosin light chain, correlating with the reduction in contractility in the myocardium from DCM and I/R. The diverse cell types that coalesce to induce atherosclerotic plaque formation have been investigated both collectively and individually to elucidate the influence of the modifications to single cell types on the developing plaque as a whole. Proteomics has also been used to observe changes to biofluids occurring with these pathologies, a new potential link between basic science and clinical applications. The development of CV proteomics has helped to identify a number of possible protein candidates, and offers the potential to treat and diagnose CV disease more effectively in the future. PMID- 17397245 TI - Treatment heterogeneity in asthma: genetics of response to leukotriene modifiers. AB - Despite advances in treatment, asthma continues to be a significant health and economic burden. Although asthma cannot be cured, several drugs, including beta2 agonists, corticosteroids, and leukotriene (LT) modifiers, are well tolerated and effective in minimizing symptoms, improving lung function, and preventing exacerbations. However, inter-patient variability in response to asthma drugs limits their effectiveness. It has been estimated that 60-80% of this inter patient variability may be attributable to genetic variation. LT modifiers, in particular, have been associated with heterogeneity in response. These drugs exert their action by inhibiting the activity of cysteinyl leukotrienes (CysLTs), which are potent bronchoconstrictors and pro-inflammatory agents. Two classes of LT modifiers are 5-lipoxygenase (ALOX5) inhibitors (zileuton) and leukotriene receptor antagonists (LTRAs) [montelukast, pranlukast, and zarfirlukast]. LT modifiers can be used as alternatives to low-dose inhaled corticosteroids (ICS) in mild persistent asthma, as add-on therapy to low- to medium-dose ICS in moderate persistent asthma, and as add-on to high-dose ICS and a long-acting ss2 agonist in severe persistent asthma. At least six genes encode key proteins in the LT pathway: arachidonate 5-lipoxygenase (ALOX5), ALOX5 activating protein (ALOX5AP [FLAP]), leukotriene A4 hydrolase (LTA4H), LTC4 synthase (LTC4S), the ATP-binding cassette family member ABCC1 (multidrug resistance protein 1 [MRP1]), and cysteinyl leukotriene receptor 1 (CYSLTR1). Studies have reported that genetic variation in ALOX5, LTA4H, LTC4S, and ABCC1 influences response to LT modifiers. Plasma concentrations of LTRAs vary considerably among patients. Physio-chemical characteristics make it likely that membrane efflux and uptake transporters mediate the absorption of LTRAs into the systemic circulation following oral administration. Genes that encode efflux and uptake transport proteins harbor many variants that could influence the pharmacokinetics, and particularly the bioavailability, of LTRAs, and could contribute to heterogeneity in response. In the future, large, well designed clinical trials studying the pharmacogenetics of LT modifiers in diverse populations are warranted to determine whether a genetic signature can be developed that will accurately predict which patients will respond. PMID- 17397246 TI - Rapid detection of the DPYD IVS14+1G>A mutation for screening patients to prevent fluorouracil-related toxicity. AB - BACKGROUND: Deficiency of dihydropyrimidine dehydrogenase (DPD) has been linked to severe or lethal fluorouracil (FU)-related toxicity. The most prominent mutation in the DPYD gene is the IVS14+1G>A mutation, which causes skipping of exon 14 in the messenger RNA (mRNA) and results in DPD enzyme deficiency. Several methods have been described to detect this mutation, but all are labor intensive and low throughput. OBJECTIVE Our aim was to develop a high-throughput real-time PCR assay to screen patients for the IVS14+1G>A mutation. METHODS: Primers and probes were developed and several reaction conditions were tested. In total, 165 individuals were screened for this mutation, with DNA sequencing as a reference method. RESULTS: Results of the real-time PCR assay and DNA sequencing were 100% identical. In total, eight heterozygous individuals were identified, of which six were patients with severe FU-related toxicity after FU or capecitabine treatment and two were healthy volunteers. CONCLUSION: This new real-time PCR assay with a high throughput is particularly suitable for large-scale screening for the IVS14+1G>A mutation in patients selected for treatment with fluoropyrimidines in order to prevent severe FU-related toxicity. PMID- 17397247 TI - Rapid diagnosis of myocardial injury with troponin T and CK-MB relative index. AB - BACKGROUND: Current hospital practice involves protracted observation of chest pain patients to rule out myocardial infarction. Concurrent measurement of multiple biomarkers may increase sensitivity and make rapid diagnosis feasible. OBJECTIVE: We sought to determine the optimal biomarker strategy for highly sensitive, early diagnosis of myocardial injury. STUDY DESIGN: A prospective evaluation of 171 acute coronary syndrome patients admitted to a single university medical center was performed. Blood tests for creatine kinase (CK), CK myocardial band isoenzyme (CK-MB), and troponin T were obtained at 0, 3, 6, 8, and 16 hours after presentation to the emergency department. Myocardial injury was defined as a troponin T level of >or=0.03 ng/mL. RESULTS: Troponin T had sensitivities of 79.7%, 95.7%, and 98.4% at the time of initial presentation, 3 and 6 hours after presentation, respectively. Using a combination of troponin T and CK-MB relative index, sensitivity on presentation was increased to 90.6%. The sensitivity was improved to 97.9% and 100% at 3 and 6 hours, respectively. CONCLUSION: This study demonstrates that the diagnosis of myocardial injury can be accurately excluded within 6 hours of admission with high sensitivity using troponin T. The combination of troponin T and CK-MB relative index provided the largest improvement in diagnostic sensitivity at patient arrival. These results support the feasibility of rapid, efficient triage for the emergent presentation of patients with chest pain. PMID- 17397248 TI - Prenatal diagnosis of common aneuploidies in transcervical samples using quantitative fluorescent-PCR analysis. AB - AIM: The aim of this study was to test the feasibility of diagnosing common fetal chromosomal aneuploidies using quantitative fluorescent (QF)-PCR on transcervical cell (TCC) samples collected in the first trimester of pregnancy by means of intrauterine lavage (IUL). METHODS: A total of 181 TCC samples were retrieved from pregnant women between 5 and 12 weeks of gestation, immediately before elective termination of pregnancy, at which time corresponding placental tissue and maternal blood specimens were also obtained. Isolation of trophoblastic cells by micromanipulation was attempted in all TCC samples. Micromanipulated specimens were analyzed by multiplex QF-PCR, including short tandem repeats for the chromosomes X, Y, 21, 18, and 13. RESULTS: The micromanipulation was successful in 152 of 181 cases (84.8%) where chorionic villous filaments and/or cell clumps of seeming trophoblastic origin could be isolated. All 152 samples were tested by QF-PCR analysis and peaks of paternal origin could be documented in all cases. Two cases of trisomy 21 and two cases of monosomy X0 were detected by means of QF PCR assay, in accordance with the results obtained in corresponding placental samples. CONCLUSION: This study provides evidence that the use of multiplex QF PCR amplification of selected microsatellites could be applied to micromanipulated TCC samples and in particular to IUL samples, which often contain trophoblastic cells, for the detection of chromosomal aneuploidies. The approach described in this study appears, therefore, a very promising tool toward non-invasive prenatal genetic diagnosis in the early stage of gestation. PMID- 17397249 TI - The influence of NSAIDs on coumarin sensitivity in patients with CYP2C9 polymorphism after total hip replacement surgery. AB - OBJECTIVE: To determine the influence of NSAIDs on the international normalized ratio (INR) in patients with cytochrome P450 (CYP)-2C9 enzyme variants starting acenocoumarol (an oral coumarin) therapy during the first 7 days after total hip replacement surgery. METHODS: In this prospective study, an age-dependent protocol was used for the initiation of the acenocoumarol dose. Low-molecular weight heparin was given for 5 days. The study included 100 patients undergoing total hip replacement surgery. After the inclusion of the last patient, polymerase chain reaction CYP2C9 mutation testing was performed for all patients. Drug-use evaluation of NSAIDs and other potential coumarin-drug interactions was also performed. RESULTS: Eleven patients had an INR on 1 or more days >4.9. There were 52 patients who were using NSAIDs. Patients with a CYP2C9 mutation had a mean INR curve similar to patients without the mutation when NSAIDs were not coadministered. Within the group of patients heterozygous for a CYP2C9 mutation (n=30) only concomitant use of a NSAID resulted in an INR >4.9 (0% vs 38.9%, p<0.05). CONCLUSION: In the group of patients with a CYP2C9 variant (*2 or *3 alleles), only concomitant use of a NSAID resulted in INRs >4.9. The cost effectiveness of CYP2C9 screening before elective surgery has yet to be determined. PMID- 17397250 TI - MEN1 935-1G>C splicing mutation in an Indian patient with multiple endocrine neoplasia type 1. AB - BACKGROUND AND OBJECTIVE: Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant syndrome characterized mainly by multiple tumors involving parathyroid, pancreatic, and pituitary glands. To date, there have been no genetic studies reported on MEN1 in the Indian population. In order to begin to establish molecular diagnosis to improve the management of MEN1 in India, we performed a molecular analysis of the MEN1 gene in a patient of Indian origin. METHODS: Molecular analysis of the MEN1 gene was performed to identify mutations in an Indian patient previously diagnosed with sporadic MEN1. All the 10 exons of the MEN1 gene were amplified using the polymerase chain reaction and screened by direct DNA sequencing. RESULTS: The DNA sequencing results revealed the presence of an intronic, heterozygous, splicing mutation 935-1G>C in intron 5 of the MEN1 gene. CONCLUSION: This study provides the first data on genetic analysis of MEN1 in Indian patients. PMID- 17397251 TI - Deciphering protein-protein interactions. Part I. Experimental techniques and databases. PMID- 17397252 TI - Ten simple rules for a successful collaboration. PMID- 17397253 TI - Query-dependent banding (QDB) for faster RNA similarity searches. AB - When searching sequence databases for RNAs, it is desirable to score both primary sequence and RNA secondary structure similarity. Covariance models (CMs) are probabilistic models well-suited for RNA similarity search applications. However, the computational complexity of CM dynamic programming alignment algorithms has limited their practical application. Here we describe an acceleration method called query-dependent banding (QDB), which uses the probabilistic query CM to precalculate regions of the dynamic programming lattice that have negligible probability, independently of the target database. We have implemented QDB in the freely available Infernal software package. QDB reduces the average case time complexity of CM alignment from LN(2.4) to LN(1.3) for a query RNA of N residues and a target database of L residues, resulting in a 4-fold speedup for typical RNA queries. Combined with other improvements to Infernal, including informative mixture Dirichlet priors on model parameters, benchmarks also show increased sensitivity and specificity resulting from improved parameterization. PMID- 17397254 TI - Structural descriptors of gp120 V3 loop for the prediction of HIV-1 coreceptor usage. AB - HIV-1 cell entry commonly uses, in addition to CD4, one of the chemokine receptors CCR5 or CXCR4 as coreceptor. Knowledge of coreceptor usage is critical for monitoring disease progression as well as for supporting therapy with the novel drug class of coreceptor antagonists. Predictive methods for inferring coreceptor usage based on the third hypervariable (V3) loop region of the viral gene coding for the envelope protein gp120 can provide us with these monitoring facilities while avoiding expensive phenotypic tests. All simple heuristics (such as the 11/25 rule) as well as statistical learning methods proposed to date predict coreceptor usage based on sequence features of the V3 loop exclusively. Here, we show, based on a recently resolved structure of gp120 with an untruncated V3 loop, that using structural information on the V3 loop in combination with sequence features of V3 variants improves prediction of coreceptor usage. In particular, we propose a distance-based descriptor of the spatial arrangement of physicochemical properties that increases discriminative performance. For a fixed specificity of 0.95, a sensitivity of 0.77 was achieved, improving further to 0.80 when combined with a sequence-based representation using amino acid indicators. This compares favorably with the sensitivities of 0.62 for the traditional 11/25 rule and 0.73 for a prediction based on sequence information as input to a support vector machine and constitutes a statistically significant improvement. A detailed analysis and interpretation of structural features important for classification shows the relevance of several specific hydrogen-bond donor sites and aliphatic side chains to coreceptor specificity towards CCR5 or CXCR4. Furthermore, an analysis of side chain orientation of the specificity-determining residues suggests a major role of one side of the V3 loop in the selection of the coreceptor. The proposed method constitutes the first approach to an improved prediction of coreceptor usage based on an original integration of structural bioinformatics methods with statistical learning. PMID- 17397255 TI - Potential energy landscape and robustness of a gene regulatory network: toggle switch. AB - Finding a multidimensional potential landscape is the key for addressing important global issues, such as the robustness of cellular networks. We have uncovered the underlying potential energy landscape of a simple gene regulatory network: a toggle switch. This was realized by explicitly constructing the steady state probability of the gene switch in the protein concentration space in the presence of the intrinsic statistical fluctuations due to the small number of proteins in the cell. We explored the global phase space for the system. We found that the protein synthesis rate and the unbinding rate of proteins to the gene were small relative to the protein degradation rate; the gene switch is monostable with only one stable basin of attraction. When both the protein synthesis rate and the unbinding rate of proteins to the gene are large compared with the protein degradation rate, two global basins of attraction emerge for a toggle switch. These basins correspond to the biologically stable functional states. The potential energy barrier between the two basins determines the time scale of conversion from one to the other. We found as the protein synthesis rate and protein unbinding rate to the gene relative to the protein degradation rate became larger, the potential energy barrier became larger. This also corresponded to systems with less noise or the fluctuations on the protein numbers. It leads to the robustness of the biological basins of the gene switches. The technique used here is general and can be applied to explore the potential energy landscape of the gene networks. PMID- 17397256 TI - DNA familial binding profiles made easy: comparison of various motif alignment and clustering strategies. AB - Transcription factor (TF) proteins recognize a small number of DNA sequences with high specificity and control the expression of neighbouring genes. The evolution of TF binding preference has been the subject of a number of recent studies, in which generalized binding profiles have been introduced and used to improve the prediction of new target sites. Generalized profiles are generated by aligning and merging the individual profiles of related TFs. However, the distance metrics and alignment algorithms used to compare the binding profiles have not yet been fully explored or optimized. As a result, binding profiles depend on TF structural information and sometimes may ignore important distinctions between subfamilies. Prediction of the identity or the structural class of a protein that binds to a given DNA pattern will enhance the analysis of microarray and ChIP chip data where frequently multiple putative targets of usually unknown TFs are predicted. Various comparison metrics and alignment algorithms are evaluated (a total of 105 combinations). We find that local alignments are generally better than global alignments at detecting eukaryotic DNA motif similarities, especially when combined with the sum of squared distances or Pearson's correlation coefficient comparison metrics. In addition, multiple-alignment strategies for binding profiles and tree-building methods are tested for their efficiency in constructing generalized binding models. A new method for automatic determination of the optimal number of clusters is developed and applied in the construction of a new set of familial binding profiles which improves upon TF classification accuracy. A software tool, STAMP, is developed to host all tested methods and make them publicly available. This work provides a high quality reference set of familial binding profiles and the first comprehensive platform for analysis of DNA profiles. Detecting similarities between DNA motifs is a key step in the comparative study of transcriptional regulation, and the work presented here will form the basis for tool and method development for future transcriptional modeling studies. PMID- 17397257 TI - Hsp90 selectively modulates phenotype in vertebrate development. AB - Compromised heat shock protein 90 (Hsp90) function reveals cryptic phenotypes in flies and plants. These observations were interpreted to suggest that this molecular stress-response chaperone has a capacity to buffer underlying genetic variation. Conversely, the protective role of Hsp90 could account for the variable penetrance or severity of some heritable developmental malformations in vertebrates. Using zebrafish as a model, we defined Hsp90 inhibitor levels that did not induce a heat shock response or perturb phenotype in wild-type strains. Under these conditions the severity of the recessive eye phenotype in sunrise, caused by a pax6b mutation, was increased, while in dreumes, caused by a sufu mutation, it was decreased. In another strain, a previously unobserved spectrum of severe structural eye malformations, reminiscent of anophthalmia, microphthalmia, and nanophthalmia complex in humans, was uncovered by this limited inhibition of Hsp90 function. Inbreeding of offspring from selected unaffected carrier parents led to significantly elevated malformation frequencies and revealed the oligogenic nature of this phenotype. Unlike in Drosophila, Hsp90 inhibition can decrease developmental stability in zebrafish, as indicated by increased asymmetric presentation of anophthalmia, microphthalmia, and nanophthalmia and sunrise phenotypes. Analysis of the sunrise pax6b mutation suggests a molecular mechanism for the buffering of mutations by Hsp90. The zebrafish studies imply that mild perturbation of Hsp90 function at critical developmental stages may underpin the variable penetrance and expressivity of many developmental anomalies where the interaction between genotype and environment plays a major role. PMID- 17397258 TI - Off the beaten path: an interview with Spencer Wells by Jane Gitschier. PMID- 17397259 TI - The molecular basis of high-altitude adaptation in deer mice. AB - Elucidating genetic mechanisms of adaptation is a goal of central importance in evolutionary biology, yet few empirical studies have succeeded in documenting causal links between molecular variation and organismal fitness in natural populations. Here we report a population genetic analysis of a two-locus alpha globin polymorphism that underlies physiological adaptation to high-altitude hypoxia in natural populations of deer mice, Peromyscus maniculatus. This system provides a rare opportunity to examine the molecular underpinnings of fitness related variation in protein function that can be related to a well-defined selection pressure. We surveyed DNA sequence variation in the duplicated alpha globin genes of P. maniculatus from high- and low-altitude localities (i) to identify the specific mutations that may be responsible for the divergent fine tuning of hemoglobin function and (ii) to test whether the genes exhibit the expected signature of diversifying selection between populations that inhabit different elevational zones. Results demonstrate that functionally distinct protein alleles are maintained as a long-term balanced polymorphism and that adaptive modifications of hemoglobin function are produced by the independent or joint effects of five amino acid mutations that modulate oxygen-binding affinity. PMID- 17397260 TI - Systematic identification of cellular signals reactivating Kaposi sarcoma associated herpesvirus. AB - The herpesvirus life cycle has two distinct phases: latency and lytic replication. The balance between these two phases is critical for viral pathogenesis. It is believed that cellular signals regulate the switch from latency to lytic replication. To systematically evaluate the cellular signals regulating this reactivation process in Kaposi sarcoma-associated herpesvirus, the effects of 26,000 full-length cDNA expression constructs on viral reactivation were individually assessed in primary effusion lymphoma-derived cells that harbor the latent virus. A group of diverse cellular signaling proteins were identified and validated in their effect of inducing viral lytic gene expression from the latent viral genome. The results suggest that multiple cellular signaling pathways can reactivate the virus in a genetically homogeneous cell population. Further analysis revealed that the Raf/MEK/ERK/Ets-1 pathway mediates Ras-induced reactivation. The same pathway also mediates spontaneous reactivation, which sets the first example to our knowledge of a specific cellular pathway being studied in the spontaneous reactivation process. Our study provides a functional genomic approach to systematically identify the cellular signals regulating the herpesvirus life cycle, thus facilitating better understanding of a fundamental issue in virology and identifying novel therapeutic targets. PMID- 17397262 TI - Virus evolution reveals an exclusive role for LEDGF/p75 in chromosomal tethering of HIV. AB - Retroviruses by definition insert their viral genome into the host cell chromosome. Although the key player of retroviral integration is viral integrase, a role for cellular cofactors has been proposed. Lentiviral integrases use the cellular protein LEDGF/p75 to tether the preintegration complex to the chromosome, although the existence of alternative host proteins substituting for the function of LEDGF/p75 in integration has been proposed. Truncation mutants of LEDGF/p75 lacking the chromosome attachment site strongly inhibit HIV replication by competition for the interaction with integrase. In an attempt to select HIV strains that can overcome the inhibition, we now have used T-cell lines that stably express a C-terminal fragment of LEDGF/p75. Despite resistance development, the affinity of integrase for LEDGF/p75 is reduced and replication kinetics in human primary T cells is impaired. Detection of the integrase mutations A128T and E170G at key positions in the LEDGF/p75-integrase interface provides in vivo evidence for previously reported crystallographic data. Moreover, the complementary inhibition by LEDGF/p75 knockdown and mutagenesis at the integrase-LEDGF/p75 interface points to the incapability of HIV to circumvent LEDGF/p75 function during proviral integration. Altogether, the data provide a striking example of the power of viral molecular evolution. The results underline the importance of the LEDGF/p75 HIV-1 interplay as target for innovative antiviral therapy. Moreover, the role of LEDGF/p75 in targeting integration will stimulate research on strategies to direct gene therapy vectors into safe landing sites. PMID- 17397261 TI - Adaptation to human populations is revealed by within-host polymorphisms in HIV-1 and hepatitis C virus. AB - CD8(+) cytotoxic T-lymphocytes (CTLs) perform a critical role in the immune control of viral infections, including those caused by human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV). As a result, genetic variation at CTL epitopes is strongly influenced by host-specific selection for either escape from the immune response, or reversion due to the replicative costs of escape mutations in the absence of CTL recognition. Under strong CTL-mediated selection, codon positions within epitopes may immediately "toggle" in response to each host, such that genetic variation in the circulating virus population is shaped by rapid adaptation to immune variation in the host population. However, this hypothesis neglects the substantial genetic variation that accumulates in virus populations within hosts. Here, we evaluate this quantity for a large number of HIV-1- (n > or = 3,000) and HCV-infected patients (n > or = 2,600) by screening bulk RT-PCR sequences for sequencing "mixtures" (i.e., ambiguous nucleotides), which act as site-specific markers of genetic variation within each host. We find that nonsynonymous mixtures are abundant and significantly associated with codon positions under host-specific CTL selection, which should deplete within-host variation by driving the fixation of the favored variant. Using a simple model, we demonstrate that this apparently contradictory outcome can be explained by the transmission of unfavorable variants to new hosts before they are removed by selection, which occurs more frequently when selection and transmission occur on similar time scales. Consequently, the circulating virus population is shaped by the transmission rate and the disparity in selection intensities for escape or reversion as much as it is shaped by the immune diversity of the host population, with potentially serious implications for vaccine design. PMID- 17397263 TI - Type III effector activation via nucleotide binding, phosphorylation, and host target interaction. AB - The Pseudomonas syringae type III effector protein avirulence protein B (AvrB) is delivered into plant cells, where it targets the Arabidopsis RIN4 protein (resistance to Pseudomonas maculicula protein 1 [RPM1]-interacting protein). RIN4 is a regulator of basal host defense responses. Targeting of RIN4 by AvrB is recognized by the host RPM1 nucleotide-binding leucine-rich repeat disease resistance protein, leading to accelerated defense responses, cessation of pathogen growth, and hypersensitive host cell death at the infection site. We determined the structure of AvrB complexed with an AvrB-binding fragment of RIN4 at 2.3 A resolution. We also determined the structure of AvrB in complex with adenosine diphosphate bound in a binding pocket adjacent to the RIN4 binding domain. AvrB residues important for RIN4 interaction are required for full RPM1 activation. AvrB residues that contact adenosine diphosphate are also required for initiation of RPM1 function. Nucleotide-binding residues of AvrB are also required for its phosphorylation by an unknown Arabidopsis protein(s). We conclude that AvrB is activated inside the host cell by nucleotide binding and subsequent phosphorylation and, independently, interacts with RIN4. Our data suggest that activated AvrB, bound to RIN4, is indirectly recognized by RPM1 to initiate plant immune system function. PMID- 17397265 TI - Natural approaches to epilepsy. AB - This article reviews research on the use of diet, nutritional supplements, and hormones in the treatment of epilepsy. Potentially beneficial dietary interventions include identifying and treating blood glucose dysregulation, identifying and avoiding allergenic foods, and avoiding suspected triggering agents such as alcohol, aspartame, and monosodium glutamate. The ketogenic diet may be considered for severe, treatment-resistant cases. The Atkins diet (very low in carbohydrates) is a less restrictive type of ketogenic diet that may be effective in some cases. Nutrients that may reduce seizure frequency include vitamin B6, magnesium, vitamin E, manganese, taurine, dimethylglycine, and omega 3 fatty acids. Administration of thiamine may improve cognitive function in patients with epilepsy. Supplementation with folic acid, vitamin B6, biotin, vitamin D, and L-carnitine may be needed to prevent or treat deficiencies resulting from the use of anticonvulsant drugs. Vitamin K1 has been recommended near the end of pregnancy for women taking anticonvulsants. Melatonin may reduce seizure frequency in some cases, and progesterone may be useful for women with cyclic exacerbations of seizures. In most cases, nutritional therapy is not a substitute for anticonvulsant medications. However, in selected cases, depending on the effectiveness of the interventions, dosage reductions or discontinuation of medications may be possible. PMID- 17397264 TI - Bacterial ligands generated in a phagosome are targets of the cytosolic innate immune system. AB - Macrophages are permissive hosts to intracellular pathogens, but upon activation become microbiocidal effectors of innate and cell-mediated immunity. How the fate of internalized microorganisms is monitored by macrophages, and how that information is integrated to stimulate specific immune responses is not understood. Activation of macrophages with interferon (IFN)-gamma leads to rapid killing and degradation of Listeria monocytogenes in a phagosome, thus preventing escape of bacteria to the cytosol. Here, we show that activated macrophages induce a specific gene expression program to L. monocytogenes degraded in the phago-lysosome. In addition to activation of Toll-like receptor (TLR) signaling pathways, degraded bacteria also activated a TLR-independent transcriptional response that was similar to the response induced by cytosolic L. monocytogenes. More specifically, degraded bacteria induced a TLR-independent IFN-beta response that was previously shown to be specific to cytosolic bacteria and not to intact bacteria localized to the phagosome. This response required the generation of bacterial ligands in the phago-lysosome and was largely dependent on nucleotide binding oligomerization domain 2 (NOD2), a cytosolic receptor known to respond to bacterial peptidoglycan fragments. The NOD2-dependent response to degraded bacteria required the phagosomal membrane potential and the activity of lysosomal proteases. The NOD2-dependent IFN-beta production resulted from synergism with other cytosolic microbial sensors. This study supports the hypothesis that in activated macrophages, cytosolic innate immune receptors are activated by bacterial ligands generated in the phagosome and transported to the cytosol. PMID- 17397266 TI - Colds and influenza: a review of diagnosis and conventional, botanical, and nutritional considerations. AB - The common cold is the leading cause of doctor visits in the United States and annually results in 189 million lost school days. In the course of one year the U.S. population contracts approximately 1 billion colds. Influenza infection is still a leading cause of morbidity and mortality, accounting for 20-25 million doctor visits and 36,000 deaths per year in the United States. Conventional therapies for colds and flu focus primarily on temporary symptom relief and include over-the-counter antipyretics, anti-inflammatories, and decongestants. Treatment for influenza also includes prescription antiviral agents and vaccines for prevention. This article reviews the common cold and influenza viruses, presents the conventional treatment options, and highlights select botanicals (Echinacea spp., Sambucus nigra, larch arabinogalactan, Astragalus membranaceous, Baptisia tinctoria, Allium sativa, Panax quinquefolium, Eleutherococcus senticosus, Andrographis paniculata, olive leaf extract, and Isatis tinctoria) and nutritional considerations (vitamins A and C, zinc, high lactoferrin whey protein, N-acetylcysteine, and DHEA) that may help in the prevention and treatment of these conditions. PMID- 17397267 TI - Body temperature variability (Part 2): masking influences of body temperature variability and a review of body temperature variability in disease. AB - This is the second of a two-part review on body temperature variability. Part 1 discussed historical and modern findings on average body temperatures. It also discussed endogenous sources of temperature variability, including variations caused by site of measurement; circadian, menstrual, and annual biological rhythms; fitness; and aging. Part 2 reviews the effects of exogenous masking agents - external factors in the environment, diet, or lifestyle that can be a significant source of body temperature variability. Body temperature variability findings in disease states are also reviewed. PMID- 17397268 TI - Effect of various natural products on growth of bladder cancer cells: two promising mushroom extracts. AB - Despite the availability of several therapeutic options, a safer and more effective modality is urgently needed for treatment of bladder cancer. Specific immunotherapy is effective, but severe side effects limit its clinical use and underscore the need for unconventional therapies using less toxic substances. Many natural substances are touted for their medicinal aspects and side effect profiles, and some of these have been well characterized for their biological and medicinal properties. Accordingly, the effects on bladder cancer cells in vitro were investigated. Eight commercially available natural products were tested for possible effects on the growth of human bladder cancer T24 cells. This study demonstrated that two mushroom extracts, GD- and PL-fractions, induced a significant (>90 percent) growth reduction in 72 hours, whereas the remaining six products had no effect. Interestingly, non-toxic concentrations of the GD- or PL fractions, when combined with a non-toxic concentration of vitamin C, became highly cytotoxic, resulting in >90-percent cell death. Thus, vitamin C appears to act synergistically with these fractions to potentiate their bioactivity (cytotoxicity). No other products tested demonstrated such a synergistic potentiation with vitamin C. The present study indicates that GD- and PL fractions appear to have the most potent cytotoxic effect on human bladder cancer T24 cells. It is thus plausible that these substances could be used, solely or combined with conventional modalities, for the treatment of superficial bladder cancer. PMID- 17397269 TI - Centella asiatica. PMID- 17397270 TI - Biotin. PMID- 17397275 TI - Differences in risk factors for suicidality between African American and White patients vulnerable to suicide. AB - Risk factors for suicidal ideation and attempts have been shown to differ between African Americans and Whites across the lifespan. In the present study, risk factors for suicidality were examined separately by race/ethnicity in a population of 131 older adult patients considered vulnerable to suicide due to substance abuse and/or medical frailty. In adjusted analyses, social support was significantly associated with suicidality in African American patients, while younger age and the presence of an anxiety disorder were significantly associated with suicidality in White patients. The results suggest that race/ethnicity specific risk profiles may improve the detection of suicidality in vulnerable populations. PMID- 17397276 TI - Are Mexican American adolescents at greater risk of suicidal behaviors? AB - A reexamination of ethnicity as a risk factor for adolescent suicidal behavior, focusing on whether Mexican American youths are at increased risk, was undertaken. Data from a sample of 4,175 African, European, and Mexican Americans, aged 11-17, are presented. We examined lifetime attempts and past year attempts, thoughts, and plans. Odds ratios, adjusting for covariates, indicate no differences between European and Mexican Americans on past year thoughts, plans, or attempts or lifetime attempts. Although some studies have reported Mexican American youths are at increased risk, we did not find any differences. Possible explanations for disparate results across studies are discussed, in particular methods effects. PMID- 17397277 TI - Racial disparities in mental health service use by adolescents who thought about or attempted suicide. AB - Differences in rates and predictors of mental health service use among 2,226 Black, Hispanic, and White adolescents (aged 12-17) who reported recent suicidal thoughts or an attempt were examined. Black adolescents were 65% (OR = .65, p < .05), and Hispanic adolescents were 55% (OR = .55, p < .001), as likely as White adolescents to report service use, even when controlling for need for care and ability to secure services. Suicide attempt and psychiatric symptoms each interacted with race to increase the odds of service use uniquely for White adolescents. Results indicate that racial disparities characterize adolescents' mental health service use even when suicide risk increases. PMID- 17397278 TI - Suicide terrorists: are they suicidal? AB - Are suicide terrorists suicidal? A review of the worldwide literature on suicide terrorism uncovered five published empirical studies describing data collected from potential suicide terrorists or the surviving friends and families of deceased terrorists. The many discrepancies uncovered between suicide terrorists and other suicides on key factors known to underpin suicidality, suggest that such terrorists are not truly suicidal and should not be viewed as a subgroup of the general suicide population. Nonetheless, methods developed by suicidologists, such as the psychological autopsy, will help increase our understanding of the individual and group factors that underpin suicide terrorism. PMID- 17397279 TI - No-harm contracts: a review of what we know. AB - Few events in the practice of a mental health clinician can be as devastating as the suicide of a client. Although suicide is a rare occurrence, clinicians face assessment of clients' risk for harming themselves on a regular basis. One well accepted and widely practiced intervention for suicidal ideation is the use of no harm contracts (NHC), although opinions about NHCs range from enthusiasm to apathy. The existing research does not support the use of such contracts as a method for preventing suicide, nor for protecting clinicians from malpractice litigation in the event of a client suicide. PMID- 17397280 TI - Ethical, legal, and practical issues in the control and regulation of suicide promotion and assistance over the Internet. AB - The promotion of suicide and description of suicide methods on the Internet have led to widespread concern that legal control is mandated. Apart from value concerns pertaining to attitudes about suicide, the guarantee of freedom of expression presents a serious challenge to the introduction of restrictive laws. Recent developments in Australia and Europe are presented, noting jurisdictional complexity as an obstacle to effective application. Finally, scientific data of an epidemiological nature are revealed to be insufficient to warrant making causal assertions about the Internet and its relation to suicidal acts, including those of vulnerable populations. Recommendations are made with respect to public education, suicide prevention, and future research. PMID- 17397281 TI - Analysis of a national toll free suicide crisis line in South Africa. AB - The first national toll free suicide crisis line for South Africa was launched in October 2003 with the aim of providing a service dedicated to the prevention of suicide in this country. The intervention was motivated by South Africa's suicide rate which had risen higher than the global suicide rate, with the majority of attempted suicides occurring among people younger than 35 years of age (WHO, 2002). Demographic characteristics of callers were identified to evaluate the perceived helpfulness of this crisis line, so as to inform planning and implementation of future suicide prevention programs. Results showed that the majority of callers were female; between the ages of 16 and 18 years; and lived in the provinces of Gauteng, North West, or KwaZulu Natal. Callers were more likely to be from urban than rural areas; were still at school, unemployed, or studying at a tertiary institution; and had not previously attempted suicide. The majority of participants did perceive the crisis line as helpful. The continued collection of demographic data from the crisis line is recommended so that South Africa can create an updated database of areas and sectors of the population that require suicide intervention, and for planning and implementing suicide prevention programs in this country. PMID- 17397282 TI - Performance of the CES-D and its short forms in screening suicidality and hopelessness in the community. AB - A measurement scale should be short and quick to complete if it is to be practically useful. Drawing on data from a community-based survey of 2,178 people in Hong Kong, we compared five short forms (5- to 10-item) and the original version (20-item) of the Center for Epidemiologic Studies-Depression Scale (CES D; Radloff, 1977) in predicting suicidal attempts and suicidal thoughts. Short forms with as few as nine items performed in ways very similar to the full version; a version with only five items had a detectable difference from the full version. Sensitivity, specificity, and predictive values in differentiating people with and without suicidal thought or attempt change almost linearly with the cut-offs. PMID- 17397283 TI - The association of depression and sense of belonging with suicidal ideation among older adults: applicability of resiliency models. AB - Suicide among older people, especially men, is a significant problem. In this study the applicability of the compensatory, the risk-protective, the challenge, and the protective-protective models of resiliency for the prediction of suicidal ideation from depression (the risk factor) and sense of belonging to the community (the protective factor) was investigated. A total of 351 retired Australians (130 males and 221 females), with a mean age of 71.31 years (SD = 7.99), completed the Zung Depression Scale, the suicide subscale of the General Health Questionnaire, and the Sense of Belonging Instrument. When sense of belonging (psychological) was the protective factor, results indicated support for the risk-protective model for men and women, and for the compensatory model for women only. In contrast, when sense of belonging (antecedents) was the protective factor, support was evident for the compensatory model for men and women, and for the challenge model for women only. Results indicate that interventions should be developed to enhance sense of belonging among aging adults. PMID- 17397284 TI - Economic strain and suicide risk: a qualitative analysis. AB - Research at the individual level on economic strain and suicide has focused on unemployment; yet it remains unclear how unemployment and other economic strains actually affect suicide risk for the individual. In the present study Agnew's (1992, 2002), General Strain Theory was applied to a qualitative analysis of case files. Strain issues assessed include the role of goal blockage, economic loss, noxious work environments, anticipated strain, strain clusters, and vicarious strain in the generation of suicide risk. Data are from 62 cases of suicide involving economic strain from the files of an urban county medical examiner's office. The cases were classified into strain categories. Suicide risk was related to all categories of strain; however, economic strains were typically comorbid with additional strains in the genesis of suicide. Key auxiliary strains included anticipated loss of a home place, loss of a car, noxious social relationships, medical problems, death of loved ones, and involvement with the criminal justice system. Some evidence was found linking economic strain to the suicides of nonimpoverished persons. Economic strain and suicide would especially benefit by testing additional hypotheses on strain comorbidity. Unemployment combined with an anticipated eviction from one's home is recommended as a particular point of departure for future work. PMID- 17397285 TI - It's important to see the wood for the trees: a reply to Andover, Pepper, Ryabchenko, Orrico, & Gibb (2005). PMID- 17397286 TI - An historical note on suicide during the course of treatment for depression. PMID- 17397289 TI - Evaluation of agreement between two instruments in measurements of radiation dose rates in cats that underwent iodine I 131 treatment. AB - OBJECTIVE: To assess agreement between a commercially available Geiger-Meuller (GM) survey meter and millirem tissue-equivalent (TE) meter for measuring radioactivity in cats treated with sodium iodine I 131 ((131)I). ANIMALS: 15 cats with hyperthyroidism and undergoing (131)I treatment. PROCEDURES: Duplicate measurements were obtained at a distance of 30 cm from the thyroid region of each cat's neck by 2 observers who used both meters on day 3 or 5 after(131)I administration. Comparisons of measurements between meters and observers were made, with limits of agreement defined as the mean difference +/- 2 SDs of the differences. RESULTS: For observer 1, the mean of the differences in the 2 meters' measurements in all cats was 0.012 mSv/h (SD, 0.011 mSv/h). For observer 2, the mean of the differences in measurements was 0.012 mSv/h (SD, 0.010 mSv/h). For the GM meter, the mean of the differences of the 2 observers for all cats was 0.003 mSv/h (SD, 0.011 mSv/h). For the TE meter, the mean of the differences of the 2 observers for all cats was 0.003 mSv/h (SD, 0.007 mSv/h). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that there was considerable agreement between meters and observers in measurements of radioactivity in cats treated with (131)I. Measurements obtained by use of the GM meter may be approximately 0.01 mSv/h less than or 0.03 mSv/h higher than those obtained with the TE meter. If this range is acceptable for an institution's release criteria, the 2 meters should be considered interchangeable and acceptable for clinical use. PMID- 17397288 TI - Urinary biomarkers to assess exposure of cats to environmental tobacco smoke. AB - OBJECTIVE: To evaluate the use of urinary biomarkers to assess exposure of cats to environmental tobacco smoke (ETS). ANIMALS: 61 healthy client-owned cats (19 from households in which smoking was reported and 42 from households in which there was no smoking). PROCEDURES: Urine samples were obtained from each cat and assayed for total nicotine (nicotine plus nicotine glucuronide) and total cotinine (cotinine plus cotinine glucuronide) content by use of gas chromatography-mass spectrometry. In addition, total urinary content of 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a major metabolite of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, was measured by use of gas chromatography with nitrosamine-selective detection. RESULTS: Cats from households in which smoking was reported had significantly higher concentrations of total nicotine (70.4 ng/mL), total cotinine (8.53 ng/mL), and total NNAL (0.0562 pmol/mL) in urine, compared with concentrations for cats that lived in households in which there was no smoking (4.89 ng/mL, 0.74 ng/mL, and 0.0182 pmol/mL, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of these data provided biochemical evidence of exposure to ETS and uptake of tobacco-specific carcinogens by cats that live in households with smokers. Biomarkers could facilitate investigation of the health effects of ETS in cats and other species. PMID- 17397290 TI - Comparison of cardiopulmonary responses during sedation with epidural and local anesthesia for laparoscopic-assisted jejunostomy feeding tube placement with cardiopulmonary responses during general anesthesia for laparoscopic-assisted or open surgical jejunostomy feeding tube placement in healthy dogs. AB - OBJECTIVE: To evaluate the use of laparoscopic-assisted jejunostomy feeding tube (J-tube) placement in healthy dogs under sedation with epidural and local anesthesia and compare cardiopulmonary responses during this epidural anesthetic protocol with cardiopulmonary responses during general anesthesia for laparoscopic-assisted or open surgical J-tube placement. ANIMALS: 15 healthy mixed-breed dogs. PROCEDURES: Dogs were randomly assigned to receive open surgical J-tube placement under general anesthesia (n = 5 dogs; group 1), laparoscopic-assisted J-tube placement under general anesthesia (5; group 2), or laparoscopic-assisted J-tube placement under sedation with epidural and local anesthesia (5; group 3). Cardiopulmonary responses were measured at baseline (time 0), every 5 minutes during the procedure (times 5 to 30 minutes), and after the procedure (after desufflation [groups 2 and 3] or at the start of abdominal closure [group 1]). Stroke volume, cardiac index, and O(2) delivery were calculated. RESULTS: All group 3 dogs tolerated laparoscopic-assisted J-tube placement under sedation with epidural and local anesthesia. Comparison of cardiovascular parameters revealed a significantly higher cardiac index, mean arterial pressure, and O(2) delivery in group 3 dogs, compared with group 1 and 2 dogs. Minimal differences in hemodynamic parameters were found between groups undergoing laparoscopic-assisted and open surgical J-tube placement under general anesthesia (ie, groups 1 and 2); these differences were not considered to be clinically important in healthy research dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Sedation with epidural and local anesthesia provided satisfactory conditions for laparoscopic-assisted J-tube placement in healthy dogs; this anesthetic protocol caused less cardiopulmonary depression than general anesthesia and may represent a better choice for J-tube placement in critically ill patients. PMID- 17397291 TI - Effects of a magnesium adhesive cement on bone stability and healing following a metatarsal osteotomy in horses. AB - OBJECTIVE: To compare biodegradable magnesium phosphate cement (Mg-cement), calcium phosphate cement (Ca-cement), and no cement on bone repair, biocompatibility, and bone adhesive characteristics in vivo in horses. ANIMALS: 8 clinically normal adult horses. PROCEDURES: Triangular fragments (1-cm-long arms) were created by Y-shaped osteotomy of the second and fourth metatarsal bones (MTII and MTIV, respectively). Fragments were replaced in pairs to compare Mg cement (MTII, n = 8; MTIV, 8) with Ca-cement (MTIV, 8) or with no cement (MTII, 8). Clinical and radiographic evaluations were performed for 7 weeks, at which time osteotomy sites were harvested for computed tomographic measurement of bone density and callus amount, 3-point mechanical testing, and histologic evaluation of healing pattern and biodegradation. RESULTS: All horses tolerated the procedure without clinical problems. Radiographically, Mg-cement secured fragments significantly closer to parent bone, compared with Ca-cement or no treatment. Callus amount and bone remodeling and healing were significantly greater with Mg-cement, compared with Ca-cement or no cement. Biomechanical testing results and callus density among treatments were not significantly different. Significantly greater woven bone was observed adjacent to the Mg cement without foreign body reaction, compared with Ca-cement or no cement. The Mg-cement was not fully degraded and was still adhered to the fragment. CONCLUSIONS AND CLINICAL RELEVANCE: Both bone cements were biocompatible in horses, and Mg-cement may assist fracture repair by osteogenesis and fragment stabilization. Further studies are warranted on other applications and to define degradation characteristics. PMID- 17397292 TI - Assessment of cartilage degradation effects of matrix metalloproteinase-13 in equine cartilage cocultured with synoviocytes. AB - OBJECTIVE: To determine the effects of matrix metalloproteinase (MMP)-13, compared with interleukin (IL)-1alpha, on cartilage matrix molecule gene expression in a coculture system of equine cartilage explants and synoviocytes. SAMPLE POPULATION: Articular cartilage and synovium specimens harvested from femoropatellar joints of 4 horses, aged 3 to 5 years. PROCEDURES: Synoviocytes were isolated and cocultured with cartilage explants. Cultures were treated with human recombinant MMP-13 (1, 25, or 100 ng/mL) or IL-1alpha (0.01, 0.1, 1.0, or 10 ng/mL) for 96 hours, with medium exchange at 48 hours. Cartilage extracts and media were analyzed for glycosaminoglycan (GAG) content, and results were adjusted to cartilage DNA content. Quantitative PCR was performed on mRNA from cartilage (MMP-3, MMP-13, aggrecan, and collagen type IIB [COL2A1]) and synoviocytes (MMP-3 and MMP-13), and results were adjusted to 18S ribosomal subunit mRNA expression. Treatments were performed in triplicate, and the experiment was repeated 4 times. RESULTS: Cultures treated with MMP-13 or IL 1alpha had increased media GAG concentration at 48 and 96 hours. Aggrecan and COL2A1 mRNA expression were increased by application of MMP-13 or IL-1alpha. Gene expression of the catabolic mediator, MMP-3, in cartilage and synoviocytes was increased in cultures treated with MMP-13 or IL-1alpha. Expression of MMP-13 mRNA in cartilage was increased by IL-1alpha, but decreased in synoviocytes by MMP-13 treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results support the use of recombinant MMP-13 in a coculture system of synoviocytes and cartilage explants for the study of osteoarthritis. PMID- 17397293 TI - Changes in the myocardial performance index during dobutamine administration in anesthetized cats. AB - OBJECTIVE: To investigate the relationship between myocardial performance index (MPI; also known as the Tei index) and cardiac function in anesthetized cats administered dobutamine. ANIMALS: 6 adult cats. PROCEDURES: Cats were anesthetized by administration of propofol (6 mg/kg, IV), and anesthesia was maintained by administration of isoflurane. Heart rate and systolic arterial pressure (SAP) were monitored. Stroke volume, cardiac output, and aortic blood flow (ABF) were measured by use of transesophageal ultrasonography. Left ventricular fractional shortening (LVFS), mitral E-wave velocity-to-A-wave velocity (E:A) ratio, and ejection time were measured by use of transthoracic echocardiography. Dobutamine was administrated via a cephalic vein at rates of 2.5, 5.0, and 10 microg/kg/min. RESULTS: Heart rate, SAP, cardiac output, and ABF increased with dobutamine administration, whereas stroke volume significantly decreased. The LVFS significantly increased, and the E:A ratio significantly decreased. Total isovolumic time and the MPI significantly decreased. The MPI was negatively correlated (r=-0.63) with LVFS. Conversely, the MPI was positively correlated with the E:A ratio (r=0.47), stroke volume (r=0.66), and total isovolumic time (r=0.95). However, the MPI was not significantly correlated with heart rate, SAP, cardiac output, or ABF. CONCLUSION AND CLINICAL RELEVANCE: Analysis suggested that the MPI provides a sensitive clinical assessment of cardiac response to medication in cats, which may be similar to the usefulness of the MPI reported in humans. PMID- 17397294 TI - Prevalence of obese dogs in a population of dogs with cancer. AB - OBJECTIVE: To determine the body condition score (BCS) distribution for dogs examined at a teaching hospital and examine whether the BCS distribution for dogs with cancer differed significantly from the distribution for dogs without cancer. SAMPLE POPULATION: 1,777 dogs with cancer and 12,893 dogs without cancer. PROCEDURES: A retrospective prevalence case-control study was conducted that used medical records from 1999 to 2004. Information was collected on BCS (9-point system), age, breed, sex, neuter status, diagnosis, and corticosteroid administration. Body condition score at the time of examination for cancer (dogs with cancer) or first chronologic visit (dogs without cancer) was recorded. Logistic regression was used to compare BCS prevalence distributions between groups. RESULTS: The overall prevalence of obese dogs (BCS >or= 7/9) was 14.8% (2,169/14,670), and the overall prevalence of overweight dogs (BCS >or= 6/9 to < 7/9) was 21.6% (3,174/14,670). There was a significant difference in the BCS distribution between dogs with and without cancer, with a slightly lower prevalence of being overweight and obese in dogs with cancer. The prevalence of obese and overweight dogs varied with specific cancer types when compared with the prevalence for dogs without cancer. CONCLUSIONS AND CLINICAL RELEVANCE: Differences in obesity prevalence among cancer types is suggestive of an incongruous effect of this variable on cancer expression or a differential effect of specific cancer types on weight status. Systematic use of BCSs will help elucidate the association between obesity and cancer development. PMID- 17397295 TI - Phase I clinical trial of the use of zinc phthalocyanine tetrasulfonate as a photosensitizer for photodynamic therapy in dogs. AB - OBJECTIVE: To determine the threshold for acute toxicosis of parenterally administered zinc phthalocyanine tetrasulfonate (ZnPcS(4)), a candidate second generation photosensitizer, in mice and evaluate the compound's safety in a phase I clinical trial of ZnPcS(4)-based photodynamic therapy (PDT) in pet dogs with naturally occurring tumors. ANIMALS: Male Swiss-Webster mice and client-owned dogs with naturally occurring neoplasms. PROCEDURES: For the study of acute toxicosis, mice were given graded doses of ZnPcS(4). To determine safety, a rapid titration phase I clinical trial of ZnPcS(4)-based PDT in tumor-bearing dogs was conducted. RESULTS: In mice, administration of >or= 100 mg of ZnPcS(4)/kg resulted in renal tubular necrosis 24 hours after IP injection. In tumor-bearing dogs, ZnPcS(4) doses 0.05) among the groups at any time. At 6 months, the mean RC was 80.83% +/- 21.08% for group 1; the mean RRCs were 71.99% +/- 18.69% and 74.18% +/- 15.02% for groups 2 and 3, respectively. There were no statistically significant differences in RRC between groups 2 and 3. CONCLUSION: All treatments showed root coverage improvement without damage to periodontal tissues, supporting the use of CPF for treatment of root surfaces restored with RMGI or MRC as being effective over the 6-month period. PMID- 17397308 TI - Clinical evaluation of the use of locally delivered chlorhexidine in periodontal maintenance therapy. AB - BACKGROUND: The objective of this study was to evaluate clinically the effectiveness of a chlorhexidine gluconate chip in sites still showing signs of disease during periodontal maintenance therapy. METHODS: Forty-two maintenance non-smoking patients (previously treated with non-surgical scaling and root planing [SRP]), presenting at least one probing depth (PD) of 5 to 8 mm, and bleeding on probing (BOP) at single-rooted teeth were assigned randomly to two groups: treated with a chlorhexidine gluconate chip (CHIP group) and treated with SRP (SRP group). Patients were assessed for plaque index, gingival index, BOP, PD, clinical attachment level (CAL), and gingival recession at baseline, 6 weeks, and 3 and 6 months. RESULTS: Both treatments resulted in improvements in all parameters evaluated. After 6 months, a reduction in PD of 2.64 +/- 0.02 mm and 2.12 +/- 0.02 mm was observed for CHIP and SRP groups, respectively (P >0.05). The observed gain in CAL was 2.19 +/- 0.87 mm and 2.07 +/- 1.53 mm for CHIP and SRP groups, respectively (P >0.05). In deep pockets, PD reduction was 3.60 +/- 0.70 mm for CHIP group and 2.83 +/- 0.62 mm for SRP group (P = 0.01). CONCLUSIONS: Both treatments were equally effective in periodontal health reestablishment in inflamed single-root sites of maintenance patients. However, for deep pockets, the chlorhexidine gluconate chip was more effective than SRP in reducing PD. PMID- 17397309 TI - Demographic, clinical, and microbial aspects of chronic and aggressive periodontitis in Colombia: a multicenter study. AB - BACKGROUND: The microbial profile of periodontal disease varies among different human populations. This study evaluated the demographic, clinical, and microbiologic aspects of periodontitis in a multigeographic sample in Colombia. METHODS: Three hundred twenty-five patients with chronic periodontitis (CP), 158 patients with aggressive periodontitis (AgP), and 137 healthy-gingivitis controls from five regions of the country were studied. Clinical, microbial, and sociodemographic data were collected. Microbiologic identification was performed using polymerase chain reaction 16S rRNA gene on pooled subgingival samples, and the presence of Gram-negative enteric rods was evaluated by culture. Bivariate and multivariate logistic regression analyses were conducted. RESULTS: Porphyromonas gingivalis occurred in 71.5% of individuals with periodontitis, Tannerella forsythensis occurred in 58.5%, Campylobacter rectus occurred in 57.5%, Actinobacillus actinomycetemcomitans occurred in 23.6%, and enteric rods occurred in 34.5%. P. gingivalis was more common in CP and AgP than controls. A. actinomycetemcomitans was increased in AgP compared to controls and patients with CP. T. forsythensis, C. rectus, and Eikenella corrodens had a low presence in the West Pacific and Central regions, and enteric rods were increased in the Central region (P <0.05). Other sociodemographic factors were not associated with these microorganisms. CONCLUSIONS: Geographic regions do not influence the microbiota, but the microbiota may vary by geographic region. P. gingivalis, T. forsythensis, and C. rectus are the most prevalent periodontophatic microorganisms in Colombia. A. actinomycetemcomitans was more common in AgP, and a large percentage of the population studied had enteric rods in the subgingival plaque. PMID- 17397310 TI - Gingival involvement of oral lichen planus. AB - BACKGROUND: Oral lichen planus (OLP) is one of the most common oral mucosa disorders. OLP gingival involvement is very frequently observed and is characterized by a wide range of clinical appearance, symptomatology, and locations. The purpose of this study was to analyze the prevalence and clinical aspects of gingival lesions in a series of 213 patients affected by OLP. METHODS: We analyzed the prevalence and clinical aspects (location, morphology, and symptomatology) of the gingival lesions in a total of 213 clinically and histopathologically assessed patients affected by OLP. RESULTS: Gingival lesions were diagnosed in 38.4% of cases. White lesions were the most frequently observed in the gingiva (42.7%), followed by mixed lesions (31.7%), and red lesions (25.6%). For the three clinical forms, the most frequent gingival location was the simultaneous involvement of the attached and marginal gingiva. None of the white lesions presented symptomatology, whereas most red lesions or mixed forms presented symptoms that varied from discomfort to severe oral pain or a burning sensation. CONCLUSIONS: Considering the high frequency of gingival involvement in OLP, the diversity of clinical manifestations, and its potential relationship with general and oral health in patients, periodontists should be familiar with the most common clinical aspects of OLP in the gingiva. PMID- 17397311 TI - Clinical crown length of the maxillary anterior teeth preorthodontics and postorthodontics. AB - BACKGROUND: Prevalence information on excessive gingival display in postorthodontic patients is limited. By studying the size relationship of the clinical crowns of teeth, in an orthodontic population, we can begin to quantify their need for periodontal plastic surgery. METHODS: Preorthodontic and postorthodontic crown dimensions were measured on 200 plaster models, and after 5 years, 31 of the subjects were recalled to repeat the measurements in a clinical examination. The measurements included clinical crown lengths and widths of the six anterior teeth, and these values were compared to known ideals. RESULTS: This study revealed a significant increase in the length of the maxillary anterior crowns over the three examinations, yet these values were still approximately 1.5 mm shorter than ideal. The mean crown width-to-length ratio was 87% to 88% for maxillary central incisors, clearly above the accepted "ideal." In addition, 61% to 71% of maxillary central incisors exceeded allowable crown width-to-length ratios, and 61% of subjects displayed asymmetry in gingival architecture. CONCLUSIONS: Although this study only examined one aspect of excessive gingival display, to our knowledge it is the first study to show that in a predominantly young, postorthodontic population, the prevalence of non-ideal width-to-length ratios is >65% and the presence of asymmetry is >60%. Therefore, close interaction between the periodontist and the orthodontist is necessary to diagnose these conditions to provide patients with all options for improving their smile. PMID- 17397312 TI - Evaluation of the benefits of using a power toothbrush during the initial phase of periodontal therapy. AB - BACKGROUND: Studies evaluating the efficacy of oral hygiene procedures typically focus on prevention or maintenance after periodontal therapy. Little is known about the specific benefit of a power toothbrush during therapy. The objective of this study was to evaluate the clinical efficacy of power toothbrushing compared to manual toothbrushing in patients undergoing the initial phase of periodontal therapy. METHODS: This was an examiner-masked, randomized, two-arm parallel study involving 70 adults (range, 23 to 81 years) with untreated periodontitis. After an initial supragingival debridement, subjects were assigned to toothbrushing with a manual (group M) or power (group P) toothbrush. Subjects returned for evaluation after 7, 14, and 28 days. RESULTS: At day 28, the mean plaque index (PI) was significantly lower for subjects in group P than for those in group M (P = 0.006). The mean number of sites with PI >1 at days 14 and 28 and the mean number of sites with bleeding on probing at days 14 and 28 also were significantly lower in these subjects (P = 0.018 and P = 0.005, respectively, and P = 0.017 and P = 0.034, respectively). Differences in the mean gingival index, the number of sites with a gingival index >1, mean recession, mean probing depth, and the number of pockets >4 mm were not significant. CONCLUSION: Subjects using a power toothbrush during initial treatment reduced supragingival plaque to lower levels and showed significantly less bleeding on probing than subjects using a manual brush. PMID- 17397313 TI - Platelet-rich plasma: growth factors and pro- and anti-inflammatory properties. AB - BACKGROUND: Platelet-rich plasma (PRP) promotes regeneration of bone, presumably through the action of concentrated growth factors. However, it is not clear how PRP affects the inflammatory response. The purpose of this study was to analyze the growth factors in PRP and to study the effects of PRP on monocyte cytokine release and lipoxin A(4) (LXA(4)) generation. METHODS: PRP was prepared from healthy donors. Platelet-derived growth factor (PDGF)-AB, PDGF-BB, transforming growth factor-beta1, insulin-like growth factor-I, fibroblast growth factor-basic (FGF-b), epidermal growth factor (EGF), vascular endothelial growth factor, interleukin-12 (p40/70), and regulated on activation, normal T-cell expressed and secreted (RANTES) levels were evaluated by enzyme-linked immunosorbent assay and bead-based multiplexing. Peripheral blood monocytes were isolated and cultured with or without PRP. Cytokine, chemokine, and LXA(4) levels as well as monocyte chemotactic migration were analyzed. RESULTS: Growth factors were increased significantly in PRP compared to whole blood (WB) and platelet-poor plasma. Monocyte chemotactic protein-1 (MCP-1) was suppressed significantly by PRP, whereas RANTES was increased significantly in monocyte cultures. LXA(4) levels were significantly higher in PRP compared to WB. PRP stimulated monocyte chemotaxis in a dose-dependent fashion, whereas RANTES, in part, was responsible for PRP-mediated monocyte migration. CONCLUSIONS: PRP is a rich source of growth factors and promoted significant changes in monocyte-mediated proinflammatory cytokine/chemokine release. LXA(4) was increased in PRP, suggesting that PRP may suppress cytokine release, limit inflammation, and, thereby, promote tissue regeneration. PMID- 17397314 TI - Evidence of periopathogenic microorganisms in placentas of women with preeclampsia. AB - BACKGROUND: Periodontal disease is a chronic inflammatory infectious disease that may act as a focus of infection. Preeclampsia is a pregnancy-specific hypertensive disorder that often leads to maternal morbidity and mortality. Acute atherosis, the placental lesion of preeclampsia, shares many histopathological features with atherosclerosis. Recently, chronic infection was linked to the initiation of atherosclerosis. Oral pathogens have been detected in atherosclerotic plaques, where they may play a role in the development and progression of atherosclerosis. The purpose of the present study was to explore the possibility that periopathogenic bacteria may translocate into the placental tissues of women with preeclampsia. METHODS: Samples were taken from 16 placentas obtained from cesarean sections of women with preeclampsia and from 14 age matched healthy pregnant women. Polymerase chain reaction was used to detect Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum ssp., Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Treponema denticola. RESULTS: Eight of the 16 (50%) placenta specimens were positive for one or more periopathogenic bacteria in the preeclampsia group, compared to only two of the 14 samples (14.3%) from controls. Bacterial counts were statistically significantly higher in the preeclampsia group for all of the periopathogenic bacteria examined (P 6 mm (diseased). The gingival concentrations of IL-2, -4, -6, -10, -13, and -15 and interferon (IFN)-gamma were assessed by enzyme-linked immunosorbent assay. RESULTS: IL-2, -4, -6, -10, and 13 and IFN-gamma concentrations were higher within gingiva adjacent to 3 to 6 mm diseased compared to normal; IL-15 concentrations were significantly greater within normal gingiva than within diseased gingiva (P <0.001). Within gingiva adjacent to >6 mm sulci, IL-6 concentrations were significantly higher than in normal gingiva, and the concentrations of the other cytokines were similar to those in normal gingiva. Gingival IL-6 concentration correlated significantly with the adjacent gingival sulcular depth, whereas the IL-15 concentration correlated inversely with the adjacent gingival sulcular depth (P <0.001). CONCLUSIONS: IL-13, -4, and -10 had a similar concentration pattern within normal and diseased tissue; however, the concentration pattern of IL-15 was not similar to IL-2. Relatively low IL-15 concentrations within diseased gingiva suggest that IL-15 might have anti-inflammatory properties. PMID- 17397318 TI - Occurrence of periodontopathic and superinfecting bacteria in chronic and aggressive periodontitis subjects in a Colombian population. AB - BACKGROUND: Differences in the composition of subgingival microbiota in periodontitis have been observed in different geographic locations. Some of these microbial differences could have clinical significance. This study describes clinical parameters and the composition of the subgingival microbiota in chronic periodontitis (CP) and aggressive periodontitis (AgP) patients in a Colombian population. METHODS: Clinical parameters (probing depth, clinical attachment level, bleeding on probing, and plaque index) and plaque samples from 68 CP, 12 AgP, and 30 periodontally healthy subjects were analyzed. Subgingival samples were processed using culture, biochemical tests, and polymerase chain reaction for the detection of periodontal and superinfecting pathogens. The analysis of variance, chi(2), and Kruskal-Wallis tests were used to evaluate differences between groups for clinical parameters and microbiologic composition (P 0.05). Conversely, significant differences were noted in ligated teeth (with periodontitis); bone loss was directly proportional to the level of toxic compounds in the cigarettes (5.74 +/- 0.5 mm(2), 7.40 +/- 0.50 mm(2), and 6.51 +/ 0.50 mm(2) for groups 1, 2, and 3, respectively; P <0.05). CONCLUSION: Low- and high-yield cigarettes potentiated bone loss during experimental periodontitis in a directly proportional fashion. PMID- 17397323 TI - Expression of the receptor of advanced glycation end products in the gingival tissue of smokers with generalized periodontal disease and after nornicotine induction in primary gingival epithelial cells. AB - BACKGROUND: The association between smoking and periodontal disease is well established; however, the mechanism by which smoking augments the destruction of periodontal tissue is not clear. We hypothesize that smoking is related to an increased expression of receptor of advanced glycation end products (RAGE) in gingival tissues of smokers. METHODS: Gingival biopsies from five smokers and five age- and gender-matched non-smokers were examined. In addition, gingival epithelial cells (GECs) were reacted with 1 muM nornicotine for 4, 16, 24, and 48 hours for mRNA for RAGE and an additional 72 hours for protein expression. RAGE mRNA was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR), and expression of RAGE at the protein level in GECs was studied with Western blots. RESULTS: In the gingival biopsies from all 10 subjects, RT-PCR with RAGE specific primers produced a band of the predicted size. For all pairs, the smoker biopsies expressed a greater level of RAGE compared to the matched non-smokers. When viewed as groups, analysis of the band intensity indicated that RAGE mRNA in smokers was approximately 1.4-fold of the expression in non-smokers (Wilcoxon test; P = 0.031). In GECs treated with nornicotine, there was a time-dependent increase in RAGE expression up to two-fold at 48 hours. RAGE protein levels initially were reduced but increased to 1.4-fold after 48 hours. CONCLUSION: The ability of nornicotine to elevate RAGE expression in GECs, along with increased RAGE expression in inflamed gingival tissue from smokers, indicates that RAGE may be associated with periodontal disease linked to smoking. PMID- 17397324 TI - Hepatocyte growth factor levels in gingival crevicular fluid in health, disease, and after treatment. AB - BACKGROUND: Hepatocyte growth factor (HGF) is a broad-spectrum multifunctional cytokine with mitogenic, motogenic, morphogenic, and antiapoptotic functions in various types of cells. It is secreted by mesenchymal cells, including gingival fibroblasts, and its expression is induced by inflammatory cytokines, such as interleukin-1 and tumor necrosis factor-alpha, which play a potential role in periodontal destruction. Hence, the present study was carried out to assess HGF's concentration in gingival crevicular fluid (GCF) and to determine its association with periodontal disease progression. METHODS: Sixty subjects (aged 29 to 39 years) were divided into three groups based on gingival index, probing depth, clinical attachment loss, and radiographic evidence of bone loss: healthy (group I), gingivitis (group II), and chronic periodontitis (group III). The fourth group consisted of the 20 subjects in group III, from whom GCF samples were collected 6 to 8 weeks after scaling and root planing (SRP). The levels of HGF were estimated using enzyme-linked immunosorbent assay. Further, the correlation between HGF levels and clinical parameters in all groups and before and after treatment in periodontitis patients was analyzed. RESULTS: The highest mean HGF concentration was observed for group III (26.28 +/- 14.77 pg/microl), and the lowest mean concentration was observed for group I (13.99 +/- 11.24 pg/microl). Following SRP, the mean HGF concentration decreased from 26.28 +/- 14.77 pg/microl to 14.35 +/- 13.96 pg/microl, which was statistically significant. CONCLUSIONS: HGF concentration increased proportionally with the progression of periodontal disease, and HGF concentrations showed a positive correlation with the clinical parameters, suggesting that HGF plays a key role in periodontal disease progression. Also, following non-surgical periodontal therapy, the levels of HGF decrease significantly suggesting that HGF could be useful for monitoring the response to periodontal therapy. PMID- 17397325 TI - Extracellular ATP and ATPgammaS suppress the proliferation of human periodontal ligament cells by different mechanisms. AB - BACKGROUND: Periodontitis, similar to many other known inflammatory diseases, is thought to increase adenosine triphosphate (ATP) levels in extracellular spaces. Extracellular ATP acts on specific receptors to modulate the proliferation of various cell types. Platelet-rich plasma (PRP) contains high levels of ectonucleotidase activity capable of degrading ATP. The aim of this study was to investigate the effects of ATP on the proliferation of human periodontal ligament (PDL) cells and how these effects are altered by ectonucleotidases. METHODS: PDL cells were derived from healthy young volunteers. ATP content and DNA synthesis were quantified by a bioluminescence and an enzyme-linked immunosorbent assay, respectively. CD39 and p21(WAF1/cip1) expression was analyzed by Western blot. Apoptosis was evaluated by caspase-3/7 activity, terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) activity, annexin-V-binding, and DNA fragmentation. RESULTS: CD39 and ectonucleotidase-like activity were found in PDL cells and serum, respectively. Because less CD39 is expressed in freshly plated cells, both exogenous ATP and ATPgammaS, a slowly hydrolysable analog, inhibited cell proliferation under low serum conditions. ATP upregulated p21(WAF1/cip1), an inhibitor of cell-cycle progression, whereas ATPgammaS induced caspase-dependent apoptosis. Either upregulation of CD39 or added serum rescued cells from the cytostatic actions of exogenous ATP. CONCLUSIONS: In PDL cells expressing low CD39 levels, both ATP and ATPgammaS inhibited proliferation but by different mechanisms. ATP-induced growth arrest suggests that periodontal tissue regeneration is often suppressed at the site of injury. Furthermore, added ectonucleotidases protected PDL cells from ATP's cytostatic actions, suggesting that ectonucleotidase-rich PRP augments the regenerative actions of its constituent growth factors by protecting against exogenous ATP at clinical sites. PMID- 17397326 TI - Osmotic pressure and vasculature of gingiva in experimental diabetes mellitus. AB - BACKGROUND: Alterations in tissue osmotic pressure (OP) and vasculature are considered to be the inevitable aspects of an inflammatory process that subsequently alter the fluid dynamics of the tissues involved. The aim of this study was to reveal a profile of OP and vascular changes in periodontally healthy gingival tissues and analyze the relationship between them in diabetes mellitus (DM) to evaluate the possible effects of DM on the fluid dynamics of the periodontium. METHODS: Experimental DM was created by intraperitoneal streptozotocin injection in 10 periodontally healthy rats. These rats were used as the test group, and 10 systemically and periodontally healthy rats served as the control group. Gingival tissue samples obtained from the groups were used for the test procedures. OP was measured in the supernatants of these samples by a semimicrodigital osmometer. Vasculature was assessed as the alterations in vascularization (vessel number [VN]) and vasodilatation (vessel diameter [VD]) by histomorphometric means. RESULTS: There was a gross increase in the OP level of the test group (172.7 +/- 59.7 mOsm/kg) compared to the control group (11.4 +/- 4.2 mOsm/kg; P <0.001). VN was found to be significantly larger in the test group (12.7 +/- 2.8) than in the control group (6.8 +/- 1.1; P <0.001). VD was found to be smaller in the test group (10.1 +/- 2.8 microm) than in the control group (15.5 +/- 2.4 microm), and this difference was statistically significant (P <0.001). A positive correlation between OP and VN (r = 0.77; P <0.001) and a negative correlation between OP and VD (r = 0.1; P >0.05) were observed in the test group. CONCLUSION: Our results reveal that the fluid dynamics of periodontal soft tissues may be affected by the diabetic conditions in this diabetic model because of the increased OP and VN during the pathogenesis of the disease. PMID- 17397327 TI - Cementum crack formation by repeated loading in vitro. AB - BACKGROUND: In this study, the hypothesis was adopted that fatigue destruction may develop on the cementum of the root surface because of the repeated occlusal stresses loaded on the tooth as in the case of prolonged occlusal trauma. The purpose of this study is to clarify whether cemental fatigue destruction occurs when repeated stresses are loaded on the occlusal surface of a tooth. METHODS: The teeth used were five human mandible premolars from individuals 14 to 22 years of age. The teeth were freshly extracted in orthodontic treatment. They were free of decay and fillings, and their roots were straight. One half of the root was embedded in cold curing acrylic resin, and a compressive load of 5.0 kgf was repeatedly applied along the tooth axis in water 1 million times with a speed of one time per second. Changes in the tooth surface were observed by taking photographs with a stereoscopic microscope and a digital camera after every 100,000 loadings. Cracks were dyed with 2% methylene blue solution, and crack areas were measured using gray-scaled and binary-processed photographs. Data were analyzed using image analysis software. RESULTS: It was confirmed that cracks significantly developed on the buccal, mesial, and distal surfaces but not on the lingual surface; crack areas showed a tendency to significantly increase after 500,000 loadings; extension of the crack from the cemento-enamel junction to the root apex increased with time, and the average extension was 0.67 +/- 0.12 mm after 1 million loadings. CONCLUSION: This study revealed that cementum cracks developed in the cervix by repeated loadings and extended toward the root apex with time, suggesting that repeated stresses by occlusion or strong stresses such as by bruxism may trigger the development of site-specific attachment loss, which is one of the potential factors inducing periodontal diseases. PMID- 17397328 TI - Optimizing the esthetic potential of implant restorations through the use of immediate implants with immediate provisionals. AB - BACKGROUND: Few things in clinical periodontics are more challenging or potentially more rewarding than gaining an ideal result with implant-supported crowns in the esthetic zone. METHODS: This article describes a novel approach to maintaining soft tissue health and support following immediate implant placement and immediate insertion of minimally functional single-unit screw-retained fixed provisionals. RESULTS: Excellent tissue health, soft tissue profiles, and maintenance of septal bone height are documented with clinical photographs and 1 year follow-up radiographs. CONCLUSION: In addition to maximizing the chances for an optimal esthetic result, this technique also allows precise adjustment and stability of occlusal relationships for provisionals in the perioperative period. PMID- 17397329 TI - Implant-supported reconstruction in a patient with generalized aggressive periodontitis. AB - BACKGROUND: A 31-year-old female was diagnosed with generalized aggressive periodontitis based on clinical and radiographic findings. Dental implants were used to restore her dentition. The long-term success of osseointegrated implants in periodontally healthy patients has been documented well in the literature. However, only a few cases of full-mouth rehabilitation, using dental implants, have been reported in periodontally compromised patients, especially those who exhibited generalized aggressive periodontitis. METHODS: Following extraction of all teeth except #6 through #11 in the maxilla, oral rehabilitation was accomplished with the use of implant-supported restorations; detailed treatment planning and sequence are presented. RESULTS: Eight implants were loaded immediately in the mandible. Six implants were placed in the posterior maxilla with a conventional two-stage approach. The maxillary implants were loaded after a 4-month healing period. The patient was followed for 18 months postloading in the maxilla and 24 months postloading in the mandible. CONCLUSION: Successful oral rehabilitation was accomplished using osseointegrated implants in a patient with generalized aggressive periodontitis. PMID- 17397330 TI - Oral aspects of Gaucher's disease: a literature review and case report. AB - BACKGROUND: Gaucher's disease (GD) is a lysosomal storage disease with a high incidence in Ashkenazi Jews. The disease is caused by an autosomally recessive inherited deficiency of the lysosomal enzyme glucocerebrosidase, leading to the accumulation of glucocerebroside in macrophages. The lipid-laden macrophages, called Gaucher cells, can be detected in liver, spleen, and bone marrow tissues. METHODS: A case report of periodontal treatment of a 47-year-old female patient with GD with recurrent gingival hemorrhage and toothache is presented. Periapical radiographs revealed cyst-like lesions in the mandibular premolar-molar regions accompanied by severe apical root resorption, enlargement of the periodontal ligament and bone-marrow spaces, and loss of trabecular structure and radiopaque appearance of bone. An abnormally narrow and sharp coronoid process and effacement of the cortical borders of the mandibular canal were noted. RESULTS: The patient was diagnosed as having generalized severe chronic periodontitis. Treatment included oral hygiene motivation and instructions, scaling, root planing, and access flap therapy, resulting in resolution of periodontal signs and symptoms and a marked improvement in the patient's feeling of well being. CONCLUSIONS: Periodontal treatment can be effective in patients with GD. Oral findings may lead to early detection of GD, especially in the absence of clinical symptoms. Dentists should be aware of possible oral and radiographic manifestations of the disease and the role of periodontal treatment in improving patient's oral health and quality of life. PMID- 17397332 TI - Legislation addresses critical shortage of veterinarians in public health. PMID- 17397333 TI - Sees specialized education as limiting future needs. PMID- 17397334 TI - What is your diagnosis? Pulmonary abscess. PMID- 17397335 TI - What is your diagnosis? Mycoplasma hemofelis infection. PMID- 17397336 TI - Diagnostic imaging in veterinary dental practice. Periapical cyst. PMID- 17397337 TI - ECG of the month. Sinus arrhythmia with first-degree atrioventricular block. PMID- 17397338 TI - Sexual harassment issues in veterinary practice. PMID- 17397339 TI - Day-to-day variability of blood glucose concentration curves generated at home in cats with diabetes mellitus. AB - OBJECTIVE: To evaluate day-to-day variability in blood glucose curves (BGCs) generated at home and at the clinic for cats with diabetes mellitus. DESIGN: Prospective study. ANIMALS: 7 cats with diabetes mellitus. Procedures-BGCs generated at home on 2 consecutive days and within 1 week at the clinic were obtained twice. On each occasion, insulin dose, amount of food, and type of food were consistent for all 3 BGCs. Results of curves generated at home were compared with each other and with the corresponding clinic curve. RESULTS: Differences between blood glucose concentration determined after food was withheld (fasting), nadir concentration, time to nadir concentration, maximum concentration, and mean concentration during 12 hours had high coefficients of variation, as did the difference between fasting blood glucose and nadir concentrations and area under the curve of home curves. Differences between home curve variables were not smaller than those between home and clinic curves, indicating large day-to-day variability in both home and clinic curves. Evaluation of the paired home curves led to the same theoretical recommendation for adjustment of insulin dose on 6 of 14 occasions, and evaluation of home and clinic curves resulted in the same recommendation on 14 of 28 occasions. Four of the 6 paired home curves in cats with good glycemic control and 2 of the 8 paired home curves in cats with poor glycemic control led to the same recommendation. CONCLUSIONS AND CLINICAL RELEVANCE: Considerable day-to-day variability was detected in BGCs generated at home. Cats with good glycemic control may have more reproducible curves generated during blood collection at home than cats with poorer control. PMID- 17397341 TI - Treatment of thoracolumbar spinal cord compression associated with Histoplasma capsulatum infection in a cat. AB - CASE DESCRIPTION: A 7-year-old domestic shorthair cat with a 2-month history of decreased appetite and weight loss was examined because of paraparesis of 1 week's duration that had progressed to paraplegia 3 days earlier. CLINICAL FINDINGS: Neurologic examination revealed normo- to hyperreflexia and absence of deep pain sensation in the hind limbs and thoracolumbar spinal hyperesthesia. Neuro-anatomically, the lesion was located within the T3 through L3 spinal cord segments. Biochemical analysis and cytologic examination of CSF revealed no abnormalities. Radiography revealed narrowing of the T11-12 intervertebral disk space and intervertebral foramen suggestive of intervertebral disk disease. Myelography revealed an extradural mass centered at the T12-13 intervertebral disk space with extension over the dorsal surfaces of T11-13 and L1 vertebral bodies. TREATMENT AND OUTCOME: A right-sided hemilaminectomy was performed over the T11-12, T12-13, and T13-L1 intervertebral disk spaces, and a space-occupying mass was revealed. Aerobic bacterial culture of samples of the mass yielded growth of a yeast organism after a 10-day incubation period; histologically, Histoplasma capsulatum was identified. Treatment with itraconazole was initiated. Nineteen days after surgery, superficial pain sensation and voluntary motor function were evident in both hind limbs. After approximately 3.5 months, the cat was ambulatory with sling assistance and had regained some ability to urinate voluntarily. CLINICAL RELEVANCE: In cats with myelopathies that have no overt evidence of fungal dissemination, differential diagnoses should include CNS histoplasmosis. Although prognosis associated with fungal infections of the CNS is generally guarded, treatment is warranted and may have a positive outcome. PMID- 17397342 TI - Immune-mediated erythroid and megakaryocytic aplasia in a cat. AB - CASE DESCRIPTION: A 6-month-old domestic shorthair cat was evaluated because of acute lethargy. CLINICAL FINDINGS: Severe nonregenerative anemia and thrombocytopenia were identified. Cytologic examination of a bone marrow aspirate revealed selective erythroid and mega-karyocytic aplasia and a high number of apparently normal small lymphocytes. Infectious agents implicated in feline hematologic disorders were excluded on the basis of serologic tests or PCR amplification, including FeLV, Ehrlichia canis, Anaplasma phagocytophilum, Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum, and Candidatus Myco plasma turicensis. TREATMENT AND OUTCOME: A 10-day course of prednisolone administration did not improve the hematologic disorder. Administration of human polyclonal immunoglobulins preceded increased reticulocyte count by 3 days. A second bone marrow examination confirmed restoration of erythroblasts and megakaryocytes. After 1 relapse, the disease was successfully controlled with prednisolone for > 3 years. CLINICAL RELEVANCE: Immune-mediated bone marrow aplasia is rare in cats and usually affects only erythrocyte progenitors. Concomitant involvement of erythroid and megakaryocytic cell lines can be successfully treated via immunosuppressive therapy. Human immunoglobulins seem to be well tolerated in cats; however, proof of a beneficial effect requires further study. PMID- 17397343 TI - Immune-mediated hemolytic anemia in an eclectus parrot. AB - CASE DESCRIPTION: A 2-year-old female Solomon Island eclectus parrot (Eclectus roratus) was evaluated by a veterinarian because of a 4-day history of progressive lethargy, weakness, poor appetite, and inactivity. The bird was referred to a veterinary teaching hospital for further examination. CLINICAL FINDINGS: Clinicopathologic analyses revealed that the parrot had marked regenerative anemia, autoagglutination, and biliverdinuria. Small, rounded RBCs (thought to be spherocytes) were detected in blood smears. The abnormal findings met the diagnostic criteria for dogs with primary immune-mediated hemolytic anemia. However, analyses of blood samples for lead and zinc concentrations and plasma bile acids concentrations; the use of PCR assays for Chlamydophila psittaci, psittacine circovirus 1 (causative agent of beak and feather disease), and polyomavirus; and microbial culture and Gram staining of feces did not reveal a cause for the hemolytic anemia. TREATMENT AND OUTCOME: Although administration of immunosuppressive doses of cyclosporine was initiated, there was a rapid progression of disease, which lead to death of the parrot before this treatment could be continued long-term. Lack of an identifiable underlying disease (confirmed by complete histologic examinations at necropsy) supported the diagnosis of primary immune-mediated hemolytic anemia. CLINICAL RELEVANCE: Primary immune-mediated hemolytic anemia has not been widely reported in psittacine birds. A comprehensive evaluation and complete histologic examination of tissues to rule out underlying disease processes are required to definitively establish a diagnosis of primary immune-mediated hemolytic anemia in parrots. Primary immune-me-diated hemolytic anemia should be considered as a differential diagnosis for regenerative anemia in a parrot. PMID- 17397344 TI - Chronic nasal discharge in cats: 75 cases (1993-2004). AB - OBJECTIVE: To identify the most common etiologic diagnosis and any historical, physical, or other diagnostic variables associated with a definitive etiologic diagnosis for chronic nasal discharge in cats. Design-Retrospective case series. ANIMALS: 75 cats with nasal discharge of >/= 1 month's duration. PROCEDURES: Medical records of affected cats were reviewed for information on signalment, clinical signs, duration and type of nasal discharge, results of clinical examination, laboratory findings, and advanced imaging findings. RESULTS: A specific etiologic diagnosis for nasal discharge was identified in only 36% of cats. Neoplasia (carcinoma or lymphoma) was the most common etiologic diagnosis. Character and location of nasal discharge did not contribute greatly toward a specific etiologic diagnosis. Sneezing and vomiting were the most common concurrent clinical signs. Routine CBC, serum biochemical panel, and urinalysis did not contribute to a specific etiologic diagnosis. An etiologic diagnosis was more likely in older cats and cats that underwent advanced imaging studies and nasal biopsy. CONCLUSIONS AND CLINICAL RELEVANCE: Although advanced diagnostic testing, including imaging studies and biopsy, increases the likelihood of achieving an etiologic diagnosis, the cause of chronic nasal discharge in cats often remains elusive. PMID- 17397345 TI - Prevalence of Neospora caninum and persistent infection with bovine viral diarrhea virus in dairy-breed steers in a feedlot. AB - OBJECTIVE: To determine the prevalence and effect of Neospora caninum infection and persistent infection (PI) with bovine viral diarrhea virus (BVDV) on weight gain, morbidity, and mortality rate in dairy-breed steer calves located on a feedlot in California. DESIGN: Prospective cohort observational study. ANIMALS: 900 dairy-breed steer calves in 2 pens. PROCEDURES: The 3- to 4-month-old calves were evaluated for serum antibodies against N caninum and infection with BVDV at entry to the feedlot. Five months later, sera were again analyzed for anti-N caninum antibodies; calves that were determined to have BVDV infection initially were retested to evaluate PI status. Average daily gain, morbidity, and deaths were recorded for all calves. RESULTS: Among 900 calves, prevalence of N caninum infection was 16.7% (95% confidence interval, 14.3% to 19.3%); prevalence of BVDV associated PI was 0.2% (95% confidence interval, 0.03% to 0.9%). Morbidity rate and time to first illness were not significantly different between calves that were seropositive or seronegative for N caninum. At the second sample collection, weight and average daily gain of calves that were seropositive for N caninum was less than that of seronegative steers in 1 pen, whereas these measures did not differ between groups in the other pen. Statistical power was insufficient to evaluate the effect of BVDV PI on any outcome measurement. CONCLUSIONS AND CLINICAL RELEVANCE: Although N caninum serostatus had no significant effect on morbidity rate, some seropositive calves had reduced growth, compared with seronegative calves, 5 months after entry to the feedlot. PMID- 17397347 TI - Use of a circular external skeletal fixator for stabilization of a comminuted diaphyseal metatarsal fracture in an alpaca. AB - CASE DESCRIPTION: A 3-year-old male alpaca was evaluated because of non-weight bearing lameness (grade 5/5) in the left hind limb. CLINICAL FINDINGS: Clinical and radiographic examination revealed a closed, comminuted, nonarticular, displaced diaphyseal fracture of the left third and fourth metatarsal bones. TREATMENT AND OUTCOME: Initial attempts at treatment via reduction of the fracture under traction and subsequent application of a cast were unsuccessful, and more stable fracture fixation was pursued. The alpaca underwent closed reduction of the fracture, which was stabilized by the application of a 3-ring circular external skeletal fixator (CESF). Improved weight bearing on the affected limb was evident soon after surgery and gradually increased; full weight bearing was evident by the seventh day after discharge from the hospital (day 20 after application of the CESF). Lameness was hardly noticeable during walking at that time. After 3 months, complete fracture healing was evident and the CESF was removed; mild outward rotation of the distal fragment and metatarsophalangeal joint was present. A Robert Jones bandage was applied to the limb, and the alpaca was kept in a stall for another 4 weeks. Eleven months after CESF application, the owners and referring veterinarian reported that the alpaca was healthy, not lame, and serving as a stallion without apparent impediment. CLINICAL RELEVANCE: Although mostly restricted to small animals, application of a CESF can be a viable alternative for management of long bone fractures in South American camelids. PMID- 17397348 TI - Coelioscopic-assisted prefemoral oophorectomy in chelonians. AB - OBJECTIVE: To describe a coelioscopic-assisted prefemoral oophorectomy technique for use in chelonians. DESIGN: Descriptive report. ANIMALS: 11 adult female turtles (6 red-eared sliders, 2 box turtles, 1 painted turtle, 1 four-eyed turtle, and 1 Chinese red-necked pond turtle). Five turtles required oophorectomy because of reproductive tract disease; the remaining 6 underwent elective oophorectomy. PROCEDURES: Turtles were anesthetized and positioned in dorsal recumbency. An incision was made in the prefemoral fossa, and a 2.7-mm rigid endoscope was inserted into the coelomic cavity and used to identify the ovaries. Each ovary was grasped with forceps and exteriorized through the prefemoral incision. The ovarian vasculature was ligated, and the mesovarium was transected. Closure was routine. RESULTS: In 8 turtles, bilateral oophorectomy was performed through a single incision. In 2 turtles, unilateral oophorectomy was performed in an attempt to maintain reproductive potential. In 1 turtle with a unilateral ovarian remnant from a previous surgery, unilateral oophorectomy was performed. Nine turtles recovered. One box turtle with severe hepatic lipidosis died 7 days after surgery. A second box turtle died 2 days after removal of retained eggs and a large bacterial granuloma. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that coelioscopic-assisted prefemoral oophorectomy is a practical and safe method for treating reproductive disorders and performing elective oophorectomy in turtles. This technique represents a potential alternative to plastron osteotomy in sexually mature chelonians. PMID- 17397349 TI - Mucocutaneous manifestations of disseminated histoplasmosis in patients with acquired immunodeficiency syndrome: particular aspects in a Latin-American population. AB - BACKGROUND: Mucocutaneous lesions in human immunodeficiency virus (HIV)-infected patients with disseminated histoplasmosis have a wide spectrum of clinical manifestations, making its diagnosis difficult. Studies have been restricted to case reports and series with small numbers of patients not specifically focusing on the dermatological aspects of histoplasmosis. AIMS: To describe the characteristics of mucocutaneous lesions of disseminated histoplasmosis in HIV infected patients. METHODS: A retrospective and prospective study was conducted on 36 HIV-infected patients with mucocutaneous histoplasmosis in a tertiary-care hospital in Brazil. RESULTS: Mucocutaneous histoplasmosis was diagnosed by histopathology in 33 of the 36 patients (91%) and/or culture in 23 (64%). Their CD4+ cell counts ranged from 2 to 103 cells/mm(3). The average number of different morphological types of lesions was three per patient. Despite the variability of the lesions, papules (50%), crusted papules (64%) and oral mucosal erosions and/or ulcers (58%) were the most frequent dermatological lesions. A diffuse pattern of distribution of the skin lesions was found in 58% of the cases. There was significant association between the CD4+ cell counts and the morphological variability of lesions per patient. Variation in the lesions seemed to be associated with higher CD4+ cell counts. CONCLUSION: Doctors caring for HIV infected patients should be aware of the wide spectrum of dermatological lesions observed in disseminated histoplasmosis and the importance of detecting and isolating the fungus in mucocutaneous tissues. PMID- 17397350 TI - Cutaneous melanoma: a population-based epidemiology report with 989 patients in Hong Kong. AB - Studies in white populations have confirmed advanced age as a risk factor for cutaneous melanoma, but in nonwhite populations, its role is less clear. To clarify a possible association in our local population, comprising 94.9% Chinese, a retrospective epidemiological study of 20 years of data on cutaneous melanoma between 1983 and 2002 from a central cancer registry in Hong Kong was conducted. There were 989 new cases and 378 death cases registered, and analysis showed that both the incidence and mortality rate of cutaneous melanoma increase with increasing age. Advanced age is thus confirmed as a risk factor for cutaneous melanoma in our local population. In an ageing population, the estimated future incidence and mortality rate of cutaneous melanoma are likely to increase. PMID- 17397351 TI - Treatment of severe scleroderma skin ulcers with recombinant human erythropoietin. AB - Systemic sclerosis (SSc) is frequently complicated by skin ulcers, often unresponsive to traditional treatments. A preliminary evaluation of the effects of recombinant human erythropoietin (rHuEPO) was carried out in 14 patients with SSc with nonhealing, severe cutaneous ulcers. Patients received rHuEPO subcutaneously at a dosage of 150 IU/kg 3 times weekly for 2 weeks, twice weekly for the next 2 weeks, and then once weekly for 1 month. At follow-up 3-6 months from the beginning of the treatment, six patients showed complete resolution of the skin ulcers, while a significant reduction (> 60%) in lesional areas was obtained in the other eight patients (mean +/- SD ulcer area reduced from 27.6 +/ 28 to 5.3 +/- 7.8 cm(2); P<0.005). Moreover, patients' quality of life significantly improved (pain, as measured on visual analogue scale reduced from 96 +/- 5 to 46 +/- 17 points; P=0.0001; disability as measured by the Health Assessment Questionnaire-Disability Index reduced from 1.6 +/- 0.5 to 0.9 +/- 0.4 points; P=0.0001). The rHuEPO may represent a novel treatment of nonhealing scleroderma skin ulcers, suggesting some important aetiopathological implications. PMID- 17397352 TI - Bullous pemphigoid associated with dermatomyositis and colon carcinoma. AB - It has been suggested that both bullous pemphigoid (BP) and dermatomyositis are associated with internal malignancies. However, there are no reports of the coexistence of these three conditions in one patient. We report the first case, to our knowledge. of coexistent BP, dermatomyositis and colon carcinoma. An 81 year-old Japanese woman was diagnosed with dermatomyositis based on muscular weakness and tenderness of the extremities, increased levels of circulating muscle enzymes and histological inflammatory changes in skeletal muscle and skin. Carcinoma of the sigmoid colon was detected during screening for internal malignancy; she was therefore surgically treated. Ten days after the operation, several large bullous eruptions appeared on her extremities. From the histopathological, immunofluorescence microscopy and BP180 ELISA study findings, we diagnosed the bullous eruptions as BP. Coexistence of BP with dermatomyositis or colon carcinoma is rare. Although the reason why our case exhibited these three disorders is not understood, it is suggested that these conditions may be associated with epitope spreading. PMID- 17397353 TI - Efficacy of erythromycin for psoriasis vulgaris. AB - Psoriasis is characterised by the presence of neutrophil overactivation and overproduction of interleukin (IL)-6 and IL-8 from keratinocytes. It is now clear that macrolide antibiotics have anti-inflammatory effects, such as inhibition of IL-6, IL-8 and tumour necrosis factor-alpha, perhaps by suppressing the transcription factor nuclear factor-kappaB or activator protein-1, and reduction of neutrophil activity. It is thus possible that macrolides might be a candidate for adjunctive treatment of psoriasis. In this study, we investigated the effectiveness of treatment with the macrolide antibiotic, erythromycin, for skin lesions and pruritus of patients with psoriasis. In total, 60 patients with psoriasis, especially pruritic psoriasis, were included. This was an open-label study and the analysis was on an intention-to-treat basis. Oral macrolide antibiotics and topical corticosteroids were given to the study group of 36 patients. The control group (24 patients) were treated only with topical corticosteroids. After a 4-week treatment period, scores on the Psoriasis Area and Severity Index (PASI) at baseline and at the end of the treatment, and the effectiveness in reducing itching were compared within and between both groups. Although there was no statistically significant difference between the baseline mean PASI of the two groups (P=0.81), there was a statistically significant difference between the mean PASI of the two groups at the end of the treatment (P=0.023, 95% confidence interval: - 3.45 to - 0.27). The comparison of the mean difference in PASI yielded a statistically significant difference (P=0.03, 95% confidence interval 0.73-3.55). Our study suggests that macrolides could be used as one of the adjunctive therapies for psoriasis vulgaris. PMID- 17397354 TI - Tumour-like muscular sarcoidosis. AB - Sarcoidosis is a multisystem granulomatous disease of unknown aetiology with variable manifestations, which may affect virtually any organ. Muscular sarcoidosis is a rare entity, and among this group of muscular lesions, the tumour-like muscular sarcoidosis subtype is extremely rare. We report on two sarcoidosis cases that presented muscular sarcoid lesions with subcutaneous tumours. PMID- 17397355 TI - Levamisole can reduce the high serum tumour necrosis factor-alpha level to a normal level in patients with erosive oral lichen planus. AB - In this study, we measured the baseline serum levels of tumour necrosis factor (TNF)-alpha in 158 patients with oral lichen planus (OLP) and in 54 normal control subjects. In total, 60 patients with erosive OLP (EOLP) with relatively high TNF-alpha levels were treated with levamisole and the serum TNF-alpha levels measured after treatment. We found that the mean +/- SD serum level of TNF-alpha in patients with either type of EOLP (12.0 +/- 1.7 pg/mL, P<0.005), major EOLP (15.5 +/- 4.4 pg/mL, P<0.001), minor EOLP (11.1 +/- 1.8 pg/mL, P<0.01), or nonerosive OLP (6.1 +/- 1.7 pg/mL, P<0.05) was significantly higher than that (3.8 +/- 0.2 pg/mL) of normal control subjects. Treatment with levamisole for a period of 0.5-7.5 months significantly reduced the mean serum TNF-alpha level from 22.6 +/- 3.4 pg/mL to 6.2 +/- 0.8 pg/mL (P<0.001) in 60 patients with EOLP. We conclude that levamisole can reduce high serum TNF-alpha levels to normal in patients with EOLP. PMID- 17397356 TI - Psychocutaneous syndromes: a call for revised nomenclature. AB - We propose abandoning three outdated, pejorative diagnostic terms: 'trichotillomania', 'delusions of parasitosis', and 'neurotic excoriation'. These insensitive terms are offensive to patients and become a barrier to treatment. We suggest the following alternative patient-centred nomenclature: 'neuromechanical alopecia', 'pseudoparasitic dysaesthesia', and (simply) 'excoriation'. We briefly review clinical and historical aspects of these disorders. PMID- 17397357 TI - Collagen naevus in a zosteriform distribution. PMID- 17397358 TI - Fibrillar pattern of acral melanocytic naevi may be a dermatoscopic, but not a histological artefact. PMID- 17397359 TI - Extensive mammary Paget's disease. PMID- 17397360 TI - A case of porphyria cutanea tarda resulting in digital amputation and improved by anastrazole. PMID- 17397361 TI - Does prior treatment with fumaric acid esters predispose to tuberculosis in patients on etanercept? PMID- 17397362 TI - Lack of response of severe steroid-dependent chronic urticaria to rituximab. PMID- 17397363 TI - Secondary cryofibrinogenaemia due to bronchial adenocarcinoma presenting as cutaneous necrosis. PMID- 17397364 TI - Trichophyton tonsurans infection mimicking tinea imbricata. PMID- 17397365 TI - Physical health, illicit drug use, and demographic characteristics in rural stimulant users. AB - CONTEXT: There is growing concern about illicit rural stimulant use, especially regarding methamphetamine use and its health consequences. PURPOSE: The present study describes associations between aspects of stimulant use and illness experience in rural areas, with additional focus on the role of demographic characteristics in these associations. METHODS: The research participants were 710 stimulant drug users who were recruited from rural areas of Arkansas, Kentucky, and Ohio using Heckathorn's respondent-driven sampling method. Health was measured by self-reports of perceived health and extent of current, recent, and lifelong health problems. Drug use was measured with self-reports of type and frequency of use. FINDINGS: Several associations were found between drug use and illness, controlling for demographics. Stimulant use pattern related significantly with the sum of health problems in the previous 6 months and the sum of lifetime illness diagnoses, after adjustment for demographic factors. Extent of illicit drug use in the past month and self-perceived drug and alcohol problems were associated with several measures of health. CONCLUSIONS: In this sample of stimulant users, methamphetamine use was associated with fewer recent medical problems than crack cocaine, combined crack and powder cocaine use, and use of all 3 of these stimulants. These results, across the 3 sites, suggest that prevalent assumptions about the methamphetamine "plague" and its negative health consequences must be viewed cautiously and examined with additional research. PMID- 17397366 TI - Use of critical access hospital emergency rooms by patients with mental health symptoms. AB - CONTEXT: National data demonstrate that mental health (MH) visits to the emergency room (ER) comprise a small, but not inconsequential, proportion of all visits; however, we lack a rural picture of this issue. PURPOSE: This study investigates the use of critical access hospital (CAH) ERs by patients with MH problems to understand the role these facilities play in rural MH needs and the challenges they face. METHODS: Primary data were collected through the combination of a telephone survey and ER visit logs. Our sampling frame was the universe of CAHs at the time the survey was fielded. KEY FINDINGS: About 43% of CAHs surveyed operate in communities with no MH services, while 9.4% of all logged visits were by patients identified as having some type of MH problem. The most common problems identified were affective disorders, substance abuse, anxiety, and psychotic disorders. Only 32% of CAHs have access to on-site detoxification and 2% have inpatient psychiatric services, meaning that patients in need of these services typically must leave their communities to gain treatment. CONCLUSIONS: The lack of community resources may impact CAHs' ability to assist patients with MH problems. Among those with a primary MH condition, 21% left the ER with no or unknown treatment, as did 51% of patients whose MH condition was secondary to their emergent problem. Patients in need of detoxification or inpatient psychiatric services often must travel over an hour to obtain these services, potentially creating significant issues for themselves and their families. PMID- 17397367 TI - Intensive care in critical access hospitals. AB - CONTEXT: Although critical access hospitals (CAHs) have limitations on number of acute care beds and average length of stay, some of them provide intensive care unit (ICU) services. PURPOSE: To describe the facilities, equipment, and staffing used by CAHs for intensive care, the types of patients receiving ICU care, and the perceived impact of closing the ICU on CAH staff and the local community. METHODS: A semistructured interview of directors of nursing at CAHs that provide intensive care services. RESULTS: Two thirds of CAHs that provide intensive care do so in a distinct unit. Most have continuous or computerized electrocardiography and ventilators. Other ICU equipment common in larger hospitals was reported less frequently. Nurse:patient ratio ranged from 1:1 to 1:3, and some or all nursing staff have advanced cardiac life support certification. Most CAHs admit patients to the ICU daily or weekly, primarily treating cardiac, respiratory, gastrointestinal, endocrine, and drug- or alcohol related conditions. ICUs are also used for postsurgical recovery. Respondents felt that closure of the ICU would be burdensome to patients and families, result in lost revenue, negatively impact staff, and affect the community's perception of the hospital. CONCLUSIONS: Intensive care services provided by CAHs fall along a continuum, ranging from care in a unit that resembles a scaled-down version of ICUs in larger hospitals to care in closely monitored medical-surgical beds. Nurse to patient ratio, not technology, is arguably the defining characteristic of intensive care in CAHs. Respondents believe these services to be important to the well-being of the hospital and of the community. PMID- 17397368 TI - The health trade-off of rural residence for impaired older adults: longer life, more impairment. AB - CONTEXT: Years lived with and without physical impairment are central measures of public health. PURPOSE: We sought to determine whether these measures differed between rural and urban residents who were impaired at the time of a baseline measurement. We examined 16 subgroups defined by rural/urban residence, gender, race, and education. METHODS: This is a 20-year retrospective cohort study, following 2,939 Americans who were aged 65-69 in 1982 and physically impaired at the time of the baseline measurement, with data from the National Long-Term Care Survey. Interpolated Markov chain analysis and microsimulation estimated life expectancy at age 65 and expected number of years with physical impairment. Impairment was defined as requiring help in 1 or more activities of daily living. FINDINGS: Among older individuals with physical impairments at baseline, rural residents lived notably longer than urban residents. In all but 1 group, rural residents lived more years with physical impairment, and they also had a notably larger proportion of remaining life impaired. CONCLUSIONS: Results suggest a notable public health impact of rural residence for impaired individuals, a longer expected period of impairment. Needs for services for people with impairments may be greater in rural areas. PMID- 17397369 TI - Predicting rural practice using different definitions to classify medical school applicants as having a rural upbringing. AB - PURPOSE: This study determines the relationship between a medical school applicant's rural background and the likelihood of rural practice using different definitions of rural background. METHODS: Cohort study of 599 physicians who entered the University of Virginia School of Medicine in 1990-1995 and graduated in 1994-1999. The "rurality" of the applicants' backgrounds was assessed by coding applicants' high school, college, and permanent addresses using 4 definitions of rural. In addition, most physicians responded to a survey question "Did you grow up in a rural area?" The same 4 definitions of rural were used to assess the rurality of practice locations. Logistic regression models were used to predict the simultaneous effect of different definitions of rural background, gender, and applicants' career preference at matriculation on the probability of practicing in rural areas. FINDINGS: In univariate analyses, the high school, college, and permanent addresses were all predictive of rural practice using 1 or more definitions of rural. In the multivariate analysis, only the "grew up rural" self-description was predictive of rural practice location using 3 of the definitions of rural. In a secondary analysis, the grew up rural self-description and applicants' career preferences at matriculation were predictors of rural practice. CONCLUSIONS: Readily available addresses were predictive of rural practice. In a multivariate analysis, physicians' self-description about having grown up in a rural area was the best predictor of rural practice. Recruiting more applicants who match this definition of rural background should increase the number of rural physicians. PMID- 17397370 TI - Recruitment of physicians to rural America: a view through the lens of Transaction Cost Theory. AB - CONTEXT: Many rural hospitals in the United States continue to have difficulties recruiting physicians. While several studies have examined some of the factors affecting the nature of this problem, we know far less about the role of economic incentives between rural providers and physicians. PURPOSE: This conceptual article describes an economic theory of organization called Transaction Cost Theory (TCT) and applies it to rural hospital-physician relationships to highlight how transaction costs affect the type of contractual arrangement used by rural hospitals when recruiting physicians. METHODS: The literature is reviewed to introduce TCT, describe current trends in hospital contracting with physicians, and develop a TCT contracting model for analysis of rural hospital physician recruitment. FINDINGS: The TCT model predicts that hospitals tend to favor contractual arrangements in which physicians are full-time employees if investments in physical or other assets made by hospitals cannot be easily redeployed for other services in the health care system. Transaction costs related to motivation and coordination of physician services are the key factors in understanding the unique contractual difficulties faced by rural providers. CONCLUSIONS: The TCT model can be used by rural hospital administrators to assess economic incentives for physician recruitment. PMID- 17397371 TI - Rural/urban differences in barriers to and burden of care for children with special health care needs. AB - PURPOSE: To examine the barriers and difficulties experienced by rural families of children with special health care needs (CSHCN) in caring for their children. METHODS: The National Survey of Children with Special Health Care Needs was used to examine rural-urban differences in types of providers used, reasons CSHCN had unmet health care needs, insurance and financial difficulties encountered, and the family burden of providing the child's medical care. We present both unadjusted and adjusted results to allow consideration of the causes of rural urban differences. FINDINGS: Rural CSHCN are less likely to be seen by a pediatrician than urban children. They are more likely to have unmet health care needs due to transportation difficulties or because care was not available in the area; there were minimal other differences in barriers to care. Families of rural CSHCN are more likely to report financial difficulties associated with their children's medical needs and more likely to provide care at home for their children. CONCLUSIONS: Examining results from both unadjusted and adjusted odds ratios shows that the burden of care for families of rural CSHCN stems both from socioeconomic differences and health system differences. Policies aimed at achieving equity for rural children will require focusing on both individual factors and the health care infrastructure, including increasing insurance coverage to lessen financial difficulties and addressing the availability of providers in rural areas. PMID- 17397372 TI - Fatal passenger vehicle crashes with at least 1 driver younger than 15 years: a fatality analysis reporting system study. AB - CONTEXT: A small number of fatalities continue to occur due to motor vehicle crashes on highways in which at least 1 passenger vehicle (automobile, van, or small truck) is driven by a child younger than 15 years. PURPOSE: The purpose of this study was to extend previous work suggesting that such crashes occur frequently in the Southern states and have relatively high rates in rural areas in the South and Great Plains. METHODS: This study utilizes data for the 5-year period 1999-2003 from the National Highway Transportation Safety Administration's online Fatality Analysis Reporting System. All cases were identified in which at least 1 conventional passenger vehicle in a fatal crash was being driven by a child younger than 15 years. FINDINGS: During the 5-year period, 350 fatal crashes occurred with at least 1 driver younger than 15 years involved. Twenty one of these drivers were licensed (11) or driving with a learner's permit (10). A total of 987 individuals in 419 vehicles were involved in these crashes, and 402 deaths resulted (1.16 deaths/crash). These crashes occurred primarily in Texas, Florida, Arkansas, and Arizona, but the highest rates per 100,000 children were found in North and South Dakota and predominantly in a band of Intermountain and Plains states. There was a strong correlation between crash rates and several measures of rurality. CONCLUSIONS: Crashes involving young, largely unlicensed, drivers account for about 70 deaths yearly. PMID- 17397373 TI - Rural versus suburban primary care needs, utilization, and satisfaction with telepsychiatric consultation. AB - CONTEXT AND PURPOSE: Rural and suburban populations remain underserved in terms of psychiatric services but have not been compared directly in terms of using telepsychiatry. METHODS: Patient demographics, reasons for consultation, diagnosis, and alternatives to telepsychiatric consultation were collected for 200 consecutive, first-time telepsychiatric consultations at rural and suburban clinics. FINDINGS: Rural patients were more likely than suburban patients to be younger than 18 years, using Medicaid, and needing treatment planning (lest they be referred out of the community). Rural patient and primary care physician satisfaction was higher than that of suburban counterparts. CONCLUSION: Telepsychiatry programs may enhance access, satisfaction, and quality of rural care. PMID- 17397374 TI - Information seeking and intentions to have genetic testing for hereditary cancers in rural and Appalachian Kentuckians. AB - CONTEXT: Research is limited regarding the potential of genetic testing for cancer risk in rural Appalachia. PURPOSE: This study examined perceptions of genetic testing in a population sample of Kentuckians, with a focus on Appalachian and rural differences. The goals were to examine cultural and psychosocial factors that may predict intentions to test for hereditary cancer, need for help with information seeking for decision making about genetic testing for hereditary cancer, and amount of help needed with information seeking for decision making about genetic testing for hereditary cancer in this population. METHODS: Analysis of data from a general social survey of adults using random digit dialing in Kentucky (N = 882). FINDINGS: An ordinal regression found that younger age, having a family history of cancer, and greater worry predicted greater intentions to seek genetic testing. A logistic regression found that having more education, excellent subjective knowledge of genetics, and less worry about cancer predicted less need for help in seeking information about testing. An ordinal regression found that less subjective knowledge of genetics and greater worry predicted greater amount of help needed. CONCLUSIONS: Additional counseling to explain limitations of genetic testing may be needed. Further, those with less knowledge about genetics and more worry about hereditary cancer may have greater need for help with information seeking for decision making, a need that may be further exacerbated by the lack of medical professionals, particularly genetic counselors, who may provide information about genetic testing in rural, Appalachian Kentucky. PMID- 17397375 TI - Prioritizing threats to patient safety in rural primary care. AB - CONTEXT: Rural primary care is a complex environment in which multiple patient safety challenges can arise. To make progress in improving safety with limited resources, each practice needs to identify those safety problems that pose the greatest threat to patients and focus efforts on these. PURPOSE: To describe and field-test a novel approach to prioritizing safety problems in rural primary care based on the method of Failure Modes and Effects Analysis. METHODS: A survey instrument designed to assess perceptions of medical error frequency, severity, and cause was administered anonymously to staff of 2 rural primary care practices in New York State. Responses were converted to quantitative hazard scores, which were used to make priority rankings of safety problems. Concordance analysis was conducted. RESULTS: Response rate was 94% at each site. Analysis yielded a list of priorities for each site. Comparison between staff groups (provider vs nursing vs administration), based on the top 10 priorities perceived by staff, showed 53% concordance at one site and 30% at the other. Concordance between sites was lower, at 20%. CONCLUSIONS: Initial field-testing of a Failure Modes and Effects Analysis approach in rural primary care suggests that it is feasible and can be used to estimate, based on staff perceptions, the greatest threats to patient safety in an individual practice so that limited resources can be focused appropriately. Higher concordance between staff within a practice than between practices lends preliminary support to the validity of the approach. PMID- 17397376 TI - Improving work environment perceptions for nurses employed in a rural setting. AB - CONTEXT: Effective recruitment and retention of professional nurses is a survival strategy for health care facilities, especially in rural areas. PURPOSE: This study examines the use of the Individual Workload Perception Scale to measure nurse satisfaction by a small rural hospital in order to make positive changes in the work environment for nurses. METHODS: Baseline work environment perceptions of nurses employed in a rural Kentucky hospital were assessed using the Individual Workload Perception Scale, a validated 38-item instrument. Nurses reviewed the results and brainstormed on potential interventions to address areas of concern. The 4 interventions selected for implementation by the nursing staff included (1) implementation of a shared decision making or governance model; (2) enhanced role of licensed practical nurses within the organization; (3) augmentation of administrative support on night and weekend shifts; and (4) utilization of wireless communication devices. After implementation of the interventions, staff nurse perceptions were reassessed using the same tool. FINDINGS: The follow-up survey revealed improvements in all areas measured by the Individual Workload Perception Scale, with the greatest improvement in the perception of the work environment noted among night nurses. The increase in positive work environment perception among these nurses, with greater than or equal to 11 years of professional experience, was statistically significant. CONCLUSIONS: Tools exist to support the development and evaluation of interventions to improve the work environment for nurses practicing in rural health care settings. By addressing issues of specific concern, both job satisfaction and retention of this talented pool of professionals can be enhanced. PMID- 17397377 TI - Evaluation of a medical school for rural doctors. AB - CONTEXT: Jichi Medical School (JMS) is the first and only medical school in Japan that was founded exclusively to graduate/prepare rural doctors. PURPOSE: To evaluate the long-term effect of JMS on the nationwide distribution of doctors. METHODS: Data from the Japanese population census of 1995 and from the Japanese physician census of 1994 were combined for use in this study. We extracted the JMS graduates from the physician census and compared the distribution of JMS graduates to that of non-JMS graduates. JMS graduates have an obligation to work in rural areas for 9 years after graduation. Therefore, we divided them into those doctors who were either "under rural duty" or "after rural duty." FINDINGS: JMS graduates were more likely than non-JMS physicians to practice in rural municipalities. The percentage of JMS graduates practicing under rural duty in communities meeting at least 1 of 4 possible criteria for being considered rural was 2.7 times greater than the percentage of non-JMS graduates in such communities. The percentage of JMS graduates practicing after rural duty in communities meeting at least 1 rural criterion was 2 times that of non-JMS graduates. The JMS graduates accounted for only 0.7% of all the physicians in Japan. However, they accounted for 4.2%, 1.5%, 1.8%, and 3.0% of the physicians in small population, remote, mountain, and medically underserved municipalities, respectively. CONCLUSIONS: The goal of JMS to produce rural doctors in Japan has made an impact on doctor distribution nationwide. PMID- 17397380 TI - Clinical and epidemiological features of canine leptospirosis in North Queensland. PMID- 17397381 TI - Lesions attributed to vaccination of sheep with Gudair for the control of ovine paratuberculosis: post farm economic impacts at slaughter. AB - OBJECTIVE: To determine i) the prevalence of lesions at the site of vaccination with Gudair, and ii) the incidence of discounting of slaughtered sheep carcasses due to the presence of these lesions. DESIGN: A survey of the prevalence of injection lesions and actual discounts applied to 20 consignments of vaccinates slaughtered in abattoirs in New South Wales. PROCEDURE: Consignments of sheep previously vaccinated against ovine Johne's disease (OJD) were assessed on the slaughter chain for the prevalence of vaccination site lesions and any costs associated with carcass trimming were estimated. In addition a telephone survey was conducted to determine the experiences and risk attitudes of 8 abattoirs in south east New South Wales likely to have previously processed vaccinates. RESULTS: The prevalence of lesions observed was 18% for adult (mutton) and 65% for lamb carcasses. The value of the trim removed was insignificant, the labour cost of its removal was nil and no carcass was downgraded to a lower value grade. CONCLUSION: Under the market conditions existing at the time the study was conducted, in sheep vaccinated at the recommended site high on the neck, it is unlikely that OJD vaccination site lesions will be a significant cost to producers or the processing industry and will represent only a very small proportion of the total cost of OJD control by vaccination. PMID- 17397383 TI - Skeletal and neurological malformations in pigs congenitally infected with Menangle virus. AB - OBJECTIVE: To describe the pathological findings in stillborn piglets and fetuses delivered by sows naturally infected with Menangle virus, a recently recognised Paramyxovirus. DESIGN: Observations of the gross and microscopic pathology of natural disease. PROCEDURE: Postmortem examinations were performed on 49 stillborn piglets, 35 mummified or semi-mummified full-term fetuses and 6 aborted fetuses from 20 litters at a 2600-sow intensive piggery in New South Wales during an outbreak of reproductive disease from June to September 1997. Body weights, crown-rump lengths and gross pathological changes were recorded. Tissues, including brain and spinal cord, were processed for histopathological examination. RESULTS: Litters with reduced numbers of live born piglets had mummified fetuses and stillborn piglets. Affected stillborn and aborted piglets frequently had arthrogryposis, craniofacial and spinal deformities, pulmonary hypoplasia and degeneration of the brain and spinal cord. Intranuclear and intracytoplasmic inclusion bodies were observed in neurones and other cells in the brain and spinal cord in association with extensive degeneration, necrosis, infiltration of macrophages and gliosis. CONCLUSIONS: In utero infection of piglets with Menangle virus is associated with severe skeletal and neurological malformations. PMID- 17397384 TI - Reproductive efficiency in grazing lactating dairy cows under a programmed reproductive management system. AB - OBJECTIVE: To evaluate the effectiveness of a reproductive management program consisting of combinations of Ovsynch/TAI and prostaglandin (PG) F(2alpha) treatments in Holstein dairy cows under a pasture-based dairying system. DESIGN: Field trial. PROCEDURE: A total of 1177 cows in 8 commercial dairy farms were randomly allocated to control and treatment groups. Treatment group cows received one of two interventions depending upon the number of days postpartum (DPP) before the planned start of breeding. Cows more than 50 DPP by the planned start of breeding received the Ovsynch/TAI treatment, consisting of gonadotrophin releasing hormone (GnRH) - PGF(2alpha)- GnRH plus timed artificial insemination. Cows between 40 and 50 DPP received a PGF(2alpha) treatment followed by oestrus detection and, if the cow was not seen in oestrus, the cow received a second PGF(2alpha) 14 days later. Control cows were submitted to twice a day heat detection followed by artificial insemination. The experimental period was the start of the breeding season plus 21 days for cows over 50 DPP at the start of breeding, and was 40-61 DPP for cows that calved later and passed their voluntary waiting period after the start of the breeding season. RESULTS: Submission rate was higher for the treated group than for the control group (84.9% vs. 55.1%; P < 0.0001), as was the conception rate (51.0% vs. 46.1%; P < 0.03). Due to farm variations, pregnancy rate was similar in both groups (38.5% vs. 28.2%; P > 0.1). Within the treated group, conception rate and pregnancy rate of the cows inseminated after a PGF(2alpha) were higher than for timed artificial inseminated cows (51.4% vs. 32.6%; P < 0.001), and (37.8% vs. 32.6%; P < 0.001). CONCLUSION: A programmed reproductive management protocol may improve reproductive efficiency in dairy farms with seasonal breeding, by increasing submission and conception rates at the beginning of the breeding season and/or at the end of the voluntary waiting period. Fertility of cows bred after a PGF(2alpha) synchronised heat was greater than after an Ovsynch/TAI protocol. PMID- 17397385 TI - Surgical repair of femoral fractures in New World camelids: five cases (1996 2003). AB - Five New World camelids were admitted to the Western College of Veterinary Medicine between 1996 and 2003 for evaluation of femoral fractures. There were three alpacas and two llamas. Four of the animals were female and three were less than 3 months of age. Fracture configurations consisted of distal physeal fractures (three), a comminuted diaphyseal/metaphyseal fracture, and a transverse diaphyseal fracture. Fractures were diagnosed with a combination of physical examination and radiographs in all cases. All five fractures were repaired with internal fixation and three animals were discharged from the hospital with fractures that healed. One cria underwent successful internal fixation but died from pulmonary oedema during recovery from anaesthesia. Postoperative complications were rare and limited to inadequate fracture stability in one alpaca and prolonged recovery to weight bearing in another. One llama with a comminuted metaphyseal fracture, repaired with a 4.5 mm dynamic compression plate, subsequently had catastrophic failure of the bone 17 days after surgery. Overall the clients were pleased with the outcome of discharged animals. Although femoral fractures are considered rare, they pose a unique opportunity for the large animal veterinarian to successfully achieve fracture union with the aid of internal fixation. PMID- 17397386 TI - Pleuropneumonia and pericarditis in a goat with isolation of Mycoplasma mycoides subspecies mycoides large colony. PMID- 17397387 TI - Dilation and torsion of the caecum in a cow caused by a trichobezoar. PMID- 17397389 TI - Effect of hypothermia on recovery from general anaesthesia in the dog. AB - OBJECTIVE: To discern the effects of anaesthesia protocols and decreasing core body temperature on time to recovery from general anaesthesia. MATERIALS AND METHODS: Healthy adult dogs undergoing desexing surgery were enrolled. More excitable dogs were premedicated with intramuscular acepromazine and morphine; calmer dogs were not premedicated. Anaesthesia was induced using halothane, isoflurane or sevoflurane delivered by mask, or by intravenous propofol, and maintained in standard fashion using one of the three inhalant agents. Thermostat controlled heat mats were used during surgical preparation and surgery. Oesophageal temperature was recorded throughout surgery. The time from cessation of anaesthetic administration until the dog successfully raised itself to sternal recumbency was considered the time of recovery. RESULTS: Sixty-nine dogs completed the study, 42 males anaesthetised for 60.4 +/- 20.5 min, and 27 females anaesthetised for 85.4 +/- 33.2 min. Oesophageal temperature at the end of surgery was 36.8 +/- 0.80 degrees C. Oesophageal temperature had a significant effect on recovery time, with lower temperatures contributing to slower recoveries. Premedication significantly lengthened recovery times. The choice of induction or maintenance anaesthetic agent had no effect on recovery time. DISCUSSION: Hypothermia is a common complication of general anaesthesia and surgery. Amongst other deleterious effects, it is associated with slower recovery from anaesthesia, likely due to a number of different mechanisms. PMID- 17397390 TI - Chondrosarcoma of the tongue of a horse. PMID- 17397392 TI - The HAL3-PPZ1 dependent regulation of nonsense suppression efficiency in yeast and its influence on manifestation of the yeast prion-like determinant [ISP(+)]. AB - The efficiency of stop codons read-through in yeast is controlled by multiple interactions of genetic and epigenetic factors. In this study, we demonstrate the participation of the Hal3-Ppz1 protein complex in regulation of read-through efficiency and manifestation of non-Mendelian anti-suppressor determinant [ISP(+)]. Over-expression of HAL3 in [ISP(+)] strain causes nonsense suppression, whereas its inactivation displays as anti-suppression of sup35 mutation in [isp( )] strain. [ISP(+)] strains carrying hal3Delta deletion cannot be cured from [ISP(+)] in the presence of GuHCl. Since Hal3p is a negative regulatory subunit of Ppz1 protein phosphatase, consequences of PPZ1 over-expression and deletion are opposite to those of HAL3. The observed effects are mediated by the catalytic function of Ppz1 and are probably related to the participation of Ppz1 in regulation of eEF1Balpha elongation factor activity. Importantly, [ISP(+)] status of yeast strains is determined by fluctuation in Hal3p level, since [ISP(+)] strains have less Hal3p than their [isp(-)] derivatives obtained by GuHCl treatment. A model considering epigenetic (possibly prion) regulation of Hal3p amount as a mechanism underlying [ISP(+)] status of yeast cell is suggested. PMID- 17397393 TI - Dynamic changes in the epigenomic state and nuclear organization of differentiating mouse embryonic stem cells. AB - Changes in nuclear organization and the epigenetic state of the genome are important driving forces for developmental gene expression. However, a strategy that allows simultaneous visualization of the dynamics of the epigenomic state and nuclear structure has been lacking to date. We established an experimental system to observe global DNA methylation in living mouse embryonic stem (ES) cells. The methylated DNA binding domain (MBD) and the nuclear localization signal (nls) sequence coding for human methyl CpG-binding domain protein 1 (MBD1) were fused to the enhanced green fluorescent protein (EGFP) reporter gene, and ES cell lines carrying the construct (EGFP-MBD-nls) were established. The EGFP-MBD nls protein was used to follow DNA methylation in situ under physiological conditions. We also monitored the formation and rearrangement of methylated heterochromatin using EGFP-MBD-nls. Pluripotent mouse ES cells showed unique nuclear organization in that methylated centromeric heterochromatin coalesced to form large clusters around the nucleoli. Upon differentiation, the organization of these heterochromatin clusters changed dramatically. Time-lapse microscopy successfully captured a moment of dramatic change in chromosome positioning during the transition between two differentiation stages. Thus, this experimental system should facilitate studies focusing on relationships between nuclear organization, epigenetic status and cell differentiation. PMID- 17397394 TI - Oxidative stress induces p53-dependent apoptosis in hepatoblastoma cell through its nuclear translocation. AB - Hepatoblastoma (HBL) is the most common malignant liver tumor in children. Since tumor suppressor p53 is rarely mutated in HBL, it remains unknown whether p53 could contribute to the hepatocarcinogenesis. In the present study, we have found for the first time that, like neuroblastoma (NBL), wild-type p53 was abnormally accumulated in the cytoplasm of the human HBL-derived Huh6 cells. In accordance with this notion, immunohistochemical analysis demonstrated that p53 is largely expressed in cytoplasm of human primary HBLs. In response to the oxidative stress, Huh6 cells underwent apoptotic cell death in association with the nuclear translocation of p53 and the transactivation of its target gene implicated in apoptotic cell death. siRNA-mediated knockdown of the endogenous p53 conferred the resistance of Huh6 cells to oxidative stress. Intriguingly, histone deacetylase inhibitor (nicotinamide) treatment strongly inhibited the oxidative stress-induced nuclear translocation of p53 as well as the p53-dependent apoptosis in Huh6 cells. In contrast to the previous observations, the cytoplasmic anchor protein for p53 termed Parc had undetectable effect on the cytoplasmic retention of p53. Collectively, our present results suggest that the abnormal cytoplasmic localization of p53 might contribute at least in part to the development of HBL. PMID- 17397395 TI - Molecular characterization of angiomotin/JEAP family proteins: interaction with MUPP1/Patj and their endogenous properties. AB - We have previously shown that MUPP1, which has an MRE domain and 13 PDZ domains, is expressed in epithelial cells and localize at tight junctions (TJs) and apical membranes. Using yeast two-hybrid screening, we found here that MUPP1 interacts with angiomotin (Amot), JEAP/Amot-like 1 and MASCOT/Amot-like 2, which we refer to as Amot/JEAP family proteins. PDZ2 and -3 were responsible for MUPP1's interaction with Amot and MASCOT, whereas only PDZ3 was responsible for its interaction with JEAP. All the Amot/JEAP family proteins also interacted with Patj, a close relative of MUPP1. The C-terminal PDZ-binding motives of the Amot/JEAP family were required for these interactions. We successfully generated specific antibodies for these proteins and analyzed the endogenous molecular properties of the family in parallel. Immunofluorescence microscopy of cultured epithelial cells showed that in subcellular distribution, the Amot/JEAP family proteins were indistinguishable; they were apparent at TJs as well as apical membranes, and mostly co-localized with MUPP1. They were also located at TJs in several mouse tissues, but each protein showed a distinct tissue distribution. In biochemical fractionation assays, the Amot/JEAP family behaved not as transmembrane but as peripheral membrane proteins. Unexpectedly, the PDZ-binding motives were not necessarily required for their localization to TJs, and dominant negative MUPP1 or Patj did not affect the localization of Amot/JEAP family proteins, suggesting that the interaction with MUPP1/Patj is not necessarily responsible for their proper subcellular distribution. PMID- 17397396 TI - Heat shock transcription factor 1 down-regulates spermatocyte-specific 70 kDa heat shock protein expression prior to the induction of apoptosis in mouse testes. AB - Expression of constitutively active heat shock transcription factor 1 (HSF1) in mouse spermatocytes induces apoptosis and leads to male infertility. We report here that prior to the onset of massive apoptosis caused by expression of active HSF1 in spermatocytes a marked reduction in spermatocyte-specific Hsp70.2 mRNA and protein levels occurs. In addition, HSP70.2 protein relocalizes from a predominant cytoplasmic to a nuclear position in developing spermatocytes that express active HSF1. Later in the developmental stages, cells undergoing HSF1 induced apoptosis essentially lack the HSP70.2 protein. The down-regulation of Hsp70.2 gene expression by HSF1 is paradoxical because HSF1 is the prototypical activator of HSP genes. Furthermore, HSF1-mediated repression neither involved a heat shock element (HSE)-like sequence adjacent to the Hsp70.2 gene nor were Hsp70.2 promoter sequences associated directly with HSF1. Interestingly, other spermatocyte- and spermatid-specific transcripts are also down-regulated in testes of transgenic mice expressing active HSF1, suggesting involvement of a putative HSF1-dependent block of development of spermatogenic cells. Importantly however, transcription of the Hsp70.2 gene is down-regulated in testes of wild type mice subjected to a hyperthermia that induces transient activation of HSF1, indicating that the spermatocyte-specific activity of HSF1 might misdirect a network of transcription factors required for proper regulation of Hsp70.2. PMID- 17397397 TI - The N-terminal regions of eukaryotic acidic phosphoproteins P1 and P2 are crucial for heterodimerization and assembly into the ribosomal GTPase-associated center. AB - Acidic phosphoproteins P1 and P2 form a heterodimer and play a crucial role in assembly of the GTPase-associated center in eukaryotic ribosomes and in ribosomal interaction with translation factors. We investigated the structural elements within P1 and P2 essential for their dimerization and for ribosomal function. Truncation of the N-terminal 10 amino acids in either P1 or P2 and swapping of the N-terminal 10 amino acid sequences between these two proteins disrupted their dimerization, binding to P0 and P0 binding to rRNA. In contrast, truncation of the C-terminal halves of P1 and P2 as well as swapping of these parts between them gave no significant effects. The protein dimers containing the C-terminal truncation mutants or swapped variants were assembled with P0 onto Escherichia coli 50 S subunits deficient in the homologous protein L10 and L7/L12 and gave reduced ribosomal activity in terms of eukaryotic elongation factor dependent GTPase activity and polyphenylalanine synthesis. The results indicate that the N terminal 10 amino acid sequences of both P1 and P2 are crucial for P1-P2 heterodimerization and for their functional assembly with P0 into the GTPase associated center, whereas the C-terminal halves of P1 and P2 are not essential for the assembly. PMID- 17397398 TI - Synaptotagmin VII modulates the kinetics of dense-core vesicle exocytosis in PC12 cells. AB - In our previous study, we showed that PC12 cell lines stably expressing synaptotagmin (Syt) VII have greater ability to release hormones Ca(2+) dependently than the original PC12 cells. However, the precise molecular mechanism of the enhancement of hormone secretion by Syt VII has never been elucidated. In this study, we established a PC12 cell line that stably expresses Syt VII-green fluorescent protein (Syt VII-GFP) or its Ca(2+)-binding-site deficient mutant (D172N/D303N substitutions; Syt VII-DN-GFP), and examined the effect of Syt VII-GFP expression on the kinetics of dense-core vesicle exocytosis by total internal reflection fluorescence (TIRF) microscopy. Both Syt VII-GFP and Syt VII-DN-GFP co-localized well with dense-core vesicle markers, monomeric red fluorescent protein (mRFP)-tagged neuropeptide Y (NPY-mRFP) and cyan fluorescent protein (CFP)-tagged tissue plasminogen activator (tPA-CFP). Expression of Syt VII-GFP enhanced the number of dense-core vesicle exocytotic events, whereas expression of Syt VII-DN-GFP or knockdown of Syt VII-GFP with specific small interfering RNA (siRNA) attenuated the number of exocytotic events. Monitoring individual tPA-CFP release events revealed that "full release" events are increased in Syt VII-GFP-expressing cells, but not in Syt VII-DN-GFP-expressing or Syt VII-silenced cells. Our data indicate that Syt VII modulates the kinetics of Ca(2+)-dependent dense-core vesicle exocytosis in neuroendocrine PC12 cells, possibly by modulating fusion pore opening. PMID- 17397399 TI - Glycosylation and palmitoylation of Wnt-3a are coupled to produce an active form of Wnt-3a. AB - Wnt-3a is a representative ligand that activates the beta-catenin-dependent pathway in Wnt signaling and is modified with glycans and palmitate. In this study, we analyzed the relationship between glycosylation and lipidation of Wnt 3a. Secretion of a Wnt-3a mutant that lacks glycosylation (Wnt-3a NQ) was impaired. Wnt-3a C77A, which lacks palmitoylation at Cys77, was secreted with similar efficiency to wild-type Wnt-3a (Wnt-3a WT), but did not induce the internalization of low-density lipoprotein receptor-related protein 6 (LRP6). Furthermore, removal of palmitate from Wnt-3a suppressed the ability to bind to its receptors Frizzled8 and LRP6. Wnt-3a C77A was glycosylated to an extent similar to Wnt-3a WT, while Wnt-3a NQ was not modified with palmitate. Expression of porcupine, which is a putative acyltransferase, enhanced palmitoylation of Wnt 3a WT greatly, but that of Wnt-3a NQ slightly. While Wnt-3a WT was present in both the endoplasmic reticulum (ER) and Golgi, Wnt-3a NQ was located to the ER only. Furthermore, Wnt-3a was not palmitoylated but was glycosylated in the cells treated with Brefeldin A, which inhibits transport of vesicles from the ER to the Golgi. These results indicate that glycosylation of Wnt-3a precedes palmitoylation and that both modifications are necessary for secretion of an active Wnt-3a. PMID- 17397400 TI - The PI3K-Akt pathway promotes microtubule stabilization in migrating fibroblasts. AB - Directed cell migration is controlled by extracellular cues such as growth factors/chemokines and extracellular matrix. In a migrating cell, a subset of microtubules becomes stabilized, and this stabilization is implicated in the establishment and maintenance of cell polarity. It is still not fully understood, however, how extracellular cues regulate the dynamics of microtubules. Here we show that the PI3K-Akt signaling pathway plays a pivotal role in growth factor regulation of microtubule stability. Treatment of NIH 3T3 fibroblasts with platelet-derived growth factor (PDGF) increases the amount of stabilized microtubules, and this increase is abrogated by the addition of a PI3K inhibitor or by expression of a dominant-negative form of Akt (DN-Akt), but not by the addition of a MEK inhibitor. Expression of an active form of Akt slightly increases the bulk amount of stabilized microtubules. Stabilization of microtubules induced in edge cells in the wounded monolayer culture is also attenuated by the PI3K inhibitor treatment or by expression of DN-Akt. Given that Akt is activated at the leading edge of a migrating cell and plays an essential role in directed cell migration, these results reveal a novel mechanism linking extracellular cues to directed cell migration, namely Akt regulation of microtubule stability. PMID- 17397401 TI - ABIM recertification in gastroenterology: ACG's contribution. PMID- 17397402 TI - Propofol use under the direction of trained gastroenterologists: an analysis of the medicolegal implications. PMID- 17397403 TI - Circumcision kit. PMID- 17397404 TI - Limited diagnostic value of laryngopharyngeal lesions in patients with gastroesophageal reflux during routine upper gastrointestinal endoscopy. AB - BACKGROUND AND AIMS: There is growing evidence that gastroesophageal reflux disease (GERD) may cause typical laryngeal/pharyngeal lesions secondary to tissue irritation. The prevalence of those lesions in GERD patients is not well established. The aim of this study was to evaluate the prevalence of GERD signs in the laryngopharyngeal area during routine upper gastrointestinal endoscopy. METHODS: Between July 2000 and July 2001, 1,209 patients underwent 1,311 upper gastrointestinal endoscopies and were enrolled in this study. The structured examination of the laryngopharyngeal area during upper gastrointestinal endoscopy was videotaped for review by three gastroenterologists and one otorhinolaryngologist, blinded to the endoscopic esophageal findings. From the 1,209 patients enrolled in this prospective study, all patients (group I, N = 132) with typical endoscopical esophageal findings of GERD (Savary-Miller I-IV) were selected. The sex- and age-matched control group II (N = 132) underwent upper gastrointestinal endoscopy for different reasons, had no reflux symptoms, and had normal esophagoscopy. RESULTS: In the two groups of patients, we found no difference in the prevalence of abnormal interarytenoid bar findings (32%vs 32%), arytenoid medial wall erythema (47%vs 43%), posterior commissure changes (36%vs 34%), or posterior cricoid wall edema (1%vs 3%). The only difference was noted in the posterior pharyngeal wall cobblestoning (66%vs 50%, P= 0.004). CONCLUSION: The results of this large systematic investigation challenge the diagnostic specificity of laryngopharyngeal findings attributed to gastroesophageal reflux. PMID- 17397405 TI - Are there specific laryngeal signs for gastroesophageal reflux disease? PMID- 17397406 TI - Capsule endoscopy versus computed tomographic or standard angiography for the diagnosis of obscure gastrointestinal bleeding. AB - BACKGROUND AND AIMS: Capsule endoscopy (CE) is superior to push enteroscopy and small bowel barium radiography in detecting the source of obscure GI bleeding. We now compared whether CE has a superior diagnostic yield than CT angiography (CTA) or standard mesenteric angiography (ANGIO) in patients with obscure GI bleeding. METHODS: From June 2004 to October 2005, consecutive patients admitted for OGIB underwent both CTA and ANGIO, followed by CE, performed blindly by independent examiners within the next 7 days. The primary end point of the study was the diagnostic yield for each technique, defined as the frequency of detection of lesions with a high probability of bleeding. RESULTS: Twenty-eight patients (16 men and 12 women, mean age 74 +/- 2 yr) with OGIB (overt bleeding in 20 cases and chronic occult in 8) were prospectively evaluated. CTA or standard angiography could be performed in 25 of 28 patients (applicability 86%), because of contrast allergy (1 patient) and chronic renal failure (2 patients). A source of bleeding was detected by CE in a greater proportion of patients, 72% (18 of 25, 95% CI 50.6-87.9%), than CTA, 24% (6 of 25, 95% CI 9.4-45.1%, P= 0.005 vs CE), or ANGIO, 56% (14 of 25, 95% CI 34.9-75.6%, P= NS). Similarly, CE was able to diagnose 100% of patients diagnosed by CTA and 86% of patients diagnosed by ANGIO. Moreover, CE was positive in 12 of 19 (63%) negative cases on CTA and in 6 of 11 (55%) negative cases on ANGIO. As a result of the CE findings, therapeutic intervention was undertaken in 9 of 19 (47%) patients with positive results. CONCLUSION: CE detects more lesions than CTA or standard mesenteric angiography in patients with obscure GI bleeding and has a therapeutic impact in almost half of the patients with positive findings. PMID- 17397407 TI - Hepatitis B virus infection in 6,130 unvaccinated Korean-Americans surveyed between 1988 and 1990. AB - OBJECTIVES: During the past decades, the influx of immigrants from hepatitis B virus (HBV) endemic regions has brought significant changes in the prevalence of HBV-associated liver diseases and hepatocellular carcinoma (HCC) in the United States. Our program, which was intended to identify those in need of hepatitis B vaccination, helped us to learn of the natural history of HBV infection in Korean Americans. METHODS: Between November of 1988 and May 1990, we screened 6,130 Korean Americans in the eastern United States for HBV infection. RESULTS: The overall hepatitis B surface antigen (HBsAg) (+) rate was 6.1%, with 8.0% for males and 4.4% for females. The carrier rate peaked in subjects between the ages of 21 and 40 yr. The HBsAg (+) rate for 452 U.S.-born children was lower (2.7%) than that of 623 Korean-born (5.5%). None received hepatitis B immune-globulin or HBV vaccination. The vertical transmission rate was 30.3% in children born to HBsAg (+) mothers and 100% in those born to hepatitis B e antigen (HBeAg) positive mothers. In contrast, the paternal transmission rate was low; 10.3% in children with HBsAg (+) fathers and 19.2% in those with HBeAg (+) fathers. Another significant observation was the unexpected finding of ongoing liver diseases in incidentally identified carriers. Evaluation of 139 asymptomatic adult carriers revealed that 42% had elevated liver enzymes and 11% had already developed liver cirrhosis. CONCLUSION: These findings strongly suggest the need for active HBV screening of immigrants from endemic regions and, most importantly, the need for careful monitoring of the carriers. PMID- 17397408 TI - The degree of breath methane production in IBS correlates with the severity of constipation. AB - BACKGROUND: Recent work has demonstrated that among irritable bowel syndrome (IBS) subjects, methane on lactulose breath test (LBT) is nearly universally associated with constipation predominance. This work has been based on subjective constipation outcomes. In this study, methane is compared to constipation in another population of IBS subjects with constipation being determined both subjectively and objectively. METHODS: A nested study was conducted in subjects enrolled in a double-blind randomized placebo-controlled study. After consent, subjects were asked to complete a stool diary for 7 days. This included logging of all bowel movements that week as well as documenting the stool consistency for each during the same period using the Bristol Stool Score. After 7 days, subjects were asked to rate their symptoms on a visual analogue scale (VAS) score (0-100 mm) for diarrhea and constipation. They then had an LBT to evaluate both methane and hydrogen profiles over 180 min. Subjects with methane were compared to those without methane for Bristol Stool Score, stool frequency, as well as VAS scores for diarrhea and constipation. The degree of constipation was then compared to the quantity of methane production on LBT based on area under the curve. RESULTS: Among 87 subjects, 20 (23.8%) produced methane. IBS subjects with methane had a mean constipation severity of 66.1 +/- 36.7 compared to 36.2 +/- 30.8 for nonmethane producers (P < 0.001). The opposite was noted for diarrhea (P < 0.01). On LBT, the quantity of methane seen on breath test was directly proportional to the degree of constipation reported (r = 0.60, P < 0.01). In addition, greater methane production correlated with a lower stool frequency (r =-0.70, P < 0.001) and Bristol Stool Score (r =-0.58, P < 0.01). CONCLUSION: Methane on LBT is associated with constipation both subjectively and objectively. The degree of methane production on breath test appears related to the degree of constipation. PMID- 17397409 TI - Gas distribution within the human gut: effect of meals. AB - BACKGROUND AND AIMS: Patients frequently complain of gas symptoms precipitated by meals, but the effect of early digestion on intestinal gas content remains unknown. Our aim was to determine the influence of meals on intestinal gas volume and distribution. METHODS: First, we developed a CT image analysis program, based on independent software modules, to measure gas content within the gut. The system was validated in nine healthy subjects by taking helical abdominal CT scans before and after rectal infusion of known volumes of air (100-400 mL). In 15 healthy subjects, intestinal gas distribution was measured in fast and early postcibal CT scans. The postcibal scan was taken 99 +/- 22 minutes after a 597 +/ 57 kcal meal. RESULTS: The volume of gas infused per rectum was detected with an accuracy of 100.4 +/- 3.0%. During fasting, intestinal gas volume was 94 +/- 7 mL (excluding two extreme outliers). After the meal, gas content within the gut increased by 64.7% (up to 149 +/- 21 mL, P < 0.01 vs fast) and the increment occurred in the colon (59 +/- 9 mL precibal vs 121 +/- 20 mL postcibal, P < 0.001), while other gut compartments remained unchanged. CONCLUSION: Ingestion of a meal activated gas metabolism and increased gas content within the gut. The increment occurred early, presumably prior to colonic fermentation of food substrates and was localized in the distal gut, suggesting that gas had a proximal origin and was propelled caudally. PMID- 17397410 TI - High-resolution manometry in the evaluation of anorectal disorders: a simultaneous comparison with water-perfused manometry. AB - INTRODUCTION: High-resolution manometry (HRM) combined with novel interpretive software allows for the interpolation of manometric recordings into highly detailed topographical plots of intraluminal pressure events relative to time and location. To date, HRM has been used primarily in the study of esophageal disorders and has been shown to provide greater physiologic resolving power compared with standard manometric techniques. This preliminary feasibility study is the first report evaluating HRM and simultaneously performed water-perfused manometry (WPM) in patients referred for anorectal manometry (ARM). METHODS: Consecutive patients referred for ARM underwent simultaneous WPM and HRM. WPM was performed using a 6-cm sleeve assembly with side-holes spaced at 3-cm intervals. HRM was performed using the ManoScan system (Sierra Scientific Instruments, Los Angeles, CA) consisting of a 36-channel catheter with sensors spaced at 1-cm intervals. Space-time pressure data were displayed in topographic form using ManoView analysis software that is part of the ManoScan system. Measurements of anal sphincter pressure at rest, during voluntary contraction, and during 40-mL rectal balloon distension were compared. RESULTS: A total of 29 patients were studied. Indications for manometry were constipation (19), incontinence (7), and fecal soilage (3). Simultaneously recorded resting, squeeze, and relaxation pressures showed the two methods to be significantly correlated although anal sphincter pressures recorded by HRM tended to be higher than those recorded with WPM. HRM provided greater resolution of the intraluminal pressure environment of the anorectum. CONCLUSIONS: These preliminary observations demonstrate that anorectal HRM highly correlated with WPM measurements and provided greater anatomic detail. PMID- 17397411 TI - "Demand the data! Your patients will ask and your practice will need it". AB - The efficacy of endoscopic screening for colorectal neoplasia is strongly influenced by adequacy of the preparation and the quality of the exam. Past studies have emphasized the association of diminished polyp detection with overly fast withdrawal from the cecum. In this issue of the journal, Chen and Rex present data more directly associating differences in the rate of polyp detection with the individual endoscopist, demonstrating that some do a better job than others. Why this is so is unclear. Further data are needed regarding the causes and means to reduction of inter-endoscopist differences. Few units report and few endoscopists actually know their personal rates for polyp detection. As with cecal intubation rates and withdrawal times, provision of personal and group data would undoubtedly enhance performance. PMID- 17397412 TI - Strategies for managing anemia in hepatitis C patients undergoing antiviral therapy. AB - Anemia is a common side effect that begins soon after the initiation of peginterferon/ribavirin in the treatment of hepatitis C virus (HCV) infection. It can cause symptoms that negatively impact quality of life (QOL) and is the most common reason for reducing the dose and temporarily or permanently discontinuing ribavirin. Such dose modifications have been shown to reduce the efficacy of treatment. Administering erythropoietin can improve anemia caused by peginterferon and ribavirin therapy and is more effective than dose reduction at improving QOL during treatment. However, erythropoietin, which is not approved by the U.S. Food and Drug Administration (FDA) for use in patients with HCV infection, adds another parenteral drug to the patient's treatment regimen, and is associated with additional costs, inconvenience, and potential side effects. A new ribavirin analog, viramidine, is expected to be associated with a lower incidence of anemia and, if proven effective, may eventually be substituted for ribavirin in combination with peginterferon to treat chronic hepatitis C. In the meantime, physicians must make the best possible use of the available options for managing anemia, especially in select patient groups who are most at risk for anemia and its complications. PMID- 17397414 TI - Upper endoscope for colonoscopy: evidence is still too sparse to utilize. PMID- 17397415 TI - How deep should "deep sedation" be? PMID- 17397418 TI - Good colon preparation guarantees a higher diagnostic yield of colonoscopy-timing is most crucial. PMID- 17397419 TI - Type 2 diabetes mellitus, smoking, and colorectal cancer. PMID- 17397420 TI - Amyloidoma of the esophagus. PMID- 17397421 TI - Argument on statement 46 in the Montreal definition of gastroesophageal reflux disease: does reflux laryngitis patient always have heartburn? PMID- 17397425 TI - Kenosha County falls prevention study: a randomized, controlled trial of an intermediate-intensity, community-based multifactorial falls intervention. AB - OBJECTIVES: To decrease the rate of falls in high-risk community-dwelling older adults. DESIGN: Randomized, controlled trial. SETTING: Community-based. PARTICIPANTS: Three hundred forty-nine adults aged 65 and older with two falls in the previous year or one fall in the previous 2 years with injury or balance problems. INTERVENTION: Subjects received two in-home visits from a trained nurse or physical therapist who assessed falls risk factors using an algorithm. The intervention consisted of recommendations to the subject and their primary physician, referrals to physical therapy and other providers, 11 monthly telephone calls, and a balance exercise plan. Control subjects received a home safety assessment. MEASUREMENTS: The primary outcome was rate of falls per year in the community. Secondary outcomes included all-cause hospitalizations and nursing home admissions per year. RESULTS: There was no difference in rate of falls between the intervention and control groups (rate ratio (RR)=0.81, P=.27). Nursing home days were fewer in the intervention group (10.3 vs 20.5 days, P=.04). Intervention subjects with a Mini-Mental State Examination (MMSE) score of 27 or less had a lower rate of falls (RR=0.55; P=.05) and, if they lived with someone, had fewer hospitalizations (RR=0.44, P=.05), nursing home admissions (RR=0.15, P=.003), and nursing home days (7.5 vs 58.2, P=.008). CONCLUSION: This multifactorial intervention did not decrease falls in at-risk community-living adults but did decrease nursing home utilization. There was evidence of efficacy in the subgroup who had an MMSE score of 27 or less and lived with a caregiver, but validation is required. PMID- 17397426 TI - A randomized, controlled trial of fall prevention programs and quality of life in older fallers. AB - OBJECTIVES: To compare the effects of three fall-prevention programs (education (ED), home safety assessment and modification (HSAM), and exercise training (ET)) on quality of life (QOL), functional balance and gait, activities of daily living (ADLs), fear of falling, and depression in adults aged 65 and older. DESIGN: A 4 month randomized trial. SETTING: Randomized, controlled trial. PARTICIPANTS: One hundred fifty participants who had experienced a recent fall. MEASUREMENTS: QOL was assessed according to the brief version of the World Health Organization Quality of Life instrument (WHOQOL-BREF), functional balance and gait according to functional reach and Tinetti balance and gait, ADLs according to the Older Americans Resources and Services questionnaire, fear of falling according to a visual analog scale, and depression level according to the Geriatric Depression Scale. RESULTS: The score changes for the ET group were 2.1 points greater on the physical domain (95% confidence interval (CI)=-1.2-5.3), 3.8 points greater on the psychological domain (95% CI=0.7-7.0), and for the WHOQOL-BREF, 3.4 points greater on the social domain (95% CI=0.7-6.1) and 3.2 points greater on the environmental domain (95% CI=0.6-5.7) than for the ED group. The score change for each domain of the WHOQOL-BREF for the HSAM group was greater than that for the ED group, although these results were not statistically significant. The ET group also had greater improvements in functional reach, Tinetti balance and gait, and fear of falling than the ED group. CONCLUSION: The QOL outcome supports the superiority of ET over the other two interventions in older people who have recently fallen. This finding also parallels those gathered from the functional measures. PMID- 17397427 TI - Maximum potential preventive effect of hip protectors. AB - OBJECTIVES: To estimate the maximum potential preventive effect of hip protectors in older persons living in the community or homes for the elderly. DESIGN: Observational cohort study. SETTING: Emergency departments in the Netherlands. PARTICIPANTS: Hip fracture patients aged 70 and older who visited the emergency departments of five hospitals in the Netherlands (n=520). MEASUREMENTS: Using the risk score of the Dutch Guidelines for Osteoporosis, how many patients had a high risk for fractures was retrospectively assessed. In addition, the circumstances of the hip fracture were assessed (n=299). Four factors were specified that might influence the maximum potential preventive effect of hip protectors: (1) hip fracture occurred in persons having a low risk, (2) hip fracture was not the consequence of a fall, (3) hip fracture occurred during circumstances that preclude the use of hip protectors, and (4) hip fracture occurred during the night. RESULTS: When providing hip protectors to women at high risk of fractures, 48.2% of all hip fractures could have been prevented. CONCLUSION: Many hip fractures occur in persons with a low risk for hip fracture or under circumstances that preclude the use of hip protectors. It was estimated that the maximum potential preventive effect of hip protectors is approximately 50% in older women living in the community or homes for the elderly. The actual preventive effect will be lower and depends on the acceptance and effectiveness of hip protectors and adherence to wearing them. PMID- 17397428 TI - Augmenting immune responses to varicella zoster virus in older adults: a randomized, controlled trial of Tai Chi. AB - OBJECTIVES: To evaluate the effects of a behavioral intervention, Tai Chi, on resting and vaccine-stimulated levels of cell-mediated immunity (CMI) to varicella zoster virus (VZV) and on health functioning in older adults. DESIGN: A prospective, randomized, controlled trial with allocation to two arms (Tai Chi and health education) for 25 weeks. After 16 weeks of intervention, subjects were vaccinated with VARIVAX, the live attenuated Oka/Merck VZV vaccine licensed to prevent varicella. SETTING: Two urban U.S. communities between 2001 and 2005. PARTICIPANTS: A total of 112 healthy older adults aged 59 to 86. MEASUREMENTS: The primary endpoint was a quantitative measure of VZV-CMI. Secondary outcomes were scores on the Medical Outcomes Study 36-item Short-Form Health Survey (SF 36). RESULTS: The Tai Chi group showed higher levels of VZV-CMI than the health education group (P<.05), with a significant rate of increase (P<.001) that was nearly twice that found in the health education group. Tai Chi alone induced an increase in VZV-CMI that was comparable in magnitude with that induced by varicella vaccine, and the two were additive; Tai Chi, together with vaccine, produced a substantially higher level of VZV-CMI than vaccine alone. The Tai Chi group also showed significant improvements in SF-36 scores for physical functioning, bodily pain, vitality, and mental health (P<.05). CONCLUSION: Tai Chi augments resting levels of VZV-specific CMI and boosts VZV-CMI of the varicella vaccine. PMID- 17397429 TI - Influence of comorbid conditions on long-term mortality after pneumonia in older people. AB - OBJECTIVES: To test the hypothesis that increased long-term mortality after hospitalization for community-acquired pneumonia (CAP) is independent of comorbid conditions. DESIGN: Prospective observational cohort study in metropolitan areas. SETTING: Memphis, Tennessee, and Pittsburgh, Pennsylvania. PARTICIPANTS: Three thousand seventy-five subjects aged 70 to 79 over 5.2 years. MEASUREMENTS: Unadjusted and adjusted mortality from an initial hospitalization for CAP were compared with mortality from different causes of hospitalization, including cancer, fracture, congestive heart failure (CHF), cerebrovascular accident (CVA), and other causes. Demographics, smoking, nutritional markers, functional status, inflammatory markers, and chronic health conditions were adjusted for. RESULTS: Of the 106 subjects hospitalized for CAP, 22 (20.8%) and 38 (35.8%) died at 1 and 5 years. Subjects hospitalized with CAP had higher mortality than nonhospitalized subjects (adjusted odds ratio (OR)=7.8, 95% confidence interval (CI)=4.2-14.4). One- and 5-year mortality after CAP hospitalization were higher than mortality from other causes requiring hospitalization and remained unchanged in multivariable analysis (adjusted OR=3.5, 95% CI=1.5-8.1; adjusted OR=5.6, 95% CI=2.8-11.2, respectively). One- and 5-year mortality after hospitalization for CAP were similar to or higher than mortality after an initial hospitalization for CHF, CVA, or fracture. Rehospitalization was common in subjects hospitalized for CAP and may explain greater long-term mortality. CONCLUSION: In this high functioning cohort of older persons, an initial hospitalization for CAP was associated with greater long-term mortality, independent of prehospitalization comorbid conditions. Hospitalization for CAP has as serious a prognosis as hospitalization for CHF, stroke, or major fracture. PMID- 17397430 TI - Protective association between neighborhood walkability and depression in older men. AB - OBJECTIVES: To evaluate the association between neighborhood walkability and depression in older adults. DESIGN: Cross-sectional analysis using data from Adult Changes in Thought (ACT), a prospective, longitudinal cohort study. SETTING: King County, Washington. PARTICIPANTS: Seven hundred forty randomly selected men and women aged 65 and older, cognitively intact, living in the same home for at least 2 years. MEASUREMENTS: Depressive symptoms were measured with the Center for Epidemiologic Studies Depression Scale. The Walkable and Bikable Communities Project provided objective data predicting the probability of walking at least 150 minutes per week in a particular neighborhood. ACT data were linked at the individual level via a geographic information system to this walkability score using buffer radii of 100, 500, and 1,000 meters around the subject's home. Multiple regression analysis tests were conducted for associations between the buffer-specific neighborhood walkability score and depressive symptoms. RESULTS: There was a significant association between neighborhood walkability and depressive symptoms in men when adjusted for individual-level factors of income, physical activity, education, smoking status, living alone, age, ethnicity, and chronic disease. The odds ratio for the interquartile range (25th to 75th percentile) of walkability score was 0.31 to 0.33 for the buffer radii (P=.02), indicating a protective association with neighborhood walkability. This association was not significant in women. CONCLUSION: This study demonstrates a significant association between neighborhood walkability and depressive symptoms in older men. Further research on the effects of neighborhood walkability may inform community-level mental health treatment and focus depression screening in less-walkable areas. PMID- 17397431 TI - Predisposing factors and precipitants for bathing disability in older persons. AB - OBJECTIVES: To identify potential predisposing factors and precipitants for bathing disability. DESIGN: Prospective cohort study. SETTING: Greater New Haven, Connecticut. PARTICIPANTS: Seven hundred fifty-four community-living residents aged 70 and older. MEASUREMENTS: Potential predisposing factors were measured every 18 months for 6 years during comprehensive home-based assessments. Participants were followed with monthly telephone interviews for a median of 61 months to ascertain exposure to potential precipitants, which included illnesses and injuries leading to hospitalization or restricted activity, and to determine the occurrence of persistent disability in bathing. RESULTS: Over the course of nearly 8 years, 333 (44.2%) participants developed persistent disability in bathing. After accounting for age, sex, and race, nine predisposing factors were independently associated with persistent bathing disability. The strongest associations were found for inability to rise from a chair, low bathing self efficacy, and low physical activity, each of which more than doubled the risk of persistent bathing disability. After the potential precipitants were added to the final model, the associations between the predisposing factors and persistent bathing disability were only modestly diminished. The effects of the precipitants on persistent bathing disability were large, with multivariable hazard ratios of 24.6 for hospitalization and 3.01 for restricted activity only. The relative contributions of the predisposing factors and precipitants to the development of persistent bathing disability were 47.7% and 27.1%, respectively. CONCLUSION: Disability in bathing is attributable to a combination of predisposing factors that make community-living older persons vulnerable and intervening illnesses or injuries that act as precipitants. These factors and precipitants may be suitable targets for the prevention of bathing disability. PMID- 17397432 TI - Development, worsening, and improvement of diabetic microangiopathy in older people: six-year prospective study of patients under intensive diabetes control. AB - OBJECTIVES: To examine retinopathy and nephropathy in elderly patients with diabetes mellitus (DM) under intensive multifactorial DM control. DESIGN: Six year interventional observation study. SETTING: Multicenter study including four hospitals. PARTICIPANTS: Four hundred thirteen elderly (> or = 65) patients with type 2 DM attending each hospital for 1 year or longer; those receiving hemodialysis or with uncured malignancy were excluded. MEASUREMENTS: Development, worsening, and improvement of retinopathy and nephropathy and respective risk factors. RESULTS: The mean baseline hemoglobin (HbA1c), blood pressure (BP), and total cholesterol were 6.8%, 137/74 mmHg, and 5.13 mmol/L, respectively. Retinopathy developed in 45 of 168 (27%) patients and, of 63 with nonproliferative retinopathy, worsened and improved in 11 (17%) and 23 (37%), respectively. Nephropathy developed in 53 of 227 (23%) patients and improved in 13 of 51 (25%) having it baseline. The mean change in glomerular filtration rate (DeltaGFR, baseline GFR-GFR at the end of the study period) in those with nephropathy at baseline was 21.5 mL/min. HbA1c was related to development of retinopathy (P=.001, odds ratio (OR)=1.91), and serum creatinine (P=.03, OR=1.02), systolic BP (SBP) (P=.03, OR=1.22), and prior stroke (P=.005, OR=3.21) were related to development of nephropathy. In patients with nephropathy at baseline, SBP (P=.03, Spearman's rho (rho)=0.310), total cholesterol (P=.01, rho=0.361), and low-density lipoprotein cholesterol (P=.03, rho=0.322) were correlated with DeltaGFR. CONCLUSION: In elderly patients under intensive control for DM, the outcome of microangiopathy is favorable. Modifiable risk factors were hyperglycemia for development of retinopathy and hypertension and hypercholesterolemia for development or worsening of nephropathy; prior stroke was an unmodifiable risk factor for development of nephropathy. PMID- 17397433 TI - Testosterone, sex hormone-binding globulin, and frailty in older men. AB - OBJECTIVES: To determine whether testosterone (T) levels are associated with frailty or its components. DESIGN: Population-based cohort study conducted in three waves (T1: 1987-1989, T2: 1995-1997, T3: 2002-2004). SETTING: Communities in the Boston, Massachusetts, area. PARTICIPANTS: Six hundred forty-six men aged 50 to 86 at T(3) with complete data on frailty components and hormone measurements. MEASUREMENTS: The frailty phenotype was defined as the presence of three or more of the following: weight loss, exhaustion, low physical activity, slowness, and weakness. Men were classified as frail (> or = 3 components), intermediate (1-2 components), and nonfrail (0 components). Whether total and free T or sex hormone-binding globulin (SHBG) levels were associated cross sectionally with frailty and with degree of frailty was determined. Potential confounders such as age, chronic disease, lifestyle factors, diet, and physical activity were considered. RESULTS: No association was observed between total or free T and the frailty phenotype after adjusting for confounders. Conversely, a significant association was observed between SHBG and frailty phenotype with an adjusted odds ratio of 1.25 (95% confidence interval=1.06-1.46) per 10-nM increase in SHBG levels. Associations between hormones and degree of frailty were similar to those for overall frailty. Of frailty components, grip strength and physical activity, but not exhaustion, slow walking, or weight loss, were associated with total T levels, whereas SHBG was related to weight loss, exhaustion, and physical activity. CONCLUSION: Total and free T levels were not associated with frailty phenotype, but SHBG was. Furthermore, T and SHBG levels were associated with some, but not all, components of frailty. Therefore, T trials in older men should focus on men experiencing decreases in strength. PMID- 17397434 TI - Proxies and consent discussions for dementia research. AB - OBJECTIVES: To better understand the nature of informed consent encounters for research involving patients with dementia that requires proxy consent. DESIGN: Audiotaping of informed-consent encounters for a study of genetic markers for sporadic Alzheimer's disease. SETTING: Outpatients at an Alzheimer's disease research center. PARTICIPANTS: Patients with dementia and their companions. MEASUREMENTS: Audiotapes were analyzed to characterize communication style and coverage of the standard elements of informed consent and, using the Roter Interaction Analysis System, to capture the dynamics of three-way interaction between the patient, their companion, and the physician investigator. RESULTS: Of 26 informed consent encounters, all involved a patient, a companion, and a physician. Patients had a mean Mini-Mental State Examination (MMSE) score of 21.8. For patients, 49% of their interactions involved agreement and approval (positive statements), 16% psychosocial information, 7% biomedical information, 7% asking questions, and 7% expressing emotion. Companion interactions involved 37% positive statements and 19% biomedical information. Physician interactions involved emotional expressiveness (30%) and positive statements (19%). Discussion length was positively related to MMSE score (Spearman rho=0.45; P<.02). Coverage of required elements of informed consent was fairly comprehensive and had no relationship to patients' MMSE scores. CONCLUSION: These data should inform policies regarding the ethically appropriate ways of conducting research with cognitively impaired adults. For example, patients in this study were more silent than their companions and the physician, but when patients spoke, they primarily agreed with what was said. Although this might first seem to signal assent, such an interpretation should be made with caution for persons with dementia. In addition, previous work on informed consent has focused on its cognitive aspects, but these data reveal that the emotional and social dimensions warrant attention. PMID- 17397435 TI - The relationship between the action of arginine vasopressin and responsiveness to oral desmopressin in older men: a pilot study. AB - OBJECTIVES: To identify whether oral desmopressin (ddAVP) reduced nocturnal urine volume (NUV) in older men with nocturia without obvious bladder outlet obstruction and to determine whether deficiencies in arginine vasopressin (AVP) release and action demonstrated using water deprivation testing predicted responsiveness to ddAVP. DESIGN: Participants had a 2-day Clinical Research Center (CRC) evaluation followed by a double-blinded, placebo-controlled, crossover trial of individually titrated oral ddAVP. SETTING: Participants were from a single Department of Veterans Affairs Medical Center. MEASUREMENTS: Maximum urine osmolality and percentage increase in osmolality were measured after subjects received aqueous vasopressin as part of the overnight water deprivation study; these data were used to categorize participants as normal, having partial central AVP deficiency, or having impaired renal responsiveness to AVP. Response to ddAVP was assessed using data from frequency-volume records. RESULTS: Fourteen participants completed the CRC stay and ddAVP trial. Subjects given ddAVP reduced NUV significantly from baseline (P=.02) and had significantly lower NUV than when on placebo (P=.01). The mean net reduction in NUV from ddAVP compared to placebo was 14+/-18%. Using water deprivation testing to categorize participants, 10 were normal, two had partial central AVP deficiency, and two had impaired renal responsiveness. The mean net reduction in NUV for those with abnormal water deprivation tests was 11+/-25%, versus 15+/-16% for those with normal water deprivation testing (P=.70). CONCLUSION: In this small randomized, controlled trial in older men with nocturia, ddAVP reduced NUV. Counter to expectations, participants deemed normal according to water deprivation tests had approximately equivalent responsiveness to ddAVP. Although this study cannot offer definitive conclusions on the lack of prediction of water deprivation testing for ddAVP benefit, these data offer additional information that may help clarify the pathophysiology and optimal treatment of nocturia in older men. PMID- 17397436 TI - Multitasking: association between poorer performance and a history of recurrent falls. AB - OBJECTIVES: To examine the association between poorer performance on concurrent walking and reaction time and recurrent falls. DESIGN: Cross-sectional analysis. SETTING: Community. PARTICIPANTS: Three hundred seventy-seven older community dwelling adults (mean age+/-standard deviation 78+/-3). MEASUREMENTS: Reaction times on push-button and visual-spatial decision tasks were assessed while seated and while walking a 20-m course (straight walk) and a 20-m course with a turn at 10 m (turn walk). Walking times were recorded while walking only and while performing a reaction-time response. Dual-task performance was calculated as the percentage change in task times when done in dual-task versus single-task conditions. A history of recurrent falls (> or = 2 vs < or = 1 falls) in the prior 12 months was self-reported. Multivariate logistic regression models were used to predict the standardized odds ratios (ORs) of recurrent falls history. The standardized unit for dual-task performance ORs was interquartile range/2. RESULTS: On the push-button task during the turn walk, poorer reaction time response (slower) was associated with 28% lower (P=.04) odds of recurrent fall history. On the visual-spatial task, poorer walking-time response (slower) was associated with 34% (P=.02) and 42% (P=.01) higher odds of recurrent falls history on the straight and turn walks, respectively. CONCLUSION: These findings suggest that walking more slowly in response to a visual-spatial decision task may identify individuals at risk for multiple falls. Prospective studies are needed to confirm the prognostic value of poor walking responses in a dual-task setting for multiple falls. PMID- 17397437 TI - The consequences of falls in acute and subacute hospitals in Australia that cause proximal femoral fractures. AB - OBJECTIVES: To compare consequences for patients with proximal femoral fractures (PFFs) sustained in the hospital with patients who sustained PFFs in the community. DESIGN: Data were collected from inpatient notes and incident reports of patients admitted to hospitals over a 6-year period. All patients aged 75 and older sustaining a PFF in the hospital were identified and matched according to sex, age, and fracture date with patients who sustained a PFF in the community. SETTING: Illawarra region hospitals, New South Wales, Australia. PARTICIPANTS: Forty-three patients with a hospital-acquired PFF and 43 patients with a community-acquired PFF; mean age was 84.0 (range 75-92), and 67% were women. MEASUREMENTS: Outcomes at hospital discharge and circumstances of hospital acquired PFF. RESULTS: Comparing outcomes of subjects with hospital versus community-acquired PFF revealed that 12 versus four died in the hospital (P=.03), 14 versus five were discharged to long-term nursing care facilities (P=.02), six versus 18 returned to preadmission ambulation (P=.004), and four versus 24 returned to preadmission activity of daily living status (P<.001). The median postfracture length of stay was 46 days for subjects with hospital-acquired PFF versus 32 days for those sustaining a PFF in the community (P<.01). Review of circumstances of the 43 hospital-acquired fractures revealed that 26 occurred in subacute wards, 25 occurred in bedrooms, 25 occurred at night, and 38 occurred while unsupervised. CONCLUSION: Patients with hospital-acquired PFF have poor outcomes. Adequate supervision, provision of hip protectors in the hospital, and strategies that address fracture circumstances may hold the keys to prevention. PMID- 17397438 TI - Prevalence of peripheral arterial disease and risk factors in persons aged 60 and older: data from the National Health and Nutrition Examination Survey 1999-2004. AB - OBJECTIVES: Peripheral arterial disease (PAD) is associated with significant cardiovascular morbidity and mortality. The study objectives were to examine the prevalence of PAD and associated risk factors. DESIGN: A cross-sectional nationally representative health examination survey. SETTING: The National Health and Nutrition Examination Survey 1999-2004. PARTICIPANTS: Data from 3,947 men and women aged 60 and older who received a lower extremity examination. MEASUREMENTS: The main outcome was PAD, defined as an ankle-brachial blood pressure index of less than 0.9 in either leg. RESULTS: In older U.S. adults, PAD prevalence was 12.2% (95% confidence interval (CI) = 10.9-13.5%). PAD prevalence increased with age. PAD prevalence was 7.0% (95% CI = 5.6-8.4%) for those aged 60 to 69, 12.5% (95% CI = 10.4-14.6%), and 23.2% (95% CI = 19.8-26.7%) for those aged 70 to 79 and 80 and older. Age-adjusted estimates show that non-Hispanic black men and women and Mexican-American women had a higher prevalence of PAD than non-Hispanic white men and women (19.2%, 95% CI = 13.7-24.6%; 19.3%, 95% CI = 13.3-25.2%; and 15.6%, 95% CI = 12.7-18.6%, respectively). The results of the fully adjusted model show that current smoking (OR = 5.48, 95% CI = 3.60-8.35), previous smoking (OR = 1.94, 95% CI = 1.39-2.69), diabetes mellitus (OR = 1.81, 95% CI = 1.12 2.91), low kidney function (OR = 2.69, 95% CI = 1.58-4.56), mildly decreased kidney function (OR = 1.71, 95% CI = 1.22-2.38), high-sensitivity C-reactive protein greater than 3.0 mg/L (OR = 2.69, 95% CI = 1.24-5.85), treated but not controlled hypertension (OR = 1.95, 95% CI = 1.40-2.72), and untreated hypertension (OR = 1.68, 95% CI = 1.13-2.50) were all significantly associated with prevalent PAD. CONCLUSION: PAD prevalence increases with age and is associated with treatable risk factors for cardiovascular disease. PMID- 17397439 TI - Prescribing in palliative care as death approaches. AB - OBJECTIVES: To determine how prescribing for comorbid illnesses and symptom control changes during the palliative phase of a terminal illness. DESIGN: This prospective cohort study explores the relative contribution to prescribing of symptom-specific medications (SSMs) and long-term medications for comorbid medical conditions. SETTING: Regional consultative palliative care program, Adelaide, South Australia. PARTICIPANTS: Two hundred sixty consecutive patients, 96% of whom had cancer, who enrolled and subsequently died in a larger randomized trial exploring palliative service delivery. MEASUREMENTS: Medication and performance data were collected monthly from referral until death (mean 107 days, median 93 days, standard deviation (SD) 103 days, range 11-752 days). Prespecified subgroup analyses of age, performance status, and the baseline use of medications for comorbid medical conditions were performed. RESULTS: At baseline, the mean total number of medications+/-SD was 4.9+/-2.8 (range 0-16), SSMs was 2.3+/-1.5 (range 0-7), and medications for comorbid medical conditions was 2.6+/-2.4 (range 0-13). As death approached, the total number of medications increased because of SSM prescribing (2.5 more medications, 95% confidence interval (CI)=2.2-2.9; P<.001) with a decrease in medications for comorbid medical conditions (1.1 fewer medications, 95% CI=0.8-1.3; P<.001). There was an increase in the number of medications meeting Beers' criteria for high-risk inappropriate medication use for SSMs (29% to 48%). More SSMs were prescribed in people with better performance status, and older participants took more medications for comorbid medical conditions. CONCLUSION: Prescribing changes as life-limiting illnesses progress, with older people taking more medications. Medications for comorbid medical conditions should be reviewed in the context of their original therapeutic goals. PMID- 17397440 TI - Validity of the Patient Health Questionnaire 2 (PHQ-2) in identifying major depression in older people. AB - OBJECTIVES: To determine the performance of the Patient Health Questionnaire 2 (PHQ-2) against the criterion standard for diagnosing major depression, the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), and to examine its performance across age, sex, and racial/ethnic groups. DESIGN: Cross-sectional observational study. SETTING: The National Epidemiologic Survey on Alcohol and Related Conditions (NESARC) (2001/02), a nationally representative survey of the noninstitutionalized U.S. household population. PARTICIPANTS: The 8,205 adults aged 65 and older who participated in NESARC. MEASUREMENTS: The PHQ 2's criterion validity (sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) against the DSM-IV) and construct validity (Spearman correlations between the PHQ-2 and the six scales of the Medical Outcomes Study 12-item Short Form Questionnaire (SF-12)) were calculated. RESULTS: The PHQ-2's criterion validity for major depression was good (sensitivity=100%, specificity=77%, AUC=0.88). Its sensitivity was 100% for each subgroup. Specificity increased with age, was higher for men than for women, and differed across racial and ethnic groups. For the total sample and each sex, all six SF-12 scales were significantly lower in people who tested positive using the PHQ-2. For each age and racial or ethnic group, most of the six scales differed significantly between those who tested positive and tested negative. CONCLUSION: The PHQ-2 is a valid screening tool for major depression in older people but should be followed by a more-comprehensive diagnostic process. Although its specificity differs by age, sex, and racial and ethnic groups, these differences appear to be of little clinical significance. PMID- 17397441 TI - Interventions to reduce fear of falling in community-living older people: a systematic review. AB - The objective was to assess which interventions effectively reduce fear of falling in community-living older people. An extensive search for relevant literature comprised a database search of PubMed, EMBASE, PsycINFO, and the Cochrane Central Register of Controlled Trials; expert consultation; and manually searching reference lists from potentially relevant papers. Randomized, controlled trials that assessed fear of falling in community-living older people were included. Two independent reviewers extracted data from full papers on study characteristics, methodological quality, outcomes, and process characteristics of the intervention. The search identified 599 abstracts, and 19 papers met the inclusion criteria. Seven of those papers were identified using expert consultation. Fifty-five percent of all validity items and 39% of process characteristic items were fulfilled across the 19 trials. Twelve of the 19 papers were of higher methodological quality. In 11 of these trials, fear of falling was lower in the intervention group than in the control group. Interventions that showed effectiveness were fall-related multifactorial programs (n=5), tai chi interventions (n=3), exercise interventions (n=2), and a hip protector intervention (n=1). Three of these interventions explicitly aimed to reduce fear of falling. Several interventions, including interventions not explicitly aimed at fear of falling, resulted in a reduction of fear of falling in community living older people. Limited but fairly consistent findings in trials of higher methodological quality showed that home-based exercise and fall-related multifactorial programs and community-based tai chi delivered in group format have been effective in reducing fear of falling in community-living older people. PMID- 17397442 TI - Attitudes of older Korean Americans toward mental health services. AB - Given the increasing evidence that older ethnic minorities underuse mental health services, the present study assessed determinants of attitudes toward mental health services with a sample of older Korean Americans (N=472). Adapting Andersen's behavioral health model, predisposing factors (age, sex, marital status, education, length of residence in the United States), mental health needs (anxiety, suicidal ideation, depressive symptoms), and enabling factors (personal experiences and beliefs) were considered as potential predictors. Shorter residence in the United States and higher levels of depressive symptoms were associated with more-negative attitudes toward mental health services. Culture influenced personal beliefs (knowledge about mental illness and stigmatism) were found to play a substantial role in shaping individuals' attitudes toward mental health services. Findings call attention to the need to investigate how culture influences the response to mental health needs and to develop community education and outreach programs to close the gaps between mental health needs and service utilization in older ethnic minority populations. PMID- 17397443 TI - Circulating sex hormone levels and aortic stiffness in men. PMID- 17397444 TI - Alzheimer's disease medication: use and cost projections for Medicare Part D. PMID- 17397445 TI - A multidisciplinary management program in primary care to improve hypertension control and healthy behaviors in elderly patients. PMID- 17397447 TI - Patient satisfaction and "the Seven C's". PMID- 17397446 TI - Relationship between hip fracture subtypes and analgesia use. PMID- 17397448 TI - Community-acquired bacterial meningitis in older people. PMID- 17397449 TI - Reversible cognitive and neurological symptoms during valproic acid therapy. PMID- 17397451 TI - Central hypothyroidism allows discovery of a pituitary disorder in older people. PMID- 17397452 TI - Acute increase in blood urea nitrogen caused by enteric nutrition. PMID- 17397453 TI - Validity and reliability of quantitative gait analysis in geriatric patients with and without dementia. PMID- 17397454 TI - The investigation of syncope in older patients. PMID- 17397455 TI - Bathing disability in older patients. PMID- 17397456 TI - Catie's lessons. PMID- 17397458 TI - Whole saliva in X-linked hypohidrotic ectodermal dysplasia. AB - BACKGROUND: X-linked hypohidrotic ectodermal dysplasia (HED) is the most common type of ectodermal dysplasia. Identification of female carriers of X-linked HED can be difficult because of varying degrees of clinical symptoms due to the X chromosome inactivation. This is the first study about whole saliva flow and composition in males affected by HED and female carriers all confirmed by molecular genetic analysis. HYPOTHESIS AND AIM: As salivary glands derive from ectoderm, we hypothesized that whole saliva flow and composition are altered in males affected by HED and female carriers. DESIGN: Saliva flow and composition were examined in a group of affected males and in a group of female carriers, all confirmed by molecular genetic analysis, and compared with healthy male and female controls. RESULTS: Affected males and female carriers had reduced whole saliva flow and saliva with high concentrations of most inorganic salivary constituents as well as total protein. However, affected males and female carriers seemed to have reduced amylase activity and concentration relative to their total protein concentration. CONCLUSION: Saliva flow and composition may be used as part of a comprehensive clinical examination to identify potential female carriers of HED. PMID- 17397459 TI - Dental enamel defects in children with coeliac disease. AB - OBJECTIVE: The aim of this study was to investigate whether Dutch children with proven coeliac disease show specific dental enamel defects, and to asses whether children with the same gastrointestinal complaints, but proved no-coeliac disease, lack these specific dental enamel defects. MATERIALS AND METHODS: Eighty one children (53 coeliac patients and 28 control subjects) were examined during the period 2003-2004 in the Oral Surgery Outpatient Clinic of the Academic Medical Centre in Amsterdam. RESULT: Twenty-nine (55%) coeliac patients had enamel defects against 5 (18%) control subjects. In the coeliac disease group, the enamel defects were diagnosed as specific in 20 (38%) children, compared with 1 (4%) in the control group. Statistical analysis showed significantly more specific enamel defects in children with coeliac disease than in children in the control group (chi(2) = 12.62, d.f. = 2, P = 0.002). CONCLUSION: This study showed significantly more specific enamel defects in Dutch children with coeliac disease as compared with children in the control group. Dentists could play an important role in recognizing patients with coeliac disease. PMID- 17397460 TI - A comparison of paracetamol, ibuprofen or their combination for pain relief following extractions in children under general anaesthesia: a randomized controlled trial. AB - OBJECTIVE: This study was designed to compare the effectiveness of different oral analgesics for relieving pain and distress in children following the extraction of teeth under general anaesthesia (GA). The analgesics included paracetamol alone, ibuprofen alone, and paracetamol and ibuprofen in combination. METHODS: Two hundred and one subjects were randomly allocated to one of four groups. Forty seven children were included in the ibuprofen alone (5 mg kg(-1)) group, 51 in the paracetamol/ibuprofen combination (15/5 mg kg(-1)) group, 48 in the high-dose paracetamol (20 mg kg(-1)) group, and 55 children were included in the usual-dose paracetamol (15 mg kg(-1)) group (control group). Evaluation of distress for children was made immediately pre-operatively, on recovery from anaesthesia and again after 15 min by using a five-point face scale. Furthermore, each child was observed immediately postoperatively and 15 min postoperatively for signs of pain using the Children's Hospital of Eastern Ontario Pain Scale. RESULTS: There were significant decreases in the mean pain and distress scores for both the ibuprofen alone and paracetamol/ibuprofen combination groups compared to the control group (usual-dose paracetamol) at 15 min postoperatively. CONCLUSIONS: This study provides evidence to support the oral administration of ibuprofen alone or in combination with paracetamol for postoperative analgesia in children who are having teeth extracted under GA. PMID- 17397461 TI - The relationship between a primary maxillary incisor with a talon cusp and the permanent successor: a study of 57 cases. AB - BACKGROUND: Most reported cases of talon cusps in the primary dentition have been on the maxillary central incisors and the permanent successors have remained unaffected. Four of the five reported cases on a maxillary lateral incisor, however, have been associated with a supernumerary permanent successor. AIM: This paper describes the relationship between the presence of talon cusps on a primary maxillary incisor, and the morphology and number of the permanent successors in a population of Chinese children. DESIGN: The dental records of children diagnosed with a talon cusp on a primary incisor were retrieved for review. The diagnoses took place in a regional school dental clinic in Hong Kong between April 2002 and August 2005. RESULTS: Fifty-eight primary maxillary incisors with talon cusps were found. When the central incisors were involved, 32 of the 35 (91.4%) underlying permanent successors were not found to be associated with any odontogenic abnormalities. When the lateral incisors were involved, however, 18 of the 23 cases (78.3%) showed odontogenic abnormalities, 14 of which were associated with supernumerary teeth. CONCLUSIONS: The present study shows that, when there is a talon cusp on a primary maxillary lateral incisor, a high proportion of the underlying permanent successors can be expected to exhibit odontogenic abnormalities. PMID- 17397462 TI - Vital pulpotomy in the primary dentition: attitudes and practices of community dental staff in Wales. AB - OBJECTIVE: The International Agency for Research on Cancer (IARC) has classified formaldehyde as carcinogenic to humans, leaving the dental profession to look for viable substitutes to formocresol in the vital pulpotomy technique. This study was designed to examine the attitudes and practices of Community Dental Service (CDS) staff in Wales in relation to vital pulpotomy for primary molars 18 months following the IARC's press release. METHODS: The study employed a postal questionnaire. RESULTS: Questionnaires were returned by 79 (78.2%) of the CDS staff surveyed, yielding a sample of 65 dentists practising the technique. The most commonly used pulpotomy agents were formocresol, paraformaldehyde and ferric sulphate. Twenty-seven (41.5%) dentists expressed concern regarding their preferred pulpotomy agent and 17 (26.2%) were considering changing their technique. Only one respondent (1.5% of the sample) routinely took preoperative radiographs; follow-up radiographs were routinely taken by only three dentists (4.6%). Only 44 respondents (67.7%) always used local anaesthesia for this form of treatment. Amalgam was the most commonly used restorative material. Twenty-two respondents (33.8%) stated that they would pulp treat a primary molar on more than one occasion. CONCLUSIONS: The results of this study suggest that there is need for relevant continuing professional development courses for CDS staff in Wales. PMID- 17397463 TI - Prevalence of four putative periodontopathic bacteria in saliva of a group of Brazilian children with mixed dentition: 1-year longitudinal study. AB - AIM: This study aimed at evaluating the prevalence of putative periodontal pathogens (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella nigrescens, Treponema denticola) in saliva of children with mixed dentition at two different time points, correlating these findings with a clinical parameter of gingival health. DESIGN: Polymerase chain reaction (PCR) detection was used to determine the prevalence of these bacteria in saliva of 64 children in 2003 and 60 children in 2004. Gingival health was assessed by gingival index. RESULT: Sixty-two (96.9%) and 50 (83.3%) children presented low gingival inflammation, whereas 2 (3.1%) and 10 (16.7%) had moderate scores in 2003 and 2004, respectively. Majority of the children (81.3% in 2003 and 73.3% in 2004) had detectable levels of at least one of the bacteria. The prevalence found was of 4.7% and 1.7% for A. actinomycetemcomitans, 6.3% and 8.3% for P. gingivalis, 23.4% and 48.3% for P. nigrescens (P < 0.05), and 71.9% and 50% for T. denticola (P < 0.05) in 2003 and 2004, respectively. No significant relationship between gingival index and presence of these bacteria and combination of different species was found. CONCLUSION: A high percentage of children harboured at least one of the putative periodontal pathogens in saliva, but presented periodontally healthy conditions. PMID- 17397464 TI - The accompanying adult: authority to give consent in the UK. AB - BACKGROUND: Children may be accompanied by various people when attending for dental treatment. Before treatment is started, there is a legal requirement that the operator obtain informed consent for the proposed procedure. In the case of minors, the person authorized to give consent (parental responsibility) is usually a parent. AIM: To ascertain if accompanying persons of children attending the Department of Paediatric Dentistry at the Eastman Dental Hospital, London were empowered to give consent for the child's dental treatment. DESIGN: A total of 250 accompanying persons of children attending were selected, over a 6-month period. A questionnaire was used to establish whether the accompanying person(s) were authorized to give consent. RESULT: The study showed that 12% of accompanying persons had no legal authority to give consent for the child's dental treatment. CONCLUSION: Clinicians need to be aware of the status of persons accompanying children to ensure valid consent is obtained. PMID- 17397465 TI - Perception of dental fluorosis and other oral health disorders by 12-year-old Brazilian children. AB - AIM: The aim of this study was to verify the perception of dental fluorosis as an oral health problem by 12-year-old Brazilian children and to evaluate if they were able to detect other oral disorders. METHODOLOGY: The sample consisted of 401 schoolchildren. Firstly, clinical examinations were performed using a visual method under natural light. After that, children answered a questionnaire with the purpose of assessing the self-perception of their oral health problems. Next, the volunteers were shown a photo album containing 24 photographs, and had to match each photo with a severity scale. RESULTS: The prevalence of fluorosis was 18.2%; 81.8% of this total in fluorosis scale T-F 1. Of the 401 children, 48.9% (n = 196) answered oral health problems related to concerns, such as aesthetic appearance or pain. Among them, only two children, both with fluorosis T-F 2, correlated their problems with the presence of stained teeth. As regards the photo album analysis, the children considered photos showing fluorosis T-F 7-9 as the most severe, whereas the photo of an orally healthy patient was considered the least severe. CONCLUSION: Children did not show negative perception of dental fluorosis, except for dental fluorosis at severe levels (T-F 7-9), and were mainly able to detect aesthetic or pain-related problems. PMID- 17397466 TI - Langerhans cell histiocytosis in a child: a 10-year follow-up. AB - BACKGROUND: This report describes a case of the benign form of Langerhans cell histiocytosis, which involves only the maxilla and mandible, and has no extraskeletal manifestations. CASE REPORT: A 2.9-year-old boy was referred to a paediatric dentistry clinic, where clinical, radiographic and histological findings were found to be consistent with Langerhans cell histiocytosis. The difficulties involved in the diagnosis and the subject's functional oral rehabilitation are described. In view of the presence of large bone lesions, systematic treatment consisted of oral chemotherapy and the administration of methylprednisolone by intralesional infiltration. Oral examination revealed complete resolution of the ulcerative lesions. The soft tissue appeared healthy and had a normal colour. There was no tooth mobility and radiography provided evidence of bony healing. CONCLUSION: This case is particularly interesting because follow-up clinical and radiographic observations have been made over the past 10 years. The successful treatment had several objectives, including providing the patient with an aesthetic appearance and recovering the lost vertical dimension of occlusion. PMID- 17397467 TI - Development of acute Henoch-Schonlein purpura subsequent to endodontic treatment. AB - BACKGROUND: Henoch-Schonlein purpura (HSP) is an inflammatory disorder of unknown origin that is characterized by IgA-dominant immune complexes in smaller blood vessels. It results in a triad of symptoms, including a purpuric rash on the lower extremities, abdominal pain or renal involvement, and arthritis. Any of the triad may be absent, however, which often leads to confusion in diagnosing the condition. Cases of acute HSP developing subsequent to dental treatment have not been previously reported in the dental literature. CASE REPORT: This study reports the unusual case of a 14-year-old female who developed acute HSP following endodontic treatment. CONCLUSION: Treatment for this condition is supportive and children affected by this disorder need close follow-up of their renal status. PMID- 17397468 TI - Plasmacytoid myoepithelioma of the palate in a child. AB - BACKGROUND: Myoepithelioma is a rare benign salivary gland tumour, localized most frequently in the parotid and in minor intraoral salivary glands. There have been only four cases of myoepithelioma in children and adolescents reported in the English-language literature, all of them involving the plasmacytoid variant. CASE REPORT: A 13-year-old boy, complained of a painless nodule of the palate. Incisional biopsy was performed and revealed large plasmacytoid cells with a round and eccentric nuclei. The diagnosis was consistent with myoepithelioma, plasmocytoid variant. Tumour cells were positive for cytokeratins, vimentin and S 100 protein. Surgical resection was performed and no evidences of tumour recurrence were observed after 6 years of the treatment. CONCLUSION: Myoepithelioma is a very rare tumour in children and apparently presents a good prognosis, similar to occur in adult patients. PMID- 17397469 TI - Subcutaneous emphysema during restorative dentistry. AB - BACKGROUND: Subcutaneous emphysema is a rare occurrence in the dental setting. When it does occur, the entity may be mistaken for an anaphylactic reaction to a local anaesthetic agent or other medications used in dental surgery. CASE REPORT: During restorative treatment of a left second primary molar air was introduced under an operculum. Communication with submandibular and sublingual spaces to the mediastinum was established. CONCLUSION: Therapy for subcutaneous emphysema involves, correct diagnosis and antibiotic coverage. Subcutaneous air will resolve over time so observation is indicated. PMID- 17397470 TI - Cultivable bacterial diversity from the human colon. AB - Knowledge of the composition of the colonic microbiota is important for our understanding of how the balance of these microbes is influenced by diet and the environment, and which bacterial groups are important in maintaining gut health or promoting disease. Molecular methodologies have advanced our understanding of the composition and diversity of the colonic microbiota. Importantly, however, it is the continued isolation of bacterial representatives of key groups that offers the best opportunity to conduct detailed metabolic and functional studies. This also permits bacterial genome sequencing which will accelerate the linkage to functionality. Obtaining new human colonic bacterial isolates can be challenging, because most of these are strict anaerobes and many have rather exact nutritional and physical requirements. Despite this many new species are being isolated and described that occupy distinct niches in the colonic microbial community. This review focuses on these under-studied yet important gut anaerobes. PMID- 17397471 TI - A quantitative real-time PCR assay for the highly sensitive and specific detection of human faecal influence in spring water from a large alpine catchment area. AB - AIMS: The aim of the study was the development of a sensitive human-specific quantitative real-time PCR assay for microbial faecal source tracking (MST) in alpine spring water. The assay detects human-specific faecal DNA markers (BacH) from 16S rRNA gene sequences from the phylum Bacteroidetes using TaqMan minor groove binder probes. METHODS AND RESULTS: The qualitative and quantitative detection limits of the PCR assay were 6 and 30 marker copies, respectively. Specificity was proved by testing 41 human faeces and waste water samples and excluding cross-amplification from 302 animal faecal samples from Eastern Austria. Marker concentrations in human faecal material were in the range from 6.6 x 10(9) to 9.1 x 10(10) marker equivalents per gram. The method was sensitive enough to detect a few 100 pg of faeces in faecal suspensions. The assay was applied on water samples from an alpine karstic spring catchment area and the results reflected the expected levels of human faecal influence. CONCLUSIONS: The method exhibited sufficient sensitivity to allow quantitative source tracking of human faecal impact in the investigated karstic spring water. SIGNIFICANCE AND IMPACT OF THE STUDY: The developed method constitutes the first quantitative human-specific MST tool sensitive enough for investigations in ground and spring water. PMID- 17397472 TI - Thermal death of Escherichia coli O157:H7 in cattle feeds. AB - AIMS: To determine if the temperatures used in feed manufacture are likely to destroy Escherichia coli O157. METHODS AND RESULTS: Two commercial feeds were ground and inoculated with E. coli O157 cells. The feeds were heated to 50, 55, 60, 65 or 70 degrees C. Heating produced quadratic survivor curves, with rapid initial decreases. The survival characteristics of E. coli O157 differed in the two feeds. The reductions observed in one feed may not have been due to heat alone. There was evidence that indigenous anti-E. coli O157 factor(s) in one feed acted with the heat and contributed to the observed rates of bacterial death. Heating at 70 degrees C for 20 or 120 s resulted in approx. 1.3 and 2.2 log reductions in E. coli O157 numbers respectively. Lesser reductions were observed at lower temperatures. CONCLUSIONS: The time/temperature combinations used in commercial pelleting processes would not effectively kill high numbers of E. coli O157. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to look at the survival of E. coli O157 strains after heat treatment within concentrated animal feed. The study provides information on the likely risk of E. coli O157 surviving the animal feed manufacturing process. PMID- 17397473 TI - Isolation of polyhydroxyalkanoates-producing bacteria using a combination of phenotypic and genotypic approach. AB - AIMS: To develop an efficient approach using a combination of phenotypic and genotypic methods for isolation of environmental bacteria that produce mid-chain length polyhydroxyalkanoates (mcl-PHAs). METHODS AND RESULTS: A viable-colony staining method using Nile red was used to screen for PHA-producing bacteria followed by a polymerase chain reaction (PCR) screen using primers to amplify the partial nucleic acid sequence of the phaC1 synthase gene for confirmation. Microbes containing lipophilic storage compounds isolated from environmental samples could readily be detected by the colony staining method. They were further examined by Sudan Black staining to highlight the inclusions inside the cells. These isolates were subsequently subjected to PCR analysis. As a result, more than a hundred strains were identified as PHA-positive isolates from this screening approach. CONCLUSIONS: These results conclusively demonstrate that environmental bacterial strains able to accumulate the PHAs could readily be obtained by this screening method. SIGNIFICANCE AND IMPACT OF THE STUDY: We propose a polyphasic approach using a combination of phenotypic and genotypic screening method to rapidly screen and identify bacteria able to produce significant amounts of mcl-PHAs from environment. This approach can be adopted as a rapid screen for micro-organisms able to accumulate PHAs to be used for potential manufacture and other industrial applications. PMID- 17397474 TI - A marine bacterial adhesion microplate test using the DAPI fluorescent dye: a new method to screen antifouling agents. AB - AIMS: To develop a method to screen antifouling agents against marine bacterial adhesion as a sensitive, rapid and quantitative microplate fluorescent test. METHODS AND RESULTS: Our experimental method is based on a natural biofilm formed by mono-incubation of the marine bacterium Pseudoalteromonas sp. D41 in sterile natural sea water in a 96-well polystyrene microplate. The 4'6-diamidino-2 phenylindole dye was used to quantify adhered bacteria in each well. The total measured fluorescence in the wells was correlated with the amount of bacteria showing a detection limit of one bacterium per 5 microm(2) and quantifying 2 x 10(7) to 2 x 10(8) bacteria adhered per cm(2). The antifouling properties of three commercial surface-active agents and chlorine were tested by this method in the prevention of adhesion and also in the detachment of already adhered bacteria. The marine bacterial adhesion inhibition rate depending on the agent concentration showed a sigmoid shaped dose-response curve. CONCLUSIONS: This test is well adapted for a rapid and quantitative first screening of antifouling agents directly in seawater in the early steps of marine biofilm formation. SIGNIFICANCE AND IMPACT OF THE STUDY: In contrast to the usual screenings of antifouling products which detect a bactericidal activity, this test is more appropriate to screen antifouling agents for bacterial adhesion removal or bacterial adhesion inhibition activities. This screening test focuses on the antifouling properties of the products, especially the initial steps of marine biofilm formation. PMID- 17397475 TI - Environmental microbial contamination in a stem cell bank. AB - AIM: The aim of this study was to evaluate the main environmental microbial contaminants of the clean rooms in our stem cell bank. METHODS AND RESULTS: We have measured the microbial air contamination by both passive and active air sampling and the microbial monitoring of surfaces by means of Rodac plates. The environmental monitoring tests were carried out in accordance with the guidelines of European Pharmacopeia and US Pharmacopeia. The micro-organisms were identified by means of an automated system (VITEK 2). During the monitoring, the clean rooms are continually under good manufacturing practices specifications. The most frequent contaminants were Gram-positive cocci. CONCLUSIONS: The main contaminants in our stem cell bank were coagulase-negative staphylococci and other opportunistic human pathogens. In order to assure the levels of potential contamination in both embryonic and adult stem cell lines, a continuous sampling of air particles and testing for viable microbiological contamination is necessary. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first evaluation of the environmental contaminants in stem cell banks and can serve as initial evaluation for these establishments. The introduction of environmental monitoring programmes in the processing of stem cell lines could diminish the risk of contamination in stem cell cultures. PMID- 17397476 TI - Production of a COX-2 inhibitor, 2,4,5-trimethoxybenzaldehyde, with submerged cultured Antrodia camphorata. AB - AIMS: To investigate the active ingredient in fruiting bodies and to produce it with cultured mycelium in Antrodia camphorata (BCRC 35398). METHODS AND RESULTS: The volatile components from the fruiting bodies, the liquid cultured broth of A. camphorata and Cinnamomum kanehirae wood were separately isolated by steam distillation-solvent extraction and identified by gas chromatography-mass spectrometry. In the fruiting bodies, a COX-2 inhibitor 2,4,5 trimethoxybenzaldehyde (TMBA) was found to be the most abundant constituent, but was totally absent in its cultured broth and its natural host, C. kanehirae wood. On feeding with the acid-digested sawdust of C. kanehirae wood or vanillin to the broth for culture, TMBA was produced in both cultured broths. CONCLUSION: The TMBA identified in fruiting bodies was an active ingredient whose functions consisted with the reported experiences of this mushroom. Feeding vanillin to culture broth could produce TMBA containing mycelium product like its fruiting bodies did. SIGNIFICANCE AND IMPACT OF THE STUDY: This study found an active ingredient in fruiting bodies of A. camphorata and elucidated this compound derived from digested sawdust of C. kanehirae wood. A feasible method was also developed to produce TMBA containing mycelium by feeding vanillin. PMID- 17397477 TI - Improving energy accumulation of microbial fuel cells by metabolism regulation using Rhodoferax ferrireducens as biocatalyst. AB - AIMS: To study the physiology and metabolism of microbial cells in the performance of microbial fuel cells (MFCs). METHODS AND RESULTS: A dual-chamber MFCs was constructed, and Rhodoferax ferrireducens was used as biocatalyst. To examine the physiology of microbial cells in the performance of MFCs, the anode media containing planktonic cells was replaced with fresh media in which KH(2)PO(4) and/or NH(4)Cl were excluded. The replacing of anode media containing planktonic cells with fresh media excluded of KH(2)PO(4) and NH(4)Cl made the coulombic yield remarkably increased by a factor of 68% (from 29.1 to 46.8C). The results showed that the electricity could be generated with cells in biofilms as biocatalyst, and coulombic yield was improved by limiting cell growth via removal of ingredients in anode media. By supplementation of glucose to the anode media when current declined to baseline, MFCs achieved about same platform current values immediately. MFCs could continue to produce electricity for about 30 h even after glucose was below detection. CONCLUSIONS: Biofilms and metabolism of glucose play important roles in the performance of MFCs. Coulombic yield of MFCs could be improved by regulating the media ingredients using the stable biofilms electrode system. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first attempt to study the effect of ingredient compositions of anode media on the performance of MFCs. The observed results that MFCs continued to produce electricity after glucose was below detection was helpful to better understand the mechanism of microbial electricity production. PMID- 17397478 TI - Eco-physiological characterization of fluorescence in situ hybridization probe targeted denitrifiers in activated sludge using culture-independent methods. AB - AIMS: This study proposes the application of a culture-independent method [fluorescence in situ hybridization (FISH)] and a bioreactor operation control strategy to characterize environmental micro-organisms according to their survival strategies in a mixed suspension culture. Eco-physiological characteristics of two 16S rRNA probe-targeted denitrifiers (DEN581 and DEN124) were investigated against the availability of two resources. METHODS AND RESULTS: Four sequencing batch reactors were operated with manipulation of the sludge retention times to enforce limited and excess availability of two nutrients, namely acetate and nitrite, to the biomass. DEN581 FISH probe-targeted denitrifiers demonstrated dominance when the ratio of either acetate or nitrite to biomass was in excess, while DEN124-targeted organisms dominated when the above were limited. CONCLUSIONS: The study demonstrated that microbial populations in mixed cultures can be selected by changing the substrate availability (Rs) to biomass (X) ratio. The manipulation of the specific resource availability (Rs/X) determined which one of the studied probe-targeted denitrifiers (DEN124 or DEN581) became dominant. Rs/X provides a basis to study the physiology of micro-organisms that cannot be isolated in pure culture from activated sludge. SIGNIFICANCE AND IMPACT OF THE STUDY: The eco-physiological characterization of micro-organisms responsible for biological nutrient removal is anticipated to assist process designers and operators to optimize a specific biological process, such as denitrification. PMID- 17397479 TI - Prevalence and growth of Listeria monocytogenes in naturally contaminated cold smoked salmon. AB - AIM: To investigate Listeria monocytogenes contamination and behaviour in naturally contaminated French cold-smoked salmon (CSS). METHOD AND RESULTS: Between 2001 and 2004, L. monocytogenes was detected in 104 of 1010 CSS packs, produced by nine French plants, with different prevalence (from 0% to 41%). The initial contamination, measured with a sensitive filtration method, was low (92% of contaminated products below 1 CFU g(-1)) and growth was limited. CONCLUSION: Growth was consistent with results of a predictive model including microbial competition. SIGNIFICANCE AND IMPACT OF THE STUDY: To be included in a quantitative risk assessment. PMID- 17397480 TI - Strains of Mesorhizobium amorphae and Mesorhizobium tianshanense, carrying symbiotic genes of common chickpea endosymbiotic species, constitute a novel biovar (ciceri) capable of nodulating Cicer arietinum. AB - AIMS: To identify several strains of Mesorhizobium amorphae and Mesorhizobium tianshanense nodulating Cicer arietinum in Spain and Portugal, and to study the symbiotic genes carried by these strains. METHODS AND RESULTS: The sequences of 16S-23S intergenic spacer (ITS), 16S rRNA gene and symbiotic genes nodC and nifH were analysed. According to their 16S rRNA gene and ITS sequences, the strains from this study were identified as M. amorphae and M. tianshanense. The type strains of these species were isolated in China from Glycyrrhiza pallidiflora and Amorpha fruticosa nodules, respectively, and are not capable of nodulating chickpea. These strains carry symbiotic genes, phylogenetically divergent from those of the chickpea isolates, whose nodC and nifH genes showed more than 99% similarity with respect to those from Mesorhizobium ciceri and Mesorhizobium mediterraneum, the two common chickpea nodulating species in Spain and Portugal. CONCLUSIONS: The results from this study showed that different symbiotic genes have been acquired by strains from the same species during their coevolution with different legumes in distinct geographical locations. SIGNIFICANCE AND IMPACT OF THE STUDY: A new infrasubspecific division named biovar ciceri is proposed within M. amorphae and M. tianshanense to include the strains able to effectively nodulate Cicer arietinum. PMID- 17397481 TI - Virulence markers and serotypes of Shiga toxin-producing Escherichia coli, isolated from cattle in Rio Grande do Sul, Brazil. AB - AIMS: To determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) and serotypes and virulence markers of the STEC isolates from beef and dairy cattle in Rio Grande do Sul, Brazil. METHODS AND RESULTS: Faecal samples from beef cattle were collected at slaughterhouses. The isolates were submitted to colony hybridization assay with specific DNA probes for stx1, stx2 and eae genes, and serotyped for the identification of O and H antigens. Thirty-nine per cent of beef cattle surveyed harboured at least one STEC strain. Among the distinct serotypes identified, 10 were shared by both beef and dairy cattle. Most of the strains isolated harboured stx2. Genotypic and phenotypic profiles allowed the identification of 34 and 31 STEC strains, isolated from beef and dairy cattle, respectively. Serotypes O10:H14, O15:H21, O96:H21, O119:H4, O124:H11, O128:H21, O137:H-, O141:H19, O159:H42, O160:H2 and O177:H11, identified in this study, have not been previously reported as STEC isolated from cattle. CONCLUSIONS: Cattle are an important reservoir of STEC strains associated with human diseases in South America. SIGNIFICANCE AND IMPACT OF THE STUDY: Determining the prevalence, genotypic profile and serotypes of STEC strains isolated from cattle enables the prediction of possible risk for public health. PMID- 17397482 TI - Surface properties of Vibrio vulnificus. AB - AIMS: Vibrio vulnificus adheres to a diverse range of surfaces, ranging from the chitinous exoskeleton of mollusks to human tissue. To determine whether environmental and human clinical isolates exhibit different adhesion traits, we studied the ability of 10 environmental isolates and 10 clinical isolates to adhere to human epithelial cells and hydrocarbons with log P values ranging from 3.1 to 8.2. METHODS AND RESULTS: All isolates adhered to varying levels to epithelial cells, and were inhibited to various extents from adherence by mannose and fructose. There was a lack of correlation between adherence to either hydrocarbons or cells and colony opacity. Adherence to hydrocarbons was optimal for solvents with a log P < 8.2. CONCLUSIONS: Vibrio vulnificus clinical and environmental isolates exhibit differential adherence to epithelial cells and hydrocarbons. SIGNIFICANCE AND IMPACT OF THE STUDY: The differential adherence of organisms to hydrocarbons based on log P may have utility in drug design and enhancement of food safety. PMID- 17397483 TI - Comparison of the VIDAS system, FTA filter-based PCR and culture on SM ID for detecting Salmonella in Dermanyssus gallinae. AB - AIMS: To compare different analytical methods for detecting Salmonella in Dermanyssus gallinae. METHODS AND RESULTS: The detection limit of three Salmonella detection methods [Vitek immunodiagnostic assay (VIDAS) Salmonella immuno-concentration/immunoassay, FTA filter-based PCR, and Salmonella detection and identification medium (SM ID) preceded by a pre-enrichment step] was evaluated by crushing mites in serial dilutions of pure cultures of Salmonella enterica ssp. Enterica serotype Enteritidis. Each method was then compared for its ability to detect Salmonella in artificially contaminated mites. In 105 mites artificially engorged with Salm. Enteritidis-contaminated blood, Salmonella was isolated from 68 samples of the samples cultured on SM ID and tests were positive for Salmonella using FTA filter-based PCR and VIDAS in 77 and 65 samples, respectively. Using SM ID as our reference method, specificities and sensitivities were 97% and 94% and 73% and 98.5% for VIDAS and PCR, respectively. CONCLUSIONS: Each method allowed the detection of Salmonella in contaminated mites and is usable for screening mites. PCR is more sensitive but less specific than VIDAS for detecting Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that the VIDAS has been used to detect pathogens in vectors. The development of analytical methods for Salmonella detection in mites is a necessary step in the study of the role of D. gallinae as a vector of salmonellae and to check the contamination of D. gallinae in poultry facilities. PMID- 17397484 TI - Effect of intracellular trehalose in Cryptococcus laurentii and exogenous lyoprotectants on its viability and biocontrol efficacy on Penicillium expansum in apple fruit. AB - AIMS: To improve viability and biocontrol efficacy of Cryptococcus laurentii after freeze drying and in subsequent storage. METHODS AND RESULTS: Viability of C. laurentii was improved after freeze drying and in subsequent storage at 4 or 25 degrees C by using skimmed milk (SM) and sugars (glucose, galactose, sucrose and trehalose) as protectants. Sugars and SM mixed together showed better protection than when they were used separately. Citric acid used as carbon source could induce accumulation of intracellular trehalose in the yeast. The yeast cells with high trehalose level (HT cells) had higher viability than those with low trehalose level (LT cells) after freeze drying and storage for 90 days. After storage for 90 days at 4 degrees C, the HT cells plus SM and sugars as protectant showed a similar biocontrol effect against blue mould rot in apple fruit caused by Penicillium expansum as fresh cells. CONCLUSIONS: Increasing intracellular trehalose content of C. laurentii and adding exogenous protectant (sugars + SM) could improve its viability and maintain its biocontrol efficacy. SIGNIFICANCE AND IMPACT OF THE STUDY: The results have a potential value for commercial application of C. laurentii. PMID- 17397485 TI - Rapid identification of Enterococcus italicus by PCR with primers targeted to 16S rRNA gene. AB - AIMS: To develop a species-specific PCR assay with primers targeted to 16S rRNA gene for the identification of Enterococcus italicus, a new species of Enterococcus, involved in the production of Italian cheeses. METHODS AND RESULTS: The type strain of E. italicus (DSM 15952(T) - 16S rRNA gene accession no. AJ582753) and other strains of the species were subjected to a rapid identification by PCR using primer pairs located within the 16S rRNA gene. A species-specific PCR product of approximately 323 bp was obtained after amplification of all E. italicus strains tested. The specificity of the primers was validated with representatives of the most closely related genera and species and a number of other bacterial species. In addition, the technique enabled the recognition of E. italicus from cheeses. CONCLUSIONS: The protocol was highly efficient and sensitive, enabling the identification of E. italicus from cheeses. SIGNIFICANCE AND IMPACT OF THE STUDY: The species-specific PCR offers a reliable and rapid alternative to conventional phenotypic methods for the identification of E. italicus within the heterogeneous genus Enterococcus. PMID- 17397486 TI - Temperature-dependent biotransformation of 2,4'-dichlorobiphenyl by psychrotolerant Hydrogenophaga strain IA3-A: higher temperatures prevent excess accumulation of problematic meta-cleavage products. AB - AIMS: The present work investigates the possibility that temperature could regulate the pattern of transformation of 2,4'-chlorobiphenyl (2,4'-CB) by psychrotolerant Hydrogenophaga sp. IA3-A. METHODS AND RESULTS: Transformation of 2,4'-chlorobiphenyl to 2- and 4-chlorobenzoic acid (2- and 4-CBA), and meta cleavage products by cells of strain IA3-A incubated at 10 degrees C, 25 degrees C, 37 degrees C or 45 degrees C were monitored by UV spectrometry, HPLC and GC-MS analyses. Cultures incubated at 10 degrees C, 25 degrees C or 37 degrees C produced low amounts of CBAs and excess levels of meta-cleavage products from 2,4'-CB. Cultures incubated at 45 degrees C transformed most of the degraded 2,4' CB to CBAs and low level of meta-cleavage product. Culture extracts contained unusual varieties of isomeric hydroxylated metabolic products. CONCLUSIONS: Efficient transformation of 2,4'-CB to CBAs was possible in cultures incubated at 45 degrees C. Evidence for the involvement of multiple pathways in the transformation of 2,4'-CB in strain IA3-A suggests that differential regulation of the pathways at different temperatures was likely responsible for the change in the pattern of transformation of 2,4'-CB in cultures incubated at 45 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: It may be possible to condition cells to transform chlorinated biphenyls more efficiently without accumulating excess level of toxic intermediates. PMID- 17397487 TI - Observations of cell cluster hollowing in Staphylococcus epidermidis biofilms. PMID- 17397490 TI - European guidelines for quality assurance in cervical cancer screening: recommendations for cytology laboratories. AB - The quality of a cervical cytology laboratory depends on adequate handling and staining of the samples, screening and interpretation of the slides and reporting of the results. These guidelines give an overview of procedures recommended in Europe to manage the balance between best patient care possible, laboratory quality assurance and cost effectiveness and will be published as a chapter 4 in the European Guidelines for Quality Assurance in Cervical Cancer Screening. The laboratory guidelines include protocols for personnel and organisation, material requirements, handling and analysing cervical samples, recording of results, quality management and communication. The section on quality management is comprehensive and includes protocols for all aspects of internal and external quality assurance. The guidelines are extensively referenced and as far as possible the recommendations are evidence-based. PMID- 17397491 TI - Is urinary tract cytology still useful for diagnosis of bladder carcinomas? A large series of 592 bladder washings using a five-category classification of different cytological diagnoses. AB - BACKGROUND: The aim of this study was to estimate the efficiency of a recent five category urinary cytological classification. METHODS: A total of 592 bladder washings were fixed immediately with Saccomanno's fixative. All samples were centrifuged in a Hettich cyto-centrifuge. For each sample, the reference standard was the histology when a lesion was present at the time of cystoscopy. A five category cytological classification was used: negative, suspicious of low (S-Lg) or high (S-Hg) grade neoplasia and consistent with low (Lg) or high (Hg) grade neoplasia. RESULTS: For cytological diagnoses of S-Lg and Lg, sensitivity was 37% and specificity was 95% for the histological diagnosis of low-grade non-invasive urothelial papillary tumour (Lg-UPT), which included papillary urothelial neoplasm of low malignant potential and low-grade urothelial carcinoma. For cytological diagnosis of S-Hg and Hg, sensitivity was 44% for high-grade non invasive urothelial papillary carcinoma (Hg-UPC), 70% for carcinoma in situ (CIS) and 81% for invasive carcinoma (T1 and higher). Specificity was 99% in each case. Cytological diagnosis of S-Hg or Hg was not found for Lg-UPT (0/59) and no cytological diagnosis of S-Lg or Lg was found for invasive carcinoma, but was seen for Hg-UPC in 10% (3/28) and for CIS in 6% (3/50) of cases. CONCLUSION: Despite the absence of international consensus, the recent five-category cytological classification for urine is accurate for current urological practice. PMID- 17397492 TI - Expression of cytokeratin 20 in urine cytology smears: a potential marker for the detection of urothelial carcinoma. AB - BACKGROUND: Urine cytomorphology is one of the oldest methods for screening and monitoring patients with transitional cell carcinoma (TCC). Sensitivity of urine cytology is relatively low. Ancillary techniques on urine sample may increase the sensitivity. AIM: To explore the utility of cytokeratin 20 (CK20) immunostaining in identifying malignant cells in urine cytology smears. MATERIALS AND METHODS: Fourteen cases each of confirmed TCC and benign urinary cytology along with five cases of atypical cells in urine were immunostained with a monoclonal CK20 antibody. Of 14 cases of TCC, 12 showed strong positive staining with the antibody. All benign cases were negative except for a few cases in which the umbrella cells were weakly to moderately positive. In all five cases of atypical urine cytology the atypical cells stained positive with the antibody. These cases were later confirmed as TCC on histopathology of bladder wall biopsy. CONCLUSION: CK20 is an important biomarker that can be used to identify TCC in urine cytology smears. It is particularly useful in those cases where malignancy cannot be confirmed by morphology alone. PMID- 17397493 TI - Detection of urinary bladder cancer with flow cytometry and hexaminolevulinate in urine samples. AB - OBJECTIVES: Urinary bladder urothelial carcinoma is diagnosed by a combination of cystoscopy and biopsy, with cytology as a valuable additional technique. The accuracy of cytological diagnosis depends on the experience of the cytologist and can inevitably vary from one cytologist to another. There is a need for an easy, reliable and objective diagnostic method. In the present study a new method was designed for the detection of bladder cancer cells in urine. METHODS: Flow cytometry was utilized to detect protoporphyrin IX in an artificial model consisting of normal urinary bladder transitional epithelial cells (NBECs) from healthy volunteers' urine and an established human urinary bladder carcinoma cell line, TCCSUP, after incubation with hexaminolevulinate (HAL). In addition, urine samples from 19 patients with histopathologically confirmed superficial bladder cancer were examined. RESULTS: Incubation of NBECs or TCCSUP cells with HAL for 1 hour resulted in production of protoporphyrin IX only in the TCCSUP cells. Incubation of a mixture of NBECs and TCCSUP cells with HAL gave rise to a separated subpopulation of cells with protoporphyrin IX fluorescence. After cell sorting by flow cytometry the protoporphyrin IX-containing subpopulation of cells was confirmed as TCCSUP cells on cytological examination. It was possible to detect 5% TCCSUP cells in the mixture of NBECs/TCCSUP cells. To test the feasibility of the method in clinica diagnosis, urine samples from patients with bladder cancer were also measured with comparable, although preliminary and limited, results to those of cytological examination. CONCLUSIONS: The preliminary results show that the technique may be feasible for the detection of bladder cancer cells in urine with possible advantages of simplicity, reliability and objectivity. PMID- 17397494 TI - Referral compliance, outcome and predictors of CIN after repeated borderline cervical smears in the Netherlands. AB - BACKGROUND: Borderline cytological abnormalities are diagnosed very frequently but have limited predictive value for high-grade cervical lesions, resulting in high costs, patient anxiety and over treatment. A conservative management strategy for the Dutch diagnostic equivalent of borderline nuclear changes (BNC) was introduced in the Netherlands in 1996, with repeat cytology at 6 and 18 months and referral for colposcopy if BNC is persistent. OBJECTIVE: To analyse compliance with the current guidelines for referral, as well as the outcome after repeated BNC. Concurrently we investigated whether other variables are predictive of high-grade lesions. METHODS: We retrieved 1898 eligible cases of repeated BNC with 4 years follow-up from the national pathology database (PALGA) and performed a nationwide survey. RESULTS: The management strategy for women with repeated BNC in the Netherlands has been accepted and supported. Seventy-seven per cent (77%) of the patients had visited a gynaecologist within 1 year and only 4.3% were lost to follow-up. We found that 25.2% of the patients had a low-grade lesion or worse (CIN 1+) and 10.2% had a high-grade lesion or worse (CIN 2+), among which were four malignancies. The only variable associated with CIN or worse was age. Women under 40 years were found to be at a higher risk. CONCLUSION: This finding may be used for prioritizing women for colposcopy on the basis of their age. More stringent use of the diagnosis of BNC, higher thresholds for colposcopically directed biopsy and introduction of HPV triage, combined with more specific new techniques or combination of techniques such as molecular markers for P16, MIB-1 and L1 may reduce the unnecessary high referral rate and over treatment of healthy women. PMID- 17397495 TI - Inter- and intraobserver reproducibility of thyroid fine needle aspiration cytology: an analysis of discrepant cases. AB - OBJECTIVE: To determine the inter- and intraobserver reproducibility and analyse the discrepant cases of fine needle aspiration cytology (FNAC) of the thyroid. METHODS: Cases of thyroid FNAC with a corresponding histological diagnosis were reviewed regarding the original cytological diagnoses by two observers. The final cytological diagnoses (FCD) included both concordant and consensus diagnoses. The inter- and intraobserver reproducibility and efficacy of thyroid FNAC were calculated based on the FCD. RESULTS: A total of 97 FNAC cases with corresponding histopathological specimens were analysed. Although inter- and intraobserver disagreement in the cytological diagnoses occurred in about one-quarter of the cases analysed (24.7% and 23.7%, respectively), a substantial level of diagnostic interobserver (kappa = 0.71) and intraobserver (kappa = 0.66) reproducibility was observed. The efficacy of the method was 94.4%. Disagreement in the diagnosis was detected in 24 cases (24.7%), most of them (41.7%) for follicular lesions. Discordant cytological diagnoses between the two observers were represented by six (16.2%) of the 37 cases with an FCD of colloid nodule, five (41.7%) of the 12 cases of cellular follicular lesion, all three cases of follicular neoplasm, in two (6.3%) of the 32 cases of PTC, one (16.7%) of six cases of follicular neoplasm with a predominance of Hurthle cells and in one case of poorly differentiated neoplasia. Similarly, major disagreement in intraobserver cytological diagnoses was observed for the diagnosis of follicular lesions: 18 (78.3%) of a total of 23 discordant cases. CONCLUSION: As discrepancies in the cytopathological diagnosis can have repercussions in the management of patients, all cases with a cytological diagnosis of follicular lesions/neoplams should be reviewed in multidisciplinary meetings thus minimizing interobserver variability. PMID- 17397496 TI - Galectin-3 and CD44v6 positivity by RT-PCR method in fine needle aspirates of benign thyroid lesions. AB - OBJECTIVE: To investigate whether the presence of macrophages and Hurthle cells (HC) in benign thyroid lesions could explain the false-positive expression of galectin-3 and CD44v6 detected by reverse transcriptase-polymerase chain reaction (RT-PCR). METHODS: For galectin-3 and CD44v6, RT-PCR was performed on RNA isolated from aspirates obtained by ultrasound guided fine needle aspiration cytology (FNAC) from 123 patients with benign thyroid lesions. The results of RT PCR analysis were evaluated against the definitive FNAC diagnosis. RESULTS: Galectin-3 expression was found in 29% follicular adenoma (FA), 26% Hashimoto thyroiditis (HT), and in 24% nodular goitre (NG). We found a statistically significant relationship between the presence of macrophages and galectin-3 positivity in NG and HT samples (P < 0.0001 and P = 0.0087 respectively). We found a statistically significant (P = 0.0219) relationship between the presence of HC and galectin-3 positivity in HT and a tendency of such a relationship (P = 0.0838) in NG. CD44v6 expression was found in 29% FA, 33% HT and in 18% NG. We found a statistically significant relationship between the presence of HC and positive expression of CD44v6 in NG (P = 0.0003) and a strong tendency of such a relationship in HT (P = 0.0571). We did not find a statistically significant relationship between the presence of macrophages and CD44v6 positivity. In FA, we did not find a statistically significant relationship between the presence of macrophages or HC and galectin-3 or CD44v6 positivity. CONCLUSION: Our results suggest that the presence of macrophages and/or HC may explain the positive expression of galectin-3 and CD44v6 detected by RT-PCR in HT and NG cytological samples. PMID- 17397497 TI - Immunoflow cytometry compared with PCR for the identification of clonality in FNAs of T-cell-rich B-cell lymphomas. AB - OBJECTIVE: The study aimed to compare the utility of immunoflow cytometry (IFC) with two IgH polymerase chain reaction (PCR) methods for the identification of clonality in fine needle aspirations (FNAs) from T-cell-rich B-cell lymphomas (TCRBCLs). METHODS: Ten cases of TCRBCLs were identified in which IFC had been performed according to our previously described method. Seven of these were cases in which the original diagnosis had been made by FNA cytology with IFC and cell block immunohistochemistry (IHC). The remaining three cases only had biopsies with histology, IFC and IHC. Formalin-fixed paraffin-embedded FNA cell block or histology tissue from these specimens had also been submitted for IgH PCR clonality studies using primers FR2a/VLJH and primers FR3a/VLJH. The results were reviewed and compared. RESULTS: All 10 case demonstrated B-cell clonality for at least one of the primer sets on PCR, but none showed light chain restriction on IFC. All TCRBCLs were positive for CD20 and CD79a but negative for CD10 and BCl 2. They were also consistently negative for CD22, CD23, CD5, CD43, ALK-1, cyclin D1 and CD30. CONCLUSIONS: If IgH PCR clonality assays had a turnaround time of 1 or 2 days, there might be a strong case for these studies supporting or even replacing IFC, in the FNA diagnosis of lymphoid lesions. PMID- 17397498 TI - Are all thyroid papillae malignant? An unusual pitfall in FNAC. PMID- 17397499 TI - Spindle cell lipoma: an uncommon tumour with distinctive morphology. PMID- 17397500 TI - Diagnosis of three cases of paracoccidioidomycosis by fine needle aspiration cytology. PMID- 17397501 TI - Reply to Mehrotra et al: ultrasound guided thyroid cytology. PMID- 17397503 TI - Use of a solid absorbent and an accelerant detection canine for the detection of ignitable liquids burned in a structure fire. AB - Ignitable Liquid Absorbent (ILA), a commercial solid absorbent intended to assist fire scene investigators in sample location and collection, has been field tested in three separate room fires. The ability of the ILA to detect and absorb different amounts of gasoline, odorless paint thinner, and camp fuel on two different substrates after a full-scale burn was assessed against results from an accelerant detection canine and laboratory analysis using gas chromatography-mass spectrometry (GC-MS). The canine correctly alerted on most of the panels that contained an ignitable liquid after the fire, while the ILA indicator dye failed to indicate in the presence of gasoline and camp fuel. GC-MS results for ignitable liquid residue from each panel and from the ILA showed that ILA absorbed odorless paint thinner and camp fuel from most of the test panels, but failed to absorb gasoline from the panels on which gasoline was confirmed to be present. PMID- 17397504 TI - Taphonomic changes to blunt force trauma: a preliminary study. AB - This study examines the effects of taphonomic processes on blunt force trauma (BFT) through an experimental study involving pig heads. Of particular concern is the possibility that taphonomic changes can create pseudo-trauma and/or conceal evidence of actual trauma. BFT was inflicted on 10 pig skulls using a hammer. The skulls were subsequently exposed to the environment for 12 months. Seven taphonomic changes were evaluated: the freeze-thaw cycle; rodent gnawing; carnivore scavenging; presence/weight of soil; presence/weight of rain and snow; movement/displacement of bones; and discoloration due to sun bleaching and grass staining. Taphonomic effects varied between cancellous, compact, fresh, and degreased bone. Freezing and thawing, exposure to rain and snow, movement of the skulls, and soil erosion altered and, in some cases disguised, pre-existing trauma. Rodent and carnivore activity did not obliterate evidence of BFT. Recommendations for evaluating BFT on remains affected by taphonomic processes are presented. As each taphonomic process outlined by this study has the potential to disguise antemortem injury, the authors propose that one must carefully examine large, circular openings in the skull that may represent the remnant evidence of BFT. PMID- 17397505 TI - The uniqueness of the human anterior dentition: a geometric morphometric analysis. AB - The analysis of bitemarks has a significant bearing on forensic odontology and has attracted an increasingly sophisticated array of techniques in its evaluation. Two postulates underlie all bitemark analyses: firstly, that the characteristics of the anterior teeth involved in the bite are unique, and secondly, that this uniqueness is accurately recorded in the material bitten. Here, we investigate the question of the uniqueness of the anterior dentition. To do this, we use geometric morphometric techniques based on landmark and semilandmark data. The incisor and canine occlusal surfaces of 50 randomly selected orthodontic casts of young individuals (17-20 years) of both sexes form the material for this study. We analyzed the sizes of these teeth by means of landmark and semilandmark analysis to calculate Procrustes distances between tooth outlines. In order to analyze shape variation among individuals, we carried out principal components analyses on the partial warp scores. These are derived from Partial Procrustes coordinates aligned by means of thin-plate spline decomposition based on the bending energy matrix. The results indicate that there is no sexual dimorphism in the shape of the upper or lower arches. Plots of centroid size and first relative warps show less superposition among individuals than in shape analysis. This means that, when the size and shape are considered together, the difference between arches increases. Procrustes superimposition between the two individuals located most closely (0.0444) and the two most separated (0.1567) along the first axis of relative warp analyses show that individuals are not only differentiated by the relative position of their teeth but also by their arch shape. In conclusion, it appears that the incisal surfaces of the anterior dentition are in fact unique. PMID- 17397506 TI - A leucine-rich repeat protein is required for growth promotion and enhanced seed production mediated by the endophytic fungus Piriformospora indica in Arabidopsis thaliana. AB - Piriformospora indica, a basidiomycete of the Sebacinaceae family, promotes the growth, development and seed production of a variety of plant species. Arabidopsis plants colonized with the fungus produce 22% more seeds than uncolonized plants. Deactivating the Arabidopsis single-copy gene DMI-1, which encodes an ion carrier required for mycorrihiza formation in legumes, does not affect the beneficial interaction between the two symbiotic partners. We used cellular and molecular responses initiated during the establishment of the interaction between P. indica and Arabidopsis roots to isolate mutants that fail to respond to the fungus. An ethylmethane sulfonate mutant (Piriformospora indica insensitive-2; pii-2), and a corresponding insertion line, are impaired in a leucine-rich repeat protein (At1g13230). The protein pii-2, which contains a putative endoplasmic reticulum retention signal, is also found in Triton X-100 insoluble plasma membrane microdomains, suggesting that it is present in the endoplasmic reticulum/plasma membrane continuum in Arabidopsis roots. The microdomains also contain an atypical receptor protein (At5g16590) containing leucine-rich repeats, the message of which is transiently upregulated in Arabidopsis roots in response to P. indica. This response is not detectable in At1g13230 mutants, and the protein is not detectable in the At1g13230 mutant microdomains. Partial deactivation of a gene for a sphingosine kinase, which is required for the biosynthesis of sphingolipid found in plasma membrane microdomains, also affects the Arabidopsis/P. indica interaction. Thus, pii-2, and presumably also At5g16590, two proteins present in plasma membrane microdomains, appear to be involved in P. indica-induced growth promotion and enhanced seed production in Arabidopsis thaliana. PMID- 17397507 TI - Increase in CPD photolyase activity functions effectively to prevent growth inhibition caused by UVB radiation. AB - Rice cultivars vary widely in their sensitivity to ultraviolet B (UVB) and this has been correlated with cyclobutane pyrimidine dimer (CPD) photolyase mutations that alter the structure/function of this photorepair enzyme. Here, we tested whether CPD photolyase function determines the UVB sensitivity of rice (Oryza sativa) by generating transgenic rice plants bearing the CPD photolyase gene of the UV-resistant rice cultivar Sasanishiki in the sense orientation (S-B and S-C lines) or the antisense orientation (AS-D line). The S-B and S-C plants had 5.1- and 45.7-fold higher CPD photolyase activities than the wild-type, respectively, were significantly more resistant to UVB-induced growth damage, and maintained significantly lower CPD levels in their leaves during growth under elevated UVB radiation. Conversely, the AS-D plant had little photolyase activity, was severely damaged by elevated UVB radiation, and maintained higher CPD levels in its leaves during growth under UVB radiation. Notably, the S-C plant was not more resistant to UVB-induced growth inhibition than the S-B plant, even though it had much higher CPD photolyase activity. These results strongly indicate that UVB induced CPDs are one of principal causes of UVB-induced growth inhibition in rice plants grown under supplementary UVB radiation, and that increasing CPD photolyase activity can significantly alleviate UVB-caused growth inhibition in rice. However, further protection from UVB-induced damage may require the genetic enhancement of other systems as well. PMID- 17397508 TI - SA-inducible Arabidopsis glutaredoxin interacts with TGA factors and suppresses JA-responsive PDF1.2 transcription. AB - Salicylic acid (SA) is a plant signaling molecule that mediates the induction of defense responses upon attack by a variety of pathogens. Moreover, it antagonizes gene induction by the stress signaling molecule jasmonic acid (JA). Several SA responsive genes are regulated by basic/leucine zipper-type transcription factors of the TGA family. TGA factors interact with NPR1, a central regulator of many SA induced defense responses including SA/JA antagonism. In order to identify further regulatory proteins of SA-dependent signaling pathways, a yeast protein interaction screen with tobacco TGA2.2 as bait and an Arabidopsis thaliana cDNA prey library was performed and led to the identification of a member of the glutaredoxin family (GRX480, encoded by At1g28480). Glutaredoxins are candidates for mediating redox regulation of proteins because of their capacity to catalyze disulfide transitions. This agrees with previous findings that the redox state of both TGA1 and NPR1 changes under inducing conditions. Transgenic Arabidopsis plants ectopically expressing GRX480 show near wild-type expression of standard marker genes for SA- and xenobiotic-inducible responses. In contrast, transcription of the JA-dependent defensin gene PDF1.2 was antagonized by transgenic GRX480. This, together with the observation that GRX480 transcription is SA-inducible and requires NPR1, suggests a role of GRX480 in SA/JA cross-talk. Suppression of PDF1.2 by GRX480 depends on the presence of TGA factors, indicating that the GRX480/TGA interaction is effective in planta. PMID- 17397509 TI - SDE5, the putative homologue of a human mRNA export factor, is required for transgene silencing and accumulation of trans-acting endogenous siRNA. AB - Post-transcriptional gene silencing (PTGS) is a sequence-specific RNA degradation process conserved in fungi, plants and animals. The trigger of the mechanism is double-stranded RNA derived from transgenic or endogenous loci and formed by intra- or inter-molecular interactions of single-stranded RNAs or the action of RNA-dependent RNA polymerases (RDRs). Double-stranded RNA from various sources is processed by one of the four Dicer-like (DCL) proteins in Arabidopsis, and the resulting short RNAs enter into at least four different pathways, one of which involves the production of trans-acting short interfering RNAs (tasiRNAs). We report here a novel gene (SDE5) that is required for transgene silencing and the production of tasiRNAs. Mutation in SDE5 also results in hyper-susceptibility to cucumber mosaic virus but not turnip mosaic virus. However, like RDR6, SDE5 is not involved in inverted repeat-induced transgene silencing or the biogenesis of microRNAs and 24 nt siRNAs produced by DCL3. Based on these results, we propose that SDE5 acts together with RDR6 in generating double-stranded RNA from specific single-stranded RNAs. As the sequence of SDE5 has sequence features shared by TAP, a human mRNA export factor, we propose that its role could be in the transport of RNA molecules that are converted into a double-stranded form by RDR6. PMID- 17397510 TI - A method for cellular localization of gene expression via quantitative in situ hybridization in plants. AB - A quantitative in situ hybridization technique (quantitative whole-mount in situ hybridization, QISH) for plants is described. It employs direct hybridization of fluorescently labelled gene-specific oligonucleotides in large tissue pieces combined with optical sectioning. It dramatically increases the throughput compared with conventional antibody- and microtome-based in situ mRNA hybridization methods, while simultaneously eliminating artefact-prone preparation steps that prevent reliable quantification in conventional methods. The key feature of this technique is the quantification of gene expression using housekeeping genes (cytosolic GAPDH and 18S RNA) as internal standards. This feature enables a correction of varying cytoplasm/vacuole ratios in different cell types, as well as tissue optical effects and non-specific signals. The quantitative nature of the technique allows for analysis of gene expression in response to different environmental conditions, as well as tissue- and age dependent differences in gene expression patterns. In addition to testing tissue permeabilization, structural preservation, specificity, linearity and tissue optical effects, we verified the reliability of the technique with three Arabidopsis thaliana genes of known function and distribution. These were the rbcL gene for ribulose 1,5-bisphosphate carboxylase, the developmentally related gene SCARECROW (AtSCR) and PHOT-1, a photoreceptor kinase. As expected, rbcL mRNA was found in all photosynthetic cells, while SCR mRNA was detected mainly in bundle sheath cells and PHOT-1 was found predominantly in epidermal and cortical cells of the apical hook of light-grown seedlings. As an application example, QISH was used to measure transcript abundance for a zinc transporter from the ZIP family of transporters in the Zn/Cd hyperaccumulator model plant, Thlaspi caerulescens, and the related non-accumulator Thlaspi arvense. This showed that QISH can be used to compare differences in mRNA levels between cell types, plant growth conditions and plant species. Messenger RNA for the zinc transporter gene ZNT1 was abundant in photosynthetic cells, but not in the epidermal storage cells where metal hyperaccumulation in T. caerulescens occurs. This indicates that ZNT1 does not directly participate in metal hyperaccumulation within the leaf. Growing T. caerulescens with high zinc levels strongly reduced ZNT1 transcript abundance in the spongy mesophyll cells, but less in the other cell types. In T. arvense, ZNT1 mRNA levels were generally much lower, and were furthermore drastically reduced by growth at increased zinc levels, confirming earlier reports regarding ZNT1 regulation in these two Thlaspi species. PMID- 17397511 TI - Protection of estrogens against the progression of chronic liver disease. AB - Hepatitis C virus infections are recognized as a major causative factor of chronic liver disease. A characteristic feature of chronic hepatitis C, alcoholic liver disease and non-alcoholic fatty liver disease is hepatic steatosis. Hepatic steatosis leads to an increase in lipid peroxidation in hepatocytes, which, in turn, activates hepatic stellate cells (HSCs). HSCs are also thought to be the primary target cells for inflammatory and oxidative stimuli, and to produce extracellular matrix components. Based on available clinical information, chronic hepatitis C appears to progress more rapidly in men than in women, and cirrhosis is predominately a disease of men and postmenopausal women. Estradiol is a potent endogenous antioxidant. Hepatic steatosis was reported to become evident in an aromatase-deficient mouse and was diminished in animals after treatment with estradiol. Our previous studies showed that estradiol suppressed hepatic fibrosis in animal models, and attenuated HSC activation by suppressing the generation of reactive oxygen species in primary cultures. Variant estrogen receptors were found to be expressed to a greater extent in male patients with chronic liver disease than in female subjects. A better understanding of the basic mechanisms underlying the gender-associated differences observed in the progression of chronic liver disease would provide valuable information relative to the search for effective antifibrogenic therapies. PMID- 17397512 TI - Interleukin-17 as a new marker of severity of acute hepatic injury. AB - AIM: To determine cytokines associated with the progression of acute hepatic injury (AHI), we comprehensively evaluated the serum levels of 17 cytokines. METHODS: We simultaneously measured serum levels of 17 cytokines on admission using a newly developed suspension array protein assay system in 51 patients with AHI, including 15 conventional AHI (CAHI), 15 severe AHI (SAHI) and 21 fulminant hepatic failure (FHF). RESULTS: Interleukin (IL)-6, IL-8 and IL-17 levels were significantly different among the three disease types as determined by one-way analysis of variance, and only the IL-17 level showed a significant elevation in SAHI and FHF than in CAHI. Namely, the IL-17 levels in SAHI and FHF patients were 4.4 (2.0-11.0) (mean [1 .s.d. range]) and 5.6 (2.0-18.5) pg/mL, respectively, whereas all CAHI patients showed levels lower than the lower limit of detection (2.0 pg/mL). In multiple regression analysis for each factor of model for end stage liver disease (MELD) score, only IL-10 level was selected as the significant independent variable for total bilirubin level, only IL-17 level for prothrombin time, and TNF-alpha and IL-1beta levels for creatinine level. CONCLUSION: These data suggest the usefulness of serum IL-17 level in evaluating the severity of AHI, thus emphasizing the necessity for the basic investigation of the pathological role of IL-17 in acute hepatitis. PMID- 17397513 TI - Characteristics of geographic distributions and route of infection for hepatitis B virus genotype D in Ehime area in western Japan. AB - AIMS: Hepatitis B virus genotype D (HBV/D) is rare in Japan, and has been shown to circulate in Ehime prefecture in western Japan. HBV/D is suspected to have been transferred into Ehime from Russia as a result of the Japanese-Russian War. This study examined the current geographic spread and infectious route for HBV/D in Ehime. METHODS: HBV genotype was determined for 508 patients with chronic HBV infection and 46 patients with acute HBV infection hepatitis (acute hepatitis, AH), all of whom were living in Ehime. Ehime was divided into three areas and genotypic distributions were studied. RESULTS: The ratio of genotypes A,B,C and D in chronically infected patients were 1.8%, 4.5%, 87.8% and 5.9%, respectively. Most patients chronically infected with HBV/D lived in the central area. Only two patients lived in the east and south-west areas, and both had lived in the central area in childhood. Patients with AH due to HBV/D were found only in the central area. CONCLUSION: HBV/D has not yet spread widely to areas other than central Ehime, although small numbers of infected people have moved to other areas. The major infectious route for patients with AH is sexual transmission, regardless of HBV genotype. PMID- 17397514 TI - Correlation of hepatic steatosis with body mass index, serum ferritin level and hepatic fibrosis in Japanese patients with chronic hepatitis C. AB - AIMS: The present study was aimed at determining the predictors of hepatic steatosis and fibrosis in Japanese patients with chronic hepatitis C. METHODS: The relationship between the degrees of hepatic steatosis or fibrosis and several clinical parameters was evaluated using univariate and multivariate analyses. RESULTS: Steatosis was observed in 117 out of 184 patients (64%), including 45 patients (25%) with grade 1 (<10% of hepatocytes affected), 56 patients (30%) with grade 2 (10-30%), 12 patients (7%) with grade 3 (30-50%), and four patients (2%) with grade 4 (>50%). In the multivariate analysis, body mass index (BMI) (P = 0.0038) and serum ferritin (P < 0.0001) were selected as independent predictors of hepatic steatosis. Six of the 184 patients (3%) had stage 0 fibrosis (no fibrosis), 87 patients (47%) had stage 1, 55 patients (30%) had stage 2 and 36 patients (20%) had stage 3. In the multivariate analysis, platelet count (P = 0.0012), aspartate aminotransferase (AST) (P = 0.0219), hyaluronic acid (P < 0.0001) and the grade of steatosis (P = 0.0008) were selected as independent predictors of hepatic fibrosis. CONCLUSION: Obesity and iron storage, as evaluated by BMI and serum ferritin level, respectively, have important roles in the pathogenesis of hepatic steatosis, which is a factor responsible for the development of hepatic fibrosis in Japanese patients with chronic hepatitis C. PMID- 17397515 TI - Mapping of a disease susceptibility locus in the HLA region for Primary Biliary Cirrhosis in Japan. AB - AIMS: Primary biliary cirrhosis (PBC) is a chronic progressive cholestatic disease of unknown pathogenesis. However, several reports have demonstrated the involvement of genetic backgrounds in this syndrome. The aim of this study is to examine the genetic disequilibrium in the HLA region in Japanese patients using six microsatellite markers. METHODS: Genomic DNAs were obtained from 73 patients with PBC (patient cohort) and 186 healthy volunteers (control cohort). Genetic polymorphisms at six microsatellite markers (D6S1568, DQ.CAR, D6S273, TNF-d, C1-2 A, C3-2-11) were determined using fluorescence-labeled polymerase chain reaction (PCR) genetic analyzer. Allele frequencies were estimated by direct counting and the genotypic differentiation test was performed by the Markov chain method using Genepop software. RESULTS: Among these six microsatellite markers, four markers in the patients significantly (P < 0.05) deviated from the Hardy-Weinberg equilibrium: DQ.CAR (P = 0.0278), D6S273 (P = 0.0168), TNF-d (P = 0.0089) and C1 2-A (P = 0.0005). Genotypic differentiation test between the patients and controls demonstrated that DQ.CAR (P = 0.0111), TNF-d (P = 0.0051) and C1-2-A (P = 0.0371) were significant. Finally, allelic association test revealed before correction for multiple testing demonstrated allele125 of TNF-d (P = 0.00065, Pc = 0.0052) and allele246 of C1-2-A (P = 0.0026 Pc = 0.033) had significant association after Bonferroni's correction. CONCLUSION: Disequilibrium mapping using microsatellite markers was a useful method to narrow a disease susceptibility locus. The possible susceptibility gene in the HLA region is thought to be localized around or in the TNF gene. Further studies seem feasible using more closely distributed microsatellite markers or single nucleotide polymorphisms (SNPs) to narrow the susceptibility locus in PBC in Japanese populations. PMID- 17397516 TI - Impaired cytotoxic T lymphocyte inductivity by dendritic cells derived from patients with hepatitis C virus-positive hepatocellular carcinoma. AB - AIM: Peptide-based therapeutic vaccines are being developed. The aim of this study was to determine the feasibility of immunotherapy to hepatitis C virus (HCV)-positive hepatocellular carcinoma (HCC) by assessing the inductivity of peptide-specific cytotoxic T lymphocyte (CTL) by dendritic cells. METHODS: The inductivity of CTL was characterized in six patients with HCV-positive HCC, and compared to seven healthy volunteers and six patients with chronic HCV hepatitis (control). RESULTS: Peptide-specific CTL was comparably induced in controls, but not induced in any patients with HCC. To characterize this, the cytokine profile and the expression of surface molecules interacting between dendritic and T cells were evaluated. Among the cytokines, production of interferon (IFN)-gamma was found to be impaired and closely related to the results of CTL assays, while the expression of surface molecules showed no significant changes. CONCLUSIONS: In HCV-positive HCC patients, CTL inductivity by dendritic cells is impaired. This may be related to the impaired production of IFN-gamma. PMID- 17397517 TI - Amniotic epithelial cell-derived cholangiocytes in experimental cholestatic ductal hyperplasia. AB - AIM: Bile duct paucity, ductopenia, is a feature of end-stage chronic cholangiopathies such as primary biliary cirrhosis. The limited proliferative ability of cholangiocytes after specific injury is thought to be the principal cause of ductopenia, although the detailed mechanisms involved are unclear. It has been reported that human amniotic epithelial cells (AEC) express differentiation markers of hepatic parenchymal cells, suggesting a resemblance of AEC to hepatic progenitor cells. The aim of the present study was to develop a mouse model of experimental cholestasis to assess the capability of mouse AEC to trans-differentiate into cholangiocytes. METHODS: Enhanced green fluorescent protein (EGFP)-transgenic C57BL/6 pregnant female mice were used as the source of AEC. At 11.5 gestational days, 1 x 10(5) AEC were isolated from EGFP-transgenic mouse embryos and transferred into C57BL/6 mice. Chronic cholestasis was induced by 0.1%alpha-naphthylisothiocyanate (ANIT) feeding immediately after the transfer of AEC. The proliferation of cholangiocytes in the livers was assessed morphologically and immunohistochemically (cytokeratin 7; CK7). The proliferative activity was also quantified immunohistochemically by proliferating cell nuclear antigen (PCNA) protein expression. EGFP of transferred AEC was confirmed by fluorescent laser microscopy and immunofluorescent staining for EGFP. Also, Notch2 and Hes1 expression was evaluated to examine the roles of the differentiation markers in this process. RESULTS: Marked proliferation of cholangiocytes was observed in ANIT-fed mice confirmed by quantitative CK7 (3-4 fold vs control) and PCNA (11-20 fold vs control) staining. EGFP and CK7 double positive cells in interlobular bile ducts were confirmed in the livers of AEC transferred recipients. Positivity of EGFP was further confirmed by the immunofluorescent staining for EGFP. Moreover, both Notch2 and Hes1 expression was confirmed in the proliferative bile duct in this model. CONCLUSIONS: Significant ductular proliferation was observed in ANIT-fed mice. EGFP-positive cholangiocytes were confirmed in this chronic cholestasis model. AEC transfer was able to contribute to the repopulating of proliferating cholangiocytes under cholestasis, suggesting AEC might be a candidate cell source for stem cell administration in future clinical applications to re-model interlobular bile ducts. PMID- 17397518 TI - Increased TFF1 trefoil protein expression in Opisthorchis viverrini-associated cholangiocarcinoma is important for invasive promotion. AB - AIMS: Cholangiocarcinoma (CCA) is a poor prognosis cancer that presents with metastatic disease. This cancer expresses MUC5AC, a mucin which normally co expresses with trefoil factor family 1 (TFF1) protein. TFF1 is a signalling protein that can activate epithelial cell invasion and has been considered as a metastasis stimulating agent. The aim of this study was to determine the co expression of TFF1 and MUC5AC in CCA tissues and examine the activity of TFF1 for stimulating the invasive property of CCA cell lines. METHODS: In this study, TFF1 and MUC5AC were detected in CCA tissues by using immunohistochemistry. The correlations of both proteins expression with clinical data were analyzed. The activity of TFF1 was investigated using an in vitro invasion assay with established CCA cell lines KKU-100 and KKU-M213. RESULTS: We demonstrated a high level of expression of TFF1 in 91.80% of CCA that is associated with a high level of co-expression with MUC5AC in 80.33% of cases. In vitro invasion assay showed that both cell lines have similar responses to TFF1 that could act as both a chemokinetic and chemotactic agent. The dose-response curves were bell-shaped. CONCLUSION: TFF1 showed co-expression with MUC5AC in CCA tissues and invasive stimulating activity in vitro. These results may indicate a role for TFF1 in promoting tumor invasion in CCA. PMID- 17397519 TI - Preliminary experience for reducing biliary complication in adult-to-adult living donor liver transplantation using right lobe graft. AB - AIM: To report the author's preliminary experience with adult-to-adult living donor liver transplantation (LDLT) using right lobe liver grafts. METHODS: From January 2002 to July 2006, 50 adult patients underwent LDLT using right lobe grafts at West China Hospital, Sichuan University Medical School, China. At present, 24 recipients have been followed up for over nine months. RESULTS: There were no deaths amongst the donors. Biliary reconstruction for 35 hepatic duct orifices from 24 donor grafts was performed, which included 19 reconstructions of duct-to-duct anastomoses and 16 cholangio-jejunostomy. Two recipients had biliary complications including one bile leakage and one slight biliary stricture. CONCLUSION: With the improved methods for biliary reconstruction, we achieved good results in 24 recipients of adult-to-adult LDLT. We consider that microsurgical technique and fixed operator could decrease the biliary complications of LDLT. PMID- 17397520 TI - Expression genomics in breast cancer research: microarrays at the crossroads of biology and medicine. AB - Genome-wide expression microarray studies have revealed that the biological and clinical heterogeneity of breast cancer can be partly explained by information embedded within a complex but ordered transcriptional architecture. Comprising this architecture are gene expression networks, or signatures, reflecting biochemical and behavioral properties of tumors that might be harnessed to improve disease subtyping, patient prognosis and prediction of therapeutic response. Emerging 'hypothesis-driven' strategies that incorporate knowledge of pathways and other biological phenomena in the signature discovery process are linking prognosis and therapy prediction with transcriptional readouts of tumorigenic mechanisms that better inform therapeutic options. PMID- 17397521 TI - A day in the life of a genome biologist in the not-too-distant future. AB - A look into the future of a biologist: daily activities governed by presidential mandates and acts. PMID- 17397522 TI - Hox go omics: insights from Drosophila into Hox gene targets. AB - Genetic studies of the targets of the Hox genes have revealed only the tip of the iceberg. Recent microarray studies that have identified hundreds more transcriptional responses to Hox genes in Drosophila will help elucidate the role of Hox genes in development and evolution. PMID- 17397523 TI - Is an aggressive approach justified in the management of an aggressive cancer- the squamous cell carcinoma of thyroid? AB - BACKGROUND: Primary squamous cell carcinoma of the thyroid is an extremely rare neoplasm, with less than 50 cases reported in the world literature. The prognosis is poor with a median survival of less than six months. The death is usually secondary to progression of local disease as distant metastases are rare. CASE REPORTS: Three cases, two males and one female presenting with sudden increase in the size of long standing thyroid swellings and associated pressure effects on the aero-digestive tract are reported. Exhaustive clinical, endoscopic, and radiological examinations did not reveal any primary site of squamous-cell carcinoma as the likely source of the metastases, or of any contiguous spread from neighboring structures. Two cases were managed by combined modality therapy including curative surgery with radiotherapy and one by radiotherapy alone. CONCLUSION: Primary squamous cell carcinoma is a rare malignancy with a poor outcome inspite of combined modality therapy. Out of three reported cases, two succumbed to their disease within less than one year. Aggressive surgery in the form of curative resection along with adjuvant radiotherapy is recommended, the tumor being chemo resistant. PMID- 17397524 TI - Qualitative and quantitative dermatoglyphic traits in patients with breast cancer: a prospective clinical study. AB - BACKGROUND: Breast cancer is one of the most extensively studied cancers and its genetic basis is well established. Dermatoglyphic traits are formed under genetic control early in development but may be affected by environmental factors during first trimester of pregnancy. They however do not change significantly thereafter, thus maintaining stability not greatly affected by age. These patterns may represent the genetic make up of an individual and therefore his/her predisposition to certain diseases. Patterns of dermatoglyphics have been studied in various congenital disorders like Down's syndrome and Kleinfelter syndrome. The prints can thus represent a non-invasive anatomical marker of breast cancer risk and thus facilitate early detection and treatment. METHODS: The study was conducted on 60 histo-pathologically confirmed breast cancer patients and their digital dermatoglyphic patterns were studied to assess their association with the type and onset of breast cancer. Simultaneously 60 age-matched controls were also selected that had no self or familial history of a diagnosed breast cancer and the observations were recorded. The differences of qualitative (dermatoglyphic patterns) data were tested for their significance using the chi-square test, and for quantitative (ridge counts and pattern intensity index) data using the t- test. RESULTS: It was observed that six or more whorls in the finger print pattern were statistically significant among the cancer patients as compared to controls. It was also seen that whorls in the right ring finger and right little finger were found increased among the cases as compared to controls. The differences between mean pattern intensity index of cases and controls were found to be statistically significant. CONCLUSION: The dermatoglyphic patterns may be utilized effectively to study the genetic basis of breast cancer and may also serve as a screening tool in the high-risk population. In a developing country like India it might prove to be an anatomical, non-invasive, inexpensive and effective tool for screening and studying the patterns in the high-risk population. PMID- 17397525 TI - Protocol for the PACE trial: a randomised controlled trial of adaptive pacing, cognitive behaviour therapy, and graded exercise, as supplements to standardised specialist medical care versus standardised specialist medical care alone for patients with the chronic fatigue syndrome/myalgic encephalomyelitis or encephalopathy. AB - BACKGROUND: Chronic fatigue syndrome (CFS, also called myalgic encephalomyelitis /encephalopathy or ME) is a debilitating condition with no known cause or cure. Improvement may occur with medical care and additional therapies of pacing, cognitive behavioural therapy and graded exercise therapy. The latter two therapies have been found to be efficacious in small trials, but patient organisations surveys have reported adverse effects. Although pacing has been advocated by patient organisations, it lacks empirical support. Specialist medical care is commonly provided but its efficacy when given alone is not established. This trial compares the efficacy of the additional therapies when added to specialist medical care against specialist medical care alone. METHODS: 600 patients, who meet operationalised diagnostic criteria for CFS, will be recruited from secondary care into a randomised trial of four treatments, stratified by current co morbid depressive episode and different CFS/ME criteria. The four treatments are standardised specialist medical care either given alone, or with adaptive pacing therapy or cognitive behaviour therapy or graded exercise therapy. Supplementary therapies will involve fourteen sessions over 23 weeks and a booster session at 36 weeks. Outcome will be assessed at 12, 24, and 52 weeks after randomisation. Two primary outcomes of self-rated fatigue and physical function will assess differential effects of each treatment on these measures. Secondary outcomes include adverse events and reactions, subjective measures of symptoms, mood, sleep and function and objective measures of physical activity, fitness, cost-effectiveness and cost-utility. The primary analysis will be based on intention to treat and will use logistic regression models to compare treatments. Secondary outcomes will be analysed by repeated measures analysis of variance with a linear mixed model. All analyses will allow for stratification factors. Mediators and moderators will be explored using multiple linear and logistic regression techniques with interactive terms, with the sample split into two to allow validation of the initial models. Economic analyses will incorporate sensitivity measures. DISCUSSION: The results of the trial will provide information about the benefits and adverse effects of these treatments, their cost-effectiveness and cost-utility, the process of clinical improvement and the predictors of efficacy. PMID- 17397526 TI - The New Zealand Asthma and Allergy Cohort Study (NZA2CS): assembly, demographics and investigations. AB - BACKGROUND: Asthma and allergy are highly prevalent in industrialised countries. Longitudinal and cross-sectional studies have identified a number of potential risk factors for these conditions, including genetic and environmental factors, with significant gene-environment relationships. Birth cohort studies have been proposed as an important tool to explore these risk factors, particularly exposures in early life that are associated with later disease or protection from disease. This paper describes the establishment of a birth cohort in New Zealand. METHODS: A birth cohort was established in 1996 in Christchurch and Wellington and infants recruited between 1997-2001. Expectant mothers were recruited by midwives. Children and mothers have undergone assessment by serial questionnaires, environmental assessment including mould and allergen exposure, skin-prick testing, and at age six years are undergoing full assessment for the presence of asthma, atopy and allergic disease, including genetic assessment. RESULTS: A total of 1105 children have been recruited, and the retention rate at fifteen months was 91.4%. 15.2% of the children at recruitment have been identified as Maori. A positive family history of asthma, eczema or hay fever has been reported in 84% of children. All children have now been assessed at fifteen months and 685 children from the cohort have reached age six years and have completed the six year assessment. CONCLUSIONS: The cohort is fully assembled, and assessment of children is well advanced, with good retention rates. The study is well placed to address many current hypotheses about the risk factors for allergic disease and asthma. PMID- 17397527 TI - Wild food plants of popular use in Sicily. AB - In the present work the authors report the result of their food ethnobotanical researches, which have been carried out in Sicily during the last thirty years. Data concerning 188 wild species used in the traditional Sicilian cuisine are reported. The authors underline those species that are partially or completely unknown for their culinary use and they illustrate other species that local inhabitants suggested in the prevention or treatment of symptomatologies caused by a refined diet, poor in vegetables. These data want to contribute to avoid the loss of traditional knowledge on uses and recipes concerning wild food botanicals, and to encourage further studies for those species that have not yet been sufficiently researched in their food chemical and nutritional profile. These studies may also suggest new applications for a few botanicals in medico nutritional fields. The work includes also a short review of the seaweeds and mushrooms traditionally gathered and consumed in Sicily. PMID- 17397528 TI - FGFR1 amplification in breast carcinomas: a chromogenic in situ hybridisation analysis. AB - BACKGROUND: The amplicon on 8p11.2 is reported to be found in up to 10% of breast carcinomas. It has been demonstrated recently that this amplicon has four separate cores. The second core encompasses important oncogene candidates, including the fibroblast growth factor receptor 1 (FGFR1) gene. Recent studies have demonstrated that specific FGFR1 amplification correlates with gene expression and that FGFR1 activity is required for the survival of a FGFR1 amplified breast cancer cell line. METHODS: FGFR1 amplification was analysed in tissue microarrays comprising a cohort of 880 unselected breast tumours by means of chromogenic in situ hybridisation using inhouse-generated FGFR1-specific probes. Chromogenic in situ hybridisation signals were counted in a minimum 30 morphologically unequivocal neoplastic cells. Amplification was defined as >5 signals per nucleus in more than 50% of cancer cells or when large gene copy clusters were seen. RESULTS: FGFR1 amplification was observed in 8.7% of the tumours and was significantly more prevalent in patients >50 years of age and in tumours that lacked HER2 expression. No association was found with other histological parameters. Survival analysis revealed FGFR1 amplification as an independent prognostic factor for overall survival in the whole cohort. Subgroup analysis demonstrated that the independent prognostic impact of FGFR1 amplification was only seen in patients with oestrogen-receptor-positive tumours, where FGFR1 amplification was the strongest independent predictor of poor outcome. CONCLUSION: Given that up to 8.7% of all breast cancers harbour FGFR1 amplification and that this amplification is an independent predictor of overall survival, further studies analysing the FGFR1 as a potential therapeutic target for breast cancer patients are warranted. PMID- 17397529 TI - Inborn errors in the metabolism of glutathione. AB - Glutathione is a tripeptide composed of glutamate, cysteine and glycine. Glutathione is present in millimolar concentrations in most mammalian cells and it is involved in several fundamental biological functions, including free radical scavenging, detoxification of xenobiotics and carcinogens, redox reactions, biosynthesis of DNA, proteins and leukotrienes, as well as neurotransmission/neuromodulation. Glutathione is metabolised via the gamma glutamyl cycle, which is catalyzed by six enzymes. In man, hereditary deficiencies have been found in five of the six enzymes. Glutathione synthetase deficiency is the most frequently recognized disorder and, in its severe form, it is associated with hemolytic anemia, metabolic acidosis, 5-oxoprolinuria, central nervous system (CNS) damage and recurrent bacterial infections. Gamma glutamylcysteine synthetase deficiency is also associated with hemolytic anemia, and some patients with this disorder show defects of neuromuscular function and generalized aminoaciduria. Gamma-glutamyl transpeptidase deficiency has been found in patients with CNS involvement and glutathionuria. 5-Oxoprolinase deficiency is associated with 5-oxoprolinuria but without a clear association with other symptoms. Dipeptidase deficiency has been described in one patient. All disorders are very rare and inherited in an autosomal recessive manner. Most of the mutations are leaky so that many patients have residual enzyme activity. Diagnosis is made by measuring the concentration of different metabolites in the gamma-glutamyl cycle, enzyme activity and in glutathione synthetase and gamma glutamylcysteine synthetase deficiency, also by mutation analysis. Prenatal diagnosis has been preformed in glutathione synthetase deficiency. The prognosis is difficult to predict, as few patients are known, but seems to vary significantly between different patients. The aims of the treatment of glutathione synthesis defects are to avoid hemolytic crises and to increase the defense against reactive oxygen species. No treatment has been recommended for gamma-glutamyl transpeptidase, 5-oxoprolinase and dipeptidase deficiency. PMID- 17397530 TI - Evaluation of gene-expression clustering via mutual information distance measure. AB - BACKGROUND: The definition of a distance measure plays a key role in the evaluation of different clustering solutions of gene expression profiles. In this empirical study we compare different clustering solutions when using the Mutual Information (MI) measure versus the use of the well known Euclidean distance and Pearson correlation coefficient. RESULTS: Relying on several public gene expression datasets, we evaluate the homogeneity and separation scores of different clustering solutions. It was found that the use of the MI measure yields a more significant differentiation among erroneous clustering solutions. The proposed measure was also used to analyze the performance of several known clustering algorithms. A comparative study of these algorithms reveals that their "best solutions" are ranked almost oppositely when using different distance measures, despite the found correspondence between these measures when analysing the averaged scores of groups of solutions. CONCLUSION: In view of the results, further attention should be paid to the selection of a proper distance measure for analyzing the clustering of gene expression data. PMID- 17397531 TI - A focus reduction neutralization assay for hepatitis C virus neutralizing antibodies. AB - BACKGROUND/AIM: The role of humoral immunity in hepatitis C virus (HCV) infection is poorly understood. Nevertheless, there is increasing interest in characterizing the neutralizing antibodies in the serum of HCV-infected patients. Focus reduction assays have been widely used to evaluate neutralizing antibody responses against a range of non-cytopathic viruses. Based on the recent development of a HCV cell culture system using the genotype 2 JFH-1-strain, we developed a focus reduction assay for HCV-neutralizing antibodies. METHODS: The focus reduction assay was based on a standard microneutralization assay in which immunostained foci on tissue culture plates are counted. The neutralizing anti HCV antibodies titers of purified serum immunoglobulin samples from seventy-seven individuals were determined using a 50% focus reduction neutralization assay. Each titer was determined as the log value of the reciprocal antibody dilution that reduced the number of viral foci by 50%. IgG antibodies were first purified from each serum in order to avoid the facilitating effect of HDL on HCV entry. RESULTS: The assay's cut-off using an ELISA and RNA HCV-negative samples was found to be 1.25 log, corresponding to a dilution of 1:18. The assay was compared with a commercial HCV ELISA and exhibited specificity and sensitivity values of 100% and 96.5%, respectively, and good reproducibility (with intra-assay and inter-assay coefficients of variation of 6.7% and 12.6%, respectively). The assay did not show any cross-reactivity with anti-HIV, anti-HBs or heterophile antibody positive samples. The neutralizing antibodies titers were 2.13 log (1:134) for homologous samples from HCV genotype 2 infected patients harboring the same genotype as JFH-1 and 1.93 log (1:85) for heterologous samples from patients infected by genotypes other than type 2. These results confirm the presence of broadly cross-neutralizing antibodies already reported using the HCV pseudoparticles system. CONCLUSION: This study presents a simple, specific and reproducible cell culture-based assay for determination of HCV-neutralizing antibodies in human sera. The assay should be an important tool for gauging the relationship between the neutralizing antibodies response and viral load kinetics in acutely or chronically infected patients and for investigating the possible eradication or prevention of HCV infection by neutralizing antibodies. PMID- 17397532 TI - A meta-analysis of kidney microarray datasets: investigation of cytokine gene detection and correlation with rt-PCR and detection thresholds. AB - BACKGROUND: Microarrays provide a means to simultaneously examine the gene expression of the entire transcriptome in a single sample. Many studies have highlighted the need for novel software and statistical approaches to assess the measured gene expression. Less attention has been directed toward whether genes considered undetectable by microarray can be detected by other strategies or whether these genes can provide accurate gene expression determinations. In the kidney this is a concern for genes such as cytokines which dramatically influence the immune response but are often considered low abundance genes produced by a small number of cells. RESULTS: Using both publicly available and our own microarray datasets we analyzed the detection p-value and detection call values for 81 human kidney samples run on the U133A or U133Plus2.0 Affymetrix microarrays (Affymetrix, Santa Clara, CA). For the cytokine genes, the frequency of detection in each sample group (normal, transplant and renal cell carcinoma) was examined and revealed that a majority of cytokine related genes are not detectable in human kidney by microarray. Using a subset of 29 Mayo transplant samples, a group of seven transplant-related cytokines and eight non-cytokine genes were evaluated by real-time PCR (rt-PCR). For these 15 genes we compared the impact of decreasing microarray detection frequency with the changes in gene expression observed by both microarray and rt-PCR. We found that as microarray detection frequency decreased the correlation between microarray and rt-PCR data also decreased. CONCLUSION: We conclude that, when analyzing microarray data from human kidney samples, genes generally expressed at low abundance (i.e. cytokines) should be evaluated with more sensitive approaches such as rt-PCR. In addition, our data suggest that the use of detection frequency cutoffs for inclusion or exclusion of microarray data may be appropriate when comparing microarray and rt PCR gene expression data and p-value calculations. PMID- 17397533 TI - Morphological characterization of intra-articular HMGB1 expression during the course of collagen-induced arthritis. AB - High-mobility group chromosomal box protein 1 (HMGB1) is a structural nuclear protein that promotes inflammation when present extracellularly. Aberrant, extracellular HMGB1 expression has been demonstrated in human and experimental synovitis. The aim of the present study was to elucidate the temporal and spatial expression of HMGB1 compared to that of the central mediators tumor necrosis factor (TNF) and interleukin-1-beta (IL-1beta) during the course of collagen induced arthritis. Thus, Dark Agouti rats were immunized with homologous type II collagen and synovial tissue specimens were obtained at various time points prior to and during the course of clinical arthritis. Local cytokine responses were assessed by immunohistochemistry and by in situ hybridization. We demonstrate a distinct nuclear expression of HMGB1 at early disease-preceding time points. Preceding clinical onset by a few days, cytoplasmic HMGB1 expression was evident in synoviocytes within the non-proliferative lining layer. Pronounced cytoplasmic and additional extracellular HMGB1 expression coincided with the progression of clinical disease. In advanced arthritis, the number of cells with cytoplasmic HMGB1 expression was quantitatively comparable to that of cells expressing TNF and IL-1beta. Interestingly, although HMGB1 was abundantly expressed throughout the inflamed synovium at a protein level, upregulation of HMGB1 mRNA was restricted mainly to areas of cartilage and bone destruction. In conclusion, these new findings implicate a role for HMGB1 in both inducing and perpetuating inflammatory events of significant importance in the destructive processes in chronic arthritis. PMID- 17397534 TI - Are we under-utilizing the talents of primary care personnel? A job analytic examination. AB - BACKGROUND: Primary care staffing decisions are often made unsystematically, potentially leading to increased costs, dissatisfaction, turnover, and reduced quality of care. This article aims to (1) catalogue the domain of primary care tasks, (2) explore the complexity associated with these tasks, and (3) examine how tasks performed by different job titles differ in function and complexity, using Functional Job Analysis to develop a new tool for making evidence-based staffing decisions. METHODS: Seventy-seven primary care personnel from six US Department of Veterans Affairs (VA) Medical Centers, representing six job titles, participated in two-day focus groups to generate 243 unique task statements describing the content of VA primary care. Certified job analysts rated tasks on ten dimensions representing task complexity, skills, autonomy, and error consequence. Two hundred and twenty-four primary care personnel from the same clinics then completed a survey indicating whether they performed each task. Tasks were catalogued using an adaptation of an existing classification scheme; complexity differences were tested via analysis of variance. RESULTS: Objective one: Task statements were categorized into four functions: service delivery (65%), administrative duties (15%), logistic support (9%), and workforce management (11%). Objective two: Consistent with expectations, 80% of tasks received ratings at or below the mid-scale value on all ten scales. Objective three: Service delivery and workforce management tasks received higher ratings on eight of ten scales (multiple functional complexity dimensions, autonomy, human error consequence) than administrative and logistic support tasks. Similarly, tasks performed by more highly trained job titles received higher ratings on six of ten scales than tasks performed by lower trained job titles. Contrary to expectations, the distribution of tasks across functions did not significantly vary by job title. CONCLUSION: Primary care personnel are not being utilized to the extent of their training; most personnel perform many tasks that could reasonably be performed by personnel with less training. Primary care clinics should use evidence-based information to optimize job-person fit, adjusting clinic staff mix and allocation of work across staff to enhance efficiency and effectiveness. PMID- 17397535 TI - Muscular cystic hydatidosis: case report. AB - BACKGROUND: Hydatidosis is a zoonosis caused by Echinococcus granulosus, and ingesting eggs released through the faeces from infected dogs infects humans. The location of the hydatid cysts is mostly hepatic and/or pulmonary, whereas musculoskeletal hydatidosis is very rare. CASE PRESENTATION: We report an unusual case of primary muscular hydatidosis in proximity of the big adductor in a young Sicilian man. The patient, 34 years old, was admitted to the Department of Infectious and Tropical Diseases for ultrasonographic detection, with successive confirmation by magnetic resonance imaging, of an ovular mass (13 x 8 cm) in the big adductor of the left thigh, cyst-like, and containing several small cystic formations. Serological tests for hydatidosis gave negative results. A second drawing of blood was done 10 days after the first one and showed an increase in the antibody titer for hydatidosis. The patient was submitted to surgical excision of the lesion with perioperatory prophylaxis with albendazole. The histopathological examination of the bioptic material was not diriment in the diagnosis, therefore further tests were performed: additional serological tests for hydatidosis for the evaluation of IgE and IgG serotype (Western Blot and REAST), and molecular analysis of the excised material. These more specific serological tests gave positive results for hydatidosis, and the sequencing of the polymerase chain reaction products from the cyst evidenced E. granulosus DNA, genotype G1. Any post-surgery complications was observed during 6 following months. CONCLUSION: Cystic hydatidosis should always be considered in the differential diagnosis of any cystic mass, regardless of its location, also in epidemiological contests less suggestive of the disease. The diagnosis should be achieved by taking into consideration the clinical aspects, the epidemiology of the disease, the imaging and immunological tests but, as demonstrated in this case, without neglecting the numerous possibilities offered by new serological devices and modern day molecular biology techniques. PMID- 17397536 TI - Serum from mice immunized in the context of Treg inhibition identifies DEK as a neuroblastoma tumor antigen. AB - BACKGROUND: We have developed a cell-based vaccine that features the expression of both CD80 and CD86 on the surface of a murine neuroblastoma cell line. The cellular immunity induced by this vaccine is enhanced by treatment with antibody that interferes with T-regulatory cell (Treg) function and we report here that immunization combined with interfering with Treg function also produces a profound serological effect. Serum from mice immunized with our cell-based vaccine in the context of Treg blockade was used to screen a cDNA expression library constructed from the parental neuroblastoma tumor cell line, AGN2a. RESULTS: Serum from mice vaccinated in the context of Treg blockade identified a number of potentially oncogenic transcripts that may serve as important immune targets in a tumor-derived cDNA library screen. This novel approach identified far more candidates than could be seen with serum derived from vaccine-treated only, Treg-depleted only, or tumor-bearing mice. The most commonly identified tumor-associated antigen, using serum from immunized and Treg-depleted mice, was the DEK oncogene. Altered expression of the DEK oncogene has been implicated in a number of human cancers. Importantly, we were able to demonstrate that the DEK oncogene also induces a T cell response. CONCLUSION: The use of post-vaccine immune serum in this report differs from previous approaches where serum collected at the time of cancer onset or diagnosis and was used for tumor antigen identification. We hypothesize that the use of diagnostic serum samples may be inadequate for the clinical translation of this approach, and that identification of protective immunogenic tumor antigens may require the use of serum from post treatment or vaccinated subjects. The identification of DEK as a tumor-associated antigen capable of eliciting a T cell response validates our experimental approach and argues for the antigens we have identified here to be evaluated as targets of effector immunity and as vaccine candidates. PMID- 17397537 TI - Atomic hydration potentials using a Monte Carlo Reference State (MCRS) for protein solvation modeling. AB - BACKGROUND: Accurate description of protein interaction with aqueous solvent is crucial for modeling of protein folding, protein-protein interaction, and drug design. Efforts to build a working description of solvation, both by continuous models and by molecular dynamics, yield controversial results. Specifically constructed knowledge-based potentials appear to be promising for accounting for the solvation at the molecular level, yet have not been used for this purpose. RESULTS: We developed original knowledge-based potentials to study protein hydration at the level of atom contacts. The potentials were obtained using a new Monte Carlo reference state (MCRS), which simulates the expected probability density of atom-atom contacts via exhaustive sampling of structure space with random probes. Using the MCRS allowed us to calculate the expected atom contact densities with high resolution over a broad distance range including very short distances. Knowledge-based potentials for hydration of protein atoms of different types were obtained based on frequencies of their contacts at different distances with protein-bound water molecules, in a non-redundant training data base of 1776 proteins with known 3D structures. Protein hydration sites were predicted in a test set of 12 proteins with experimentally determined water locations. The MCRS greatly improves prediction of water locations over existing methods. In addition, the contribution of the energy of macromolecular solvation into total folding free energy was estimated, and tested in fold recognition experiments. The correct folds were preferred over all the misfolded decoys for the majority of proteins from the improved Rosetta decoy set based on the structure hydration energy alone. CONCLUSION: MCRS atomic hydration potentials provide a detailed distance-dependent description of hydropathies of individual protein atoms. This allows placement of water molecules on the surface of proteins and in protein interfaces with much higher precision. The potentials provide a means to estimate the total solvation energy for a protein structure, in many cases achieving a successful fold recognition. Possible applications of atomic hydration potentials to structure verification, protein folding and stability, and protein-protein interactions are discussed. PMID- 17397538 TI - Molecular characterisation of the STRUBBELIG-RECEPTOR FAMILY of genes encoding putative leucine-rich repeat receptor-like kinases in Arabidopsis thaliana. AB - BACKGROUND: Receptor-like kinases are a prominent class of surface receptors that regulate many aspects of the plant life cycle. Despite recent advances the function of most receptor-like kinases remains elusive. Therefore, it is paramount to investigate these receptors. The task is complicated by the fact that receptor-like kinases belong to a large monophyletic family with many sub clades. In general, functional analysis of gene family members by reverse genetics is often obscured by several issues, such as redundancy, subtle or difficult to detect phenotypes in mutants, or by decision problems regarding suitable biological and biochemical assays. Therefore, in many cases additional strategies have to be employed to allow inference of hypotheses regarding gene function. RESULTS: We approached the function of genes encoding the nine-member STRUBBELIG-RECEPTOR FAMILY (SRF) class of putative leucine-rich repeat receptor like kinases. Sequence comparisons show overall conservation but also divergence in predicted functional domains among SRF proteins. Interestingly, SRF1 undergoes differential splicing. As a result, SRF1 is predicted to exist in a standard receptor configuration and in a membrane-anchored receptor-like version that lacks most of the intracellular domain. Furthermore, SRF1 is characterised by a high degree of polymorphism between the Ler and Col accessions. Two independent T DNA-based srf4 mutants showed smaller leaves while 35S::SRF4 plants displayed enlarged leaves. This is in addition to the strubbelig phenotype which has been described before. Additional single and several key double mutant combinations did not reveal obvious mutant phenotypes. Ectopic expression of several SRF genes, using the 35S promoter, resulted in male sterility. To gain possible insights into SRF gene function we employed a computational analysis of publicly available microarray data. We performed global expression profiling, coexpression analysis, and an analysis of the enrichment of gene ontology terms among coexpressed genes. The bioinformatic analyses raise the possibility that some SRF genes affect different aspects of cell wall biology. The results also indicate that redundancy is a minor aspect of the SRF family. CONCLUSION: The results provide evidence that SRF4 is a positive regulator of leaf size. In addition, they suggest that the SRF family is characterised by functional diversity and that some SRF genes may function in cell wall biology. They also indicate that complementing reverse genetics with bioinformatical data mining of genome-wide expression data aids in inferring hypotheses on possible functions for members of a gene family. PMID- 17397539 TI - Transcriptional regulatory network discovery via multiple method integration: application to e. coli K12. AB - Transcriptional regulatory network (TRN) discovery from one method (e.g. microarray analysis, gene ontology, phylogenic similarity) does not seem feasible due to lack of sufficient information, resulting in the construction of spurious or incomplete TRNs. We develop a methodology, TRND, that integrates a preliminary TRN, microarray data, gene ontology and phylogenic similarity to accurately discover TRNs and apply the method to E. coli K12. The approach can easily be extended to include other methodologies. Although gene ontology and phylogenic similarity have been used in the context of gene-gene networks, we show that more information can be extracted when gene-gene scores are transformed to gene transcription factor (TF) scores using a preliminary TRN. This seems to be preferable over the construction of gene-gene interaction networks in light of the observed fact that gene expression and activity of a TF made of a component encoded by that gene is often out of phase. TRND multi-method integration is found to be facilitated by the use of a Bayesian framework for each method derived from its individual scoring measure and a training set of gene/TF regulatory interactions. The TRNs we construct are in better agreement with microarray data. The number of gene/TF interactions we discover is actually double that of existing networks. PMID- 17397540 TI - Spatial mosaic evolution of snail defensive traits. AB - BACKGROUND: Recent models suggest that escalating reciprocal selection among antagonistically interacting species is predicted to occur in areas of higher resource productivity. In a putatively coevolved interaction between a freshwater snail (Mexipyrgus churinceanus) and a molluscivorous cichlid (Herichthys minckleyi), we examined three components of this interaction: 1) spatial variation in two putative defensive traits, crushing resistance and shell pigmentation; 2) whether abiotic variables or frequency of molariform cichlids are associated with spatial patterns of crushing resistance and shell pigmentation and 3) whether variation in primary productivity accounted for small scale variation in these defensive traits. RESULTS: Using spatial autocorrelation to account for genetic and geographic divergence among populations, we found no autocorrelation among populations at small geographic and genetic distances for the two defensive traits. There was also no correlation between abiotic variables (temperature and conductivity) and snail defensive traits. However, crushing resistance and frequency of pigmented shells were negatively correlated with molariform frequency. Crushing resistance and levels of pigmentation were significantly higher in habitats dominated by aquatic macrophytes, and both traits are phenotypically correlated. CONCLUSION: Crushing resistance and pigmentation of M. churinceanus exhibit striking variation at small spatial scales often associated with differences in primary productivity, substrate coloration and the frequency of molariform cichlids. These local geographic differences may result from among-habitat variation in how resource productivity interacts to promote escalation in prey defenses. PMID- 17397541 TI - Structure, substrate recognition and reactivity of Leishmania major mevalonate kinase. AB - BACKGROUND: Isoprenoid precursor synthesis via the mevalonate route in humans and pathogenic trypanosomatids is an important metabolic pathway. There is however, only limited information available on the structure and reactivity of the component enzymes in trypanosomatids. Since isoprenoid biosynthesis is essential for trypanosomatid viability and may provide new targets for therapeutic intervention it is important to characterize the pathway components. RESULTS: Putative mevalonate kinase encoding genes from Leishmania major (LmMK) and Trypanosoma brucei (TbMK) have been cloned, over-expressed in and proteins isolated from procyclic-form T. brucei. A highly sensitive radioactive assay was developed and shows ATP-dependent phosphorylation of mevalonate. Apo and (R) mevalonate bound crystal structures of LmMK, from a bacterial expression system, have been determined to high resolution providing, for the first time, information concerning binding of mevalonate to an MK. The mevalonate binds in a deep cavity lined by highly conserved residues. His25 is key for binding and for discrimination of (R)- over (S)-mevalonate, with the main chain amide interacting with the C3 hydroxyl group of (R)-mevalonate, and the side chain contributing, together with Val202 and Thr283, to the construction of a hydrophobic binding site for the C3 methyl substituent. The C5 hydroxyl, where phosphorylation occurs, points towards catalytic residues, Lys18 and Asp155. The activity of LmMK was significantly reduced compared to MK from other species and we were unable to obtain ATP-binding data. Comparisons with the rat MK:ATP complex were used to investigate how this substrate might bind. In LmMK, helix alpha2 and the preceding polypeptide adopt a conformation, not seen in related kinase structures, impeding access to the nucleotide triphosphate binding site suggesting that a conformational rearrangement is required to allow ATP binding. CONCLUSION: Our new structural information, consistent with data on homologous enzymes allows a detailed description of how mevalonate is recognized and positioned for catalysis in MK. The mevalonate-binding site is highly conserved yet the ATP-binding site is structurally distinct in LmMK. We are unable to provide a definitive explanation for the low activity of recombinant protein isolated from a bacterial expression system compared to material isolated from procyclic-form Trypanosoma brucei. PMID- 17397542 TI - Trends in complementary/alternative medicine use by breast cancer survivors: comparing survey data from 1998 and 2005. AB - BACKGROUND: Use of complementary and alternative medicine (CAM) by women with breast cancer is often said to be increasing, yet few data exist to confirm this commonly held belief. The purpose of this paper is to compare overall patterns of CAM use, as well as use of specific products and therapies at two different points in time (1998 vs 2005) by women diagnosed with breast cancer. METHODS: Surveys were mailed to women randomly selected from the Ontario Cancer Registry (Canada) in the spring of 1998 (n = 557) and again in the spring of 2005(n = 877). RESULTS: The response rates were 76.3% in 1998 and 63% in 2005. In 1998, 66.7% of women reported using either a CAM product/therapy or seeing a CAM therapist at some time in their lives as compared with 81.9% in 2005 (p = 0.0002). Increases were seen in both use of CAM products/therapies (62% in 1998 vs. 70.6% in 2005) and visits to CAM practitioners (39.4% of respondents in 1998 vs 57.4% of respondents in 2005). Women in 2005 reported that 41% used CAM for treating their breast cancer. The most commonly used products and practitioners for treating breast cancer as reported in 2005 were green tea, vitamin E, flaxseed, vitamin C, massage therapists and dietitians/nutritionists. CONCLUSION: CAM use (both self-medication with products and visits to CAM practitioners) increased significantly from 1998 to 2005. Now that more than 80% of all women with breast cancer report using CAM (41% in a specific attempt to management their breast cancer), CAM use can no longer be regarded as an "alternative" or unusual approach to managing breast cancer. PMID- 17397544 TI - Exploring students' perceptions on the use of significant event analysis, as part of a portfolio assessment process in general practice, as a tool for learning how to use reflection in learning. AB - BACKGROUND: Portfolio learning enables students to collect evidence of their learning. Component tasks making up a portfolio can be devised that relate directly to intended learning outcomes. Reflective tasks can stimulate students to recognise their own learning needs. Assessment of portfolios using a rating scale relating to intended learning outcomes offers high content validity. This study evaluated a reflective portfolio used during a final-year attachment in general practice (family medicine). Students were asked to evaluate the portfolio (which used significant event analysis as a basis for reflection) as a learning tool. The validity and reliability of the portfolio as an assessment tool were also measured. METHODS: 81 final-year medical students completed reflective significant event analyses as part of a portfolio created during a three-week attachment (clerkship) in general practice (family medicine). As well as two reflective significant event analyses each portfolio contained an audit and a health needs assessment. Portfolios were marked three times; by the student's GP teacher, the course organiser and by another teacher in the university department of general practice. Inter-rater reliability between pairs of markers was calculated. A questionnaire enabled the students' experience of portfolio learning to be determined. RESULTS: Benefits to learning from reflective learning were limited. Students said that they thought more about the patients they wrote up in significant event analyses but information as to the nature and effect of this was not forthcoming. Moderate inter-rater reliability (Spearman's Rho .65) was found between pairs of departmental raters dealing with larger numbers (20 60) of portfolios. Inter-rater reliability of marking involving GP tutors who only marked 1-3 portfolios was very low. Students rated highly their mentoring relationship with their GP teacher but found the portfolio tasks time-consuming. CONCLUSION: The inter-rater reliability observed in this study should be viewed alongside the high validity afforded by the authenticity of the learning tasks (compared with a sample of a student's learning taken by an exam question). Validity is enhanced by the rating scale which directly connects the grade given with intended learning outcomes. The moderate inter-rater reliability may be increased if a portfolio is completed over a longer period of time and contains more component pieces of work. The questionnaire used in this study only accessed limited information about the effect of reflection on students' learning. Qualitative methods of evaluation would determine the students experience in greater depth. It would be useful to evaluate the effects of reflective learning after students have had more time to get used to this unfamiliar method of learning and to overcome any problems in understanding the task. PMID- 17397543 TI - Vitamin D and the vitamin D receptor are critical for control of the innate immune response to colonic injury. AB - BACKGROUND: The active form of vitamin D (1,25(OH)2D3) has been shown to inhibit development of inflammatory bowel disease (IBD) in IL-10 KO mice. Here, the role of the vitamin D receptor (VDR) and 1,25(OH)2D3 in acute experimental IBD was probed. RESULTS: VDR KO mice were extremely sensitive to dextran sodium sulfate (DSS) and there was increased mortality of the VDR KO mice at doses of DSS that only caused a mild form of colitis in wildtype (WT) mice. DSS colitis in the VDR KO mice was accompanied by high colonic expression of TNF-alpha, IL-1 alpha, IL 1beta, IL-12, IFN-gamma, IL-10, MIP-1alpha and KC. DSS concentrations as low as 0.5% were enough to induce bleeding, ulceration and weight loss in VDR KO mice. VDR KO mice failed to recover following the removal of DSS, while WT mice showed signs of recovery within 5 days of DSS removal. The early mortality of DSS treated VDR KO mice was likely due to perforation of the bowel and resulting endotoxemia. VDR KO mice were hyper-responsive to exogenously injected LPS and cultures of the peritoneal exudates of moribund DSS treated VDR KO mice were positive for bacterial growth. 1,25(OH)2D3 in the diet or rectally decreased the severity and extent of DSS-induced inflammation in WT mice. CONCLUSION: The data point to a critical role for the VDR and 1,25(OH)2D3 in control of innate immunity and the response of the colon to chemical injury. PMID- 17397545 TI - The uncoupling protein 1 gene, UCP1, is expressed in mammalian islet cells and associated with acute insulin response to glucose in African American families from the IRAS Family Study. AB - BACKGROUND: Variants of uncoupling protein genes UCP1 and UCP2 have been associated with a range of traits. We wished to evaluate contributions of known UCP1 and UCP2 variants to metabolic traits in the Insulin Resistance and Atherosclerosis (IRAS) Family Study. METHODS: We genotyped five promoter or coding single nucleotide polymorphisms (SNPs) in 239 African American (AA) participants and 583 Hispanic participants from San Antonio (SA) and San Luis Valley. Generalized estimating equations using a sandwich estimator of the variance and exchangeable correlation to account for familial correlation were computed for the test of genotypic association, and dominant, additive and recessive models. Tests were adjusted for age, gender and BMI (glucose homeostasis and lipid traits), or age and gender (obesity traits), and empirical P-values estimated using a gene dropping approach. RESULTS: UCP1 A-3826G was associated with AIR(g) in AA (P = 0.006) and approached significance in Hispanic families (P = 0.054); and with HDL-C levels in SA families (P = 0.0004). Although UCP1 expression is reported to be restricted to adipose tissue, RT-PCR indicated that UCP1 is expressed in human pancreas and MIN-6 cells, and immunohistochemistry demonstrated co-localization of UCP1 protein with insulin in human islets. UCP2 A55V was associated with waist circumference (P = 0.045) in AA, and BMI in SA (P = 0.018); and UCP2 G-866A with waist-to-hip ratio in AA (P = 0.016). CONCLUSION: This study suggests a functional variant of UCP1 contributes to the variance of AIR(g) in an AA population; the plausibility of this unexpected association is supported by the novel finding that UCP1 is expressed in islets. PMID- 17397546 TI - Telephone survey of private patients' views on continuity of care and registration with general practice in Ireland. AB - BACKGROUND: The desire of patients for personal continuity of care with a General Practitioner (GP) has been well documented, but not within non-registered private patients in Ireland. This study set out to examine the attitudes and reported behaviours of private fee-paying patients towards continuity of GP care and universal registration for patients. METHODS: Cross-sectional telephone survey of 400 randomly chosen fee-paying patients living within County Dublin. There is no formal system of registration with a GP for these patients. Main outcomes were attendance of respondents at primary health care facilities and their attitudes towards continuity of care and registration with a GP. Data was analysed using descriptive statistics and using parametric and non-parametric tests of association. Pearson correlation was used to quantify the association between the described variables and attitudes towards continuity and registration with a GP. Variables showing significance at the 5% level were entered into multiple linear regression models. RESULTS: 97% of respondents had seen a GP in the previous 5 years. The mean number of visits to the GP for respondents was 2.3 per annum. 89% of respondents had a regular GP and the mean length of time with their GP was 15.6 years. 96% preferred their personal medical care to be provided within one general practice. 16% of respondents had consulted a GP outside of their own practice in the previous year. They were more likely to be female, commute a longer distance to work or have poorer health status. 81% considered it important to be officially registered with a GP practice of their choice. CONCLUSION: Both personal and longitudinal continuity of care with a GP are important to private patients. Respondents who chose to visit GPs other than their regular GP were not easily characterised in this study and individual circumstances may lead to this behaviour. There is strong support for a system of universal patient registration within general practice. PMID- 17397547 TI - Regulatory network of inflammation downstream of proteinase-activated receptors. AB - BACKGROUND: Protease-activated receptors (PAR) are present in the urinary bladder, and their expression is altered in response to inflammation. PARs are a unique class of G protein-coupled that carry their own ligands, which remain cryptic until unmasked by proteolytic cleavage. Although the canonical signal transduction pathway downstream of PAR activation and coupling with various G proteins is known and leads to the rapid transcription of genes involved in inflammation, the effect of PAR activation on the downstream transcriptome is unknown. We have shown that intravesical administration of PAR-activating peptides leads to an inflammatory reaction characterized by edema and granulocyte infiltration. Moreover, the inflammatory response to intravesical instillation of known pro-inflammatory stimuli such as E. coli lipopolysaccharide (LPS), substance P (SP), and antigen was strongly attenuated by PAR1- and to a lesser extent by PAR2-deficiency. RESULTS: Here, cDNA array experiments determined inflammatory genes whose expression is dependent on PAR1 activation. For this purpose, we compared the alteration in gene expression in wild type and PAR1-/- mice induced by classical pro-inflammatory stimuli (LPS, SP, and antigen). 75 transcripts were considered to be dependent on PAR-1 activation and further annotated in silico by Ingenuity Pathways Analysis (IPA) and gene ontology (GO). Selected transcripts were target validated by quantitative PCR (Q-PCR). Among PAR1-dependent transcripts, the following have been implicated in the inflammatory process: b2m, ccl7, cd200, cd63, cdbpd, cfl1, dusp1, fkbp1a, fth1, hspb1, marcksl1, mmp2, myo5a, nfkbia, pax1, plaur, ppia, ptpn1, ptprcap, s100a10, sim2, and tnfaip2. However, a balanced response to signals of injury requires a transient cellular activation of a panel of genes together with inhibitory systems that temper the overwhelming inflammation. In this context, the activation of genes such as dusp1 and nfkbia seems to counter-balance the inflammatory response to PAR activation by limiting prolonged activation of p38 MAPK and increased cytokine production. In contrast, transcripts such as arf6 and dcnt1 that are involved in the mechanism of PAR re-sensitization would tend to perpetuate the inflammatory reaction in response to common pro-inflammatory stimuli. CONCLUSION: The combination of cDNA array results and genomic networks reveals an overriding participation of PAR1 in bladder inflammation, provides a working model for the involvement of downstream signaling, and evokes testable hypotheses regarding the transcriptome downstream of PAR1 activation. It remains to be determined whether or not mechanisms targeting PAR1 gene silencing or PAR1 blockade will ameliorate the clinical manifestation of cystitis. PMID- 17397549 TI - A new paradigm for depression in new mothers: the central role of inflammation and how breastfeeding and anti-inflammatory treatments protect maternal mental health. AB - BACKGROUND: Research in the field of psychoneuroimmunology (PNI) has revealed that depression is associated with inflammation manifested by increased levels of proinflammatory cytokines. DISCUSSION: The old paradigm described inflammation as simply one of many risk factors for depression. The new paradigm is based on more recent research that has indicated that physical and psychological stressors increase inflammation. These recent studies constitute an important shift in the depression paradigm: inflammation is not simply a risk factor; it is the risk factor that underlies all the others. Moreover, inflammation explains why psychosocial, behavioral and physical risk factors increase the risk of depression. This is true for depression in general and for postpartum depression in particular. Puerperal women are especially vulnerable to these effects because their levels of proinflammatory cytokines significantly increase during the last trimester of pregnancy--a time when they are also at high risk for depression. Moreover, common experiences of new motherhood, such as sleep disturbance, postpartum pain, and past or current psychological trauma, act as stressors that cause proinflammatory cytokine levels to rise. Breastfeeding has a protective effect on maternal mental health because it attenuates stress and modulates the inflammatory response. However, breastfeeding difficulties, such as nipple pain, can increase the risk of depression and must be addressed promptly. CONCLUSION: PNI research suggests two goals for the prevention and treatment of postpartum depression: reducing maternal stress and reducing inflammation. Breastfeeding and exercise reduce maternal stress and are protective of maternal mood. In addition, most current treatments for depression are anti-inflammatory. These include long chain omega-3 fatty acids, cognitive therapy, St. John's wort, and conventional antidepressants. PMID- 17397548 TI - Mandatory role of proteinase-activated receptor 1 in experimental bladder inflammation. AB - BACKGROUND: In general, inflammation plays a role in most bladder pathologies and represents a defense reaction to injury that often times is two edged. In particular, bladder neurogenic inflammation involves the participation of mast cells and sensory nerves. Increased mast cell numbers and tryptase release represent one of the prevalent etiologic theories for interstitial cystitis and other urinary bladder inflammatory conditions. The activity of mast cell-derived tryptase as well as thrombin is significantly increased during inflammation. Those enzymes activate specific G-protein coupled proteinase-activated receptors (PAR)s. Four PARs have been cloned so far, and not only are all four receptors highly expressed in different cell types of the mouse urinary bladder, but their expression is altered during experimental bladder inflammation. We hypothesize that PARs may link mast cell-derived proteases to bladder inflammation and, therefore, play a fundamental role in the pathogenesis of cystitis. RESULTS: Here, we demonstrate that in addition to the mouse urinary bladder, all four PA receptors are also expressed in the J82 human urothelial cell line. Intravesical administration of PAR-activating peptides in mice leads to an inflammatory reaction characterized by edema and granulocyte infiltration. Moreover, the inflammatory response to intravesical instillation of known pro-inflammatory stimuli such as E. coli lipopolysaccharide (LPS), substance P, and antigen was strongly attenuated by PAR1-, and to a lesser extent, by PAR2-deficiency. CONCLUSION: Our results reveal an overriding participation of PAR1 in bladder inflammation, provide a working model for the involvement of downstream signaling, and evoke testable hypotheses regarding the role of PARs in bladder inflammation. It remains to be determined whether or not mechanisms targeting PAR1 gene silencing or PAR1 blockade will ameliorate the clinical manifestations of cystitis. PMID- 17397550 TI - Clinical and medication profiles stratified by household income in patients referred for diabetes care. AB - BACKGROUND: Low income individuals with diabetes are at particularly high risk for poor health outcomes. While specialized diabetes care may help reduce this risk, it is not currently known whether there are significant clinical differences across income groups at the time of referral. The objective of this study is to determine if the clinical profiles and medication use of patients referred for diabetes care differ across income quintiles. METHODS: This cross sectional study was conducted using a Canadian, urban, Diabetes Education Centre (DEC) database. Clinical information on the 4687 patients referred to the DEC from May 2000 - January 2002 was examined. These data were merged with 2001 Canadian census data on income. Potential differences in continuous clinical parameters across income quintiles were examined using regression models. Differences in medication use were examined using Chi square analyses. RESULTS: Multivariate regression analysis indicated that income was negatively associated with BMI (p < 0.0005) and age (p = 0.023) at time of referral. The highest income quintiles were found to have lower serum triglycerides (p = 0.011) and higher HDL c (p = 0.008) at time of referral. No significant differences were found in HBA1C, LDL-c or duration of diabetes. The Chi square analysis of medication use revealed that despite no significant differences in HBA1C, the lowest income quintiles used more metformin (p = 0.001) and sulfonylureas (p < 0.0005) than the wealthy. Use of other therapies were similar across income groups, including lipid lowering medications. High income patients were more likely to be treated with diet alone (p < 0.0005). CONCLUSION: Our findings demonstrate that low income patients present to diabetes clinic older, heavier and with a more atherogenic lipid profile than do high income patients. Overall medication use was higher among the lower income group suggesting that differences in clinical profiles are not the result of under-treatment, thus invoking lifestyle factors as potential contributors to these findings. PMID- 17397551 TI - Conifer R2R3-MYB transcription factors: sequence analyses and gene expression in wood-forming tissues of white spruce (Picea glauca). AB - BACKGROUND: Several members of the R2R3-MYB family of transcription factors act as regulators of lignin and phenylpropanoid metabolism during wood formation in angiosperm and gymnosperm plants. The angiosperm Arabidopsis has over one hundred R2R3-MYBs genes; however, only a few members of this family have been discovered in gymnosperms. RESULTS: We isolated and characterised full-length cDNAs encoding R2R3-MYB genes from the gymnosperms white spruce, Picea glauca (13 sequences), and loblolly pine, Pinus taeda L. (five sequences). Sequence similarities and phylogenetic analyses placed the spruce and pine sequences in diverse subgroups of the large R2R3-MYB family, although several of the sequences clustered closely together. We searched the highly variable C-terminal region of diverse plant MYBs for conserved amino acid sequences and identified 20 motifs in the spruce MYBs, nine of which have not previously been reported and three of which are specific to conifers. The number and length of the introns in spruce MYB genes varied significantly, but their positions were well conserved relative to angiosperm MYB genes. Quantitative RTPCR of MYB genes transcript abundance in root and stem tissues revealed diverse expression patterns; three MYB genes were preferentially expressed in secondary xylem, whereas others were preferentially expressed in phloem or were ubiquitous. The MYB genes expressed in xylem, and three others, were up-regulated in the compression wood of leaning trees within 76 hours of induction. CONCLUSION: Our survey of 18 conifer R2R3-MYB genes clearly showed a gene family structure similar to that of Arabidopsis. Three of the sequences are likely to play a role in lignin metabolism and/or wood formation in gymnosperm trees, including a close homolog of the loblolly pine PtMYB4, shown to regulate lignin biosynthesis in transgenic tobacco. PMID- 17397552 TI - Locational distribution of gene functional classes in Arabidopsis thaliana. AB - BACKGROUND: We are interested in understanding the locational distribution of genes and their functions in genomes, as this distribution has both functional and evolutionary significance. Gene locational distribution is known to be affected by various evolutionary processes, with tandem duplication thought to be the main process producing clustering of homologous sequences. Recent research has found clustering of protein structural families in the human genome, even when genes identified as tandem duplicates have been removed from the data. However, this previous research was hindered as they were unable to analyse small sample sizes. This is a challenge for bioinformatics as more specific functional classes have fewer examples and conventional statistical analyses of these small data sets often produces unsatisfactory results. RESULTS: We have developed a novel bioinformatics method based on Monte Carlo methods and Greenwood's spacing statistic for the computational analysis of the distribution of individual functional classes of genes (from GO). We used this to make the first comprehensive statistical analysis of the relationship between gene functional class and location on a genome. Analysis of the distribution of all genes except tandem duplicates on the five chromosomes of A. thaliana reveals that the distribution on chromosomes I, II, IV and V is clustered at P = 0.001. Many functional classes are clustered, with the degree of clustering within an individual class generally consistent across all five chromosomes. A novel and surprising result was that the locational distribution of some functional classes were significantly more evenly spaced than would be expected by chance. CONCLUSION: Analysis of the A. thaliana genome reveals evidence of unexplained order in the locational distribution of genes. The same general analysis method can be applied to any genome, and indeed any sequential data involving classes. PMID- 17397553 TI - Tandem repeat regions within the Burkholderia pseudomallei genome and their application for high resolution genotyping. AB - BACKGROUND: The facultative, intracellular bacterium Burkholderia pseudomallei is the causative agent of melioidosis, a serious infectious disease of humans and animals. We identified and categorized tandem repeat arrays and their distribution throughout the genome of B. pseudomallei strain K96243 in order to develop a genetic typing method for B. pseudomallei. We then screened 104 of the potentially polymorphic loci across a diverse panel of 31 isolates including B. pseudomallei, B. mallei and B. thailandensis in order to identify loci with varying degrees of polymorphism. A subset of these tandem repeat arrays were subsequently developed into a multiple-locus VNTR analysis to examine 66 B. pseudomallei and 21 B. mallei isolates from around the world, as well as 95 lineages from a serial transfer experiment encompassing ~18,000 generations. RESULTS: B. pseudomallei contains a preponderance of tandem repeat loci throughout its genome, many of which are duplicated elsewhere in the genome. The majority of these loci are composed of repeat motif lengths of 6 to 9 bp with 4 to 10 repeat units and are predominately located in intergenic regions of the genome. Across geographically diverse B. pseudomallei and B.mallei isolates, the 32 VNTR loci displayed between 7 and 28 alleles, with Nei's diversity values ranging from 0.47 and 0.94. Mutation rates for these loci are comparable (>10-5 per locus per generation) to that of the most diverse tandemly repeated regions found in other less diverse bacteria. CONCLUSION: The frequency, location and duplicate nature of tandemly repeated regions within the B. pseudomallei genome indicate that these tandem repeat regions may play a role in generating and maintaining adaptive genomic variation. Multiple-locus VNTR analysis revealed extensive diversity within the global isolate set containing B. pseudomallei and B. mallei, and it detected genotypic differences within clonal lineages of both species that were identical using previous typing methods. Given the health threat to humans and livestock and the potential for B. pseudomallei to be released intentionally, MLVA could prove to be an important tool for fine-scale epidemiological or forensic tracking of this increasingly important environmental pathogen. PMID- 17397554 TI - Comparison of the systemic and pulmonary inflammatory response to endotoxin of neutropenic and non-neutropenic rats. AB - BACKGROUND: Neutrophil infiltration commonly occurs in acute lung injury and may be partly responsible for the inflammatory response. However, acute lung injury still occurs in the neutropenic host. The objectives of this study are to determine if inflammation and acute lung injury are worse in neutropenic versus the normal host after endotoxemia. METHODS: Rats were divided into four groups: 1) control, 2) neutropenic, 3) endotoxemic and 4) endotoxemic and neutropenic. Tumor necrosis factor (TNF)-alpha and macrophage inflammatory protein (MIP-2) were measured in the blood, lung lavage and for mRNA in the lung. Arterial blood gases were measured to determine the alveolar-arterial oxygen gradient which reflects on lung injury. RESULTS: In endotoxemia, the neutropenic rats had lower plasma TNF-alpha (116 +/- 73 vs. 202 +/- 31 pg/ml) and higher plasma MIP-2 (26.8 + 11.9 vs. 15.6 + 6.9 ng/ml) when compared to non-neutropenic rats. The endotoxemic, neutropenic rats had worse lung injury than the endotoxemic, non neutropenic rats as shown by increase in the alveolar-arterial oxygen gradient (24 +/- 5 vs. 12 +/- 9 torr). However, lavage concentrations of TNF-alpha and MIP 2 were similar in both groups. CONCLUSION: Neutrophils may regulate TNF-alpha and MIP-2 production in endotoxemia. The elevation in plasma MIP-2 in the endotoxemic, neutropenic rat may be secondary to the lack of a neutrophil response to inhibit production or release of MIP-2. In endotoxemia, the severe lung injury observed in neutropenic rats does not depend on TNF-alpha or MIP-2 produced in the lung. PMID- 17397555 TI - Investigation of SNPs in the porcine desmoglein 1 gene. AB - BACKGROUND: Desmoglein 1 (DSG1) is the target protein in the skin disease exudative epidermitis in pigs caused by virulent strains of Staphylococcus hyicus. The exfoliative toxins produced by S. hyicus digest the porcine desmoglein 1 (PIG)DSG1 by a very specific reaction. This study investigated the location of single nucleotide polymorphisms (SNPs) in the porcine desmoglein 1 gene (PIG)DSG1 in correlation to the cleavage site as well as if the genotype of the SNPs is correlated to susceptibility or resistance to the disease. RESULTS: DNA from 32 affected and 32 unaffected piglets with exudative epidermitis were diagnosed clinically as affected or unaffected. Two regions of the desmoglein 1 gene were sequenced and genotypes of the SNPs were established. Seven SNPs (823T>C, 828A>G, 829A>G, 830A>T, 831A>T, 838A>C and 1139C>T) were found in the analysed sequences and the allele frequencies were determined for the SNPs resulting in amino acid change. Four of the seven polymorphisms were situated in the motif known to be important for toxin cleavage. The distribution of the genotypes between affected and unaffected animals was analysed. CONCLUSION: The study indicated a possible correlation between the genotypes of two out of seven SNPs found in the porcine desmoglein 1 gene and the susceptibility to exudative epidermitis. PMID- 17397556 TI - Intramuscular Olanzapine - a UK case series of early cases. AB - BACKGROUND: Clinical trials assessing efficacy and safety of Intramuscular (IM) Olanzapine in acute schizophrenia and acute mania have previously been undertaken in studies required for drug registration in patients who were required to give informed consent. These patients may have less severe forms of psychosis than patients treated in routine practice. Data derived from naturalistic practice following the launch of IM olanzapine may be helpful for clinicians in assessing efficacy and safety of IM olanzapine. The PANSS-EC scale used in the clinical studies may represent a tool that could be used in routine clinical practice. CASE PRESENTATION: We report on an early unselected case series of 7 patients who received IM olanzapine in routine clinical practice settings in the UK. In this case series, olanzapine IM was generally effective, and no adverse events were reported. Adjunctive benzodiazepines were given concomitantly in 1 of the 7 subjects. This is relevant as concomitant benzodiazepines are not recommended for a minimum of 1 hour post IM olanzapine administration. PANSS-EC data was collected in 2 of the 7 subjects. CONCLUSION: Although patients had greater severity of psychosis than clinical trial patients there were no unexpected findings. In addition the PANSS-EC scale is a scale that may be useful in assessing the efficacy of IM antipsychotics in routine clinical practice. PMID- 17397557 TI - Analysis of meiotic recombination in 22q11.2, a region that frequently undergoes deletions and duplications. AB - BACKGROUND: The 22q11.2 deletion syndrome is the most frequent genomic disorder with an estimated frequency of 1/4000 live births. The majority of patients (90%) have the same deletion of 3 Mb (Typically Deleted Region, TDR) that results from aberrant recombination at meiosis between region specific low-copy repeats (LCRs). METHODS: As a first step towards the characterization of recombination rates and breakpoints within the 22q11.2 region we have constructed a high resolution recombination breakpoint map based on pedigree analysis and a population-based historical recombination map based on LD analysis. RESULTS: Our pedigree map allows the location of recombination breakpoints with a high resolution (potential recombination hotspots), and this approach has led to the identification of 5 breakpoint segments of 50 kb or less (8.6 kb the smallest), that coincide with historical hotspots. It has been suggested that aberrant recombination leading to deletion (and duplication) is caused by low rates of Allelic Homologous Recombination (AHR) within the affected region. However, recombination rate estimates for 22q11.2 region show that neither average recombination rates in the 22q11.2 region or within LCR22-2 (the LCR implicated in most deletions and duplications), are significantly below chromosome 22 averages. Furthermore, LCR22-2, the repeat most frequently implicated in rearrangements, is also the LCR22 with the highest levels of AHR. In addition, we find recombination events in the 22q11.2 region to cluster within families. Within this context, the same chromosome recombines twice in one family; first by AHR and in the next generation by NAHR resulting in an individual affected with the del22q11.2 syndrome. CONCLUSION: We show in the context of a first high resolution pedigree map of the 22q11.2 region that NAHR within LCR22 leading to duplications and deletions cannot be explained exclusively under a hypothesis of low AHR rates. In addition, we find that AHR recombination events cluster within families. If normal and aberrant recombination are mechanistically related, the fact that LCR22s undergo frequent AHR and that we find familial differences in recombination rates within the 22q11.2 region would have obvious health-related implications. PMID- 17397558 TI - Demographic and occupational predictors of early response to a mailed invitation to enroll in a longitudinal health study. AB - BACKGROUND: Often in survey research, subsets of the population invited to complete the survey do not respond in a timely manner and valuable resources are expended in recontact efforts. Various methods of improving response have been offered, such as reducing questionnaire length, offering incentives, and utilizing reminders; however, these methods can be costly. Utilizing characteristics of early responders (refusal or consent) in enrollment and recontact efforts may be a unique and cost-effective approach for improving the quality of epidemiologic research. METHODS: To better understand early responders of any kind, we compared the characteristics of individuals who explicitly refused, consented, or did not respond within 2 months from the start of enrollment into a large cohort study of US military personnel. A multivariate polychotomous logistic regression model was used to estimate the effect of each covariate on the odds of early refusal and on the odds of early consent versus late/non-response, while simultaneously adjusting for all other variables in the model. RESULTS: From regression analyses, we found many similarities between early refusers and early consenters. Factors associated with both early refusal and early consent included older age, higher education, White race/ethnicity, Reserve/Guard affiliation, and certain information technology and support occupations. CONCLUSION: These data suggest that early refusers may differ from late/non-responders, and that certain characteristics are associated with both early refusal and early consent to participate. Structured recruitment efforts that utilize these differences may achieve early response, thereby reducing mail costs and the use of valuable resources in subsequent contact efforts. PMID- 17397559 TI - Empirical evaluation of the ability to learn a calorie counting system and estimate portion size and food intake. AB - The aim of this study was to determine if: (1) participants could learn the HMR Calorie System by testing if their use of the system was more accurate after training; and (2) estimated portion size and food intake improved with training. A secondary aim was to use PACE (photographic assessment of calorie estimation) to assess if participants learned the HMR system. The PACE consists of pictures of foods, the energy content of which is known. A within-subjects design was used to test the aims of this study. Participants were 44 overweight (25 or = 60 year old); 56.5% were due to nosocomial bacteremia; 46% of all patients had an underlying condition (diabetes mellitus being the most frequent [28%]); a risk factor for bacteraemia was identified in 98% (intravenous catheter: 72%). Mortality rate was 16% (21/131). Comparative analysis according to nosocomial or community acquisition showed that the former was more frequent in Surgical Wards, patients with intravenous or urinary catheters, traumatic patients, dialysis and ICU patients. Initial severity and mortality were highest in community-acquired SAMSB. In multivariate analysis, risk factors associated with higher mortality were: presence of an ultimately or rapidly fatal underlying disease, acute severity of illness at onset and inadequate empirical treatment. CONCLUSIONS: SAMSB is a clinically severe infection that occurs in patients with baseline disease, especially diabetics, that is almost always related to predisposing risk factors (mainly intravenous catheters). In the presence of a serious baseline disease, the initial critical clinical condition and inadequate empirical treatment are the main prognostic factors associated to greater mortality. PMID- 17397564 TI - [Prevalence of metabolic syndrome in the province of Albacete (Spain)]. AB - OBJECTIVE: The primary objective of this study was to determine the prevalence of Metabolic Syndrome (MS) in people between 40-70 years of age in the province of Albacete (Spain). PATIENTS AND METHODS: A population-based, cross-sectional study was made of people between 40-70 years of age in three representative municipalities of the province of Albacete. A total of 425 subjects were included, with a mean age of 53.1 years (95% CI: 52.3-54). Women represented 50.4% of the series and males 49.6%. All participants were subjected to general laboratory testing, physical examination and the measurement of anthropometric parameters. MS was defined according to the ATP-III criteria. Prevalence of MS and its distribution according to the different epidemiological characteristics were calculated. RESULTS: Total prevalence of MS was 20.9% (88/421), with a mean age of 57 years (95% CI: 55.1-59). Prevalence was seen to increase with age, reaching up to one-third of all subjects over 60 years. Significant differences were observed in relationship to a background of ischemic heart disease, ultrasensitive C-reactive protein elevation and the detection of microalbuninuria in MS subjects. Arterial hypertension and abdominal obesity were the most prevalent criteria in MS subjects. CONCLUSIONS: Taking into consideration the important co-morbidity of MS, knowledge of the prevalence and characteristics of the syndrome in our setting and its early identification and intervention targeted to the different factors underlying MS will contribute to reduce the number of cardiovascular events associated with the syndrome. PMID- 17397565 TI - [New definition of metabolic syndrome: does it have the same cardiovascular risk?]. AB - OBJECTIVE: The International Diabetes Federation (IDF) has recently published the new criteria for the diagnosis of metabolic syndrome. The aim of this study was to compare the clinical characteristics and cardiovascular risk of the new patients with MS compared to the previous National Cholesterol Education Program ATP III definition, its differential characteristics and cardiovascular risk. PATIENTS AND METHODS: Cross sectional study in a hypertension clinic. Coronary risk was calculated (Framingham function NCEP-ATP III) and other cardiovascular markers, urinary albumin excretion (UAE in mg/24 hours) and high sensitivity C reactive protein (CRP) were assessed. RESULTS: A total 2,404 patients were evaluated, 1,901 non-diabetic and 503 diabetic hypertensive subjects. The non diabetics 726 (38.2%) had MS with the previous NCEP ATP-III definition, the number increasing sharply to 1,091 (57.4%) with the new IDF definition. The proportion did not increase in diabetics (93% vs. 92%). Concordance in the diagnosis was 78% in non-diabetics and 91% in diabetics. The new patients had a similar coronary risk (Framingham) but lower values of other cardiovascular markers: logUAE 1.00 (0.49) mg/24 hours vs. 1.06 (0.55) mg/24 hours (p = 0.003), and CRP 1.9 (2.7) mg/L vs. 2.5 (3.2) mg/L (median, interquartile range; p < 0.001). CONCLUSIONS: The new IDF definition of MS increases the number of patients with MS. The new patients have a similar coronary risk (Framingham) but the new parameters used to assess cardiovascular risk (UAE and CRP) were lower. The relationship of the new definition of MS and cardiovascular risk remains to be defined. PMID- 17397566 TI - [Late-onset familial amyloid polyneuropathy in the Safor (Valencia) area: four case reports]. AB - Four cases of late onset familial amyloid polyneuropathy diagnosed in relatively advanced ages and within 3 families of our area where the disease had never been described are presented. The possible origin of the mutation and need to consider this diagnosis even when there is no known family background or when the age of presentation is late are commented. The nerve biopsy does not always show the deposits as these are patchy and DNA studies or detection of abnormal TTR in serum are necessary for the diagnosis. PMID- 17397567 TI - [Mixed hereditary polyposis with atypical extracolonic manifestations]. AB - We present the case of a 31 year-old-man with mixed hereditary polyposis and atypical extracolonic manifestations, as patent ductus arteriosus and mental retardation, with cranial hyperostosis. This is an extremely uncommon polyposis syndrome and has a moderate risk to progress to colon cancer. PMID- 17397568 TI - [The figure of Chief of Residents in the Residencing training: establishment and development in a Spanish hospital]. AB - Training qualified medical specialists is one of the great supports of the Spanish health care system. The teaching structure of the hospital is essential for the correct development of this training. The figure of the chief resident (CR) is widely developed in the USA and Canada, but not in our country. Including this figure in the teaching organigram of our hospital has meant one more advance in the search for improving our teaching capacity. In the following, we explain our experience during the introduction and later development of the figure of CR, stressing its theoretical functions, the notification mechanism, constitution of the Residents' Committee and later development of activities and tasks conducted. After three years, our experience is favorable, subject to modifications, but adaptable to the idiosyncrasy of each site. PMID- 17397569 TI - [Current role of statins in the stroke prevention]. AB - The role of lipids in stroke has been a traditionally debated subject. Many studies have recently demonstrated that reduction of cholesterol levels is associated to a lower stroke incidence. At present, statins play an essential role in both primary as well as secondary stroke prevention. More knowledge on the use of statins in stroke in order to be able to apply it in the daily clinical practice is needed. PMID- 17397570 TI - [Clostridium difficile associated diarrhea in the elderly patient]. AB - Clostridium difficile is the leading cause of infectious nosocomial diarrhea in the elderly. It may cause colitis of variable severity and extraintestinal involvement. Hand transmission is the most important, and it is related to prolonged antibiotic therapies in elderly patients with severe baseline disease. Diagnosis is based on culture and immunological tests to detect its toxins. When there is no response to conservative treatment (withdrawal of the antibiotic and support therapy), metronidazole or vancomycin should be given. Recurrences, up to 20%, are frequent. Preventive measures of its spreading are essential due to the elevated transmission through health care personnel and instruments. Detection of new, more virulent, gene variants of Clostridium difficile have increased the importance of this disease. PMID- 17397571 TI - [Acute abdominal pain with pathological chest X-ray]. PMID- 17397572 TI - [Chest pain and peripheral eosinophilia]. PMID- 17397573 TI - [Cutaneous necrosis in patient with chronic hepatitis]. PMID- 17397574 TI - [79 year old male with constitutional syndrome and hepatomegaly]. PMID- 17397575 TI - [Diagnostic and therapeutic approach to cryptogenic stroke: do we have all the information we need?]. PMID- 17397576 TI - [Metabolic syndrome and peripheral arterial disease]. PMID- 17397577 TI - [Analysis of the alternatives to transfusion in pre-surgical patients with low hemoglobins]. PMID- 17397578 TI - [Dysphagia in young patient with an atopic background]. PMID- 17397579 TI - [Cerebellar atrophy in a HIV/AIDS patient]. PMID- 17397580 TI - [Sudden disease and hemochromatosis]. PMID- 17397581 TI - [Ambulatory chest surgery]. PMID- 17397582 TI - [Resources and delays in the diagnosis of sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: The demand for consultations and diagnostic studies for sleep apnea hypopnea syndrome (SAHS) has increased, and this has led to considerable delays. We therefore need an updated evaluation of the diagnostic situation to serve as a management tool for specialists and health care administrations responsible for solving the problem. The objective of the present study was to carry out a descriptive analysis of the situation regarding the diagnosis of SAHS in Spanish hospitals. METHODS: We undertook a descriptive cross-sectional observational study. Public and private hospitals listed in the Ministry of Health's 2005 catalog of health care institutions were contacted, and those that routinely evaluate patients for SAHS were included in the study. The person in charge of each hospital filled in a questionnaire concerning the availability of resources and waiting periods for diagnosis. RESULTS: Of the 741 hospitals we contacted, 217 routinely evaluated patients for SAHS. In 88% of these, respiratory polygraphy (RP) (n=168) or polysomnography (PSG) (n=97) was available. The mean waiting period was 61 days for consultation and 224 days for RP. The mean number of RP systems was 0.99 per 100,000 inhabitants, while the recommended number is 3 per 100,000 inhabitants. The mean waiting period for PSG was 166 days. The mean number of PSG beds was 0.49 per 100,000 inhabitants, while the recommended number is 1 per 100,000. CONCLUSIONS: We observed a marked inadequacy of resources that has led to unacceptable waiting periods. While there has been a favorable change in the situation regarding SAHS diagnosis compared to previous studies, there is still room for improvement and it is urgent that healt hcare authorities allocate more resources to this public health problem. PMID- 17397583 TI - [Video-assisted thoracic surgery to treat spinal deformities: climbing the learning curve]. AB - OBJECTIVE: The aim of this study was to analyze the impact of the learning curve on the preliminary results of video-assisted thoracic surgery for spinal deformities in a general hospital setting. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 15 patients who underwent video assisted thoracic surgery performed by a multidisciplinary team comprising orthopedic and thoracic surgeons. Endoscopic anterior release and fusion were followed by posterior instrumentation in a single procedure. Demographic, orthopedic, morbidity, and mortality statistics were compiled for the 15 patients and compared to results reported for similar series. RESULTS: Endoscopic surgery was indicated for 15 patients: 11 women (73.3%) and 4 men (26.7%). The median age was 15 years (interquartile range [IQR], 14-19 years). Three patients (20%) required conversion to thoracotomy. There were 2 serious (13.3%) and 3 minor complications (20%). They all resolved satisfactorily and there was no perioperative mortality. The median Cobb angle was 71 degrees (IQR, 63.75 degrees -75.25 degrees ) before surgery and 41 degrees (IQR, 30 degrees -50 degrees ) after surgery. Median duration of surgery was 360 minutes (IQR, 300-360 minutes), duration of postoperative recovery unit stay was 1.5 days (IQR, 1-2.75 days), and total hospital stay was 11.5 days (IQR, 8.25-14 days). CONCLUSIONS: Despite the complexity of video-assisted thoracic surgical procedures, we believe they will become the standard approach to treating spinal deformities in the near future. By working together in general hospital settings, orthopedic and thoracic surgeons can help to overcome the steep yet manageable learning curve. PMID- 17397584 TI - [Validity and reliability of the St George's Respiratory Questionnaire in adults with cystic fibrosis]. AB - OBJECTIVE: To study self-perceived quality of life in adults with cystic fibrosis (CF), and to assess the validity of the St George's Respiratory Questionnaire (SGRQ) for use in these patients. PATIENTS AND METHODS: We studied 37 adults with CF who were in stable condition as indicated by their respiratory and nutritional status. Disease severity was assessed by spirometry in conjunction with a modified National Institutes of Health (NIH) scoring system and the Bhalla scale. Nutritional status was evaluated by measuring height and weight, calculating body mass index, analyzing bioelectric impedance, and performing various laboratory tests. The patients' quality of life was assessed using the SGRQ. RESULTS: SGRQ scores were higher (indicating poorer quality of life) among patients with CF than in the general population or among patients with chronic obstructive pulmonary disease. Internal consistency coefficients indicated the SGRQ had good reliability (Cronbach alpha 0.864). Women with CF tended to score higher than men, although this difference was not statistically significantly. A statistically significant relationship was observed between SGRQ score and severity of pulmonary impairment (the more severe the impairment, the worse the patient's quality of life). Statistically significant positive relationships were found between SGRQ domains and age, body mass index, and body fat percentage. Significant negative relationships were found between these domains and the modified NIH score, the Bhalla score, forced expiratory volume in 1 second (expressed as a percentage of predicted), and somatomedin C and zinc levels. CONCLUSIONS: Self-perceived quality of life is worse among adults with CF than in the general population or among patients with chronic obstructive pulmonary disease. The SGRQ is a valid instrument for analyzing health-related quality of life in adults with CF as it discriminates very well between different degrees of severity of pulmonary impairment and also have an appropriate intern consistency. PMID- 17397585 TI - [Spanish productivity in smoking research relative to world and European union productivity from 1999 through 2003, analyzed with the science citation index]. AB - OBJECTIVE: To analyze Spanish scientific productivity from 1999 through 2003 in the area of smoking research, in comparison with world and European Union research, based on data in the Science Citation (SCI) Index. MATERIAL AND METHODS: This bibliometric study was carried out by searching the title field of the Science Citation Index Expanded. Descriptive statistics with 95% confidence intervals (CI) were compiled. RESULTS: Two-hundred ninety-two documents on smoking by Spanish authors were located. The most productive subspecialties were the group comprised of public health, education and health economics with 57 articles, and areas of the respiratory system, experimental research, and internal medicine with 36 articles each. The journals that published the largest number of articles located were Medicina Clinica, with 35 articles (12%) and Archivos de Bronconeumologia with 20 (6.8%). International collaboration was undertaken with institutions in the United States of America and other European Union countries. The mean (SD) number of citations received was 5.12 (8.6) (range 59-0). Spain contributed 8.34% of the SCI-indexed smoking research overall and 12.85% of SCI-indexed smoking research from the European Union. CONCLUSIONS: Smoking research is developing appropriately in Spain and has high impact, even though output is modest. Factors associated with increased citation are international collaboration, language (being written in English), and having a non-Spanish first author. PMID- 17397586 TI - [Usefulness of endoscopic ultrasound-guided fine needle aspiration in the diagnosis of mediastinal lesions]. AB - OBJECTIVE: Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) is a safe and effective technique for the diagnosis of focal pancreatic lesions and enlarged abdominal lymph nodes. The aim of this study was to assess the usefulness of EUS-FNA in the diagnosis of mediastinal lesions. PATIENTS AND METHODS: A retrospective review was performed of all consecutive cases in which EUS-FNA was used for the diagnosis of a mediastinal lesion between January 2001 and September 2003. We used a radial echoendoscope to assess the characteristics of the lesion and a linear-array echoendoscope to perform transesophageal needle aspiration with a 22-gauge needle. Histopathology of the resected specimen was considered as the gold standard in surgically treated patients whereas cytology obtained by EUS-FNA was the gold standard when surgery was not indicated. RESULTS: EUS-FNA was performed in 59 patients with a total of 89 lesions with mean (SD) dimensions of 2.4 (2.0) cm x 1.6 (1.4) cm. Malignant lesions were larger than benign ones (short axis, 2.7 [1.4] as compared with 1.0 [0.9] cm; P< .001). The diagnosis was obtained for 53 patients (90%) and 81 lesions (91%) with a mean of 2 (1) passes per lesion. The sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy of EUS-FNA were 81%, 100%, 100%, 75%, and 88%, respectively, when analyzed by lesion, and 88%, 100%, 100%, 80%, and 92% when analyzed by patient. CONCLUSIONS: EUS-FNA is an effective technique for the diagnosis of mediastinal lesions. The likelihood of malignancy increases with size. PMID- 17397587 TI - [Innovative strategies to diagnose and monitor tuberculosis patients]. AB - Mycobacterium tuberculosis is the single most deadly microorganism worldwide. A third of the world population is thought to have latent tuberculosis and approximately 2 million people die of the disease each year. Short and closely supervised treatment regimens are needed, but it is also essential to develop new strategies to ensure prompt diagnosis of the disease. In particular, cheap methods are needed to tackle tuberculosis from a population perspective. The present article reviews the advances in immunology and molecular strategies for epidemiological diagnosis and monitoring of tuberculosis patients. PMID- 17397588 TI - [Advances in the study of infant lung function: forced expiratory maneuvers from an increased lung volume]. AB - Forced expiratory maneuvers from an increased lung volume in infants date from 1989 and consist of raising the inspiratory volume by applying a specific inflation pressure until a level close to the total lung capacity is reached. The chest and abdomen are then compressed by means of an inflatable jacket in order to obtain a forced expiratory flow-volume curve similar to that obtained for an adult. Forced expiration from an increased lung volume in infants is useful, just as the maneuver is in older patients, for studying airway function, diagnosing obstructive diseases early, and assessing response to treatment. The objective of this review is to provide information on the physiological bases and technical aspects of a lung function test that has proven highly useful for the study of the airways of healthy infants as well as those with respiratory diseases. PMID- 17397589 TI - [Occlusion of a pulmonary arteriovenous fistula with an amplatzer vascular plug]. AB - Pulmonary arteriovenous malformations are rare anomalies that carry a considerable risk of serious complications such as cerebral thromboembolism or abscess and pulmonary hemorrhage. The first-line treatment of such malformations is detachable coil or balloon embolotherapy. However, coils and balloons may migrate and cause paradoxical embolism especially in malformations with large arteriovenous shunts. We report a case in which we used a new vascular occlusion device (amplatzer vascular plug), to occlude a pulmonary arteriovenous fistula in a patient with Rendu-Osler-Weber syndrome. PMID- 17397590 TI - [Yellow nails syndrome as a form of presentation of pleural effusion]. PMID- 17397591 TI - [On the association between recurrent venous thromboembolic disease and hyperhomocysteinemia]. PMID- 17397592 TI - [Dermatofibrosarcoma protuberans]. AB - Dermatofibrosarcoma protuberans (DFSP) is a soft tissue neoplasm of intermediate malignancy that is initially localized to the skin from where it can invade deep structures (fat, fascia, muscle and bone). It is the most frequent fibrohistiocytic tumor, comprising approximately 1.8 % of all soft tissue sarcomas and 0.1 % of all cancers. It has an estimated incidence of 0.8-5 cases per one million persons per year. Treatment of localized disease consists in complete surgical excision of the lesion by conventional surgery with wide margins (>3 cm) or by micrographic Mohs surgery. Although the cases of metastatic DFSP do not reach 5 % of the total, almost all of them appear after previous local relapses. The prognosis for metastatic cases is very poor with a survival of less than 2 years following detection of metastatic disease. Patients with locally advanced DFSP are not candidates for an initial radical surgical therapy therefore neoadyuvant treatment is required prior to surgery in order to reduce tumor burden. In this regard, chemotherapy and radiotherapy have not been highly efficacious so it is necessary to consider new alternatives. The demonstration of the oncogenic power of the translocation COL1A1-PDGFB in DFSP has allowed the successful introduction of drug therapy with antagonists of the PDGFB receptor for metastatic or locally advanced cases. PMID- 17397593 TI - [Immunization against viral hepatitis: an obligatory recommendation in consults for sexually transmitted diseases]. AB - Numerous international institutions recommend vaccination against hepatitis B for all non-immune patients that consult for a sexually transmitted disease. It is a simple and safe procedure, frequently forgotten in dermatological clinics. Hepatitis B vaccine is administered via intramuscular route in the deltoid in 3 doses (at months 0, 1 and 6). The vaccine against hepatitis A is indicated for non-immune homosexual men that have a sexually transmitted disease. There exists a combined vaccine A + B that facilitates the administration in these cases. PMID- 17397594 TI - [Allergic contact dermatitis to temporary henna tattoos]. AB - INTRODUCTION: In the last years there have been increasing reports of adverse cutaneous reactions to temporary black henna tattoos. Black henna does not exist naturally, it is obtained from original henna after the addition of other compounds, among them paraphenylenediamine (PPD), that darken it and facilitate the process of tattoing. Paraphenylenediamine is an aromatic compound that presents cross reactions with other components that have a benzene ring in their molecular structure. Many of these products may be present in the daily life of any person. MATERIAL AND METHODS: We reviewed patients that have shown erythema, inflammation and/or vesiculation in a previously tattooed area. The patients have undergone a temporary tattoo in street stalls during the summer period (2004 2005). A total of five patients were included, there were four men and one woman with a mean age of 13 years (3-34) and a mean latency period of 9.4 days (5-14). Black ink and plastic stencils were used to perform the tattoo. None of the patients has had previous contact with hair dyes or tattoos and none of them referred a personal history of atopic dermatitis. Patch testing was carried out using the standard set of the Spanish Group for Research in Contact Dermatitis [GEIDC] (TRUE TEST, Pharmacia. Hillerod. Denmark), with readings at 48 and 96 hours. RESULTS: Sensitization to PPD is confirmed in three patients, one of them was also sensitized to formaldehyde. Hypopigmented scars persist in three patients. CONCLUSIONS: Black henna pseudotattoos are a source of sensitization to PPD with potential severe consequences in a medium to long term. Currently there is no specific legislation with respect to the practice of this type of tattoos in our country. PMID- 17397595 TI - [Results and assessment of photopatch testing in Spain: towards a new standard set of photoallergens]. AB - INTRODUCTION: While the standardization of exploration with photoallergy tests or photopatch testing runs its course in Europe, we have carried out an epidemiological study about the current situation of photoallergy in our country. MATERIAL AND METHODS: We have gathered the results of photopatch testing in seven hospital centres of Madrid, Cataluna, Galicia and Comunidad Valenciana during the years 2004 and 2005. The exploration has included, at least, the standard set of the Spanish Photobiology Group (GEF), with 16 (photo) allergens, that have been irradiated with 10 joules/cm(2). We have assessed the total number of explored patients, their sex, present, past or unknown relevance of positive photopatch testing, cross reactions, and allergens responsible for photosensitization. RESULTS: Of 224 patients explored by photopatch testing, 39.3 % show one or more positive tests. Seventy-one percent (103) were considered relevant with respect to clinical history, 14 cases (9.6 %) were cross reactions, and 28 (19.3 %) were considered of unknown relevance. The most prevalent allergens were nonsteroidal antiinflammatory drugs, specially ketoprophen (43 patients), followed by bencydamine (7 patients) and etofenamate (5 patients). The mixture of four sunscreens from the standard set of the GEF only detected 10 of 16 patients with photoallergy to sunscreens. Photopatch testing of unknown relevance was mainly due to antiseptics (fenticlor) and topical antihistamines. CONCLUSIONS: We propose the modification of the standard set of photoallergens from the GEF, that should include the majority of nonsteroidal antiinflammatory drugs and sunscreens available in Spain. Ketoprophen continues to be the most frequent photoallergen in our country. It is also important for the cross sensitizations that may present. Sunscreens should be explored separately and not in form of a mixture. PMID- 17397596 TI - [Neutrophilic dermatosis of the dorsal hands]. AB - Neutrophilic dermatosis of the dorsal hands is a recently described disorder. It is still debatable if it constitutes a separate entity, a variant of Sweet syndrome or pyoderma gangrenosum or an overlap disorder of both. We report the clinical features, histopathological findings, and evolution of a 35-year-old patient with the diagnosis of neutrophilic dermatosis of the dorsal hands and distant lesions on the forehead. PMID- 17397597 TI - [Fatal Sweet syndrome associated to chronic idiopathic systemic inflammatory response syndrome]. AB - Sweet syndrome is one of the cutaneous processes more frequently associated to systemic diseases. Its association to the systemic inflammatory response syndrome has rarely been described. We report a case of chronic and relapsing Sweet syndrome associated to a chronic and idiopathic systemic inflammatory response syndrome that lasted seven years and proved fatal to the patient. Among the rare cases of Sweet syndrome associated to a systemic inflammatory response syndrome that have been described there have not been any fatal cases as occurred with our patient. PMID- 17397598 TI - [Familial presentation of multiple scalp tumors]. AB - A 74-year-old woman was referred for evaluation and treatment of a tumor measuring 3 x 2 cm in the left preauricular region that has been progressively growing in the past years. Physical examination revealed an hemispheric and firm tumor with reddish coloration and telangiectases. The patient has been using a wig during the past 15 years to cover the scalp lesions. The patient underwent complete surgical excision of the scalp and the defect was reconstructed with partial-thickness grafts. Given the long-standing evolution and extension of the tumors involving the scalp and face we suspected a familial cylindromatosis syndrome therefore, we revised her six children observing scalp tumors in two daughters. The tumors were excised and the histological findings confirmed the diagnosis of cylindromas. PMID- 17397599 TI - [Incontinentia pigmenti: three new cases that demonstrate it is not only a matter of women]. AB - Incontinentia pigmenti is a rare, dominantly X-linked genodermatosis characterized by multisystemic involvement that is lethal prenatally in the majority of affected males and shows great clinical variability when it is expressed in women. Recently it has been shown that mutations of the gene NEMO/IKK-g located in Xq28 cause the expression of the disease, being only one mutation responsible for approximately 80 % of the cases. The diagnosis of incontinentia pigmenti is performed based on clinical features and family history with the support of histological findings. Nevertheless, as the gene responsible for the phenotype of the disease has been identified, a genetic study may be employed for doubtful cases. We report three cases of this entity (two women and one man) in different clinical stages of development that show the broad clinical spectrum we may encounter in the clinic. PMID- 17397600 TI - [Pachydermoperiostosis (Touraine-Solente-Gole syndrome). Case report]. AB - Pachydermoperiostosis or primary hypertrophic osteoarthropathy, also known as Touraine-Solente-Gole syndrome, is a rare process, frequently inherited. In its complete form it is characterized by pachydermia (thickening of the skin), skeletal changes (periostosis) and acropachia (digital clubbing). We report a patient that consulted for skeletal symptoms, as the acropachia and cutaneous manifestations (thickening of the skin of the face, scalp, hands and feet) went unnoticed due to their slow and progressive development. We review the characteristic features of this syndrome. We highlight the importance of ruling out secondary forms of hypertrophic osteoarthropathy and of a close follow-up of these patients because of complications that might develop on the long-term. PMID- 17397601 TI - [Giant dermatofibroma: case report and review of the literature]. AB - Dermatofibroma is a very frequent lesion that usually appears as a slowly growing nodule in the dermis, and preferentially involves the lower extremities of women. Several clinical variants have been described. Giant dermatofibroma has been defined as a rare variant of dermatofibroma measuring more than 5 cm that presents typical histological features and a benign biological behavior. We report the case of a 52-year-old man that presented a giant dermatofibroma with a diameter of 6 cm in the right shoulder and we review the few cases of this variant that have been described in the literature. PMID- 17397602 TI - [Subungueal verrucous and painful tumor]. PMID- 17397603 TI - [Intermittent vesiculous eruption in a child]. PMID- 17397604 TI - [Confidence intervals, "p"s and lampposts]. PMID- 17397605 TI - [Benign epithelioid fibrous histiocytoma]. PMID- 17397606 TI - [Eruptive pseudoangiomatosis]. PMID- 17397607 TI - [Burnout syndrome in Intensive Care Units]. PMID- 17397608 TI - [Care to the end-stage patient: help and obstacles perceived by Intensive Care nurses]. AB - BACKGROUND: The Intensive Care Unit (UCI) environment is not the most appropriate for the development of the end-of-life process, due to the fact that ICU is a hi tech setting and its focus is on curing and giving life support, rather than delivering palliative care to patients. AIMS: To investigate supportive behaviours and obstacles, and the nurses' demographic characteristics. METHOD: A descriptive correlational design was used in five tertiary Spanish hospitals. A convenience sample included 151 critical care nurses. A self-administered anonymous questionnaire (Beckstrand and Kirchhoff, 2005) was used to investigate supportive behaviours and obstacles perceived by nurses providing end-of-life care, in a scale from 0 to 5 (O = not help/obstacle; 5 = main help/obstacle). Some demographic data of the sample were also collected. FINDINGS: Nurses mean age was 35 (min. 22-max. 57; SD = 7,6) and had an average of 9,2 (min. 1-max. 30; SD = 6,9) years of experience working in ICU. Physicians agreeing on direction of patient care was perceived as the most supportive item (x = 4.46); whereas ethics committee constantly involved in the unit as the least supportive one (x = 2.93). The main obstacle for nurses was patient having pain that is difficult to control or alleviate (x = 4.38), and nurses knowing poor prognosis before family was seen as the less important obstacle (x = 1.37) Statistically significant correlations were found between nurses age and years of experience in ICU and their perception of some helps/obstacles. Statistically significant differences were found between nurses with postgraduate education in intensive care and those without it and their perception of some helps/obstacles. CONCLUSIONS: Intensive care nurses perceive adequate patients' pain management, agreement between health professionals on decision-making, and facilitating a comfortable environment for patients and families, during the whole end-of-life process as a priority. PMID- 17397609 TI - [Reliability of the measurement of glucose at the bedside of critical patients]. AB - INTRODUCTION: The objective of this study is to measure the reliability of three measurement methods at the bedside of the patient, of glucose in the critical patient compared with the measurement of glucose in the central laboratory. MATERIAL AND METHODS: Observational, perspective study developed in a polyvalent unit of 18 beds for four months. Patients who had arterial catheter were included. Eight samples obtained at the patient's bedside were compared with the plasma glucose (gold Standard): three in capillary blood, four in arterial blood and one in arterial blood gases from a syringe. The measurements at bedside were conducted with reactive strips MediSense Optium Plus and glucometer MediSense Optium. A comparison was made of the means used in the Student's T test and Bland and Altman analysis. RESULTS: We obtained 630 samples in 70 patients. Mean glucose (SD) in mg/dl was: a) capillary samples: 149 (38), 149 (35), 147 (37); b) arterial samples: 140 (34), 142 (35), 143 (35), 142 (34); arterial gas sample syringe: 143 (33); c) plasma glucose: 138(33). There were significant differences (p < 0.001) between plasma glucose and capillary samples but not with arterial samples (p=0.2). In the arterial samples, the presence of some factors, such as vasoactive drugs, glycated solution perfusion, insulin perfusion and plasma concentration of hemoglobin, increase error and dispersion regarding the gold standard. CONCLUSIONS: The measurement of glucose at bedside in critical patients is more reliable in arterial samples than in capillary ones. PMID- 17397610 TI - [Comparative study of maintenance of patency of triple lumen central venous catheter]. AB - OBJECTIVE: Establish an action protocol for the maintenance of patency of the three lumen central venous catheters (CVC) when any of them are not used. The main variable of interest was evaluate it 24 hours. MATERIAL AND METHOD: This is a blind, randomized clinical trial. Phase 1: Comparison of efficacy, for maintenance of patency of the lumen at 24 hours and on discharge of the patient, of sodium heparin 500 IU (control 1), versus sodium heparin 100 IU (study 1). Phase 2: Once efficacy of sodium heparin 100 IU (control 2) was confirmed, we compared it with saline solution (SS) (study 2). POPULATION: multiple diseased patients admitted to eight critical care unit who were carriers of 3 lumen CVC without previous coagulation disorders. ANALYSIS: quantitative variables with mean (SD) analyzed with Student's t test and qualitative ones with -2. ANALYSIS: SPSS 12.0 program. RESULTS: Phase 1: 128 catheters (control 1: 49, study 1: 79). Mean days of permanency with CVC: 5.4 (SD 6). At 24h, 100% of the CVC of both groups were patent. On discharge of the patient (n = 107) were not patent and 4.9% of the control group and 4.5% of the study group 1. Phase 2: 95 catheters (control 2: 38, study 2: 57) Mean days of permanency: 4.87 (SD 5). At 24h (n = 95), at 72 h (n = 66) and on discharge (n = 43), 100% of the CVC of both groups were patent. No significant difference was established in the two study phases between the groups studied with days of permanence, laboratory values and anticoagulant treatment. CONCLUSIONS: SS is as effective as 100 or 500 IU of heparin for maintenance of patency of CVC. PMID- 17397611 TI - [Farewell]. PMID- 17397612 TI - [Open letter to the members of SERAM]. PMID- 17397613 TI - [Case 1. Classic Pelizaeus-Merzbacher Disease]. PMID- 17397614 TI - [Hysterosalpingography: how, when, what for?]. AB - Histerosalpingography is the most commonly used technique in the evaluation of infertility. It has traditionally been considered the gold standard for assessment of Fallopian tubes giving information about their patency and morphology. It is also recommended for the study of the uterine cavity. With a good knowledge about the anatomy and the variants of the normality, and a good technique, we can do a good differential diagnosis between normality and pathology, which probably it will need more tests. PMID- 17397615 TI - [Clinical practice guidelines (I): elaboration, implementation and evaluation]. AB - Clinical practice guidelines are defined as a set of systematically developed recommendations that aim to guide the decision-making process. In the last ten years, interest in clinical practice guidelines has grown steadily as they have enabled a point of encounter between research and clinical practice. Unlike classical protocols, usually based on consensus or opinion and a narrative review of the available scientific literature, clinical practice guidelines are developed by multidisciplinary teams, review the evidence exhaustively and systematically, evaluate the quality of the information, and propose specific recommendations in line with the quality and design of the studies available. The later stages of dissemination, implementation, and posterior evaluation of their impact complete the cycle for dynamic change in clinical practice. PMID- 17397616 TI - [Clinical practice guidelines (II): searching and critical evaluation]. AB - Clinical practice guidelines have unique characteristics of the Internet era in which they are starting to be increasingly popular. The fact that they are often elaborated by governmental agencies and are not published in conventional journals means that they may not be accessible using the usual search methods employed for other types of scientific studies and documents (clinical trials, reviews, etc.). The Internet has become an essential tool for locating clinical practice guidelines, and meta-search engines, specific databases, directories, and elaborating institutions are of special importance. The relative lack of indexing of clinical practice guides means that Medline and Embase are not as useful in this context as in searching for original studies. With the aim of evaluating the validity, reproducibility, and reliability of clinical practice guidelines, a series of European institutions designed a tool to evaluate clinical practice guidelines at the end of the 1990s. This instrument, named AGREE, aims to offer a framework for the evaluation of the quality of clinical practice guidelines. It can also be useful in the design of new clinical practice guidelines as well as in the evaluation of the validity of guidelines to be updated or adapted. The AGREE instrument has become the reference for those that use guidelines, those that elaborate them, and for healthcare providers. PMID- 17397617 TI - [Case 2. Diffuse hepatic angiosarcoma secondary to thorotrast deposit]. PMID- 17397618 TI - [Integrated PET/CT scanner in oncology applications: a radiologic perspective]. AB - OBJECTIVE: To describe our experience with an integrated PET/CT system in the clinical applications in Oncology. MATERIAL AND METHODS: An integrated PET/CT scanner is an in-line system combining a full-ring detector PET and a multidetector row helical CT in one machine. This is a multidisciplinary technique involving a nuclear medicine physician, a radiologist, a radiopharmacologist, and a physicist, with distinct tasks and working together. The clinical indications for PET scans are determined by the National Health System, which in Madrid is coordinated by Agencia Lain Entralgo, and are performed by a nuclear medicine physician with the help of a radiologist for CT interpretation. The combined PET/CT scans are supervised and interpreted by both a nuclear medicine physician and a radiologist. RESULTS: Between September 2003 and August 2005, a total of 2459 examinations were performed at our institution: 2200 were PET scans with low-dose noncontrast-CT for attenuation correction (clinical indications approved by NHS), and 259 were combined PET/CT scans with full-diagnostic enhanced-CT (in the setting of research programmes). The overall distribution of clinical indications for the 2459 examinations were: lung cancer 14.7%, solitary lung nodule characterization 7%, lymphoma 23.5%, colon cancer 18.5%, gastric cancer 1.9%, brain tumors 2.6%, head-neck tumors 5.5%, thyroid cancer 5%, breast cancer 4%, unknown origin cancer 4.3%, epilepsy 1.4%, others 11.6%. CONCLUSION: In our experience, the integrated PET/CT system has advantages over the stand-alone PET and/or CT. However, more studies, as the ongoing research programmes at our facility are needed for the diagnostic validation of this technique. PMID- 17397619 TI - [Mammographic findings in microcalcifications associated with breast cancer after neoadjuvant chemotherapy]. AB - OBJECTIVE: To evaluate the mammographic findings in microcalcifications associated with breast cancer after neoadjuvant chemotherapy. MATERIAL AND METHODS: From January 2000 to May 2005, a total of 99 breast cancer patients underwent neoadjuvant chemotherapy. Ten patients had microcalcifications on mammograms prior to treatment. We evaluated the evolution of the tumor and of the microcalcifications, correlating the imaging findings with the clinical and histological manifestations. RESULTS: Four different patterns of evolution were observed for the microcalcifications: the number of particles increased in two cases, remained stable in three, decreased in four, and in one patient the microcalcifications disappeared. The size of the tumor decreased in all cases. CONCLUSIONS: After neoadjuvant chemotherapy microcalcifications can evolve unpredictably and independently of tumor response. PMID- 17397620 TI - [Case 3. Hydrocarbon-induced pneumonitis]. PMID- 17397621 TI - [Severe hydroureteronephrosis associated to asymptomatic giant anterior sacral meningocele: a case report and review of the literature]. AB - We present the case of a woman with Marfan's syndrome presenting with a clinical picture of acute gastroenteritis in whom severe bilateral hydroureteronephrosis associated to a neurogenic bladder and a giant anterior sacral meningocele was diagnosed incidentally. The importance of this case lies in the fact that the patient was asymptomatic despite the significant visceral repercussions already occurring that led to questioning of whether MRI follow-up would still be advisable even in the absence of symptoms. PMID- 17397622 TI - [Spontaneous spinal subdural hematoma: MRI findings in two cases]. AB - Spontaneous spinal subdural hematoma is an uncommon finding. This report of two cases of spinal subdural hematoma (one hyperacute and one subacute) discusses the current theory of how it develops, the MRI findings, its differentiation from spinal epidural hematoma, and the value of MRI in monitoring patients undergoing conservative treatment. The most significant MRI findings in the hyperacute phase were isointense signal on T1-weighted sequences and hyperintense signal on T2 weighted sequences. Moreover, T2-weighted gradient-echo sequences showed a hypointense peripheral ring. In the subacute phase, a hyperintense signal was seen on T1-, T1-fat-suppression, and T2-weighted sequences. PMID- 17397623 TI - [Heterotopic mesenteric ossification]. AB - We report a case of heterotopic bone formation in the mesentery. A 64-year-old man presented with abdominal pain and stiffness two months after undergoing several surgical interventions for hemorrhagic colitis. The diagnosis was heterotopic mesenteric ossification. We describe the imaging findings and review the literature. PMID- 17397624 TI - [Myofibroblastic inflammatory tumor of the lung]. AB - Myofibroblastic inflammatory tumor is a controversial entity that shows great variability in clinical presentation, histological findings, evolution, and prognosis. It is a rare cause of primary lung tumor in adults; however, it is the most common cause of lung tumors in children. The diagnosis is fundamentally histological, although histological diagnosis is not easy because myofibroblastic inflammatory tumor is characterized by a polymorphic cellular infiltration of variable cellular composition that could be similar to other diseases such as lymphoma or low-grade sarcoma. We report the case of a 23-year-old woman in whom a solitary pulmonary nodule was discovered incidentally at plain-film chest x ray. PMID- 17397628 TI - [Letter to the President of Sociedad Espanola de Radiologia Vascular e Intervencionista]. PMID- 17397629 TI - [Smoking, obesity and body fat distribution are independently associated with insulin resistance and other cardiovascular risk factors]. AB - BACKGROUND AND AIM: In recent years, smoking has been shown to increase insulin resistance. However, other studies have failed to demonstrate this association after correcting for confounding factors such as age, gender, body mass index (BMI) or waist to hip ratio (WHR). This study was conducted to elucidate the relative contributions of smoking, obesity and body fat distribution on insulin resistance and other cardiovascular risk factors. PATIENTS AND METHODS: Cases and controls matched for age, gender and degree of obesity. Evaluations included anthropometrical and biochemical assessments with body mass index (BMI), waist to hip ratio (WHR), lipid profiles and insulin resistance determined by HOMA (Homeostasis Model Assessment). RESULTS AND CONCLUSION: A total of 126 patients (52 smokers and 74 non-smokers) participated in the study. Of all the patients, 22 (17.5%) were lean, 49 (38.9%) overweight and 55 (43.7%) obese. Multivariate stepwise linear regression showed an association of WHR (beta = 0.414, p < 0.001), BMI (beta = 0.211, p = 0.012), the number of smoked cigarettes per day (beta = 0.200, p = 0.011) and serum triglycerides levels (beta = 0.241, p = 0.007) on insulin resistance (R = 0.628, F = 13.841, p < 0.001). An independent effect of smoking on triglycerides levels was also shown. Therefore, smoking, obesity and body fat distribution are independently associated with insulin resistance and lipid profile. PMID- 17397630 TI - [Effect of a global intervention in the integral control of multiple risk factors in patients at high or very high cardiovascular risk. CIFARC 2 project]. AB - OBJECTIVES: To analyze the effect of a global approach on patients with high cardiovascular risk for the integral control of cardiovascular risk factors. PATIENTS AND METHODS: Multicentric prospective study of patients with high vascular risk, followed-up for one year in internal medicine consultations. We measured the grade of control over major cardiovascular risk factors (hypertension, dyslipidemia, diabetes mellitus, tobacco, obesity) before and after the one-year follow-up period. RESULTS: We studied 456 patients (56% males; age: 66.1, standard deviation: 11.5 years); 54.4% were diabetics, 58.0% had target organ damage and 46.2% had cardiovascular disease. Blood pressure control varied from 50.5 to 60.6; LDL-cholesterol control from 44.7 to 58.1%; diabetes control from 56.5 to 56.6%; tobacco control from 63.0 to 87.5% and obesity control from 60.0 to 55.4%. Integral control of all cardiovascular risk factors varied from 10.7 to 23.2% of patients. Factors independently associated to lack of integral control were: diabetes (Odds Ratio [OR]: 4.42; 95% confidence interval [95%CI]: 2.75-7.14), basal systolic blood pressure (OR: 1.03; 95%CI 1.02 1.05) and basal body mass index (OR: 1.08; 95%CI: 1.02-1.14). Lack of integral control was independently associated to the incidence of cardiovascular events (OR: 2.00; 95%CI: 1.09-5.35). CONCLUSIONS: A global approach on the patient with high cardiovascular risk duplicated the integral control of the five main risk factors. Diabetes and obesity were factors which made integral control difficult. Lack of integral control increased two times the risk of cardiovascular events. PMID- 17397631 TI - [Nonagenarian's risk profile for living in a nursing home. The NonaSantfeliu study]. AB - OBJECTIVE: By means of a cross-sectional study, describe the differential characteristics between a group of inhabitants of a same municipality over 89 years of age who are permanently institutionalized and those of the same age who continue to live in the community, trying to identify a profile of nonagenarians with high risk of living in a nursing home. MATERIAL AND METHODS: One hundred and eighty six nonagenarians were studied: 137 (74%) were living in their home and 49 (26%) in a nursing home. Sociodemographic data, ability to perform basic daily activities with Barthel Index (BI) or instrumental activities with Lawton and Brody Index (LI), cognition with the Spanish version of the Mini-Mental State Examination (MEC) and comorbidity (Charlson Index) were evaluated. Cardiovascular risk factors and other prevalent diseases were also studied. RESULTS: A total of 143 women (76.5%) and 43 men with a mean age of 93.06 years were studied. A predominance of women, single persons, worse IL, and greater consumption of drugs was observed in the bivariate analysis in the nonagenarians living in residences and widowers than in those living in the community. The multivariate analysis showed that the features of being single and having a lower IL were independent factors of living in a nursing home. CONCLUSIONS: The main difference between nonagenargian patients who live in the community and those in residences is that the latter are mainly single and have a low score on a scale that quantities instrumental activities of the daily life. PMID- 17397632 TI - [Effect of renin-angiotensin system in Fabry disease associated proteinuria]. AB - AIM: To describe the prevalence of renal involvement in Fabry disease patients and determine the role of ACE inhibitors in its treatment. MATERIAL AND METHODS: We studied a family of eight members, diagnosed of Fabry disease after demonstrating alpha-galactosidase deficiency and genetic mutation. Serial biochemical analyses were performed every six months during the three years of follow-up: creatinine, urea, creatinine clearance, proteinuria, microalbuminuria, urinary sediment, blood pressure and glycemia. If urinary alterations were detected, ACE inhibitors were started. At the end of the study, a simple and Doppler ultrasonography was performed. RESULTS: Six of eight patients presented microalbuminuria during the follow-up. Only one of these patients did not develop proteinuria. ACE inhibitors therapy decreased proteinuria in all six patients, however, this decrease was not complete in two of them: in one proteinuria was detected and in the other microalbuminuria persisted. Kidney involvement was not dependent on enzymatic substitution therapy. Renal ultrasonography was normal in patients without biochemical sign of renal affection. In one patient with proteinuria at the moment of the ultrasonography, slightly increased resistance indexes were detected. CONCLUSIONS: Renal involvement is very frequent in patients with Fabry disease (in six of eight in our series). ACE inhibitors are effective in controlling proteinuria in patients with microalbuminuria y proteinuria. These data must be confirmed in larger series. Doppler ultrasonography fails in early renal involvement detection, but as it constitutes an easy and not dangerous technique, it should be done routinely in Fabry patients in order to evaluate its role in the follow-up of these patients. PMID- 17397633 TI - [Diarrhea in patients with percutaneous endoscopic gastrostomy probe]. PMID- 17397634 TI - [Multiple furunculosis after a trip to Africa]. PMID- 17397635 TI - [Symptoms in the era of technology]. PMID- 17397636 TI - [37 Year old woman with abdominal discomfort and splenic focal lesion]. PMID- 17397637 TI - [Are benzodiazepines correctly used in the elderly?]. AB - Benzodiazepines are substances susceptible of producing drug dependence in addition to tolerability to sedative effects and frequent interactions when associated with other drugs. Thus, use of long half life benzodiazepines and inadequate intermediate and short half life dosage is related with falls and fractures in elderly patients. A retrospective study was conducted on the official prescriptions made during 2000-2003 and the daily dose defined per thousand patients/day (DDD/I) was used as a measurement of drug consumption. Prescription of benzodiazepines that are eliminated by oxidative metabolism should be revaluated and modified by other of conjunctive metabolism when treating elderly patients to decrease adverse reactions such as excess sedation, risk of falls and associated fractures. PMID- 17397638 TI - [Vegetarian diets; effect on health]. AB - Vegetarian diets are those diets mainly based on the consumption of vegetable product, but that also permit consumption of eggs and milk. The American Dietetic Association made a declaration on these vegetarian diets in which they stated that <>. Some studies have shown beneficial results in obesity, cancer, Parkinson disease, hypertension, type 2 diabetes mellitus and urinary stones, compared with the omnivorous. The possible theoretical benefits in some diseases has been seen in the medical practice (diabetes mellitus, obesity, cardiovascular risk). However more studies are needed in the case of Parkinson's disease and rheumatoid arthritis. PMID- 17397639 TI - [Intermediate Care Units dependent on Internal Medicine in a hospital without an Intensive Care Unit]. AB - Intermediate care units are considered a solution for care of patients who require superior care than those in a normal hospitalization ward but are not amenable to intensive therapy. We present our experience with an open model, with a responsible person of the ward appointed to the Internal Medicine Department, that could be assumed in other hospitals without Intensive Care Unit in which these units would also avoid unnecessary transfers or make it possible to optimize them so that these are done under better conditions. PMID- 17397640 TI - [Assessment of terminal disease in Internal Medicine]. PMID- 17397641 TI - [Elevated levels of IgE and lymphomatoid papulosis (subtype B) as background of well-differentiated lymphocyte B cell lymphoma]. PMID- 17397642 TI - [Influence of age in creatinine clearance]. PMID- 17397643 TI - [Treatment with pegylated interferon and ribavirin in patients co-infected by human immunodeficiency virus and hepatitis C virus]. PMID- 17397644 TI - [Pneumocystis, the controversy continues]. PMID- 17397645 TI - Surface plasmon resonance biosensor for enrofloxacin based on deoxyribonucleic acid. AB - A DNA-based surface plasmon resonance biosensor for enrofloxacin was developed. Heating denatured DNA immobilized on the gold-coated glass surface was exploited. The immobilization was performed by a layer-by-layer co-deposition with a cationic polymer. The sensor performance was tested with real biological probes. Direct and simple determination of enrofloxacin in milk samples was demonstrated. The sensor response obeys Langmuir binding isotherm being almost linear until about 20 microg mL(-1). The detection limit in milk samples was estimated to be 3 microg mL(-1). PMID- 17397646 TI - Polypyrrole-polyvinyl sulphonate film based disposable nucleic acid biosensor. AB - Double stranded calf thymus deoxyribonucleic acid entrapped polypyrrole-polyvinyl sulphonate (dsCT-DNA-PPy-PVS) films fabricated onto indium-tin-oxide (ITO) coated glass plates have been used to detect organophosphates such as chlorpyrifos and malathion. These disposable dsCT-DNA-PPy-PVS/ITO bioelectrodes have been characterized using cyclic voltammetry, Fourier-transform-infra-red (FTIR) spectroscopy and atomic force microscopy (AFM), respectively. These biosensing electrodes have a response time of 30s, are stable for about 5 months when stored in desiccated conditions at 25 degrees C and can be used to amperometrically detect chlorpyrifos (0.0016-0.025 ppm) and malathion (0.17-5.0), respectively. The additive effect of these pesticides on the amperometric response of the disposable dsCT-DNA-PPy-PVS/ITO bioelectrodes has also been investigated. PMID- 17397647 TI - One-shot screen-printed thylakoid membrane-based biosensor for the detection of photosynthetic inhibitors in discrete samples. AB - Screen-printing technology offers the possibility to produce a large number of sensors at low cost. Thus, due to their intrinsic characteristics and reproducibility, screen-printed electrodes can be used in the development of disposable electrochemical devices. In the present work, carbon-based screen printed electrodes (SPCEs) have been used to develop a one-shot-measure biosensor for the detection of photosynthetic inhibitors in discrete samples. The measurement was based on the electrochemical evaluation of the activity of photosystem II (PSII), a protein complex present in photosynthetic organisms and involved in the photosynthesis. The biosensor was prepared by the modification of the working electrode of a SPCE, using thylakoid membranes extracted from spinach leaves. The modified sensors were then used as one-shot system to measure the presence of PSII activity inhibitors in discrete standard solutions. The coupling of the developed biosensor with a custom-made cell made it possible to perform tests using only 50 microL of total sample volume with a measurement time of 10 min. Inhibition curves were recorded for some photosynthetic inhibitors in a concentration range of 10(-6) to 10(-8) molL(-1). A reproducibility (relative standard deviation, R.S.D.%) of 10% was found and the calculated limit of detections (LODs) were in the nanomolar range. The effect of storage on sensitivity and reproducibility of a biosensor prepared by direct lyophilisation of thylakoid membranes on the electrode surface was also evaluated, confirming the possible use of the modified sensor up to one week after the preparation. Measurements on real samples were also reported, comparing the results with those obtained using a fluorescence-based commercial instrument for the analysis of photosynthetic inhibitors. PMID- 17397648 TI - A novel flow injection potentiometric graphite coated ion-selective electrode for the low level determination of uranyl ion. AB - Solution studies on the binding properties of uranyl ion toward four different recently synthesized benzo-substituted macrocyclic diamides L1-L4 revealed the occurrence of a 1:1 complexation of the ligands with UO(2)(2+) ion, with a stability order of L2>L1>L4>L3. Consequently, L2 was used as a suitable neutral ionophore for the preparation of novel polymeric membrane (PME) and coated graphite (CGE) UO(2)(2+)-selective electrodes. The electrodes exhibit a Nernstian behavior for UO(2)(2+) ions over wide concentration ranges (1.0 x 10(-6) - 1.0 x 10(-1) M for PME and 1.0 x 10(-7) - 1.0 x 10(-1) M for CGE) and very low limits of detection (8.0 x 10(-7) M for PME and 7.3 x 10(-8) M for CGE). The proposed potentiometric sensors manifest advantages of fast response and, most importantly, good selectivity with respect to many alkali, alkaline earth, transition, and heavy metal ions. The potentiometric responses of the electrodes are independent of the pH of the test solution in the pH range 2.9-3.7. The CGE was used in flow injection potentiometry and resulted in well defined peaks for uranyl ions with stable baseline, excellent reproducibility and very high sampling rate of 170 injections per hour. The proposed FIP system was used for the determination of trace uranyl ions in real and synthetic samples. PMID- 17397649 TI - PVC membrane electrode based on triheptyl dodecyl ammonium iodide for the selective determination of molybdate(VI). AB - Novel polyvinyl chloride (PVC) membrane electrodes based on triheptyl dodecyl ammonium iodide have been developed. In the presence of 12.5 mM H(2)O(2), these electrodes are capable of determining molybdate(VI) ion. The electrodes exhibit near-Nernstian responses over a wide concentration range (2.0 x 10(-6) to 5.0 x 10(-3) M). The proposed electrodes demonstrate satisfying selectivity for molybdate(VI) ion in the presence of a wide variety of anions other than iodide, and can be used in the pH range 5.0-7.0. Moreover, the electrodes show an average response time of 2-3 min and can be used over a period of 2 months without any significant deviation being observed. In the light of our results, the response mechanism of the electrode is discussed and HMoO(2)(O(2))(2)(-) is suggested as the response ion. The proposed electrode has been used to measure molybdenum in ore samples, and the results were in agreement with those obtained by means of ICP analysis. PMID- 17397650 TI - Dependence on the electric power of the immersion-angle dependence of the resonant-frequency shift of a quartz crystal microbalance in a liquid. AB - We have investigated the immersion-angle dependence of the series resonant frequency shift, DeltaF, of the quartz crystal microbalance, QCM, in a Newtonian liquid from the point of view of the supplied electric power level. In the low electric power levels, the immersion-angle dependence and the transition phenomenon of the DeltaF are observed. However, when the higher electric power levels are supplied to the QCM, the region of the transition phenomenon of the DeltaF decreases rapidly with an increase in the electric power level and disappears above 1.5 dBm. That is, above 1.5 dBm, the DeltaF values have only the values of 90 degrees immersion angle in all immersion angles. We suggest that the electric power is very important factor for the DeltaF in a liquid. PMID- 17397651 TI - Preparation for nitrocellulose membrane-poly (vinyl alcohol)-ionic imprinting and its application to determine trace copper by room temperature phosphorimetry. AB - Nitrocellulose membrane-poly (vinyl alcohol)-ionic imprinting (NCM-PVA-I-I) was prepared using Cu2+ as template. The cavity in NCM-PVA-I-I matched Cu2+ very well and the selectivity was high. Cu2+ entered the cavity and then could form ionic association ([Cu2+] x [(Fin-)2]) with the anion of fluorescein (Fin-) outside the cavity by electrostatic effect. [Cu2+] x [(Fin-)2] could emit strong and stable room temperature phosphorescence on NCM-PVA-I-I. Its DeltaI(p) was proportional to the content of Cu2+. Based on the above facts, a new method for the determination of trace copper by solid substrate-room temperature phosphorimetry (NCM-PVA-I-I-SS-RTP, SS-RTP is the abbreviation of solid substrate-room temperature phosphorimetry) using NCM-PVA-I-I technique has been established. The linear range of this method was 2.00-144.00 fg Cu2+ spot(-1) (sample volume: 0.40 microL spot(-1), corresponding concentration: 5.00-360.00 pg mL(-1)), and the detection limit calculated by 3Sb/k was 0.43 fg Cu2+ spot(-1) (corresponding concentration: 1.1 x 10(-12) g mL(-1), n=11). Samples containing 2.00 and 144.00 fg Cu2+spot(-1) were measured, respectively, for seven times and R.S.D.s were 3.5% and 4.7%. NCM-PVA-I-I-SS-RTP could combine very well the characteristics of both the high sensitivity of SS-RTP and the high match and selectivity of NCM-PVA I-I, and it was rapid, accurate, sensitive and with good repeatability. It has been successfully applied to determine trace copper in human hair and tea samples. PMID- 17397652 TI - Direct screening of tetracyclines in water and bovine milk using room temperature phosphorescence detection. AB - A fast and simple flow-through optosensor was designed and characterized for the direct screening of four tetracycline (TCC) antibiotics (tetracycline, oxytetracycline, chlortetracycline and doxycycline) in water and bovine milk samples. The proposed optosensor provides rapid binary yes/no overall responses, being appropriate for the screening of this family of antibiotics above or below a pre-set concentration threshold. The experimental set-up is based on a flow injection manifold coupled on-line to a phosphorescence detector. Aliquots of the samples are pretreated with Eu(III) to form room temperature phosphorescent metal chelates and injected in the flow manifold. Those chelates are then on-line retained on a conventional flow-cell (packed with polymeric Amberlite XAD-4 particles) which is placed inside the cell holder of the phosphorimeter. After the emission is registered, the antibiotic-metal complexes are eluted from the packed resin with 1M HCl (for milk samples a second regeneration step, using methanol, should be performed). A sample throughput of about 20 samples per hour was obtained. Optimum experimental conditions include a pH 9, a Eu(III) concentration of 2 x 10(-4) M and 8 mM sodium sulphite as chemical deoxygenant. The phosphorescence emitted by the europium-TCC complexes was measured at 394 and 617 nm for excitation and emission wavelengths, respectively. The unreliability region, given by the probability of false positives and false negatives, respectively (set at 5% in both cases) was in the range between 0.2 and 11.6 nM for detection of tetracyclines in water samples (at a cut-off level of 4 nM) and in the range between 165 and 238 nM for detection of tetracyclines in milk (cut off level fixed at the normative EU level of 200 nM). Finally, the applicability of the proposed screening optosensor was tested for the reliable control of tetracyclines in contaminated and uncontaminated water and milk samples. PMID- 17397653 TI - On-line selective detection of antioxidants free-radical scavenging activity based on Co(II)/EDTA-induced luminol chemiluminescence by flow injection analysis. AB - This study establishes a new method to analyze the radical scavenging activity of antioxidants based on the luminol-H(2)O(2)-Co(II)/EDTA chemiluminescence and flow injection analysis. The method is based on the catalytic oxidation of hydrogen peroxide by Co(II)/EDTA complex, forming a free radical flux that can produce a stable chemiluminescence signal which is attenuated in the presence of antioxidants. A properly designed FIA manifold and the appropriate regulation of the chemiluminescence-reagent mixture enabled the establishment of a reaction sensitive analytical procedure that minimizes oxidant-antioxidant interactions while favors the inhibition effect of antioxidants on the free radicals flux. In that manner, the uncontrolled experimental variability induced by side-reactions occurring antagonistically is reduced. The method was examined in-vitro for the continuous monitoring of the generation of oxygen-derived free radicals and antioxidants, which is closer to in-vivo conditions, with three common antioxidants (ascorbic acid, glutathione and uric acid). All three antioxidants were found to inhibit the luminescent signal with strict logarithmic linear mode, yielding calibration curves rectilinear in the range of 5 x 10(-8) - 5 x 10(-5) molL(-1) and detection limits at the 10(-8) molL(-1) levels. The F-statistic was employed to assess the ability of the method to detect differences in the activity of the examined antioxidants. The results suggest that the proposed method can be used efficiently for the detection of free radical activity in real samples. PMID- 17397654 TI - Analysis of antioxidants using a capillary electrophoresis with chemiluminescence detection system. AB - We developed a capillary electrophoresis with chemiluminescence detection system using 2-methyl-6-p-methoxyphenylethynylimidazopyrazinone as a chemiluminescence reagent for determination of antioxidants of superoxide anions. 2-Methyl-6-p methoxyphenylethynylimidazopyrazinone reacted with superoxide anions generated through the reaction of hypoxanthine and xanthine oxidase, and then emitted chemiluminescence. Suppression of the chemiluminescence in the presence of antioxidants for superoxide anions was introduced as a detection principle for antioxidants into the capillary electrophoresis with chemiluminescence detection system. After optimizing the analytical conditions, various antioxidants, such as superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin, were subjected to the present system. They gave negative peaks due to the quenching effect; the detection limits of superoxide dismutase, nitroblue tetrazolium, ascorbic acid, and catechin were 1, 100, 100, and 10 microM, respectively (S/N=2). A model sample consisting of superoxide dismutase and nitroblue tetrazolium was satisfactorily separated and detected within ca. 10 min. We also applied the present system to analysis of catechin in green tea as a real sample. PMID- 17397655 TI - Influence of a stacking phenomenon on the results of Hadamard transform capillary electrophoresis. AB - Advantageous features (enhanced sensitivity and low baseline noise) of the multiplex measuring procedure, the Hadamard transform capillary electrophoresis (HT-CE), are experimentally demonstrated and critically discussed. To perform a perfect multiplex experiment in CE, sample species need to be dissolved in the background electrolyte medium and have very low concentration. The mismatch of electric conductivity resulting from a sample dissolved in water or in a separation buffer diluted with water will lead to sample stacking and corrupting the anticipated outcome. The multiplex measurements were carried out with benzyltriethylammonium bromide, resorcinol and 3,5-dihydroxybenzoic acid in the phosphate buffer, 511 sample/buffer injections were performed into the capillary according to the pseudorandom binary sequence. The averaged electropherogram of the single injection was calculated from the detection signal with the aid of the Fourier transform. The results illustrate the detrimental effect of sample matrix dilution with water and the effect of increased initial sample concentration on the multiplex measurement in CE. Multiplex advantage, in theory possible in the HT-CE, can be obtained at low concentration levels feasible with laser induced fluorescence and optically gated sampling. To achieve successful multiplex measurements with the UV detector, the single injection signal should be approximately at the baseline noise level. PMID- 17397656 TI - Preparation of an ion-exchangeable polymer bead wrapped with bilayer membrane structures for high performance liquid chromatography. AB - We synthesized a chromatographic packing material that has a non-covalently attached dihexadecyl phosphate (DHP) bilayer membrane structure on a CA08S, a nonporous-type cationic polymer bead with a diameter ranging from 11 to 14 microm. Confocal fluorescence microscopic and differential scanning calorimetric analyses of the DHP-CA08S complex revealed that the DHP bilayer membrane structures were formed on the surface of the CA08S polymer beads. When the functionality of the DHP-CA08S complex was evaluated in the ion-exchange HPLC of proteins, the retention behavior of the proteins on the DHP-CA08S complex column totally mirrored the anionic property of the DHP bilayer membrane surface, not the cationic property of the CA08S bead. Methylene blue (MB) was eluted from the DHP-CA08S complex column in the isocratic elution mode, but not at all from a CK08S column, a styrene-divinylbenzene based cation-exchange polymer. When the column temperature was elevated from 50 to 60 degrees C, the peak shape of MB on the DHP-CA08S complex column became fairly sharp without a change in its peak area, which mirrored the characteristic phase transition of the DHP bilayer membrane formed on the DHP-CA08S complex. PMID- 17397657 TI - Simple and rapid methods for the fabrication of polymeric and glass chips for using in analytical chemistry. AB - This paper describes simple and rapid methods for the fabrication of glass and polymeric chips for routine analytical applications. The methods are easily interfaced to the general laboratory environment and do not require special clean room facilities or expensive instruments. Glass microchips were fabricated by etching with HF solution. Microfluidic channels were designed with CAD program and transferred onto a sheet of commercial polymeric self-adhesive (PSA) film by a cutter plotter. The PSA film was used as a mask for etching process. The etching rate was about 7 microm min(-1). A cover glass plate was sealed on the top of etched substrate by using polycellulose (cellophane). Polymeric microchips were fabricated by sawing with a jigsaw. Commercial polycarbonate (PC) was used as a substrate and two iron sheets were used as leader masks. While this restricts us to the fabrication of straight channels, it is however, much faster and less complicated than the other methods. The chip comprised three polymeric plates and the channels were created in the middle plate. Thermal bonding was used to bond three layers of the microfluidic chip. With this method, we could achieve simple channels with the width of about 200 microm. The channel depth depends on the polymeric plate thickness. Fabricated channels were accurate without any sinuosity or sideshow. PMID- 17397658 TI - Combining information from headspace mass spectrometry and visible spectroscopy in the classification of the Ligurian olive oils. AB - An electronic nose and an UV-Vis spectrophotometer, in combination with multivariate analysis, have been used to verify the geographical origin of extra virgin olive oils. Forty-six oil samples from three different areas of Liguria were included in this analysis. Initially, the data obtained from the two instruments were analysed separately. Then, the potential of the synergy between these two technologies for testing food authenticity and quality was investigated. Application of Linear Discriminant Analysis, after feature selection, was sufficient to differentiate the three geographical denominations of Liguria ("Riviera dei Fiori", "Riviera del Ponente Savonese" and "Riviera di Levante"), obtaining 100% success in classification and close to 100% in prediction. The models built using SIMCA as a class-modelling tool, were not so effective, but confirmed that the results improve using the synergy between different analytical techniques. This paper shows that objective instrumental data related to two important organoleptic features such as oil colour and aroma, supply complementary information. PMID- 17397659 TI - Characterization and discrimination of different aged 'Aceto Balsamico Tradizionale di Modena' products by head space mass spectrometry and chemometrics. AB - This work can be seen as an attempt to develop an analytical procedure in the context of quality control and authenticity assessment of typical food. To this aim, head-space mass spectrometry (HS-MS) coupled with multivariate data analysis, is proposed as a fast technique for furnishing a clear visualization and a suitable interpretation of the ageing process of 'Aceto Balsamico Tradizionale di Modena' (ABTM) and, for classifying products of different age. Considering the complexity of this food matrix, due to its traditional making procedure, the obtained instrumental data have first been analysed by parallel factor analysis (PARAFAC), an extension of principal component analysis to higher order arrays, in order to visualise the 'natural' grouping of vinegar samples and to inspect producers similarity/dissimilarity. On the basis of the PARAFAC results a reasonable class partition with respect to ageing was accomplished and both linear discriminant analysis (LDA) and soft independent modelling of class analogy (SIMCA) were applied as classification tools. Furthermore, it has been shown that discrimination on age basis can be improved by using feature selection in the wavelet domain through WPTER algorithm. PMID- 17397660 TI - Precise measurement of Fe isotopes in marine samples by multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS). AB - A novel analytical technique for isotopic analysis of dissolved and particulate iron (Fe) from various marine environments is presented in this paper. It combines coprecipitation of dissolved Fe (DFe) samples with Mg(OH)(2), and acid digestion of particulate Fe (PFe) samples with double pass chromatographic separation. Isotopic data were obtained using a Nu Plasma MC-ICP-MS in dry plasma mode, applying a combination of standard-sample bracketing and external normalization by Cu doping. Argon interferences were determined prior to each analysis and automatically subtracted during analysis. Sample size can be varied between 200 and 600 ng of Fe per measurement and total procedural blanks are better than 10 ng of Fe. Typical external precision of replicate analyses (1S.D.) is +/-0.07 per thousand on delta(56)Fe and +/-0.09 per thousand on delta(57)Fe while typical internal precision of a measurement (1S.E.) is +/-0.03 per thousand on delta(56)Fe and +/-0.04 per thousand on delta(57)Fe. Accuracy and precision were assured by the analysis of reference material IRMM-014, an in-house pure Fe standard, an in-house rock standard, as well as by inter-laboratory comparison using a hematite standard from ETH (Zurich). The lowest amount of Fe (200 ng) at which a reliable isotopic measurement could still be performed corresponds to a DFe or PFe concentration of approximately 2 nmol L(-1) for a 2 L sample size. To show the versatility of the method, results are presented from contrasting environments characterized by a wide range of Fe concentrations as well as varying salt content: the Scheldt estuary, the North Sea, and Antarctic pack ice. The range of DFe and PFe concentrations encountered in this investigation falls between 2 and 2000 nmol L(-1) Fe. The distinct isotopic compositions detected in these environments cover the whole range reported in previous studies of natural Fe isotopic fractionation in the marine environment, i.e. delta(56)Fe varies between -3.5 per thousand and +1.5 per thousand. The largest fractionations were observed in environments characterized by redox changes and/or strong Fe cycling. This demonstrates the potential use of Fe isotopes as a tool to trace marine biogeochemical processes involving Fe. PMID- 17397661 TI - Volatiles emission patterns of different plant organs and pollen of Citrus limon. AB - The volatiles emitted in vivo by different plant parts of Citrus limon (Rutaceae) have been identified by mean of head space-solid phase micro extraction (HS-SPME) and gas chromatography coupled with mass spectrometry (GC-MS) analyses. In particular, the profiles of flower buds, mature flowers, petals, stamens, gynaecium, pericarp of unripe and ripe fruits, young and adult leaves and pollen have been examined. Furthermore, the essential oil obtained from expression of ripe pericarp was studied. Volatiles were produced in distinctive amounts by the different plant organs, creating an interesting contrast, particularly within the flower parts: the highest amount of limonene (62.5%) was emitted by gynaecium, followed by stamens (22.9%) and petals (3.1%). Pollen did not produce limonene at all. The same compound is contained in higher amounts in the young leaves than in old ones (65.3% versus 30.1%). A possible defensive role of limonene and other volatiles, mainly terpene aldehydes, produced by young leaves has been hypothesized. PMID- 17397662 TI - Microwave-assisted solvent extraction of solid matrices and subsequent detection of pharmaceuticals and personal care products (PPCPs) using gas chromatography mass spectrometry. AB - Concentrations of pharmaceuticals and personal care products (PPCPs) in natural solids remain largely unknown. Contributing to this, is a lack of methods permitting the simultaneous detection of the diverse, low-level contaminants present in these complex matrices. We have developed a microwave-assisted solvent extraction (MASE)-based method targeting seven diverse PPCPs (caffeine, 17beta estradiol, ibuprofen, ketoprofen, musk ketone, naproxen, and triclosan) and a molecular marker for fecal waste (epicoprostanol). The method consisted of optimizing the following variables: derivatization of the polar target analytes, silica gel open column clean-up, and gas chromatographic-mass spectrometric (GC MS) analysis of sample extracts for analysis and detection of the compounds noted above. Testing of the method on spiked soil allowed for 89.6+/-2.89% recovery of three target compounds and 25.0+/-1.93% recovery of five of the compounds. Although the latter recoveries were low, the precision across all recoveries was high, suggesting good reproducibility in application of the method. Furthermore, we suspect that matrix effects are likely responsible for the lower recoveries. Techniques with the exclusive incorporation of organic solvents were found inapplicable in the study of a pharmaceutical salt, diphenhydramine HCl. Application of the developed method to sediment collected directly downstream of the effluent pipe of a wastewater treatment plant allowed detection of ibuprofen, naproxen, ketoprofen, and epicoprostanol at ng-mug per gram dry weight concentrations. The observation of acidic pharmaceuticals, previously believed to exhibit insignificant sorption to solid matrices, in the tested sediment samples, coupled with application of biosolids for agricultural purposes, demonstrates the need for expanded investigation of PPCP contamination of natural solid matrices. PMID- 17397663 TI - Determination of Henry's law constants for low volatile mixed halogenated anisoles using solid-phase microextraction. AB - Trihalogenated anisoles (THAs) that have been identified at low concentration levels (ngL(-1)) in drinking water are suspected of causing odor episodes, which are a frequent source of complaint by consumers. Henry's law constant (K(H)) is an important parameter in controlling the diffusion of organic compounds from the water to the vapor-phase, so its evaluation is of significance in the study of odor events. In this paper, the K(H) of a wide range of trihalogenated anisoles- in its dimensionless form [Formula: see text]--were calculated at two temperatures, 45 and 22 degrees C using equilibration partitioning in a closed system and headspace microextraction (EPICS-SPME). Two methodological approaches, Ramachandran and Dewulf, were used for the assessment of the Henry's law constant. Nevertheless, to apply these methods to THAs, a relatively narrow headspace/water volume ratio range (80/1-8/1) is required. At these conditions, a linearity (r(2)) using Ramachandran's theoretical relationship from 0.9276 to 0.9989 was obtained and the variability (R.S.D.%) when Dewulf's theoretical relationship was employed was lower than 20% (n=5). PMID- 17397664 TI - Spectrophotometric determination of aqueous cyanide using a revised phenolphthalin method. AB - This paper focuses on a revision of the phenolphthalin method for cyanide analysis, with the intent of producing a robust and sensitive spectrophotometric method. Limitations of the phenolphthalin chemistry were overcome by the addition of EDTA to the reagent. The revised reagent was found to have suitably fast kinetics, a linear dynamic range of 0.01-3.0 ppm cyanide, and a detection limit of 5 ppb. The method was tested for interferences and applied to the determination of cyanide in environmental waters. PMID- 17397665 TI - Development of a competitive immunoassay for efavirenz: hapten design and validation studies. AB - The reverse transcriptase inhibitor efavirenz (EFV) is widely used in human immunodeficiency virus (HIV) therapy. Knowledge of the plasma and intracellular concentrations of the drug is of prime importance to get further insight into EFV action in vivo and would be useful for therapeutic drug monitoring (TDM). The aim of this study was to develop a sensitive and specific competitive enzyme immunoassay (EIA) for EFV in biological fluids. Two haptens that differed by the position of the linker were synthesized using two different ways and coupled to BSA. Anti-EFV polyclonal antibodies (pAb) were raised in rabbits using the corresponding immunogens. By comparing results obtained with EIA study with those observed with high-performance liquid chromatography (HPLC) we have shown that the position of the linker appears to be crucial for the specificity of the pAb. EIA was then developed in microtitration plates using the most specific pAb. The assay was performed on a minimum of 30 microL of plasma. It showed good precision and efficiency as well as good cross-validation with HPLC. The lowest limit of quantification (LLOQ) was 150 pg mL(-1), i.e., a value at least 10 times lower than those currently achieved using previously described techniques. This EIA should be useful in the clinical laboratory for monitoring patients during antiretroviral therapy especially young children as well as for measuring EFV in intracellular studies requiring lower amounts of biological material. PMID- 17397666 TI - Prediction of ozone tropospheric degradation rate constants by projection pursuit regression. AB - Quantitative structure-property relationship (QSPR) models were developed to predict degradation rate constants of ozone tropospheric and to study the degradation reactivity mechanism of 116 diverse compounds. DUPLEX algorithm was utilized to design the training and test sets. Seven molecular descriptors selected by the heuristic method (HM) were used as inputs to perform multiple linear regression (MLR), support vector machine (SVM) and projection pursuit regression (PPR) studies. The PPR model performs best both in the fitness and in the prediction capacity. For the test set, it gave a predictive correlation coefficient (R) of 0.955, root mean square error (RMSE) of 1.041 and absolute average relative deviation (AARD, %) of 4.663, respectively. The results proved that PPR is a useful tool that can be used to solve the nonlinear problems in QSPR. In addition, methods used in this paper are simple, practical and effective for chemists to predict the ozone degradation rate constants of compounds in troposphere. PMID- 17397667 TI - The use of prazosin for the treatment of trauma nightmares and sleep disturbance in combat veterans with post-traumatic stress disorder. PMID- 17397668 TI - Abstracts of the 62nd Annual Meeting of the Society of Biological Psychiatry. May 17-19, 2007. San Diego, California, USA. PMID- 17397669 TI - Arterial ultrasonography and tonometry as adjuncts to cardiovascular risk stratification. AB - Myocardial infarction and stroke often occur without prior warning in asymptomatic individuals. Identifying individuals at risk is important for cost effective use of preventive therapies. Algorithms based on risk factors statistically associated with cardiovascular events classify individuals into high-risk, intermediate-risk, or low-risk categories. However, more than one third of adults in the U.S. are in the intermediate-risk category, and decisions regarding therapy are challenging in this subset. Testing for alterations in arterial function and structure that predate cardiovascular events may help refine cardiovascular risk assessment in the intermediate-risk group and identify candidates for aggressive therapy. Vascular ultrasonography and tonometry are promising test modalities for assessment of arterial function and structure in asymptomatic subjects. Several prospective studies have shown that measures of arterial function and structure provide prognostic information incremental to conventional risk factors. Standardization of methodology and establishment of quality control standards in the performance of these tests could facilitate their integration into clinical practice as adjuncts to existing cardiovascular risk stratification algorithms. PMID- 17397670 TI - Inverse relationship of blood pressure levels to sudden cardiac mortality and benefit of the implantable cardioverter-defibrillator in patients with ischemic left ventricular dysfunction. AB - OBJECTIVES: This study was designed to evaluate the relationship among blood pressure (BP) levels, risk of sudden cardiac death (SCD), and benefit of the implantable cardioverter-defibrillator (ICD) in patients with ischemic left ventricular (LV) dysfunction. BACKGROUND: Low BP has been shown to be associated with increased mortality in patients with LV dysfunction and heart failure. We hypothesized that increasing BP levels are associated with a reduction in the risk of SCD in this population, thereby limiting ICD efficacy in a lower-risk subset. METHODS: The independent contribution of systolic blood pressure (SBP) and diastolic blood pressure (DBP) to outcome was analyzed in 1,231 patients enrolled in the prospective MADIT-II (Multicenter Automatic Defibrillator Implantation Trial II). RESULTS: Multivariate analysis showed that in the conventional therapy arm of the trial, 10-mm Hg increments in systolic BP were independently associated with a respective 14% (p = 0.01) and 16% (p = 0.04) reduction in the risk of cardiac mortality and SCD; similar trends were shown for DBP. Defibrillator therapy provided the least survival benefit to patients in the lower-risk, upper SBP (>130 mm Hg) and DBP (>/=80 mm Hg) quartiles (hazard ratio 1.04 [p = 0.89] and 1.05 [p = 0.88], respectively), whereas a respective 39% and 38% (p = 0.002) reduction in the risk of death with ICD therapy was observed among patients with lower BP values. CONCLUSIONS: In patients with ischemic LV dysfunction, SBP and DBP levels show an inverse correlation with sudden cardiac mortality. These noninvasive hemodynamic parameters may be useful for identifying lower-risk patients, in whom the benefit of primary defibrillator implantation is more limited. PMID- 17397671 TI - The elusive scourge of sudden cardiac death: is rational decision making possible? Should there be standards of risks and predictions in medicine? PMID- 17397672 TI - Prevalence and severity of left atrial edema detected by electron beam tomography early after pulmonary vein ablation. AB - OBJECTIVES: The aim of this study was to investigate the prevalence and severity of left atrial (LA) edema after pulmonary vein (PV) ablation and its effect on the cardiac function. BACKGROUND: Though extensive LA catheter ablation has been demonstrated to be more effective in curing paroxysmal atrial fibrillation (PAF) than segmental ostial pulmonary vein isolation (S-PVI), it might cause life threatening complications, including congestive heart failure associated with LA edema. METHODS: Fifty patients underwent S-PVI (Group S) and 27 underwent circumferential PV antrum ablation (Group C) for drug-refractory PAF. Enhanced electron beam tomography (EBT) was performed before, 1 or 2 days after, and 1 month after the PV ablation, and transthoracic ultrasound cardiography (UCG) was performed 1 month after the PV ablation in all patients. RESULTS: The EBT assessment revealed LA edema immediately after the PV ablation in 47 Group S patients and all Group C patients. The severity of the LA edema, number of radiofrequency applications, and amount of radiofrequency energy delivered during the PV ablation was significantly greater in Group C than in Group S. One month after the PV ablation, in all patients, the EBT assessment revealed that those edematous changes had disappeared, and the UCG assessment showed no reduction in the cardiac function. CONCLUSIONS: Left atrial edema was observed in a large portion of the patients immediately after the PV ablation, and the severity of the LA edema depended on the extent and amount of the radiofrequency energy delivered in the PV ablation. The LA edema soon disappeared naturally and did not reduce the cardiac function. PMID- 17397673 TI - Isolated atrial segment pacing: an alternative to His bundle pacing after atrioventricular junctional ablation. AB - OBJECTIVES: This study was designed to investigate a practical alternative to His bundle pacing after atrioventricular (AV) junctional ablation by pacing a small area of isolated atrial tissue surrounding the AV node. BACKGROUND: His bundle pacing is preferred after AV junctional ablation in patients with refractory atrial fibrillation. However, it is technically difficult and not clinically useful at the present time. METHODS: This study was conducted in an isolated working swine heart model (n = 5), with real-time imaging capabilities. A small area of atrial tissue surrounding the AV node and the His bundle was isolated using sequential radiofrequency ablation lesions. RESULTS: Complete AV block created by segmental atrial isolation was achieved in 5 of 5 experiments. The isolated atrial segment was bordered by the ablation lines, the tricuspid annulus, and the AV node-His bundle. The AV conduction was characterized using a pacing electrode implanted into the isolated atrial segment. Pacing from the atria, the ventricles, and the isolated atrial segment at different rates confirmed complete bidirectional block between the atria and isolated area, whereas antegrade and retrograde AV nodal conduction between the isolated atrial segment and the ventricles remained intact. Pacing from the isolated area produced minimal changes in systolic left ventricular pressure compared with baseline sinus rhythm (mean -2 mm Hg). CONCLUSIONS: Isolation of a small area of atrial tissue surrounding the AV node is feasible by transcatheter radiofrequency ablation. This procedure may be a useful alternative to conventional AV junctional ablation because it can create complete AV block, while in effect permitting the equivalent of His bundle pacing after AV junctional ablation. PMID- 17397674 TI - Associations of gender and etiology with outcomes in heart failure with systolic dysfunction: a pooled analysis of 5 randomized control trials. AB - OBJECTIVES: This study sought to explore the gender-related differences in etiology and outcomes in chronic heart failure (HF) patients from 5 randomized trials. BACKGROUND: Each year, 550,000 new cases of HF are identified; however, there remain limited data on gender-related differences in etiology and outcomes among patients with HF with systolic dysfunction. METHODS: We analyzed data from 8,791 men and 2,851 women randomized in 5 clinical trials (PRAISE [Prospective Randomized Amlodipine Survival Evaluation], PRAISE-2, MERIT-HF [Metoprolol Extended Release Randomized Intervention Trial in Heart Failure], VEST [Vesnarinone Trial], and PROMISE [Prospective Randomized Milrinone Survival Evaluation]) to explore gender-related differences in etiology (ischemic vs. nonischemic) and outcomes (all-cause mortality and death or all-cause hospitalization). Hazard ratios (HR), 95% confidence intervals (CIs), and Kaplan Meier survival curves were generated by gender and etiology. RESULTS: A total of 18% of ischemic and 31% of nonischemic patients were women. Irrespective of etiology, women were older, more ethnically diverse, and had higher systolic blood pressures, more diabetes, and severe HF symptoms, but less often smoked or had prior myocardial infarctions than men. Mean ejection fractions were similar between women (23.6%) and men (23.2%). The 1-year Kaplan-Meier survival estimates varied by gender and etiology (female nonischemics, HR 0.88 [95% CI 0.85 to 0.89]; female ischemics, HR 0.83 [95% CI 0.81 to 0.85]; male nonischemics, HR 0.84 [95% CI 0.83 to 0.85]; male ischemics, HR 0.79 [95% CI 0.78 to 0.81]). After adjustment, female gender (HR 0.77 [95% CI 0.69 to 0.85]) and nonischemic etiology (HR 0.80 [95% CI 0.72 to 0.89]) were associated with longer survival time. Time to death or hospitalization was longer among nonischemics (HR 0.83 [95% CI 0.78 to 0.89], p < 0.0001); however, female gender was not significantly associated with the composite outcome (HR 1.01 [95% CI 0.95 to 1.08]). CONCLUSIONS: Our data clarify that outcomes differ by both gender and etiology among patients with HF with systolic dysfunction. Understanding these differences may lead to better management of HF patients and improved overall prognosis. PMID- 17397675 TI - Telomere length of circulating leukocytes is decreased in patients with chronic heart failure. AB - OBJECTIVES: This study sought to test the hypothesis that patients with chronic heart failure (CHF) have shorter telomeres compared with age-balanced and gender balanced healthy individuals. BACKGROUND: Telomere length is considered to be a marker of biological aging. Chronic heart failure might be viewed as a condition associated with accelerated biological aging. METHODS: The telomere length ratio of leukocytes was determined prospectively by a quantitative polymerase chain reaction-based method in a case-control setting involving 803 participants: 183 healthy individuals and 620 CHF patients, ages 40 to 80 years, New York Heart Association functional class II to IV, and left ventricular ejection fraction of 0.40 or less. RESULTS: The median telomere length ratio was 0.64 (interquartile range [IQR] 0.47 to 0.88) in CHF patients compared with 1.05 (IQR 0.86 to 1.29) in control patients (p < 0.001). The telomere length ratio in CHF patients related to severity of disease (median value [IQR] of patients with New York Heart Association class II, III, or IV function was 0.67 [0.48 to 0.92], 0.63 [0.46 to 0.86], and 0.55 [0.46 to 0.75], respectively; p for trend <0.05). In addition, telomeres were shorter in patients with an ischemic compared with a nonischemic etiology of CHF. Patients with none, 1 (coronary, cerebral, or peripheral vascular disease), 2 (any combination of the previous), or 3 atherosclerotic manifestations had a median (IQR) telomere length of 0.72 (0.51 to 1.01), 0.65 (0.48 to 0.87), 0.48 (0.39 to 0.72), and 0.43 (0.27 to 0.67), respectively (p for trend <0.001). CONCLUSIONS: Telomere length is shorter in patients with CHF compared with age-balanced and gender-balanced control patients, and related to the severity of disease. In addition, telomere length was incrementally shorter according to the presence and extent of atherosclerotic disease manifestations. PMID- 17397676 TI - Improved outcomes after aortic valve surgery for chronic aortic regurgitation with severe left ventricular dysfunction. AB - OBJECTIVES: Among patients undergoing aortic valve surgery for chronic aortic regurgitation (AR), we sought to: 1) compare survival among those with and without severe left ventricular dysfunction (LVD); 2) identify risk factors for death, including LVD and date of operation; and 3) estimate contemporary risk for cardiomyopathic patients. BACKGROUND: Patients with chronic AR and severe LVD have been considered high risk for aortic valve surgery, with limited prognosis. Transplantation is considered for some. METHODS: From 1972 to 1999, 724 patients underwent surgery for chronic AR; 88 (12%) had severe LVD. They were propensity matched to patients with nonsevere LVD to compare hospital mortality, interaction of operative date with severity of LVD, and late survival. Propensity score adjusted multivariable analysis was performed for all 724 patients to identify risk factors for death. RESULTS: Survival was lower (p = 0.04) among patients with severe LVD than among matched patients with nonsevere LVD (30-day, 1-, 5-, and 25-year survival estimates were 91% vs. 96%, 81% vs. 92%, 68% vs. 81%, and 5% vs. 12%, respectively). However, survival of patients with severe LVD improved dramatically across the study time frame (p = 0.0004): hospital mortality decreased from 50% in 1975 to 0% after 1985, and time-related survival in patients with severe LVD operated on since 1985 became equivalent to that of matched patients with nonsevere LVD (p = 0.96). CONCLUSIONS: Neutralizing risk of severe LVD has improved early and late survival such that aortic valve surgery for chronic AR and cardiomyopathy is no longer a high-risk procedure for which transplantation is the best option. PMID- 17397677 TI - Improved surgical outcome for chronic severe aortic regurgitation with severely depressed left ventricular systolic function. PMID- 17397678 TI - Measurement of collagen and smooth muscle cell content in atherosclerotic plaques using polarization-sensitive optical coherence tomography. AB - OBJECTIVES: The purpose of this study was to investigate the measurement of collagen and smooth muscle cell (SMC) content in atherosclerotic plaques using polarization-sensitive optical coherence tomography (PSOCT). BACKGROUND: A method capable of evaluating plaque collagen content and SMC density can provide a measure of the mechanical fidelity of the fibrous cap and can enable the identification of high-risk lesions. Optical coherence tomography has been demonstrated to provide cross-sectional images of tissue microstructure with a resolution of 10 mum. A recently developed technique, PSOCT measures birefringence, a material property that is elevated in tissues such as collagen and SMCs. METHODS: We acquired PSOCT images of 87 aortic plaques obtained from 20 human cadavers. Spatially averaged PSOCT birefringence, Phi, was measured and compared with plaque collagen and SMC content, quantified morphometrically by picrosirius red and smooth muscle actin staining at the corresponding locations. RESULTS: There was a high positive correlation between PSOCT measurements of Phi and total collagen content in all plaques (r = 0.67, p < 0.001) and in fibrous caps of necrotic core fibroatheromas (r = 0.68, p < 0.001). Polarization sensitive optical coherence tomography measurements of Phi demonstrated a strong positive correlation with thick collagen fiber content (r = 0.76, p < 0.001) and SMC density (r = 0.74, p < 0.01). CONCLUSIONS: Our results demonstrate that PSOCT enables the measurement of birefringence in plaques and in fibrous caps of necrotic core fibroatheromas. Given its potential to evaluate collagen content, collagen fiber thickness, and SMC density, we anticipate that PSOCT will significantly improve our ability to evaluate plaque stability in patients. PMID- 17397679 TI - Angiotensin receptor-1 blocker inhibits atherosclerotic changes and endothelial disruption of the aortic valve in hypercholesterolemic rabbits. AB - OBJECTIVES: We sought to examine the effect of angiotensin receptor blocker (ARB) on the formation of lesions in the aortic valves of hypercholesterolemic rabbits. BACKGROUND: Recently, atherosclerosis has been recognized as a mechanism that is responsible for calcific aortic stenosis. The effect of ARBs might help to prevent aortic stenosis because they have multiple antiatherosclerotic effects. METHODS: Male Japanese white rabbits (n = 36) were separated as follows: control with chow diet (C) and vehicle (V) groups, both of which were fed a 1% cholesterol diet for 8 weeks, and an ARB group (A), which was fed a 1% cholesterol diet for 8 weeks with ARB (olmesartan, 1 mg/kg/day) for the last 4 weeks. RESULTS: This dose of olmesartan did not affect either blood pressure or cholesterol levels. Dietary cholesterol induced fatty deposition with macrophage accumulation and osteopontin coexpression in valve leaflets, whereas ARB decreased macrophage accumulation (% area: V, 9.3 +/- 0.34; A, 1.4 +/- 0.30; p = 0.003) and osteopontin expression (p = 0.017). Angiotensin-converting enzyme was also up-regulated in V and decreased by olmesartan (p = 0.015). Immunohistochemistry with anti-CD31 antibody revealed that dietary cholesterol disrupted and olmesartan preserved endothelial integrity on the lesion-prone aortic side of the valve (% CD31-positive circumference: V, 30 +/- 3.7; A, 62 +/- 4.8; p = 0.003). Numbers of alpha-smooth muscle actin-positive myofibroblasts were increased in V and decreased by olmesartan (p = 0.003). Real-time polymerase chain reaction revealed that increased amounts of messenger ribonucleic acid for osteoblast-specific transcription factor core binding factor alpha-1 in V were diminished by olmesartan. CONCLUSIONS: Atherosclerotic changes in the aortic valves of rabbits fed with cholesterol were inhibited by ARB, whereas endo thelial integrity was preserved and transdifferentiation into myofibroblasts and/or osteoblasts in valve leaflets was inhibited. PMID- 17397680 TI - Initial worldwide experience with the WATCHMAN left atrial appendage system for stroke prevention in atrial fibrillation. AB - OBJECTIVES: This study assessed the feasibility of implanting a device in the left atrial appendage (LAA) in patients with atrial fibrillation (AF) to prevent thromboembolic stroke. BACKGROUND: Meta-analyses confirmed that in cases of left atrial thrombus in nonrheumatic AF patients approximately 90% of them are in the LAA. METHODS: The WATCHMAN Left Atrial Appendage System (Atritech Inc., Plymouth, Minnesota) is a nitinol device implanted percutaneously to seal the LAA. Patients were followed by clinical and transesophageal echocardiography at 45 days and 6 months with annual clinical follow-up thereafter. RESULTS: Sixty-six patients underwent device implantation. Mean follow-up was 740 +/- 341 days. At 45 days, 93% (54 of 58) devices showed successful sealing of LAA according to protocol. Two patients experienced device embolization, both successfully retrieved percutaneously. No embolizations occurred in 53 patients enrolled after modification of fixation barbs. There were 2 cardiac tamponades, 1 air embolism, and 1 delivery wire fracture (first generation) with surgical explantation but no long-term sequelae for the patient. Four patients developed a flat thrombus layer on the device at 6 months that resolved with additional anticoagulation. Two patients experienced transient ischemic attack, 1 without visible thrombus. There were 2 deaths, neither device related. Autopsy documented a stable, fully endothelialized device 9 months after implantation. No strokes occurred during follow-up despite >90% of patients with discontinuation of anticoagulation. CONCLUSIONS: Preliminary data suggest LAA occlusion with the WATCHMAN System to be safe and feasible. A randomized study is ongoing comparing oral anticoagulation with percutaneous closure. PMID- 17397681 TI - Watching the watchman. PMID- 17397682 TI - President's page: Achieving the heights of quality with electronic health records. PMID- 17397683 TI - A call for development of comprehensive therapy for dyslipidemia. PMID- 17397685 TI - Depression and heart failure: why the link continues to elude us. PMID- 17397687 TI - Mental health and work. PMID- 17397688 TI - Stress, coping, and alcohol expectancies in rural African-American women. AB - African-American women report less alcohol and other drug (AOD) use compared with Caucasian women. However, health care professionals cannot afford to dismiss AOD use as a significant problem for some African-American women. Although we know much more about AOD problems in women in general, we still lack information on AOD disorders and associated factors among rural African-American women. This article focuses on examining rural African-American women (n=142) with and without AOD disorders on the following variables: stressors, uplifts, coping, and alcohol expectancies. PMID- 17397689 TI - Transitional discharge based on therapeutic relationships: state of the art. AB - The Transitional Discharge Model (TDM) has been used to facilitate effective discharge from psychiatric hospital to community. A summary of the research to date on TDM is given. The model is based on the provision of therapeutic relationships to provide a safety net throughout the discharge and community reintegration processes. These relationships include both staff and peer involvement; hospital inpatient staff continue to remain involved with the client until a therapeutic relationship is established with a community care provider, and peer support is offered from a former consumer of mental health services who is currently living in the community. Studies found that the TDM facilitates increased discharge rates, lower readmission rates, and cost savings--making it a collaborative, cost-effective method of providing quality patient care and positive health care outcomes. The TDM Plan of Research is presented and reflects the collaboration between stakeholders and the integration of consumer advocacy, clinical practice, education, policy, and research in various countries. PMID- 17397690 TI - Medical/psychiatric comanagement by nurse practitioners in chronic hepatitis C treatment: a case study. AB - This article aims to increase access to care for patients with chronic hepatitis C who have medical and psychiatric comorbidities by reporting on a clinical advanced practice nursing comanagement model for evaluation, treatment, and follow-up care. PMID- 17397691 TI - Postnatal depression: prevalence, mothers' perspectives, and treatments. AB - This article presents an overview of research on postnatal depression conducted with postnatal women only. Research spanning three areas is reviewed, namely, prevalence, mothers' experiences ,and treatments. The review shows prevalence rates varying from 4.4% to 73.7%, with the most recent systematic review suggesting a rate of 13%, indicating a serious clinical issue for nurses providing postnatal care to mothers. Mothers' experiences of living with postnatal depression exemplify such feelings as loneliness, anxiety, hopelessness, and loss of control at a time when expectations of joyousness are anticipated. Results of research on treatment options show limited success with antidepressant medications, some success with psychotherapeutic options, and the importance of social support for mothers with postnatal depression. Comparisons of studies are made from methodological perspectives, drawing on their strengths and limitations. Overall, this review highlights the high prevalence rates of postnatal depression, negative feelings exemplified by mothers' living with this condition, and the limited success of treatment options available to mothers. Postnatal depression is a significant clinical issue for nurses providing postnatal care for mothers and is underassessed, misunderstood, and very often poorly treated. PMID- 17397692 TI - Integrating an evidenced-based research intervention in the discharge of mental health clients. AB - The purpose of this Canadian qualitative study was to explore the experiences of clinical staff who implemented a research intervention: the Transitional Discharge Model (TDM). The TDM provided mental health clients who were hospitalized with peer support and an inpatient staff member to bridge the therapeutic transitioning from hospital staff to the community care provider. Staff from three tertiary care mental health facilities in Canada identified their learning needs in regard to providing the intervention. An educational program was developed and delivered to the clinical staff to support and facilitate the implementation of the new TDM. The extent of the utilization of knowledge and implementation of the TDM varied across the three tertiary care mental health facilities. Focus groups (N=49) with clinical staff were conducted to explore various factors related to the training process as well as the challenges and benefits of implementing the TDM. Data were analyzed using Leininger's Phases of Ethnonursing Qualitative Data Analysis [Leininger, M. (2002). The theory of culture care and the ethnonursing research method. In M. Leininger & M. McFarland (eds.), Transcultural nursing: Concepts, theories, research, and practice (3rd ed.). New York: McGraw-Hill]. Findings revealed that clinical staff experienced challenges in roles and responsibilities, relationships with others, values and beliefs of clients, staff and community, resources, and the processes of care. From the findings, strategies were identified to support the integration of knowledge about the TDM into practice and to increase clinical staff's skills in implementing research interventions. These included support from others, as well as support of practice, policy, and education. PMID- 17397693 TI - Using data to increase the national presence of psychiatric mental health nursing. PMID- 17397694 TI - Accurate differential diagnosis: more than an algorithm. PMID- 17397695 TI - The test of our progress: affordable housing for seriously mentally ill clients. PMID- 17397696 TI - Story inquiry: a method for research. PMID- 17397697 TI - Helping veterans return: community, family, and job. PMID- 17397698 TI - A phase III randomized, double-blind, placebo-controlled study evaluating dextromethorphan plus slow-release morphine for chronic cancer pain relief in terminally ill patients. AB - This multicenter trial examined the efficacy and safety of dextromethorphan (DM) as an enhancer of analgesia and modulator of opioid tolerance in cancer patients with pain. Eligible patients were randomized to slow-release morphine plus DM or slow-release morphine plus placebo. The initial DM dose was 60 mg four times daily for seven days, with an increase to 120 mg four times daily, if tolerated, for another seven days. During the study, patients recorded medications and scores for pain, nausea, drowsiness, and insomnia. Sixty-five patients were randomized. Although average pain scores (12.6 vs. 15.8), number of breakthrough doses (9 vs. 11.3), and change in total morphine consumption (550.9 mg vs. 597.1mg) were less in the DM group than placebo group, the differences were not statistically significant (P=0.31-0.33). Side-effect scores were not statistically significantly different. Dizziness was greater in the DM (58%) than placebo (36%) group. This study showed a statistically nonsignificant enhancement of analgesia or modulation of opioid tolerance in cancer patients with pain when DM was added to morphine. Participants receiving the DM also had more toxicity, particularly dizziness. This toxicity and the limited evidence of effect do not support the use of DM to enhance opioid analgesia or to modulate opioid tolerance in cancer patients. PMID- 17397699 TI - Healing connections: on moving from suffering to a sense of well-being. AB - Life-threatening illness is an assault on the whole person--physical, psychological, social, and spiritual. It frequently presents caregiver and sufferer with a paradox--suffering does not correlate with physical well-being alone. Drawing on a purposive sample of 21 participants, a phenomenological study was carried out to explore the relevance of the existential and spiritual domains to suffering, healing, and quality of life (QOL). The phenomenological method was used to achieve an in-depth description of both existential suffering, and conversely, the experience of integrity and wholeness, in persons with life threatening illness; identify "inner life" and existential contributors to suffering and subjective well-being in advanced illness; and develop a narrative account of these QOL extremes. The importance of meaning-based adaptation to advanced illness was supported, as were Frankl's sources of meaning and Yalom's sources of existential anguish. Divergent themes characteristic of the two QOL extremes were identified. Four types of "healing connections" involving a sense of bonding to Self, others, the phenomenal world, and ultimate meaning, respectively, were identified. They situated the participant in a context that was greater and more enduring than the self, thus leading to enhanced meaning and QOL. The assumptions underlying the construct "health-related QOL" are questioned. PMID- 17397700 TI - Women experience higher levels of fatigue than men at the end of life: a longitudinal home palliative care study. AB - Few studies have evaluated sex differences in the prevalence, severity, and correlates of fatigue at the end of life. The Brief Fatigue Inventory, McGill Quality of Life (MQOL) Questionnaire, and Karnofsky Performance Scale were administered at two-week intervals to 102 patients in a home palliative program. Outcomes in the sample and a regional palliative database (n=3,096) were analyzed. Cancer was the diagnosis in 96% of patients enrolled. Prevalence (P=0.0091) and severity of fatigue (P<0.001) were higher in women at entry and in a repeated measures analysis over time (severity, P=0.0048). Performance status did not explain this difference. MQOL scores were inversely correlated to fatigue (Spearman coefficient=-0.48, P<0.0001), but did not differ by sex. There was no difference in fatigue interference with MQOL in women and men. Although depression was higher in women (P=0.042) and related to fatigue at entry, it did not explain the sex difference in fatigue scores. Of the sociodemographic variables examined, neither education nor living situation contributed to the fatigue difference. This study shows a sex effect in the fatigue experienced by patients with advanced illnesses, which is not explained by baseline differences in performance, depression, MQOL, education, or living situation. That fatigue interference with MQOL is the same for men and women suggests that higher fatigue scores in women reflect not only a difference in the dimension of fatigue severity, but are also relevant in relation to impact on QOL. Assessment of fatigue should include the dimension of QOL important for both women and men. PMID- 17397701 TI - Values of sleep/wake, activity/rest, circadian rhythms, and fatigue prior to adjuvant breast cancer chemotherapy. AB - Fatigue is the most prevalent and distressing symptom experienced by patients receiving adjuvant chemotherapy for early stage breast cancer. Higher fatigue levels have been related to sleep maintenance problems and low daytime activity in patients who have received chemotherapy, but knowledge describing these relationships prior to chemotherapy is sparse. The Piper Integrated Fatigue Model guided this study, which describes sleep/wake, activity/rest, circadian rhythms, and fatigue and how they interrelate in women with Stage I, II, or IIIA breast cancer during the 48 hours prior to the first adjuvant chemotherapy treatment. The present report describes these variables in 130 females, mean age=51.4 years; the majority were married and employed. Subjective sleep was measured by the Pittsburgh Sleep Quality Index and fatigue was measured by the Piper Fatigue Scale. Wrist actigraphy was used to objectively measure sleep/wake, activity/rest, and circadian rhythms. Mean Pittsburgh Sleep Quality Index score was 6.73+/-3.4, indicating poor sleep. Objective sleep/wake results were within normal limits established for healthy individuals, except for the number and length of night awakenings. Objective activity/rest results were within normal limits except for low mean daytime activity. Circadian rhythm mesor was 132.3 (24.6) and amplitude was 97.2 (22.8). Mean Piper Fatigue Scale score was 2.56+/ 2, with 72% reporting mild fatigue. There were significant relationships between subjective and objective sleep, but no consistent patterns. Higher total and subscale fatigue scores were correlated with most components of poorer subjective sleep quality (r=0.25-0.42, P< or =0.005). PMID- 17397702 TI - Transcutaneous electrical nerve stimulation vs. transcutaneous spinal electroanalgesia for chronic pain associated with breast cancer treatments. AB - Chronic pain associated with breast cancer treatment is becoming increasingly recognized. Patients with this condition can experience significant physical and psychological morbidity and may benefit from nonpharmacological interventions as part of a multidisciplinary team approach. We compared the effectiveness of transcutaneous electrical nerve stimulation (TENS), transcutaneous spinal electroanalgesia (TSE), and a placebo (sham TSE) in a randomized controlled trial. The study sample comprised 41 women with chronic pain following breast cancer treatment, and outcome measures included pain report, pain relief, pain interference, anxiety and depression, arm mobility, and analgesic consumption. There was little evidence to suggest that TENS or TSE were more effective than placebo. All three interventions had beneficial effects on both pain report and quality of life, a finding that may be due to either psychophysical improvements resulting from the personal interaction involved in the treatment or a placebo response. Although electrical stimulation appears to be well tolerated in this population, further research is needed to establish its effectiveness for chronic cancer treatment-related pain. PMID- 17397703 TI - Caregiver thoughts and feelings in response to different perspective-taking prompts. AB - Both conceptual and empirical evidence in the caregiving literature suggest that "how" informal caregivers are prompted to think about patient experiences can impact their ability to achieve perceptual agreement with patients on symptom events. Researchers have begun to test the effects of different clinical questions with caregivers or a proxy on their perceptual agreement with patient self-reports. However, there are gaps in understanding caregivers' underlying thoughts and feelings when they are prompted to take different vantage points on the patient's symptom experiences. To assess these thoughts and feelings, content analysis was performed on verbal responses to an open-ended interview schedule, in which caregivers were questioned about their thoughts and feelings in one of three randomly assigned instructional conditions-neutral, imagine-patient, and imagine-self perspective-taking. Responses were classified into one of five cue categories: patient-oriented, caregiver-oriented, generalized other, feeling distanced, and other. Caregivers tended to respond to patient symptom experiences in ways that suggest it is innate for caregivers in intimate relationships to make an effort to imagine the patient's viewpoint on symptom events. When caregivers were prompted to imagine the patient's perspective, "labeling" processes were also triggered that denote a categorization process in which caregivers interpreted the meaning of simple patient information arising in the care situation. A large portion of caregivers in the neutral and imagine-patient conditions reported feeling distanced from patients in light of perceived communication difficulties. PMID- 17397704 TI - Benefits of training family caregivers on experiences of closure during end-of life care. AB - Caregiving at Life's End (CGLE) is a program for family caregivers caring for someone during the last years of life that focuses on the emotional, spiritual, and practical aspects of life and relationship completion and closure. This study evaluated the effectiveness of CGLE in improving three major outcomes: comfort with caregiving, closure, and caregiver gain. Family caregivers (n=2,025) participated in programs facilitated by health and human service professionals (n=142) who completed a CGLE train-the-trainer workshop conducted by The Hospice Institute of the Florida Suncoast. The caregivers completed training rosters and pre- and/or post-surveys. Group differences are reported in baseline characteristics and change in three outcomes for caregivers who completed 1) both pre- and post-survey, 2) pre-survey only, and 3) post-survey only. For those who completed both surveys (n=926), paired t-tests and multiple linear regression tested the impact of program length on caregiver outcomes. Caregivers participated in, on average, four sessions and 7.7 hours of training. The majority of caregivers were Caucasian (88%), female (81%), and on average, 60 years old. Significant improvement was found in all three outcomes (P<0.001). The program length made a difference for improvement in comfort with caregiving and closure but not in caregiver gain. Caregivers who are caring for someone during the last years of life benefit from a program that focuses on the life-changing or transformative aspects of caregiving in the last years of life, as well as practical aspects of caregiving. The ability to support caregivers in this relatively low impact intervention can be used in hospice and nonhospice settings. PMID- 17397705 TI - Hospice utilization in Taiwan by cancer patients who died between 2000 and 2004. AB - To facilitate utilization of hospice services, Taiwan uses the National Health Insurance (NHI) as a major policy instrument. To evaluate the effect of this policy on hospice utilization by cancer patients during their final year of life, a retrospective cohort study was conducted by linking individual patient-level data from the National Register of Deaths Database and the NHI claims database to examine changes in the rates of hospice utilization, durations of patient survival (DOS) after enrollment, and the rates of late referrals to hospice care from 2000 to 2004. Among the 103,097 cancer patients who died between 2000 and 2004, the rate of hospice utilization during their final year of life grew substantially from 5.5% to 15.4%. However, Taiwanese cancer patients were enrolled in hospice care close to death (median DOS ranged from 14 to 47 days). Except for the small proportion of patients who received both inpatient hospice care and hospice home care, one-third to one-fourth of cancer decedents died within 7 days after being enrolled in hospice care. Although the rate of late referrals to hospice care did not vary much over time, the mean DOS for hospice care changed significantly. Many Taiwanese cancer patients who could potentially benefit from hospice care do not receive it in time. Further research is warranted to investigate factors influencing hospice use and the timing of hospice referrals to facilitate appropriate use of hospice care for cancer patients in Taiwan. PMID- 17397706 TI - Psychometric properties of the Impact of Event Scale in Greek cancer patients. AB - To develop the Greek version of the Impact of Event Scale-Revised (IES-R-Gr), assess its psychometric properties, and finally to examine the impact of cancer diagnosis to a palliative care patient sample, the IES-R was translated into Greek using the "forward-backward" procedure. It was administered twice, at one week intervals, to 82 eligible patients with advanced cancer. Together with the IES-R-Gr scale, the patients also completed the Hospital Anxiety and Depression (HAD) Scale. Reliability was assessed in terms of internal consistency (Cronbach's alpha coefficients) and test/retest (Spearman's r value and Kendall's tau-b). Construct validity was demonstrated through association with the HAD Scale, and convergence and discriminative validity and interscale correlations were also assessed. The Greek version of the IES-R had Cronbach's alphas for the intrusion, avoidance, and hyperarousal scales of 0.72, 0.77, and 0.85, respectively. Overall test-retest reliability was satisfactory at P<0.0005. Satisfactory construct validity was supported by the correlation analysis between the IES-R-Gr subscales and anxiety and depression. Factor analysis yielded three factors, explaining 57.26% of the variance. Interscale and interitem correlations were found satisfactory at P<0.0005. These results demonstrate that the IES-R-Gr is an instrument with satisfactory psychometric properties and is a valid research tool for the impact of cancer diagnosis in advanced cancer patients. PMID- 17397707 TI - Managing metastatic bone pain: the role of bisphosphonates. AB - Approximately two-thirds of patients with bone metastases have severe and debilitating pain. Despite a range of treatments, about 25% of patients with painful bone metastases suffer from uncontrolled pain. Bisphosphonates are the standard care for the reduction of skeletal events associated with bone metastases. We review the efficacy of currently available bisphosphonates in cancer-related bone pain. Oral clodronate, intravenous (i.v.) pamidronate, and i.v. zoledronic acid have shown an analgesic effect in some studies. Both i.v. and oral ibandronate reduced bone pain in breast cancer patients with bone metastases and maintained bone pain scores below baseline levels for up to two years in clinical trials. Pilot studies of intensive i.v. ibandronate dosing show rapid and effective relief from moderate-to-severe bone pain in patients with breast cancer and other tumors. Phase III trials are warranted to compare the efficacy of bisphosphonates in treating bone pain and to confirm the effects of intensive dosing regimens. PMID- 17397708 TI - Symptomatic therapy of dyspnea with strong opioids and its effect on ventilation in palliative care patients. AB - This study assessed the effect of opioid treatment on ventilation in dyspneic palliative care patients who received symptomatic treatment with strong opioids. The assessments measured changes in peripheral arterial oxygen saturation (SaO(2)), transcutaneous arterial pressure of carbon dioxide (tcPCO(2)), respiratory rate (f), and pulse rate (PF) during the titration phase with morphine or hydromorphone. The aims of the study were to verify the efficacy of opioids for the management of dyspnea, assess the effect on ventilation, and show whether nasal O(2) insufflation before opioid application leads to a decrease in the intensity of dyspnea. Eleven patients admitted to our palliative care unit were included in this prospective, nonrandomized trial. At admission, all patients suffered from dyspnea. tcPCO(2), SaO(2), and PF were measured transcutaneously by means of a SenTec Digital Monitor (SenTec AG, Switzerland). During O(2) insufflation, the intensity of dyspnea did not change. In contrast, the opioid produced a significant improvement in the intensity of dyspnea (P=0.003). Mean f decreased as early as 30 minutes after the first opioid administration, declining from 41.8+/-4.7 (35.0-50.0) to 35.5+/-4.2 (30.0-40.0), and after 90 minutes, to 25.7+/-4.5 (20.0-32.0) breaths/min. Other monitored respiratory parameters, however, showed no significant changes. There was no opioid-induced respiratory depression. PMID- 17397710 TI - Prophylaxis of postoperative endophthalmitis. PMID- 17397712 TI - Posterior capsule rupture after vitreoretinal surgery. PMID- 17397713 TI - Intraoperative floppy-iris syndrome with doxazosin. PMID- 17397714 TI - Association between clear corneal cataract incisions and postoperative endophthalmitis. PMID- 17397715 TI - Surgical management of acute angle-closure glaucoma after implantation of a toric ICL. PMID- 17397716 TI - Consultation section. Cataract surgical problem. PMID- 17397725 TI - Simple test to confirm cleavage with air between Descemet's membrane and stroma during big-bubble deep anterior lamellar keratoplasty. AB - We describe a simple test to confirm big-bubble formation in deep anterior lamellar keratoplasty by observing the position and movements of small air bubbles injected into the anterior chamber through a limbal paracentesis. The test also allows evaluation of the extension of Descemet's membrane cleavage from the posterior stroma relative to the margins of the corneal trephination. PMID- 17397726 TI - Scleral fixation in eyes with loss of capsule or zonule support. AB - We describe a new scleral fixation technique for posterior chamber intraocular lens (IOL) implantation in eyes with partial or total loss of the posterior capsule or zonule support. This technique uses a scleral incision that can be completed in less time than a conventional scleral flap and prevents unnecessary trauma to the eye. A double-thread, 10-0 polypropylene suture loop is introduced once through a scleral layer pathway inside the eye. The free ends of the polypropylene suture are buried between the 2 edges of the scleral incision. This technique was used in 15 eyes, with a follow-up of 6 to 30 months. Complications included iris capture, irregular pupil, hyphema, vitreous hemorrhage, choroidal hemorrhage, localized peripheral anterior synechias, and retinal detachment. The mean postoperative visual acuity was 20/40 at the last follow-up. This modified technique is an easy and effective way to achieve scleral fixation of the IOL. PMID- 17397727 TI - Microincision cataract surgery in a vitrectomized eye. AB - Phacoemulsification after vitrectomy is associated with several intraoperative problems. We describe the use of bimanual microincision cataract surgery using an 18-gauge needle as the irrigating chopper combined with silicone oil removal in a vitrectomized eye. This technique may offer a safe alternative to standard phacoemulsification in such cases. PMID- 17397728 TI - Combined preoperative topical atropine sulfate 1% and intracameral nonpreserved epinephrine hydrochloride 1:4000 [corrected] for management of intraoperative floppy-iris syndrome. AB - We describe a technique combining preoperative atropine sulfate 1% and intraoperative intracameral epinephrine in a 1:2500 dilution for the management of intraoperative floppy-iris syndrome (IFIS) induced by alpha(1A)-blocking agents such as tamsulosin. Patients on alpha(1A)-blocking agents used topical atropine sulfate 1% 3 times a day for 2 days before surgery. In addition to routine topical mydriatics before surgery, they received intracameral epinephrine diluted 1:2500 with BSS. In 19 of 20 eyes, there were no clinical manifestations of IFIS with this regimen. Stimulation of the iris dilator by a direct-acting sympathomimetic (epinephrine) combined with strong pupiloplegia (atropine) provides powerful synergism. Preoperative identification of patients on alpha(1) blockers is important as complications can be reduced significantly by using appropriate strategies to deal with this condition. It is also important that patients suffering from benign prostatic hyperplasia do not stop using an alpha(1A)-blocker, especially when preoperative atropine is used, as acute urinary retention may ensue. PMID- 17397729 TI - Long-term results of laser in situ keratomileusis for high myopia: risk for ectasia. AB - PURPOSE: To ascertain the long-term stability of laser in situ keratomileusis (LASIK) in highly myopic eyes. SETTING: Clinical practice office-based surgery. METHOD: Charts of eyes with high myopia who had LASIK surgery by the same surgeon between 1994 and 2000 were reviewed in 2003, and patients were given an appointment for follow-up examinations. In these highly myopic eyes, surgery was originally performed to create undercorrections with or without decreasing the ablation diameters to maximally conserve the residual stromal bed thickness. RESULTS: Of the 107 eyes with myopia between -10.00 diopters (D) and -35.00 D reviewed and operated on in a 3-year period between 1994 and 1998, 35 eyes of 31 patients had a single enhancement procedure. One case of ectasia as a result of excessive tissue removal occurred in a patient with a preoperative refraction of 28.00 D. Of the 107 eyes reviewed, 78 (73%) were examined after 5 years, 68 (63%) after 7 years, and 15 (14%) between 9 years and 11 years. CONCLUSIONS: Operating on eyes with highly myopic refractive errors and removing substantial tissue thickness did not produce ectasia in this series. Although high myopia has been considered a risk factor for post-LASIK ectasia, adherence to proper screening and intraoperative pachymetry appears to decrease the risk. PMID- 17397730 TI - Customized photoastigmatic refractive keratectomy using combined topographic and refractive data for myopia and astigmatism in eyes with forme fruste and mild keratoconus. AB - PURPOSE: To examine the outcomes of photoastigmatic refractive keratectomy using corneal and refractive parameters for myopia and astigmatism in eyes with forme fruste and mild keratoconus. SETTING: Private practice, Melbourne, Australia. METHODS: Photoastigmatic refractive keratectomy was performed with a Star 1 or Star 2 laser (Visx) in 45 eyes with forme fruste or mild keratoconus using the Alpins vector planning technique. Inclusion requirements were best corrected visual acuity (BCVA) 20/40 or better, no slitlamp signs of keratoconus, mean keratometry less than 50.00 diopters (D), and corneal and refractive stability for at least 2 years. RESULTS: Thirty-two eyes had follow-up of 5 years and 9 eyes, of 10 years. Preoperatively, the mean refractive astigmatism was -1.39 DC +/- 1.08 (SD) (range 0.45 to -5.04 DC) and the mean corneal astigmatism was 1.52 +/- 1.18 D (range 0.35 to 4.75 D) by manual keratometry and 1.70 +/- 1.42 D (range 0.32 to 5.32 D) by topography. Twelve months postoperatively, the mean refractive astigmatism was -0.43 +/- 0.40 D and the mean corneal astigmatism was 1.05 +/- 0.85 D by keratometry and 1.02 +/- 0.83 D by topography. At 12 months, the uncorrected visual acuity was 20/20 or better in 56% of eyes and 20/40 or better in all eyes. The BCVA was 20/20 or better in 89% of eyes and 20/30 or better in all eyes. Seven eyes had a loss of BCVA, and 16 eyes had a gain. There were no cases of keratoconus progression. CONCLUSIONS: Photoastigmatic refractive keratectomy in eyes with forme fruste and mild keratoconus was safe and effective for myopia and astigmatism in carefully selected patients with refractive and corneal stability. The incorporation of the corneal astigmatism data into the applied treatment parameters may improve visual and total astigmatism results. PMID- 17397731 TI - Contrast sensitivity and glare disability after implantation of AcrySof IQ Natural aspherical intraocular lens: prospective randomized masked clinical trial. AB - PURPOSE: To evaluate contrast sensitivity and glare disability after implantation of an AcrySof IQ Natural SN60WF aspherical intraocular lens (IOL) (Alcon Laboratories). SETTING: Iladevi Cataract & IOL Research Centre, Ahmedabad, India. METHODS: One hundred twenty consecutive patients who had phacoemulsification in a prospective triple-masked trial were randomized to receive an AcrySof SA60AT IOL (40 eyes), AcrySof Natural SN60AT IOL (40 eyes), or AcrySof IQ SN60WF IOL (40 eyes). At 3 months, contrast sensitivity was measured using the CSV-1000E contrast sensitivity chart test face (Vector Vision) at 3, 6, 12, and 18 cycles per degrees (cpd) under photopic conditions (85 cd/m(2)) and mesopic conditions (2.7 cd/m(2)) with 4.0 mm and 6.0 mm fixed central apertures, with and without glare. The Kruskal-Wallis test was used and a pair-wise comparison performed. The main outcome measure was the difference in contrast sensitivity between IOLs at each spatial frequency. RESULTS: The best corrected visual acuity was similar between the 3 IOL groups (P = .6). The AcrySof IQ group had significantly higher contrast sensitivity at 18 cpd under photopic conditions (P = .008); at 3, 6, 12, and 18 cpd during mesopic testing with a 4.0 mm aperture without glare (P = .018, P = .011, P = .007, and P = .0001, respectively) and with glare (P = .003, P = .006, P = .005, and P = .004, respectively); and at all spatial frequencies during mesopic testing with a 6.0 mm aperture without glare (P = .018, P = .006, P = .009, and P = .0001, respectively) and with glare (P = .019, P = .002, P = .01, and P = .017, respectively). CONCLUSION: Eyes with the AcrySof IQ SN60WF IOL had significantly higher contrast sensitivity than eyes with an AcrySof SA60AT or AcrySof Natural SN60AT IOL at all spatial frequencies during mesopic testing (with and without glare) with 4.0 and 6.0 mm artificial pupil. PMID- 17397732 TI - Bag-in-the-lens intraocular lens implantation in the pediatric eye. AB - PURPOSE: To study the efficacy, safety, and feasibility of implantation of a bag in-the-lens intraocular lens (IOL) in children and babies. SETTING: Departments of Ophthalmology, University Hospital, Antwerp, Belgium, and the University Hospital, Ljubljana, Slovenia, and a private ophthalmology practice, Oudenaarde, Belgium. METHODS: Thirty-four eyes of 22 children had implantation of a bag-in the-lens IOL. The ages ranged from 2 months to 14 years. Congenital cataract was present in 26 eyes, and persistent fetal vasculature (PFV) was concomitantly present in 4 eyes. Fifteen patients had bilateral cataract, and 6 had unilateral cataract. RESULTS: In 3 eyes, the IOL could not be properly implanted. In these cases, secondary intervention was necessary because of early posterior capsule opacification. The mean postoperative follow-up was 17.45 months +/- 17.12 (SD) (range 4 to 68 months). None of the children except those presenting with PFV had anterior vitrectomy during surgery. The optical axis remained clear during the follow-up in all patients who had successful IOL implantation. CONCLUSIONS: The bag-in-the-lens implantation technique in children and babies was safe and kept the visual axis clear after cataract surgery. In the near future, 4.0 or 4.5 mm IOLs will be available that may improve the success rate of IOL implantation in the small eyes of babies. PMID- 17397733 TI - Intraindividual comparison of surgical trauma after bimanual microincision and conventional small-incision coaxial phacoemulsification. AB - PURPOSE: To compare the surgical trauma after microincision phacoemulsification and small-incision coaxial phacoemulsification after implantation of conventional, foldable, hydrophobic acrylic intraocular lenses (IOLs). SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: A prospective investigator-masked case series comprised patients with bilateral cataract who had cataract surgery on the same day. Thirty-three patients (66 eyes) were randomized. Microincision cataract surgery (MICS) was performed through 2, 1.4 mm clear corneal incisions (CCIs) using bimanual sleeveless phacoemulsification (cool phaco) in 1 eye. Small-incision cataract surgery (SICS) was performed on the other eye through a 3.2 mm CCI. In all cases, an AcrySof SA60AT IOL was inserted, in the MICS group after the CCI was enlarged. Laser flare photometry, specular microscopy, corneal endothelial cell density, and pachymetry were evaluated preoperatively and postoperatively. Intraindividual comparison and statistical analyses were performed. RESULTS: There were no relevant clinical differences or perioperative complications in either group. There were no statistically significant differences between preoperative and postoperative anterior chamber flare or endothelial cell loss. On the first postoperative day, the MICS group had statistically significantly increased corneal swelling (P = .008). Postoperatively, the mean endothelial cell density loss was higher in the MICS group (6.2%) than in the SICS group (3.10%); however, the difference between groups was not significant (P = .08) CONCLUSIONS: Microincision cataract surgery was a safe and reproducible technique. The postoperative results in the MICS group were comparable to those in the SICS group. PMID- 17397734 TI - Effect of stromal hydration of clear corneal incisions: quantifying ingress of trypan blue into the anterior chamber after phacoemulsification. AB - PURPOSE: To use trypan blue as a quantifiable ingress tracer to determine whether stromal hydration reduces ocular surface fluid ingress at the end of phacoemulsification. SETTING: Iladevi Cataract and IOL Research Centre, Memnagar, Ahmedabad, India. METHODS: A prospective randomized study included 80 eyes having phacoemulsification through 2.2 mm incisions. These eyes were divided into 2 equal groups: 1 had stromal hydration (surgery completed by injecting fortified balanced salt solution [BSS Plus] to hydrate the lateral walls and internal entry of incision) and the other had no stromal hydration. One half milliliter of 0.0125% sterile trypan blue was instilled on the ocular surface and allowed to remain for 2 minutes. One-tenth milliliter of aqueous fluid was aspirated from the anterior chamber, and its optical density was measured using ultraviolet spectrophotometry. Logs of dilution of trypan blue were used for statistical analysis using the nonparametric Mann-Whitney U test. RESULTS: There was a statistically significant decrease and difference between groups in mean dilution of trypan blue in the aqueous aspirate (P<.001). The mean was 1:11,337 in the stromal hydration group and 1:220 in the no stromal hydration group. Logs of mean dilution of trypan blue had statistically significant lower values in the stromal hydration group than in the no stromal hydration group (3.21 and 2.14, respectively) (P<.001). CONCLUSIONS: Stromal hydration of clear corneal incisions reduced ingress into the anterior chamber of the trypan blue instilled on the ocular surface. Clinically, these findings may have a beneficial effect in reducing the risk for postoperative endophthalmitis. PMID- 17397735 TI - Effect of primary posterior continuous curvilinear capsulorhexis on clinical performance of ACR6D SE single-piece hydrophilic acrylic intraocular lenses. AB - PURPOSE: To evaluate the effects of a primary posterior continuous curvilinear capsulorhexis (PCCC) on the clinical performance of a single-piece hydrophilic acrylic intraocular lens (IOL) with haptic angulation. SETTING: Department of Ophthalmology, Medical University of Vienna, Vienna, Austria. METHODS: A prospective study of 52 patients with bilateral age-related cataract was conducted. Patients had standard cataract surgery with implantation of the same IOL in both eyes. Randomly, a PCCC was created in 1 eye and the posterior capsule was left intact in the fellow eye. Assessed parameters were visual axis opacification (VAO) in eyes with a PCCC or regeneratory posterior capsule opacification (PCO) in eyes without PCCC (scale 0 to 10) in the central (3.0 mm eccentricity), intermediate (3.0 to 4.5 mm eccentricity), and peripheral (>4.5 mm eccentricity [capsulorhexis edge]) areas; neodymium:YAG (Nd:YAG) laser capsulotomy or laser polishing of ongrowing lens epithelium; anterior capsule opacification (ACO); best corrected visual acuity (BCVA); and contrast sensitivity. RESULTS: Thirty patients completed the 2(1/2)-year follow-up. Visual axis opacification was significantly lower in the central region in the PCCC group (mean 0.5 +/- 0.7 [SD]) than PCO in the central region of the non-PCCC group (mean 1.1 +/- 1.1) (P = .02). Forty percent in the non-PCCC group had an Nd:YAG laser capsulotomy during the follow-up period; none in the PCCC group had laser polishing. There were no significant differences in ACO, SE, BCVA, or contrast sensitivity. There was no additional gain in BCVA or contrast sensitivity in eyes with a PCCC compared with eyes without a PCCC when VAO and PCO were low. CONCLUSION: A PCCC significantly reduced PCO formation within the central 3.0 mm eccentricity as well as the need for Nd:YAG laser capsulotomy in eyes with a single-piece hydrophilic acrylic IOL with angulated haptics. PMID- 17397736 TI - Polymerase chain reaction identification in aqueous humor of patients with postoperative endophthalmitis. AB - PURPOSE: To identify bacterial agents in the aqueous humor of patients with postoperative endophthalmitis using eubacterial polymerase chain reaction (PCR) and conventional culture. SETTING: University Hospital of Lyon E. Herriot, Lyon, France. METHODS: Broad-range eubacterial PCR amplification followed by direct sequencing was used to identify microbial pathogens in ocular samples from 30 patients with acute or delayed-onset endophthalmitis, mainly after cataract surgery. Ocular samples included aqueous humor collected before the first intravitreal injection of antibiotics and vitreous samples collected at the time of the therapeutic pars plana vitrectomy. RESULTS: Cultures were positive in 32% of cases and PCR in 61% of cases with aqueous humor samples. When associated, culture and PCR of aqueous humor samples allowed for a microbiological diagnosis in 71% of cases. Microorganisms cultured by conventional techniques matched those identified by PCR. When applied on vitreous pretreated with intravitreal antibiotics, PCR increased the identification rate from 18% to 62%. CONCLUSIONS: Polymerase chain reaction assay of initial aqueous humor samples contributed to the diagnosis of endophthalmitis in 30% of cases. Previous use of intravitreal antibiotics did not seem to affect the ability to PCR-amplify DNA in the short term. Polymerase chain reaction-based technology was a useful adjunct to conventional culture because when used with aqueous humor samples only, the association of both techniques allowed for a microbiological diagnosis in 71% of cases of postoperative acute and delayed-onset endophthalmitis. PMID- 17397737 TI - Effectiveness of laser-assisted subepithelial keratectomy to treat residual refractive errors after laser in situ keratomileusis. AB - PURPOSE: To evaluate the effectiveness of laser-assisted subepithelial keratectomy (LASEK) to treat residual refractive errors after laser in situ keratomileusis (LASIK). SETTINGS: Isik Eye Clinic, Ankara, Turkey. METHODS: This retrospective study included 24 eyes of 15 patients who had retreatment by LASEK for residual refractive errors after myopic LASIK. All patients had examinations that included slitlamp biomicroscopy, subjective and cycloplegic refractions, uncorrected visual acuity (UCVA), best corrected visual acuity, corneal topography, and pachymetry preoperatively and postoperatively. Postoperative examinations were performed at 1 week and 1, 3, and 6 months. RESULTS: The patient cohort comprised 9 men and 6 women. The median spherical equivalent (SE) of attempted correction for retreatment with LASEK was -1.25 diopters (D). The median follow-up after LASEK was 11.5 months (range 6 to 16 months). At the end of the follow-up, the median SE of the refractive error was -0.38 D. The median UCVA increased from 20/45 before LASEK to 20/25 at the last follow-up visit, which was statistically significant (P<.001). After LASEK, significant postoperative haze developed in 5 eyes. In all 5 eyes, the estimated ablation depth was more than 40 mum and the SE of attempted correction was -2.00 D or greater. CONCLUSIONS: Laser-assisted subepithelial keratectomy retreatment in eyes with myopic regression after LASIK resulted in a significant improvement in UCVA that was comparable to the improvement after flap lifting. An SE of attempted correction greater than -2.00 D was associated with a significant rate of haze. PMID- 17397738 TI - Comparison of outcomes of 2 channel sizes for intrastromal ring segment implantation with a femtosecond laser in eyes with keratoconus. AB - PURPOSE: To compare the outcomes of 2 channel sizes for intrastromal ring segment (Intacs, Addition Technology, Inc.) implantation using a femtosecond laser in eyes with keratoconus. SETTING: Kudret Eye Hospital, Ankara, Turkey. METHODS: A retrospective outpatient study included 159 consecutive keratoconic eyes of 103 patients. Patients were classified into 2 groups according to channel dissection size: wide (6.7 mm x 8.2 mm) and narrow (6.6 mm x 7.6 mm). Dissections were created with an IntraLase femtosecond laser (IntraLase Corp.). The 2 groups were compared according to uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refractive error (manifest sphere, manifest cylinder, and manifest refractive spherical equivalent), mean K-value, and complications within 6 months postoperatively. RESULTS: The wide-channel group comprised 97 eyes of 65 patients (37 men, 28 women) with a mean age of 27.64 years +/- 8.61 (SD). The narrow channel group comprised 62 eyes of 38 patients (23 men, 15 women) with a mean age of 26.62 +/- 6.13 years. At 6 months, the UCVA and BCVA had significantly improved in 63.9% and 70.1% of eyes, respectively, in the wide-channel group and in 72.5% and 75.8% of eyes, respectively, in the narrow-channel group. The change in mean BCVA and change in mean UCVA in both groups were not statistically significant (P<.05, independent samples t test). There was no between-group difference in the improvement in manifest spherical refraction, cylindrical refraction, manifest refractive spherical equivalent, or mean K-readings (P<.05, independent samples t test). Epithelial plug, yellow-white deposits, tunnel haze around segments, and upward movement of the inferior segment without extrusion were observed more frequently in the narrow-channel group than in the wide channel group. CONCLUSIONS: The refractive outcomes of wide channels and narrow channels for Intacs treatment in eyes with keratoconus were similar 6 months postoperatively. The narrow-channel group had a higher rate of mild to moderate complications than the wide-channel group. PMID- 17397739 TI - Zernike versus Fourier treatment tables for myopic patients having CustomVue wavefront laser in situ keratomileusis with the S4 excimer laser. AB - PURPOSE: To compare the efficacy, predictability, and safety of Zernike-based versus Fourier-based treatment tables in myopic eyes 1 year after CustomVue wavefront laser in situ keratomileusis (LASIK) using the S4 excimer laser (Visx). SETTING: Department of Ophthalmology, Stanford University School of Medicine, Stanford, California, USA. METHODS: A prospective analysis of 32 eyes of 16 patients with myopia who had CustomVue LASIK with the S4 excimer laser was conducted using Zernike or Fourier treatment tables. Mean spherical equivalent (SE), uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), and higher wavefront analyses were determined. RESULTS: The mean postoperative SE was -0.27 diopter (D) +/- 0.41 (SD) in Zernike-treated eyes and 0.20 +/- 0.29 D in Fourier-treated eyes. Twenty-eight of 32 eyes were available for analysis. Ninety-two percent of eyes in the Zernike group and all eyes in the Fourier group had 20/20 or better acuity (P = .85). Seventy-nine percent and 92%, respectively, were within +/-0.50 D of emmetropia (P = .68). All eyes in the Zernike group and 93% of eyes in the Fourier group had no change or gained at least 1 Snellen line of BSCVA (P = .85). Mean higher-order aberration values (mum) in the Zernike group and Fourier group were as follows: total root mean square, 0.91 and 0.99, (P = 0.95); defocus, 0.71 and 0.74, (P = 0.98); astigmatism, 0.15 and 0.31, (P = 0.81); coma, 0.21 and 0.20, (P = 0.99); trefoil, 0.11 and 0.11, (P = 1); and spherical aberration, 0.17 and 0.30, (P = 0.85), respectively. CONCLUSIONS: Myopic eyes that had CustomVue LASIK had better visual outcomes in the Fourier group than in the Zernike group. Higher-order wavefront aberration results were similar between the 2 groups. PMID- 17397740 TI - Effect of a yellow intraocular lens on scotopic vision, glare disability, and blue color perception. AB - PURPOSE: To compare the photopic and scotopic contrast sensitivity with and without glare as well as blue color perception between eyes with an AcrySof SN60AT Natural intraocular lens (IOL) (Alcon Laboratories Inc.) and eyes with a conventional AcrySof SA60AT IOL. SETTING: Ankara University School of Medicine, Department of Ophthalmology, Ankara, Turkey. METHODS: Right eyes of 38 patients with an AcrySof Natural IOL and right eyes of 38 age-matched patients with a conventional AcrySof SA60AT IOL were included in a study. Contrast sensitivity was measured with the Functional Acuity Contrast Test under photopic conditions. Scotopic contrast sensitivity in the presence or absence of glare was measured using the Mesotest II (Oculus GmbH). Blue-green color vision was evaluated with the Moreland equation of the HMC Anomaloskop MR (Oculus GmbH). RESULTS: The mean age of patients was 66.6 years +/- 8.2 (SD) in the Natural IOL group and 66.4 +/- 8.0 years in the conventional IOL group. There was no statistically significant difference in photopic contrast sensitivity, scotopic contrast sensitivity with and without glare, or disability glare between the 2 groups (P>.05). Photopic and scotopic contrast sensitivity with and without glare significantly decreased with age in both groups (P<.01). There was no statistically significant difference in anomaloscope scores between the 2 groups (P>.05). The Moreland middle match point showed a significant shift toward blue with age in both groups (P<.01). CONCLUSIONS: The AcrySof SN60AT Natural IOL provided contrast sensitivity under photopic and scotopic conditions (with and without glare) and blue color perception comparable that obtained with the AcrySof SA60AT IOL. Scotopic vision and blue color discrimination decreased with age with both IOLs. PMID- 17397742 TI - Comparison of the corneal response to laser in situ keratomileusis with flap creation using the FS15 and FS30 femtosecond lasers: clinical and confocal microscopy findings. AB - PURPOSE: To compare the response of the cornea to laser in situ keratomileusis (LASIK) with flap creation using the IntraLase FS15 or FS30 femtosecond laser (IntraLase Corp.). SETTING: Department of Ophthalmology, University of Texas Southwestern Medical Center, Dallas, Texas, USA. METHODS: Twenty-three patients (31 eyes) who had LASIK with flap creation using the FS15 or FS30 laser were assessed by clinical examination and confocal microscopy in a nonrandomized parallel treatment group comparative trial. Eight FS15 patients (15 eyes) were examined preoperatively and 3 months postoperatively, and 14 FS30 patients (15 eyes) were examined 3 months postoperatively. RESULTS: No patient in either group had clinically significant flap interface haze. One FS15 eye and 1 FS30 eye had significant keratocyte activation at the flap interface. The mean difference between the actual flap thickness and intended flap thickness was 16.8 microm +/- 11.1 (SD) and 13.9 +/- 7.1 microm in the FS15 group and FS30 group, respectively (P = .49). The mean measured interface reflectivity was 156.4 +/- 88.6 confocal backscatter units (CBU) and 104.8 +/- 91.2 CBU, respectively (P = .15). The mean density of interface particles was 21.4 +/- 14.8 particles/mm(2) in the FS15 group and 11.0 +/- 7.1 particles/mm(2) in the FS30 group (P<.05). CONCLUSIONS: Both the FS15 and FS30 lasers provided more reproducible flap thickness and fewer interface particles than previously observed using microkeratomes. The response of corneal keratocytes to intra-LASIK was reduced compared with previous results in which higher raster energies were used. Compared with the FS15, there was an apparent reduction in overall interface reflectivity and fewer interface particles with the FS30 laser. PMID- 17397741 TI - Scotopic sensitivity and color vision with a blue-light-absorbing intraocular lens. AB - PURPOSE: To investigate possible adverse effects of a yellow-tinted intraocular lens (IOL) on scotopic sensitivity and hue discrimination. SETTING: Departments of Ophthalmology, Columbia University and New York University School of Medicine, New York, New York, USA. METHODS: Nine patients with a yellow-tinted IOL in 1 eye and a colorless ultraviolet IOL in the fellow eye and 9 young phakic subjects with and without a yellow-tinted clip-on lens were tested. Hue discrimination was measured with the Farnsworth-Munsell (FM) 100-hue test. Dark-adapted thresholds to 440 nm, 500 nm, and 650 nm lights were measured at 23 locations using a modified Humphrey perimeter, and dark-adapted thresholds to white light were measured at 15 degrees temporal retina. RESULTS: In the 9 patients, there were no significant differences in dark-adapted sensitivities to 440, 500, 650 nm, or white-light stimuli and no differences in FM 100-hue error scores between eyes with yellow-tinted IOLs and those with colorless IOLs. Similarly, in young phakic subjects, there were no significant differences in FM 100-hue error scores or dark-adapted sensitivity to the white light with and without the yellow-tinted clip-on lens. However, with the clip-on lens, mean sensitivities to the 440 nm, 500 nm, and 650 nm stimuli were significantly decreased by 2.7 to 2.8 dB, 0.7 to 1.0 dB, and 0 to 1.2 dB, respectively. CONCLUSION: Results suggest that implantation of a yellow-tinted IOL has non-significant effect on scotopic sensitivity and hue discrimination. PMID- 17397743 TI - Successful treatment of cystoid macular edema with valdecoxib. AB - PURPOSE: To evaluate the safety and efficacy of the COX-2 inhibitor valdecoxib in treating macular edema after cataract surgery. SETTING: University Eye Clinic, Freiburg, Germany and Reis Medical Institution, Liechtenstein. METHODS: The COX-2 inhibitor valdecoxib (Bextra) was administered systemically to patients with significant visual loss resulting from macular edema in a prospective clinical trial. RESULTS: Ten patients were enrolled. Valdecoxib was tolerated well and led to a significant visual improvement within 10 days of therapy in all patients. CONCLUSION: The fast and persistent control of macular edema with valdecoxib warrants further investigation. PMID- 17397744 TI - Validity of the keratometric index: large population-based study. AB - PURPOSE: To determine the accuracy of the keratometric index of 1.3315 based on the Gullstrand model eye in predicting the power of the posterior cornea, Gullstrand's model was compared to a calculated keratometric index derived from actual measurements of the cornea. SETTING: Eye Institute, Tan Tock Seng Hospital, Singapore. METHODS: One eye of 2429 subjects with a mean spherical equivalent of -5.32 diopters (D) +/- 2.88 (SD) was measured with the Orbscan II (Bausch & Lomb). The following variables were analyzed: anterior radius of curvature (r(anterior)), posterior radius of curvature (r(posterior)), radius of keratometry (r(simK)), and central pachymetry. RESULTS: The r(anterior), r(posterior), and r(simK) were normally distributed, with a mean of 7.87 +/- 0.25 mm (95% confidence interval [CI], 7.38-8.36), 6.46 +/- 0.26 mm (95% CI, 5.95 6.97), and 7.71 +/- 0.27 mm (95% CI, 7.18-8.24), respectively. The mean ratio between the anterior corneal curvature and posterior corneal curvature was 1.22 +/- 0.03 (95% CI, 1.16-1.28). Based on the measurements of each eye, the mean calculated keratometric index, N(calc), was 1.3273 +/- 0.0013 (95% CI, 1.3248 1.3298). Using N(calc), the posterior corneal power was predicted to within +/ 0.50 D of the actual posterior power in 98.3% of eyes. The mean absolute error between the actual and calculated posterior power was 0.157 +/- 0.123 D using N(calc) and 0.326 +/- 0.133 D using the Gullstrand model. CONCLUSION: Modifying the keratometric index increased the accuracy of predicting the posterior corneal power. PMID- 17397745 TI - Repositioning and scleral fixation of the subluxated lens capsule using an intraocular anchoring device in experimental models. AB - PURPOSE: To describe a new device for repositioning and scleral fixation of the capsular bag of subluxated lenses in experimental models. SETTING: Laboratory for Experimental Microsurgery, Meir Medical Center, Kfar-Saba, Israel. METHODS: The capsular anchoring device is a poly(methyl methacrylate) 1-plane implant consisting of 2 handles that grasp the edges of the capsulorhexis and a base for scleral fixation with a single 10-0 or 9-0 polypropylene suture. A temporary safety suture can be used to facilitate manipulations and prevent loss of the device through the zonular defect until it is secured to the scleral wall. The device was implanted in porcine eyes and living rabbit eyes. An animal model of lens subluxation was achieved by tearing about one third of the zonules. Capsule centration, implant stability, and inflammatory reaction were evaluated 2 to 4 weeks after implantation in the living model. RESULTS: The device was implanted in 7 porcine eyes and 9 living rabbit eyes. A lens subluxation model was created in 4 porcine eyes and 2 rabbit eyes. The device effectively pulled the capsular bag to the center and remained stable up to 4 weeks thereafter. Very large zonular dialysis (>5 hours) was managed using 2 devices. Successful intraocular lens implantation was done repeatedly in the presence of the device. The implant was well tolerated in all rabbit eyes. Histopathological examination of the enucleated eyes revealed no inflammatory reaction or adhesions. The margin of the anterior capsulorhexis remained intact in all cases. CONCLUSIONS: Experimental studies of a capsular anchoring device for subluxated lenses confirmed the safety and efficacy of the new device. A capsular tension ring can also be inserted separately to further stabilize the capsular bag. PMID- 17397746 TI - In situ ablation of lens epithelial cells in porcine eyes with the laser photolysis system. AB - PURPOSE: To evaluate the efficacy of the laser photolysis system (LPS) (A.R.C. Laser GmbH) in removing lens epithelial cells (LECs) to prevent posterior capsule opacification (PCO) in an in situ model. SETTING: Department of Ophthalmology, Friedrich-Alexander-University Erlangen-Nurnberg, Erlangen, Germany. METHODS: Twelve enucleated porcine eyes fixed in a specially developed eye holder were randomly assigned to the control or treatment group. The cornea and iris were removed from all eyes, and a small paracentral capsulorhexis was performed. The lens nucleus and cortex were extracted by hydroexpression. The tip of the LPS was inserted into the capsular bag of eyes in the treatment group, and 50 pulses (10 mJ) were applied to the anterior capsule. All capsules were evaluated for remaining LECs by confocal laser scanning microscopy (HRT II with the Rostock Cornea Module, Heidelberg Engineering) and standard histology (hematoxylin-eosin and periodic acid-Schiff stains). RESULTS: In the control group, a homogenous layer of LECs attached to the anterior capsule was seen with both evaluation methods. In the treatment group, no LECs adherent to the anterior capsule were detected, suggesting complete ablation of LECs from the capsule. Small islands of equatorial LECs were found in places in which the remaining cortical fibers protected cells from the laser shockwave. The results of the confocal laser scanning microscopy were confirmed by standard histology. CONCLUSIONS: The LPS completely ablated LECs in an in situ model of cataract extraction. This system might prevent formation of PCO in vivo. PMID- 17397747 TI - In vitro efficacy of newly designed vancomycin-based microparticles. AB - PURPOSE: To compare the in vitro bactericidal and anti-adhesion properties of vancomycin-based microparticles and lyophilized vancomycin and estimate their relevance to perioperative antibiotic prophylaxis and endophthalmitis prevention. SETTING: University research laboratory, Lyon, France. METHODS: The bactericidal and anti-adhesion properties of a newly designed drug-delivery system were assessed on Staphylococcus epidermidis clinical strain N890074 containing the intercellular adhesion locus ica. Lyophilized vancomycin at 20 mug/mL was used as a standard. The new drug-delivery system, designed for the study, consisted of sterile, biocompatible, and biodegradable microparticles with continuous release of vancomycin. To obtain bacterial killing and anti-adhesion curves, experiments were first performed in a bacterial suspension containing 1000 colony-forming units per milliliter. Experiments were then performed with intraocular lenses incubated in the suspension. Efficacy was investigated by bacterial counts and scanning electron microscopy observations. RESULTS: The bactericidal and anti adhesion effects of vancomycin-based microparticles started after 3 hours (P<.002) and 1 hour (P<.001), respectively, and of lyophilized vancomycin, after 1 hour (P = .004) and 1 hour (P<.001), respectively. There was no difference between the 2 forms of vancomycin in the bactericidal effect starting at 21 hours and the anti-adhesion effect starting at 6 hours (P>.05). CONCLUSIONS: The newly designed vancomycin-based microparticles showed relevant antibacterial and anti adhesion properties after releasing a sufficient antibacterial quantity, proving that vancomycin remains efficient after undergoing the encapsulation process. PMID- 17397748 TI - Noninvasive identification of different intraocular lenses using Raman spectroscopy in porcine eyes. AB - PURPOSE: To develop a noninvasive technique for identifying the specific type of intraocular lens (IOL) implanted in porcine eyes. SETTING: Department of Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, and Department of Chemistry Education, Seoul National University, Seoul, Korea. METHODS: A confocal Raman spectroscopy system was used to identify various acrylic (AcrySof SA60AT [Alcon], Sensar AR40e [Advanced Medical Optics], I-Flex [I-Medical Intraocular Lenses]) and silicone (ClariFlex, [Advanced Medical Optics]) IOLs implanted in enucleated pig eyes. Confocal Raman spectra of IOLs were obtained by photoexcitation using the 514.5 nm line from an argon-ion laser, with a power of 1 mW at the sample and an acquisition time of 10 to 100 seconds. RESULTS: All IOLs, including those composed of acrylic optic material from different manufacturers, exhibited distinct spectral peaks before and after implantation in pig eyes. CONCLUSION: Confocal Raman spectroscopy may represent a novel method for obtaining specific information about implanted IOLs through direct and noninvasive confocal observation of spectral characteristics. PMID- 17397749 TI - Causes of intraocular lens opacification or discoloration. AB - Various pathologic processes may lead to clinically significant opacification or discoloration of the optic component of intraocular lenses (IOLs) manufactured from different biomaterials and in different designs. Factors such as the patient's associated conditions, the manufacturing process, the method of IOL storage, the surgical technique and adjuvants, or a combination of these may be involved. The complication may be observed intraoperatively or postoperatively from a few hours after implantation to many years after surgery, depending on the processes involved. Based on a review of the literature as well as our own laboratory analyses, the following types of processes were identified: formation of deposits/precipitates on the IOL surface or within the IOL substance; opacification by excess influx of water in hydrophobic materials; direct discoloration by capsular dyes or medications; coating by substances such as ophthalmic ointment and silicone oil; and a slow, progressive degradation of the IOL biomaterial. PMID- 17397750 TI - Primary piggyback implantation of 3 intraocular lenses in nanophthalmos. AB - We present a patient with bilateral nanophthalmos who had uneventful cataract extraction in the right eye with primary implantation of 3 intraocular lenses (IOLs) of 2 different materials: a 30 diopter (D) acrylic IOL and a 9 D silicone IOL in the capsular bag and a 30 D silicone IOL in the ciliary sulcus. Subsequently, cataract extraction was done in the left eye with bag-sulcus implantation of two 30 D silicone IOLs. The use of 3 IOLs in 1 eye was necessary because the highest available power of acrylic and silicone IOLs at our institution was 30 D. The only short-term complications were temporary corneal edema and partial displacement of the sulcus IOL anterior to the iris in the right eye and bilateral posterior capsule opacification. The late complication of interlenticular opacification was not present 1 year after piggyback IOL implantation. PMID- 17397751 TI - Postoperative infectious endophthalmitis after triamcinolone-assisted anterior vitrectomy. AB - Extracapsular cataract extraction and intraocular lens (IOL) implantation was performed in the left eye of a 64-year-old woman. Preoperative assessment showed a hypermature cataract in the left eye with visual acuity of hand movement. During IOL implantation, a posterior capsule rupture occurred in the inferior area. Triamcinolone was injected into the anterior chamber to visualize the vitreous and helped in thorough removal of the prolapsed vitreous. However, when postoperative infectious endophthalmitis developed, the signs and symptoms were vague. Careful examination, specific patient instructions, and close follow-up are necessary after a triamcinolone-assisted anterior vitrectomy. PMID- 17397752 TI - Postoperative cilium entrapment by clear corneal incision. AB - A 75-year-old man had routine phacoemulsification cataract extraction with posterior chamber intraocular lens implantation in the left eye using a temporal corneal incision and inferior paracentesis. Examinations at 1 day and 1 week were unremarkable; however, at the 6-week assessment, a cilium was noted to have penetrated the external ostium of the paracentesis, with the proximal (follicle) end abutting the internal ostium of the wound. While the cilium was removed without incident, this chance finding may aid our understanding of how intraocular cilia are occasionally discovered following routine small-incision sutureless cataract surgery. PMID- 17397753 TI - Surgical approach, histopathology, and pathogenesis in cataract associated with true lens exfoliation. AB - We report a case of true exfoliation of the crystalline lens in a 53-year-old baker who presented with reduced vision due to cataract. Phacoemulsification cataract surgery with intraocular lens implantation was performed, and the anterior capsule was analyzed histologically. Preoperative clinical and anterior segment optical coherence tomography observations are presented, with a description of the modified phacoemulsification technique and histopathology. A possible mechanism of pathogenesis is proposed in the context of the histopathology. PMID- 17397754 TI - Posttraumatic Aspergillus terreus endophthalmitis masquerading as dispersed lens fragments. AB - A 20-year-old man developed increasing inflammation with a hypopyon 3 weeks after primary repair of a corneal laceration. An occult anterior capsule puncture was suspected as the stimulus for the inflammation. When aspiration of the suspected hydrated lens material was not curative, a vitrectomy with injection of intravitreal antibiotic agents including amphotericin (0.0125 mg) was done the following day; the culture failed to grow fungal organisms. A repeat vitrectomy was performed 1 week later, and the culture grew Aspergillus terreus. This was determined to be resistant to amphotericin so voriconazole was injected intravitreally. The inflammation recurred, and the eye required enucleation because of blindness and intractable pain. Fungal endophthalmitis should be considered in cases of delayed-onset inflammation after trauma and may be due to organisms resistant to amphotericin. PMID- 17397755 TI - Pigmented free-floating retrolental space cyst. AB - We report an unusual case of a free-floating pigmented retrolental cyst, which was diagnosed after examination by slitlamp, B-scan ultrasonography, and ultrasound biomicroscopy. Pigmented cysts, which may arise from the ciliary body epithelium, are embryologically and morphologically different from clear cysts. PMID- 17397756 TI - Early opacification and subsequent clearing of a MemoryLens intraocular lens. AB - A 74-year-old woman had cataract extraction with implantation of a MemoryLens posterior chamber intraocular lens (IOL) (model CV232, Ioltech). On the first postoperative day, she complained of hazy vision and an opaque area was found in the IOL. Explantation was scheduled but postponed when the patient's vision improved and the IOL cleared by the second postoperative day. To date, the IOL has remained clear. To our knowledge, this is the first report of resolving opacification in a MemoryLens. Two cases of opacification in the same IOL model have been reported; the opacification persisted, and the IOLs were explanted. In this case, early opacification and resolution suggest an acute process. With early IOL opacification, it may be beneficial to delay explantation for a few days. PMID- 17397757 TI - Anorexiant-induced transient myopia after myopic laser in situ keratomileusis. AB - We report a case of acute transient myopia associated with ciliochoroidal effusion induced by anorexiants. The patient had had myopic laser in situ keratomileusis 7 years earlier. Acute bilateral myopia associated with anterior chamber shallowing, intraocular pressure elevation, diffuse ciliochoroidal effusion, and perimacular retinal folds was relieved 14 days after discontinuation of anorexiant medications. Tropicamide and atropine were used to deepen the anterior chamber. Sympathomimetic drugs such as phendimetrazine and ephedrine are used as anorexiants and may induce transient myopia associated with ciliochoroidal effusion, shallow anterior chamber, and acute angle-closure glaucoma. PMID- 17397758 TI - The Sandwell technique of Sub-Tenon's infiltration anesthesia for cataract surgery and other ophthalmic procedures. PMID- 17397759 TI - Addendum to ultrasound axial length measurement in biphakic eyes: factors for Alcon L12500-L14000 anterior chamber phakic IOLs. PMID- 17397760 TI - Ex vivo production of human red blood cells from hematopoietic stem cells: what is the future in transfusion? AB - There is difficulty in obtaining adequate supplies of blood components, as well as disappointing performance of stabilized or recombinant hemoglobins, limited indications of oxygen transporters (perfluorocarbons), and slow development of "universal" red blood cells (RBCs). There is, therefore, a need for complementary sources of RBCs for transfusion. Thus, an attempt to generate erythroid cells in vitro makes good sense. We describe in this article a methodology permitting the massive ex vivo production of mature human RBCs having all the characteristics of native adult RBCs from hematopoietic stem cells of diverse origins: blood, bone marrow, or cord blood. This protocol allows both the massive expansion of hematopoietic stem cells/progenitors and their complete differentiation to the stage of perfectly functional mature RBCs. The levels of amplification obtained (10(5) to 2 x 10(6)) are compatible with an eventual transfusion application. We discuss in this article the state of the art of this new concept and evoke possible obstacles that need to be overcome to pass from a laboratory model to clinical practice. We analyze its possible indications in the medium and long term, discuss the economic aspects, and raise the question: Can we afford the luxury of developing this approach, one that could represent a considerable advance in blood transfusion? PMID- 17397761 TI - Modern plasma fractionation. AB - Protein products fractionated from human plasma are an essential class of therapeutics used, often as the only available option, in the prevention, management, and treatment of life-threatening conditions resulting from trauma, congenital deficiencies, immunologic disorders, or infections. Modern plasma product production technology remains largely based on the ethanol fractionation process, but much has evolved in the last few years to improve product purity, to enhance the recovery of immunoglobulin G, and to isolate new plasma proteins, such as alpha1-protease inhibitor, von Willebrand factor, and protein C. Because of the human origin of the starting material and the pooling of 10,000 to 50,000 donations required for industrial processing, the major risk associated to plasma products is the transmission of blood-borne infectious agents. A complete set of measures--and, most particularly, the use of dedicated viral inactivation and removal treatments--has been implemented throughout the production chain of fractionated plasma products over the last 20 years to ensure optimal safety, in particular, and not exclusively, against HIV, hepatitis B virus, and hepatitis C virus. In this review, we summarize the practices of the modern plasma fractionation industry from the collection of the raw plasma material to the industrial manufacture of fractionated products. We describe the quality requirements of plasma for fractionation and the various treatments applied for the inactivation and removal of blood-borne infectious agents and provide examples of methods used for the purification of the various classes of plasma protein therapies. We also highlight aspects of the good manufacturing practices and the regulatory environment that govern the whole chain of production. In a regulated and professional environment, fractionated plasma products manufactured by modern processes are certainly among the lowest-risk therapeutic biological products in use today. PMID- 17397762 TI - Transfusion in the patient with sickle cell disease: a critical review of the literature and transfusion guidelines. AB - The clinical outcomes of sickle cell disease (SCD) have vastly improved over the years in great part as a result of advanced medical technologies, improved patient education, and multidisciplinary care. A key component in the successful management of patients with SCD is red blood cell transfusion therapy used in the treatment and prevention of sickle cell complications. However, although the successful application of transfusion therapy has significantly improved the morbidity and mortality of patients with SCD, the literature that addresses the appropriate selection and use of blood products continues to evolve with no clear universal standard of care. Our objectives were to provide an in-depth review of the current literature on transfusion therapy in SCD and to provide a set of guidelines for the transfusion management of patients with SCD. PMID- 17397763 TI - Chimerism in the immunohematology laboratory in the molecular biology era. AB - Dual or multiple cell populations, induced by chimeras, have been the subject of many studies. This long-standing fascination with chimeras has revealed a good deal of knowledge about human inheritance. Although historically most chimeras were caused by natural events, certain current medical intervention therapies are increasing the number of situations that can lead to a mixed cell population, that is, the chimeric condition, in humans. Medical therapies such as transfusion, stem cell transplantation, kidney transplantation, and artificial insemination induce temporary and sometimes permanent chimeras. Such natural or therapeutically induced presentations of chimerism can present challenging issues to the clinical immunohematology laboratory with regard to interpretation of results and subsequent patient management. The purpose of this review was to highlight some of these chimeric states and hypothesize how testing DNA from various tissues can cause apparent discrepancies between phenotype and genotype results. PMID- 17397764 TI - The National Blood Service (England) approach to evaluation of kits for detecting infectious agents. AB - Blood services test their donations for a range of infectious agents before release for transfusion. To ensure that the assays used have appropriate sensitivity, subtype detection range, and specificity, and meet operational requirements (timeliness, automation, and process control), some form of selection is needed. The approach of the English National Blood Service (NBS) to the evaluation of commercial kits to assess their suitability is presented. As a centrally coordinated national service the NBS has the "critical mass" and can generate the economies of scale, to support a national kit evaluation group (KEG). Because England is within the European Union, KEG has no "licensing" function for manufacturers' kits which must be "Communautes Europeennes marked" before they can be sold within the Union. The European Union's in vitro diagnostics directive sets out common technical specifications which manufacturers must meet. There are also UK ethical constraints on the use of patient/donor blood or tissue samples which must be complied with. In this context, KEG assesses the specificity of assays in collaboration with the blood center donation testing departments. The sensitivity of assays is determined in collaboration with the Health Protection Agency and the NBS National Transfusion Microbiology Reference Laboratory using performance panels, seroconversion panels, and a large range of divergent strains to assess detection range. PMID- 17397765 TI - Jon Van Rood: pioneer at the crossroad of human leukocyte antigens and transplantation. AB - Jon Van Rood (born in 1926) has made major contributions to the fields of transfusion medicine as well as organ and stem cell transplantation. His group was the first to start unraveling the complexity of the human leukocyte antigen (HLA) system through collaborative studies that used panels of sera and leukocyte samples. Furthermore, using HLA typing, he introduced the first HLA-matched platelet transfusions and developed routine leukocyte depletion as a means to prevent HLA alloimmunization. Van Rood has also been active in the fields of kidney transplantation (Eurotransplant) and stem cell transplantation (Europdonor). He combined scientific laboratory research with application to clinical medicine. He retired from his university position in 1991 but remains active in the field. PMID- 17397766 TI - Guidelines for the use of intravenous immune globulin for hematologic and neurologic conditions. PMID- 17397767 TI - Evidence-based guidelines on the use of intravenous immune globulin for hematologic and neurologic conditions. AB - In Canada, intravenous immune globulin (IVIG) use has increased by 115% over the past 7 to 8 years. Given this increased usage, Canadian Blood Services and the National Advisory Committee on Blood and Blood Products for Canada identified the need to develop and disseminate evidence-based guidelines to facilitate appropriate IVIG use. As a result, guidelines for IVIG use in hematologic and neurologic conditions have been developed and are published in this supplement of Transfusion Medicine Reviews. This commentary provides a brief description of the process used to develop these guidelines and includes a summary of the recommendations for IVIG use in the various conditions evaluated. PMID- 17397768 TI - Guidelines on the use of intravenous immune globulin for neurologic conditions. AB - Canada's per capita use of intravenous immune globulin (IVIG) grew by approximately 115% between 1998 and 2006, making Canada one of the world's highest per capita users of IVIG. It is believed that most of this growth is attributable to off-label usage. To help ensure IVIG use is in keeping with an evidence-based approach to the practice of medicine, the National Advisory Committee on Blood and Blood Products (NAC) and Canadian Blood Services convened a panel of national experts to develop an evidence-based practice guideline on the use of IVIG for neurologic conditions. The mandate of the expert panel was to review evidence regarding use of IVIG for 22 neurologic conditions and formulate recommendations on IVIG use for each. A panel of 6 clinical experts, one expert in practice guideline development and 4 representatives from the NAC met to review the evidence and reach consensus on the recommendations for the use of IVIG. The primary sources used by the panel were 2 recent evidence-based reviews. Recommendations were based on interpretation of the available evidence and, where evidence was lacking, consensus of expert clinical opinion. A draft of the practice guideline was circulated to neurologists in Canada for feedback. The results of this process were reviewed by the expert panel, and modifications to the draft guideline were made where appropriate. This practice guideline will provide the NAC with a basis for making recommendations to provincial and territorial health ministries regarding IVIG use management. Recommendations for use of IVIG were made for 14 conditions, including acute disseminated encephalomyelitis, chronic inflammatory demyelinating polyneuropathy, dermatomyositis, diabetic neuropathy, Guillain-Barre syndrome, Lambert-Eaton myasthenic syndrome, multifocal motor neuropathy, multiple sclerosis, myasthenia gravis, opsoclonus-myoclonus, pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections, polymyositis, Rasmussen's encephalitis, and stiff person syndrome; IVIG was not recommended for 8 conditions including adrenoleukodystrophy, amyotropic lateral sclerosis, autism, critical illness polyneuropathy, inclusion body, myositis, intractable childhood epilepsy, paraproteinemic neuropathy (IgM variant), and POEMS syndrome. Development and dissemination of evidence-based clinical practice guidelines may help to facilitate appropriate use of IVIG. PMID- 17397769 TI - Guidelines on the use of intravenous immune globulin for hematologic conditions. AB - Canada's per capita use of intravenous immune globulin (IVIG) grew by approximately 115% between 1998 and 2006, making Canada one of the world's highest per capita users of IVIG. It is believed that most of this growth is attributable to off-label usage. To help ensure IVIG use is in keeping with an evidence-based approach to the practice of medicine, the National Advisory Committee on Blood and Blood Products of Canada (NAC) and Canadian Blood Services convened a panel of national experts to develop an evidence-based practice guideline on the use of IVIG for hematologic conditions. The mandate of the expert panel was to review evidence regarding use of IVIG for 18 hematologic conditions and formulate recommendations on IVIG use for each. A panel of 13 clinical experts and 1 expert in practice guideline development met to review the evidence and reach consensus on the recommendations for the use of IVIG. The primary sources used by the panel were 3 recent evidence-based reviews. Recommendations were based on interpretation of the available evidence and where evidence was lacking, consensus of expert clinical opinion. A draft of the practice guideline was circulated to hematologists in Canada for feedback. The results of this process were reviewed by the expert panel, and modifications to the draft guideline were made where appropriate. This practice guideline will provide the NAC with a basis for making recommendations to provincial and territorial health ministries regarding IVIG use management. Specific recommendations for routine use of IVIG were made for 7 conditions including acquired red cell aplasia; acquired hypogammaglobulinemia (secondary to malignancy); fetal-neonatal alloimmune thrombocytopenia; hemolytic disease of the newborn; HIV-associated thrombocytopenia; idiopathic thrombocytopenic purpura; and posttransfusion purpura. Intravenous immune globulin was not recommended for use, except under certain life-threatening circumstances, for 8 conditions including acquired hemophilia; acquired von Willebrand disease; autoimmune hemolytic anemia; autoimmune neutropenia; hemolytic transfusion reaction; hemolytic transfusion reaction associated with sickle cell disease; hemolytic uremic syndrome/thrombotic thrombocytopenic purpura; and viral-associated hemophagocytic syndrome. Intravenous immune globulin was not recommended for 2 conditions (aplastic anemia and hematopoietic stem cell transplantation) and was contraindicated for 1 condition (heparin-induced thrombocytopenia). For most hematologic conditions reviewed by the expert panel, routine use of IVIG was not recommended. Development and dissemination of evidence-based guidelines may help to facilitate appropriate use of IVIG. PMID- 17397772 TI - Defining and measuring endoscopic complications: more questions than answers. AB - The demand for information on quality in health care has risen sharply over the past decade. Endoscopic outcomes, including complication rates, need to be accurately measured and reported. Such documentation continues to be problematic because of the lack of a widely accepted classification system for endoscopic complications. Such a system should (1) include unequivocal definitions for the various types of negative outcomes and categories of complications; (2) define what negative outcomes are to be classified as complications, and (3) standardize the stratification of complications by severity. Establishing such a standardized classification of endoscopic complications could facilitate clinical research, improve the position of gastroenterologists vis-a-vis pay-for-performance programs, and result in better and more meaningful quality improvement programs, ultimately improving the care provided to patients. PMID- 17397773 TI - Minimizing complications: sedation and monitoring. AB - Serious adverse events are fortunately quite rare for procedural sedation. Current physiologic monitoring recommendations are therefore either based on "softer" outcomes, such as transient hypoxemia, or on expert opinion. Pulse oximetry and supplemental oxygen are recommended for the reduction of hypoxemia. Outcomes-based data for extended monitoring are just starting to emerge, and one of these technologies may become a recommended component of patient monitoring. With data on more than 150,000 patients published in the literature, propofol is the most studied sedative agent for gastrointestinal endoscopy. In this author's opinion, its safety and efficacy have been established. PMID- 17397774 TI - Peri-procedure pharmacotherapy, preparation, and infection control. AB - Minimizing endoscopic complications includes attention to the details about each patient's current medical condition before they undergo an invasive procedure. This article focuses on critical issues about proper preparation for endoscopic procedures and examines potential modifications to a patient's medical regimen near the time of an endoscopic procedure, highlighting recently published standards and guidelines. This article also reviews important issues related to infection control in endoscopy. PMID- 17397775 TI - Minimizing endoscopic complications--gastric and esophageal physiologic testing. AB - This article focuses on minimizing endoscopic complications associated with esophageal physiologic testing. A historical background is provided. The article then details traditional evaluation of the esophagus and stomach, as well as Bravo pH testing. PMID- 17397776 TI - Minimization of risks of esophageal dilation. AB - Esophageal dilation, along with many other therapeutic endoscopic procedures, carries with it an increased risk of complications, most of which are transient events but some of which may be catastrophic. The author reviews the literature concerning the major and minor complications that one will encounter in a busy and aggressive gastrointestinal endoscopic practice. There are scant prospective data concerning risk minimization and most studies are retrospective case series. Careful identification of groups of patients who have risk factors that may increase the potential for an adverse event is worthwhile and is reviewed. The current literature is discussed and suggestions to provide guidance and pearls in helping to avoiding and diagnosing major complications are given. PMID- 17397777 TI - Endoluminal therapy for Barrett's esophagus. AB - This article reviews methods to minimize the complications associated with endoscopic therapy for patients with Barrett's esophagus. To place this discussion in context, the natural history of Barrett's esophagus and the risks associated with progression to dysplasia and invasive carcinoma are reviewed. Operative esophageal resection traditionally is recommended for patients with Barrett's high-grade dysplasia and early carcinoma, and these surgical risks also are reviewed. Finally, all currently approved and commercially available methods for endoscopic ablation and resection of Barrett's disease are categorized according to their application methods of ablation: focal ablation, field ablation, and mucosal resection. The clinical experience with these devices is reviewed with their associated adverse events and complications. Caveats, concerns, and recommendations are discussed to help minimize the complications associated with the use of these important technologies that hold the promise of removing or destroying Barrett's disease to prevent the development of invasive carcinoma. PMID- 17397778 TI - Minimizing endoscopic complications: endoluminal stents. AB - Almost all endoluminal stents used by gastroenterologists in the United States are self-expandable metal stents, and they are placed most commonly for relief of malignant luminal obstruction. Recently, a plastic self-expandable stent was approved for treating refractory benign esophageal strictures. Endoluminal stent placement is associated with myriad of complications. Some of these complications can be avoided or minimized by the endoscopist, whereas others are beyond the endoscopist's control. This article covers the various complications that can occur following self-expandable stent placement and provides recommendations on how to minimize these complications. PMID- 17397779 TI - Minimizing complications in endoscopic retrograde cholangiopancreatography and sphincterotomy. AB - Endoscopic retrograde cholangiopancreatography (ERCP) is a major tool in the diagnosis and management of numerous biliary and pancreatic conditions, including choledocholithiasis as well as benign and malignant pancreatic diseases, especially those causing biliary obstruction. Since the procedure's inception, the techniques and indications have evolved along with advances in technology and an improved understanding of risks associated with ERCP. The trend has been away from purely diagnostic procedures; most ERCPs are now therapeutic in intent. ERCP remains among the more invasive of endoscopic procedures, with significant rates of complications that can be major. As advances are made in less invasive technology, it is important to understand the complications of ERCP and how best to avoid them. PMID- 17397780 TI - Minimizing complications of endoscopic ultrasound and EUS-guided fine needle aspiration. AB - Endoscopic ultrasound (EUS) and EUS-guided fine needle aspiration have become important tools in evaluation of patients who have various gastrointestinal and nongastrointestinal disorders, and are being increasingly utilized at many centers. With over 10 years of worldwide published clinical data, the collective experience suggests that EUS is a safe tool. There are various measures that may be undertaken to help minimize the risks. In light of ongoing advances in interventional techniques and recognition of new procedural indications, the safety of all types of procedures and efforts to minimize EUS-related complications need to be periodically reexamined. PMID- 17397781 TI - Minimizing endoscopic complications: colonoscopic polypectomy. AB - Current polypectomy tools and techniques are inadequate to prevent all postpolypectomy bleeding, perforation, and postpolypectomy syndrome; however, adherence to certain principles can substantially reduce the risk of these complications. This review does not focus on technical aspects of colonoscopy that are directed toward preventing complications of failed eradication. Rather, the authors focus on the classic complications of bleeding, perforation, and their prevention. New technologies that could further reduce or eliminate perforation and bleeding after polypectomy are sorely needed. PMID- 17397782 TI - Preventing complications of endoscopic hemostasis in acute upper gastrointestinal hemorrhage. AB - Over the last 20 years, endoscopic hemostasis has become the primary method of controlling acute, overt upper gastrointestinal hemorrhage (UGIH) and prevent rebleeding. Although uncommon, procedure-related complications may occur. This article reviews the risks attributable to endoscopic hemostasis in acute UGIH and how to help prevent these complications. PMID- 17397783 TI - Complications of capsule endoscopy. AB - Capsule endoscopy is generally a safe procedure with few complications. Clinically significant complications including symptomatic capsule retention and aspiration occur in less than 2% of examinations. There is currently no accepted means of accurately predicting or preventing capsule retention. A modified capsule has recently been developed to determine patency in patients who have possible strictures. Incomplete small bowel examinations occur in 15% of examinations. After risk stratification, various pharmacologic and endoscopic adjuncts can be employed to minimize this complication. Technical complications occur most commonly early in a center's experience and decrease as experience with the device increases. Implanted cardiac devices are no longer an absolute contraindication to the examination but should be conducted after multidisciplinary consultation. PMID- 17397784 TI - Minimizing endoscopic complications in enteral access. AB - Percutaneous enteral access techniques are important tools in the armamentarium of the skilled endoscopist. Endoscopic and post-procedural complications of enteral access are not uncommon, and the increasing population of patients requiring long-term feeding tubes places even more emphasis on minimizing them. Most enteral feeding tube complications are minor, but several have the potential to cause significant morbidity and even mortality if not recognized and managed correctly. When complications do arise, early recognition and aggressive management are essential to optimize outcomes. Expertise with the proper patient selection, choice of feeding tube and insertion techniques are critical to minimizing endoscopic complications of percutaneous enteral feeding tubes. PMID- 17397785 TI - Avoiding medicolegal complications. AB - Endoscopy can cause complications, even in the best of hands. Lawsuits are rare but more likely to be pursued when there is poor communication with patients and family before or after an event occurs. The likelihood of a suit (and of its success) can be minimized by practicing within accepted standards; by careful objective documentation of the education and consent process, and of the events themselves; and by a systematized approach to event management. PMID- 17397786 TI - Using databases to minimize endoscopic complications. AB - Providing quality clinical care as an endoscopist includes minimizing endoscopic complications. Over time, endoscopists learn which complications related to a procedure are to be expected. By using an endoscopy database, expected and unexpected complication data may be collected and assessed. This article describes what a database is, what information it contains, and how it might be used to minimize complications. In addition, the difficulties in ascertaining complications (describing them accurately, reporting them, and when possible, altering endoscopic practice to minimize them) are discussed. PMID- 17397788 TI - Molecular epidemiology of Plasmodium vivax in the State of Amazonas, Brazil. AB - Over the past 2 decades, the Amazon region of Brazil has experienced reemergence of Plasmodium vivax malaria, with reported occurrence of severe disease. The frequency and manifestations of this severe disease are unlike previous clinical experience. The hypothesis has been raised that the occurrence of severe disease may relate to the emergence of a variant form of the parasite. To test this hypothesis, we conducted a retrospective cohort study of P. vivax strains in the State of Amazonas. We determined nucleic acid sequences of segments of three genes, the 18S SSUrRNA Type A gene, the circumsporozoite surface protein (CSP) gene and the MSP-1 gene. Sequences were determined for parasites infecting 11 hospitalized (Inpatients) and 21 non-hospitalized (Outpatients) patients. We observed two common polymorphisms in the 18S SSUrRNA Type A gene; a thymidine (T)/adenine (A) polymorphism at residue 117 was significantly more common in the Inpatient group (p<0.05). Types of variation in the CSP gene included the numbers of repeat nonapeptide segments, alanine/aspartic acid polymorphism at position 5 of the nonapeptide repeat, and sporadic mutations. Alanine was more common as the fifth residue of the nonapeptide repeat in Inpatients and in strains causing second infections (both, p<0.05). Synonymous substitutions of the common repeat sequence occurred frequently in codons 1, 2, and 7, while the mutations at codon 5 were always non-synonymous, indicating that variation at codon 5 reflected selective pressure. Among MSP-1 gene sequences, recombination among progenitor strains, related to the Salvador I and Belem strains, was the main source of diversity. Phylogenetic analyses that incorporated sequence data for all three genes tested did not reveal clustering of sequences from inpatients. Our data do not affirm that the hypothesis that severe P. vivax disease in Amazonas is related to emergence of a new variant, but do suggest that variation in the fifth position of the CSP gene nonapeptide repeat may relate to disease manifestations. PMID- 17397789 TI - Sylvatic Triatoma infestans (Reduviidae, Triatominae) in the Andean valleys of Bolivia. AB - Triatoma infestans is the main vector of Chagas disease in the Southern Cone countries. Wild populations of T. infestans appear widespread throughout the Andean valleys of Bolivia. In Cotapachi (2750 m asl), all sorts of rocky outcrops, regardless of their size, provided good refuges for T. infestans. Of the 1120 ecotopes investigated, 330 (29.5%) contained triatomines and 92% of the collected insects were nymphal instars. In the cold season, triatomine densities were similar in small and large outcrops. During the hot season, bug densities were higher in the larger outcrops, particularly in those located in peridomestic sites. T. infestans populations apparently produced one generation per year. Over half the sampled bugs were positive for T. cruzi infection. At Mataral (1750 m asl), a site located in the inter-Andean Chaco, a new morph of T. infestans was detected in a sylvatic environment. PMID- 17397790 TI - Severe malaria in Cameroonian children: correlation between plasma levels of three soluble inducible adhesion molecules and TNF-alpha. AB - Plasma levels of three soluble inducible adhesion molecules, namely: intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1) and endothelial leucocyte adhesion molecule-1 (sELAM-1) or sE-selectin and the pro-inflammatory cytokine, tumour necrosis factor-alpha (TNF-alpha) were measured in well-defined clinical groups of children with severe and uncomplicated malaria. The goal of the study was to investigate the role of these molecules in immunopathogenic processes associated with severe malaria in Cameroonian children. Results showed significantly increased plasma concentrations of sICAM-1, sVCAM-1 and sE-selectin in children with severe malaria compared to those with uncomplicated malaria and healthy children (P<0.001). TNF-alpha levels increased significantly in children with severe malaria, approximately 2-folds compared to those with uncomplicated malaria and about 3-folds compared to healthy children (P<0.001). More importantly, levels of TNF-alpha strongly correlated with those of the three adhesion molecules and were significantly associated with increased risk of death (P=0.03). In addition, children who died from severe malaria showed higher mean levels of all measured factors compared to those who recovered, with significant differences observed with sICAM-1 (P<0.001) and sE-selectin (P=0.002). Furthermore, children with severe malarial anemia relative to those without, showed significantly elevated levels of the three soluble molecules; and sICAM-1 was significantly associated with increased risk of severe anemia. Taken together, these results confirm the role of TNF-alpha and the three adhesion molecules in pathogenic processes associated with severe malaria in children, and suggest an association between sICAM-1 and severe malarial anemia. PMID- 17397791 TI - An overview of lipid membrane supported by colloidal particles. AB - In recent years, original hybrid assemblies composed of a particle core surrounded by a lipid shell emerged as promising entities for various biotechnological applications. Their broadened bio-potentialities, ranging from model membrane systems or biomolecule screening supports, to substance delivery reservoirs or therapeutic vectors, are furthered by their versatility of composition due to the possible wide variation in the particle nature and size, as well as in the lipid formulation. The synthesis, the characteristics, and the uses of these Lipid/Particle assemblies encountered in the literature so far are reviewed, and classified according to the spherical core size in order to highlight general trends. Moreover, several criteria are particularly discussed: i) the interactions involved between the particles and the lipids, and implicitly the assembly elaboration mechanism, ii) the most suited techniques for an accurate characterization of the entities from structural and physicochemical points of view, and iii) the remarkable properties of the solid-supported lipid membrane obtained. PMID- 17397792 TI - A high-throughput assay shows that DNase-I binds actin monomers and polymers with similar affinity. AB - Previous conflicting reports suggest that DNase-I binds F-actin with either equal or drastically different K(D) values compared to G-actin. We developed a high throughput DNase-I inhibition assay to determine the K(D) of DNase-I for F-actin. We confirmed that phalloidin-stabilized F-actin is protected from depolymerization by DNase-I and that the critical concentration at the pointed end of phalloidin-F-actin is 45.5+/-13.9 nM. We found that DNase-I inhibition by actin follows ultrasensitive mechanics. Using varying lengths of gelsolin-capped phalloidin-F-actin, we concluded that the affinities of DNase-I for G- and the pointed end subunits of F-actin are almost indistinguishable, such that DNase-I may not distinguish between G- and F-actin conformations. PMID- 17397793 TI - The development of quantum dot calibration beads and quantitative multicolor bioassays in flow cytometry and microscopy. AB - The use of fluorescence calibration beads has been the hallmark of quantitative flow cytometry. It has enabled the direct comparison of interlaboratory data as well as quality control in clinical flow cytometry. In this article, we describe a simple method for producing color-generalizable calibration beads based on streptavidin functionalized quantum dots. Based on their broad absorption spectra and relatively narrow emission, which is tunable on the basis of dot size, quantum dot calibration beads can be made for any fluorophore that matches their emission color. In an earlier publication, we characterized the spectroscopic properties of commercial streptavidin functionalized dots (Invitrogen). Here we describe the molecular assembly of these dots on biotinylated beads. The law of mass action is used to readily define the site densities of the dots on the beads. The applicability of these beads is tested against the industry standard, namely commercial fluorescein calibration beads. The utility of the calibration beads is also extended to the characterization surface densities of dot-labeled epidermal growth factor ligands as well as quantitative indicators of the binding of dot-labeled virus particles to cells. PMID- 17397794 TI - The effects of obesity on lifting performance. AB - Obesity in the workforce is a growing problem worldwide. While the implications of this trend for biomechanical loading of the musculoskeletal system seem fairly straightforward, the evidence of a clear link between low back pain (LBP) and body mass index (BMI) (calculated as whole body mass in kilograms divided by the square of stature in meters) has not been shown in the epidemiology literature addressing this topic. The approach pursued in the current study was to evaluate the lifting kinematics and ground reaction forces of a group of 12 subjects -- six with a BMI of less than 25 kg/m(2) (normal weight) and six with a BMI of greater than 30 kg/m(2) (obese). These subjects performed a series of free dynamic lifting tasks with varied levels of load (10% and 25% of capacity) and symmetry (sagittally symmetric and 45 degrees asymmetric). The results showed that BMI had a significant effect (p<0.05) on trunk kinematics with the high BMI group exhibiting higher peak transverse plane (twisting) velocity (59% higher) and acceleration (57% higher), and exhibiting higher peak sagittal plane velocity (30% higher) and acceleration (51% higher). When normalized to body weight, there were no significant differences in the ground reaction forces between the two groups. This study provides quantitative data describing lifting task performance differences between people of differing BMI levels and may help to explain why there is no conclusive epidemiological evidence of a relationship between BMI and LBP. PMID- 17397795 TI - Spare quinones in the QB cavity of crystallized photosystem II from Thermosynechococcus elongatus. AB - The recent crystallographic structure at 3.0 A resolution of PSII from Thermosynechococcus elongatus has revealed a cavity in the protein which connects the membrane phase to the binding pocket of the secondary plastoquinone Q(B). The cavity may serve as a quinone diffusion pathway. By fluorescence methods, electron transfer at the donor and acceptor sides was investigated in the same membrane-free PSII core particle preparation from T. elongatus prior to and after crystallization; PSII membrane fragments from spinach were studied as a reference. The data suggest selective enrichment of those PSII centers in the crystal that are intact with respect to O(2) evolution at the manganese-calcium complex of water oxidation and with respect to the integrity of the quinone binding site. One and more functional quinone molecules (per PSII monomer) besides of Q(A) and Q(B) were found in the crystallized PSII. We propose that the extra quinones are located in the Q(B) cavity and serve as a PSII intrinsic pool of electron acceptors. PMID- 17397796 TI - Electron spin-echo studies of spin-labelled lipid membranes and free fatty acids interacting with human serum albumin. AB - Human serum albumin (HSA) is an abundant plasma protein that transports fatty acids and also binds a wide variety of hydrophobic pharmacores. Echo-detected (ED) EPR spectra and D(2)O-electron spin echo envelope modulation (ESEEM) Fourier transform spectra of spin-labelled free fatty acids and phospholipids were used jointly to investigate the binding of stearic acid to HSA and the adsorption of the protein on dipalmitoyl phosphatidylcholine (DPPC) membranes. In membranes, torsional librations are detected in the ED-spectra, the intensity of which depends on chain position at low temperature. Water penetration into the membrane is seen in the D(2)O-ESEEM spectra, the intensity of which decreases greatly at the middle of the membrane. Both the chain librational motion and the water penetration are only little affected by adsorption of serum albumin at the DPPC membrane surface. In contrast, both the librational motion and the accessibility of the chains to water are very different in the hydrophobic fatty acid binding sites of HSA from those in membranes. Indeed, the librational motion of bound fatty acids is suppressed at low temperature, and is similar for the different chain positions, at all temperatures. Correspondingly, all segments of the bound chains are accessible to water, to rather similar extents. PMID- 17397797 TI - Rapamycin stimulates arginine influx through CAT2 transporters in human endothelial cells. AB - In endothelial cells Tumor Necrosis Factor-alpha (TNFalpha) stimulates arginine transport through the increased expression of SLC7A2/CAT2 transcripts. Here we show that also rapamycin, an inhibitor of mTOR kinase, stimulates system y(+) mediated arginine uptake in human endothelial cells derived from either saphenous (HSVECs) or umbilical veins (HUVECs). When used together with TNFalpha, rapamycin produces an additive stimulation of arginine transport in both cell models. These effects are observed also upon incubation with AICAR, a stimulator of Adenosine Monophosphate-dependent-Protein Kinase (AMPK) that produces a rapamycin independent inhibition of the mTOR pathway. Rapamycin increases the V(max) of high affinity arginine transport and causes the appearance of a low affinity component that is particularly evident if the treatment is carried out in the presence of TNFalpha. RT-qPCR studies have demonstrated that these kinetic changes correspond to the induction of both the high affinity transporter CAT2B and the low affinity isoform CAT2A. Western blot and immunocytochemical analyses indicate that, consistently, the expression of CAT2 proteins is also stimulated under the same conditions. These changes are associated with an increase of the intracellular arginine concentration but with a decrease of NO production. Thus, our data suggest that mTOR activity is associated with the repression of CAT2 expression at mRNA and protein level. PMID- 17397798 TI - TNF-alpha inhibits BMP-induced osteoblast differentiation through activating SAPK/JNK signaling. AB - The cellular mechanism by which TNF-alpha inhibits osteoblastic differentiation induced by BMPs was investigated using mouse myoblast C2C12 cells expressing functional BMP receptors and Smad signaling molecules except ALK-6. Osteoblast transformation in response to BMP-2 was morphologically suppressed by TNF-alpha. Expression of biological markers for osteoblasts including Runx2 and osteocalcin, alkaline phosphatase activity, and parathyroid hormone (PTH) responsiveness shown by PTH-induced cAMP production were readily activated by BMP-2, -4, -6, and -7. The BMP-induced osteoblastic phenotype was dose-dependently inhibited by TNF alpha. BMP-induced Smad1,5,8 phosphorylation of C2C12 cells was suppressed by TNF alpha signaling. In addition, cDNA array analysis showed an increased expression of inhibitory Smad6 by TNF-alpha. MAP kinase analysis showed that ERK1/ERK2 and SAPK/JNK phosphorylation were selectively activated by TNF-alpha regardless of the presence of BMP ligands. BMPs had no effect on expression levels of TNF type 1 and 2 receptors. Notably, inhibition of SAPK/JNK restored TNF-alpha effects on BMP-induced osteoblast differentiation demonstrated by Id-1-promoter activity as well as Runx2 and osteocalcin mRNA levels. Collectively, TNF-alpha elicits BMP induced osteogenic inhibition by suppressing BMP-Smad signaling pathway, at least in part, through SAPK/JNK activation and Smad6 upregulation. PMID- 17397799 TI - Lack of phospholipase C-delta1 induces skin inflammation. AB - Phospholipase C (PLC) is a key enzyme in phosphoinositide signaling. We previously generated PLC-delta1 knockout (KO) mice and found that these mice showed remarkable hair loss caused by abnormalities in hair follicle structures. Here we show that the skin of PLC-delta1 KO mice displays typical inflammatory phenotypes, including increased dermal cellularity, leukocyte infiltration, and expression of pro-inflammatory cytokines. In addition, exogenously expressed PLC delta1 attenuates lipopolysaccharide-induced expression of IL-1beta, a pro inflammatory cytokine, in an enzymatic activity-dependent manner. Furthermore, suppression of skin inflammation by anti-inflammatory reagents cured the epidermal hyperplasia in PLC-delta1 KO mice. Taken together, these results indicate that lack of PLC-delta1 induces skin inflammation and that the epidermal hyperplasia in PLC-delta1 KO mice is caused by skin inflammation. Our results also suggest that PLC-delta1 regulates homeostasis of the immune system in skin. PMID- 17397800 TI - Molecular architecture of leishmania EF-1alpha reveals a novel site that may modulate protein translation: a possible target for drug development. AB - Elongation factor-1alpha plays an essential role in eukaryotic protein biosynthesis. Recently, we have shown by protein structure modeling the presence of a hairpin-loop of 12 amino acids in mammalian EF-1alpha that is absent in the leishmania homologue [D. Nandan, A. Cherkasov, R. Sabouti, T. Yi, N.E. Reiner, Molecular cloning, biochemical and structural analysis of elongation factor-1 alpha from Leishmania donovani: comparison with the mammalian homologue, Biochem. Biophys. Res. Commun. 302 (2003) 646-652]. As a consequence of this deletion, an exposed region is available on the main body of leishmania EF-1alpha. Here we report the generation of an anti-EF-1alpha antibody (DN-3) which bound selectively to the exposed region of leishmania EF-1alpha, with no reactivity with human EF-1alpha. In a leishmania cell-free protein translation system, DN-3 substantially inhibited protein translation. A similar inhibitory effect was observed when a specific peptide based on the exposed region was used in the cell free protein translation assay. The application of structure-based in silico methods to identify potential ligands to target the exposed region identified a small molecule that selectively attenuated in vitro translation using leishmania extracts. Moreover, this small molecule showed selective suppressive effect on multiplication of leishmania in culture. Taken together, these findings identify a novel, exposed region in leishmania EF-1alpha that may be involved in protein synthesis and a potential site for drug targeting. PMID- 17397801 TI - RNase HII from Chlamydia pneumoniae discriminates mismatches incorporation into DNA-rN1-DNA/DNA duplexes. AB - It was reported that RNase HII from Chlamydia pneumoniae (CpRNase HII) had RNase H activity on RNA/DNA duplex. We have analyzed the cleavage specificity of CpRNase HII on DNA-rN1-DNA/DNA duplex (rN1, one ribonucleotide). Various mismatches were introduced into the DNA-rN1-DNA/DNA duplexes at or around the ribonucleotide. The mismatches of duplexes resulted in slower cleavage rates compared to the matched duplexes. Furthermore, a greater reduction in cleavage activity was observed for the mismatches located at or adjacent to the ribonucleotide. The mismatches at the same position of DNA-rN1-DNA/DNA duplexes have different impact on the cleavage rates of CpRNase HII depending on the types of mismatches. These findings may offer further insights into the physical binding and catalytic properties of CpRNase HII-substrate interaction. PMID- 17397802 TI - Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1. AB - A novel rat gene, Mipu1, encodes a 608 amino acid protein with an amino-terminal KRAB domain and 14 carboxyl-terminal C2H2 zinc finger motifs. Mipu1 is localized to the nucleus through its KRAB domain or the linker adjacent to its zinc finger region. Using the GST-Mipu1 bound to glutathione-Sepharose beads, a consensus putative DNA binding site (5'-TGTCTTATCGAA-3') was extracted from a random oligonucleotide library. EMSA and target detection assay showed that the probe containing the putative site can bind to purified GST-Mipu1 fusion protein. The oligonucleotide containing the putative site was inserted into the pGL3-promotor vector to produce a reporter construct. The expression of reporter gene was repressed by overexpression of Mipu1 in a dose-dependent manner. Mutation analysis of the consensus sequence indicated that the repression mediated by Mipu1 is sequence-dependent. These results suggest that Mipu1 is a nuclear protein, which functions as a transcriptional repressor. PMID- 17397803 TI - FK228 inhibits Hsp90 chaperone function in K562 cells via hyperacetylation of Hsp70. AB - Some pan-histone-deacetylase (HDAC) inhibitors have recently been reported to exert their anti-leukemia effect by inhibiting the activity of class IIB HDAC6, which is the deacetylase of Hsp90 and alpha-tubulin, thereby leading to hyperacetylation of Hsp90, disruption of its chaperone function and apoptosis. In this study, we compared the effect of a class I HDAC inhibitor FK228 with the pan HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) on the Hsp90 chaperone function of K562 cells. We demonstrated that, although having a weaker inhibitory effect on HDAC6, FK228 mediated a similar disruption of Hsp90 chaperone function compared to SAHA. Unlike SAHA, FK228 did not mediate hyperacetylation of Hsp90, instead the acetylation of Hsp70 was increased and Bcr-Abl was increasingly associated with Hsp70 rather than Hsp90, forming an unstable complex that promotes Bcr-Abl degradation. These results indicated that FK228 may disrupt the function of Hsp90 indirectly through acetylation of Hsp70 and inhibition of its function. PMID- 17397804 TI - Evolution of mitochondrial cell death pathway: Proapoptotic role of HtrA2/Omi in Drosophila. AB - Despite the essential role of mitochondria in a variety of mammalian cell death processes, the involvement of mitochondrial pathway in Drosophila cell death has remained unclear. To address this, we cloned and characterized DmHtrA2, a Drosophila homolog of a mitochondrial serine protease HtrA2/Omi. We show that DmHtrA2 normally resides in mitochondria and is up-regulated by UV-irradiation. Upon receipt of apoptotic stimuli, DmHtrA2 is translocated to extramitochondrial compartment; however, unlike its mammalian counterpart, the extramitochondrial DmHtrA2 does not diffuse throughout the cytosol but stays near the mitochondria. RNAi-mediated knock-down of DmHtrA2 in larvae or adult flies results in a resistance to stress stimuli. DmHtrA2 specifically cleaves Drosophila inhibitor of-apoptosis protein 1 (DIAP1), a cellular caspase inhibitor, and induces cell death both in vitro and in vivo as potent as other fly cell death proteins. Our observations suggest that DmHtrA2 promotes cell death through a cleavage of DIAP1 in the vicinity of mitochondria, which may represent a prototype of mitochondrial cell death pathway in evolution. PMID- 17397805 TI - Perineuronal nets protect against amyloid beta-protein neurotoxicity in cultured cortical neurons. AB - Perineuronal nets (PNs) consisting of chondroitin sulfate proteoglycans (CSPGs) and hyaluronic acid are associated with distinct neuronal populations in mammalian brain. Cortical areas abundant in PNs have been known to be less affected by neurotoxicity in human Alzheimer's disease. In the present study, we examined whether PNs protect the neurotoxicity caused by amyloid beta-protein (Abeta), a major constituent of senile plaques in Alzheimer's disease using cortical neurons of dissociated culture. Double labeling experiments using confocal microscopy showed that the neurons associated with PNs were visualized with the anti-CSPG antibody in dissociated cortical culture. The analysis of reverse transcription-polymerase chain reaction revealed that mRNA expression of chondroitin sulfotransferases, CSPG-specific enzymes, was detected in neuronal culture, indicating that cultured cortical neurons are able to synthesize CSPGs and construct PNs structure. The treatment of Abeta1-42 showed significant neurotoxicity on PNs-free cortical neurons, however, it did not reveal neurotoxicity on PNs-associated neurons. Moreover, it was shown that the treatment of Abeta1-42 was able to kill PNs-associated neurons after the removal of chondroitin sulfate (CS) glycosaminoglycans with chondroitinase ABC. The treatment of glutamate killed not only PNs-free cortical neurons but also PNs associated neurons. These results suggest that CS glycosaminoglycans on PNs are responsible for protecting neurons from Abeta1-42 neurotoxicity. PMID- 17397806 TI - Status epilepticus induces region-specific changes in dendritic spines, dendritic length and TrkB protein content of rat brain cortex. AB - Induction of status epilepticus (SE) with kainic acid results in a large reorganization of neuronal brain circuits, a phenomenon that has been studied primarily in the hippocampus. The neurotrophin BDNF, by acting through its receptor TrkB, has been implicated in such reorganization. In the present work we investigated, by Western blot and immunohistochemistry, whether regional changes of TrkB expression within the rat brain cortex are correlated with altered neuronal morphology and/or with apoptotic cell death. We found that the full length TrkB protein decreased within the cortex when measured 24 h to 1 week after induction of SE. Analysis by immunohistochemistry revealed that TrkB staining diminished within layer V of the retrosplenial granular b (RSGb) and motor cortices, but not within the auditory cortex. In layer II/III, differential changes were also observed: TrkB decreased in the motor cortex, did not change within the RSGb but increased within the auditory cortex. Reduced TrkB was associated with dendritic atrophy and decreased spine density in pyramidal neurons within layer V of the RSGb. No correlation was observed between regional and cellular changes of TrkB protein and apoptosis, measured by the TdT-mediated dUTP nick end labeling (TUNEL) method. The global decrease of TrkB within the neocortex and the associated dendritic atrophy may counteract seizure propagation in the epileptic brain but may also underlie cognitive impairment after seizures. PMID- 17397807 TI - Activity of attention related structures in multiple sclerosis patients. AB - AIMS: It was investigated whether the cortical activity during tasks requiring focused or divided attention is reduced in multiple sclerosis patients with prominent deficits (MS(+D)) and increased in patients without impairment (MS(-D)) in these specific attention functions. METHODS: Six MS(+D) patients with attention deficits, six unimpaired MS(-D) patients, and age-, gender-, and education-matched healthy control subjects were examined with functional magnetic resonance imaging (fMRI). The experimental paradigm consisted of visual tasks requiring focused or divided attention. RESULTS: Performance accuracy and reaction times were impaired in MS(+D) patients. This subgroup showed reduced activation within superior and inferior frontal gyrus during focused attention. Under conditions of divided attention decreased activity was found within middle and inferior frontal gyrus, inferior parietal structures, and occipital areas. No compensatory activity was observed. MS(-D) patients did neither differ in behavioral data nor in cortical activity in attention related structures from control subjects. CONCLUSIONS: The study found evidence for the neural correlate of attentional deficits in MS patients. In patients with specific attention deficits, reduced cortical activity in prefrontal and parietal areas, which are associated with attention and executive control, reflects the patients' reduced performance on a behavioral level. Our findings also suggest impaired top-down attentional control on sensory structures in these patients. In patients without verifiable attention deficits a normal functioning of structures relevant for executive attention was observed. Compensatory activity in these structures as a marker of reorganization in less pronounced stages of the disease was not found. PMID- 17397808 TI - Cholinergic projections to the suprachiasmatic nucleus and lower subparaventricular zone of diurnal and nocturnal rodents. AB - In nocturnal species cholinergic agonists alter circadian rhythm phase when injected intraventricularly or directly into the suprachiasmatic nucleus (SCN), but the phase shifts obtained differ depending upon the site being injected. Cholinergic projections reach the SCN of nocturnal laboratory rats, however, nothing is known about these projections in diurnal rodents. The first objective of this study was to evaluate the hypothesis that cholinergic projections to the SCN are only present in nocturnal species. The second objective was to evaluate the hypothesis that the lower part of the subparaventricular zone (LSPV) is a candidate for being a site that mediates the phase shifts observed when cholinergic agonists are injected intraventricularly. These hypotheses were tested in the diurnal unstriped Nile grass rat (Arvicanthis niloticus) and the nocturnal laboratory rat. Additionally, we evaluated if the light-dark (LD) cycle had an effect on the expression of the vesicular acetylcholine transporter (VAChT) in the SCN, LSPV, and in two control areas. Animals were kept in a 12:12 LD cycle and perfused at six time points. VAChT immunoreactivity was observed in the SCN, LSPV, and in the control areas of both species. The SCN and LSPV showed a differential distribution and density of cholinergic projections between the two species, but similar temporal patterns of VAChT expression were found across species. These results provide evidence for a differential cholinergic stimulation of the SCN between grass rats and laboratory rats that may reflect a rewiring of neural projections brought about by the adoption of a diurnal activity profile. PMID- 17397809 TI - Seizure activity affects neuroglial Kv1 channel immunoreactivities in the gerbil hippocampus. AB - In order to confirm the species-specific distribution of voltage-gated K(+) (Kv) channels and the definitive relationship between their immunoreactivities and seizure activity, we investigated Kv1 channel immunoreactivities in the hippocampus of seizure resistant (SR) and seizure sensitive (SS) gerbils. There was distinct difference of the Kv1 channel subtypes immunoreactivity in the hippocampi in both SR and SS gerbils. Kv1.1, Kv1.2, Kv1.3, Kv1.4, and Kv1.6 immunoreactivities in the SS gerbil hippocampus were lower than that in the SR gerbil hippocampus. However, Kv1 immunoreactivities were obviously presented in astrocyte within the stratum radiatum of the CA1 region of pre-seizure SS gerbil hippocampus. Following seizure-onset, Kv1 immunoreactivities (except Kv1.5) were markedly elevated, whereas their immunoreactivites in astrocytes were down regulated. Therefore, the present study demonstrates that seizure activity may distinctly affect neuroglial Kv1 immunoreactivities in the gerbil hippocampus. PMID- 17397810 TI - Rapid and long-term induction of effector immediate early genes (BDNF, Neuritin and Arc) in peri-infarct cortex and dentate gyrus after ischemic injury in rat brain. AB - The genomic response following brain ischemia is very complex and involves activation of both protective and detrimental signaling pathways. Immediate early genes (IEGs) represent the first wave of gene expression following ischemia and are induced in extensive regions of the ischemic brain including cerebral cortex and hippocampus. Brain-derived neurotrophic factor (BDNF), Neuritin and Activity regulated cytoskeleton-associated protein (Arc) belong to a subgroup of immediate early genes implicated in synaptic plasticity known as effector immediate early genes. Here, we investigated the spatial and temporal activation pattern for these genes during the first 24 h of reperfusion following 2-h occlusion of the middle cerebral artery. Neuritin showed a persistent activation in frontal cingulate cortex while Arc displayed a biphasic response. Also, in dentate gyrus, activation was observed at 0-6 h of reperfusion for Neuritin and 0-12 h of reperfusion for Arc while BDNF was induced 0-9 h of reperfusion. Our study demonstrates a rapid and long-term activation of effector immediate early genes in distinct brain areas following ischemic injury in rat. Effector gene activation may be part of long-term synaptic responses of ischemic brain tissue. PMID- 17397811 TI - Effects of acetylpuerarin on hippocampal neurons and intracellular free calcium subjected to oxygen-glucose deprivation/reperfusion in primary culture. AB - OBJECTIVES: This study was undertaken to find out the effects of acetylpuerarin on hippocampal neurons and intracellular free calcium in primary culture subjected to oxygen-glucose deprivation/reperfusion. METHODS: According to different reperfusion time (1 h, 6 h, 12 h, 24 h), three concentrations (1.6 micromol l(-1), 0.4 micromol l(-1), 0.1 micromol l(-1)) of acetylpuerarin, and MK 801 (10 micromol l(-1)), a positive control drug, neurons were randomly divided into 21 groups. Each group was observed by inverted phase contrast microscope; neuron viability was measured by the reduction of 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT); intracellular Ca(2+) was observed by Fura 2/AM ester through fluorospectrophotometer. RESULTS: The injured neurons were protected and degeneration and necrosis were alleviated in treatment groups of acetylpuerarin and MK-801. Acetylpuerarin increased the neuron viability at high, middle and low concentrations. Fluorescence detection results showed that the calcium concentration in the group treated with acetylpuerarin and MK-801 was lowered in each reperfusion time. CONCLUSION: Our results demonstrated that acetylpuerarin could protect the hippocampal neurons from ischemia-reperfusion injury in rats by alleviating the morphological damage, increasing neuron viability and decreasing calcium concentration in neuron. PMID- 17397812 TI - Fluoro-Jade C can specifically stain the degenerative neurons in the substantia nigra of the 1-methyl-4-phenyl-1,2,3,6-tetrahydro pyridine-treated C57BL/6 mice. AB - Fluoro-Jade C, a new-developed fluorescent dye, has been successfully applied for identification of neuronal degeneration in the substantia nigra of 1-methyl-4 phenyl-1,2,3,6-tetrahydro pyridine (MPTP)-treated mice in the present study. The animal model was first prepared by intraperitoneal injection of neurotoxicant MPTP that can specifically induce degeneration of dopamine neurons in the substantia nigra of C57BL/6 mice. Fluoro-Jade C was then utilized to stain the midbrain sections and semiquantitation analysis was carried out in comparison with controls. It revealed that Fluoro-Jade C-positive cells showed strong green color in neuronal profile and were observed in the substantia nigra of MPTP treated mice whereas they were not detected in that of controls. The Fluoro-Jade C-positive cells were mostly shrunken or smaller-sized in their cell bodies in comparing with that of normal dopamine neurons of controls. In the midbrain of MPTP-treated mice, Fluoro-Jade C-positive neuronal cells were exclusively distributed in the substantia nigra pars compacta, but rarely seen in the ventral tegemental area where dopamine neurons were numerously distributed. Double labeling experiments indicated that a population of Fluoro-Jade C-positive cells (23%) exhibited neuron-specific nuclear protein-immunoreactivity and none of them showed immunoreactivity to glial cell marker glial fibrillary acid protein. However, most of Fluoro-Jade C-positive degenerative neurons (98%) lost their immunoreactivity to dopaminergic marker tyrosine hydroxylase in the substantia nigra of MPTP-treated mice. Taken together with previous observations, this study has presented that Fluoro-Jade C can be sensitively and specifically utilized to identify the neuronal degeneration in the substantia nigra of rodent animals receiving MPTP insult. PMID- 17397813 TI - Dysequilibrium between caspases and their inhibitors in a mouse model for amyotrophic lateral sclerosis. AB - Mutations in copper/zinc superoxide dismutase (SOD1) have been implicated in the pathogenesis of familial amyotrophic lateral sclerosis (ALS). Mutant SOD1 protein likely gains a novel cytotoxic property, leading to the death of motor neurons. We therefore investigated whether caspase-mediated apoptosis is associated with novel cytotoxic properties in a rodent model for familial ALS (G93A SOD1 transgenic mice). Caspase-9 (an effecter in the mitochondrial apoptotic pathway), caspase-8 (an effecter in the Fas apoptotic pathway), and caspase-3 (an executioner of both pathways) proteins were all present in nonactive forms in the spinal cords of wild-type mice during the early stage of the disease (8 weeks), at which time the mice had not yet exhibited motor paralysis. In transgenic mice, however, these proteins were present in their active forms, and their mRNA levels were significantly upregulated in the represent to this conversion from nonactive to active forms. During the advanced stage of the disease (16 weeks), when paralysis was evident, the active caspase levels were further elevated. On the other hand, the mRNA and protein levels of survivin, a counteraction protein against caspases, were significantly suppressed during the early stage, and sharply increased during the advanced stage. Although the mRNA and protein levels of X-linked inhibitor of apoptosis protein (XIAP) remained at the same levels as those seen in the control (wild-type mice) during the early stage, they were significantly depressed at an age of 16 weeks. These findings were observed exclusively in the spinal cord, the region responsible for the disease, and not in the cerebellum, a non-responsible region. We conclude that conditions facilitating the apoptotic process during the early stage of the disease play causative roles in the pathogenesis of ALS and that the suppression of XIAP levels during the advanced stage could contribute to disease expression and/or progression. PMID- 17397814 TI - Novel chalcogenides of thymidine and uridine: synthesis, properties and applications. AB - A facile and efficient methodology has been developed for the synthesis of dithymidine and di-uridine derived disulfides using benzyltriethylammonium tetrathiomolybdate as a sulfur transfer reagent. However, a similar reaction of thymidine derivative with tetraethylammonium tetraselenotungstate as a selenium transfer reagent resulted in the formation of an unexpected cyclic diselenide. The disulfide derivatives of nucleosides have been used as precursors in a tandem disulfide cleavage-Michael addition/ring opening reactions to construct aminoacid and carbocyclic derivatives of nucleosides. PMID- 17397815 TI - Development of a physiologically based toxicokinetic model for butadiene and four major metabolites in humans: global sensitivity analysis for experimental design issues. AB - 1,3-Butadiene (BD) is metabolized in humans and rodents to mutagenic and carcinogenic species. Our previous work has focused on developing a physiologically based toxicokinetic (PBTK) model for BD to estimate its metabolic rate to 1,2-epoxy-3-butene (EB), using exhaled breath BD concentrations in human volunteers exposed by inhalation. In this paper, we extend our BD model to describe the kinetics of its four major metabolites EB, 1,2:3,4-diepoxybutane (DEB), 3-butene-1,2-diol (BDD), and 3,4-epoxy-1,2-butanediol (EBD), and to test whether the extended model and experimental data (to be collected for BD and metabolites in humans) are together adequate to estimate the metabolic rate constants of each of the above chemicals. Global sensitivity analyses (GSA) were conducted to evaluate the relative importance of the model parameters on model outputs during the 20min of exposure and the 40min after exposure ended. All model parameters were studied together with various potentially measurable model outputs: concentrations of BD and EB in exhaled air, concentrations of BD and all metabolites in venous blood, and cumulated amounts of urinary metabolites excreted within 24h. Our results show that pulmonary absorption of BD and subsequent distribution and metabolism in the well-perfused tissues compartment are the critical processes in the toxicokinetics of BD and metabolites. In particular, three parameters influence numerous outputs: the blood:air partition coefficient for BD, the metabolic rate of BD to EB, and the volume of the well perfused tissues. Other influential parameters include other metabolic rates, some partition coefficients, and parameters driving the gas exchanges (in particular, for BD outputs). GSA shows that the impact of the metabolic rate of BD to EB on the BD concentrations in exhaled air is greatly increased if a few of the model's important parameters (such as the blood:air partition coefficient for BD) are measured experimentally. GSA also shows that all the transformation pathways described in the PBTK model may not be estimable if only data on the studied outputs are collected, and that data on a specific output for a chemical may not inform all the transformations involving that chemical. PMID- 17397816 TI - DNA repair pathways and mitochondrial DNA mutations in gastrointestinal carcinogenesis. AB - This work focuses on the main DNA repair pathways, highlighting their role in gastrointestinal carcinogenesis and the role of mitochondrial DNA (mtDNA), mutations being described in several tumor types, including those of the gastrointestinal tract. The mismatch repair (MMR) system is inherently altered in patients with hereditary non-polyposis colorectal cancer, and plays a role in carcinogenesis in a subset of sporadic colorectal, gastric and esophageal cancers. Alterations in homologous recombination (HR) and non-homologous end joining (NHEJ) also contribute to the development of pancreatic cancer. Gene polymorphisms of some X-ray cross-complementing (XRCCs), cofactor proteins involved in the base excision repair pathway, have been investigated in relation to gastric, colorectal and pancreatic cancer. Yet only one polymorphism, XRCC1 Arg194Trp, appears to be involved in smoking-related cancers and in early onset pancreatic cancer. Although evidence in the literature indicates that mtDNA somatic mutations play a role in gastric and colorectal carcinogenesis, no sound conclusions have yet been drawn regarding this issue in pancreatic cancer, although an mtDNA variant at 16519 is believed to worsen the outcome of pancreatic cancer patients, possibly because it is involved in altering cellular metabolism. PMID- 17397817 TI - Small Dendrobaena earthworms survive freezing better than large worms. AB - Dendrobaena octaedra is a freeze tolerant earthworm widely distributed in boreal regions. Specimens collected in Sweden were cold acclimated and then frozen at -7 degrees C to examine the influence of body mass on survival of freezing. Results showed that survival was negatively correlated to body mass. Glycogen content of the worms was variable and seemed to decrease with increasing body mass consistent with the hypothesis that freeze survival is dependent on the ability to rapidly break down glycogen and accumulate high concentrations of glucose. The results suggest that large worms (subadults and adults) invest energy in production of cocoons at the expense of glycogen storage for cryoprotectant production, whereas juvenile worms increase their survival chances by investing energy in glycogen storage at the expense of growth as a preparation for winter. PMID- 17397818 TI - Wild-type cells rescue genotypically Math1-null hair cells in the inner ears of chimeric mice. AB - The transcription factor Math1 has been shown to be critical in the formation of hair cells (HCs) in the inner ear. However, the influence of environmental factors in HC specification suggests that cell extrinsic factors are also crucial to their development. To test whether extrinsic factors impact development of Math1-null (Math1(beta-Gal/beta-Gal)) HCs, we examined neonatal (postnatal ages P0-P4.5) Math1-null chimeric mice in which genotypically mutant and wild-type cells intermingle to form the inner ear. We provide the first direct evidence that Math1-null HCs are able to be generated and survive in the conducive chimeric environment. beta-Galactosidase expression was used to identify genetically mutant cells while cells were phenotypically defined as HCs by morphological characteristics notably the expression of HC-specific markers. Genotypically mutant HCs were found in all sensory epithelia of the inner ear at all ages examined. Comparable results were obtained irrespective of the wild-type component of the chimeric mice. Thus, genotypically mutant cells retain the competence to differentiate into HCs. The implication is that the lack of the Math1 gene in HC precursors can be overcome by environmental influences, such as cell-cell interactions with wild-type cells, to ultimately result in the formation of HCs. PMID- 17397819 TI - Development of a chordate anterior-posterior axis without classical retinoic acid signaling. AB - Developmental signaling by retinoic acid (RA) is thought to be an innovation essential for the origin of the chordate body plan. The larvacean urochordate Oikopleura dioica maintains a chordate body plan throughout life, and yet its genome appears to lack genes for RA synthesis, degradation, and reception. This suggests the hypothesis that the RA-machinery was lost during larvacean evolution, and predicts that Oikopleura development has become independent of RA signaling. This prediction raises the problem that the anterior-posterior organization of a chordate body plan can be developed without the classical morphogenetic role of RA. To address this problem, we performed pharmacological treatments and analyses of developmental molecular markers to investigate whether RA acts in anterior-posterior axial patterning in Oikopleura embryos. Results revealed that RA does not cause homeotic posteriorization in Oikopleura as it does in vertebrates and cephalochordates, and showed that a chordate can develop the phylotypic body plan in the absence of the classical morphogenetic role of RA. A comparison of Oikopleura and ascidian evidence suggests that the lack of RA induced homeotic posteriorization is a shared derived feature of urochordates. We discuss possible relationships of altered roles of RA in urochordate development to genomic events, such as rupture of the Hox-cluster, in the context of a new understanding of chordate phylogeny. PMID- 17397820 TI - Transcriptional repressor and activator activities of SMA-9 contribute differentially to BMP-related signaling outputs. AB - In the nematode Caenorhabditis elegans, the BMP-related growth factor DBL-1 regulates body size and male tail morphogenesis via a conserved receptor/Smad signaling pathway. Smads are transcription factors, but rely on transcription cofactors for appropriate regulation of target genes in response to TGF-beta- and BMP-related signals. In the DBL-1 pathway, sma-9 encodes multiple zinc finger transcription factors homologous to Drosophila Schnurri, which functions in Dpp/BMP signaling. We have studied the molecular functions of SMA-9 as a model for transcription cofactor-dependent regulation of gene expression. Using SMA-9 fusions to known transcriptional activators and repressors, we demonstrate that SMA-9 acts primarily as a transcriptional repressor in body size regulation in vivo. In contrast, both activator and repressor functions contribute to male tail patterning. We further show that different SMA-9 regions have intrinsic repressor and activator activities using a yeast transcription assay. We use microarray analysis to identify transcriptional target genes in body size regulation. Consistent with the importance of repression in mediating body size regulation, we find more repressed genes than activated genes in this pool. Finally, we identify five transcriptional targets with body size and/or male tail patterning phenotypes, including transcription factors related to Runx and fos and signaling molecules related to hedgehog and patched. Our results thus suggest that SMA-9 products function differentially as transcriptional repressors and activators in DBL-1/BMP pathway regulated body size and male tail morphogenesis. PMID- 17397821 TI - Gastrulation in the cnidarian Nematostella vectensis occurs via invagination not ingression. AB - Gastrulation is a central event in metazoan development, involving many cellular behaviors including invagination, delamination, and ingression. Understanding the cell biology underlying gastrulation in many different taxa will help clarify the evolution of gastrulation mechanisms. Gastrulation in the anthozoan cnidarian Nematostella vectensis has been described as a combination of invagination and unipolar ingression through epithelial to mesenchymal transitions (EMT), possibly controlled by snail genes, important regulators of EMT in other organisms. Our examination, however, fails to reveal evidence of ingressing cells. Rather, we observe that endodermal cells constrict their apices, adopting bottle-like morphologies especially pronounced adjacent to the blastopore lip. They retain apical projections extending to the archenteron throughout gastrulation. Basally, they form actin-rich protrusions, including interdigitating filopodia that may be important in pulling the ectodermal and endodermal cells together. Endodermal cells retain cell-cell junctions while invaginating, and are organized throughout development. Never is the blastocoel filled by a mass of mesenchyme. Additionally, injection of splice-blocking morpholinos to Nematostella snail genes does not result in a phenotype despite dramatically reducing wild-type transcript, and overexpression of Snail-GFP in different clonal domains has no effect on cell behavior. These data indicate that EMT is not a major factor during gastrulation in Nematostella. PMID- 17397822 TI - The Wnt inhibitor, Dickkopf 4, is induced by canonical Wnt signaling during ectodermal appendage morphogenesis. AB - Ectodermal appendage morphogenesis requires continuous epithelial-mesenchymal cross-talk during development. Canonical Wnt signaling has been shown to be pivotal during this process and its inhibition leads to the absence of any morphological or molecular signs of appendage formation, including hair follicles (HFs). In the mouse, primary HFs arise in utero starting just before E14.5, when the first morphological signs of a placode are discernible. In this study, our goal was to identify novel factors expressed during primary HF morphogenesis. We performed transcriptional profiling of the developing epidermis at 12 h intervals between E12.5 and E15.5. One of the significantly differentially expressed genes was the Wnt inhibitor Dickkopf 4, Dkk4. We show that Dkk4 mRNA increases sharply in the dorso-lateral epidermis around E14 and then decreases until E15.5. Using whole mount in situ hybridization, we show that Dkk4 mRNA is localized to the pre placodes at sites of presumptive epithelial-mesenchymal interactions during appendage morphogenesis, including the dental lamina, mammary gland, eccrine gland, and primary and secondary HFs. In silico analysis, reporter gene assays as well as in vitro transfections of LEF1 and beta-catenin show that Dkk4 is a potential downstream target of canonical Wnt signaling. In addition, we demonstrate a direct physical interaction between LEF1/beta-catenin complex and the Dkk4 promoter using ChIP. We propose that Dkk4 acts in a negative feedback loop to attenuate canonical Wnt signaling, and may facilitate a switch to the non canonical Wnt planar cell polarity (PCP) pathway that is involved in cell movements during morphogenesis. PMID- 17397823 TI - Heat injury-induced drop of the noxious heat threshold measured with an increasing-temperature water bath: a novel rat thermal hyperalgesia model. AB - Conventional thermonociceptive tests are based on measurement of the latency of nocifensive reactions evoked by constant, suprathreshold heat stimuli. In the present study, a novel, increasing-temperature water bath was developed for determination of the noxious heat threshold temperature of lightly restrained conscious rats. One of the hindpaws of a rat was immersed into the water bath whose temperature was increased from 30 degrees C at a rate of 24 degrees C/min until the animal withdrew its hindpaw from the water. The corresponding bath temperature was considered as behavioural noxious heat threshold. The heat threshold of untreated rats was 43.5+/-0.4 degrees C (n=10) and was reproducible upon repeated measurements at intervals of 10 min for 60 min. Thermal hyperalgesia was induced by mild heat injury (51 degrees C water for 20 s) which led to a 7-8 degrees C decrease of the noxious heat threshold. Thermal hyperalgesia was detected at least for 60 min after heat injury. Morphine, diclofenac, ibuprofen and paracetamol administered intraperitoneally 20 min after heat injury dose-dependently inhibited the drop of heat threshold with minimum effective doses of 0.3, 0.3, 10 and 30 mg/kg, and ED(50) values of 0.5, 3, 18 and 100 mg/kg, respectively. Thermal hyperalgesia was also decreased by intraplantar treatment with morphine (10 microg), diclofenac (10 microg) or ibuprofen (100 microg). In conclusion, the mild heat injury-induced drop of the noxious heat threshold measured with the increasing-temperature water bath is a novel thermal hyperalgesia model highly sensitive to both opioid and non-opioid analgesics upon systemic or local administration. PMID- 17397824 TI - Poly(ADP-ribose) polymerase inhibition improves endothelial dysfunction induced by reactive oxidant hydrogen peroxide in vitro. AB - Reactive oxygen species, such as hydrogen peroxide (H(2)O(2)) induce oxidative stress and DNA-injury. The subsequent activation of poly(ADP-ribose) polymerase (PARP) has been implicated in the pathogenesis of various cardiovascular diseases including ischaemia-reperfusion injury, circulatory shock, diabetic complications and atherosclerosis. We investigated the effect of PARP-inhibition on endothelial dysfunction induced by H(2)O(2). In vascular reactivity measurements on isolated rat aortic rings we investigated the phenylephrine-induced contraction, and endothelium-dependent and -independent vasorelaxation by using cumulative concentrations of acetylcholine and sodium nitroprusside. Endothelial dysfunction was induced by exposing the rings to H(2)O(2) (200 and 400 muM) for 30 min. In the treatment group, rings were preincubated with the potent PARP-inhibitor INO 1001. DNA strand breaks were assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method. Immunohistochemical analysis was performed for poly(ADP-ribose) (the enzymatic product of PARP) and for apoptosis inducing factor (a pro-apoptotic factor regulated by PARP). Exposure to H(2)O(2) resulted in reduced contraction forces and a dose-dependent impairment of endothelium-dependent vasorelaxation of aortic rings (maximal relaxation to acetylcholine: 86.21+/-1.574% control vs. 72.55+/-1.984% H(2)O(2) 200 muM vs. 66.86+/-1.961% H(2)O(2) 400 muM; P<0.05). PARP-inhibition significantly improved the acetylcholine-induced vasorelaxation (77.75+/-3.019% vs. 66.86+/-1.961%; P<0.05), while the contractility remained unaffected. The dose-response curves of endothelium-independent vasorelaxation to sodium nitroprusside did not differ in any groups studied. In the H(2)O(2) groups immunohistochemical analysis showed enhanced PARP-activation and nuclear translocation of apoptosis inducing factor, which were prevented by INO-1001. Our results demonstrate that PARP activation contributes to the pathogenesis of H(2)O(2)-induced endothelial dysfunction, which can be prevented by PARP inhibitors. PMID- 17397825 TI - Molecular mechanism of arachidonic acid inhibition of the CFTR chloride channel. AB - Arachidonic acid inhibits the activity of a number of different Cl- channels, however its molecular mechanism of action is not known. Here we show that inhibition of cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels by arachidonic acid is weakened following mutagenesis of two positively charged pore-lining amino acids. Charge-neutralizing mutants K95Q and R303Q both increased the Kd for inhibition from approximately 3.5 microM in wild type to approximately 17 microM. At both sites, the effects of mutagenesis were dependent of the charge of the substituted side chain. We suggest that arachidonic acid interacts electrostatically with positively charged amino acid side chains in the cytoplasmic vestibule of the CFTR channel pore to block Cl- permeation. PMID- 17397826 TI - Inhibition of fatty acid amide hydrolase, a key endocannabinoid metabolizing enzyme, by analogues of ibuprofen and indomethacin. AB - There is evidence in the literature that the nonsteroidal anti-inflammatory drugs indomethacin and ibuprofen can interact with the cannabinoid system both in vitro and in vivo. In the present study, a series of analogues of ibuprofen and indomethacin have been investigated with respect to their ability to inhibit fatty acid amide hydrolase, the enzyme responsible for the hydrolysis of the endogenous cannabinoid anandamide. Of the fourteen compounds tested, the 6-methyl pyridin-2-yl analogue of ibuprofen ("ibu-am5") was selected for further study. This compound inhibited rat brain anandamide hydrolysis in a non-competitive manner, with IC50 values of 4.7 and 2.5 microM being found at pH 6 and 8, respectively. By comparison, the IC50 values for ibuprofen were 130 and 750 microM at pH 6 and 8, respectively. There was no measurable N-acylethanolamine hydrolyzing acid amidase activity in rat brain membrane preparations. In intact C6 glioma cells, ibu-am5 inhibited the hydrolysis of anandamide with an IC50 value of 1.2 microM. There was little difference in the potencies of ibu-am5 and ibuprofen towards cyclooxygenase-1 and -2 enzymes, and neither compound inhibited the activity of monoacylglycerol lipase. Ibu-am5 inhibited the binding of [3H] CP55,940 to rat brain CB1 and human CB2 cannabinoid receptors more potently than ibuprofen, but the increase in potency was less than the corresponding increase in potency seen for inhibition of FAAH activity. It is concluded that ibu-am5 is an analogue of ibuprofen with a greater potency towards fatty acid amide hydrolase but with a similar cyclooxygenase inhibitory profile, and may be useful for the study of the therapeutic potential of combined fatty acid amide hydrolase cyclooxygenase inhibitors. PMID- 17397827 TI - Adenoviral-mediated expression of dihydropyridine-insensitive L-type calcium channels in cardiac ventricular myocytes and fibroblasts. AB - Cardiac voltage-gated Ca2+ channels regulate the intracellular Ca2+ concentration and are therefore essential for muscle contraction, second messenger activation, gene expression and electrical signaling. As a first step in accessing the structural versus functional properties of the L-type Ca2+ channel in the heart, we have expressed a dihydropyridine (DHP)-insensitive CaV1.2 channel in rat ventricular myocytes and fibroblasts. Following isolation and culture, cells were infected with adenovirus expressing either LacZ or a mutant CaV1.2 channel (CaV1.2DHPi) containing the double mutation (T1039Y & Q1043M). This mutation renders the channel insensitive to neutral DHP compounds such as nisoldipine. The whole-cell, L-type Ca2+ current (ICa) measured in control myocytes was inhibited in a concentration-dependent manner by nisoldipine with an IC50 of 66 nM and complete block at 250 nM. In contrast, ICa in cells infected with AdCaV1.2DHPi was inhibited by only 35% by 500 nM nisoldipine but completely blocked by 50 microM diltiazem. In order to study CaV1.2DHPi in isolation, myocytes infected with AdCaV1.2DHPi were incubated with nisoldipine. Under this condition the cells expressed a large ICa (12 pA/pF) and displayed Ca2+ transients during field stimulation. Furthermore, addition of 2 microM forskolin and 100 microM 3 isobutyl-1-methylxanthine (IBMX), to stimulate protein kinase A, strongly increased IBa in the AdCaV1.2DHPi-infected cells. A Cd2+-sensitive IBa was also recorded in cardiac fibroblasts infected with AdCaV1.2DHPi. Thus, expression of CaV1.2DHPi will provide an important tool in studies of cardiac myocyte and fibroblast function. PMID- 17397828 TI - Neuroprotective effects of the novel polyethylene glycol-hemoglobin conjugate SB1 on experimental cerebral thromboembolism in rats. AB - SunBio1 (SB1) is a novel polyethylene glycol-bovine hemoglobin conjugate. It is a small molecule that shows high oxygen-delivery capacity, and exhibits extended plasma half-life compared to hemoglobin alone, thus reducing renal toxicity. The aim of the present study was to evaluate potential neuroprotective effects of SB1 using a rat middle cerebral artery occlusion model. The middle cerebral artery of male Sprague-Dawley rats was occluded with a thrombotic blood clot and SB1 was administered via intra-arterial infusion 5 min after the operation. Brain tissue was harvested after 2 h, and cerebral infarct volumes were calculated from coronal sections stained with 2,3,5-triphenyltetrazolium chloride. Three to 6 days after the procedure, sub-groups of animals were subjected to an open field test and the Morris water maze to assess locomotor activity and learning/memory function. Thrombotic blood clots induced extensive brain infarction and edema; however, these were significantly reduced in SB1 treated animals. In addition, SB1 treatment increased locomotor activity in open field tests, and improved the learning/memory deficits caused by the thromboembolism. These results suggest that SB1 has neuroprotective effects against ischemic brain injury caused by thromboembolism. PMID- 17397829 TI - Recombinant human ciliary neurotrophic factor reduces weight partly by regulating nuclear respiratory factor 1 and mitochondrial transcription factor A. AB - Ciliary neurotrophic factor (CNTF) can lead to weight loss by up-regulating energy metabolism and the expression of UCP-1 in mitochondria. To investigate the up-stream regulators of the expression of UCP-1, recombinant human CNTF (rhCNTF) (0.1, 0.3, 0.9 mg/kg/day s.c.) administered to KK-Ay mice for 30 days resulting in reductions in body weight and perirenal fat mass. In brown adipose tissues, the gene expressions of nuclear respiratory factor (NRF)-1, mitochondrial transcription factor A (TFam) and uncoupling protein (UCP)-1 were found up regulated by rhCNTF. To the best of our knowledge, these effects represent new insights on the mechanisms of action of weight loss by rhCNTF. In addition, we also found that rhCNTF increased the activity of mitochondrial complex IV. The stimulation of NRF-1, TFam, UCP-1 and the enhanced activity of mitochondrial complex IV may be associated with remedying obesity. The result indicates that rhCNTF can enhance the expressions of NRF-1 and TFam, both of which can up regulate the expression of UCP-1. PMID- 17397830 TI - 1,25-dihydroxyvitamin D3 induces biphasic NF-kappaB responses during HL-60 leukemia cells differentiation through protein induction and PI3K/Akt-dependent phosphorylation/degradation of IkappaB. AB - 1,25-dihydroxyvitamin D(3) (VD(3)) induces differentiation in a number of leukemia cell lines and under various conditions is able to either stimulate or inhibit nuclear factor kappa B (NF-kappaB) activity. Here we report a time dependent biphasic regulation of NF-kappaB in VD(3)-treated HL-60 leukemia cells. After VD(3) treatment there was an early approximately 4 h suppression and a late 8-72 h prolonged reactivation of NF-kappaB. The reactivation of NF-kappaB was concomitant with increased IKK activities, IKK-mediated IkappaBalpha phosphorylation, p65 phosphorylation at residues S276 and S536, p65 nuclear translocation and p65 recruitment to the NF-kappaB/vitamin D responsive element promoters. In parallel with NF-kappaB stimulation, there was an up-regulation of NF-kappaB controlled inflammatory and anti-apoptotic genes such as TNFalpha, IL 1beta and Bcl-xL. VD(3)-triggered reactivation of NF-kappaB was associated with PI3K/Akt phosphorylation. PI3K/Akt antagonists suppressed VD(3)-stimulated IkappaBalpha phosphorylation as well as NF-kappaB-controlled gene expression. The early approximately 4 h VD(3)-mediated NF-kappaB suppression coincided with a prolonged increase of IkappaBalpha protein which require de novo protein synthesis, lasted for as least 72 h and was insensitive to MAPK, IKK or PI3K/Akt inhibitors. Our data suggest a novel biphasic regulation of NF-kappaB in VD(3) treated leukemia cells and our results may have provided the first molecular explanation for the contradictory observations reported on VD(3)-mediated immune regulation. PMID- 17397831 TI - Exposure to a dry environment induces strain-specific responses in mice. AB - Most current animal models of dry eye have a single causative mechanism and do not take into consideration the influence of environmental conditions on tear secretion and associated ocular surface signs. Since immunity and inflammation have been implicated in dry eye pathogenesis, and different mouse strains are known to have differentially biased immune responses, we conducted the present study to test the hypothesis that strains with specifically polarized T cell responses (T helper-1 [Th1] vs. T helper-2 [Th2]) develop differential signs of dry eye when exposed to a controlled low humidity setting. Eight to 12-week-old BALB/c (Th2 biased) and C57BL/6 (Th1 biased) mice were placed in a controlled environment chamber (CEC) where relative humidity (RH), temperature (T), and air flow (AF) were continuously regulated and monitored. Mice were exposed to specific environmental controlled conditions (RH=15.5+/-3.8%, AF=15 l/min, T=21 23 degrees C) for 3 to 7 days. Aqueous tear production by means of the cotton thread test, corneal fluorescein staining (NEI grading scheme, score 0-15) and goblet cell density in the superior and inferior conjunctivae were measured by a masked observer. No statistically significant differences between the groups were found at baseline. Statistically significant decreases in tear secretion were seen after exposure to the CEC environment. Mean cotton thread wetting was 1.9+/ 0.2 (baseline), 1.4+/-0.3 (day 3), and 0.9+/-0.2 mm (day 7) for BALB/c mice, and 1.7+/-0.3 (baseline), 0.9+/-0.3 (day 3), and 0.4+/-0.2 mm (day 7) for C57BL/6 mice. These mice showed reduced tear secretion as compared to BALB/C at each time point tested (P<.005, t-test). Both BALB/c and C57BL/6 mice showed a significant increase in corneal fluorescein staining at both day 3 and day 7 as compared to baseline. With exposure to the CEC goblet cell density significantly decreased in the superior and inferior conjunctivae in BALB/c mice, while it remained unchanged in C57BL/6 mice. This study indicates that exposure of non pharmacologically modified mice to a low humidity environment in the CEC can lead to significant alterations in tear secretion, goblet cell density, and acquisition of dry eye-related ocular surface signs which are strain-specific. PMID- 17397832 TI - Differential dependence of regulatory volume decrease behavior in rabbit corneal epithelial cells on MAPK superfamily activation. AB - We characterized the dependence of hypotonicity-induced regulatory volume decrease (RVD) responses on mitogen-activated protein kinase (MAPK) pathway signaling in SV40-immortalized rabbit corneal epithelial cells (RCEC). Following calcein-AM loading, RVD was monitored using a microplate fluorescence reader. Western blot analysis determined MAPK activation. After 30 min, the RVD response restored the relative cell volume to nearly isotonic values, whereas it was inhibited when cells were bathed either in a Cl- -free solution or with the Cl- channel inhibitors: 5-nitro-2-(3-phenylpropylamino)benzoic acid or niflumic acid. Similar declines occurred with either a high-K+ (20 mM) supplemented solution or the K+ channel inhibitor 4-aminopyridine. Activation of extracellular signal regulated kinase (ERK), p38, and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) was time and tonicity-dependent. Stimulation of ERK and SAPK/JNK was maximized earlier than that of p38. Activation of ERK and SAPK/JNK was insensitive to Cl- and K+ channel inhibitors, whereas inhibition with either PD98059 or SP600125, respectively, blocked RVD. However, inhibition of p38 with SB203580had no effect on RVD. Suppression of RVD instead blocked p38 activation. Differences in the dependence of RVD activation on Erk1/2 and p38 signaling were validated in dominant negative (d/n)-Erk1 and d/n-p38 cells. Volume-sensitive Cl- and K+ channel activation contributes, in concert, to RVD in RCEC. Therefore, swelling-induced ERK and SAPK/JNK stimulation precedes Cl- and K+ channel activation, whereas p38 activation occurs as a consequence of RVD. PMID- 17397833 TI - The sarcoglycan complex in Schwann cells and its role in myelin stability. AB - Sarcoglycans are originally identified in muscle for their involvement in limb girdle muscular dystrophies. They form a multi-meric complex (alpha-, beta-, gamma-, delta-sarcoglycan) that associates with dystrophin, dystroglycan and other proteins to constitute the larger dystrophin-glycoprotein complex at the muscle membrane. Three sarcoglycan subunits (epsilon-, beta-, delta-sarcoglycan) were previously identified in Schwann cells and shown to associate with dystroglycan and a Schwann cell-specific dystrophin isoform (Dp116) at the outermost Schwann cell membrane. Currently, little is known about the exact composition and function of the sarcoglycan complex in the peripheral nervous system. In this study, we showed that the Schwann cell sarcoglycan complex consists of epsilon-, beta-, delta-sarcoglycan and the newly identified zeta sarcoglycan subunit. The expression of sarcoglycans precedes the onset of myelination and is induced by neurons. In sarcoglycan-deficient BIO14.6 hamsters, loss of the Schwann cell sarcoglycan complex reduces the steady state levels of alpha-dystroglycan and Dp116. Ultrastructural analysis of sciatic nerves from the mutant animals revealed altered myelin sheaths and disorganized Schmidt-Lanterman incisures indicative of myelin instability. The disruption in myelin structure increased in severity with age. Nerve conduction studies also showed subtle electrophysiological abnormalities in the BIO14.6 hamsters consistent with reduced myelin stability. Together, these findings suggest an important role of sarcoglycans in the stability of peripheral nerve myelin. PMID- 17397834 TI - Rapid glia expression and release of proinflammatory cytokines in experimental Klebsiella pneumoniae meningoencephalitis. AB - The host immune/inflammatory response following CNS infection by Klebsiella pneumoniae remains poorly understood. Using a rat model of K. pneumoniae meningoencephalitis, we investigated the temporal profiles of brain proinflammatory cytokines and their cellular sources. Leukocyte counts significantly increased in cerebrospinal fluid (CSF) at 2 h after K. pneumoniae inoculation into the rat brain but were still much lower than blood leukocyte counts. However, concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and IL-6 in CSF were much higher than the simultaneously collected serum levels. The rapid increase in brain expression of these cytokines at the messenger RNA (mRNA) and protein levels occurred earlier than the onset of leukocytosis. Double immunofluorescence staining revealed the presence of TNF-alpha, IL-1beta, and IL-6 in astrocytes and microglia. Exposure of primary culture of glial cells to K. pneumoniae also resulted in time dependent increases in the concentration of these cytokines in the culture media. Taken together, our results suggest that glial cells are an important early source of proinflammatory cytokines during K. pneumonia infection of CNS. PMID- 17397835 TI - A novel method of head fixation for the study of rodent facial function. AB - The rodent vibrissial system offers an excellent model for the study of both sensory and motor function. It has been widely employed to gather data pertaining to sensory and motor function involving the 5th and 7th cranial nerves and the central nervous system. Existing methods of head fixation for precise measurements of ocular and vibrissial function involve exposing the cranium and applying dental cement from which two or more threaded rods emerge. This common approach is suboptimal, requiring a relatively complicated implantation procedure, and results in a large, chronic interface between the scalp and environmentally exposed implant material attached to the skull. Here we describe a head fixation device that is inexpensive, easy to build, less prone to infection, preserves access to the cranial midline, and permits repeated measurements over many months. PMID- 17397836 TI - Potassium transporters in plants--involvement in K+ acquisition, redistribution and homeostasis. AB - Potassium is a major plant nutrient which has to be accumulated in great quantity by roots and distributed throughout the plant and within plant cells. Membrane transport of potassium can be mediated by potassium channels and secondary potassium transporters. Plant potassium transporters are present in three families of membrane proteins: the K(+) uptake permeases (KT/HAK/KUP), the K(+) transporter (Trk/HKT) family and the cation proton antiporters (CPA). This review will discuss the contribution of members of each family to potassium acquisition, redistribution and homeostasis. PMID- 17397837 TI - Molecular mechanisms of ammonium transport and accumulation in plants. AB - The integral membrane proteins of the ammonium transporter (AMT/Rh) family provide the major route for shuttling ammonium (NH(4)(+)/NH(3)) across bacterial, archaeal, fungal and plant membranes. These proteins are distantly related to the Rh (rhesus) glycoproteins, which are absent in higher plants, but are present in many species, including bacteria and mammals. It appears that the large nitrogen requirement of plants resulted in unique strategies to acquire, capture and/or release ammonium. The biological function of plant ammonium transporters will be discussed and compared to other AMT/Rh proteins. PMID- 17397839 TI - Increased risk of depressive disorders in women with polycystic ovary syndrome. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) is associated with several metabolic complications. A few small studies have also suggested an increased risk of depression in women with PCOS. The goals of this study were to estimate the prevalence of depressive disorders in women with PCOS compared with controls and to evaluate the correlation between depression, hyperandrogenism, and other metabolic markers. DESIGN: Cohort study. SETTING: University Hospital. PATIENT(S): Women with PCOS (Rotterdam criteria; n = 103). Women without PCOS seen during the same time period for an annual exam were used as control subjects (n = 103). INTERVENTION(S): Primary Care Evaluation of Mental Disorders Patient Health Questionnaire (PRIME-MD PHQ) and the Beck Depression Inventory. MAIN OUTCOME MEASURE(S): Depressive disorders. RESULT(S): Women with PCOS were at an increased risk for depressive disorders (new cases) compared with controls (21% vs. 3%; odds ratio 5.11 [95% confidence interval (CI) 1.26-20.69]; P<.03). The overall risk of depressive disorders in women with PCOS was 4.23 (95% CI 1.49 11.98; P<.01) independent of obesity and infertility. Compared with the nondepressed PCOS subjects, the depressed PCOS subjects had a higher body mass index (BMI) and evidence of insulin resistance (P<.02). CONCLUSION(S): We report a significantly increased risk of depressive disorders (as defined by the Diagnostic and Statistical Manual IV) in women with PCOS and recommend routine screening in this population. PMID- 17397838 TI - Cellular functions of NSF: not just SNAPs and SNAREs. AB - N-ethylmaleimide sensitive factor (NSF) is an ATPases associated with various cellular activities protein (AAA), broadly required for intracellular membrane fusion. NSF functions as a SNAP receptor (SNARE) chaperone which binds, through soluble NSF attachment proteins (SNAPs), to SNARE complexes and utilizes the energy of ATP hydrolysis to disassemble them thus facilitating SNARE recycling. While this is a major function of NSF, it does seem to interact with other proteins, such as the AMPA receptor subunit, GluR2, and beta2-AR and is thought to affect their trafficking patterns. New data suggest that NSF may be regulated by transient post-translational modifications such as phosphorylation and nitrosylation. These new aspects of NSF function as well as its role in SNARE complex dynamics will be discussed. PMID- 17397840 TI - Techniques for removal of the Essure hysteroscopic tubal occlusion device. AB - OBJECTIVE: To present two case reports of techniques for removal of the Essure (Conceptus, Inc., Mountain View, CA) hysteroscopic tubal occlusion device. DESIGN: Case report. SETTING: University-affiliated teaching hospital. PATIENT(S): Two women requesting removal of Essure devices because of persistent pelvic pain up to 6 weeks after placement. INTERVENTION(S): Hysteroscopic and laparoscopic removal of the Essure microinserts. No institutional review board approval was obtained because the procedures were not part of a study protocol. MAIN OUTCOME MEASURE(S): Effective removal of the Essure device and resolution of pelvic pain symptoms. RESULT(S): Successful removal of the device was achieved in both patients. In one case, the procedure was performed entirely by hysteroscopy. In the other case, a laparoscopic approach was required because of a lack of visualization of the device. Pain symptoms resolved within 2 weeks of removal in both patients. CONCLUSION(S): These are the first reported cases of successful removal of the Essure tubal occlusion devices up to 6 weeks after placement. This procedure can be safely performed with the use of hysteroscopy if the inserts are clearly visualized. Laparoscopy is an alternative when the device is completely within the fallopian tube. Further study is needed to assess the functionality of the tube after the procedure, as well as feasibility of removal beyond 6 weeks. PMID- 17397841 TI - Polypectomy devices. PMID- 17397842 TI - Re: avulsion fractures of the lateral tibial condyle in children. PMID- 17397843 TI - The German Multicentre Pelvis Registry: a template for an European Expert Network? AB - The range of severity of pelvic injuries is wide and can include simple, undisplaced pelvic fractures, which may limit the activity of the individual patient for only a short period of time, and severe, complex or even open pelvic fractures, causing immediate life threatening situations. Even with continuous progress in development of techniques and treatment protocols, primary treatment and definitive reconstruction of pelvic ring injuries and acetabular fractures there is still an ongoing debate about specific problems in the evaluation of injuries and fractures. Because of the low incidence of pelvic fractures (37/100,000) the individual experience, which can be acquired by the surgical team, even in major Trauma Centres, is limited and can only be acquired over a longer period of time. The German Multicentre Pelvic Study Group started with reporting of pelvic fractures in 1991 and included 10 University- and Major Trauma Hospitals. The intense work on definitions and classification during the first years generated a universal "language" of understanding, which also helped in unifying indications and even procedures in pelvic and acetabular fractures. With several modifications and expansion of the number of participating hospitals the Group has been active until now and is just entering a "third phase" converting into the "German Multicentre Pelvic and Acetabular Registry" being technologically modified to an Internet based data registry. As this registry is already designed as an open platform, not limited in capacity and regions, it provides a platform, which may easily be expanded to the European level allowing for international multicentre studies and case sampling. Therefore this type of pelvic registry could act as a basis for further scientific evaluation of specific topics in the field of pelvic and acetabular surgery and could be a template for a European Expert Network. Driven by the differences of healthcare systems and organisation of trauma care within Europe and the challenge that pelvic fractures not only can lead to permanent disability, but also play an important role in posttraumatic fatalities, a clear need can be shown for detailed analysis of the present situation within the different European nations. PMID- 17397844 TI - Evaluation and treatment of pelvic metastases. AB - Advances in systemic treatment of cancer have improved patients' survival and increased the number of patients presenting with metastases of the pelvic ring. Pelvic metastatic lesions may cause severe pain and functional disability. A multidisciplinary approach is fundamental for the management of these lesions. Lesions of the pelvis not directly involving the hip joint such as the ischium, pubis or sacroiliac area can generally be treated non-operatively with radiation alone or using minimally invasive procedures of radiofrequency ablation, cryosurgery and percutaneous osteoplasty. Periacetabular destructive lesions may require total hip replacement with reconstruction of the acetabulum dependent on the extent of the defect. Operative treatment should restore the mechanical stability of the hip joint, and preserve the mobility, independence and comfort of these patients. PMID- 17397845 TI - Pelvic and acetabular surgery within Europe: the need for the co-ordination of treatment concepts. AB - Pelvic and acetabular injuries are rare and represent the tip of the trauma iceberg. They often present with other associated injuries. Their management can pose difficulties even to the most experienced trauma surgeons and well-developed trauma systems. Despite the advancements made after the 1960s due to Judet and Letournel's contributions, the pelvic and acetabular sub-specialty has had difficulty in consolidating experience and developing realistic treatment protocols. By means of sharing ideas and results, "learning curves" of individuals and nations could be shortened. As a result, better health quality and advanced medical facilities for our future patients may be anticipated. In this article we examine the current problems affecting the provision of a high quality pelvic and acetabular service and analyse the needs for the co-ordination of treatment concepts within the European Landscape. PMID- 17397846 TI - Is there a role for percutaneous pelvic and acetabular reconstruction? AB - The primary goal in the treatment of pelvic fractures is the restoration of haemodynamic stability. The secondary goal is the reconstruction of stability and symmetry of the pelvic ring. Percutaneous reconstruction can only be accepted if these goals are met. The type of definitive surgery is dependent of the degree of instability of the anterior and posterior pelvic ring. Retrograde transpubic screw fixation of pubic rami fractures is a good alternative to external fixation or plate and screw osteosynthesis. The technique of screw placement and image intensifier control is explained. Internal fixation of pure sacroiliac dislocations, fracture-dislocations of the sacroiliac joint and sacral fractures can be fixed with sacroiliac screws, placed percutaneously. Reduction of the fracture or dislocation is performed closed, or open if anatomy cannot be restored in a closed manner. The primary goal in the treatment of acetabular fractures is to restore anatomy. Reduction comes before fixation. The goal of minimising approaches cannot be more important. In most cases open reduction will be necessary to achieve anatomical reconstruction. Only the experienced acetabular surgeon will be able to decide when and how he can restore anatomy through a less invasive approach or with a percutaneous procedure. The anterior column screw can be inserted through a separate incision in addition to a Kocher Langenbeck approach. It is the same screw as the retrograde transpubic screw but placed in the opposite direction. The posterior column screw is placed percutaneously from the lateral cortex of the ilium in the direction of the posterior column. Techniques of placement of both screws are demonstrated. Open reduction and internal fixation remains the standard of care in stabilisation of pelvic and acetabular fractures. Only the experienced surgeon will be able to judge if percutaneous procedures can be an alternative or a useful additive to conventional techniques. PMID- 17397847 TI - Candida albicans growth inhibition by tumor necrosis factor-alpha in vaginal epithelial cells. PMID- 17397848 TI - Sigmoid volvulus in the puerperium. PMID- 17397849 TI - Tasks performance by registered nurses and care assistants in nursing homes: a quantitative comparison of survey data. AB - OBJECTIVES: To examine the care tasks performed for residents of Belgian care institutions for older people and the association between the performance of care tasks by persons of different staffing categories (registered nurses (RN) and care assistants (CA)) and the characteristics (the dependency level and the diagnosis of dementia) of the residents. STUDY DESIGN AND SETTING: Survey on the care for residents, aged 65 or over, living in 26 care institutions for aged people. MAIN OUTCOME MEASURES: Separate bivariate comparisons of care time spent on residents with and without dementia and bivariate comparisons of care time spent on six task categories on residents of different dependency levels were performed. The median number of minutes over seven days (mosd) and the inter quartile range (IQR) are presented as summary measures. The proportion of the time spent by RNs and CAs per resident and per task category was calculated. RESULTS: Time spent was highest on primary care tasks (34,554 mosd; 48.7%), followed by in order of time spent, supportive tasks (10,845 mosd; 15.3%), logistic tasks (10,697 mosd; 15.1%), practical nursing procedures (8,689 mosd; 12.2%), administrative tasks (3,357 mosd; 4.7%) and communication tasks (2,814 mosd; 4.0%). Overall there was no significant difference between the total time spent by RNs (median=190 mosd; IQR 105-334) and the total time spent by CAs (median=196 mosd; IQR 91-331; p=0.89). RNs were spending significantly more time than CAs in practical nursing procedures, communication tasks and administrative tasks. CONCLUSIONS: There was a sharp task demarcation between RNs and CAs in the three less frequent task categories. There was no indication that RNs were delegating tasks to CAs. PMID- 17397850 TI - Reduction of atherosclerotic nanoplaque formation and size by Ginkgo biloba (EGb 761) in cardiovascular high-risk patients. AB - Coating a silica surface with the isolated lipoprotein receptor proteoheparan sulfate (HS-PG) from arterial endothelium and vascular matrices and adding both the atherogenic VLDL/IDL/LDL lipid fraction in its native composition and Ca(2+) ions, we could observe in vitro the earliest stages of atherosclerotic plaque development by ellipsometric techniques (patent EP 0 946 876). This so-called nanoplaque formation is represented by the ternary aggregational complex of the HS-PG receptor, lipoprotein particles and calcium ions. The model was validated in several clinical studies on statins in cardiovascular high-risk patients. In eight patients who had undergone an aortocoronary bypass operation, the reduction of atherosclerotic nanoplaque formation amounted to 11.9+/-2.5% (p<0.0078) and of nanoplaque size to 24.4+/-8.1% (p<0.0234), respectively, after a 2-month therapy with Ginkgo biloba extract (2x 120 mg daily, EGb 761). Additionally, superoxide dismutase (SOD) activity was upregulated by 15.7+/-7.0% (p<0.0391), the quotient oxLDL/LDL lowered by 17.0+/-5.5% (p<0.0234) and lipoprotein(a) concentration decreased by 23.4+/-7.9% (p<0.0234) in the patients' blood. The concentration of the vasodilating substances cAMP and cGMP was augmented by 37.5+/-9.1% (p<0.0078) and 27.7+/-8.3% (p<0.0156), respectively. A multiple regression analysis between the patients' VLDL/IDL/LDL lipoprotein fraction applied in the ellipsometry measurements as well as the further risk factors oxLDL/LDL and Lp(a) on the one hand and changes in nanoplaque formation on the other hand reveals a basis for a mechanistic explanation of nanoplaque reduction under ginkgo treatment. The atherosclerosis inhibiting effect is possibly due to an upregulation in the body's own radical scavenging enzymes and an attenuation of the risk factors oxLDL/LDL and Lp(a). PMID- 17397851 TI - Zonal changes in the three-dimensional morphology of the chondron under compression: the relationship among cellular, pericellular, and extracellular deformation in articular cartilage. AB - The pericellular matrix (PCM) is a narrow region of tissue that completely surrounds chondrocytes in articular cartilage. Previous theoretical models of the "chondron" (the PCM with enclosed cells) suggest that the structure and properties of the PCM may significantly influence the mechanical environment of the chondrocyte. The objective of this study was to quantify changes in the three dimensional (3D) morphology of the chondron in situ at different magnitudes of compression applied to the cartilage extracellular matrix. Fluorescence immunolabeling for type-VI collagen was used to identify the boundaries of the cell and PCM, and confocal microscopy was used to form 3D images of chondrons from superficial, middle, and deep zone cartilage in explants compressed to 0%, 10%, 30%, and 50% surface-to-surface strain. Lagrangian tissue strain, determined locally using texture correlation, was highly inhomogeneous and revealed depth dependent compressive stiffness and Poisson's ratio of the extracellular matrix. Compression significantly decreased cell and chondron height and volume, depending on the zone and magnitude of compression. In the superficial zone, cellular-level strains were always lower than tissue-level strains. In the middle and deep zones, however, tissue strains below 25% were amplified at the cellular level, while tissue strains above 25% were decreased at the cellular level. These findings are consistent with previous theoretical models of the chondron, suggesting that the PCM can serve as either a protective layer for the chondrocyte or a transducer that amplifies strain, such that cellular-level strains are more homogenous throughout the tissue depth despite large inhomogeneities in local ECM strains. PMID- 17397852 TI - Application of fractional factorial experimental and Box-Behnken designs for optimization of single-drop microextraction of 2,4,6-trichloroanisole and 2,4,6 tribromoanisole from wine samples. AB - In this paper a new method for the determination of 2,4,6-trichloroanisole (TCA) and 2,4,6-tribromoanisole (TBA) in wine samples is presented. Headspace single drop microextraction (HS-SDME) was used for the extraction and preconcentration of the analytes, followed by analysis by gas chromatography and electron-capture detection (GC-ECD). The variables affecting extraction efficiency were optimized using fractional factorial experimental and Box-Behnken designs. The external calibration procedure was successfully carried out using a synthetic wine solution and diluted red wine samples. The method was also applied to white wine samples. Excellent detection limits of 8.1 and 6.1 ng L(-1) were achieved for TCA and TBA, respectively. Good precision and accuracy were obtained. PMID- 17397853 TI - Hydrophobic and electrostatic forces control the retention of membrane peptides and proteins with an immobilised phosphatidic acid column. AB - The retention behaviour of four membrane-associated peptides and proteins with an immobilized phosphatidic acid (PA) stationary phase was evaluated. The solutes included the cytolytic peptides gramicidin A and melittin, the integral membrane protein bacteriorhodpsin and cytochrome c, a peripheral membrane protein. Gramicidin has no nett charge and exhibited normal reversed phase-like behaviour which was largely independent of mobile phase pH. In contrast, melittin, which has a positively charged C-terminal tail, exhibited reversed phase like retention at pH 5.4 and 7.4, and was not retained at pH 3 reflecting the influence of electrostatic interactions with the negatively charged phosphatidic acid ligand. Bacteriorhodpsin was eluted at high acetonitrile concentrations at pH 3 and 5.4 and cytochrome c was only eluted at pH 3. Moreover, cytochrome c eluted in the breakthrough peak between 0 and 100% acetonitrile, demonstrating the role of electrostatic interactions with the PA surface. Overall, the results demonstrate that pH can be used to optimize the fractionation and separation of membrane proteins with immobilized lipid stationary phases. PMID- 17397855 TI - Systematic errors in analytical measurement results. AB - Definitions of the concepts of bias and recovery are discussed and approaches to dealing with them described. The Guide To Uncertainty in Measurement (GUM) recommends correction for all significant systematic effects, but it is also possible to expand measurement uncertainty to take account of uncorrected bias. Run, laboratory and method bias can be defined as components of the bias of a particular measurement result, and can be useful as concepts used in method validation. Estimation of run bias allows a simplification of the estimation of measurement uncertainty. Multivariate calibration brings its own biases that must be quantified and minimised. PMID- 17397856 TI - Effects of dissolved polymer on the transport of colloidal particles through a microcapillary. AB - The effect of water-soluble polymer on the transport of latex particles through a microcapillary was investigated. Capillary hydrodynamic fractionation (CHDF) experiments were performed using polystyrene (PS) particles and poly(ethylene oxide) (PEO) solutions as the eluant. Generally, the average particle velocities were greater than those corresponding to a polymer-free eluant. A decrease in the sample axial dispersion was also observed using the PEO solutions. In addition, increasing the polymer molecular weight resulted in lower particle residence times in the capillary tube. The enhanced particle transport arises primarily from an increase in the particle diameter resulting from the adsorption of PEO onto the PS surfaces, and, more importantly, from the migration of particles toward the capillary axis due to the normal stress of the PEO solution. PMID- 17397857 TI - Preparation and characterization of novel cationic pH-responsive poly(N,N' dimethylamino ethyl methacrylate) microgels. AB - Novel monodisperse cationic pH-responsive microgels were successfully prepared by dispersion polymerization in ethanol/water mixture using N,N'-dimethylamino ethyl methacrylate (DMAEMA) as the monomer, poly(vinyl pyrrolidone) (PVP) as the steric stabilizer and N,N'-methylenebisacrylamide (MBA) as the cross-linker. The effects of various polymerization parameters, such as medium polarity, concentration of cross-linker, concentration of monomer, and concentration and molecular weight of stabilizer on the final diameter and monodispersity of poly(N,N'-dimethylamino ethyl methacrylate) (PDMAEMA) microgels were systematically studied. The pH responsive characteristics of PDMAEMA microgels were also investigated. The experimental results showed that these microgels exhibited excellent pH responsivity and significantly swelled at low pH values. The maximum ratio of volume change of the prepared microgels in response to pH variation was more than 11 times. It was found that the prepared microgels completely aggregated at the isoelectric point (IEP) around pH 6. On the other hand, the microgels were stable in aqueous solution at both low and high pH values. The results can be used for effectively controlled separation of particles. PMID- 17397858 TI - Characterization of cytoskeletal proteins in follicular structures of cows with cystic ovarian disease. AB - The distribution of intermediate filaments (vimentin, cytokeratins, desmin) and microfilaments (alpha-smooth muscle actin and muscle specific actin) was studied immunohistochemically in bovine ovaries, with and without cystic ovarian disease. The immunohistochemically stained area (IHCSA), was quantified by image analysis, to evaluate the expression of these cytoskeletal proteins in the follicular wall of healthy antral, atretic, and cystic follicles. The granulosa cell layer of cystic follicles and atretic follicles had a significantly larger IHCSA for vimentin than did healthy antral follicles. Cytokeratins reacted lightly in the granulosa cells of antral follicles of normal ovaries, whereas granulosa cells of atretic and cystic follicles showed significantly higher IHCSA values. Immunohistochemical localization of desmin, muscle specific actin, and alpha smooth muscle actin was restricted to the theca externa. This study supports earlier suggestions that strongly positive reactions with vimentin and cytokeratin antibodies observed in the granulosa cells of cystic follicles are due to the reorganization that occurs in the follicle during the process of cystic development, and are associated with changes in the expression of cytoskeletal proteins that are essential to proper cellular functioning. PMID- 17397859 TI - Comparison of promoter activities for efficient expression into human B cells and haematopoietic progenitors with adenovirus Ad5/F35. AB - Adenoviral gene transfer into human B lymphocytes and haematopoietic progenitors would allow the characterization of their function on cellular growth, differentiation and survival. Efficient gene expression is however strongly dependent on the promoter used. In this study, we investigated the relative strength of various promoters by following and measuring the expression of the reporter gene EYFP in human peripheral B lymphocytes, cord blood CD34(+) cells and the megakaryocytic cell line M-07e. The murine PGK promoter provided the best level of transgene expression in CD34(+) cells among the four promoters tested, followed closely by the CMV promoter, and to a lesser extend by a CMV promoter with a beta-globin/IgG chimeric intron, whereas the human CD40 promoter provided the lowest levels of expression. In contrast, the strongest promoters in B lymphocytes were the two CMV promoters. Surprisingly, even the best promoters were unable to induce transgene expression in more than 75-80% of the primary B and CD34(+) cells, even though 100% of the cells were infected. Finally and in contrast to retroviruses, only a minority of B lymphocytes and CD34(+) cells were able to induce the transcription of IRES-containing bicistronic expression cassettes from adenovirus. PMID- 17397860 TI - Ischemic preconditioning involves dual cardio-protective axes with p38MAPK as upstream target. AB - The existing literature indicates a crucial role of p38 MAP (mitogen-activated protein) kinase (p38MAPK) and its downstream target MAPKAP kinase 2 (MK2) in ischemic preconditioning (IPC). Accordingly, deletion of MK2 gene should abolish the cardioprotective ability of IPC. Interestingly, we were able to partially precondition the hearts from MK2(-/-) knockout mice suggesting the existence of an as yet unknown alternative downstream target of p38MAPK. A recent study from our laboratory also determined a crucial role of CREB (cyclic AMP response element binding protein) in IPC. Since CREB is a downstream target of MSK-1 (mitogen- and stress-activated protein kinase-1) situated at the crossroad of ERK (extracellular receptor kinase) and p38MAPK signaling pathways, we reasoned that MSK-1 could be a downstream molecular target for p38MAPK and ERK signaling in the IPC hearts. To test this hypothesis, the rat hearts were subjected to IPC by four cyclic episodes of 5 min ischemia and 10 min reperfusion. As expected, IPC induced the activation of ERK1/2, p38MAPK, MK2 and HSP (heat shock protein) 27 as evidenced by their increased phosphorylation; and the inhibition of p38MAPK with SB203580 almost completely, and the inhibition of ERK1/2 with PD098059 partially, abolished cardioprotective effects of IPC. Inhibition of MSK-1 with short hairpin RNA (shRNA) also abolished the IPC-induced cardioprotection. SB203580 partially blocked the effects of MSK-1 suggesting that MSK-1 sits downstream of p38MAPK. shRNA-MSK-1 blocked the contribution of both p38MAPK and ERK1/2 as it is uniquely situated at the downstream crossroad of both of these MAP kinases. Although MSK-1 sits downstream of both ERK1/2 and p38MAPK, ERK1/2 activation appears to play less significant role compared to p38MAPK, since its inhibition blocked MSK activation only partially. Consistent with these results, shRNA-MSK-1 blocked the partial PC in MK2(-/-) hearts, and in combination with SB203580, completely abolished the PC effects in the wild-type hearts. The IPC-induced survival signaling was almost completely inhibited with SB203580, and only partially with PD 098059 as evidenced from the inhibition patterns of IPC induced activation of CREB, Akt and Bcl-2. Again SB203580 alone or in combination with shRNA-MSK-1 inhibited IPC induced survival signal comparatively, suggesting that MSK-1 exists downstream of p38MAPK. Taken together, these results indicate for the first time MSK-1 as an alternative (other than MK2) downstream target for p38MAPK, which also transmits survival signal through the activation of CREB. PMID- 17397861 TI - The structure of a tandem pair of spectrin repeats of plectin reveals a modular organization of the plakin domain. AB - Plectin is a large and versatile cytoskeletal linker and member of the plakin protein family. Plakins share a conserved region called the plakin domain located near their N terminus. We have determined the crystal structure of an N-terminal fragment of the plakin domain of plectin to 2.05 A resolution. This region is adjacent to the actin-binding domain and is required for efficient binding to the integrin alpha6beta4 in hemidesmosomes. The structure is formed by two spectrin repeats connected by an alpha-helix that spans these two repeats. While the first repeat is very similar to other known structures, the second repeat is structurally different with a hydrophobic core, narrower than that in canonical spectrin repeats. Sequence analysis of the plakin domain revealed the presence of up to nine consecutive spectrin repeats organized in an array of tandem modules, and a Src-homology 3 domain inserted in the central spectrin repeat. The structure of the plakin domain is reminiscent of the modular organization of members of the spectrin family. The architecture of the plakin domain suggests that it forms an elongated and flexible structure, and provides a novel molecular explanation for the contribution of plectin and other plakins to the elasticity and stability of tissues subjected to mechanical stress, such as the skin and striated muscle. PMID- 17397862 TI - The Alzheimer's peptides Abeta40 and 42 adopt distinct conformations in water: a combined MD / NMR study. AB - The role of peptides Abeta40 and Abeta42 in the early pathogenesis of Alzheimer's disease (AD) is frequently emphasized in the literature. It is known that Abeta42 is more prone to aggregation than Abeta40, even though they differ in only two (IA) amino acid residues at the C-terminal end. A direct comparison of the ensembles of conformations adopted by the monomers in solution has been limited by the inherent flexibility of the unfolded peptides. Here, we characterize the conformations of Abeta40 and Abeta42 in water by using a combination of molecular dynamics (MD) and measured scalar (3)J(HNHalpha) data from NMR experiments. We perform replica exchange MD (REMD) simulations and find that classical forcefields reproduce the NMR data quantitatively when the sampling is extended to the microseconds time-scale. Using the quantitative agreement of the NMR data as a validation of the model, we proceed to compare the conformational ensembles of the Abeta40 and Abeta42 peptide monomers. Our analysis confirms the existence of structured regions within the otherwise flexible Abeta peptides. We find that the C terminus of Abeta42 is more structured than that of Abeta40. The formation of a beta-hairpin in the sequence (31)IIGLMVGGVVIA involving short strands at residues 31-34 and 38-41 (in bold) reduces the C-terminal flexibility of the Abeta42 peptide and may be responsible for the higher propensity of this peptide to form amyloids. PMID- 17397863 TI - Crystal structure, biochemical and genetic characterization of yeast and E. cuniculi TAF(II)5 N-terminal domain: implications for TFIID assembly. AB - General transcription factor TFIID plays an essential role in transcription initiation by RNA polymerase II at numerous promoters. However, understanding of the assembly and a full structural characterization of this large 15 subunit complex is lacking. TFIID subunit TAF(II)5 has been shown to be present twice in this complex and to be critical for the function and assembly of TFIID. Especially, the TAF(II)5 N-terminal domain is required for its incorporation within TFIID and immuno-labelling experiments carried out by electron microscopy at low resolution have suggested that this domain might homodimerize, possibly explaining the three-lobed architecture of TFIID. However, the resolution at which the electron microscopy (EM) analyses were conducted is not sufficient to determine whether homodimerization occurs or whether a more intricate assembly implying other subunits is required. Here we report the X-ray structures of the fully evolutionary conserved C-terminal sub-domain of the TAF(II)5 N terminus, from yeast and the mammalian parasite Encephalitozoon cuniculi. This sub-domain displays a novel fold with specific surfaces having conserved physico-chemical properties that can form protein-protein interactions. Although a crystallographic dimer implying one of these surfaces is present in one of the crystal forms, several biochemical analyses show that this sub-domain is monomeric in solution, even at various salt conditions and in presence of different divalent cations. Consequently, the N-terminal sub-domain of the TAF(II)5 N terminus, which is homologous to a dimerization motif but has not been fully conserved during evolution, was studied by analytical ultracentrifugation and yeast genetics. Our results show that this sub-domain dimerizes at very high concentration but is neither required for yeast viability, nor for incorporation of two TAF(II)5 molecules within TFIID and for the assembly of this complex. Altogether, although our results do not argue in favour of a homodimerization of the TAF(II)5 N-terminal domain, our structural analyses suggest a role for this domain in assembly of TFIID and its related complexes SAGA, STAGA, TFTC and PCAF. PMID- 17397864 TI - The structural basis for exopolygalacturonase activity in a family 28 glycoside hydrolase. AB - Family 28 glycoside hydrolases (polygalacturonases) are found in organisms across the plant, fungal and bacterial kingdoms, where they are central to diverse biological functions such as fruit ripening, biomass recycling and plant pathogenesis. The structures of several polygalacturonases have been reported; however, all of these enzymes utilize an endo-mode of digestion, which generates a spectrum of oligosaccharide products with varying degrees of polymerization. The structure of a complementary exo-acting polygalacturonase and an accompanying explanation of the molecular determinants for its specialized activity have been noticeably lacking. We present the structure of an exopolygalacturonase from Yersinia enterocolitica, YeGH28 in a native form (solved to 2.19 A resolution) and a digalacturonic acid product complex (solved to 2.10 A resolution). The activity of YeGH28 is due to inserted stretches of amino acid residues that transform the active site from the open-ended channel observed in the endopolygalacturonases to a closed pocket that restricts the enzyme to the exclusive attack of the non-reducing end of oligogalacturonide substrates. In addition, YeGH28 possesses a fused FN3 domain with unknown function, the first such structure described in pectin active enzymes. PMID- 17397865 TI - The crystal structure of a virus-like particle from the hyperthermophilic archaeon Pyrococcus furiosus provides insight into the evolution of viruses. AB - Pyrococcus furiosus is a hyperthermophilic archaeal microorganism found near deep sea thermal vents and its optimal growth temperature of 100 degrees C. Recently, a 38.8-kDa protein from P. furiosus DSM 3638 was isolated and characterized. Electron microscopy revealed that this protein aggregated as spheres of approximately 30 nm in diameter, which we designated P. furiosus virus-like particles (PfVs). X-ray crystallographic analysis at 3.6-A resolution revealed that each PfV consisted of 180 copies of the 38.8-kDa protein and retained T=3 icosahedral symmetry, as is often the case in spherical viruses. The total molecular mass of each particle was approximately 7 MDa. An examination of capsid structures suggested strong evolutionary links among PfV, tailed double-stranded DNA bacteriophages, and herpes viruses. The similar three-dimensional structures of the various coat proteins indicate that these viral capsids might have originated and evolved from a common ancestor. The structure of PfV provides a previously undescribed example of viral relationships across the three domains of life (Eukarya, Bacteria, and Archaea). PMID- 17397866 TI - Comparison of proteolytic susceptibility in phosphoglycerate kinases from yeast and E. coli: modulation of conformational ensembles without altering structure or stability. AB - Escherichia coli phosphoglycerate kinase (PGK) is resistant to proteolytic cleavage while the yeast homolog from Saccharomyces cerevisiae is not. We have explored the biophysical basis of this surprising difference. The sequences of these homologs are 39% identical and 56% similar. Determination of the crystal structure for the E. coli protein and comparison to the previously solved yeast structure reveals that the two proteins have extremely similar tertiary structures, and their global stabilities determined by equilibrium denaturation are also very similar. The extrapolated unfolding rate of E. coli PGK is, however, 10(5) slower than that of the yeast homolog. This surprisingly large difference in unfolding rates appears to arise from a divergence in the extent of cooperativity between the two structural domains (the N and C-domains) that make up these kinases. This is supported by: (1) the C-domain of E. coli PGK cannot be expressed or fold independently of the N-domain, while both domains of the yeast protein fold in isolation into stable structures and (2) the energetics and kinetics of the proteolytically sensitive state of E. coli PGK match those for global unfolding. This suggests that proteolysis occurs from the globally unfolded state of E. coli PGK, while the characteristics defining the yeast homolog suggest that proteolysis occurs upon unfolding of only the C-domain, with the N-domain remaining folded and consequently resistant to cleavage. PMID- 17397867 TI - Insertion of a chaperone domain converts FKBP12 into a powerful catalyst of protein folding. AB - The catalytic activity of human FKBP12 as a prolyl isomerase is high towards short peptides, but very low in proline-limited protein folding reactions. In contrast, the SlyD proteins, which are members of the FKBP family, are highly active as folding enzymes. They contain an extra "insert-in-flap" or IF domain near the prolyl isomerase active site. The excision of this domain did not affect the prolyl isomerase activity of SlyD from Escherichia coli towards short peptide substrates but abolished its catalytic activity in proline-limited protein folding reactions. The reciprocal insertion of the IF domain of SlyD into human FKBP12 increased its folding activity 200-fold and generated a folding catalyst that is more active than SlyD itself. The IF domain binds to refolding protein chains and thus functions as a chaperone module. A prolyl isomerase catalytic site and a separate chaperone site with an adapted affinity for refolding protein chains are the key elements for a productive coupling between the catalysis of prolyl isomerization and conformational folding in the enzymatic mechanisms of SlyD and other prolyl isomerases, such as trigger factor and FkpA. PMID- 17397868 TI - Increased leukotriene c4 synthesis accompanied enhanced leukotriene c4 synthase expression and activities of ischemia-reperfusion-injured liver in rats. AB - BACKGROUND: Hepatic ischemia-reperfusion (I/R) injury is an important clinical issue and relates to cysteinyl leukotrienes (LTs), the first committed synthesis step of which is that LTC4 synthesis enzymes including leukotriene C4 synthase (LTC4S), microsomal glutathione-S-transferase (mGST)2, and mGST3-catalyzed LTA4 and reduced glutathione (GSH), to generate LTC4. However, the mechanisms of LTC4 generation during hepatic I/R are far from being elucidated. MATERIALS AND METHODS: Adult male Sprague Dawley rats were divided into two groups: sham group (control) and I/R group. Liver was subjected to 60 min of partial hepatic ischemia followed by 5 h of reperfusion; saline was administered intravenously. LTC4 content, the activities, and expressions of LTC4 synthesis enzymes were examined with reversed phase high-performance liquid chromatography, reverse transcriptase-polymerase chain reaction, immunoblot, and immunohistochemistry, respectively. Liver damage was assessed by serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) measurements and histological observation. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) level in liver tissue were used to evaluate lipid peroxidation, and oxidative stress was estimated by the reduced GSH level in liver tissue in the pathological process. RESULTS: Compared with control, LTC4 content, the LTC4 synthesis enzymes' activities, and the mRNA and protein expressions of LTC4S were significantly increased, while the mRNA expressions of mGST2 and mGST3 were declined obviously in rat liver during I/R (P < 0.05); most hepatocytes and sinusoidal endothelial cells expressed intensively LTC4S in an I/R-sensitive manner. This was accompanied by the increase in serum ALT and AST levels together with liver tissue MDA content (P < 0.05), the decrease in liver tissue GSH level, and SOD activity (P < 0.05), as well as histological damage. There were no differences in the protein expression of mGST3 between control and I/R groups. CONCLUSIONS: These results demonstrated that hepatic I/R injury up-regulated the mRNA and protein expressions of LTC4S in hepatocytes and sinusoidal endothelial cells and enhanced the activities of the LTC4 synthesis enzymes. It suggests that LTC4 accumulation after hepatic I/R can be caused partially by LTC4S expression up regulation and the LTC4 synthesis enzymes' activities augment to which LTC4S rather than mGST2 or mGST3 may mainly contribute. PMID- 17397869 TI - The effects of iloprost and vitamin C on kidney as a remote organ after ischemia/reperfusion of lower extremities. AB - BACKGROUND: Abdominal aortic surgery can cause ischemic/reperfusion (I/R) injury in not only the lower extremities, but also in the remote organs and tissues such as lungs, kidneys, heart, and liver during abdominal aortic surgery. It can result in mortality and morbidity because of the remote organ injury in early postoperative period. In this study, we investigate the effects of iloprost and vitamin C on the kidney remote organ damage after I/R following abdominal aortic surgery. MATERIAL AND METHODS: Thirty-four adult male Wistar rats were used and divided into five groups. I/R was studied infrarenally in the abdominal aorta following a median laparotomy. The left kidney was excised immediately following the laparotomy in group I (n = 6, normal group). Group II (n = 6) was the sham group. Group III (n = 6, control group) was subjected to 3 h of ischemia followed by an hour of reperfusion. Group IV (n = 8) was given iloprost 20 ng/kg/min during I/R period before aortic-clamping. Group V (n = 8) was given vitamin C 100 mg/kg during I/R period before aortic-clamping. Arterial blood samples were obtained to determine the levels of blood pH, pO(2) (mmHg), pCO2 (mmHg), HCO(3) (mmol/L), and plasma malondialdehyde (MDA, nmol/mL) at the end of reperfusion period in all groups. The left kidneys were used for remote measurements of tissue MDA (nmol/g.w.t) and scored by histopathological examination for acute inflammation. RESULTS: While the arterial blood pO(2) and HCO(3) levels significantly increased, the plasma and renal parenchymal MDA levels significantly decreased in both group IV and group V when compared to group III (P < 0.05). Histopathological and acute inflammation scores statistically decreased in both group IV and V compared with group III (P < 0.05). Although MDA levels, histopathologic and acute inflammation scores in group V were lower than group IV, the differences were not statistically significant (P > 0.05). CONCLUSION: Both iloprost and vitamin C decreased remote organ damage on the kidney after I/R of lower extremities in the rat model. However, vitamin C is more effective than iloprost in preventing postoperative renal dysfunction. PMID- 17397870 TI - Home-anticoagulation testing: accuracy of patient-reported values. AB - BACKGROUND: Few studies have examined the accuracy of patient-reported international normalized (INR) values for home anticoagulation testing (HAT). Our study objectives were to assess this accuracy and compare the percentage time within therapeutic range (PTWTR) based on HAT data to that obtained with testing through an anticoagulation clinic service (ACS). MATERIALS AND METHODS: Forty nine anticoagulated patients were enrolled in a year-long, prospective, crossover study comparing HAT to ACS testing. Patients performed HAT for 6 months and telephoned their INR values. Thereafter, devices were interrogated for the 30 most recent INR readings. Data accuracy was calculated for each patient as the percentage of correctly telephoned INR values divided by the total number of INR values common to both the device and the telephone logs. The device-based PTWTR was compared to the PTWTR based on ACS data. RESULTS: Of the 49 enrolled patients, 32 completed the study protocol. The mean accuracy of reporting was 94.0 +/- 13.0% (range, 48-100%); the median accuracy was 100%. Three patients had marked low accuracy (48, 60, 62%). No significant difference was seen between the PTWTR based on device data versus that obtained though the ACS (59.8 +/- 15.7% versus 59.5 +/- 19.4%, P=0.48). CONCLUSIONS: The overall accuracy of patient reported INR values is high and the PTWTR found with HAT is comparable to that obtained with clinic testing. However, the potential for noncompliance in a small number of patients raises the question of whether periodic confirmation of patient-reported INR values should be considered. PMID- 17397871 TI - Matrix metalloproteinases and inhibitor in knee synovial fluid as cartilage biomarkers in rabbits: the effect of high-intensity jumping exercise. AB - OBJECTIVE: To study the cartilage injury of knee joint in a rabbit model under high-intensity jumping training and to investigate if levels of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-3 (MMP-3), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) in synovial fluid (SF) can be used to predict early sports injury of articular cartilage effectively. MATERIALS AND METHODS: Forty New Zealand white rabbits were divided into two groups randomly: untreated control group (CTRL, n = 8) and jumping training group (TG, n = 32). Concentrations of MMP-1, MMP-3, and TIMP-1 in SF were measured by ELISA assays at 4 and 8 weeks, respectively. Rabbits were euthanized at 4 and 8 weeks, and knee joints were taken out to be examined histologically. Sulfated glycosaminoglycan (GAG) content, thickness of cartilage and subchondral bone, dead cell ratio, and Mankin grades were measured. RESULTS: At 4 weeks, sulfated GAG content, thickness of subchondral bone, and Mankin grades in TG were significantly higher than control. After 8 weeks, the TG cohort had a further increase in the articular cartilage injury. SF levels of MMP-3, TIMP-1, and MMP 3/TIMP-1 in TG were significantly higher than control, and the level of these biomarkers was significantly associated with the severity of the articular cartilage pathology. CONCLUSIONS: Repetitive and high-intensity jumping movement may induce sports injury in the knee joint cartilage. MMP-1, MMP-3, TIMP-1, and MMP-3/TIMP-1 in the SF may accurately predict the severity and pathological characteristic of the joint cartilage injury. PMID- 17397873 TI - Inflammatory demyelination and neurodegeneration in early multiple sclerosis. AB - A number of recent magnetic resonance imaging studies have challenged the classical view of multiple sclerosis (MS) as a "two-stage" disease where an early inflammatory demyelinating phase with focal macroscopic lesions formed in the white matter (WM) of the central nervous system is followed by a late neurodegenerative phase, which is believed to be a mere consequence of repeated inflammatory insults and irreversible demyelination. These studies have consistently shown the presence of diffuse normal-appearing WM damage, marked gray matter involvement and significant cortical functional reorganization, as well as the occurrence of the neurodegenerative component of MS from the earliest clinical stages of the disease with only a partial relation to MRI markers of inflammatory demyelination. The present review argues that MS can no longer be viewed as a "two-stage" disease, which suggests that the two pathological components are dissociated in time, but rather as a "simultaneous two-component" disease, where the relative contributions of the various pathological processes of the disease to the development of "fixed" disability, their relationship and their evolution over time need to be clarified. This new view of MS should inform the development of future research protocols to define its actual physiopathology and prompt the institution of early treatment which should ideally target not only inflammatory demyelination, but also the neurodegenerative aspects of the disease, as well as promote neuroprotection and enhance reparative mechanisms and adaptive functional reorganization of the cortex. PMID- 17397874 TI - Perfect reciprocity is the only evolutionarily stable strategy in the continuous iterated prisoner's dilemma. AB - Theoretical studies have shown that cooperation tends to evolve when interacting individuals have positively correlated phenotypes. In the present article, we explore the situation where this correlation results from information exchange between social partners, and behavioral flexibility. We consider the game 'continuous iterated prisoner's dilemma'. The level of cooperation expressed by individuals in this game, together with their ability to respond to one another, both evolve as two aspects of their behavioral strategy. The conditions for a strategy to be evolutionarily stable in this game are degenerate, and earlier works were thus unable to find a single ESS. However, a detailed invasion analysis, together with the study of evolution in finite populations, reveals that natural selection favors strategies whereby individuals respond to their opponent's actions in a perfectly mirrored (i.e., correlated) fashion. As a corollary, the overall payoff of social interactions (i.e., the amount of cooperation) is maximized because couples of correlated partners effectively become the units of selection. PMID- 17397875 TI - Cysteinyl-leukotriene 1 receptor antagonist attenuates bleomycin-induced pulmonary fibrosis in mice. AB - Leukotrienes are lipid mediators of inflammation derived from the 5-lipoxygenase pathway of arachidonic acid metabolism, and recent evidence suggests that they play an important role in pulmonary fibrosis. Montelukast is a cysteinyl leukotriene 1 receptor antagonist that has been found to reduce airway remodeling, including subepithelial fibrosis, in a murine model of asthma, but the therapeutic effect of montelukast on pulmonary fibrosis remains unclear. In this study, we investigated whether montelukast is capable of preventing bleomycin-induced pulmonary fibrosis in mice. On day 1, C57BL/6 mice were given a single intratracheal injection of bleomycin (2.5 mg/kg), and montelukast (1.0 mg/kg) or vehicle alone subcutaneously 2 h later and on days 1-5 of each week for two weeks. The total number of cells in bronchoalveolar lavage fluid (BALF) was reduced in the montelukast group on day 7 and on day 14, and cellular inflammation and fibrosis were attenuated on day 14 as indicated by significant decrease in the Ashcroft score and lung hydroxyproline content. Although cysteinyl-leukotriene level in BALF was not significantly different, transforming growth factor beta (TGFbeta) level in BALF by ELISA and TGFbeta expression in lung tissue by immunohistochemistry was reduced on day 14 in the montelukast group. The results of this study show that montelukast inhibits the inflammatory process and development of bleomycin-induced pulmonary fibrosis in mice and that these effects may be associated with a decrease in TGFbeta expression. They also suggest that montelukast may serve as a new therapy for patients with interstitial pulmonary fibrosis. PMID- 17397876 TI - Serotonin enhances beta-endorphin secretion to lower plasma glucose in streptozotocin-induced diabetic rats. AB - Although serotonin, serotonin uptake inhibitors and serotonin precursors (including tryptophan or 5-hydroxytryptophan) are known to have hypoglycemic action in rodents or human, it is not clear whether serotonin has hypoglycemic effect in streptozotocin-induced diabetic rats (STZ-diabetic rats). The aim of this study was to investigate the action of serotonin in regulating the plasma glucose STZ-diabetic rats. Plasma glucose, insulin, beta-endorphin and adrenaline were assessed after intraperitoneal administration of serotonin. Serotonin produced hypoglycemic effects without altering plasma insulin and adrenaline levels but increasing beta-endorphin level in STZ-diabetic rats. The glycogen content in soleus muscle was increased at 90 min after application of serotonin (0.3 mg/kg) in STZ-diabetic rats. Dihydroergotamine (non-selective 5-HT receptor blocker) and pimozide (5-HT(7) receptor blocker) abolished the hypoglycemic effect of serotonin in STZ-diabetic rats. Serotonin-induced hypoglycemic effect in association with the increase of beta-endorphin release was abolished in bilaterally adrenalectomized STZ-diabetic rats. In isolated adrenal gland of STZ diabetic rats, the increase of beta-endorphin secretion in response to serotonin was reduced by either dihydroergotamine or pimozide. Pretreatment with naloxone (1.0 mg/kg, i.p.) prevented serotonin-induced plasma glucose lowering effect in STZ-diabetic rats. The results demonstrated that serotonin may activate 5-HT(7) receptor on rat adrenal gland to enhance of beta-endorphin secretion, which then stimulates the opioid receptor to increase peripheral glucose utilization, resulting in decreased plasma glucose levels in STZ-diabetic rats. PMID- 17397877 TI - Escherichia coli DNA polymerase IV contributes to spontaneous mutagenesis at coding sequences but not microsatellite alleles. AB - Slipped strand mispairing during DNA synthesis is one proposed mechanism for microsatellite or short tandem repeat (STR) mutation. However, the DNA polymerase(s) responsible for STR mutagenesis have not been determined. In this study, we investigated the effect of the Escherichia colidinB gene product (Pol IV) on mononucleotide and dinucleotide repeat stability, using an HSV-tk gene episomal reporter system for microsatellite mutations. For the control vector (HSV-tk gene only) we observed a statistically significant 3.5-fold lower median mutation frequency in dinB(-) than dinB(+) cells (p<0.001, Wilcoxon Mann Whitney Test). For vectors containing an in-frame mononucleotide allele ([G/C](10)) or either of two dinucleotide alleles ([GT/CA](10) and [TC/AG](11)) we observed no statistically significant difference in the overall HSV-tk mutation frequency observed between dinB(+) and dinB(-) strains. To determine if a mutational bias exists for mutations made by Pol IV, mutational spectra were generated for each STR vector and strain. No statistically significant differences between strains were observed for either the proportion of mutational events at the STR or STR specificity among the three vectors. However, the specificity of mutational events at the STR alleles in each strain varied in a statistically significant manner as a consequence of microsatellite sequence. Our results indicate that while Pol IV contributes to spontaneous mutations within the HSV-tk coding sequence, Pol IV does not play a significant role in spontaneous mutagenesis at [G/C](10), [GT/CA](10), or [TC/AG](11) microsatellite alleles. Our data demonstrate that in a wild type genetic background, the major factor influencing microsatellite mutagenesis is the allelic sequence composition. PMID- 17397878 TI - Cytogenetic study of the induction mechanism of chromosome-type aberrations by 1 beta-D-arabinofuranosylcytosine. AB - Treatment of human lymphocytes in the G(1) phase with 1-beta-D arabinofuranosylcytosine (AraC) results in the formation of chromosome-type aberrations, e.g., dicentric or ring chromosomes. Generally, it is accepted that DNA double-strand breaks (DSBs) cause chromosome-type aberrations. However, AraC lacks the functional groups necessary to induce such breaks, suggesting that it does not induce DSBs directly. In this study, we show that induction of DSBs is not mediated by direct action by AraC on DNA. Second, we demonstrate that the induction of DSBs or chromosome-type aberrations by AraC in human lymphocytes is inhibited by cycloheximide (CHM), an inhibitor of protein synthesis. These data suggest that newly synthesized proteins in AraC-treated lymphocytes mediate the induction of the DSBs. Thus, we suggest that AraC-induced endonucleolytic protein(s) in lymphocytes mediate the formation of chromosome-type aberrations. PMID- 17397879 TI - DNA damage, mutagenesis and cardiovascular disease. PMID- 17397880 TI - Existence of a threshold-like dose for gamma-ray induction of thymic lymphomas and no susceptibility to radiation-induced solid tumors in SCID mice. AB - Severe combined immune deficiency (SCID) mice exhibit limited repair of DNA double-strand breaks and are sensitive to ionizing radiation due to a mutation of the DNA-dependent protein kinase catalytic subunit gene. To elucidate the effects of deficient DNA double-strand break repair on radiation-induced carcinogenesis, the dose-response relationship for the induction of all tumor types was examined in wild-type and SCID mice. In wild-type mice, the incidence of thymic lymphomas at gamma-ray doses up to 1 Gy was almost equal to the background level, increased gradually above 1 Gy, and reached a maximum of 12.5% at 5 Gy, which is indicative of a threshold dose of less than 1 Gy. SCID mice were extremely susceptible to the induction of spontaneous and radiation-induced thymic lymphomas. The incidence of thymic lymphomas in SCID mice irradiated with 0.1 Gy or less was similar to the background level; that is, it increased markedly from 31.7% at 0.1 Gy to 51.4% at 0.25 Gy, and reached a maximum of 80.6% at 2 Gy, suggesting the presence of a threshold-like dose at low gamma-ray doses, even in radiosensitive SCID mice. As the average latency for the induction of thymic lymphomas at 0.1 Gy was significantly shortened, the effect of 0.1 Gy gamma-rays on thymic lymphoma induction was marginal. The high susceptibility of SCID mice to develop thymic lymphomas indicates that thymic lymphomas are induced by a defect in DNA double strand break repair or V(D)J recombination. Excessive development of tumors other than thymic and nonthymic lymphomas was not observed in SCID mice. Furthermore, our data suggest that the defective double-strand break repair in SCID mice is not a major determinant for the induction of nonlymphoid tumors. PMID- 17397881 TI - Substance P provides neuroprotection in cerebellar granule cells through Akt and MAPK/Erk activation: evidence for the involvement of the delayed rectifier potassium current. AB - In the current study, we have evaluated the ability of substance P (SP) and other neurokinin 1 receptor (NK1) agonists to protect, in a dose- and time-dependent manner, primary cultures of rat cerebellar granule cells (CGCs) from serum and potassium deprivation-induced cell death (S-K5). We also established the presence of SP high affinity NK1 transcripts and the NK1 protein localization in the membrane of a sub-population of CGCs. Moreover, SP significantly and dose dependently reduced the Akt 1/2 and Erk1/2 dephosphorylation induced by S-K5 conditions, as demonstrated by Western blot analysis. Surprisingly, in SP-treated CGCs caspase-3 activity was not inhibited, while the calpain-1 activity was moderately reduced. Corroborating this result, SP blocked calpain-mediated cleavage of tau protein, as demonstrated by the reduced appearance of a diagnostic fragment of 17 kDa by Western blot analysis. In addition, SP induced a significant reduction of the delayed rectifier K+ currents (Ik) in about 42% of the patched neurons, when these were evoked with depolarizing potential steps. Taken together, the present results demonstrate that the activation of NK1 receptors expressed in CGCs promote the neuronal survival via pathways involving Akt and Erk activation and by inhibition of Ik which can contribute to the neuroprotective effect of the peptide. PMID- 17397883 TI - Steviol glycoside biosynthesis. AB - Steviol glycosides are found in high concentrations in the leaves of the Paraguayan perennial herb Stevia rebaudiana and their intense sweetness, as well as high concentration in Stevia leaf tissue, has made them the subject of research interest for over 100 years. Steviol glycosides are diterpenoids whose biosynthetic pathways share four steps in common with gibberellic acid formation. The convergence of genomics and plant biochemistry has led to the rapid elucidation of the genes coding for the various enzymes in the biosynthetic pathway. Functional characterization of the enzymes coded for by those genes is on-going. The first committed step in the pathway is the synthesis of the aglycone steviol and the various glycosides found in the leaf tissue result from the elaboration of steviol by a number of glucosyltransferases. PMID- 17397882 TI - Orienting auditory spatial attention engages frontal eye fields and medial occipital cortex in congenitally blind humans. AB - What happens in vision-related cortical areas when congenitally blind (CB) individuals orient attention to spatial locations? Previous neuroimaging of sighted individuals has found overlapping activation in a network of frontoparietal areas including frontal eye fields (FEF), during both overt (with eye movement) and covert (without eye movement) shifts of spatial attention. Since voluntary eye movement planning seems irrelevant in CB, their FEF neurons should be recruited for alternative functions if their attentional role in sighted individuals is only due to eye movement planning. Recent neuroimaging of the blind has also reported activation in medial occipital areas, normally associated with visual processing, during a diverse set of non-visual tasks, but their response to attentional shifts remains poorly understood. Here, we used event-related fMRI to explore FEF and medial occipital areas in CB individuals and sighted controls with eyes closed (SC) performing a covert attention orienting task with endogenous verbal cues and spatialized auditory targets. We found robust stimulus-locked FEF activation of all CB subjects, similar to and stronger than in SC, suggesting that FEF plays a role in endogenous orienting of covert spatial attention even in individuals in whom voluntary eye movements are irrelevant. We also found robust activation in bilateral medial occipital cortex in CB but not in SC subjects. The response decreased below baseline following endogenous verbal cues but increased following auditory targets, suggesting that the medial occipital area in CB does not directly engage during cued orienting of attention but may be recruited for processing of spatialized auditory targets. PMID- 17397884 TI - Activity-guided isolation of antiplasmodial dihydrochalcones and flavanones from Piper hostmannianum var. berbicense. AB - The bioassay-guided purification of an n-hexane extract from the leaves of Piper hostmannianum var. berbicense led to the isolation of four monoterpene or prenyl substituted dihydrochalcones (1a, 1b, 2, 3) as well as the known compounds 2',6' dihydroxy-4'-methoxydihydrochalcone (4), linderatone (5), strobopinin (6), adunctin E (7) and (-)-methyllinderatin (8). Their structures were established on the basis of NMR and X-ray analysis. (-)-Methyllinderatin, linderatone and 2',6' dihydroxy-4'-methoxydihydrochalcone exhibited the most potent antiplasmodial activity with IC50 values of 5.64, 10.33 and 12.69 microM, respectively against both chloroquine-sensitive and resistant strains of Plasmodium falciparum (F32,FcB1). The activity of (-)-methyllinderatin was confirmed in vivo against Plasmodium vinckei petteri in mice (80% of reduction of parasitemia) at a dose of 20 mg/kg/day. PMID- 17397885 TI - The effects of increased cAMP content on inflammation, oxidative stress and PDE4 transcripts during Brucella melitensis infection. AB - Cyclic AMP (cAMP) is a key intracellular second messenger which at increased levels has been shown to have anti-inflammatory and tissue-protective effects. Its concentration is determined by the activities of both adenylate cyclase (AC) and the phosphodiesterase (PDE) enzymes. The aim of this study was to compare the effects of increased cAMP and glucocorticoid dexamethasone administration on B. melitensis-induced lipid peroxidation, Brucella suppressed antioxidant enzyme activities and PDE4 transcripts in rats. Intracellular cyclic AMP level was elevated by two different approaches; activation of AC and inhibition of PDE activities. Rats were inoculated with B. melitensis for seven days then a single dose of nonselective PDE inhibitor 3-isobutyl-1-methylxanthine (IBMX), the adenylate cyclase activator forskolin and dexamethasone were administrated to each infected group, and animals were challenged for 48 h. Brucella-induced lipid peroxidation was significantly reduced by the cAMP elevating agents as well as dexamethasone administration in plasma, liver and spleen. The antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) activities were significantly decreased by the pathogen. Whilst suppressed GSH-Px activity was reversed by cAMP elevating agents, SOD activity was not restored. Superoxide generating enzyme xanthine oxidase activity was not altered at the end of the infection period. Brucella infection increased plasma IL-12 level and this effect was also suppressed by the cAMP elevating agents, whereas TNF-alpha, IFN-gamma and IL-10 levels were unchanged. Intracellular cAMP levels are entirely hydrolyzed by cAMP-specific PDE 4 isozymes (PDE4s) in inflammatory and immunocompetent cells. Brucella reduced mRNA transcript levels for PDE4A by 40%, though PDE4B and 4D transcriptions were being unaffected in spleen. It was concluded that B. melitensis infection decreased activity of the antioxidant defence system, induced lipid peroxidation and suppressed PDE4A transcription. Administration of cAMP elevating agents exhibited similar affect with dexamethasone on lipid peroxidation, IL-12 production and antioxidant enzyme activities in Brucella infection. PMID- 17397886 TI - [Evulsion of the optic nerve, a diagnosis to be recognised!]. AB - INTRODUCTION: Evulsion of the optic nerve is a rare but serious injury. It occurs generally after an ocular contusion and may cause blindness. CASE: We report the case of a 53-year-old woman who presented with a blunt ocular trauma after having been punched by her husband. Initial examination of the left eye was impossible due to a major palpebral oedema. A CT scan of the orbit revealed a thickened optic nerve. No improvement was noted. DISCUSSION: Optic nerve avulsion is often caused by sudden and forceful rotation of the eye with tearing of the optic nerve as its globe entry level. The diagnosis can be confirmed by examination of the ocular fundus or by medical imaging such as CT scan of the orbital cavity as in our case report. The prognosis is usually poor. PMID- 17397887 TI - Estradiol affects liver mitochondrial function in ovariectomized and tamoxifen treated ovariectomized female rats. AB - Given the tremendous importance of mitochondria to basic cellular functions as well as the critical role of mitochondrial impairment in a vast number of disorders, a compelling question is whether 17beta-estradiol (E2) modulates mitochondrial function. To answer this question we exposed isolated liver mitochondria to E2. Three groups of rat females were used: control, ovariectomized and ovariectomized treated with tamoxifen. Tamoxifen has antiestrogenic effects in the breast tissue and is the standard endocrine treatment for women with breast cancer. However, under certain circumstances and in certain tissues, tamoxifen can also exert estrogenic agonist properties. We observed that at basal conditions, ovariectomy and tamoxifen treatment do not induce any statistical alteration in oxidative phosphorylation system and respiratory chain parameters. Furthermore, tamoxifen treatment increases the capacity of mitochondria to accumulate Ca(2+) delaying the opening of the permeability transition pore. The presence of 25 microM E2 impairs respiration and oxidative phosphorylation system these effects being similar in all groups of animals studied. Curiously, E2 protects against lipid peroxidation and increases the production of H(2)O(2) in energized mitochondria of control females. Our results indicate that E2 has in general deleterious effects that lead to mitochondrial impairment. Since mitochondrial dysfunction is a triggering event of cell degeneration and death, the use of exogenous E2 must be carefully considered. PMID- 17397889 TI - Molecular processes in cellular arsenic metabolism. AB - Elucidating molecular processes that underlie accumulation, metabolism and binding of iAs and its methylated metabolites provides a basis for understanding the modes of action by which iAs acts as a toxin and a carcinogen. One approach to this problem is to construct a conceptual model that incorporates available information on molecular processes involved in the influx, metabolism, binding and efflux of arsenicals in cells. This conceptual model is initially conceived as a non-quantitative representation of critical molecular processes that can be used as a framework for experimental design and prediction. However, with refinement and incorporation of additional data, the conceptual model can be expressed in mathematical terms and should be useful for quantitative estimates of the kinetic and dynamic behavior of iAs and its methylated metabolites in cells. Development of a quantitative model will be facilitated by the availability of tools and techniques to manipulate molecular processes underlying transport of arsenicals across cell membranes or expression and activity of enzymes involved in methylation of arsenicals. This model of cellular metabolism might be integrated into more complex pharmacokinetic models for systemic metabolism of iAs and its methylated metabolites. It may also be useful in development of biologically based dose-response models describing the toxic and carcinogenic actions of arsenicals. PMID- 17397888 TI - Photodynamic therapy with the phthalocyanine photosensitizer Pc 4: the case experience with preclinical mechanistic and early clinical-translational studies. AB - Photodynamic therapy (PDT) is emerging as a promising non-invasive treatment for cancers. PDT involves either local or systemic administration of a photosensitizing drug, which preferentially localizes within the tumor, followed by illumination of the involved organ with light, usually from a laser source. Here, we provide a selective overview of our experience with PDT at Case Western Reserve University, specifically with the silicon phthalocyanine photosensitizer Pc 4. We first review our in vitro studies evaluating the mechanism of cell killing by Pc 4-PDT. Then we briefly describe our clinical experience in a Phase I trial of Pc 4-PDT and our preliminary translational studies evaluating the mechanisms behind tumor responses. Preclinical work identified (a) cardiolipin and the anti-apoptotic proteins Bcl-2 and Bcl-xL as targets of Pc 4-PDT, (b) the intrinsic pathway of apoptosis, with the key participation of caspase-3, as a central response of many human cancer cells to Pc 4-PDT, (c) signaling pathways that could modify apoptosis, and (d) a formulation by which Pc 4 could be applied topically to human skin and penetrate at least through the basal layer of the epidermis. Clinical-translational studies enabled us to develop an immunohistochemical assay for caspase-3 activation, using biopsies from patients treated with topical Pc 4 in a Phase I PDT trial for cutaneous T-cell lymphoma. Results suggest that this assay may be used as an early biomarker of clinical response. PMID- 17397891 TI - DPSM technique for ultrasonic field modelling near fluid-solid interface. AB - Distributed point source method (DPSM) is gradually gaining popularity in the field of non-destructive evaluation (NDE). DPSM is a semi-analytical technique that can be used to calculate the ultrasonic fields produced by transducers of finite dimension placed in homogeneous or non-homogeneous media. This technique has been already used to model ultrasonic fields in homogeneous and multi-layered fluid structures. In this paper the method is extended to model the ultrasonic fields generated in both fluid and solid media near a fluid-solid interface when the transducer is placed in the fluid half-space near the interface. Most results in this paper are generated by the newly developed DPSM technique that requires matrix inversion. This technique is identified as the matrix inversion based DPSM technique. Some of these results are compared with the results produced by the Rayleigh-Sommerfield integral based DPSM technique. Theory behind both matrix inversion based and Rayleigh-Sommerfield integral based DPSM techniques is presented in this paper. The matrix inversion based DPSM technique is found to be very efficient for computing the ultrasonic field in non-homogeneous materials. One objective of this study is to model ultrasonic fields in both solids and fluids generated by the leaky Rayleigh wave when finite size transducers are inclined at Rayleigh critical angles. This phenomenon has been correctly modelled by the technique. It should be mentioned here that techniques based on paraxial assumptions fail to model the critical reflection phenomenon. Other advantages of the DPSM technique compared to the currently available techniques for transducer radiation modelling are discussed in the paper under Introduction. PMID- 17397890 TI - 4-Methoxyestradiol-induced oxidative injuries in human lung epithelial cells. AB - Epidemiological studies indicated that people exposed to dioxins were prone to the development of lung diseases including lung cancer. Animal studies demonstrated that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) increased liver tumors and promoted lung metaplasia in females. Metabolic changes in 17beta estradiol (E(2)) resulted from an interaction between TCDD and E(2) could be associated with gender difference. Previously, we reported that methoxylestradiols (MeOE(2)), especially 4-MeOE(2), accumulated in human lung cells (BEAS-2B) co-treated with TCDD and E(2). In the present study, we demonstrate unique accumulation of 4-MeOE(2), as a result of TCDD/E(2) interaction and revealed its bioactivity in human lung epithelial cell line (H1355). 4-Methoxyestradiol treatment significantly decreased cell growth and increased mitotic index. Elevation of ROS and SOD activity, with a concomitant decrease in the intracellular GSH/GSSG ratio, was also detected in 4-MeOE(2) treated cells. Quantitative comet assay showed increased oxidative DNA damage in the 4-MeOE(2)-treated H1355 cells, which could be significantly reduced by the anti-oxidant N-acetylcysteine (NAC). However, inhibition of cell growth and increase in mitotic arrest induced by 4-MeOE(2) were unaffected by NAC. We concluded that 4-MeOE(2) accumulation resulting from TCDD and E(2) interaction would contribute to the higher vulnerability on lung pathogenesis in females when exposed to TCDD. PMID- 17397892 TI - Differences in tropism and pH dependence for glycoproteins from the Clade B1 arenaviruses: implications for receptor usage and pathogenicity. AB - The Clade B lineage of the New World arenaviruses contains four viruses capable of causing severe hemorrhagic fevers in humans. Within this group, the B1 sub lineage contains the pathogenic viruses Junin (JUNV) and Machupo (MACV), as well as the non-pathogenic Tacaribe virus (TCRV). In order to elucidate differences that may determine pathogenicity, we studied the entry pathways directed by the glycoproteins (GPs) from these related B1 viruses, using pseudotyped retroviral vectors and GP1 immunoadhesin constructs. Our data revealed variations in the efficiency with which different cell types could be transduced by B1 vectors, and this correlated with the ability of the immunoadhesins to bind to those cells. Interestingly, the tropism directed by the TCRV GP proved to be distinct from that of JUNV and MACV, in particular on lymphocyte cell lines. In addition, the GPs showed variations in their sensitivity to an inhibitor of endosome acidification, with the TCRV GP again being the outlier. Together these data suggest that more than one entry pathway can be used by these closely related viruses and that the ability to cause human disease may be highly dependent on receptor usage. PMID- 17397893 TI - HLA homozygosity does not adversely affect measles vaccine-induced cytokine responses. AB - The association between HLA homozygosity and measles-specific Th(1) (IFN-gamma, IL-2 and IL-12p40) and Th(2) (IL-4 and IL-10) cytokine responses were assessed in a group of 339 healthy schoolchildren 12-18 years of age previously immunized with two doses of live-attenuated measles virus vaccine. No associations were observed between class I HLA homozygosity and measles-specific cytokine levels. Children who were homozygous at the class II DRB1, DQA1, DPA1 and DPB1 loci had higher median IFN-gamma secretion levels compared with children who were heterozygous for DRB1 (77.7 vs. 39.5 pg/ml, p=0.05), DQA1 (60.9 vs. 36.6 pg/ml, p=0.03), DPA1 (46.1 vs. 27.1 pg/ml, p=0.01) and DPB1 (61.5 vs. 36.0 pg/ml, p=0.01) loci, respectively. Homozygosity at increasing numbers of HLA loci ( >or=4) was associated with increased IFN-gamma secretion levels (test for trend p value=0.01). Our results suggest that HLA homozygosity showed no disadvantage for measles-specific cytokine responses and instead was associated with increased IFN gamma levels. PMID- 17397894 TI - The role of lysine 186 in HIV-1 integrase multimerization. AB - HIV-1 integrase (IN) catalyzes biochemical reactions required for viral cDNA insertion into host cell chromosomal DNA, an essential step in the HIV-1 replication cycle. In one of these reactions, the two ends of the linear viral cDNA are believed to be simultaneously ligated to chromosomal DNA by a tetrameric form of IN. The structure of the full-length IN tetramer is not known but a model consisting of the N-terminal domain and the catalytic core revealed basic residues 186 to 188 at the interface between the two IN dimers. We found that alteration of these residues, in particular changing IN lysine residue 186 to glutamate (K186Q), impairs IN oligomerization in the yeast two-hybrid system and decreases oligomeric forms of IN within virions. When expressed independently of other viral proteins in human cells, IN-K186Q did not concentrate in the nucleus as did wild-type IN. Co-expression of wild-type IN restored the multimerization defects of IN-K186Q, in both the two-hybrid system and in virions, and also rescued the nuclear targeting defects. Virions bearing IN-K186Q were not infectious in a single cycle of replication but when mixed virions containing two different IN mutants were produced, IN-K186Q was capable of complementing the catalytically inactive mutant IN-D116A. Our biochemical and functional data support the crystallographic model in which IN residue K186 lies at the interface between IN dimers and suggest that tetramerization is important, not only for concerted integration, but also for IN nuclear targeting. PMID- 17397895 TI - Genetic analysis of J-virus and Beilong virus using minireplicons. AB - J-virus (JPV), isolated from wild mice in Australia, and Beilong virus (BeiPV), originally isolated from human mesangial cells in China and subsequently detected in rat mesangial cells, represent a new group of paramyxoviruses which have exceptionally large genomes (>19 kb) and contain more than six transcriptional units. In this study, minireplicons were employed to assess the taxonomic status of JPV and BeiPV. Our results demonstrated that, whilst the genome replication machineries of JPV and BeiPV can be interchanged, they were not functional when exchanged with that of Nipah virus. These studies indicate that JPV and BeiPV are closely related to each other and support the classification of these two viruses into a separate genus. In addition, the minireplicons were also used to demonstrate that these large-genome viruses also comply with the 'rule of six' and that over-expression of the C protein has a detrimental effect on minigenome replication. PMID- 17397896 TI - Infant color vision: infants' spontaneous color preferences are well behaved. AB - Stochastic transitivity (ST) is a property of preferences among pairs of objects formed from three alternatives, a, b, and c. In general, ST states that if a is preferred to b, and b is preferred to c, then a will be preferred to c. Stochastic transitivity can be weak, moderate, strong or strict (see text). In the present paper, we analyse the presence and degree of ST in the data from two experiments concerning 12-week-old infants' spontaneous color preferences. In the first experiment (Triads), we tested five sets of three stimuli in pairs of two (a vs. b, b vs. c, a vs. c). In each case two stimuli were chromatic and one was White. Strict ST was seen in all cases. In Experiment 2 (Complementaries), we tested White against pairs of stimuli from opposite sides of the White point (red vs. blue-green, blue vs. yellow, and green vs. purple). The purities required for equal (50/50) preference between the two chromatic stimuli were consistent with the preferences for each of the two stimuli over White. In addition, 12 new triads were generated from the Complementaries experiment. Strict ST was seen in six out of 12 cases, and Moderate ST was seen in the other six. As discussed further in the accompanying paper [Zemach, I. K., Chang, S., & Teller, D. Y. (2007). Infant color vision: prediction of infants' spontaneous color preferences], White was the least preferred stimulus in every triad tested. Although more extensive studies are needed, the data suggest that infants' hue preferences are reasonably well behaved across different choices of stimulus pairs. PMID- 17397897 TI - Release of polycyclic aromatic compounds into a Mediterranean creek (Catalonia, NE Spain) after a forest fire. AB - Wildfires produce polycyclic aromatic compounds (PACs), among which polycyclic aromatic hydrocarbons (PAHs) are of environmental concern. After a fire affecting a Mediterranean creek, we studied the distribution and short-term permanence of PACs and of 16 priority PAH pollutants in the aquatic environment. PACs were traced through absorbance and fluorescence emissions and organic carbon concentrations. PAHs were analysed by gas chromatography. High TOC/DOC concentrations and fluorescence/absorbance values reflected abnormal inputs of fire-induced aromatic compounds. Their concentrations decreased gradually but showed sporadic peaks after precipitation. Fifteen months later, values still surpassed background levels. Initially concentrations of individual and total PAH species of the dissolved, particulate and sediment phases were increased. Then they fluctuated with new PAH inputs related with runoff, leaching and soil erosion following precipitation events. All PAH concentrations approached background levels 15 months after the fire, and never reached values of toxicological concern. However, some bioaccumulation may have occurred. PMID- 17397898 TI - Comments on: "Removal of the surfactant sodium dodecylbenzene sulphonate from water by simultaneous use of ozone and activated carbon". PMID- 17397899 TI - Spatial distribution and physiological state of bacteria in a sand column experiment during the biodegradation of toluene. AB - Toxic organic contaminants frequently serve as growth substrates for bacteria. However, long-term exposure to the organic contaminants can result in significant stress or "injury" to bacterial cells such that bacteria may lose, either temporarily or permanently, their capacity to degrade a specific toxic organic contaminant. In order to understand the relationship between biodegradability and physiological conditions of bacteria after a prolonged exposure to a contaminant, biomass samples collected from a sand column experiment, with toluene as the carbon source, were analyzed for bacterial physiology and spatial population distribution in the porous media. The column was seeded with three bacterial isolates that perform aerobic (Pseudomonas putida F1), denitrifying (Thauera aromatica T1), and facultative (Ralstonia pickettii PKO1) degradation of toluene were analyzed. Total, viable but not culturable with toluene, and toluene culturable cells were enumerated using 4'6-diamidino-2-phenylindole (DAPI) staining and plate counting methods. Comparison of three types of cell counts showed that toluene-culturable cells were less than 40% of the total cell numbers. However, viable colonies transferred to a toluene media after cultivation on rich media regained their ability to degrade toluene. This implies that the temporary loss of their toluene degradation capacity is either due to an intracellular accumulation of degradation by-products, which have to be consumed in order for the cells to degrade toluene, or it is possible that cells have shifted to degrade other substrates such as toluene degradation intermediates or organic materials resulting from cell turnover. Comparison of cell counts with toluene concentration showed no exponential increase in total and viable cell numbers, as reported for flat bed biofilm reactor experiments. The overall fraction of toluene-culturable cells was highest at the highest toluene concentration near the column inlet, which indicates that the observed temporary loss of toluene culturability was not solely caused by a direct toxic effect from the long-term exposure to toluene. PMID- 17397900 TI - Comparisons of polychromatic and monochromatic UV-based treatments of bisphenol-A in water via toxicity assessments. AB - Polychromatic ultraviolet irradiation, such as from medium pressure (MP) Hg lamps may enhance the UV degradation of environmental pollutants as compared to low pressure (LP) Hg UV sources emitting monochromatic irradiation. Typically, studies involving destruction of environmental pollutants such as endocrine disrupting compounds (EDCs) are based on measurement of the parent compound decay using analytical chemistry, but such information is insufficient to determine an effective treatment endpoint because the identity and biological activity of many transformation products remain unknown. Bioanalytical methods to assess residual biological activity of a treated water offers one means to compare removal efficiency of EDC activity between MP- and LP-UV lamps under photolysis and UV/H2O2 oxidation. In this study, changes in estrogenic activity of bisphenol-A (BPA) as a function of UV treatment were evaluated using both an in vitro yeast estrogen screen and in vivo vitellogenin assay with Japanese medaka (Oryzias latipes) fish. Decay of BPA parent compound and formation of degradation products were followed using HPLC analysis. Results demonstrated that MP-UV direct photolysis more effectively removed BPA and associated estrogenic activity compared to LP-UV lamps. UV in combination with H2O2 significantly removed estrogenic activity in vitro and in vivo compared to direct photolysis; however, no significant difference in removal rates was found between the two lamps under UV/H2O2 oxidation. Furthermore, the UV/H2O2 process was effective for reducing embryo toxicity of BPA, but resulted in the production of acidic intermediates, causing acute toxicity and delayed hatching in some medaka embryos. PMID- 17397901 TI - A novel source for dioxins present in recycled fat from gelatin production. AB - Within a survey on dioxins in animal fat used as feed ingredient, a sample originating from pigs offal was shown to contain 50 ng Toxic Equivalents (TEQ) PCDD/PCDFs kg(-1) fat. Further investigation revealed fat samples with levels as high as 440 ng TEQ kg(-1) fat and contaminated feed with a highest level of 8.4 ng TEQ kg(-1) feed. The congener pattern was dominated by 1,2,3,7,8-PeCDD and 2,3,7,8-TCDD, and was not recognized from any previous incident or known dioxin source. Remarkably, 2,3,7,8-substituted congeners were much more abundant than their non-2,3,7,8-substituted counterparts. The sampled fat was derived from a gelatin production plant. Broken filters, used to clean the hydrochloric acid (HCl) used in the process, caused the dioxin contamination. The fat was primarily used for pig feed. A new physiologically-based pharmacokinetic (PBPK) model for lipophilic contaminants in growing slaughter pigs predicted levels at slaughter varying between 40 pg TEQ g(-1) fat (worst-case) and 2.5-7pgTEQ g(-1) fat under more realistic scenarios. Almost 300 farms were temporarily blocked. Many fat samples of pigs were analyzed using a combined approach of DR CALUX and GC/HRMS. Levels in contaminated pig fat were around the EU-limit of 1 pg TEQ g(-1) fat, with some samples up to 2-3 pg TEQ g(-1) fat. Of 80 negative samples analyzed by DR CALUX and GC/HRMS no false-negatives were obtained, whereas 36 and 62 of the 80 samples classified suspected with the bioassay had GC/HRMS levels above respectively the tolerance and action limits. It is concluded that novel and unexpected dioxin sources remain a threat to the food chain and require the proper evaluation and monitoring of production processes, including chemicals used therein. PMID- 17397902 TI - Flow cytometry assessment of cytotoxicity and reactive oxygen species generation by single and binary mixtures of cadmium, zinc and copper on populations of the ciliated protozoan Tetrahymena thermophila. AB - In the present study, we have assessed by flow cytometry, the cytotoxicity of the heavy metals Cd, Zn and Cu on populations of the ciliated protozoan Tetrahymena thermophila. The obtained LC(50) for these metals was estimated as 0.195mgCdl( 1), 3.58mgZnl(-1) and 0.47mgCul(-1), respectively. As a result, the toxicity rank for this eukaryotic microorganism is Cd>Cu>>Zn. Using the same methodology and the Concentration Addition approach, the toxicity of binary mixtures of these metals was evaluated in order to detect the type of interaction between these metals. Results indicated that antagonism is the predominant interaction but it can change to additivity or even to synergism at high metal concentrations. Besides, the concentration ratio between metals plays also a crucial role in determining the type of metallic interaction, at least in Tetrahymena. Cytotoxicity data from single and bimetallic mixtures have been compared with those from selected microalgae, other species of ciliates, fish and mammalian cell lines and metazoan. By other hand, we have detected the mitochondrial generation of peroxides induced by both single and binary treatments with Cd, Zn and Cu on populations of Tetrahymena, using the specific fluorophore dyhidrorhodamine. The nature and concentrations of metal as well as the metallic ratio were important factors in reactive oxygen species production. All results found in T. thermophila are compared with previous reports in other organisms and, some explanations and hypothesis to support results are given, including the involvement of metallothioneins as antioxidants and their role in the binding of metal cations. PMID- 17397903 TI - CuFe2O4/activated carbon composite: a novel magnetic adsorbent for the removal of acid orange II and catalytic regeneration. AB - CuFe2O4/activated carbon magnetic adsorbents, which combined the adsorption features of activated carbon with the magnetic and the excellent catalytic properties of powdered CuFe2O4, were developed using a simple chemical coprecipitation procedure. The prepared magnetic composites can be used to adsorb acid orange II (AO7) in water and subsequently, easily be separated from the medium by a magnetic technique. CuFe2O4/activated carbon magnetic adsorbents with mass ratio of 1:1, 1:1.5 and 1:2 were prepared. Magnetization measurements, BET surface area measurements, powder XRD and SEM were used to characterize the prepared adsorbents. The results indicate that the magnetic phase present is spinel copper ferrite and the presence of CuFe2O4 did not significantly affect the surface area and pore structure of the activated carbon. The adsorption kinetics and adsorption isotherm of acid orange II (AO7) onto the composites at pH 5.2 also showed that the presence of CuFe2O4 did not affect the adsorption capacity of the activated carbon. The thermal decomposition of AO7 adsorbed on the activated carbon and the composite was investigated by in situ FTIR, respectively. The results suggest that the composite has much higher catalytic activity than that of activated carbon, attributed to the presence of CuFe2O4. The variation of the adsorption capacity of the composites after several adsorption-regeneration cycles has also been studied. PMID- 17397904 TI - Nonylphenol algal bioaccumulation and its effect through the trophic chain. AB - Nonylphenol is a metabolic intermediate from the microbial transformation of detergents used worldwide. While nonylphenol shows some acute toxicity, it is also able to mimic important hormones resulting in the disruption of several processes by interfering with the signals that control the overall physiology of the organism. The effect of the pollutant nonylphenol (NP) through the trophic chain was studied. Microalgae Isochrysis galbana was able to bioconcentrate NP 6940 times, where 77% of initial NP (100microgl(-1)) is accumulated intracellularly after 1-h incubation. Crustacean Artemia fransiscana showed 25% higher growth when fed with NP-rich algae. However, Artemia metabolized almost all NP ingested and only traces of NP could be found in the organism, eliminating future NP effects. Zebrafish (Brachydanio rerio) were affected by the presence of 171microgg(-1) of NP in the diet, showing higher levels of the hormone vitellogenin and lower levels of cytochrome P450 activity. These results showed that organisms placed in the first level of trophic chain are able to significantly bioconcentrate the pollutant and endocrine disruptor NP. These grassed organisms affect the growth of crustacean. Moreover, the organisms placed on the top of some trophic chains, such as fish, could be affected by the presence of NP in their food, in both the hormone levels and metabolic enzymes. This work shows that the environmental presence of NP should be considered as a risk for the organisms living in an ecosystem. PMID- 17397905 TI - Histopathologic assessment of tumor regression after neoadjuvant chemotherapy in advanced-stage ovarian cancer. AB - To date, no histopathologic criteria have been established to describe treatment response after neoadjuvant chemotherapy in ovarian cancer. The aim of this study was to identify histopathologic features of tumor regression in ovarian cancer specimens obtained after neoadjuvant chemotherapy regarding their ability to indicate treatment response. This study systematically evaluated histopathologic features of tumor regression in advanced-stage ovarian cancer treated with neoadjuvant chemotherapy (n = 49) and in a control group treated with primary surgery (n = 35). In addition, the largest tumor size was measured in the surgical specimens. Overall survival served as the reference standard with a median follow-up of 49 months. There was a significantly higher presence of regressive changes in the postchemotherapy group compared with the untreated control group (P < or = .04). The presence of scattered solitary tumor cells, fibrosis, foamy macrophages, and giant cells of foreign-body type each indicated previous neoadjuvant chemotherapy with high specificity (80.0%-100%) but with low sensitivity (18.4%-63.3%). Inflammatory cell infiltrates, isolated psammoma bodies, and hemosiderin were also associated with previous chemotherapy but with lower specificity. The presence of necrosis was significantly correlated with larger tumor size within the specimens (rho = 0.5, P < .0001) and was more often found in the control group. For both groups, the extent of regressive changes, evaluated as a single parameter or in combination, showed no correlation with overall survival. However, patients with absence of residual tumor, scattered solitary tumor cells, or residual tumor foci of 5 mm or less after neoadjuvant chemotherapy had a significantly longer median overall survival of 45.6 versus 27.3 months in patients with larger tumors (P = .02). Various histopathologic features generally associated with posttreatment changes did not allow differentiation of responding from nonresponding patients and provided no prognostic information. The residual tumor size after neoadjuvant chemotherapy was the only criterion significantly correlated with treatment response and subsequent overall survival. PMID- 17397906 TI - A unifying concept of trophoblastic differentiation and malignancy defined by biomarker expression. AB - Several trophoblast phenotypes, including cytotrophoblast, syncytiotrophoblast, and extravillous trophoblast, emerge during gestation. To clarify the lineage relationship between these subtypes, we profiled p63 localization in developing and term placental tissue, as well as in trophoblastic tumors, using antibodies specific to full-length (TAp63) and one against all p63 isoforms (TAp63 and DeltaNp63). Localization of p63 was compared with that of biomarkers of proliferation and trophoblastic differentiation, including mib-1, inhibin, and MelCAM. In early gestation, p63 was localized principally to villous cytotrophoblast after contact with the villous mesenchyme, absent in the trophoblast columns, and early implantation trophoblast. In the maturing placenta, intraplacental perivillous fibrin correlated with the emergence of a p63-positive "transitional" (vacuolated) extravillous trophoblast from cytotrophoblast, which differentiated further into a "mature" p63-negative extravillous trophoblast. The same lineage pathway emerged from entrapped villi on the chorionic membrane. Virtually all p63 immunopositivity was attributed to dominant-negative p63. The immunophenotypic patterns seen in the immature and mature placenta permit the resolution of all trophoblastic phenotypes within 3 lineage pathways of cytotrophoblast differentiation, including cytotrophoblast-to trophoblast column/implantation site, cytotrophoblast-to-syncytiotrophoblast, and cytotrophoblast-to-mature extravillous trophoblast. In the latter pathway, a transitional (vacuolated) p63-positive extravillous trophoblast emerges from and links cytotrophoblast to mature extravillous trophoblast in intraplacental fibrin, chorionic membrane, and basal plate. The placental trophoblast is thus resolved within this continuum of differentiation. Terms such as transitional and mature extravillous trophoblast are proposed to reflect the differentiation phases of this unique epithelium. p63 staining patterns in trophoblastic tumors reflect timing of neoplastic transformation during trophoblastic differentiation. PMID- 17397908 TI - The activation of neutrophil elastase-mediated fibrinolysis is not sufficient to overcome the fibrinolytic shutdown of disseminated intravascular coagulation associated with systemic inflammation. AB - INTRODUCTION: We conducted a prospective study to test the hypothesis that the activation of neutrophil elastase-mediated fibrinolysis is insufficient to overcome the fibrinolytic shutdown of disseminated intravascular coagulation (DIC) in patients associated with systemic inflammation. MATERIALS AND METHODS: We investigated 45 consecutive patients with systemic inflammatory response syndrome (SIRS) and sepsis, classified as 11 DIC and 34 non-DIC. Fibrin degradation products by neutrophil elastase (Elastase-XDP) and by plasmin (FDP), cross-linked fibrin degradation products (D-dimer), soluble fibrin, antithrombin, protein C, plasminogen activator inhibitor-1 (PAI-1), and urinary trypsin inhibitor (UTI) were measured within 24 h after the patients met either the SIRS or sepsis criteria (day 0), as well as on days 2 and 4. RESULTS: In DIC patients, higher levels of soluble fibrin, PAI-1, and FDP and markedly lower levels of antithrombin and protein C were observed in comparison to those in non-DIC patients. DIC patients showed a significantly higher level of peak Elastase-XDP than non-DIC patients (25.7+/-5.9 vs. 16.3+/-2.6 microg/mL, respectively; p=0.0333). However, we could not find any substantial difference in the levels of Elastase-XDP, UTI, and D-dimer on each day during the study period between the two groups. Good correlations were observed between the levels of D-dimer and Elastase-XDP in both patients with and without DIC (r(s)=0.699 and r(s)=0.817, respectively), but the coefficients of determination in both groups showed low values and the slopes of the regression lines were less than 1.0. A multivariate logistic regression analysis showed the elevated peak Elastase-XDP levels to inversely correlate with death. On the other hand, the DIC patients showed a higher number of organ dysfunctions and a poorer prognosis than did the non-DIC patients. CONCLUSIONS: The activation of the neutrophil elastase-mediated fibrinolytic pathway may be insufficient to overcome the fibrinolytic shutdown by PAI-1 and may in part explain the poor prognosis of DIC patients associated with systemic inflammation. PMID- 17397909 TI - Convenience of the computed tomography perfusion method for cerebral vasospasm detection after subarachnoid hemorrhage. AB - BACKGROUND: Vasospasm is a frequent complication in the early clinical course after SAH. Although various methods have been used to measure cerebral perfusion including PET, SPECT, xenon CT, and TCD, these require the patients to remain still for a long period. In addition, TCD is operator dependent. The current study aimed to clarify the convenience of CTP for the assessment of cerebral vasospasm caused by SAH. METHODS: Nineteen patients with SAH aged 44 to 85 years (mean, 64 years) were recruited with informed consent. All patients were treated with the prevailing therapy and underwent CTP on days 6 to 9, followed by DSA and 3D-CTA to detect cerebral vasospasm. In each patient, we measured the MTT, CBF, and CBV. The reliability of CTP data was verified by comparing the data from CTP and xenon CT between the controls, and the average was calculated. Six ROIs were located symmetrically in the frontal, temporal, and occipital lobes. RESULTS: An MTT value more than 20% greater than the average indicated the progression of cerebral vasospasm, and patients with vasospasm-related infarcts exhibited an MTT more than 47% greater than the mean value (odds ratio, 50). Patients with delayed cerebral infarcts had a significantly lower mean CBF and CBV and higher MTT than patients who did not develop CI. CONCLUSION: Significant correlations between MTT and CBF values and neurovascular findings were obtained. Computed tomography perfusion can be performed in a short time and on a regular basis, and it therefore has the potential to identify cerebral vasospasm because of SAH. PMID- 17397910 TI - Prognostic factors for complete debulking in advanced ovarian cancer and its impact on survival. An exploratory analysis of a prospectively randomized phase III study of the Arbeitsgemeinschaft Gynaekologische Onkologie Ovarian Cancer Study Group (AGO-OVAR). AB - BACKGROUND: No residual tumor as result of primary surgery in advanced ovarian cancer is known as one of the most important prognostic factors. PURPOSE: To evaluate the impact of different prognostic factors for surgical outcome and to evaluate the impact of surgical outcome on survival. METHODS: Surgical data as well as survival data were documented throughout the multi-center prospective randomized phase III trial (OVAR-3) of the AGO-OVAR and were used for this exploratory analysis. In this study 798 patients with FIGO IIB-IV were first operated then randomized and homogenously treated with cisplatin/paclitaxel or carboplatin/paclitaxel. Only patients with complete surgical data (n=761) entered this analysis. RESULTS: Multivariable logistic regression analysis showed a significant decrease of probability for complete debulking without any macroscopic residual tumor for higher pre-operative tumor load (OR 0.32; 95% CI 0.17-0.61), higher FIGO stage (OR 0.22; 95% CI 0.13-0.39), worse performance status (OR 0.57; 95% CI 0.38-0.86), advanced age (OR 0.78; 95% CI 0.65-0.94) and presence of peritoneal carcinomatosis (OR 0.17; 95% CI 0.10-0.28). Surgery in centers with surgeons who performed comprehensive surgical debulking including retroperitoneal lymphadenectomy and peritoneal stripping was associated with higher rates of complete debulking compared to surgery in other centers (32.8% vs. 22.9%, p=0.007). This resulted in a markedly improved overall survival (p=0.045). This effect was held true after adjustment for prognostic factors (HR 0.77, 95% CI 0.63-0.94, p=0.012). CONCLUSION: Post-operative residual tumor is one of the most important independent prognostic factor for survival. Our results suggest an advantage for aggressive primary surgery and complete debulking. This surgical goal was achieved more often in experienced centers. PMID- 17397911 TI - An experimental and modeling-based approach to locate IgE epitopes of plant profilin allergens. AB - BACKGROUND: Plant profilins are actin-binding proteins that form a well-known panallergen family responsible for cross-sensitization between plant foods and pollens. Melon profilin, Cuc m 2, is the major allergen of this fruit. OBJECTIVE: We sought to map IgE epitopes on the 3-dimensional structure of Cuc m 2. METHODS: IgE binding to synthetic peptides spanning the full Cuc m 2 amino acid sequence was assayed by using a serum pool and individual sera from 10 patients with melon allergy with significant specific IgE levels to this allergen. Three-dimensional modeling and potential epitope location were based on analysis of both solvent exposure and electrostatic properties of the Cuc m 2 surface. RESULTS: Residues included in synthetic peptides that exerted the strongest IgE-binding capacity defined 2 major epitopes (E1, consisting of residues 66-75 and 81-93, and E2, consisting of residues 95-99 and 122-131) that partially overlapped with the actin-binding site of Cuc m 2. Two additional epitopes (E3, including residues 2 10, and E4, including residues 35-45) that should show weaker putative antigen antibody associations and shared most residues with synthetic peptides with low IgE-binding capacity were predicted on theoretical grounds. CONCLUSIONS: Strong and weak IgE epitopes have been uncovered in melon profilin, Cuc m 2. CLINICAL IMPLICATIONS: The different types of IgE epitopes located in the 3-dimensional structure of melon profilin can constitute the molecular basis to explain the sensitization and cross-reactivity exhibited by this panallergen family. PMID- 17397912 TI - Demographic and clinical characteristics of children and adolescents with severe or difficult-to-treat asthma. AB - BACKGROUND: Young patients with severe or difficult-to-treat asthma are an understudied population. OBJECTIVE: To assess age-associated and gender associated differences in children and adolescents in the observational study, The Epidemiology and Natural History of Asthma: Outcomes and Treatment Regimens. METHODS: Cross-sectional baseline data for patients greater than or equal to 6 years and less than or equal to 17 years (n = 1261) were stratified by age group (6-8, 9-11, 12-14, and 15-17 years). The chi(2) test for categorical variables and analysis of variance for continuous variables were used to identify differences among age groups, stratified by gender. RESULTS: Most patients had moderate (55%) or severe (41%) asthma by physician assessment. Of those using greater than or equal to 3 long-term controllers (62%), 53% of children (6-11 years) and 44% of adolescents (12-17 years) reported an oral corticosteroid burst and 25% and 19%, respectively, had an emergency department visit in the previous 3 months; 10% and 15%, respectively, reported past intubation. In females, weight for age ranged between the 67th and 70th percentiles; height for age was between the 42nd and 54th percentiles (P < .01 among age groups). Lung function was lower in adolescents than children: prebronchodilator percent predicted forced expiratory volume in 1 second (FEV(1))/forced vital capacity was 0.92 (6-8 years) and 0.83 (15-17 years), P less than .05, in males; and 0.94 (6-8 years) and 0.87 (15-17 years), P less than .05, in females. CONCLUSIONS: Children and adolescents demonstrated high rates of health care use and loss of lung function, despite using multiple long-term controllers. CLINICAL IMPLICATIONS: Asthma treatments that prevent loss of lung function and reduce health care resource use are needed in young patients with severe or difficult-to-treat asthma. PMID- 17397913 TI - Dysregulation of cardiogenesis, cardiac conduction, and cell cycle in mice lacking miRNA-1-2. AB - MicroRNAs (miRNAs) are genomically encoded small RNAs used by organisms to regulate the expression of proteins generated from messenger RNA transcripts. The in vivo requirement of specific miRNAs in mammals through targeted deletion remains unknown, and reliable prediction of mRNA targets is still problematic. Here, we show that miRNA biogenesis in the mouse heart is essential for cardiogenesis. Furthermore, targeted deletion of the muscle-specific miRNA, miR-1 2, revealed numerous functions in the heart, including regulation of cardiac morphogenesis, electrical conduction, and cell-cycle control. Analyses of miR-1 complementary sequences in mRNAs upregulated upon miR-1-2 deletion revealed an enrichment of miR-1 "seed matches" and a strong tendency for potential miR-1 binding sites to be located in physically accessible regions. These findings indicate that subtle alteration of miRNA dosage can have profound consequences in mammals and demonstrate the utility of mammalian loss-of-function models in revealing physiologic miRNA targets. PMID- 17397914 TI - SIRT1 and neuronal diseases. AB - SIRT1 is the mammalian homologue of yeast silent information regulator (Sir)-2, a member of the sirtuin family of protein deacetylases which have gained much attention as mediators of lifespan extension in several model organisms. Induction of SIRT1 expression also attenuates neuronal degeneration and death in animal models of Alzheimer's disease and Huntington's disease. SIRT1 induction, either by sirtuin activators such as resveratrol, or metabolic conditioning associated with caloric restriction (CR), could be neuroprotective in several ways. It could promote the non-amyloidogenic cleavage of the amyloid precursor protein, enhance clearance of amyloid beta-peptides, and reduced neuronal damage through potential inhibition of neuroinflammatory signaling pathways. In addition, increased SIRT1 activity could alter neuronal transcription profiles to enhance anti-stress and anti-apoptotic gene activities, and has been proposed to underlie the inhibition of axonal degeneration in the Wallerian degeneration slow (Wld(s)) phenotype. As neuronal degeneration is a major pathophysiological aspect of human aging, understanding the mechanism of SIRT1 neuroprotection promises novel strategies in clinical intervention of neurodegenerative diseases. PMID- 17397915 TI - Estimation of chemical resistance of dental ceramics by neural network. AB - OBJECTIVES: The purpose of this research was to determine the mass concentrations of ions eluted from dental ceramic after an exposure to hydrochloric acid and, drawing on those results, to develop a feedforward backpropagation neural network (NN). MATERIALS AND METHODS: Four dental ceramics were selected for this study. The experimental measurement was conducted after 1, 2, 3, 6 and 12 months of exposure to hydrochloric acid. The results of the 1, 2, 6 and 12 months of immersion were used for training a 13-13-5 model of NN. For evaluating NN efficiency, the regression analysis of input variables obtained by the experiment and output variables provided by the trained network was used. RESULTS: The measured data from the 3-month acid exposure and data obtained by the neural network estimation were compared. High correlation coefficient (R) and low normalized root mean square error (NRMSE) between the measured and estimated output values were observed. CONCLUSIONS: It could be concluded that the artificial neural network has a great potential as an additional method in investigating the properties of dental materials. PMID- 17397916 TI - On the action of ozone on gelatin. AB - Gelatin, the lower molecular weight derivative of collagen, was treated with ozone and the structural changes were studied with polarimetry, electronic absorption and FT-IR spectroscopy. The resulting ozonized gelatins were studied by thermal analysis (DSC and TGA) in comparison to a reference gelatin. It has been found that at relatively low ozone dose, for instance at ozone/gelatin molar ratios between 0.1 and 0.35 the structural damage introduced by ozone oxidation of gelatin is minimal to negligible and this may facilitate the industrial utilization of ozone in the cold sterilization of gelatin as well as in its bleaching processes required for certain applications. At higher O(3)/gelatin molar ratios the gelatine damage is evident both polarimetrically as suggested by the drop of its specific optical rotation and by spectrophotometrical analyses. PMID- 17397917 TI - Procathepsin D expression correlates with invasive and metastatic phenotype of MDA-MB-231 derived cell lines. AB - Procathepsin D (pCD) is a glycoprotein secreted abundantly by cancerous cells with a documented role in tumor development. The levels of pCD in primary tumors are highly correlated with an increased incidence of metastasis. Our earlier studies have shown that pCD exerts its effect on cancer cells through its activation peptide (AP) and involves both autocrine and paracrine modes of action. In this study, we analyzed the expression and role of pCD in MDA-MB-231 and its derived cell lines 1833 and 4175 possessing discrete metastatic abilities. Our results demonstrated a direct relationship between expression and secretion of pCD to the differential invasive potential of these cells. Also, the cell lines responded to AP treatment by enhancing their invasive potential, proliferation and induction of secretion of various cytokines, suggesting that pCD plays a role in metastasis through its AP region. PMID- 17397918 TI - Photo-patterning of porous hydrogels for tissue engineering. AB - Since pore size and geometry strongly impact cell behavior and in vivo reaction, the ability to create scaffolds with a wide range of pore geometries that can be tailored to suit a particular cell type addresses a key need in tissue engineering. In this contribution, we describe a novel and simple technique to design porous, degradable poly(2-hydroxyethyl methacrylate) hydrogel scaffolds with well-defined architectures using a unique photolithography process and optimized polymer chemistry. A sphere-template was used to produce a highly uniform, monodisperse porous structure. To create a patterned and porous hydrogel scaffold, a photomask and initiating light were employed. Open, vertical channels ranging in size from 360+/-25 to 730+/-70 microm were patterned into approximately 700 microm thick hydrogels with pore diameters of 62+/-8 or 147+/ 15 microm. Collagen type I was immobilized onto the scaffolds to facilitate cell adhesion. To assess the potential of these novel scaffolds for tissue engineering, a skeletal myoblast cell line (C2C12) was seeded onto scaffolds with 147 microm pores and 730 microm diameter channels, and analyzed by histology and digital volumetric imaging. Cell elongation, cell spreading and fibrillar formation were observed on these novel scaffolds. In summary, 3D architectures can be patterned into porous hydrogels in one step to create a wide range of tissue engineering scaffolds that may be tailored for specific applications. PMID- 17397919 TI - The effect of surface charge on the uptake and biological function of mesoporous silica nanoparticles in 3T3-L1 cells and human mesenchymal stem cells. AB - Cellular uptake of nanoparticles for stem cell labeling/tracking is considered as the most promising method. Recently mesoporous silica nanoparticles (MSNs) are emerging as an idea agent for efficient stem cell labeling. The objective of this study was to evaluate the effect of surface charge on the highly efficient cellular uptake and in vitro cytotoxicity of MSNs in human mesenchymal stem cells (hMSCs). The surface charge was varied by the degree of surface modification with N-trimethoxysilylpropyl-N,N,N-trimethylammonium chloride and the uptake of MSNs was detected by flow cytometry. 3T3-L1 cells were also used to compare the uptake behavior of MSNs between cell types. A clear correlation of positive surface charge and the number of fluorescence-labeled cells was mainly observed in 3T3-L1 cells. In both cells, uptake of unmodified MSNs was inhibited by phenylarsine oxide (PAO) and cytochalasin D (Cyt D) suggesting a clathrin- and an actin dependent endocytosis were involved. With strongly positive-charged MSNs, the inhibitory effects were observed in 3T3-L1 cells but not in hMSCs. Without regard to the surface charge, uptake of MSNs into both cells did not affect their viability, proliferation, and differentiation. Our results show that MSNs uptake by hMSCs can be regulated by a threshold of positive surface charge but also imply that the modulation of surface charge on MSNs uptake is specific to cell type. PMID- 17397920 TI - Reduction of immunoglobulin levels during imatinib therapy of chronic myeloid leukemia. PMID- 17397921 TI - Monocytosis is an adverse prognostic factor for survival in younger patients with primary myelofibrosis. AB - We recently developed a modified Dupriez prognostic scoring system (PSS) that effectively discriminated between high-, intermediate-, and low-risk young patients (age < or =60 years) with primary myelofibrosis (PMF) based on the respective presence of none, one, or two or more of the following parameters: hemoglobin <10 g/dL, leukocyte count <4 or >30 x 10(9)L(-1), and platelet count <100 x 10(9)L(-1). The current study (n=129; median age, 52 years; 69 males) reveals, on multivariable analysis, that an absolute monocyte count of > or =1 x 10(9)L(-1) carries an independent predictive value (p=0.02), for an inferior survival, in addition to that provided by hemoglobin level (p=0.002), platelet count (0.02), and leukocyte count (p=0.16). The inclusion of the monocyte count as a fourth risk factor enabled the construction of a new and improved Mayo PSS; median survival was 173, 61, and 26 months in the absence of all four (low-risk), three (intermediate-risk), or two or less (high-risk) adverse features, respectively (p<0.0001). The independent prognostic value of monocytosis was validated in a separate database of 97 patients with PMF from another institution. PMID- 17397922 TI - Involvement of c-Jun N-terminal kinase in G2/M arrest and FasL-mediated apoptosis induced by a novel indoloquinoline derivative, IQDMA, in K562 cells. AB - N'-(11H-Indolo[3,2-c]quinolin-6-yl)-N,N-dimethylethane-1,2-diamine (IQDMA), an indoloquinoline derivative, synthesized in our laboratory, has been demonstrated to be an effective anti-tumor agent in human leukemia cells. Treatment of K562 cells with IQDMA resulted in G2/M phase cell cycle arrest, presumably involving the concomitant up-regulation of p21 and apoptosis through up-regulation of FasL and sequential activation of caspase-8 and caspase-3. In contrast to the lack of appreciable effect on the phosphorylation of ERK and p38 MAPK, activation of JNK was noted when K562 cells were exposed to IQDMA. Moreover, IQDMA-mediated G2/M phase arrest and apoptosis were reversed after treatment with the JNK-specific inhibitors, SP600125 and JNK inhibitor 1. Further investigation showed that SP600125 reduced the activation of FasL, caspase-3, caspase-8, and led to a marked decline of p21. Taken together, our data show that JNK plays an important role in IQDMA-mediated G2/M arrest and apoptosis of K562 cancer cells. PMID- 17397923 TI - Interventions for branch retinal vein occlusion: an evidence-based systematic review. AB - TOPIC: To assess the evidence on interventions to improve visual acuity (VA) and to treat macular edema and/or neovascularization secondary to branch retinal vein occlusion (BRVO). CLINICAL RELEVANCE: Branch retinal vein occlusion is the second most common retinal vascular disease. METHODS/LITERATURE REVIEWED: English and non-English articles were retrieved using a keyword search of Medline (1966 onwards), Embase, the Cochrane Collaboration, the National Institute of Health Clinical Trials Database, and the Association for Research in Vision and Ophthalmology Annual Meeting Abstract Database (2003-2005). This was supplemented by hand searching references of review articles. Two investigators independently identified all randomized clinical trials (RCTs) with more than 3 months' follow up. RESULTS: From 4332 citations retrieved, 12 RCTs were identified. There were 5 RCTs on laser photocoagulation. Grid macular laser photocoagulation was effective in improving VA in 1 large multicenter RCT, the Branch Vein Occlusion Study (BVOS), but 2 smaller RCTs found no significant difference. The BVOS showed that scatter retinal laser photocoagulation was effective in preventing neovascularization and vitreous hemorrhage in patients with neovascularization, but a subsequent RCT found no significant effect. Randomized clinical trials evaluating intravitreal steroids (n = 2), hemodilution (n = 3), ticlopidine (n = 1), and troxerutin (n = 1) showed limited or no benefit. CONCLUSIONS: There is limited level I evidence for any interventions for BRVO. The BVOS showed that macular grid laser photocoagulation is an effective treatment for macular edema and improves vision in eyes with VA of 20/40 to 20/200, and that scatter laser photocoagulation can effectively treat neovascularization. The effectiveness of many new treatments is unsupported by current evidence. PMID- 17397924 TI - Randomized comparison of the intraocular pressure-lowering effect of phacoviscocanalostomy and phacotrabeculectomy. AB - PURPOSE: To compare the intraocular pressure (IOP)-lowering effect of combined viscocanalostomy and phacoemulsification and combined trabeculectomy and phacoemulsification with mitomycin C in eyes with primary open-angle glaucoma. DESIGN: Prospective randomized 1-year trial. PARTICIPANTS: Forty consecutive patients (40 eyes) with primary open-angle glaucoma and cataract. INTERVENTION: Eyes were assigned randomly either to trabeculectomy with mitomycin C or to viscocanalostomy in combination with phacoemulsification and intraocular lens implantation. MAIN OUTCOME MEASURES: Success rate based on IOP. RESULTS: Mean baseline IOP was 24.0+/-2.0 mmHg in the viscocanalostomy group and 23.7+/-2.6 mmHg in the trabeculectomy group (P = 0.7). Mean postoperative IOP was 13.7+/-2.2 mmHg at 3 months, 14.8+/-3.3 mmHg at 6 months, and 14.9+/-3.0 mmHg at 12 months in the viscocanalostomy group and 12.1+/-4.0 mmHg at 3 months, 13.8+/-4.7 mmHg at 6 months, and 14.1+/-4.4 mmHg at 12 months in the trabeculectomy group. There was no significant difference in the mean IOP between the groups at any time. At 12 months, 17 patients (85%) in the viscocanalostomy group and 16 patients (80%) in the trabeculectomy group achieved an IOP of 20 mmHg or less without medication (P = 0.7). Complications included 2 cases (10%) of flat/shallow anterior chamber and 4 cases (20%) of hypotony in the trabeculectomy group, whereas intraoperative microperforation of Descemet's membrane occurred in 3 cases (15%) in the viscocanalostomy group. CONCLUSIONS: There was no significant difference in IOP reduction between viscocanalostomy and trabeculectomy with mitomycin C in combination with phacoemulsification and intraocular lens implantation in patients with primary open-angle glaucoma. PMID- 17397925 TI - Nine-year incidence of open-angle glaucoma in the Barbados Eye Studies. AB - PURPOSE: To determine the 9-year incidence of open-angle glaucoma (OAG) in African-descent participants of the Barbados Eye Studies. DESIGN: Nine-year cohort study with 81% to 85% participation. PARTICIPANTS: Three thousand two hundred twenty-two persons without definite OAG at baseline, at risk of developing OAG at follow-up. METHODS: The standardized protocol included automated perimetry and various ophthalmic measurements, with a comprehensive ophthalmologic examination for those referred. Fundus photographs were evaluated independently by masked graders. Incidence was estimated by the product-limit approach. Relative risk (RR) ratios with 95% confidence intervals (CIs) were based on Cox regression models with discrete time. MAIN OUTCOME MEASURE: Nine year incidence of definite OAG, based on the development of visual field defects and glaucomatous optic neuropathy, with ophthalmologic confirmation. RESULTS: The 9-year incidence of definite OAG was 4.4% (95% CI, 3.7%-5.2%), or an average of 0.5%/year, based on 125 new cases. Incidence increased greatly with age, from 2.2% at ages 40 to 49 years to 7.9% at ages 70 years or older, and tended to be higher in men than women (4.9% vs. 4.1%; RR, 1.3; 95% CI, 0.9-1.8). More than half (53%) of new cases were undetected, and of these, one third had intraocular pressure of 21 mmHg or less. When 141 persons developing suspected/probable OAG were considered, the total incidence was 9.4% (8.4%-10.6%), averaging approximately 1%/year, also increasing with age, and significantly higher in men than women (10.7% vs. 8.6%; RR, 1.31; 95% CI, 1.02-1.67). CONCLUSIONS: These new data provide a measure of the long-term risk of OAG in an African-descent population, which is markedly higher than in persons of European ancestry. Results confirm the increased risk with age and in men. The incidence data fill a gap in our understanding of OAG risk and have implications for public health policy and planning; they also will allow the study of factors related to the risk of OAG development. PMID- 17397926 TI - Frequency doubling perimetry and short-wavelength automated perimetry to detect early glaucoma. AB - PURPOSE: To compare the ability of short-wavelength automated perimetry (SWAP) and frequency doubling perimetry (FDP) to detect early glaucoma damage. DESIGN: Prospective case-control study using a comparative case series. PARTICIPANTS: Forty-two patients with preperimetric glaucomatous optic nerve damage and a normal standard achromatic perimetry (SAP) in 1 eye, but with contralateral SAP abnormalities, and 35 normals. METHODS: Forty-two patients and 35 normals underwent SWAP and FDP (Humphrey 24-2; Carl Zeiss Meditec, Inc., Dublin, CA). Correlations of mean deviation (MD) and pattern standard deviation (PSD) of the 2 groups were calculated. The number of defects at P<0.05 and P<0.01 on total deviation (TD) and pattern deviation (PD) plots were compared. Diagnostic precision and agreement on location of abnormalities were determined. MAIN OUTCOME MEASURES: Correlations and comparisons of global indices between SWAP and FDP: MD, PSD, TD, and PD abnormal number of points. RESULTS: Significant correlations in the glaucoma group were found between SWAP and FDP for MD (r = 0.54; P<0.008) and PSD (r = 0.49; P<0.001). Defects on the TD and PD plots were detected more frequently by FDP in the glaucoma group, although they were significant only for PD at P<0.01 (P = 0.024). Areas under receiver operator characteristic curves for MD of SWAP and PSD of FDP were 0.74 and 0.67, respectively (P = 0.37). Using defined defect criteria, FDP had a significantly higher sensitivity (72% vs. 54%; P = 0.02) and similar specificity (53% vs. 44%; P = 0.12) compared with SWAP. Agreement on defect location was moderate (kappa, 0.46). Testing time for SWAP was longer than for FDP in both glaucomatous and normal eyes (P<0.001). CONCLUSIONS: Short-wavelength automated perimetry and FDP showed similar ability to detect visual dysfunction in patients with preperimetric glaucoma. Long-term follow-up is required to define their role in predicting subsequent SAP defects. PMID- 17397927 TI - Modulation of behavior and NMDA-R1 gene mRNA expression in adult female mice after sub-acute administration of benzo(a)pyrene. AB - The behavioral performances of adult mice exposed to sub-acute doses of benzo(a)pyrene (B(a)P) were monitored in tests related to learning and memory (Y maze and Morris water maze), locomotor activity (open-field test) and motor coordination (Locotronic apparatus). At low doses (0.02 and 0.2mg/kg), B(a)P impaired short-term learning and spatial memory performance in the Y maze and in the Morris water maze tests. Surprisingly, in the Y maze, the performances of animals exposed to the highest dose of B(a)P (200mg/kg) were quite similar to those of control animals. Hyperactivity/hyperarousal observed in both tests at this dose and attributed to an anxiolytic-like effect of B(a)P may have blurred the learning deficit in these mice faced with a new situation. These deficits seem to be unrelated to motor impairments because B(a)P had no effect on locomotor activity and motor coordination. We demonstrated that sub-acute exposure to B(a)P in adult mice also modulates gene expression of NMDA-R1 subunit in brain areas highly involved in cognitive function like the hippocampus, suggesting a relationship between the expression of functional NMDA-R1 mRNA, impairment of short-term and spatial memory and the B(a)P exposure levels. PMID- 17397928 TI - Another fundamental error in "What is the meaning of non-linear dose-response relationships between blood lead concentrations and IQ?" became obvious in the authors' response to comments. PMID- 17397929 TI - Synthesis, X-ray crystal structures and biomimetic and anticancer activities of novel copper(II)benzoate complexes incorporating 2-(4'-thiazolyl)benzimidazole (thiabendazole), 2-(2-pyridyl)benzimidazole and 1,10-phenanthroline as chelating nitrogen donor ligands. AB - Cu(BZA)(2)(EtOH)(0.5) (1) was generated by the reaction of copper(II) hydroxide with benzoic acid (BZAH). [Cu(TBZH)(2)(BZA)](BZA).0.5TBZH.H(2)O (2) and [Cu(2 PyBZIMH)(2-PyBZIM)(BZA)].1.66EtOH (3) were obtained when 1 reacted with Thiabendazole (TBZH) and 2-(2-pyridyl)benzimidazole (2-PyBZIMH), respectively. [Cu(BZA)(2)(phen)(H(2)O)] (4) was isolated from the reaction of benzoic acid and 1,10-phenanthroline (phen) with copper(II)acetate dihydrate. Molecular structures of 2, 3 and 4 were determined crystallographically. 2 and 3 are hydrogen bonded dimers and trimers, respectively. The copper centres in complexes 2 and 3 are bis chelate derivatives that have N(4)O ligation and their geometry is very similar being approximately square-pyramidal. However whereas in complex 2 both TBZH ligands are neutral in 3 one of the 2-PyBZIMH chelators is deprotonated on each copper. The structural results for 4 represent a re-examination of this crystallographically known compound for which no hydrogen atom coordinates have been previously reported. It crystallises as a hydrogen bonded dimmer and is a mono-chelate of phen with each copper centre possessing N(2)O(3) ligation and square pyramidal geometry. The catalase and superoxide dismutase (SOD) activities of the four complexes along with those of the known phenanthroline complexes [Cu(mal)(phen)(2)] and [Cu(phendione)(3)](ClO(4))(2) (malH(2)=malonic acid and phendione=1,10-phenanthroline-5,6-dione) were investigated. Complexes 1-4, the metal free ligands and a simple copper(II) salt were assessed for their cancer chemotherapeutic potential against the hepatocellular carcinoma (Hep-G(2)) and kidney adenocarcinoma (A-498) cell lines. TBZH, 2-PyBZIMH and benzoic acid when uncoordinated to a metal centre offer poor chemotherapeutic potential. copper(II) benzoate is significantly more active than the free acid. The bis-chelate derivatives [Cu(TBZH)(2)(BZA)](BZA).0.5TBZH.H(2)O (2) and [Cu(2-PyBZIMH)(2 PyBZIM)(BZA)].1.66EtOH (3) elicit a significant cytotoxic response to the cancer cell lines tested. Replacing TBZH and 2-PyBZIMH with phen to give [Cu(BZA)(2)(phen)(H(2)O)] (4) does not significantly increase the anti-cancer activity. PMID- 17397931 TI - Lentiviral vector-mediated genetic modification of human neural progenitor cells for ex vivo gene therapy. AB - Human neural progenitor cells (hNPC) hold great potential as an ex vivo system for delivery of therapeutic proteins to the central nervous system. When cultured as aggregates, termed neurospheres, hNPC are capable of significant in vitro expansion. In the current study, we present a robust method for lentiviral vector mediated gene delivery into hNPC that maintains the differentiation and proliferative properties of neurosphere cultures while minimizing the amount of viral vector used and controlling the number of insertion sites per population. This method results in long-term, stable expression even after differentiation of the hNPC to neurons and astrocytes and allows for generation of equivalent transgenic populations of hNPC. In addition, the in vitro analysis presented predicts the behavior of transgenic lines in vivo when transplanted into a rodent model of Parkinson's disease. The methods presented provide a powerful tool for assessing the impact of factors such as promoter systems or different transgenes on the therapeutic utility of these cells. PMID- 17397932 TI - Long-term functional and morphological assessment of a standardized rat sciatic nerve crush injury with a non-serrated clamp. AB - We have recently described the sequence of functional and morphologic changes occurring after a standardized sciatic nerve crush injury. An 8-week post-injury time was used because this end point is the far most used. Unexpectedly, both functional and morphological data revealed that animals had still not recovered to normal pre-injury levels. Therefore, the present study was designed in order to prolong the observation up to 12 weeks. Functional recovery was evaluated using sciatic functional index (SFI), static sciatic index (SSI), extensor postural thrust (EPT), withdrawal reflex latency (WRL) and ankle kinematics. In addition, quantitative morphology was carried out on regenerated nerve fibers. A full functional recovery was predicted by SFI/SSI, EPT and WRL but not all ankle kinematics parameters. Moreover, only two morphological parameters (myelin thickness/axon diameter ratio and fiber/axon diameter ratio) returned to normal values. Data presented in this paper provide a baseline for selecting the adequate end-point and methods of recovery assessment for a rat sciatic nerve crush study and suggest that the combined use of functional and morphological analysis should be recommended in this experimental model. PMID- 17397930 TI - Astrocyte elevated gene-1: recent insights into a novel gene involved in tumor progression, metastasis and neurodegeneration. AB - Tumor progression and metastasis are complex processes involving intricate interplay among multiple gene products. Astrocyte elevated gene (AEG)-1 was cloned as an human immunodeficiency virus (HIV)-1-inducible and tumor necrosis factor-alpha (TNF-alpha)-inducible transcript in primary human fetal astrocytes (PHFA) by a rapid subtraction hybridization approach. AEG-1 down-regulates the expression of the glutamate transporter EAAT2; thus, it is implicated in glutamate-induced excitotoxic damage to neurons as evident in HIV-associated neurodegeneration. Interestingly, AEG-1 expression is elevated in subsets of breast cancer, glioblastoma multiforme and melanoma cells, and AEG-1 cooperates with Ha-ras to augment the transformed phenotype of normal immortal cells. Moreover, AEG-1 is overexpressed in >95% of human malignant glioma samples when compared with normal human brain. Overexpression of AEG-1 increases and siRNA inhibition of AEG-1 decreases migration and invasion of human glioma cells, respectively. AEG-1 contains a lung-homing domain facilitating breast tumor metastasis to lungs. These findings indicate that AEG-1 might play a pivotal role in the pathogenesis, progression and metastasis of diverse cancers. Our recent observations indicate that AEG-1 exerts its effects by activating the nuclear factor kappa B (NF-kappaB) pathway and AEG-1 is a downstream target of Ha-ras and plays an important role in Ha-ras-mediated tumorigenesis. These provocative findings are intensifying interest in AEG-1 as a crucial regulator of tumor progression and metastasis and as a potential mediator of neurodegeneration. In this review, we discuss the cloning, structure and function(s) of AEG-1 and provide recent insights into the diverse actions and intriguing properties of this molecule. PMID- 17397933 TI - Artifactual dendritic beading in rat spinal cord induced by perfusion with cold saline and paraformaldehyde. AB - Extensive dendritic beading of MAP2 (microtubule-associated protein 2) immunoreactivity has previously been observed in the contused rat spinal cord. However, we have also observed dendritic beading in occasional uninjured animals. The purpose of this study was to examine the possibility that perfusion conditions contributed to the dendritic beading. Under deep anesthesia, uninjured rats (adult female Long-Evans, 200-225 g) were transcardially perfused with 0.9% saline solution followed by 4% paraformaldehyde at cold (4 degrees C) or warm (20 degrees C) temperature, and at a low (20 ml/min) or high (50 ml/min) flow rate. Dendrites were visualized by MAP2 immunoreactivity. The results demonstrate that perfusion with cold solutions at a high flow rate induces pronounced dendritic beading, and when perfused at a low flow rate, results in moderate dendritic beading. Warm perfusates did not induce dendritic beading when administered at a low flow rate, but occasional beading was observed with a high flow rate. Western blots revealed spectrin breakdown, but not MAP2 loss, in rats perfused with cold saline solution at a high flow rate, conditions that also resulted in dendritic beading. These findings demonstrate that dendritic morphology is sensitive to both temperature and flow rate of the perfusate. Warm fixative and a low perfusion flow rate minimized the perfusion-induced dendritic beading. PMID- 17397934 TI - Behind impulsive suicide attempts: indications from a community study. AB - AIM: A considerable proportion of suicide attempts are made on impulse. However, knowledge of characteristics of impulsive attempters is still limited. The present study investigated some of these characteristics and aimed to identify the pattern (if any) of suicidal ideation before an impulsive attempt. METHODS: Data from a randomized and stratified population of 5130 individuals from Brisbane, Australia, were analysed. Computer-assisted telephone interviews (CATI) were adopted to recruit subjects. Those reporting previous suicidal behaviour were sent a questionnaire by mail. RESULTS: One hundred and twelve subjects reported a suicide attempt. One quarter of these described a pattern consistent with an impulsive attempt. Most impulsive attempters experienced suicidal thoughts before their attempt. They were less likely to believe that their attempt would cause death, and less likely to experience depression. Impulsive attempters did not differ significantly from non-impulsive attempters in regards to age, gender, and motivations for the attempt. Surprisingly, no differences in mean scores of trait impulsivity between impulsive and non-impulsive attempters were found. In addition, the majority of suicide attempters (whether impulsive or not) experienced the suicidal process as fluctuating and not as developing along a continuum. LIMITATIONS: The number of attempters who validly entered the study limited our ability to identify potential confounders. Due to the retrospective nature of the survey, the reliability of the information collected may have been affected by recall biases. In addition, as the surveys were administered by mail, it is possible that some questions may have been misinterpreted. CONCLUSIONS: The presence of suicidal feelings prior to an attempt constitutes an opportunity for intervention also in impulsive attempters. However, the identification of impulsiveness requires more research efforts. PMID- 17397935 TI - Interactions between Streptococcus suis serotype 2 and cells of the myeloid lineage in the palatine tonsil of the pig. AB - Streptococcus suis is a major pathogen in pigs and causes significant morbidity and mortality in herds world-wide. A major problem with S. suis is the presence of asymptomatic carrier animals which can spread the organism within and between herds. The palatine tonsil is one of the main sites where the organism can be recovered, both in infected and carrier animals. The use of multiple-colour immunohistology allowed identification of the cell types associated with bacteria in the tonsils of infected gnotobiotic piglets. Bacteria were never associated with T-cells or B-cells but were always associated with cells of the myeloid lineage. Expression of CD16 and CD163 on these leukocytes suggested an association with mature macrophages in tonsil, which may lead to clearance or control of the micro-organism. PMID- 17397936 TI - Apoptosis: a target for potentiation of UV-induced IL-1Ra synthesis by IVIg. AB - IL-1Ra prevents IL-1 induced inflammatory signalling, a mechanism potentially important for several pathological conditions characterized by inflammation. When administered as a drug in the recombinant form, it displays a protective effect towards them. We postulated that this action could also be achieved by pharmacological activation of endogenous IL-1Ra production. We previously showed that photochemotherapy and UV-light increased monocyte/macrophages IL-1Ra secretion. A similar effect has been shown for IVIg. The aim of this study was to define optimal in vitro conditions for induction of IL-1Ra secretion. As both agents induce lymphocytes apoptosis, we focused our analysis on the influence of IVIg on UV-induced-IL-1Ra production on this mechanism. After overnight preincubation at 37 degrees C, UV-irradiated PBL mixed with two IVIg concentrations (1 and 25 mg/ml) were cocultured with autologous PBMC. Apoptosis was measured by annexin V/PI. IL-1Ra secretion was evaluated by RT-PCR and Luminex microbead array assay. A significant additive dose-dependent influence of IVIg (+85%; p=0.0005) on UV-induced IL-1Ra secretion, involved both Fc-receptor activation at a low dose (1 mg/ml) and a potent apoptotic action on PBL reinforcing the UV effect at high concentrations (25 mg/ml). We conclude that lymphocyte apoptosis represents an important pathway contributing to enhancement of UV-induced monocyte/macrophages IL-1Ra production by IVIg and that these findings should be considered when evaluating in vivo protocols for photochemotherapy and IVIg treatment, in hope of improving efficacy. PMID- 17397937 TI - Implementation of a one-step real-time RT-PCR protocol for diagnosis of foot-and mouth disease. AB - An automated one-step real-time reverse transcription polymerase chain reaction (rRT-PCR) protocol was optimised and evaluated for the routine diagnosis of foot and-mouth disease (FMD). Parallel testing of RNA samples (n=257) indicated that this assay has a diagnostic sensitivity at least equivalent to the automated two step rRT-PCR protocol previously used for the laboratory detection of FMD virus (FMDV). This more rapid and economical one-step protocol will play a key role in contingency planning for any future outbreaks of FMD in the United Kingdom (UK). PMID- 17397938 TI - Fifty years of dopamine research. PMID- 17397939 TI - In vitro resistance to interferon-alpha of hepatitis B virus with basic core promoter double mutation. AB - The hepatitis B virus (HBV) genome basic core promoter (BCP) modulates HBeAg secretion at the transcriptional level. In addition to pre-core mutations, variations in the BCP are related to hepatitis B e antigen (HBeAg)-negative chronic hepatitis B. HBeAg-negative chronic hepatitis B patients show a lower sustained response to interferon (IFN). The aim of this study was to determine if there is a relationship between HBV BCP mutation and sensitivity of HBV to IFN alpha in vitro. BCP mutations were introduced by site-directed mutagenesis and the entire genomes of wild-type and mutant HBV were transiently transferred into Huh7 cells by calcium phosphate transfection. With or without IFN-alpha, viral products in the culture medium and viral replication intermediates in the cytoplasm were detected 3 days after transfection. The amount of hepatitis B surface antigen (HBsAg) secreted by wild-type HBV and the BCP mutant was similar, while HBeAg secreted by the mutant was decreased by 35.4%. HBV particles and replication intermediates of the BCP mutant were increased. After IFN-alpha was added, HBeAg, HBV DNA and HBV replication intermediates decreased for both the wild-type HBV (by 25.7%, 31.8%, 29.8%, respectively) and the BCP mutant (by 8.4%, 27.4%, 10.1%, respectively). These data indicate that HBV harboring the BCP double mutation has stronger replication competence and lower sensitivity to IFN alpha than wild-type. PMID- 17397940 TI - Qualitative changes in ultrasonic vocalization in rats after unilateral dopamine depletion or haloperidol: a preliminary study. AB - The sensorimotor speech/voice deficits associated with Parkinson disease have been well documented in humans. They are largely resistant to pharmacological and surgical treatment, but respond to intensive speech therapy. The mechanisms underlying this phenomenon are not well understood and are difficult to systematically test in humans. Thus, we turn to the rat as a model. The purpose of this study is to compare the ultrasonic vocalization (USV) of rats in three conditions: control, haloperidol-induced transient dopamine depletion, and unilateral 6-hydroxydopamine (6-OHDA) induced moderately-severe degeneration of dopamine neurons. It was hypothesized that both dopamine-altered conditions would lead to a change in the features of the USV acoustic signal. Results demonstrated that bandwidth decreased in the dopamine-altered rats. This is the first study to document a degradation of the acoustic signal of frequency-modulated 50-kHz calls as a result of interfering with dopamine synaptic transmission in rats. The data suggest that mild transient dopamine depletion with haloperidol or even unilateral degeneration of dopamine neurons is associated with changes in the USV acoustic signal. Dopaminergic dysfunction influences USV quality without reducing the number of calls. This study provides a foundation to examine the role of dopamine in sensorimotor processes underlying USV production and potentially to explore treatments for dopamine deficiency-related impaired vocal outcome. PMID- 17397941 TI - Enriched environments influence depression-related behavior in adult mice and the survival of newborn cells in their hippocampi. AB - Major depression is a highly prevalent mental disorder and environmental factors have been strongly implicated in its pathophysiology. Clinical studies have demonstrated that stress or depression can lead to atrophy and cell loss in the hippocampus. Studies of animal models of depression have suggested that reduced neurogenesis in the adult hippocampus might contribute to such structural changes and to the behavior of these animals. On the other hand, increased hippocampal neurogenesis can be induced by the administration of antidepressants or electroconvulsive seizure, suggesting that increased neurogenesis might be related to the treatment of depression. Thus, an enriched environment (EE), which also enhances neurogenesis, is expected to have therapeutic effects on depression related behaviors. To investigate the effects of an EE during adulthood on these behaviors, we subjected adult mice housed in an EE for five weeks to behavioral tests. In an open field test, EE mice exhibited a decrease in the distance traveled and an increase in the amount of time spent in the center. The startle response was smaller in EE mice than in control mice. EE mice also showed reduced immobility time in a forced swim test. The immobility time in EE mice was approximately half that observed in mice treated with a tricyclic antidepressant, imipramine. In our experimental condition, increased survival of newborn cells was observed in EE mice by 5-bromo-2'-deoxyuridine (BrdU)-labeled immunohistochemistry. Double-staining of BrdU and a mature neuron marker, NeuN, revealed that the majority of surviving cells were neurons. Our results suggest that EE, which enhanced the survival of newborn neurons, shows beneficial effects on behavioral despair and habituation to a novel environment. PMID- 17397942 TI - Mu opioid receptor knockout mice in the Morris Water Maze: a learning or motivation deficit? AB - An earlier study done by Kas et al. [Kas MJ, van den Bos R, Baars AM, Lubbers M, Lesscher HM, Hillebrand JJ, et al. Mu-opioid receptor knockout mice show diminished food-anticipatory activity. Eur J Neurosci 2004;20(6):1624-32] suggested that mu opioid receptor (MOR) knockout mice have deficits in the motivational component rather than in the information processing component of learning. To substantiate this difference further, MOR knockout mice and wildtype littermates were tested in the Morris Water Maze (MWM), which allows for testing both components of learning. On traditional parameters for performance, no significant differences between genotypes were found. However, swimming velocity, indicative of motivation, decreased for MOR knockout mice during the course of training but not for wildtype mice. In contrast, probe trial performance was comparable between genotypes. Again, these results suggest normal information processing abilities but a decreased motivation in this MOR knockout mouse. Our conclusion is discussed in the light of other studies using MOR knockout mice that do find differences on information processing between genotypes in MWM performance. PMID- 17397943 TI - Hyperactivity in the rat is associated with spontaneous low level of n-3 polyunsaturated fatty acids in the frontal cortex. AB - Inattention, hyperactivity and impulsiveness are the main symptoms of the heterogeneous attention-deficit/hyperactivity disorder (ADHD). It has been suggested that ADHD is associated with an imbalance in polyunsaturated fatty acid (PUFA) composition, with abnormal low levels of the main n-3 PUFA, DHA (22: 6n 3). DHA is highly accumulated in nervous tissue membranes and is implicated in neural function. Animal studies have shown that diet-induced lack of DHA in the brain leads to alterations in cognitive processes, but the relationship between DHA and hyperactivity is unclear. We examined the membrane phospholipid fatty acid profile in frontal cortex of rats characterized for attention, impulsiveness and motricity in various environmental contexts to determine the relationship between brain PUFA composition and the symptoms of ADHD. The amounts of n-3 PUFA in the PE were significantly correlated with nocturnal locomotor activity and the locomotor response to novelty: hyperactive individuals had less n-3 PUFA than hypoactive ones. We conclude that spontaneous hyperactivity in rats is the symptom of ADHD that best predicts the n-3 PUFA content of the frontal cortex. This differential model in rats should help to better understand the role of PUFA in several psychopathologies in which PUFA composition is modified. PMID- 17397944 TI - C-terminal proteolysis of prenylated proteins in trypanosomatids and RNA interference of enzymes required for the post-translational processing pathway of farnesylated proteins. AB - The C-terminal "CaaX"-motif-containing proteins usually undergo three sequential post-translational processing steps: (1) attachment of a prenyl group to the cysteine residue; (2) proteolytic removal of the last three amino acids "aaX"; (3) methyl esterification of the exposed alpha-carboxyl group of the prenyl cysteine residue. The Trypanosoma brucei and Leishmania major Ras converting enzyme 1 (RCE1) orthologs of 302 and 285 amino acids-proteins, respectively, have only 13-20% sequence identity to those from other species but contain the critical residues for the activity found in other orthologs. The Trypanosoma brucei a-factor converting enzyme 1 (AFC1) ortholog consists of 427 amino acids with 29-33% sequence identity to those of other species and contains the consensus HExxH zinc-binding motif. The trypanosomatid RCE1 and AFC1 orthologs contain predicted transmembrane regions like other species. Membranes from Sf9 cells expressing the RCE1 ortholog of T. brucei or L. major showed proteolytic activity against farnesylated RAS-CVIM, whereas membranes containing T. brucei AFC1 ortholog were inactive. The results suggest that RCE1 is responsible for proteolytic removal of the C-terminal aaX from prenyl-CaaX proteins in these parasites. All the three enzymatic post-translational processes are thought to be required for proper cellular functioning of CaaX-proteins in eukaryotic cells. We carried out RNA interference experiments in Trypanosoma brucei of the enzymes involved in farnesyl protein post-translational modification to evaluate their importance in cell proliferation. Knockdown of T. brucei PFT beta subunit and RCE1 mRNAs resulted in >20-fold suppression of cell growth and dramatic morphologic changes. Knockdown of PPMT mRNA caused less dramatic effects on growth but induced noticeable changes in cell morphology. PMID- 17397945 TI - The direct p53 target gene, FLJ11259/DRAM, is a member of a novel family of transmembrane proteins. AB - The tumor suppressor p53 regulates diverse biological processes primarily via activation of downstream target genes. Even though many p53 target genes have been described, the precise mechanisms of p53 biological actions are uncertain. In previous work we identified by microarray analysis a candidate p53 target gene, FLJ11259/DRAM. In this report we have identified three uncharacterized human proteins with sequence homology to FLJ11259, suggesting that FLJ11259 is a member of a novel family of proteins with six transmembrane domains. Several lines of investigation confirm FLJ11259 is a direct p53 target gene. p53 siRNA prevented cisplatin-mediated up-regulation of FLJ11259 in NT2/D1 cells. Likewise in HCT116 p53+/+ cells and MCF10A cells, FLJ11259 is induced by cisplatin treatment but to a much lesser extent in isogenic p53-suppressed cells. A functional p53 response element was identified 22.3 kb upstream of the first coding exon of FLJ11259 and is shown to be active in reporter assays. In addition, chromatin immunoprecipitation assays indicate that p53 binds directly to this element in vivo and that binding is enhanced following cisplatin treatment. Confocal microscopy showed that an FLJ-GFP fusion protein localizes mainly in a punctate pattern in the cytoplasm. Overexpression studies in Cos-7, Saos2, and NT2/D1 cells suggest that FLJ11259 is associated with increased clonal survival. In summary, we have identified FLJ11259/DRAM as a p53-inducible member of a novel family of transmembrane proteins. FLJ11259/DRAM may be an important modulator of p53 responses in diverse tumor types. PMID- 17397946 TI - Isolated primary cardiac amyloidosis associated with porokeratosis of Mibelli. AB - In this report we briefly describe a 54-year-old woman who was referred to our institution for evaluation and management of newly diagnosed congestive heart failure associated with a skin rash. Detailed investigations revealed the presence of restrictive cardiomyopathy due to isolated primary cardiac amyloidosis as well as the presence of a skin disease named 'porokeratosis of Mibellli'. Interestingly, porokeratotic lesions rarely have been associated with localized cutaneous amyloidosis. Presumably, porokeratosis induces secondary dermal amyloid deposition by a yet unknown mechanism. This is the first case of primary amyloidosis associated with porokeratosis reported in the literature. PMID- 17397947 TI - Progression of dissection due to residual dissection after intracoronary stenting for spontaneous coronary dissection at bifurcation site of LAD and diagonal artery. AB - A 34-year old male patient visited our hospital due to severe chest pain. Initial ECG showed ST elevation at precordial leads and all cardiac enzymes were markedly elevated. Coronary angiography showed a long, longitudinal coronary dissection with dissected flap extending from the proximal LAD to the mid segment of the vessel and proximal diagonal artery. IVUS showed dissected flap and false lumen communicating with true lumen from proximal to mid-LAD. We implanted two paclitaxel-eluting stents using crushing technique at bifurcation lesion and overlapped another paclitaxel-eluting stent at proximal LAD for full coverage of dissection. Final angiography showed good distal flow. However, despite of maximal pressure of post stent ballooning, a residual dissection was noted at proximal LAD. IVUS examination also showed encircling gap that was noted between stent and vessel wall at proximal LAD stent area. Because distal flow was good and there was no ischemic symptom and sign, the patient was discharged. Six months later from index procedure, routine follow-up angiography and IVUS examination were performed and revealed more progressed previous residual coronary dissection at proximal LAD which was extended to bifurcation site. Our case showed, although intracoronary stenting might be an attractive approach by closure of the inlet and the false lumen, complete resolution of dissection by stenting is very important for long-term prognosis. PMID- 17397948 TI - Predictive models of syncope causes in an outpatient clinic. AB - The investigation of unexplained syncope remains a challenging clinical problem. In the present study we sought to evaluate the diagnostic value of a standardized work-up focusing on non invasive tests in patients with unexplained syncope referred to a syncope clinic, and whether certain combinations of clinical parameters are characteristic of rhythmic and reflex causes of syncope. METHODS AND RESULTS: 317 consecutive patients underwent a standardized work-up including a 12-lead ECG, physical examination, detailed history with screening for syncope related symptoms using a structured questionnaire followed by carotid sinus massage (CSM), and head-up tilt test. Invasive testings including an electrophysiological study and implantation of a loop recorder were only performed in those with structural heart disease or traumatic syncope. Our work up identified an etiology in 81% of the patients. Importantly, three quarters of the causes were established non invasively combining head-up tilt test, CSM and hyperventilation testing. Invasive tests yielded an additional 7% of diagnoses. Logistic analysis identified age and number of significant prodromes as the only predictive factors of rhythmic syncope. The same two factors, in addition to the duration of the ECG P-wave, were also predictive of vasovagal and psychogenic syncope. These factors, optimally combined in predictive models, showed a high negative and a modest positive predictive value. CONCLUSION: A standardized work up focusing on non invasive tests allows to establish more than three quarters of syncope causes. Predictive models based on simple clinical parameters may help to distinguish between rhythmic and other causes of syncope. PMID- 17397949 TI - Cholinergic stimulation with pyridostigmine prevents the impairment in ventricular function during mental stress in coronary artery disease patients. AB - Mental stress causes physiological autonomic adjustments that may trigger myocardial ischemia and ventricular dysfunction in patients with coronary artery disease. Thus, it was hypothetized that cholinergic stimulation may counteract the ventricular dysfunction provoked by mental stress in coronary disease. Six patients with coronary disease underwent a randomized, double-blind, cross-over, and placebo-controlled protocol in which they received placebo or a single dose of pyridostigmine bromide (45 mg p.o.), a reversible cholinesterase inhibitor, and thus, a cholinomimetic agent 2 h before a standard mental stress task (Stroop color-word test), while hemodynamic and echocardiographic variables were continuously monitored. There were no signs of myocardial ischemia on ECG during mental stress under PYR or placebo. Heart rate and blood pressure increased during mental stress (P<0.01) similarly with placebo and PYR (P>0.05). There were no ventricular wall motion abnormalities during mental stress with either placebo or PYR, but mental stress decreased ejection fraction (pre 63+/-2%, stress 57+/ 2%; P=0.004) and impaired the indices of diastolic ventricular function. On the other hand, PYR prevented the fall in ejection fraction (pre 62+/-2%, stress 64+/ 2%; P=0.13) and in the indices of diastolic function (P>0.05). In conclusion, cholinergic stimulation with pyridostigmine prevented the impairment in myocardial function during mental stress in patients with coronary artery disease. PMID- 17397950 TI - Acute myocardial infarction in developing countries: the importance of large national registries. PMID- 17397951 TI - Optimal wages in the market for nurses. PMID- 17397952 TI - Easy DNA extraction method and optimisation of PCR-Temporal Temperature Gel Electrophoresis to identify the predominant high and low GC-content bacteria from dairy products. AB - Molecular fingerprinting of bacterial ecosystems has recently increased in food microbiology. The aim of this work was to develop a rapid and easy method to extract DNA from various cheeses, and to optimize the separation of low and high GC-content bacteria by PCR-Temporal Temperature Gel Electrophoresis (PCR-TTGE). Seventy six strains belonging to 50 of the most frequently encountered bacterial species in dairy products were used to construct a database. Specific PCR-TTGE ladders containing 17 species forming a regular scale were created. Amplicons of these species were sequenced and the GC-content plotted against the migration distance: the correlation coefficients obtained were r(2)=0.97 and r(2)=0.99, respectively for high and low GC-contents. The extraction method developed did not use any harmful solvent such as phenol/chloroform. The concentrations of DNA extracted from hard cooked and pressed cheeses, quantified by picogreen molecular probes, were between 0.7 and 6 microg/g for core samples and 8 to 30 microg/g for rind samples. Experimental as well as commercial dairy products were analysed using the developed method and the reproducibility of the profiles was 89%. The method appears to be particularly efficient in the characterization of the ecosystem of cheese rinds. PMID- 17397953 TI - Differential analysis of protein expression of Bifidobacterium grown on different carbohydrates. AB - We observed recently that colonic fermentation of lactose might be a major factor in the pathophysiology of lactose intolerance. Proteomic techniques could be helpful in interpreting the metabolic pathways of lactose fermentation. The objective of this study was to explore proteomic methodologies for studying bacterial lactose metabolism that can be used to detect and identify proteins associated with the onset of intolerance symptoms. Differential expression of cytoplasmic proteins of Bifidobacterium animalis, Bifidobacterium breve and Bifidobacterium longum grown on different carbohydrates (lactose, glucose, galactose) was analyzed with surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) MS and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). After fractionation by SDS-PAGE, differentially expressed proteins were identified with LC-MS/MS. The three strains grown on the same carbohydrate or the same strain grown on glucose or lactose showed differences in SELDI-TOF MS protein profiles. Differences in protein expression were observed in B. breve grown on glucose, galactose or lactose as analyzed with SDS-PAGE. With LC-MS/MS, proteins from Bifidobacterium were identified, which included enzymes for metabolism of lactose, glucose and galactose. In conclusion, the applied techniques can discern differences in protein expression of bacteria metabolizing different carbohydrates. These techniques are promising in studying metabolism of lactose and other substrates in a complex bacterial ecosystem such as the colonic microbiota. PMID- 17397954 TI - An angiotensin II receptor antagonist suppresses running-enhanced hippocampal neurogenesis in rat. AB - Hippocampal neurogenesis is enhanced by voluntary running exercise in adult mammals. To elucidate the factors involved in this enhancement, we examined the effects of losartan, an antagonist of angiotensin II type 1 receptors, on the running-enhanced neurogenesis in the adult rat hippocampus. When losartan was administered orally via the drinking water, the running-enhanced cell proliferation in the subgranular zone was almost completely suppressed, indicating that this enhancement may be mediated by angiotensin II and its receptors. PMID- 17397955 TI - Microbial risk assessment of staphylococcal food poisoning in Korean kimbab. AB - The objective of this study was to conduct a preliminary microbial risk assessment for Staphylococcus aureus in kimbab, and then propose appropriate guidelines for its preparation and storage. Kimbab is a ready-to-eat product, popular in Korea, made of various foods surrounded by rice and seaweed. Because it is usually prepared by hand and stored at room temperature, it can be contaminated with S. aureus. Data on the prevalence, concentration and growth of S. aureus in kimbab as well as typical ambient temperatures in Seoul were collected; probability distributions were then selected to describe these data. A Monte Carlo simulation model was created using @risk. When current Korean retail data on S. aureus prevalence and concentration were used as inputs, the simulation predicted that the S. aureus concentration would be between 4.0 and 4.9 log CFU/g after 1 h of storage at ambient temperature during the warmest part of the year (March through November). Conversely, by assuming a starting concentration of 1 cell/g, the simulation predicted that the S. aureus concentration would reach 4.4 log CFU/g after 5 h of ambient storage in May and 3.4 log CFU/g in October. Our results suggest that, given the current prevalence and concentration of S. aureus found in kimbab at retail outlets, the product should be consumed within 1 h of purchase. Our results also indicate that if Korean consumers wish to safely store kimbab for 5 h at ambient temperature, S. aureus concentration should not exceed 1 CFU/g at the time of preparation. PMID- 17397956 TI - Calorimetric bioprocess monitoring by small modifications to a standard bench scale bioreactor. AB - Heat released during whole cell bioconversions provides information about the stoichiometry and kinetics of growth and product formation. It is considered as a potent control variable because it reflects immediately changes in metabolism. Furthermore, the sensitivity of calorimetric measurements is improving with increasing ratios of heat producing volume to heat exchanging surface and therefore with any scaling-up. However, the development and testing of calorimetry-based control strategies at the bench scale is difficult. Common microcalorimeters are unsuited due to their high price, the reactor volume (1 100mL) and the reactor design. Reaction calorimeters are even more expensive, less sensitive than microcalorimeters and often not adapted to bioprocesses. This paper describes a way-out by integrating calorimetric measurement principles into bench-scale standard bioreactors. The proposed solution benefits from all the measurement and control features of bench-scale bioreactors for, e.g. aeration, stirring, pH-control and substrate dosage. The additional calorimetric option can be easily constructed and integrated into an existing bioreactor at relatively low cost. The achieved sensitivity of 50 mW L(-1), stability of 0.2 mW L(-1)h(-1) and response time of a 1-2 min is in the same magnitude or even better than those of established reaction calorimeters. The calorimetrically monitored growth of the halophilic strain Halomonas elongata DMSZ 2581(T) demonstrates the applicability of the proposed solution for bioprocess analysis and control. PMID- 17397957 TI - Molecular systems biology. Proceedings of an international workshop. June 2006. Bielefeld, Germany. PMID- 17397958 TI - Evaluating the 3C-like protease activity of SARS-Coronavirus: recommendations for standardized assays for drug discovery. AB - Although the initial outbreaks of the deadly coronavirus that causes severe acute respiratory syndrome (SARS-CoV) were controlled by public health measures, the development of vaccines and antiviral agents for SARS-CoV is essential for improving control and treatment of future outbreaks. One potential target for SARS-CoV antiviral drug development is the 3C-like protease (3CLpro). This enzyme is an attractive target since it is essential for viral replication, and since there are now a number of high resolution X-ray structures of SARS-CoV 3CLpro available making structure-based drug-design possible. As a result, SARS-CoV 3CLpro has become the focus of numerous drug discovery efforts worldwide, but as a consequence, a variety of different 3CLpro expression constructs and kinetic assays have been independently developed making evaluation and comparison between potential inhibitors problematic. Here, we review the literature focusing on different SARS-CoV 3CLpro expression constructs and assays used to measure enzymatic activity. Moreover, we provide experimental evidence showing that the activity of 3CLpro enzymatic is significantly reduced when non-native sequences or affinity-tags are added to the N- or C-termini of the enzyme, or when the enzyme used in assays is at concentrations below the equilibrium dissociation constant of the 3CLpro dimer. We demonstrate for the first time the utility of a highly sensitive and novel Alexa488-QSY7 FRET-based peptide substrate designed for routine analysis and high-throughput screening, and show that kinetic constants determined from FRET-based assays that are uncorrected for inner-filter effects can lead to artifacts. Finally, we evaluated the effects of common assay components including DTT, NaCl, EDTA and DMSO on enzymatic activity, and we recommend standardized assay conditions and constructs for routine SARS-CoV 3CLpro assays to facilitate direct comparisons between SARS-CoV 3CLpro inhibitors under development worldwide. PMID- 17397959 TI - SARS coronavirus replicase proteins in pathogenesis. AB - Much progress has been made in understanding the role of structural and accessory proteins in the pathogenesis of severe acute respiratory syndrome coronavirus (SARS-CoV) infections. The SARS epidemic also brought new attention to the proteins translated from ORF1a and ORF1b of the input genome RNA, also known as the replicase/transcriptase gene. Evidence for change within the ORF1ab coding sequence during the SARS epidemic, as well as evidence from studies with other coronaviruses, indicates that it is likely that the ORF1ab proteins play roles in virus pathogenesis distinct from or in addition to functions directly involved in viral replication. Recent reverse genetic studies have confirmed that proteins of ORF1ab may be involved in cellular signaling and modification of cellular gene expression, as well as virulence by mechanisms yet to be determined. Thus, the evolution of the ORF1ab proteins may be determined as much by issues of host range and virulence as they are by specific requirements for intracellular replication. PMID- 17397960 TI - Delivery of G3139 using releasable PEG-linkers: impact on pharmacokinetic profile and anti-tumor efficacy. AB - In order to overcome the problems of enzymatic degradation and short plasma half life, which can limit the delivery of antisense oligonucleotides, and the potential immuno-stimulatory effects of CpG motifs, we utilized a polyethylene glycol (PEG) technology that employed various releasable linkers (rPEG). 5'-20 kDa-PEGylation of an anti-Bcl-2 5'-aminoalkyl-oligonucleotide with the same sequence as G3139 (Compound 1) did not alter its binding to the heparin-binding protein bFGF, nor the release of cytochrome c from isolated mitochondria treated with the conjugates. However, in 518A2 melanoma cells in vitro, PEGylation resulted in greatly diminished cellular uptake. In striking contrast, PEGylation of 1 resulted in dramatically improved pharmacokinetic profiles in vivo, with a prolonged half-life (t1/2), increased plasma concentration, and increased area under the plasma concentration-time curve (AUC). In an in vivo melanoma 518A2 xenograft mouse model, treatment with either 5'-20 kDa-PEG-1 or 1 demonstrated similar tumor growth inhibition. Furthermore, in an in vitro mouse splenocyte culture system, attachment of a PEG moiety to 1 through releasable linkers abolished the immunostimulatory response that was observed for G3139. Our results demonstrate the potential of the in vivo use of PEGylated oligonucleotides, and point out the profound differences between in vitro and in vivo models of oligonucleotide activity. PMID- 17397961 TI - Pro-fibrogenic potential of PDGF-D in liver fibrosis. AB - BACKGROUND/AIMS: We analyzed the expression of platelet-derived growth factor D (PDGF-D) in an experimental bile duct-ligated (BDL) rat model and assessed its biological function in cultured hepatic stellate cells (HSC) and myofibroblasts (MFB). METHODS: The mRNA for PDGF-A, -B, -C, -D and for PDGF receptor-alpha and beta chains (PDGFRalpha and PDGFRbeta) in normal and fibrotic rat livers was assessed quantitatively. Protein levels of PDGF-D were quantified by immunoblotting and immunohistochemistry. RESULTS: The relative mRNA expression of all PDGF isoforms and receptors upregulated upon BDL and PDGF-A, -B and -D expression was significantly higher than that of PDGF-C. PDGF-D and PDGFRbeta protein also increased markedly. Immunostaining revealed that PDGF-D is localized along the fibrotic septa of the periportal- and perisinusoidal areas. Besides PDGF-B, PDGF-D is the second most potent PDGF isoform in PDGFRbeta signaling within HSC/MFB, evidenced by PDGFRbeta autophosphorylation and activation of the downstream signaling molecules ERK1/2-, JNK-, p38 MAPK, and PKB/Akt while PDGF-C effects were minimal. PDGF-D exerted mitogenic and fibrogenic effects in both cultured HSC and MFB comparable to PDGF-B but PDGF-A and -C showed only marginal fibrogenic effects. CONCLUSIONS: PDGF-D possesses potential pathogenetic properties for HSC activation and matrix remodeling in liver fibrosis. PMID- 17397962 TI - Long-term survival of a patient with small cell lung cancer associated with cancer-associated retinopathy. AB - Cancer-associated retinopathy (CAR) is a rare paraneoplastic disorder that is frequently found in patients with small cell lung cancer (SCLC); it is caused by autoantibody to the 23-kDa photoreceptor protein, recoverin. We report a 9-year survivor of SCLC after concurrent chemoradiotherapy. His anti-recoverin antibody remains positive. Long-term survival without SCLC recurrence might be related to an autoimmunity mechanism that causes CAR due to the presence of anti-recoverin antibody cross-reacting with retinal cells and tumor cells. The current literature review was conducted to evaluate the impact on overall survival according to anti-recoverin antibody status. PMID- 17397963 TI - Field-scale effective matrix diffusion coefficient for fractured rock: results from literature survey. AB - Matrix diffusion is an important mechanism for solute transport in fractured rock. We recently conducted a literature survey on the effective matrix diffusion coefficient, D(m)(e), a key parameter for describing matrix diffusion processes at the field scale. Forty field tracer tests at 15 fractured geologic sites were surveyed and selected for the study, based on data availability and quality. Field-scale D(m)(e) values were calculated, either directly using data reported in the literature, or by reanalyzing the corresponding field tracer tests. The reanalysis was conducted for the selected tracer tests using analytic or semi analytic solutions for tracer transport in linear, radial, or interwell flow fields. Surveyed data show that the scale factor of the effective matrix diffusion coefficient (defined as the ratio of D(m)(e) to the lab-scale matrix diffusion coefficient, D(m), of the same tracer) is generally larger than one, indicating that the effective matrix diffusion coefficient in the field is comparatively larger than the matrix diffusion coefficient at the rock-core scale. This larger value can be attributed to the many mass-transfer processes at different scales in naturally heterogeneous, fractured rock systems. Furthermore, we observed a moderate, on average trend toward systematic increase in the scale factor with observation scale. This trend suggests that the effective matrix diffusion coefficient is likely to be statistically scale-dependent. The scale factor value ranges from 0.5 to 884 for observation scales from 5 to 2000 m. At a given scale, the scale factor varies by two orders of magnitude, reflecting the influence of differing degrees of fractured rock heterogeneity at different geologic sites. In addition, the surveyed data indicate that field-scale longitudinal dispersivity generally increases with observation scale, which is consistent with previous studies. The scale-dependent field-scale matrix diffusion coefficient (and dispersivity) may have significant implications for assessing long-term, large-scale radionuclide and contaminant transport events in fractured rock, both for nuclear waste disposal and contaminant remediation. PMID- 17397964 TI - Identification of a keratin-associated protein with a putative role in vesicle transport. AB - Protection of skin against UV light requires a coordinated interaction between melanocytes and keratinocytes. Melanosomes are lysosome-related organelles that originate in melanocytes and are transferred into keratinocytes where they form a supranuclear cap. The mechanism responsible for melanosome transfer into keratinocytes and their intracellular distribution is poorly understood. Recently, we reported for the first time that loss-of-function mutations in the keratin K5 gene affect melanosome distribution in keratinocytes and results in a reticulate hyperpigmentation disorder, called Dowling-Degos disease. Here, we characterise the distribution and behaviour of individual K5 and K14 domains following transient and stable transfection into cells. We report that the K5 head domain is considerably more stable than the K14 head. Moreover, the distribution of the K5 head domain is altered following depolymerisation of microtubules. Following co-immunoprecipitation, we verified a specific interaction between the head domain of K5 with Hsc70, a chaperone also involved in vesicle uncoating. We hypothesise that this interaction is involved in melanosome formation or transport in keratinocytes. Alternatively, it may have a general function in the regulation of keratin assembly. PMID- 17397965 TI - Localization and regulation of plasma membrane Ca(2+)-ATPase in bovine spermatozoa. AB - Calcium (Ca(2+)) signals, produced by the opening of plasma membrane entry channels, regulate a number of functions in spermatozoa such as capacitation and motility. The mechanisms of Ca(2+) removal from the sperm, required to restore resting [Ca(2+)](i), include plasma membrane Ca(2+)-dependent ATPase (PMCA) isoenzymes as well as a plasma membrane Na(+)-Ca(2+) exchanger. We have recently shown that bovine sperm PMCA is stimulated by PDC-109, a secretory protein of bovine seminal vesicles. To demonstrate the subcellular localization and regulation of bovine sperm PMCA, we have performed cell fractionation, enzyme activity determination and Western blotting studies of PMCA in spermatozoa removed from the cauda epididymidis of bull. Fractionation of sperm heads and tails resulted in a distinct association of ATPase activity with the tail membrane fraction. In vitro stimulation studies with PDC-109 using intact and fractionated sperm showed an increase in enzyme activity up to 105% in sperm tail membranes. Furthermore, thapsigargin inhibition did not alter the stimulatory effect of PDC-109 on ATPase activity, indicating that no sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA), but only PMCA isoenzymes are involved in this effect. Western blotting studies using a polyvalent PMCA antibody showed the exclusive presence of a 135 kDa band in the tail plasma membrane fraction. To elucidate whether or not the stimulatory effect was a direct one or indirectly mediated through PKA and PKC activation, PKA and PKC inhibitors, respectively, were used in the Ca(2+)-ATPase activity assays, which was followed by PDC-109 stimulation. The stimulatory effect of PDC-109 on PMCA was still observed under these conditions, while no phosphotyrosine proteins could be detected by Western blotting in sperm extracts following PDC-109 treatment. Co-immunoprecipitation studies, PDC-109 affinity chromatography as well as overlay blots failed to show a strong association of both PMCA and PDC-109, pointing to an indirect, perhaps phospholipid-mediated effect. PMID- 17397966 TI - Specific distribution of barrier-relevant ceramides in the emerging epidermis and the periderm/subperiderm during chicken embryogenesis. AB - During mammalian embryogenesis the emerging epidermis is temporarily covered by an epithelial monolayer, the periderm. In chicken, a second epithelial layer, the subperiderm, located underneath the periderm develops in later embryogenesis. Together the periderm and the subperiderm are referred to as the PSP unit. The cells of the PSP unit are tightly connected by tight junctions (TJ), thereby providing the embryo with an impermeable bilayered diffusion barrier. The emerging epidermis assumes its barrier function by cornification beginning at embryonic day 17 (E17) before at E18 the PSP unit undergoes desquamation. Lipid analysis of both epithelia after their mechanical separation revealed a dramatic increase to about 100-fold values of barrier-relevant ceramides, i.e. those known to essentially contribute to the diffusion barrier of the cornified envelope, in the emerging epidermis between E17 and E19. In contrast, the content of barrier relevant ceramides in the PSP unit remained at constantly low levels throughout embryogenesis. These data strongly argue in favour of different mechanisms for the barrier function of the two epithelia. TJ in the PSP unit provide the main diffusion barrier protecting the embryo until beginning of desquamation at E18. At this developmental stage the content of cornified envelope-specific ceramides is substantially elevated, thus enabling the epidermis to fulfil its function as the major diffusion barrier after desquamation of the PSP unit. The observation that barrier-relevant ceramides are formed prior to desquamation of the PSP unit points to a precisely regulated sequence in that desquamation does not occur until the lipid-based barrier of the cornified envelope is completed and suggests in addition that these lipids might be essential regulators of the interaction between the PSP unit and the emerging epidermis. PMID- 17397967 TI - Health status among internally displaced persons in Louisiana and Mississippi travel trailer parks. AB - STUDY OBJECTIVE: We used a global humanitarian aid perspective to assess basic needs, women's health, mental health, and opinions about the status of internally displaced persons living in travel trailer parks to inform recovery efforts for this population. METHODS: This was a systematic randomized survey of 366 internally displaced persons, conducted with structured questionnaires. The study setting was commercial and group travel trailer parks in Louisiana and Mississippi. Information was gathered about respondent demographics, food security, basic needs, domestic and sexual violence, security concerns, reproductive health, mental health, morbidity, mortality, health care assessment, substance use, and opinions about internally displaced persons and social status. RESULTS: Respondents were 45.9 (standard deviation 0.8) years of age on average and were mostly white (62%) in Mississippi and mostly black (65%) in Louisiana. Shelter, transportation, security, and lack of financial means were listed as the worst problems since displacement. Sixteen percent of respondents reported not having enough drinking water, and only 13% of those living in counties and parishes under boil orders were doing so. More than half of households reported an ill adult or child in the previous 2 months. The number of parents reporting problems getting children to school more than tripled after displacement. Intimate partner violence rates postdisplacement were 3 times higher than US baseline rates. Fifty percent of respondents met criteria for major depression. Suicide completion rates after displacement were more than 14 times the baseline rates, and attempt rates were more than 78 times baseline. CONCLUSION: The health burdens identified present a formidable challenge for the health infrastructures in Louisiana and Mississippi without outside assistance. Those planning and leading recovery efforts must understand internally displaced persons in a more global context and tailor programming that follows well-developed international models of rights-based care. PMID- 17397968 TI - Microarray analysis of oxidative stress regulated genes in mesencephalic dopaminergic neuronal cells: relevance to oxidative damage in Parkinson's disease. AB - Oxidative stress and apoptotic cell death have been implicated in the dopaminergic cell loss that characterizes Parkinson's disease. While factors contributing to apoptotic cell death are not well characterized, oxidative stress is known to activate an array of cell signaling molecules that participate in apoptotic cell death mechanisms. We investigated oxidative stress-induced cytotoxicity of hydrogen peroxide (H2O2) in three cell lines, the dopaminergic mesencephalon-derived N27 cell line, the GABAergic striatum-derived M213-20 cell line, and the hippocampal HN2-5 cell line. N27 cells were more sensitive to H2O2 induced cell death than M213-20 and HN2-5 cells. H2O2 induced significantly greater increases in caspase-3 activity in N27 cells than in M213-20 cells. H2O2 induced apoptotic cell death in N27 cells was mediated by caspase-3-dependent proteolytic activation of PKCdelta. Gene expression microarrays were employed to examine the specific transcriptional changes in N27 cells exposed to 100 microM H2O2 for 4 h. Changes in genes encoding pro- or anti-apoptotic proteins included up-regulation of BIK, PAWR, STAT5B, NPAS2, Jun B, MEK4, CCT7, PPP3CC, and PSDM3, while key down-regulated genes included BNIP3, NPTXR, RAGA, STK6, YWHAH, and MAP2K1. Overall, the changes indicate a modulation of transcriptional activity, chaperone activity, kinase activity, and apoptotic activity that appears highly specific, coordinated and relevant to cell survival. Utilizing this in vitro model to identify novel oxidative stress-regulated genes may be useful in unraveling the molecular mechanisms underlying dopaminergic degeneration in Parkinson's disease. PMID- 17397970 TI - [Post-transfusion acute lung injury (Trali) after plasma infusion in a patient having a constitutional thrombotic microangiopathy]. AB - INTRODUCTION: Transfusion-related acute lung injury is a post-transfusion interstitial lung injury. CASE REPORT: We reported a post-transfusion acute lung injury in a 23-years old woman having a chronic thrombotic microangiopathy related to an ADAMTS 13 constitutional deficiency receiving monthly plasma infusion for six years. The temporal relationship between the lung injury and the infusion of fresh frozen plasma led to the diagnosis of transfusion-related acute lung injury. The finding in the donor of the transfused plasma of an anti-HLA class II antibody recognizing HLA-DR52 present on leucocytes of the recipient suggests a causal relationship between this antigen-antibody conflict and the triggering of the TRALI. This chronic pathologic state requiring monthly plasma transfusions for thrombotic accident prevention raises the question of the selection of plasma obtained from non-immunized donors. CONCLUSION: The occurrence of a post transfusion pulmonary edema without cardio-vascular overload, must lead to consider a TRALI especially in predisposing clinical situations. In the case reported the role of constitutional ADAMTS 13 deficiency in genesis of TRALI is considered. PMID- 17397971 TI - [Adenosine deaminase is useful for the diagnosis of peritoneal tuberculosis in patients with end-stage renal failure]. AB - INTRODUCTION: End-stage renal failure patients are particularly at risk for tuberculosis, especially for peritoneal tuberculosis. Microbiological diagnosis remains hazardous in many cases. CASE REPORT: We report on a case of peritoneal tuberculosis in an end-stage renal failure patient. The diagnosis was suspected on the basis of adenosine deaminase dosage in peritoneal fluid, allowing an early presumptive treatment and a favourable outcome with a 3 years follow-up. DISCUSSION: The measurement of adenosine deaminase activity in ascites represents a diagnostic advance in tuberculous peritonitis among end-stage renal failure patients. PMID- 17397969 TI - The mitochondrial permeability transition in neurologic disease. AB - Mitochondria, being the principal source of cellular energy, are vital for cell life. Yet, ironically, they are also major mediators of cell death, either by necrosis or apoptosis. One means by which these adverse effects occur is through the mitochondrial permeability transition (mPT) whereby the inner mitochondrial membrane suddenly becomes excessively permeable to ions and other solutes, resulting in a collapse of the inner membrane potential, ultimately leading to energy failure and cell necrosis. The mPT may also bring about the release of various factors known to cause apoptotic cell death. The principal factors leading to the mPT are elevated levels of intracellular Ca2+ and oxidative stress. Characteristically, the mPT is inhibited by cyclosporin A. This article will briefly discuss the concept of the mPT, its molecular composition, its inducers and regulators, agents that influence its activity and describe the consequences of its induction. Lastly, we will review its potential contribution to acute neurological disorders, including ischemia, trauma, and toxic-metabolic conditions, as well as its role in chronic neurodegenerative conditions such as Alzheimer's disease, Parkinson's disease, Huntington's disease and amyotrophic lateral sclerosis. PMID- 17397972 TI - [Pelvic pain and inability to walk after seizure]. PMID- 17397973 TI - Learning to let go: the challenge of de-crowding the curriculum. AB - Nurse educators often lament there is not enough time to teach all the content students need to learn. Every year, information regarding healthcare increases exponentially. Typically, educators respond by increasing content without removing outdated information resulting in overcrowded courses. Little time is left for meaningful learning of concepts and life-long learning skills to sustain graduates throughout their careers. Reliance on conventional teacher-centered approaches to curriculum development have generated calls from professional organizations for innovative education programs. Innovation begins by shifting focus from massive amounts of content to teaching essential concepts and abilities necessary for today's health care environment. Decreasing the overload of content is imperative when moving from a teaching centered to a learning centered curricula. Instead of concentrating on rote memorization, students engage in meaningful learning. De-crowding the curriculum can be frustrating because there is no quick fix. The challenge is to consider both faculty philosophical beliefs and assumptions about how students learn. Assumptions and beliefs regarding teaching, learning, essential content, and the learning environment should be clear from the outset. Without considering assumptions and beliefs efforts to de-crowd the curriculum rapidly devolve into a test of wills between faculty members. The authors present an overview of learning-centered education, a systematic method for de-crowding the curriculum, and a discussion of challenges encountered. PMID- 17397974 TI - Some aspects of the recombinantly expressed humanised superagonist anti-CD28 mAb, TGN1412 trial catastrophe lessons to safeguard mAbs and vaccine trials. AB - We consider essential, still ignored, basic research aspects of the failed clinical trial (13 March 2006) of a recombinantly expressed humanised superagonist anti-CD28 mAb, TGN14122. Without hindsight, if for approval of the first ever recombinantly expressed anti-CD28 mAb use in humans attention had been paid to the physico-chemical factors and receptor saturation, the possible catastrophe will have been predictable and preventable. To understand what went wrong and, crucially, to prevent any future disasters to safeguard human health, safety and welfare, the information provided is likely to be of wide interest. We present calculations to show CD28 receptors on T cells of the six healthy volunteers by the anti-CD28 mAb superagonist, TGN1412. This led to the over activation of T cells and the violent cytokine storm precipitating the cascade and the release of endogenous molecules affecting other cells. Monocytes and plasma cells are likely to have been affected. We discuss briefly the role of neutrophils and activation releasing the surface-located sialidase affecting cell coats, such as, of T lymphocytes exposing galactose receptors that could have been involved in antigen presenting cell interactions. The role of the cell surface thiols of lymphocytes in forming mixed disulphides with endogenous ligands and in the REDOX system are briefly mentioned. Consideration of these various factors and a critical evaluation of the receptor occupancy data before injecting 0.1 mg/kg TGN1412 will have rung alarm bells about possible serious side effects and the catastrophe will have been averted. PMID- 17397975 TI - 'Creating a safety net': Women's experiences of antenatal depression and their identification of helpful community support and services during pregnancy. AB - OBJECTIVE: to explore the feelings of depression during pregnancy of a local sample of women living in an area of socio-economic deprivation, and to identify the support mechanisms that they report as personally or potentially helpful for antenatal depression. DESIGN: a retrospective study using a qualitative approach, informed by constructivism, to explore the participants' individual experiences of depression during pregnancy. Data were collected via tape-recorded semi structured interviews. SETTING: a socio-economically deprived area in North London, UK, identified as a Sure Start Local Programme providing local services specifically designed for socially disadvantaged families with children aged 0-4 years. PARTICIPANTS: a self-selected sample of nine women aged 23-40 years, from a variety of ethnic backgrounds, who retrospectively admitted to feeling low or depressed during pregnancy. All the participants had had a baby more than 6 weeks previously and less than 1 year before the start of the study. FINDINGS: despite different cultural and ethnic backgrounds, the participants shared similar feelings of emotional isolation that seemed to contribute largely to their experience of antenatal depression. Partner support (or lack of it) seemed to be crucial to the women's psychological well-being during pregnancy. For some of these women, the research interview was the first opportunity to talk about their needs and feelings during pregnancy. Potentially helpful mechanisms for support were identified by the participants and were judged to be relatively simple to introduce, involving connecting with other women via peer support and having 'somewhere to go' to meet others during pregnancy. IMPLICATIONS FOR PRACTICE: some women do not disclose their feelings of depression during pregnancy, with potentially damaging effects on both the family and the baby. Feelings of loss and emotional isolation may occur, which could be partly alleviated by providing models of midwifery care that offer continuity of carer. Isolated and vulnerable women require increased midwifery resources, and partners may also have particular needs for support and adjustment, which currently remain unmet and need further research. Many 'low tech' interventions aimed at supporting women with antenatal depression could be developed, including peer support, which may offer realistic models of social capital and community empowerment in the new Children's Centres in England and Wales. PMID- 17397976 TI - Childhood cognitive ability and adult mental health in the British 1946 birth cohort. AB - We examined whether childhood cognitive ability was associated with two mental health outcomes at age 53 years: the 28 item General Health Questionnaire (GHQ 28) as a measure of internalising symptoms of anxiety and depression, and the CAGE screen for potential alcohol abuse as an externalising disorder. A total of 1875 participants were included from the Medical Research Council National Survey of Health and Development, also known as the British 1946 birth cohort. The results indicated that higher childhood cognitive ability was associated with reporting fewer symptoms of anxiety and depression GHQ-28 scores in women, and increased risk of potential alcohol abuse in both men and women. Results were adjusted for educational attainment, early socioeconomic status (SES) and adverse circumstances, and adult SES, adverse circumstances, and negative health behaviours. After adjusting for childhood cognitive ability, greater educational attainment was associated with reporting greater symptoms of anxiety and depression on the GHQ-28. Although undoubtedly interrelated, our evidence on the diverging effects of childhood cognitive ability and educational attainment on anxiety and depression in mid-adulthood highlights the need for the two to be considered independently. While higher childhood cognitive ability is associated with fewer internalising symptoms of anxiety and depression in women, it places both men and women at higher risk for potential alcohol abuse. Further research is needed to examine possible psychosocial mechanisms that may be associated with both higher childhood cognitive ability and greater risk for alcohol abuse. In addition, the underlying mechanisms responsible for the gender-specific link between childhood cognitive ability and the risk of experiencing internalising disorders in mid-adulthood warrants further consideration. PMID- 17397977 TI - Education does not protect against age-related decline of switching focal attention in working memory. AB - In this experimental study, effects of age and education on switching focal attention in working memory were investigated among 44 young (20-30 years) and 40 middle-aged individuals (50-60 years). To this end, a numeric n-back task comprising two lag conditions (1- and 2-back) was administered within groups. The results revealed a comparable increase of reaction time as a function of lag across age groups, but a disproportionate decrease of accuracy in the middle-aged relative to the young group. The latter effect did not interact with education, which challenges the cognitive reserve hypothesis. Moreover, the high-educated middle-aged participants showed a greater increase of reaction time as a function of lag than their low-educated counterparts. Apparently, they were not able to sustain their relatively high response speed across conditions. These results suggest that education does not protect against age-related decline of switching focal attention in working memory. PMID- 17397978 TI - Too much to count on: impaired very small numbers in corticobasal degeneration. AB - Patients with corticobasal degeneration (CBD) have calculation impairments. This study examined whether impaired number knowledge depends on verbal mediation. We focused particularly on knowledge of very small numbers, where there is a precise relationship between a cardinality and its number concept, but little hypothesized role for verbal mediation. We evaluated accuracy and reaction time (RT) for matching dot arrays and Arabic numerals involving smaller (2-4) and larger (5-9) cardinalities in non-aphasic patients with CBD (n=16), frontotemporal dementia (FTD; n=23), and healthy controls (n=15). CBD were less accurate and slowed at judging smaller Arabic numeral-dot array stimuli compared to FTD patients and controls. Moreover, only CBD showed longer RTs judging successively larger number-dot array pairs among the smaller cardinalities. Difficulty judging very small numbers is impaired in CBD, suggesting degraded representation of precise number knowledge that does not depend on language functioning. PMID- 17397979 TI - Optokinetic stimulation affects temporal estimation in healthy humans. AB - The representation of time and space are closely linked in the cognitive system. Optokinetic stimulation modulates spatial attention in healthy subjects and patients with spatial neglect. In order to evaluate whether optokinetic stimulation could influence time perception, a group of healthy subjects performed "time-comparison" tasks of sub- and supra-second intervals before and after leftward or rightward optokinetic stimulation. Subjective time perception was biased by the direction of optokinetic stimulation. Rightward optokinetic stimulation induced an overestimation of time perception compared with baseline and leftward optokinetic stimulation. These results indicate a directional bias in time perception induced by manipulation of spatial attention and could argue for a mental linear representation of time intervals. PMID- 17397980 TI - Safety evaluation of a milk basic protein fraction. AB - Milk products are widely consumed by individuals in the US population in the form of fluid milk and milk-derived products and ingredients. Milk is a good source of calcium, which plays a role in maintaining bone health. In addition to calcium, the whey protein fraction of milk contains basic proteins that have been demonstrated to increase bone metabolism and inhibit bone resorption. A specific basic protein fraction in milk (Milk Basic Protein; MBP) was tested in an acute oral toxicity study, teratology study, subchronic oral toxicity study, and reverse mutation assay and no treatment related adverse effects were found. MBP has been evaluated for its use as an ingredient in food and concluded to be safe for its intended use. PMID- 17397981 TI - Repeated administration of diphenyl ditelluride induces hematological disorders in rats. AB - In the present study, we investigated potential toxic effects of diphenyl ditelluride, as measured by biochemical and hematological parameters. Rats were given a daily dose of 0.3 micromol/kg diphenyl ditelluride by subcutaneous route and sacrificed at different times (24 and 48 h). Hepatic and renal TBARS levels were changed by diphenyl ditelluride exposure at the dose 0.9 micromol/Kg in rats. Diphenyl ditelluride exposure demonstrated an increase in AST (aspartate aminotransferase), ALT (alanine aminotransferase) and LDH (lactate dehydrogenase) activities. Plasma creatinine and urea levels increase after diphenyl ditelluride exposure. Diphenyl ditelluride also produced a significant decrease in plasma triglyceride and cholesterol levels. In contrast, this compound, at all doses tested, induced a marked increase in total leukocyte counts. The present study suggests that diphenyl ditelluride induces hematological disorders and provides evidence for renal and hepatic toxicity in rats. PMID- 17397982 TI - First European conference on aspartame: putting safety and benefits into perspective. Synopsis of presentations and conclusions. AB - A Conference was held in Paris in 2006 to review the safety and benefits arising from the replacement of sucrose with the intense sweetener aspartame. The intakes of aspartame are only about 10% of the acceptable daily intake, even by high consumers, so that the safety margin is about 3 orders of magnitude. The safety of aspartame was confirmed in the EFSA Opinion of a recent controversial rodent cancer bioassay. There is increasing evidence that even modest reductions in the intake of calories can reduce the risk factors associated with a number of diseases, such as diabetes and cardiovascular disease. A key issue addressed at the conference was whether the replacement of sucrose with aspartame could result in a prolonged decrease in calorie intake that was of similar magnitude to that necessary to produce a health benefit. A recent meta-analysis of published data showed that an adequate, prolonged weight reduction could be achieved with aspartame. It was recognised that risk assessment alone gave an unbalanced impression to regulators and consumers, and that in the future quantitative risk benefit analyses should be able to provide more comprehensive advice. PMID- 17397983 TI - New insights into the membrane topology of the phagocyte NADPH oxidase: characterization of an anti-gp91-phox conformational monoclonal antibody. AB - Cytochrome b(558) is the catalytic core of the phagocyte NADPH oxidase that mediates the production of bactericidal reactive oxygen species. Cytochrome b(558) is formed by two subunits gp91-phox and p22-phox (1/1), non-covalently associated. Its activation depends on the interaction with cytosolic regulatory proteins (p67-phox, p47-phox, p40-phox and Rac) leading to an electron transfer from NADPH to molecular oxygen and to the release of superoxide anions. Several studies have suggested that the activation process was linked to a change in cytochrome b(558) conformation. Recently, we confirmed this hypothesis by isolating cytochrome b(558) in a constitutively active form. To characterize active and inactive cytochrome b(558) conformations, we produced four novel monoclonal antibodies (7A2, 13B6, 15B12 and 8G11) raised against a mixture of cytochrome b(558) purified from both resting and stimulated neutrophils. The four antibodies labeled gp91-phox and bound to both native and denatured cytochrome b(558). Interestingly, they were specific of extracellular domains of the protein. Phage display mapping combined to the study of recombinant gp91-phox truncated forms allowed the identification of epitope regions. These antibodies were then employed to investigate the NADPH oxidase activation process. In particular, they were shown to inhibit almost completely the NADPH oxidase activity reconstituted in vitro with membrane and cytosol. Moreover, flow cytometry analysis and confocal microscopy performed on stimulated neutrophils pointed out the capacity of the monoclonal antibody 13B6 to bind preferentially to the active form of cytochrome b(558). All these data suggested that the four novel antibodies are potentially powerful tools to detect the expression of cytochrome b(558) in intact cells and to analyze its membrane topology. Moreover, the antibody 13B6 may be conformationally sensitive and used as a probe for identifying the active NADPH oxidase complex in vivo. PMID- 17397984 TI - Epidermal growth factor stimulates matrix metalloproteinase-9 expression and invasion in human follicular thyroid carcinoma cells through Focal adhesion kinase. AB - In order to further advance our knowledge of the role epidermal growth factor (EGF) plays in thyroid carcinoma, we investigated its effect on the regulation of matrix metalloproteinase-9 (MMP-9), a key enzyme that plays an important role in tumor invasion and angiogenesis. The expression of MMP-9 in EGF-treated and untreated human follicular thyroid carcinoma cells (FTC-133) was evaluated using reverse transcription-PCR, Western blot and gelatin zymography. Transient transfection and electrophoretic mobility shift assays (EMSA) were also performed to measure MMP-9 promoter activity, to identify multiple signaling pathways and to determine a proximal AP-1-binding site located between -79 to -73 base pairs upstream of the transcriptional start site that is involved in activation of MMP 9 by EGF. In the present study, we demonstrate that EGF treatment up-regulated MMP-9 expression in human follicular thyroid carcinoma cells. Expression of FAK related non kinase (FRNK), a potent dominant-negative inhibitor of FAK, reduced FAK auto-phosphorylation and inhibited EGF-induced MMP-9 transcription and secretion leading to decreased cell invasion through Matrigel in in vitro Transwell assays. Our studies highlight the role FAK plays in promoting cell invasion through the activation of distinct signaling pathways induced by EGF with protein MMP-9 transcription and secretion in follicular thyroid carcinoma cells. PMID- 17397985 TI - Thirty years of Escherichia coli DNA gyrase: from in vivo function to single molecule mechanism. AB - The level of negative DNA supercoiling of the Escherichia coli chromosome is tightly regulated in the cell and influences many DNA metabolic processes including DNA replication, transcription, repair and recombination. Gyrase is the only type II topoisomerase able to introduce negative supercoils into DNA, a unique ability that arises from the specialized C-terminal DNA wrapping domain of the GyrA subunit. Here, we review the biological roles of gyrase in vivo and its mechanism in vitro. PMID- 17397986 TI - Does topoisomerase II specifically recognize and cleave hairpins, cruciforms and crossovers of DNA? AB - DNA topoisomerase II is an enzyme that specializes in DNA disentanglement. It catalyzes the interconversion of DNA between different topological states. This event requires the passage of one duplex through another one via a transient double-strand break. Topoisomerase II is able to process any type of DNA, including structures such as DNA juxtapositions (crossovers), DNA hairpins or cruciforms, which are recognized with high specificity. In this review, we focused our attention on topoisomerase II recognizing DNA substrates that possess particular geometries. A strong cleavage site, as we identified in pBR322 DNA in the presence of ellipticine (site 22), appears to be characterized by a cruciform structure formed from two stable hairpins. The same sequence could also constitute a four-way junction structure stabilized by interactions involving ATC sequences. The latter have been shown to be able to promote Holliday junctions. We reviewed the recent literature that deals with the preferential recognition of crossovers by various topoisomerases. The single molecule relaxation experiments have demonstrated the differential abilities of the topoisomerases to recognize crossovers. It appears that enzymes, which distinguish the chirality of the crossovers, possess specialized domains dedicated to this function. We also stress that the formation of crossovers is dependent on the presence of adequate stabilizing sequences. Investigation of the impact of such structures on enzyme activity is important in order to both improve our knowledge of the mechanism of action of the topoisomerase II and to develop new inhibitors of this enzyme. PMID- 17397987 TI - cyclicAMP and glucocorticoid responsiveness of the rat carbamoylphosphate synthetase gene requires the interplay of upstream regulatory units. AB - Many genes involved in metabolic processes are regulated by glucocorticoids and/or cyclicAMP. The hepatic expression of the urea cycle enzyme carbamoylphosphate-synthetase-I gene (CPS) is regulated at the transcriptional level by both factors. Here, we report that the 5' half of the distal enhancer is necessary and sufficient for full cyclicAMP responsiveness. The cyclicAMP responsive element (CRE), and FoxA- and C/EBP-binding sites are indispensible for cyclicAMP responsiveness, indicating that these elements make up a cyclicAMP responsive unit (CRU). In addition to this CRU, the CPS regulatory regions contain two glucocorticoid-response elements (GRE): one in the 3' region of the distal enhancer and one in the proximal enhancer. In presence of the cyclicAMP responsive region in the distal enhancer, only one of the GREs is required for glucocorticoid-inducible CPS expression, with both GREs acting in an additive fashion to fully confer the inducing effect of glucocorticoids. In contrast, the simultaneous presence of both GREs is required in the absence of the cyclicAMP responsive region. In this configuration, the distal GRE fully depends on its neighbouring FoxA and C/EBP REs for activity and is, therefore, a glucocorticoid responsive unit. In conclusion, we show here that the CPS CRU is a bifunctional unit that elicits the cyclicAMP response and, in addition, functions as a glucocorticoid accessory unit to establish a glucocorticoid response from otherwise silent proximal or distal GRUs. Therefore, cyclicAMP and glucocorticoid pathways can induce CPS transcription via overlapping sets of response elements. PMID- 17397988 TI - Modulation of Bax Inhibitor-1 and cytosolic Ca2+ by cytokinins in Nicotiana tabacum cells. AB - The protein Bax Inhibitor-1 (BI-1) has recently emerged as a negative regulator of plant programmed cell death (PCD), but how it functions at the biochemical level remains unknown. To elucidate its regulation and mode of action, we used suspension cells of Nicotiana tabacum to study the effects of cytokinins (CKs) on the expression level of NtBI-1 via western analysis. We found that the NtBI-1 protein is up-regulated following treatments with CKs at concentrations inducing a stress response (determined by growth reduction and PR1a accumulation), but not at PCD-inducing concentrations. These data point toward a role for NtBI-1 in the stress response to CKs. Application of CKs was also accompanied by a rapid cytosolic Ca(2+) pulse, and inhibition of this pulse with La(3+) or EGTA partially restored viability, indicating a signaling role for Ca(2+) in CK induced cell death. However, CK-induced NtBI-1 accumulation was not altered by pretreatment with La(3+), nor by treatment with several modulators of intracellular Ca(2+) homeostasis and signaling, suggesting that CK-dependent regulation of NtBI-1 accumulation is not directly mediated by Ca(2+). PMID- 17397989 TI - Gradual melting of a replication origin (Schizosaccharomyces pombe ars1): in situ atomic force microscopy (AFM) analysis. AB - Local DNA melting is integral to fundamental processes such as replication or transcription. In vivo, these two processes do not occur on molecules free in solution but, instead, involve DNA molecules which are organized into DNA/proteins complexes. Atomic force microscopy imaging offers a possibility to look at individual molecules. It allowed us to follow the progress of local denaturation in liquid, but with the added constraints of DNA lying on a surface. We present a kinetic analysis of the mapping of the temperature-driven melting seen at a replication origin (Schizosaccharomyces pombe ars1). The results indicate an expected base composition dependency, but also a strong extremity effect. Noteworthy, a "structural" effect is clearly occurring - which is shown by the greater susceptibility of the strongly curved region present in the sequence to unwind. DNA melting, at this place, is seen to occur after an increase in the curvature amplitude and a simultaneous shift of the nucleotide sequence positioned at the apex. Because this may determine the position of the Replication Initiation (R.I.) site, the result suggests that eukaryotic replication origins, although described as possessing no consensus sequences, may well have their mechanics sustained by the properties of common structural features. Our analysis may, therefore, provide new information that will give genuine insights on how DNA molecules behave when organized into primosomes, replisomes, promoter initiation complexes, etc. and thus, be essential to better understanding the way genes function. PMID- 17397990 TI - Duration of dysmenorrhoea and extent of adenomyosis visualised by magnetic resonance imaging. AB - OBJECTIVE: Enlargement of the junctional zone (JZ) on T2-weighted resonance imaging of the uterus has recently been established as the major criterion for adenomyosis in patients with endometriosis. This study was conducted to analyse the extent of adenomyosis using magnetic resonance imaging (MRI) and relate it to the duration of dysmenorrhoea. STUDY DESIGN: This was a prospective study of 70 patients presenting with the complaint of severe dysmenorrhoea. Forty patients (57%) reported dysmenorrhoea as their major complaint and 30 patients (43%) suffered additionally from infertility. Group I (n=40) consisted of patients with dysmenorrhoea of between 1 and 10 years' duration, group II (n=30) consisted of patients with dysmenorrhoea of longer than 11 years' duration. All patients underwent laparoscopy to detect the presence and degree of endometriosis, and all patients underwent T2-weighted resonance imaging of the uterus to detect the extent of adenomyosis by measurement of the "junctional zone". RESULTS: In group I, adenomyosis could be detected via MRI in 21 patients (52.5%), while 19 patients (47.5%) showed no signs of adenomyosis. By contrast, in group II a distinct enlargement of the JZ, as the major radiological criterion of adenomyosis, could be observed in 26 patients (87%), while only 4 patients (13%) revealed no signs of adenomyosis (p=0.04). The mean thickness of the JZ was significantly enlarged in group II (11.07 mm) compared with group I (6.38 mm; p<0.0001). The prevalence of adenomyosis in endometriosis after dysmenorrhoea of more than 11 years' duration was 87%. CONCLUSIONS: In deep infiltrating endometriosis, a correlation between a specific localisation and dysmenorrhoea can often not be found. Recently, endometriosis and adenomyosis have been believed to result from a common uterine disease, the dislocation of the basal endometrium. Our data clearly show that dysmenorrhoea of long duration in patients who have had endometriosis for over a threshold value of 11 years is significantly related to adenomyosis of the uterus. Hence, evaluation of adenomyosis using MRI should become a standard procedure in cases of dysmenorrhoea and endometriosis. Severe dysmenorrhoea of long duration should always focus clinical interest on adenomyosis of the uterus. PMID- 17397991 TI - Prediction of stone-free rate after ESWL. PMID- 17397992 TI - Detection of lymph-node metastases with integrated [11C]choline PET/CT in patients with PSA failure after radical retropubic prostatectomy: results confirmed by open pelvic-retroperitoneal lymphadenectomy. AB - OBJECTIVES: To prospectively evaluate the accuracy of integrated [(11)C]choline PET/CT in the diagnosis of lymph-node recurrence in prostate cancer patients with biochemical failure after surgery. METHODS: Since October 2002, 25 patients with biochemical recurrence (median PSA: 1.98 ng/ml), based on evidence of lymph-node metastases on [(11)C]choline-PET/CT scan (21 cases) or conventional imaging (4 cases), were scheduled for either bilateral pelvic (12 cases) or both pelvic and retroperitoneal lymph-node dissection (13 patients). RESULTS: Sixty-three nodal sites were evaluated histologically. The mean number of nodes removed and positive nodes were 21.92+/-16.91 (range: 4-74) and 8.84+/-9.65 (range: 1-31), respectively. Of the four patients with negative [(11)C]choline-PET/CT and positive magnetic resonance, none had nodal metastases. Nineteen of the 21 patients (90%) with positive [(11)C]choline-PET/CT had nodal metastases of prostate adenocarcinoma at histologic evaluation. A lesion-based analysis showed that [(11)C]choline-PET/CT sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 64%, 90%, 86%, 72%, and 77%, respectively. The mean maximum diameter of true positive metastases was larger than false-negative ones (15.0 vs. 6.3mm; p=0.0004). CONCLUSIONS: [(11)C]Choline PET/CT is an accurate diagnostic tool for the detection of lymph-node metastases of recurrent prostate cancer. The low negative predictive value seems to depend on the limited capability of [(11)C]choline-PET/CT to detect microscopic lesions. The high positive predictive value, even with low PSA values, provides a basis for further treatment decisions. PMID- 17397993 TI - Editorial comment on: Detection of lymph-node metastases with integrated [11C]choline PET/CT in patients with PSA failure after radical prostatectomy: results confirmed by open pelvic-retroperitoneal lymphadenectomy. PMID- 17397994 TI - Does CSF hypocretin-1 decrease in Bickerstaff's brainstem encephalitis? PMID- 17397995 TI - Do skull base lesions and their surgical treatment cause epileptic seizures? AB - OBJECTIVES: Few reports exist on the risk of seizures with skull base lesions and their surgical treatment. PATIENTS AND METHODS: All patients referred for surgery of a skull base lesion at George Washington University Hospital from January 1998 to August 1999 were reviewed. After excluding patients with other risk factors 136 patients were included. RESULTS: No seizures were reported before or after surgery, but 59 patients (43%) were taking anticonvulsants. Only 17 (12.5%) had EEG. Four of these showed no abnormalities, 2 were abnormal without sharp waves and 11 had sharp waves (an increase compared to the general population). Most sharp waves affected the temporal lobe on the side of surgery. All EEGs were performed less than 4 months after surgery. CONCLUSION: This study does not indicate that seizures are common in this patient group, but there was an increased amount of epileptiform activity on EEG and many patients were taking AED. A prospective study with systematic patient interviews, pre- and post surgical EEG and a longer follow up is needed. PMID- 17397996 TI - Combined removal of BTEX in air stream by using mixture of sugar cane bagasse, compost and GAC as biofilter media. AB - Biofiltration of air stream containing mixture of benzene, toluene, ethyl benzene and o-xylene (BTEX) has been studied in a lab-scale biofilter packed with a mixture of compost, sugar cane bagasse and granulated activated carbon (GAC) in the ratio 55:30:15 by weight. Microbial acclimation was achieved in 30 days by exposing the system to average BTEX inlet concentration of 0.4194 gm(-3) at an empty bed residence time (EBRT) of 2.3 min. Biofilter achieved maximum removal efficiency more than 99% of all four compounds for throughout its operation at an EBRT of 2.3 min for an inlet concentration of 0.681 gm(-3), which is quite significance than the values reported in the literature. The results indicate that when the influent BTEX loadings were less than 68 gm(-3)h(-1) in the biofilter, nearly 100% removal could be achieved. A maximum elimination capacity (EC) of 83.65 gm(-3)h(-1) of the biofilter was obtained at inlet BTEX load of 126.5 gm(-3)h(-1) in phase IV. Elimination capacities of BTEX increased with the increase in influent VOC loading, but an opposite trend was observed for the removal efficiency. The production of CO(2) in each phase (gm(-3)h(-1)) was also observed at steady state (i.e. at maximum removal efficiency). Moreover, the high concentrations of nitrogen in the nutrient solution may adversely affect the microbial activity possibly due to the presence of high salt concentrations. Furthermore, an attempt was also made to isolate the most profusely grown BTEX degrading strain. A Gram-positive strain had a high BTEX-degrading activity and was identified as Bacillus sphaericus by taxonomical analysis, biochemical tests and 16S rDNA gene analysis methods. PMID- 17397997 TI - Novel chitosan derivative for the removal of cadmium in the presence of cyanide from electroplating wastewater. AB - Chitosan was chemically modified by introducing xanthate group onto its backbone using carbondisulfide under alkaline conditions. The chemically modified chitosan flakes (CMC) was used as an adsorbent for the removal of cadmium ions from electroplating waste effluent under laboratory conditions. CMC was found to be far more efficient than the conventionally used adsorbent activated carbon. The maximum uptake of cadmium by CMC in batch studies was found to be 357.14 mg/g at an optimum pH of 8.0 whereas for plain chitosan flakes it was 85.47 mg/g. Since electroplating wastewater contains cyanide in appreciable concentrations, interference of cyanide ions in cadmium adsorption was found to be very significant. This problem could be easily overcome by using higher doses of CMC, however, activated carbon was not found to be effective even at higher doses. Due to the high formation constant of cadmium with xanthate and adsorption was carried out at pH 8, cations like Pb(II), Cu(II), Ni(II) and Zn(II) did not interfere in the adsorption. Dynamics of the sorption process were studied and the values of rate constant of adsorption were calculated. Desorption of the bound cadmium from CMC was accomplished with 0.01 N H(2)SO(4). The data from regeneration efficiencies for 10 cycles evidenced the reusability of CMC in the treatment of cadmium-laden wastewater. PMID- 17397998 TI - Tanshinone IIB, a primary active constituent from Salvia miltiorrhza, exhibits neuro-protective activity in experimentally stroked rats. AB - Tanshinone IIB (TSB) is a major active constituent of the root of Salvia miltiorrhiza (Danshen) used in the treatment of acute stroke. Danshen extracts and TSB have shown marked neuron-protective effects in mouse studies but there is a lack of clinical evidence for the neuron-protective effects of Danshen and its active ingredients. This study investigated the neuron-protective effects of TSB in experimentally stroked rats. TSB at 5 and 25 mg/kg by intraperitoneal injection significantly reduced the focal infarct volume, cerebral histological damage and apoptosis in rats subjected to middle cerebral artery occlusion (MCAO) compared to MCAO rats receiving vehicle. This study demonstrated that TSB was effective in reducing stroke-induced brain damage and may represent a novel drug candidate for further development. Further mechanistic studies are needed for the neuron-protective activity of TSB. PMID- 17397999 TI - Evidence of disturbed amygdalar energy metabolism in patients with borderline personality disorder. AB - In order to detect possible links between structural and neurochemical brain abnormalities we applied high resolution morphometric imaging and short-echo time absolute-quantification magnetic resonance spectroscopy (MRS) at the left hand side to the amygdala in 12 patients with borderline personality disorder (BPD) and 10 group-matched healthy controls. Confirming earlier reports we found a significant 11-17% reduction of amygdalar volumes in patients with BPD. In addition there was a significant 17% increase of left amygdalar creatine concentrations in BPD patients. Left amygdalar creatine concentration correlated positively with measures of anxiety and negatively with amygdalar volume. This pilot study of simultaneous amygdalar morphometry and spectroscopy in BPD reveals a possible link between amygdalar volume loss, psychopathology and neurochemical abnormalities in terms of creatine signals. PMID- 17398000 TI - Raloxifene blocks estradiol induction of the serotonin transporter and 5 hydroxytryptamine2A receptor in female rat brain. AB - Sex steroids have potent effects on mood, mental state and cognition. Our previous findings and those of others suggest that these effects may be due at least in part to estradiol actions on central serotonergic mechanisms. Specifically, estradiol-17beta in its acute positive feedback mode for gonadotropin release in the female rat induces expression of the genes for the 5 hydroxytryptamine(2A) receptor (5-HT(2A)R) and the serotonin transporter (SERT) in the dorsal raphe nucleus (DRN). This is accompanied by an increase in the densities of 5-HT(2A)R and the SERT in forebrain regions which in the human are concerned with the control of mood, mental state, cognition and emotion. Here we report that raloxifene, a benzothiophene and selective estrogen receptor modulator (SERM), completely blocked estradiol stimulation of brain 5-HT(2A)R and SERT expression in acutely ovariectomized rats. Raloxifene also blocked the estrogen-induced surge of luteinizing hormone. Treatment of acutely ovariectomized rats with raloxifene alone increased the density of SERT sites in the mid-frontal cortex and decreased the density of 5-HT(2A)R in the posterior olfactory tubercle. The inhibitory effects of raloxifene on acute estrogen induction of central serotonergic mechanisms were similar to those of tamoxifen even though there are major differences between the two SERMs in their affinity for the two estrogen receptor subtypes and their actions on the uterus. These findings provide robust evidence that estradiol induction of the 5-HT(2A)R and the SERT in brain is mediated by nuclear estrogen receptors. Our data may provide the basis for obtaining a better understanding of the effects of sex steroids on mood and mental state in the human and the possible rational development of congeners of sex steroids for the treatment of mental disorders. PMID- 17398001 TI - Broad spectrum neuroprotection profile of phosphodiesterase inhibitors as related to modulation of cell-cycle elements and caspase-3 activation. AB - Cellular injury can involve the aberrant stimulation of cell cycle proteins in part through activation of phosphodiesterases (PDEs) and downstream expression of cell-cycle components such as cyclin D1. In mature non-proliferating cells activation of the cell cycle can lead to the induction of programmed cell death. In the present study, we investigated the in vitro neuroprotective efficacy and mechanism of action of vinpocetine (PDE1 inhibitor), trequinsin (PDE3 inhibitor), and rolipram (PDE4 inhibitor) in four mechanistically-distinct models of injury to primary rat cortical neurons as related to cell cycle regulation and apoptosis. Cellular injury was induced by hypoxia/hypoglycemia, veratridine (10 microM), staurosporine (1 microM), or glutamate (100 microM), resulting in average neuronal cell death rates of 43-48% as determined by MTT assay. Treatment with each PDE inhibitor (PDEI) resulted in a similar concentration-dependent neuroprotection profile with maximal effective concentrations of 5-10 microM (55 77% neuroprotection) in all four neurotoxicity models. Direct cytotoxicity due to PDE inhibition alone was not observed at concentrations below 100 microM. Further studies indicated that PDEIs can suppress the excitotoxic upregulation of cyclin D1 similar to the effects of flavopiridol, a cyclin-dependent kinase inhibitor, including suppression of pro-apoptotic caspase-3 activity. Overall, these data indicate that PDEIs are broad-spectrum neuroprotective agents acting through modulation of cell cycle elements and may offer a novel mode of therapy against acute injury to the brain. PMID- 17398002 TI - Effects of long-term flutamide treatment during development in zebra finches. AB - The molecular mechanisms responsible for the sexual differentiation of the zebra finch song system remain mysterious. Androgen receptors are expressed in a sexually dimorphic fashion in the zebra finch song system: males have more cells expressing androgen receptors, and this sex difference appears very early in development (day 9 posthatch). Estrogen administration to hatchling females up regulates androgen receptor expression in their song system and profoundly masculinizes their song system's morphology. Co-administering flutamide, an androgen receptor blocker, with estrogen impedes estrogen's masculinizing effects on the song system, suggesting that androgens are required for masculine development. Accordingly, to investigate further the role of androgens in the sexual differentiation of the zebra finch song system, we sought to block androgen activity in males by administering large, sustained doses of flutamide from just before androgen receptors are expressed in the song system (day 7) through to the day of sacrifice (days 61-63). Flutamide profoundly reduced the size of the testes, demonstrating that this drug and mode of administration could have a large impact on tissues. In contrast, flutamide had only a minor impact on the song system: the number of RA neurons was slightly reduced, and the corrected HVC volume showed a trend toward demasculinization. Other brain measures (uncorrected HVC, and corrected and uncorrected volumes of Area X, lMAN, RA, and Rotundus; neuron size in lMAN, HVC, and RA; and number of HVC and LMAN neurons) were not significantly affected. The present results do not support an important role for androgen in masculinizing the song circuit after posthatch day 7. PMID- 17398003 TI - Assessing the role of stimulus probability on the Colavita visual dominance effect. AB - Participants presented with unimodal auditory (A), unimodal visual (V), or bimodal audiovisual stimuli (AV) in a task in which they have to identify the modality of the targets as rapidly as possible, fail to respond to the auditory component of bimodal targets significantly more often than they fail to respond to the visual component. In the majority of published studies on this phenomenon, known as the Colavita effect, the auditory, visual, and bimodal stimuli have been presented in the ratio 40A:40V:20AV. In the present study, we investigated whether the relatively low frequency with which the bimodal targets in previous studies have been presented may have contributed to participants' difficulty in responding to such targets correctly. We manipulated the bimodal target probability by presenting the stimuli in the ratios 20A:20V:60AV, in Experiment 1; 5A:5V:90AV, 25A:25V:50AV, and 45A:45V:10AV, in Experiment 2. A significant Colavita visual dominance effect was observed when the bimodal targets were presented on 60% of the trials or less. We suggest that increasing the frequency of bimodal targets may have provided an exogenous cue to performance, that reduced the necessity for endogenous attention when selecting the appropriate response to make to bimodal targets. PMID- 17398004 TI - NOx and ADMA changes with focal ischemia, amelioration with the chaperonin GroEL. AB - Both nitric oxide and asymmetric dimethylarginine (ADMA) play a critical role in the regulation of cerebral blood flow, though their neuroprotective and cytotoxic effects are still under investigation. In this study, we found that nitrate/nitrite (NOx) levels in plasma, ischemic brain tissue, and cerebrospinal fluid (CSF) increased significantly 24h after 2h transient middle cerebral artery occlusion (MCAO) in rats. ADMA levels were unchanged in plasma, but decreased significantly in CSF 24h following MCAO. The CSF ADMA/NOx ratio decreased markedly following ischemia. Rats protected by expression of the chaperonin GroEL or its folding deficient mutant D87K had lower plasma NOx levels at 24h reperfusion. ADMA, NO, and their ratio in CSF merit further study as biomarkers for ischemic brain injury. PMID- 17398005 TI - Expression and structure of interleukin 4 receptor (IL-4R) complex in human invasive pituitary adenomas. AB - Pituitary adenomas are frequently invasive of surrounding tissues, which adversely affects the surgical outcome and the disease-free survival of patients. In the present study, Interleukin 4 receptor (IL-4R) complex has been investigated to figure out whether the three subunits are overexpressed in human invasive pituitary adenomas. Reverse transcription-polymerase chain reaction (RT PCR) analysis for interleukin 4 receptor alpha (IL-4Ralpha), interleukin 13 receptor alpha1 (IL-13Ralpha1), interleukin 2 receptor gammac (IL-2Rgammac) were performed on total RNA extracted from 10 non-invasive pituitary adenomas, 30 invasive pituitary adenomas, one glioblastoma multiforme, one normal human pituitary tissue sample and one normal human brain tissue sample. Quantitative real-time PCR and in situ immunofluorescence assay were performed in five invasive functioning pituitary adenoma samples and five invasive nonfunctioning pituitary adenoma samples. RT-PCR analysis for IL-4Ralpha, IL-13Ralpha1 and IL 2Rgammac chains were overexpressed in invasive pituitary adenomas. The transcripts for three subunits were not/weakly expressed in normal pituitary tissue and normal brain tissue. The quantitative real-time PCR and in situ immunofluorescence assay confirmed the results of the RT-PCR analysis. Our results indicate that human invasive pituitary adenomas express type III IL-4R complex. These receptors may serve as a novel target for immunotoxin therapy in patients with invasive pituitary adenomas who are not amenable to total surgical resection or for recurrent cases. PMID- 17398007 TI - The contribution of psychosocial factors to the development of chronic pain: the key to better outcomes for patients? PMID- 17398006 TI - A single amino acid in the second transmembrane domain of GABA rho receptors regulates channel conductance. AB - GABAC receptors, expressed predominately in vertebrate retina, are thought to be formed mainly by GABA rho subunits, each of which exhibits distinct physiological and pharmacological properties. In this study, the receptors formed by perch GABA rho subunits were expressed in HEK cells, and their single channel conductances were determined using noise analysis techniques. The receptors formed by the perch rho1A subunit gate a channel with a conductance of 0.2 pS, whereas the receptors formed by GABA rho2 subunits exhibit much higher channel conductances, i.e., 3.2 and 3.5 pS for perch rho2A and rho2B receptors, respectively. A comparison of the amino acid sequences of the channel-forming TMII regions of the various subunits suggested that a single amino acid at position 2' was a potential site for the large differential in conductance. We found that switching the serine residue at that site in the GABA rho2 subunit to the proline residue present in the rho1 subunit reduced the channel conductance to a level similar to that of the wild type rho1 receptor. Conversely, mutating proline to serine in the amino acid sequence of the rho1 receptor significantly increased its unitary conductance. These results indicate that a single amino acid in the TMII region plays an important role in determining the single channel conductance of the GABAC receptors. PMID- 17398008 TI - Performance of local electrodes in the local electrode atom probe. AB - The use of a local electrode in atom probe tomography has enabled higher rates of data acquisition and increased field of view compared to other variants of three dimensional atom probes, but specimen fracture can result in damage to the local electrode. Specimens and local electrodes were examined before and after analyses that resulted in specimen failure. Most specimens were found to be melted after failure and as a result, material was found deposited onto the surface of the local electrode. Material transfer from the specimen to the local electrode was verified by energy dispersive spectrometry in a scanning electron microscope. After the fracture of brittle materials, some remnants were found embedded in the local electrode. For either failure mode, it is likely that the primary specimen rupture produced a sharp protrusion on the specimen or local electrode and this triggered an electrical discharge or uncontrolled field emission that melted a portion of the specimen. The lifetime of the local electrode was found to be dependent on the shape and position of the debris from the specimen failure rather than the number of ions collected or the number of specimens characterized. Local electrodes with smaller apertures were found to be more susceptible to failure. PMID- 17398009 TI - The effects of amount, timing and distribution of simulated rainfall on the development of Haemonchus contortus to the infective larval stage. AB - Three studies were undertaken to determine the effects of amount, timing and distribution of simulated rainfall on the developmental success of Haemonchus contortus. Faeces containing H. contortus eggs were deposited onto pasture plots under a rainfall-activated retractable roof which eliminated incident rainfall. In October (spring) 2004 and January (summer) 2005, the effects of amount (6, 12, 18 or 24 mm) and timing (1, 4, 8 or 15 days post-faecal deposition) of a single simulated rainfall event was investigated via manual application of water to plots. More H. contortus pre-infective larvae (L1 and L2) developed under the d 1 simulated rainfall treatment than later treatments. There was no effect of rainfall amount on development in either experiment, and negligible development to infective larvae (L3). In February (summer) 2006, the effects of amount (12, 24 or 32 mm) and distribution (single event or three smaller but equal split events over 32 h) of simulated rainfall events was investigated with water applied via sprinkler. In this experiment L3 were recovered from the herbage in one-third of the plots harvested, however recovery was low (0.08% of eggs deposited) and there were no treatment effects. Recovery of L1 and L2 from faeces increased with simulated rainfall amount at d 4, and more L1 and L2 were recovered from the split distribution treatment at d 4. The results indicate that moisture conditions soon after faecal deposition are key determinants of H. contortus development success, with significant penalties on development when simulated rainfall was applied 7 days or more post-deposition, and when the duration of simulated rainfall was short. High rates of evaporation during both summer experiments resulted in rapid drying of the micro-environment and this appears to have limited development to L3. PMID- 17398010 TI - A database study that identifies genes whose expression correlates, negatively or positively, with 5-year survival of cancer patients. AB - A published microarray gene expression database containing data on 174 tumor samples from ten tissues was mined, enabling the identification of classes of genes whose expression correlates significantly with the intractability, or tractability, to therapy of tumors derived from such tissues. As a measure of tractability, the 5-year survival of patients presenting with distant (metastatic) tumors was used. Genes that encode proteins related to cell adhesion, and enzymes involved in metabolic oxidation or reduction, were upregulated in intractable cancers. Genes that encode proteins implicated in the control of DNA transcription were downregulated in the intractable cancers. We describe hypotheses with regard to cell functions that may help in designing new therapeutic modalities, aimed at improving survival of cancer patients. PMID- 17398011 TI - Survival from adolescent cancer. AB - BACKGROUND: Adolescent cancer is the leading cause of non-accidental mortality in young people. The tumour types occurring most commonly in this age group are distinctive and differ markedly from those developing in younger children and older adults. OBJECTIVE: The aim is to present survival data for the major types of cancer and for all malignancies combined in adolescents aged from 15 to 19 years, highlighting intercountry differences, temporal trends, and age/treatment regimen comparisons of survival rates. RESULTS: Results from European and American data show that 5-year overall survival among adolescents with cancer is approximately 73-78%, close to that observed among children. As in paediatric series, there are geographical differences, with lower survival rates in Eastern European countries. Adolescents have substantially lower survival than children for acute lymphoid leukaemia, malignant non-Hodgkin lymphoma, Ewing's tumour, osteosarcoma, soft-tissue sarcoma. Temporal improvement in survival from children cancer is twice higher than survival from adolescents malignancies. CONCLUSION: Several reasons could explain these geographical differences, such as better referral, greater availability of complex and expensive treatment regimens, reflecting variations in health care systems and resources. Age-group variations could be explained by differences in biology and in treatment regimen, and lack of participation in clinical trials. Some are inherent in the psychosocial characteristics of these patients, such as delays in seeking medical attention, poor compliance with treatment. It is necessary to focus on cancer in adolescence, as the new frontier in oncology, and adolescents with cancer must be now the subject of specific survival analyses. PMID- 17398012 TI - Predictors of motivation to change alcohol use among referred college students. AB - The current study investigated motivation and its relationship with alcohol use and life satisfaction. Participants were 49 undergraduates referred to a university-based clinic to participate in a brief alcohol intervention. All participants completed measures assessing readiness to change drinking, frequency of alcohol use and related problems, and life satisfaction. As hypothesized, higher levels of motivation to change drinking were related to higher frequency and quantity of alcohol use, alcohol-related problems, and lower levels of life satisfaction. Results indicate the importance of assessing motivation in students referred to receive alcohol interventions. PMID- 17398013 TI - Membranous nephropathy is developed under Th2 environment in chronic graft-versus host disease. AB - Clinical data from case reports of nephrotic syndrome including allogenetic hematopoietic stem cell transplantation (HSCT) suggest that there may be some relationship between chronic graft-versus-host disease (GVHD) and membranous nephropathy (MN). It is widely recognized T cells are crucial for the development of GVHD, and that T helper (Th) cells differentiate into at least two subsets, Th1 and Th2. The polarized situation between Th1 and Th2 cells is established to be important in animal models and human autoimmune diseases. In a chronic GVHD murine model a Th2 cell plays a pivotal role for the pathogenesis. In MRL/lpr mice, which is particularly valuable model for systemic lupus erythematosus, developed diffuse proliferative glomerulonephritis (DPGN) in Th1 environment and MN in Th2 environment. Similarly, Th2 cells may be predominantly activated in chronic GVHD, production and deposition of IgG4 in the glomeruli may develop MN. A hypothesis is: when the patient in period of chronic GVHD developed immune complex-mediated disease, IgG4 might be mainly produced in Th2 environment, and the deposition of IgG4 in the glomeruli may result in the formation of MN. PMID- 17398014 TI - Primary splenic marginal zone lymphoma with florid granulomatous reaction--a case report and review of literature. AB - Splenic marginal zone lymphomas (SMZL) constitute about 20% of primary splenic NHLs. We report a case of primary SMZL with a florid granulomatous reaction which obscured the underlying lymphoma. Although granulomas have been described in splenic non-Hodgkin lymphoma, it can be extensive and mask the underlying lymphoma. A careful search for the cytoarchitectural features of SMZL is warranted in such a case. PMID- 17398015 TI - A model of reoxygenation dynamics of head-and-neck tumors based on serial 18F fluoromisonidazole positron emission tomography investigations. AB - PURPOSE: To develop a model for reoxygenation dynamic and its relationship to local control after radiotherapy (RT), based on repeated dynamic [18F] fluoromisonidazole (FMISO) positron emission tomography (PET) examinations in head-and-neck cancer patients. METHODS AND MATERIALS: Ten head-and-neck cancer patients were examined with dynamic FMISO PET before RT with 70 Gy and after approximately 20 Gy. Two of these patients had two additional dynamic FMISO scans during treatment. Local recurrence was assessed by computed tomography-based follow-up 8-24 months after RT. Tumor-specific values for the level of FMISO retention R and the vascular perfusion efficiency P were determined with a kinetic compartment model. RESULTS: Individual R-P scattergrams before and during therapy were analyzed, and significant therapy-induced changes in the characteristic R-P patterns were observed. A tumor control probability model was derived that involves the tissue parameters R and P and estimates the time to reoxygenation. On the basis of this model, a malignancy value M was introduced and calibrated by a fit to the observed outcome data. Reoxygenation is reflected by the model as a progression to less-malignant tumor types (i.e., smaller values of M). In 4 of 6 patients with severe hypoxia, M had decreased after 20 Gy, whereas 2 patients showed increasing M. Four patients showed no hypoxia in the pretreatment scan. CONCLUSION: A tumor control probability model was developed based on repeated FMISO PET scans during RT. The model combines the local perfusion efficiency and the degree of hypoxia to estimate reoxygenation time. It constitutes a key for hypoxia image-guided dose escalation in RT. PMID- 17398016 TI - Macrophages from irradiated tumors express higher levels of iNOS, arginase-I and COX-2, and promote tumor growth. AB - PURPOSE: To investigate the effects of single and fractionated doses of radiation on tumors and tumor-associated macrophages (TAMs), and to elucidate the potential of TAMs to influence tumor growth. METHODS AND MATERIALS: A murine prostate cell line, TRAMP-C1, was grown in C57Bl/6J mice to 4-mm tumor diameter and irradiated with either 25 Gy in a single dose, or 60 Gy in 15 fractions. The tumors were removed at the indicated times and assessed for a variety of markers related to TAM content, activation status, and function. RESULTS: In tumors receiving a single radiation dose, arginase (Arg-I), and cycloxygenase-2 (COX-2) mRNA expression increased as a small transient wave within 24 h and a larger persistent wave starting after 3 days. Inducible nitric oxide synthase (iNOS) mRNA was elevated only after 3 days and continued to increase up to 3 weeks. After fractionated irradiation, Arg-1 and COX-2 mRNA levels increased within 5 days, whereas iNOS was increased only after 10 fractions of irradiation had been given. Increased levels of Arg-I, COX-2, and, to a lesser extent, iNOS protein were found to associate with TAMs 1-2 weeks after tumor irradiation. Function of TAMs were compared by mixing them with TRAMP-C1 cells and injecting them into mice; TRAMP-C1 cells mixed with TAMs from irradiated tumors appeared earlier and grew significantly faster than those mixed with TAMs from unirradiated tumors or TRAMP-C1 alone. CONCLUSIONS: Tumor-associated macrophages in the postirradiated tumor microenvironment express higher levels of Arg-1, COX-2, and iNOS, and promote early tumor growth in vivo. PMID- 17398017 TI - Can a subgroup of node-negative breast carcinoma patients with T1-2 tumor who may benefit from postmastectomy radiotherapy be identified? AB - PURPOSE: To determine a subgroup of T1-2N0 breast carcinoma patients at high risk for local recurrence. METHODS AND MATERIALS: In this retrospective study, univariate and multivariate prognostic factor analyses for local recurrence and distant recurrence were carried out in 502 patients. RESULTS: During the median observation time of 77 months (range, 24-191 months), 14 patients (2.8%) had local recurrence and 55 (11.0%) had distant recurrence. Tumor size (continuous, p = 0.03; hazard ratio [HR] 1.2; 95% confidence interval [CI], 1.1-1.7), grade (p = 0.01; HR, 2.4; 95% CI, 1.2-5.0), lymphatic vascular invasion (LVI) (p = 0.01; HR, 10.0; 95% CI, 2.4-17.3), estrogen receptor status (p = 0.01; HR, 6.3; 95% CI, 2.0 23.0) and cErbB2 status (p = 0.01; HR, 10.0; 95% CI 1.8-87.5) were strongly associated with distant recurrence. Tumor size (< or =2 cm vs. >2 cm; p = 0.05; HR, 5.4; 95% CI, 1.2-28.0) and LVI (p = 0.004; HR, 9.0; 95% CI, 2.0-41.0) in patients aged < or =40 years, and tumor size (< or =3 cm vs. >3 cm; p = 0.05; HR 8.6; 95% CI 1.3-75.0), LVI (p = 0.007; HR, 18.0; 95% CI, 2.1-153.0), and grade (p = 0.05; HR, 7.0; 95% CI, 1.2-63.0) in patients aged >40 years were the most important predictive factors for local recurrence. CONCLUSIONS: Among breast carcinoma patients, young patients with tumor size >2 cm and LVI and older patients with tumor size >3 cm, LVI, and high-grade tumor had a high risk of local recurrence. PMID- 17398018 TI - PET-CT-based auto-contouring in non-small-cell lung cancer correlates with pathology and reduces interobserver variability in the delineation of the primary tumor and involved nodal volumes. AB - PURPOSE: To compare source-to-background ratio (SBR)-based PET-CT auto delineation with pathology in non-small-cell lung cancer (NSCLC) and to investigate whether auto-delineation reduces the interobserver variability compared with manual PET-CT-based gross tumor volume (GTV) delineation. METHODS AND MATERIALS: Source-to-background ratio-based auto-delineation was compared with macroscopic tumor dimensions to assess its validity in 23 tumors. Thereafter, GTVs were delineated manually on 33 PET-CT scans by five observers for the primary tumor (GTV-1) and the involved lymph nodes (GTV-2). The delineation was repeated after 6 months with the auto-contour provided. This contour was edited by the observers. For comparison, the concordance index (CI) was calculated, defined as the ratio of intersection and the union of two volumes (A intersection B)/(A union or logical sum B). RESULTS: The maximal tumor diameter of the SBR-based auto-contour correlated strongly with the macroscopic diameter of primary tumors (correlation coefficient = 0.90) and was shown to be accurate for involved lymph nodes (sensitivity 67%, specificity 95%). The median auto-contour-based target volumes were smaller than those defined by manual delineation for GTV-1 (31.8 and 34.6 cm(3), respectively; p = 0.001) and GTV-2 (16.3 and 21.8 cm(3), respectively; p = 0.02). The auto-contour-based method showed higher CIs than the manual method for GTV-1 (0.74 and 0.70 cm(3), respectively; p < 0.001) and GTV-2 (0.60 and 0.51 cm(3), respectively; p = 0.11). CONCLUSION: Source-to-background ratio-based auto-delineation showed a good correlation with pathology, decreased the delineated volumes of the GTVs, and reduced the interobserver variability. Auto-contouring may further improve the quality of target delineation in NSCLC patients. PMID- 17398019 TI - Prognostic factors depicting disease-specific survival in parotid-gland tumors. AB - PURPOSE: To identify significant prognostic factors that can be used in clinical decision-making with regard to parotid cancer, which is characterized by a complex and diverse group of tumors with variable outcomes. METHODS AND MATERIALS: A historical cohort of 184 patients with parotid-gland malignancy, who had been registered in the Province of Manitoba from 1970 to 2003, was examined. Survival analysis was performed using Kaplan-Meier curves and a log-rank test for comparing subgroups. The independent effect of factors that predicted survival at the bivariate level was determined using a Cox proportional hazard model. RESULTS: The mean age at presentation was 62 years. The mean follow-up was 64 months. Absolute and disease-specific survival at 5 years was 41.70% and 57.94%, respectively. Survival for Stages I-IV at 5 years was 85.35%, 76.9%, 56.1%, and 8.4%, respectively (p < 0.0001). Factors with an independent effect on survival (p < 0.05) included age, tumor size, local invasion (Stages T4 vs. T1), and distant metastasis at presentation, tumor differentiation, and treatment. Adjuvant radiotherapy vs. surgery alone reduced the risk of death from disease at 5 years by 50% (hazard ratio [HR], 0.5; 95% confidence interval, 0.228-0.995; p = 0.0486). CONCLUSIONS: Despite the variety of malignant parotid tumors, easily identifiable prognostic indicators such as advanced age, tumor size, local invasion, and tumor differentiation have a significant impact on outcome. Patients with adverse prognostic factors benefit from adjuvant radiotherapy. The threshold for the use of adjuvant radiotherapy in managing parotid malignancy should be low. PMID- 17398020 TI - Fatigue during head-and-neck radiotherapy: prospective study on 117 consecutive patients. AB - PURPOSE: Fatigue is an underevaluated cancer-related and treatment-related symptom. We analyzed fatigue in head and neck cancer patients undergoing radiotherapy (RT). METHODS AND MATERIALS: A total of 117 patients were enrolled (mean age, 58 years). Radiation therapy (median dose, 66 Gy) was given with either exclusive or postoperative intent in 52 and 65 patients, respectively. Chemotherapy (CT) was added before and/or during RT in 61 patients. The patients completed a 20-item questionnaire (Multidimensional Fatigue Inventory [MFI-20]) before, during (weekly), and after RT. The impact of patient-, tumor-, and treatment-related factors on fatigue was evaluated with unifactorial and multifactorial tests. RESULTS: Fatigue level increased during RT reaching a maximum at Week 6 and then slowly decreased. In multivariate stepwise regression analysis age (inversely related, p < 0.05), psychologic disorders (p < 0.005), and previous head-and-neck surgery (inversely related, p < 0.005) were correlated with higher pre-RT fatigue level. Pre-RT fatigue score (p < 0.0001), induction and/or concomitant CT (p = 0.035), need of cortisone during RT (p = 0.005), and thyroid disorders (p = 0.032) were correlated with higher during-RT fatigue level. Pre-RT fatigue score (p < 0.0001), induction and/or concomitant CT (p < 0.001), and need of cortisone during RT (p < 0.005) were correlated with higher post-RT fatigue level. No impact of gender, performance status, comorbidities other than psychologic and thyroid, tumor stage/site, RT intent, dose, volume, duration, or toxicity was observed. CONCLUSION: Fatigue affects all patients undergoing RT for head-and-neck cancer, reaches maximum score at the 6th week of RT, and slowly decreases thereafter. Age, thyroid dysfunction, psychologic disorders, pre-RT fatigue score, CT, and cortisone use are correlated with RT related fatigue levels. PMID- 17398021 TI - Kilo-voltage cone-beam computed tomography setup measurements for lung cancer patients; first clinical results and comparison with electronic portal-imaging device. AB - PURPOSE: Kilovoltage cone-beam computed tomography (CBCT) has been developed to provide accurate soft-tissue and bony setup information. We evaluated clinical CBCT setup data and compared CBCT measurements with electronic portal imaging device (EPID) images for lung cancer patients. METHODS AND MATERIALS: The setup error for CBCT scans at the treatment unit relative to the planning CT was measured for 62 patients (524 scans). For 19 of these patients (172 scans) portal images were also made. The mean, systematic setup error (Sigma), and random setup error (sigma) were calculated for the CBCT and the EPID. The differences between CBCT and EPID and the rotational setup error derived from the CBCT were also evaluated. An offline shrinking action level correction protocol, based on the CBCT measurements, was used to reduce systematic setup errors and the impact of this protocol was evaluated. RESULTS: The CBCT setup errors were significantly larger than the EPID setup errors for the cranial-caudal and anterior-posterior directions (p < 0.05). The mean overall setup errors after correction measured with the CBCT were 0.2 mm (Sigma = 1.6 mm, sigma = 2.9 mm) in the left-right, 0.8 mm (Sigma = 1.7 mm, sigma = 4.0 mm) in cranial-caudal and 0.0 mm (Sigma = 1.5 mm, sigma = 2.0 mm) in the anterior-posterior direction. Using our correction protocol only 2 patients had mean setup errors larger than 5 mm, without this correction protocol 51% of the patients would have had a setup error larger than 5 mm. CONCLUSION: Use of CBCT scans provided more accurate information concerning the setup of lung cancer patients than did portal imaging. PMID- 17398022 TI - Risk, outcomes, and costs of radiation-induced oral mucositis among patients with head-and-neck malignancies. AB - PURPOSE: To study the risk, outcomes, and costs of radiation-induced oral mucositis (OM) among patients receiving radiotherapy (RT) to head and neck primary cancers. METHODS AND MATERIALS: A retrospective cohort consisting of 204 consecutive head-and-neck cancer patients who received RT with or without chemotherapy during 2002 was formed; their records were reviewed for clinical and resource use information. Patients who had received prior therapy, had second primary cancers, or received palliative radiation therapy were excluded. The risk of OM was analyzed by multiple variable logistic regression. The cost of care was computed from the provider's perspective in 2006 U.S. dollars and compared among patients with and without OM. RESULTS: Oral mucositis occurred in 91% of patients; in 66% it was severe (Grade 3-4). Oral mucositis was more common among patients with oral cavity or oropharynx primaries (odds ratio [OR], 44.5; 95% confidence interval [CI], 5.2 to >100; p < 0.001), those who received chemotherapy (OR = 7.8; 95% CI, 1.5-41.6; p = 0.02), and those who were treated with altered fractionation schedules (OR = 6.3; 95% CI, 1.1-35.1; p = 0.03). Patients with OM were significantly more likely to have severe pain (54% vs. 6%; p < 0.001) and a weight loss of > or =5% (60% vs. 17%; p < 0.001). Oral mucositis was associated with an incremental cost of $1700-$6000, depending on the grade. CONCLUSIONS: Head-and-neck RT causes OM in virtually all patients. Oral mucositis is associated with severe pain, significant weight loss, increased resource use, and excess cost. Preventive strategies are needed. PMID- 17398023 TI - Analysis of biochemical control and prognostic factors in patients treated with either low-dose three-dimensional conformal radiation therapy or high-dose intensity-modulated radiotherapy for localized prostate cancer. AB - PURPOSE: To identify prognostic factors and evaluate biochemical control rates for patients with localized prostate cancer treated with either high-dose intensity-modulated radiotherapy (IMRT) or conventional-dose three-dimensional conformal radiotherapy 3D-CRT. METHODS: Four hundred sixteen patients with a minimum follow-up of 3 years (median, 5 years) were included. Two hundred seventy one patients received 3D-CRT with a median dose of 68.4 Gy (range, 66-71 Gy). The next 145 patients received IMRT with a median dose of 75.6 Gy (range, 70.2-77.4 Gy). Biochemical control rates were calculated according to both American Society for Therapeutic Radiology and Oncology (ASTRO) consensus definitions. Prognostic factors were identified using both univariate and multivariate analyses. RESULTS: The 5-year biochemical control rate was 60.4% for 3D-CRT and 74.1% for IMRT (p < 0.0001, first ASTRO Consensus definition). Using the ASTRO Phoenix definition, the 5-year biochemical control rate was 74.4% and 84.6% with 3D-RT and IMRT, respectively (p = 0.0326). Univariate analyses determined that PSA level, T stage, Gleason score, perineural invasion, and radiation dose were predictive of biochemical control. On multivariate analysis, dose, Gleason score, and perineural invasion remained significant. CONCLUSION: On the basis of both ASTRO definitions, dose, Gleason score, and perineural invasion were predictive of biochemical control. Intensity-modulated radiotherapy allowed delivery of higher doses of radiation with very low toxicity, resulting in improved biochemical control. PMID- 17398024 TI - Dosimetric predictors of laryngeal edema. AB - PURPOSE: To investigate dosimetric predictors of laryngeal edema after radiotherapy (RT). METHODS AND MATERIALS: A total of 66 patients were selected who had squamous cell carcinoma of the head and neck with grossly uninvolved larynx at the time of RT, no prior major surgical operation except for neck dissection and tonsillectomy, treatment planning data available for analysis, and at least one fiberoptic examination of the larynx within 2 years from RT performed by a single observer. Both the biologically equivalent mean dose at 2 Gy per fraction and the cumulative biologic dose-volume histogram of the larynx were extracted for each patient. Laryngeal edema was prospectively scored after treatment. Time to endpoint, moderate or worse laryngeal edema (Radiation Therapy Oncology Group Grade 2+), was calculated with log rank test from the date of treatment end. RESULTS: At a median follow-up of 17.1 months (range, 0.4- 50.0 months), the risk of Grade 2+ edema was 58.9% +/- 7%. Mean dose to the larynx, V30, V40, V50, V60, and V70 were significantly correlated with Grade 2+ edema at univariate analysis. At multivariate analysis, mean laryngeal dose (continuum, hazard ratio, 1.11; 95% confidence interval, 1.06-1.15; p < 0.001), and positive neck stage at RT (N0-x vs. N +, hazard ratio, 3.66; 95% confidence interval, 1.40 9.58; p = 0.008) were the only independent predictors. Further stratification showed that, to minimize the risk of Grade 2+ edema, the mean dose to the larynx has to be kept < or =43.5 Gy at 2 Gy per fraction. CONCLUSION: Laryngeal edema is strictly correlated with various dosimetric parameters; mean dose to the larynx should be kept < or =43.5 Gy. PMID- 17398025 TI - Spatial and dosimetric variability of organs at risk in head-and-neck intensity modulated radiotherapy. AB - PURPOSE: The accuracy of intensity-modulated radiotherapy (IMRT) delivery may be compromised by random spatial error and systematic anatomic changes during the treatment course. We present quantitative measurements of the spatial variability of head-and-neck organs-at-risk and demonstrate the resultant dosimetric effects. METHODS AND MATERIALS: Fifteen consecutive patients were imaged weekly using computed tomography during the treatment course. Three-dimensional displacements were calculated for the superior and inferior brainstem; C1, C6, and T2 spinal cord; as well as the lateral and medial aspects of the parotid glands. The data were analyzed to show distributions of spatial error and to track temporal changes. The treatment plan was recalculated on all computed tomography sets, and the dosimetric error was quantified in terms of the maximal dose difference (brainstem and spinal cord) or the mean dose difference and the volume receiving 26 Gy (parotid glands). RESULTS: The mean three-dimensional displacement was 2.9 mm for the superior brainstem, 3.4 mm for the inferior brainstem, 3.5 mm for the C1 spine, 5.6 mm for the C6 spine and 6.0 mm for the T2 spine. The lateral aspects of both parotid glands showed a medial translation of 0.85 mm/wk, and glands shrank by 4.9%/wk. The variability of the maximal dose difference was described by standard deviations ranging from 5.6% (upper cord) to 8.0% (lower cord.) The translation of the left parotid resulted in an increase of the mean dose and the volume receiving 26 Gy. CONCLUSION: Random spatial and dosimetric variability is predominant for the brainstem and spinal cord and increases at more inferior locations. In contrast, the parotid glands demonstrated a systematic medial translation during the treatment course and thus sparing may be compromised. PMID- 17398026 TI - What dose of external-beam radiation is high enough for prostate cancer? AB - PURPOSE: To quantify the radiotherapy dose-response of prostate cancer, adjusted for prognostic factors in a mature cohort of men treated relatively uniformly at a single institution. PATIENTS AND METHODS: The study cohort consisted of 1,530 men treated with three-dimensional conformal external-beam radiotherapy between 1989 and 2002. Patients were divided into four isocenter dose groups: <70 Gy (n = 43), 70-74.9 Gy (n = 552), 75-79.9 Gy (n = 568), and > or =80 Gy (n = 367). The primary endpoints were freedom from biochemical failure (FFBF), defined by American Society for Therapeutic Radiology and Oncology (ASTRO) and Phoenix (nadir + 2.0 ng/mL) criteria, and freedom from distant metastases (FFDM). Multivariate analyses were performed and adjusted Kaplan-Meier estimates were calculated. Logit regression dose-response functions were determined at 5 and 8 years for FFBF and at 5 and 10 years for FFDM. RESULTS: Radiotherapy dose was significant in multivariate analyses for FFBF (ASTRO and Phoenix) and FFDM. Adjusted 5-year estimates of ASTRO FFBF for the four dose groups were 60%, 68%, 76%, and 84%. Adjusted 5-year Phoenix FFBFs for the four dose groups were 70%, 81%, 83%, and 89%. Adjusted 5-year and 10-year estimates of FFDM for the four dose groups were 96% and 93%, 97% and 93%, 99% and 95%, and 98% and 96%. Dose response functions showed an increasing benefit for doses > or =80 Gy. CONCLUSIONS: Doses of > or =80 Gy are recommended for most men with prostate cancer. The ASTRO definition of biochemical failure does not accurately estimate the effects of radiotherapy at 5 years because of backdating, compared to the Phoenix definition, which is less sensitive to follow-up and more reproducible over time. PMID- 17398027 TI - Proton-beam therapy for olfactory neuroblastoma. AB - PURPOSE: To analyze the feasibility and efficacy of proton-beam therapy (PBT) for olfactory neuroblastoma (ONB) as a definitive treatment, by reviewing our preliminary experience. Olfactory neuroblastoma is a rare disease, and a standard treatment strategy has not been established. Radiation therapy for ONB is challenging because of the proximity of ONBs to critical organs. Proton-beam therapy can provide better dose distribution compared with X-ray irradiation because of its physical characteristics, and is deemed to be a feasible treatment modality. METHODS AND MATERIALS: A retrospective review was performed on 14 patients who underwent PBT for ONB as definitive treatment at the National Cancer Center Hospital East (Kashiwa, Chiba, Japan) from November 1999 to February 2005. A total dose of PBT was 65 cobalt Gray equivalents (Gy(E)), with 2.5-Gy(E) once daily fractionations. RESULTS: The median follow-up period for surviving patients was 40 months. One patient died from disseminated disease. There were two persistent diseases, one of which was successfully salvaged with surgery. The 5 year overall survival rate was 93%, the 5-year local progression-free survival rate was 84%, and the 5-year relapse-free survival rate was 71%. Liquorrhea was observed in one patient with Kadish's stage C disease (widely destroying the skull base). Most patients experienced Grade 1 to 2 dermatitis in the acute phase. No other adverse events of Grade 3 or greater were observed according to the RTOG/EORTC acute and late morbidity scoring system. CONCLUSIONS: Our preliminary results of PBT for ONB achieved excellent local control and survival outcomes without serious adverse effects. Proton-beam therapy is considered a safe and effective modality that warrants further study. PMID- 17398029 TI - Risk of hemorrhage and obliteration rates of LINAC-based radiosurgery for cerebral arteriovenous malformations treated after prior partial embolization. AB - PURPOSE: We investigate patient outcome, risk of hemorrhage, and factors affecting obliteration after LINAC-based radiosurgery (RS) for cerebral arteriovenous malformations (AVM) treated after partial embolization. METHODS AND MATERIALS: This analysis is based on 50 patients treated after prior embolization. According to the Spetzler-Martin criteria the AVM classification was as follows: 9 patients, Grade I (18%); 19, Grade II (38%); 18, Grade III (36%); and 4, Grade IV (8%). Median RS-based AVM score was 1.41. Median single dose was 18 Gy/ 80% isodose (range, 15-22 Gy) and median AVM volume was 4.0 cc (range, 0.2-22.6 cc). In all, 34 patients (68%) experienced hemorrhage before RS. Median follow-up was 3.1 year (range, 8.5 months to 15 years). RESULTS: Actuarial complete obliteration rate was 67% after 3 years and 78% after 4 years. The complete obliteration rate was significantly higher in AVM <3 cm (92% vs. 60%, p < 0.01) and in AVM Spetzler-Martin Grade I/II (90% vs. 59%, p < 0,01). Intracranial hemorrhage after RS was seen in 6 patients (12%) after 8.5 months median. Annual bleeding risk was 7.9% after 1 year and 2.2% after 2 years. It was found that AVM diameter > or =3 cm (p < 0.006), AVM volume > or =4 cc (p < 0.01), AVM score > or =1.5 (p < 0.03), and single dose <18 Gy (p < 0.03) were associated with a significant higher bleeding risk. CONCLUSIONS: The rate of obliteration after RS in AVM treated after prior partial embolization depends on size as well as Spetzler-Martin grade. The risk of intracranial hemorrhage is not increased after RS and depends on AVM score, size, and volume, as well as on applied single dose. PMID- 17398028 TI - Reduction of normal lung irradiation in locally advanced non-small-cell lung cancer patients, using ventilation images for functional avoidance. AB - PURPOSE: To investigate the ability of four-dimensional computed tomography (4D CT)-derived ventilation images to identify regions of highly functional lung for avoidance in intensity-modulated radiotherapy (IMRT) planning in locally advanced non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: The treatment-planning records from 21 patients with Stage III NSCLC were selected. Ventilation images were generated from the 4D-CT sets, and each was imported into the treatment planning system. Ninetieth percentile functional volumes (PFV90), constituting the 10% of the lung volume where the highest ventilation occurs, were generated. Baseline IMRT plans were generated using the lung volume constraint on V20 (<35%), and two additional plans were generated using constraints on the PFV90 without a volume constraint. Dose-volume and dose-function histograms (DVH, DFH) were generated and used to evaluate the planning target volume coverage, lung volume, and functional parameters for comparison of the plans. RESULTS: The mean dose to the PFV90 was reduced by 2.9 Gy, and the DFH at 5 Gy (F5) was reduced by 9.6% (SE = 2.03%). The F5, F10, V5, and V10 were all significantly reduced from the baseline values. We identified a favorable subset of patients for whom there was a further significant improvement in the mean lung dose. CONCLUSIONS: Four dimensional computed tomography-derived ventilation regions were successfully used as avoidance structures to reduce the DVH and DFH at 5 Gy in all cases. In a subset, there was also a reduction in the F10 and V10 without a change in the V20, suggesting that this technique could be safely used. PMID- 17398030 TI - Selective mastectomy in the management of locally advanced breast cancer. AB - PURPOSE: To evaluate local control for patients with locally advanced noninflammatory breast cancer (LABC) managed by selective mastectomy. METHODS AND MATERIALS: Between 1979 and 1996, 176 patients with LABC were prospectively managed by chemotherapy (CT)-irradiation (RT)-CT without routine mastectomy. All surviving patients were followed for a minimum of 5 years. RESULTS: A total of 132 patients (75%) had a T4 tumor and 22 (12.5%) supraclavicular nodal disease. The clinical complete response rate was 91% (160/176), which included 13 patients who underwent mastectomy and 2 an iridium wire implant. The first site of failure was local for 43 patients (breast +/- axilla for 38); 27 of these patients underwent salvage mastectomy and 11 did not for an overall mastectomy rate of 23% (40/176). If all 176 patients had undergone routine mastectomy (136 extra mastectomies), 11 additional patients may have avoided an unsalvageable first local relapse. The others would have either have not had a local relapse or would have suffered local relapse after distant disease. No tumor or treatment related factor was found to predict local disease at death. Median disease-free and overall survival for all patients was 26 and 52 months, respectively. CONCLUSIONS: Selective mastectomy in LABC may not jeopardize local control or survival. PMID- 17398031 TI - Extended-field irradiation and intracavitary brachytherapy combined with cisplatin chemotherapy for cervical cancer with positive para-aortic or high common iliac lymph nodes: results of ARM 1 of RTOG 0116. AB - PURPOSE: The Radiation Therapy Oncolology Group (RTOG) 0116 trial was designed to test the ability of Amifostine to reduce the toxicity of combined chemotherapy with extended-field radiotherapy and brachytherapy (Part 2), after first determining the toxicity rate for the regimen without Amifostine (Part 1). This manuscript reports the results of Part 1. METHODS AND MATERIALS: Eligibility included patients with cervical carcinoma and high common iliac or para-aortic metastasis. Patients received extended-field radiotherapy to 45 Gy (1.8 Gy/fraction) with intracavitary irradiation. The final point A dose was 85 Gy LDR equivalent. Use of HDR was allowed. The positive para-aortic and high common iliac nodes were boosted to 54 to 59.4 Gy. Cisplatin (40 mg/m(2)) was delivered weekly during external beam and once with brachytherapy. The primary endpoint of Part 1 was acute Grade 3/4 toxicity, excluding Grade 3 leukopenia. RESULTS: A total of 26 eligible patients were entered between August 1, 2000, and December 3, 2003. Of these, 21 had para-aortic metastasis (15 also had high common iliac involvement), and 5 had high common iliac involvement only. The median follow-up was 17.1 months (range, 1.8-38.6 months) for all patients and 21.7 months (range, 11.4-38.6 months) for alive patients. The acute Grade 3/4 toxicity rate, excluding Grade 3 leukopenia was 81%. Late Grade 3/4 toxicity was 40%. Eight patients underwent surgery for complications. Sixteen (62%) patients had a complete response for both local and nodal disease. The complete local response was 92%, the complete overall nodal response rate was 62% and the regional and para-aortic nodal response rates were 60% and 71% respectively. Estimated disease free and overall survival at 18 months are 46% and 60%. CONCLUSIONS: Extended field and intracavitary irradiation with cisplatin for para-aortic or high common iliac metastasis from cervical cancer is associated with significant acute and late toxicity. PMID- 17398032 TI - Perineural invasion is a marker for pathologically advanced disease in localized prostate cancer. AB - PURPOSE: To determine if perineural invasion (PNI) should be included in addition to prostate-specific antigen (PSA), biopsy Gleason score, and clinical T-stage for risk-stratification of patients with localized prostate cancer. METHODS AND MATERIALS: We analyzed prostatectomy findings for 1550 patients, from a prospectively collected institutional database, to determine whether PNI was a significant predictor for upgrading of Gleason score or pathologic T3 disease after patients were stratified into low-, intermediate-, and high-risk groups (on the basis of PSA, biopsy Gleason score, and clinical T-stage). RESULTS: For the overall population, PNI was associated with a significantly increased frequency of upgrading and of pathologic T3 disease. After stratification, PNI was still associated with significantly increased odds of pathologic T3 disease within each risk group. In particular, for low-risk patients, there was a markedly increased risk of extraprostatic extension (23% vs. 7%), comparable to that of intermediate risk patients. Among high-risk patients, PNI was associated with an increased risk of seminal vesicle invasion and lymph node involvement. Furthermore, over 80% of high-risk patients with PNI were noted to have an indication for postoperative radiation. CONCLUSIONS: Perineural invasion may be useful for risk stratification of prostate cancer. Our data suggest that low-risk patients with PNI on biopsy may benefit from treatment typically reserved for those with intermediate-risk disease. In addition, men with high-risk disease and PNI, who are contemplating surgery, should be informed of the high likelihood of having an indication for postoperative radiation therapy. PMID- 17398033 TI - Radiation pneumonitis: local dose versus [18F]-fluorodeoxyglucose uptake response in irradiated lung. AB - PURPOSE: To quantify the relationship between the local radiation dose received and the posttreatment positron emission tomography/computed tomography (PET/CT) [(18)F]2-fluoro-2-deoxyglucose (FDG) uptake in the lung. METHODS AND MATERIALS: The data from 36 patients treated for esophageal cancer with thoracic radiotherapy who underwent restaging PET/CT imaging between 4 and 12 weeks after radiotherapy completion were evaluated. Their treatment planning CT was registered with the restaging PET/CT. Using histogram analysis, the voxel average FDG-PET uptake vs. radiation dose was obtained for each case. Hierarchical linear regression models for each patient were applied to study the variation in the linear trends between cases. Deviation of the dose-response curve from a linear model was tested. RESULTS: The median time between radiotherapy completion and FDG-PET imaging was 40 days (range, 26-70 days). The median of the mean standard uptake value in the lung that received 0-5 Gy was 0.63 (range, 0.36-1.27), 5-10 Gy was 0.77 (range, 0.40-1.35), 10-20 Gy was 0.80 (range, 0.40-1.72), and >20 Gy was 1.08 (range, 0.44-2.63). A hierarchical linear regression model of the radiation dose and normalized FDG uptake per case found an adequate fit with the linear model, and the addition of quadratic and logarithmic functions did not improve the fit. The 36 cases had a posterior mean of slopes range of 0.0048 0.069. CONCLUSION: The regional dose vs. radiation pneumonitis response was evaluated with FDG-PET/CT imaging. Statistical modeling found a linear relationship. The slope of this relationship varied over an order of magnitude, reflecting the range of the underlying biological response to radiation among the study population. PMID- 17398034 TI - Cyclin D1 gene G870A polymorphism predicts response to neoadjuvant radiotherapy and prognosis in rectal cancer. AB - PURPOSE: To investigate whether CCND1 genetic variations associated with a constitutive nuclear protein may influence either the pathologic response to preoperative RT or the prognosis in a series of rectal cancer patients. METHODS AND MATERIALS: Seventy rectal cancer patients treated by neoadjuvant radiotherapy were included in the study. CCND1 exon 5 mutations were screened, and the G870A polymorphism was assessed for correlation with clinical variables, tumor response, and patient outcome. RESULTS: No exon 5 mutation was found. Concerning the G870A polymorphism, the A/A variant was significantly associated with radiosensitivity (p = 0.022). Moreover, patients harboring the A allele were correlated with a lower risk of local failure (p = 0.017). Also, combination of the G870A polymorphism with the post-therapeutic lymph node status allowed the elaboration of a prognostic index, which accurately distinguished subgroups of patients with predictable recurrence-free (p = 0.003) and overall (p = 0.044) survival. CONCLUSIONS: Although CCND1 exon 5 mutations are rare in rectal cancer, G870A polymorphism is a frequent variation that may predict radiosensitivity and prognosis. PMID- 17398035 TI - Assessing respiration-induced tumor motion and internal target volume using four dimensional computed tomography for radiotherapy of lung cancer. AB - PURPOSE: To assess three-dimensional tumor motion caused by respiration and internal target volume (ITV) for radiotherapy of lung cancer. METHODS AND MATERIALS: Respiration-induced tumor motion was analyzed for 166 tumors from 152 lung cancer patients, 57.2% of whom had Stage III or IV non-small-cell lung cancer. All patients underwent four-dimensional computed tomography (4DCT) during normal breathing before treatment. The expiratory phase of 4DCT images was used as the reference set to delineate gross tumor volume (GTV). Gross tumor volumes on other respiratory phases and resulting ITVs were determined using rigid-body registration of 4DCT images. The association of GTV motion with various clinical and anatomic factors was analyzed statistically. RESULTS: The proportions of tumors that moved >0.5 cm along the superior-inferior (SI), lateral, and anterior posterior (AP) axes during normal breathing were 39.2%, 1.8%, and 5.4%, respectively. For 95% of the tumors, the magnitude of motion was less than 1.34 cm, 0.40 cm, and 0.59 cm along the SI, lateral, and AP directions. The principal component of tumor motion was in the SI direction, with only 10.8% of tumors moving >1.0 cm. The tumor motion was found to be associated with diaphragm motion, the SI tumor location in the lung, size of the GTV, and disease T stage. CONCLUSIONS: Lung tumor motion is primarily driven by diaphragm motion. The motion of locally advanced lung tumors is unlikely to exceed 1.0 cm during quiet normal breathing except for small lesions located in the lower half of the lung. PMID- 17398036 TI - Neural stem cells: implications for the conventional radiotherapy of central nervous system malignancies. AB - Advances in basic neuroscience related to neural stem cells and their malignant counterparts are challenging traditional models of central nervous system tumorigenesis and intrinsic brain repair. Neurogenesis persists into adulthood predominantly in two neurogenic centers: subventricular zone and subgranular zone. Subventricular zone is situated adjacent to lateral ventricles and subgranular zone is confined to the dentate gyrus of the hippocampus. Neural stem cells not only self-renew and differentiate along multiple lineages in these regions, but also contribute to intrinsic brain plasticity and repair. Ionizing radiation can depopulate these exquisitely sensitive regions directly or impair in situ neurogenesis by indirect, dose-dependent and inflammation-mediated mechanisms, even at doses <2 Gy. This review discusses the fundamental neural stem cell concepts within the framework of cumulative clinical experience with the treatment of central nervous system malignancies using conventional radiotherapy. PMID- 17398037 TI - [Postpartum compliance to oral glucose tolerance test]. AB - OBJECTIVES: The main objective of this study is to compare the rate of adherence to GTT done six weeks postpartum (early GTT) to the one performed three months or more after delivery (late GTT). The secondary objective is to determine factors associated to non-adherence to GTT. MATERIALS AND METHODS: This is a retrospective study involving women with GDM who delivered in 1998 (late GTT; N=229) and 2003 (early GTT; N=244). We retrieved maternal, pregnancy, newborn and GDM parameters. Statistical analyses include the chi2 test and logistic regression analysis. RESULTS: The rate of adherence to early GTT (77.1%) was statistically higher than the one observed for late GTT (45.6%; P<0.001). The factors identified as predictors of non-adherence were the year of delivery, parity > or = 2, problem of adherence to GDM treatment, and no breastfeeding six weeks after delivery. CONCLUSION: GTT performed six weeks after delivery increases the adherence to the test. The factors associated with non-adherence are more related to personal matters than to GDM. PMID- 17398038 TI - Haloperidol, dynamics of choice, and the parameters of the matching law. AB - The idea that dopamine mediates the reinforcing effects of stimuli persists in the field of neurosciences. The present study shows that haloperidol, a dopamine antagonist, does not eliminate the reinforcing value of food reinforcers. The ratio of reinforcers changed seven times across two levers within sessions, modeling a dynamic environment. The magnitude of the reinforcer was manipulated independently of the reinforcer ratio. Four doses of intraperitoneal haloperidol were assessed over periods of 12 daily sessions. Haloperidol did not impair the discrimination that the rats established between rich and lean levers; the response distributions favored the lever associated with the higher probability of reinforcement and the larger pellets. The parameters of the generalized matching law (bias and sensitivity) were used to estimate effects of haloperidol upon the motor system and upon the rats' motivation for food reinforcers. PMID- 17398039 TI - Distributive syringe sharing among young adult injection drug users in five U.S. cities. AB - Blood-borne pathogens such as human immunodeficiency virus (HIV) and hepatitis C virus (HCV) are transmitted most commonly among injection drug users (IDUs) through the sharing of needles and syringes. Distributive syringe sharing (DSS) (i.e., passing on a used needle/syringe to another IDU) poses the potential risk of transmitting HIV and viral hepatitis to others. We studied the prevalence and correlates of DSS among IDUs enrolled in a randomized behavioral intervention trial designed to reduce behaviors associated with HIV and HCV transmission in five U.S. cities. Among 3129 IDUs ages 15-30 years who completed the baseline visit, 1432 (45.8%) engaged in DSS during the 3 months prior to baseline. Significant correlates of DSS were perception that peer norms condone needle sharing, frequent injection, not obtaining most syringes from needle exchange programs or pharmacies, injecting most frequently in shooting galleries and with sex partners, low perceived risk of HIV from sharing syringes, increased anxiety, low self-esteem, and having unprotected sex. Restricting to only those IDUs who reported not injecting with previously used syringes, similar independent correlates of DSS were found. These findings suggest that interventions to reduce ongoing transmission of blood-borne infections should focus on altering peer norms among networks of young IDUs. PMID- 17398040 TI - Initiation into methamphetamine use for young gay and bisexual men. AB - Research over the past 10 years has suggested that methamphetamine use has become a significant problem and is associated with risky sexual behaviors among gay and bisexual men. In order to better understand initiation into methamphetamine use among gay and bisexual men, qualitative analyses were performed on a sample of young gay and bisexual men (ages 18-29) in New York City. Participants were recruited as part of a larger study which used time-space sampling to enroll club going young adults who indicated recent club drug (ecstasy, ketamine, GHB, methamphetamine, cocaine, and/or LSD) use. The data for this paper are derived from the qualitative interviews of 54 gay and bisexual male methamphetamine users. At initiation (1) methamphetamine was used in a social, non-sexual setting for a majority of the participants; (2) participants expressed limited knowledge of methamphetamine; and (3) many participants used cocaine as a basis for comparison when describing various effects of the drug. The understanding that at initiation methamphetamine was not solely used as a sexual enhancement for members of this community may enable health workers to more accurately target potential users when putting forth intervention efforts. Future research should aim to gain a better understanding into the role that methamphetamine plays in non-sexual contexts, particularly among gay and bisexual men who may not be part of the club "scene." The relationship between attitudes towards methamphetamine and other drugs, particularly cocaine, among gay and bisexual men should be explored. PMID- 17398041 TI - Identification of five mouse mu-opioid receptor (MOR) gene (Oprm1) splice variants containing a newly identified alternatively spliced exon. AB - The mouse mu-opioid receptor gene, Oprm1, currently contains 18 recognized alternatively spliced exons [Doyle, G.A., Sheng, X.R., Lin, S.S.J., Press, D.M., Grice, D.E., Buono, R.J., Ferraro, T.N., Berrettini, W.H., 2007. Identification of three mouse mu-opioid receptor (MOR) gene (Oprm1) splice variants containing a newly identified alternatively spliced exon. Gene 388 (1-2) 135-147, in press (doi:10.1016/j.gene.2006.10.017). Electronic publication 2006 November 1] that generate 27 splice variants encoding at least 11 morphine-binding isoforms of the receptor. Here, we identify five MOR variants that contain an as yet undescribed exon (exon 19) of the gene, and we provide evidence that these MOR splice variants are expressed in the C57BL/6 and DBA/2 mouse strains. Three splice variants, MOR-1Eii, MOR-1Eiii and MOR-1Eiv, encode the MOR-1E isoform and contain the newly identified exon 19 in their 3' untranslated regions. The fourth splice variant encodes a novel mu-opioid receptor isoform, MOR-1U, and contains exon 19 in its coding region. The cytoplasmic tail of the putative MOR-1U isoform contains a putative nuclear localization signal encoded by the sequence of exon 19. Exon 19 appears to be conserved in the rat, but not in humans. In mouse and rat Oprm1, exon 19 is located between described exons 7 and 8. We also report the cloning of the "full-length" MOR-1T splice variant [Kvam, T.-M., Baar, C., Rakvag, T.T., Kaasa, S., Krokan, H.E., Skorpen, F., 2004. Genetic analysis of the murine mu-opioid receptor: increased complexity of Oprm1 gene splicing, J. Mol. Med. 82 (4) 250-255] that encodes MOR-1 and contains the newly identified exon in its 3' UTR. RT-PCR analysis suggests that splice variants MOR-1Eii, MOR-1Eiii, MOR-1Eiv, MOR-1T and MOR-1U are expressed in all brain regions analyzed (cortex, cerebellum, hypothalamus, thalamus and striatum). These exon 19-containing splice variants add to the growing complexity of the mouse Oprm1 gene. PMID- 17398042 TI - Factors affecting chromatin stability of bovine spermatozoa. AB - The structural stability of transcriptionally inert paternal chromatin is of vital importance for the fertilization process and early embryonic development. Accordingly, a series of eight experiments were conducted during a 7-month period to investigate: (1) effects of bull breed, individuality, successive ejaculations, semen quality characteristics (SQC), semen dilution rates and hypothermic storage of semen in a Tris-egg yolk extender on incidence of sperm nuclear chromatin instability (NCI), and (2) effects of the interaction between variation of NCI within a frozen ejaculate and variation of oocytes quality due to maturation time and/or season on the efficiency of in vitro embryo production (IVEP). Semen samples were collected once a week from six bulls using an AV and only ejaculates (n=220) of >0.30x10(9) sperm/ml and >or=60% motility were used. NCI was measured by: (1) detection of lysine-rich histones in sperm chromatin using aniline blue staining, (2) sperm susceptibility to acid-induced nuclear DNA denaturation in situ using acridine orange test, and (3) sperm susceptibility to nuclear chromatin decondensation (NCD). Bovine oocytes (n=695) were matured in vitro for 18 or 24 h, fertilized after sperm selection through a swim-up procedure and cultured for 72 h. The results showed that the 2nd ejaculates were superior to the 1st ones with respect to chromatin stability. Dilution of semen to 49.67+/-8.56x10(6) sperm/ml (1:19) decreased resistance of sperm to NCD. Cooling of semen had no significant effect on chromatin stability. Cryopreservation of semen augmented sperm vulnerability to DNA denaturation. Improvement of SQC (semen volume, sperm motility, velocity, viability and morphological normalcy) was generally concomitant with increase of sperm resistance to NCI. While Blonde d'Aquitaine bulls had a resistance to NCD higher than Limousine bulls in fresh semen, the former showed a greater susceptibility to DNA denaturation than the latter in cooled semen. Individuality significantly influenced NCI. The variability of NCI within a frozen ejaculate affected efficiency of IVEP. Significant negative correlations were observed between incidence of NCI and both fertilization rate and developmental capacity of embryos after maturation of oocytes for 18 h. The significant variation in IVEP traits due to season was independent of the effect of sperm chromatin instability. PMID- 17398043 TI - Interaction of omapatrilat with FD&C Blue No. 2 lake during dissolution of modified release tablets. AB - The purpose of this study was to identify the mechanism(s) of omapatrilat degradation observed during dissolution from modified release (MR) tablet formulations containing colorants. The tablets were manufactured by a dry granulation process employing roller compaction. The colorants were added intragranularly and included red and yellow iron oxides and FD&C Blue No. 2 lake and dye. Dissolution studies in pH 6 or 6.8 media do not indicate any omapatrilat degradation in the absence of colorants. In the presence of colorants the degradation rate of omapatrilat in pH 6.8 media was in the following order: blue lake>blue lake+yellow iron oxide>yellow and red iron oxides. Higher degradation was observed with MR tablets formulated with indigo carmine (dye) as opposed to tablets formulated with aluminum oxide or aluminum hydroxide (dye substrate portion of lake). The inclusion of tartaric acid and the photostabilizer, uric acid, in omapatrilat MR tablets containing the blue lake reduced the degradation significantly. The dissolution instability observed at pH 6.8 in the MR tablet formulated with FD&C Blue No. 2 Lake was attributed to the dye component of the lake. The instability was more pronounced at higher pH and in the absence of a photostabilizer. PMID- 17398044 TI - Buserelin acetate microparticle dispersion effects drug release and plasma E(1) levels. AB - We investigated the effect of different dispersion methods on release behavior and efficacy onset following microparticle administration of buserelin acetate (BA) sustained-release injection. In this in vitro release study, the initial dispersion of BA increased with increased stirring speed (p<0.01). Stability of BA was studied over 7 days after BA release. The initial BA release rate was higher (p<0.01) after a 1-min vibration dispersion method (VDM) using a test tube mixer (2000 rpm) compared with the standard dispersion method (SDM) by hand. Without shaking, powder aggregation was observed, and BA release was lower than in either the SDM or VDM methods. In this study using 4-week-old Sprague-Dawley female rats, the initial plasma estrone (E(1)) concentrations were lower (p<0.05) in the VDM method than in the SDM method. Observations by optical microscope and scanning microscope showed no change in microparticle shape or distribution of size induced by SDM, VDM or the ultrasonication dispersion method. These results suggest that different dispersion methods do not change the shape and distribution of microparticle size, but clearly change the BA release rate and the transition in plasma E(1) concentrations that can affect drug efficacy. PMID- 17398045 TI - Itraconazole IV nanosuspension enhances efficacy through altered pharmacokinetics in the rat. AB - The goal of this research was to evaluate an intravenous itraconazole nanosuspension dosage form, relative to a solution formulation, in the rat. Itraconazole was formulated as a nanosuspension by a tandem process of microcrystallization followed by homogenization. Acute toxicity, pharmacokinetics, and distribution were studied in the rat, and compared with a solution formulation of itraconazole. Efficacy was studied in an immunocompromised rat model, challenged with a lethal dose of either itraconazole sensitive or itraconazole-resistant C. albicans. Itraconazole nanosuspension was tolerated at significantly higher doses compared with a solution formulation. Pharmacokinetics of the nanosuspension were altered relative to the solution formulation. C(max) was reduced and t(1/2) was much prolonged. This occurred due to distribution of the nanosuspension to organs of the monocyte phagocytic system (MPS), followed by sustained release from this IV depot. The higher dosing of the drug, enabled in the case of the nanosuspension, led to higher kidney drug levels and reduced colony counts. Survival was also shown to be superior relative to the solution formulation. Thus, formulation of itraconazole as a nanosuspension enhances efficacy of this antifungal agent relative to a solution formulation, because of altered pharmacokinetics, leading to increased tolerability, permitting higher dosing and resultant tissue drug levels. PMID- 17398046 TI - Immobilization and bioactivity of glucose oxidase in hydrogel microspheres formulated by an emulsification-internal gelation-adsorption-polyelectrolyte coating method. AB - The purpose of this study was to develop a novel microsphere formulation of glucose oxidase (GOX) with high drug loading, encapsulation efficiency and bioactivity. GOX was encapsulated in alginate/chitosan microspheres (ACMS) using an emulsification-internal gelation, followed by GOX adsorption and polyelectrolyte coating method. The factors influencing GOX loading, encapsulation efficiency and activity of the loaded GOX were investigated. The resultant ACMS in wet state were spherical with a mean diameter of about 138 microm. GOX loading was found to be pH dependent. High GOX loading and encapsulation efficiency were achieved when the pH of the adsorption medium was lower than the isoelectric point (pI) of GOX. GOX loading and encapsulation efficiency increased with increasing GOX concentration in the loading solution, but decreased with increasing chitosan concentration in the coating solution. The activity of loaded GOX increased and then decreased with increasing chitosan concentration. The activity of GOX in ACMS was maintained and showed sustained production of H(2)O(2) as compared to free GOX. Around 90% of the original activity of immobilized GOX remained after lyophilization and storage at -20 degrees C for a month. These results suggest that the ACMS and the fabrication method are suitable for microencapsulation of proteins like GOX. PMID- 17398047 TI - The effects of high shear blending on alpha-lactose monohydrate. AB - alpha-Lactose monohydrate is an important pharmaceutical excipient used extensively in dry powder inhaler (DPI) formulations. The ways in which a high shear blending process affect this material have been investigated and important process parameters have been identified. Total energy input (kJ/kg), blade design and the conditions in which lactose was stored prior to blending were found to have the most significant effect on the apparent particle size distribution of the processed material, which may subsequently affect the performance of DPI formulations. The power conditions used during blending, equipment temperature and humidity of the headspace above the powder were found to be less important in this respect. Additionally, it was found that high energy blending could induce changes in the water sorption characteristics of the material, although the formation of amorphous material could not be confirmed. PMID- 17398048 TI - Development of carbonyl iron/ethylcellulose core/shell nanoparticles for biomedical applications. AB - A reproducible method for the preparation of mixed colloidal nanoparticles, consisting of a magnetic carbonyl iron nucleus and a biocompatible ethylcellulose latex shell, is described in this article. The heterogeneous structure of the particles can confer them both the possibility of being used as drug delivery systems and the responsiveness to external magnetic fields, allowing a selective guidance of drug molecules to specific target tissues without a concurrent increase in its level in healthy tissues. The preparation method is based on an emulsion solvent evaporation process. A complete physicochemical characterization of the composite particles was carried out, and this preliminary investigation showed that the surface behavior of the core/shell particles is similar to that of bare ethylcellulose particles. This was confirmed, in particular, by zeta potential determinations as a function of pH and ionic strength. This fact points to the ethylcellulose shell efficiently coating carbonyl iron, and leading to composite particles which, from the electrokinetic point of view, are almost indistinguishable from latex. The thermodynamic analysis agrees with the electrokinetic one in suggesting that the coverage has been complete, since the components of the surface free energy of mixed particles coincide almost exactly with those corresponding to the cellulose-based pseudolatex. Moreover, the hydrophilic nature of carbonyl iron is modified and the particles become hydrophobic, just like the latex, when they are covered by ethylcellulose. The magnetic behaviors of the carbonyl iron and composite particles were also checked, and the similarities between both types of particles were demonstrated, except that the polymeric shell reduces the magnetization of the sample. PMID- 17398049 TI - Synthesis of poly(sebacic anhydride)-indomethacin controlled release composites via supercritical carbon dioxide assisted impregnation. AB - Poly(sebacic anhydride), PSA and indomethacin drug composite (DC) formulations were prepared using supercritical CO(2) (sc-CO(2)) aided mixing. The effect of the experimental temperature and sebacic acid purity on the physical properties of PSA-indomethacin DCs was investigated using a range of analytical techniques. The nature of the PSA-indomethacin interaction in composites after processing in sc-CO(2) under various conditions was investigated using differential scanning calorimetry (DSC), Fourier transform infrared (FTIR) spectroscopy, and powder X ray diffraction (XRD) methods, respectively. The results indicate that processing at 130 degrees C of a 4:1 (w/w) ratio PSA-indomethacin mixture, renders the indomethacin amorphous and dispersed within the polymer matrix. The primary interaction between PSA and indomethacin appears to be hydrogen bonding between the carboxylic acid OH of indomethacin and the carbonyl group of PSA. In vitro dissolution studies revealed that the processed composites exhibit a substantially enhanced dissolution rate compared to the physical mixtures. Also, through the control of experimental conditions, the initial burst effect of the drug release was largely alleviated. Instead, the erosion of PSA (zero order degradation) became the dominant factor in controlling the drug release rate. PMID- 17398050 TI - Hypoglycemic effect of Tournefortia hirsutissima L., on n-streptozotocin diabetic rats. AB - The hypoglycemic effect of aqueous and butanolic extracts from Tournefortia hirsutissima (Boraginaceae) was determined on neonatal induced streptozotocin diabetic rats (n-STZ). Oral administration of water extracts at doses of 20 and 80mg/kg, and butanolic extracts (8 and 80mg/kg) significantly lowered the plasma glucose levels in diabetic rats within 3h. Glibenclamide was used as reference and showed similar hypoglycemic effect. Our results support the traditional use of the plant as a hypoglycemic agent; we observe a dose-dependent action of the extracts. HPLC analysis confirmed that the aqueous and butanolic extracts had the same chemical composition. PMID- 17398051 TI - Anticonvulsant activity of Cotyledon orbiculata L. (Crassulaceae) leaf extract in mice. AB - The anticonvulsant activity of Cotyledon orbiculata L. (Crassulaceae) was investigated by studying the effects of both aqueous and methanol extracts of the plant species on seizures induced by pentylenetetrazole, bicuculline, picrotoxin and N-methyl-dl-aspartic in mice. Aqueous extract of Cotyledon orbiculata (50 400mg/kg, i.p.) and methanol extract (100-400mg/kg, i.p.) significantly prolonged the onset of tonic seizures induced by pentylenetetrazole (95mg/kg, i.p.). Methanol extract (400mg/kg, i.p.) also significantly reduced the incidence of the seizures. One hundred to two hundred milligrams/kilogram (i.p.) of aqueous extract of Cotyledon orbiculata significantly delayed the onset of the tonic seizures induced by bicuculline (40mg/kg, i.p.), picrotoxin (12mg/kg, i.p.) and N methyl-dl-aspartic acid (NMDLA, 400mg/kg, i.p.). Similarly, methanol extract (100 400mg/kg, i.p.) significantly delayed the onset of the tonic seizures induced by bicuculline (40mg/kg, i.p.) and picrotoxin (12mg/kg, i.p.) while 100mg/kg (i.p.) significantly delayed the onset of N-methyl-dl-aspartic acid (NMDLA, 400mg/kg, i.p.)-induced seizures. Methanol extract (200mg/kg, i.p.) significantly reduced the incidence of the seizures induced by bicuculline (40mg/kg, i.p.). Phenobarbitone (12mg/kg, i.p.) and diazepam (0.5mg/kg, i.p.) effectively antagonized only seizures induced by PTZ (95mg/kg, i.p.), bicuculline (40mg/kg, i.p.) and picrotoxin (12mg/kg, i.p.). Phenytoin (30mg/kg, i.p.) did not affect any of the seizures to any significant extent. The data obtained suggest that both aqueous and methanol extracts of Cotyledon orbiculata have anticonvulsant property and may probably be affecting both gabaergic and glutaminergic mechanisms to exert its effect. The phytochemical analysis carried out revealed the presence of cardiac glycosides, saponins, tannins, reducing sugar and triterpene steroids in the plant extract. PMID- 17398053 TI - Evaluation of lymph node metastases of breast cancer using ultrasmall superparamagnetic iron oxide-enhanced magnetic resonance imaging. AB - BACKGROUND: We assessed the utility of enhanced magnetic resonance imaging (MRI) using ultrasmall superparamagnetic iron oxide (USPIO) in the evaluation of axillary lymph node metastases in patients with breast cancer. STUDY DESIGN: MR examination of the axilla was performed before and 24-36 h after USPIO administration for patients with stage II or III breast cancer. Diagnostic performance was compared using size criteria (metastasis was defined when short axis diameter >5 or >10mm) or morphologic criteria on conventional MRI, the combined study of USPIO precontrast and postcontrast images, and USPIO postcontrast study alone. RESULTS: A total of 622 nodes (503 metastatic and 119 nonmetastatic nodes) were dissected from 33 patients. The results of conventional MRI for nodes >5mm were 59.1% sensitivity, 86.7% specificity, and 80.4% overall accuracy. Results for nodes >10mm were 15.7% sensitivity, 99.2% specificity, and 80.2% overall accuracy. Results based on morphology were 36.5% sensitivity, 94.1% specificity, and 81.0% overall accuracy. The results of the combined study of USPIO precontrast and postcontrast images were 86.4% sensitivity, 97.5% specificity, 91.1% positive predictive value, 96.1% negative predictive value, and 95.0% overall accuracy. The results of USPIO postcontrast images alone were 84.7% sensitivity, 96.8% specificity, and 94.0% overall accuracy. Patient-based results of postcontrast USPIO study alone were 100.0% sensitivity, 80.0% specificity, and 93.9% overall accuracy. CONCLUSIONS: USPIO postcontrast study alone was useful in the assessment of axillary lymph node metastases in patients with breast cancer. PMID- 17398054 TI - Editorial. Digestive system radiology in the Czech Republic. PMID- 17398055 TI - Nateglinide quantification in rabbit plasma by HPLC: optimization and application to pharmacokinetic study. AB - A rapid, simple, and sensitive HPLC method with UV detection was developed and validated for the determination of nateglinide (NTG) from rabbit plasma. The retention behavior of NTG and gliclazide (GLZ, internal standard-IS) as a function of mobile phase pH, composition and flow rate was investigated. Separation was developed on a reverse-phase C(18) column (250 mm x 4.6mm i.d., 5 microm particle size), using a mixture of acetonitrile (ACN):10mM phosphate buffer (PBS, pH 3.0) in the ratio of 70:30(%v/v) at a flow rate of 1.0 ml/min with UV detection at 203 nm within 8 min, and quantified based on drug/IS peak area ratios. The plasma samples were prepared by a simple deproteinization with a mixture of methanol and acetonitrile, yielding more than 97.86% extraction efficiencies. The calibration curve was linear (correlation coefficient of 0.9984) in the concentration range of 10-2500 ng/ml. The limit of detection (LoD) and limit of quantitation (LoQ) were found to be 2.91 and 9.70 ng/ml, respectively. Both the intra-day and inter-day precisions at four tested concentrations were below 1.32% R.S.D. The present method was selective enough to analyze NTG in rabbit plasma without any tedious sample clean-up procedure and was successfully applied for estimating the pharmacokinetic parameters of NTG following oral administration of a single 15 mg NTG to white albino rabbits. PMID- 17398056 TI - Total percutaneous access for endovascular aortic aneurysm repair ("Preclose" technique). AB - OBJECTIVE: Percutaneous access during endovascular aneurysm repair has been difficult owing to the large size of the delivery catheters. This study reports a single-center experience of totally percutaneous access during endovascular abdominal and thoracic aortic repairs using the Preclose Proglide device (Abbott Vascular, Redwood City, Calif). METHODS: Between December 2004 and August 2006, 262 endovascular aortic aneurysm repairs were performed. Percutaneous access was used for the introduction of 12F to 24F sheaths (4.4-mm to 8.6-mm outer diameter). The technique involved deployment of two Proglide devices before insertion of the sheath ("Preclose" technique) with the sutures left extracorporeally for closure after conclusion of the procedure. A prospectively maintained endovascular database and medical records were retrospectively reviewed. Rates of technical success, failure modes, and the overall duration of the endovascular repair compared with a similar cohort using open femoral exposures were examined. RESULTS: A total of 559 Proglide devices were used to close 279 femoral arteries, and 175 (63%) required the insertion of 18F to 24F sheaths. There were 16 failures, mainly due to obesity, device malfunction, severe calcific disease, and faulty arterial punctures, for a technical success rate of 94.3%. The success rates for 12F to 16F size sheaths were significantly higher than for the larger 18F to 24F sheaths (99.0% vs 91.4%, P<.01). For both endovascular abdominal (EVAR) and thoracic (TEVAR) aortic repairs, the Preclose technique resulted in shorter overall procedure times compared with a similar cohort in which open femoral exposures were used (EVAR, 115 vs 128 min, P<.001; TEVAR, 80 vs 112, P=.019). Despite this reduction of procedure time, the savings on the cost of operating room time was negated by the cost of the Proglide devices ($295 per device). CONCLUSIONS: Percutaneous access for endovascular aortic repair is safe and feasible using the Proglide device. Although the success rates are higher for smaller size sheaths, successful closures may be obtained for up to 24F sheaths. Percutaneous access may result in shorter overall procedure times and potentially lower operating room costs, but this appears to be offset by the cost of the closure devices. PMID- 17398057 TI - Five-year report of a multicenter controlled clinical trial of open versus endovascular treatment of abdominal aortic aneurysms. AB - OBJECTIVE: Compare long-term results of endovascular treatment and standard open repair of abdominal aortic aneurysms in a multicenter, concurrent-controlled trial. METHODS: 334 subjects were treated with standard open repair (control, n = 99) or the original EXCLUDER Bifurcated Endoprosthesis (test, n = 235). Five-year clinical evaluations and corelab radiographic results are analyzed. RESULTS: Overall and aneurysm-related survival are similar. There have been ten open conversions, most frequently for enlarging sacs without endoleak. Two patients died after conversion. Including reinterventions and complications of reinterventions as adverse events, there is significant, persistent long-term reduction in major adverse events. At 5 years, corelab reported 0% limb narrowing, 0% trunk migration, 0% component (contralateral leg, aortic extender, and iliac extender) migration, 0% fracture, endoleak in 3% (2 type II/68), and aneurysm growth (>5 mm compared to baseline) in 38% (30/78) of the test group. There are no aneurysm ruptures in either test or control group. CONCLUSIONS: After 5 years follow-up, endovascular repair is a safer and effective treatment compared with open surgical repair for abdominal aortic aneurysms. Major adverse events are less frequent with the endograft despite the need for late reinterventions. Aneurysm expansion is observed in nearly two-fifths of patients but is not associated with endoleak or aneurysm rupture. Multicenter clinical trials are evaluating a newer version of this device designed to avoid this high rate of sac expansion. PMID- 17398058 TI - Cigarette smoking increases aortic dilatation without affecting matrix metalloproteinase-9 and -12 expression in a modified mouse model of aneurysm formation. AB - OBJECTIVE: The development of abdominal aortic aneurysms (AAA) is presumed to result from multiple genetic and environmental factors, with exposure to tobacco smoke the single largest known factor predisposing to aneurysm growth. We have attempted to adapt the elastase-perfused animal model to determine whether tobacco exposure can lower the threshold of aortic injury necessary for AAA development. METHODS: Adult C57BL/6 mice underwent transient perfusion of the infrarenal aorta with an active solution of elastase: high-dose (HDE, 0.19 U/mL, n=9), standard-dose (SDE, 0.16 U/mL, n=21) or low-dose (LDE, 0.07 U/mL, n=24). Control animals (n=24) were treated with heat inactivated elastase (HIE). Twenty LDE perfused mice were exposed to cigarette smoke (LDE-S) beginning 2 weeks before perfusion and continuing until aortic harvest. Aortic diameter (AD) was measured preperfusion, postperfusion, and at harvest on day 14. AAA was defined as %DeltaAD>or=100% between preperfusion and harvest. Aortas from each group (except HDE) were analyzed for matrix metalloproteinase-9 (MMP-9) and MMP-12 expression by real-time polymerase chain reaction normalized to glyceraldehyde-3 phosphate dehydrogenase. RESULTS: All SDE mice developed large AAA by %DeltaAD (189.3%+/-16.9%, mean+/-standard error of the mean), but control mice had only a small dilatation (69.7%+/-3.7%, P<.01). Higher doses of elastase did not produce larger aneurysms in HDE mice. In contrast, only 63% of LDE mice showed aneurysmal dilatation, and these were significantly smaller (104.3%+/-4.2%, P<.01). When exposed to cigarette smoke, LDE animals developed significantly larger aneurysms (%DeltaAD, 134.5%+/-7.9%, P=.0021). There was no difference in normalized aortic MMP-9 and MMP-12 expression between elastase doses or between smoke-exposed and unexposed animals. Histologic analysis revealed that smoking increased the extent of aortic elastin degradation when compared with LDE-S animals. CONCLUSION: Aneurysm development in the elastase model is dependent on the quantity of active elastase infused. Exposure of animals to tobacco smoke after a relatively minor aortic elastase injury produces increases in elastin degradation and aneurysm size without affecting MMP-9 or MMP-12 expression. To our knowledge, this is the first demonstration in an animal model that smoking can act as a synergistic factor in AAA development. Further understanding of the relationship between smoking and AAA in this model may help unveil the pathophysiologic pathways involved between cigarette smoke and AAAs. PMID- 17398059 TI - Pectoralis minor obstruction of the axillary vein: report of six patients. AB - PURPOSE: Although the usual site of nonthrombotic venous obstruction of the upper extremity is the subclavian vein, other sites may be the cause of such obstruction. This study describes the diagnosis and treatment of six patients with partial axillary vein obstruction by the pectoralis minor muscle, a condition that can mimic subclavian vein obstruction. METHODS: A chart review of patients undergoing pectoralis minor tenotomies (PMT) between 2004 and 2006 revealed six patients (3 men and 3 women), aged 17 to 39, who underwent seven PMT procedures for symptoms of arm swelling, cyanosis, and pain or tightness. Diagnosis was suggested by history and physical examination and was confirmed by dynamic venography. Patients with paresthesia suggesting associated neurogenic pectoralis minor compression were given a pectoralis minor muscle block. As an outpatient, PMT was initially performed with an infraclavicular approach but later through the transaxillary route. Follow-up was by phone interview in five patients and a physical examination in one. RESULTS: Venography demonstrated axillary vein compression under the pectoralis minor, which was more significant than the minor degree of subclavian vein compression seen on the same venogram. Follow-up was 1.5 years to 10 years in three patients and 3 months in the other three. All six patients experienced good-to-excellent relief of all symptoms. There were no surgical complications. CONCLUSION: Axillary venous obstruction by the pectoralis minor must be distinguished from subclavian vein obstruction, which presents with similar symptoms. PMT is a simple, risk-free, outpatient procedure that has produced uniformly good results. PMID- 17398060 TI - Tumour characteristics predictive of survival following resection for ductal adenocarcinoma of the head of pancreas. AB - AIMS: We have maintained a highly conservative policy in selecting patients with carcinoma of the head of pancreas for resection. This has been based on tumour size, evidence of lymph node involvement or local invasion outside of the gland at laparotomy, laparoscopy or CT imaging. This study investigated our survival rates following pancreatic resection and examined clinicopathological predictors of survival. METHODS: Sixty-two consecutive patients undergoing pancreatic resections for malignancy were identified from 1999 onwards. Thirty-three underwent resection for pancreatic ductal adenocarcinoma and were included in our analysis, the remainder included resections for ampullary adenocarcinoma (n=20) or other malignancies (n=9). Patient, tumour and operative characteristics were analysed to assess predictors of survival following resection (Kaplan-Meier survival curves). RESULTS: Median survival following resection for ductal pancreatic adenocarcinoma was 54 months (ampullary adenocarcinomas achieved a median survival of 62 months) and thirty-day mortality was 2.7% (n=1). Survival was not associated with any demographic or intraoperative factors, such as blood loss, operative duration or anaesthetic technique. Survival curves were significantly worse when perineural or vascular invasion was evident histologically (p=0.023 and 0.0023 respectively). Patients with positive lymph nodes had a significantly shorter survival (p=0.0030) especially when lymph node status was expressed as a percentage of total lymph node yield. If more than 20% of retrieved lymph nodes were positive for tumour, this was a clear predictor of survival (p<0.0001). A positive resection margin was also associated with shortened survival (p=0.0291). CONCLUSION: Despite the advances made in the management of pancreatic cancer, tumour biology still dictates long-term survival. A highly selective surgical approach to the management of these patients results in good long-term survival. PMID- 17398061 TI - [Maternal and neonatal effects of remifentanil for general anaesthesia for Caesarean delivery]. AB - OBJECTIVE: Opioids are routinely omitted at the induction of general anesthesia for Caesarean delivery because of the risks of respiratory neonatal depression. The short-acting opioid remifentanil may afford advantages at the induction and surgical stimulation, without subsequent neonatal depression. PATIENTS AND METHODS: In this double-blinded study, 40 at term women undergoing elective Caesarean section and requiring general anaesthesia were allocated randomly to receive either remifentanil (0,5 microg/kg) at the induction of anaesthesia (G1, n=20) or placebo (G2, n=20). Induction of anaesthesia was performed with propofol 2 mg/kg and succinylcholine 1 mg/kg. Anaesthesia was maintained with nitrous oxide in oxygen (50/50%, v/v), propofol (100 microg/kg/min), remifentanil (0.2 microg/kg/min) and atracurium. Neonates were assessed by using Apgar scores, possible respiratory depression, with or without ventilation in the mask or intubation and umbilical cord blood gas (artery: UA and vein: UV). Values are expressed as mean values +/-SD. Pearson's Chi squared and t-test were used for statistical analysis P<0.05 was considered significant. RESULTS: Maternal systolic pressure, mean pressure and heart rate were significantly higher in G1 at induction. Apgar scores, heart and respiratory rate were similar between groups. Seven episodes of respiratory depressions were noted (3 in G1, 4 in G2). Five neonates required only brief assisted ventilation by face-mask (2 in G1, 3 in G2). CONCLUSION: Remifentanil (0.5 microg/kg) at the induction of anaesthesia in elective Caesarean section under general anaesthesia can be used without subsequent neonatal depression. However, we believe that further research is necessary to extrapolate these results to a pregnancy carrying an acutely distressed foetus. PMID- 17398063 TI - [Death during adenoidectomy]. PMID- 17398064 TI - [Human dirofilariasis: 3 cases in the south of France]. AB - INTRODUCTION: Dirofilariasis is a zoonosis usually found in dogs and cats. It is rare in humans, who are dead-end hosts for the parasite. CASES: We report 3 cases of subcutaneous dirofilariasis due to Dirofilaria repens, contracted in the south of France (Alpes-Maritimes and Corsica). In the first two cases, the dirofilariasis manifested as lymph node enlargement; in the third case, lung disease suggested a systemic diffusion of microfilariae. DISCUSSION: Dirofilaria repens dirofilariasis is due to the transmission of microfilariae by some mosquito bites (Aedes, Culex, Anopheles, Mansonia, Psorophora and Taeniorhynchus). Usually only one larva develops, producing an immature adult worm inside a node. Ultrasound examination may suggest the parasitic origin of the lesion. It is treated surgically, by excision, without chemotherapy. Very rarely, an adult worm may mature and produce systemic diffusion of microfilariae. The nodule in the third case contained a gravid adult female worm but we found no microfilariae. Dirofilariosis can present problems in diagnosis and treatment. It must be considered in patients with an isolated nodule. PMID- 17398065 TI - [Stop citing fraudulent and duplicate articles]. PMID- 17398066 TI - The haemolytic effect of verapamil on erythrocytes exposed to varying osmolarity. AB - The haemolytic effect of verapamil on red blood cells (RBCs) exposed to varying osmolarity was investigated. The experimental approach used a modified red cell haemolysis assay with concentrations of verapamil ranging from 50-1500 microM compared to drug free controls. The time-course of haemolytic effects was also investigated. We also briefly determined the haemolytic effects of verapamil in Ca2+-free conditions (with added EGTA). In conditions representing decreasing osmolarity (dilution from 140-0 mM NaCl) there was a significant increase in erythrocyte haemolysis that was also dependent on verapamil concentration (ANOVA, p<0.05). The red cells also showed a significantly increased rate of haemolysis over 5 h with increasing verapamil concentration (ANOVA, p<0.05). The degree of RBC hypotonic haemolysis was significantly increased in a Ca2+-free medium (+EGTA) compared to normal saline and this effect was exacerbated by additions of verapamil (ANOVA, p<0.05). Overall the data suggested that verapamil can cause haemolysis of RBCs in a predictable time- and concentration-dependent manner, and that verapamil increases the fragility of the erythrocytes further during hypotonic osmotic stress and Ca2+-free conditions. The mechanism of verapamil dependent haemolysis could be directly related to the observed biphasic concentration-effect and could consequently involve several ion transport pathways. PMID- 17398068 TI - Knowledge and use of folic acid for prevention of birth defects amongst Honduran women. AB - PURPOSE: To characterize determinants of folic acid (FA) use among women of reproductive age and patient education practices of health care professionals in one region of Honduras. METHODS: 508 female outpatients and 128 health workers were interviewed in six primary care clinics in Honduras. Results were analyzed using univariate and multivariate regression models. RESULTS: The survey showed that 45% patients were familiar with FA. Of that number, 30% knew appropriate timing of consumption and 25% reported proper pre-natal supplementation. Increasing education was strongly correlated with knowledge of folic acid function (OR=252.52, P<0.0001) and actual use (OR=12.65, P<0.000). Age is associated with knowledge of proper timing of FA usage (OR=3.94, P<0.01). Most women learned about FA from medical professionals, but only half of providers educate their female patients about FA. CONCLUSIONS: Healthcare providers should remember to discuss the role of folic acid with their patients, particularly those of low education and at the extremes of reproductive age. While long-term efforts to develop fortification programs continue, interim Honduran health campaigns to increase proper consumption of folic acid should target these particularly vulnerable populations. PMID- 17398067 TI - Maternal flaxseed diet during pregnancy or lactation increases female rat offspring's susceptibility to carcinogen-induced mammary tumorigenesis. AB - Flaxseed contains several dietary components that have been linked to low breast cancer risk; i.e., n-3 polyunsaturated fatty acids (PUFAs), lignans and fiber, but it also contains detectable levels of cadmium, a heavy metal that activates the estrogen receptor (ER). Since estrogenic exposures early in life modify susceptibility to develop breast cancer, we wondered whether maternal dietary intake of 5% or 10% flaxseed during pregnancy or lactation (between postpartum days 5 and 25) might affect 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumorigenesis in the rat offspring. Our data indicated that both in utero and postnatal 5% and 10% flaxseed exposures shortened mammary tumor latency, and 10% flaxseed exposure increased tumor multiplicity, compared to the controls. Further, when assessed in 8-week-old rats, in utero 10% flaxseed exposure increased lobular ER-alpha protein levels, and both in utero and postnatal flaxseed exposures dose-dependently reduced ER-beta protein levels in the terminal end buds (TEBs) lobules and ducts. Exposures to flaxseed did not alter the number of TEBs or affect cell proliferation within the epithelial structures. In a separate group of immature rats that were fed 5% defatted flaxseed diet (flaxseed source different than in the diets fed to pregnant or lactating rats) for 7 days, cadmium exposure through the diet was six-fold higher than allowed for humans by World Health Organization, and cadmium significantly accumulated in the liver and kidneys of the rats. It remains to be determined whether the increased mammary cancer in rats exposed to flaxseed through a maternal diet in utero or lactation was caused by cadmium present in flaxseed, and whether the reduced mammary ER-beta content was causally linked to increased mammary cancer risk among the offspring. PMID- 17398069 TI - Identification of the breast edge using areas enclosed by iso-intensity contours. AB - The segmentation of a mammogram into background and breast is a crucial first step in the computer aided diagnosis of mammograms that has the advantage of simplifying further processing of the image (by eliminating the background) and also provides a reference for the alignment of views when two views are being compared. A novel method of segmenting the breast from the background by analysing the area enclosed by iso-intensity contours is presented. Results are evaluated by comparison with manual borders drawn by three radiologists for a set of 25 mammograms. The effect of different pre-processing methods, on the accuracy of automated borders, is investigated. The best methods yielded average root-mean square differences between the manual and automated iso-intensity borders of 3.0+/-0.3 mm for the image set containing clear breast edges and 4.8+/-0.5 mm for the image set containing indistinct breast edges compared to 5.1+/-0.8 and 7.9+/ 0.9 mm for the two data sets with no pre-processing. Results are also compared to those obtained from global thresholding. PMID- 17398070 TI - Cot/Tpl2 and PKCzeta cooperate in the regulation of the transcriptional activity of NFATc2 through the phosphorylation of its amino-terminal domain. AB - Nuclear factor of activated T cells (NFAT) plays a prominent role in gene transcription during the immune response. Growing evidence demonstrates the implication of inducible phosphorylation of the transactivation domain (TAD) of NFAT in transcriptional activation of genes. We have analyzed the regulation of NFATc2 activation by Cot/Tpl2 and protein kinase C zeta (PKCzeta) in T cells. Our results show that PKCzeta and Cot/Tpl2 cooperate in regulating the transactivation activity mediated by the amino-terminal domain of NFATc2. Neither Cot/Tpl2 kinase nor PKCzeta-mediated induction of the transactivation activity of NFATc2 was affected by cyclosporin-A treatment, supporting a calcineurin independent route in the signaling pathways mediating NFATc2 activation. Co precipitation experiments showed physical interaction among Cot/Tpl2, PKCzeta and NFATc2. Analysis of the transactivation activity of deletions in the N-terminal region of NFATc2, suggested the involvement of amino acids 52-64 of NFATc2 in the induction of its transactivating function by PKCzeta. This kinase in vitro phosphorylates NFATc2 and deletion and mutational studies identified Ser53 and Ser56 (of the SPPS motif) as substrates for PKCzeta. Thus, our results suggest that PKCzeta phosphorylation of Ser53 and Ser56 in the N-terminal TAD from NFATc2 potentiates its transactivating function in human T cells. PMID- 17398071 TI - Treatment of late lithium-pilocarpine-induced status epilepticus with diazepam. AB - PURPOSE: It is well known that status epilepticus (SE) becomes increasingly difficult to control over time. Previous studies have indicated that the electroencephalographic pattern at the time of intervention is predictive of the probability of successful treatment. However, these findings are based on studies limited to the first 2h of SE onset. Little data is available on the efficacy of treating SE at later time points. METHODS: The aim of the present study was to investigate the efficacy of diazepam (DZP) treatment given at two different phases of SE in a lithium-pilocarpine rat model: during continuous ictal discharges (CIDs, phase 3), and during late periodic epileptiform discharges (late PEDs, phase 5). Changes in cortical and hippocampal electroencephalographs (EEGs) were observed continuously during the phases of SE, as well as at 24, 36, 48, and 72h after SE onset. The effects of DZP treatment during CIDs or during late PEDs were compared to control DZP-untreated rats. RESULTS: In all three groups, hippocampal and cortical EEGs displayed five distinct phases of SE. There were no statistical differences in the duration of phases 1 and 2 among the three groups. Although DZP administration during CIDs did not terminate CIDs in most rats, it did significantly shorten the duration of phases 3 and 4 of SE. Importantly, DZP given during phase 5 successfully ended behavioral and electrographic seizures in most rats. CONCLUSIONS: Hippocampal and cortical EEGs displayed five distinct phases of SE that were similarly responsive to DZP treatment. Termination of electrographic seizures with DZP treatment was more effective in the last phase of SE (late PEDs) than in phase 3 (CIDs). These findings suggest that previous reports of DZPs decrease in efficacy over time may not be applicable to DZP treatment at 4h-post onset. PMID- 17398072 TI - Relevant dimensions of delusions: continuing the continuum versus category debate. AB - Delusions and hallucinations are common among healthy individuals but may differ from the symptoms experienced by persons with schizophrenia. It is hypothesized that specific dimensions of delusions, such as the distress associated with them, preoccupation, conviction or their content might be more relevant in distinguishing persons with from persons without schizophrenia than the mere presence of delusional beliefs. Second, it is investigated whether delusional beliefs are as closely linked to hallucinations in a non-clinical population as in persons with schizophrenia. The Peters et al. Delusions Inventory and the Launay Slade Hallucination Scale - Revised were used to assess delusional ideation and hallucinatory experiences in a population sample that reflects the general population in age, education and gender (n=359) and in persons diagnosed with life-time schizophrenia in varying stages of remission (n=53). There was a strong association of delusional ideation and hallucinatory experiences in both groups. Stepwise discriminant function revealed the distress associated with delusions as well as beliefs involving persecution and loss of control to be the most relevant aspects in distinguishing persons with from persons without schizophrenia. It is concluded that delusions should be assessed multi dimensionally, laying particular emphasis on distress and content of beliefs. PMID- 17398073 TI - Genetic diversity of Mycobacterium tuberculosis isolates at the Military Medical Academy in Ankara, Turkey. AB - Genotyping of Mycobacterium tuberculosis isolates from infected individuals can play an important role in tracking the source of infection and unraveling the epidemiology of a tuberculosis pandemic. A total of 114 M. tuberculosis isolates were genotyped by spoligotyping and results were compared with an international spoligotype database (SpoIDB4). Twenty-one spoligotyping-defined clusters including 97 patients were established, and an additional 17 unique patterns were found. Ninety-eight (85.9%) isolates belonged to previously defined shared types (STs). The ST53 (ill-defined T1 superfamily, n=31), ST41 (LAM7-TUR family, n=9), ST118 (T undefined, n=8) and ST50 (Haarlem 3, n=6) were four major clusters of our isolates. After comparison with the international SpoIDB4 database, two new intrafile clusters, ST2136 and ST2139, were created and two new interfile clusters, ST2135 and ST2140, were defined. Eight (7%) of the 17 isolates with unique patterns were found to be orphans, whereas the STs of 9 isolates had previously been deposited in the international SpoIDB4 database. In addition, two isolates with an ST pattern characteristic of the Beijing family of M. tuberculosis were found. This study shows that, although ubiquitous spoligotypes are common, several spoligotypes specific to Turkey also exist. Thus, our study may help us to better understand the spread of M. tuberculosis genotypes to Turkey. PMID- 17398074 TI - The Neff strain of Acanthamoeba castellanii, a tool for testing the virulence of Mycobacterium kansasii. AB - Virulent Mycobacterium kansasii (mainly subtype 1) may cause lung infections, whereas certain other strains (essentially subtype 3) are commonly non-pathogenic mycobacteria colonizing the human lower respiratory tract of patients. Determining the clinical significance of a strain isolated from a respiratory sample represents a major challenge for clinicians. Since some mycobacteria may use free-living amoebae as a training ground to select virulence traits, we wondered whether the Acanthamoeba castellanii amoeba could be used to determine the virulence of these intracellular bacteria. We investigated whether the growth and cytopathic effect of M. kansasii in A. castellanii correlate with the virulence of M. kansasii determined clinically and by subtyping. Pathogenic subtype 1 M. kansasii strains grew better in A. castellanii than non-pathogenic subtype 3 strains when considering both the number of bacteria per amoeba and the percentage of infected amoebae. Moreover, a subtype 3 M. kansasii strain isolated from blood culture, and thus considered pathogenic, was revealed to grow in A. castellanii similarly to pathogenic subtype 1 strains. These results suggest that amoebae may represent useful tools for testing the virulence of intracellular mycobacteria and other amoeba-resisting bacteria. This is important, since identification of novel bacterial virulence factors relies largely on in vitro assessment of virulence. PMID- 17398075 TI - New guidelines for infective endocarditis: a call for collaborative research. PMID- 17398076 TI - Continuous-infusion penicillin home-based therapy for serious infections due to penicillin-susceptible pathogens. AB - To evaluate the feasibility of continuous-infusion (CI) penicillin in the treatment of serious bacterial infections, consecutive adult patients with deep seated infections due to penicillin-susceptible pathogens were treated with CI aqueous penicillin G in a home-based programme, and their treatment outcomes were reviewed. Thirty-one patients with microbiologically proven infections completed the planned course of treatment. Twenty of 31 (65%) were followed for at least 2 months thereafter, and all remained free of relapse. One patient had fever attributable to penicillin hypersensitivity, two patients developed catheter-site infections and one patient developed catheter-related bacteraemia. Thus, CI penicillin is feasible for the home-based treatment of a variety of deep-seated infections with minimal toxicity. PMID- 17398077 TI - Serotonergic mechanisms in the basolateral amygdala differentially regulate the conditioned and unconditioned fear organized in the periaqueductal gray. AB - The amygdala is an important filter for unconditioned and conditioned aversive information. The amygdala synthesizes the stimuli input from the environment and then signals the degree of threat that they represent to the dorsal periaqueductal gray (dPAG), which would be in charge of selecting, organizing and executing the appropriate defense reaction. In this study, we examined the influence of fluoxetine microinjections (1.75 and 3.5 nmol/0.2 microL) into the lateral (LaA) and basolateral (BLA) amygdaloid nuclei on the freezing and escape responses induced by electrical stimulation of the dPAG. Freezing behavior was also measured after the interruption of the electrical stimulation of the dPAG. On the following day, these rats were also submitted to a contextual fear paradigm to examine whether these microinjections would affect the conditioned freezing to contextual cues previously associated with foot shocks. Fluoxetine injections into both amygdaloid nuclei did not change the freezing and escape thresholds, but disrupted the dPAG-post-stimulation freezing. Moreover, the conditioned freezing was enhanced by fluoxetine. Whereas 5-HT mechanisms in the amygdala facilitate the acquisition of conditioned fear they inhibit the dPAG post-stimulation freezing. However, the unconditioned fear triggered by activation of the dPAG is produced downstream of the amygdala. These findings have important implications for the understanding of the neurochemical substrates that underlie panic and generalized anxiety disorders. PMID- 17398078 TI - A preliminary fMRI study of the effects on cortical activation of the treatment of refractory auditory hallucinations with rTMS. AB - Three patients underwent fMRI during a word generation task before and after successful treatment of auditory hallucinations with rTMS and were compared to four control subjects. There was a significant increase and more normal task related brain activation in brain areas involved in language processing including left temporoparietal cortex after rTMS treatment. PMID- 17398079 TI - CSF sub-compartments in relation to plasma osmolality in healthy controls and in patients with first episode schizophrenia. AB - Preliminary evidence suggests that plasma Na(+) level/osmolality may have effects on brain morphology; thus we investigated the link between plasma osmolality and ventricle size in healthy controls and patients with first episode schizophrenia. A total of 16 patients and 28 healthy controls were examined with magnetic resonance imaging (MRI) and gave blood samples. High-resolution 3D SPGR images were obtained on a 1.5 Tesla scanner. Scalp-edited MRI volumes were used for estimates of intracranial gray, white matter and CSF. Regional changes in CSF concentration and ventricular morphology were measured. The groups did not differ in plasma osmolality, but patients had higher plasma Na(+). There were no differences in ventricle size. Controlling for plasma osmolality did not change the results. A mixed model procedure indicated a significant group effect and a significant osmolality by group interaction in ventricle measures. Healthy control group showed a significant relationship between osmolality and ventricle measures; this relationship was absent in the patients. Significant correlations between osmolality and lateral ventricle surface deformations were observed along the superior horn of the lateral ventricles in the healthy controls. These results suggest that plasma osmolality is related to ventricle size in healthy volunteers and that this physiological link is impaired in patients with first episode schizophrenia. PMID- 17398080 TI - Fronto-limbic and autonomic disjunctions to negative emotion distinguish schizophrenia subtypes. AB - Schizophrenia patients show a disconnection in amygdala-medial prefrontal cortex and autonomic arousal systems for processing fear. Concurrent functional magnetic resonance imaging [fMRI] and skin conductance recording were used to determine whether these disturbances are specific to fear, or present in response to other signals of danger. We also examined whether these disturbances distinguish a specific symptom profile. During scanning, 27 schizophrenia (13 paranoid, 14 nonparanoid) and 22 matched healthy control subjects viewed standardized facial expressions of fear, anger and disgust (versus neutral). Skin conductance responses [SCRs]were acquired simultaneously to assess phasic increases in arousal. 'With-arousal' versus 'without-arousal' responses were analysed using non-parametric methods. For controls, 'with-arousal' responses were associated with emotion-specific activity for fear (amygdala), disgust (insula) and anger (anterior cingulate), together with common medial prefrontal cortex [MPFC] engagement, as predicted. Schizophrenia patients displayed abnormally increased phasic arousal, with concomitant reductions in emotion-specific regions and MPFC. These findings may reflect a general disconnection between central and autonomic systems for processing signals of danger. This disjunction was most apparent in patients with a profile of paranoia, coupled with poor social function and insight. Heightened autonomic sensitivity to signals of fear, threat or contamination, without effective neural mechanisms for appraisal, may underlie paranoid delusions which concern threat and contamination, and associated social and interpersonal difficulties. PMID- 17398081 TI - Reply to "Cholesterol lowering and mortality: a sea of contradictions". PMID- 17398082 TI - Preparation of a novel freeze thawed poly(vinyl alcohol) composite hydrogel for drug delivery applications. AB - We describe a drug delivery system based on a physically cross-linked poly(vinyl alcohol) (PVA) hydrogel for the release of Theophylline (TH). A composite was created by freezing an aqueous solution of PVA/NaOH onto a PVA/poly(acrylic acid) substrate. This formed a strong interface and demonstrated greater physical strength than the hydrogel alone. Such systems have potential for a variety of localised controlled drug delivery applications, for example, as coatings for implantable devices. Importantly, the results suggest that a versatile synthetic platform is possible that may provide different functional materials or combination of such. The resultant samples were characterised using optical microscopy, modulated differential scanning calorimetry (MDSC) and dissolution testing. The microstructure of the gels was examined using micro-thermal analysis (microTA) which is a combination of atomic force microscopy and thermal analysis. TH was found to have an effect on the crystalline structure and dissolution showed a Fickian release, suggesting that swelling and crystallinity were the controlling mechanisms. PMID- 17398083 TI - Synergistically enhanced transdermal permeation and topical analgesia of tetracaine gel containing menthol and ethanol in experimental and clinical studies. AB - The aim of this study is to observe the synergistically enhanced percutaneous penetration and skin analgesia of tetracaine gel containing menthol and ethanol through experimental and clinical studies. Four anesthetic gels containing 4% tetracaine in carbomer vehicle named T-gel (containing no menthol or ethanol), 5%M/T-gel (containing 5% menthol), 70%E/T-gel (containing 70% ethanol, an optimal concentration for antiseptic), and 5%M+70%E/T-gel (containing both 5% menthol and 70% ethanol), respectively, were fabricated. The in vitro mouse skin permeation was investigated using a Franz diffusion cell. The mouse skin morphology was examined by a scanning electron microscope. The in vivo skin analgesic effect in mice was evaluated using the von Frey tests. To determine the efficacy of tetracaine gels for managing the pain in human volunteers, a paralleled, double blinded, placebo-controlled, randomized controlled trial design combined with verbal pain scores (VPS) was performed. The combination of menthol and ethanol (5%M+70%E/T-gel) conferred significantly higher tetracaine diffusion across full thickness mouse skin than 5%M/T-gel, 70%E/T-gel, and T-gel. The ultra structure changes of mouse skin stratum corneum treated with 5%M+70%E/T-gel were more marked compared with those of any other tetracaine gel. von Frey tests in mice showed a synergistically enhanced effect of menthol and ethanol on the analgesia of tetracaine gel. The mean VPS were significantly lower for volunteers treated with 5%M+70%E/T-gel than those receiving other gels or the EMLA cream. 5%M+70%E/T gel possessed the shortest anesthesia onset time, the longest anesthesia duration and the strongest anesthesia efficacy. Seventy percent ethanol in 5%M+70%E/T-gel not only improved the analgesic efficacy of the tetracaine gel through synergistically enhanced percutaneous permeation with menthol but also served as an antiseptic agent keeping drug application site from infection. 5%M+70%E/T-gel is a potential topical anesthesia preparation for clinical use. PMID- 17398084 TI - Dissolution test for citalopram in tablets and comparison of in vitro dissolution profiles. AB - A dissolution test for tablets containing 20 mg of citalopram was developed and validated using a reverse-phase liquid chromatographic method and this dissolution test was applied to compare dissolution profiles. The sink conditions, filters, stability of the drug and specificity on different dissolution media were tested to choose a discriminatory dissolution method which uses USP apparatus 1 with baskets rotating at 50 rpm, 900 ml of deaerated 0.1 M hydrochloric acid (HCl) as the dissolution medium. The quantitation method was also adapted and validated. The parameters of difference factor, similar factor, according to current FDA guidelines, and dissolution efficiency were employed to compare dissolution profiles. The dissolution test developed and validated was adequate for its purposes and could be applied for quality control of citalopram tablets, since there is no monograph to citalopram in tablets, this work can be used to help pharmocopoeias. PMID- 17398085 TI - Self-assembled monolayer-based piezoelectric flow immunosensor for the determination of canine immunoglobulin. AB - A simple, highly sensitive immunosensor for the direct determination of immunoglobulin (Ig) in canine serum based on a piezoelectric crystal accommodated in a flow-cell was developed and optimized. The new biosensor is also useful for discriminating between Ig subclasses present in canine serum by using specific monoclonal antibodies binding to the coated crystal. Various canine monoclonal anti-IgG were deposited onto the surface of the gold-coated crystal resonator using the self-assembly technique to form a receptor layer. The highly ordered self-assembled monolayers thus obtained provide a well-controlled surface structure and many advantages as regards sensing performance. The results obtained with the proposed immunosensor were compared with those provided by a protein A-based orientation-controlled immobilization method for the same monoclonal antibodies and also with direct physical adsorption of the antibodies. The crystal was accommodated in a flow-cell that was inserted into a buffer flowing stream in order to make resonant frequency measurements. PMID- 17398087 TI - How electron cryotomography is opening a new window onto prokaryotic ultrastructure. AB - Electron cryotomography is an emerging technology that enables thin samples, including small intact prokaryotic cells, to be imaged in three dimensions in a near-native 'frozen-hydrated' state to a resolution sufficient to recognize very large macromolecular complexes in situ. Following years of visionary technology development by a few key pioneers, several laboratories are now using the technique to produce biological results of major significance in the field of prokaryotic ultrastructure. Recent discoveries have included the surprising generality and complexity of the cytoskeleton, the connectivity of key membrane compartments, the location and architecture of large macromolecular machines such as the ribosome and flagellar motors, and the structure of some extraordinary external appendages. Through further technology development, identification of the most revealing model systems and sustained effort, electron cryotomography is poised to help resolve many fundamentally important questions about prokaryotic ultrastructure. PMID- 17398088 TI - A phase II study of epirubicin, cisplatin and capecitabine as neoadjuvant chemotherapy in locally advanced or inflammatory breast cancer. AB - AIM: To assess the efficacy and safety of epirubicin, capecitabine and cisplatin (EXC) combination therapy in locally advanced breast cancer (LABC) and investigate the predictive value of selected biomarkers. METHODS: Newly diagnosed LABC patients received four 3-weekly cycles of neoadjuvant EXC (epirubicin 60 mg/m(2) day 1; capecitabine 1000 mg/m(2) bid, days 1-14; cisplatin 60 mg/m(2)day 1) and two cycles of post-operative EXC. RESULTS: Eight (17%) of 48 patients had inflammatory breast cancer. Overall response rate was 74% (95% CI: 59-86%), including complete responses in 13% (95% CI: 5-26%). Nine (22%; 95% CI: 11-38%) of 41 patients undergoing surgery achieved pathologic complete response (pCR), giving a pCR rate of 19% (95% CI: 9-33%) in the intent-to-treat population. Haematological toxicity was manageable. The most problematic toxicities were chemotherapy-induced nausea/vomiting and hypercoagulative disorders. None of the biomarkers investigated, including HER2, predicted response. CONCLUSION: EXC showed high efficacy in LABC, with high clinical response and pCR rate. Nausea and vomiting were unexpectedly frequent, and more aggressive prophylaxis and management of these side effects is recommended in future studies of this combination. PMID- 17398089 TI - Cynara cardunculus L. alkaline pulps: alternatives fibres for paper and paperboard production. AB - The pulping of Cynara cardunculus L. (cardoon) was performed under conditions for kraft, kraft-AQ and soda-AQ processes. The best results in terms of delignification degree, expressed as kappa number, pulp viscosity and screened yield, were obtained for the kraft-AQ process with 0.20% of anthraquinone (AQ). The papermaking potential of the selected pulp was studied attending to biometric fibre characterisation, refining aptitude, optical and strength properties. All properties were compared against a Eucalyptus globulus pulp at different refining degrees. The cardoon pulp was also evaluated concerning its potential to board manufacture, alone and in mixtures with pine pulp, giving rise to promising results for liner manufacture. PMID- 17398090 TI - Dual M3 antagonists-PDE4 inhibitors. Part 2: Synthesis and SAR of 3-substituted azetidinyl derivatives. AB - Introduction of 3-substituted azetidinyl substituents onto the 4,6 diaminopyrimidine scaffold allowed the improvement of PDE4 inhibiting activities. Preliminary in vivo activity in pulmonary inflammation models is reported. PMID- 17398091 TI - Antiproliferative activity of 4-chloro-5,6-dihydro-2H-pyrans. Part 2: Enhancement of drug cytotoxicity. AB - The Prins reaction was the basis to synthesize functionalized alkyl chlorodihydropyran derivatives. The inexpensive, stable, and environmentally friendly FeCl(3) promotes the cyclization. The method represents an efficient and regioselective manner to obtain in a single step chlorovinyl-TMS oxacycles. The in vitro antiproliferative activities of the compounds were examined in the human solid tumor cell lines A2780 (ovarian cancer), SW1573 (non-small cell lung cancer), and WiDr (colon cancer). Overall, the results show an enhancement in the cytotoxicity exhibited by the new analogs when compared to their parental compounds. PMID- 17398092 TI - The discovery of highly selective erbB2 (Her2) inhibitors for the treatment of cancer. AB - The synthesis and biological evaluation of potent and selective inhibitors of the erbB2 kinase is presented. Based on the 4-anilinoquinazoline chemotype, the syntheses of several new series of erbB2 inhibitors are described with quinazoline and pyrido[4,3-d]pyrimidine cores. The vast majority of these compounds are found to be >100x selective over the closely related EGFR kinase. Two lead compounds are further shown to have low clearance and moderate bioavailability in rat. PMID- 17398093 TI - Imidazole moiety replacements in the 3-(1H-benzo[d]imidazol-2-yl)pyridin-2(1H) one inhibitors of insulin-like growth factor receptor-1 (IGF-1R) to improve cytochrome P450 profile. AB - A series of 3-(1H-benzo[d]imidazol-2-yl)pyridin-2(1H)-one inhibitors of insulin like growth factor receptor-1 (IGF-1R) were examined in which the pendant imidazole moiety was replaced to improve selectivity for IGF-1R inhibition over cytochrome P450 (CYP). Synthesis and SAR of these compounds is presented. PMID- 17398094 TI - HPV infection and breast cancer. PMID- 17398095 TI - The Arl4 family of small G proteins can recruit the cytohesin Arf6 exchange factors to the plasma membrane. AB - The small GTPase Arf6 regulates endocytosis, actin dynamics, and cell adhesion, and one of its major activators is the exchange factor Arf nucleotide-binding site opener (ARNO), also called cytohesin-2 [1, 2]. ARNO must be recruited from the cytosol to the plasma membrane in order to activate Arf6, and in addition to a Sec7 nucleotide-exchange domain it contains a C-terminal pleckstrin homology (PH) domain that binds phosphoinositides [3, 4]. ARNO and its three relatives, cytohesin-1, Grp1/cytohesin-3, and cytohesin-4, are expressed as two splice variants, with either two or three glycines in a loop in the phosphoinositide binding pocket of the PH domain [5, 6]. The diglycine form binds PtdIns(3,4,5)P(3) with high affinity and mediates recruitment of cytohesins to the plasma membrane in response to insulin and growth factors [7, 8]. However, the triglycine form has only micromolar affinity for both PtdIns(3,4,5)P(3) and PtdIns(4,5)P(2), affinities that are insufficient to confer membrane recruitment, raising the question of how the triglycine forms of cytohesins are regulated [5, 9]. Here we show that three related Arf-like GTPases of unknown function, Arl4a, Arl4c, and Arl4d, are able to recruit ARNO and other cytohesins to the plasma membrane by binding to their PH domains irrespective of whether they are in the diglycine or triglycine form. The Arl4 family thus defines a signal-transduction pathway that can mediate the plasma-membrane recruitment of cytohesins independently of a requirement for the generation of PtdIns(3,4,5)P(3). PMID- 17398096 TI - Capicua regulates cell proliferation downstream of the receptor tyrosine kinase/ras signaling pathway. AB - Signaling via the receptor tyrosine kinase (RTK)/Ras pathway promotes tissue growth during organismal development and is increased in many cancers [1]. It is still not understood precisely how this pathway promotes cell growth (mass accumulation). In addition, the RTK/Ras pathway also functions in cell survival, cell-fate specification, terminal differentiation, and progression through mitosis [2-7]. An important question is how the same canonical pathway can elicit strikingly different responses in different cell types. Here, we show that the HMG-box protein Capicua (Cic) restricts cell growth in Drosophila imaginal discs, and its levels are, in turn, downregulated by Ras signaling. Moreover, unlike normal cells, the growth of cic mutant cells is undiminished in the complete absence of a Ras signal. In addition to a general role in growth regulation, the importance of cic in regulating cell-fate determination downstream of Ras appears to vary from tissue to tissue. In the developing eye, the analysis of cic mutants shows that the functions of Ras in regulating growth and cell-fate determination are separable. Thus, the DNA-binding protein Cic is a key downstream component in the pathway by which Ras regulates growth in imaginal discs. PMID- 17398097 TI - Regulation of Arf6 and ACAP1 signaling by the PTB-domain-containing adaptor protein GULP. AB - The GTPase Arf6 regulates multiple cellular processes, including endocytosis, secretion, phagocytosis, cell adhesion, and cell migration [1, 2]. The Arf6 specific GAP ACAP1 is a negative regulator of Arf6-mediated signaling [3-7]. However, regulation of ACAP1- and Arf6-mediated signaling by other cellular proteins is not well understood. GULP/CED-6 is a phosphotyrosine binding (PTB) domain-containing adaptor protein linked to engulfment of apoptotic cells [8-13] and to cholesterol homeostasis [14]. Here, we identify a novel role for GULP as a positive regulator of Arf6. Knockdown of GULP decreased cellular Arf6-GTP, whereas GULP overexpression increased cellular Arf6-GTP. At the mechanistic level, GULP influenced Arf6 at four levels. First, GULP bound directly to GDP bound Arf6 via its PTB domain. Second, GULP associated with the Arf6-GAP ACAP1 at endogenous levels. Third, GULP reversed the Arf6-GTP decrease induced by ACAP1, and countered the ACAP1-mediated inhibition of cell migration. Fourth, GULP, ACAP1, and GDP-bound Arf6 were part of a tripartite complex, suggesting sequestration of ACAP1 as one mechanism of GULP action. Taken together, these data identify GULP as a modifier of cellular Arf6-GTP through regulation of ACAP1. Because PTB-domain-containing adaptor proteins influence endocytosis and trafficking of membrane proteins and cell migration [15, 16], our data support a model wherein PTB-domain-containing adaptor proteins regulate Arf family proteins. PMID- 17398098 TI - Positive-feedback loops as a flexible biological module. AB - BACKGROUND: Bistability in genetic networks allows cells to remember past events and to make discrete decisions in response to graded signals. Bistable behavior can result from positive feedback, but feedback loops can have other roles in signal transduction as well. RESULTS: We introduced positive feedback into the budding-yeast pheromone response to convert it into a bistable system. In the presence of feedback, transient induction with high pheromone levels caused persistent pathway activation, whereas at lower levels a fraction of cells became persistently active but the rest inactivated completely. We also generated mutations that quantitatively tuned the basal and induced expression levels of the feedback promoter and showed that they qualitatively changed the behavior of the system. Finally, we developed a simple stochastic model of our positive feedback system and showed the agreement between our simulations and experimental results. CONCLUSIONS: The positive-feedback loop can display several different behaviors, including bistability, and can switch between them as a result of simple mutations. PMID- 17398099 TI - Dia-interacting protein modulates formin-mediated actin assembly at the cell cortex. AB - BACKGROUND: Mammalian Diaphanous (mDia)-related formins and the N-WASP-activated Arp2/3 complex initiate the assembly of filamentous actin. Dia-interacting protein (DIP) binds via its amino-terminal SH3 domain to the proline-rich formin homology 1 (FH1) domain of mDia1 and mDia2 and to the N-WASp proline-rich region. RESULTS: Here, we investigated an interaction between a conserved leucine-rich region (LRR) in DIP and the mDia FH2 domain that nucleates, processively elongates, and bundles actin filaments. DIP binding to mDia2 was regulated by the same Rho-GTPase-controlled autoinhibitory mechanism modulating formin-mediated actin assembly. DIP was previously shown to interact with and stimulate N-WASp dependent branched filament assembly via Arp2/3. Despite direct binding to both mDia1 and mDia2 FH2 domains, DIP LRR inhibited only mDia2-dependent filament assembly and bundling in vitro. DIP expression interfered with filopodia formation, consistent with a role for mDia2 in assembly of these structures. After filopodia retraction into the cell body, DIP expression induced excessive nonapoptotic membrane blebbing, a physiological process involved in both cytokinesis and amoeboid cell movement. DIP-induced blebbing was dependent on mDia2 but did not require the activities of either mDia1 or Arp2/3. CONCLUSIONS: These observations point to a pivotal role for DIP in the control of nonbranched and branched actin-filament assembly that is mediated by Diaphanous-related formins and activators of Arp2/3, respectively. The ability of DIP to trigger blebbing also suggests a role for mDia2 in the assembly of cortical actin necessary for maintaining plasma-membrane integrity. PMID- 17398100 TI - Interactions between Neisseria meningitidis and the complement system. AB - Meningococcal infection remains a worldwide health problem, and understanding the mechanisms by which Neisseria meningitidis evades host innate and acquired immunity is crucial. The complement system is vital for protecting individuals against N. meningitidis. However, this pathogen has evolved several mechanisms to avoid killing by human complement. Bacterial structures such as polysaccharide capsule and those which mimic or bind host molecules function to prevent complement-mediated lysis and phagocytosis. This review provides an update on the recent findings on the diverse mechanisms by which N. meningitidis avoids complement-mediated killing, and how polymorphisms in genes encoding human complement proteins affect susceptibility to this important human pathogen. PMID- 17398101 TI - Virus glycosylation: role in virulence and immune interactions. AB - The study of N-linked glycosylation as it relates to virus biology has become an area of intense interest in recent years due to its ability to impart various advantages to virus survival and virulence. HIV and influenza, two clear threats to human health, have been shown to rely on expression of specific oligosaccharides to evade detection by the host immune system. Additionally, other viruses such as Hendra, SARS-CoV, influenza, hepatitis and West Nile rely on N-linked glycosylation for crucial functions such as entry into host cells, proteolytic processing and protein trafficking. This review focuses on recent findings on the importance of glycosylation to viral virulence and immune evasion for several prominent human pathogens. PMID- 17398102 TI - Mercury in amalgam tattoos. PMID- 17398103 TI - Absorbed doses on patients undergoing tomographic exams for pre-surgery planning of dental implants. AB - The thermoluminescent (TL) dosimetry was used to measure entrance skin absorbed doses at anatomical points close to critical organs of patients undergoing tomographic techniques as part of a pre-surgery planning for dental implants. The dosimetric procedure was applied in 19 patients, and absorbed doses could be measured with a combined uncertainty down to 14%. Results showed that patient doses may be increased by a factor of 20 in the helical computed tomography compared to panoramic and spiral conventional tomographic exams. PMID- 17398104 TI - X-Ray energy dependence of the dose response of SIRAD radiation dosimeters. AB - SIRADs (self-indicating instant radiation alert dosimeters) are designed to measure accident radiation doses. As the energy of radiation is usually unknown in such situations, a detector with a weak energy dependence of its response to dose would be ideal. We have studied the energy dependence of the dose response of SIRADs in the range from 50kVp to 10MV, which corresponds to photon equivalent energies from 25.5keV to 2.2MeV. The response to the same dose at 25.5keV is (29+/-4)%(+/-1s) lower than the response at 1.4MeV. The response to a dose slowly increases with radiation energy. This energy dependence is relatively weak in comparison with the dependence for radiographic films and similar in magnitude to the dependence for lithium fluoride thermoluminescence dosimeters. This energy dependence of the response diminishes the accuracy of dose assessments in radiation fields of unknown energy, but does not significantly compromise the core ability of the devices to provide visual estimates of radiation doses. PMID- 17398105 TI - Changes in inflammatory processes associated with selective vulnerability following mild impairment of oxidative metabolism. AB - Abnormalities in oxidative metabolism and reductions of thiamine-dependent enzymes accompany many age-related neurodegenerative diseases. Thiamine deficiency (TD) produces a cascade of events including mild impairment of oxidative metabolism, activation of microglia, astrocytes and endothelial cells that leads to neuronal loss in select brain regions. The earliest changes occur in a small, well-defined brain region, the submedial thalamic nucleus (SmTN). In the present study, a micropunch technique was used to evaluate quantitatively the selective regional changes in mRNA and protein levels. To test whether this method can distinguish between changes in vulnerable and non-vulnerable regions, markers for neuronal loss (NeuN) and endothelial cells (eNOS) and inflammation (IL-1beta, IL-6 and TNF-alpha) in SmTN and cortex of control and TD mice were assessed. TD significantly reduced NeuN and increased CD11b, GFAP and ICAM-1 immunoreactivity in SmTN as revealed by immunocytochemistry. When assessed on samples obtained by the micropunch method, NeuN protein declined (-49%), while increased mRNA levels were observed for eNOS (3.7-fold), IL-1beta (43-fold), IL-6 (44-fold) and TNF-alpha (64-fold) in SmTN with TD. The only TD-induced change that occurred in cortex with TD was an increase in TNF-alpha (22-fold) mRNA levels. Immunocytochemical analysis revealed that IL-1beta, IL-6 and TNF-alpha protein levels increased in TD brains and colocalized with glial markers. The consistency of these quantitative results with immunocytochemical measurements validates the micropunch technique. The results demonstrate that TD induces quantitative, distinct inflammatory responses and oxidative stress in vulnerable and non-vulnerable regions that may underlie selective vulnerability. PMID- 17398106 TI - Valproic acid-mediated neuroprotection in intracerebral hemorrhage via histone deacetylase inhibition and transcriptional activation. AB - The modification of histone N-terminal tails by acetylation or deacetylation can alter the interaction between histones and DNA, and thus regulate gene expression. Recent experiments have demonstrated that valproic acid (VPA), a well known anti-epileptic drug, can directly inhibit histone deacetylase (HDAC) activity and cause the hyperacetylation of histones. Moreover, VPA has been shown to mediate neuronal protection by activating signal transduction pathways and by inhibiting proapoptotic factors. In this study, we attempted to determine whether VPA alleviates cerebral inflammation and perihematomal cell death after intracerebral hemorrhage (ICH). Adult male rats received intraperitoneal injections of 300 mg/kg VPA or PBS twice a day after ICH induction. VPA treatment inhibited hematoma expansion, perihematomal cell death, caspase activities, and inflammatory cell infiltration. In addition, VPA treatment had the following expressional effects; it activated the translations of acetylated histone H3, pERK, pAKT, pCREB, and HSP70; up-regulated bcl-2 and bcl-xl but down-regulated bax; and down-regulated the mRNAs of Fas-L, IL-6, MMP-9, MIP-1, MCP-1, and tPA. VPA-treated rats also showed better functional recovery from 1 day to 4 weeks after ICH. Here we show that VPA induces neuroprotection in a murine ICH model and that its neuroprotective effects are mediated by transcriptional activation following HDAC inhibition. PMID- 17398107 TI - Early cerebrovascular and parenchymal events following prenatal exposure to the putative neurotoxin methylazoxymethanol. AB - One of the most common causes of neurological disabilities are malformations of cortical development (MCD). A useful animal model of MCD consists of prenatal exposure to methylazoxymethanol (MAM), resulting in a postnatal phenotype characterized by cytological aberrations reminiscent of human MCD. Although postnatal effects of MAM are likely a consequence of prenatal events, little is known on how the developing brain reacts to MAM. General assumption is the effects of prenatally administered MAM are short lived (24 h) and neuroblast specific. MAM persisted for several days after exposure in utero in both maternal serum and fetal brain, but at levels lower than predicted by a neurotoxic action. MAM levels and time course were consistent with a different mechanism of indirect neuronal toxicity. The most prominent acute effects of MAM were cortical swelling associated with mild cortical disorganization and neurodegeneration occurring in absence of massive neuronal cell death. Delayed or aborted vasculogenesis was demonstrated by MAM's ability to hinder vessel formation. In vitro, MAM reduced synthesis and release of VEGF by endothelial cells. Decreased expression of VEGF, AQP1, and lectin-B was consistent with a vascular target in prenatal brain. The effects of MAM on cerebral blood vessels persisted postnatally, as indicated by capillary hypodensity in heterotopic areas of adult rat brain. In conclusion, these results show that MAM does not act only as a neurotoxin per se, but may additionally cause a short-lived toxic effect secondary to cerebrovascular dysfunction, possibly due to a direct anti-angiogenic effect of MAM itself. PMID- 17398108 TI - Cost reduction of perioperative coagulation management in cardiac surgery: value of "bedside" thrombelastography (ROTEM). AB - OBJECTIVE: Demographic changes and aggressive platelet inhibition have resulted in a marked increase in blood- and coagulation product expenditure and costs in cardiac surgery. We analyzed "bedside" coagulation test (ROTEM) in order to verify clot forming quality for the purpose of finding a cost-effective treatment path. METHODS: Annual treatment costs of all cardiosurgical patients were analyzed before (729 patients) and after (693 patients) implementation of "bedside" ROTEM. Cumulative numbers and costs of platelet concentrates (PltC), fresh frozen plasma (FFP), red blood cell units (RBC), and coagulation factors: pooled coagulation concentrates (PCC), recombinant factor VIIa (rFVIIa), factor XIII (FXIII), and fibrinogen were assessed. Average monthly numbers and costs were compared. Number of resternotomies and early mortality was assessed and compared in both periods. RESULTS: After ROTEM implementation cumulative RBC expenditure showed 25% decrease while PltC exhibited 50% decrease. FFP expenditure remained unchanged. PCC, FXIII were markedly reduced (-80%) while rFVIIa were entirely omitted. Fibrinogen, however, increased two-fold. Cumulative average monthly costs of all blood products decreased from 66,000 euro to 45,000 euro (-32%). Coagulation factor average monthly costs decreased from 60,000 euro to 30,000 euro (-50%) yielding combined savings of 44%. In contrast, average monthly costs for ROTEM were 1.580 euro. Total number of resternotomies decreased from 6.6% to 5.5% while early mortality (5.9%; 6.0%) remained stable. CONCLUSION: Cumulative costs for treatment of perioperative coagulation disorders can be reduced by "bedside" ROTEM analysis to achieve a selective substitution management. Saved costs for blood- and coagulation products clearly outweighed the expenses of ROTEM. Adequate differential coagulation management can therefore be cost-effective. PMID- 17398109 TI - Can preoperative modelling of individual neutrophil adhesion responses predict renal morbidity? AB - OBJECTIVE: Perioperative upregulation of the neutrophil adhesion molecule CD11b is associated with the development of renal impairment. We hypothesised that individual variation in neutrophil adhesion molecule responses to surgery influences renal outcomes and that this individual variability could be modelled prior to surgery and used to predict high risk patients. The developed model uses preoperative exposure of an individual patient's neutrophils to a fixed inflammatory stimulus and assessment of the basal and stimulated adhesion molecule CD11b expression. METHODS: Neutrophils were isolated from human volunteers undergoing cardiac surgery with cardiopulmonary bypass support. Basal and stimulated CD11b expression was measured using flow cytometry in preoperative neutrophil samples and compared to postoperative clinical performance. RESULTS: Patients with low levels of preoperative basal neutrophil CD11b expression had the greatest increase in CD11b following phorbol-12-myristate-13-acetate stimulation. This stimulated CD11b response correlated with changes in CD11b expression from preoperative to postoperative sampling. Preoperative basal CD11b expression showed a significant inverse relationship with postoperative creatinine levels. However, preoperative CD11b stimulation was not related to postoperative renal function. In addition preoperative basal CD11b expression correlated with adrenaline requirements and intra-aortic balloon pump usage. In contrast stimulated CD11b expression was significantly related to length of hospital stay and changes in the A-a gradient. CONCLUSIONS: Preoperative CD11b expression assessment might enable preoperative identification of patients who will mount an exaggerated and damaging neutrophil response to surgery which contributes to renal injury. Identification of these patients would then allow selective application of immunomodulatory therapies. PMID- 17398110 TI - Mid-term changes of left ventricular geometry and function after Dor, SAVE, and Overlapping procedures. AB - BACKGROUND: The aim of this study was to investigate the mid-term changes of left ventricular (LV) geometry and function after Dor, septal anterior ventricular exclusion (SAVE), and Overlapping ventricular remodeling procedures. METHODS: Forty-three patients who underwent LV reconstruction for end-stage ischemic heart disease, were divided into three groups, undergoing Dor (n=15), SAVE (n=12), and Overlapping procedures (n=16). Coronary artery bypass grafting and mitral annuloplasty were performed concomitantly when indicated. Left ventricular diastolic and systolic dimensions (LVDd and LVDs), LV end-diastolic and end systolic volume indexes (LVEDVI and LVESVI), LV ejection fraction (LVEF), deceleration time (DcT), sphericity index (SI), and grade of mitral regurgitation were measured on preoperative and postoperative (immediately and at intermediate term) echocardiography. RESULTS: In the Dor group, the LVEDVI and LVESVI were significantly reduced immediately after the operation, and increased again at intermediate follow-up. The SI was significantly increased immediately after the operation and increased thereafter in a linear fashion. In the SAVE group, the DcT was significantly reduced immediately after the operation and was not improved in the later stage. In the Overlapping group, the LVEDVI and LVESVI were significantly reduced, and remained as such at intermediate follow-up. The SI was not increased and remained almost unchanged after the operation. CONCLUSION: Progression in LV sphericity after the Dor procedure and persistent reduction of the DcT after the SAVE procedure seem to be procedure-related problems. The Overlapping procedure provided significant LV volume reduction, maintaining the most elliptical LV shape with acceptable early and late mortality. PMID- 17398111 TI - Preoperative red sudan administration to locate thoracic duct lesion in videothoracoscopy. PMID- 17398112 TI - Multivariate analysis of matrix-assisted laser desorption/ionization mass spectrometric data related to glycoxidation products of human globins in nephropathic patients. AB - To clarify the possible pathogenetic role of oxidation products originated from the glycation of proteins, human globins from nephropathic patients have been studied by matrix-assisted laser desorption/ionization mass spectrometry (MALDI), revealing not only unglycated and monoglycated globins, but also a series of different species. For the last ones, structural assignments were tentatively done on the basis of observed masses and expectations for the Maillard reaction pattern. Consequently, they must be considered only propositive, and the discussion which will follow must be considered in this view. In our opinion this approach does not seem to compromise the intended diagnostic use of the data because distinctions are valid even if the assignments are uncertain. We studied nine healthy subjects and 19 nephropathic patients and processed the data obtained from the MALDI spectra using a multivariate analysis. Our results showed that multivariate analytical techniques enable differential aspects of the profile of molecular species to be identified in the blood of end stage nephropathic patients. A correct grouping can be achieved by principal component analysis (PCA) and the results suggest that several products involved in carbonyl stress exist in nephropathic patients. These compounds may have a relevant role as specific markers of the pathological state. PMID- 17398113 TI - Synthetic surfaces as artificial antigen presenting cells in the study of T cell receptor triggering and immunological synapse formation. AB - T cell activation occurs when T cell receptors engage peptide-major histocompatibility complex (pMHC) molecules displayed on the surface of antigen presenting cells (APCs). Clustering of TCRs and other receptors in physical patterns at the T-APC interface forms a structure known as an immunological synapse (IS). Studies of the IS are challenging due to the cell-cell contact context of the governing interactions. Model surfaces as synthetic APCs have thus been developed, where the type, quantity, and physical arrangement of ligands displayed to T cells are precisely controlled. These model systems have provided important insights into the structure and function of the IS. PMID- 17398114 TI - Polyspecificity of T cell and B cell receptor recognition. AB - A recent workshop discussed the recognition of multiple distinct ligands by individual T cell and B cell receptors and the implications of this discovery for lymphocyte biology. The workshop recommends general use of the term polyspecificity because it emphasizes two fundamental aspects, the inherent specificity of receptor recognition and the ability to recognize multiple ligands. Many different examples of polyspecificity and the structural mechanisms were discussed, and the group concluded that polyspecificity is a general, inherent feature of TCR and antibody recognition. This review summarizes the relevance of polyspecificity for lymphocyte development, activation and disease processes. PMID- 17398115 TI - 3D imaging of microstructure of spruce wood. AB - Synchrotron radiation phase-contrast X-ray tomographic microscopy (srPCXTM) was applied to observation and identification of the features of spruce anatomy at the cellular lengthscale. The pilot experiments presented in the paper clearly revealed the features of the heartwood of Spruce (Picea abies [L.] Karst.), such as lumina and pits connecting the lumina, with a theoretical voxel size of 0.7 x 0.7 x 0.7 microm(3). The experiments were carried out on microspecimens of heartwood, measuring approximately 200 by 200 micrometers in cross-section. The technique for production and preparation of wood microsamples was developed within the framework of this investigation. The total porosity of the samples was derived and the values of the microstructural parameters, such as the diameters of tracheid, cell wall thicknesses and pit diameters were assessed non invasively. Microstructural features as thin/small as approximately 1.5 microm were revealed and reconstructed in 3D. It is suggested that the position of sub voxel-sized features (such as position of tori in the bordered pit pairs) can be determined indirectly using watershed segmentation. Moreover, the paper discusses the practical issues connected with a pipelined phase-contrast synchrotron-based microtomography experiment and the possible future potentials of this technique in the domain of wood science. PMID- 17398116 TI - The cognitive and neural correlates of "tactile consciousness": a multisensory perspective. AB - People's awareness of tactile stimuli has been investigated in far less detail than their awareness of stimuli in other sensory modalities. In an attempt to fill this gap, we provide an overview of studies that are pertinent to the topic of tactile consciousness. We discuss the results of research that has investigated phenomena such as "change blindness", phantom limb sensations, and numerosity judgments in tactile perception, together with the results obtained from the study of patients affected by deficits that can adversely affect tactile perception such as neglect, extinction, and numbsense. The similarities as well as some of the important differences that have emerged when visual and tactile conscious information processing have been compared using similar experimental procedures are highlighted. We suggest that conscious information processing in the tactile modality cannot be separated completely from the more general processing of spatial information in the brain. Finally, the importance of considering tactile consciousness within the larger framework of multisensory information processing is also discussed. PMID- 17398117 TI - fMRI reveals a preference for near viewing in the human parieto-occipital cortex. AB - Posterior parietal cortex in primates contains several functional areas associated with visual control of body effectors (e.g., arm, hand and head) which contain neurons tuned to specific depth ranges appropriate for the effector. For example, the macaque ventral intraparietal area (VIP) is involved in head movements and is selective for motion in near-space around the head. We used functional magnetic resonance imaging to examine activation in the putative human VIP homologue (pVIP), as well as parietal and occipital cortex, as a function of viewing distance when multiple cues to target depth were available (Expt 1) and when only oculomotor cues were available (Expt 2). In Experiment 1, subjects viewed stationary or moving disks presented at three distances (with equal retinal sizes). Although activation in pVIP showed no preference for any particular spatial range, the dorsal parieto-occipital sulcus (dPOS) demonstrated a near-space preference, with activation highest for near viewing, moderate for arm's length viewing, and lowest for far viewing. In Experiment 2, we investigated whether the near response alone (convergence of the eyes, accommodation of the lens and pupillary constriction) was sufficient to elicit this same activation pattern. Subjects fixated lights presented at three distances which were illuminated singly (with luminance and visual angle equated across distances). dPOS displayed the same gradient of activation (Near>Medium>Far) as that seen in Experiment 1, even with reduced cues to depth. dPOS seems to reflect the status of the near response (perhaps driven largely by vergence angle) and may provide areas in the dorsal visual stream with spatial information useful for guiding actions toward targets in depth. PMID- 17398118 TI - Discordant white matter N-acetylasparate and diffusion MRI measures suggest that chronic metabolic dysfunction contributes to axonal pathology in multiple sclerosis. AB - Diffusion MRI and magnetic resonance spectroscopic measurements of selectively neuronally localised N-acetylaspartate (NAA) both have been used widely to assess white matter integrity and axonal loss. We have tested directly the relationship between changes in diffusion MRI parameters and NAA concentrations in the corpus callosum (CC) in an in vivo study of patients with MS. Fifteen MS patients (median EDSS 2.5, range 1-4) were studied with T(1) anatomical, T(2)-weighted, and diffusion-sensitised MRI and PRESS single-voxel MRS. A recently described method, tract-based spatial statistics (TBSS) [Smith, S.M., Jenkinson, M., Johansen-Berg, H., Rueckert, D., Nichols, T.E., Mackay, C.E. et al., 2006. Tract based spatial statistics: voxelwise analysis of multi-subject diffusion data. Neuroimage 31, 1487-1505] also was used to perform exploratory voxelwise whole brain analysis of white matter diffusion fractional anisotropy (FA). We found a strong correlation between callosal size and both mean FA (r=0.68, p<0.005) (related specifically to changes in the radial tensor component) and mean inter hemispheric motor tract connectivity probability (r=0.74, p<0.001). TBSS confirmed that the diffusion anisotropies of white matter voxels specifically within the callosum were correlated with the callosal size. Individual patient global T(2) lesion volumes were correlated with both the probability of callosal connectivity (r=-0.69, p<0.005) and fractional anisotropy across the callosum (r= 0.76, p<0.001). However, absolute concentrations of NAA from the voxel showed no correlation with callosal cross-sectional area, mean connectivity or fractional anisotropy within the callosal pathway. We conclude that diffusion MRI shows changes consistent with sensitivity to axonal loss, but that relative NAA changes are not necessarily related directly to this. Axonal metabolic function, independent of structural integrity, may be a major determinant of NAA measures in MS. PMID- 17398119 TI - Brain activation mediates the association between structural abnormality and symptom severity in schizophrenia. AB - Thought disorder is a symptom of schizophrenia expressed as disorganized or incoherent speech. Severity of thought disorder correlates with decreased left superior temporal gyrus grey matter volume and cortical activation in posterior temporal regions during the performance of language tasks. The goal of this study was to determine whether language-related activation mediates the association between thought disorder and left superior temporal lobe grey matter volume. 12 patients with schizophrenia were assessed for thought disorder. FMRI images were acquired for each subject while they listened to English speech, along with a high resolution structural image. Thought disorder was used as a covariate in the functional analysis to identify brain regions within which activation correlated with symptom severity. Voxel based morphometry was used to calculate grey matter volume of the planum temporale. A mediation model waste-tested using a four-step multiple regression approach incorporating cortical volume, functional activation and symptom severity. Thought disorder correlated with activation in a single cluster within the left posterior middle temporal gyrus during listening to speech. Grey matter volume within the planum temporale correlated significantly with severity of thought disorder and activation within the functional cluster. Regressing thought disorder on grey matter volume and BOLD response simultaneously led to a significant reduction in the correlation between grey matter volume and thought disorder. These results support the hypothesis that the association between decreased grey matter volume in the left planum temporale and severity of thought disorder is mediated by activation in the posterior temporal lobe during language processing. PMID- 17398120 TI - A systematic comparison of kinetic modelling methods generating parametric maps for [(11)C]-(R)-PK11195. AB - [(11)C]-(R)-PK11195 is presently the most widely used radiotracer for the monitoring of microglia activity in the central nervous system (CNS). Microglia, the resident immune cells of the brain, play a critical role in acute and chronic diseases of the central nervous system and in host defence against neoplasia. The purpose of this investigation was to evaluate the reliability and sensitivity of five kinetic modelling methods for the formation of parametric maps from dynamic [(11)C]-(R)-PK11195 studies. The methods we tested were the simplified reference tissue model (SRTM), basis pursuit, a simple target-to-reference ratio, the Logan plot and a wavelet based Logan plot. For the reliability assessment, the test retest data consisted of four Alzheimer's patients that were scanned twice at approximately a six-week interval. For the sensitivity assessment, comparison of [(11)C]-(R)-PK11195 binding in Huntington's disease (HD) patients and normal subjects was performed using a group contrast to localize significant increases in mean pixel volume of distribution (VD) in HD. In all instances, a reference region kinetic extracted by a supervised clustering technique was used as input function. Reliability was assessed by use of the intra-class correlation coefficient (ICC) across a wide set of anatomical regions and it was found that the wavelet-based Logan plot, basis pursuit and SRTM gave the highest ICC values on average. The same methods produced the highest z-scores resulting from increases in mean striatal VD in HD patients compared with controls. The reference-to-target ratio and the Logan graphical approach were significantly less reliable and less sensitive. PMID- 17398121 TI - The disunity of "Mysidacea" (Crustacea). AB - New studies on malacostracan relationships have drawn attention to issues concerning monophyly of the order Mysidacea, manifested in recent crustacean classifications that treat the taxon as two separate orders, Lophogastrida and Mysida. We present molecular phylogenies of these orders based on complete sequences of nuclear small-subunit ribosomal DNA (18S rRNA), and morphological evidence is used to revise the classification of the order Mysida to better reflect evolutionary history. A secondary structure model for 18S rRNA was constructed and used to assign putative stem and loop regions to two groups of partitions for phylogenetic analyses. Phylogenies were estimated by maximum likelihood, Bayesian inference, and maximum-parsimony. The analyses gave strong support for three independently derived lineages, represented by three monophyletic groups, Lophogastrida, Stygiomysida, and Mysida. The family Petalophthalmidae is considered as sister group to the family Mysidae, and Boreomysinae and Rhopalophthalminae are the most early derived of the Mysidae. The tribes contained in the current classification of the subfamily Mysinae are not well-supported by either molecular data or morphology. PMID- 17398122 TI - Efficacy and safety of opioids for osteoarthritis: a meta-analysis of randomized controlled trials. AB - OBJECTIVES: To determine the analgesic effectiveness, the effect on physical function and the safety of opioids in patients with osteoarthritis (OA). SEARCH STRATEGY: A systematic literature search was performed in electronic databases up to October 2006. A hand search of references was also performed. SELECTION CRITERIA: All randomized controlled trials evaluating the efficacy and/or the safety of opioids vs placebo or non-opioid analgesics in patients with OA were selected. DATA COLLECTION AND ANALYSIS: Data were collected using a predetermined form. Statistical analysis determined in each trial the effect size to assess the magnitude of treatment effect and the number needed to harm (NNH) to evaluate opioids safety. MAIN RESULTS: Eighteen randomized placebo-controlled trials were analyzed, i.e., a total of 3244 participants who received opioids and 1612 who received placebo. The mean trial duration was 13+/-18 weeks. The pooled effect sizes of all opioids vs placebo for pain intensity and physical function were 0.79 (95% confidence interval, CI, -0.98 to -0.59) and -0.31 (95% CI -0.39 to 0.24), respectively. The NNH was calculated to be 5 vs placebo. The number of studies (n=4) that compared opioids with non-opioid analgesics (paracetamol and non-steroidal anti-inflammatory drugs) was too limited to provide robust data. CONCLUSIONS: Opioids significantly decrease pain intensity and have small benefits on function compared with placebo in patients with OA. Adverse events, although reversible and not life threatening, often cause participants to stop taking the medication and could limit opioid usefulness. Moreover, the long-term efficacy and safety of these drugs for OA is yet to be determined due to the short mean trial duration. PMID- 17398123 TI - The Toll-interleukin-1 receptor member SIGIRR regulates colonic epithelial homeostasis, inflammation, and tumorigenesis. AB - Despite constant contact with the large population of commensal bacteria, the colonic mucosa is normally hyporesponsive to these potentially proinflammatory signals. Here we report that the single immunoglobulin IL-1 receptor-related molecule (SIGIRR), a negative regulator for Toll-IL-1R signaling, plays a critical role in gut homeostasis, intestinal inflammation, and colitis-associated tumorigenesis by maintaining the microbial tolerance of the colonic epithelium. SIGIRR-deficient (Sigirr(-/-)) colonic epithelial cells displayed commensal bacteria-dependent homeostatic defects, as shown by constitutive upregulation of inflammatory genes, increased inflammatory responses to dextran sulfate sodium (DSS) challenge, and increased Azoxymethane (AOM)+DSS-induced colitis-associated tumorigenesis. Gut epithelium-specific expression of the SIGIRR transgene in the SIGIRR-deficient background reduced the cell survival of the SIGIRR-deficient colon epithelium, abrogated the hypersensitivity of the Sigirr(-/-) mice to DSS induced colitis, and reduced AOM+DSS-induced tumorigenesis. Taken together, our results indicate that epithelium-derived SIGIRR is critical in controlling the homeostasis and innate immune responses of the colon to enteric microflora. PMID- 17398125 TI - Why cells move messages: the biological functions of mRNA localization. AB - RNA localization is a widespread mechanism that allows cells to spatially control protein function by determining their sites of synthesis. In embryos, localized mRNAs are involved in morphogen gradient formation or the asymmetric distribution of cell fate determinants. In somatic cell types, mRNA localization contributes to local assembly of protein complexes or facilitates protein targeting to organelles. Long-distance transport of specific mRNAs in plants allows coordination of developmental processes between different plant organs. In this review, we will discuss the biological significance of different patterns of mRNA localization. PMID- 17398124 TI - Dendritic cells prime natural killer cells by trans-presenting interleukin 15. AB - Natural killer (NK) cells are important effector cells in the control of infections. The cellular and molecular signals required for NK cell activation in vivo remain poorly defined. By using a mouse model for the inducible ablation of dendritic cells (DCs), we showed that the in vivo priming of NK cell responses to viral and bacterial pathogens required the presence of CD11c(high) DCs. After peripheral Toll-like receptor (TLR) stimulation, NK cells were recruited to local lymph nodes, and their interaction with DCs resulted in the emergence of effector NK cells in the periphery. NK cell priming was dependent on the recognition of type I IFN signals by DCs and the subsequent production and trans-presentation of IL-15 by DCs to resting NK cells. CD11c(high) DC-derived IL-15 was necessary and sufficient for the priming of NK cells. Our data define a unique in vivo role of DCs for the priming of NK cells, revealing a striking and previously unappreciated homology to T lymphocytes of the adaptive immune system. PMID- 17398126 TI - Temperature dependent properties of a kinesin-3 motor protein from Thermomyces lanuginosus. AB - Kinesins are cytoskeletal motor proteins that share a common mechanochemical motor domain, and are responsible for trafficking macromolecules. Here we report the cloning and characterization of a monomeric, kinesin-3 (TKIN) from Thermomyces lanuginosus. TKIN displayed a maximum rate of ATP hydrolysis at approximately 55 degrees C; the K(m)(ATP) was also significantly greater at 50 degrees C. Gliding motility rates reached a maximum of 5.5 microms(-1) at 45 degrees C, which is among the highest rates reported for kinesin. Arrhenius energy barriers were calculated to be approximately 103 kJmol(-1), nearly twofold greater than other mesophilic kinesin motors. The enthalpy of activation and entropy activation of TKIN were also significantly greater when compared to other mesophilic kinesins. A thermally induced aggregation of TKIN, which could be moderated by the addition of ATP, was observed at temperatures above 45 degrees C. Together, these results illustrate the kinetic response and stability of this unique motor protein at elevated temperatures. PMID- 17398127 TI - Telomeres and telomerase: Biological and clinical importance in dogs. AB - In recent years in human oncology the enzyme telomerase has emerged as an ideal target for cancer therapy. This has led to the assessment of telomerase in cancers in companion animals, mainly dogs and these studies confirm that in dogs, like humans, telomere maintenance by telomerase is the primary mechanism by which cancer cells overcome their mortality and extend their lifespan. This review aims to provide an introduction to the biology of telomeres and telomerase and to discuss some of the telomere/telomerase directed therapeutic methodologies currently under development which may be of benefit to the canine cancer patient. PMID- 17398128 TI - Conceptus-related measurements during the first trimester of bovine pregnancy. AB - In order to determine the variability inherent in conceptus-related measurements in first trimester bovine pregnancies, conceptus and fetometric parameters from beef cattle pregnancies (n=103) estimated to be between Days 36 and 103 of gestation were examined. During this period, the protein concentration of amniotic fluid ranged between 0.181 and 0.501mg/mL. The amniotic fluid volume gradually increased from <1mL at Day 36 to 950mL at Day 103 (R(2)=0.9275) and amniotic compartment dimensions (length, R(2)=0.9713; width, R(2)=0.9802) increased predictably with fetal growth. Conversely, allantoic fluid protein concentration and volume correlated weakly with fetal age. A significant linear correlation existed between fetal crown rump length (CRL) and crown nose length (R(2)=0.9899) confirming that either measurement can be employed in the ultrasonographic estimation of fetal age. The amniotic compartment and fetometric data presented here have both research and clinical value, particularly in relation to fetal development evaluation and pregnancy viability diagnosis. PMID- 17398129 TI - Allometric dependence of the life span of mammal erythrocytes on thermal stability and sphingomyelin content of plasma membranes. AB - Thermal stability of erythrocyte membrane is a measure for its ability to maintain permeability barrier at deleterious conditions. Hence, it could impact the resistance of erythrocytes against detrimental factors in circulation. In this study the thermostability of erythrocyte membranes was expressed by the temperature, T(go), at which the transmembrane gradient of ion concentration rapidly dissipated during transient heating. T(go) is the inducing temperature of the membrane transition that activated passive ion permeability at hyperthermia causing thermal hemolysis. A good allometric correlation of T(go) to the resistance against thermal hemolysis and the life span of erythrocytes were found for 13 mammals; sheep, cow, goat, dog, horse, man, rabbit, pig, cat, hamster, guinea pig, rat, and mouse. For the same group, the values of T(go) were strictly related to the sphingomyelin content of erythrocyte membranes. The residual ion permeability, P, was temperature activated from 38 to 57 degrees C with activation energy of 250+/-15 kJ/mol that strongly differed from that below 37 degrees C. The projected value of P at 37 degrees C was about half that of residual physiological permeability for Na+ and K+ that build ground for possible explanation of the life span vs membrane thermostability allometric correlation. PMID- 17398130 TI - Osmoregulatory role of the brood pouch in the euryhaline Gulf pipefish, Syngnathus scovelli. AB - Many species of pipefish exhibit a reversal of parental roles, in which females insert eggs into the brood pouch of the male where they are incubated until the end of embryonic development. While the significance of the male brood pouch has been examined for over a century, the role of the pouch is still unclear. One possible function is to aid in osmoregulation by buffering embryos from the external environment. To investigate this role, the euryhaline Gulf pipefish, Syngnathus scovelli, was collected and maintained in either a low salinity or a saltwater environment. Changes in plasma and pouch fluid osmolality and morphological changes of the pouch were examined. Brood pouch fluid was similar to male plasma during the early and late stages of the brooding period for low salinity males, but was significantly hyperosmotic during the middle of the brooding period. In saltwater males, brood pouch fluid was similar to plasma during early brooding, but became hyperosmotic as brood time progressed. The brood pouch epithelium of both low salinity and saltwater males contained mitochondria-rich cells. In early brooding saltwater males these cells contained an apical opening into the pouch lumen. Osmotic and morphological differences observed suggest that the brood pouch plays an active role in regulating osmotic concentration of the pouch fluid. Additionally, pouch fluid concentration may be regulated more during early stages of embryonic development. PMID- 17398131 TI - Abscess of the corpus cavernosum. PMID- 17398132 TI - Detection and identification of bacterial enteropathogens by polymerase chain reaction and conventional techniques in childhood acute gastroenteritis in Gaza, Palestine. AB - BACKGROUND: Acute gastroenteritis and diarrhea are common and costly problems that cause significant morbidity and mortality in children worldwide. In Palestine, diarrhea is one of the major causes of outpatient visits and hospitalizations. METHODS: To improve knowledge on the etiology of gastroenteritis and diarrhea in our patient population, stool specimens from 150 children under 5 years of age suffering from acute gastroenteritis were investigated for various common bacterial enteropathogens by conventional and molecular techniques. RESULTS: Bacterial enteropathogens were detected in 17.3% of the diarrheal samples. Shigella spp was the most common bacterial pathogen (6.0%), followed by Campylobacter coli/jejuni (4.7%), Escherichia coli O157:H7 (4.7%), and Salmonella spp (2.0%). Shigella and Salmonella isolates were tested for their susceptibility to common antimicrobial agents and most of the Shigella isolates were resistant to ampicillin, trimethoprim/sulfamethoxazole and doxycycline and most of the Salmonella isolates showed resistant to ampicillin, trimethoprim/sulfamethoxazole, doxycycline and nalidixic acid. CONCLUSIONS: The results highlight the value of using a combination of traditional and molecular techniques (PCR) in the diagnosis of bacterial gastroenteritis. Furthermore, this study demonstrated that E. coli O157:H7 and Campylobacter, which are not screened for routinely in the Gaza Strip, were significant enteropathogens. PMID- 17398133 TI - The naturally acquired immunity in severe malaria and its implication for a PfEMP 1 based vaccine. AB - Malaria vaccine development has so far been largely focused on antigens involved in parasite invasion pathways rather than on antigens associated with severe disease and naturally acquired immunity. Individuals repeatedly exposed to Plasmodium falciparum will eventually become immune to severe disease. Parasite derived antigens expressed on the infected red blood cell (iRBC) surface are the main targets of protective immunity and can be explored as a rational alternative in development of an anti-malaria vaccine. PMID- 17398134 TI - CTLA-4 blockade differentially influences the outcome of non-lethal and lethal Plasmodium yoelii infections. AB - An immune response against malaria has to be tightly controlled. The production of pro-inflammatory cytokines is required to control parasites but the same cytokines are also involved in severe malaria. We have shown that CTLA-4 expression during Plasmodium berghei malaria dampens the immune response. This strain provokes a pro-inflammatory immune response that is associated with the pathology of cerebral malaria. Accordingly a blockade of CTLA-4 during the blood stage of P. berghei malaria leads to an exacerbation of disease. To analyze the effects of a CTLA-4 blockade in a malaria model which is not prone to immune pathology we employed P. yoelii infection. Blood-stage infection led to a rapid induction of CTLA-4 on T cells. Using the non-lethal P. yoelii strain Py17NL we found that a blockade of CTLA-4 resulted in an increased T cell activation and IFN-gamma production, which was accompanied by a lower peak parasitemia and earlier parasite clearance. In contrast, blockade of CTLA-4 during infection with a P. yoelii strain exhibiting a higher parasitemia induced markedly increased serum-levels of TNF-alpha, which was associated with severe inflammation and reduced survival. PMID- 17398135 TI - Peptide based malaria vaccine development: personal considerations. AB - A brief historical account of synthetic peptide approach to malaria vaccine discovery and development, and recent accomplishments is presented. Application of this approach to high-throughput antigen discovery using the genome information is described. The need of a truly global effort and cooperation to defeat malaria is also discussed. PMID- 17398137 TI - Development of vaccines against Plasmodium falciparum malaria: taking lessons from naturally acquired protective immunity. AB - The acquisition of substantial anti-malarial protection in people naturally exposed to P. falciparum is often cited as evidence that malaria vaccines can be developed, but is rarely used to guide the development. We are pursuing the development of vaccines based on antigens and immune responses that appear key in naturally acquired protection. PMID- 17398139 TI - [Against the obligation of single embryo transfer]. PMID- 17398140 TI - Do 'sliders' slide and 'tensioners' tension? An analysis of neurodynamic techniques and considerations regarding their application. AB - Despite the high prevalence of carpal tunnel syndrome and cubital tunnel syndrome, the quality of clinical practice guidelines is poor and non-invasive treatment modalities are often poorly documented. The aim of this cadaveric biomechanical study was to measure longitudinal excursion and strain in the median and ulnar nerve at the wrist and proximal to the elbow during different types of nerve gliding exercises. The results confirmed the clinical assumption that 'sliding techniques' result in a substantially larger excursion of the nerve than 'tensioning techniques' (e.g., median nerve at the wrist: 12.6 versus 6.1mm, ulnar nerve at the elbow: 8.3 versus 3.8mm), and that this larger excursion is associated with a much smaller change in strain (e.g., median nerve at the wrist: 0.8% (sliding) versus 6.8% (tensioning)). The findings demonstrate that different types of nerve gliding exercises have largely different mechanical effects on the peripheral nervous system. Hence different types of techniques should not be regarded as part of a homogenous group of exercises as they may influence neuropathological processes differently. The findings of this study and a discussion of possible beneficial effects of nerve gliding exercises on neuropathological processes may assist the clinician in selecting more appropriate nerve gliding exercises in the conservative and post-operative management of common neuropathies. PMID- 17398142 TI - Formation of micronuclei in erythrocytes of the fathead minnow (Pimephales promelas) after acute treatment with mitomycin C or cyclophosphamide. AB - Despite the widespread use of the fathead minnow in ecotoxicology, there have been relatively few studies on genotoxicity biomarkers in this small, warm-water fish species. Consequently, we investigated the effect of two known genotoxins, mitomycin C and cyclophosphamide, on micronucleus induction in spleen and peripheral blood erythrocytes of this species. Initially, 96-h experiments after intra-peritoneal (i.p.) injections of mitomycin C and cyclophosphamide were undertaken to determine the maximum tolerated dose (MTD). From these studies, MTDs of 10 and 400 mg/kg, respectively, were obtained: doses that were higher than those reported for other fish species. Next, an assessment of micronucleus induction at 1, 2, 4, 8 and 14 days after injection was undertaken for each compound at the MTD. Mitomycin C at 10 mg/kg significantly induced micronuclei in erythrocytes from the spleen, but not from the peripheral blood, at 8 and 14 days. In addition, the overall levels of micronuclei observed were lower than most previously published data from other fish species. In contrast to mitomycin C, treatment with 400 mg/kg cyclophosphamide failed to significantly induce micronuclei in erythrocytes from any of the tissues employed, in contrast to previous reports of significant induction in other species. The reasons for the apparent relative insensitivity of the fathead minnow to these clastogens, with respect to both MTDs and micronucleus induction, are not clear. The fathead minnow, however, has previously been described as relatively insensitive compared to other fish species with respect to selected carcinogens and cytochrome P450 inducers; the latter suggesting that the lack of a significant induction following cyclophosphamide exposure may be due to low metabolic activation in vivo. Consequently, further clarifying work is required to delineate the response shown, considering the extensive use of this species in ecotoxicology research and regulatory testing. PMID- 17398143 TI - Raman spectra of organic compounds kladnoite (C6H4(CO)2NH) and hoelite (C14H8O2)- rare sublimation products crystallising on self-ignited coal heaps. AB - As minerals, aromatic compounds occur very rarely in nature. Not more than 10 of such minerals are known and most of them were described in the coal deposits where they were formed as a result of coal bed fires or burning of coal waste heaps. Raman spectra of kladnoite C(6)H(4)(CO)(2)NH (natural phthalimide) and hoelite C(14)H(8)O(2), (natural 9,10-anthraquinone) display complex features. Raman microspectroscopy help to detect these phases non-destructively directly in the frame of rare samples. Investigated minerals are transformation products formed from gaseous phase originating in natural pyrolytical process occurring in the frame of coal heaps and coal series outcrops. It is recommended to include Raman spectroscopic characteristics of similar materials in databases for exobiological studies. PMID- 17398144 TI - Perchlorate mixed-ligand copper(II) complexes of beta-diketone and ethylene diamine derivatives: thermal, spectroscopic and biochemical studies. AB - The present work carried out a study on perchlorate mixed-ligand copper(II) complexes which have been synthesized from ethylenediamine derivatives (3a-c) and beta-diketones. These complexes, namely [Cu(DA-Cl)(acac)H(2)O]ClO(4)4, [Cu(DA Cl)(bzac)H(2)O]H(2)O.ClO(4)5, [Cu(DA-OMe)(acac)H(2)O]ClO(4)6, [Cu(DA OMe)(bzac)H(2)O]ClO(4)7, [Cu(DA-H)(acac)H(2)O]2H(2)O.ClO(4)8 and [Cu(DA H)(bzac)H(2)O]ClO(4)9 (where acac, acetylacetonate and bzac, benzoylacetonate) were characterized by elemental analysis, spectral (IR and UV-vis) and magnetic moment measurements. Thermal properties and decomposition kinetics of all complexes are investigated. The interpretation, mathematical analysis and evaluation of kinetic parameters (E, A, DeltaH, DeltaS and DeltaG) of all thermal decomposition stages have been evaluated using Coats-Redfern equation. The biochemical studies showed that, the diamines 3a-c have powerful effects on degradation of DNA and protein. The antibacterial screening demonstrated that, the diamine (DA-Cl), 3b has the maximum and broad activities against Gram +ve and Gram -ve bacterial strains. PMID- 17398145 TI - An effective computerized reminder for contact isolation of patients colonized or infected with resistant organisms. AB - PURPOSE: To improve contact isolation rates among patients admitted to the hospital with a known history of infection with Methicillin-resistant Staphylococcus aureus (MRSA) and Vancomycin-resistant Enterococci (VRE). METHODS: A before and after interventional study implementing computerized reminders for contact isolation between February 25, 2005 and February 28, 2006. We measured rates of appropriate contact isolation, and time to isolation for the 4 month pre intervention period, and the 12 month intervention period. We conducted a survey of ordering physicians at the midpoint of the intervention period. RESULTS: Implementing a computerized reminder increased the rate of patients appropriately isolated from 33% to fully 89% (P<0.0001). The median time to writing contact isolation orders decreased from 16.6 to 0.0 h (P<0.0001). Physicians accepted the order 80% of the time on the first or second presentation. Ninety-five percent of physicians felt the reminder had no impact on workflow, or saved them time. CONCLUSION: A human reviewed computerized reminder can achieve high rates of compliance with infection control recommendations for contact isolation, and dramatically reduce the time to orders being written upon admission. PMID- 17398146 TI - Combination of EEG and ECG for improved automatic neonatal seizure detection. AB - OBJECTIVE: Neonatal seizures are the most common central nervous system disorder in newborn infants. A system that could automatically detect the presence of seizures in neonates would be a significant advance facilitating timely medical intervention. METHODS: A novel method is proposed for the robust detection of neonatal seizures through the combination of simultaneously-recorded electroencephalogram (EEG) and electrocardiogram (ECG). A patient-specific and a patient-independent system are considered, employing statistical classifier models. RESULTS: Results for the signals combined are compared to results for each signal individually. For the patient-specific system, 617 of 633 (97.52%) expert-labelled seizures were correctly detected with a false detection rate of 13.18%. For the patient-independent system, 516 of 633 (81.44%) expert-labelled seizures were correctly detected with a false detection rate of 28.57%. CONCLUSIONS: A novel algorithm for neonatal seizure detection is proposed. The combination of an ECG-based classifier system with a novel multi-channel EEG based classifier system has led to improved seizure detection performance. The algorithm was evaluated using a large data-set containing ECG and multi-channel EEG of realistic duration and quality. SIGNIFICANCE: Analysis of simultaneously recorded EEG and ECG represents a new approach in seizure detection research and the detection performance of the proposed system is a significant improvement on previous reported results for automated neonatal seizure detection. PMID- 17398147 TI - The error negativity in nonmedicated and medicated patients with Parkinson's disease. AB - OBJECTIVE: It has been hypothesized that the error negativity (Ne or ERN) is modulated by the midbrain dopaminergic system. Thus, in a depleted dopaminergic system as seen in patients with Parkinson's disease (PD) one would expect an attenuated Ne. However, studies investigating the error negativities in medicated patients with PD have produced contradictory results and the present study was designed to explore this relationship further. METHODS: Using the event-related potential technique and an Eriksen flanker paradigm, we examined error negativities in nonmedicated (drug naive) and medicated PD patients and compared them to those of healthy controls. RESULTS: (a) The error negativities of the nonmedicated and medicated PD patients were attenuated compared to those of healthy elderly controls at frontocentral scalp sites; and (b) nonmedicated and medicated PD patients produced error negativities similar to each other. CONCLUSIONS: PD results in diminished error negativities both in the early stage nonmedicated patients and in the later stage medicated patients. SIGNIFICANCE: Because both patient groups have reduced dopaminergic functioning compared to healthy controls, these findings are consistent with Ne amplitude being sensitive to modulations in that system. PMID- 17398148 TI - Direct demonstration of the effects of repetitive paired-pulse transcranial magnetic stimulation at I-wave periodicity. AB - OBJECTIVE: To investigate the central nervous system level at which paired-pulse repetitive transcranial magnetic stimulation at I-wave periodicity (iTMS) produces a facilitation of motor evoked potential (MEP) amplitude. METHODS: In one conscious patient who had an electrode implanted in the cervical epidural space for the control of pain, we recorded corticospinal volleys evoked before, during and after iTMS of the motor cortex. Moreover, we compared MEPs to TMS and cervico-medullary junction stimulation before and after iTMS in a separate group of five healthy subjects. RESULTS: In the patient with the epidural electrode, during iTMS there was progressive increase of MEP amplitude, and by the end of the intervention period MEP increased by more than 300%. The pronounced increase in MEP amplitude was paralleled by a slight increase in the amplitude of epidural volleys. An increased MEP amplitude (more than 200%) was still evident 3 min after the end of iTMS. In the five healthy subjects, iTMS produced a facilitation of MEPs evoked by transcranial magnetic stimulation but had no effect on CMEPs evoked by cervico-medullary junction stimulation. CONCLUSIONS: The results indicate that iTMS leads to an increase in corticomotor excitability at a supraspinal level, and that this may include circuits in addition to those involved in I-wave generation. SIGNIFICANCE: iTMS increases cortical excitability more widely than the I-wave networks that it targets. PMID- 17398149 TI - Analysis of the generators of epileptic activity in early-onset childhood benign occipital lobe epilepsy. AB - OBJECTIVE: The Panayiotopoulos type of idiopathic occipital epilepsy has peculiar and easily recognizable ictal symptoms, which are associated with complex and variable spike activity over the posterior scalp areas. These characteristics of spikes have prevented localization of the particular brain regions originating clinical manifestations. We studied spike activity in this epilepsy to determine their brain generators. METHODS: The EEG of 5 patients (ages 7-9) was recorded, spikes were submitted to blind decomposition in independent components (ICs) and those to source analysis (sLORETA), revealing the spike generators. Coherence analysis evaluated the dynamics of the components. RESULTS: Several ICs were recovered for posterior spikes in contrast to central spikes which originated a single one. Coherence analysis supports a model with epileptic activity originating near lateral occipital area and spreading to cortical temporal or parietal areas. CONCLUSIONS: Posterior spikes demonstrate rapid spread of epileptic activity to nearby lobes, starting in the lateral occipital area. In contrast, central spikes remain localized in the rolandic fissure. SIGNIFICANCE: Rapid spread of posterior epileptic activity in the Panayitopoulos type of occipital lobe epilepsy is responsible for the variable and poorly localized spike EEG. The lateral occipital cortex is the primary generator of the epileptic activity. PMID- 17398150 TI - Characteristic EEG differences between voluntary recumbent sleep onset in bed and involuntary sleep onset in a driving simulator. AB - OBJECTIVE: To investigate the differences in electroencephalographic (EEG) characteristics for voluntary sleep onset in bed sleeping and involuntary sleep onset in driving. METHODS: EEG measurement and analysis on 20 human subjects were conducted during recumbent bed sleeping as well as involuntary sleeping during a simulated driving platform. Each experiment was conducted on separate days. RESULTS: Vertex and spindle waves showed differing morphology under each condition. Vertex sharpness during recumbent sleep onset was significantly sharper than involuntary sleep onset during simulated driving. Sharpness of vertices from night-driving was significantly sharper than with day-driving. Triple conjoined vertex waves only occurred with voluntary recumbent sleep onset. A conjoined vertex spindle waveform was statistically associated with sleep onset whilst driving. CONCLUSIONS: This study has identified distinctive differences in EEG graphoelements during the sleep onset phase of recumbent and simulated driving conditions suggesting that EEG graphoelements are affected by cortical processes and vary according to the prevalent sleep condition. SIGNIFICANCE: This study could provide a further basis for developing safety alerting devices for the detection of sleep onset in the hope of improving driving safety. PMID- 17398151 TI - Motor evoked potential: a reliable and objective measure to document the functional consequences of multiple sclerosis? Relation to disability and MRI. AB - OBJECTIVE: In an attempt to analyze whether MEP can serve as a valid measure for evaluating neurological dysfunction in multiple sclerosis (MS), we related MEP to clinical and MRI measures. METHODS: Transcranial magnetic stimulation was applied in 52 MS patients to determine the central motor conduction time (CMCT) to the extremities. We calculated Z-scores for each CMCT (Zcmct) corrected for height. All patients underwent two clinical measurements and a MRI scan, of which T1 and T2 brain lesion volumes, brain volume, spinal cord volume and the number of T2 spinal cord lesions were derived. RESULTS: The clinical measurements correlated significantly with various Zcmct (Spearman correlation coefficients ranged from 0.29 to 0.53; p<0.05). The number of spinal cord lesions, brain T1 and T2 lesion volume and spinal cord volume correlated with various Zcmct (r=0.31-0.53; p<0.05). Linear regression analysis revealed that the clinical measurements were explained by Zcmct left leg and T1 lesion volume (adjusted R(2)=0.38). For one clinical measurement the number of spinal cord lesions was also included (adjusted R(2)=0.43). CONCLUSION: We found a relation between MEP, brain and spinal cord MRI measures, and two clinical measures. Moreover, a model for explaining disability in MS revealed that MEP measures provide information in addition to MRI measures. SIGNIFICANCE: This study suggests that MEP is a measure that might adequately reflect pathology and neurological dysfunction in MS. PMID- 17398152 TI - Effect of sleep on interictal spikes and distribution of sleep spindles on electrocorticography in children with focal epilepsy. AB - OBJECTIVE: To determine how sleep with central spindles alters the spatial distribution of interictal spike frequency in children with intractable focal seizures, and whether such children have spindles arising from the medial temporal region in addition to the frontal-central region. METHODS: Seventeen children (age: 7 months-17 years) were studied using extraoperative electrocorticography (ECoG). RESULTS: Overall spike frequency across the subdural electrodes was greater during sleep with central spindles compared to wakefulness. In 13 children showing at least 1 spike/min in an electrode, the spatial distribution of spike frequency was similar during wakefulness and sleep; in addition, the spike frequency was greater in the seizure onset zones compared to the non-onset areas, regardless of wakefulness or sleep. Spindles were identified in the medial temporal region during sleep with central spindles in all 17 children. CONCLUSION: Overall spike frequency may be increased by sleep with spindles, but the spatial distribution of spike frequency appears similar during wakefulness and sleep in children with intractable focal seizures. SIGNIFICANCE: Both awake and sleep ECoG may be useful to predict seizure onset zones in children with intractable focal epilepsy. Medial temporal spindles are present in some children with focal epilepsy. PMID- 17398153 TI - Event-based sonification of EEG rhythms in real time. AB - OBJECTIVE: To introduce a sound synthesis tool for human EEG rhythms that is applicable in real time. METHODS: We design an event-based sonification which suppresses irregular background and highlights normal and pathologic rhythmic activity. RESULTS: We generated sound examples with rhythms from well-known epileptic disorders and find stereotyped rhythmic auditory objects in single channel and stereo display from generalized spike-wave runs. For interictal activity, we were able to separate focal rhythms from background activity and thus enable the listener to perceive its frequency, duration, and intensity while monitoring. CONCLUSIONS: The proposed event-based sonification allows quick detection and identification of different types of rhythmic EEE events in real time and can thus be used to complement visual displays in monitoring and EEG feedback tasks. SIGNIFICANCE: The significance of the work lies in the fact that it can be implemented for on-line monitoring of clinical EEG and for EEG feedback applications where continuous screen watching can be substituted or improved by the auditory information stream. PMID- 17398154 TI - Cellular cardiomyoplasty in large myocardial infarction: can the beneficial effect be enhanced by ACE-inhibitor therapy? AB - BACKGROUND: Cellular cardiomyoplasty with bone marrow derived stromal (MSC) and mononuclear (BMNC) cells has been shown to improve performance of infarcted hearts. We performed a comparative study with MSC and BMNC and tested the hypothesis that captopril treatment could enhance the beneficial effect of cell therapy in large myocardial infarctions. METHODS: Male syngeneic Wistar rats underwent experimental infarction and were randomized to receive 1-3 x 10(6) MSC, 10(8) BMNC or vehicle (BSS group). Two additional groups were treated with captopril and received 1-3 x 10(6) MSC (Cap.MSC) or vehicle (Cap). RESULTS: The ejection fraction (EF%) of MSC and BMNC-treated rats was higher than in the BSS rats, eight weeks after transplantation (33.0+/-4.0, 34.0+/-2.0 and 20.0+/-2.0% respectively, P<0.01). Both captopril-treated groups improved EF% similarly. But only captopril plus MSC treatment almost restored cardiac function to control levels, 8 weeks after injection (60.50+/-5.40% vs. 41.00+/-4.50% in Cap.MSC and Cap respectively, P<0.05). Many DAPI-labelled cells were found in the scar tissue of the left ventricle only in the Cap.MSC group. CONCLUSIONS: Cell transplantation with both MSC and BMNC produced a similar stabilisation of heart function, but the success of the cell engraftment and the recovery of cardiac performance were dependent on concomitant treatment with captopril. PMID- 17398155 TI - Impact of heart failure and left ventricular function on long-term survival- report of a community-based cohort study in Taiwan. AB - BACKGROUND: There is little community-based information on heart failure (HF) prognosis in ethnic Chinese populations, in whom there is a low prevalence of coronary heart disease. AIMS: To study the impact of HF and left ventricular function on long-term all-cause mortality. METHODS AND RESULTS: This community based prospective cohort study included 2660 subjects (1215 men, 1445 women, mean age 54.4+/-11.9 years) over a 10 year follow-up period. The prevalence of HF was 5.5%. Hypertension was the most common factor related to HF. The five and ten year all-cause mortality was higher in the HF/preserved LVEF group (14.1% and 24.4%) and the HF/impaired LVEF group (29.2% and 48.2%) than in the HF-free group (6.0% and 14.6%, p<0.0001 for both). In multivariable Cox analyses, controlling for sex, LV mass, atrial fibrillation, hypertension, coronary heart disease, HF/preserved LVEF and HF/impaired LVEF were important predictors of all-cause mortality (p=0.007). CONCLUSIONS: Hypertension is a major heart failure related disease. HF and LV systolic dysfunction are associated with a significant increase in all-cause mortality in an ethnic Chinese population. PMID- 17398156 TI - Zentralblatt fur Bakteriologie - 100 years ago: the detection of the causative agent of syphilis, Treponema pallidum - the year after. PMID- 17398157 TI - Evaluation of pulsed-field gel electrophoresis and multi-locus sequence typing for the analysis of clonal relatedness among Bartonella henselae isolates. AB - Pulsed-field gel electrophoresis (PFGE) represents the gold standard among band based methods for the molecular typing of Bartonella henselae. SmaI and NotI have been frequently used for typing B. henselae by PFGE. However, their appropriateness for the analysis of genetic relatedness among B. henselae isolates has not been assessed systematically hitherto. Aim of the present study was to evaluate SmaI, NotI, and three additional endonucleases for typing B. henselae isolates by PFGE and to compare the PFGE results with multi-locus sequence typing (MLST) data. Twenty B. henselae isolates from different sources and geographic regions were analysed. PFGE analysis upon restriction with SmaI, ApaI, Eco52I, and XmaJI revealed six, five, four, and five different PFGE types, respectively, whereas restriction with NotI revealed 13 PFGE types. Five sequence types (STs) were obtained by MLST. The overall concordance between PFGE types obtained with SmaI, ApaI, Eco52I, XmaJI and STs was high. In contrast, NotI derived types did not correlate with other PFGE types or STs, indicating that NotI is not an appropriate enzyme for PFGE typing of B. henselae. By combining PFGE results obtained with SmaI, ApaI, Eco52I, XmaJI with STs, the isolates could be assigned to five distinct clonal lineages, including the clones Houston-1, Marseille, CAL-1, and Berlin-2. These data indicate that PFGE and MLST are discriminatory and reliable for molecular typing of B. henselae isolates to the strain level. Combination of PFGE and MLST may be useful for further epidemiological studies on B. henselae. PMID- 17398158 TI - Inhibitors of stress-activated protein/mitogen-activated protein kinase pathways. AB - The importance of stress-activated protein/mitogen-activated protein kinase (SAP/MAPK) pathway signalling (involving c-Jun-N-terminal kinase [JNK], extracellular signal-regulated kinase [ERK] and p38 kinase) in normal cellular proliferation, differentiation and programmed cell death has led to significant recent advances in our understanding of the role of SAP/MAPK signaling in inflammatory disorders such as arthritis and cardiovascular disease, cancer, and pulmonary and neurogenerative diseases. The discovery that several natural products such as resveratrol, tangeretin and ligustilide non-specifically inhibit SAP/MAPK signalling in vitro should now be logically extended to studies designed to determine how agents in these natural products regulate SAP/MAPK pathways in animal models of disease. A new generation of small-molecule SAP/MAPK inhibitors that demonstrate increasing specificity for each of the JNK, ERK and p38 kinase isoforms has shown promise in animal studies and could eventually prove effective for treating human diseases. Several of these compounds are already being tested in human subjects to assess their oral bioavailability, pharmacokinetics and toxicity. PMID- 17398159 TI - Epstein-Barr virus latent membrane protein 2A and autoimmunity. AB - Epstein-Barr virus (EBV) has been associated with autoimmune diseases for over 40 years. However, the mechanisms by which EBV might promote autoimmune development remain elusive. Many of the hypotheses for the means by which EBV might achieve this incorporate the idea that autoimmune responses are initially immune responses against EBV proteins that crossreact with endogenous human proteins. However, recent evidence using transgenic mouse models suggests that B cells expressing the EBV-encoded protein latent membrane protein 2A (LMP2A) bypasses normal tolerance checkpoints and enhances the development of autoimmune diseases. Evidence from transgenic mouse models supports a paradigm in which LMP2A could promote autoimmune development. This novel model provides a framework to test potential mechanisms by which EBV could promote the development of autoimmune responses and might enable the identification of strategies to treat EBV associated autoimmune diseases. PMID- 17398160 TI - Primer on risk management for the gynaecological laparoscopist. AB - The gynaecologist practising operative laparoscopy should be seen as part of a team that actively promotes patient safety, minimizing risks and optimizing outcomes. Building a culture of safety which focuses on proactive initiatives to manage risk and remove individual 'blame' should be an integral part of any operative laparoscopy unit. Thus, when adverse clinical incidents or outcomes occur, reporting of such events is encouraged and seen to be acceptable behaviour within the framework of complete patient care. By recognizing and analysing adverse outcomes, the team can develop strategies to prevent or manage a recurrence of such events. Implementing systems or solutions to prevent harm to patients is the cornerstone of any risk management programme. In this review, we discuss the development and implementation of risk management strategies in the clinical setting, and in particular how this applies to operative laparoscopy. PMID- 17398161 TI - Patient safety in women's health care: a framework for progress. AB - Patient safety research is hampered by lack of a clear taxonomy and difficulty in detecting errors. Preventable adverse events occur in medicine because of human fallibility, complexity, system deficiencies and vulnerabilities in defensive barriers. To make medicine safer there needs to be a culture change, beginning with the leadership. Latent systems deficiencies must be identified and corrected before they cause harm. Defensive barriers can be improved to intercept errors before patients are harmed. Strategies include: (1) providing leadership at all levels; (2) respecting human limits in equipment and process design; (3) functioning collaboratively in a team model with mutual respect; (4) creating a learning environment where errors can be analyzed without fear of retribution; and (5) anticipating the unexpected with analysis of high-risk processes and well designed contingency plans. The ideal of a 100% safe health-care system is unattainable, but there must be continual improvement. PMID- 17398162 TI - Variability in expression of Bothrops insularis snake venom proteases: an ontogenetic approach. AB - Bothrops insularis is a threatened snake endemic to Queimada Grande Island, southern coast of Sao Paulo, Brazil, and the occurrence of sexual abnormalities in males, females and intersexes (females with functional ovaries and rudimentary hemipenis) has been reported in this population. The aim of this study was to identify ontogenetic shifts in protease expression of offspring of captive-bred B. insularis. Three neonates from a single litter were maintained at the facilities of Laboratory of Herpetology, Institute Butantan, for 41 months. The snakes were individually milked and venoms were analyzed both by SDS-PAGE, under reducing conditions, and for biochemical activities. The venoms from the mother and from a pool of adult specimens were used as references. In regard to the electrophoretic patterns, common bands were identified mainly between 14 and 50 kDa among snakes. The occurrence of proteolytic activity was noticed predominantly between 27 and 45 kDa in zymograms. Inhibitory assays with 1,10 phenantroline (10 mM) and PMSF (5 mM) showed that venoms possessed both metalloproteases and serine proteases. Venoms of young specimens showed a higher coagulant activity than those of adults, especially upon factors X and II. All venoms presented fibrino(geno)lytic activity, degrading Aalpha and Bbeta chains of fibrinogen, and lysing fibrin plate. These findings can reflect important individual, ontogenetic and sexual differences on venom composition and are likely correlated with diet habits of this species. PMID- 17398163 TI - Creating a culture of research. AB - There is widespread concern about the ability of the current model of clinical research to keep pace with the growing need for testing new approaches to disease management and prevention. In response, important innovations are emerging in critical areas of research infrastructure and practice. However, success in fulfilling the promise of clinical research will also require a fundamental shift in the relationship between the clinical-research enterprise and all segments of society. In this article, we outline proposals to help create the necessary recognition and engage participation by patients, clinicians, health-care delivery systems, and the research community to establish the long-lasting growth needed for achieving the full potential of clinical research. PMID- 17398164 TI - Comparative transcript profiles of cell cycle-related genes in mouse primordial germ cells, embryonic stem cells and embryonic germ cells. AB - We used cDNA array to compare the relative transcript levels of 96 cell cycle related genes in mouse primordial germ cells (PGCs), embryonic germ (EG) cells and embryonic stem (ES) cells. Among 38 genes of the G1 phase analysed, Ccnd3 (CyclinD3), Cdkn1c (p57(kip2)), Rb1, and Tceb1l (Skip1-like) were expressed at significantly higher levels in PGCs than in EG and ES cells; Ccnd1 (CyclinD1) was more abundant in EG cells than in PGCs. Except for higher mRNA levels of Ccng (CyclinG1) in EG and ES cells in comparison to PGCs, no difference among 20 genes of the S and 12 genes of G2/M phases was found. Less than half of the 26 genes regarded as DNA damage checkpoint/Trp53/Atm pathway genes showed significant transcript levels in all three cell populations. Among these, the transcript levels of Ube1x and Atm were significantly higher in PGCs than in EG and ES cells while that of Ube3a was higher in these latter. In addition, relatively high mRNA levels of Timp3 characterizes EG cells while transcripts of this gene were very low in PGCs and barely detectable in ES cells. With the exception of Tceb1l, differential transcript levels found in the cDNA array assay were confirmed by real time RT-PCR. Using this method, we also analysed the transcripts of two genes not present in the cDNA array: c-myc, known to be critical for the control of cell cycle in many cell types, and Eras, specifically expressed in ES cells and involved in the control of ES cell proliferation and their tumorigenic properties. While c-myc transcripts were present at similar levels in all three cell types examined, Eras was expressed at high levels in ES cells (10-fold) and even more so in EG cells (almost 40-fold) in comparison to PGCs. Taken together, these results indicate that despite similarities between PGCs and ES or EG cells, their cell cycles are differently regulated. In particular, it appears that PGCs, like most mitotic cells, possess a more regulatable control of G1 phase than EG and ES cells. Moreover, our data provide useful clues for further studies aimed at identifying cell cycle genes critical for PGC growth and their transformation in tumorigenic cells. PMID- 17398165 TI - Improved differentiation of Mycobacterium tuberculosis strains, including many Beijing genotype strains, using a new combination of variable number of tandem repeats loci. AB - Variable number of tandem repeats (VNTR) typing was done on 230 Mycobacterium tuberculosis strains, including 41 strains isolated from 17 groups of epidemiologically linked patients. By PCR amplification, 185 (80.4%) of the 230 strains were Beijing genotype strains. VNTR typing was performed using the 15 loci proposed as a standard set by Supply et al. [Supply, P., Allix, C., Lesjean, S., Cardoso-Oelemann, M., Rusch-Gerdes, S., Willery, E., Savine, E., de Haas, P., van Deutekom, H., Roring, S., Bifani, P., Kurepina, N., Kreiswirth, B., Sola, C., Rastogi, N., Vatin, V., Gutierrez, M.C., Fauville, M., Niemann, S., Skuce, R., Kremer, K., Locht, C., van Soolingen, D., 2006. Proposal for standardization of optimized mycobacterial interspersed repetitive unit-variable-number tandem repeat typing of Mycobacterium tuberculosis. J. Clin. Microbiol. 44, 4498-4510], and cluster analyses of these data were done. By the VNTR typing with the proposed 15 loci, strains having low similarity values by restriction fragment length polymorphism (RFLP) analysis were clustered. Use of a supplemental9 loci, proposed as a high-resolution tool, with the 15 loci showed that strains with low similarity by RFLP analysis were still clustered. Twelve VNTR loci were selected based on previously reported discriminatory index (DI) values and used with the proposed 15 loci for better differentiation by VNTR typing. When eight loci with higher DI values were used with the 15 loci, there were no clusters, including strains with low RFLP similarity. The15 loci and eight additional loci decreased the numbers of clustered strains isolated from epidemiologically unlinked patients significantly compared to using only the 15 loci. Among all tested loci, obvious differences of DI values were observed for 8 loci (miru10, miru16, miru39, Mtub29, Mtub30, QUB11a, QUB26, and QUB1895) of RD105 lineage strains compared to those of other lineage strains. These results suggest that the proposed VNTR typing method cannot be used as a routine epidemiological tool in areas where Beijing genotype strains are prevalent. Several VNTR loci should be added to the proposed method based on differences in polymorphism of VNTR loci among Beijing genotype lineages. PMID- 17398166 TI - Direct electrochemistry and electrocatalysis of hemoglobin entrapped in semi interpenetrating polymer network hydrogel based on polyacrylamide and chitosan. AB - Semi-interpenetrating polymer network (semi-IPN) hydrogel based on polyacrylamide (PAM) and chitosan was prepared to immobilize redox protein hemoglobin (Hb). The Hb-PAM-chitosan hydrogel film obtained has been investigated by scanning electron microscopy (SEM) and UV-VIS spectroscopy. UV-VIS spectroscopy showed that Hb kept its secondary structure similar to its native state in the Hb-PAM-chitosan hydrogel film. Cyclic voltammogram of Hb-PAM-chitosan film-modified glass carbon (GC) electrode showed a pair of well-defined and quasi-reversible redox peaks for Hb Fe(III)/Fe(II), indicating that direct electron transfer between Hb and GC electrode occurred. The electron-transfer rate constant was about 5.51 s(-1) in pH 7.0 buffers, and the formal potential (E degrees ') was -0.324 V (vs. SCE). The dependence of E degrees ' on solution pH indicated that one-proton transfer was coupled to each electron transfer in the direct electron-transfer reaction. Additionally, Hb in the semi-IPN hydrogel film retained its bioactivity and showed excellent electrocatalytic activity toward H(2)O(2). The electrocatalytic current values were linear with increasing concentration of H(2)O(2) in a wide range of 5-420 microM. The unique semi-IPN hydrogel would have wide potential applications in direct electrochemistry, biosensors and biocatalysis. PMID- 17398167 TI - Immobilization of E. coli bacteria in three-dimensional matrices for ISFET biosensor design. AB - In recent years, cell-based biosensors (CBBs) have been very useful in biomedicine, food industry, environmental monitoring and pharmaceutical screening. They constitute an economical substitute for enzymatic biosensors, but cell immobilization remains a limitation in this technology. To investigate into the potential applications of cell-based biosensors, we describe an electrochemical system based on a microbial biosensor using an Escherichia coli K 12 derivative as a primary transducer to detect biologically active agents. pH variations were recorded by an ion-sensitive field effect transistor (ISFET) sensor on bacteria immobilized in agarose gels. The ISFET device was directly introduced in 100 ml of this mixture or in a miniaturized system using a dialysis membrane that contains 1 ml of the same mixture. The bacterial activity could be detected for several days. The extracellular acidification rate (ECAR) was analyzed with or without the addition of a culture medium or an antibiotic solution. At first, the microorganisms acidified their micro-environment and then they alkalinized it. These two phases were attributed to an apparent substrate preference of bacteria. Cell treatment with an inhibitor or an activator of their metabolism was then monitored and streptomycin effect was tested. PMID- 17398168 TI - Transcription-associated mutagenesis in yeast is directly proportional to the level of gene expression and influenced by the direction of DNA replication. AB - A high level of transcription has been associated with elevated spontaneous mutation and recombination rates in eukaryotic organisms. To determine whether the transcription level is directly correlated with the degree of genomic instability, we have developed a tetracycline-regulated LYS2 reporter system to modulate the transcription level over a broad range in Saccharomyces cerevisiae. We find that spontaneous mutation rate is directly proportional to the transcription level, suggesting that movement of RNA polymerase through the target initiates a mutagenic process(es). Using this system, we also investigated two hypotheses that have been proposed to explain transcription-associated mutagenesis (TAM): (1) transcription impairs replication fork progression in a directional manner and (2) DNA lesions accumulate under high-transcription conditions. The effect of replication fork progression was probed by comparing the mutational rates and spectra in yeast strains with the reporter gene placed in two different orientations near a well-characterized replication origin. The effect of endogenous DNA damage accumulation was investigated by studying TAM in strains defective in nucleotide excision repair or in lesion bypass by the translesion polymerase Polzeta. Our results suggest that both replication orientation and endogenous lesion accumulation play significant roles in TAM, particularly in terms of mutation spectra. PMID- 17398169 TI - Novel CFTR gene mutation in a patient with CBAVD. AB - We report a novel mutation detected in a 33 year old Chinese man with congenital bilateral absence of the vas deferens (CBAVD), a past history of pulmonary meliodosis infection and a past history of bronchiolitis obliterans organising pneumonia. A novel splice site mutation in intron 6b (1001+5 G-->A) in the homozygous state was identified, and was predicted to lead to inefficient splicing. He was also homozygous at all intragenic and flanking polymorphic markers. Quantitative realtime PCR analysis showed that there were 2 copies of the CFTR gene present, ruling out the possibility of a deletion, and strongly suggesting the possibility of uniparental isodisomy involving at least a part of chromosome 7. PMID- 17398170 TI - Mutational analysis of O-acetylserine (thiol) lyase conducted in yeast two-hybrid system. AB - Cysteine biosynthesis, achieved by the sequential reaction of two enzymes, serine acetyltransferase and O-acetylserine (thiol) lyase (OASTL), represents the final step of sulfur assimilation pathway in plants and bacteria. The two enzymes form a bi-enzymatic cysteine synthase complex through specific protein-protein interactions. To identify the amino acids important for cysteine synthase complex formation, several mutations in bacterial OASTL were designed. Effects of mutagenesis were verified in a yeast two-hybrid model that allowed monitoring both, protein-protein interactions and the enzymatic activity of OASTL. PMID- 17398171 TI - Cellular response to oxygen containing biomedical polymers modified by Ar and He implantation. AB - Ion beam modification is an attractive way to adapt the response of a biopolymer surface with the view to modifying cellular processes. In this work we performed Ar and He implantations into three oxygen-containing biomedical polymers: polycaprolactone (PCL), poly(ethylene glycol) (PEG) and poly(methyl methacrylate) (PMMA). An ion energy of 25keV was selected on the basis of singularities observed in simulated implantations. The implantations were carried out with fluences of 5x10(13) cm(-2) considering also the ion current density as a source of differentiated damage. The modification of the polymer structure and composition was assayed by Fourier transform infrared spectroscopy, which confirmed the selectivity of the ion current density in producing polymer film damage. Biomedical assays denoted lack of structural stability on the PMMA surfaces. Surface analysis of proteins adsorbed from fetal bovine serum on ion beam-modified PEG were realized by quartz-crystal microbalance with dissipation, which supported the film stabilization and anti-fouling behaviour of the films. On the other hand, protein adsorption studies on micropatterned PCL surfaces were performed by time-of-flight secondary ion mass spectroscopy and revealed a clear enhancement of protein immobilization in ion-beam-modified areas. The response of human mesenchymal stem cells to the surfaces was observed to depend on the biopolymer characteristics, showing adhesion inhibition onto He-modified PEG and specially enhanced colonization onto He-irradiated PCL. PMID- 17398173 TI - Targeted bone remodeling involves BMU steering as well as activation. AB - Microdamage removal is an important function of bone remodeling. Experiments have repeatedly shown that remodeling of cortical bone by Basic Multicellular Units (BMUs) is initiated in response to microdamage, and this has become known as "targeted remodeling". This paper considers the possibility that microdamage is not only able to activate new BMUs, but may also attract or "steer" existing BMUs as they continue to tunnel through the bone matrix. An initial analysis of the relationship of between mean microcrack length and BMU resorption space density in cortical bone indicates that BMUs have an effective area about 40 times greater than their actual cross-section. Interpreting this as evidence that the osteoclasts in a tunneling BMU are able to sense and steer toward microdamage, a model is developed for "BMU steering" based on the hypothesis that osteoclasts are guided not only in the principal stress direction, as proposed by Burger et al. (Burger, E.H., Klein-Nulend, J., Smit, T.H. Strain-derived canalicular fluid flow regulates osteoclast activity in a remodelling osteon-a proposal. J. Biomech 36 (2003) 1453-1459), but also toward microdamage, depending on its proximity. PMID- 17398174 TI - Changes in mRNA gene expression during growth in the femoral head of the young rat. AB - The rate of physeal growth slows as an animal matures with changes in mRNA gene expression due to the altered cellular activity. To measure the change in gene expression during the juvenile growth period, the femoral head, enclosing the proximal femoral physis, primary spongiosa, and articular cartilage, was collected from both femora of 16 female Sprague-Dawley rats between 4 and 10 weeks of age. One femur of each rat had had a mid-diaphyseal femoral fracture at 4 weeks of age. RNA was extracted and hybridized to 16 Affymetrix Rat Genomic 230 2.0 GeneChip microarrays with probe sets for 31,000 genes of which 18,200 were expressed. Of these, 8002 genes had a significant change in gene expression during growth, about half increasing and half decreasing. These changes included up-regulation with time of genes related to cartilage, blood vessels, osteoprotegerin, osteomodulin, and most ribosomal proteins. There was down regulation with maturity of genes related to bone, growth-promoting cytokines, G proteins, GTPase-mediated signal transduction factors, cytokine receptors, mitosis, integrin-linked kinase, and the cytoskeleton. In summary, the slowing of growth with maturity was associated with changes in mRNA gene expression in the femoral head for a large number of genes. These changes in gene expression between young and mature rats suggest factors which are important for the support of the rapid linear growth during early life. PMID- 17398175 TI - Mechanical force-induced midpalatal suture remodeling in mice. AB - Mechanical stress is an important epigenetic factor for regulating skeletal remodeling, and application of force can lead to remodeling of both bone and cartilage. Chondrocytes, osteoblasts and osteoclasts all participate and interact with each other in this remodeling process. To study cellular responses to mechanical stimuli in a system that can be genetically manipulated, we used mouse midpalatal suture expansion in vivo. Six-week-old male C57BL/6 mice were subjected to palatal suture expansion by opening loops with an initial force of 0.56 N for the periods of 1, 3, 5, 7, 14 or 28 days. Periosteal cells in expanding sutures showed increased proliferation, with Ki67-positive cells representing 1.8+/-0.1% to 4.5+/-0.4% of total suture cells in control groups and 12.0+/-2.6% to 19.9+/-1.2% in experimental/expansion groups (p<0.05). Starting at day 1, cells expressing alkaline phosphatase and type I collagen were seen. New cartilage and bone formation was observed at the oral edges of the palatal bones at day 7; at the nasal edges only bone formation without cartilage appeared to occur. An increase in osteoclast numbers suggested increased bone remodeling, ranging from 60 to 160% throughout the experimental period. Decreased Saffranin O staining after day 3 suggested decreased proteoglycan content in the secondary cartilage. Micro-CT showed a significant increase in maxillary width at days 14 and 28 (from 2334+/-4 microm to 2485+/-3 microm at day 14 and from 2383+/-5 microm to 2574+/-7 microm at day 28, p<0.001). The suture width was increased at days 14 and 28, except in the oral third region at day 28 (from 48+/-5 microm to 36+/-4 microm, p<0.05). Bone volume/total volume was significantly reduced at days 14 and 28 (50.2+/-0.7% vs. 68.0+/-3.7% and 56.5+/-1.0% vs. 60.9+/-1.3%, respectively, p<0.05), indicative of increased bone marrow space. These findings demonstrate that expansion forces across the midpalatal suture promote bone resorption through activation of osteoclasts and bone and cartilage formation via increased proliferation and differentiation of periosteal cells. Mouse midpalatal suture expansion would be useful in further studies of the ability of mineralized tissues to respond to mechanical stimulation. PMID- 17398176 TI - Inflammatory markers, angiographic severity of coronary artery disease, and patient outcome. AB - Serum levels of high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL 6), and tumor necrosis factor-alpha (TNF-alpha) have been shown to be predictors of adverse outcomes in patients with coronary artery disease (CAD). We hypothesized that measurement of inflammatory markers could predict atherosclerotic burden and major adverse cardiac events (MACEs). We prospectively measured hs-CRP, IL-6, and TNF-alpha in 249 patients who were admitted with acute chest pain and underwent coronary angiography. We analyzed the relation between serum levels of inflammatory markers and angiographic severity of CAD. A follow up at 6 months was conducted to assess MACEs, defined as a cumulative of myocardial infarction, all-cause death, or coronary revascularization (percutaneous coronary intervention or coronary artery bypass surgery). After adjusting for conventional CAD risk factors (age, gender, diabetes, hypertension, smoking, and hypercholesterolemia), there was no association between inflammatory markers (hs-CRP, IL-6, and TNF-alpha) and angiographic severity of CAD. There was a significant positive correlation between age, male gender, diabetes mellitus, and hypercholesterolemia with atherosclerotic burden determined by angiography. There was no significant positive association between MACEs and hs-CRP, IL-6, or TNF-alpha level in unadjusted and adjusted models. In conclusion, in patients hospitalized with chest pain, we found no association of serum levels of hs-CRP, IL-6, or TNF-alpha with coronary atherosclerotic burden or MACEs at 6 months after adjustment for traditional CAD risk factors. PMID- 17398177 TI - Clinical impact of metabolic syndrome and its additive effect with smoking on subsequent cardiac events after acute myocardial infarction. AB - Little information is available regarding the clinical effect of metabolic syndrome (MS) or its combined effect with smoking on subsequent cardiac events after acute myocardial infarction (AMI). To examine whether MS independently predicts cardiac events (cardiac death and nonfatal reinfarction) and to assess the combined effect of MS and smoking on cardiac events after AMI, we studied 3,858 survivors of AMI registered in the Osaka Acute Coronary Insufficiency Study (OACIS). During a median follow-up of 725 days, the incidence of cardiac events was higher in patients with MS than in those without MS (p=0.021). After adjustment for baseline characteristics, MS was an independent predictor of cardiac events after AMI (hazard ratio [HR] 1.480, 95% confidence interval [CI] 1.128 to 1.942, p=0.005). Compared with patients who did not have MS and were nonsmokers, the adjusted HR for cardiac events was 2.868 (95% CI 1.573 to 5.227, p=0.001) in patients with MS who continued smoking after AMI. Cessation of smoking after AMI was associated with a significantly lower risk of cardiac events in patients with MS (HR 0.485, 95% CI 0.281 to 0.837, p=0.009) but not in patients without MS (HR 0.618, 95% CI 0.330 to 1.157, p=0.132). In conclusion, MS is independently associated with an increased risk of cardiac events after AMI. Smoking has an additive adverse effect on cardiac events after AMI, and cessation of smoking is beneficial and strongly recommended for AMI, especially in patients with MS. PMID- 17398178 TI - Serum complement C3/C4 ratio, a novel marker for recurrent cardiovascular events. AB - Acute coronary syndrome is an inflammatory disease, during which the complement cascade is activated. We assessed the complement C3 and C4 concentration ratio (C3/C4 ratio) in serum as a potential measurement to predict cardiovascular attacks. Patients with acute coronary syndrome (n=148) were followed after an initial attack for subsequent ischemic cardiovascular events (composite end point of death, myocardial infarction, recurrent unstable angina, or stroke). During the follow-up period (average 555 days), 44 patients met an end point. Blood samples were taken at hospitalization, 1 week, 3 months, and 1 year after hospital admission. Serum complement C3 and C4 concentrations and the C3/C4 ratio were analyzed. Patients with an end point had, throughout the follow-up period, a higher C3/C4 ratio than patients without these end points (repeated measures analysis of variance, p=0.007). When all traditional cardiovascular risk factors and other potential confounding factors were included in a Cox multivariate logistic regression survival analysis, the C3/C4 ratio emerged as the novel risk factor for any new cardiovascular event (odds ratio 1.33, 95% confidence interval 1.08 to 1.63, p=0.007). When the C3/C4 ratio was divided into 4 quartiles, 24% in quartiles 1 and 2 (lowest) and 48% in quartile 4 (highest) had end points during follow-up (odds ratio 3.04, 95% confidence interval 1.27 to 7.29, p=0.01). In conclusion, increased serum C3/C4 ratio is a readily available and novel marker for recurrent cardiovascular events in acute coronary syndrome. The relative increase in serum C3 protein and decrease in C4 protein could explain changes in the C3/C4 ratio. PMID- 17398179 TI - Comparison between angiography and fractional flow reserve versus single-photon emission computed tomographic myocardial perfusion imaging for determining lesion significance in patients with multivessel coronary disease. AB - We hypothesized that myocardial perfusion imaging (MPI) would fail to identify all vascular zones with the potential for myocardial ischemia in patients with multivessel coronary disease (MVD). MPI is based on the concept of relative flow reserve. The ability of these techniques to determine the significance of a particular stenosis in the setting of MVD is questionable. Fractional flow reserve (FFR) can determine the significance of individual stenoses. Thirty-six patients with disease involving 88 arteries underwent angiography, FFR, and MPI. FFR was performed using a pressure wire with hyperemia from intracoronary adenosine. Myocardial perfusion images were analyzed quantitatively and segments assigned to a specific coronary artery. The relation between FFR and perfusion was determined for each vascular zone. Of the 88 vessels, the artery was occluded (n=20) or had an abnormal FFR70 years, final Thrombolysis In Myocardial Infarction grade<3 flow, history of peripheral vascular disease, and ejection fraction to be the strongest predictors of mortality at 12 months. In conclusion, our data show that obese patients with AMI have a lower risk for in-hospital, 6 month, and 12-month mortality and cardiovascular events than patients with a normal BMI. PMID- 17398182 TI - Safety and efficacy of the FilterWire EZ in acute ST-segment elevation myocardial infarction. AB - Primary percutaneous coronary intervention (PCI) for ST-segment elevation myocardial infarction (STEMI) achieves a high epicardial reperfusion rate; however, it is often suboptimal in achieving myocardial reperfusion due to distal embolization of atherothrombotic particles. The present study assessed whether the capture of embolic particles during PCI would improve myocardial reperfusion outcome. In a multicenter, prospective, randomized, controlled study, 100 patients with STEMI and coronary angiographic evidence of thrombotic occlusion were randomly assigned to PCI using the FilterWire EZ (n=51) or a control group (n=49) using regular guidewires. The FilterWire EZ was successfully delivered across the lesion in 84% of patients in the FilterWire EZ group. Primary efficacy end points, including markers of epicardial (Thrombolysis In Myocardial Infarction grade flow) and myocardial reperfusion (myocardial blush score and percent early resolution of ST-segment elevation), did not differ between the 2 study groups. Further, 60- and 90-minute percent ST-segment resolutions were identical in the 2 groups. In a subgroup analysis, a blush score of 3 was achieved in 94% of patients in whom the filter's landing zone was in a vessel diameter>2.5 mm compared with only 55% in those with smaller vessel diameter (p=0.04). This corresponds to a better debris capture in filters located in large versus small vessels (p=0.08). In conclusion, in patients with STEMI, use of the FilterWire EZ as an adjunct to primary PCI did not improve angiographic or electrocardiographic measurements of reperfusion compared with conventional PCI only. PMID- 17398183 TI - Frequency of slow coronary flow following successful stent implantation and effect of Nitroprusside. AB - Nitroprusside (NTP) is used for the treatment of slow coronary flow (SCF) after coronary interventions. The wide variation in dosage, route, and timing of its administration in the reported studies prevents an objective assessment of its efficacy. We report the incidence and response to a standardized NTP protocol of SCF after successful stent implantation. Selective intracoronary administration of incremental doses (initial bolus of 80 microg incremented by 40 microg) of NPT was assessed in 21 patients who developed SCF in a series of 2,212 consecutive patients who underwent successful stent placement from January to October 2005. SCF was observed only in patients treated for acute myocardial infarction (AMI; 11.5%, 12 of 105) or saphenous vein graft (SVG) stenosis (8.2%, 9 of 109). An intracoronary bolus of nitroglycerin did not restore normal Thrombolysis In Myocardial Infarction (TIMI) flow in any patient. The first 80-microg dose of NTP restored normal TIMI flow in 58% of patients (7 of 12) with AMI and in 44% of patients (4 of 9)with SVG stenosis. The maximal dose (120/160 microg) restored normal TIMI flow in all remaining patients with AMI but in only 1 additional patient with SVG stenosis. At the end of the procedure, the percent decrease in corrected TIMI frame count was significantly larger in patients with AMI (-44+/ 10%) than in those with SVG stenosis (-24+/-16%, p=0.02). In a large consecutive series of successful stent procedures, SCF was found only in patients with ST elevation AMI (11.5%) or with a stenosed SVG (8.2%). In conclusion, the standardized protocol of intracoronary NTP administration succeeded in normalizing SCF in all patients with AMI but in only 5 of 9 patients with SVG stenosis. This latter subgroup requires other therapeutic strategies. PMID- 17398184 TI - Different thresholds of myocardial ischemia in ramp and standard bruce protocol exercise tests in patients with positive exercise stress tests and angiographically demonstrated coronary arterial narrowing. AB - Gradual instead of abrupt increases in workload favor a more physiologic response in terms of hemodynamic and gas exchange parameters. Therefore, we investigated whether myocardial ischemia is attenuated with a ramp compared with a standard Bruce exercise protocol in patients with coronary artery disease. We compared electrocardiographic ischemic parameters on the standard Bruce protocol treadmill and the individualized ergocycle ramp protocol in 18 men with coronary artery disease and a reproducible ischemic electrocardiographic exercise test. Oxygen consumption (VO2), ischemic threshold (rate-pressure product [RPP]=systolic blood pressure x heart rate at 1-mm ST-segment depression), and maximum ST-segment depression corresponding to the highest RPP common to the 2 tests were determined. Ischemic threshold was higher with the ramp than with the Bruce protocol (23,420+/-5,732 vs 20,018+/-3,542 beats.min/mm Hg, p=0.007). Peak RPP was higher during the ramp than during the Bruce protocol (28,492+/-6,450 vs 25,519+/-6,067 beats.min/mm Hg, respectively, p=0.02) despite similar peak VO2 (25.59+/-5.05 vs 26.39+/-4.65 mlO2.kg-1.min-1, respectively, p=0.6). Maximum ST segment depression corresponding to the highest RPP common to the 2 tests was less with the ramp than with the Bruce protocol (-1.2+/-0.9 vs -1.9+/-0.7 mm, p=0.003). In conclusion, exercise-induced myocardial ischemia is markedly attenuated on the more gradually increasing workload of the individualized ramp ergocycle compared with the standard Bruce treadmill protocol. This effect is unexplained by energy expenditure (VO2) or myocardial work (RPP) and is consistent with a "warm-up" ischemic mechanism. PMID- 17398185 TI - Usefulness of 64-slice multidetector computed tomography in diagnostic triage of patients with chest pain and negative or nondiagnostic exercise treadmill test result. AB - The usefulness of 64-slice multidetector coronary computed tomography (MDCT) in a diagnostic triage of 100 consecutive patients (age 55.8+/-11.6 years; 57% men) with chest pain suspected to be ischemic in origin and a negative or nondiagnostic exercise treadmill test (ETT) result was examined. None of the patients had previously known coronary artery disease (CAD). MDCT showed obstructive (>or=50%) CAD in 29 patients; 13 of 59 patients (22%) with a negative and 16 of 41 patients (39%) with a nondiagnostic ETT result. High-risk (left main and/or 3-vessel) CAD was present in 3.3% of patients with a negative and 4.9% with a nondiagnostic ETT result. The 29 patients with obstructive CAD on MDCT had a higher mean Agatston calcium score (221+/-402 vs 40+/-77 U, p<0.001). Invasive coronary angiography confirmed MDCT findings in 26 of 29 patients (positive predictive value 90%) and 45 of 54 stenotic segments (83%) in a per-segment analysis. For the 71 patients without obstructive CAD on MDCT, clinically driven invasive angiography detected CAD in 1 of 15 patients (1 false-negative MDCT result) and 2 of another 5 patients who were referred for invasive angiography later during a 12-month follow-up period. In the remaining 51 patients, MDCT findings effectively allowed exclusion of obstructive CAD, and there were no major adverse clinical events during follow-up. In conclusion, in patients with chest pain possibly ischemic in origin, no previously known CAD, and a negative or nondiagnostic ETT result, contrast-enhanced 64-slice MDCT scanning was a useful tool to provide direct noninvasive coronary angiography and rapidly advance diagnostic triage. PMID- 17398186 TI - Self-reported use of complementary and alternative medicine in patients with previous acute coronary syndrome. AB - Complementary and alternative medicine (CAM) use is common in patients with cardiovascular disease. Although numerous efforts have sought to understand CAM types and the prevalence of CAM, whether patients preferentially use CAM instead of evidence-based therapies is unknown. Self-reported use of CAM and evidence based therapies in a prospective registry of hospitalized patients with acute coronary syndrome from March 1, 2001 to October 31, 2002 were examined. Poisson regression models were used to assess whether CAM use was independently associated with lower rates of aspirin, beta-blocker, and statin use in 596 patients with established coronary artery disease (CAD). Overall, CAM use was 19% in patients with CAD. Higher proportions of patients who used CAM were non Caucasian (31% vs 12%), uninsured (12% vs 7%), economically burdened (58% vs 29%), and had depression (13% vs 6%, p<0.05 for all). Patients who used CAM were more likely to use beta blockers (64% vs 46%, p=0.008) and as likely to use aspirin (73% vs 74%, p=0.90) and statins (71% vs 68%, p=0.76) as non-CAM users. Adjusting for demographic and clinical factors did not change results (CAM users: RR 1.27, 95% confidence interval [CI] 1.01 to 1.60 for using beta blockers, RR 0.97, 95% CI 0.85 to 1.11 for using aspirin, and RR 1.05, 95% CI 0.87 to 1.28 for using statins). In conclusion, although CAM users with established CAD have worse socioeconomic status than nonusers, we found no evidence that they were less compliant with evidence-based therapies. PMID- 17398187 TI - Usefulness of ST depression with T-wave inversion in leads V(4) to V(6) for predicting one-year mortality in non-ST-elevation acute coronary syndrome (from the Electrocardiographic Analysis of the Global Use of Strategies to Open Occluded Coronary Arteries IIB Trial). AB - ST-segment depression (ST-D) on the admission electrocardiogram of patients with non-ST-elevation acute coronary syndromes (NSTEACSs) is associated with higher mortality. However, few studies have evaluated the effect of location of ST-D and T-wave polarity on long-term prognosis of patients with NSTEACS. Electrocardiographic (ECG) and clinical data from 6,770 patients with NSTEACS randomly assigned in the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO) IIB trial were analyzed retrospectively. One-year mortality was correlated with location of ST-D (leads I and aVL; II, III, and aVF; V1 to V3; or V4 to V6) and T-wave polarity. ST-D in any of the ECG locations was associated with higher mortality compared with patients without ST-D. Patients with ST-D and T-wave inversion in leads V4 to V6 had the highest 1-year mortality rate of all groups (16.2%), significantly higher compared with patients with ST-D without T wave inversion in those leads (9.0%, p=0.001). Logistic regression analysis showed that age, hyperlipidemia, Killip class>I, history of myocardial infarction, history of heart failure, history of angina pectoris, systolic blood pressure, heart rate, sum of ST-D (odds ratio 1.061, 95% confidence interval 1.035 to 1.087, p<0.001), and ST-D with T-wave inversion in leads V4 to V6 (odds ratio 1.374, 95% CI 1.023 to 1.844, p=0.035) were independent predictors of 1 year mortality. Conversely, ST-D without T-wave inversion in leads V4 to V6 or other ECG presentations were not independent predictors of high 1-year mortality. In conclusion, ST-D with T-wave inversion in leads V4 to V6 on the admission electrocardiogram in patients with NSTEACS identifies those with higher 1-year mortality than for patients with any other ECG presentation. PMID- 17398188 TI - Prognostic value of transient and sustained increase in in-hospital creatinine on outcomes of patients admitted with acute coronary syndrome. AB - A history of renal insufficiency or increased creatinine level on admission is associated with poor outcomes in patients with acute coronary syndrome (ACS). This study sought to determine whether in-hospital worsening of renal function, either transient or sustained, is an independent risk factor for 6-month mortality in patients admitted with ACS. A total of 1,417 patients admitted with ACS from June 2000 to May 2003 were reviewed. Patients were classified into 3 groups. Group I included patients with an increase in creatinine during hospitalization of 0.5 mg/dl that resolved by discharge. Group III included patients with an increase in creatinine of >0.5 mg/dl that did not resolve. The primary end point was 6-month mortality from any cause. Patients in groups II and III had higher 6-month mortality rates (27% and 23%, respectively; both p<0.001) compared with patients in group I (7.4%). After adjustment for known risk factors, a transient increase in creatinine remained a significant independent predictor of 6-month mortality (odds ratio 2.07, 95% confidence interval 1.14 to 3.76), although a sustained increase in creatinine showed a trend (odds ratio 1.58, 95% confidence interval 0.68 to 3.70). In conclusion, independent of a history of renal insufficiency or increased admission creatinine, in-hospital worsening of renal function is an important risk factor for 6-month mortality in patients admitted with ACS. Furthermore, return to baseline function by discharge does not protect against this risk. These findings have implications for management of these high-risk patients. PMID- 17398189 TI - The editor's roundtable: intravascular ultrasonic imaging of the coronary arteries. PMID- 17398190 TI - Value of electrocardiographic and ankle-brachial index abnormalities for prediction of coronary atherosclerosis in asymptomatic subjects with type 2 diabetes mellitus. AB - Type 2 diabetes mellitus (DM) is associated with increased cardiovascular risk, in part due to accelerated subclinical atherosclerosis. Electrocardiographic (ECG) and ankle-brachial index (ABI) abnormalities are used to screen for cardiovascular risk in the clinic. However, their capacity to identify patients with type 2 DM with nonobstructive subclinical atherosclerosis is unknown. Associations of ECG and ABI abnormalities with coronary artery calcium (CAC), a measure of coronary atherosclerosis, were examined using multivariable ordinal regression modeling in 589 asymptomatic patients with type 2 DM. Sensitivity, specificity, and positive and negative predictive values were determined. CAC was prevalent (44% CAC>00; 32% CAC>5th percentile score) despite normal electrocardiograms (64%) and ABIs (97%) in most subjects. Neither ECG nor ABI changes predicted CAC after adjusting for age, gender, and race. ECG abnormalities were neither sensitive nor specific for detection of CAC>100, >400, or>75th percentile (sensitivities 0.43, 0.45, and 0.34; specificities 0.69, 0.66, and 0.63, respectively). ABI abnormalities were not sensitive (0.03, 0.04, and 0.03) but had high specificity (0.98, 0.98, and 0.98). In subjects with normal electrocardiograms and ABIs, extensive CAC was remarkably prevalent (CAC>00 in 24%). In conclusion, ECG and ABI abnormalities failed to detect patients with subclinical coronary atherosclerosis and therefore may be of limited value in identifying many asymptomatic patients with type 2 DM at increased risk of cardiovascular disease. PMID- 17398192 TI - Usefulness of right ventricular tissue Doppler imaging to predict outcome in left ventricular heart failure independent of left ventricular diastolic function. AB - It is unknown whether right ventricular (RV) tissue Doppler (TD) predicts outcome in patients with left ventricular (LV) heart failure (HF) independently of contemporary echocardiographic Doppler variables of LV diastolic function. Comprehensive echocardiographic Doppler examination was performed before discharge in 107 patients hospitalized with LV HF. The primary end point was cardiac death or rehospitalization for HF. Follow-up was complete for 100 of 107 patients a mean of 527 days after hospital discharge. There were no significant differences in baseline clinical variables (mean age 58+/-12 years, 46% women, 77% hypertensive, 48% diabetic, 41% current smokers, and 23% known coronary artery disease) in prediction of the primary end point. Compared with patients without an event, patients with an event had a larger left atrial volume index (42+/-16 vs 33+/-13 ml/m2, p=0.001), lower LV ejection fraction (35+/-19% vs 46+/ 22%, p=0.01), higher mitral peak early diastolic flow velocity/TD early diastolic velocity (19+/-7 vs 14+/-7, p=0.001), lower RV fractional area change (39+/-11% vs 43+/-10%, p=0.04), and lower RV TD systolic velocity (8+/-2 vs 10+/-3 cm/s, p=0.005). On Cox proportional hazards multivariate analysis, left atrial volume index (p=0.01), mitral peak early diastolic flow velocity/TD early diastolic velocity (p=0.03), and RV TD systolic velocity (p=0.04) were independent predictors of outcome. Even when contemporary echocardiographic Doppler measures of LV diastolic function are considered, RV TD systolic velocity is an independent predictor of cardiac death or rehospitalization for HF in patients hospitalized with HF and appears to be superior to conventional 2-dimensional parameters of RV function. PMID- 17398191 TI - Effect of antihypertensive therapy on progression of carotid intima-media thickness in patients with type 2 diabetes mellitus. AB - Antihypertensive therapy reduces the incidence of cardiovascular events in people with diabetes mellitus (DM). However, the effects of treatment on subclinical atherosclerosis are less well studied. Results of a post hoc cohort analysis of the association between antihypertensive treatment and carotid artery intima media thickness (CIMT) in the Troglitazone Atherosclerosis Regression Trial (TART), a randomized trial designed to evaluate the impact of troglitazone treatment on CIMT progression in adults with insulin-requiring type 2 DM, are reported. CIMT, blood pressure (BP), and use of antihypertensive agents were measured throughout the 2-year treatment period. In multivariable mixed-effects models, the annual rate of change in CIMT in relation to BP and duration of antihypertensive agent use, adjusting for covariates, was evaluated. Higher systolic BP was associated with a higher CIMT progression rate (p=0.03). This association was reduced by antihypertensive treatment in a duration-dependent manner (p for interaction=0.035). Adjustment for age, treatment assignment, and change in fasting glucose during the trial did not attenuate these associations. In conclusion, regular use of antihypertensive agents reduces the harmful impact of increased BP on atherosclerosis progression in patients with DM. The antiatherogenic effect of antihypertensive agents, including BP normalization and possible direct vascular wall protection, can be detected by CIMT progression using B-mode ultrasound in patients with DM. PMID- 17398193 TI - Qualitative observation of left ventricular multiphasic septal motion and septal to-lateral apical shuffle predicts left ventricular reverse remodeling after cardiac resynchronization therapy. AB - A multiphasic septal motion and typical septal-to-lateral apical shuffle of the left ventricle can be observed echocardiographically in some patients with left branch bundle block. The relation of both with left ventricular (LV) dyssynchrony according to tissue Doppler and LV reverse remodeling after cardiac resynchronization therapy was investigated. Fifty-three patients (37 men; age 68+/-8 years) with ischemic (n=26) or idiopathic (n=27) cardiomyopathy, baseline QRS duration 171+/-30 ms, LV ejection fraction 21+/-7%, and LV end-diastolic volume 257+/-91 ml were studied. LV dyssynchrony using tissue Doppler was considered present if the SD of the interval between QRS and onset of systolic velocity of 6 basal LV segments was >20 ms. Shuffle was evaluated visually independently by 5 cardiologists and considered present if observed in>or=1 view. LV reverse remodeling, defined as LV end-systolic volume decrease>or=10%, was observed in 37 patients (70%) after 3 months of CRT. Sensitivity and specificity of either shuffle or multiphasic septal motion for all 5 observers (range 90% to 97% and 67% to 83%, respectively) were found to predict LV dyssynchrony. To predict LV reverse remodeling, sensitivity and specificity from 87% to 92% and 69% to 81% were observed, respectively. In conclusion, the qualitative observation of a typical shuffle or multiphasic septal motion predicts LV dyssynchrony and LV reverse remodeling adequately. PMID- 17398194 TI - Design and baseline characteristics of the simvastatin and ezetimibe in aortic stenosis (SEAS) study. AB - Aortic valve stenosis and atherosclerotic disease have several risk factors in common, in particular, hypercholesterolemia. Histologically, the diseased valves appear to have areas of inflammation much like atherosclerotic plaques. The effect of lipid-lowering therapy on the progression of aortic stenosis (AS) is unclear, and there are no randomized treatment trials evaluating cardiovascular morbidity and mortality in such patients. The Simvastatin and Ezetimibe in Aortic Stenosis (SEAS) Study is a randomized, double-blind, placebo-controlled, multicenter study of a minimum 4 years' duration investigating the effect of lipid lowering with ezetimibe/simvastatin 10/40 mg/day in patients with asymptomatic AS with peak transvalvular jet velocity 2.5 to 4.0 m/s. Primary efficacy variables include aortic valve surgery and ischemic vascular events, including cardiovascular mortality, and second, the effect on echocardiographically evaluated progression of AS. The SEAS Study randomly assigned 1,873 patients (age 68+/-10 years, 39% women, mean transaortic maximum velocity 3.1+/-0.5 m/s) from 173 sites. Other baseline characteristics were mean blood pressure of 145+/-20/82+/-10 mm Hg (51% hypertensive); 55% were current or previous smokers; and most were overweight (mean body mass index 26.9 kg/m2). At baseline, mean total cholesterol was 5.7+/-1.0 mmol/L (222 mg/dl), low-density lipoprotein cholesterol was 3.6+/-0.9 mmol/L (139 mg/dl), high-density lipoprotein cholesterol was 1.5+/-0.4 mmol/L (58 mg/dl), and triglycerides were 1.4+/-0.7 mmol/L (126 mg/dl). The SEAS Study is the largest randomized trial to date in patients with AS and will allow determination of the prognostic value of aggressive lipid lowering in such patients. PMID- 17398195 TI - Acute cardiac functional and morphological changes after Anthracycline infusions in children. AB - The aim of this study was to describe the acute effects of anthracyclines on left ventricular systolic and diastolic function using different echocardiographic modalities. Thirteen children scheduled to receive anthracyclines were prospectively studied. They underwent complete 2-dimensional and Doppler echocardiographic evaluations, including tissue Doppler imaging, before the first dose and<2 hours after each of the first 3 doses of anthracyclines (dose range 30 to 75 mg/m2). After the first dose, increased end-diastolic wall thickness, decreased wall thickening, and a prolonged myocardial performance index were noted. Parameters of diastolic function changed significantly, with a lower mitral E wave, a decreased E/A ratio, and prolonged isovolumic relaxation time. Also, reduced longitudinal early diastolic myocardial velocity and myocardial velocity acceleration during isovolumic contraction as well as reduced peak longitudinal and radial systolic strain rate and strain were noted. All these parameters remained significantly lower after subsequent doses. After the second dose, significant changes in the shortening fraction and the ejection fraction compared with baseline became apparent. After the third dose, further deterioration in radial peak systolic strain was seen. In conclusion, low to moderate doses of anthracyclines acutely induce cardiac diastolic and systolic dysfunction. PMID- 17398196 TI - Progressive dilation of the ascending aorta in children with isolated bicuspid aortic valve. AB - Although patients with bicuspid aortic valves (BAVs) are predisposed to ascending aortic (AA) dilation, stenosis, and dissection, the development of aortic disease in children with BAVs is poorly described. The purposes of this study were to determine the rate of change of AA diameter in children with BAVs and to identify risk factors for the development of aortic dilation. The echocardiograms of 276 children aged<19 years (mean 8.5+/-5.3) with isolated BAVs were reviewed. Aortic measurements were normalized to z scores on the basis of body surface area. In a subset of 112 patients with serial examinations, aortic growth rates were calculated and risk factors for more rapid aortic growth determined. At presentation, 33 patients (12%) demonstrated marked AA dilation (z>4), and 70 (25%) were moderately abnormal (z between 2 and 4). The mean+/-SD AA diameter increased more than expected, at a rate of 0.18+/-0.30 z score per year (p<0.0001). In 61 patients with normal AA diameters on initial study, 22 (36%) had abnormal diameters, with z scores>2, at follow-up. Univariate analysis demonstrated right-noncoronary commissural fusion (p<0.02) and aortic valve gradient on initial examination (p<0.02) as significant predictors of AA growth. In multivariate analysis, however, the significance of gradient and valve morphology was diminished (p = 0.06 for both). In conclusion, the progression of AA diameter in patients with normal z scores on initial examination suggests that serial echocardiograms are required to screen for the development of significant aortic dilation. PMID- 17398197 TI - Transcatheter closure of perimembranous ventricular septal defects in infants and children using the Amplatzer perimembranous ventricular septal defect occluder. AB - There are very few published reports of the transcatheter closure of perimembranous ventricular septal defects (PMVSDs) using the Amplatzer PMVSD occluder with encouraging initial results. This report presents initial and 1 year results from 54 patients with PMVSDs who underwent transcatheter closure at 5 different institutions with the Amplatzer PMVSD occluder. Sixty-five patients with PMVSDs were enrolled at 5 European centers. Eleven of the 65 patients did not fulfill the patient selection criteria at the initial echocardiographic evaluation or at cardiac catheterization. As a result, a total of 54 patients underwent attempted transcatheter closure using the Amplatzer PMVSD occluder. The median age of the patients was 5.1+/-3.6 years (range 0.3 to 13), and the median weight 18.5+/-10.3 kg (range 5 to 45). Devices were permanently implanted in 49 of 54 patients. Complete occlusion of the communication at 1-year follow-up was observed in 46 of 49 patients (94%). Main early procedural complications included (1) device embolization (2 patients), (2) severe bradycardia with hemodynamic compromise (2 patients), and (3) Mobitz II (2:1) heart block (1 patient). Late procedural complications included complete heart block (1 patient). No other complications were observed during follow-up. In conclusion, the Amplatzer PMVSD occluder is promising device that can be used for transcatheter closure in selected patients with PMVSDs. Further studies and long-term follow-up are required before this technique enters routine clinical practice. PMID- 17398198 TI - Frequency and extent of media in the internal carotid artery in "endarterectomy" specimens. AB - Carotid endarterectomy has been performed now for approximately 50 years. No report, however, has described the presence of media in these "endarterectomy" specimens. From January 1997 through May 2003, 147 carotid endarterectomy specimens were examined histologically. Portions of arterial media were present in each of them. The maximal amount of circumferential media in any patient varied from>95% of the cross-sectional circumference in 55 patients (37%) to75% of the arterial circumference of the internal carotid artery contained media. The proper term for this operative procedure might better be "endomediaectomy" rather than "endarterectomy." PMID- 17398199 TI - Assessment of fetal myocardial performance using myocardial deformation analysis. AB - Conventional techniques for the assessment of cardiac function on the basis of M mode or 2-dimensional modalities are technically difficult, load dependent, and provide information on global ventricular function only. Newer techniques, which analyze myocardial performance, such as tissue velocity, strain, and especially the less load dependent strain rate, may provide more appropriate information. Myocardial systolic and diastolic motion and performance were calculated using tissue velocity, strain, and strain rate imaging on a large cohort of normal fetuses. The assessment of myocardial performance was feasible in all 98 normal fetuses. Normal systolic and diastolic values for tissue velocity, strain, and strain rate were established. All data were highly reproducible. Tissue velocity was age dependent, whereas strain and strain rate were stable throughout gestation. All parameters were heart rate independent. In conclusion, fetal myocardial velocity, strain, and strain rate measurements are easy to obtain and reproducible, and therefore, may serve as reference data. Increases in tissue velocity throughout gestation probably reflect the growth of the fetal heart, whereas intrinsic myocardial properties as measured by strain rate do not change. In comparison with recently published myocardial performance values in children, these strain rate data suggest that fetal myocontractile properties that are already established during the second half of pregnancy remain constant throughout gestation and after birth. PMID- 17398200 TI - Usefulness of an abnormal cardiovascular response during low-grade head-up tilt test for discriminating adolescents with chronic fatigue from healthy controls. AB - Hemodynamic dysfunction is documented in chronic fatigue syndrome (CFS). This study was conducted to investigate cardiovascular responses to orthostatic stress in adolescents with CFS, using a novel procedure for tilt-table testing. A total of 27 adolescents with CFS and 33 healthy control subjects with equal age and gender distribution underwent 15 minutes of 20 degrees head-up tilt testing. Heart rate, systolic blood pressure (BP), mean BP, diastolic BP, stroke index, total peripheral resistance index, end-diastolic volume index, and acceleration index were continuously and noninvasively recorded. At rest, patients with CFS had higher total peripheral resistance index values (p<0.01) and lower stroke index and end-diastolic volume index values (p<0.05) than controls. During 20 degrees head-up tilt testing, patients with CFS had greater increases in heart rate, diastolic BP (p<0.001), mean BP (p<0.01), and total peripheral resistance index (p<0.05) than controls and greater decreases in stroke index (p<0.05). Syncope or near syncope was not observed. In conclusion, this study found that adolescents with CFS have significant abnormalities of cardiovascular regulation in response to mild orthostatic stress, differentiating them from healthy controls. PMID- 17398201 TI - Impact of smart infusion technology on administration of anticoagulants (unfractionated Heparin, Argatroban, Lepirudin, and Bivalirudin). AB - This study reviewed 863 alerts generated from the infusion of anticoagulants in 355 patients from October 2003 to January 2005. Alerts were generated by smart infusion technology pumps and recorded in the devices' memory. The most common alerts were underdose alerts (59.8%), followed by overdose alerts (31.3%) and duplicate drug therapy alerts (8.9%). In response to the alerts, users' most frequent action was to cancel (46.5%) or reprogram (43.1%) the infusions. The highest percentage of alerts occurred from 2 to 4 p.m. During the study, there were 4 infusion rate errors, compared with 15 in the immediately preceding 16 month period. In conclusion, smart infusion technology intercepted keypad entry errors, thereby reducing the likelihood of intravenous anticoagulant overdose or underdose. Dose or infusion rate programming during intravenous anticoagulation is an important targets for medication safety interventions. PMID- 17398202 TI - Effect of inhibition of the renin-angiotensin system on development of type 2 diabetes mellitus (meta-analysis of randomized trials). AB - The increasing prevalence and costs of type 2 diabetes mellitus (DM) make strategies to prevent its development vitally important. This analysis was conducted to determine if angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs) prevent the development of DM. Medline and the Cochrane Central Register of Controlled Trials (1966 to May 2006) were queried for prospective, randomized, placebo-controlled or active-controlled trials of ACE inhibitor or ARB therapy in adults that reported rates of new-onset diabetes during follow-up. Meta-analyses of summary statistics from individual trials were performed using a random-effects model. Thirteen trials with a total of 93,451 patients were identified. Renin-angiotensin system antagonists reduced the incidence of DM from 9% in nontreated patients to 7.1% in those treated, a 26% reduction in odds (odds ratio [OR] 0.74, 95% confidence interval [CI] 0.66 to 0.81, p<0.001). The effect sizes were similar in trials that randomized only hypertensive subjects (OR 0.73, 95% CI 0.66 to 0.82, p<0.001) and trials that studied the impact of renin-angiotensin system inhibition on outcomes of patients with vascular disease or heart failure (OR 0.67, 95% CI 0.50 to 0.90, p=0.008). ACE inhibitors and ARBs had comparable effects on the development of DM. In ACE inhibitor trials, the odds of developing DM were reduced by 28% (OR 0.72, 95% CI 0.63 to 0.84, p<0.001), and in the 5 ARB studies, there was a 27% reduction (OR 0.73, 95% CI 0.64 to 0.84, p<0.001) in the odds. In conclusion, evidence accumulated to date indicates that inhibition of the renin-angiotensin system may contribute to the prevention of DM. PMID- 17398203 TI - The things to come of SHAPE: cost and effectiveness of cardiovascular prevention. AB - It has been proposed recently that asymptomatic adults undergo routine screening for subclinical atherosclerosis using electron-beam computed tomography or carotid ultrasound as the basis for the pharmacologic prevention of cardiovascular events. The expected cost and effectiveness of this conditional test-treatment strategy are herein analyzed in comparison with the conventional conditional strategy, based on Framingham risk factor assessment, and with an unconditional treatment strategy that does not rely on testing. These analyses show that the unconditional treatment strategy, albeit more costly, is more effective and more cost-effective than either conditional testing strategy. In conclusion, greater attention should be paid to improving the population-wide delivery of and long-term adherence to proven preventive therapy than to the identification of "high-risk" treatment targets. PMID- 17398204 TI - Assessing prognosis in patients with diabetes mellitus--the Achilles' heel of cardiac stress imaging tests? AB - The prognostic utility of stress imaging studies has been validated in numerous studies and, in general, patients with a normal imaging study have annual cardiac ischemic event rates of <1%. However, this predictive value of a normal stress imaging study does not appear to be applicable to subjects with diabetes. In this editorial, we summarize the current available data on prognostic utility of stress imaging studies in subjects with diabetes and provide insights into how to interpret and integrate these data for daily clinical practice. In conclusion, currently available data suggest that the prognostic value of stress imaging studies in subjects with diabetes differs from those without diabetes and should be interpreted as such. PMID- 17398205 TI - Late stent thrombosis: problem or not? PMID- 17398206 TI - Predictors of left ventricular remodeling after reperfused acute myocardial infarction. PMID- 17398207 TI - The triangle of valve calcification-cytokine-bone resorption: propose of novel potential pathophysiologic link. PMID- 17398208 TI - Selective serotonin reuptake inhibitors may interfere with the antiplatelet effect of clopidogrel. PMID- 17398209 TI - An unusual case of dysphagia and chest pain in a non-HIV patient: esophageal tuberculosis. PMID- 17398210 TI - Prehypertension and elevated risk of cardiovascular disease: physiopathologic mechanisms. PMID- 17398211 TI - The safety of levofloxacin in patients on warfarin. PMID- 17398213 TI - Porcelain gallbladder and cancer: ethnicity explains a discrepant literature? PMID- 17398214 TI - Dosing frequency of aspirin and prevention of heart attacks and strokes. PMID- 17398216 TI - Enteric coated aspirin. PMID- 17398217 TI - Peer review: the best of the blemished? PMID- 17398218 TI - Judicious antibiotic use and intranasal corticosteroids in acute rhinosinusitis. AB - Most patients with symptoms of acute rhinosinusitis are treated with antibiotics. However, many cases of rhinosinusitis are secondary to viral infections and unlikely to benefit from antibiotic therapy. Inappropriate use of antibiotics in patients with acute nonbacterial rhinosinusitis contributes to the increase in bacterial antibiotic resistance. Consequently, safe and effective alternatives to antibiotics are needed in the treatment of acute rhinosinusitis caused by viral infections. Recent results from controlled trials have shown that intranasal corticosteroids, used in combination with antibiotics or as monotherapy in selected cases, provide significant symptom relief and resolution of acute rhinosinusitis. The use of intranasal corticosteroids in acute rhinosinusitis therefore might reduce the inappropriate use of antimicrobial therapy in acute rhinosinusitis. PMID- 17398219 TI - Guideline for low-cost antimicrobial use in the outpatient setting. AB - In an effort to increase appropriate prescribing of low-cost antimicrobials in the outpatient setting, an evidence-based guideline was created to identify situations when low-cost medications can be used. A literature search identified relevant clinical trials describing the efficacy of antimicrobials used in the outpatient setting. These were analyzed to identify low-cost medications defined as $15 or less. The information was put into guideline format that includes the level of evidence for recommending the drug and information about cost. Sixteen common infections and their treatments were included in the guideline. The efficacy data were similar for the low-cost and higher-cost antimicrobials for all infections included. We created a low-cost antimicrobial guideline for common infections treated in the outpatient setting. The treatment options have similar efficacy to higher cost medications. This guideline will serve as an information source for providers to help them rapidly determine the low-cost treatments for common infections. In addition, it can serve as a template for the development of similar guidelines in other therapeutic classes. These guidelines should be customized before implementation at other health care organizations, with consideration of local resistance patterns, drug availability and patient factors. The effect of guideline implementation on future prescribing habits and providers' opinions about availability of cost information and subsequent conversations with patients and prescribers of medications deserves further study. PMID- 17398220 TI - Contemporary empyema necessitatis. AB - Empyema necessitatis is defined by the extension of an empyema through the parietal pleura, into surrounding tissue. Clinical manifestations are generally subacute, representing the indolent nature of the most commonly implicated pathogens (Mycobacterium tuberculosis and Actinomyces israelii). Treatment with antimicrobials and surgical debridement has drastically reduced mortality; however, with proper initial assessment and treatment of pulmonary infection, this rare complication can often be avoided. We describe a patient with empyema necessitatis to illustrate the importance of timely diagnosis and treatment of parapneumonic pleural effusion, and the need to consider this diagnosis in patients presenting with constitutional symptoms and a chest wall mass. PMID- 17398221 TI - Office management of chronic pain in the elderly. AB - Chronic pain plagues older adults more than any other age group; thus, practitioners must be able to approach this problem with confidence and skill. This article reviews the assessment and treatment of the most common chronic nonmalignant pain conditions that affect older adults--myofascial pain, generalized osteoarthritis, chronic low back pain (CLBP), fibromyalgia syndrome, and peripheral neuropathy. Specific topics include essential components of the physical examination; how and when to use basic and advanced imaging in older adults with CLBP; a stepped care approach to treating older adults with generalized osteoarthritis and CLBP, including noninvasive and invasive management techniques; how to diagnose and treat myofascial pain; strategies to identify the older adult with fibromyalgia syndrome and avoid unnecessary "diagnostic" testing; pharmacological treatment for the older adult with peripheral neuropathy; identification and treatment of other factors such as dementia and depression that may significantly influence response to pain treatment; and when to refer the patient to a pain specialist. While common, chronic pain is not a normal part of aging, and it should be treated with an emphasis on improved physical function and quality of life. PMID- 17398222 TI - Too much of a good thing. PMID- 17398223 TI - A case for vaccination. PMID- 17398224 TI - Mystery of the silk road. PMID- 17398225 TI - A racing heart. PMID- 17398226 TI - A complication of forceful nose-blowing. PMID- 17398227 TI - Vitamin and micronutrient intake and the risk of community-acquired pneumonia in US women. AB - BACKGROUND: There are limited data regarding the role of dietary and supplemental vitamin intake and the risk of community-acquired pneumonia. METHODS: We prospectively examined, during a 10-year period, the association between dietary and supplemental vitamin intake and the risk of community-acquired pneumonia among 83,165 women in Nurses' Health Study II who were between the ages of 27 and 44 years in 1991. We excluded women who had pneumonia before 1991, those who did not provide complete dietary information, or those with a history of cancer, cardiovascular disease, or asthma. Self-administered food frequency questionnaires were used to assess dietary and supplemental vitamin intake. Cases of pneumonia required a diagnosis by a physician and confirmation with a chest radiograph. The independent associations between specific vitamins and pneumonia risk were evaluated. RESULTS: There were 925 new cases of community-acquired pneumonia during 650,377 person-years of follow up. After adjusting for age, cigarette smoking, body mass index, physical activity, total energy intake, and alcohol consumption, there were no associations between dietary or total intake of any individual vitamin and risk of community-acquired pneumonia. Specifically, women in the highest quintile of vitamin A intake did not have a significantly lower risk of pneumonia than women in the lowest quintile (multivariate relative risk [RR]=0.88; 95% confidence interval [CI], 0.70-1.09, P for trend=.16). Similarly, vitamin C (RR=0.94; 95% CI, 0.76-1.16, P for trend=.81) and E (RR=0.95; 95% CI, 0.76-1.17, P for trend=.74) intake did not alter risk of pneumonia. CONCLUSIONS: Higher vitamin intake from diet and supplements is unlikely to reduce pneumonia risk in well nourished women. PMID- 17398228 TI - Grip strength predicts cause-specific mortality in middle-aged and elderly persons. AB - PURPOSE: Handgrip strength is a simple measurement used to estimate overall muscle strength but might also serve as a predictor of health-related prognosis. We investigated grip strength-mortality association in a longitudinal study. METHODS: A total of 4912 persons (1695 men and 3217 women), 35 to 74 years old at baseline, were the subjects of this study. Members of the Adult Health Study (AHS) cohort in Hiroshima, Japan, these individuals underwent a battery of physiological tests, including handgrip-strength testing, between July 1970 and June 1972. Mortality was followed until the end of 1999. Estimates of relative risk (RR) of mortality associated with grip strength were adjusted for potentially confounding factors by Cox proportional hazard analysis. RESULTS: Multivariate-adjusted RR of all causes of death, except for external causes, for the highest quintile of grip strength in men was 0.52 (95% confidence interval [CI], 0.33-0.80) for the age group 35-54 years, 0.72 (95% CI, 0.53-0.98) for the ages 55-64 years, and 0.67 (95% CI, 0.49-0.91) for the ages 65-74 years. These figures were significantly lower than the RR for the reference group (the third quintile). Similar trends were observed in women. Multivariate-adjusted RR of all causes of death except external causes for each 5-kg increment of grip strength was significantly low (RR: 0.89, 95% CI, 0.86-0.92 for men, RR: 0.87, 95% CI, 0.83-0.92 for women). Multivariate-adjusted RR for heart disease, stroke, and pneumonia in men was 0.85 (95% CI, 0.79-0.93), 0.90 (95% CI, 0.83-0.99), and 0.85 (95% CI, 0.75-0.98), respectively. RR for each 5-kg increment of grip strength remained 0.92 (95% CI, 0.87-0.96), even after more than 20 years of follow-up. CONCLUSION: Grip strength is an accurate and consistent predictor of all causes of mortality in middle-aged and elderly persons. PMID- 17398229 TI - Thyroid hormone use, hyperthyroidism and mortality in older women. AB - PURPOSE: Thyroid dysfunction is common, particularly among older women. The safety of thyroid hormone use and long-term prognosis of hyperthyroidism remain controversial. We performed a prospective cohort study to examine the relationship among thyroid hormone use, previous hyperthyroidism, abnormal thyroid function, and mortality. METHODS: We studied 9449 community-dwelling white women aged > or =65 years followed for 12 years. For analyses of thyroid function, we performed a nested case-cohort in 487 women using a third-generation thyroid-stimulating hormone assay. Causes of death were adjudicated based on death certificates and hospital records. RESULTS: Twelve percent of the 9449 women took thyroid hormone at baseline, and the mean duration of thyroid hormone use was 15.8 years; 9.4% of participants reported a history of hyperthyroidism. During 12 years of follow-up, 3159 women died (33%). In multivariate analysis, mortality among users of thyroid hormone was similar to that observed for nonusers (relative hazard [RH] 1.11, 95% confidence interval [CI], 0.98-1.24, P=.09). Previous hyperthyroidism was associated with a higher risk of all-cause mortality (RH 1.20, 95% CI, 1.06-1.36), particularly cardiovascular mortality (RH 1.46, 95% CI, 1.20-1.77). Low (< or /=0.5 mU/L) or high (>5 mU/L) thyroid stimulating hormone levels were not associated with excess total or cause specific mortality, but the power to detect these relationships was limited. CONCLUSIONS: Among older women, thyroid hormone use is not associated significantly with excess mortality, but previous hyperthyroidism may be associated with a small increase in all-cause and cardiovascular mortality. Additional long-term studies of hyperthyroidism and its treatment should further explore these findings. PMID- 17398230 TI - TB in a low-incidence country: differences between new immigrants, foreign-born residents and native residents. AB - BACKGROUND: New immigrants and foreign-born residents add to the burden of pulmonary tuberculosis (TB) in low-incidence countries. The highest TB rates have been found among recent immigrants. Active screening programs are likely to change the clinical presentation of TB, but the extent of the difference between immigrant and resident populations has not been studied prospectively. METHODS: Adult new immigrants were screened upon entry to 1 of 5 immigration centers in Switzerland. Immigrants with abnormal chest radiographs were enrolled and compared in a cohort study to consecutive admitted foreign-born residents from moderate-to-high incidence countries and native residents presenting with suspected TB. RESULTS: Of 42,601 new immigrants screened, 112 had chest radiographs suspicious for TB. They were compared with foreign-born residents (n=118) and native residents (n=155) with suspected TB (n=385 patients included). Active TB was confirmed in 40.5% of all patients (immigrants 38.4%, foreign-born residents 50%, native residents 34.8%). Clinical signs and symptoms of TB and laboratory markers of inflammation were significantly less common in immigrants than in the other groups with normal results in >70%. The proportion of positive results on rapid testing to detect M. tuberculosis (MTB) in 3 respiratory specimens was significantly lower in immigrants (34.9% for acid-fast staining; 55.8% for polymerase chain reaction) compared with foreign-born residents (76.2% and 89.1%, respectively) and native residents (83.3% and 90.9%, respectively). Isoniazid resistance and multi-drug resistance were more prevalent in immigrants. CONCLUSION: New immigrants with TB detected in a screening program are often asymptomatic and have a low yield of rapid diagnostic tests but are at higher risk for resistant MTB strains. Postmigration follow-up of pulmonary infiltrates is essential in order to control TB among immigrants, even in the absence of clinical and laboratory signs of infection. PMID- 17398231 TI - Coronary ischemic events after first atrial fibrillation: risk and survival. AB - PURPOSE: We aimed to determine the long-term, gender-specific incidence and mortality risk of coronary ischemic events after first atrial fibrillation (AF). METHODS: In this longitudinal cohort study, adult residents of Olmsted County, Minnesota, with an electrocardiogram-confirmed AF first documented in 1980 to 2000 and without prior coronary heart disease, were followed to 2004. The primary outcome was first coronary events (angina with angiographic confirmation, unstable angina, nonfatal myocardial infarction, or coronary death). Sex-specific incidence of coronary ischemic events and survival after development of such events were assessed using Cox proportional hazards modeling. Kaplan-Meier estimates of risks for coronary ischemic events were compared with those predicted by the Framingham equation. RESULTS: Of the 2768 subjects (mean age 71 years, 48% were men), 463 (17%) had a first coronary event during a follow-up of 6.0+/-5.2 years. The unadjusted incidence was 31 per 1000 person-years, and there was no difference between men and women. The incidence was higher in men (hazard ratio 1.32, P=.004) after adjusting for age. The 10-year event estimates were 22% and 19% in men and women, respectively, by our Kaplan-Meier analyses, and 21% and 11%, respectively, by Framingham risk equation. The mortality risk after coronary events was higher in women (hazard ratio 2.99 vs 2.33; P=.044), even after multiple adjustment. CONCLUSIONS: First AF marks a high risk for new coronary ischemic events in both men and women. AF conferred additional risk for coronary events beyond conventional risk prediction in women only. The excess mortality risk associated with the development of coronary events was significantly greater in women. PMID- 17398232 TI - Outbreak of pertussis on a college campus. AB - BACKGROUND: Pertussis is increasing among adolescents and adults despite universal childhood vaccination. This investigation describes an outbreak of pertussis among undergraduate students and assesses the burden of cough illness on a college campus. METHODS: Students presenting with prolonged cough were evaluated with culture, polymerase chain reaction (PCR), and serology. An e-mail survey was performed to determine the burden of cough illness on campus. RESULTS: Thirty-seven undergraduates were evaluated. Their mean duration of cough was 28 days. No student had cultures positive for B. pertussis; one was PCR positive. Ten (27%) had serologic values consistent with acute pertussis infection. The e mail survey was returned by 225/500 (45%) students. Of these, 66 (29%; 95% confidence interval [CI], 23%-36%) reported a cough of 2 weeks or longer duration during the fall semester. A conservative estimate showed that the campus-wide incidence of a cough illness meeting the Centers for Disease Control and Prevention case definition for pertussis was 13% (95% CI, 10%-16%) during the fall semester. CONCLUSIONS: Adolescents and young adults are susceptible to pertussis infection. This study demonstrates that there was a substantial rate of pertussis infection during an outbreak on a college campus. Our findings support the routine use of the acellular pertussis vaccine in adolescents and adults. PMID- 17398233 TI - Prognostic stratification of patients with left-sided endocarditis determined at admission. AB - BACKGROUND: The prognosis of patients with left-sided endocarditis remains poor despite the progress of surgical techniques. Identification of high-risk patients within the first days after admission to the hospital would permit a more aggressive therapeutic approach. METHODS: We designed a prospective multicenter study to find out the clinical, microbiologic, and echocardiographic characteristics available within 72 hours of admission that might define the profile of high-risk patients. Of 444 episodes, 317 left-sided endocarditis cases were included and 76 variables were assessed. Events were surgery in the active phase of the disease and in-hospital death. A stepwise logistic regression analysis was undertaken to determine variables predictive of events. RESULTS: Multivariate analysis of the clinical variables found to have statistical significance in the univariate analysis identified the following as predictive: patient referred from another hospital (odds ratio [OR]: 1.8; confidence interval [CI], 1.1-2.9), atrioventricular block (OR: 2.5; CI, 1.1-5.9), acute onset (OR: 1.7; CI, 1.1-2.9), and heart failure at admission (OR: 2.3; CI, 1.4-3.8). When the echocardiographic and microbiological variables statistically significant in the univariate analysis were introduced, the presence of heart failure at admission (OR: 2.9; CI, 1.8-4.8), periannular complications (OR: 1.8; CI, 1.1 3.1), and Staphylococcus aureus infection (OR: 2.0; CI, 1.1-3.8) retained prognostic power. Risk could be accurately stratified when combining the 3 variables with predictive power: 0 variables present: 25% of risk; 1 variable present: 38% to 49% of risk; 2 variables present: 56% to 66% of risk; and 3 variables present: 79% of risk. CONCLUSIONS: The risk of patients with left-sided endocarditis can be accurately stratified with the assessment of variables easily available within 72 hours of admission to the hospital. PMID- 17398234 TI - Does statin therapy decrease the risk for bleeding in patients who are receiving warfarin? AB - PURPOSE: Recent observations in patients with atrial fibrillation who are receiving warfarin suggest that concomitant treatment with a 3-hydroxy-3 methylglutaryl coenzyme A reductase inhibitor (statin) decreases the risk for bleeding. METHODS: We conducted a population-based, nested case-control study using the linked administrative databases of Ontario, Canada, to assess whether statin use decreases the risk of bleeding in warfarin users. Eligible patients were Ontario residents, age 66 years or more, with atrial fibrillation who were prescribed warfarin between April 1, 1994, and December 31, 2001. Patients were followed until hospitalization for upper gastrointestinal or intracranial bleeding, study end (March 31, 2002), discontinuation of warfarin, or death. Cases were matched to controls by age and sex. Logistic regression analysis was used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the association between bleeding and statin use. RESULTS: We identified 79,207 warfarin users with atrial fibrillation. There were 1518 cases with an upper gastrointestinal or intracranial bleed and 15,100 matched controls without bleeding. Long-term (>/=1 year) statin use was associated with a lower risk for any bleeding (OR=0.80; 95% CI, 0.66-0.97). However, there was no association between bleeding and recent (<6 months) statin use (OR=1.04; 95% CI, 0.74-1.48) or statin use of any duration (OR: 0.91; 95% CI, 0.77-1.07), suggesting potential confounding of the association between statin use and bleeding by a health-user effect. CONCLUSION: Long-term statin use may be associated with a decreased risk for bleeding in warfarin users with atrial fibrillation. Additional research is needed to further explore this putative association. PMID- 17398235 TI - Late diagnosis of HIV infection: the role of age and sex. AB - BACKGROUND: Late diagnosis of human immunodeficiency virus (HIV) infection is detrimental to infected persons and to the public health. The objective of this study was to identify factors associated with late diagnosis of HIV infection, defined as an initial CD4 count <200 cells/microL, in a cohort of recently diagnosed persons. Additionally, we evaluated factors associated with HIV infection being diagnosed during hospitalization. METHODS: This was a cross sectional study of a university-based HIV clinic in the southeastern US. Patients with newly diagnosed HIV infection evaluated at the Duke University HIV clinic between October 2002 and August 2004 were included in this analysis. Socio demographic variables, site of HIV diagnosis, opportunistic infections present at diagnosis, initial CD4 count, and initial HIV RNA level were recorded for study subjects. RESULTS: Forty-nine percent of subjects met the immunologic definition of AIDS at the time of HIV diagnosis (CD4 count <200 cells/microL). In multivariable logistic regression analyses, older patients were more likely to be diagnosed with a CD4 count <200 cells/microL (adjusted odds ratio [AOR] 1.72, 95% confidence interval [CI], 1.12-2.64, P=.01), and older patients (AOR 1.79, 95% CI, 1.07-3.12, P=.03) and women (AOR 6.74, 95% CI, 2.08-21.81, P=0.001) were more likely to be diagnosed during hospitalization. CONCLUSIONS: Late diagnosis of HIV infection is a considerable problem, particularly for older patients. Inpatient diagnosis of HIV infection is significantly more common among women and older patients. Improved HIV testing strategies may allow for more timely diagnosis of HIV infection, which may benefit both the infected individual and society. PMID- 17398236 TI - The lost art of clinical skills. PMID- 17398238 TI - American Congress of Rehabilitation Medicine in 2006: embracing the future. AB - The American Congress of Rehabilitation Medicine (ACRM) modified its mission and structure in 1997 to become an organization focused on medical rehabilitation research. Initially, this transformation accelerated an already diminishing membership, a weakened financial condition, and some level of dysfunction within the organizational structure. In recent years, with the advent of evidence-based practice and the expectation that empirical research is critical to the survival of clinical specialties such as rehabilitation medicine, ACRM has become re energized. New initiatives have been spawned that have led to stabilization and an influx of new members, a measurable improvement in the quality of scientific presentations at, and participation in, the annual meetings, efforts directed toward increasing the visibility and involvement of ACRM on an international level, programming directed toward early career scientists, strong public policy advocacy, and renewed and expanded inter-organizational partnerships. In addition, the financial position of ACRM has improved markedly and prospects toward long-term fiscal health and growth are "the new reality." ACRM has now moved significantly beyond the "survival" step of the Maslow hierarchy toward the goal of self-actualization. PMID- 17398239 TI - Through the looking glass: a personal view of the field of rehabilitation medicine. The 56th John Stanley Coulter Memorial Lecture. AB - This lectureship reviews the past, present, and future of the field of physical medicine and rehabilitation (PM&R) from the clinical, educational, and research points of view. I make recommendations for the field on how members of the various PM&R associations can make a difference in mapping the future. PMID- 17398240 TI - Satisfaction with quality of life poststroke: effect of sex differences in pain response. AB - OBJECTIVE: To study the relationship between sex differences among men and women in the response to pain and the effect on satisfaction with quality of life (QOL) in poststroke patients approximately 90 days after discharge from inpatient medical rehabilitation. DESIGN: Cross-sectional design. SETTING: Community based. PARTICIPANTS: The sample included 1724 patients (877 men, 847 women) aged 40 years and older with a history of stroke, according to information obtained from the IT HealthTrack database. The average age was 68.7 years; 79.3% were non Hispanic white. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Satisfaction with QOL. RESULTS: Pain ratings for both the men and the women were significant and inversely associated with satisfaction with QOL after adjustments for possible confounding factors. However, among men with stroke, each 1-point increase in pain rating was associated with an 18% decreased odds of being satisfied with QOL (odds ratio [OR], .81; 95% confidence interval [CI], .77-.86). Conversely, among women with stroke, each 1-point increase in pain rating was associated with an 11% decreased odds of being satisfied with QOL (OR=.89; 95% CI, .84-.95). CONCLUSIONS: Our data indicate that pain after stroke is significantly associated with reduced satisfaction with QOL, particularly among men. Increased awareness and treatment of pain may improve satisfaction with QOL. Our findings suggest that sex differences and pain response are clinically important factors in examining satisfaction with QOL in the poststroke population. PMID- 17398241 TI - Comparison of the Traumatic Brain Injury (TBI) Model Systems national dataset to a population-based cohort of TBI hospitalizations. AB - OBJECTIVE: To determine whether severity alone accounts for differences observed between a population-based cohort of acute care hospitalizations for traumatic brain injury (TBI) and the Traumatic Brain Injury Model Systems (TBIMS) national dataset. DESIGN: Prospective cohort. SETTING: Acute care hospitals in South Carolina and TBIMS rehabilitation centers. PARTICIPANTS: Subjects enrolled in the TBIMS national dataset and the South Carolina TBI Follow-up Registry (SCTBIFR). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Comparable variables in the 2 datasets included demographic characteristics, etiology of injury, initial Glasgow Coma Scale score, Abbreviated Injury Scale score for the head region derived from International Classification of Diseases codes, presence of computed tomography (CT) abnormalities, acute hospital length of stay, and payer source. RESULTS: As hypothesized, TBIMS participants showed greater initial injury severity, frequency of abnormal CT scans, and longer lengths of acute care hospitalization, explaining over 75% of cohort membership. Counter to a priori hypotheses, when all other factors were held constant, there were also differences in racial and ethnic background and insurance payer source. CONCLUSIONS: Differences between the TBIMS cohort and patients acutely hospitalized with TBI are primarily due to injury severity; however, an additional difference in payer source may need to be taken into account when generalizing findings. Results showed that TBIMS and SCTBIFR datasets are complementary, each having different strengths for understanding factors that impact long-term recovery after TBI. Recommendations are made for methodologic improvements in both data collection for the TBIMS and future outcome surveillance. PMID- 17398242 TI - Community participation after spinal cord injury. AB - OBJECTIVES: To describe participation among a community-based sample of adults with spinal cord injury (SCI) and to explore the relationship between participation and life satisfaction. DESIGN: Survey. SETTING: Community. PARTICIPANTS: A sample of 357 people (response rate, 44%) with SCI recruited through the British Columbia Paraplegic Association. The mean age +/- standard deviation was 46.0+/-14.7 years, mean time since SCI was 13.0+/-11.0 years, and 68% of the respondents were men. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Descriptive survey data on community participation specifically related to social involvement, physical activity and relationship with others, transportation, and community access. Life satisfaction and happiness assessed using the Satisfaction With Life Scale and Happiness Scale. RESULTS: No limitations to participation were experienced by 18.5% of the respondents. Satisfaction with transportation was associated with owning one's own vehicle (P<.001). There was overall satisfaction with access to community buildings (mean score range, 6.9-8.5; where 10 is most satisfied). Being physically active was important to a majority and 75% were currently engaged in physical activity. Those living alone were less satisfied (mean, 7.3+/-2.7; where 10 is most satisfied) with the support they received than those living with others (8.5+/ 1.7). Respondents were generally happy (5.0+/-1.4) and satisfied with life (18.6+/-7.6). CONCLUSIONS: This study provides a rich description of the multifaceted nature of participation and its association with life satisfaction as identified by people with SCI living in the community. PMID- 17398243 TI - Wheelchair use by veterans newly prescribed a manual wheelchair. AB - OBJECTIVE: To describe the characteristics of the wheelchairs, the users, and their wheelchair use among persons newly prescribed a manual wheelchair. DESIGN: Cohort study. SETTING: Veterans Affairs teaching hospital. PARTICIPANTS: Ninety nine consecutive, cognitively intact veterans prescribed a manual wheelchair. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Self-reported difficulty transferring into and propelling the wheelchair; and bathroom mobility method. RESULTS: Study patients had a mean age of 66 and a mean of 10 comorbid medical conditions. Parkinsonism, osteoporosis, joint replacement, and amputation were uncommon (<30% of patients), but had a high impact on need for a wheelchair (when present were reported by >50% of patients as causing need for a wheelchair). Falls and arthritis were common (>50% of patients) and highly impacted need for a wheelchair. At 1 month, over 30% of patients had wheelchairs that did not meet common criteria for wheelchair fit; 36% and 61%, respectively, reported difficulty transferring and propelling the wheelchair. The wheelchairs were used for bathroom mobility by 38% of the patients. CONCLUSIONS: The typical manual wheelchair recipient in this study sample was old with multiple medical problems. Despite provision of manual wheelchairs by trained professionals and availability of diverse wheelchair types, new wheelchair users commonly reported difficulty using the wheelchair. PMID- 17398244 TI - Rehabilitation outcomes of terror victims with multiple traumas. AB - OBJECTIVES: To describe the rehabilitation outcomes of terror victims with multiple traumas, and to compare those outcomes with those of patients with nonterror-related multiple traumas treated in the same rehabilitation facility over the same time period. DESIGN: Retrospective chart reviews. SETTING: Rehabilitation department in a university hospital in Jerusalem, Israel. PARTICIPANTS: Between September 2000 and September 2004, we treated 72 victims of terrorist attacks who had multiple traumas. Among them, 47 (65%) had multiple traumas without central nervous system involvement (MT subgroup), 19 (26%) had multiple traumas with traumatic brain injury (TBI subgroup), and 6 (8%) had multiple traumas with spinal cord injury (SCI subgroup). We matched, according to their types of injury and demographic data, each terror victim with a control patient treated in the same period in our rehabilitation department. INTERVENTION: Interdisciplinary inpatient and outpatient rehabilitation. MAIN OUTCOME MEASURES: Hospital length of stay (LOS) in acute care departments, inpatient and outpatient rehabilitation departments, functional outcome (FIM instrument score), occupational outcome (returning to previous occupation), and psychologic outcome (Solomon PTSD [post-traumatic stress disorder] Inventory). RESULTS: The mean LOS of terror victims was 218+/-131 days; for the nonterror group it was 152+/-114 days (P<.01). In comparison with the control subgroups, the MT subgroup of terrorist victims had significantly longer LOS in the acute care and outpatient rehabilitation departments (P=.06) and the terror TBI subgroup had a longer LOS in outpatient department only (P<.05). The LOS of the SCI patients, both terror victims and control patients, was significantly longer than that of the other 2 subgroups. The difference between FIM value at entry and discharge (DeltaFIM) was significantly higher for terror victims than for the controls (41.1+/-21.6 vs 30.8+/-21.8, P=.002). This difference was mainly the result of the significantly higher DeltaFIM achieved by the terror MT subgroup than by the MT controls. The rate of PTSD was higher among terror victims than among controls (40.9% vs 24.2%, P=.04). The rate of return to previous occupations was similar between terror victims and nonterror patients (53% vs 46.9%, respectively). CONCLUSIONS: Victims of terror spent longer periods in rehabilitation than the nonterror group; however, they regained most activity of daily living functions similar to the nonterror group. Despite the higher rate of PTSD, terror victims succeeded in returning to their previous occupations at a similar rate to that of the nonterror group. PMID- 17398245 TI - Prevalence of facet joint pain in chronic low back pain in postsurgical patients by controlled comparative local anesthetic blocks. AB - OBJECTIVE: To evaluate the prevalence of facet joint pain in patients with chronic low back pain (CLBP) after surgical intervention(s). DESIGN: A prospective, nonrandomized, consecutive study. SETTING: An ambulatory interventional pain management setting. PARTICIPANTS: The prevalence of facet joint pain was evaluated in patients with CLBP after various surgical intervention(s) referred to an interventional pain management practice. The sample was derived from 282 patients with persistent CLBP after various surgical intervention(s). Of these, 242 patients consented to undergo interventional techniques. A total of 117 consecutive patients with chronic, nonspecific low back pain, after lumbar surgical intervention(s) were evaluated with controlled, comparative local anesthetic blocks. INTERVENTIONS: Controlled, comparative local anesthetic blocks (1% lidocaine or 1% lidocaine followed by .25% bupivacaine) under fluoroscopic visualization using 0.5mL to block each facet joint nerve. MAIN OUTCOME MEASURES: A positive response was defined as at least 80% reduction of pain with ability to perform previously painful movements. A positive response was considered to be pain relief from the lidocaine block lasting at least 1 hour or at least 2 hours or greater than duration of relief with lidocaine when bupivacaine was used. Controlled, comparative local anesthetic blocks were used to eliminate false-positive results. Valid information is only obtained by performing controlled blocks in the form of comparative local anesthetic blocks, in which, on 2 separate occasions, the same joint is anesthetized by using local anesthetics with different durations of action. If patients obtained appropriate response with both blocks, they were considered a positive. If they obtained appropriate response with lidocaine but not with bupivacaine, they were considered false-positive, whereas if the response was negative with lidocaine, they were considered negative. RESULTS: The prevalence of lumbar facet joint pain in patients with recurrent pain after various surgical intervention(s) was 16% (95% confidence interval, 9%-23%). The false-positive rate with a single block with lidocaine was 49%. CONCLUSIONS: Facet joints are clinically important pain generators in a small but significant proportion of patients with recurrent CLBP after various surgical intervention(s). PMID- 17398246 TI - Upper-limb joint power and its distribution in spinal cord injured wheelchair users: steady-state self-selected speed versus maximal acceleration trials. AB - OBJECTIVE: To compare upper-limb joint power magnitude and distribution between the shoulder, elbow, and wrist during maximal acceleration (MAC) versus steady state, self-selected speed (SSS) manual wheelchair propulsion. DESIGN: Cross sectional biomechanic study. SETTING: Research university and teaching hospital. PARTICIPANTS: Volunteer sample of 13 manual wheelchair users with spinal cord injury below T1. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Propulsive joint power magnitude and fractional distribution among upper-limb joints. RESULTS: Wilcoxon signed-rank testing revealed shoulder power was larger for MAC versus SSS (median peak, 101.5W; interquartile range [IQR], 74.6; median peak, 37.7W; IQR, 22.9; respectively) (P<.01). Elbow and wrist power were unchanged. Peak shoulder power fraction was larger for MAC versus SSS (median peak, 1.055; IQR, .110 vs peak, .870; IQR, .252) (P<.01). Peak elbow power fraction was smaller for MAC versus SSS (median peak, -.012; IQR, .144 vs peak, .146; IQR, .206) (P<.05). Peak wrist power fraction was smaller for MAC versus SSS (median peak, -.058; IQR, .057 vs peak, -.010; IQR, .150) (P<.05). CONCLUSIONS: Power at the shoulder was larger than at other joints. Peak shoulder joint power and power fraction was larger during MAC versus SSS propulsion. Elbow and wrist power fractions were smaller for MAC versus SSS propulsion. Higher joint power, present under MAC, may predispose manual wheelchair users to injury, particularly at the shoulder. PMID- 17398247 TI - Preliminary evaluation of an automatically stance-phase controlled pediatric prosthetic knee joint using quantitative gait analysis. AB - OBJECTIVES: To evaluate the effects on gait of a new pediatric prosthetic knee joint using an automatic stance-phase lock (ASPL), shown previously to help prevent falls, and to propose future design considerations and prosthetic alignments. DESIGN: Case series and crossover trial. SETTING: Human movement laboratory. PARTICIPANTS: Three children with unilateral above-knee amputations and 3 children with bilateral above-knee or below-knee amputations. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Spatiotemporal, kinematic, and kinetic gait parameters. RESULTS: Spatiotemporal parameters indicated higher gait velocities with the ASPL knee joint for the children with unilateral amputations. The increased speed, as expected, was associated with increased temporal interlimb asymmetry, joint moments and powers, and excessive prosthetic knee range of motion in swing. A trend toward increased pelvic motions was observed with ASPL knee when compared with conventional knees. CONCLUSIONS: The biomechanic performance of the single-axis ASPL knee joint was shown to be comparable with more complex polycentric pediatric prosthetic knee joint technologies worn by the children in this study. PMID- 17398248 TI - Acute peripheral blood flow response induced by passive leg cycle exercise in people with spinal cord injury. AB - OBJECTIVE: To determine the acute femoral artery hemodynamic response in paraplegic subjects during a passive leg cycle exercise. DESIGN: Case series. SETTING: Department of physical medicine and rehabilitation in a university in France. PARTICIPANTS: A volunteer sample of 15 people with traumatic spinal cord injury. INTERVENTION: Subjects performed a 10-minute session of passive leg cycle exercise in the sitting position. MAIN OUTCOME MEASURES: We measured heart rate, maximal (Vmax), and minimal femoral artery blood flow velocity at rest and immediately after the passive leg cycle exercise, using quantitative duplex Doppler ultrasound. We calculated mean blood flow velocity (Vmean) and velocity index, representing the peripheral resistance, for each condition. RESULTS: Vmax and Vmean increased (from .80+/-.18 m/s to .96+/-.24 m/s, P<.01; and from .058+/ .02 m/s to .076+/-.03 m/s, P<.01; respectively) after 10 minutes of passive leg cycle exercise. Heart rate did not change. The velocity index decreased from 1.23+/-0.15 to 1.16+/-0.21 (P=.038). CONCLUSIONS: The results of this study suggest that acute passive leg cycle exercise increases vascular blood flow velocity in paralyzed legs of people with paraplegia. This exercise could have clinical implications for immobilized persons. PMID- 17398249 TI - Comparing saphenous nerve conduction study techniques at the knee and at the ankle and their relationship to body mass index. AB - OBJECTIVE: To evaluate the saphenous sensory response by 2 methods and the correlation of obtaining responses bilaterally with body mass index (BMI). DESIGN: Twenty patients evaluated by 1 electromyographer and 10 patients evaluated by 2 blinded electromyographers. SETTING: University electromyography laboratory. PARTICIPANTS: Thirty healthy adult volunteers. INTERVENTIONS: Saphenous sensory response with the recording electrode at the level of the medial malleolus, saphenous sensory response with the recording electrode at the level of the proximal tibia, and sural sensory response with the recording electrode posterior to the lateral malleolus. MAIN OUTCOME MEASURES: Sensory evoked amplitude and distal latency. RESULTS: The saphenous sensory response was obtained bilaterally at the knee in 77% of subjects, whereas it was obtained bilaterally at the ankle in only 50% of subjects (P<.05). BMI of greater than 25 kg/m2 resulted in more difficulty in obtaining the saphenous sensory response at the ankle compared with the response at the knee (P<.05). CONCLUSIONS: The saphenous sensory response at the knee is more consistently obtained than the saphenous sensory response at the ankle, and it is not affected by BMI. PMID- 17398250 TI - Development and psychometric properties of the Self-Efficacy Scale for transfers for caregivers of children requiring transfer assistance. AB - OBJECTIVE: To test the psychometric properties of a newly developed measure of caregiver self-efficacy, the Caregiver Self-Efficacy Scale for Transfers (CSEST), a 14-item questionnaire that measures the perceived self-efficacy of adult caregivers of dependent children and focuses on their perceived abilities and the challenges of assisting a child with restricted transfer ability. DESIGN: Reliability and validity study using survey data. SETTING: Community-based survey study. PARTICIPANTS: Adult caregivers (eg, mother, father, grandparent) of dependent minors who require transfer assistance. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Using the Rasch measurement model, the CSEST was tested for reliability and separation, validity (hierarchical and content), and factor structure. RESULTS: The CSEST yields data with sufficient reliability (reliability indices of .96 and .94 for respondents and items, respectively) as well as sufficient separation of levels of self-efficacy (item separation index, 4.13; person separation index, 5.23). The CSEST has a stable hierarchical structure with adequate content validity (standardized chi2 fit statistics <2.0) and point biserial correlations greater than .67. Factor analysis confirmed 2 insubstantial subscales; the 14-item CSEST accounted for greater than 85% of the total variance. CONCLUSIONS: The CSEST yields reliable data and valid inferences of self-efficacy from adult caregivers of children requiring transfer assistance. The CSEST may identify areas of need and/or difficulty with transfers for parents caring for dependent children. PMID- 17398252 TI - Validation of the Chinese translated Activities-Specific Balance Confidence scale. AB - OBJECTIVES: To translate contents of the Activities-Specific Balance Confidence (ABC) scale into Chinese (Cantonese), to examine the psychometric properties (internal reliability and validity) of this scale for use with Chinese older adults in Hong Kong, and to determine the level of self-perceived balance confidence of this population. DESIGN: A 2-phase exploratory study: formative qualitative research and a cross-sectional survey to establish the psychometric properties of the Chinese translated ABC (ABC-C) scale. SETTING: Community based. PARTICIPANTS: One hundred older community-dwelling subjects. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Measurement of perceived level of confidence in maintaining balance using the ABC-C scale. This instrument contains 16 items on indoor and outdoor activities requiring different levels of balance function. RESULTS: Four of the 16 ABC-C scale items had to be modified to achieve content equivalence and cultural relevance to Hong Kong. Final test items of this scale showed high internal consistency with a Cronbach alpha coefficient of .97. The instrument was found to have excellent test-retest and very good interrater reliability with intraclass coefficients of .99 and .85, respectively. Results of a factor analysis conducted on the scale showed evidence of a coherent 1-domain structure. The mean perceived balance confidence level +/- standard deviation of 100 community-dwelling older adults in Hong Kong, performed by using the ABC-C scale, was 71.6+/-23.7. CONCLUSIONS: Results of the present study verified that the ABC-C scale is a culturally relevant, valid, and reliable tool for measuring self-perceived balance confidence in Chinese older adults. These were reflected in very good to excellent internal consistency of items, test-retest and interrater reliability, and construct validity shown. PMID- 17398251 TI - Reliability and validity of the six-minute arm test for the evaluation of cardiovascular fitness in people with spinal cord injury. AB - OBJECTIVES: To design a submaximal arm ergometry test (six-minute arm test [6 MAT]), for persons with spinal cord injury (SCI) and to determine the test-retest reliability and concurrent validity of this test. DESIGN: Prospective, exploratory, methodologic study. To determine test-retest reliability, subjects completed the 6-MAT on 2 days, separated by 1 week. Validity was determined by comparing 6-MAT results with peak oxygen consumption (VO2peak). SETTING: Tertiary rehabilitation center. PARTICIPANTS: Thirty subjects with SCI (mean age, 36.3y; 83% male). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Subjects were evaluated on the 6-MAT and a VO2peak test. RESULTS: All subjects were able to complete the 6-MAT. Test-retest reliability of steady-state oxygen consumption (VO2) and heart rate during the 6-MAT were excellent (intraclass correlation coefficient [ICC], .81; 95% confidence interval [CI], .58-.92; ICC=.90; 95% CI, .75-.96, respectively). The correlation between VO2peak and 6-MAT VO2 was excellent (r=.92) and the correlations between VO2peak and 6-MAT heart rate (r=.63) and VO2peak and 6-MAT power output (r=.73) were good. CONCLUSIONS: This study showed that the 6-MAT has acceptable values for test-retest reliability and validity. The 6-MAT should be further tested for responsiveness to enhance its use as a clinical tool. PMID- 17398253 TI - Ulnar neuropathy at or distal to the wrist: traumatic versus cumulative stress cases. AB - OBJECTIVE: To identify clinical and electromyographic characteristics of ulnar neuropathy at or below the wrist, comparing those caused by unitary trauma with those caused by suspected cumulative stress. DESIGN: Retrospective case series. SETTING: University hospital electromyography laboratory. PARTICIPANTS: Patients with electrodiagnostic evidence of an ulnar neuropathy at or distal to the wrist over a 3-year period. Forty-seven hands from 42 patients (age range, 20-80y; mean, 52y) were identified and evaluated in this study. INTERVENTIONS: Record review of clinical history, physical examination, electromyography, and treatment. MAIN OUTCOME MEASURES: Etiology of injury, physical signs and symptoms, and electromyographic testing results. RESULTS: Ulnar neuropathy at or distal to the wrist is commonly mischaracterized because of other mononeuropathies in the upper extremity and because of peripheral polyneuropathy. Ulnar neuropathy because of cumulative stress presents typically with sensory symptoms (63%) and a normal examination (71%), whereas trauma cases present with motor with or without sensory symptoms (92%) with motor abnormalities (92%) confirmed on examination. Traumatic cases are characterized by electromyography by decreased sensory and motor-evoked amplitudes, prolonged motor distal latencies, and abnormal needle examination. The amplitude changes are noted comparing with laboratory norms and comparing side to side. No characteristic pattern of abnormalities on electromyography is noted in the cumulative stress cases. Patients with no motor symptoms, regardless of etiology, are more apt to have sensory distal latency prolongation, whereas those with motor symptoms have motor amplitude and needle examination abnormalities. CONCLUSIONS: Traumatic ulnar neuropathy at or distal to the wrist is characterized by motor symptoms and sensory and motor axonal loss by electromyography, whereas cumulative stress cases have sensory symptoms and electromyographic findings that are highly variable and noncharacteristic. Patients with no motor symptoms are more apt to show sensory distal latency abnormalities on electromyography, whereas those with motor symptoms show motor-evoked amplitude and needle electromyography abnormalities. PMID- 17398254 TI - Improving hand function in stroke survivors: a pilot study of contralaterally controlled functional electric stimulation in chronic hemiplegia. AB - OBJECTIVE: To assess the feasibility of a new stroke rehabilitation therapy for the hemiparetic hand. DESIGN: Case series. Pre- and postintervention assessment with 1- and 3-month follow-ups. SETTING: Clinical research laboratory of a large public hospital. PARTICIPANTS: Three subjects with chronic (>6mo postcerebrovascular accident) upper-extremity hemiplegia. INTERVENTION: Subjects used an electric stimulator to cause the paretic hand extensor muscles to contract and thereby open the hand. Subjects controlled the intensity of the stimulation, and thus the degree of hand opening, by volitionally opening the unimpaired contralateral hand, which was detected by an instrumented glove. For 6 weeks, subjects used the stimulator to perform active repetitive hand-opening exercises 2 hours daily at home and functional tasks 1.5 hours twice a week in the laboratory. MAIN OUTCOME MEASURES: Maximum voluntary finger extension, maximum voluntary isometric finger-extension moment, finger-movement control, and box and block test (BBT) score at pre- and posttreatment and at 1 month and 3 months posttreatment. RESULTS: Maximum voluntary finger extension increased from baseline to end of treatment and from the end of treatment to 1-month follow-up in 2 subjects. Maximum voluntary isometric finger-extension moment, finger movement control, and BBT score increased from baseline to the end of treatment and from the end of treatment to 1-month follow-up in all 3 subjects. The improvements generally declined at 3 months. CONCLUSIONS: The results suggest a positive effect on motor impairment, meriting further investigation of the intervention. PMID- 17398255 TI - Sequence of recovery during the course of emergence from the minimally conscious state. AB - OBJECTIVE: To document and examine recovery profiles of persons during the course of emergence from a minimally conscious state (MCS) after traumatic brain injury (TBI). DESIGN: Case series. SETTING: Participants in an inpatient brain injury rehabilitation program. PARTICIPANTS: Nine people with TBI who emerged from MCS. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: The Western Neuro Sensory Stimulation Profile. RESULTS: In all cases, stereotyped (nonreflexive) movement was the first behavior to resolve, yet by itself it was never sufficient to indicate emergence from the MCS. Two of the 9 patients showed consistent object manipulation before all 3 elements of functional communication. Conversely, in 2 patients, all 3 components of functional communication emerged before object manipulation. In the remaining 5 patients, object manipulation occurred along with components of functional communication. CONCLUSIONS: During the course of data collection, issues were raised about differentiation between the MCS and posttraumatic amnesia. The data are also discussed within the context of the work of Giacino et al and the development of their scale to measure duration of the MCS. These data support the theory that functional communication and object manipulation are the 2 key indicators of emergence from MCS. PMID- 17398256 TI - Priorities for stroke rehabilitation and research: results of a 2003 Canadian Stroke Network consensus conference. AB - The Canadian Stroke Network Consensus Conference panel met in 2003 in Toronto, Canada, to address areas of stroke rehabilitation that require additional research as well as increased efforts to knowledge translation. The results of an extensive literature review, of a study of factors related to poststroke quality of life, and a survey of clinicians were presented to the panel. From this review, the panel compiled a consensus list of 5 priority areas in stroke rehabilitation research that warrant further investigation. The priorities are: (1) multimodal programs for reintegration into the community; (2) rehabilitation of patients with severe strokes; (3) the ideal timing and intensity of aphasia therapy; (4) cognitive rehabilitation; and (5) and the timing and intensity of rehabilitation after mild-to-moderate stroke. The panel recommended that agencies that fund research create special competitions to support large, multicenter randomized controlled studies to investigate these areas of research. In addition, the panel identified 3 priority areas for knowledge translation where research was convincing: (1) lower-extremity (leg) interventions; (2) upper extremity (arm) interventions; and (3) detection of clients who are at risk of complications, specifically depression, dysphagia, or cognitive impairment, as well as those at risk for falls and pressure ulcers. PMID- 17398257 TI - Response shift theory: important implications for measuring quality of life in people with disability. AB - Measurement of health-related quality of life (HRQOL) in people with disability can be problematic. Ambiguous or paradoxical findings can occur because of differences among people or changes within people regarding internal standards, values, or conceptualization of HRQOL. These "response shifts" can affect standard psychometric indices, such as reliability and validity. Attending to appraisal processes and response shift theory can inform development of HRQOL measures for people with disability that do not confound function and health and that consider important causal indicators such as environment. By design, most HRQOL measures equate function with health, necessarily leading to a lower measured HRQOL in people with functional impairments regardless of their level of self-perceived health. In this article, we present theoretical and conceptual distinctions building on response shift theory and other current developments in HRQOL research. We then submit a set of suggested directions for future measurement development in populations with disabilities that consider these distinctions and extend their use in future measurement developments. PMID- 17398258 TI - Expert opinion and controversies in sports and musculoskeletal medicine: the diagnosis and treatment of spondylolysis in adolescent athletes. AB - Although spondylolysis is relatively common in adolescent athletes, there are substantial disagreements in the literature concerning the best methods for diagnosing and treating the condition. Controversy particularly arises regarding the optimal use of available imaging modalities in the diagnosis of athletes with suspected pars defects and the extent of activity restriction or brace use required for appropriate treatment. Because there have been no controlled trials on the treatment of spondylolysis and only a very limited number of studies addressing potential imaging strategies, it is difficult to develop true evidence based guidelines for this condition. Given the current state of the literature, it is our impression that nuclear imaging with single photon emission computed tomography followed by computed tomography, with a limited role for plain radiography, remains the standard for appropriately diagnosing a symptomatic pars lesion. Treatment hinges on activity restriction for an amount of time adequate to allow for symptom resolution and, when possible, potential bony healing followed by a progressive sport-specific rehabilitation program. The biomechanic effects of brace use in this population are not well understood, but there may be some detrimental effects to the use of a brace and there currently is no evidence that the routine use of a rigid brace results in any significant improvement in radiographic or functional outcome. PMID- 17398260 TI - Botulinum toxin type A and other factors. PMID- 17398261 TI - Parental evaluation and botulinum toxin type A. PMID- 17398263 TI - Ischaemic and non-ischaemic cardiomyopathies--cardiac MRI appearances with delayed enhancement. AB - Over the past few years, cardiovascular magnetic resonance (CMR) imaging has rapidly developed and is now a robust clinical tool capable of providing high resolution images of the heart in any desired plane. Delayed contrast-enhanced CMR (DE-CMR) can be used for non-invasive tissue characterization, with differing patterns of hyperenhancement displayed by ischaemic and non-ischaemic cardiomyopathies. This review explains the theory behind delayed hyperenhancement, and demonstrates the potential of DE-CMR in the diagnosis of a wide range of different cardiac disease states. PMID- 17398264 TI - Cranial CT of the unconscious adult patient. AB - Unconscious patients are frequently referred to radiology departments for computed tomography (CT) of the brain. The objectives of these examinations are to define the underlying cause of impaired consciousness and in some cases to determine the severity of associated brain injury. There is often little history available to guide the clinician or radiologist, particularly in patients referred from the casualty department. In this review, we present the typical CT appearances of adult patients presenting with loss of consciousness for CT examination. We focus on the most common abnormalities that are identified in everyday radiological practice, emphasize important diagnostic signs that may enable a confident diagnosis to be made and discuss when further imaging may be warranted. PMID- 17398265 TI - MRI of the rotator interval of the shoulder. AB - The rotator interval of the shoulder joint is located between the distal edges of the supraspinatus and subscapularis tendons and contains the insertions of the coracohumeral and superior glenohumeral ligaments. These structures form a complex pulley system that stabilizes the long head of the biceps tendon as it enters the bicipital groove of the humeral head. The rotator interval is the site of a variety of pathological processes including biceps tendon lesions, adhesive capsulitis and anterosuperior internal impingement. This article describes the anatomy, function and pathology of the rotator interval using magnetic resonance imaging (MRI). PMID- 17398266 TI - CT colonography: interpretative performance in a non-academic environment. AB - AIM: To investigate interpretative accuracy and reporting time for radiologists performing computed tomography (CT) colonography in day-to-day non-academic clinical practice. MATERIALS AND METHODS: Thirteen radiologists from seven centres, who were reporting CT colonography in non-academic daily clinical practice, interpreted a dataset of 15 colonoscopically validated cases in a controlled environment. Ten cases had either a cancer or polyp >10mm; one case had a medium polyp and four were normal. Correct case categorization and interpretation times were compared using analysis of variance to aggregated results obtained from both experienced observers and observers recently trained using 50 cases, working in an academic environment. The effect of experience was determined using Spearman's rank correlation. RESULTS: Individual accuracy was highly variable, range 53% (95% CI 27-79%) to 93% (95% CI 68-100%). Mean accuracy overall was significantly inferior to experienced radiologists (mean 75 versus 88%, p=0.04) but not significantly different from recently trained radiologists (p=0.48). Interpretation time was not significantly different to experienced readers (mean 12.4 min versus 11.7, p=0.74), but shorter than recently trained radiologists (p=0.05). There was a significant, positive, linear correlation between prior experience and accuracy (p<0.001) with no plateau. CONCLUSION: Accuracy for sub-specialist radiologists working in a non-academic environment is, on average, equivalent to radiologists trained using 50 cases. However, there is wide variability in individual performance, which generally falls short of the average performance suggested by meta-analysis of published data. Experience improves accuracy, but alone is insufficient to determine competence. PMID- 17398268 TI - CT of pelvic extraperitoneal spaces: an anatomical study in cadavers. AB - AIM: To evaluate the pelvic extraperitoneal compartments and communications with abdominal retroperitoneal spaces. MATERIAL AND METHODS: Helical computed tomography (CT) was used to image the abdomen and pelvis after injection of 800 ml of dilute (1 in 25) contrast material into prevesical, perivesical and perirectal spaces in eight embalmed cadavers. Axial images and multiplanar reconstructions were reviewed to determine flow pathways. RESULTS: The prevesical space was injected in four cadavers, the perivesical space in two and the perirectal in two. After the four prevesical space injections, communication was seen with the perivesical (four of four), perirectal (one of four) and abdominal extraperitoneal spaces (posterior pararenal space in all, anterior pararenal space in two of four, and perirenal space in three of four). After the two perivesical injections, communication was seen with the prevesical (two of two), perirectal (two of two) and abdominal extraperitoneal spaces (posterior pararenal in two of two, anterior pararenal in two of two, and perirenal space in two of two). After the two perirectal space injections, communication was seen with the prevesical (two of two), perivesical (one of two) and abdominal extraperitoneal spaces (posterior pararenal in two of two, anterior pararenal in two of two, and perirenal space in one of two). CONCLUSION: The extraperitoneal spaces of the pelvis comprise three communicating compartments: the prevesical space, the perivesical space, and the perirectal space. The perirectal space, previously thought to be separate, communicates with the perivesical and the prevesical spaces. Intercommunication occurs both between the pelvic extraperitoneal spaces and with abdominal retroperitoneal spaces. PMID- 17398269 TI - The influence of HRT on technical recall in the UK Breast Screening Programme: are pain, compression force, and compressed breast thickness contributing factors? AB - AIM: To investigate recall for technical reasons within the UK Breast Screening Programme, and to determine whether differences exist in those women using hormone replacement therapy (HRT), considering potential associations with reported pain, compression force used and compressed breast thickness (CBT) obtained. MATERIALS AND METHOD: A prospective cohort study of 2765 women attending for incident round breast screening appointments who were either HRT users, with a minimum of 1 year duration (n=1077), or had never used HRT (n=1688). Data were collected using technical recall records, a radiographer data collection sheet, and a self-administered participant questionnaire. RESULTS: Sixty-eight (2.5%) participants were recalled for technical reasons of whom 28 (2.6%) were HRT and 40 (2.4%) non-HRT users. This difference was not statistically significant (p=0.80). Significant differences were found for CBT between those HRT users who were and were not recalled for technical reasons (p<0.01) and for the similar categories of non-HRT users (p=0.03). No significant differences were found for force between those HRT users who were recalled or not (p=0.73) and for the similar categories of non-HRT users (p=0.07). Similarly no significant differences were found for pain between those HRT users who were recalled or not (p=0.75) and for the similar categories of non-HRT users (p=0.73). CONCLUSION: CBT was the only variable to have both a statistically and a clinically significant relationship with technical recall. PMID- 17398270 TI - Early stroke detection and extent: impact of experience and the role of computed tomography angiography source images. AB - AIM: To test the performance of computed tomography angiography "source images" (CTA-SI) versus unenhanced CT (NCCT) for stroke detection and extent using the Alberta Stroke Programme Early CT Score (ASPECTS), and examine the effect of experience and clinical history. MATERIALS AND METHODS: Studies of 23 consecutive patients presenting within 4.5h were analysed by three reviewers of varying experience. Each reviewer, blinded to clinical information reviewed a random order of NCCT and CTA-SI and documented side of infarct and the ASPECTS. The readings were repeated for CTA-SI with and without clinical information. Performance measures and observer agreement were calculated. Applying an ASPECTS threshold of20 years, due to its high activity against both adult and juvenile flukes. More recently, it has been used successfully to treat human cases of fascioliasis. Resistance to TCBZ first appeared in the field in Australia in the mid-1990s. Since then, resistance has been reported from a number of countries throughout Europe: Ireland, Scotland, Wales, Spain and The Netherlands. The heavy reliance on a single drug puts treatment strategies for fascioliasis at risk. Should resistance develop further, the prospect is an alarming one. This review will present an overview of progress in understanding the mechanism of resistance to TCBZ, examining possible changes in the target molecule, in drug influx/efflux mechanisms and in the metabolism of TCBZ by the fluke. The review will also consider ways to deal with resistance, covering drug oriented options such as: the use of alternative drugs, drug combinations and the search for new compounds. PMID- 17398284 TI - Primary care in the USA. PMID- 17398282 TI - Binding of the Alzheimer amyloid beta-peptide to neuronal cell membranes by fluorescence correlation spectroscopy. AB - The deposition of the Alzheimer amyloid beta-peptide (Abeta) fibrils in brain is a key step in Alzheimer's disease. The aggregated Abeta is found to be toxic to neurons since cells die when the aggregated Abeta is added to the cell culture medium. However, target of action of Abeta to cells is unknown. We have applied the fluorescence correlation spectroscopy (FCS) technique to study the existence of a receptor or target molecule for the Alzheimer amyloid beta-peptide (Abeta) in cultured human cerebral cortical neurons. FCS measurement of the fluorophore rhodamine-labeled Abeta (Rh-Abeta) shows diffusion times: 0.1 ms, 1.1 ms and 5.9 ms. Thus, 0.1 ms corresponds to the unbound Rh-Abeta, and 1.1 ms and 5.9 ms correspond to slowly diffusing complexes of Rh-Abeta bound to a kind of receptor or target molecule for Abeta. Addition of excess non-labeled Abeta is accompanied by a competitive displacement, showing that the Abeta binding is specific. Full saturation of the Abeta binding is obtained at nanomolar concentrations, indicating that the Abeta binding is of high affinity. The notion that using FCS we have found a kind of receptor or target molecule for Abeta makes an important point that Abeta kills cells possibly by affecting cell membranes via a receptor or target molecule. This study is of highly significance since it suggests that Abeta possibly affects neuronal cell membranes of Alzheimer patients via a receptor or target molecule. PMID- 17398285 TI - New concerns about oseltamivir. PMID- 17398286 TI - Food crisis in Cambodia. PMID- 17398287 TI - Adolescent health: an opportunity not to be missed. PMID- 17398288 TI - Us and them: worldwide health issues for adolescents. PMID- 17398289 TI - Steroids to prevent postextubation laryngeal oedema. PMID- 17398290 TI - JELIS, fish oil, and cardiac events. PMID- 17398291 TI - Occipital nerve stimulation for intractable cluster headache. PMID- 17398292 TI - Exclusive breastfeeding and HIV. PMID- 17398294 TI - A code of ethics for scientists. PMID- 17398295 TI - Elizabeth Mapella: prioritising adolescent health in Tanzania. PMID- 17398296 TI - Umbilical cord blood banking and the RCOG. PMID- 17398297 TI - Statins for primary prevention of coronary artery disease. PMID- 17398298 TI - Statins for primary prevention of coronary artery disease. PMID- 17398299 TI - Statins for primary prevention of coronary artery disease. PMID- 17398301 TI - Treating stroke in the 21st century. PMID- 17398302 TI - Treatment of narcolepsy with cataplexy. PMID- 17398303 TI - Treating stroke in the 21st century. PMID- 17398304 TI - Low birthweight and subsequent obesity in Japan. PMID- 17398305 TI - Treatment of narcolepsy with cataplexy. PMID- 17398306 TI - Potential applicability of ABCD score in triaging TIA patients. PMID- 17398307 TI - 12-h pretreatment with methylprednisolone versus placebo for prevention of postextubation laryngeal oedema: a randomised double-blind trial. AB - BACKGROUND: The efficacy of corticosteroids in reducing the incidence of postextubation laryngeal oedema is controversial. We aimed to test our hypothesis that methylprednisolone started 12 h before a planned extubation could prevent postextubation laryngeal oedema. METHODS: We did a placebo-controlled, double blind multicentre trial in 761 adults in intensive-care units. Patients who were ventilated for more than 36 h and underwent a planned extubation received intravenous 20 mg methylprednisolone (n=380) or placebo (381) 12 h before extubation and every 4 h until tube removal. The primary endpoint was occurrence of laryngeal oedema within 24 h of extubation. Laryngeal oedema was clinically diagnosed and deemed serious if tracheal reintubation was needed. Analyses were done on a per protocol and intention-to-treat basis. This trial is registered at ClinicalTrials.gov, number NCT00199576. FINDINGS: 63 patients could not be assessed, mainly because of self-extubation (n=16) or cancelled extubation (44) between randomisation and planned extubation. 698 patients were analysed (343 in placebo group, 355 in methylprednisolone group). Methylprednisolone significantly reduced the incidence of postextubation laryngeal oedema (11 of 355, 3%vs 76 of 343, 22%, p<0.0001), the global incidence of reintubations (13 of 355, 4%vs 26 of 343, 8%, p=0.02), and the proportion of reintubations secondary to laryngeal oedema (one of 13, 8 %vs 14 of 26, 54%, p=0.005). One patient in each group died after extubation, and atelectasia occurred in one patient given methylprednisolone. INTERPRETATION: Methylprednisolone started 12 h before a planned extubation substantially reduced the incidence of postextubation laryngeal oedema and reintubation. Such pretreatment should be considered in adult patients before a planned extubation that follows a tracheal intubation of more than 36 h. PMID- 17398308 TI - Effects of eicosapentaenoic acid on major coronary events in hypercholesterolaemic patients (JELIS): a randomised open-label, blinded endpoint analysis. AB - BACKGROUND: Epidemiological and clinical evidence suggests that an increased intake of long-chain n-3 fatty acids protects against mortality from coronary artery disease. We aimed to test the hypothesis that long-term use of eicosapentaenoic acid (EPA) is effective for prevention of major coronary events in hypercholesterolaemic patients in Japan who consume a large amount of fish. METHODS: 18 645 patients with a total cholesterol of 6.5 mmol/L or greater were recruited from local physicians throughout Japan between 1996 and 1999. Patients were randomly assigned to receive either 1800 mg of EPA daily with statin (EPA group; n=9326) or statin only (controls; n=9319) with a 5-year follow-up. The primary endpoint was any major coronary event, including sudden cardiac death, fatal and non-fatal myocardial infarction, and other non-fatal events including unstable angina pectoris, angioplasty, stenting, or coronary artery bypass grafting. Analysis was by intention-to-treat. The study was registered at ClinicalTrials.gov, number NCT00231738. FINDINGS: At mean follow-up of 4.6 years, we detected the primary endpoint in 262 (2.8%) patients in the EPA group and 324 (3.5%) in controls-a 19% relative reduction in major coronary events (p=0.011). Post-treatment LDL cholesterol concentrations decreased 25%, from 4.7 mmol/L in both groups. Serum LDL cholesterol was not a significant factor in a reduction of risk for major coronary events. Unstable angina and non-fatal coronary events were also significantly reduced in the EPA group. Sudden cardiac death and coronary death did not differ between groups. In patients with a history of coronary artery disease who were given EPA treatment, major coronary events were reduced by 19% (secondary prevention subgroup: 158 [8.7%] in the EPA group vs 197 [10.7%] in the control group; p=0.048). In patients with no history of coronary artery disease, EPA treatment reduced major coronary events by 18%, but this finding was not significant (104 [1.4%] in the EPA group vs 127 [1.7%] in the control group; p=0.132). INTERPRETATION: EPA is a promising treatment for prevention of major coronary events, and especially non-fatal coronary events, in Japanese hypercholesterolaemic patients. PMID- 17398309 TI - Treatment of medically intractable cluster headache by occipital nerve stimulation: long-term follow-up of eight patients. AB - BACKGROUND: Cluster headache is a form of primary headache that features repeated attacks of excruciatingly severe headache usually occurring several times a day. Patients with chronic cluster headache have unremitting illness that necessitates daily preventive medical treatment for years. When medically intractable, the condition has previously been treatable only with cranially invasive or neurally destructive methods. METHODS: Eight patients with medically intractable chronic cluster headache were implanted in the suboccipital region with electrodes for occipital nerve stimulation. Other than the first patient, who was initially stimulated unilaterally before being stimulated bilaterally, all patients were stimulated bilaterally during treatment. FINDINGS: At a median follow-up of 20 months (range 6-27 months for bilateral stimulation), six of eight patients reported responses that were sufficiently meaningful for them to recommend the treatment to similarly affected patients with chronic cluster headache. Two patients noticed a substantial improvement (90% and 95%) in their attacks; three patients noticed a moderate improvement (40%, 60%, and 20-80%) and one reported mild improvement (25%). Improvements occurred in both frequency and severity of attacks. These changes took place over weeks or months, although attacks returned in days when the device malfunctioned (eg, with battery depletion). Adverse events of concern were lead migrations in one patient and battery depletion requiring replacement in four. INTERPRETATION: Occipital nerve stimulation in cluster headache seems to offer a safe, effective treatment option that could begin a new era of neurostimulation therapy for primary headache syndromes. PMID- 17398310 TI - Mother-to-child transmission of HIV-1 infection during exclusive breastfeeding in the first 6 months of life: an intervention cohort study. AB - BACKGROUND: Exclusive breastfeeding, though better than other forms of infant feeding and associated with improved child survival, is uncommon. We assessed the HIV-1 transmission risks and survival associated with exclusive breastfeeding and other types of infant feeding. METHODS: 2722 HIV-infected and uninfected pregnant women attending antenatal clinics in KwaZulu Natal, South Africa (seven rural, one semiurban, and one urban), were enrolled into a non-randomised intervention cohort study. Infant feeding data were obtained every week from mothers, and blood samples from infants were taken monthly at clinics to establish HIV infection status. Kaplan-Meier analyses conditional on exclusive breastfeeding were used to estimate transmission risks at 6 weeks and 22 weeks of age, and Cox's proportional hazard was used to quantify associations with maternal and infant factors. FINDINGS: 1132 of 1372 (83%) infants born to HIV-infected mothers initiated exclusive breastfeeding from birth. Of 1276 infants with complete feeding data, median duration of cumulative exclusive breastfeeding was 159 days (first quartile [Q1] to third quartile [Q3], 122-174 days). 14.1% (95% CI 12.0 16.4) of exclusively breastfed infants were infected with HIV-1 by age 6 weeks and 19.5% (17.0-22.4) by 6 months; risk was significantly associated with maternal CD4-cell counts below 200 cells per muL (adjusted hazard ratio [HR] 3.79; 2.35-6.12) and birthweight less than 2500 g (1.81, 1.07-3.06). Kaplan-Meier estimated risk of acquisition of infection at 6 months of age was 4.04% (2.29 5.76). Breastfed infants who also received solids were significantly more likely to acquire infection than were exclusively breastfed children (HR 10.87, 1.51 78.00, p=0.018), as were infants who at 12 weeks received both breastmilk and formula milk (1.82, 0.98-3.36, p=0.057). Cumulative 3-month mortality in exclusively breastfed infants was 6.1% (4.74-7.92) versus 15.1% (7.63-28.73) in infants given replacement feeds (HR 2.06, 1.00-4.27, p=0.051). INTERPRETATION: The association between mixed breastfeeding and increased HIV transmission risk, together with evidence that exclusive breastfeeding can be successfully supported in HIV-infected women, warrant revision of the present UNICEF, WHO, and UNAIDS infant feeding guidelines. PMID- 17398311 TI - Jet lag: trends and coping strategies. AB - The number of travellers undertaking long-distance flights has continued to increase. Such flights are associated with travel fatigue and jet lag, the symptoms of which are considered here, along with their similarities, differences, and causes. Difficulties with jet lag because of sleep loss and decreased performance are emphasised. Since jet lag is caused mainly by inappropriate timing of the body clock in the new time zone, the pertinent properties of the body clock are outlined, with a description of how the body clock can be adjusted. The methods, both pharmacological and behavioural, that have been used to alleviate the negative results of time-zone transitions, are reviewed. The results form the rationale for advice to travellers flying in different directions and crossing several time zones. Finally, there is an account of the main problems that remain unresolved. PMID- 17398312 TI - Pubertal transitions in health. AB - Puberty is accompanied by physical, psychological, and emotional changes adapted to ensure reproductive and parenting success. Human puberty stands out in the animal world for its association with brain maturation and physical growth. Its effects on health and wellbeing are profound and paradoxical. On the one hand, physical maturation propels an individual into adolescence with peaks in strength, speed, and fitness. Clinicians have viewed puberty as a point of maturing out of childhood-onset conditions. However, puberty's relevance for health has shifted with a modern rise in psychosocial disorders of young people. It marks a transition in risks for depression and other mental disorders, psychosomatic syndromes, substance misuse, and antisocial behaviours. Recent secular trends in these psychosocial disorders coincide with a growing mismatch between biological and social maturation, and the emergence of more dominant youth cultures. PMID- 17398313 TI - Endogenous lipoid pneumonia associated with primary sclerosing cholangitis. PMID- 17398315 TI - Systemic hypertension. AB - Hypertension is a growing public health problem worldwide. Only 37% of American hypertensives currently have their blood pressures controlled. Hypertension is traditionally diagnosed in the medical office, but both home and ambulatory blood pressure monitoring can help. Lifestyle modifications are recommended for everyone who has higher than "normal" blood pressure (<120/80 mm Hg). Voluminous clinical trial data support beginning drug therapy with low-dose chlorthalidone, unless the patient has a specific indication for a different drug. Additional drugs (typically in the sequence, angiotensin converting-enzyme inhibitor or angiotensin receptor blocker, calcium antagonist, beta-blocker, alpha-blocker, aldosterone antagonist, direct vasodilator, and centrally acting alpha(2) agonist) can be added to achieve the blood pressure goal (usually <140/90 mm Hg, but <130/80 mm Hg for diabetics and those with chronic kidney disease). Special circumstances exist for treatment of hypertension in pregnancy, in childhood, in the elderly, and in both extremes of blood pressure (pre-hypertension or hypertensive emergencies). PMID- 17398316 TI - Keratoplasty for endothelial dysfunction. PMID- 17398317 TI - A prospective study of endothelial cell loss during the 2 years after deep lamellar endothelial keratoplasty. AB - PURPOSE: To report the endothelial survival over a 2-year period after 2 techniques of deep lamellar endothelial keratoplasty (DLEK) in the treatment of endothelial dysfunction. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: One hundred eyes of 88 patients with corneal edema. METHODS: One hundred consecutive eyes with endothelial failure were entered into a prospective study of endothelial keratoplasty, and the donor central endothelial cell density (ECD) was recorded postoperatively at 6 months (n = 98), 12 months (n = 96), and 24 months (n = 85) and then compared with the preoperative eye bank measurements. The subsets of eyes with large-incision DLEK (n = 36) and small-incision DLEK (n = 62) were also evaluated and compared. MAIN OUTCOME MEASURES: Preoperative and postoperative central ECDs were prospectively evaluated and the cell loss calculated for each postoperative time point. RESULTS: The average (and standard deviation) ECD at 6 months was 2140+/-426 cells/mm(2), representing a mean cell loss from preoperative donor cell measurements of 25+/-15%. At 12 months, ECD was 2090+/-448 cells/mm2 (26+/-16% cell loss), and at 24 months, it was 1794+/-588 cells/mm2 (37+/-27% cell loss). The additional cell loss from 1 to 2 years was significant (P<0.001). In the subset of large-incision DLEK eyes (n = 36), the cell loss from preoperatively to 6 months was 23%; 12 months, 22%; and 24 months, 27%. In the subset of small incision DLEK eyes (n = 62), the cell loss from preoperatively to 6 months was 25%; 12 months, 28%; and 24 months, 43%. The cell loss from small-incision DLEK surgery was significantly greater than that from large-incision DLEK surgery at the 12-month (P = 0.013) and 24-month (P<0.001) postoperative measurements. CONCLUSIONS: Although the initial cell loss from DLEK surgery is minimally changed from 6 to 12 months postoperatively, there is an acceleration of cell loss from 1 year to 2 years postoperatively. The small-incision DLEK technique, which involves folding of the donor tissue, results in a significantly higher endothelial cell loss at 1 and 2 years than that found after large-incision DLEK surgery, wherein the tissue is not folded. PMID- 17398318 TI - Accounting for restart rates in evaluating persistence with ocular hypotensives. AB - PURPOSE: Persistence with ocular hypotensive medication is important as a long term outcome, and rates of persistence typically are low. This study assessed restart rates for 3 prostaglandin analogs by determining the percentage of patients who discontinued and then restarted therapy. DESIGN: Retrospective cohort study of pharmacy claims submitted to a large national administrative claims database. PARTICIPANTS: In all, 4356 patients initiating prostaglandin therapy were identified. METHODS: Claims for 3 prostaglandin analogs (bimatoprost, latanoprost, travoprost [index prostaglandin]) submitted between 2001 and 2002 were analyzed. Patients were excluded if they did not have coverage in the plan for the preceding 180 days or had been prescribed any ocular prostaglandin in the prior 180 days. MAIN OUTCOME MEASURES: Persistence was defined as neither discontinuing nor changing the index prostaglandin. The number of current users of the index prostaglandin at day 180 was the sum of patients who persisted with the index prostaglandin plus patients who restarted the index prostaglandin after a discontinuation. RESULTS: Of the 4356 patients initiating prostaglandin therapy, 2503 (57%) were potential current users (were still plan members and had not switched ocular hypotensive therapies after 180 days). Just over half, (1356/2503 [54%]) were current users, including 879 (35%) who persisted with their index prostaglandin and 477 (19%) who restarted their index prostaglandin. Of patients discontinuing their index prostaglandin, more than half failed to restart any topical therapy (827/1624 [51%]). CONCLUSIONS: Previous studies showing low persistence rates for glaucoma therapy failed to evaluate restarts. Restart analyses are crucial for assessing long-term treatment of chronic diseases such as glaucoma. In general, persistence remains a challenge, and our findings demonstrate the importance of clinicians' identifying patients who are not persistent and encouraging them either to restart or to initiate treatment with an alternative therapy. PMID- 17398319 TI - A new pattern electroretinogram paradigm evaluated in terms of user friendliness and agreement with perimetry. AB - PURPOSE: To assess ease of use of the pattern electroretinogram optimized for glaucoma screening (PERGLA) paradigm by a novice operator; to study test-retest variability of the PERGLA parameters; and to compare results from the PERGLA to those from perimetry. DESIGN: Cohort study. PARTICIPANTS: Twelve healthy control subjects and 16 patients with moderate to advanced glaucoma in at least 1 eye. METHODS: Pattern electroretinograms were recorded simultaneously from both eyes using a commercially available testing station. Each participant underwent PERGLA procedures in 2 sessions. One eye of each subject was tested on contrast sensitivity perimetry (CSP) in which a 0.4 cycles/degree Gabor patch served as a stimulus. Central visual fields results from conventional automated perimetry (CAP) were obtained from patients' records. Bland-Altman analysis was performed on PERGLA results to assess normal test-retest variability. Differences from mean normal (in decibels [dB]) were compared for PERGLA versus CSP and CAP. MAIN OUTCOME MEASURES: Pattern electroretinogram amplitude, noise, phase, and test retest variability (coefficient of variation); contrast sensitivity from CSP; perimetric sensitivity from CAP; and differences from mean normal for PERGLA, CSP, and CAP. RESULTS: The mean log amplitude (0.08+/-0.12 log muV) and the mean phase (1.92+/-0.07 pi rad) for the control group were consistent with published PERGLA norms, as was test-retest variability for both amplitude (coefficient of variation [CV] = 8.2+/-7.0%) and phase (CV = 1.1+/-0.9%). The mean signal-to noise ratio (8.7+/-4.5) was lower than published norms. The test-retest variability increased as PERGLA log amplitude decreased (R2>0.12, P<0.05). On average, differences from mean normal were similar for PERGLA versus CSP and for PERGLA versus CAP (mean differences<0.5 dB) with 95% confidence intervals near +/ 4 dB in both comparisons. CONCLUSIONS: A novice operator successfully replicated published PERGLA norms in a young control group for amplitude, phase, and repeatability. Higher test-retest variability was found in eyes with smaller signals. On average, PERGLA results were in reasonable agreement with those from perimetry, although there existed large individual differences which may limit the usage of PERGLA in screening or in following progression of glaucoma. PMID- 17398320 TI - Randomized controlled trial of combined 5-Fluorouracil and low-molecular-weight heparin in the management of unselected rhegmatogenous retinal detachments undergoing primary vitrectomy. AB - OBJECTIVE: To determine the efficacy of a combination of 5-fluorouracil (5FU) and low-molecular-weight heparin (LMWH) in the treatment of unselected rhegmatogenous retinal detachment (RRD) undergoing primary vitrectomy. DESIGN: Double-masked, prospective, randomized, placebo-controlled clinical trial. PARTICIPANTS: Six hundred forty-one patients presenting with primary RRD were recruited from 2 specialized vitreoretinal units-Moorfields Eye Hospital, London (n = 553) and St. Pauls Eye Unit, Liverpool (n = 88). INTERVENTION: All patients underwent primary vitrectomy and gas endotamponade. Adjuvant therapy in the treatment group consisted of 5 IU/ml LMWH and 200 mug/ml 5FU added to the perioperative infusion fluid. MAIN OUTCOME MEASURES: The primary outcome measure was retinal reattachment after primary vitrectomy without any reoperations at 6 months. Secondary outcome measures recorded at 6 months were the occurrence and grade of proliferative vitreoretinopathy (PVR), best-corrected visual acuity in logarithm of the minimum angle of resolution, intraocular pressure (mmHg), corneal clarity, and complications. RESULTS: The overall primary success rate was 84.4%; in the treatment group, the primary success rate was 82.3% compared with 86.8% in the placebo group (P = 0.12). At 6 months, the final complete anatomical reattachment rate was 97.9% in both treatment and placebo groups. The number of patients who failed due to the development of PVR was not statistically significant, 23 in the treatment group (7.0%) and 14 in the placebo group (4.9%) (P = 0.309). There was no significant difference in the mean visual acuity at 6 months in the placebo group (0.48) versus the treatment group (0.53; P = 0.072). The visual acuity at 6 months of patients presenting with a macula-sparing retinal detachment was significantly worse in the treatment group (P = 0.0091). There was no significant difference between the 2 groups in patients who presented with a macula involving retinal detachment (P = 0.896). CONCLUSIONS: Primary vitrectomy has a high anatomic and visual success rate for RRD. Adjuvant therapy with 5FU and LMWH does not improve the anatomic or visual success rate of unselected primary retinal detachments undergoing vitrectomy. After adjuvant therapy, a worse visual outcome was observed in patients presenting with macula-sparing retinal detachments. A combination of 5FU and LMWH should not be used routinely for primary RRD surgery. PMID- 17398321 TI - Association of complement factor H Y402H gene polymorphism with different subtypes of exudative age-related macular degeneration. AB - OBJECTIVE: Exudative age-related macular degeneration (AMD) is a common cause for a severe central visual loss. The complement system has been implicated in the pathogenesis of drusen. Recently, a complement factor H (CFH) polymorphism, which is characterized by a tyrosine (Y)-to-histidine (H) exchange at position 402 of the CFH gene, has been suggested as a major risk factor for AMD in a North American population. The aim of the present study was to investigate a hypothesized association between the CFH Y402H polymorphism and the presence of exudative AMD in a Central European population of Caucasoid descent as well as to determine the genotype distribution among different types of exudative AMD. DESIGN: Retrospective case-control study. PARTICIPANTS: The study cohort consisted of 179 patients with exudative AMD and 163 controls. METHODS: Determination of genotypes was carried out by allele-specific digestion of polymerase chain reaction products. MAIN OUTCOME MEASURES: Genotypes of CFH Y402H polymorphism. RESULTS: The prevalence of the CFH 402HH genotype was significantly higher in patients with exudative AMD than among controls (35.2% vs. 8.6%; P<0.001). Homozygosity for the CFH Y402H polymorphism was associated with an odds ratio of 5.78 (95% confidence interval, 3.09-10.83) for exudative AMD. Subgroup analysis revealed that the CFH 402HH genotype was significantly more prevalent in eyes with predominantly classic with no occult choroidal neovascularization (CNV) than in those with either retinal angiomatous proliferation, occult with no classic CNV, or predominantly classic with occult CNV. CONCLUSION: Our data suggest that the CFH Y402H polymorphism is a major risk factor for exudative AMD in a Central European population. PMID- 17398322 TI - Primary intravitreal bevacizumab (Avastin) for diabetic macular edema: results from the Pan-American Collaborative Retina Study Group at 6-month follow-up. AB - PURPOSE: To report the 6-month anatomic and best-corrected visual acuity (BCVA) response after primary intravitreal bevacizumab (Avastin) in patients with diabetic macular edema (DME). DESIGN: Interventional retrospective multicenter study at 6 centers from 6 countries of patients with DME. PARTICIPANTS: We reviewed the clinical records of 88 consecutive patients (110 eyes) with DME. Seventy-eight eyes of 64 consecutive patients with a minimum follow-up of 6 months and mean age of 59.7+/-9.3 years were included in this analysis. INTERVENTION: Patients were treated with at least one intravitreal injection of 1.25 mg or 2.5 mg of bevacizumab and underwent Early Treatment Diabetic Retinopathy Study (ETDRS) BCVA testing, ophthalmoscopic examination, optical coherence tomography (OCT), and fluorescein angiography (FA) at baseline and follow-up visits. Repeated-measures analysis of variance was used to compare mean values. MAIN OUTCOME MEASURES: Changes in BCVA, OCT, and FA. RESULTS: Mean follow up was 6.31+/-0.81 months (range, 6-9). Sixteen (20.5%) eyes needed a second injection at a mean of 13.8 weeks (range, 4-28), and 6 eyes needed a third injection (7.7%) at a mean of 11.5 weeks (range, 5-20). The mean baseline BCVA was 0.87 (logarithm of the minimum angle of resolution), and the final mean BCVA was 0.6, a difference that was statistically significant (P<0.0001). Final BCVA analysis by subgroups demonstrated that 32 (41.1%) eyes remained stable, 43 (55.1%) improved > or =2 ETDRS lines of BCVA, and 3 (3.8%) decreased > or =2 ETDRS lines of BCVA. Mean central macular thickness at baseline by OCT was 387.0+/-182.8 mum and decreased to a mean of 275.7+/-108.3 at end of follow-up (P<0.0001). No ocular or systemic adverse events were observed. CONCLUSIONS: Primary intravitreal bevacizumab at doses of 1.25 to 2.5 mg seem to provide stability or improvement in VA, OCT, and FA in DME at 6 months. Follow-up is still short to make any specific treatment recommendations; however, the results appear promising. Evaluation in a multicenter randomized controlled clinical trial with longer follow-up is needed. PMID- 17398323 TI - Nonarteritic anterior ischemic optic neuropathy and tobacco smoking. AB - OBJECTIVE: To evaluate whether tobacco smoking may be a risk factor for development of nonarteritic anterior ischemic optic neuropathy (NA-AION). DESIGN: Cohort study. PARTICIPANTS: Six hundred twenty-four consecutive patients with NA AION who fulfilled our inclusion criteria. METHODS: All patients provided a detailed ocular, medical, and smoking history, and underwent a comprehensive ophthalmic evaluation. For data analysis, patients were divided into current, former, and never smokers. The prevalence for current smoking in the NA-AION patients was compared to the corresponding race-, gender-, age-, and period matched subgroup of the U.S. population. MAIN OUTCOME MEASURES: Association between NA-AION and tobacco smoking. RESULTS: Of the 624 patients, 369 (59.1%) were men, and mean (+/- standard deviation) age was 61.0+/-12.3 years. Of all the patients, 151 (24.2%) were current smokers, 160 (25.6%) former smokers, and 313 (50.2%) had never smoked. The prevalence of smoking in NA-AION patients was not significantly different from the prevalence in the period-matched U.S. population and the period matched Iowa population. In contrast, the prevalence of diabetes mellitus, ischemic heart disease, arterial hypertension, and cerebrovascular disease in NA-AION patients was significantly higher compared to the prevalence of these chronic conditions in the matched U.S. population (P<0.0001). There was no significant difference in initial visual acuity (P = 0.97) or the amount of initial visual field loss (P = 0.31) among nonsmokers, former smokers, and current smokers. Current smokers had NA-AION in the first eye at a significantly younger age (57.8+/-11.7 years) than former smokers (64.0+/-10.2 years; P<0.0001), and nonsmokers (60.4+/-13.5 years; P = 0.032). Comparison of the distribution of the time to develop NA-AION in the fellow eye showed no significant difference among the 3 groups (log-rank test P = 0.186). CONCLUSIONS: Our study showed no association between NA-AION and tobacco smoking. PMID- 17398324 TI - Impending recurrent pterygium. PMID- 17398325 TI - Impending recurrent pterygium. PMID- 17398327 TI - Resolution of scleromalacia. PMID- 17398329 TI - Toric IOL and postkeratoplasty astigmatism. PMID- 17398330 TI - Presbyopia after keratectomy. PMID- 17398332 TI - Idiopathic intracranial hypertension. PMID- 17398333 TI - 25-gauge vitrectomy. PMID- 17398334 TI - Virtual reality simulation. PMID- 17398335 TI - Resident surgical competency. PMID- 17398337 TI - Clinical decision support systems and computer-aided diagnosis in otology. AB - OBJECTIVES: We reviewed the progress of the implementation of expert diagnostic systems in the field of otology. STUDY DESIGN AND SETTING: We conducted a review of the literature at a research institute. RESULTS: The utilization of expert diagnostic systems in otology is very limited. Previous applications focused primarily upon the diagnosis of vertiginous disorders with the use of deterministic algorithms and, more recently, with adaptive algorithms such as neural networks. CONCLUSION: Expert systems provide greater diagnostic accuracy to physicians across a wide range of medical specialties. The success of such a system depends upon the strength of its reasoning algorithm, the validity of its knowledge base, and its ease of use. SIGNIFICANCE: There have been no attempts to develop an adaptive expert system for the full range of otological conditions. Such a tool may be of great use to physicians as a diagnostic aid and educational resource, particularly for those located in isolated sites. PMID- 17398338 TI - Post-tonsillectomy hemorrhage rates: are they technique-dependent? AB - OBJECTIVES: Recent publications have identified different rates of postoperative hemorrhage between "cold" tonsillectomy techniques and "hot" tonsillectomy techniques, generally identifying lower bleeding rates after cold techniques. Data from a prospective institutional review were analyzed to determine the relative risk factors for post-tonsillectomy hemorrhage among different techniques and by different age groups. MATERIALS AND METHODS: At the co-located John Hunter Hospital and John Hunter Children's Hospital, data were collected prospectively over a period of five years to allow a nonrandom comparison between a nondiathermy dissection technique for tonsillectomy with a monopolar diathermy technique in the management of nonmalignant disease of the tonsils, in children and adults, by determining complications in the first 28 days after surgery. RESULTS AND CONCLUSIONS: One thousand one hundred thirty-three consecutive cases were analyzed. The primary post-tonsillectomy hemorrhage rate was 0.2% for blunt dissection plus diathermy hemostasis and 0.3% for monopolar diathermy dissection plus hemostasis. Monopolar diathermy had a lower rate of secondary postoperative hemorrhage, requiring readmission (4.2% compared with 5.4% for blunt dissection plus diathermy hemostasis) and a lower rate for readmission for observation alone (2.1% compared with 4.2%) but had a higher risk of returning to surgery (1.6% compared with 1.04%) and a higher risk of blood transfusion (0.49% compared with 0.2%). These differences, however, did not reach statistical significance (Yates chi(2)), and neither did the relative risk between the two techniques. Two-way analysis of variance among secondary post-tonsillectomy hemorrhage complications by technique and by age groups shows a highly statistically significant difference by age group (analysis of variance, 3 df, F = 9.509, P < 0.001), much more so than technique. PMID- 17398339 TI - The wide nasal dorsum: evaluation and management. AB - OBJECTIVE: Rhinoplasty is arguably the most challenging of all facial aesthetic surgery. With contemporary aesthetic ideals favoring the more petite and narrow nose, successful management of the wide nasal dorsum is crucial in obtaining favorable outcomes. This paper aims to define the underlying anatomical variances that contribute to the wide nose, as well as providing a process by which one can identify and address these components appropriately. STUDY DESIGN AND SETTING: We conducted a retrospective study of 543 consecutive rhinoplasty patients presenting to a busy inner city cosmetic practice in Sydney, Australia. Of these patients, 53 were identified whose primary complaint was dissatisfaction with their wide nose. Analysis of the fundamental causes for these patients' wide nasal dorsum was carried out, and the techniques used to manage these problems were tabulated. Two illustrative case studies are included. RESULTS: Of the 53 patients, 29 (54.7%) could be managed by the simple technique of single lateral osteotomies in conjunction with medial osteotomies or hump removal. In the remainder, a variety of techniques were required to achieve the desired result. These included multiple lateral osteotomies (32.1%), wedge osteectomies (11.3%), nasal bone sculpturing (1.9%), reverse spreader technique (1.9%), cartilage sculpturing (3.8%), dorsal onlay grafting (22.6%), and deep fatty panniculus excision (35.8%). CONCLUSIONS: Management of the wide nasal dorsum should begin with a comprehensive analysis of the underlying causative anatomy. Although most cases may be managed satisfactorily with simple lateral osteotomies, a large percentage of patients presenting to the rhinoplasty surgeon will require adjunctive or alternate techniques to produce optimal results. PMID- 17398340 TI - Neck dissection for squamous cell carcinoma of the head and neck. AB - OBJECTIVES: To investigate the successes and failures of 172 patients receiving neck dissections for squamous cell carcinoma (SCC) managed through a multidisciplinary head and neck clinic and to observe factors in predicting failure (death with head and neck cancer or local regional recurrence) or local regional recurrence alone. STUDY DESIGN: A retrospective, 14-year surgical audit. METHODS: The information from patient medical records was correlated with that of a database. Multivariate analysis was performed with the use of a logistic regression model. RESULTS: The most common site for head and neck SCC was the oral cavity (42%), with 17% of patients having tumors of unknown primary site. The disease-free survival probability was 76.7% at five years. Patients who were older at neck dissection, were nonsmokers, or who did not have radiotherapy to the primary site were more likely to die with head and neck cancer or to have local regional recurrence (P < 0.1). Patients with more radical neck dissections did not have better survival or recurrence outcomes. Lymphatic invasion appears to be the only important factor in predicting local regional recurrence alone (P = 0.1), of which 67% occurred within the first postoperative year. CONCLUSIONS: Patients with smoking-related SCC are likely to have a less aggressive disease. Adjuvant radiotherapy plays an important role in the treatment of patients with head and neck SCC. Follow-up, especially within the first postoperative year, is essential in managing head and neck SCC. Head and neck surgeons can confidently continue their practice away from more radical neck dissections. PMID- 17398341 TI - Prediction of pharyngocutaneous fistulas after laryngectomy. AB - OBJECTIVE: To assess the predictive value of wound amylase as an indicator for pharyngocutaneous fistula development following laryngectomy for cancer. DESIGN AND SETTING: We conducted a prospective observational study at a tertiary referral center of 102 consecutive laryngeal or hypopharyngeal cancer patients undergoing laryngectomy with or without pharyngectomy. INTERVENTION: Data were collated on potential predictors of fistula formation compared with rate of development of clinical fistulas, all confirmed radiologically. MAIN OUTCOME MEASURES: Rate of fistula formation was determined for the following potential predictors: extent of resection (extended laryngectomy), postoperative wound (drain) amylase, previous radiotherapy, neck dissection, preoperative and postoperative hemoglobin and albumin levels, and postoperative transfusion. Sensitivity, specificity, and positive and negative predictive values of significant predictors were ascertained. RESULTS: The only significant predictors of fistula formation were extent of resection (extended laryngectomy) and drain amylase >4000 IU/L. If both factors are combined, the sensitivity, specificity, and positive and negative predictive values for fistula development are 83, 94, 63, and 98 percent, respectively. CONCLUSION: We advocate that patients be managed postoperatively according to the presence or absence of these given predictors to reduce occurrence of fistula formation in the high-risk group. PMID- 17398342 TI - The use of Monogen in the conservative management of chylous fistula. AB - BACKGROUND: Chylous fistula is an uncommon complication of lower neck dissection. If untreated, it can lead to severe electrolyte disturbance, fluid, and protein loss and ultimately threaten skin flaps and vital structures. Conservative management aims to decrease chyle volume by replacing long-chain triglycerides (LCTs) with medium-chain triglycerides in the diet or by total parenteral nutrition. METHODS: In 2001, Greenlane Head and Neck unit conducted a prospective 2-year study of all patients with chylous fistula. RESULTS: Eleven patients developed chylous fistula in the period between 2001 and 2003, out of 210 neck dissections. All cases were managed conservatively. The mean time to diagnosis was 1.5 days (1-4), time to closure was 8.1 days (4-26), and mean duration of treatment was 11.5 days (4-35). CONCLUSION: We would recommend conservative management of chylous fistulae with a low LCT diet using Monogen alone to be continued only for the duration of the fistula. PMID- 17398343 TI - Intraparotid lymphadenitis caused by Haemophilus aphrophilus. PMID- 17398344 TI - Post-tonsillectomy bleeding. PMID- 17398345 TI - Endoscopic treatment of fibrous dysplasia confined to the frontal sinus. PMID- 17398346 TI - Giant cell tumor of the mandible in a patient with Paget's disease. PMID- 17398347 TI - A case of renal cell carcinoma metastasizing to the parotid gland. PMID- 17398348 TI - Innominate artery hemorrhage following tracheostomy. AB - OBJECTIVES: To review the clinical presentation, predisposing factors, prevention strategies, management, and outcomes of innominate artery hemorrhage following tracheostomy. STUDY DESIGN AND SETTING: We report the case of an 80-y-old patient who experienced sudden massive innominate artery hemorrhage 11 days post tracheostomy. We review the literature and present recommendations for management and prevention. RESULTS: Emergency median sternotomy with ligation and resection of the affected segment was performed with no neurological or vascular sequelae. CONCLUSIONS AND SIGNIFICANCE: The risk of innominate artery hemorrhage may be minimized with simple measures. Management by ligation and resection of the pathological segment of artery has superior outcomes to primary vascular reconstruction with maintenance of blood flow. Bypass procedures are not routinely required and have not been shown to confer any significant benefit. Resection without reconstruction is associated with a surprisingly low incidence of neurological sequelae. PMID- 17398349 TI - Tracheo-innominate artery fistula: massive hemorrhage after decannulation. PMID- 17398350 TI - [Proceedings of the 7th National Infectious Disease (JNI) Meeting, Bordeaux, France, June 7, 8, 9th 2006]. PMID- 17398351 TI - Leadership. PMID- 17398352 TI - The "ulnar fovea sign" for defining ulnar wrist pain: an analysis of sensitivity and specificity. AB - PURPOSE: Eliciting tenderness in the region of the ulnar fovea is a possibly useful clinical test for defining the source of ulnar-sided wrist pain. Until now, no reports of the clinical sensitivity and specificity of this test have been available. Based on anecdotal observations, a hypothesis was developed stating that ulnar fovea tenderness (positive "ulnar fovea sign") is sensitive and specific in detecting two ulnar-sided wrist conditions: foveal disruption of the distal radioulnar ligaments and ulnotriquetral (UT) ligament injuries. METHODS: The clinical records of 272 consecutive patients with wrist arthroscopy performed by the senior author from 1998 through to 2005 were reviewed. Relevant clinical and surgical data were abstracted. The ulnar fovea sign test is executed by pressing the examiner's thumb distally into the interval between the ulnar styloid process and flexor carpi ulnaris tendon, between the volar surface of the ulnar head and the pisiform. A positive ulnar fovea sign is designated when there is exquisite tenderness that the patient claims replicates their pain, with comparisons made with the contralateral side. RESULTS: There were a total of 90 foveal disruptions and 68 UT ligament injuries diagnosed during wrist arthroscopy. The ulnar fovea sign was positive in 156 patients. The sensitivity of the fovea sign in detecting foveal disruptions and/or UT ligament injuries was 95.2%. Its specificity was 86.5%. CONCLUSIONS: The hypothesis stating that the ulnar fovea sign is a useful clinical maneuver to detect foveal disruptions and UT ligament tears is supported. The conditions represent 2 common sources of ulnar-sided wrist pain. The differentiation between the 2 conditions may be made clinically, where UT ligament tears are typically associated with a stable distal radioulnar joint and foveal disruptions are typically associated with an unstable distal radioulnar joint. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic II. PMID- 17398353 TI - Repair of the triangular fibrocartilage complex after ulnar-shortening osteotomy: second-look arthroscopy. AB - PURPOSE: Ulnar shortening is a widely used procedure for various conditions associated with ulnar wrist pain, including triangular fibrocartilage complex (TFCC) injury; however, few reports have examined the condition of the TFCC after osteotomy. The central avascular zone of the TFCC generally is considered to have no potential to heal. This study investigated whether the avascular zone of the TFCC has any potential for repair, and whether repair of the torn disc proper correlates with clinical findings. METHODS: Between 1987 and 2005, we performed 75 second-look arthroscopies after an ulnar-shortening osteotomy for ulnar wrist disorders. Of these, 32 wrists with a TFCC (disc proper) tear on first arthroscopy were included in this study. Data from patient charts, radiography, and video images of arthroscopy were reviewed retrospectively. Tears of the disc proper were classified as radial, central, or ulnar tears, and as either linear or round tears. RESULTS: Meticulous second-look arthroscopy showed repair of tears in 50% of studied wrists. Round tears tended to repair better than linear tears. Although the final clinical score was better in repaired wrists than in nonrepaired wrists, no marked differences were noted between groups in terms of age, gender, preoperative ulnar variance, follow-up period, or surgical procedures used. CONCLUSIONS: The avascular zone of the TFCC possesses some potential for repair; however, factors promoting spontaneous repair of this tissue were not identified. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 17398354 TI - In vivo three-dimensional displacement of the distal radioulnar joint during resisted forearm rotation. AB - PURPOSE: The in vivo displacement of the distal radioulnar joint (DRUJ) while under resisted pronation or supination has never been previously quantified. Being a joint with 3 dimensions of motion (dorsopalmar translation, changes in ulnar variance, and radioulnar coaptation), our purpose was to determine its 3 dimensional (3D) displacement during resisted rotational torque. METHODS: Ten normal subjects (aged 31.8 years+/-7.8; F, 5; M, 5) were recruited. Computed tomography scans of a subjects' distal forearms were obtained while grasping the handles of a custom apparatus to keep the fist and forearm in neutral rotation. Scanning was performed in 3 conditions: no rotational torque (control), maximum isometric supination, and maximum isometric pronation. The 3D displacement of the DRUJ, using the ulnar fovea as the reference point, during isometric rotational torque was determined using markerless bone registration techniques. RESULTS: The mean 3D displacement of the ulnar fovea during maximum isometric pronation and maximum isometric supination was 3.09 mm+/-0.94 and 2.17 mm+/-1.55, respectively. During maximum isometric pronation, the ulnar fovea displaced a mean of 0.51 mm+/ 1.79 dorsally. During maximum isometric supination, the ulnar fovea displaced a mean of 0.13 mm+/-2.07 volarly. In both conditions, ulnar variance increased (0.23 mm+/-2.52 for isometric pronation and 0.44 mm+/-1.57 for isometric supination), and there was increased radioulnar coaptation (0.37 mm+/-0.86 for isometric pronation, 0.039 mm+/-0.66 for isometric supination). CONCLUSIONS: Resisted rotational torque can be applied to the DRUJ while in neutral rotation during computed tomography imaging, producing measurable displacements in 3D. This technique of studying the DRUJ has the potential to improve our ability to detect and quantify instabilities of the DRUJ. PMID- 17398355 TI - Osteoarthritis of the thumb carpometacarpal joint in women and occupational risk factors: a case-control study. AB - PURPOSE: Among other etiologic factors involved in first carpometacarpal (CMC) osteoarthritis (OA), occupational factors have been postulated as influencing the occurrence of this condition. Very few epidemiologic studies, however, have evaluated this topic. Determining the occupational risk factors is important in proposing preventive measures at the workplace. This case-control study was undertaken to explore whether there was a history of greater exposure to some occupational factors (eg, occupations, hand postures, tasks involving the CMC joint) in women requiring surgery for CMC OA compared with women with no CMC OA noted by history and physical examination. METHODS: The case subjects were 61 women surgically treated for primary CMC OA and the control subjects were 120 aged matched women without history or features of CMC OA. A detailed structured interview was developed to elicit information about age, smoking habits, medical history, lifestyle history, and occupational factors. Occupational factors were based on a detailed history of jobs, coded according to the International Standard Classification of Occupations. For the main occupation/job held for the longest duration and during an average working day, subjects were asked about hand posture or tasks involving requirements presumed to cause a strain or a high load to the CMC joint and about certain work conditions. RESULTS: Of the 61 case and 120 control subjects, 5 and 14, respectively, had never worked. There was no difference between the average number of jobs through the working lifetime of the group of case subjects compared with the group of control subjects. Logistic regression analysis showed that after adjustment for age, smoking status, obesity, CMC OA family history, hysterectomy history, parity, and occasional job, the following occupational factors were risk factors for CMC OA: occupations presumed to be associated with increased risk for CMC OA, occupations involving repetitive thumb use, and jobs perceived by the subject having not enough rest breaks during a day. The group of case subjects had a higher prevalence of hysterectomy history and family CMC OA history compared with the group of control subjects. CONCLUSIONS: Although previous studies have reported that work and exposure history may lack precision as risk factors, our results give further evidence to support the role of certain occupational factors in the occurrence of CMC OA in women. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic II. PMID- 17398356 TI - Distribution of nerve endings in the human dorsal radiocarpal ligament. AB - PURPOSE: The purpose of this investigation was to evaluate the nerve-ending apparatus populations within a large number of adult human dorsal radiocarpal (DRC) ligaments to test the hypothesis that the majority of nerve endings could be grouped into established classifications and that the nerve endings could be found in predictable locations within the substance of the ligament. METHODS: The DRC ligaments were harvested from 20 wrists of 10 fresh cadavers with an average age of 75.6 years within 12 to 18 hours of death. Before the tissues were harvested, radiographs were taken to exclude any arthritic conditions of the wrists. Tissues were fixed, sectioned with a cryostat, and serial sections were collected on glass slides. Slides were processed for fluorescence immunohistochemistry using antibody to protein gene product 9.5 and a secondary antibody conjugated to a fluorescent tag (Alexa Fluor 488). The sections were evaluated with an LSM-510 confocal laser microscope and a Kontron KS 400 image analyzer. Labeled nerve endings were counted, mapped, and reconstructed. RESULTS: The average number of nerve endings in each DRC ligament was 10.1+/-4.7. More than 76% of the nerve endings were found in the 2 ends of the ligament with 23.3% in the central third and approximately 80% distributed in the superficial layer. More than 80% of the nerve endings were discovered in the epiligamentous sheath rather than in the perifascicular spaces. CONCLUSIONS: The distribution of the nerve endings follows a consistent pattern. These results will provide a foundation of morphologic information useful in understanding normal and abnormal neural control of wrist joint mechanics. PMID- 17398357 TI - Biomechanics and histology of intact and repaired digital nerves: an in vitro study. AB - PURPOSE: To investigate the biomechanical properties of intact and repaired cadaver digital nerves. METHODS: Ultimate tensile failure strength and stiffness were determined in 67 human cadaver digital nerves. Total nerve area, fascicular area, and nonfascicular (connective tissue) area were determined from the metacarpophalangeal to the distal interphalangeal joint in another 35 axial nerve sections to determine regional anatomic variation. Thirty-eight additional digital nerves were transected, and epineural repairs were performed using simple, interrupted sutures. Suture number (2 vs 4), gauge (8-0 vs 9-0), and purchase length (1 mm vs 2 mm) were used in various combinations, and then the repaired nerves were pulled to failure. The mechanism of repair-site failure was determined for each suture. In situ tension of the intact digital nerves was measured during passive metacarpophalangeal and proximal interphalangeal joint motion in another 19 intact digital nerves. RESULTS: There were no significant differences in failure load or stiffness with respect to the radial or ulnar nerves within a finger or between fingers. The primary tactile side of the finger tended to have a larger diameter digital nerve. Digital nerve failure was more common proximally than distally. Intact digital nerves failed at 6 N with a stiffness of 1 N/mm. Histologic analysis showed that fascicular area and total area decreased from proximal to distal, whereas the nonfascicular-to-fascicular area ratio increased. Four epineural sutures were statistically stronger than 2 sutures. Suture purchase length and gauge did not affect repair strength. The 8-0 nylon sutures failed primarily by cut-out, whereas the 9-0 sutures failed by cut out or breakage. Repaired nerves failed at 1 to 2 N. Maximal metacarpophalangeal joint hyperextension resulted in 4 N of digital nerve tension. When the metacarpophalangeal joint was not hyperextended, proximal interphalangeal joint motion did not generate tension. CONCLUSIONS: Similar to flexor tendons, the number of suture strands crossing the repair site was the most important variable affecting digital nerve repair strength in this cadaveric model. PMID- 17398358 TI - Perceptions of people with tetraplegia regarding surgery to improve upper extremity function. AB - PURPOSE: In the United States, more than 100,000 Americans live with the disability of tetraplegia. These individuals must struggle through long and complicated rehabilitations. Upper-extremity reconstructive surgery can improve use of the upper limb for appropriate candidates; however, a prior national study showed that these procedures rarely are performed. This cross-sectional survey identified the attitudes and beliefs of people with tetraplegia that may dissuade potential candidates from receiving these procedures. METHODS: An oral survey was designed to determine priorities of reconstruction in individuals with tetraplegia. This survey was administered to 50 people with tetraplegia. RESULTS: Among those surveyed, 13 (26%) had never heard of upper-extremity reconstructive surgery, but 22 (44%) were interested in upper-extremity reconstruction. People with tetraplegia who had a negative first impression of these procedures were far less likely to want reconstruction 0 (0%) vs. 11 (45%). Of patients who learned about these procedures from their physicians, 10 (67%) had a negative first impression after the physician consultation. CONCLUSIONS: Although many people with tetraplegia understand the benefits of upper-extremity reconstruction, a large number of them are unaware of or have unfavorable attitudes toward these procedures. These negative attitudes may account for the marked underuse of upper extremity reconstructive procedures in the United States. PMID- 17398359 TI - Clinical results of absorbable plates for displaced metacarpal fractures. AB - PURPOSE: To present the clinical results of a study of unstable metacarpal fractures treated with absorbable plates. METHODS: Between July 2004 and June 2006, 12 patients (14 fractures) who presented with displaced, unstable, metacarpal fractures had open reduction and internal fixation. The overall clinical follow-up results and radiographic controls at 6, 12, and 26 weeks after surgery are reported. The clinical outcome was assessed by the Disabilities of the Arm, Shoulder, and Hand score and the visual analogue pain scale. RESULTS: The involved fingers showed an average final total active motion of 234 degrees (range, 220 degrees-265 degrees). No deformity of rotation>5 degrees was observed in any patient in the clinical follow-up evaluation. One patient had a loss of reduction with a palmar angle of the metacarpal axis of 20 degrees in the sagittal plane. A second patient with secondary loss of reduction required surgical revision, at which time internal fixation was performed by using a titanium plate. Complications included keloid formation and prolonged soft-tissue swelling for more than 6 weeks in 3 patients. No wound margin necrosis, infection, pseudarthrosis, sinus formation, or osteolysis was observed. Bone consolidation was achieved reliably within 6 weeks both clinically and radiologically. The Disabilities of the Arm, Shoulder, and Hand score results 6 weeks after surgery were an average of 30 points compared with 13 after 12 weeks and 3 points after 26 weeks. The visual pain scale showed mean values of 18 after 6 weeks, 2 after 12 weeks, and 0.2 after 26 weeks. CONCLUSIONS: Absorbable plates are suitable for use in hand surgery and allow early range of motion in combination with an additional orthosis of the hand (as described) for 3 weeks. In the early and medium-term postoperative course, no osteolysis or sterile sinus formation was observed. Metal plates are still the gold standard; however, surgical implants made of amorphous copolymer of L-lactide and glycolic acid in combination with an orthosis constitute a useful and reliable means of metacarpal fracture treatment. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 17398360 TI - Osteolysis after the use of a silicon-stabilized tricalcium phosphate-based bone substitute in a radius fracture: a case report. AB - Skelite, a silicon-stabilized tricalcium phosphate-based bone substitute, is a synthetic alternative to the autogenous bone graft. We present a foreign body inflammatory reaction resulting in extensive osteolysis that occurred after use of Skelite as a void filler in the surgical reconstruction of a distal radius fracture. PMID- 17398361 TI - Fluoroscopic and magnetic resonance cross-sectional imaging assessments of radial and ulnar torsion profiles in volunteers. AB - PURPOSE: Planning an osteotomy to correct rotational malunions of the forearm is difficult because the uninvolved side is the only available reference to assess radial and ulnar torsions. This study was designed to compare the reliability of 2 methods for the determination of the torsion profile of both forearm bones and to assess side differences further in volunteers. METHODS: Fluoroscopy in combination with goniometry and magnetic resonance (MR) cross-sectional imaging were used to determine torsion profiles of the radius and the ulna in 24 asymptomatic volunteers. Interrater and interside reliabilities were assessed. RESULTS: For the radius, interclass correlation coefficients were less than 0.65 with fluoroscopy and greater than 0.80 with magnetic resonance imaging (MRI). For the ulna, both methods had an interclass correlation coefficient of greater than 0.90. Maximum side-to-side differences assessed with fluoroscopy and MRI were 25 degrees and 34.5 degrees for the radius and 20 degrees and 32 degrees for the ulna, respectively. There were no statistical differences between sides using both methods for both forearm bones. CONCLUSIONS: Fluoroscopy coupled with goniometry is a valuable method for assessing the torsion profile of the ulna. MR cross-sectional imaging is better to assess the torsion profile of the radius; however, a side difference in torsion profile of up to 35 degrees for the radius and of up to 20 degrees for the ulna should be considered physiologic. Hence, only side differences greater than these limits may serve as an indication for an axial osteotomy in the clinical setting. PMID- 17398362 TI - The strength of the 6-strand modified Kessler repair performed with triple stranded or triple-stranded bound suture in a porcine extensor tendon model: an ex vivo study. AB - PURPOSE: To investigate the biomechanic influence of triple-stranded sutures and the spatial arrangement of the strands on the strength of the 6-strand Pennington modified Kessler repair. METHODS: In the present ex vivo study of pig extensor tendons 2 techniques were used: (1) triple-stranded suture (3 suture strands in the same needle) and (2) triple-stranded bound suture (3 suture strands in the same needle that were bound together, parallel to each other, side by side). The repairs were subjected to static tensile testing. RESULTS: The 6-strand modified Kessler repair performed with triple-stranded bound suture reached significantly higher yield force, ultimate force, and both partial and total 1-, 2-, and 3-mm gap forces compared with the repairs performed with triple-stranded suture. The stiffness and strain values at the yield point and at the ultimate point did not differ significantly. CONCLUSIONS: This experimental study introduces a way to improve the strength of the tendon repair. The triple-stranded bound suture significantly increased both the gap resistance and ultimate force of the 6 strand modified Kessler repair. We assume the improvements are due to increased holding capacity of the locking loops. The triple-stranded bound suture is easy to use and avoids several problems associated with traditional multistrand repairs. Further studies are needed before clinical use can be considered. PMID- 17398363 TI - An analysis of factors associated with failure of tendon repair in the canine model. AB - PURPOSE: The canine model is commonly used for flexor tendon repair research. The purpose of this study was to analyze the factors, including laceration mode (partial and complete), suture techniques, therapy methods, and weight-bearing status, associated with tendon repair rupture or gap formation in the canine model in vivo. METHODS: We reviewed the factors associated with repair failure among 624 flexor tendon repairs in zone II from 242 dogs reported previously from our institution, including both partial and complete lacerations. RESULTS: We found that weight-bearing due to failure of postoperative immobilization was the most important factor influencing tendon repair rupture or gap formation. CONCLUSIONS: As has been noted clinically, in our canine model failure and gapping of a flexor tendon repair was primarily the result of uncontrolled loading. Rehabilitation strategies that reduce the risk of catastrophic loading of the repair are critical to reducing the experimental failure rate when using dogs for flexor tendon research. Similar strategies may also reduce such failures in humans. PMID- 17398364 TI - Functional deficit after transfer of the pronator teres for acquired radial nerve palsy. AB - PURPOSE: A suitable muscle motor in reconstruction after acquired never injury should have adequate strength to perform the desired task, be aligned in the direction of pull, have synergistic action, and not result in unacceptable functional loss. In radial nerve palsy, the pronator teres is the most common motor donor used to restore wrist extension. Although the pronator teres remains aligned to provide pronation, the force deficit of the transfer is not known. METHODS: We used 6 cadavers and 6 patients to determine the loss of pronation strength both experimentally and clinically. RESULTS: Cadaveric testing showed a loss of pronation produced with similar load after transfer of the pronator. Clinical testing showed statistically significant loss of pronation ranging from 24% to 44%, depending on the method of testing. This deficit may be an important consideration in some clinical situations when transfers are used while waiting for radial nerve function to return. CONCLUSIONS: In the cadaveric biomechanic testing, we simulated the pronator teres-to-extensor carpi radialis brevis tendon transfer and showed a decreased range of motion and force developed after transfer. The clinical arm of the study confirmed our biomechanic findings by showing the loss of pronation function. This loss may be an important factor when planning reconstruction for radial nerve injuries. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV. PMID- 17398365 TI - Comparison of instruments to assess hand function in children with radius deficiencies. AB - PURPOSE: For treatment evaluation of children with radius deficiencies (RDs), standardized assessment of hand function in performing activities is required. Instruments to measure hand function have been developed for other diagnoses. The current study aimed to find additional evidence for validity, reliability, and usefulness of these instruments for children with RDs. METHODS: In this study, 20 children with RDs (aged 4-12 years) participated; 16 were boys, and 13 children were unilaterally affected. Children were assessed using the Assisting Hand Assessment, the Unilateral Below Elbow Test, the Prosthetic Upper Extremity Functional Index, and ABILHAND-Kids. Construct and convergent validity of the instruments were studied focusing on predefined hypotheses and relationships with other instruments and the therapist's global assessment. Test-retest reliability was assessed in 10 children by means of the intraclass correlation coefficients and the smallest detectable differences. RESULTS: For children with RDs, the Assisting Hand Assessment and the Prosthetic Upper Extremity Functional Index appeared to be the most valid function test and questionnaire according to the relationships found with type of RD, functional hand grips, and the therapist's global assessment of hand function. Regarding test-retest reliability, intraclass correlation coefficients ranged from 0.82 to 0.91, and smallest detectable differences were acceptably small. CONCLUSIONS: The current results contribute to the evidence that the instruments, especially the Assisting Hand Assessment and the Prosthetic Upper Extremity Functional Index, provide valid and reliable results in children with RDs. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic I. PMID- 17398366 TI - Current practice of microsurgery by members of the American Society for Surgery of the Hand. AB - PURPOSE: First, to determine the percentage of members of the American Society for Surgery of the Hand (ASSH) that use microsurgical techniques as part of their surgery practice, and second, to identify factors limiting their use of these techniques. METHODS: A 34-item, anonymous, Web-based survey was sent to all active ASSH members. Twelve items concerned demographics and 22 items addressed prior microsurgical training, current use of these techniques, factors currently limiting their use of these techniques, and potential methods to address these limiting factors. RESULTS: Responses were received from 561 of 1,238 of the ASSH members contacted (45% response rate). Most had residency training in orthopedics (N=460, 82%) or plastic surgery (N=79, 14%), followed by a hand fellowship in an orthopedic (N=363, 62%) or combined program (N=170, 30%). More than 54% (N=304) practiced privately, 33% (N=184) practiced in tertiary institutions, and the remainder practiced at regional centers. Of those responding, 505 (90%) stated that hand surgery constituted more than 50% of their practice, whereas for 527 (94%) respondents microsurgery comprised less than 25%. Most members (N=398, 71%) accepted emergency patients, of which 223 (56%) at a referral center. Three hundred sixteen respondents (56%) performed replantations, of whom 196 (62%) performed fewer than 5 per year. Four hundred fifteen respondents (74%) observed a decrease in replantation attempts over the past decade. This was attributed to refinement of indications (N=17, 83%), fewer patients with amputations (N=116, 28%), and declining reimbursement (N=344, 4%). Reasons for not personally performing replantations included busy elective schedules (N=125, 51%), inadequate confidence in performing replantations (N=96, 39%), and disappointment in results (N=56, 23%). Thirty percent (N=74) stated they would reconsider performing replantations if reimbursement was greater. Practice rates of examined microsurgical procedures ranged from 22% to 57%, although most had received microsurgical training. Despite rating their fellowship as excellent (N=393, 70%) or good (N=135, 24%), only 315 (56%) considered their present microsurgical skills to be above average. Many respondents believed that they would benefit from continuous training through continuing education courses. CONCLUSIONS: Educational, economic, and practical factors discourage the clinical application of microsurgical technique by hand surgeons. This unfavorable environment should be addressed by policy-making organizations and continuous surgical training. TYPE OF STUDY/LEVEL OF EVIDENCE: Other/Survey. PMID- 17398367 TI - Fingertip replantation. AB - Fingertip replantation is now an established technique. Although successful replantation is an ideal method for treatment of fingertip amputation, various other methods still are widely used and may be functionally acceptable. The indications for replantation to treat fingertip amputation is still controversial. This article presents a global view of the current status of replantation for the treatment of fingertip amputation. The surgical technique, strategies to overcome postoperative congestion, and overall results are discussed. PMID- 17398368 TI - The correction of postburn contractures of the second through fourth web spaces. AB - The release of postburn contractures in the hand is one of the most commonly performed procedures in burn injuries. Contractures of the web space may involve palmar, dorsal, or both sides of the web skin and require various surgical techniques. In this report we provide general guidelines for the release of these contractures, with special emphasis on reconstruction of the second through fourth web spaces. PMID- 17398369 TI - First rib resection in thoracic outlet syndrome. AB - Most patients with thoracic outlet syndrome (TOS) present with exercise-induced upper extremity paresthesia. Neurogenic TOS is the most common type where the brachial nerve plexus is compressed against a tight thoracic outlet. Vascular compromise although rare can result from thoracic outlet pressure against the subclavian artery or more commonly the subclavian vein. This article reviews the pathophysiology of TOS and describes several effective surgical interventions. Complete first rib resection with surgical decompression is an essential part of the treatment for TOS. First rib resection via supraclavicular or a preferred transaxillary route should be considered when conservative modalities provide no symptom improvement. PMID- 17398371 TI - History of distal ulna resection. PMID- 17398372 TI - Randomized trial of the effects of cholesterol-lowering with simvastatin on peripheral vascular and other major vascular outcomes in 20,536 people with peripheral arterial disease and other high-risk conditions. AB - OBJECTIVES: The Heart Protection Study (HPS) provides an opportunity to assess directly the effects of cholesterol-lowering therapy on major vascular events (defined as myocardial infarction, coronary death, stroke, or revascularization) in patients with peripheral arterial disease (PAD). In addition, the effects on peripheral vascular events (ie, non-coronary revascularization, aneurysm repairs, major amputations or PAD deaths) can be assessed. METHODS: 6748 UK adults with PAD and 13,788 other high-risk participants were randomly allocated to receive 40 mg simvastatin daily or matching placebo, yielding an average LDL cholesterol difference of 1.0 mmol/L (39 mg/dL) during a mean of 5 years. RESULTS: For participants with PAD, allocation to simvastatin was associated with a highly significant 22% (95% CI 15-29) relative reduction in the rate of first major vascular event following randomisation (895 [26.4%] simvastatin-allocated vs 1101 [32.7%] placebo-allocated; P < .0001), which was similar to that seen among the other high-risk participants. The absolute reduction in first major vascular event was 63 (SE 11) per 1000 patients with PAD and 50 (SE 7) per 1000 without pre-existing PAD. Overall, among all participants, there was a 16% (5-25) relative reduction in the rate of first peripheral vascular event following randomisation (479 [4.7%] simvastatin vs 561 [5.5%] placebo), largely irrespective of baseline LDL cholesterol and other factors. This effect chiefly reflects a 20% (8-31) relative reduction in non-coronary revascularization procedures (334 [3.3%] vs 415 [4.0%]; P = .002). CONCLUSION: HPS demonstrates the benefits of cholesterol-lowering statin therapy in patients with PAD, regardless of their presenting cholesterol levels and other presenting features. Allocation to 40 mg simvastatin daily reduces the rate of first major vascular events by about one-quarter, and that of peripheral vascular events by about one-sixth, with large absolute benefits seen in participants with PAD because of their high vascular risk. Consequently, statin therapy should be considered routinely for all patients with PAD. PMID- 17398375 TI - Contemporary results of open repair of ruptured descending thoracic and thoracoabdominal aortic aneurysms. AB - OBJECTIVE: The purpose of this study was to evaluate the results of open repair for ruptured descending thoracic and thoracoabdominal aortic aneurysm (RDTAA). METHODS: A retrospective review identified 41 consecutive cases of open surgical repair in 40 patients presenting with nontraumatic, atherosclerotic RDTAA from 1996 to 2006. Patients with traumatic injuries or complicated dissections were excluded. Patient characteristics and preoperative, intraoperative, and postoperative variables were collected from the medical record. Univariate and logistic regression were used to identify factors contributing to mortality and morbidity in these patients. RESULTS: The operative mortality rate was 26.8% (11/41). All but two deaths occurred within 24 hours of operation; seven were intraoperative. Overall actuarial survival rates at 1 and 2 years were 53.7% and 47.1%, respectively. For those who survived to hospital discharge, the respective numbers were 73.3% and 64.4%. Intraoperative hypotension and blood transfusion requirements were independent predictors of perioperative death. Octogenarians had a mortality rate equivalent to that of the younger population (25% vs 27.6%; not significant). There was a strong trend toward an improved outcome in the latter part (2003-2006) compared with the first part (1995-2002; 13.6% vs 42.1%, respectively; P = .075). CONCLUSIONS: Direct open repair for RDTAA can be achieved with acceptable mortality and morbidity rates even in elderly patients. Improved outcome can be expected with increased volume and experience. This series should help establish a reference against which the results of endovascular endeavors and hybrid procedures could be compared. PMID- 17398377 TI - Suprarenal fixation of endovascular aortic stent grafts: assessment of medium term to long-term renal function by analysis of juxtarenal stent morphology. AB - OBJECTIVE: The effect of suprarenal stent fixation during endovascular aortic aneurysm repair (EVAR) on renal function remains unclear. A unique validated three-dimensional intraluminal imaging technique was used to analyze the effect of suprarenal stent position relative to renal artery orifices. Also analyzed was its medium-term to long-term effects on renal function. METHODS: The study cohort comprised 29 of 34 consecutive patients who underwent EVAR with the Zenith endograft system from September 1999 to March 2002 at a single institution. The precise locations of the uncovered suprarenal stent struts were assessed by a virtual endoscopic imaging technique. Anatomic and quantitative categorization of patients was made according to the configuration of uncovered stent struts across the renal artery ostia (RAO). The anatomic subgroups were defined as struts located centrally or peripherally across both RAO. The quantitative subgroups were defined as RAO crossed by multiple struts, a solitary strut, or no struts. The subgroups were compared for their renal function, as assessed by temporal measurements of serum creatinine concentration and creatinine clearance, and renal parenchymal perfusion defects, as assessed using contrast-enhanced computed tomography (CT). RESULTS: Mean follow-up was 52.7 months. Separate subgroup analysis for both anatomic and quantitative configurations did not demonstrate any significant difference in renal function between the different strut permutations (P > .05). Follow-up imaging confirmed one case of renal infarction secondary to an occluded accessory renal artery, although this patient had normal serum creatinine levels. CONCLUSION: RAO coverage by suprarenal uncovered stents does not appear to have a significant effect on renal function as evaluated by creatinine measurements in patients with normal preoperative renal function. PMID- 17398378 TI - Low vitamin B6, and not plasma homocysteine concentration, as risk factor for abdominal aortic aneurysm: a retrospective case-control study. AB - BACKGROUND: Hyperhomocysteinemia has been associated with vascular disease in many epidemiologic studies, but only a few have reported on the relation between hyperhomocysteinemia and aneurysms of the abdominal aorta (AAAs). Although these studies showed higher homocysteine concentrations in patients with AAA than in controls, little attention had been given to possible confounding factors. Most patients with AAA are of older age, have an impaired renal function, and have other risk factors for cardiovascular disease. This matched case-control study investigated the relation between homocysteine concentration (before and after methionine loading) and AAA, taking into account possible confounders such as age, sex, and concentrations of creatinine and B vitamins. METHODS: Patients with a history of AAA were recruited from the outpatient clinic; 60% had already undergone surgery for their AAA. They were asked to invite a friend or neighbor to participate as a control subject (age-matched and sex-matched). Concentrations of homocysteine, vitamin B6, vitamin B12, folate, and creatinine were determined in the fasting state, and blood was taken for methylenetetrahydrofolate reductase (MTHFR) mutation analysis. Six hours after oral methionine loading, the postmethionine load homocysteine concentration was determined. RESULTS: Univariate analysis showed an odds ratio (OR) of 2.2 (95% confidence interval (CI), 0.9 to 5.5) for the risk of AAA for the highest quartile of homocysteine concentration. After adjustment for creatinine, the OR was markedly reduced to 1.24 (95% CI, 0.42 to 3.66), and this risk further attenuated in the multivariate analysis. Univariate analysis of the B vitamins showed an increased risk of AAA for the bottom quartile of vitamin B6 (OR, 3.75; 95% CI, 1.22 to 11.54), which even increased after adjustments. The relative risk associated with the MTHFR 677TT polymorphism was 2.1 (95% CI, 0.9 to 5.3). CONCLUSION: Vitamin B6, but not homocysteine, is an independent risk factor for AAA. The role of vitamin B6 in the pathogenesis of AAA needs to be further elucidated. PMID- 17398379 TI - Early complications and long-term outcome after open surgical treatment of popliteal artery aneurysms: is exclusion with saphenous vein bypass still the gold standard? AB - BACKGROUND: Popliteal artery aneurysms (PAAs) are rare, but thromboembolic complications may result in limb loss. To define complications and outcomes after open surgical repairs, we reviewed our experience. METHODS: Clinical data of patients with PAA seen between 1985 and 2004 at Mayo Clinic, Rochester, Minnesota, were reviewed and outcomes in 289 patients with open revascularization were analyzed. Kaplan-Meier method with log-rank tests, chi(2), and Wilcoxon rank sum tests were used for analysis. RESULTS: A total of 358 PAAs were treated in 289 patients, consisting of 281 (97%) men and eight (3%) women. There were 133 (46%) unilateral and 156 (54%) bilateral PAAs with a mean diameter of 2.9 cm (range, 1.5 to 9 cm). Abdominal aortic aneurysm (AAA) was more frequent with bilateral than unilateral PAAs (65% [101/156] vs 42% [56/133] P = .001). There were 144 (40%) asymptomatic limbs (group 1), 140 (39%) had chronic symptoms (group 2), and 74 (21%) had acute ischemia (group 3). Great saphenous vein (GSV) was used in 242 limbs (68%), polytetrafluoroethylene (PTFE) in 94 (26%), and other types of graft in 22 (6%). Early mortality was 1% (3/358), all in group 3 (4% [3/74]). Six of seven patients with perioperative myocardial infarctions belonged to group 3 (8%). The 30-day graft thrombosis rate was 4%, with 1% in group 1 (1/144), 4% in group 2 (5/140), and 9% in group 3 (7/74). All six early amputations (8%) were in group 3, five with failed bypass (4 PTFE, 1 GSV). Mean follow-up was 4.2 years (range, 1 month to 20.7 years). The 5-year primary and secondary patency rates were 76% and 87%, respectively, higher with GSVs (85% and 94%) than PTFE (50% and 63%, P < .05). Seven recurrent PAAs (2%) required reintervention. The 5-year freedom from reintervention was 100% after endoaneurysmorrhaphy vs 97% after ligations (P = .03). Five-year limb salvage rate was 97% (85% in group 3). There was no limb loss in group 1 and none in group 2 with GSV. In group 3, preoperative thrombolysis reduced the amputation rate in class II patients with marginally threatened limbs (96% vs 69%, P = .02). CONCLUSION: Acute presentation of PAA continues to carry high mortality and cardiac morbidity; although preoperative thrombolysis appears to improve results, the 8% early and 15% late amputation rates remain ominous. Early elective repair is recommended because these patients had no surgical mortality, a low rate of complications, and asymptomatic patients had no limb loss at 5 years. GSV and endoaneurysmorrhaphy continues to be the gold standard for open repair of PAA. PMID- 17398382 TI - Renal artery stenting slows the rate of renal function decline. AB - OBJECTIVE: The primary objective of this study was to analyze renal artery interventions performed at a tertiary medical center and to evaluate improvements in hypertension and renal excretory function. METHODS: A retrospective analysis was performed of patients treated at a tertiary medical center from January 2001 to December 2005. All patients treated with renal artery stenting by the Interventional Radiology or Endovascular Services were included. Descriptive and inferential analyses were performed. RESULTS: Forty patients with renal artery stenosis were evaluated for renal artery stenting, of these 22 were followed up with medical management. Twenty-six renal artery stents were placed in 18 patients (mean age, 70 +/- 8 years), of whom 76% were treated for multidrug resistant hypertension, and 24% were treated for renal salvage. Mean follow-up was 15 months. Patients experienced a significant reduction in hypertension and in the number of antihypertensive agents, but this significance deteriorated at 6 months, when their blood pressure and number of medications returned to preprocedural values. Compared with a cohort that was followed up with medical management, the rate of renal function decline improved from -0.08 mg/dL per month to 0.00 mg/dL per month (P < .05) after intervention. Patients with baseline chronic renal insufficiency experienced the greatest benefit from renal artery stenting. CONCLUSIONS: Renal artery stenting initially improves hypertension control, but the durability is lost after 6 months. Renal artery stenting dramatically slows the rate of renal function decline and could potentially delay a patient's requirement for hemodialysis. PMID- 17398384 TI - Plasma creatine kinase indicates major amputation or limb preservation in acute lower limb ischemia. AB - OBJECTIVE: Acutely ischemic limbs are often of uncertain viability. To assist operative management, this study determined prospectively which indicators on admission were the best predictors of major amputation and, conversely, limb preservation. METHODS: Data were collected on admission. Presenting complaint, history, clinical assessment, and blood test results, including creatine kinase (CK), were recorded. Surgical procedures were noted-in particular, the presence or absence of major amputation by death or discharge. The setting was a tertiary vascular referral center in a university teaching hospital. Subjects included all patients referred as emergency cases to the vascular unit over an 18-month period who were admitted for inpatient management with acute lower limb ischemia. The main outcome measure was major amputation. RESULTS: A total of 97 patients with acute ischemia were studied prospectively (51 men and 46 women). Twenty-one patients (21.6%) underwent major amputation. Previous vascular surgery (P = .012), mottling (P = .001), sensory loss (P = .003), motor loss (P = .001), muscle tenderness (P < .001), absent ankle Doppler signals (P = .008), neutrophilia (P = .011), and increased CK (P < .001) were significantly associated with major amputation. If CK was normal, the risk of major amputation was 4.6% (95% confidence interval, 0.0%-9.7%). If CK was increased, the risk was 56.2% (95% CI, 39.1%-73.4%). CONCLUSIONS: Specific clinical findings were significantly associated with major amputation. Of these, only CK had a positive predictive value greater than 50%. Plasma CK can assist operative management of acute lower limb ischemia by quantifying prospectively the risk of major amputation or limb preservation on admission. PMID- 17398385 TI - Superficial femoral artery percutaneous intervention is an effective strategy to optimize inflow for distal origin bypass grafts. AB - BACKGROUND: Iliac angioplasty in preparation for an infrainguinal bypass graft has been shown to be an effective strategy. We undertook this study to determine if superficial femoral artery (SFA) angioplasty offers durable inflow for distal origin grafts originating from the SFA or popliteal artery. METHODS: All distal origin grafts performed at a single institution between 1988 and 2006, 55 kg/m(2), hypercoagulable state, severe immobility, venous stasis, or previous history of DVT or pulmonary embolus. Using both Site-Rite (Bard Access Systems, Salt Lake City, Utah) ultrasound and fluoroscopy, Gunther Tulip (Cook, Inc., Bloomington, Ind) rVCFs were placed immediately after general anesthesia, just preceding the bariatric procedure. The internal jugular vein was the preferred approach, followed by the femoral vein. Retrieval was performed after the fourth postoperative week. RESULTS: During a 16-month period, 60 rVCFs were placed in 61 attempts, 57 through the internal jugular vein and three through the femoral vein. Six patients refused the retrieval attempt. Of the remaining 54 rVCFs, the primary retrieval success was 90% (49/54), with all failures due to severe filter tilt. The secondary retrieval success was 100% (3/3). The two remaining patients refused secondary retrieval attempt. The mean +/- standard deviation dwell time of the rVCFs was 63 +/- 30 days. No procedure complications occurred in placement or retrieval. One patient developed a clinical pulmonary embolism with the filter in place while not receiving postoperative anticoagulation. No patients died. The mean body mass index of the patients was 61 +/- 10 kg/m(2). CONCLUSION: Placement and retrieval of retrievable vena cava filters in high-risk bariatric surgery patients is safe, feasible, and offers potential clinical benefit to patients requiring short-term protection from pulmonary embolism. PMID- 17398389 TI - Vena cava filters and inferior vena cava thrombosis. AB - OBJECTIVE: Retrievable vena cava filters (R-VCF) are a recent addition to the therapeutic armamentarium for the prevention of pulmonary embolism. However, unlike permanent vena cava filters (P-VCF), outcomes data are limited regarding complication rates. METHODS: This was a retrospective comparative analysis of consecutive patients undergoing placement of R-VCF vs P-VCF at Wake Forest University School of Medicine from January 2000 to December 2004. Data collected included demographics, procedural specifics, filter type, indications, and complications. Summary data are expressed as number (percentage) or mean +/- SD. Continuous and categorical variables were analyzed by using t and Fisher exact testing, as appropriate. Four additional patients with vena cava thrombosis were also referred to our institution for treatment during the study period, all with opposed biconical VCFs (OptEase and TrapEase filters) recently placed at other facilities. This last group of patients is described but not included in the analysis. RESULTS: A total of 189 VCF (165 P-VCF and 24 R-VCF) cases were examined. No significant differences in VCF groups were observed according to age, documented hypercoagulability, or concomitant anticoagulation. Significant differences were observed according to sex (30.3% of P-VCF vs 62.5% of R-VCF patients were female), morbid obesity (4.2% of P-VCF vs 25% of R-VCF patients), active malignancy (20% of P-VCF vs 41.7% of R-VCF patients), and indication for VCF placement. Over a median follow-up of 8.5 months, no case of significant hemorrhage, no VCF migration, and four cases of vena cava thrombosis were observed. Vena cava thrombosis was observed more frequently in the presence of R VCF when compared with P-VCF (12.5% vs 0.6%; P = .007). All observed vena cava thromboses were associated with severe clinical symptoms and occurred in patients who received opposed biconical VCF designs. CONCLUSIONS: In our experience, both P-VCF and R-VCF can be placed safely. Among both permanent and retrievable devices, however, opposed biconical designs seem to be associated with an increased risk for vena cava thrombosis. Although causative factors remain unclear, filter design and resultant flow dynamics may play an important role, because all episodes of vena cava thrombosis occurred in patients with a single filter design. PMID- 17398390 TI - Remodeling of experimental arteriovenous fistula with increased matrix metalloproteinase expression in rats. AB - OBJECTIVE: Venous dilatation and wall thickening are part of the maturation of an arteriovenous fistula (AVF). However, the underlying mechanism of AVF remodeling remains unknown. We therefore studied whether matrix remodeling elicited by matrix metalloproteinases (MMPs) may contribute to AVF maturation. METHODS: A femoral AVF model in rats was established by invagination of the distal end of the left femoral artery into the femoral vein after venotomy (fistula group). In the sham group, the left femoral artery was cut, but venous invagination was not performed. Changes in the hemodynamics and the diameter of the iliac vein were studied on days 3, 14, and 28, then the iliac vein was removed and examined for changes in wall thickness and expression of MMP-2 and MMP-9, type 4 tissue inhibitor of metalloproteinases (TIMP-4), and collagen I and III by immunohistochemical staining or Western blotting. RESULTS: Femoral AVF resulted in a sixfold increase in blood flow in the fistula iliac vein and a gradual, but significant, increase in the thickness of the intima and media and marked up regulation of MMP-2 and MMP-9, down-regulation of TIMP-4, as well as degradation of collagens I and III. The collagen I/III ratio was significantly higher in the 14-day fistula group (1.44 +/- 0.32) than in the sham group (0.82 +/- 0.15) and was even higher in the 28-day fistula group (1.76 +/- 0.21). CONCLUSION: The present results confirmed our hypothesis that a high blood flow rate in the fistula vein affects the expression of MMPs and TIMP-4, resulting in the remodeling or maturation of the AVF. Remodeling is associated with degradation of collagen, with an increase in the collagen I/III ratio. PMID- 17398391 TI - Development of a spontaneously beating vein by cardiomyocyte transplantation in the wall of the inferior vena cava in a rat: a pilot study. AB - PURPOSE: This study was conducted to determine whether it is feasible to develop a vein that rhythmically beats by implanting immature cardiomyocytes in its wall. METHODS: Neonatal cardiomyocytes (5 x 10(6) cells each) were transplanted into the wall of the inferior vena cava in six female Fischer rats; in six rats, only the medium was transplanted. At 3 weeks after transplantation, the grafted site of the inferior vena cava was exposed and videotaped, and then processed for histology. RESULTS: Distinct rhythmic beating of the vena cava at the site of cell injection (at a rate lower than aortic beating) was observed in all six rats treated with neonatal cardiomyocyte injections, but in none of the six that received the medium. The vena cava continued to beat spontaneously and rhythmically after the aortas were clamped and after the heart was excised. The beating was manifest by visual contraction and relaxation of the vessel wall. The spontaneous beating rate was 101 +/- 7 beats/min at 1 to 3 minutes after excision of the heart. Hematoxylin and eosin staining showed viable grafts in the wall of the vena cava in all that were implanted with neonatal cardiac cells; but in none of the vena cava that received the medium. Neonatal cardiomyocytes in the graft matured with cross striations and stained positive for the muscle marker sarcomeric actin. CONCLUSIONS: The present study demonstrates that neonatal cardiomyocytes survive, mature, and spontaneously and rhythmically contract when implanted in the wall of a vein. PMID- 17398392 TI - Comparison of the vascular responses to balloon-expandable stenting in the coronary and peripheral circulations: long-term results in an animal model using the TriMaxx stent. AB - OBJECTIVE: The clinical results after stenting in the coronary and peripheral circulations are vastly different. Possible explanations for this discrepancy include generally longer and more complex lesions in the periphery, variable vascular responses to injury according to anatomic location, disparate blood flow rates, and impedance in coronary vs skeletal smooth muscle beds, or phenotypic differences in neointimal hyperplasia and remodeling. This study examined the long-term results (6 months) after implantation of phosphorylcholine-coated balloon-expandable stents in a porcine model of experimental in-stent coronary and peripheral arterial restenosis. METHODS: Forty-eight stainless steel-tantalum stainless steel composite balloon-expandable stents coated with phosphorylcholine (TriMaxx stent, Abbott Laboratories, Abbott Park, Ill) were implanted in the coronary (3.0 x 15 mm) or larger femoral arteries (4.0 x 38 mm) of Yorkshire crossbred swine to achieve a 1.1:1 stent-to-artery ratio. After 28, 90, or 180 days, the arteries were excised, perfusion-fixed at 100 mm Hg, sectioned, and stained with hematoxylin and eosin for morphometric evaluation. RESULTS: One animal did not survive to euthanasia; all arteries in surviving animals were patent. No significant differences were found in mean injury or inflammation scores among the groups or time points. The larger femoral arteries generated more neointimal area over time than the coronary arteries. The neointimal area in coronary arteries was 2.76 +/- 0.71, 1.75 +/- 0.42, and 1.83 +/- 0.19 mm(2) at 28, 90, and 180 days, respectively, and 5.20 +/- 0.97, 3.11 +/- 0.53, and 5.10 +/ 0.80 mm(2) in the femoral arteries (P < .05 coronary vs femoral at 180 days). This led to statistically significantly increased percent area stenosis at 180 days (coronary 27% +/- 4% vs femoral 45% +/- 5%; P < .05). CONCLUSIONS: The vascular response to balloon-expandable stenting in the coronary and peripheral circulations is different. After 6 months, neointimal hyperplasia and stent induced stenosis were increased in peripheral porcine arteries compared with coronary arteries. PMID- 17398393 TI - Detection of aortic graft infection by 18-fluorodeoxyglucose positron emission tomography combined with computed tomography. AB - Functional in vivo molecular imaging is provided with 18-fluorodeoxyglucose positron emission tomography (FDG-PET), which can detect cells with high glucose turnover. FDG-PET is an established imaging tool in oncology but has also been used in infectious and inflammatory diseases. PET combined with computed tomography (PET/CT) shows the metabolic activity with precise anatomic localization. More than 2000 scanners have now been installed worldwide, and with better availability, this hybrid method has the potential to become an important imaging tool in the management of suspected aortic graft infections, especially in patients with low-grade graft infection. We report a patient with a suspected aortic graft infection that was confirmed and anatomically localized by FDG PET/CT. An extra-anatomic bypass and extirpation of the aortic graft was performed. The perioperative location of the graft infection coincided exactly with the place of FDG uptake shown on PET/CT. The patient had an uneventful postoperative recovery and did well during 6 months of follow-up. PMID- 17398394 TI - Recurrent spinal cord ischemia after endovascular stent grafting for chronic traumatic aneurysm of the aortic isthmus. AB - This report describes the development of recurrent spinal cord ischemia in a patient after insertion of a stent graft into the upper segment of the descending thoracic aorta for the treatment of a chronic traumatic aneurysm of the aortic isthmus. Intraoperatively, the stent covered the ostium of the left T7 artery, which was shown to give rise to a middle dorsal artery by postoperative spinal cord arteriography. PMID- 17398395 TI - Aortoduodenal fistula 5 years after endovascular abdominal aortic aneurysm repair with the Ancure stent graft. AB - We report a case of aortoduodenal fistula 5 years after uncomplicated endovascular abdominal aortic aneurysm repair. The diagnosis was confirmed by abdominal computed tomography scan and esophagogastroduodenoscopy. The patient was successfully treated with primary duodenal repair, removal of the infected graft, in situ placement of a bifurcated graft, and omental interposition. Review of the literature identifies this as one of very few documented aortoduodenal fistulas after endovascular aneurysm repair. Fistulization occurred despite accurate stent graft placement without migration, endoleak, or aortic sac size enlargement on annual postoperative imaging studies. PMID- 17398396 TI - The nutrient omentum free flap: revascularization with vein bypasses and greater omentum flap in severe arterial ulcers. AB - Microvascular transfer of the omentum provides well-vascularized and pliable tissue but has not widely been used in vascular extremity reconstruction because of the potentially high donor site morbidity caused by the necessary laparotomy. Laparoscopic minimally invasive harvest of a free greater omentum flap and microsurgical transfer of this tissue with split skin grafts on top and connected to sequential vein bypasses may be a an interesting new modality when other reconstructive options are absent or scarce. We report this novel technique in a patient with arterial occlusive disease and an extensive and circular ulcer of the lower extremity. PMID- 17398397 TI - True ulnar artery aneurysm of the hand in an 18-month-old boy: a case report. AB - True aneurysms of the hand arteries are rare and are exceedingly uncommon in children. Presented is a case of a true ulnar artery aneurysm in an 18-month-old boy in which there was no history of trauma. The aneurysm was resected without reconstruction because of the normal preoperative Allen test result, normal preoperative finger pressure measurement with ulnar artery occlusion, the angiographic evidence that the radial artery was the dominant artery of the hand, and intraoperative evidence of adequate hand perfusion after excluding the aneurysm from the hand circulation as documented by good Doppler signals in all digital arteries. PMID- 17398398 TI - Aneurysm of the inferior vena cava. PMID- 17398400 TI - Altering the career choice: can we attract more women to vascular surgery? AB - Vascular surgery has relied upon the pool of general surgical residents for candidates for residency. This has significantly limited the pool of applicants. With the advent of new primary certificate training paradigms, we investigated factors that might alter the career choices for women medical students. A Web based survey was conducted of third-year and fourth-year medical students at three university-affiliated programs. A total of 140 students completed the survey, which identified the third year of medical school as the primary year when students committed to a final career choice. Lifestyle issues were the primary deterrents, with long work hours and lack of time to see family negatively influencing students' interest in surgery. Eliminating the need for 5 years of general surgery and the opportunity for part-time training were cited as factors that would alter students' decisions to pursue vascular surgery. Ultimately, a positive rotation experience, lifestyle issues, and mentors impacted on final career decisions. To attract the best applicants to the field, we need to continue to be active in mentoring students and exploring alternative training paths and eventual career practices. PMID- 17398401 TI - Matrix metalloproteinases in peripheral vascular disease. AB - Matrix metalloproteinases (MMPs) are extracellular matrix-modifying enzymes that are important in many physiologic and pathologic vascular processes. Dysregulation of MMP activity has been associated with common vascular diseases such as atherosclerotic plaque formation, abdominal aortic aneurysms, and critical limb ischemia. For this reason, MMPs have become an important focus for basic science studies and clinical investigations by vascular biology researchers. This article reviews the recent literature, summarizing our current understanding of the role of MMPs in the pathogenesis of various peripheral vascular disease states. In addition, the importance of MMPs in the future diagnosis and treatment of peripheral vascular disease is discussed. PMID- 17398402 TI - Fiduciary economization: your wealth or your health. PMID- 17398404 TI - Regarding "Reduced recanalization rates of the great saphenous vein after endovenous laser treatment with increased energy dosing: definition of a threshold for the endovenous fluence equivalent". PMID- 17398405 TI - Vocal cord paralysis after aortic arch surgery with stent-graft placement, a contemporary method of arch surgery. PMID- 17398407 TI - Abstracts from the 15th Association of European Psychiatrists Congress. March 17 21, 2007. Madrid, Spain. PMID- 17399619 TI - Suprascapular nerve palsy after the Latarjet procedure. PMID- 17399620 TI - Delayed radial head dislocation associated with malunion of radial shaft fracture: a case report. PMID- 17399621 TI - Rehabilitation of two-part fractures of the neck of the humerus (two-year follow up). AB - The 2-year results of a randomized, prospective, controlled trial of minimally displaced proximal humeral fractures treated either by immediate physiotherapy (group A) or after 3 weeks of immobilization (group B) are reported. At 1 year shoulder disability, as measured with the Croft shoulder disability questionnaire, was found in 42.8% of patients in group A and 72.5% in group B (P < .01). By 2 years, shoulder disability in group A remained unchanged (43.2%) but had reduced in group B (59.5%). This difference was not statistically significant. Immediate physiotherapy after a minimally displaced proximal humeral fracture results in faster recovery, with maximal functional benefit being achieved at 1 year. Delayed rehabilitation by 3 weeks of shoulder immobilization produces a slower recovery, which continues for at least 2 years after the time of injury. PMID- 17399622 TI - Intraoperative localization of calcification in the supraspinatus via a percutaneous marking suture after preoperative ultrasound. AB - In a prospective study, we evaluated the efficiency of a technique for localizing calcification via a marking suture. In 27 patients, we measured the distance between the center of the calcification and the long head of the biceps preoperatively. Intraoperatively, we determined the distance detected by ultrasound on the articular side of the cuff under arthroscopic visualization. In this position, a spinal needle was inserted through the supraspinatus tendon. A PDS suture was passed through the spinal needle into the joint. The spinal needle was then removed. The entrance point of the PDS suture into the supraspinatus tendon was localized in the subacromial space after resection of bursal tissue. We perforated the cuff with a spinal needle in this area to definitively detect the calcification. The calcification was pressed out of the cuff with a probe. With this technique, calcification could be removed arthroscopically in all patients. This operative technique led to a reduced operative time, with a mean of 19 minutes. PMID- 17399623 TI - Rotator cuff repair: an analysis of utility scores and cost-effectiveness. AB - More than 75,000 rotator cuff repairs are performed annually, yet cost effectiveness data are unavailable. This study examines the cost utility of rotator cuff repair by relating surgical costs to increase in quality-adjusted life-years (QALYs). Eighty-seven patients were followed up prospectively for 1 year, during which cost and quality of life data were collected. Patient-based utility measures of quality of life (European Quality-of-Life measure [EuroQoL] and Health Utility Index [HUI]) were administered. Changes in these measures generated net QALYs. Finally, life expectancies were applied to generate a cost effectiveness ratio, and subsequent 1-way sensitivity analyses varied costs, QALYs, and discount rates to determine which factors drive cost-effectiveness. Total costs averaged $10,605.20. Significant improvements were noted in health related quality of life postoperatively. The estimated mean lifetime gain in QALYs from surgery was 0.81 by use of the HUI and 3.43 by use of the EuroQoL. This yielded cost-effectiveness ratios of $13,092.84/QALY by use of the HUI and $3,091.90/QALY by use of the EuroQoL. The cost-effectiveness of rotator cuff repair compares favorably with other common interventions in health care and reaches commonly accepted benchmarks for cost-effectiveness. PMID- 17399624 TI - Anterior shoulder instability with humeral avulsion of the glenohumeral ligament lesion. AB - Humeral avulsion of the glenohumeral ligament (HAGL) is a rare lesion. The purpose of this study was to analyze the clinical manifestations of HAGL lesions in patients who underwent operative treatment for anterior shoulder instability. Six patients with HAGL lesions were studied. Four patients had an HAGL lesion associated with a Bankart lesion, and two had an isolated HAGL lesion. The range of motion at final follow-up showed a loss of 1 degree in forward flexion and of 15 degrees in external rotation. During an operation to treat anterior shoulder instability, a thorough examination for not only Bankart lesions but also other associated lesions, including an HAGL lesion, should be considered to lower the risk of redislocation. In repairing an HAGL lesion, the surgeon should keep in mind the possibility of a postoperative loss of external rotation and follow an active rehabilitation protocol to obtain successful results. PMID- 17399625 TI - The effect of suture fixation of type I coronoid fractures on the kinematics and stability of the elbow with and without medial collateral ligament repair. AB - The objective of this study was to determine the effect of suture repair of type 1 coronoid fractures on elbow kinematics in ligamentously intact and medial collateral ligament (MCL)-deficient elbows. Cadaveric testing was performed in stable and MCL-deficient elbows with radial head arthroplasty and with the coronoid intact, with the coronoid fractured, and after suture repair. Ulna versus humerus angulation was measured during active motion. Varus and valgus motion pathways were measured during passive gravity-loaded flexion. With intact ligaments, there was a small increase in valgus angulation after a type 1 fracture that was not corrected with suture fixation. With MCL deficiency, there was no change in kinematics regardless of coronoid status. Type 1 coronoid fractures cause only small changes in elbow kinematics that are not corrected with suture repair. MCL repair, rather than type 1 coronoid fixation, should be considered if the elbow remains unstable after radial head repair or replacement and lateral ligament repair. PMID- 17399626 TI - Biomechanical evaluation of locked plating in proximal humeral fractures. AB - The ideal method of fixation in proximal humeral fractures is not well established. Locked plating is a potential solution, but to date, there has been little support shown in the literature. We compared the biomechanical performance of a locked plate with that of an intramedullary humeral nail in a cadaveric model. Paired fresh-frozen cadaveric humeri (N = 5) were instrumented with either a locked plate or an intramedullary nail after creation of a 3-part osteotomy. Construct stiffness, cyclic loading behavior, and load to failure were evaluated. The locked plate was more stiff in valgus loading than the nail (P = .003). All other loading vectors showed no statistical differences in stiffness between the two constructs. The load to failure was 828 N for the plate and 901 N for the nail. The nail showed interfragmentary motion (degrees), or toggle, at zero load, whereas the plate did not. The locked plate, being stable at zero load, may prove useful in earlier pain-free rehabilitation of proximal humeral fractures. PMID- 17399627 TI - Atypical atrioventricular nodal reentrant tachycardia with eccentric coronary sinus activation: electrophysiological characteristics and essential effects of left-sided ablation inside the coronary sinus. AB - BACKGROUND: The precise electrophysiological characteristics and essential effects of left-sided ablation in atrioventricular nodal reentrant tachycardia (AVNRT) with eccentric coronary sinus (CS) activation (ECSA) have not been described. OBJECTIVE: The purpose of this study was to elucidate the tachycardia characteristics and essential effects of left-sided ablation in AVNRT with ECSA. METHODS: Electrophysiological and ablation data were reviewed in 340 patients with all forms of AVNRT. RESULTS: Among 360 AVNRTs in the 340 patients, there were 23 atypical AVNRTs (6%; 12 slow-slow and 11 fast-slow) in 18 (5%) patients who exhibited ECSA with the earliest retrograde atrial activation 11 +/- 5 mm inside the CS. The patients with ECSA during the tachycardia were significantly younger than those without (38 +/- 18 vs. 51 +/- 18 years; P<.01). The presence of upper (UCP) and lower common pathways (LCP) was suggested in three (17%) and 18 (100%) patients, respectively. An ablation exclusively targeting the earliest retrograde atrial activation inside the CS eliminated the tachycardias with the elimination (n = 12) or modification of the left-sided slow pathway (SP) conduction (n = 6) without any complications. The entire reentrant circuit was considered to reside on the left side in two patients (11%) because the bidirectional SP conduction was simultaneously eliminated after the ablation inside the CS. CONCLUSIONS: Atypical AVNRT with ECSA involved the left-sided SP as a retrograde limb, and the reentrant circuit was more frequently associated with evidence that suggested a UCP and LCP. Ablation exclusively targeting the earliest retrograde atrial activation inside the CS was highly effective in this entity. PMID- 17399628 TI - Atypical atrioventricular nodal reentry with eccentric atrial activation. Is the right target on the left? PMID- 17399629 TI - Multiple focal atrial tachycardias in a healthy adult population: characterization and description of successful radiofrequency ablation. AB - BACKGROUND: The electrophysiologic (EP) characteristics and ablation outcomes of focal atrial tachycardia (AT) have been extensively characterized over recent years. However, there are limited published data describing patients who at EP study have more than one sustained focal tachycardia. OBJECTIVE: To characterize the demographics, tachycardia data, and outcome of patients with successful ablation of more than one focal AT. METHODS: A retrospective review of our supraventricular tachycardia database from 2000 to 2006 identified 258 patients who had undergone radiofrequency ablation of focal AT. Ten patients were identified who had more than one sustained focal tachycardia at EP study, including seven patients with two ATs and three patients with three ATs. The patients were all women with a mean age of 54.2 +/- 10.3 years. AT locations included the crista terminalis, coronary sinus ostium, tricuspid and mitral annuli, and pulmonary vein ostium. Successful ablation was performed for 22 (95.7%) of 23 tachycardias. RESULTS: During long-term follow-up of 32.7 +/- 22.2 months, there were no recurrences of AT in those patients with successful ablation. No patients developed new AT foci, and only one developed late AF. CONCLUSIONS: We have described a series of patients with multiple focal ATs with typical anatomic distribution. These patients do not have significant cardiac or respiratory disease, and in this series, all were women. This report demonstrates that up to three focal ATs can be successfully ablated at a single procedure without recurrence or development of other atrial arrhythmias in long-term follow up. PMID- 17399630 TI - Increased incidence of subacute lead perforation noted with one implantable cardioverter-defibrillator. AB - BACKGROUND: The rapid evolution of implantable cardioverter-defibrillator (ICD) leads has resulted in thinner active fixation leads. While these advances have made the leads more versatile, new configurations may be associated with unforeseen complications. OBJECTIVE: The purpose of this study was to determine the incidence of perforation and dislodgement of defibrillator leads in a single center in the year 2005. METHODS: All patients who underwent percutaneous ICD implantation at the Massachusetts General Hospital using an endocardial right ventricular lead were included in this study. The specific leads analyzed were the Riata (1580/1581 and 1590/1591, St. Jude Medical, St Paul, Minnesota, USA;) and Sprint Fidelis (6949-65, Medtronic, Minneapolis, Minnesota, USA.). Information was collected retrospectively. RESULTS: A total of 130 Riata leads and 111 Sprint Fidelis leads were implanted at the Massachusetts General Hospital during this time period. A total of five lead perforations occurred in patients implanted with the Riata lead as compared with none with the Sprint Fidelis lead (3.8% vs. 0%, respectively; P <.05). Two of the five patients with perforation required pericardiocentesis for tamponade. Clinical symptoms of perforation developed 1-10 days after implant. Moreover, there were five additional lead revisions in the Riata group, which were likely due to dislodgement and/or microperforation, as compared with none in the Sprint Fidelis group (7.7% vs. 0%, respectively; P <.005). CONCLUSIONS: In 2005, at one institution, there were significantly more cardiac perforations and lead revisions with the Riata lead as compared with the Sprint Fidelis right ventricular defibrillator lead. Further data are required to determine whether certain lead characteristics are responsible for this observation. PMID- 17399631 TI - The importance of retrospective analyses of adverse outcomes. PMID- 17399632 TI - Localizing circuits of atrial macroreentry using electrocardiographic planes of coherent atrial activation. AB - BACKGROUND: The complexity of ablation for atrial macroreentry atrial flutter (AFL) varies significantly depending on the circuit location. Presently, surface electrocardiogram (ECG) analysis poorly separates left from right atypical AFL and from some cases of typical AFL, which delays diagnosis until invasive study. OBJECTIVE: The purpose of this study was to differentiate and localize the intra atrial circuits of left atypical AFL, right atypical, and typical AFL using quantitative ECG analysis. METHODS: We studied 66 patients (54 men, age 59 +/- 14 years) with typical (n = 35), reverse typical (n = 4), and atypical (n = 27) AFL. For each, we generated filtered atrial waveforms from ECG leads V5 (X-axis), aVF (Y-axis), and V1 (Z-axis) by correlating a 120-ms F-wave sample to successive ECG regions. Atrial spatial loops were plotted for three orthogonal planes (frontal, XY = V5/aVF; sagittal, YZ = aVF/V1; axial, XZ = V5/V1) and then cross-correlated to measure spatial regularity (i.e., coherence; range -1 to 1). RESULTS: Mean coherence was greatest in the XY plane (P <10(-3) vs. XZ or YZ). Atypical AFL showed lower coherence than typical AFL in the XY (P <10(-3)), YZ (P <10(-6)), and XZ (P <10(-5)) planes. Atypical left AFL could be separated from atypical right AFL by lower XY coherence (P = .02); for this plane, coherence <0.69 detected atypical left AFL with 84% specificity and 75% sensitivity. F-wave amplitude alone did not separate typical, atypical right, or atypical left AFL (P = NS). CONCLUSIONS: Atypical AFL shows lower spatial coherence than typical AFL, particularly in the sagittal and axial planes. Coherence in the Cartesian frontal plane separated left and right atypical AFL. Such analyses may be used to plan ablation strategy from the bedside. PMID- 17399633 TI - Spatiotemporal loops: a new three-dimensional mapping system? PMID- 17399634 TI - Transseptal endocardial left ventricular pacing: an alternative technique for coronary sinus lead placement in cardiac resynchronization therapy. AB - BACKGROUND: Coronary sinus (CS) lead placement for transvenous left ventricular (LV) pacing in cardiac resynchronization therapy (CRT) has a failure rate at implant and short-term follow-up between 10% and 15%. OBJECTIVE: The purpose of this study was to assess the feasibility of transseptal endocardial LV pacing in patients in whom transvenous CS lead placement had failed. METHODS: An atrial transseptal LV lead placement was attempted in 10 patients (six females, age 69.4 +/- 9.6 years), in whom CS lead placement for CRT had failed. After transseptal puncture and septal dilatation from the femoral route, the left atrium was cannulated with a combination of catheters and guide wires from the left or right subclavian vein. After advancement of this guide catheter into the LV, a standard bipolar screw-in lead could be implanted in the posterolateral wall. All patients were maintained on anticoagulant therapy with warfarin after implant. RESULTS: An LV lead could be successfully implanted in nine of the 10 patients. The stimulation threshold was 0.78 +/- 0.24 V, and the R-wave amplitude was 14.2 +/- 9.7 mV. At 2 months' follow-up, the stimulation threshold was 1.48 +/- 0.35 V with a 0.064 +/- 0.027 ms pulse width. There was no phrenic nerve stimulation observed in any of the patients. There were no thromboembolic complications at follow-up. CONCLUSIONS: LV transseptal endocardial lead implantation from the pectoral area is a feasible approach in patients with a failed CS approach and in whom epicardial surgical lead placement is not an option. Longer follow-up is warranted to determine the risk of thromboembolic complications. PMID- 17399635 TI - Effect of perioperative corticosteroid use on the incidence of postcardiothoracic surgery atrial fibrillation and length of stay. AB - BACKGROUND: While the mechanism underlying the development of post-cardiothoracic surgery atrial fibrillation has not been fully elucidated, it has been suggested that inflammation may play a causative role. Corticosteroids have been traditionally used to reduce inflammation, and when administered perioperatively, they may decrease the incidence of atrial fibrillation. OBJECTIVE: The purpose of this study was to investigate the effects of corticosteroid administration on the incidence of post-cardiothoracic surgery atrial fibrillation and resultant hospital length of stay. METHODS: A systematic literature search of MEDLINE, EMBASE, Web of Science, and the Cochrane Database of Systematic Reviews through July 2006 was conducted using specific search terms. A review of cardiology abstracts and a manual review of references were also performed. Studies that met the following criteria were included: randomized, controlled trials comparing patients receiving perioperative corticosteroids or placebo and reporting data on post-cardiothoracic surgery atrial fibrillation. A random-effects model was used. RESULTS: Nine of the 1396 citations initially identified, representing 990 patients, met the inclusion criteria. Upon meta-analysis, corticosteroids significantly lowered patients' odds of developing post-cardiothoracic surgery atrial fibrillation by 45% (odds ratio [OR] 0.55; 95% confidence interval [CI] 0.39-0.78) and reduced hospital length of stay by approximately 1.6 days (weighted mean difference -1.59; 95% CI -2.96 to -0.21). Reductions in the incidence of post-cardiothoracic surgery atrial fibrillation appeared greatest in patients receiving intermediate doses of corticosteroid (50-210 mg dexamethasone equivalent), while both lower (up to 8 mg) and higher (236-2850 mg) dosing resulted in blunted effects. CONCLUSION: Corticosteroids appear to reduce the incidence of post-cardiothoracic surgery atrial fibrillation and shorten hospital length of stay in randomized controlled trials. PMID- 17399636 TI - Mapping a novel locus for familial atrial fibrillation on chromosome 10p11-q21. AB - BACKGROUND: Atrial Fibrillation (AF), the most common cardiac arrhythmia, is a significant public health problem in the United States, affecting approximately 2.2 million Americans. Recently, several chromosomal loci and genes have been found to be associated with familial AF. However, in most other AF cases, the genetic basis is still poorly understood. OBJECTIVE: The purpose of this study was to investigate the molecular basis of familial AF in a Dutch kindred group. METHODS: We analyzed a four-generation Dutch family in which AF segregated as an autosomal dominant trait. After the exclusion of linkage to 10q22-24, 6q14-16, 5p13, KCNQ1, KCNE2, KCNJ2 and some ion-channel-associated candidate genes, a genome-wide linkage scan using 398 microsatellite markers was performed. RESULTS: Two-point logarithms of odds (LOD) scores >1 at recombination fraction [theta] = 0.00 and a haplotype segregating with the disorder were demonstrated only across regions of chromosome 10. Subsequent fine mapping gave a maximum two-point LOD score of 4.1982 at D10S568 at [theta] = 0.00. Distinct recombination in several individuals narrowed the shared region among all affected individuals to 16.4 cM on the Genethon map (flanking markers: D10S578 and D10S1652), which corresponds to chromosome 10p11-q21. Thirteen candidate genes residing in this region, which could be associated with AF, were screened. No mutation has been found in their coding regions including the intron splice regions. CONCLUSION: We identify a novel locus for AF on chromosome 10p11-q21, which provides further evidence of genetic heterogeneity in this arrhythmia. PMID- 17399637 TI - Confronting the genetic complexity of atrial fibrillation: this too shall pass. PMID- 17399638 TI - Direct measurements of membrane time constant during defibrillation strength shocks. AB - BACKGROUND: Defibrillation shocks impose significant energy demand on implantable cardioverter-defibrillators (ICDs). Several modeling studies have been devoted to optimizing shock parameters, and a large number of these studies treat the heart as a simplified lumped network. The time constant of membrane polarization (tau(m)) is a key variable for such modeling efforts. OBJECTIVE: The purpose of this study was to perform direct measurements of transmembrane potential (V(m)) during defibrillation strength shocks and estimate tau(m) of membrane polarization. METHODS: A portion of the left ventricular epicardial surface of Langendorff-perfused rabbit hearts was stimulated using uniform electric fields produced by two parallel line electrodes. The V(m)s were recorded from di-4 ANEPPS-stained hearts using a multisite optical mapping system. The hearts were paced with 20 S1 pulses from the apex, and shocks (S2: 5, 10, 20 V/cm) were applied via the line electrodes during the action potential of the 20th S1 at two different coupling intervals (S1S2: 120 and 180 ms). Residual responses were obtained by subtracting responses to the 19th S1 from the responses to the 20th S1S2 pair and used for time-constant analysis by fitting a monoexponential function. RESULTS: tau(m) exhibited a large variation and ranged from approximately 1 to 30 ms. Furthermore, tau(m) varied with electric field strength, S1S2 interval, position of the tissue from stimulating electrodes, and polarity of the response. To a large extent, the effects of all these factors were captured in a single parameter-the change in transmembrane voltage (DeltaV(m)) in response to the applied field (E). tau(m) showed a monotonically decreasing trend with DeltaV(m) for all Es and S1S2s. CONCLUSION: Time constant of membrane polarization varies significantly during defibrillation strength shocks and shows a strong dependence on DeltaV(m). PMID- 17399639 TI - Kir2.3 isoform confers pH sensitivity to heteromeric Kir2.1/Kir2.3 channels in HEK293 cells. AB - BACKGROUND: Data on pH regulation of the cardiac potassium current I(K1) suggest species-dependent differences in the molecular composition of the underlying Kir2 channel proteins. OBJECTIVE: The purpose of this study was to test the hypothesis that the presence of the Kir2.3 isoform in heterotetrameric channels modifies channel sensitivity to pH. METHODS: Voltage clamp was performed on HEK293 cells stably expressing guinea pig Kir2.1 and/or Kir2.3 isoforms and on sheep cardiac ventricular myocytes at varying extracellular pH (pH(o)) and in the presence of CO(2) to determine the sensitivity of macroscopic currents to pH. Single-channel activity was recorded from the HEK293 stables to determine the mechanisms of the changes in whole cell current. RESULTS: Biophysical characteristics of whole-cell and single-channel currents in Kir2.1/Kir2.3 double stables displayed properties attributable to isoform heteromerization. Whole-cell Kir2.1/Kir2.3 currents rectified in a manner reminiscent of Kir2.1 but were significantly inhibited by extracellular acidification in the physiologic range (pK(a) approximately 7.4). Whole-cell currents were more sensitive to a combined extracellular and intracellular acidification produced by CO(2). At pH(o) = 6.0, unitary conductances of heteromeric channels were reduced. Ovine cardiac ventricular cell I(K1) was pH(o) and CO(2) sensitive, consistent with the expression of Kir2.1 and Kir2.3 in this species. CONCLUSION: Kir2.1 and Kir2.3 isoforms form heteromeric channels in HEK293. The presence of Kir2.3 subunit(s) in heteromeric channels confers pH sensitivity to the channels. The single and double stable cells presented in this study are useful models for studying physiologic regulation of heteromeric Kir2 channels in mammalian cells. PMID- 17399640 TI - Physiologic function of I(K1) requires a combination of Kir2 isoforms. PMID- 17399641 TI - Ionic mechanisms underlying region-specific remodeling of rabbit atrial action potentials caused by intermittent burst stimulation. AB - BACKGROUND: Pulmonary veins (PVs) and the coronary sinus (CS) play pivotal roles in triggering some episodes of atrial fibrillation. In isolated rabbit right or left atrial preparations, a 3-hour intermittent burst pacing protocol shortens action potential duration (APD) in CS and PV, but not in sinus node (SN) and left Bachmann bundle (BB) regions. OBJECTIVE: The purpose of this study was to use patch clamp techniques to study the rapidly inactivating (I(to)) and sustained (I(sus)) K(+) currents as well as Ca(2+) currents (I(Ca)) in cells dispersed from intermittent burst pacing and sham PV, BB, CS, and SN regions to determine whether changes in these currents contributed to APD shortening. METHODS: Real time polymerase chain reaction was performed for transient outward K(+) and Ca(2+) channel subunit mRNAs to determine if intermittent burst pacing affected expression levels. RESULTS: I(to) densities were unaffected by intermittent burst pacing in PV and Bachmann bundle cells. mRNA levels of K(V)4.3, K(V)4.2, K(V)1.4, and KChIP2 subunits of I(to) in both regions were stable. In CS cells, I(to) densities in intermittent burst pacing were greater than in sham (P <.05), but there were no parallel mRNA changes. I(Ca) density of PV cells was reduced from 14.27 +/- 2.08 pA/pF (at -5 mV) in sham to 7.52 +/- 1.65 pA/pF in intermittent burst pacing PV cells (P <.05) due to a significant shift in voltage dependence of activation. These results were seen in the absence of mRNA changes in alpha(1C) and alpha(1D) Ca(2+) channel subunits. In contrast, intermittent burst pacing had no effect on Ca(2+) current densities and kinetics of CS cells, but decreased alpha(1)C and alpha(1)D mRNA levels. CONCLUSION: There is region specific remodeling of I(to) and I(Ca) by intermittent burst pacing protocols in rabbit atrium. Increased I(to) in CS cells could account for the APD shortening observed with intermittent burst pacing, whereas an intermittent burst pacing induced shift in voltage dependence of activation may contribute to APD shortening in PV cells. PMID- 17399642 TI - Flecainide for recurrent malignant ventricular arrhythmias in two siblings with Andersen-Tawil syndrome. PMID- 17399643 TI - Electrophysiologic characteristics of an Andersen syndrome patient with KCNJ2 mutation. AB - We report the first case of a patient with Andersen syndrome in whom electrophysiologic study was performed. The patient was a 19-year-old woman with familial periodic paralysis, abnormal QT-U complex, and nonsustained ventricular tachycardia. Mutation analysis revealed a missense mutation in KCNJ2, a component of Kir2.1. Monophasic action potential recordings showed a delayed afterdepolarization (DAD)-like hump in the left ventricle. Initiation of epinephrine-induced premature ventricular contractions always coincided with both the exaggerated DAD-like hump and the U wave. These findings suggest that reduced Kir2.1 current contributes to the development of DAD and ventricular arrhythmias in Andersen syndrome. PMID- 17399644 TI - Novel SCN5A mutation (Q55X) associated with age-dependent expression of Brugada syndrome presenting as neurally mediated syncope. AB - BACKGROUND: An association between Brugada syndrome and neurally mediated syncope has been described. Although mutations in SCN5A have been identified in Brugada syndrome, the genetic link between Brugada syndrome and neurally mediated syncope has not been determined. OBJECTIVES: The purpose of the study was to clinically and genetically characterize a man with recurrent syncope that originally was diagnosed as neurally mediated syncope at age 8 years but subsequently manifested as Brugada syndrome at age 17 years. METHODS: The proband underwent clinical examination, which included head-up tilt test, sodium channel provocation test, and electrophysiologic study. Genetic screening of SCN5A was performed for the proband and his family members. The biophysical properties of a mutant SCN5A channel in a heterologous expression system were studied using whole-cell, patch clamp technique. RESULTS: The proband showed positive head-up tilt test, coved type ST elevation recorded from the third intercostal space, and positive pilsicainide provocation test. Ventricular fibrillation was inducible at programmed electrical stimulation, consistent with characteristics of both Brugada syndrome and neurally mediated syncope. A novel nonsense SCN5A mutation (Q55X) was identified in the proband, his mother, and his asymptomatic brother. The heterologously expressed mutant channel was nonfunctional. CONCLUSION: We genetically determined an SCN5A mutation in a patient showing the combined phenotype of neurally mediated syncope and Brugada syndrome. Neurally mediated syncope and Brugada syndrome may share, at least in part, a common pathophysiologic mechanism. PMID- 17399645 TI - Protagonist: routine screening of all athletes prior to participation in competitive sports should be mandatory to prevent sudden cardiac death. PMID- 17399646 TI - Antagonist: routine screening of all athletes prior to participation in competitive sports should be mandatory to prevent sudden cardiac death. PMID- 17399647 TI - Mechanically induced ventricular fibrillation (commotio cordis). PMID- 17399648 TI - Expeditious evaluation of paroxysmal supraventricular tachycardia: what is the mechanism? PMID- 17399649 TI - Cycle length alternans during ventricular pacing. PMID- 17399650 TI - Pulmonary vein-related maneuvers: part I. PMID- 17399651 TI - Atrioventricular nodal reentrant tachycardia with a varying H-H interval, a fixed atrial cycle length and 2:1 A-V relation. PMID- 17399652 TI - Inter-association task force recommendations on emergency preparedness and management of sudden cardiac arrest in high school and college athletic programs: a consensus statement. AB - OBJECTIVE: To assist high school and college athletic programs prepare for and respond to a sudden cardiac arrest (SCA). This consensus statement summarizes our current understanding of SCA in young athletes, defines the necessary elements for emergency preparedness, and establishes uniform treatment protocols for the management of SCA. BACKGROUND: Sudden cardiac arrest is the leading cause of death in young athletes. The increasing presence of and timely access to automated external defibrillators (AEDs) at sporting events provides a means of early defibrillation and the potential for effective secondary prevention of sudden cardiac death. An Inter-Association Task Force was sponsored by the National Athletic Trainers' Association to develop consensus recommendations on emergency preparedness and management of SCA in athletes. RECOMMENDATIONS: Comprehensive emergency planning is needed for high school and college athletic programs to ensure an efficient and structured response to SCA. Essential elements of an emergency action plan include establishing an effective communication system, training of anticipated responders in cardiopulmonary resuscitation and AED use, access to an AED for early defibrillation, acquisition of necessary emergency equipment, coordination and integration of onsite responder and AED programs with the local emergency medical services system, and practice and review of the response plan. Prompt recognition of SCA, early activation of the emergency medical services system, the presence of a trained rescuer to initiate cardiopulmonary resuscitation, and access to early defibrillation are critical in the management of SCA. In any collapsed and unresponsive athlete, SCA should be suspected and an AED applied as soon as possible for rhythm analysis and defibrillation if indicated. PMID- 17399653 TI - The tuberous breast revisited. AB - BACKGROUND: The tuberous breast presents a problem for which many surgical solutions have been described. Current teaching describes how the tuberous breast deformity is the result of skin shortage as well as herniation of breast tissue through the nipple-areola complex. However, through careful clinical observation we now believe that the only abnormality present is herniation of breast tissue through the nipple-areola complex. METHODS: Using this principle, we have refined a one-stage surgical procedure that can be used to correct any type of tuberous breast deformity. Since 2001 we have performed our technique on a series of 13 tuberous breasts of widely varying appearances in eight patients (age 17-24 years) with a follow up varying between 3 and 56 months. Our new understanding of the tuberous breast deformity has also made it possible to develop an objective, reproducible method for defining the tuberous breast based on the degree of areola herniation. RESULTS: All patients reported high levels of satisfaction with the procedure. Assessment of the results by an independent panel of attending surgeons showed all results to be good/excellent. Moreover, the results have improved with time and no revisions have been needed. Our method of defining the tuberous breast (based on the ratio of areola herniation:areola diameter) enabled us to identify a cut-off to decide (objectively) when a breast was tuberous. This allowed us to anticipate when an areola reduction/tightening procedure would be necessary to avoid a 'double-bubble' deformity. CONCLUSION: We propose a one-stage surgical procedure which is applicable to all degrees of tuberous breast deformity. The results appear to confirm our theory that the only abnormality present in the tuberous breast is herniation of breast tissue through the nipple-areola complex. In patients with small breasts and a tuberous deformity, correction of the herniation changes the tuberous breast into a simple hypoplastic breast. The volume deficit can then be corrected by augmentation (if desired by the patient). In patients with sufficient breast volume, correction of the herniation alone will correct the deformity. PMID- 17399654 TI - Breastfeeding after vertical reduction mammaplasty using a superior pedicle. AB - Between January 1996 and December 2002, 189 women underwent bilateral superior pedicle breast reduction according to the Lejour technique, at Brugmann University Hospital (Brussels, Belgium). We conducted a retrospective study on the 18 women who gave birth since the operation. Our aim was to find out how many of them breastfed, the reasons for not breastfeeding from those who did not, and to look for parameters that might have interfered with breastfeeding success. The latter was defined as the ability to breastfeed for a minimum of 2 weeks, with or without nutritional supplementation. Eight patients out of 18 successfully breastfed for an average duration of 1.9+/-1.3 months, and ceased to do so in most cases for reasons other than spontaneous cessation of lactation. Another eight women made no attempt at breastfeeding for various reasons. According to data obtained from a report of the Belgian French-speaking Community, our series of patients made proportionally fewer breastfeeding attempts than women from a culturally similar context (74%). However, they exclusively breastfed for a median duration of 3 months, superior to the 8-9 weeks of the women included in the government report. Two patients failed at their breastfeeding attempts. We were able to demonstrate the role of neither anthropomorphic, operation-related, neuro-sensitive nor psychological factors in those failures. Our results, when confronted to those of similar studies, show that the Lejour technique allows breastfeeding in proportions similar to those of any other pedicled technique. PMID- 17399655 TI - Refinements of the LeJour vertical mammaplasty skin pattern for skin-sparing mastectomy and immediate breast reconstruction. AB - BACKGROUND: Skin-sparing mastectomy (SSM) is a well-established technique for immediate breast reconstruction (IBR). When used for large and/or ptotic breasts, traditional SSM patterns produce long skin flaps prone to necrosis or 'T' junction breakdown. The authors have previously demonstrated the applicability of the LeJour-type vertical mammaplasty skin pattern to this group of patients. With further experience, indications for this procedure have been widened and the technique refined. RESULTS: Over five years, 26 immediate breast reconstructions were carried out in 19 patients using this technique: three expandable implants, seven LDs, three pedicled TRAMs, five free TRAMs, seven DIEPs and one SIEA flap. Fourteen patients (74%) had simultaneous contralateral balancing LeJour breast reductions or mastopexies. The remaining five patients had bilateral mastectomies and reconstructions using the vertical mammaplasty skin pattern for both breasts. All flaps were successful, but there were three cases of minor skin flap necrosis, three of delayed wound healing and two instances of significant post operative bleeding. Cosmesis was suboptimal in the prosthetic reconstruction group, necessitating revisional surgery. DISCUSSION AND CONCLUSIONS: The vertical mammaplasty skin pattern was successfully used with a wide range of reconstructions. However, to avoid suboptimal cosmetic results and minimise wound healing problems this technique is not recommended in heavy smokers, very obese patients, those undergoing prosthetic reconstructions or neoadjuvant chemotherapy. The skin resection pattern should also be conservative. The LeJour type vertical mammaplasty pattern is a viable alternative technique for SSM in selected patients, especially those requiring contralateral balancing surgery and undergoing autologous tissue reconstruction. PMID- 17399656 TI - Soft cohesive silicone gel breast prostheses: a comparative prospective study of aesthetic results versus lower cohesivity silicone gel prostheses. AB - The flexibility of lower cohesivity silicone prostheses is the main reason for wrinkling, rippling and evidence of implant edges. The soft cohesive silicone implants promise to minimize such effects with minimal softness reduction. Forty consecutive patients received soft cohesive prostheses (INAMED Style 110 ST) and were studied prospectively. A historical group, made up by the 40 consecutive patients who received lower cohesivity silicone implants (INAMED Style 110) in the immediately preceding months, was used as a control. Wrinkling, prosthetic edge perceptibility and capsular contracture degree were assessed six months after surgery. The tissue coverage thickness was measured using ultrasonography. The patients were then asked to evaluate the breast softness by means of an anonymous questionnaire, where they also expressed their overall satisfaction by means of the five-steps linear analogical scales. The wrinkling prevalence was 9.2% in the soft cohesive group vs. 55% in the lower cohesivity one (p<0.01). The edge perceptibility was 14% in the soft cohesive group vs. 22% in the lower cohesivity one (no statistical significance). The coverage tissue thickness was not found to be significantly related to the wrinkling prevalence or to the edge perceptibility. The capsular contracture rate was almost identical in the two groups (Baker II: 2.6% vs. 2.7%, no Baker III or IV). A higher stiffness was noted in the soft cohesive group (average score: 4.2 vs. 4.4 in the control group, p<0.05), but the overall satisfaction degree was higher for soft cohesive implants (average score: 4.5 vs. 3.8, p<0.01). The soft cohesive prostheses offered better overall results than the lower cohesivity silicone prostheses, even if a longer term follow-up should be advised. The soft cohesive prostheses showed a higher firmness, but this seemed not to have any influence on the overall satisfaction degree. PMID- 17399657 TI - Erysipelas-like inflammation following breast surgery. AB - Impaired lymph drainage is an inevitable consequence of any form of surgery that disrupts lymphatics, resulting in a degree of lymphoedema that may vary from subtle to dramatic and although classically involving an entire limb, may be more localised, confined to only a small area such as a skin flap. Infection is a well recognised complication of lymphoedema. However, not all inflammatory episodes occurring in the setting of lymphatic dysfunction can be clearly attributed to infection as this article demonstrates. Five patients presented over a 5-year period with distinctive erysipelas-like inflammation affecting the breast which occurred several weeks following reduction mammaplasty in four patients and breast reconstruction in one patient. No clinical response was obtained with standard antibiotics. This inflammatory problem may represent a previously unreported complication of breast surgery with an incidence of 4% following reduction mammaplasty. Recent research supports the notion that this type of episode is most likely to be due to a non-infective inflammatory process related to lymphatic dysfunction induced by surgery. PMID- 17399658 TI - Early experience of immediate reconstruction using autologous free dermal fat graft after breast conservational surgery. AB - Breast conservational therapy (BCT) has become a standard strategy for breast cancer, and ensures that local control with acceptable cosmetic results [Fisher B, Anderson S, Redmond CK, et al. Reanalysis and results after 12 years of follow up in a randomized clinical trial comparing total mastectomy with lumpectomy with or without irradiation in the treatment of breast cancer. N Engl J Med 1995;333:1456-61] and immediate reconstruction after BCT has become increasingly popular even for early-stage breast cancer [Berrino P, Campora E, Santi P. Post quadrantectomy breast deformities: classification and techniques of surgical correction. Plast Reconstr Surg 1987;79(4):567-72; Cooperman AM, Dinner M. The rhomboid flap and partial mastectomy. Surg Clin North Am 1978;58:869-73]. The breasts of Japanese women are sometimes too small to maintain symmetry, even after partial resection, and an insufficient resection margin may increase local recurrence if too much attention is paid to cosmesis. Use of an autologous free dermal fat graft (FDFG) for defect reconstruction has proven popular for some areas of the body [Lexer E. Free transplantation. Ann Surg 1914;60:166-94; Peer LA. The neglected free fat graft. Plast Reconstr Surg 1956;18:233-50]. Immediate reconstruction of the surgical defect was performed in seven Japanese women using autologous free dermal fat graft (FDFG) from the lower abdomen after breast conserving surgeries for six malignant lesions and two benign masses located in the medial or central area of the breasts. The mean amount of resected tissue was 56.4 g (range, 28-108 g), while mean FDFG weight was 78.3g (range, 35-148 g). We obtained symmetry in the size of the whole breast, the position and level of the nipple, and shape. Autologous FDFG was useful for reconstruction, with a good cosmetic effect. This technique achieves better cosmetic results than the transposition of residual breast tissue, is more convenient than muscle flap grafting, and safer than implantation of foreign materials. This report documents our early experiences with seven patients who underwent BCT and immediate breast reconstruction using autologous FDFG. PMID- 17399659 TI - Prospective analysis of psychosocial outcomes after breast reconstruction: cross cultural comparisons of 1-year postoperative results. AB - Breast reconstruction has been shown to improve quality of life in women following mastectomy for breast cancer. To date, there have been no published prospective reports looking at the effect nationality has on patient quality of life following breast reconstruction. Women from the USA, Sweden and Canada were recruited prior to reconstruction and followed prospectively for 1 year postoperatively. Thirteen centres with 24 plastic surgeons were involved. Preoperatively and 1 year postoperatively, women completed the Short Form-36 questionnaire. Data were analysed using t-tests and analysis of variance. A total of 313 women were followed up. American women who had immediate expander/implant surgery were compared with Swedish patients, whilst Americans who had undergone transverse rectus abdominis myocutaneous (TRAM) flap reconstructions were compared with Canadians. Women benefited from having breast reconstruction, but this improvement was not dependent upon country of origin. Swedish women reported less improvement in one subscale, that of general health, compared with American women (P=0.01). There were no cultural differences detected between Americans and Canadians. PMID- 17399660 TI - Post-mastectomy reconstruction: a comparative analysis on psychosocial and psychopathological outcomes. AB - Although multiple benefits have been reported from post-mastectomy reconstruction and particularly from an immediate procedure, limited psychiatric evaluation has been published. The present study was planned to evaluate the psychosocial and psychopathological outcome in patients who had undergone post-mastectomy breast reconstruction. We also investigated if any psychopathological condition could significantly affect the benefits of the reconstructive procedure. We compared 33 breast-reconstructed patients with 33 patients with mastectomy alone and 33 healthy women. All women underwent a psychiatric interview with four questionnaire-based scales (SASS, QL-index, HAM-A, HAM-D) to assess social adaptation, quality of life, anxiety and depression. A 4-point scale evaluated reconstructed patients' satisfaction. After a year, there was no statistical difference in social, sexual relationships and quality of life among reconstructed patients and healthy women. Regarding anxiety, no statistical difference was found between reconstructed and mastectomy groups. Statistically different lower depression levels in the healthy group and higher levels in the mastectomy group were found. Depression in the reconstructed group decreased compared to the mastectomy group. Differences between timing and reconstructive techniques were not statistically validated. Eight patients were dissatisfied (24.2%). Immediate reconstructed patients who had pre-existing major depressive disorder were dissatisfied. Breast-reconstructed patients' quality of life, social and sexual relationships are not significantly different to those of healthy women. Anxiety does not decrease. Timing and techniques do not seem to influence, significantly, outcome and satisfaction. Pre-existing diagnosis of major depressive disorder may be a contraindication to immediate breast reconstruction. PMID- 17399661 TI - Abdominal strength after breast reconstruction using a free abdominal flap. AB - BACKGROUND: Today, breast reconstruction with autologous tissue is most commonly done either as a free muscle sparring TRAM flap or as a DIEP flap. Studies of donor site morbidity have shown an advantage in using the DIEP flap. However, this procedure might also be associated with an increased risk of flow related complications and it is also thought to be more demanding and time consuming. A few studies have evaluated the abdominal wall strength after dissection of a TRAM flap or a DIEP flap. However, these studies do not distinguish between the various types of free TRAM flaps and they also compare TRAM procedures preformed in an early period to DIEP procedures done in a later period. METHODS: We used an isokinetic dynamometer to measure concentric, eccentric and isometric abdominal muscle strength in 32 patients who had had a unilateral breast reconstruction with a free MS-2 (15) or a DIEP (17) flap in the year 2003. RESULTS: No significant reduction in muscle strength was observed for concentric or isometric muscle strength. However, significant lower eccentric muscle strength was found in the TRAM compared to the DIEP group (p=0.05). There was no significant difference in abdominal strength between the two flap groups at low to moderate work intensity (isometric/concentric). At the greatest work intensity (eccentric muscle strength) the patients reconstructed with a DIEP flap had a clinical small, but significant advantage over the patients reconstructed with a MS-2 TRAM flap. PMID- 17399662 TI - Management of infraumbilical vertical scars in DIEP-flaps by crossover anastomosis. AB - The deep inferior epigastric perforator (DIEP)-flap continues to be the standard treatment in microsurgical breast reconstruction. Reasons for the popularity of the DIEP-flap include the availability of a large amount of tissue for the reconstruction of large breasts, a reliable vascular anatomy and an aesthetically pleasing donor site scar. However, the DIEP-flap is not considered the optimal choice as the donor tissue in all patients. Previous abdominal surgeries with resulting scars may threaten the success of a free DIEP-flap due to compromised vascularity within the flap. We elaborated a technique to increase the safety of breast reconstruction with the DIEP-flap in the presence of an infraumbilical vertical scar. After raising the DIEP-flap in a traditional manner on one side with harvesting of a considerate length of the inferior epigastric vessels, a segment of the superior epigastric vessels is left attached to the main pedicle. This stump of the superior epigastric vessels is now anastomosed under the microscope to a paraumbilical perforator on the contralateral side of the flap for in-flap microvascular augmentation. The above-mentioned technique was applied in five patients who presented with an infraumbilical vertical scar and were reconstructed with a DIEP-flap because of breast cancer. In three of the five patients there was an additional risk factor present such as smoking or diabetes mellitus. In all five patients no major complication due to marginal perfusion of the contralateral side of the flap was encountered. In two patients there was minor breakdown of fatty tissue that was managed conservatively in both cases. In flap microvascular augmentation of DIEP-flaps is a valuable tool for the plastic surgeon in microvascular breast reconstruction. It permits usage of the lower abdominal tissue even if perfusion is compromised due to midline scarring. We recommend this technique as a safe alternative in patients seeking autologous breast reconstruction in the presence of a midline abdominal scar. PMID- 17399663 TI - Delayed breast reconstruction using a combination of latissimus dorsi muscle flap and tissue expander with embodiment injection site. AB - This article presents our experience in delayed breast reconstruction, using a combination of latissimus dorsi muscle flap (LDMF) and tissue expander with embodiment injection site, in a two-stage approach. A consecutive sample of 50 patients was studied. The average length of follow up was 4 years (range 12-72 months). In all patients the above two-stage approach was performed. The results are encouraging. Using LDMF and tissue expansion for delayed breast reconstruction has proven to be useful. This approach provides sufficient muscular coverage of the implant. The level of patient satisfaction is high. Major complications are rare. The minor complications are represented mainly by the dorsal seroma. The technique with LDMF has been improved substantially during the past few years. In our experience, it provides the plastic surgeon with an excellent, safe and consistently successful method for delayed breast reconstruction. PMID- 17399664 TI - Formation of a reliable capsular flap in a rat model. AB - Prefabrication can be used to produce capsular flaps; other researchers have confirmed the feasibility of such flaps. Before the possibilities of capsular flaps can be explored, a reliable method to create these flaps has to be established first. METHODS: To produce capsular flaps in a rat model, the femoral vascular bundle was sandwiched between two silicone sheets. Three different methods were used and described. The capsule that formed between the two silicone sheets receives its main blood supply from that vascular pedicle. In this way pedicled capsular flaps were created. These flaps were used as a carrier for a skin graft, thus pre-laminating them, to test their ability for reconstructive surgery. The results of the three different methods of creating capsular flaps in a rat model were described and their results were evaluated. Especially the amount of capsule formation and the viability of the skin grafts was observed and compared. The feasibility of pre-laminated capsular flaps is confirmed and the most reliable method of creating them is described. PMID- 17399665 TI - Macroporous gelatine spheres as culture substrate, transplantation vehicle, and biodegradable scaffold for guided regeneration of soft tissues. In vivo study in nude mice. AB - In the course of development of a new type of filler for the correction of small defects in soft tissues we studied macroporous gelatine spheres as culture substrate, transplantation vehicle, and biodegradable scaffold for guided regeneration of soft tissues in vivo. We injected intradermally in nude mice gelatine spheres that had either been preseeded with human fibroblasts or preadipocytes, or left unseeded. We compared the extent of regenerated tissue with that found after injections of saline or single-cell suspensions of human fibroblasts or preadipocytes. Routine histological examinations and immunohistochemical staining for von Willebrand factor (indicating neoangiogenesis) were made after 7, 21, and 56 days. Injected saline or single cell suspensions had no effect. However, a quick and thorough tissue regeneration with developing neoangiogenesis was elicited by the gelatine spheres and the effect of spheres preseeded with preadipocytes surpassed the effect of spheres preseeded with fibroblasts, which in turn surpassed the effect of unseeded gelatine spheres. We suggest that minor soft tissue defects such as wrinkles or creases can be corrected by injection of naked macroporous gelatine spheres, whereas larger defects are best corrected by injection of macroporous gelatine spheres preseeded with fibroblasts, or preadipocytes, or both. PMID- 17399666 TI - A reverse-flow composite flap in the rat. AB - Reverse-flow flaps are currently particularly used for the reconstruction of defects of the distal part of the extremities. Despite their common usage there have been many reports of postoperative complications, especially resulting in partial or total flap necrosis. There is insufficient knowledge of flap haemodynamics, physiology and wound healing properties in reverse-flow flaps. Development of the proper experimental models is needed to investigate these issues. The purpose of this study was to describe a new reverse-flow flap model in the rat. A total of 20 adult Wistar rats weighing 200-250 g were used in this experiment. In five rats, the vascular anatomy of the auricle of the rat was determined by anatomic dissection and microangiography. In the experimental group (N=5), 1x1 cm reverse-flow composite flaps were harvested as a semi-island shape, based on the distal course of the medial branch of the anterior auricular artery. In the control group, consisting of five rats, the flap was designed and raised based on the proximal course of the medial auricular artery, again in a semi island shape. In the remaining five animals, a square-shaped composite tissue of the whole layer of the auricle, 1x1 cm in size, was harvested dividing all the bases circumferentially. The composite tissue was replaced in situ. While the former was considered a conventional antegrade-flow flap subgroup, the latter was designated as a graft subgroup. All flaps were replaced in situ. The survival of the flap was evaluated on postoperative day 7 by direct observation and microangiography. The skin island of all the reverse-flow flaps and conventional antegrade-flow flaps survived completely giving a success rate of 100%, whereas all grafts in the control group underwent complete necrosis. Microangiographic studies revealed the vascularity of the reverse-flow and antegrade-flow flaps, identifying the course of the auricular arteries. In conclusion, with its evident advantages of easy to design and harvesting, reliable survival pattern and consistent vascular structure, our new flap model will provide a means for future studies on flap haemodynamics, physiology in reverse-flow flaps. PMID- 17399667 TI - Beneficial effects of caffeic acid phenethyl ester (CAPE) on the ischaemia reperfusion injury in rat skin flaps. AB - BACKGROUND: Reperfusion injury is a phenomenon that occurs when tissues are subjected to ischaemia for a variable period of time, after which they are reperfused. Many factors have been implicated in the cause of reperfusion injury including free radicals and neutrophils. Caffeic acid (3,4-dihydroxycinnamic acid) phenethyl ester (CAPE) is an active component of propolis from honeybee; it has anti-inflammatory and immunomodulatory properties, and protective effects against ischaemia-reperfusion (I/R) injury. We investigated the effects of CAPE on the survival of skin flaps in the rat. MATERIALS AND METHODS: Eighteen Wistar rats were used, and randomly divided into three groups (n=6 rats each group): the control group (Group 1), ethanol group (Group 2), and CAPE group (Group 3). A caudally based rectangular flap, 3x10 cm in size, was elevated on the back of the rat, according to the method described by Khouri and colleagues. The flap was sutured back into its original place. In the control group, saline 0.2 ml/day was given intraperitoneally (i.p.). Five percent ethanol 0.2 ml/day was administered i.p. in the ethanol group, and CAPE 50 micromol/kg/day i.p. in the CAPE group. To observe the effects of CAPE, levels of malondialdehyde (MDA), nitric oxide (NO), glutathione (GSH), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) were measured from extracted skin tissue. Flap viability was evaluated seven days after the initial operation, measuring necrotic areas of flaps and total flap areas. RESULTS: MDA and NO levels were significantly decreased in CAPE group; and however, GSH, GSH-Px, and SOD enzyme activities were significantly increased in CAPE group. We believed that the CAPE had beneficial effects to improve the survival of skin flaps since it has antioxidative and anti inflammatory properties, and protective effects against I/R injury. PMID- 17399668 TI - Commentary on: A simple classification for standardisation of nomenclature in free flap outcome, by R. Srikanth, D. Mukunda Reddy, A. Mosahebi, J Plast Reconstr Aesthetic Surg 2006;59:1318-24. PMID- 17399669 TI - Salvage of an ischaemic 'kite flap' by an arterial supercharge: a case report. PMID- 17399670 TI - Letter in reply to "Akthar N, Smith MJ, McKirdy S, Page RE. Surgical delay in the management of dog bite injuries in children, does it increase the risk of infection?" J Plast Reconstr Aesthetic Surg 2006; 59(1):80-5. PMID- 17399671 TI - Cross infection risk in surgical marking pens. PMID- 17399672 TI - Drainage of large or symptomatic seromas: let the vacuum do the work. PMID- 17399673 TI - A helpful trick for the abdominoplasty scar. PMID- 17399674 TI - Long-term effect of optic edge design in a silicone intraocular lens on posterior capsule opacification. AB - PURPOSE: To compare the posterior capsule opacification (PCO) inhibiting effect of the round anterior and sharp posterior optic edge profile of the Clariflex silicone intraocular lens (IOL) [AMO Inc, Santa Ana, California, USA] with that of the double-round edge profile of the SI40 (Phacoflex; AMO Inc, Santa Ana, California, USA) silicone IOL over a period of three years. DESIGN: Prospective, randomized, double-masked, bilateral clinical trial. METHODS: The study took place at the Department of Ophthalmology, Medical University of Vienna, Austria. Fifty-two patients with age-related cataracts (104 eyes) were included in the study. Each patient received an SI40 IOL (round edges) in one eye and a Clariflex IOL with OptiEdge (sharp posterior optic edge) in the other eye. Follow-up examinations were at one week, one month, six months, and one, two, and three years. Digital retroillumination images were taken of each eye. The amount of PCO was subjectively assessed with the slit-lamp and objectively assessed by automated image analysis software (the computer program Automated Quantification of After-Cataract [AQUA]; Vienna, Austria) one, two, and three years after surgery. RESULTS: The Clariflex lens showed markedly less PCO at one, two, and three years after surgery. The mean AQUA PCO score was 1.39 for the SI40 and 0.56 for the Clariflex lens after one year, estimated at 1.64 and 0.57 after two years, and at 2.04 and 0.64, respectively, after three years (scale zero to 10; P < .001). The Nd:YAG laser capsulotomy rate was far higher in the SI40 group (Five cases vs one case at three years). CONCLUSIONS: The sharp posterior optic edge profile of the Clariflex silicone IOL led to marked and consistently less PCO than the round-edged SI40 IOL one, two, and three years after surgery. PMID- 17399675 TI - Three-year incidence and cumulative prevalence of retinopathy: the atherosclerosis risk in communities study. AB - PURPOSE: To describe the three-year incidence and cumulative prevalence of retinopathy and its risk factors. DESIGN: Population-based, prospective cohort study in four US communities. METHODS: In the Atherosclerosis Risk in Communities (ARIC) Study, 981 participants had retinal photography of one randomly selected eye at the third examination (1993 to 1995) and three years later at the fourth examination (1996). Photographs were graded on both occasions for retinopathy signs (for example, microaneurysm, retinal hemorrhage, and/or cotton-wool spots). Incidence was defined as participants without retinopathy at the third examination who developed retinopathy at the fourth examination, and cumulative prevalence was defined to include incident retinopathy as well as participants who had retinopathy at both the third and fourth examinations. RESULTS: The three year incidence and cumulative prevalence of any retinopathy in the whole cohort was 3.8% and 7.7%, respectively. In multivariable analysis, incident retinopathy was related to higher mean arterial blood pressure (odds ratio [OR] 1.5, 95% confidence interval [CI] 1.0 to 2.3, per standard deviation increase in risk factor levels), fasting serum glucose (OR 1.6, 95% CI 1.3 to 2.1), serum total cholesterol (OR 1.4, 95% CI 1.0, 2.0), and plasma fibrinogen (OR 1.4, 95% CI 1.1 to 1.9). Among persons without diabetes, the three-year incidence and cumulative prevalence of nondiabetic retinopathy was 2.9% and 4.3%, respectively. Incident nondiabetic retinopathy was related to higher mean arterial blood pressure (OR 1.4, 95% CI 0.9 to 2.3) and fasting serum glucose (OR 1.5, 95% CI 1.0 to 2.3). Among persons with diabetes, the three-year incidence and cumulative prevalence of diabetic retinopathy was 10.1% and 27.2%, respectively. CONCLUSIONS: Retinopathy signs occur frequently in middle-aged people, even in those without diabetes. Hypertension and hyperglycemia are risk factors for incident retinopathy. PMID- 17399677 TI - Molecular haplotyping of tandem single nucleotide polymorphisms by allele specific PCR. AB - BARD1 Val507Met (1592A>G) is an interesting marker for association studies on cancer risk. However, studies are scarce in the literature, probably reflecting the methodological problem imposed by the fact that next to the 1592A>G stands the 1591C>T single nucleotide polymorphism (SNP). We have designed an allele specific PCR method capable of molecular haplotyping tandem SNPs. In the tandem SNPs haplotyping assay (tSNPh), four reverse primers are designed to be perfect matches of each potential haplotype. The forward primer is labeled with a fluorochrome. PCR products are analyzed by capillary electrophoresis. Haplotyping is performed by size calling. To ascertain the accuracy and reproducibility of the assay, we measured the level of concordance with sequencing data in 124 samples. In vitro-generated templates have been used for further testing. We developed a novel and reliable assay that permits typing two SNPs directly adjacent to each other, avoiding mutual interferences. The method is amenable to automation and high throughput. We expect that this assay will contribute to clarifying the role of BARD1 in cancer susceptibility. In addition, we suggest that tandem SNPs are potentially interesting polymorphic markers in which molecular haplotyping can be performed easily. PMID- 17399676 TI - Severity of visual field loss and health-related quality of life. AB - PURPOSE: To examine the association between severity of visual field loss (VFL) and self-reported health-related quality of life (HRQOL) in a population-based sample. DESIGN: Population-based cross-sectional study. METHODS: Participants in the Los Angeles Latino Eye Study (LALES) underwent a comprehensive ophthalmic examination including visual field testing by the Humphrey Automated Field Analyzer II (Swedish Interactive Thresholding Algorithm [SITA] Standard 24-2) [Carl Zeiss Meditec, Dublin, California, USA]. Mean deviation (MD) scores were used to determine severity of VFL both as a continuous variable and stratified by severity: no VFL (MD >or= -2 decibels [dB]), mild VFL (-6 dB < MD < -2 dB), and moderate to severe VFL (MD < -6 dB). HRQOL was assessed by the Medical Outcomes Study 12-item Short-Form Health Survey (SF-12) and the National Eye Institute Visual Function Questionnaire (NEI-VFQ-25). Linear regression analyses and analysis of covariance were used to assess the relationship between HRQOL scores and VFL. RESULTS: Of the 5,213 participants included in this study, 18% had unilateral mild, 1.5% unilateral moderate to severe, 19% bilateral mild, and 6.5% bilateral moderate to severe VFL. Worse NEI-VFQ-25 and SF-12 HRQOL scores were associated with VFL in a linear manner. Four- to 5-dB differences in VFL were associated with a five-point difference in the NEI-VFQ-25 composite and most subscale scores. Persons with VFL had the greatest difficulty with driving activities, dependency, mental health, distance vision, and peripheral vision. CONCLUSIONS: HRQOL is diminished even in persons with relatively mild VFL on the basis of MD scores. Prevention and management of persons with VFL may be important in preventing or reducing poor HRQOL related to difficulties in driving, distance and peripheral vision activities, and a sense of dependency. PMID- 17399679 TI - The human cardiac hormone fragment N-terminal pro B-type natriuretic peptide is an intrinsically unstructured protein. AB - The cardiac hormone B-type natriuretic peptide (BNP) is synthesized as a prepro 134 residue molecule which is further proteolytically processed into a 76 residue fragment termed N-terminal proBNP (NT-proBNP) and the active portion of this hormone, a 32-residue disulfide-linked peptide (BNP-32). The active hormone regulates cardiac hemodynamic output while as yet no biological function has been attributed to NT-proBNP. Some solution properties of synthetically generated NT proBNP in benign media are known. The protein is monomeric, elutes aberrantly on size-exclusion chromatography as an apparent larger molecular species, and possesses little global secondary structure as assessed by circular dichroism. To explore the solution structure of NT-proBNP in greater detail, we use 2D-NOESY and 2D-TOCSY NMR on recombinant NT-proBNP to obtain a high resolution solution conformation at the alpha-carbon level. Importantly, NH(i)-NH(i+1) coupling is virtually absent at room temperature implying that large stretches of primary sequence are unordered. Together, the results of these physicochemical measurements classify NT-proBNP as a naturally unfolded protein referred to as an Intrinsically Unstructured Protein (IUP). The calculations of FoldIndex, a computer program which predicts disorder, were compared to the experimental results described here for NT-proBNP in addition to proBNP. NT-proBNP thus appears to be an ideal candidate for the study of native, unfolded proteins. PMID- 17399680 TI - Failures to retrieve specific memories in previously depressed individuals: random errors or content-related? AB - Failures to retrieve specific memories have been related to general tendencies to commit errors in executive functioning tasks, raising the question of whether they are simply a reflection of randomly occurring lapses in cognitive control or whether their occurrence is in any way related to memory content. This study investigated the relation between failures to retrieve specific autobiographical memories and dysfunctional attitudes in individuals at risk for depression and controls. Sixteen previously depressed and 19 never-depressed participants were assessed for dysfunctional attitudes regarding "need for approval" and "performance evaluation". One week later, they were asked to generate specific memories following dependency- and achievement-related cue words, both under single and dual task conditions. In previously depressed participants, dysfunctional attitudes regarding "need for approval" significantly predicted specificity for dependency-related events under dual task conditions. The findings provide qualified support for a role of content and demonstrate how, in previously depressed individuals, content effects can surface when cognitive control is undermined. PMID- 17399681 TI - Characterization of wheat germin (oxalate oxidase) expressed by Pichia pastoris. AB - High-level secretory expression of wheat (Triticum aestivum) germin/oxalate oxidase was achieved in Pichia pastoris fermentation cultures as an alpha-mating factor signal peptide fusion, based on the native wheat cDNA coding sequence. The oxalate oxidase activity of the recombinant enzyme is substantially increased (7 fold) by treatment with sodium periodate, followed by ascorbate reduction. Using these methods, approximately 1 g (4x10(4) U) of purified, activated enzyme was obtained following eight days of induction of a high density Pichia fermentation culture, demonstrating suitability for large-scale production of oxalate oxidase for biotechnological applications. Characterization of the recombinant protein shows that it is glycosylated, with N-linked glycan attached at Asn47. For potential biomedical applications, a nonglycosylated (S49A) variant was also prepared which retains essentially full enzyme activity, but exhibits altered protein-protein interactions. PMID- 17399682 TI - Marinobufagenin interferes with the function of the mineralocorticoid receptor. AB - Marinobufagenin (MBG) is a cardiotonic steroid of the bufadienolide class of compounds which has the ability to inhibit the ubiquitous enzyme, Na+/K+-ATPase, resulting in natriuresis. The involvement of MBG in the pathogenesis of volume expansion-mediated forms of hypertension has been suggested for some time, and we have proposed that MBG participates in the hypertension noted in preeclampsia. We examined the hypothesis that MBG might contribute to these forms of hypertension by promoting the activity of the mineralocorticoid receptor (MR). However, our data demonstrate that instead, MBG interferes with the functioning of the MR by inhibiting the transcriptional activity of the receptor, and this is reflected in a reduced interaction between the SRC-3 coactivator and the MR. Thus, the ability of MBG to cause a natriuresis may be due, not only to inhibition of Na+/K+-ATPase activity, but also to its ability to interfere with MR-dependent expression of the Na/K/H exchanger in the late distal nephron. PMID- 17399683 TI - Promotion of attachment of human bone marrow stromal cells by CCN2. AB - Cell attachment is a crucial step in tissue regeneration. In this study, human bone marrow stromal cells (hBMSCs) were isolated, and the effects of CCN2 on their attachment were examined. CCN2 significantly enhanced the hBMSC attachment, and this enhanced cell attachment was mainly regulated by the C-terminal module of CCN2. This enhancement was negated by the anti-integrin alpha(v)beta(3) antibody and p38 MAPK inhibitor, and phosphorylation of p38 MAPK was detected upon the enhanced cell attachment mediated by CCN2. We thus conclude that CCN2 enhances hBMSC attachment via integrin-p38 MAPK signal pathway. Enhanced hBMSC attachment on hydroxyapatite plates by CCN2 further indicated the utility of CCN2 in bone regeneration. PMID- 17399684 TI - Sequence and expression analysis of histone deacetylases in rice. AB - Histone acetylation levels are determined by the action of histone acetyltransferases and histone deacetylases (HDACs). Sequence similarity and profile searching tools were used to analyze the genome sequence of rice (Oryzae sativa) for genes encoding HDAC proteins. The rice RPD3/HDA1-family HDAC proteins can be divided into four classes based on sequence similarity and phylogenetic analysis of sequences obtained from the rice genome. The spatial expression pattern of rice HDACs genes indicated that some HDAC genes have different expression profiles. Furthermore, our analysis indicated that expression of HDA705, HDT701, and HDT702 could be affected by salicylic acid, jasmonic acid or abscisic acid. Expression of HDA714, SRT702, and SRT701 could be modulated by abiotic stresses, such as cold, mannitol and salt. These results indicate that different HDAC genes have distinct expression patterns and members of rice HDAC families may be involved in plant response to environmental stresses. PMID- 17399685 TI - Induction of human adiponectin gene transcription by telmisartan, angiotensin receptor blocker, independently on PPAR-gamma activation. AB - Adiponectin, an adipose tissue-specific plasma protein, has been shown to ameliorate insulin resistance and inhibit the process of atherosclerosis. Recently, several reports have stated that angiotensin type 1 receptor blockers (ARBs), increase adiponectin plasma level, and ameliorate insulin resistance. Telmisartan, a subclass of ARBs, has been shown to be a partial agonist of the peroxisome proliferator-activated receptor (PPAR)-gamma, and to increase the plasma adiponectin level. However, the transcriptional regulation of the human adiponectin gene by telmisartan has not been determined yet. To elucidate the effect of telmisartan on adiponectin, the stimulatory regulation of human adiponectin gene by telmisartan was investigated in 3T3-L1 adipocytes, utilizing adenovirus-mediated luciferase reporter gene-transferring technique. This study indicates that telmisartan may stimulate adiponectin transcription independent of PPAR-gamma. PMID- 17399686 TI - TAp63gamma regulates hOGG1 and repair of oxidative damage in cancer cell lines. AB - We showed that TAp63gamma regulates hOGG1. Using chromatin immunoprecipitation (ChIP), we found that TAp63gamma binds to the hOGG1 promoter. Reintroduction of wild-type TAp63gamma into HEK 293 cells, induced transcription of hOGG1 promoter, leading to increase in RNA and protein. Using RNAi studies, we observed that TAp63gamma-RNAi resulted in reduced hOGG1 RNA and protein in HeLa cells. This decrease in hOGG1 expression was associated with reduced cell viability upon oxidative damage. Taken together, our results indicate that hOGG1 is a direct target of TAp63gamma, suggesting a role for TAp63gamma in oxidative damage and repair. PMID- 17399687 TI - Activation of a novel pathway involving Mms1 and Rad59 in sgs1 cells. AB - Unequal sister chromatid recombination (uSCR) is elevated in budding yeast sgs1 mutants, which lack a homolog of the human BLM gene that causes Bloom syndrome. Examination of the mechanism responsible for elevated uSCR in sgs1 mutants showed that mutation of RAD51 also resulted in hyper-uSCR. Data from this study show that defects in the Rad51-Sgs1-dependent and Sgs1-dependent lesion-bypass pathways activate Rad59-Rad1- and Rad59-dependent pathways, respectively, resulting in uSCR. Moreover, the elevation of uSCR in sgs1 and rad51 mutants was dependent on MMS1, which encodes one of the components of the Mms22 module. Lastly, a putative role of Mms1 in the elevation of uSCR and a possible mechanism by which uSCR is elevated as a result of defective Sgs1 and Rad51 are discussed. PMID- 17399688 TI - Evidence of P-body-like structures in Trypanosoma cruzi. AB - Gene expression in trypanosomatids is mainly regulated post-transcriptionally. One of the mechanisms involves the differential stability of mRNAs. However, the existence of other mechanisms involving the accessibility of mRNAs to the translation machinery cannot be ruled out. Defined cytoplasmic foci containing non-translating mRNPs, known as P-bodies, have been discovered in recent years. P bodies are sites where mRNA can be decapped and 5'-3' degraded or stored for subsequent return to polysomes. The highly conserved DEAD box helicase Dhh1p is a marker protein of P-body functions. Here, we report the identification and cloning of a Trypanosoma cruzi Dhh1 homolog gene. TcDhh1 expression is not regulated through the parasite life cycle or under stress conditions. We show that TcDhh1 is present in polysome-independent complexes and is localized to discrete cytoplasmic foci, resembling P-bodies; these foci vary in number according to nutritional stress conditions and cycloheximide/puromycin treatment. PMID- 17399689 TI - Role of asymmetric dimethylarginine in homocysteine-induced apoptosis of vascular smooth muscle cells. AB - Homocysteine (Hcy) could induce apoptosis of vascular smooth muscle cells (VSMC). Asymmetric dimethylarginine (ADMA) has been thought as a novel risk factor for cardiovascular diseases. We hypothesized that ADMA mediates homocysteine-induced apoptosis of VSMC. In this experiment the level of ADMA in the medium measured by high-performance liquid chromatography (HPLC) was elevated when the apoptosis of T/G HA-VSMC was induced by Hcy which was detected by Hoechst33342 staining or flow cytometry (FCM) with Annecin V+Propidium Iodide (PI). Exogenous ADMA induced the apoptosis of VSMC. At the same time, ADMA elevated the level of intracellular reactive oxidative species (ROS) determined by fluorescent ROS detection kit. The activation of JNK and p38MAPK contributed to ADMA-induced apoptosis of VSMC. The present results suggest that endogenous ADMA is involved in apoptosis of VSMC induced by Hcy, and the effects of ADMA is related to elevation of intracellular ROS and activation of JNK/p38MAPK signaling pathways. PMID- 17399690 TI - Visual recognition memory and auditory brainstem response in infant rhesus monkeys exposed perinatally to environmental tobacco smoke. AB - The impact of perinatal exposure to environmental tobacco smoke (ETS) on cognitive development is controversial. We exposed rhesus monkeys to ETS or filtered air (5 animals per group) beginning in utero on day 50 of pregnancy and continuing throughout postnatal testing. In infancy, we evaluated both groups for visual recognition memory and auditory function (auditory brainstem response). The ETS group showed significantly less novelty preference in the visual recognition task whereas no effects on auditory function were detected. These preliminary results support the view that perinatal ETS exposure has adverse effects on cognitive function and indicate further that rhesus monkeys may provide a valuable nonhuman primate model for investigating this link. PMID- 17399692 TI - Serum-withdrawal-dependent apoptosis of hippocampal neuroblasts involves Ca++ release by endoplasmic reticulum and caspase-12 activation. AB - Apoptotic death caused by diseases or toxic insults is preceded and determined by endoplasmic reticulum dysfunction and altered intraluminar calcium homeostasis in many different cell types. With the present study we have explored the possibility that the ER stress could be involved also in apoptotic death induced by serum deprivation in neuronal cells. We have chosen as a model of study the cell line HN9.10e, constituted by immortalized hippocampal neuroblasts. The Ca(++) concentration in the lumen of the ER has been evaluated by using the low affinity Ca(++) probe Mag-fluo-4. We show that serum deprivation lowers the ER Ca(++) concentration with a time course closely related to the increase of apoptosis incidence. Serum deprivation also enhances the expression of a well known marker of ER stress, the glucose-regulated protein-78 (GRP-78), a member of the heat shock/stress response protein family. Moreover, in serum-deprived neuroblasts, following GRP-78 up-regulation, the ER-associated procaspase-12 is cleaved with a time course which parallels the ER calcium loss while activation of caspase-3 is a later event. Depletion of ER Ca(++) by thapsigargin, a specific inhibitor of the ER-associated Ca(++) ATPase, also produces caspase-12 processing and apoptotic cell death, whereas agents capable of reducing the ER calcium loss protect the cells from serum-deprivation-induced apoptosis. These findings indicate that, in hippocampal neuroblasts, Ca(++) mobilization from ER and caspase-12 activation are components of the molecular pathway that leads to apoptosis triggered by serum deprivation and may constitute an amplifying loop of the mitochondrial pathway. PMID- 17399691 TI - Prenatal choline deficiency increases choline transporter expression in the septum and hippocampus during postnatal development and in adulthood in rats. AB - Supplementation of maternal diet with the essential nutrient, choline, during the second half of pregnancy in rats causes long-lasting improvements in spatial memory in the offspring and protects them from the memory decline characteristic of old age. In contrast, prenatal choline deficiency is associated with poor performance in certain cognitive tasks. The mechanism by which choline influences learning and memory remains unclear; however, it may involve changes to the hippocampal cholinergic system. Previously, we showed that the hippocampi of prenatally [embryonic days (E) 11-17] choline-deficient animals have increased synthesis of acetylcholine (ACh) from choline transported by the high-affinity choline transporter (CHT) and reduced ACh content relative to the control and to the E11-17 choline-supplemented rats. In the current study, we found that, during postnatal period [postnatal days (P) 18-480], prenatal choline deficiency increased the expression of CHT mRNA in the septum and CHT mRNA and protein levels in the hippocampus and altered the pattern of CHT immunoreactivity in the dentate gyrus. CHT immunoreactivity was more prominent in the inner molecular layer in prenatally choline-deficient rats compared to controls and prenatally choline-supplemented animals. In addition, in all groups, we observed a population of hilar interneurons that were CHT-immunoreactive. These neurons are the likely source of the hippocampal CHT mRNA as their number correlated with the levels of this mRNA. The abundance of hippocampal CHT mRNA rose between P1 and P24 and then declined reaching 60% of the P1 value by P90. These data show that prenatal availability of choline alters its own metabolism (i.e., CHT expression). While the upregulated CHT expression during the period of prenatal choline deficiency may be considered as a compensatory mechanism that could enhance ACh synthesis when choline supply is low, the persistent upregulation of CHT expression subsequent to the brief period of prenatal deprivation of choline in utero might be beneficial during choline deficiency in adulthood. PMID- 17399693 TI - Post-infarction remodeling is independent of mitogen-activated protein kinase kinase 3 (MKK3). AB - OBJECTIVES: Our aim was to examine the role of mitogen-activated protein kinase kinase 3 (MKK3) in the development of left ventricular (LV) remodeling following myocardial infarction (MI). METHODS: MKK3-null mice were subjected to permanent coronary artery ligation. Twenty-eight days after MI, haemodynamics in male mkk3+/+(WT) and mkk3-/-(KO) littermates were assessed using a pressure conductance catheter. MI groups were compared to un-operated time-matched WT and KO controls. RESULTS: MI caused significant LV contractile dysfunction and dilatation which did not differ by genotype. Detailed morphometric analysis of excised hearts confirmed these similar global indices of remodeling and also demonstrated that pathological changes within remote myocardium and scar did not differ between KO and WT hearts. CONCLUSIONS: Despite numerous lines of evidence suggesting MKK3 is the relevant kinase upstream of p38 mitogen-activated protein kinase in LV remodeling these processes can continue in its absence. PMID- 17399694 TI - Efficacy of DL-alpha-lipoic acid on methanol induced free radical changes, protein oxidative damages and hsp70 expression in folate deficient rat nervous tissue. AB - DL-alpha-Lipoic acid (LPA) was reported to be effective in reducing free radicals generated by oxidative stress. The protective of effect of LPA on methanol (MeOH) induced free radical changes and oxidative damages in discrete regions of rat brain have been reported in this study. Folate deficient rat (FDD) model was used. The five animal groups (saline control, FDD control, FDD+MeOH, FDD+LPA+MeOH, LPA control) were used. The FDD+MeOH and FDD+LPA+MeOH animals were injected intraperitoneally with methanol (3gm/kg). After 24h, the level of free radical scavengers such as, superoxide dismutase, catalase, glutathione peroxidase, reduced glutathione was estimated in six discrete regions of brain, retina and optic nerve. Level of protein thiol, protein carbonyl and lipid peroxidation was also estimated. Expression of heat shock protein 70 mRNA (hsp70) was studied in the cerebellum and hippocampus by reverse transcriptase PCR. All the samples showed elevation in the level of free radical scavenging enzymes and reduced level of glutathione in the FDD+MeOH group in relation to the other groups. hsp70 expression was more in FDD+MeOH group when compared to FDD+LPA+MeOH group. In conclusion, MeOH exposure leads to increased free radical generation and protein oxidative damages in the rat nervous tissue. Treatment with LPA prevents oxidative damage induced by MeOH exposure. PMID- 17399695 TI - Cytotoxicity of zinc-containing bioactive glasses in contact with human osteoblasts. AB - Bioactive glasses such as Hench's 45S5 have applications to tissue engineering and bone repair: the insertion of zinc has been proposed to improve their bone bonding ability and to slacken their dissolution in extracellular body fluids. In view of a potential clinical application, we have investigated whether zinc containing 45S5 (HZ) glasses might be cytotoxic for human MG-63 osteoblasts. In our experimental conditions, after 24h of incubation HZ glasses released significant amounts of Zn(2+) and induced in MG-63 cells release of lactate dehydrogenase (index of cytotoxicity) and the following indexes of oxidative stress: (i) accumulation of intracellular malonyldialdehyde, (ii) increased activity of pentose phosphate pathway, (iii) increased expression of heme oxygenase-1, (iv) increased activity of Cu,Zn-superoxide dismutase, (v) decreased level of intracellular thiols. These effects were inversely related to the zinc content of glass powders, were mimicked by ZnCl(2) solutions and were prevented by either metal chelators (EDTA, NTA) or the antioxidant ascorbate, suggesting that Zn(2+) released fastly from HZ glasses can cause MG-63 cell damage via an oxidative stress. This work highlights the importance of designing Zn-containing bioactive glasses without cytotoxic effects and gives supplementary information about the prooxidant role of zinc in living systems. PMID- 17399696 TI - Diagnostic accuracy of human telomerase reverse transcriptase mRNA in malignant pleural effusions: A preliminary report for in situ hybridization detection. AB - BACKGROUND: Detection of human telomerase reverse transcriptase (hTERT) mRNA by in situ hybridization (ISH) may be valuable in the diagnosis of cancer. We assessed the diagnostic performance of hTERT mRNA in cells from pleural fluid in malignant pleural effusions (PEs). METHODS: We used a 2-step ISH with digoxin labelled oligonucleotide probes to detect hTERT mRNA in a blinded prospective study of cells from 103 unselected pleural fluid specimens. The reference standard for malignant PEs was clinical evaluation and pleural fluid cytology, combined with pleural biopsy, other examination and follow-up as needed. RESULTS: According to the final diagnoses, there were 41 malignant PEs, 55 benign PEs and 7 cases with uncertain etiology. When the 7 cryptogenic cases were excluded, the sensitivity and specificity of detectable hTERT mRNA for malignancy were 80% and 95%, respectively. When detection of hTERT mRNA was combined with clinical repeated pleural fluid cytology, the sensitivity and specificity were 90% and 95%, respectively. CONCLUSIONS: Detection of hTERT mRNA in cells from pleural fluid by ISH could potentially be used in diagnosing malignant PEs as an aid. Further investigations with stricter controls and cross-validation tests will be warranted. PMID- 17399697 TI - Biomarkers of liver fibrosis: clinical translation of molecular pathogenesis or based on liver-dependent malfunction tests. AB - BACKGROUND: Fibrosis is the excessive deposition and histological redistribution of extracellular matrix (ECM) in the tissue as consequence of chronic liver damage. It leads to progressive liver insufficiency, portal hypertension and ultimately to cirrhosis and primary liver cell carcinoma. There is a strong demand for reliable, organ- and disease-specific, non-invasive biomarkers of fibrosis and fibrogenesis to replace or to complement the invasive method of needle biopsy, which is afflicted with a high degree of sampling error. METHODS: A systematic literature search was performed using electronic databases and reference lists of relevant publications to ascertain studies with non-invasive biomarkers of liver fibrosis. RESULTS: Two classes of serum biomarkers can be differentiated: Class I markers are those, which reflect ECM turnover (fibrogenesis and fibrolysis) and/or fibrogenic cell changes, mainly of hepatic stellate cells, which are the dominant profibrogenic cell type in liver. They are mostly cost intensive, single laboratory tests and derive from the translation of fibrogenic mechanisms into clinical application. Examples are procollagen peptides, hyaluronan, and laminin. Class II biomarkers are based on algorithmic evaluation of commonly observed functional alterations of the liver that do not necessarily reflect ECM metabolism and/or fibrogenic cell changes. About 20 numerical scores or indices are reported for parameters, which are mostly routine laboratory tests and frequently multiparametric (panels). Among them fibrotest, hepascore, ELF-score have reached limited clinical application. CONCLUSIONS: Up to now the impact of both classes of biomarkers for diagnosis and monitoring of fibrosis, fibrogenesis, and fibrolysis is limited. They cannot replace needle biopsy but some of them might be complementary in follow-up studies. Innovative methods like proteomics and glycomics to establish fibrosis-specific serum protein and glycosylation patterns, respectively, might have a high potential for diagnosis and monitoring of fibrogenesis. PMID- 17399698 TI - The on/off of Pax6 controls the tempo of neuronal differentiation in the developing spinal cord. AB - During neurogenesis, complex networks of genes act sequentially to control neuronal differentiation. In the neural tube, the expression of Pax6, a paired box-containing gene, just precedes the appearance of the first post-mitotic neurons. So far, its only reported function in the spinal cord is in specifying subsets of neurons. Here we address its possible function in controlling the balance between proliferation and commitment of neural progenitors. We report that increasing Pax6 level is sufficient to push neural progenitors toward cell cycle exit and neuronal commitment via Neurogenin 2 (Ngn2) upregulation. However, neuronal precursors maintaining Pax6(On) fail to perform neuronal differentiation. Conversely, turning off Pax6 function in these precursors is sufficient to provoke premature differentiation and the number of differentiated neurons depends of the amount of Pax6 protein. Moreover, we found that Pax6 expression involves negative feedback regulation by Ngn2 and this repression is critical for the proneural activity of Ngn2. We present a model in which the level of Pax6 activity first conditions the moment when a given progenitor will leave the cell cycle and second, the moment when a selected neuronal precursor will irreversibly differentiate. PMID- 17399699 TI - Effects of lipopolysaccharide on intestinal P-glycoprotein expression and activity. AB - It is well known that pharmacokinetics is often altered by changing the expression and activity of P-glycoprotein during sepsis. However, there have been few reports about expression and activity of P-glycoprotein in the small intestine during sepsis. We examined the levels of intestinal P-glycoprotein expression and activity using a rat sepsis model induced by lipopolysaccharide (LPS, from Escherichia coli). LPS was administered to male Wistar/ST rats intraperitonealy (i.p.) at 5 mg/kg. The small intestine was excised before and 1, 3 and 7 days after LPS administration, and the intestinal P-glycoprotein expression was determined using Western blot analysis. The activity of P glycoprotein was evaluated by measuring the efflux of rhodamine-123 (Rho123) in rats using an in situ single perfusion method. The changes of permeability via the paracellular route were evaluated by measuring the amount of fluorescein isothicyanate-dextran 4400 (FD-4) in a similar way. On Day 1 after LPS administration, both the level of P-glycoprotein expression and the total amount of Rho123 excreted into the intestinal lumen decreased significantly, but levels of both AUC2-95 and CLtot were not significantly different as compared with the control group. On Day 3, the total P-glycoprotein, including intestinal P glycoprotein, might have been induced by sepsis, and then the excretion of P glycoprotein substrate drugs into the intestinal lumen increased more than that of the control group. On Day 7, all pharmacokinetic parameters returned to the control level. Thus the intestinal P-glycoprotein function recovered within 3 days of LPS administration. PMID- 17399700 TI - Receptor tyrosine kinase inhibitors AG013764 and AG013711 reduce choroidal neovascularization in rat eye. AB - Age-related macular degeneration (AMD) is the major cause of blindness for people over 60. In the "wet" form of AMD compounds targeting growth factor signaling pathways such as VEGF have been a major focus for therapeutic interventions. In a previously developed rat model of CNV, we utilized two receptor tyrosine kinase inhibitors (RTKi) to block VEGFR-1, VEGFR-2 and PDGFR signaling following the establishment of CNV. AAV-VEGF(165) was injected into the subretinal space of rats at postnatal days 15-17. Six weeks later, a suspension of RTK inhibitors, AG013764 or AG013711, was injected intraperitoneally (IP, twice daily) or intravitreally (every five days) over a two week period. FITC-dextran whole mounts of RPE-choroid-sclera were prepared after the animals were sacrificed. CNV area was quantified using Neurolucida to measure the hyperfluorescence on FITC dextran whole-mounts. Histology and immunohistochemistry were performed as described previously. VEGF expression in control and treated eyes was confirmed by immunohistochemistry and histological sections indicated recovery of retinal morphology and CNV reduction in treated eyes. In the animals IP injected with AG013764 or AG013711 the mean CNV level was reduced by 25 to 33% compared to control, but this effect did not achieve statistical significance. Intravitreal injections of AG013764 or AG013711 reduced the level of CNV by approximately 60% compared to control (p<0.005 or p<0.05, respectively). These data show that two RTK inhibitors, AG013764 or AG013711, delivered intravitreally, significantly reduce blood vessel proliferation in this AAV-VEGF(165) model of CNV. PMID- 17399702 TI - Estrogen up-regulation of metalloproteinase-2 and -9 expression in rabbit lacrimal glands. AB - Increased levels of the matrix metalloproteinases (MMPs)-2 and -9 have been found in tear fluids of patients with dry eye disease, suggesting that these MMPs may be implicated in the pathogenesis of this disease. One of the main causes of dry eye disease is lacrimal gland insufficiency. However, the contribution of the lacrimal gland (LG) to the expression and production of MMP-2 and MMP-9 in tears is not known. Since dry eye disease occurs more frequently in women, sex hormones, especially estrogens, have also been implicated in the pathogenesis of this disease. Estrogens have been shown to regulate the synthesis levels of MMP-2 and MMP-9 in several tissues, Thus, the purpose of these studies was to determine if: (1) rabbit lacrimal glands secrete MMP-2 and MMP-9; (2) MMP-2 and MMP-9 are produced by lacrimal epithelial cells and/or lacrimal lymphocytes; and (3) the expression, activity and level of these enzymes are regulated by sex hormones. Lacrimal epithelial cells (LEC) and lacrimal lymphocytes (LL) from sexually mature New Zealand White female rabbits were isolated, purified and cultured with and without 10(-6)M dihydrotestosterone (DHT) or 10(-6), 10(-8), 10(-9) and 10( 10)M 17beta-estradiol (E2). The culture supernatants were analyzed by zymography and western blotting (WB) using polyclonal anti-human MMP-2 and MMP-9 antibodies. LGs were also collected from rabbits 7 days after being sham-operated, ovariectomized (OVX), OVX treated with 4 mg/kg DHT, and OVX treated with 0.5 mg/kg of E2. LGs were collected and processed for RNA extraction as well as protein determination using WB and immunocytochemistry. The pro-forms of MMP-2 and MMP-9 were detected in primary LEC and LL culture medium by zymography and WB. Pro-MMP-2 and pro-MMP-9 were also detected at the gene and protein levels in the lacrimal glands of all four treatment groups, with the highest levels and gene expression found in the estrogen-treated group. These results suggest that both pro-MMP-2 and pro-MMP-9 are secreted by the lacrimal gland and appear to be up-regulated by estrogen. The role of the lacrimal MMPs in the pathogenesis of dry eye disease needs to be further investigated. PMID- 17399703 TI - Information theory, single neurons and gamma oscillations in the human subthalamic nucleus. PMID- 17399701 TI - Assay of mucins in human tear fluid. AB - Mucin genes, both secreted (MUC2, MUC5AC, MUC5B, MUC7) and membrane associated (MUC1, MUC4, MUC16), have been reported to be expressed by ocular surface epithelia. The purpose of this study was to comprehensively assay the mucin content of human tear fluid using multiple antibodies for each mucin and to develop a sensitive, semi-quantitative method for the assay of mucins in tears. Tear washes were obtained by instillation of saline onto the ocular surface, followed by collection from the inferior fornix. Tear proteins were separated in 1% agarose gels, transferred to nitrocellulose membrane by vacuum blotting and probed with multiple antibodies recognizing MUC1, MUC2, MUC4, MUC5AC, MUC5B, MUC7 and MUC16. Binding was detected using chemiluminescence, and quantity was determined by densitometry. Serial dilutions of pooled tears from normal individuals were assayed to determine the linear range of detectability. MUC1, MUC4, MUC16, MUC5AC and low levels of MUC2 were consistently detected in human tear fluid, while MUC5B and MUC7 were not. Use of several antibodies recognizing different epitopes on the same mucin confirmed these findings. The antibodies to mucins bound to serial dilutions of tears in a linear fashion (r2 > 0.9), indicating the feasibility of semi-quantitation. MUC5AC in tear fluid had an increased electrophoretic mobility compared to MUC5AC isolated from conjunctival tissue. This study provides clear evidence that the mucin component of tears is a mixture of secreted and shed membrane-associated mucins, and for the first time demonstrates MUC16 in tear fluid. Immunoblots of tears using agarose gel electrophoresis and chemiluminescence detection provide a semi-quantitative assay for mucin protein that will be useful for comparisons with tears from diseased eyes or after pharmacological intervention. PMID- 17399704 TI - Clinical efficacy of antiparasite treatments against intestinal helminths and haematic protozoa in Gallotia caesaris (lizards). AB - Intestinal helminths and blood protozoa are common parasites of lizards. In captivity they can be a serious problem, but no information on treatments is available. In this study, several antiparasitic drugs were studied in Gallotia caesaris (Lacertida) from La Gomera, the Canary Islands, Spain. A lack of efficacy of chloroquine, and a decrease in haemogregarine (Apicomplexa) infection by atovaquone-proguanil was demonstrated. Furthermore, a positive effect of fenbendazole against intestinal nematode infection was observed. In the Canary Islands, Gallotia bravoana and Gallotia simonyi (Lacertida) are included in a captive breading recovery plan. The present results are useful in order to keep these animals in good condition and to control their parasites. Moreover, these results are important for reptiles kept as pets, due to the previous lack of knowledge of parasite management. PMID- 17399705 TI - The methylerythritol phosphate pathway for isoprenoid biosynthesis in coccidia: presence and sensitivity to fosmidomycin. AB - The apicoplast is a recently discovered, plastid-like organelle present in most apicomplexa. The methylerythritol phosphate (MEP) pathway involved in isoprenoid biosynthesis is one of the metabolic pathways associated with the apicoplast, and is a new promising therapeutic target in Plasmodium falciparum. Here, we check the presence of isoprenoid genes in four coccidian parasites according to genome database searches. Cryptosporidium parvum and C. hominis, which have no plastid genome, lack the MEP pathway. In contrast, gene expression studies suggest that this metabolic pathway is present in several development stages of Eimeria tenella and in tachyzoites of Toxoplasma gondii. We studied the potential of fosmidomycin, an antimalarial drug blocking the MEP pathway, to inhibit E. tenella and T. gondii growth in vitro. The drug was poorly effective even at high concentrations. Thus, both fosmidomycin sensitivity and isoprenoid metabolism differs substantially between apicomplexan species. PMID- 17399706 TI - Trypanosoma cruzi: populations bearing opposite virulence induce differential expansion of circulating CD3+CD4-CD8- T cells and cytokine serum levels in young and adult rats. AB - The JG strain is the least virulent while the CL-Brener clone is one of the most virulent Trypanosoma cruzi populations in young rats. In this study, we determined that the parasitemia peak values in CL-Brener clone-infected adult rats were 50-fold lower than in young rats and that mortality was null as compared to 45% death in young rats. Low parasitemia, milder and sustained myocarditis and myositis characterized JG infections. CL-Brener clone caused a significantly higher production of pro-inflammatory cytokines and higher expansion of CD3(+)CD4(-)CD8(-), double-negative (DN) T cells, during the acute phase in both adult and young rats. DN T cell frequencies correlated with IFN gamma levels. These findings may explain the higher inflammation and fast acute phase resolution in CL-Brener infection. In young rats, IL-10 levels were similar in both infections. The IL-10/IFN-gamma ratio was higher in JG acute infection in accordance with the milder inflammation and parasite persistence leading to a chronic phase. In conclusion, virulence and pathogenicity depend on T. cruzi ability to induce expansion of DN T cells and production of specific cytokines. PMID- 17399707 TI - DNA-probes for the highly sensitive identification of single nucleotide polymorphism using single-molecule spectroscopy. AB - This article presents a new, highly sensitive method for the identification of single nucleotide polymorphisms (SNPs) in homogeneous solutions using fluorescently labeled hairpin-structured oligonucleotides (smart probes) and fluorescence single-molecule spectroscopy. While the hairpin probe is closed, fluorescence intensity is quenched due to close contact between the chromophore and several guanosine residues. Upon hybridization to the respective target SNP sequence, contact is lost and the fluorescence intensity increases significantly. High specificity is achieved by blocking sequences containing mismatch with unlabeled oligonucleotides. Time-resolved single-molecule fluorescence spectroscopy enables the detection of individual smart probes passing a small detection volume. This method leads to a subnanomolar sensitivity for this single nucleotide specific DNA assay technique. PMID- 17399708 TI - Synergistic action of polyanionic and non-polar cofactors in fibrillation of human islet amyloid polypeptide. AB - Recent studies have led us to suppose that synergistic action of multiple solute cofactors could play substantial roles in amyloid-type fibrillogenesis of pathogenic polypeptides. To support this view, we performed aggregation experiments of human islet amyloid polypeptide (IAPP) in media containing both polyanions and non-polar solvents. The results demonstrated that the fibrillation at sub-micromolar IAPP occurred only when polyanionic and non-polar solutes coexist. A simple sum of two independent cofactor's effects could not account for the synergistic action. We propose that this synergy of polyanionic and nonpolar milieus could substantially modify the amyloidgenesis in the human body. PMID- 17399709 TI - Intermonomer electron transfer in the bc1 complex dimer is controlled by the energized state and by impaired electron transfer between low and high potential hemes. AB - The cytochrome bc(1) complex (commonly called Complex III) is the central enzyme of respiratory and photosynthetic electron transfer chains. X-ray structures have revealed the bc(1) complex to be a dimer, and show that the distance between low potential (b(L)) and high potential (b(H)) hemes, is similar to the distance between low potential hemes in different monomers. This suggests that electron transfer between monomers should occur at the level of the b(L) hemes. Here, we show that although the rate constant for b(L)-->b(L) electron transfer is substantial, it is slow compared to the forward rate from b(L) to b(H), and the intermonomer transfer only occurs after equilibration within the first monomer. The effective rate of intermonomer transfer is about 2-orders of magnitude slower than the direct intermonomer electron transfer. PMID- 17399710 TI - Sequence effects of single base loops in intramolecular quadruplex DNA. AB - We have examined the properties of intramolecular G-quadruplexes in which the G3 tracts are separated by single base loops. The most stable complex contained 1',2'-dideoxyribose in all three loops, while loops containing T and C were slightly less stable (by about 2 degrees C). Quadruplexes containing loops with single A residues were less stable by 8 degrees C for each T to A substitution. These folded sequences display similar CD spectra, which are consistent with the formation of parallel stranded complexes with double-chain reversal loops. These results demonstrate that loop sequence, and not just length, affects quadruplex stability. PMID- 17399711 TI - Spatial and temporal distribution of the ecdysteroid receptor (EcR) in haemocytes and epidermal cells during wound healing in the crayfish, Procambarus clarkii. AB - Wound healing in crustaceans preserves the integrity of the integument and prevents entry of pathogens. We studied the interaction between the moulting hormones (ecdysteroids) and the cellular events under the wound during wound healing with or without bacteria infection. Wounding of the carapace by abrasion induced a rapid increase in circulating ecdysteroid levels to a low sustained plateau level for about 12 days, followed by a sharp premoult peak and moulting. Within 48h of wounding, the nuclear receptor for ecdysteroids (EcR) appeared in the nuclei of haemocytes (hyaline, semigranular and granulocytes), visualized by confocal laser scanning microscopy and anti-EcR. Hyaline haematocytes aggregated in layers below the wound site and granulocytes engaged in phagocytosis. Therefore, the immune system responds directly and rapidly to ecdysteroids. Epidermal cells developed EcR only several days after the haemocytes and only under intact carapace, not under the wound where they appeared apoptotic. At the wound margin, EcR-positive epidermal cells and fibroblasts proceeded to migrate across the wound between the layers of haemocytes. Epidermis was fully regenerated by day 15; at this time the ecdysteroid titre began rising towards a premoult peak and EcR disappeared from the nuclei of epidermal cells suggesting that high amounts of ecdysteroids exert negative control on EcR. When bacteria were injected at the time of wounding, both the plateau level of ecdysteroid titre and the cellular events of wound healing were prolonged by 5-7 days, showing that healing of the wound is slower and that the duration of the plateau phase of the titre depends on the degree of assault on the animal. We conclude that the low levels of ecdysteroids induced by wounding activate the immune system to begin healing below the wound and also stimulate adjacent epidermal cells to commence the process of wound repair. PMID- 17399712 TI - Summary of controversial debates during the 5th "Homburg Pelvic Course" 13-15 September 2006. AB - Based on low incidence and lack of personal experience only few evidence based studies exist on several questions in pelvic and acetabular surgery. As part of an international consensus pelvic and acetabular course personal preferences and experience of an distinguished faculty and senior participants were discussed and we summarize in the paper the consented opinions and trends. Topics included the emergency treatment of life threatening pelvic ring injuries, treatment strategies in unstable sacral fractures, preferred surgical methods for transiliosacral screw fixation of the posterior pelvic ring, the value of CT and conventional radiographs in diagnostic of acetabular fractures, the choice of approach for treatment of acetabular fractures, the open vs. arthroscopic treatment of the femoro acetabular impingement of the hip and the treatment modalities in pelvic and acetabular fractures in geriatric patients. One has to keep in mind that this statements may help in the process of personal decision making in this difficult surgical field, but should not act as evidence based recommendations. PMID- 17399713 TI - Treatment options of pelvic and acetabular fractures in patients with osteoporotic bone. AB - The incidence of pelvic ring and acetabular fractures in the elderly is climbing relentlessly. This increase is attributed to a greater longevity and a decrease in the incidence of alcohol-related trauma in younger adults. Often, the elderly trauma patient has compromised physiological reserve and healing capacity due to concomitant morbidities, resulting in a less favourable clinical outcome. The presence of osteopenic or osteoporotic bone and other treatments for existing comorbidities hamper some treatment alternatives, especially those designed for younger patients. Diverse clinical presentations include minor trauma, major polytrauma and insufficiency fractures. An assessment of the general health and functional status of the patient is of utmost importance to determine the optimal treatment. The different treatment options of pelvic and acetabular fractures in the presence of osteoporosis vary mainly according to the clinical presentation and include: conservative methods, percutaneous or minimally invasive procedures, open reduction and fixation, and primary total hip arthroplasty. Whichever treatment is chosen, even for elderly people, the aim is a rapid mobilisation of the patient in order to reduce complications to some extent inherent to this age group. PMID- 17399715 TI - Effect of injection duration on bruising associated with subcutaneous heparin: a quasi-experimental within-subject design. AB - AIM: The purpose of this study was to determine and compare the effects of three different injection durations on bruising associated with subcutaneous heparin. BACKGROUND: Subcutaneous administration of the anticoagulant heparin sodium is a frequently performed nursing intervention. Heparin preparations are widely used for the post surgical prophylaxis, deep vein thrombosis and pulmonary emboli. Subcutaneous heparin injection often causes problems such as bruise, pain, induration, and hematoma at the injection site. Bruising that result from heparin injections may lead to anxiety, disruption of body image, the rejection of the treatment in patients, and the reduction of the reliance of patient to nurse. METHOD: The study used a quasi-experimental design. The population for the research was formed by the patients who were hospitalized at Chest Disease and Surgery Hospital and ordered to receive subcutaneous low-molecular weight heparin (LMWH). The research involved 36 chronic obstructive pulmonary disease (COPD) patients. Each subject received three injections by the same investigator using three different techniques. Site bruising was measured at 48h after each injection. The bruising size was measured using a plastic ruler. Descriptive statistics, Wilcoxon signed-rank and McNemar chi(2) tests were used to evaluate of the data. The level of significance was determined at P<0.05. RESULT: The 30s duration injection technique and waiting 10s before withdrawing the needle after 10s injection duration resulted in significantly smaller and less bruises than 10s duration injection technique. CONCLUSION: The 30s duration or waiting 10s before withdrawing the needle after 10s injection duration techniques should be used for subcutaneous heparin injections in clinical practice. PMID- 17399714 TI - The use of chlorhexidine to reduce maternal and neonatal mortality and morbidity in low-resource settings. AB - Of the 4 million neonatal deaths and 500,000 maternal deaths that occur annually worldwide, almost 99% are in developing countries and one-third are associated with infections. Implementation of proven interventions and targeted research on a select number of promising high-impact preventative and curative interventions are essential to achieve Millennium Development Goals for reduction of child and maternal mortality. Feasible, simple, low-cost interventions have the potential to significantly reduce the mortality and severe morbidity associated with infection in these settings. Studies of chlorhexidine in developing countries have focused on three primary uses: 1) intrapartum vaginal and neonatal wiping, 2) neonatal wiping alone, and 3) umbilical cord cleansing. A study of vaginal wiping and neonatal skin cleansing with chlorhexidine, conducted in Malawi in the 1990s suggested that chlorhexidine has potential to reduce neonatal infectious morbidity and mortality. A recent trial of cord cleansing conducted in Nepal also demonstrated benefit. Although studies have shown promise, widespread acceptance and implementation of chlorhexidine use has not yet occurred. This paper is derived in part from data presented at a conference on the use of chlorhexidine in developing countries and reviews the available evidence related to chlorhexidine use to reduce mortality and severe morbidity due to infections in mothers and neonates in low-resource settings. It also summarizes issues related to programmatic implementation. PMID- 17399716 TI - Dose-dependent coronary artery intimal thickening after local delivery of the anti-oxidant tetradecylthioacetic acid from stents. AB - OBJECTIVE: To examine the in vitro uptake and elution of the anti-oxidant tetradecylthioacetic acid (TTA) from phosphorylcholine (PC)-coated stents, and the in vivo uptake, retention, inflammatory response and histomorphometric changes after overstretch injury of the porcine coronary artery. METHODS: PC coated stents were loaded in one of three different concentrations of TTA (87, 174 and 347 mmol/L, i.e. 25, 50 and 100 mg/mL) and randomized versus PC-coated stents to the right coronary or left circumflex artery (18 pigs). Uptake of TTA into the coronary wall from the 347 mmol/L concentration was measured after 3 h and 24 h, 7 days, 14 days and 28 days (two pigs at each time point). RESULTS: In vitro, TTA was successfully loaded onto the stents and elution was nearly complete after 48 h. In vivo, TTA could be demonstrated in the vessel wall for up to 4 weeks. Percent area stenosis was significantly higher in the TTA group, 35.2+/-20.9% versus 27.5+/-17.0% (p=0.03). Dose-related comparison showed increased intimal thickness, 0.66+/-0.53 mm versus 0.29+/-0.26 mm (p=0.008) and intimal area, 2.83+/-1.61 mm2 versus 1.58+/-0.91 mm2 (p=0.004) for the 347 mmol/L TTA versus controls. There was a significantly positive relationship between the TTA-loading dose and both intimal area (B=0.69, p=0.01) and maximal intimal thickness (B=0.17, p=0.02). The pro-inflammatory precursor arachidonic acid increased four-fold in the arterial wall of the TTA group, while the anti inflammatory fatty acid index, calculated as (docosapentaenoic acid+docosahexaenoic acid+dihomo-linolenic acid)/arachidonic acid, was suppressed to 0.65+/-0.27 compared to 1.13+/-0.23 in control vessels (p<0.001). CONCLUSION: TTA caused a dose-dependent intimal thickening and reduced anti-inflammatory fatty acid index. Contrary to expectations, TTA seems unsuitable as stent coating. PMID- 17399717 TI - Decreased post-prandial triglyceride response and diminished remnant lipoprotein formation in cholesteryl ester transfer protein (CETP) deficiency. AB - Plasma cholesteryl ester transfer protein (CETP) mediates CE/TG exchange among various lipoproteins. CETP deficiency results in low LDL and high HDL phenotype including apoE-rich large HDL. Large HDL could provide apoE to chylomicron/VLDL during lipolysis in post-prandial state, accelerating remnant lipoprotein uptake in the liver. To determine the effects of low CETP levels on post-prandial lipoprotein metabolism, lipid levels of plasma remnant-like lipoprotein particles (RLP) fraction were determined in one homozygous and three heterozygous CETP deficiency and controls with apoE3/3 phenotype. After oral fat-load, the area under curve (AUC) of TG levels were remarkably decreased in CETP deficiency as compared to controls (423+/-187 [S.D.] mg/dl x h in three heterozygous CETP deficiency and 926+/-268 [S.D.] in 10 controls, P=0.012). Similarly, the homozygote had a low AUC of TG levels (416 mg/dl x h). Plasma RLP-cholesterol levels were decreased in heterozygotes, but not significantly as compared to controls (P=0.14). HPLC analysis showed that increased RLP-cholesterol level was not due to conventional VLDL-LDL size RLP, but to those in large HDL size in the homozygote. In heterozygotes, bimodal distribution of RLP-cholesterol level was found in lipoprotein sizes of conventional VLDL-LDL and large HDL. Subjects with CETP deficiency appeared to have low levels of TG response and diminished remnant lipoprotein formation after fat-load. PMID- 17399719 TI - No evidence for modulation of endothelial nitric oxide synthase by the olive oil polyphenol hydroxytyrosol in human endothelial cells. AB - Reduced nitric oxide (NO) availability is associated with the development of atherosclerosis. Upregulation of endothelial nitric oxide synthase (eNOS) activity is pursued as a strategy for the prevention of cardiovascular diseases. The polyphenol hydroxytyrosol (HT) which is present in olive oil and red wine, is regarded to be partly responsible for the beneficial effects associated with olive oil consumption and has shown antiatherogenic activity in vitro and in vivo. To elucidate the underlying molecular mechanisms, we investigated possible effects of HT on the endothelial nitric oxide synthase (eNOS). We used human endothelial cells (EA.hy926) and examined eNOS on three different levels, addressing eNOS promoter transactivation, eNOS enzyme activity and nitric oxide availability. Cells were treated with a broad range of HT concentrations (from 10 nM to 100 microM) and for different incubation times (15 min to 24 h). HT did not exert significant positive effects on eNOS in any of our assay systems. Neither did we find evidence for a possible synergism between the red wine polyphenol resveratrol and HT. We conclude that a direct modulation of eNOS is unlikely to account for the antiatherogenic properties of HT under non-inflammatory conditions. PMID- 17399720 TI - Two new large deletions in the low density lipoprotein receptor (LDLR) gene not revealed by PCR-based molecular diagnosis of familial hypercholesterolemia. AB - In the French Canadian population six mutations appear to be responsible for about 85% of FH cases. Two of these mutations are large deletions. The most prevalent deletion is a >15 kb deletion of the promoter and first exon; the second, a 5 kb deletion that removes exons 2 and 3. The high frequency of these deletions in the French Canadian population has been attributed to a founder effect. Other mutations are present in the population but at a much lower prevalence. We recently identified two new large deletions in FH patients of French Canadian descent. Carriers of the new deletions were identified because of an unusual pattern of band migration on Southern blots. We have identified and sequenced the deletions' boundaries. The first deletion covers 3813 bp and removes exons 7 and 8. The second deletion covers 5994 bp and removes exons 3-6. These deletions have not been previously reported. They would have been missed if a PCR-based method had been used instead of Southern blot analysis. PMID- 17399721 TI - Biomechanical response of the pubic symphysis in lateral pelvic impacts: a finite element study. AB - Automotive side impacts are a leading cause of injuries to the pubic symphysis, yet the mechanisms of those injuries have not been clearly established. Previous mechanical testing of isolated symphyses revealed increased joint laxity following drop tower lateral impacts to isolated pelvic bone structures, which suggested that the joints were damaged by excessive stresses and/or deformations during the impact tests. In the present study, a finite element (FE) model of a female pelvis including a previously validated symphysis sub-model was developed from computed tomography data. The full pelvis model was validated against measured force-time impact responses from drop tower experiments and then used to study the biomechanical response of the symphysis during the experimental impacts. The FE models predicted that the joint underwent a combination of lateral compression, posterior bending, anterior/posterior and superior/inferior shear that exceeded normal physiological levels prior to the onset of bony fractures. Large strains occurred concurrently within the pubic ligaments. Removal of the contralateral constraints to better approximate the boundary conditions of a seated motor vehicle occupant reduced cortical stresses and deformations of the pubic symphysis; however, ligament strains, compressive and shear stresses in the interpubic disc, as well as posterior bending of the joint structure remained as potential sources of joint damage during automotive side impacts. PMID- 17399722 TI - Biomechanical response of collagen fascicles to restressing after stress deprivation during culture. AB - In vitro tissue culture experiments were performed to study the biomechanical response of collagen fascicles to restressing after exposure to non-loaded condition. Collagen fascicles of approximately 300 microm in diameter were aseptically dissected from rabbit patellar tendons. They were cultured under no load condition for 1 week, and then under a static stress of approximately 1.2 MPa for the subsequent 1 or 2 weeks. After culture, their mechanical properties were determined with a micro-tensile tester, and were compared to those of fascicles cultured under no-load condition and non-cultured, control fascicles. Tangent modulus and tensile strength of the non-loaded fascicles were significantly lower than those of the control fascicles at 1 week and gradually decreased thereafter. However, the modulus and strength were increased by restressing. After 2-week restressing, both parameters were significantly greater than those of the time-matched, non-loaded fascicles, although these values were still significantly lower than those of the control fascicles. That is, the application of stress after exposure to non-loaded condition suppressed the deterioration of the biomechanical properties of fascicles, although it did not improve. These results indicate that a short period of stressing is not sufficient for cultured collagen fascicles to completely recover their mechanical properties, if they are once exposed to no-stress condition even for a short period of time. These are similar to previous results observed in tendons and ligaments inside the body. PMID- 17399723 TI - In vitro confocal micro-PIV measurements of blood flow in a square microchannel: the effect of the haematocrit on instantaneous velocity profiles. AB - A confocal microparticle image velocimetry (micro-PIV) system was used to obtain detailed information on the velocity profiles for the flow of pure water (PW) and in vitro blood (haematocrit up to 17%) in a 100-microm-square microchannel. All the measurements were made in the middle plane of the microchannel at a constant flow rate and low Reynolds number (Re=0.025). The averaged ensemble velocity profiles were found to be markedly parabolic for all the working fluids studied. When comparing the instantaneous velocity profiles of the three fluids, our results indicated that the profile shape depended on the haematocrit. Our confocal micro-PIV measurements demonstrate that the root mean square (RMS) values increase with the haematocrit implying that it is important to consider the information provided by the instantaneous velocity fields, even at low Re. The present study also examines the potential effect of the RBCs on the accuracy of the instantaneous velocity measurements. PMID- 17399724 TI - Hysteretic pinching of human secondary osteons subjected to torsion. AB - The mechanical behavior of bone tissue's ultra- and micro- structure is fundamental to assessment of macroscopic bone mechanics. This paper explores the ultra-structural characteristics of human femoral tissue responsible for energy absorption of secondary osteons under mechanical loading. A novel mathematical interpretation of single osteon mechanics elucidates the behavior of the collagen apatite interface. Fully calcified single osteon specimens were mechanically tested quasi-statically under cyclic torsional loading about their longitudinal axis. On each hysteretic diagram, all cycles after the initial monotonic cycle appear pinched and share two points. Stiffness degradation and pinching degradation were investigated on the torque versus deflection-angle-per-unit length diagrams as the number of cycles increases, in relation to the appearance of osteons in cross-section under circularly polarized light microscopy. Material science's Bauschinger effect, originally defined for metals and later extended to structures reinforced with metal bars, is adapted to describe pinching. Material science's prying effect, defined as amplification of eccentric tensile load through lever action, is employed to explain pinching. The presence of the two points shared by all complete cycles is analyzed in terms of the mathematical fixed point theorem. The results allow formulation of the following conjectures: (1) the prying of carbonated apatite crystallites at the interface with the 40 nm long bands of non-calcified collagen fibrils causes pinching; (2) the prying effect increases with the increasing percentage of collagen-apatite elements that form a larger angle with the osteon axis; and (3) micro-cracks increase more in number than in length as the number of cycles increases. PMID- 17399725 TI - Acceleration slope of exercise-induced impacts is a determinant of changes in bone density. AB - High acceleration levels (>4g) seen during impact exercises have been shown to increase bone mineral density (BMD) in premenopausal women. The aim of this study was to examine how the other acceleration signal characteristics, i.e. the slope, area and energy of the signal are related to changes in bone density, using long term quantification of physical activity. Daily physical activity was continuously assessed with a waist-worn accelerometer-based body movement monitor in 64 premenopausal women participating in a 12-month population-based exercise trial. The daily number of exercise-induced impacts at different slope, area and energy levels of the acceleration signal was analyzed. Physical activity inducing slopes 1000 m/s(3), acceleration peak areas 2m/s or signal energies 75 m(2)/s(3) was associated with BMD change in the hip (p<0.05). Impacts with the smallest slopes (<1000 m/s(3)) were positively associated with changes in calcaneal speed of ultrasound, while impacts with slopes 1500 m/s(3) or areas 4m/s were positively correlated with broadband ultrasound attenuation changes (p<0.05). We conclude that the acceleration slope of exercise-induced impacts is an important determinant of bone density. The slope threshold for improving BMD at the hip is 1000 m/s(3), which can be achieved during normal exercise including fast movements such as running and jumping. PMID- 17399726 TI - Simultaneous determination of tyramine and tryptamine and their precursor amino acids by micellar liquid chromatography and pulsed amperometric detection in wines. AB - Two biogenic amines, tryptamine and tyramine, and their precursors, tryptophan and tyrosine, were determined by a liquid chromatographic procedure. A hybrid micellar mobile phase of sodium dodecyl sulphate (SDS) and 1-propanol, a C18 column and electrochemical detection were used. A pH study in the range of 3-9 was performed and pH 3 was finally selected in accordance with resolution and analysis time. Oxidation potential was also checked in the range 0.6-0.9V: the maximum area obtained in all those potentials was at 0.8V, which was selected to carry out the analysis using a sequence of pulsed amperometric detection waveform. The four compounds were resolved using a mobile phase of 0.15M SDS-5% 1 propanol with an analysis time of 16 min. Repeatabilities and intermediate precision were evaluated at three different concentrations for each compound with RSD values lower than 2.6 and 4.8%, respectively. Limits of detection and quantification were also obtained within the 10-40 and 33-135 ng/ml ranges, respectively. Finally, the applicability of the procedure was tested in several types of wine and no matrix effect was observed. The possibility of direct sample introduction simplifies and greatly expedites the treatments with reduced cost, improving the accuracy of the procedures. PMID- 17399727 TI - Determination of pesticides and metabolites in wine by high performance liquid chromatography and second-order calibration methods. AB - The models parallel factor analysis (PARAFAC) and the recently introduced bilinear least squares (BLLS) were applied to develop second-order calibration methods to high performance liquid chromatography with diode array detection (HPLC-DAD) data, where overlap of interferences with the compounds of interest was observed, making the determination and resolution of the analytes possible. In this work, the simultaneous determination of five pesticides and two metabolites in wine samples by HPLC-DAD was performed, using the second-order advantage. The results of two chromatographic methods were compared, involving either isocratic or gradient elution. An appropriate preprocessing method was necessary to correct the effects of time shifts, baseline variations and background. BLLS presented results that were of the same quality as PARAFAC in five cases, but in two other situations only PARAFAC enabled analyte quantitation. Relative errors of prediction lower than 10% for all compounds were obtained, indicating that the methodology employing HLPC-DAD and second-order calibration can handle complex analytical systems. PMID- 17399728 TI - Analysis of polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls in vegetable oil samples by gas chromatography-ion trap tandem mass spectrometry. AB - Gas chromatography coupled to ion trap tandem mass spectrometry (CG-MS-MS) has been evaluated for the analysis of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in vegetable oil samples containing different concentration levels (0.2-6 pg WHO-TEQ g(-1) for both PCDD/Fs and dl-PCBs) of the 29 toxic congeners of PCDD/F and dioxin-like PCBs. The effect of potential interfering compounds such as polychlorinated naphthalenes (PCNs), polychlorinated biphenyls (PCBs) and polychlorinated diphenylethers (PCDEs) on the quantification of mono-ortho PCBs has been investigated. In addition, the influence of the clean-up procedure on the final determination by CG-MS-MS was studied, showing that the quality of the results depends to a great extent on this analytical step. Quality parameters have been established and good precisions (CV: 3-19%) and low limits of detection for PCDD/Fs (0.04-0.20 pg g(-1) oil) and dl-PCBs (0.08-0.64 pg g(-1) oil) were obtained. The method was validated by a comparison of the CG-MS-MS results with those obtained by GC-HRMS. PMID- 17399729 TI - Comprehensive separation of secondary metabolites in natural products by high speed counter-current chromatography using a three-phase solvent system. AB - High-speed counter-current chromatography (HSCCC) using the three-phase solvent system n-hexane-methyl acetate-acetonitrile-water at a volume ratio of 4:4:3:4 was applied to the comprehensive separation of secondary metabolites in several natural product extracts. A wide variety of secondary metabolites in each natural product was effectively extracted with the three-phase solvent system, and the filtered extract was directly submitted to the HSCCC separation using the same three-phase system. In the HSCCC profiles of crude natural drugs listed in the Japanese Pharmacopoeia, several physiologically active compounds were clearly separated from other components in the extracts. The HSCCC profiles of several tea products, each manufactured by a different process, clearly showed their compositional difference in main compounds such as catechins, caffeine, and pigments. These HSCCC profiles also provide useful information about hydrophobic diversity of whole components present in each natural product. PMID- 17399730 TI - Analysis of triacylglycerol and fatty acid isomers by low-temperature silver-ion high performance liquid chromatography with acetonitrile in hexane as solvent: limitations of the methodology. AB - Silver ion HPLC (Ag-HPLC), utilizing columns containing silver ions bonded to a silica substrate and acetonitrile in hexane as solvent, has proven to be a powerful technology for the analysis of geometric (cis or trans) or positional fatty acids, fatty acid ester (primarily methyl ester; FAME), or triacylglycerol (TAG) isomers. Previous studies had demonstrated that, unlike gas chromatography, samples eluted more rapidly at lower temperatures (at 20 degrees C versus 40 degrees C, for example). A low-temperature bath [dual-column Ag-HPLC; isocratic solvent systems of 0.3 to 0.7% acetonitrile (ACN) in hexane] was utilized to study the application of this system at low (below 0 degrees C) temperatures for analysis of FAME (zero to six double bonds) and TAG [SSS, OOO and LLL, where S=stearic acid (18:0), O=oleic acid (9c-18:1), and L=linoleic acid (9c, 12c 18:2)] standards. While FAME elution times continued to decrease from 0 degrees C to -10 degrees C, they began to increase at -20 degrees C. A similar situation was noted for the TAG isomers, except that retention times began to increase below 0 degrees C. The lower temperature limit of the Ag-HPLC/ACN in hexane system is thus ca. -25 degrees C. Increasing sample elution times and pump head pressures upon sample injection were noted at temperatures of -25 degrees C to 40 degrees C. Equilibration times at each temperature could be reduced to ca. 15 min without loss of resolution and with retention times of +/-2%. Temperature, rather than solvent composition, can therefore be utilized with the Ag-HPLC/ACN in hexane solvent system to optimize elution times and resolution(s) of FAME and TAG isomers. PMID- 17399731 TI - Large volume injection in ion chromatography Separation of rubidium and strontium for on-line inductively coupled plasma mass spectrometry determination of strontium isotope ratios. AB - Large volume injection, up to 5 mL, was evaluated and optimised for the on-line ion chromatographic separation of Rb and Sr before ICP-MS measurement of Sr isotope ratios. Flat-topped chromatographic peaks, ideally suited for multicollector ICP-MS isotope ratio measurements, could be obtained when the composition of the mobile phase (nitric acid and 18-crown-6 ether) was identical to the matrix of the sample. Under those conditions rubidium eluted at the dead volume of the column while strontium produced a flat-topped transient signal with several minutes of stable plateau. On-line data acquisition during several minutes at the plateau of Sr signal allowed high precision Sr isotope ratio measurement. The developed procedure was evaluated for Sr isotope ratio measurements on different types of samples, including cider, apples, apple leaves, and soil extracts, in the frame of a long-term project aiming at origin authentication using strontium isotope ratio measurements. It was observed that sample matrix caused broadening of the strontium chromatographic peak and loss of flat-topped peak profile. Under those circumstances the addition of the complexing crown-ether 18-crown-6 both to samples and chromatographic eluent provided two distinct advantages. First, a drastic increase in the retention of strontium was observed which could be modulated by increasing the concentration of nitric acid in the eluent up to 900 mM. This increase in the eluent HNO(3) concentration allowed the application of the method to acid soil digests and other high acidity samples. Second, the matrix of the sample did not affect any more the chromatographic peak profile and similar chromatographic separations could be obtained for samples and standards maintaining the flat-topped Sr peak profile. Sample preparation consisted of a simple 1:10 dilution of the cider or pre-treated solid samples by adding HNO(3) (900 mM) and 18-crown-6 ether (5mM) to obtain similar composition in the sample solution and the HPLC eluent. PMID- 17399732 TI - Microwave-accelerated derivatization for the simultaneous gas chromatographic mass spectrometric analysis of natural and synthetic estrogenic steroids. AB - A rapid microwave-accelerated derivatization process for the GC-MS analysis of steroid estrogens, estrone (E1), 17beta-estradiol (E2), estriol (E3), 17alpha ethynylestradiol (EE2) and mestranol (MeEE2), was developed. Under microwave irradiation, the five estrogenic hormones studied were simultaneously derivatized with N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA)+trimethylchlorosilane (TMCS) in pyridine solution. Effects of irradiation time (15-120 s) and power level (240-800 W) on the yield of the derivatization were investigated. The derivatization under the irradiation of 800 W microwave for 60s produced comparable results when compared with the conventional heating process in a sand bath for 30 min at 80 degrees C in terms of derivatization yield, linearity and precision for all steroid hormones tested. The calibration curves are linear between 3.00 and 3.00 x 10(2) microg mL(-1). The square of the regression coefficients (R(2)) range from 0.979 to 1.000. The applicability of the method was evaluated on spiked river and distilled water samples at two concentrations, 25.0 and 2.00 x 10(2) ng mL(-1). The recoveries obtained by using microwave heating (60s, 800 W) were similar to those by conventional heating. When combined solid-phase extraction (SPE) with the application of the microwave-accelerated derivatization proposed here, the detection limits of 0.02-0.1 ng L(-1) for the steroid hormones have been achieved. The results demonstrated that microwave accelerated derivatization is an efficient and suitable sample preparation method for the GC-MS analysis of estrogenic steroids. PMID- 17399733 TI - Application of a chromatography model with linear gradient elution experimental data to the rapid scale-up in ion-exchange process chromatography of proteins. AB - We applied the model described in our previous paper to the rapid scale-up in the ion exchange chromatography of proteins, in which linear flow velocity, column length and gradient slope were changed. We carried out linear gradient elution experiments, and obtained data for the peak salt concentration and peak width. From these data, the plate height (HETP) was calculated as a function of the mobile phase velocity and iso-resolution curve (the separation time and elution volume relationship for the same resolution) was calculated. The scale-up chromatography conditions were determined by the iso-resolution curve. The scale up of the linear gradient elution from 5 to 100mL and 2.5L column sizes was performed both by the separation of beta-lactoglobulin A and beta-lactoglobulin B with anion-exchange chromatography and by the purification of a recombinant protein with cation-exchange chromatography. Resolution, recovery and purity were examined in order to verify the proposed method. PMID- 17399734 TI - Canine subcutaneous mast cell tumour: diagnosis and prognosis. AB - The aim of this study was to characterize the pathology and clinical outcome of the subcutaneous variant of canine mast cell tumour. Fifty-three cases satisfying the inclusion criteria were selected from the pathology archive of the College of Veterinary Medicine, University of Tennessee. Referring veterinarians provided information on outcome. These dogs had a median age of 9 years (range 3-17 years). After characterizing tumours histologically, nuclear expression of proliferating cell nuclear antigen (PCNA) and Ki67 (MIB-1 clone) was determined immunohistochemically and mast cell origin was confirmed with c-Kit staining. Counts of argyrophilic nucleolar organizer regions (AgNOR) were determined by silver staining. Nuclear labelling was counted in 100 tumour cells. Margins were recorded as incomplete in 66% of dogs, and metastases occurred in 6% of dogs. The estimated minimum mean survival time from date of diagnosis was 1199 days, ranging from 55 to >1780 days. The median scores from immunohistochemical labelling were PCNA 0.05 and Ki67 0.03 per 100 tumour cells. The median score for AgNOR staining was 1.25 per 100 tumour cells. The patterns of c-Kit expression included membranous labelling in 20 tumours, stippled cytoplasmic labelling in 23 tumours and diffuse cytoplasmic labelling in 10 tumours. Age (r=-0.61, P=0.14) and AgNOR score (r=-0.58, P=0.17) had moderate, but non-significant, negative associations with survival. PCNA (r=-0.32, P=0.47), Ki67 (r=-0.22, P=0.64) and c Kit immunolabelling was not associated with survival. The subcutaneous variant of canine mast cell tumour is distinct in having features of intermediate histological grade and extended mean survival times, suggesting a slightly better long-term prognosis than for higher grade dermal variants. Expression of nuclear proliferation markers is not associated with outcome. PMID- 17399735 TI - Automatic activation of phonology in silent reading is parallel: evidence from beginning and skilled readers. AB - The picture-word interference paradigm was used to shed new light on the debate concerning slow serial versus fast parallel activation of phonology in silent reading. Prereaders, beginning readers (Grades 1-4), and adults named pictures that had words printed on them. Words and pictures shared phonology either at the beginnings of words (e.g., DOLL-DOG) or at the ends of words (e.g., FOG-DOG). The results showed that phonological overlap between primes and targets facilitated picture naming. This facilitatory effect was present even in beginning readers. More important, from Grade 1 onward, end-related facilitation always was as strong as beginning-related facilitation. This result suggests that, from the beginning of reading, the implicit and automatic activation of phonological codes during silent reading is not serial but rather parallel. PMID- 17399736 TI - Morphological and molecular characteristics of a new species of Pasteuria parasitic on Meloidogyne ardenensis. AB - A species of the hyper-parasitic bacterium Pasteuria was isolated from the root knot nematode Meloidogyne ardenensis infecting the roots of ash (Fraxinus excelsior). It is morphologically different from some other Pasteuria pathogens of nematodes in that the spores lack a basal ring on the ventral side of the spore and have a unique clumping nature. Transmission electron microscopy (TEM) showed that the clumps of spores are not random aggregates but result from the disintegration of the suicide cells of the thalli. Sporulation within each vegetative mycelium was shown to be asynchronous. In addition to the novel morphological features 16S rRNA sequence analysis showed this to be a new species of Pasteuria which we have called P. hartismeri. Spores of P. hartismeri attach to juveniles of root-knot nematodes infecting a wide range of plants such as mint (Meloidogyne hapla), rye grass (unidentified Meloidogyne sp.) and potato (Meloidogyne fallax). PMID- 17399737 TI - Insecticidal bacteria isolated from predatory larvae of the antlion species Myrmeleon bore (Neuroptera: Myrmeleontidae). AB - Various bacterial species were isolated from the crop (digestive organ) of the antlion species Myrmeleon bore and tested for their insecticidal activity against caterpillars by injection. Sixty-eight isolates from the antlion crop were grouped into twenty-four species based on homologies of 16S rRNA gene sequences and biochemical properties. Isolated Bacillus cereus, Bacillus sphaericus, Morganella morganii, Serratia marcescens and a Klebsiella species killed 80% or more cutworms when injected at a dose of 5x10(5)cells per insect. In addition, cutworms killed by these isolates resembled observations made of caterpillars attacked by antlions. A culture-independent analysis showed that the isolated bacterial species are likely to be frequently present in the antlion crop. These results suggest that insecticidal microorganisms associate with antlions, and may promote the death of prey. PMID- 17399738 TI - Quantitative analysis of peroxisomal targeting signal type-1 binding to wild-type and pathogenic mutants of Pex5p supports an affinity threshold for peroxisomal protein targeting. AB - Peroxisomal biogenesis disorders (PBDs) are caused by mutations in 12 distinct genes that encode the components of the peroxisome assembly machinery. Three mutations in the gene encoding Pex5p, the peroxisomal targeting signal type-1 (PTS1) receptor, have been reported, each associated with a disorder of the Zellweger spectrum of different severity. Here, we report studies of the affinities of mutated forms of Pex5p for a series of PTS1 peptides and conclude that PTS1-affinity reductions are correlated with disease severity and cell biological phenotype. A quantitative model has been developed that allows estimation of the dissociation constants for complexes with a wide range of PTS1 sequences bound to wild-type and mutant Pex5p. In the context of this model, the binding measurements suggest that no PTS1-containing proteins are targeted by Pex5p(N489K) and only a relatively small subset of PTS1-containing proteins with the highest affinity for Pex5p are targeted to peroxisomes by Pex5p(S563W). Furthermore, the results of the analysis are consistent with an approximate dissociation constant threshold near 500 nM required for efficient protein targeting to peroxisomes. PMID- 17399739 TI - Crystal structure of the zinc-binding transport protein ZnuA from Escherichia coli reveals an unexpected variation in metal coordination. AB - Bacterial ATP-binding cassette transport systems for high-affinity uptake of zinc and manganese use a cluster 9 solute-binding protein. Structures of four cluster 9 transport proteins have been determined previously. However, the structural determinants for discrimination between zinc and manganese remain under discussion. To further investigate the variability of metal binding sites in bacterial transporters, we have determined the structure of the zinc-bound transport protein ZnuA from Escherichia coli to 1.75 A resolution. The overall structure of ZnuA is similar to other solute-binding transporters. A scaffolding alpha-helix forms the backbone for two structurally related globular domains. The metal-binding site is located at the domain interface. The bound zinc ion is coordinated by three histidine residues (His78, His161 and His225) and one glutamate residue (Glu77). The functional role of Glu77 for metal binding is unexpected, because this residue is not conserved in previously determined structures of zinc and manganese-specific transport proteins. The observed metal coordination by four protein residues differs significantly from the zinc-binding site in the ZnuA transporter from Synechocystis 6803, which binds zinc via three histidine residues. In addition, the E. coli ZnuA structure reveals the presence of a disulfide bond in the C-terminal globular domain that is not present in previously determined cluster 9 transport protein structures. PMID- 17399740 TI - Multivariate analysis of conserved sequence-structure relationships in kinesins: coupling of the active site and a tubulin-binding sub-domain. AB - An extensive computational analysis of available sequence and crystal structure data was used to identify functionally important residue interactions within the motor domain of the kinesin molecular motor. Principal component analysis revealed that all current kinesin crystal structures reside in one of two main conformations, which differ at the active site, and in the position of a microtubule-binding sub-domain relative to a rigid central core. This sub-domain consists of secondary structure elements alpha4-loop12-alpha5-loop13 and contains a conserved hydrophilic surface patch that may be involved in strong binding to microtubules. A hinge point for the sub-domain motion lies near a conserved glycine at position 292. Statistical coupling analysis revealed a network of co evolving positions that link this region to the nucleotide-binding site, via a highly conserved histidine in the switch I loop. The data are consistent with a model in which the nucleotide status of the active site shifts kinesin between weak and strong binding conformations via reconfiguration of the identified sub domain. Our data provide a statistically supported framework for further examination of this and other structure-function relationships in the kinesin family. PMID- 17399741 TI - The critical role of a hydrogen bond between Gln63 and Trp104 in the blue-light sensing BLUF domain that controls AppA activity. AB - AppA is a novel blue-light receptor that controls photosynthetic gene expression in the purple bacterium Rhodobacter sphaeroides. The photocycle reaction of the light-sensing domain, BLUF, is unique in the sense that a few hydrogen bond rearrangements are accompanied by only slight structural changes of the bound chromophore. However, the exact features of the hydrogen bond network around the active site are still the subject of some controversy. Here we present biochemical and genetic evidence showing that either Gln63 or Trp104 in the active site of the BLUF domain is crucial for light sensing, which in turn controls the antirepressor activity of AppA. Specifically, the Q63L and W104A mutants of AppA are insensitive to blue light in vivo and in vitro, and their activity is similar to that of the light-adapted wild-type AppA. Based on spectroscopic and structural information described previously, we conclude that light-dependent formation and breakage of the hydrogen bond between Gln63 and Trp104 are critical for the light-sensing mechanism of AppA. PMID- 17399742 TI - Peripheral neuropathy in patients with diabetic foot ulcers: clinical and nerve conduction study. AB - OBJECTIVES: Diabetic foot lesions develop predominantly in male patients and sensory neuropathy is the most frequent type of neuropathy associated with these lesions. The aim of this study was to analyze the clinical and electrophysiological features in a cohort of patients with diabetic foot. RESEARCH DESIGN AND METHODS: The recordings of 318 consecutive diabetic patients (127 women and 191 men) with an ongoing or healed foot ulcer who had been referred for electrophysiological consultation were evaluated retrospectively. RESULTS: 60.1% of our cohort were male. Loss of deep sensation and deep tendon reflex abnormalities were the most common neurological findings. Negative sensory symptoms (63.7% vs 40.8%, p<0.01) and neuropathic pain (38.5% vs 18.3%, p<0.01) were more frequent in females, whereas atrophy was more frequent in male patients (22.8% vs 46%, p<0.01). Motor nerve conduction abnormalities and ulnar nerve involvement was more frequent and severe in males. Abnormal electrophysiological findings were mild in 70 patients (female 42, 60%). In this group, hemiplegia, peripheral arterial disease, multiple bone fractures, end stage renal failure, recent pulmonary tuberculosis and dementia accompanied mild polyneuropathy. Thirty patients had shown prominent decrease in nerve conduction velocity which indicated severe demyelination. Among these 30 patients, 6 male subjects had clinical features similar to that of chronic inflammatory demyelinating polyneuropathy. CONCLUSIONS: Our results indicate that male gender, motor neuropathy and mononeuropathies, especially ulnar neuropathy is associated with the development of DF among our patients with DF. Patients with diabetes mellitus have a predisposition to develop chronic inflammatory demyelinating polyneuropathy and this may also facilitate formation of diabetic foot. History of hemiplegia, dementia and trauma are permissive risk factors for diabetic foot in the presence of mild polyneuropathy. PMID- 17399743 TI - Natural selection in chemical evolution. AB - We propose that chemical evolution can take place by natural selection if a geophysical process is capable of heterotrophic formation of liposomes that grow at some base rate, divide by external agitation, and are subject to stochastic chemical avalanches, in the absence of nucleotides or any monomers capable of modular heredity. We model this process using a simple hill-climbing algorithm, and an artificial chemistry that is unique in exhibiting conservation of mass and energy in an open thermodynamic system. Selection at the liposome level results in the stabilization of rarely occurring molecular autocatalysts that either catalyse or are consumed in reactions that confer liposome level fitness; typically they contribute in parallel to an increasingly conserved intermediary metabolism. Loss of competing autocatalysts can sometimes be adaptive. Steady state energy flux by the individual increases due to the energetic demands of growth, but also of memory, i.e. maintaining variations in the chemical network. Self-organizing principles such as those proposed by Kauffman, Fontana, and Morowitz have been hypothesized as an ordering principle in chemical evolution, rather than chemical evolution by natural selection. We reject those notions as either logically flawed or at best insufficient in the absence of natural selection. Finally, a finite population model without elitism shows the practical evolutionary constraints for achieving chemical evolution by natural selection in the lab. PMID- 17399744 TI - Post-ischaemic peak flow and myogenic flowmotion component are independent variables for skin post-ischaemic reactive hyperaemia in healthy subjects. AB - The aim of this study was to clarify whether the post-ischaemic amplification of skin blood flowmotion (SBF) influences the extent of skin post-ischaemic hyperaemia. Forearm skin perfusion was measured by means of laser Doppler flowmetry (LDF) and forearm SBF was examined using Fourier analysis of LDF signal, under basal conditions and following forearm ischaemia in 50 healthy subjects. Power spectral density (PSD) of SBF total spectrum (0.009-1.6 Hz), as well of the frequency intervals (FI) related to endothelial (0.009-0.02 Hz), sympathetic (0.02-0.06 Hz), myogenic (0.06-0.2 Hz), respiratory (0.2-0.6 Hz) and cardiac (0.6-1.6 Hz) activity was measured in PU(2) (LDF perfusion unit)/Hz. Multiple regression analysis evaluated whether post-ischaemic peak-flow, as an indicator of shear stress, or post-ischaemic SBF independently affected the post peak-flow hyperaemia calculated as corrected area under the LDF curve (C-AUC). Following ischaemia, we observed a statically significant increase in skin perfusion (from basal of 11.7+/-5.8 PU to peak flow of 62.3+/-41.4 PU, p<0.0000005) and in PSD of SBF total spectrum (p<0.01) as well of the different FI considered (p<0.005 for the endothelial and myogenic FI; p<0.05 for the sympathetic, respiratory and cardiac FI) compared to baseline. Multiple regression analysis showed that peak flow and post-ischaemic SBF component of myogenic origin were significant independent variables for the C-AUC (p=0.0000001 and p=0.009, respectively). These findings suggest that not only increased shear stress but also post-ischaemic amplification of myogenic SBF component independently contributes to the more prolonged phase of post-ischaemic skin re perfusion in healthy subjects. PMID- 17399745 TI - Action-perception dissociation; preserved reactive grip force despite tactile extinction due to cortical stroke. AB - Sensory extinction following stroke, manifests as a bias in spatial attention towards ipsilesional spatial locations, which arises when stimuli from other locations compete for pathologically limited attentional resources. In the tactile domain, extinction results in a failure to verbally report tactile stimuli applied to a contralesional body part when they are timed to coincide with ipsilesional tactile contact. While it is typical for research in this area to focus on the verbal report of sensory stimuli as a measure of conscious awareness, work in visual extinction has shown that when contralesional stimuli fail to reach conscious awareness, they may still contribute to the control of actions. We describe the case of a woman with tactile extinction who failed to verbally report contralesional tactile input associated with perturbations to bimanual grasp. Despite this, the same stimulus was sufficient to drive reflexive grip force responses. PMID- 17399746 TI - Alliesthesia in visual and auditory sensations from environmental signals. AB - 'Alliesthesia' describes the fact that sensory stimuli can arouse pleasant or unpleasant sensations according to the internal state of a person. In the present work, the hedonicity aroused by stimuli from the environment in visual and auditory sensations was evaluated in 5 situations: 1) daytime without sensory stimulations (no video-tape); 2) daytime with poor sensory stimulations (uninteresting video-tape film); 3) daytime with rich sensory stimulations (interesting chosen movie on video-tape); 4) night-time without sensory stimulations (no video-tape); 5) night-time with poor sensory stimulations (uninteresting video-tape). During the day, hedonic ratings decreased with time in the no- and uninteresting video-tape film conditions (p<0.01), but increased with the chosen movie (p<0.05). During the night, hedonic ratings decreased similarly to daytime ratings with the uninteresting video-tape film (p<0.01) but rose in the no-video-tape environment (p<0.01). The time course of motivation to leave the environment mirrored that of hedonic ratings. Changes in hedonic ratings as well as motivation to leave the environment correlated with the state of tiredness in the day-no-video and night-no-video situations (r=0.541 and r= 0.593; p<0.01). Thus, alliesthesia occurred in visual and auditory sensations that originated from the environment, and motivated behavior that was not consummatory. Such results suggest that alliesthesia is a general property of all sensations, and emphasizes the fundamental role of pleasure in motivation for all behaviors. PMID- 17399747 TI - Phenylethanoids, iridoids and a spirostanol saponin from Veronica turrilliana. AB - From the aerial parts of Veronica turrilliana two phenylethanoid glycosides, turrilliosides A and B and a steroidal saponin, turrillianoside were isolated and their structures elucidated as beta-(3,4-dihydroxyphenyl)ethyl-4-O-E-caffeoyl-O [beta-glucopyranosyl-(1-->4)-alpha-rhamnopyranosyl-(1-->6)]-beta-glucopyranoside, beta-(3,4-dihydroxyphenyl)ethyl-4-O-E-caffeoyl-[6-O-E-feruloyl-beta glucopyranosyl-(1-->4)-alpha-rhamnopyranosyl-(1-->6)]-beta-glucopyranoside and (23S,25S)-12beta,23-dihydroxyspirost-5-en-3beta-yl O-alpha-rhamnopyranosyl-(1- >4)-beta-glucopyranoside, respectively. Furthermore, eight known glucosides are reported namely, catalpol, catalposide, verproside, amphicoside, isovanilloylcatalpol, aucubin, arbutin, and 6-O-E-caffeoylarbutin, the latter two for the first time in the genus Veronica. The two phenylethanoid glycosides were found to be potent DPPH radical scavengers. All of the tested compounds were inactive against the representative species of fungi and bacteria. PMID- 17399748 TI - Anti-inflammatory and immunosuppressive compounds from Tripterygium wilfordii. AB - The extract of Tripterygium wilfordii Hook F. (TwHF), which showed anti inflammatory and immunosuppressive activities in human clinical trials for rheumatoid arthritis, was subjected to the activity-guided fractionation and spectroscopic characterization of bioactives. A tetrahydrofuran lignan, tripterygiol (1), and eight known compounds, all capable of suppressing pro inflammatory gene expression were identified. Most of the pharmacological activity of the extract can be attributed to triptolide, its most abundant and active component, with some contribution from tripdiolide. PMID- 17399749 TI - 20S proteasome and accumulation of oxidized and ubiquitinated proteins in maize leaves subjected to cadmium stress. AB - In order to examine the possible involvement of the 20S proteasome in degradation of oxidized proteins, the effects of different cadmium concentrations on its activities, protein abundance and oxidation level were studied using maize (Zea mays L.) leaf segments. The accumulation of carbonylated and ubiquitinated proteins was also investigated. Treatment with 50 microM CdCl(2) increased both trypsin- and PGPH-like activities of the 20S proteasome. The incremental changes in 20S proteasome activities were probably caused by an increased level of 20S proteasome oxidation, with this being responsible for degradation of the oxidized proteins. When leaf segments were treated with 100 microM CdCl(2), the chymotrysin- and trypsin-like activities of the 20S proteasome also decreased, with a concomitant increase in accumulation of carbonylated and ubiquitinated proteins. With both Cd(2+) concentrations, the abundance of the 20S proteasome protein remained similar to the control experiments. These results provide evidence for the involvement of this proteolytic system in cadmium-stressed plants. PMID- 17399750 TI - Iodus 40, salicylic acid, heptanoyl salicylic acid and trehalose exhibit different efficacies and defence targets during a wheat/powdery mildew interaction. AB - Prophylactic efficacies of Iodus 40 and salicylic acid (SA) against wheat powdery mildew caused by Blumeria graminis f. sp. tritici have been shown and compared with those of heptanoyl salicylic acid (HSA) and trehalose. Plantlets treated once exhibited 55%, 50%, 95%, and 38% protection levels, respectively. Two sprayings increased these levels up to 60%, 65%, 100%, and 60%, respectively. Biological effects of these resistance inducers on reactive oxygen species (ROS) metabolism and lipid peroxidation were also investigated. We found clear differences in the extent and the type of induced responses, with HSA exhibiting both the most numerous and the highest effects. HSA and SA induced a 5.5-fold increase of whole cell DAB staining due to hydrogen peroxide accumulation, whereas Iodus 40 and trehalose increased staining intensity at the penetration sites only. However, these effects were not correlated with any modification of catalase (CAT), oxalate oxidase (OXO) or lipoxygenase (LOX) activities, except for HSA which decreased CAT in non-inoculated conditions and increased LOX in infectious conditions. HSA also induced an increase in the rate of lipid peroxidation, whereas Iodus 40 induced a decrease. The effects of the inducers on germinating conidia and wheat epidermal cells responding to fungal penetration were also investigated. Papilla-linked autofluorescence was affected by SA and Iodus 40 whereas germination was slightly altered by Iodus 40. The newly described protective efficacies and the partial, distinct and non-overlapping activities of these inducers on the wheat/powdery mildew interaction are discussed. PMID- 17399751 TI - Functional genomics and the biosynthesis of artemisinin. AB - Artemisinin, a sesquiterpene lactone endoperoxide derived from the glandular secretory trichomes (GSTs) of Artemisia annua, provides the basis for the most effective treatments of malaria. The biology and biochemistry of GSTs of the Asteraceae and their biosynthesis of isoprenoids is reviewed. Recent efforts to understand the biosynthesis of artemisinin in A. annua GSTs are discussed in detail. This includes the development in the authors' laboratory of an expressed sequence tag (EST) approach to identifying the relevant biosynthetic genes using isolated GST as a source of mRNA. This has lead to the isolation of a cDNA encoding CYP71AV1, a multifunctional cytochrome P450 which catalyzes multiple oxidations of the sesquiterpene intermediate amorpha-4,11-diene to artemisinic acid. Further biochemical and molecular genetic work is required to elucidate the precise route from artemisinic alcohol to artemisinin and to engineer more efficient low cost production of artemisinin-based antimalarial drugs. PMID- 17399752 TI - Increases in morbid obesity in the USA: 2000-2005. AB - BACKGROUND: It is well known that citizens of developed countries are more likely to be overweight than they were 20 years ago. The most serious health problems are not associated with overweight or moderate obesity, however, but with clinically severe or morbid obesity (e.g. more than 100 pounds (45kg) overweight). There is no reason to expect that morbid obesity trends parallel overweight or moderate obesity. If morbid obesity is a rare pathological condition that has biological causes, the more than 10-fold increase in bariatric surgery procedures over the past eight years in the USA could have even lowered the prevalence of morbid obesity-and may very well stem the problem in other countries. OBJECTIVE: To estimate trends for extreme weight categories (BMI>40 and >50) for the period between 1986 and 2005 in the USA, and to investigate whether trends have changed since 2000. METHODS: Data from The Behavioral Risk Factor Surveillance System (a random-digit telephone survey of the household population of the USA), for the period from 1986 to 2005, were analysed. The main outcome measure was body mass index (BMI), calculated from self-reported weight and height. RESULTS: From 2000 to 2005, the prevalence of obesity (self-reported BMI over 30) increased by 24%. However, the prevalence of a (self-reported) BMI over 40 (about 100 pounds (45kg) overweight) increased by 50% and the prevalence of a BMI over 50 increased by 75%, two and three times faster, respectively. The heaviest BMI groups have been increasing at the fastest rates for 20 years. CONCLUSIONS: The prevalence of clinically severe obesity is increasing at a much faster rate among adults in the USA than is the prevalence of moderate obesity. This is consistent with the public health idea that the population weight distribution is shifting, which disproportionately increases extreme weight categories. Because comorbidities and resulting service use are much higher among severely obese individuals, the widely published trends for overweight/obesity underestimate the consequences for population health. The aggressive and costly expansion of bariatric surgery in recent years has had no visible effect on containing morbid obesity rates in the USA. PMID- 17399753 TI - [Cervicofacial cellulitis revealing cutaneous lymphomas]. AB - INTRODUCTION: The cervicofacial localization of cutaneous lymphomas is rare. These lymphomas usually present as a long-lasting and treatment-refractory papule or nodule. Lymphomas can also be revealed by cervicofacial cellulitis. CASES: We report 2 cases of cervicofacial cellulitis revealing a cutaneous lymphoma. The diagnosis was proved by multiple biopsies, performed because there was no clinical improvement in spite of an aggressive and adequate antibiotherapy. Our 2 patients were treated by radio and chemotherapy. DISCUSSION: Cutaneous lymphomas are lymphocytic proliferations stemming from cutaneous lymphoid tissue, without nodal, medullary, or visceral localization. Their clinical presentation is quite polymorphic, and cellulitis is one of the modes of revelation, especially forehead and neck localization. They have no portal of entry and are resistant to treatment. The diagnosis relies on histology, and biopsies must be performed if there is a suspicion of lymphoma. The treatment is radio and chemotherapy, and the evolution depends on the tumoral stage. PMID- 17399754 TI - Comment on: What do community-based dengue control programmes achieve? A systematic review of published evaluations. PMID- 17399755 TI - Quality of reclaimed waters: a public health need for source tracking of wastewater-derived protozoan enteropathogens in engineered wetlands. AB - Demand for high-quality drinking and recreational water rises exponentially owing to global demographic growth in the human population, reinforcing an urgent need for microbiologically safe reclaimed water. However, constructed wetlands, implemented into municipal wastewater treatment, may not provide substantial remediation for human protozoan enteropathogens such as Cryptosporidium, Giardia and human-virulent microsporidia. Improving reclaimed water quality by lowering faecal coliforms is not a sound solution for these pathogens. Current advances in molecular technology can benefit public health in developing and developed countries by changing the conceptual research framework for wastewater-receiving wetlands from 'pathogen removal' to 'pathogen source tracking' efforts. PMID- 17399756 TI - Development of malaria blood-stage vaccines: learning from mosquitoes. AB - If current methods of vaccine development for malaria continue to result in vaccines with only relatively limited degrees of protection, what is the alternative? Here, a totally different approach to blood-stage vaccine research is suggested, focusing on malarial immunity as it develops in macaque monkeys, but using methodology already well established in mosquito research. PMID- 17399757 TI - Sex differentiation pattern in the annual fish Austrolebias charrua (Cyprinodontiformes: Rivulidae). AB - Sex differentiation process, determination of sexual strategy, and gametogenesis of the annual fish Austrolebias charrua are established. Evidence of histological sex differentiation in an antero-posterior gradient was observed in pre-hatching stages. Sexual strategy corresponds to the "differentiated gonochoric" pattern. Histological analyses of adult gonads showed an asynchronous spawning mode for females and continuous spawning for males. Mature oocytes presented fluid yolk. Testis organization corresponded to a restricted spermatogonial model. Herein, we report the ultrastructural organization of the vitelline envelope and the main features of the sperm of A. charrua. Taking together these results also contribute to phylogenetic studies and provide base line data to propose A. charrua as a biomonitor of contamination in a protected area. PMID- 17399758 TI - Mercury species in lymphoid and non-lymphoid tissues after exposure to methyl mercury: correlation with autoimmune parameters during and after treatment in susceptible mice. AB - Methylmercury (MeHg) is present in the environment as a result of the global cycling of mercury, although anthropogenic sources may dramatically increase the availability in confined geographical areas. Accumulation of MeHg in the aquatic food chain is the dominating way of exposure in mammals, which accumulate MeHg in all organs, including the brain. Demethylation has been described in the organs, especially in phagocytic cells, but mainly in the flora of the intestinal tract. While most of the inorganic mercury (Hg(2+)) formed in the intestine is excreted, a fraction is reabsorbed which together with the local demethylation increases the organ Hg(2+) concentration. MeHg is a well-known immunosuppressive agent, while Hg(2+) is associated with immunostimulation and autoimmunity especially in genetically susceptible rodents, creating a syndrome, i.e. mercury-induced autoimmunity (HgIA). This study aimed at exploring the effect of MeHg with regard to HgIA, and especially the immunological events after stopping treatment, correlated with the presence of MeHg and Hg(2+) in the organs. Treatment of A.SW mice for 30 days with 4.2 mg MeHg/L drinking water (corresponding to approximately 420 microg Hg/kg body weight/day) caused all the HgIA features observed after primary treatment with inorganic Hg, except systemic immune complex deposits. The total Hg concentration was 5-fold higher in the kidneys as compared with lymph nodes, but the fraction of Hg(2+) was similar (17-20%). After stopping treatment, the renal and lymph node MeHg concentration declined according to first order kinetics during the initial 4-6 weeks, but then slower. A similar decline in the organ Hg(2+) concentration occurred during the initial 2 weeks after stopping treatment but then ceased, causing the Hg(2+) concentration to exceed that of MeHg in the lymph nodes and kidneys after 3 and 8 weeks, respectively. The selective increase in lymph node Hg(2+) fraction is likely to be due to demethylation of MeHg in the macrophage-rich lymphoid tissue. The major autoantibody in HgIA, anti-fibrillarin antibodies, tended to increase during the initial 6 weeks after stopping treatment, while all other HgIA features including antichromatin antibodies declined to control levels after 2-4 weeks. This indicates differences in either dose requirement or induction mechanisms for the different HgIA parameters. The selective accumulation of Hg(2+) in lymph nodes following MeHg treatment should be taken into account when the effect of MeHg on the immune system is evaluated. PMID- 17399759 TI - Sensitization of vascular smooth muscle cell to TNF-alpha-mediated death in the presence of palmitate. AB - Saturated free fatty acids (FFAs), including palmitate, can activate the intrinsic death pathway in cells. However, the relationship between FFAs and receptor-mediated death pathway is still unknown. In this study, we have investigated whether FFAs are able to trigger receptor-mediated death. In addition, to clarify the mechanisms responsible for the activation, we examined the biochemical changes in dying vascular smooth muscle cell (VSMC) and the effects of various molecules to the receptor-mediated VSMC death. Tumor necrosis factor (TNF)-alpha-mediated VSMC death occurred in the presence of sub-cytotoxic concentration of palmitate as determined by assessing viability and DNA degradation, while the cytokine did not influence VSMC viability in the presence of oleate. The VSMC death was inhibited by the gene transfer of a dominant negative Fas-associated death domain-containing protein and the baculovirus p35, but not by the bcl-xL or the c-Jun N-terminal kinase (JNK) binding domain of JNK interacting protein-1, in tests utilizing recombinant adenoviruses. The VSMC death was also inhibited by a neutralizing anti-TNF receptor 1 antibody, the caspase inhibitor z-VAD, and the cathepsin B inhibitor CA074, a finding indicative of the role of both caspases and cathepsin B in this process. Consistent with this finding, caspase-3 activation and an increase in cytosolic cathepsin B activity were detected in the dying VSMC. Palmitate inhibited an increase of TNF-alpha-mediated nuclear factor kappa B (NF-kappaB) activity, the survival pathway activated by the cytokine, by hindering the translocation of the NF-kappaB subunit of p65 from the cytosol into the nucleus. The gene transfer of inhibitor of NF-kappaB predisposed VSMC to palmitate-induced cell death. To the best of our knowledge, this study is the first report to demonstrate the activation of TNF-alpha-mediated cell death in the presence of palmitate. The current study proposes that FFAs would take part in deleterious vascular consequences of such patients with elevated levels of FFAs as diabetics and obese individuals via the triggering of receptor-mediated death pathways of VSMC. PMID- 17399761 TI - Fate of pharmaceutical and personal care products (PPCPs) during anaerobic digestion of sewage sludge. AB - The behaviour of 13 pharmaceutical and personal care products (PPCPs) has been studied during anaerobic digestion of sewage sludge: two musks (Galaxolide and Tonalide), one tranquilliser (Diazepam), one anti-epileptic (Carbamazepine), three anti-phlogistics (Ibuprofen, Naproxen and Diclofenac), two antibiotics (Sulfamethoxazole and Roxithromycin), one X-ray contrast medium (Iopromide) and three oestrogens (Estrone, 17beta-oestradiol and 17alpha-ethinyloestradiol). Two parallel processes have been carried out, one in mesophilic range (37 degrees C) and the other in thermophilic range (55 degrees C). The influence of temperature and sludge retention time (SRT) has been analysed. Among the substances considered, the higher removal efficiencies were achieved for the antibiotics, natural oestrogens, musks and Naproxen. For the other compounds, the values ranged between 20% and 60%, except for Carbamazepine, which showed no elimination. The removal of oestrogens, Diazepam and Diclofenac occurred after sludge adaptation. In general, no influence of SRT and temperature on PPCPs removal was observed. Considering the difficulty of obtaining reliable PPCPs concentrations, especially those corresponding to the fractions sorbed onto sludge, a methodology to validate the experimental data has been developed and successfully applied. PMID- 17399760 TI - Polyomavirus JC infects human brain microvascular endothelial cells independent of serotonin receptor 2A. AB - Although human polyomavirus JC (JCV) is known to cause progressive multifocal leukoencephalopathy (PML) in immunocompromised individuals, the mechanism by which JCV crosses the blood-brain barrier (BBB) remains unclear. To test our hypothesis that cell-free JCV gains entry into the brain by infecting endothelial cells, we inoculated human brain microvascular endothelial (HBMVE) cells with 50 HAU (1.33+/-0.27 x 10(7) genome copies) of JCV(Mad1) and analyzed the expression of early and late viral genes and proteins by immunocytochemistry, quantitative real-time PCR (qPCR), quantitative real-time reverse transcriptase PCR (qRT-PCR) and immunoprecipitation followed by Western blotting. JCV infected and replicated efficiently in HBMVE cells and produced infectious virions several hundred fold higher than the infecting inoculum. HBMVE cells in vitro did not express serotonin receptor 2A (5HT(2A)R), and 5HT(2A)R blockers did not prevent JCV infection of HBMVE cells. Collectively, our data indicate that the productive in vitro infection of HBMVE cells by JCV is independent of 5HT(2A)R. PMID- 17399762 TI - Mathematical model for predicting microbial reduction and transport of arsenic in groundwater systems. AB - A mathematical model was developed for describing the transport of arsenic, coupled with microbially-mediated biogeochemical processes. The biogeochemical characteristics of arsenic reactive transport processes were investigated in both batch and column tests, which showed that As(V) was reduced to As(III) by Shewanella sp., with the reduced arsenic species subsequently removed by precipitation. The breakthrough data obtained from the column experiments were used for the calibration of the arsenic reactive transport model. The reactive transport model, which only incorporated microbial reduction processes, showed a large discrepancy in predicting the observed As(III) concentration profiles, particularly later in the experiments. However, the model matched the experimental data much better with the inclusion of a term describing the precipitation process. Our results indicated that the precipitation reaction can be a major sink during microbially-mediated arsenic reactive transport. The proposed model provides a useful framework for predicting the transport of arsenic in saturated groundwater aquifers. PMID- 17399763 TI - Sorption and transport of acetaminophen, 17alpha-ethynyl estradiol, nalidixic acid with low organic content aquifer sand. AB - The sorption and transport of three pharmaceutical compounds (acetaminophen, an analgesic; nalidixic acid, an antibiotic; and 17alpha-ethynyl estradiol, a synthetic hormone) were examined by batch sorption experiments and solute displacement in columns of silica, alumina, and low organic carbon aquifer sand at neutral pH. Silica and alumina were used to represent negatively-charged and positively-charged fractions of subsurface media. Column transport experiments were also conducted at pH values of 4.3, 6.2, and 8.2 for the ionizable nalidixic acid. The computer program UFBTC was used to fit the breakthrough data under equilibrium and nonequilibrium conditions with linear/nonlinear sorption. Good agreement was observed between the retardation factors derived from column model studies and estimated from equilibrium batch sorption studies. The sorption and transport of nalidixic acid was observed to be highly pH dependent, especially when the pH was near the pK(a) of nalidixic acid (5.95). Thus, near a compound's pK(a) it is especially important that the batch studies be performed at the same pH as the column experiment. While for ionic pharmaceuticals, ion exchange to oppositely-charged surfaces, appears to be the dominant adsorption mechanism, for neutral pharmaceuticals (i.e., acetaminophen, 17alpha-ethynyl estradiol) the sorption correlated well with the K(ow) of the pharmaceuticals, suggesting hydrophobically motivated sorption as the dominant mechanism. PMID- 17399764 TI - Development of an image analysis procedure for identifying protozoa and metazoa typical of activated sludge system. AB - A procedure for the semi-automatic identification of the main protozoa and metazoa species present in the activated sludge of wastewater treatment plants was developed. This procedure was based on both image processing and multivariable statistical methodologies, leading to the use of the image analysis morphological descriptors by discriminant analysis and neural network techniques. The image analysis program written in Matlab has proved to be adequate in terms of protozoa and metazoa recognition, as well as for the operating conditions assessment. PMID- 17399765 TI - Biosurfactant technology for remediation of cadmium and lead contaminated soils. AB - This research focuses on column experiments conducted to evaluate the potential of environmentally compatible rhamnolipid biosurfactant produced by Pseudomonas aeruginosa strain BS2 to remove heavy metals (Cd and Pb) from artificially contaminated soil. Results have shown that di-rhamnolipid removes not only the leachable or available fraction of Cd and Pb but also the bound metals as compared to tap water which removed the mobile fraction only. Washing of contaminated soil with tap water revealed that approximately 2.7% of Cd and 9.8% of Pb in contaminated soil was in freely available or weakly bound forms whereas washing with rhamnolipid removed 92% of Cd and 88% of Pb after 36 h of leaching. This indicated that di-rhamnolipid selectively favours mobilization of metals in the order of Cd>Pb. Biosurfactant specificity observed towards specific metal will help in preferential elution of specific contaminant using di-rhamnolipid. It was further observed that pH of the leachates collected from heavy metal contaminated soil column treated with di-rhamnolipid solution was low (6.60-6.78) as compared to that of leachates from heavy metal contaminated soil column treated with tap water (pH 6.90-7.25), which showed high dissolution of metal species from the contaminated soil and effective leaching of metals with treatment with biosurfactant. The microbial population of the contaminated soil was increased after removal of metals by biosurfactant indicating the decrease of toxicity of metals to soil microflora. This study shows that biosurfactant technology can be an effective and nondestructive method for bioremediation of cadmium and lead contaminated soil. PMID- 17399766 TI - Trends in cervical cancer survival in Europe, 1983-1994: a population-based study. AB - OBJECTIVE: To evaluate trends in survival from cervical cancer in Europe and in European countries participating in the EUROCARE study as a function of age, morphology and stage at diagnosis. METHODS: Relative survival and relative excess risk of death within 5 years of diagnosis, as a function of age, morphology and stage, among 73,022 women aged 15-99 years diagnosed during 1983-1994 and followed up to 1999 in each of 18 European countries participating in the EUROCARE study, using data from 34 population-based cancer registries. RESULTS: Overall five-year relative survival was 62%, rising by 2% during the period 1983 1994. The highest survival occurred in Northern and Western Europe and the lowest in Central Europe. Survival falls with age at diagnosis, but mainly for localised disease. Survival is higher for adenocarcinoma in younger women, but higher for squamous cell carcinoma in older women. The proportions of younger women, localised cancer and adenocarcinoma all increased. The main improvements in survival were for women under 65, and for metastatic disease. CONCLUSIONS: Survival in Europe has improved slowly but steadily, but the trend is not geographically uniform. Central European countries and the UK saw little or no improvement, and survival in those countries remains the lowest among participating countries in Europe. Further reduction of cervical cancer mortality in Europe may be expected from expansion of screening, and improvement in the treatment of older women, and of metastatic disease. PMID- 17399767 TI - Insulin-like growth factor 2 and its receptors (IGF 1R and IGF 2R/mannose 6 phosphate) in endometrial adenocarcinoma. AB - OBJECTIVE: To investigate the consequences of IGF proteins dysfunction in development of endometrial adenocarcinomas. METHODS: The expression of IGF 2 and IGF 1R was correlated with the expression of IGF 2R and apoptosis rate in 59 human endometrial adenocarcinomas, 10 endometrial hyperplasias and 7 normal tissues. The presence of mutations in the IGF 2R gene was followed in 46 adenocarcinomas. We also examined the effect of IGF 1 receptor blockage on cancer cell proliferation. In groups of either IGF 2-positive or IGF 2-negative tumors (stages III and IV) the expression of IGF 1 and IGF 1R was correlated with cell proliferation index and telomerase activity. RESULTS: The expression of IGF 2 and IGF 1R was much higher in malignant tissue of stages III and IV than in tumors of stages I and II and normal or hyperplastic endometrium. This correlated with a decreased apoptosis rate and IGF 2R expression. Eight adenocarcinomas expressed biallelic mutation of the IGF 2R gene. The specific inhibition of IGF 1R and IGF 2 decreased tumor cell proliferation in IGF 2/IGF 1R-positive tumors. Furthermore, the positive correlation between increased expression of IGF 1 and IGF 1R proteins and increased telomerase activity and cell proliferation index was found in both IGF 2-negative and IGF 2-positive tumors. CONCLUSION: Our data suggest that IGF 1, IGF 2 and their receptors are involved in the progression of endometrial adenocarcinomas. As cancer cell proliferation can be abrogated by blocking mRNA or protein products of these genes, tumors with extensive involvement of the IGF 2 pathway would be candidates for the therapeutics strategies aimed at interference with this pathway. PMID- 17399768 TI - An analysis of the demographics and outcome of vulvar cancer in California. PMID- 17399769 TI - Fertility-sparing radical abdominal trachelectomy for clear cell adenocarcinoma of the upper vagina: a case report. AB - BACKGROUND: Clear cell adenocarcinoma (CCA) of the vagina is traditionally treated with radical surgical resection with tailored postoperative radiation when indicated. Due to a bimodal distribution, women of reproductive age are frequently affected and could benefit from radical trachelectomy to preserve fertility. CASE: A 22 year old female was diagnosed with clinical stage I vaginal clear cell adenocarcinoma in the left fornix abutting the cervix. The patient desired future fertility; therefore, she underwent radical abdominal trachelectomy and upper vaginectomy. Twenty-eight months after initial surgery, she has no evidence of recurrence with regular menstrual cycles. CONCLUSION: For patients with CCA of the upper vagina, where removal of the cervix is necessary, a radical trachelectomy with upper vaginectomy should be considered to conserve fertility. PMID- 17399771 TI - Airway inflammation in elite swimmers. PMID- 17399772 TI - Is there any role for allergen avoidance in the primary prevention of childhood asthma? AB - In this article we discuss 3 hypotheses to attempt to understand why preventive measures thus far studied with the aim of preventing (or delaying) the development of asthma have shown such disappointing results. The most likely explanation is that the development of a multifactorial disease, such as asthma, is extremely difficult, if not impossible, to prevent by eliminating only one risk factor. In a meta-analysis we investigated the effect of a multifaceted and monofaceted intervention in 10 prospective birth cohorts of a total of 3473 children on a diagnosis of asthma. Multifaceted intervention studies had an odds ratio (OR) of 0.73 (95% CI, 0.55-0.97), whereas the monointervention studies had an OR of 1.22 (95% CI, 0.83-1.78) in patients younger than 5 years and an OR of 0.52 (95% CI, 0.32-0.84) versus 0.93 (95% CI, 0.66-1.31) in patients older than 5 years. We therefore hypothesize that studies with a multifaceted approach will have a much greater chance of being successful than studies using a monofaceted approach, with the latter being unlikely to yield a clinically relevant reduction of asthma. PMID- 17399773 TI - Natural rubber latex allergy among health care workers. PMID- 17399774 TI - Implications of the asthma guidelines for the clinician. PMID- 17399776 TI - beta-adrenergic receptor genotype and response to salmeterol. PMID- 17399777 TI - Bronchoalveolar lavage invariant natural killer T cells are not increased in asthma. PMID- 17399779 TI - Eosinophilic disorders. AB - Eosinophilic inflammatory responses occur in association with multiple disorders. Although the initial cause and the affected organs vary among the different eosinophilic disorders, there are only 2 major pathways that mediate eosinophilia: (1) cytokine-mediated increased differentiation and survival of eosinophils (extrinsic eosinophilic disorders), and (2) mutation-mediated clonal expansion of eosinophils (intrinsic eosinophilic disorders). Independent from the original trigger, the most common cause of eosinophilia is the increased generation of IL-5-producing T cells. In some cases, tumor cells are the source of eosinophil hematopoietins. The intrinsic eosinophilic disorders are characterized by mutations in pluripotent or multipotent hematopoietic stem cells leading to chronic myeloid leukemias with eosinophils as part of the clone. Here, we propose a new classification of eosinophilic disorders on the basis of these obvious pathogenic differences between the 2 groups of patients. We then discuss many known eosinophilic disorders, which can be further subdivided by differences in T-cell activation mechanisms, origin of the cytokine-producing tumor cell, or potency of the mutated stem cell. Interestingly, many subgroups of patients originally thought to have the idiopathic hypereosinophilic syndrome can be integrated in this classification. PMID- 17399780 TI - Salmeterol response and beta(2)-adrenergic receptor genotype. PMID- 17399781 TI - Longitudinal study on cat allergen exposure and the development of allergy in young children. AB - BACKGROUND: The influence of cat allergen exposure in early childhood on the development of sensitization and allergic diseases is complex. Little is known about the natural course of the sensitization development within individuals. OBJECTIVE: We investigated the association between cat allergen exposure in infancy and cat ownership and cat contact during childhood and the development of cat sensitization and allergic diseases up to age 6 years using a longitudinal analysis approach. METHODS: Overall, 2166 children from an ongoing birth cohort study were included in the analysis. House dust samples were collected 3 months after birth. Cat allergen levels were extracted. Blood samples were collected when the children were 2 and 6 years old. Information on the allergic symptoms of children and doctor-diagnosed allergic disease were collected at each follow-up using questionnaires. RESULTS: Cat allergen exposure in infancy was positively associated with sensitization at age 2 years but not at age 6 years. No associations existed between cat allergen exposure in infancy and allergic symptoms and diseases up to age 6 years. Cumulative allergen exposure from cat ownership and regular cat contact increased the risk of cat sensitization up to age 6 years. CONCLUSION: Cat allergen exposure in infancy increases the risk of sensitization development in early childhood but not in school-age children. Cumulative allergen exposure from cat ownership and regular cat contact during childhood contribute to sensitization development up to school age. CLINICAL IMPLICATIONS: Cat allergen avoidance at home alone might be not effective to prevent the development of allergic sensitization in young children. PMID- 17399782 TI - Respiratory viruses and acute asthma in children. PMID- 17399783 TI - Synthesis and mechanical properties of quaternary salts of chitosan-based films for food application. AB - A novel method is described to synthesize quaternary salts of chitosan with dimethylsulfate and subsequently cast films. In an attempt to improve both mechanical and hydrophobic characteristics, the chitosan was previously modified by N-alkylation, introducing 4, 8 and 12 carbons moieties into the polymeric chain. Analysis by FTIR and solid-state CP-MAS (13)C NMR spectroscopy confirmed the success of both alkylation and quaternization processes. The average degree of quaternization of these N-methylated derivatives was calculated to be 35%. DMA measurements indicated that chitosan and its derivative films are typically brittle materials, exhibiting similar non-linear viscoelastic behaviors. The films of unmodified chitosan have a very small strain (approximately 2.8%), though they were the most resistant films (Young's modulus=2283 MPa; tensile strength >44.0 MPa). In general, the alkyl-chitosan derivatives appear to be more plastic than chitosan films but less resistant, e.g., for butyl chitosan: maximum strain=13.1%; tensile strength=13.4 MPa and Young's modulus=171 MPa. Conversely the quaternization reaction increased the hardness of the parent sample, viz. for quaternary salt of dodecyl chitosan: maximum strain=2.6%; tensile strength=38.3 MPa and Young's modulus=1792 MPa. PMID- 17399784 TI - Ectopic neocartilage formation from predifferentiated human adipose derived stem cells induced by adenoviral-mediated transfer of hTGF beta2. AB - Chondrogenic potential of human adipose derived stem cells (hASCs) makes them a possible source of seeding cells for cartilage tissue engineering. In this study, chondrogenic differentiation of hASCs induced by transduction with replication deficient adenovirus carrying human transforming growth factor beta2 (Ad5-hTGF beta2) was demonstrated by RT-PCR, immunohistochemistry staining, biochemical and western blot analysis. To evaluate if the in vitro differentiated hASCs could keep their chondrocytic phenotype and produce neo-cartilage in vivo, predifferentiated hASCs were seeded in different scaffolds and implanted in subcutaneous pockets on the dorsum of nude mice. After 4 and 12 weeks culture in vivo, specimens were harvested and examined by histological and immunohistochemical analysis, cartilage-like tissue formation was only found in alginate gel and PLGA/alginate compound groups, in PLGA group, fibrous tissues and angiogenesis ingrowth were observed. These findings demonstrated that adenovirus-mediated hTGF beta2 gene transfer could induce hASCs into a chondrogenic lineage in vitro, however, this predifferentiation did not guarantee ectopic cartilage formation in vivo unless appropriate three-dimensional scaffolds were used as the cell carry vehicles. PMID- 17399785 TI - Addressing symptoms of depression in foster caregivers may improve the quality of care children receive. PMID- 17399786 TI - Resilience to adult psychopathology following childhood maltreatment: evidence from a community sample. AB - OBJECTIVE: Child abuse is an important risk for adult psychiatric morbidity. However, not all maltreated children experience mental health problems as adults. The aims of the present study were to address the extent of resilience to adult psychopathology in a representative community sample, and to explore predictors of a good prognosis. METHODS: Data are drawn from a follow-up of the Isle of Wight study, an epidemiological sample assessed in adolescence and at midlife. Ratings of psychiatric disorder, peer relationships and family functioning were made in adolescence; adult assessments included a lifetime psychiatric history, personality and social functioning assessments, and retrospective reports of childhood sexual and physical abuse. RESULTS: Ten percent of individuals reported repeated or severe physical or sexual abuse in childhood. Prospective measures revealed increased rates of adolescent psychiatric disorders in this group. Rates of adult psychopathology were also high. A substantial minority of abused individuals reported no mental health problems in adult life. Resilience of this kind was related to perceived parental care, adolescent peer relationships, the quality of adult love relationships, and personality style. CONCLUSION: Good quality relationships across childhood, adolescence and adulthood appear especially important for adult psychological well being in the context of childhood abuse. PMID- 17399788 TI - Health implications of radon distribution in living rooms and bedrooms in U.K. dwellings--a case study in Northamptonshire. AB - Environmental radon exposure of residents of domestic premises in the United Kingdom (UK) and elsewhere in Europe is estimated on the basis of the measured radon concentrations in, and the relative occupancies of, the principal living room and bedroom. While studies on radon concentration variability in the individual units in apartment blocks in various countries have been described, little data has been reported on variability in two-storey single-family dwellings, and the majority of extant studies consolidate living room and bedroom data early in the analysis. To investigate this further, detailed analysis was made of radon concentration data from a set of thirty-four homes situated in areas of Northamptonshire known to exhibit high radon levels. All homes were of typical UK construction of brick/block/stone walls under a pitched tile/slate roof. Approximately 50% of the sample were detached houses, the remainder being semi-detached (duplex) or terraced (row-house). Around 25% of the sample possessed cellars, while 12% were single-storey dwellings (bungalows), reflecting the typical incidence of this type of dwelling in England. In the two-storey homes, all monitored bedrooms were on the upper floor. Distribution of the ratios of bedroom/living room radon concentrations (BR/LR ratio) in individual properties was left-skewed (mean 0.67, median 0.73, range 0.05-1.05) with a tail extending to just above 1.0. The mean is consistent with the outcome of earlier extensive studies in England, while the variability depends principally on the characteristics of the property, and not on seasonal factors. In a small set of homes, the BR/LR ratio was anomalously low, (mean 0.3). BR/LR ratios in single storey homes clustered around a value of 1.0, indicating that house design, rather than lifestyle, is the dominant factor in determining bedroom radon concentrations. Homes with higher mean annual radon concentrations showed lower BR/LR ratios, supporting our proposal that, in some homes, radon emanation from building materials may comprise a significant component of the overall radon level. PMID- 17399789 TI - Comparative overview of indoor air quality in Antwerp, Belgium. AB - This comprehensive study, a first in Belgium, aimed at characterizing the residential and school indoor air quality of subgroups that took part in the European Community Respiratory Health Survey and the International Study of Asthma and Allergy in Childhood [Masoli M, Fabian D, Holt S, Beasley R. Global Burden of Asthma, Medical Research Institute of New Zealand, University of Southampton; 2004.] questionnaire-based asthma and related illnesses studies. The principal aim was to perform a base-line study to assess the indoor air quality in Antwerp in terms of various gaseous and particulate pollutants. Secondly, it aimed to establish correlations between these pollutants investigated, the pollutant levels in the indoor and outdoor micro-environments, findings of the previous questionnaire-based studies and an epidemiological study which ran in conjunction with this study. Lastly, these results were compared and evaluated with current indoor and ambient guidelines in various countries This paper presents selected results on PM1, PM2.5 and PM10 mass concentrations and elemental C estimates as black smoke, as well as gaseous NO(2), SO(2), O(3) and BTEX concentrations of 18 residences and 27 schools. These are related to current guidelines of Flanders, Germany, Norway, China and Canada and evaluated with reference to selected similar studies. It was found that indoor sources such as tobacco smoking and carpets, the latter causing re-suspension of dust, are responsible for elevated indoor respirable particulate matter and place school children and residents at risk. Both PM2.5 and PM10 equalled or exceeded the current guidelines adopted by Flanders, noting that 12-h and 24-h PM2.5 were compared with an annual limit value. Indoor and ambient NO(2) concentrations in the school campaign were higher than the annual EU ambient norm. The other studied pollutant levels were below the current guidelines. PMID- 17399790 TI - Defective complement control of factor H (Y402H) and FHL-1 in age-related macular degeneration. AB - The common variant in the human complement Factor H gene (CFH), with Tyr402His, is linked to age-related macular degeneration (AMD), a prevalent disorder leading to visual impairment and irreversible blindness in elderly patients. Here we show that the risk variant CFH 402His displays reduced binding to C reactive protein (CRP), heparin and retinal pigment epithelial cells. This reduced binding can cause inefficient complement regulation at the cell surface, particularly when CRP is recruited to injured sites and tissue. In addition, we identify the Factor H-like protein 1 (FHL-1), an alternative splice product of the CFH gene as an additional protein that includes the risk residue 402, and thus confers risk for AMD. FHL-1 is expressed in the eye and the FHL-1 402His risk variant shows similar reduced cell binding and likely reduced complement regulatory functions on the cell surface. CFH and FHL-1 may act in concert in the eye and the reduced surface binding may result in inappropriate local complement control, which in turn can lead to inflammation, disturbance of local physiological homeostasis and progression to cell damage. As a consequence, these processes may lead to AMD pathogenesis. PMID- 17399791 TI - Coagulation, coma, and outcome in bacterial meningitis--an observational study of 38 adult cases. AB - OBJECTIVES: The purpose of this study was to evaluate the epidemiology of intravascular coagulation in bacterial meningitis and to recognise the associations with disease severity and outcome. METHODS: Thirty-eight consecutively admitted adult patients with microbiologically proven bacterial meningitis were observed prospectively for platelets count (PLT), platelets decline (dPLT), prothrombin ratio (PTr), INR, and D-dimer levels during the first three days in relation to disease severity (Glasgow Coma Scale--GCS, APACHE-III) and outcome (Glasgow Outcome Scale--GOS). RESULTS: The prevalence of activated coagulation measured by abnormal laboratory results varied respectively: PTr- 30%, INR--36%, PLT--38%, dPLT--50%, and D-dimer--88%. Patients with GCS <9 at admission presented with laboratory results suggesting triggered coagulation: dPLT 48 vs. 15%/day (p=0.0246), INR 1.6 vs. 1.12 (p=0.0014), PTr 76 vs. 93% (p=0.0020). An unfavourable outcome (GOS 1-4) was observed in 42% of patients and was associated with: PLT <170 or >265 G/L (OR--24.4; p=0.0006), PTr <82% (OR- 5.00; p=0.0388), INR >1.1 (OR--5.04; 0.0336), and D-dimer >850 ng/ml (OR--24.0; p=0.0033). CONCLUSIONS: Coagulation was activated in a majority of patients with bacterial meningitis and related to coma and unfavourable outcome. PMID- 17399792 TI - Explaining sex differences in infants' preferences for groups. AB - Social organization of a species influences myriad facets of individuals' behavior. Much research indicates that human social organization consists of males in large groups and females in smaller groups or interacting with individuals. This study analyzed the initial factors that produce greater preferences for groups by human male versus female infants. To this end, using a looking preference paradigm, fifty-nine 6-8-month-old infants viewed individual versus group images of actual children. On the basis of several controls, results demonstrated that male more than female infants are attracted to the complex level of stimulation provided by groups. Discussion centers on further identifying male versus female patterns of group interaction from a perceptual and cognitive standpoint. PMID- 17399793 TI - Neurotoxins from invertebrates as anticonvulsants: from basic research to therapeutic application. AB - Invertebrate venoms have attracted considerable interest as a potential source of bioactive substances, especially neurotoxins. These molecules have proved to be extremely useful tools for the understanding of synaptic transmission events, and they have contributed to the design of novel drugs for the treatment of neurological disorders and pain. In this context, as epilepsy involves neuronal substrates, which are sites of action of many neurotoxins; venoms may be particularly useful for antiepileptic drug (AED) research. Epilepsy is a chronic disease whose treatment consists of controlling seizures with antiepileptics that very often induce strong undesirable side effects that may limit treatment. Here, we review the vast, but yet unexplored, world of neurotoxins from invertebrates used as probes in pharmacological screening for novel and less toxic antiepileptics. We briefly review (1) the molecular basis of epilepsy, as well as the sites of action of commonly used anticonvulsants (we bring a comprehensive review of the elements from invertebrate venoms which are mostly studied in neuroscience research and may be useful for drug development); (2) peptides from conus snails; (3) peptides and polyamine toxins from spiders and wasps; and (4) peptides from scorpions. PMID- 17399794 TI - Inhibition of mitogen-activated protein kinase and stimulation of Akt kinase signaling pathways: Two approaches with therapeutic potential in the treatment of neurodegenerative disease. AB - The neurodegenerative diseases of adulthood, including Alzheimer's disease (AD) and Parkinson's disease (PD), pose an enormous and growing public health burden. Although effective symptomatic treatments exist for PD, and, to a lesser extent, for AD, there is no therapy for these disorders which will forestall their progression. With the rise of the concept of programmed cell death (PCD) came the realization that even in the absence of complete knowledge of proximate causes neuroprotection may nevertheless be possible by targeting the pathways of PCD. One set of signaling pathways that have been implicated in cell death are the mitogen-activated protein kinase (MAPK) pathways. The possibility of blocking these pathways and thereby providing neuroprotection has recently been put to the test in a clinical trial of a mixed lineage kinase inhibitor in the treatment of PD. Unfortunately, this trial failed to demonstrate a protective effect. Based on considerations related to the implementation of the trial, it would be premature to conclude that inhibition of MAPK signaling is a failed strategy. In spite of these negative results, the MAPK and related kinase pathways retain their importance as potential targets in PD. In relation to pathogenesis, the discovery of mutations in the mixed lineage kinase (MLK)-like kinase leucine-rich repeat kinase 2 (LRRK2) suggests a role for these kinases in regulating the viability of dopamine neurons. In relation to treatment, the survival signaling kinase Akt has been demonstrated in vivo to mediate striking neurotrophic and antiapoptotic effects. Thus, it is likely that therapeutic targets related to these kinase signaling pathways will emerge. PMID- 17399795 TI - A colorimetric assay method to measure acetyl-CoA synthetase activity: application to woodchuck model of hepatitis virus-induced hepatocellular carcinoma. AB - A new spectrophotometric method for quantitation of acetyl-CoA synthetase (ACAS) activity is developed. It has been applied for ACAS assay in the liver tissues of a woodchuck model of hepatitis virus-induced hepatocellular carcinoma (HCC). The assay is based on the established pyrophosphate (PPi) detection system. ACAS activity is indexed by the amount of PPi, the product of ACAS reaction system of activated form of acetate (acetyl-CoA) with ACAS catalysis. PPi is determined quantitatively as the amount of chromophore formed with molybdate reagent, 1 amino-2-naphthol-4-sulfonic acid in bisulfite and 2-mercaptoethanol. PPi reacts with molybdate reagent to produce phosphomolybdate and PPi-molybdate complexes. 2 mercaptoethanol is responsible for color formation which has the peak absorbance at 580 nm. This method was sensitive from 1 to 20 nmol of PPi in a 380-mul sample (1-cm cuvette). A ten-fold excess of Pi did not interfere with the determination of PPi. To study the major metabolic pathways of imaging tracer [1-(11)C]-acetate in tumors for detection of HCC by Positron Emission Tomography (PET), the activity of one of the key enzymes involved in acetate or [1-(11)C]-acetate metabolism, ACAS was assayed by this newly developed assay in the tissue samples of woodchuck HCCs. A significant increase of ACAS activity was observed in the liver tissues of woodchuck HCCs as compared with neighboring regions surrounding the tumors (P<0.05). The respective ACAS activities in the subcellular locations were also significantly higher in HCCs than in the surrounding tissues (P<0.05) (total soluble fraction: 876.61+/-34.64 vs. 361.62+/-49.97 mU/g tissue; cytoplasmic fraction: 1122.02+/-112.39 vs. 732.32+/-84.44 mU/g tissue; organelle content: 815.79+/-100.77 vs. 547.91+/-97.05 mU/ g tissue; sedimentable fragment: 251.92+/-51.56 vs. 90.94+/-18.98 mU/ g tissue). The finding suggests an increase in ACAS activity in the liver cancer of woodchuck models of HCC as compared to that in the normal woodchuck liver. The developed assay is rapid, simple and accurate and is suitable for the investigation of ACAS activity under physiologic and pathophysiologic conditions. PMID- 17399796 TI - Sample preparation for two-dimensional blue native/SDS polyacrylamide gel electrophoresis in the identification of Streptomyces coelicolor cytoplasmic protein complexes. AB - Ammonium sulfate precipitation was tested as a sample preparation step for BN PAGE analyses of S. coelicolor cytoplasmic protein complexes. A procedure of sample preparation compatible with two-dimensional BN/SDS-PAGE was established and used to visualize protein complexes. To validate the sample preparation procedure, representative protein complexes were identified. Several previously characterized protein complexes were rediscovered and their reported oligomeric states reconfirmed. In addition, we identified new but plausible interactions that have never been reported before. Our work provides useful reference for the wide application of BN-PAGE in protein interaction study. PMID- 17399797 TI - An auditory brain-computer interface (BCI). AB - Brain-computer interfaces (BCIs) translate brain activity into signals controlling external devices. BCIs based on visual stimuli can maintain communication in severely paralyzed patients, but only if intact vision is available. Debilitating neurological disorders however, may lead to loss of intact vision. The current study explores the feasibility of an auditory BCI. Sixteen healthy volunteers participated in three training sessions consisting of 30 2-3 min runs in which they learned to increase or decrease the amplitude of sensorimotor rhythms (SMR) of the EEG. Half of the participants were presented with visual and half with auditory feedback. Mood and motivation were assessed prior to each session. Although BCI performance in the visual feedback group was superior to the auditory feedback group there was no difference in performance at the end of the third session. Participants in the auditory feedback group learned slower, but four out of eight reached an accuracy of over 70% correct in the last session comparable to the visual feedback group. Decreasing performance of some participants in the visual feedback group is related to mood and motivation. We conclude that with sufficient training time an auditory BCI may be as efficient as a visual BCI. Mood and motivation play a role in learning to use a BCI. PMID- 17399798 TI - Off-line removal of TMS-induced artifacts on human electroencephalography by Kalman filter. AB - In this paper we present an off-line Kalman filter approach to remove transcranial magnetic stimulation (TMS)-induced artifacts from electroencephalographic (EEG) recordings. Two dynamic models describing EEG and TMS signals generation are identified from data and the Kalman filter is applied to the linear system arising from their combination. The keystone of the approach is the use of time-varying covariance matrices suitably tuned on the physical parameters of the problem that allow to model the nonstationary components of the EEG-TMS signal. This guarantees an efficient deletion of TMS-induced artifacts while preserving the integrity of EEG signals around TMS impulses. Experimental results show that the Kalman filter is more effective than stationary filters (Wiener filter) for the problem under investigation. PMID- 17399799 TI - Lower rates of depression in westernised Chinese in the US. AB - BACKGROUND: Low reported rates of depression in Chinese populations could reflect real or artefactual factors, and might be clarified by studying acculturated Chinese in western regions. We therefore sought to determine whether reported rates of depressive disorders differ in resident Chinese and matched non-Chinese controls in a large US community survey. METHOD: We accessed data from the US National Epidemiological Survey of Alcoholism and Related Conditions, involving 306 Chinese subjects and 306 matched non-Chinese subjects. RESULTS: The Chinese reported significantly lower lifetime and 12-month major depression rates, and a lower lifetime rate of dysthymia. Similar rates were quantified for Chinese born in the US and Chinese born overseas. The Chinese did not differ from controls in terms of recurrence rates of major depression. LIMITATION: Due to our matching analytic strategy, the reported statistics (e.g. prevalences) apply to our matched samples and should not be taken as estimates for the population. CONCLUSIONS: Findings indicate that westernisation does not eliminate differences in depression rates long described in Chinese regions, and favour a model whereby the Chinese have a lower vulnerability to depression onset. PMID- 17399800 TI - Soluble HLA-G molecules in follicular fluid: a tool for oocyte selection in IVF? AB - Currently, different approaches are used to select oocytes for in vitro fertilization (IVF) procedures, but they do not assure a significant association with the pregnancy outcome. Since several studies have proposed the expression of HLA-G antigens in early embryos to be a possible marker of elevated implantation rate, we have investigated the presence of soluble HLA-G molecules in 50 follicular fluids (FFs). The results have shown soluble HLA-G antigens (sHLA-G) in 19/50 (38%) FFs. Furthermore, we have related the presence of sHLA-G molecules in FFs to detection of the soluble antigens in culture supernatants of the corresponding fertilized oocyte, evidencing a significant relationship (p=1.3 x 10(-6); Fisher exact p-test). Specific ELISA and Western blot approaches identified both HLA-G5 and soluble HLA-G1 molecules in FFs while immunocytochemical analysis indicated polymorphonuclear-like and granulosa cells as responsible for production of sHLA-G1 and HLA-G5 molecules. In contrast, only sHLA-G1 antigens were detected in culture supernatants of fertilized oocytes. Overall, these results suggest a role for sHLA-G molecules in the ovulatory process and propose the FFs analysis for sHLA-G molecule presence as a useful tool for oocyte selection in IVF. PMID- 17399801 TI - Is filtering difficulty the basis of attentional deficits in schizophrenia? AB - The distractibility that schizophrenia patients display may be the result of a deficiency in filtering out irrelevant information. The aim of the current study was to assess whether patients with schizophrenia exhibit greater difficulty when task-irrelevant features change compared to healthy participants. Thirteen medicated outpatients with a diagnosis of schizophrenia and thirteen age- and parental education-matched controls performed a target selection task in which the task-relevant letter or the task-irrelevant features of color, and/or location repeated or switched. Participants were required to respond by pressing the appropriate key associated with the target letter. These patients with schizophrenia were slower when the task-relevant target letter switched than when it repeated. In contrast, schizophrenia patients performed similarly to controls when task-irrelevant information changed. Thus, we found no evidence that patients with schizophrenia were impaired in inhibiting irrelevant perceptual features. In contrast, changes in task-relevant features were problematic for patients relative to control participants. These results suggest that medicated outpatients who are mild to moderately symptomatic do not exhibit global impairments of feature processing. Instead, impairments are restricted to situations when task-relevant features vary. The current findings also suggest that when a course of action is not implied by an irrelevant feature, outpatients' behavior is not modulated by extraneous visual information any more than in healthy controls. PMID- 17399802 TI - Suction diathermy for adenoidectomy: complications and risk of recurrence. AB - OBJECTIVES: When introduced, suction coagulation was initially utilised for haemorrhage control following curettage of the adenoid pad. More recently the whole procedure has been performed using the technique. This study aims to report post-operative haemorrhage rates and risk of recurrence in adenoidectomy performed solely by suction diathermy in children. METHODS: A retrospective study of 1411 consecutive paediatric patients. Surgery was performed using suction diathermy. No patients were excluded. All patients were followed up. RESULTS: There were no cases of post-operative haemorrhage. 1.7% of patients remained symptomatic and underwent revision adenoidectomy. None required a third procedure. CONCLUSIONS: Re-growth of adenoid tissue may occur despite visualisation of the nasopharynx at the time of surgery. The incidence of re growth is similar to that reported in patients undergoing conventional adenoidectomy by curettage. Post-operative haemorrhage was not encountered in children having adenoidectomy by suction diathermy. The authors suggest suction diathermy as the most appropriate method for adenoidectomy in children. PMID- 17399803 TI - DNA extraction: an understudied and important aspect of HPV genotyping using PCR based methods. AB - Testing for the group of approximately 15 carcinogenic human papillomavirus (HPV) genotypes is an important adjunct to cytology. Because carcinogenic strengths of HPV types differ greatly, assays that permit identification of individual HPV genotypes are being introduced. Most HPV genotyping systems proposed for clinical use are PCR-based, depending heavily for validity on careful attention to numerous details. One understudied detail is the effect of different sample preparation methods including DNA extraction. This study examines the influence of DNA extraction on performance of a new PCR-based genotyping kit, the Roche LINEAR ARRAY HPV assay. When volume of sample extracted, DNA extraction methods and/or amount of DNA tested were varied, the HPV type results were reproducible for strong viral bands but not weak ones. Moreover, although the experiments were not comprehensive, they showed that the manufacturer-approved DNA extraction method might not be the best method for use in this assay. Because different "front end" protocols introduce variability into genotyping results, the authors urge laboratories not to vary methods for this assay without due consideration. The results suggest that companies carefully optimize DNA extraction methods prior to commercial introduction of their PCR-based genotyping assays destined for widespread clinical use. PMID- 17399804 TI - Individual variation in sleep and motor activity in rats. AB - We examined individual differences in sleep and motor activity across 2 consecutive days in rats. EEG and motor activity were recorded via telemetry in Wistar rats (n=29) for 48h under well-habituated conditions. Rats were grouped based on sleep amounts and stability across days (short [SS, n=7], intermediate [IS, n=15] and long [LS, n=7] sleep) and comparisons were conducted to determine group differences for measures of sleep and motor activity. We found that correlations across recording days were significant for all selected sleep measures and motor activity counts. Rankings for 24h total sleep time and non rapid eye movement sleep (NREM) were SS36 months from closure. RESULTS: Compared to 6 months after closure, a further improvement of peak oxygen uptake (p<0.001) and of the slope of ventilation/carbon dioxide production (p<0.001) was observed 3 years after the procedure, so that peak oxygen uptake appeared to be within the normal range in 23/29 patients (79%). Right ventricular short-axis (p<0.05) and long-axis (p<0.05) diameters further decreased beyond the 6-month period. The long-term improvement in exercise capacity correlated with pulmonary-to-systemic flow ratio (R=0.55, p=0.003) and with percentage decrease in RV short-axis diameter (R=0.59, p=0.002), but it did not correlate with age at closure (R=0.25, p=0.46). All patients who did not achieve a normal exercise capacity after 3 years from closure had a severely depressed pre-closure peak oxygen uptake (<50% of predicted). CONCLUSIONS: Adults who undergo transcatheter ASD closure may experience a further improvement in exercise capacity in the long term. The long term improvement in exercise capacity is associated to an improvement in cardiac form and function and is not influenced by age at closure. Even if the majority of patients may reach a normal exercise capacity after ASD closure, an abnormal exercise capacity may persist in those patients that had a peak oxygen uptake below 50% of predicted value before the procedure. PMID- 17399818 TI - Mild to moderate renal impairment is associated with increased left ventricular mass. AB - BACKGROUND: Mild to moderate renal impairment (MMRI) is associated with an excess cardiovascular mortality but the reason for this is unclear. Increased left ventricular mass (LVM) is associated with severe renal dysfunction and a higher cardiovascular mortality. However, whether increased LVM also occurs in patients with milder renal dysfunction is unknown. METHODS: 336 consecutive patients under investigation for chest pain who underwent concurrent coronary angiography, serum creatinine (Cr) and LVM assessment, were recruited from a university hospital in Shanghai, China. The main outcome measures were: LVM normalised for body surface area (NLVM), renal function determined by Cr, creatinine clearance (CrCl) (Cockroft-Gault equation) and glomerular filtration rate (GFR) (Modification of Diet in Renal Disease equation). RESULTS: Mean+/-SD age was 56.8+/-9.5 years, 282 (84%) were male, mean NLVM 107.1+/-36.8 g/m(2), Cr 0.94+/-0.22 mg/dL, CrCl 82.7+/ 21.7 mL/min/1.73 m(2) and GFR 89.4+/-24.6 mL/min/1.73 m(2). NLVM was related to Cr (r=-0.30), CrCl (r=0.19) and GFR (r=0.24) (all p<0.001). A multiple logistic regression model using quartile analysis of renal function showed that those patients with a Cr 1.06-2.00 mg/dL, CrCl 34.8-67.5 mL/min/1.73 m(2) and a GFR 36.2-73.4 mL/min/1.73 m(2) were significantly associated with increased NLVM, independent of other variables. Using definitions derived from the National Kidney Foundation Guidelines multiple regression analysis showed MMRI to be independently associated with increased NLVM: Cr 1.2-1.9 mg/dL, odds ratio (OR) 2.77 (CI 1.04-7.40) (p=0.04); CrCl 30-89.9 mL/min/1.73 m(2), OR 1.63 (CI 0.91 2.93) (p=0.11); GFR 30-89.9 mL/min/1.73 m(2), OR 1.76 (CI 1.07-2.90) (p=0.03). CONCLUSION: In patients being investigated for chest pain, MMRI is significantly and independently associated with increased LVM. PMID- 17399819 TI - Impact of socioeconomic status on outcome of a Brazilian heart transplant recipients cohort. AB - We studied the clinical course of 44 cardiac transplant recipients at our institution according to socioeconomic status (socioeconomic class level, educational level, household family income, dwelling, presence of caregiver, and national health insurance) before the procedure. Patients in the low socioeconomic status had a prognosis similar to that seen in patients in the median socioeconomic status. Thus, low socioeconomic status has no unfavorable impact on outcome of cardiac transplant recipients in Brazil. PMID- 17399820 TI - Intravascular ultrasound assessment of atherosclerotic plaque in the aorta. AB - BACKGROUND: Intravascular ultrasound (IVUS) has been used for assessment of the coronary arteries. However, few IVUS studies have been reported on the aorta. MATERIALS AND METHODS: To assess the presence of atherosclerotic disease in the aorta by the use of IVUS, 29 patients with heart disease (24 men and 5 women; mean age, 61+/-11 years) including ischemic heart disease (n=21) and valvular disease or dilated cardiomyopathy (n=8) were enrolled in the study. An IVUS catheter was inserted through the femoral artery, and IVUS images were obtained in the descending aorta (DA) at the level of the pulmonary artery bifurcation, and in the proximal and distal regions of the abdominal aorta (AA) at the level of the renal artery bifurcation. Percent plaque area (%PA) was calculated as vessel cross-sectional area surrounded by media minus lumen cross-sectional area divided by vessel cross-sectional area. RESULTS: %PAs differed significantly between the three aortic levels: DA, 14.9+/-5.5%; proximal AA, 19.0+/-6.9%; and distal AA, 28.3+/-9.7% (p<0.05). However, PA did not differ significantly between the three levels (DA, 94.7+/-38.0 mm2; proximal AA, 90.9+/-35.0 mm2; distal AA, 79.7+/-32.3 mm2). %PA and PA in the DA and proximal AA correlated with age (r=0.39-0.46, p<0.05), but not with coronary angiography findings or multiple risk factors. CONCLUSIONS: The aortic plaque is clearly observed by IVUS. The plaque is diffuse at the 3 levels, had little relationship with risk factor of arteriosclerosis or coronary artery disease, and aging affected the increase of plaque. PMID- 17399821 TI - Association of chronic subclinical inflammation with severity and progression of rheumatic valve disease. PMID- 17399822 TI - Six-minute walk test in Chagas cardiomyopathy. AB - We studied systematically, for the first time, the utility of the six-minute walk test (6MWT) in Chagas disease. The walked distance at 6MWT correlated negatively with the increased circulating levels of monocyte chemoattractant protein (MCP-1, r=-0.358, p=0.04) and natriuretic peptide type B (BNP, r=-0.349, p=0.04), as well as positively with ejection fraction deterioration (r=0.451, p=0.004), indicating that submaximal functional capacity of chagasic patients is related to the severity of the cardiopathy. 6MWT may constitute an auxiliary tool in the evaluation of the clinical status of Chagas disease patients. PMID- 17399823 TI - Thrombotic occlusion proximal to plaque rupture in acute myocardial infarction: evaluation by intravascular ultrasound and coronary angioscopy. AB - A 56-year-old man was admitted because of acute myocardial infarction for 10 h of onset. Emergent coronary angiography showed a total occlusion at the middle portion of the right coronary artery. After aspirating thrombus, high-grade stenosis was present in the distal segment. Angioscopy showed only a white thrombus, but not a yellow plaque nor a red thrombus at the initially occluded site. On the other hand, IVUS demonstrated large attenuation indicating a lipid core and cavity obstruction and angioscopy revealed a glittering yellow plaque and red thrombi in the distal segment. Precipitation of thrombus from the distal site to the middle site might have occurred. IVUS and angioscopy might be effective for detecting and evaluating the infarct-related vulnerable plaque and for making the therapeutic strategy when percutaneous coronary interventions are performed. PMID- 17399824 TI - Focale: study of systolic and diastolic heart failure in a French elderly population. AB - CONTEXT: The epidemic of heart failure (HF) in France has not been fully investigated and data on both the systolic and diastolic forms are limited. OBJECTIVES: To determine the prevalence, aetiology and treatments of both forms in French patients over 65 with HF (NYHA grade II-IV). PARTICIPANTS: Cross sectional study of 446 patients with HF recruited by 273 randomly selected cardiologists in France, with echocardiography and ECG available for central reading. MAIN MEASUREMENTS: Prevalence of diastolic (DHF) and systolic (SHF) HF using the left ventricular ejection fraction (LVEF). RESULTS: DHF, with a mean LVEF of 57.6+/-8.0%, was reported for 245 (54.9%) patients and SHF, with a mean LVEF of 33.33+/-8.0% was reported for 201 (45.1%) patients. Men were more prone to suffer SHF than DHF whereas for women the reverse was true. As compared to SHF, DHF endsystolic and enddiastolic volumes were smaller, the thickness/radius ratio was greater, with a longer E-wave deceleration time and a shorter peak filling rate. Aetiology and treatments were similar in both types of HF, except for ACE inhibitors and aldosterone antagonists, which were more frequently prescribed in SHF. CONCLUSION: This observational study provides further knowledge of SHF and DHF in outpatients. PMID- 17399825 TI - Demonstration of carotid plaque in Takayasu arteritis by multislice computed tomography. PMID- 17399826 TI - General aneurysmatosis due to cheese consumption: complications of an endocarditis caused by Lactococcus cremoris. AB - We describe a case of endocarditis due to Lactococcus cremoris associated with cheese consumption, that caused multiple mycotic aneurysms. Antibiotic treatment combined with surgical and radiological interventions resulted in full recovery. PMID- 17399827 TI - Current questions regarding the use of statins in patients with coronary heart disease. AB - The discovery of statins caused a revolution in the field of lipid intervention. Statins are drugs with a good safety profile. Their clinical benefit has been extensively documented in primary and secondary prevention of coronary heart disease. There is substantial evidence that the clinical outcome can be improved with aggressive statin treatment both in patients with unstable as well as with stable coronary heart disease. Also, early administration of statins in acute coronary syndromes is accompanied by rapid clinical benefits, mainly through their "pleiotropic" action (anti-inflammatory, anti-thrombotic, improvement of endothelial function, etc) which is probably a lipid-independent effect. Moreover, emerging data indicate that statins can achieve additional benefit when low density lipoprotein (LDL) cholesterol reduction is coupled with C-reactive protein reduction (<2 mg/L). The prevailing message from the recent statin trials is that intensive LDL cholesterol lowering treatment with statins achieves further clinical benefit beyond that achieved with standard statin therapy. This should encourage the medical community to consider prescribing statins in every coronary patient, aiming at LDL cholesterol levels <100 mg/dL, preferably in the range of 70-80 mg/dL in stable coronary patients, while in coronary patients at very high risk, the optional target for LDL cholesterol levels should be in the range of 50-70 mg/dL. PMID- 17399828 TI - Novel insoluble dye-labeled substrates for screening inulin-degrading microorganisms. AB - Inulin hydrogels were used to prepare insoluble colored substrates for screening of inulin-degrading microorganisms. Methycrylated inulin, synthesized with glycidyl methacrylate, was crossed-linked by free radical polymerization producing an insoluble hydrogel. The inulin hydrogels were colored with one of three covalent dyes (Reactive blue 4, Reactive orange 14, Reactive red 120). The efficacy of the colored inulin hydrogels was determined by comparing their performance to insoluble, colored amylose, xylan and HE-cellulose in screening assays. Novel substrates (alone or in combination with other insoluble colored substrates) were successfully used for screening inulin-degrading microorganisms from sludge samples. Selected strains were identified using 16S rDNA gene partial sequencing; all of them belong to the genus Bacillus. PMID- 17399829 TI - Data transformations in the analysis of community-level substrate utilization data from microplates. AB - BIOLOG EcoPlates provide one method for determination of functional diversity indices and community-level physiological profiling of microbial populations based on carbon substrate utilization. In this study, the effect of data transformation on BIOLOG EcoPlate data derived from wetland mesocosms and biofiltration systems was examined. Homoscedasticity, normality, and the number of linear correlations between variables were quantified and evaluated for data that had been transformed using either Taylor or logarithmic transforms. Subsequent multivariate analysis was implemented using the untransformed, Taylor transformed and logarithmic transformed data sets. The effect of data transformation and its effect on principle component analysis are presented. The transforms are shown to help increase homogeneity of variance, increase normality of the data, and increase the number of significant linear correlations for the data. Separate principle component analyses and ordinations of the data showed the transforms to be well suited to this type of data and in particular illustrate the ability of the logarithmic transform to reduce the influence of high leverage or outlying observations on the overall analysis and its robustness in terms of treating data from different ecological systems. Although BIOLOG EcoPlates were used in this study to illustrate the use of transformations on multivariate data, the techniques described may be employed on similar microplate data. In addition, if homoscedasticity, normality and the number of linear correlations within a data set are not evaluated and the possibility of transforming the data, using the Taylor, logarithmic or another transform, is not considered, erroneous analysis and misleading conclusions may be attained when performing multivariate analysis on microplate data. PMID- 17399830 TI - An evidence-based estimation of local control and survival benefit of radiotherapy for breast cancer. AB - BACKGROUND AND PURPOSE: Survival benefits from radiotherapy for breast cancer described in randomised trials represent only those patients eligible for trials. We estimated the benefit of radiotherapy as an adjuvant treatment for the entire population of breast cancer patients if evidence-based treatment guidelines were followed. MATERIALS AND METHODS: Evidences on 10-year local control and overall survival gain (radiotherapy vs no radiotherapy) were identified from review of literature. The data were incorporated into the optimal radiotherapy utilization tree that we previously reported for all categories of breast cancer patients and overall local control and survival benefits were estimated. RESULTS: The gains in 10-year local control and overall survival from optimal treatment of all breast cancer patients were 11.1% (95% CI 10.8-11.2%) and 3.1% (95% CI 3.0-3.4%), respectively. The stage-based estimates in local control and survival benefit were: 8% and 0% for Ductal Carcinoma in situ (DCIS), 12% and 2% for stage I-II cancers and 13% and 20% for stage III cancers. CONCLUSIONS: Our model was able to estimate the contribution of radiotherapy in breast cancer treatment if all patients were treated according to the recommended guidelines. These estimates could be used to benchmark population-based survival reports and to assess the cost-effectiveness of radiotherapy for breast cancer treatment. PMID- 17399831 TI - Expression of the mucus adhesion genes Mub and MapA, adhesion-like factor EF-Tu and bacteriocin gene plaA of Lactobacillus plantarum 423, monitored with real time PCR. AB - Expression of the mucus adhesion genes Mub and MapA, adhesion-like factor EF-Tu and bacteriocin gene plaA by Lactobacillus plantarum 423, grown in the presence of bile, pancreatin and at low pH, was studied by real-time PCR. Mub, MapA and EF Tu were up-regulated in the presence of mucus, proportional to increasing concentrations. Expression of MapA was up-regulated in the presence of 3.0 g/l bile and 3.0 g/l pancreatin at pH 6.5. Similar results were recorded in the presence of 10.0 g/l bile and 10.0 g/l pancreatin at pH 6.5. Expression of Mub was down-regulated in the presence of bile and pancreatin, whilst the expression of EF-Tu and plaA remained unchanged. Expression of Mub and MapA remained unchanged at pH 4.0, whilst expression of EF-Tu and plaA were up-regulated. Expression of MapA was down-regulated in the presence of 1.0 g/l l-cysteine HCl, suggesting that the gene is regulated by transcription attenuation that involves cysteine. PMID- 17399832 TI - Thermophilic Campylobacter spp. in salad vegetables in Malaysia. AB - The main aim of this study was to combine the techniques of most probable number (MPN) and polymerase chain reaction (PCR) for quantifying the prevalence and numbers of Campylobacter spp. in ulam, a popular Malaysian salad dish, from a traditional wet market and two modern supermarkets in Selangor, Malaysia. A total of 309 samples of raw vegetables which are used in ulam were examined in the study. The prevalences of campylobacters in raw vegetables were, for supermarket I, Campylobacter spp., 51.9%; Campylobacter jejuni, 40.7%; and Campylobacter coli, 35.2%: for supermarket II, Campylobacter spp., 67.7%; C. jejuni, 67.7%; and C. coli, 65.7%: and for the wet market, Campylobacter spp., 29.4%; C. jejuni, 25.5%; and C. coli, 22.6%. In addition Campylobacter fetus was detected in 1.9% of raw vegetables from supermarket I. The maximum numbers of Campylobacter spp. in raw vegetables from supermarkets and the wet market were >2400 and 460 MPN/g, respectively. PMID- 17399833 TI - Sorbitol co-feeding reduces metabolic burden caused by the overexpression of a Rhizopus oryzae lipase in Pichia pastoris. AB - To improve the specific production rate of Rhizopus oryzae lipase (ROL) in Pichia pastoris, a protein that triggers the unfolded protein response in P. pastoris, the effect of sorbitol/methanol mixed substrates was tested in batch and fed batch cultures. Remarkably, a different substrate consumption behaviour was observed depending on the host's phenotype (Mut(+) or Mut(s)) in batch cultures: when the methanol assimilation capacity is genetically reduced (Mut(s) phenotype), both substrates were consumed simultaneously, allowing not only a higher specific growth rate but also higher lipase levels (8.7-fold) compared to those obtained by cells growing on methanol as a sole carbon source in batch culture. This effect was not observed in Mut(+) phenotype, where the two substrates were consumed sequentially and the levels of heterologous product were only slightly higher (1.7-fold). A mixed substrate strategy was also applied to a Mut(s) fed-batch culture at a low methanol concentration set-point (0.5 gl(-1)). This resulted in a 2.2-fold increase in the heterologous protein level achieved, compared with the methanol-only feeding strategy. In addition, sorbitol co feeding permitted the achievement of higher specific growth rates, and avoided the drastic decrease of the specific production rate observed after the start of the induction phase when methanol was used as sole carbon source This resulted in a significant increase in the overall bioprocess volumetric productivity (2.2 fold) and specific productivity (1.7-fold). Moreover, whereas increased ROL gene dosage in Mut(s) strains have been previously reported to be deleterious for P. pastoris cells growing on methanol, sorbitol co-feeding allowed for sustained cell growth and lipase production. PMID- 17399834 TI - A new study of cell disruption to release recombinant thermostable enzyme from Escherichia coli by thermolysis. AB - Extraction of intracellular protein from Escherichia coli is traditionally achieved by mechanical, chemical or enzymatic disruption technology. In this study, a novel thermolysis method was used to disrupt E. coli cells to release a recombinant thermostable esterase. We found that heat treatment of E. coli was highly effective to destroy the integrity of bacterial cell walls and release the recombinant hyperthermophilic esterase at temperatures above 60 degrees C. The effects of temperature, pH and cell concentration on the efficiency of cell disruption were examined. The most effective temperature for cell disruption was at 80 degrees C. The pH and cell concentration had only minor effect on the release of the hyperthermophilic esterase. In addition, we found that the hyperthermophilic esterase could be purified at the early stage of the thermolysis, which is a major advantage of the thermolysis method. Finally, a comparison between thermolysis and traditional methods for the disruption of cells and the release of the thermostable enzyme was made. PMID- 17399835 TI - Feasibility of gas/solid carboligation: conversion of benzaldehyde to benzoin using thiamine diphosphate-dependent enzymes. AB - A carboligation was investigated for the first time as an enzymatic gas phase reaction, where benzaldehyde was converted to benzoin using thiamine diphosphate (ThDP)-dependent enzymes, namely benzaldehyde lyase (BAL) and benzoylformate decarboxylase (BFD). The biocatalyst was immobilized per deposition on non-porous support. Some limitations of the gas/solid biocatalysis are discussed based on this carboligation and it is also demonstrated that the solid/gas system is an interesting tool for more volatile products. PMID- 17399836 TI - Resistance of monocyte to HIV-1 infection is not due to uncoating defect. AB - Freshly isolated blood monocytes show an early postentry block to in vitro HIV-1 infection. Differentiation into monocyte-derived macrophages (MDMs) is required to allow HIV replication in this cell type. In this study, we investigated whether the resistance of monocyte to HIV infection stemmed from uncoating defect. Monocyte and MDM lysates induced HIV-1 core uncoating to a comparable degree. This suggests that monocyte lacks a factor(s) essential for viral reverse transcription and/or contains a factor(s) interfering with this step. PMID- 17399837 TI - RNA interference against hepatitis B virus with endoribonuclease-prepared siRNA despite of the target sequence variations. AB - RNA interference (RNAi) has proven to be very powerful in inhibiting hepatitis B virus (HBV) replication by cell culture and mouse model studies. We have previously reported that endoribonuclease-prepared short interfering RNAs (esiRNAs) were able to inhibit HBV replication more efficiently than synthesized siRNAs. Here we tested the hypothesis that esiRNAs are able to inhibit gene expression with limited mutations within the target region. Target sequences with different similarities to esiHBVP (esiRNA targeting the DNA polymerase and S antigen of Hepatitis B virus) were amplified and cloned into the 3' untranslated region of HBsAg, respectively. When the obtained expression vectors were co transfected with esiHBVP into CHO cells, HBsAg expression was suppressed with same efficiency regardless of the target sequence similarities. In HepG2 cells, esiHP9 based on one of the amplified sequence that sharing 87% similarity to the target region suppressed HBsAg expression effectively and dose dependently. In vivo experiment showed that a single dose of 5 microg esiHP9 was able to reduce HBsAg and HBeAg level in the mouse sera by 88 and 77% despite of its 87% similarity to the target sequence, which was as good as esiHBVP that is 100% similar to the target sequence. All the data suggest that esiRNA can tolerate limited target sequence variations without losing its inhibitory capacity. It would be very helpful to suppress virus replication by RNAi despite of their high mutation rate. PMID- 17399838 TI - Anti-PEG IgM elicited by injection of liposomes is involved in the enhanced blood clearance of a subsequent dose of PEGylated liposomes. AB - Earlier we reported that PEGylated liposomes lose their long-circulating characteristic when they are administrated twice in the same animal with certain intervals (referred to as the accelerated blood clearance (ABC) phenomenon). We proposed that anti-PEG IgM, induced by the PEGylated liposomes, is responsible for the phenomenon, based on the observation that IgM thus produced selectively binds to the surface of PEGylated liposomes, subsequently leading to substantial complement activation. Interestingly, we found that under certain circumstances administration of conventional liposomes without PEG-coating also caused a strong ABC response upon injection of a second dose of PEGylated liposomes, but not of conventional liposomes. This suggests that also conventional liposomes not modified with PEG can promote an IgM response against PEG. We report here that, irrespective of the presence or absence PEG-coating, a single first dose of liposomes is capable of inducing a strong anti-PEG IgM response and, under certain circumstances, also weak responses against other lipid components. A good correspondence was observed between the amount of IgM associating with both PEGylated and conventional liposomes, concomitant complement activation triggered by those liposomes and the magnitude of the ABC phenomenon against those liposomes. Taken together, our results demonstrate that the ABC phenomenon is fully attributable to production of anti-PEG IgM by the first dose of liposomes and the subsequent complement activation upon a second dose of PEGylated but not conventional liposomes. Although the responsible immunogenic epitopes of the liposomes remain to be determined, the immunogenicity of 'empty' liposomes presents a serious concern in the development of liposomal formulations and their use in the clinic. Furthermore, our findings as described here raise important concerns with regard to the safety and efficiency of liposomes currently under development for clinical use. PMID- 17399839 TI - PLGA nanoparticles for oral delivery of cyclosporine: nephrotoxicity and pharmacokinetic studies in comparison to Sandimmune Neoral. AB - The cyclosporine-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) were prepared by the emulsion-diffusion-evaporation method and were optimized for particle size and entrapment efficiency. The optimized particles were 143.3+/-8.7 nm in size with narrow size distribution and 71.9+/-1.7% entrapment efficiency at 20% w/w initial drug loading when prepared with 0.1% w/v of Didodecylmethylammonium bromide (DMAB) as stabilizer. These particulate carriers exhibited controlled in vitro release of cyclosporine for 23 days at a nearly constant rate and showed very good hemocompatibility in vitro. The nanoparticulate formulation showed significantly higher intestinal uptake as compared to Sandimmune Neoral and cyclosporine suspension. The relative bioavailability of nanoparticulate formulation was found to be 119.2% as compared to Sandimmune Neoral. A marked difference in the pharmacokinetic profile between nanoparticulate and Sandimmune Neoral formulations was observed where nanoparticulate formulation showed controlled release of cyclosporine over 5 days, on the other hand, the marketed formulation showed a sharp Cmax with a 3 day release profile. The nanoparticulate formulation exerted significantly lower nephrotoxicity in the rats as compared to Sandimmune Neoral, which was evidenced by lower blood urea nitrogen (BUN), plasma creatinine (PC) and malondialdehyde (MDA) levels in plasma and kidney. The results were further supported by the histopathological changes in kidneys. Together, these results indicate that PLGA NPs have greater potential for oral delivery of cyclosporine. PMID- 17399840 TI - [Benefit of isokinetic evaluations of knee before and after anterior cruciate ligament reconstruction in soccer players]. AB - AIM: The aim of this study was to analyse the evolution of the isokinetic performance of the knee flexor and extensor muscles in soccer players before and after intra-articular anterior cruciate ligament reconstruction (Kennet-Jones). METHODS: Two isokinetic evaluations were carried out before surgery and after rehabilitation (i.e. 4 months later) in 18 soccer players. RESULTS: The initial evaluation showed that the stabilizing muscles of the knee were affected differently after lesion of the external anterior crossed ligament. At the opposite of the flexor muscles, the performance of the knee extensor muscles of the injured leg was significantly reduced as compared with that of the healthy leg (peak torque at 90 degrees /s, -16%; power at 180 degrees /s, -14%; total work at 240 degrees /s, -11%). Even if 4 months after surgery, this deficit was accentuated (peak torque at 90 degrees /s, -26%; power at 180 degrees /s, -23%; total work at 240 degrees /s, -19%), the preoperative results of the knee extensor muscle do not condition the postoperative performance. CONCLUSION: After a rupture of the external anterior crossed ligament, a regular program of isokinetic evaluation of the knee seems to be relevant in the follow-up of the wounded athlete. The evaluation before surgery can be used as reference, and from a psychological point of view, this evaluation can create confidence in the athlete during hospitalization. A second evaluation 4 months after surgery can be used to quantify the muscular deficit to direct the exercises of rehabilitation. PMID- 17399841 TI - Physical activity in the management of cancer-related fatigue induced by oncological treatments. AB - OBJECTIVE: We aimed to review the literature on physical activity in the management of cancer-related fatigue. MATERIALS AND METHODS: A search of PubMed for English- or French-language articles using the keywords fatigue, cancer, exercise, physical activity, sport, randomized, and controlled. RESULTS: We found reports of 11 randomized or controlled studies, all of them in English. Most involved small numbers of patients. Four of the studies reported that physical exercise had an effect on the fatigue: three a positive effect and one a negative effect. One of these four studies was carried out in patients still undergoing treatment (hormone therapy). Of the three post-treatment studies, the two finding positive effect involved breast cancer patients quite a while after the end of treatment (chemotherapy and/or radiotherapy), whereas the study finding a negative effect involved patients 1 month after treatment, all with chemotherapy. CONCLUSION: Physical activity appears to be a good way of combating fatigue induced by various cancer therapies. However, many questions remain, which highlights the need for randomized studies with sufficient numbers of patients. PMID- 17399842 TI - Ethnicity, substance use, and response to booster hepatitis B vaccination in anti HBs-seronegative adolescents who had received primary infantile vaccination. AB - BACKGROUND/AIMS: In this revaccination study, we explored the determinants of response to booster hepatitis B (HB) vaccination in anti-HBs-seronegative adolescents who had received primary HB vaccination 15-18 years before. RESULTS: After controlling for prebooster anti-HBs levels, cigarette smoking, betel-quid chewing, alcohol drinking, and indigenous ethnicity were significantly associated with elevated risks of non-response to booster HB vaccination. The adjusted odds ratios (aORs) were 3.21 (CI: 1.33-7.84), 8.78 (CI: 2.03-37.94), 2.64 (CI: 1.15 6.02), and 2.46 (CI: 1.28-4.72), respectively. Among adolescents with undetectable prebooster anti-HBs titers, only indigenous ethnicity significantly associated with elevated risk, with an adjusted OR of 2.57 (CI: 1.20-5.54), of non-response to booster HB vaccination. On the contrary, the influences of cigarette smoking, betel-quid chewing, and alcohol drinking were restricted to adolescents with prebooster anti-HBs titers of 0.1-9.9mIU/mL. The corresponding multivariate-adjusted ORs were 5.70, 17.41, and 3.72, respectively. Adolescents who smoked cigarettes and chewed betel-quid were at highest risk of non-response (aOR, 25.3; CI: 2.97-215.7). CONCLUSIONS: A booster dose of HB vaccine may be insufficient to induce immunological response in healthy adolescents who had undetectable prebooster anti-HBs titers or who were of Malay-Polynesian ethnicity. Responses to booster vaccination are probably modified by recent cigarette smoking and/or betel-quid chewing. PMID- 17399843 TI - In vivo altered unfolded protein response and apoptosis in livers from lipopolysaccharide-challenged cirrhotic rats. AB - BACKGROUND/AIMS: Endoplasmic reticulum (ER)-related unfolded protein response (UPR) is mediated by PKR-like ER kinase (PERK), ATF6 and IRE1. PERK phosphorylates eukaryotic translation initiation factor-2alpha (eIF2alpha) to attenuate protein synthesis, including in NF-kappaB-dependent antiapoptotic proteins. We hypothesized that an altered UPR in the liver may sensitize cirrhotic livers to LPS-induced, TNFalpha-mediated apoptosis. Thus, we examined in vivo UPR and NF-kappaB activity in livers from cirrhotic and normal LPS challenged rats. METHODS: Livers were harvested in rats that did or did not receive LPS. RESULTS: Under baseline conditions, no UPR was found in normal livers while PERK/eIF2alpha and ATF6 pathways were activated in cirrhotic livers. After LPS, in normal livers, the PERK/eIF2alpha pathway was transiently activated. ATF6 and IRE1 were activated. In cirrhotic livers, the PERK/eIF2alpha pathway remained elevated. ATF6 and IRE1 pathways were altered. LPS-induced, NF kappaB-dependent antiapoptotic proteins increased in normal livers whereas their expression was blunted at the posttranscriptional level in cirrhotic livers. CONCLUSIONS: Cirrhotic livers exhibit partial UPR activation in the basal state and full UPR, although altered, after LPS challenge. Sustained eIF2alpha phosphorylation, a hallmark of cirrhotic liver UPR, is associated with a lack of LPS-induced accumulation of NF-kappaB-dependent antiapoptotic proteins which may sensitize cirrhotic livers to LPS/TNFalpha-mediated apoptosis. PMID- 17399844 TI - Expression of the subgenomic hepatitis C virus replicon alters iron homeostasis in Huh7 cells. AB - BACKGROUND/AIMS: Infection with hepatitis C virus (HCV) is associated with alterations in body iron homeostasis by poorly defined mechanisms. To seek for molecular links, we employed an established cell culture model for viral replication, and assessed how the expression of an HCV subgenomic replicon affects iron metabolism in host Huh7 hepatoma cells. METHODS: The expression of iron metabolism genes and parameters defining the cellular iron status were analyzed and compared between parent and replicon Huh7 cells. RESULTS: By using the IronChip microarray platform, we observed replicon-induced changes in expression profiles of iron metabolism genes. Notably, ceruloplasmin mRNA and protein expression were decreased in replicon cells. In addition, transferrin receptor 1 (TfR1) was also downregulated, while ferroportin levels were elevated, resulting in reduced iron uptake and increased iron release capacity of replicon cells. These responses were associated with an iron-deficient phenotype, manifested in decreased levels of the "labile iron pool" and concomitant induction of IRE-binding activity and IRP2 expression. Furthermore, hemin-treated replicon cells exhibited a defect in retaining iron. The clearance of the replicon by prolonged treatment with interferon-alpha only partially reversed the iron-deficient phenotype but almost completely restored the capacity of cured cells to retain iron. CONCLUSIONS: We propose that Huh7 cells undergo genetic reprogramming to permit subgenomic viral replication that results in reduction of intracellular iron levels. This response may provide a mechanism to bypass iron mediated inactivation of the viral RNA polymerase NS5B. PMID- 17399845 TI - Are there ethnicity-based differences in the evaluation of individuals with abnormal liver biochemistries? AB - BACKGROUND/AIMS: Recent studies suggested NAFLD is less infrequent in African Americans (AA) than in Caucasians but it is unclear if this difference is biological or due to under-recognition/under-referral. This study examined if there is an ethnicity-based difference in obtaining liver biochemistries or evaluating abnormal liver biochemistries by primary care physicians. METHODS: This study consisted of 45,016 AA and 49,660 Caucasians seen at our primary care clinics over a 3-year period. From these two groups, we identified patients with elevated aminotransferases (AA: 3676, Caucasians: 4644) and elevated bilirubin (AA: 1295, Caucasians: 1199) based on predefined criteria. Subsequently, we assessed the proportion of patients in each group who had liver-specific evaluation (viral serologies, abdominal imaging or GI clinic visit). RESULTS: Among patients with elevated aminotransferases, compared to Caucasians, AA did not have lower testing for viral hepatitis (26% vs. 25%), imaging (16% vs. 13%) or GI clinic visits (17% vs. 17%). Similarly, we did not observe clinically significant difference in the evaluation of elevated bilirubin between AA and Caucasians (viral serologies: 22% vs. 22%; imaging: 25% vs. 27%; GI clinic: 15% vs. 21%). CONCLUSIONS: Under-recognition and under-referral are not likely to explain the reported ethnic differences in the prevalence of NAFLD. PMID- 17399846 TI - Depression in patients with primary biliary cirrhosis and primary sclerosing cholangitis. AB - BACKGROUND/AIMS: Former studies reported a high prevalence of depression in patients with primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). These studies hypothesized that the presence of depression could explain the fatigue experienced by these patients. METHODS: Our aim was to study the prevalence of depression in a Dutch population with PBC and PSC. In addition, to investigating the effects of using an additional diagnostic structured psychiatric interview, after screening with the Beck Depression Inventory (BDI), a self-report severity scale instrument used in former studies. Patients with PBC and PSC (n=92) completed the BDI. Patients with scores of 10 or higher (n=39) were interviewed using a structured psychiatric interview. Patients with scores lower than 10 were at random (30/53, 57%) also interviewed using a structured psychiatric interview. RESULTS: Of the 92 patients that were included 42% had depressive symptoms according to the BDI. However, of these patients only 3.7% had a depressive syndrome according to the DSM-IV criteria as assessed with the structured psychiatric interview. CONCLUSIONS: The prevalence of a depressive disorder in patients with PBC and PSC is not higher than in the general population. Fatigue in patients with PBC and PSC cannot be explained by depression. PMID- 17399847 TI - Prevention of hepatocarcinogenesis with phosphatidylcholine and menaquinone-4: in vitro and in vivo experiments. AB - BACKGROUND/AIMS: We examined whether phosphatidylcholine inhibited growth of hepatic cancer, as previously shown for menaquinone-4 (vitamin K2). METHODS: Growth inhibitions by phosphatidylcholine and/or menaquinone-4 and apoptosis induction by phosphatidylcholine were evaluated in vitro using human hepatic cancer cell lines (Hep-3B, Hep-G2, HuH-7, and Alexander). Effects of these agents were then investigated in male Sprague-Dawley rats against hepatocarcinogenesis induced by diethylnitrosamine plus phenobarbital. All rats were killed to examine livers to evaluate inhibitory potential macroscopically and immunohistochemically using an antibody against the marker of carcinogenesis, glutathione S-transferase and apoptotic induction by phosphatidylcholine using TUNEL staining. Blood samples were obtained by cardiac puncture. RESULTS: In vitro, phosphatidylcholine and menaquinone-4 each inhibited cancer cell growth and phosphatidylcholine induced apoptosis dose-dependently. Moreover, exposure to both synergistically inhibited growth in Hep-3B. In vivo, diets containing phosphatidylcholine with or without menaquinone-4 significantly reduced the number of macroscopic hepatic tumor nodules and the extent of abnormally immunoreactive foci conserving hepatic function on serum examinations compared with controls given only the carcinogens. Moreover, phosphatidylcholine supplementation induced apoptosis on TUNEL staining of liver sections. CONCLUSIONS: Given together, phosphatidylcholine and menaquinone-4 may exhibit synergy against hepatocarcinogenesis conserving hepatic function that could benefit patients at high risk for hepatocellular carcinoma. PMID- 17399848 TI - Psoriasis patients with diabetes type 2 are at high risk of developing liver fibrosis during methotrexate treatment. AB - BACKGROUND/AIMS: We investigated the impact of diabetes mellitus type 2, overweight, alcohol over-consumption, and chronic hepatitis B or C as risk factors, for liver fibrosis in psoriasis patients treated with methotrexate. METHODS: One hundred and sixty-nine liver biopsies from 71 patients who underwent liver biopsies as part of the monitoring of methotrexate treatment for psoriasis were reviewed. Fibrosis, steatosis and inflammation were staged according to the NAFLD activity score. RESULTS: Twenty-six patients had one or more of the risk factors and 25 (96%) of these (median cumulative dose methotrexate 1500 mg) developed liver fibrosis. Of those without risk factor, 26 (58%) (p=0.012) developed fibrosis (median cumulative dose methotrexate 2100 mg). Ten (38%) of the patients with risk factor(s) had severe fibrosis (stage 3-4) (mean cumulative dose methotrexate 1600 mg), while four (9%) (p=0.0012) of those without risk factors had severe fibrosis (median cumulative dose methotrexate 1900 mg). CONCLUSIONS: Patients with methotrexate treated psoriasis and risk factors for liver disease, especially diabetes type 2 or overweight, are at higher risk of developing severe liver fibrosis compared to those without such risk factors, even when lower cumulative methotrexate doses are given. PMID- 17399849 TI - Achieving Millennium Development Goals for health: building understanding, trust and capacity to respond. AB - Biomedical interventions promise achievement of health-related Millennium Development Goals provided social-, capacity- and knowledge-based constraints to scaling up and reaching marginalized people at risk, are addressed, and balance between prevention and treatment is struck. We argue for a new approach: multi stakeholder capacity building and learning for empowerment: MuSCLE. MuSCLE is used as a way to frame three systemic weaknesses in traditional health science and policy approaches: (1) a lack of engagement with people at risk to build a collective understanding of the contexts of health problems, including social drivers; (2) a lack of multi-criteria evaluation of alternative interventions; (3) a lack of attention paid to integrated capacity building. The MuSCLE framework responds in three ways: (1) participatory assessment of the ecological, socio-cultural, economic and political contexts of health, identifying priorities using risk and vulnerability science, and modeling drivers; (2) selection among intervention alternatives that makes ecological, socio-cultural, economic and political tradeoffs transparent; (3) integrated capacity building for sustainable and adaptive interventions. Literature and field lessons support the argument, and guidelines are set down. A MuSCLE approach argues for a transformation in health science and policy in order to achieve Millennium Development Goals for health. PMID- 17399850 TI - A multi-center phase II study of weekly topotecan as second-line therapy for small cell lung cancer. AB - PURPOSE: To determine the response rate, toxicity, failure free and overall survival of weekly topotecan in patients with relapsed small cell lung cancer who received one prior platinum based chemotherapy. PATIENTS AND METHODS: Twenty two patients with relapsed disease after response to one prior chemotherapy with or without radiotherapy and patients with relapse more than 90 days after their last therapy received topotecan 4mg/m(2) intravenous over 30min on days 1,8,15; every 4 weeks (3 weeks on and 1 weeks off). Chemotherapy was given until disease progression or unacceptable toxicity. RESULTS: Of 22 patients, none of the patients responded to weekly topotecan therapy. Four patients had stable disease. After a median follow up of 1 year, median time to progression was 6 weeks and median survival was 5 months. The common toxicities associated with this regimen were anemia, thrombocytopenia, fatigue, GI side effects and alopecia. CONCLUSION: Weekly topotecan was well tolerated but ineffective in this trial. Although commonly used, weekly regimen of topotecan should be used with caution in relapsed SCLC. PMID- 17399851 TI - Relationships between body size and abundance in ecology. AB - Body size is perhaps the most fundamental property of an organism and is related to many biological traits, including abundance. The relationship between abundance and body size has been extensively studied in an attempt to quantify the form of the relationship and to understand the processes that generate it. However, progress has been impeded by the under appreciated fact that there are four distinct, but interrelated, relationships between size and abundance that are often confused in the literature. Here, we review and distinguish between these four patterns, and discuss the linkages between them. We argue that a synthetic understanding of size-abundance relationships will result from more detailed analyses of individual patterns and from careful consideration of how and why the patterns are related. PMID- 17399852 TI - A construct analysis of meal convenience applied to military foods. AB - The present research investigates the concept of food convenience within the institutional framework of military feeding. The approach views food-related convenience in terms of two broad dimensions: "type of convenience" and "timing of convenience." A discrete choice experiment was conducted with US military personnel (n=179) regarding their perceptions of the (in)convenience associated with the use and consumption of low-preparation, all-in-one, military meals (MREs meals, ready-to-eat). The obtained data strongly suggest that perceived (in)convenience, time and effort are separate constructs. A food provisioning process perspective was captured in the "timing of convenience" dimension, and the contribution of different stages in the consumption process to the perceived convenience of the meal situation was empirically demonstrated. The latter result has important implications for the study of food convenience outside this specific population and context. As opposed to the product perspective that is currently predominant in the literature, it demonstrates the necessity of adopting a meal perspective in analysing food-related convenience. PMID- 17399853 TI - Early impairment of synaptic plasticity in patients with Down's syndrome. AB - We investigated synaptic plasticity in persons with Down' syndrome (DS) and control subjects used paired associative stimulation (PAS) protocol, a paradigm capable of producing long-term potentiation (LTP)-like changes in the sensorimotor system. After PAS, patients showed less LTP-like plasticity compared to control subjects. Abnormal motor cortex synaptic plasticity may play a role in the development of motor signs in DS. PMID- 17399854 TI - Expression of GDNF transgene in astrocytes improves cognitive deficits in aged rats. AB - Glial cell line-derived neurotrophic factor (GDNF) was assayed for its neurotrophic effects against the neuronal atrophy that causes cognitive deficits in old age. Aged Fisher 344 rats with impairment in the Morris water maze received intrahippocampal injections at the dorsal CA1 area of either a lentiviral vector encoding human GDNF or the same vector encoding human green fluorescent protein as a control. Recombinant lentiviral vectors constructed with human cytomegalovirus promotor and pseudotyped with lyssavirus Mokola glycoprotein specifically transduced the astrocytes in vivo. Astrocyte-secreted GDNF enhanced neuron function as shown by local increases in synthesis of the neurotransmitters acetylcholine, dopamine and serotonin. This neurotrophic effect led to cognitive improvement of the rats as early as 2 weeks after gene transduction. Spatial learning and memory testing showed a significant gain in cognitive abilities due to GDNF exposure, whereas control-transduced rats kept their performance at the chance level. These results confirm the broad spectrum of the neurotrophic action of GDNF and open new gene therapy possibilities for reducing age-related neurodegeneration. PMID- 17399855 TI - [Pregnancy and thyroid disorders]. AB - Association between pregnancy and thyroid disorders is a frequent event. In case of maternal hyperthyroidism, strict guidelines relying mainly on foetal thyroid monitoring echographic scanning will allow in most cases the delivery of a healthy euthyroid newborn. Hypothyroidia, providing an adequate substitution has no significant impact on pregnancy, biological monitoring is the key of monitoring. Iodine deficiency is a matter of concern when considering neurodevelopmental outcome, however it is still an unsolved issue in France. However involvement of a multidisciplinary team is of good practice in most of the cases. PMID- 17399856 TI - [Quality of life and scales measuring]. AB - SCOPE: To present a review of the concept and the measurement of quality of life. CURRENT SITUATION AND SALIENT POINTS: The concept of quality of life was developed in the seventies aiming to complete the medical evaluation. It appeared with improvement of disease prognosis, medical evaluation and the development of patient autonomy of patients. The methodology used to construct quality of life scales is presented. Problems raised by adaptation of instruments for use in other culture are briefly discussed. PERSPECTIVES: These scales are now used in clinical research, medical economic studies and day-to-day practice. However, ethical issues surrounding these measures persist. PMID- 17399857 TI - Peeling back the layers of learning: a classroom model for problem-based learning. AB - This paper aims to provide an informative discussion with underpinning rationales about the use of a problem-based learning (PBL) classroom model, supported by a structured process for undertaking PBL. PBL was implemented as a main teaching and learning strategy for a diploma in nursing programme as advised by the Department of Health [Department of Health., 1999. Making a difference: Strengthening the Nursing, Midwifery and Health Visiting Contribution to Health and Health Care. Department of Health, London.] and the United Kingdom Central Council for nurses, midwifes and health visitors [United Kingdom Central Council for Nursing, Midwifery and Health Visiting, 1999. Fitness for Practice. UKCC, London.]. The implementation and change to the PBL approach is not without challenges, and so it was considered important to facilitate this change effectively. Through ongoing reflection, peer discussions and continuous review of the literature following studies at Masters Level, it was identified that the design of a model may guide students and facilitators who were new to the PBL process to help students identify relevant learning needs and thus enable them to achieve the learning outcomes of a dynamic curriculum [Darvill, A., 2000. Developing Problem-based Learning in the Nursing Education Curriculum: A Case Study. Unpublished MSc Dissertation, University of Huddersfield, Huddersfield; McLoughlin, M., 2002. An Exploration of the Role of the Problem-based Learning Facilitator: An Ethnographic Study of Role Transition in a Higher Education Institution 'Paradigm Shift or New Ways of Working'. Unpublished MSc Dissertation. University of Huddersfield, Huddersfield.]. In this paper the key components of the model will be described. PMID- 17399858 TI - Quality assessment and comparison of grading between examiners and supervisors of Bachelor theses in nursing education. AB - This study compares supervisors' and examiners' grading of quality of theses at Bachelor level in nursing. An instrument developed to asses the quality of theses was used. Eight aspects of quality were rated. One hundred and fifteen theses were rated by both examiner and supervisor. Significant correlations were found between examiners' and supervisors' ratings of all aspects of quality. Good agreement was found in 89-96% of the ratings on individual aspects of quality. The means of differences between ratings were small but significantly differed from zero in four out of eight aspects. In theses rated low for quality of language and formality, differences between examiners' and supervisors' ratings on all aspects of quality were significantly larger than in theses rated high for quality of language and formality. The general conclusion is that the evaluations made by examiners and by supervisors corresponded well. Differences found indicates that examiners in general give lower scores than supervisors especially on quality aspects that are most closely related to research methods and experiences. This study is part of a larger project that is investigating quality of theses and the student's attitudes and learning experiences of writing theses. PMID- 17399859 TI - A new highly effective anticysticercosis vaccine expressed in transgenic papaya. AB - The use of transgenic plants as new antigen-delivery systems for subunit vaccines has been increasingly explored. We herein report progress toward a papaya-based vaccine against cysticercosis. Synthetic peptides (KETc1, KETc12, KETc7) were successfully expressed in 19 different transgenic papaya clones and found to be immunogenic. Complete protection against cysticercosis was induced with the soluble extract of the clones that expressed the higher levels of transcripts in up to 90% of the immunized mice. This study represents a key step towards the development of a more effective, sustainable and affordable oral subunit vaccine against human and pig cysticercosis. PMID- 17399860 TI - The PGRS domain of Mycobacterium tuberculosis PE_PGRS Rv1759c antigen is an efficient subunit vaccine to prevent reactivation in a murine model of chronic tuberculosis. AB - A third of the world population is latently infected with Mycobacterium tuberculosis and many cases of active tuberculosis arise from latent bacilli reactivation. Thus, it is important to design new vaccines to prevent reactivation. Using an experimental model of chronic tuberculosis in B6D2F1 mice, we observed constant expression of Rv1759c antigen, a member of the PE_PGRS gene family, on the cell wall of phagocytosed mycobacteria by activated macrophages located in lung granulomas. This antigen induced production of IFN-gamma after stimulation of cell suspensions from mediastinal lymph nodes. It was notorius that chronic infected mice immunized with this antigen and treated with corticosterone to induce reactivation showed not change in colony forming units (CFU), compared with the significant bacilli increase in non-vaccinated mice treated with corticosterone. These results suggest that this antigen could play an important role in the immune response that maintains latent infection, and could therefore, be a good candidate as a new subunit vaccine to prevent disease reactivation. PMID- 17399862 TI - The threat of peste des petits ruminants: progress in vaccine development for disease control. AB - Peste des petits ruminants (PPR) is a highly contagious animal disease caused by a virus in the genus Morbillivirus, family Paramyxoviridae. This infection is responsible for high morbidity and mortality in sheep and goats and in some small wild ruminant species. The huge number of small ruminants, which are reared in the endemic areas makes PPR a serious disease threatening the livelihood of poor farmers. Taking advantage of the closely relationship between rinderpest and PPR viruses, the attenuated rinderpest vaccine was used in the control of PPR. It is now replaced by the homologous attenuated PPR vaccine. Unfortunately, animals that have received this vaccine cannot be distinguished serologically from infected animals. With the advent of DNA recombinant technology, efforts are being made to develop effective PPR marker vaccines to enable such differentiation and which would allow countries to implement both vaccination and disease surveillance programmes at the same time. PMID- 17399861 TI - Improving tumor targeting and therapeutic potential of Salmonella VNP20009 by displaying cell surface CEA-specific antibodies. AB - Genetically modified Salmonella typhimurium VNP20009 (VNP) is a useful vehicle for cancer therapy and vaccine development but exhibits limited tumor targeting in vivo. We engineered a novel VNP derivative that expressed carcinoembryonic antigen (CEA)-specific single chain antibody fragments (scFv) on the cell surface to increase tumor-specific targeting. There was significant scFv cell surface display visualized by flow cytometry and confocal microscopy when cells were probed with fluorescently labeled CEA. Atomic force microscopy (AFM) measurements on whole bacteria confirmed binding of unlabeled CEA to the displayed scFv. The modified VNP strain exhibited increased localization in the upper gastrointestinal tract of CEA transgenic mice and accumulated in CEA-expressing tumors. Furthermore, treatment with a single dose of the VNP derivative inhibited growth of MC38CEA tumors and was associated with local accumulation of CD3(+) T cells and CD11b(+) macrophages. The display of antibody fragments on the surface of VNP represents a novel strategy for both targeting CEA-expressing tumors and increasing the immunogenicity of Salmonella-based vaccines for cancer. PMID- 17399863 TI - Radiochlorine concentration ratios for agricultural plants in various soil conditions. AB - Long-term field experiments have been carried out in the Chernobyl exclusion zone in order to determine the parameters governing radiochlorine ((36)Cl) transfer to plants from four types of soil, namely, Podzoluvisol, Greyzem, Phaeozem and Chernozem. Radiochlorine concentration ratios (CR=concentration of (36)Cl in the fresh plant material divided by its concentration in the dried soil in the upper 20 cm layer) were obtained in green peas (2.6+/-0.4), onions (1.5+/-0.5), potatoes (8+/-1), clover (90+/-26) and ryegrass (158+/-88) hay, oat seeds (36+/ 23) and straw (305+/-159), wheat seeds (35+/-10) and straw (222+/-82). These values correlate with the stable chlorine values for the same plants. It was shown that (36)Cl plant/soil CR in radish roots (CR=9.7+/-1.4) does not depend on the stable chlorine content in the soil (up to 150 mgkg(-1)), soil type and thus, that stable chlorine CR values (9.4+/-1.2) can also be used for (36)Cl. Injection of additional quantities of stable chlorine into the soil (100 mgkg(-1) of dry soil) with fertilizer does not change the soil-to-plant transfer of (36)Cl. The results from a batch experiment showed that chlorine is retained in the investigated soils only by live biota and transfers quickly (in just a few hours) into the soil solution from dry vegetation even without decomposition of dead plants and is integrated in the migration processes in soil. PMID- 17399864 TI - Development of an evidence-based domestic violence guideline: supporting perinatal women-centred care in Japan. AB - OBJECTIVE: to develop an evidenced-based, women-centred care clinical guideline designed to assist midwives and other health-care providers in Japanese hospitals, clinics and midwifery offices, in identifying and supporting potential or actual perinatal victims of domestic violence. DESIGN: systematic review and critical appraisal of extant research; structured assessment of clinical guideline development. METHOD: systematic and comprehensive literature search. Appraisal of Guidelines for Research and Evaluation (AGREE) was used to assess the guideline development for purposes of assuring methodological quality. FINDINGS: electronic searches of medical and nursing databases between February and December 2003 retrieved 2392 articles. Selected as evidence were 157 articles yielding 28 recommendations aligned to clinical assessment questions. KEY CONCLUSIONS: using expert consensus and external reviews, recommendations were generated that provided the at-risk perinatal group with the best possible practice available to prevent further harm. IMPLICATIONS FOR PRACTICE: the evidenced-based clinical guideline fosters a supportive environment for educating health-care providers on domestic violence, and to improve clinic access for at risk perinatal women. Information on domestic violence and a negotiated midwife client safety plan can be initiated for potential or actual victims of domestic violence, and is achieved through understanding the risks of the woman and her fetus or baby, while respecting the woman's intention. PMID- 17399865 TI - Re: The effects of scaphoid and colles casts on hand function. PMID- 17399867 TI - Re: Two significant complications following percutaneous needle fasciotomy in a patient on anticoagulants. PMID- 17399869 TI - Scanning electron microscopic findings of the gliding surface of the A1 pulley in trigger fingers and thumbs. AB - The gliding surface of the A1 pulley was studied in 20 cases of primary trigger finger by scanning and transmission electron microscope. In 12 normal specimens, the whole deep surface was covered uniformly by an amorphous extracellular matrix. In the pathological samples, there was the same general surface appearance but, also, areas, varying in shape and dimension where loss of the extracellular matrix had exposed the collagen fibres and a few cells of the middle layer of the pulley. There were also changes typical of "chondroid metaplasia". These data were confirmed by transmission electron microscopy. The fragmented areas are probably the result of altered forces of friction between the pulley and the flexor tendon and may be the "gate" through which the forces of friction cause chondroid-metaplasia in the underlying fibrous tissue, a phenomenon recognised to be one step in the pathogenesis of trigger finger. PMID- 17399870 TI - The surgical treatment and outcomes of high-pressure injection injuries of the hand. AB - High-pressure injection injuries to the hand are uncommon soft tissue traumas of the hand which are frequently underestimated. They are, however, not uncommon among workers in industry using paint, automotive grease, solvents and diesel oil. The prognostic factors are the type, amount and temperature of the material and the pressure of injection. The results of 14 surgically treated high-pressure injection injuries of the hand with a minimum of two years follow-up are reported. Ten cases required soft tissue debridement and foreign body removal. Six required reconstructive microsurgical procedures and one underwent digital tip amputation, which followed major and devastating soft tissue problems. In the present study, we describe the mechanism of the soft tissue damage and the principles of treatment. This study confirms the fact that high-pressure injection injury to the hand is a significant problem, which can easily lead to serious sequelae and, even, amputation. PMID- 17399871 TI - To suspend or not to suspend: a randomised single blind trial of simple trapeziectomy versus trapeziectomy and flexor carpi radialis suspension. AB - The thumb carpometacarpal joint is the second most common site of osteoarthritis in humans. There are numerous operations for the condition but, perhaps, the commonest is trapeziectomy, sometimes supplemented by a suspension procedure, most commonly using part of the flexor carpi radialis tendon. In order to determine whether there is an advantage to a suspension procedure, or not, 65 patients with Eaton and Glickel Grade III or IV arthritis of the carpometacarpal joint of their thumbs were randomised into either undergoing trapeziectomy alone (with no wiring) or a trapeziectomy with flexor carpi radialis suspension. Patient satisfaction from both operations was similar. There was increased range of movement in the trapeziectomy alone group, but there was no difference in grip or pinch strength. Measurement of the gap on X-ray left by the trapeziectomy was less when trapeziectomy alone was performed. PMID- 17399872 TI - Re: Displaced fracture of the radius with complete 180 degrees rotation of the radial head during closed reduction. PMID- 17399873 TI - Risk of injury of the cervical spine in sled tests in female volunteers. AB - BACKGROUND: The description of cervical spine motion and the risk to sustain a cervical spine injury is mainly based on cadaver studies. As the active influence of soft tissue is neglected in cadaver studies, our understanding of the kinematic model for whiplash is limited. Therefore the objective of this study was, to describe the in vivo cervical spine motion and acceleration during rear end sled collisions to identify sequences of motion in which the risk of injury to the cervical spine is increased. METHODS: A rear-end collision with a speed change (DeltaV) of 6.3 km/h was simulated in a sled test with eight female subjects with no history of prior injury or pain of the cervical spine. A high speed camera was used to document motion data. Acceleration data were recorded using accelerometers. Acceleration input to the cervical spine was assessed by the simultaneous analysis of the head angle and angular head acceleration as well as the simultaneous analysis of the relative motion and the relative acceleration between the head and T1 to define intervals of increased risk of cervical spine injuries during rear-end collision. FINDINGS: The motion sequence is characterized by the same phases that have already been described for male volunteers. Increasing angular head acceleration can explain facet joint injuries during the extension movement (100-120 ms) and hence occur about 50 ms later than shown in cadaver models. In the late rebound the combination of maximal ventral head acceleration and head movement is underestimated and can be responsible for soft tissue injuries. INTERPRETATION: The study shows that during the extension phase and the late rebound phase, acceleration and movement pattern occur that could lead to cervical spine injuries. PMID- 17399874 TI - Bioaccumulation of trace elements in omnivorous amphibian larvae: implications for amphibian health and contaminant transport. AB - Despite the influence that amphibians have on the flow of energy and nutrients in ecological systems, the role that amphibians play in transporting contaminants through food webs has received very little attention. This study was undertaken to investigate bioaccumulation of trace elements in amphibians relative to other small aquatic organisms in a contaminated wetland. We collected bullfrog larvae (Rana catesbeiana) along with three other species of small vertebrates and four species of invertebrates from a site contaminated with a wide array of trace elements and analyzed them for trace element concentrations and stable nitrogen and carbon isotope composition. We found that amphibian larvae accumulated the highest concentrations of most trace elements, possibly due to their feeding ecology. These results suggest that omnivorous amphibian larvae can serve as a critical link for trace element trophic transfer. Their propensity to accumulate trace elements may have important implications for amphibian health in contaminated environments and should be further investigated. PMID- 17399875 TI - New methodology to investigate potential contaminant mass fluxes at the stream aquifer interface by combining integral pumping tests and streambed temperatures. AB - The spatial pattern and magnitude of mass fluxes at the stream-aquifer interface have important implications for the fate and transport of contaminants in river basins. Integral pumping tests were performed to quantify average concentrations of chlorinated benzenes in an unconfined aquifer partially penetrated by a stream. Four pumping wells were operated simultaneously for a time period of 5 days and sampled for contaminant concentrations. Streambed temperatures were mapped at multiple depths along a 60m long stream reach to identify the spatial patterns of groundwater discharge and to quantify water fluxes at the stream aquifer interface. The combined interpretation of the results showed average potential contaminant mass fluxes from the aquifer to the stream of 272microgm( 2)d(-1) MCB and 71microgm(-2)d(-1) DCB, respectively. This methodology combines a large-scale assessment of aquifer contamination with a high-resolution survey of groundwater discharge zones to estimate contaminant mass fluxes between aquifer and stream. PMID- 17399876 TI - Remediation of metal contaminated soil with mineral-amended composts. AB - This study examined the use of two composts derived from green waste and sewage sludge, amended with minerals (clinoptilolite or bentonite), for the remediation of metal-contaminated brownfield sites to transform them into greenspace. Soils contaminated with high or low levels of metals were mixed with the mineral enhanced composts at different ratios and assessed by leaching tests, biomass production and metal accumulation of ryegrass (Lolium perenne L.). The results showed that the green waste compost reduced the leaching of Cd and Zn up to 48% whereas the composted sewage sludge doubled the leachate concentration of Zn. However, the same soil amended with composted sewage sludge showed an efficient reduction in plant concentrations of Cd, Cu, Pb or Zn by up to 80%. The results suggest that metal immobilisation and bioavailability are governed by the formation of complexes between the metals and organic matter. The amendment with minerals had only limited effects. PMID- 17399877 TI - African-American men's perceptions about prostate cancer: implications for designing educational interventions. AB - This qualitative study explores African-American men's perceptions about prostate cancer (CaP) screening and assesses the acceptability of various strategies and settings for interventions to promote informed decision-making. We conducted four focus groups among healthy men (n=37) and two groups among CaP survivors (n=14) aged 35-70 in the greater Boston area, USA. Also, we conducted 14 in-depth interviews with key community informants. The audio-taped focus groups and interviews were transcribed, coded, and analyzed for emergent themes. Except for survivors, men had insufficient information about the prostate, the elevated cancer risk among African-Americans, and the controversy concerning screening. Key informants and focus group participants cited inadequate access to services, mistrust of the health system, poor relationships with medical providers, and perceived threats to male sexuality as major barriers to receiving prostate care. They recommended that interventions be embedded in community settings, address men's overall health, and be administered by culturally competent providers, and repeatedly emphasized trust building and a sustained presence in the community. Efforts to present balanced information about CaP screening may be hindered by lingering mistrust of the medical system, poor relationships between patients and providers, and enthusiastic support for screening on the part of CaP survivors. Implications for interventions are discussed. PMID- 17399878 TI - Discourses of normality and difference: responses to diagnosis and treatment of gynaecological cancer of Australian women. AB - By comparison to other cancers such as breast and lung cancer, women in Australia are relatively infrequently diagnosed with gynaecological cancers. Apart from cervical cancer, public health information on gynaecological cancer is limited, as are published stories from gynaecological cancer survivors in women's magazines. Our qualitative study investigated how women with gynaecological cancers develop an identity in relation to their illness, and examined the extent of, and reasons for, a sense of perceived difference. The study was conducted between 2001 and 2003 and included in-depth interviews with 52 women aged 27-80 years diagnosed with gynaecological cancer within the past 5 years. Our analysis illustrates how women draw on a wider cancer discourse to make sense of their own illness, which gave them a sense of commonality. However, some women, predominantly those who were diagnosed with cancer of the vulva or vagina, or who underwent particular uncommon or unfamiliar treatments such as brachytherapy, had difficulties situating their illness within the wider cancer discourse. This had implications for women when accessing social support. PMID- 17399879 TI - When time is more than money: the allocation of time between work and leisure in HIV-infected patients. AB - Time is a limited resource and individuals have to decide how many hours they should allocate to work and to leisure activities. Differences in wage rate or availability of non-labour income (financial support from families and savings) may influence how individuals allocate their time between work and leisure. An increase in wage rate may induce income effects (leisure time demanded increases) and substitution effects (leisure time demanded decreases) whereas an increase in non-labour income only induces income effects. We explored the effects of differences in wage rate and non-labour income on the allocation of time in HIV infected patients. Patients enrolled in the Swiss HIV Cohort Study (SHCS) provided information on their time allocation, i.e. number of hours worked in 1998. A multinomial logistic regression model was used to test for income and substitution effects. Our results indicate that (i) the allocation of time in HIV infected patients does not differ with level of education (i.e., wage rate), and that (ii) availability of non-labour income induces income effects, i.e. individuals demand more leisure time. PMID- 17399880 TI - On the transfer of the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) from the sow to the full-term piglet during the last third of gestation. AB - Pregnant sows were fed either a control diet (CON, n=8, 0.21 mg DON and 0.004 mg ZON/kg diet) or a diet containing 40% of a Fusarium toxin contaminated wheat (MYCO, n=7, 9.57 mg DON and 0.358 mg ZON/kg diet) from day 75 to 110 of gestation. Piglets were delivered by Caesarean section at the end. Spleen weights of piglets from the MYCO group were significantly lower. Hemoglobin concentration and hematocrit were also significantly decreased in these piglets, although this effect was more obvious in female than in male piglets. The transfer of DON and ZON was evaluated by the diet ratio (sum of concentrations of all metabolites in the physiological specimen divided by the dietary toxin concentration) and the piglet ratio (sum of concentrations of all metabolites in the physiological specimen of the piglet divided by that of the sows). The diet ratio for the liver (sows only) amounted to 0.001 (DON+de-epoxy-DON) and 0.016 (ZON and metabolites). The diet ratios of DON in bile reached up to 0.041 and 0.003 for sows and piglets, respectively, and those for ZON up to 2.896 and 0.128. The piglet ratios in bile varied up to 0.309 and 0.518 for DON and ZON, respectively, whereas nearly similar DON concentrations were found in serum of piglets and sows (median piglet ratio of 0.750). The results of the study suggest that the developing fetus is exposed to DON, ZON and their metabolites when the sows are fed a Fusarium toxin contaminated diet. PMID- 17399881 TI - The effect of nicotine in vitro on the integrity of tight junctions in Caco-2 cell monolayers. AB - Ulcerative colitis is characterised by impairment of the epithelial barrier and tight junction alterations resulting in increased intestinal permeability. UC is less common in smokers with smoking reported to decrease paracellular permeability. The aim of this study was thus to determine the effect of nicotine, the major constituent in cigarettes and its metabolites on the integrity of tight junctions in Caco-2 cell monolayers. The integrity of Caco-2 tight junctions was analysed by measuring the transepithelial electrical resistance (TER) and by tracing the flux of the fluorescent marker fluorescein, after treatment with various concentrations of nicotine or nicotine metabolites over 48 h. TER was significantly higher compared to the control for all concentrations of nicotine 0.01-10 microM at 48 h (p<0.001), and for 0.01 microM (p<0.001) and 0.1 microM and 10 microM nicotine (p < 0.01) at 12 and 24 h. The fluorescein flux results supported those of the TER assay. TER readings for all nicotine metabolites tested were also higher at 24 and 48 h only (p < or = 0.01). Western blot analysis demonstrated that nicotine up-regulated the expression of the tight junction proteins occludin and claudin-1 (p < or = 0.01). Overall, it appears that nicotine and its metabolites, at concentrations corresponding to those reported in the blood of smokers, can significantly improve tight junction integrity, and thus, decrease epithelial gut permeability. We have shown that in vitro, nicotine appears more potent than its metabolites in decreasing epithelial gut permeability. We speculate that this enhanced gut barrier may be the result of increased expression of claudin-1 and occludin proteins, which are associated with the formation of tight junctions. These findings may help explain the mechanism of action of nicotine treatment and indeed smoking in reducing epithelial gut permeability. PMID- 17399882 TI - Interactions between aflatoxin B1 and dietary iron overload in hepatic mutagenesis. AB - BACKGROUND/AIM: Dietary aflatoxin B(1) (AFB(1)) exposure and iron overload are important causes of hepatocellular carcinoma in sub-Saharan Africa. The aim of this study was to investigate if the two risk factors have an interactive effect. METHODS: Four groups of Wistar albino rats were studied for 12 months. Group 1 (control) was fed the normal chow diet; group 2 (Fe) was supplemented with 0.75% ferrocene iron; group 3 (Fe+AFB(1)) was fed 0.75% ferrocene throughout and gavaged 25 microg AFB(1) for 10 days; group 4 (AFB(1)) was gavaged 25 microg AFB(1) for 10 days. Iron profile, lipid peroxidation (LPO), 8 hydroxydeoxyguanosine (8OHdG), oxidative lipid/DNA damage immunohistochemistry, superoxide/nitrite free radicals, cytokines IL6, IL-10, transaminases (ALT/AST) and Ames mutagenesis tests were performed. RESULTS: LPO and ALT showed a significant (p<0.05)/additive effect and 8OHdG a significant (p<0.05)/multiplicative effect in the Fe+AFB(1) group. IL-6 produced a negative synergy as against an additive antagonistic effect with IL-10. Massive deposits of 4-hydroxynonenal (4-HNE) and 8OHdG were observed in liver sections of the Fe+AFB(1) group, suggestive of multiplicative synergy. Significant levels of mutagenesis (p<0.001) were observed in the Fe+AFB(1) group. This multiplicative synergy was five-fold. CONCLUSION: Dietary iron overload and AFB(1) have a multiplicative effect on mutagenesis. PMID- 17399884 TI - The missing link between action and cognition. AB - The study of the neural correlates of motor behaviour at the systems level has received increasing consideration in recent years. One emerging observation from this research is that neural regions typically associated with cognitive operations may also be recruited during the performance of motor tasks. This apparent convergence between action and cognition - domains that have most often been studied in isolation - becomes especially apparent when examining new complex motor skills such as those involving sequencing or coordination, and when taking into account external (environment-related) factors such as feedback availability and internal (performer-related) factors such as pathology. Neurally, overlap between action and cognition is prominent in frontal lobe areas linked to response selection and monitoring. Complex motor tasks are particularly suited to reveal the crucial link between action and cognition and the generic brain areas at the interface between these domains. PMID- 17399885 TI - Modeling water and sediment contamination of Lake Pontchartrain following pump out of Hurricane Katrina floodwater. AB - Levee failure and overtopping as a result of Hurricane Katrina caused major flooding of New Orleans, Louisiana. Floodwaters, which were contaminated with heavy metals, organic chemicals, and fecal coliform bacteria (FCB), were pumped into neighboring Lake Pontchartrain during dewatering. The impact of levee failure on water and benthic sediment concentrations in the lake was investigated by applying a numerical water quality model coupled to a three-dimensional, numerical hydrodynamic model. The model was used to compute water and benthic sediment concentrations throughout the lake for lead, arsenic, benzo(a)pyrene (BaP), and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), and water concentrations for FCB. Computed concentrations resulting from actual pumped discharges with levee failure and overtopping were compared to computed concentrations resulting from pumped discharges without levee failure or overtopping, and concentrations from both sets of conditions were compared to ecological water and sediment quality screening guideline values. The model indicated that incremental increases above pre-Katrina benthic sediment concentrations are about a factor of 10 greater with dewatering of the floodwaters than with dewatering of storm water without flooding. However, these increases for the metals are small relative to pre-Katrina concentrations. The results showed that the ecological screening-level sediment quality guideline values were exceeded for BaP and DDE in areas near the south shoreline of the lake as a result of floodwater pump-out, whereas, this was not the case for storm water removal without flooding. The model showed that lake water column concentrations should be about the same during both dewatering conditions regardless of whether there is flooding or not. PMID- 17399883 TI - The IRF family, revisited. AB - Since the discovery of interferon 50 years ago a great deal of progress has been made in understanding how interferons work and how and why they are induced. Key factors in interferon induction are the interferon regulatory factors (IRF). In this review of IRF we aim to show you not only the historical side of the IRF but also the integral, anti-viral and hematopoetic roles of these transcription factors, as well as the sometimes surprising and even forgotten roles that these proteins play, not only in interferon signaling but throughout the immune system and the body as a whole. Further research will no doubt expand the repertoire of these multifunctional proteins even more. PMID- 17399887 TI - Re: Stephen J. Bromage, Debbie A. Falconer, Brian A. Lieberman, et al. Sperm retrieval rates in subgroups of primary azoospermic males. Eur urol 2007;51:534 40. PMID- 17399888 TI - Re: Chao-Yu Hsu, Steven Joniau, Raymond Oyen et al. Outcome of surgery for clinical unilateral T3a prostate cancer: a single-institution experience. Eur urol 2007;51:121-9. PMID- 17399889 TI - Obesity and prostate cancer: a role for adipokines. AB - OBJECTIVES: Many studies have investigated the association between obesity and prostate cancer risk but have yielded inconsistent results. Recent evidence suggests a particular role for obesity in prostate cancer progression. Many studies have investigated the roles of adipose tissue-derived factors (adipokines) as putative molecular mediators between obesity and prostate cancer. This review provides an overview of current evidence that supports such a role for adipokines. METHODS: A comprehensive literature review was carried out using PubMed to search for articles relating to prostate cancer and the following adipokines: leptin, interleukin 6, vascular endothelial growth factor (VEGF), and adiponectin. RESULTS: Prostate cancer cells are exposed to adipokines either via the circulation or through locally produced adipokines following invasion of the retropubic fat pad. Circulating levels of most adipokines are positively correlated with obesity; adiponectin is inversely correlated with obesity. High circulating levels of leptin, interleukin 6, and VEGF are associated with increased prostate cancer risk and increased aggressiveness. Adiponectin levels are lower in patients with prostate cancer and are inversely associated with grade of disease. Adipokines exert a variety of biologic effects on prostate cancer cells, modulating cellular differentiation, apoptosis, proliferation, and angiogenesis. CONCLUSIONS: Evidence suggests a role for obesity and adipokines in promoting the progression of established prostate cancer. Adipokines may contribute to the molecular basis for the association between obesity and prostate cancer, but the complex pathophysiology of both these disease states requires further studies. PMID- 17399890 TI - New procedures are bound to modify our attitude in facing urologic cancers in the near future: are we ready for a cost-effectiveness analysis? PMID- 17399891 TI - Four cardiac hormones eliminate 4-fold more human glioblastoma cells than the green mamba snake peptide. AB - Within 24h four cardiac hormones, i.e., vessel dilator, kaliuretic peptide, atrial natriuretic peptide, and long acting natriuretic peptide decrease the number of human glioblastoma cells 75%, 68%, 67%, and 65% while Dendroaspis (green mamba) peptide caused a 17% decrease when each were utilized at 100 microM. The four cardiac hormones decreased DNA synthesis 65-87% and increased cyclic GMP 1.3- to 3.8-fold in the glioblastoma cells. Natriuretic peptide receptors (NPR)-A and -C were present. CONCLUSION: four cardiac hormones eliminate up to 75% of glioblastoma cells via cyclic GMP-mediated up to 87% decrease in DNA synthesis. PMID- 17399892 TI - Diet-induced obesity and mammary tumor development in relation to estrogen receptor status. AB - Leptin enhances proliferation of estrogen receptor (ER)-positive breast cancer cells in vitro. Here, we compared mammary tumor (MT) formation from ER-positive (MCF-7) and ER-negative (MDA-MB-231) breast cancer cells in athymic mice fed a High-Fat diet to elevate serum leptin. Neither body weight, diet or serum leptin levels impacted MT latency, burden or tumor grade. However, protein expression in mammary fat pads exhibited elevated PCNA and Cyclin D1 while in MTs, Ob-Rb, IGF IR, Bcl-2, and Bax were lower in Low-Fat versus High-Fat mice. In conclusion, diet rather than serum leptin impacted breast cancer cell tumor metabolism. PMID- 17399893 TI - Adsorption of arsenate on untreated dolomite powder. AB - Raw dolomite powder was evaluated for its efficiency in adsorbing As(V) from water. An experimental setup comprised of a fluidized dolomite powder bed was used to assess the impact of various test variables on the efficiency of removal of As(V). Test influents including distilled water (DW), synthetic groundwater (SGW) and filtered sewage effluent (FSE) were employed to assess the effect of influent parameters on the adsorption process and the quality of the effluent generated. Dolomite exhibited good As(V) removal levels for distilled water (>92%) and synthetic ground water (>84%) influents at all initial As(V) concentrations tested (0.055-0.600 ppm). Breakthrough of dolomite bed occurred after 45 bed volumes for DW and 20 bed volumes for SGW influents with complete breakthrough taking place at more than 300 bed volumes. As(V) removal from FSE influents was relatively unsuccessful as compared to the DW and SGW influents. Partial removal in the order of 32% from filtered sewage effluent at initial concentration of 0.6 mg/L started at 75 bed volumes and gradually stopped at 165 bed volumes. Varying degrees of As(V) adsorption capacities were observed by the different test influents employed, which indicate that the adsorption of As(V) is adversely affected by competing species, mainly sulfates and phosphates present in the influent. The adsorptive behavior of dolomite was described by fitting data generated from the study into the Langmuir and Freundlich isotherm models. Both models described well the adsorption of dolomite. The average isotherm adsorptive capacity was determined at 5.02 mug/g. Regeneration of the dolomite bed can be achieved with the use of caustic soda solution at a pH of 10.5. PMID- 17399894 TI - Boundary conditions for developing a safety concept for an exothermal reaction. AB - Kinetic calculations for an example exothermal chemical process, the production of TCB, are carried out. They address both parameter uncertainties and random failures of the cooling system. In this way, they enable one to establish comprehensive boundary conditions for a safety system in terms of unavailability, the quantities of the undesired by-product (TCDD) produced and the times available before a required intervention, if a pre-determined quantity of TCDD is tolerated. It is shown that accounting for stochastic effects and uncertainties derived from insufficient knowledge provides a broader and more realistic knowledge base for devising a viable safety concept. PMID- 17399895 TI - Distribution of chloride ion in MSWI bottom ash and de-chlorination performance. AB - When recycling bottom ash from municipal solid waste incinerators (MSWIs), salts and heavy metals contents must be considered; in particular, chloride ions must be addressed because they cause serious corrosion in metals. Therefore, only limited amounts of bottom ash can be utilized as a substitution for material or the bottom ash must be treated at high temperatures prior to use. These factors markedly decrease the applications of bottom ash. In addition to the distribution characteristics of chloride ions, this study also investigates the characteristics change before and after de-chlorination using a counter-flow pipe column and three different flow fluxes for different refuse incinerators as the experiment variables. Thus, this study attempts to determine the appropriate conditions for de-chlorination and an appropriate policy for use of bottom ash as concrete aggregate. The experimental results show that a negative correlation exists between the natural logarithm of the chloride ion concentration and particle size in bottom ash. Characteristics of de-chlorinated bottom ash, such as pH value, mud content, loss on ignition, chloride ion concentration, turbidity, and species intensity, all decrease, meaning that de-chlorination decreased chloride ion content and generates a cleaning effect. The per-unit-time efficiency of de-chlorination is highest in the high flux flow. When flow flux is 80 mL/min, the de-chlorination efficiency is >0.3%/h. However, the shortest time required for bottom ash de-chlorination does not reduce in proportion to the legally prescribed concentration of chloride ion. PMID- 17399896 TI - Biochanin A protects dopaminergic neurons against lipopolysaccharide-induced damage through inhibition of microglia activation and proinflammatory factors generation. AB - Activation of microglia and consequent release of proinflammatory factors, are believed to contribute to neurodegeneration in Parkinson's disease (PD). Hence, identification of compounds that prevent microglial activation is highly desirable in the search for therapeutic agents for inflammation-mediated neurodegenerative diseases. In this study, we reported that biochanin A, one of the predominant isoflavones in Trifolium pratense, attenuated lipopolysaccharide (LPS)-induced decrease in dopamine uptake and the number of dopaminergic neurons in a dose-dependent manner in rat mesencephalic neuron-glia cultures. Moreover, biochanin A also significantly inhibited LPS-induced activation of microglia and production of tumor necrosis factor-alpha, nitric oxide and superoxide in mesencephalic neuron-glia cultures and microglia-enriched cultures. This study suggested for the first time that biochanin A protected dopaminergic neurons against LPS-induced damage through inhibition of microglia activation and proinflammatory factors generation. PMID- 17399897 TI - p53 is upregulated in Alzheimer's disease and induces tau phosphorylation in HEK293a cells. AB - p53 and tau are both associated with neurodegenerative disorders. Here, we show by Western blotting that p53 is upregulated approximately 2-fold in the superior temporal gyrus of Alzheimer's patients compared to healthy elderly control subjects. Moreover, p53 was found to induce phosphorylation of human 2N4R tau at the tau-1/AT8 epitope in HEK293a cells. Confocal microscopy revealed that tau and p53 were spatially separated intracellularly. Tau was found in the cytoskeletal compartment, whilst p53 was located in the nucleus, indicating that the effects of p53 on tau phosphorylation are indirect. Collectively, these findings have ramifications for neuronal death associated with Alzheimer's disease and other tauopathies. PMID- 17399898 TI - Time course of attentional modulations on automatic emotional processing. AB - In a previous study using event-related potentials (ERPs) [S. Doallo, S. Rodriguez Holguin, F. Cadaveira, Attentional load affects automatic emotional processing: evidence from event-related potentials, Neuroreport 17 (2006) 1797 1801], we reported that differential responses to unattended peripheral affective pictures, as reflected by N1-P2 modulations at posterior regions, are modulated by attentional load at fixation. Here, new analyses of these data were performed to evaluate whether a sustained, broadly distributed, negative shift in the unattended pictures ERP waveforms, which displayed larger amplitudes for emotional stimuli, reflects an additional differential response to the emotional content. Under low-load conditions, unpleasant (versus neutral) pictures elicited greater negativities in the 80-140 ms latency range over frontocentral sites and more centroparietally distributed from 200 to 280 ms. These findings provide further evidence of the time course of emotional processing at unattended locations and its modulation by attentional load. PMID- 17399899 TI - Increase of ceramide monohexoside and dipalmitoyl glycerophospholipids in the brain of Zellweger syndrome. AB - The lipid composition and molecular species of phospholipids were examined in the brain of a patient with Zellweger syndrome (ZS), and were compared with those of control infants. In the cerebral gray matter of the ZS patient, the amounts of ceramide monohexoside and cholesterol ester were larger than those of controls. By contrast, the amount of ceramide monohexoside in the white matter was smaller in the ZS patient than that in the age-matched control. Although the amount of phosphatidylcholine (PC) plus phosphatidylserine (PS) was the same, dipalmitoyl PC and PS were increased in both the gray and white matter of the ZS cerebrum. These alterations in the molecular species of brain lipids may play crucial roles in the pathogenesis of ZS. PMID- 17399900 TI - Functional dysconnectivity of the dorsolateral prefrontal cortex in first-episode schizophrenia using resting-state fMRI. AB - The known regional abnormality of the dorsolateral prefrontal cortex (DLPFC) and its role in various neural circuits in schizophrenia has given prominence to its importance in studies on the dysconnection associated with schizophrenia. Abnormal functional connectivities of the DLPFC have been found during various goal-directed tasks; however, the occurrence of the abnormality during rest in patients with schizophrenia has rarely been reported. In the present study, we selected bilateral Brodmann's area 46 as region of interest and analyzed the differences in the DLPFC functional connectivity pattern between 17 patients with first-episode schizophrenia (FES) and 17 matched controls using resting-state fMRI. We found that the bilateral DLPFC showed reduced functional connectivities to the parietal lobe, posterior cingulate cortex, thalamus and striatum in FES patients. We also found enhanced functional connectivity between the left DLPFC and the left mid-posterior temporal lobe and the paralimbic regions in FES patients. Our results suggest that functional dysconnectivity associated with the DLPFC exists in schizophrenia during rest. This may be partially related to disturbance in the intrinsic brain activity. PMID- 17399901 TI - Some aspects of the silicon behaviour under femtosecond pulsed laser field evaporation. AB - Three dimension atom probe analysis of semiconductor materials requires the ability to bring high electric field at the specimen apex to remove atoms. It is shown that, if voltage pulses are used to evaporate doped silicon, the resistivity of the material has to be lower than about 10(2) Omega cm. To overcome this problem, voltage pulses have been replaced by femtosecond laser pulses. The laser pulses give rise to field evaporation by two processes. Both thermal and optical field evaporation have been observed. Thermal evaporation takes place at high laser intensities and with short wavelengths while the evaporation is assisted by the rectification of the optical field for lower intensities and in the infrared domain. Using the optical field evaporation, reproducible and good analyses in term of spatial and mass resolutions could be conducted. PMID- 17399902 TI - The influence of relativistic energy losses on bandgap determination using valence EELS. AB - Since monochromated transmission electron microscopes have become available, the determination of bandgaps and optical properties using electron energy loss spectrometry (EELS) has again attracted interest. The underlying idea is very simple: below the bandgap energy no transitions can contribute to the valence EELS signal. However, the bandgap cannot be directly read out from the recorded data. Therefore the optical properties cannot be determined correctly from the low loss using the Kramers-Kronig relations. We will discuss under which conditions relativistic effects may be suppressed. It is demonstrated that scanning TEM (STEM) geometry is not applicable for most bandgap measurements. PMID- 17399904 TI - Ultrastructural changes in Tritrichomonas foetus after treatments with AlPcS4 and photodynamic therapy. AB - The Tritrichomonas foetus is an amitochondrial parasitic protist which causes bovine trichomoniasis, a major sexually transmitted disease in cattle. No effective drugs for this disease have been approved to this date. Photodynamic therapy (PDT) is an experimental treatment that shows great potential for treating bacteria, fungi, yeasts, and viruses. However, the cytotoxic effect of PDT on protozoan has been poorly studied. In this study, PDT with aluminum phthalocyanine tetrasulfonated (AlPcS4) photosensitizer was efficient in killing T. foetus. The mode of cell death in T. foetus after PDT was investigated by transmission electron microscopy. Morphological changes, such as membrane projections, nucleus fragmentation with peripheral masses of heterochromatin, endoplasmic reticulum proliferation, intense cytoplasmic vacuolization, fragmented axostyle-pelta complex, and internalized flagella could be observed. This is the first report to demonstrate cell death in T. foetus after PDT, and thus will open up new lines of investigation to develop new treatments for bovine trichomoniasis. PMID- 17399905 TI - Role of plasma and liver cholesterol- and lipoprotein-metabolism determinants in LpX formation in the mouse. AB - Cholestasis is characterized by hypercholesterolemia and the appearance of an abnormal lipoprotein, lipoprotein X (LpX), in plasma. The mechanisms responsible for this cholestatic plasma lipid phenotype are not fully understood. We used ATP binding cassette A1 (ABCA1)-/- and scavenger receptor class B type I (SR-BI)-/- mice to test the hypothesis that hepatic sinusoidal cholesterol transporters contribute to LpX formation and hypercholesterolemia during cholestasis. Bile duct ligation (BDL) of both ABCA1-/- and SR-BI-/- mice, as well as their respective controls, induced a dramatic increase in plasma cholesterol and phospholipid concentrations. Plasma fractionation revealed the presence of LpX in plasma of cholestatic mice, irrespective of their genetic background. We observed that the presence of HDL before cholestasis, a decrease in the activity of LCAT, and an increase in VLDL synthesis were not required for hypercholesterolemia and lipoprotein modifications induced by obstructive cholestasis in mice. In addition, murine cholestasis resulted in increased hepatic cholesterol synthesis that may contribute to the higher plasma free cholesterol levels found during the early hours after BDL. Together these findings indicate that hypercholesterolemia and LpX formation associated with obstructive cholestasis are correlated with an increase in hepatic cholesterol synthesis and are independent of plasma HDL levels, LCAT activity, VLDL synthesis, and ABCA1 and SR-BI expression. PMID- 17399906 TI - A putative helical cytokine functioning in innate immune signalling in Drosophila melanogaster. AB - In invertebrates and vertebrates, innate immunity is considered the first line of defense mechanism against non-self material. In vertebrates, cytokines play a critical role in innate immune signalling. To date, however, the existence of genes encoding for invertebrate helical cytokines has been anticipated, but never demonstrated. Here, we report the first structural and functional evidence of a gene encoding for a putative helical cytokine in Drosophila melanogaster. Functional experiments demonstrate that its expression, as well as that of the antimicrobial factors defensin and cecropin A1, is significantly increased after immune stimulation. These observations suggest the involvement of helical cytokines in the innate immune response of invertebrates. PMID- 17399907 TI - Increases in impulsivity following smoking abstinence are related to baseline nicotine intake and boredom susceptibility. AB - Trait impulsivity and response inhibition have been shown to be related to smoking behavior. One measure of response inhibition - antisaccade performance, or the ability to inhibit looking at a novel stimulus - has been shown to be worsened by smoking abstinence, improved by nicotine administration and predictive of smoking cessation outcomes. However, relations between antisaccade performance and measures of trait impulsivity have not been extensively evaluated in smokers. In the present study, twelve dependent smokers (n=12) completed an eye tracking task following smoking as usual and overnight abstinence; and they completed baseline measures of trait impulsivity, smoking history and provided biological samples. As expected, overnight abstinence significantly increased antisaccade errors (p<0.002) while having no effect on prosaccade performance. Abstinence-induced increases in antisaccade errors were positively correlated with baseline plasma cotinine and Sensation Seeking Scale Boredom Susceptibility, and negatively correlated with IQ. These results suggest that smoking abstinence significantly increases errors of response inhibition and that the magnitude of this increase is related to trait impulsivity and nicotine intake variables. PMID- 17399908 TI - Multi-episode survival analysis: an application modelling readmission rates of heroin dependents at an inpatient detoxification unit. AB - The purpose of this study is to describe the characteristics of a statistical technique appropriate for analysing multi-episode data (multi-episode survival analysis), and to show its application in modelling the flow of readmissions at an inpatient detoxification unit. Data are from 784 opioid-dependent patients admitted at an inpatient detoxification unit, who totalled 1,255 admission episodes. Information stored prospectively at the unit database was reviewed for the following variables at the time of each patient discharge: episode serial number, sex, route of heroin administration, reason for discharge, time of discharge, and transition time (re-entry into the inpatient detoxification unit). Cox's semi-parametric regression model seems the most appropriate for describing the series of episodes. Amongst the parametric models, most noteworthy was the superior fit of the Gompertz-Makeham model, suggesting that the transition rate decreases monotonically with time. The influence of the variables assessed differed based on the serial number of the episode. The results suggest that multi-episode survival analysis is a statistical method that can fully address the long-term perspective on treatment utilization. PMID- 17399909 TI - Nickel dust halo surrounding the guitarists: a potential trigger for the guitarists' respiratory problems. PMID- 17399910 TI - A "pancreatic tooth" design best accommodates the limitations of current artificial pancreas technology. AB - Inadequately responsive glycaemic control is an important factor in the causation of diabetic end-organ damage. Artificial or hybrid bioartificial pancreases can provide responsive glycaemic control that can reduce the enormous personal suffering and socio-economic costs of diabetes. However, they share the shortcomings of limited operational life, due to depletion of stores or failure of component parts. A pancreatic tooth design provides accessibility for the purposes of replenishment or replacement. In addition, the mouth also provides a sheltered location, is more resistant to diabetic changes and less prone to thermoregulatory changes than subcutaneous tissues, and is adapted to cope with the occasional pathogen load. The device would consist of two parts: a permanent implant with an angiogenic capillary plexus that is the blood contacting interface and a crown containing the artificial or bio-artificial pancreatic systems: the accessibility of which confers ease of replenishment and replacement, among other advantages. PMID- 17399911 TI - The potential impact of sickness-motivated behavior on the expression of neuropsychiatric disturbances in systemic lupus erythematosus. AB - Activation of the peripheral immune system is often accompanied by changes in cognition, ingestive behavior, sleep pattern, and sexual drive; collectively referred to as sickness behavior. Mounting evidence suggests that sickness behavior may be a purposeful attempt on the part of an organism to conserve energy and thereby facilitate recuperation. Illnesses characterized by chronic, uncontrolled immune reactivity such as systemic lupus erythematosus are also frequently associated with impaired emotionality and cognition; which, unlike sickness behavior, are conventionally thought to emanate from fixed structural lesions of the brain. Clinical observations, however, indicate that the neuropsychiatric disturbances in lupus may wax and wane in intensity and suggest the hypothesis that sickness-motivated behavior may significantly influence the neuropsychiatric manifestations of systemic lupus erythematosus and, perhaps, those of other autoimmune diseases associated with neuroinflammation. The hypothesis that patients with systemic lupus erythematosus undergo a reorganization of their motivational priorities, which influences cognitive performance and emotional output, may be examined using validated behavior paradigms in autoimmune MRL-MpJ-Tnfrsf6(lpr) (MRL-lpr/lpr) mice that spontaneously develop a lupus-like illness accompanied by disturbances in cognition and emotionality. Confirming that sickness-motivated behavior contributes to the aberrations in cognition and emotionality exhibited by an experimental model of systemic lupus erythematosus might have important therapeutic and prognostic implications by invoking the possibility that similar motivational effects may be influencing cognitive and/or emotional output in patients with neuropsychiatric lupus. PMID- 17399912 TI - Increased cell hydration promotes both tumor growth and metastasis: a biochemical mechanism consistent with genetic signatures. AB - It was postulated previously that a progressive increase in cell hydration, induced by successive genetic or epigenetic changes, is the basic mechanism of multistep carcinogenesis, and also that the degree of malignancy increases with the degree of cell hydration. These hypotheses implied that increased cell hydration is a common factor promoting both tumor growth and metastasis, and that metastatic potential increases with the degree of cell hydration. This paper discusses these implications in relation to current concepts of genetic mechanisms determining the acquisition of metastatic potential. It was also postulated previously that the enhancement of metabolic activity by increased cell hydration will increase the ability of tumor cells to compete for nutrients with their normal counterparts. This effect may favor the preferential selection of cells whose genotypes confer the greatest increase in cell hydration and which, on the present hypothesis, would be those with the greatest capacity for metastasis. An important feature of this "common factor" hypothesis is that it suggests a biochemical explanation for DNA-microarray data showing a similarity between the gene expression patterns associated with both tumor growth and metastasis, while the postulated role of genes causing increased cell hydration might explain the apparent acquisition of metastatic potential at an early stage of tumorigenesis. Previous investigations were consistent with the hypothesis that various factors promoting carcinogenesis may do so by increasing cell hydration. A survey of the literature showed that all of these factors also promote cell motility, migration or metastasis, and provided evidence that these effects could be attributed to the associated increase in cell hydration. Methods are suggested for testing the hypothesis, and the paper concludes by emphasizing the need for more research on the biochemistry of cancer, and on the role of water as a biochemical factor of particular importance, not only in carcinogenesis, but in many other aspects of cell biology. PMID- 17399913 TI - [Primary ovarian leiomyosarcoma in perimenarche]. AB - Primary ovarian leiomyosarcoma is an extremely rare tumor, which is thought to arise predominantly in postmenopausal women. We report the second case in a perimenarchal adolescent. Diagnosis was grounded on light microscopic and histochemical findings. In our case, the treatment was exclusively surgical. As there was no tumoral residue, and according to the low-grade tumor, no adjuvant treatment was performed. PMID- 17399914 TI - [Reproductive outcome after laparoscopic treatment of endometriosis in an infertile population]. AB - OBJECTIVES: To evaluate fertility outcome after laparoscopic management of endometriosis in an infertile population. MATERIALS AND METHODS: A retrospective analysis of 64 patients presenting more than one year infertility and a pregnancy wish associated with minimal to severe endometriotic lesions (stage I to IV according to the revised American Fertility Society (rAFS) classification), treated using laparoscopic surgery in order to remove the entire lesions. We excluded women under 20 years and over 40, as well as those with other infertility factors (tubal non endometriosis-related, hormonal or sperm). Fertility of the remaining 34 patients was studied in relation to endometriosis stage and to pregnancy's mode (spontaneous or induced). RESULTS: Pregnant women percentage was 65% (22 patients) within a 8.5 months (quartiles: 3; 15.5) [range: 1; 52] post-surgical time, and 86.5% pregnancies issued with a delivery. The rate of pregnant women depended on stage of endometriosis (89% for stages I-II, and 56% for stages III-IV). Sixty percent pregnancies were spontaneous within a 5 months (3; 9) [1; 52] post-surgical time to pregnancy average. When pregnancies were obtained with assisted reproductive techniques, the median post-surgical time to pregnancy was 12 months (9; 22) [2; 31]. Among women with stages I-II endometriosis, the median post-surgical time to pregnancy was 2 months when spontaneous and 20.5 months when induced (P=0.007). In case of stages III-IV endometriosis, pregnancy's delay was 8 and 12 months respectively (P=0.79). Among the 21% women who had had an induced pregnancy failure before surgery, 71% became pregnant and 80% spontaneously. Eighteen patients (53%) had an ovarian endometrioma and 50% of them became pregnant. Among the 4 patients who had colorectal endometriosis requiring colorectal resection, 1 pregnancy was obtained. CONCLUSIONS: These findings suggest that in a context of more than one year infertility only related to endometriosis, it is reasonable to offer these patients a complete operative laparoscopic treatment of their lesions, which enables 65% of them to be pregnant within a 8.5 months post-surgical median time to pregnancy and spontaneously in 60%. In case of stages I-II endometriosis we suggest a spontaneous pregnancy try during 8 to 12 months before starting induced pregnancy therapeutics instead of stages III-IV endometriosis where induced methods should be used after only 6 or 8 months. PMID- 17399915 TI - Undermatching is an emergent property of selection by consequences. AB - A computational theory of selection by consequences [McDowell, J.J, 2004. A computational model of selection by consequences. J. Exp. Anal. Behav. 81, 297 317] was tested by studying the responding of virtual organisms that were animated by the theory on random interval schedules of reinforcement. The theory generated responding by applying principles of selection, reproduction, and mutation to a population of potential behaviors that evolved in response to the selection pressure exerted by reinforcement. The organisms' equilibrium response rates were well described by the modern version of the Herrnstein hyperbola, which includes an exponent on reinforcement rate. Under strong selection pressure this exponent decreased with increasing mutation rate from a value near 1.0 at 1% mutation to an asymptotic value of 0.83 at mutation rates of 10% and greater. This asymptotic value is consistent with values obtained by fitting the equation to data from live organisms responding on single schedules, and with the value of about 0.80 that is expected on the basis of extensive research with live organisms responding on concurrent schedules. These results show that the computational theory is consistent with the modern theory of matching [McDowell, J.J, 2005. On the classic and modern theories of matching. J. Exp. Anal. Behav. 84, 111-127], and that it is a viable candidate for a mathematical dynamics of behavior. PMID- 17399916 TI - Clever Hans is still whinnying with us. PMID- 17399917 TI - Screening of pigs resistant to F4 enterotoxigenic Escherichia coli (ETEC) infection. AB - The present study analysed quantitatively the mucin 4 polymorphism for determining the F4ac/ab receptor status of a total of 63 pigs by comparing it with the in vitro villous adhesion assay. The probability of a susceptible genotype for the mucin 4 increases significantly with increasing F4ab or F4ac ETEC adhesion per 250 microm villi (P=0.029 for F4ab, P=0.030 for F4ac), with the odds ratio for each unit increase of F4ab or F4ac equal to, respectively, 1.036 (95% CI [1.004-1.069]) and 1.018 (95% CI [1.002-1.034]). In the phenotypic in vitro villous adhesion test, a cut-off value of 5 bacteria was chosen as a criteria for the distinction between an F4R positive and F4R negative pig. The sensitivity and specificity for the in vitro villous adhesion test, with the genotyping test for mucin 4 as golden standard, is 100% and 24%, respectively, for F4ab as well as F4ac. Absence of adhesion of F4ac and F4ab ETEC to the villous brush borders was not associated with genotypic resistance suggesting that there is at least one other receptor for F4ab/ac Escherichia coli. As a consequence, not only mucin 4 gene polymorphism but also expression of these other receptor(s) has to be included in a screening assay for F4ac/ab receptor negative pigs. PMID- 17399918 TI - Cigarette smoke condensate increases cathepsin-mediated invasiveness of oral carcinoma cells. AB - Cigarette smoke, which contains several carcinogens known to initiate and promote tumorigenesis and metastasis, is the major cause of oral cancer. Lysosomal cathepsin proteases play important roles in tumor progression, invasion and metastasis. In the present work we investigated the effects of cigarette smoke condensate (CSC) on cathepsin (B, D and L) expression and protease-mediated invasiveness in human oral squamous cell carcinoma (OSCC) cells. Our results show that treatment of OSCC cells (686Tu and 101A) with CSC activated cathepsins B, D and L in a dose-dependent manner. Both expression and activity of these cathepsins were up-regulated in CSC-exposed versus non-exposed cells. Although cathepsin L had the lowest basal level, it had the highest induction in exposed cells compared to cathepsins B and D. Suppression of CSC-induced cathepsin B and L activities by specific chemical inhibitors decreased the invasion process, suggesting that these proteases are involved in the invasion process. Overall, our results indicate that CSC activates cathepsin B and L proteolytic activity and enhances invasiveness in OSCC cells, a response that may play a role in CSC mediated tumor progression and metastasis dissemination. PMID- 17399919 TI - Reproductive tissue regression: involvement of caspases, inducible nitric oxide synthase and nitric oxide during moulting in White Leghorn hens. AB - Moulting is a natural physiological process where the reproductive system of birds undergoes complete remodeling in preparation for the next laying cycle. In domestic chickens, moulting is artificially induced by feed withdrawal to recycle the old laying flock for best profit margins. This has received severe criticism from animal welfare organizations, forcing several countries to stop this practice. Several alternative methods to feed withdrawal methods were developed but were found to produce inconsistent results. Understanding the actual mechanism of moulting would help in designing a new animal welfare friendly method. The present investigation attempted to study the molecular mechanism of moulting in White Leghorn hens. Eighty-four layers (75 weeks) were divided into two groups. The birds in the first group were subjected to moulting by feed withdrawal (FW) while the other group received high dietary Zn (ZnF) treatment for 10 days. Six birds from each group were sacrificed on 0, 1-4, 6 and 10 days of moulting and mRNA expression of caspases-1, -2 and iNOS, along with the apoptotic ladder pattern and nitric oxide (NO) in the ovary and oviduct, was investigated. The mRNA expression of iNOS was upregulated with a corresponding increase in NO levels. Caspases-1 and -2 were differentially upregulated in the ovary and oviduct of moulted birds. A constant decline in serum estradiol and progesterone levels was also observed. It can be concluded that the pattern of reproductive regression during moulting by the two methods is different, as the expression of genes studied in the present investigation is different. PMID- 17399920 TI - Development of bovine-ovine interspecies cloned embryos and mitochondria segregation in blastomeres during preimplantation. AB - The objective of the study was to investigate interspecies somatic cell nuclear transfer (iSCNT) embryonic potential and mitochondrial DNA (mtDNA) segregation during preimplantation development. We generated bovine-ovine reconstructed embryos via iSCNT using bovine oocytes as recipient cytoplasm and ovine fetal fibroblast as donor cells. Chromosome composition, the total cell number of blastocyst and embryonic morphology were analyzed. In addition, mtDNA copy numbers both from donor cell and recipient cytoplasm were assessed by real-time PCR in individual blastocysts and blastomeres from 1- to 16-cell stage embryos. The results indicated the following: (1) cell nuclei of ovine fetal fibroblasts can dedifferentiate in enucleated bovine ooplasm, and the reconstructed embryos can develop to blastocysts. (2) 66% of iSCNT embryos had the same number of chromosome as that of donor cell, and the total cell number of iSCNT blastocysts was comparable to that of sheep parthenogenetic blastocysts. (3) RT-PCR analysis in individual blastomeres revealed that the ratio of donor cell mtDNA: recipient cytoplasm mtDNA remained constant (1%) from the one- to eight-cell stage. However, the ratio decreased from 0.6% at the 16-cell stage to 0.1% at the blastocyst stage. (4) Both donor cell- and recipient cytoplasm-derived mitochondria distributed unequally in blastomeres with progression of cell mitotic division. Considerable unequal mitochondrial segregation occurred between blastomeres from the same iSCNT embryos. PMID- 17399921 TI - Intraglandular injection of antibiotics for the treatment of vesicular adenitis in bulls. AB - Two experiments were designed to determine the efficacy of intraglandular antibiotic treatment in beef bulls. Experiment 1 was designed to evaluate the glandular tissue reaction to intraglandular antibiotic treatment. Experiment 2 was conducted to determine the efficacy intraglandular injection of antibiotics for the treatment of naturally occurring cases of vesicular adenitis. Healthy beef bulls (n=15), 2 and 3 years of age, were randomly allocated to three equal treatment groups to receive 10% of the daily recommended parenteral dose of penicillin, ceftiofur, or oxytetracycline in a volume of 6 mL injected directly into one of the vesicular glands. Ultrasonography was performed before, immediately after, and at 24, 48 and 168 h after intraglandular injection. The size and hardness of vesicular glands injected with oxytetracycline was greater (P<0.01) than those injected with ceftiofur. Ultrasonographic pixel intensity increased (P<0.01) after treatment with antibiotics, especially after treatment with oxytetracycline or penicillin. In Experiment 2, yearling beef bulls with clinical vesicular adenitis (n=14) were referred to the Western College of Veterinary Medicine for treatment. Eight bulls had unilateral and six had bilateral vesicular adenitis. The most common isolate was Arcanobacterium pyogenes. Corynebacterium pseudotuberculosis was isolated from one bull. Bulls were subjected to rectal palpation and ultrasonography of the vesicular glands, semen collection by electroejaculation, and intraglandular treatment with ceftiofur (n=13) and if necessary, a second intraglandular treatment of penicillin (n=6). One bull was treated only with an initial intraglandular injection of penicillin. Bulls were evaluated once a week over 6 weeks by palpation of the glands, and evaluation of semen. All bulls recovered from vesicular adenitis after 3-6 weeks. There was a difference in the amount of pus (P=0.042), leukocytes (P<0.001) and blood (P=0.003) present in ejaculates from before treatment to 3 weeks after treatment. Pixel intensities in ultrasonographic images of healthy or affected vesicular glands, whether treated or untreated, did not change over time. Intraglandular injection of ceftiofur in yearling bulls via the ischiorectal fossa was effective for treating vesicular adenitis. PMID- 17399922 TI - Molecular characterization and expression profiles of cyclin B1, B2 and Cdc2 kinase during oogenesis and spermatogenesis in rainbow trout (Oncorhynchus mykiss). AB - The meiotic maturation of oocyte and spermatocyte in animals is controlled by the maturation promotion factor (MPF), a complex of Cdc2 and cyclin B proteins. To better understand the mechanism of oocyte and spermatocyte maturation in fish, the expression of cyclin B1 (CB1), B2 (CB2) and Cdc2 kinase during oogenesis and spermatogenesis in rainbow trout were examined at both the mRNA and protein levels. Quantitative real-time PCR analysis showed that the amount of CB1 and CB2 mRNA was greater at previtellogenesis and late vitellogenesis stages, but less at early vitellogenesis stage and during early embryogenesis. Cdc2 mRNA was continuously present throughout the processes of oogenesis and early embryogenesis except for a decline at early vitellogenesis. In situ hybridization analysis indicated that CB1, CB2 and Cdc2 transcripts were present in oocytes of different developmental stages as well as in all spermatogenic cells except for spermatogonia. Immunohistochemical analysis revealed that CB1 protein was absent in vitellogenic oocytes, but present in young previtellogenic and mature oocytes. In contrast, CB2 and Cdc2 proteins were present at all stages oocyte development. Similarly, CB2 and Cdc2 proteins were present throughout spermatogenesis, whereas CB1 protein was only detected in spermatogonia and spermatocytes, but not in spermatids. Thus, it appears that CB1, CB2 and Cdc2 transcripts have similar expression patterns during oogenesis and spermatogenesis, but CB1 protein varies in amount during these processes. These data suggest that CB1 may have a leading role in the regulation of meiotic maturation of oocytes and spermotocytes. PMID- 17399923 TI - The problems with some epidemiological studies. AB - Epidemiological studies provide a valuable tool for the investigation of the causes of disease. However, such studies alone are rarely able to prove cause. No study is perfect and the evaluation of results must take account of the design and execution of the study together with the analytic methods used. Before imputing cause it is important to consider the findings against the criteria set out by Bradford Hill. Some of the problems with studies are discussed including the specification of the data collected, accuracy of information, end point definition, study size and the way results are presented. The issues are illustrated by reference to published papers. It is concluded that unless studies are evaluated and interpreted with care they may result in more harm than good. PMID- 17399924 TI - Identification of subdivisions in the medial geniculate body of the guinea pig. AB - The accurate and reliable identification of subdivisions within the auditory thalamus is important for future studies of this nucleus. However, in the guinea pig, there has been no agreement on the number or nomenclature of subdivisions within the main nucleus of the auditory thalamus, the medial geniculate body (MGB). Thus, we assessed three staining methods in the guinea pig MGB and concluded that cytochrome oxidase (CYO) histochemistry provides a clear and reliable method for defining MGB subdivisions. By combining CYO with acetylcholinesterase staining and extensive physiological mapping we defined five separate divisions, all of which respond to auditory stimuli. Coronal sections stained for CYO revealed a moderate to darkly-stained oval core. This area (the ventral MGB) contained a high proportion (61%) of V-shaped tuning curves and a tonotopic organisation of characteristic frequencies. It was surrounded by four smaller areas that contained darkly stained somata but had a paler neuropil. These areas, the dorsolateral and suprageniculate (which together form the dorsal MGB), the medial MGB and the shell MGB, did not have any discernable tonotopic frequency gradient and contained a smaller proportion of V-shaped tuning curves. This suggests that CYO permits the identification of core and belt areas within the guinea pig MGB. PMID- 17399925 TI - Evaluation of antinociceptive activity of Casearia sylvestris and possible mechanism of action. AB - The antinociceptive properties of Casearia sylvestris Sw. (Flacourtiaceae) were investigated in various models of pain-related behavior in mice. The hydroalcoholic crude extract of the plant (30-300mg/kg, per os) clearly inhibited nocifensive responses induced by ovalbumin (hindpaw licking) or acetic acid (writhes) in graded fashion. At 300mg/kg, the extract reduced nocifensive behaviors (from 71.1+/-13.3 to 14.8+/-9.3s; from 31.3+/-4.5 to 3.3+/-1.2 writhes, respectively) to similar extents as indomethacin (5mg/kg; 5.7+/-1.1s and 3.3+/ 1.2 writhes, respectively). Significant antinociceptive effects in the hot plate test were only detected following administration of the highest extract dose, but this analgesic action appeared to be specific as the extract failed to change motor and exploratory activities. The antinociceptive effect of Casearia sylvestris extract in the acetic acid test was prevented by prior treatment with the non-selective opioid receptor antagonist naloxone (1mg/kg; 5.8+/-4.2 and 31.5+/-3.1 writhes in vehicle-treated and naloxone-treated groups, respectively), indicating that the endogenous opioid system is involved in its analgesic mechanism of action. Thus, our investigation suggests a potential therapeutic benefit of Casearia sylvestris Sw. in treating conditions associated with inflammatory pain. PMID- 17399926 TI - Fluorescent TiO2 powders prepared using a new perylene diimide dye: applications in latent fingermark detection. AB - A new, highly fluorescent dye was synthesised using oleylamine combined with a perylene dianhydride compound. The new dye was characterised by 1H NMR, UV-vis spectroscopy and fluorescence spectroscopy as well as quantum yield. The dye was absorbed onto titanium dioxide nanoparticles for use as a fingerprint detection powder. The new fluorescent powder was applied to latent fingermarks deposited onto different non-porous surfaces and compared with commercial fluorescent powders. The powder exhibits strong fluorescence at 650-700 nm under excitation at 505 nm. On glass surfaces, the new powder gave images showing tertiary-level detail of the fingermark ridges with almost no background development. Compared with current magnetic fluorescent powders, the new powder was slightly weaker in fluorescence intensity but produced significantly less background development, resulting in good contrast between the fingermark and the substrate. PMID- 17399927 TI - Immunohistochemical staining with cytokeratin combining semi-serial sections for detection of cervical lymph node metastases of oral squamous cell carcinoma. AB - OBJECTIVES: Lymphatic metastatic characteristics of oral squamous cell carcinoma are not fully understood, for instance, skip metastasis is still controversial. The purposes of the present study was to explore the accuracy and applicability of immunohistochemical stain with cytokeratin combining semi-serial sections for detection of cervical lymph node metastasis of oral squamous cell carcinoma. METHODS: Regional lymph nodes (N=1638) were obtained from 26 patients with primary oral squamous cell carcinoma who underwent five level neck dissections. Semi-serial sections at an interval of 0.5mm was performed for each lymph node and cross-detected by immunohistochemical staining with cytokeratin and traditional hematoxylin-eosin staining (H-E) and their accuracies were compared. RESULTS: Of 26 patients, 21 were detected having lymphatic metastasis by H-E staining and 26 by immunohistochemical detection; Of 1638 lymph nodes, 52 metastatic lymph nodes were detected by H-E staining while 162 by immunohistochemical detection. One case with cancer of the mouth floor being defined having skip metastasis was proved having no skip metastasis by the immunohistochemical detection. CONCLUSIONS: The immunohistochemical detection method with semi-serial sections has higher accuracy than the traditional H-E staining and its application may present a need to re-evaluate the neck metastatic patterns of oral squamous cell carcinoma. PMID- 17399928 TI - [Lyme disease: prophylaxis after tick bite]. AB - Lyme disease is a bacterial infection caused by Borrelia burgdorferi, which is transmitted by infected ticks. The transmission depends on several factors, especially on the duration of the tick's presence in the host body (the nymph which is smaller than the adults and thus less visible, is in this case the most frequently involved) and on whether the tick is infected or not. The interpretation of results in the few available studies is made difficult by the lack of information obtained (due to difficulty to collect information and examination costs). The comparison is made even more difficult by the difference between Borrelia ticks species in various regions. Today, the best methods are preventive: protective clothing, tick repellents, checking and removal of ticks after a journey in an endemic zone, and in case of tick bite, regular examination of the bite site during the following weeks in order to initiate an early curative treatment if ECM is diagnosed. The currently available data seems to be insufficient to suggest systematic antimicrobial prophylaxis in case of tick bite. PMID- 17399929 TI - Age-related changes of P2X(1) receptor mRNA in the bladder detrusor from men with and without bladder outlet obstruction. AB - The urinary bladder purinergic system is reported to change with age and with bladder dysfunction. Here, we examined the expression of purinergic P2X(1) receptors in detrusor and mucosa (urothelium+lamina propria) from male control bladder and investigated age-related P2X(1) receptor mRNA expression in control and obstructed detrusor. Biopsy specimens were obtained at cystoscopy from control patients (n=46, age range 30-86years) and patients diagnosed with outlet obstruction (n=29, 46-88years). Calponin expression (measured by RT-PCR) was similar in control and obstructed detrusor and did not change with age. Quantitative competitive RT-PCR was used to measure P2X(1) receptor and GAPDH mRNA in control and obstructed detrusor. P2X(1) receptor mRNA expression was 9 fold (p<0.0001) higher in the detrusor than in the mucosa. Expression of mRNA for the internal control GAPDH remained stable with age and across control and obstructed detrusor. No difference in P2X(1) receptor expression was observed between control and obstructed detrusor (p=0.35). However, an age-related decrease in P2X(1) mRNA expression was observed in control (n=27; p=0.0054; Spearman coefficient r=-0.520) but not obstructed detrusor (n=19; p=0.093; r= 0.396). Downregulation of P2X(1) mRNA expression might occur as a result of an increased component of neural ATP release in the aging bladder. PMID- 17399930 TI - Angiographic frequency of blunt cerebrovascular injury in patients with carotid canal or vertebral foramen fractures on multidetector CT. AB - PURPOSE: Blunt carotid injuries (BCI's) and blunt vertebral artery injuries (BVI's), known jointly as BCVI's, are common in "high risk" patients. The purpose is to evaluate the rate of occurrence of BCI/BVI in patients screened purely by the radiologic criteria of fracture through the carotid canal or vertebral transverse foramina, or significant cervical subluxation, noted by multidetector CT. METHODS: Seventy-one patients with 108 catheterized vessels were included over a 13-month interval. The angiographic examinations were prompted by current hospital protocol, solely by the presence of fractures involving/adjacent to the carotid canal, cervical fractures involving/adjacent to the foramen transversarium, or cervical fractures with significant subluxation. The incidence of each grade of blunt injury was calculated after review of the CT scans and catheter angiograms by two neuroradiologists. RESULTS: Two thousand and seventy three total blunt trauma admissions occurred during the time period, with a BCVI rate of 0.92-1.0% (depending on the reviewer), similar to previous studies. Mean time to catheter angiography was 16.6 h. Of the 71 included patients, there were 11-12 BCI's and 10-12 BVI's, an overall rate of 27-30% of BCVI in the patients with foraminal fractures. Interobserver agreement in reviewing the catheter angiograms was excellent (Kappa 0.795). Of note, three internal carotid pseudoaneurysms resolved spontaneously after anticoagulation or aspirin. CONCLUSION: This study confirms that there is a high rate of BCVI in the presence of carotid canal or vertebral foramen fractures that are noted by multidetector CT. Utilization of purely radiologic criteria of foraminal involvement may be a significant screening tool in the decision of whether to evaluate these patients acutely by catheter or CT angiography, and for early detection of patients at risk for symptomatology, to initiate prompt, prophylactic treatment. PMID- 17399932 TI - Clinical application of 64-slice spiral CT in the diagnosis of the Tetralogy of Fallot. AB - OBJECTIVE: To explore the clinical application of 64-slice spiral CT in the diagnosis of the Tetralogy of Fallot. METHODS: Eighteen patients with diagnosed Tetralogy of Fallot underwent cardiac CT angiography with 64-slice CT (CTA). Two- and three-dimensional images were used for diagnosing in all cases by means of MPR (coronal, sagital and oblique), cMPR, MIP and VRT. RESULTS: All patients had ventricular septal defect, pulmonary stenosis and hypertrophy of right ventricle. The morphologic features of Tetralogy of Fallot were equal to echocardiography. Patent ductus arteriosus (PDA) were detected on eight patients. Main coronary artery branches and partial sub-branches were visualized in all patients. Abnormal coronary arteries were found in seven cases: both left and right main coronary artery arising from the right aortic sinus one case, right main coronary artery and circumflex (CX) arising from the right aortic sinus one case, left anterior descending (LAD) and CX arising from left aortic sinus directly one case, left and right coronary artery arising from back and front of the aortic sinus, respectively, three cases. Pulmonary artery branch stenosis was found in 12 cases and branch pulmonary artery dilation noticed in 1 case. Double superior vena cava was also found in one case. According to the surgery the diagnostic accuracy of CT and Echocardiography was 95.45 and 83.33%, respectively. CONCLUSION: Two- and three-dimensional 64-slice CTA not only show the overall anatomical structure of the heart, but also show coronary and pulmonary arteries. With these results, evaluation of coronary anomalies and pulmonary artery stenosis with 64-MSCT is extremely valuable for planning the operative procedure on the patients with Tetralogy of Fallot. PMID- 17399933 TI - Differentiation of benign and malignant parotid tumors using deconvolution-based perfusion CT imaging: feasibility of the method and initial results. AB - AIM: We evaluated the feasibility of perfusion CT (CTP) of the parotid gland and attempted to differentiate benign from malignant tumors. MATERIALS AND METHODS: CTP was performed in 17 patients with benign tumors and 10 patients with malignant parotid tumors. Data were postprocessed by using deconvolution-based perfusion analysis. Postprocessing-generated maps showed blood flow (BF), blood volume (BV), mean transit time (MTT), and capillary permeability surface product (PS). Regions of interest were placed through the tumor site and the contralateral healthy parotid tissue. Ratios of the perfusion values between the tumors and the contralateral healthy structures were also calculated. Pearson correlation coefficients were determined to compare the agreement between the two readers. RESULTS: Perfusion maps of all tumors were successfully obtained. High Pearson correlation coefficients comparing the two readers' visually measured abnormalities were observed (r=0.79-0.86, P=0.001) for all perfusion maps, The MTT and PS values between malignant and benign tumors were not significantly different. The BF and BV values were statistically significant different between the benign and malignant tumors (0.00 or =15%, preoperative AFP > or =2000 ng/ml, and tumour reduction rate <98% were predictive of decreased overall survival. When the three prognostic factors were used in a scoring system, with one point assigned for each factor, the 3-year survival rates of patients with scores of 0, 1, 2, and 3 were 71%, 40%, 0%, and 0% respectively. CONCLUSIONS: Reduction hepatectomy followed by TACE is effective in patients with advanced T-Staged HCCs who have none of the 3 poor prognostic factors. Reduction surgery followed by TACE is one of the options for controlling advanced T-Staged HCCs in patients who are not candidates for curative resection or TACE alone. PMID- 17399941 TI - Efficacy and safety of oral beclomethasone dipropionate for ileal or ileal-right colon Crohn's disease of mild-to-moderate activity or in remission: Retrospective study. AB - Although conventional glucocorticosteroids are the main treatments for active Crohn's disease, several problems are associated with steroid dependence and steroid-related adverse events. To assess the efficacy and safety of oral beclomethasone dipropionate (BDP) coated tablets in adults with mild-to-moderate Crohn's disease. Thirty-four patients (age 18-70years) with a diagnosis of Crohn's disease confirmed by conventional criteria (barium enema, clinical criteria, colonoscopy, histology) were retrospectively evaluated in the study. All subjects received a treatment schedule with BDP 5-10mg/day for 24weeks. BDP significantly (p=0.005) reduced mean Crohn's Disease Activity Index (CDAI) score from 169.6 at baseline to 123.2 after 24weeks. Clinical success was evident at 24weeks in 66.7% of patients with initial active disease, and remission was maintained at week 24 in 93.8% of patients with remission at baseline. Overall, female non-smokers had the best response to treatment. BDP was well tolerated and the only adverse events observed were nausea (n=1), facial erythema (n=1) and one patient with raised fasting blood glucose level. These results clearly suggest that oral BDP coated tablets are effective and safe for treatment of mild-to moderate Crohn's disease of ileal or ileal-right colonic localisation. PMID- 17399942 TI - Characterization of cell death events induced by anti-neoplastic drugs cisplatin, paclitaxel and 5-fluorouracil on human hepatoma cell lines: Possible mechanisms of cell resistance. AB - Two different hepatoma cell lines were incubated for 48h with chemotherapeutic drugs cisplatin, paclitaxel and 5-FU to determine their ability to induce cytotoxicity and DNA fragmentation as well as to modify the expression of some cell death-related genes that could be involved in the resistance to therapy. We observed that cisplatin and paclitaxel induced cytotoxicity, but significant differences between both cell lines, were found only in the case of paclitaxel. At 48h, apoptosis was clearly present in Hep3B cells treated with cisplatin and HepG2 cells treated with paclitaxel. 5-FU induced cytotoxicity in both cell lines but only at higher concentrations than the other two drugs, triggering apoptosis and necrosis in HepG2 cells and only necrosis in Hep3B. When a time course was performed for the first 8h of treatment to elucidate the initial mechanism of cell death responsible for DNA fragmentation, we observed that 5-FU in Hep3B, and cisplatin in both cell lines, induces primary necrosis, whereas at the concentration tested here, paclitaxel clearly triggers apoptosis in both cell lines. HepG2 cells were weakly sensitive to 5-FU in the first 8h of treatment, so the primary mechanism of cell death was not clear, but results seem to indicate that it could be apoptosis. At 48h, Bax was not up-regulated with any of the treatments, whereas cisplatin was able to induce Bcl-xL down-regulation in both cell lines. Treatment with 5-FU also down-regulated Bcl-xL in HepG2 cells. We also measured variations in the expression of survivin, an inhibitor of apoptosis that has also been involved in mitototic catastrophe. Hep3B cells seem to show an increase in protein levels with all treatments. Exposure to paclitaxel resulted in the highest effect. In the case of HepG2 cells, there was a decrease in survivin expression when cells were treated with 5FU and paclitaxel, both treatments showing complete loss of the protein. Using an antibody that recognizes unprocessed caspase-3, we observed that the enzyme was assumingly activated in HepG2 cells treated with 5FU and paclitaxel, but only weakly after treatment with cisplatin. Hep3B cells did not show activation since the levels of the pro-enzyme remained the same as that in the control. In conclusion, the three drugs tested in this study could induce cell death, with paclitaxel being more effective inducing apoptosis. 5FU was only effective at high doses and its mechanism seems to be primarily related to necrosis in Hep3B and probably apoptosis in HepG2. Cisplatin mechanism of cell death is probably mediated by the decrease in anti-apoptotic protein Bcl-xL whereas paclitaxel and 5FU are decreasing the apoptosis inhibitor survivin. According to pro-enzyme levels, caspase-3 was only activated in HepG2 cells, whereas in the case of Hep3B cells the mechanisms of toxicity appear to be caspase-3-independent at the time and concentrations tested in this study. The resistance of Hep3B cells to death induced by chemotherapy could be related to an increase in the expression of IAP survivin, which can decrease cell response to the treatment or even switch the type of death from apoptosis to another kind, making therapy less efficient. PMID- 17399943 TI - [Shigellosis or bacillary dysentery]. AB - Shigellosis, commonly known as bacillary dysentery, is an enterobacterial disease caused by the Shigella genus, which now belongs to the Escherichia tribe, because of their genetic and phenotypic similarities. S. sonnei, flexneri, boydii and dysenteriae differ in their epidemiologic and pathogenic characteristics. S. sonnei is predominant in industrialized countries and causes the mildest disease. S. dysentery is especially dangerous because it occurs in outbreaks, sometimes in catastrophic situations, and has a high mortality rate. Shigellosis typically causes watery diarrhea followed by dysentery. It may be associated with a variety of mild to severe or even life-threatening complications. Shigellosis is a significant public health problem in developing countries where it remains a major cause of diarrhea-related morbidity and mortality, especially among children. Annual cases worldwide are estimated at 160 million. Shigellosis is a highly contagious disease of poor and crowded communities, with fecal-oral (hand to-mouth) transmission, and an extremely low minimum infectious dose. Multidrug resistance is a serious problem. Ciprofloxacin is the first-line antibiotic, recommended for 3 days. Prevention and control should include personal hygiene, hand washing, sanitation, and water supply. No vaccine is available, but several candidates are currently being evaluated. PMID- 17399944 TI - [Paraneoplastic neurological syndromes]. AB - The classic paraneoplastic neurological syndromes include Lambert-Eaton myasthenic syndrome, limbic encephalitis, sensory neuronopathy, intestinal pseudo obstruction, subacute cerebellar degeneration, encephalomyelitis, and dermatomyositis. Approximately ten onconeural antibodies that recognize cancer and the nervous system have been described in paraneoplastic neurological syndromes. These antibodies appear to be important diagnostic tools, even though they may not always be present. Deciding whether a given neurological picture is definitely or possibly paraneoplastic depends on the clinical syndrome, any association with onconeural antibodies, and the time elapsed between onset of neurological symptoms and the discovery of the cancer. Diagnosis of a classic paraneoplastic neurological syndrome or the discovery of onconeural antibodies mandates an active and persistent search for cancer, using new techniques such as fluorodeoxyglucose positron emission tomography. In patients with one of these syndromes, the best treatment of the neurological disease is often the diagnosis and early treatment of the cancer. PMID- 17399945 TI - Distribution of 14C-atrazine following an acute lactational exposure in the Wistar rat. AB - The purpose of the present study was to examine the distribution of atrazine in the lactating dam and suckling neonate following an acute exposure to either 2 or 4mg/kg 14C-atrazine (14C-ATR) by gavage. 14C-ATR was administered to the nursing dam on postnatal day 3 by oral gavage. Two and a half hours after exposure of the mother to 14C-ATR, the pups were allowed to nurse for 30min. At the end of the nursing period, radiolabelled residues of 14C-ATR [or 14C-chlorotriazines (14C ClTRI)] were measured in the organs and tissues of the perfused dam and in the stomachs and brains of the rat pups. Both the 2 and the 4mg atrazine treatments resulted in a transfer of approximately 0.007% of 14C-ClTRI to the stomach (indicator of milk content) and 0.0002% to the brains of the offspring following the 30-min nursing period. Three hours following the dose of 14C-ATR, there was a distribution of 14C-ClTRI to the organs of the dam, with the highest amounts in the liver and kidney (1.1 and 0.3% of the administered dose, respectively). Approximately 0.003% of the administered dose was present in three different brain sections of the dam following both doses of 14C-ATR. The results of this study demonstrate that 14C-ClTRI are present in small concentrations in the brain and tissues of the dam (adult female) and provide evidence that atrazine or the metabolites can have direct effects on neuroendrocrine function. The results also provide information for postnatal distribution into the suckling neonate during early lactation. PMID- 17399946 TI - Favorable pregnancy outcome following Trastuzumab (Herceptin) use during pregnancy--Case report and updated literature review. AB - BACKGROUND: Trastuzumab (Herceptin) is a monoclonal antibody used for the treatment of breast cancer. Experience with use of this agent during pregnancy, and its possible effects on the fetus, is limited. CASE: We present a case of a patient with breast cancer who was treated with trastuzumab during the first 24 weeks of pregnancy. This treatment was associated with reversible maternal heart failure, which resolved slowly after the drug was discontinued, but with no adverse fetal effects and a normal infant examination at the age of 2 months. Updated literature review is discussed. PMID- 17399947 TI - Optimal decision network with distributed representation. AB - On the basis of detailed analysis of reaction times and neurophysiological data from tasks involving choice, it has been proposed that the brain implements an optimal statistical test during simple perceptual decisions. It has been shown recently how this optimal test can be implemented in biologically plausible models of decision networks, but this analysis was restricted to very simplified localist models which include abstract units describing activity of whole cell assemblies rather than individual neurons. This paper derives the optimal parameters in a model of a decision network including individual neurons, in which the alternatives are represented by distributed patterns of neuronal activity. It is also shown how the optimal weights in the decision network can be learnt via iterative rules using information accessible for individual synapses. Simulations demonstrate that the network with the optimal synaptic weights achieves better performance and matches fundamental behavioural regularities observed in choice tasks (Hick's law and the relationship between the error rate and the time for decision) better than a network with synaptic weights set according to a standard Hebb rule. PMID- 17399948 TI - Power-law models of signal transduction pathways. AB - The mathematical modelling of signal transduction pathways has become a valuable aid to understanding the complex interactions involved in intracellular signalling mechanisms. An important aspect of the mathematical modelling process is the selection of the model type and structure. Until recently, the convention has been to use a standard kinetic model, often with the Michaelis-Menten steady state assumption. However this model form, although valuable, is only one of a number of choices, and the aim of this article is to consider the mathematical structure and essential features of an alternative model form--the power-law model. Specifically, we analyse how power-law models can be applied to increase our understanding of signal transduction pathways when there may be limited prior information. We distinguish between two kinds of power law models: a) Detailed power-law models, as a tool for investigating pathways when the structure of protein-protein interactions is completely known, and; b) Simplified power-law models, for the analysis of systems with incomplete structural information or insufficient quantitative data for generating detailed models. If sufficient data of high quality are available, the advantage of detailed power-law models is that they are more realistic representations of non-homogenous or crowded cellular environments. The advantages of the simplified power-law model formulation are illustrated using some case studies in cell signalling. In particular, the investigation on the effects of signal inhibition and feedback loops and the validation of structural hypotheses are discussed. PMID- 17399949 TI - Abl functions as a negative regulator of Met-induced cell motility via phosphorylation of the adapter protein CrkII. AB - HGF, the ligand for the Met receptor tyrosine kinase, is a potent modulator of epithelial-mesenchymal transition and dispersal of epithelial cells, which are processes that play a crucial role in cell motility during normal development and malignant transformation. We and others have shown earlier that the adapter protein CrkII and its associated proteins positively regulate cell migratory events in response to both haptotactic and chemotactic stimuli, including HGF. Here, we demonstrate for the first time that phosphorylation of CrkII serves as a negative feedback loop to regulate motile responses upon Met stimulation. Thus, we found that the treatment of cells with HGF induces tyrosine phosphorylation of CrkII at Y221, which in turn results in inhibition of CrkII signaling via formation of an intramolecular pY221-SH2-domain interaction. Accordingly, expression of a mutant form of CrkII, CrkII-Y221F, which is resistant to phosphorylation at this negative regulatory site, enhanced Met-induced cell motility. Furthermore, we demonstrate here that the Met-induced CrkII phosphorylation depends on the Abl tyrosine kinase activity. As a corollary, we found that Abl inhibitors, such as the STI571 compound, significantly enhanced Met-induced cell motility, but failed to do so in cells that expressed the CrkII Y221F mutant protein. Taken together, these results demonstrate that the Abl tyrosine kinase functions as a negative regulator of Met-induced cell migration, and that it does so by inducing CrkII phosphorylation at the site Y221. PMID- 17399950 TI - The evolution of preimplant surgery from preprosthetic surgery. PMID- 17399951 TI - CNS 14-3-3zeta: changes with sex but not psychiatric diagnoses or psychotropic drug treatment. AB - mRNA for 14-3-3zeta, an abundant signalling protein in human CNS, is reported as decreased or unchanged in cortex from subjects with schizophrenia. Addressing this dichotomy, using Western blot analyses, we measured levels of 14-3-3zeta proteins in cortex and caudate nucleus from subjects with schizophrenia, bipolar disorder, age/sex matched controls and in analogous CNS regions from rats treated with psychotropic drugs. Anti-14-3-3zeta antibody bound to three proteins (molecular weights: 27, 54 and 70 kDa), in all CNS tissue. Levels of all proteins did not vary with diagnoses (27 kDa: F(2,42.0)=0.35, p=0.71; 54 kDa: F(2,42.1)=0.62, p=0.54; 70 kDa: F(2,41.0)=2.43, p=0.10). By contrast, independent of diagnoses, there were significant increases in the levels of the 27 kDa protein (+32%; p<0.001) and 54 kDa protein (51%; p=0.001) in the caudate nucleus from males compared to females. In addition, there was a trend (-25%; p=0.06) to decreased levels of the 70 kDa protein in BA 9 in males compared to females. Treating with haloperidol, olanzapine, lithium or a combination thereof did not alter 14-3-3zeta levels in rat cortex or striatum. Therefore, this study suggests that 14-3-3zeta proteins are not altered in the cortex or caudate nucleus in schizophrenia, bipolar disorder or in analogous regions in psychotropic drug treated rats. By contrast, our study suggests that levels of 14-3-3zeta in some regions of the human CNS may be modulated by some sex-specific mechanism. PMID- 17399952 TI - Response inhibition and cognitive flexibility in schizophrenia with and without comorbid substance use disorder. AB - Addiction is a frequent comorbid disorder in schizophrenia related to dopaminergic dysfunction in fronto-subcortical circuits. These brain networks are relevant for both (executive) cognition and the neuropathology of schizophrenia. The aim of the present study was to elucidate whether response inhibition and cognitive flexibility - executive abilities relevant for achieving and maintaining abstinence - are differentially impaired in schizophrenia patients with or without comorbid substance use disorder. Patients suffering from major depression or alcoholism as well as healthy controls served as comparison groups. The ability to inhibit predominant response tendencies during response conflict and to efficiently shift the focus of attention between different task requirements was assessed by verbal and non-verbal cognitive tasks. Contrary to expectation, non-addicted schizophrenia patients showed the most pronounced executive function impairments relative to the control groups, affecting both response suppression and cognitive flexibility. Dual diagnosis patients did not differ significantly from non-addicted schizophrenia patients or from the alcoholic group, but were impaired at cognitive flexibility relative to the depression subgroup and healthy controls. Whether the relative preservation of response inhibition and cognitive flexibility in the dual disorder patients is due to high premorbid functioning, beneficial self-medication effects or compensatory brain activation remains to be elucidated. The relatively intact executive abilities in young, addicted schizophrenia patients might represent a beneficial resource for treatment strategies. PMID- 17399953 TI - Schizotypy versus openness and intelligence as predictors of creativity. AB - Schizophrenia-spectrum risk alleles may persist in the population, despite their reproductive costs in individuals with schizophrenia, through the possible creativity benefits of mild schizotypy in non-psychotic relatives. To assess this creativity-benefit model, we measured creativity (using 6 verbal and 8 drawing tasks), schizotypy, Big Five personality traits, and general intelligence in 225 University of New Mexico students. Multiple regression analyses showed that openness and intelligence, but not schizotypy, predicted reliable observer ratings of verbal and drawing creativity. Thus, the 'madness-creativity' link seems mediated by the personality trait of openness, and standard creativity benefit models seem unlikely to explain schizophrenia's evolutionary persistence. PMID- 17399954 TI - Anterior cingulate volumes in schizophrenia: a systematic review and a meta analysis of MRI studies. AB - OBJECTIVES: Several MRI studies have investigated the anterior cingulate in schizophrenia, as this is a key region for emotional processing and higher executive performances. A systematic review of structural MRI studies and a meta analysis were conducted to explore whether anterior cingulate volumes are abnormal in patients with schizophrenia. METHOD: A systematic search strategy was used to identify eligible MRI studies. Thereafter, a meta-analysis was carried out by using a random effect model. Also, a meta-regression analysis was used to assess the influence of age, gender and slice thickness on effect sizes. RESULTS: The meta-analysis was performed on seven studies. These results showed that the anterior cingulate volumes were significantly reduced in patients compared to healthy controls. Significant heterogeneity between these studies was observed. The meta-regression demonstrated that the effect size was significantly related only to slice thickness. CONCLUSIONS: Our work confirmed the presence of abnormally reduced anterior cingulate volumes in schizophrenia. However, several methodological issues limited the interpretation of these findings. Among these were different MR acquisition parameters and the small size of the sample, which was mostly composed of chronic patients. Future MRI studies should be planned to better understand the functional expression of anterior cingulate structural abnormalities. PMID- 17399955 TI - The up-regulation of angiotensin II receptor type 1 and connective tissue growth factor are involved in high-glucose-induced fibronectin production by cultured human dermal fibroblasts. AB - BACKGROUND: The expansion of extracellular matrix in diabetes occur in many tissues, including skin, but the underlying mechanisms are poorly understood. We were interested to study whether the activation of angiotensin II/receptor type 1 pathway with the consequent involvement of CTGF may be the possible cause of high glucose-induced matrix abnormalities in cultured dermal fibroblasts. OBJECTIVE: We aimed to investigate the effect of high glucose on the generation of angiotensin II and the expression of angiotensin II receptors, and on the expression of CTGF mediating the fibronectin production in cultured human dermal fibroblasts. METHODS: Cell culturing, ELISA, semi-quantitative RT-PCR, Western blotting. RESULTS: High glucose treatment of cultured dermal fibroblasts led to: (1) the angiotensin II receptor type 1 was up-regulated at the level of mRNA and protein, unlike the receptor type 2; (2) the generation of angiotensin II and the mRNA expression of all components of the local renin-angiotensin system were not altered; (3) the mRNA and protein expression of CTGF was up-regulated, and this effect was cancelled by losartan; (4) the fibronectin production was increased, also was cancelled by losartan, while an anti-CTGF-neutralizing antibody only partly reduced it; (5) TGFbeta1 expression, the secretion of total and active TGFbeta1 were not changed; (6) the hyperosmotic action of high glucose had no effect. CONCLUSION: The up-regulation of angiotensin II receptor type 1 and the consequent increase of CTGF expression, independently of TGFbeta1, participate in high-glucose-induced fibronectin production in cultured human dermal fibroblasts. PMID- 17399956 TI - Biofilm formation by Propionibacterium acnes is associated with increased resistance to antimicrobial agents and increased production of putative virulence factors. AB - Propionibacterium acnes plays an important role in the pathogenesis of acne vulgaris, a common disorder of the pilosebaceous follicles. Recently, it was suggested that P. acnes cells residing within the follicles grow as a biofilm. In the present study, we tested the biofilm-forming ability of several P. acnes strains in a microtiter plate model. We also evaluated the resistance of biofilm grown P. acnes towards antimicrobial agents commonly used in the treatment of acne and the production of putative virulence factors. Our results indicate that P. acnes can form biofilms in vitro. The results also show that sessile P. acnes cells are more resistant to various commonly used antimicrobial agents than planktonic cells. In addition, sessile cells produce more extracellular lipases as well as significant amounts of the quorum-sensing molecule autoinducer-2. PMID- 17399957 TI - Quinoline alkaloids from Lunasia amara inhibit Mycobacterium tuberculosis H37Rv in vitro. PMID- 17399959 TI - Fronto-temporal-lobe atrophy in early-stage Alzheimer's disease identified using an improved detection methodology. AB - Alzheimer's disease (AD) is associated with widespread brain atrophy including structures subserving memory. We applied an improved structural detection methodology to examine the less well known progression of atrophy in early-stage AD. We sought to i) longitudinally study volumetric differences in patients with early-stage AD and healthy volunteers; and ii) test the hypothesis that hippocampal volumes would be correlated with clinically relevant cognitive function. Seven patients and eleven healthy subjects underwent two structural MRI scans and neuropsychological assessments. Scans were normalised to a study specific template and 'morphologically opened' to reduce tissue misclassification. Using brain-parcellation, patient atrophy was localised to left fusiform and parahippocampal gyri, whilst left hippocampal volumes were correlated with a cognitive performance measure. A whole-brain search methodology, showed that patients had reduced volumes including fronto-temporal regions bilaterally, in hippocampi and amygdalae and right cerebellum. Whole brain correlational analyses revealed that cognitive performance was correlated with volumes of both hippocampi, superior temporal gyri and left insula. Neither group exhibited significant longitudinal volumetric changes. Utilising a novel methodology, we have shown that in early-stage AD, clinically relevant cognitive deficits are correlated with regionally specific grey-matter volumes, which are detectable at an early stage of the illness. PMID- 17399958 TI - TRP channels in hypertension. AB - Pulmonary and systemic arterial hypertension are associated with profound alterations in Ca(2+) homeostasis and smooth muscle cell proliferation. A novel class of non-selective cation channels, the transient receptor potential (TRP) channels, have emerged at the forefront of research into hypertensive disease states. TRP channels are identified as molecular correlates for receptor-operated and store-operated cation channels in the vasculature. Over 10 TRP isoforms are identified at the mRNA and protein expression levels in the vasculature. Current research implicates upregulation of specific TRP isoforms to be associated with increased Ca(2+) influx, characteristic of vasoconstriction and vascular smooth muscle cell proliferation. TRP channels are implicated as Ca(2+) entry pathways in pulmonary hypertension and essential hypertension. Caveolae have recently emerged as membrane microdomains in which TRP channels may be co-localized with the endoplasmic reticulum in both smooth muscle and endothelial cells. Such enhanced expression and function of TRP channels and their localization in caveolae in pathophysiological hypertensive disease states highlights their importance as potential targets for pharmacological intervention. PMID- 17399960 TI - Protein adsorption characteristics of calcium hydroxyapatites modified with pyrophosphoric acids. AB - Protein adsorption characteristics of calcium hydroxyapatite (Hap) modified with pyrophosphoric acids (PP(a)) were examined. The PP(a) modified Hap particles (abbreviated as PP-Hap) possessed anchored polyphosphate (PP: P-{O-PO(OH)}(n)-OH) branches on their surfaces. The proteins of bovine serum albumin (BSA: isoelectric point (iep)=4.7, molecular mass (M(s))=67,200 Da, acidic protein), myoglobin (MGB: iep=7.0, M(s)=17,800 Da, neutral protein), and lysozyme (LSZ: iep=11.1, M(s)=14,600 Da, basic protein) were examined. The zeta potential (zp) of PP-Hap particles as a function of pH overlapped; zp-pH curves were independent of the concentration of pyrophosphoric acids (abbreviated as [PP(a)]) used for modifying Hap surface. The saturated amounts of adsorbed BSA (Delta n(ads)(BSA)) were increased three-fold by the surface modification with PP(a) though they were independent of the [PP(a)]. Furthermore, the fraction of BSA desorption was independent of the [PP(a)]. This enhancement of BSA adsorption onto the PP-Hap is due to the hydrogen bonding between oxygen and OH groups of the PP-branches and functional groups of BSA molecules. In the case of LSZ, a more higher adsorption enhancement was observed; the saturated amount of adsorbed LSZ (Delta n(ads)(LSZ)) for Hap modified at [PP(a)]=6 mmol/dm(3) was nine-fold than that for Hap unmodified. This remarkable adsorption enhancement was explained by a three dimensional binding mechanism; LSZ molecules were trapped inside of the PP branches. Hence, a fraction of LSZ desorption was decreased with an increase in the [PP(a)]; as more PP-branches are presented on the surface the higher retardation of LSZ desorption was induced. It was expected from their small size that MGB adsorb between the PP-branches as well as LSZ. However, the amounts of adsorbed MGB (Delta n(ads)(MGB)) did not vary and were independent of the [PP(a)] due to the small numbers of functional groups of MGB. In addition, no dependence of the fraction of MGB desorption on the [PP(a)] was observed. The results of zp for all the protein systems supported the mode of protein adsorption discussed. The anchored structure of the PP-branches developed on the Hap surface to provide three-dimensional protein adsorption spaces was proved by a comparative experiment that was elucidating the effect of pyrophosphate ions for BSA adsorption onto Hap. PMID- 17399961 TI - Stability of S-layer proteins for electrochemical nanofabrication. AB - Crystalline cell surface layer proteins (S-layers) can be used in electrochemical fabrication to create nanoscale arrays of metals and oxides on surfaces so long as the proteins maintain their long-range order during processing. We have explored the stability of the HPI layer protein (the S-layer protein from the microorganism Deinococcus radiodurans) adsorbed onto platinum surfaces after immersion in sulfuric acid or sodium hydroxide electrolytes ranging in pH from 0 to 14 over time periods ranging from 1 to 1000s. Topographic data obtained by atomic force microscopy (AFM) was used to characterize the protein stability, judged by its retention of long-range order after immersion. The compiled data revealed that, under these solution conditions and in this environment, the HPI layer protein has a dose-dependent structural stability "envelope" in the acidic range from 110 weeks). PCR analysis showed that the isolates carried various virulence genes such as Ehly, eae, stx1 and stx2, which highlight the potential importance of these attributes for the infection, colonization and the possible pathogenesis of calf diarrhea. Cytotoxicity analysis revealed that many of the EHEC isolates showed high cytotoxicity to Vero cells, re-emphasizing the potential for cattle being a direct source of EHEC infections in humans. PMID- 17400009 TI - The cerebellum and cognition. AB - The most important function of the cerebellum may be to coordinate motor function so that movements can be performed smoothly, but there are others. It has been shown that the cerebellum is involved in certain aspects of cognition and changes in affect. Also verbal deficits can be found after cerebellar lesions. The cerebellar cognitive affective syndrome is described, and the evidence for its existence discussed; in particular the use of neuroimaging studies. Different areas of the cerebellum have been identified as serving the various functions, and also their connections to the relevant parts of the cerebral cortex. Certain conditions merit special attention. The function of spatial navigation needs a major contribution from the cerebellum, and the problems of autism and impaired cognition are no doubt related to the enlarged cerebellum described in this disorder. The cognitive defects found in children with cerebellar ataxia supports its role in learning, and so does the study of music. PMID- 17400010 TI - Tailored slice selection in solid-state MRI by DANTE under magic-echo line narrowing. AB - We propose a method of slice selection in solid-state MRI by combining DANTE selective excitation with magic-echo (ME) line narrowing. The DANTE RF pulses applied at the ME peaks practically do not interfere with the ME line narrowing in the combined ME DANTE sequence. This allows straightforward tailoring of the slice profile simply by introducing an appropriate modulation, such as a sinc modulation, into the flip angles of the applied DANTE RF pulses. The utility of the method has been demonstrated by preliminary experiments performed on a test sample of adamantane. PMID- 17400011 TI - A new detection scheme for ultrafast 2D J-resolved spectroscopy. AB - Recent ultrafast techniques enable 2D NMR spectra to be obtained in a single scan. A modification of the detection scheme involved in this technique is proposed, permitting the achievement of 2D 1H J-resolved spectra in 500 ms. The detection gradient echoes are substituted by spin echoes to obtain spectra where the coupling constants are encoded along the direct nu2 domain. The use of this new J-resolved detection block after continuous phase-encoding excitation schemes is discussed in terms of resolution and sensitivity. J-resolved spectra obtained on cinnamic acid and 3-ethyl bromopropionate are presented, revealing the expected 2D J-patterns with coupling constants as small as 2 Hz. PMID- 17400012 TI - Cytotoxic activity of a dichloromethane extract and fractions obtained from Eudistoma vannamei (Tunicata: Ascidiacea). AB - This study consists of the bioassay-guided fractionation of the dichloromethane extract from Eudistoma vannamei and the pharmacological characterization of the active fractions. The dried hydromethanolic extract dissolved in aqueous methanol was partitioned with dichloromethane and chromatographed on a silica gel flash column. The anti-proliferative effect was monitored by the MTT assay. Four of the latest fractions, numbered 14 to 17, which held many chemical similarities amongst each other, were found to be the most active. The selected fractions were tested for viability, proliferation and death induction on cultures of HL-60 promyeloblastic leukemia cells. The results suggested that the observed cytotoxicity is related to apoptosis induction. PMID- 17400013 TI - Plumage carotenoids of the Pin-tailed Manakin (Ilicura militaris): evidence for the endogenous production of rhodoxanthin from a colour variant. AB - The Pin-tailed Manakin (Ilicura militaris) is a small, sexually dimorphic, frugivorous suboscine songbird (Pipridae; Passeriformes; Aves) endemic to the Atlantic Forest of Brazil. A variant individual of this species was recently described in which the red patches that characterise the male's Definitive plumage were replaced by orange-yellow ones. We show here that the pigments in the feathers of the colour variant are common dietary carotenoids (zeaxanthin, beta-cryptoxanthin), not carotenoids synthesised by birds, lending support to the suggestion that the individual is a colour mutant lacking the capability to transform yellow dietary pigments into the red pigments normally present in these feathers. By comparison, the yellow crown feathers of a close relative, the Golden-winged Manakin (Masius chrysopterus), contained predominantly endogenously produced epsilon-caroten-3'-ones. Surprisingly, the normal-coloured feathers of the male Pin-tailed Manakin owe their red hue to rhodoxanthin, an unusual carotenoid more commonly found in plants, rather than 4-keto-carotenoids typically found in red plumages and found lacking in previously characterised bird colour variants. The implication is that birds, like the tilapia fish, may be able to synthesise this unusual pigment endogenously from dietary precursors. A newly described carotenoid, 6-hydroxy-epsilon,epsilon-carotene-3,3'-dione, here named piprixanthin, present in the red feathers of the Pin-tailed Manakin, provides a plausible intermediate between epsilon,epsilon-carotene-3,3'-dione (canary-xanthophyll B), a bright yellow pigment found in this and other songbirds, and rhodoxanthin. It is apparent that pigeons (Columbidae, Columbiformes) also have the capability to produce rhodoxanthin, and a structurally related pigment, endogenously. The ability to synthesise rhodoxanthin might have arisen at least twice in birds. PMID- 17400014 TI - Prostaglandin E2 inhibits Paracoccidioides brasiliensis killing by human monocytes. AB - Human monocytes lacked fungicidal activity against high virulence strain of Paracoccidioides brasiliensis, even after IFN-gamma activation. However, monocytes treated with indomethacin exhibited an effective killing against this fungus, suggesting a role of prostaglandin E2 (PGE2) in the inhibition process. Thus, the purpose of this work was to determine whether the effect of PGE2 in fungicidal activity was related with decrease on H(2)O(2) release, the metabolite involved in P. brasiliensis killing, and changes in the levels of TNF-alpha, IL-6 and IL-10. Human monocytes challenged with the fungus produced high PGE2 levels, which in turn inhibited the fungicidal activity of these cells by reducing H(2)O(2) and TNF-alpha production. PMID- 17400015 TI - Adoptive transfer of dendritic cells pulsed with Leishmania infantum nucleosomal histones confers protection against cutaneous leishmaniosis in BALB/c mice. AB - The mechanisms underlying the protective effects induced by dendritic cells (DC) based vaccines against Leishmania major in mice are not yet completely understood. In the present study, we investigated the potential of DC loaded with a mixture of the Leishmania infantum histones in the absence (HIS-pulsed DC) or presence of CpG motifs (HIS+CpG-pulsed DC) as a candidate vaccine against cutaneous leishmaniosis. Our data showed that a single intravenous administration of HIS-pulsed DC or HIS+CpG-pulsed DC confers control to L. major infection in BALB/c mice. Interestingly, all HIS-pulsed DC vaccinated mice remained susceptible to a second challenge. We found that the efficient immunity in BALB/c mice was associated to a Th1 response and a restriction of Th2 type of response upon challenge with L. major parasites. More importantly, the anti-leishmanial immunological mechanisms of protection were dependent on the ability to induce a low frequency of Foxp3(+) regulatory T cells at the site of infection. These results document that a vaccine based on a HIS+CpG-pulsed DC formulation may be as efficient for vaccination as one based on L. major antigen (Lm)+CpG-pulsed DC. Thus, HIS+CpG-pulsed DC may prove to be a new and further tool to add to those designed. PMID- 17400016 TI - Streptococcus agalactiae invasion of human brain microvascular endothelial cells is promoted by the laminin-binding protein Lmb. AB - Streptococcus agalactiae (S. agalactiae) can cause severe pneumonia, sepsis and meningitis in neonates and remains one of the most prevalent causes of invasive neonatal infections. During the course of infection, S. agalactiae colonizes and invades a number of host compartments, thereby interacting with different host tissues. Deletion of the scpB-lmb region, coding for the C5a peptidase and the laminin-binding protein Lmb, respectively, resulted in a 64% decreased invasion of S. agalactiae into human brain microvascular endothelial cells (HBMEC). Decreased invasion was also seen in lmb mutant strains lmb-k1 and lmb-k2 (74% and 69% reduction, respectively). Finally, host cell invasion was significantly reduced in competition experiments with either purified recombinant laminin binding protein by 46% or a polyclonal antibody directed against the laminin binding protein of S. agalactiae by 45%. The S. agalactiae scpB-lmb mutant induced an equal amount of the neutrophil chemoattractant interleukin (IL)-8 release in comparison to the wild-type. Taken together, our studies support the conclusion that Lmb promotes invasion of S. agalactiae into HBMEC but does not play a role in IL-8 release from HBMEC. PMID- 17400017 TI - Optimization of ultrasound extraction of phenolic compounds from coconut (Cocos nucifera) shell powder by response surface methodology. AB - Coconut is a tropical fruit largely consumed in many countries. In some areas of the Brazilian coast, coconut shell represents more than 60% of the domestic waste volume. The coconut shell is composed mainly of lignin and cellulose, having a chemical composition very similar to wood and suitable for phenolic extraction. In this work, the use of ultrasound to extract phenolic compounds from coconut shell was evaluated. The effect of temperature, solution to solid ratio, pH and extraction time were evaluated through a 2(4) experimental planning. The extraction process was also optimized using surface response methodology. At the optimum operating condition (30 degrees C, solution to solid ratio of 50, 15 min of extraction and pH 6.5) the process yielded 22.44 mg of phenolic compounds per gram of coconut shell. PMID- 17400018 TI - 2D modeling and preliminary in vitro investigation of a prototype high gradient magnetic separator for biomedical applications. AB - High gradient magnetic separation (HGMS) of magnetic materials from fluids or waste products has many established industrial applications. However, there is currently no technology employing HGMS for ex-vivo biomedical applications, such as for the removal of magnetic drug- or toxin-loaded spheres from the human blood stream. Importantly, human HGMS applications require special design modifications as, in contrast to conventional use where magnetic elements are permanently imbedded within the separation chambers, medical separators need to avoid direct contact between the magnetic materials and blood to reduce the risk of blood clotting and to facilitate convenient and safe treatment access for many individuals. We describe and investigate the performance of a magnetic separator prototype designed for biomedical applications. First, the capture efficiency of a prototype HGMS separator unit consisting of a short tubing segment and two opposing magnetizable fine wires along the outside of the tubing was investigated using 2D mathematical modeling. Second, the first-pass effectiveness to remove commercially available, magnetic polystyrene spheres from human blood using a single separator unit was experimentally verified. The theoretical and experimental data correlated well at low flow velocities (<5.0 cm/s) and high external magnetic fields (>0.05 T). This prototype separator unit removed >90% in a single pass of the magnetic spheres from water at mean flow velocity < or =8.0 cm/s and from blood mimic fluids (ethylene glycol-water solutions) at mean flow velocity < or =2.0 cm/s. In summary, we describe and prove the feasibility of a HGMS separator for biomedical applications. PMID- 17400019 TI - Clinical profile of various sub-types of writer's cramp. AB - We profiled 141 consecutive patients of writer's cramp (WC). Simple WC (n=108) had a mean (SD) disease duration of 3.65 (2.7) years and a severity score of 2.8 (0.91) by Burke-Fahn-Marden (BFM) scale (handwriting score: 0-4 ), whereas complex WC (n=33) had a disease duration of 5.8 (2.81) years and severity score of 3.6 (0.52). Compared with focal WC, generalized WC cases had longer disease duration, severer disability, and were older. Mirror dystonia was seen in 65 patients (46.1%). Co-existent Meige's syndrome (n=4) and cervical dystonia (n=6) were seen. Some patients had abnormal posturing of the neck (n=5) and of the oromandibular region (n=3) only while writing. Those receiving botulinum toxin fared significantly better than those on oral therapy, at 1 month post treatment. PMID- 17400020 TI - A review of plantar heel pain of neural origin: differential diagnosis and management. AB - Plantar heel pain is a symptom commonly encountered by clinicians. Several conditions such as plantar fasciitis, calcaneal fracture, rupture of the plantar fascia and atrophy of the heel fat pad may lead to plantar heel pain. Injury to the tibial nerve and its branches in the tarsal tunnel and in the foot is also a common cause. Entrapment of these nerves may play a role in both the early phases of plantar heel pain and recalcitrant cases. Although the contribution of nerve entrapment to plantar heel pain has been well documented in the literature, its pathophysiology, diagnosis and management are still controversial. Therefore, the purpose of this article was to critically review the available literature on plantar heel pain of neural origin. Possible sites of nerve entrapment, effectiveness of diagnostic clinical tests and electrodiagnostic tests, differential diagnoses for plantar heel pain, and conservative and surgical treatment will be discussed. PMID- 17400021 TI - Hydatid disease of the parotid gland: a rare case report. AB - A 40-year-old female suffering from hydatid disease located in the parotid gland is presented. Although Greece remains an endemic area for echinococcosis, this presentation of the disease is rare. Total excision of the cyst with partial parotidectomy was performed. The patient refused to receive general anesthesia and the operation was carried out under local anesthesia. Perioperative adjuvant medical therapy with albendazole was administered. In a two-year follow-up no recurrence has occurred. PMID- 17400022 TI - Peroxisome proliferator-activated receptors (PPARs) in skin health, repair and disease. AB - Peroxisome proliferator-activated receptors, PPARalpha, PPARbeta/delta and PPARgamma, are fatty acid activated transcription factors that belong to the nuclear hormone receptor family. While they are best known as transcriptional regulators of lipid and glucose metabolism, evidence has also accumulated for their importance in skin homeostasis. The three PPAR isotypes are expressed in rodent and human skin. Various cell culture and in vivo approaches suggest that PPARalpha contributes to fetal skin development, to epidermal barrier maturation and to sebocyte activity. PPARbeta/delta regulates sebocyte differentiation, promotes hair follicle growth and has pro-differentiating effects in keratinocytes in normal and inflammatory conditions. In contrast, the role of PPARgamma appears to be rather minor in keratinocytes, whereas its activity is required for sebaceous gland differentiation. Importantly, PPARalpha and beta/delta are instrumental in skin repair after an injury, each of them playing specific roles. Due to their collective diverse functions in skin biology, PPARs represent a major research target for the understanding and treatment of many skin diseases, such as benign epidermal tumors, papillomas, acne vulgaris and psoriasis. PMID- 17400023 TI - The present state of species within the genera Streptococcus and Enterococcus. PMID- 17400024 TI - Internal phthalate exposure over the last two decades--a retrospective human biomonitoring study. AB - In a retrospective human biomonitoring study we analyzed 24h urine samples taken from the German Environmental Specimen Bank for Human Tissues (ESBHum), which were collected from 634 subjects (predominantly students, age range 20-29 years, 326 females, 308 males) in 9 years between 1988 and 2003 (each n >or= 60), for the concentrations of primary and/or secondary metabolites of di-n-butyl phthalate (DnBP), di-iso-butyl phthalate (DiBP), butylbenzyl phthalate (BBzP), di(2-ethylhexyl) phthalate (DEHP) and di-iso-nonyl phthalate (DiNP). Based on the urinary metabolite excretion we estimated daily intakes of the parent phthalates and investigated the chronological course of the phthalate exposure. In over 98% of the urine samples metabolites of all five phthalates were detectable indicating a ubiquitous exposure of the German population to all five phthalates throughout the last 20 years. The median daily intakes in the subsets between 1988 and 1993 were quite constant for DnBP (approx. 7 microg/kg bw/d) and DEHP (approx. 4 microg/kg bw/d). However, from 1996 the median levels of both phthalates decreased continuously until 2003 (DnBP 1.9 microg/kg bw/d; DEHP 2.4 microg/kg bw/d). By contrast, the daily intake values for DiBP were slightly increasing over the whole time frame investigated (median 1988: 1.1 microg/kg bw/d; median 2003: 1.4 microg/kg bw/d), approximating the levels for DnBP and DEHP. For BBzP we observed slightly decreasing values, even though the medians as of 1998 levelled off at around 0.2 microg/kg bw/d. Regarding daily DiNP exposure we found continuously increasing values, with the lowest median being 0.20 microg/kg bw/d for the subset of 1988 and the highest median for 2003 being twice as high. The trends observed in phthalate exposure may be associated with a change in production and usage pattern. Female subjects exhibited significantly higher daily intakes for the dibutyl phthalates (DnBP p=0.013; DiBP p=0.004). Compared to data from US National Health and Nutrition Examination Surveys (NHANES) exposure levels of the dibutyl phthalates were generally higher in our German study population, while levels of BBzP were somewhat lower. Overall, for a considerable 14% of the subjects we observed daily DnBP intakes above the tolerable daily intake (TDI) value deduced by the European Food Safety Authority (EFSA) (10 microg/kg bw/d). However, the frequency of exceedance decreased during the years and was beneath 2% in the 2003 subset. Even though transgressions of the exposure limit values of the EFSA and the US Environmental Protection Agency (US EPA) occurred only in a relatively small share of the subjects, one has to take into account the cumulative exposure to all phthalates investigated and possible dose-additive endocrine effects of these phthalates. PMID- 17400025 TI - Neuronal glia interactions at the vertebrate neuromuscular junction. AB - Emerging studies demonstrate that perisynaptic Schwann cells (PSCs), which are the glia cells juxtaposed to the motor nerve terminal, actively participate in multiple aspects of the neuromuscular junction. During development, PSCs guide and promote synaptic growth. In adult muscles, PSCs can sense nerve stimulation by increasing intracellular calcium and are also capable of modulating transmitter release. Although adult PSCs are not required for acute synaptic maintenance and function, they are indispensable for long-term synaptic maintenance. Furthermore, PSC sprouts lead nerve terminal extension during synaptic remodeling. After nerve injury, PSCs sprout profusely and PSC processes guide regenerating nerve terminals. Future challenges will be to identify the molecular mechanisms by which PSCs interact with the nerve terminal and the muscle fiber. PMID- 17400026 TI - Prevention of birth asphyxia: responding appropriately to cardiotocograph (CTG) traces. AB - Birth asphyxia is a broad term that refers to intrapartum asphyxia sufficient to cause neurological damage in some newborns and, rarely, intrapartum or neonatal death. Cerebral palsy and long-term neurological complications such as learning difficulties and motor impairments may be due to causes other than birth asphyxia. Several intrapartum events may cause asphyxia (i.e. hypoxia and metabolic acidosis) leading to the likelihood of neurological injury. The cardiotocograph (CTG) is a screening tool that is used to assess fetal well-being during labour and to identify the possibility of asphyxia. Abnormality of the CTG, sometimes severe enough to be described as a pathological trace, is commonly termed 'fetal distress', although many fetuses with such traces may not have hypoxia and metabolic acidosis. In current practice, the events are appropriately termed 'pathological CTG trace' or 'acidotic pH' rather than 'fetal distress'. Accurate interpretation of CTG is essential, and it is important to recognize a fetus that shows a pathological CTG in labour that may imply possible hypoxia and birth asphyxia. Considering the wider clinical picture in interpreting the CTG, and taking timely and appropriate action based on the findings, may help prevent birth asphyxia. PMID- 17400027 TI - Sudden unexpected death in epilepsy: are winter temperatures a new potential risk factor? PMID- 17400028 TI - Correction of preoperative vitamin D deficiency after Roux-en-Y gastric bypass surgery. AB - BACKGROUND: To evaluate the adequacy of supplementation to correct preoperative vitamin D deficiency in adult patients during the year after Roux-en-Y gastric bypass (RYGB) surgery. METHODS: The medical records were reviewed and the preoperative and 12-month postoperative serum 25-hydroxyvitamin D [25(OH)D] levels were compared in patients who underwent RYGB from 2002 to 2004. The serum 25(OH)D levels were defined as being optimal (> or = 80 nmol/L), suboptimal (50 79 nmol/L), or deficient (<50 nmol/L). Patients with deficient 25(OH)D levels were prescribed 50,000 IU ergocalciferol weekly. The remaining patients averaged 710 IU supplemental vitamin D intake daily. RESULTS: The mean patient age was 43.8 +/- 10.7 years, and the mean preoperative body mass index was 51.8 +/- 9.8 kg/m2. Of the 95 patients with baseline and 12-month 25(OH)D levels, 89% were women. The mean preoperative 25(OH)D level was 49.7 +/- 26.5 nmol/L; 34% had suboptimal 25(OH)D levels and 54% had deficient levels before surgery. Twelve months after surgery, those receiving 50,000 IU weekly (n = 40) had a mean 25(OH)D level of 69.2 +/- 22.2 nmol/L; 63% had suboptimal and 8% deficient levels. Those taking 710 IU daily (n = 55) had a mean 25(OH)D level of 85.5 +/- 33.0 nmol/L; 44% had suboptimal and 6% deficient levels. CONCLUSION: Vitamin D deficiency is prevalent in RYGB patients before surgery. The vitamin D status improved markedly after RYGB surgery with either 710 IU vitamin D daily or 50,000 IU weekly. Current supplementation practices do not appear to optimize the serum 25(OH)D levels and need to be more closely examined. PMID- 17400029 TI - Comparison of laparoscopic linear staplers in clinical practice. AB - BACKGROUND: Only one previous study has evaluated the clinical safety of the 2 laparoscopic linear stapling devices available. Our study compared these staplers using laparoscopic gastric bypass as the standard procedure. METHODS: A total of 400 consecutive patients were prospectively studied. Group A (200 patients) underwent gastric bypass with the Endo-GIA Universal 6-row stapler (Autosuture) used, and group B (200 patients) underwent the procedure with the Ethicon 6-row stapler used. The measured parameters included stapler misfires, staple line bleeding, staple line leaks, unexplained gastrointestinal bleeding, unexplained intra-abdominal bleeding, total bleeding events, and total adverse events. RESULTS: Both groups were similar in preoperative body mass index, age, and gender. Both groups had a single staple line leak (0.5%; P = 1.0). Three (0.25% of staple firings) misfires occurred in group B. No misfires occurred in group A (P = .25). Gastrointestinal bleeding, staple line bleeding, and unexplained intra abdominal bleeding occurred in 6 (3%), 2 (1%), and 5 (2.5%) patients in group A and in 1 (0.5%), 0 (0%), and 2 (1%) patients in group B (P = .12, P = .5, and P = .45, respectively). Total bleeding events occurred in 13 patients (6.5%) in group A and in 3 patients (1.5%) in group B (P = 0.019). The total number of patients with an adverse event was 14 (7%) in group A and 7 (3.5%) in group B (P = .18). CONCLUSION: The results of our study have shown that the Ethicon 6-row stapler had more misfires and the Autosuture Endo-GIA 6-row stapler resulted in more bleeding complications. Only the difference in total bleeding events reached statistical significance but the difference in the total number of adverse events was not statistically significant. Therefore, in our experience both devices were equally safe and effective. PMID- 17400030 TI - Association between support group attendance and weight loss after Roux-en-Y gastric bypass. AB - BACKGROUND: Attending support group meetings has been linked to increased weight loss after gastric bypass surgery. However, the degree to which support group attendance influences weight loss is still unclear. This study quantitatively described the association between support group attendance and weight loss after Roux-en-Y gastric bypass. METHODS: The weight loss data and support group attendance of 78 consecutive Roux-en-Y gastric bypass patients were studied retrospectively. The patients were analyzed in 2 groups: those who attended >5 monthly support group meetings (group A) compared with those who went to < or =5 support group meetings (group B). The data from the first 12 months after surgery were analyzed. RESULTS: Group A achieved a mean percentage of excess weight loss of 10.5% at 2 weeks after surgery, 21.4% at 6 weeks, 30.9% at 3 months, 45.4% at 6 months, 53.6% at 9 months, and 55.5% at 12 months. Group B achieved a mean percentage of excess weight loss of 11.3% at 2 weeks, 21.8% at 6 weeks, 31.8% at 3 months, 41.3% at 6 months, 45.2% at 9 months, and 47.1% at 12 months. The differences between the 2 groups were significant at P <0.05 at 9 and 12 months. The weight loss was nonlinear and slowed as patients approached 1 year after surgery. CONCLUSION: Support groups are important for maintaining weight loss throughout the first year after surgery, especially after 6 months when the rate of weight loss from surgery begins to naturally decline. The amount of postoperative weight loss was greater than, or comparable to, the published data. Implementing regular support groups within the postoperative follow-up care may provide patients with the best chances of achieving maximal weight loss. PMID- 17400031 TI - Comparing outcomes of hand-assisted versus total laparoscopic gastric bypass. AB - BACKGROUND: Previous studies have compared open versus hand-assisted and open versus total laparoscopic Roux-en-Y gastric bypass. This study compared hand assisted and total laparoscopic Roux-en-Y gastric bypass surgery performed at 2 institutions by 2 surgeons, each specializing in 1 of the techniques. METHODS: Gastric bypass operations (n = 272) were evaluated from January 1, 2004 to December 31, 2004. The comparisons between the hand-assisted (n = 149) and laparoscopic (n = 123) approach were done using the median test, t test, chi square analysis, and Fisher's exact test. RESULTS: The average patient age was 43.9 years, and 84% (229 of 272) of the patients were women. The average body mass index was 48.8 kg/m2. No significant difference was found between the hand assist and laparoscopic groups in age, gender, preoperative body mass index, diabetes, hypertension, or operative time. No significant difference was found in the mortality rate between the hand-assist (2 of 149, 1.3%) and laparoscopic (1 of 123, 0.8%) groups. Also, no significant difference was found in the incidence of leaks, pulmonary embolism, deep venous thrombosis, wound infection, bowel obstruction, incisional hernia, fistula, persistent vomiting, or reoperation between the 2 procedures. A significant difference was found in the length of stay, with a median of 3 days for the hand-assist group and 2 days for the laparoscopic group (P <.001), and hemorrhage requiring transfusion, which occurred in 1 (.7%) of 149 hand-assist patients and 7 (5.7%) of 123 laparoscopic patients (P = .025). The percentage of excess weight loss 6 months after surgery was also similar between the two groups, at 50.2% and 52.1% for the hand-assist and laparoscopic groups, respectively (P = .45). CONCLUSIONS: The results of this study have found that both hand-assist and laparoscopic Roux-en-Y gastric bypass can be accomplished with similar perioperative risks and outcomes, including weight reduction. The choice between the 2 procedures depends on surgeon preference. PMID- 17400032 TI - Sleeve gastrectomy with wrapping using polytetrafluoroethylene to prevent gastric enlargement in a porcine model. AB - BACKGROUND: The safety and efficacy of laparoscopic sleeve gastrectomy followed by biliopancreatic diversion with duodenal switch for morbid obesity has been well established. We previously recommended 2-stage laparoscopic biliopancreatic diversion with duodenal switch for super-super obese patients. In the 2-stage version, these patients undergo laparoscopic sleeve gastrectomy as a first-stage procedure, followed by laparoscopic biliopancreatic diversion with duodenal switch as the second stage for more definitive treatment of their obesity. However, short-term weight regain may occur owing to gastric dilation after initial laparoscopic sleeve gastrectomy. The aim of this study was to prevent gastric dilation after sleeve gastrectomy. We designed a sleeve gastrectomy with wrapping using polytetrafluoroethylene dual mesh. METHODS: Eleven Yorkshire pigs weighing 20-25 kg underwent sleeve gastrectomy with wrapping using polytetrafluoroethylene dual mesh (wrapping group) or sleeve gastrectomy only (control group) to compare the weight loss. The animals were weighed weekly postoperatively. Necropsy was performed 8 weeks postoperatively to confirm the wrapping by pathologic report. RESULTS: Four pigs died because of staple line failure or strangulation; no perioperative complications occurred in the other pigs. The operative time for the control group was 198 +/- 60 minutes and for the wrapping group was 181 +/- 86 minutes (P = NS). The average weight of the removed stomach was 123.3 +/- 5.8 g in the control group and 140.3 +/- 69.9 g in the wrapping group (P = NS). The postoperative weight gain at 8 weeks was significantly slower in the wrapping group than in the control group (P <.0001). The pathologic necropsy report noted that the mesh was well attached to the stomach wall at 9 days postoperatively, with no unexpected deaths. CONCLUSION: Sleeve gastrectomy with wrapping using polytetrafluoroethylene dual mesh is feasible, and weight gain was reduced in the porcine model. PMID- 17400033 TI - Band versus bypass: influence of an educational seminar and surgeon visit on patient preference. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass and laparoscopic adjustable gastric banding are 2 common weight loss procedures. This investigation examined the effect of a preoperative educational seminar (ES) and surgeon visit (SV) on patients' choice of bariatric procedure. METHODS: In our practice, patients choose their procedure. New patients receive an overview of both procedures in an ES, including the risks and benefits, and then meet individually with a surgeon (SV) to answer any additional questions. Three identical surveys (before the ES, after the ES, and after the SV) were given to new patients who voluntarily participated in this study. The survey queried procedure choice and influencing factors. RESULTS: A total of 47 patients participated. Of these 47 patients, 31 had researched the procedures before the ES and 13 were unsure of the differences between laparoscopic Roux-en-Y gastric bypass and laparoscopic adjustable gastric banding before the ES, 4 were after the ES, and 1 was after the SV. Also, 11% of patients changed their procedure choice as a result of attending the ES and SV; 15%, 13%, and 13% of patients were willing to be randomized to either procedure before the ES, after the ES, and after the SV, respectively. CONCLUSIONS: Only 11% of patients changed their procedure choice as a result of attending the ES and SV. Thus, patient decisions are usually made before meeting the surgeon, and the information provided at the ES and SV simply reinforced those decisions. Only 13% of patients were willing (4% very willing and 9% somewhat willing) to be randomized to either procedure (laparoscopic Roux-en-Y gastric bypass or laparoscopic adjustable gastric banding). It is likely that patients had confidence in, and were comfortable with, 1 procedure over the other, and therefore were unwilling to undergo the other procedure. PMID- 17400034 TI - Avoiding and resolving conflicts between DNA replication and transcription. AB - DNA replication and transcription are essential processes for cell growth and division. The polymerase complexes charged with these tasks share the same template and so there is a potential for conflict between the two. Studies both in vivo and in vitro indicate that RNA polymerase complexes can indeed arrest the progress of replication forks and that cell viability is endangered as a result. Stable ternary complexes formed at lesions in the template DNA or as a result of backtracking may be especially troublesome. Numerous processes have evolved to reduce conflict between replication and transcription and to rescue any replication forks that suffer damage when conflict does occur. In bacteria, these systems are so highly tuned and co-ordinated that cells normally suffer few problems even under conditions allowing rapid growth and therefore demanding both efficient chromosome duplication and high levels of gene expression. The threat to genome transmission and cell viability becomes apparent when this interplay is disrupted. PMID- 17400035 TI - The influence of body composition on respiratory muscle, lung function and diaphragm thickness in adults with cystic fibrosis. AB - BACKGROUND: Weight loss and loss of fat-free mass (FFM) are associated with peripheral muscle wasting in cystic fibrosis (CF) although whether this co-exists with loss of diaphragm mass remains unclear. METHODS: FFM was determined by dual energy X-ray absorptiometry and bioelectrical impedance in 40 adults with CF and 30 age-matched healthy subjects (HS). Diaphragm thickness at functional residual capacity (FRC) [TDIrel] and total lung capacity (TLC) [TDIcont] and thickening ratio (TR) were assessed by ultrasonography. Inspiratory muscle strength and work capacity were determined by maximal inspiratory pressure (PImax), and sustained PImax (SPImax); pulmonary function (RV, VC and TLC) and physical activity status (PAS) were also determined. RESULTS: When the CF patients were assessed as a group (low and normal FFM) they had similar age, weight, height and PAS compared to the HS, although patients had lower FFM (p<0.05), VC and TLC than the HS (p<0.01). In addition, although PImax, TDIrel, TDIcont and TR were similar between the patients and the HS, SPImax was lower in the patients (p<0.01). When analyses were made between patients with low versus normal FFM and between patients with low FFM and HS no significant differences were found between overall weight although TDIrel, TDIcont, TR and PAS were all reduced in patients with low FFM (p<0.01). CONCLUSIONS: PImax is relatively well preserved in adults with CF although there is a relationship between the loss of inspiratory muscle work capacity, FFM, PAS and pulmonary function. Furthermore loss of FFM is associated with loss of diaphragm muscle mass. PMID- 17400036 TI - Metabolic profiling of a potential antifungal drug, 3-(4-bromophenyl)-5 acetoxymethyl-2,5-dihydrofuran-2-one, in mouse urine using high-performance liquid chromatography with UV photodiode-array and mass spectrometric detection. AB - 3-(4-bromophenyl)-5-acetyloxymethyl-2,5-dihydrofuran-2-one (LNO-18-22) is a representative member of a novel group of potential antifungal drugs, derived from a natural 3,5-disubstituted butenolide, (-)incrustoporine, as a lead structure. This lipophilic compound is characterized by high in vitro antifungal activity and low acute toxicity. For the purpose of in vivo studies, a new bioanalytical high-performance liquid chromatographic method with UV photodiode array and mass spectrometric detection (HPLC-PDA-MS), involving a direct injection of diluted mouse urine was developed and used in the evaluation of the metabolic profiling of this drug candidate. The separation of LNO-18-22 and its phase I metabolites was performed in 37 min on a 125 mmx4 mm chromatographic column with Purospher RP-18e using an acetonitrile-water gradient elution. Scan mode of UV detection (195-380 nm) was employed for the identification of the parent compound and its biotransformation products in the biomatrix. Finally, the identity of LNO-18-22 and its metabolites was confirmed using HPLC-MS analyses of the eluate. These experiments demonstrated the power of a comprehensive analytical approach based on the combination of xenobiochemical methods and the results from tandem HPLC-PDA-MS (chromatographic behaviour, UV and MS spectra of native metabolites versus synthetic standards). The chemical structures of five phase I LNO-18-22 metabolites and one phase II metabolite were elucidated in the mouse urine, with two of these metabolites having very unexpected structures. PMID- 17400037 TI - Simultaneous determination of 102 pesticide residues in Chinese teas by gas chromatography-mass spectrometry. AB - An efficient and sensitive method for simultaneous determination of 102 pesticide residues in teas has been established and validated. The multi-residue analysis of the pesticides in teas involved extraction with acetone-ethyl acetate-hexane, clean-up using gel permeation chromatography (GPC) and solid-phase extraction (SPE), and subsequent identification and quantification of the selected pesticides by gas chromatography-mass spectrometry (GC-MS) under retention time locked (RTL) conditions. For most of the target analytes, the optimized pretreatment processes led to no significant interference on analysis from sample matrix, and the determination of 102 compounds was achieved in about 120 min. Pesticide residues could be determined in low sub-ppb range, from 0.01 microg/mL for hexachlorobenzene to 2.5 microg/mL for propargite, with average recoveries ranging from 59.7 to 120.9% (mean 88%) and relative standard deviations (RSDs) in the range 3.0-20.8% (mean 13.7%) for all analytes across three fortification tea levels. The limits of detection (LODs) were much lower than the maximum residue levels established by the European Union (EU) legislations. PMID- 17400038 TI - Studies on preparing and adsorption property of grafting terpolymer microbeads of PEI-GMA/AM/MBA for bilirubin. AB - Crosslinking copolymer microbeads with a diameter range of 100-150 microm were synthesized by suspension copolymerization of glycidyl methacrylate (GMA), acrylamide (AM) and N,N'-methylene bisacrylamide (MBA). Subsequently, polyethyleneimine (PEI) was grafted on the surfaces of the terpolymer microbeads GMA/AM/MBA via the ring-opening reaction of the epoxy groups, and the grafting microbeads PEI-GMA/AM/MBA were prepared. In this paper, the adsorption property of the grafting microbeads for bilirubin was mainly investigated, and the effects of various factors, such as pH value, ionic strength and grafting degree of PEI on the surface of grafting microbeads and the adsorption capacity of the grafting microbeads for bilirubin were examined. The batch adsorption experiment results show that by right of the action of grafted polyamine macromolecules PEI, the grafting microbeads PEI-GMA/AM/MBA have quite strong adsorption ability for bilirubin; the isotherm adsorption conforms to Freundlich equation. The pH value of the medium affects the adsorption capacity greatly, As in the nearly neutral solutions with pH 6, the grafting microbeads have the strongest adsorption ability for bilirubin, whereas in acidic and basic solutions their adsorption ability is weak. The ionic strength hardly affects the adsorption ability of the grafting microbeads. The grafting degree of PEI on the surfaces of the grafting microbeads also has a great effect on the adsorption capacity, and higher the grafting degree of PEI on the surface of the microbead PEI-GMA/AM/MBA, the stronger is the adsorption ability of the microbeads. PMID- 17400039 TI - Accurate quantification of dimethylamine (DMA) in human plasma and serum by GC-MS and GC-tandem MS as pentafluorobenzamide derivative in the positive-ion chemical ionization mode. AB - Dimethylamine (DMA) circulates in human blood and is excreted in the urine. Major precursor for endogenous DMA is asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide (NO) synthesis. ADMA is hydrolyzed to DMA and L citrulline by dimethylarginine dimethylaminohydrolase (DDAH). In previous work, we reported a GC-MS method for the quantification of DMA in human urine. This method involves simultaneous derivatization of endogenous DMA and the internal standard (CD(3))(2)NH by pentafluorobenzoyl chloride (PFBoylCl) and extraction of the pentafluorobenzamide derivatives by toluene. In the present work, we optimized this derivatization/extraction procedure for the quantitative determination of DMA in human plasma. Optimized experimental parameters included vortex time and concentration of PFBoylCl, carbonate and internal standard. The GC-MS method was thoroughly validated and applied to measure DMA concentrations in human plasma and serum samples. GC-MS quantification was performed by selected ion monitoring of the protonated molecules at m/z 240 for DMA and m/z 246 for (CD(3))(2)NH in the positive-ion chemical ionization mode. Circulating DMA concentration in healthy young women (n=18) was determined to be 1.43+/-0.23 micaroM in serum, 1.73+/-0.17 microM in lithium heparin plasma, and 9.84+/-1.43 microM in EDTA plasma. DMA was identified as an abundant contaminant in EDTA vacutainer tubes (9.3+/-1.9 nmol/monovette, n=6). Serum and lithium heparin vacutainer tubes contained considerably smaller amounts of DMA (0.42+/-0.01 and 0.95+/-0.01 nmol/monovette, respectively, each n=6). Serum is recommended as the most appropriate matrix for measuring DMA in human blood. The present GC-MS method should be useful for the determination of systemic and whole body DDAH activity by measuring circulating and excretory DMA in experimental and clinical studies. PMID- 17400040 TI - Val326 of Thermoactinomyces vulgaris R-47 amylase II modulates the preference for alpha-(1,4)- and alpha-(1,6)-glycosidic linkages. AB - Thermoactinomyces vulgaris R-47 alpha-amylase II (TVA II) catalyzes not only the hydrolysis of alpha-(1,4)- and alpha-(1,6)-glycosidic linkages but also transglycosylation. The subsite +1 structure of alpha-amylase family enzymes plays important roles in substrate specificity and transglycosylation activity. We focused on the amino acid residue at the 326th position based on information on the primary structure and crystal structure, and replaced Val with Ala, Ile, or Thr. The V326A mutant favored hydrolysis of the alpha-(1,4)-glycosidic linkage compared to the wild-type enzyme. In contrast, the V326I mutant favored hydrolysis of the alpha-(1,6)-glycosidic linkage and exhibited low transglycosylation activity. In the case of the V326T mutant, the hydrolytic activity was almost identical to that of the wild-type TVA II, and the transglycosylation activity was poor. These results suggest that the volume and the hydrophobicity of the amino acid residue at the 326th position modulate both the preference for glycosidic linkages and the transglycosylation activity. PMID- 17400041 TI - Treatment experience in a child with heterochronous bilateral blowout fractures. PMID- 17400042 TI - Harvesting a dorsal digital vein and its intercapitular branch at the MCPJ level in digital replantation. PMID- 17400043 TI - Skeletal effects of long-term estrogen and testosterone replacement treatment in a man with congenital aromatase deficiency: evidences of a priming effect of estrogen for sex steroids action on bone. AB - The relative contribution of each sex steroid (i.e. estrogen and androgen) on bone in men and the relationships among sex steroids and changes in BMD and bone strength are still unknown. A defective BMD of bone tissue is constantly present in men with aromatase deficiency. This study evaluates the effects of different regimens of treatment with sex steroids over 7.3 years follow-up on BMD in an adult man affected by aromatase deficiency and by a concomitant mild hypogonadism, as previously described. The aim of the study is to provide additional data on the relative roles of androgens and estrogens in male bone metabolism. The effects of testosterone (T) treatment alone and estrogen (tE(2)) treatment alone as well as the effects of the combined treatment with testosterone and estradiol (T plus tE(2)) on areal BMD (aBMD) at dual-energy X ray absorptiometry (DXA) and the effects of T plus tE(2) on volumetric BMD (vBMD), particular at cortical site, measured by peripheral quantitative computed tomography (pQCT), are investigated. Hormones and markers of bone turnover were monitored during all phases of the study. Treatment with tE(2) normalized serum estradiol, but only the combined treatment with T plus tE(2) normalized both serum estradiol and testosterone. Markers of bone turnover reached a pattern close to normality during T plus tE(2). The aBMD was little modified by T, but increased more during tE(2). T plus tE(2) resulted in a further increase in both aBMD at DXA and vBMD at pQCT. Cortical thickness increased during T plus tE(2) both in radius and tibia. Only the combined treatment led to optimal parameters of aBMD suggesting that testosterone needs estrogens as a permissive factor for a direct androgen anabolic action on bone in men. PMID- 17400044 TI - Allometric scaling and biomechanical behavior of the bone tissue: an experimental intraspecific investigation. AB - INTRODUCTION: Adaptation of bone to different loads has received much attention. This paper examines the consequences of differences in size on bones from the same animal species. METHODS: The study was conducted on 32 canine radii. Their geometry, densitometry and mechanical properties were determined and one-way ANOVA was used to analyze their distribution by sex. Bending failure was observed during the mechanical test. The bones were then likened to thin beams and the mechanical parameters of interest were appraised via beam theory. A multiple linear regression model with stepwise analyses was employed to determine which parameters rule the mechanical characteristics. The relationships between the bone mass and the parameters investigated were analyzed by means of a model II regression in order to state how the scaling of the bone characteristics act on its mechanical behavior. RESULTS: The linear regression model demonstrated that an animal's mass, its sex and the mineral content and the geometrical properties of its bones almost entirely predict their mechanical behavior. A close fit was found between the experimentally determined and the theoretical slopes of the log regressed allometric equations. The work to failure was found to scale almost linearly with the animal and bone mass and the macroscopical bone material properties were found to be mass invariant. The allometric equations showed that as the animal mass increases, employing proportionally the same amount of tissue, bones get proportionally shorter and proportionally distribute their tissue further from the cross-sectional centroid. CONCLUSIONS: Our results suggest that dimensional analysis on the assumption of geometrical self-similarity and mechanical testing according to classic elastic solutions are reasonable in bones tested in accordance to our set up. The bone geometry is the parameter able to curb the energy effects of an animal mass increase. The allometric scaling of the bone length and the cross-sectional layout, without an increase in the amount of material proportionally employed, preserves linear with the animal mass the amount of energy necessary to fracture a bone and restrain the rise of stresses and strains in the cross-section. PMID- 17400046 TI - The role of maternal affect attunement in dyadic and triadic communication. AB - The influence of maternal affect attunement on the relationship between gaze monitoring during dyadic communication at 3 months and coordinated attention during triadic communication at 5, 7 and 10 months was examined in a longitudinal study. Although most infants engaged in gaze monitoring at 3 months and in coordinated attention at 5, 7 and 10 months, a regression analysis revealed that gaze monitoring at 3 months significantly predicted coordinated attention at 10 months only when maternal affect attunement was high. These findings are discussed in terms of theories that emphasize the role of social interaction in the development of meaningful communication and continuity in mental state awareness during the first year of life. PMID- 17400047 TI - Toy-oriented changes in early arm movements II--joint kinematics. AB - Our recent work suggests that infants begin to change their hand and joint kinematics in the presence of a toy months before the onset of purposeful reaching. Moreover, these 'toy-oriented' changes in hand kinematics cluster into Early, Mid and Late phases. The purpose of the present study was to test hypotheses regarding toy-oriented changes in joint kinematics in the same infants. METHODS: Thirteen infants were observed every other week from 8 weeks up to the first week of reaching around 20 weeks. At each session, a high-speed motion analysis system recorded infants' arm movements with and without a toy present. RESULTS: During the Early phase, infants scaled down their movements. In contrast, during the Mid phase infants scaled up their movements and did not change the relationship between the shoulder and elbow for speed and smoothness related variables. In addition, infants showed toy-oriented changes such as increase in shoulder flexion and adduction. In the Late phase, infants continued to produce toy-oriented changes in shoulder orientation, and increased shoulder excursion and speed relative to the elbow. Thus, the toy-oriented changes in hand kinematics in the Mid and Late phases [Bhat, A. N., & Galloway, J. C. (2006). Toy oriented changes in early arm movements of young infants: Hand kinematics. Infant Behavior and Development, 29(3), 358-372] more closely followed changes in shoulder kinematics. Lastly, results are discussed in terms of shoulder-elbow dissociations, speed-amplitude relationships, and the key role of spontaneous movements in the development of reaching. PMID- 17400048 TI - Imitating step by step: a detailed analysis of 9- to 15-month-olds' reproduction of a three-step action sequence. AB - To investigate what infants around their first birthday learn from observing an action sequence, 9- to 15-month-olds' imitative behavior was compared in three conditions: a demonstration group watched three target action steps and a final outcome, a control group observed only the third step and final outcome, and a baseline group received no demonstration. After a short delay, the demonstration infants of all ages produced more target actions than the control and baseline infants. Moreover, the latency to the first step was shortest in the demonstration condition. Data about the performance of the single steps in each age sample revealed which target actions were novel to the infants, and which steps were learned by observation. Infants in the control condition did not generate the unseen target action steps. It is concluded that infants' memory capacity and their ability to encode action-effect relations contribute to their imitation of an action sequence. PMID- 17400050 TI - Reformulating infant and toddler social competence with peers. AB - Efforts to identify children at risk for social and academic difficulties at an early age have begun to apply conceptualizations of social competence with peers from childhood to infancy and toddlerhood. These attempts have had limited success, in part because social behaviors studied in later childhood such as negative or aggressive acts may not capture relevant dimensions of social competence with peers during the infant and toddler years. The present study conducted an exploratory factor analysis to begin developing a conceptualization of social competence with peers that is appropriate for use with infants and toddlers, and that captures individual differences capable of predicting later social functioning. Results indicate three dimensions of infant and toddler social competence with peers: peer sociability, active peer refusal, and passive peer avoidance. Negative and aggressive behaviors loaded on each of the three factors rather than emerging as a unique dimension of social competence. Each factor demonstrated both convergent and predictive validity, indicating that early identification of social skills remains a reasonable goal for practitioners and researchers alike. Finally, antecedents within children, families, and out-of home child care environments helped to explain individual variability in each of these three factors of social competence with peers. PMID- 17400049 TI - Maternal sensory sensitivity, mother-infant 9-month interaction, infant attachment status: predictors of mother-toddler interaction at 24 months. AB - At 24-months of age, toddlers (N=62) and their mothers were observed in a free play session to determine the contribution of (a) maternal sensory sensitivity to positive and negative infant facial expressions as measured in a signal detection task at 6 months, (b) maternal behavior and affect, infant behavior and affect, and dyadic interaction at 9 months, and (c) infant attachment status at 12 months in predicting maternal, toddler, and dyadic measures at 24 months. Hierarchical regression analyses revealed that over and above early maternal behavior, which was predictive of later maternal behavior at 24 months, sensory sensitivity to the positive infant expression at 6 months predicted maternal behavior at 24 months and sensory sensitivity to both the positive and negative expression was associated with later maternal affect. Infant attachment status emerged as the variable which predicted toddler behavior and dyadic interaction at 24 months. PMID- 17400051 TI - Joint visual attention and social triangular engagement at 3 and 6 months. AB - Two experiments document infants' ability to detect breaks of contact in an ongoing dyadic interaction and to use the joint attention mechanism for social triangular communication purposes. In Experiment 1, the ability of 3- and 6-month olds to show joint attention when an adult turned to face another person in sight, and to react when both adults conversed in a reciprocal manner, was measured. In Experiment 2, 3-month-olds were tested in two joint attention conditions: a Person-Person-Object condition (PPO) and a Person-Person-Person condition (PPP). Results showed that both 3- and 6-month-olds' gazed backwards and forwards between the two adults and produced socially directed behaviors towards the adults, thus initiating a social triangular coordinated interaction. Three-month-olds replicated this pattern in the PPP condition but not in the PPO condition. The social context with three persons may be particularly facilitative for infant early communication and sharing within the family. PMID- 17400052 TI - Influence of vocal cues on learning about objects in joint attention contexts. AB - An experimenter taught infants about a novel toy in two joint attention conditions, one with and one without vocal cues. In test trials, infants viewed the familiar toy and a novel toy. Infants in the Joint Attention plus Voice condition looked significantly longer to the novel toy. PMID- 17400053 TI - 8th international congress of neuroimmunology. PMID- 17400054 TI - Proceedings from the 4th International Conference on HLA-G, July 2006, Paris, France. PMID- 17400055 TI - Structural studies on HLA-G: implications for ligand and receptor binding. AB - Human leukocyte antigen-G (HLA-G) is a class Ib major histocompatibility complex (MHC) molecule that is specifically expressed in immune-privileged tissues. The overall structure of HLA-G resembles other class I MHC molecules, in which a heavy chain comprised of three domains is noncovalently associated with beta(2)microglobulin (beta(2)m). A nine-residue self-peptide is bound within a cleft formed by two alpha-helices and a beta-sheet floor. An extensive network of contacts is formed between the peptide and the binding cleft, leading to a constrained mode of binding reminiscent of that observed in HLA-E. The alpha3 domain of HLA-G, the putative binding site for leukocyte immunoglobulinlike receptor-1 (LIR-1) and -2, is structurally distinct from class Ia MHC molecules, providing a basis for the observed differences in affinity for these ligands. In addition, a disulfide-bonded dimer adopts an oblique conformation, providing the possibility of a 1:2 (HLA-G dimer:receptor) complex stoichiometry. The relative orientation of the HLA-G protomers in the dimer structure suggests that it is unlikely that dimerization is involved in killer immunoglobulinlike receptor 2DL4 (KIR2DL4) binding. PMID- 17400056 TI - HLA-G complexes are observed on the cell surface. AB - In the context of pregnancy, several immunomodulating mechanisms have developed to regulate the maternal immune response to its semiallogeneic fetus. The nonclassical major histocompatibility complex class I human leukocyte antigen-G (HLA-G) was suggested to be involved in these mechanisms due to its unique features and its immunosuppressive abilities. We have previously described the presence of HLA-G complexes at the cell surface, which confer an efficient natural killer inhibition through the leukocyte Ig-like receptor-1 (LIR-1). We further demonstrated the presence of HLA-G free heavy chain (FHC) complexes, which are not recognized and possibly interfere with LIR-1 and HLA-G interaction. Here we expand our understanding of the nature of the complexes by demonstrating that these complexes are observed mainly on the cell surface and not inside the cell. We further determine that the HLA-G stability at the cell surface is not a direct result of the presence of the HLA-G complexes. Finally, we suggest that the FHC complexes are probably assembled from the conformed complexes present on the cell surface. PMID- 17400057 TI - Soluble HLA-G and HLA-G1 expressing antigen-presenting cells inhibit T-cell alloproliferation through ILT-2/ILT-4/FasL-mediated pathways. AB - HLA-G is a tolerogenic molecule involved in maternal-fetal tolerance and in allograft acceptance. Soluble HLA-G proteins are present at high levels in plasma from transplanted patients who better accept their graft. In addition, infiltrating mononuclear cells expressing HLA-G can be detected within grafted tissues. To define the role of these HLA-G proteins in preventing graft rejection, we investigated the ability of HLA-G1 expressing antigen presenting cells (APC) and of soluble HLA-G proteins (i.e., HLA-G5 and shed HLA-G1) to inhibit T-cell alloproliferation and analyzed the molecules involved in such inhibition. Results demonstrated that both membrane-bound and soluble HLA-G proteins inhibited T-cell alloproliferation. This inhibition involved engagement of immunoglobulinlike transcript (ILT)-2 and ILT-4 receptors by HLA-G. Moreover, blocking Fas ligand (FasL) reversed HLA-G mediated inhibition, demonstrating that the Fas/FasL pathway is also recruited by HLA-G to exert its immunosuppressive function on T cells. These data highlight the role played by HLA-G in better graft acceptance status observed in transplanted patients with HLA-G(+) grafted cells and high HLA-G plasma levels. Evidence to support such role in vivo was provided by the capacity of purified HLA-G5 from the plasma of the transplanted patient to suppress T-cell alloresponses. PMID- 17400058 TI - Exchanges of membrane patches (trogocytosis) split theoretical and actual functions of immune cells. AB - Exchanges of antigens between immune cells have long been evidenced in the murine system and more recently in humans, but the mechanisms by which these transfers occur, and even more so their functional and physiologic significance remain unclear. Yet, intercellular antigen exchanges, and particularly intercellular exchanges of intact membrane patches, also called trogocytosis, have recently been the subject of renewed interest. Indeed, trogocytosis has been thoroughly investigated in terms of phenomenology, mechanisms and parameters, and function. For lack of a dramatic function for trogocytosis, the possible significance of membrane patch transfers has been discussed. Here, we will briefly outline the key findings concerning trogocytosis, highlight their significance, and discuss how they have an impact on commonly accepted immune mechanisms. PMID- 17400059 TI - HLA-G expression is a fundamental prerequisite to pregnancy. AB - Human leukocyte antigen-G (HLA-G) is thought to play a key role in implantation by controlling trophoblast invasion and maintaining a local immunosuppressive state. The secretion of soluble HLA-G antigens (sHLA-G) by early embryos seems necessary for a successful implantation and could be a marker of increased pregnancy rate following in vitro fertilization. We have reviewed the results obtained during the last years (from 1987 to 2005). They overall confirmed the predictive role of sHLA-G production in pregnancy outcome. Furthermore, we have examined the technical procedures utilized, with a particular attention to the monoclonal antibodies used in the enzyme-linked immunosorbent assay (ELISA) techniques. New functional roles for HLA-G molecules in pregnancy could be suggested by the relationship observed between the presence of sHLA-G antigens in follicular fluids and sHLA-G expression in the corresponding fertilized oocyte. Furthermore, since maternal mRNA is fundamental for protein production in early embryos, the biologic role of the HLA-G 14 base pair polymorphism could be explored. PMID- 17400060 TI - Rapid evaluation of soluble HLA-G levels in supernatants of in vitro fertilized embryos. AB - Human leukocyte antigen G (HLA-G) molecules are crucial for the maternal tolerance against the fetus during pregnancy. Thus, the presence of soluble HLA-G (sHLA-G) in embryo cultures is thought to be correlated to a successful pregnancy after assisted reproductive techniques (ART). Here, we established a rapid detection assay based on Luminex technology, which can be integrated into ART proceedings, allowing sHLA-G quantification in sample volumes of only 10 microl within 1.5 hours. Using this method, sHLA-G levels of 526 single-embryo cultures, 47 two-embryo cultures, and 15 three-embryo cultures were analyzed corresponding to 313 ART cycles. In 117 embryo cultures, sHLA-G was detectable. In single embryo cultures, the sHLA-G levels were positively correlated to embryo quality (p = 0.048, r = 0.20, n = 100). The presence of sHLA-G in embryo cultures was significantly (p < 0.0001) associated with clinical pregnancy after intracytoplasmatic sperm injections (ICSI), especially in couples with male factor infertility, but not after in vitro fertilization (IVF) or in couples with female infertility. Importantly, in sHLA-G negative embryos, the abortion rate was increased threefold (p = 0.04). In conclusion, the results obtained by our novel method support strongly the diagnostic relevance of sHLA-G for predicting pregnancy outcome after ART. The ultimate conditions for this prediction have to be further investigated in a multicenter study. PMID- 17400061 TI - Implication of HLA-G in human embryo implantation. AB - The mechanisms underlying the processes of human embryo implantation and maternal immunotolerance to the fetus remain incompletely understood. Growing evidence indicates that one of the nonclassic human leukocyte antigen (HLA) genes, HLA-G, is expressed selectively on the surface of the extravillous throphoblast and plays an important role in the adaptations of the maternal immune system to pregnancy; however the implication of this molecule in the process of human embryo implantation is also plausible. Therefore the aim of the present article is to review the available literature specifically focused on the possible relationship between HLA-G and human embryo implantation. In particular, studies investigating HLA-G expression on human preimplantation embryos and in the endometrium, as well as its levels in embryo culture supernatants and circulating maternal blood, are discussed. PMID- 17400062 TI - Mechanisms of prolongation of allograft survival by HLA-G/ILT4-modified dendritic cells. AB - Engagement of inhibitory receptors on dendritic cells (DCs) is a powerful way to modulate their functions to achieve hyporesponsiveness or tolerance induction. Transgenic mice expressing human ILT4 receptor exclusively on DCs and triggered by HLA-G1 developed long-term survival of allogeneic skin transplant. Here we identify the cellular and molecular mechanisms responsible for that induction of hyporesponsiveness to alloantigen in vivo. Engagement of ILT4 receptor by HLA-G1 resulted in down-regulation of expression of MHC class II and costimulatory molecules, and modulation of cytokine production on DCs. HLA-G-modified DCs from ILT4 transgenic mice promote long-term survival of allografts by mechanisms involving both the induction of regulatory T cells and T-cell anergy. A novel feature of our research was to establish a model for the study of the prospective mechanisms of regulation of alloimmune responses by inhibitory receptors in vivo and analysis of the potential of HLA-G and its inhibitory receptors in modulation of DCs and T-cell function. PMID- 17400063 TI - Application of human leukocyte antigen-G expression in the diagnosis of human cancer. AB - It has been well known that human leukocyte antigen (HLA)-G molecules are present in a variety of human neoplastic diseases, and the molecule may contribute to the escape of tumor cells from immune surveillance. Besides the studies that aim at elucidating the roles of HLA-G in immune regulation, the researches that focus on potential applications of HLA-G expression in cancer diagnosis represent another perspective in HLA-G research. This review summarizes those recent translational studies of HLA-G expression in the diagnosis of human cancer. Specifically, the promises and challenges for applying HLA-G expression to detect cancer in body fluids, to diagnose different types of human cancer and to predict clinical outcome in cancer patients will be briefly reviewed. PMID- 17400064 TI - Hypoxia modulates HLA-G gene expression in tumor cells. AB - Human leukocyte antigen G (HLA-G) molecules are expressed in cytotrophoblasts and play a key role in maintaining immune tolerance at the maternal-fetal interface. HLA-G expression was also reported in inflammatory diseases, organ transplantation, and malignant tumors. The regulatory mechanisms of HLA-G gene expression differ from those of classical HLA class I genes and are still only partially elucidated. Focusing on tumor cells, we previously demonstrated a tight control of HLA-G gene expression by cis-acting epigenetic mechanisms. In the present study, we hypothesized that these processes are dependent of microenvironment conditions, and more particularly, stress conditions like hypoxia. Cellular response to hypoxia is mainly driven by a key transcription factor, hypoxia-inducible factor 1 (HIF-1), and other factors, such as NF-kappaB, involved in angiogenesis and cell survival. Here we confirmed the influence of hypoxia on HLA-G gene induction in the HLA-G-negative M8 melanoma cell line. Moreover, upon treatment with the hypoxia-mimicking desferrioxamine, we demonstrated a decrease in HLA-G gene expression in melanoma FON and choriocarcinoma JEG-3 cell lines, both expressing constitutively HLA-G. Finally, we demonstrated for the first time that the modulation of HLA-G gene expression is dependent of HIF-1 stabilization and thus might be relevant for the control of HLA-G gene expression in hypoxic tumors. PMID- 17400065 TI - HLA-G in the nervous system. AB - This review summarizes the current knowledge on the significance of human leukocyte antigen G (HLA-G) in the nervous system under physiologic and pathologic conditions. The central nervous system (CNS) has classically been viewed as an immune-privileged organ. Immune reactions triggered by, or directed against, CNS structures proceed along specific pathways. The expression and the functioning of the immune-tolerogenic major histocompatibility complex (MHC) molecule HLA-G have revealed novel insights into the endogenous immune-regulatory mechanisms exerted by resident cells within the nervous system, as well as how migrating immune cells contribute to this under pathologic conditions. HLA-G has been evidenced in certain neurologic disorders, including those of autoimmune, infectious, and neoplastic origin. This review compiles the current state of knowledge: how HLA-G is considered to be operative at different levels of the respective pathogenetic cascades of neurologic disorders. These findings are relevant both from a pathogenetic as well as from a therapeutic viewpoint. PMID- 17400066 TI - Modulation of HLA-G and HLA-E expression in human neuronal cells after rabies virus or herpes virus simplex type 1 infections. AB - Human Leukocyte Antigen (HLA)-G and E are nonclassical human MHC class I molecules. They may promote tolerance leading to virus and tumor immune escape. We recently described that the herpes simplex virus type 1 (HSV-1), a neurotropic virus inducing chronic infection and neuron latency, and rabies virus (RABV), a neuronotropic virus triggering acute neuron infection, up-regulate HLA-G expression in human neurons (NT2-N). Surface expression was only detected after RABV infection. We investigated here whether RABV and HSV-1 up-regulate HLA-E expression in human neuronal precursors (Ntera-2D/1). We found that RABV, not HSV 1, up-regulates HLA-E expression, nevertheless HLA-E could not be detected on the surface of RABV-infected Ntera-2D/1. Altogether these data suggest that HLA-G and not HLA-E could contribute to the immune escape of RABV. In contrast, there was no evidence that these molecules are used by latent HSV-1 infection. Thus, neurotropic viruses that escape the host immune response totally (RABV) or partially (HSV-1) regulate HLA-G expression on human neuronal cells differentially. PMID- 17400067 TI - Nucleoside reverse transcriptase inhibitors are able and protease inhibitors unable to induce the tolerogenic molecule HLA-G1 on monocytes from HIV-1 infected patients. AB - Our group has previously reported that a significantly larger proportion of peripheral monocytes from human immunodeficiency virus type 1 (HIV-1) seropositive individuals receiving highly active antiretroviral therapy (HAART) express HLA-G1 and also that one of the HAART components, the nucleoside reverse transcriptase inhibitors (NRTIs), may be involved in this effect. Because protease inhibitors (PIs) are another component of HAART that are administered with NRTIs, the aim of this work was to determine whether or not PIs are also involved in the HLA-G1 changes previously observed in treated HIV-1 positive patients. CD14(+) cells expressing HLA-G1 were therefore measured in 7 HIV-1 positive patients whose initial HAART was changed to a protease inhibitor-only regime due to drug toxicity and/or virologic resistance. Our results indicate that PIs do not appear to be implicated in the rise of HLA-G1 expression on CD14(+) cells from HIV-1 infected individuals receiving HAART, while we further confirm that NRTIs are involved in the surface induction of HLA-G1. PMID- 17400070 TI - Soft tissue wounds and principles of healing. AB - Wound healing is a complex interchange, orchestrated between cellular components that play their respective parts signaled by and mediated by different cellular instruments of healing. When healing is performed well, the final product is a thing of beauty. When healing is delayed, interrupted, or excessive, then unsightly scars of chronic painful wounds that are frustrating to the patient and physician occur. PMID- 17400071 TI - Principles of basic wound evaluation and management in the emergency department. AB - The primary objectives of basic wound management center around promoting optimal wound healing and cosmesis. These objectives may be achieved through the systematic assessment, preparation, and repair of the laceration supplemented with appropriate patient care instructions. The meticulous and methodical management of traumatic wounds described in this article will assist the emergency physician in decreasing overall complication rates and help improve patient satisfaction. PMID- 17400072 TI - Anesthetic and procedural sedation techniques for wound management. AB - Emergency physicians come across a wide variety of painful conditions and perform a large number of uncomfortable procedures on a typical shift. This article describes the local anesthetic agents and their potential applications. The peripheral nerve blocks that are regularly done in the emergency department are described. Lastly, procedural sedation and analgesia are covered, to include general principles and specific agents for its use in the emergency department. PMID- 17400073 TI - Closure techniques. AB - There are many techniques available for the closure of traumatic wounds in emergency departments. Each method has its own unique benefits and limitations that must be considered in each case individually. The best technique provides durability, simplicity, excellent cosmetic outcome, and overall patient satisfaction. Unfortunately, at this time no single method is superior for all situations. Emergency physicians must be familiar with multiple techniques to provide the best outcomes for their patients. PMID- 17400074 TI - Advanced laceration management. AB - Many lacerations seen in the emergency department setting require specific management based on anatomic location. Lacerations of the fingertip, ear, nose, lip, tongue, and eyelid can be complex and require advanced management techniques. Many can be primarily treated by emergency clinicians; however, it is important for the clinician to know when consultation is appropriate for treatment by a specialist. Current literature recommendations are presented for initial management, methods of repair, technical tips to facilitate repair, appropriate consultation, and postoperative care for these complex lacerations. PMID- 17400075 TI - Emergency management of difficult wounds: part I. AB - Emergency physicians treat millions of wounds annually in the United States. In 2004 approximately 6.4 million open wounds were seen in United States emergency departments, representing approximately 5.8% of all visits. This article discusses difficult traumatic wounds that emergency physicians encounter on a daily basis, including mammalian bites, puncture and high-pressure wounds, and crush injuries, with special emphasis given to individuals at high risk for infectious complications. Information is provided on epidemiology, pathophysiology, management, and treatment of various complicated wounds. The authors highlight available guidelines, provide the best evidence available, and provide recommendations when data are limited. PMID- 17400076 TI - Emergency management of difficult wounds: part II. AB - Difficult wounds constitute a significant amount of the morbidity and mortality emergency physicians face on a daily basis. There are specific traumatic and atraumatic wounds that are difficult to manage and have a high risk of complications. Emergency physicians must be able to identify these high-risk wounds and patients and take steps to mitigate further morbidity and mortality. PMID- 17400077 TI - Management of burn wounds in the emergency department. AB - This article makes some introductory comments on the histology of the skin and the pathophysiology of burn injury as these topics pertain to the estimation of the depth of the burn injury. The definition of a major burn and the salient points of its treatment are covered. In addition, some general comments are made about several special injuries for which burn center referral usually is sought. Finally, guidance is given in the selection and treatment of patients who have burns that may be treated on an outpatient basis. PMID- 17400078 TI - Postcare recommendations for emergency department wounds. AB - Diligent posttreatment wound care management undoubtedly will improve wound outcome and patient satisfaction. There are limited recommendations in the literature to guide management plans. Nevertheless patients must receive specific instructions to complete wound care. These instructions should include whether a dressing is indicated, which dressing should be used, the duration of use, and the method of application. The plan must explain clearly the reasons for returning for further medical attention, for follow-up, for routine removal of sutures/staples, and an earlier return for possible concerns of infection or dehiscence. Preprinted discharge instruction sheets are useful, and illustrations can be helpful. PMID- 17400079 TI - Use of appropriate antimicrobials in wound management. AB - The primary goal of wound management is to achieve a functional closure with minimal scarring. Preventing infection is important to facilitate the healing process. Most simple, uncomplicated wounds do not need systemic antibiotics but benefit from the use of topical antibiotics. Judicious use of antibiotics reduces unnecessary adverse events and helps reduce the development of resistance. Although antibiotics can help reduce infection risk and promote healing, they are not a substitute for good local wound care, in particular irrigation and surgical debridement. This article reviews the role of antibiotics in emergency department wound management. PMID- 17400080 TI - Hyperbaric oxygen therapy in the treatment of open fractures and crush injuries. AB - This article focuses on the use of hyperbaric oxygen therapy (HBOt) in the treatment of open fractures and crush injuries. Based on the clinical evidence and cost analysis, medical institutions that treat open fracture and crush injuries are justified in incorporating HBOt as a standard of care. Both Medicare and Undersea and Hyperbaric Medical Society guidelines list crush injuries as an approved indication for HBOt. Emergency physicians should familiarize themselves with this emerging treatment modality because of their role in the early management of these injuries. PMID- 17400081 TI - Reducing risk in emergency department wound management. AB - Although substantial dollar amounts are not involved, wound-care litigation constitutes a significant number of lawsuits to emergency medicine physicians, resulting in an increased drain on the physician's time and exposing the physician to all the psychosocial effects involved in the medicolegal process. The procedures outlined in this article-paying attention to wound-care principles, involving patients in the medical decision-making process, and ensuring appropriate medical follow-up-can, it is hoped, reduce the incidence of medical claims. PMID- 17400082 TI - Emergency management of chronic wounds. AB - As America's emergency departments witness an increase in care provided to an aging population, the emergency physician increasingly evaluates and treats manifestations of chronic disease. Nonhealing wounds are often a presenting manifestation of chronic disease. They are a source of pain and disability for this population. Emergency physicians should possess a fundamental knowledge in the management of chronic wounds. This article familiarizes the emergency physician with the epidemiology of chronic wounds, the physiology of tissue repair, the pathophysiology involved in wound healing failure, the common types of chronic wounds, and specific management strategies. PMID- 17400083 TI - Imaging modalities in wounds and superficial skin infections. AB - Open wounds and lacerations are the most commonly encountered problems in emergency medicine. Detection and removal of a foreign body is essential to avoid the many complications of a retained foreign body, which may include infection, inflammation, allergic reaction, and disability. Currently, there are several imaging modalities that the emergency medicine provider may use to aid in foreign body detection and wound management. Techniques and appropriate use of these imaging modalities for foreign bodies and soft tissue infections are discussed in this article. PMID- 17400084 TI - Occlusive wound dressings in emergency medicine and acute care. AB - One of the great misconceptions in wound care is that a wound heals best when permitted to form a dry scab. By contrast, moisture has repeatedly been shown to significantly accelerate wound healing. Emergency physicians and other acute care providers are encouraged to incorporate occlusive moisture-retentive dressings into their regular practice to expedite healing, reduce pain and scarring, improve wound care convenience and patient compliance, and minimize wound contamination and infection. PMID- 17400085 TI - Failure of the American College of Chest Physicians-1A protocol for lovenox in clinical outcomes for thromboembolic prophylaxis. AB - A total of 290 consecutive patients who underwent total hip and total knee arthroplasty were prospectively entered into a clinical anticoagulation trial using a 10-day course of Lovenox with the American College of Chest Physicians-1A guidelines. Major complications occurred in 9% of patients; symptomatic deep vein thrombosis occurred in 9 (3.8%) patients, and nonfatal pulmonary embolism in 3 (1.3%) patients. Complications included 4.7% readmissions, 3.4% return to the operating room for wound incision and drainage, 5.1% prolonged hospitalization (wound drainage), and 3.4% injection site complications. Wound drainage of more than 7 days was predictive of readmission and wound reoperation. A body mass index of more than 35 was predictive of prolonged wound drainage. Return to the operating room for wound complications occurred 3x more frequently with the use of Lovenox than in our previous study using warfarin. Surgical site complications requiring readmission or reoperation should be considered "major" complications. PMID- 17400086 TI - A randomized, parallel-group, open-label trial of recombinant human erythropoietin vs preoperative autologous donation in primary total joint arthroplasty: effect on postoperative vigor and handgrip strength. AB - This randomized trial assessed the effect of recombinant human erythropoietin (EPO) vs preoperative autologous donation (PAD) on postoperative vigor and handgrip strength in patients undergoing primary total joint arthroplasty. Adults with baseline hemoglobin level of 11 to 14 g/dL received EPO (600 IU/kg once weekly for 4 doses, n = 130) or PAD (n = 121) before primary, unilateral hip or knee arthroplasty. Mean changes in vigor score and handgrip strength from baseline were not significantly different between treatment groups. Multivariate analyses found a significant treatment effect favoring EPO over PAD for vigor, but not for handgrip strength. Patients in the EPO group had higher hemoglobin levels and required fewer transfusions. Both treatments were well tolerated. Additional study is needed to elucidate the influence of blood management strategies on postoperative vigor. PMID- 17400087 TI - Computer-assisted gap equalization in total knee arthroplasty. AB - This in vitro biomechanical study compared a conventional balancing technique in knee arthroplasty to a technique using computer assistance. The experimental technique used a soft tissue tensioner instrumented with computer-monitored load cells to quantify soft tissue tension. To assess outcome, a tibial load transducer measured medial and lateral compartment forces and tibial rotation. An electromagnetic tracking system quantified knee position under simulated muscle loading. The computer-assisted technique improved knee balance before insertion of components. However, once components were implanted, there was no difference in knee load balance between the 2 techniques. No correlation was shown between compartmental load balance and tibial rotation or mechanical axis misalignment. Although computer-assisted surgery may improve technical accuracy, further work is necessary to achieve an optimal final knee load balance. PMID- 17400088 TI - The inferior vena cava filter is effective in preventing fatal pulmonary embolus after hip and knee arthroplasties. AB - Thromboembolic disease is a relatively common and potentially devastating complication of joint arthroplasty. Mechanical and chemical prophylaxes are effective in reducing the incidence of this complication. Inferior vena cava (IVC) filters have been used to prevent the propagation and/or migration of venous emboli into the pulmonary circulation. This article reports on a cohort of joint arthroplasty patients either with confirmed pulmonary embolism or at high risk of developing pulmonary emboli in whom an IVC filter was implanted. Using the institutional database, 95 joint arthroplasty patients who received an IVC filter were identified. The IVC filter was effective in preventing fatal pulmonary emboli in all patients. The IVC filter is a valuable and effective modality in preventing a fatal outcome of thromboembolism in patients with established thromboembolism. PMID- 17400089 TI - A randomized, prospective study of primary total knee components designed for increased flexion. AB - This prospective, randomized study reports early results for range of motion (ROM) after primary total knee arthroplasty. Group A received standard posterior stabilized (PS) implants. Group B had PS implants designed for increased flexion. Average preoperative ROM was 2 degrees to 121 degrees for group A and 1.8 degrees to 122 degrees for group B. At 12-month follow-up, average ROM for group A was 0.6 degrees to 120 degrees as compared with 0.6 degrees to 133 degrees for group B (P < .05). Significantly more patients in group B flexed more than 135 degrees (P < .05). Furthermore, 12 patients (48%) in group A failed to return to preoperative ROM versus 2 patients (8%) in group B (P < .05). Although long-term follow-up is desirable, these early results support the use of PS implants designed for increased flexion. PMID- 17400090 TI - Skin incision translation in total knee arthroplasty: the difference between flexion and extension. AB - Scar location and point loading of the scar after total knee arthroplasty are common postoperative concerns for patients and physicians. A frequent problem is discomfort associated with the incision. We evaluated 40 knees to better understand the association of the tibial tubercle with the overlying skin in both flexion and extension. Our results revealed lateral translation of the skin overlying the tibial tubercle in 100% of our volunteers as the knee is flexed and the tibia internally rotates. The average amount of lateral translation measured 9.7 mm. This could have a significant impact on point loading of the scar if an anterior midline incision for total knee arthroplasty is planned in extension. PMID- 17400091 TI - Clinical outcome of total hip arthroplasty using the normalized and proportionalized femoral stem with a minimum 20-year follow-up. AB - Currently, there are several femoral stem designs available for use, but few have an extended track record. We have previously reported on 10- and 15-year outcome studies of total hip arthroplasty (THA) using a cemented normalized and proportionalized femoral stem from a single surgeon series. This is a follow-up study reporting the minimum 20-year outcome of this femoral stem design. The study began with THA performed in a consecutive series of 184 patients; stem fixation was achieved using first-generation cementing techniques. The overall early complication rate was 10%. There were 23 patients (31 hips) who had been followed-up for a minimum 20-year period (average 21.3 years). Mean d'Aubigne and Postel scores improved from 5.9 to 11.3; mean Harris hip scores improved from 43.8 to 92.8. Kaplan-Meier survivorship was 93% at 20 years (95% confidence interval); there were no stem failures. The use of a cemented normalized and proportionalized femoral stem in primary THA provides satisfactory long-term clinical and radiological outcomes in patients. PMID- 17400092 TI - Ten-year survivorship of the Anatomique Benoist Girard I total hip arthroplasty. AB - The Anatomique Benoist Girard (ABG) I total hip arthroplasty has been widely used in Europe since 1989. Two recent midterm reports have shown high rates of component failure secondary to polyethylene wear. To further investigate this issue, our study prospectively follows 157 consecutive primary total hip arthroplasties using the ABG I prosthesis at an average follow-up of 10 years. Clinical outcomes have been excellent, although the incidence of thigh pain was 7.2%. Radiographic evaluation of the cup showed eccentric polyethylene wear in 93.6% of the patients. The average polyethylene linear wear was 2.40 mm, whereas the average annual wear rate was 0.25 mm/y. There were 7 hips (5.6%) with severe periacetabular osteolysis. Nine revisions were performed for loosening or wear. The overall component survival rate was 92.6%. The overall survival rate was 92.6% (confidence interval, 78.55-96.47) for the acetabular component and 96.3% (confidence interval, 85.67-100) for the femoral component. Although the clinical and radiographic results of the ABG I total hip arthroplasty in this series have been acceptable, high rates of polyethylene wear, most likely related to sterilization by gamma-irradiation in air, have been the limiting factor in the long-term success of this design. PMID- 17400093 TI - Cementless hemispheric hydroxyapatite-coated sockets for acetabular revision. AB - This study describes our experience with a hydroxyapatite-coated uncemented hemispherical component used for revision of 72 patients with aseptic loosening of the acetabular component. Preoperative Paprosky classification of the acetabular defects was 1 type I, 35 type II, and 36 type III; according to American Academy of Orthopaedic Surgeons, 14 segmental, 10 cavitary, and 48 combined. The mean follow-up was 7.6 years (range, 5.0-13.0). Complications were seen in 7 cases (9.6%). Seventy acetabular components (97.2%) showed bone ongrowth on the radiographs. The survival rate was 90.8% after a mean follow-up of 7.6 years (range, 5.0-13.0) when revision for any reason is the end point (confidence interval, 80.5%-100%) and 98.1% (confidence interval, 94.5%-100%) when aseptic loosening of the cup is the end point. We conclude that hydroxyapatite-coated cups for acetabular revision show promising results. PMID- 17400094 TI - The constrained acetabular component for hip instability. AB - One hundred four hips in 107 patients undergoing revision arthroplasty of the hip were identified at risk of dislocation and treated with the constrained cup. Radiostereometric analysis was performed to assess prosthesis migration. Mean follow-up was 3.0 years (range, 2.0-4.8). At last review, 19 patients had died and 6 were lost to follow-up. There were 5 revisions for cup loosening and a further 4 with radiographic evidence of loosening. There were 3 dislocations and 3 dissociations in 5 patients. Radiostereometric analysis demonstrated that cup migration at 24 months was up to 0.82 mm of translation and 1.58 degrees of rotation. Our results confirm that the constrained acetabular component is a highly effective option for the treatment for patients with instability of the hip. The aseptic loosening rate was higher than previously reported. PMID- 17400095 TI - Magnetic resonance imaging analysis of kinematics in osteoarthritic knees. AB - Kinematics in osteoarthritic knees may be impeding efforts to reproduce "normal" knee kinematics in the prosthetic knee. Fourteen subjects with unilateral symptomatic knee osteoarthritis performed a supine leg press from 0 degrees to 90 degrees flexion against a 150-N load. The tibiofemoral contact pattern was recorded for both knees using sagittal T1-weighted magnetic resonance imaging. Severity of osteoarthritis ranged from Kellgren Lawrence grade 2 to 4 in the symptomatic knees and from 0 to 3 in the contralateral knees. Contact in the lateral and medial compartments of osteoarthritic knees was more anterior on the tibial plateau than healthy knees, both in knee extension and to 90 degrees flexion (P < .01). This anterior contact pattern was associated with severity of osteoarthritis (P < .01). PMID- 17400096 TI - Shear properties of bilaminar polymethylmethacrylate cement mantles in revision hip joint arthroplasty. AB - Although cement-within-cement revision arthroplasty minimizes the complications associated with removal of secure PMMA, failure at the interfacial region between new and old cement mantles remains a theoretical concern. This article assesses the variability in shear properties of bilaminar cement mantles related to duration of postcure and the use of antibiotic cements. Bilaminar cement mantles were 15% to 20% weaker than uniform mantles (P < .001) and demonstrated variability in shear strength related to duration of postcure of the freshly applied cement (P < .001). The use of Antibiotic Simplex did not significantly influence interfacial cement adhesion (P = .52). Interfacial adhesion by mechanisms other than mechanical interlock plays a significant role in the bond formed between new and old PMMA cements, with an important contribution by diffusion-based molecular interdigitation. In the presence of a secure cement bone interface, we recommend cement-within-cement revision techniques in suitable patients. PMID- 17400097 TI - In vivo contact pressures in total knee arthroplasty. AB - This study compared the in vivo femoropolyethylene contact pressures generated in fixed-bearing total knee arthroplasty (TKAs) with those in mobile-bearing TKAs. In vivo kinematics obtained from a 2-dimensional to 3-dimensional registration technique and soft tissue locations derived from computed tomographic scans were entered into a 3-dimensional inverse dynamics mathematical model to determine the in vivo bearing contact forces. The contact areas were obtained from the assembly of computer-aided design models of the components. The contact pressure was defined as the ratio of the contact forces to the contact areas. The results indicate that the in vivo contact pressures in each TKA are greater for the medial condyle than for the lateral condyle. The ability of the mobile-bearing TKA to rotate maintains higher femoropolyethylene contact, resulting in lesser contact pressures, as compared with the fixed-bearing TKA. PMID- 17400098 TI - Intramedullary canal pressure distribution: an experimental parametric study. AB - Intramedullary instrument/device insertion during total joint replacement can generate high intramedullary pressures, with potential for clinical fat embolism syndrome. We evaluated effects of process parameters on intramedullary pressure generated when hammering an implant into a simulated femur analogue. Lower implant insertion speed, lower hammering force, a rubber- compared with steel tipped hammer and a larger synthetic bone-to-implant radial gap reduced intramedullary pressure generated. Number of hammer strikes required to insert an implant increased with increasing marrow viscosity, whereas intramedullary pressure decreased with increasing viscosity. Significant intramedullary pressure reduction is obtained by applying more strikes of lower peak force using a rubber hammer tip, as opposed to fewer larger strikes. Caution should be exercised when instrumenting the distal femur, where viscosity is low and very high intramedullary pressures can be generated. PMID- 17400099 TI - Preheating acrylic bone cement powder is not recommended for all brands. AB - One strategy that has been suggested for reducing the operating room time for cemented total joint arthroplasties-and, hence, for reducing the total cost of these procedures-is to accelerate the polymerization of the acrylic bone cement by preheating the powder to 65 degrees C. We quantified the effect of preheating the cement powders on the fracture toughness and fatigue life of 3 cement brands that are widely used in clinical practice. The results suggest judicious selection of cements whose powders are to be preheated for use in cemented arthroplasties. PMID- 17400100 TI - Simulated normal gait wear testing of a highly cross-linked polyethylene tibial insert. AB - Polyethylene wear can compromise long-term performance of total knees. We investigated articular and backside wear rate of tibial inserts made from conventional vs highly cross-linked polyethylenes. The inserts were tested on a knee simulator at a frequency of 1.1 Hz using normal gait kinematics with a 7 degrees posterior slope for 7 million cycles. Gravimetrically, the combined articular and backside wear rate was 23 +/- 6 and 4.6 +/- 2 mm(3) per million cycles for the conventional and cross-linked inserts, respectively. We independently measured articular surface wear rates, which were 15 +/- 4 and 5 +/ 2 mm(3) per million cycles for conventional and cross-linked inserts, respectively. Electron beam cross-linked and subsequently melted polyethylene reduced the wear rate of polyethylene on articular and backside surfaces in a cruciate-retaining total knee design when tested on a knee simulator under simulated normal gait. PMID- 17400101 TI - Computer-assisted fluoroscopic navigation system for removal of distal femoral bone cement in revision total hip arthroplasty. AB - Removal of the femoral bone cement in revision total hip arthroplasty with a high powered drill or burr potentially has a risk of damage to the bone, resulting in perforation and fracture of the femur. Recently, we have used a computer-assisted fluoroscopic navigation system for the revision of cemented total hip arthroplasty with a high-powered burr and completely removed the distal femoral bone cement with no complications in 6 cases. Thus, a computer-assisted fluoroscopic navigation system is a useful tool for the improvement of the surgical technique in revision total hip arthroplasty. PMID- 17400102 TI - Periprosthetic fracture between a constrained total knee arthroplasty and a long stem total hip arthroplasty: treatment with a novel device. AB - Supracondylar fracture of the femur occurred in a patient with a well-fixed constrained condylar total knee arthroplasty and an 8-in total hip femoral revision component. This unusual fracture was treated with a novel interpositional femoral intramedullary device that connected the total knee to total hip arthroplasty. The result was good at 5 years, with healed fracture and no loosening or osteolysis of either component. PMID- 17400103 TI - Extravascular compression of the femoral vein due to wear debris-induced iliopsoas bursitis: a rare cause of leg swelling after total hip arthroplasty. AB - We present a patient with unilateral, spontaneous, late leg swelling that developed 4 years after total hip arthroplasty. The etiology was the compression of the internal iliac vein by a voluminous iliopsoas bursitis caused by polyethylene debris. The expansive lesion was detected by ultrasound, arthrography, and magnetic resonance imaging. An ultrasound-guided aspiration provided transient relief of the patient's symptoms. The patient later required surgical excision through an abdominal approach. A second recurrence was detected and treated with revision surgery. We present the diagnosis and the treatment of this rare cause of late, unilateral leg swelling after total hip arthroplasty together with a review of the literature. PMID- 17400104 TI - Management of a multidrug-resistant Pseudomonas aeruginosa infected total knee arthroplasty using colistin. A case report and review of the literature. AB - Multidrug-resistant infections present a serious clinical and therapeutical problem. Colistin is an old-used polymyxin with rather poor pharmacokinetic profile and a remarkable nephrotoxicity. However, the emergence of multidrug resistant bacteria has recently led to the increased use of colistin as a potentially available therapy. This article presents a 75-year-old diabetic woman with an early onset total knee arthroplasty infection by a multidrug-resistant Pseudomonas aeruginosa bacterial isolate that was managed successfully with surgical removal of the knee prosthesis, antibiotic impregnated cement and intravenous administration of colistin for 6 weeks, and second stage revision knee surgery. Two years later, laboratory and imaging studies showed no evidence of recurrence of infection. PMID- 17400105 TI - Failure of a metal-reinforced tibial post in total knee arthroplasty. AB - Failures of the polyethylene tibial post in posterior stabilized total knee arthroplasty are usually associated with pain and knee instability. We report an unusual presentation of a tibial post failure that occurred after the polyethylene insert was revised in a posterior-stabilized total knee. The tibial post on the revised insert broke off, exposing a metal reinforcing pin. During the resulting posterior subluxations of the tibia, the metal pin articulated against the cam on the femoral component, resulting in the generation of metallic debris in the knee joint. PMID- 17400106 TI - Alumina femoral head fracture in uncemented total hip arthroplasty with a ceramic sandwich cup. AB - In total hip arthroplasty, insertion of a UHMWPE-coated ceramic sandwich liner dramatically reduces any risk related to the stiffness of the ceramic-ceramic coupling. We present a case of an alumina ceramic head fracture with a ceramic sandwich cup. The fracture occurred 16 months after the initial operation, without trauma. Impingement between the neck and posteroinferior portion of the liner rim had occurred. This was related to the traditional cross-legged sitting position of Koreans, which caused impingement between the neck and liner rim. Consequently, the taper became loose, and the resulting force bent the taper and fractured the head. We examined a peculiar case of ceramic femoral head fracture after implantation of an uncemented total hip arthroplasty with a ceramic sandwich cup and investigated the underlying cause. PMID- 17400107 TI - Compartment syndrome and popliteal vascular injury complicating unicompartmental knee arthroplasty. AB - Popliteal vascular injury and the compartment syndrome of the leg are rare but important complications of knee arthroplasties. Early diagnosis and treatment are of paramount importance in preventing the devastating complications of these conditions. To our knowledge, these complications have not been reported previously after unicompartmental knee arthroplasty in the literature. Low level of suspicion may delay the diagnosis, as popliteal vascular injury and compartment syndrome are not well recognized as possible complications of unicompartmental knee arthroplasty. PMID- 17400108 TI - Fatal cerebral emboli in the absence of a cardiac arterial-venous shunt: case report. AB - Cerebral embolism in patients undergoing total joint arthroplasty has an incidence rate of 0.5%. In those cases where cerebral emboli were confirmed, a venous-to-arterial circulation shunt, most commonly a patent foramen ovale, was identified. We report a case of severe cerebral fat emboli during cementless total hip arthroplasty in the absence of an intracardiac defect. PMID- 17400110 TI - Sonographers' communication pressure to meet standards? No problem. PMID- 17400111 TI - Evaluation of longitudinal and radial two-dimensional strain imaging versus Doppler tissue echocardiography in predicting long-term response to cardiac resynchronization therapy. AB - INTRODUCTION: Cardiac resynchronization therapy (CRT) leads to hemodynamic and clinical improvement in patients with heart failure. This study compares the new technique of 2-dimensional (2D) strain imaging with Doppler tissue imaging (DTI) in the prediction of the long-term response to CRT. METHODS: In all, 38 patients with heart failure (New York Heart Association II-III, QRS > 120 milliseconds, left ventricular ejection fraction < 0.35) received CRT and echocardiographic evaluation with a mean follow-up of 9.4 months. RESULTS: Of the patients, 47.4% were hemodynamic responders to long-term CRT. In the responder group, the maximum delay in the longitudinal and radial 2D strain in the basal segments and the maximum delay in the DTI peak systolic myocardial velocities but not DTI strain decreased. In receiver operating characteristic analysis, the baseline delay of DTI peak velocities predicts improvement after CRT, whereas baseline 2D- and DTI strain measurements fail to predict a long-term response to CRT. CONCLUSION: Although there is a significant decrease in longitudinal and radial 2D strain derived delays after CRT, the resulting predictive value is not superior to DTI. PMID- 17400112 TI - Assessment of segmental myocardial viability using regional 2-dimensional strain echocardiography. AB - We determined whether 2-dimensional strain echocardiography can identify viable from infarcted myocardium in a rat ischemia-reperfusion model. A total of 16 male Sprague-Dawley rats underwent left anterior descending coronary artery occlusion for 12 or 30 minutes followed by 60-minute reperfusion. Short-axis 2-dimensional strain echocardiography was performed at the mid-ventricle 60 minutes post reperfusion. Post-sacrifice, triphenyl tetrazolium chloride was infused to the coronary circulation. Regional end-systolic radial and circumferential strain, and time to peak strain, were measured using software in all 96 segments and correlated with areas of infarct in corresponding histologic slices. Segments with greater than 50% area of infarct had lower end-systolic radial and circumferential strain and longer time to peak strain versus areas with 50% or less strain or no infarct. Extent of infarct correlates with radial and circumferential strain. End-systolic radial strain less than 2% has 88% sensitivity and 95% specificity for detecting infarcted area greater than 50%. Two-dimensional strain echocardiography-derived strain is useful in distinguishing infarcted from viable myocardium. PMID- 17400113 TI - Transient ischemic left ventricular cavity dilation is a significant predictor of severe and extensive coronary artery disease and adverse outcome in patients undergoing stress echocardiography. AB - BACKGROUND: Transient ischemic dilation (TID) of the left ventricle on stress redistribution thallium-201 scintigraphy is a marker of severe and extensive coronary artery disease (CAD), and associated with an adverse outcome. The significance of transient ischemic dilation during stress echocardiography is not well defined. METHODS: We assessed 155 patients undergoing stress echocardiography (61% treadmill exercise, 39% dobutamine) with confirmed follow up (mean 2.8 +/- 1.1 years) for hard events (myocardial infarction, n = 14, and cardiac death, n = 9). RESULTS: Normal limits for TID ratio were developed using data from 39 patients with a low likelihood (<5%) of CAD and normal stress echocardiography study findings. The criteria for abnormality was developed based on data from 116 patients who underwent coronary angiography after and within 3 months of an abnormal ischemic stress echocardiography study result. For normal limits, receiver operating characteristic curve analysis showed that abnormal TID ratio values corresponded to left ventricular volume ratios greater than 1.17 (mean +/- 2SD). TID assessment using these criteria for abnormality showed high sensitivity (100%) and moderate specificity (54%) for detection of severe and extensive angiographic CAD. Patients with TID had a greater extent and severity of stress induced wall-motion abnormalities, higher peak wall-motion score index, and worse prognosis than patients without TID. CONCLUSIONS: TID during stress echocardiography is a sensitive marker of severe and extensive angiographic CAD and is associated with a high risk of cardiac events (19.7%/y event rate). PMID- 17400114 TI - Usefulness of left atrial volume for the differentiation of normal from pseudonormal diastolic function pattern in patients on hemodialysis. AB - BACKGROUND: Discrimination of normal from pseudonormal mitral flow is challenging in clinical practice, especially in a milieu of acute plasma volume changes, such as in patients undergoing hemodialysis (HD). Left atrium (LA) enlargement has been recognized as a marker of the duration of left ventricular diastolic dysfunction. We hypothesize that LA volume index (LAVi) may be useful to differentiate normal from pseudonormal left ventricular filling pattern in this scenario. METHODS: Fifty clinically stable patients undergoing HD (of 138 evaluated in a routine examination) in sinus rhythm who had an early to atrial transmitral velocity ratio between 1 and 2 on Doppler echocardiography were selected and divided into normal (n = 28) and pseudonormal (n = 22) groups based on mitral annulus tissue Doppler and pulmonary venous flow data. Receiver operating characteristic curves for LAVi and other echocardiographic parameters were generated to compare the discriminating power between normal and pseudonormal mitral flow. RESULTS: The mean LAVi for patients with normal mitral flow was 24 +/- 6 versus 48 +/- 16 mL/m(2) in those with pseudonormalization (P < .001). LAVi had the best overall performance in comparison with several echocardiographic parameters (area under the curve 0.94, 95% confidence interval 0.84-1.0, P < .001). LAVi greater than 35 mL/m(2) was 91% sensitive and 89% specific for the detection of pseudonormalization in patients undergoing HD. CONCLUSION: Enlargement of LA reflects the chronicity and burden of left ventricular diastolic dysfunction in stable patients undergoing HD with early to atrial transmitral velocity ratio apparently normal on conventional Doppler echocardiography. LAVi is a practical and useful parameter to discriminate normal from pseudonormal mitral flow. PMID- 17400115 TI - The relationship between aortic stiffness and cardiac function in patients with obstructive sleep apnea, independently from systemic hypertension. AB - OBJECTIVE: We sought to evaluate aortic function and its relationship with left ventricular (LV) function in patients with obstructive sleep apnea syndrome (OSAS), with or without systemic hypertension. METHODS: A total of 29 patients with OSAS, 28 patients with systemic hypertension without OSAS, and 19 patients having both disorders were chosen as a study group, and 29 participants without these two disorders were taken as the control group; none of the patients had a history of cardiac disease. All of them underwent echocardiography and polysomnography. Aortic strain and distensibility were calculated from the aortic diameters measured by echocardiography and blood pressure obtained by cuff sphygmomanometer. Cardiac functions were determined using echocardiography comprising standard 2-dimensional and conventional Doppler and Doppler tissue imaging. RESULTS: Aortic strain (mean +/- SD; 6.1 +/- 2.7% vs 6.4 +/- 2.4%, 4.7 +/- 1.8%, and 13.7 +/- 4.5%, P < .001, respectively) and distensibility (mean +/- SD 2.8 +/- 1.6 vs 2.5 +/- 0.9, 1.7 +/- 0.7, and 6.2 +/- 3.2 10(-6) cm(2) dyn(-1)P < .001, respectively) were significantly decreased in the patient groups (OSAS, hypertensive, and OSAS and hypertensive) compared with the control group. LV long axis and diastolic functions were also impaired in the study group. There were good correlations between the aortic stiffness and LV function parameters. In a multivariate analysis, aortic strain was the parameter found to be most strongly associated both with the Doppler tissue imaging mean peak early/late diastolic velocity ratio and the LV mitral lateral annular plane systolic excursion. CONCLUSION: Aortic stiffness parameters (aortic strain and distensibility), LV systolic long-axis function indicators (mitral lateral annular plane systolic excursion, peak systolic myocardial velocity at lateral annulus, peak systolic myocardial velocity at septal annulus), and Doppler tissue imaging-derived LV diastolic indices were found abnormal in patients with OSAS, hypertension, or both. Thus, subclinical cardiac dysfunction has been found in patients with OSAS independent from the systemic hypertension. PMID- 17400116 TI - Efficient quantification of the left ventricular volume using 3-dimensional echocardiography: the minimal number of equiangular long-axis images for accurate quantification of the left ventricular volume. AB - For quantification of the left ventricular volume from 3-dimensional echocardiograms a number of cross-sectional images are used. The goal of this study was to determine the minimum number of long-axis images necessary for accurate quantification of the left ventricular volume. A strong correlation was observed between volumes obtained from magnetic resonance imaging and 3 dimensional echocardiography using 16 equiangular images (r = 0.99; y = 0.95x + 3.3 mL; standard error of the estimate = 7.0 mL; N = 30). Comparison of these results with random subsets showed a significant difference for volumes obtained with 4 and 2 equiangular images (P < .005). However, when the subsets were selected to target the eccentric region of the endocardial border this was only the case for subsets of two images (P < .001). This study demonstrates that accurate left ventricular volume quantification can be performed with as little as 8 equiangular long-axis images. By selecting the correctly oriented image set, this number can even be brought down to 4, which will further reduce the analysis time. PMID- 17400117 TI - Dynamic change in mitral annular area and motion during percutaneous mitral annuloplasty for ischemic mitral regurgitation: preliminary animal study with real-time 3-dimensional echocardiography. AB - We used a novel 3-dimensional (3D) echocardiographic technique to evaluate the impact of a coronary sinus-based percutaneous transvenous mitral annuloplasty (PTMA) on dynamic changes in mitral annular geometry and motion during the cardiac cycle in 8 sheep with ischemic mitral regurgitation. Using real-time 3D echocardiographic data before and after PTMA, 10 points along the saddle-shaped annulus were identified. For every 3D volume/frame during a cardiac cycle, we assessed mitral annular area and excursion defined as the traveling distance of the annular center. The PTMA device reduced both minimum and maximal mitral annular area (9.5 +/- 0.9-7.0 +/- 0.6 and 12.8 +/- 1.3-9.8 +/- 1.5 cm(2), P < .001 for both, respectively) with reduction of mitral regurgitation jet area (5.1 +/- 2.3-1.2 +/- 0.8 cm(2), P < .001), whereas it did not significantly impair mitral annular excursion amplitude (8.3 +/- 1.1-7.0 +/- 1.9 mm, P = .13). This 3D echocardiographic method noninvasively enabled dynamic study of mitral annular geometry and motion with quantitative analysis of the impact of PTMA. PMID- 17400118 TI - Proximal flow convergence region as assessed by real-time 3-dimensional echocardiography: challenging the hemispheric assumption. AB - OBJECTIVE: Traditionally, a hemispheric assumption for the proximal flow convergence region (PFCR) is used when calculating mitral regurgitant (MR) effective orifice area (EROA). However, 2-dimensional (2D) echocardiography limits evaluation of the complete PFCR contour. Real-time 3-dimensional (3D) echocardiography (RT3D) allows direct assessment of the true PFCR contour. We hypothesized that the PFCR contour is not necessarily hemispheric, but rather hemielliptic, and aimed to apply a hemielliptic calculation, based on the 3D contour of the PFCR for more accurate MR quantification. METHODS: In all, 50 patients with MR underwent RT3D to characterize PFCR contour as hemispheric or hemielliptic. MR EROA by RT3D-derived PFCR was calculated using a hemielliptic formula using 3D data. The 2D EROA was computed using standard hemispheric assumption. EROAs calculated from 2D and RT3D data were compared with quantitative Doppler EROA (mitral inflow--aortic outflow/MR time-velocity integral), used as an independent comparison. RESULTS: Only 1 of 50 patients (2%) had a hemispheric PFCR contour by RT3D. The remaining had hemielliptic PFCR contours. Compared with Doppler method, 2D echocardiography significantly underestimated EROA (0.34 +/- 0.14 vs 0.48 +/- 0.25 cm(2), P < .001). RT3D EROA was not significantly different from Doppler EROA (0.52 +/- 0.17 vs 0.48 +/- 0.25, P = not significant). Of 33 patients with Doppler EROA greater than 0.3 cm(2) (> or =moderate-severe MR), 45% (15 of 33) were underestimated as having mild to moderate MR by 2D EROA. CONCLUSIONS: The true PFCR contour as shown by RT3D is generally not hemispheric but hemielliptic, tracking the orifice contour. Based on this 3D shape, a hemielliptic approach can be used for practical clinical application with improved MR quantification. PMID- 17400119 TI - Mitral annular remodeling with varying degrees and mechanisms of chronic mitral regurgitation. AB - HYPOTHESES: Degree of mitral annular remodeling is directly associated with severity of chronic mitral regurgitation (MR). Mitral annular remodeling occurs in a symmetric fashion, regardless of MR severity. In addition to MR severity, MR mechanism plays a significant role in mitral annular remodeling. BACKGROUND: Limited data exists on mitral annular remodeling in patients with MR. Identification of annular changes may be important in aiding surgical repair. METHODS: Mitral annular dimensions (anteroposterior, intercommissural, surface area, and circumference) were measured in end systole and diastole using 3 dimensional reconstructive software in 83 patients: trace to no MR (23), mild MR (15), moderate MR (26), and severe MR (19). Annular sphericity indices were determined by dividing intercommissural by anteroposterior dimensions. Patients were further subgrouped by mechanism of MR. RESULTS: With increasing MR severity, there was a corresponding increase in all annular measurements, most pronounced in the anteroposterior dimension, circumference, and area. Larger increases were seen in patients with prolapse/flail and dilated mechanisms. Furthermore, the mitral annulus became more circular (sphericity index approached 1.0) with increasing MR severity. Patients with prolapse/flail mechanisms exhibited normal left ventricular volumes despite significant annular enlargement. CONCLUSIONS: Mitral annular remodeling is directly associated with MR severity and occurs in an asymmetric fashion, yet is not limited to one region of the annulus. Mechanism of MR plays a significant role in annular remodeling. Annular remodeling can occur independently of left ventricular remodeling. PMID- 17400120 TI - Echocardiography-based spectrum of severe tricuspid regurgitation: the frequency of apparently idiopathic tricuspid regurgitation. AB - BACKGROUND: The cause of tricuspid valve (TV) regurgitation (TR) occasionally remains unclear. The objectives of our study were to define the causal spectrum of severe TR diagnosed by echocardiography at a tertiary medical center and to assess the relative frequency and determine the clinical and echocardiographic characteristics of TR without an apparent cause (idiopathic TR). METHODS: Consecutive patients with severe TR were identified by the echocardiography laboratory computerized database. The echocardiographic reports of all patients were reviewed and the causes of TR were determined. The echocardiographic studies and medical charts were reviewed in patients without an obvious cause of TR. RESULTS: Of 242 consecutive patients diagnosed with severe TR, organic TV disease was evident in 23 patients (9.5%) and significant pulmonary hypertension (estimated pulmonary artery systolic pressure > 50 mm Hg) in an additional 157 patients (64.9%). After further excluding patients with various confounding factors, possibly associated with occult organic TV disease or significant pulmonary hypertension, 23 patients (9.5%) had severe TR without an apparent cause. Of these, TV coaptation appeared relatively intact, allowing adequate estimation of pulmonary artery pressure, in 15 patients (6.2% of all patients with severe TR; idiopathic TR group). Patients with idiopathic TR were older (76 +/- 10 years), with a high frequency of atrial fibrillation (93%), and prominent TV annular dilatation. CONCLUSIONS: After excluding multiple potential causes of TR, severe TR is occasionally idiopathic. Annular dilatation (secondary to aging, atrial fibrillation, or other causes) is the likely mechanism of TR in these patients. PMID- 17400121 TI - Anotomic interaction between the aortic root and the atrial septum: a prospective echocardiographic study. AB - BACKGROUND: We recently demonstrated that patients with platypnea-orthodeoxia syndrome and an enlarged aortic root had a smaller and hypermobile atrial septum (AS) compared with those with a normal aortic root. However, this was a partly retrospective study. METHODS: In all, 72 patients underwent transesophageal echocardiography and cardiac catheterization. The aortic root diameter, AS dimension, AS oscillation amplitude (ASo), and atrial pressure gradient were measured. RESULTS: Significant correlations were found: aortic root diameter and AS dimension (r = -0.5, P < .001), aortic root diameter and ASo (r = +0.3, P = .014), AS dimension and ASo (r = -0.28, P = .02), and ASo and atrial pressure gradient (r = -0.36, P = .003). Nineteen patients presented with patent foramen ovale; those with grade 3 shunting had significantly higher mobility of the AS and larger aortic roots. CONCLUSION: These results confirm that an increasing aortic size affects the AS by decreasing its apparent size and increasing its mobility. In case of a patent foramen ovale, increased AS mobility is associated with greater shunting. PMID- 17400122 TI - Effort angina, normal coronary angiogram, and dynamic left ventricular obstruction. AB - The development of dynamic left ventricular outflow tract (LVOT) obstruction (DLVOTO) during effort in patients without hypertrophic cardiomyopathy has been described only a few times and its clinical significance has not been established. This study was undertaken to determine DLVOTO during exercise in patients with angina and normal coronary arteries, and to analyze the potential mechanism involved. The study included 23 patients (60.7 +/- 7.2 years), with effort angina and normal coronary arteries who underwent exercise echocardiography. DLVOTO during exercise appeared in 7 patients (30.4%), with gradients ranging from 31.1 to 67.2 mm Hg (mean 41.3 +/- 12.6). These patients showed more symptoms (effort angina) during testing than the remaining patients (100% vs 47.3%; P = .014). LVOT size was the only predictive factor of DLVOTO (9.1 +/- 0.5 vs 10.3 +/- 1.2 mm/m(2); P = .017). In patients with angina and no epicardical coronary artery disease DLVOTO is associated with a lower LVOT size. PMID- 17400123 TI - Coronary flow velocity changes in response to hypercapnia: assessment by transthoracic Doppler echocardiography. AB - BACKGROUND: The effects of hypercapnia on coronary arteries in human beings are not known. We used transthoracic Doppler echocardiography to evaluate coronary blood flow velocity (CFV) changes in response to hypercapnia in healthy adults. METHODS: Twenty adults underwent transthoracic Doppler echocardiography of the left anterior descending coronary artery while breathing room air, 40% fraction of inspired oxygen, and 40% fraction of inspired oxygen with carbon dioxide supplemented to end-tidal tensions of +5, +7.5, and +10 mm Hg above baseline. RESULTS: Mean (SD) diastolic peak CFV values for these conditions were 23.1 (9.1), 23.0 (9.0), 25.5 (9.3), 27.9 (11.5), and 31.5 (13.0) cm/s, respectively. Significant overall differences between conditions (P < .001) and progressive levels of hypercapnia (P < or = .01) were observed. CFV increases remained significant after adjusting for increases in cardiac output (P = .038). CONCLUSIONS: CFV increases with hypercapnia. This is the first report of human coronary artery flow responses to hypercapnia. Transthoracic Doppler echocardiography methodology is feasible for measuring CFV and the effects of hypercapnia on the coronary circulation. PMID- 17400124 TI - Guidelines for performing a comprehensive epicardial echocardiography examination: recommendations of the American Society of Echocardiography and the Society of Cardiovascular Anesthesiologists. PMID- 17400125 TI - Cardioembolic stroke in a patient with spindle cell sarcoma of the left atrium. PMID- 17400126 TI - Aortic pseudoaneurysm with fistulization into the pulmonary artery: a cause of refractory heart failure after bentall procedure. AB - We present the case of a patient who developed left-sided congestive heart failure several years after undergoing composite graft surgery for annuloaortic ectasia and aortic regurgitation. Transthoracic echocardiography showed markedly elevated left ventricular filling pressures and severe pulmonary hypertension. However, no underlying pathology or anatomic abnormality to explain the high filling pressures could be identified. On transesophageal echocardiography, a large aortic pseudoaneurysm was demonstrated arising from dehiscence of the distal graft anastomosis to the native aorta. A fistulous communication was noted between the pseudoaneurysm and the right pulmonary artery resulting in a large aortopulmonary shunt. This case illustrates the inherent limitations of transthoracic echocardiography in the detection of complications after composite graft surgery of the aorta. PMID- 17400127 TI - Dual thrombosis of the pulmonary arterial and venous anastomotic sites after single lung transplantation: role of transesophageal echocardiography in diagnosis and management. AB - We present the case of a patient who developed severe respiratory and hemodynamic compromise shortly after single right lung transplantation. Transesophageal echocardiography showed a large intraluminal thrombus at the right pulmonary artery anastomosis resulting in severe obstruction to flow together with thrombosis of the right pulmonary veins extending into the left atrium. After thrombectomy and surgical revision of the vascular anastomoses, the patient made an uneventful recovery. This case illustrates the usefulness of transesophageal echocardiography in the diagnosis and treatment of patients who are hemodynamically unstable after lung transplantation. PMID- 17400128 TI - Isolated noncompaction of the ventricular myocardium: infrequent because of missed diagnosis? AB - Isolated noncompaction of the ventricular myocardium is frequently mistaken for other cardiomyopathies. We report a case of a 49-year-old woman admitted to hospital for heart failure and initially given the diagnosis of apical hypertrophic cardiomyopathy. In this case, myocardial contrast echocardiography and magnetic resonance imaging played a pivotal role in establishing the diagnosis of isolated noncompaction of the ventricular myocardium. PMID- 17400129 TI - Recurrence of takotsubo cardiomyopathy with variant forms of left ventricular dysfunction. AB - Apical ballooning syndrome, or takotsubo cardiomyopathy, is a syndrome characterized by transient left ventricular dysfunction, frequently presenting with electrocardiographic changes and elevated cardiac biomarkers in the absence of obstructive coronary artery disease. We describe a case where repeated emotional stress caused recurrent ventricular dysfunction in varying regions of the left ventricle. PMID- 17400130 TI - A case of platypnea-orthodeoxia syndrome in a patient with a pulmonary arteriovenous fistula and a patent foramen ovale. AB - We describe a case of a rare clinical disorder, platypnea-orthodeoxia syndrome. A 57-year-old man was admitted with progressive dyspnea. While breathing room air, arterial oxygen saturation decreased from 92% in a recumbent position to 83% in an upright position. Transthoracic echocardiography revealed normal left and right ventricular function, but intravenous infusion of air-blood-saline resulted in the appearance of microbubbles in the left side of the heart, indicating the presence of right-to-left shunt. Transesophageal echocardiography demonstrated a small amount of right-to-left shunting through a patent foramen ovale. Further, the left lower pulmonary vein was dilated, and contrast echocardiography revealed drainage of microbubbles mainly from the left lower pulmonary vein. A pulmonary angiogram revealed the presence of a pulmonary arteriovenous fistula. The patient underwent embolization of the arteriovenous fistula with subsequent resolution of dyspnea and hypoxemia. Platypnea-orthodeoxia syndrome is rare, and patients with this syndrome require an extensive workup to determine its cause. PMID- 17400131 TI - Anesthetics and echocardiographic assessment of left ventricular function: lessons learned from invasive analysis of cardiovascular mechanics. PMID- 17400132 TI - Treatment of hypocalcemia in critical illness--part 1. PMID- 17400133 TI - Table for who? PMID- 17400134 TI - Restoration of binocular vision in Guillain-Barre syndrome would improve quality of life. PMID- 17400136 TI - Occurrence of oculomotor dysfunctions in acquired brain injury: a retrospective analysis. AB - BACKGROUND: The purpose of this retrospective study was to determine the frequency of occurrence of oculomotor dysfunctions in a sample of ambulatory outpatients who have acquired brain injury (ABI), either traumatic brain injury (TBI) or cerebrovascular accident (CVA), with associated vision symptoms. METHODS: Medical records of 220 individuals with either TBI (n = 160) or CVA (n = 60) were reviewed retrospectively. This was determined by a computer-based query spanning the years 2000 through 2003, for the frequency of occurrence of oculomotor dysfunctions including accommodation, version, vergence, strabismus, and cranial nerve (CN) palsy. RESULTS: The majority of individuals with either TBI (90%) or CVA (86.7%) manifested an oculomotor dysfunction. Accommodative and vergence deficits were most common in the TBI subgroup, whereas strabismus and CN palsy were most common in the CVA subgroup. The frequency of occurrence of versional deficits was similar in each diagnostic subgroup. CONCLUSION: These new findings should alert the clinician to the higher frequency of occurrence of oculomotor dysfunctions in these populations and the associated therapeutic, rehabilitative, and quality-of-life implications. PMID- 17400137 TI - Topical interferon alpha 2 beta therapy in the management of conjunctival papilloma. AB - BACKGROUND: This article describes the differential diagnosis of conjunctival papillomas and reviews the treatment options with a focus on the new topical interferon alpha 2 beta therapy. CASE: A 45-year-old white man presented with a red right eye of approximately 6 months' duration. The lesion was suspicious and suggestive of malignancy; therefore, the patient was referred for consultation. The lesion was diagnosed as a conjunctival papilloma. The patient was placed on topical interferon alpha 2 beta eye drops, and within 2 weeks he experienced complete lesion regression. CONCLUSION: Interferons, in particular interferon alpha-2b (IFN-alpha2beta), have recently been shown to be successful in treating conjunctival papillomas and conjunctival intraepithelial neoplasia (CIN). Previously, treatment was limited to observation for asymptomatic patients. Larger lesions in symptomatic patients were treated by surgical excision and cryotherapy. Additional treatment modalities included topical antimetabolite treatment with mitomycin-C and 5-fluorouracil. These treatments, although effective, had significant attendant postsurgical complications and toxic ocular adverse reactions. Interferons represent a new successful treatment modality. PMID- 17400138 TI - A case of primary congenital glaucoma: a diagnostic dilemma. AB - BACKGROUND: Primary congenital glaucoma generally presents with a classic clinical triad of photophobia, blepharospasm, and epiphora caused by the corneal changes that occur secondary to increased intraocular pressure (IOP). The condition typically presents bilaterally and is rarely hereditary. Onset is from age 2 months to 2 to 3 years. CASE REPORT: A 2-year, 5-month-old Hispanic boy presented with an enlarged right eye and an intermittent right exotropia, without tearing or photophobia. Examination also found high myopia and an optic nerve cup to-disc ratio larger in the right than the left eye. Referral to a pediatric ophthalmologist was initiated. On the first examination under anesthesia (EUA), the child was diagnosed with unilateral megalocornea with a normal IOP. He did not have any other typical signs and symptoms of primary congenital glaucoma. An EUA 8 months later led to a diagnosis of primary congenital glaucoma based on the new appearance of Haab's striae, further enlargement of the cornea, and an elevated IOP. At this point, medical management was instituted. CONCLUSION: This case shows the importance of recognizing signs of primary congenital glaucoma so that appropriate management can begin as soon as possible to provide the best visual outcome for a child. PMID- 17400139 TI - Solitary idiopathic choroiditis. AB - Solitary idiopathic choroiditis (SIC) is often confused with intraocular masses and inflammatory conditions and has only recently been identified as a distinct clinical condition. Jerry Shields, M.D., was the first to put a name on this condition. The clinical findings and features that distinguish it from other conditions are discussed in this report. A comprehensive examination with supplemental testing such as fluoresceine angiography, B scan, and extensive laboratory workup is necessary to rule out the differentials of SIC. These differentials include inflammatory conditions and retinal and choroidal tumors. SIC has both an active and an inactive phase. The treatment for the condition is controversial. Our patient presented with a suspicious intraocular mass that was diagnosed as SIC after an extensive ocular examination and laboratory workup was completed. PMID- 17400140 TI - An assessment of regional differences in corneal thickness in normal human eyes, using the Orbscan II or ultrasound pachymetry. AB - BACKGROUND: In recent years, there has been increasing interest in the characteristics of the peripheral cornea close to the limbus, in both tonometry measures and refractive surgery, but there is relatively little information on these characteristics as provided by modern day pachymetry instruments such as the Orbscan (Bausch & Lomb, Rochester, New York). The current study was therefore undertaken to assess the corneal thickness profile along the horizontal meridian by this scanning slit light method, comparing the data with that obtained with an ultrasound pachymeter. METHODS: Noncontact specular microscopy was first performed on 17 adults (aged between 20 and 64 years) to check that the corneas were normal. Then, 3 assessments of the corneal thickness profile across the horizontal meridian were taken using the Orbscan II, and both the regional map data (7-mm-diameter measurement ring, 1-mm sample zones) and the point data from the numerical pachymetry output were used to extract data at specific locations nominally 0.5 mm apart. Ultrasound pachymetry (under topical anesthesia with benoxinate 0.4%) was then used to obtain thickness values at central, mid peripheral (2.75 mm), and peripheral locations close to the limbus (4.5 mm). RESULTS: Specular microscopy yielded mean thickness of 0.529 +/- 0.032 mm, whereas single-point Orbscan readings at the geometric center of the cornea averaged 0.579 +/- 0.037 mm. Orbscan readings around the 7-mm-diameter measurement zone along the horizontal meridian averaged 0.681 +/- 0.034 mm (i.e., were 0.101 mm or 17.6% greater; P < 0.001). Mid-peripheral readings taken from the numerical maps at 2.5 to 3.0 mm averaged 0.645 mm (or 11% higher than central point readings), whereas peripheral readings between 4.0 and 4.5 mm averaged 0.727 mm (i.e., 26% higher than central point values). In marked contrast, ultrasound readings in the mid-periphery (2.75 mm) averaged just 0.553 mm (or 5.5% greater than the central corneal thickness [CCT]) and just 0.612 mm (i.e., 16.6% higher) in the periphery (4.5 mm). CCT profiles generated from the Orbscan numerical output across the horizontal meridian showed a predictable progressive increase in thickness from the center to the 4.5-mm location on both the temporal and nasal side. The mean differences between the Orbscan II and ultrasound pachymetry measures were thus not constant across the cornea. Without correction, these differences were close to 0.05 mm at the center but more than 0.100 mm at the peripheral sites, and proportional differences persisted after application of the default acoustic factor of 0.92 for the Orbscan readings. CONCLUSIONS: Orbscan II and ultrasound pachymetry measures generate a rather different profile for corneal thickness. The data from the 2 techniques should be considered as reporting different characteristics, rather than attempts being made to align Orbscan measurements to those of the ultrasound method. A single acoustic correction factor cannot be logically applied to all corneal thickness measures made with an Orbscan II. PMID- 17400141 TI - Journeying with Peter and Sam: reflections on a healing attitude. AB - Reflection enables the practitioner to learn through everyday experiences towards realising desirable practice however it is articulated. In this paper I share my story in journeying with Peter and Sam in a hospice setting with the intention to ease suffering. In considering what is significant within this story, I pay particular attention to the idea of a healing attitude for deeper reflection in order to develop my appreciation of the nature and significance of a healing attitude. PMID- 17400142 TI - Qigong reduces stress in computer operators. AB - Chinese research indicates that the Qigong method reduces psychosomatic and physical symptoms through an effect on the sympathetic nervous system. OBJECTIVES: The aim was to investigate the effects of Qigong on stress among computer operators. DESIGN: Ten women were included in a Qigong group and an equal number in a control group. Heart rate, blood pressure, and finger temperature were measured at the beginning and at the end of the working day during 5 weeks. twenty four-hours urine samples were collected in the first and last weeks to measure catecholamine excretion in urine. Participants kept a daily record of psychological measures of strain and weekly measures of stress levels. RESULTS AND CONCLUSIONS: Qigong reduced noradrenaline excretion in urine (p<0.05), and influenced the heart rate and temperature, indicating reduced activity of the sympathetic nervous system. Moreover, Qigong reduced low-back symptoms (p<0.05). In conclusion, Qigong exercise may reduce stress at computerised work. PMID- 17400143 TI - The effects of prayer, relaxation technique during general anesthesia on recovery outcomes following cardiac surgery. AB - During general anesthesia the possibility of subconscious perception of intraoperative events is a controversial subject. Some studies found that positive verbal suggestions, or music improved intraoperative relaxation and postoperative recovery. The aim of the current study was to evaluate the effect of prayer and relaxation technique applied while patients are under general anesthesia for open-heart surgery. A randomized, controlled, double-blind trial study included 78 patients who underwent cardiac surgery. During the surgery the patients used a headphone connected to a CD player. They were randomly divided into three groups. One group listened to prayer during the surgery, the other listened to relaxation technique and one, placebo. There was only one significant finding: the prayer group is less likely to believe that prayer would assist conventional medical treatments. Although not statistically significant, we discussed the length of stay (LOS) after surgery and the incidence of sternal wound infection. PMID- 17400144 TI - Evaluation of a multi-component approach to cognitive-behavioral therapy (CBT) using guided visualizations, cranial electrotherapy stimulation, and vibroacoustic sound. AB - This pilot study examines the use of guided visualizations that incorporate both cognitive and behavioral techniques with vibroacoustic therapy and cranial electrotherapy stimulation to form a multi-component therapeutic approach. This multi-component approach to cognitive-behavioral therapy (CBT) was used to treat patients presenting with a range of symptoms including anxiety, depression, and relationship difficulties. Clients completed a pre- and post-session symptom severity scale and CBT skills practice survey. The program consisted of 16 guided visualizations incorporating CBT techniques that were accompanied by vibroacoustic therapy and cranial electrotherapy stimulation. Significant reduction in symptom severity was observed in pre- and post-session scores for anxiety symptoms, relationship difficulties, and depressive symptoms. The majority of the clients (88%) reported use of CBT techniques learned in the guided visualizations at least once per week outside of the sessions. PMID- 17400145 TI - First impressions in complementary practice: the importance of environment, dress and address to the therapeutic relationship. AB - The aim of the study was to explore patients' views of the importance of environmental and social factors within a complementary and alternative medicine (CAM) practice. A cross-sectional patient survey was conducted in 37 CAM practices across nine geographical areas of the UK using a structured questionnaire. The 219 responders (response rate 59%) were 65% female. The most common treatment modalities were osteopathy and acupuncture. Patients rated telephone manner, practitioner appearance and explanatory literature as very important factors in shaping first impressions and influencing their confidence in the complementary practice. Patients' preferences for practitioner dress code were highly variable. Overall 65% preferred a white coat to casual dress. A multivariate analysis revealed that this preference was statistically significantly greater in two geographical regions and in patients over 70 years old. It was not influenced by gender or treatment modality. Further research is recommended in order to understand how the complex verbal and non-verbal communication implicit in the CAM setting may affect the therapeutic relationship. PMID- 17400146 TI - Enhancing mental health services to bone marrow transplant recipients through a mindfulness-based therapeutic intervention. AB - Complementary and alternative therapies are gaining recognition in the treatment of many disease states. The importance of treating psychological and emotional problems associated with bone marrow transplant has been substantiated by research evidence. This feasibility study tested a mindfulness-based therapeutic intervention to treat such problems in this context. Pretests and post-tests were administered to patients (n=24) undergoing bone marrow transplant. Results indicate that the mindfulness-based therapeutic intervention has the potential to be an effective therapy for bone marrow transplant recipients. PMID- 17400147 TI - Dissociation reduction in body therapy during sexual abuse recovery. AB - The study purpose was to examine dissociation in body therapy for women receiving psychotherapy for childhood sexual abuse. An initial intervention study provided an opportunity to examine dissociation; the sample of 24 women received eight, 1 h body therapy sessions. The Dissociative Experiences Scale served as the predictor variable, and the outcome measures reflected psychological and physical health, and body connection. Repeated measures analysis of variance was used to examine dissociation reduction across time. Pearson correlations were used to describe associations between the relative change in dissociation and outcomes. The results demonstrated that the greatest change was the reduction of dissociation; there was an incremental effect across time and a strong association between change in dissociation and health outcomes. High dissociation at baseline (moderate levels) predicted positive outcomes. The results demonstrated the importance of moderate dissociation as an indicator of distress, and the central role of dissociation reduction in health and healing. PMID- 17400148 TI - Midwifery managers' views about the use of complementary therapies in the maternity services. AB - The burgeoning interest in complementary therapies (CTs) in the general population over the last decade has created a demand for CTs to be made available within the NHS. There are some excellent examples of midwives who have introduced CTs into clinical practice and who are providing an enhanced service to women as a result [Budd S. Moxibustion for breech presentation. Complement Therap Nurs Midwifery 2000; 6(4): 176-9; Tiran D. Complementary strategies in antenatal care. Complement Therap Nurs Midwifery 2001; 7: 19-24; Ager C. A complementary therapy clinic, making it work. RCM Midwives J 2002; 5(6): 198-200; Burns E, Blamey C, Ersser S, Lloyd AJ, Barnetsson L. The use of aromatherapy in intrapartum midwifery practice: an observational study. Oxford: OCHRAD; 1999]. Overall, however, service provision remains patchy and ad hoc with little evidence of a robust integration into the maternity services. This article presents the qualitative findings from a national survey of the heads of maternity services in England. They were asked to indicate their views and perceptions about the benefits, promoters and constrainers in relation to CT integration within the maternity services. Our findings show that overall, views are positive, with increasing consumer satisfaction, promotion of normal childbirth and a reduction in medical intervention being seen as the main benefits. PMID- 17400151 TI - Early management of the traumatized airway. AB - Penetrating face and neck trauma is usually obvious, but blunt trauma mandates high index of suspicion to recognize its existence. Comprehensive understanding of the injury is mandatory to plan the best timing and method to secure the airway. PMID- 17400152 TI - The critically ill injured patient. AB - Patients admitted to the ICU after severe trauma require frequent procedures in the operating room, particularly in cases where a damage control strategy is used. The ventilatory management of these patients in the operating room can be particularly challenging. These patients often have severely impaired respiratory mechanics because of acute lung injury and abdominal compartment syndrome. Consequently, the pressure and flow generation capabilities of standard anesthesia ventilators may be inadequate to support ventilation and gas exchange. This article presents the problems that may be encountered in patients who have severe abdominal and lung injuries, and the current management concepts used in caring for these patients in the critical care setting, to provide guidelines for the anesthetist faced with these patients in the operating room. PMID- 17400153 TI - Current concepts in hemorrhagic shock. AB - Treatment of hemorrhagic shock continues to evolve. Surgically, the concept of "damage control" has become the standard, with shorter operations focused on the control of life-threatening injuries. For the anesthesiologist, practice has changed, with a better understanding of the pathophysiology of shock, new techniques such as deliberate hypotension and perfusion-focused therapy, improved management of transfusion, and the advent of systemic hemostatic drug therapy. In this article, the author reviews each of these topics, and presents an integrated recommendation for early and late management of the patient in hemorrhagic shock. PMID- 17400154 TI - Nonsurgical treatment of major bleeding. AB - Novel nonsurgical approaches to bleeding control offer hope for improved management of the critical trauma-related coagulopathy and diffuse bleeding that often typify major trauma and pose challenges to surgeons and anesthetists. Although surgical treatment is the cornerstone of bleeding control, in selected patients angiographic embolization is increasingly used early in patient care to successfully manage arterial bleeding attributable to blunt solid organ injury or posterior pelvic ring disruption. Coagulopathic derangements in trauma occur early and require avoidance or correction of acidosis and hypothermia. If bleeding cannot be stopped by these measures, adjunctive use of fibrinogen or recombinant activated factor VII (rFVIIa) have the potential to correct systemic coagulopathy associated with massive blood loss and its management. PMID- 17400155 TI - Trauma and aggressive homeostasis management. AB - Three important issues concerning homeostasis in the acute care of trauma patients that are related directly to the stress response are hyperglycemia, lactic acidosis, and hypothermia. Recently, there has been a resurgence of interest in investigating the effects of aggressive thermal and glucose concentration and volume resuscitation on outcomes in critically ill and trauma patients. Significant reason exists to question the "conventional wisdom" relating to current approaches to restoring homeostasis in this patient population. PMID- 17400156 TI - Training and assessment of trauma management: the role of simulation-based medical education. AB - Simulation-based medical education (SBME) offers a safe and "mistake-forgiving" environment to teach and train medical professionals. The diverse range of medical simulation modalities enables trainees to acquire and practice an array of tasks and skills. SBME offers the field of trauma training multiple opportunities to enhance the effectiveness of the education provided in this challenging domain. In this article, the authors describe the possible roles of simulated patients, skills trainers, computerized patient simulators, and web based teaching in trauma training, and describe some practical aspects of using simulation for trauma training. PMID- 17400157 TI - Geriatric trauma: special considerations in the anesthetic management of the injured elderly patient. AB - Modern society is characterized as having an ever enlarging population of older adults. There are more elderly patients, and the average age of this group is increasing. The anesthetic management of surgery for the elderly trauma victim is more complicated than in younger adults. Evaluation of the physiologic status of the geriatric patient should take into account the variability of the changes associated with advancing age. Care of the injured elderly patient requires thorough preoperative assessment and planning and the involvement of a multidisciplinary clinical team knowledgeable about and interested in the management of the elderly surgical patient. PMID- 17400158 TI - Management of the obese trauma patient. AB - Obese persons are more likely to be involved in vehicle accidents, probably because of the presence of sleep apnea. They are more likely to suffer chest, pelvis, and extremity fractures. Mildly overweight persons are less prone to intra-abdominal injury because of the protective effect of the abdominal fat, known as the cushion effect. Obese trauma patients are far more likely to develop in-hospital complications, especially pulmonary, renal, and thromboembolic complications. The BMI is an independent risk factor for morbidity and mortality after trauma. Because only limited data exist about the right clinical approach to obese trauma patients, it is necessary to rely on general knowledge about treating obese patients in the ICU. More research is needed to improve the treatment of obese trauma patients. PMID- 17400159 TI - Regional anesthesia in trauma patients. AB - Medical and surgical treatment of the trauma patient has evolved in the last decade. Treatment of pain from multiple fractures or injured organs and surgical anesthesia with regional anesthesia techniques have been used to reduce post traumatic stress disorder and reduce the adverse effects of general anesthesia. Neuraxial blocks and peripheral nerve block techniques should be practiced by trained emergency and operatory room staff. This article reviews recent publications related to the role of regional anesthesia in trauma patients in the prehospital, emergency, and operatory room settings. It also describes indications, limitations, and practical aspects of regional anesthesia. PMID- 17400160 TI - Initial trauma management in advanced pregnancy. AB - The principles enshrined in existing trauma resuscitation protocols for treating nonpregnant trauma victims should also be applied to the pregnant patient. In addition, left tilt of the pregnant patient (or the back board) and supplement oxygen are mandatory. The patient should be treated by a multidisciplinary team, preferably in a trauma center. Early intubation is recommended, but should be performed, where possible, by an experienced physician. The physician should be aware of the different physiologic and laboratory values in normal pregnancy. Fetal monitoring is important to assess both fetal and maternal welfare. Imaging examinations, where indicated, should not be delayed. Even minor maternal trauma, especially if caused by interpersonal violence, might cause fetal loss. PMID- 17400161 TI - Battlefield anesthesia: advances in patient care and pain management. AB - Expeditionary maneuver warfare and the asymmetric battlefield have forced changes in the traditional methods with which we deliver anesthesia and surgery to the wounded. Although in many ways similar to how we have operated on the wounded for the past half century, new advances in diagnostic and therapeutic modalities and doctrinal shifts have changed the face of the battlefield hospital. In this article, the authors discuss these changes in regard to anesthetic care for surgical and pain management for wounded airmen, sailors, soldiers, and marines. PMID- 17400162 TI - Mechanisms of injury by explosive devices. AB - Explosive devices cause injury by four mechanisms, of which primary blast injury is the least familiar to most non-military clinicians. The pathophysiology of the various mechanisms of injury is described, and the implications for translating a knowledge of mechanism of injury to clinical management is discussed. PMID- 17400163 TI - Disaster preparedness, triage, and surge capacity for hospital definitive care areas: optimizing outcomes when demands exceed resources. AB - Disaster planning must anticipate how demands imposed by a disaster reconcile with the capacity of the treating facility. Resources must be organized before an event so that they are optimally used to treat as many victims as possible, as well as to avoid overwhelming available resources. PMID- 17400164 TI - Multiple casualty incidents: the prehospital role of the anesthesiologist in Europe. AB - The recent increase in incidents involving mass casualties has emphasized the need for a planned and coordinated prehospital emergency medical response, with medical teams on-site to provide advanced trauma life support. The special skills of the anesthesiologist make his/her contribution to prehospital emergency care particularly valuable. The United Kingdom's emergency medical services system is operated paramedically like that in the United States, and is based on rapid evacuation of casualties to hospital emergency medical facilities. In contrast, the French approach is based on the use of its emergency care system SAMU, where both structured dispatching and on-site medical care is provided by physicians, including anesthesiologists. In this article, the lessons learned from multiple casualty incidents in Europe during the past 2 decades are considered from the standpoint of the anesthesiologist. PMID- 17400165 TI - Nonconventional terror--the anesthesiologist's role in a nerve agent event. AB - The structure and biologic action of nerve agents is similar to organophosphates, commonly used as insecticides. Acetylcholine accumulation and binding to the cholinergic receptor site stimulates the affected organs producing a predictable set of clinical symptoms. Treatment of the affected patients will include decontamination, respiratory and hemodynamic support, as well as specific antidotes. The multiple casualties that may be expected present additional logistical and organizational problems. The specific skills of anesthesiologists will make them invaluable members of the care team in such a chemical mass casualty event. PMID- 17400166 TI - Reverse micelles: inert nano-reactors or physico-chemically active guides of the capped reactions. AB - Reverse micelles present self-assembled multi-molecular entities formed within specific compositional ranges of water-in-oil microemulsions. The structure of a reverse micelle is typically represented as nano-sized droplet of a polar liquid phase, capped by a monolayer of surfactant molecules, and uniformly distributed within a non-polar, oil phase. Although their role in serving as primitive membranes for encapsulation of primordial self-replicating chemical cycles that anticipated the very origins of life has been proposed, their first application for 'parent(hesis)ing' chemical reactions with an aim to produce 'templated' 2D arrays of nanoparticles dates back to only 25 years ago. Reverse micelles have since then been depicted as passive nano-reactors that via their shapes template the growing crystalline nuclei into narrowly dispersed or even perfectly uniform nano-sized particles. Despite this, numerous examples can be supported, where from deviations from the simple unilateral correlations between size and shape distribution of reverse micelles and the particles formed within may be reasonably implied. A rather richer, dynamical role of reverse micelles, with potential significance in the research and design of complex, self-assembly synthesis pathways, as well as possible adoption of their application as an aspect of biomimetic approach, is suggested herein. PMID- 17400167 TI - Characterization of cap-shaped silver particles for surface-enhanced fluorescence effects. AB - Surface-enhanced fluorescence has potentially many desirable properties as an analytical method for medical diagnostics, but the effect observed so far is rather modest and only in conjunction with fluorophores with low quantum yields. Coupled with the fact that preparation of suitable surfaces at low costs has been difficult, this has limited its utilities. Here we report a novel method for forming uniform and reproducible surfaces with respectable enhancement ratios even for high-quantum-yield fluorophores. Formation of dense surface-adsorbed latex spheres on a flat surface via partial aggregation, followed by evaporation of silver, results in a film consisting of cap-shaped silver particles at high densities. Binding of fluorescence biomolecules, either through physisorption or antigen-antibody reaction, was performed, and enhancements close to 50 have been observed with fluorophores such as R-phycoerythrin and Alexa 546-labeled, bovine serum albumin, both of which have quantum yields around 0.8. We attribute this to the unique shape of the silver particle and the presence of abundant gaps among adjacent particles at high densities. The effectiveness of the new surface is also demonstrated with IL-6 sandwich assays. PMID- 17400168 TI - Functional identification of novel activities: activity-based selection of proteins from complete proteomes. AB - The identification of proteins with desired activities, especially from complex samples such as plasma and whole blood, is a continual challenge. We have developed a technology platform called Functional Identification of Novel Activities (FIoNA) to discover desired protein activities from complex biological samples. FIoNA uses immobilized libraries of combinatorial peptide ligands to purify and concentrate essentially all of the components of a complex mixture on ligands synthesized on individual beads. No depletion or prefractionation of the starting material is performed before it is incubated with the library, and no a priori knowledge of the active protein or of the ligand to which it binds is required. Instead, the protein-loaded beads are individually evaluated en masse in disease- relevant assays to identify proteins possessing a desired function. Beads associated with the activity are selected, and the ligand is sequenced and resynthesized in bulk on the original backbone for purification and characterization of the active component. Here we illustrate the use of FIoNA in a cell proliferation assay to detect a growth factor present in conditioned cell medium at nanogram/milliliter concentrations. We also have selected beads associated with hydrolysis of nerve agent analogs in assays performed in 100,000 well microtiter plates. PMID- 17400169 TI - Using cadmium telluride quantum dots as a proton flux sensor and applying to detect H9 avian influenza virus. AB - Semiconductor nanocrystals, often known as quantum dots, have been used extensively for a wide range of applications in bioimaging and biosensing. In this article, we report that the pH-sensitive cadmium telluride (CdTe) quantum dots (QDs) were used as a proton sensor to detect proton flux that was driven by ATP synthesis in chromatophores. To confirm that these QD-labeled chromatophores were responding to proton flux pumping driven by ATP synthesis, N,N' dicyclohexylcarbodiimide (DCCD) was used as an inhibitor of ATPase activity. Furthermore, we applied the QD-labeled chromatophores as a virus detector to detect the H9 avian influenza virus based on antibody-antigen reaction. The results showed that this QD virus detector could be a new virus-detecting device. PMID- 17400170 TI - Application of RNase in the purification of RNA-binding proteins. PMID- 17400171 TI - A high-throughput screening platform for inhibitors of the riboflavin biosynthesis pathway. AB - 3,4-Dihydroxy-2-butanone 4-phosphate synthase, 6,7-dimethyl-8-ribityllumazine synthase, and riboflavin synthase of the riboflavin biosynthetic pathway are potential targets for novel antiinfective drugs. This article describes a platform for high-throughput screening for inhibitors of these enzymes. The assays can be monitored photometrically and have been shown to be robust, as indicated by Z factors 0.87. A (13)C NMR assay for hit verification of 3,4 dihydroxy-2-butanone 4-phosphate synthase inhibitors is also reported. PMID- 17400172 TI - Development of an analytical HPLC system for the specific detection of 2 isopropylmalic acid in yeast growth medium. PMID- 17400173 TI - Observed finger behaviour during computer mouse use. AB - Two-button computer mouse users may exhibit sustained, static finger lifting behaviours to prevent inadvertent activations by avoiding finger pressure on the buttons, which leads to prolonged, static finger extensor muscle loading. One hundred graduate students were observed during normal computer work in a university computer facility to qualify and quantify the prevalence of lifted finger behaviours and extended finger postures, as well as wrist/forearm and grip behaviour, during specific mouse activities. The highest prevalences observed were 48% of the students lifted their middle finger during mouse drag activities, and 23% extended their middle finger while moving the mouse. In addition, 98% of the students rested their wrist and forearm (77%) or wrist only (21%) on the workstation surface, and 97% had an extended wrist posture (15 degrees -30 degrees ) when using the mouse. Potential applications of these findings include future computer input device designs to reduce finger lifting behaviour and exposures to risk factors of hand/forearm musculoskeletal pain. PMID- 17400174 TI - Functional expression and characterization of cytochrome P450 52A21 from Candida albicans. AB - Candida albicans contains 10 putative cytochrome P450 (CYP) genes coding for enzymes that appear to play important roles in fungal survival and virulence. Here, we report the characterization of CYP52A21, a putative alkane/fatty acid hydroxylase. The recombinant CYP52A21 protein containing a 6x(His)-tag was expressed in Escherichia coli and was purified. The purified protein, reconstituted with rat NADPH-cytochrome P450 reductase, omega-hydroxylated dodecanoic acid to give 12-hydroxydodecanoic acid, but to a lesser extent also catalyzed (omega-1)-hydroxylation to give 11-hydroxydodecanoic acid. When 12,12,12-d(3)-dodecanoic acid was used as the substrate, there was a major shift in the oxidation from the omega- to the (omega-1)-hydroxylated product. The regioselectivity of fatty acid hydroxylation was examined with the 12-iodo-, 12 bromo-, and 12-chlorododecanoic acids. Although all three 12-halododecanoic acids bound to CYP52A21 with similar affinities, the production of 12-oxododecanoic acid decreased as the size of the terminal halide increased. The regioselectivity of CYP52A21 fatty acid oxidation is thus consistent with presentation of the terminal end of the fatty acid chain for oxidation via a narrow channel that limits access to other atoms of the fatty acid chain. This constricted access, in contrast to that proposed for the CYP4A family of enzymes, does not involve covalent binding of the heme to the protein. PMID- 17400175 TI - Leptin attenuates gene expression for renal 25-hydroxyvitamin D3-1alpha hydroxylase in mice via the long form of the leptin receptor. AB - Leptin, the ob gene product secreted by adipocytes, controls overall energy balance. We previously showed that leptin administration to leptin-deficient obese (ob/ob) mice suppressed mRNA expression and activity of renal 25 hydroxyvitamin D(3)-1alpha-hydroxylase (CYP27B1). In leptin receptor-deficient (db/db) mice, we presently examined whether leptin affects 1alpha-hydroxylase expression in renal tubules through the active form of the leptin receptor (ObRb). Elevated serum concentrations of calcium and 1,25-dihydroxyvitamin D(3) [1,25-(OH)(2)D(3)] in untreated ob/ob mice showed sharp reduction with leptin administration (4 mg/kg, i.p. every 12h for 2 days); no such reduction of elevation occurred in db/db mice. ObRb mRNA was expressed in kidney, brain, fat, lung, and bone in wild-type and ob/ob mice, but not db/db mice. The ob/ob and db/db mice showed large increases in renal 1alpha-hydroxylase mRNA expression and activity. Leptin administration (4 mg/kg) completely abrogated these increases in ob/ob but not db/db mice. Renal 25-hydroxyvitamin D(3)-24-hydroxylase (CYP24) mRNA synthesis also was greatly elevated in ob/ob and db/db mice; excesses decreased significantly with leptin administration in ob/ob mice, but increased in db/db mice. Renal tubular cells in primary culture expressed mRNAs including proximal tubules markers (1alpha-hydroxylase and megalin), parathyroid hormone receptor, and vitamin D receptor. Calcitonin receptor mRNA, synthesized mainly in distal tubules, was scant, indicating that most cultured cells were from proximal tubules. Cells did not express ObRb mRNA. Forskolin exposure at 10(-6)M for 3 or 6h significantly increased 1alpha-hydroxylase mRNA. Leptin at 10(-6)M did not change mRNA expression in either presence or absence of forskolin. Accordingly, leptin attenuates renal 1alpha-hydroxylase gene expression through ObRb. Furthermore, leptin appears to act indirectly on renal proximal tubules to regulate 1alpha-hydroxylase gene expression. PMID- 17400176 TI - A transmembrane osteoclastic protein-tyrosine phosphatase regulates osteoclast activity in part by promoting osteoclast survival through c-Src-dependent activation of NFkappaB and JNK2. AB - This study evaluated the effects of overexpression of wild-type (WT) or phosphatase-deficient (PD) mutant of an osteoclastic protein-tyrosine phosphatase (PTP-oc) in RAW/C4 cells. Osteoclast-like cells derived from WT-PTP-oc overexpressing clones increased, while those derived from PD-PTP-oc expressing clones decreased, their resorption activity. WT-PTP-oc clones had lower apoptosis, lower caspase 3/7 activity, reduced c-Src tyr-527 phosphorylation (PY527) and IkappaBalpha cellular levels, and increased NFkappaB activation and JNK phosphorylation. Overexpression of PD-PTP-oc or PTP-oc siRNA treatment increased apoptosis, caspase 3/7 activity, PY527 and IkappaBalpha levels, and decreased NFkappaB and JNK2 activation. Inhibition of the c-Src kinase blocked the PTP-oc-mediated NFkappaB and JNK2 activation. Blocking the NFkappaB activation had no effect on the JNK2 activation. Inhibiting the NFkappaB and/or JNK2 pathway prevented the PTP-oc-mediated reduction in apoptosis. In conclusion, PTP-oc activates osteoclast activity in part by promoting osteoclast survival through the PTP-oc-mediated c-Src-dependent activation of NFkappaB and JNK2. PMID- 17400177 TI - Zinc as an anti-tumor agent in prostate cancer and in other cancers. AB - Human prostate glandular epithelial cells have the unique capability of accumulating high levels of zinc. This is essential to inhibit m-aconitase activity so that citrate can accumulate for secretion into prostatic fluid, which is a major function of the prostate gland. As a result, the Krebs cycle is truncated with the consequence of the lost ATP production that would result from citrate oxidation. The cellular accumulation of zinc also inhibits mitochondrial terminal oxidation and respiration. In addition to these metabolic effects, zinc accumulation exhibits anti-proliferative effects via its induction of mitochondrial apoptogenesis. Zinc accumulation also inhibits the invasive/migration activities in malignant prostate cells. The anti-proliferative effects and the effects on invasion and migration occur through zinc activation of specific intracellular signaling pathways. Consequently, these effects impose anti-tumor actions by zinc. The ability of prostate cells to accumulate zinc is due to the expression and activity of the zinc uptake transporter, ZIP1. To avoid the anti-tumor effects of zinc, in prostate cancer the malignant prostate cells exhibit a silencing of ZIP1 gene expression accompanied by a depletion of cellular zinc. Therefore we regard ZIP1 as a tumor suppressor gene in prostate cancer. In addition to prostate cells, similar tumor suppressor effects of zinc have been identified in several other types of tumors. PMID- 17400178 TI - Site-directed mutagenesis on the heme axial-ligands of cytochrome b559 in photosystem II by using cyanobacteria Synechocystis PCC 6803. AB - Cytochrome (cyt) b559 has been proposed to play an important role in the cyclic electron flow processes that protect photosystem II (PSII) from light-induced damage during photoinhibitory conditions. However, the exact role(s) of cyt b559 in the cyclic electron transfer pathway(s) in PSII remains unclear. To study the exact role(s) of cyt b559, we have constructed a series of site-directed mutants, each carrying a single amino acid substitution of one of the heme axial-ligands, in the cyanobacterium Synechocystis sp. PCC6803. In these mutants, His-22 of the alpha or the beta subunit of cyt b559 was replaced with either Met, Glu, Tyr, Lys, Arg, Cys or Gln. On the basis of oxygen-evolution and chlorophyll a fluorescence measurements, we found that, among all mutants that were constructed, only the H22Kalpha mutant grew photoautotrophically, and accumulated stable PSII reaction centers ( approximately 81% compared to wild-type cells). In addition, we isolated one pseudorevertant of the H22Ybeta mutant that regained the ability to grow photoautotrophically and to assemble stable PSII reaction centers ( approximately 79% compared to wild-type cells). On the basis of 77 K fluorescence emission measurements, we found that energy transfer from the phycobilisomes to PSII reaction centers was uncoupled in those cyt b559 mutants that assembled little or no stable PSII. Furthermore, on the basis of immunoblot analyses, we found that in thylakoid membranes of cyt b559 mutants that assembled little or no PSII, the amounts of the D1, D2, cyt b559alpha and beta polypeptides were very low or undetectable but their CP47 and PsaC polypeptides were accumulated to the wild-type level. We also found that the amounts of cyt b559beta polypeptide were significantly increased (larger than two folds) in thylakoid membranes of cyt b559 H22YbetaPS+ mutant cells. We suspected that the increase in the amounts of cyt b559 H22YbetaPS+ mutant polypeptides in thylakoid membranes might facilitate the assembly of functional PSII in cyt b559 H22YbetaPS+ mutant cells. Moreover, we found that isolated His-tagged PSII particles from H22Kalpha mutant cells gave rise to redox-induced optical absorption difference spectra of cyt b559. Therefore, our results concluded that significant fractions of H22Kalpha mutant PSII particles retained the heme of cyt b559. Finally, this work is the first report of cyt b559 mutants having substitutions of an axial heme-ligands that retain the ability to grow photoautotrophically and to assemble stable PSII reaction centers. These two cyt b559 mutants (H22Kalpha and H22YbetaPS+) and their PSII reaction centers will be very suitable for further biophysical and biochemical studies of the functional role(s) of cyt b559 in PSII. PMID- 17400179 TI - Two reaction pathways for transformation of high potential cytochrome b559 of PS II into the intermediate potential form. AB - This study describes an analysis of different treatments that influence the relative content and the midpoint potential of HP Cyt b559 in PS II membrane fragments from higher plants. Two basically different types of irreversible modification effects are distinguished: the HP form of Cyt b559 is either predominantly affected when the heme group is oxidized ("O-type" effects) or when it is reduced ("R-type" effects). Transformation of HP Cyt b559 to lower potential redox forms (IP and LP forms) by the "O-type" mechanism is induced by high pH and detergent treatments. In this case the effects consist of a gradual decrease in the relative content of HP Cyt b559 while its midpoint potential remains unaffected. Transformation of HP Cyt b559 via an "R-type" mechanism is caused by a number of exogenous compounds denoted L: herbicides, ADRY reagents and tetraphenylboron. These compounds are postulated to bind to the PS II complex at a quinone binding site designated as Q(C) which interacts with Cyt b559 and is clearly not the Q(B) site. Binding of compounds L to the Q(C) site when HP Cyt b559 is oxidized gives rise to a gradual decrease in the E(m) of HP Cyt b559 with increasing concentration of L (up to 10 K(ox)(L) values) while the relative content of HP Cyt b559 is unaffected. Higher concentrations of compounds L required for their binding to Q(C) site when HP Cyt b559 is reduced (described by K(red)(L)) induce a conversion of HP Cyt b559 to lower potential redox forms ("R type" transformation). Two reaction pathways for transitions of Cyt b559 between the different protein conformations that are responsible for the HP and IP/LP redox forms are proposed and new insights into the functional regulation of Cyt b559 via the Q(C) site are discussed. PMID- 17400180 TI - Influence of poly(ethylene glycol) grafting density and polymer length on liposomes: relating plasma circulation lifetimes to protein binding. AB - The incorporation of poly(ethylene glycol) (PEG)-conjugated lipids in lipid-based carriers substantially prolongs the circulation lifetime of liposomes. However, the mechanism(s) by which PEG-lipids achieve this have not been fully elucidated. It is believed that PEG-lipids mediate steric stabilization, ultimately reducing surface-surface interactions including the aggregation of liposomes and/or adsorption of plasma proteins. The purpose of the studies described here was to compare the effects of PEG-lipid incorporation in liposomes on protein binding, liposome-liposome aggregation and pharmacokinetics in mice. Cholesterol-free liposomes were chosen because of their increasing importance as liposomal delivery systems and their marked sensitivity to protein binding and aggregation. Specifically, liposomes containing various molecular weight PEG-lipids at a variety of molar proportions were analyzed for in vivo clearance, aggregation state (size exclusion chromatography, quasi-elastic light scattering, cryo transmission and freeze fracture electron microscopy) as well as in vitro and in vivo protein binding. The results indicated that as little as 0.5 mol% of 1,2 distearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE) modified with PEG having a mean molecular weight of 2000 (DSPE-PEG(2000)) substantially increased plasma circulation longevity of liposomes prepared of 1,2-distearoyl-sn-glycero-3 phosphatidylcholine (DSPC). Optimal plasma circulation lifetimes could be achieved with 2 mol% DSPE-PEG(2000). At this proportion of DSPE-PEG(2000), the aggregation of DSPC-based liposomes was completely precluded. However, the total protein adsorption and the protein profile was not influenced by the level of DSPE-PEG(2000) in the membrane. These studies suggest that PEG-lipids reduce the in vivo clearance of cholesterol-free liposomal formulations primarily by inhibition of surface interactions, particularly liposome-liposome aggregation. PMID- 17400181 TI - Selectivity signatures of three isoforms of recombinant T-type Ca2+ channels. AB - Voltage-gated Ca(2+) channels (VGCCs) are recognized for their superb ability for the preferred passage of Ca(2+) over any other more abundant cation present in the physiological saline. Most of our knowledge about the mechanisms of selective Ca(2+) permeation through VGCCs was derived from the studies on native and recombinant L-type representatives. However, the specifics of the selectivity and permeation of known recombinant T-type Ca(2+)-channel alpha1 subunits, Ca(v)3.1, Ca(v)3.2 and Ca(v)3.3, are still poorly defined. In the present study we provide comparative analysis of the selectivity and permeation Ca(v)3.1, Ca(v)3.2, and Ca(v)3.3 functionally expressed in Xenopus oocytes. Our data show that all Ca(v)3 channels select Ca(2+) over Na(+) by affinity. Ca(v)3.1 and Ca(v)3.2 discriminate Ca(2+), Sr(2+) and Ba(2+) based on the ion's effects on the open channel probability, whilst Ca(v)3.3 discriminates based on the ion's intrapore binding affinity. All Ca(v)3s were characterized by much smaller difference in the K(D) values for Na(+) current blockade by Ca(2+) (K(D1) approximately 6 microM) and for Ca(2+) current saturation (K(D2) approximately 2 mM) as compared to L-type channels. This enabled them to carry notable mixed Na(+)/Ca(2+) current at close to physiological Ca(2+) concentrations, which was the strongest for Ca(v)3.3, smaller for Ca(v)3.2 and the smallest for Ca(v)3.1. In addition to intrapore Ca(2+) binding site(s) Ca(v)3.2, but not Ca(v)3.1 and Ca(v)3.3, is likely to possess an extracellular Ca(2+) binding site that controls channel permeation. Our results provide novel functional tests for identifying subunits responsible for T-type Ca(2+) current in native cells. PMID- 17400182 TI - Overexpressed CacyBP/SIP leads to the suppression of growth in renal cell carcinoma. AB - Calcyclin-binding protein/Siah-1-interacting protein (CacyBP/SIP), a target protein of S100, has been identified as a component of a novel ubiquitinylation complex leading to beta-catenin degradation, which was found to be related to the malignant phenotypes of gastric cancer. However, the roles of CacyBP/SIP in renal cell carcinoma still remain unclear. In the present study, we had analyzed the expression of the CacyBP/SIP protein in human renal cancer cells and clinical tissue samples. The possible roles of CacyBP/SIP in regulating the malignant phenotype of renal cancer cells were also investigated. The results demonstrated that the expression of CacyBP/SIP was markedly down-regulated in renal cell carcinoma tissues and cell lines. Ectopic overexpression of CacyBP/SIP in A498 cells inhibited the proliferation of this cell and delayed cell cycle progression significantly, which might be related to the down-regulation of Cyclin D1 through reducing beta-catenin protein. CacyBP/SIP also suppressed colony formation in soft agar and its tumorigenicity in nude mice. Taken together, our work showed that CacyBP/SIP, as a novel down-regulated gene in renal cell carcinoma, suppressed proliferation and tumorigenesis of renal cancer cells. PMID- 17400183 TI - Both dimerization and interdomain processing are essential for caspase-4 activation. AB - A subgroup of caspase family of inflammatory caspases (-1, -4, -5, -11, and -12) play important role during cytokine maturation and inflammation but their regulation is not well understood as much as the initiator and effector caspases. Here, the biochemical mechanism of caspase-4 activation is elucidated. With citrate, a well-known kosmotrope to enhance the monomer-dimer transition, caspase 4 was activated approximately 40 times that was comparable with that of caspase-9 ( approximately 75-fold increments). The activation reaction was mainly bimolecular (n=1.67+/-0.04) for monomeric caspase-4. In addition, the interdomain cleavage was also responsible to activate caspase-4 more than 100-fold, again comparable with that of effector caspases where the proteolytic processing is considered as the sole activation mechanism. Thus, caspase-4 shows a novel activation mechanism of the synergism between dimerization and proteolysis that sharply differs from the established activation mechanism of dimerization for initiators and interdomain cleavage for effector caspases. PMID- 17400184 TI - Analyses of the structural organization of unidentified open reading frames from metagenome. AB - Although there is no sequence information, activity-based screening methods can select positive clones from a metagenomic library. However, the low frequency of positive hits that is caused by improper expression of proteins in the cloning host Escherichia coli might be improved. In order to investigate whether the metagenome can be expressed in E. coli, the structural organization of URFs from metagenome was analyzed in terms of transcription and translation factors, and compared to those of 4300 ORFs of E. coli K12. Considerable differences in amino acid composition and codon usage occurred between the metagenome URFs and E. coli ORFs, reflecting a barrier for protein expression within the host E. coli. From the analyses of the promoter and RBS regions, sequences or patterns in the corresponding region of metagenome URFs were found to be dissimilar to E. coli consensus. These results suggested that these factors are considerable to screen the clones from metagenomic library with the activity-based approach. PMID- 17400185 TI - Cell cycle-dependent mitochondrial biogenesis and dynamics in mammalian cells. AB - During cell cycle progression and division, how cells coordinate mitochondrial biogenesis, distribution, and partitions remains to be clarified. Here, we show that mitochondrial mass and mitochondrial membrane potential increased from early G(1) to G(1)/S and further gradually elevated to mitotic phase, indicating that mitochondrial biogenesis begins from early G(1) phase. In addition, mitochondrial DNA contents appeared to increase from G(1)/S to G(2) phase during which a slight but consistent increase of NRF-1 level was observed. However, other transcriptional factors regulating mitochondrial biogenesis, mtTFA and PRC, were not changed. During interphase, heterogeneous mitochondrial population with different morphology and sizes were observed but reorganized into relatively homogeneous population of mitotic cells. Moreover, microtubule and dynein complex, p150(Glued) and dynein intermediate chain, strongly associate with mitochondria during interphase but dissociated from them during mitosis. Taken together, our results suggest that mitochondrial biogenesis and dynamics are tightly regulated during cell cycle progression. PMID- 17400186 TI - Inhibitory effects of caffeine on hippocampal neurogenesis and function. AB - Caffeine is one of the most extensively consumed psychostimulants in the world. However, compared to short-term effects of caffeine, the long-term effects of caffeine consumption on learning and memory are poorly characterized. The present study found that long-term consumption of low dose caffeine (0.3 g/L) slowed hippocampus-dependent learning and impaired long-term memory. Caffeine consumption for 4 weeks also significantly reduced hippocampal neurogenesis compared to controls. From these results, we concluded that long-term consumption of caffeine could inhibit hippocampus-dependent learning and memory partially through inhibition of hippocampal neurogenesis. PMID- 17400187 TI - N-acetylchitooligosaccharide is a potent angiogenic inhibitor both in vivo and in vitro. AB - N-acetylchitooligosaccharide (N-acetyl-COs) was prepared by N-acetylation of chitooligosaccharide (COs). In vitro study using human umbilical vein endothelial cells (HUVECs) revealed that both N-acetyl-COs and COs inhibited the proliferation of HUVECs by inducing apoptosis. Treatment of HUVECs by N-acetyl COs resulted in a significant reduction of density of the migration cells and repressed tubulogenesis process. The antiangiogenic effects of the oligosaccharides were further evaluated using in vivo zebrafish angiogenesis model, and the results showed that both oligosaccharides inhibited the growth of subintestinal vessels (SIV) of zebrafish embryos in a dose-dependent manner, as observed by endogenous alkaline phosphatase (EAP) staining assay. In contrast, no cytotoxicity was found when treating the NIH3T3 and several other cancer cells with the oligosaccharides. Our results also confirmed the antiangiogenic activity of N-acetyl-COs was significantly stronger than the parent oligosaccharide, COs. PMID- 17400188 TI - Conjugated linoleic acid induces apoptosis of murine mammary tumor cells via Bcl 2 loss. AB - Conjugated linoleic acid (CLA) is a powerful anticancer agent in a number of tumor model systems; however, its precise mechanism of action remains elusive. Here, we report that t10,c12 CLA, a component of synthetic CLA supplements, induced apoptosis and G1 arrest of p53 mutant TM4t murine mammary tumor cells. Furthermore, t10,c12-CLA induced a time- and concentration-dependent cleavage of caspases-3 and -9, and release of cytochrome c from mitochondria to cytosol. Levels of Bcl-2 protein were decreased both in total cellular lysates and in mitochondria after t10,c12-CLA treatment; however, there was no significant change in Bax or Bak. Overexpression of Bcl-2 attenuated apoptosis in response to t10,c12-CLA treatment. These results demonstrate that t10,c12-CLA triggers apoptosis of p53 mutant murine mammary tumor cells through the mitochondrial pathway by targeting Bcl-2. PMID- 17400189 TI - Effects of chromosomal variations on pharmacokinetic activity of zolpidem in healthy volunteers: an array-based comparative genomic hybridization study. AB - Zolpidem has been known as a very safe and effective hypnotic drug used to treat a variety of patients with insomnia. Even though the same dose of the medicine is administered to each patient, the blood level of zolpidem and the time required to obtain peak concentration are not consistent among different people. We evaluated the relationship between the peak concentrations of zolpidem and chromosomal imbalances using a high-resolution genome-wide array-based comparative genomic hybridization (CGH) in 16 healthy volunteers in order to detect the genetic factors underlying the variations. The present study showed that chromosomal losses were detected in the 4q35.2, 9p13.1 and 9p12 regions, and those gains were indicated in the 2p14, 11q13.4 and 15q11.2 regions. The abnormal regions were confirmed by fluorescence in situ hybridization (FISH) and real-time PCR. It is suggested that array-CGH analysis may be used as a measure for pharmacogenomic applications in the patients with insomnia and for further exploration of candidate genomic regions implicated in sleep disturbances. PMID- 17400190 TI - Real-time imaging nuclear translocation of Akt1 in HCC cells. AB - Akt is one of the critical mediators in cellular signaling, and overactivation of Akt related pathway frequently occurs in hepatocellular carcinoma (HCC). In this study, we presented that Akt was upregulated in HCC cell lines, and its active phosphorylated form was mainly located in the nucleus. Employing the laser confocal techniques for imaging intracellular protein dynamics, we monitored the transnuclear movement of GFP-tagged wild-type Akt1 (Akt1-WT-GFP) and its inactive mutant (Akt1-T308A/S473A-GFP) in live SMMC-7721 HCC cells, and both of fusion proteins were found to distribute over the cytoplasm and nucleus. Moreover, it was found that platelet derived growth factor (PDGF) was able to accelerate the nuclear translocation of wild-type Akt1 in HCC cells but failed to speed up the motion of the mutant. It was demonstrated that activation of phosphatidylinositol 3-kinase (PI3K) and Akt1 facilitated the nuclear translocation of Akt1, but the phosphorylation at threonine 308 and serine 473 was not prerequisite. PMID- 17400191 TI - Action of neltenexine on anion secretion in human airway epithelia. AB - Neltenexine has been applied to human lung diseases such as chronic obstructive pulmonary disease (COPD) as a mucolytic agent. However, we have no information on the neltenexine action in bronchial epithelial cells. We studied the neltenexine action on the ion transport in human submucosal serous Calu-3 cells. Under a hyper-secreting condition caused by terbutaline (a beta2-adrenergic agonist), neltenexine diminished anion secretion by inhibiting the Cl- and HCO3- uptake via Na+/K+/2Cl- cotransporter and Na+/HCO3- cotransporter without blockade of the cystic fibrosis transmembrane conductance regulator (CFTR) channel, and also diminished anion secretion via stimulation of Cl-/HCO3- exchanger, which facilitates the extrusion of more CFTR-permeant anion, Cl-, with the uptake of less CFTR-permeant anion, HCO3-. Thus, neltenexine reduced the hyper-secretion to keep an appropriate fluid level in the airway, providing a possibility that neltenexine can be an effective drug in airway obstructive diseases by decreasing the airway resistance under a hyper-secreting condition. PMID- 17400192 TI - HIV-1 gp41 ectodomain enhances Cryptococcus neoformans binding to HBMEC. AB - Cryptococcus neoformans infection has significantly increased recently, particularly in AIDS patients and immunocompromised individuals. C. neoformans has a predilection to the brain, resulting in devastating meningoencephalitis. We have previously shown the invasion of C. neoformans into the human brain microvascular endothelial cells (HBMEC), which constitute the blood-brain barrier. Here, we demonstrated that C. neoformans invasion of HBMEC was enhanced by HIV-1 gp41 protein. Peptide mapping defined its functional domain around the disulfide-bond linkage of gp41 molecule (a.a. 579-611). Recombinant protein gp41 I90 (a.a. 550-639) can also enhance the binding activity. The enhancement of C. neoformans binding to HBMEC is a strain-independent manner, suggesting that gp41 ectodomain peptide exerts its function directly on HBMEC. Importantly, the enhancement could be observed in mouse animal model. Our results suggest that HIV 1 gp41 ectodomain and C. neoformans may follow a similar invasion mechanism, possibly actin reorganization and/or membrane activation, during pathogen infections on HBMEC. PMID- 17400193 TI - The first extracellular domain of claudin-7 affects paracellular Cl- permeability. AB - Tight junctions (TJ) constitute paracellular diffusion channels regulating the passage of ions and solutes across epithelia. We recently demonstrated that overexpression of the TJ membrane protein claudin-7 in LLC-PK1 cells decreases paracellular permeability to Cl(-) and increases paracellular permeability to Na(+). To investigate the effect of charged amino acid residues in extracellular domains (ED) of claudin-7 on paracellular charge selectivity, we created claudin 7 mutants by replacing negatively charged amino acids on ED with positively charged amino acids. Immunofluorescence light microscopy showed that these mutant proteins were correctly targeted to the cell junction. Ultrastructure examination of TJ morphology did not reveal any difference between cells expressing wildtype (WT) and mutant claudin-7. However, electrophysiological studies showed increased Cl(-) permeability in cells expressing first extracellular domain (ED1) mutants, but not second extracellular domain (ED2) mutants, compared to that of WT claudin 7. Our results demonstrate that negatively charged amino acids in ED1 of claudin 7 are involved in modulating paracellular Cl(-) permeability. PMID- 17400194 TI - DNA interactions of new cytotoxic tetrafunctional dinuclear platinum complex trans,trans-[{PtCl2(NH3)}2(piperazine)]. AB - A new tetrafunctional dinuclear platinum complex trans,trans [{PtCl2(NH3)}2(piperazine)] with sterically rigid linking group was designed, synthesized and characterized. In this novel molecule, the DNA-binding features of two classes of the platinum compounds with proven antitumor activity are combined, namely trans oriented bifunctional mononuclear platinum complexes with a heterocyclic ligand and polynuclear platinum complexes. DNA-binding mode of this new complex was analyzed by various methods of molecular biology and biophysics. The complex coordinates DNA in a unique way and interstrand and intrastrand cross-links are the predominant lesions formed in DNA in cell-free media and in absence of proteins. An intriguing aspect of trans,trans [{PtCl2(NH3)}2(piperazine)] is that, using a semi-rigid linker, interstrand cross linking is diminished relative to other dinuclear platinum complexes with flexible linking groups and lesions that span several base pairs, such as tri- and tetrafunctional adducts, become unlikely. In addition, in contrast to the inability of trans,trans-[{PtCl2(NH3)}2(piperazine)] to cross-link two DNA duplexes, the results of the present work convincingly demonstrate that this dinuclear platinum complex forms specific DNA lesions which can efficiently cross link proteins to DNA. The results substantiate the view that trans,trans [{PtCl2(NH3)}2(piperazine)] or its analogues could be used as a tool for studies of DNA properties and their interactions or as a potential antitumor agent. The latter view is also corroborated by the observation that trans,trans [{PtCl2(NH3)}2(piperazine)] is a more effective cytotoxic agent than cisplatin against human tumor ovarian cell lines. PMID- 17400195 TI - An event related functional magnetic resonance imaging study of facial emotion processing in Asperger syndrome. AB - BACKGROUND: People with Asperger syndrome (AS) have life-long deficits in social behavior. The biological basis of this is unknown, but most likely includes impaired processing of facial emotion. Human social communication involves processing different facial emotions, and at different intensities. However nobody has examined brain function in people with AS when implicitly (unconsciously) processing four primary emotions at varying emotional intensities. METHODS: We used event-related functional magnetic resonance imaging (MRI) to examine neural responses when people with AS and controls implicitly processed neutral expressions, and mild (25%) and intense (100%) expressions of fear, disgust, happiness, and sadness. We included 18 right-handed adults; 9 with AS and 9 healthy controls who did not differ significantly in IQ. RESULTS: Both groups significantly activated 'face perception' areas when viewing neutral faces, including fusiform and extrastriate cortices. Further, both groups had significantly increased activation of fusiform and other extrastriate regions to increasing intensities of fear and happiness. However, people with AS generally showed fusiform and extrastriate hyporesponsiveness compared to controls across emotion types and intensities. CONCLUSIONS: Fusiform and extrastriate cortices are activated by facial expressions of four primary emotions in people with AS, but generally to a lesser degree than controls. This may partly explain the social impairments of people with AS. PMID- 17400196 TI - Pedophilia is linked to reduced activation in hypothalamus and lateral prefrontal cortex during visual erotic stimulation. AB - BACKGROUND: Although pedophilia is of high public concern, little is known about underlying neural mechanisms. Although pedophilic patients are sexually attracted to prepubescent children, they show no sexual interest toward adults. This study aimed to investigate the neural correlates of deficits of sexual and emotional arousal in pedophiles. METHODS: Thirteen pedophilic patients and 14 healthy control subjects were tested for differential neural activity during visual stimulation with emotional and erotic pictures with functional magnetic resonance imaging. RESULTS: Regions showing differential activations during the erotic condition comprised the hypothalamus, the periaqueductal gray, and dorsolateral prefrontal cortex, the latter correlating with a clinical measure. Alterations of emotional processing concerned the amygdala-hippocampus and dorsomedial prefrontal cortex. CONCLUSIONS: Hypothesized regions relevant for processing of erotic stimuli in healthy individuals showed reduced activations during visual erotic stimulation in pedophilic patients. This suggests an impaired recruitment of key structures that might contribute to an altered sexual interest of these patients toward adults. PMID- 17400197 TI - Neural mechanisms of language comprehension: challenges to syntax. AB - In 1980, the N400 event-related potential was described in association with semantic anomalies within sentences. When, in 1992, a second waveform, the P600, was reported in association with syntactic anomalies and ambiguities, the story appeared to be complete: the brain respected a distinction between semantic and syntactic representation and processes. Subsequent studies showed that the P600 to syntactic anomalies and ambiguities was modulated by lexical and discourse factors. Most surprisingly, more than a decade after the P600 was first described, a series of studies reported that semantic verb-argument violations, in the absence of any violations or ambiguities of syntax can evoke robust P600 effects and no N400 effects. These observations have raised fundamental questions about the relationship between semantic and syntactic processing in the brain. This paper provides a comprehensive review of the recent studies that have demonstrated P600s to semantic violations in light of several proposed triggers: semantic-thematic attraction, semantic associative relationships, animacy and semantic-thematic violations, plausibility, task, and context. I then discuss these findings in relation to a unifying theory that attempts to bring some of these factors together and to link the P600 produced by semantic verb-argument violations with the P600 evoked by unambiguous syntactic violations and syntactic ambiguities. I suggest that normal language comprehension proceeds along at least two competing neural processing streams: a semantic memory-based mechanism, and a combinatorial mechanism (or mechanisms) that assigns structure to a sentence primarily on the basis of morphosyntactic rules, but also on the basis of certain semantic-thematic constraints. I suggest that conflicts between the different representations that are output by these distinct but interactive streams lead to a continued combinatorial analysis that is reflected by the P600 effect. I discuss some of the implications of this non-syntactocentric, dynamic model of language processing for understanding individual differences, language processing disorders and the neuroanatomical circuitry engaged during language comprehension. Finally, I suggest that that these two processing streams may generalize beyond the language system to real-world visual event comprehension. PMID- 17400198 TI - S100B-mediated protection against the pro-apoptotic effects of ethanol on fetal rhombencephalic neurons. AB - Previously, this laboratory demonstrated that ethanol treatment significantly reduces the number of developing serotonin (5-HT) and other fetal rhombencephalic neurons in rats by augmenting apoptosis. Using a 5-HT(1A) agonist we were able to attenuate the ethanol-associated reduction and apoptosis of 5-HT and rhombencephalic neurons. The downstream pro-survival effects of 5-HT(1A) stimulation were associated with the activation of phosphatidylinositol 3'kinase (PI-3K) and its subsequent up-regulation of specific NF-kappaB-dependent pro survival genes. Using an in vitro model, we investigated the hypothesis that S100B, a protein which is released from astrocytes following 5-HT(1A) agonist stimulation, can reduce apoptosis in ethanol-treated rat fetal rhombencephalic neurons. We also evaluated whether the anti-apoptotic effects of S100B on fetal rhombencephalic neurons were linked to the activation of the PI-3K-->pAkt pro survival pathway and the expression of two NF-kappaB-dependent pro-survival genes: XIAP and Bcl-2. Moreover, we determined whether S100B's pro-survival effects were associated with mitogen activated protein kinase kinase (MAPKK)- >p42/p44 MAPK. The results of these investigations demonstrated that S100B treatment prevented ethanol-associated apoptosis of fetal rhombencephalic neurons. In addition, it appears that these neuroprotective effects are linked to activation of the PI-3K pathways, because the PI-3K inhibitor LY294002 blocks the neuroprotective effects of S100B. Moreover, S100B increases the formation of pAkt and the up-regulation of two downstream NF-kappaB-dependent pro-survival genes: XIAP and Bcl-2. Although the MAPKK inhibitor PD98059 reduced the number of surviving neurons in S100B-treated cultures, S100B did not activate MAPKK. PMID- 17400199 TI - Vitamin A deficiency induces a decrease in EEG delta power during sleep in mice. AB - Recent report (Maret, S., Franken, P., Dauvilliers, Y., Ghyselinck, N.B., Chambon, P., Tafti, M., 2005. Retinoic acid signaling affects cortical synchrony during sleep. Science 310, 111-113.) has suggested that vitamin A (retinol and its derivatives) is genetically involved in the electroencephalogram (EEG) delta oscillation during sleep. However, this finding has not yet been confirmed by other studies. In this study, we attempted to record the sleep EEG and behavior, and to quantify striatal monoamines in mice fed a vitamin A-deficient (VAD) diet for 4 weeks, in order to clarify the linkage between the delta oscillation and vitamin A. VAD mice demonstrated a significant decrease in the delta power of the EEG. However, 6-h sleep deprivation caused the recovery of the delta power in VAD mice to a level similar to that of the control. VAD also caused the decrease of spontaneous activity throughout 24-h period. Furthermore, dihydroxyphenylacetic acid, a metabolite of dopamine, was decreased significantly in the striatal tissue of VAD mice. Our present results suggest that the deficiency of vitamin A causes the attenuation of delta power in NREM sleep and spontaneous activity. These attenuations may be related to the alteration of striatal dopaminergic function. PMID- 17400200 TI - Olfactory abnormalities in Huntington's disease: decreased plasticity in the primary olfactory cortex of R6/1 transgenic mice and reduced olfactory discrimination in patients. AB - Reduced neuronal plasticity in the striatum, hippocampus, and neocortex is a common feature of transgenic mouse models of Huntington's disease (HD). Doublecortin (DCX) and polysialylated neural cell adhesion molecule (PSA-NCAM) are associated with structural plasticity in the adult mammalian brain, are markers of newly formed neurons in the dentate gyrus of the adult hippocampus, and are highly expressed in primary olfactory (piriform) cortex. Animal studies have demonstrated that a reduction in plasticity in the piriform cortex is associated with a selective impairment in odour discrimination. Therefore, the number of DCX and PSA-NCAM immunoreactive cells in the piriform cortex were quantified as measures of plasticity in early stage (fifteen week old) R6/1 transgenic HD mice. The transgenic mice had a large reduction in the number of DCX and PSA-NCAM immunoreactive cells in the piriform cortex, similar to that previously reported in the R6/2 mice. We also tested whether odour discrimination, as well as identification and detection, were impaired in HD patients and found that patients (at a similar disease stage as the mice) had an impairment in odour discrimination and identification, but not odour detection. These results suggest that olfactory impairments observed in HD patients may be the result of reduced plasticity in the primary olfactory cortex. PMID- 17400201 TI - Intermittent activation of bradykinin B2 receptors and mitochondrial KATP channels trigger cardiac postconditioning through redox signaling. AB - OBJECTIVE: Postconditioning (PostC) maneuvers allow post-ischemic accumulation of autacoids, which trigger protection. We tested if PostC-triggering includes bradykinin (BK) B2 receptor activation and its downstream pathway. METHODS AND RESULTS: Isolated rat hearts underwent 30 min ischemia and 120 min reperfusion. Infarct size was evaluated using nitro-blue tetrazolium staining. In Control hearts infarct size was 61+/-5% of risk area. PostC (5 cycles of 10 s reperfusion/ischemia) reduced infarct size to 22+/-4% (p<0.01). PostC protection was abolished by B2 BK receptor-antagonists (HOE140 or WIN64338), nitric oxide synthase-inhibitor (L-nitro-arginine-methylester), protein kinase G (PKG)-blocker (8-bromoguanosine-3',5'-cyclic-monophosphorothioate), and mitochondrial K(ATP) (mK(ATP))-blocker (5-hydroxydecanoate) each given for 3 min only. Since 3 min of BK-infusion (100 nM) did not reproduce PostC protection, protocols with Intermittent-BK infusion were used to mimic PostC: a) 5 cycles of 10 s oxygenated no-BK/oxygenated+BK buffer; b) 5 cycles of 10 s oxygenated-no-BK/hypoxic+BK buffer. Both protocols with Intermittent-BK attenuated infarct size (36+/-5% and 38+/-4%, respectively; p<0.05 vs Control and NS vs PostC for both; NS vs each other). Intermittent-BK protection was abolished by the same antagonists used to prevent PostC protection. Intermittence of re-oxygenation only (5 cycles of 10 s oxygenated/hypoxic buffer) did not reproduce PostC. Yet, cardioprotection was triggered by intermittent mK(ATP) activation with diazoxide, but not by intermittent reactive oxygen species (ROS) generation with purine/xanthine oxidase. ROS scavengers (N-acetyl-L-cysteine or 2-mercaptopropionylglycine), given for 3 min only, abolished PostC-, Intermittent BK-and diazoxide-induced protection. CONCLUSIONS: Intermittent targeting of specific cellular sites (i.e. BK B2 receptors and mK(ATP) channels) during early reperfusion triggers PostC protection via ROS signaling. Since neither intermittent oxygenation nor exogenous ROS generators can trigger protection, it is likely that intermittent autacoid accumulation and ROS compartmentalization may play a pivotal role in PostC-triggering. PMID- 17400202 TI - New biochemical markers: from bench to bedside. AB - BACKGROUND: Evaluation of patients presenting to hospital with chest pain or other signs or symptoms suggesting acute coronary syndrome (ACS) is problematic, time-consuming and sometimes expensive, even if new biochemical markers, such as troponins, have improved the ability to detect cardiac injury. However, patients with normal troponin values are not necessarily risk-free for major cardiac events. METHODS: Recent investigations indicate that the overall patient risk may be assessed earlier than before, thanks to new knowledge acquired concerning the pathobiology of atherosclerosis and molecular events involved in the progression of disease, thus allowing the development of new biochemical markers. Some selected markers are released during the different phases of development of cardiovascular disease and may be useful for the diagnosis of patients with cardiovascular disease. In particular, the identification of emerging markers that provide relevant information on the inflammatory process, and the development of biomarkers whose circulating concentrations suggest the status of plaque instability and rupture, seems to be of particular value in prognosis and risk stratification. The overall expectations for a cardiovascular biochemical marker are not only its biological plausibility but also the availability at a reasonable cost of rapid, high quality assays, and their correct interpretation by clinicians using optimal cut-offs. CONCLUSION: The crossing from bench to bedside for each new marker discovered, must be associated with concurrent advances in the characterization of analytical features and the development of routine assay, in the assessment of analytical performance and in interpretative reporting of test results as well as in the training of physicians to use the array of biomarkers available appropriately and to interpret them correctly. This approach calls for the coordinated support of clinicians, technology experts, statisticians and the industry so that new biochemical developments can be of optimal value. PMID- 17400203 TI - Neovascularization and cardiomyocytes regeneration in acute myocardial infarction after bone marrow stromal cell transplantation: comparison of infarct-relative and noninfarct-relative arterial approaches in swine. AB - BACKGROUND: Adult bone marrow stromal cells could differentiate into myogenic endothelial progenitor cells and has been investigated for the potential value in regeneration. Recently, it has been reported that bone marrow cells (BMCs) are able to repair the infracted myocardium by intracoronary transplantation via infarct-related artery in humans. Unfortunately, we cannot open the infarcted artery by traditional reperfusion therapies in some patients. We investigate the hypothesis that BMCs transplantation might get the same effect via noninfarct relative artery. This alternative approach may have potential application in clinical practice. METHODS: A swine myocardial infarction model was established by distal left anterior descending artery ligation. Bone marrow stromal cells isolated, culture-expanded and labeled with bromodeoxyuridine (BrdU) were used as donor cells. Four weeks after coronary artery ligation, either a graft of 5x10(6) donor cells (n=12) or culture medium (n=6) was infused into infarcted area via infarct-relative artery (left coronary artery, n=6) and noninfarct-relative artery (right coronary artery, n=6). Heart function was evaluated by gate cardiac perfusion imaging before the transplantation and 4 weeks after transplantation. The donor cell localization and differentiation were identified by immunohistochemical staining for BrdU and beta-myosin heavy chain (beta-MHC) and angiogenesis was assessed by immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA) and Factor VIII. RESULTS: Gate cardiac perfusion imaging demonstrated that the cardiac function was significantly improved after the stromal cell transplantation via both infarct-relative and noninfarct-relative coronary arteries compared with control group (45.03+/-2.71 and 47.78+/-2.64 vs 30.36+/-2.76, P<0.05). Four weeks after transplantation, BrdU and beta-MHC positive cells were detected within the infarct area. Vessel densities in infarct area and infarct border area were increased significantly in both transplantation groups compared to the control group (98.68+/-5.32 and 87.49+/-6.04 vs 48.46+/ 4.88, P<0.05). CONCLUSIONS: Transplantation of bone marrow stromal cell via both infarct-relative and noninfarct-relative coronary arteries improved heart function in the myocardial infarction animals by stimulating cardiomyocyte regeneration and angiogenesis. PMID- 17400204 TI - Cryopreservation of common carp (Cyprinus carpio) sperm in 1.2 and 5 ml straws and occurrence of haploids among larvae produced with cryopreserved sperm. AB - Experiments were carried out on the cryopreservation of common carp (Cyprinus carpio) sperm in order to test the suitability of using 1.2 and 5 ml straws and to investigate the ploidy of malformed larvae found among the hatched progeny. In the first set of experiments, the effect of freezing time was investigated on the hatch rate of embryos. The highest hatch rate for 1.2 ml straws was 69+/-16% at the freezing time of 4 min, and 39+/-27% for 5 ml straws at 5 min. In the second set, the effect different egg volumes fertilized with one straw of sperm on the hatch rate and the rate of malformed larvae was investigated. The highest hatch rate with 1.2 ml straws (86+/-12%) was observed when 10 g of eggs were fertilized with one straw, whereas with 5 ml straws the hatch rate was highest (65+/-18%) when 40 g of eggs were fertilized. The highest rate of malformed larvae (15+/-9%) was found in the control, whereas the highest rate of malformed larvae among the groups fertilized with cryopreserved sperm (13+/-7%) was found in the 1x dose group fertilized with 5 ml straw. The chromosome numbers of malformed larvae were investigated and haploids were found among those hatched from eggs fertilized with cryopreserved sperm whereas only diploids were found in the controls. PMID- 17400205 TI - Role of the Onecut transcription factors in pancreas morphogenesis and in pancreatic and enteric endocrine differentiation. AB - The Onecut (OC) transcription factor HNF-6 (OC-1) is required during embryogenesis for pancreatic specification, morphogenesis and endocrine differentiation. In mammals, HNF-6 has two paralogs, OC-2 and OC-3, which share DNA-binding and transcriptional activation properties and have expression patterns that overlap with that of HNF-6. This suggested that OC-2 and OC-3 play redundant roles with HNF-6 in pancreas development. Here, we have addressed this hypothesis by analyzing the phenotype of mice knockout for the Onecut factors. We found that neither OC-2 nor OC-3 is required for pancreas specification. However, OC-2 plays partially redundant roles with HNF-6 in pancreas morphogenesis and in the differentiation of endocrine precursors. As similar molecular events drive endocrine differentiation in the pancreas and gastrointestinal tract, we also investigated if Onecut factors are involved in enteroendocrine differentiation. OC-2 and OC-3 were found to delineate specific antero-posterior regions of the gut around embryonic day 12.5. Later on, OC2 was expressed in several gut cell types, whereas OC-3 behaved as a specific marker of the enteroendocrine lineage. However, OC-2 and OC-3, alone or in combination, were dispensable for gut development and enteroendocrine differentiation. In conclusion, our data reveal partially redundant roles for HNF-6 and OC-2 in developing pancreas and identify new markers for antero-posterior patterning of the gut and for enteroendocrine differentiation. PMID- 17400206 TI - The Shh-independent activator function of the full-length Gli3 protein and its role in vertebrate limb digit patterning. AB - Anterior-posterior (A/P) limb patterning in vertebrates is determined by the counteraction between the Sonic Hedgehog (Shh) and the Gli3 transcription factor. Shh exerts its effect on Gli3 by regulating the full-length Gli3 protein processing to generate a Gli3 repressor gradient along the A/P axis of the limb. However, it is not clear whether the full-length Gli3 is an activator in vivo and plays any role in the limb patterning. Here we show that mouse limbs expressing only a Gli3 repressor form exhibit mild polysyndactyly and a partial loss of digit identity, while limbs expressing only a full-length Gli3 protein display severe polysyndactyly and a complete loss of digit identity. Interestingly, when the full-length Gli3 and the repressor are equally expressed in the limb, the digit patterning is overall normal except for an extra anterior digit. Furthermore, in the presence of one Gli3 wild type allele, a Gli3 mutant allele that expresses only the full-length form can rescue the Shh mutant digit phenotype to a great extent. The full-length Gli3 protein can also activate Shh target gene expression without Shh. Thus, our data indicate that the full-length Gli3 protein is an activator in vivo and that the ratio of the Gli3 activator to repressor, but neither the Gli3 repressor gradient nor the Gli3 activator/repressor ratio gradient, determines limb digit patterning. PMID- 17400207 TI - Introduction of antisense oligonucleotides to heat shock protein 47 prevents pulmonary fibrosis in lipopolysaccharide-induced pneumopathy of the rat. AB - Acute respiratory distress syndrome (ARDS) confers high morbidity, and in part due to pulmonary fibrosis. The 47-kDa heat shock protein 47 (HSP 47) is a collagen-specific molecular chaperone that has been shown to play a major role in the processing and secretion of procollagen. We examined the effect of antisense oligonucleotides against HSP 47 in Wistar rats with lipopolysaccharide (LPS) induced pulmonary fibrosis. These rats expressed heat shock protein (HSP) 47 and collagen in response to LPS. The distribution of HSP 47 was similar to that of collagen, and all control rats displayed pulmonary fibrosis after intratracheal administration of 20 mg/kg LPS alone. Antisense oligonucleotides (100 nmol/kg dissolved in saline) were administered with the LPS among experimental subjects. Subsequent immunoblot analysis confirmed the inhibition of HSP 47 by the administration of antisense oligonucleotides. The oligonucleotides significantly improved pulmonary fibrosis among those rats administered LPS, but the oligonucletides themselves did not produce any significant changes in the behavior or histology of the lungs among control rats. These findings suggest that HSP 47 antisense oligonucleotides improve lung fibrosis among rats with LPS induced pneumopathy. PMID- 17400208 TI - Effects of co-administration of antidepressants and monoamine oxidase inhibitors on 5-HT-related behavior in rats. AB - 5-hydroxytryptamine (5-HT) syndrome is a dangerous condition of 5-HT excess that can occur during co-administration of a monoamine oxidase (MAO) inhibitor and an antidepressant. We investigated the effects of acute administration of MAO inhibitors and subchronic administration of tricyclic and heterocyclic antidepressants, and a serotonin-noradrenaline reuptake inhibitor (SNRI) on 5-HT syndrome in rats treated with 5-hydroxytryptophan (5-HTP). The irreversible and non-selective MAO inhibitor pargyline, and the reversible and selective MAO-A inhibitor clorgyline, produced increases in 5-HT syndrome in the 5-HTP-treated rats, while subchronic co-administration of imipramine partly intensified and partly attenuated the syndrome, whereas milnacipran only attenuated the syndrome. Co-administration of mianserin partly intensified and partly attenuated the syndrome but the attenuating effect was dominant. Administration of the irreversible and selective MAO-B inhibitor selegiline did not produce any increase in 5-HT syndrome in the 5-HTP-treated rats, compared with the saline control. These data suggest that non-selective MAO and selective MAO-A inhibitors can induce 5-HT syndrome in humans when co-administered with antidepressants. Furthermore, the risk of 5-HT syndrome may be lower with the selective MAO-B inhibitor selegiline. PMID- 17400209 TI - Sympathetic nervous system mediates surgical trauma stress-induced splenocyte apoptosis in rats. AB - Surgical trauma stress has been reported to induce immunosuppression. The mechanisms involved are still unclear. The present study was designed to assess the role of the sympathetic nervous system in regulating splenocyte apoptosis induced by surgical trauma stress. Our results showed that the rats that underwent surgical trauma stress exhibited a significant reduction in splenic cellularity, the loss of splenocytes was likely mediated by apoptosis, for a substantial increase in apoptosis was observed by using DNA gel electrophoresis and TUNEL assay. At the same time, an increase in Fas(CD95/Apo-1) protein expression in splenocytes was also observed. These effects were significantly abolished by either chemical sympathectomy or beta-adrenergic receptor antagonist propranolol. The data clearly revealed that the sympathetic nervous system especially beta-adrenergic receptors was involved in surgical trauma-induced immune alterations via a mechanism of apoptotic cell death. PMID- 17400210 TI - Inhibition of ErbB2/neuregulin signaling augments paclitaxel-induced cardiotoxicity in adult ventricular myocytes. AB - Paclitaxel (Taxol) has been successfully combined with the monoclonal antibody trastuzumab (Herceptin) in the treatment of ErbB2 overexpressing cancers. However, this combination therapy showed an unexpected synergistic increase in cardiac dysfunction. We have studied the mechanisms of paclitaxel/anti-ErbB2 cardiotoxicity in adult rat ventricular myocytes (ARVM). Myofibrillar organization was assessed by immunofluorescence microscopy and cell viability was tested by the TUNEL-, LDH- and MTT-assay. Oxidative stress was measured by DCF fluorescence and myocyte contractile function by video edge-detection and fura-2 fluorescence. Treatment of ARVM with paclitaxel or antibodies to ErbB2 caused a significant increase in myofilament degradation, similarly as observed with an inhibitor of MAPK-signaling, but not apoptosis, necrosis or changes in mitochondrial activity. Paclitaxel-treatment and anti-ErbB2 reduced Erk1/2 phosphorylation. Paclitaxel increased diastolic calcium, shortened relaxation time and reduced fractional shortening in combination with anti-ErbB2. A minor increase in oxidative stress by paclitaxel or anti-ErbB2 was found. We conclude, that concomitant inhibition of ErbB2 receptors and paclitaxel treatment has an additive worsening effect on adult cardiomyocytes, mainly discernible in changes of myofibrillar structure and function, but in the absence of cell death. A potential mechanism is the modulation of the MAPK/Erk1/2 signaling by both drugs. PMID- 17400211 TI - A reference dataset for circular dichroism spectroscopy tailored for the betagamma-crystallin lens proteins. AB - Circular dichroism (CD) spectroscopy is a powerful solution technique for the study of protein secondary structure. As hierarchical euclidean clustering analyses of high quality crystallin synchrotron radiation circular dichroism (SRCD) spectral data can be separated into structural groups based solely on spectral information, the technique can potentially be improved to more accurately determine secondary structures and monitor conformational changes in crystallins. Secondary structure estimates can be determined through use of reference datasets of circular dichroism spectra from proteins with determined crystal structures. As with any empirical method, the accuracies of the analyses are dependent upon how closely the reference dataset characteristics match those of the protein to be studied. To date, crystallin proteins have not been well analysed by CD because existing reference datasets do not contain good representations of their structural characteristics. This work describes a betagamma-crystallin specific reference dataset, CRYST175, which was created solely for the study of betagamma-crystallin secondary structures. Prediction accuracy was assessed for the new dataset using several deconvolution algorithms and it was found to substantially outperform existing more general reference datasets. PMID- 17400212 TI - Brain activity affects dynamic but not static autoregulation. AB - A close correlation between neuronal activity and cerebral blood flow was documented (activation-flow coupling). The matter of an activity-related effect on autoregulation remains elusive. We modulated cortical activity by GABAergic inhibition in 10 pre-anestesized rats and compared effects on activation-flow coupling and autoregulation. Contralateral forepaw stimulation was used to assess activation-flow coupling. Somatosensory potentials as well as local flow velocity responses were recorded with surface electrodes and laser-Doppler flowmetry. Performing intermittent carotid compression we calculated from the laser-Doppler data the transient hyperemic response ratio as a parameter of dynamic cerebral autoregulation. Recovery of cerebral perfusion at the end of the compression phase served as an index of static autoregulation. Tests were performed at baseline and after four successive doses of midazolam (0.5 mg/kg), applied at 30 min intervals. Reversibility of the changes was evaluated by application of flumazenil (0.2 mg/kg). Resting flow velocity levels (162+/-52 vs. 110+/-62 U; p<0.001), evoked potentials (N2-P1-amplitude; 13+/-4 vs.6+/-3 microV; p< 0.005), and resultant flow velocity responses (28+/-12 vs. 4+/-3%; p< 0.0001) decreased after the first dose of midazolam and then remained stable. A dose-dependent decrease was found for the transient hyperemic response ratio (28+/-13 to 22+/-14 to 15+/-10 (p<0.05) to 9+/-5% (p<0.025)) but not static autoregulation. After antagonism, all changes were reversible. Dynamic but not static cerebral autoregulation depends from neuronal activity and thus metabolic demand of neurons. PMID- 17400214 TI - Metformin and fetal malformations. PMID- 17400216 TI - Trying to avoid the transmission of human immunodeficiency virus particles in sperm ejaculates. PMID- 17400217 TI - Tissue-specific transcriptional initiation of the CYP19 genes in rainbow trout, with analysis of splicing patterns and promoter sequences. AB - The rainbow trout (Oncorhynchus mykiss Walbaum) genome contains three separate CYP19 genes for distinct isoforms of cytochrome P450arom: CYP19A encoding the prevalently ovarian isoform P450aromA, and CYP19B-I and II, encoding forms I and II of the mainly cerebral variant P450aromB. RNA Ligase-Mediated 5'-Rapid Amplification of cDNA Ends analysis was used to determine the 5'-untranslated terminal regions (5'-UTRs) of the corresponding mRNAs, which are actually all expressed in the ovary, brain and gills. CYP19A is transcribed at different transcription start sites (TSSs) in each tissue, the most distal TSS being found in the brain, the intermediate one in the gills, and the proximal one in the ovary. CYP19B-I also displays tissue-specific TSSs, but transcripts undergo three distinct splicing patterns: the same pattern as previously reported for the brain and occurring also in the gills, and two novel patterns, established in the ovary and brain, which include two cryptic 3'-splice sites in intron 1, leading to the inclusion of intronic sequences of 92/94 and 66 b in the 5'-UTRs. Lastly, the CYP19B-II transcript in the ovary shows the same splicing pattern previously described for the brain. A PCR-based gene walking strategy was used to explore the promoter regions of the rainbow trout CYP19 genes, which were found to contain potential binding sites for a variety of transcription factors. PMID- 17400218 TI - Differential detection of conspecific-derived odorants by the two olfactory epithelia of the Senegalese sole (Solea senegalensis). AB - The two olfactory epithelia of members of the family Soleidae sample two distinct water sources; the upper (right) side is in contact with the open water column whilst the lower (left) side is in contact with interstitial water. To evaluate whether there are differences in the sensitivities, and therefore functional roles, of the two epithelia the olfactory activity of conspecific-derived odorants was assessed in both using the electro-olfactogram (EOG). The upper nostril was significantly more sensitive to conspecific bile fluid, intestinal fluid and mucus than the lower nostril. Crude fractionation of these samples (solid-phase extraction with C18 and C2/ENV+ cartridges) revealed that olfactory activity in each body fluid was likely due to a mixture of compounds. In each case, the upper olfactory epithelium was significantly more sensitive than the lower. Similarly, olfactory sensitivity to a range of C(24) and C(27) bile acids was greater in the upper epithelium. These results suggest that intra-specific chemical communication is mediated mainly, if not entirely, by the upper olfactory epithelium. The odorants involved, and their functional roles, remain to be established. PMID- 17400219 TI - The galaninergic enteric nervous system of pleuronectiformes (Pisces, Osteichthyes): an immunohistochemical and confocal laser scanning immunofluorescence study. AB - The galaninergic enteric nervous system of three flatfishes was studied using immunohistochemical and immunofluorescence methods. Galanin-like immunoreactivity was mainly detected within the enteric intramural neurons of the stomachs and the proximal intestines. The sole, Solea solea L. and the flounder, Platichthys flesus L. showed a similar occurrence and relative distribution of galaninergic intramural neurons. Rare nervous fibre immunoreactive to the anti-galanin serum were observed in the muscular layers of the oesophagus of the turbot, Psetta maxima L. The presence and relative abundance of galanin-like immunoreactive neurons in the remaining organs of the alimentary canal of the turbot showed a different pattern in comparison to those observed in the sole and the flounder. A galanin-like peptide was detected in nerve fibres running through the exocrine parenchyma of the pancreas of all three species. It is conceivable that the galaninergic system in these species plays a role in regulating gut muscle activity, and in controlling pancreatic secretion. Galanin and choline acetyltransferase were co-localized within the same neurons of the stomach and intestine. The result confirms the hypothesis that galanin in the gut of fish functions as a cholinergic modulator. Differently from other fish species, immunoreactive endocrine cells were not detected in the studied pleuronectiformes. PMID- 17400220 TI - Hormone cross-regulation in the tadpole brain: developmental expression profiles and effect of T3 exposure on thyroid hormone- and estrogen-responsive genes in Rana pipiens. AB - During metamorphosis, the tadpole neuroendocrine brain is a major target for the organisational effects of hormones acting via both endocrine feedback mechanisms and local hormone production. While the receptor-mediated actions of thyroid hormones in brain development have been well described, there is evidence that thyroid hormones could also be an important modulator of estrogen action during metamorphosis. To better understand hormone action and potential cross-regulation between thyroid hormone and estrogen, we examined changes in thyroid hormone receptors (TRalpha and TRbeta) and the estrogen receptor (ERalpha) in the brain of Rana pipiens throughout metamorphosis and in response to 48 h waterborne triiodothyronine (T3) exposure (0.5, 5 and 50 nM). We also measured mRNA levels of iodothyronine deiodinase (D2 and D3) and aromatase, key enzymes responsible for local synthesis and availability of thyroid hormones and estrogen, respectively. A real-time PCR strategy targeting these genes was developed using either a fluorescent dual-labelled probe- or SYBR Green I-based method. TRbeta mRNA levels were increased during development and in response to T3 exposure. Deiodinase (D2 and D3) enzymes were differentially regulated during development, but mRNA levels of both were increased with 50 nM T3 exposure. ERalpha and aromatase mRNA levels significantly increased at metamorphic climax, but whereas estrogen receptor alpha mRNA levels were increased by 50 nM T3, aromatase mRNA levels were decreased. These results (1) demonstrate that the developing amphibian brain is an important site for stage-specific thyroid hormone regulation of nuclear receptors and hormone synthesis enzymes and (2) provide the basis for further studies exploring the physiological and functional significance of the cross-regulation between thyroid status and estrogen-sensitive genes in the brain during amphibian metamorphosis. PMID- 17400221 TI - Elevated hormonal stress response and reduced reproductive output in Yellow-eyed penguins exposed to unregulated tourism. AB - The endangered, endemic Yellow-eyed penguin (Megadyptes antipodes) is one of the flagship species for New Zealand's wildlife tourism, and recently concern has been raised that tourism-related pressures may be becoming too great. We compared two neighbouring breeding areas exposed to different levels of human disturbance. Penguins at the site exposed to unregulated tourism showed significantly lower breeding success and fledging weights than those in an area visited infrequently for monitoring purposes only. High parental baseline corticosterone concentrations correlated with lower fledgling weights at both sites. Stress induced corticosterone concentrations were significantly higher at the tourist exposed site, suggesting birds have been sensitized by frequent disturbance. Consequences are likely to include reduced juvenile survival and recruitment to the tourist site, while the changed hormonal stress responses may ultimately have an effect on adult fitness and survival. For maintenance of attractive Yellow eyed penguin-viewing destinations we recommend that tourists stay out of breeding areas and disturbance at penguin landing beaches is reduced. PMID- 17400222 TI - The endocrine control of salt balance in insects. AB - An overview is given of the role of Malpighian (renal) tubules and the hindgut (ileum and rectum) in the excretory process of insects. The review focuses on the mechanism of primary urine production by Malpighian tubules and its control by neurohormones, which includes serotonin and neuropeptides resembling mammalian corticotropin-releasing factor (CRF) and calcitonin. Particular emphasis is given to in vitro studies of the effect of neurohormones on Malpighian tubule ion transport and a consideration of their likely role in the regulation of salt balance in vivo. PMID- 17400223 TI - Lysine intake affects gene expression of anabolic hormones in Atlantic salmon, Salmo salar. AB - Nutritional factors influence regulation of the growth hormone (GH) and the insulin-like growth factor (IGF) system in fish, but so far there are no published studies describing how single indispensable amino acids influence these systems. Therefore, the present study aimed to test whether lysine (Lys) intake at low (LL=2.85 g/16 gN), medium (ML=4.91 g/16 gN) and high levels (HL=9.19 g/16 gN) affected the expression of genes related to the GH-IGF system (i.e. GH receptor, GH-R, IGF-I, IGF-II, IGF binding protein 1, IGFBP-1, IGF-I receptor IGF IR) in Atlantic salmon during seawater growth phase. Salmon fed the HL diet significantly up-regulated hepatic IGF-I mRNA level by a factor of 2.2 as compared to those with medium Lys intake. In addition a significant up-regulation of 2.7-fold in muscle IGF-II mRNA was present. Low Lys intake decreased the nitrogen deposition and muscle protein accretion in fish and significantly down regulated hepatic IGFBP-1 as well as muscle GH-R and IGF-II, as compared to those fed the ML diet. mRNA of IGF-IR on the other hand was not affected by Lys intake. High Lys intake resulted in a 7-fold up-regulation of muscle IGF-II mRNA level as compared to low Lys intake, and thus might be an important local anabolic regulator in fast muscle tissue. The single indispensable amino acid Lys indeed affected signalling through the genes of IGF-I, IGFBP-1 in hepatic tissue and GH R, IGF-II in fast muscle in Atlantic salmon. Concomitantly the higher Lys intake increased nitrogen deposition to a certain level. PMID- 17400224 TI - Advancing the Challenge Hypothesis. PMID- 17400225 TI - Advances in pelvic and acetabular surgery. PMID- 17400226 TI - Preoperative planning in pelvic and acetabular surgery: the value of advanced computerised planning modules. AB - An experimental computer program for virtual operation of fractured pelvis and acetabulum based on real data of the fracture is presented. The program consists of two closely integrated tools, the 3D viewing tools and the surgeon simulation tools. Using 3D viewing tools the virtual model of a fractured pelvis is built. This procedure is performed by computer engineers. Data from CT of a real injury in DICOM format are used. With segmentation process each fracture segment becomes a separate object and is assigned a different colour. The virtual object is then transferred to the personal computer of the surgeon. Bone fragments can be moved and rotated in all three planes and reduction is performed. After reduction, fixation can be undertaken. The appropriate ostheosynthetic material can be chosen. Contouring of the plate is performed automatically to the reduced pelvis. The screws can be inserted into the plate or across the fracture. The direction and length of the screws is controlled by turning the pelvis or by making bones more transparent. The modeling of the plate in all three axes can be recorded as the exact length of the screws. There is also a simulation tool for intraoperative C-arm imaging in all directions. All the steps of the procedure are recorded and printed out. Postoperative matching of real operation and virtual procedure is also possible. We operated on 10 cases using virtual preoperative planning and found it very useful. The international study is still in progress. One case is presented demonstrating all the possibilities of the virtual planning and surgery. The presented computer program is an easily usable application which brings significant value and new opportunities in clinical practice (preoperative planning), teaching and research. PMID- 17400227 TI - Femoral head injuries: Which treatment strategy can be recommended? AB - Despite different operative and non-operative treatment regimens, the outcome after femoral head fractures has changed little over the past decades. The initial trauma itself as well as secondary changes such as posttraumatic osteoarthritis, avascular necrosis or heterotopic ossification is often responsible for severe loss of function of the afflicted hip joint. Anatomic reduction of all fracture fragments seems to be a major influencing factor in determining the outcome quality. Eight years ago we inaugurated a new surgical approach for better access and visualisation for the treatment of femoral head fractures, using the "trochanteric flip" (digastric) osteotomy. Thus inspection of the entire hip joint and accurate fragment reduction under direct visual control are possible. After good initial experiences with this operative procedure we changed our standard treatment regimen to this approach in an attempt to achieve the most accurate anatomic reduction of the femoral head in every affected patient. Between 1998 and 2006 we operated on 12 patients with femoral head fractures associated with posterior hip dislocation, using the new surgical approach. Patients were followed for 2-96 months and outcome was documented with the Merle d'Aubigne and Postel score as well as the Thompson and Epstein score. The posttraumatic formation of heterotopic bone was documented with the Brooker score. Retrospective analysis of these 12 patients showed good or excellent results in 10 patients (83.3%). The two patients with poor outcome developed an avascular necrosis of the femoral head and underwent total hip arthroplasty. Periarticular heterotopic ossification was seen in five patients. In four patients this caused a significantly reduced range of motion and was therefore considered as a posttraumatic complication. The two patients with the most severe heterotopic bone formation (Brooker III and IV) had initially sustained multiple injuries including brain injury. Comparing our results with earlier published series including our own before changing the treatment protocol, the data suggest a favorable outcome in patients with trochanteric flip (digastric) osteotomy for the treatment of femoral head fractures. PMID- 17400228 TI - Nonunions and malunions after pelvic fractures: why they occur and what can be done? AB - MATERIALS AND METHODS: Between 1987 and 2005, 55 patients were treated operatively to correct 44 malunions and 11 nonunion of the pelvic ring. These pathologies were the consequence of a nonoperative initial treatment for 38 cases, or of an inappropriate indication, such as the use of an external fixator as the definitive treatment of an unstable pelvic fracture in 15 and symphysis cerclage wiring in 2. Three patients had undergone ORIF of the lumbar spine performed by neurosurgeons, but the pelvic fractures below were ignored. On the basis of damaging mechanisms and of the main instability plane, initial lesions were classified as follows: 32 shearing lesions, 11 rotatory by antero-posterior compression, 7 by lateral compression, 5 mixed. In 23 cases the site of the posterior lesion was the sacrum, 4 of which were H fractures type; 13 were sacroiliac joint dislocations, or rotatory instability of the joint (in 2 cases the lesion was bilateral), 8 were sacroiliac dislocation fractures (crescent fractures); 7 were fractures of the iliac wing. Four patients only had pubic symphysis diastasis. Indications for surgery were pain associated with deformity or instability. Surgery was performed through a multistage procedure. Mean surgery time was 6h (range: 2-10h), with a mean blood loss of 700ml (range: 200 5000ml). Follow-up ranged from a minimum of 16 months to a maximum of 14 years (mean: 5.85 years). RESULTS: At the last follow-up, all patients but one had consolidated and were considered stable. All patients had improved walking ability. Six patients still report pain. Even if most of the deformity were corrected with a significant decrease of pre-operative symptoms achieved, deformity correction was considered satisfactory but not anatomic, in 12 patients (21%). Complications occurred in 24% of patients but most were temporary. CONCLUSIONS: The most frequent cause of pelvic malunion or nonunion was inadequate treatment. To reduce the number and the percentage of disabilities, it is necessary that specialised centres provide patients with early treatment that is adequate and definitive. PMID- 17400229 TI - Feeling our way in the dark: the psychiatric nursing care of suicidal people--a literature review. AB - Psychiatric/Mental Health nurses have a long history of being front-line carers of suicidal people, and yet the international epidemiological literature, methodological problems notwithstanding, suggests that contemporary care practices for suicidal people have much room for improvement. As a result, this paper focuses on several areas/issues of care of the suicidal person, and in so doing, critiques the extant literature, such as it is. This critique illustrates that there is a disconcerting lack of empirically induced theory to guide practice and even less empirical evidence to support-specific interventions. The paper concludes, accepting the axiomatic complexity and multi-dimensionality of suicide, and the undeniable fact that suicide is a human drama, played out in the everyday lives of people, that for Psychiatric/Mental Health nurses, caring for suicidal people must be an interpersonal endeavor; and one personified by talking and listening. PMID- 17400230 TI - Effects of implant design parameters on fluid convection, potentiating third-body debris ingress into the bearing surface during THA impingement/subluxation. AB - Aseptic loosening from polyethylene wear debris is the leading cause of failure for metal-on-polyethylene total hip implants. Third-body debris ingress to the bearing space results in femoral head roughening and acceleration of polyethylene wear. How third-body particles manage to enter the bearing space between the closely conforming articulating surfaces of the joint is not well understood. We hypothesize that one such mechanism is from convective fluid transport during subluxation of the total hip joint. To test this hypothesis, a three-dimensional (3D) computational fluid dynamics (CFD) model was developed and validated, to quantify fluid ingress into the bearing space during a leg-cross subluxation event. The results indicated that extra-articular joint fluid could be drawn nearly to the pole of the cup with even very small separations of the femoral head (<0.60mm). Debris suspended near the equator of the cup at the site of maximum fluid velocity just before the subluxation began could be transported to within 11 degrees from the cup pole. Larger head diameters resulted in increased fluid velocity at all sites around the entrance to the gap compared to smaller head sizes, with fluid velocity being greatest along the anterosuperolateral cup edge, for all head sizes. Fluid pathlines indicated that suspended debris would reach similar angular positions in the bearing space regardless of head size. Increased inset of the femoral head into the acetabular cup resulted both in higher fluid velocity and in transport of third-body debris further into the bearing space. PMID- 17400231 TI - Manipulating the edge of instability. AB - We investigate the integration of visual and tactile sensory input for dynamic manipulation. Our experimental data and computational modeling reveal that time delays are as critical to task-optimal multisensory integration as sensorimotor noise. Our focus is a dynamic manipulation task "at the edge of instability." Mathematical bifurcation theory predicts that this system will exhibit well classified low-dimensional dynamics in this regime. The task was using the thumbpad to compress a slender spring prone to buckling as far as possible, just shy of slipping. As expected from bifurcation theory, principal components analysis gives a projection of the data onto a low dimensional subspace that captures 91-97% of its variance. In this subspace, we formulate a low-order model for the brain+hand+spring dynamics based on known mechanical and neurophysiological properties of the system. By systematically occluding vision and anesthetically blocking thumbpad sensation in 12 consenting subjects, we found that vision contributed to dynamic manipulation only when thumbpad sensation was absent. The reduced ability of the model system to compress the spring with absent sensory channels closely resembled the experimental results. Moreover, we found that the model reproduced the contextual usefulness of vision only if we took account of time-delays. Our results shed light on critical features of dynamic manipulation distinct from those of static pinch, as well as the mechanism likely responsible for loss of manual dexterity and increased reliance on vision when age or neuromuscular disease increase noisiness and/or time-delays during sensorimotor integration. PMID- 17400233 TI - Principles of analytical calibration/quantification for the separation sciences. AB - Calibration is an operation whose main objective is to know the metrological status of a measurement system. Nevertheless, in analytical sciences, calibration has special connotations since it is the basis to do the quantification of the amount of one or more components (analytes) in a sample, or to obtain the value of one or more analytical parameters related with that quantity. Regarding this subject, the aim of analytical calibration is to find an empiric relationship, called measurement function, which permits subsequently to calculate the values of the amount (x-variable) of a substance in a sample, from the measured values on it of an analytical signal (y-variable). In this paper, the metrological bases of analytical calibration and quantification are established and, the different work schemes and calibration methodologies, which can be applied depending on the characteristic of the sample (analyte+matrix) to analyse, are distinguished and discussed. Likewise, the different terms and related names are clarified. A special attention has been paid to those analytical methods which use separation techniques, in relation with its effect on calibration operations and later analytical quantification. PMID- 17400232 TI - New small-scale cross-axis coil planet centrifuge. Partition efficiency and application to purification of bullfrog ribonuclease. AB - The new small-scale cross-axis coil planet centrifuge (X-axis CPC) previously designed and fabricated in our laboratory has a distinctive feature such that four separation columns of similar weight are mounted symmetrically around the rotary frame to achieve stable balancing of the centrifuge under a high revolution speed. In this column layout, neighboring columns must be rotated in the opposite direction if viewed from the center of the centrifuge to avoid twisting the interconnecting flow tubes. The effect of rotational direction of the columns on the partition efficiency was evaluated with separation of a set of test samples such as cytochrome c, myoglobin, and lysozyme using an aqueous aqueous polymer phase system composed of 12.5% (w/w) polyethylene glycol (PEG) 1000 and 12.5% (w/w) dibasic potassium phosphate under 1000 rpm of column revolution. A series of experiments was performed using a set of two diagonally located columns (connected in series) each consisting of five coiled layers of 1 mm I.D. with a total capacity of 27.0 mL. Both right- and left-handed coils were tested each under the optimized conditions for choice of mobile phase and direction of the column rotation so that the satisfactory volume of the mobile phase was retained in the column by the aid of Archimedean screw effect. The results of these studies showed that one particular combination of handedness of the coil and direction of the rotation yielded the best peak resolution for each mobile phase. In order to demonstrate the capability of the apparatus, the purification of ribonuclease (RNase) from the extract of bullfrog egg, sialic acid binding lectin (cSBL), was carried out using both organic-aqueous and aqueous-aqueous polymer phase systems. When using the 16.0% (w/w) PEG 1000-6.3% (w/w) dibasic potassium phosphate-6.3% (w/w) monobasic potassium phosphate system, cSBL was successfully separated from other proteins present in the extract while commercial RNase A was eluted at near the solvent front by the lower phase mobile. The cSBL retained its native RNase activity. The overall results demonstrated that the present new small-scale X-axis CPC is useful for the purification of bioactive compounds without loss of their native activities. PMID- 17400234 TI - P-BOSS: a new filtering method for treasure hunting in metabolomics. AB - Metabolomics is expected to boost data driven research. In biomarker discovery, powerful filtering methods to remove noise and outliers are essential for screening significant candidates from the huge volume of omic data. Here we propose a post-measurement peak filtering method (named P-BOSS) for CE electrospray ionization-time-of-flight MS (CE-TOFMS) data. Combining outlier detection method functions in parallel, we applied P-BOSS to the data using Escherichia coli knockout mutants of the tryptophan and purine biosynthesis pathways. As the result, P-BOSS showed remarkably superior performance, reducing 65% of all peaks, while leaving significant peaks. PMID- 17400235 TI - Comments on "Temporal shifting: A hidden key to the skewed peak puzzle" by S.-C. Pai and L.-Y. Chiao. AB - We show that the contradiction between the temporal and spatial peak apices disappear when the position of a skewed peak is measured with the first moment rather than the peak apex. When physical chemical parameters are to be derived from the migration times, the first moment of the peak is the crucial parameter. PMID- 17400236 TI - Separation and quantification of lanthanides in synthetic standards by capillary electrophoresis: a new experimental evidence of the systematic "odd-even" pattern observed in sensitivities and detection limits. AB - The systematic zigzag pattern of sensitivities and detection limits (LODs) of lanthanides, previously observed in mass spectrometric and chromatographic measurements, was once more investigated through the indirect photometric detection with capillary electrophoresis. Well-designed chemometric experiments were performed for the electrophoretic separation and detection of lanthanides using standards with similar concentrations. A fused silica capillary 355 mm x 75 microm was used. Complete separation for all 14 lanthanides was achieved in approximately 3 min at a capillary temperature of 15 degrees C. Indirect photometric detection at 214 nm using a voltage of +25 kV and a hydrostatic injection (100 mm for 20 s) were used. The background electrolyte used consisted of an optimum mixture of 0.004 M HIBA (as complexing agent) and 0.010 M UVCat-1 (as a UV-absorbing co-ion) with a pH 4.4. A good reproducibility in migration times (<2.7% RSD), peak areas (<3.8% RSD) and peak heights (2.7% RSD) were systematically obtained. Calibration curves based on both peak areas and peak heights (from seven replicates) were prepared using weighted least-squares regressions, which were employed for the correct estimation of individual sensitivities and LODs. For a better estimation of LODs, the lowest concentration standard was injected 30 times. A new experimental evidence of the systematic "odd-even" pattern was again observed in the lanthanide sensitivities (and therefore in LODs). The calculated sensitivities were greater for lanthanides with an odd-atomic number than for their corresponding neighboring element with an even-atomic number (i.e., (57)La-(58)Ce, (59)Pr-(60)Nd; (63)Eu-(64)Gd, etc.). Concerning the LODs, a systematic zigzag pattern was observed where the odd atomic number elements have lower LODs than the even atomic number neighbor elements (i.e., (57)La-(58)Ce; (59)Pr-(60)Nd; (63)Eu-(64)Gd, etc.). The possible origin of this "odd-even" effect is briefly discussed. Accuracy errors were less than 5% for lanthanide concentrations of three synthetic standard solutions, which were considered as "unknown" samples. PMID- 17400237 TI - Selective pressurized liquid extraction for multi-residue analysis of organochlorine pesticides in soil. AB - A selective pressurized liquid extraction (SPLE) procedure capable of performing simultaneous extraction and clean-up has been demonstrated for multi-residue analysis of organochlorine pesticides (OCPs) in soil. The final method was performed at 100 degrees C for 3 x 10 min using acetone/n-heptane (1:1, v/v). Florisil was placed inside the extraction cell downstream the sample to remove interfering compounds. Extraction of two soil samples by SPLE gave a recovery which was over 80% for beta-endosulfan, endosulfan sulfate, p,p'-DDT and p,p'-DDE compared to PLE with off-line clean-up. The same trend was observed when applying the SPLE method to a certified reference soil sample (CRM 811-050) containing 13 OCPs, where the SPLE method gave 80-90% recovery vis-a-vis the PLE method with off-line clean-up. Feasibility of the SPLE method was further demonstrated by applying it to five real soil samples collected in Ethiopia. PMID- 17400238 TI - Design of simulated moving bed processes under reduced purity requirements. AB - The design of simulated moving bed chromatography under reduced purity requirements is investigated. A parametric study indicates that all internal flow rates should be considered as free parameters in process design. This is confirmed by systematic numerical optimisations of two different design scenarios. A restrictive scenario requires complete regeneration of liquid and solid phases. In contrast, in a non-restrictive design all flow rates are optimised. The latter leads to significantly improved process performance. On the basis of the obtained observations, a simple design method for simulated moving bed processes with desired outlet purity values is derived from equilibrium theory. PMID- 17400239 TI - Ovine fetal necrobacillosis. AB - The association of Fusobacterium necrophorum with late term abortion in sheep is reported. The bacterium was not culturable, but was identified in five cases by fluorescence in-situ hybridization (FISH) with an oligonucleotide probe specifically targeting 16S rRNA in F. necrophorum. Gross lesions were found in several tissues. Histologically, placental lesions were characterized by locally diffuse infiltration of neutrophils, closely associated with abundant small Gram negative and FISH-positive rods, thrombosis and necrosis. Lesions in the fetal maternal interface were multifocal and consisted of villous necrosis and suppurative inflammation. Spread to the fetus from the placenta appeared to occur in two ways. Some fetuses had multifocal necrotizing hepatitis consistent with haematogenous spread through the umbilical vein; further dissemination to other organs occurred. Transplacental spread and infection of the fetus through the amniotic fluid was characterized by development of multifocal suppurative dermatitis and suppurative bronchopneumonia. Localization of FISH-positive bacteria in necrotic lesions was restricted to the periphery. F. necrophorum would seem to have been unrecognized previously as a cause of abortion. The value of culture-independent diagnostic methods is emphasized. PMID- 17400240 TI - Cytokine expression in paraffin wax-embedded tissues from conventional calves. AB - The cross-reactivity of antibodies against human tumour necrosis factor (TNF)alpha, interleukin (IL)-1alpha, IL-1beta and porcine IL-6, and the distribution of immunolabelled cells were evaluated on paraffin wax-embedded tissues from five healthy calves. The tissues were fixed in 10% buffered formalin or Bouin's solution and processed for structural studies and immunohistochemical studies by the avidin-biotin-peroxidase technique. Bouin's solution proved to be the more suitable fixative and Tween 20 the most effective antigen unmasking technique for increasing detectable antigenicity. Constitutive expression of TNFalpha, IL-1alpha, IL-1beta and IL-6 by different cell populations, mainly macrophage-like cells, was detected. Lymphoid organs displayed a higher presence of immunolabelled cells than did lung, liver or kidney. TNFalpha and IL-1alpha appeared as the predominant cytokines, especially in the gut-associated lymphoid tissue of the ileum and in the regional mesenteric lymph nodes. The results will facilitate investigation of the role of these cytokine-producing cells in inflammatory disease processes in calves. PMID- 17400241 TI - Reversed type of color-pattern modifications of butterfly wings: a physiological mechanism of wing-wide color-pattern determination. AB - Application of cold shock or tungstate to butterfly pupae produces a unique color pattern modification type on the adult wings, in which the color-pattern elements are dislocated toward the reduced focal elements. This modification-inducing activity has been primarily attributed to the putative cold-shock hormone (CSH) that is secreted into the hemolymph upon cold shock. Here, using a species of nymphalid butterfly Junonia almana, a new "reversed" type of the color-pattern modifications of butterfly wings was obtained by the application of heat shock or thapsigargin, a calcium-ATPase inhibitor, in which most elements were dislocated away from the enlarged focal elements. This result suggests that the endocrine secretion of CSH is sensitive to a wide range of temperature shocks, which then affects the cellular interpretation of the wing-wide positional information that is emitted from the focal locations. Ecdysteroid contributes to the wing-wide patterning primarily independently from CSH, but these two systems negatively interact with each other, probably in the intracellular signaling pathways. PMID- 17400242 TI - Interspecific antagonism and virulence in hosts exposed to two parasite species. AB - Co-infection of host organisms by multiple parasite species has evolutionary consequences for all participants in the symbiosis. In this study, we co-exposed aquatic-snails (Biomphalaria glabrata) to two of their trematode parasites, Schistosoma mansoni and Echinostoma caproni. In co-exposed snails, E. caproni prevalence was 63% compared to only 23% for S. mansoni. Co-exposed E. caproni infected snails exhibited reduced fecundity, higher mortality, and higher parasite reproduction (higher virulence) compared to hosts exposed to echinostomes alone. Conversely, co-exposed S. mansoni-infected snails released fewer parasites and produced greater numbers of eggs compared to hosts exposed to S. mansoni alone. These results suggest that co-exposure not only influences the establishment (presence or absence) of particular parasite species, but also impacts host life history, parasite reproduction, and the virulence of the interaction. PMID- 17400243 TI - Molecular evolution and structural analysis of the Ca(2+) release-activated Ca(2+) channel subunit, Orai. AB - Depletion of intracellular Ca(2+) stores evokes Ca(2+) entry across the plasma membrane by inducing Ca(2+) release-activated Ca(2+) (CRAC) currents in many cell types. Recently, Orai and STIM proteins were identified as the molecular identities of the CRAC channel subunit and the endoplasmic reticulum Ca(2+) sensor, respectively. Here, extensive database searching and phylogenetic analysis revealed several lineage-specific duplication events in the Orai protein family, which may account for the evolutionary origins of distinct functional properties among mammalian Orai proteins. Based on similarity to key structural domains and essential residues for channel functions in Orai proteins, database searching also identifies a putative primordial Orai sequence in hyperthermophilic archaeons. Furthermore, modern Orai appears to acquire new structural domains as early as Urochodata, before divergence into vertebrates. The evolutionary patterns of structural domains might be related to distinct functional properties of Drosophila and mammalian CRAC currents. Interestingly, Orai proteins display two conserved internal repeats located at transmembrane segments 1 and 3, both of which contain key amino acids essential for channel function. These findings demonstrate biochemical and physiological relevance of Orai proteins in light of different evolutionary origins and will provide novel insights into future structural and functional studies of Orai proteins. PMID- 17400244 TI - Using reaction mechanism to measure enzyme similarity. AB - The concept of reaction similarity has been well studied in terms of the overall transformation associated with a reaction, but not in terms of mechanism. We present the first method to give a quantitative measure of the similarity of reactions based upon their explicit mechanisms. Two approaches are presented to measure the similarity between individual steps of mechanisms: a fingerprint based approach that incorporates relevant information on each mechanistic step; and an approach based only on bond formation, cleavage and changes in order. The overall similarity for two reaction mechanisms is then calculated using the Needleman-Wunsch alignment algorithm. An analysis of MACiE, a database of enzyme mechanisms, using our measure of similarity identifies some examples of convergent evolution of chemical mechanisms. In many cases, mechanism similarity is not reflected by similarity according to the EC system of enzyme classification. In particular, little mechanistic information is conveyed by the class level of the EC system. PMID- 17400245 TI - Energetics-based protein profiling on a proteomic scale: identification of proteins resistant to proteolysis. AB - Native states of proteins are flexible, populating more than just the unique native conformation. The energetics and dynamics resulting from this conformational ensemble are inherently linked to protein function and regulation. Proteolytic susceptibility is one feature determined by this conformational energy landscape. As an attempt to investigate energetics of proteins on a proteomic scale, we challenged the Escherichia coli proteome with extensive proteolysis and determined which proteins, if any, have optimized their energy landscape for resistance to proteolysis. To our surprise, multiple soluble proteins survived the challenge. Maltose binding protein, a survivor from thermolysin digestion, was characterized by in vitro biophysical studies to identify the physical origin of proteolytic resistance. This experimental characterization shows that kinetic stability is responsible for the unusual resistance in maltose binding protein. The biochemical functions of the identified survivors suggest that many of these proteins may have evolved extreme proteolytic resistance because of their critical roles under stressed conditions. Our results suggest that under functional selection proteins can evolve extreme proteolysis resistance by modulating their conformational energy landscapes without the need to invent new folds, and that proteins can be profiled on a proteomic scale according to their energetic properties by using proteolysis as a structural probe. PMID- 17400246 TI - Stable complexes formed by HIV-1 reverse transcriptase at distinct positions on the primer-template controlled by binding deoxynucleoside triphosphates or foscarnet. AB - Binding of the next complementary dNTP by the binary complex containing HIV-1 reverse transcriptase (RT) and primer-template induces conformational changes that have been implicated in catalytic function of RT. We have used DNase I footprinting, gel electrophoretic mobility shift, and exonuclease protection assays to characterize the interactions between HIV-1 RT and chain-terminated primer-template in the absence and presence of various ligands. Distinguishable stable complexes were formed in the presence of foscarnet (an analog of pyrophosphate), the dNTP complementary to the first (+1) templating nucleotide or the dNTP complementary to the second (+2) templating nucleotide. The position of HIV-1 RT on the primer-template in each of these complexes is different. RT is located upstream in the foscarnet complex, relative to the +1 complex, and downstream in the +2 complex. These results suggest that HIV-1 RT can translocate along the primer-template in the absence of phosphodiester bond formation. The ability to form a specific foscarnet complex might explain the inhibitory properties of this compound. The ability to recognize the second templating nucleotide has implications for nucleotide misincorporation. PMID- 17400247 TI - Comprehensive analysis of distinctive polyketide and nonribosomal peptide structural motifs encoded in microbial genomes. AB - We developed a highly accurate method to predict polyketide (PK) and nonribosomal peptide (NRP) structures encoded in microbial genomes. PKs/NRPs are polymers of carbonyl/peptidyl chains synthesized by polyketide synthases (PKS) and nonribosomal peptide synthetases (NRPS). We analyzed domain sequences corresponding to specific substrates and physical interactions between PKSs/NRPSs in order to predict which substrates (carbonyl/peptidyl units) are selected and assembled into highly ordered chemical structures. The predicted PKs/NRPs were represented as the sequences of carbonyl/peptidyl units to extract the structural motifs efficiently. We applied our method to 4529 PKSs/NRPSs and found 619 PKs/NRPs. We also collected 1449 PKs/NRPs whose chemical structures have been determined experimentally. The structural sequences were compared using the Smith Waterman algorithm, and clustered into 271 clusters. From the compound clusters, we extracted 33 structural motifs that are significantly related with their bioactivities. We used the structural motifs to infer functions of 13 novel PKs/NRPs clusters produced by Pseudomonas spp. and Burkholderia spp. and found a putative virulence factor. The integrative analysis of genomic and chemical information given here will provide a strategy to predict the chemical structures, the biosynthetic pathways, and the biological activities of PKs/NRPs, which is useful for the rational design of novel PKs/NRPs. PMID- 17400248 TI - Modulation of MutS ATP-dependent functional activities by DNA containing a cisplatin compound lesion (base damage and mismatch). AB - DNA damage-dependent signaling by the DNA mismatch repair (MMR) system is thought to mediate cytotoxicity of the anti-tumor drug cisplatin through molecular mechanisms that could differ from those required for normal mismatch repair. The present study investigated whether ATP-dependent biochemical properties of Escherichia coli MutS protein differ when the protein interacts with a DNA oligonucleotide containing a GT mismatch versus a unique site specifically placed cisplatin compound lesion, a cisplatin 1,2-d(GpG) intrastrand cross-link with a mispaired thymine opposite the 3' platinated guanine. MutS exhibited substantial affinity for this compound lesion in hydrolytic and in non-hydrolytic conditions of ATP, contrasting with the normal nucleotide inhibition effect of mispair binding. The cisplatin compound lesion was also shown to stimulate poorly MutS ATPase activity to approach the hydrolysis rate induced by nonspecific DNA. Moreover, MutS undergoes distinct conformation changes in the presence of the compound lesion and ATP under hydrolytic conditions as shown by limited proteolysis. In the absence of MutS, the cisplatin compound lesion was shown to induce a 39 degrees rigid bending of the DNA double helix contrasting with an unbent state for DNA containing a GT mispair. Furthermore, an unbent DNA substrate containing a monofunctional adduct mimicking a cisplatin residue failed to form a persistent nucleoprotein complex with MutS in the presence of adenine nucleotide. We propose that DNA bending could play a role in MutS biochemical modulations induced by a compound lesion and that cisplatin DNA damage signaling by the MMR system could be modulated in a direct mode. PMID- 17400249 TI - Crystal structures of a quorum-quenching antibody. AB - A large number of Gram-negative bacteria employ N-acyl homoserine lactones (AHLs) as signaling molecules in quorum sensing, which is a population density-dependent mechanism to coordinate gene expression. Antibody RS2-1G9 was elicited against a lactam mimetic of the N-acyl homoserine lactone and represents the only reported monoclonal antibody that recognizes the naturally-occuring N-acyl homoserine lactone with high affinity. Due to its high cross-reactivity, RS2-1G9 showed remarkable inhibition of quorum sensing signaling in Pseudomonas aeruginosa, a common opportunistic pathogen in humans. The crystal structure of Fab RS2-1G9 in complex with a lactam analog revealed complete encapsulation of the polar lactam moiety in the antibody-combining site. This mode of recognition provides an elegant immunological solution for tight binding to an aliphatic, lipid-like ligand with a small head group lacking typical haptenic features, such as aromaticity or charge, which are often incorporated into hapten design to generate high-affinity antibodies. The ability of RS2-1G9 to discriminate between closely related AHLs is conferred by six hydrogen bonds to the ligand. Conversely, cross-reactivity of RS2-1G9 towards the lactone is likely to originate from conservation of these hydrogen bonds as well as an additional hydrogen bond to the oxygen of the lactone ring. A short, narrow tunnel exiting at the protein surface harbors a portion of the acyl chain and would not allow entry of the head group. The crystal structure of the antibody without its cognate lactam or lactone ligands revealed a considerably altered antibody combining site with a closed binding pocket. Curiously, a completely buried ethylene glycol molecule mimics the lactam ring and, thus, serves as a surrogate ligand. The detailed structural delineation of this quorum-quenching antibody will aid further development of an antibody-based therapy against bacterial pathogens by interference with quorum sensing. PMID- 17400250 TI - The impact of exposure to war stress on hospital staff: a preliminary report. PMID- 17400251 TI - Diminished rostral anterior cingulate activity in response to threat-related events in posttraumatic stress disorder. AB - BACKGROUND: Previous brain imaging studies have reported hyperactivation of the amygdala and hypoactivation of the anterior cingulate in posttraumatic stress disorder (PTSD) patients, which is believed to be an underlying neural mechanism of the PTSD symptoms. The current study specifically focuses on the abnormal activity of the rostral anterior cingulate, using a paradigm which elicits an unexpected processing conflict caused by salient emotional stimuli. METHODS: Twelve survivors (seven men and five women) of the Taegu subway fire in 2003, who later developed PTSD, agreed to participate in this study. Twelve healthy volunteers (seven men and five women) were recruited for comparison. Functional brain images of all participants were acquired using functional magnetic resonance imaging while performing a same-different judgment task, which was modified to elicit an unexpected emotional processing conflict. RESULTS: PTSD patients, compared to comparison subjects, showed a decreased rostral anterior cingulate functioning when exposed to situations which induce an unexpected emotional processing conflict. Moreover, PTSD symptom severity was negatively correlated to the level of decrease in the rostral anterior cingulate activity. CONCLUSIONS: The results of this study provide evidence that the rostral anterior cingulate functioning is impaired in PTSD patients during response-conflict situations that involve emotional stimuli. PMID- 17400252 TI - Course of cognitive functioning during the stabilization phase of schizophrenia. AB - The present study aimed at examining the longitudinal course of neuropsychological impairments in schizophrenia patients during the stabilization phase of the illness. Cognitive functioning of 151 schizophrenia patients was assessed at baseline, 9-month, and 15-month follow-up with a comprehensive battery of cognitive tests. Cognitive performance of 40 matched controls was also examined at baseline and follow-up in order to control for effects of repeated testing. We found significant improvements in memory, attention, and global cognitive functioning from baseline to 9-month follow-up. Abstraction was stable at a relatively normal level. Global cognitive functioning remained at 9-month follow-up one standard deviation below normative level. Improvements in patients' cognitive performance between the 9-month and the 15-month follow-up were fewer and less pronounced. The present study implies that schizophrenia is a static encephalopathy with trait and state dependent cognitive components particularly in the attention and memory domain. The statistically and clinically significant cognitive improvements should be ground for clinical optimism. PMID- 17400253 TI - Informed shared decision making in multiple sclerosis--inevitable or impossible? AB - Patients and health authorities increasingly claim active roles in health care decision making processes. As immune therapies in MS are partially effective MS is a prototypic condition for a shared decision making process. The treatment of acute relapses and the initiation, change or withdrawal of so called disease modifying treatments are key decisions in MS management. We developed two decision aids following the phased approach of the framework of increasing evidence for complex interventions for these key decisions. In prestudies we found that 80% of MS patients demand autonomous roles in treatment decisions which contrasts with a poor knowledge of risks. On the other hand MS patients are not disturbed by evidence-based, balanced complex information. MS patients do understand this kind of information and are able to transfer new abilities to other situations. Currently we study the effects of a 4-hour education programme on relapse management versus an information leaflet in controls in 150 MS patients. In a second trial with n=298 MS patients we study the effects of an evidence-based patient information on immunotherapy on decisional role preference and performance in the patient physician encounter. Results in early 2007 will show to which extent patient education with a focus on evidence-based patient information influences participation in the decision making process. PMID- 17400254 TI - Chronic morphine exposure increases the proportion of on-cells in the rostral ventromedial medulla in rats. AB - Chronic opiate exposure produces tolerance and hypersensitivity to mechanical and thermal stimulation that involves increased pain facilitation from the rostral ventromedial medulla (RVM). The aim of the present study was to determine the effect of sustained systemic morphine exposure on RVM neurons. Three cell types in the RVM have been described: on-cells, off-cells and neutral cells. The activity of on-cells increases in response to noxious stimulation, whereas the activity of off-cells decreases following noxious stimulation. Neutral cells remain relatively unaffected. In lightly anesthetized rats, systematic exploration throughout the RVM using single-unit extracellular recordings was used to examine both the relative proportion and the neuronal properties of the different cell classes in chronic morphine and placebo treated animals. Seven days after implanting either morphine (150 mg, s.c.) or placebo pellets a total of four electrode penetrations through the RVM were made in each animal at identical coordinates along midline. Neuronal responses related to radiant heat evoked paw withdrawals were recorded. When compared to placebo treated rats, chronic morphine increased the number of on-cells and decreased the number of neutral cells, while the number of off-cells remained unchanged. Chronic morphine exposure had no effect on the spontaneous or heat-evoked discharges in on-, off-, or neutral cells. These results indicate that chronic morphine may sensitize a subpopulation of RVM neurons to noxious stimulation, which would be expected to increase descending facilitation and promote tolerance and chronic morphine induced paradoxical pain. PMID- 17400255 TI - Arachidonic acid promotes glutamate-induced cell death associated with necrosis by 12- lipoxygenase activation in glioma cells. AB - Glutamate induced glutathione (GSH) depletion in C6 rat glioma cells, which resulted in cell death. This cell death seemed to be apoptosis through accumulation of reactive oxygen species (ROS) or hydroperoxides representing cytochrome c release from mitochondria and internucleosomal DNA fragmentation. A significant increase of 12-lipoxygenase enzyme activity was observed in the presence of arachidonic acid (AA) under GSH depletion induced by glutamate. AA promoted the glutamate-induced cell death, which reduced caspase-3 activity and diminished internucleosomal DNA fragmentation. Furthermore, AA reduced intracellular NAD, ATP and membrane potentials, which indicated dysfunction of the mitochondrial membrane. Protease inhibitors such as N-alpha-tosyl-L phenylalanine chloromethyl ketone (TPCK) and 3, 4-dichloroisocumarin (DCI) but no Ac-DEVD, a caspase inhibitor, suppressed the glutamate-induced cell death. AA reduced the inhibitory effect of TPCK and DCI on the glutamate-induced cell death. These results suggest that AA promotes cell death by inducing necrosis from caspase-3-independent apoptosis. This might occur through lipid peroxidation initiated by ROS or lipid hydroperoxides generated during GSH depletion in C6 cells. PMID- 17400256 TI - The immunomodulation of endotoxin-induced acute lung injury by hesperidin in vivo and in vitro. AB - To investigate the modulation of lung local immune responses of hesperidin (HES) on the acute lung inflammation induced by LPS in vivo. Mice were challenged with intratracheal lipopolysaccharide (100 microg) 30 min before with treatment hesperidin (200 mg/kg oral administration) or vehicle. After 4 and 24 h, bronchoalveolar lavage fluid was obtained to measure proinflammatory (TNF-alpha, IL-1 beta, IL-6), anti-inflammatory (IL-10, IL-4, IL-12) cytokines, chemokines (KC, MCP-1 and MIP-2), total cell counts, nitric oxide production, and proteins. Lung histology was performed in inflated-fixed lungs. Hesperidin downregulate the LPS-induced expression of TNF-alpha, IL-1 beta, IL-6, KC, MIP-2, MCP-1, and IL 12. It also enhanced the production of IL-4, IL-10. Total leukocyte counts; nitric oxide production, iNOS expression, and proteins were significantly decreased by hesperidin. In vitro, HES suppressed the expression of IL-8 on A549 cells and THP-1 cells, the expression of TNF-alpha, IL-1 beta, and IL-6 on THP-1 cells, the expression of ICAM-1 and VCAM-1 on A549 cells which effect cell adhesion function. The suppression of those molecules is controlled by NF-kappaB and AP-1, which are activated by I kappa B and MAPK pathways. HES inhibits those pathways, thereby suppressing the expression of IL-8, TNFalpha, IL-1 beta, IL-6, IL-12, ICAM-1 and VCAM-1. This study indicates that HES had a markedly immunomodulatory effect in a clinically relevant model of ARDS. Nevertheless, further investigations are required to determine the potential clinical usefulness of HES in the adjunctive therapy of ARDS. PMID- 17400257 TI - Sensitivity of different endpoints for in vitro measurement of genotoxicity of extractable organic matter associated with ambient airborne particles (PM10). AB - Sensitivity and correlations among three endpoints were evaluated to assess the genotoxic potential of organic complex mixtures in vitro. This study was focused on DNA adduct formation, DNA single strand break induction and tumour suppressor p53 protein up-regulation produced by extractable organic matter (EOM) absorbed on respirable particulate matter PM(10) (particulate matter<10microm) collected in three European cities (Prague, Sofia, Kosice) during winter and summer period. To compare the sensitivity of particular endpoints for in vitro measurement of complex mixture genotoxicity, the metabolically competent human hepatoma cell line Hep G2 was treated with equivalent EOM concentration of 50microg/ml. Cell exposure to EOMs resulted in significant DNA adduct formation and DNA strand break induction, however, a lack of protein p53 up-regulation over the steady state level was found. While the maximum of DNA strand breaks was determined after 2h cell exposure to EOMs, 24h treatment interval was optimal for DNA adduct determination. No substantial location- and season-related differences in EOM genotoxicity were detected using DNA strand break assessment. In agreement with these results no significant variation in DNA adduct levels were found in relation to the locality and season except for the monitoring site in Prague. The Prague EOM sample collected during summer period produced nearly three-fold lower DNA adduct level in comparison to the winter EOM sample. Comparable results were obtained when the ambient air genotoxicity, based on the concentration of carcinogenic PAHs in cubic meter of air (ng c-PAHs/m(3)), was elicited using either DNA adduct or strand break determination. In general, at least six-fold higher genotoxicity of the winter air in comparison to the summer air was estimated by each particular endpoint. Moreover, the genotoxic potential of winter air revealed by DNA adduct assessment and DNA strand break measurement increased in the same order: Kosice<50% Ca125 decrease from surgery and further chemotherapy whereas in those with no response, the corresponding survival estimate was 44.02 months (95% CI: 33.26-54.79). CONCLUSION: The lack of Ca125 response from neoadjuvant chemotherapy is not an independent prognostic factor. All patients treated with neoadjuvant chemotherapy should undergo radical debulking surgery. PMID- 17400285 TI - Overexpression of folate binding protein and mesothelin are associated with uterine serous carcinoma. AB - PURPOSE: Folate receptor alpha (FOLR1) is a membrane bound receptor involved in the transport of folate as well as other regulatory cellular processes. The purpose of this study was to examine the expression of FOLR1 in uterine cancers and to identify changes in gene expression that are associated with overexpression of FOLR1. EXPERIMENTAL DESIGN: Fifty-eight frozen uterine cancer specimens were stained for FOLR1 using immunohistochemistry and results were correlated with transcript expression noted on quantitative PCR. Total RNA from 16 cases of uterine serous carcinoma (USC) was analyzed for gene expression using the Affymetrix HG-U133A and HG-U133B GeneChip set. USCs overexpressing FOLR1 were compared to cancers with an absence of FOLR1 using binary comparison and template matching of data was used to identify genes that correlate with FOLR1 expression. Selected targets from this analysis were evaluated by quantitative PCR as well as in an independent set of USC represented in quadruplicate on a tissue microarray (TMA). RESULTS: Overexpression of FOLR1 was observed in 11/16 (69%) of USC and 0/10 normal endometrium cases using frozen tissue specimens. Binary comparison between FOLR1 positive and negative cases identified 121 genes altered by 2-fold at p<0.01 of which 45 are well correlated with FOLR1 expression pattern. Using quantitative PCR, both mesothelin (MSLN) and PTGS1 (COX1) were significantly increased in FOLR1 overexpressing tumors (p=0.014 and p=0.006 respectively). TMA confirmed that overexpression of FOLR1 and MSLN respectively occurred in 23/48 (48%) and 17/54 (32%) of pure USC. CONCLUSION: Both FOLR1 and MSLN are cell surface targets that are co-expressed at high levels in USC and are appealing targets for biologic therapy. PMID- 17400286 TI - Australian adolescents' sun protection behavior: who are we kidding? AB - PURPOSE: To examine the influence of tan preference and skin type on sun protection behaviors of Australian adolescents. METHODS: The Australian Secondary School Alcohol and Drug Questionnaires were conducted in 1993, 1996, 1999 and 2002 among randomly selected students aged 12--17. Schools were randomly selected from each education sector in each state. The self-administered questionnaire contained questions about tan preferences, skin type and usual SunSmart behavior (use of sunscreen, hats and covering clothing). RESULTS: The routine use of SunSmart behavior was low in all survey years. There was a significant decrease over time in the proportion of students who practiced SunSmart behavior, with prevalence rates lower in 2002 than in any other survey year (males: p<0.01 and females: p<0.01). As desire for a tan increased, routine practice of SunSmart behaviors decreased. Across the four survey periods, male (p<0.01) and female (p<0.01) students who preferred no tan were significantly more likely to practice SunSmart behavior than students who preferred any sort of tan. Across the four survey years, male (p<0.01) and female (p<0.01) students with skin that 'just burns' were most likely to routinely practice SunSmart behavior. CONCLUSIONS: Sun protection practices among adolescents have continued to decline significantly over time. Future educational programs require an innovative approach to modify adolescent behaviors in relation to sun exposure and sun protection. PMID- 17400287 TI - Effect of ICSI and embryo biopsy on embryo development and apoptosis according to oocyte diameter in prepubertal goats. AB - ICSI and embryo biopsy are routine methods used for assisted reproduction. However, their impact on embryo quality is still poor studied. Moreover, oocyte size is also a crucial factor for blastocyst production. In this study effect of oocyte size, ICSI and embryo biopsy was assessed in terms of incidence of apoptosis and blastocyst development. IVM-oocytes from prepubertal goats were fertilized by ICSI or IVF. Embryos obtained were divided depending on oocyte size, biopsied at day-4 post-insemination/injection and cultured for additional 4 5 days. Apoptotic cell number was assessed by TUNEL staining in day-4 embryos and blastocysts obtained. In each diameter group, ICSI did not affect embryo development, blastocyst cell number and embryo apoptotic grade in comparison to IVF. Embryo biopsy did not affect blastocyst rate and apoptotic cell number, but decreased blastocyst cell number (P=0.0018). Moreover, there was a negative relationship between blastocyst cell number and apoptotic grade (P<0.05). In conclusion, ICSI and embryo biopsy do not have negative effect on embryo quality and development. However, oocyte size has a positive relationship on blastocyst yield and quality. PMID- 17400288 TI - Tocopherol transfer protein deficiency modifies nuclear receptor transcriptional networks in lungs: modulation by cigarette smoke in vivo. AB - Dietary factors and environmental pollutants initiate signaling cascades that converge on AhR:Nrf2:NF-kappaB transcription factor (TF) networks and, in turn, affect the health of the organism through its effects on the expression of numerous genes. Reactive oxygen metabolites (ROMs) have been hypothesized to be common mediators in these pathways. alpha-Tocopherol (AT) is a potent, lipophilic, scavenger of ROMs in vitro and has been hypothesized to be a major chain-breaking anti-oxidant in lipoproteins and biological membranes in vivo. The lung offers a vital organ to test the various postulated actions of AT in vivo. Lung AT concentrations can be manipulated by several methods that include dietary and genetic techniques. In this study we have used mice with severe AT deficiency inflicted at birth by the deletion of AT transfer protein (ATTP) which is abundantly expressed in the liver and regulates systemic concentrations of AT. Mice and humans deficient in ATTP are AT deficient. Female ATTP-deficient (ATTP KO) mice and their congenic ATTP normal (WT) mice fed a diet containing 35 IU AT/kg diet were used to test our hypothesis. The mice (n=5/group) were exposed to either air or cigarette smoke (CS, total suspended particles 60 mg/m(3), 6h/day), a source of ROM, for 3 or 10 days. Post-exposure lung tissue was dissected, RNA extracted from each lung and it was pooled group-wise and processed for GeneChip analysis (Affymetrix 430A 2.0). Differential analysis of the transcriptomes ( approximately 16,000 mRNAs) identified CS sensitive genes that were modulated by lung AT-concentration. CS activated AhR driven genes such as cyp1b1 whose induction was augmented in CS-exposed, AT-deficient lungs. However, CS-induced expression of some of the Nrf2 driven genes was not potentiated in the AT deficient lungs. Largest clusters of CS-AT sensitive genes were lymphocyte and leukocyte specific genes. These gene-clusters included those encoding cytokines and immunoglobulins, which were repressed by CS and were modulated by lung AT concentrations. Our genome-wide analysis suggests reciprocal regulation of xenobiotic and immune response genes by CS and a modulatory role of lung AT concentration on the expression of these clusters of genes. These data suggest that in vivo network of AT, AT-metabolites and ATTP affects the transcription of genes driven by AhR, Nrf2 and NF-kappaB, transcription factor networks that transduce cellular metabolic signals and orchestrate adaptive responses of lungs to inhaled environmental pollutants. PMID- 17400289 TI - Processes that contribute to resilience among youth in foster care. AB - The objective of this qualitative research is to better understand the processes that contribute to resilience among adolescents in foster care. Twelve boys and girls (X=15.9 years), identified as resilient, participated in this study. The mean duration of the teenagers' placement is 7.3 years. The results point to three types of turning points: action, relation and reflection. Four processes, directly or indirectly linked to the turning point, have also been identified: increase in perceived self-efficacy, distancing oneself from the risks, new opportunities, and the multiplication of benefits. PMID- 17400290 TI - Phase transformation of plasma-sprayed hydroxyapatite coating with preferred crystalline orientation. AB - Highly oriented hydroxyapatite coatings (HACs) were obtained on titanium substrates through a radio-frequency thermal plasma spraying (TPS) method. XRD patterns showed that the HACs had crystallites with [001] preferred orientation vertical to the coating's surface. XRD results also indicated that tetracalcium phosphate crystallites in the as-sprayed HAC were oriented in the (100) direction. XRD peaks corresponding to tetracalcium phosphate, tricalcium phosphate and calcium oxide were absent after heat and hydrothermal treatment. The orientation degree of the HAC was influenced little by such post-heat treatments. Considering the crystallographic relationship between the tetracalcium phosphate in the as-sprayed HAC and the HA crystallites formed in the heat-treated HAC, these XRD results indicate that the tetracalcium phosphate in the as-prepared coatings transformed topotaxially into HA during the post-heat treatment. TEM and SEM analyses of the highly oriented HAC were conducted. The characteristic lamellar structure of TPS deposits was observed in cross-sections of the HAC. A prismatic texture was also observed in magnified SEM images. TEM observation showed that 200-800-nm-wide prismatic crystallites were formed in HA splats, and their longitudinal axis was oriented vertically to the coating's surface. SAD patterns showed that the longitudinal axis of the prismatic crystallites corresponded to the [001] zone axis of the HA crystal. PMID- 17400291 TI - Bradykinin into amygdala induces thermal hyperalgesia in rats. AB - Bradykinin is one of the most potent endogenous algesic substances and its role in pain transmission has been intensively studied in the periphery. However, the action of this peptide in central structures involved in pain transmission remains unclear. Administration of bradykinin (0.25 nmol/site) into the right amygdala of adult male Wistar rats induced thermal hyperalgesia, evaluated in the paw-flick test. Bradykinin-induced hyperalgesia was abolished by co administration with the B(2) receptor antagonist Hoe 140 (5 pmol/site), the NMDA antagonist MK-801 (5 nmol/site), the cyclooxygenase inhibitor indomethacin (10 nmol/site) and the glial metabolic inhibitor fluorocitrate (1 nmol/site). Since the intra-amygdala administration of bradykinin did not alter spontaneous locomotion in the open-field test, it is unlikely that the current described hyperalgesic effect of bradykinin is due to an unspecific action on motor activity. These findings provide evidence that bradykinin, through activation of amygdalar B(2) receptors induces hyperalgesia and that glutamatergic- and prostanoid-mediated mechanisms are involved in such effect. PMID- 17400292 TI - Bacterial infection of fly ovaries reduces egg production and induces local hemocyte activation. AB - Morbidity, the state of being diseased, is an important aspect of pathogenesis that has gone relatively unstudied in fruit flies. Our interest is in characterizing how bacterial pathogenesis affects various physiologies of the fly. We chose to examine the fly ovary because we found bacterial infection had a striking effect on fly reproduction. We observed decreased egg laying after bacterial infection that correlated with increased bacterial virulence. We also found that bacteria colonized the ovary in a previously undescribed manner; bacteria were found in the posterior of the ovary, adjacent to the lateral oviduct. This local infection in the ovary resulted in melanization and activation of the cellular immune response at the site of infection. PMID- 17400293 TI - Intrastromal corneal ring segment implantation by femtosecond laser for keratoconus correction. AB - PURPOSE: To report the outcomes after the implantation of intrastromal corneal ring segments (KERARING) aided by femtosecond laser for the correction of keratoconus. DESIGN: Prospective consecutive interventional study. PARTICIPANTS: Twenty-one eyes of 16 consecutive keratoconic patients with mean age 32+/-10.2 years. Forty-five percent were keratoconus grade I, 20% grade II, 15% grade III, and 20% grade IV. METHODS: Intrastromal corneal ring segments KERARING were implanted for keratoconus correction after corneal tunnel creation with the aid of femtosecond laser (IntraLase Corp, Irvine CA). MAIN OUTCOME MEASURES: Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), refractive outcome, complete ophthalmologic examination, videokeratography, and corneal higher order aberrations (HOA) were determined or performed before and after surgery at 1 day, 1 week, and 3 and 6 months. In addition, operative and postoperative complications were recorded. RESULTS: KERARING implantation significantly increased UCVA from 0.06 to 0.3 (P< or =0.0001), BSCVA from 0.54 to 0.71 (P< or =0.0003), and decreased the spherical equivalent by 2.28 diopters (D) and the average keratometric values (K value) by 2.24 D (P<0.001). There was no significant difference between the 3 and 6 months follow-up. Forty percent (8 eyes with a relatively low RMS of total HOA) showed a nonsignificant increase in the RMS of total HOA and 60% (12 eyes with a relatively higher RMS of total HOA> or =3.0 microm) showed significant (P< or =0.01) decrease in the RMS of total HOA due to a significant (P< or =0.003) reduction in coma and comalike aberrations. The amount of change in corneal curvature (biomechanical response) ranged from a decrease of 18% from its initial value to an increase of 2.75% and was not correlated to any preoperative parameter. Localized infectious keratitis occurred in only 1 eye (4.8%), and incision opacification occurred in 8 eyes (38%). CONCLUSION: Intrastromal corneal ring segment implantation with femtosecond laser is an effective method for correcting keratoconus with reduction in corneal HOA in eyes with coma aberration > 3.0 microm. PMID- 17400294 TI - Elevated C-reactive protein levels in patients with polypoidal choroidal vasculopathy and patients with neovascular age-related macular degeneration. AB - PURPOSE: To determine the relationship between systemic C-reactive protein (CRP) levels and polypoidal choroidal vasculopathy (PCV) and advanced neovascular age related macular degeneration (AMD) in Japanese patients. DESIGN: Case-control study. PARTICIPANTS: Ninety-seven patients with PCV, 176 with advanced neovascular AMD, and 262 control subjects without any macular abnormality were studied. METHODS: Color fundus photographs of the macular area were taken from both eyes in all subjects. Indocyanine green angiography and fluorescein angiography were performed for diagnosis. The CRP level was measured by a high sensitivity assay using a latex aggregation immunoassay, and the levels in patients with PCV and neovascular AMD were compared with that in the control group using the Kruskal-Wallis test. Associations between CRP and PCV or neovascular AMD were compared using logistic regression analysis by computing the odds ratios (ORs) and 95% confidence intervals (CIs) after the study populations were divided into quartiles. MAIN OUTCOME MEASURES: The CRP levels in patients with PCV, patients with neovascular AMD, and control subjects. Standard univariate and multivariate analyses between groups. RESULTS: Median CRP levels were significantly higher in cases with PCV (0.94 mg/l) or with advanced neovascular AMD (0.95 mg/l) than in control subjects (0.43 mg/l) (P<0.001 for Kruskal-Wallis test). After adjusting for baseline characteristics such as age, gender, smoking status, alcohol use, body mass index, history, and use of antiinflammatory drugs, the increase in risk was significant for the highest quartile of CRP for both PCV (OR, 3.53; 95% CI, 1.49-8.40) and neovascular AMD (OR, 4.08; 95% CI, 1.94-8.56), and for the third quartile of CRP for neovascular AMD (OR, 2.29; 95% CI, 1.07-4.91). The trends for an increase in risk of disease with increase in CRP were statistically significant for both PCV (P = 0.001) and neovascular AMD (P<0.001). CONCLUSIONS: The significant associations between elevated serum CRP levels and PCV or neovascular AMD in the Japanese strongly suggest that inflammatory processes are involved in the pathogenesis of PCV and neovascular AMD. PMID- 17400295 TI - Study on the isomerism in meso-amavadin and an amavadin analogue. AB - An X-ray crystallographic study of 'meso-amavadin' revealed that in the crystal the negatively charged anionic species of the title compound join into infinite hydrogen-bonded chains, counterbalanced by cationic hydronium species. Along with water of crystallization a three-dimensional hydrogen-bonded network is formed. Based on NMR- and X-ray data of amavadin and 'meso-amavadin', a model was developed that accounts for the structure of amavadin-type complexes, i.e. vanadium(IV) non-oxo complexes that contain two ligands with a tridentate N hydroxyiminodiacetate backbone. The model describes the different arrangements of the two ligands around the vanadium and it accounts for eventual symmetry in the complex. The model was used for the interpretation of NMR-data of an amavadin analogue with a benzyl group at the ligand backbone. PMID- 17400296 TI - Appropriate dosing regimens for treating juvenile rats with desipramine for neuropharmacological and behavioral studies. AB - The tricyclic antidepressants, including desipramine (DMI), are no better than placebo in treating childhood and adolescent depression, but are effective in adult depression. Animal studies comparing the effects of DMI in juveniles and adults are complicated by age-related variations in elimination rates. Thus, different dosing regiments are needed to achieve similar brain drug levels in juvenile and adult rats. We compared the half-life of DMI as well as the brain and serum concentrations of DMI and its active metabolite desmethyldesipramine in juvenile and adult rats after various drug administration paradigms. After acute i.p. administration DMI is eliminated from the brain more slowly in postnatal day (PND) 21 and 28 rats as compared to adults. After chronic i.p. administration (for 4-5 days between PND 9 and 28), lower doses of DMI are needed with juvenile rats to obtain the same brain DMI concentrations as adults. By contrast, 2 weeks of continuous drug delivery (minipump) to PND 21-35 and adult rats result in similar brain DMI concentrations. Thus, the pharmacokinetic properties of DMI varies with the age of the animal and dosing of DMI and needs to be carefully adjusted in order to have appropriate brain levels of the drug. PMID- 17400297 TI - Genomics in multiple sclerosis--current state and future directions. AB - Microarray-based gene expression profiling of large numbers of genes or even the whole genome has only recently become possible. Several studies have employed this technology in multiple sclerosis (MS) and its animal model, experimental allergic encephalomyelitis (EAE), and although results are promising, microarray based genomics research is still viewed with skepticism. It is often negatively perceived as a fishing expedition rather than a discovery-oriented effort that takes into account the immense complexity of diseases such as MS. Besides these conceptual concerns, technical reproducibility and the strategies to analyze and interpret the massive amounts of data present problems that can cause considerable variability between studies. In this review, we summarize existing data from different gene expression profiling studies that have been conducted in MS and EAE, discuss potential problems and propose future directions for the use of microarrays in MS. PMID- 17400298 TI - Genotype differentiating RT-PCR and sandwich ELISA: handy tools in epidemiological investigation of foot and mouth disease. AB - Years of molecular epidemiological surveillance has revealed co-circulation of two antigenically divergent genotypes of foot and mouth disease virus serotype A in India. Genotype differentiating RT-PCR and sandwich ELISA were developed as fast, cost-effective and user-friendly alternatives to 1D region based phylogeny for detection and differentiation of genotype VI and VII. The RT-PCR assay targeting 1D region was found to be more sensitive and authentic in distinguishing genotypes than sandwich ELISA. These assays promise to be reliable tools in the epidemiological investigation of foot and mouth disease in the country. PMID- 17400299 TI - Regulation of firing of dopaminergic neurons and control of goal-directed behaviors. AB - There are several brain regions that have been implicated in the control of motivated behavior and whose disruption leads to the pathophysiology observed in major psychiatric disorders. These systems include the ventral hippocampus, which is involved in context and focus on tasks, the amygdala, which mediates emotional behavior, and the prefrontal cortex, which modulates activity throughout the limbic system to enable behavioral flexibility. Each of these systems has overlapping projections to the nucleus accumbens, where these inputs are integrated under the modulatory influence of dopamine. Here, we provide a systems oriented approach to interpreting the function of the dopamine system, its modulation of limbic-cortical interactions and how disruptions within this system might underlie the pathophysiology of schizophrenia and drug abuse. PMID- 17400300 TI - Dopamine dysregulation of movement control in L-DOPA-induced dyskinesia. AB - The nigrostriatal dopamine (DA) system has an essential role in the selection and control of movement sequences, and its degeneration causes the characteristic motor symptoms of Parkinson's disease. Parkinsonian motor symptoms are alleviated by L-DOPA, but this treatment induces motor fluctuations and dyskinesias (abnormal involuntary movements). Clinical and experimental findings indicate that the motor complications of L-DOPA pharmacotherapy are triggered by transient and large changes in extracellular DA levels. The disruption of presynaptic DA homeostasis sets in motion a cascade of postsynaptic alterations, which prime the brain for a complicated motor response to dopaminomimetic treatment. L-DOPA induced dyskinesia provides a paradigm to study how the dysregulation of DA release and clearance results in maladaptive neuroplasticity sustaining abnormal patterns of movement. PMID- 17400301 TI - Behavioral dopamine signals. AB - Lesioning and psychopharmacological studies suggest a wide range of behavioral functions for ascending midbrain dopaminergic systems. However, electrophysiological and neurochemical studies during specific behavioral tasks demonstrate a more restricted spectrum of dopamine-mediated changes. Substantial increases in dopamine-mediated activity, as measured by electrophysiology or voltammetry, are related to rewards and reward-predicting stimuli. A somewhat slower, distinct electrophysiological response encodes the uncertainty associated with rewards. Aversive events produce different, mostly slower, electrophysiological dopamine responses that consist predominantly of depressions. Additionally, more modest dopamine concentration fluctuations, related to punishment and movement, are seen at 200-18,000 times longer time courses using voltammetry and microdialysis in vivo. Using these responses, dopamine neurotransmission provides differential and heterogeneous information to subcortical and cortical brain structures about essential outcome components for approach behavior, learning and economic decision-making. PMID- 17400302 TI - Identification of Herpes TATT-binding protein. AB - The regulation of viral gene expression is a compilation of virus and host factors influencing the transcription machinery. In Epstein-Barr Virus (EBV) a distinct regulatory element utilizing the TATT-box was described. The motif is present in promoters of lytic cycle genes and resembles a crucial host genome motif (TATA-box). Since the binding specificity of eukaryotic proteins recognizing TATA-box (TBP) was determined and no specific preference for interaction with TATT motif was found, we performed a genome-wide fold recognition search to identify viral proteins potentially recognizing the TATT box. By applying profile-profile comparisons and homology-based protein structure prediction we identified a protein of unknown function from Gammaherpesvirinae (BcRF1 of EBV) and their Betaherpesvirinae homologs (UL87 of CMV) as proteins encoding TBP fold. Although overall sequence identity is very low (circa 10%), the saddle-like fold and presence of important residues on a surface of DNA protein interface marked both proteins as distantly related to TBP and permitted the characterization of a putative molecular basis of selective recognition of TATT-motif by BcRF1. PMID- 17400303 TI - Adefovir dipivoxil treatment of lamivudine-resistant chronic hepatitis B. AB - Adefovir dipivoxil (ADV)-resistant mutations have been identified in treating hepatitis B virus (HBV) infection. This study aimed to analyze the response, the incidence of ADV resistance and the virologic characteristics of ADV therapy. A total of 29 CHB patients with confirmed lamivudine (LAM)-resistant HBV were treated with ADV for more than 52 weeks. Serum HBV DNA, HBV genotypes and sequences of HBV polymerase reverse-transcriptase domain were determined. Rates for the biochemical response, HBeAg loss, HBeAg seroconversion and virologic response (< 200 copies/mL of HBV DNA) were 82.8, 23.5, 11.8, and 48.3%, respectively, at week 52 of treatment. Lower pre-treatment mean HBV DNA level was the only significant factor associated with negative HBV DNA after ADV therapy. Six (20.7%) patients had clearance of LAM-resistant YMDD variants with replacement by the wild type HBV at week 52. The rtN236T, rtA181V/T and rtI233V were not identified before ADV therapy and the genotypic mutation of rtN236T was detected in one (3.4%) patient. In conclusion, the 52-week ADV treatment for patients with LAM-resistant HBV variants significantly achieved normalization of ALT levels, reduced serum HBV DNA levels and induced HBeAg loss and seroconversion. The emergence of ADV-resistant mutations seemed rare at weeks 52. PMID- 17400305 TI - Dissociable learning-dependent changes in REM and non-REM sleep in declarative and procedural memory systems. AB - Sleep spindles and rapid eye movements have been found to increase following an intense period of learning on a combination of procedural memory tasks. It is not clear whether these changes are task specific, or the result of learning in general. The current study investigated changes in spindles, rapid eye movements, K-complexes and EEG spectral power following learning in good sleepers randomly assigned to one of four learning conditions: Pursuit Rotor (n=9), Mirror Tracing (n=9), Paired Associates (n=9), and non-learning controls (n=9). Following Pursuit Rotor learning, there was an increase in the duration of Stage 2 sleep, spindle density (number of spindles/min), average spindle duration, and an increase in low frequency sigma power (12-14Hz) at occipital regions during SWS and at frontal regions during Stage 2 sleep in the second half of the night. These findings are consistent with previous findings that Pursuit Rotor learning is consolidated during Stage 2 sleep, and provide additional data to suggest that spindles across all non-REM stages may be a mechanism for brain plasticity. Following Paired Associates learning, theta power increased significantly at central regions during REM sleep. This study provides the first evidence that REM sleep theta activity is involved in declarative memory consolidation. Together, these findings support the hypothesis that brain plasticity during sleep does not involve a unitary process; that is, different types of learning have unique sleep related memory consolidation mechanisms that act in dissociable brain regions at different times throughout the night. PMID- 17400304 TI - Cycloheximide: no ordinary bitter stimulus. AB - Cycloheximide (CyX), a toxic antibiotic with a unique chemical structure generated by the actinomycete, Streptomyces griseus, has emerged as a primary focus of studies on mammalian bitter taste. Rats and mice avoid it at concentrations well below the thresholds for most bitter stimuli and T2R G protein-coupled receptors specific for CyX with appropriate sensitivity are identified for those species. Like mouse and rat, golden hamsters, Mesocricetus auratus, also detected and rejected micromolar levels of CyX, although 1mM CyX failed to activate the hamster chorda tympani nerve. Hamsters showed an initial tolerance for 500microM CyX, but after that, avoidance of CyX dramatically increased, plasticity not reported for rat or mouse. As the hamster lineage branches well before division of the mouse-rat lineage in evolutionary time, differences between hamster and mouse-rat reactions to CyX are not surprising. Furthermore, unlike hamster LiCl-induced learned aversions, the induced CyX aversion neither specifically nor robustly generalized to other non-ionic bitter stimuli; and unlike adverse reactions to other chemosensory stimuli, aversions to CyX were not mollified by adding a sweetener. Thus, CyX is unlike other bitter stimuli. The gene for the high-affinity CyX receptor is a member of a cluster of five orthologous T2R genes that are likely rodent-specific; this "CyX clade" is found in the mouse, rat and probably hamster, but not in the human or rabbit genome. The rodent CyX-T2R interaction may be one of multiple lineage-specific stimulus-receptor interactions reflecting a response to a particular environmental toxin. The combination of T2R multiplicity, species divergence and gene duplication results in diverse ligands for multiple species-specific T2R receptors, which confounds definition of 'bitter' stimuli across species. PMID- 17400306 TI - Psychomotor vigilance task performance during total sleep deprivation in young and postmenopausal women. AB - The objective of this study was to investigate the effects of age on women's performance in the psychomotor vigilance task (PVT) during total sleep deprivation (SD). A total of 46 healthy women volunteered. They belonged to two age groups: young (n=34; age range 19-30 years; 12 without, and 22 with oral contraceptives (OC); early phase of the menstrual cycle) and older (n=12; age range 60-68; postmenopausal; without hormone therapy). During a 40-h total SD, the subjects performed the PVT and the Stanford Sleepiness Scale (SSS) at 2-h intervals. At baseline, the reaction speed of the young women was faster as compared to the older women (Mann-Whitney U-test p<0.01). During SD, all the PVT measures as well as the SSS scores changed similarly in the two age groups, when the baseline performance difference in favour of the young women was taken into account (area under curve analyses, Mann-Whitney U-tests n.s.). No age difference in the time course of the SD-related deterioration in PVT performance or subjective sleepiness was observed. OC use had no effects on any of the measures during SD. After recovery sleep, young women had higher subjective sleepiness scores than older women, the sleepiness scores being highest in young women not taking OCs. In conclusion, in women, aging has no effects on the amount or the time course of the decline in PVT performance caused by total SD. OC use does not significantly affect young women's PVT performance during SD in the early phase of the menstrual cycle. PMID- 17400307 TI - Relocalization of the polypyrimidine tract-binding protein during PKA-induced neurite growth. AB - Neurite RNA binding proteins are important for neurite growth, a process critical for neuronal development and regeneration after injury. It has been known that many RNA binding proteins undergo nucleocytoplasmic shuttling but how their nucleocytoplasmic distributions are regulated during neurite growth has not been well explored. Here we found that the polypyrimidine tract binding protein (PTB) was exported from the nucleus and accumulated at growing neurite terminals upon activation of the PKA pathway in PC12 cells in a PKA-target Ser16-dependent manner. RNA interference (RNAi) of PTB significantly disrupted the neurite growth. We then examined the role of cytoplasmic PTB in relation to mRNAs involved in neurite growth. We found that PTB was preferentially associated with the beta-actin mRNA transcripts in cytoplasmic fractions. RNAi of PTB reduced neurite accumulation of the endogenous actin proteins. It is thus likely that, during PKA-induced neurite growth, PTB is relocalized through Ser16 phosphorylation to the cytoplasm where it is associated with beta-actin mRNA and is critical for the mRNA localization to neurites. PMID- 17400308 TI - Simultaneous hemodynamic and serological cardiotoxicity monitoring during immunotherapy with trastuzumab. AB - BACKGROUND: Immunotherapy with trastuzumab (Herceptin), a selective HER-2(ErbB2) antibody, is associated with a certain degree of cardiotoxicity. This study sought to evaluate the immediate hemodynamic response to trastuzumab with real time CW-Doppler depending on the level of nt-pro-BNP (brain natriuretic peptide) as a possible marker of cardiotoxicity. METHODS: 48 patients with HER-2-positive metastatic breast cancer were continuously measured with CW-Doppler ultrasound for cardiac output (CO) and systemic vascular resistance (SVR) before, during and after drug infusion in combination with nt-pro-BNP before and 10 min after drug infusion. Depending on the nt-pro-BNP-levels <125 pg/ml (group A, n=34, 51+/-11 years) vs. nt-pro-BNP >125 pg/ml (group B, n=14, 63+/-7 years) two groups have been defined. RESULTS: Trastuzumab therapy did not change nt-pro-BNP immediately before (44+/-29 pg/ml) vs. after the infusion (45+/-32 pg/ml, n.s.) in the low level as in the high level nt-pro-BNP group (231+/-356 pg/ml prior and 240+/-377 pg/ml, n.s.). Cardiac output remained stable during trastuzumab infusion, however cardiac output was significantly increased following the end of the infusion stronger in the high-level nt-pro-BNP group. Systemic vascular resistance prior to the trastuzumab infusion was higher in the high-level nt-pro-BNP group with significant decrement during and after the infusion. CONCLUSION: Combining real time CW-Doppler ultrasound and nt-pro-BNP monitoring is feasible to monitor the immediate hemodynamic changes during and after trastuzumab infusion. PMID- 17400309 TI - Regression of paroxysmal atrial fibrillation associated with thiazolidinedione therapy. AB - Thiazolidinediones (TZDs) represent insulin-sensitizing agents that have several pleiotropic properties, possibly related to their favorable effects on cardiovascular remodeling. We briefly describe 2 diabetic patients who experienced a remarkable improvement in their paroxysmal atrial fibrillation (AF) after treatment with rosiglitazone. Current evidence suggests that atrial remodeling represents a prominent mechanism of AF development and perpetuation while inflammation and oxidative stress are possibly implicated in this process. It could therefore be speculated that the pleiotropic effects of TZDs favorably affect atrial remodeling reducing the arrhythmia burden. Further studies are needed in order to elucidate the merit of this pharmacological approach in AF. PMID- 17400310 TI - Comment on the article by Vazquez and Marquez-Garcia. High-risk vs. low-risk for thrombo-embolic events: who are in the intermediate-risk group? What should be done then? AB - Although high risk and low risk profile for thrombo-embolic events have been extensively and intensively investigated in multi-centre trials and described in detail in atrial fibrillation guidelines, the management of those at otherwise intermediate risk is still a 'no-one's zone'. Individual risk profile is mandatory to identify those who will benefit of anti-coagulation therapy and large-scale clinical trials are still awaited to define efficacy and efficiency profile in intermediate-risk group. PMID- 17400311 TI - Comparisons of short- and intermediate-term effects of pitavastatin versus atorvastatin on lipid profiles, fibrinolytic parameter, and endothelial function. AB - We compared short- and intermediate-term effects on lipid profiles, fibrinolytic parameter, and endothelial function between pitavastatin and atorvastatin. Short term improvement of endothelial function was superior with pitavastatin compared to atorvastatin therapy. Pitavastatin could be a potentially better therapeutic choice for lipid-lowering and early alterations in endothelial function. Our study provides an important basis on which further trials involving larger numbers of patients may be studied prospectively. PMID- 17400312 TI - A segmental approach to criss-cross heart by cardiac MRI. AB - Criss-cross heart is a rare congenital heart defect. The unusual arrangement of the atrio-ventricular connection, and associated abnormalities make it a challenging condition to image. We describe 3 cases of criss-cross heart who underwent cardiac MRI in order to comprehensively evaluate the anatomy and plan surgical management. PMID- 17400313 TI - The enigma of quality of life in patients with heart failure. AB - Current treatment goals in heart failure (HF) aim to improve both survival and quality of life (QoL) of patients. In this brief communication, we reviewed randomized controlled trials that assessed the impact of pharmacological treatment on QoL, and we discussed some methodological limitations of QoL assessment in HF. Studies that assessed QoL with a disease-specific questionnaire were included. We found that at present there is a paradox in HF treatment. Life prolonging therapies, such as angiotensin-converting-enzyme-inhibitors, and angiotensin receptor blockers improve modestly or only delay the progressive worsening of QoL in HF. Treatment with beta blockers does not affect QoL in any way. However, this neutral effect of beta blockers may also be due to some methodological limitations, such as the small number of patients included in beta blocker trials or the short duration of follow-up. Disease-specific questionnaires may also have some limitations, e.g. are not sensitive enough to detect small changes in QoL. On the other hand, therapies that significantly improve QoL in HF (e.g. inotropic agents) do not seem beneficial in relation to survival. We conclude that QoL in HF remains an open field, in which new therapies but also clarification of methodology is required. In the mean time, the use of life prolonging therapies appears as a safe measure to modestly improve or maintain QoL. PMID- 17400314 TI - Sotalol unmasks susceptibility to drug-induced long QT syndrome and torsades de pointes. PMID- 17400315 TI - Buddy wire technique for stent placement at non-aorto ostial coronary lesions. AB - Percutaneous coronary intervention (PCI) of coronary non-aorto ostial lesions offers technical challenges not encountered with other lesion sub types. Stenting of these lesions improves acute angiographic result and lowers the rate of restenosis. However, precise stent placement at non-aorto ostial lesions is technically difficult with risk of incomplete lesion coverage or jailing of the main branch. In this report, we describe a buddy wire technique to facilitate precise stent placement at non-aorto ostial lesions. PMID- 17400316 TI - Selection of cases and controls: Additional information is needed. AB - The potential for selection bias is a concern in any case-control study. Cases and controls should be representative of the respective diseased and nondiseased persons in a well delineated target population. Even with representative cases and controls, if there is a high (or differential) nonparticipation rate among (or between) cases and controls, selection bias may be introduced. Case-control studies offer an efficient observational analytic design, but they also carry a high potential for the introduction of bias. PMID- 17400317 TI - Electrophysiological indices of processing aesthetics: Spontaneous or intentional processes? AB - Processes underlying aesthetic appreciation of formal graphic black and white patterns were investigated. In previous EEG studies on aesthetic and symmetry judgments, an early frontocentral negativity has been observed for not beautiful judgments, and interpreted as reflecting an impression formation. In addition, a lateralized late positivity, more pronounced in the aesthetic than in the symmetry judgment task, has been interpreted as reflecting evaluative categorization; and a sustained posterior negativity for not symmetric judgments, has been interpreted as reflecting a prolonged symmetry analysis. Here, we investigated whether these processes occur spontaneously, i.e. independent of an aesthetics task instruction or a judgment task demand. Participants were randomly assigned to two group conditions. In the Viewing condition, participants were instructed to view graphic patterns, and to detect a probe. In the Contemplation condition, participants were instructed to contemplate the beauty of the patterns, and to detect a probe. No aesthetics-related response was required in either condition. ERP results suggest that symmetry analysis occurred spontaneously. Evaluative categorization, reflected by a late positivity, required an aesthetics instruction and did not occur spontaneously. This process was elicited without an overt aesthetic judgment. An early frontocentral negativity for not beautiful patterns, interpreted as reflecting impression formation, did not occur (in contrast to previous studies that required an overt aesthetic judgment). Given the present data, aesthetic appreciation of graphic patterns requires different sub-processes. Aesthetic contemplation is reflected by a lateralized late positivity, whilst an aesthetic judgment is additionally reflected by an early frontocentral negativity. Aesthetic appreciation of beauty appears to require intention and is not spontaneous in character. PMID- 17400318 TI - Environmental analysis of plastic production processes: comparing petroleum-based polypropylene and polyethylene with biologically-based poly-beta-hydroxybutyric acid using life cycle analysis. AB - Polymers based on olefins have wide commercial applicability. However, they are made from non-renewable resources and are characterised by difficulty in disposal where recycle and re-use is not feasible. Poly-beta-hydroxybutyric acid (PHB) provides one example of a polymer made from renewable resources. Before motivating its widespread use, the advantages of a renewable polymer must be weighed against the environmental aspects of its production. Previous studies relating the environmental impacts of petroleum-based and bio-plastics have centred on the impact categories of global warming and fossil fuel depletion. Cradle-to-grave studies report equivalent or reduced global warming impacts, in comparison to equivalent polyolefin processes. This stems from a perceived CO(2) neutral status of the renewable resource. Indeed, no previous work has reported the results of a life cycle assessment (LCA) giving the environmental impacts in all major categories. This study investigates a cradle-to-gate LCA of PHB production taking into account net CO(2) generation and all major impact categories. It compares the findings with similar studies of polypropylene (PP) and polyethylene (PE). It is found that, in all of the life cycle categories, PHB is superior to PP. Energy requirements are slightly lower than previously observed and significantly lower than those for polyolefin production. PE impacts are lower than PHB values in acidification and eutrophication. PMID- 17400319 TI - Systems biology--an engineering perspective. AB - The interdisciplinary field of systems biology has evolved rapidly over the last years. Different disciplines have aided the development of both its experimental and theoretical branches. One field, which has played a significant role is engineering science and, in particular chemical engineering. Here, we review and illustrate some of these contributions, ranging from modeling approaches to model analysis with a special focus on technique which have not yet been substantially exploited but can be potentially useful in the analysis of biochemical systems. PMID- 17400320 TI - 10-Hydroxycamptothecin loaded nanoparticles: preparation and antitumor activity in mice. AB - 10-Hydroxycamptothecin (HCPT) loaded nanoparticles made from poly(caprolactone-co lactide)-b-PEG-b-poly(caprolactone-co-lactide) (PCLLA-PEG-PCLLA) block copolymer, were prepared by a novel two-step nanoprecipitation method using an interior chemistry strategy. The satisfactory drug loading content (>13%) as well as high encapsulation efficiency (>85%) was achieved. Cytotoxicity test indicated that the HCPT-loaded nanoparticles had enhanced in vitro cytotoxicity compared to free drug. Progressively, in vivo antitumor activity and HCPT biodistribution in sarcoma-180 (S-180) bearing mice after intravenous injection of the HCPT-loaded nanoparticles show that HCPT-loaded nanoparticles exhibited superior in vivo antitumor effect and remarkably different biodistribution than the commercially available HCPT injection. PMID- 17400322 TI - The natural history of hepatitis C with severe hepatic fibrosis. AB - BACKGROUND/AIMS: To examine the morbidity and mortality of patients with severe fibrosis secondary to HCV infection, within a population unbiased by tertiary referral. METHODS: One hundred and fifty HCV infected patients were identified from the Trent HCV study with a liver biopsy taken before 2002 demonstrating severe fibrosis (Ishak stage > or =4). Follow-up data were extracted from the database and hospital records. RESULTS: Median follow-up was 51 months. Of the 131 patients with no prior history of decompensation, 33 (25%) died (n=25) or were transplanted (n=8), after a median interval of 42 months. The probability of survival without liver transplantation was 97%, 88%, and 78% at 1, 3, and 5 years, respectively. Hepatocellular carcinoma and/or decompensation was diagnosed in 33 (25%), after a median interval of 41 months. In multivariate analysis, combination antiviral therapy was associated with improved survival. Prognosis was not affected by the Ishak stage at index biopsy. There was a worse prognosis for the 19 patients with previous decompensation; 17 (89%) having either died (n=15) or been transplanted (n=2). CONCLUSIONS: This study demonstrates that severe liver fibrosis (Ishak stage > or = 4) secondary to hepatitis C is associated with a poor prognosis, that may be improved following combination antiviral treatment. PMID- 17400323 TI - Prevalence and risk factors of non-alcoholic fatty liver disease in potential living liver donors in Korea: a review of 589 consecutive liver biopsies in a single center. AB - BACKGROUND/AIMS: The aim of the study was to evaluate the prevalence and risk factors of biopsy-proven non-alcoholic fatty liver disease (NAFLD) in potential living liver donors and to evaluate the efficacy of imaging techniques for the detection of steatosis in donors. METHODS: We reviewed the results of liver biopsy, ultrasonography (USG) and computed tomography (CT) and biochemical data performed in 589 consecutive potential liver donors as a pretransplantation work up from July 2004 to September 2005 at Asan Medical Centre. RESULTS: Of 589 participants, 408 (69.3%) were men, with a mean age of 31.1+/-9.5 years. NAFLD (5% steatosis in biopsy) was diagnosed in 303 (51.4%); >30% steatosis in 61 (10.4%) and non-alcoholic steatohepatitis in 13 (2.2%). The independent risk factors for >30% steatosis were age over 30 (OR=2.223; p=0.014), obesity (OR=5.320; p<0.001) and hypertriglyceridemia (OR=2.253; p=0.019) by multivariate analysis. The sensitivity of USG and CT for detecting >30% steatosis was 92.3% and 64.1%, and positive predictive value was only 34.5% and 45.1%, respectively. CONCLUSIONS: NAFLD was highly prevalent in potential living liver donors. The independent risk factors for significant steatosis were older age, obesity and hypertriglyceridemia. USG and CT had limitations in detecting significant steatosis in liver donors. PMID- 17400324 TI - Genetic polymorphisms of manganese superoxide dismutase, NAD(P)H:quinone oxidoreductase, glutathione S-transferase M1 and T1, and the susceptibility to drug-induced liver injury. AB - BACKGROUND/AIMS: Drug metabolizing enzymes may be related to drug-induced liver injury (DILI). Manganese superoxide dismutase (MnSOD), NAD(P)H:quinone oxidoreductase (NQO1), and glutathione S-transferase (GST) are important drug metabolizing enzymes. We aimed to elucidate the relationship between genetic polymorphisms of these enzymes and the susceptibility to DILI. METHODS: A total of 115 patients with DILI and 115 drug-, sex-, and age-matched controls were enrolled. Their genetic polymorphisms of MnSOD, NQO1, GSTM1, and GSTT1 were assayed. RESULTS: Sixty-three (54.8%) of DILI patients were incriminated to anti tuberculosis drugs. Subjects with a mutant C allele (T/C or C/C genotype) of MnSOD had a higher risk of DILI than those with MnSOD T/T genotype, both in overall drugs studied (adjusted OR: 2.44, 95% C.I.: 1.38-4.30, P=0.002), and in sub-category of anti-tuberculosis drugs (adjusted OR: 2.47, 95% C.I.: 1.13-5.39, P=0.02). In addition, subjects carrying GSTM1 null genotype had increased risk of anti-tuberculosis DILI (adjusted OR: 2.23, 95% C.I.: 1.07-4.67, P=0.03). CONCLUSIONS: The MnSOD mutant C allele may increase the susceptibility to DILI, and GSTM1 null genotype may be related to anti-tuberculosis drug-induced hepatotoxicity. Determination of the MnSOD and GSTM1 genotypes may help identify patients at high risk for DILI. PMID- 17400325 TI - Statins in non-alcoholic fatty liver disease and chronically elevated liver enzymes: a histopathological follow-up study. AB - BACKGROUND/AIMS: The effect of statins on hepatic histology in non-alcoholic fatty liver disease (NAFLD) is not known. This study explores hepatic histology in NAFLD patients before and after initiation of statin therapy and compares histological outcome with NAFLD patients who had not been prescribed statins. METHODS: Sixty-eight NAFLD patients were re-evaluated. Follow-up ranged from 10.3 to 16.3 years. Subjects were clinically investigated and a repeat liver biopsy was obtained. No patient was taking statins at baseline while 17 patients were treated with statins at follow-up. RESULTS: At baseline, patients that later were prescribed statins had significantly higher BMI and more pronounced hepatic steatosis. At follow-up patients on medication with statins continued to have significantly higher BMI. Diabetes was significantly more common among patients on medication with statins and they had significantly more pronounced insulin resistance. However, they exhibited a significant reduction of liver steatosis at follow-up as opposed to patients not taking statins. Despite exhibiting a high risk profile for progression of liver fibrosis, only four patients on statin treatment progressed in fibrosis stage. CONCLUSIONS: Statins can be prescribed in patients with elevated liver enzymes because of NAFLD. PMID- 17400326 TI - Bradykinesia in cirrhotic patients with early hepatic encephalopathy is related to a decreased glucose uptake of frontomesial cortical areas relevant for movement initiation. AB - BACKGROUND/AIMS: Bradykinesia is one of the first symptoms of hepatic encephalopathy (HE). Recently it has been suggested that bradykinesia in HE is due to disturbances in movement initiation. Areas involved in self-initiated movement are the motor- and premotor cortex, the supplementary motor cortex, the motor areas of the cingulate gyrus, and part of the frontomesial- and parietal cortex. The present study aimed to test the hypothesis that bradykinesia in HE is due to a functional disturbance of these areas. METHODS: Fourteen cirrhotics with grade 0-I HE were examined. Patients with alcoholic cirrhosis or concomitant cerebral disorder were excluded. Patients underwent a 3-dimensional computer assisted movement analysis for forearm pronation and supination, hand tapping and finger tapping and a (18)F-fluorodesoxy-glucose-PET-examination during rest, analysed with statistical parametric mapping (SPM99). RESULTS: The frequency of finger- and hand tapping was significantly correlated to the glucose metabolism of the motor area of the cingulate gyrus and frontomedial, frontodorsal and parietal cortical areas known to be activated with self-initiated movements. A decrease of movement frequency was associated with a reduction of glucose metabolism within these areas. CONCLUSIONS: These data support the hypothesis that bradykinesia in cirrhotics with HE is caused by an alteration of movement initiation. PMID- 17400327 TI - Are chromosomal imbalances important in cancer? AB - Tumor-specific patterns of large-scale chromosomal imbalances characterize most forms of cancer. Based on evidence primarily from neuroblastomas, it can be argued that large-scale chromosomal imbalances are crucial for tumor pathogenesis and have an impact on the global transcriptional profile of cancer cells, and that some imbalances even initiate cancer. The genes and genetic pathways that have been dysregulated by such imbalances remain surprisingly elusive. Many genes are affected by the regions of gain and loss, and there are complex interactions and relationships that occur between these genes, hindering their identification. The study of untranslated RNA sequences, such as microRNAs, is in its infancy, and it is likely that such sequences are also dysregulated by chromosomal imbalance, contributing to pathogenesis. PMID- 17400328 TI - Scientific production of electronic health record research, 1991-2005. AB - PURPOSE: The increasing numbers of publications on electronic health record (EHR) indicate its increasing importance in the world. This study attempted to quantify the scientific production of EHR research articles, and how they have changed over time, in an effort to investigate changes in the trends cited in these critical evaluations. METHOD: The articles were based on the science citation index (SCI) from 1991 to 2005. A descriptive study was performed using the 1803 documents published in the SCI from 39 countries in America, Europe, Africa, Asia, and Oceania. The evaluation was based on parameters including document type, language, first author's country of origin, number of citations and citations per publication. RESULTS: Of all publications, 1455 (80.7%) were articles, followed by meeting abstracts which represented about one-tenth of all types of EHR publications. Numbers of published articles have significantly increased when compared by each 5-year period. Most articles were published in English (98%) and were from the region of America (57%). The top 10 of the 374 journals accounted for 41% of the number of published articles. The US dominates publication production (57%) with a cumulative impact factor (IF) of 2227 and followed by the UK (8.5%, with a cumulative IF of 257.0) and the Netherlands (7.8%, with a cumulative IF of 211.1). An analysis of the number of articles related to population revealed a high publication output for relative small countries like Switzerland, the Netherlands, and Norway. CONCLUSIONS: Research production in EHR showed a considerable increase during 1991-2005. The production was dominated by articles, those from the US, and those published in English. The production came from many countries, denoting the devotion to this field in different areas around the world. PMID- 17400329 TI - Pneumocystis pneumonia in a patient with non-small cell lung cancer (NSCLC) treated with pemetrexed containing regimen. AB - Pneumocystis pneumonia is typically life-threatening in immunocompromised patients. It is relatively uncommon in patients with lung cancer. We report a case of pneumocystis pneumonia in a patient with advanced stage non-small cell lung cancer (NSCLC) following treatment with concurrent chemoradiation with a pemetrexed containing regimen. To our knowledge, this is the first report on pneumocystis pneumonia following administration of pemetrexed containing regimen. PMID- 17400330 TI - Malignant pleural mesothelioma: two cases in first degree relatives. AB - We report two cases of malignant pleural mesothelioma in first degree relatives arising within weeks of each other. The patients had a shared exposure to asbestos and age difference of over 20 years at time of presentation. In both cases, the anatomical pattern was similar and unusual for mesothelioma and initial histology was reported for both as non-small cell lung cancer. Both patients were treated with platinum-based combination chemotherapy. PMID- 17400331 TI - Detection of oncogenic viruses SV40, BKV, JCV, HCMV, HPV and p53 codon 72 polymorphism in lung carcinoma. AB - As a part of our continuous search for oncogenic viruses in bronchial cancer, we extended our HPV studies to analyse also SV40, BKV, JCV and HCMV sequences in bronchial cancer and related these data with p53 codon 72 polymorphism. Fresh tumor samples from 78 patients with lung cancer were analysed for SV40, BKV, JCV, HCMV and HPV sequences by PCR. HPV genotypes were determined using reverse blot hybridization and sequencing, and all HPV-positive tumors were tested for the presence of E6/E7 transcripts by RT-PCR. All samples were analysed for p53 codon 72 polymorphism, using PCR-based RFLP method. Of the 78 cases studied, 11 (14.1%) were positive for T-Ag gene of SV40, while BKV and JCV sequences were both amplified in 1 tumor only. Altogether, 10/78 lesions were HPV-positive; six HPV16, one HPV31, two HPV6/53 and one HPV16/18. All HPV DNA-positive samples except one also expressed E6 and E7 transcripts. HCMV was amplified in 18 (23%) cases. RFLP analysis of p53 codon 72 revealed 32 homozygotes for arg/arg allele (50.8%), 26 heterozygotes for arg/pro allele (41.3%), and 5 homozygotes for pro/pro allele (7.9%). P53 codon 72 polymorphism was not significantly different between cases (n=63) and controls (n=50) (p=0.455), among virus positive and negative patients, nor was it related to HPV genotypes (p=0.384), expression of E6 (p=0.384) and E7 oncogenes (p=0.293). Of all possible combinations of virus co detection, only SV40-HCMV association was statistically significant (OR=5.500, 95%CI 1.43-21.02; p=0.015). Taken the known mechanisms of these individual viruses, there is a chance that these viruses could affect cell cycle control and inhibit apoptosis, thus potentially causing genetic instability and promote oncogenesis. PMID- 17400332 TI - Lack of evidence that p53 Arg72Pro influences lung cancer prognosis: an analysis of survival in 619 female patients. AB - The prognostic significance of the Arg72Pro polymorphism of the p53 tumour suppressor gene in cancer is controversial. To determine whether Arg72Pro is a marker for lung cancer prognosis we genotyped 619 female lung cancer patients with incident disease and examined the relationship between genotype and overall survival (OS). Nonparametric tests provided no evidence for a relationship between SNP genotype and OS (P-values 0.131, 0.161, and 0.156 for log rank, Wilcoxon and Fleming-Harrington test statistics, respectively). Under the Cox proportional hazards model the HRs associated with Arg/Pro, Pro/Pro and Pro carrier status were: 0.98 (95%CI: 0.79-1.22), 0.76 (95%CI: 0.51-1.15) and 0.93 (95%CI: 0.76-1.15), respectively. Despite employing a comprehensive set of statistical tests including those sensitive to the detection of differences in early survival our data provide little evidence to support the tenet that the p53 Arg72Pro polymorphism is a clinically useful prognostic marker for lung cancer. PMID- 17400333 TI - Development of photolabile caged analogs of endothelin-1. AB - Photoactivable caged analogs of endothelin-1 (ET-1) were obtained after derivatization with the photolabile 4,5-dimethoxynitrobenzyl (DMNB) group. This was achieved by the incorporation of N-alpha-Fmoc caged building blocks of Lys, Asp, Glu and Tyr during the solid phase peptide synthesis step. The C-terminal carboxylic function was also derivatized. However, difficulties were encountered with the introduction of the Asp and Glu photoactivable building blocks. As a matter of fact, formation of an aminosuccinyl derivative, through cyclization of the Asp(ODMNB) residue, and the formation of a pyrrolidone ring from the Glu(ODMNB) residue were highly favored by the electronic properties of the photocleavable function. ET-1 analogs were also tested in the ET(A) and ET(B) paradigms and specific pharmacological profiles were obtained for each peptide. PMID- 17400334 TI - Genetic loci modulating amyloid-beta levels in a mouse model of Alzheimer's disease. AB - Genetic studies have demonstrated very high heritability for Alzheimer's disease (AD) risk in humans; however, these genetic contributions have proven extremely challenging to map in large studies of AD patients. Processing of the amyloid precursor protein (APP) to produce amyloid-beta (Abeta) peptide is increasingly believed to be of central importance in AD pathogenesis. Intriguingly, mice from the C57BL/6J and DBA2/J inbred strains carrying the R1.40 APP transgene produce identical levels of unprocessed APP, but demonstrate significant, heritable differences in Abeta levels. To identify specific loci responsible for the observed genetic control of Abeta metabolism in this model system, we have performed a whole-genome quantitative trait locus (QTL) mapping experiment on a total of 516 animals from a C57BL/6JxDBA/2J intercross using a dense set of SNP genetic markers. Our studies have identified three loci on mouse chromosomes 1, 2, and 7 showing significant or suggestive associations with brain Abeta levels, several of which contain regions syntenic to previous reports of linkage in human AD. PMID- 17400335 TI - Using practitioner research to investigate the role of the teacher in encouraging student interaction within group work. AB - This paper promotes the use of self-reflective enquiry through practitioner research to address the research question How can I encourage more equal participation of students in group discussions? The research is located in two consecutive continuing professional development modules for nurses caring for older people. The development of the research question is tracked through student evaluation of the modules. Background theory is presented in relation to both student interaction and the teacher's role in group learning and the method of enquiry, practitioner research. Two action cycles are undertaken and described in which a critical friend is asked to record student interaction with the use of a sociogram. The findings identify that controlling group membership can lead to a greater perceived ability to participate in group discussions. Techniques to encourage student-student interaction are identified and it is suggested that positive benefits of learning result. The lessons learnt and areas for further research are considered. PMID- 17400336 TI - Nursing education in the Czech Republic. AB - The report is focused on the transformation process in pregraduate education of nurses in the Czech Republic. The all-society changes taking place in the last decades of the 20th century have opened the way for implementation of changes in the Czech nursing as well. Nursing is becoming an independent scientific discipline, the role of the nurses and their position in the society are changing and the professional preparation is gradually shifting to colleges and universities. After 15 years of effort to implement the planned transformation changes in nursing education, the Czech Republic's joining of the European Union has helped us. In connection with fulfillment of Bologna declaration, Master study in nursing has been gradually developing in education of nurses in the Czech Republic as well. In order to create a structured study in nursing according to the above mentioned declaration, a postgraduate doctor program must be implemented too. The implementation of all types of university education of nurses is very important and it will positively contribute to the development of nursing in our republic as a full-bodied scientific discipline and to the creation of its high-quality scientific-research base. PMID- 17400337 TI - Antibody titer in Iranian children 6 years after hepatitis B vaccine administration. AB - OBJECTIVE: The purpose of study was to evaluate the immune response in a sample of vaccinated children aged 6 years. BACKGROUND: Although immunization of infants against hepatitis B virus (HBV) is the most effective way to prevent infection, duration of the afforded immunization is unknown. METHODS: The immunity derived from the HBV vaccine was assessed by measuring the antibody in 3752 children who were vaccinated in a routine vaccination program in three cities of Iran (Isfahan, Khoramabad, Shahrekord). RESULTS: Seven hundred and twenty-three (19.3%) children had antibodies levels <10 MIU/mL and 1096 (29.2%) had antibodies levels >or=100 MIU/mL. The total GMT was 34.5+/-0.66, and GMT was statistically different in non-immune and immune children (3.1+/-0.36 versus 49.1+/-0.52). No correlation was found between HbsAb titers and growth pattern during the first and sixth years of life, number of vaccine, time of vaccination and drug use. The predictors were low birth weight and chronic disease. CONCLUSION: It is recommended that high risk children should be monitored regularly for anti-HBS, and booster must be administrated, if necessary. PMID- 17400338 TI - Pneumococcal vaccine in Italy. PMID- 17400339 TI - The development of standardized case definitions and guidelines for adverse events following immunization. PMID- 17400340 TI - Fatigue: case definition and guidelines for collection, analysis, and presentation of immunization safety data. PMID- 17400341 TI - Evaluating rotavirus vaccination in England and Wales. Part II. The potential cost-effectiveness of vaccination. AB - AIM: Rotavirus is the leading cause of acute gastroenteritis in children. Two rotavirus vaccines (RotaTeq and Rotarix) have recently completed clinical trials. We investigated whether routine infant immunisation with either vaccine can be cost effective. METHODS: We compared costs and outcomes of vaccination using a cohort model, following children over the first 5 years of life. We estimated health provider costs, economic costs and quality adjusted life years (QALYs) lost due to rotavirus-related deaths, hospital admissions, nosocomial infections, accident and emergency attendances, general practice consultations and calls to NHS Direct. RESULTS: Under base case assumptions, a programme using RotaTeq (priced at pound 25 a dose) would cost the health provider pound 79,900 per QALY gained. Using Rotarix (priced at pound 35 a dose) would cost pound 61,000 per QALY gained. Univariate and multivariate sensitivity analysis indicate that at these prices an immunisation programme would be unlikely to be cost-effective for any realistic value of the key parameters. CONCLUSIONS: Rotavirus immunisation could reduce the substantial short-term morbidity burden due to rotavirus, but is unlikely to be deemed cost effective unless the vaccine is competitively priced. PMID- 17400342 TI - Do immunisations reduce the risk for SIDS? A meta-analysis. AB - BACKGROUND: There are claims that immunisations cause sudden infant death syndrome (SIDS), but some studies have found either no association or that they are associated with a reduced risk of SIDS. AIMS: To conduct a meta-analysis examining the relationship between immunisation and SIDS. METHODS: Nine case controls studies were identified examining this association, of which four adjusted for potential confounders. RESULTS: The summary odds ratio (OR) in the univariate analysis suggested that immunisations were protective, but the presence of heterogeneity makes it difficult to combine these studies. The summary OR for the studies reporting multivariate ORs was 0.54 (95% CI=0.39-0.76) with no evidence of heterogeneity. CONCLUSIONS: Immunisations are associated with a halving of the risk of SIDS. There are biological reasons why this association may be causal, but other factors, such as the healthy vaccinee effect, may be important. Immunisations should be part of the SIDS prevention campaigns. PMID- 17400343 TI - Observation of changes in urinary excretion of thorium in humans following ingestion of a therapeutic soil. AB - The study investigated the changes in urinary thorium excretion by humans following ingestion of a therapeutic soil, which contains about 10 ppm of thorium. This well-known healing earth in Germany has been considered as an alternative medicine for diarrhoea and gastric hyper-acidity. Six adult volunteers ingested this therapeutic soil in varying quantities for 1-15 days at levels approximating those described in the package insert of the medicine (10-60 g of soil per day). The subjects ingested about 0.1-0.6 mg of thorium daily, which is 100-600 times higher than the normal daily intake of about 1 microg thorium in Germany. All 24-h urine samples collected from the subjects during pre ingestion, ingestion and post-ingestion periods of the soil were analyzed for (232)Th using inductively coupled plasma mass spectrometry (ICP-MS). The measured excretion values varied in a wide range. Apparently, the high thorium amounts administered did not increase the (232)Th excretion in urine as expected, suggesting that this soil ingestion will not result in a considerably higher and harmful uptake of thorium into the human body. PMID- 17400344 TI - Radiostrontium hot spot in the Russian Arctic: ground surface contamination by (90)Sr at the "Kraton-3" underground nuclear explosion site. AB - Strontium-90 activity concentrations in surface soils and areal deposition densities have been studied at a site contaminated by an accidental release to atmosphere from the underground nuclear explosion "Kraton-3" conducted near the Polar Circle (65.9 degrees N, 112.3 degrees E) within the territory of the former USSR in 1978. In 2001-2002, the ground surface contamination at 14 plots studied ranged from 20 to 15 000 kBq m(-2), which significantly exceeds the value of 0.44 kBq m(-2) deduced for three background plots. The zone with substantial radiostrontium contamination extends, at least, 2.5 km in a north-easterly direction from the borehole. The average (137)Cs/(90)Sr ratio in the ground contamination originated from the "Kraton-3" fallout was estimated to be 0.55, which is significantly different from the ratio of 2.05 evaluated for background plots contaminated mostly from global fallout. Although vertical migration of (90)Sr in all undisturbed soil profiles studied is more rapid than that for (137)Cs, the depth of percolation of both radionuclides into the ground is mostly limited to the top 10-20 cm, which may be explained, primarily, by permafrost conditions. The horizontal migration rate of radiostrontium in the aqueous phase exceeds the radiocaesium migration rate by many times. This phenomenon seems to be a reason for the significant enrichment of the soil surface layers by radiostrontium at some sites, with variations occurring in accordance with small scale irregularities of landscape. PMID- 17400345 TI - Natural radioactivity in some major rivers of coastal Karnataka on the southwest coast of India. AB - Systematic studies on radiation level and distribution of radionuclides have been carried out in riverine environs of three major rivers of coastal Karnataka, viz. Kali, Sharavathi and Netravathi. The ambient gamma radiation levels along three rivers were measured using a portable plastic scintillometer. Activity concentrations of (226)Ra, (232)Th and (40)K in soil, sediment and rock were measured using a NaI(Tl) gamma-ray spectrometer. In the Kali, Sharavathi and Netravathi riverbanks, the median values of absorbed gamma dose rates in air were found to be 44 nGy h(-1), 35 nGy h(-1) and 57 nGy h(-1), respectively. The highest activity of (226)Ra was found in riverbank soil samples of Sharavathi River. The highest activities of (232)Th and (40)K were found in riverbank soil and sediment samples of Netravathi River. In Kali River, the highest (226)Ra activity was recorded for rock samples. To assess the radiological hazard of natural radioactivity in the samples, absorbed gamma dose rates in air, radium equivalent activity, representative level index, external hazard index and internal hazard index associated with the radionuclides were calculated and compared with internationally recommended values. The representative level index (I(gammar)) values are high in sediment samples of Netravathi River. The radium equivalent activity (Ra(eq)), external hazard index (H(ex)) and internal hazard index (H(in)) values are high in rock samples of Kali River. The results of these investigations are presented and discussed in this paper. PMID- 17400346 TI - Kinetics of transfemoral amputees with osseointegrated fixation performing common activities of daily living. AB - BACKGROUND: Direct anchorage of a lower-limb prosthesis to the bone through an implanted fixation (osseointegration) has been suggested as an excellent alternative for amputees experiencing complications from use of a conventional socket-type prosthesis. However, an attempt needs to be made to optimize the mechanical design of the fixation and refine the rehabilitation program. Understanding the load applied on the fixation is a crucial step towards this goal. METHODS: The load applied on the osseointegrated fixation of nine transfemoral amputees was measured using a load transducer, when the amputees performed activities which included straight-line level walking, ascending and descending stairs and a ramp as well as walking around a circle. Force and moment patterns along each gait cycle, magnitudes and time of occurrence of the local extrema of the load, as well as impulses were analysed. FINDINGS: Managing a ramp and stairs, and walking around a circle did not produce a significant increase (P>0.05) in load compared to straight-line level walking. The patterns of the moment about the medio-lateral axis were different among the six activities which may reflect the different strategies used in controlling the prosthetic knee joint. INTERPRETATIONS: This study increases the understanding of biomechanics of bone-anchored osseointegrated prostheses. The loading data provided will be useful in designing the osseointegrated fixation to increase the fatigue life and to refine the rehabilitation protocol. PMID- 17400347 TI - Harmful effects of atmospheric nitrous acid on the physiological status of Scots pine trees. AB - An open top chamber experiment was carried out in the summer of 2003 to examine the effect of nitrous acid (HONO) gas on the physiological status of Scots pine saplings (Pinus sylvestris). Four-year-old pine trees were exposed to two different levels of HONO gas (at ca. 2.5 ppb and 5.0 ppb) and a control (filtered air) from early evening to early morning (18:00-6:00), in duplicate open top chambers. Significant decreases in the ratios of chlorophylls a to b, an increase in the carbon to nitrogen (C/N) ratio, and a reduction of maximum yield of PS II (F(v)/F(m)) in pine needles were also observed after the 2 months' fumigation. Cation contents of pine needles were also decreased by the fumigation with HONO gas. The results could be explained by the harmful effects of OH radicals, generated from photolysis of HONO gas, and/or aqueous phase HONO (NO(2)(-)/HONO), on the photosynthetic capacity of pine needles. PMID- 17400348 TI - Physiological response of the lichen Phaeophyscia hispidula (Ach.) Essl., to the urban environment of Pauri and Srinagar (Garhwal), Himalayas, India. AB - The present study was designed with an aim to observe the effect of increasing urbanization and traffic activity on the physiology of a foliose lichen, Phaeophyscia hispidula (Ach.) Essl., collected from 13 different localities, growing in their natural habitat, in Pauri and Srinagar, two cities in the Himalayas. Six parameters i.e., Chl. a, Chl. b, total pigment, chlorophyll degradation, carotenoid and total protein content, proved the most useful to assess air pollution, were measured. Chlorophyll content and protein content are an efficient parameter to measure the air quality of a region. The study indicates that P. hispidula is pollution tolerant (adaptation) and able to withstand local emissions from vehicle exhausts. PMID- 17400349 TI - Polycyclic aromatic hydrocarbons in leaf cuticles and inner tissues of six species of trees in urban Beijing. AB - Leaf samples of six tree species were collected along urban roadsides and a campus site in Beijing for measurement of polycyclic aromatic hydrocarbons (PAHs). PAHs in leaves were attributed to two fractions, leaf cuticles and inner leaf tissues, using sequential extraction. Total concentrations of 16 PAHs in the cuticles and the inner tissues were 69.3+/-64.6 microg g(-1) (d.w.) and 1.07+/ 0.2 microg g(-1) (d.w.) at roadside and 57.5+/-52.6 microg g(-1) and 0.716+/-0.2 microg g(-1) on campus, respectively. The lipid-normalized inner tissue PAHs varied from 5.8 microg g(-1) to 15.0 microg g(-1). Similarities in PAH spectra between leaf cuticles and airborne particles and between the inner tissues and gaseous phase imply that airborne particulates and gaseous PAHs are likely the sources of PAHs for cuticles and the inner tissues, respectively. Difficulty in migration of heavier PAHs into inner tissues could be another reason. PMID- 17400350 TI - Influence of hydropower dams on the composition of the suspended and riverbank sediments in the Danube. AB - Large hydropower dams have major impacts on flow regime, sediment transport and the characteristics of water and sediment in downstream rivers. The Gabcikovo and Iron Gate dams divide the studied Danube transect (rkm 1895-795) into three parts. In the Gabcikovo Reservoir (length of 40km) only a part of the incoming suspended sediments were deposited. Contrary to this, in the much larger Iron Gate backwater zone and reservoir (length of 310km) all riverine suspended sediments were deposited within the reservoir. Subsequently, suspended sediments were transported by tributaries into the Iron Gate backwater zone. Here they were modified by fractional sedimentation before they transgressed downstream via the dams. Compared with undammed Danube sections, Iron Gate reservoir sediment and suspended matter showed higher clay contents and different K/Ga and Metal/Ga ratios. These findings emphasize the importance of reservoir-river sediment fractionation. PMID- 17400351 TI - Quantifying the importance of diffuse minewater pollution in a historically heavily coal mined catchment. AB - There has been considerable progress in developing treatment systems for point sources of minewater pollution in recent years; however, there remains a knowledge gap in the characterisation and remediation of diffuse minewater sources. Data are presented from the River Gaunless catchment, a historically heavily coal mined catchment in the northeast of England. Instream iron (Fe) loadings were monitored alongside loadings arising from point minewater discharges over a 12-month period to assess the dynamic importance of diffuse sources of minewater pollution. In low flow, diffuse sources account for around 50% of instream loading, a proportion which increases to 98% in high flow conditions. The low flow sources appear to be dominated by direct discharge of contaminated groundwater to surface waters in lower reaches of the catchment. In high flow, resuspended Fe-rich sediments, which are both naturally occurring and derived from historic mining, become the dominant diffuse source of Fe in the water column. PMID- 17400352 TI - Social capital and rates of gonorrhea and syphilis in the United States: spatial regression analyses of state-level associations. AB - We conducted spatial regression analysis to account for spatial clustering of sexually transmitted diseases (STDs) and to examine the state-level association between social capital (using Putnam's public use data set) and rates of gonorrhea and syphilis. We conducted the analysis for the 48 contiguous states of the United States for 1990, 1995, and 2000 and controlled for the effects of regional variation in STD rates, and for state variation in poverty, income inequality, racial composition, and percentage aged 15-34 years. We compared the results of the spatial regression analysis with those of ordinary least squares (OLS) regression. Controlling for all population-level variables, the percentage of variation explained by the OLS regression and by the spatial regression were similar (mid-90s for gonorrhea and low-70s for syphilis), the standardized parameter estimates were similar, and the spatial lag parameter was not statistically significant. Social capital was not associated with STD rates when state variation in racial composition was included in the regression analysis. In this analysis, states with a higher proportion of residents who were African American had higher STD rates. When we did not control for racial composition, regression analysis showed that states with higher social capital had lower STD rates. We conjecture that sexual networks and sexual mixing drive the association between social capital and STD rates and highlight important measurement and research questions that need elucidation to understand fully the relationship between social capital and STDs. PMID- 17400353 TI - Distancing the mad: Jarvis's Law and the spatial distribution of admissions to the Hamilton Lunatic Asylum in Canada, 1876-1902. AB - The institutional confinement of the 'insane' in the nineteenth century constitutes one of the most controversial events in the social history of medicine. Within this scholarship there has emerged an important debate over the spatial determinants of institutionalization. Some studies uphold an historical postulate--Jarvis's Law--that contends there was a 'distance decay' effect in mental hospital utilization--that is, an inverse correlation between the distance from a medical institution and the likelihood of people to use its resources. Other scholars have challenged or modified this thesis, arguing that factors such as the local politics, urban living, or socio-economic status were more important determinants of institutional confinement. This article contributes to this ongoing debate by analysing over 4000 admissions to the Ontario Provincial Asylum in the city of Hamilton, Canada, between 1876 and 1902. The results confirm Jarvis' Law was applicable to the Hamilton context: there was an inverse statistical relationship between physical distance from the asylum and the likelihood of admission. However, this paper yields three additional dimensions to the literature: (1) it demonstrates that jails were much more likely to be utilized as temporary places of confinement for communities far from provincial mental hospitals; (2) the length of stay in the asylum was positively correlated with the distance travelled to the institution; and (3) an inverse relationship was found when correlating distance from the asylum and the likelihood of being readmitted to the same institution. These findings suggest an impact of 'distance' beyond the dimension of hospital utilization and imply that the broader asylum experience could be affected by the previous location of patients. PMID- 17400354 TI - Non-lateralised deficits of drawing production in hemispatial neglect. AB - Spatially lateralised deficits that typically define the hemispatial neglect syndrome have been shown to co-occur with other non-lateralised deficits of attention, memory, and drawing. However even a simple graphic task involves multiple planning components, including the specification of drawing start position and drawing direction. In order to investigate the influence of these factors in neglect we presented patients with a circle-copying task, and specified the drawing start point. The ability to draw from the instructed location was strongly related to tests that measure constructional abilities, but not related to start point laterality. In contrast, the direction in which patients drew the circle was affected by start point laterality: patients with neglect were less likely to draw in a typical direction when the cue was on the affected side of space and this was strongly related to severity of the neglect. Patients with neglect consistently produced circles that were smaller than the model; however, the scaling was not affected by the laterality of the start point, nor was the proportion of drawings correctly started at the cue. These findings demonstrate the complex relationship between neglect and even the simplest test for the syndrome. PMID- 17400355 TI - Signs and symptoms related to temporomandibular disorders--Follow-up of subjects with shortened and complete dental arches. AB - OBJECTIVE: To assess prevalence of cardinal signs and symptoms related to temporomandibular disorders (TMD) in subjects with shortened dental arches and to clarify the individual course of these signs and symptoms. METHODS: In this 9 year follow-up study, subjects with shortened dental arches (n=74) were compared with subjects with complete dental arches (n=72). Of three reported symptoms (pain, noises/clicking and restricted mobility of the lower jaw), and of two clinical signs (palpated clicking of the temporomandibular joint and restricted maximal mouth opening) estimates of mean scores were calculated by a mixed model. Suggested determinants for TMD (gender, time, bruxism, chewing side preference) were investigated by covariate analyses. Subjects with complete 9-year follow-up (shortened dental arches: n=42; complete dental arches: n=41) were described more detailed by frequency distributions, Pearson correlations of signs and symptoms, and fluctuation of the symptoms. RESULTS: Covariate analyses using the mixed model revealed no significant differences between the shortened and the complete dental arch groups (p>0.05) with respect to symptoms and signs. Most prevailing effect was gender: females reported more frequently pain (p=0.05) and noises/clicking (p=0.03). Restricted mobility was significantly related with chewing side preference and bruxism habits (both: p=0.01). In both groups, subjects with complete 9-year follow-up had low prevalence of serious symptoms and signs and symptoms fluctuated without demonstrable correlation. CONCLUSION: In this 9-year follow-up, subjects with shortened dental arches had similar prevalence, severity, and fluctuation of signs and symptoms related to TMD compared to subjects with complete dental arches. PMID- 17400357 TI - Oligosaccharide structure of a functional unit RvH1-b of Rapana venosa hemocyanin using HPLC/electrospray ionization mass spectrometry. AB - In the present study the structures of two glycopeptides (G1 and G1'), isolated from FU RvH(1)-b and two glycopeptides (G2 and G3), isolated from the structural subunit RvH(1) of Rapana venosa hemocyanin, were determined. To structurally characterize the site-specific carbohydrate heterogeneity and binding site of the N-linked glycopeptide(s), a combination of capillary reversed-phase chromatography and ion trap mass spectrometry was used. The amino acid sequences of glycopeptides G1 and G1' determined by Edman degradation and MS/MS sequencing demonstrated that the oligosaccharides are linked to N-glycosylation sites. Two peptides (a glycosylated (G1) and non-glycosylated one) were identified in this fraction and no linkage sites were observed in the latter one. Based on the sequencing of the glycosylated fractions G1, G1', G2 and G3, the carbohydrate structure Man(alpha1-6)Man(alpha1-3)Man(beta1-4)GlcNAc(beta1-4)[Fuc(alpha1 6)]GlcNAc-R could be identified for glycopeptides G1 and G3, and only the typical core structure Man(alpha1-6)Man(alpha1-3)Man(beta1-4)GlcNAc(beta1-4)GlcNAc-R was found for G1' and G2. The Fuc residue found in glycopeptides G1 and G3 is attached to N-acetyl-glucosamine of the carbohydrate core, as often found in other glycoproteins. PMID- 17400356 TI - Diverse functions of RNase L and implications in pathology. AB - The endoribonuclease L (RNase L) is the effector of the 2-5A system, a major enzymatic pathway involved in the molecular mechanism of interferons (IFNs). RNase L is a very unusual nuclease with a complex mechanism of regulation. It is a latent enzyme, expressed in nearly every mammalian cell type. Its activation requires its binding to a small oligonucleotide, 2-5A. 2-5A is a series of unique 5'-triphosphorylated oligoadenylates with 2'-5' phosphodiester bonds. By regulating viral and cellular RNA expression, RNase L plays an important role in the antiviral and antiproliferative activities of IFN and contributes to innate immunity and cell metabolism. The 2-5A/RNase L pathway is implicated in mediating apoptosis in response to viral infections and to several types of external stimuli. Several recent studies have suggested that RNase L could have a role in cancer biology and evidence of a tumor suppressor function of RNase L has emerged from studies on the genetics of hereditary prostate cancer. PMID- 17400358 TI - NOX family NADPH oxidases: not just in mammals. AB - NOX family NADPH oxidases are enzymes whose biological function is electron transport and the generation of reactive oxygen species (ROS). NOX enzymes in mammalian organisms have received most attention. However, NOX enzymes are widely distributed in different kingdoms of life. While they are not found in prokaryotes and most unicellular eukaryotes, they are present in fungi, plants, and animals. The identity of the ancestral NOX is not known, but most likely it: (i) possessed the basic NOX structure consisting of 6 transmembrane domains (containing two assymmetrical hemes) and a long cytoplasmic C-terminal (containing the FAD and NADPH binding sites); and (ii) emerged before the divergence of life into fungi, plants, and animals. During evolution, acquisition of a Ca(2+)-binding EF hand domain by an ancestral NOX, led to NOX5-like isoforms. DUOX isoforms presumably developed from a NOX5-like isoform through the additional acquisition of a peroxidase homology domain. The expression pattern of NOX enzymes is specific to each kingdom of life. Fungi express only ancestral type isoforms, and plants only NOX5-like isoforms. NOX expression patterns in animals are complex and ancestral NOXes, NOX5-like isoforms and DUOXes are generally found. But there are exceptions; for example rodents lack NOX5 and Caenorhabditis elegans expresses only DUOXes. Biological functions of NOX enzymes include, among others, host defense, post-translational modification of proteins, and regulation cell growth and differentiation. In summary, the invention of NOX enzymes early in the development of life was a success story: there is no evidence of multicellular life without NOX enzymes. PMID- 17400359 TI - Adipogenesis-related increase of semicarbazide-sensitive amine oxidase and monoamine oxidase in human adipocytes. AB - A strong induction of semicarbazide-sensitive amine oxidase (SSAO) has previously been reported during murine preadipocyte lineage differentiation but it remains unknown whether this emergence also occurs during adipogenesis in man. Our aim was to compare SSAO and monoamine oxidase (MAO) expression during in vitro differentiation of human preadipocytes and in adipose and stroma-vascular fractions of human fat depots. A human preadipocyte cell strain from a patient with Simpson-Golabi-Behmel syndrome was first used to follow amine oxidase expression during in vitro differentiation. Then, human preadipocytes isolated from subcutaneous adipose tissues were cultured under conditions promoting ex vivo adipose differentiation and tested for MAO and SSAO expression. Lastly, human adipose tissue was separated into mature adipocyte and stroma-vascular fractions for analyses of MAO and SSAO at mRNA, protein and activity levels. Both SSAO and MAO were increased from undifferentiated preadipocytes to lipid-laden cells in all the models: 3T3-F442A and 3T3-L1 murine lineages, human SGBS cell strain or human preadipocytes in primary culture. In human subcutaneous adipose tissue, the adipocyte-enriched fraction exhibited seven-fold higher amine oxidase activity and contained three- to seven-fold higher levels of mRNAs encoded by MAO A, MAO-B, AOC3 and AOC2 genes than the stroma-vascular fraction. MAO-A and AOC3 genes accounted for the majority of their respective MAO and SSAO activities in human adipose tissue. Most of the SSAO and MAO found in adipose tissue originated from mature adipocytes. Although the mechanism and role of adipogenesis-related increase in amine oxidase expression remain to be established, the resulting elevated levels of amine oxidase activities found in human adipocytes may be of potential interest for therapeutic intervention in obesity. PMID- 17400360 TI - Modeling military trampling effects on glacial soils in the humid continental climate of southern New York. AB - The purpose of this research is to create a baseline model of soil compaction response to trampling and a methodology to model the effects of trampling on soil. Although trampling studies have been conducted in the past, the analysis of military training in part provides a different perspective and approach. The data showed bulk densities remained relatively constant for a time and then began to increase at an increasing rate for several hundred passes and finally leveled and remained at or below 1.30 g/cm3 through the remainder of the experiment. Mathematical models were created based on empirical data from a trampling experiment using a more standard logistical growth curve as well as curves based on Weibull and gamma cumulative distribution functions (CDFs). The experiment and the resulting models give quantifiable continuous inference on the effects of trampling, as opposed to the existing qualitative assessments. These baseline models will be the foundation for future studies of land management when trampling occurs. PMID- 17400361 TI - Re: Masashi Yano, Takashi Imamoto, Hiroyoshi Suzuki et Al. The clinical potential of pretreatment serum testosterone level to improve the efficiency of prostate cancer screening. Eur urol 2007;51:375-80. PMID- 17400362 TI - Re: Matthew E. Nielsen, Shahrokh F. Shariat, Pierre I. Karakiewicz et al. Advanced age is associated with poorer bladder cancer-specific survival in patients treated with radical cystectomy. Eur urol 2007;51:699-708. PMID- 17400364 TI - Combined percutaneous radiofrequency ablation and ethanol injection of renal tumours: midterm results. AB - OBJECTIVES: To evaluate the safety and efficacy of combined percutaneous, image guided, radiofrequency (RF) ablation and ethanol injection of renal tumours, and to present our midterm results. METHODS: Since February 2002, 27 consecutive patients (22 men, 5 women; age range: 39-84 yr; mean: 69) with 28 renal tumours (mean diameter: 2.87cm) were treated with combined percutaneous RF and ethanol ablation, and were prospectively evaluated. Twenty-five patients were considered nonsurgical candidates because of comorbid conditions (16 patients) or had previous nephrectomy (9 patients), and 2 had refused surgery. Thirty-three ablation sessions were performed, with computed tomography (26 sessions), ultrasound (6), or combined magnetic resonance imaging/fluoroscopic guidance in 1. Absolute ethanol (0.5-3ml; mean: 1.7) was injected into the tumour immediately before treatment with radiofrequency. Mean follow-up period was 18.6 mo (range: 3 56). RESULTS: Twenty-seven of the 28 tumours were completely ablated with either one (21 tumours) or two treatment sessions (6 tumours). One patient with residual disease refused further treatment. Only three minor complications, including a subcapsular haematoma and two patients with loin pain, occurred; all three patients were treated conservatively. None of the complications was related to the ethanol injection. During the follow-up period, no evidence of local recurrence or metastatic disease was seen. Creatinine levels have not changed significantly in any of the patients following ablation. CONCLUSIONS: Combined use of percutaneous RF and ethanol ablation is a safe and effective alternative treatment for selective patients with renal tumours. PMID- 17400365 TI - Robotic-assisted laparoscopic radical cystoprostatectomy. AB - OBJECTIVES: Recent small case series have been reported for robotic-assisted laparoscopic radical cystoprostatectomy. The present literature includes 34 patients who have undergone robotic-assisted cystectomy procedures. We report our initial experience with robotic-assisted laparoscopic radical cystoprostatectomy, describing stepwise the surgical procedure and evaluating perioperative and pathologic outcomes of this novel procedure. METHODS: Twenty men underwent robotic-assisted laparoscopic radical cystoprostatectomy and extracorporeal urinary diversion for clinically localized bladder cancer. The stepwise operative procedure is described in detail. Outcome measures evaluated included operative variables, hospital recovery, pathologic outcomes, and complication rate. Comparisons were made to these gender-matched 24 men who underwent an open procedure during this same period. RESULTS: Mean age was 62.3 yr (range: 54-76 yr). Ten patients underwent ileal conduit diversion and 10 patients underwent an orthotopic neobladder. In all cases the urinary diversion was performed extracorporeally. Mean operating room time of all patients was 6.1h (most recent 10 cases, 5.2h). Mean surgical blood loss was 313 ml. On surgical pathology, 14 patients were < or =pT2, 4 patients pT3, and 2 patients N+. In no case was there inadvertent entry into the bladder or positive surgical margins. Mean number of lymph nodes removed was 19 (range: 6-29). Mean time to flatus was 2.1 d and bowel movement 2.8 d. Sixteen patients were discharged on postoperative day (POD) 4, three patients on POD 5, and one on POD 8. There were six postoperative complications (30%) in five patients. CONCLUSIONS: Our initial experience with robotic-assisted laparoscopic radical cystoprostatectomy appears to be favorable with acceptable operative, pathologic, and short-term clinical outcomes. As our experience increases, we should expect to continue to refine our surgical technique and reduce operating room times. Larger experiences are required to adequately evaluate and validate this procedure as an appropriate surgical and oncologic option for the bladder cancer patient. PMID- 17400366 TI - Extracorporeal shock wave lithotripsy of lower calyx calculi: how much is treatment outcome influenced by the anatomy of the collecting system? AB - OBJECTIVES: Extracorporeal shock wave lithotripsy (ESWL) of lower calyx stones has been criticized because of the high incidence of residual fragments. Controversial results have been reported regarding the stone-free rate after ESWL depending on the influence of the collecting system anatomy on stone clearance. Therefore we evaluated our stone-free rate after ESWL of lower calyx stones and searched for correlations to various anatomic parameters of the collecting system. METHODS: Ninety-six patients with isolated lower calyx stones treated exclusively with ESWL (Dornier HM3) were evaluated 3 mo postoperatively. The results were correlated with the following anatomic parameters of the collecting system as determined from the pretreatment intravenous urography: (1) lower infundibulum width, (2) lower infundibulum length, (3) infundibulopelvic angle, (4) volume of the collecting system. Follow-ups were performed 24 h after ESWL with an abdominal plain film and 3 mo postoperatively with a urography or abdominal plain X-ray together with renal ultrasound. RESULTS: Three months postoperatively, 68% of all patients were stone free, including 69% of the patients with stones initially < or =1 cm, and 67% of the patients with stones >1 cm. Stone-free patients compared with patients having residual fragments had no significant differences in infundibulum width, infundibulum length, infundibulopelvic angle, or collecting system volume. CONCLUSIONS: A stone-free rate 3 mo after ESWL of 68% overall justifies ESWL as a possible treatment option for lower calyx stones. Influence of the collecting system anatomy on disintegrate clearance from the lower calyx could not be demonstrated. PMID- 17400367 TI - Toward an adequate mathematical model of mental space: conscious/unconscious dynamics on m-adic trees. AB - We try to perform geometrization of cognitive science and psychology by representing information states of cognitive systems by points of mental space given by a hierarchic m-adic tree. Associations are represented by balls and ideas by collections of balls. We consider dynamics of ideas based on lifting of dynamics of mental points. We apply our dynamical model for modeling of flows of unconscious and conscious information in the human brain. In a series of models, Models 1-3, we consider cognitive systems with increasing complexity of psychological behavior determined by structure of flows of associations and ideas. PMID- 17400368 TI - Myoclonic epilepsy masquerading as a tic disorder. AB - Myoclonic epilepsies comprise a heterogeneous group of epileptic syndromes characterized by myoclonic jerks, generalized tonic-clonic seizures (GTCSs), and sometimes absence seizures (multiplicity of seizure types). JME (Juvenile Myoclonic Epilepsy) is relatively common and usually responds well to treatment with appropriate anticonvulsants. Others like the progressive myoclonic epilepsy syndromes are relentlessly progressive, associated with cognitive decline and respond poorly to anticonvulsants. Myoclonic epilepsy is frequently misdiagnosed until the patient is specifically asked about the leading symptom, i.e., jerky movements occurring in the morning. We present here a patient who had been misdiagnosed as a tic disorder and treated for the past many years before the correct diagnosis of myoclonic epilepsy was made during an admission for Video EEG monitoring. PMID- 17400369 TI - Postoperative relieve of abnormal vasoregulation in carpal tunnel syndrome. AB - OBJECTIVE: To study the skin temperature changes as a sign of altered blood flow regulation due to abnormal sympathetic nerve function in carpal tunnel syndrome (CTS) and the efficacy of carpal tunnel release (CTR) operation in the alleviation of those signs and other symptoms of CTS. SUBJECTS AND METHODS: Forty one healthy hands (n=41) of 22 volunteers and 22 hands (n=22) with clinically diagnosed carpal tunnel syndrome of 16 patients were examined. A series of infrared photos of the hands of each subject were taken and stored by using digital infrared thermography (DIRT) before and 6 months after the CTR. The temperatures of the finger tips from digit 1 (D1) to digit 5 (D5), the center point of thenar (Th) and hypothenar (Ht) eminences were measured, the median nerve index (MI=(D1-D2)+(D1-D3)+(D2-D3)) and the temperature differences between the median and the ulnar nerve distribution area (MED.ULN=(D1-D5)+(D2-D5)+(D3 D5)+(Th-Ht)) were calculated. Absolute values were used when calculating the temperature differences between different points. RESULTS: All the CTS patients were released from the CTS symptoms, i.e. numbness and pain after the operation, and the results of nerve conduction studies (NCS) were all back to normal. The MI and MED.ULN in CTS hands were significantly smaller before CTR compared to those of healthy control subjects (MI: p<0.001, MED.ULN: p<0.005), and those after CTR (p<0.005 for both parameters). The results also showed that all finger tips of the affected hand including those innervated by the ulnar nerve were cooler before but warmer after the operation. Before the operation, the non-CTS hands of the CTS patients, were also colder (although the difference was not significant) than the healthy control subjects but approached the same level after the CTR operation of the CTS hands. CONCLUSION: The results of DIRT measurements suggest that the blood flow regulation in CTS is abnormal possibly because of disturbed sympathetic vasomotor regulation and that the circulation gets normal together with the alleviation of the other symptoms of CTS as recorded 6 months after CTR operation. PMID- 17400370 TI - Experimental hypoxia is a potent stimulus for radiotracer uptake in vitro: comparison of different tumor cells and primary endothelial cells. AB - Hypoxia causes upregulation of vascular endothelial growth factor (VEGF) which is a key regulator in tumor angiogenesis and essential for the proliferation of endothelial cells. Endothelial cells have been described to accumulate radiotracers like (18)F-FDG. However, the contribution of radiotracer uptake by endothelial cells to uptake measured in tumors by positron emission tomography (PET) is still unclear. In this study (18)F-FDG and (18)F-FMISO radiotracer uptake in various tumor and primary endothelial cells cultured at hypoxic conditions was investigated. Experimental hypoxia was confirmed by significant upregulation of VEGF mRNA. In comparison to normoxic conditions, cellular uptake of (18)F-FDG was significantly increased at hypoxic conditions in two of the tumor and all endothelial cells, whereas (18)F-FMISO uptake was only enhanced in tumor cell lines HT-29 and MCF-7. Our data showed a marked influence of experimental hypoxia on the metabolism and gene expression of tumor and endothelial cells in vitro. This indicates an important contribution of endothelial cells to (18)F-FDG radiotracer uptake in tumors and for the visualization of tumors by means of PET. PMID- 17400371 TI - Biosorption of Astrazone Blue basic dye from an aqueous solution using dried biomass of Baker's yeast. AB - In this study dried biomass of Baker's yeast, Saccharomyces cerevisiae, is used as a sorbent for Astrazone Blue basic dye aqueous solution. Factors affecting the adsorption process: dye concentration, contact time, temperature and pH were investigated. The equilibrium concentration and the adsorption capacity at equilibrium were determined using three different sorption models namely: Langmuir, Freundlich and Temkin isotherms. It was found that increasing temperature and pH result in higher dye loadings per unit weight of the sorbent. The results gained from this study were described by Langmuir isotherm model better than Freundlich and Temkin isotherm models. The calculated heat of adsorption of the dye-yeast system indicates that the bio-sorption process is taking place by chemical adsorption and has an endothermic nature. The maximum adsorption capacity at 30 degrees C and pH 7 was calculated as 70 mg/g for dried biomass of Baker's yeast compared to 18.5mg/g for commercial granular activated carbon, indicating that dried biomass of Baker's yeast can be considered as a good sorbent material for Astrazone Blue solution. PMID- 17400372 TI - Sorption of aqueous phosphorus onto bituminous and lignitous coal ashes. AB - Aiming at the development of a phosphorus removal technology for waste water, phosphate (PO(4)(3-)) retention behavior of bituminous and lignitous coal ashes was investigated using a batch reactor. Ash samples, including fresh and weathered fly and bottom ashes, were studied for their sorption isotherms and reversibility. Fly ashes had a much higher phosphate retention capacity (4000 30,000mgP/kg) than bottom ashes (15-600mgP/kg). Lignitous coal ashes were more capable of retaining phosphate than bituminous coal ashes. The retention process was largely irreversible, and the irreversibility increased with the increase in the retention capacity. Weathering enlarged the retention capacity of the bituminous bottom ash, but substantially lowered that of the fly ash, likely due to the difference in the weather-induced changes between the fly and bottom ashes. Sorption isotherms of fly ashes were found to be adequately represented by the Langmuir model while those of bottom ashes fitted better to the Freundlich model. Concentrations of Ca(2+) and PO(4)(3-) in the aqueous phase were measured at the end of sorption and desorption experiments, and were compared with solubilities of three calcium phosphate minerals. The aqueous solutions were saturated or super-saturated with respect to tricalcium phosphate (Ca(3)(PO(4))(2)) and hydroxyapatite (Ca(5)(PO(4))(3)OH), and slightly under saturated with respect to amorphous calcium phosphate. It is concluded that precipitation of calcium phosphate is the predominant mechanism for phosphate retention by coal ash under the conditions studied. There is a strong and positive correlation between alkalinity and phosphate sorption capacity. Consequently, acid neutralization capacity (ANC) can be used as an indicator of phosphate sorption capacity of coal ashes. PMID- 17400373 TI - Toxic elements at a disused mine district: Particle size distribution and total concentration in stream sediments and mine tailings. AB - Heavy metal and metalloid pollution at a disused pyrite mine was investigated. Five solid samples collected in the area (three stream sediments with different soil texture, a background soil sample and a mine tailing) were characterised by mineral and element composition, particle size distribution (by wet and dry sieving and laser diffraction) and total concentration by acid digestion (Mn, Zn, Cu, Pb, Hg, Cd, Sb and As). X-ray and element analyses denoted a common mineralogical and chemical composition of mainly quartz, clinochlore, muscovite, anorthite, and hematite. Particle size distributions of the five samples showed that stream sediments were characterised by larger percentage of sand range classes (2000-60 microm) while background sample and tailing are mainly characterised by gravel particles (>2000 microm). Wet and dry sieving procedures gave different particle size distributions, which can be interpreted by laser diffraction analysis and represented by Rosin-Rammler model. Concentrations of Zn, Cu and Cd were higher in the stream sediments than the tailing and background soil, while Mn, As, Sb and Hg are mainly concentrated in the tailing sample. Metal concentrations in the three stream sediment samples are correlated with both particle size dimensions (D(63.2)) and concentration of geochemical normalizers (iron and aluminium). These correlations are observed also for the pollutants that are mainly concentrated in tailing sample (Mn and As), denoting the importance of surface interactions also for the binding of these elements onto stream sediments. PMID- 17400374 TI - Comparative study of UV/TiO2, UV/ZnO and photo-Fenton processes for the organic reactive dye degradation in aqueous solution. AB - In this study advanced oxidation processes (AOPs), UV/TiO(2), UV/ZnO and photo Fenton, were applied in order to degrade C.I. Reactive Red 45 (RR45) dye in aqueous solution. The effects of key operating parameters, such as initial pH, catalyst and hydrogen peroxide dosage as well as the effect of initial dye concentration on decolorization and mineralization extents were studied. Primary objective was to determine the optimal conditions for each of the processes. The influence of added zeolite on the process efficiency was also studied. UV/vis spectrophotometric and total organic carbon (TOC) measurements were performed for determination of decolorization and mineralization extents. It has been found that photo-Fenton process was the most efficient with 74.2% TOC removal and complete color removal achieved after a 1h treatment. PMID- 17400375 TI - Enhanced photocatalytic degradation of pollutants in petroleum refinery wastewater under mild conditions. AB - A circulating photocatalytic reactor was used for removing aliphatic and aromatic organic pollutants in refinery wastewater. The TiO(2) added wastewater samples, while saturating with air, were irradiated with an immersed mercury UV lamp (400 W, 200-550 nm). Optimal catalyst concentration, fluid pH and temperature were obtained at amounts of near 100 mgL(-1), 3 and 318K, respectively. A maximum reduction in chemical oxygen demand of more than 90% was achieved after about 4h irradiation and hence, 73% after about only 90 min; significant pollutant removal was also achievable in the other conditions. The identification of the organic pollutants, provided by means of a GC/MS and a GC analysis systems, equipped with headspace injection technique, showed that the major compounds were different fractions of petroleum aliphatic hydrocarbons (up to C(10)) and the well-known aromatic compounds such as benzene, toluene and ethylbenzene. The results showed a high efficiency degradation of all of these pollutants. PMID- 17400376 TI - Self-triggering reaction kinetics between nitrates and aluminium powder. AB - During the night between the 19 and 20 September 2003, a loud explosion occurred at about 3km from the town of Carignano that was clearly heard at a distance of some tens of kilometres. The explosion almost completely destroyed most of the laboratories of the Panzera Company that were used for the production of fireworks. The results of the research activities that were carried out using a differential scanning calorimeter (DSC) on the same raw materials that made up the pyrotechnical mixture that exploded are reported in this paper. This activity was carried out to identify the dynamics of the accident. It proved possible to verify how the event was produced because of a slow exothermic reaction which, after about 8h, caused the self-triggering of 120 kg of finished product. The detonation can therefore be put down to a runaway reaction in the solid phase, whose primogenial causes can be attributed to a still craftsman type production system, not conformed to the rigorous controls and inspections as those required by a safety management system for major risk plants, as the Panzera Company was. PMID- 17400377 TI - Glutathione depletion induces differential apoptosis in cells of mouse retina, in vivo. AB - Oxidative stress affects numerous intracellular macromolecules, and may result in cell death unless precisely regulated. Unregulated oxidative stress can be controlled by various cellular defense mechanisms such as glutathione (GSH) which can critically counteract the damaging effects of oxidative stress in mammalian cells. We determined the effects of unregulated oxidative stress induced by GSH depletion on cells in mouse retina. Mice were intraperitoneally injected with buthionine sulphoximine (BSO) at 1.5 g/kg. After 0, 1, 4, and 7 days of BSO administration, retinas were excised and sections were subjected to GSH assay and terminal uridine deoxynucleotidyl nick end labeling (TUNEL) analysis. After 4 days of BSO administration, the number of TUNEL positive cells was significantly increased. However, after 7 days, TUNEL positive cells returned to the basal level. The retinal region most affected by the BSO treatment appeared to be the outer nuclear layer where the photoreceptor cells reside. Different from cells in other regions, retinal cells in the inner nuclear layer increased in their apoptosis even after the first day of BSO injection, and the increase was further potentiated after 4 days. Taken together, our studies suggested that GSH depletion may cause unregulated oxidative stress to the cells in the retina and indeed increased cell death in the retina. The cells in the inner nuclear layer seemed to be affected earlier than the cells in other layers of the retina. The GSH level in the retina may be a crucial therapeutic target in preventing blindness. PMID- 17400378 TI - Wnt expression in the adult rat subventricular zone after stroke. AB - Neurogenesis occurs in adult brain neural progenitor cells (NPCs) located in the subventricular zone (SVZ) of the lateral ventricle and the subgrandular zone of the hippocampal dentate gyrus. After ischemic stroke, NPCs in the SVZ proliferate and migrate towards the ischemic boundary region to replenish damaged neurons. During development, the Wnt pathways contribute to stem cell maintenance and promote neurogenesis. We hypothesized that stroke up regulates Wnt family genes in SVZ cells. Non-ischemic and ischemic cultured SVZ cells and a single population of non-ischemic and ischemic SVZ cells isolated by laser capture microdissection (LCM) were analyzed for Wnt pathway expression using real-time RT PCR and immunostaining. The number of neurospheres increased significantly (p<0.05) in SVZ cells derived from ischemic (32+/-4.7/rat) compared with the number in non-ischemic SVZ cells (18+/-3/rat). Wnt family gene mRNA levels were detected in SVZ cells isolated from both cultured and LCM SVZ cells, however there was no up regulation between non-ischemic and ischemic SVZ cells. Immunostaining on brain sections also demonstrated no up regulation of Wnt pathway protein between ischemic and non-ischemic SVZ cells. Expression of the Wnt family genes in SVZ cells support the hypothesis that the Wnt pathway may be involved in neurogenesis in the adult brain. However, ischemia does not up regulate Wnt family gene expression. PMID- 17400379 TI - Neuroprotective effects of phenylenediamine derivatives independent of an antioxidant pathway in neuronal HT22 cells. AB - Resistance to oxidative stress often determines neuronal survival in the brain. Thus, antioxidants are supposed to be promising neuroprotective compounds against neurodegenerative diseases. For example, N,N'-diphenyl-p-phenylenediamine (DPPD) reportedly exerts cytoprotective effects against oxidative stress possibly by acting as an antioxidant. DPPD can give electron(s) to free radicals and thus scavenge them, and protect the cells from oxidative stress. The antioxidative activities of DPPD are prominent at the micromolar order, but what about its effects at much lower concentrations? We concluded that DPPD has two actions on neuronal cells, antioxidant activity and an unknown neuroprotective effect, which are effective at micromolar and nanomolar levels, respectively. In the present report, we found that DPPD inhibited cell death caused by oxidative stress at nanomolar order (1/1000 lower than concentrations needed for antioxidant activity) and that the effects were independent of antioxidant activities. DPPD inhibited the oxidative glutamate toxicity but not the tumor necrosis factor alpha-, hydrogen peroxide-, or xanthine+xanthine oxidase-induced death of HT22 cells, a mouse neuronal cell line. DPPD and phenylenediamine derivatives protected HT22 cells against oxidative glutamate toxicity at nanomolar concentrations. By studying the structure-function relationship of these compounds, we found the structure of phenyl-amine-phenyl-amine-phenyl (or butyl) to be essential for the neuroprotective effects. PMID- 17400380 TI - Tau in cerebrospinal fluid: a sensitive sandwich enzyme-linked immunosorbent assay using tyramide signal amplification. AB - Tau in cerebrospinal fluid (CSF) has been proposed as a diagnostic marker for Alzheimer's disease (AD). This paper presents a new sensitive sandwich ELISA allowing quantitation of tau from 8 microl CSF/well. A human specific monoclonal tau antibody HT7 was used as a capture antibody and a mixture of polyclonal tau antibodies, 92e and R134d was used as reporter antibodies. Tyramide signal amplification (TSA) technology was used in the last step to increase the sensitivity. With this TSA-ELISA, the lowest detection limit for tau was 14.3 pg/ml. Tau levels in CSF were found to be increased in AD patients (807+/-304 pg/ml, p<0.001) compared with controls (252+/-94 pg/ml). Thirty-five of 38 AD cases (92% sensitivity) yielded signals greater than cutoff, while only 1 of 38 control cases (97% specificity) was greater. A highly significant correlation was found between this assay and a commonly used kit, INNOTEST hTAU Antigen. PMID- 17400381 TI - Spatial frequency characteristics of nearby neurons in cats' visual cortex. AB - Various methods have allowed mapping of responses to several stimulus features on the cortical surface, particularly edge orientation and motion direction. The cortical mapping of spatial frequencies (SF), which is the basic property that leads to perception of spatial details of visual objects, is still controversial. We recorded simultaneously extracellular action potentials from neighboring cells in superficial layers of the area 17-18 border region of anesthetized cats. Responses of nearby cells to sine-wave gratings of varying SF were analyzed. Spatial frequency tuning curves were cross-correlated to establish the degree of similarity between the curves and optimal SFs were compared for each pair of neurons. The investigation showed that only about a half of nearby neurons exhibited close optimal SFs and similar tuning curves. The results suggest that SF channels do not show a clear clustering within a small pool of neurons. Such organization may contribute to the perception of spatial details at all orientations and motion directions. PMID- 17400382 TI - Staurosporine-induced apoptosis in astrocytes is prevented by A1 adenosine receptor activation. AB - Astrocyte apoptosis occurs in acute and chronic pathological processes at the central nervous system and the prevention of astrocyte death may represent an efficacious intervention in protecting neurons against degeneration. Our research shows that rat astrocyte exposure to 100 nM staurosporine for 3h caused apoptotic death accompanied by caspase-3, p38 mitogen-ed protein kinase (MAPK) and glycogen synthase kinase-3beta (GSK3beta) activation. N(6)-chlorocyclopentyladenosine (CCPA, 2.5-75 nM), a selective agonist of A(1) adenosine receptors, added to the cultures 1h prior to staurosporine, induced a dose-dependent anti-apoptotic effect, which was inhibited by the A(1) receptor antagonist, 8-cyclopentyl-1,3 dipropylxanthine. CCPA also caused a dose- and time-dependent phosphorylation/activation of Akt, a downstream effector of cell survival promoting phosphatidylinositol 3-kinase (PI3K) pathway, which in turn led to inhibition of staurosporine-induced GSK3beta and p38 MAPK activity. Accordingly, the anti-apoptotic effect of CCPA was abolished by culture pre-treatment with LY294002, a selective PI3K inhibitor, pointing out the prevailing role played by PI3K pathway in the protective effect exerted by A(1) receptor activation. Since an abnormal p38 and GSK3beta activity is implicated in acute (stroke) and chronic (Alzheimer's disease) neurodegenerative diseases, the results of the present study provide a hint to better understand adenosine relevance in these disorders. PMID- 17400383 TI - Semantic, perceptual and number space: relations between category width and spatial processing. AB - Coarse semantic encoding and broad categorization behavior are the hallmarks of the right cerebral hemisphere's contribution to language processing. We correlated 40 healthy subjects' breadth of categorization as assessed with Pettigrew's category width scale with lateral asymmetries in perceptual and representational space. Specifically, we hypothesized broader category width to be associated with larger leftward spatial biases. For the 20 men, but not the 20 women, this hypothesis was confirmed both in a lateralized tachistoscopic task with chimeric faces and a random digit generation task; the higher a male participant's score on category width, the more pronounced were his left-visual field bias in the judgement of chimeric faces and his small-number preference in digit generation ("small" is to the left of "large" in number space). Subjects' category width was unrelated to lateral displacements in a blindfolded tactile motor rod centering task. These findings indicate that visual-spatial functions of the right hemisphere should not be considered independent of the same hemisphere's contribution to language. Linguistic and spatial cognition may be more tightly interwoven than is currently assumed. PMID- 17400384 TI - Amphetamine reward in the monogamous prairie vole. AB - Recent studies have shown that the neural regulation of pair bonding in the monogamous prairie vole (Microtus ochrogaster) is similar to that of drug seeking in more traditional laboratory rodents. Therefore, strong interactions between social behavior and drug reward can be expected. Here, we established the prairie vole as a model for drug studies by demonstrating robust amphetamine-induced conditioned place preferences in this species. For both males and females, the effects of amphetamine were dose-dependent, with females being more sensitive to drug treatment. This study represents the first evidence of drug reward in this species. Future studies will examine the effects of social behavior on drug reward and the underlying neurobiology of such interactions. PMID- 17400385 TI - Electrophysiological findings of acute peripheral facial palsy in diabetic and non-diabetic patients. AB - The aim of this study is to investigate the role of diabetes mellitus on the clinical and electrophysiological findings of peripheral facial palsy (PFP), the effect of the diabetes duration and polyneuropathy on the electrophysiological parameters. A total of 32 diabetic and 40 non-diabetic patients with peripheral facial palsy were included. All patients were divided into two subgroups based on the time of electrophysiological examinations: within the first 15 days versus within 16-30 days. Neuropathy symptoms and the results of neurological examinations and electrophysiological findings were recorded. The findings of electroneurography (EnoG), blink reflex (BR) evaluation, and needle electromyography (EMG) indicated statistically significant blink reflex abnormalities in diabetic patients compared to non-diabetics. Delay in the latency was more remarkable in the R2 component than in the R1 (p<0.001). The delay in the R1 latency was also observed in the non-affected side for diabetic patients. The longer duration of the diabetes caused significant delay on the blink reflex latency on both the affected and non-affected sides for R1 component (p=0.019, p=0.041, respectively). In contrary, neither the diabetes duration nor the age of the patients correlated with the clinical severity of facial palsy, fiber loss, fibular nerve compound muscle action potential amplitudes, and the nerve conduction velocities. PMID- 17400386 TI - Gene expression changes in the rodent hippocampus following whole brain irradiation. AB - Therapeutic cranial irradiation may result in debilitating cognitive impairments. In human patients these deficits are age and radiation dose-dependent and are attributed to a diminished capability to learn and memorize new tasks and information. Because of the known involvement of the hippocampus in memory consolidation, it is important to identify irradiation-induced changes including alterations in gene expression in this structure. Whole brain irradiation doses of 0, 0.3, 3, 10, or 30 Gray (Gy) were administered to 3-month-old rats in a single session. Twenty-four hours following cranial irradiation, hippocampi were processed for oligonucleotide microarrays analysis. Metallothioneins (MT)-I and II, heat shock protein (Hsp-27), glial fibrillary acidic protein alpha (GFAP), and c-Fos genes were altered significantly across the various doses of irradiation. A pathway analysis shows that these genes were centered around the immediate early gene myc and tumor suppressor gene (TP53). Our results identified important genes and possible pathways that are altered in the hippocampus in the acute phase following cranial irradiation, and implicate gene pathways important for both learning and memory and apoptosis. PMID- 17400387 TI - Coping with fibromialgia: usefulness of the Chronic Pain Coping Inventory-42. AB - There are few studies on coping with fibromyalgia (FM). The aim of the present study was to assess the usefulness of a Spanish version of the Chronic Pain Coping Inventory-42 (CPCI-42) in patients with FM. A random sample (N=402) of patients with FM was obtained from the Fibromyalgia Association of Aragon, Spain. Patients were assessed with the CPCI-42, the Fibrofatigue Scale (FFS), the EuroQol-5D (EQ-5D), and the Hospital Anxiety and Depression Scale (HADS). The psychometric properties of the CPCI-42 were valid and factor analyses supported the eight-factor structure described in patients with chronic pain. Illness focused coping strategies (i.e., guarding, resting, and asking for assistance) were strongly correlated with each other, positively correlated with disability and depression, and negatively correlated with quality of life, indicating construct validity. Seeking social support was weakly correlated with any other scale or outcome, confirming it belongs to a different group of coping strategies. The wellness-focused group of coping strategies was the most incoherent group. Task persistence correlated with illness-focused strategies and negative outcomes, indicating that it should be included in the illness-focused group. However, other wellness-focused strategies, including relaxation, exercise, and coping self-statements, were correlated with each other, negatively correlated with depression, and positively correlated with quality of life. Future research directions and clinical implications are discussed. PMID- 17400389 TI - Study of the influence of the dielectric layer thickness in a CNT-FED. AB - An experimental investigation is carried out to study the influences of the dielectric layer thickness variation on the field emission characteristics and luminance distribution in a CNT-FED fabricated by screen-printing. Two steps are contained in the investigation: (1) the dielectric layer thickness fluctuations are presented with an ultrasonic thickness gauge, and (2) a simulation model is constructed to study the corresponding influences of the dielectric layer thickness fluctuations on the field emission characteristics and luminance uniformity on the screen. Our findings indicate that the dielectric layer thickness fluctuations are mainly larger than 5 microm, which mean the dielectric layer thickness fluctuation is an important cause of the non-uniform luminance distribution according to the analysis results from our simulation model. From the simulation results, we also determine the tolerance of the dielectric layer thickness in a CNT-FED to achieve uniform luminance and spot size on the screen. PMID- 17400390 TI - Concurrent infection with Trichinella spiralis and other helminths in pigs. AB - The aim of this study was to investigate possible influence of different helmintosis in the development of Trichinella spiralis in experimental infected pigs. Forty-two Iberian pigs were allocated to six groups. Three groups were single inoculated with Ascaris suum, Metastrongylus apri or T. spiralis, respectively. Two groups were co-infected with T. spiralis and A. suum or T. spiralis and M. apri, respectively, while the last group included uninfected control pigs. Clinical signs were only observed in pigs with single or concurrent M. apri infections, with more severe respiratory symptoms in pigs with mixed M. apri infection. The number of A. suum and M. apri lung larvae, intestinal larvae of A. suum and adult M. apri were reduced in pigs with mixed Trichinella infections compared to pigs with single infections. In contrast, the number of liver white spots was higher in pigs with mixed infections. While T. spiralis muscular larval burdens were increased in pigs concomitantly infected with M. apri, they were reduced in pigs concomitantly infected with A. suum, compared to pigs receiving single infections with either of these helminths. Pigs with single or mixed A. suum infections showed higher eosinophil levels compared to the remaining groups. IgGt, IgG1, IgG2 and IgM against T. spiralis antigen could not be detected in pigs with single Ascaris or Metastrongylus infections, indicating that no cross-antibodies were produced. IgGt, IgG1 and IgM antibodies were detected earlier and generally at higher levels in mixed T. spiralis infections compared to single T. spiralis infections. The results suggest that T. spiralis had a low synergistic interaction with M. apri in concomitantly infected pigs, and an antagonistic interaction in concurrent infection with A. suum. PMID- 17400391 TI - Combining the effects of supplementary feeding and copper oxide needles for the control of gastrointestinal nematodes in browsing goats. AB - The aim was to assess the benefits obtained from combining supplementary feeding and copper needles (COWP), compared to the use of both approaches independently, for the control of gastrointestinal nematode (GIN) infections in browsing kids. Forty-four nematode free Criollo kids were exposed to natural parasite infection. The kids were divided into six experimental groups: not treated, supplemented (NT S), not treated, not supplemented (NT-NS), moxidectin treated, supplemented (M S), moxidectin treated not supplemented (M-NS), copper treated, supplemented (COWP-S) and copper treated, non-supplemented (COWP-NS). Copper treated groups received Copinox (2 g capsules) on day 0 and on day 60 of the trial. Moxidectin treated groups received Cydectin (0.2 mg/kg of body weight s.c.) every 28 days. Three of the groups received individual supplementation (100 g of feed/day fresh basis; 74% sorghum: 26% soybean meal; NT-S, M-S and COWP-S) and the other three groups were not supplemented (NT-NS, M-NS and COWP-NS). Animals browsed native vegetation (6.5 h/day) during the wet season (154 days). Kids were weighed every 14 days to determine live weight gain (LWG) and blood and faecal samples were obtained to determine packed cell volume (PCV), haemoglobin (Hb), peripheral eosinophil counts (PEC) and faecal egg counts (FEC). At the end of the trial, four kids of each group were euthanatized (six kids in each COWP treated group). Worm burdens, female worm lengths and prolificacy were determined. Liver samples were used to determine copper concentration and were stained with haematoxylin eosin to determine microscopic lesions. Animals receiving the combination of supplementary feeding and COWP improved their LWG, PCV and Hb to similar levels of animals with suppressive AH treatment. This was not the case when COWP was used without supplementation. Liver copper concentration in COWP treated groups increased significantly especially in the COWP-NS kids but this was not associated with liver lesions or clinical signs. Post-mortem Haemonchus contortus and Trichostrongylus colubriformis worm counts had a tendency to be reduced in the different groups (66-35% reduction) compared to NT-NS group at the end of the trial (P>0.05). Also, COWP treatment and/or supplementation reduced female worm length of T. colubriformis and prolificacy of H. contortus and T. colubriformis. This study, confirmed the value of nutritional supplementation in the control of GIN in growing kids. The use of COWP in addition to supplementation had a limited contribution on the kids' resilience against GIN. This may be due to the reduced infection of H. contortus during this trial. PMID- 17400392 TI - Cultured epithelial autograft (CEA) in burn treatment: three decades later. AB - Methods for handling burn wounds have changed in recent decades and increasingly aggressive surgical approach with early tangential excision and wound closure is being applied. Split-thickness skin (STSG) autografts are the "gold standard" for burn wound closure and remain the mainstay of treatment to provide permanent wound coverage and achieve healing. In some massively burned patients, however, the burns are so extensive that donor site availability is limited. Fortunately, considerable progress has been made in the culture of human keratinocytes and it is now possible to obtain large amounts of cultured epithelium from a small skin biopsy within 3-4 weeks. Questions related to optimal cell type for culture, culture techniques, transplantation of confluent sheets or non-confluent cells, immediate and late final take, carrier and transfer modality, as well as final outcome, ability to generate an epithelium after transplantation, and scar quality are still not fully answered. Progress accomplished since Reinwald and Green first described their keratinocyte culture technique is reviewed. PMID- 17400393 TI - High-dose chemotherapy with autologous stem cell support in first-line treatment of aggressive non-Hodgkin lymphoma - results of a comprehensive meta-analysis. AB - BACKGROUND: Randomized controlled trials (RCTs) reported conflicting results on the impact of high-dose chemotherapy (HDCT) and autologous stem cell transplantation in the first-line treatment of patients with aggressive non Hodgkin lymphoma (NHL). METHODS: We performed a systematic meta-analysis to assess the efficacy HDCT compared to conventional chemotherapy in aggressive NHL patients with regard to complete response (CR), overall survival (OS), event-free survival (EFS), toxicity, and impact of the age-adjusted International Prognostic Index (aaIPI) risk factors. We searched the Cochrane Library, MEDLINE and other databases (1/1990 to 1/2005). Hazard ratio (HR), relative risks (RR) and 95% confidence intervals (CIs) were calculated using the fixed effect model. RESULTS: Fifteen RCTs including 2728 patients were identified. HDCT improved CR when compared to conventional chemotherapy (RR 1.11, CI 1.04-1.18). Overall, there was no evidence for HDCT to improve OS (HR 1.05, 95% CI 0.92-1.19) or EFS (HR 0.92, 95% CI 0.80-1.05) when compared with conventional chemotherapy. However, subgroup analysis indicated OS differences (p=0.032) between good (HR 1.46, 95% CI 1.02 2.09) and poor risk (HR 0.95, 95% CI 0.81-1.11) patients. Conflicting results were reported for poor risk patients, where some studies reported improved and others reduced OS and EFS after HDCT. CONCLUSION: There was no evidence that HDCT improved OS and EFS in good risk NHL patients. The evidence for poor risk patients is inconclusive. HDCT should not be further investigated in good risk patients with aggressive NHL but high quality studies in poor risk patients are warranted. PMID- 17400394 TI - Genome instability and DNA repair in brain, ageing and neurological disease. PMID- 17400395 TI - Second to fourth digit length ratio (2D:4D) and adult sex hormone levels: new data and a meta-analytic review. AB - The relative length of the second (index) to the fourth (ring) finger (2D:4D) is a putative negative correlate of prenatal testosterone (T) exposure. Therefore, 2D:4D (and to a lesser extent D(r-l), the difference between 2D:4D in the right hand and in the left hand) has often been used to study effects of prenatal androgenization on human behavior and cognition. However, evidence suggests that 2D:4D may also be related to levels of circulating sex hormones in adults. This would question the validity of 2D:4D as a means of studying the effects of prenatal sex hormones. Here we present new data from two non-clinical samples (64 women and 102 men) regarding the relationships of 2D:4D and D(r-l) with circulating sex hormone levels. We then present a meta-analytic review of all the present evidence regarding this issue. The results suggest that, in the normal population, 2D:4D and D(r-l) are not associated with adult sex hormone levels. The findings from this current study add to the growing body of evidence demonstrating that 2D:4D is a suitable tool to study the effects of prenatal androgenization on human behavior and cognition. PMID- 17400396 TI - Minor beta-thalassemia as a predisposing factor for preeclampsia in pregnant women. PMID- 17400397 TI - Demineralized bone matrix enriched with human recombinant interleukin-11: a novel therapeutic option in treatment of delayed unions and nonunions? PMID- 17400398 TI - Is atherosclerosis a neurogenic phenomenon? AB - Identified risk factors for atherosclerosis include diet, age, gender, family history, stress, lifestyle, smoking, diabetes, dyslipidemias, hypertension, and HIV. The mechanistic rationale to explain these associations remains poorly understood. We believe that these seemingly unrelated entities may promote atherosclerosis through a common pathway by inducing adventitial autonomic dysfunction, specifically as an adventitial stress dysfunction of neurogenic origin. Atherosclerosis may represent a local vascular manifestation of the global autonomic dysfunction induced by age, smoking, hypertension, HIV, and diabetes. Atherosclerosis may also participate in a feed-forward cycle as aging, diabetes, dyslipidemia, and hypertension may also represent independent downstream consequences of global sympathetic bias. Chronic physiologic stress and behavioral stress can shift the autonomic balance towards a state of sympathetic predominance. The highly communicable nature of behavioral stress may partially implicate the familial association of atherosclerosis as an epigenetic phenomenon, independent of putative genetic mechanisms. Host stress, global autonomic dysfunction, and sympathetic bias may also arise from chronic maladaptive consumption of stressed foods, as organisms detect and assimilate the stress phenotypes of their dietary constituents through a process called xenohormesis. The benefits of exercise may operate through reduction of chronic physiologic stress associated with global sympathetic bias. The neurogenic adventitial stress response may explain the local tissue remodeling seen in atherosclerosis, including adventitial adipose dysfunction, inflammation, adventitial angiogenesis, thrombosis, and endothelial dysfunction. We believe that the locations of atherosclerotic lesions correspond to regions of neurogenic adventitial autonomic dysfunction, in similar fashion to the segmental patterns of involvement found in inflammatory bowel disease. The diffuse atherosclerosis exhibited in transplanted hearts may reflect a diffuse sympathetic bias of the donor heart, since tissues and organs exhibit an intrinsic sympathetic bias in the absence of an extrinsic source of autonomic hegemony. Once we regard atherosclerosis as a neurogenic phenomenon manifested in adventitial autonomic dysfunction, novel diagnostic and therapeutic paradigms become evident. PMID- 17400399 TI - Intestinal occlusion caused by Rosai-Dorfman disease mimicking colonic diverticulitis. AB - Rosai-Dorfman disease (RDD) involves the gastrointestinal tract only in exceptional cases, and this very unusual site of presentation can confuse the pathologist. We present a case of RDD manifesting as an intestinal occlusion caused by colonic diverticulitis. The patient was a 79-year-old man with myelodysplasia, who presented with fever, abdominal pain, and constipation. Colonoscopy revealed sigmoiditis and diverticulosis. Microscopic study of the sigmoid colon surgical specimen showed the histological and immunological features of RDD. No human DNA of herpesvirus types 6 and 8 (HHV6/HHV8), Epstein Barr virus (EBV), and cytomegalovirus (CMV) was detected in tissue by polymerase chain reaction. Electron microscopic study revealed no microbes or viral particles. Widespread nodal and extranodal RDD occurred, and the patient died 2y after initial surgery. The etiology of RDD is still under debate. We discuss the association of RDD with hematological disorders. PMID- 17400400 TI - Glassy cell carcinoma of the endometrium: a case report and review of the literature. AB - Glassy cell carcinomas are composed of malignant cells showing a "ground glass" cytoplasm, distinct cell membranes, and large nuclei with prominent nucleoli. To our knowledge, only 12 cases of glassy cell endometrial carcinomas (EGCC) have been reported until now. A 63-year-old patient complaining of irregular vaginal bleeding underwent hysteroscopy-guided biopsy revealing a well-differentiated endometrial endometrioid adenocarcinoma. The patient underwent left salpingo oophorectomy, total abdominal hysterectomy, and pelvic lymphadenectomy. The final diagnosis was FIGO stage IB poorly differentiated endometrial adenosquamous carcinoma with > 90% of glassy tumor cells. The patient is alive, with no evidence of disease for 69 months after diagnosis. We describe an additional case of EGCC and review the data of the literature, emphasizing the need to strictly define the criteria for the diagnosis and the potential usefulness of assessing biologic parameters for the prognostic characterization of this rare entity. PMID- 17400401 TI - [The prosthetic kits in the prolapse surgery: is it a gadget?]. AB - OBJECTIVE: To search if the prosthetic kits bring an interest, other that financier. MATERIALS AND METHOD: Authors have counted 5 firms presenting the varied kits. The Prolift kit with synthetic pre- cut mesh made of polypropylene, standardized needle, cannulas and protective devices of recovery of mesh arms in 3 versions, anterior, posterior and total. Perigee and Apogee systems in a synthetic version (IntePro) and a biological version (InteXen ++LP) matched of specific needles for the different obturator passages and infra coccygeus. The Avaulta kit declines in a biosynthetic version and a hybrid version with a specific needle for its anterior kit and an other for its posterior kit. The Biomesh Soft system presents a polypropylene mesh posed on a multi pattern sheet to carve following dots with a share of 3 needles according to the type of arm passage ended by a recuperator thread. The Nazca POP Repair System, in its anterior version claims to be able to correct in the same time a urinary incontinence by pre-pubic arms. The polypropylene mesh is perforated and is had with the help a needle for its anterior and posterior kit. Authors have been interested in the research of studies on prosthesis with and without kit. RESULTS: The Prolift kit contains the alone prosthesis having been evaluated without (TVM) and with kit. Authors remind results of two studies, understanding 684 cases without kit and 110 cases with kit. The rate of early per and postoperative complications has been decreased half, the rate of mesh exposure decreased from 11,3 to 4,7%. Studies on Perigee and Apogee IntePro kits, whose meshe and the procedure are similar to the Prolift seem to obtain from results equal. The Biological version has not made the object of study. For the Avauta kit, alone the mesh in plate not pre-cut has been well studied as for its tolerance for the biosynthetic version. Nothing on the hybrid version. No study is found for the Biomesh Soft kit and Nazca POP repair system. DISCUSSION: The cost of these kits, varying 1 to 4, is to take in consideration and to put in scale with the returned service. Alone TVM/Prolift allows to advance in a reply. The utilization of needles, presents in all the kits, but especially devices facilitating the passage and the recovery of prosthesis arms, presents solely in the Prolift kit, reduce the utilization of valves and the risk of tear tissues during their job. The improvement of techniques by elements of the kit make that the surgical procedure become mini invasive. CONCLUSION: The full kits allow a best security of procedures, facilitate the surgical gesture and limit complications. It is necessary to remain vigilant in the evaluation of these new materials "ready to wear" and to require pre clinic and clinic studies before their distribution. PMID- 17400402 TI - [Sentinel node detection applied to breast cancer: 2007 update]. AB - The technique of detection and resection of the sentinel lymph node applied to early breast cancer management aims to spare the patient with a low risk of lymph node involvement an unnecessary axillary lymphadenectomy. This innovating technique lies on the double hypothesis of an accuracy to predict non sentinel lymph node status and to induce a lower morbidity when compared with axillary lymphadenectomy. This multidisciplinary technique depends on surgeons, nuclear physicians and pathologists. In practice sentinel lymph nodes are detected thanks to two types of tracers, the Blue and the colloids marked with technetium, harvested by the surgeon guided by the blue lymphatic channel and the use of a gamma probe detection, analyzed by the pathologist according to a particular procedure with the concept of serial slices, and possibly immuno histo chemistry. The objectives of this review are to specify the state of knowledge concerning the different steps: detection, surgical resection and the pathological analysis of the sentinels lymph nodes and to focus on validated and controversial indications, and on the main ongoing trials. PMID- 17400403 TI - Equine learning behaviour. AB - Scientists and equestrians continually seek to achieve a clearer understanding of equine learning behaviour and its implications for training. Behavioural and learning processes in the horse are likely to influence not only equine athletic success but also the usefulness of the horse as a domesticated species. However given the status and commercial importance of the animal, equine learning behaviour has received only limited investigation. Indeed most experimental studies on equine cognitive function to date have addressed behaviour, learning and conceptualization processes at a moderately basic cognitive level compared to studies in other species. It is however, likely that the horses with the greatest ability to learn and form/understand concepts are those, which are better equipped to succeed in terms of the human-horse relationship and the contemporary training environment. Within equitation generally, interpretation of the behavioural processes and training of the desired responses in the horse are normally attempted using negative reinforcement strategies. On the other hand, experimental designs to actually induce and/or measure equine learning rely almost exclusively on primary positive reinforcement regimes. Employing two such different approaches may complicate interpretation and lead to difficulties in identifying problematic or undesirable behaviours in the horse. The visual system provides the horse with direct access to immediate environmental stimuli that affect behaviour but vision in the horse is of yet not fully investigated or understood. Further investigations of the equine visual system will benefit our understanding of equine perception, cognitive function and the subsequent link with learning and training. More detailed comparative investigations of feral or free-ranging and domestic horses may provide useful evidence of attention, stress and motivational issues affecting behavioural and learning processes in the horse. The challenge for scientists is, as always, to design and commission experiments that will investigate and provide insight into these processes in a manner that withstands scientific scrutiny. PMID- 17400404 TI - Can studies of cognitive abilities and of life in the wild really help us to understand equine learning? PMID- 17400405 TI - Extending the diversity of cytochrome P450 enzymes by DNA family shuffling. AB - The cytochrome P450 enzymes involved in xenobiotic metabolism are an excellent starting point for the directed evolution of novel biocatalysts due to their wide substrate specificity. A shuffled library of three highly homologous mammalian genes (for P450 2C9, P450 2C11 and P450 2C19) was constructed by applying a modified DNA family shuffling procedure. The modifications made to the traditional DNA shuffling protocols involved non-random digestion via the use of different combinations of restriction enzymes (REs) followed by isolation of fragments under 300 bp by size-selective filtration. Shuffled cytochrome P450 mutants were co-expressed in Escherichia coli with their redox partner, NADPH cytochrome P450 reductase (NPR). We report here how non-random fragmentation may help in chimeragenesis within the areas of low sequence similarity such as substrate recognition sites (SRSs) that are generally underrepresented in recombination using the random fragmentation process. Size-selective filtration was used to limit recovery of incompletely digested fragments and consequently minimize the chances for contamination of the shuffled library with parental forms. No parental forms could be detected in the shuffled library using restriction fragment length polymorphism (RFLP) analysis, suggesting the library was free of parental contamination. Sequencing of randomly selected mutants demonstrated a high level of chimeragenesis with on average of 8.0+/-2.2 crossovers and a low level of mutagenesis with 5.2+/-2.8 spontaneous mutations per approximately 1.5 kbp of the full-length P450 sequence. The proportion of properly folded protein as indicated by the observation of characteristic Fe(II).CO vs. Fe(II) difference spectra was 15% (4/27) of analysed mutants. Screening of the shuffled library for indole oxidation revealed four clones with similar or higher levels of indigo pigment production to those of the parental P450s and two clones with elevated P450 expression. In this paper we present a method for the effective family shuffling of cytochrome P450 enzymes, applicable to the creation of mutant libraries with expanded metabolic diversity and with a significant proportion of functional clones. PMID- 17400406 TI - A study of biochemical and functional interactions of Htl1p, a putative component of the Saccharomyces cerevisiae, Rsc chromatin-remodeling complex. AB - HTL1, a small gene of Saccharomyces cerevisiae, encodes a 78-aminoacid peptide that influences the performance of a wide range of cellular processes [Lanzuolo, C., Ederle, S., Pollice, A., Russo, F., Storlazzi, A., Pulitzer, J.F., 2001. The HTL1 gene,YCR020W-b of Saccharomyces cerevisiae is necessary for growth at 37 degrees C, and for the conservation of chromosome stability and fertility. Yeast, 18, 1317-1330]. Genetic interactions and co-immunoprecipitation experiments indicate a role for Htl1p in functions controlled by RSC, a multiprotein, ATP dependent, chromatin-remodeling complex [Lu, Y.M., Lin, Y.R., Tsai, A., Hsao, Y.S., Li, C.C., Cheng, M.Y., 2003. Dissecting the pet18 mutation in Saccharomyces cerevisiae: HTL1 encodes a 7-kDa polypeptide that interacts with components of the RSC complex. Mol. Genet. Genomics., 269, 321-330] [Romeo, M.J., Angus-Hill, M.L., Sobering, A.K., Kamada, Y., Cairns, B.R., Levin, D.E., 2002. HTL1 encodes a novel factor that interacts with the Rsc chromatin-remodeling complex in Saccharomyces cerevisiae. Mol. Cell. Biol., 22, 8165-8174]. Htl1p and RSC components, share the property of associating with TBP a component of general multiprotein transcription factor TFIID [Sanders, S.L., Jennings, J., Canutescu, A., Link, A.J., Weil, P.A., 2002. Proteomics of the eukaryotic transcription machinery: identification of proteins associated with components of yeast TFIID by multidimensional mass spectrometry. Mol. Cell. Biol. 22, 4723-4738]. We confirm, by integrating genetic and biochemical experiments, that Htl1p binding to the RSC complex is direct and physiologically relevant and show that it is mediated by Rsc8p, a core component of the RSC complex. Deletion of HTL1, like depletion of RSC core subunits [Moreira, J.M., Holmberg, S., 1999. Transcriptional repression of the yeast CHA1 gene requires the chromatin remodeling complex Rsc. Embo J., 18, 2836-2844], leads to constitutive transcription of the CHA1 locus. This transcriptional phenotype exhibits variable penetrance. Deletion of HTL1 also leads to hydroxyurea hypersensitivity at 30 degrees C, suggesting a defect in replication/repair. This defect leads, during cell growth, to selection of mutations at the SIR3 locus that suppress hydroxyurea sensitivity. PMID- 17400407 TI - Intron distribution in Plantae: 500 million years of stasis during land plant evolution. AB - Little is known about the evolution of the intron-exon organization in the more primitive groups of land plants, and the intron distribution among Plantae (glauco-, rhodo-, chloro- and streptophytes) has not been investigated so far. The present study is focused on some key species such as the liverwort Marchantia polymorpha, representing the most ancient lineage of land plants, and the streptophycean green alga Mesostigma viride, branching prior to charophycean green algae and terrestrial plants. The intron distribution of six genes for sugar phosphate metabolism was analyzed including four different glyceraldehyde-3 phosphate dehydrogenases (GAPDH), the sedoheptulose-1,7-bisphosphatase (SBP) and the glucose-6-phosphate isomerase (GPI). We established 15 new sequences including three cDNA and twelve genomic clones with up to 24 introns per gene, which were identified in the GPI of Marchantia. The intron patterns of all six genes are completely conserved among seed plants, lycopods, mosses and even liverworts. This intron stasis without any gain of novel introns seem to last for nearly 500 million years and may be characteristic for land plants in general. Some unique intron positions in Mesostigma document that a uniform distribution is no common trait of all streptophytes, but it may correlate with the transition to terrestrial habitats. However, the respective genes of chlorophycean green algae display largely different patterns, thus indicating at least one phase of massive intron rearrangement in the green lineage. We moreover included rhodophyte and glaucophyte reference sequences in our analyses and, even if the well documented monophyly of Plantae is not reflected by a uniform intron distribution, at least one GPI intron is strictly conserved for 1.5 billion years. PMID- 17400408 TI - Genotypic versus phenotypic identification of staphylococcal species of canine origin with special reference to Staphylococcus schleiferi subsp. coagulans. AB - A comparative study was performed to examine the respective accuracy of 16S rDNA sequencing and of the commercial biochemical assay ID32 STAPH (bioMerieux, Marcy l'Etoile, France) in the identification of 232 staphylococcal samples representing 20 species and subspecies isolated from 367 dogs. Notable differences in species distribution were observed by comparing genotypic and phenotypic data. Partial sequencing of 16S rDNA resulted in an unambiguous identification of 226 (97.4%) of the isolates, whereas the phenotypic approach resulted in a correct diagnosis of 162 (69.8%) of the isolates. Statistical agreement between genotypic and phenotypic identification of staphylococci was substantial (Kappa coefficient of 0.6-0.8) for Staphylococcus aureus, S. hominis, S. warneri, S. cohnii subsp. urealyticus, and S. simulans, and "almost perfect" (Kappa coefficient of 0.8-1) for S. intermedius, S. epidermidis, S. equorum, S. haemolyticus, S. sciuri, and S. kloosi. No agreement above that expected by chance (Kappa coefficient=0) was observed for S. schleiferi subsp. coagulans, which was either confounded with S. intermedius and S. capitis, or categorized as unacceptable by the biochemical assay. Given the growing importance of this pathogen in veterinary medicine and its frequent misidentification with related staphylococci, a PCR-RFLP approach producing a S. schleiferi-specific restriction profile was developed. This fast and reliable assay represents a valuable tool in assisting in the monitoring of this pathogen. PMID- 17400409 TI - Characterization of multidrug resistant Salmonella recovered from diseased animals. AB - Three hundred and eighty Salmonella isolates recovered from animal diagnostic samples obtained from four state veterinary diagnostic laboratories (AZ, NC, MO, and TN) between 2002 and 2003 were tested for antimicrobial susceptibilities and further characterized for bla(CMY) beta-lactamase genes, class 1 integrons and genetic relatedness using PFGE. Forty-seven serovars were identified, the most common being S. Typhimurium (26%), S. Heidelberg (9%), S, Dublin (8%), S. Newport (8%), S. Derby (7%), and S. Choleraesuis (7%). Three hundred and thirteen (82%) isolates were resistant to at least one antimicrobial, and 265 (70%) to three or more antimicrobials. Resistance was most often observed to tetracycline (78%), followed by streptomycin (73%), sulfamethoxazole (68%), and ampicillin (54%), and to a lesser extent chloramphenicol (37%), kanamycin (37%), amoxicillin-clavulanic acid (20%), and ceftiofur (17%). With regards to animal of origin, swine Salmonella isolates displayed the highest rate of resistance, being resistant to at least one antimicrobial (92%), followed by those recovered from turkey (91%), cattle (77%), chicken (68%), and equine (20%). Serovars commonly showing multidrug resistance (MDR) to > or =9 antimicrobials were S. Uganda (100%), S. Agona (79%), and S. Newport (62%), compared to S. Heidelberg (11%) and S. Typhimurium (7%). Class-1 integrons were detected in 43% of all isolates, and were found to contain aadA, aadB, dhfr, cmlA and sat1 gene cassettes alone or in various combinations. All ceftiofur resistant isolates (n=66) carried the bla(CMY) beta-lactamase gene. A total of 230 PFGE patterns were generated among the 380 isolates tested using XbaI, indicating extensive genetic diversity across recovered Salmonella serovars, however, several MDR clones were repeatedly recovered from different diseased animals. PMID- 17400410 TI - Combined effects of mycotoxin mixtures on human T cell function. AB - Combined effects of mycotoxin mixtures are of major concern due to the widespread occurrence of human exposure to mixtures. Owing to the lack of sufficient mixture data regarding the human immune function, the aim of this study was to specify, evaluate and predict the combined effects of mycotoxin mixtures upon the functional activity of immune cells. Therefore, a well established model for immunotoxic studies using stimulated human peripheral blood mononuclear cells (PBMC) was applied. To predict the combined effect of the mixture the concepts of concentration addition and response addition were used. Comparable to the individual mycotoxins the mixture suppressed the cytokine production in a concentration-dependent manner. The mixture effect was stronger than the effects caused by the toxins applied singly. The response addition concept described the mixture data within their experimental uncertainty. The in vitro test model presented here was practical to detect and quantify combined effects. Low and weak effect concentrations of mycotoxins may cause strong inhibitory effects on immune functions when occurring together. The experimental design using the concepts of concentration addition and response addition was suitable for predicting the combined effects of mycotoxin mixtures in functional in vitro assays. PMID- 17400412 TI - Altered default mode network activity in patient with anxiety disorders: an fMRI study. AB - Anxiety disorder, a common mental disorder in our clinical practice, is characterized by unprovoked anxiety. Medial prefrontal cortex (MPFC) and posterior cingulate cortex (PCC), which closely involved in emotional processing, are critical regions in the default mode network. We used functional magnetic resonance imaging (fMRI) to investigate whether default mode network activity is altered in patients with anxiety disorder. Ten anxiety patients and 10 healthy controls underwent fMRI while listening to emotionally neutral words alternating with rest (Experiment 1) and threat-related words alternating with emotionally neutral words (Experiment 2). In Experiment 1, regions of deactivation were observed in patients and controls. In Experiment 2, regions of deactivation were observed only in patients. The observed deactivation patterns in the two experiments, which included MPFC, PCC, and inferior parietal cortex, were similar and consistent with the default model network. Less deactivation in MPFC and greater deactivation in PCC were observed for patients group comparing to controls in Experiment 1. Our observations suggest that the default model network is altered in anxiety patients and dysfunction in MPFC and PCC may play an important role in anxiety psychopathology. PMID- 17400411 TI - Hypotonic swelling of salicylate-treated cochlear outer hair cells. AB - The outer hair cell (OHC) is a hydrostat with a low hydraulic conductivity of Pf=3x10(-4) cm/s across the plasma membrane (PM) and subsurface cisterna that make up the OHC's lateral wall. The SSC is structurally and functionally a transport barrier in normal cells that is known to be disrupted by salicylate. The effect of sodium salicylate on Pf is determined from osmotic experiments in which isolated, control and salicylate-treated OHCs were exposed to hypotonic solutions in a constant flow chamber. The value of Pf=3.5+/-0.5x10(-4) cm/s (mean+/-s.e.m., n=34) for salicylate-treated OHCs was not significantly different from Pf=2.4+/-0.3x10(-4) cm/s (mean+/-s.e.m., n=31) for untreated OHCs (p=.3302). Thus Pf is determined by the PM and is unaffected by salicylate treatment. The ratio of longitudinal strain to radial strain epsilonz/epsilonc=-0.76 for salicylate-treated OHCs was significantly smaller (p=.0143) from -0.72 for untreated OHCs, and is also independent of the magnitude of the applied osmotic challenge. Salicylate-treated OHCs took longer to attain a steady-state volume which is larger than that for untreated OHCs and increased in volume by 8-15% prior to hypotonic perfusion unlike sodium alpha-ketoglutarate-treated OHCs. It is suggested that depolymerization of cytoskeletal proteins and/or glycogen may be responsible for the large volume increase in salicylate-treated OHCs as well as the different responses to different modes of application of the hypotonic solution. PMID- 17400413 TI - Decision-making at menopause: a randomized controlled trial of a computer-based hormone therapy decision-aid. AB - OBJECTIVE: Decision-making at menopause remains a challenge for women and their health care providers as the paradigm for hormone therapy continues to evolve. The role of decision-support for this process remains to be defined. METHODS: A randomized controlled trial of a computer-based hormone therapy (HT) decision-aid versus a control intervention consisting of a printed pamphlet among 177 post menopausal women receiving care in a Veterans Affairs Medical Center. RESULTS: Participants found the computer-based decision-aid easy to use and retained risk information incorporated from emerging scientific data. There was no difference between groups with respect to the primary outcomes of knowledge, satisfaction with decision, decisional conflict or HT use. A trend was reported towards decreased decisional conflict in the evidence in decision-making (p=0.07) and factors of uncertainty (p=0.06) domains among the subset of participants who were on HT at baseline and used the computer-based decision-aid. CONCLUSION: The computer-based decision-aid was able to effectively incorporate emerging scientific information but was no more effective than a printed pamphlet control with regard to improving decision-process outcomes. PRACTICE IMPLICATIONS: The incremental benefit of a complex versus simpler decision-aid for post-menopausal women remains to be established prior to widespread dissemination of interactive computer-based HT decision-aids. PMID- 17400414 TI - Treatment completion on an inpatient detoxification unit: impact of a change to sublingual buprenorphine-naloxone. AB - PURPOSE: Buprenorphine is commonly used for opioid detoxification. The goal of this study was to determine whether a change from the intramuscular (IM) buprenorphine to the sublingual (SL) formulation of buprenorphine-naloxone was associated with improved treatment completion rates on an inpatient detoxification unit. METHODS: This study was conducted at the Johns Hopkins Bayview Medical Center (JHBMC) Chemical Dependence Unit (CDU), a 26-bed, 3-day inpatient detoxification unit providing detoxification from opioids, alcohol, and sedatives. The opioid detoxification protocol was changed from IM buprenorphine (0.3 mg bid for 3 days) to SL buprenorphine-naloxone (8, 8, and 6 mg on sequential days, plus 2 mg on the morning of discharge). For the 3 months prior to and after the change in protocol, data were collected retrospectively on demographics, type of dependence being treated, and type of discharge. FINDINGS: A total of 1,168 patients were admitted to the JHBMC CDU during the period studied. In the 3 months prior to the change in buprenorphine protocol, 353 of 483 patients admitted for treatment of opioid dependence (73.1%) completed treatment, compared with 407 of 473 patients admitted after the change (86.0%); this difference was highly significant (p < .0001). Among 212 patients who did not receive treatment for opioid dependence over the same period, the rates of treatment completion did not change significantly (89.8% before vs. 83.0% after; p = .208). CONCLUSIONS: A change from IM buprenorphine to SL buprenorphine naloxone for opioid detoxification was associated with a significant improvement in completion rates at this inpatient treatment program. PMID- 17400415 TI - Comparison of demographic and clinical characteristics between opioid-dependent individuals admitted to a community-based treatment setting and those enrolled in a research-based treatment setting. AB - Despite the significant developments in pharmacotherapy and behavioral treatments for addiction, the dissemination of new treatment methods into the community has been slow. It has been pointed out that treatments developed in research settings may be impractical in community treatment settings, which might help explain the transition lag. Screening and recruitment of participants for research studies might partially explain this, as there is evidence that substance-abusing individuals who participate in clinical research are different on a number of measures from treatment seekers. However, no study has directly compared treatment seekers with research participants drawn from similar populations using prospective methods. This study compared the demographic characteristics, drug use and psychosocial problem severity levels, and personality traits of opioid dependent individuals seeking help in a community setting (n = 502) with those of opioid-dependent individuals in a primarily research-based drug abuse treatment setting (n = 459); both settings offered a similar set of treatment services (opioid agonist medication and counseling). Although the overall findings revealed numerous similarities between the groups, differences were also observed. Most notably, there were significantly fewer women in the research sample than in the community-based treatment sample. Other differences included a modest but statistically significant increase in psychosocial problem severity levels in the community-based treatment sample and higher drug use problem severity levels in the research sample. Interestingly, many of these differences were strongest in women as compared with men. PMID- 17400416 TI - Veterans Affairs facility performance on Washington Circle indicators and casemix adjusted effectiveness. AB - Self-administered Addiction Severity Index (ASI) data were collected on 5,723 patients who received substance abuse treatment in 1 of 110 programs located at 73 Veterans Affairs facilities. The associations between each of three Washington Circle (WC) performance indicator scores (identification, initiation, and engagement) and their casemix-adjusted facility-level improvement in ASI drug and alcohol composites 7 months after intake were estimated. Higher initiation rates were not associated with facility-level improvement in ASI alcohol composite scores but were modestly associated with greater improvements in ASI drug composite scores. Identification and engagement rates were unrelated to 7-month outcomes. WC indicators focused on the early stages of treatment may tap necessary but insufficient processes for patients with substance use disorder to achieve good posttreatment outcomes. Ideally, the WC indicators would be supplemented with other measures of treatment quality. PMID- 17400417 TI - Prognosis and postoperative lymphocyte count in patients with hepatocellular carcinoma who received intraoperative allogenic blood transfusion: a retrospective study. AB - AIMS: The effect of perioperative blood transfusion on the survival of hepatocellular carcinoma (HCC) has not been fully investigated. To clarify the prognostic value of intraoperative allogenic blood transfusion, we conducted a comparative retrospective analysis of 224 patients with HCC who underwent hepatic resection. METHODS: We compared clinicopathologic background and survival after hepatic resection between patients who received intraoperative blood transfusion (n=101) and those who did not (n=123). RESULTS: Patients with blood transfusion had a larger tumor and more frequent vascular invasion than those without blood transfusion. The 5-year cancer-related survival rate after hepatic resection, but not the disease-free survival rate, was significantly lower in patients who underwent blood transfusion than in those who did not (38.3% vs. 66.7%, P<0.01). Multivariate analysis showed intraoperative blood transfusion (P=0.02), microscopic portal invasion (P<0.01), and preoperative serum alpha-fetoprotein elevation (P=0.03) to be independent risk factors for poor outcome after hepatic resection. The negative effect of blood transfusion on postoperative survival was observed only in patients with a tumor larger than 50mm in diameter. The absolute peripheral blood lymphocyte count on postoperative day 1 was significantly lower in patients who underwent blood transfusion (880/mm(3)) than in those who did not (1081/mm(3)) (P<0.01). CONCLUSIONS: Our data suggest that intraoperative blood transfusion results in immunosuppression in the early postoperative period, allowing for progression of residual HCC after resection. Therefore, intraoperative allogenic blood transfusion should be avoided in patients with resectable HCC, particularly in those with a large tumor. PMID- 17400418 TI - Strategies for the treatment of synchronous liver metastasis. AB - AIM: Observe the outcomes after complete simultaneous or delayed resection of synchronous liver metastasis (SLM) from colorectal cancer (CRC). METHODS: From 1994 to 2005, 119 patients were diagnosed with CRC and SLM; 57 patients had simultaneous resection (group I) and 62 patients had staged resection (group II). Perioperative chemotherapy was considered completed if all expected cycle were administrated. RESULTS: Overall survival rates of group I-group II at 1, 3 and 5 years were respectively 91%-93% (p=0,3), 59%-57% (p=0,09) and 32%-25% (p=0,06). The median survival time of group I-group II were respectively 46 months-40 months (p=0,07). There was no statistical difference on survival regarding location of metastasis (p=0,09) or primary tumor location (p=0,2). Patients with simultaneous or staged resection receiving optimal treatment (R0 liver surgery and complete chemotherapy) were respectively 89% and 67% (p=0,04). Twenty three patients developed isolated liver recurrence with higher frequency in staged patients (26% vs 9% p=0,03) without impairment of survival. CONCLUSIONS: Because of postoperative morbidity and prolonged tiring treatment, many patients having staged resection were under treated. However we did not observe statistical difference on survival but we supported that simultaneous resection has to be prefer to achieve an optimal treatment. Lung and bone metastasis are the new challenge for oncologists. PMID- 17400419 TI - Evaluation of VEGF A expression and microvascular density as prognostic factors in extrahepatic cholangiocarcinoma. AB - OBJECTIVES: Angiogenesis is essential for tumor growth and metastasis. An association between microvessel density, a measure of tumor angiogenesis, and conventional prognostic variables has been shown for many different tumor entities. In extrahepatic cholangiocarcinoma, the VEGF expression and microvessel density have rarely been investigated. METHODS: Paraffin-embedded specimens from 51 resected adenocarcinomas of the extrahepatic bile duct were immunostained for vascular endothelial growth factor A (VEGF A) and CD 34 to evaluate the microvessel density (MVD). VEGF A staining was evaluated by combining intensity and percentage of positive tumor cells, as low (expression equal or below the median), or high (above the median). Microvessel density was assessed using a method published by Weidner et al. RESULTS: Median disease free survival (DFS) of the study group was 12.5 months (range, 1-66.3 months). DFS was calculated in the 39 patients with complete resection. It was significantly better in patients with low microvessel density than DFS in patients with high microvessel density (33 months (range, 3-66.3 months) vs. 21.8 months (range, 1.6-31.6 months); p=0.022). In contrast, VEGF A expression did not correlate with survival. There was a trend toward a higher VEGF A expression in highly vascularized tumors (p=0.08), but failed to reach statistic significance. CONCLUSIONS: The present study indicates, that vascularisation has an important impact on survival of extrahepatic cholangiocarcinoma patients. Other molecules than VEGF A are probably involved in neovascularization in extrahepatic cholangiocarcinoma. PMID- 17400420 TI - A population-based study of radiotherapy in a cohort of patients with rectal cancer diagnosed between 1996 and 2000. AB - AIMS: To study, in a population-based setting, the use of delayed radiotherapy (RT) in a cohort of 2008 unselected rectal cancer patients diagnosed between 1996 and 2000. PATIENTS AND METHODS: Radiation within 6 months of diagnosis was considered part of the primary treatment (PRT). RT given 6 months or later after diagnosis or after PRT was considered as delayed or secondary RT (SRT). Number, percentage and cumulative proportion of patients receiving SRT were calculated. The odds for receiving SRT (total and for recurrent rectal cancer only) were studied by logistic regression analysis, taking into account age, gender, co morbidity, socio-economic status, stage, prior PRT and RT department (2 departments, each serving general hospitals only). RESULTS: Forty-six percent of all newly diagnosed patients received RT. Ten percent (n=203) received at least once SRT, either after PRT or as first RT, of which 96 patients for a relapsed rectal tumour (31 after PRT on the rectal tumour, 65 as a first radiation treatment). In a multivariate analysis of patients with rectal recurrence secondary pelvic irradiation was less often given after primary irradiation (OR: 0.7, 95% CI: 0.4-1.1). Patients with a stage III significantly more often received SRT on a recurrence (OR=2.5, 95% CI=1.4-4.5). Generally, patients in the eastern department received more often PRT and less often SRT for recurrence (OR: 0.5, 95% CI: 0.3-0.8). CONCLUSIONS: Five percent of all patients with rectal cancer received SRT on a recurrent tumour, with a large variation between the two RT departments in the region. PMID- 17400421 TI - Improving laser-induced thermotherapy of liver metastases--effects of arterial microembolization and complete blood flow occlusion. AB - INTRODUCTION: A prerequisite for an oncologically curative application of laser induced thermotherapy (LITT) of liver metastases is complete tumor destruction. This increased effectiveness was achieved experimentally by combining LITT with interrupted hepatic perfusion. The aim of this study was to evaluate whether an interventional selective arterial microembolization might be as effective as complete blood flow occlusion using an open Pringle's maneuver. PATIENTS AND METHODS: We included patients with unresectable colorectal liver metastases. LITT was performed without interrupted hepatic perfusion (control group) compared to LITT in combination with interrupted perfusion either by embolization of intraarterial degradable starch microspheres (DSM) (percutaneous access) or by complete hepatic inflow occlusion (Pringle's maneuver; open access). Online monitoring was performed using intraoperative ultrasound or MRI. Volumetric techniques were used to assess metastases and postinterventional lesions. RESULTS: Fifty-six patients with 104 metastases (control group (25), DSM (37), and Pringle (42)) were treated. The preinterventional tumor volumes were significantly smaller than the postinterventional lesion volumes (control group: 9.8 vs. 25.3 cm3; DSM: 9.5 vs. 65.4 cm3; Pringle: 12.9 vs. 76.5 cm3). The morbidity rate was 21.4% without treatment-related mortalities. After 6 months follow-up, tumor recurrence was diagnosed in 6 patients (control group (4), LITT with DSM (1), and Pringle (1)). CONCLUSIONS: Combining LITT with blood flow occlusion leads to a significant increase in lesion size. The application of DSM offers a safe and effective alternative to the open access with Pringle's maneuver. Compared to LITT-monotherapy, this modality achieves significantly larger thermal lesions with the need of fewer applications. PMID- 17400422 TI - Hepatic artery infusion of high-dose melphalan at reduced flow during isolated hepatic perfusion for the treatment of colorectal metastases confined to the liver: a clinical and pharmacologic evaluation. AB - Isolated hepatic perfusion (IHP) offers the advantage of high local drug exposure with limited systemic toxicity. To increase local drug exposure, we administered melphalan at a reduced flow in the hepatic artery during IHP (hepatic artery infusion, hepatic artery-portal vein perfusion, HI-HPP). Between December 2001 and December 2004, 30 patients with colorectal cancer liver metastases underwent HI-HPP with 200mg melphalan. Samples of the perfusate were taken for pharmacokinetic analysis. Patients were monitored for response, toxicity and survival. Perfusion was aborted prematurely in 2 patients due to leakage. During melphalan administration in the hepatic inflow cannula a mean flow rate of 121.3 mL/min and mean pressure of 62.5mm Hg were achieved. One patient died within 30 days after HI-HPP. Four patients developed veno-occlusive disease (VOD), while 2 patients showed signs of VOD. Twelve patients showed hepatic response, with a median duration of response of 11.5 months, according to WHO criteria. Although HI-HPP results in high perfusate melphalan concentration levels, it is associated with a relatively high level of hepatotoxicity and a limited response rate. We believe that the low flow and pressure rates found in this study can result in reduced drug penetration of the tumour and thus limited tumour response. PMID- 17400423 TI - Prognostic and predictive factors after surgical treatment for locally recurrent rectal cancer: a single institute experience. AB - OBJECTIVE: Resection of locally recurrent rectal cancer (LRRC) after curative resection represents a difficult problem and a surgical challenge. The aim of this study was to evaluate the results of resecting the local recurrence of rectal cancer and to analyze factors that might predict curative resection and those that affect survival. PATIENTS AND METHODS: A retrospective review was performed in 50 patients who underwent surgical exploration with intent to cure LRRC between April 1998 and April 2005. All of the patients had previously undergone resection of primary rectal adenocarcinoma. Of these patients' charts, operation and pathology reports were reviewed. Primary tumor and treatment details, hospital of initial treatment and TNM stage were registered. The following data were collected concerning the detection of the local recurrence; date of recurrence, symptoms at the time of presentation and diagnostic work-up. Perioperative complication and date of discharge were also gathered. The recurrent tumors were classified as not fixed (F0), fixed at one site (F1) and fixed to two or more sites (F2) according to the preoperative and peroperative findings. Microscopic involvement of surgical margins and localization of recurrence were noted based on pathology reports. RESULTS: The median time interval between resection of primary tumor and surgery for locally recurrent disease was 24 (4-113) months. In a statistical analysis, initial surgery, complaints of patients, increasing number of sites of the recurrent tumor fixation in the pelvis, location of the recurrent tumor were associated with curative surgery. Curative, negative resection margins were obtained in 24 (48%) of patients; in these patients a median survival of 28 months was achieved, compared to 12 months (p=0.01) in patients with either microscopic or gross residual disease. Primary operation and CEA level at recurrence were also found to be important factors associated with improved survival. There was no operative mortality and, the complication rate was 24%. CONCLUSIONS: This study demonstrated that many patients with LRRC can be resected with negative margins. The type of primary surgery, symptoms, location, and fixity of recurrent tumor are associated with the increased possibility of carrying out curative resection. Previous surgery and curative surgery are significant predictors of both disease specific survival and overall survival. PMID- 17400424 TI - [On the difficulty of Traumatic brain injured patients end-of-life decisions]. AB - Traumatic brain injury occurs abruptly, involves multiple specialized teams, solicits the health care system in its emergency dimension and engages the well being of the patient and his relatives for a life time period. Clinicians are faced with issues of uppermost importance: medical issues such as predicting long term neurological outcome of the comatose patient, ethical issues because of the influence of intensive care on the long term survival of patients in vegetative and minimally conscious state, legal issues as the consequence of the current law which has set a new concept of proportionality of care, social issues as the result of the very high cost of these pathologies. This review will focus on the brain explorations that are required such as CT scan, evoked potentials, electroencephalography, magnetic resonance imaging and magnetic resonance spectroscopy to provide to the clinician a multimodal assessment of the brain state to predict outcome of coma. Such assessment is mandatory to answer the crucial question of proportionality of care in these patients. However, these techniques need further validation on large series of patients before being useful on clinical practice. PMID- 17400425 TI - Phonatory effects of supraglottic structures in excised canine larynges. AB - The aim of this study is to determine how phonation is affected by the presence and by alteration in the position of the supraglottic structures. The study used three excised canine larynges. A series of pressure-flow experiments were completed first on the excised larynx with false folds and epiglottis intact, then with the epiglottis removed, and finally with the false folds removed. Aerodynamic and acoustic effects were quantified with the analysis of the pressure, flow, and audio signals. The results of the study indicated that (1) elevation of the epiglottis to upright position from a horizontal position decreased subglottal pressure, increased flow (decreased laryngeal resistance), and slightly decreased fundamental frequency; (2) vibration of the false vocal folds induced some irregularity into the acoustic output of the larynx; (3) the presence of the epiglottis and the false vocal folds enhanced the second partial of the acoustic signal; and (4) the absence of the epiglottis and false folds increased low-frequency noise (between 0 and 300 Hz). Alteration in the position of the supraglottic structures affects laryngeal aerodynamics and acoustics, possibly due to biomechanical linkage with true vocal folds. When the supraglottic structures are present they act as resonators, enhancing the second partial and when they are absent (as in persons with supraglottic laryngectomy), low-frequency noise is increased perhaps due to the loss of boundary conditions or due to the presence of loose tissue. PMID- 17400426 TI - Evidence for distinguishing pressed, normal, resonant, and breathy voice qualities by laryngeal resistance and vocal efficiency in vocally trained subjects. AB - The purpose of this study was to determine if pressed, normal, resonant, and breathy voice qualities can be distinguished from one another by laryngeal resistance (LR; cm H(2)O/l/s) and/or vocal efficiency (VE; dB/cm H(2)O x l/s) in vocally, trained subjects. The experimental design was a within-subjects repeated measures design. Independent variables were pressed, normal, resonant, and breathy voice qualities. Dependent variables were LR and VE. Participants were 13 women of age 18-45 years with established vocal expertise. After a brief training phase, subjects were asked to produce each of the voice qualities on the pitch A3 (220 Hz) at a constant, individually identified comfortable dB level (+/-1 dB), during a repeated consonant-vowel utterance of /pi pi pi pi pi/. Results indicated that LR but not VE reliably distinguished pressed, normal, and breathy voice. Neither of the measures, however, distinguished normal from resonant voice, which were distinguished perceptually. The results suggest that LR may provide a useful tool for studying the coordinative dynamics of pressed, normal, and breathy voice qualities. PMID- 17400427 TI - Perception of voice in the transgender client. AB - Fundamental frequency (F(0)) of speech is used to measure the success of voice therapy in male-to-female transgender clients. This study evaluates the relationship between F(0) and patients' happiness with their voice. The study design used was a cross-sectional evaluation of client satisfaction questionnaires and voice recordings from transgender clients. This study was a comparative evaluation of voice recordings by voice professionals and lay observers. Twelve male-to-female transgender participants completed visual analogue scales (VASs), rating happiness with self-perceived femininity of their voice. Fifteen speech and language therapists (SLTs) and 40 naive observers evaluated the anonymized recordings, using the same rating system. The correlation between mean F(0) and participant happiness was established. Relationships between participant happiness and rater opinions were explored. A significant relationship between F(0) and participant happiness could not be demonstrated (r=0.32, P=0.32). There was a moderately strong positive correlation between self-perception of vocal femininity and perception of femininity by SLTs and naive observers (r=0.76 and 0.68, P=0.003 and P=0.01, respectively). This study demonstrates that happiness with voice in male-to-female transgender clients is not directly related to F(0). Clients can assess femininity of their voice in the form of perceived pitch. This may not affect happiness scores. Voice satisfaction may not correlate with perceptions of supervising voice professionals. However, professionals can reliably evaluate how the voice will be received by the lay public. Subjective measures of patient satisfaction, including VASs, are reliable and valid tools in evaluating therapeutic success. PMID- 17400428 TI - Rats bred for susceptibility to depression-like phenotypes have higher kainic acid-induced seizure mortality than their depression-resistant counterparts. AB - Epidemiological evidence suggests that epilepsy and depression are comorbid diseases. In fact, depression is the most common neuropsychiatric disorder associated with epilepsy, particularly temporal lobe epilepsy, and individuals with a history of depression are at a higher risk for developing epilepsy than the general population. Despite the epidemiological evidence for this link, there has been little experimental evidence to support the connection or elucidate possible underlying mechanisms. In an effort to address this problem and develop an animal model of epilepsy and depression comorbidity, we assessed seizure susceptibility and severity parameters in rats selectively bred for either susceptibility (the SwLo, SUS, and HYPER lines) or resistance (the SwHi, RES, and MON RES lines) to depression-like phenotypes. We found that rats bred for susceptibility to depression-like phenotypes experienced higher mortality following kainic acid-induced seizures than their resistant counterparts. In contrast, most line differences were not recapitulated when flurothyl was used to elicit seizures. Stress reduced kainic acid-induced mortality rates in all lines except the HYPER rats, supporting previously established indications that the stress response of HYPER rats is abnormal. These combined results support a neurobiological link between epilepsy and depression, advancing us towards an animal model of their comorbidity. PMID- 17400429 TI - Detection of decreased susceptibility to fluoroquinolones in Salmonella spp. by five different methods including real-time polymerase chain reaction (PCR). AB - Detection of Salmonella spp. isolates showing decreased susceptibility to fluoroquinolones has become important owing to the increasing prevalence of these strains and their association with treatment failure. Nalidixic acid agar dilution, nalidixic acid disk diffusion, MicroScan automated system and real-time polymerase chain reaction (PCR) (LightCycler) followed by melting temperature (Tm) analysis are compared with ciprofloxacin agar dilution as suitable methods to detect decreased susceptibility to fluoroquinolones in 100 Salmonella spp. isolates. Three minor discrepancies were found for nalidixic acid disk diffusion, one minor discrepancy was found for nalidixic acid agar dilution and Tm analysis, and one major discrepancy was found for MicroScan. Nalidixic acid disk diffusion was confirmed as a good screening method. Tm analysis is a rapid and accurate method for detecting decreased susceptibility to fluoroquinolones due to gyrA mutations in Salmonella spp. PMID- 17400430 TI - Point prevalence survey of antibiotic use in Scottish hospitals utilising the Glasgow Antimicrobial Audit Tool (GAAT). AB - A point prevalence survey of antimicrobial prescribing was performed in 10 Scottish hospitals using the Glasgow Antimicrobial Audit Tool (GAAT). Appropriateness of the intravenous (IV) route was determined by an infectious diseases physician (IDP) and by a computerised algorithm. The IDP also estimated IV agent appropriateness. Each hospital was surveyed on a single day. Of 3826 patients surveyed, 1079 (28.3%) received an antibiotic, 381 (35.3%) intravenously; 197 (28.2%) orally treated had prior IV therapy. Median duration of IV was 4 days (IQR 2-7 days) and oral switch was 3.5 days (2-6). IV route was appropriate in 84% (IDP) and 84.8% (algorithm). Choice of agent was appropriate in 80% (IDP). Third-generation cephalosporins (3GC) (28.3%) were most frequent, followed by co-amoxiclav (20.2%), metronidazole (19.2%) and glycopeptides (18.6%). Regional differences were seen. The study shows it is possible to coordinate, collect and compare data from UK hospitals using the GAAT. Data may usefully inform local and national audit and support prescribing initiatives. PMID- 17400431 TI - A photodimerization approach to crosslink and functionalize microgels. AB - Microgels were prepared within reverse micelles via photocrosslinking. Gelation resulted from the [2+2] photodimerization reaction of nitrocinnamoyl (NC) groups on multi-arm polyethylene glycol (PEG) or gelatin. Because of the potential for biomedical and chemical applications, immobilization capacity within the microgels was investigated. Quantum dots (QDs), for example, share a similar size scale with proteins and can be physically trapped within the microgels. In addition, the optoelectronic properties of QDs could be utilized for analytical, imaging, and therapeutic purposes. Small molecules and recognition sequences (e.g. biotin) can also be covalently immobilized within the microgel networks through the photodimerization reaction. In the presence of biotin-PEG-NC, the resulting microgels added to streptavidin-coated plates. The microgel properties such as biodegradability and degree of swelling may be engineered for particular applications including targeted monitoring and controlled drug delivery systems. PMID- 17400432 TI - Complex-valued wavelet artificial neural network for Doppler signals classifying. AB - OBJECTIVE: In this paper, the new complex-valued wavelet artificial neural network (CVWANN) was proposed for classifying Doppler signals recorded from patients and healthy volunteers. CVWANN was implemented on four different structures (CVWANN-1, -2, -3 and -4). MATERIALS AND METHODS: In this study, carotid arterial Doppler ultrasound signals were acquired from left carotid arteries of 38 patients and 40 healthy volunteers. The patient group had an established diagnosis of the early phase of atherosclerosis through coronary or aortofemoropopliteal angiographies. In implemented structures in this paper, Haar wavelet and Mexican hat wavelet functions were used as real and imaginary parts of activation function on different sequence in hidden layer nodes. CVWANN-1, -2 3 and -4 were implemented by using Haar-Haar, Mexican hat-Mexican hat, Haar Mexican hat, Mexican hat-Haar as real-imaginary parts of activation function in hidden layer nodes, respectively. RESULTS AND CONCLUSION: In contrast to CVWANN 2, which reached classification rates of 24.5%, CVWANN-1, -3 and -4 classified 40 healthy and 38 unhealthy subjects for both training and test phases with 100% correct classification rate using leave-one-out cross-validation. These networks have 100% sensitivity, 100% specifity and average detection rate is calculated as 100%. In addition, positive predictive value and negative predictive value were obtained as 100% for these networks. These results shown that CVWANN-1, -3 and -4 succeeded to classify Doppler signals. Moreover, training time and processing complexity were decreased considerable amount by using CVWANN-3. As conclusion, using of Mexican hat wavelet function in real and imaginary parts of hidden layer activation function (CVWANN-2) is not suitable for classifying healthy and unhealthy subjects with high accuracy rate. The cause of unsuitability (obtaining the poor results in CVWANN-2) is lack of harmony between type of activation function in hidden layer and type of input signals in neural network. PMID- 17400433 TI - Fractionated conformal radiotherapy in vestibular schwannoma: early results from a single centre. AB - AIMS: To assess the local control and cranial nerve toxicity in vestibular schwannoma patients treated with fractionated conformal radiotherapy delivered using a linear accelerator. MATERIALS AND METHODS: Ninety-five patients were referred for consultation to the Oncology Department in Addenbrookes Hospital between 1996 and 2005. The 42 cases who received fractionated conformal radiotherapy are the subject of this analysis. All patients had radiological or symptomatic progression. Conformal radiotherapy was prescribed at 50Gy in 30 fractions over 6 weeks, delivered using a linear accelerator. Patients were immobilised using either a beam direction shell or a Gill Thomas Cosman relocatable stereotactic head frame. RESULTS: The median age was 63 years (range 28-81) with 57% men. The average tumour size was 21.5mm on magnetic resonance imaging. Before treatment, 20 (48%) patients were deemed to have useful hearing on the affected side. The median follow-up was 18.6 months (range 0.3-6.5 years) and the actuarial local control rate at 2.5 years was 96.9% (one patient progressed after treatment). In previously hearing patients, the actuarial rate of useful hearing preservation was 100%, and the rate of mild hearing loss was 20% at 1 year and 26.7% at 2.5 years of follow-up. There were five neurofibromatosis type 2 patients treated, two of whom had useful hearing before radiotherapy. In one patient this was affected, with a 20dB loss, although he still has useful hearing. In those with normal facial nerve function before radiotherapy (n=40), this was preserved in 96.8% at 2.5 years. Trigeminal nerve function was preserved in all patients (n=38) who had normal nerve function before radiotherapy. CONCLUSION: Although follow-up was relatively short in this single institution series, fractionated linear accelerator radiotherapy gave excellent local control, useful hearing preservation and retained cranial nerve function in vestibular schwannoma. PMID- 17400434 TI - sP-selectin plasma levels in obesity: association with insulin resistance and related metabolic and prothrombotic factors. AB - BACKGROUND AND AIM: Soluble P-selectin (sP-sel) represents a marker of platelet activation. This study was addressed to investigate the associations of sP-sel plasma levels with anthropometric parameters, insulin resistance, and related metabolic and prothrombotic factors. METHODS AND RESULTS: 50 non-diabetic women, 17 with normal weight and 33 overweight and obese, aged 18-55 years, were examined. Measurements included body mass index (BMI), central fat accumulation (evaluated by waist circumference), systolic and diastolic blood pressure levels, fasting plasma concentrations of sP-sel, glucose, lipids (triglycerides, total cholesterol and HDL-cholesterol), insulin, and prothrombotic factors (plasminogen activator inhibitor-1, von Willebrand factor, fibrinogen), and insulin resistance (estimated by the homeostasis model assessment: HOMA(IR)). Overweight and obese women had higher fasting plasma sP-sel concentrations compared to normal-weight controls (P<0.05). sP-sel concentrations were positively correlated with BMI, HOMA(IR), systolic and diastolic blood pressure, fasting insulin, triglyceride and PAI-1 plasma levels (P<0.05 for all the correlations). When a multiple regression analysis was performed, with P-sel as dependent variable and all the other parameters as independent variables, P-sel did not maintain a significant relationship with any of these variables. CONCLUSIONS: s-P-selectin plasma concentrations are higher in overweight and obese insulin resistant subjects, thus possibly contributing to the cardiovascular risk of these patients. However, body fatness and insulin resistance are not independent determinants of fasting plasma sP-sel concentrations. PMID- 17400435 TI - Mechanical properties of the endophytic ovipositor in damselflies (Zygoptera, Odonata) and their oviposition substrates. AB - Damselfly females use their ovipositor valves to saw aquatic plants in order to insert their eggs into the plant tissues. Stiffness of the plant substrata is therefore an important parameter for oviposition substrate choice by females. Using a force transducer combined with a motorised micromanipulator, the bending stiffness of the ovipositor at the axial compressional load was studied in seven European damselfly species and compared to the local stiffness of seven preferred plant substrates. The puncture force of tested plant samples ranged from 105 to 1500 mN, and their local stiffness ranged from 208 to 1776 N/m. The bending stiffness of the ovipositor was estimated as 173-409 N/m depending on the damselfly species. Using original and literature data, a significant positive correlation between mechanical properties of the ovipositor and preferred oviposition substrates was demonstrated. Possible behavioural adaptations to overcome high stiffness of plant tissues during oviposition are discussed. PMID- 17400436 TI - Head capsule, chephalic central nervous system and head circulatory system of an aberrant orthopteran, Prosarthria teretrirostris (Caelifera, Hexapoda). AB - The head capsule, the circulatory system and the central nervous system of the head of Prosarthria teretrirostris (Proscopiidae) is described in detail, with special consideration of modifications resulting from the aberrant head shape. The transformations of the head are completely different from those found in phasmatodeans, which are also characterised by twig mimesis. The circulatory system is distinctly modified. A hitherto undescribed additional structure in the posterior head region very likely functions as a pulsatile organ. The cephalic central nervous system is strongly elongated, with changes in the position of the suboesophageal ganglion, the corpora cardiaca and the course of the nervus mandibularis. Three-dimensional reconstructions of these two organ systems in combination with the pharynx were made using Alias Maya 6.0 software. Comparisons with other representatives of Caelifera suggest a clade comprising Proscopiidae and Morabinae. The presence of a transverse muscle connecting the antennal ampullae in Prosarthria shows that this structure likely belongs to the groundplan of Orthoptera, even though it is missing in different representatives of this group. The transverse ampullary muscle is a potential synapomorphy of Orthoptera, Phasmatodea and Dictyoptera. PMID- 17400437 TI - Effects of constructed wetland system on the removal of dibutyl phthalate (DBP). AB - Phthalic acid esters (PAEs) have become widely diffused in the environment via the manufacturing process. Numerous experiments have shown that the bioaccumulation of PAEs occurred in the aquatic and terrestrial food chain; meanwhile, it was found that some of PAEs were considered as potential carcinogens, teratogens and mutagens. In this research, two vertical/reverse vertical flow constructed wetland systems were set up to study its removal efficiency of dibutyl phthalate (DBP) pollution. The results showed that the constructed wetland system could remove DBP effectively, and the removal rates reached nearly 100%. Substrate microorganism and enzymatic activities probably played key roles during DBP removal, and the removal of DBP probably mainly took place in the upper layer of chamber A in the constructed wetland systems. PMID- 17400438 TI - Molecular characterization of the BvgA response regulator of Bordetella holmesii. AB - The BvgAS system controls the expression of most virulence factors in Bordetella pertussis. Recently, we identified an orthologous system in the related human pathogen Bordetella holmesii. However, while we found that the orthologous histidine kinases BvgS could be functionally exchanged between the two species, the B. holmesii response regulator BvgA(BH) could not substitute for its B. pertussis counterpart in vivo and, accordingly, was not able to bind to B. pertussis virulence promoters in vitro. Here we show that a hybrid response regulator consisting of the B. pertussis derived DNA-binding output domain of BvgA(BP) combined with the B. holmesii receiver domain binds to BvgA(BP) regulated virulence promoters of B. pertussis in vitro and is functional in B. pertussis in vivo. This shows that the inability of BvgA(BH) to complement BvgA(BP) in B. pertussis is due to the small number of sequence variations present in its output domain. However, by mutation analysis we show that four amino acid exchanges present in the helix-turn-helix motif of BvgA(BH) as compared to BvgA(BP) are not the only reason for its inability to substitute for BvgA(BP) but additional mutations present in the output domain must play a role. PMID- 17400439 TI - Patterning of proteins and cells on functionalized surfaces prepared by polyelectrolyte multilayers and micromolding in capillaries. AB - A method for protein and cell patterning on polyelectrolyte-coated surfaces using simple micromolding in capillaries (MIMIC) is described. MIMIC produced two distinctive regions. One contained polyethylene glycol (PEG) microstructures fabricated using photopolymerization that provided physical, chemical, and biological barriers to the nonspecific binding of proteins, bacteria, and fibroblast cells. The second region was the polyelectrolyte (PEL) coated surface that promoted protein and cell immobilization. The difference in surface functionality between the PEL region and background PEG microstructures resulted in simple patterning of biomolecules. Fluorescein isothiocyanate-tagged bovine serum albumin, E. coli expressing green fluorescence protein (GFP), and fibroblast cells were successfully bound to the exposed PEL surfaces at micron scale. Compared with the simple adsorption of protein, fluorescence intensity was dramatically improved (by about six-fold) on the PEL-modified surfaces. Although animal cell patterning is prerequisite for adhesive protein layer to survive on desired area, the PEL surface without adhesive proteins provides affordable microenvironment for cells. The simple preparation of functionalized surface but universal platform can be applied to various biomolecules such as proteins, bacteria, and cells. PMID- 17400440 TI - The region ion sensitive field effect transistor, a novel bioelectronic nanosensor. AB - A novel type of bioelectronic region ion sensitive field effect transistor (RISFET) nanosensor was constructed and demonstrated on two different sensor chips that could measure glucose with good linearity in the range of 0-0.6mM and 0-0.3mM with a limit of detection of 0.1 and 0.04 mM, respectively. The sensor is based on the principle of focusing charged reaction products with an electrical field in a region between the sensing electrodes. For glucose measurements, negatively charged gluconate ions were gathered between the sensing electrodes. The signal current response was measured using a low-noise pico ammeter (pA). Two different sizes of the RISFET sensor chips were constructed using conventional electron beam lithography. The measurements are done in partial volumes mainly restricted by the working distance between the sensing electrodes (790 and 2500 nm, respectively) and the influence of electrical fields that are concentrating the ions. The sensitivity was 28 pA/mM (2500 nm) and 830 pA/mM (790 nm), respectively. That is an increase in field strength by five times between the sensing electrodes increased the sensitivity by 30 times. The volumes expressed in this way are in low or sub femtoliter range. Preliminary studies revealed that with suitable modification and control of parameters such as the electric control signals and the chip electrode dimensions this sensor could also be used as a nanobiosensor by applying single enzyme molecule trapping. Hypotheses are given for impedance factors of the RISFET conducting channel. PMID- 17400441 TI - Ventricular bigeminy during phenylephrine infusion used to maintain normotension during caesarean section under spinal anaesthesia. AB - A 31-year-old primiparous, healthy woman presented for emergency caesarean section. Following the siting of a spinal anaesthetic, seconds after starting a phenylephrine infusion, she developed ventricular bigeminy. She reverted to sinus rhythm spontaneously when the phenylephrine infusion was stopped at delivery. The possible proarrhythmic and antiarrhythmic effects of phenylephrine are discussed. We suggest that this was most probably a stretch-induced ventricular arrhythmia due to increased ventricular afterload. PMID- 17400442 TI - Obesity and prostate cancer: an ambiguous relationship. PMID- 17400443 TI - Inflammation and cancer: is trophism the link? AB - The pathophysiological mechanisms of the inflammatory response can be common to wound repair and tumor development. We propose that this response evolves in three phases, the nervous or immediate phase, the immune or intermediate phase, and the endocrine or late phase. In wound repair and in these phases, the interstitial space successively presents edema due to ischemia-revascularization and nutrition by diffusion (nervous phase), infiltration by leukocytes, which would mediate the nutrition of damaged neighbor cells (immune phase) and by angiogenesis, nutrition mediated by the capillaries that favor regeneration or scarring (endocrine phase). At the same time, in tumor development, it is considered that the cancerous cell successively occupies the interstitial space, expressing three different phenotypes: the hypoxia-reperfusion phenotype, with anaerobic glycolisis, oxidative stress and edema (dormant stage); the immune phenotype that expresses the functions corresponding to leukocytes, including the hyperproduction of pro-inflammatory mediators, lymphangiogenesis, the invasion of lymph nodes (N stage) and systemic inflammatory response syndrome; and lastly, the endocrine phenotype, in which the appearance of both local (tumor or T stage) and systemic (metastasis or M stage) angiogenesis induce a growing disease. PMID- 17400444 TI - Reduction of heavy metal contents in liquid effluents by vermicomposts and the use of the metal-enriched vermicomposts in lettuce cultivation. AB - The removal of Cu, Ni and Zn from electroplating effluents by adsorption in cattle manure vermicompost has been discussed. A glass column 38 cm long and 7 cm i.d. was loaded with cattle manure vermicompost and effluents were passed through it. The metal concentrations were measured in the elutant. The experiments on adding effluent aliquots into the columns were continued until the metal concentrations in the elutant reached the maximum values established for effluent discharges in water courses by the Brazilian quality criteria, i.e., Cu=1.0 mg L( 1), Ni=2.0 mg L(-1), and Zn=5.0 mg L(-1). The amount of Cu retention by the vermicompost was determined at the natural effluent pH (2.0). The Zn and Ni retentions were evaluated at the natural effluent pH (6.9 and 7.4, respectively) as well pH 2.0. Vermicompost residues obtained from this process were used for lettuce cultivation. The vermicompost was found to be efficient in removing metals from the electroplating wastes, as well as in the increase of its pH values. Metal retention values were close to 100%. The Cu concentrations in lettuce leaves from the treatment with vermicompost enriched with this metal were below the range of critical toxicity level to plants, i.e., from 20 to 100 mg L( 1). However, the estimated Cu concentrations in the roots from the treatment with vermicompost enriched with Cu were much larger than that of the treatment with the natural vermicompost, reaching 246.3 mg L(-1). The Ni and Zn concentrations in lettuce leaves from the treatments, with vermicomposts enriched with the respective metals, were above the range of critical toxicity levels to plants, i.e., from 10 to 50 mg kg(-1) and from 15 to 30 mg kg(-1), respectively. However, no symptom of toxicity was found visually. Larger accumulations of Cu, Ni and Zn were found in the lettuce leaves than in the roots after the treatments with the uncontaminated vermicompost. A greater absorption of Cu and Ni by roots was found in treatments with vermicompost enriched with these elements, whereas Zn was found preferentially in the leaves. The statistical analysis was done by analyses of variance and regression. PMID- 17400445 TI - The fate of crop nutrients during digestion of swine manure in psychrophilic anaerobic sequencing batch reactors. AB - The objectives of the study were to measure the levels of manure nutrients retained in psychrophilic anaerobic sequencing batch reactors (PASBRs) digesting swine manure, and to determine the distribution of nutrients in the sludge and supernatant zones of settled bioreactor effluent. Anaerobic digestion reduced the total solids (TS) concentration and the soluble chemical oxygen demand (SCOD) of manure by 71.4% and 79.9%, respectively. The nitrogen, potassium, and sodium fed with the manure to the PASBRs were recovered in the effluent. The bioreactors retained on average 25.5% of the P, 8.7% of the Ca, 41.5% of the Cu, 18.4% of the Zn, and 67.7% of the S fed to the PASBRs. The natural settling of bioreactor effluent allowed further nutrient separation. The supernatant fraction, which represented 71.4% of effluent volume, contained 61.8% of the total N, 67.1% of the NH4-N, and 73.3% of the Na. The settled sludge fraction, which represented 28.6% of the volume, contained 57.6% of the solids, 62.3% of the P, 71.6% of the Ca, 89.6% of the Mg, 76.1% of the Al, 90.0% of the Cu, 74.2% of the Zn, and 52.2% of the S. The N/P ratio was increased from 3.9 in the raw manure to 5.2 in the bioreactor effluent and 9.2 in the supernatant fraction of the settled effluent. The PASBR technology will then substantially decrease the manure management costs of swine operations producing excess phosphorus, by reducing the volume of manure to export outside the farm. The separation of nutrients will also allow land spreading strategies that increase the agronomic value of manure by matching more closely the crop nutrient requirements. PMID- 17400446 TI - Removal of basic and reactive dyes using ethylenediamine modified rice hull. AB - Wastewaters from textile industries may contain a variety of dyes that have to be removed before their discharge into waterways. Rice hull, an agricultural by product, was modified using ethylenediamine to introduce active sites on its surface to enable it to function as a sorbent for both basic and reactive dyes. The sorption characteristics of Basic Blue 3 (BB3) and Reactive Orange 16 (RO16) by ethylenediamine modified rice hull (MRH) were studied under various experimental conditions. Sorption was pH and concentration dependent. Simultaneous removal of BB3 and RO16 occurred at pH greater than 4. The kinetics of dye sorption fitted a pseudo-second order rate expression. Increase in agitation rate had no effect on the sorption of BB3 but increased uptake of RO16 on MRH. Decreasing particle size increased the uptake of dyes in binary dye solutions. Equilibrium data could be fitted into both the Langmuir and Freundlich isotherms. Maximum sorption capacities calculated from the Langmuir model are 14.68 and 60.24 mg/g for BB3 and RO16, respectively in binary dye solutions. This corresponds to an enhancement of 4.5 and 2.4 fold, respectively, compared to single dye solutions. MRH therefore has the potential of being used as an efficient sorbent for the removal of both dyes in textile wastewaters. PMID- 17400447 TI - Vermicompost as a soil supplement to improve growth, yield and fruit quality of tomato (Lycopersicum esculentum). AB - The effects of earthworm-processed sheep-manure (vermicompost) on the growth, productivity and chemical characteristics of tomatoes (Lycopersicum esculentum) (c.v. Rio Grande) were investigated in a greenhouse experiment. Five treatments were applied combining vermicompost and soil in proportions of 0:1, 1:1, 1:2, 1:3, 1:4 and 1:5 (v/v). Growth and yield parameters were measured 85 days and 100 days after transplanting. Addition of vermicompost increased plant heights significantly, but had no significant effect on the numbers of leaves or yields 85 days after transplanting. Yields of tomatoes were significantly greater when the relationship vermicompost:soil was 1:1, 1:2 or 1:3, 100 days after transplanting. Addition of sheep-manure vermicompost decreased soil pH, titratable acidity and increased soluble and insoluble solids, in tomato fruits compared to those harvested from plants cultivated in unamended soil. Sheep manure vermicompost as a soil supplement increased tomato yields and soluble, insoluble solids and carbohydrate concentrations. PMID- 17400448 TI - Removal characteristics of metal ions using degreased coffee beans: adsorption equilibrium of cadmium(II). AB - The feasibility of using coffee beans after being dripped and degreased (DCB) as an adsorbent for base metals such as copper(II), zinc(II), lead(II), iron(III) and cadmium(II) were examined. The compositions of the DCB were characterized by Fourier transform infrared spectroscopy, scanning electronic micrograph and fluorescent X-ray. It was found that DCB contain sulfur and calcium from the analysis using fluorescent X-ray. The plant cell wall in DCB has the porous structure from the scanning electron microscopy (SEM) analysis, and the specific surface area was determined to be 1.2 m2/g using the specific surface area analyzer. Batch adsorption experiments on DCB were carried out at various pHs in order to elucidate the selectivity of metal ions. All metals were adsorbed at low pH region (3.0-5.0). Of particular interest was the adsorption characteristics of cadmium(II) on DCB. The adsorption isotherm for cadmium(II) at pH 8 fitted with a Langmuir equation to yield an adsorption equilibrium constant of 55.2 mmol dm(-3) and an adsorption capacity of 5.98 x 10(-2) mmol g(-1). The desorption of cadmium(II) was easily achieved over 90% by a single batchwise treatment with an aqueous solution of hydrochloric acid or nitric acid at more than 0.01 mol dm( 3). These results suggested that DCB behaves as a cation exchanger. PMID- 17400449 TI - Biotransformation of gentiopicroside by asexual mycelia of Cordyceps sinensis. AB - The biotransformation of gentiopicroside by asexual mycelia of Cordyceps sinensis yielded two products, one of which was proved to be a new pyridine monoterpene alkaloid. The possible mechanisms were discussed. PMID- 17400450 TI - Preparation of novel anthranilic acids as antibacterial agents: extensive evaluation of structural and physical properties on antibacterial activity and human serum albumin affinity. AB - In the past few years a significant effort has been devoted by Pharmacia toward the discovery of novel antibiotics. We have recently described the identification of an anthranilic acid lead 1 and the optimization resulting in the advanced lead 2. In this report, we describe the preparation of several selected analogs to probe the dependency of this template for antibacterial activity and the affinity these compounds have for human serum albumin (HSA). These analogs illustrate that decreased affinity for HSA can be achieved while retaining relevant antibacterial activity. The most important factor for reduced HSA affinity is decrease in logP rather than a structural change. PMID- 17400453 TI - Additional interaction of allophenylnorstatine-containing tripeptidomimetics with malarial aspartic protease plasmepsin II. AB - Based on a highly potent allophenylnorstatine-containing inhibitor, KNI-10006, against the plasmepsins of Plasmodium falciparum, we synthesized a series of tripeptide-type compounds with various N-terminal moieties and evaluated their inhibitory activities against plasmepsin II. Certain phenylacetyl derivatives exhibited extremely high affinity with K(i) values of less than 0.1n M suggesting successful hydrophobic interactions. PMID- 17400451 TI - Potent anti-obese principle from Rosa canina: structural requirements and mode of action of trans-tiliroside. AB - The 80% aqueous acetone extracts from the fruit (50 mg/kg/d) and seeds (12.5 and 25 mg/kg/d) of Rosa canina L., but not from the pericarps, were found to show substantial inhibitory effect on the gain of body weight and/or weight of visceral fat without affecting food intake in mice for 2 weeks after administration of the extracts. With regard to the active constituents, the principal constituent, trans-tiliroside (0.1-10 mg/kg/d), potently inhibited the gain of body weight, especially visceral fat weight, and significantly reduced blood glucose levels after glucose loading (1 g/kg, ip) in mice. On the other hand, kaempferol and p-coumaric acid lacked such effect and kaempferol 3-O-beta-D glucopyranoside tended to reduce the gain of body weight and visceral fat weight, but not significantly, at a dose of 10 mg/kg/d. These results indicate the importance of both kaempferol 3-O-beta-D-glucopyranoside and p-coumaroyl moieties for anti-obese effects. Furthermore, a single oral administration of trans tiliroside at a dose of 10 mg/kg increased the expression of PPAR-alpha mRNA of liver tissue in mice. PMID- 17400452 TI - PDE-10A inhibitors as insulin secretagogues. AB - Modulation of cAMP levels has been linked to insulin secretion in preclinical animal models and in humans. The high expression of PDE-10A in pancreatic islets suggested that inhibition of this enzyme may provide the necessary modulation to elicit increased insulin secretion. Using an HTS approach, we have identified quinoline-based PDE-10A inhibitors as insulin secretagogues in vitro. Optimized compounds were evaluated in vivo where improvements in glucose tolerance and increases in insulin secretion were measured. PMID- 17400454 TI - 6-hydroxy to 6'''-amino tethered ring-to-ring macrocyclic aminoglycosides as probes for APH(3')-IIIa kinase. AB - Based on molecular modeling and available X-ray structure data on aminoglycosides complexed with a bacterial ribosomal surrogate or with a kinase, two analogues of paromomycin were prepared by tethering the 6-OH and the 6'''-NH(2) group with a five-carbon bridge. Only one of two possible hydroxyl groups was phosphorylated by the kinase. The application of ring closure metathesis is presented for the first time to construct bridged macrocyclic analogues in the aminoglycoside series. PMID- 17400455 TI - Cobaltpolyoxometalate-catalyzed cyclization of glucal with aryl amines: Synthesis of 2,4-disubstituted tetrahydroquinolines. AB - Treatment of 3,4,6-tri-O-acetyl-d-glucal (1) with various aryl amines in the presence of dodecatungstocobaltate under mild and neutral conditions gave sugar derived tetrahydroquinoline derivates. PMID- 17400456 TI - Changing Australian prescribing patterns for antiepileptic drugs in pregnancy and their possible consequences. AB - We report progress in the accumulation of data by the Australian Pregnancy Register over 64 months, confirming the rise in enrollment and the predominantly epileptic indication for taking antiepileptic drugs. Eighty percent of the enrollment was prospective. The focus of the current report is the observation that as a possible result of education and dissemination of information about the risks of exposure to high-dose valproate, there has been a decline in the drug's doses prescribed in Australia, as well as a decline in the proportion of patients prescribed this drug in pregnancy. The risk of teratogenicity associated with valproate in doses in excess of 1100 mg/day was confirmed, and the incidence of lamotrigine-related malformations was comparable to that associated with exposure to phenytoin and carbamazepine. Reporting of data for this paper took into account the 12 months follow-up period for each pregnancy outcome, thus in effect making the evaluation period 21 months for each pregnancy and its outcome. PMID- 17400457 TI - Synthesis of isoquinuclidine analogs of chloroquine: antimalarial and antileishmanial activity. AB - The isoquinuclidine (2-azabicyclo[2.2.2]octane) ring system may be viewed as a semi-rigid boat form of the piperidine ring and, when properly substituted, a scaffold for rigid analogs of biologically active ethanolamines and propanolamines. It is present in natural products (such as ibogaine and dioscorine) that display interesting pharmacological properties. In this study, we have expanded our continuing efforts to incorporate this ring system in numerous pharmacophores, by designing and synthesizing semirigid analogs of the antimalarial drug chloroquine. The analogs were tested in vitro against Plasmodium falciparum strains and Leishmania donovani promastigote cultures. Compounds 6 and 13 displayed potent antimalarial activity against both chloroquine-susceptible D6 and the -resistant W2 strains of P. falciparum. All analogs also demonstrated significant antileishmanial activity with compounds 6 and 13 again being the most potent. The fact that these compounds are active against both chloroquine-resistant and chloroquine-sensitive strains as well as leishmanial cells makes them promising candidates for drug development. PMID- 17400458 TI - Polyphenols based on isoflavones as inhibitors of Helicobacter pylori urease. AB - Twenty polyphenols were synthesized and evaluated for their effect on Helicobacter pylori urease. Among these compounds, 4-(p hydroxyphenethyl)pyrogallol (15) (IC(50)=0.03 mM) and 7,8,4'-trihydroxyisoflavone (19) (IC(50)=0.14 mM) showed potent inhibitory activities, and inhibited Helicobacter pylori urease in a time-dependent manner. The structure-activity relationship of these polyphenols revealed: the two ortho hydroxyl groups were essential for inhibitory activity of polyphenol. When the C-ring of isoflavone was broken, the inhibitory activity markedly decreased. As for deoxybenzoin, the carboxyl group was clearly detrimental. PMID- 17400459 TI - Synthesis and antimicrobial activity of symmetrical two-tailed dendritic tricarboxylato amphiphiles. AB - Two series of water-soluble, symmetrical two-tailed homologous dendritic amphiphiles--R(2)NCONHC((CH(2))(2)COOH)(3), 2(n,n), and R(2)CHNHCONHC((CH(2))(2)COOH)(3), 3(n,n), where R=n-C(n)H(2n+1)--were synthesized and compared to R''NHCONHC((CH(2))(2)COOH)(3), 1(n), R''=n-C(n)H(2n+1), to determine whether antimicrobial activity was influenced by total or individual alkyl chain lengths, and whether antimicrobial activity depends on hydrophobicity or tail topology (one or two). In a broad screen of 11 microorganisms, 2(n,n) and 3(n,n) generally displayed higher minimal inhibitory concentrations (MICs) than 1(n) against growth as measured by broth microdilution assays. Chain-length specificity was observed against Candida albicans as 1(16), 2(8,8), and 3(8,8) showed the lowest MIC in their respective series. The one case where two-tailed compounds displayed the lowest MICs-3(10,10), 15 microM; 3(11,11), 7.2microM; and 3(12,12), 6.9 microM-was against Cryptococcus neoformans. PMID- 17400460 TI - Radical scavenging and cytochrome P450 3A4 inhibitory activity of bergaptol and geranylcoumarin from grapefruit. AB - Grapefruit juice has been shown to increase the oral bioavailability of several clinically important drugs by inhibiting first pass metabolism. Several compounds in grapefruit juice have shown different biological activities. Unique among them are furocoumarins with potent inhibitory activity against cytochrome P450 enzymes. In the present study, two bioactive compounds were isolated from grapefruit juice and grapefruit peel oil. The purity of the isolated compounds has been analyzed by HPLC. Structures of the compounds were elucidated by extensive NMR and mass spectral studies and identified as bergaptol and geranylcoumarin. The isolated compounds were tested for their radical scavenging activity using 2,2'-azobis (3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazil (DPPH) methods at different concentrations. Bergaptol showed very good radical scavenging activity at all the tested concentrations. Furthermore, these compounds were evaluated for their inhibitory activity against CYP3A4 enzyme. Bergaptol and geranylcoumarin were found to be potent inhibitors of debenzylation activity of CYP3A4 enzyme with an IC(50) value of 24.92 and 42.93 microM, respectively. PMID- 17400461 TI - 9-Nitroanthracene derivative as a precursor of anthraquinone for photodynamic therapy. AB - Anthraquinones are typical photosensitizers used in photodynamic therapy (PDT). However, systemic toxicity is a major problem for anthraquinones due to their ability not only to bind DNA but also to cause oxidative stress even without photoirradiation. To avoid such disadvantages in cancer therapy, we designed and synthesized a novel 9-nitroanthracene derivative (1) as a precursor of anthraquinone. Under photoirradiation, 1 is converted into anthraquinone via generation of nitric oxide as confirmed by ESR. Strong DNA cleavage specifically at guanine under photoirradiation was also observed, characteristic of DNA cleaving reactions by photoirradiated anthraquinones. We propose development of 1 as an alternative approach toward PDT that reduces the systemic toxicity of anthraquinone. PMID- 17400462 TI - The isolation of secondary metabolites and in vitro potent anti-cancer activity of clerodermic acid from Enicosanthum membranifolium. AB - Four compounds were isolated from Enicosanthum membranifolium. The structures of the compounds were confirmed by spectroscopic data. Their structures were determined as N-trans-feruloyltyramine, R-(-)-mellein, clerodermic acid, and salicifoline chloride as a quaternary alkaloid compound. The structures of R-(-) mellein and salicifoline chloride were confirmed by using X-ray diffraction. Clerodermic acid was shown to induce potent apoptosis against human leukemia HL60 cells. PMID- 17400463 TI - Synthesis and antiproliferative activity of some novel derivatives of diospyrin, a plant-derived naphthoquinonoid. AB - Derivatisation of diospyrin, a bisnaphthoquinonoid isolated from Diospyros montana Roxb., led to the modification of its inhibitory activity, in vitro, towards a murine tumour model, Ehrlich ascites carcinoma (EAC), and two human cancer cell lines, viz., malignant skin melanoma (A375) and epidermoid laryngeal carcinoma (Hep2). Among the novel derivatives, an epoxide exhibited the maximum antiproliferative activity (IC(50) values in the range of 0.03-0.21 microM) and a comparatively lower toxicity (IC(50) approximately 98 microM) in normal human peripheral blood mononuclear cells (PBMC). This compound might provide a novel 'lead' for the development of clinically effective antiproliferative agents against cancer. PMID- 17400464 TI - A trypsin and chymotrypsin inhibitor from Caesalpinia bonduc seeds: isolation, partial characterization and insecticidal properties. AB - Evolution of proteinase inhibitor diversity in leguminous plants of tropical rainforests is under immense pressure from the regular upregulation of proteolytic machinery of their pests. The present study illustrates the isolation and bioinsecticidal potency of a serine proteinase inhibitor from the seeds of Caesalpinia bonduc (CbTI), inhabiting Great Nicobar Island, India. Following initial fractionation by ammonium sulfate precipitation, CbTI was purified to homogeneity by ion exchange, gel filtration and trypsin affinity chromatography. SDS-PAGE of gel filtrated CbTI showed a couple of proteins CbTI-1 ( approximately 16kDa) and CbTI-2 (20kDa) under non-reducing conditions, which subsequent to trypsin affinity chromatography yielded only CbTI-2. Both Native PAGE as well as iso-electric focusing showed 2 iso-inhibitors of CbTI-2 (pI values of 5.35 and 4.6). CbTI exhibited tolerance to extremes of temperatures (0-60 degrees C) and pH (1-12). A 1:1 stoichiometric ratio was noted during CbTI-2-trypsin complex formation, which was absent on binding with chymotrypsin. Further, SDS-PAGE analysis also showed that CbTI-1 has affinity only towards chymotrypsin, whereas both trypsin and chymotrypsin formed complexes with CbTI-2. Dixon plot analysis of CbTI-2 yielded inhibition constants (K(i)) of 2.75 x 10(-10)M and 0.95 x 10( 10)M against trypsin and chymotrypsin activity respectively. Preliminary investigations on the toxicological nature of CbTI revealed it to be a promising bioinsecticidal candidate. PMID- 17400465 TI - Evaluation of somatic hybrids of potato with Solanum stenotomum after a long-term in vitro conservation. AB - Somatic hybrids of potato with a cultivated relative, Solanum stenotomum also called Solanum tuberosum Stenotomum group, were evaluated for their physiological and agronomical characteristics as well as the stability of the introgressed resistance to bacterial wilt, caused by Ralstonia solanacearum, after a long-term in vitro conservation for more than 5 years. Analysis of photosynthesis showed that the PEPC/Rubisco ratio remained lower than 0.5 for all vitroplants of potato and the somatic hybrids, except for the relative species. This indicates that the carbon metabolism is heterotrophic (ratio>1) for S. stenotomum, and autotrophic for potato and the somatic hybrids (ratio<1). In both in vitro and greenhouse conditions, potato and the somatic hybrids produced few bigger tubers, while many small tubers were obtained from the relative. The hybrid tubers were morphologically intermediate. The starch content of hybrid tubers was much lower than that of potato, but similar to that of the relative species. Interestingly, the level of bacterial resistance, introgressed from S. stenotomum into potato, was shown to be very stable and remained as high as that of the relative after a long-term period of in vitro conservation. PMID- 17400466 TI - Isolated Medicago truncatula mutants with increased calcium oxalate crystal accumulation have decreased ascorbic acid levels. AB - The mechanisms controlling oxalate biosynthesis and calcium oxalate formation in plants remain largely unknown. As an initial step toward gaining insight into these regulatory mechanisms we initiated a mutant screen to identify plants that over-accumulate crystals of calcium oxalate. Four new mutants were identified, from an ethyl methanesulfonate (EMS)-mutagenized Medicago truncatula (cv. Jemalong genotype A17) population, that over-accumulated calcium oxalate crystals. The increased calcium oxalate content of these new mutants, as with the previously isolated mutant cod4, resulted from an increase in druse crystals accumulated within the mesophyll cells of leaves. Complementation and segregation analysis revealed that each mutant was affected at a different locus. This was confirmed through the genetic mapping of each mutation to different linkage groups. Together, these findings emphasize the complexity of factors that can contribute to oxalate biosynthesis and crystal formation in these plants. In addition, each mutant showed a common decrease in ascorbic acid content providing genetic support for ascorbic acid as a precursor in the oxalate biosynthetic pathway for druse crystal formation. Further support was obtained by the ability of an exogenous supply of ascorbate to induce druse crystal formation while other tested organic acids did not induce crystal production. PMID- 17400467 TI - Cellular ascorbic acid regulates the activity of major peroxidases in the apical poles of germinating white spruce (Picea glauca) somatic embryos. AB - In previous studies we have reported that applications of ascorbic acid (ACS) enhance the conversion frequency of white spruce somatic embryos by "rescuing" structurally disorganized meristems and inducing cell proliferation in the apical poles [C. Stasolla, E.C. Yeung, Ascorbic acid improves the conversion of white spruce somatic embryos, In Vitro Cell. Dev. Biol. Plant 35 (1999) 316-319]. In order to determine if the role played by this metabolite during embryo conversion is mediated by cellular peroxidases, the activity of guaiacol-, ferulic acid-, and ascorbic acid-dependent peroxidases were measured in the apical poles of germinating embryos with altered ASC levels. Changes in the endogenous ASC pool were achieved by treating the embryos with exogenously supplied ASC, L-galactono gamma-lactone (GL) the last precursor of the de novo biosynthesis of ASC, and lycorine (L), an inhibitor of the last reaction leading to the synthesis of ASC. Our studies demonstrate the existence of a negative correlation between cellular ASC levels and activities of both guaiacol and ferulic acid peroxidases in root and shoot apices. A depletion of cellular ASC enhanced the rate of both guaiacol and ferulic acid oxidation at the apical poles of the embryos and resulted in meristem abortion. In contrast, the activity of guaiacol and ferulic acid peroxidases decreased below control levels if the endogenous ASC content of the embryos was experimentally increased. Fluctuations of total peroxidase activity following alterations in ASC pool were also confirmed by histochemical staining and in vitro studies. Overall our results suggest that a threshold of ASC level must be maintained in the apical poles of germinating embryos in order to inhibit peroxidase activities from cross-linking cell wall components and preventing post embryonic growth. PMID- 17400468 TI - Preliminary experience with inhaled milrinone in cardiac surgery. AB - BACKGROUND: Inhaled administration of milrinone reduces pulmonary artery pressure. Pulmonary hypertension (PH) and right heart failure are associated with difficult separation from cardiopulmonary bypass (CPB). Therefore, inhaled milrinone could facilitate separation from CPB. OBJECTIVE: To determine the impact and timing of administration of inhaled milrinone. METHODS: A retrospective analysis of our experience on high-risk patients receiving inhaled milrinone was conducted to evaluate the postoperative course after administration of the drug. RESULTS: Seventy-three patients received inhaled milrinone from June 2002 to February 2005. Mean age was 64+/-13 years, with a mean preoperative Parsonnet score of 27+/-14. Inhaled milrinone (5mg) was administered before (n=30) or after (n=40) CPB, three patients had off-pump procedures and were excluded. CPB time was 145+/-78 min with cross-clamping times of 91+/-56 min without any significant difference between groups. Fifty-four patients (74%) had difficult separation from CPB, 14 patients (19%) required an intra-aortic balloon pump and 10 patients (14%) needed emergency reinitiation of CPB for hemodynamic instability. Ten patients died in the perioperative period (13.7%). Patients receiving inhaled milrinone prior to CPB initiation had a lowering pulmonary artery pressure after CPB (p<.01) and had less emergency reinitiation of CPB after weaning (3% vs 23%, p=.02) as compared to those with administration after CPB. No detectable side effects were directly linked to the administration of the drug. CONCLUSION: In this high-risk cohort, use of inhaled milrinone was well tolerated. Administration before initiation of CPB could help weaning from CPB. PMID- 17400469 TI - Immunohistochemistry-detected microscopic tumor spread affects outcome in en-bloc resection for T3-chest wall lung cancer. AB - OBJECTIVE: This study is aimed at analyzing the effect of immunohistochemistry detected microscopic tumor spread on long-term survival after en-bloc lung and chest wall resection for T3-chest wall non-small cell lung cancer (NSCLC). METHODS: We retrospectively reviewed 47 patients (mean age 64.4+/-7.1 years, range 48-77) who underwent radical en-bloc lung and chest wall resection for NSCLC between 1987 and 2000. Resection margins, invasion depth, and lymph nodes were re-assessed by immunohistochemistry with AE1/AE3 anti-cytokeratin and anti CEA monoclonal antibodies. RESULTS: Operative mortality and morbidity were 2.1% and 34%, respectively. At immunohistochemistry analysis, five patients (10.6%) revealed microinfiltration of the resection margins that was significantly correlated with the development of local recurrence (p<0.005). Nodal micrometastases were found in 4 out of 33 N0 patients (12.1%), and correlated with distant relapse (p<0.001). Overall and disease-free survivals were significantly influenced by N-status (p<0.001), especially after re-evaluation of micrometastases (p<0.0001), and resection margins microinfiltration (p<0.0001) being these last two the only significant prognostic factors at Cox regression analysis. Five-year overall survival in radically resected patients was 73%. CONCLUSIONS: In this study immunohistochemical analysis allowed to identify patients at higher risk of recurrence following en-bloc resection for T3-chest wall NSCLC. PMID- 17400470 TI - Aortic translocation for transposition of great arteries and left ventricular outflow tract obstruction. PMID- 17400471 TI - Virus-mediated modulation of the host endocrine signaling systems: clinical implications. AB - Viruses, which are among the simplest infective pathogens, can produce characteristic endocrine manifestations in infected patients. In addition to the classic modification of the host endocrine system by either direct or indirect destruction of the endocrine organs and/or effects exerted by systemic production of inflammatory and/or stress mediators, recent progress in molecular virology and endocrinology has revealed that virus-encoded molecules might alter the host endocrine-signaling systems by affecting extracellular and/or intracellular signal transduction and hormone sensitivity of host target tissues. Here, we provide a brief overview of such viral-mediated modulation of host endocrine signaling systems. We propose that virus-encoded molecules and the signaling systems they influence are potential therapeutic targets for the treatment of disorders that are associated with some viral infections. PMID- 17400473 TI - Selection of spiking materials for studies on the clearance of agents of transmissible spongiform encephalopathy during plasma fractionation. AB - Manufacturers of plasma derivatives are encouraged to perform product-specific investigational studies on the ability of their processes to remove prion agents. Such studies are invariably performed using spiking materials derived from infected brain tissue. However, there is little guidance available on which preparations are suitable and no consensus on the acceptability of resultant data. Further research is required to resolve this question. PMID- 17400472 TI - Molecular mechanisms for adaptive tolerance and other T cell anergy models. AB - Since the original description of T cell anergy in CD4 clones from mice and humans, a number of different unresponsive states have been described, both in vivo and in vitro, that have been called anergic. While initial attempts were made to understand the similarities between the different models, it has now become clear from biochemical experiments that many of them have different molecular mechanisms underlying their unresponsiveness. In this review we will detail our own work on the in vivo model referred to as adaptive tolerance and then attempt to compare this biochemical state to the multitude of other states that have been described in the literature. PMID- 17400474 TI - Japanese encephalitis virus production in Vero cells with serum-free medium using a novel oscillating bioreactor. AB - A novel oscillating bioreactor, BelloCell, was successfully applied for the cultivation of Vero cells using serum-free medium, and the production of Japanese encephalitis virus. The BelloCell requires no air sparging, pumping, or agitation, and thus provides a low shear environment. Owing to its simple design, BelloCell is extremely easy to handle and operate. Using this BelloCell (500 ml culture), Vero cells reached a maximum number of 2.8 x 10(9) cells and the Japanese encephalitis virus yield reached 6.91 x 10(11) PFU, versus 9.0 x 10(8) cells and 2.98 x 10(11) PFU using a spinner flask (500 ml) with microcarriers. The cell yield and virus production using BelloCell were markedly higher than with microcarrier culture. The neutralizing capacity of the Japanese encephalitis virus produced using BelloCell was equal to that using a microcarrier system. Therefore, these benefits should enable BelloCell to be adopted as a simple system for high population density cell culture and virus production. PMID- 17400475 TI - Octomeric pyruvate-ferredoxin oxidoreductase from Desulfovibrio vulgaris. AB - Pyruvate-ferredoxin oxidoreductatse (PFOR) carries out the central step in oxidative decarboxylation of pyruvate to acetyl-CoA. We have purified this enzyme from Desulfovibrio vulgaris Hildenborough (DvH) as part of a systematic characterization of as many multiprotein complexes as possible for this organism, and the three-dimensional structure of this enzyme has been determined by a combination of electron microscopy (EM), single particle image analysis, homology modeling and computational molecular docking. Our results show that the 1MDa DvH PFOR complex is a homo-octomer, or more precisely, a tetramer of the dimeric form of the related enzyme found in Desulfovibrio africanus (Da), with which it shares a sequence identity of 69%. Our homology model of the DvH PFOR dimer is based on the Da PFOR X-ray structure. Docking of this model into our 17A resolution EM reconstruction of negatively stained DvH PFOR octomers strongly suggests that the difference in oligomerization state for the two species is due to the insertion of a single valine residue (Val383) within a surface loop of the DvH enzyme. This study demonstrates that the strategy of intermediate resolution EM reconstruction coupled to homology modeling and docking can be powerful enough to infer the functionality of single amino acid residues. PMID- 17400476 TI - Integrating electron microscopy information into existing Distributed Annotation Systems. AB - The increase of daily released bioinformatic data has generated new ways of organising and disseminating information. Specifically, in the field of sequence data, many efforts have been made not only to store information in databases, but also to annotate it and then share these annotations through a standard XML (eXtensible Markup Language) protocol and appropriate integration clients. This is the context in which the Distributed Annotation System (DAS) has emerged in genomics. Additionally, initiatives in the field of structural data, such as the extension of DAS to atomic resolution data, which generated the SPICE client, have also occurred. This paper presents 3D-EM DAS, a further extension of the DAS protocol that allows sharing annotations about hybrid models. This annotation system has been built on the basis of the EMDB, which stores Three-dimensional Electron Microscopy (3D-EM) volumes, PDB, which houses atomic coordinates, and UniProt (for protein sequences) databases. In this way, annotations for sequences, atomic coordinates, and 3D-EM volumes are collected and displayed through a single graphical visualization client. Thus, users have an integrated view of all the annotations together with the whole macromolecule (3D-EM map coming from EMDB), the atomic resolution structures fitted into it (coordinates coming from PDB) and the sequences corresponding to each of the structures (from UniProt). PMID- 17400477 TI - Abnormalities of language networks in temporal lobe epilepsy. AB - Functional magnetic resonance imaging (fMRI) in patients with temporal lobe epilepsy (TLE) has demonstrated reorganisation of language functions with greater involvement of the non-dominant hemisphere. The structural brain connections supporting this atypical language dominance have not previously been identified. We performed fMRI of language functions and imaging of white matter connections using MR tractography in 14 patients with unilateral TLE and hippocampal sclerosis and 10 controls. Verb generation and reading comprehension paradigms were used to define functional regions which were used to generate starting regions for tractography. Controls and right TLE patients had a left-lateralised pattern of both language-related activations and the associated structural connections. Left TLE patients showed more symmetrical language activations, along with reduced left hemisphere and increased right hemisphere structural connections. Subjects with more lateralised functional activation had also more highly lateralised connecting pathways. We provide evidence for structural reorganisation of white matter tracts that reflects the altered functional language lateralisation in left TLE patients. The combination of fMRI and tractography offers a promising tool for studying the reorganisation of language functions in many neurological conditions and may prove useful in predicting language deficits following temporal lobe surgery. PMID- 17400478 TI - The oxygenation response to functional stimulation: is there a physiological meaning to the lag between parameters? AB - To investigate the regulation of the hemodynamic response to functional stimulation, functional near-infrared spectroscopy (fNIRS) has been used, due to its ability to assess the dynamics of oxygenated, deoxygenated and total hemoglobin concentration ([oxy-Hb], [deoxy-Hb] and [tot-Hb]). Concerning the latency of these parameters, recent studies have returned a consistent picture when comparing the oxygenation response in the sensorimotor to the visual system: changes in [oxy-Hb] lead those in [deoxy-Hb] by 1.6+/-0.2 s (mean+/-SD) for the sensorimotor system but not for the visual system (0.1+/-0.3 s). A number of physiological differences between these cortical areas may account for such a discrepancy, however, the methodological properties of transcranial NIRS also have a relevant influence. Here we show that for the motor system the latency between changes in oxy- compared to deoxy-Hb vanishes once efforts are made to reduce the effects of a systemic response accompanying sensorimotor activity. We apply two independent approaches to reduce the systemic response and find a simultaneous change in [oxy-Hb] and [deoxy-Hb] even in response to a motor paradigm. The two approaches are: (i) an experimental paradigm with alternating contralateral and ipsilateral motor performance without interspersed rest periods designed to minimize systemic changes and (ii) a global correction scheme in an experiment, comparing a unilateral motor performance to rest. These data shed some doubt on the alleged fundamental physiological difference between cortical hemodynamic regulation in motor and visual cortex and highlight the relevance to respect contributions of the systemic hemodynamics. PMID- 17400479 TI - Dynamic discrimination analysis: a spatial-temporal SVM. AB - Recently, pattern recognition methods (e.g., support vector machines (SVM)) have been used to analyze fMRI data. In these applications the fMRI scans are treated as spatial patterns and statistical learning methods are used to identify statistical properties of the data that discriminate between brain states (e.g., task 1 vs. task 2) or group of subjects (e.g., patients and controls). We propose an extension of these approaches using temporal embedding. This makes the dynamic aspect of fMRI time series an explicit part of the classification. The proposed pattern recognition approach uses both spatial and temporal information. Temporal embedding was implemented by defining spatiotemporal fMRI observations and applying a support vector machine to these temporally extended observations. This produces a discriminating weight vector encompassing both voxels and time. The resulting vector furnishes discriminating responses, at each voxel without imposing any constraints on their temporal form. PMID- 17400480 TI - Attention and reality constraints on the neural processes of empathy for pain. AB - Recent brain imaging studies have shown that the neural substrates underlying the ability to infer and share the feeling of pain of other individuals overlap with the pain matrix that mediates the process of one's own pain. While there has been evidence that the neural activity mediating pain experience is influenced by top down attention, it remains unclear whether the neural substrates of empathy for pain are modulated by top-down controlled mechanisms. The current work investigated whether the neural correlates of empathic processes of pain are altered by task demand and prior knowledge of stimulus reality. Subjects were scanned using functional magnetic resonance imaging (fMRI) while watching pictures or cartoons of hands that were in painful or neutral situations. Subjects were asked either to evaluate pain intensity supposedly felt by the model or to count the number of hands in the stimulus displays. Relative to counting neutral stimuli, rating pain intensity of painful pictures and cartoons induced increased activation in ACC/paracingulate and the right middle frontal gyrus. Rating pain intensity also activated the inferior frontal cortex bilaterally and the right insula/putamen for pictures but activated the left parietal cortex, the postcentral gyrus, and the occipito-temporal cortex for cartoons. However, the neural activities related to pain rating were eliminated when subjects counted the number of hands in the painful stimuli. In addition, the ACC activity associated with empathy for pain was stronger for the pictures than for the cartoons. Our findings indicate that the involvement of the neural substrates underlying pain-related empathy is constrained by top-down attention and contextual reality of stimuli. PMID- 17400481 TI - Phylogenetic and taxonomic incongruence involving nuclear and mitochondrial markers in Korean populations of the freshwater snail genus Semisulcospira (Cerithioidea: Pleuroceridae). AB - We investigated the population genetic structure and phylogenetic relationships of four morphospecies of Semisulcospira sampled from multiple South Korean drainages. One, S. extensa, displayed modest levels of genetic diversity and formed a species-specific clade for both mitochondrial (mt) and nuclear markers. In contrast, the other three, S. coreana, S. gottschei, and S. libertina, were polyphyletic for both mt and nuclear markers. They formed, together with other nominal congeners (S. forticosta, S. multicincta, S. nodiperda, and S. tegulata), a taxonomically heterogeneous species complex containing population-level admixtures of genotypes from numerically predominant modal clades in addition to rare, phylogenetically divergent, mt and nuclear genotypes. The modal mt haplotypes exhibited far more geographic than taxonomic congruence and typically co-clustered into drainage-specific tip clades, irrespective of nominal taxonomic status. The evolutionary origins of the observed non-extensa phylogenetic heterogeneity are unclear at present although the available data do not support paralogous mt marker phenomena or the presence of cryptic species. We cannot distinguish among retention of ancestral polymorphisms or recticulate evolutionary origins as explanatory mechanisms and it may require the genetic characterization of Semisulcospira lineages throughout much of their collective east Asian range to address this issue. Based on the available data, we consider it best to view all of our non-extensa Korean study taxa as a single polymorphic species complex of the type species S. libertina. There is considerable evidence that similarly complex genetic structuring, at least for mt, may be typical of many other freshwater cerithioidean snail taxa. In light of our Semisulcospira results, we suggest that meaningful molecular phylogenetic characterization of freshwater cerithioidean lineages may require the use of both mt and nuclear markers together with population level sampling of all nominal taxa within regional drainages. PMID- 17400482 TI - Molecular phylogeny of the scincid lizards of New Caledonia and adjacent areas: evidence for a single origin of the endemic skinks of Tasmantis. AB - We use approximately 1900bp of mitochondrial (ND2) and nuclear (c-mos and Rag-1) DNA sequence data to recover phylogenetic relationships among 58 species and 26 genera of Eugongylus group scincid lizards from New Caledonia, Lord Howe Island, New Zealand, Australia and New Guinea. Taxon sampling for New Caledonian forms was nearly complete. We find that the endemic skink genera occurring on New Caledonia, New Zealand and Lord Howe Island, which make up the Gondwanan continental block Tasmantis, form a monophyletic group. Within this group New Zealand and New Zealand+Lord Howe Island form monophyletic clades. These clades are nested within the radiation of skinks in New Caledonia. All of the New Caledonian genera are monophyletic, except Lioscincus. The Australian and New Guinean species form a largely unresolved polytomy with the Tasmantis clade. New Caledonian representatives of the more widespread genera Emoia and Cryptoblepharus are more closely related to the non-Tasmantis taxa than to the endemic New Caledonian genera. Using ND2 sequences and the calibration estimated for the agamid Laudakia, we estimate that the diversification of the Tasmantis lineage began at least 12.7 million years ago. However, using combined ND2 and c mos data and the calibration estimated for pygopod lizards suggests the lineage is 35.4-40.74 million years old. Our results support the hypothesis that skinks colonized Tasmantis by over-water dispersal initially to New Caledonia, then to Lord Howe Island, and finally to New Zealand. PMID- 17400483 TI - Modulation of articular chondrocyte proliferation and anionic glycoconjugate synthesis by glucosamine (GlcN), N-acetyl GlcN (GlcNAc) GlcN sulfate salt (GlcN.S) and covalent glucosamine sulfates (GlcN-SO4). AB - OBJECTIVES: To investigate, in chondrocyte cultures under conditions for maximizing responses in proliferation and proteoglycan (PG) synthesis, the effects of glucosamine hydrochloride (GlcN.HCl) and glucosamine sulfate (GlcN.S) salts, N-acetyl glucosamine (GlcNAc), and covalently substituted GlcN X,Y,Z(SO(4))(n) (general formula). METHODS: Bovine articular chondrocytes (BAC) were studied under anchorage-independent (AI, alginate beads) and anchorage dependent (AD, plastic surface) conditions. Differentiation markers were evaluated (e.g., cartilage-specific (V+C)(-) fibronectin). Varying concentrations of GlcN.HCl, GlcN.S, GlcNAc and GlcN sulfated at positions -2, -3, -6, (-2,3), ( 3,6) and (-3,4,6), were tested. Cell proliferation, DNA synthesis and [(35)S] sulfate incorporation into newly synthesized PG were determined. RESULTS: Increasing GlcN.HCl or GlcN.S concentrations gave decreasing net PG synthesis. Compounds showed more pronounced effects in AD cultures (expressing the V(-)C(+) fibronectin isoform) compared to AI cultures ((V+C)(-) isoform). Addition of GlcN.HCl or GlcN.S gave a concentration-dependent decrease in BAC proliferation, partially prevented by glucose (Glc). GlcNAc was not inhibitory. Addition of GlcN 2-SO(4) or GlcN-2,6-diSO(4) did not affect proliferation or DNA synthesis. The other GlcN-sulfates gave varying inhibitory effects, which for GlcN-3-SO(4) were reversed by inosine. CONCLUSIONS: The free amino group of GlcN seems responsible for inhibition of chondrocyte proliferation and PG synthesis. These effects were greater under higher concentrations of GlcN in AD vs AI conditions. GlcN.HCl behaves similarly to GlcN.S, but differential effects with GlcN-X,Y,Z(SO(4))(n) isomers were observed. Acetylation or sulfation of the GlcN amino group reverses or partially reverses, respectively, anti-proliferative effects of GlcN. Sulfation of GlcN, at positions 3 and 6 results in complex effects on AC proliferation and PG synthesis. PMID- 17400484 TI - The role of the agranular insular cortex in anticipation of reward contrast. AB - Sixteen male Long-Evans rats were tested on a modified version of Flaherty et al.'s [Flaherty, C. F., Turovsky, J., & Krauss, K. L. (1994). Relative hedonic value modulates anticipatory contrast. Physiology and Behavior, 55, 1047-1054.] anticipatory contrast paradigm to assess memory for the anticipation of reward. Prior to testing each rat received either a control or quinolinic acid induced lesion of the agranular insular cortex. In the home cage, each rat was allowed to drink a water solution containing 2% sucrose for 3 min followed by a water solution containing 32% sucrose for 3 min. Across 10 days of testing, the control rats showed significantly increased anticipatory discriminability as a function of days. In contrast, rats with agranular insular cortex lesions failed to show anticipatory discriminability. The results of a preference task revealed that both groups could perceptually discriminate between a 2% and a 32% sucrose solution. The data suggest that the agranular insular cortex may be involved in the anticipation of reward. PMID- 17400485 TI - Cardiac risk reduction in patients with intermittent claudication. PMID- 17400486 TI - Dacron or PTFE for above-knee femoropopliteal bypass. a multicenter randomised study. AB - OBJECTIVES: To compare polytetrafluorethylene (PTFE) and polyester grafts (Dacron) for above knee femoropopliteal bypass. DESIGN: Multicenter randomised clinical trial. MATERIAL AND METHODS: 427 patients were randomised between 6mm Dacron (Uni-Graft, B. Braun Melsungen AG, 34212 Melsungen, Germany) and PTFE (Goretex, W. L. Gore & Ass. Inc., Newark DE 19711, USA) above-knee femoropopliteal bypass grafts within 13 centres in Denmark (n=261), Norway (n=113) & Finland (n=53) between 1993 and 1998. Fourteen (3%) patients were excluded, leaving 413 patients with 208 Dacron and 205 PTFE grafts for analysis. Age, gender, indication (claudication: 65%), run-off (2 or 3 vessels: 76%), diabetes (17%) and hypertension (31%) as well as cerebrovascular (9%) and cardiac (33%) risks were evenly distributed. Patients were followed postoperatively at 1, 12 and 24 months. Patency assessment was based on ankle-brachial pressures or imaging in case of doubt. RESULTS: The two-year primary patency rates for Dacron and PTFE were 70% and 57% (p=0.02), whereas the secondary patency rates were 76% and 65% (p=0.04), respectively. Primary patency at two years was significantly influenced by the number of patent crural vessels (two or three 67%, one 50%, p=0.01). Amputations at two years, major in 4% and minor in 3%, 30-days mortality and complications (wound infections: 3% and other wound complications: 13%) occurred equally frequent in both groups. At two years, patients treated for critical limb ischemia had a major amputation more often than patients operated on for intermittent claudication, 10 and 3 respectively (p=0.003), and had higher mortality rates, 20% and 8% respectively (p=0.001). CONCLUSION: This trial confirms that Dacron is at least as durable as PTFE for above-knee bypass procedures, and might even be superior. PMID- 17400487 TI - A multicenter review of carotid body tumour management. AB - OBJECTIVE: Carotid body tumour (CBT) is a rare but the most common form of head and neck paraganglioma (PGL). We present the biggest ever series on CBT in UK/EU discussing diagnostic challenges, surgical treatment and complications of surgical intervention. METHOD: A detailed proforma was designed and sent to all members of Joint Vascular Research Group (JVRG). Data of 95 patients was collected. Generic terms including carotid body tumour/s, or paraganglioma/s were used to search a variety of electronic database in order to get latest informations available in literature. RESULTS: A total of 95 patients were recorded in our data from 1979 to 2005. Mean age of presentation was 55 years. Incidence was higher in females. CBT was more common on right side (58%). 18% tumours were bilateral. Neck lump (98%) and pressure symptoms including cranial nerve deficits and pain were main presenting complaints. About 18% of tumours were familial. Only 4.2% were malignant. Duplex scan is the best investigation for diagnosis, though MRI, DSA and CT scan are important for preoperative assessment. Surgery is the treatment of choice. Stroke and cranial nerve injury constitute postoperative morbidity (35%) and mortality (1%). Incidence of postoperative cranial nerve deficit was about 19%. Combined ipsilateral and contralateral recurrence rate was 4.2%. CONCLUSION: CBT is a rare condition which needs surgical excision by experienced vascular surgeon. Surgical resection is associated with significant morbidity of 35% and mortality of 1%. Mostly CBT is benign but malignant forms are not uncommon. PMID- 17400488 TI - Detection of 28 novel mutations in the Wiskott-Aldrich syndrome and X-linked thrombocytopenia based on multiplex PCR. AB - The Wiskott-Aldrich syndrome (WAS) is an X-linked disorder including microthrombocytopenia, eczema and immunodeficiency. A mild form is known as the X linked thrombocytopenia (XLT). We screened 150 individuals or families based on a multiplex PCR method. We found 28 novel mutations (7 missense, 1 nonsense, 1 nonstop change, 5 splice site mutations and 14 deletions or insertions). The method relied on the co-synthesis of 5 amplicons and direct sequencing, optimizing the novel protocol proposed by Jones et al. [L.N. Jones, M.I. Lutskiy, J. Cooley, et al. A novel protocol to identify mutations in patients with Wiskott Aldrich syndrome, Blood Cells Mol. Dis. 28 (2002) 392-398]. It was thus possible to identify faster and at a lower cost the mutations in newly diagnosed patients. The mutation distribution, according to the type, was in keeping with the distribution reported previously. No clear-cut genotype-phenotype correlation was observed. PMID- 17400489 TI - The immune status of the bovine uterus during the peripartum period. AB - The post-partum period in cattle is characterised by an increased risk of infection of the uterus, as the anatomical barriers are broached during parturition and remain open for several days. Infection of the uterus is largely influenced by the balance between bacterial contamination and the local and systemic immune status during pregnancy and around parturition. Infectious diseases are more prevalent during this period, because of an impaired immune status before and immediately after parturition. Neutrophils play a primary role in the defence of the uterus against infection. Influx of neutrophils into the uterus is thought to be mediated by chemoattractants, chemokines and adhesion molecules, such as beta2-integrin (complement receptor 3) and L-selectin (CD62L). Other cellular components activated in the uterus during this period include lymphocytes, eosinophils, mast cells and macrophages. The major classes of immunoglobulins (IgM, IgA and IgG), either by passive diffusion or local production, play an important protective role in the uterus by acting as opsonins to enhance phagocytosis, stimulating the complement pathways or blocking pathogens from adhering to mucosal surfaces. Endometrial cells express toll-like receptor 4 (TLR4), which recognises lipopolysaccharides of Escherichia coli and other Gram negative bacteria, the most common causes of bovine endometritis. Activation of TLR4 triggers the production of tumour necrosis factor alpha and other pro-inflammatory cytokines. The periparturient period is also characterised by an increased secretion of prostaglandin F(2alpha), which enhances uterine immune defences. PMID- 17400490 TI - Diagnostic criteria for CRPS I: differences between patient profiles using three different diagnostic sets. AB - Complex Regional Pain Syndrome type I (CRPS I) is an illness which usually occurs due to major or minor tissue injury to the extremities. Because a unique pathophysiological mechanism for CRPS I has not yet been established, the diagnosis is based on observation and measurement of clinical symptoms and signs. In this study, a comparison was made between three sets of diagnostic criteria (the IASP, Bruehl et al. and Veldman et al.) based on patient reports and physicians' assessments of signs and symptoms associated with CRPS I, in 372 outpatients suspected of having CRPS I. Agreement between CRPS I diagnosis among the three sets was poor (kappa-range: 0.29-0.42), leading to positive CRPS I diagnoses according to Veldman et al.'s criteria in 218 cases (59%), according to the IASP in 268 cases (72%), and according to Bruehl et al. in 129 cases (35%). Significant differences in patient profiles were found between the diagnostic sets for the number of patients reporting continuing disproportionate pain, larger area affected than the initial trauma (both p<0.001), increase of symptoms due to exercise (p=0.009), edema (p=0.015), temperature asymmetry (p=0.015), hyperesthesia, allodynia (both p<0.001) and hyperalgesia (p=0.036). Similarly, significant differences emerged for physicians' observations of hyperesthesia and allodynia (both p<0.001). Highest combined values of sensitivity (SE) and specificity (SP) for the strongest cases of presence (n=108) or absence (n=62) of CRPS I were found for reported hyperesthesia (SE+SP:165%), allodynia (160%), observed color asymmetry (162%), hyperesthesia (157%), temperature asymmetry (154%) and edema (152%). The lack of agreement between the different diagnostic sets for CRPS I and the different clinical profiles that result from it may lead to different therapeutic and study populations, hampering adequate treatment and scientific development for this illness. We propose explicit reference to diagnostic criteria used in studies, and registration in trials of a broad variety of CRPS I features, as used in this study, to make subgroup phenotyping and post hoc analyses based on different diagnostic criteria possible. PMID- 17400491 TI - Predictive factors of chronic pain and disability in whiplash: a Delphi poll. AB - The purpose of this study was to establish consensus on what factors might predict chronic pain and disability in whiplash injuries following motor vehicle collisions. A Delphi poll involving two rounds of data collection was used as a way to reach consensus among participating experts. Participating experts identified several physical, historical, psychological, and behavioral response factors that they believed were related to the development of chronic pain and related disability in people who have developed whiplash-associated disorders. These included: (1) prior history: previous history of chronic pain; (2) physical factors: high severity of injury, presence of constant neck pain since the accident; (3) psychological factors: belief that pain mean danger and that he or she should avoid exercise, tendency to somatize, to have positive thoughts about pain, catastrophic thinking, low self-efficacy expectancies, and to have symptoms of posttraumatic stress disorders; and (4) behavioral responses to symptoms: avoidance of activities that involve moving the neck through fear of experiencing more pain, and restriction in the use and movement of the neck. These findings may be used to help identify the specific domains that should be assessed in studies seeking to predict which individuals are at risk to develop chronic pain and disability following initial whiplash-associated disorders sustained in crash. If these results are supported by future studies, then they could be used to help develop intervention programs that could prevent long-term pain and disability in whiplash patients who are considered to be at risk to develop chronic disabling pain problems. PMID- 17400492 TI - Diagnostic criteria and follow-up parameters in complex regional pain syndrome type I--a Delphi survey. AB - BACKGROUND: Although the current clinical guideline of diagnostic criteria for the complex regional pain syndrome I (CRPS I) is a landmark endeavour to define this complex condition it does not prioritise its most important clinical manifestations. AIM: We set out to obtain an expert agreed priority list of diagnostic and follow-up parameters in the diagnosis and management of CRPS I. METHODS: A two round Delphi survey: We asked international experts to list (first round) and weight (second round) parameters (scale 1-10) they believed to be relevant in diagnosis and follow-up. Median ratings and interquartile ranges (IQR) were calculated. Rates 7 and IQR 3 depicted important and expert agreed parameters. RESULTS: Thirty-two diagnostic and 23 follow-up listings and ratings of 13 experts were available for analysis. In three domains (clinical presentation, further examinations and follow-up) experts agreed on the following parameters, pain (10; 9-10) with its subcategories hyperesthesia (7; 5-8) hyperalgesia (8; 8-8) and allodynia (8; 7-10), signs with oedema (9; 8-10) and colour change (8; 5-8) and mobility with its categories motor change (7; 5-8) and decreased range of motion (8; 8-8). The experts agreed that no further examinations were necessary for diagnosis (10; 8-10). The agreed important follow up parameter was clinical course (10; 8-10) with its categories decrease in pain (8; 8-9) and hyperalgesia (8; 6-8), decreased oedema (8; 7-10) and improvements in motor function (10; 8-10) and strength (8; 6-9). CONCLUSION: This expert survey conveys an agreed set of relevant diagnostic parameters of CRPS I and proposes that in follow-up examinations treatment success should be based on restoration of those manifestations. PMID- 17400493 TI - Measuring nanopore size from the spin-lattice relaxation of CF4 gas. AB - The NMR 19F spin-lattice relaxation time constant T1 for CF4 gas is dominated by spin-rotation interaction, which is mediated by the molecular collision frequency. When confined to pores of approximately the same size or smaller than the bulk gas mean free path, additional collisions of molecules with the pore walls should substantially change T1. To develop a method for measuring the surface/volume ratio S/V by measuring how T1 changes with confinement, we prepared samples of known S/V from fumed silica of known mass-specific surface area and compressed to varying degrees into cylinders of known volume. We then measured T1 for CF4 in these samples at varying pressures, and developed mathematical models for the change in T1 to fit the data. Even though CF4 has a critical temperature below room temperature, we found that its density in pores was greater than that of the bulk gas and that it was necessary to take this absorption into account. We modeled adsorption in two ways, by assuming that the gas condenses on the pore walls, and by assuming that gas in a region near the wall is denser than the bulk gas because of a simplified attractive potential. Both models suggested the same two-parameter formula, to which we added a third parameter to successfully fit the data and thus achieved a rapid, precise way to measure S/V from the increase in T1 due to confinement in pores. PMID- 17400494 TI - Evaluation of the influence of housefly maggot meal (magmeal) diets on catalase, glutathione S-transferase and glycogen concentration in the liver of Oreochromis niloticus fingerling. AB - Influence of housefly maggot meal (magmeal) diets on the activities of catalase (CAT), glutathione S-transferase (GST) and glycogen concentration in liver of Tilapia Oreochromis niloticus fingerling was evaluated. Triplicate groups of fifteen fish (initial average weight 2.0+/-0.1 g) were fed eight weeks with seven test diets (in average 36% crude protein, dry matter) formulated by replacing fish meal with magmeal. Percentage body weight gain (591-724.46%), food conversion ratio (1.05-1.22) and standard growth rate (3.45-3.76) in all feeding groups were not significantly different (P<0.05). No significant difference (P<0.05) was observed in liver glycogen reserve (175.27-236.88 micromol g(-1)) among the fish groups. Hepatic catalase activity also did not differ significantly. However, elevated glutathione S-transferases activities were observed when fish received higher dietary magmeal concentration. This might have been temporary with no real physiological implication when appraised by the growth responses. These results indicate that magmeal was well utilized by the fish and its incorporation into tilapia diets seems to have no oxidative stress generating effect on fish metabolism and may not be containing any compound that stimulates the generation of reactive oxygen species. Magmeal can effectively be used as an alternative protein source in tilapia fingerling production. PMID- 17400495 TI - The expression of AmphiTCTP, a TCTP orthologous gene in amphioxus related to the development of notochord and somites. AB - The translationally controlled tumor protein (TCTP) is highly conserved and has been widely found in eukaryotic organisms. Here, we report the phylogenetic analysis and developmental expression of AmphiTCTP, a TCTP homologous gene in cephalochordate amphioxus. Phylogenetic analysis indicates that the putative protein of AmphiTCTP is close to its vertebrate orthologs. The mRNA of AmphiTCTP is found in fertilized eggs, early cleavage embryo and most of the early developmental stages by in situ hybridization and RT-PCR, but its expression is not detectable from late cleavage stage to mid-gastrula. The expression of AmphiTCTP in zygotes and early cleavage stages shows that AmphiTCTP may be a maternal gene. From the early neurula stage onward, AmphiTCTP transcript is localized in the presumptive notochord, presomitic mesoderm, and nascent somites. However, its expression is gradually down-regulated after the notochord and somites have been formed. The expression pattern of AmphiTCTP thus coincides with the differentiation of the notochord and somites, this suggests that AmphiTCTP may not be a housekeeping gene and may play an important role in mesoderm development. PMID- 17400496 TI - Identification of a novel DNA methyltransferase 2 from the brine shrimp, Artemia franciscana. AB - DNA methyltransferase 2 (Dnmt2) is a dual-specificity DNA methyltransferase, which contains a weak DNA methyltransferase and novel tRNA methyltransferase activity. However, its biological function is still enigmatic. To elucidate the expression profiles of Dnmt2 in Artemia franciscana, we isolated the gene encoding a Dnmt2 from A. franciscana and named it as AfDnmt2. The cDNA of AfDnmt2 contained a 1140-bp open reading frame that encoded a putative Dnmt2 protein of 379 amino acids exhibiting 32% approximately 39% identities with other known Dnmt2 homologs. This is the first report of a DNA methyltransferase gene in Crustacean. By using semi-quantitative RT-PCR, AfDnmt2 was found to be expressed through all developmental stages and its expression increased during resumption of diapause cysts development. Southern blot analysis indicated the presence of multiple copies of AfDnmt2 genes in A. franciscana. PMID- 17400497 TI - Proteolytic activity and protease classes in the zooplankton species Calanus finmarchicus. AB - The temperature optimum for the general proteolytic activity in the crude extract of Calanus finmarchicus was 50 degrees C and the pH optimum was found to be 7. The use of specific protease inhibitors resulted in the identification of at least three protease classes in the crude extract of C. finmarchicus. Those classes were serine, metallo and aspartic proteases. The serine and metallo proteases were found to be dominant under neutral to alkaline conditions and the aspartic proteases were dominant under acidic conditions. The cysteine proteases, if at all present, did not seem to be active or present in any substantial amount. The data presented points towards controlling and preserving the protein content in fresh C. finmarchicus. PMID- 17400498 TI - Chronic intracerebroventricular TLQP-21 delivery does not modulate the GH/IGF-1 axis and muscle strength in mice. AB - OBJECTIVE: Biallelic ablation of VGF determines a dwarf phenotype. VGF precursor protein encodes for different biologically active peptides none of which has been related to growth or muscular abnormalities. Here we present the first attempt to fill this gap. We tested the hypothesis that a recently identified VGF-derived peptide, TLQP-21, shown to centrally modulate metabolic functions, could also modulate growth hormone (GH)-axis and muscle strength. DESIGN: Adult male mice were chronically icv injected with TLQP-21 (15 microg/day for 14 days). Physiological, molecular and behavioral parameters related to the GH/IGF-1-axis were investigated. RESULTS: Except for a reduction in the soleus weight, TLQP-21 did not affect GH/IGF-1-axis mediators, muscle strength and muscle weight. CONCLUSIONS: Results collected exclude a role for TLQP-21 in modulating the GH/IGF1-axis and muscle functions. VGF-derived peptides involved in the dwarf phenotype of VGF-/- mice have to be identified yet. PMID- 17400499 TI - Metabolic flux analysis in a nonstationary system: fed-batch fermentation of a high yielding strain of E. coli producing 1,3-propanediol. AB - Metabolic fluxes estimated from stable-isotope studies provide a key to understanding cell physiology and regulation of metabolism. A limitation of the classical method for metabolic flux analysis (MFA) is the requirement for isotopic steady state. To extend the scope of flux determination from stationary to nonstationary systems, we present a novel modeling strategy that combines key ideas from isotopomer spectral analysis (ISA) and stationary MFA. Isotopic transients of the precursor pool and the sampled products are described by two parameters, D and G parameters, respectively, which are incorporated into the flux model. The G value is the fraction of labeled product in the sample, and the D value is the fractional contribution of the feed for the production of labeled products. We illustrate the novel modeling strategy with a nonstationary system that closely resembles industrial production conditions, i.e. fed-batch fermentation of Escherichia coli that produces 1,3-propanediol (PDO). Metabolic fluxes and the D and G parameters were estimated by fitting labeling distributions of biomass amino acids measured by GC/MS to a model of E. coli metabolism. We obtained highly consistent fits from the data with 82 redundant measurements. Metabolic fluxes were estimated for 20 time points during course of the fermentation. As such we established, for the first time, detailed time profiles of in vivo fluxes. We found that intracellular fluxes changed significantly during the fed-batch. The intracellular flux associated with PDO pathway increased by 10%. Concurrently, we observed a decrease in the split ratio between glycolysis and pentose phosphate pathway from 70/30 to 50/50 as a function of time. The TCA cycle flux, on the other hand, remained constant throughout the fermentation. Furthermore, our flux results provided additional insight in support of the assumed genotype of the organism. PMID- 17400500 TI - Ethical issues in geriatric feline medicine. AB - Most veterinarians hold a 'pediatric' rather than 'garage mechanic' view of their function. In recent years, sophisticated medical modalities have allowed veterinarians to keep animals alive, and increased value of companion animals in society has increased demand for such treatment. But whereas humans can choose to trade current suffering for extended life, animals seem to lack the cognitive apparatus required to do so. Thus, veterinarians must guard against keeping a suffering animal alive for too long. Clients may be emotionally tied to the animal and blind to its suffering. Part of the veterinarian's role, therefore, is to lead the client to 'recollect' quality of life issues. A second major role for the veterinarian in treating geriatric or chronically ill animals is control of pain and distress. Unfortunately, pain and distress have historically been neglected in both human and veterinary medicine for ideological reasons. It is ethically necessary to transcend this ideology which leads to both bad medicine and bad ethics. PMID- 17400501 TI - [Long-term results and prognostic factors of squamous cell carcinoma of the anal canal treated by irradiation]. AB - PURPOSE: To analyze the prognostic factors of loco regional control (LRC), specific survival (SS) and sphincter conservation (SC) of patients treated by curative and conservative irradiation for an epidermoid cancer of anal canal in our institution. PATIENTS AND METHODS: From 1976 to 2005, 286 patients (pts) were treated by exclusive radiotherapy (180 pts) or chemo-radiotherapy (106 pts) followed by a brachytherapy boost (233 pts) or external beam radiotherapy boost (24 pts). Forty-three pts were stage I, 154 stage II, 31 stage IIIA and 53 stage IIIB. RESULTS: The mean follow-up was 65 months (range: 1.3-250 months). The 5 years-overall survival and SS rates were 66.4% and 78.1% respectively. In multivariate analysis, tumor size (>or=40 mm) [RR=2.1], node involvement (RR=2.4), and poor response (<75%) to first course irradiation [RR=1.9], local relapse (RR=4.5) and distant metastases were factors of poor prognosis for SS. Five-years-LRC were 71.5% (88% for stage I, 69% for stage II, 77%, for stage IIIA and 60% for stage IIIB). Prognosis factors of LCR were tumor size (RR=2.5), response to first course of irradiation (RR=2.9). SC was 71% at 5 years. Prognosis factors of SC were tumor size (RR=1.9) and response to first course of irradiation (RR=2.4). CONCLUSION: The results of this series are similar to those of the literature. As well as initial tumor extension, response to first course of irradiation was found as prognostic factor on LCR, SS, SC. Our results are similar to other series and brachytherapy seems not to be deleterious. Its impact to local control remains to be evaluated. PMID- 17400502 TI - Human gastric glycosphingolipids recognized by Helicobacter pylori vacuolating cytotoxin VacA. AB - Many bacterial toxins utilize cell surface glycoconjugate receptors for attachment to target cells. In the present study the potential carbohydrate binding of Helicobacter pylori vacuolating cytotoxin VacA was investigated by binding to human gastric glycosphingolipids on thin-layer chromatograms. Thereby a distinct binding of the toxin to two compounds in the non-acid glycosphingolipid fraction was detected. The VacA-binding glycosphingolipids were isolated and characterized by mass spectrometry and proton NMR as galactosylceramide (Galbeta1Cer) and galabiosylceramide (Galalpha4Galbeta1Cer). Comparison of the binding preferences of the protein to reference glycosphingolipids from other sources showed an additional recognition of glucosylceramide (Glcbeta1Cer), lactosylceramide (Galbeta4Glcbeta1Cer) and globotriaosylceramide (Galalpha4Galbeta4Glcbeta1Cer). No binding to the glycosphingolipids recognized by the VacA holotoxin was obtained with a mutant toxin with deletion of the 37 kDa fragment of VacA (P58 molecule). Collectively our data show that the VacA cytotoxin is a glycosphingolipid binding protein, where the 37 kDa moiety is required for carbohydrate recognition. The ability to bind to short chain glycosphingolipids will position the toxin close to the cell membrane, which may facilitate toxin internalization. PMID- 17400503 TI - Galleria mellonella as a model host to study infection by the Francisella tularensis live vaccine strain. AB - We used the killing of Galleria mellonella (Lepidoptera: Pyralidae; the greater wax moth) caterpillar by the live vaccine strain (LVS) of Francisella tularensis to develop an invertebrate host system that can be used to study F. tularensis infection and the in vivo effects of antibacterial compounds on F. tularensis LVS. After injection into the insect hemocoel, F. tularensis LVS, killed caterpillars despite the association of LVS with hemocytes. The rate of killing depended on the number of bacteria injected. Antibiotic therapy with ciprofloxacin, levofloxacin or streptomycin administered before or after inoculation prolonged survival and decreased the tissue burden of F. tularensis in the hemocoel. Delayed drug treatment reduced the efficacy of antibacterials and especially streptomycin. The G. mellonella-F. tularensis LVS system may facilitate the in vivo study of F. tularensis, efficacy with antibacterial agents. PMID- 17400504 TI - Paracoccin, an N-acetyl-glucosamine-binding lectin of Paracoccidioides brasiliensis, is involved in fungal growth. AB - Paracoccin is an N-acetyl-glucosamine-binding lectin from Paracoccidioides brasiliensis, which can be obtained in small amounts either from culture supernatants or yeast cell extracts. In the present work, immunoelectron microscopy with mouse anti-paracoccin IgG localized the antigen to the cell wall of P. brasiliensis yeast forms. Paracoccin interacted with chitin, and colocalized with beta-1,4-homopolymer of GlcNAc to the budding sites of P. brasiliensis yeast cell. In order to evaluate the role of paracoccin on fungal growth, yeast cells were cultivated in the presence of anti-paracoccin antibodies. A significant reduction of both colony forming units and individual yeast cells was observed as well as morphological alterations such as smaller colonies and cells more loosely aggregated than in control cultures without the antibody. A role of paracoccin on the cell wall organization was reinforced by alterations in the labeling pattern of chitin when yeasts were treated with anti paracoccin antibodies. Binding of specific antibodies to paracoccin may disrupt the paracoccin/chitin interactions, resulting in the inhibition of P. brasiliensis growth. PMID- 17400505 TI - [Mass neonatal screening using biological testing]. AB - Implementation of a generalized screening program for neonatal diseases obeys precise guidelines. The disease must be severe, recognizable at an early stage, accessible to an effective treatment, detected with a non expansive and widely applicable test and it must represent an important health problem. In case of positive results, treatment or prevention shall be offered immediately and any screening program has to be regularly evaluated. There is in France since 1978 a national screening program that depends on a private association ("Association francaise pour le depistage et la prevention des handicaps de l'enfant") and is supervised by the "Caisse nationale d'assurance maladie" and the "Direction Generale de la Sante". Presently, five diseases are included in the screening program: phenylketonuria, hypothyroidism, congenital adrenal hyperplasia, cystic fibrosis and sickle cell disease, the latter only in at risk newborns. Toxoplasmosis represents a particular problem because screening takes place only in children of mothers that have not been controlled during their pregnancy or in case of seroconversion. Neonatal screening of phenylketonuria and hypothyrodism is unanimously recommended. That of congenital adrenal hyperplasia is approved in most countries. The cases of sickle cell disease and cystic fibrosis are more complex because: 1) all the children that carry the mutations are not affected with a severe disease; 2) there is no curative treatment; 3) parents given information are made anxious, sometimes wrongly if the disease is mild or asymptomatic. The supporters of the screening insist on the interest of an early diagnosis which makes longer the life time of these children, the possibility for the parents to utilize prenatal screening in case of a future pregnancy, and the information given to the heterozygous carriers following a familial screening. The question is raised of the extension of neonatal screening to other diseases. This is now possible due to technical progresses such as the tandem mass spectrometry that can detect about 50 diseases in an only testing. In addition of its cost and of the difficulty to ensure an efficient organization, increasing the number of the screened diseases will raise ethical problems including how the parents will be informed of an incurable disease or a late-onset disease or an entirely asymptomatic disease. It is unanimously admitted that only mendelian diseases should be detected excluding genetic polymorphisms. Analysis of the present situation suggests the following developments: 1) to actualize the guidelines for deciding of a new neonatal screening; 2) to experiment on a local scale any new screening before its extension to the whole country; 3) to create an evaluation committee including paediatricians and epidemiologists and to evaluate on the long term the future of the children; 4) to precisely define the conditions in which the heterozygous carriers will be informed following a familial investigation; 5) to store in a resource biological centre the blood samples in order to utilize this bank for epidemiology studies. PMID- 17400506 TI - Lack of evidence for association of Parkin promoter polymorphism (PRKN-258) with increased risk of Parkinson's disease. AB - Parkin (PRKN) mutations are a common cause of early-onset parkinsonism, however the role of this gene in typical late-onset Parkinson's disease (PD) remains unresolved. A single nucleotide polymorphism in the promoter region (PRKN-258; rs9347683) has been observed to associate with PD, affect age-at-onset (AAO) of symptoms, and to functionally effect differential expression of the PRKN transcript. In the present study, PRKN-258 did not associate with PD, and no evidence for an AAO effect was observed in three age and gender-matched Caucasian patient-control series from Norway, Ireland and the US. These data do not support a role for this common variant in PD etiology. PMID- 17400507 TI - Identification of potential protein interactors of Lrrk2. AB - Pathogenic substitutions in the Lrrk2 protein have been shown to be an important cause of both familial and sporadic parkinsonism. The molecular pathway involved in Lrrk2 dopaminergic neuron degeneration remains elusive. Employing a combination of Lrrk2-mediated protein precipitation and tandem mass spectrometry, we identified 14 potential Lrrk2 binding partners. The majority of these interactions may be subgrouped into three functional cellular pathways: (i) chaperone-mediated response, (ii) proteins associated with the cytoskeleton and trafficking and (iii) phosphorylation and kinase activity. Future investigation of these candidates is now warranted and may help resolve the pathomechanism behind Lrrk2 neurodegeneration. PMID- 17400508 TI - Erythrocyte antioxidant enzyme activities and lipid peroxidation in the erythrocyte membrane of stainless-steel welders exposed to welding fumes and gases. AB - The erythrocyte antioxidant system (superoxide dismutase, SOD; catalase, CAT) and lipid peroxidation (malondialdehyde, MDA) in the erythrocyte membrane were studied in workers continously exposed to welding fumes and gases, which are thought to be oxidant pollutants. Thirty-five welders using the manual metal arc method on stainless steel and 30 controls were studied. Plasma chromium (Cr), manganese (Mn), and cupper (Cu) levels were determined by atomic absorption spectrophotometer (AAS). The erythrocyte antioxidant system activity and lipid peroxidation in the erythrocyte membrane were evaluated. Not only the possible effects of welding fumes but also the effects of smoking were considered. The plasma concentrations of Cr, Mn, and Cu for the exposed welders were significantly higher compared to the control subjects (p<0.001, p<0.01, p<0.001, respectively,). The erythrocyte CAT (p<0.05) and SOD (p<0.05) enzyme activities were significantly higher in the welders but there were not any significant changes in the MDA levels which reflect the lipid peroxidation in the erythrocyte membrane (p>0.05). Smoking has increased the SOD activity in both controls (p<0.05) and welders (p<0.01) and increased the CAT activity in control subjects (p<0.05). Moreover, regardless of smoking, there were some significant correlations between the duration of the exposure to welding fumes and antioxidant defence system (SOD: p<0.05; CAT: p<0.05). The synergistic effects of smoking and other risk factors (welding fumes and gases), which had been shown previously by some clinical data should also be taken into account. As a consequence, the welders should be warned and informed of the synergistic effects of smoking on the adverse effect of welding fumes and gases. PMID- 17400509 TI - Drowning in Australia is more than coastal drowning. PMID- 17400510 TI - Mesenchymal stem cells: a new strategy for immunosuppression? AB - In vitro-generated mesenchymal stem cells (MSCs) initially attracted interest for their ability to undergo differentiation toward cells of different lineages. More recently, a major breakthrough was the discovery that MSCs exert a profound inhibitory effect on T cell proliferation in vitro and in vivo. Subsequently, MSCs were shown also to exert similar effects on B cells, dendritic cells and natural killer cells. These results suggested that MSCs could be used to dampen immune-mediated diseases and transplant rejection. It is possible that some of the beneficial effects of MSCs might reflect, in part, the trophic and protective activities they exert on injured cells and tissues, rather than resulting from a true transdifferentiation. In immune-mediated diseases, the protective effects might function in concert with the immunosuppressive and anti-inflammatory activities. PMID- 17400511 TI - Mitochondrial redistribution: adding new players to the chemotaxis game. AB - Leukocyte polarization and chemotaxis have a key role in the homeostasis of the immune system and in inflammation. Recent work shows that chemoattractants induce the redistribution of mitochondria towards the uropod of polarized migrating leukocytes through a mechanism involving microtubules and mitochondrial fission. These findings underscore the key role this organelle can have in leukocyte chemotaxis by fuelling motor proteins at their trailing edge. PMID- 17400512 TI - Mast cells--key effector cells in immune responses. AB - Mast cells are best known for their potent effector functions in allergic disorders. In recent years, however, mast cells have been identified to be involved in a surprisingly complex range of immune functions that go far beyond allergies and include the development of autoimmune disorders and peripheral tolerance, and the initiation and maintenance of adaptive and innate host responses. Here, we review the key signals and effector mechanisms that have lately been identified for mast cell functions in these immune responses. PMID- 17400513 TI - Embedding quality improvement and patient safety: the UCLA value analysis experience. AB - Despite pressure from various governmental and non-governmental groups and agencies in the US, many physicians continue to resist the need to measure and improve their clinical practices. Physicians do, however, willingly engage in the process of technology assessment so that new innovations can be introduced into their clinical activities. Technology assessment can be incorporated into a medical staff committee process called value analysis, resulting in both cost savings and cost avoidance. By including a requirement that some approved healthcare technology is monitored for safety and effectiveness within their institutions, members of a medical staff at several academic medical centers within UCLA Healthcare participated eagerly in patient safety and quality improvement programs. PMID- 17400514 TI - Extensive dehiscence of a stentless bioprosthesis with only mild paravalvular regurgitation after replacement of a degenerated bicuspid aortic valve. AB - We report the case of a 54-year-old patient admitted for rehabilitation after implantation of a stentless bioprosthesis due to severe insufficiency of a degenerated bicuspid aortic valve. The patient could fully participate in a standard cardiac rehabilitation program without any signs of haemodynamic instability. Transthoracic echocardiography showed a small mobile subvalvular structure and only mild paravalvular reflux. Transesophageal echocardiography, however, revealed extensive dehiscence of the bioprosthesis with the need for urgent reoperation. We discuss the probable causes and encourage routine echocardiography after valve operations with generous application of transesophageal echocardiography in every suspicious case. PMID- 17400515 TI - Differential expression, induction, and stability of CYP1A4 and CYP1A5 mRNA in chicken and herring gull embryo hepatocytes. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) induces cytochrome P4501A (CYP1A) catalyzed ethoxyresorufin-O-deethylase (EROD) activity in chickens and other avian species. To investigate mechanisms underlying the effectiveness of EROD activity as a biomarker for exposure to dioxin-like compounds in avian models, we characterized inter-species differences in isoform-specific CYP1A mRNA expression, induction, and stability in chickens (Gallus gallus domesticus) and herring gulls (Larus argentatus). Exposure to 100 nM TCDD significantly increased CYP1A4 and CYP1A5 mRNA expression in chicken and herring gull embryo hepatocyte cultures. Chicken CYP1A4 and CYP1A5 were induced 61-fold and 25-fold respectively. The herring gull isoforms were induced 2.2- and 4.3-fold respectively. In both species, the isoform that was preferentially induced exhibited lower constitutive expression. Half-lives of chicken CYP1A4, chicken CYP1A5, and herring gull CYP1A5 mRNA ranged from 5.0 to 7.0 h in cultured hepatocytes. The half-life of herring gull CYP1A4 mRNA was 2.5 h. Our findings indicate that expression, induction, and stability of CYP1A4 and CYP1A5 mRNA are differentially regulated in chickens and herring gulls. In particular, CYP1A4 is preferentially induced in chickens, while CYP1A5 is preferentially induced in herring gulls. We propose that CYP1A5 mRNA expression may be a sensitive biomarker of exposure to dioxin-like compounds in some avian species. PMID- 17400516 TI - Safety and outcomes of laparoscopic gastric bypass surgery in patients 60 years of age and older. AB - BACKGROUND: Many surgeons are hesitant to offer bariatric surgery to patients >60 years of age because of concern of the considerably greater perioperative risk and less weight-control efficacy. We hypothesized that laparoscopic Roux-en-Y gastric bypass (LRYGB) can be performed in this patient population with acceptable morbidity and can achieve effective weight control. METHODS: A retrospective review was performed of patients >60 years of age who had undergone LRYGB at the Bariatric Institute at Cleveland Clinic Florida from 2001 to 2004. The data assessed included age, gender, preoperative and postoperative weight and body mass index (BMI), and postoperative complications. RESULTS: A total of 92 patients >60 years who had undergone LRYGB were reviewed in this study. The mean preoperative weight and BMI was 136.6 kg and 48.4 kg/m(2), respectively. The mean postoperative weight and BMI was 100.3 kg and 35.9 kg/m(2), respectively. The mean percentage of excess weight loss was 53.85%. The early complications were an anastomotic leak in 2 patients (2.2%), intraluminal hemorrhage in 1 patient (1.1%), pulmonary embolus in 1 patient (1.1%), pneumonia in 1 patient (1.1%), and atrial fibrillation in 1 patient (1.1%). The late complications included stenosis at the gastrojejunostomy in 8 patients (8.6%), marginal ulceration in 3 (3.2%), small bowel obstruction in 1 (1.1%), internal hernia in 1 (1.1%), and abdominal wall hernia in 1. No mortality occurred. CONCLUSION: LRYGB can be performed safely and can achieve effective weight control in patients >60 years of age. PMID- 17400517 TI - Bariatric surgery in adolescents: a long-term follow-up study. AB - BACKGROUND: The benefits of bariatric surgery in adult obese patients are well known, but data are lacking regarding the outcome of the surgery in adolescents. The aim of this study was to retrospectively assess the operative morbidity and mortality, percentage of loss of initial excess weight, and the incidence of long term complications and reoperations in a cohort of obese patients who underwent biliopancreatic diversion (BPD) before their 18th birthday. METHODS: A total of 76 adolescent subjects underwent BPD between 1976 and 2005. Of these 78 patients, 7 had Prader-Willi syndrome and 1 had Turner syndrome and were excluded from the study. RESULTS: The patient population comprised 52 girls and 16 boys. Their mean age was 16.8 years, mean body weight at operation was 125 kg (mean body mass index 46 kg/m2). Operative mortality was nil. The mean follow-up was 11 years (range 2-23). The mean percentage of loss of initial excess weight at each patient's longest follow-up was 78%. Before surgery, 33 patients were hypertensive (49%), 11 were dyslipidemic (16%), 3 were hyperglycemic, and 2 had type 2 diabetes. At the longest follow-up period after surgery, only 6 patients were hypertensive, and none were dyslipidemic or diabetic. A total of 19 reoperations were performed in 14 patients (20%), including 7 revisions. Eleven patients developed protein malnutrition 1-10 years after BPD. The long-term mortality rate was 4%. At 4 to 23 years after BPD, 18 of the women had given birth to 28 healthy babies. Three women had had a complicated pregnancy. CONCLUSION: Adolescents can undergo malabsorptive bariatric surgery with excellent long-term weight loss results and an incidence of long-term complications similar to that observed during the 30-year evolution of BPD in our experience. PMID- 17400518 TI - Who is adherent with preoperative psychological treatment recommendations among weight loss surgery candidates? AB - BACKGROUND: Adherence to treatment regimens is critical for success with weight loss surgery. Weight loss surgery patients commonly undergo a psychological evaluation before surgery. When indicated, patients are provided with preoperative behavioral recommendations; yet, little is known about the adherence to these recommendations. In this study, we evaluated the following: (1) level of adherence to behavioral treatment plans; (2) characteristics that differentiate adherent patients from nonadherent patients; and (3) the weight loss of delayed adherent patients compared with those whose surgery was not delayed. METHODS: We performed a case review of psychological evaluations, preoperative behavioral treatment plans, and subsequent adherence rates. RESULTS: Of the 837 patients evaluated, 68 (8%) had significant psychosocial issues and were given behavioral treatment recommendations before surgery. Of these 68 patients, 38 (56%) were adherent and subsequently underwent surgery, and 30 (46%) were nonadherent and were not offered surgery in our program. Adherence did not differ relative to body mass index, age, education, race, marital status, depression, anxiety, or rate of eating disorders. Adherence did differ by gender: 31% percent of the men were adherent to recommendations while 62% of the women were adherent. Individuals with complex treatment plans were less likely to be adherent than those with less complex recommendations. Additionally, increased hostility reduced the adherence rates. Patients adherent to the psychological recommendations had weight loss within 2 years, which was similar to that of patients who did not require behavioral treatment recommendations. CONCLUSION: The results of this study have provided information regarding adherence to preoperative psychological treatment recommendations. Although most patients are psychologically suitable for weight loss surgery at the initial evaluation, a better understanding of the factors influencing the adherence with preoperative behavioral programs could improve care for those with major psychosocial issues. PMID- 17400519 TI - Risk of massive upper gastrointestinal bleeding in gastric bypass patients taking clopidogrel. AB - BACKGROUND: The antiplatelet drug clopidogrel (Plavix) is widely used in patients who have undergone coronary artery stenting or had a stroke. Because morbid obesity is associated with atherosclerosis, some of these patients are candidates for weight loss surgery. We chose to determine the risk of upper gastrointestinal bleeding after gastric bypass in patients taking clopidogrel. METHODS: Patients who took clopidogrel after gastric bypass were identified by specific review of the subset of patients who had had upper gastrointestinal bleeding requiring hospital admission and transfusion. All who bled underwent emergency endoscopy. RESULTS: Of 11 patients taking clopidogrel, 4 (36%) presented with significant upper gastrointestinal bleeding 25-234 days after gastric bypass. All stopped bleeding with discontinuation of the drug and treatment with an intravenous proton pump inhibitor. CONCLUSION: Gastric bypass patients appear to be at high risk of bleeding complications when taking clopidogrel. On the basis of the available published data from another high-risk group (i.e., those with a history of peptic ulcer disease), co-treatment with omeprazole may be indicated when clopidogrel must be continued. PMID- 17400520 TI - Lessons and pitfalls of interactive voice response in medical research. AB - Interactive voice response (IVR) technology is a robust method of data collection that has been underutilized in behavioral medicine and clinical pharmacotherapy research. While it is clear that there is an increasing interest in this technology, published research in the medical field has not described its implementation with sufficient detail to evaluate or replicate the use of interactive voice response systems (IVRS) as part of the methodology. In this article, we hope to shed more light on IVRS as an electronic data capture tool in clinical and behavioral research. We will describe how the IVRS work, the advantages and pitfalls of IVRS, and some of the lessons we have learned from administering a number of clinical studies using IVRS. Readers will gain a better understanding of this technology, which will enable them to optimize its usage in clinical research. PMID- 17400521 TI - Population genetic aspects and phenotypic plasticity of ventilatory responses in high altitude natives. AB - Highland natives show unique breathing patterns and ventilatory responses at altitude, both at rest and during exercise. For many ventilatory traits, there is also significant variation between highland native groups, including indigenous populations in the Andes and Himalaya, and more recent altitude arrivals in places like Colorado. This review summarizes the literature in this area with some focus on partitioning putative population genetic differences from differences acquired through lifelong exposure to hypoxia. Current studies suggest that Tibetans have high resting ventilation (V (E)), and a high hypoxic ventilatory response (HVR), similar to altitude acclimatized lowlanders. Andeans, in contrast, show low resting V (E) and a low or "blunted" HVR, with little evidence that these traits are acquired via lifelong exposure. Resting V (E) of non-indigenous altitude natives is not well documented, but lifelong hypoxic exposure almost certainly blunts HVR in these groups through decreased chemosensitivity to hypoxia in a process known as hypoxic desensitization (HD). Together, these studies suggest that the time course of ventilatory response, and in particular the origin or absence of HD, depends on population genetic background i.e., the allele or haplotype frequencies that characterize a particular population. During exercise, altitude natives have lower V (E) compared to acclimatized lowland controls. Altitude natives also have smaller alveolar-arterial partial pressure differences P(AO2) - P(aO2) during exercise suggesting differences in gas exchange efficiency. Small P(AO2) - P(aO2) in highland natives of Colorado underscores the likely importance of developmental adaptation to hypoxia affecting structural/functional aspects of gas exchange with resultant changes in breathing pattern. However, in Andeans, at least, there is also evidence that low exercise V (E) is determined by genetic background affecting ventilatory control independent of gas exchange. Additional studies are needed to elucidate the effects of gene, environment, and gene-environment interaction on these traits, and these effects are likely to differ widely between altitude native populations. PMID- 17400522 TI - HPLC method for determination of in vitro delivery through and into porcine skin of adefovir (PMEA). AB - A simple HPLC/UV method for the determination of the transdermal permeation and dermal penetration of a broad-spectrum antiviral drug adefovir (PMEA) was developed. The separation was achieved on a C18 column with the mobile phase composed of 10 mM KH2PO4 and 2 mM Bu4NHSO4 at pH 6.0 and 7% acetonitrile. The method was validated with respect to selectivity, linearity (0.1-50 microg/ml), precision, accuracy, and stability. Transdermal permeation of 2% PMEA was studied in vitro using the Franz diffusion cell and porcine skin. The flux values were 1.8, 3.0, and 0.6 microg/cm2/h from aqueous donor samples at pH 3.4 and 7.4, and isopropyl myristate, respectively. The respective skin concentrations at 48 h were 294, 263, and 971 microg/g from these vehicles. These results will serve as a lead for further studies on transdermal and topical delivery of antivirals from the group of acyclic nucleoside phosphonates. PMID- 17400523 TI - The suitability of DEAE-Cl active groups on customized poly(GMA-co-EDMA) continuous stationary phase for fast enzyme-free isolation of plasmid DNA. AB - The creation of a commercially viable and a large-scale purification process for plasmid DNA (pDNA) production requires a whole-systems continuous or semi continuous purification strategy employing optimised stationary adsorption phase(s) without the use of expensive and toxic chemicals, avian/bovine-derived enzymes and several built-in unit processes, thus affecting overall plasmid recovery, processing time and economics. Continuous stationary phases are known to offer fast separation due to their large pore diameter making large molecule pDNA easily accessible with limited mass transfer resistance even at high flow rates. A monolithic stationary sorbent was synthesised via free radical liquid porogenic polymerisation of ethylene glycol dimethacrylate (EDMA) and glycidyl methacrylate (GMA) with surface and pore characteristics tailored specifically for plasmid binding, retention and elution. The polymer was functionalised with an amine active group for anion-exchange purification of pDNA from cleared lysate obtained from E. coli DH5alpha-pUC19 pellets in RNase/protease-free process. Characterization of the resin showed a unique porous material with 70% of the pores sizes above 300 nm. The final product isolated from anion-exchange purification in only 5 min was pure and homogenous supercoiled pDNA with no gDNA, RNA and protein contamination as confirmed with DNA electrophoresis, restriction analysis and SDS page. The resin showed a maximum binding capacity of 15.2 mg/mL and this capacity persisted after several applications of the resin. This technique is cGMP compatible and commercially viable for rapid isolation of pDNA. PMID- 17400524 TI - Use of magnetic poly(glycidyl methacrylate) monosize beads for the purification of lysozyme in batch system. AB - The hydrophobic affinity ligand L-tryptophan immobilized magnetic poly(glycidyl methacrylate) [m-poly(GMA)] beads in monosize form (1.6 microm in diameter) were used for the affinity purification of lysozyme from chicken egg white. The m poly(GMA) beads were prepared by dispersion polymerization in the presence of Fe3O4 nano-powder. The epoxy groups of the m-poly(GMA) beads were converted into amino groups with 1,6 diaminohexane (i.e., spacer arm). l-tryptophan was then covalently immobilized on spacer arm attached m-poly(GMA) beads. Elemental analysis of immobilised L-tryptophan for nitrogen was estimated as 42.5 micromol/g polymer. Adsorption studies were performed under different conditions in a batch system (i.e., medium pH, protein concentration and temperature). Maximum lysozyme adsorption amount of m-poly(GMA) and m-poly(GMA)-L-tryptophan beads were 1.78 and 259.6 mg/g, respectively. The applicability of two kinetic models including pseudo-first order and pseudo-second order model was estimated on the basis of comparative analysis of the corresponding rate parameters, equilibrium adsorption capacity and correlation coefficients. Results suggest that chemisorption processes could be the rate-limiting step in the adsorption process. It was observed that after 10 adsorption-elution cycle, m-poly(GMA)-L tryptophan beads can be used without significant loss in lysozyme adsorption capacity. Purification of lysozyme from egg white was also investigated. Purification of lysozyme was monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. It was found to be successful in achieving purification of lysozyme in a high yield of 76% with a purification fold of 71 in a single step. The specific activity of the eluted lysozyme (62,580 U/mg) was higher than that obtained with a commercially available pure lysozyme (Sigma (60,000 U/mg). PMID- 17400525 TI - Development and validation of an LC/MS/MS assay for mycophenolic acid in human peripheral blood mononuclear cells. AB - The aim was to develop a LC/MS/MS method able to quantify mycophenolic acid (MPA) in the peripheral blood mononuclear cells (PBMCs) of transplanted patients. PBMCs were isolated from blood by a density gradient separation. The chromatographic separation was carried out on a Zorbax Stable Bond CN, 150 mmx2.1 mm, and MS/MS detection was performed after positive electrospray ionisation of the protonated parent ion. The calibration range was from 0.25 to 100 ng/sample. Extraction from the cells and ionisation recoveries reached 73.5 and 37.9%, respectively. Inaccuracy was always <10% with CVs<15%. MPA was stable at room temperature in the autosampler over 48 h and at -20 degrees C over 1.5 months. Application to clinical samples taken from patients treated with mycophenolate mofetil indicated that the method is suitable for measuring intracellular MPA. PMID- 17400526 TI - Complex retention behavior of pyrimidines on biomembrane-mimic immobilized artificial-membrane phase. AB - The influence of the chemical substitutions on the interfacial interactions of pyrimidines with the phospholipid-mimic immobilized-artificial-membrane (IAM) chromatographic stationary phase was evaluated. Monocyclic pyrimidine nucleic acid bases (nucleobases) were revealed behaving differently from their bicyclic purine counterparts substantially. The computed electrostatic potential surfaces for both the IAM phase and the interacting nucleobases are intuitive in deconvoluting the retention patterns of pyrimidines molecularly. A structure retention model has also been derived using quantitative 3D-QSAR methodology pertinent to the IAM-retention of pyrimidines for the potential use in molecular design. IAM phase is found particularly suitable in assessing the retention of pyrimidines with bulky or elongated exocyclic substituents in the biological context than the alkyl-based chromatographic counterparts. PMID- 17400527 TI - Oral bioavailability of curcumin in rat and the herbal analysis from Curcuma longa by LC-MS/MS. AB - This study presents a validated liquid chromatography technique coupled with tandem mass spectrometry (LC-MS/MS) to measure curcumin in rat plasma and provide curcuminoids analysis from the extract of Curcumin longa L. This method was applied to investigate the pharmacokinetics of curcumin in a freely moving rat. The analytes were separated by a reversed phase C18 column (150x4.6 mm I.D., particle size 5 microm) and eluted with acetonitrile-1mM HCOOH mobile phase (70:30, v/v) with a flow rate of 0.8 ml/min in rat plasma and herbal extracts. Multiple reaction monitoring (MRM) was used to monitor the transition of the deprotonated molecule m/z of 367 [M-H]- to the product ion 217 for curcumin, a m/z of 337-217 for demethoxycurcumin and a m/z of 265-224 for honokiol (internal standard) analysis. The limit of detection (LOD) and quantification (LOQ) of curcumin in the rat plasma were 1 and 5 ng/ml, respectively. The method was linear in the range of 5-1000 ng/ml with a coefficient of correlation greater than 0.996 in the rat plasma. After curcumin (500 mg/kg, p.o.) administration, the maximum concentration (Cmax) and the time to reach maximum concentration (Tmax) were 0.06+/-0.01 microg/ml and 41.7+/-5.4 min, respectively. The elimination half-life (t1/2,beta) were 28.1+/-5.6 and 44.5+/-7.5 min for curcumin (500 mg/kg, p.o.) and curcumin (10 mg/kg, i.v.), respectively. The oral bioavailability was about 1%. PMID- 17400528 TI - Symptomatic gastric heterotopic pancreas: clinical presentation and review of the literature. AB - Heterotopic pancreas is usually asymptomatic. Associated complications include ulceration, bleeding and obstruction, especially gastric outlet obstruction. Acute and chronic pancreatitis including malignant change have been reported. These lesions are often difficult to differentiate clinically from other gastrointestinal diseases including primary gastric malignancy. Histological examination is mandatory to make a correct diagnosis. Limited surgical excision has been shown to be safe and adequate for heterotopic pancreas. We present a patient with heterotopic pancreas who presented with a non-healing gastric ulcer. PMID- 17400529 TI - Eliciting and storing operative information from expert surgeons using surgical scripts: a content analysis study. AB - INTRODUCTION: Surgical trainees learn many of the minutiae of operative technique at the operating table. As trainees' operating exposure decreases, so will the opportunity for the trainee to work with and learn from a range of different surgeons. Surgical scripts provide a complementary source of expert operative information from individual surgeons. This pilot study examines whether a surgical script is an effective way of eliciting the combined operative information from a group of surgical experts. METHODS: One operative step was taken from a surgical script: tracing the right ureter in a right hemicolectomy. A group of surgeons were asked to review the information supporting this step in the script and to add all their own hints and tips for the step. The comments elicited from the surgeons were subjected to content analysis. RESULTS: The original script contained 15 points of information backing up the ureter step, more than 3 times as many as in the most detailed operative textbook found. Nineteen surgeons contributed 50 additional comments, providing more detail, extra information, alternative techniques and different opinions. Some of the comments have been incorporated into a revised script. CONCLUSION: This pilot study showed that a surgical script is an effective way of eliciting and storing detailed operative information from a group of surgeons. The relevance of these findings is discussed. The resulting scripts on line should be helpful in optimising trainees' operating time. PMID- 17400530 TI - Innervation of three weight-bearing areas of the foot: an anatomic study and clinical implications. AB - The aim of this cadaver study is to improve our knowledge on the anatomy of the sensory fibres of the three weight-bearing areas of the plantar region. Previous studies mainly focused on the innervation of the heel but the innervation of the other two weight-bearing areas over the most medial and lateral metatarses have been neglected and are not well known. The study was carried out on 10 feet of five male cadavers. The tibial nerve was dissected down to the fat pads over the heel and the first and fifth metatarsal heads under the microscope. The distances of the branching point of the tibial nerve and origins of the medial and inferior calcaneal nerves to a line drawn from the centre of the medial malleolus to the centre of the calcaneous were all measured. The tibial nerve was divided into two branches called the lateral and medial plantar nerves 23.45 mm proximal to the predefined axis. The medial plantar nerve passed underneath the abductor hallucis muscle and gave two sensory branches to the fat pad over the first metatarsal head. The lateral plantar nerve coursed beneath the abductor hallucis and flexor digitorum brevis muscles and supplied innervation of the fat pad over the fifth metatarsal head. The sensory innervation of the heel was provided by medial calcaneal and inferior calcaneal nerves. The medial calcaneal nerve originated from the tibial nerve 41.89 mm proximal to the axis. It divided into two or three branches innervating the fat pad over the heel. The inferior calcaneal nerve originated from the lateral plantar nerve (70%) or the medial calcaneal nerve (30%) 10.66 mm proximal to the axis. This study describes the sensory fibres to the heel and the previously neglected weight-bearing areas over the first and fifth metatarses. Reconstruction of defects in these areas is very difficult so every attempt should be made to protect the sensory fibres during any surgical procedure. PMID- 17400531 TI - Skeletonised retrograde distal perforator island fasciocutaneous flaps for leg and foot defects. AB - Defects involving the distal leg and foot are frequently encountered following various aetiological factors. Paucity of local tissue causes surgeons to resort to the retrograde peninsular flap, the cross leg flap or the free flap. With specific knowledge of perforators, the fasciocutaneous flap from the calf area can be transferred to the defect in a single stage based on skeletonised distal perforators. The surgical anatomy, flap planning and procedure have been detailed. Nineteen patients were treated during the period 1995 to 2005. The perforators were identified preoperatively by audio Doppler. The flaps were marked and dissected proximal to the defect skeletonising the distal perforators under loupe magnification and transferred to the defect in a single stage. The donor site was skin grafted. Out of 19 cases, 16 flaps healed uneventfully, one flap necrosed completely and in two cases there was marginal necrosis. The cases were followed up for 2-10 years with an average of 6 years. With detailed knowledge of perforators one can safely reconstruct distal moderate-size defects of the lower limb in a single stage, thus having the benefits of free tissue transfer without resorting to microsurgery. This technique has proved to be an advancement in the reconstructive repertoire allowing flaps of non conventional dimensions to be perfused by skeletonised perforators. PMID- 17400532 TI - Molecular study of six families originating from the Middle-East and presenting with autosomal recessive osteopetrosis. AB - Autosomal recessive osteopetrosis is a severe hereditary bone disease whose cellular basis is in the osteoclast, but with heterogeneous molecular defects. We hereby report the clinical and the molecular study of seven patients affected by the recessive form of osteopetrosis (ARO) from six families originating from the Middle-East: four from Lebanon and two from Syria. Parental consanguinity was found in five families. The mean age of diagnosis was 3 months. Failure to thrive, prominent forehead, exophthalmia, optic atrophy, hepatosplenomegaly, neurological manifestations, anaemia, thrombocytopenia, hypocalcaemia, elevated hepatic enzymes and acid phosphatase, and an early fatal outcome were common. Macrocephaly, strabismus, and brain malformations were relatively less common. Mutations were identified in two genes: TCIRG1 and OSTM1. Phenotype-genotype correlation is discussed. PMID- 17400533 TI - Reactivation of tuberculosis by tumor necrosis factor neutralization. AB - Tumor necrosis factor (TNF) is required in the control of infection with Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis. TNF is essential and non-redundant for forming microbiocidal granulomas, and cannot be replaced by other members of the TNF family. We established a model of latent Mtb infection in mice, allowing investigation of the reactivation of latent Mtb as observed in patients receiving TNF-neutralizing therapy used in rheumatoid arthritis and Crohn's disease. Antibody neutralization of TNF is able to reactivate clinically silent Mtb infection. Using mutant mice expressing solely membrane, but not soluble TNF, we demonstrated that membrane TNF is sufficient to control acute Mtb infection. Therefore, we hypothesize that TNF-neutralizing therapy, sparing membrane TNF, may have an advantage as compared to complete neutralization. In conclusion, endogenous TNF is critical for the control of tuberculosis infection. Genetic absence or pharmacological neutralization of TNF results in uncontrolled infection, while selective neutralization might retain the desired anti-inflammatory effect but reduce the infectious risk. PMID- 17400534 TI - Continuous large-scale production of the cytokine CXCL8 from a novel porcine cell line. AB - Cytokine production from two unstimulated porcine cell lines (SL-24 and SK-L) was examined using porcine cytokine detection ELISA kits and RT-PCR. Porcine IL-1 alpha, IL-6, and CXCL8 were detected in all samples examined. In particular, the SL-24 cell line (derived from bone marrow cells of a malignant lymphoma-affected pig), produced large amounts of porcine CXCL8. Flow cytometer analysis showed the cell line to be strongly CD44 positive, and was therefore considered to be of monocyte or macrophage origin. Porcine CXCL8 production was greatest (83.86 +/- 32.33 ng/mL) at six days post-cultivation. The SK-L cell line (derived from porcine kidney) also produced CXCL8, but production was less than 1.5 ng/mL. Porcine CXCL8 from the SL-24 cell line, induced chemotactic activity in porcine neutrophils, while the production of CXCL8 from the SL-24 cell line was inhibited by dexamethasone, which suggests that the mechanism of CXCL8 production is related to an NF-kappaB binding site. The production of CXCL8 from the SL-24 cell line was enhanced by the addition of recombinant porcine IL-15, which is the first reported observation of such CXCL8 production. Cloning of the SL-24 cell line by limited dilution revealed two types of cells present in the starting population. One cell type, designated as long-form cells (LC), produced large amounts of CXCL8, while the other, designated short-form cells (SC), produced small amounts of the cytokine. The LC cells were adapted to grow in serum-free medium in which they produced large amounts of CXCL8. The large-scale production of porcine CXCL8 from the SL-24 cell line will be of value in determining the mechanism of cytokine production and as a source of naturally produced porcine CXCL8. PMID- 17400535 TI - Polyvalent 23 epitope polysaccharide pneumonia vaccine induced effective protection through strain-adapted effector mechanisms as demonstrated by the different cytokine responses in mice challenged with two different strains of Streptococcus pneumoniae. AB - We used a Balb/c mouse model of pneumococcal pneumonia to investigate the protection mechanisms induced by immunization with a polyvalent 23 epitope polysaccharide pneumonia vaccine. Groups of mice were injected x 4 times s.c. within one month, with this vaccine preparation. Mice were subsequently challenged at day 45, with a lethal, intratracheal inoculum of two strains of Streptococcus pneumoniae - either a highly virulent and strongly immunogenic serotype 3 strain (P4241), or a less virulent and weakly immunogenic serotype 19F strain (P15986). The intratracheal S. pneumoniae challenge-induced lethality, antibody response, bacterial clearance, and cytokine secretions were monitored to analyze the strain-adapted effector mechanisms. Pulmonary levels of TNFalpha, IL 6, IL-1 beta, MIP-1 alpha, KC, MCP-1/JE and MIP-2 cytokines were determined up to 48 hours post-infection. Survival rates were 82% and 100% among vaccinated animals challenged at day 45 with P4241, and P1598 mice respectively, and 0% in non-vaccinated mice (p<0.001). Survival was associated with a rapid bacterial clearance from blood and lungs, which similar for the two strains. Immunization induced a serotype-specific antibody response. Kinetics of the cytokine profile in the lung following intratracheal inoculation with the 4241 strain was different in animals vaccinated 45 days previously, compared to naive, control mice. Generally speaking the bacterial-induced inflammatory cytokine response induced with the 4241 strain was much weaker in vaccinated animals than in control mice. The only cytokines showing a greater increase in vaccinated mice compare to control animals were IL-1 beta, KC and MCP-1. Production of TNFalpha and IL-6 was lower in vaccinated animals than in controls. At variance with the previous bacteria strain-induced cytokine profile, infection with the P15986 strain induced a strong inflammatory response, with a substantial increase in all the cytokine tested, which was similar in vaccinated and in naive, control animals, except for MIP-1 alpha, which was the only mediator significantly more produced by vaccinated animals than by naive, control mice following P15986 infection. The distinct cytokine profiles, which were observed in this study depending upon the two strains of S. pneumoniae used for challenge, demonstrated that protection against each strain was obtained through a different defence strategy. PMID- 17400536 TI - Relevance of transforming growth factor-beta1, interleukin-8, and tumor necrosis factor-alpha polymorphisms in patients with chronic pancreatitis. AB - Cytokine regulation may be an important factor in the susceptibility for the development of chronic pancreatitis; transforming growth factor-beta1 (TGF-beta1) plays a central role in the pathogenesis of pancreatic fibrogenesis. The aim of our study was to analyse the relevance of TGF-beta1, interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha) polymorphisms in patients with chronic pancreatitis. PATIENTS: of the 83 patients enrolled in the study, 43 were treated medically and 40 patients underwent surgical intervention. Healthy blood donors (n=75) served as controls. METHODS: the polymorphisms of TGF-beta1 +869 T--> C and IL-8 -251 T-->A were determined by the ARMS method, while that of TNF-alpha 308 was investigated using NcoI RFLP. RESULTS: there was a higher frequency (50%) of the TT genotype of TGF-beta1 +869, with a concomitantly higher TGF-beta1 level in the plasma (5.2 +/- 1.7 ng/mL) of patients with chronic pancreatitis than in healthy blood donors (28% and 2.8 +/- 0.9 ng/mL respectively). The number of TT homozygotes differed significantly between the patients who underwent surgical intervention and the controls, and even between the surgical and the non-surgical patients. The frequency of the T/A genotype with higher IL-8 production, was significantly higher in both groups of patients than in the controls (58% and 58% versus 40%). No correlation was found between the TNF-alpha -308 polymorphism and chronic pancreatitis. CONCLUSIONS: correlations of the TGF-beta1 and IL-8 single nucleotide polymorphisms (SNPs) with chronic pancreatitis underline the importance of these cytokines in the pathomechanism of the disease. Moreover, it seems that the TT genotype of +869 TGF-beta1 might be a risk factor for the development of a severe form of chronic pancreatitis, and could serve as a prognostic sign for any future surgical intervention or even repeat surgery. Further studies on a larger group of patients, in addition to a follow-up study, are necessary to confirm this preliminary observation. PMID- 17400537 TI - Both viable and killed Candida albicans cells induce in vitro production of TNF alpha and IFN-gamma in murine cells through a TLR2-dependent signalling. AB - The in vitro production of TNF-alpha and IFN-gamma in response to Candida albicans was investigated in wild type, TLR2-/- and TLR4-/- murine cells. TLR2-/- resident peritoneal macrophages showed a strong impairment of TNF-alpha production in response to viable and non-viable (heat-killed, antimycotic-treated and formaldehyde-fixed) yeasts and hyphae (germ tube-bearing cells) of the high virulence C. albicans ATCC 26555 strain, as compared with macrophages from wild type and TLR4-/- mice. The in vitro production of IFN-gamma was investigated in murine splenocytes obtained three days after intravenous injection with the low virulence, non-germinative C. albicans PCA2 strain, and again, TLR2-/- splenocytes showed a strong impairment of the in vitro production of IFN-gamma in response to non-viable (heat-killed, antimycotic-treated and formaldehyde-fixed) C. albicans ATCC 26555 yeasts, as compared with splenocytes of TLR4-/- and wild type mice. These results indicate that the TLR2-mediated recognition of C. albicans leading to a proinflammatory Th1 host response appears to be well conserved in killed C. albicans cells, regardless of the inactivating treatment employed. PMID- 17400538 TI - From cognitive to neural models of working memory. AB - Working memory refers to the temporary retention of information that was just experienced or just retrieved from long-term memory but no longer exists in the external environment. These internal representations are short-lived, but can be stored for longer periods of time through active maintenance or rehearsal strategies, and can be subjected to various operations that manipulate the information in such a way that makes it useful for goal-directed behaviour. Empirical studies of working memory using neuroscientific techniques, such as neuronal recordings in monkeys or functional neuroimaging in humans, have advanced our knowledge of the underlying neural mechanisms of working memory. This rich dataset can be reconciled with behavioural findings derived from investigating the cognitive mechanisms underlying working memory. In this paper, I review the progress that has been made towards this effort by illustrating how investigations of the neural mechanisms underlying working memory can be influenced by cognitive models and, in turn, how cognitive models can be shaped and modified by neuroscientific data. One conclusion that arises from this research is that working memory can be viewed as neither a unitary nor a dedicated system. A network of brain regions, including the prefrontal cortex (PFC), is critical for the active maintenance of internal representations that are necessary for goal-directed behaviour. Thus, working memory is not localized to a single brain region but probably is an emergent property of the functional interactions between the PFC and the rest of the brain. PMID- 17400539 TI - The reign of typicality in semantic memory. AB - This paper begins with a brief description of a theoretical framework for semantic memory, in which processing is inherently sensitive to the varying typicality of its representations. The approach is then elaborated with particular regard to evidence from semantic dementia, a disorder resulting in relatively selective deterioration of conceptual knowledge, in which cognitive performance reveals ubiquitous effects of typicality. This applies to frankly semantic tasks (like object naming), where typicality can be gauged by the extent to which an object or concept is characterized by shared features in its category. It also applies in tasks apparently requiring only access to a 'surface' representation (such as lexical decision) or translation from one surface representation to another (like reading words aloud), where typicality is defined in terms of the structure of the surface domain(s). The effects of surface-domain typicality also appear early in the time course of word and object processing by normal participants, as revealed in event-related potential studies. These results suggest that perceptual and conceptual processing form an interactive continuum rather than distinct stages, and that typicality effects reign throughout this continuum. PMID- 17400540 TI - Cloning, expression, purification and characterization of fructose-1,6 bisphosphate aldolase from Anoxybacillus gonensis G2. AB - The fructose-1,6-bisphosphate aldolase gene from the thermophilic bacterium, Anoxybacillus gonensis G2, was cloned and sequenced. Nucleotide sequence analysis revealed an open reading frame coding for a 30.9 kDa protein of 286 amino acids. The amino acid sequence shared approximately 80-90% similarity to the Bacillus sp. class II aldolases. The motifs that are responsible for the binding of a divalent metal ion and catalytic activity completely conserved. The gene encoding aldolase was overexpressed under T7 promoter control in Escherichia coli and the recombinant protein purified by nickel affinity chromatography. Kinetic characterization of the enzyme was performed at 60 degrees C, and K(m) and V(max) were found to be 576 microM and 2.4 microM min(-1) mg protein(-1), respectively. Enzyme exhibits maximal activity at pH 8.5. The activity of enzyme was completely inhibited by EDTA. PMID- 17400541 TI - Head injury resulting from scooter accidents in Rome: differences before and after implementing a universal helmet law. AB - OBJECTIVES: To estimate the incidence rates and related determinants of head injuries before and after the implementation of a new universal helmet law in Italy. METHODS: The investigation took place in the Emergency Room of the Accident and Emergency Department, Teaching Hospital 'Umberto I', Rome, in 1999 (before the new law), and 2000 (two periods after the new law). Personal data, injury circumstances, helmet use, others involved in injury and health data, were collected. Incidence rates by time period were calculated and differences between groups were analysed. Logistic regression models were conducted to assess the association between head trauma and potential determinants. RESULTS: The incidence rate of all injury among scooter riders rose from 64.36/10,000 person years before the new law (1999) to 98.05/10,000 person years afterwards. The incidence rate of head trauma among scooter riders showed an opposite trend, i.e. a decrease from 26.65/10,000 person-years (1999) to 8.88/10,000 person-years in the second post-legislation period. Helmet use among injured scooter riders rose from 5% before the new law to 90% afterwards. Helmet use is a protective factor for being a patient with head trauma, in all the periods considered (OR 0.24 0.44). After implementation of the new law, age (18+) showed a protective effect as well (OR 0.42-0.44). CONCLUSION: Helmet use has a protective effect on head trauma among scooter riders. One year after implementing a universal law, helmet use has increased substantially and a sharp reduction in head trauma among persons older than 18 years could be observed. PMID- 17400542 TI - Prevalence and factors related to malocclusion and orthodontic treatment need in children and adolescents in Italy. AB - BACKGROUND: The purpose of this study was to determine prevalence of malocclusions, normative and perceived orthodontic treatment need and related risk factors in schoolchildren in Italy. DESIGN: A random sample of 1000 11-15 years old children was selected from randomly selected schools in Catanzaro (Italy). Parents completed a questionnaire on sociodemographic, orthodontic history and perception of their child orthodontic treatment need. Children were interviewed on utilization of dental services, perception of orthodontic treatment need and use of orthodontic devices. The Decayed, Missing, Filled Teeth (DMFT) index and the Dental Health Component (DHC) of the Index of Orthodontic Treatment Need (IOTN) were assessed. The Aesthetic Component (AC) of IOTN was assessed by parents, children and dentist. RESULTS: A total of 546 children participated in the study. Three hundred and twenty-five subjects (59.5%) were assigned a 4 or 5 IOTN score, thus requiring orthodontic treatment. Definite treatment need (AC score 8-10) was reported for 8.6% of subjects by the orthodontist, 5.4% by parents and 3.2% by children. Higher DMFT significantly predicted orthodontic treatment need according to IOTN. Perceived orthodontic treatment need was significantly predicted by orthodontic treatment need for crowding/spacing and overjet. CONCLUSION: A high proportion of children needs normative orthodontic treatment and perceptions of orthodontic treatment do not overlap with normative need. PMID- 17400543 TI - Obesity in asylum seekers' children in The Netherlands--the use of national reference charts. AB - BACKGROUND: Growth assessment can be used to monitor health at individual and population level. For asylum seekers' children with different geographic backgrounds, growth reference values are frequently not available. We assessed nutritional condition and growth of asylum seekers' children upon arrival and follow-up in The Netherlands, using national growth charts, and related these parameters to geographic origin. METHODS: Height and weight of 135 children originating from Africa (n=47), Central Asia (n=41), and Eastern Europe (n=47), were assessed longitudinally (median follow-up 3 years, range 1-8 years). Body mass-index (BMI) was calculated, and overweight and obesity were defined according the international BMI cut-off values for age and gender. RESULTS: Upon arrival at a median age of 4.5 years (range 0-11.5 years), 13% of the children were small for age (below -2 SD of the Dutch height for age reference), which decreased to 5% during follow-up (P<0.05). During follow-up, 90% of the height measurements in boys and 85% in girls were within the normal range (+/-2 SD) of the Dutch references. The proportion of children with overweight including obesity increased from 15% at arrival to 21% during follow-up (P<0.05). Irrespective of age, children originating from Africa were taller than children from Central Asia or Eastern Europe at follow up (P<0.05). Overweight and obesity was most prominent among children of Eastern Europe. CONCLUSION: Dutch national reference values allow monitoring growth and the development of overweight or obesity in asylum seekers' children in The Netherlands. Prevention strategies to reduce the development of overweight and obesity among these children seem warranted. PMID- 17400544 TI - Cell phenotype-specific down-regulation of Smad3 involves decreased gene activation as well as protein degradation. AB - Signaling by transforming growth factor-beta (TGF-beta), a regulator of several biological processes, including renal fibrosis, is mediated, in part, by the Smad proteins. Tight control of Smad level and activity is critical for proper TGF beta biological functions. Here, we have investigated the mechanisms involved in regulating Smad3 expression. In human glomerular mesangial cells, Smad3 protein levels were specifically reduced by 24 h of TGF-beta1 treatment, whereas Smad2 and Smad4 levels were not. TGF-beta1 increased endogenous Smad3 ubiquitination, and proteasome inhibitor treatment blocked TGF-beta1-mediated Smad3 down regulation resulting in accumulation of ubiquitinated Smad3. These data support the concept that Smad3 down-regulation occurs via degradation by the ubiquitin/proteasome machinery. However, changes in Smad3 protein levels were also paralleled by changes in Smad3 mRNA expression. TGF-beta1 did not decrease Smad3 mRNA stability, but it significantly inhibited Smad3 promoter activity. In renal tubular epithelial cells, decreased Smad3 levels were observed only after exposure to TGF-beta1 for longer time periods (5-7 days) that paralleled epithelial-to-mesenchymal transition, as determined by increased expression of smooth muscle alpha-actin and decreased expression of E-cadherin. Decline in Smad3 expression also occurred in kidneys after unilateral ureteral obstruction, a model of tubulointerstitial fibrosis associated with TGF-beta up-regulation and epithelial-to-mesenchymal transition. Our data show for the first time that TGF beta1 modulates the expression of a receptor-activated Smad at both the protein and transcriptional level. Smad3 down-regulation could represent a feedback loop controlling TGF-beta signaling in a cell phenotype-specific manner. PMID- 17400545 TI - R-spondin1 is a high affinity ligand for LRP6 and induces LRP6 phosphorylation and beta-catenin signaling. AB - R-spondin proteins are newly identified secreted molecules that activate beta catenin signaling. However, the mechanism of R-spondin action and its relationship with Wnt signaling remain unclear. Here we show that human R spondin1 (hRspo1) is a high affinity ligand for the Wnt co-receptor LRP6 (K(d) = 1.2 nm). hRspo1 induces glycogen synthase kinase 3-dependent phosphorylation and activation of LRP6. DKK1, an LRP6 antagonist, inhibits hRspo1-induced LRP6 phosphorylation. We further demonstrate that hRspo1 synergizes with Frizzled5 in Xenopus axis induction assays and induces the phosphorylation of Dishevelled, a cytoplasmic component downstream of Frizzled function. Our study reveals interesting similarity and distinction between Wnt and R-spondin signaling. PMID- 17400546 TI - Brorin, a novel secreted bone morphogenetic protein antagonist, promotes neurogenesis in mouse neural precursor cells. AB - We identified a gene encoding a novel secreted protein in mice and humans and named it Brorin. Mouse Brorin consists of 324 amino acids with a putative secreted signal sequence at its amino terminus and two cysteine-rich domains in its core region. Positions of 10 cysteine residues in the domains of Brorin are similar to those in the cysteine-rich domains of members of the Chordin family. However, the amino acid sequence of Brorin is not significantly similar to that of any other member of the Chordin family, indicating that Brorin is a unique member of the family. Mouse Brorin protein produced in cultured cells was efficiently secreted into the culture medium. The protein inhibited the activity of bone morphogenetic protein 2 (BMP2) and BMP6 in mouse preosteoblastic MC3T3-E1 cells. Mouse Brorin was predominantly expressed in neural tissues in embryos and also predominantly expressed in the adult brain. In the brain, the expression was detected in neurons, but not glial cells. The neural tissue-specific expression profile of Brorin is quite distinct from that of any other member of the Chordin family. Brorin protein promoted neurogenesis, but not astrogenesis, in mouse neural precursor cells. The present findings indicate that Brorin is a novel secreted BMP antagonist that potentially plays roles in neural development and functions. PMID- 17400547 TI - Synapsin phosphorylation by SRC tyrosine kinase enhances SRC activity in synaptic vesicles. AB - Synapsins are synaptic vesicle-associated phosphoproteins implicated in the regulation of neurotransmitter release. Synapsin I is the major binding protein for the SH3 domain of the kinase c-Src in synaptic vesicles. Its binding leads to stimulation of synaptic vesicle-associated c-Src activity. We investigated the mechanism and role of Src activation by synapsins on synaptic vesicles. We found that synapsin is tyrosine phosphorylated by c-Src in vitro and on intact synaptic vesicles independently of its phosphorylation state on serine. Mass spectrometry revealed a single major phosphorylation site at Tyr(301), which is highly conserved in all synapsin isoforms and orthologues. Synapsin tyrosine phosphorylation triggered its binding to the SH2 domains of Src or Fyn. However, synapsin selectively activated and was phosphorylated by Src, consistent with the specific enrichment of c-Src in synaptic vesicles over Fyn or n-Src. The activity of Src on synaptic vesicles was controlled by the amount of vesicle-associated synapsin, which is in turn dependent on synapsin serine phosphorylation. Synaptic vesicles depleted of synapsin in vitro or derived from synapsin null mice exhibited greatly reduced Src activity and tyrosine phosphorylation of other synaptic vesicle proteins. Disruption of the Src-synapsin interaction by internalization of either the Src SH3 or SH2 domains into synaptosomes decreased synapsin tyrosine phosphorylation and concomitantly increased neurotransmitter release in response to Ca(2+)-ionophores. We conclude that synapsin is an endogenous substrate and activator of synaptic vesicle-associated c-Src and that regulation of Src activity on synaptic vesicles participates in the regulation of neurotransmitter release by synapsin. PMID- 17400548 TI - Stringent 3Q.1R composition of the SNARE 0-layer can be bypassed for fusion by compensatory SNARE mutation or by lipid bilayer modification. AB - SNARE proteins form bundles of four alpha-helical SNARE domains with conserved polar amino acids, 3Q and 1R, at the "0-layer" of the bundle. Previous studies have confirmed the importance of 3Q.1R for fusion but have not shown whether it regulates SNARE complex assembly or the downstream functions of assembled SNAREs. Yeast vacuole fusion requires regulatory lipids (ergosterol, phosphoinositides, and diacylglycerol), the Rab Ypt7p, the Rab-effector complex HOPS, and 4 SNAREs: the Q-SNAREs Vti1p, Vam3p, and Vam7p and the R-SNARE Nyv1p. We now report that alterations in the 0-layer Gln or Arg residues of Vam7p or Nyv1p, respectively, strongly inhibit fusion. Vacuoles with wild-type Nyv1p show exquisite discrimination for the wild-type Vam7p over Vam7(Q283R), yet Vam7(Q283R) is preferred by vacuoles with Nyv1(R191Q). Rotation of the position of the arginine in the 0-layer increases the K(m) for Vam7p but does not affect the maximal rate of fusion. Vam7(Q283R) forms stable 2Q.2R complexes that do not promote fusion. However, fusion is restored by the lipophilic amphiphile chlorpromazine or by the phospholipase C inhibitor U73122, perturbants of the lipid phase of the membrane. Thus, SNARE function as regulated by the 0-layer is intimately coupled to the lipids, which must rearrange for fusion. PMID- 17400549 TI - Selective amino acid substitutions convert the creatine transporter to a gamma aminobutyric acid transporter. AB - The creatine transporter (CRT) is a member of a large family of sodium-dependent neurotransmitter and amino acid transporters. The CRT is closely related to the gamma-aminobutyric acid (GABA) transporter, GAT-1, yet GABA is not an effective substrate for the CRT. The high resolution structure of a prokaryotic homologue, LeuT has revealed precise details of the substrate binding site for leucine (Yamashita, A., Singh, S. K., Kawate, T., Jin, Y., and Gouaux, E. (2005) Nature 437, 215-223). We have now designed mutations based on sequence comparisons of the CRT with GABA transporters and the LeuT structural template in an attempt to alter the substrate specificity of the CRT. Combinations of two or three amino acid substitutions at four selected positions resulted in the loss of creatine transport activity and gain of a specific GABA transport function. GABA transport by the "gain of function" mutants was sensitive to nipecotic acid, a competitive inhibitor of GABA transporters. Our results show LeuT to be a good structural model to identify amino acid residues involved in the substrate and inhibitor selectivity of eukaryotic sodium-dependent neurotransmitter and amino acid transporters. However, modification of the binding site alone appears to be insufficient for efficient substrate translocation. Additional residues must mediate the conformational changes required for the diffusion of substrate from the binding site to the cytoplasm. PMID- 17400550 TI - Hypoxia-inducible factor-1alpha, a key factor in the keratinocyte response to UVB exposure. AB - Hypoxia-inducible factor-1 (HIF-1) is a major transcription factor sensitive to oxygen levels, which responds to stress factors under both hypoxic and nonhypoxic conditions. UV irradiation being a common stressor of skin, we looked at the effect of UVB on HIF-1alpha expression in keratinocytes. We found that UVB induces a biphasic HIF-1alpha variation through reactive oxygen species (ROS) generation. Whereas rapid production of cytoplasmic ROS down-regulates HIF-1alpha expression, delayed mitochondrial ROS generation results in its up-regulation. Indeed, activation of p38 MAPK and JNK1 mediated by mitochondrial ROS were required for HIF-1alpha phosphorylation and accumulation after UVB irradiation. Our experiments also revealed a key role of HIF-1alpha in mediating UVB-induced apoptosis. We conclude that the broad impact of the HIF-1 transcription factor on gene expression could make it a key regulator of UV-responsive genes and photocarcinogenesis. PMID- 17400551 TI - Identification of the substrate binding sites within the yeast mitochondrial citrate transport protein. AB - The objective of the present investigation was to identify the substrate binding site(s) within the yeast mitochondrial citrate transport protein (CTP). Our strategy involved kinetically characterizing 30 single-Cys CTP mutants that we had previously constructed based on their hypothesized importance in the structure-based mechanism of this carrier. As part of these studies, a modified transport assay was developed that permitted, for the first time, the accurate determination of K(m) values that were elevated >100-fold compared with the Cys less control value. We identified 10 single-Cys CTP mutants that displayed sharply elevated K(m) values (i.e. 5 to >300-fold). Each of these mutants displayed V(max) values that were reduced by > or = 98% and resultant catalytic efficiencies that were reduced by > or = 99.9%. Importantly, superposition of this functional data onto the three-dimensional homology-modeled CTP structure, which we previously had developed, revealed that nine of these ten residues form two topographically distinct clusters. Additional modeling showed that: (i) each cluster is capable of forming numerous hydrogen bonds with citrate and (ii) the two clusters are sufficiently distant from one another such that citrate is unlikely to interact with all of these residues at the same time. We deduced from these findings that the CTP contains at least two citrate binding sites per monomer, which are located at increasing depths within the translocation pathway. The identification of these sites, combined with an initial assessment of the citrate-amino acid side-chain interactions that may occur at these sites, substantially extends our understanding of CTP functioning at the molecular level. PMID- 17400552 TI - Recognition of hyaluronan released in sterile injury involves a unique receptor complex dependent on Toll-like receptor 4, CD44, and MD-2. AB - Inflammation under sterile conditions is not well understood despite its importance in trauma and autoimmune disease. To investigate this process we established mouse models of sterile injury and explored the role of hyaluronan in mediating inflammation following injury. The response of cultured monocytes to hyaluronan was different than the response to lipopolysaccharide (LPS) despite both being dependent on Toll-like receptor 4 (TLR4). Cultured cells exposed to hyaluronan showed a pattern of gene induction that mimics the response seen in mouse skin after sterile injury with an increase in molecules such as transforming growth factor-beta2 and matrix metalloproteinase-13. These factors were not induced by LPS despite the mutual dependence of both hyaluronan and LPS on TLR4. Explanation for the unique response to hyaluronan was provided by observations that a lack of TLR4 or CD44 in mice diminished the response to sterile injury, and together with MD-2, was required for responsiveness to hyaluronan in vitro. Thus, a unique complex of TLR4, MD-2, and CD44 recognizes hyaluronan. Immunoprecipitation experiments confirmed the physical association of TLR4 and CD44. Taken together, our results define a previously unknown mechanism for initiation of sterile inflammation that involves recognition of released hyaluronan fragments as an endogenous signal of tissue injury. PMID- 17400553 TI - The mobile thylakoid phosphoprotein TSP9 interacts with the light-harvesting complex II and the peripheries of both photosystems. AB - The localization of the plant-specific thylakoid-soluble phosphoprotein of 9 kDa, TSP9, within the chloroplast thylakoid membrane of spinach has been established by the combined use of fractionation, immunoblotting, cross-linking, and mass spectrometry. TSP9 was found to be exclusively confined to the thylakoid membranes, where it is enriched in the stacked grana membrane domains. After mild solubilization of the membranes, TSP9 migrated together with the major light harvesting antenna (LHCII) of photosystem II (PSII) and with PSII-LHCII supercomplexes upon separation of the protein complexes by either native gel electrophoresis or sucrose gradient centrifugation. Studies with a cleavable cross-linking agent revealed the interaction of TSP9 with both major and minor LHCII proteins as identified by mass spectrometric sequencing. Cross-linked complexes that in addition to TSP9 contain the peripheral PSII subunits CP29, CP26, and PsbS, which form the interface between LHCII and the PSII core, were found. Our observations also clearly suggest an interaction of TSP9 with photosystem I (PSI) as shown by both immunodetection and mass spectrometry. Sequencing identified the peripheral PSI subunits PsaL, PsaF, and PsaE, originating from cross-linked protein complexes of around 30 kDa that also contained TSP9. The distribution of TSP9 among the cross-linked forms was found to be sensitive to conditions such as light exposure. An association of TSP9 with LHCII as well as the peripheries of the photosystems suggests its involvement in regulation of photosynthetic light harvesting. PMID- 17400554 TI - Regulation of protein phosphatase inhibitor-1 by cyclin-dependent kinase 5. AB - Inhibitor-1, the first identified endogenous inhibitor of protein phosphatase 1 (PP-1), was previously reported to be a substrate for cyclin-dependent kinase 5 (Cdk5) at Ser67. Further investigation has revealed the presence of an additional Cdk5 site identified by mass spectrometry and confirmed by site-directed mutagenesis as Ser6. Basal levels of phospho-Ser6 inhibitor-1, as detected by a phosphorylation state-specific antibody against the site, existed in specific regions of the brain and varied with age. In the striatum, basal in vivo phosphorylation and dephosphorylation of Ser6 were mediated by Cdk5, PP-2A, and PP-1, respectively. Additionally, calcineurin contributed to dephosphorylation under conditions of high Ca2+. In biochemical assays the function of Cdk5 dependent phosphorylation of inhibitor-1 at Ser6 and Ser67 was demonstrated to be an intramolecular impairment of the ability of inhibitor-1 to be dephosphorylated at Thr35; this effect was recapitulated in two systems in vivo. Dephosphorylation of inhibitor-1 at Thr35 is equivalent to inactivation of the protein, as inhibitor-1 only serves as an inhibitor of PP-1 when phosphorylated by cAMP dependent kinase (PKA) at Thr35. Thus, inhibitor-1 serves as a critical junction between kinase- and phosphatase-signaling pathways, linking PP-1 to not only PKA and calcineurin but also Cdk5. PMID- 17400555 TI - Essential requirement for sphingosine kinase 2 in a sphingolipid apoptosis pathway activated by FTY720 analogues. AB - The clinical immunosuppressant FTY720 is a sphingosine analogue that, once phosphorylated by sphingosine kinase 2 (Sphk2), is an agonist of multiple receptor subtypes for sphingosine 1-phosphate. Short exposures to FTY720 afford long term protection in lymphoproliferative and autoimmune disease models, presumably by inducing apoptosis in subsets of cells essential for pathogenesis. Sphingosine itself is pro-apoptotic, and apoptosis induced with FTY720 or sphingosine is thought to proceed independently of their phosphorylation. Following chemical mutagenesis of Jurkat cells we isolated mutants that are selectively resistant to FTY720 analogue AAL(R), as well as natural sphingolipid bases, including sphingosine. Cells lacking functional Sphk2 were resistant to apoptosis induced with AAL(R), indicating that apoptosis proceeds through AAL(R) phosphorylation. Phosphorylation of AAL(R) was also required for induction of lymphocyte apoptosis in mice, as apoptosis was not induced with the non phosphorylatable chiral analogue, AAL(S). Apoptosis was induced in the spleen but not the thymus of mice administered 1 mg/kg AAL(R), correlating with levels of AAL(R)-phosphate (AFD(R)) in organ extracts. AFD(R) did not induce apoptosis when added to the cell culture medium, indicating that it induces apoptosis through an intracellular target. NBD-labeled AAL(R) localized to the endoplasmic reticulum, and AAL(R) treatment resulted in elevated cytosolic calcium, Bax redistribution from cytosol to mitochondrial and endoplasmic reticulum membranes, and caspase independent mitochondrial permeabilization in Jurkat cells. We therefore describe an apoptotic pathway triggered by intracellular accumulation of sphingolipid base phosphates and suggest that sphingoid base substrates for Sphk2 acting intracellularly could be useful in the treatment of lymphoproliferative diseases. PMID- 17400556 TI - REKLES is an ARID3-restricted multifunctional domain. AB - Bright/Dril1/ARID3a is a B cell-specific, matrix association (or attachment) region-binding transcriptional regulator of immunoglobulin heavy chain genes and of E2F1-dependent cell cycle progression. Bright contains a central DNA binding domain termed ARID (AT-rich interacting domain) and a C-terminal region termed REKLES (for a conserved amino acid motif). The ARID domain has been identified in seven highly conserved families of metazoan proteins (ARID1-5 and JARID1-2), whereas REKLES is found only in the ARID3 subfamily (composed of Bright/ARID3a, Bdp/ARID3b, and Bright-like/ARID3c). REKLES consists of two subdomains: a modestly conserved N-terminal REKLESalpha and a highly conserved (among ARID3 orthologous proteins) C-terminal REKLESbeta. Previously we showed that Bright undergoes nucleocytoplasmic shuttling and that REKLESalpha and -beta were required, respectively, for nuclear import and Crm1-dependent nuclear export. Here we show that Bright further requires REKLESbeta for self-association or paralogue association and for nuclear matrix targeting. REK-LES promotes and regulates the extent of Bright multimerization, which occurs in the absence or presence of target DNA and is necessary for specific DNA binding. REKLESbeta mediated interaction of Bright with Bdp, which localizes strictly to the nucleus, traps Bright within the nucleus via neutralization of its nuclear export activity. These results identify REKLES as a multifunctional domain that has co evolved with and regulates functional properties of the ARID3 DNA binding domain. PMID- 17400557 TI - Structure-function relationships of the viral RNA-dependent RNA polymerase: fidelity, replication speed, and initiation mechanism determined by a residue in the ribose-binding pocket. AB - Studies of the RNA-dependent RNA polymerase (RdRp) from poliovirus (PV), 3Dpol, have shown that Asn-297 permits this enzyme to distinguish ribose from 2' deoxyribose. All animal RNA viruses have Asn at the structurally homologous position of their polymerases, suggesting a conserved function for this residue. However, all prokaryotic RNA viruses have Glu at this position. In the presence of Mg2+, the apparent affinity of Glu-297 3Dpol for 2'-deoxyribonucleotides was decreased by 6-fold relative to wild type without a substantial difference in the fidelity of 2'-dNMP incorporation. The fidelity of ribonucleotide misincorporation for Glu-297 3Dpol was reduced by 14-fold relative to wild type. A 4- to 11-fold reduction in the rate of ribonucleotide incorporation was observed. Glu-297 PV was unable to grow in HeLa cells due to a replication defect equivalent to that observed for a mutant PV encoding an inactive polymerase. Evaluation of the protein-(VPg)-primed initiation reaction showed that only half of the Glu-297 3Dpol initiation complexes were capable of producing VPg-pUpU product and that the overall yield of uridylylated VPg products was reduced by 20 fold relative to wild-type enzyme, a circumstance attributable to a reduced affinity for UTP. These studies identify the first RdRp derivative with a mutator phenotype and provide a mechanistic basis for the elevated mutation frequency of RNA phage relative to animal RNA viruses observed in culture. Although protein primed initiation and RNA-primed elongation complexes employ the same polymerase active site, the functional differences reported here imply significant structural differences between these complexes. PMID- 17400558 TI - Alport syndrome and the X chromosome: implications of a diagnosis of Alport syndrome in females. PMID- 17400559 TI - Diabetes mellitus after kidney transplantation: a French multicentre observational study. AB - BACKGROUND: New-onset diabetes mellitus (NODM)-a common complication of kidney transplantation-is associated with increases in graft loss, morbidity and mortality. METHODS: This is a purely observational study of 527 patients taking a calcineurin inhibitor (CNI), based on data collected at a single routine visit 6 24 months after kidney transplantation. Diabetes was defined according to ADA/WHO guidelines. RESULTS: The mean age of the patients was 47.2 years and 61.1% were men; 49.5% were receiving cyclosporine microemulsion (CsA-ME) and 50.5% tacrolimus (Tac). NODM developed in 7.0% after a median interval of 1.6 months. In CsA-ME-treated patients, the unadjusted cumulative risks of NODM were 5.5% and 8.4% at 1- and 2-year post-transplantation, while in Tac-treated patients, the risks were respectively 17.4% and 21%. Four independent risk factors (RFs) were identified by multivariate analysis: maximum lifetime body mass index>25 [odds ratio (OR)=5.1], pre-transplantation impaired fasting glucose (OR=4.7), hepatitis C status (OR=4.7) and Tac vs CsA-ME treatment (OR=3.0). CONCLUSIONS: NODM is associated with certain RFs present prior to kidney transplantation, and with treatment with Tac as opposed to CsA-ME. PMID- 17400560 TI - Identification of current smokers among renal transplant recipients. AB - BACKGROUND: In renal transplant recipients, smoking is associated with a high burden of cardiovascular disease and a higher risk of graft loss. Surprisingly, the results of measurement of cotinine serum level, the gold standard for the detection of active smoking, have not been confronted with self-reported smoking history in this group. The aim of our study was to identify and characterize the smoking group of renal transplant recipients. METHODS: Cotinine serum level was measured and all patients were asked to fill out an anonymous questionnaire on smoking history. RESULTS: Out of 233 renal transplant recipients, 106 (45%) reported to be lifetime and current non-smokers: cotinine serum level was below detection limit in all; among the 127 renal transplant recipients (55%) with a lifetime history of smoking, cotinine level was diagnostic of current smoking in 32 (25%). Only 21 of the current smokers (66%) declared to the nephrologist that they had continued smoking whereas 11 (34%) claimed to be non-smokers. Current smokers were younger (P=0.01) than former smokers. CONCLUSION: The identification of current smokers among renal transplant recipients should start with questioning about lifetime history of smoking and if positive, measurement of cotinine serum level. Indeed up to 34% of current smokers do not acknowledge they are active smokers and would otherwise not offer to participate in programmes to stop smoking. PMID- 17400561 TI - Differences in heart rate variability during haemodialysis and haemofiltration. AB - BACKGROUND: The aim of our study was to evaluate whether convective (haemofiltration, Hf) and diffusive (haemodialysis, Hd) dialysis techniques induce different patterns of long- and short-term autonomic adjustments in haemodynamically stable dialysis patients. METHODS: Ten haemodynamically stable Hd patients were studied. Each patient underwent a block of six Hd sessions, then was switched to six Hf. During the last session of each dialytic treatment, continuous beat to beat measurements of systolic arterial pressure (SAP) and heart rate (HR) were performed. Spectral analysis of heart rate variability (HRV) was made before and during the treatment to evaluate the modification of autonomic nervous system activity. RESULTS: Baseline values of plasma sodium, body weight, HR and SAP were not different for the two considered methods of dialysis, while the baseline values of normalized LF were significantly higher in Hf as compared to Hd and the opposite was observed for HF powers (P < 0.001). Sodium balance and body weight loss per hour did not differ between Hd and Hf while body temperature was kept constant in all sessions. Throughout the dialytic procedures, with both techniques, SAP was constant, while HR diminished from the first hour till the end of the procedure (P < 0.05). An increase in LF (and decrease in HF) was noticed only in the case of Hd, considering normalized units (P < 0.05). These selective changes were maintained also during the recovery after the procedure. CONCLUSIONS: The spectral analysis of RR interval variability during Hd and Hf suggests a potential autonomic advantage with Hf, to be added to the well-recognized intrinsic greater haemodynamic stability. PMID- 17400562 TI - Delaying initiation of dialysis till symptomatic uraemia--is it too late? AB - BACKGROUND: The optimal timing of initiating renal replacement therapy in patients with chronic renal failure remains uncertain. The primary objective of our study is to determine whether delaying dialysis initiation as a result of patients' choice may have any impact on survival in subjects with end-stage renal disease. METHODS: We prospectively studied the clinical outcome during the first year of all consecutive patients (n=233) deemed suitable for peritoneal dialysis (PD) after pre-dialysis counselling over a 2-year period from 2002 to 2004. All patients who were offered dialysis were included in the analysis from the day of initial counselling regardless of whether or not they were eventually established on PD. RESULTS: There were 151 'elective starters' (50.3% male, mean+/-SD age=57.7+/-13.9 years, 39.7% diabetic) who were electively initiated on dialysis when glomerular filtration rate reached 10 ml/min/1.73 m2 or below. The remaining 82 subjects (53.7% male, mean+/-SD age=58.4+/-11.3 years, 46.3% diabetic, P=0.33 vs elective starters) declined dialysis initially (initial refusers). On follow up, 45 (55%) initial refusers developed a uraemic emergency and agreed to undergo dialysis, and 39 (48%) were eventually established on maintenance PD (late starters). Kaplan-Meier analysis of 1-year survival showed a significantly higher rate of all-cause (18.3% vs 6.6%, P=0.004, log-rank test) and cardiovascular (9.8% vs 2.6%, P=0.014) mortality among the initial refusers. CONCLUSION: Patients who refuse timely start of dialysis have worse overall outcome at one year after the offer of dialysis, compared with elective starters. PMID- 17400563 TI - Efficacy of renal artery angioplasty and stenting in a solitary functioning kidney. AB - OBJECTIVE: The aim of this study was to share our experience of percutaneous renal artery angioplasty and stenting (PTRAS) in our patients with a solitary functioning kidney over a 10-year period. METHODS: The procedures were performed on 75 patients from 1995 to 2004. Data were collected retrospectively from case notes of patients. The definition for solitary functioning kidney was a contralateral kidney size of <8 cm, complete occlusion of contralateral renal artery or previous nephrectomy. Serum creatinine was considered improved or worse if the deviation from the baseline value was >20%. RESULTS: For the purpose of halting renal deterioration (n=47), there were improvement and stabilization in 21% and 55% at 3 months and 28% and 28% at 12 months. Systolic blood pressure (n=27) improvement and stabilization were achieved in 33% and 56% both at 3 and 12 months. Diastolic blood pressure (n=27) improvement and stabilization were 22% and 70% at 3 months and 33% and 48% at 12 months. Five out of seven patients with acute renal failure (serum creatinine>500 micromol/l and requiring haemodialysis) pre-procedure were dialysis-free at 12 months. Complications occurred in 19 (25%) patients and these included bleeding (n=16), pseudoaneurysm (n=3), renal artery dissection (n=2) and cholesterol embolization (n=1). CONCLUSION: PTRAS in a solitary functioning kidney produced clinical benefits in the majority of patients with resistant hypertension and renal function deterioration. PMID- 17400564 TI - Use of ultrafiltered dialysate is associated with improvements in haemodialysis associated morbidity in patients treated with reused dialysers. AB - BACKGROUND: Morbidity in haemodialysis patients is associated with chronic inflammation. Microbiological contaminants derived from dialysate are thought to be one inflammatory stimulus and previous studies found that highly purified dialysate reduces inflammation and morbidity. These studies were performed in the absence of practices, such as dialyser reuse, that are potentially inflammatory. We tested the hypothesis that highly purified dialysate reduces inflammation and morbidity even in the presence of other inflammatory stimuli. METHODS: This was a prospective observational study. After obtaining baseline data on inflammation, oxidant stress, nutrition and anaemia correction with standard dialysate, 105 patients were switched to dialysate that was ultrafiltered at the point of use and follow-up data were collected at 3-month intervals for 12 months. RESULTS: Introduction of ultrafiltered dialysate did not significantly reduce inflammation, as assessed by plasma concentrations of C-reactive protein and interleukin-6 or oxidant stress, as assessed by plasma concentrations of protein carbonyls and protein-free sulphydryls. Neither did it improve anaemia correction, as assessed by plasma haemoglobin and erythropoietin dose. However, introduction of ultrafiltered dialysate was associated with a significant reduction in plasma beta(2)-microglobulin concentration and a significant improvement in nutritional status, assessed by plasma albumin concentration and creatinine generation rate as a marker of muscle mass. CONCLUSION: Use of ultrafiltered dialysate was associated with improvements in some measures of morbidity, such as plasma beta(2)-microglubulin and nutrition. These changes occurred in spite of the presence of inflammatory stimuli, such as dialyser reuse, and with no measurable reduction in inflammation and oxidant stress. PMID- 17400565 TI - N-acetylcysteine attenuates NSAID-induced rat renal failure by restoring intrarenal prostaglandin synthesis. AB - BACKGROUND: Renal failure is a threatening side-effect of NSAID administration, consequent to NSAID-mediated abrogation of prostaglandin synthesis and resultant renal ischaemia. N-acetylcysteine (NAC) has renoprotective properties. We examined effects of NAC in a rat model of NSAID-induced renal failure. METHODS: Renal failure was generated in 80 rats by 6-day water deprivation and 3-day 15 mg/kg/day diclofenac injection. The rats were concomitantly treated, or not, by NAC, 40 mg/kg/day. Renal function was evaluated by cystatin C, creatinine and urea. Intrarenal blood flow was measured by laser Doppler. The kidneys were subjected to pathological examination or evaluation of intrarenal NO, H2O2 and PGE2. RESULTS: NAC significantly attenuated deterioration of renal function in diclofenac-treated rats: cystatin C dropped from 2.8+/-0.35 to 2.2+/-0.67 mg/l, P=0.016; creatinine from 1.2+/-0.97 to 0.96+/-0.19 mg/dl, P=0.02; urea from 208.4+/-57.9 to 157.6+/-33.7 mg/dl, P=0.028. Diclofenac-inflicted hystopathological damage was significantly reduced following NAC treatment. Intrarenal medullar blood flow dropped by 51+/-12.4% in diclofenac-treated rats, but only by 14+/-3.39% in those receiving NAC after diclofenac injection (P<0.001). H2O2 was elevated in renal tissues of diclofenac-receiving rats, while decreased in NAC-treated animals. PGE2 release by diclofenac-treated rats dropped significantly, but was restored after NAC administration both in renal cortices (144.7+/-10.4 vs 19.7+/-1.5 pmol/ml, P<0.001) and medullae (148.5+/-7.3 vs 66.6+/ 7.3 pmol/ml, P<0.001). CONCLUSIONS: In this model of renal failure induced by NSAID administration combined with water deprivation, NAC treatment successfully attenuated the deterioration of renal function by inducing renal vasodilatation, decreasing oxidative stress via inhibition of intrarenal ROS content and restoration of intrarenal PGE2 release back to the basal levels. PMID- 17400566 TI - Unexpected and late diagnosis (28th week) of pregnancy in a 39-year-old patient on chronic haemodialysis. PMID- 17400567 TI - Implementation of a vascular access quality programme improves vascular access care. AB - INTRODUCTION: In the Netherlands an access quality improvement plan (QIP) was introduced by vascular access coordinators (VAC) with the aim to decrease vascular access-related complications by preemptive intervention of malfunctioning accesses. A vascular access QIP was established in 24 centres (46% of all Dutch facilities) and a structural multidisciplinary vascular access meeting was instituted. In these centres, including 2300 patients, a protocol for enhancement of fistula creation and access surveillance programme was implemented, with instruction of physicians and nurses, and rounds to discuss complications and evaluate vascular access interventions. The number and type of vascular access, permanent catheters, thrombosis rates and number of interventions were evaluated at the start and end of the study period. RESULTS: After the surveillance programme, the number of autogenous arterio-venous fistulas (AVFs) had increased significantly from 69 to 77% (P < 0.01), while the use of temporary subclavian vein catheters declined (34% vs 11%) (P < 0.01), with a substantially higher percentage of jugular vein catheters (from 23 to 35%). Interventional treatment of malfunctioning accesses by percutaneous transluminal angioplasty (PTA) (from 0.39 to 0.50 patient/year; P < 0.001)) and surgical revisions (from 0.06 to 0.12 per patient/year; P < 0.001) also increased. CONCLUSION: These data demonstrate that a vascular access QIP resulted in placement of more autogenous AVFs, increased number of PTAs and surgical interventions. These findings suggest that a vascular access care QIP is worthwhile to improve dialysis patients' care and access morbidity. PMID- 17400568 TI - When can we really diagnose idiopathic retroperitoneal fibrosis? PMID- 17400569 TI - Estimating population size with two- and three-stage sampling designs. AB - Reliable estimates of population size are important for developing and monitoring health programs in at-risk populations. Laska, Meisner, and Siegel (Biometrics 1988;44:461-72) developed an unbiased estimator for the size of a population at a single venue based on a single sample. Because many populations of interest are not contained within a single venue, this article generalizes the Laska, Meisner, and Siegel estimator to incorporate two- and three-stage sampling designs and enable estimation of total population size over multiple venues. Use of the estimator with two- and three-stage sampling designs is illustrated with examples that estimate the size of a population of individuals who socialize over a 4-week period at public venues where transmission of human immunodeficiency virus and other sexually transmitted infections is likely to occur. PMID- 17400570 TI - Racial discrimination and breast cancer incidence in US Black women: the Black Women's Health Study. AB - Perceived discrimination may contribute to somatic disease. The association between perceived discrimination and breast cancer incidence was assessed in the Black Women's Health Study. In 1997, participants completed questions on perceived discrimination in two domains: "everyday" discrimination (e.g., being treated as dishonest) and major experiences of unfair treatment due to race (job, housing, and police). Cox proportional hazards models were used to estimate incidence rate ratios, controlling for breast cancer risk factors. From 1997 to 2003, 593 incident cases of breast cancer were ascertained. In the total sample, there were weak positive associations between cancer incidence and everyday and major discrimination. These associations were stronger among the younger women. Among women aged less than 50 years, those who reported frequent everyday discrimination were at higher risk than were women who reported infrequent experiences. In addition, the incidence rate ratio was 1.32 (95% confidence interval: 1.03, 1.70) for those who reported discrimination on the job and 1.48 (95% confidence interval: 1.01, 2.16) for those who reported discrimination in all three situations - housing, job, and police - relative to those who reported none. These findings suggest that perceived experiences of racism are associated with increased incidence of breast cancer among US Black women, particularly younger women. PMID- 17400571 TI - Nurse staffing level and nosocomial infections: empirical evaluation of the case crossover and case-time-control designs. AB - The authors compared a case-crossover design, a case-time-control design, and a cohort design to evaluate the effect of nurse staffing level on the risk of nosocomial infections. They evaluated two strategies, conditional logistic regression and generalized estimating equation, to analyze the case-crossover study. The study was performed among critically ill patients in the medical intensive care unit of the University of Geneva Hospitals, Geneva, Switzerland. Of 366 patients who stayed more than 7 days in the intensive care unit between 1999 and 2002, 144 developed an infection. The main reasons for admission were infectious (35.3%), cardiovascular (32.5%), and pulmonary (19.7%) conditions. A comparison of the three study designs showed that lower nurse staffing was associated with an approximately 50% increased risk of nosocomial infections. All analyses yielded similar estimates, except that the point estimate obtained by the conditional logistic regression used in the case-crossover design was biased away from unity; the generalized estimating equation yielded unbiased results and is the most appropriate technique for case-crossover designs. The case-crossover methodology in hospital epidemiology is a promising alternative to traditional approaches, but selection of the referent periods is challenging. PMID- 17400572 TI - An empirical codon model for protein sequence evolution. AB - In the past, 2 kinds of Markov models have been considered to describe protein sequence evolution. Codon-level models have been mechanistic with a small number of parameters designed to take into account features, such as transition transversion bias, codon frequency bias, and synonymous-nonsynonymous amino acid substitution bias. Amino acid models have been empirical, attempting to summarize the replacement patterns observed in large quantities of data and not explicitly considering the distinct factors that shape protein evolution. We have estimated the first empirical codon model (ECM). Previous codon models assume that protein evolution proceeds only by successive single nucleotide substitutions, but our results indicate that model accuracy is significantly improved by incorporating instantaneous doublet and triplet changes. We also find that the affiliations between codons, the amino acid each encodes and the physicochemical properties of the amino acids are main factors driving the process of codon evolution. Neither multiple nucleotide changes nor the strong influence of the genetic code nor amino acids' physicochemical properties form a part of standard mechanistic models and their views of how codon evolution proceeds. We have implemented the ECM for likelihood-based phylogenetic analysis, and an assessment of its ability to describe protein evolution shows that it consistently outperforms comparable mechanistic codon models. We point out the biological interpretation of our ECM and possible consequences for studies of selection. PMID- 17400573 TI - Evolution of population structure in a highly social top predator, the killer whale. AB - Intraspecific resource partitioning and social affiliations both have the potential to structure populations, though it is rarely possible to directly assess the impact of these mechanisms on genetic diversity and population divergence. Here, we address this for killer whales (Orcinus orca), which specialize on prey species and hunting strategy and have long-term social affiliations involving both males and females. We used genetic markers to assess the structure and demographic history of regional populations and test the hypothesis that known foraging specializations and matrifocal sociality contributed significantly to the evolution of population structure. We find genetic structure in sympatry between populations of foraging specialists (ecotypes) and evidence for isolation by distance within an ecotype. Fitting of an isolation with migration model suggested ongoing, low-level migration between regional populations (within and between ecotypes) and small effective sizes for extant local populations. The founding of local populations by matrifocal social groups was indicated by the pattern of fixed mtDNA haplotypes in regional populations. Simulations indicate that this occurred within the last 20,000 years (after the last glacial maximum). Our data indicate a key role for social and foraging behavior in the evolution of genetic structure among conspecific populations of the killer whale. PMID- 17400574 TI - The Yersinia kristensenii O11 O-antigen gene cluster was acquired by lateral gene transfer and incorporated at a novel chromosomal locus. AB - We have sequenced the O-antigen gene clusters for the Escherichia coli O98 and Yersinia kristensenii O11 O antigens. The basic structures of these O antigens are identical, and the sequence data indicate that Y. kristensenii O11 gained its O-antigen gene cluster by lateral gene transfer (LGT). Escherichia coli O98 has a typical O-antigen gene cluster between galF and gnd as is usual in E. coli. However, the O-antigen gene cluster of Y. kristensenii O11 is not located at the traditional Yersinia O-antigen gene cluster locus, between hemH and gsk, but at a novel chromosomal locus between aroA and cmk where it is flanked by remnant galF and gnd genes that indicate the probable source of the gene cluster. Phylogenetic analysis indicated that the source was not E. coli itself but a species in the Escherichia, Salmonella, and Klebsiella group of genera. Although other O-antigen studies imply LGT on the basis of the hypervariability of the loci and GC content, this report also identifies a potential donor and provides evidence for the mechanism involved. Remnant insertion sequence (IS) sequences flank the galF and gnd remnants and suggest that LGT of the gene cluster was IS mediated. PMID- 17400575 TI - Non-professional health practitioners and referrals to facilities: lessons from maternal care in Bangladesh. AB - Over half a million women in the developing world die of pregnancy and childbirth related causes each year, despite well-known interventions to manage most maternal complications. One problem facing policy makers is that women in low income settings often seek care from a range of non-professional sources when they have trouble with pregnancy and childbirth. Questions remain as to the best way to engage with such providers to encourage use of professional care, in part because little policy-oriented research has attempted to study the roles of non professional practitioners, and the specific situations which can encourage or discourage referral behaviour. This paper investigates the roles played by alternative health practitioners in referral to facilities for maternal care in Bangladesh. In-depth case studies were used to investigate labour experiences, decision-making processes and the roles played by key individuals in deciding to use professional services. Findings show that the commonly used heading of 'traditional birth attendant' is often too broad for programmatic use, as it encompasses a range of individuals with different reasons to work with, or oppose, professional services. It was found that women seek care from multiple non-professional cadres who each have differing services, scopes and linkages to professional care. Policy makers need to understand the roles of different providers and potential links to professional care which can be built upon to encourage the use of professional emergency care for maternal complications in low-income settings. PMID- 17400576 TI - A rapid assessment methodology for the evaluation of primary care organization and performance in Brazil. AB - INTRODUCTION: This study presents a methodology for the rapid assessment of the organization and performance of primary care services at district level. It compares results from an earlier provider survey in Petropolis, Brazil with those obtained directly from users of two types of primary care services (newly developed Family Health Program services or 'PSF'--Programa de Saude da Familia- and traditional services) in the same municipality. The aim is to demonstrate the validity of the methodology and its potential use in measuring and improving district-level primary care services in developing countries. METHODS: The study adapted a previously developed questionnaire to measure essential dimensions of primary care. Users (n = 468) were randomly selected from each of the 40 primary care clinics in the district and administered a question survey. Responses were used to create nine measures: an overall composite 'total primary care index' and eight sub-indices each pertaining to an essential primary care dimension. RESULTS: Primary care services show considerable variation in user experiences. Users of the new (PSF) clinics reported higher overall assessments of the total primary care index and the sub-indices for gatekeeping, comprehensiveness, family focus and community orientation than did users of traditional services (P < 0.05). The total primary care score was internally consistent with a Cronbach's alpha of 0.8, and could be reduced to only one principal component. User assessments of primary care services were predicted by self-rated health (OR 1.72) and the site of care (OR 1.03). User and provider assessments of the total primary care index were not significantly different, disagreeing on only two sub indices (gatekeeping and family focus, P < 0.05). CONCLUSIONS: The study presents a rapid and valid method of obtaining information about clinic-level variation in primary care organization and performance at district level. The total primary care index was not sensitive to demographic or socio-economic characteristics of clients, was internally consistent and appears valid given similar results obtained using two different data sources. With some adaptation the instrument could potentially be applied in other developing countries. PMID- 17400577 TI - 'We are also dying like any other people, we are also people': perceptions of the impact of HIV/AIDS on health workers in two districts in Zambia. AB - In countries with a high AIDS prevalence, the health workforce is affected by AIDS in several ways. In Zambia, which has a prevalence rate of 16.5%, a study was carried out in 2004 with the aim to: explore the impact of HIV/AIDS on health workers, describe their coping mechanisms and recommend supportive measures. The qualitative study was complemented by a survey using self-administered questionnaires in four selected health facilities in two rural districts in Zambia, Mpika and Mazabuka. It is one of the few studies to have explored the impact of HIV/AIDS from the perspective of health workers and managers in the region. Thirty-four in-depth interviews and five group discussions were conducted with health workers, managers and volunteers, and 82 self-administered questionnaires were filled out by health workers. In addition, burnout among 42 health workers was measured using the Maslach Burnout Inventory (MBI). The MBI measures three components that contribute to burnout: emotional exhaustion, depersonalization and personal accomplishment. The results show that in both districts, HIV/AIDS has had a negative impact on workload and has considerably changed or added tasks to already overburdened health workers. In Mpika, 76% of respondents (29/38), and in Mazabuka, 79% (34/44) of respondents, expressed fear of infection at the workplace. HIV-positive health workers remained 'in hiding', did not talk about their illness and suffered in silence. Despite the fact that health workers were still relatively motivated, emotional exhaustion occurred among 62% of the respondents (26/42). The interviews revealed that counsellors and nurses were especially at risk for emotional exhaustion. In each of the selected facilities, organizational support for health workers to deal with HIV/AIDS was either haphazardly in place or not in place at all. AIDS complicates the already difficult work environment. In addition to health workers, management also needs support in dealing with AIDS at the workplace. PMID- 17400578 TI - Elevation of 8-hydroxydeoxyguanosine in DNA from isolated mouse lung cells following in vivo treatment with aflatoxin B(1). AB - Aflatoxin B(1) (AFB(1)) is a mycotoxin produced by some strains of Aspergillus and is a recognized pulmonary and hepatic carcinogen. The most widely accepted mechanism of AFB(1) carcinogenicity involves bioactivation to AFB(1)-8,9-exo epoxide and binding to DNA to form AFB(1)-N(7)-guanine. Another potential cause of DNA damage is AFB(1)-mediated stimulation of reactive oxygen species formation, leading to oxidation of DNA bases. The objective of this study was to determine the ability of AFB(1) to cause oxidative DNA damage in lung cell types of the A/J mouse. The formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG) in freshly isolated mouse lung alveolar macrophages, alveolar type II cells, and nonciliated bronchial epithelial (Clara) cells was assessed by high-performance liquid chromatography with electrochemical detection. An approximately 3-fold increase in 8-OHdG formation occurred in both alveolar macrophage and Clara cell preparations isolated from A/J mice 2 h following treatment with a single tumorigenic dose of 50 mg/kg AFB(1) ip (n = 3, p < 0.05). Prior treatment with 300 kU/kg polyethylene glycol-conjugated catalase prevented the AFB(1)-induced increase in 8-OHdG levels in all mouse lung cell preparations (n = 3, p < 0.05). These results support the possibility that oxidative DNA damage in mouse lung cells contributes to AFB(1) carcinogenicity. PMID- 17400579 TI - Effects of low concentrations of arsenic on the innate immune system of the zebrafish (Danio rerio). AB - Arsenic has been associated with a multitude of human health problems; however, its impact on host resistance to infection has not been extensively researched. In vertebrates, the innate immune response is vital for potentiating the adaptive immune response. Therefore, dampening of the innate immune response results in an immunocompromised host. In this present study, effects of low concentrations of arsenic on zebrafish resistance to infection are evaluated. Exposure to 2 and 10 ppb arsenic, both considered safe levels in drinking water, resulted in a greater than 50-fold increase in viral load and at least a 17-fold increase in bacterial load in embryos. To determine the cause of this amplified pathogen load, important components of the innate immune system were analyzed. Presence of arsenic dampened the overall innate immune health of the fish as evidenced by reductions in respiratory burst activity. Viral infection, after arsenic exposure, showed decreases of up to 13- and 1.5-fold changes in interferon and Mx mRNA expression, respectively. Bacterial infection, post arsenic exposure, demonstrated at least 2.5- and 4-fold declines in interleukin-1beta and tumor necrosis factor-alpha mRNA levels, respectively. Maximum expression of these essential cytokines was also delayed upon arsenic exposure. Our data indicate that arsenic exposure, at concentrations deemed safe in drinking water, suppresses the overall innate immune function in zebrafish and present the zebrafish as a unique model for studying immunotoxicity of environmental toxicants. To our knowledge, this is the first report describing the effects of such low levels of arsenic on host resistance to infection. PMID- 17400580 TI - An L-tyrosine derivative and PPARgamma agonist, GW7845, activates a multifaceted caspase cascade in bone marrow B cells. AB - Apoptosis is a critical event in the deletion of B lymphocytes prior to their migration to the periphery. Synthetic peroxisome proliferator activated receptor gamma (PPARgamma) agonists, including the drug GW7845 and the environmental contaminant mono-(2-ethylhexyl) phthalate, as well as an endogenous ligand, 15 deoxy-Delta(12,14)-prostaglandin J(2), induce clonally unrestricted apoptosis in pro/pre-B cells. Considering that PPARgamma agonists are used clinically for the treatment of diabetes and postulated to be useful as chemotherapeutics, we used GW7845 as a model PPARgamma agonist to examine the mechanism of cell death that may contribute to tumor killing as well as normal bone marrow B lymphocyte toxicity. GW7845 induced rapid mitochondrial membrane depolarization and release of cytochrome c, along with nearly concurrent activation of capases-2, -3, -8, and -9 in primary pro-B cells and BU-11 cells, a nontransformed pro/pre-B cell line. GW7845-induced apoptosis was reduced significantly in Bax-deficient and Apaf-1 mutant primary pro-B cells, supporting the conclusion that GW7845-induced apoptosis is mitochondria- and apoptosome-dependent. Using benzyloxycarbonyl-VAD fluoromethyl ketone (VAD-FMK) as a pan-caspase inhibitor, we demonstrated that an initial cytochrome c release occurred independently of caspase activation and that only caspase-9 activation was partially caspase independent. The attenuation of GW7845-induced apoptosis by multiple FMK-labeled peptide sequences suggests that multiple caspase pathways are responsible for initiating and executing apoptosis. The strong activation of Bid provides a mechanism by which caspases-2, -3, and -8 may amplify the apoptotic signal. These data support the hypothesis that pharmacologic concentrations of PPARgamma agonists induce an intrinsic apoptotic pathway that is driven in normal bone marrow B cells by multiple amplification loops. PMID- 17400581 TI - Alterations in gene expression and testosterone synthesis in the testes of male rats exposed to perfluorododecanoic acid. AB - Perfluorododecanoic acid (PFDoA, C12), a synthetic perfluorinated chemical containing 12 carbons, has broad industrial applications and has been detected in sera from humans and other animals; however, few reports have addressed the effects of PFDoA exposure on male reproduction. In the present study, the effects of PFDoA exposure on testes ultrastructure, testosterone levels, and steroidogenic gene expression were investigated. Male rats were orally dosed for 14 days with 1, 5, or 10 mg PFDoA/kg/day or with vehicle. Absolute testis weight was diminished at the highest dose while relative testes weight was markedly increased at doses of 5 and 10 mg/kg/day. Total serum cholesterol levels were significantly increased at the highest dose. While luteinizing hormone was significantly decreased at the highest dose, testosterone was markedly decreased at doses of 5 and 10 mg PFDoA/kg/day. Serum levels of follicle-stimulating hormone were not significantly affected by PFDoA, and estradiol levels were markedly decreased only at 5 mg/kg/day. Leydig cells, Sertoli cells, and spermatogenic cells from rats that received 5 or 10 mg PFDoA/kg/day, exhibited apoptotic features including dense irregular nuclei, condensed chromatin, ill defined nuclear membranes, and abnormal mitochondria. PFDoA exposure resulted in significant declines in mRNA expression of several genes involved in cholesterol transport and steroid biosynthesis at doses of 5 and 10 mg PFDoA/kg/day, while the gene expression of luteinizing hormone receptor and aromatase was not significantly changed. Our results demonstrate that PFDoA affects the reproduction function of male rats via alterations in steroidogenesis genes, testosterone levels, and testes ultrastructure. PMID- 17400582 TI - Cumulative effects of dibutyl phthalate and diethylhexyl phthalate on male rat reproductive tract development: altered fetal steroid hormones and genes. AB - Exposure to plasticizers di(n-butyl) phthalate (DBP) and diethylhexyl phthalate (DEHP) during sexual differentiation causes male reproductive tract malformations in rats and rabbits. In the fetal male rat, these two phthalate esters decrease testosterone (T) production and insulin-like peptide 3 (insl3) gene expression, a hormone critical for gubernacular ligament development. We hypothesized that coadministered DBP and DEHP would act in a cumulative dose-additive fashion to induce reproductive malformations, inhibit fetal steroid hormone production, and suppress the expression of insl3 and genes responsible for steroid production. Pregnant Sprague Dawley rats were gavaged on gestation days (GD) 14-18 with vehicle control, 500 mg/kg DBP, 500 mg/kg DEHP, or a combination of DBP and DEHP (500 mg/kg each chemical; DBP+DEHP); the dose of each individual phthalate was one-half of the effective dose predicted to cause a 50% incidence of epididymal agenesis. In experiment one, adult male offspring were necropsied, and reproductive malformations and androgen-dependent organ weights were recorded. In experiment two, GD18 testes were incubated for T production and processed for gene expression by quantitative real-time PCR. The DBP+DEHP dose increased the incidence of many reproductive malformations by >or=50%, including epididymal agenesis, and reduced androgen-dependent organ weights in cumulative, dose additive manner. Fetal T and expression of insl3 and cyp11a were cumulatively decreased by the DBP+DEHP dose. These data indicate that individual phthalates with a similar mechanism of action, but with different active metabolites (monobutyl phthalate versus monoethylhexyl phthalate), can elicit dose-additive effects when administered as a mixture. PMID- 17400583 TI - The contribution of methotrexate exposure and host factors on transcriptional variance in human liver. AB - Long-term administration of methotrexate (MTX) for management of chronic inflammatory diseases is associated with risk of liver damage. In this study, we examined the transcriptional profiles of livers from patients treated with MTX. The possibility that expression signatures correlate with grade of fibrosis or underlying rheumatic disease was evaluated. Twenty-seven patients taking MTX were accrued for this study. Ten non-MTX-exposed normal liver specimens were used as controls. Global mRNA expression was assayed using oligonucleotide arrays. A total of 205 genes were significantly altered in MTX-exposed livers. Six of these genes were validated by qPCR. Two genes, CLN8 and ANKH that map to chromosomal locations previously associated with rheumatoid arthritis, were found to be elevated in MTX-exposed samples. Subsequent pathway analysis indicates that MTX exposure is associated with the following key alterations: (1) upregulation of lipid biosynthetic genes, consistent with MTX-induced steatosis, (2) downregulation of proinflammatory chemokines, consistent with the anti inflammatory effects of MTX, and (3) elevation of complement pathway gene expression. Complement 5, shown earlier to be correlated with liver fibrosis in mice, was found to be elevated (twofold) in MTX-exposed livers. In conclusion, we have found the expression of a number of genes associated with rheumatic disease and/or MTX exposure to be significantly different. Differences in complement expression provide the rationale for future correlative studies between MTX induced liver fibrosis and C5 alleles in order to identify patients with increased risk for fibrosis. PMID- 17400584 TI - Quantification of chemical peptide reactivity for screening contact allergens: a classification tree model approach. AB - In the interest of reducing animal use, in vitro alternatives for skin sensitization testing are under development. One unifying characteristic of chemical allergens is the requirement that they react with proteins for the effective induction of skin sensitization. The majority of chemical allergens are electrophilic and react with nucleophilic amino acids. To determine whether and to what extent reactivity correlates with skin sensitization potential, 82 chemicals comprising allergens of different potencies and nonallergenic chemicals were evaluated for their ability to react with reduced glutathione (GSH) or with two synthetic peptides containing either a single cysteine or lysine. Following a 15-min reaction time with GSH, or a 24-h reaction time with the two synthetic peptides, the samples were analyzed by high-performance liquid chromatography. UV detection was used to monitor the depletion of GSH or the peptides. The peptide reactivity data were compared with existing local lymph node assay data using recursive partitioning methodology to build a classification tree that allowed a ranking of reactivity as minimal, low, moderate, and high. Generally, nonallergens and weak allergens demonstrated minimal to low peptide reactivity, whereas moderate to extremely potent allergens displayed moderate to high peptide reactivity. Classifying minimal reactivity as nonsensitizers and low, moderate, and high reactivity as sensitizers, it was determined that a model based on cysteine and lysine gave a prediction accuracy of 89%. The results of these investigations reveal that measurement of peptide reactivity has considerable potential utility as a screening approach for skin sensitization testing, and thereby for reducing reliance on animal-based test methods. PMID- 17400585 TI - Mgat5 and Pten interact to regulate cell growth and polarity. AB - Phosphatase and tensin homolog (Pten) phosphatase opposes intracellular phosphoinositide 3-kinase (PI3K)/Akt signaling and is a potent tumor suppressor, while Golgi beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is positively associated with cancer progression and metastasis. beta1,6GlcNAc-branched N glycans on receptor glycoproteins promote their surface residency and sensitizes cells to growth factor signaling. Here we demonstrate that the Pten heterozygosity in mouse embryonic fibroblasts enhances cell adhesion-dependent PI3K/Akt signaling, cell spreading, and proliferation, while Pten/Mgat5 double mutant cells are normalized. However, planar asymmetry typical of fibroblasts and invasive carcinomas is not fully rescued, suggesting that Mgat5 and Pten function together to regulate the membrane dynamics of PI3K/Akt signaling typical of motile cells. Pten heterozygosity was associated with increased surface beta1,6GlcNAc-branched N-glycans, suggesting positive feedback from PI3K signaling to N-glycan branching. In vivo, Mgat5(-/-) Pten(+/-) and Mgat5(+/ )Pten(+/-)mutant mice showed a small but significant increase in longevity compared with Pten(+/-) mice. Taken together, our results reveal that Mgat5 and Pten interact in an opposing manner to regulate cellular sensitivities to extracelluar growth cues. PMID- 17400586 TI - Spontaneous hybridization between maize and teosinte. AB - The closest wild relatives of maize, Zea mays ssp. mays are various Zea taxa known as "teosinte." Hybrids between maize and the teosinte taxon, Zea mays ssp. mexicana, often occur when the 2 are sympatric in Mexico. Measuring the spontaneous hybridization rate of the 2 taxa would shed light on the mechanisms contributing to the evolution and persistence of these hybrid swarms. We conducted a series of field experiments in Riverside, CA, to measure the natural hybridization rates between maize and 2 teosinte taxa, Z. m. ssp. mexicana and Zea mays ssp. parviglumis. We planted teosinte within and near maize plantations. Hybrids were identified by progeny testing for a maize-specific herbicide resistance allele and a teosinte-specific allozyme allele. Hybridity was confirmed by growing putative hybrid progeny to maturity to evaluate whether they had the characteristic morphology of maize x teosinte hybrids. We found that maize and Z. m. ssp. mexicana naturally hybridize at a low rate (<1%), whereas Z. m. ssp. parviglumis hybridizes with the crop at a high rate (>>50%). PMID- 17400587 TI - Temporal interactions between oral irritants: piperine, zingerone, and capsaicin. AB - Sequential presentation of 2 irritants may produce cross-sensitization or cross adaptation effects upon introduction of the second irritant. In Experiment 1, subjects were given either 34 min of stimulation with zingerone, capsaicin, or piperine or one of those irritants for 23 min followed by blanks for 23 min. In Experiment 2, subjects received one irritant for 23-min irritants, followed immediately by another for 23 min (piperine --> zingerone, piperine --> capsaicin, zingerone --> piperine, or zingerone --> capsaicin). Cross sensitization was observed for the piperine --> zingerone, zingerone --> piperine, and piperine --> capsaicin groups; cross-adaptation was observed for the zingerone --> capsaicin group. Cross-adaptation and cross-sensitization were predicted by adding the independent time courses of the respective irritants, starting the second at the offset of the first. These responses were also predicted by a mathematical model of central processing of primary afferent responses. PMID- 17400588 TI - Component information is preserved in glomerular responses to binary odor mixtures in the moth Spodoptera littoralis. AB - Natural odors are often complex mixtures of different compounds. These mixtures can be perceived to have qualities that are different from their components. Moreover, components can be difficult to distinguish within a blend, even if those components are identifiable when presented individually. Thus, odor components can interact along the olfactory pathway in a nonlinear fashion such that the mixture is not perceived simply as the sum of its components. Here we investigated odor-evoked changes in Ca2+ concentration to binary blends of plant related substances in individually identified glomeruli in the moth Spodoptera littoralis. We used a wide range of blend ratios and a range of concentrations below the level at which glomerular responses become saturated. We found no statistically significant cases where the mixture response was greater than both component responses at the same total concentration (synergistic interactions) and no statistically significant cases where the mixture response was less than either component presented individually (suppressive interactions). Therefore, we conclude that, for the plant mixtures studied, information of their components is preserved in the neural representations encoded at the first stage of olfactory processing in this moth species. PMID- 17400589 TI - Effects of dosing regimen on accumulation, retention and prophylactic efficacy of liposomal amphotericin B. AB - OBJECTIVES: We hypothesized that effective prophylactic treatment of fungal infections would require adequate drug penetration and retention at potential infection sites. Using a mouse model, we examined liposomal amphotericin B (L AmB) biodistribution, cell localization and retention in kidneys, lungs, liver and spleen to evaluate effective dosing regimens for prophylaxis of Candida glabrata and Candida albicans infections. METHODS: Following treatment of mice with cumulative doses of L-AmB (60-225 mg/kg), a bioassay was done to determine tissue drug concentrations 12 h to 6 weeks post-treatment. Immunohistochemical staining with anti-amphotericin B antibodies was used for cellular drug localization. Mice were treated prophylactically with 15-90 mg/kg L-AmB and challenged intravenously 1-7 days later with C. glabrata or they were given a total of 60 mg/kg as daily or intermittent dosing followed by intravenous challenge with C. albicans 3 or 6 weeks later. RESULTS: On the basis of microg/g tissue, the relative amount of drug was in the order spleen > liver > kidneys > lungs. Amphotericin B levels were maintained above the MIC for many fungi for 1 week in lungs and for as long as 6 weeks in kidneys and spleen. Drug localized in kidney tubular epithelial cells and in macrophages of liver and spleen. In prophylactic models, fungal burden was reduced by several 1000-fold or was undetectable within target tissues (kidneys, spleen). CONCLUSIONS: These observations underscore the importance of including drug tissue levels to obtain a better understanding of L-AmB efficacy. The sustained concentrations of bioactive AmB in many tissues provide a further rationale for investigating L-AmB prophylactic regimens. PMID- 17400590 TI - Chiari malformation and sleep related breathing disorders. AB - OBJECTIVE: To estimate the frequency, mechanisms and predictive factors of sleep apnoea syndrome (SAS) in a large group of children and adults with type I (CMI) and II (CMII) Chiari malformation (CM). BACKGROUND: The anatomical and functional integrity of both respiratory circuits and lower cranial nerves controlling the upper airway is necessary for breathing control during sleep. These latter structures may be altered in CM, and a few investigations have reported CM related sleep disordered breathing. METHODS: Forty-six consecutive unrelated patients with CM (40 CMI, six CMII), of which 20 were children (eight males) and 26 were adults (12 males), underwent physical, neurological and oto-rhino laryngoscopic examination, MRI and polysomnography. RESULTS: SAS was present in 31 (67.4%) of the patients with CM (70% of CMI, 50% of CMII, including mainly children). Sixty per cent of children with CM exhibited SAS, including 35% with obstructive (OSAS) and 25% with central (CSAS) sleep apnoea syndrome. SAS was observed in 73% of CM adults (57.7% OSAS, 15.4% CSAS). Severe SAS was found in 23% of CM adults. Multiple regression analysis revealed that age, type II Chiari and vocal cord paralysis predicted the central apnoea index. CONCLUSION: SAS is highly prevalent in all age groups of patients suffering from CM. CSAS, a rare condition in the general population, was common among the patients with CM in our study. Sleep disordered breathing associated with CM may explain the high frequency of respiratory failures observed during curative surgery of CM. Our results suggest that SAS should be systematically screened for in patients with CM, especially before surgery. PMID- 17400591 TI - Estimated life expectancy of Parkinson's patients compared with the UK population. AB - OBJECTIVE: To calculate the best possible estimates for age specific life expectancy (LE) and anticipated age at the time of death (AAD) in patients with Parkinson's disease (PD) compared with the general population in the UK. These may be of greater value to patients than standardised mortality ratios (SMRs), which are usually reported in studies on mortality in PD. METHODS: A literature review identified articles with data on age stratified life expectancy or SMRs to calculate estimations of LE using the Gompertz function and data on mortality and LE in the UK from the Office of National Statistics and Actuarial Department for the year 2003. RESULTS: Two UK studies and four from Western Europe were used to estimate LE and AAD for patients with PD from SMRs. The mean LEs of patients with PD compared with the general population were: 38 (SD 5) years for onset between 25 and 39 years compared with 49 (SD 5) years; 21 (SD 5) years for onset between 40 and 64 years compared with 31 (SD 7) years; and 5 (SD 4) years for onset age > or = 65 years compared with 9 (SD 5) years. The average AAD of patients with PD with onset between 25 and 39 years was 71 (SD 3) years and considerably lower than that of the general population (82 (SD 2) years). The difference between average AAD for older individuals with PD (onset > or = 65 years) and the general population was smaller, with an AAD of approximately 88 (SD 7) years compared with 91 (SD 5) years. CONCLUSIONS: The calculations showed that LE and AAD in PD are reduced for all onset ages but this reduction is greatest in individuals with a young onset. While the results are average estimates, these can provide useful indications of LE and AAD. PMID- 17400592 TI - Prevalence and pattern of perceived intelligibility changes in Parkinson's disease. AB - BACKGROUND: Changes to spoken communication are inevitable in Parkinson's disease (PD). It remains unclear what consequences changes have for intelligibility of speech. AIMS: To establish the prevalence of impaired speech intelligibility in people with PD and the relationship of intelligibility decline to indicators of disease progression. METHODS: 125 speakers with PD and age matched unaffected controls completed a diagnostic intelligibility test and described how to carry out a common daily activity in an "off drug" state. Listeners unfamiliar with dysarthric speech evaluated responses. RESULTS: 69.6% (n = 87) of people with PD fell below the control mean of unaffected speakers (n = 40), 51.2% (n = 64) by more than -1 SD below. 48% (n = 60) were perceived as worse than the lowest unaffected speaker for how disordered speech sounded. 38% (n = 47) placed speech changes among their top four concerns regarding their PD. Intelligibility level did not correlate significantly with age or disease duration and only weakly with stage and severity of PD. There were no significant differences between participants with tremor dominant versus postural instability/gait disorder motor phenotypes of PD. CONCLUSIONS: Speech intelligibility is significantly reduced in PD; it can be among the main concerns of people with PD, but it is not dependent on disease severity, duration or motor phenotype. Patients' own perceptions of the extent of change do not necessarily reflect objective measures. PMID- 17400593 TI - Appreciation of the informed consent procedure in a randomised trial of decompressive surgery for space occupying hemispheric infarction. AB - BACKGROUND AND AIM: As non-randomised studies have suggested that surgical decompression may reduce mortality in patients with space occupying hemispheric infarction, randomisation may be considered unethical in controlled trials testing this treatment strategy. We studied differences in recall of information and in appreciation of the informed consent procedure between representatives included in the Hemicraniectomy After Middle cerebral artery infarction with Life threatening Edema Trial (HAMLET) and representatives of patients participating in the randomised trial of Paracetamol (Acetaminophen) In Stroke (PAIS). METHODS: 1 year after study inclusion, we contacted 30 consecutive representatives who had given informed consent for participation of their relative in HAMLET, and 30 for PAIS. Recall of trial details and appreciation of the informed consent procedure were investigated using standardised questionnaires and compared between the two groups. RESULTS: All 30 PAIS representatives and 28 HAMLET representatives were interviewed. Participation of their relative in a clinical trial was remembered by 86% of HAMLET and 40% of PAIS representatives (p<0.001). HAMLET representatives remembered more trial details (effect of the treatment under study (61% vs 3%, p<0.001); randomised treatment allocation (71% vs 0%, p<0.001)). With respect to appreciation of the informed consent procedure, we found no differences between the groups: in each trial, four representatives (14% vs 13%) had considered the question of randomisation unacceptable. CONCLUSIONS: Participation of patients in a randomised controlled trial of surgical decompression for space occupying infarction is generally considered acceptable by their representatives, and recall of trial details is better than in a trial in which less vital issues are at stake. PMID- 17400594 TI - Cryopreservation of intact human ovary with its vascular pedicle--or cryopreservation of hemiovaries? PMID- 17400596 TI - Low adherence to colonoscopy in the screening of first-degree relatives of patients with colorectal cancer. AB - BACKGROUND: Colonoscopy is one of the methods of choice for screening relatives of patients with colorectal cancer. OBJECTIVE: To evaluate the rate of adherence to colonoscopy in first-degree relatives of patients with colorectal cancer and describe the lesions found. METHODS: A prospective, cross-sectional, multicentre, nationwide study was conducted. The study population was composed of first-degree relatives of patients with colorectal cancer selected randomly from the EPICOLON study. Seventy-four index patients were included. These had 342 living first degree relatives (parents, siblings and children), of whom 281 were interviewed. RESULTS: The adherence rate was 38% (107/281). Adherence was greater in families with a higher degree of familial aggregation for colorectal cancer (88.9% for Amsterdam vs 33.3% for Bethesda and sporadic cancer; p<0.05), an index patient aged under 65 years (60% for patients <65 years vs 32.9% for patients >or=65 years; p<0.05) and an index patient who was female (46.2% for women vs 31% for men; p = 0.28). Adherence was also greater in relatives under 65 years (54% in patients <65 years vs 18% in patients >or=65 years; p = 0.05), in female relatives (49% in female relatives vs 27.3% in male relatives; p<0.05) and in siblings and children (40% in siblings and children vs 13% in parents; p<0.05). Lesions were found in 26% (28/107) of the study population. Nine (8.4%) individuals had a total of 18 advanced lesions. CONCLUSIONS: These results indicate that adherence to colonoscopy in our population of first-degree relatives was low. The adherence was more frequently associated with a higher degree of familial aggregation, a relative age of under 65 years, a sibling or offspring relationship, and female sex. PMID- 17400597 TI - A temporal hidden Markov regression model for the analysis of gene regulatory networks. AB - We propose a novel hierarchical hidden Markov regression model for determining gene regulatory networks from genomic sequence and temporally collected gene expression microarray data. The statistical challenge is to simultaneously determine the groupings of genes and subsets of motifs involved in their regulation, when the groupings may vary over time, and a large number of potential regulators are available. We devise a hybrid Monte Carlo methodology to estimate parameters under 2 classes of latent structure, one arising due to the unobservable state identity of genes and the other due to the unknown set of covariates influencing the response within a state. The effectiveness of this method is demonstrated through a simulation study and an application on an yeast cell-cycle data set. PMID- 17400599 TI - Microglial activation in presymptomatic Huntington's disease gene carriers. AB - Microglial activation may play a role in the pathogenesis of Huntington's disease (HD). Using 11C-(R)-PK11195 (PK) positron emission tomography (PET), we investigated microglial activation in HD presymptomatic gene carriers (PGCs), its relationship with striatal neuronal dysfunction measured with 11C-raclopride (RAC) PET, and the role of PK PET as a possible marker of subclinical disease progression in PGCs. Eleven HD PGCs underwent PK and RAC PET. Their results were compared with those of healthy controls. PK and RAC binding was measured using region-of-interest analysis. Regional increases in PK binding were also localized with voxel-based statistical parametric mapping. HD PGCs had lower striatal RAC binding than the controls but significantly higher striatal and cortical PK binding. Individual levels of higher striatal PK binding in PGCs correlated with lower striatal RAC binding and, after excluding one outlier, with a higher probability of developing HD in 5 years. The inverse association between striatal PK and RAC binding in PGCs continues into early to moderate stages of HD. This study demonstrated for the first time in vivo widespread microglial activation in preclinical HD which correlated with striatal neuronal dysfunction. These findings indicate that microglial activation is an early event in the pathogenic processes of HD and is associated with subclinical progression of disease. PK PET may be a useful marker of active subclinical disease and a means of investigating the efficacy of neuroprotection strategies in PGCs. PMID- 17400600 TI - Developmental differences in the responses of IL-6 and IL-13 transgenic mice exposed to hyperoxia. AB - Our previous work has shown that adult mice with overexpression of IL-6 and IL-13 in the lung have enhanced survival in hyperoxia associated with reduced hyperoxia induced lung injury and cell death. We hypothesized that there are developmental differences in these responses in the adult vs. the newborn (NB) animal, and these responses have clinical relevance in the human NB. We compared the responses to 100% O(2) of NB IL-6 and IL-13 transgenic mice with wild-type littermate controls by evaluating mortality, lung tissue TUNEL staining, and mRNA expression using RT-PCR. We used ELISA to measure IL-6 levels in tracheal aspirates from human neonates. Our results show that, in contrast to the cytoprotective effects in mature mice, IL-6 caused significantly increased mortality, DNA injury, caspases, cell death regulator and angiogenic factor expression in hyperoxia in the NB. Furthermore, tracheal aspirate levels of IL-6 were significantly increased in premature neonates with respiratory distress syndrome who had an adverse outcome (bronchopulmonary dysplasia/death). In contrast to the protective effects in adults, there was no survival advantage to the NB IL-13 mice in hyperoxia. These findings imply that caution should be exercised in extrapolating results from the adult to the NB. PMID- 17400601 TI - TGF-beta-neutralizing antibodies improve pulmonary alveologenesis and vasculogenesis in the injured newborn lung. AB - Pulmonary injury is associated with the disruption of alveologenesis in the developing lung and causes bronchopulmonary dysplasia (BPD) in prematurely born infants. Transforming growth factor (TGF)-beta is an important regulator of cellular differentiation and early lung development, and its levels are increased in newborn lung injury. Although overexpression of TGF-beta in the lungs of newborn animals causes pathological features that are consistent with BPD, the role of endogenous TGF-beta in the inhibition of the terminal stage of lung development is incompletely understood. In this investigation, the hypothesis that O(2)-induced injury of the maturing lung is associated with TGF-beta mediated disruption of alveologenesis and microvascular development was tested using a murine model of BPD. Here we report that treatment of developing mouse lungs with TGF-beta-neutralizing antibodies attenuates the increase in pulmonary cell phospho-Smad2 nuclear localization, which is indicative of augmented TGF beta signaling, associated with pulmonary injury induced by chronic inhalation of 85% oxygen. Importantly, the neutralization of the abnormal TGF-beta activity improves quantitative morphometric indicators of alveologenesis, extracellular matrix assembly, and microvascular development in the injured developing lung. Furthermore, exposure to anti-TGF-beta antibodies is associated with improved somatic growth in hyperoxic mouse pups and not with an increase in pulmonary inflammation. These studies indicate that excessive pulmonary TGF-beta signaling in the injured newborn lung has an important role in the disruption of the terminal stage of lung development. In addition, they suggest that anti-TGF-beta antibodies may be an effective therapy for preventing some important developmental diseases of the newborn lung. PMID- 17400602 TI - Brief report: sleep in children with sickle cell disease: an analysis of daily diaries utilizing multilevel models. AB - OBJECTIVE: To investigate the pain-sleep relationship in children with sickle cell disease (SCD) and the influence of stress and pain medication use on this relationship. METHOD: Children with SCD (n = 20; aged 8-12 years) completed daily diaries assessing sleep, pain, stress, and pain medication use for up to 2 months. Data analyzed using multilevel modeling. RESULTS: High daily pain was related to poor sleep quality that night and poor sleep quality was related to high pain the following day. High stress was related to less sleep. High same-day pain and pain medication attenuated the impact of pain on sleep quality. CONCLUSION: Results highlight the importance of sleep in addressing functioning in children with chronic pain, knowledge which may help patients and their families better manage the child's pain. Behavioral pain interventions may be improved by the inclusion of strategies to encourage proper sleep hygiene and address sleep issues. PMID- 17400603 TI - Analysis of rhythm variation during spontaneous cardioinhibitory neurally mediated syncope. Implications for RDR pacing optimization: an ISSUE 2 substudy. AB - BACKGROUND: Little is known of the variations of the heart rate during spontaneous cardioinhibitory neurally-mediated syncope. Their knowledge has both academic and practical implications for the optimization of rate drop response (RDR) pacing mode. METHODS AND RESULTS: We describe variations of the rhythm occurring during 48 syncopal episodes documented by implantable loop recorder. The presyncopal phase of 18 s (interquartile range 9-65) was characterized by a fall in heart rate from 83 +/- 20 bpm to maximal bradycardia or (multiple) asystolic pauses which lasted a median of 19 s (10-30). The recovery phase lasted 22 s (7-52). The total duration of the cardioinhibitory reflex was 85 s (47-116). We then calculated the potential increase in benefit that an optimally programmed drop rate detection could provide compared with a reference Lower Rate detection. Compared with Lower Rate detection (defined as two consecutive beats at 40 bpm), drop rate detection (assumed to be drop size = 20 bpm, detection window = 1 min, and drop rate = 50 bpm) would have been able to introduce intervention pacing, a median of 5.7 s (interquartile range -5.1- -10.4) earlier in 28 cases (58%). CONCLUSION: Cardioinhibitory neurally-mediated reflex varies widely from a few seconds to some minutes. In our data the total duration was <2 min. Optimal RDR programming, being potentially able to anticipate the detection of the cardioinhibitory reflex by a few seconds, could provide an increase in benefit for cardiac pacing therapy in prevention of syncope. PMID- 17400604 TI - Pacing-related acute pulmonary edema and mechanical asynchrony illustrated by three-dimensional echocardiography. AB - Sick sinus syndrome with symptomatic bradycardia is an indication for a permanent pacemaker. Either a single (AAIR) or dual-chamber (DDDR) pacemaker can be implanted in these patients with normal atrioventricular nodal function. This report presents a 92-year-old male with right ventricular apical pacing related recurrent acute pulmonary edema and mechanical asynchrony demonstrated by three dimensional echocardiogram. Although three-dimensional echocardiography has been available for many years, it has seldom been applied to evaluate pacing-related intraventricular asynchrony. The systolic asynchrony index for this patient was 6.7% during AAIR pacing mode and 22% during DDDR pacing mode. PMID- 17400605 TI - Fibrinolysis may widen the time window for primary angioplasty. PMID- 17400606 TI - Heme oxygenase-1 gene promoter polymorphism and restenosis following coronary stenting. AB - AIMS: Gene expression analyses, cell culture experiments, animal models, and association studies suggest a protective role of the heme oxygenase-1 (HO-1) protein against restenosis. The length of a polymorphic (GT)n dinucleotid repeats sequence in the HO-1 gene promoter influences the transcriptional activity. We evaluated, whether an association existed between this polymorphism and the incidence of restenosis after coronary stenting. METHODS AND RESULTS: Of the 1807 consecutive patients included in this study, 1357 (75%) patients had 6 months follow-up angiography. Restenosis, the primary endpoint, was defined as angiographic restenosis, diameter stenosis of > or =50%, and clinical restenosis, target vessel revascularization during the first year. The combined 1 year incidence of death and myocardial infarction (MI) was evaluated as secondary endpoint. We divided the alleles similar to previous studies: class S less repeats (<25), and class L more repeats (> or =25), leading to SS, SL, and LL genotypes. Angiographic restenosis rate showed no significant difference for the studied genotypes-SS 29.2%, SL 29.5%, and LL genotype 29.6% (P = 0.99). There was no significant difference regarding clinical restenosis (P = 0.28) and combined incidence of death or MI (P = 0.98). CONCLUSION: This study does not support a clinically relevant association of the HO-1 promoter polymorphism with restenosis and ischaemic events after coronary stenting. PMID- 17400607 TI - Metabolic syndrome: nothing more than a constellation? PMID- 17400608 TI - Effect of dynamic myocardial dyssynchrony on mitral regurgitation during supine bicycle exercise stress echocardiography in patients with idiopathic dilated cardiomyopathy and 'narrow' QRS. AB - AIMS: Cardiac resynchronization therapy (CRT) has become an attractive therapeutic option for patients with end-stage heart failure (HF). Currently, patients are selected for CRT on ECG and on echocardiographic criteria analysed at rest. Whether the physical effort may further increase myocardial dyssynchrony is not fully established. The aim of the study was to test by the use of Doppler myocardial imaging (DMI) if dynamic left ventricular (LV) dyssynchrony during physical effort may be a determinant of dynamic mitral regurgitation in patients with dilated cardiomyopathy and 'narrow' QRS. METHODS AND RESULTS: Sixty patients (62.3 +/- 8.3 years) with idiopathic dilated cardiomyopathy and narrow QRS duration ( < 120 ms) were selected. All the patients underwent standard Doppler echo, colour DMI, supine bicycle exercise stress echocardiography, and cardiopulmonary exercise testing. Cardiac synchronicity was assessed, at rest and at peak exercise, from measurements of time intervals (Ts) between the onset of the QRS complex and the peak myocardial systolic velocity, in a six-basal-six-mid segmental model. Standard deviation of Ts of the 12 LV segments (Ts-SD-12) was also calculated. In baseline conditions, HF patients showed an LV ejection fraction of 30.1 +/- 4%, and a significant electromechanical delay (Ts-SD-12 > or = 34.4 ms) in 20 patients (33.3%). At peak of physical exercise, a significant electromechanical delay was detected in 35 patients (58.3%), whereas in 47 patients (78.3%) exercise-induced increase in mitral valve effective regurgitant orifice (ERO) was observed. By multivariable analysis, an independent positive association between changes in Ts-SD-12 and in mitral valve ERO (P < 0.0001), as well as an independent inverse correlation of the same changes in Ts-SD-12 with LV stroke volume (P < 0.0001) were detected. In addition, changes in Ts-SD-12 were also independent determinants of peak VO(2) (P < 0.0001) during cardiopulmonary exercise testing. CONCLUSION: Colour DMI is an effective technique for assessing the severity of regional delay in activation of LV walls in HF patients with narrow QRS both at rest and during stress test. The increase in LV dyssynchrony during exercise strongly correlates with the increase in mitral regurgitation severity and with the impairment of LV stroke volume. PMID- 17400609 TI - Localization of hCAP-18 on the surface of chemoattractant-stimulated human granulocytes: analysis using two novel hCAP-18-specific monoclonal antibodies. AB - The well-described antimicrobial and immunoregulatory properties of human cathelicidin antimicrobial protein 18 (hCAP-18) derive in part from the ability of its proteolytic fragment, LL-37 (a.k.a. CAP-37), to associate with activated immune and epithelial cells during inflammation. We now show a stable association between hCAP-18 and the cell surface of formyl-Met-Leu-Phe (fMLF)-stimulated neutrophils using two novel hCAP-18-specific mAb, H7 and N9, which recognize a single 16-kDa band, identified by N-terminal sequencing and mass spectrometry as hCAP-18. Phage display analysis of epitope-binding sites showed that both mAb probably recognize a similar five amino acid sequence near the C terminus of the prodomain. Immunoblot analysis of degranulated neutrophil supernatants resulted in mAb recognition of the 14-kDa prodomain of hCAP-18. Subcellular fractionation of unstimulated neutrophils on density gradients showed expected cosedimentation of hCAP-18 with specific granule lactoferrin (LF). fMLF stimulation resulted in an average 25% release of specific granule hCAP-18, with approximately 15% of the total cellular hCAP-18 recovered from culture media, and approximately 10% and approximately 75%, respectively, codistributing with plasma membrane alkaline phosphatase and specific granule LF. Surface association of hCAP-18 on fMLF stimulated neutrophils was confirmed by immunofluorescence microscopy and flow cytometry analysis, which also suggested a significant up-regulation of surface hCAP-18 on cytochalasin B-pretreated, fully degranulated neutrophils. hCAP-18 surface association was labile to 10 mM NaOH treatment but resistant to 1 M NaCl and also partitioned into the detergent phase following Triton X-114 solubilization, possibly suggesting a stable association with one or more integral membrane proteins. We conclude that fMLF stimulation promotes redistribution of hCAP-18 to the surface of human neutrophils. PMID- 17400610 TI - Protection of CD8+ T cells from activation-induced cell death by IL-18. AB - Role of IL-18 on proliferation and survival of CD8+ T cells, activated by immobilized anti-CD3 antibody (anti-CD3), was examined. Proliferation and survival of activated T cells, especially that of CD8+ T cells, were impaired by IL-18 deficiency [IL-18 knockout (KO)]. After 3 days of culture with anti-CD3, the number of living CD8+ T cells from IL-18KO mice was approximately 25% of that from wild-type (WT) mice but was increased to the same level as WT cells by the addition of IL-18. The expression of IL-18 receptors (IL-18Rs), particularly IL 18Rbeta chain, in naive CD8+ T cells was very low but elevated after stimulation with anti-CD3. Blockade of IL-18R by anti-IL-18R antibody on activated WT CD8+ T cells resulted in reduction of living cells, suggesting that IL-18 promotes survival of proliferating CD8+ T cells. Levels of Bcl-2 in activated IL-18KO CD8+ T cells were lower than those in WT cells but were raised by exogenous IL-18. Blockade of IL-18R on WT CD8+ T cells decreased the expression of surface markers CD122 and CD94, which are related to cell viability, and the expression of these markers was increased by exogenous IL-18 in IL-18KO cells. These results suggest that IL-18 acts directly on activated CD8+ T cells through IL-18Rs and promotes their survival to expand the population. PMID- 17400611 TI - Rapid transit in the immune cells: the role of mRNA turnover regulation. AB - There have been recent, significant advances about the role of mRNA turnover in controlling gene expression in immune cells. Post-transcriptional regulation of gene expression contributes to the characteristics of many of the processes underlying the immune response by ensuring early, rapid, and transient action. The emphasis of this review is on current work that deals with the regulation of mRNA decay during innate immunity against microbes and T cell activation as a model of the adaptive response. PMID- 17400612 TI - Terminal sedation: source of a restless ethical debate. PMID- 17400613 TI - Financial incentives for antipsychotic depot medication: ethical issues. AB - BACKGROUND: Giving money as a direct incentive for patients in exchange for depot medication has proved beneficial in some clinical cases in assertive outreach (AO). However, ethical concerns around this practice have been raised, and will be analysed in more detail here. METHOD: Ethical concern voiced in a survey of all AO teams in England were analysed regarding their content. These were grouped into categories. RESULTS: 53 of 70 team managers mentioned concerns, many of them serious and expressing a negative attitude towards giving money for depot adherence. Four broad categories of ethical concern following Christensen's concept were distinguished: valid consent and refusal (n = 5), psychiatric paternalism (n = 31), resource allocation (n = 4), organisational relationships (n = 2), with a residual category others and unspecified (n = 11). DISCUSSION: The main concerns identified are discussed on the background of existing ethical theories in healthcare and the specific problems of community mental health and AO. Points for practice are derived from this discussion. A way forward is outlined that includes informed consent and an operational policy in the use of incentives, further randomised controlled trials and qualitative studies, and continuing discussions with all stakeholders, especially service users. PMID- 17400614 TI - "Do-not-resuscitate" orders in patients with cancer at a children's hospital in Taiwan. AB - OBJECTIVES: To quantify the use of do-not-resuscitate (DNR) orders in a tertiary care children's hospital and to characterise the circumstances in which such orders are written. DESIGN: Retrospective study conducted in a 500-bed children's hospital in Taiwan. PATIENTS: The course of 101 patients who died between January 2002 and December 2005 was reviewed. The following data were collected: age at death, gender, disease and its status, place of death and survival. There were 59 males and 42 females with a median age of 103 months (range 1-263 months). 50 children had leukaemias, and 51 had malignancies other than leukaemia. The t test and the chi(2) test were applied as appropriate. RESULTS: The study found that 44% of patient deaths occurred in the paediatric oncology ward; 29% of patient deaths occurred in the intensive care unit; and 28% of patients died in their home or at another hospital. Other findings included the following: 46 of 101 (46%) patients died after attempted cardiopulmonary resuscitation and 55 (54%) died with a DNR order in effect. The mean age at death was 9.8 years in both groups with or without DNR orders. CONCLUSIONS: From the study of patient deaths in this tertiary-care children's hospital, it was concluded that an explicit DNR order is now the rule rather than the exception, with more DNR orders being written for patients who have been ill longer, who have solid tumours, who are not in remission and who are in the ward. PMID- 17400616 TI - A "Queen of Hearts" trial of organ markets: why Scheper-Hughes's objections to markets in human organs fail. AB - Nancy Scheper-Hughes is one of the most prominent critics of markets in human organs. Unfortunately, Scheper-Hughes rejects the view that markets should be used to solve the current (and chronic) shortage of transplant organs without engaging with the arguments in favour of them. Scheper-Hughes's rejection of such markets is of especial concern, given her influence over their future, for she holds, among other positions, the status of an adviser to the World Health Organization (Geneva) on issues related to global transplantation. Given her influence, it is important that Scheper-Hughes's moral condemnation of markets in human organs be subject to critical assessment. Such critical assessment, however, has not generally been forthcoming. A careful examination of Scheper Hughes's anti-market stance shows that it is based on serious mischaracterisations of both the pro-market position and the medical and economic realities that underlie it. In this paper, the author will expose and correct these mischaracterisations and, in so doing, show that her objections to markets in human organs are unfounded. PMID- 17400615 TI - The concept of brain death did not evolve to benefit organ transplants. AB - Although it is commonly believed that the concept of brain death (BD) was developed to benefit organ transplants, it evolved independently. Transplantation owed its development to advances in surgery and immunosuppressive treatment; BD owed its origin to the development of intensive care. The first autotransplant was achieved in the early 1900s, when studies of increased intracranial pressure causing respiratory arrest with preserved heartbeat were reported. Between 1902 and 1950, the BD concept was supported by the discovery of EEG, Crile's definition of death, the use of EEG to demonstrate abolition of brain potentials after ischaemia, and Crafoord's statement that death was due to cessation of blood flow. Transplantation saw the first xenotransplant in humans and the first unsuccessful kidney transplant from a cadaver. In the 1950s, circulatory arrest in coma was identified by angiography, and the death of the nervous system and coma depasse were described. Murray performed the first successful kidney transplant. In the 1960s, the BD concept and organ transplants were instantly linked when the first kidney transplant using a brain-dead donor was performed; Schwab proposed to use EEG in BD; the Harvard Committee report and the Sydney Declaration appeared; the first successful kidney, lung and pancreas transplants using cadaveric (not brain-dead) donors were achieved; Barnard performed the first human heart transplant. This historical review demonstrates that the BD concept and organ transplantation arose separately and advanced in parallel, and only began to progress together in the late 1960s. Therefore, the BD concept did not evolve to benefit transplantation. PMID- 17400617 TI - Benefit sharing: it's time for a definition. AB - Benefit sharing has been a recurrent theme in international debates for the past two decades. However, despite its prominence in law, medical ethics and political philosophy, the concept has never been satisfactorily defined. In this conceptual paper, a definition that combines current legal guidelines with input from ethics debates is developed. Philosophers like boxes; protective casings into which they can put concisely-defined concepts. Autonomy is the human capacity for self determination; beneficence denotes the virtue of good deeds, coercion is the intentional threat of harm and so on. What about benefit sharing? Does the concept have a box and are the contents clearly defined? The answer to this question has to be no. The concept of benefit sharing is almost unique in that various disciplines use it regularly without precise definitions. In this article, a definition for benefit sharing is provided, to eliminate unnecessary ambiguity. PMID- 17400618 TI - The unhealthy physician. AB - BACKGROUND: Physicians, if affected by transmissible or impairing diseases, could be hazardous for third persons. AIM: To solve the apparent chasm between patient's and sick worker's rights, a consensus-building process leading to hospital-wide policies is the better alternative to individual decision making. CONCLUSIONS: Policies have to balance the rights of the sick worker, the right of the other workers, patients and customers, and society's expectations. PMID- 17400619 TI - Withholding and withdrawing life support in critical care settings: ethical issues concerning consent. AB - The right to refuse medical intervention is well established, but it remains unclear how best to respect and exercise this right in life support. Contemporary ethical guidelines for critical care give ambiguous advice, largely because they focus on the moral equivalence of withdrawing and withholding care without confronting the very real differences regarding who is aware and informed of intervention options and how patient values are communicated and enacted. In withholding care, doctors typically withhold information about interventions judged too futile to offer. They thus retain greater decision-making burden (and power) and face weaker obligations to secure consent from patients or proxies. In withdrawing care, there is a clearer imperative for the doctor to include patients (or proxies) in decisions, share information and secure consent, even when continued life support is deemed futile. How decisions to withhold and withdraw life support differ ethically in their implications for positive versus negative interpretations of patient autonomy, imperatives for consent, definitions of futility and the subjective evaluation of (and submission to) benefits and burdens of life support in critical care settings are explored. Professional reflection is required to respond to trends favouring a more positive interpretation of patient autonomy in the context of life support decisions in critical care. Both the bioethics and critical care communities should investigate the possibilities and limits of growing pressure for doctors to disclose their reasoning or seek patient consent when decisions to withhold life support are made. PMID- 17400620 TI - Transplants save lives, defending the double veto does not: a reply to Wilkinson. PMID- 17400621 TI - How to take deontological concerns seriously in risk-cost-benefit analysis: a re interpretation of the precautionary principle. AB - In this paper the coherence of the precautionary principle as a guide to public health policy is considered. Two conditions that any account of the principle must meet are outlined, a condition of practicality and a condition of publicity. The principle is interpreted in terms of a tripartite division of the outcomes of action (good outcomes, normal bad outcomes and special bad outcomes). Such a division of outcomes can be justified on either "consequentialist" or "deontological" grounds. In the second half of the paper, it is argued that the precautionary principle is not necessarily opposed to risk-cost-benefit analysis, but, rather, should be interpreted as suggesting a lowering of our epistemic standards for assessing evidence that there is a link between some policy and "special bad" outcomes. This suggestion is defended against the claim that it mistakes the nature of statistical testing and against the charge that it is unscientific or antiscientific, and therefore irrational. PMID- 17400622 TI - Concepts of "person" and "liberty," and their implications to our fading notions of autonomy. AB - It is commonly held that respect for autonomy is one of the most important principles in medical ethics. However, there are a number of interpretations as to what that respect actually entails in practice and a number of constraints have been suggested even on our self-regarding choices. These limits are often justified in the name of autonomy. In this paper, it is argued that these different interpretations can be explained and understood by looking at the discussion from the viewpoints of positive and negative liberty and the various notions of a "person" that lay beneath. It will be shown how all the appeals to positive liberty presuppose a particular value system and are therefore problematic in multicultural societies. PMID- 17400623 TI - Consent gained from patients after breast surgery for the use of surplus tissue in research: an exploration. AB - OBJECTIVES: (1) To investigate the quality of consent gained for the use in research of tissue that is surplus after surgery. (2) To compare the use of two consent forms: a simple locally introduced form and a more complex centrally instigated form. (3) To discuss the attitudes of patients towards the use of their surplus tissue in research. DESIGN: Data were collected through interviews and analysed with a combination of quantitative and qualitative analytical techniques. PARTICIPANTS AND SETTING: Patients of the breast care unit at a teaching hospital were interviewed at home or in a quiet room at the hospital. RESULTS: 57 people were interviewed out of 81 approached, between October 2003 and March 2004. Most participants had a poor level of knowledge about the consent they had given, but reported being happy about having given it. The patients who had signed the locally introduced form had considerably more knowledge than those who had signed the centrally instigated form (z = -2.56; p<0.05). Participants considered being well informed to be less important than believing that their opinions were valued and respected. CONCLUSIONS: The findings suggest that traditional models of informed consent are not universally applicable and, in this case, seem to overstate what people wish to know. The simple consent form achieved a better quality of informed consent and provided a better model of practice than the complex form, and it seemed that a focused approach to consent seeking is more effective and acceptable than more complex approaches. PMID- 17400624 TI - Attitudes towards clinical research among cancer trial participants and non participants: an interview study using a Grounded Theory approach. AB - The attitudes of women patients with cancer were explored when they were invited to participate in one of three randomised trials that included chemotherapy at two university centres and a satellite centre. Fourteen patients participating in and 15 patients declining trials were interviewed. Analysis was based on the constant comparative method. Most patients voiced positive attitudes towards clinical research, believing that trials are necessary for further medical development, and most spontaneously argued that participation is a moral obligation. Most trial decliners, however, described a radical change in focus as they faced the actual personal choice. Almost no one got an impression of clinical equipoise between treatments in the trials, and most patients expressed discomfort with randomisation. A patient's choice to participate was mainly determined by whether the primary focus was on treatment effect or on adverse effects. Both knowledge about and feelings towards trials originated mostly from the media, although paradoxically the media were largely seen as untrustworthy. Mistrust was shown towards the pharmaceutical industry, and although most patients originally trusted that doctors primarily pursued the interest of patients, they did not trust the adequacy of doctors or industry in maintaining self-regulation. Thus, public control measures were judged to be essential. PMID- 17400625 TI - Proportional ethical review and the identification of ethical issues. AB - Presently, there is a movement in the UK research governance framework towards what is referred to as proportional ethical review. Proportional ethical review is the notion that the level of ethical review and scrutiny given to a research project ought to reflect the level of ethical risk represented by that project. Relatively innocuous research should receive relatively minimal review and relatively risky research should receive intense scrutiny. Although conceptually attractive, the notion of proportional review depends on the possibility of effectively identifying the risks and ethical issues posed by an application with some process other than a full review by a properly constituted research ethics committee. In this paper, it is argued that this cannot be achieved and that the only appropriate means of identifying risks and ethical issues is consideration by a full committee. This implies that the suggested changes to the National Health Service research ethics system presently being consulted on should be strenuously resisted. PMID- 17400626 TI - Risk in public health and clinical work. PMID- 17400628 TI - Teaching tip: using the "Who wants to be a millionaire?" game to teach searching skills. PMID- 17400629 TI - Physiotherapy for tennis elbow. PMID- 17400630 TI - Methods in research: key principles of graph construction. PMID- 17400631 TI - Corticosteroid injections or physiotherapy were not more effective than wait and see for tennis elbow at 1 year. PMID- 17400632 TI - Review: evidence from 2 low quality screening studies does not show a reduction in death from prostate cancer. PMID- 17400633 TI - Vaccinating infants with a wide long needle was non-inferior to a narrow short needle for immune response. PMID- 17400635 TI - Atorvastatin at 80 mg/day reduced cerebrovascular events more than atorvastatin at 10 mg/day in stable coronary heart disease. PMID- 17400634 TI - Review: high dose statins reduce risk of non-fatal cardiovascular events more than standard dose statins. PMID- 17400636 TI - Physical counterpressure manoeuvres reduced vasovagal syncope. PMID- 17400637 TI - An abnormal D-dimer test result indicated that anticoagulant therapy should be continued. PMID- 17400638 TI - Use of a 2 hour partogram action line instead of a 4 hour action line did not reduce caesarean delivery rate. PMID- 17400639 TI - Review: children <2 years of age with bilateral acute otitis media and children with otorrhoea benefit most from antibiotics. PMID- 17400640 TI - The Air-Stirrup brace with elastic wrap improved function in first incident grade 1 or 2 lateral ankle ligament sprain. PMID- 17400641 TI - Addition of peak flow monitoring to symptom monitoring did not improve healthcare visits, quality of life, or lung function in older adults with moderate-to-severe asthma. PMID- 17400642 TI - A lifestyle intervention continued to prevent type 2 diabetes in high risk patients after the programme was stopped. PMID- 17400644 TI - Review: inhaled insulin provided better glycaemic control than oral hypoglycaemic agents but not subcutaneous insulin. PMID- 17400643 TI - Community based occupational therapy improved daily functioning in older patients with dementia. PMID- 17400645 TI - Review: pioglitazone does not reduce risk of mortality or cardiovascular events in type 2 diabetes. PMID- 17400646 TI - Clinical examination had poor sensitivity for detecting non-cephalic presentation in late pregnancy. PMID- 17400647 TI - Review: alarm features in dyspepsia do not have high diagnostic accuracy for predicting upper gastrointestinal cancer. PMID- 17400648 TI - Review: sensitivities and specificities vary for different diagnostic tests of gout. PMID- 17400649 TI - A risk model predicted major bleeding in older patients with atrial fibrillation receiving warfarin therapy. PMID- 17400650 TI - Simvastatin was cost effective across a broad range of risk and age groups. PMID- 17400652 TI - Comment on the editorial "Misunderstandings, misperceptions, and mistakes". PMID- 17400653 TI - Single nucleotide polymorphism associated with mature miR-125a alters the processing of pri-miRNA. AB - MicroRNAs (miRNAs) are small non-coding RNAs that inhibit expression of specific target genes at the post-transcriptional level. Sequence variations in miRNA genes, including pri-miRNAs, pre-miRNAs and mature miRNAs, could potentially influence the processing and/or target selection of miRNAs. In this study, we have systematically identified single nucleotide polymorphisms (SNPs) associated with 227 known human miRNAs. Among 323 total SNPs that we identify, 12 are located within the miRNA precursor and one is at the eighth nucleotide (+8) of the mature miR-125a, which has been proposed to play a critical role in recognition of mRNA targets by miRNAs. Through a series of in vivo analyses, we unexpectedly find that this miR-125a SNP significantly blocks the processing of pri-miRNA to pre-miRNA, in addition to reducing miRNA-mediated translational suppression. Thus, our study reveals an additional structural requirement for pri miRNA processing and emphasizes the importance of identifying new miRNA SNPs and their contributions to miRNA biogenesis and human genetic disease. PMID- 17400654 TI - Loss of MMP-2 disrupts skeletal and craniofacial development and results in decreased bone mineralization, joint erosion and defects in osteoblast and osteoclast growth. AB - The 'vanishing bone' or inherited osteolysis/arthritis syndromes represent a heterogeneous group of skeletal disorders characterized by mineralization defects of affected bones and joints. Differing in anatomical distribution, severity and associated syndromic features, gene identification in each 'vanishing bone' disorder should provide unique insights into genetic/molecular pathways contributing to the overall control of skeletal growth and development. We previously described and then demonstrated that the novel autosomal recessive osteolysis/arthritis syndrome, multicentric osteolysis with arthritis (MOA) (MIM #605156), was caused by inactivating mutations in the MMP2 gene [Al Aqeel, A., Al Sewairi, W., Edress, B., Gorlin, R.J., Desnick, R.J. and Martignetti, J.A. (2000) Inherited multicentric osteolysis with arthritis: A variant resembling Torg syndrome in a Saudi family. Am. J. Med. Genet., 93, 11-18.]. These in vivo results were counterintuitive and unexpected since previous in vitro studies suggested that MMP-2 overexpression and increased activity, not deficiency, would result in the bone and joint features of MOA. The apparent lack of a murine model [Itoh, T., Ikeda, T., Gomi, H., Nakao, S., Suzuki, T. and Itohara, S. (1997) Unaltered secretion of beta-amyloid precursor protein in gelatinase A (matrix metalloproteinase 2)-deficient mice. J. Biol. Chem., 272, 22389-22392.] has hindered studies on disease pathogenesis and, more fundamentally, in addressing the paradox of how functional loss of a single proteolytic enzyme results in an apparent increase in bone loss. Here, we report that Mmp2-/- mice display attenuated features of human MOA including progressive loss of bone mineral density, articular cartilage destruction and abnormal long bone and craniofacial development. Moreover, these changes are associated with markedly and developmentally restricted decreases in osteoblast and osteoclast numbers in vivo. Mmp2-/- mice have approximately 50% fewer osteoblasts and osteoclasts than control littermates at 4 days of life but these differences have nearly resolved by 4 weeks of age. In addition, despite normal cell numbers in vivo at 8 weeks of life, Mmp2-/- bone marrow cells are unable to effectively support osteoblast and osteoclast growth and differentiation in culture. Targeted inhibition of MMP-2 using siRNA in human SaOS2 and murine MC3T3 osteoblast cell lines resulted in decreased cell proliferation rates. Taken together, our findings suggest that MMP 2 plays a direct role in early skeletal development and bone cell growth and proliferation. Thus, Mmp2-/- mice provide a valuable biological resource for studying the pathophysiological mechanisms underlying the human disease and defining the in vivo physiological role of MMP-2. PMID- 17400655 TI - Neurofibromatosis-1 (Nf1) heterozygous brain microglia elaborate paracrine factors that promote Nf1-deficient astrocyte and glioma growth. AB - The tumor microenvironment is considered to play an important role in tumor formation and progression by providing both negative and positive signals that influence tumor cell growth. We and others have previously shown that brain tumor (glioma) formation in Nf1 genetically engineered mice requires a microenvironment composed of cells heterozygous for a targeted Nf1 mutation. Using NF1 as a model system to understand the contribution of the tumor microenvironment to glioma formation, we show that Nf1+/- brain microglia produce specific factors that promote Nf1-/- astrocyte growth in vitro and in vivo and identify hyaluronidase as one of these factors in both genetically engineered Nf1 mouse and human NF1 associated optic glioma. We further demonstrate that blocking hyaluronidase ameliorates the ability of Nf1+/- microglia to increase Nf1-/- astrocyte proliferation and that hyaluronidase increases Nf1-/- astrocyte proliferation in an MAPK-dependent fashion. Lastly, inhibiting microglia activation in genetically engineered Nf1 mice significantly reduces mouse optic glioma proliferation in vivo. Collectively, these studies identify Nf1+/- microglia as an important stromal cell type that promotes Nf1-/- astrocyte and optic glioma growth relevant to the pathogenesis of NF1-associated brain tumors and suggest that future brain therapies might be directed against paracrine factors produced by cells in the tumor microenvironment. PMID- 17400656 TI - Body mass index and mortality in patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: To examine the relationship between body mass index (BMI) and mortality in patients with idiopathic pulmonary fibrosis (IPF). METHODS: We studied a cohort of patients with IPF who were seen at the Mayo Clinic Rochester from 1994 through 1996. These patients met the current consensus definition of IPF. We excluded patients who had received prior treatment for IPF, had no follow up data, or had no pulmonary function results available at the index visit. RESULTS: Of the 197 patients fulfilling the inclusion criteria, the mean (+/- SD) age was 71.4 +/- 8.9 years, 137 patients (70%) were men, and the mean BMI was 28.2 +/- 4.6. These patients were categorized by BMI into the following three groups: < 25; 25 to 30; and >/= 30. There were 46 patients (23%) with a BMI of < 25 who had a median survival time of 3.6 years (1-year survival rate, 76% [95% confidence interval (CI), 65 to 90%]; 3-year survival rate, 54% [95% CI, 41 to 70%]). The second group consisted of 85 patients (43%) with a BMI between 25 and 30 who had a median survival time of 3.8 years (1-year survival rate, 84% [95% CI, 76 to 92%]; 3-year survival rate, 58% [95% CI, 48 to 70%]). The final group consisted of 66 patients (34%) with a BMI of >/= 30 and who had a median survival time of 5.8 years (1-year survival rate, 91% [95% CI, 84 to 98%]; 3-year survival rate, 69% [95% CI, 58 to 81%]). Using a proportional hazards regression model, survival was significantly associated with BMI (hazard ratio, 0.93 for each 1-U increase in BMI; 95% CI, 0.89 to 0.97; p = 0.002) with increased BMI being associated with better survival. CONCLUSION: Higher BMI was associated with better survival in patients with IPF. PMID- 17400657 TI - Transient atrial fibrillation complicating acute inferior myocardial infarction: implications for future risk of ischemic stroke. AB - BACKGROUND: Atrial fibrillation (AF) that occurs as a frequent complication of myocardial infarction (MI) is associated with a poor clinical outcome. It nonetheless remains uncertain whether AF that occurs transiently during MI is associated with a subsequent increased risk of the development of AF and ischemic stroke. METHODS: We retrospectively studied the impact of transient AF on the long-term risk of the occurrence of AF, ischemic stroke, and mortality in 431 consecutive patients (mean [+/- SEM] age, 64 +/- 1 years; 75% men). All patients had experienced an acute inferior ST-segment-elevation MI and had preserved left ventricular ejection fraction (LVEF) [> 45%]. RESULTS: All patients were in sinus rhythm on hospital admission, and transient AF was observed in 59 patients (13.7%) during their hospitalization for MI. On hospital discharge, all patients were in sinus rhythm and had been prescribed antiplatelet agents alone as antithrombotic therapy. Patients in whom transient AF developed during MI were older (mean age, 70 +/- 1.4 vs 64 +/- 0.7 years, respectively; p < 0.01) and more likely to be women (37% vs 23%, respectively; p < 0.02) compared with those without AF. At 1-year follow-up, the incidence of AF (22.0% vs 1.3%, respectively; p < 0.01) and ischemic stroke (10.2% vs 1.8%, respectively; p < 0.01) was higher in patients with transient AF than in those without transient AF. The total mortality rate was nonetheless similar (5.6% vs 6.8%, respectively; p = 0.73); Cox regression analysis demonstrated that age > 65 years and transient AF during MI were independent predictors of the subsequent occurrence of AF and the development of ischemic stroke. CONCLUSION: Transient AF complicating acute inferior MI is associated with an increased future risk of AF occurrence and ischemic stroke in patients with preserved LVEF, despite the use of antiplatelet therapy. PMID- 17400658 TI - Racial differences in cancer risk among relatives of patients with early onset lung cancer. AB - BACKGROUND: Relatives of patients with early onset lung cancer are at increased risk for lung cancer, and this risk varies by race. This study evaluates whether first-degree relatives of patients with early onset lung cancer are at increased risk for cancer at sites other than lung. METHODS: Family histories were ascertained from 673 lung cancer patients < 50 years of age identified from the Metropolitan Detroit Surveillance, Epidemiology and End Results program, and 773 age-, race-, and sex-matched control subjects were obtained via random-digit dialing. Data were collected for 3,556 case relatives (mothers, fathers, and siblings) and 3,943 control relatives, and unconditional logistic regression models using generalized estimating equations were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Among case relatives, African Americans were 2.44-fold more likely to have head and neck cancers and 1.86-fold more likely to have any tobacco-related cancer compared to white case relatives (95% CI, 1.04 to 5.69% and 95% CI, 1.25 to 2.76, respectively). African-American case relatives were at increased risk for head and neck cancers (OR, 13.42; 95% CI, 1.65 to 109.01), all tobacco-related cancers (OR, 3.77; 95% CI, 2.16 to 6.55), tobacco-related cancers other than lung (OR, 4.10; 95% CI, 1.56 to 10.79), and cancer at any site (OR, 1.45, 95% CI, 1.04 to 2.02) compared to African American control relatives. CONCLUSIONS: These results can be used to counsel family members of patients with early onset lung cancer, and suggest target populations for preventive strategies, including smoking cessation and appropriate screening. PMID- 17400659 TI - Chronic interstitial pneumonia in silicosis and mix-dust pneumoconiosis: its prevalence and comparison of CT findings with idiopathic pulmonary fibrosis. AB - BACKGROUND: Increased prevalence of chronic interstitial pneumonia (CIP) is reported in dust-exposed subjects. We investigated the prevalence of CIP in silicosis and mixed-dust pneumoconiosis and sought morphologic differences of CIP between the pneumoconiosis and idiopathic pulmonary fibrosis (IPF). METHODS: We reviewed CT scans of 243 silicosis and mixed-dust pneumoconiosis patients to identify any cases of parenchymal lung lesions showing a CIP pattern, and compared the CT findings with those of 62 patients with IPF. Two observers independently scored CT images and classified the CT pattern as typical or not typical for IPF. Differences were sought between the groups using a nonparametric test, Fisher exact test, and a logistic regression analysis. A radiopathologic correlation was performed in 11 pneumoconiosis patients. RESULTS: Twenty-eight patients (11.5%) showed CIP on CT. Seven patients (25%) showed a pattern not typical of IPF, while the remaining patients showed a pattern typical of IPF, 11 of which were confirmed pathologically. The extent of fibrosis did not differ between the groups; however, patients with pneumoconiosis showed less traction bronchiectasis (odds ratio [OR], 0.19; 95% confidence interval [CI], 0.08 to 0.48; p < 0.001), more subpleural homogeneous attenuation (OR, 2.56; 95% CI, 1.55 to 4.23; p < 0.001), and fibrosis was more randomly distributed (OR, 315.38; 95% CI, 4.68 to 21244.63; p = 0.007). Pathologically, subpleural homogeneous attenuation corresponded to dense fibrosis often with abundant silicotic nodules. CONCLUSIONS: Prevalence of CIP in pneumoconiosis was approximately 12% on CT. One fourth of patients showed an atypical IPF pattern, and the others showed a typical IPF pattern. PMID- 17400660 TI - The effects of helium-hyperoxia on 6-min walking distance in COPD: a randomized, controlled trial. AB - BACKGROUND: We hypothesized that breathing helium-hyperoxia (HeO2) would significantly improve 6-min walking test (6MWT) distance in COPD subjects. METHODS: This was a blinded, randomized crossover study. At visit 1, we assessed pulmonary function, exercise capacity, and 6MWT distance. Visits 2 and 3 consisted of four 6MWTs in which the following different inspired gases were used: room air (RA) by mask; 100% O2 by mask (mask O2); 100% O2 by nasal prongs (nasal O2); and 70% He/30% O2 by mask (HeO2). Walking distance, shortness of breath, leg fatigue, O2 saturation, and heart rate (HR) were assessed. RESULTS: Sixteen COPD subjects participated (mean FEV(1)/FVC ratio [+/- SD], 48 +/- 8%; mean FEV1, 55 +/- 13% predicted). Subjects walked farther when breathing HeO2 (564 m) compared to RA (497 m; p < 0.001), mask O2 (520 m; p < 0.001), or nasal O2 (528 m; p < 0.001). Despite the increased distance walked while breathing HeO2, there was no increase in shortness of breath or leg fatigue. There was desaturation when breathing RA (8%; p < 0.001) and nasal O2 (5%; p < 0.001), which was reduced when breathing HeO2 (3%; difference not significant) and mask O(2) (0%; difference not significant). There were no significant differences in HR in the four 6MWTs. CONCLUSIONS: The use of HeO2 increased 6MWT distance in COPD subjects more than either mask O2 or nasal O2 compared to RA. The increased walking distance was not associated with increased shortness of breath or leg fatigue. The results suggest that clinical benefit would be obtained by administering HeO2 during exercise, which may have significant clinical implications for the management of COPD patients. PMID- 17400661 TI - Treatment of chronic pulmonary aspergillosis by voriconazole in nonimmunocompromised patients. AB - BACKGROUND: There is no recognized medical treatment for chronic pulmonary aspergillosis (CPA) apart from surgery in patients with simple aspergilloma. To evaluate the efficacy of voriconazole in this setting, we conducted a retrospective multicenter study over a 3-year period. METHODS: For inclusion in the study, patients had to have received voriconazole for treatment of confirmed or probable CPA with a follow-up of at least 6 months. Clinical, radiologic, and mycologic data were collected at baseline, every 2 to 3 months, and at the end of treatment or at the date point. RESULTS: Twenty-four patients were included in the study, among which 9 patients presented with chronic cavitary pulmonary aspergillosis and 15 presented with chronic necrotizing pulmonary aspergillosis (CNPA). Voriconazole was given as a first-line treatment to 13 patients. The median duration of treatment and follow-up were 6.5 and 10 months, respectively. Three patients had to stop treatment with voriconazole because of toxicity. Symptoms and imagery findings were improved in 16 of 24 patients and 17 of 24 patients, respectively, at the end of follow-up. Mycology, which was positive at baseline in 21 of 23 patients, was negative in 18 of 19 patients at the end of follow-up; serologic test results were also negative in 6 of 19 evaluable patients, all of whom had CNPA. Improved radioclinical findings and mycologic eradication were observed at the end of follow-up in 11 of 19 patients (58%). Patients in whom the disease was controlled had a significantly longer median duration of treatment than patients in whom it was uncontrolled (9 vs 6 months, respectively; p = 0.04). CONCLUSION: Voriconazole provides effective treatment of CPA with an acceptable level of toxicity. PMID- 17400662 TI - Atmospheric pressure influences the risk of pneumothorax: beware of the storm! AB - BACKGROUND: Idiopathic spontaneous pneumothorax (ISP) results from rupture of blebs, bullas, or diseased alveolar walls. Initiating mechanisms may relate to increased transpulmonary pressure. The possible impact of changes in atmospheric pressure (Patm) on the occurrence of ISP remains uncertain. METHODS: We studied the relationship between the occurrence of ISP and meteorological conditions during a 4-year period in the urban area of Bologna, Italy, in which all cases of pneumothorax can be exhaustively identified. For each day of the study period, Patm and ambient temperature were obtained from the local meteorological institute. A cluster was defined as the admission of at least two patients with pneumothorax within 3 days of each other. RESULTS: There were 294 episodes of ISP; 247 (84%) occurred in 76 clusters. Clusters were significantly associated with wider differences in Patm between the index day (ie, the first day of the cluster) and the previous day (ie, the difference in mean [+/- SEM] Patm, -1.23 +/- 0.45 vs + 0.04 +/- 0.12 mm Hg, respectively; p = 0.01[analysis of variance]). Similarly, pneumothorax and storms (but not temperature) were significantly associated (p < 0.0001 [chi(2) test]). CONCLUSIONS: This large-scale study shows that patients with ISP are hospitalized in clusters and suggests that important variations in Patm may be involved. The knowledge of this relationship may help to understand the pathophysiology of the disease. PMID- 17400663 TI - Reproducibility of right ventricular volumes and ejection fraction using real time three-dimensional echocardiography: comparison with cardiac MRI. AB - OBJECTIVES: The nongeometric nature of the right ventricle (RV) makes it difficult to measure. We sought to determine whether real-time three-dimensional echocardiography (RT3DE) is superior to two-dimensional echocardiography (2DE) for the follow-up of RV function by validation vs cardiac MRI. METHODS: RV volumes and ejection fraction (EF) were studied with 2DE (including area-length [A-L], the modified two-dimensional subtraction [2DS] method, and the Simpson method of discs), RT3DE, and MRI in 50 patients with left ventricular wall motion abnormalities, the results of which suggested possible RV infarction. Test-retest variation was performed by a complete restudy using a separate sonographer within 24 h without the alteration of hemodynamics or therapy. Interobserver and intraobserver variations were noted in a subgroup of 20 patients. RESULTS: EF estimations were similar using each technique. The mean (+/- SD) MRI end diastolic volume (87 +/- 22 mL) was only slightly underestimated by RT3DE (mean difference, -3 +/- 10; p < 0.05), with a greater mean difference for 2DE A-L (-29 +/- 10; p < 0.05), and the Simpson method of discs (-29 +/- 23; p < 0.05), and was greatly overestimated by 2DS (mean difference, 26 +/- 23; p < 0.05). Similarly, the mean MRI end-systolic volume (46 +/- 17 mL) was only slightly underestimated by RT3DE (-4 +/- 7; p < 0.05), compared with 2DE A-L (-16 +/- 8; p < 0.05) and the Simpson method of discs (-16 +/- 8; p < 0.05), and was overestimated by 2DS (14 +/- 13; p < 0.05). RT3DE findings had a higher correlation with each parameter than any 2DE technique. There was also good intraobserver and interobserver correlation between RT3DE by two sonographers. RT3DE had less test-retest variation of RV volumes and EF than any 2DE measure. CONCLUSIONS: RT3DE is more accurate than two-dimensional approaches and reduces the test-retest variation of RV volumes and EF measurements in follow-up RV assessment. PMID- 17400664 TI - World Trade Center "sarcoid-like" granulomatous pulmonary disease in New York City Fire Department rescue workers. AB - BACKGROUND: Previous reports suggest that sarcoidosis occurs with abnormally high frequency in firefighters. We sought to determine whether exposure to World Trade Center (WTC) "dust" during the collapse and rescue/recovery effort increased the incidence of sarcoidosis or "sarcoid-like" granulomatous pulmonary disease (SLGPD). METHODS: During the 5 years after the WTC disaster, enrollees in the Fire Department of New York (FDNY) WTC monitoring and treatment programs who had chest radiograph findings suggestive of sarcoidosis underwent evaluation, including the following: chest CT imaging, pulmonary function, provocative challenge, and biopsy. Annual incidence rates were compared to the 15 years before the WTC disaster. RESULTS: After WTC dust exposure, pathologic evidence consistent with new-onset sarcoidosis was found in 26 patients: all 26 patients had intrathoracic adenopathy, and 6 patients (23%) had extrathoracic disease. Thirteen patients were identified during the first year after WTC dust exposure (incidence rate, 86/100,000), and 13 patients were identified during the next 4 years (average annual incidence rate, 22/100,000; as compared to 15/100,000 during the 15 years before the WTC disaster). Eighteen of 26 patients (69%) had findings consistent with asthma. Eight of 21 patients (38%) agreeing to challenge testing had airway hyperreactivity (AHR), findings not seen in FDNY sarcoidosis patients before the WTC disaster. CONCLUSION: After the WTC disaster, the incidence of sarcoidosis or SLGPD was increased among FDNY rescue workers. This new information about the early onset of WTC-SLGPD and its association with asthma/AHR has important public health consequences for disease prevention, early detection, and treatment following environmental/occupational exposures. PMID- 17400665 TI - Acute pulmonary embolism is an independent predictor of adverse events in severe decompensated heart failure patients. AB - BACKGROUND: Congestive heart failure (CHF) is a well-recognized risk factor for venous thromboembolism (VTE) and is associated with higher mortality in patients with an acute pulmonary embolism (PE). There are very few data on how acute PE affects the clinical course of patients with heart failure. The purpose of this study was to determine the impact of an acute PE on the short-term prognosis of patients hospitalized for decompensated CHF. METHODS: This was a prospective cohort study of 198 patients admitted to a coronary care unit between July 2001 and March 2003 with severe decompensated CHF. The primary outcome measure was death or rehospitalization at 3 months. RESULTS: PE was confirmed in 18 of 198 patients enrolled (9.1%). The groups with and without PE were comparable with regards to demographics, the prevalence of comorbid conditions, and severity of CHF (p > 0.05). The prevalence of cancer (p = 0.0001), previous VTE (p = 0.003), and right ventricular overload (p = 0.006) was higher in the PE group. The presence of PE was also associated with a longer hospital stay (37.5 +/- 71.6 days vs 15.4 +/- 15.0 days, p = 0.001) [mean +/- SD] and a higher incidence of death or rehospitalization at 3 months (72.2% vs 43.9%, p = 0.02). In a multiple logistic regression analysis, PE remained an independent predictor of death or rehospitalization at 3 months (odds ratio, 4.0; 95% confidence interval, 1.1 to 15.1; p = 0.038). CONCLUSIONS: Acute PE commonly complicates the hospital course of patients with severe CHF, increasing the length of hospital stay and the chance of death or rehospitalization at 3 months. PMID- 17400666 TI - The effect of neonatal bacille Calmette-Guerin vaccination on purified protein derivative skin test results in Canadian aboriginal children. AB - BACKGROUND: The effect that neonatal bacille Calmette-Guerin (BCG) vaccination has on tuberculin skin test (TST) results is not well evaluated in preschool children. METHODS: This was a retrospective cohort study of TST results in aboriginal children in Saskatchewan reserve communities. Records from the centralized provincial tuberculosis program were searched for aboriginal children aged 0 to 4 years during the time period 1991 to 1999. Only the first TST result reported as part of infant and preschool screening programs was considered. Children with active tuberculosis and those evaluated as part of a contact tracing program were excluded. The BCG-vaccinated and unvaccinated groups were compared using wheal size cut points of 5 mm, 10 mm, and 15 mm. RESULTS: Data from 1,086 children with neonatal BCG vaccination and 1,867 unvaccinated children were analyzed. The rate of TST reactions was higher in vaccinated children at all ages, using a cut point of 5 mm. The rate of TST reactions was no different in vaccinated children >or= 1 year old when using a cut point of 15 mm. When using a cut point of 10 mm, the rate of TST reactions was higher at age 1 year but not different at age 4 years in the vaccinated children. CONCLUSION: The rate of TST reactions in preschool aboriginal children living on a reserve who have received neonatal BCG vaccination is affected by the cut point and age. The BCG vaccination status and age should therefore be considered when interpreting TST reactivity in the clinical assessment of aboriginal children participating in a tuberculosis control program. PMID- 17400667 TI - Acute exacerbation of interstitial pneumonia other than idiopathic pulmonary fibrosis. AB - BACKGROUND: Acute exacerbation (AE) in idiopathic pulmonary fibrosis (IPF) is increasingly recognized as a relatively common and highly morbid clinical event. However, clinical data on AE in non-IPF interstitial pneumonia are sparse. This study was performed to find the frequency, clinical features, and outcome of AE in non-IPF interstitial pneumonia. METHODS: Retrospective analysis of 10 patients who satisfied the modified Akira criteria for AE during follow-up of 74 patients with surgical lung biopsy-confirmed idiopathic nonspecific interstitial pneumonia (I-NSIP) and 93 patients with biopsy-confirmed interstitial pneumonia associated with collagen vascular disease (CVD-IP). RESULTS: AE occurred in six patients with I-NSIP (1-year frequency, 4.2%) and in four patients with CVD-IP (rheumatoid arthritis [RA], n = 3; scleroderma, n = 1), with 1-year frequency of 3.3%. Median age was 58 years (range, 47 to 75); six patients were female. AE occurred in two patients immediately after surgical biopsy. Median duration of acute symptom before hospital admission was 10 days (range, 1 to 30). Median ratio of Pao(2) to the fraction of inspired oxygen (Fio(2)) was 172 (range, 107 to 273), and Pao(2)/Fio(2) ratio was < 200 in six patients. Surgical lung biopsy performed at the time of AE in two patients revealed diffuse alveolar damage superimposed on nonspecific interstitial pneumonia pattern. Four patients with I-NSIP survived to discharge and were followed up for 24 months (range, 6 to 121). CONCLUSION: AE occurred in the patients with I-NSIP with apparently better prognosis. In patients with CVD-IP, AE occurred mostly with RA-usual interstitial pneumonia in our small series with poor outcome. PMID- 17400668 TI - Misdiagnosis of community-acquired pneumonia and inappropriate utilization of antibiotics: side effects of the 4-h antibiotic administration rule. AB - BACKGROUND: The 2003 Infectious Diseases Society of America guidelines for community-acquired pneumonia (CAP) recommend the initiation of antibiotic therapy within 4 h of hospitalization. This quality indicator has been linked to the incentive compensation of third-party payers to hospitals. We evaluated the impact of this recommendation on the diagnosis of CAP and the utilization of antibiotics. METHODS: All patients with a hospital admission diagnosis of CAP before publication of the guidelines (January to June 2003) and after publication of the guidelines (January-June 2005) were included. We collected data on clinical signs and symptoms on presentation, chest radiograph findings, blood cultures prior to therapy with antibiotics, time to antibiotic administration, pneumonia severity index (PSI) score, confusion, urea, respiratory rate, BP, and age >or= 65 years (CURB-65), and mortality. RESULTS: A total of 518 patients were included in the study. More patients in 2005 had a hospital admission diagnosis of CAP without radiographic abnormalities compared to 2003 (2005, 91 patients [28.5%]; 2003, 41 patients [20.6%]; p = 0.04), and more patients received antibiotics within 4 h of triage (2005, 210 patients [65.8%]; 2003, 107 patients [53.8%]; p = 0.007). Blood cultures prior to antibiotic administration increased (2005, 220 patients [69.6%]; 2003, 93 patients [46.7%]; p < 0.001). However, the final diagnosis of CAP dropped to 58.9% in 2005 from 75.9% in 2003 (p < 0.001). The mean (+/- SD) antibiotic utilization per patient increased to 1.66 +/- 0.54 in 2005 compared to 1.39 +/- 0.58 in 2003 (p < 0.001). There were no significant differences in PSI or CURB-65 scores, or mortality. CONCLUSIONS: Linking antibiotic administration within 4 h of hospital admission (as a quality indicator) to financial compensation may result in an inaccurate diagnosis of CAP, inappropriate utilization of antibiotics, and thus less than optimal care. PMID- 17400669 TI - Fresh-frozen plasma and platelet transfusions are associated with development of acute lung injury in critically ill medical patients. AB - BACKGROUND: Transfusion has long been identified as a risk factor for acute lung injury (ALI)/ARDS. No study has formally evaluated the transfusion of specific blood products as a risk factor for ALI/ARDS in critically ill medical patients. METHOD: In this single-center retrospective cohort study, 841 consecutive critically ill patients were studied for the development of ALI/ARDS. Patients who received blood product transfusions were compared with those who did not, in univariate and multivariate propensity analyses. RESULTS: Two hundred ninety eight patients (35%) received blood transfusions. Transfused patients were older (mean [+/- SD] age, 67 +/- 17 years vs 62 +/- 19 years; p < 0.001) and had higher acute physiologic and chronic health evaluation (APACHE) III scores (74 +/- 32 vs 58 +/- 23; p < 0.001) than those who had not received transfusions. ALI/ARDS developed more commonly (25% vs 18%; p = 0.025) in patients exposed to transfusion. Seventeen patients received massive RBC transfusions (ie, > 10 U of blood transfused within 24 h), of whom 13 also received fresh-frozen plasma (FFP) and 11 received platelet transfusions. When adjusted for the probability of transfusion and other ALI/ARDS risk factors, any transfusion was associated with the development of ALI/ARDS (odds ratio [OR], 2.14; 95% confidence interval [CI], 1.24 to 3.75). Among those patients receiving individual blood products, ALI/ARDS was more likely to develop in patients who received FFP transfusions (OR, 2.48; 95% CI, 1.29 to 4.74) and platelet transfusions (OR, 3.89; 95% CI, 1.36 to 11.52) than in those who received only RBC transfusions (OR, 1.39; 95% CI, 0.79 to 2.43). CONCLUSION: Transfusion is associated with an increased risk of the development of ALI/ARDS in critically ill medical patients. The risk is higher with transfusions of plasma-rich blood products, FFP, and platelets, than with RBCs. PMID- 17400670 TI - Electromagnetic navigation diagnostic bronchoscopy in peripheral lung lesions. AB - BACKGROUND: Electromagnetic navigation bronchoscopy (ENB) with biopsy under fluoroscopic guidance has enhanced the yield of flexible bronchoscopy in the diagnosis of peripheral lung lesions. However, the accuracy of ENB navigation suggests that the addition of fluoroscopy is redundant. OBJECTIVES: Data were prospectively collected to determine the yield of ENB without fluoroscopy in the diagnosis of peripheral lung lesions. METHOD: ENB was performed via flexible bronchoscopy (superDimension/Bronchus system; superDimension Inc; Plymouth, MN). Biopsy specimens were obtained through the extended working channel after navigation. Fluoroscopy was not utilized, but post-transbronchial biopsy chest radiographs were obtained to exclude pneumothorax. The primary end point was diagnostic yield, and the secondary end points were navigation accuracy, procedure duration, and safety. Analysis by lobar distribution was also performed to assess performance in different lobes of the lung. RESULTS: Ninety-two peripheral lung lesions were biopsied in the 89 subjects. The diagnostic yield of ENB was 67%, which was independent of lesion size. Total procedure time ranged from 16.3 to 45.0 min (mean [+/- SD] procedure time, 26.9 +/- 6.5 min). The mean navigation error was 9 +/- 6 mm (range, 1 to 31 mm). There were two incidences of pneumothorax for which no intervention was required. When analyzed by lobar distribution, there was a trend toward a higher ENB yield in diagnosing lesions in the right middle lobe (88%). CONCLUSIONS: ENB can be used as a stand-alone bronchoscopic technique without compromising diagnostic yield or increasing the risk of pneumothorax. This may result in sizable timesaving and avoids radiation exposure. PMID- 17400671 TI - Registry and survival study in chinese patients with idiopathic and familial pulmonary arterial hypertension. AB - BACKGROUND: To evaluate the clinical features and survival data of patients with idiopathic pulmonary arterial hypertension (PAH) and familial PAH in Chinese patients. METHODS: Seventy-two patients with idiopathic PAH and familial PAH were enrolled in the study from 1999 to 2004 and were classified into two groups according to World Health Organization (WHO) functional class (I/II and III/IV). Clinical and hemodynamic data were recorded. RESULTS: The mean age of the 72 patients was 35.9 years with female patient/male patient ratio of 2.4:1. A significant difference was identified in the clinical presentation between two WHO functional class groups at baseline. Echocardiography showed a mean pulmonary systolic pressure of 98 mm Hg. Left ventricular end-diastolic diameter was significantly smaller in the group of patients in WHO functional class III/IV than in those in class I/II group. After follow-up for a mean (+/- SD) duration of 40.1 +/- 20.0 months, the survival rates at 1, 2, 3, and 5 years were 68.0%, 56.9%, 38.9%, and 20.8%, respectively. A significant difference was identified in survival rate between the class I/II and class III/IV groups (p = 0.02 [log rank test]). CONCLUSIONS: The baseline characteristics and survival rates of our cohort study are close to those of the National Institutes of Health Registry in the 1980s, and the 1-year survival rate is obviously lower for patients in this registry than for those in the French registry between 2002 to 2003. Lack of effective treatment was the main cause of poor survival in this study. Our results support the need of an appropriate treatment strategy for this devastating disease in China. PMID- 17400672 TI - Atrial septostomy decreases sympathetic overactivity in pulmonary arterial hypertension. AB - BACKGROUND: We have reported previously that the sympathetic nervous system is activated in patients with pulmonary arterial hypertension (PAH), and that this is only partly explained by a decrease in arterial oxygenation. Possible causes for increased muscle sympathetic nerve activity (MSNA) in patients with PAH include right atrial distension and decreased cardiac output. Both may be improved by atrial septostomy, but this intervention also further decreases arterial oxygenation. In the present study, we wanted to investigate the effect of atrial septostomy on MSNA in patients with PAH. METHODS: We recorded BP, heart rate (HR), arterial O2 saturation (SaO2), and MSNA before and after atrial septostomy in PAH patients (mean [+/- SE] age, 48 +/- 5 years) and in closely matched control subjects. Measurements were also performed after septostomy, while SaO2 was brought to the preprocedure level by supplemental O2 therapy. RESULTS: Compared to the control subjects (n = 10), the PAH patients (n = 11) had a lower mean BP (75 +/- 2 vs 96 +/- 3 mm Hg, respectively; p < 0.001), lower mean SaO2 (92 +/- 1% vs 97 +/- 0%, respectively; p < 0.001), increased mean HR (84 +/- 4 vs 68 +/- 3 beats/min; p < 0.01), and markedly increased mean MSNA (76 +/- 5 vs 29 +/- 2 bursts per minute; p < 0.001). Atrial septostomy decreased mean SaO2 (to 85 +/- 2%; p < 0.001) and mean MSNA (to 69 +/- 4 bursts per minute; p < 0.01), but did not affect HR or BP. Therapy with supplemental O2 did not affect MSNA, BP, or HR. The decrease in MSNA was correlated to the decrease in right atrial pressure (r = 0.62; p < 0.05). CONCLUSIONS: Atrial septostomy in PAH patients decreases sympathetic hyperactivity despite an associated decrease in arterial oxygenation, and this appears to be related to decreased right atrial distension. PMID- 17400673 TI - Significance of multiple carcinoid tumors and tumorlets in surgical lung specimens: analysis of 28 patients. AB - BACKGROUND: The clinical significance of multiple carcinoid tumorlets in surgical lung specimens has not been systematically analyzed. We reviewed our experience to determine the range of clinical circumstances associated with this finding. METHODS: We reviewed clinical records, available imaging, and pathology materials from patients evaluated at Mayo Clinic Rochester (from 1987 to 2000) with two or more carcinoid tumors or tumorlets in lung specimens. RESULTS: Twenty-eight of 294 patients with a diagnosis of carcinoid tumor or tumorlet had two or more lesions. Twenty-six patients (93%) were women; mean age was 65 years. Patients were categorized into three groups: multiple nodules (n = 17), solitary lung nodules on preoperative imaging (n = 7), and airflow limitation (n = 4). Approximately half of patients with multiple nodules had respiratory complaints; two patients had Cushing syndrome. Ten patients (58.8%) were suspected of having pulmonary metastases, including 7 patients with previously diagnosed malignancies. Intrathoracic lymph node metastases were present in three patients, none of whom had recurrent disease. One patient had a carcinoid tumor resected 8 years later. Extrathoracic metastases developed in another patient 3 years after presentation, and the patient was alive with disease 2 years later. Only one patient with airflow limitation had a syndrome resembling diffuse idiopathic pulmonary neuroendocrine cell hyperplasia. CONCLUSIONS: Our series represents the largest compilation of multiple carcinoid tumors or tumorlets. Our analysis reveals that multiple carcinoid tumors or tumorlets occur most commonly in patients with multiple nodules resembling metastatic disease. Significant airflow limitation is rare. Long-term survival is excellent, although patients have persistent disease. PMID- 17400674 TI - Internet-based self-management offers an opportunity to achieve better asthma control in adolescents. AB - BACKGROUND: Internet and short message service are emerging tools for chronic disease management in adolescents, but few data exist on the barriers to and benefits of internet-based asthma self-management. Our objective was to reveal the barriers and benefits perceived by adolescents with well-controlled and poorly controlled asthma to current and internet-based asthma management. METHODS: Ninety-seven adolescents with mild-to-moderate persistent asthma monitored their asthma control on a designated Web site. After 4 weeks, 35 adolescents participated in eight focus groups. Participants were stratified in terms of age, gender, and asthma control level. We used qualitative and quantitative methods to analyze the written focus group transcripts. RESULTS: Limited self-efficacy to control asthma was a significant barrier to current asthma management in adolescents with poor asthma control (65%) compared to adolescents with good asthma control (17%; p < 0.01). The former group revealed the following several benefits from internet-based asthma self-management: feasible electronic monitoring; easily accessible information; e-mail communication; and use of an electronic action plan. Personal benefits included the ability to react to change and to optimize asthma control. Patients with poor asthma control were able and ready to incorporate internet-based asthma self management for a long period of time (65%), whereas patients with good control were not (11%; p < 0.01). CONCLUSIONS: Our findings reveal a need for the support of self-management in adolescents with poorly controlled asthma that can be met by the application of novel information and communication technologies. Internet based self-management should therefore target adolescents with poor asthma control. PMID- 17400675 TI - The potential of a 2Tone Trainer to help patients use their metered-dose inhalers. AB - BACKGROUND: Many patients have problems using the correct inhalation technique when they use their metered-dose inhalers (MDIs). We have investigated whether a training aid (2Tone Trainer [2T]; Canday Medical Ltd; Newmarket, UK) helps to maintain the correct inhaler technique after patients leave the clinic METHODS: Ethics committee approval was obtained, and patients gave consent. Asthmatic patients who had been prescribed an MDI had their inhalation technique assessed. Their peak inhalation flow (PIF) when using their MDI, FEV(1), and the Juniper asthma quality of life questionnaire (AQLQ) score were measured. Those patients using the recommended MDI technique were the good-technique (GT) group. The remainder were randomized to receive verbal training (VT) or VT plus the 2T to improve their MDI technique. All patients returned 6 weeks later. RESULTS: There were 36, 35, and 36 asthmatic patients, respectively, who completed the GT, VT, and 2T procedures. FEV1 did not change within all groups between visit 1 and 2. PIF and AQLQ score did not change in the GT group. In the VT and 2T groups, the AQLQ score increased by mean differences of 0.33 (95% confidence interval [CI], 0.14 to 0.53; p < 0.001) and 0.74 (95% CI, 0.62 to 0.86; p < 0.001). At visit 1, all patients in the VT and 2T groups inhaled > 90 L/min decreasing to 12 patients and 1 patient, respectively, at visit 2 (p < 0.001 both groups). The overall changes in the 2T group for PIF and AQLQ score, between visits 1 and 2, were significantly (p < 0.001) greater than the corresponding changes in the VT group. CONCLUSION: The 2T helps patients to maintain the recommended MDI technique posttraining with improvements in AQLQ score. PMID- 17400676 TI - Panton-Valentine Leukocidin-positive methicillin-resistant Staphylococcus aureus lung infection in patients with cystic fibrosis. AB - BACKGROUND: Panton-Valentine Leukocidin-expressing (PVL+) methicillin-resistant Staphylococcus aureus (MRSA) is an emerging pathogen worldwide causing fatal necrotizing pneumonias in otherwise healthy individuals but has not been described in patients with cystic fibrosis (CF). Following two cases of patients with CF admitted with lung abscesses in association with PVL+ MRSA, we examined the incidence and the clinical characteristics of MRSA acquisition in our CF patient population. METHODS: Newly acquired MRSA isolates from patients with CF followed up at St. Louis Children's Hospital were analyzed for the presence of Panton-Valentine leukocidin coding region, clindamycin susceptibility, staphylococcal cassette chromosome (SCC) mec type, and multilocus sequence type. Medical records and pulmonary function studies at the time of MRSA isolation were reviewed. RESULTS: MRSA isolates from 40 CF patients were available for analysis. Six children (15%) had PVL+ MRSA infection. All PVL+ organisms were clindamycin susceptible. Patients who acquired a PVL+ organism were more likely to have a focal pulmonary infiltrate on chest radiograph, including cavitary lung lesions in two patients (p = 0.04), a markedly greater decline in FEV1 at the time of MRSA detection (p = 0.01), and a significantly higher WBC count (p = 0.04) and absolute neutrophil count (p = 0.04). These patients were more likely to be admitted for IV antibiotic therapy for respiratory illnesses (p < 0.01). CONCLUSIONS: We describe the emergence of PVL+ MRSA in our CF population in association with development of invasive lung infections including lung abscesses. Early identification and treatment of CF patients with newly acquired PVL+ MRSA may be crucial. PMID- 17400677 TI - Peak expiratory flow is not a quality indicator for spirometry: peak expiratory flow variability and FEV1 are poorly correlated in an elderly population. AB - BACKGROUND: Peak forced expiratory flow (PEF) and FEV(1) are spirometry measures used in diagnosing and monitoring lung diseases. We tested the premise that within-test variability in PEF is associated with corresponding variability in FEV(1) during a single test session. METHODS: A total of 2,464 healthy adults from the Health, Aging, and Body Composition Study whose spirometry results met American Thoracic Society acceptability criteria were screened and analyzed. The three "best" test results (highest sum of FVC and FEV(1)) were selected for each subject. For those with acceptable spirometry results, two groups were created: group 1, normal FEV(1)/FVC ratio; group 2, reduced FEV(1)/FVC ratio. For each subject, the difference between the highest and lowest PEF (DeltaPEF) and the associated difference between the highest and lowest FEV(1) (DeltaFEV(1)) were calculated. Regression analysis was performed using the largest PEF and best FEV(1), and the percentage of DeltaPEF (%DeltaPEF) and percentage of DeltaFEV(1) (%DeltaFEV(1)) were calculated in both groups. RESULTS: Regression analysis for group 1 and group 2 showed an insignificant association between %DeltaPEF and %DeltaFEV(1) (r(2) = 0.0001, p = 0.59, and r(2) = 0.040, p = 0.15, respectively). For both groups, a 29% DeltaPEF was associated with a 1% DeltaFEV(1). CONCLUSION: Within a single spirometry test session, %DeltaPEF and %DeltaFEV(1) contain independent information. PEF has a higher degree of intrinsic variability than FEV(1). Changes in PEF do not have a significant effect on FEV(1). Spirometry maneuvers should not be excluded based on peak flow variability. PMID- 17400678 TI - Airway nitric oxide in patients with cystic fibrosis is associated with pancreatic function, Pseudomonas infection, and polyunsaturated fatty acids. AB - BACKGROUND: Airway nitric oxide (NO) is low or normal in cystic fibrosis (CF) patients. This may affect bacterial status since NO has antimicrobial properties. Arachidonic acid (AA), which is increased in the serum and airways of CF patients, has been shown to reduce NO levels. The aim of this study was to investigate whether airway NO level correlates with genotype and pancreatic function, and whether low airway NO level is associated with bacterial infection and increased serum AA level in CF patients. METHOD: Nasal NO (nNO) and exhaled NO (eNO) were measured according to the European Respiratory Society/American Thoracic Society standard in 59 CF patients aged 7 to 55 years, 80% of whom were pancreatic insufficient (PI) and 51% were chronically infected with Pseudomonas aeruginosa. RESULTS: PI CF patients had significantly lower nNO levels than pancreatic-sufficient (PS) patients. Airway NO level did not correlate with lung function or inflammatory parameters. PI patients chronically infected with P aeruginosa had significantly lower nNO levels than noninfected PI patients. nNO level correlated inversely with the AA/docosahexaenoic acid ratio, and eNO with the essential fatty acid (FA) deficiency index, which is the ratio between mead acid and AA. CONCLUSIONS: CF patients with PI, which is associated with more severe genotypes, had lower airway NO levels than patients with PS. Low NO level was correlated to chronic P aeruginosa infection, and an association was found between airway NO level and the abnormal serum phospholipid FA pattern. PMID- 17400679 TI - Evaluation of persistent pulmonary hypertension after acute pulmonary embolism. AB - BACKGROUND: Better knowledge of the evolution of persistent pulmonary hypertension after acute pulmonary embolism (PE) is required to optimize the indication and timing of pulmonary endarterectomy (PEA). METHODS: We reviewed our experience with 17 consecutive patients demonstrated to have persistent pulmonary hypertension after acute massive (n = 1), submassive (n = 7), or recurrent PE (n = 9). RESULTS: After a median of 18 weeks of anticoagulation (range, 12 to 30 weeks) since the last PE, 10 patients showed residual pulmonary artery systolic pressure (PAsP) > 50 mm Hg. These patients demonstrated a significant progression in PAsP over the ensuing 6 to 12 months, from 73 +/- 14 to 101 +/- 26 mm Hg (p = 0.005) [mean +/- SD], and eight patients were found to be suitable candidates for PEA. In contrast, among seven patients with residual PAsP from 35 to 40 mm Hg (n = 3) and 41 to 50 mm Hg (n = 4), six patients had evidence of residual perfusion defects on the ventilation/perfusion scan and CT. The PAsP did not change significantly over the ensuing 6 to 12 months, except in two patients who had new episodes of acute PE. CONCLUSIONS: Two groups of patients can be identified based on the degree of residual pulmonary hypertension after acute PE. Patients with residual PAsP > 50 mm Hg should be evaluated for PEA since their pulmonary artery pressures will significantly progress over the ensuing 6 to 12 months despite the absence of recurrent PE. In contrast, patients with PAsP from 35 to 50 mm Hg are at risk for severe pulmonary hypertension if new PE occurs, and should therefore be closely monitored. PMID- 17400680 TI - Prevalence of gastroesophageal reflux disease in patients with nontuberculous mycobacterial lung disease. AB - BACKGROUND: Knowledge of the relationship between respiratory disorders and gastroesophageal reflux disease (GERD) is increasing. However, the association between GERD and pulmonary disease caused by nontuberculous mycobacteria (NTM) has not been studied in detail. We investigated the prevalence of GERD in patients with the nodular bronchiectatic form of NTM lung disease. METHODS: Fifty eight patients with the nodular bronchiectatic form of NTM lung disease underwent ambulatory 24-h esophageal pH monitoring. Of the 58 patients, 27 patients were identified as having Mycobacterium avium complex infection (15 with Mycobacterium intracellulare and 12 with M avium), and 31 patients had Mycobacterium abscessus pulmonary infection. RESULTS: The prevalence of GERD in patients with the nodular bronchiectatic form of NTM lung disease was 26% (15 of 58 patients). Only 27% (4 of 15 patients) had typical GERD symptoms. No statistically significant differences were found between patients with GERD and those without GERD with regard to age, sex, body mass index, or pulmonary function test results. However, patients with GERD were more likely to have a sputum smear that was positive for acid-fast bacilli (12 of 15 patients, 80%), compared with patients without GERD (19 of 43 patients, 44%) [p = 0.033]. In addition, bronchiectasis and bronchiolitis were observed in more lobes in patients with GERD than in patients without GERD (p = 0.008 and p = 0.005, respectively). CONCLUSIONS: Patients with the nodular bronchiectatic form of NTM lung disease have a high prevalence of increased esophageal acid exposure, usually without typical GERD symptoms. PMID- 17400681 TI - Beneficial effect of bilevel positive airway pressure on left ventricular function in ambulatory patients with idiopathic dilated cardiomyopathy and central sleep apnea-hypopnea: a preliminary study. AB - BACKGROUND: Sleep-disordered breathing is common in individuals with left ventricular (LV) dysfunction and has been treated with nocturnal positive airway pressure. We investigated whether treatment of central sleep apnea-hypopnea with bilevel positive airway pressure (BPAP) in ambulatory patients with idiopathic dilated cardiomyopathy (IDCM) might improve LV function. METHODS: Fifty-two consecutive patients with IDCM who underwent both cardiac catheterization and standard polysomnography were enrolled in the study; individuals with obstructive sleep apnea syndrome were excluded. Subjects with an apnea-hypopnea index (AHI) >or= 20 episodes per hour were randomized to receive medical therapy either alone (n = 11) or together with BPAP (n = 10). RESULTS: LV end-diastolic pressure, pulmonary capillary wedge pressure, and plasma concentration of brain natriuretic peptide were significantly greater, and LV ejection fraction (LVEF) was significantly lower in patients with an AHI >or= 20/h (n = 21, 40.4%) than in those with an AHI < 20/h (n = 31, 59.6%). LVEF (30.5 +/- 1.6% vs 50.8 +/- 3.5%, p < 0.001) [mean +/- SE] and plasma concentration of brain natriuretic peptide (162.8 +/- 44.5 pg/mL vs 32.7 +/- 17.6 pg/mL, p = 0.02) were significantly increased and decreased, respectively, after treatment with BPAP (daily use, 4.8 +/- 0.3 h) for 3 months, whereas these parameters remained unchanged in the control subjects. CONCLUSIONS: Our findings suggest that treatment of coexisting central sleep apnea-hypopnea with BPAP improves LV function in ambulatory patients with IDCM. BPAP should thus be considered as a nonpharmacologic adjunct to conventional drug therapy in such patients. PMID- 17400682 TI - High prevalence of proximal and distal gastroesophageal reflux disease in advanced COPD. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) is common in a variety of chronic respiratory diseases, but little is known about GERD in the setting of COPD. The aims of this study were to determine the prevalence, presentation, and predictors of GERD based on proximal and distal esophageal pH monitoring in patients with severe COPD. METHODS: Forty-one COPD patients with a mean FEV1 of 24% of predicted underwent dual-probe 24-h esophageal pH monitoring, and 1 patient underwent esophagogastroduodenoscopy. RESULTS: The prevalence of GERD was 57%. Elevated distal and proximal reflux were present in 41% and 46% of patients undergoing esophageal pH studies, respectively. Fifteen percent of these patients had abnormal proximal reflux despite having normal distal probe results. Most patients with GERD were not receiving acid blockers at the time of their referral, and only one third reported heartburn and/or acid regurgitation during the pH study. Only higher body mass index was predictive of reflux on regression analysis (odds ratio, 1.2; 95% confidence interval, 1.0 to 1.5; p = 0.05). CONCLUSIONS: GERD is common in advanced COPD. Patients are often asymptomatic and have a relatively high prevalence of isolated abnormal proximal reflux. Dual probe monitoring is therefore well suited for detecting GERD in patients with advanced COPD. PMID- 17400683 TI - Diagnostic tests for asthma in firefighters. AB - BACKGROUND: Subjects with asthma do not meet medical requirements for professions such as firefighting. OBJECTIVE: To prospectively determine the diagnostic value of respiratory symptoms and various tests used in the assessment of asthma in a cohort of firefighters. METHODS: A questionnaire, spirometry, direct and indirect airway challenge tests, exhaled nitric oxide, and skin-prick tests were administered prospectively to 101 of 107 firefighters employed in Basel, Switzerland. Asthma was defined as the combination of respiratory symptoms with airway hyperresponsiveness. RESULTS: Six of 101 firefighters (6%) had physician diagnosed asthma, which could be confirmed in 4 firefighters. In contrast, asthma was diagnosed in 14% (14 of 101 firefighters). Wheezing was the most sensitive symptom for the diagnosis of asthma (sensitivity, 78%; specificity, 93%). Other respiratory symptoms showed a higher specificity than wheezing but a markedly lower sensitivity. Bronchial airway challenge with mannitol was the most sensitive (92%) and specific (97%) diagnostic test for asthma. Using a cutoff point of 47 parts per billion, nitric oxide had a similar specificity (96%) but lower sensitivity (42%) compared to the direct (methacholine) and indirect (mannitol) airway challenge tests. CONCLUSION: Asthma was considerably underdiagnosed in firefighters. The combination of a structured symptom questionnaire with a bronchial challenge test allows to identify patients with asthma and should routinely be used in the assessment of active firefighters and may be of help when evaluating candidates for this profession. PMID- 17400684 TI - Transition from IV epoprostenol to subcutaneous treprostinil in pulmonary arterial hypertension: a controlled trial. AB - BACKGROUND: We determined the relative efficacy of subcutaneous (SC) treprostinil in stable World Health Organization class II and III patients transitioned from IV epoprostenol. METHODS: This was an 8-week, multicenter, randomized study in which patients were transitioned from IV epoprostenol to SC treprostinil or placebo over a period of up to 14 days and monitored carefully during and after the transition period for signs of deterioration. Patients with clinical deterioration were returned promptly to epoprostenol. Placebo or SC treprostinil doses were titrated in response to symptoms. Time to adjudicated clinical deterioration was compared between treatment groups, and exercise capacity, symptoms of disease, and safety were assessed throughout the study. RESULTS: Twenty-two patients were enrolled and completed the study. Seven of 8 patients (88%) [corrected] withdrawn to placebo had clinical deterioration, while only 1 of 14 patients (7%) [corrected] withdrawn to SC treprostinil had clinical deterioration (p = 0.00023 based on a treatment comparison of time to deterioration). Analyses of exercise capacity and symptoms strongly supported the efficacy of SC treprostinil in epoprostenol-treated patients. Adverse events consisted of painful infusion site reactions and anticipated prostacyclin side effects. CONCLUSIONS: SC treprostinil is effective in pulmonary arterial hypertension and prevents clinical deterioration and maintains functional status in patients transitioned from epoprostenol. PMID- 17400685 TI - The incidence of recognized heparin-induced thrombocytopenia in a large, tertiary care teaching hospital. AB - BACKGROUND: Heparin-induced thrombocytopenia (HIT) is estimated to occur in up to 5% of patients receiving unfractionated heparin. The goal was to determine the incidence of HIT within our 1,061-bed tertiary care institution. METHODS: A retrospective review of three hospital database systems (ie, admission, pharmacy, and laboratory) was undertaken for a 1-year period ending in March 2004. The pharmacy database was queried to identify patients who received heparin and those who received a direct thrombin inhibitor (DTI). The medical records of patients receiving a DTI were reviewed to categorize the indication for DTI therapy. The laboratory system database was queried to retrieve heparin platelet factor 4 immunoassay results. RESULTS: A total of 58,814 patient admissions occurred with an estimated 24,068 patients being exposed to unfractionated heparin. DTI therapy was administered to 133 patients. Of these, 49 new HIT cases and 15 cases of suspected HIT (unconfirmed) were identified. The overall incidence of recognized new HIT was 0.2%. New HIT occurred in 0.76% of patients receiving therapeutic dose IV heparin and in < 0.1% of patients receiving antithrombotic prophylaxis (subcutaneous heparin). Forty-nine percent of all new HIT cases were in coronary artery bypass and/or valve replacement surgery patients, while no cases were identified in hip/knee arthroplasty patients. CONCLUSIONS: The incidence of recognized HIT in a large teaching institution was 0.2%, with a 0.76% incidence in those patients receiving therapeutic-dose IV heparin. The low incidence likely reflects a brief duration of heparin exposure for many patients. Approximately half of all new HIT cases were recognized in the cardiovascular surgery population. PMID- 17400686 TI - Sources of long-term variability in measurements of lung function: implications for interpretation and clinical trial design. AB - BACKGROUND: The objective of the study was to characterize the biological and technical components of variability associated with longitudinal measurements of FEV(1) and carbon monoxide diffusing capacity (Dlco). Variability was apportioned to subject and instrument for five commercially available pulmonary function testing (PFT) systems: Collins CPL (Ferraris Respiratory; Louisville, CO); Morgan Transflow Test PFT System (Morgan Scientific; Haverhill, MA); SensorMedics Vmax 22D (VIASYS Healthcare; Yorba Linda, CA); Jaeger USA Masterscreen Diffusion TP (VIASYS Healthcare; Yorba Linda, CA); and Medical Graphics Profiler DX System (Medical Graphics Corporation; St. Paul, MN). METHODS: This was a randomized, replicated cross-over, single-center methodology study in 11 healthy subjects aged 20 to 65 years. Spirometry and Dlco measurements were performed at baseline, 3 months, and 6 months. Repetitive simulations of FEV(1) and Dlco were performed on the same instruments on four occasions over a 90-day period using a spirometry waveform generator and a Dlco simulator. RESULTS: The coefficient of variation associated with repetitive measurements of FEV(1) or Dlco in subjects was consistently larger than that associated with repetitive simulated waveforms across the five instruments. Instrumentation accounted for 13 to 58% of the total FEV(1) and 36 to 70% of the total Dlco variability observed in subjects. Sample size estimates of hypothetical studies designed to detect treatment group differences of 0.050 L in FEV(1) and 0.5 mL/min/mm Hg in Dlco varied as much as four times depending on the instrument utilized. CONCLUSIONS: These results provide a semiquantitative assessment of the biological and technical components of PFT variability in a highly standardized setting. They illustrate how instrument choice and test variability can impact sample size determinations in clinical studies that use FEV(1) and Dlco as end points. PMID- 17400687 TI - Asthma in American Indian adults: the Strong Heart Study. AB - BACKGROUND: Despite growing recognition that asthma is an important cause of morbidity among American Indians, there has been no systematic study of this disease in older adults who are likely to be at high risk of complications related to asthma. Characterization of the impact of asthma among American Indian adults is necessary in order to design appropriate clinical and preventive measures. METHODS: A sample of participants in the third examination of the Strong Heart Study, a multicenter, population-based, prospective study of cardiovascular disease in American Indians, completed a standardized respiratory questionnaire, performed spirometry, and underwent allergen skin testing. Participants were > or = 50 years old. RESULTS: Of 3,197 participants in the third examination, 6.3% had physician-diagnosed asthma and 4.3% had probable asthma. Women had a higher prevalence of physician-diagnosed asthma than men (8.2% vs 3.2%). Of the 435 participants reported in the asthma substudy, morbidity related to asthma was high: among those with physician-diagnosed asthma: 97% reported trouble breathing and 52% had severe persistent disease. The mean FEV(1) in those with physician-diagnosed asthma was 61.3% of predicted, and 67.2% reported a history of emergency department visits and/or hospitalizations in the last year, yet only 3% were receiving regular inhaled corticosteroids. CONCLUSIONS: The prevalence of asthma among older American Indians residing in three separate geographic areas of the United States was similar to rates in other ethnic groups. Asthma was associated with low lung function, significant morbidity and health-care utilization, yet medications for pulmonary disease were underutilized by this population. PMID- 17400688 TI - Sleep and well-being of ICU housestaff. AB - BACKGROUND: Our understanding of the effect of the Accreditation Council for Graduate Medical Education (ACGME)-mandated work-hour limitation on physicians' quality of life, sleepiness, and sleep-work habits is evolving. In this study, we sought to determine the effect of work-hour reduction on quality of life in residents and fellows (ICU housestaff) when subject to the ACGME-compliant schedule of one institution. To determine the effect of work-hour reduction on subjective and objective measures of sleepiness in ICU housestaff at a center. METHODS: A single-center study of 34 residents and 10 fellows who were studied before and after the ACGME-mandated work-hour limitation went into effect in July 2003. RESULTS: In a single center, after the work-hour reduction, residents reported statistically significant but minor improvements in sleep time, subjective sleepiness, and some aspects of quality of life (p < 0.05). Both before and after work-hour limitations, subjective sleepiness and quality-of-life indexes deteriorated during the course of the ICU rotation. Following work-hour reductions, subjective sleepiness improved (p < 0.05), but objective sleepiness was unchanged (p = 0.6). Moreover, after the implementation of work-hour reductions, 59%, 43%, and 25% of the ICU team had mean sleep latency < 10, 7, and 5 min, respectively, with 14% of the team manifesting sleep-onset rapid eye movement periods (signifying severe sleepiness) before beginning their extended work-hour period. CONCLUSIONS: In ICU housestaff, at a single center, small benefits to quality of life and subjective sleepiness were realized by an ACGME compliant work-hour schedule. Significant levels of objective sleepiness, however, remained. Further measures may need to be undertaken to address the persistence of sleepiness in ICU housestaff. These findings may not be generalized outside of the scheduling system studied. PMID- 17400689 TI - Common variable immunodeficiency: association between memory B cells and lung diseases. AB - BACKGROUND: Malabsorption syndrome often develops in patients with common variable immunodeficiency (CVID). Why structural damages appear in some CVID patients and not in others is not fully understood. Memory B cells (MBs) are responsible for the production of specific antibodies, and their defects have previously been related to autoimmune, granulomatous, and lymphoproliferative complications of CVID. The objective of this study was to ascertain whether a relationship exists between MB defects and the clinical outcome of respiratory and intestinal involvement in these patients. METHODS: Forty-one CVID patients were grouped as follows, according to the quantification of peripheral MBs: the MB2 group (n = 7) included patients with normal MBs; the MB1 group (n = 16) included patients with low switched MBs; and the MB0 group (n = 18) included patients with absent/low MBs. The clinical outcome of respiratory and intestinal involvement of patients was then compared among the three groups. RESULTS: In the MB0 group, chronic lung disease (ie, bronchiectasis and diminished FVC and/or FEV1) developed in 50% of patients vs 13% in the MB1 group and 0% in the MB2 group (p < 0.05). In the MB0 group, malabsorption syndrome or chronic noninfectious diarrhea developed in 50% of patients vs 19% in the MB1 group and 0% in the MB2 group (p < 0.05). No differences were found among the three groups for age at onset of symptoms, delay in diagnosis/treatment, months of follow up/treatment, and prediagnostic serum IgG concentration. CONCLUSIONS: Alterations in MB count appear to be associated with a severe clinical outcome of respiratory and intestinal involvement in CVID. The MB count could be a useful laboratory parameter for orienting the prognosis and management of CVID patients. PMID- 17400690 TI - Pulmonary impairment after tuberculosis. AB - BACKGROUND: Pulmonary impairment subsequent to a cure of pulmonary tuberculosis has been described only in selected populations. METHODS: We compared pulmonary function in a case-control study of 107 prospectively identified patients with pulmonary tuberculosis who had completed at least 20 weeks of therapy and 210 patients with latent tuberculosis infection (LTBI). RESULTS: Both groups had similar risk factors for pulmonary impairment. Impairment was present in 59% of tuberculosis subjects and 20% of LTBI control subjects. FVC, FEV1, FEV1/FVC ratio, and the midexpiratory phase of forced expiratory flow were significantly lower in the treated pulmonary tuberculosis patients than in the comparison group. Ten patients with a history of pulmonary tuberculosis (9.4%) had less than half of their expected vital capacity vs one patient (0.53%) in the LTBI group. Another 42 patients (39%) with tuberculosis had between 20% and 50% of the expected vital capacity vs 36 patients with LTBI (17%). After adjusting for risk, survivors of tuberculosis were 5.4 times more likely to have abnormal pulmonary function test results than were LTBI patients (p > 0.001; 95% confidence interval, 2.98 to 9.68). Birth in the United States (odds ratio [OR], 2.64; p = 0.003) and age (OR, 1.03; p = 0.005) increased the odds of impairment. Pulmonary impairment was more common in cigarette smokers; however, after adjusting for demographic and other risk factors, the difference did not reach statistical significance (p = 0.074). CONCLUSIONS: These findings indicate that pulmonary impairment after tuberculosis is associated with disability worldwide and support more aggressive case prevention strategies and posttreatment evaluation. For many persons with tuberculosis, a microbiological cure is the beginning not the end of their illness. PMID- 17400691 TI - Atomic force microscope studies of the fusion of floating lipid bilayers. AB - This study investigated the fusion of apposing floating bilayers of egg L-alpha phosphatidylcholine (egg PC) or 1,2-dimyristoyl-sn-glycero-3-phosphocholine. Atomic force microscope measurements of fusion forces under different compression rates were acquired to reveal the energy landscape of the fusion process under varied lipid composition and temperature. Between compression rates of approximately 1000 and approximately 100,000 pN/s, applied forces in the range from approximately 100 to approximately 500 pN resulted in fusion of floating bilayers. Our atomic force microscope measurements indicated that one main energy barrier dominated the fusion process. The acquired dynamic force spectra were fit with a simple model based on the transition state theory with the assumption that the fusion activation potential is linear. A significant shift in the energy landscape was observed when bilayer fluidity and composition were modified, respectively, by temperature and different cholesterol concentrations (15% < or = chol < or = 25%). Such modifications resulted in a more than twofold increase in the width of the fusion energy barrier for egg PC and 1,2-dimyristoyl-sn-glycero 3-phosphocholine floating bilayers. The addition of 25% cholesterol to egg PC bilayers increased the activation energy by approximately 1.0 k(B)T compared with that of bilayers with egg PC alone. These results reveal that widening of the energy barrier and consequently reduction in its slope facilitated membrane fusion. PMID- 17400692 TI - Elastic properties of the cell surface and trafficking of single AMPA receptors in living hippocampal neurons. AB - Although various approaches are routinely used to study receptor trafficking, a technology that allows for visualizing trafficking of single receptors at the surface of living cells remains lacking. Here we used atomic force microscope to simultaneously probe the topography of living cells, record the elastic properties of their surface, and examine the distribution of transfected alpha amino-3-hydroxy-5-methylisoxazole-4-proprionic acid (AMPA)-type glutamate receptors (AMPAR). On nonstimulated neurons, AMPARs were located in stiff nanodomains with high elasticity modulus relative to the remaining cell surface. Receptor stimulation with N-methyl-D-aspartate (NMDA) provoked a permanent disappearance of these stiff nanodomains followed by a decrease (53%) of the number of surface AMPARs. Blocking electrical activity before NMDA stimulation recruited the same number of AMPARs for internalization, preceded by the loss of the stiff nanodomains. However, in that case, the stiff nanodomains were recovered and AMPARs were reinserted into the membrane shortly after. Our results show that modulation of receptor distribution is accompanied by changes in the local elastic properties of cell membrane. We postulate, therefore, that the mechanical environment of a receptor might be critical to determine its specific distribution behavior in response to different stimuli. PMID- 17400693 TI - Pore formation in a lipid bilayer under a tension ramp: modeling the distribution of rupture tensions. AB - The rupture of fluid membrane vesicles with a steady ramp of micropipette suction has been shown to produce a distribution of breakage tensions, with a mean that rises rapidly with tension rate. Starting from a lattice model that incorporates the essential features of the lipid bilayers held together with hydrophobic forces, and developing it to handle varying tension rates, we reproduce the main features of the experimental results. In essence, we show that the rupture kinetics are driven by the nucleation and growth of pores, with two limiting kinetics-growth-limited and nucleation-limited. The model has been extended to address the role of peptides in solution that can adsorb and insert themselves into the bilayer. At concentrations below those required to spontaneously rupture the membrane, the effect of the peptides is to lower the rupture tensions systematically for all tension rates. PMID- 17400694 TI - A steady-state model of spreading depression predicts the importance of an unknown conductance in specific dendritic domains. AB - Spreading depression (SD) is a pathological wave of transient neuronal inactivation. We recently reported that the characteristic sustained complete depolarization is restricted to specific cell domains where the input resistance (R(in)) first becomes negligible before achieving partial recovery, whereas in adjacent, more polarized membranes it drops by much less. The experimental study of the participating membrane channels is hindered by their mixed contribution and heterogeneous distribution. Therefore, we derived a biophysical model to analyze the conductances that replicate the subcellular profile of R(in) during SD. Systematic variation of conductance densities far beyond the ranges reported failed to fit the experimental values. Besides standard potassium, sodium, and Glu-mediated conductances, the initial opening and gradual closing of an as yet undetermined large conductance is required to account for the evolution of R(in). Potassium conductances follow in the relative contribution and their closing during the late phase is also predicted. Large intracellular potential gradients from zero to rest are readily sustained between shunted and adjacent SD-spared membranes, which remain electroregenerative. The gradients are achieved by a combination of high-conductance subcellular domains and transmembrane ion redistribution in extended but discrete dendritic domains. We conclude that the heterogeneous subcellular behavior is due to local membrane properties, some of which may be specifically activated under extreme SD conditions. PMID- 17400695 TI - Intrinsic fluctuations, robustness, and tunability in signaling cycles. AB - Covalent modification cycles (e.g., phosphorylation-dephosphorylation) underlie most cellular signaling and control processes. Low molecular copy number, arising from compartmental segregation and slow diffusion between compartments, potentially renders these cycles vulnerable to intrinsic chemical fluctuations. How can a cell operate reliably in the presence of this inherent stochasticity? How do changes in extrinsic parameters lead to variability of response? Can cells exploit these parameters to tune cycles to different ranges of stimuli? We study the dynamics of an isolated phosphorylation cycle. Our model shows that the cycle transmits information reliably if it is tuned to an optimal parameter range, despite intrinsic fluctuations and even for small input signal amplitudes. At the same time, the cycle is sensitive to changes in the concentration and activity of kinases and phosphatases. This sensitivity can lead to significant cell-to-cell response variability. It also provides a mechanism to tune the cycle to transmit signals in various amplitude ranges. Our results show that signaling cycles possess a surprising combination of robustness and tunability. This combination makes them ubiquitous in eukaryotic signaling, optimizing signaling in the presence of fluctuations using their inherent flexibility. On the other hand, cycles tuned to suppress intrinsic fluctuations can be vulnerable to changes in the number and activity of kinases and phosphatases. Such trade-offs in robustness to intrinsic and extrinsic fluctuations can influence the evolution of signaling cascades, making them the weakest links in cellular circuits. PMID- 17400696 TI - Reparameterization of all-atom dipalmitoylphosphatidylcholine lipid parameters enables simulation of fluid bilayers at zero tension. AB - Molecular dynamics simulations of dipalmitoylphosphatidylcholine (DPPC) lipid bilayers using the CHARMM27 force field in the tensionless isothermal-isobaric (NPT) ensemble give highly ordered, gel-like bilayers with an area per lipid of approximately 48 A(2). To obtain fluid (L(alpha)) phase properties of DPPC bilayers represented by the CHARMM energy function in this ensemble, we reparameterized the atomic partial charges in the lipid headgroup and upper parts of the acyl chains. The new charges were determined from the electron structure using both the Mulliken method and the restricted electrostatic potential fitting method. We tested the derived charges in molecular dynamics simulations of a fully hydrated DPPC bilayer. Only the simulation with the new restricted electrostatic potential charges shows significant improvements compared with simulations using the original CHARMM27 force field resulting in an area per lipid of 60.4 +/- 0.1 A(2). Compared to the 48 A(2), the new value of 60.4 A(2) is in fair agreement with the experimental value of 64 A(2). In addition, the simulated order parameter profile and electron density profile are in satisfactory agreement with experimental data. Thus, the biologically more interesting fluid phase of DPPC bilayers can now be simulated in all-atom simulations in the NPT ensemble by employing our modified CHARMM27 force field. PMID- 17400697 TI - Coarse-grained strategy for modeling protein stability in concentrated solutions. III: directional protein interactions. AB - We extend our coarse-grained modeling strategy described in parts I and II of this investigation to account for nonuniform spatial distributions of hydrophobic residues on the solvent-exposed surfaces of native proteins. Within this framework, we explore how patchy surfaces can influence the solvent-mediated protein-protein interactions, and the unfolding and self-assembly behaviors of proteins in solution. In particular, we compare the equilibrium unfolding and self-assembly trends for three model proteins that share the same overall sequence hydrophobicity, but exhibit folded configurations with different solvent exposed native-state surface morphologies. Our model provides new insights into how directional interactions can affect native-state protein stability in solution. We find that strongly-directional attractions between native molecules with patchy surfaces can help stabilize the folded conformation through the formation of self-assembled clusters. In contrast, native proteins with more uniform surfaces are destabilized by protein-protein attractions involving the denatured state. Finally, we discuss how the simulation results provide insights into the experimental solution behaviors of several proteins that display directional interactions in their native states. PMID- 17400698 TI - The effects of force inhibition by sodium vanadate on cross-bridge binding, force redevelopment, and Ca2+ activation in cardiac muscle. AB - Strongly bound, force-generating myosin cross-bridges play an important role as allosteric activators of cardiac thin filaments. Sodium vanadate (Vi) is a phosphate analog that inhibits force by preventing cross-bridge transition into force-producing states. This study characterizes the mechanical state of cross bridges with bound Vi as a tool to examine the contribution of cross-bridges to cardiac contractile activation. The K(i) of force inhibition by Vi was approximately 40 microM. Sinusoidal stiffness was inhibited with Vi, although to a lesser extent than force. We used chord stiffness measurements to monitor Vi induced changes in cross-bridge attachment/detachment kinetics at saturating [Ca(2+)]. Vi decreased chord stiffness at the fastest rates of stretch, whereas at slow rates chord stiffness actually increased. This suggests a shift in cross bridge population toward low force states with very slow attachment/detachment kinetics. Low angle x-ray diffraction measurements indicate that with Vi cross bridge mass shifted away from thin filaments, implying decreased cross bridge/thin filament interaction. The combined x-ray and mechanical data suggest at least two cross-bridge populations with Vi; one characteristic of normal cycling cross-bridges, and a population of weak-binding cross-bridges with bound Vi and slow attachment/detachment kinetics. The Ca(2+) sensitivity of force (pCa(50)) and force redevelopment kinetics (k(TR)) were measured to study the effects of Vi on contractile activation. When maximal force was inhibited by 40% with Vi pCa(50) decreased, but greater force inhibition at higher [Vi] did not further alter pCa(50). In contrast, the Ca(2+) sensitivity of k(TR) was unaffected by Vi. Interestingly, when force was inhibited by Vi k(TR) increased at submaximal levels of Ca(2+)-activated force. Additionally, k(TR) is faster at saturating Ca(2+) at [Vi] that inhibit force by > approximately 70%. The effects of Vi on k(TR) imply that k(TR) is determined not only by the intrinsic properties of the cross-bridge cycle, but also by cross-bridge contribution to thin filament activation. PMID- 17400699 TI - Low-resolution structure and fluorescence anisotropy analysis of protein tyrosine phosphatase eta catalytic domain. AB - The rat protein tyrosine phosphatase eta, rPTPeta, is a class I "classical" transmembrane RPTP, with an intracellular portion composed of a unique catalytic region. The rPTPeta and the human homolog DEP-1 are downregulated in rat and human neoplastic cells, respectively. However, the malignant phenotype is reverted after exogenous reconstitution of rPTPeta, suggesting that its function restoration could be an important tool for gene therapy of human cancers. Using small-angle x-ray scattering (SAXS) and biophysical techniques, we characterized the intracellular catalytic domain of rat protein tyrosine phosphatase eta (rPTPetaCD) in solution. The protein forms dimers in solution as confirmed by SAXS data analysis. The SAXS data also indicated that rPTPetaCD dimers are elongated and have an average radius of gyration of 2.65 nm and a D(max) of 8.5 nm. To further study the rPTPetaCD conformation in solution, we built rPTPetaCD homology models using as scaffolds the crystallographic structures of RPTPalpha D1 and RPTPmicro-D1 dimers. These models were, then, superimposed onto ab initio low-resolution SAXS structures. The structural comparisons and sequence alignment analysis of the putative dimerization interfaces provide support to the notion that the rPTPetaCD dimer architecture is more closely related to the crystal structure of autoinhibitory RPTPalpha-D1 dimer than to the dimeric arrangement exemplified by RPTPmicro-D1. Finally, the characterization of rPTPetaCD by fluorescence anisotropy measurements demonstrates that the dimer dissociation is concentration dependent with a dissociation constant of 21.6 +/- 2.0 microM. PMID- 17400700 TI - Coarse-grained peptide modeling using a systematic multiscale approach. AB - A systematic new approach to derive multiscale coarse-grained (MS-CG) models has been recently developed. The approach employs information from atomistically detailed simulations to derive CG forces and associated effective potentials. In this work, the MS-CG methodology is extended to study two peptides representing distinct structural motifs, alpha-helical polyalanine and the beta-hairpin V(5)PGV(5). These studies represent the first known application of this approach to peptide systems. Good agreement between the MS-CG and atomistic models is achieved for several structural properties including radial distribution functions, root mean-square deviation, and radius of gyration. The new MS-CG models are able to preserve the native states of these peptides within approximately 1 A backbone root mean-square deviation during CG simulations. The MS-CG approach, as with most coarse-grained models, has the potential to increase the length and timescales accessible to molecular simulations. However, it is also able to maintain a clear connection to the underlying atomistic-scale interactions. PMID- 17400701 TI - Detection of protein orientation on the silica microsphere surface using transverse electric/transverse magnetic whispering gallery modes. AB - The state of adsorbed protein molecules can be examined by comparing the shifts in a narrow line resonance wavelength of transverse electric (TE) and transverse magnetic (TM) whispering gallery modes (WGM) when the molecules adsorb onto a transparent microsphere that houses WGM. In adsorption of bovine serum albumin (BSA) onto an aminopropyl-modified silica microsphere, the TM/TE shift ratio indicated highly anisotropic polarizability of BSA in the direction normal to the surface, most likely ascribed to anchoring the heart-shaped protein molecule by one of its tips. The polarization-dependent resonance shift was confirmed when the surrounding refractive index was uniformly changed by adding salt, which would simulate adsorption of large objects. PMID- 17400702 TI - A new approach to the detection and statistical classification of Ca2+ sparks. AB - The availability of high-speed, two-dimensional (2-D) confocal microscopes and the expanding armamentarium of fluorescent probes presents unprecedented opportunities and new challenges for studying the spatial and temporal dynamics of cellular processes. The need to remove subjectivity from the detection process, the difficulty of the human eye to detect subtle changes in fluorescence in these 2-D images, and the large volume of data produced by these confocal microscopes call for the need to develop algorithms to automatically mark the changes in fluorescence. These fluorescence signal changes are often subtle, so the statistical estimate of the likelihood that the detected signal is not noise is an integral part of the detection algorithm. This statistical estimation is fundamental to our new approach to detection; in earlier Ca(2+) spark detectors, this statistical assessment was incidental to detection. Importantly, the use of the statistical properties of the signal local to the spark, instead of over the whole image, reduces the false positive and false negative rates. We developed an automatic spark detection algorithm based on these principles and used it to detect sparks on an inhomogeneous background of transverse tubule-labeled rat ventricular cells. Because of the large region of the cell surveyed by the confocal microscope, we can detect a large enough number of sparks to measure the dynamic changes in spark frequency in individual cells. We also found, in contrast to earlier results, that cardiac sparks are spatially symmetric. This new approach puts the detection of fluorescent signals on a firm statistical foundation. PMID- 17400703 TI - Diffusion of insulin-like growth factor-I and ribonuclease through fibrin gels. AB - A fluorescence-based method for simultaneously determining the diffusion coefficients of two proteins is described, and the diffusion coefficient of insulin-like growth factor (IGF-I) and ribonuclease (RNase) in a 0.27% fibrin hydrogel is reported. The method is based on two-color imaging of the relaxation of the protein concentration field with time and comparing the results with a transport model. The gel is confined in a thin (200 microm) capillary and the protein is labeled with a fluorescent dye. The experimentally determined diffusion coefficient of RNase (D = 1.21 x 10(-6) cm(2)/s) agrees with literature values for dilute gels and bulk aqueous solutions, thus indicating the gel and the dye had a negligible effect on diffusion. The experimental diffusion coefficient of IGF-I (D = 1.59 x 10(-6) cm(2)/s), in the absence of binding to the fibrin matrix, is consistent with the dimensions of the molecule known from x ray crystallography and a correlation between D and molecular weight based on 14 other proteins. The experimental method developed here holds promise for determining molecular transport properties of biomolecules under a variety of conditions, for example, when the molecule adsorbs to the gel or is convected through the gel by fluid transport. PMID- 17400704 TI - Tracking single Kinesin molecules in the cytoplasm of mammalian cells. AB - Understanding dynamic cellular processes requires precise knowledge of the distribution, transport, and interactions of individual molecules in living cells. Despite recent progress in in vivo imaging, it has not been possible to express and directly track single molecules in the cytoplasm of live cells. Here, we overcome these limitations by combining fluorescent protein-labeling with high resolution total internal reflection fluorescence microcopy, using the molecular motor Kinesin-1 as model system. First, we engineered a three-tandem monomeric Citrine tag for genetic labeling of individual molecules and expressed this motor in COS cells. Detailed analysis of the quantized photobleaching behavior of individual fluorescent spots demonstrates that we are indeed detecting single proteins in the cytoplasm of live cells. Tracking the movement of individual cytoplasmic molecules reveals that individual Kinesin-1 motors in vivo move with an average speed of 0.78 +/- 0.11 microm/s and display an average run length of 1.17 +/- 0.38 microm, which agrees well with in vitro measurements. Thus, Kinesin 1's speed and processivity are not upregulated or hindered by macromolecular crowding. Second, we demonstrate that standard deviation maps of the fluorescence intensity computed from single molecule image sequences can be used to reveal important physiological information about infrequent cellular events in the noisy fluorescence background of live cells. Finally, we show that tandem fluorescent protein tags enable single-molecule, in vitro analyses of extracted, mammalian expressed proteins. Thus, by combining direct genetic labeling and single molecule imaging in vivo, our work establishes an important new biophysical method for observing single molecules expressed and localized in the mammalian cytoplasm. PMID- 17400705 TI - Modeling of the role of a Bax-activation switch in the mitochondrial apoptosis decision. AB - We performed in silico modeling of the regulatory network of mitochondrial apoptosis through which we examined the role of a Bax-activation switch in governing the mitochondrial apoptosis decision. Two distinct modeling methods were used in this article. One is continuous and deterministic, comprised of a set of ordinary differential equations. The other, carried out in a discrete manner, is based on a cellular automaton, which takes stochastic fluctuations into consideration. We focused on dynamic properties of the mitochondrial apoptosis regulatory network. The roles of Bcl-2 family proteins in cellular responses to apoptotic stimuli were examined. In our simulations, a self amplification process of Bax-activation is indicated. Further analysis suggests that the core module of Bax-activation is bistable in both deterministic and stochastic models, and this feature is robust to noise and wide ranges of parameter variation. When coupling with Bax-polymerization, it forms a one-way switch, which governs irreversible behaviors of Bax-activation even with attenuation of apoptotic stimulus. Together with the growing biochemical evidence, we propose a novel molecular switch mechanism embedded in the mitochondrial apoptosis regulatory network and give a plausible explanation for the all-or-none, irreversible character of mitochondrial apoptosis. PMID- 17400706 TI - Impairment of the photorespiratory pathway accelerates photoinhibition of photosystem II by suppression of repair but not acceleration of damage processes in Arabidopsis. AB - Oxygenation of ribulose-1,5-bisphosphate catalyzed by Rubisco produces glycolate 2-P. The photorespiratory pathway, which consists of photorespiratory carbon and nitrogen cycles, metabolizes glycolate-2-P to the Calvin cycle intermediate glycerate-3-P and is proposed to be important for avoiding photoinhibition of photosystem II (PSII), especially in C3 plants. We show here that mutants of Arabidopsis (Arabidopsis thaliana) with impairment of ferredoxin-dependent glutamate synthase, serine hydroxymethyltransferase, glutamate/malate transporter, and glycerate kinase had accelerated photoinhibition of PSII by suppression of the repair of photodamaged PSII and not acceleration of the photodamage to PSII. We found that suppression of the repair process was attributable to inhibition of the synthesis of the D1 protein at the level of translation. Our results suggest that the photorespiratory pathway helps avoid inhibition of the synthesis of the D1 protein, which is important for the repair of photodamaged PSII upon interruption of the Calvin cycle. PMID- 17400707 TI - Light and metabolic signals control the selective degradation of sucrose synthase in maize leaves during deetiolation. AB - The content and activity of Suc (Suc) synthase (SUS) protein is high in sink organs but low in source organs. In this report, we examined light and metabolic signals regulating SUS protein degradation in maize (Zea mays) leaves during deetiolation. We found that SUS protein accumulated in etiolated leaves of the dark-grown seedlings but was rapidly degraded upon exposure to white, blue, or red light. This occurred concurrent with the accumulation of photosynthetic enzymes, such as Rubisco and Rubisco activase, and enzymes of Suc biosynthesis such as Suc-phosphate synthase. Deetiolation-induced SUS degradation was not inhibited by the proteasome inhibitor MG132. Moreover, neither full-length nor truncated SUS phosphorylated at the serine-170 site was found in the crude 26S proteasome fraction (150,000g postmicrosomal pellet) isolated in the presence of MG132. However, SUS degradation was strongly inhibited by feeding cycloheximide or amino acids to detached leaves, while Suc feeding had no effect. Of the amino acids tested, exogenous glutamate had the greatest effect. Collectively, these results demonstrate that SUS protein degradation during deetiolation: (1) is selective; (2) can be triggered by either blue- or red light-mediated signaling pathways; (3) does not involve the 26S proteasome; and (4) is inhibited by free amino acids. These findings suggest that SUS degradation is important to supply residues for the synthesis of other proteins required for autotrophic metabolism. PMID- 17400709 TI - Building bridges: integrative solutions for managing complex comorbid conditions. PMID- 17400710 TI - Elastin-insufficient mice show normal cardiovascular remodeling in 2K1C hypertension despite higher baseline pressure and unique cardiovascular architecture. AB - Mice heterozygous for the elastin gene (ELN(+/-)) show unique cardiovascular properties, including increased blood pressure and smaller, thinner arteries with an increased number of lamellar units. Some of these properties are also observed in humans with supravalvular aortic stenosis, a disease caused by functional heterozygosity of the elastin gene. The arterial geometry in ELN(+/-) mice is contrary to the increased thickness that would be expected in an animal demonstrating hypertensive remodeling. To determine whether this is due to a decreased capability for cardiovascular remodeling or to a novel adaptation of the ELN(+/-) cardiovascular system, we increased blood pressure in adult ELN(+/+) and ELN(+/-) mice using the two-kidney, one-clip Goldblatt model of hypertension. Successfully clipped mice have a systolic pressure increase of at least 15 mmHg over sham-operated animals. ELN(+/+) and ELN(+/-)-clipped mice show significant increases over sham-operated mice in cardiac weight, arterial thickness, and arterial cross-sectional area with no changes in lamellar number. There are no significant differences in most mechanical properties with clipping in either genotype. These results indicate that ELN(+/+) and ELN(+/-) hearts and arteries remodel similarly in response to adult induced hypertension. Therefore, the cardiovascular properties of ELN(+/-) mice are likely due to developmental remodeling in response to altered hemodynamics and reduced elastin levels. PMID- 17400712 TI - HMG Co-A reductase inhibitors improve microvascular outcomes in the metabolic syndrome independent of plasma total cholesterol. PMID- 17400711 TI - Transient middle cerebral artery occlusion causes different structural, mechanical, and myogenic alterations in normotensive and hypertensive rats. AB - Transient focal cerebral ischemia in the rat alters vessel properties, and spontaneously hypertensive rats (SHR) show a poorer outcome after ischemia. In the present study we examined the role of hypertension on vessel properties after ischemia-reperfusion. The right middle cerebral artery (MCA) was occluded (90 min) and reperfused (24 h) in SHR (n = 12) and Wistar-Kyoto rats (WKY; n = 11). Sham-operated rats (SHR, n = 10; WKY, n = 10) were used as controls. The structural, mechanical, and myogenic properties of the MCA were assessed by pressure myography. Nuclei distribution and elastin content and organization were analyzed by confocal microscopy. Infarct volume was larger in SHR than in WKY rats. Ischemia-reperfusion induced adventitial hypertrophy associated with an increase in the total number of adventitial cells. In addition, fenestrae area and arterial distensibility increased and myogenic tone decreased in the MCA of WKY rats after ischemia-reperfusion. Hypertension per se induced hypertrophic inward remodeling. Ischemia-reperfusion decreased the cross-sectional area of the MCA in SHR, without significant changes in distensibility, despite an increase in fenestrae area. In addition, MCA myogenic properties were not altered after ischemia-reperfusion in SHR. Our results indicate that in normotensive rats, MCA develops a compensatory mechanism (i.e., enhanced distensibility and decreased myogenic tone) that counteracts the effect of ischemia-reperfusion and ensures correct cerebral irrigation. These compensatory mechanisms are lost in hypertension, thereby explaining, at least in part, the greater infarct volume observed in SHR. PMID- 17400713 TI - Vascular endothelial growth factor-C stimulates the lymphatic pump by a VEGF receptor-3-dependent mechanism. AB - Vascular endothelial growth factor (VEGF)-C plays an important role in lymphangiogenesis; however, functional responses of lymphatic vessels to VEGF-C have not been characterized. We tested the hypothesis that VEGF-C-induced activation of VEGF receptor (VEGFR)-3 increases lymphatic pump output. We examined the in vivo pump activity of rat mesenteric collecting lymphatics using intravital microscopy during basal conditions and during treatment with 1 nM recombinant VEGF-C, the selective VEGFR-3 agonist VEGF-Cys(156)Ser mutation (C156S; 1 nM), or 0.1 nM VEGF-A. Their specific responses were also analyzed during selective inhibition of VEGFR-3 with MAZ-51. Contraction frequency, end diastolic diameter, end-systolic diameter, stroke volume index, pump flow index, and ejection fraction were evaluated. We also assessed arteriolar diameter and microvascular extravasation of FITC-albumin. The results show that both VEGF-C and VEGF-C156S significantly increased contraction frequency, end-diastolic diameter, stroke volume index, and pump flow index in a time-dependent manner. VEGF-A caused a different response characterized by a significantly increased stroke volume after 30 min of treatment. MAZ-51 (5 muM) caused tonic constriction and decreased contraction frequency. In addition, 0.5 and 5 muM MAZ-51 attenuated VEGF-C- and VEGF-C156S-induced lymphatic pump activation. VEGF-A caused vasodilation of arterioles, whereas VEGF-C and VEGF-C156S did not significantly alter arteriolar diameter. Also, VEGF-A and VEGF-C caused increased microvascular permeability, whereas VEGF-C156S did not. Our results demonstrate that VEGF-C increases lymphatic pumping through VEGFR-3. Furthermore, changes in microvascular hemodynamics are not required for VEGFR-3-mediated changes in lymphatic pump activity. PMID- 17400708 TI - Transcript profiling of poplar leaves upon infection with compatible and incompatible strains of the foliar rust Melampsora larici-populina. AB - To understand key processes governing defense mechanisms in poplar (Populus spp.) upon infection with the rust fungus Melampsora larici-populina, we used combined histological and molecular techniques to describe the infection of Populus trichocarpa x Populus deltoides 'Beaupre' leaves by compatible and incompatible fungal strains. Striking differences in host-tissue infection were observed after 48-h postinoculation (hpi) between compatible and incompatible interactions. No reactive oxygen species production could be detected at infection sites, while a strong accumulation of monolignols occurred in the incompatible interaction after 48 hpi, indicating a late plant response once the fungus already penetrated host cells to form haustorial infection structures. P. trichocarpa whole-genome expression oligoarrays and sequencing of cDNAs were used to determine changes in gene expression in both interactions at 48 hpi. Temporal expression profiling of infection-regulated transcripts was further compared by cDNA arrays and reverse transcription-quantitative polymerase chain reaction. Among 1,730 significantly differentially expressed transcripts in the incompatible interaction, 150 showed an increase in concentration > or =3-fold, whereas 62 were decreased by > or =3 fold. Regulated transcripts corresponded to known genes targeted by R genes in plant pathosystems, such as inositol-3-P synthase, glutathione S-transferases, and pathogenesis-related proteins. However, the transcript showing the highest rust-induced up-regulation encodes a putative secreted protein with no known function. In contrast, only a few transcripts showed an altered expression in the compatible interaction, suggesting a delay in defense response between incompatible and compatible interactions in poplar. This comprehensive analysis of early molecular responses of poplar to M. larici-populina infection identified key genes that likely contain the fungus proliferation in planta. Sequences from the SSH library described in this article can be retrieved in GenBank under accession numbers CT 027996 to CT 029994 and CT 033829. PMID- 17400714 TI - Cardioprotective effects of stretch are mediated by activation of sarcolemmal, not mitochondrial, ATP-sensitive potassium channels. AB - To determine whether sarcolemmal and/or mitochondrial ATP-sensitive potassium (K(ATP)) channels (sarcK(ATP), mitoK(ATP)) are involved in stretch-induced protection, isolated isovolumic rat hearts were assigned to the following protocols: nonstretched hearts were subjected to 20 min of global ischemia (Is) and 30 min of reperfusion, and before Is stretched hearts received 5 min of stretch + 10 min of no intervention. Stretch was induced by a transient increase in left ventricular end-diastolic pressure (LVEDP) from 10 to 40 mmHg. Other hearts received 5-hydroxydecanoate (5-HD; 100 microM), a selective inhibitor of mitoK(ATP), or HMR-1098 (20 microM), a selective inhibitor of sarcK(ATP), before the stretch protocol. Systolic function was assessed through left ventricular developed pressure (LVDP) and maximal rise in velocity of left ventricular pressure (+dP/dt(max)) and diastolic function through maximal decrease in velocity of left ventricular pressure (-dP/dt(max)) and LVEDP. Lactate dehydrogenase (LDH) release and ATP content were also measured. Stretch resulted in a significant increase of postischemic recovery and attenuation of diastolic stiffness. At 30 min of reperfusion LVDP and +dP/dt(max) were 87 +/- 4% and 92 +/ 6% and -dP/dt(max) and LVEDP were 95 +/- 9% and 10 +/- 4 mmHg vs. 57 +/- 6%, 53 +/- 6%, 57 +/- 10%, and 28 +/- 5 mmHg, respectively, in nonstretched hearts. Stretch increased ATP content and did not produce LDH release. 5-HD did not modify and HMR-1098 prevented the protection achieved by stretch. Our results show that the beneficial effects of stretch on postischemic myocardial dysfunction, cellular damage, and energetic state involve the participation of sarcK(ATP) but not mitoK(ATP). PMID- 17400715 TI - An analysis of the effects of stretch on IGF-I secretion from rat ventricular fibroblasts. AB - Mechanical force can induce a number of fundamental short- and long-term responses in myocardium. These include alterations in ECM, activation of cell signaling pathways, altered gene regulation, changes in cell proliferation and growth, and secretion of a number of peptides and growth factors. It is now known that a number of these autocrine/paracrine factors are secreted from both cardiomyocytes and ventricular cardiac fibroblasts (CFb) in response to stretch. One such substance is IGF-I. IGF-I is an important autocrine/paracrine factor that can regulate physiological or pathophysiological responses, such as hypertrophy. In this study, we addressed the possible effects of mechanical perturbation, biaxial strain, on IGF-I secretion from adult rat CFb. CFb were subjected to either static stretch (3-10%) or cyclic stretch (10%; 0.1-1 Hz) over a 24-h period. IGF-1 secretion from CFb in response to selected stretch paradigms was examined using ELISA to measure IGF-I concentrations in conditioned media. Static stretch did not result in any measurable modulation of IGF-I secretion from CFb. However, cyclic stretch significantly increased IGF-I secretion from CFb in a frequency- and time-dependent manner compared with nonstretched controls. This stretch-induced increase in secretion was relatively insensitive to changes in extracellular [Ca(2+)] or to block of L-type Ca(2+) channels. In contrast, thapsigargin, an inhibitor of sarco(endo)plasmic reticulum Ca(2+) ATPase, remarkably decreased stretch-induced IGF-I secretion from CFb. We further show that IGF-I can upregulate mRNA expression of atrial natriuretic peptide in myocytes. In summary, cyclic stretch can significantly increase IGF-I secretion from CFb, and this effect is dependent on a thapsigargin-sensitive pool of intracellular [Ca(2+)]. PMID- 17400716 TI - Alterations to myofibrillar protein function in nonischemic regions of the heart early after myocardial infarction. AB - Remote-zone left ventricular dysfunction (LVD) contributes to global reductions in contractile function after localized myocardial infarction (MI). However, the molecular mechanisms underlying this form of LVD are not clear. This study tested the hypothesis that myofibrillar protein function is directly affected in remote zone LVD early after MI. Cardiac myosin and native thin filaments were purified from mouse myocardium taken from both the nonnecrotic zone adjacent to and the nonischemic zone remote from an infarct induced by 1 h of coronary occlusion followed by 24 h of reperfusion. Thin filament velocities were measured using the in vitro motility assay. Results showed that overall function was significantly reduced in samples from both the adjacent (43 +/- 12% of control, n = 7) and remote (53 +/- 8% of control, n = 13) zones when compared with control proteins (P < 0.05). Myosin from the remote zone propelled control thin filaments at reduced velocities similar to those measured above. In contrast, the Ca(2+) sensitivity of remote-zone thin filaments over control myosin was unchanged from control thin filaments (half-maximal at pCa 6.32 +/- 0.08 and 6.27 +/- 0.06, respectively) but showed a 20% increase in velocity at saturating Ca(2+) that parallels an increase in tropomyosin phosphorylation. Myosin dysfunction may be related to oxidation of cysteines in the myosin heavy chains or carbonylation of myosin binding protein-C. We hypothesize that phosphorylation of tropomyosin may serve a compensatory role, augmenting contraction during periods of oxidative stress when myosin function is compromised. PMID- 17400717 TI - On the role of junctin in cardiac Ca2+ handling, contractility, and heart failure. AB - Junctin is a transmembrane protein located at the cardiac junctional sarcoplasmic reticulum (SR) and forms a quaternary complex with the Ca(2+) release channel, triadin and calsequestrin. Impaired protein interactions within this complex may alter the Ca(2+) sensitivity of the Ca(2+) release channel and may lead to cardiac dysfunction, including hypertrophy, depressed contractility, and abnormal Ca(2+) transients. To study the expression of junctin and, for comparison, triadin, in heart failure, we measured the levels of these proteins in SR from normal and failing human hearts. Junctin was below our level of detection in SR membranes from failing human hearts, and triadin was downregulated by 22%. To better understand the role of junctin in the regulation of Ca(2+) homeostasis and contraction of cardiac myocytes, we used an adenoviral approach to overexpress junctin in isolated rat cardiac myocytes. A recombinant adenovirus encoding the green fluorescent protein served as a control. Infection of myocytes with the junctin-expressing virus resulted in an increased RNA and protein expression of junctin. Ca(2+) transients showed a decreased maximum Ca(2+) amplitude, and contractility of myocytes was depressed. Our results demonstrate that an increased expression of junctin is associated with an impaired Ca(2+) homeostasis. Downregulation of junctin in human heart failure may thus be a compensatory mechanism. PMID- 17400718 TI - Reduced systolic wave generation and increased peripheral wave reflection in chronic heart failure. AB - In human heart failure the role of wave generation by the ventricle and wave reflection by the vasculature is contentious. The aim of this study was to compare wave generation and reflection in normal subjects with patients with stable compensated heart failure. Twenty-nine normal subjects and 67 patients with heart failure (New York Heart Association class II or III) were studied by noninvasive techniques applied to the common carotid artery. Data were analyzed by wave intensity analysis to determine the nature and direction of waves during the cardiac cycle. The energy carried by an early systolic forward compression wave (S wave) generated by the left ventricle and responsible for acceleration of flow in systole was significantly reduced in subjects with heart failure (P < 0.001), and the timing of the peak of this wave was delayed. In contrast, reflection of this wave was increased in subjects with heart failure (P < 0.001), but the timing of reflections with respect to the S wave was unchanged. The energy of an expansion wave generated by the heart in protodiastole was unaffected by heart failure. The carotid artery wave speed and the augmentation index did not significantly differ between subjects with heart failure compared with normal individuals. The ability of the left ventricle to generate a forward compression wave is markedly impaired in heart failure. Increased wave reflection serves to maintain systolic blood pressure but also places an additional load on cardiac function in heart failure. PMID- 17400719 TI - Cardiac effects of endothelin receptor antagonism in endotoxemic pigs. AB - Myocardial depression in sepsis is frequently encountered clinically and contributes to morbidity and mortality. Increased plasma levels of endothelin-1 (ET-1) have been described in septic shock, and previous reports have shown beneficial effects on cardiovascular performance and survival in septic models using ET receptor antagonists. The aim of the current study was to investigate specific cardiac effects of ET receptor antagonism in endotoxicosis. Sixteen domestic pigs were anesthetized and subjected to endotoxin for 5 h. Eight of these pigs were given tezosentan (dual ET receptor antagonist) after 3 h. Cardiac effects were evaluated using the left ventricular (LV) pressure-volume relationship. Endotoxin was not associated with any effects on parameters of LV contractile function [end-systolic elastance (Ees), preload recruitable stroke work (PRSW), power(max)/end-diastolic volume (PWR(max)/EDV) and dP/dt(max)/end diastolic volume (dP/dt(max)/EDV)] but with impairments in isovolumic relaxation (time constant for pressure decay, tau) and mechanical efficiency. Tezosentan administration decreased Ees, PWR(max)/EDV, and dP/dt(max)/EDV, while improving tau and LV stiffness. Thus, dual ET receptor antagonism was associated with a decline in contractile function but, in contrast, improved diastolic function. Positive hemodynamic effects from ET receptor antagonism in acute endotoxemia may be due to changes in cardiac load and enhanced diastolic function rather than improved contractile function. PMID- 17400720 TI - Impairment of cardiac insulin signaling and myocardial contractile performance in high-cholesterol/fructose-fed rats. AB - Although insulin resistance is recognized as a potent and prevalent risk factor for coronary heart disease, less is known as to whether insulin resistance causes an altered cardiac phenotype independent of coronary atherosclerosis. In this study, we investigated the relationship between insulin resistance and cardiac contractile dysfunctions by generating a new insulin resistance animal model with rats on high cholesterol-fructose diet. Male Sprague-Dawley rats were given high cholesterol-fructose (HCF) diet for 15 wk; the rats developed insulin resistance syndrome characterized by elevated blood pressure, hyperlipidemia, hyperinsulinemia, impaired glucose tolerance, and insulin resistance. The results show that HCF induced insulin resistance not only in metabolic-response tissues (i.e., liver and muscle) but also in the heart as well. Insulin-stimulated cardiac glucose uptake was significantly reduced after 15 wk of HCF feeding, and cardiac insulin resistance was associated with blunted Akt-mediated insulin signaling along with glucose transporter GLUT4 translocation. Basal fatty acid transporter FATP1 levels were increased in HCF rat hearts. The cardiac performance of the HCF rats exhibited a marked reduction in cardiac output, ejection fraction, stroke volume, and end-diastolic volume. It also showed decreases in left ventricular end-systolic elasticity, whereas the effective arterial elasticity was increased. In addition, the relaxation time constant of left ventricular pressure was prolonged in the HCF group. Overall, these results indicate that insulin resistance reduction of cardiac glucose uptake is associated with defects in insulin signaling. The cardiac metabolic alterations that impair contractile functions may lead to the development of cardiomyopathy. PMID- 17400721 TI - Inhibition of histone deacetylase on ventricular remodeling in infarcted rats. AB - Histone deacetylase (HDAC) determines the acetylation status of histones and, thereby, controls the regulation of gene expression. HDAC inhibitors have been shown to inhibit cardiomyocyte growth in vitro and in vivo. We assessed whether HDAC inhibitors exert a beneficial effect on the remodeling heart in infarcted rats. At 24 h after ligation of the left anterior descending artery, male Wistar rats were randomized to vehicle, HDAC inhibitors [valproic acid (VPA) and tributyrin], an agonist of HDAC (theophylline), VPA + theophylline, or tributyrin + theophylline for 4 wk. Significant ventricular hypertrophy was detected as increased myocyte size at the border zone isolated by enzymatic dissociation after infarction. Cardiomyocyte hypertrophy and collagen formation at the remote region and border zone were significantly attenuated by VPA and tributyrin with a similar potency compared with that induced by the vehicle. Left ventricular shortening fraction was significantly higher in the VPA- and tributyrin-treated groups than in the vehicle-treated group. Increased synthesis of atrial natriuretic peptide mRNA after infarction was confirmed by RT-PCR, consistent with the results of immunohistochemistry and Western blot for acetyl histone H4. The beneficial effects of VPA and tributyrin were abolished by theophylline, implicating HDAC as the relevant target. Inhibition of HDAC by VPA or tributyrin can attenuate ventricular remodeling after infarction. This might provide a worthwhile therapeutic target. PMID- 17400722 TI - Possible roles of neuropeptide Y Y3-receptor subtype in rat aortic endothelial cell proliferation under hypoxia, and its specific signal transduction. AB - The present study was undertaken to determine whether neuropeptide Y (NPY) induces proliferation of rat aortic endothelial cells (RAECs). Since NPY increased the permeability of RAEC monolayers to large molecules via the NPY Y(3) receptor, RAEC proliferation has been evaluated in terms of NPY-receptor subtypes and also intracellular mechanisms. RAECs were incubated with gases containing 20, 15, or 10% O(2) and a certain amount of N(2), depending on the O(2) content in 5% CO(2) incubators. NPY (10(-9)-10(-6) M) increased the RAEC numbers under hypoxic conditions, such as 15 or 10% O(2). Peptide YY elicited no proliferative effect on RAEC, and NPY-(18-36) inhibited the NPY-induced increase in cell number, suggesting that NPY increases the RAEC count through the NPY Y(3) receptor. Pertussis toxin, U-73122, GF-109203X, myristorylated autocamtide-2-related inhibitory peptide, and wortmannin inhibited the NPY-induced proliferation of RAEC concentration dependently. DY9760e little affected the proliferation caused by NPY. ML-9 and imatinib actually enhanced the NPY-induced proliferation of cells. These results indicated that the NPY Y(3) receptor is coupled with G(i) protein, and that NPY-induced increases in RAEC proliferation are mediated by phospholipase C-protein kinase C and/or phosphatidylinositol 3-kinase pathways. In intracellular Ca(2+)-calmodulin-dependent pathways, calmodulin-dependent protein kinase II partly participates in the NPY-induced cell proliferation. Regarding the previously reported effect of NPY on the permeability of RAEC monolayers to large molecules, it is probable that protein kinase C and phosphatidylinositol 3-kinase pathways are activated for both permeability and cell proliferation induced by NPY under hypoxia, relevant to new insights into the roles of NPY in ischemia-hypoxia. PMID- 17400723 TI - Role of genetic and environmental influences on heart rate variability in middle aged men. AB - Our aim was to estimate causal relationships of genetic factors and different specific environmental factors in determination of the level of cardiac autonomic modulation, i.e., heart rate variability (HRV), in healthy male twins and male twins with chronic diseases. The subjects were 208 monozygotic (MZ, 104 healthy) and 296 dizygotic (DZ, 173 healthy) male twins. A structured interview was used to obtain data on lifetime exposures of occupational loading, regularly performed leisure-time sport activities, coffee consumption, smoking history, and chronic diseases from 12 yr of age through the present. A 5-min ECG at supine rest was recorded for the HRV analyses. In univariate statistical analyses based on genetic models with additive genetic, dominance genetic, and unique environmental effects, genetic effects accounted for 31-57% of HRV variance. In multivariate statistical analysis, body mass index, percent body fat, coffee consumption, smoking, medication, and chronic diseases were associated with different HRV variables, accounting for 1-11% of their variance. Occupational physical loading and leisure-time sport activities did not account for variation in any HRV variable. However, in the subgroup analysis of healthy and diseased twins, occupational loading explained 4% of the variability in heart periods. Otherwise, the interaction between health status and genetic effects was significant for only two HRV variables. In conclusion, genetic factors accounted for a major portion of the interindividual differences in HRV, with no remarkable effect of health status. No single behavioral determinant appeared to have a major influence on HRV. The effects of medication and diseases may mask the minimal effect of occupational loading on HRV. PMID- 17400724 TI - Effect of intracellular Ca2+ and action potential duration on L-type Ca2+ channel inactivation and recovery from inactivation in rabbit cardiac myocytes. AB - Ca(2+) current (I(Ca)) recovery from inactivation is necessary for normal cardiac excitation-contraction coupling. In normal hearts, increased stimulation frequency increases force, but in heart failure (HF) this force-frequency relationship (FFR) is often flattened or reversed. Although reduced sarcoplasmic reticulum Ca(2+)-ATPase function may be involved, decreased I(Ca) availability may also contribute. Longer action potential duration (APD), slower intracellular Ca(2+) concentration ([Ca(2+)](i)) decline, and higher diastolic [Ca(2+)](i) in HF could all slow I(Ca) recovery from inactivation, thereby decreasing I(Ca) availability. We measured the effect of different diastolic [Ca(2+)](i) on I(Ca) inactivation and recovery from inactivation in rabbit cardiac myocytes. Both I(Ca) and Ba(2+) current (I(Ba)) were measured. I(Ca) decay was accelerated only at high diastolic [Ca(2+)](i) (600 nM). I(Ba) inactivation was slower but insensitive to [Ca(2+)](i). Membrane potential dependence of I(Ca) or I(Ba) availability was not affected by [Ca(2+)](i) <600 nM. Recovery from inactivation was slowed by both depolarization and high [Ca(2+)](i). We also used perforated patch with action potential (AP)-clamp and normal Ca(2+) transients, using various APDs as conditioning pulses for different frequencies (and to simulate HF APD). Recovery of I(Ca) following longer APD was increasingly incomplete, decreasing I(Ca) availability. Trains of long APs caused a larger I(Ca) decrease than short APD at the same frequency. This effect on I(Ca) availability was exacerbated by slowing twitch [Ca(2+)](i) decline by approximately 50%. We conclude that long APD and slower [Ca(2+)](i) decline lead to cumulative inactivation limiting I(Ca) at high heart rates and might contribute to the negative FFR in HF, independent of altered Ca(2+) channel properties. PMID- 17400725 TI - Cerebrovascular effects of ischemic preconditioning: endothelial survivin joins the fray. PMID- 17400726 TI - SCEPTRANS: an online tool for analyzing periodic transcription in yeast. AB - SUMMARY: SCEPTRANS is designed for analysis of microarray timecourse data related to periodic phenomena in the budding yeast. The server allows for easy viewing of temporal profiles of multiple genes in a number of datasets. Additional functionality includes searching for coexpressed genes, periodicity and correlation analysis, integrating functional annotation and localization data as well as advanced operations on sets of genes. AVAILABILITY: Available online at http://sceptrans.org/ PMID- 17400727 TI - PROTMAP2D: visualization, comparison and analysis of 2D maps of protein structure. AB - MOTIVATION: Protein structure comparison is a fundamental problem in structural biology and bioinformatics. Two-dimensional maps of distances between residues in the structure contain sufficient information to restore the 3D representation, while maps of contacts reveal characteristic patterns of interactions between secondary and super-secondary structures and are very attractive for visual analysis. The overlap of 2D maps of two structures can be easily calculated, providing a sensitive measure of protein structure similarity. PROTMAP2D is a software tool for calculation of contact and distance maps based on user-defined criteria, quantitative comparison of pairs or series of contact maps (e.g. alternative models of the same protein, model versus native structure, different trajectories from molecular dynamics simulations, etc.) and visualization of the results. AVAILABILITY: PROTMAP2D for Windows / Linux / MacOSX is freely available for academic users from http://genesilico.pl/protmap2d.htm PMID- 17400728 TI - Management, presentation and interpretation of genome scans using GSCANDB. AB - MOTIVATION: Advances in high-throughput genotyping have made it possible to carry out genome-wide association studies using very high densities of genetic markers. This has led to the problem of the storage, management, quality control, presentation and interpretation of results. In order to achieve a successful outcome, it may be necessary to analyse the data in different ways and compare the results with genome annotations and other genome scans. RESULTS: We created GSCANDB, a database for genome scan data, using a MySQL backend and Perl-CGI web interface. It displays genome scans of multiple phenotypes analysed in different ways and projected onto genome annotations derived from EnsMart. The current version is optimized for analysis of mouse data, but is customizable to other species. AVAILABILITY: Source code and example data are available under the GPL, in versions tailored to either human or mouse association studies, from http://gscan.well.ox.ac.uk/software. PMID- 17400729 TI - Pulmonary inflammation and emphysema: role of the cytokines IL-18 and IL-13. AB - RATIONALE: Chronic obstructive pulmonary disease (COPD) is believed to be an inflammatory cytokine-driven disease, but a causal basis that can be associated with a specific cytokine has not been directly demonstrated. We have previously reported that proinflammatory cytokine IL-18 expression is important in the pathogenesis of pulmonary inflammation and lung injury in mice. Our results demonstrate that IL-18 overproduction in the lungs can induce lung diseases, such as pulmonary inflammation, lung fibrosis, and COPD. OBJECTIVES: We analyzed the role of IL-18 in the pathogenesis of COPD. METHODS: Using the human surfactant protein C promoter to drive expression of mature mouse IL-18 cDNA, we developed two different lines of transgenic (Tg) mice that overproduced mouse mature IL-18 in the lungs either constitutively or in response to doxycycline. MEASUREMENTS AND MAIN RESULTS: Constitutive overproduction of IL-18 in the lungs resulted in the increased production of IFN-gamma, IL-5, and IL-13, and chronic pulmonary lung inflammation with the appearance of CD8+ T cells, macrophages, neutrophils, and eosinophils. Increased lung volume, severe emphysematous change, dilatation of the right ventricle, and mild pulmonary hypertension were observed in (more than 15-wk-old) Tg mice. Interestingly, disruption of the IL-13 gene, but not the IFN-gamma gene, prevented emphysema and pulmonary inflammation in Tg mice. Moreover, when IL-18 production was induced in lung tissues for 4 weeks through the use of a doxycycline-dependent surfactant protein C promoter, interstitial inflammation was induced. CONCLUSIONS: Our results indicate that IL-18 and IL-13 may have an important role in the pathogenesis of COPD. PMID- 17400730 TI - Inhaled corticosteroid use in chronic obstructive pulmonary disease and the risk of hospitalization for pneumonia. AB - RATIONALE: Inhaled corticosteroids are commonly prescribed to patients with chronic obstructive pulmonary disease (COPD). OBJECTIVES: To examine whether these medications might be associated with an excess risk of pneumonia. METHODS: We conducted a nested case-control study within a cohort of patients with COPD from Quebec, Canada, over the period 1988-2003, identified on the basis of administrative databases linking hospitalization and drug-dispensing information. Each subject hospitalized for pneumonia during follow-up (case subjects) was age and time matched to four control subjects. The effect of the use of inhaled corticosteroids was assessed by conditional logistic regression, after adjusting for comorbidity and COPD severity. MEASUREMENTS AND MAIN RESULTS: The cohort included 175,906 patients with COPD of whom 23,942 were hospitalized for pneumonia during follow-up, for a rate of 1.9 per 100 per year, and matched to 95,768 control subjects. The adjusted rate ratio of hospitalization for pneumonia associated with current use of inhaled corticosteroids was 1.70 (95% confidence interval [CI], 1.63-1.77) and 1.53 (95% CI, 1.30-1.80) for pneumonia hospitalization followed by death within 30 days. The rate ratio of hospitalization for pneumonia was greatest with the highest doses of inhaled corticosteroids, equivalent to fluticasone at 1,000 microg/day or more (rate ratio, 2.25; 95% CI, 2.07-2.44). All-cause mortality was similar for patients hospitalized for pneumonia, whether or not they had received inhaled corticosteroids in the recent past (7.4 and 8.2%, respectively). CONCLUSIONS: The use of inhaled corticosteroids is associated with an excess risk of pneumonia hospitalization and of pneumonia hospitalization followed by death within 30 days, among elderly patients with COPD. PMID- 17400731 TI - C-reactive protein, obstructive sleep apnea, and cognitive dysfunction in school aged children. AB - RATIONALE: Obstructive sleep apnea (OSA) in children is associated with substantial neurobehavioral and cognitive dysfunction. However, not all children with OSA exhibit altered cognitive performance. OBJECTIVES: To assess the magnitude of the systemic inflammatory response, as measured by high-sensitivity C-reactive protein (hsCRP) serum levels which may identify children with OSA at higher susceptibility for cognitive morbidity. METHODS: Habitually snoring children and nonsnoring children (total, 278; age range, 5-7 yr) were recruited from the community, and underwent overnight polysomnography and neurocognitive testing and a blood draw the next morning. Snoring children were divided into OSA and no-OSA groups, and children with OSA were further subdivided into those with two or more abnormal cognitive subtests and into those with normal cognitive scores. Serum levels of hsCRP were also measured. MEASUREMENTS AND MAIN RESULTS: Among snoring children without OSA, mean hsCRP was 0.19+/-0.07 mg/dl compared with 0.36+/-0.11 mg/dl in those with OSA (p<0.01). Furthermore, hsCRP was 0.48+/ 0.12 mg/dl in children with OSA and cognitive deficits, compared with 0.21+/-0.08 mg/dl in children with OSA and normal cognitive scores (p<0.002). CONCLUSIONS: hsCRP levels are higher in children with OSA, and particularly in those who develop neurocognitive deficits, suggesting that the magnitude of the inflammatory responses elicited by OSA is a major determinant of increased risk for neurocognitive dysfunction. PMID- 17400732 TI - Chronic asthma-induced airway remodeling is prevented by toll-like receptor-7/8 ligand S28463. AB - RATIONALE: Allergic asthma is a heterogeneous disease, the pathology of which is a result of improper immune responses to innocuous antigens. We and others have previously shown that one of the Toll-like receptor (TLR)-7/8 ligands, the synthetic compound S28463 (resiquimod, R-848), is able to inhibit acute allergic asthma in mice. OBJECTIVES: Given that the efficiency of this pharmacologic compound against the smooth muscle mass increase and goblet cell hyperplasia that are characteristic of chronic allergic asthma has not been previously assessed, we investigated the ability of this compound to prevent these aspects of chronic airway remodeling. METHODS: The impact of S28463 treatment was assessed in a Brown Norway rat model of chronic asthma by histologic, morphometric, and molecular techniques. MEASUREMENTS AND MAIN RESULTS: We demonstrate that treatment with S28463 is able to prevent the development of goblet cell hyperplasia and increases in airway smooth muscle mass, and that this effect is at least partially mediated by inhibiting proliferation of goblet and smooth muscle cells, respectively. Furthermore, we show that the abrogation of airway remodeling is preceded by inhibition of the inflammatory reaction normally occurring in response to allergen challenge in sensitized animals. This inhibition was associated with a reduction of both helper T cell type 1 and type 2 cytokine protein expression in the lungs, demonstrating the potent antiinflammatory effect of this pharmaceutical compound in the context of allergic reactions. CONCLUSIONS: Taken together, our results indicate great potential for the use of S28463 as an antiinflammatory therapeutic agent for the management of chronic asthma. PMID- 17400733 TI - Complement C3a regulates Muc5ac expression by airway Clara cells independently of Th2 responses. AB - RATIONALE: The factors that control the secretion of epithelial mucins are essential to understanding obstructive airway diseases such as asthma. Although the complement anaphylatoxin C3a and its receptor have been shown to promote many features of allergic lung inflammation, the contribution to mucin expression has not been elucidated. OBJECTIVES: To determine if the C3a receptor with its ligand regulates airway epithelial mucin production. METHODS: Mice deficient in the C3a receptor were examined in a model of allergic airway disease for the presence of goblet cells and the gel-forming secreted mucin Muc5ac. MEASUREMENTS AND MAIN RESULTS: Lungs from antigen-challenged C3a receptor-deficient mice revealed a dramatic decrease in goblet cells and Muc5ac compared with challenged wild-type control animals. These differences were dependent on C3a binding to its receptor since intranasal challenge with C3a induced the formation of goblet cells only in wild-type but not C3a receptor-deficient mice. Increased numbers of goblet cells were also found in C3a-stimulated RAG-1-deficient mice demonstrating a mechanism independent of T lymphocytes and Th2 cytokines, mediators which have been shown to regulate mucin expression. A direct physiological role for C3a in these models was further demonstrated in cultures of airway epithelial Clara cells, which not only express the C3a receptor but also produce Muc5ac in response to C3a. CONCLUSIONS: These studies identify a novel C3a receptor-dependent mechanism in the development of airway epithelial goblet cells and regulation of Muc5ac production and implicate C3a as a mediator of airway obstruction in asthma. PMID- 17400734 TI - The anaerobic regulatory network required for Pseudomonas aeruginosa nitrate respiration. AB - In Pseudomonas aeruginosa, the narK(1)K(2)GHJI operon encodes two nitrate/nitrite transporters and the dissimilatory nitrate reductase. The narK(1) promoter is anaerobically induced in the presence of nitrate by the dual activity of the oxygen regulator Anr and the N-oxide regulator Dnr in cooperation with the nitrate-responsive two-component regulatory system NarXL. The DNA bending protein IHF is essential for this process. Similarly, narXL gene transcription is enhanced under anaerobic conditions by Anr and Dnr. Furthermore, Anr and NarXL induce expression of the N-oxide regulator gene dnr. Finally, NarXL in cooperation with Dnr is required for anaerobic nitrite reductase regulatory gene nirQ transcription. A cascade regulatory model for the fine-tuned genetic response of P. aeruginosa to anaerobic growth conditions in the presence of nitrate was deduced. PMID- 17400735 TI - Membrane-bound nitrate reductase is required for anaerobic growth in cystic fibrosis sputum. AB - The autosomal recessive disorder cystic fibrosis (CF) affects approximately 70,000 people worldwide and is characterized by chronic bacterial lung infections with the opportunistic pathogen Pseudomonas aeruginosa. To form a chronic CF lung infection, P. aeruginosa must grow and proliferate within the CF lung, and the highly viscous sputum within the CF lung provides a likely growth substrate. Recent evidence indicates that anaerobic microenvironments may be present in the CF lung sputum layer. Since anaerobic growth significantly enhances P. aeruginosa biofilm formation and antibiotic resistance, it is important to examine P. aeruginosa physiology and metabolism in anaerobic environments. Measurement of nitrate levels revealed that CF sputum contains sufficient nitrate to support significant P. aeruginosa growth anaerobically, and mutational analysis revealed that the membrane-bound nitrate reductase is essential for P. aeruginosa anaerobic growth in an in vitro CF sputum medium. In addition, expression of genes coding for the membrane-bound nitrate reductase complex is responsive to CF sputum nitrate levels. These findings suggest that the membrane-bound nitrate reductase is critical for P. aeruginosa anaerobic growth with nitrate in the CF lung. PMID- 17400736 TI - Zinc-responsive regulation of alternative ribosomal protein genes in Streptomyces coelicolor involves zur and sigmaR. AB - Streptomyces coelicolor contains paralogous versions of seven ribosomal proteins (S14, S18, L28, L31, L32, L33, and L36), which differ in their potential to bind structural zinc. The paralogues are termed C(+) or C(-) on the basis of the presence or absence of putative cysteine ligands. Here, mutational studies suggest that the C(-) version of L31 can functionally replace its C(+) paralogue only when expressed at an artificially elevated level. We show that the level of expression of four transcriptional units encoding C(-) proteins is elevated under conditions of zinc deprivation. Zur controls the expression of three transcriptional units (including rpmG2, rpmE2, rpmB2, rpsN2, rpmF2, and possibly rpsR2). Zur also controls the expression of the znuACB operon, which is predicted to encode a high-affinity zinc transport system. Surprisingly, the zinc responsive control of the rpmG3-rpmJ2 operon is dictated by sigma(R), a sigma factor that was previously shown to control the response to disulfide stress in S. coelicolor. The induction of sigma(R) activity during zinc limitation establishes an important link between thiol-disulfide metabolism and zinc homeostasis. PMID- 17400737 TI - Genome sequence and analysis of a Propionibacterium acnes bacteriophage. AB - Cutaneous propionibacteria are important commensals of human skin and are implicated in a wide range of opportunistic infections. Propionibacterium acnes is also associated with inflammatory acne vulgaris. Bacteriophage PA6 is the first phage of P. acnes to be sequenced and demonstrates a high degree of similarity to many mycobacteriophages both morphologically and genetically. PA6 possesses an icosahedreal head and long noncontractile tail characteristic of the Siphoviridae. The overall genome organization of PA6 resembled that of the temperate mycobacteriophages, although the genome was much smaller, 29,739 bp (48 predicted genes), compared to, for example, 50,550 bp (86 predicted genes) for the Bxb1 genome. PA6 infected only P. acnes and produced clear plaques with turbid centers, but it lacked any obvious genes for lysogeny. The host range of PA6 was restricted to P. acnes, but the phage was able to infect and lyse all P. acnes isolates tested. Sequencing of the PA6 genome makes an important contribution to the study of phage evolution and propionibacterial genetics. PMID- 17400738 TI - Identification of an AU-rich translational enhancer within the Escherichia coli fepB leader RNA. AB - The fepB gene encodes a periplasmic binding protein that is essential for the uptake of ferric enterobactin by Escherichia coli. Its transcription is regulated in response to iron levels by the Fur repressor. The fepB transcript includes a 217-nucleotide leader sequence with several features suggestive of posttranscriptional regulation. To investigate the fepB leader for its contribution to fepB expression, defined deletions and substitution mutations in the leader were characterized using fepB-phoA translational fusions. The fepB leader was found to be necessary for maximal fepB expression, primarily due to the influence of an AU-rich translational enhancer (TE) located 5' to the Shine Dalgarno sequence. Deletions or substitutions within the TE sequence decreased fepB-phoA expression fivefold. RNase protection and in vitro transcription translation assays demonstrated that the TE augmented translational efficiency, as well as RNA levels. Moreover, primer extension inhibition assays showed that the TE increases ribosome binding. In contrast to the enhancing effect of the TE, the natural fepB GUG start codon decreased ribosome binding and reduced fepB expression 2.5-fold compared with the results obtained with leaders bearing an AUG initiation codon. Thus, the TE-GUG organization in fepB results in an intermediate level of expression compared to the level with AUG, with or without the TE. Furthermore, we found that the TE-GUG sequence is conserved among the eight gram-negative strains examined that have fepB genes, suggesting that this organization may provide a selective advantage. PMID- 17400740 TI - YhdJ, a nonessential CcrM-like DNA methyltransferase of Escherichia coli and Salmonella enterica. AB - The Caulobacter crescentus DNA adenine methyltransferase CcrM and its homologs in the alpha-Proteobacteria are essential for viability. CcrM is 34% identical to the yhdJ gene products of Escherichia coli and Salmonella enterica. This study provides evidence that the E. coli yhdJ gene encodes a DNA adenine methyltransferase. In contrast to an earlier report, however, we show that yhdJ is not an essential gene in either E. coli or S. enterica. PMID- 17400739 TI - Growth rate toxicity phenotypes and homeostatic supercoil control differentiate Escherichia coli from Salmonella enterica serovar Typhimurium. AB - Escherichia coli and Salmonella enterica serovar Typhimurium share high degrees of DNA and amino acid identity for 65% of the homologous genes shared by the two genomes. Yet, there are different phenotypes for null mutants in several genes that contribute to DNA condensation and nucleoid formation. The mutant R436-S form of the GyrB protein has a temperature-sensitive phenotype in Salmonella, showing disruption of supercoiling near the terminus and replicon failure at 42 degrees C. But this mutation in E. coli is lethal at the permissive temperature. A unifying hypothesis for why the same mutation in highly conserved homologous genes of different species leads to different physiologies focuses on homeotic supercoil control. During rapid growth in mid-log phase, E. coli generates 15% more negative supercoils in pBR322 DNA than Salmonella. Differences in compaction and torsional strain on chromosomal DNA explain a complex set of single-gene phenotypes and provide insight into how supercoiling may modulate epigenetic effects on chromosome structure and function and on prophage behavior in vivo. PMID- 17400741 TI - Selectivity for D-lactate incorporation into the peptidoglycan precursors of Lactobacillus plantarum: role of Aad, a VanX-like D-alanyl-D-alanine dipeptidase. AB - Lactobacillus plantarum produces peptidoglycan precursors ending in D-lactate instead of D-alanine, making the bacterium intrinsically resistant to vancomycin. The ligase Ddl of L. plantarum plays a central role in this specificity by synthesizing D-alanyl-D-lactate depsipeptides that are added to the precursor peptide chain by the enzyme MurF. Here we show that L. plantarum also encodes a D Ala-D-Ala dipeptidase, Aad, which eliminates D-alanyl-D-alanine dipeptides that are produced by the Ddl ligase, thereby preventing their incorporation into the precursors. Although D-alanine-ended precursors can be incorporated into the cell wall, inactivation of Aad failed to suppress growth defects of L. plantarum mutants deficient in d-lactate-ended precursor synthesis. PMID- 17400742 TI - Re-citrate synthase from Clostridium kluyveri is phylogenetically related to homocitrate synthase and isopropylmalate synthase rather than to Si-citrate synthase. AB - The synthesis of citrate from acetyl-coenzyme A and oxaloacetate is catalyzed in most organisms by a Si-citrate synthase, which is Si-face stereospecific with respect to C-2 of oxaloacetate. However, in Clostridium kluyveri and some other strictly anaerobic bacteria, the reaction is catalyzed by a Re-citrate synthase, whose primary structure has remained elusive. We report here that Re-citrate synthase from C. kluyveri is the product of a gene predicted to encode isopropylmalate synthase. C. kluyveri is also shown to contain a gene for Si citrate synthase, which explains why cell extracts of the organism always exhibit some Si-citrate synthase activity. PMID- 17400743 TI - Analysis of LuxR regulon gene expression during quorum sensing in Vibrio fischeri. AB - The regulation of the lux operon (luxICDABEG) of Vibrio fischeri has been intensively studied as a model for quorum sensing in proteobacteria. Two dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis previously identified several non-Lux proteins in V. fischeri MJ-100 whose expression was dependent on LuxR and 3-oxo-hexanoyl-l-homoserine lactone (3-oxo C6-HSL). To determine if the LuxR-dependent regulation of the genes encoding these proteins was due to direct transcriptional control by LuxR and 3-oxo-C6-HSL or instead was due to indirect control via an unidentified regulatory element, promoters of interest were cloned into a lacZ reporter and tested for their LuxR and 3-oxo-C6-HSL dependence in recombinant Escherichia coli. The promoters for qsrP, acfA, and ribB were found to be directly activated via LuxR-3-oxo-C6-HSL. The sites of transcription initiation were established via primer extension analysis. Based on this information and the position of the lux box-binding site near position -40, all three promoters appear to have a class II-type promoter structure. In order to more fully characterize the LuxR regulon in V. fischeri MJ 100, real-time reverse transcription-PCR was used to study the temporal expression of qsrP, acfA, and ribB during the exponential and stationary phases of growth, and electrophoretic mobility shift assays were used to compare the binding affinities of LuxR to the promoters under investigation. Taken together, the results demonstrate that regulation of the production of QsrP, RibB, and AcfA is controlled directly by LuxR at the level of transcription, thereby establishing that there is a LuxR regulon in V. fischeri MJ-100 whose genes are coordinately expressed during mid-exponential growth. PMID- 17400744 TI - ScrG, a GGDEF-EAL protein, participates in regulating swarming and sticking in Vibrio parahaemolyticus. AB - In this work, we describe a new gene controlling lateral flagellar gene expression. The gene encodes ScrG, a protein containing GGDEF and EAL domains. This is the second GGDEF-EAL-encoding locus determined to be involved in the regulation of swarming: the first was previously characterized and named scrABC (for "swarming and capsular polysaccharide regulation"). GGDEF and EAL domain containing proteins participate in the synthesis and degradation of the nucleotide signal cyclic di-GMP (c-di-GMP) in many bacteria. Overexpression of scrG was sufficient to induce lateral flagellar gene expression in liquid, decrease biofilm formation, decrease cps gene expression, and suppress the DeltascrABC phenotype. Removal of its EAL domain reversed ScrG activity, converting ScrG to an inhibitor of swarming and activator of cps expression. Overexpression of scrG decreased the intensity of a (32)P-labeled nucleotide spot comigrating with c-di-GMP standard, whereas overexpression of scrG(Delta)(EAL) enhanced the intensity of the spot. Mutants with defects in scrG showed altered swarming and lateral flagellin production and colony morphology (but not swimming motility); furthermore, mutation of two GGDEF-EAL-encoding loci (scrG and scrABC) produced cumulative effects on swarming, lateral flagellar gene expression, lateral flagellin production and colony morphology. Mutant analysis supports the assignment of the primary in vivo activity of ScrG to acting as a phosphodiesterase. The data are consistent with a model in which multiple GGDEF EAL proteins can influence the cellular nucleotide pool: a low concentration of c di-GMP favors surface mobility, whereas high levels of this nucleotide promote a more adhesive Vibrio parahaemolyticus cell type. PMID- 17400745 TI - An extracytoplasmic function sigma factor acts as a general stress response regulator in Sinorhizobium meliloti. AB - Sinorhizobium meliloti genes transcriptionally up-regulated after heat stress, as well as upon entry into stationary phase, were identified by microarray analyses. Sixty stress response genes were thus found to be up-regulated under both conditions. One of them, rpoE2 (smc01506), encodes a putative extracytoplasmic function (ECF) sigma factor. We showed that this sigma factor controls its own transcription and is activated by various stress conditions, including heat and salt, as well as entry into stationary phase after either carbon or nitrogen starvation. We also present evidence that the product of the gene cotranscribed with rpoE2 negatively regulates RpoE2 activity, and we therefore propose that it plays the function of anti-sigma factor. By combining transcriptomic, bioinformatic, and quantitative reverse transcription-PCR analyses, we identified 44 RpoE2-controlled genes and predicted the number of RpoE2 targets to be higher. Strikingly, more than one-third of the 60 stress response genes identified in this study are RpoE2 targets. Interestingly, two genes encoding proteins with known functions in stress responses, namely, katC and rpoH2, as well as a second ECF-encoding gene, rpoE5, were found to be RpoE2 regulated. Altogether, these data suggest that RpoE2 is a major global regulator of the general stress response in S. meliloti. Despite these observations, and although this sigma factor is well conserved among alphaproteobacteria, no in vitro nor in planta phenotypic difference from the wild-type strain could be detected for rpoE2 mutants. This therefore suggests that other important actors in the general stress response have still to be identified in S. meliloti. PMID- 17400746 TI - Spontaneous trpY mutants and mutational analysis of the TrpY archaeal transcription regulator. AB - Over 90% of Methanothermobacter thermautotrophicus mutants isolated as spontaneously resistant to 5-methyl tryptophan had mutations in trpY. Most were single-base-pair substitutions that identified separate DNA- and tryptophan binding regions in TrpY. In vivo and in vitro studies revealed that DNA binding was sufficient for TrpY repression of trpY transcription but that TrpY must bind DNA and tryptophan to assemble a complex that represses trpEGCFBAD. PMID- 17400747 TI - Microcin J25 has dual and independent mechanisms of action in Escherichia coli: RNA polymerase inhibition and increased superoxide production. AB - Microcin J25 (MccJ25) uptake by Escherichia coli requires the outer membrane receptor FhuA and the inner membrane proteins TonB, ExbD, ExbB, and SbmA. MccJ25 appears to have two intracellular targets: (i) RNA polymerase (RNAP), which has been described in E. coli and Salmonella enterica serovars, and (ii) the respiratory chain, reported only in S. enterica serovars. In the current study, it is shown that the observed difference between the actions of microcin on the respiratory chain in E. coli and S. enterica is due to the relatively low microcin uptake via the chromosomally encoded FhuA. Higher expression by a plasmid-encoded FhuA allowed greater uptake of MccJ25 by E. coli strains and the consequent inhibition of oxygen consumption. The two mechanisms, inhibition of RNAP and oxygen consumption, are independent of each other. Further analysis revealed for the first time that MccJ25 stimulates the production of reactive oxygen species (O(2)(*-)) in bacterial cells, which could be the main reason for the damage produced on the membrane respiratory chain. PMID- 17400748 TI - Insights into the autotrophic CO2 fixation pathway of the archaeon Ignicoccus hospitalis: comprehensive analysis of the central carbon metabolism. AB - Ignicoccus hospitalis is an autotrophic hyperthermophilic archaeon that serves as a host for another parasitic/symbiotic archaeon, Nanoarchaeum equitans. In this study, the biosynthetic pathways of I. hospitalis were investigated by in vitro enzymatic analyses, in vivo (13)C-labeling experiments, and genomic analyses. Our results suggest the operation of a so far unknown pathway of autotrophic CO(2) fixation that starts from acetyl-coenzyme A (CoA). The cyclic regeneration of acetyl-CoA, the primary CO(2) acceptor molecule, has not been clarified yet. In essence, acetyl-CoA is converted into pyruvate via reductive carboxylation by pyruvate-ferredoxin oxidoreductase. Pyruvate-water dikinase converts pyruvate into phosphoenolpyruvate (PEP), which is carboxylated to oxaloacetate by PEP carboxylase. An incomplete citric acid cycle is operating: citrate is synthesized from oxaloacetate and acetyl-CoA by a (re)-specific citrate synthase, whereas a 2 oxoglutarate-oxidizing enzyme is lacking. Further investigations revealed that several special biosynthetic pathways that have recently been described for various archaea are operating. Isoleucine is synthesized via the uncommon citramalate pathway and lysine via the alpha-aminoadipate pathway. Gluconeogenesis is achieved via a reverse Embden-Meyerhof pathway using a novel type of fructose 1,6-bisphosphate aldolase. Pentosephosphates are formed from hexosephosphates via the suggested ribulose-monophosphate pathway, whereby formaldehyde is released from C-1 of hexose. The organism may not contain any sugar-metabolizing pathway. This comprehensive analysis of the central carbon metabolism of I. hospitalis revealed further evidence for the unexpected and unexplored diversity of metabolic pathways within the (hyperthermophilic) archaea. PMID- 17400749 TI - Septic arthritis of the knee after anterior cruciate ligament surgery: a stage adapted treatment regimen. AB - BACKGROUND: Septic arthritis is a rare complication after anterior cruciate ligament surgery. All case series reported so far have been retrospective, and case numbers of septic arthritis have ranged from 4 to 11. HYPOTHESIS: A stage adapted treatment regimen for septic arthritis after anterior cruciate ligament surgery can provide reliable results. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: From June 1993 to May 2003, 24 patients met the inclusion criteria for this study. The average age at trauma was 32.5 years. Treatment protocol was based on the grade of infection. Options included transarthroscopic treatment for Gaechter grades 1 and 2 infections or medial and lateral arthrotomy for grades 3 and 4 infections. Graft retention was decided based on clinical findings. The setting was a specialized trauma hospital. Follow-up included International Knee Documentation Committee forms, radiographs, and the Tegner and Lysholm scores at a mean of 66 months (range, 11-142) after treatment. RESULTS: In all cases, treatment of infection was successful; overall, a mean of 2.2 operations were required. In 7 cases, it was possible to salvage the graft. The Tegner activity level before the knee injury was 6.1 points. At follow-up, the average score was 3.8 points. The postoperative subjective International Knee Documentation Committee score averaged 64. The mean Lysholm score was 65.6. On clinical examination, a mean extension deficit of 3 degrees and a mean maximum flexion of 120 degrees were found. In the single-legged hop test, a mean capacity of 63% compared with the uninjured side was measured. CONCLUSION: The described treatment regimen gives reliable results for this complication. There were no recurrences of septic arthritis or bone infection. Early infection can be managed arthroscopically with satisfactory results regarding the treatment of infection. In advanced or chronic infection, a more radical approach seems favorable. Results in these cases are overall only fair compared with patients with an uncomplicated anterior cruciate ligament reconstruction. PMID- 17400750 TI - An analysis of soft tissue allograft anterior cruciate ligament reconstruction in a rabbit model: a short-term study of the use of mesenchymal stem cells to enhance tendon osteointegration. AB - BACKGROUND: Soft tissue allografts are essential for revision and multiple ligament surgeries in the knee, where donor-site morbidity is an issue. However, the use of allografts is associated with a higher failure rate of osteointegration. Mesenchymal stem cells (MSCs) are investigated as potential agents to enhance bone tunnel and tendon healing. PURPOSE: This study was conducted to analyze the effect of coating allografts with MSCs on the quality and rate of osteointegration at the allograft tendon and bone interface, and the biomechanical properties of these enhanced anterior cruciate ligament (ACL) grafts compared with controls. STUDY DESIGN: Descriptive laboratory study. METHODS: Bilateral ACL reconstructions using Achilles tendon allografts were performed in 36 rabbits. On 1 limb, the graft was coated with autogenous MSCs in a fibrin glue carrier, while the contralateral limb served as a control with no MSCs. The reconstructions were assessed histologically and biomechanically at 2, 4, and 8 weeks. RESULTS: At 8 weeks, histologic analysis of the controls revealed the development of mature scar tissue resembling Sharpey fibers spanning the tendon-bone interface. In contrast, the MSC-enhanced reconstructions showed a mature zone of fibrocartilage blending from bone to the allograft, strongly resembling a normal ACL insertion. On biomechanical testing, the MSC-enhanced grafts had significantly higher load-to-failure rates than controls. However, the stiffness and Young's modulus were lower in the treatment group. CONCLUSIONS: The application of MSCs at the allograft tendon-bone interface during ACL reconstruction results in the development of an intervening zone of fibrocartilage. The use of MSCs to enhance allograft osteointegration is a novel method offering the potential of more physiologic and earlier healing, although further investigation must be conducted to improve the biomechanical strength. CLINICAL RELEVANCE: Mesenchymal stem cells can improve the biologic properties of soft tissue allograft healing. Combined with the decrease in donor-site morbidity, allografts are a viable choice for the sports medicine surgeon. PMID- 17400751 TI - Inhibition of CFTR Cl- channel function caused by enzymatic hydrolysis of sphingomyelin. AB - Numerous mutations in the cystic fibrosis (CF) transmembrane conductance regulator (CFTR, a Cl(-) channel) disrupt salt and fluid transport and lead to the formation of thick mucus in patients' airways. Obstruction by mucus predisposes CF patients to chronic infections and inflammation, which become gradually harder to control and eventually fatal. Aggressive antibiotic therapy and supportive measures have dramatically lengthened CF patients' lives. Here, we report that sphingomyelinases (SMase) from human respiratory pathogens strongly inhibit CFTR function. The hydrolysis of sphingomyelin by SMase makes it more difficult to activate CFTR by phosphorylation of its regulatory domain. By inhibiting CFTR currents, SMase-producing respiratory tract bacteria may not only aggravate pulmonary infection in some CF patients but may also elicit a condition, analogous to CFTR deficiency, in non-CF patients suffering from bacterial lung infection. PMID- 17400752 TI - Faithful modeling of transient expression and its application to elucidating negative feedback regulation. AB - Modeling and analysis of genetic regulatory networks is essential both for better understanding their dynamic behavior and for elucidating and refining open issues. We hereby present a discrete computational model that effectively describes the transient and sequential expression of a network of genes in a representative developmental pathway. Our model system is a transcriptional cascade that includes positive and negative feedback loops directing the initiation and progression through meiosis in budding yeast. The computational model allows qualitative analysis of the transcription of early meiosis-specific genes, specifically, Ime2 and their master activator, Ime1. The simulations demonstrate a robust transcriptional behavior with respect to the initial levels of Ime1 and Ime2. The computational results were verified experimentally by deleting various genes and by changing initial conditions. The model has a strong predictive aspect, and it provides insights into how to distinguish among and reason about alternative hypotheses concerning the mode by which negative regulation through Ime1 and Ime2 is accomplished. Some predictions were validated experimentally, for instance, showing that the decline in the transcription of IME1 depends on Rpd3, which is recruited by Ime1 to its promoter. Finally, this general model promotes the analysis of systems that are devoid of consistent quantitative data, as is often the case, and it can be easily adapted to other developmental pathways. PMID- 17400753 TI - Killer T cells regulate antigen presentation for early expansion of memory, but not naive, CD8+ T cell. AB - Antigen presentation within the lymph node draining a site of infection is crucial for initiation of cytotoxic T cell responses. Precisely how this antigen presentation regulates T cell expansion in vivo is unclear. Here, we show that, in primary infection, antigen presentation peaks approximately 3 days postinfection and then slowly decays until day 12. This prolonged antigen presentation is required for optimal expansion of naive CD8(+) T cells, because early ablation of dendritic cells reduces the later CD8(+) T cell response. Antigen presentation during secondary infection was 10-fold lower in magnitude and largely terminated by day 4 postinfection. Expansion of memory, but not naive, antigen-specific T cells was tightly controlled by perforin-dependent cytolysis of antigen-presenting cells. The ability of the memory T cells to remove antigen-presenting cells provides a negative-feedback loop to directly limit the duration of antigen presentation in vivo. PMID- 17400754 TI - Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function. AB - The human tripartite motif (TRIM) family comprises 70 members, including HIV restriction factor TRIM5alpha and disease-associated proteins TRIM20 (pyrin) and TRIM21. TRIM proteins have conserved domain architecture but diverse cellular roles. Here, we describe how the C-terminal PRYSPRY domain mediates diverse TRIM functions. The crystal structure of TRIM21 PRYSPRY in complex with its target IgG Fc reveals a canonical binding interface comprised of two discrete pockets formed by antibody-like variable loops. Alanine scanning of this interface has identified the hot-spot residues that control TRIM21 binding to Fc; the same hot spots control HIV/murine leukemia virus restriction by TRIM5alpha and mediate severe familial Mediterranean fever in TRIM20/pyrin. Characterization of the IgG binding site for TRIM21 PRYSPRY reveals TRIM21 as a superantigen analogous to bacterial protein A and suggests that an antibody bipolar bridging mechanism may contribute to the pathogenic accumulation of anti-TRIM21 autoantibody immune complex in autoimmune disease. PMID- 17400755 TI - Connexin30 deficiency causes instrastrial fluid-blood barrier disruption within the cochlear stria vascularis. AB - The endocochlear potential (EP) is essential to hearing, because it provides approximately half of the driving force for the mechanoelectrical transduction current in auditory hair cells. The EP is produced by the stria vascularis (SV), a vascularized bilayer epithelium of the cochlea lateral wall. The absence of the gap junction protein connexin30 (Cx30) in Cx30(-/-) mice results in the SV failure to produce an EP, which mainly accounts for the severe congenital hearing impairment of these mice. Here, we show that the SV components of the EP electrogenic machinery and the epithelial barriers limiting the intrastrial fluid space, which are both necessary for the EP production, were preserved in Cx30(-/ ) mice. In contrast, the endothelial barrier of the capillaries supplying the SV was disrupted before EP onset. This disruption is expected to result in an intrastrial electric shunt that is sufficient to account for the absence of the EP production. Immunofluorescence analysis of wild-type mice detected Cx30 in the basal and intermediate cells of the SV but not in the endothelial cells of the SV capillaries. Moreover, dye-coupling experiments showed that endothelial cells were not coupled to the SV basal, intermediate, and marginal cells. SV transcriptome analysis revealed a significant down-regulation of betaine homocysteine S-methyltransferase (Bhmt) in the Cx30(-/-) mice, which was restricted to the SV and resulted in a local increase in homocysteine, a known factor of endothelial dysfunction. Disruption of the SV endothelial barrier is a previously undescribed pathogenic process underlying hearing impairment. PMID- 17400756 TI - Primary patency with cutting and conventional balloon angioplasty for different types of hemodialysis access stenosis. AB - PURPOSE: To compare primary patency rates of cutting balloon percutaneous transluminal angioplasty (PTA) (hereafter, cutting PTA) and conventional balloon PTA (hereafter, conventional PTA) in the treatment of different types of hemodialysis access stenosis. MATERIALS AND METHODS: The institutional review board approved this study. Written informed consent was obtained for the prospective component of this study and waived for the retrospective component. Patients in whom treatment with cutting PTA alone or conventional PTA alone was clinically successful formed the two study groups. Primary patency for the lesion was defined as uninterrupted patency of the treated site after balloon PTA. A site was no longer considered patent when the patient underwent treatment for hemodialysis access failure due to restenosis of the treated site. Primary patency rates for lesions were calculated with the Kaplan-Meier method according to the type of stenosis. We compared the two groups by using the log-rank test to determine statistical significance. RESULTS: In the cutting PTA group, 62 patients with 77 stenoses (32 men, 30 women; mean age, 65.5 years +/- 10.1 [standard deviation]) achieved clinical success. In the conventional PTA group, 52 patients with 68 stenoses (23 men, 29 women; mean age, 61.9 years +/- 10.2) achieved clinical success. In patients with autogenous venous stenosis, no significant difference in the primary patency rate was noted between groups (P = .369). In patients with graft-to-vein anastomotic stenosis, the primary patency rate was significantly higher for cutting PTA than for conventional PTA (P = .39). In patients with intragraft stenosis, no significant difference in the primary patency rate was noted between groups (P = .379). In patients with in stent restenosis, no significant difference in the primary patency rate was noted between groups (P = .923). CONCLUSION: Primary patency rates are significantly higher for cutting PTA in the treatment of graft-to-vein anastomotic stenosis; however, no significant differences in primary patency rates exist between these PTAs in the treatment of autogenous venous stenosis, intragraft stenosis, or in stent restenosis. PMID- 17400757 TI - Lung carcinoma: diffusion-weighted mr imaging--preliminary evaluation with apparent diffusion coefficient. AB - PURPOSE: To prospectively evaluate diffusion-weighted (DW) magnetic resonance (MR) imaging with a split acquisition of fast spin-echo signals for diffusion imaging (SPLICE) sequence for tissue characterization of lung carcinomas by using apparent diffusion coefficients (ADCs). MATERIALS AND METHODS: An institutional review board approved this study; informed consent was obtained from patients. Thirty patients (nine women, 21 men; mean age, 68.0 years) with lung carcinoma underwent DW MR imaging with the SPLICE sequence. ADC of each lung carcinoma was calculated from DW MR images obtained with low and high b values. ADCs of lung carcinomas were statistically compared among histologic types. Nine surgically excised lung carcinomas were evaluated for correlation between ADCs and tumor cellularities. Analysis of variance was used to determine changes in ADCs and histologic lung carcinoma types. Spearman rank correlation was calculated between ADCs and tumor cellularities. RESULTS: ADCs for lung carcinomas were 1.63 x 10( 3) mm(2)/sec +/- 0.5 (mean +/- standard deviation) for squamous cell carcinoma, 2.12 x 10(-3) mm(2)/sec +/- 0.6 for adenocarcinoma, 1.30 x 10(-3) mm(2)/sec +/- 0.4 for large-cell carcinoma, and 2.09 x 10(-3) mm(2)/sec +/- 0.3 for small-cell carcinoma. ADC of adenocarcinoma was significantly higher than that of squamous cell carcinoma and large-cell carcinoma (P < .05). ADCs were 1.59 x 10(-3) mm(2)/sec +/- 0.5 and 1.70 x 10(-3) mm(2)/sec +/- 0.4 for moderately and poorly differentiated squamous cell carcinoma, respectively. ADCs were 2.52 x 10(-3) mm(2)/sec +/- 0.4 and 1.44 x 10(-3) mm(2)/sec +/- 0.3 for well- and poorly differentiated adenocarcinoma, respectively. ADC of well-differentiated adenocarcinoma was significantly higher than that of moderately and poorly differentiated squamous cell carcinoma and poorly differentiated adenocarcinoma (P < .05). With the Spearman rank test, ADCs of lung carcinomas correlated well with tumor cellularities (Spearman coefficient, -0.75; P < .02). CONCLUSION: ADCs of lung carcinomas overlap, but ADCs of well-differentiated adenocarcinoma appear to be higher than those of other histologic lung carcinoma types. PMID- 17400758 TI - Ischemic brain tissue water content: CT monitoring during middle cerebral artery occlusion and reperfusion in rats. AB - PURPOSE: To prospectively perform computed tomography (CT) in rats to determine whether ischemic edema can be reversed by using early arterial reperfusion. MATERIALS AND METHODS: This study was approved by the local animal protection committee. A suture model was used to occlude the right middle cerebral artery (MCA) in rats for 1, 2, 3, or 4 hours. X-ray attenuation of the brain was measured directly before reperfusion and repeatedly during reperfusion for up to 24 hours. Infarct volumes were determined with triphenyltetrazolium chloride staining. Means of attenuation and infarct volume were compared between hemispheres and groups with a paired t test and analysis of variance. Mixed linear models were applied to compare attenuation among groups over time. RESULTS: During MCA occlusion, attenuation decreased to 69.3 HU +/- 1.9 (standard deviation) after 1 hour (n=12), 66.6 HU +/- 2.0 after 2 hours (n=10), 65.4 HU +/- 2.9 after 3 hours (n=11), and 64.1 HU +/- 1.8 after 4 hours (n=9) (P<.0001). After reperfusion, attenuation remained stable in the 1-hour occlusion group (P=.16) but further and steadily declined in the 2-, 3-, and 4-hour occlusion groups (P<.001). Attenuation during reperfusion in the 1-hour occlusion group differed significantly from that in the 2-, 3-, and 4-hour occlusion groups. CONCLUSION: CT is able to help monitor ischemic edema after MCA occlusion and reperfusion. Ischemic brain edema was not consistently reversible with reperfusion, even after 1 hour of occlusion, and further increased with reperfusion induced at 2 hours or later. PMID- 17400759 TI - Diagnosis of articular cartilage abnormalities of the knee: prospective clinical evaluation of a 3D water-excitation true FISP sequence. AB - PURPOSE: To prospectively evaluate the accuracy of three-dimensional (3D) water excitation true fast imaging with steady-state precession (FISP) in the assessment of cartilage abnormalities of the knee, by using surgery as the reference standard. MATERIALS AND METHODS: The study was approved by the hospital institutional review board. Written informed consent was obtained from all patients. Twenty-nine patients (30 knees) with a mean age of 56 years (range, 18 86 years) were prospectively evaluated with a sagittal 3D true FISP magnetic resonance (MR) sequence. The mean interval between MR imaging and surgery was 1 day (range, 0-9 days). During surgery, the articular surfaces of the knee were evaluated by using a modified Noyes score. The MR images were evaluated by two blinded readers on two separate occasions. Diagnostic performance was evaluated by setting the cutoff for abnormality between grade 1 (intact cartilage surface) and grade 2 (cartilage defects). Statistical methods used included calculation of sensitivity, specificity, and accuracy, with 95% confidence intervals (Wilson score method) and calculation of kappa values with standard errors. RESULTS: Overall sensitivity, specificity, and accuracy for the two readers and the two evaluations ranged from 56% to 66%, 78% to 93%, and 71% to 75%, respectively. Interobserver agreement was substantial for both the first (kappa = 0.73) and the second (kappa = 0.65) evaluation. Intraobserver agreement was almost perfect (kappa = 0.84) for reader 1 and moderate (kappa = 0.60) for reader 2. CONCLUSION: The 3D water-excitation true FISP MR sequence allows assessment of the articular cartilage of the knee with moderate-to-high specificity and low-to-moderate sensitivity. PMID- 17400760 TI - Electromagnetic breast imaging: results of a pilot study in women with abnormal mammograms. AB - PURPOSE: To prospectively assess quantitatively the inherent contrast of electromagnetic (EM) properties that can be imaged by using available technology in women with abnormal findings at conventional breast imaging who underwent subsequent biopsy. MATERIALS AND METHODS: The protocol was HIPAA compliant and approved by the institutional review board. All participants provided informed consent. Fifty-three women with normal (Breast Imaging Reporting and Data System [BI-RADS] category 1) and ninety-seven women with abnormal (BI-RADS category 4 or 5) screening mammograms were imaged with three EM imaging methods: electrical impedance spectroscopy (EIS), microwave imaging spectroscopy (MIS), and near infrared spectral tomography (NIR). A region-of-interest (ROI) analysis was used to assess the EM image properties for comparison of findings with conventional image findings and correlation with specific pathologic parameters for women with abnormal findings. Statistical analyses were conducted. RESULTS: One hundred fifty participants (age range, 35-81 years) were included. EM image property contrast ratios of 150%-200% were found in breast abnormality ROIs relative to the ipsilateral breast background. Analysis of variance demonstrated significant differences in ROI image summaries of mammographically normal versus abnormal breasts for EIS, across diagnostic groups for NIR, and for MIS (analysis restricted to lesions larger than 1 cm(3)). Receiver operating curve (ROC) analysis of the EM properties for cancers among subjects with BI-RADS category 4 or 5, compared with the EM properties for the subjects with normal breasts (BI RADS category 1), yielded areas under the ROC curve ranging from 0.67 to 0.81. Pathologic correlations with mean vessel density, mean vessel area, and epithelium-to-stroma ratio suggest a biological origin of the EM image properties associated with disease. CONCLUSION: Results from EM breast examinations provide statistical evidence of a mean increase in image contrast of 150%-200% between abnormal (benign and malignant) and normal breast tissue. PMID- 17400761 TI - Atypical chest pain: coronary, aortic, and pulmonary vasculature enhancement at biphasic single-injection 64-section CT angiography. AB - PURPOSE: To prospectively evaluate the enhancement of coronary, pulmonary, and thoracic aortic vasculature by using biphasic single-acquisition 64-section computed tomographic (CT) angiography and to prospectively evaluate if differences in right side of the heart and coronary venous enhancement interfere with interpretation of coronary arteries. MATERIALS AND METHODS: With internal review board approval and HIPAA compliance, 50 patients (16 men, 34 women; mean age, 51.5 years; range, 30-75 years) with atypical chest pain were referred from the emergency department and were imaged with a 64-section CT scanner after premedication with oral atenolol and/or intravenous metoprolol. Thoracic CT angiography with retrospective gating was subsequently performed with a single biphasic injection of 130 mL of iso-osmolar contrast material (100 mL at 5 mL/sec and 30 mL at 3 mL/sec) in caudal-to-cranial acquisition. Coronary, aortic, and pulmonary arterial attenuation values were obtained. Coronary venous and right atrial enhancement were evaluated to assess whether there was interference with coronary artery evaluation. A two-tailed Friedman test was used to evaluate differences among segments within each artery. RESULTS: Mean coronary arterial, pulmonary arterial, and aortic attenuation values were significantly higher than the 250-HU threshold (P < .05). Mean pooled coronary arterial (288.9 HU +/- 64.8), pulmonary arterial (316.4 HU +/- 79.9), and aortic (329.9 HU +/- 63.3) attenuation values were significantly higher than the 250-HU threshold (P < .0001). Coronary venous enhancement did not affect depiction or interpretation of coronary arteries. Right atrial streak artifact focally traversed the right coronary artery in only one study. CONCLUSION: The aforementioned thoracic CT angiographic protocol provides enhancement of coronary, aortic, and pulmonary vasculature in a single breath hold without interference from right side of the heart streak artifact or coronary venous enhancement. PMID- 17400762 TI - Glycogen synthase kinase 3 regulates N-methyl-D-aspartate receptor channel trafficking and function in cortical neurons. AB - Emerging evidence has suggested that glycogen synthase kinase 3 (GSK-3) is a key regulatory kinase involved in a plethora of processes in the nervous system, including neuronal development, mood stabilization, and neurodegeneration. However, the cellular mechanisms underlying the actions of GSK-3 remain to be identified. In this study, we examined the impact of GSK-3 on the N-methyl-D aspartate (NMDA) receptor channel, a central player involved in cognitive and emotional processes. We found that application of various structurally different GSK-3 inhibitors caused a long-lasting reduction of NMDA receptor-mediated ionic and synaptic current in cortical pyramidal neurons. Cellular knockdown of GSK 3beta in neuronal cultures with a small interfering RNA led to smaller NMDA receptor current and loss of its regulation by GSK-3 inhibitors. The NR2B subunit containing NMDA receptor was the primary target of GSK-3, but the GSK-3 modulation of NMDAR current did not involve the motor protein kinesin superfamily member 17-based transport of NR2B-containing vesicles along microtubules. Combined electrophysiological, immunocytochemical, and biochemical evidence indicated that GSK-3 inhibitors induced the down-regulation of NMDAR current through increasing the Rab5-mediated and PSD-95-regulated NMDAR internalization in a clathrin/dynamin-dependent manner. PMID- 17400763 TI - Hypoxia inhibits paclitaxel-induced apoptosis through adenosine-mediated phosphorylation of bad in glioblastoma cells. AB - Solid tumors contain hypoxic cells that are resistant to radiotherapy and chemotherapy. The resistance in glioblastoma has been linked to the expression of antiapoptotic Bcl-2 family members. In this study, we found that in human glioblastoma cells hypoxia induces the phosphorylation of the Bcl-2 family protein Bad, thus protecting hypoxic cells from paclitaxel-induced apoptosis. Akt activation is required for the hypoxia-induced protection. In contrast, the extracellular signal-regulated kinase 1/2 activities have only a partial effect, being able to modulate Bad phosphorylation but not paclitaxel-induced apoptosis in hypoxia. We also demonstrated that the degradation of adenosine with adenosine deaminase, the knockdown of A(3) adenosine receptor expression by gene silencing, and the blockade of this receptor through A(3) receptor antagonists blocked the hypoxia-induced phosphorylation of Bad and the prolonged cell survival after treatment with paclitaxel in hypoxia. Thus, the adenosinergic signaling may be an essential component in the hypoxia survival pathway. These results suggest that hypoxia-induced chemoresistance of human glioblastoma cells may occur in a novel mechanism involving activation of adenosine-A(3) receptor-Akt pathway, which mediates Bad inactivation and favors cell survival. PMID- 17400764 TI - Inhibition of transforming growth factor beta signaling reduces pancreatic adenocarcinoma growth and invasiveness. AB - Transforming growth factor beta (TGFbeta) is a pleiotropic factor that regulates cell proliferation, angiogenesis, metastasis, and immune suppression. Dysregulation of the TGFbeta pathway in tumor cells often leads to resistance to the antiproliferative effects of TGFbeta while supporting other cellular processes that promote tumor invasiveness and growth. In the present study, SD 208, a 2,4-disubstituted pteridine, ATP-competitive inhibitor of the TGFbeta receptor I kinase (TGFbetaRI), was used to inhibit cellular activities and tumor progression of PANC-1, a human pancreatic tumor line. SD-208 blocked TGFbeta dependent Smad2 phosphorylation and expression of TGFbeta-inducible proteins in cell culture. cDNA microarray analysis and functional gene clustering identified groups of TGFbeta-regulated genes involved in metastasis, angiogenesis, cell proliferation, survival, and apoptosis. These gene responses were inhibited by SD 208. Using a Boyden chamber motility assay, we demonstrated that SD-208 inhibited TGFbeta-stimulated invasion in vitro. An orthotopic xenograft mouse model revealed that SD-208 reduced primary tumor growth and decreased the incidence of metastasis in vivo. Our findings suggest mechanisms through which TGFbeta signaling may promote tumor progression in pancreatic adenocarcinoma. Moreover, they suggest that inhibition of TGFbetaRI with a small-molecule inhibitor may be effective as a therapeutic approach to treat human pancreatic cancer. PMID- 17400765 TI - Characterization of a gene cluster responsible for the biosynthesis of anticancer agent FK228 in Chromobacterium violaceum No. 968. AB - A gene cluster responsible for the biosynthesis of anticancer agent FK228 has been identified, cloned, and partially characterized in Chromobacterium violaceum no. 968. First, a genome-scanning approach was applied to identify three distinctive C. violaceum no. 968 genomic DNA clones that code for portions of nonribosomal peptide synthetase and polyketide synthase. Next, a gene replacement system developed originally for Pseudomonas aeruginosa was adapted to inactivate the genomic DNA-associated candidate natural product biosynthetic genes in vivo with high efficiency. Inactivation of a nonribosomal peptide synthetase-encoding gene completely abolished FK228 production in mutant strains. Subsequently, the entire FK228 biosynthetic gene cluster was cloned and sequenced. This gene cluster is predicted to encompass a 36.4-kb DNA region that includes 14 genes. The products of nine biosynthetic genes are proposed to constitute an unusual hybrid nonribosomal peptide synthetase-polyketide synthase-nonribosomal peptide synthetase assembly line including accessory activities for the biosynthesis of FK228. In particular, a putative flavin adenine dinucleotide-dependent pyridine nucleotide-disulfide oxidoreductase is proposed to catalyze disulfide bond formation between two sulfhydryl groups of cysteine residues as the final step in FK228 biosynthesis. Acquisition of the FK228 biosynthetic gene cluster and acclimation of an efficient genetic system should enable genetic engineering of the FK228 biosynthetic pathway in C. violaceum no. 968 for the generation of structural analogs as anticancer drug candidates. PMID- 17400766 TI - Variable-number tandem repeats as molecular markers for biotypes of Pasteuria ramosa in Daphnia spp. AB - Variable-number tandem repeats (VNTRs) have been identified in populations of Pasteuria ramosa, a castrating endobacterium of Daphnia species. The allelic polymorphisms at 14 loci in laboratory and geographically diverse soil samples showed that VNTRs may serve as biomarkers for the genetic characterization of P. ramosa isolates. PMID- 17400767 TI - PsrA, the Pseudomonas sigma regulator, controls regulators of epiphytic fitness, quorum-sensing signals, and plant interactions in Pseudomonas syringae pv. tomato strain DC3000. AB - Pseudomonas syringae pv. tomato strain DC3000, a pathogen of tomato and Arabidopsis, occurs as an epiphyte. It produces N-acyl homoserine lactones (AHLs) which apparently function as quorum-sensing signals. A Tn5 insertion mutant of DC3000, designated PsrA(-) (Psr is for Pseudomonas sigma regulator), overexpresses psyR (a LuxR-type regulator of psyI) and psyI (the gene for AHL synthase), and it produces a ca. 8-fold-higher level of AHL than does DC3000. The mutant is impaired in its ability to elicit the hypersensitive reaction and is attenuated in its virulence in tomato. These phenotypes correlate with reduced expression of hrpL, the gene for an alternate sigma factor, as well as several hrp and hop genes during early stages of incubation in a Hrp-inducing medium. PsrA also positively controls rpoS, the gene for an alternate sigma factor known to control various stress responses. By contrast, PsrA negatively regulates rsmA1, an RNA-binding protein gene known to function as negative regulator, and aefR, a tetR-like gene known to control AHL production and epiphytic fitness in P. syringae pv. syringae. Gel mobility shift assays and other lines of evidence demonstrate a direct interaction of PsrA protein with rpoS promoter DNA and aefR operator DNA. In addition, PsrA negatively autoregulates and binds the psrA operator. In an AefR(-) mutant, the expression of psyR and psyI and AHL production are lower than those in DC3000, the AefR(+) parent. In an RpoS(-) mutant, on the other hand, the levels of AHL and transcripts of psyR and psyI are much higher than those in the RpoS(+) parent, DC3000. We present evidence, albeit indirect, that the RpoS effect occurs via psyR. Thus, AefR positively regulates AHL production, whereas RpoS has a strong negative effect. We show that AefR and RpoS do not regulate PsrA and that the PsrA effect on AHL production is exerted via its cumulative, but independent, effects on both AefR and RpoS. PMID- 17400768 TI - Use of PCR-targeted mutagenesis to disrupt production of fusaricidin-type antifungal antibiotics in Paenibacillus polymyxa. AB - Paenibacillus polymyxa (formerly Bacillus polymyxa) PKB1 has been identified as a potential agent for biocontrol of blackleg disease of canola, caused by the pathogenic fungus Leptosphaeria maculans. The factors presumed to contribute to disease suppression by strain PKB1 include the production of fusaricidin-type antifungal metabolites that appear around the onset of bacterial sporulation. The fusaricidins are a family of lipopeptide antibiotics consisting of a beta-hydroxy fatty acid linked to a cyclic hexapeptide. Using a reverse genetic approach based on conserved motifs of nonribosomal peptide synthetases, a DNA fragment that appears to encode the first two modules of the putative fusaricidin synthetase (fusA) was isolated from PKB1. To confirm the involvement of fusA in production of fusaricidins, a modified PCR targeting mutagenesis protocol was developed to create a fusA mutation in PKB1. A DNA fragment internal to fusA was replaced by a gene disruption cassette containing two antibiotic resistance genes for independent selection of apramycin resistance in Escherichia coli and chloramphenicol resistance in P. polymyxa. Inclusion of an oriT site in the disruption cassette allowed efficient transfer of the inactivated fusA allele to P. polymyxa by intergeneric conjugation. Targeted disruption of fusA led to the complete loss of antifungal activity against L. maculans, suggesting that fusA plays an essential role in the nonribosomal synthesis of fusaricidins. PMID- 17400770 TI - Diversity and abundance of nitrate reductase genes (narG and napA), nitrite reductase genes (nirS and nrfA), and their transcripts in estuarine sediments. AB - Estuarine systems are the major conduits for the transfer of nitrate from agricultural and other terrestrial-anthropogenic sources into marine ecosystems. Within estuarine sediments some microbially driven processes (denitrification and anammox) result in the net removal of nitrogen from the environment, while others (dissimilatory nitrate reduction to ammonium) do not. In this study, molecular approaches have been used to investigate the diversity, abundance, and activity of the nitrate-reducing communities in sediments from the hypernutrified Colne estuary, United Kingdom, via analysis of nitrate and nitrite reductase genes and transcripts. Sequence analysis of cloned PCR-amplified narG, napA, and nrfA gene sequences showed the indigenous nitrate-reducing communities to be both phylogenetically diverse and also divergent from previously characterized nitrate reduction sequences in soils and offshore marine sediments and from cultured nitrate reducers. In both the narG and nrfA libraries, the majority of clones (48% and 50%, respectively) were related to corresponding sequences from delta proteobacteria. A suite of quantitative PCR primers and TaqMan probes was then developed to quantify phylotype-specific nitrate (narG and napA) and nitrite reductase (nirS and nrfA) gene and transcript numbers in sediments from three sites along the estuarine nitrate gradient. In general, both nitrate and nitrite reductase gene copy numbers were found to decline significantly (P < 0.05) from the estuary head towards the estuary mouth. The development and application, for the first time, of quantitative reverse transcription-PCR assays to quantify mRNA sequences in sediments revealed that transcript numbers for three of the five phylotypes quantified were greatest at the estuary head. PMID- 17400771 TI - Sensitive whole-cell biosensor suitable for detecting a variety of N-acyl homoserine lactones in intact rhizosphere microbial communities. AB - To investigate quorum sensing in rhizosphere soil, a whole-cell biosensor, Agrobacterium tumefaciens(pAHL-Ice), was constructed. The biosensor responded to all N-acyl homoserine lactones (AHLs) tested, except C(4) homoserine lactone, with a minimum detection limit of 10(-12) M, as well as to both exogenously added AHLs and AHL-producing bacterial strains in soil. This highly sensitive biosensor reveals for the first time the increased AHL availability in intact rhizosphere microbial communities compared to that in bulk soil. PMID- 17400769 TI - Geographical distribution and diversity of bacteria associated with natural populations of Drosophila melanogaster. AB - Drosophila melanogaster is one of the most widely used model systems in biology. However, little is known about its associated bacterial community. As a first step towards understanding these communities, we compared bacterial 16S rRNA gene sequence libraries recovered from 11 natural populations of adult D. melanogaster. Bacteria from these sequence libraries were grouped into 74 distinct taxa, spanning the phyla Proteobacteria, Bacteroidetes, and Firmicutes, which were unevenly spread across host populations. Summed across populations, the distribution of abundance of genera was closely fit by a power law. We observed differences among host population locations both in bacterial community richness and in composition. Despite this significant spatial variation, no relationship was observed between species richness and a variety of abiotic factors, such as temperature and latitude. Overall, bacterial communities associated with adult D. melanogaster hosts are diverse and differ across host populations. PMID- 17400772 TI - Seasonal variations in the contributions of different bacterial groups to the uptake of low-molecular-weight compounds in northwestern Mediterranean coastal waters. AB - We analyzed the contributions of different heterotrophic bacterial groups to the uptake of several low-molecular weight compounds during a seasonal cycle on the northwestern Mediterranean coast (Blanes Bay Microbial Observatory). The bacterial assemblage structure had been shown to change substantially year-round for this site, but whether changes in the activities of the different bacterial groups also occurred on the seasonal scale was unknown. Microautoradiography combined with catalyzed reporter deposition fluorescence in situ hybridization was used to analyze the patterns of glucose, amino acid, and ATP uptake by different bacterial groups. Gammaproteobacteria and Bacteroidetes were not very active in the uptake of glucose at any time of the year (<10% of cells were active) compared to Alphaproteobacteria (generally >20% of cells were active). Dissolved free amino acids were taken up considerably by Alphaproteobacteria and Gammaproteobacteria but not by Bacteroidetes. Relatively high percentages of cells of the three broad phylogenetic groups actively took up ATP, which could be related to the important phosphorous limitation of bacterial production during most of the year in Blanes Bay. The contribution of SAR11 to the uptake of the monomers was variable year-round, generally with fewer than 30% of the cells being active. By contrast, Roseobacter were highly overrepresented in the uptake of all the substrates throughout all the year, with more than 50% of cells being active in all the samples and for all substrates. Our results suggest that substantial changes in the activity of some phylogenetic groups of bacteria occur throughout the year. PMID- 17400773 TI - Development of high-throughput phenotyping of metagenomic clones from the human gut microbiome for modulation of eukaryotic cell growth. AB - Metagenomic libraries derived from human intestinal microbiota (20,725 clones) were screened for epithelial cell growth modulation. Modulatory clones belonging to the four phyla represented among the metagenomic libraries were identified (hit rate, 0.04 to 8.7% depending on the screening cutoff). Several candidate loci were identified by transposon mutagenesis and subcloning. PMID- 17400774 TI - A marine Mesorhizobium sp. produces structurally novel long-chain N-acyl-L homoserine lactones. AB - Our study focused on a Mesorhizobium sp. that is phylogenetically affiliated by 16S rRNA gene sequence to other marine and saline bacteria of this genus. Liquid chromatography-mass spectrometry investigations of the extract obtained from solid-phase extraction of cultures of this bacterium indicated the presence of several N-acyl homoserine lactones (AHLs), with chain lengths of C(10) to C(16). Chromatographic separation of the active bacterial extract yielded extraordinarily large amounts of two unprecedented acylated homoserine lactones, 5-cis-3-oxo-C(12)-homoserine lactone (5-cis-3-oxo-C(12)-HSL) (compound 1) and 5 cis-C(12)-HSL (compound 2). Quorum-sensing activity of compounds 1 and 2 was shown in two different biosensor systems [Escherichia coli MT102(pSB403) and Pseudomonas putida F117(pKR-C12)]. Furthermore, it was shown that both compounds can restore protease and pyoverdin production of an AHL-deficient Pseudomonas aeruginosa PAO1 lasI rhlI double mutant, suggesting that these signal molecules maybe used for intergenus signaling. In conclusion, these data indicate that the quorum-sensing activity of compounds 1 and 2 is modulated by the chain length and functional groups of the acyl moiety. Additionally, compound 1 showed antibacterial and cytotoxic activities. PMID- 17400775 TI - Molecular characterization of a glucokinase with broad hexose specificity from Bacillus sphaericus strain C3-41. AB - Bacillus sphaericus cannot metabolize sugar since it lacks several of the enzymes necessary for glycolysis. Our results confirmed the presence of a glucokinase encoding gene, glcK, and a phosphofructokinase-encoding gene, pfk, on the bacterial chromosome and expression of glucokinase during vegetative growth of B. sphaericus strains. However, no phosphoglucose isomerase gene (pgi) or phosphoglucose isomerase enzyme activity was detected in these strains. Furthermore, one glcK open reading frame was cloned from B. sphaericus strain C3 41 and then expressed in Escherichia coli. Biochemical analysis revealed that this gene encoded a protein with a molecular mass of 33 kDa and that the purified recombinant glucokinase had K(m) values of 0.52 and 0.31 mM for ATP and glucose, respectively. It has been proved that this ATP-dependent glucokinase can also phosphorylate fructose and mannose, and sequence alignment of the glcK gene indicated that it belongs to the ROK protein family. It is postulated that the absence of the phosphoglucose isomerase-encoding gene pgi in B. sphaericus might be one of the reasons for the inability of this bacterium to metabolize carbohydrates. Our findings provide additional data that further elucidate the specific metabolic pathway and could be used for genetic improvement of B. sphaericus. PMID- 17400776 TI - Genome sequence of the cellulolytic gliding bacterium Cytophaga hutchinsonii. AB - The complete DNA sequence of the aerobic cellulolytic soil bacterium Cytophaga hutchinsonii, which belongs to the phylum Bacteroidetes, is presented. The genome consists of a single, circular, 4.43-Mb chromosome containing 3,790 open reading frames, 1,986 of which have been assigned a tentative function. Two of the most striking characteristics of C. hutchinsonii are its rapid gliding motility over surfaces and its contact-dependent digestion of crystalline cellulose. The mechanism of C. hutchinsonii motility is not known, but its genome contains homologs for each of the gld genes that are required for gliding of the distantly related bacteroidete Flavobacterium johnsoniae. Cytophaga-Flavobacterium gliding appears to be novel and does not involve well-studied motility organelles such as flagella or type IV pili. Many genes thought to encode proteins involved in cellulose utilization were identified. These include candidate endo-beta-1,4 glucanases and beta-glucosidases. Surprisingly, obvious homologs of known cellobiohydrolases were not detected. Since such enzymes are needed for efficient cellulose digestion by well-studied cellulolytic bacteria, C. hutchinsonii either has novel cellobiohydrolases or has an unusual method of cellulose utilization. Genes encoding proteins with cohesin domains, which are characteristic of cellulosomes, were absent, but many proteins predicted to be involved in polysaccharide utilization had putative D5 domains, which are thought to be involved in anchoring proteins to the cell surface. PMID- 17400777 TI - Biochemical and ecological control of geosmin and 2-methylisoborneol in source waters. PMID- 17400778 TI - Cloning, functional characterization, and mode of action of a novel insecticidal pore-forming toxin, sphaericolysin, produced by Bacillus sphaericus. AB - An insecticidal protein produced by Bacillus sphaericus A3-2 was purified to elucidate its structure and mode of action. The active principle purified from the culture broth of A3-2 was a protein with a molecular mass of 53 kDa that rapidly intoxicated German cockroaches (Blattela germanica) at a dose of about 100 ng when injected. The insecticidal protein sphaericolysin possessed the undecapeptide motif of cholesterol-dependent cytolysins and had a unique N terminal sequence. The recombinant protein expressed in Escherichia coli was equally as potent as the native protein. Sphaericolysin-induced hemolysis resulted from the protein's pore-forming action. This activity as well as the insecticidal activity was markedly reduced by a Y159A mutation. Also, coapplication of sphaericolysin with cholesterol abolished the insecticidal action, suggesting that cholesterol binding plays an important role in insecticidal activity. Sphaericolysin-lysed neurons dissociated from the thoracic ganglia of the German cockroaches. In addition, sphaericolysin's activity in ganglia was suppressed by the Y159A mutation. The sphaericolysin-induced damage to the cockroach ganglia was greater than the damage to the ganglia of common cutworms (Spodoptera litura), which accounts, at least in part, for the higher sensitivity to sphaericolysin displayed by the cockroaches than that displayed by cutworms. PMID- 17400779 TI - Responses of rhizobia to desiccation in relation to osmotic stress, oxygen, and temperature. PMID- 17400780 TI - Identification of unconventional intestinal pathogenic Escherichia coli isolates expressing intermediate virulence factor profiles by using a novel single-step multiplex PCR. AB - Intestinal pathogenic Escherichia coli represents a global health problem for mammals, including humans. At present, diarrheagenic E. coli bacteria are grouped into seven major pathotypes that differ in their virulence factor profiles, severity of clinical manifestations, and prognosis. In this study, we developed and evaluated a one-step multiplex PCR (MPCR) for the straightforward differential identification of intestinal pathotypes of E. coli. The specificity of this novel MPCR was validated by using a subset of reference strains and further confirmed by PCR-independent pheno- and genotypic characterization. Moreover, we tested 246 clinical E. coli isolates derived from diarrhea patients from several distinct geographic regions. Interestingly, besides strains belonging to the defined and well-described pathotypes, we identified five unconventional strains expressing intermediate virulence factor profiles. These strains have been further characterized and appear to represent intermediate strains carrying genes and expressing factors associated with enteropathogenic E. coli, Shiga toxin-producing E. coli, enterotoxigenic E. coli, and enteroaggregative E. coli alike. These strains represent further examples of the extraordinary plasticity of the E. coli genome. Moreover, this implies that the important identification of specific pathotypes has to be based on a broad matrix of indicator genes. In addition, the presence of intermediate strains needs to be accounted for. PMID- 17400781 TI - Differentiation of Bacillus anthracis, B. cereus, and B. thuringiensis by using pulsed-field gel electrophoresis. AB - A pulsed-field gel electrophoresis (PFGE) method was developed for discriminating Bacillus anthracis from B. cereus and B. thuringiensis. A worldwide collection of 25 B. anthracis isolates showed high-profile homology, and these isolates were unambiguously distinguished from B. cereus and B. thuringiensis isolates by cluster analysis of the whole-genome macrorestriction enzyme digestion patterns generated by NotI. PMID- 17400782 TI - Monochloramine inactivation of bacterial select agents. AB - Seven species of bacterial select agents were tested for susceptibility to monochloramine. Under test conditions, the monochloramine routinely maintained in potable water would reduce six of the species by 2 orders of magnitude within 4.2 h. Bacillus anthracis spores would require up to 3.5 days for the same inactivation with monochloramine. PMID- 17400783 TI - A homologue of the Aspergillus velvet gene regulates both cephalosporin C biosynthesis and hyphal fragmentation in Acremonium chrysogenum. AB - The Aspergillus nidulans velvet (veA) gene encodes a global regulator of gene expression controlling sexual development as well as secondary metabolism. We have identified the veA homologue AcveA from Acremonium chrysogenum, the major producer of the beta-lactam antibiotic cephalosporin C. Two different disruption strains as well as the corresponding complements were generated as a prelude to detailed functional analysis. Northern hybridization and quantitative real-time PCR clearly indicate that the nucleus-localized AcVEA polypeptide controls the transcriptional expression of six cephalosporin C biosynthesis genes. The most drastic reduction in expression is seen for cefEF, encoding the deacetoxycephalosporine/deacetylcephalosporine synthetase. After 120 h of growth, the cefEF transcript level is below 15% in both disruption strains compared to the wild type. These transcriptional expression data are consistent with results from a comparative and time-dependent high-performance liquid chromatography analysis of cephalosporin C production. Compared to the recipient, both disruption strains have a cephalosporin C titer that is reduced by 80%. In addition to its role in cephalosporin C biosynthesis, AcveA is involved in the developmentally dependent hyphal fragmentation. In both disruption strains, hyphal fragmentation is already observed after 48 h of growth, whereas in the recipient strain, arthrospores are not even detected before 96 h of growth. Finally, the two mutant strains show hyperbranching of hyphal tips on osmotically nonstabilized media. Our findings will be significant for biotechnical processes that require a defined stage of cellular differentiation for optimal production of secondary metabolites. PMID- 17400784 TI - Shiga toxin gene loss and transfer in vitro and in vivo during enterohemorrhagic Escherichia coli O26 infection in humans. AB - Escherichia coli serogroup O26 consists of enterohemorrhagic E. coli (EHEC) and atypical enteropathogenic E. coli (aEPEC). The former produces Shiga toxins (Stx), major determinants of EHEC pathogenicity, encoded by bacteriophages; the latter is Stx negative. We have isolated EHEC O26 from patient stools early in illness and aEPEC O26 from stools later in illness, and vice versa. Intrapatient EHEC and aEPEC isolates had quite similar pulsed-field gel electrophoresis (PFGE) patterns, suggesting that they might have arisen by conversion between the EHEC and aEPEC pathotypes during infection. To test this hypothesis, we asked whether EHEC O26 can lose stx genes and whether aEPEC O26 can be lysogenized with Stx encoding phages from EHEC O26 in vitro. The stx2 loss associated with the loss of Stx2-encoding phages occurred in 10% to 14% of colonies tested. Conversely, Stx2- and, to a lesser extent, Stx1-encoding bacteriophages from EHEC O26 lysogenized aEPEC O26 isolates, converting them to EHEC strains. In the lysogens and EHEC O26 donors, Stx2-converting bacteriophages integrated in yecE or wrbA. The loss and gain of Stx-converting bacteriophages diversifies PFGE patterns; this parallels findings of similar but not identical PFGE patterns in the intrapatient EHEC and aEPEC O26 isolates. EHEC O26 and aEPEC O26 thus exist as a dynamic system whose members undergo ephemeral interconversions via loss and gain of Stx-encoding phages to yield different pathotypes. The suggested occurrence of this process in the human intestine has diagnostic, clinical, epidemiological, and evolutionary implications. PMID- 17400785 TI - High-resolution DNA melt curve analysis of the clustered, regularly interspaced short-palindromic-repeat locus of Campylobacter jejuni. AB - A novel method for genotyping the clustered, regularly interspaced short palindromic-repeat (CRISPR) locus of Campylobacter jejuni is described. Following real-time PCR, CRISPR products were subjected to high-resolution melt (HRM) analysis, a new technology that allows precise melt profile determination of amplicons. This investigation shows that the CRISPR HRM assay provides a powerful addition to existing C. jejuni genotyping methods and emphasizes the potential of HRM for genotyping short sequence repeats in other species. PMID- 17400786 TI - Diversification of lupine Bradyrhizobium strains: evidence from nodulation gene trees. AB - Bradyrhizobium strains isolated in Europe from Genisteae and serradella legumes form a distinct lineage, designated clade II, on nodulation gene trees. Clade II bradyrhizobia appear to prevail also in the soils of Western Australia and South Africa following probably accidental introduction with seeds of their lupine and serradella hosts. Given this potential for dispersal, we investigated Bradyrhizobium isolates originating from a range of native New World lupines, based on phylogenetic analyses of nodulation (nodA, nodZ, noeI) and housekeeping (atpD, dnaK, glnII, recA) genes. The housekeeping gene trees revealed considerable diversity among lupine bradyrhizobia, with most isolates placed in the Bradyrhizobium japonicum lineage, while some European strains were closely related to Bradyrhizobium canariense. The nodA gene tree resolved seven strongly supported groups (clades I to VII) that correlated with strain geographical origins and to some extent with major Lupinus clades. All European strains were placed in clade II, whereas only a minority of New World strains was placed in this clade. This work, as well as our previous studies, suggests that clade II diversified predominately in the Old World, possibly in the Mediterranean. Most New World isolates formed subclade III.2, nested in a large "pantropical" clade III, which appears to be New World in origin, although it also includes strains originating from nonlupine legumes. Trees generated using nodZ and noeI gene sequences accorded well with the nodA tree, but evidence is presented that the noeI gene may not be required for nodulation of lupine and that loss of this gene is occurring. PMID- 17400787 TI - Identification of novel genes involved in long-chain n-alkane degradation by Acinetobacter sp. strain DSM 17874. AB - Acinetobacter sp. strain DSM 17874 is capable of utilizing n-alkanes with chain lengths ranging from that of decane (C10H22) to that of tetracontane (C40H82) as a sole carbon source. Two genes encoding AlkB-type alkane hydroxylase homologues, designated alkMa and alkMb, have been shown to be involved in the degradation of n-alkanes with chain lengths of from 10 to 20 C atoms in this strain. Here, we describe a novel high-throughput screening method and the screening of a transposon mutant library to identify genes involved in the degradation of n alkanes with C chain lengths longer than 20, which are solid at 30 degrees C, the optimal growth temperature for Acinetobacter sp. strain DSM 17874. A library consisting of approximately 6,800 Acinetobacter sp. strain DSM 17874 transposon mutants was constructed and screened for mutants unable to grow on dotriacontane (C32H66) while simultaneously showing wild-type growth characteristics on shorter chain n-alkanes. For 23 such mutants isolated, the genes inactivated by transposon insertion were identified. Targeted inactivation and complementation studies of one of these genes, designated almA and encoding a putative flavin binding monooxygenase, confirmed its involvement in the strain's metabolism of long-chain n-alkanes. To our knowledge, almA represents the first cloned gene shown to be involved in the bacterial degradation of long-chain n-alkanes of 32 C's and longer. Genes encoding AlmA homologues were also identified in other long chain n-alkane-degrading Acinetobacter strains. PMID- 17400788 TI - Coccoid form of Helicobacter pylori as a morphological manifestation of cell adaptation to the environment. AB - After characterization of preferred conditions for Helicobacter pylori survival in the sessile state, it was observed that the bacterium transforms from spiral to coccoid under mild circumstances, whereas under extreme ones it is unable to undergo shape modification. This strongly supports the view that transformation into the coccoid form is an active, biologically led process, switched on by the bacterium as a protection mechanism. PMID- 17400789 TI - Phylogenetic diversity of gram-positive bacteria cultured from marine sediments. AB - Major advances in our understanding of marine bacterial diversity have been gained through studies of bacterioplankton, the vast majority of which appear to be gram negative. Less effort has been devoted to studies of bacteria inhabiting marine sediments, yet there is evidence to suggest that gram-positive bacteria comprise a relatively large proportion of these communities. To further expand our understanding of the aerobic gram-positive bacteria present in tropical marine sediments, a culture-dependent approach was applied to sediments collected in the Republic of Palau from the intertidal zone to depths of 500 m. This investigation resulted in the isolation of 1,624 diverse gram-positive bacteria spanning 22 families, including many that appear to represent new taxa. Phylogenetic analysis of 189 representative isolates, based on 16S rRNA gene sequence data, indicated that 124 (65.6%) belonged to the class Actinobacteria while the remaining 65 (34.4%) were members of the class Bacilli. Using a sequence identity value of >/=98%, the 189 isolates grouped into 78 operational taxonomic units, of which 29 (37.2%) are likely to represent new taxa. The high degree of phylogenetic novelty observed during this study highlights the fact that a great deal remains to be learned about the diversity of gram-positive bacteria in marine sediments. PMID- 17400790 TI - Autism, language delay and mental retardation in a patient with 7q11 duplication. AB - BACKGROUND: Chromosomal rearrangements, arising from unequal recombination between repeated sequences, are found in a subset of patients with autism. Duplications involving loci associated with behavioural disturbances constitute an especially good candidate mechanism. The Williams-Beuren critical region (WBCR), located at 7q11.23, is commonly deleted in Williams-Beuren microdeletion syndrome (WBS). However, only four patients with a duplication of the WBCR have been reported to date: one with severe language delay and the three others with variable developmental, psychomotor and language delay. OBJECTIVE AND METHODS: In this study, we screened 206 patients with autism spectrum disorders for the WBCR duplication by quantitative microsatellite analysis and multiple ligation dependent probe amplification. RESULTS: We identified one male patient with a de novo interstitial duplication of the entire WBCR of paternal origin. The patient had autistic disorder, severe language delay and mental retardation, with very mild dysmorphic features. CONCLUSION: We report the first patient with autistic disorder and a WBCR duplication. This observation indicates that the 7q11.23 duplication could be involved in complex clinical phenotypes, ranging from developmental or language delay to mental retardation and autism, and extends the phenotype initially reported. These findings also support the existence of one or several genes in 7q11.23 sensitive to gene dosage and involved in the development of language and social interaction. PMID- 17400791 TI - A survey of locus-specific database curation. Human Genome Variation Society. AB - It is widely accepted that curation of variation in genes is best performed by experts in those genes and their variation. However, obtaining funding for such variation is difficult even though up-to-date lists of variations in genes are essential for optimum delivery of genetic healthcare and for medical research. This study was undertaken to gather information on gene-specific databases (locus specific databases) in an effort to understand their functioning, funding and needs. A questionnaire was sent to 125 curators and we received 47 responses. Individuals performed curation of up to 69 genes. The time curators spent curating was extremely variable. This ranged from 0 h per week up to 5 curators spending over 4 h per week. The funding required ranged from US$600 to US$45,000 per year. Most databases were stimulated by the Human Genome Organization Mutation Database Initiative and used their guidelines. Many databases reported unpublished mutations, with all but one respondent reporting errors in the literature. Of the 13 who reported hit rates, 9 reported over 52,000 hits per year. On the basis of this, five recommendations were made to improve the curation of variation information, particularly that of mutations causing single gene disorder: 1. A curator for each gene, who is an expert in it, should be identified or nominated. 2. Curation at a minimum of 2 h per week at US$2000 per gene per year should be encouraged. 3. Guidelines and custom software use should be encouraged to facilitate easy setup and curation. 4. Hits per week on the website should be recorded to allow the importance of the site to be illustrated for grant-giving purposes. 5. Published protocols should be followed in the establishment of locus-specific databases. PMID- 17400792 TI - The Shwachman-Bodian-Diamond syndrome gene mutations cause a neonatal form of spondylometaphysial dysplasia (SMD) resembling SMD Sedaghatian type. AB - The Shwachman-Bodian-Diamond syndrome (SBDS) gene is a causative gene for Shwachman-Diamond syndrome, an autosomal recessive disorder with exocrine pancreatic insufficiency, bone marrow dysfunction and skeletal dysplasia. We report here on two patients with skeletal manifestations at the severest end of the phenotypic spectrum of SBDS mutations. An 11-year-old Japanese girl presented with neonatal respiratory failure necessitating lifelong ventilation support, severe short stature and severe developmental delay. She developed neutropenia in infancy, and decreased serum amylase was noted in childhood. A British boy was a stillbirth with pulmonary hypoplasia and hepatic fibrosis found on autopsy. Both cases had neonatal skeletal manifestations that included platyspondyly, lacy iliac crests and severe metaphysial dysplasia, and thus did not fall in the range of the known Shwachman-Diamond syndrome skeletal phenotype but resembled spondylometaphysial dysplasia (SMD) Sedaghatian type. The girl harboured a recurrent mutation (183TA-->CT) and a novel missense mutation (79T-->C), whereas the boy carried two recurrent mutations (183TA-->CT and 258+2T-->C). We also examined SBDS in one typical case with SMD Sedaghantian type and eight additional cases with neonatal SMD, but failed to discover SBDS mutations. Our experience expands the phenotypic spectrum of SBDS mutations, which, at its severest end, results in severe neonatal SMD. PMID- 17400793 TI - Mutations in the ND5 subunit of complex I of the mitochondrial DNA are a frequent cause of oxidative phosphorylation disease. AB - BACKGROUND: Detection of mutations in the mitochondrial DNA (mtDNA) is usually limited to common mutations and the transfer RNA genes. However, mutations in other mtDNA regions can be an important cause of oxidative phosphorylation (OXPHOS) disease as well. OBJECTIVE: To investigate whether regions in the mtDNA are preferentially mutated in patients with OXPHOS disease. METHODS: Screening of the mtDNA for heteroplasmic mutations was performed by denaturing high performance liquid chromatography analysis of 116 patients with OXPHOS disease but without the common mtDNA mutations. RESULTS: An mtDNA sequence variant was detected in 15 patients, 5 of which were present in the ND5 gene. One sequence variant was new and three were known, one of which was found twice. The novel sequence variant m.13511A-->T occurred in a patient with a Leigh-like syndrome. The known mutation m.13513G-->A, associated with mitochondrial encephalomyopathy lactic acidosis and stroke-like syndrome (MELAS) and MELAS/Leigh/Leber hereditary optic neuropathy overlap syndrome, was found in a relatively low percentage in two patients from two different families, one with a MELAS/Leigh phenotype and one with a MELAS/chronic progressive external ophthalmoplegia phenotype. The known mutation m.13042G-->A, detected previously in a patient with a MELAS/myoclonic epilepsy, ragged red fibres phenotype and in a family with a prevalent ocular phenotype, was now found in a patient with a Leigh-like phenotype. The sequence variant m.12622G-->A was reported once in a control database as a polymorphism, but is reported in this paper as heteroplasmic in three brothers, all with infantile encephalopathy (Leigh syndrome) fatal within the first 15 days of life. Therefore, a causal relationship between the presence of this sequence variant and the onset of mitochondrial disease cannot be entirely excluded at this moment. CONCLUSIONS: Mutation screening of the ND5 gene is advised for routine diagnostics of patients with OXPHOS disease, especially for those with MELAS- and Leigh-like syndrome with a complex I deficiency. PMID- 17400794 TI - Array comparative genomic hybridisation analysis of boys with X-linked hypopituitarism identifies a 3.9 Mb duplicated critical region at Xq27 containing SOX3. PMID- 17400795 TI - Phenocopies in breast cancer 1 (BRCA1) families: implications for genetic counselling. PMID- 17400796 TI - Hypomethylation in the 11p15 telomeric imprinting domain in a patient with Silver Russell syndrome with a CSH1 deletion (17q24) renders a functional role of this alteration unlikely. PMID- 17400797 TI - The leptin-dependent and -independent melanocortin signaling system: regulation of feeding and energy expenditure. AB - The brain hypothalamus coordinates extra-hypothalamic regions to maintain energy homeostasis through the regulation of food intake and energy expenditure. A number of anorexigenic and orexigenic molecules in the hypothalamic nuclei participate in the control of energy homeostasis. Leptin and pro-opiomelanocortin (POMC)-derived alpha-melanocyte-stimulating hormone are key anorectic molecules, and the leptin receptor and POMC gene are both expressed in the hypothalamic arcuate nucleus. Although it has been considered that melanocortin signaling is localized downstream to leptin signaling, data have accumulated to support the concept of a leptin-independent melanocortin signaling system. We focus on and review the melanocortin signaling system that functions dependently or independently of leptin signaling in the regulation of energy homeostasis. PMID- 17400798 TI - Effects of the CRF1 receptor antagonist SSR125543 on energy balance and food deprivation-induced neuronal activation in obese Zucker rats. AB - The corticotropin-releasing factor (CRF) system is involved in numerous physiological and behavioral actions, including the regulation of energy balance. We examined the effects of the CRF(1) receptor antagonist, SSR125543, on energy balance and food deprivation-induced neuronal activation in obese rats. Lean (Fa/?) and obese (fa/fa) Zucker rats were treated orally with SSR125543 at a daily dose of 30 mg/kg for 21 days. Rats were killed either fed ad libitum or food deprived for 6 h in order to induce a mild stress response in obese rats. SSR125543 reduced plasma corticosterone levels in lean rats, prevented corticosterone response to fasting in obese rats, and increased CRF mRNA levels in the paraventricular hypothalamic nucleus (PVN) of both lean and obese rats, further confirming that the antagonist partially blocked CRF(1) receptors. SSR125543 increased protein gain in obese rats. Whole carcass analyses showed reduced energy and fat gains in lean rats. Consistent with reduced fat gain, circulating triglyceride and leptin levels were reduced in SSR125543-treated lean rats. In obese rats, circulating glucose levels and the homeostasis model assessment of insulin resistance index of insulin resistance were reduced by SSR125543 treatment. CRF(1) receptor blockade increased uncoupling protein-1 mRNA levels in interscapular brown adipose tissue of obese rats. The antagonist partly blocked the fasting-induced changes in c-fos mRNA levels in the PVN and arcuate nucleus of obese rats. Overall, these results suggest that although SSR125543 had relatively mild effects on energy balance, CRF(1) receptor blockade attenuated several metabolic effects of short-term fasting and improved plasma variables related to the metabolic syndrome and diabetes. PMID- 17400799 TI - Thyroid hormone receptor beta-specific agonist GC-1 increases energy expenditure and prevents fat-mass accumulation in rats. AB - It is well known that thyroid hormone affects body composition; however, the effect of the thyroid hormone receptor beta (TRbeta)-selective thyromimetic GC-1 on this biological feature had not been demonstrated. In the current study, we compared the effects of a 6-week treatment with triiodothyronine (T3; daily injections of 3 or 6 microg/100 g body weight) or GC-1 (equimolar doses) on different metabolic parameters in adult female rats. Whereas all animals gained weight (17-25 g) in a way not basically affected by T3 or GC-1 treatment, only T3 treatment selectively increased food intake (50-70%). Oxygen consumption was significantly and equally increased (50-70%) by T3 and GC-1. Analysis of body composition by dual-energy X-ray absorptiometry (DEXA) revealed that, whereas control animals gained about 80% of fat mass, T3- or GC-1-treated animals lost 70 90 and approximately 20% respectively. Direct analysis of the carcass showed that T3 treatment promoted a 14-74% decrease in fat content but GC-1 treatment promoted only a 15-23% reduction. The gain in lean mass by DEXA and the carcass protein content were not affected by T3 or GC-1 treatment. However, the mass of individual skeletal muscles was negatively affected by T3 but only barely by GC 1. These findings highlight the potential use of GC-1 for the treatment of obesity and the metabolic syndrome. PMID- 17400800 TI - Prenatal influences on susceptibility to diet-induced obesity are mediated by altered neuroendocrine gene expression. AB - The escalating rates of obesity and type 2 diabetes have reached pandemic proportions. It has been proposed that the risk of developing metabolic disorders in adult life is influenced by environmental factors, which operate during the early periods of development. We have previously shown that an interaction between the prenatal and the postnatal dietary environment amplifies the propensity towards diet-induced obesity, although the mechanisms are unclear. In the present study, we investigated the interaction between prenatal undernutrition and postnatal high-fat nutrition on key genes of the hypothalamic appetite regulatory network. Pregnant Wistar rats were fed a standard chow diet either ad libitum (AD) or at 30% of AD intake throughout gestation (UN). From weaning, female AD and UN offspring were fed either a standard chow (ADC n = 8, UNC n = 8) or a high-fat diet (45% kcal as fat; ADHF n = 8, UNHF n = 8) ad libitum for the remainder of the study. At 24 weeks of age, body composition was assessed by dual energy X-ray absorptiometry analysis and total RNA was extracted from whole rat hypothalami. Real-time PCR was performed to characterise pro opiomelanocortin (POMC), neuropeptide Y (NPY), agouti-related protein (AgRP) and OBRb gene expression at the mRNA level. Our results demonstrate that the amplification of postnatal obesity develops as a consequence of an interaction between prenatal under-nutrition and postnatal high-fat nutrition. This phenotype also shows significant alterations in POMC, NPY, AgRP and OBRb gene expression together with elevations in circulating levels of both plasma leptin and insulin. These findings are consistent with the predictive adaptive response hypothesis that neuroendocrine development during fetal life may be based on predictions about postnatal environmental conditions. Increased susceptibility to diet induced obesity develops if a mismatch between the anticipated and the actual conditions are encountered. PMID- 17400801 TI - Peroxisome proliferator-activated receptor alpha deficiency modifies glucose handling by isolated mouse adipocytes. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a transcription factor that regulates enzymes involved in fatty acid (FA) utilisation. PPARalpha null mice have recently been demonstrated to have increased whole-body glucose turnover in vivo. This has been attributed to increased glucose uptake by adipose tissue, but the impact of PPARalpha deficiency on the characteristics of glucose handling by isolated adipocytes ex vivo is unknown. To determine directly the impact of PPARalpha deficiency on adipocyte glucose handling, thereby excluding any influence of humoral/neuronal factors, we examined total glucose metabolism as well as glucose disposition towards alternative fates in epididymal adipocytes isolated from wild-type and PPARalphanull mice. Total glucose metabolism (oxidation, incorporation into FA and glycerol moieties of triglyceride (TAG) and conversion to lactate) was measured under basal conditions (low glucose) and 'stimulated lipogenic' conditions (high glucose + insulin). Adipocytes from PPARalpha null mice had higher rates of glucose metabolism under both basal and stimulated lipogenic conditions, with increased glucose utilisation both for oxidation and entry into the synthesis of the FA and glycerol components of lipid. In particular, the capacity of adipocytes from PPARalpha-deficient mice to utilise glucose for synthesis of the glycerol backbone of TAG was greatly enhanced under stimulated (high glucose + insulin) conditions. The increased use of glucose for the glycerol moiety of adipocyte TAG may therefore contribute to, and provide explanation for, enhanced glucose turnover in PPARalpha null mice. PMID- 17400802 TI - Calpain inhibition impairs glycogen syntheses in HepG2 hepatoma cells without altering insulin signaling. AB - Calpains are a family of non-lysosomal cytoplasmatic cysteine proteases. Since calpain 10 (CAPN10), a member of the calpain family of proteases, has been found to represent a putative diabetes susceptibility gene, it was argued that calpains may be involved in the development of type 2 diabetes. The functional role of calpains in insulin signaling and/or insulin action is, however, not clear. We investigated the effects of the calpains 1 and 2 inhibitor PD151746 on insulin signaling and insulin action in human hepatoma G2 cells (HepG2). HepG2 cells were incubated without (-PD) or with (+PD) 5.33 micromol/l PD151746 for different times and then stimulated with 100 nmol/l insulin for 0 (t(0)), 5 (t(5)), 15 (t(15)), 30 (t(30)), 45 (t(45)), and 60 (t(60)) min. After solubilization of the cells, insulin receptor kinase activity, tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1), IRS-1-associated phosphatidylinositol-3 kinase (PI3 kinase), PI3-kinase activity, Thr(308) phosphorlyation of Akt, amount of protein tyrosine phosphatase-epsilon (PTPepsilon), and glycogen synthase activity were determined. Incubation with PD151746 resulted in a significant reduction of insulin-stimulated glycogen synthesis compared with cells not pre-incubated with the calpain inhibitor (-PD: t(0), 4.90 +/- 1.20%; t(5), 5.90 +/- 1.02%; t(15), 5.29 +/- 0.95%; t(30), 5.60 +/- 1.10%; t(45), 5.52 +/- 0.90%; t(60), 5.67 +/- 0.97%;+PD: t(0), 4.56 +/- 1.10%; t(5), 6.16 +/- 1.05%; t(15), 7.52 +/- 1.09%; t(30), 7.68 +/- 1.10%; t(45), 8.28 +/- 0.89%; t(60), 7.69 +/- 0.98%; P < 0.05). Incubation with PD151746 significantly increased the protein amount of PTPepsilon in the cells after 12 h (-PD: t(1), 0.85 +/- 0.18 RU (Relative unit); t(8), 0.87 +/- 0.18 RU; t(12), 0.9 +/- 0.13 RU; +PD: t(1), 0.92 +/- 0.21 RU; t(8), 1.1 +/- 0.15 RU; t(12), 1.34 +/- 0.16 RU; P < 0.05). Calpain inhibition with PD151746 had no effect on the insulin stimulation of the investigated insulin signaling parameters. These results in HepG2 cells suggest that calpains play a role in the hepatic regulation of insulin-stimulated glycogen synthesis independent of the PI3-kinase/Akt signaling pathway. PMID- 17400803 TI - Regulation of Leydig cell steroidogenesis by extracellular signal-regulated kinase 1/2: role of protein kinase A and protein kinase C signaling. AB - The steroidogenic acute regulatory (StAR) protein plays a central role in the regulation of steroid biosynthesis. While steroidogenesis is influenced by many processes, their modes of actions, in a few cases, remain obscure. In this study, we explored the mechanism of action of one such signaling pathway, the extracellular signal-regulated kinase 1/2 (ERK1/2), in regulating StAR expression and steroidogenesis in conjunction with the protein kinase A (PKA) and protein kinase C (PKC) pathways. Using MA-10 mouse Leydig tumor cells, we demonstrate that the activation of PKC and PKA signaling, by phorbol-12-myristate-13-acetate (PMA) and dibutyryl cAMP (dbcAMP)/human chorionic gonadotropin (hCG) respectively, was able to phosphorylate ERK1/2, an event markedly decreased by an upstream kinase inhibitor, U0126. Treatment with PMA enhanced StAR protein expression (associated with a slight increase in progesterone synthesis) but not its phosphorylation (P-StAR), which, in contrast, coordinately increased in response to dbcAMP/hCG. Inhibition of ERK1/2 activity by U0126 decreased PMA treated StAR expression but increased dbcAMP/hCG-mediated StAR and P-StAR; however, progesterone levels were attenuated. U0126 was found to affect StAR expression and steroidogenesis both at the transcriptional and translational levels. Further studies demonstrated that the effect of U0126 on PMA- and dbcAMP/hCG-mediated StAR expression and steroid synthesis was tightly correlated with the expression of dosage-sensitive sex reversal, adrenal hypoplasia congenita, critical region on the X chromosome, gene 1 (DAX-1) and scavenger receptor class B type 1 (SR-B1). In fact, both DAX-1 and SR-B1 appear to play important roles in hormone-regulated steroidogenesis. These findings clearly demonstrate that the ERK1/2 signaling cascade involved in regulating StAR expression and steroid synthesis is mediated by multiple factors and pathways and is stimulus specific in mouse Leydig cells. PMID- 17400804 TI - Protection of pancreatic beta-cells by exendin-4 may involve the reduction of endoplasmic reticulum stress; in vivo and in vitro studies. AB - The aim of this study was to investigate the in vivo and in vitro effects of exendin-4, a potent glucagon-like peptide 1 agonist, on the protection of the pancreatic beta-cells against their cell death. In in vivo experiments, we used beta-cell-specific calmodulin-overexpressing mice where massive apoptosis takes place in their beta-cells, and we examined the effects of chronic treatment with exendin-4. Chronic and s.c. administration of exendin-4 reduced hyperglycemia. The treatment caused significant increases of the insulin contents of the pancreas and islets, and retained the insulin-positive area. Dispersed transgenic islet cells lived only shortly, and several endoplasmic reticulum (ER) stress related molecules such as immunoglobulin-binding protein (Bip), inositol requiring enzyme-1alpha, X-box-binding protein-1 (XBP-1), RNA-activated protein kinase-like endoplasmic reticulum kinase, activating transcription factor-4, and C/EBP-homologous protein (CHOP) were more expressed in the transgenic islets. We also found that the spliced form of XBP-1, a marker of ER stress, was also increased in beta-cell-specific calmodulin-overexpressing transgenic islets. In the quantitative real-time PCR analyses, the expression levels of Bip and CHOP were reduced in the islets from the transgenic mice treated with exendin-4. These findings suggest that excess of ER stress occurs in the transgenic beta-cells, and the suppression of ER stress and resultant protection against cell death may be involved in the anti-diabetic effects of exendin-4. PMID- 17400805 TI - Gradients in bone morphogenetic protein-related gene expression across the growth plate. AB - In the growth plate, stem-like cells in the resting zone differentiate into rapidly dividing chondrocytes of the proliferative zone and then terminally differentiate into the non-dividing chondrocytes of the hypertrophic zone. To explore the molecular switches responsible for this two-step differentiation program, we developed a microdissection method to isolate RNA from the resting (RZ), proliferative (PZ), and hypertrophic zones (HZ) of 7-day-old male rats. Expression of approximately 29,000 genes was analyzed by microarray and selected genes verified by real-time PCR. The analysis identified genes whose expression changed dramatically during the differentiation program, including multiple genes functionally related to bone morphogenetic proteins (BMPs). BMP-2 and BMP-6 were upregulated in HZ compared with RZ and PZ (30-fold each, P < 0.01 and 0.001 respectively). In contrast, BMP signaling inhibitors were expressed early in the differentiation pathway; BMP-3 and gremlin were differentially expressed in RZ (100- and 80-fold, compared with PZ, P < 0.001 and 0.005 respectively) and growth differentiation factor (GDF)-10 in PZ (160-fold compared with HZ, P < 0.001). Our findings suggest a BMP signaling gradient across the growth plate, which is established by differential expression of multiple BMPs and BMP inhibitors in specific zones. Since BMPs can stimulate both proliferation and hypertrophic differentiation of growth plate chondrocytes, these findings suggest that low levels of BMP signaling in the resting zone may help maintain these cells in a quiescent state. In the lower RZ, greater BMP signaling may help induce differentiation to proliferative chondrocytes. Farther down the growth plate, even greater BMP signaling may help induce hypertrophic differentiation. Thus, BMP signaling gradients may be a key mechanism responsible for spatial regulation of chondrocyte proliferation and differentiation in growth plate cartilage. PMID- 17400806 TI - Functional aspects of genetic variability in the GH genomic region. AB - Because of the small differences among genotypes, it would be difficult in basal conditions to detect the effect of genetic polymorphism in endocrine function, but this could emerge during provocative tests. We have studied four polymorphic sites of the GH gene region (17q24.2), MSPIA, MSPIB, BGLIIA, and BGLIIB. Gene and haplotype distributions in classes of growth retardation have been studied. The outcome of GH diagnostic test in relation to GH region genotypes has been evaluated by the analysis of area under the GH secretory curve. Ninety-eight growth retarded children have been studied. On the basis of provocative GH test these children were classified as total GH deficit (TD), partial GH deficit (PD), and familial short stature (FSS) with no deficit of GH. Sixty-three healthy controls were also considered. An increased frequency of MSPIA*2 allele in PD and TD as compared with FSS children and controls has been observed suggesting that this allele is associated with a decreased GH release. BGLIIA*2 allele appears decreased in PD and TD as compared with FSS and controls, suggesting that this allele is associated with an increased release of GH. Carriers of MSPIA*2 allele show a lower GH release as compared with MSPIA *1/*1 subjects on the provocative test by insulin, while carriers of BGLIIA*2 allele show a higher GH release as compared with BGLIIA *1/*1 subjects on the provocative test by clonidine. The functional aspects of genetic variability within the GH genomic area parallel the genetic differences observed between TD and PD versus FSS and control children. PMID- 17400807 TI - Overexpression of Wnt-1 in thyrocytes enhances cellular growth but suppresses transcription of the thyroperoxidase gene via different signaling mechanisms. AB - Wnt binding to cell surface receptors can activate a 'canonical' pathway that increases cellular beta-catenin or a 'noncanonical' Ca(++) pathway which can increase protein kinase C (PKC) activity. Although components of both Wnt/beta catenin-signaling pathways exist in thyrocytes, their biological role is largely unknown. In evaluating the biological role of Wnt signaling in differentiated FRTL-5 thyroid cells, we showed that TSH increased canonical Wnt-1 but, surprisingly, decreased the active form of beta-catenin. Transient overexpression of Wnt-1 or beta-catenin in FRTL-5 cells increased active beta-catenin (ABC), decreased thyroperoxidase (TPO) mRNA, and suppressed TPO-promoter activity. The target of beta-catenin suppressive action was a consensus T cell factor/lymphoid enhancing factor (TCF/LEF)-binding site 5'-A/T A/T CAAAG-3', -137 to -129 bp on the rat TPO promoter. beta-Catenin overexpression significantly increased complex formation between beta-catenin/TCF-1 and an oligonucleotide containing the TCF/LEF sequence, suggesting that the beta-catenin/TCF-1 complex acts as a transcriptional repressor of the TPO gene. Stable over-expression of Wnt-1 in FRTL-5 cells significantly increased the growth rate without increasing beta catenin levels. Increased growth was blunted by a PKC inhibitor, staurosporin. Wnt-1 overexpression increased serine phosphorylation, without affecting tyrosine phosphorylation, of signal transducers and activators of transcription 3 (STAT3) protein. In addition, these final results suggest that TSH-induced increase in Wnt-1 levels in thyrocytes contributes to enhanced cellular growth via a PKC pathway that increases STAT3 serine phosphorylation and activation, whereas TSH induced decrease in activation of beta-catenin simultaneously relieves transcriptional suppression of TPO. We hypothesize that Wnt signaling contributes to the ability of TSH to simultaneously increase cell growth and functional, thyroid-specific, gene expression. PMID- 17400808 TI - The integrated action of oestrogen receptor isoforms and sites with progesterone receptor in the gonadotrope modulates LH secretion: evidence from tamoxifen treated ovariectomized rats. AB - The specific role of each oestrogen receptor (ER) isoform (alpha and beta ) and site (nucleus and plasma membrane) in LH release was determined in ovariectomized (OVX) rats injected over 6 days (days 15-20 after OVX) with a saturating dose (3 mg/day) of tamoxifen (TX), a selective ER modulator with nuclear ERalpha agonist actions in the absence of oestrogen. This pharmacological effect of TX was demonstrated by the fact that it was blocked by the selective ERalpha antagonist methyl-piperidinopyrazole. Over the past 3 days of the 6-day TX treatment, rats received either 25 microg/day oestradiol benzoate (EB), 1.5 mg/day selective ERalpha agonist propylpyrazole triol (PPT) and the selective ERbeta agonist diarylpropionitrile (DPN), or a single 3 mg injection of the antiprogestin onapristone (ZK299) administered on day 20. Blood samples were taken to determine basal and progesterone receptor (PR)-dependent LH-releasing hormone (LHRH) stimulated LH secretion and to evaluate LHRH self-priming, the property of LHRH that increases gonadotrope responsiveness to itself. Blood LH concentration was determined by RIA and gonadotrope PR expression by immunohistochemistry. Results showed that i) EB and DPN potentiated the negative feedback of TX on basal LH release; ii) DPN reduced TX-induced PR expression; iii) EB and PPT blocked TX elicited LHRH self-priming and iv) ZK299 reduced LHRH-stimulated LH secretion and blocked LHRH self-priming. These observations suggest that oestrogen action on LH secretion in the rat is exerted at the classic ERalpha pool and that this action might be modulated by both ERbeta and membrane ERalpha through their effects on PR expression and action respectively. PMID- 17400809 TI - Expression of the oxytocin gene, but not the vasopressin gene, in the rat uterus during pregnancy: influence of oestradiol and progesterone. AB - Oxytocin (OT) and vasopressin (VP) are neurohypophyseal hormones with potent stimulatory actions on the uterus. In order to determine whether these hormones may have a paracrine action on the uterus, OT and VP gene expression was studied in myometrium from pregnant rats at gestational ages of 14 and 20 days, and from ovariectomized animals treated with oestradiol and progesterone. OT and VP mRNA concentrations were measured using real-time quantitative reverse transcription PCR, and OT- and VP-like immunoreactivities were determined using RIA. OT mRNA was detected in the uterus from pregnant rats, but did not differ between the groups of different gestational ages. Oestradiol significantly (P<0.05) stimulated OT gene expression in ovariectomized rats. Progesterone alone was without effect on OT mRNA concentrations, but significantly (P<0.05) reduced the oestradiol-induced OT mRNA accumulation. The OT-like immunoreactivity in an extract of myometrium from pregnant rats was eluted from a reverse-phase HPLC column with a retention time identical to that of synthetic OT. Neither VP mRNA nor VP-like immunoreactivity was detected in the myometrium from pregnant or ovariectomized rats. The study demonstrates steroid-dependent expression of the OT gene in the rat uterus and processing of uterine preprooxytocin to the mature nonapeptide. The data support the theory that this peptide may act in a paracrine pathway. No evidence was found for the presence of VP in the uterus so that, if the hormone is involved in a stimulatory action on this tissue, it probably acts via an endocrine mechanism. PMID- 17400810 TI - The role of glucocorticoid in the regulation of prostaglandin biosynthesis in non pregnant bovine endometrium. AB - To determine whether glucocorticoids (GCs) play a role in regulating uterine function in cow, the present study examined the expression of mRNA encoding GC receptor (GC-R) alpha, 11beta-hydroxysteroid dehydrogenase (11-HSD) type 1 and type 2, and the activity of 11-HSD1 in bovine endometrial tissue throughout the estrous cycle. We also studied the effects of cortisol on basal, oxytocin (OT)- and tumor necrosis factor-alpha (TNFalpha)-stimulated prostaglandin (PG) production. A quantitative real-time PCR analysis revealed that GC-Ralpha mRNA was expressed more strongly in the mid-luteal stage (days 8-12) than in the other stages. In contrast to GC-Ralpha mRNA expression, 11-HSD1 mRNA expression was greater in the follicular stage than in the other stages, whereas 11-HSD2 mRNA expression was lowest in the follicular stage. The activity of 11-HSD1 was greater in the follicular stage and estrus than in the other stages and was lowest in the mid-luteal stage. Cortisone was dose-dependently converted to cortisol in the cultured endometrial tissue. Although cortisol did not affect either the basal or OT-stimulated production of PGs in the cultured epithelial cells, the production of PGs stimulated by TNFalpha in the stromal cells was suppressed by cortisol (P < 0.05). Cortisol suppressed basal prostaglandin (PG)F2alpha without affecting basal PGE2 production in the stromal cells. The overall results suggest that the level of cortisol is locally regulated in bovine endometrium throughout the estrous cycle by 11-HSD1, and that cortisol could act as a luteoprotective factor by selectively suppressing luteolytic PGF2alpha production in bovine endometrium. PMID- 17400811 TI - Local adiponectin treatment reduces atherosclerotic plaque size in rabbits. AB - In this study, we investigated the in vivo role of adiponectin, an adipocytokine, on the development of atherosclerosis in rabbits mainly using adenovirus expressing adiponectin gene (Ad-APN) and intravascular ultrasonography. Serum adiponectin concentrations in rabbits after Ad-APN local transfer to abdominal aortas increased about nine times as much as those before transfer (P < 0.01), about ten times as much as the levels of endogenous adiponectin in adenovirus expressing beta-galactosidase gene (Ad-beta gal) treated rabbits (P < 0.01), and about four times as much as those in the aorta of non-injured rabbits on a normal cholesterol diet (P < 0.01). Ultrasonography revealed a significantly reduced atherosclerotic plaque area in abdominal aortas of rabbits infected through intima with Ad-APN, by 35.2% compared with the area before treatment (P < 0.01), and by 35.8% compared with that in Ad-beta gal-treated rabbits (P < 0.01). In rabbits infected through adventitia, Ad-APN treatment reduced plaque area by 28.9% as compared with the area before treatment (P < 0.01) and 25.6% compared with that in Ad-beta gal-treated rabbits (P < 0.01). Adiponectin significantly suppressed the mRNA expression of vascular cell adhesion molecule-1 (VCAM-1) by 18.5% through intima transfer (P < 0.05) and 26.9% through adventitia transfer (P < 0.01), and intercellular adhesion molecule-1 (ICAM-1) by 40.7% through intima transfer (P < 0.01), and 30.7% through adventitia transfer (P < 0.01). However, adiponectin had no effect on the expression of types I and III collagen. These results suggest that local adiponectin treatment suppresses the development of atherosclerosis in vivo in part by attenuating the expression of VCAM-1 and ICAM 1 in vascular walls. PMID- 17400812 TI - Coactivator-mediated estrogen response in human squamous cell carcinoma lines. AB - Steroid hormones such as 17beta-estradiol (E2) are critical to diverse cellular processes including tumorigenesis. A number of cofactors such as nuclear receptor corepressor (NCoR), CREB-binding protein (CBP), and steroid receptor coactivator 1 (SRC-1) interact with estrogen receptors (ERs) to regulate transcriptional repression or activation of target genes. Estrogen signaling in non-reproductive tract tissues such as skin is less well characterized and the effectiveness of anti-estrogen therapy for cancer arising from these tissues is unknown. We show that tamoxifen (TAM) treatment inhibited cell cycle progression and proliferation of human cancer lines derived from stratified squamous epithelium squamous cell carcinoma (SCC). E2 had no effect on proliferation of these lines despite low levels of ERalpha expression. The E2 treatment promoted displacement of the NCoR from ERalpha and recruitment of CBP to the receptor. SRC-1 expression was not detected in these SCC lines; however, transient transfection of SRC-1, CBP, or both coactivators enhanced transactivation of an estrogen responsive promoter in cancer cells treated with E2 or TAM. In stable clones expressing SRC-1, the coactivator was recruited to ERalpha along with CBP in E2 but not in TAM-treated cells. SRC-1 expression restored the E2-mediated proliferative response to human SCC lines. This increased proliferation correlated with increased extracellular signal regulated kinase 1 (ERK1) expression. SRC-1 and CBP were recruited to the proximal ERK1 promoter region in E2 but not in TAM-treated cells. We concluded that SRC-1 was a key molecular determinant of estrogen-mediated proliferation in human SCC lines. PMID- 17400813 TI - Expression of molecular equivalent of hypothalamic-pituitary-adrenal axis in adult retinal pigment epithelium. AB - We have investigated expression of molecular elements of the hypothalamic pituitary-adrenal (HPA) axis in the human retinal pigment epithelium (RPE) cells. The presence of corticotropin-releasing factor (CRF); urocortins I, II and III; CRF receptor type 1 (CRFR1); POMC and prohormone convertases 1 and 2 (PC1 and PC2) mRNAs were shown by RT-PCR; the protein products were detected by ELISA, western blot or immunocytochemical methods in an ARPE-19 cell line derived from an adult human donor. CRFR2 was below the level of detectability. The CRFR1 was functional as evidenced by CRF stimulation of cAMP and inositol triphosphate production as well as by ligand induction of transcriptional activity of inducible cis-elements cAMP responsive element (CRE), activator protein 1 responsive element (AP-1) and POMC promoter) in ARPE-19 using luciferase reporter assay. Immunoreactivities representative of CRF, pre-urocortin, CRFR1 receptor and ACTH were also detected in mouse retina by in situ immunocytochemistry. Finally, using RT-PCR, we detected expression of genes encoding four key enzymes participating in steroids synthesis (CYP11A1, CYP11B1, CYP17 and CYP21A2) and showed transformation of progesterone into cortisol-immunoreactivity in cultured ARPE-19 cells. Therefore, we suggest that ocular tissue expresses CRF-driven signalling system that follows organisational structure of the HPA axis. PMID- 17400814 TI - Osteopontin deficiency enhances anabolic action of EP4 agonist at a sub-optimal dose in bone. AB - Osteoporosis is one of the most widespread and destructive bone diseases in our modern world. There is a great need for anabolic agents for bone which could reverse this disease, but few are available for clinical use. Prostaglandin E receptor (EP4) agonist (EP4A) is one of the very few anabolic agents for bone in rat, but its systemic efficacy against bone loss at sub-optimal dose is limited in mice. As osteoblasts are regulated by extracellular matrix proteins, we tested whether deficiency of osteopontin (OPN), a secreted phosphorylated protein, could modulate the effects of EP4A (ONO-AE1-329) treatment at 30 microg/kg body weight, a sub-optimal dose, for 5 days/week for 4 weeks. OPN deficiency enhanced the anabolic effects of EP4A on bone volume. Histomorphometric analysis indicated that EP4A increased mineral apposition rate as well as bone formation rate in OPN deficient but not in wild-type mice. Neither OPN deficiency nor EP4A altered osteoclast parameters. Importantly, OPN deficiency enhanced the direct anabolic action of EP4A locally injected onto the parietal bone in inducing new bone formation. Combination of OPN deficiency and EP4A treatment caused an increase in mineralized nodule formation in the cultures of bone marrow cells. Finally, OPN deficiency enhanced anabolic action of EP4A in the mice subjected to ovariectomy. These data indicate that OPN deficiency enhances the actions of EP4A at sub optimal dose. PMID- 17400815 TI - Cloning and characterization of the ecdysone receptor and ultraspiracle protein from the water flea Daphnia magna. AB - cDNAs encoding the ecdysone receptor (EcR) and ultra spiracle (USP) protein were cloned from the water flea Daphnia magna (Crustacea: Cladocera). The deduced EcR and USP amino acid sequences showed a high degree of homology to those of other crustaceans as well as insects. We isolated three isoforms of EcR that differ in the A/B domain. Quantitative PCR analysis indicated differing temporal expression patterns of the EcR isoforms during the molting period and demonstrated that the expression of one subtype correlated well with the timing of molt. Using cDNAs encoding EcR and USP, we constructed a Daphnia EcR/USP reporter based on a two hybrid system. The gene fusions encoded the EcR ligand-binding domain (LBD) fused to the Gal4 DNA-binding domain, and the USP-LBD fused to the Vp16 activation domain. These chimeric genes were transfected with a luciferase reporter gene. Dose-dependent activation of the reporter gene could be observed when transfectants were exposed to Ec and other chemicals known to have Ec-like activities. This two-hybrid system may represent a useful reporter system for further examination of hormonal and chemical effects on Daphnia at the molecular level. PMID- 17400816 TI - Structure probing of tmRNA in distinct stages of trans-translation. AB - Ribosomes stalled on problematic mRNAs in bacterial cells can be rescued by transfer-messenger RNA (tmRNA), its helper protein (small protein B, SmpB), and elongation factor Tu (EF-Tu) through a mechanism called trans-translation. In this work we used lead(II) footprinting to probe the interactions of tmRNA with SmpB and other components of the translation machinery at different steps of the trans-translation cycle. Ribosomes with a short nascent peptide stalled on a truncated mRNA were reacted with Ala-tmRNA*EF-Tu*GTP, SmpB, and other translation components to initiate and execute trans-translation. Free tmRNA was probed with lead(II) acetate with and without SmpB, and ribosome bound tmRNA was probed in one of four different trans-translation states stabilized by antibiotic addition or selective exclusion of translation components. For comparison, we also analyzed lead(II) cleavage patterns of tmRNA in vivo in a wild-type as well as in an SmpB-deficient Escherichia coli strain. We observed some specific cleavages/protections in tmRNA for the individual steps of trans-translation, but the overall tmRNA conformation appeared to be similar in the stages analyzed. Our findings suggest that, in vivo, a dominant fraction of tmRNA is in complex with SmpB and that, in vitro, SmpB remains tmRNA bound at the initial steps of trans translation. PMID- 17400817 TI - Double-stranded regions are essential design components of potent inhibitors of RISC function. AB - While microRNAs (miRNAs) are recognized as playing a critical role in regulating eukaryotic gene expression, both the mechanism by which these small, noncoding RNAs function and the genes they target remain elusive. Previous studies have shown that short, single-stranded 2'-O-methyl-modified oligonucleotides that are complementary to mature microRNA sequences can interact with the miRNA-RISC nucleoprotein complex and weakly inhibit miRNA function. Here we report the identification of secondary structural elements that enhance the potency of these molecules. Incorporation of highly structured, double-stranded flanking regions around the reverse complement core significantly increases inhibitor function and allows for multi-miRNA inhibition at subnanomolar concentrations. The improved functionality of these double-stranded miRNA inhibitors may provide insights into the miRNA mechanism by suggesting the possible importance of such structures in or near endogenous miRNA target sites. PMID- 17400818 TI - Bunyavirus mRNA synthesis is coupled to translation to prevent premature transcription termination. AB - Messenger RNA transcription by Bunyaviridae family members is unique within the group of negative-strand RNA viruses as it requires on-going protein synthesis. The long-standing model explaining this phenomenon proposes that the translational requirement is not for a protein product, but instead is for ribosomal translocation along nascent mRNAs. This movement is proposed to disrupt spurious transcription termination signals that otherwise cause premature mRNA truncation leading to a fatal loss of gene expression. This model was tested by introducing translational stop codons into model RNA genomes of Bunyamwera virus, the prototypic member of the Bunyaviridae family. This directly showed that translation of nascent mRNAs prevents transcription termination. While such coupling of transcription and translation is common in prokaryotic systems, these results represent the first report of such obligatory coupling in a eukaryotic cell environment. The results also provide insight into the bunyavirus termination mechanism and suggest it is mechanistically similar to prokaryotic intrinsic termination. PMID- 17400819 TI - Immediate early response of the circadian polyA ribonuclease nocturnin to two extracellular stimuli. AB - Nocturnin (Noc, also called Ccrn4l [carbon catabolite repression 4-like]) is a circadian deadenylase that is rhythmically expressed in multiple tissues in mice with peak mRNA levels in early night. Since several other circadian genes are induced by extracellular stimuli, we tested the hypothesis that Noc is acutely regulated in NIH3T3 cells. A serum shock and the phorbol ester TPA induced Noc transcript levels in quiescent NIH3T3 cultures while dexamethasone and forskolin, which are known to induce other clock genes in culture, were without effect. NOC protein levels also were induced by serum. The half-life of the TPA-induced Noc mRNA is short, and the inhibition of protein synthesis by cycloheximide prevents Noc mRNA degradation and revealed a 30-fold increase in the transcript levels after 4 h of TPA treatment. Since this acute induction is not dependent on protein synthesis, Noc behaves like other immediate early genes. Remarkably, these acute effects are specific to Noc as the mRNAs encoding other known mouse deadenylases, CCR4, CAF1, PAN2, and PARN, were not induced in the same paradigm. Our data show that in addition to its robust circadian regulation, Noc expression can be regulated acutely, and imply that it can respond directly and specifically to physiological cues. NOC may act in turning off the expression of genes that are required to be silenced as a response to these extracellular signals. PMID- 17400820 TI - The effect of multiple doses of peginterferon alfa-2b on the steady-state pharmacokinetics of methadone in patients with chronic hepatitis C undergoing methadone maintenance therapy. AB - This multicenter, open-label study evaluated the effects of multiple doses of peginterferon alfa-2b on the steadystate pharmacokinetics of methadone in 20 adults with hepatitis C virus infection who were enrolled in a methadone maintenance program. All subjects received peginterferon alfa-2b 1.5 mug/kg/wk for 4 weeks and maintained their normal methadone regimen. Serial blood samples were collected immediately before the first and after the fourth peginterferon alfa-2b dose (day 23). At day 23, exposure to the active methadone R-enantiomer increased by approximately 15% following administration of peginterferon alfa-2b, with 90% confidence intervals just outside the bioequivalence criteria (range, 80%-125%). Similar increases in exposure (C(max), AUC(0-24), and AUC(last)) were observed with S-methadone and total methadone. Peginterferon alfa-2b was well tolerated. Peginterferon alfa-2b is associated with minor increases in exposure to methadone in individuals with hepatitis C virus infection; however, these increases are unlikely to be clinically meaningful and are not associated with any safety concerns. PMID- 17400821 TI - Complementary, holistic, and integrative medicine: chamomile. PMID- 17400822 TI - Heart murmurs. PMID- 17400823 TI - Presentation of congenital heart disease in the neonate and young infant. PMID- 17400824 TI - Evidence-based medicine: critical appraisal of the literature (critical appraisal tools). PMID- 17400825 TI - Index of suspicion. PMID- 17400826 TI - Visual diagnosis: an adolescent who has fever, joint pain, a rash, and "blacking out" episodes. PMID- 17400829 TI - Pediatrics in the community: taking a collective breath to help children who have asthma. PMID- 17400830 TI - Alcohol. PMID- 17400831 TI - Marijuana. PMID- 17400833 TI - Power, risk, and the status quo: does power promote riskier or more conservative decision making? AB - Two experiments suggest that the experience of power can interact with a person's level of power motivation to produce effects on risky decision making. In Study 1, assignment to a position of power increased risk taking among participants with low levels of power motivation but reduced risk taking among participants with high levels of power motivation. In Study 2, participants high in power motivation again made more conservative decisions, but only under circumstances in which the dominance hierarchy was unstable and there was potential for losing their power. When power was irrevocable and participants' choices had no bearing on their ability to retain power, both high and low power-motivated participants responded by making riskier decisions. Findings suggest that although power may generally lead to riskier decisions, power may lead to more conservative decisions among power-motivated individuals, especially when the status quo is perceived to be in jeopardy. PMID- 17400834 TI - Labor, delivery, and early parenthood: an attachment theory perspective. AB - Having a baby is a powerful experience that, according to attachment theory, should activate the attachment system and elicit attachment needs and motives. The current study investigated first-time parents' perceptions of and responses to events surrounding labor and delivery and early parenthood. Couples expecting their first child completed measures of attachment orientations and questionnaires assessing key aspects of the experience of labor, delivery, and early parenthood. Attachment anxiety and avoidance significantly predicted individuals' perceptions of themselves and their partners, reactions to their partners' behavior, and emotional responses to their infants. Jealousy of the infant, perceptions of support given to or received from partners, and feelings of closeness to the infant were all associated with attachment orientations in theoretically meaningful ways. PMID- 17400835 TI - Unmitigated communion and health among adolescents with and without diabetes: the mediating role of eating disturbances. AB - The authors examined the implications of unmitigated communion--a focus on others to the exclusion of the self--for psychological and physical health among adolescents with (n = 132) and without (n = 131) diabetes times/during a 1-year period. Unmitigated communion predicted greater psychological distress and lower levels of competence cross-sectionally and longitudinally, controlling for sex, pubertal status, and communion. Health status moderated some longitudinal relations, such that relations held only for adolescents with diabetes. In addition, unmitigated communion was associated with poor metabolic control and predicted a deterioration in metabolic control over the year for adolescents with diabetes. Unmitigated communion also was associated with disturbed eating behavior cross-sectionally and longitudinally, and disturbed eating behavior explained some of the relations of unmitigated communion to psychological but not physical health outcomes. PMID- 17400836 TI - Why individuals protest the perceived transgressions of their country: the role of anger, shame, and guilt. AB - The present research examined emotions as predictors of opposition to policies and actions of one's country that are perceived to be illegitimate. Two studies investigated the political implications of American (Study 1) and British (Study 2) citizens' anger, guilt, and shame responses to perceived harm caused by their countries' occupation of Iraq. In both studies, a manipulation of pervasive threat to the country's image increased participants' shame but not guilt. The emotions predicted political action intentions to advocate distinct opposition strategies. Shame predicted action intentions to advocate withdrawal from Iraq. Anger predicted action intentions to advocate compensation to Iraq, confrontation of agents responsible, and withdrawal from Iraq. Anger directed at different targets (ingroup, ingroup representative, and outgroup representative) predicted action intentions to support distinct strategies (Study 2). Guilt did not independently predict any political action intentions. Implications for the study of political action and emotions in intergroup contexts are discussed. PMID- 17400837 TI - Vaginal birth after cesarean prediction: a self-fulfilling prophecy? PMID- 17400838 TI - Gestational age: not always what it seems. PMID- 17400839 TI - Predicting failure of a vaginal birth attempt after cesarean delivery. AB - OBJECTIVE: To identify a group of clinical factors that could be used to accurately predict failure in women attempting vaginal birth after cesarean (VBAC). METHODS: We conducted a planned secondary analysis of a retrospective cohort study of women who were offered VBAC from 1996 to 2000 in 17 community and university hospitals. We collected information about maternal history and outcomes of the index pregnancy. We used univariable and multivariable statistical methods to develop a multivariable prediction model for the outcome of VBAC failure. RESULTS: A total of 13,706 patients attempted VBAC, with a failure rate of 24.5%. Six variables were significantly associated with VBAC failure in our final logistic regression model: gestational age at delivery, maternal age, maternal race, labor type (spontaneous, augmented, or induced), history of vaginal delivery, and cephalopelvic disproportion or failed induction (combined variable) as prior cesarean indication. The area under the receiver operating characteristics curve is 0.717. To achieve a sensitivity of approximately 75%, a false-positive rate of approximately 40% would result. CONCLUSION: Our results indicate that significant clinical variables (prelabor and labor) cannot reliably predict VBAC failure. LEVEL OF EVIDENCE: II. PMID- 17400840 TI - Development of a nomogram for prediction of vaginal birth after cesarean delivery. AB - OBJECTIVE: To develop a model based on factors available at the first prenatal visit that predicts chance of successful vaginal birth after cesarean delivery (VBAC) for individual patients who undergo a trial of labor. METHODS: All women with one prior low transverse cesarean who underwent a trial of labor at term with a vertex singleton gestation were identified from a concurrently collected database of deliveries at 19 academic centers during a 4-year period. Using factors identifiable at the first prenatal visit, we analyzed different classification techniques in an effort to develop a meaningful prediction model for VBAC success. After development and cross-validation, this model was represented by a graphic nomogram. RESULTS: Seven-thousand six hundred sixty women were available for analysis. The prediction model is based on a multivariable logistic regression, including the variables of maternal age, body mass index, ethnicity, prior vaginal delivery, the occurrence of a VBAC, and a potentially recurrent indication for the cesarean delivery. After analyzing the model with cross-validation techniques, it was found to be both accurate and discriminating. CONCLUSION: A predictive nomogram, which incorporates six variables easily ascertainable at the first prenatal visit, has been developed that allows the determination of a patient-specific chance for successful VBAC for those women who undertake trial of labor. LEVEL OF EVIDENCE: II. PMID- 17400841 TI - Reconciling the high rates of preterm and postterm birth in the United States. AB - OBJECTIVE: Preterm and postterm birth rates are substantially higher in the United States than in Canada and other industrialized countries, although relative mortality at preterm compared with term gestation is considerably lower. We attempted to explain these differences based on differences in the method of gestational age estimation. METHODS: We used information on all live births in the United States and Canada for 1995-2002 and on singleton births and perinatal deaths for 1996-1999. Gestational age in Canada was based on the clinical estimate, whereas in the United States both menstrual-based and clinical estimates were used. RESULTS: In 2002, preterm (12.3%) and postterm birth (6.6%) rates in the United States were far higher than in Canada (7.6% and 1.0%, respectively) when U.S. rates were based on menstrual dates. Differences were reduced or abolished when U.S. rates were based on the clinical estimate of gestation (10.1% and 1.0%, respectively). In Canada, the rate ratio for perinatal death at preterm compared with term gestation was 27.8 (95% confidence interval [CI] 26.3-29.3), similar to that in the United States when gestation was based on the clinical estimate (rate ratio 26.5, 95% CI 26.1-26.9, P value for difference in rate ratios=.06) but not when based on menstrual dates (rate ratio 18.9, 95% CI 18.7-19.2, P<.001). CONCLUSION: Menstrual dates in U.S. data misclassify gestational duration and overestimate both preterm and postterm birth rates. For international comparisons, gestational age in the United States should be based on the clinical estimate. LEVEL OF EVIDENCE: II. PMID- 17400842 TI - Ineffectiveness of sertraline for treatment of menopausal hot flushes: a randomized controlled trial. AB - OBJECTIVE: To estimate the effect of the selective serotonin reuptake inhibitor sertraline on hot flush frequency and severity in perimenopausal and postmenopausal women. METHODS: We performed a randomized, blinded, placebo controlled trial in women aged 40 to 60 years with 14 or more hot flushes per week (N=99). Women were randomly assigned initially to daily oral sertraline (50 mg) or identical placebo for 2 weeks. If no substantial side effects were noted, the dose was increased to two tablets daily (100 mg sertraline or placebo) and continued for an additional 4 weeks. Hot flush frequency and severity were recorded on a daily diary. Hot flush score was calculated as frequency multiplied by severity. Participants also completed questionnaires addressing quality of life, menopausal symptoms, sleep quality, sexual function, mood, and side effects. RESULTS: After 6 weeks of treatment, hot flush frequency decreased similarly in both the placebo (38%) and sertraline (39%) groups (P=.94). Mean hot flush scores also decreased similarly in both groups (41% and 42%, respectively, P=.86). Compared with placebo, women in the sertraline group were more likely to report gastrointestinal complaints, dry mouth, and dizziness. Treatment with sertraline also resulted in greater worsening of scores on the Medical Outcomes Study (MOS) Short Form 36 standardized physical component and the global Female Sexual Function Index. Results were similar in women at least 80% adherent to study medication. CONCLUSION: Treatment with sertraline did not improve hot flush frequency or severity in generally healthy perimenopausal and postmenopausal women, but was associated with bothersome side effects. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00283192 LEVEL OF EVIDENCE: I. PMID- 17400843 TI - Hormones and sexuality during transition to menopause. AB - OBJECTIVE: To examine the relationship between reproductive hormonal dynamics and sexual dysfunction assessed in a cohort of women approaching menopause. METHODS: Women in the Penn Ovarian Aging Study were assessed at yearly intervals for 3 years with early follicular hormone measurements (estradiol, follicle-stimulating hormone, luteinizing hormone [LH], sex hormone binding globulin, dehyroepiandrosterone sulfate [DHEAS], total testosterone), anthropometric measures, and extensive questionnaires including the Female Sexual Function Index. Univariable analyses were performed to determine the association between hormones, menopausal status, and sexual dysfunction. Multivariable linear and logistic regression models were created to examine the influence of hormones on sexual function adjusting for the effect of potential confounders. RESULTS: The final multivariable model indicated that sexual dysfunction increased with advanced menopausal status, with postmenopausal women being 2.3 times as likely to experience sexual dysfunction compared with premenopausal women (odds ratio 2.3, 95% confidence interval [CI] 1.3-4.1). Low DHEAS serum concentrations were associated with decreased sexual function (odds ratio 1.59, 95% CI 1.19-2.14). Additional risk factors associated with sexual dysfunction included absence of a sexual partner (11.2, 95% CI 6.9-18.1), high anxiety (3.8, 95% CI 1.6-9.2), and children under the age of 18 living at home (1.6, 95% CI 1.1-5.5). Lubrication, orgasm, and pain were specific aspects of sexuality negatively affected by menopause. CONCLUSION: This study confirms the observation that sexual dysfunction increases over the menopausal transition. Several factors associated with sexual dysfunction include low DHEAS, absence of a sexual partner, anxiety, and children under the age of 18 living at home. LEVEL OF EVIDENCE: II. PMID- 17400844 TI - Hot flushes, bone mineral density, and fractures in older postmenopausal women. AB - OBJECTIVE: To assess whether greater severity of hot flushes is associated with bone loss or fracture risk in older postmenopausal women. METHODS: This study is a secondary analysis of 3,167 postmenopausal women in the Multiple Outcomes of Raloxifene Evaluation trial. Baseline hot flush severity was assessed by self report. Femoral neck and lumbar spine bone density was assessed by dual-energy X ray absorptiometry. Vertebral and nonvertebral fractures were determined radiographically and by interview. Baseline bone density, 3-year bone loss, baseline prevalent fractures, and 3-year fracture incidence were examined in women with varying hot flush severity. RESULTS: After adjusting for other characteristics, greater severity of hot flushes was associated with higher, rather than lower, baseline bone density (adjusted mean femoral neck bone density=0.633, 95% confidence interval [CI] 0.614-0.652 g/cm2, versus 0.611, 95% CI 0.608-0.613 g/cm2 for women with "severe" versus "minimal" hot flushes). Women with more severe hot flushes were less likely to have a baseline fracture (odds ratio 0.64, 95% CI 0.48-0.84, for vertebral fracture in women with moderate or severe versus minimal hot flushes). The 3-year annualized change in bone density did not differ among women with varying hot flush severity (P>.40 for all). Hot flush severity was not related to incident vertebral or nonvertebral fracture (P>.05 for all). CONCLUSION: Among osteoporotic women who are 5 or more years postmenopausal, greater severity of persistent hot flushes is not associated with progressive bone loss or risk of fracture, despite previous research linking hot flushes to bone loss during early menopause. LEVEL OF EVIDENCE: II. PMID- 17400845 TI - Natural history of pelvic organ prolapse in postmenopausal women. AB - OBJECTIVE: To describe the natural history of pelvic organ prolapse and risk factors for changes in vaginal descent in older women. METHODS: This 4-year prospective observational study included 259 postmenopausal women with a uterus enrolled at one Women's Health Initiative clinical site who completed at least two annual pelvic organ prolapse quantification (POP-Q) examinations. We calculated 1-year and 3-year incidence and resolution risks for prolapse (defined as maximal vaginal descent to or beyond the hymen) and estimated progression and regression rates (1 cm or greater and 2 cm or greater changes in maximal vaginal descent) and risk factors. RESULTS: Mean age was 68.1+/-5.5 years, and median vaginal parity was 4. Seventy-three (28%) women had four exams, 128 (49%) had three exams, and 58 (22%) had two exams. Prolapse waxed and waned yearly in individual women. Overall 1-year and 3-year prolapse incidences were 26% (95% confidence interval [CI] 20-33%) and 40% (95% CI 26-56%); 1-year and 3-year prolapse resolution risks were 21% (95% CI 11-33%) and 19% (95% CI 7-39%). Rates of any change in maximal vaginal descent over time varied depending on baseline measurements. Over 3 years, the maximal vaginal descent increased by at least 2 cm in 11.0% (95% CI 4.9-20.5%) of the women and decreased by at least 2 cm in 2.7% (95% CI 0.3-9.5%). Increasing body mass index and grand multiparity increased the risk for vaginal descent progression. CONCLUSION: Prolapse progresses and regresses in older women, although rates of vaginal descent progression are slightly greater than regression overall. Obesity is a risk factor for progression in vaginal descent. LEVEL OF EVIDENCE: III. PMID- 17400846 TI - Fetal adrenal gland volume: a novel method to identify women at risk for impending preterm birth. AB - OBJECTIVE: To investigate the risk of preterm birth (delivery at less than 37 weeks of gestation) by evaluating the fetal adrenal gland volume, hallmark of activation of the fetal hypothalamic-pituitary-adrenal axis, measured by 3 dimensional ultrasonography. METHODS: We performed 3-dimensional ultrasound examination of the fetal adrenal gland volume in 126 singleton fetuses, prospectively comparing those born to mothers with signs or symptoms of preterm labor (n=53) to control subjects (n=73). Multiplanar technique with rotational methods for measurement of fetal adrenal gland volume was performed by using Virtual Organ Computer-Aided Analysis (VOCAL) technology. RESULTS: The fetal adrenal gland volume was successfully examined in 86.5% of the cases. There was a direct relationship between the fetal adrenal gland volume and estimated fetal weight. A corrected adrenal gland volume of greater than 422 mm3/kg was best in predicting preterm birth within 5 days from the time of the measurement. The sensitivity, specificity, and positive and negative likelihood ratios were 92%, 99%, 93.5, and 0.08, respectively. Multiple logistic regression analysis showed that the corrected adrenal gland volume was the only significant independent predictor factor of preterm birth within 5 days of measurement. CONCLUSION: Corrected adrenal gland volume measurement may identify women at risk for impending preterm birth. This information can be generated noninvasively and in time for clinical decision making. LEVEL OF EVIDENCE: II. PMID- 17400847 TI - Natural history of cervical funneling in women at high risk for spontaneous preterm birth. AB - OBJECTIVE: To estimate the natural history of funneling in the second trimester by transvaginal ultrasonograms and whether funneling increases the risk of spontaneous birth. METHODS: Secondary analysis of a blinded, multi-center observational study of women with at least one prior spontaneous preterm birth at 16.0-31.9 weeks who subsequently carried singleton gestations. Cervical length, funneling (membrane prolapse greater than or equal to 5 mm), funnel shape, and dynamic changes were recorded at 16-18 weeks, and then every 2 weeks until 23.9 weeks. Managing obstetricians were blinded to the ultrasonography results. The primary outcome was gestational age at delivery. RESULTS: Five hundred ninety scans were performed in 183 women, of which 60 (33%) had funneling observed on at least one of the serial evaluations. These 60 women delivered at an earlier gestational age at delivery than the 123 women without funneling (31.7+/-7.9 weeks compared with 36.9+/-4.4 weeks; P<.001). In the 60 women with funneling on at least one evaluation, the progression over time of internal os cervical anatomy from a "T" to a "V" to a "U" shape was associated with earlier gestational age at delivery, whereas resolution of "V" shape funnels was associated with term delivery. Women with a shortened cervical length less than 25 mm (n=60) had a similar gestational age at birth with or without funneling (30.6+/-8.0 weeks compared with 31.9+/-6.6 weeks; P=.59). After controlling for the shortest observed cervical length, largest funnel percent was not a significant independent risk factor. CONCLUSION: The natural history of second trimester funneling has significant variability and a significant association with earlier gestational age at delivery. As an independent finding, funneling does not add appreciably to the risk of early gestational age at delivery associated with a shortened cervical length. LEVEL OF EVIDENCE: II. PMID- 17400848 TI - Presence of the sickle cell trait and preterm delivery in African-American women. AB - OBJECTIVE: To estimate the association between presence of the sickle cell trait and preterm delivery among African-American women. METHODS: A retrospective cohort study was conducted to study all deliveries by African-American women at one institution between 1976 and 2001. The primary predictor of interest was the presence of the sickle cell trait, and the primary outcome was preterm delivery. Post hoc analyses were conducted to explore the relationship between presence of sickle cell trait and multiple gestations, an apparent modifier of the relationship between sickle cell trait and preterm delivery. RESULTS: Of the 5,028 African-American women eligible for inclusion, 6.5% carried the sickle cell trait. In adjusted analyses, the presence of sickle cell trait was associated with a lower risk of preterm delivery at less than 32 weeks (adjusted odds ratio 0.15, 95% confidence interval 0.05-0.49), and was found to be associated with an increased odds of multiple gestations (adjusted odds ratio 1.94, 95% confidence interval 1.22-2.09). A significant interaction exists between the presence of multiple gestation and sickle cell trait and the odds of preterm delivery, such that the protective effect of sickle cell trait on preterm delivery risk is greater among those with multiple gestations than those with singletons. CONCLUSION: Among African-American women, the presence of the sickle cell trait was associated with lower odds of early preterm delivery despite adjustment for potential confounders. An increased odds of multiple gestations was also noted among these women, possibly suggesting a persistence of selection for the hemoglobin S trait in nonmalarial settings. LEVEL OF EVIDENCE: II. PMID- 17400849 TI - Mental disorders and nicotine dependence among pregnant women in the United States. AB - OBJECTIVE: To investigate the association between mental disorders and cigarette use and nicotine dependence among pregnant women in the United States. METHODS: A face-to-face general population survey was conducted on participants in the 2001 2002 National Epidemiologic Survey on Alcohol and Related Conditions. One thousand five hundred sixteen women reporting a pregnancy in the past year were captured. Primary outcomes were seven Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition-defined mood and anxiety disorders and eight personality disorders, which were measured with the Alcohol Use Disorder and Associated Disabilities Interview Schedule. RESULTS: Among pregnant women, 21.7% reported cigarette use and 12.4% met the criteria for nicotine dependence. Among pregnant women with cigarette use, 45.1% met criteria for at least one mental disorder, and among those with nicotine dependence, 57.5% met criteria for at least one other mental disorder. After adjusting for demographics and comorbidity, nicotine dependence during pregnancy significantly predicted any mental disorder (odds ratio [OR] 3.3, 95% confidence interval [CI] 2.1-5.1), any mood disorder (OR 2.5, 95% CI 1.5-4.0), major depression (OR 2.07, 95% CI 1.3 3.4), dysthymia (OR 6.2, 95% CI 2.9-13.1), and panic disorder (OR 3.1, 95% CI 1.6 6.1) in the past year. No significant associations were found between nondependent cigarette use and mental disorders. CONCLUSION: Our results suggest an association between mental disorders and nicotine dependence among pregnant women in the United States. This association has far-reaching implications for both the mental and physical health of women and potentially for their children. LEVEL OF EVIDENCE: III. PMID- 17400850 TI - Mifepristone and misoprostol administered simultaneously versus 24 hours apart for abortion: a randomized controlled trial. AB - OBJECTIVE: Mifepristone and oral misoprostol are typically used for medical abortion in women up to 49 days of gestation, with a 36- to 48-hour interval between the medications. Alternative routes of misoprostol administration allow for use beyond 49 days of gestation. We designed this randomized, noninferiority trial to compare the efficacy, adverse effects, and acceptability of misoprostol 800 mcg vaginally administered simultaneously with, or 24 hours after, mifepristone 200 mg orally for abortion in women up to 63 days of gestation. METHODS: The 1,128 participants swallowed mifepristone 200 mg and were then randomized to self-administer misoprostol intravaginally immediately in the office (group 1) or 24 hours later at home (group 2). Subjects returned for an evaluation, including transvaginal ultrasonography, 7+/-1 days after initiating treatment. Women who had not aborted were offered a second dose of misoprostol and returned for another evaluation in approximately 1 week. A phone contact was also attempted approximately 5 weeks after treatment. Treatment was considered a failure if a suction aspiration was performed for any indication. RESULTS: The complete abortion rate for group 1 (95.1%, 95% confidence interval [CI] 93.0 96.8%) was statistically noninferior to that for group 2 (96.9%, 95% CI 95.1 98.2%) (P=.003). The abortion rates between groups did not significantly differ by gestational age. Adverse effects were mostly similar, although nausea, diarrhea, and warmth or chills were significantly more common in group 1. CONCLUSION: Mifepristone 200 mg and misoprostol 800 mcg vaginally used simultaneously is as effective for abortion as compared with regimens using a 24 hour dosing interval. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00269568 LEVEL OF EVIDENCE: I. PMID- 17400851 TI - Mifepristone followed in 24 hours to 48 hours by misoprostol for late first trimester abortion. AB - OBJECTIVE: To investigate the efficacy of mifepristone and misoprostol for the termination of pregnancies in the late first trimester. METHODS: This was a prospective study of 321 women seeking termination of pregnancy with gestations from 64 days to 84 days (+/-3 days) by vaginal ultrasonography. Women were enrolled at three sites: University of Rochester Reproductive Health Program in Rochester, New York; Hung Vuong Hospital in Ho Chi Minh City, Vietnam; and K.E.M. Hospital in Pune, India. Eligible women received 800 mcg of misoprostol vaginally between 24 hours and 48 hours after administration of 200 mg mifepristone. Two additional doses of 400 mcg of misoprostol were administered either orally or vaginally as needed every 3 hours for a maximum of two additional doses (total 1,600 mcg). The primary study outcome measure was complete abortion without surgical intervention. RESULTS: Eighty-nine percent of women who completed the study successfully terminated their pregnancies. Most women were either satisfied (64.8%) or very satisfied (28.6%) with their experience. Ninety-four percent of women reported that they would recommend the procedure to a friend. Most women (90.4%) also agreed they would request a medical abortion if they required another abortion at this gestational age. CONCLUSION: Medical abortion is acceptable and effective in the late first trimester and offers women an acceptable alternative to surgical abortion. LEVEL OF EVIDENCE: II. PMID- 17400852 TI - Trauma and posttraumatic stress disorder in women with chronic pelvic pain. AB - OBJECTIVE: To examine the effect of abuse history, other major trauma, and posttraumatic stress disorder (PTSD) on medical symptoms and health-related daily functioning in women with chronic pelvic pain. METHODS: We administered a questionnaire to 713 consecutive women seen in a referral-based pelvic pain clinic. RESULTS: We found that 46.8% reported having either a sexual or physical abuse history. A total of 31.3% had a positive screen for PTSD. Using regression and path analysis, controlling for demographic variables, we found that a trauma history was associated with worse daily physical functioning due to poor health (P<.001), more medical symptoms (P<.001), more lifetime surgeries (P<.001), more days spent in bed (P<.001), and more dysfunction due to pain (P<.001). Furthermore, a positive screen for PTSD was highly related to most measures of poor health status (P<.001) and somewhat explained the trauma-related poor health status. CONCLUSION: The association of trauma with poor health may be due in part to the development of PTSD resulting from trauma. These findings demonstrate the importance of screening for trauma and PTSD in women with chronic pelvic pain. LEVEL OF EVIDENCE: II. PMID- 17400853 TI - Urinary incontinence in older community-dwelling women: the role of cognitive and physical function decline. AB - OBJECTIVES: To examine the association between cognitive decline, physical function decline, and urinary incontinence in older community-dwelling women. METHODS: This was an observational study of 6,361 community-dwelling women aged 65 years and older participating in the Study of Osteoporotic Fractures. Clinical frequency and functional disruptiveness of incontinence were assessed by self report questionnaires. Cognitive function was assessed at visits using the modified Mini-Mental State Examination, Trails B test, and Digit Symbol Substitution Test. Physical function was assessed by measuring walking speed over a 6-meter course and time needed to complete five chair stands. Women were considered to have recent, significant decline in cognitive or physical function if their cognitive or physical performance declined by greater than 1 standard deviation beyond the mean decline in the 6 years preceding assessment of incontinence. RESULTS: Women with recent physical function decline were more likely to report weekly incontinence (odds ratio [OR] 1.31, 95% confidence interval [CI] 1.09-1.56 for walking speed decline; OR 1.40, 95% CI 1.19-1.64 for chair stand decline), after adjusting for multiple characteristics. Women with recent cognitive decline were more likely to report incontinence that interfered with activities (OR 1.55, 95% CI 1.10-2.17 for modified Mini-Mental State Examination decline; OR 1.53, 95% CI 1.01-2.31 for Digit Symbol Substitution Test decline), after adjusting for multiple characteristics. CONCLUSION: Both cognitive and physical function decline are likely important contributors to incontinence in community-dwelling women aged 65 years and older. Although cognitive decline may not be associated with greater frequency of incontinence, women with cognitive decline may have more difficulty coping with incontinence symptoms. LEVEL OF EVIDENCE: II. PMID- 17400854 TI - Comparison of maternal and infant outcomes from primary cesarean delivery during the second compared with first stage of labor. AB - OBJECTIVE: To compare maternal and neonatal outcomes when primary cesarean delivery is performed in the second stage of labor compared with the first stage. METHODS: Between January 1, 1999, and December 31, 2000, a prospective observational study of primary cesarean deliveries was conducted at 13 university centers comprising the National Institute of Child Health and Human Development Maternal-Fetal Medicine Units Network. The primary outcomes of interest included a maternal composite (composed of at least one of the following: endometritis, intraoperative surgical complication, blood transfusion, or wound complication) and neonatal composite (which included at least one of the following: Apgar score of 3 or less at 5 minutes, neonatal death, neonatal intensive care unit admission, seizure, delivery room intubation in the absence of meconium, or fetal injury). RESULTS: A total of 11,981 cesarean deliveries were available for analysis: 9,265 were performed in the first stage and 2,716 in the second stage. Cesarean deliveries performed in the second stage were associated with longer operative times, epidural analgesia, chorioamnionitis, and higher birth weight (all P<.001). The maternal composite index was slightly increased in women undergoing cesarean delivery in the second stage of labor, primarily due to uterine atony, uterine incision extension, and incidental cystotomy. This difference was significant after multivariable analysis (odds ratio 1.21, 95% confidence interval 1.07-1.37). After multivariable analysis, the neonatal composite did not differ significantly between groups (odds ratio 0.96, 95% confidence interval 0.84-1.08). CONCLUSION: Cesarean delivery in the second stage of labor is associated with slightly increased maternal but not neonatal composite morbidity. LEVEL OF EVIDENCE: II. PMID- 17400855 TI - Urinary incontinence during pregnancy. AB - OBJECTIVE: To investigate incidence and prevalence of urinary incontinence during pregnancy and associated risk factors. METHODS: The data collection was conducted as part of the Norwegian Mother and Child Cohort Study at the Norwegian Institute of Public Health. We present questionnaire data about urinary incontinence obtained from 43,279 women (response rate 45%) by week 30. We report data on any incontinence, in addition to type, frequency, and amount of incontinence. Potential risk factors were investigated by logistic regression analyses. RESULTS: The prevalence of incontinence increased from 26% before pregnancy to 58% in week 30. The corresponding figures for nulliparous women were 15% and 48%, and for parous women 35% and 67%. The cumulative incidence was 46%. Stress urinary incontinence was the most common type of incontinence in week 30 of pregnancy, experienced by 31% of nulliparous and 42% of parous women. The majority of pregnant women had leakage less than once per week and droplets only, both before and during pregnancy. Parity was a strong and significant risk factor for incontinence in adjusted analyses both before pregnancy (odds ratio [OR] 2.5, 95% confidence interval [CI] 2.4-2.7 for primiparous and OR 3.3, 95% CI 3.1-3.5 for multiparous women) and during pregnancy (ORs 2.0, 95% CI 1.9-2.1 and 2.1, 95% CI 2.0-2.2, respectively). Age and body mass index were weaker, but still statistically significant, risk factors. CONCLUSION: The prevalence of urinary incontinence increases substantially during pregnancy. Incontinence both before and during pregnancy seems to be associated with parity, age, and body mass index. LEVEL OF EVIDENCE: II. PMID- 17400856 TI - Effect of war on the menstrual cycle. AB - OBJECTIVE: To study the effect of a short period of war on the menstrual cycles of exposed women. METHODS: Six months after a 16-day war, women in exposed villages aged 15-45 years were asked to complete a questionnaire relating to their menstrual history at the beginning, 3 months after, and 6 months after the war. A control group, not exposed to war, was also interviewed. The data collected were analyzed to estimate the effect of war on three groups of women: those who stayed in the war zone for 3-16 days (Group A), those who were displaced within 2 days to safer areas (Group B), and women not exposed to war or displacement (Group C-control). RESULTS: More than 35% of women in Group A and 10.5% in Group B had menstrual aberrations 3 months after the cessation of the war. These percentages were significantly different from each other and from that in Group C (2.6%). Six months after the war most women regained their regular menstrual cycles with the exception of 18.6% in Group A. CONCLUSION: We found a short period of war, acting like an acute stressful condition, resulted in menstrual abnormalities in 10-35% of women and is probably related to the duration of exposure to war. This might last beyond the war time and for more than one or two cycles. In most women the irregular cycles reversed without any medical intervention. LEVEL OF EVIDENCE: II. PMID- 17400857 TI - C-reactive protein concentrations and subsequent ovarian cancer risk. AB - OBJECTIVE: To estimate the association between prediagnostic levels of C-reactive protein (CRP), a marker of chronic systemic inflammation, and subsequent development of ovarian cancer. METHODS: A multicenter, nested, case-control study was conducted, including women who developed ovarian cancer (case patients) and women who were cancer-free (controls) from the following cohorts: CLUE ("Give us a CLUE to cancer and heart disease") cohorts of Washington County, Maryland, the Columbia, Missouri Serum Bank, and the Island of Guernsey Prospective Study, United Kingdom. A total of 167 incident invasive epithelial ovarian cancer cases were identified and each matched to an average of two controls on cohort, age, race, menopausal status, time since last menstrual period, current hormone use, date of recruitment, and time of day of blood draw. Baseline serum samples were assayed for CRP concentrations, and estimates of risk associated with CRP levels were assessed using conditional logistic regression. RESULTS: Ovarian cancer risk was positively associated with increasing CRP concentrations. The risk of developing ovarian cancer among women in the highest third of the distribution of CRP compared with those in the lowest third was 1.72 (95% confidence interval 1.06-2.77), with evidence of an increasing risk with increasing concentration of CRP (P trend=0.02). Similar associations were observed using established clinical CRP cutpoints for heart disease risk (odds ratio 2.03, 95% confidence interval 1.20-3.47 for 3-10 mg/L compared with less than 1 mg/L, P trend=.008). If this association is causal, roughly 23% of ovarian cancer cases are attributed to chronic inflammation as indicated by elevated CRP concentrations. CONCLUSION: Higher circulating CRP concentrations in women who subsequently developed ovarian cancer support the hypothesized role of chronic inflammation in ovarian carcinogenesis. LEVEL OF EVIDENCE: II. PMID- 17400858 TI - Surgical treatments for vulvar and vaginal dysplasia: a randomized controlled trial. AB - OBJECTIVE: To compare pain, adverse effects and recurrence of dysplasia in patients with vaginal intraepithelial neoplasia or vulvar intraepithelial neoplasia prospectively treated by carbon dioxide laser or ultrasonic surgical aspiration. METHODS: Patients were randomly assigned to receive treatment by laser or ultrasonic surgical aspiration from 2000-2005. Preoperative biopsy was done to confirm presence of dysplasia. Patients completed a visual analog scale regarding pain and were evaluated at 2-4 weeks to assess scarring, wound healing, and adverse effects. Patients returned every 3 months for 1 year for pelvic examination and cytology to assess recurrence. Follow-up colposcopy and biopsy were used at the discretion of the treating physician. Student t test, chi2, analysis of variance and multiple logistic regression were used for analysis. RESULTS: One hundred ten patients were randomly assigned. Ninety-six (87.3%) patients completed 1 year follow-up. Mean age of patients was 48.5 years. Mean visual analog scale score was significantly lower in patients treated by ultrasonic surgical aspiration (20.7 compared with 35.1; P=.032). For patients with vulvar lesions, there was less scarring with ultrasonic surgical aspiration (P<.01). Recurrence overall was 25% and was similar for ultrasonic surgical aspiration compared with laser (relative risk 0.96, 95% confidence interval 0.64 1.50, number needed to treat 95.6). Recurrence was associated with younger age (P<.01). CONCLUSION: Patients treated with ultrasonic surgical aspiration for vulvar and vaginal dysplasia reported less postoperative pain. Vulvar scarring was more common in patients treated by the laser. There was no difference in recurrence of dysplasia during a 1-year follow-up period between the two surgical modalities. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, www.clinicaltrials.gov, NCT00394758 LEVEL OF EVIDENCE: I. PMID- 17400859 TI - Predictors of physician career satisfaction, work-life balance, and burnout. AB - OBJECTIVE: To explore factors associated with physician career satisfaction, work life balance, and burnout focusing on differences across age, gender, and specialty. METHODS: A cross-sectional, mailed, self-administered survey was sent to a national sample of 2,000 randomly-selected physicians, stratified by specialty, age, and gender (response rate 48%). Main outcome measures included career satisfaction, burnout, and work-life balance. Scales ranged from 1 to 100. RESULTS: Both women and men report being highly satisfied with their careers (79% compared with 76%, P<.01), having moderate levels of satisfaction with work-life balance (48% compared with 49%, P=.24), and having moderate levels of emotional resilience (51% compared with 53%, P=.09). Measures of burnout strongly predicted career satisfaction (standardized beta 0.36-0.60, P<.001). The strongest predictor of work-life balance and burnout was having some control over schedule and hours worked (standardized beta 0.28, P<.001, and 0.20-0.32, P<.001, respectively). Physician gender, age, and specialty were not strong independent predictors of career satisfaction, work-life balance, or burnout. CONCLUSION: This national physician survey suggests that physicians can struggle with work life balance yet remain highly satisfied with their career. Burnout is an important predictor of career satisfaction, and control over schedule and work hours are the most important predictors of work-life balance and burnout. LEVEL OF EVIDENCE: II. PMID- 17400860 TI - Imitators of severe preeclampsia. AB - There are several obstetric, medical, and surgical disorders that share many of the clinical and laboratory findings of patients with severe preeclampsia hemolysis, elevated liver enzymes, and low platelets syndrome. Imitators of severe preeclampsia-hemolysis, elevated liver enzymes, and low platelets syndrome are life-threatening emergencies that can develop during pregnancy or in the postpartum period. These conditions are associated with high maternal mortality, and survivors may face long-term sequelae. Perinatal mortality and morbidity also remain high in many of these conditions. The pathophysiologic abnormalities in many of these disorders include thrombotic microangiopathy, thrombocytopenia, and hemolytic anemia. Some of these disorders include acute fatty liver of pregnancy, thrombotic thrombocytopenic purpura, hemolytic uremic syndrome, and acute exacerbation of systemic lupus erythematosus. Because of the rarity of these conditions during pregnancy and postpartum, the available literature includes only case reports and case series describing these syndromes. Consequently, there are no systematic reviews or randomized trials on these subjects. Differential diagnosis may be difficult due to the overlap of several clinical and laboratory findings of these syndromes. It is important that the clinician make the accurate diagnosis when possible because the management and complications from these syndromes may be different. For example, severe preeclampsia and acute fatty liver of pregnancy are treated by delivery, whereas it is possible to continue pregnancy in those with thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and exacerbation of systemic lupus erythematosus. This review focuses on diagnosis, management, and counseling of women who develop these syndromes based on results of recent studies. PMID- 17400861 TI - Infertility, assisted reproductive technology, and adverse pregnancy outcomes: executive summary of a National Institute of Child Health and Human Development workshop. AB - The National Institute of Child Health and Human Development held a workshop on September 12-13, 2005, to summarize the risks for adverse pregnancy outcomes after assisted reproductive technology (ART), develop an approach to counseling couples regarding these risks, and establish a research agenda. Although the majority of ART children are normal, there are concerns about the increased risk for adverse pregnancy outcomes. More than 30% of ART pregnancies are twins or higher-order multiple gestations (triplets or greater) and more than one half of all ART neonates are the products of multifetal gestations, with an attendant increase in prematurity complications. Assisted reproductive technology singleton pregnancies also demonstrate increased rates of perinatal complications-small for gestational age infants, preterm delivery, and perinatal mortality-as well as maternal complications, such as preeclampsia, gestational diabetes, placenta previa, placental abruption, and cesarean delivery. Although it is not possible to separate ART-related risks from those secondary to the underlying reproductive pathology, the overall increased frequency of obstetric complications, including preterm birth and small for gestational age neonates, should be discussed with the couple. Significant gaps in knowledge were identified, and the basic science and clinical and epidemiologic research required to address these gaps is outlined. PMID- 17400862 TI - Risks, values, and decision making surrounding pregnancy. AB - Assessing, communicating, and managing risk are among the most challenging tasks in the practice of medicine and are particularly difficult in the context of pregnancy. We analyze common scenarios in medical decision making around pregnancy, from reproductive health policy and clinical care to research protections. We describe three tendencies in these scenarios: 1) to consider the probabilities of undesirable outcomes alone, in isolation from women's values and social contexts, as determinative of individual clinical decisions and health policy; 2) to regard any risk to the fetus, including incremental risks that would in other contexts be regarded as acceptable, as trumping considerations that may be substantially more important to the wellbeing of the pregnant woman; and 3) to focus on the risks associated with undertaking medical interventions during pregnancy to the exclusion of demonstrable risks to both woman and fetus of failing to intervene. These tendencies in the perception, communication, and management of risk can lead to care that is neither evidence-based nor patient centered, often to the detriment of both women and infants. PMID- 17400863 TI - Constipation: will the bowel recipe do the trick? PMID- 17400865 TI - Intravenous immunoglobulin (IVIG). PMID- 17400864 TI - A new progestin implant (implanon) for long-term contraception. PMID- 17400867 TI - IN brief: clarification: hand hygiene and CDAD. PMID- 17400866 TI - Treatment of Clostridium difficile--associated disease (CDAD). PMID- 17400868 TI - Risk of urinary incontinence after childbirth: a 10-year prospective cohort study. PMID- 17400869 TI - U.S. Food and drug administration analysis of strokes associated with raloxifene. PMID- 17400870 TI - Association of lymphadenectomy and survival in stage I ovarian cancer patients. PMID- 17400871 TI - Chronic abdominal pain after laparoscopic sterilization clip placement. PMID- 17400872 TI - ACOG Practice Bulletin No. 80: premature rupture of membranes. Clinical management guidelines for obstetrician-gynecologists. AB - Preterm delivery occurs in approximately 12% of all births in the United States and is a major factor contributing to perinatal morbidity and mortality (1, 2). Despite extensive research in this area, the rate of preterm birth has increased by 38% since 1981 (3). Premature rupture of membranes (PROM) is a complication in approximately one third of preterm births. It typically is associated with brief latency between membrane rupture and delivery, increased potential for perinatal infection, and in utero umbilical cord compression. Because of this, both PROM at and before term can lead to significant perinatal morbidity and mortality. There is some controversy over the optimal approaches to clinical assessment and treatment of women with term and preterm PROM. Management hinges on knowledge of gestational age and evaluation of the relative risks of preterm birth versus intrauterine infection, abruptio placentae, and cord accident that could occur with expectant management. The purpose of this document is to review the current understanding of this condition and to provide management guidelines that have been validated by appropriately conducted outcome-based research. Additional guidelines on the basis of consensus and expert opinion also are presented. PMID- 17400873 TI - ACOG Committee Opinion No. 363: patient testing: ethical issues in selection and counseling. AB - Recommendations to patients about testing should be based on current medical knowledge, a concern for the patient's best interests, and mutual consultation. In addition to establishing a diagnosis, testing provides opportunities to educate, inform, and advise. The ethical principles of respect for autonomy (patient choice) and beneficence (concern for the patient's best interests) should guide the testing, counseling, and reporting process. Clear and ample communication fosters trust, facilitates access to services, and improves the quality of medical care. PMID- 17400874 TI - Antimicrobial peptides in the real world: implications for cystic fibrosis. PMID- 17400875 TI - Who's (still) afraid of talc? PMID- 17400876 TI - Right ventricular ejection fraction and NT-proBNP are both indicators of wall stress in pulmonary hypertension. PMID- 17400877 TI - Travel to high altitude with pre-existing lung disease. AB - The pathophysiology of high-altitude illnesses has been well studied in normal individuals, but little is known about the risks of high-altitude travel in patients with pre-existing lung disease. Although it would seem self-evident that any patient with lung disease might not do well at high altitude, the type and severity of disease will determine the likelihood of difficulty in a high altitude environment. The present review examines whether these individuals are at risk of developing one of the main forms of acute or chronic high-altitude illness and whether the underlying lung disease itself will get worse at high elevations. Several groups of pulmonary disorders are considered, including obstructive, restrictive, vascular, control of ventilation, pleural and neuromuscular diseases. Attempts will be made to classify the risks faced by each of these groups at high altitude and to provide recommendations regarding evaluation prior to high-altitude travel, advice for or against taking such excursions, and effective prophylactic measures. PMID- 17400878 TI - Local genetic and environmental factors in asthma disease pathogenesis: chronicity and persistence mechanisms. AB - While asthma is an inflammatory disorder of the airways usually associated with atopy, an important additional component is involvement of the epithelium and underlying mesenchyme acting as a trophic unit (EMTU). In addition to allergens, a wide range of environmental factors interact with the EMTU, such as virus infections, environmental tobacco smoke and pollutants, to initiate tissue damage and aberrant repair responses that are translated into remodelling of the airways. While candidate gene association studies have revealed polymorphic variants that influence asthmatic inflammation, positional cloning of previously unknown genes is identifying a high proportion of novel genes in the EMTU. Dipeptidyl peptidase (DPP) 10 and disintegrin and metalloproteinase (ADAM)33 are newly identified genes strongly associated with asthma that are preferentially expressed in the airway epithelium and underlying mesenchyme, respectively. Also of increasing importance is the recognition that genes associated with asthma and atopy have important interactions with the environment through epigenetic mechanisms that influence their expression. This type of research will not only identify biomarkers of different types of asthma across the full range of phenotypic expression, but will also identify novel therapeutic targets that could influence the natural history of the heterogenes lung disease. PMID- 17400879 TI - Master switches of T-cell activation and differentiation. AB - T-cells play a central role in allergic airway diseases such as bronchial asthma. The imbalance between allergen-specific pro-inflammatory and pro-allergic T-cell responses on one hand and regulatory or suppressive T-cell responses on the other may best explain the development of unwanted immune responses against environmental allergens, which lead to immunoglobulin E production and airway inflammation. A key role in the fine tuning of any T-cell response is provided by the engagement of so-called co-stimulatory molecules that are required for the full activation of T-cells and the recognition of antigens via the antigen specific T-cell receptor. Many of these co-stimulatory molecules have been identified only recently, leading to a fundamental change in the overall understanding of T-cell regulation. Due to their pivotal impact on T-cell differentiation and control, co-stimulatory molecules are promising targets for therapeutic intervention in T-cell-regulated or -mediated immune disorders, including allergic diseases and asthma. In the present article, an attempt is made to summarise the current knowledge on the basic concept of co-stimulation, the presently known co-stimulatory molecules and their various functions on T cell activation or suppression. The mini-series will be completed by two more articles describing the recent experimental studies and preliminary clinical findings regarding the role of co-stimulatory molecules in allergic disorders and bronchial asthma, and a discussion regarding the feasibility of co-stimulatory molecules as potential targets for the treatment of allergic airway disease. Although it is too early for any clinical implication or utilisation at this moment, the authors are convinced that a better understanding of co-stimulation in the context of allergic asthma will finally provide novel and promising approaches for treatment and prevention. PMID- 17400880 TI - Pulmonary arteriovenous fistula in a patient with renal cell carcinoma. AB - Arteriovenous (AV) fistulas are common in renal cell carcinoma. These are usually small and asymptomatic. Case reports have previously described symptomatic fistulas in primary renal cell tumour or, less commonly, bone metastases. The current study describes a patient with lung metastases from renal cell carcinoma who developed a symptomatic pulmonary AV fistula. The patient presented with debilitating progressive shortness of breath, which resolved after coiling of the pulmonary AV fistula. Supporting radiographs include a unique ventilation perfusion finding of perfusion tracer in the kidney that is diagnostic of a pulmonary shunt. This is the first report of a pulmonary fistula from renal cell carcinoma. This rare complication of renal cell carcinoma reflects its diverse clinical presentations and unique tumour biology. PMID- 17400881 TI - Angiotensin-converting enzyme genotype and C-reactive protein in patients with COPD. PMID- 17400882 TI - Relapsing necrotising sarcoid granulomatosis in a young patient. PMID- 17400883 TI - Diffuse panbronchiolitis: East meets West. PMID- 17400884 TI - Exposure to second-hand smoke: a population-based survey in Spain. PMID- 17400885 TI - Effect of spirometry on COPD management in primary care: where are the studies that we really need? PMID- 17400886 TI - Evidence-based medicines for children: ethical aspects. PMID- 17400887 TI - Neuroinflammation and behavioral abnormalities after neonatal terbutaline treatment in rats: implications for autism. AB - Autism is a neurodevelopmental disorder presenting before 3 years of age with deficits in communication and social skills and repetitive behaviors. In addition to genetic influences, recent studies suggest that prenatal drug or chemical exposures are risk factors for autism. Terbutaline, a beta2-adrenoceptor agonist used to arrest preterm labor, has been associated with increased concordance for autism in dizygotic twins. We studied the effects of terbutaline on microglial activation in different brain regions and behavioral outcomes in developing rats. Newborn rats were given terbutaline (10 mg/kg) daily on postnatal days (PN) 2 to 5 or PN 11 to 14 and examined 24 h after the last dose and at PN 30. Immunohistochemical studies showed that administration of terbutaline on PN 2 to 5 produced a robust increase in microglial activation on PN 30 in the cerebral cortex, as well as in cerebellar and cerebrocortical white matter. None of these effects occurred in animals given terbutaline on PN 11 to 14. In behavioral tests, animals treated with terbutaline on PN 2 to 5 showed consistent patterns of hyper-reactivity to novelty and aversive stimuli when assessed in a novel open field, as well as in the acoustic startle response test. Our findings indicate that beta2-adrenoceptor overstimulation during an early critical period results in microglial activation associated with innate neuroinflammatory pathways and behavioral abnormalities, similar to those described in autism. This study provides a useful animal model for understanding the neuropathological processes underlying autism spectrum disorders. PMID- 17400888 TI - Selectivity of delta- and kappa-opioid ligands depends on the route of central administration in mice. AB - The existence of heterodimeric opioid receptors has introduced greater complexity to the in vivo characterization of pharmacological selectivity of agonists by antagonists. Because of the possibility of cooperativity between receptors organized as heterodimers, it is conceivable that selective antagonists may antagonize an agonist bound to a neighboring, allosterically coupled receptor. As a consequence, the in vivo selectivity of an opioid antagonist may depend on the organizational state of receptors that mediate analgesia. In this regard, phenotypic delta- and kappa-opioid receptors have been proposed to arise from different organizational states that include oligomeric delta-kappa heterodimers and homomeric delta and kappa receptors. In view of the evidence for analgesia mediated by delta-kappa heterodimers in the spinal cord, but not the brain, we have investigated the selectivity of pharmacologically selective delta and kappa antagonists in mice by both i.t. and i.c.v. routes of administration to evaluate changes in selectivity. Using pharmacologically selective delta (benzylidenenaltrexone, naltrindole, and naltriben) and kappa (norbinaltorphimine) antagonists versus delta ([D-Pen(2),D-Pen(5)]-enkephalin and deltorphin II) and kappa [3,4-dichloro-N-methyl-N-[(1R,2R)-2-(1 pyrrolidinyl)cyclohexyl]-benzeneacetamide (U50488) and bremazocine] agonists, the delta-1/delta-2 selectivity ratios were found to be dependent on the route of administration (i.t. versus i.c.v.). The data from different routes of administration suggest that differences in molecular recognition between spinal delta-kappa heterodimers and supraspinal homomeric delta and kappa receptors may contribute to the divergent selectivity ratios of selective antagonists. In view of the observed tissue-dependent selectivity, we suggest that multiple opioid antagonists be employed routinely in establishing agonist selectivity in vivo. PMID- 17400889 TI - Cisplatin-induced acute renal failure is associated with an increase in the cytokines interleukin (IL)-1beta, IL-18, IL-6, and neutrophil infiltration in the kidney. AB - We have demonstrated that caspase-1-deficient (caspase-1(-/-)) mice are functionally and histologically protected against cisplatin-induced acute renal failure (ARF). Caspase-1 exerts proinflammatory effects via the cytokines interleukin (IL)-1beta, IL-18, IL-6, and neutrophil recruitment. We sought to determine the role of the cytokines IL-1beta, IL-18, and IL-6 and neutrophil recruitment in cisplatin-induced ARF. We first examined IL-1beta; renal IL-1beta increased nearly 2-fold in cisplatin-induced ARF and was reduced in the caspase 1(-/-) mice. However, inhibition with IL-1 receptor antagonist (IL-1Ra) did not attenuate cisplatin-induced ARF. Renal IL-18 increased 2.5-fold; however, methods to inhibit IL-18 using IL-18 antiserum and transgenic mice that overproduce IL-18 binding protein (a natural inhibitor of IL-18) did not protect. Renal IL-6 increased 3-fold; however, IL-6-deficient (IL-6(-/-)) mice still developed cisplatin-induced ARF. We next examined neutrophils; blood neutrophils increased dramatically after cisplatin injection; however, prevention of peripheral neutrophilia and renal neutrophil infiltration with the neutrophil-depleting antibody RB6-8C5 did not protect against cisplatin-induced ARF. In summary, our data demonstrated that cisplatin-induced ARF is associated with increases in the cytokines IL-1beta, IL-18, and IL-6 and neutrophil infiltration in the kidney. However, inhibition of IL-1beta, IL-18, and IL-6 or neutrophil infiltration in the kidney is not sufficient to prevent cisplatin-induced ARF. PMID- 17400890 TI - Apoptosis in microencapsulated juvenile rabbit chondrocytes induced by ofloxacin: role played by beta(1)-integrin receptor. AB - Quinolone(s) (QNs) is widely used in infection therapy due to its good antimicrobial characteristics. However, QNs-induced arthropathy of immature animals has led to restrictions on the therapeutic use of these antimicrobial agents. The exact mechanism(s) of QNs-induced chondrotoxicity remain unknown. In the present study, we investigated the possible mechanism of ofloxacin (one typical QNs)-induced injuries of chondrocytes. Juvenile rabbit joint chondrocytes cultured in alginate microspheres were incubated with ofloxacin at concentrations of 0, 2, 5, 10, 20, and 40 microg/ml for up to 96 h. Concentration of 10 microg/ml ofloxacin induced apoptosis of chondrocyte with visible apoptotic signs, including degradation of poly(ADP-ribose) polymerase, caspase-3 activation, and DNA ladder formation. Furthermore, extracellular signal-regulated kinase 1/2 (phospho-ERK1/2) and growth factor receptor-bound protein 2 (Grb2) were significantly reduced, and similar changes were also observed in the beta(1) integrin receptor as assessed by immunoblotting. However, the mRNA level of beta(1)-integrin obtained from reverse transcription-polymerase chain reaction remained unchanged. Results of beta(1)-integrin immunoprecipitation have also shown that beta(1)-integrin did not interact with activated intracellular signaling proteins. In addition, ofloxacin did not induce apoptosis and decrease beta(1)-integrin expression in chondrocytes supplemented with Mg(2+), and the ofloxacin-induced apoptosis was caspase-8-dependent, inhibition of which did not affect the expression mode of phospho-ERK1/2 and beta(1)-integrin. Our results demonstrate that ofloxacin affects beta(1)-integrin receptor functions and the ERK mitogen-activated protein kinase signaling pathway, causing caspase-8 dependent apoptosis after exposure of 48 h. PMID- 17400891 TI - Chitosan, the deacetylated form of chitin, is necessary for cell wall integrity in Cryptococcus neoformans. AB - Cryptococcus neoformans is an opportunistic fungal pathogen that causes cryptococcal meningoencephalitis, particularly in immunocompromised patients. The fungal cell wall is an excellent target for antifungal therapies as it is an essential organelle that provides cell structure and integrity, it is needed for the localization or attachment of known virulence factors, including the polysaccharide capsule, melanin, and phospholipase, and it is critical for host pathogen interactions. In C. neoformans, chitosan produced by the enzymatic removal of acetyl groups from nascent chitin polymers has been implicated as an important component of the vegetative cell wall. In this study, we identify four putative chitin/polysaccharide deacetylases in C. neoformans. We have demonstrated that three of these deacetylases, Cda1, Cda2, and Cda3, can account for all of the chitosan produced during vegetative growth in culture, but the function for one, Fpd1, remains undetermined. The data suggest a model for chitosan production in vegetatively growing C. neoformans where the three chitin deacetylases convert chitin generated by the chitin synthase Chs3 into chitosan. Utilizing a collection of chitin/polysaccharide deacetylase deletion strains, we determined that during vegetative growth, chitosan helps to maintain cell integrity and aids in bud separation. Additionally, chitosan is necessary for maintaining normal capsule width and the lack of chitosan results in a "leaky melanin" phenotype. Our analysis indicates that chitin deacetylases and the chitosan made by them may prove to be excellent antifungal targets. PMID- 17400892 TI - Elaboration of B gene function to include the identity of novel floral organs in the lower eudicot Aquilegia. AB - The basal eudicot Aquilegia (columbine) has an unusual floral structure that includes two morphologically distinct whorls of petaloid organs and a clearly differentiated fifth organ type, the staminodium. In this study, we have sought to determine how Aquilegia homologs of the B class genes APETALA3 (AP3) and PISTILLATA (PI) contribute to these novel forms of organ identity. Detailed expression analyses of the three AP3 paralogs and one PI homolog in wild-type and floral homeotic mutant lines reveal complex patterns that suggest that canonical B class function has been elaborated in Aquilegia. Yeast two-hybrid studies demonstrate that the protein products of Aquilegia's AP3 and PI homologs can form heterodimers, much like what has been observed for their core eudicot homologs. Downregulation of AqvPI using virus-induced gene silencing indicates that in addition to petal and stamen identity, this locus is essential to staminodial identity but may not control the identity of the petaloid sepals. Our findings show that preexisting floral organ identity programs can be partitioned and modified to produce additional organ types. In addition, they indicate that some types of petaloid organs are not entirely dependent on AP3/PI homologs for their identity. PMID- 17400893 TI - Genome-wide analysis of the RNA-DEPENDENT RNA POLYMERASE6/DICER-LIKE4 pathway in Arabidopsis reveals dependency on miRNA- and tasiRNA-directed targeting. AB - Posttranscriptional RNA silencing of many endogenous transcripts, viruses, and transgenes involves the RNA-DEPENDENT RNA POLYMERASE6/DICER-LIKE4 (RDR6/DCL4) dependent short interfering RNA (siRNA) biogenesis pathway. Arabidopsis thaliana contains several families of trans-acting siRNAs (tasiRNAs) that form in 21 nucleotide phased arrays through the RDR6/DCL4-dependent pathway and that negatively regulate target transcripts. Using deep sequencing technology and computational approaches, the phasing patterns of known tasiRNAs and tasiRNA-like loci from across the Arabidopsis genome were analyzed in wild-type plants and silencing-defective mutants. Several gene transcripts were found to be routed through the RDR6/DCL4-dependent pathway after initial targeting by one or multiple miRNAs or tasiRNAs, the most conspicuous example of which was an expanding clade of genes encoding pentatricopeptide repeat (PPR) proteins. Interestingly, phylogenetic analysis using Populus trichocarpa revealed evidence for small RNA-mediated regulatory mechanisms within a similarly expanded group of PPR genes. We suggest that posttranscriptional silencing mechanisms operate on an evolutionary scale to buffer the effects of rapidly expanding gene families. PMID- 17400894 TI - Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. AB - Mitogen-activated protein kinase (MAPK) signaling plays a central role in transducing extracellular stimuli into intracellular responses, but its role in mediating plant responses to herbivore attack remains largely unexplored. When Manduca sexta larvae attack their host plant, Nicotiana attenuata, the plant's wound response is reconfigured at transcriptional, phytohormonal, and defensive levels due to the introduction of oral secretions (OS) into wounds during feeding. We show that OS dramatically amplify wound-induced MAPK activity and that fatty acid-amino acid conjugates in M. sexta OS are the elicitors. Virus induced gene silencing of salicylic acid-induced protein kinase (SIPK) and wound induced protein kinase revealed their importance in mediating wound and OS elicited hormonal responses and transcriptional regulation of defense-related genes. We found that after applying OS to wounds created in one portion of a leaf, SIPK is activated in both wounded and specific unwounded regions of the leaf but not in phylotactically connected adjacent leaves. We propose that M. sexta attack elicits a mobile signal that travels to nonwounded regions of the attacked leaf where it activates MAPK signaling and, thus, downstream responses; subsequently, a different signal is transported by the vascular system to systemic leaves to initiate defense responses without activating MAPKs in systemic leaves. PMID- 17400895 TI - Calcium-dependent protein kinases regulate the production of reactive oxygen species by potato NADPH oxidase. AB - Reactive oxygen species (ROS) are implicated in plant innate immunity. NADPH oxidase (RBOH; for Respiratory Burst Oxidase Homolog) plays a central role in the oxidative burst, and EF-hand motifs in the N terminus of this protein suggest possible regulation by Ca(2+). However, regulatory mechanisms are largely unknown. We identified Ser-82 and Ser-97 in the N terminus of potato (Solanum tuberosum) St RBOHB as potential phosphorylation sites. An anti-phosphopeptide antibody (pSer82) indicated that Ser-82 was phosphorylated by pathogen signals in planta. We cloned two potato calcium-dependent protein kinases, St CDPK4 and St CDPK5, and mass spectrometry analyses showed that these CDPKs phosphorylated only Ser-82 and Ser-97 in the N terminus of St RBOHB in a calcium-dependent manner. Ectopic expression of the constitutively active mutant of St CDPK5, St CDPK5VK, provoked ROS production in Nicotiana benthamiana leaves. The CDPK-mediated ROS production was disrupted by knockdown of Nb RBOHB in N. benthamiana. The loss of function was complemented by heterologous expression of wild-type potato St RBOHB but not by a mutant (S82A/S97A). Furthermore, the heterologous expression of St CDPK5VK phosphorylated Ser-82 of St RBOHB in N. benthamiana. These results suggest that St CDPK5 induces the phosphorylation of St RBOHB and regulates the oxidative burst. PMID- 17400896 TI - Arabidopsis phage-type RNA polymerases: accurate in vitro transcription of organellar genes. AB - The T7 bacteriophage RNA polymerase (RNAP) performs all steps of transcription, including promoter recognition, initiation, and elongation as a single polypeptide enzyme. Arabidopsis thaliana possesses three nuclear-encoded T7 phage type RNAPs that localize to mitochondria (RpoTm), plastids (RpoTp), or presumably both organelles (RpoTmp). Their specific functions are as yet unresolved. We have established an in vitro transcription system to examine the abilities of the three Arabidopsis phage-type RNAPs to synthesize RNA and to recognize organellar promoters. All three RpoT genes were shown to encode transcriptionally active RNAPs. RpoTmp displayed no significant promoter specificity, whereas RpoTm and RpoTp were able to accurately initiate transcription from overlapping subsets of mitochondrial and plastidial promoters without the aid of protein cofactors. Our study strongly suggests RpoTm to be the enzyme that transcribes most, if not all, mitochondrial genes in Arabidopsis. Intrinsic promoter specificity, a feature that RpoTm and RpoTp share with the T7 RNAP, appears to have been conserved over the long period of evolution of nuclear-encoded mitochondrial and plastidial RNAPs. Selective promoter recognition by the Arabidopsis phage-type RNAPs in vitro implies that auxiliary factors are required for efficient initiation of transcription in vivo. PMID- 17400897 TI - Rice UDP-glucose pyrophosphorylase1 is essential for pollen callose deposition and its cosuppression results in a new type of thermosensitive genic male sterility. AB - UDP-glucose pyrophosphorylase (UGPase) catalyzes the reversible production of glucose-1-phosphate and UTP to UDP-glucose and pyrophosphate. The rice (Oryza sativa) genome contains two homologous UGPase genes, Ugp1 and Ugp2. We report a functional characterization of rice Ugp1, which is expressed throughout the plant, with highest expression in florets, especially in pollen during anther development. Ugp1 silencing by RNA interference or cosuppression results in male sterility. Expressing a double-stranded RNA interference construct in Ugp1-RI plants resulted in complete suppression of both Ugp1 and Ugp2, together with various pleiotropic developmental abnormalities, suggesting that UGPase plays critical roles in plant growth and development. More importantly, Ugp1 cosuppressing plants contained unprocessed intron-containing primary transcripts derived from transcription of the overexpression construct. These aberrant transcripts undergo temperature-sensitive splicing in florets, leading to a novel thermosensitive genic male sterility. Pollen mother cells (PMCs) of Ugp1-silenced plants appeared normal before meiosis, but during meiosis, normal callose deposition was disrupted. Consequently, the PMCs began to degenerate at the early meiosis stage, eventually resulting in complete pollen collapse. In addition, the degeneration of the tapetum and middle layer was inhibited. These results demonstrate that rice Ugp1 is required for callose deposition during PMC meiosis and bridges the apoplastic unloading pathway and pollen development. PMID- 17400899 TI - Will bluetongue come on the wind to the United Kingdom in 2007? AB - In 2006, over 2000 cases of bluetongue were recorded in northern Europe. The disease, which has been more typically associated with Mediterranean areas, is believed to have become established hundreds of kilometres to the north of its traditional area, probably as a consequence of the hottest summer/autumn period since records began. In this special article, John Gloster and colleagues describe the meteorological conditions surrounding the 2006 outbreak, and investigate the possibility of bluetongue virus (BTV) spreading on the wind to the UK in 2007. For this to happen there would need to be a source of windborne virus, together with a susceptible population of ruminants in the vicinity of the coast. Evidence from outbreaks in the Mediterranean Basin suggests that long distance transport of BTV-infected vectors has already occurred, at least in that region. The overall likelihood of this occurring in northern Europe depends critically on whether the virus overwinters on the near continent; this will not be known until around May 2007. The 2006 outbreak has highlighted the importance of understanding the impact of climate change on animal disease. PMID- 17400898 TI - A mutant of the Arabidopsis phosphate transporter PHT1;1 displays enhanced arsenic accumulation. AB - The exceptional toxicity of arsenate [As(V)] is derived from its close chemical similarity to phosphate (Pi), which allows the metalloid to be easily incorporated into plant cells through the high-affinity Pi transport system. In this study, we identified an As(V)-tolerant mutant of Arabidopsis thaliana named pht1;1-3, which harbors a semidominant allele coding for the high-affinity Pi transporter PHT1;1. pht1;1-3 displays a slow rate of As(V) uptake that ultimately enables the mutant to accumulate double the arsenic found in wild-type plants. Overexpression of the mutant protein in wild-type plants provokes phenotypic effects similar to pht1;1-3 with regard to As(V) uptake and accumulation. In addition, gene expression analysis of wild-type and mutant plants revealed that, in Arabidopsis, As(V) represses the activation of genes specifically involved in Pi uptake, while inducing others transcriptionally regulated by As(V), suggesting that converse signaling pathways are involved in plant responses to As(V) and low Pi availability. Furthermore, the repression effect of As(V) on Pi starvation responses may reflect a regulatory mechanism to protect plants from the extreme toxicity of arsenic. PMID- 17400900 TI - Long-term treatment with carprofen of 805 dogs with osteoarthritis. AB - The pain-relieving effect of carprofen and tolerance to the drug were investigated in 805 dogs that were lame as a result of osteoarthritis. The dogs were of different breeds, ages and bodyweights and of both sexes, and were selected from 51 veterinary clinics. Each dog was treated orally by its owner with 4 mg/kg carprofen for 84 consecutive days. Twenty-four dogs were removed from the study because of side effects, and 55 left the study for reasons unrelated to the treatment. The condition of the dogs and the benefit of the treatment were evaluated by the veterinary surgeons and the owners after 14 days, and at the end of the period of treatment, when 194 of the dogs (26.7 per cent) were no longer lame, and 357 (49.2 per cent) had improved. The period for which the dogs had been lame before entering the study significantly (P<0.01) affected the results and the rate of improvement. Too much exercise during the 84 days of treatment caused some dogs to relapse. PMID- 17400901 TI - Transient adverse effects of an attenuated bluetongue virus vaccine on the quality of ram semen. AB - This study investigated the effects of the vaccination of rams with a serotype 2 bluetongue virus vaccine on the quality of their semen. One group of 23 rams was vaccinated on days 0 and 47, and 23 rams were left unvaccinated. Samples of blood, serum and semen were collected regularly in order to detect the virus genome, and to compare the quality of the semen from the vaccinated and unvaccinated rams. Segment 10 of the genome of the vaccine strain was detected in the blood of the vaccinated animals by reverse transcriptase-PCR (RT-PCR) on days 7, 13 and 19 after the first vaccination, but no virus was isolated from the RT PCR-positive blood or from any of the semen samples from the vaccinated animals. There was a significant decrease in the concentration and motility of the spermatozoa and an increase in the proportion of abnormal and dead spermatozoa after the first vaccination; however, after the second vaccination only smaller, non-significant changes were observed. On day 69, the quality of the semen of the vaccinated animals was not significantly different from that of the controls. PMID- 17400902 TI - Field technique for the resection of the distal interphalangeal joint and proximal resection of the deep digital flexor tendon in cows. AB - A modified technique for the resection of the distal interphalangeal joint and the proximal resection of the deep digital flexor tendon in cows is described. Septic arthritis of the joint was diagnosed in eight Holstein cows and treated in the field. Four of the cows were diagnosed with ascending tendonitis during the resection of the joint and their tendons were also resected. All the animals remained moderately to severely lame for two weeks postoperatively but quickly recovered and were sound within five months. Eight months after the last surgery only one cow had been culled, 321 days after its surgery, for reproductive failure. The other seven cows had survived for a mean period of 308 days, with a range from 235 to 392 days. PMID- 17400903 TI - Apparent ivermectin resistance of Parascaris equorum in foals in Denmark. PMID- 17400904 TI - Ocular and cardiac malformations associated with maternal hypovitaminosis A in cattle. PMID- 17400905 TI - Measurement of distal motor latency of the suprascapular nerve in dogs. PMID- 17400906 TI - Pregnancy detection in mice using ultrasound. PMID- 17400907 TI - Biting midges overwintering in Belgium. PMID- 17400908 TI - Older cattle disposal scheme. PMID- 17400909 TI - Withdrawal of routine brucellosis testing in beef herds. PMID- 17400910 TI - Encephalitis in a northern bottlenose whale. PMID- 17400911 TI - Aquatic veterinary medicine. PMID- 17400912 TI - The impact of Epstein-Barr virus status on clinical outcome in diffuse large B cell lymphoma. AB - To define prognostic impact of Epstein-Barr virus (EBV) infection in diffuse large B-cell lymphoma (DLBCL), we investigated EBV status in patients with DLBCL. In all, 380 slides from paraffin-embedded tissue were available for analysis by EBV-encoded RNA-1 (EBER) in situ hybridization, and 34 cases (9.0%) were identified as EBER-positive. EBER positivity was significantly associated with age greater than 60 years (P = .005), more advanced stage (P < .001), more than one extranodal involvement (P = .009), higher International Prognostic Index (IPI) risk group (P = .015), presence of B symptom (P = .004), and poorer outcome to initial treatment (P = .006). The EBER(+) patients with DLBCL demonstrated substantially poorer overall survival (EBER(+) vs EBER(-) 35.8 months [95% confidence interval (CI), 0-114.1 months] vs not reached, P = .026) and progression-free survival (EBER(+) vs EBER(-) 12.8 months [95% CI, 0-31.8 months] vs 35.8 months [95% CI, 0-114.1 months], respectively (P = .018). In nongerminal center B-cell-like subtype, EBER in situ hybridization positivity retained its statistical significance at the multivariate level (P = .045). Nongerminal center B-cell-like patients with DLBCL with EBER positivity showed substantially poorer overall survival with 2.9-fold (95% CI, 1.1-8.1) risk for death. Taken together, DLBCL patients with EBER in situ hybridization+ pursued more rapidly deteriorating clinical course with poorer treatment response, survival, and progression-free survival. PMID- 17400913 TI - Inhibition of Hsp90 attenuates inflammation in endotoxin-induced uveitis. AB - Heat shock protein (Hsp) 90 inhibitors, such as 17-allylamino-17-demethoxy geldanamycin (17-AAG), constitute promising novel therapeutic agents. We investigated the anti-inflammatory activity of 17-AAG in endotoxin-induced uveitis (EIU) in rats. After the induction of EIU with a footpad injection of lipopolysaccharide (LPS), female Lewis rats received a single intraperitoneal. (i.p.) injection of 17-AAG or vehicle. Twenty-four hours later, the retinas were extracted and assayed for leukocyte adhesion; blood-retinal barrier breakdown; VEGF, TNF-alpha, IL-1beta, and CD14 protein levels; NF-kappaB and HIF-1alpha activity; hsp90 and 70 levels and expression and phosphorylation of the tight junction proteins ZO-1 and occludin. 17-AAG treatment significantly suppressed the LPS-induced increase in retinal leukocyte adhesion; vascular leakage; NF kappaB, HIF-1alpha, p38, and PI-3K activity; and VEGF, TNF-alpha, and IL-1beta levels. 17-AAG also suppressed phosphorylation of ZO-1 and occludin by inhibiting their association with p38 and PI-3K. Although 17-AAG treatment did not reduce the LPS-induced increase in total CD14 levels in leukocytes, it significantly decreased membrane CD14 levels. These data suggest that Hsp90 inhibition suppresses several cardinal manifestations of endotoxin-induced uveitis in the rat. 17-AAG has demonstrated a favorable safety profile in clinical trials in cancer patients and represents a promising therapeutic agent for the treatment of inflammatory eye diseases. PMID- 17400914 TI - Gastrointestinal hormones (anorexigenic peptide YY and orexigenic ghrelin) influence neural tube development. AB - Gastrointestinal (GI) hormones play an important role in GI secretion, motility, and eating behaviors. It was recently suggested that GI hormones may have a trophic role in GI tract. Here we demonstrate that two principal GI hormones, anorexigenic peptide YY (PYY) and orexigenic ghrelin, affect neural tube development. Chronic administration into the pregnant mice or transgenic overexpression of PYY led to a neural tube defect (NTD) in the embryos that was blocked by ghrelin. PYY Y1 receptor antagonist prevented the occurrence of NTD induced not only by PYY but also by vitamin A, a well-known teratogen in humans and animals. Y1 receptor deficiency also engendered NTDs, indicating the need to maintain normal Y1 receptor signaling. The present study is the first linking GI hormones to the leading cause of infant mortality and provides a novel insight for neurogenesis in which materno-fetal communication through GI hormones appears to be important. PMID- 17400915 TI - Solution structure of HI1506, a novel two-domain protein from Haemophilus influenzae. AB - HI1506 is a 128-residue hypothetical protein of unknown function from Haemophilus influenzae. It was originally annotated as a shorter 85-residue protein, but a more detailed sequence analysis conducted in our laboratory revealed that the full-length protein has an additional 43 residues on the C terminus, corresponding with a region initially ascribed to HI1507. As part of a larger effort to understand the functions of hypothetical proteins from Gram-negative bacteria, and H. influenzae in particular, we report here the three-dimensional solution NMR structure for the corrected full-length HI1506 protein. The structure consists of two well-defined domains, an alpha/beta 50-residue N-domain and a 3-alpha 32-residue C-domain, separated by an unstructured 30-residue linker. Both domains have positively charged surface patches and weak structural homology with folds that are associated with RNA binding, suggesting a possible functional role in binding distal nucleic acid sites. PMID- 17400916 TI - Crystal structure of the Streptococcus pneumoniae mevalonate kinase in complex with diphosphomevalonate. AB - Streptococcus pneumoniae, a ubiquitous gram-positive pathogen with an alarming, steadily evolving resistance to frontline antimicrobials, poses a severe global health threat both in the community and in the clinic. The recent discovery that diphosphomevalonate (DPM), an essential intermediate in the isoprenoid biosynthetic pathway, potently and allosterically inhibits S. pneumoniae mevalonate kinase (SpMK) without affecting the human isozyme established a new target and lead compound for antimicrobial design. Here we present the crystal structure of the first S. pneumoniae mevalonate kinase, at a resolution of 2.5 A and in complex with DPM.Mg(2+) in the active-site cleft. Structural comparison of SpMK with other members of the GHMP kinase family reveals that DPM functions as a partial bisubstrate analog (mevalonate linked to the pyrophosphoryl moiety of ATP) in that it elicits a ternary-complexlike form of the enzyme, except for localized disordering in a region that would otherwise interact with the missing portion of the nucleotide. Features of the SpMK-binding pockets are discussed in the context of established mechanistic findings and inherited human diseases linked to MK deficiency. PMID- 17400917 TI - Structural evidence for regulation and specificity of flaviviral proteases and evolution of the Flaviviridae fold. AB - Pathogenic members of the flavivirus family, including West Nile Virus (WNV) and Dengue Virus (DV), are growing global threats for which there are no specific treatments. The two-component flaviviral enzyme NS2B-NS3 cleaves the viral polyprotein precursor within the host cell, a process that is required for viral replication. Here, we report the crystal structure of WNV NS2B-NS3pro both in a substrate-free form and in complex with the trypsin inhibitor aprotinin/BPTI. We show that aprotinin binds in a substrate-mimetic fashion in which the productive conformation of the protease is fully formed, providing evidence for an "induced fit" mechanism of catalysis and allowing us to rationalize the distinct substrate specificities of WNV and DV proteases. We also show that the NS2B cofactor of WNV can adopt two very distinct conformations and that this is likely to be a general feature of flaviviral proteases, providing further opportunities for regulation. Finally, by comparing the flaviviral proteases with the more distantly related Hepatitis C virus, we provide insights into the evolution of the Flaviviridae fold. Our work should expedite the design of protease inhibitors to treat a range of flaviviral infections. PMID- 17400918 TI - Solution structure of human sorting nexin 22. AB - The sorting nexins (SNXs) constitute a large group of PX domain-containing proteins that play critical roles in protein trafficking. We report here the solution structure of human sorting nexin 22 (SNX22). Although SNX22 has <30% sequence identity with any PX domain protein of known structure, it was found to contain the alpha/beta fold and compact structural core characteristic of PX domains. Analysis of the backbone dynamics of SNX22 by NMR relaxation measurements revealed that the two walls of the ligand binding cleft undergo internal motions: on the picosecond timescale for the beta1/beta2 loop and on the micro- to millisecond timescale for the loop between the polyproline motif and helix alpha2. Regions of the SNX22 structure that differ from those of other PX domains include the loop connecting strands beta1 and beta2 and the loop connecting helices alpha1 and alpha2, which appear to be more mobile than corresponding loops in other known structures. The interaction of dibutanoyl phosphatidylinositol-3-phosphate (dibutanoyl-PtdIns(3)P) with SNX22 was investigated by an NMR titration experiment, which identified the binding site in a basic cleft and indicated that ligand binding leads only to a local structural rearrangement as has been found with other PX domains. Because motions in the loops are damped out when dibutanoyl-PtdIns(3)P binds, entropic effects could contribute to the lower affinity of SNX22 for this ligand compared to other PX domains. PMID- 17400919 TI - Structural factors affecting the choice between latency transition and polymerization in inhibitory serpins. AB - Plasminogen activator inhibitor-1 (PAI-1), a member of the serine protease inhibitor (serpin) protein family, is unique among the serpins in its conformational lability. This lability allows spontaneous conversion of the active form to a more stable, latent conformation under physiological conditions. In other serpins, polymerization, rather than latency transition, is induced under pathological conditions or upon heat treatment. To identify specific factors promoting latency conversion in PAI-1, we mutated PAI-1 at various positions and compared the effects with those of equivalent mutations in alpha(1) antitrypsin, the archetypal serpin. Mutations that improved interactions with the turn between helix F and the third strand of beta-sheet A (thFs3A) or the fifth strand of beta-sheet A (s5A), which are near the site of latency transition associated insertion of the reactive center loop, retarded latency conversion but did not greatly increase structural stability. Mutations that decreased interactions with s2C facilitated conformational conversion, possibly by releasing the reactive center loop from beta-sheet C. Mutations of Thr93 that filled a hydrophobic surface pocket on s2A dramatically increased structural stability but had a negligible effect on the conformational transition. Our results suggest that the structural features controlling latency transition in PAI-1 are highly localized, whereas the conformational strain of the native forms of other inhibitory serpins is distributed throughout the molecule and induces polymerization. PMID- 17400920 TI - Crystal structure of the MAP kinase binding domain and the catalytic domain of human MKP5. AB - MAP kinase phosphatases (MKPs) have crucial roles in regulating the signaling activity of MAP kinases and are potential targets for drug discovery against human diseases. These enzymes contain a catalytic domain (CD) as well as a binding domain (BD) that help recognize the target MAP kinase. We report here the crystal structures at up to 2.2 A resolution of the BD and CD of human MKP5 and compare them to the known structures from other MKPs. Dramatic structural differences are observed between the BD of MKP5 and that of MKP3 determined previously by NMR. In particular, the cluster of positively charged residues that is important for MAP kinase binding is located in completely different positions in the two structures, with a distance of 25 A between them. Moreover, this cluster is alpha-helical in MKP5, while it forms a loop followed by a beta-strand in MKP3. These large structural differences could be associated with the distinct substrate preferences of these phosphatases, but further studies are needed to confirm this. The CD of MKP5 is observed in an active conformation, and two loops in the active site have backbone shifts of up to 5 A relative to the inactive CDs from other MKPs. PMID- 17400921 TI - Ligand binding and allostery can emerge simultaneously. AB - A heterotropic allosteric effect involves an effector molecule that is distinct from the substrate or ligand of the protein. How heterotropic allostery originates is an unanswered question. We have previously created several heterotropic allosteric enzymes by recombining the genes for TEM1 beta-lactamase (BLA) and maltose binding protein (MBP) to create BLAs that are positively or negatively regulated by maltose. We show here that one of these engineered enzymes has approximately 10(6) M(-1) affinity for Zn(2+), a property that neither of the parental proteins possesses. Furthermore, Zn(2+) is a negative effector that noncompetitively switches off beta-lactam hydrolysis activity. Mutagenesis experiments indicate that the Zn(2+)-binding site does not involve a histidine or a cysteine, which is atypical of natural Zn(2+)-binding sites. These studies also implicate helices 1 and 12 of the BLA domain in allosteric signal propagation. These results support a model for the evolution of heterotropic allostery in which effector affinity and allosteric signaling emerge simultaneously. PMID- 17400922 TI - Crystal structure of the human ephrin-A5 ectodomain. AB - The Eph receptors, the largest subfamily of receptor tyrosine kinases, and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, pathfinding, and mobility in the nervous and cardiovascular systems. Recent structural studies have revealed unique molecular features that explain many of the biochemical and signaling properties of Ephs and ephrins. Nevertheless, open questions remain, including understanding the precise molecular mechanism underlining their binding-partner preferences and subclass specificity. In this study, we have determined and present the crystal structure of the extracellular domain of ephrin-A5-the first structure of an unbound A class ephrin. The structure, determined at 2.1 A resolution, is a variation of the Greek key beta-barrel folding topology, containing eight beta-strands, and stabilized by two disulphide bonds. Overall, ephrin-A5 is structurally very similar to ephrin-B1 and ephrin-B2 but, unlike ephrin-B2, it does not show dimerization either in solution or in the crystals. Comparing free ephrin-A5 to the previously published structure of EphB2-bound ephrin-A5 reveals that significant conformational changes occur only around the G-H ephrin loop that upon binding bends toward the receptor. Interestingly, the G-H loop undergoes a very similar conformational rearrangement in ephrin-B2 upon receptor binding. The results of this study further emphasize the importance of the G-H loop for receptor recognition and selectivity, and could serve as a starting point for the development of structure-based Eph antagonists. PMID- 17400923 TI - Estrogen receptor-ligand complexes measured by chip-based nanoelectrospray mass spectrometry: an approach for the screening of endocrine disruptors. AB - In the present report, a method based on chip-based nanoelectrospray mass spectrometry (nanoESI-MS) is described to detect noncovalent ligand binding to the human estrogen receptor alpha ligand-binding domain (hERalpha LBD). This system represents an important environmental interest, because a wide variety of molecules, known as endocrine disruptors, can bind to the estrogen receptor (ER) and induce adverse health effects in wildlife and humans. Using proper experimental conditions, the nanoESI-MS approach allowed for the detection of specific ligand interactions with hERalpha LBD. The relative gas-phase stability of selected hERalpha LBD-ligand complexes did not mirror the binding affinity in solution, a result that demonstrates the prominent role of hydrophobic contacts for stabilizing ER-ligand complexes in solution. The best approach to evaluate relative solution-binding affinity by nanoESI-MS was to perform competitive binding experiments with 17beta-estradiol (E2) used as a reference ligand. Among the ligands tested, the relative binding affinity for hERalpha LBD measured by nanoESI-MS was 4-hydroxtamoxifen approximately diethylstilbestrol > E2 >> genistein >> bisphenol A, consistent with the order of the binding affinities in solution. The limited reproducibility of the bound to free protein ratio measured by nanoESI-MS for this system only allowed the binding constants (K(d)) to be estimated (low nanomolar range for E2). The specificity of nanoESI-MS combined with its speed (1 min/ligand), low sample consumption (90 pmol protein/ligand), and its sensitivity for ligand (30 ng/mL) demonstrates that this technique is a promising method for screening suspected endocrine disrupting compounds and to qualitatively evaluate their binding affinity. PMID- 17400924 TI - The first step of hen egg white lysozyme fibrillation, irreversible partial unfolding, is a two-state transition. AB - Amyloid fibril depositions are associated with many neurodegenerative diseases as well as amyloidosis. The detailed molecular mechanism of fibrillation is still far from complete understanding. In our previous study of in vitro fibrillation of hen egg white lysozyme, an irreversible partially unfolded intermediate was characterized. A similarity of unfolding kinetics found for the secondary and tertiary structure of lysozyme using deep UV resonance Raman (DUVRR) and tryptophan fluorescence spectroscopy leads to a hypothesis that the unfolding might be a two-state transition. In this study, chemometric analysis, including abstract factor analysis (AFA), target factor analysis (TFA), evolving factor analysis (EFA), multivariate curve resolution-alternating least squares (ALS), and genetic algorithm, was employed to verify that only two principal components contribute to the DUVRR and fluorescence spectra of soluble fraction of lysozyme during the fibrillation process. However, a definite conclusion on the number of conformers cannot be made based solely on the above spectroscopic data although chemometric analysis suggested the existence of two principal components. Therefore, electrospray ionization mass spectrometry (ESI-MS) was also utilized to address the hypothesis. The protein ion charge state distribution (CSD) envelopes of the incubated lysozyme were well fitted with two principal components. Based on the above analysis, the partial unfolding of lysozyme during in vitro fibrillation was characterized quantitatively and proven to be a two state transition. The combination of ESI-MS and Raman and fluorescence spectroscopies with advanced statistical analysis was demonstrated to be a powerful methodology for studying protein structural transformations. PMID- 17400925 TI - Exploring subdomain cooperativity in T4 lysozyme II: uncovering the C-terminal subdomain as a hidden intermediate in the kinetic folding pathway. AB - Intermediates along a protein's folding pathway can play an important role in its biology. Previous kinetics studies have revealed an early folding intermediate for T4 lysozyme, a small, well-characterized protein composed of an N-terminal and a C-terminal subdomain. Pulse-labeling hydrogen exchange studies suggest that residues from both subdomains contribute to the structure of this intermediate. On the other hand, equilibrium native state hydrogen experiments have revealed a high-energy, partially unfolded form of the protein that has an unstructured N terminal subdomain and a structured C-terminal subdomain. To resolve this discrepancy between kinetics and equilibrium data, we performed detailed kinetics analyses of the folding and unfolding pathways of T4 lysozyme, as well as several point mutants and large-scale variants. The data support the argument for the presence of two distinct intermediates, one present on each side of the rate limiting transition state barrier. The effects of circular permutation and site specific mutations in the wild-type and circular permutant background, as well as a fragment containing just the C-terminal subdomain, support a model for the unfolding intermediate with an unfolded N-terminal and a folded C-terminal subdomain. Our results suggest that the partially unfolded form identified by native state hydrogen exchange resides on the folded side of the rate-limiting transition state and is, therefore, under most conditions, a "hidden" intermediate. PMID- 17400926 TI - Exploring subdomain cooperativity in T4 lysozyme I: structural and energetic studies of a circular permutant and protein fragment. AB - Small proteins are generally observed to fold in an apparent two-state manner. Recently, however, more sensitive techniques have demonstrated that even seemingly single-domain proteins are actually made up of smaller subdomains. T4 lysozyme is one such protein. We explored the relative autonomy of its two individual subdomains and their contribution to the overall stability of T4 lysozyme by examining a circular permutation (CP13*) that relocates the N terminal A-helix, creating subdomains that are contiguous in sequence. By determining the high-resolution structure of CP13* and characterizing its energy landscape using native state hydrogen exchange (NSHX), we show that connectivity between the subdomains is an important determinant of the energetic cooperativity but not structural integrity of the protein. The circular permutation results in a protein more easily able to populate a partially unfolded form in which the C terminal subdomain is folded and the N-terminal subdomain is unfolded. We also created a fragment model of this intermediate and demonstrate using X-ray crystallography that its structure is identical to the corresponding residues in the full-length protein with the exception of a small network of hydrophobic interactions. In sum, we conclude that the C-terminal subdomain dominates the energetics of T4 lysozyme folding, and the A-helix serves an important role in coupling the two subdomains. PMID- 17400927 TI - CO migration pathways in cytochrome P450cam studied by molecular dynamics simulations. AB - Previous laser flash photolysis investigations between 100 and 300 K have shown that the kinetics of CO rebinding with cytochrome P450(cam)(camphor) consist of up to four different processes revealing a complex internal dynamics after ligand dissociation. In the present work, molecular dynamics simulations were undertaken on the ternary complex P450(cam)(cam)(CO) to explore the CO migration pathways, monitor the internal cavities of the protein, and localize the CO docking sites. One trajectory of 1 nsec with the protein in a water box and 36 trajectories of 1 nsec in the vacuum were calculated. In each trajectory, the protein contained only one CO ligand on which no constraints were applied. The simulations were performed at 200, 300, and 320 K. The results indicate the presence of seven CO docking sites, mainly hydrophobic, located in the same moiety of the protein. Two of them coincide with xenon binding sites identified by crystallography. The protein matrix exhibits eight persistent internal cavities, four of which corresponding to the ligand docking sites. In addition, it was observed that water molecules entering the protein were mainly attracted into the polar pockets, far away from the CO docking sites. Finally, the identified CO migration pathways provide a consistent interpretation of the experimental rebinding kinetics. PMID- 17400928 TI - Glucose infusion in mice: a new model to induce beta-cell replication. AB - Developing new techniques to induce beta-cells to replicate is a major goal in diabetes research. Endogenous beta-cells replicate in response to metabolic changes, such as obesity and pregnancy, which increase insulin requirement. Mouse genetic models promise to reveal the pathways responsible for compensatory beta cell replication. However, no simple, short-term, physiological replication stimulus exists to test mouse models for compensatory replication. Here, we present a new tool to induce beta-cell replication in living mice. Four-day glucose infusion is well tolerated by mice as measured by hemodynamics, body weight, organ weight, food intake, and corticosterone level. Mild sustained hyperglycemia and hyperinsulinemia induce a robust and significant fivefold increase in beta-cell replication. Glucose-induced beta-cell replication is dose and time dependent. Beta-cell mass, islet number, beta-cell size, and beta-cell death are not altered by glucose infusion over this time frame. Glucose infusion increases both the total protein abundance and nuclear localization of cyclin D2 in islets, which has not been previously reported. Thus, we have developed a new model to study the regulation of compensatory beta-cell replication, and we describe important novel characteristics of mouse beta-cell responses to glucose in the living pancreas. PMID- 17400929 TI - Awakening and counterregulatory response to hypoglycemia during early and late sleep. AB - OBJECTIVE: Nocturnal hypoglycemia represents an important problem for diabetic patients, which has been primarily attributed to an attenuated hormonal counterregulation during sleep. So far, hypoglycemia counterregulation has been exclusively examined during early nocturnal sleep, although early sleep differs markedly in sleep stage architecture from late sleep. Here, we investigated whether awakening and counterregulatory responses differ between early and late sleep. RESEARCH DESIGN AND METHODS: Sixteen healthy subjects were tested on three occasions. On two nights, a linear fall in plasma glucose to a nadir of 2.2 mmol/l within 60 min was induced by insulin infusion. On one night, this was done immediately after sleep onset and on the other night after approximately 3.5 h of sleep. In a further control night, no hypoglycemia was induced. RESULTS: During early sleep, 10 subjects awoke in response to hypoglycemia, whereas no subject awoke during the corresponding interval of the control night (P < 0.004). During late sleep, all subjects awoke upon hypoglycemia, and four subjects awoke spontaneously during the corresponding control interval (P < 0.001). The pattern indicates that the frequency of awakenings caused by hypoglycemia is similar for early and late sleep. Increases in epinephrine, norepinephrine, ACTH, cortisol, and growth hormone were distinctly weaker during late than early hypoglycemia (all P < 0.05). CONCLUSIONS: Diminished hormonal counterregulation during late sleep could be one factor contributing to the clinically observed accumulation of hypoglycemic episodes in the later part of the night in patients with diabetes. PMID- 17400930 TI - Reactive oxygen species as a signal in glucose-stimulated insulin secretion. AB - One of the unique features of beta-cells is their relatively low expression of many antioxidant enzymes. This could render beta-cells susceptible to oxidative damage but may also provide a system that is sensitive to reactive oxygen species as signals. In isolated mouse islets and INS-1(832/13) cells, glucose increases intracellular accumulation of H2O2. In both models, insulin secretion could be stimulated by provision of either exogenous H2O2 or diethyl maleate, which raises intracellular H2O2 levels. Provision of exogenous H2O2 scavengers, including cell permeable catalase and N-acetyl-L-cysteine, inhibited glucose-stimulated H2O2 accumulation and insulin secretion (GSIS). In contrast, cell permeable superoxide dismutase, which metabolizes superoxide into H2O2, had no effect on GSIS. Because oxidative stress is an important risk factor for beta-cell dysfunction in diabetes, the relationship between glucose-induced H2O2 generation and GSIS was investigated under various oxidative stress conditions. Acute exposure of isolated mouse islets or INS-1(832/13) cells to oxidative stressors, including arsenite, 4-hydroxynonenal, and methylglyoxal, led to decreased GSIS. This impaired GSIS was associated with increases in a battery of endogenous antioxidant enzymes. Taken together, these findings suggest that H2O2 derived from glucose metabolism is one of the metabolic signals for insulin secretion, whereas oxidative stress may disturb its signaling function. PMID- 17400931 TI - Implantation site-dependent dysfunction of transplanted pancreatic islets. AB - OBJECTIVE: Clinical islet transplantations are performed through infusion of islets via the portal vein into the liver. This study aimed at characterizing the influence of the implantation microenvironment on islet graft metabolism and function. RESEARCH DESIGN AND METHODS: Islets were transplanted into their normal environment, i.e., the pancreas, or intraportally into the liver of mice. One month posttransplantation, the transplanted islets were retrieved and investigated for changes in function and gene expression. RESULTS: Insulin content, glucose-stimulated insulin release, (pro)insulin biosynthesis, and glucose oxidation rate were markedly decreased in islets retrieved from the liver, both when compared with islets transplanted into the pancreas and endogenous islets. Islets transplanted into the pancreas showed normal insulin content, (pro)insulin biosynthesis, and glucose oxidation rate but increased basal insulin secretion and impaired glucose stimulation index. Gene expression data for retrieved islets showed downregulation of pancreatic and duodenal homeobox gene-1, GLUT-2, glucokinase, mitochondrial glycerol-phosphate dehydrogenase, and pyruvate carboxylase, preferentially in intraportally transplanted islets. CONCLUSIONS: Islets transplanted into their normal microenvironment, i.e., the pancreas, display gene expression changes when compared with endogenous islets but only moderate changes in metabolic functions. In contrast, site-specific properties of the liver markedly impaired the metabolic functions of intraportally transplanted islets. PMID- 17400932 TI - Sociodemographic and psychosocial correlates of smoking-induced deprivation and its effect on quitting: findings from the International Tobacco Control Policy Evaluation Survey. AB - AIMS: To determine the prevalence and characteristics of smokers who experience smoking-induced deprivation (SID), and to examine its effect on quit attempts, relapse and cessation. METHODS: Waves 2 and 3 (2003-5) of the International Tobacco Control Policy Evaluation Survey were used, which is a prospective study of a cohort of smokers in the US, Canada, UK and Australia. SID was measured with the question "In the last six months, have you spent money on cigarettes that you knew would be better spent on household essentials like food?" A total of 7802 smokers participated in the survey in wave 2, of whom 5408 were also interviewed in wave 3. FINDINGS: The proportion of smokers who reported SID was highest in Australia (33%) and lowest in the UK (20%). Younger age, minority status and low income were associated with a higher probability of SID. Some of the other factors related to a higher probability of SID were higher level of nicotine dependence, having an intention to quit, and smoking to help one socialise or control weight. The relationship between SID and quit attempt was mediated by having an intention to quit and worrying that smoking would damage health and reduce the quality of life. The relationship between SID and relapse was mediated by perceived stress. SID was not associated with successful cessation. CONCLUSIONS: Many smokers experience deprivation that is the result of their smoking. Strategies to reduce the prevalence of smoking probably effect a general improvement in standards of living and reduction in deprivation. PMID- 17400933 TI - Asian herbal-tobacco cigarettes: "not medicine but less harmful"? AB - OBJECTIVE: To describe the development and health claims of Asian herbal-tobacco cigarettes. METHODS: Analysis of international news sources, company websites, and the transnational tobacco companies' (TTC) documents. PubMed searches of herbs and brands. RESULTS: Twenty-three brands were identified, mainly from China. Many products claimed to relieve respiratory symptoms and reduce toxins, with four herb-only products advertised for smoking cessation. No literature was found to verify the health claims, except one Korean trial of an herb-only product. Asian herbal-tobacco cigarettes were initially produced by China by the 1970s and introduced to Japan in the 1980s. Despite initial news about research demonstrating a safer cigarette, the TTC analyses of these cigarettes suggest that these early products were not palatable and had potentially toxic cardiovascular effects. By the late 1990s, China began producing more herbal tobacco cigarettes in a renewed effort to reduce harmful constituents in cigarettes. After 2000, tobacco companies from Korea, Taiwan, and Thailand began producing similar products. Tobacco control groups in Japan, Taiwan, and Thailand voiced concern over the health claims of herbal-tobacco products. In 2005, China designated two herbal-tobacco brands as key for development. CONCLUSION: Asian herbal-tobacco cigarettes claim to reduce harm, but no published literature is available to verify these claims or investigate unidentified toxicities. The increase in Asian herbal-tobacco cigarette production by 2000 coincides with the Asian tobacco companies' regular scientific meetings with TTCs and their interest in harm reduction. Asia faces additional challenges in tobacco control with these culturally concordant products that may discourage smokers from quitting. PMID- 17400934 TI - Evidence, ethics, hubris and the future of second-hand smoke policy. PMID- 17400936 TI - Canada: pharmacy tobacco sales to mentally ill people. PMID- 17400943 TI - Women's smoking history prior to entering the US Navy: a prospective predictor of performance. AB - OBJECTIVE: To examine whether women's tobacco use prior to entering the US Navy is predictive of subsequent career performance. A priori predictions were that smoking at entry into the Navy would be related to early attrition, poorer job performance, more disciplinary problems and lower likelihood of re-enlistment. METHODS: A prospective cohort analysis of 5487 women entering the US Navy between March 1996 and March 1997 was conducted. Navy attrition/retention and career performance measures, such as time in service, early attrition, type of discharge, misconduct, number of promotions, demotions and unauthorised absences, highest paygrade achieved, and re-enlistment were examined. RESULTS: Compared with never smokers, daily smokers at entry into the US Navy had subsequent career outcomes consistently indicating poorer job performance (eg, early attrition prior to serving a full-term enlistment, more likely to have a less-than honourable discharge, more demotions and desertions, lower achieved paygrade and less likely to re-enlist). Other types of smokers consistently fell between never and daily smokers on career outcome measures. CONCLUSIONS: For women entering the US Navy, being a daily smoker is a prospective predictor of poorer performance in the Navy. Future research should evaluate the effectiveness of cessation intervention with smoker-enlistees prior to their entering the Navy, to assess the impact on subsequent career outcomes. PMID- 17400944 TI - The California Tobacco Control Program's effect on adult smokers: (1) Smoking cessation. AB - OBJECTIVES: To estimate national population trends in long-term smoking cessation by age group and to compare cessation rates in California (CA) with those of two comparison groups of states. SETTING: Retrospective smoking history of a population sample from the US: from CA, with a comprehensive tobacco-control programme since 1989 with the goal of denormalising tobacco use; from New York and New Jersey (NY & NJ), with similar high cigarette prices but no comprehensive programme; and from the tobacco-growing states (TGS), with low cigarette prices, no tobacco-control programme and social norms relatively supportive of tobacco use. PARTICIPANTS: Respondents to the Current Population Survey-Tobacco Use Supplements (1992-2002; n = 57 918 non-Hispanic white ever-smokers). MAIN OUTCOME MEASURES: The proportion of recent ever-smokers attaining long-term abstinence (quit > or = 1 year) and the successful-quit ratio (the proportion of all ever smokers abstinent > or = 1 year). RESULTS: Nationally, long-term cessation rates increased by 25% from the 1980s to the 1990s, averaging 3.4% per year in the 1990s. Cessation increased for all age groups, and by > 40% (p<0.001) among smokers aged 20-34 years. For smokers aged < 50 years, higher cigarette prices were associated with higher quitting rates. For smokers aged < 35 years, quitting rates in CA were higher than in either comparison group (p<0.05). Half of the ever-smokers had quit smoking by age 44 years in CA, 47 years in NY & NJ, and by age 54 years in TGS. CONCLUSION: Successful smoking cessation increased by 25% during the 1990s in the US. Comprehensive tobacco-control programmes were associated with greater cessation success than were with high cigarette prices alone, although both effects were limited to younger adults. PMID- 17400945 TI - The California Tobacco Control Program's effect on adult smokers: (2) Daily cigarette consumption levels. AB - OBJECTIVE: To investigate the association of the California Comprehensive Tobacco Control Program with self-reported population trends of cigarette consumption during 1992-2002. SETTING AND PARTICIPANTS: Participants were non-Hispanic white daily smokers (aged 20-64 years, n = 24 317) from the Tobacco Use Supplements to the Current Population Survey (1992-2002). We compared age-specific trends in consumption among daily smokers in three groups of states with differing tobacco control initiatives: California (CA; high cigarette price/comprehensive programme), New York and New Jersey (high cigarette price/no comprehensive programme), and tobacco-growing states (TGS; low cigarette price/no comprehensive programme). RESULTS: There was a general decline in cigarette consumption across all age groups in each category of states between 1992 and 2002, except the oldest age group in the TGS. The largest annual decline in the average number of cigarettes per day was observed among daily smokers in CA who were aged > or = 35 years (-0.41 cigarettes/day/year (95% CI -0.52 to -0.3)). This rate was significantly higher than the -0.22 cigarettes/day/year (95% CI -0.3 to -0.16; p<0.02) observed in same-age daily smokers from New York and New Jersey, and significantly higher than the rate in same-age daily smokers from the TGS (-0.15 cigarettes/day/year (95% CI -0.22 to -0.08; p<0.002)). There were no significant differences across state groups in the decline observed in daily smokers aged 20 34 years. In 2002, only 12% of daily smokers in CA smoked more than a pack per day, which was significantly lower than the 17% in New York and New Jersey, which again was significantly lower than the 25% in the TGS. CONCLUSIONS: The California Tobacco Control Program was associated with significant declines in cigarette consumption among daily smokers aged > or = 35 years of age, which in turn should lead to declines in tobacco-related health effects. The decline in consumption among young adult smokers was a national trend. PMID- 17400946 TI - The California Tobacco Control Program's effect on adult smokers: (3) Similar effects for African Americans across states. AB - OBJECTIVE: To compare trends in African-American (AA) and non-Hispanic white (NHW) smoking between states categorised as having three different levels of tobacco control practice in the 1990s. SETTING AND PARTICIPANTS: Analysis of 1992 3 to 2001-2 Tobacco Use Supplements to the Current Population Survey for differences in adult (20-64 years) daily smoking prevalence for AAs and NHWs across states: California (CA; high cigarette price/comprehensive programme), New York (NY) and New Jersey (NJ; high cigarette price/no comprehensive programme), and tobacco growing states (TGS; low cigarette price/no comprehensive programme). RESULTS: From 1992-3 to 2001-2, there were large declines in AA smoking across states (2.7-3.8% decrease/year, adjusted for age, income, education, gender; p<0.05). Adjusted NHW smoking prevalence declined significantly only in CA. AA prevalence declined significantly and did not differ across state groups. In all years, in all state groups, adjusted prevalence was either not significantly different or was lower for AAs than for NHWs. More recent cohorts of AAs appeared to have taken up smoking at lower rates than older cohorts. CONCLUSION: There were uniformly large declines in AA smoking from 1992-3 to 2001-2 across states, independent of type of tobacco control strategy. Further research is needed into factors associated with smoking declines among AAs. PMID- 17400947 TI - Effect of smoking regulations in local restaurants on smokers' anti-smoking attitudes and quitting behaviours. AB - OBJECTIVE: To examine the effect of smoking regulations in local restaurants on anti-smoking attitudes and quitting behaviours among adult smokers. DESIGN: Hierarchical linear modelling (HLM) was used to assess the relationship between baseline strength of town-level restaurant smoking regulation and follow-up (1) perceptions of the social acceptability of smoking and (2) quitting behaviours. SETTING: Each of the 351 Massachusetts towns was classified as having strong (complete smoking ban) or weak (all other and no smoking restrictions) restaurant smoking regulations. SUBJECTS: 1712 adult smokers of Massachusetts aged > or = 18 years at baseline who were interviewed via random-digit-dial telephone survey in 2001-2 and followed up 2 years later. MAIN OUTCOME MEASURES: Perceived social acceptability of smoking in restaurants and bars, and making a quit attempt and quitting smoking. RESULTS: Among adult smokers who had made a quit attempt at baseline, living in a town with a strong regulation was associated with a threefold increase in the odds of making a quit attempt at follow-up (OR = 3.12; 95% CI 1.51 to 6.44). Regulation was found to have no effect on cessation at follow-up. A notable, although marginal, effect of regulation was observed for perceiving smoking in bars as socially unacceptable only among smokers who reported at baseline that smoking in bars was socially unacceptable. CONCLUSIONS: Although local restaurant smoking regulations did not increase smoking cessation rates, they did increase the likelihood of making a quit attempt among smokers who had previously tried to quit, and seem to reinforce anti-social smoking norms among smokers who already viewed smoking in bars as socially unacceptable. PMID- 17400948 TI - Mortality and life expectancy in relation to long-term cigarette, cigar and pipe smoking: the Zutphen Study. AB - STUDY OBJECTIVE: To study the effect of long-term smoking on all-cause and cause specific mortality, and to estimate the effects of cigarette and cigar or pipe smoking on life expectancy. DESIGN: A long-term prospective cohort study. SETTING: Zutphen, The Netherlands. PARTICIPANTS: 1373 men from the Zutphen Study, born between 1900 and 1920 and studied between 1960 and 2000. MEASUREMENTS: Hazard ratios for the type of smoking, amount and duration of cigarette smoking, obtained from a time-dependent Cox regression model. Absolute health effects of smoking are expressed as differences in life expectancy and the number of disease free years of life. MAIN RESULTS: Duration of cigarette smoking was strongly associated with mortality from cardiovascular disease, lung cancer and chronic obstructive pulmonary disease, whereas both the number of cigarettes smoked as well as duration of cigarette smoking were strongly associated with all-cause mortality. Average cigarette smoking reduced the total life expectancy by 6.8 years, whereas heavy cigarette smoking reduced the total life expectancy by 8.8 years. The number of total life-years lost due to cigar or pipe smoking was 4.7 years. Moreover, cigarette smoking reduced the number of disease-free life-years by 5.8 years, and cigar or pipe smoking by 5.2 years. Stopping cigarette smoking at age 40 increased the life expectancy by 4.6 years, while the number of disease free life-years was increased by 3.0 years. CONCLUSIONS: Cigar or pipe smoking reduces life expectancy to a lesser extent than cigarette smoking. Both the number of cigarettes smoked and duration of smoking are strongly associated with mortality risk and the number of life-years lost. Stopping smoking after age 40 has major health benefits. PMID- 17400949 TI - Does smoking increase sick leave? Evidence using register data on Swedish workers. AB - OBJECTIVE: To examine the effect of smoking on sick leave. METHODS: Nationally representative data on 14,272 workers aged 16-65 years from the 1988-91 waves of the Swedish Survey of Living Conditions were used for the analyses. The data are linked to register-based data, on the annual number of absences due to sickness, from the National Board of Social Insurance. As outcome variable, the annual number of days of sick leave was used. This outcome was analysed as a function of smoking status and an extensive number of control variables, including occupational risk factors, work characteristics and health status. RESULTS: Smoking was found to increase the annual number of days of absence by 10.7 compared with never smoking. Controlling for risk factors at work, and thereby accounting for some of the selection of smokers into riskier jobs, reduced the effect to 9.7 days, corresponding to 38% of all annual absences due to sickness. Moreover, controlling for health status further reduced the effect of smoking to 7.7 days. The effect of smoking on sick leave was similar for men and women. CONCLUSIONS: Smoking showed a large positive effect on the annual number of sick leaves. Hence, the results suggest that the gains to preventing and/or reducing smoking, in terms of reduced production losses, may be large. However, since the large effect of smoking persists when controlling for a range of health factors and occupational factors, the results also suggest that much of the higher number of absences among smokers may be explained by factors other than reduced health. The estimates should be viewed as upper bounds of the effect of smoking on sick leave, since smoking is potentially an endogenous variable. PMID- 17400950 TI - Effect of local youth-access regulations on progression to established smoking among youths in Massachusetts. AB - OBJECTIVE: To test whether community-level restrictions on youth access to tobacco (including both ordinances and enforcement) are associated with less smoking initiation or less progression to established smoking among adolescents. DESIGN: Prospective cohort study of a random sample of adolescents in Massachusetts whose smoking status was assessed by telephone interviews at baseline and 2-year follow-up, and linked to a state-wide database of town-level youth-access ordinances and enforcement practices. PARTICIPANTS: A random sample of 2623 adolescents aged 12-17 years who lived in 295 towns in Massachusetts in 2001-2 and were followed in 2003-4. MAIN OUTCOME MEASURES: The relationship between the strength of local youth access restrictions (including both ordinances and level of enforcement) and (1) never-smokers' smoking initiation rates and (2) experimenters' rate of progression to established smoking was tested in a multilevel analysis that accounted for town-level clustering and adjusted for potential individual, household and town-level confounders. RESULTS: Over 2 years, 21% of 1986 never-smokers initiated smoking and 25% of 518 experimenters became established smokers. The adjusted odds ratio (OR) for smoking initiation was 0.89 (95% CI 0.61 to 1.31) for strong versus weak youth access policies and 0.93 (95% CI 0.67 to 1.29) for medium versus weak policies. The adjusted OR for progression to established smoking among adolescents who had experimented with smoking was 0.79 (95% CI 0.45 to 1.39) for strong versus weak local smoking restrictions and 0.85 (95% CI 0.50 to 1.45) for medium versus weak restrictions. CONCLUSIONS: This prospective cohort study found no association between community-level youth-access restrictions and adolescents' rate of smoking initiation or progression to established smoking over 2 years. PMID- 17400951 TI - Secondhand smoke levels in Scottish pubs: the effect of smoke-free legislation. AB - OBJECTIVE: To compare levels of particulate matter, as a marker of secondhand smoke (SHS) levels, in pubs before and 2 months after the implementation of Scottish legislation to prohibit smoking in substantially enclosed public places. DESIGN: Comparison of SHS levels before and after the legislation in a random selection of 41 pubs in 2 Scottish cities. METHODS: Fine particulate matter <2.5 microm in diameter (PM2.5) was measured discreetly for 30 min in each bar on 1 or 2 visits in the 8 weeks preceding the starting date of the Smoking, Health and Social Care (Scotland) Act 2005 and then again 2 months after the ban. Repeat visits were undertaken on the same day of the week and at approximately the same time of the day. RESULTS: PM2.5 levels before the introduction of the legislation averaged 246 microg/m3 (range 8-902 microg/m3). The average level reduced to 20 microg/m3 (range 6-104 microg/m3) in the period after the ban. Levels of SHS were reduced in all 53 post-ban visits, with the average reduction being 86% (range 12 99%). PM2.5 concentrations in most pubs post-ban were comparable to the outside ambient air PM2.5 level. CONCLUSIONS: This study has produced the largest dataset of pre- and post-ban SHS levels in pubs of all worldwide smoke-free legislations introduced to date. Our results show that compliance with the Smoking, Health and Social Care (Scotland) Act 2005 has been high and this has led to a marked reduction in SHS concentrations in Scottish pubs, thereby reducing both the occupational exposure of workers in the hospitality sector and that of non smoking patrons. PMID- 17400952 TI - British American Tobacco's tactics during China's accession to the World Trade Organization. AB - BACKGROUND: China entered the World Trade Organization (WTO) in 2001 after years of negotiations. As a WTO member, China had to reduce tariffs on imported cigarettes and remove non-tariff barriers to allow foreign cigarettes to be more competitive in the Chinese market. Among foreign tobacco companies, British American Tobacco (BAT) was the most active lobbyist during China's WTO negotiations. OBJECTIVE: To review and analyse BAT's tactics and activities relating to China's entry into the WTO. METHODS: Internal tobacco industry documents were reviewed and are featured here. Industry documents were searched mainly on the website of BAT's Guildford Depository and other documents' websites. 528 documents were evaluated and 142 were determined to be relevant to China's entry into the WTO. RESULTS: BAT was extremely active during the progress of China's entry into the WTO. The company focused its lobbying efforts on two main players in the negotiations: the European Union (EU) and the US. Because of the negative moral and health issues related to tobacco, BAT did not seek public support from officials associated with the WTO negotiations. Instead, BAT lobbyists suggested that officials protect the interests of BAT by presenting the company's needs as similar to those of all European companies. During the negotiation process, BAT officials repeatedly spoke favourably of China's accession into the WTO, with the aim of presenting BAT as a facilitator in this process and of gaining preferential treatment from their Chinese competitor. CONCLUSIONS: BAT's activities clearly suggest that tobacco companies place their own interests above public health interests. Today, China struggles with issues of tobacco control that are aggravated by the aggressive practices of transnational tobacco companies, tobacco-tariff reductions and the huge number of smokers. For the tobacco-control movement to progress in China, health advocates must understand how foreign tobacco companies have undermined anti-tobacco activities by taking advantage of trade liberalisation policies. China should attach importance to public health and comprehensive tobacco-control policies and guarantee strong protection measures from national and international tobacco interests supported by international trade agreements. PMID- 17400953 TI - Nicotine pharmacokinetics and subjective effects of three potential reduced exposure products, moist snuff and nicotine lozenge. AB - OBJECTIVE: To compare nicotine pharmacokinetics and subjective effects of three new smokeless tobacco potential reduced exposure products (PREPs; Ariva, Revel and Stonewall) with moist snuff (Copenhagen) and medicinal nicotine (Commit lozenge). METHODS: 10 subjects completed a randomised, within-subject, crossover study. Subjects used one product for 30 min at each of the five laboratory sessions. Maximal nicotine concentration (Cmax) was determined and area under the concentration time curve (AUC) was calculated for a 90-min period (during use and 60 min after use). Nicotine craving, withdrawal symptoms and ratings of product effects and liking were measured during product use. RESULTS: Nicotine AUC and Cmax were higher for Copenhagen than for any other product (p<0.002) and higher for Commit than for either Ariva or Revel (p<0.001). Cmax for Commit was also higher than for Stonewall (p = 0.03). Craving was lowest during use of Copenhagen (p<0.03). Craving during use of Stonewall, Ariva and Commit was lower than during use of Revel (p<0.05). Withdrawal symptom score during use of Copenhagen was lower than during use of Revel (p = 0.009). Copenhagen scores were higher (p<0.005) than all other products in several measures of drug effects and liking (feel good effects, satisfaction, liking and desire for product, and strength of product). CONCLUSION: The new smokeless tobacco PREPs result in lower nicotine concentrations and equivalent or lower reductions in subjective measures compared with medicinal nicotine. Since health effects of PREPs are largely unknown, medicinal nicotine should be preferentially encouraged for smokers or smokeless tobacco users wishing to switch to lower-risk products. PMID- 17400954 TI - Are homeless transitional shelters receptive to environmental tobacco control interventions? PMID- 17400956 TI - On "The influence of otolith dysfunction..." Murray et al. Phys ther. 2007;87:143 152. PMID- 17400965 TI - Tissue weights and body composition of two genetic lines of barrows and gilts from twenty to one hundred twenty-five kilograms of body weight. AB - Barrows and gilts of 2 genetic lines with differing lean gain potentials (high lean = 375 g of fat-free lean/d; low-lean = 280 g of fat-free lean/d) were used to determine tissue and organ weights and compositions from 20 to 125 kg of BW. The experiment was a 2 (genetic line) x 2 (sex) x 5 (BW) factorial arrangement of treatments in a completely randomized design conducted with 2 groups of pigs in 6 replicates (n = 120 pigs). Six pigs from each sex and genetic line were slaughtered at 20 kg of BW and at 25 kg of BW intervals to 125 kg of BW. At slaughter, the internal tissues and organs were weighed. Loin and ham muscles were dissected from the carcass and trimmed of skin and external fat, and the ham was deboned. Residuals from the loin and ham were combined with the remaining carcass. Body components were ground, and their compositions were determined. The results demonstrated that tissue weights increased (P < 0.01) as BW increased. Loin and ham muscle weights increased but at a greater rate in the high-lean line and in gilts resulting in genetic line x BW and sex x BW interactions (P < 0.01). Liver and heart expressed on a BW or a percentage of empty BW basis increased at a greater rate in the high-lean line resulting in a genetic line x BW interaction (P < 0.01). Liver and intestinal tract weights were heavier in barrows than in gilts, significant only at 45 (P < 0.05), 75 (P < 0.01), and 100 (P < 0.05) kg of BW. Loin and ham muscles from the high-lean genetic line and gilts had greater (P < 0.01) water, protein, and ash contents compared with the low-lean genetic line and barrows resulting in genetic line x BW and sex x BW interactions (P < 0.01). The remaining carcass (minus loin and ham muscles) had greater (P < 0.01) amounts of water and protein, and less (P < 0.01) fat in the high-lean genetic line and gilts. The high-lean genetic line and gilts had more total body water, protein, and ash, but less body fat, with these differences diverging as BW increased, resulting in a genetic line x BW interaction (P < 0.01). The results indicated that liver and heart weights were greater in high-lean pigs, reflecting the greater amino acid metabolism, whereas the liver and intestinal tract weights were greater in barrow than gilts, reflecting their greater feed intakes and metabolism of total nutrients consumed. PMID- 17400966 TI - Comparison of progestin-based estrus synchronization protocols before fixed-time artificial insemination on pregnancy rate in beef heifers. AB - The objective of the experiment was to compare pregnancy rates resulting from fixed-time AI after administration of either 1 of 2 controlled internal drug release (CIDR)-based protocols. Heifers at 3 locations (location 1, n = 78; location 2, n = 61; and location 3, n = 78) were assigned to 1 of 2 treatments within reproductive tract scores (1 = immature to 5 = cycling) by age and BW. Heifers assigned to CIDR Select received a CIDR insert (1.38 g of progesterone) from d 0 to 14 followed by GnRH (100 mug, i.m.) 9 d after CIDR removal (d 23) and PGF2alpha (PG, 25 mg, i.m.) 7 d after GnRH treatment (d 30). Heifers assigned to CO-Synch + CIDR were administered GnRH and received a CIDR insert on d 23 and PG and CIDR removal on d 30. Heifers at location 1 were fitted with a HeatWatch estrus detection system transmitter from the time of PG until 24 d after fixed time AI to allow for continuous estrus detection. Artificial insemination was performed at predetermined fixed times for heifers in both treatments at 72 or 54 h after PG for the CIDR Select and CO-Synch + CIDR groups, respectively. All heifers were administered GnRH at the time of AI. Blood samples were collected 10 d before and immediately before treatment initiation (d 0) to determine pretreatment estrous cyclicity (progesterone > or = 0.5 ng/mL). At location 1, the estrous response during the synchronized period was greater (P = 0.06; 87 vs. 69%, respectively), and the variance for interval to estrus after PG was reduced among CIDR Select- (P < 0.01) compared with CO-Synch + CIDR-treated heifers. Fixed-time AI pregnancy rates were significantly greater (P = 0.02) after the CIDR Select protocol (62%) compared with the CO-Synch + CIDR protocol (47%). In summary, the CIDR Select protocol resulted in a greater and more synchronous estrous response and significantly greater fixed-time AI pregnancy rates compared with the CO-Synch + CIDR protocol. PMID- 17400967 TI - Nutritional value for swine of extruded corn and corn fractions obtained after dry milling. AB - The experiment was designed to assess whether corn fractions or extrusion of corn can result in feed ingredients with a greater nutritional value than corn. Corn grain (8.0% CP, 0.21% P, 9.8% NDF) was processed by extrusion (82.8 degrees C, 345 kPa steam pressure for 12 s) or by dry milling to derive fractions rich in germ (13.1% CP, 1.19% P, 17.2% NDF), hulls (8.1% CP, 0.27% P, 32.6% NDF), and endosperm, namely tails (6.6% CP, 0.07% P, 3.6% NDF) and throughs (7.4% CP, 0.15% P, 4.5% NDF). Relative recovery in each fraction was 16, 20, 44, and 20%, respectively. Ileal digestibility of DM, P, and amino acids was determined using diets containing 7.0% CP from soybean meal and 5.3% CP from one of the test products. To allow for determination of standardized ingredient, ileal digestibility, basal endogenous AA losses were determined using a protein-free diet (74.6% cornstarch and 18.7% sucrose). Soybean meal ileal digestibility was determined using a diet (12.3% CP) based on soybean meal (23.3%). Eight barrows (27 +/- 2 kg) fitted with T-cannulas were fed 8 experimental diets (5-d adaptation and 2-d collection period) such that each diet was evaluated in at least 5 barrows. Relative to corn (77.9 +/- 1.2%), ileal digestibility of DM was greater for extruded corn (82.5%; P = 0.02), tails (85.9%; P < 0.01), and throughs (85.0%; P < 0.01), but it was lower for hulls (62.2%; P < 0.01) and germ (51.1%; P < 0.01). For P, corn (41.6 +/- 9.5%), throughs (47.2%), and hulls (57.3%) had similar ileal digestibility, but germ (7.9%) had lower ileal digestibility (P = 0.02) than corn; tails (27.6%) and extruded corn (23.5%) were not different from corn or germ but were lower than throughs and hulls. For total AA, corn (84.7 +/- 2.4%), throughs (84.3%), and hulls (85.8%) had similar ileal digestibility, but germ (76.6%) had lower ileal digestibility (P < 0.01) than corn; tails (82.0%) and extruded corn (81.7%) were intermediate. In conclusion, germ and hulls have a low ileal DM digestibility; germ also has low AA and P digestibility. Extrusion improved the ileal DM digestibility of corn. To maximize the ileal digestibility, removal of germ and hull from corn or extrusion of corn may thus be of interest. PMID- 17400968 TI - Starch digestion by feedlot cattle: predictions from analysis of feed and fecal starch and nitrogen. AB - To evaluate the utility of N as a digestion marker to predict total tract starch digestion, data from 32 metabolism trials involving 147 steers and 637 individual starch digestibility measurements were compiled. All trials were conducted at the University of California Desert Research and Extension Center. Total tract starch digestibility was determined from concentrations of starch and chromic oxide in feed and feces. In all trials, the steers were adapted to diets for 10 d followed by 4 d for collection of samples of feces. During collection, fecal samples (approximately 200 g, wet basis) were obtained twice daily. Samples from each steer within each collection period were composited for analysis. Diets contained 46.5 +/- 7.4% starch and 1.85 +/- 0.20% N. Apparently digestible N as a percentage of diet DM was closely associated (r(2) = 0.73; P < 0.001) with dietary N concentration. Fecal N concentration (FN, % of DM) explained 35% of the variation in fecal DM excretion (S(y.x) = 4.3; P < 0.001). Incorporating FN into the model, starch digestion was estimated as follows: starch digestion, % of intake = 100 {1 - [(0.938 -0.497FN + 0.0853FN(2)) FS/DS]}, where FS is fecal starch concentration (% of DM) and DS is dietary starch concentration (% of DM; r(2) = 0.94; S(y.x) = 0.68; P < 0.001). Fecal starch concentration alone explained 96% of the variation (S(y.x) = 0.45; P < 0.001) in total tract starch digestion: starch digestion, % = 99.9 - 0.413FS -0.0104FS(2). Omitting cases in our data set in which observed total tract starch digestion was less than 95%, the r(2) between FS and starch digestibility decreased to 0.82 (S(y.x) = 0.26; n = 529). However, estimated starch digestion using the equation incorporating FN remained closely associated with the observed starch digestion (r(2) = 0.90; S(y.x) = 0.22; P < 0.001; n = 529). Equations also were developed to predict NE(m) and NE(g) concentrations of common feed grains based on starch digestibility and FS. Starch digestion can be accurately predicted based on FS. However, incorporation of FN into the model markedly enhanced the estimates of grain quality and the efficacy of processing when total tract starch digestion exceeded 95%. PMID- 17400969 TI - Comparison of progestin-based protocols to synchronize estrus and ovulation before fixed-time artificial insemination in postpartum beef cows. AB - This experiment was designed to compare pregnancy rates in postpartum beef cows resulting from fixed-time AI (FTAI) after treatment with 1 of 2 protocols to synchronize estrus and ovulation. Cross-bred, suckled beef cows (n = 650) at 4 locations (n = 210; n = 158; n = 88; and n = 194) were assigned within a location to 1 of 2 protocols within age group by days postpartum and BCS. Cows assigned to the melengestrol acetate (MGA) Select treatment (MGA Select; n = 327) were fed MGA (0.5 mg x head(-1) x d(-1)) for 14 d, GnRH (100 microg of Cystorelin i.m.) was injected on d 26, and prostaglandin F2alpha (PG; 25 mg of Lutalyse i.m.) was injected on d 33. Cows assigned to the CO-Synch + controlled internal drug release (CIDR) protocol (CO-Synch + CIDR; n = 323) were fed a carrier for 14 d, were injected with GnRH and equipped with an EAZI-BREED CIDR insert (1.38 g of progesterone, Pfizer Animal Health, New York, NY) 12 d after carrier removal, and PG (25 mg of Lutalyse i.m.) was injected and the CIDR were removed on d 33. Fixed time AI was performed at 72 or 66 h after PG for the MGA Select or CO-Synch + CIDR groups, respectively. All cows were injected with GnRH (100 microg of Cystorelin i.m.) at the time of insemination. Blood samples were collected 8 and 1 d before the beginning of MGA or carrier to determine estrous cyclicity status of the cows (estrous cycling vs. anestrus) before treatment [progesterone > or = 0.5 ng/mL (MGA Select, 185/327, 57%; CO-Synch + CIDR, 177/323, 55%); P = 0.65]. There was no difference (P = 0.20) in pregnancy rate to FTAI between treatments (MGA Select, 201/327, 61%; CO-Synch + CIDR, 214/323, 66%). There was also no difference (P = 0.25) between treatments in final pregnancy rate at the end of the breeding period (MGA Select, 305/327, 93%; CO-Synch + CIDR, 308/323, 95%). These data indicate that pregnancy rates to FTAI were comparable after administration of the MGA Select or CO-Synch + CIDR protocols. Both protocols provide opportunities for beef producers to utilize AI and potentially eliminate the need to detect estrus. PMID- 17400970 TI - Phenotypic measurements and various indices of lean and fat tissue development in barrows and gilts of two genetic lines from twenty to one hundred twenty-five kilograms of body weight. AB - Two genetic lines with different lean gains were evaluated for various body measurements and indices of lean tissue in barrows and gilts from 20 to 125 kg of BW. One genetic line was identified as the low-lean line [280 g of fat-free lean (FFL)/d], and the second line was the high-lean line (375 FFL gained/d). The experiment was conducted as a completely randomized design using a 2 x 2 x 5 factorial arrangement of treatments in 6 replicates (n = 120 pigs). The 2 genetic lines and sexes were provided ad libitum access to cornsoybean mixtures that met or exceeded their required amino acid requirements for their respective lean gain potentials. Six pigs of each sex and genetic line were slaughtered initially and at 25-kg of BW intervals to 125 kg of BW. Pigs slaughtered were measured for height, width, and length using metal calipers. Backfat and LM area were measured using real-time ultrasound, with backfat depth also measured using A-mode ultrasound technology. Longissimus muscle area and back-fat thickness at the 10th rib were measured on the chilled carcass. Data was analyzed using the MIXED procedure of SAS, with the animal as the experimental unit. Shoulders (P < 0.05) and lumbars (P < 0.05) were wider in the low-lean genetic line and in barrows. Gilts and the high-lean genetic line had less backfat and greater LM areas than the low-lean genetic line. As BW increased, there was a greater increase in FFL tissue and lower backfat depths in the high-lean vs. the low-lean genetic line. This resulted in a greater divergence of measurement values as BW increased. Femur weight, length, and cortical wall thickness were greater in the high-lean genetic line, but the differences were not significant. The high-lean genetic line had a greater (P < 0.01) organic matrix content in the femur and less ash, resulting in a lower percentage of bone ash (P < 0.01). The results indicate that differences occurred phenotypically between pigs having more muscle (wider hams) or more fat (wider shoulder and lumbar). As BW increased, the high-lean pigs had an increase in lean tissue, particularly after 75 kg of BW, and less backfat and less bone mineralization, whereas the low-lean line pigs had increased backfat and greater bone mineralization. Real-time ultrasound measurements using various formulas to estimate lean tissue produced values close to those determined from carcass measurements at 100 and 125 kg of BW. PMID- 17400971 TI - Effects of dietary supplementation with flax during prepuberty on fatty acid profile, mammogenesis, and bone resorption in gilts. AB - The possible role of dietary flax on pre-pubertal development of mammary glands and bone resorption was investigated in gilts. Fifty-seven gilts were fed 1 of 4 diets from 88 d of age until slaughter (d 212 +/- 1). Diets were control without flax (n = 14); 10% flaxseed supplementation (n = 13); 6.5% flaxseed meal supplementation (n = 15); and 3.5% flaxseed oil supplementation (n = 15). All diets were isonitrogenous and isocaloric. Jugular blood samples were obtained on d 78 and 210 to establish the fatty acid profile and to determine the concentrations of prolactin, estradiol, and cross-linked N-telopeptides of type I collagen. At slaughter, the mammary glands were excised, parenchymal and extraparenchymal tissues were dissected, and the composition of the parenchymal tissue (protein, fat, DM, and DNA) was determined. Histochemical analyses of the mammary parenchyma were performed, and fatty acid profiles in the extraparenchymal tissue were evaluated. Dietary flax increased (P < or = 0.001) the concentrations of PUFA and decreased those of SFA (P < 0.01) and MUFA (P < or = 0.001) in plasma and extraparenchymal tissues, which was largely due to the inclusion of 10% flaxseed or 3.5% flaxseed oil (P < or = 0.01) but not 6.5% flaxseed meal. Circulating concentrations of prolactin and estradiol were unaltered by treatments (P > 0.1), but concentrations of cross-linked N telopeptides of type I collagen tended to be greater (P < 0.1) in flax supplemented gilts. The DM content of parenchymal tissue was the only mammary compositional value affected, showing an increase with flax addition (P < 0.05). No change (P > or = 0.1) in the bromodeoxyuridine labeling index or estrogen receptor localization was observed with treatments. Dietary supplementation with flax as seed, meal, or oil, therefore, brought about the expected changes in the fatty acid profile but had no beneficial effects on mammary development or bone resorption. PMID- 17400972 TI - Preweaning housing effects on behavior and physiological measures in pigs during the suckling and fattening periods. AB - The effect of the preweaning housing system on the stress response of pigs before weaning and during fattening was studied in 33 litters of domestic pigs. Three preweaning housing systems were compared: barren crate (standard farrowing crate without straw), enriched crate (20% larger crate, with straw), and as a control, a farrowing pen (pen, 60% larger than the barren crate, with straw). At 25 d of age, pigs were tested with an isolation test and 1 d later with a human approach test (HumanT). Pigs were weaned at 28 d of age. At 3 and 6 mo of age, pigs were tested with an isolation-human approach test. The latency and frequency of squeal calls and locomotor activity were analyzed for all 3 tests, whereas physical contact with the human was also analyzed for the HumanT and isolation-human approach test. At 6 mo of age, the pigs were transported to a slaughterhouse. One day before transport, immediately after transport, and 1 h after transport, saliva samples were taken for cortisol analysis. The pH of the LM was also measured 45 min after slaughter. Preweaning housing system affected (P < 0.05) the probability of squeal vocalizations, the latency of locomotion, and the duration of locomotion during the HumanT. Pigs from the enriched pens vocalized less, had a longer latency to move, and performed less overall locomotion than pigs from the barren crates. Preweaning housing system did not affect behavior of fattening pigs. Cortisol concentrations before and after transport were not affected by preweaning housing system. An interaction of cortisol concentrations and housing systems was observed between the control sample and the sample taken immediately after transport in pigs from the barren crates (P < 0.05) compared with pigs from the enriched housing systems. Meat from pigs reared in the barren crate tended to have lower pH (P < 0.10) and that of pigs reared in enriched crates had lower pH (P < 0.05) than meat of pigs reared in enriched pens. No differences were observed between pigs from barren or enriched crates. Our results suggest that enrichment of the preweaning environment through enlarged space, provision of straw, and free movement for the sow had a positive effect on the coping behavior of pigs before weaning and prevented an increase in salivary cortisol concentration immediately after transport and a decrease in meat pH 45 min postmortem at the age of 6 mo. Minimal enrichment of the commercial farrowing crate did not affect behavior and physiological measures in pigs before and after weaning. PMID- 17400973 TI - Lectin binding profile of the small intestine of five-week-old pigs in response to the use of chlortetracycline as a growth promotant and under gnotobiotic conditions. AB - Antibiotics have traditionally been used for growth promotion in the pork industry; however, their use in animal feed has recently been limited because of human health concerns. The intestinal microbiota plays an important role in mediating many physiological functions such as digestion and animal growth. It was hypothesized that use of antibiotics as growth promotants and subsequent variations in intestinal microbiota induce significant changes in the intestinal glycoconjugate composition, which ultimately affects animal growth and disease susceptibility. The aim of this study was to characterize the lectin binding profiles of the ileum of weanling pigs in response to the absence of intestinal microbiota and to the use of the antibiotic chlortetracycline as growth promotant. Eighteen half-sib piglets obtained by cesarean section were divided into 3 treatment groups (n = 6) and maintained as control, antibiotic-fed, and gnotobiotic piglets until 5 wk of age. The glycoconjugate composition of the ileal tissues was examined by lectin histochemistry. Lycopersicon esculentum lectin, Jacalin, Pisum sativum agglutinin, Lens culinaris agglutinin (LCA), and Sambucus nigra lectin showed significant differences (P < 0.05) in binding intensities on the dome and villous epithelium between the treatment groups. Griffonia simplicifolia lectin I, Glycine maxi agglutinin, and Arachis hypogea agglutinin exhibited differences (P < 0.05) between treatment groups in lectin binding on goblet cells. Triticum vulgaris agglutinin, Pisum sativum agglutinin, and LCA showed significant differences (P < 0.05) in binding intensities on dome, corona, and follicular regions of the ileum among treatment groups of animals. Overall, ileal tissues from gnotobiotic piglets expressed significantly weaker (P < 0.05) lectin binding for many lectins compared with control and antibiotic groups. This suggests that the intestinal microbiota plays an important role in the expression of sugar moieties in the intestine. Lectins LCA, Phaseolus vulgaris Leucoagglutinin, and Maackia amurensis lectin II showed significant differences (P < 0.05) in lectin bindings between control and antibiotic-fed piglets. This indicates that chlortetracycline as a growth promotant induces biologically relevant changes in the lectin binding profile of the ileum. These findings will help in further understanding the role of the gut microbiota and the mechanisms of action of antibiotics as growth promotants in pigs. PMID- 17400974 TI - Daily dry matter intake to sustain body weight of mature, nonlactating, nonpregnant cows. AB - To quantify the relationship between DM consumption, the ability to sustain BW per unit of DMI (BW stasis), and days to reach BW equilibrium among diverse cattle breeds, weekly individual cow BW and DMI data were recorded for mature, nonpregnant, and nonlactating cows sampled from Angus, Braunvieh, Charolais, Hereford, Gelbvieh, Limousin, Pinzgauer, Red Poll, and Simmental breeds. Within each breed, cows were assigned to receive 1 of 4 daily DM allowances (56, 76, 93, or 111 g.BW(-0.75, kg)) of a ground alfalfa hay-corn grain-based diet. Cows were housed in pens (space for 4 animals/pen) in open-front barns and fed individually using head gates. During the first 60 d of the experiment, BW were recorded every 28 d, after which BW were recorded on a weekly basis until the cows were determined to have attained BW equilibrium. Individual cows were determined to be at BW equilibrium when the rate of weekly BW change did not differ from 0 over an 8-wk period. The number of days to reach BW equilibrium was not affected (P > 0.79) by breed but was affected by the daily DM allowance (P < 0.003). The number of days required to attain BW equilibrium was greater as the rate of feeding (g of DM fed.BW(-0.75)) increased and ranged from 103 to 136 d. Within breed linear and the pooled quadratic regressions were significant for BW. Observed breed differences varied with feeding rate. Weight stasis estimates for mature Red Poll cows (68.3 +/- 3.8) differed (P < 0.05) from estimates for all the breeds, with the exception of Limousin (72.0 +/- 3.8), Braunvieh (74.0 +/- 4.8), and Pinzgauer (75.5 +/- 3.8) cows at the lowest feeding rate. At the 111 g.BW(-0.75) daily DM allowance, the estimates for Limousin (82.2 +/- 3.8) were greater (P < 0.05) than for the other breeds, with the exception of the Pinzgauer (81.0 +/- 4.3) and Braunvieh (75.7 +/- 3.9), which were similar to the remaining breeds in the study (P > 0.05). The change in rank of breed estimates for BW stasis suggests a breed x nutrition interaction for BW stasis. PMID- 17400975 TI - Is complex regional pain syndrome an inflammatory process? Theories and therapeutic implications. PMID- 17400976 TI - Early oral analgesia after fast-track cardiac anesthesia. AB - PURPOSE: Oral analgesia after "fast-track" cardiac anesthesia has not been explored. The aim of this study was to compare two oral oxycodone analgesic regimens. METHODS: One hundred-twenty patients scheduled for coronary artery bypass grafting were randomly assigned postoperatively to receive immediate release oxycodone 5 mg and acetaminophen 325 mg (Percocet-5) (group I) per os four times daily, or controlled-release oxycodone 10 mg (OxyContin) (group II) per os every 12 hr and placebo twice daily. Acetaminophen 500 mg per os was used as first-line rescue medication, and immediate-release oxycodone (syrup form) 5 mg per os as second-line rescue medication. Pain intensity was assessed with a visual analogue scale on the first postoperative day, the morning after extubation, and thereafter four times daily for four days. Use of rescue medication and adverse events were recorded. RESULTS: Baseline demographic and operation-related characteristics were similar in both groups. While pain control was good in both groups, the immediate-release group experienced less pain on all postoperative days (P = 0.003), required significantly less rescue medication, and had fewer adverse effects such as somnolence and nausea. CONCLUSION: Peroral oxycodone is effective for early pain control after fast-track cardiac anesthesia. Immediate-release oxycodone/ acetaminophen appears to provide better analgesia and fewer side effects compared to controlled-release oxycodone. PMID- 17400977 TI - Role of amiodarone on the systemic inflammatory response induced by cardiac surgery: proinflammatory actions. AB - PURPOSE: Amiodarone (AMIO), a widely used anti-arrhythmic drug, has been shown to reduce the incidence of atrial fibrillation after cardiac surgery and also to exert immunomodulatory actions in vitro and proinflammatory effects in vivo. The present study investigated the immunomodulatory properties of AMIO in the inflammatory response induced by cardiac surgery with cardiopulmonary bypass (CPB). METHODS: In this double-blind, placebo-controlled trial, 20 patients undergoing elective coronary artery bypass graft were randomized to receive placebo or AMIO 600 mg day(-1) orally for seven days before surgery and 45 mg hr( 1) intravenously for 48 hr postoperatively. Plasma levels of the proinflammatory markers C-reactive protein (CRP), fibrinogen (FBG), tumour necrosis factor (TNF) alpha, interleukin (IL)-6 and monocyte chemoattractant protein (MCP)-1, and the antiinflammatory marker IL-10, were compared before and after surgery. RESULTS: Ninety-six hours after start of surgery, plasma levels of FBG had more than doubled (2.2 +/- 0.5-fold increase, P < 0.0001). Overall, FBG formation was significantly increased in the AMIO group (P = 0.048). Monocyte chemoattractant protein 1 secretion transiently increased four hours after start of surgery (6.6 +/- 4.5-fold increase) but rapidly declined thereafter, (P < 0.0001). There was a trend toward higher MCP-1 plasma concentrations in the AMIO group (P = 0.13). The plasma levels of CRP, TNF-alpha, IL-6 and Il-10 changed significantly over time, but were not altered by AMIO treatment. CONCLUSION: In the inflammatory response induced by cardiac surgery with CPB, our data suggest that AMIO treatment is associated with a selective trend toward proinflammatory actions. PMID- 17400978 TI - Anesthetic technique does not affect the performance of a rabbit model of arterial cyclic flow reductions: a pilot study. AB - PURPOSE: Pentobarbital anesthesia is, typically, used in an experimental model of cyclic flow reductions (CFR) in rabbits. Our initial observations, using a more complete and effective isoflurane-based anesthetic technique, failed to reproduce findings reported previously. Consequently, we compared the effects of these two anesthetic techniques in the model. METHODS: A modified Folts' model of carotid artery lesion and stenosis was used. Twelve rabbits completed the experimental protocol: five in the pentobarbital group (P) and seven in the isoflurane group (I). The carotid artery was exposed and flow was reduced by application of a clamp. A standardized injury was performed by cross clamping the artery with a needle forceps and this produced CFR. The number of CFR and the duration of their occurrence were noted. The incidence of thrombosis was compared in each group as well as hemodynamic, hematologic and bleeding time values. RESULTS: The hematocrit value, platelet count and bleeding time were similar in both groups. The median number and range of CFR [group P: 9 (4-16) ; group I: 9 (5-14)] and the time span of effective CFR formation (group P: 39 +/- 17; group I: 38 +/- 25 min) were comparable in both groups. The incidence of complete thrombosis of the carotid artery was similar in both groups. CONCLUSIONS: The stability of the model is of short duration, but the occurrence of CFR is not affected by the type of anesthesia. Our findings suggest that the ideal duration of the experimental protocol should be between 30 and 45 min in order to maximize the number of animals still developing CFR. PMID- 17400979 TI - Modified right-sided Broncho-Cath double lumen tube improves endobronchial positioning: a randomized study. AB - OBJECTIVE: A left-sided double lumen tube is recommended for one-lung ventilation for most thoracic surgeries, but for certain indications, a right-sided double lumen tube (R-DLT) may be mandatory. Frequent malposition of R-DLTs has been reported. We propose an innovative modification of Mallinckrodt's Broncho-Cath R DLT consisting of an enlarged area of the lateral orifice, and studied the impact of this modification on the adequacy of R-DLT positioning. METHODS: Eighty adult patients scheduled for elective thoracic surgery were randomized into two groups: standard Broncho-Cath R-DLT, or modified Broncho-Cath R-DLT. After induction of anesthesia, the R-DLT was positioned using a fibreoptic bronchoscope. The position of the R-DLT was assessed on three occasions: with the patient supine (T1), then immediately following the patient's transfer to the lateral position (T2), and after repositioning of the tube, when needed, with the patient in lateral position (T3). A score ranging from 1 to 4 was accorded to the relative position of the right upper lobe (RUL) orifice in relation to the origin of the RUL bronchus. RESULTS: The modified Broncho-Cath R-DLT was more frequently in an adequate position at T2: 77% vs 37% of patients (P = 0.0121), and easier to reposition at T3: 97% vs 74% of patients (P = 0.0109) in comparison to the standard Broncho-Cath R-DLT group. CONCLUSION: These data suggest the superiority of the modified Broncho-Cath R-DLT compared to a standard Broncho-Cath R-DLT for optimal R-DLT positioning to facilitate one-lung ventilation during thoracic surgery. PMID- 17400980 TI - Minimum effective anesthetic concentration (MEAC) for sciatic nerve block: subgluteus and popliteal approaches. AB - BACKGROUND: We tested the hypothesis that using a subgluteus approach to the sciatic nerve requires a lower concentration of mepivacaine to obtain complete anesthesia as compared with the popliteal approach. METHODS: With midazolam premedication (0.05 mg kg(-1) iv), 48 patients undergoing hallux valgus repair were randomly allocated to receive a sciatic nerve block using either a posterior popliteal (group Popliteal, n = 24) or subgluteus (group Subgluteus, n = 24) approach with 30 mL of local anesthetic injected after elicitation of plantar flexion of the foot with a current 30%) and ephedrine requirements were recorded. RESULTS: Forty-three patients in Group F and 42 patients in Group S completed the study. No differences in maximum sensory block height (Group F = median T2, interquartile range [T2-T4], Group S = T3 [T2-T4], P = 0.077) or time to achieve block height (F = 9.3 +/- 4.3 min, S = 9.7 +/- 4.7, P = 0.64) were observed. The frequencies of hypotension (Group F = 35/43, Group S = 32/42, P = 0.56), ephedrine utilization (Group F = 32/43, Group S = 26/42, P = 0.21) and nausea (Group F = 15/43, Group S = 16/42, P = 0.76) were similar. CONCLUSION: Rapid intrathecal injection of hyperbaric bupivacaine does not affect spread of spinal anesthesia or the incidence of hypotension and nausea in parturients. PMID- 17400982 TI - Case report: acute postoperative neurological impairment from fat embolism syndrome. AB - PURPOSE: To describe a case of fat embolism syndrome (FES) following elective tendon contracture release in a patient with myotonic dystrophy, to highlight the importance of considering this entity in the differential diagnosis of acute postoperative neurocognitive dysfunction. CLINICAL FEATURES: A 34-yr-old man with myotonic dystrophy underwent uneventful tendon contracture release under regional anesthesia. In the immediate postoperative period, neurological and respiratory complications developed, requiring intensive care support. The patient showed the classical clinical triad of hypoxemia, neurological impairment and a petechial rash associated with the FES. A diagnosis of FES was made and, despite therapy including fluid and inotropic support, the patient succumbed to the condition. There was no demonstrated intracardiac shunt, suggesting a physiological intrapulmonary shunt was responsible for the development of systemic manifestations of FES. CONCLUSIONS: Postoperative neurological dysfunction is a difficult condition with numerous possible causes. All possible etiologies, including FES, need to be considered in the differential diagnosis and postoperative management of patients developing acute postoperative neurological impairment and hypoxemia. PMID- 17400983 TI - Case report: fentanyl-associated intraoperative anaphylaxis with pulmonary edema. AB - PURPOSE: To describe an atypical presentation of intraoperative anaphylaxis due to fentanyl. CLINICAL FEATURES: A 40-yr-old otherwise healthy woman was admitted for abdominal hysterectomy. She denied any drug allergies or past adverse anesthetic reactions. Physical examination, vital signs, and laboratory findings were all within normal limits. Twenty minutes after induction of general anesthesia with propofol, lidocaine, fentanyl, and rocuronium, she developed sudden onset of hypotension and bronchospasm. She was treated with fluids and epinephrine, but nonetheless required mechanical ventilation for 48 hr. Chest x ray revealed pulmonary edema which resolved over two days. She recovered completely and was discharged home. Subsequent skin testing showed reactions to fentanyl and succinylcholine. Because the patient had not received succinylcholine, the cause of her anaphylaxis was attributed to fentanyl. The patient later returned for her hysterectomy and tolerated spinal anesthesia with bupivacaine and morphine. CONCLUSION: Anaphylaxis is a fulminant, unexpected, IgE mediated allergic reaction which can be triggered by multiple agents. Common causative agents include neuromuscular blocking drugs, latex, antibiotics, colloids, hypnotics, and opioids. Fentanyl, however, is an extremely unusual cause of anaphylaxis. Pulmonary edema, although uncommon in anaphylaxis, can be a prominent feature, as was the case with this patient. PMID- 17400984 TI - Case series: the McGrath videolaryngoscope--an initial clinical evaluation. AB - PURPOSE: To document tracheal intubation success rates and airway instrumentation times using the newly designed McGrath videolaryngoscope. METHODS: We prospectively recorded factors associated with difficult tracheal intubation, factors causing actual difficulty in tracheal intubation, as well as complications arising from use of the new McGrath videolaryngoscope in a series of adult patients with normal preoperative airway examinations. All patients were undergoing scheduled or elective surgery. In the first 75 patients (phase I), experience with airway instrumentation was documented, while in the second 75 patients (phase II), the time required to obtain an optimal view of the larynx was recorded, as well as the time to complete tracheal intubation. RESULTS: Ninety-eight percent of all tracheal intubations were successful using the McGrath videolaryngoscope. Cormack and Lehane grade I views were obtained in 143 patients (95%) and grade II views were achieved in six (4%). In phase II, the median time required to obtain an adequate view was 6.3 sec [interquartile range 4.7-8.7 (range 2-26.3)], and to complete tracheal intubation was 24.7 sec [18.5 34.4 (11.4-286)]. Fortynine (65%) of the tracheal intubations were completed within 30 sec, and 72 (96%) were completed within one minute. No complications were encountered in any patient. CONCLUSIONS: The McGrath videolaryngoscope is an effective aid to airway management in patients with normal airways, based upon intubation success rates and the ability to rapidly secure the airway. Its potential advantages of convenience and portability warrant further evaluation in comparison with other airway devices and in patients with difficult airways. PMID- 17400985 TI - Research awards program of the Canadian Anesthesiologists' Society/Canadian Anesthesia Research Foundation: survey of past recipients. AB - PURPOSE: To review the effectiveness of the Research Grants, Career Scientist Award and Fellowship Program (RGCSFP) of the Canadian Anesthesiologists' Society (CAS)/Canadian Anesthesia Research Foundation (CARF) by surveying past recipients. METHODS: The CAS Research Committee database of past RGCSFP recipients was reviewed to identify all past award recipients from the time of program inception in 1985 until the year 2005. A questionnaire was mailed to all past recipients with retrievable addresses. The questionnaire asked specific questions regarding past and current research, publications, grant procurement and mentoring of trainees. In addition, opinions regarding the program's effectiveness were solicited. RESULTS: Of the 100 mailed survey forms, 66 (66%) were completed and returned. The number of original published articles per respondent was 30.1 +/- 28.5 (mean +/- SD) at the time of the survey, and the life-time number of all publication types was 38.5 +/- 34.6. Cumulative research funding increased in relation to the number of years in research, and amongst past recipients with > 15 years in research, individual research funding from all sources was $585,747 +/- $773,716. Ninety-six percent of respondents agreed or strongly agreed that the program was valuable. CONCLUSIONS: The RGCSFP has supported a large number of new and established anesthesia investigators in Canada, and these individuals have demonstrated high research productivity. The program is considered to be very valuable according to past recipients. PMID- 17400986 TI - Beta-blockers and anesthetic preconditioning: friend or foe? PMID- 17400987 TI - Use of the "Aretube" to facilitate ventilation during percutaneous tracheostomy. PMID- 17400988 TI - Another complication associated with videolaryngoscopy. PMID- 17400989 TI - Failure of an iv fluid warming device. PMID- 17400990 TI - Regional anesthesia for a patient with hereditary neuropathy with liability to pressure palsies. PMID- 17400991 TI - A case of paradoxical cerebral thromboembolism after removal of a central venous catheter. PMID- 17400992 TI - Food and Drug Administration public hearing of the conduct of emergency clinical research: testimony of Dr. Dutton. PMID- 17400993 TI - Recommendations for implementation of community consultation and public disclosure under the Food and Drug Administration "exception from informed consent requirements for emergency research": testimony of the American Heart Association. PMID- 17400994 TI - Response to Food and Drug Administration draft guidance statement on research into the treatment of life-threatening emergency conditions using exception from informed consent: testimony of the Resuscitation Outcomes Consortium. PMID- 17400995 TI - Ultrasound use in the diagnosis of abdominal aortic aneurysm. PMID- 17400996 TI - The effect of emergency department expansion on emergency department overcrowding. AB - OBJECTIVES: To examine the effects of emergency department (ED) expansion on ambulance diversion at an urban, academic Level 1 trauma center. METHODS: This was a pre-post study performed using administrative data from the ED and hospital electronic information systems. On April 19, 2005, the adult ED expanded from 28 to 53 licensed beds. Data from a five-month pre-expansion period (November 1, 2004, to March 1, 2005) and a five-month postexpansion period (June 1, 2005, to October 31, 2005) were included for this analysis. ED and waiting room statistics as well as diversion status were obtained. Total ED length of stay (LOS) was defined as the time from patient registration to the time leaving the ED. Admission hold LOS was defined as the time from the inpatient bed request to the time leaving the ED for admitted patients. Mean differences (95% confidence interval [CI]) in total time spent on ambulance diversion per month, diversion episodes per month, and duration per diversion episode were calculated. An accelerated failure time model was performed to test if ED expansion was associated with a reduction in ambulance diversion while adjusting for potential confounders. RESULTS: From pre-expansion to postexpansion, daily patient volume increased but ED occupancy decreased. There was no significant change in the time spent on ambulance diversion per month (mean difference, 10.9 hours; 95% CI = 74.0 to 95.8), ambulance diversion episodes per month (two episodes per month; 95% CI = -4.2 to 8.2), and duration of ambulance diversion per episode (0.3 hours; 95% CI = -4.0 to 3.5). Mean (+/-SD) total LOS increased from 4.6 (+/-1.9) to 5.6(+/-2.3) hours, and mean (+/-SD) admission hold LOS also increased from 3.0 (+/-0.2) to 4.1 (+/-0.2) hours. The proportion of patients who left without being seen was 3.5% and 2.7% (p = 0.06) in the pre-expansion and postexpansion periods, respectively. In the accelerated failure time model, ED expansion did not affect the time to the next ambulance diversion episode. CONCLUSIONS: An increase in ED bed capacity did not affect ambulance diversion. Instead, total and admission hold LOS increased. As a result, ED expansion appears to be an insufficient solution to improve diversion without addressing other bottlenecks in the hospital. PMID- 17400997 TI - Struggling with the rule: the exception from informed consent in resuscitation research. PMID- 17400998 TI - The research on community consultation: an annotated bibliography. AB - Community consultation is a required element of research studies that use a waiver of or exception from informed consent. Its intent is to provide an additional patient safeguard in emergency research circumstances when prospective informed consent is not possible. Investigators have reported that community consultation may be the most difficult aspect in implementing research trials using a wavier of or exception from informed consent. This article presents a brief overview of the sparse literature available on the process of community consultation since the inception of the current emergency research regulations. To determine if the process is meeting its goals, more research will be required. PMID- 17400999 TI - Emergency department categorization. PMID- 17401000 TI - Moving closer to an operational definition for ED crowding. PMID- 17401002 TI - Surgery after primary chemoradiotherapy in squamous cancer of the esophagus: is the photon mightier than the sword? PMID- 17401003 TI - Gynecologic oncology or medical oncology: what's in a name? PMID- 17401004 TI - Chemoradiation followed by surgery compared with chemoradiation alone in squamous cancer of the esophagus: FFCD 9102. AB - PURPOSE: Uncontrolled studies suggest that chemoradiation has similar efficacy as surgery for esophageal cancer. Therefore, a randomized trial was carried out to compare, in responders only, chemoradiation alone with chemoradiation followed by surgery in patients with locally advanced tumors. PATIENTS AND METHODS: Eligible patients had operable T3N0-1M0 thoracic esophageal cancer. Patients received two cycles of fluorouracil (FU) and cisplatin (days 1 to 5 and 22 to 26) and either conventional (46 Gy in 4.5 weeks) or split-course (15 Gy, days 1 to 5 and 22 to 26) concomitant radiotherapy. Patients with response and no contraindication to either treatment were randomly assigned to surgery (arm A) or continuation of chemoradiation (arm B; three cycles of FU/cisplatin and either conventional [20 Gy] or split-course [15 Gy] radiotherapy). Chemoradiation was considered equivalent to surgery if the difference in 2-year survival rate was less than 10%. RESULTS: Of 444 eligible patients, 259 were randomly assigned; 230 patients (88.8%) had epidermoid cancer, and 29 (11.2%) had glandular carcinoma. Two-year survival rate was 34% in arm A versus 40% in arm B (hazard ratio for arm B v arm A = 0.90; adjusted P = .44). Median survival time was 17.7 months in arm A compared with 19.3 months in arm B. Two-year local control rate was 66.4% in arm A compared with 57.0% in arm B, and stents were less required in the surgery arm (5% in arm A v 32% in arm B; P < .001). The 3-month mortality rate was 9.3% in arm A compared with 0.8% in arm B (P = .002). Cumulative hospital stay was 68 days in arm A compared with 52 days in arm B (P = .02). CONCLUSION: Our data suggest that, in patients with locally advanced thoracic esophageal cancers, especially epidermoid, who respond to chemoradiation, there is no benefit for the addition of surgery after chemoradiation compared with the continuation of additional chemoradiation. PMID- 17401005 TI - Does ovarian cancer treatment and survival differ by the specialty providing chemotherapy? AB - PURPOSE: Chemotherapy for ovarian cancer is usually administered by medical oncologists (MOs) or gynecologic oncologists (GOs). GOs perform a broad spectrum of surgical and medical activities while managing a limited number of diseases; MOs specialize in the administration of chemotherapy but manage a broad array of diseases. We asked whether survival, treatment, and toxicity differed according to the type of specialist providing the chemotherapy after surgery. PATIENTS AND METHODS: Using Surveillance, Epidemiology, and End Results (SEER)--Medicare data for patients 65 years old from 1991 through 2001 from eight SEER sites, we identified 344 patients with ovarian cancer who were treated with chemotherapy by a GO after surgery. Using optimal matching and propensity scores based on 36 characteristics, we matched these patients to 344 similar patients who were operated on and staged by the same type of surgeon but who received chemotherapy from an MO. RESULTS: MOs administered chemotherapy over more weeks than did the GOs (16.5 v 12.1 weeks, respectively; P < .0023), and MO patients had substantially more weeks that included chemotherapy-associated adverse events than GO patients (16.2 v 8.9 weeks, respectively; P < .0001). However, there was no difference in 5-year survival rate between the GO and MO groups (35% v 34%, respectively; P = .45). CONCLUSION: GO- and MO-treated patients who were closely matched on prognostic characteristics experienced very different rates of chemotherapy-associated adverse events and very different chemotherapy treatment styles by specialty type; however, their survival was virtually identical. PMID- 17401006 TI - Extraosseous localized ewing tumors: improved outcome with anthracyclines--the French society of pediatric oncology and international society of pediatric oncology. AB - PURPOSE: To evaluate the outcome of children with an extraosseous Ewing tumor (EOE) according to treatment. PATIENTS AND METHODS: Children with EOE were treated either with the strategy used for malignant mesenchymal tumors (MMTs) by the International Society of Pediatric Oncology (SIOP) or with the French Society of Pediatric Oncology (SFOP) regimen used for osseous Ewing tumors (OET). The MMT strategy included vincristine/actinomycin for small and resected tumors or ifosfamide/vincristine/actinomycin for unfavorable sites or unresectable tumors. Surgical excision was to be attempted after four courses, followed by local irradiation in case of residue. Osseous Ewing (OE) protocol included three courses of cyclophosphamide/doxorubicin followed either by two similar courses in case of good response or two courses of ifosfamide/etoposide in case of no response. After resection of the primary, treatment included conventional chemotherapy in case of good histologic response and high-dose chemotherapy and radiotherapy for poor response. All diagnosis specimens were reviewed by the panel. RESULTS: Between 1989 and 1999, 63 patients were registered. Characteristics of patients treated by both protocols were similar. Five-year overall survival (OS) and event-free survival (EFS) of those treated with the OE protocol are 83% and 75%, respectively, which is significantly better than the OS and EFS of those treated with the MMT strategy (59% and 44%, respectively; P = .04 and .008, respectively). The size of the primary and the type of protocol influenced patients' EFS. In multivariate analysis, only the regimen had an impact on OS and EFS. CONCLUSION: Our study shows that patients with EOE should be treated with OE regimens, probably because of the use of anthracyclines. PMID- 17401007 TI - Obesity in survivors of childhood acute lymphoblastic leukemia and lymphoma. AB - PURPOSE: We evaluated the long-term effects of treatment on the body mass index (BMI) of children with acute lymphoblastic leukemia (ALL) or lymphoblastic lymphoma who received one of three CNS-directed therapies: intrathecal methotrexate with intravenous high-dose methotrexate (1 g/m2), intrathecal methotrexate with 18 Gy cranial radiation, or intrathecal methotrexate with 24 Gy cranial radiation. PATIENTS AND METHODS: Between 1979 and 1984, 456 children with newly diagnosed ALL and lymphoma were enrolled onto a single protocol at St Jude Children's Research Hospital (Memphis, TN). The heights and weights of 422 of the children were measured at diagnosis, during treatment, at the end of therapy, and approximately every 6 to 12 months thereafter. Patients who had attained their adult height at the time of analysis (n = 248) were placed in weight categories based on their BMI, BMI percentile, or weight-for-length percentile depending on age. RESULTS: The overall percentage of survivors who were overweight or obese approximated rates prevalent in the general population of the United States. Young age (< 6 years) and overweight/obesity at diagnosis were the best predictors of obesity at adult height. The rate of BMI increase did not differ significantly between children who received radiation and those who did not, nor between patients who received 18 or 24 Gy of cranial radiation. CONCLUSION: BMI weight category at diagnosis, rather than type of CNS treatment received, predicted adult weight in long-term survivors of childhood hematologic malignancies. PMID- 17401008 TI - Early changes in auditory function as a result of platinum chemotherapy: use of extended high-frequency audiometry and evoked distortion product otoacoustic emissions. AB - PURPOSE: The objective is to describe progressive changes in hearing and cochlear function in children and adolescents treated with platinum-based chemotherapy and to begin preliminary evaluation of the feasibility of extended high-frequency audiometry and distortion product otoacoustic emissions for ototoxicity monitoring in children. PATIENTS AND METHODS: Baseline and serial measurement of conventional pure-tone audiometry (0.5 to 8 kHz) and evoked distortion product otoacoustic emissions (DPOAEs) were conducted for 32 patients age 8 months to 20 years who were treated with cisplatin and/or carboplatin chemotherapy. Seventeen children also had baseline and serial measurement of extended high-frequency (EHF) audiometry (9 to 16 kHz). Audiologic data were analyzed to determine the incidence of ototoxicity using the American Speech-Language-Hearing Association criteria, and the relationships between the different measures of ototoxicity. RESULTS: Of the 32 children, 20 (62.5%) acquired bilateral ototoxicity in the conventional frequency range during chemotherapy treatment, and 26 (81.3%) had bilateral decreases in DPOAE amplitudes and dynamic range. Of the 17 children with EHF audiometry results, 16 (94.1%) had bilateral ototoxicity in the EHF range. Pilot data suggest that EHF thresholds and DPOAEs show ototoxic changes before hearing loss is detected by conventional audiometry. CONCLUSION: EHF audiometry and DPOAEs have the potential to reveal earlier changes in auditory function than conventional frequency audiometry during platinum chemotherapy in children. PMID- 17401009 TI - Molecular profiling identifies prognostic subgroups of pediatric glioblastoma and shows increased YB-1 expression in tumors. AB - PURPOSE: Pediatric glioblastoma (pGBM) is a rare, but devastating brain tumor. In contrast to GBM in adults (aGBM), little is known about the mechanisms underlying its development. Our aim is to gain insight into the molecular pathways of pGBM. MATERIALS AND METHODS: Thirty-two pGBM and seven aGBM samples were investigated using biochemical and transcriptional profiling. Ras and Akt pathway activation was assessed through the phosphorylation of downstream effectors, and gene expression profiles were generated using the University Health Network Human 19K cDNA arrays. Results were validated using real-time polymerase chain reaction and immunohistochemistry and compared with existing data sets on aGBM. RESULTS: There are at least two subsets of pGBM. One subset, associated with Ras and Akt pathway activation, has very poor prognosis and exhibits increased expression of genes related to proliferation and to a neural stem-cell phenotype, similar to findings in aggressive aGBM. This subset was still molecularly distinguishable from aGBM after unsupervised and supervised analysis of expression profiles. A second subset, with better prognosis, is not associated with activation of Akt and Ras pathways, may originate from astroglial progenitors, and does not express gene signatures and markers shown to be associated with long-term survival in aGBM. Both subsets of pGBM show overexpression of Y-box-protein-1 that may help drive oncogenesis in this tumor. CONCLUSION: Our work, the first study of gene expression profiles in pGBM, provides valuable insight into active pathways and targets in a cancer with minimal survival, and suggests that these tumors cannot be understood exclusively through studies of aGBM. PMID- 17401010 TI - Activated intrinsic apoptosis pathway is a key related prognostic parameter in acute myeloid leukemia. AB - PURPOSE: By parallel assessment of multiple apoptosis-related transcripts, we aimed to refine the current concept of apoptosis resistance in acute myeloid leukemia (AML) and identify the combination of genes best predicting overall survival (OS). PATIENTS AND METHODS: The reverse transcriptase multiplex ligation dependent probe amplification technique was used for simultaneous quantification of 31 apoptosis-related transcripts in viable (7AAD-/AnnexinV-) blasts (CD45dim) from bone marrow aspirates of 120 newly diagnosed AML patients. By forward selection, a prognosis-predicting gene expression profile was constructed. The predictive validity of this profile was assessed by cross validation. RESULTS: High transcript levels were associated with poor OS for seven of 31 genes, three of which were proapoptotic. The average expression of all 12 antiapoptotic genes was associated with poor OS (P = .029). A similar association with poor OS was found for the average expression of all 19 proapoptotic genes (P = .009). Forward selection and cross validation revealed the antiapoptotic gene BIRC3 and the proapoptotic genes BAX-(l) and BMF to optimally predict OS. Three equally sized patient groups, constructed by ranking the cross-validated prognoses of the patients, were clearly distinct (median OS times were 8.2, 16.7, and 85.6 months). CONCLUSION: High expression of both pro- and antiapoptotic genes predicted poor OS, which postulates a mechanism of activation of the apoptosis pathway as a whole. This mechanism, which culminates in a three-gene expression signature, allows accurate clinical outcome prediction in AML and puts efforts to target single antiapoptosis genes in a new perspective. PMID- 17401011 TI - Impact of delirium on cognition, distress, and health-related quality of life after hematopoietic stem-cell transplantation. AB - PURPOSE: To determine the impact of delirium during the acute phase of myeloablative hematopoietic stem-cell transplantation (HSCT) on health-related quality of life (HRQOL), distress, and neurocognitive functioning 30 and 80 days after transplantation. PATIENTS AND METHODS: Ninety patients completed a battery assessing HRQOL, distress, and neuropsychological functioning before receiving their first HSCT. Delirium was assessed three times per week using the Delirium Rating Scale and the Memorial Delirium Assessment Scale from 7 days before transplantation through 30 days after transplantation. At 30 days after transplantation, distress and neurocognitive functioning were assessed. At 80 days after transplantation, HRQOL, distress, and neuropsychological functioning were re-evaluated. RESULTS: After adjusting for confounding factors, patients who experienced a delirium episode, versus patients who did not, reported significantly worse depression, anxiety, and fatigue symptoms at 30 days (linear regression beta(s) = 0.2, 0.3, and 0.5, respectively; P < .04). At 80 days, patients with a delirium episode had significantly worse executive functioning (beta = -1.1; P < .02), attention and processing speed (beta(s) = -4.7 and -5.4, respectively; P < .03), mental health on the Medical Outcomes Study Health Survey, 12-item short form (beta = -6.5; P < .02), and anxiety, fatigue, and cancer and treatment distress symptoms (beta(s) = 0.4, 0.6, and 0.3, respectively; P < .03). CONCLUSION: Patients with a malignancy who experience delirium during myeloablative HSCT showed impaired neurocognitive abilities and persistent distress 80 days after transplantation. Effective prevention or treatment of delirium during HSCT may improve both cognitive and psychological outcomes. PMID- 17401012 TI - Definition of clinically distinct molecular subtypes in estrogen receptor positive breast carcinomas through genomic grade. AB - PURPOSE: A number of microarray studies have reported distinct molecular profiles of breast cancers (BC), such as basal-like, ErbB2-like, and two to three luminal like subtypes. These were associated with different clinical outcomes. However, although the basal and the ErbB2 subtypes are repeatedly recognized, identification of estrogen receptor (ER) -positive subtypes has been inconsistent. Therefore, refinement of their molecular definition is needed. MATERIALS AND METHODS: We have previously reported a gene expression grade index (GGI), which defines histologic grade based on gene expression profiles. Using this algorithm, we assigned ER-positive BC to either high-or low-genomic grade subgroups and compared these with previously reported ER-positive molecular classifications. As further validation, we classified 666 ER-positive samples into subtypes and assessed their clinical outcome. RESULTS: Two ER-positive molecular subgroups (high and low genomic grade) could be defined using the GGI. Despite tracking a single biologic pathway, these were highly comparable to the previously described luminal A and B classification and significantly correlated to the risk groups produced using the 21-gene recurrence score. The two subtypes were associated with statistically distinct clinical outcome in both systemically untreated and tamoxifen-treated populations. CONCLUSION: The use of genomic grade can identify two clinically distinct ER-positive molecular subtypes in a simple and highly reproducible manner across multiple data sets. This study emphasizes the important role of proliferation-related genes in predicting prognosis in ER positive BC. PMID- 17401013 TI - Pharmacogenetic profiling in patients with advanced colorectal cancer treated with first-line FOLFOX-4 chemotherapy. AB - PURPOSE: The objective is to investigate whether polymorphisms with putative influence on fluorouracil/oxaliplatin activity are associated with clinical outcomes of patients with advanced colorectal cancer treated with first-line oxaliplatin, folinic acid, and fluorouracil palliative chemotherapy. MATERIALS AND METHODS: Consecutive patients were prospectively enrolled onto medical oncology units in Central Italy. Patients were required to have cytologically/histologically confirmed metastatic disease with at least one measurable lesion. Peripheral blood samples were used for genotyping 12 polymorphisms in thymidylate synthase, methylenetetrahydrofolate reductase, xeroderma pigmentosum group D (XPD), excision repair cross complementing group 1 (ERCC1), x-ray cross complementing group 1, x-ray cross complementing protein 3, glutathione S-transferases (GSTs) genes. The primary end point of the study was to investigate the association between genotypes and progression-free survival (PFS). RESULTS: In 166 patients, ERCC1-118 T/T, XPD-751 A/C, and XPD-751 C/C genotypes were independently associated with adverse PFS. The presence of two risk genotypes (ERCC1-118 T/T combined with either XPD-751 A/C or XPD-751 C/C) occurred in 50 patients (31%). This profiling showed an independent role for unfavorable PFS with a hazard ratio of 2.84% and 95% CI of 1.47 to 5.45 (P = .002). Neurotoxicity was significantly associated with GSTP1-105 A/G. Carriers of the GSTP1-105 G/G genotype were more prone to suffer from grade 3 neurotoxicity than carriers of GSTP1-105 A/G and GSTP1-105 A/A genotypes. CONCLUSION: A pharmacogenetic approach may be an innovative strategy for optimizing palliative chemotherapy in patients with advanced colorectal cancer. These findings deserve confirmation in additional prospective studies. PMID- 17401014 TI - Patient assessment of bowel function during and after pelvic radiotherapy: results of a prospective phase III North Central Cancer Treatment Group clinical trial. AB - PURPOSE: To evaluate patient assessment of long-term effect of pelvic radiotherapy on bowel function. PATIENTS AND METHODS: As part of a prospective randomized trial evaluating the impact of sucralfate on bowel function in patients receiving pelvic radiotherapy, patient-assessed bowel function data were collected during radiotherapy and again at 4 weeks and 1 year after completion of radiotherapy. RESULTS: The number of bowel movements per day increased up to week 4 and then slowly decreased. At 1 year, the mean number of bowel movements per day had increased from 1.75 to 2.09 and the median from 1 to 2. All measures of adverse bowel function worsened during radiotherapy. Frequency of bowel movements and symptoms of frequency, nocturnal bowel movements, cramping, and bleeding returned close to baseline values by 1 year. Symptoms of urgency, clustering, and measures of incontinence were all persistent at 1 year. The mean increase in bowel function score at 1 year was 0.74 (range, -5 to 7). CONCLUSION: Pelvic radiotherapy is associated with a slight increase in bowel movement frequency and decrease in several patient-reported measures of adverse bowel function. Several measures of adverse function persisted at 1 year. PMID- 17401015 TI - Regression after whole-brain radiation therapy for brain metastases correlates with survival and improved neurocognitive function. AB - PURPOSE: Brain metastasis (BM) is a major cause of suffering and health costs in cancer patients. Whole-brain radiation therapy (WBRT) offers tumor shrinking and palliation in many cases, but it has been speculated that these benefits may be outweighed by adverse effects on neurocognitive function (NCF). PATIENTS AND METHODS: Two hundred eight BM patients from the WBRT arm of phase III trial PCI P120-9801 evaluating motexafin gadolinium were analyzed. NCF, assessed by tests of memory, executive function, and fine motor coordination, was correlated to magnetic resonance imaging-measured BM volume. NCF and survival were compared in 135 patients assessable at 2 months with tumor shrinkage below (poor responders) and above (good responders) the population median (45%). Mean NCF scores and BM volume at 4 and 15 months were compared. RESULTS: Good responders experienced a significantly improved survival (unidirectional P = .03). For all tests, the median time to NCF deterioration was longer in good compared with poor responders, with statistical significance seen for Trailmaking B (executive function), and two Pegboard tests (fine motor). In long-term survivors, tumor shrinkage significantly correlated with preservation of executive function and fine motor coordination (r = 0.68 to 0.88). During the early follow-up period, the population mean NCF scores were dominated by patients with progressive disease. A small subset of 15-month survivors had stable or improving scores, and greater mean BM reduction. CONCLUSION: WBRT-induced tumor shrinkage correlates with better survival and NCF preservation. NCF is stable or improved in long-term survivors. Tumor progression adversely affects NCF more than WBRT does, thus making enhancement of radiation response a worthwhile aim in this patient population. PMID- 17401016 TI - Direct-to-consumer advertising in oncology: a content analysis of print media. AB - PURPOSE: Content analysis of cancer-related direct-to-consumer advertising (DTCA), with a focus on how benefit and risk/adverse effect information is presented, is essential to understanding its potential impact on oncology outcomes. METHODS: We reviewed all oncology DTCA appearing in three patient focused cancer magazines and a sample of selected popular magazines from January 2003 to June 2006. We determined the Flesch reading ease score (FRES) for the text in each advertisement (a score > or = 65 is readable for the average person). We also assessed the proportion, type size, and placement of benefits and risks/adverse effects, as well as the nature and content of advertising appeals. RESULTS: Of 284 advertisements identified, 49 were unique. Oncology related DTCA was rare in the popular magazines, and appeared mostly in those aimed at female readership. About equal amounts of text were devoted to benefits and risks/adverse effects, and all text was difficult to read. The mean FRES for benefit text was 39.71; for risk/adverse effect text, it was 38.22, a difference of 1.49 (95% CI, -4.02 to 7.00). The largest font size for benefits was 4.60 mm on average; for risks/adverse effects, it was 2.38 mm, a difference of 2.22 mm (95% CI, 1.35 to 3.09). Appeals to medication effectiveness were frequent (95%) and often made with clinical trial data (61%). CONCLUSION: Oncology print DTCA is prevalent in cancer-related, patient-directed magazines, and infrequent in the popular press. The information presented is considerably difficult to read, raising important questions about the appropriateness of direct-to-consumer marketing for oncologic medications. PMID- 17401017 TI - Treatment tolerance and efficacy in geriatric oncology: a systematic review of phase III randomized trials conducted by five National Cancer Institute-sponsored cooperative groups. AB - PURPOSE: Elderly patients share the majority of the disease burden in cancer. Although 61% of new cancer cases occur among elderly, they comprise only 25% of the patients enrolled onto randomized clinical trials (RCTs). A systematic review to assess the accurate participation of elderly patients in RCTs has not been performed. PATIENTS AND METHODS: We reviewed all consecutively completed phase III RCTs conducted by five National Cancer Institute-sponsored cooperative groups. Published papers and study protocols were used for data extraction. We used a cutoff age of 65 years to define elderly patients. For trials that did not exclusively enroll elderly, data were extracted on number of participants 65 years of age. Outcome between the innovative and the standard treatment was compared. RESULTS: Of 345 studies, only one trial exclusively enrolled elderly patients (0.28%); 57% of the trials (n = 197) had no stratification by age, and 12% of the studies had a stratification age 65 years (n = 42). Overall survival in the trial exclusively enrolling elderly favored the newer treatments (hazard ratio [HR], 0.69; 95% CI, 0.47 to 1.02; P = .06). Additionally, in trials enrolling more than 40% of elderly, survival and event-free survival favored the innovative treatments (HR for survival, 0.91; 95% CI, 0.84 to 0.99; P = .03; HR for event-free survival, 0.85; 95% CI, 0.72 to 1.01; P = .07). Treatment-related mortality was similar in both the innovative and standard treatment groups (HR, 0.91; 95% CI, 0.47 to 1.78; P = .8). CONCLUSION: Our data indicate that enrollment of elderly in experimental RCTs is not associated with increased harm to this patient population. Increased participation of elderly may help in finding new treatments that are clinically applicable specifically to this cohort of patients. PMID- 17401018 TI - Left atrial sarcoma mimicking obstructive pulmonary disease. PMID- 17401019 TI - Intravascular large cell lymphoma of the natural killer cell type. PMID- 17401020 TI - Successful chemotherapy for advanced nonseminomatous germ-cell tumor in a patient undergoing chronic hemodialysis. PMID- 17401021 TI - Wine for appetite loss: "how do you know?". PMID- 17401022 TI - CT2--the clinical trials control tower: overcoming barriers to opening oncology clinical trials. PMID- 17401023 TI - CD57 expression on lymphoma microenvironment as a new prognostic marker related to immune dysfunction. PMID- 17401024 TI - Is prophylactic cranial irradiation a possible option for human epidermal growth factor receptor 2-positive breast cancer? PMID- 17401025 TI - Carcinoembryonic antigen monitoring for early detection of asymptomatic incurable metastatic colorectal cancer. PMID- 17401026 TI - Potential chemotherapy options in the triple negative subtype of breast cancer. PMID- 17401027 TI - Bone marrow and not bone is the primary site for skeletal metastasis: critical role of [18F]fluorodeoxyglucose positron emission tomography in this setting. PMID- 17401028 TI - Control arm for surgery alone is needed but difficult to obtain in randomized trials for adjuvant chemotherapy after liver resection for colorectal metastases. PMID- 17401029 TI - Effectiveness after 1 year of a short-term physical activity intervention on cardiorespiratory fitness in cancer patients. PMID- 17401030 TI - Neuropeptidomics strategies for specific and sensitive identification of endogenous peptides. AB - A new approach using targeted sequence collections has been developed for identifying endogenous peptides. This approach enables a fast, specific, and sensitive identification of endogenous peptides. Three different sequence collections were constituted in this study to mimic the peptidomic samples: SwePep precursors, SwePep peptides, and SwePep predicted. The searches for neuropeptides performed against these three sequence collections were compared with searches performed against the entire mouse proteome, which is commonly used to identify neuropeptides. These four sequence collections were searched with both Mascot and X! Tandem. Evaluation of the sequence collections was achieved using a set of manually identified and previously verified peptides. By using the three new sequence collections, which more accurately mimic the sample, 3 times as many peptides were significantly identified, with a false-positive rate below 1%, in comparison with the mouse proteome. The new sequence collections were also used to identify previously uncharacterized peptides from brain tissue; 27 previously uncharacterized peptides and potentially bioactive neuropeptides were identified. These novel peptides are cleaved from the peptide precursors at sites that are characteristic for prohormone convertases, and some of them have post translational modifications that are characteristic for neuropeptides. The targeted protein sequence collections for different species are publicly available for download from SwePep. PMID- 17401031 TI - Can deficits in social problem-solving in people with personality disorder be reversed? AB - Research evidence is beginning to emerge that social problem-solving can improve the social functioning of people with personality disorder. This approach is particularly important because it may be relatively easy to train healthcare workers to deliver this intervention. However, the costs and cost-effectiveness of social problem-solving need to be established if it is to be made more widely available. PMID- 17401032 TI - Management of insomnia in older adults. AB - Complaints of insomnia are very common, especially in older adults. Although pharmacotherapy is the most common form of treatment, recent evidence shows cognitive-behavioural therapy to be superior in the short- and long-term management of insomnia. Low-threshold intervention programmes may reduce both the individual and societal burden of insomnia, coexisting with or without other mental or physical disorders. PMID- 17401033 TI - Therapeutic effect of follow-up assessments on antidepressant and placebo response rates in antidepressant efficacy trials: meta-analysis. AB - BACKGROUND: It remains unclear how much various factors contribute to the placebo response. AIMS: To estimate the therapeutic impact of follow-up assessments on placebo response in antidepressant trials. METHOD: Double-blind, placebo controlled antidepressant trials that reported weekly changes in Hamilton Rating Scale for Depression (HRSD) scores over 6 weeks were selected. Included studies (n=41) were divided into those that conducted four, five or six follow-up assessments. Reductions in HRSD scores as a function of the different follow-up schedules were compared. RESULTS: An extra follow-up visit at week 3 was associated with a 0.86 further reduction in HRSD score; an extra visit at week 5 was associated with a 0.67 further reduction. These effects represented approximately 34-44% of the placebo response that occurred over these time frames. Two additional visits were associated with twice the reduction in HRSD score than one, suggesting that the therapeutic impact of assessment visits is cumulative and proportional. A comparable therapeutic effect was also found in participants receiving active medication. CONCLUSIONS: Follow-up assessments in antidepressant treatment trials incur a significant therapeutic effect for participants on placebo, and this represents about 40% of the placebo response. PMID- 17401034 TI - Depression and socio-economic risk factors: 7-year longitudinal population study. AB - BACKGROUND: Low socio-economic status is associated with a higher prevalence of depression, but it is not yet known whether change in socio-economic status leads to a change in rates of depression. AIMS: To assess whether longitudinal change in socio-economic factors affects change of depression level. METHOD: In a prospective cohort study using the annual Belgian Household Panel Survey (1992 1999), depression was assessed using the Global Depression Scale. Socio-economic factors were assessed with regard to material standard of living, education, employment status and social relationships. RESULTS: A lowering in material standard of living between annual waves was associated with increases in depressive symptoms and caseness of major depression. Life circumstances also influenced depression. Ceasing to cohabit with a partner increased depressive symptoms and caseness, and improvement in circumstances reduced them; the negative effects were stronger than the positive ones. CONCLUSIONS: The study showed a clear relationship between worsening socio-economic circumstances and depression. PMID- 17401035 TI - Population level of unmet need for mental healthcare in Europe. AB - BACKGROUND: The high prevalence of mental disorders has fuelled controversy about the need for mental health services. AIMS: To estimate unmet need for mental healthcare at the population level in Europe. METHOD: As part of the European Study of Epidemiology of Mental Disorders (ESEMeD) project, a cross-sectional survey was conducted of representative samples of the adult general population of Belgium, France, Germany, Italy, The Netherlands and Spain (n=8796). Mental disorders were assessed with the Composite International Diagnostic Interview 3.0. Individuals with a 12-month mental disorder that was disabling or that had led to use of services in the previous 12 months were considered in need of care. RESULTS: About six per cent of the sample was defined as being in need of mental healthcare. Nearly half (48%) of these participants reported no formal healthcare use. In contrast, only 8% of the people with diabetes had reported no use of services for their physical condition. In total, 3.1% of the adult population had an unmet need for mental healthcare. About 13% of visits to formal health services were made by individuals without any mental morbidity. CONCLUSIONS: There is a high unmet need for mental care in Europe, which may not be eliminated simply by reallocating existing healthcare resources. PMID- 17401036 TI - Social problem-solving plus psychoeducation for adults with personality disorder: pragmatic randomised controlled trial. AB - BACKGROUND: Social problem-solving therapy may be relevant in the treatment of personality disorder, although assessments of its effectiveness are uncommon. AIMS: To determine the effectiveness of a problem-solving intervention for adults with personality disorder in the community under conditions resembling routine clinical practice. METHOD: Participants were randomly allocated to brief psychoeducation plus 16 problem-solving group sessions (n=87) or to waiting-list control (n=89). Primary outcome was comparison of scores on the Social Problem Solving Inventory and the Social Functioning Questionnaire between intervention and control arms at the conclusion of treatment, on average at 24 weeks after randomisation. RESULTS: In intention-to-treat analysis, those allocated to intervention showed significantly better problem-solving skills (P<0.001), higher overall social functioning (P=0.031) and lower anger expression (P=0.039) compared with controls. No significant differences were found on use of services during the intervention period. CONCLUSIONS: Problem-solving plus psychoeducation has potential as a preliminary intervention for adults with personality disorder. PMID- 17401037 TI - Fair assessment of the merits of psychiatric research. AB - BACKGROUND: Use of bibliometric assessments of research quality is growing worldwide. So far, a narrow range of metrics have been applied across the whole of biomedical research. Without specific sets of metrics, appropriate to each sub field of research, biased assessments of research excellence are possible. AIMS: To discuss the measures used to evaluate the merits of psychiatric biomedical research, and to propose a new approach using a multidimensional selection of metrics appropriate to each particular field of medical research. METHOD: Three steps: (a) a definition of scientific 'domains', (b) translating these into 'filters' to identify publications from bibliometric databases, leading to (c) the creation of standardised measures of merit. RESULTS: We propose using: (a) established metrics such as impact factors and citation indices, (b) new derived measures such as the 'worldscale' score, and (c) new indicators based on journal peer esteem, impact on clinical practice, medical education and health policy. CONCLUSIONS: No single index or metric can be used as a fair rating to compare nations, universities, research groups, or individual investigators across biomedical science. Rather, we propose using a multidimensional profile composed of a carefully selected array of such metrics. PMID- 17401038 TI - Psychiatric disorder among British children looked after by local authorities: comparison with children living in private households. AB - BACKGROUND: Children looked after by local authorities are at higher risk of poor psychosocial outcomes than children living in private households, but nationally representative and random samples of the two groups of children have not previously been compared. AIMS: To find explanations for the increased prevalence of psychiatric disorder in children looked after by local authorities. METHOD: We examined socio-demographic characteristics and psychopathology by type of placement among children looked after in Britain by local authorities (n=1453), and compared these children with deprived and non-deprived children living in private households (n=10 428). RESULTS: Children looked after by local authorities had higher levels of psychopathology, educational difficulties and neurodevelopmental disorders, and 'looked after' status was independently associated with nearly all types of psychiatric disorder after adjusting for these educational and physical factors. The prevalence of psychiatric disorder was particularly high among those living in residential care and with many recent changes of placement. CONCLUSIONS: Our findings indicate a need for greater support of this vulnerable group of children. PMID- 17401039 TI - Impact of psychiatric disorders on health-related quality of life: general population survey. AB - BACKGROUND: Measurement of health-related quality of life (HRQoL) with generic preference-based instruments enables comparisons of severity across different conditions and treatments. This is necessary for rational public health policy. AIMS: To measure HRQoL decrement and loss of quality-adjusted life-years (QALYs) associated with pure and comorbid forms of depressive and anxiety disorders and alcohol dependence. METHOD: A general population survey was conducted of Finns aged 30 years and over. Psychiatric disorders were diagnosed with the Composite International Diagnostic Interview and HRQoL was measured with the 15D and EQ-5D questionnaires. RESULTS: Dysthymia, generalised anxiety disorder and social phobia were associated with the largest loss of HRQoL on the individual level before and after adjusting for somatic and psychiatric comorbidity. On the population level, depressive disorders accounted for 55%, anxiety disorders 30%, and alcohol dependence for 15% of QALY loss identified in this study. CONCLUSIONS: Chronic anxiety disorders and dysthymia are associated with poorer HRQoL than previously thought. PMID- 17401040 TI - Burden on caregivers of people with schizophrenia: comparison between Germany and Britain. AB - BACKGROUND: Burden on the relatives of patients with schizophrenia may be influenced not only by patient and caregiver characteristics, but also by differences in mental health service provision. AIMS: To analyse whether family burden is affected by national differences in the provision of mental health services. METHOD: Patients with schizophrenia and their key relatives were examined in Germany (n=333) and Britain (n=170). Differences in family burden in both countries were analysed with regression models controlling for patient and caregiver characteristics. RESULTS: Family burden was associated with patients'symptoms, male gender, unemployment and marital status, as well as caregivers'coping abilities, patient contact and being a patient's parent. However, even when these attributes were controlled for, British caregivers reported more burden than German caregivers. CONCLUSIONS: National differences in family burden may be related to different healthcare systems in Germany and Britain. Support for patients with schizophrenia may be shifted from the professional to the informal healthcare sector more in Britain than in Germany. PMID- 17401041 TI - Association between trauma exposure and delusional experiences in a large community-based sample. AB - BACKGROUND: Surveys have found that otherwise well individuals report delusional experiences. Previous studies have shown an association between psychotic symptoms and exposure to trauma. AIMS: To explore the association between trauma and delusional experiences in a community sample. METHOD: Respondents (n=10 641) were assessed for delusional experiences, exposure to various types of traumatic experiences and the presence of post-traumatic stress disorder (PTSD). The endorsement of delusional items was examined in people exposed to traumatic events who did or did not have PTSD. RESULTS: Exposure to any traumatic event but without the development of PTSD was associated with increased endorsement of delusional experiences (relative risk 2.68, 95% CI 2.18-3.30) and there was a significant dose-response relationship between the number of types of traumatic events and endorsement of such experiences (chi(2) =26.74, d.f.=2, P<0.001). A diagnosis of PTSD further increased endorsement of delusional experiences (RR=9.24, 95% CI 6.95-12.27). The association between PTSD and delusional experiences remained significant after adjusting for factors associated with psychotic symptoms. CONCLUSIONS: Further investigation into the pathways between trauma, delusions and psychosis may provide insights into shared aetiological mechanisms underpinning these conditions. PMID- 17401042 TI - Effect of comorbid anxiety on treatment response and relapse risk in late-life depression: controlled study. AB - BACKGROUND: Comorbid anxiety is common in depressive disorders in both middle and late life, and it affects response to antidepressant treatment. AIMS: To examine whether anxiety symptoms predict acute and maintenance (2 years) treatment response in late-life depression. METHOD: Data were drawn from a randomised double-blind study of pharmacotherapy and interpersonal psychotherapy for patients age 70 years and over with major depression. Anxiety symptoms were measured using the Brief Symptom Inventory. Survival analysis tested the effect of pre-treatment anxiety on response and recurrence. RESULTS: Patients with greater pretreatment anxiety took longer to respond to treatment and had higher rates of recurrence. Actuarial recurrence rates were 29% (pharmacotherapy, lower anxiety), 58% (pharmacotherapy, higher anxiety), 54% (placebo, lower anxiety) and 81% (placebo, higher anxiety). CONCLUSIONS: Improved identification and management of anxiety in late-life depression are needed to achieve response and stabilise recovery. PMID- 17401043 TI - Enduring consequences of terrorism: 7-month follow-up survey of reactions to the bombings in London on 7 July 2005. AB - BACKGROUND: Terrorist attacks can have psychological effects on the general public. AIMS: To assess the medium-term effects of the July 2005 London bombings on the general population in London and to identify risk factors for persistent effects. METHOD: We telephoned 1010 Londoners 11-13 days after the bombings to assess stress levels, perceived threat and travel intentions. Seven months later, 574 respondents were contacted again and asked similar questions, and questions concerning altered perceptions of self and the world. RESULTS: ;Substantial stress' (11%), perceived threat to self (43%) and reductions in travel because of the bombings (19%) persisted at a reduced level; other perceived threats remained unchanged. A more negative world view was common. Other than degree of exposure to the bombings, there were no consistent predictors of which people with short term reactions would develop persistent reactions. CONCLUSIONS: A longer-term impact of terrorism on the perceptions and behaviour of Londoners was documented. PMID- 17401044 TI - Hypothalamic-pituitary-adrenal axis response in borderline personality disorder without post-traumatic features. AB - Hypothalamic-pituitary-adrenal (HPA) axis sensitivity was investigated in 32 non medicated patients with borderline personality disorder without comorbid post traumatic syndromes and in 18 normal individuals using a modified dexamethasone suppression test (0.25 mg). Enhanced cortisol suppression was found in the patients v. controls (P<0.05) and the percentage of participant's with non suppression was smaller in the patient (34%) than in the control group (89%) (P<0.01). Baseline cortisol levels in the patients were also lower than in the controls (P<0.05). The 0.25 mg dexamethasone suppression test reveals increased feedback inhibition of the HPA in borderline personality disorder. PMID- 17401045 TI - Lithium and risk for Alzheimer's disease in elderly patients with bipolar disorder. AB - Bipolar disorder is associated with increased risk for dementia. We compared the prevalence of Alzheimer's disease between 66 elderly euthymic patients with bipolar disorder who were on chronic lithium therapy and 48 similar patients without recent lithium therapy. The prevalence of dementia in the whole sample was 19% v. 7% in an age-comparable population. Alzheimer's disease was diagnosed in 3 patients (5%) on lithium and in 16 patients (33%) who were not on lithium (P<0.001). Our case-control data suggest that lithium treatment reduced the prevalence of Alzheimer's disease in patients with bipolar disorder to levels in the general elderly population. This is in accordance with reports that lithium inhibits crucial processes in the pathogenesis of Alzheimer's disease. PMID- 17401047 TI - Symptom dimensions and the Kraepelinian dichotomy. PMID- 17401049 TI - Major depression' in Ethiopia: validity is the problem. PMID- 17401050 TI - Prion disease in Sri Lanka. PMID- 17401052 TI - Suicide risk and perinatal circumstances. PMID- 17401054 TI - Agoraphobia in an evolutionary context. PMID- 17401055 TI - Outcome and impact of mental disorders in primary care at 5 years. AB - OBJECTIVE: To assess 5-year mental disorder recognition rates and determine the natural history of mental disorders in primary care. METHODS: A prospective cohort of adults presenting to a primary care walk-in clinic with a physical symptom were evaluated at baseline (n = 500) and at 5 years (n = 387) for mental disorders with the Primary Care Evaluation of Mental Disorders (PRIME-MD). Additional measures included functional status (Medical Outcomes Study SF-6; MOS SF6), Patient Health Questionnaire-15, Satisfaction (Rand-9), unmet expectations, and symptom outcome. Patients self-reported whether their disorder was diagnosed or treated at the 5-year follow-up. RESULTS: At baseline, 29% of patients had a mental disorder (major depression: 8.4%, minor depression 10.4%, Panic disorder 1.4%, generalized anxiety disorder 2%, anxiety not otherwise specified (NOS) 11.4%); of these patients, 26% had more than one mental disorder. Over 5 years, 33% were recognized. Threshold disorders were more likely to be recognized (major depression 56%, panic 100%, generalized anxiety disorder 88%) than subthreshold disorders (minor depression 20%, anxiety NOS 25%). Correlates of recognition included having a threshold or multiple disorders; recognition was associated with greater likelihood of persistence. Most patients with subthreshold disorders at baseline had no disorder at 5 years and few progressed to threshold disorders (minor to major depression 12%, anxiety NOS to generalized anxiety or panic 8%). CONCLUSIONS: Mental disorders are common and their recognition and treatment remain low. Subthreshold disorders have a better prognosis. Patients with threshold or multiple disorders, worse functioning or persistence of their disorder were more likely to be diagnosed. PMID- 17401056 TI - Physical and mental comorbidity, disability, and suicidal behavior associated with posttraumatic stress disorder in a large community sample. AB - OBJECTIVE: To assess if posttraumatic stress disorder (PTSD), recognized as a common mental disorder in the general population and veteran samples, has a unique impact on comorbidity, disability, and suicidal behavior (after adjusting for other mental disorders, especially depression). METHODS: Data came from the Canadian Community Health Survey Cycle 1.2 (n = 36,984; age > or = 15 years; response rate 77%). All respondents were asked if they had been given a diagnosis of PTSD by a healthcare professional. A select number of mental disorders were assessed by the Composite International Diagnostic Interview. Chronic physical health conditions, measures of quality of life, disability, and suicidal behavior were also assessed. RESULTS: The prevalence of PTSD as diagnosed by health professionals was 1.0% (95% CI = 0.90-1.15). After adjusting for sociodemographic factors and other mental disorders, PTSD remained significantly associated with several physical health problems including cardiovascular diseases, respiratory diseases, chronic pain conditions, gastrointestinal illnesses, and cancer. After adjusting for sociodemographic factors, mental disorders, and severity of physical disorders, PTSD was associated with suicide attempts, poor quality of life, and short- and long-term disability. CONCLUSIONS: PTSD was uniquely associated with several physical disorders, disability, and suicidal behavior. Increased early recognition and treatment of PTSD are warranted. PMID- 17401057 TI - Depressive symptoms, omega-6:omega-3 fatty acids, and inflammation in older adults. AB - OBJECTIVE: To address how interactions between polyunsaturated fatty acid (PUFA) levels and depressive symptoms were related to proinflammatory cytokine synthesis. Depression and stress promote proinflammatory cytokine production. Dietary intakes of omega-3 (n-3) and omega-6 (n-6) PUFAs also influence inflammation; high n-6:n-3 ratios enhance proinflammatory cytokine production, although n-3 has anti-inflammatory properties. METHODS: Blood samples from 43 older adults (mean age = 66.67 years, SD = 10.09) provided data on PUFAs and tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-6 soluble receptor (sIL-6r). Depressive symptoms were assessed by the Center for Epidemiological Studies Depression Scale. RESULTS: Depressive symptoms and n-6:n-3 ratios worked together to enhance proinflammatory cytokines beyond the contribution provided by either variable alone, with substantial variance explained by their interaction: 13% for IL-6 and 31% for TNF-alpha, whereas full models accounted for 18% and 40%, respectively. Although predicted cytokine levels were consistent across n 6:n-3 ratios with low depressive symptoms, higher n-6:n-3 ratios were associated with progressively elevated TNF-alpha and IL-6 levels as depressive symptoms increased. Higher levels of sIL-6r were associated with higher n-6:n-3 ratios. Six individuals who met the criteria for major depressive disorder had higher n 6:n-3 ratios and TNF-alpha, IL-6, and sIL-6r levels than those who did not meet the criteria; excluding these six individuals reduced the variance explained by the depressive symptoms and n-6:n-3 ratio interaction. CONCLUSIONS: Diets with high n-6:n-3 PUFA ratios may enhance the risk for both depression and inflammatory diseases. PMID- 17401058 TI - Immune function declines with unemployment and recovers after stressor termination. AB - OBJECTIVE: To examine the effect of unemployment on natural killer cell cytotoxicity (NKCC) and, in a subsample of persons who become re-employed, to determine if, after termination of the stressor, immune values recover to levels similar to matched controls. METHODS: One hundred unemployed and 100 matched employed healthy men and women, aged 29 to 45 years, were followed for 4 months with monthly blood samples taken to measure NKCC, the ability of NK cells to kill target cells. Twenty-five participants obtained employment before the end of the study, leaving 75 unemployed (and 75 employed) participants in the main sample. For unemployed participants who obtained employment before the end of the study, subsample analyses compared NKCC levels before and after obtaining a new job. RESULTS: The persistently unemployed sample had significantly lower NKCC levels for all three effector:target ratios (100:1, p = .0004; 50:1, p = .002; and 25:1, p = .02) when compared with the matched employed sample. There were no significant gender effects. In the subsample analyses, NKCC was significantly higher after the participants became employed, compared with their unemployed period, with substantial "recovery" of immune function (44%-72%) compared with values from the steadily employed group. CONCLUSIONS: Chronic stress is associated with persistent NKCC impairment. When the chronic stressor is terminated, however, the immune cell functional capacity quickly begins to recover. We believe this is the first study in humans to document immune function recovery after the definable end of a chronic stressor. PMID- 17401059 TI - Personality and medical illness burden among older adults in primary care. AB - OBJECTIVE: To examine the association between Five Factor Model personality traits (Neuroticism, Extraversion, Openness to experience, Agreeableness, Conscientiousness) and physician-quantified aggregate morbidity in a sample of older adults in primary care. METHODS: A total of 449 primary care patients, ranging in age from 65 to 97 years (75 +/- 6.9 (mean +/- standard deviation)), completed the Neo-Five Factor Inventory (NEO-FFI) and extensive interviews. A physician-investigator completed the Cumulative Illness Rating Scale (CIRS), a well-validated measure of aggregate morbidity based on a review of medical records. RESULTS: Bivariate analyses demonstrated that all five domains of the NEO-FFI were associated with CIRS scores. Multivariate regression controlling for age, gender, education, depression, smoking, hypertension, total cholesterol, alcohol or substance misuse, and other personality traits showed that greater Conscientiousness was independently associated with lower CIRS scores (beta = 0.10, t(435) = -1.96, p = .05). Other independent predictors of less morbidity were younger age, absence of hypertension, and lower levels of depression. CONCLUSION: Our results point toward the necessity of considering Conscientiousness and other personality traits in studies of risk factors for aggregate morbidity. More detailed characterization of at-risk populations will increase the likelihood of constructing informed and effective prevention, intervention, and policy initiatives. PMID- 17401060 TI - The greying of cardiology: implications for management. AB - "The development of new therapies and devices should reflect the dominant population that will ultimately use them--in the case of cardiovascular therapies and devices, this is exemplified by elderly patients" PMID- 17401061 TI - Non-invasive tests in coronary artery disease: are we facing a fork in the road? PMID- 17401062 TI - Value of rapid-access chest pain clinics. AB - Rapid-access chest pain clinics have contributed enormously to improvements under the NSF coronary heart disease banner. PMID- 17401063 TI - A post-infarction myocardium sinus. PMID- 17401064 TI - Closure of a large patent ductus arteriosus in adults: first do no harm. PMID- 17401065 TI - Role of non-invasive imaging in the management of coronary artery disease: an assessment of likely change over the next 10 years. A report from the British Cardiovascular Society Working Group. AB - Coronary angiography has been the gold standard for determining the severity, extent and prognosis of coronary atheromatous disease for the past 15-20 years. However, established non-invasive testing (such as myocardial perfusion scintigraphy and stress echocardiography) and newer imaging modalities (multi detector x ray computed tomography and cardiovascular magnetic resonance) now need to be considered increasingly as a challenge to coronary angiography in contemporary practice. An important consideration is the degree to which appropriate use of such techniques impacts on the need for coronary angiography over the next 10-15 years. This review aims to determine the role of the various investigation techniques in the management of coronary artery disease and their resource implications, and should help determine future service provision, accepting that we are in a period of significant technological change. PMID- 17401066 TI - Multidetector computed tomography image of apical left ventricular descending aorta conduit. PMID- 17401067 TI - Giant left atrium and a right sided heart. PMID- 17401068 TI - Dynamic collateral coronary circulation: angiographic evidence. PMID- 17401069 TI - 64 multidetector CT in patent foramen ovale. PMID- 17401070 TI - Rescue saphenous vein graft, 19 years on. PMID- 17401071 TI - Dynamic left main coronary artery stenosis caused by an aortic aneurysm after aortic valve replacement. PMID- 17401072 TI - Disease monitoring of patients with chronic heart failure. PMID- 17401073 TI - Infective endocarditis: a comparison of international guidelines. PMID- 17401074 TI - The heart in Anderson-Fabry disease and other lysosomal storage disorders. PMID- 17401075 TI - Split coronary artery seen with computed tomography and magnetic resonance imaging. PMID- 17401076 TI - Nuclear medicine scientists: findings and recommendations based on a 2006 survey. PMID- 17401078 TI - Cardiovascular molecular imaging. PMID- 17401079 TI - ABMS requires lifelong learning. PMID- 17401080 TI - Moving professionally. PMID- 17401081 TI - Access to medical imaging act of 2007 introduced. PMID- 17401082 TI - PET is alive and well. PMID- 17401083 TI - Toward patient-friendly cell-level dosimetry. PMID- 17401084 TI - Somatostatin receptor imaging in patients with neuroendocrine tumors: not only SPECT? PMID- 17401085 TI - Diagnostic accuracy of 18F-FDG PET in restaging patients with medullary thyroid carcinoma and elevated calcitonin levels. AB - Medullary thyroid carcinoma (MTC) is a rare endocrine tumor arising from the C cells of the thyroid gland. Calcitonin is the principal serum tumor marker. A rising calcitonin level after total thyroidectomy for localized disease generally indicates residual, recurrent, or metastatic disease. The role of (18)F-FDG PET in MTC remains somewhat unclear. We reviewed our own experience with (18)F-FDG PET in postthyroidectomy MTC patients with elevated calcitonin. METHODS: From our database, we identified patients with suspected residual, recurrent, or metastatic MTC and elevated calcitonin who had been referred for (18)F-FDG PET between January 2000 and October 2005. (18)F-FDG PET findings were classified as positive or negative on the basis of visual interpretation of the scan. Standardized uptake values (SUVs) were also calculated. The (18)F-FDG PET findings were verified by histopathologic examination, when available, or other imaging studies and clinical follow-up. Any negative (18)F-FDG PET result was considered false-negative. RESULTS: Twenty-eight patients underwent a total of 38 (18)F-FDG PET studies. Calcitonin levels ranged from 106 to 541,000 pg/mL (median, 7,260 pg/mL). There were 23 true-positive, 1 false-positive, and 14 false-negative (18)F-FDG PET scans, yielding an overall sensitivity of 62%. There was no true-positive finding when calcitonin levels were below 509 pg/mL (n = 5). Using an arbitrary cutoff of 1,000 pg/mL, we found that the sensitivity in scans with calcitonin levels greater than 1,000 pg/mL increased to 78% (21/27; 95% confidence interval, 58%-91%). The mean SUV of all lesions with (18)F-FDG uptake was 5.3 +/- 3.2 (range, 2.0-15.9). Among the 14 patients with false-negative (18)F-FDG PET findings, 8 had concurrent anatomic imaging studies and only 2 of these had positive findings. CONCLUSION: (18)F-FDG PET can detect residual, recurrent, or metastatic MTC with a reasonable sensitivity of 78% when the calcitonin level is above 1,000 pg/mL but appears of limited use if the calcitonin level is below 500 pg/mL. PMID- 17401086 TI - 68Ga-DOTA-Tyr3-octreotide PET in neuroendocrine tumors: comparison with somatostatin receptor scintigraphy and CT. AB - The aim of this study was to evaluate the diagnostic value of a new somatostatin analog, (68)Ga-labeled 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid-d-Phe(1)-Tyr(3)-octreotide ((68)Ga-DOTA-TOC), for PET in patients with known or suspected neuroendocrine tumors. PET was compared with conventional scintigraphy and dedicated CT. METHODS: Eighty-four patients (48 men, 36 women; age range, 28-79 y; mean age +/- SD, 58.2 +/- 12.2 y) were prospectively studied. For analysis, patients were divided into 3 groups: detection of unknown primary tumor in the presence of clinical or biochemical suspicion of neuroendocrine malignancy (n = 13 patients), initial tumor staging (n = 36 patients), and follow up after therapy (n = 35 patients). Each patient received 100-150 MBq (68)Ga-DOTA TOC. Imaging results of PET were compared with (99m)Tc-labeled hydrazinonicotinyl Tyr(3)-octreotide ((99m)Tc-HYNIC-TOC) and (111)In-DOTA-TOC. CT was also performed on every patient using a multidetector scanner. Each imaging modality was interpreted separately by observers who were unaware of imaging findings before comparison with PET. The gold standard for defining true-positive (TP), true negative (TN), false-positive (FP), and false-negative (FN) results was based on all available histologic, imaging, and follow-up findings. RESULTS: PET was TP in 69 patients, TN in 12 patients, FP in 1 patient, and FN in 2 patients, indicating a sensitivity of 97%, a specificity of 92%, and an accuracy of 96%. The FP finding was caused by enhanced tracer accumulation in the pancreatic head, and the FN results were obtained in patients with a tumor of the gastrointestinal tract displaying liver metastases. (68)Ga-DOTA-TOC showed higher diagnostic efficacy compared with SPECT (TP in 37 patients, TN in 12 patients, FP in 1 patient, and FN in 34 patients) and diagnostic CT (TP in 41 patients, TN in 12 patients, FP in 5 patients, and FN in 26 patients). This difference was of statistical significance (P < 0.001). However, the combined use of PET and CT showed the highest overall accuracy. CONCLUSION: (68)Ga-DOTA-TOC PET shows a significantly higher detection rate compared with conventional somatostatin receptor scintigraphy and diagnostic CT with clinical impact in a considerable number of patients. PMID- 17401087 TI - Prognostic value of O-(2-18F-fluoroethyl)-L-tyrosine PET and MRI in low-grade glioma. AB - In glioma of World Health Organization (WHO) grade II (low-grade glioma), the natural course of a particular patient is not predictable and the treatment strategy is controversial. We determined prognostic factors in adult patients with untreated, nonenhancing, supratentorial low-grade glioma with special regard to PET using the amino acid O-(2-(18)F-fluoroethyl)-L-tyrosine ((18)F-FET) and MRI. METHODS: In a prospective study, baseline (18)F-FET PET and MRI analyses were performed on 33 consecutive patients with histologically confirmed low-grade glioma. None of the patients had radiation or chemotherapy. Clinical, histologic, therapeutic (initial cytoreduction vs. biopsy), (18)F-FET uptake, and MRI morphologic parameters were analyzed for their prognostic significance. Statistical endpoints were clinical or radiologic tumor progression, malignant transformation to glioma of WHO grade III or IV (high-grade glioma), and death. RESULTS: Baseline (18)F-FET uptake and a diffuse versus circumscribed tumor pattern on MRI were highly significant predictors of prognosis (P < 0.01). By the combination of these prognostically significant variables, 3 major prognostic subgroups of low-grade glioma patients could be identified. The first of these subgroups was patients with circumscribed low-grade glioma on MRI without (18)F FET uptake (n = 11 patients, progression in 18%, no malignant transformation and no death). The second subgroup was patients with circumscribed low-grade glioma with (18)F-FET uptake (n = 13 patients, progression in 46%, malignant transformation to a high-grade glioma in 15%, and death in 8%). The third subgroup was patients with diffuse low-grade glioma with (18)F-FET uptake (n = 9 patients, progression in 100%, malignant transformation to a high-grade glioma in 78%, and death in 56%). CONCLUSION: We conclude that baseline amino acid uptake on (18)F-FET PET and a diffuse versus circumscribed tumor pattern on MRI are strong predictors for the outcome of patients with low-grade glioma. PMID- 17401088 TI - Improving specificity of breast MRI using prone PET and fused MRI and PET 3D volume datasets. AB - MRI is a sensitive method for detecting invasive breast cancer, but it lacks specificity. To examine the effect of combining PET with MRI on breast lesion characterization, a prototype positioning device was fabricated to allow PET scans to be acquired in the same position as MRI scans--that is, prone. METHODS: To test the hypothesis that fusion of (18)F-FDG PET and MRI scans improves detection of breast cancer, 23 patients with suspected recurrent or new breast cancer underwent a routine whole-body PET scan, a prone PET scan of the chest, and a routine breast MRI scan. The attenuation-corrected prone PET and MRI datasets were registered twice by different operators. The fusion results were judged for quality by visual inspection and statistical analysis. A joint reading of the MRI and PET scans side by side and integrated images was performed by a nuclear medicine physician and a radiologist. Sensitivity and specificity of MRI and combined MRI and PET scans were calculated on the basis of pathology reports or at least 1 y of clinical and radiologic follow-up. RESULTS: All fusions were verified to be well matched using specific anatomic criteria. A total of 45 lesions was assessed. Lesion size range was 0.6 to 10.0 cm. Of the 44 breasts examined, 29 were suspicious for cancer, of which 15 were found to be positive on surgical excision. In lesion-by-lesion analysis, sensitivity and specificity of MRI alone were 92% and 52%, respectively; after MRI and PET fusion, they were 63% and 95%, respectively. The positive predictive value and the negative predictive value for MRI alone were 69% and 85%, respectively; after MRI and PET fusion, they were 94% and 69%, respectively. CONCLUSION: Acquisition of prone PET scans using the new positioning device permitted acquisition of prone scans suitable for fusion with breast MRI scans. Fused PET and MRI scans increased the specificity of MRI but decreased the sensitivity in this small group of patients. Additional data are needed to confirm the statistical significance of these preliminary findings. PMID- 17401089 TI - Assessment of 11C-PE2I binding to the neuronal dopamine transporter in humans with the high-spatial-resolution PET scanner HRRT. AB - The high-resolution research tomograph (HRRT), dedicated to brain imaging, may offer new perspectives for identifying small brain nuclei that remain neglected by the spatial resolution of conventional scanners. However, the use of HRRT for neuroimaging applications still needs to be fully assessed. The present study aimed at evaluating the HRRT for measurement of the dopamine transporter (DAT) binding to validate its quantification and explore the gain induced by the increased spatial resolution in comparison with conventional PET scanners. METHODS: Fifteen and 11 healthy subjects were examined using the selective DAT radioligand (11)C-PE2I with HRRT and HR+ scanners, respectively. Quantification of the DAT binding was assessed by the calculation of binding potential (BP) values using the simplified reference tissue model in anatomic regions of interest (ROIs) defined on the dorsal striatum and in a standardized ROI defined on the midbrain. RESULTS: Quantification of (11)C-PE2I binding to the DAT measured in the midbrain and striatum with both scanners at the same spatial resolution (smoothed HRRT images) exhibited similar BP values and intersubject variability, thus validating the quantification of DAT binding on the HRRT. For age-paired comparison, BP values of subjects examined with HRRT were significantly higher than those of the subjects examined with HR+. The increase ranged from 29% in the caudate and 35% in the putamen to 92% in the midbrain. The decline in DAT binding with age in the striatum was in good agreement between both scanners and literature, whereas no significant decrease in DAT binding with age was observed in the midbrain with either HRRT or HR+. CONCLUSION: HRRT allows quantitative measurements of neurotransmission processes in small brain nuclei and allows recovering higher values as compared with coarser spatial resolution PET scanners. High-spatial-resolution PET appears promising for a more accurate detection of neurobiologic modifications and also for the exploration of subtle modifications in small and complex brain structures largely affected by the partial-volume effect. PMID- 17401090 TI - Visual assessment versus quantitative assessment of 11C-PIB PET and 18F-FDG PET for detection of Alzheimer's disease. AB - Amyloid-beta (Abeta) imaging with N-methyl-(11)C-2-(4'-methylamino-phenyl)-6 hydroxy-benzothiazole ((11)C-6-OH-BTA-1; also known as (11)C-PIB) shows a robust increase in cortical binding in Alzheimer's disease (AD). The aim of this study was to explore the clinical potential of Abeta imaging for the diagnosis of AD by comparison of the accuracy of visual reading of (11)C-PIB images with quantitative analysis and (18)F-FDG. METHODS: Fifteen AD patients (age, 71.1 +/- 11.3 y [mean +/- SD]; mini-mental state examination [MMSE], 18.9 +/- 9.3 [mean +/ SD]) and 25 healthy control (HC) subjects (age, 71.9 +/- 6.82 y; MMSE >or= 28) underwent 90-min dynamic (11)C-PIB PET and 20-min static (18)F-FDG PET. (11)C-PIB images, generated from data acquired between 40 and 70 min after injection, and (18)F-FDG images were rated separately by 2 readers as normal, possible AD, or probable AD. Quantitative analyses used the distribution volume ratio (DVR) of frontal cortex, parietotemporal cortex, posterior cingulate, and caudate nucleus for (11)C-PIB and standardized uptake value ratio (SUVR) of parietotemporal cortex and posterior cingulate for (18)F-FDG, using cerebellar cortex as the reference region. Receiver-operating-characteristic (ROC) analysis was performed to compare the accuracy of quantitative measures. To determine the effect of age on diagnostic accuracy, the median age of the AD subjects (74 y) was chosen to separate the cohort into younger (64.4 +/- 5.8 y) and older (78.6 +/- 4.1 y) groups. RESULTS: Visual agreement between readers was excellent for (11)C-PIB (kappa = 0.90) and good for (18)F-FDG (kappa = 0.56). (11)C-PIB was more accurate than (18)F-FDG both on visual reading (accuracy, 90% vs. 70%, P = 0.05) and ROC analysis (95% vs. 83%, P = 0.02). Accuracy declined more with (18)F-FDG than with (11)C-PIB in the older group. CONCLUSION: Visual analysis of (11)C-PIB images appears more accurate than visual reading of (18)F-FDG for identification of AD and has accuracy similar to quantitative analysis of a 90-min dynamic scan. The accuracy of (11)C-PIB PET is limited by cortical binding in some healthy elderly subjects, consistent with postmortem studies of cerebral Abeta. Longitudinal follow-up is required to determine if this represents detection of preclinical AD. PMID- 17401091 TI - 2-(2-[2-Dimethylaminothiazol-5-yl]ethenyl)-6- (2-[fluoro]ethoxy)benzoxazole: a novel PET agent for in vivo detection of dense amyloid plaques in Alzheimer's disease patients. AB - Extensive deposition of dense amyloid fibrils is a characteristic neuropathologic hallmark in Alzheimer's disease (AD). Noninvasive detection of these molecules is potentially useful for early and precise detection of patients with AD. This study reports a novel compound, 2-(2-[2-dimethylaminothiazol-5-yl]ethenyl)-6-(2 [fluoro]ethoxy)benzoxazole (BF-227), for in vivo detection of dense amyloid deposits using PET. METHODS: The binding affinity of BF-227 to amyloid-beta (Abeta) fibrils was calculated. The binding property of BF-227 to amyloid plaques was evaluated by neuropathologic staining of AD brain sections. Brain uptake and in vivo binding of BF-227 to Abeta deposits were also evaluated using mice. For clinical evaluation of (11)C-BF-227 as a PET probe, 11 normal (healthy) subjects and 10 patients with AD participated in this study. Dynamic PET images were obtained for 60 min after administration of (11)C-BF-227. The regional standardized uptake value (SUV) and the ratio of regional to cerebellar SUV were calculated as an index of (11)C-BF-227 retention. The regional tracer distribution in AD patients was statistically compared with that of aged normal subjects on a voxel-by-voxel basis. RESULTS: BF-227 displayed high binding affinity to synthetic Abeta1-42 fibrils (K(i) [inhibition constant], 4.3 +/- 1.5 nM). Neuropathologic staining has demonstrated preferential binding of this agent to dense amyloid deposits in AD brain. Moreover, a biodistribution study of this agent revealed excellent brain uptake and specific labeling of amyloid deposits in transgenic mice. The present clinical PET study using (11)C-BF-227 demonstrated the retention of this tracer in cerebral cortices of AD patients but not in those of normal subjects. All AD patients were clearly distinguishable from normal individuals using the temporal SUV ratio. Voxel-by-voxel analysis of PET images revealed that cortical BF-227 retention in AD patients is distributed primarily to the posterior association area of the brain and corresponded well with the preferred site for neuritic plaque depositions containing dense Abeta fibrils. CONCLUSION: These findings suggest that BF-227 is a promising PET probe for in vivo detection of dense amyloid deposits in AD patients. PMID- 17401092 TI - Noninvasive detection of programmed cell loss with 99mTc-labeled annexin A5 in heart failure. AB - Apoptosis, or programmed cell death (PCD), contributes to the decline in ventricular function in heart failure. Because apoptosis comprises a programmed cascade of events, it is potentially reversible, and timely intervention should delay the development of cardiomyopathy. (99m)Tc-Labeled annexin A5 has successfully been used for the noninvasive detection of PCD in myocardial infarction and heart transplant rejection. The present study evaluated the role of annexin A5 imaging for detection of PCD in heart failure patients. METHODS: Annexin A5 imaging was performed on 9 consecutive heart failure patients with advanced nonischemic cardiomyopathy (dilated, n = 8; hypertrophic, n = 1) and in 2 relatives having the same genetic background as the hypertrophic cardiomyopathy patient but no heart failure. RESULTS: Four of the patients with dilated cardiomyopathy and the 1 with hypertrophic cardiomyopathy and heart failure showed focal, multifocal, or global left ventricular uptake of annexin A5. No uptake was visualized in the remaining 4 patients or in the 2 controls. All cases showing annexin A5 uptake within the left ventricle experienced significant reduction in left ventricular function or functional class. In cases with no annexin A5 uptake, left ventricular function and clinical status remained stable. CONCLUSION: These data indicate the feasibility of noninvasive PCD detection with annexin imaging in heart failure patients. Annexin A5 uptake is associated with deterioration in left ventricular function, and this association may lend itself to the development of novel management strategies. PMID- 17401093 TI - Experience with a simplified, standardized 4-hour gastric-emptying protocol. AB - Gastric-emptying studies have no accepted standard methodology or reference values. A simplified standardized protocol with a large reference database has been proposed, with imaging obtained at only 0, 1, 2, and 4 h. The rationale for its 4-h length is data suggesting that delayed emptying is detected with higher sensitivity at 4 h than at 2 h. The purpose of the current investigation was to review our 2-y experience using this protocol, to determine the added value of 4 h imaging, and to determine whether the lag phase can predict delayed emptying. METHODS: After ingesting a sandwich made with (99m)Tc-sulfur colloid egg substitute, 175 patients were imaged immediately, every 10 min for 1 h, and then at 2, 3, and 4 h. Percentage retention was calculated for each interval. Sensitivities, specificities, predictive values, and receiver operating characteristic (ROC) curves were computed for the 1-, 2-, and 3-h intervals on the basis of normal 4-h values. Data were analyzed to determine the added value of the 4-h study. Thresholds were sought that optimized accuracy. ROC analysis was used to determine the predictive value of the lag phase. RESULTS: Gastric emptying was delayed in 20% of patients at 2 h and in 26% at 4 h, a 29% increase in abnormal studies (P < 0.02). Of those normal at 2 h, 13% became abnormal at 4 h. Of those with delayed emptying at 2 h, 24% normalized at 4 h. Compared with the study at 4 h, the study at 2 h had a 59% sensitivity, 94% specificity, 76% PPV, and 87% NPV. ROC analysis showed that the areas under the ROC curve (AUC) were 0.75, 0.93, and 0.97 at 1, 2, and 3 h, respectively. The threshold for optimal accuracy for 3-h data was estimated to be greater than 30% retention. The accuracy of this threshold was 91%. The lag phase AUC for predicting delayed emptying at 4 h was 0.60. CONCLUSION: This first investigation of a large referral patient population using a standardized protocol found that abnormal emptying is detected with greater sensitivity at 4 h than at 2 h and that the lag phase is not predictive of delayed emptying. PMID- 17401094 TI - 11C-DPA-713: a novel peripheral benzodiazepine receptor PET ligand for in vivo imaging of neuroinflammation. AB - The induction of neuroinflammatory processes, characterized by upregulation of the peripheral benzodiazepine receptor (PBR) expressed by microglial cells, is well correlated with neurodegenerative diseases and with acute neuronal loss. The continually increasing incidence of neurodegenerative diseases in developed countries has become a major health problem, for which the development of diagnostic and follow-up tools is required. Here we investigated a new PBR ligand suitable for PET to monitor neuroinflammatory processes as an indirect hallmark of neurodegeneration. METHODS: We compared PK11195, the reference compound for PBR binding sites, with the new ligand DPA-713 (N,N-diethyl-2-[2-(4 methoxyphenyl)-5,7-dimethylpyrazolo[1,5-a]pyrimidin-3-yl]acetamide), using a small-animal dedicated PET camera in a model of neuroinflammation in rats. Seven days after intrastriatal injection of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA), a PET scan was performed using (11)C-PK11195 or (11)C DPA-713. Immunohistochemistry for neuronal (NeuN), astrocyte (glial fibrillary acidic protein), and microglial (CD11) specific markers as well as (3)H-PK11195 autoradiographic studies were then correlated with the imaging data. RESULTS: Seven days after a unilateral injection of AMPA in the striatum, (11)C-DPA-713 exhibits a better contrast between healthy and damaged brain parenchyma than (11)C-PK11195 (2.5-fold +/- 0.14 increase vs. 1.6-fold +/- 0.05 increase, respectively). (11)C-DPA-713 and (11)C-PK11195 exhibit similar brain uptake in the ipsilateral side, whereas, in the contralateral side, (11)C-DPA-713 uptake was significantly lower than (11)C-PK11195. Modeling of the data using the simplified reference tissue model shows that the binding potential was significantly higher for (11)C-DPA-713 than for (11)C-PK11195. CONCLUSION: (11)C DPA-713 displays a higher signal-to-noise ratio than (11)C-PK11195 because of a lower level of unspecific binding that is likely related to the lower lipophilicity of (11)C-DPA-713. Although further studies in humans are required, (11)C-DPA-713 represents a suitable alternative to (11)C-PK11195 for PET of PBR as a tracer of neuroinflammatory processes induced by neuronal stress. PMID- 17401095 TI - Human antibody against C domain of tenascin-C visualizes murine atherosclerotic plaques ex vivo. AB - Targeting proteins that are overexpressed in atherosclerotic plaques may open novel diagnostic applications. The C domain of tenascin-C is absent from normal adult tissues but can be inserted during tumor progression or tissue repair into the molecule by alternative splicing. We tested the ability of the human antibody G11, specific to this antigen, to reveal murine atherosclerotic plaques ex vivo. The antibody directed against the extra domain B of fibronectin (L19) was used as a reference. METHODS: We intravenously injected (125)I-labeled G11 or L19 antibodies into apolipoprotein E-deficient (ApoE(-/-)) mice and harvested the aortae 4 or 24 h later. En face analyses of distal aortae and longitudinal sections of the aortic arch were performed to compare antibody uptake using autoradiography with plaque staining using oil red O. Plaque macrophages were detected by immunohistochemistry (anti-CD68 staining). Biodistribution of injected antibodies was investigated in aortae and blood at 4 and 24 h. RESULTS: En face analyses revealed a significant correlation between radiolabeled G11 and fat-stained areas, increasing from 4 to 24 h, with a correlation coefficient of 0.92 (P < 0.0001) and an average signal-to-noise ratio of 104:1 at 24 h. Plaque imaging using L19 showed similar results (r = 0.86; P < 0.0001; signal-to-noise ratio, 72:1 at 24 h). Uptake of radiolabeled antibodies in histologic sections colocalized with fat staining and activated macrophages in aortic plaques. Biodistribution analyses confirmed specific accumulation in aortic plaques as well as rapid blood pool clearance of the antibodies 24 h after injection. Immunofluorescence analyses revealed increased expression of tenascin and fibronectin isoforms in macrophage-rich plaques. CONCLUSION: The antibody G11, specific to the C domain of tenascin-C, visualizes murine atherosclerotic plaques ex vivo. In conjunction with the increased expression of the C domain of tenascin C in macrophage-rich plaques, the colocalization of G11 uptake with activated macrophages, and the favorable target-to-blood ratio at 24 h, this antibody may be useful for molecular imaging of advanced atherosclerotic plaques in the intact organism. PMID- 17401096 TI - Development of a dual membrane protein reporter system using sodium iodide symporter and mutant dopamine D2 receptor transgenes. AB - For noninvasive monitoring of cellular status by dual reporters, a dual membrane protein reporter system was developed and its in vivo applicability was examined. Human sodium iodide symporter (hNIS) and mutant dopamine D(2) receptor (D(2)R) transgenes were chosen considering their complementarity. METHODS: pIRES hNIS/D(2)R containing NIS and D(2)R linked with an internal ribosomal entry site (IRES) was constructed and transfected into human hepatoma SK-Hep1 and rat glioma C6 cells. The cell lines stably expressing hNIS and D(2)R (named SK-ND and C6-ND) were produced, which was confirmed by messenger RNA expression of reporter genes. The functional activities of hNIS and D(2)R were measured by (125)I uptake assay and (3)H-spiperone receptor-binding assays. A biodistribution study was performed on SK-ND tumor-bearing mice using (99m)Tc-pertechnetate and (3)H-spiperone. In vivo hNIS expression was examined using (99m)Tc-pertechnetate gamma-camera imaging and, D(2)R expression was examined using a (3)H-spiperone autoradiographic study. RESULTS: (125)I uptake of SK-ND and C6-ND cell lines showed a maximum 97-fold and 43-fold increase, respectively, which were completely inhibited by KClO(4). Specific (3)H-spiperone binding to SK-ND and C6 ND cell homogenates was observed, which were completely inhibited by (+) butaclamol. Among the dual reporter gene-expressing cell lines, the activities of both reporters were inversely correlated with each other. Competition assay of hNIS-expressing cells by D(2)R vector transfection and D(2)R-expressing cells by hNIS vector transfection showed a dose-dependent decrease of hNIS and D(2)R activities, respectively. In the biodistribution study, (99m)Tc-pertechnetate accumulated 10-fold and (3)H-spiperone accumulated 4-fold more in SK-ND tumors than that in parental SK tumors. In vivo imaging of (99m)Tc-pertechnetate persisted until 5 wk after the cell graft in SK-ND tumors. Autoradiographic study of brain tissues from these mice also revealed an accumulation of (3)H-spiperone in SK-ND tumors. CONCLUSION: We developed a dual membrane-bound positron and gamma-imaging reporter system of hNIS and D(2)R. We observed its reporting capability in vitro and in vivo and elucidated that these 2 membrane protein reporters competed with each other in their expression. Although we expect that hNIS and D(2)R transgenes can complement each other as a dual reporter system, we suggest that one needs to validate the ratio of expression of the 2 membrane protein reporter transgenes for cellular status tracking. PMID- 17401097 TI - Indication for different mechanisms of kidney uptake of radiolabeled peptides. AB - Nephrotoxicity due to renal reabsorption of radiolabeled peptides limits the tumor dose in peptide receptor radiotherapy (PRRT). Therefore, we evaluated the ability of several agents to inhibit the renal accumulation of different radiopeptides. METHODS: Male Wistar rats (4 per group) were injected intravenously with 1 MBq of (111)In-labeled octreotide (OCT), minigastrin (MG), bombesin (BOM), or exendin (EX), together with a potential inhibitor of renal uptake (lysine [Lys], poly-glutamic acid [PGA], and Gelofusine [GF], a gelatin based plasma expander) or phosphate-buffered saline as a control. Organ uptake at 20 h after injection was determined as the percentage of injected activity per gram (%IA/g). Lys, PGA, and GF were also combined to determine whether an additive effect could be obtained. The localization of the peptides in the kidneys was investigated by autoradiography using a phosphor imager. RESULTS: OCT accumulation in the kidney was inhibited by Lys and GF (40.7%-45.1%), whereas PGA was ineffective. On the other hand, renal uptake of BOM, MG, and EX was inhibited by PGA and GF (15.4%-85.4%), whereas Lys was ineffective. The combination of GF and Lys showed additive effects in inhibiting OCT uptake, whereas PGA and GF had additive effects for the inhibition of EX uptake. The amount of kidney uptake correlated with the number of charged amino acids. All radiopeptides were localized in the renal cortex, as indicated by autoradiography. CONCLUSION: Inhibition of renal accumulation of the radiopeptides tested could be achieved by either Lys or PGA but not by both at the same time, suggesting 2 different uptake mechanisms. The differences in renal accumulation of radiopeptides may be related to the number of charges of a molecule. GF is the only compound that inhibited renal accumulation of all radiopeptides tested. Additional experiments are needed to further elucidate these findings and to optimize inhibition of renal accumulation of radiopeptides to reduce the kidney dose in PRRT. PMID- 17401098 TI - Reproducibility of 18F-FDG microPET studies in mouse tumor xenografts. AB - (18)F-FDG has been used to image mouse xenograft models with small-animal PET for therapy response. However, the reproducibility of serial scans has not been determined. The purpose of this study was to determine the reproducibility of (18)F-FDG small-animal PET studies. METHODS: Mouse tumor xenografts were formed with B16F10 murine melanoma cells. A 7-min small-animal PET scan was performed 1 h after a 3.7- to 7.4-MBq (18)F-FDG injection via the tail vein. A second small animal PET scan was performed 6 h later after reinjection of (18)F-FDG. Twenty five sets of studies were performed. Mean injected dose per gram (%ID/g) values were calculated from tumor regions of interest. The coefficient of variation (COV) from studies performed on the same day was calculated to determine the reproducibility. Activity from the second scans performed after 6 h were adjusted by subtracting the estimated residual activity from the first (18)F-FDG injection. For 7 datasets, an additional scan immediately before the second injection was performed, and residual activity from this additional delayed scan was subtracted from the activity of the second injection. COVs of both subtraction methods were compared. Blood glucose values were measured at the time of injection and used to correct the %ID/g values. RESULTS: The COV for the mean %ID/g between (18)F-FDG small-animal PET scans performed on the same day 6 h apart was 15.4% +/- 12.6%. The delayed scan subtraction method did not produce any significant change in the COV. Blood glucose correction increased the COV. The injected dose, tumor size, and body weight did not appear to contribute to the variability of the scans. CONCLUSION: (18)F-FDG small-animal PET mouse xenograft studies were reproducible with moderately low variability. Therefore, serial small-animal PET studies may be performed with reasonable accuracy to measure tumor response to therapy. PMID- 17401099 TI - Uptake of 18F-Fluorocholine, 18F-FET, and 18F-FDG in C6 gliomas and correlation with 131I-SIP(L19), a marker of angiogenesis. AB - Targeting extracellular structures that are involved in angiogenic processes, such as the extra domain B of fibronectin, is a promising approach for the diagnosis of solid tumors. The aim of this study was to determine uptake of the (18)F-labeled PET tracers (18)F-fluorocholine (N,N-dimethyl-N-(18)F-fluoromethyl 2-hydroxyethylammonium), (18)F-fluoro-ethyl-l-tyrosine (FET), and (18)F-FDG in C6 gliomas of the rat and to correlate it with uptake of the anti-extra domain B antibody (131)I-SIP(L19) as a marker of neoangiogenesis. METHODS: C6 gliomas were orthotopically induced in 17 rats. Uptake of all tracers was measured using quantitative autoradiography, and uptake of (18)F-fluorocholine, (18)F-FET, and (18)F-FDG was correlated with uptake of (131)I-SIP(L19) on a pixelwise basis. RESULTS: The mean (131)I-SIP(L19), (18)F-fluorocholine, (18)F-FET, and (18)F-FDG standardized uptake values in the tumor and the contralateral normal cortex (in parentheses) were 0.31 +/- 0.22 (not detectable), 2.00 +/- 0.53 (0.49 +/- 0.07), 3.67 +/- 0.36 (1.42 +/- 0.22), and 7.23 +/- 1.22 (3.64 +/- 0.51), respectively. The (131)I-SIP(L19) uptake pattern correlated best with (18)F-fluorocholine uptake (z = 0.80, averaged z-transformed Pearson correlation coefficient) and (18)F-FET uptake (z = 0.79) and least with (18)F-FDG (z = 0.37). CONCLUSION: One day after intravenous injection, (131)I-SIP(L19) displayed a very high tumor-to cortex ratio, which may be used in the diagnostic work-up of brain tumor patients. Of the 3 investigated (18)F tracers, (18)F-fluorocholine and (18)F-FET correlated better with the pattern of (131)I-SIP(L19) uptake than did (18)F-FDG. Whether this means that (18)F-fluorocholine and (18)F-FET are better suited than (18)F-FDG to monitor antiangiogenic therapy should be investigated in future studies. PMID- 17401100 TI - Selection of radiolabeled gastrin analogs for peptide receptor-targeted radionuclide therapy. AB - The gastrin/cholecystokinin-2 (CCK-2) receptor has been identified as a possible target for peptide receptor radionuclide imaging and therapy. Several radiolabeled peptides binding to this receptor have been explored in animal models and clinical trials but either low tumor uptake or high renal retention has been found. The aim of this study was to identify a peptide with improved tumor-to-kidney pharmacodynamics when compared with current candidates. METHODS: A small peptide-chelator library of 34 compounds based on the C-terminal sequences of CCK-8 or minigastrin was constructed. The peptides were radiolabeled with (111)In with high labeling efficiency (>90%), as determined by high performance liquid chromatographic analysis. The labeled peptides were screened by assessing tumor and kidney uptake in pancreatic xenograft nude mouse models, including AR42J. An extensive biodistribution analysis was performed on the lead candidate from the library. RESULTS: Minigastrin analogs containing a pentaglutamate sequence showed the highest tumor uptake but very high renal retention. CCK analogs showed the lowest tumor and renal uptake. Deletion of the pentaglutamate sequence in the gastrin analogs lowered the tumor uptake by a factor of 3 but decreased the kidney uptake by a factor of 20. Insertion of histidine residues in the sequence reduced kidney uptake by a further factor of almost 2-fold. In AR42J tumor-bearing mice, the peptide with the sequence DOTA HHEAYGWMDF-NH(2) (DOTA is tetraazacyclododecane tetraacetic acid) showed the highest tumor-to-kidney ratio of all peptides studied, with saturable uptake in target organs and low uptake by nontarget tissues other than the kidney. CONCLUSION: This peptide is a worthwhile candidate for clinical studies to determine whether it is suitable for use in peptide receptor-targeted radionuclide therapy. PMID- 17401101 TI - Precise localization of sentinel lymph nodes and estimation of their depth using a prototype intraoperative mini gamma-camera in patients with breast cancer. AB - The aim of this study was to evaluate the ability of a prototype intraoperative mini gamma-camera, the CarolIReS, with a 50 x 50 mm field of view, to precisely localize sentinel lymph nodes (SLNs) and to determine their depth in a series of patients with infiltrative breast cancer requiring SLN excision. METHODS: With the use of phantoms, the broadness of the signal of an acquired projection was shown to linearly depend on its distance from the collimator. A preclinical ex vivo study of 25 excised SLNs demonstrated that SLN size did not influence depth estimation. The minimum activity threshold for successful use of the proposed method was determined. After a preoperative radioisotope injection and lymphoscintigraphy, the SLN was localized in a series of 11 patients using both the mini gamma-camera and a gamma-probe. During surgery, a ruler was used to measure the depth of all SLNs before their excision. RESULTS: Using the measured linear dependence of image broadness, we found that the expected SLN anatomic depth was compatible with its measured depth during surgery. CONCLUSION: This study showed that the mini gamma-camera efficiently estimated the location of SLNs in 3 dimensions. PMID- 17401102 TI - Correction of heart motion due to respiration in clinical myocardial perfusion SPECT scans using respiratory gating. AB - Several studies have described nonuniform blurring of myocardial perfusion imaging (MPI) due to respiration. This article describes a technique for correcting the respiration effect and assesses its effectiveness in clinical studies. METHODS: Simulated phantoms, physical phantoms, and patient scans were used in this study. A heart phantom, which oscillated back and forth, was used to simulate respiration. The motion was measured on a gamma-camera supporting list mode functionality synchronized with an external respiratory strap or resistor sensor. Eight clinical scans were performed using a 1-d (99m)Tc-sestamibi protocol while recording the respiratory signal. The list-mode capability along with the strap or sensor signals was used to generate respiratory bin projection sets. A segmentation process was used to detect the shift between the respiratory bins. This shift was further projected to the acquired projection images for correction of the respiratory motion. The process was applied to the phantom and patient studies, and the rate of success of the correction was assessed using the conventional bull's eye maps. RESULTS: The algorithm provided a good correction for the phantom studies. The shift after the correction, measured by a fitted ellipsoid, was <1 mm in the axial direction. The average motion due to respiration in the clinical studies was 9.1 mm in the axial direction. The average shift between the respiratory phases was reduced to 0.5 mm after correction. The maximal change in the bull's eye map for the clinical scans after the correction was 6%, with a mean of 3.75%. The postcorrection clinical summed perfusion images were more uniform, consistent, and, for some patients, clinically significant when compared with the images before correction for respiration. CONCLUSION: Myocardial motion generated by respiration during MPI SPECT affects perfusion image quality and accuracy. Motion introduced by respiration can be corrected using the proposed method. The degree of correction depends on the patient respiratory pattern and can be of clinical significance in certain cases. PMID- 17401103 TI - Evaluation of 3D Monte Carlo-based scatter correction for 201Tl cardiac perfusion SPECT. AB - (201)Tl-Chloride ((201)Tl) is a myocardial perfusion SPECT agent with excellent biochemical properties commonly used for assessing tissue viability. However, cardiac (201)Tl SPECT images are severely degraded by photons scattered in the thorax. Accurate correction for this scatter is complicated by the nonuniform density and varied sizes of thoraxes, by the additional attenuation and scatter caused by female patients' breasts, and by the energy spectrum of (201)Tl. Monte Carlo simulation is a general and accurate method well suited to modeling this scatter. METHODS: Statistical reconstruction that includes Monte Carlo modeling of scatter was compared with statistical reconstruction algorithms not corrected for scatter. In the ADS method, corrections for attenuation, detector response, and scatter (Monte Carlo-based) were implemented simultaneously via the dual matrix ordered-subset expectation maximization algorithm with a Monte Carlo simulator as part of the forward projector. The ADS method was compared with the A method (ordered-subset expectation maximization with attenuation correction) and with the AD method (a method like the A method but with detector response modeling added). A dual-head SPECT system equipped with two (153)Gd scanning line sources was used for simultaneously acquiring transmission and emission data. Four clinically realistic phantom configurations (a large thorax and a small thorax, each with and without breasts) with a cardiac insert containing 2 cold defects were used to evaluate the proposed reconstruction algorithms. We compared the performance of the different algorithms in terms of noise properties, contrast-to-noise ratios, the contrast separability of perfusion defects, uniformity, and robustness to anatomic variations. RESULTS: The ADS method provided images with clearly better visual defect contrast than did the other methods. The contrasts achieved with the ADS method were 10%-24% higher than those achieved with the AD method and 11%-37% higher than those achieved with the A method. For a typical contrast level, the ADS method exhibited noise levels around 27% lower than the AD method and 34% lower than the A method. Compared with the other 2 algorithms, the ADS reconstructions were less sensitive to anatomic variations and had better image uniformity in the homogeneously perfused myocardium. Finally, we found that the improvements that can be achieved with Monte Carlo-based scatter correction are stronger for (201)Tl than for (99m)Tc imaging. CONCLUSION: Our results indicate that Monte Carlo-based scatter correction is suitable for (201)Tl cardiac imaging and that such correction simultaneously improves several image-quality metrics. PMID- 17401104 TI - Spatial distribution of blood vessels and CD34+ hematopoietic stem and progenitor cells within the marrow cavities of human cancellous bone. AB - Current bone marrow dosimetry methods inherently assume that the target cells of interest for the assessment of leukemia risk (stochastic effects) or marrow toxicity (deterministic effects) are uniformly localized throughout the marrow cavities of cancellous bone. Previous studies on mouse femur, however, have demonstrated a spatial gradient for the hematopoietic stem and progenitor cells, with higher concentrations near the bone surfaces. The objective of the present study was to directly measure the spatial concentration of these cells, as well as marrow vasculature structures, within images of human disease-free bone marrow. METHODS: Core-biopsy samples of normal bone marrow from the iliac crest were obtained from clinical cases at Shands Hospital at the University of Florida Department of Pathology. The specimens were sectioned and immunohistochemically stained for CD34 (red) and CD31 (brown) antigens. These 2 stains were used simultaneously to differentiate between hematopoietic stem and progenitor cells (CD34(+)/CD31(-)) and vascular endothelium (CD34(+)/CD31(+)). Distances from hematopoietic CD34(+) cells and blood vessels to the nearest bone trabecula surface were measured digitally and then binned in 50-mum increments, with the results then normalized per unit area of marrow tissue. The distances separating hematopoietic CD34(+) cells from vessels were also tallied. RESULTS: Hematopoietic CD34(+) cells were found to exist along a linear spatial gradient with a maximal areal concentration localized within the first 50 mum of the bone surfaces. An exponential spatial concentration gradient was found in the concentration of blood vessel fragments within the images. Distances between hematopoietic CD34(+) cells and blood vessels exhibited a lognormal distribution indicating a shared spatial niche. CONCLUSION: Study results confirm that the spatial gradient of hematopoietic stem and progenitor cells previously measured in mouse femur is also present within human cancellous bone. The dosimetric implication of these results may be significant for those scenarios in which the absorbed dose itself is nonuniformly delivered across the marrow tissues, as would be the case for a low-energy beta- or alpha-particle emitter localized on the bone surfaces. PMID- 17401105 TI - Biodistribution, PET, and radiation dosimetry estimates of HSV-tk gene expression imaging agent 1-(2'-Deoxy-2'-18F-Fluoro-beta-D-arabinofuranosyl)-5-iodouracil in normal dogs. AB - FIAU is of interest as a potential reporter probe to monitor herpes simplex virus thymidine kinase (HSV-tk) gene expression and bacterial infections. This study investigates the biodistribution, metabolism, and DNA uptake of 1-(2'-deoxy-2' (18)F-fluoro-beta-d-arabinofuranosyl)-5-iodouracil ((18)F-FIAU) in normal dogs. METHODS: Four normal dogs were intravenously administered (18)F-FIAU. A dynamic PET scan was performed for 60 min over the upper abdomen; this was followed by a whole-body scan for a total of 150 min on 3 dogs. The fourth dog was not scanned and was euthanized at 60 min. Blood and urine samples were collected at stipulated time intervals and analyzed by high-performance liquid chromatography to evaluate tracer clearance and metabolism. Tissue samples collected from various organs were analyzed to evaluate tracer uptake and DNA incorporation. Dynamic accumulation of the tracer in different organs was derived from reconstructed PET images. Nondecay-corrected time-activity curves were used for residence time calculation and absorbed dose estimation. RESULTS: At 60 min after injection, unmetabolized FIAU radioactivity in blood and urine samples was greater than 78% and 63%, respectively, demonstrating resistance to metabolism. The tissue-to-muscle ratio derived from image and tissue analysis showed a slightly higher uptake in proliferating organs (mean tissue-to-muscle values: small intestine, 1.97; marrow, 1.70) compared with nonproliferative organs (heart, 1.07; lung, 1.06). A high concentration of activity was seen in the bile (mean, 23.02), demonstrating hepatobiliary excretion of the tracer. Extraction analysis of tissue samples showed that >62% of the activity in the small intestine, 74% in marrow, and <21% in heart, liver, and muscle was incorporated into DNA. CONCLUSION: These results demonstrate that FIAU is resistant to metabolism and moderately incorporates into DNA in proliferating tissues. These results suggest that incorporation into the DNA of normal tissues may need to be considered when FIAU is used to track reporter gene activity. Studies in humans are needed to determine whether imaging properties differ in patients and are altered as a result of metabolism changes affected by gene therapies. PMID- 17401106 TI - Recent advances in SPECT imaging. AB - SPECT is a rapidly changing field, and the past several years have produced new developments in both hardware technology and image-processing algorithms. At the component level there have been improvements in scintillators and photon transducers as well as a greater availability of semiconductor technology. These devices permit the fabrication of smaller and more compact systems that can be customized for particular applications. New clinical devices include high-count sensitivity cardiac SPECT systems that do not use conventional collimation and the introduction of diagnostic-quality hybrid SPECT/CT systems. While there has been steady progress with reconstruction algorithms, exciting new processing algorithms have become commercially available that promise to provide substantial reductions in SPECT acquisition time without sacrificing diagnostic quality. Preclinical small-animal SPECT systems have become a major focus in nuclear medicine. These systems have pushed the limits of SPECT into the submillimeter range, making them valuable molecular imaging tools capable of providing information unavailable from other modalities. PMID- 17401107 TI - Re: the so-called stunning of thyroid tissue. PMID- 17401109 TI - Perilipin regulates the thermogenic actions of norepinephrine in brown adipose tissue. AB - In response to cold, norepinephrine (NE)-induced triacylglycerol hydrolysis (lipolysis) in adipocytes of brown adipose tissue (BAT) provides fatty acid substrates to mitochondria for heat generation (adaptive thermogenesis). NE induced lipolysis is mediated by protein kinase A (PKA)-dependent phosphorylation of perilipin, a lipid droplet-associated protein that is the major regulator of lipolysis. We investigated the role of perilipin PKA phosphorylation in BAT NE stimulated thermogenesis using a novel mouse model in which a mutant form of perilipin, lacking all six PKA phosphorylation sites, is expressed in adipocytes of perilipin knockout (Peri KO) mice. Here, we show that despite a normal mitochondrial respiratory capacity, NE-induced lipolysis is abrogated in the interscapular brown adipose tissue (IBAT) of these mice. This lipolytic constraint is accompanied by a dramatic blunting ( approximately 70%) of the in vivo thermal response to NE. Thus, in the presence of perilipin, PKA-mediated perilipin phosphorylation is essential for NE-dependent lipolysis and full adaptive thermogenesis in BAT. In IBAT of Peri KO mice, increased basal lipolysis attributable to the absence of perilipin is sufficient to support a rapid NE stimulated temperature increase ( approximately 3.0 degrees C) comparable to that in wild-type mice. This observation suggests that one or more NE-dependent mechanism downstream of perilipin phosphorylation is required to initiate and/or sustain the IBAT thermal response. PMID- 17401110 TI - Further biochemical characterization of human pancreatic lipase-related protein 2 expressed in yeast cells. AB - Recombinant human pancreatic lipase-related protein 2 (rHPLRP2) was produced in the protease A-deficient yeast Pichia pastoris. A major protein with a molecular mass of 50 kDa was purified from the culture medium using SP-Sepharose and Mono Q chromatography. The protein was found to be highly sensitive to the proteolytic cleavage of a peptide bond in the lid domain. The proteolytic cleavage process occurring in the lid affected both the lipase and phospholipase activities of rHPLRP2. The substrate specificity of the nonproteolyzed rHPLRP2 was investigated using pH-stat and monomolecular film techniques and various substrates (glycerides, phospholipids, and galactolipids). All of the enzyme activities were maximum at alkaline pH values and decreased in the pH 5-7 range corresponding to the physiological conditions occurring in the duodenum. rHPLRP2 was found to act preferentially on substrates forming small aggregates in solution (monoglycerides, egg phosphatidylcholine, and galactolipids) rather than on emulsified substrates such as triolein and diolein. The activity of rHPLRP2 on monogalactosyldiglyceride and digalactosyldiglyceride monomolecular films was determined and compared with that of guinea pig pancreatic lipase-related protein 2, which shows a large deletion in the lid domain. The presence of a full-length lid domain in rHPLRP2 makes it possible for enzyme activity to occur at higher surface pressures. The finding that the inhibition of nonproteolyzed rHPLRP2 by tetrahydrolipstatin and diethyl-p-nitrophenyl phosphate does not involve any bile salt requirements suggests that the rHPLRP2 lid adopts an open conformation in aqueous media. PMID- 17401111 TI - Avian apolipoprotein A-V binds to LDL receptor gene family members. AB - Apolipoprotein A-V (apoA-V) affects plasma triglyceride (TG) levels; however, the properties of apoA-V that mediate its action(s) are still incompletely understood. It is unclear how apoA-V, whose plasma concentration is extremely low, can affect the pronounced TG differences observed in individuals with various apoA-V dysfunctions. To gain novel insights into apoA-V biology, we expanded our previous studies in the chicken to this apolipoprotein. First, we characterized the first avian apoA-V, revealing its expression not only in liver and small intestine but also in brain, kidney, and ovarian follicles and showing its presence in the circulation. Second, we demonstrate directly that galline apoA-V binds to the major LDL receptor family member (LR) of the laying hen and that this interaction does not depend on the association of the apolipoprotein with lipid or lipoproteins. We propose that a direct interaction with LRs may represent a novel, additional mechanism for the modulation of TG levels by apoA V. PMID- 17401112 TI - The role of GW/P-bodies in RNA processing and silencing. AB - GW bodies, also known as mammalian P-bodies, are cytoplasmic foci involved in the post-transcriptional regulation of eukaryotic gene expression. Recently, GW bodies have been linked to RNA interference and demonstrated to be important for short-interfering-RNA- and microRNA-mediated mRNA decay and translational repression. Evidence indicates that both passenger and guide strands of short interfering RNA duplexes can localize to GW bodies, thereby indicating that RNA induced silencing complexes may be activated within these cytoplasmic centers. Formation of GW bodies appears to depend on both specific protein factors and RNA, in particular, microRNA. Work over the past few years has significantly increased our understanding of the biology of GW bodies, revealing that they are specialized cell components that spatially regulate mRNA turnover in various biological processes. The formation of GW bodies appears to depend on both specific protein factors and RNA, in particular, microRNA. Here, we propose a working model for GW body assembly in terms of its relationship to RNA interference. In this process, one or more heteromeric protein complexes accumulate in successive steps into larger ribonucleoprotein structures. PMID- 17401113 TI - Analysis of actin filament network organization in lamellipodia by comparing experimental and simulated images. AB - Protrusion of lamellipodia during cell migration depends on the assembly of actin network. The assembly mechanism, based on dendritic filament branching, has been investigated in reconstituted in vitro systems, but little is known about the dynamical and structural properties of the actin network in the lamellipodia of migrating cells. The length and orientation of filaments are difficult to measure directly in either optical or electron microscopy images because of the high filament density and overlapping of individual filaments. Here, we use the non uniformity of optical images of the lamellipodia to extract information about the structural and dynamical properties of the underlying actin network. To determine the relationship between the image features and the properties of the network, we performed simulations of actin network assembly, based on the hypothesis of stochastic branching and capping of filaments, and produced computed ;fluorescence' and ;electron microscopy' images of the simulated network. By varying simulation parameters, in particular the actin filament density, length and orientation, we closely reproduced the contrast and the characteristic diagonal criss-cross pattern observed in the experimental optical images. Thus, matching the images of the simulated network to the experimental images allowed us to estimate parameters of actin filament network in lamellipodia. PMID- 17401114 TI - Osmoregulation in elephant fish Callorhinchus milii (Holocephali), with special reference to the rectal gland. AB - Osmoregulatory mechanisms in holocephalan fishes are poorly understood except that these fish are known to conduct urea-based osmoregulation as in elasmobranchs. We, therefore, examined changes in plasma parameters of elephant fish Callorhinchus milii, after gradual transfer to concentrated (120%) or diluted (80%) seawater (SW). In control fish, plasma Na and urea concentrations were about 300 mmol l(-1) and 450 mmol l(-1), respectively. These values were equivalent to those of sharks and rays, but the plasma urea concentration of elephant fish was considerably higher than that reported for chimaeras, another holocephalan. After transfer to 120% SW, plasma osmolality, urea and ion concentrations were increased, whereas transfer to 80% SW resulted in a fall in these parameters. The rises in ion concentrations were notable after transfer to 120% SW, whereas urea concentration decreased predominantly following transfer to 80% SW. In elephant fish, we could not find a discrete rectal gland. Instead, approximately 10 tubular structures were located in the wall of post-valvular intestine. Each tubular structure was composed of a putative salt-secreting component consisting of a single-layered columnar epithelium, which was stained with an anti-Na(+),K(+)-ATPase serum. Furthermore, Na(+),K(+)-ATPase activity in the tubular structures was significantly increased after acute transfer of fish to concentrated SW (115%). These results suggest that the tubular structures are a rectal gland equivalent, functioning as a salt-secreting organ. Since the rectal gland of elephant fish is well developed compared to that of Southern chimaera, the salt-secreting ability may be higher in elephant fish than chimaeras, which may account for the lower plasma NaCl concentration in elephant fish compared to other chimaeras. Since elephant fish have also attracted attention from a viewpoint of genome science, the availability of fish for physiological studies will make this species an excellent model in holocephalan fish group. PMID- 17401115 TI - Social domination increases neuronal survival in the brain of juvenile crayfish Procambarus clarkii. AB - Olfactory cues are among the sensory inputs that crayfish use in establishing dominance hierarchies. Throughout their lives, new neurons are continuously added into brain cell clusters 9 and 10, which contain somata of olfactory local and projection interneurons, respectively. Using markers for DNA synthesis (bromodeoxyuridine) and mitosis (phospho-histone-3), we tested juvenile crayfish (Procambarus clarkii) to examine effects of pairwise social experience on proliferation and survival of cells in these brain regions. Proliferating and mitotic cells appeared within restricted neurogenic areas in both clusters and in ;tails' extending from them. These tails, embedded in tubulin-positive strands, are linked by a patch of cells. Neither cell proliferation nor mitotic activity was affected by social dominance. Cell survival of neuronal precursors was affected by dominance: compared to dominants, subordinates had fewer newborn cells surviving in cluster 9 after 14 days of social experience. Social experience also affected body growth rate, but the effect of social experience on neurogenesis remained when differences in body growth rate were statistically controlled. We conclude that social domination enhances survival of new olfactory interneuronal precursors compared to social subordination but not compared to social isolation. PMID- 17401116 TI - What comes first, the zebra finch or the egg: temperature-dependent reproductive, physiological and behavioural plasticity in egg-laying zebra finches. AB - Avian reproduction is generally timed to synchronize chick-rearing with periods of increased food abundance. Consequently, the energetically demanding period of egg production may coincide with periods of lower food availability, fluctuating temperature and more unstable weather. Little is known about the physiological mechanisms underlying temperature-induced variation in egg production. We therefore examined the influence of low ambient temperature (7 degrees C vs 21 degrees C) on reproductive output (e.g. egg mass, clutch size, laying interval, laying rate), daily food consumption and lipid variables in zebra finches Taeniopygia guttata. When faced with egg production at 7 degrees C, laying zebra finches increased energy intake by 12.67 kJ day(-1), and were thus able to maintain body condition (e.g. body mass, fat and muscle score) and circulating triacylglyceride at levels comparable to those at 21 degrees C. However, when producing eggs at 7 degrees C, females took longer to initiate egg laying (6.5 vs 6.1 days at 21 degrees C), and ultimately laid fewer eggs (5.5 vs 6.0 eggs) at a slower rate (0.90 eggs day(-1) vs 0.95 eggs day(-1)). These temperature-related declines in reproductive output were accompanied by decreases in modal (from 36.6 at 21 degrees C to 24.3 nm at 7 degrees C) and median very low density lipoprotein (VLDL) particle diameter (from 29.6 to 26.4 nm) and in the proportion of VLDL particles that were capable of passing through the pores in the ovary to access the developing ovarian follicles (i.e. particles with diameters between 25 and 44 nm; from 45.90% to 32.55%). However, variation in reproductive output was not related to any static concentration or structural measure of VLDL. Therefore, other temperature-dependent mechanisms must be involved in the physiological processes that regulate reproductive output of passerine birds at low ambient temperatures. PMID- 17401117 TI - Osmoregulation, ionoregulation and acid-base regulation by the gastrointestinal tract after feeding in the elasmobranch (Squalus acanthias). AB - In order to study the physiological consequences of voluntary feeding in the gastrointestinal tract of a ureotelic marine elasmobranch, dogfish (fasted for 96 h) were sampled at various times up to 360 h after consuming a 5-6% ration of teleost fish (hake) under natural feeding conditions. Digestion and absorption were completed between 120 and 360 h post-feeding. The tissue masses of different segments of the gastrointestinal tract increased and decreased markedly as the chyme moved through, mainly because of fluid engorgement rather than hyperplasia. In fasted dogfish, the cardiac and pyloric stomachs contained only small volumes of highly acidic fluid (pH 1.77+/-1.12, 2.05+/-0.08) similar in composition to seawater. Feeding resulted in gastric pHs of 3.20+/-0.31 and 3.95+/-0.40 at 6 h, followed by slow declines through 60 h. An alkaline tide in the blood also occurred at 6 h. In the face of large changing masses of highly acidic chyme in the stomachs, the pH (6.50+/-0.10), ionic composition and volume of chyme in the intestine (spiral valve) were precisely regulated from 6 to 60 h post-feeding at very different values from those in the stomachs, and intestinal HCO3(-) remained low (5.12+/-0.83 mmol l(-1)). The colon was usually empty and its pH constant at 7.20+/-0.16 at all times. Despite the ingestion of strongly hypo-osmotic teleost tissue, the osmolality of the chyme remained in equilibrium with that of the blood plasma in all segments at all times after feeding. Much of the osmotic equilibration was because of the secretion of urea into the chyme, particularly in the intestine. After feeding, gastric fluid concentrations of Na(+) and Mg(2+) declined, K(+) and Ca(2+) increased, whereas Cl(-) exhibited little change, indicating that additional drinking of seawater was minimal. Na(+), K(+), water and especially Cl(-) were absorbed in the intestine, whereas Mg(2+) and Ca(2+) were largely excluded. Our results illustrate the complex integration of digestive and ionoregulatory function in the elasmobranch digestive tract, and marked differences from the teleost pattern. PMID- 17401118 TI - Spectral properties of identified polarized-light sensitive interneurons in the brain of the desert locust Schistocerca gregaria. AB - Many migrating animals employ a celestial compass mechanism for spatial navigation. Behavioral experiments in bees and ants have shown that sun compass navigation may rely on the spectral gradient in the sky as well as on the pattern of sky polarization. While polarized-light sensitive interneurons (POL neurons) have been identified in the brain of several insect species, there are at present no data on the neural basis of coding the spectral gradient of the sky. In the present study we have analyzed the chromatic properties of two identified POL neurons in the brain of the desert locust. Both neurons, termed TuTu1 and LoTu1, arborize in the anterior optic tubercle and respond to unpolarized light as well as to polarized light. We show here that the polarized-light response of both types of neuron relies on blue-sensitive photoreceptors. Responses to unpolarized light depended on stimulus position and wavelength. Dorsal unpolarized blue light inhibited the neurons, while stimulation from the ipsilateral side resulted in opponent responses to UV light and green light. While LoTu1 was inhibited by UV light and was excited by green light, one subtype of TuTu1 was excited by UV and inhibited by green light. In LoTu1 the sensitivity to polarized light was at least 2 log units higher than the response to unpolarized light stimuli. Taken together, the spatial and chromatic properties of the neurons may be suited to signal azimuthal directions based on a combination of the spectral gradient and the polarization pattern of the sky. PMID- 17401119 TI - The aerodynamic benefit of wing-wing interaction depends on stroke trajectory in flapping insect wings. AB - Flying insects may enhance their flight force production by contralateral wing interaction during dorsal stroke reversal ('clap-and-fling'). In this study, we explored the forces and moments due to clap-and-fling at various wing tip trajectories, employing a dynamically scaled electromechanical flapping device. The 17 tested bio-inspired kinematic patterns were identical in stroke amplitude, stroke frequency and angle of attack with respect to the horizontal stroke plane but varied in heaving motion. Clap-and-fling induced vertical force augmentation significantly decreased with increasing vertical force production averaged over the entire stroke cycle, whereas total force augmentation was independent from changes in force produced by a single wing. Vertical force augmentation was also largely independent of forces produced due to wing rotation at the stroke reversals, the sum of rotational circulation and wake capture force. We obtained maximum (17.4%) and minimum (1.4%) vertical force augmentation in two types of figure-eight stroke kinematics whereby rate and direction of heaving motion during fling may explain 58% of the variance in vertical force augmentation. This finding suggests that vertical wing motion distinctly alters the flow regime at the beginning of the downstroke. Using an analytical model, we determined pitching moments acting on an imaginary body of the flapping device from the measured time course of forces, the changes in length of the force vector's moment arm, the position of the centre of mass and body angle. The data show that pitching moments are largely independent from mean vertical force; however, clap and-fling reinforces mean pitching moments by approximately 21%, compared to the moments produced by a single flapping wing. Pitching moments due to clap-and fling significantly increase with increasing vertical force augmentation and produce nose-down moments in most of the tested patterns. The analytical model, however, shows that algebraic sign and magnitude of these moments may vary distinctly depending on both body angle and the distance between the wing hinge and the animal's centre of mass. Altogether, the data suggest that the benefit of clap-and-fling wing beat for vertical force enhancement and pitch balance may change with changing heaving motion and thus wing tip trajectory during manoeuvring flight. We hypothesize that these dependencies may have shaped the evolution of wing kinematics in insects that are limited by aerodynamic lift rather than by mechanical power of their flight musculature. PMID- 17401120 TI - Mechanics of cutting maneuvers by ostriches (Struthio camelus). AB - We studied the strategies used by cursorial bipeds (ostriches) to maneuver during running. Eight ostriches were induced to run along a trackway and execute turns. Ground reaction forces and three-dimensional kinematics of the body and leg joints were simultaneously recorded, allowing calculation of joint angles and quasi-static net joint torques. Sidesteps, where the leg on the outside of the turn changes the movement direction, and crossovers using the inside leg, occurred with nearly equal frequency. Ostriches executed maneuvers using a simple control strategy that required minimal changes to leg kinematics or net torque production at individual joints. Although ostriches did use acceleration or braking forces to control body rotation, their morphology allowed for both crossovers and sidesteps to be accomplished with minimal net acceleratory/braking force production. Moreover, body roll and ab/adduction of the leg shifted the foot position away from the turn direction, reducing the acceleratory/braking forces required to prevent under- or over-rotation and aligning the leg with the ground reaction force. PMID- 17401121 TI - Annual changes in body mass and resting metabolism in captive barnacle geese (Branta leucopsis): the importance of wing moult. AB - Many different physiological changes have been observed in wild waterfowl during the flightless stage of wing moult, including a loss of body mass. We aimed to determine whether captive barnacle geese (Branta leucopsis) would show the characteristic decrease in body mass during their wing moult, even though they had unlimited and unrestricted access to food. Fourteen captive geese were weighed at 1-2-week intervals for two complete years. During the flightless period of the moult, body mass decreased by approximately 25% from the pre-moult value. To understand the basis of this change, the rate of oxygen consumption was measured during daytime and nighttime at six points in the second year, and at three points (before, during and after wing moult) behavioural observations were made. Measurements of the rate of oxygen consumption showed an 80% increase above that of the nonmoulting periods of the year. We propose that metabolism was increased during moult because of the cost of feather synthesis. Although food was available, the captive birds chose not to forage and instead increased the proportion of time spent resting. It is likely that this behaviour in response to wing moult is a strategy to avoid predation in the wild. Thus, the innate nature of this behaviour has potential survival value for wild birds of this species. We conclude that the increase in metabolism led to the use of endogenous energy reserves because the birds reduced rather than increased their food intake rates, and as a result, the barnacle geese lost body mass during wing moult. PMID- 17401122 TI - The effect of decoupling olfactory and visual stimuli on the foraging behavior of Manduca sexta. AB - Within an appetitive context, Manduca sexta, a nectivorous nocturnal hawkmoth, can be attracted by a range of stimuli including floral volatiles and visual display, carbon dioxide and water vapor. Several studies on this and other flower visiting insects have shown how olfactory and visual stimulation play (or do not play) a role in attraction and feeding. Nevertheless, these studies have consistently manipulated stimuli in a 'presence-absence' manner. Here, we experimentally decoupled the presentation of both stimuli spatially and temporally in a wind tunnel, rather than entirely eliminating either one, and found that the decision-making process based on these stimuli is more flexible and complex than previously asserted. Manduca sexta was most responsive when both cues were present and emanated from the same source. When stimuli were spatially separated, responsiveness levels were comparable to those elicited by a single stimulus. However, transient olfactory stimulation either before or after visually guided approach (temporal decoupling) enhanced responsiveness to an odorless visual target. Additionally, searching times were increased by either a transient olfactory stimulation before take-off or by having the flower model spatially separated from the odor source tracked by the moths. Finally, in a dual choice experiment, moths showed a strong bias for the visual display over the odor plume, suggesting the former to be the ultimate indicator of a nectar source. Our manipulation of floral cues shows that the feeding behavior of M. sexta, and probably of other nectivorous insects, is based not only on the sensory stimulation per se but also on the temporal and spatial pattern in which these stimuli are perceived. PMID- 17401123 TI - Pregnancy block by MHC class I peptides is mediated via the production of inositol 1,4,5-trisphosphate in the mouse vomeronasal organ. AB - The vomeronasal organ (VNO) has evolved to link an animal's behavior to its environment in a highly species-specific fashion. In mice, it is thought to be the primary sensory system responsible for the detection of pheromones. Pheromones regulate a variety of responses including mate recognition in the context of selective pregnancy failure. MHC (major histocompatibility complex) class I peptides have been identified as compounds that elicit the pregnancy block effect via the VNO. However, the transduction cascade of these molecules is unknown and it is not known if the production of these compounds are androgen dependent. By using male urine and MHC peptides, we show that female mice treated with MHC peptides (in urine or PBS) and urine from castrated males or juvenile mice of different haplotypes respond to the Bruce Effect paradigm in a manner equivalent to female mice exposed to whole urine. In addition to providing new evidence that urine from castrated or juvenile males and MHC peptides can induce pregnancy block, we show correlation of the effect with an increase in inositol 1,4,5-trisphosphate. PMID- 17401124 TI - Take-off and landing forces and the evolution of controlled gliding in northern flying squirrels Glaucomys sabrinus. AB - Flying squirrels are well known for their ability to glide between trees at the top of a forest canopy. We present experimental performance and behavioural evidence that flight in flying squirrels may have evolved out of a need to control landing forces. Northern flying squirrels were filmed jumping from a horizontal branch to a much larger vertical pole. These were both slightly compliant (less than 1.9 mm N(-1)), and instrumented using strain gauges so that forces could be measured. Take-off and landing forces were both positively correlated with horizontal range between 0.5 and 2.5 m (r=0.355 and r=0.811, respectively, P<0.05), but not significantly different to each other at each range tested. Take-off forces ranged from 1 to 10 bodyweights, and landing forces were between 3 and 10 bodyweights. Glide angles increased rapidly with horizontal range, approaching 45 degrees at 3 m, above which they gradually decreased, suggesting that northern flying squirrels are optimised for long distance travel. We show that northern flying squirrels initiate full gliding posture at ranges of less than 1 m, without landing any higher than an equivalent ballistic projectile. However, this gliding posture enables them to pitch upwards, potentially stalling the wing, and spreads the landing reaction force over all four extended limbs. At steeper approach angles of close to 45 degrees , flying squirrels were unable to pitch up sufficiently and landed forelimbs first, consequently sustaining higher impact forces. We investigate four hypotheses to explain the origin of flight in these animals and conclude that the need to reduce landing impact forces was most likely to have stimulated the development of aerial control in flying squirrels. PMID- 17401125 TI - Regulation of feeding behaviour and locomotor activity by takeout in Drosophila. AB - The hormonal regulation of feeding behaviour is well known in vertebrates, whereas it remains poorly understood in insects. Here, we report that the takeout gene is an essential component of nutritional homeostasis in Drosophila. takeout encodes a putative juvenile hormone (JH) binding protein and has been described as a link between circadian rhythm and feeding behaviour. However, the physiological role of takeout and its putative link to JH remain unknown. In this study, we show that takeout (to(1)) flies failed to adapt their food intake according to food availability and that most defects could be genetically rescued. When food is abundant, to(1) are hyperphagic, yielding to hypertrophy of the fat body. When food reappears after a starvation period, to(1) flies do not increase their food intake as much as wild-type flies. This defect in food intake regulation is partly based on the action of Takeout on taste neurons, because the sensitivity of to(1) gustatory neurons to sugars does not increase after starvation, as in wild-type neurons. This lack of regulation is also evident at the locomotor activity, which normally increases during starvation, a behaviour related to food foraging. In addition, to(1) flies lack sexual dimorphism of locomotor activity, which has previously been linked to the JH circulating level. Moreover, application of the JH analog methoprene rescues the phenotype. These results suggest that takeout plays a central role as a feeding regulator and may act by modulating the circulating JH level. PMID- 17401126 TI - Parameters influencing the dissolved oxygen in the boundary layer of rainbow trout (Oncorhynchus mykiss) embryos and larvae. AB - We investigated the influence of oxygen demand (developmental stage) and supply (hypoxia, water flow rate, the chorion and body movements) on the oxygen concentration within the boundary layer next to the chorion of embryos or skin of larvae of rainbow trout (Oncorhynchus mykiss). Oxygen microelectrodes were used to measure dissolved oxygen (DO) within the boundary layer of trout embryos and larvae. As the embryos and larvae developed, the DO gradient and the thickness of the boundary layer increased. The DO concentration within the boundary layer next to the chorion or skin surface decreased as the DO concentration in the free stream water decreased. A decrease in water flow rate increased the magnitude of the gradient and thickness of the boundary layer. In normoxia, the DO in the perivitelline fluid inside the chorion was 16+/-3.0% saturation at 31 days post fertilization, indicating that the chorion was a significant barrier to oxygen diffusion. The number of body movements did not change when embryos were exposed to hypoxia before hatching, but after hatching, hypoxia resulted in a decrease in body movements of the larvae. Taken together, our data indicate that the oxygen boundary layer around trout embryos and larvae depends on both the oxygen demand and supply. The factors that significantly impacted boundary layer oxygen were developmental stage, free-stream oxygen levels, water flow rate, and the presence of the chorion. PMID- 17401127 TI - Just drop it and run: the effect of limb autotomy on running distance and locomotion energetics of field crickets (Gryllus bimaculatus). AB - This is the first study to examine the direct metabolic costs of autotomy, the voluntary shedding of an appendage as an escape mechanism, in invertebrates. We investigated the effects of limb autotomy upon endurance and metabolic cost of locomotion in the field cricket Gryllus bimaculatus. Compared with control (intact) crickets, animals that had autotomised a single hindlimb were slower, stopped more often, moved a shorter distance and expended more energy doing so. Both the cost of locomotion (COT) and minimal cost of locomotion (MCOT) were significantly higher for autotomised animals. We compare these data with locomotion energetics of 36 other invertebrate species, and discuss the results in terms of the biomechanics of walking in crickets. PMID- 17401128 TI - Neuroprotection from secondary injury by polyethylene glycol requires its internalization. AB - Polyethylene glycol (PEG) is well known to both fuse and repair cell membranes. This capability has been exploited for such diverse usages as the construction of hybridomas and as a reparative agent following neurotrauma. The latter development has proceeded through preclinical testing in cases of naturally induced paraplegia in dogs. The mechanisms of action of polymer-mediated neurorepair/neuroprotection are still under investigation. It is likely that the unique interaction of hydrophilic polymers with the mechanical properties of cell membranes in concert with an ability to interfere with mechanisms of secondary injury such as the production of highly reactive oxygen species (ROS or ;free radicals') is the basis for neuroprotection by polymers. Here we provide further evidence that the ability of PEG to reduce or limit secondary injury and/or lipid peroxidation (LPO) of membranes requires entry of PEG into the cytosol, further suggesting a physical interaction with the membranes of organelles such as mitochondria as the initial event leading to neurorepair/neuroprotection. We have evaluated this relationship in vitro using acrolein, a potent endogenous toxin that is a product of LPO. Acrolein can pass through cell membranes with ease, inducing progressive LPO in ;bystander' cells, and the production of even more acrolein by inducing its own production. Immediate application of PEG (10 mmol l( 1), 2000 Da) to poisoned neurons in vitro was unable to rescue them from necrosis and death. Furthermore, three-dimensional confocal microscopy of fluorescently decorated PEG shows that it does not enter these cells for up to 2 h after application. By this time the mechanisms of necrosis are likely irreversible. Additionally, severe oxygen and or glucose deprivation of spinal cord white matter in vitro also initiates LPO. Addition of potent free radical scavengers such as ascorbic acid or superoxide dismutase (SOD) is able to interfere with this process, but PEG is not. Taken together, these data are consistent with the hypothesis that PEG is able to rescue mechanically damaged cells, based on a restructuring of the damaged plasmalemma. Furthermore, in compromised cells with an intact cell membrane, PEG must first gain access to the cytosol where this same capability may be useful in restoring the integrity of cellular organelles such as mitochondria, though the intracellular concentration of the polymer must be significant relative to the concentration of toxins produced by LPO in order to rescue the cell. PMID- 17401129 TI - A vacuolar-type H+-ATPase and a Na+/H+ exchanger contribute to intracellular pH regulation in cockroach salivary ducts. AB - Cells of the dopaminergically innervated salivary ducts in the cockroach Periplaneta americana have a vacuolar-type H(+)-ATPase (V-ATPase) of unknown function in their apical membrane. We have studied whether dopamine affects intracellular pH (pH(i)) in duct cells and whether and to what extent the apical V-ATPase contributes to pH(i) regulation. pH(i) measurements with double barrelled pH-sensitive microelectrodes and the fluorescent dye BCECF have revealed: (1) the steady-state pH(i) is 7.3+/-0.1; (2) dopamine induces a dose dependent acidification up to pH 6.9+/-0.1 at 1 micromol l(-1) dopamine, EC(50) at 30 nmol l(-1) dopamine; (3) V-ATPase inhibition with concanamycin A or Na(+) free physiological saline (PS) does not affect the steady-state pH(i); (4) concanamycin A, Na(+) -free PS and Na(+)/H(+) exchange inhibition with 5-(N-ethyl N-isopropyl)-amiloride (EIPA) each reduce the rate of pH(i) recovery from a dopamine-induced acidification or an acidification induced by an NH(4)Cl pulse; (5) pH(i) recovery after NH(4)Cl-induced acidification is almost completely blocked by concanamycin A in Na(+)-free PS or by concanamycin A applied together with EIPA; (6) pH(i) recovery after dopamine-induced acidification is also completely blocked by concanamycin A in Na(+)-free PS but only partially blocked by concanamycin A applied together with EIPA. We therefore conclude that the apical V-ATPase and a basolateral Na(+)/H(+) exchange play a minor role in steady state pH(i) regulation but contribute both to H(+) extrusion after an acute dopamine- or NH(4)Cl-induced acid load. PMID- 17401130 TI - The urinary bladder as a physiological reservoir that moderates dehydration in a large desert lizard, the Gila monster Heloderma suspectum. AB - Animals inhabiting xeric environments use a variety of behavioral and physiological strategies to balance water budgets. We studied the potential contribution of the urinary bladder to osmoregulation in a large desert lizard, the Gila monster Heloderma suspectum. Here we present results of a series of in vivo laboratory experiments which tested the hypothesis that the Gila monster urinary bladder serves as a physiological reservoir, as in amphibians and chelonians, providing water that buffers increases in plasma osmolality when food and water are unavailable. Adult Gila monsters absorbed water from the urinary bladder into circulation and absorption of water from the urinary bladder and drinking water provided similar osmoregulatory benefits within 24 h, although drinking water provided a more immediate osmotic benefit. During food and water deprivation, plasma osmolality increased 2.5 times faster in lizards with an empty urinary bladder compared with those with a full bladder. During rehydration, stereotyped binge drinking behavior increased body mass nearly 22%, which resulted in a 24% reduction in plasma osmolality and a substantial increase in bladder water within 24 h. These results support our hypothesis and demonstrate for the first time in an adult lizard that the urinary bladder can function as a long-term physiological water reservoir. This trait can provide a critical benefit to osmoregulation during the 2- to 3-month summer dry season characteristic of the deserts that Gila monsters inhabit. PMID- 17401131 TI - The role of calcium and magnesium in the concrete tubes of the sandcastle worm. AB - Sandcastle worms Phragmatopoma californica build mound-like reefs by sticking together large numbers of sand grains with cement secreted from the building organ. The cement consists of protein plus substantial amounts of calcium and magnesium, which are not invested in any mineral form. This study examined the effect of calcium and magnesium depletion on the structural and mechanical properties of the cement. Divalent ion removal by chelating with EDTA led to a partial collapse of cement architecture and cement dislodgement from silica surfaces. Mechanical properties examined were sand grain pull-out force, tube resistance to compression and cement adhesive force. EDTA treatment reduced sand grain pull-out forces by 60% and tube compressive strength by 50% relative to controls. EDTA lowered both the maximal adhesive force and energy dissipation of cement by up to an order of magnitude. The adhesiveness of calcium- and magnesium depleted cement could not be restored by re-exposure to the ions. The results suggest that divalent ions play a complex and multifunctional role in maintaining the structure and stickiness of Phragmatopoma cement. PMID- 17401132 TI - The impact of the NICE appraisal consultation document on anti-TNF therapy for ankylosing spondylitis. PMID- 17401133 TI - Acculturation and the prevalence of pain amongst South Asian minority ethnic groups in the UK. AB - BACKGROUND: Musculoskeletal pain is reported more commonly by South Asians in the UK than by white Europeans. This may result from a variety of factors, including cultural differences, and thus we investigated the extent to which differences in the prevalence of pain within the South Asian population could be explained by differences in acculturation (the extent to which immigrants take on the culture of their host population). METHODS: Nine hundred and thirty-three Europeans and 1914 South Asian (1165 Indian, 401 Pakistani and 348 Bangladeshi) subjects were recruited from the age-sex registers of 13 general practices in areas with high densities of South Asian populations (Bolton, Oldham, Ashton-under-Lyne and Birmingham). A 28-item acculturation scale was developed, based, on aspects including use of language, clothing style, and use of own-culture media. Principle component analysis generated a score (range 0-100), which was validated against constructs expected to relate to acculturation, such as years of full time education and time spent in the UK. The presence of widespread pain was assessed by the answer to the question 'Have you suffered from pain all over the body in the past month?' RESULTS: Widespread pain was more common in all three South Asian ethnic groups than in the white Europeans [odds ratio (OR) = 3.7, 95% confidence interval (CI) 2.9-4.9], with this increase ranging from 2.7 to 5.8 in the different South Asian subgroups. There was a similar increase in consultation rates for pain. Within the South Asians, pooling all three groups, there was a strong negative association between acculturation score and widespread pain, which remained after adjusting for age and sex: [OR (95% CI) per standard deviation decrease in acculturation score -1.2 (1.0-1.3)]. Adjusting for acculturation accounted for some, but not all, of the differences between the ethnic groups in the prevalence of widespread pain: OR 2.0 (95% CI 1.4-3.0). CONCLUSIONS: Widespread pain is more commonly reported in South Asians though there are interesting differences within the South Asian community. Lower acculturation has a strong influence on the reporting of pain, but cannot explain all of the difference between South Asian and European populations. PMID- 17401134 TI - Current role of glucosamine in the treatment of osteoarthritis. AB - OBJECTIVES: To evaluate the interest of using the various preparations of glucosamine for symptomatic and structural management of osteoarthritis (OA). METHODS: A critical analysis of the literature based on an exhaustive search (Medline, PubMed and manual search within the bibliography of retrieved manuscripts) from 1980 to 2005. RESULTS: Despite multiple controlled clinical trials of the use of glucosamine in OA (mainly of the knee), controversy on efficacy related to symptomatic improvement continues. Differences in results originate from the differences in products, study design and study populations. Symptomatic efficacy described in multiple studies performed with glucosamine sulphate (GS) support continued consideration in the OA therapeutic armamentarium. The most compelling evidence of a potential for inhibiting the progression of OA is also obtain with GS. CONCLUSIONS: GS has shown positive effects on symptomatic and structural outcomes of knee OA. These results should not be extrapolated to other glucosamine salts [hydrochloride or preparations (over-the-counter or food supplements)] in which no warranty exists about content, pharmacokinetics and pharmacodynamics of the tablets. PMID- 17401135 TI - Antisense expression of 3-oxoacyl-ACP reductase affects whole plant productivity and causes collateral changes in activity of fatty acid synthase components. AB - Brassica napus cv Westar plants were transformed with 3-oxoacyl-ACP reductase (KR) in antisense orientation, driven by either the cauliflower mosaic virus 35S promoter or a seed-specific acyl carrier protein promoter to determine the effects on plant productivity and on the activity of other fatty acid synthase (FAS) components. In plants with altered KR activity, total seed yield was reduced in all cases. In less severely affected plant lines, seeds had a normal appearance and composition but the yield of seeds was reduced by approximately 50%. In more severely affected lines, reductions in both seed fatty acid content and the number of seeds produced per plant were evident, resulting in a 90% reduction in fatty acid synthesized per plant. These phenotypes were independent of the promoter used. In severely affected lines, a large proportion of seeds showed precocious germination, and these had a reduced oleate content and increased levels of polyunsaturated 18-carbon fatty acids, compared with normal seeds of the same line. This reduction in 18:1 fatty acids was mimicked on imbibition of seeds with a normal appearance, indicating a preferential use of oleate moieties in precocious germination events. The reduction in activity of KR was mirrored for a second fatty acid synthase component, enoyl-ACP reductase, indicating a mechanism to maintain the ratio of fatty acid synthase components throughout embryogenesis. PMID- 17401136 TI - The role and aims of the FYSSION project. AB - FYSSION is a resource for researchers working on the fission yeast Schizosaccharomyces pombe. It currently comprises libraries of temperature sensitive mutants in essential genes, and insertional mutants in non-essential genes, available for screening by visiting workers. Here we outline methods for constructing and using the libraries, and describe future prospects for functional genomics of this organism, here and elsewhere. PMID- 17401137 TI - Prostaglandin E2-EP4 receptor promotes endothelial cell migration via ERK activation and angiogenesis in vivo. AB - Prostaglandin E2 (PGE(2)), a major product of cyclooxygenase, exerts its functions by binding to four G protein-coupled receptors (EP1-4) and has been implicated in modulating angiogenesis. The present study examined the role of the EP4 receptor in regulating endothelial cell proliferation, migration, and tubulogenesis. Primary pulmonary microvascular endothelial cells were isolated from EP4(flox/flox) mice and were rendered null for the EP4 receptor with adenoCre virus. Whereas treatment with PGE(2) or the EP4 selective agonists PGE(1)-OH and ONO-AE1-329 induced migration, tubulogenesis, ERK activation and cAMP production in control adenovirus-transduced endothelial EP4(flox/flox) cells, no effects were seen in adenoCre-transduced EP4(flox/flox) cells. The EP4 agonist-induced endothelial cell migration was inhibited by ERK, but not PKA inhibitors, defining a functional link between PGE(2)-induced endothelial cell migration and EP4-mediated ERK signaling. Finally, PGE(2), as well as PGE(1)-OH and ONO-AE1-329, also promoted angiogenesis in an in vivo sponge assay providing evidence that the EP4 receptor mediates de novo vascularization in vivo. PMID- 17401138 TI - Nonexercise movement in elderly compared with young people. AB - The association between free-living daily activity and aging is unclear because nonexercise movement and its energetic equivalent, nonexercise activity thermogenesis, have not been exhaustively studied in the elderly. We wanted to address the hypothesis that free-living nonexercise movement is lower in older individuals compared with younger controls matched for lean body mass. Ten lean, healthy, sedentary elderly and 10 young subjects matched for lean body mass underwent measurements of nonexercise movement and body posture over 10 days using sensitive, validated technology. In addition, energy expenditure was assessed using doubly labeled water and indirect calorimetry. Total nonexercise movement (acceleration arbitrary units), standing time, and standing acceleration were significantly lower in the elderly subjects; this was specifically because the elderly walked less distance per day despite having a similar number of walking bouts per day compared with the young individuals. The energetic cost of basal metabolic rate, thermic effect of food, total daily energy expenditure, and nonexercise activity thermogenesis were not different between the elderly and young groups. Thus, the energetic cost of walking in the elderly may be greater than in the young. Lean, healthy elderly individuals may have a biological drive to be less active than the young. PMID- 17401139 TI - Reply to letter "Pyruvate metabolism in working human skeletal muscle" by Henderson et al. PMID- 17401141 TI - Keratinocyte-specific expression of fatty acid transport protein 4 rescues the wrinkle-free phenotype in Slc27a4/Fatp4 mutant mice. AB - FATP4 (fatty acid transport protein 4; also known as SLC27A4) is the most widely expressed member of a family of six long chain fatty acid transporters. FATP4 is highly expressed in enterocytes and has therefore been proposed to be a major importer of dietary fatty acids. Two independent mutations in Fatp4 cause mice to be born with thick, tight, shiny, "wrinkle-free" skin and a defective skin barrier; they die within hours of birth from dehydration and restricted movements. In contrast, induced keratinocyte-specific deficiency of FATP4 in adult mice causes only mild skin abnormalities. Therefore, whether the loss of FATP4 from skin or a systemic gestational metabolic defect causes the severe skin defects and neonatal lethality remain important unanswered questions. To investigate the basis for the phenotype, we first generated wild-type tetraploid/mutant diploid aggregates that should lead to rescue of any abnormalities caused by loss of FATP4 from the placenta. However, the skin phenotype was not ameliorated. We then generated transgenic mice expressing exogenous FATP4 either widely or specifically in suprabasal keratinocytes, and we bred the transgenes onto the Fatp4(-/-) background. Both modes of FATP4 expression led to rescue of the neonatally lethal skin defects, and the resulting mice were viable and fertile. Keratinocyte expression of an FATP4 variant with mutations in the acyl-CoA synthetase domain did not provide any degree of rescue. We conclude that expression of FATP4 with an intact acyl-CoA synthetase domain in suprabasal keratinocytes is necessary for normal skin development and that FATP4 functions in establishing the cornified envelope. PMID- 17401142 TI - The C terminus of apolipoprotein A-V modulates lipid-binding activity. AB - Human apolipoprotein A-V (apoA-V) is a potent modulator of plasma triacylglycerol (TG) levels. To probe different regions of this 343-amino-acid protein, four single Trp apoA-V variants were prepared. The variant with a Trp at position 325, distal to the tetraproline sequence at residues 293-296, displayed an 11-nm blue shift in wavelength of maximum fluorescence emission upon lipid association. To evaluate the structural and functional role of this C-terminal segment, a truncated apoA-V comprising amino acids 1-292 was generated. Far UV circular dichroism spectra of full-length apoA-V and apoA-V-(1-292) were similar, with approximately 50% alpha-helix content. In guanidine HCl denaturation experiments, both full-length and truncated apoA-V yielded biphasic profiles consistent with the presence of two structural domains. The denaturation profile of the lower stability component (but not the higher stability component) was affected by truncation. Truncated apoA-V displayed an attenuated ability to solubilize l alpha-dimyristoylphosphatidylcholine phospholipid vesicles compared with full length apoA-V, whereas a peptide corresponding to the deleted C-terminal segment displayed markedly enhanced kinetics. The data support the concept that the C terminal region is not required for apoA-V to adopt a folded protein structure, yet functions to modulate apoA-V lipid-binding activity; therefore, this concept may be relevant to the mechanism whereby apoA-V influences plasma TG levels. PMID- 17401143 TI - Risk of cardiac catheterization under anaesthesia in children with pulmonary hypertension. AB - BACKGROUND: Children with primary pulmonary hypertension (PHT) are a high-risk group who require assessment by cardiac catheterization under anaesthesia. Complications, including death, have occurred during anaesthesia in these patients, but the true risk has not been quantified. METHODS: The clinical records of children with PHT undergoing general anaesthesia for pulmonary vascular resistance studies were reviewed retrospectively. Data collected included pre-catheter measures of severity of disease, details of clinical management, and complications occurring within 24 h of the start of anaesthesia. RESULTS: During the past 5 yr, 75 consecutive patients were catheterized and usable records were available in 70. The age range was 0.1-18 yr (mean 7.1). Four children required external cardiac massage [6% (95% confident limits 1-11%)] and one of these died. Of the four, two had an arrhythmia related to the mechanical effects of catheterization, one was hypotensive during anaesthesia and the other had fatal cardiac failure in recovery. All four had severe PHT as judged by echocardiographic estimation of tricuspid regurgitant jet velocity>4 m s-1. CONCLUSIONS: Resuscitation or death occurred in 6% of cases. Any associated risk factors could not be determined because the number of complications was too small. Risks may be highest in children with severe idiopathic PHT and symptoms of chest pain, syncope, or dizziness. PMID- 17401144 TI - Ubiquinone-10 production using Agrobacterium tumefaciens dps gene in Escherichia coli by coexpression system. AB - Ubiquinone (Coenzyme Q; abbreviation, UQ) acts as a mobile component of the respiratory chain by playing an essential role in the electron transport system, and has been widely used in pharmaceuticals. The biosynthesis of UQ involves 10 sequential reactions brought about by various enzymes. In this study we have cloned, expressed the decaprenyl diphosphate synthase, designated dps gene, from Agrobacterium tumefaciens, and succeeded in detecting UQ-10 in addition to innate UQ-8 in Escherichia coli. Furthermore, the production of UQ-10 was higher than UQ 8. To establish an efficient expression system for UQ- 10 production, we used genes, including ubiC, ubiA, and ubiG involved in UQ biosynthesis in E. coli, to construct a better co-expression system. The expression coupled by dps and ubiCA was effective for increasing UQ-10 production by five times than that by expressing single dps gene in the shake flask culture. To study for a large-scale production of UQ-10 in E. coli, fed-batch fermentations were implemented to achieve a high cell density culture. A cell concentration of 85.40 g/L and 94.58 g/L dry cell weight (DCW), and UQ-10 content of 50.29 mg/L and 45.86 mg/L was obtained after 32.5 h and 27.5 h of cultivation, subsequent to isopropyl-beta-D thiogalactopyranoside and lactose induction, respectively. In addition, plasmid stability was maintained at high level throughout the fermentation. PMID- 17401145 TI - Generation of linear expression constructs by one-step PCR with vaccinia DNA topoisomerase I. AB - Linear expression constructs can facilitate gene function studies. We describe a method to generate linear expression constructs for mammalian cells by one-step polymerase chain reaction (PCR) with vaccinia DNA topoisomerase I (TOPO). Cytomegalovirus (CMV) 5?' promoter, the gene of interest, and V5 bovine growth hormone (BGH) polyA 3?' terminator elements were PCR-amplified with target specific primers containing vaccinia DNA TOPO-specific sequence and complementary sequence to each other. We amplified specific and complementary sequences. These three elements were directionally joined with vaccinia TOPO. The joined products were then directly transfected into Chinese hamster ovary cells. Compared with the transfection of supercoiled plasmids, comparable expression signals were obtained for green fluorescent protein, chloramphenicol acetyltransferase, and beta-galactosidase proteins using Western blots. This is a quick and efficient method to generate linear expression constructs. Unlike Invitrogen TOPO Tools, our method avoided the secondary round of PCR and more rapidly yielded correct joining products. This method can be easily used in the function test of uncharacterized open reading frames. PMID- 17401146 TI - Expression in Pichia pastoris and immunological evaluation of a truncated Dengue envelope protein. AB - Among the Dengue virus structural proteins, the Envelope glycoprotein is the most important because of its antigenic characteristics. In this work, the E protein from Dengue-2 virus truncated at the C-terminus region was successfully expressed in Pichia pastoris. The E2trunc gene was cloned under the AOX1 promoter from P. pastoris and the signal peptide of the sucrose invertase gene from Saccharomyces cerevisiae. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of expression revealed the presence of a protein with the expected size, which was completely associated to the insoluble fraction after cellular disruption. The recombinant N-glycosylated protein reacted with two conformational antibodies against Dengue-2, indicating a proper folding of it. In addition, it was able to induce antiviral antibodies after mice immunization. PMID- 17401147 TI - Mitochondrial DNA variability and development of a PCR diagnostic test for populations of the whitefly Bemisia afer (Priesner and Hosny). AB - The whitefly, Bemisia afer (Hemiptera; Aleyrodidae), is emerging as a major agricultural pest. The current identification methods based on adult and pupal morphology are laborious and unreliable. A diagnostic polymerase chain reaction (PCR) protocol was developed for the first time in this study to discriminate B. afer from other whitefly species. Primers specific to mitochondrial cytochrome oxidase I gene (mtCOI) were designed to amplify a band of approx 650 bp. The PCR products were sequenced from B. afer samples collected from Malawi, Tanzania, Uganda, Zanzibar, and the United Kingdom. Phylogenetic analyses of mtCOI sequences and those of reference B. afer sequences clustered the African B. afer separately from the UK and Chinese populations and from other whitefly species. The African cluster was divided into two clades by parsimony and neighbor-joining methods. This indicates the existence of at least two genotypic clusters of B. afer, which are diverged by 0.8 to 3.2% nucleotide (nt) identities. Analysis of molecular variance indicated that these differences were the result of within population variation but were insufficient to identify discrete populations. Among the whitefly species used in the analysis, B. afer was equally dissimilar to Bemisia tabaci and Bemisia tuberculata (21.3-26.2% nt identities). As is the case for B. tabaci, these data show that mtCOI sequences are informative also for identifying B. afer variants, which lack distinguishing morphological features. PMID- 17401148 TI - High level expression of recombinant Mycobacterium tuberculosis culture filtrate protein CFP32 in Pichia pastoris. AB - Difficulty in obtaining large quantities of Mycobacterium tuberculosis (MTB) proteins remains a major obstacle in the development of subunit vaccines and diagnostic reagents for tuberculosis. A major reason is because Escherichia coli has not proven to be an optimal host for the expression of MTB genes. In this article, we used the yeast Pichia pastoris to express high levels of CFP32, a culture filtrate protein restricted to the MTB complex and a potential target antigen for serodiagnosis of tuberculosis in patients. Using shaker flasks, we generated a P. pastoris clone expressing CFP32 as a secreted protein fused to the myc- (His)6 tag, at a yield of 0.5 g of purified protein per liter of culture. Recombinant CFP32 (rCFP32) produced in P. pastoris has a molecular weight of 35 kDa, which is slightly higher than that of the native protein. We identified putative acylation and glycosylation sites in the CFP32 amino acid sequence that suggested posttranslational modifications may contribute to the size difference. The NH2-terminal peptide sequencing of rCFP32 showed that the signal peptide alpha factor is correctly excised. In addition, rCFP32 reacted with the sera of patients with tuberculosis. These data are the first to show that P. pastoris is a suitable host for high-yield production of good quality mycobacterium antigens, and especially culture filtrate proteins that have vaccine and diagnostic potential. PMID- 17401149 TI - Novel short peptides isolated from phage display library inhibit vascular endothelial growth factor activity. AB - Signal transduction through the vascular endothelial growth factor (VEGF)-VEGF receptor (VEGFR) pathway has a pivotal importance in angiogenesis, and has therefore become a prime target in antitumor therapy. In search for peptides antagonizing VEGF binding to its receptors, we screened a random heptamer library displayed on phage for peptides that bind the whole VEGF165 molecule and inhibit VEGF dependent human umbilical vein endothelial cell (HUVEC) proliferation. Two selected peptides with sequences WHLPFKC and WHKPFRF were synthesized. Biacore and matrix-assisted laser desorption/ionization timeof- flight mass spectrometry analysis indicated that these peptides bind the VEGF homodimer in a concentration dependent manner, with micromolar affinity, and with a 2:1 peptide:VEGF stoichiometry. They inhibited HUVEC proliferation in vitro by 77 and 55%, respectively. Taken together, our results indicate that these peptides could be potent inhibitors of angiogenesis. Furthermore, we show that the peptide- VEGF binding properties can be quantified, a prerequisite for the further optimization of binders. PMID- 17401150 TI - Online resources for SNP analysis: a review and route map. AB - The major online single nucleotide polymorphism (SNP) databases freely available as research tools for genetic analysis are explained, reviewed, and compared. An outline is given of the search strategies that can be used with the most extensive current SNP databases: National Centre for Biotechnology Information (NCBI) dbSNP and HapMap to help the user secure the most appropriate data for the research needs of clinical genetics and population genetics research. A range of online tools that can be useful in designing SNP genotyping assays are also detailed. PMID- 17401152 TI - The insertion polymorphism in angiotensin-converting enzyme gene associated with the APOE epsilon 4 allele increases the risk of late-onset Alzheimer disease. AB - Several studies have shown that a common insertion (I)/deletion (D) polymorphism of angiotensin-converting enzyme (ACE) gene may confer an increased risk of late onset Alzheimer disease (LOAD). However, the result has not been replicated by all studies. In order to clarify the role of the polymorphism for the occurrence of LOAD in Chinese and the possibility of a synergistic effect with the apolipoprotein E allele 4 on the risk of Alzheimer disease, we examined the ACE and APOE genotypes in a Chinese sample consisting of 104 sporadic LOAD patients and 128 healthy controls. An obvious difference of allelic and genotypic distributions of ACE I/D polymorphism between cases and controls was observed (chi2 = 6.61, df = 2, p = 0.037 by genotype; chi2 = 4.67, df = 1, p = 0.031 by allele). And ACE I allele carriers showed an increased risk for LOAD developing (chi2 = 6.59, df = 1, p =0.01, OR = 2.91, 95% CI 1.25-6.77). After stratifying by APOE epsilon 4 status, the increased LOAD risks associated with I allele carriers only in the APOE epsilon 4 noncarriers was seen (chi2 = 4.12, df = 1, p = 0.042). Logistic regression analysis of total subjects demonstrated a more than sevenfold increase in the risk of developing LOAD in subjects carrying both the ACE I allele and the APOE epsilon 4 (OR = 7.39, 95% CI 2.50-21.89, p < 0.001). Our data revealed that ACE I/D polymorphism is considered to be an additional risk factor, which has strong synergistic interaction with APOE epsilon 4 on the risk of LOAD. PMID- 17401151 TI - Chaperones and proteases: cellular fold-controlling factors of proteins in neurodegenerative diseases and aging. AB - The formation of toxic protein aggregates is a common denominator to many neurodegenerative diseases and aging. Accumulation of toxic, possibly infectious protein aggregates induces a cascade of events, such as excessive inflammation, the production of reactive oxygen species, apoptosis and neuronal loss. A network of highly conserved molecular chaperones and of chaperone-related proteases controls the fold-quality of proteins in the cell. Most molecular chaperones can passively prevent protein aggregation by binding misfolding intermediates. Some molecular chaperones and chaperone-related proteases, such as the proteasome, can also hydrolyse ATP to forcefully convert stable harmful protein aggregates into harmless natively refoldable, or protease-degradable, polypeptides. Molecular chaperones and chaperone-related proteases thus control the delicate balance between natively folded functional proteins and aggregation-prone misfolded proteins, which may form during the lifetime and lead to cell death. Abundant data now point at the molecular chaperones and the proteases as major clearance mechanisms to remove toxic protein aggregates from cells, delaying the onset and the outcome of protein-misfolding diseases. Therapeutic approaches include treatments and drugs that can specifically induce and sustain a strong chaperone and protease activity in cells and tissues prone to toxic protein aggregations. PMID- 17401153 TI - Characterization of 5-HT3 receptor mutations identified in schizophrenic patients. AB - The 5-HT3 receptor is a member of the Cys-loop family of ligand-gated ion channels, proteins that have been implicated in the pathology of several neurological disorders. In this study, we examine two mutations (R344H and P391R) that have been previously identified in individuals diagnosed with schizophrenia. These mutations are located in the M3-M4 loop of the 5-HT3 receptor and their occurrence presents the possibility that they contribute toward the etiology of this disorder. Radioligand binding with the 5-HT receptor antagonist, [3H]granisetron, revealed no significant difference in receptor affinity or density between mutant and wild-type receptors when expressed in HEK293 cells. However, comparison of EC50 values using whole-cell patch clamp for wild-type (1.68 microM +/- 0.01, n = 38), R344H (1.70 microM +/- 0.02, n = 18), and P391R (2.73 microM +/- 0.01, n = 8) receptors revealed a significant increase in the EC50 of the P391R mutant. Analysis of Hill co-efficients, and activation and desensitization rate constants showed no significant difference between wild-type and mutant receptors. These data suggest that the P391R, but not the R344H, mutation may play a role in the pathology of schizophrenia. PMID- 17401154 TI - Sex differences in lumbar spinal cord gene expression following experimental lumbar radiculopathy. AB - Considerable evidence indicates that there are sex-related differences in clinical and experimental pain sensitivity. In the present study, we sought to determine what genes were expressed in the spinal cord in a sexually dimorphic manner. We first analyzed global gene expression in the lumbar spinal cord of uninjured male and female rats using the Affymetrix RAE230A GeneChip platform in order to identify genes that are selectively expressed in male and female rats at a basal level. We subsequently analyzed global gene expression in the lumbar spinal cord of male and female rats at two time points (7 days and 14 d) following a rodent model of lumbar radiculopathy (L5 nerve root ligation) in order to determine what genes were regulated in a sexually dimorphic manner following nerve root injury. We utilized a linear regression analysis method to identify genes that were significantly different from the corresponding sham surgical controls. The expression patterns of several genes of interest were subsequently confirmed using RT-PCR. Our findings demonstrate significant differences in lumbar spinal cord gene expression in both uninjured and injured (L5 nerve root ligation) male and female rats. Further confirmation of a subset of the genes identified Neuregulin 1 and its high affinity receptor, ErbB4, Tachykinin 1, and Metabotropic glutamate receptor 6 as female specific genes upregulated following L5 nerve root injury. These findings provide several target genes for further study that may elucidate the neurochemical mechanisms underlying sex differences in pain sensitivity and lead to improved treatments for chronic pain syndromes. PMID- 17401155 TI - Oxidative stress and 17-alpha- and 17-beta-estradiol modulate neurofilaments differently. AB - Oxidative stress plays an important role in the pathogenesis of neurodegenerative diseases such as Parkinson's disease (PD). Neuronal death in the substantia nigra of PD patients is partly caused by exacerbated oxidative damage. Our previous studies demonstrated that oxidative stress can alter the structure and stability of neurofilament (NF) proteins and that 17-alpha- and 17-beta-estradiol are potent neuroprotective agents. The aim of this study was to investigate the cytoskeletal target of neuroprotection by estrogens in neuronal PC12 cells. We induced oxidative stress by MPP+ administration for 24 h, and 17-alpha- and 17 beta-estradiol were used as neuroprotective drugs. We measured gene expression and protein expression of each NF subunit, NFL, NFM, and NFH, by semiquantitative RT-PCR, Western blot, and immunofluorescence. Our results demonstrate that NFL mRNA and protein levels are not modulated by MPP+ or estradiol isomers, whereas NFM gene expression, as well as protein expression, are strongly influenced by MPP+, 17-alpha-, and 17-beta-estradiol after a 24-h treatment. Finally, mRNA levels of the most phosphorylated subunits, NFH, are not changed by MPP+ or treatment with both estradiol isomers, whereas NFH protein expression is decreased by the same treatments. These results suggest that oxidative stress affects neuronal cytoskeleton, maybe though proteolysis and/or abnormal structural changes in NFs. Then, 17-alpha- and 17-beta-estradiol might help the neuronal cell in recovering after oxidative stress by inducing protein expression of NFM and NFH subunits. PMID- 17401156 TI - Changes in cholesterol metabolism are associated with PS1 and PS2 gene regulation in SK-N-BE. AB - Several lines of evidence suggest that the cholesterol content of neuronal membranes influences amyloid precursor protein (APP) processing; however, its role in transcriptional regulation of the cofactors for gamma-secretase, the key enzyme for the production of the Abeta peptide, is poorly understood. This study investigates whether the changes in cellular cholesterol metabolism modulate the expression of genes involved in the gamma-secretase complex function. The abundance of mRNA transcripts for presenilin 1 and 2 (PS1 and PS2), APP, and nicastrin were evaluated in neuroblastoma cells exposed either to serum-depleted medium or to low-density lipoproteins (LDL). Cholesterol esterification was markedly inhibited by mevinolin and U18666A, but was not significantly affected by any other of the tested treatments. gamma-Secretase genes and cofactors were not co-regulated and were not influenced by statin inhibition of cholesterol synthesis. Nicastrin and the APP isoforms showed constitutive expression. In the absence of exogenous lipids, cell PS1 and PS2 expression was induced by LDL and by lysosomal sequestration of cholesterol. However, a different pattern of induction of presenilin gene expression was observed in the latter condition, suggesting that lysosomal cholesterol levels are strong inducers of PS2 transcription. Taken together, these results indicate that lipid metabolism has a complex influence on gamma-secretase transcriptional pathways and, in particular, exogenous cholesterol and compartmentalization in neuroblastoma cells play a relevant role in regulating the transcription of presenilins, while modulation of the cholesterol biosynthesis pathway seems to exert a minor influence on the expression of gamma-secretase genes and cofactors. PMID- 17401157 TI - Association of the 5-HT2A receptor gene polymorphism 102T/C with ischemic stroke. AB - Serotonin (5-HT) has been implicated in a number of cardiovascular disorders due to its ability to induce vascular contraction and platelet aggregation through activation of the 5-HT2 receptor family. In this study, we investigated the association of stroke in a Scandinavian population with two common polymorphisms in the 5-HT2A receptor gene. The two polymorphisms under investigation, namely the 102T/C and the -1438A/G variations of the 5-HT2A receptor gene, were examined in a case control association study involving 99 stroke patients and a comparable number of controls. Among patients, the prevalence of the homozygous 102T/T genotype was significantly higher than in controls (28.3% vs 13.5%; p < 0.01). The allelic frequency of 102T carriers was also significantly higher in stroke patients than in controls (p = 0.002, OR = 1.88, 95% CI, 1.27-2.80). The association between the 102T allele and stroke was significant in both males and females. There was no association between stroke and the -1438A/G polymorphism. Taken together, this study indicates that the 102T/C polymorphism in the 5-HT2A receptor gene could be an independent risk factor for developing stroke. PMID- 17401158 TI - Neonatal mice of the Down syndrome model, Ts65Dn, exhibit upregulated VIP measures and reduced responsiveness of cortical astrocytes to VIP stimulation. AB - The Ts65Dn segmental mouse model of Down syndrome (DS) possesses a triplication of the section of chromosome 16 that is most homologous to the human chromosome 21 that is trisomic in DS. This model exhibits many of the characteristics of DS including small size, developmental delays, and a decline of cholinergic systems and cognitive function with age. Recent studies have shown that vasoactive intestinal peptide (VIP) systems are upregulated in aged Ts65Dn mice and that VIP dysregulation during embryogenesis is followed by the hypotonia and developmental delays as seen in both DS and in Ts65Dn mice. Additionally, astrocytes from aged Ts65Dn brains do not respond to VIP stimulation to release survival-promoting substances. To determine if VIP dysregulation is age-related in Ts65Dn mice, the current study examined VIP and VIP receptors (VPAC-1 and VPAC-2) in postnatal day 8 Ts65Dn mice. VIP and VPAC-1 expression was significantly increased in the brains of trisomic mice compared with wild-type mice. VIP-binding sites were also significantly increased in several brain areas of young Ts65Dn mice, especially in the cortex, caudate/putamen, and hippocampus. Further, in vitro treatment of normal neurons with conditioned medium from VIP-stimulated Ts65Dn astrocytes from neonatal mice did not enhance neuronal survival. This study indicates that VIP anomalies are present in neonatal Ts65Dn mice, a defect occurs in the signal transduction mechanism of the VPAC-1 VIP receptor, cortical astrocytes from neonatal brains are dysfunctional, and further, that VIP dysregulation may play a significant role in DS. PMID- 17401159 TI - Effects of prolonged treatment with the opiate tramadol on prodynorphin gene expression in rat CNS. AB - A low abuse liability is reported for tramadol, an analgesic drug centrally acting through either opioid or nonopioid mechanisms. In this paper, we evaluated the effects of the repeated administration (7 d) of different doses of tramadol (10, 20, and 80 mg/kg, intraperitoneally) on the opioid precursor prodynorphin biosynthesis, in comparison with morphine (10 mg/kg, intraperitoneally), in the rat central nervous system (CNS). Northern analysis showed that morphine and tramadol produced different effects. While morphine caused a downregulation of prodynorphin mRNA levels in all investigated areas (hypothalamus, hippocampus, and striatum), tramadol did not cause any significant change in the striatum, and did not decrease prodynorphin biosynthesis in the hypothalamus and in the hippocampus, at nontoxic doses (10 and 20 mg/kg). The highest dose of tramadol (80 mg/kg) decreased prodynorphin mRNA levels in the hypothalamus and the hippocampus but not in the striatum. These data give some information on tramadol effects at molecular level in the CNS. They indicate that the alterations of prodynorphin gene expression caused by tramadol and morphine show a different pattern that may be related to the different abuse potential of the two analgesic drugs. PMID- 17401160 TI - Manipulation of early region 4. AB - Adenovirus early region 4 (E4) regulates processes in infected cells that include viral late gene expression, nonhomologous end joining, responses to DNA damage, and apoptosis. E4 is essential for viral growth in most cell lines. In this chapter, the current knowledge of the functions of six E4 products is summarized briefly. Protocols are presented for manipulation of E4, incorporation of E4 mutations into the viral genome, and growth of E4 mutants on complementing cell lines. A compilation of the described E4-complementing cell lines is included. PMID- 17401161 TI - Isolation, growth, and purification of defective adenovirus deletion mutants. AB - Defective adenovirus deletion mutants can be grown by complementation in the presence of helper viruses that supply essential functions missing in the deletion mutant. In general, the deletion mutant then must be separated physically from the helper for use in subsequent experiments. This chapter includes suggestions for selection of helper viruses, protocols for the production of stocks by complementation, and procedures for physical separation of deletion mutants from their helpers. PMID- 17401162 TI - Construction of adenovirus type 5 early region 1 and 4 virus mutants. AB - This chapter describes a novel strategy that simplifies the generation and production of adenovirus type 5 (Ad5) mutants carrying defined mutations in early transcription units 1 (E1) and 4 (E4). The strategy involves three recombinant plasmids containing E1 (pE1-1235), E4 (pE4-1155), or the wild-type genome that lacks a portion of E3 (pH5pg4100). To generate recombinant viruses, mutations are first introduced into pE1- and/or pE4-transfer plasmids by site-directed mutagenesis. The mutagenized constructs are then ligated into plasmid pH5pg4100 containing the Ad backbone by direct cloning. Infectious viral DNAs are released from the recombinant plasmids by PacI-digestion and transfected into the complementing cell lines 293 or W162, and viral progeny are isolated and amplified. The advantages of this strategy are multiple: all cloning steps are carried out in Escherichia coli, and any genetic region of the viral E1 and/or E4 transcription units can be specifically modified or deleted. Moreover, foreign genes can be introduced into the E1 and/or E4 regions, and expression of viral or therapeutic genes can be controlled by cell-type specific and/or inducible promoters. PMID- 17401163 TI - Construction of mouse adenovirus type 1 mutants. AB - Mouse adenovirus provides a model for studying adenovirus pathogenesis in the natural host. The ability to make viral mutants allows the investigation of specific mouse adenoviral gene contributions to virus-host interactions. Methods for propagation and titration of wild-type mouse adenovirus, production of viral DNA and viral DNA-protein complex, and transfection of mouse cells to obtain mouse adenovirus mutants are described in this chapter. Plaque purification, propagation, and titration of the mutant viruses are also presented. PMID- 17401164 TI - Generation of recombinant adenovirus using the Escherichia coli BJ5183 recombination system. AB - One of the most time-consuming steps in the generation of adenoviral vectors is the construction of recombinant plasmids. This chapter describes a detailed method for the rapid construction of adenoviral vectors. The method described here uses homologous recombination machinery of Escherichia coli BJ5183 to construct plasmids used in generation of adenoviral vectors. With this method, no ligation steps are involved in generating the plasmids, and any region of the adenoviral genome can be easily modified. Briefly, the full-length adenoviral genome flanked by unique restriction enzyme sites is first cloned into a bacterial plasmid. Next, the region of the viral genome to be modified is subcloned into a bacterial shuttle plasmid, and the desired changes are introduced by molecular biology techniques. The modified viral DNA fragment is gel-purified and cotransformed with the full-length plasmid, linearized in the targeted region, into BJ5183 cells. Homologous recombination in E. coli generates plasmids containing the modified adenoviral genome. Recombinant virus is generated following release of the viral DNA sequences from the plasmid backbone and transfection into a producer cell line. With this method, homogeneous recombinant adenoviruses can be obtained without plaque purification. PMID- 17401165 TI - Production and release testing of ovine atadenovirus vectors. AB - Gene-directed enzyme prodrug therapy (GDEPT) is an emerging approach for the treatment of cancers. A variety of viral vectors have been used to deliver genes that encode the relevant enzymes, and some have been tested in clinical trials. To ensure the potency and efficacy of such vectors and to obtain regulatory approval to administer them to humans, it is necessary to develop a suite of assays that provide quality assurance. New GDEPT vectors based on ovine atadenovirus and Escherichia coli purine nucleoside phosphorylase (PNP) have been developed for first time use in humans in a phase I trial for the treatment of prostate cancer. Here we describe methods for their production together with several quality-control assays. In particular, a functional cell killing assay was devised to measure the potency of PNP-GDEPT vectors, the principles of which could easily be adapted to other systems. PMID- 17401166 TI - Construction of capsid-modified recombinant bovine adenovirus type 3. AB - Adenoviruses have become a popular vehicle for gene transfer into animal and human cells. However, wide prevalence of preexisting immunity to human adenovirus (HAdV) and the promiscuous nature of the virus have made the use of nonhuman adenoviruses an attractive alternative. Moreover, readministration of viral vectors is often required to maintain therapeutic levels of transgene expression, resulting in vector-specific immune responses. Although a number of features of bovine adenovirus (BAdV)-3 make it attractive for use as a vector in human vaccination, BAdV-3 transduces nonbovine cells, including human cells, poorly. However, genetic modification of capsid proteins (e.g., fiber, pIX) has helped in increasing the utility of BAdV-3 as a vector for transducing nonbovine cells. Here, we will describe the methods used to construct recombinant BAdV-3 expressing chimeric fiber or chimeric pIX proteins. PMID- 17401167 TI - Adenovirus capsid chimeras: fiber terminal exon insertions/gene replacements in the major late transcription unit. AB - The adenovirus major late transcription unit (MLTU) encodes the main structural capsid proteins. Expression from the MLTU is accomplished through alternative mRNA processing and use of a terminal exon coding strategy. The capsid proteins hexon, penton, and fiber contribute to efficient infection by adenovirus, and each contributes in some manner to the antiviral immune response against adenovirus infection. The ability to manipulate these genes affords one the opportunity to "detarget" adenovirus, to retarget adenovirus, and to alter immune recognition. In this chapter, we are presenting a terminal exon-replacement strategy that can be used to genetically manipulate capsid proteins expressed from the MLTU. An emphasis will be placed on manipulations of fiber as an intact terminal exon. PMID- 17401168 TI - Temperature-sensitive replication-competent adenovirus shRNA vectors to study cellular genes in virus-induced apoptosis. AB - The use of shRNA for knockdown of gene expression is a powerful method. In addition to transient transfection of RNA oligonucleotides, various DNA-based vectors that express short hairpin RNAs have been successfully used for efficient depletion of gene products. Replication-defective retrovirus and adenovirus (Ad) vectors have also gained wide usage. The extension of shRNA technology to replication-competent Ad would be desirable to investigate the role of various cellular genes in Ad replication. This approach is hampered because the effect of shRNA is neutralized by the Ad VA-RNA that is expressed at late stages after infection, and the infected cells are killed prior to significant depletion of some long-lived target gene products. We have constructed replication-competent Ad vectors for the depletion of the pro-apoptotic proteins BAX and BAK. We have modified a replication-defective Ad multivalent shRNA expression vector developed by Welgen, Inc. In our vector design, the multivalent shRNA expression cassette is contained in the E1B region. Additionally, we have incorporated a temperature sensitive mutation in the viral DBP gene (ts125). The use of this vector has resulted in efficient depletion of critical cellular apoptotic modulators, BAX and BAK. This vector may be useful to study the role of various cellular genes in Ad-induced apoptosis and viral replication. PMID- 17401169 TI - Evaluating apoptosis in tumor cells infected with adenovirus expressing p53 tumor suppressor gene. AB - This chapter describes several methods for recognizing apoptosis in tumor cells following infection with a replication-deficient adenovirus expressing the tumor suppressor gene p53. We include cytotoxicity assays and assays of apoptosis, including DNA-nucleosomal DNA fragmentation (DNA laddering), TUNEL, DAPI staining, analysis of the sub-G1 (subdiploid) population, and degradation of poly(ADP-ribose) polymerase (as assayed by Western blot). Although this is not a comprehensive list of protocols to evaluate apoptosis, we believe that these will cover the majority of conditions of apoptosis that may arise. The chapter also describes the characteristics of each technique, including the advantages and disadvantages of each method. PMID- 17401170 TI - Growth and purification of enteric adenovirus type 40. AB - The enteric adenoviruses of subgroup F (Ad40 and Ad41) pose some special problems of cultivation, as they cannot be readily passaged in many of the cell types used to propagate the more commonly used subgroup C serotypes (Ad2 and Ad5) and there is no standard plaque assay. Methods to propagate Ad40 in complementing cell lines and to evaluate infectivity and particle number are presented in this chapter. PMID- 17401171 TI - Immunocompetent, semi-permissive cotton rat tumor model for the evaluation of oncolytic adenoviruses. AB - Oncolytic adenovirus (Ad) vectors belong to a new class of cancer therapy agents that destroy cancer cells as part of the virus's lytic infectious cycle. In this chapter we describe an immunocompetent, semi-permissive cotton rat tumor model to evaluate the safety and efficacy of oncolytic Ad vectors. With this model one can investigate the effect of the host immune system on the vector-tumor interaction as well as the vector's effect on normal host cells in vivo. This chapter describes procedures for analyzing the growth and cytolytic properties of oncolytic Ad vectors in cotton rat cells in vitro. We discuss handling and husbandry issues and techniques for subcutaneous, intratumoral, and intravenous injection of cotton rats. We present methods for generating subcutaneous tumors in cotton rats and assessing the efficacy of Ad vectors upon intratumoral injection. Also, we discuss procedures for determining the biodistribution of a replicating Ad in cotton rats. PMID- 17401172 TI - Use of the Syrian hamster as an animal model for oncolytic adenovirus vectors. AB - Oncolytic adenoviruses (Ads) are promising candidates for cancer therapy. However, current animal models to evaluate these vectors have substantial limitations. Because Ad replication is generally species-specific, oncolytic Ads are usually examined in immunodeficient mice bearing human xenograft tumors. However, this model suffers because the animals are immunodeficient and normal and cancerous mouse tissues are poorly permissive to human Ad replication. We have recently developed a Syrian hamster model that is both immunocompetent and permissive to human Ad replication in normal and cancerous tissues. The Syrian hamster is also permissive for Ad5 replication in the lung, which is the natural site of infection in humans. Human Ads replicate well in vitro in the Syrian hamster cell lines examined and demonstrate significant antitumor efficacy following injection into Syrian hamster tumors in vivo. In this chapter we describe the maintenance of these Syrian hamster cell lines in culture and how to assess oncolytic Ad vector replication in these cells in vitro. We also describe detailed methods for growth of these cell lines as subcutaneous tumors, for intravenous and intratumoral injections in hamsters, and for evaluation of the efficacy, replication, and biodistribution of oncolytic Ad vectors following administration in hamsters. In addition, we describe how to assess replication in normal tissues such as the lungs and give helpful tips on handling, anesthesia, and general care of Syrian hamsters. PMID- 17401173 TI - A real-time PCR method to rapidly titer adenovirus stocks. AB - A critical step in working with adenovirus (Ad) and its vectors is the accurate, reproducible, sensitive, and rapid measurement of the amount of virus present in a stock. Titration methods fall into one of two categories: determination of either the infectious or the particle (infectious plus noninfectious) titer. Determining the infectious titer of a virus stock by plaque assay has important limitations, including cell line-, researcher-, and laboratory-dependent variation in titer, and the length of time required to perform the assay (2-4 wk). A major drawback of particle titration methods is the lack of consistent correlation between the resultant titer and the infectious titer. To overcome these problems, a rapid, sensitive, and reproducible real-time polymerase chain reaction (PCR) assay was developed that detects encapsidated full-length genomes. Importantly, there is a linear correlation between the titer determined by the realtime PCR assay and the infectious titer determined by a plaque assay. This chapter provides step-by-step guidance for preparing viral DNA, conducting the real-time PCR assay, and using the resultant data to calculate a viral titer. PMID- 17401174 TI - Detection and quantitation of subgroup C adenovirus DNA in human tissue samples by real-time PCR. AB - Advances in amplification techniques have revolutionized the ability to detect viruses both quantitatively and qualitatively and to study viral load. Real-time polymerase chain reaction (PCR) amplification depends on the ability to detect and quantify a fluorescent reporter molecule whose signal increases in proportion to the amount of amplification product generated. Recent advances have been made by using probes, such as TaqMan probes, to detect amplified products. Use of these probes offers confirmation of specificity of the PCR product. Here we describe a sensitive real-time PCR assay to quantify subgroup C adenoviral DNA in human lymphocytes derived from mucosal tissues removed in routine tonsillectomy or adenoidectomy. This chapter will describe in detail the methods used for these analyses. PMID- 17401175 TI - Flow cytometric detection of adenoviruses and intracellular adenovirus proteins. AB - Precise and simple assay of purified and crude preparations of human adenoviruses is essential for basic and gene therapy research. Previous bioassays used to quantitate adenoviruses (such as the plaque assay or fluorescent focus assay) are time-consuming and subjective in their interpretation. Here we describe a flow cytometric method that eliminates these disadvantages and provides a quantitative and reliable method of focus-forming unit (FFU) assay. PMID- 17401176 TI - Capture ELISA quantitation of mouse adenovirus type 1 in infected organs. AB - A capture enzyme-linked immunosorbent assay (ELISA) was optimized for identification of mouse adenovirus type 1 in infected brain homogenates. The ELISA method allows for a much faster quantitation of virus in infected organs than plaque assays. Methods for organ homogenization and the subsequent capture ELISA are described in this chapter. PMID- 17401177 TI - Preparation and titration of CsCl-banded adenovirus stocks. AB - Adenovirus research often requires purified high-titer virus stocks and accurate virus titers for use in experiments. Accurate titers are important for quantitative, interpretable, and reproducible results. This is especially true when there are comparisons of different mutant viruses following infection. This chapter details the large-scale preparation of adenovirus (either replication competent or replication-defective) in spinner cultures (e.g., KB, HeLa, or 293 cells). Protocols for harvesting cells and isolation of adenovirus by CsCl banding are presented. Methods for titering adenovirus by plaque assay are presented along with a discussion of how plaque assays can be used to determine the kinetics of cell killing and cytolysis by adenoviruses. PMID- 17401178 TI - [Morphology and spatial position of the mandible in complete unilateral cleft lip and palate individuals after one-stage repair of the cleft]. AB - AIM: To compare the size and spatial position of the mandible in unilateral cleft lip and palate (UCLP) individuals treated with one-stage repair of the lip and palate, with normal individuals. MATERIAL AND METHODS: 25 UCLP children aged 9-11 years, treated during the first year of life (average--9 months) compared with 25 subjects with normal morphology craniofacial skeleton. The measurements included: anterior and posterior facial height, mandibular plane angle, mandibular ramus, corpus and total length, Sella-Nasion-point B and Sella-Nasion-Pogonion angles. The difference between the groups was assessed by the T test for independent samples. RESULTS: The mandibular anterior surface was retruded (SNB and SNPog angles were approximately 3 degrees less than in controls). The other variables measured in the cleft group showed no difference in comparison to the controls. In the cleft group, standard deviation was higher for most measurements. CONCLUSIONS: Growth of the mandible in the UCLP subjects treated with a one-stage repair is close to normal. However, final verification of the mandibular growth will be possible after attainment of maturity. PMID- 17401179 TI - [Comparison of testicular volume before and after laparoscopic varicocelectomy in children and adolescents]. AB - Male infertility is the most frequent consequence of late diagnosis of varicocoele and not performing surgical varicocelectomy before puberty. The etiopathogenesis is still obscure. The anatomical conditions in men are conducive to the development of left varicocoele. The aim of this study is the assessment of the results of varicocelectomy on increase of testicular volume in adolescents with the grade II and III varicocoele and on improvement of potential fertility of young men. MATERIAL AND METHODS: In the period between 1996-2004, 62 children and adolescents with varicocoele and with left small testis underwent laparoscopic varicocelectomy. Unilateral varicocelectomy was performed in 51 (83.3%) while bilateral varicocectomy was performed in 11 (17.7%) patients. The average age was 15.6 years (13-17 years). Clinical diagnosis was based on: general physical examination, andrologic and sonographic (colour Doppler) examination where testis volume and clinical degree were assessed according to Dubin and Amelar score. Laparoscopic transperitoneal procedure was carried out using two surgical techniques: testicular vein and artery ligation or only testicular vein ligation. RESULTS: Left testis volume increase i.e. comparable volume of both testies was observed in 22 (75.9%) young men with grade II (p<0.001) and in 29 (87.9%) with grade III of clinical advancement of varicocoele (p<0.003). The increase of testis volume was noted in 43 (84.3%) patients: among 51 patients with unilateral varicocoele and in 8 (72.7%) among 11 boys operated with bilateral varicocele (p<0.001). The increase of testis volume was not significantly greater in boys who underwent spermatic artery sparing technique (p=0.85). Out of 62 patients, in 53 (85.5%) normozoospermy was observed (p<0.005). CONCLUSIONS: Laparoscopic ligation of testicular vein produced significant increase of hypotrophic testicular volume in 82.3% operated children. The presence of hypotrophic testies in adults, after varicocelectomy before puberty is accompanied by oligoasthenoteratozoospermy and high plasma FSH, LH levels and low plasma testosterone level. Assessment of the results of the varicocelectomy in children should include in the procreation period semen analysis and the estimation of plasma FSH, LH and T levels. PMID- 17401180 TI - [Is soluble thrombomodulin a molecular marker of endothelial cell injury in children and adolescents with arterial hypertension?]. AB - INTRODUCTION: Endothelial damage is an early step in the pathogenesis of atherosclerosis which begins in early childhood after exposure to atherogenic risk factors such as arterial hypertension. Atherosclerosis is becoming a common disease even in children and adolescents. Its progression may lead to very severe cardiovascular complications. Thrombomodulin (TM), a specific marker of endothelial cell damage, is a transmembranous glycoprotein with vasoprotective and anti-coagulant properties. TM-thrombin complex becomes an activator of protein C which inactivates factor Va and VIIIa and thereby inhibits the blood coagulation cascade. TM may be cleaved to its soluble (sTM) form by proinflammatory mediators and then detected in the circulation. AIM: The aim of the present study was to investigate if sTM plasma concentration--one of biochemical markers of endothelium injury--is higher in children and adolescents with arterial hypertension (AH). MATERIALS AND METHODS: We studied 32 children with hypertension (9 girls and 23 boys, age range 10.5-17.2 years), and 17 healthy controls, without family history of cardiovascular diseases. Hypertension was detected after 24 hrs ABPM (Ambulatory Blood Pressure Monitoring). We measured plasma concentration of sTM, blood lipids profile, body mass index (BMI) and, as sTM is excreted by the kidney, we also measured plasma level of creatinine and its clearance. RESULTS: Plasma concentration of sTM in the group with essential hypertension was significantly higher than that in the control group (4.01 +/-1.05 vs 3.42+/-0.4; p<0. 05). There were no significant associations between sTM and age or sex. Analyzing lipids profile (cholesterol, LDL-c, HDL-c, triglycerides) we did not find any differences in their levels between groups. We observed statistically significant correlation between sTM and systolic and diastolic blood pressure during the day and night. In addition BMI ratio was higher in AH group but there was no significant correlation between sTM and BMI. CONCLUSION: Statistically higher level of sTM in children with AH compared with healthy individuals makes us sure that endothelium cells, even in children who were shortly exposed to atherogenic risk factors such as essential hypertension, are noticeably damaged. These results constitute an additional signal that a lot of effort should be put into the endeavors to eliminate atherogenic risk factors in children in order to prevent cardiovascular events. PMID- 17401181 TI - [Age at menarche in girls with sense organs impairments]. AB - AIM: The aim of the study was to assess the age at menarche in girls with chronic diseases of senses organs. MATERIAL AND METHODS: The sample consisted of 179 girls (87 with hearing defects and 92 with sight defects) aged from 10 to 16 years of age. All subjects were pupils of Special Schools for Deaf Children and Youth or Centres of Training for Blind and Sight Impairment Children in Wroclaw and Krakow (Poland). Using the status quo method age at menarche was estimated. RESULTS: Median age at menarche via probit analysis was 12.62 +/- 0.18 for girls with hearing impairments and 13.11 +/- 0.21 for girls with impairments of sight. The earliest age at menarche was observed in deaf girls and blind girls (12.56 +/ 0.20 and 12.82 +/- 0.32, respectively). CONCLUSION: The more severe the impairment of the sense organ, the earlier was the age at menarche. PMID- 17401182 TI - [Physical growth of children and youth with hearing impairment]. AB - AIM: The aim of the study was to assess physical growth of children and youth with hearing impairments. MATERIAL AND METHODS: The material comprised anthropometric measurements (body height and body mass) and data from questionnaires collected from 186 children (87 girls and 99 boys) aged 10-16 years, with defective hearing. The subjects attended two Specialist Schools for Deaf Children and Youth in Wroclaw and Cracow (Poland). Based on extended medical questionnaires, information of hearing defects (hypoacusia or deafness) and their etiology (hereditary, congenital or post-natally acquired) were obtained. The measurements were standardized to the mean value and to the standard deviation of the control group of children from Warsaw. RESULTS: The standardized values of somatic traits of the subjects revealed deficient stature and body mass in children with defective hearing (Z=-0.54, Z=-0.18, respectively). The children with light and severe impairments of hearing manifested similar level of growth. CONCLUSIONS: Lower level of physical growth in children with hearing impairment is caused by the defect together with unfavourable socio-economic conditions of the subjects' families. The tendency of the highest body height deficiencies was observed to occur in the group of children with a hearing defect acquired after birth. PMID- 17401183 TI - [Neurofibromatosis--an inborn genetic disorder with susceptibility to neoplasia]. AB - Among different subtypes of neurofibromatosis (Nf), type 1 (Nf-1) predominates in frequency (approximately 97% of Nfs' patients) with an incidence of approximately 1 in 3500 live births. Nf-2, comprises 2% of the Nf population and is a very rare disease (1:40,000). Both are autosomal dominant disorders with 100% penetration, variable expression and 50% rate of new (de novo) mutations. The protein products of both, NF1 andNF2 genes are best known and the genes serve as tumour suppressors. Mutations result in a predisposition to develop a variety of tumours of the central and peripheral nervous systems, as well as other malignancies. Nf 2 is a multisystem genetic disorder associated with bilateral vestibular schwannomas, spinal cord schwannomas, meningiomas, gliomas, and juvenile cataracts with a paucity of cutaneous features, which are seen more consistently in Nf-1. In contrast to Nf-1, Nf-2 is associated with significant morbidity and decreased life span and a higher incidence of CNS tumours. However, morbidity and mortality rates in Nf-1 are not negligible. The cardinal features of Nf-1 are cafe-au-lait spots, axillary and inguinal freckling, cutaneous neurofibromas, and iris hamartomas (Lisch nodules). Optic gliomas and both malignant and benign peripheral nerve sheet tumours are the most common malignancies arising in Nf-1 patients. Among neurological symptoms epilepsy, intellectual disability and learning difficulty are also observed. Bone dysplasia results in scoliosis. There is no known medical treatment beneficial to both groups of patients. The mainstay of care for Nf patients is anticipatory guidance, and early detection and symptomatic treatment of disease complications. PMID- 17401184 TI - [Perception of self and parents of obese preschool children]. AB - THE AIM of the study is to present characteristics of the sense of identity and mental representation of parents of obese preschool children. MATERIAL AND METHODS: 21 children with obesity and 24 children with normal weight were investigated using the Children's Apperception Test--CAT-A. The CAT-A consists of 10 black and white pictures presenting animals in different situations, significant in the child's development and functioning. RESULTS: Significant differences were found in the sense of identity of the examined children in the area of sense of mental distinctness as well as socio-emotional and sexual aspects of the sense of identity. The mental representations of parents were characterized by differences in accessibility and sense of continuity as well as socio-emotional, sexual and activity-related aspects of mental representation. The differences were connected with sex and body mass of the examined children. CONCLUSIONS: Therapeutic indications for clinical practice with obese children have been formulated: motivating the family as a whole to partake in the therapeutic process, necessity of early intervention, different modes of diagnosis and therapy for boys and girls, prophylactic care for obese girls as a potential risk group for eating disorders. PMID- 17401185 TI - [Spontaneous splenic rupture in the course of infectious mononucleosis]. AB - Spontaneous splenic rupture (SSR) in the course of infectious mononucleosis (IM) is a rare but potentially fatal complication. Mortality rate is relatively high, therefore emergency splenectomy is a life-saving intervention. In case of undergoing urgent operation there is no possibility to initiate proper prophylaxis of overwhelming infection. The humoral and cellular immunologic response impairment is a reason for life-threatening complications of splenectomised person. Asplenic children should receive infection prophylaxis immediately post splenectomy. We report two cases of splenic rupture inpatients with IM. The prevention of infection was initiated in both children. The mainstays of prophylaxis are: immunization, chemoprophylaxis and education. Complex information concerning asplenia-related subjects should be provided for patients and their parents. PMID- 17401186 TI - [The diagnostic difficulties in children with allergic skin changes and parvovirus B19 infection. Own experiences]. AB - The authors describe two children with similar skin changes, which have been provoked by two different diseases-atopic dermatitis and parvovirosis B19. The authors underline the coexistence of the two diseases in these children, diagnostic difficulties and the need to apply therapeutic management for both. PMID- 17401187 TI - [Gynecomastia--a frequent clinical problem]. AB - Gynecomastia is a frequent clinical problem. About 50% of cases of gynecomastia are known as physiological gynecomastia, connected with the neonatal period, puberty or aging. However, gynecomastia can also be the first symptom of a serious disease. In this review we present current opinions on the etiology, pathogenesis, diagnostics and treatment of gynecomastia with special regard to the new therapeutic possibilities. PMID- 17401188 TI - Genital mycoplasmas--morbidity and a potential influence on human fertility. AB - Mycoplasmas comprise a big group of organisms consisting of one hundred eighty species which are found in nature as parasites of humans, other mammals, reptiles, fishes and plants, or living as commensals. The group commonly referred to as genital mycoplasmas comprise species most often found in the genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can cause many different infections. The species we are most interested in this work are called: Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma urealyticum. There is no doubt that they can cause non-gonococcal urethritis (NGU) in both men and women and bacterial vaginosis (BV), cervicitis, endometritis. Infection can spread to the upper port of female genital tract and it can lead to pelvic inflammatory disease, or if it happens during pregnancy--to chorioamnionitis and further pregnancy complications. Even though mycoplasmas have been known and described since 1898, the problem of their morbidity and the possible influence they have on human fertility is still not clear. Similar to research from 30 years ago, connecting Chlamydia trachomatis with infertility, new scientific work as well as the dynamic development of diagnostics procedure, especially more common use of PCR method, may be helpful in discovering the potential role genital mycoplasmas play in infertility. PMID- 17401189 TI - Structure of the apo form of the catabolite control protein A (CcpA) from Bacillus megaterium with a DNA-binding domain. AB - Crystal structure determination of catabolite control protein A (CcpA) at 2.6 A resolution reveals for the first time the structure of a full-length apo-form LacI-GalR family repressor protein. In the crystal structures of these transcription regulators, the three-helix bundle of the DNA-binding domain has only been observed in cognate DNA complexes; it has not been observed in other crystal structures owing to its mobility. In the crystal packing of apo-CcpA, the protein-protein contacts between the N-terminal three-helix bundle and the core domain consisted of interactions between the homodimers that were similar to those between the corepressor protein HPr and the CcpA N-subdomain in the ternary DNA complex. In contrast to the DNA complex, the apo-CcpA structure reveals large subdomain movements in the core, resulting in a complete loss of contacts between the N-subdomains of the homodimer. PMID- 17401190 TI - Structure of the buffalo secretory signalling glycoprotein at 2.8 A resolution. AB - The crystal structure of a 40 kDa signalling glycoprotein from buffalo (SPB-40) has been determined at 2.8 A resolution. SPB-40 acts as a protective signalling factor by binding to viable cells during the early phase of involution, during which extensive tissue remodelling occurs. It was isolated from the dry secretions of Murrah buffalo. It was purified and crystallized using the hanging drop vapour-diffusion method with 19% ethanol as the precipitant. The protein was also cloned and its complete nucleotide and amino-acid sequences were determined. When compared with the sequences of other members of the family, the sequence of SPB-40 revealed two very important mutations in the sugar-binding region, in which Tyr120 changed to Trp120 and Glu269 changed to Trp269. The structure showed a significant distortion in the shape of the sugar-binding groove. The water structure in the groove is also drastically altered. The folding of the protein chain in the flexible region comprising segments His188-His197, Phe202-Arg212 and Tyr244-Pro260 shows large variations when compared with other proteins of the family. PMID- 17401191 TI - Structure determination and analysis of a bacterial chymotrypsin from Cellulomonas bogoriensis. AB - The crystal structure of a secreted chymotrypsin from the alkaliphile Cellulomonas bogoriensis has been determined using data to 1.78 A resolution and refined to a crystallographic R factor of 0.167. The crystal structure reveals a large P1 substrate-specificity pocket, as expected for chymotrypsins. The structure is compared with close structural homologues. This comparison does not reveal clear reasons for the alkali tolerance of the enzyme, but the greater compactness of the structure and lowered hydrogen bonding may play a role. PMID- 17401192 TI - Complex structure of a bacterial class 2 histone deacetylase homologue with a trifluoromethylketone inhibitor. AB - Histone deacetylases (HDACs) have emerged as attractive targets in anticancer drug development. To date, a number of HDAC inhibitors have been developed and most of them are hydroxamic acid derivatives, typified by suberoylanilide hydroxamic acid (SAHA). Not surprisingly, structural information that can greatly enhance the design of novel HDAC inhibitors is so far only available for hydroxamic acids in complex with HDAC or HDAC-like enzymes. Here, the first structure of an enzyme complex with a nonhydroxamate HDAC inhibitor is presented. The structure of the trifluoromethyl ketone inhibitor 9,9,9-trifluoro-8-oxo-N phenylnonanamide in complex with bacterial FB188 HDAH (histone deacetylase-like amidohydrolase from Bordetella/Alcaligenes strain FB188) has been determined. HDAH reveals high sequential and functional homology to human class 2 HDACs and a high structural homology to human class 1 HDACs. Comparison with the structure of HDAH in complex with SAHA reveals that the two inhibitors superimpose well. However, significant differences in binding to the active site of HDAH were observed. In the presented structure the O atom of the trifluoromethyl ketone moiety is within binding distance of the Zn atom of the enzyme and the F atoms participate in interactions with the enzyme, thereby involving more amino acids in enzyme-inhibitor binding. PMID- 17401193 TI - Structure of Physarum polycephalum cytochrome b5 reductase at 1.56 A resolution. AB - Physarum polycephalum cytochrome b(5) reductase catalyzes the reduction of cytochrome b(5) by NADH. The structure of P. polycephalum cytochrome b(5) reductase was determined at a resolution of 1.56 A. The molecular structure was compared with that of human cytochrome b(5) reductase, which had previously been determined at 1.75 A resolution [Bando et al. (2004), Acta Cryst. D60, 1929 1934]. The high-resolution structure revealed conformational differences between the two enzymes in the adenosine moiety of the FAD, the lid region and the linker region. The structural properties of both proteins were inspected in terms of hydrogen bonding, ion pairs, accessible surface area and cavity volume. The differences in these structural properties between the two proteins were consistent with estimates of their thermostabilities obtained from differential scanning calorimetry data. PMID- 17401194 TI - Crystallization and preliminary X-ray crystallographic analysis of Escherichia coliglutaredoxin 2 in complex with glutathione and of a cysteine-less variant without glutathione. AB - Glutaredoxin 2 (Grx2) from Escherichia coli is larger in size than classical glutaredoxins. It is extremely efficient in the catalysis of reduced glutathione dependent disulfide reduction. A complex of Grx2 with reduced glutathione (GSH) has been crystallized. Data were collected to 1.60 A. The crystals belong to space group P3(2)21, with one Grx2-GSH complex in the asymmetric unit. The unit cell parameters are a = b = 50.10, c = 152.47 A. A Grx2 mutant, C9S/C12S, which cannot form a disulfide bond with GSH was also crystallized. The crystals diffracted to 2.40 A and belong to space group P2(1)2(1)2(1), with one molecule in the asymmetric unit. The unit-cell parameters are a = 28.16, b = 78.65, c = 89.16 A. PMID- 17401195 TI - Expression, purification, crystallization and preliminary X-ray diffraction analysis of the DDX3 RNA helicase domain. AB - DDX3 is a human RNA helicase that is involved in RNA processing and important human diseases. This enzyme belongs to the DEAD-box protein family, the members of which are characterized by the presence of nine conserved motifs including the Asp-Glu-Ala-Asp motif that defines the family. DDX3 has two distinct domains: an ATP-binding domain in the central region of the protein and a helicase domain in the carboxy-terminal region. The helicase domain of DDX3 was cloned and overexpressed in Escherichia coli. Crystallization experiments yielded crystals that were suitable for X-ray diffraction analysis. The final crystallization conditions were a reservoir solution consisting of 2 M ammonium sulfate, 0.1 M imidazole pH 6.4 plus 5 mM spermine tetrahydrochloride and a protein solution containing 10 mM HEPES, 500 mM ammonium sulfate pH 8.0. The crystals of the helicase domain belong to the monoclinic space group P2(1), with unit-cell parameters a = 43.85, b = 60.72, c = 88.39 A, alpha = gamma = 90, beta = 101.02 degrees , and contained three molecules per asymmetric unit. These crystals diffracted to a resolution limit of 2.2 A using synchrotron radiation at the European Synchrotron Radiation Facility (ESRF) and the Swiss Light Source (SLS). PMID- 17401196 TI - Crystallization and preliminary X-ray crystallographic analysis of the heterodimeric crotoxin complex and the isolated subunits crotapotin and phospholipase A2. AB - Crotoxin, a potent neurotoxin from the venom of the South American rattlesnake Crotalus durissus terrificus, exists as a heterodimer formed between a phospholipase A(2) and a catalytically inactive acidic phospholipase A(2) analogue (crotapotin). Large single crystals of the crotoxin complex and of the isolated subunits have been obtained. The crotoxin complex crystal belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 38.2, b = 68.7, c = 84.2 A, and diffracted to 1.75 A resolution. The crystal of the phospholipase A(2) domain belongs to the hexagonal space group P6(1)22 (or its enantiomorph P6(5)22), with unit-cell parameters a = b = 38.7, c = 286.7 A, and diffracted to 2.6 A resolution. The crotapotin crystal diffracted to 2.3 A resolution; however, the highly diffuse diffraction pattern did not permit unambiguous assignment of the unit-cell parameters. PMID- 17401197 TI - Expression, crystallization and preliminary crystallographic data analysis of filamin A repeats 14-16. AB - Human filamin A is a 280 kDa protein involved in actin-filament cross-linking. It is structurally divided into an actin-binding headpiece (ABD) and a rod domain containing 24 immunoglobulin-like (Ig) repeats. A fragment of human filamin A (Ig repeats 14-16) was cloned and expressed in Escherichia coli and the purified protein was crystallized in 1.6 M ammonium sulfate, 2% PEG 1000 and 100 mM HEPES pH 7.5. The crystals diffracted to 1.95 A and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 50.63, b = 52.10, c = 98.46 A, alpha = beta = gamma = 90 degrees. PMID- 17401198 TI - Crystallization and preliminary crystallographic analysis of a haloalkane dehalogenase, DbjA, from Bradyrhizobium japonicum USDA110. AB - Haloalkane dehalogenases are key enzymes for the degradation of halogenated aliphatic pollutants. The haloalkane dehalogenase DbjA constitutes a novel substrate-specificity class with high catalytic activity for beta-methylated haloalkanes. In order to reveal the mechanism of its substrate specificity, DbjA has been crystallized using the hanging-drop vapour-diffusion method. The best crystals were obtained using the microseeding technique with a reservoir solution consisting of 17-19.5%(w/v) PEG 4000, 0.2 M calcium acetate and 0.1 M Tris-HCl pH 7.7-8.0. The space group of the DbjA crystal is P2(1)2(1)2, with unit-cell parameters a = 212.9, b = 117.8, c = 55.8 A. The crystal diffracts to 1.75 A resolution. PMID- 17401199 TI - Preliminary structural studies on the leucine-zipper homology region of the human protein Bap31. AB - B-cell receptor-associated protein 31 (Bap31) is an integral membrane protein located in the endoplasmic reticulum (ER) that participates in the transport and quality control of membrane proteins and plays a role in determining cell sensitivity to ER stress and apoptosis. Its cytoplasmic region contains two target sites for caspase cleavage in certain apoptotic pathways. Here, the subcloning, expression, purification and crystallization of the Homo sapiens Bap31 leucine-zipper C-terminal fragment, which spans residues Gly160-Glu246, are reported. An N-terminally His-tagged protein was overexpressed in Escherichia coli and purified by chromatographic methods. X-ray diffraction data were collected in-house to 2.5 A resolution. Crystals belong to space group P6(1)22/P6(5)22, with unit-cell parameters a = b = 70.7, c = 80.6 A. Data analysis indicates the presence of one molecule per asymmetric unit. PMID- 17401200 TI - Overexpression, purification, crystallization and preliminary structural studies of p-coumaric acid decarboxylase from Lactobacillus plantarum. AB - The substrate-inducible p-coumaric acid decarboxylase (PDC) from Lactobacillus plantarum has been overexpressed in Escherichia coli, purified and confirmed to possess decarboxylase activity. The recombinant His(6)-tagged enzyme was crystallized using the hanging-drop vapour-diffusion method from a solution containing 20%(w/v) PEG 4000, 12%(w/v) 2-propanol, 0.2 M sodium acetate, 0.1 M Tris-HCl pH 8.0 with 0.1 M barium chloride as an additive. Diffraction data were collected in-house to 2.04 A resolution. Crystals belonged to the tetragonal space group P4(3), with unit-cell parameters a = b = 43.15, c = 231.86 A. The estimated Matthews coefficient was 2.36 A(3) Da(-1), corresponding to 48% solvent content, which is consistent with the presence of two protein molecules in the asymmetric unit. The structure of PDC has been determined by the molecular replacement method. Currently, the structure of PDC complexed with substrate analogues is in progress, with the aim of elucidating the structural basis of the catalytic mechanism. PMID- 17401201 TI - Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta). AB - Caffeine is a secondary metabolite produced by a variety of plants including Coffea canephora (robusta) and there is growing evidence that caffeine is part of a chemical defence strategy protecting young leaves and seeds from potential predators. The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-L-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). The crystals are orthorhombic, with space group P2(1)2(1)2(1) for XMT and C222(1) for DXMT. X-ray diffraction to 2.8 A for XMT and to 2.5 A for DXMT have been collected on beamline ID23-1 at the ESRF. PMID- 17401202 TI - Purification, crystallization and preliminary X-ray diffraction study on pyrimidine nucleoside phosphorylase TTHA1771 from Thermus thermophilus HB8. AB - Pyrimidine nucleoside phosphorylase (PYNP) catalyzes the reversible phosphorolysis of pyrimidines in the nucleotide-synthesis salvage pathway. In order to study the structure-thermostability relationship of this enzyme, PYNP from the extreme thermophile Thermus thermophilus HB8 (TTHA1771) has been cloned, overexpressed and purified. The TTHA1771 protein was crystallized at 291 K using the oil-microbatch method with PEG 4000 as a precipitant. A native data set was collected to 1.8 A resolution using synchrotron radiation. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 58.83, b = 76.23, c = 103.86 A, beta = 91.3 degrees. PMID- 17401203 TI - Crystallization and preliminary X-ray analysis of the reduced Rieske-type [2Fe 2S] ferredoxin derived from Pseudomonas sp. strain KKS102. AB - The reduced form of BphA3, a Rieske-type [2Fe-2S] ferredoxin component of the biphenyl dioxygenase BphA from Pseudomonas sp. strain KKS102, was crystallized by the sitting-drop vapour-diffusion method under anaerobic conditions. The crystal belongs to space group P3(1)21, with unit-cell parameters a = b = 49.6, c = 171.9 A, and diffracts to a resolution of 1.95 A. A molecular-replacement calculation using oxidized BphA3 as a search model yielded a satisfactory solution. PMID- 17401204 TI - Crystallization of porcine pancreatic elastase and a preliminary neutron diffraction experiment. AB - Porcine pancreatic elastase (PPE) resembles the attractive drug target leukocyte elastase, which has been implicated in a number of inflammatory disorders. In order to investigate the structural characteristics of a covalent inhibitor bound to PPE, including H atoms and the hydration by water, a single crystal of PPE for neutron diffraction study was grown in D(2)O containing 0.2 M sodium sulfate (pD 5.0) using the sitting-drop vapour-diffusion method. The crystal was grown to a size of 1.6 mm(3) by repeated macroseeding. Neutron diffraction data were collected at room temperature using a BIX-3 diffractometer at the JRR-3 research reactor of the Japan Atomic Energy Agency (JAEA). The data set was integrated and scaled to 2.3 A resolution in space group P2(1)2(1)2(1), with unit-cell parameters a = 51.2, b = 57.8, c = 75.6 A. PMID- 17401205 TI - Crystallization and preliminary X-ray diffraction studies of Murraya koenigii trypsin inhibitor. AB - A Kunitz-type trypsin inhibitor purified from the seeds of Murraya koenigii has been crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as the precipitating agent. The crystals belong to the tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 75.8, c = 150.9 A. The crystals contain two molecules in the asymmetric unit with a V(M) value of 2.5 A(3) Da( 1). Diffraction was observed to 2.65 A resolution and a complete data set was collected to 2.9 A resolution. PMID- 17401206 TI - Expression, purification and preliminary X-ray crystallographic studies of the human immunodeficiency virus 1 subtype C protease. AB - Crystals of the human immunodeficiency virus 1 (HIV-1) subtype C protease (PR) complexed with the clinically used inhibitors indinavir (IDV) and nelfinavir (NFV) have been grown in the monoclinic space group P2(1), with mean unit-cell parameters a = 46.7 (+/-0.1), b = 59.8 (+/-0.3), c = 87.0 (+/-0.4) A, beta = 95.2 (+/-0.5) degrees. The crystals of both complexes have been shown to diffract X rays to 2.3 A resolution. The diffraction data for the subtype C PR complexes with IDV and NFV were subsequently processed and reduced, with overall R(sym) values of 8.4 and 11.4%, respectively. Based on the unit-cell volumes, molecular replacement results and packing considerations, there are two protease homodimers per crystallographic asymmetric unit in each of the complexes. The data were initially phased using a model based on the crystal structure of HIV-1 subtype B PR; the structures have been determined and further refinement and analysis are in progress. These structures and subsequent studies with other inhibitors will greatly aid in correlating the amino-acid variation between the different HIV PRs and understanding their differential sensitivity and resistance to current drug therapy. PMID- 17401207 TI - Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of molybdopterin synthase from Thermus thermophilus HB8. AB - Thermus thermophilus is a Gram-negative aerobic thermophilic eubacterium which can grow at temperatures ranging from 323 to 355 K. In addition to their importance in thermostability or adaptation strategies for survival at high temperatures, the thermostable enzymes in thermophilic organisms contribute to a wide range of biotechnological applications. The molybdenum cofactor in all three kingdoms consists of a tricyclic pyranopterin termed molybdopterin that bears the cis-dithiolene group responsible for molybdenum ligation. The crystals of molybdopterin synthase from T. thermophilus HB8 belong to the primitive monoclinic space group P2(1), with unit-cell parameters a = 33.94, b = 103.32, c = 59.59 A, beta = 101.3 degrees. Preliminary studies and molecular-replacement calculations reveal the presence of three monomers in the asymmetric unit. PMID- 17401208 TI - Cloning, expression, purification, crystallization and preliminary crystallographic analysis of selenomethionine-labelled KaiC-like protein PH0186 from Pyrococcus horikoshii OT3. AB - KaiC is the central protein in the circadian-clock system of cyanobacteria. A selenomethionine-labelled KaiC-homologous protein from Pyrococcus horikoshii OT3 (PH0186; 28 kDa) was crystallized by the sitting-drop vapour-diffusion method using ethanol as a precipitant. The crystals diffracted X-rays to beyond 2.0 A resolution using a synchrotron-radiation source. The space group of the crystals was determined to be C2, with unit-cell parameters a = 173.7, b = 51.8, c = 97.5 A, beta = 122.8 degrees. The crystal contains three molecules in the asymmetric unit (V(M) = 2.2 A(3) Da(-1)) and has a solvent content of 43.5%. Sixfold noncrystallographic symmetry was identified from self-rotation calculations, assuming the presence of a hexamer in the crystal. PMID- 17401209 TI - Expression, purification, crystallization and preliminary crystallographic study of isolated modules of the mouse coactivator-associated arginine methyltransferase 1. AB - Coactivator-associated arginine methyltransferase 1 (CARM1) plays a crucial role in gene expression as a coactivator of several nuclear hormone receptors and also of non-nuclear receptor systems. Its recruitment by the transcriptional machinery induces protein methylation, leading to chromatin remodelling and gene activation. CARM1(28-507) and two structural states of CARM1(140-480) were expressed, purified and crystallized. Crystals of CARM1(28-507) belong to space group P6(2)22, with unit-cell parameters a = b = 136.0, c = 125.3 A; they diffract to beyond 2.5 A resolution using synchrotron radiation and contain one monomer in the asymmetric unit. The structure of CARM1(28-507) was solved by multiple isomorphous replacement and anomalous scattering methods. Crystals of apo CARM1(140-480) belong to space group I222, with unit-cell parameters a = 74.6, b = 99.0, c = 207.4 A; they diffract to beyond 2.7 A resolution and contain two monomers in the asymmetric unit. Crystals of CARM1(140-480) in complex with S adenosyl-L-homocysteine belong to space P2(1)2(1)2, with unit-cell parameters a = 74.6, b = 98.65, c = 206.08 A; they diffract to beyond 2.6 A resolution and contain four monomers in the asymmetric unit. The structures of apo and holo CARM1(140-480) were solved by molecular-replacement techniques from the structure of CARM1(28-507). PMID- 17401210 TI - Crystallization and preliminary X-ray crystallographic studies of the biotin carboxyl carrier protein and biotin protein ligase complex from Pyrococcus horikoshii OT3. AB - Biotin protein ligase (BPL) catalyses the biotinylation of the biotin carboxyl carrier protein (BCCP) subunit of acetyl-CoA carboxylase. To elucidate the exact details of the protein-protein interactions in the biotinylation function, the C terminal half fragment of BCCP (BCCPDeltaN76), the R48A mutant of BPL (BPL*) and the R48A K111A double mutant of BPL (BPL**), all of which are from Pyrococcus horikoshii OT3, have been expressed, purified and successfully cocrystallized. Cocrystals of the BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 complexes as well as crystals of BPL*, BPL** and BCCPDeltaN76 were obtained by the oil-microbatch method using PEG 20 000 as a precipitant at 295 K. Complete X-ray diffraction data sets for BPL*-BCCPDeltaN76 and BPL**-BCCPDeltaN76 crystals were collected at 100 K to 2.7 and 2.0 A resolution, respectively, using synchrotron radiation. They belong to the monoclinic space group P2(1), with similar unit-cell parameters a = 69.85, b = 63.12, c = 75.64 A, beta = 95.9 degrees . Assuming two subunits of the complex per asymmetric unit gives a V(M) value of 2.45 A(3) Da( 1) and a solvent content of 50%. PMID- 17401211 TI - Tyrosyl-tRNA synthetase: the first crystallization of a human mitochondrial aminoacyl-tRNA synthetase. AB - Human mitochondrial tyrosyl-tRNA synthetase and a truncated version with its C terminal S4-like domain deleted were purified and crystallized. Only the truncated version, which is active in tyrosine activation and Escherichia coli tRNA(Tyr) charging, yielded crystals suitable for structure determination. These tetragonal crystals, belonging to space group P4(3)2(1)2, were obtained in the presence of PEG 4000 as a crystallizing agent and diffracted X-rays to 2.7 A resolution. Complete data sets could be collected and led to structure solution by molecular replacement. PMID- 17401212 TI - Purification, crystallization and preliminary crystallographic analysis of cytochrome P450 203A1 from Rhodopseudomonas palustris. AB - Cytochrome P450 enzymes constitute a large family of haemoproteins that catalyze the monooxygenation of a great variety of endogenous and exogenous organic compounds. Cytochrome P450 203A1 (CYP203A1, RPA1009) from the metabolically versatile organism Rhodopseudomonas palustris binds a broad range of substrates, in particular substituted aromatic compounds. Crystals of CYP203A1 suitable for X ray crystallography have been obtained and diffraction data were collected in house to 2.0 A resolution from a single crystal. The crystals belong to space group P222, with unit-cell parameters a = 40.1, b = 95.1, c = 99.0 A, alpha = beta = gamma = 90 degrees. There is one protein molecule per asymmetric unit. PMID- 17401213 TI - Expression and purification of F-plasmid RepE and preliminary X-ray crystallographic study of its complex with operator DNA. AB - The replication initiator factor RepE of the F plasmid in Escherichia coli is an essential protein that stringently regulates the F-plasmid copy number. The RepE protein has a dual function: its monomer functions as a replication initiator, while its dimer acts as a transcriptional repressor of the repE gene. The wild type dimeric RepE protein was expressed as an N-terminal histidine-tagged protein, purified under native conditions with a high salt concentration and crystallized in complex with the repE operator DNA using the sitting-drop vapour diffusion technique. The crystals diffracted to a resolution of 3.14 A after the application of dehydration and crystal annealing and belong to space group P2(1), with unit-cell parameters a = 60.73, b = 99.32, c = 95.00 A, beta = 108.55 degrees. PMID- 17401214 TI - Crystallization, preliminary X-ray diffraction and structure analysis of Thermotoga maritima mannitol dehydrogenase. AB - Diffraction data have been collected from a crystal of Thermotoga maritima mannitol dehydrogenase at the Canadian Light Source. The crystal diffracted to 3.3 A resolution and belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 83.43, b = 120.61, c = 145.76 A. The structure is likely to be solved by molecular replacement. PMID- 17401215 TI - Crystallization and preliminary X-ray characterization of phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv. AB - Phosphoglucose isomerase is a ubiquitous enzyme that catalyzes the isomerization of D-glucopyranose-6-phosphate to D-fructofuranose-6-phosphate. The present investigation reports the expression, purification, crystallization and preliminary crystallographic studies of the phosphoglucose isomerase from Mycobacterium tuberculosis H37Rv, which shares 46% sequence identity with that of its human host. The recombinant protein, which was prepared using an Escherichia coli expression system, was crystallized by the hanging-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.8 A and belonged to the orthorhombic space group I2(1)2(1)2(1), with unit-cell parameters a = 109.0, b = 119.8, c = 138.9 A. PMID- 17401216 TI - Purification and crystallization of Vibrio fischeri CcdB and its complexes with fragments of gyrase and CcdA. AB - The ccd toxin-antitoxin module from the Escherichia coli F plasmid has a homologue on the Vibrio fischeri integron. The homologue of the toxin (CcdB(Vfi)) was crystallized in two different crystal forms. The first form belongs to space group I23 or I2(1)3, with unit-cell parameter a = 84.5 A, and diffracts to 1.5 A resolution. The second crystal form belongs to space group C2, with unit-cell parameters a = 58.5, b = 43.6, c = 37.5 A, beta = 110.0 degrees, and diffracts to 1.7 A resolution. The complex of CcdB(Vfi) with the GyrA14(Vfi) fragment of V. fischeri gyrase crystallizes in space group P2(1)2(1)2(1), with unit-cell parameters a = 53.5, b = 94.6, c = 58.1 A, and diffracts to 2.2 A resolution. The corresponding mixed complex with E. coli GyrA14(Ec) crystallizes in space group C2, with unit-cell parameters a = 130.1, b = 90.8, c = 58.1 A, beta = 102.6 degrees, and diffracts to 1.95 A. Finally, a complex between CcdB(Vfi) and part of the F-plasmid antitoxin CcdA(F) crystallizes in space group P2(1)2(1)2(1), with unit-cell parameters a = 46.9, b = 62.6, c = 82.0 A, and diffracts to 1.9 A resolution. PMID- 17401217 TI - Mucin expression profile in Barrett's, dysplasia, adenocarcinoma sequence in the esophagus. AB - BACKGROUND: The molecular events that accompany the progression to adenocarcinoma (ADC) of the esophagus are poorly understood. Aberrant mucin receptor expression can contribute to increased cell growth and metastatic ability. AIM: The aim of this study was to establish a pattern for mucin (MUC) gene expression in the esophageal mucosa under normal and pathological conditions. SETTING: University Hospital Cancer Center Laboratory. Archived tissue samples studied in a retrospective fashion. MATERIALS AND METHODS: Tissue samples were obtained from the archives of patients with histological evidence of Barrett's esophagus (BE) progressing to ADC. Immunohistochemical analysis was performed using mouse monoclonal antibodies for MUC1, MUC2, MUC5AC, MUC6. Semiquantitative scoring of histological staining was performed using a linear scoring system: 0-staining absent; 1-staining in 0-25%; 2-staining in 25-50%; and 3-staining in 50-75% of the epithelium. The Binomial test was used to explore trends and differences in frequency of mucin expression along the pathological sequence. RESULTS: Only mild superficial staining of MUC1 was seen in normal squamous epithelium. MUC1 and MUC2 were uniformly expressed in all samples (7/7) of BE and dysplasia (P=0.008). MUC1 expression was upregulated (7/7) in progression to adenocarcinoma (P=0.008). The secretory mucins, MUC5AC and MUC6 showed a decrease in expression with progression from BE to dysplasia to ADC (P< 0.05). CONCLUSIONS: Downregulation of MUC5AC and MUC6 decreases mucosal protection against gastric acid. Increasing MUC1 expression is associated with progression from dysplasia to ADC. Upregulation of MUC2 reflects intestinal metaplasia in BE. PMID- 17401218 TI - Outcome of patients with stage II and III nonseminomatous germ cell tumors: results of a single center. AB - BACKGROUND: The prognostic factors in nonseminomatous germ cell tumors have been mainly derived from the analysis of stage I tumors. AIMS: The aim of this study was to evaluate some prognostic factors and the outcome of patients with stage II and III nonseminomatous germ cell tumors according to risk groups treated between 1993 and 2002. SETTINGS AND DESIGN: Patients were retrospectively classified as good, intermediate and poor risk groups according to International Germ Cell Cancer Consensus Group. MATERIALS AND METHODS: Biopsy specimens of 58 patients with stage II and III nonseminomatous germ cell tumors were analyzed by means of tumor histopathology, primary localization site of the tumor, relapse sites, initial serum tumor marker levels, the presence of persistent serum tumor marker elevation and the patients' outcome. STATISTICAL ANALYSIS: Kruskall Wallis test and Mann-Whitney U test were used to determine the differences between the groups. Kaplan-Meier method was used for survival analysis and log rank test was used to compare the survival probabilities of groups. Cox proportional hazard analysis was used to determine the prognostic factors in univariate and multivariate analysis. RESULTS: Five-year overall and disease-free survival rates were calculated as 85% and 75% in stage II; 44% and 29% in stage III cases, respectively. Fifty-seven percent of patients were classified in good risk, 9% in intermediate risk and 27% in poor risk groups. Five-year overall survival rates were 97%, 75% and 7% (P<0.001) and disease-free survival rates were 83%, 34% and 7% (P<0.001) in good, intermediate and poor risk groups, respectively. Analysis of the prognostic factors revealed that the localization site of the primary tumor (P<0.001), the initial beta-HCG level (p:0.0048), the presence of yolk sac and choriocarcinoma components in tumor (p:0.003 and p:0.004), relapse sites of tumor (lung versus other than lung) (p:0.003), persistent elevation of serum tumor markers (P<0.001) were significant prognostic factors in univariate analysis. However, in multivariate analysis, only the localization site of tumor (p:0.049) and the relapse site (p:0.003) were found statistically significant. CONCLUSIONS: This retrospective study revealed that in advanced stage of nonseminomatous germ cell tumors, the outcome is essentially related with the localization site of the tumor and the relapse site. PMID- 17401219 TI - Role of radionuclide scintigraphy in the detection of parathyroid adenoma. AB - BACKGROUND: Preoperative detection of parathyroid adenoma is a diagnostic challenge. The sonography and computerized tomography (CT) scan demonstrate high sensitivity but low specificity. The advent of radionuclide scanning technique has enhanced the specificity in this context. AIM: We undertook a study to assess the role of radionuclide scanning in suspected cases of parathyroid adenomas. MATERIALS AND METHODS: Totally 28 cases were incorporated in the study. The suspicion was raised either due to raised PTH levels or recurrent calcinosis. Most of these patients had estimation of calcium done as a routine or specific investigation. The parathyroid scan was performed using either of the two techniques - Dual isotope subtraction or Sestamibi washout technique. We also used the recent approach of fusion imaging (CT + tomographic nuclear images) in selected cases. RESULTS: There were 16 true positive, 10 true negative, 1 false negative and 1 equivocal scan findings. The findings were compared with sonography, CT Scan and PTH values. The true positive yield in our study was 57%, true negative 35% and the overall sensitivity and specificity was found to be 94% and 100% respectively. CONCLUSION: We conclude that parathyroid scintigraphy is a reliable and sensitive technique in the preoperative detection of parathyroid adenomas and should be the first choice of imaging modality in suspicion of parathyroid adenoma. PMID- 17401220 TI - Initiating tumor banking for translational research: MD Anderson and Liverpool experience. AB - The ultimate progress in the cancer diagnosis and therapy has only been possible with the ongoing translational research that is likely to play a very important role in future as well. Hence the importance of such translation from bedside to bench and vis versa cannot be over-emphasized. Accordingly it has become more important to collect tumor samples along with the clinical information in a systematic manner to perform a good basic science research in future. With a population of over a billion and a heavy burden of cancer, India has the 'biggest' potential to establish the 'largest' tumor bank across the globe. Establishing a tumor bank involves money and manpower that may not be feasible across most of the centers in India. Taking into the considering the model of tumor banking of the two leading institutions of the world (MD Anderson Cancer Center, USA and University Hospital Aintree, Liverpool UK), this article presents the salient tips for a center in India to get started with tumor banking with minimal investment. Furthermore a simplified form of ethical consent is presented for the centers to adapt unanimously. PMID- 17401221 TI - Aprepitant: a substance P antagonist for chemotherapy induced nausea and vomiting. AB - The episodes of nausea and vomiting which follow each cycle of chemotherapy are the most troublesome side effect experienced by cancer patients. Introduction of ondansetron was a definite therapeutic advance in treating chemotherapy induced nausea and vomiting (CINV) with more effectiveness with corticosteroids. However, the protection remained largely limited to acute phase of CINV with little or no effect over delayed phase. Aprepitant, a drug that antagonizes the effect of substance P on neurokinin type 1 receptor showed promising results in controlling both phases of CINV. This drug is well absorbed orally with a t max of about four hours. The addition of aprepitant to ondansetron and dexamethasone was found to be superior to ondansetron and dexamethasone alone in clinical trials with patients taking high and moderate emetogenic chemotherapy. This drug also showed a good safety profile, but its inhibitory effect on CYP3A4 may result in clinically significant drug interactions needing dose modifications of co administered drugs. The National Comprehensive Cancer Network guidelines for CINV recommends the use of aprepitant with high and moderately emetogenic anticancer drugs. Results of ongoing clinical trials with aprepitant and other agents of this new class of antiemetics are awaited and may alleviate the sufferings of cancer patients. PMID- 17401222 TI - Cytotoxic drug: towards safer chemotherapy practices. AB - Health care is nearly 10 years behind other industries in its efforts to reduce the errors. Medication error may be nobody's baby, but when it happens, it could well turn out to be everyone's worry and the reasons given for medication error range from silly to the downright serious. The anticancer drugs are known to be mutagenic, teratogenic and carcinogenic, so extra precaution should be taken while storing, diluting, administering the drugs and disposing the waste. The objectives of this article are to define the standards for using cancer chemotherapy in hospitals; to tackle any spillage of drug and how to dispose of the waste of anticancer drugs. This could be beneficial to any hospital where chemotherapy is given without any defined standard operating procedure. The information furnished in this article is collected from the mentioned references and also from websites- The American Cancer Society: Cancer Facts and Figures 2002, www.cancer.org and www.cancersourceRN.com. PMID- 17401223 TI - Frontal convexity primary lymphoma masquerading meningioma: a case report and review of literature. AB - Primary non-Hodgkin's lymphoma of the skull with extra- and intracranial extension without systemic or skeletal manifestation in a nonimmunocompromised patient is extremely rare. These lesions often cause difficulty in diagnosis because they mimic other conditions. We report a case of primary lymphoma involving scalp, skull vault, meninges and invading the brain parenchyma and masquerading clinically and radiologically as a meningioma. PMID- 17401225 TI - Hepatitis E and pregnancy. PMID- 17401226 TI - Outcome of acute liver failure due to acute hepatitis E in pregnant women. AB - INTRODUCTION: Acute liver failure due to acute hepatitis E carries a high mortality. METHODS: Clinical and laboratory parameters of 42 pregnant women (median age 25.5 years) with acute liver failure due to acute hepatitis E were retrospectively analyzed. RESULTS: 22 women delivered, whereas pregnancy continued in 20 women. The maternal mortality in these two groups was similar (9/22 [41%] versus 14/20 [70%], p=0.056). However, in patients with grade I, II or III hepatic encephalopathy, delivery of fetus was associated with reduced mortality in those who delivered as against those who continued pregnancy (5/16 (31%) vs. 13/20 (65%), p=0.046). On multivariate analysis, higher grade of encephalopathy at admission was associated with risk of death (p=0.005). CONCLUSION: Mortality in pregnant women with acute liver failure with acute hepatitis E is high, especially in patients who present with higher grades of encephalopathy. PMID- 17401227 TI - Autoimmune hepatitis in Iranian children. AB - OBJECTIVE: To evaluate the clinical and para-clinical presentation, including autoantibody pattern and response to treatment in Iranian children with autoimmune hepatitis (AIH). METHODS: Data of 60 children presenting with AIH (56 girls) were analyzed retrospectively for clinical, serological and histological profile. RESULTS: Clinical findings included jaundice (67%),hepatomegaly (50%) and ascites (30%). Forty children (38 girls) had type I AIH and 14 (12 girls) had type II AIH. Liver histology was comparable in the two groups and cirrhosis on initial biopsy was equally frequent in Types I and II AIH (63% vs 34%; p=ns). CONCLUSION: Our study describes the presentation and clinical course of Iranian children with AIH. Treatment with corticosteroids and/or azathioprine induces remission of autoimmune hepatitis clinically, biochemically as well as histologically. PMID- 17401228 TI - Salvage surgery in variceal bleeding due to portal hypertension. AB - BACKGROUND: A proportion (10%-15%) of patients with variceal bleeding do not respond to medical management and require surgical intervention. METHODS: Retrospective analysis of 82 consecutive patients (median age 31 years, range 3 71; 60 male) who underwent salvage surgery for variceal bleeding between 1989 and 2005. RESULTS: Immediate control of variceal bleeding was achieved in 78 (95%) patients. Four patients (cirrhosis 3, portal vein block 1) continued to bleed in the postoperative period following gastro-esophageal devascularization (3) or portacaval shunt (1). Twelve (15%) patients died in hospital; the commonest cause of death (n=7) was liver failure and sepsis leading to multi-system organ failure. The mortality rate was higher among patients who had undergone emergency surgery for active bleeding than among those who had been adequately resuscitated and evaluated prior to surgery (12/45 vs. 0/37; p< 0.001). Mortality rate tended to be higher in patients with cirrhosis (overall 10/45 [22%]; Child A 1/9 [11%], Child B 4/20 [20%], Child C 5/16 [31%]) than in those with non-cirrhotic portal hypertension (3/37 [8%]; p=ns). CONCLUSIONS: Our data suggest that salvage surgery is justified in patients with variceal bleed in whom non-surgical measures fail. PMID- 17401229 TI - Effect of cimetidine on hepatotoxicity induced by isoniazid-rifampicin combination in rabbits. AB - OBJECTIVE: To evaluate the hepatoprotective potential of cimetidine in hepatotoxicity induced by isoniazid-rifampicin combination in albino rabbits. METHODS: Six groups of six rabbits each were studied. Three groups received saline (control), isoniazid (50 mg/Kg/d) alone or isoniazid with rifampicin (100 mg/Kg/d) daily orally for 7 days. Other groups received intraperitoneal cimetidine (50 mg/Kg/d) alone or cimetidine (50 or 120 mg/Kg/d) along with isoniazid-rifampicin combination. Serum levels of liver enzymes were measured at baseline and on day 8 and liver histology was studied on day 8. RESULTS: Rabbits receiving isoniazid alone for 7 days showed no increase in serum ALT and AST levels, whereas those receiving isoniazid-rifampicin combination had a 3-4-fold increase in these levels (p=0.02). Animals receiving cimetidine pre-treatment did not show a significant increase in ALT and AST levels. Histological changes in the liver were more common with isoniazid-rifampicin combination than with isoniazid only. These changes were reduced in animals receiving low-dose cimetidine and prevented in those receiving high-dose cimetidine. CONCLUSION: Cimetidine in high dose can prevent hepatotoxicity induced by isoniazid rifampicin combination. PMID- 17401230 TI - Jejunal duplication cyst: diagnosis aided by scintigraphic detection of ectopic gastric mucosa. PMID- 17401231 TI - Determining standard liver volume: assessment of existing formulae in Indian population. AB - BACKGROUND: With the increasing numbers of living-related donor liver transplantation, accurate means of calculating standard liver volume (SLV) based on patient body indices becomes important. Three formulae reported in literature for this purpose have been derived from studies on Western and Japanese populations. AIM: To assess the existing formulae for calculation of SLV in Indian population. METHOD: Total liver volume (TLV) of 238 patients was measured using axial helical CT images obtained for conditions unrelated to the hepatobiliary system. Body surface area (BSA) was calculated from height and weight. Measurements obtained using CT were compared with the SLV calculated based on the previously reported formulae. RESULTS: Though there was significant difference (p< 0.001) between the TLV obtained by CT and the SLV calculated using the three formulae, they also showed good agreement. On an average the formula derived from the Japanese population underestimated the SLV by 63 (202) cc (p< 0.001). Regression models for SLV (SLV = 243 + [186 x BSA] + [11.4 x Weight], SLV = 375.23 + [14.24 x body weight], SLV = -204.092 + [874.461 x BSA]) were derived from the data obtained from our population. Age and gender had no effect on the SLV. CONCLUSIONS: Formulae derived from Japanese population for calculation of SLV is not suitable for the Indian population. The newly described formulae may prove useful in the Indian population. PMID- 17401232 TI - Current therapeutic options for colorectal liver metastases. PMID- 17401233 TI - Jaundice and firm hepatomegaly in an irritable male child. PMID- 17401234 TI - Outcome of radical excision of anorectal gastrointestinal stromal tumors in Hong Kong Chinese patients. AB - Anorectal gastrointestinal stromal tumor (GIST) is a rare disease entity with malignant potential. Medical records of six patients (median age 68 years) with anorectal GIST who underwent surgical treatment at our institution between 1992 and 1999 were retrospectively reviewed. Four patients presented with rectal bleeding. The tumors were located in the mid and lower rectum in 4 patients and in the anal canal in 2 patients. The median tumor diameter was 4.5 cm. One patient who had undergone local excisions in another hospital presented with recurrent GIST. He refused radical excision and underwent wide local excision again. He developed recurrence 2 years later and underwent salvage pelvic exenteration, but finally died of disseminated disease. Five patients underwent initial radical excision. Among them, 3 developed recurrences (one each local, distant and both) at a median duration of 50.3 months. Two patients died of the disease, while one patient who had both local and distant recurrences resected remained alive till the end of the study period (median duration of follow-up of the 5 patients was 84.6 months). At 5 years, of 5 patients who underwent initial radical excision, 3 and 4 patients, respectively, had disease-free and overall survival. Recurrence of anorectal GIST is common despite radical excision. Nevertheless, a reasonable survival rate can be achieved. PMID- 17401235 TI - Recurrent limb weakness as initial presentation of Wilson's disease. AB - A 28-year-old normotensive euthyroid man presented with recurrent lower motor neuron type of weakness without sensory or autonomic involvement, with preserved reflexes. Systemic examination was significant for mild hepatosplenomegaly. Investigations revealed persistent hypokalemic, hyperchloremic, normal-anion-gap metabolic acidosis with deranged liver functions. Urine pH was 6.0 even after oral ammonium-chloride loading test. Type I renal tubular acidosis was diagnosed. A search for the etiology revealed bilateral Kayser-Fleischer ring, with low serum ceruloplasmin levels and high urinary copper, confirming it to be Wilson's disease. PMID- 17401236 TI - Sebaceous glands in esophagus in gastroesophageal reflux disease. AB - Sebaceous glands in the esophagus are very rare and have been reported mostly in autopsy studies. They have been considered to be of no clinical significance. We report a 50-year-old man with gastroesophageal reflux disease who had sebaceous glands in the esophagus on endoscopic biopsy. PMID- 17401237 TI - Double-barrel common bile duct: a rare cause of extrahepatic biliary obstruction. AB - Double-barrel common bile duct is rare. We report a 50-year-old woman with defective canalization of the common bile duct, presenting with extrahepatic biliary obstruction due to stones in one compartment. CT scan highlighted this anomaly. After failed attempts at stone extraction at ERCP, she was successfully operated on. PMID- 17401238 TI - Adult idiopathic ileosigmoid intussusception prolapsing per rectum. AB - Ileosigmoid intussusception can lead to ischemia and necrosis of either the ileum or sigmoid colon. Ileosigmoid intussusception as a mass prolapsing per rectum in an adult has not been previously reported. We report a 50-year-old man with such a presentation. He recovered uneventfully after subtotal colectomy. PMID- 17401239 TI - Esophageal stricture following electrical injury. AB - Esophageal injury resulting from electrical shock is rare. Stricture of esophagus following external electrical injury has not been reported yet. We report a 24 year-old electric lineman who developed esophageal stricture following external electrical shock. He responded to dilatation with Savary-Gilliard dilators. PMID- 17401240 TI - Coexistence of chronic calcific pancreatitis and celiac disease. AB - Coexistence of celiac disease with chronic calcific pancreatitis is rare. We describe a 26-year-old woman with chronic calcific pancreatitis in whom non response to treatment was due to celiac disease. PMID- 17401241 TI - Landry-Guillaine-Barre syndrome as presentation of celiac disease. AB - Celiac disease has been associated with a variety of neurological illnesses, most frequently cerebellar ataxia and peripheral neuropathy. We report presentation as Landry-Guillaine-Barre syndrome in a 28-year-old woman with previously unsuspected celiac disease. PMID- 17401242 TI - Massive recurrent hemorrhage due to duodenal diverticulum--role of tattooing and suture plication. AB - We report a 79-year-old lady with massive recurrent GI bleeding from a duodenal diverticulum arising at the mesenteric border. We used tattooing during endoscopy as a method for identifying the bleeding source at surgery. The diverticulum was treated with suture plication. PMID- 17401243 TI - Serum and ascitic fluid CA-125 level in patients with ascites. PMID- 17401244 TI - Concordance between endoscopic and histological gastroesophageal reflux disease. PMID- 17401245 TI - Prevalence of celiac disease in first-degree siblings of celiac disease patients. PMID- 17401246 TI - Randomized, comparative study of cefotaxime 2 versus 4 grams in spontaneous bacterial peritonitis. PMID- 17401247 TI - Persistence of anti-HBs titers after two different doses of Genevac B, a recombinant hepatitis B vaccine, in healthy adolescents. PMID- 17401248 TI - Is laparoscopic appendectomy contraindicated in mucocele of appendix? PMID- 17401249 TI - Pitfalls in the diagnosis of insulinoma. PMID- 17401250 TI - CT colonography versus conventional colonoscopy. PMID- 17401251 TI - Ciprofloxacin-induced cholestatic jaundice. PMID- 17401252 TI - Predicting mortality in critically ill obstetric patients requiring intensive care unit admission in India. PMID- 17401253 TI - Closed-system suctioning: why is the debate still open? PMID- 17401254 TI - Prediction of mortality and morbidity by simplified acute physiology score II in obstetric intensive care unit admissions. AB - BACKGROUND: Critical care in obstetrics has received much attention in recent times. Despite progress in medical field and improvement in health facilities provided, maternal mortality is still very high in most of the developing countries. AIM: To study and analyze records of patients requiring intensive care in obstetrics and to assess utility of simplified acute physiology score (SAPS II) for predicting maternal mortality. SETTING: A multidisciplinary intensive care unit (ICU) at a tertiary care center. DESIGN: Retrospective review. MATERIALS AND METHODS: Fifty-seven consecutive obstetric patients' records requiring ICU admissions were studied for clinical picture, diagnosis, complications, morbidity and mortality over a period of 21/2 years - from 1st May 2002 to 31st Oct. 2004. SAPS II score was calculated according to the different variables for predicting mortality. STATISTICAL ANALYSIS: SAPS II scores were regressed on mortality status using logistic regression analysis. The predictability was assessed by goodness-of-fit test and receiver operated characteristic curve. RESULTS: Maternal mortality in obstetric ICU admissions was 1.15/1,000 deliveries, amounting to 40.35% of obstetric ICU admissions. The mean SAP II score was significantly higher (40.04 +/- 12.97 vs. 22.6 +/- 7.31) in those patients who died compared to survivors (P < 0.001%). CONCLUSIONS: The SAPS II accurately predicted mortality in obstetric patients admitted to ICU. Computation of the score as a routine in ICU may help in identifying those at high risk of mortality and then to reduce this risk. PMID- 17401255 TI - Post-sterilization regrets in Indian women. AB - BACKGROUND: Tubal ligation is the most common form of contraception in India. We conducted this study to assess the factors associated with post-sterilization regrets. AIMS: This study was designed to assess risk factors that are likely to cause regret following female sterilization in Indian women. SETTINGS AND DESIGN: Questionnaire-based study. MATERIALS AND METHODS: This was a questionnaire-based study to assess the level of satisfaction after tubal ligation in women attending family planning clinic at the hospital over a period of 1 year. Evaluation of data obtained through questionnaire from 236 women who had undergone tubal ligation was done. STATISTICAL ANALYSIS: Univariate analysis to determine crude odds ratio was carried out. Subsequently, multiple regression analysis was used to find the adjusted odds ratio (and 95% confidence intervals) for each variable. RESULTS: We found a strong co-relation between regrets and young age (less than 30 years), fewer number of children, few or no male children and lack of partner motivation prior to sterilization. Menstrual irregularities and dysmenorrhoea did not influence regret to a large extent. CONCLUSION: Fertility-related factors, namely, age at sterilization, family size, number of male offspring, timing of sterilization and non-involvement of partner in decision making played a greater role in post-sterilization regrets than menstrual factors (menstrual irregularities and dysmenorrhoea). PMID- 17401256 TI - Private sector contribution to childhood immunization: Sri Lankan experience. AB - BACKGROUND: The main service provider for childhood immunization in Sri Lanka is the government sector. However, utilization of private sector for childhood immunization is increasing rapidly. Existing national immunization data does not routinely include statistics on private sector immunization delivery adequately. OBJECTIVE: To estimate the proportion of children immunized in the private sector; describe socio-demographic characteristics of private sector users and compare these with government sector users. MATERIALS AND METHODS: A community based crosssectional descriptive study was conducted using a pre-tested interviewer-administered structured questionnaire. This was done in the Colombo municipal council area using the WHO 30 cluster methodology. The total number of households in the sample was 553. RESULTS: Out of the 5,028 total immunizations reported in the present study, around one-third (2,544) was obtained through the private sector. Nineteen percent (104) of children were exclusively immunized from the private sector. The distribution of usual immunization provider was - government sector 72.3% (400) and private sector 27.7% (153). Significant differences were observed (P < 0.001) between private and government sector users with regard to family income, social class, ethnicity, religion and educational level of the mother. The age-appropriate immunization among the 12- to 23-month age group was 92.3% (144) in the government sector, whereas it was 95% (38) in the private sector. Among the 24- to 35-month age group, it was 91.7% (121) and 92.7% (76) respectively. The age-adjusted immunization coverage rates were almost same among the government and private sector users except for the measles vaccine, where the private sector users had significantly (P = 0.016) higher coverage. CONCLUSIONS: Utilization of private sector immunization services is high in the Colombo municipal council area. PMID- 17401257 TI - Comparison of closed endotracheal suction versus open endotracheal suction in the development of ventilator-associated pneumonia in intensive care patients: an evaluation using meta-analytic techniques. AB - BACKGROUND: Ventilator-associated pneumonia (VAP), a frequent nosocomial infection in the intensive care, is associated with considerable morbidity. Endotracheal suctioning is routinely performed in mechanically ventilated patients to clear secretions. This study assessed if there were advantages of closed endotracheal suctioning (CES) over open endotracheal suctioning (OES) with respect to clinical outcomes. MATERIALS AND METHODS: Trials comparing CES with OES were identified by search of MEDLINE (1966-July 2006) and bibliographies of relevant articles. Only trials reporting VAP and/or mortality were considered. Studies reporting only physiological outcomes were excluded. STATISTICAL ANALYSIS USED: A meta-analysis of randomized controlled trials (RCTs) was performed using the random-effects estimator. The effect of suctioning type on VAP and mortality was reported as risk difference (RD) and duration of mechanical ventilation (MV) as mean weighted difference (MWD). RESULTS: Nine RCTs fulfilled criteria for inclusion. There was no differential treatment effect of suctioning type (closed versus open, n = 9 studies) on VAP (RD - 0.01; 95% CI - 0.05, 0.03; P = 0.63) or on mortality (n = 5; RD 0.01; 95% CI - 0.04, 0.05; P = 0.8). Although OES was associated with a shorter duration of MV (n = 4; MWD -0.64; 95% CI 0.21, 1.06; P = 0.004), one study contributed significantly to the estimates. Heterogeneity of treatment effects was not observed. CONCLUSIONS: This meta-analysis has not demonstrated a superiority of CES over OES with respect to VAP or mortality. Thus the decision for the use of CES may be based on possible benefits in patients requiring high respiratory supports, reduced costs in those needing prolonged MV or occupational health and safety concerns with OES. PMID- 17401258 TI - Acro-osteolysis and mononeuritis multiplex as a presenting symptom of systemic angiitis of Wegener's type. AB - Wegener's granulomatosis is a multisystem disorder involving small- and medium sized vessels, leading to granuloma formation and involvement of upper and lower respiratory tract with or without glomerulonephritis. However, limited forms of angiitis and granulomatosis of the Wegener's type with oligosymptomatic and atypical site involvement are known to occur. We present here a rare case of limited form of angiitis and granulomatosis of Wegener's type who presented sequentially with spontaneous resorption of digits with acro-osteolysis and mononeuritis multiplex over a period of 10 months. His vasculitic workup revealed high proteinase 3 antibodies (c-ANCA) titers and an almost asymptomatic lung involvement, detected on high-resolution computed tomography of chest. The patient was aggressively treated with immunosuppressive therapy, following which he showed good improvement. PMID- 17401259 TI - Personality characteristics and gastroesophageal reflux disease in southern Iran. PMID- 17401260 TI - Toxoplasma myocarditis presenting as myocardial infarction. PMID- 17401261 TI - Information and communication technology developments in asthma management: a systematic review. AB - This review aims to explain the progress of information and communication technology (ICT) applications in asthma management. Appropriate literature was printed out from the bibliographic databases and library source using relevant key phrases of ICT and asthma. The ICT developments from simple to complex modules to augment the conventional methods of asthma care with a caution of excessive reliance upon technology were discussed. However, it should be noted ICTs are for maximizing the human clinician's own ability to receive and process information as well as providing unique opportunities for patients, physicians, pharmacists and researchers. PMID- 17401262 TI - Newborn care in rural Uttar Pradesh. AB - OBJECTIVES: To describe selected newborn care practices related to cord care, thermal care and breastfeeding in rural Uttar Pradesh and to identify socio demographic, antenatal and delivery care factors that are associated with these practices. METHODS: A cross-sectional survey in rural Uttar Pradesh included 13,167 women who had a livebirth at home during the two years preceding data collection. Logistic regression was used to identify socio-demographic, antenatal and delivery care factors that were associated with the three care practices. RESULTS: Use of antenatal care and skilled attendance at delivery were significantly associated with clean cord care and early breastfeeding, but not with thermal care. Antenatal home visits by a community-based worker were associated only with clean cord care. Women who received counseling from health workers or other sources on each of the newborn care practices during pregnancy were more likely to report the respective care practices, although levels of counseling were low. CONCLUSION: The association between newborn care practices and antenatal care, counseling and skilled delivery attendance suggest that evidence-based newborn care practices can be promoted through improved coverage with existing health services. PMID- 17401263 TI - Vinyl bags prevent hypothermia at birth in preterm infants. AB - OBJECTIVE: To compare the effect of standard care vs. the use of vinyl bags (Vi Drape (R) isolation bag) on admission temperature in extremely premature infants < or = 28 weeks gestational age at birth. METHODS: Twenty seven premature infants with similar baseline characteristics were assigned and placed in vinyl bags (n=14) immediately following delivery without drying or received standard care (controls, n=13), including drying and placement under a radiant warmer. Axillary temperature was recorded on admission to the neonatal unit. RESULTS: The average temperature in the vinyl bag group was significantly higher (35.9 +/- 0.13 vs 34.9 +/- 0.24 degrees C, p=0.002). Although the cord blood pH was similar between the two groups (7.33+0.02 in the vinyl bag group and 7.33 +/- 0.01 in the control group), the worst pH in the first 6 hours of life was significantly lower in the control group (7.32 +/- 0.02 vs 7.22 +/- 0.04, p=0.03). There was a significant increase in maximal oxygen requirement during the first 24 h in the control group (82.9 vs 43.3% in the vinyl bag group, p=0.0004). CONCLUSION: Vinyl bags prevent heat loss and are a simple and effective intervention in preventing hypothermia in the delivery room and early acidosis in premature infants. PMID- 17401264 TI - Prognostic cytogenetic markers in childhood acute lymphoblastic leukemia. AB - OBJECTIVE: To evaluate children with acute lymphoblastic leukemia (ALL) showing resistance to immediate induction chemotherapy in relation to conventional and advanced cytogenetic analysis. METHODS: This work was conducted on 63 ALL children (40 males and 23 females) with age range 4.5 months-16 years (mean = 7.76 years). They included 37 cases attained true remission and 26 complicated by failure of remission, early relapse or death. They were subjected to history, clinical examination and investigations including CBC, BM examination, karyotyping, FISH for translocations and flowcytometry for immunophenotyping and minimal residual disease diagnosis. RESULTS: Cases aged 50.000/mm3 also showed better but non-significant remission rates. Most of the present cases were L2 with better remission compared to other immunophenotypes. Forty informative karyotypes were subdivided into 15 hypodiploid, 10 pseudodiploid, 8 normal diploid and 7 hyperdiploid cases; the best remission rates were noticed among the most frequent ploidy patterns. Chromosomes 9, 11 and 22 were the most frequently involved by structural aberrations followed by chromosomes 5, 12 and 17. Resistance was noted with aberrations not encountered among remission group; deletions involving chromosomes 2p, 3q, 10p and 12q; translocations involving chromosome 5; trisomies of chromosomes 16 and 21; monosomies of 5 and X and inversions of 5 and 11. CONCLUSION: Some cytogenetic and molecular characterizations of childhood ALL could add prognostic criteria for proper therapy allocation. PMID- 17401265 TI - Obesity and related risk factors. AB - OBJECTIVE: To study the prevalence of overweight and obesity among Iranian schoolgirls and to identify risk factors which lead to obesity. METHOD: This cross-sectional study was conducted in 2002 and a sample of 1800 female students between 7-12 years old was obtained using a multistage cluster sampling method from Tehran. Height and weight were measured and related socio-economic information was collected. RESULTS: The overall percent of overweight and obesity was 13.3% and 7.7% respectively. BMI (Body Mass Index) was directly and significantly(r=+0.28, P< 0.001) correlated with increasing age. Physical activity was significantly different between obese and non-obese children. (P=0.03) Also, economical factors such as the type of school (private&public) were different in these children. (P=0.03) The statistical analysis of the data revealed a significant and inverse correlation(r=-0.03, P=0.04) between maternal education and occurrence of overweight and obesity in children. CONCLUSION: The prevalence of overweight and obesity in young Iranian girls was high. Advanced age, lack of physical inactivity, low economical factors and maternal educational status could be risk factors for obesity in children. PMID- 17401266 TI - Thrombocytopenia related neonatal outcome in preterms. AB - OBJECTIVE: To investigate the thrombocytopenia and platelet transfusion related outcome in very preterm infants. METHODS: Cases (n=94) with at least one episode of thrombocytopenia (platelet counts < 150 x 10(9)/L) and controls (n=70) were identified from a database of 1054 neonates with gestational age < or = 32 weeks admitted to a level III NICU. Thrombocytopenia and platelet transfusion related morbidity (IVH, sepsis, NEC, and bleeding) and mortality were analyzed with respect to gestational age (< 28 weeks and 28-32 weeks), severity of thrombocytopenia (mild if platelet count > or = 100 and < 150 x 10(9)/L, moderate if count > or = 50 and < 100 x 10(9)/L, and severe if platelets < 50 x 10(9)/L), age of thrombocytopenia onset (early < 72 hours and late > or =72 hours). RESULTS: The majority of thrombocytopenia (67.0%) was diagnosed after 72 hours of age, and was mild in 12.8%, moderate in 36.2% and severe in 51.0% of the cases. Neonates with severe and moderate thrombocytopenia were more frequently born at lower gestational age and birth weight. NEC and sepsis especially that caused by Candida infection, were associated with severe thrombocytopenic events. The development of IVH was strongly associated with lower gestational age but not the severity and age of thrombocytopenia onset. Mucocutaneous bleeding complicated 18.4% of cases with severe and late-onset thrombocytopenia (7/38). Platelets were transfused to 85.4% of infants with severe and 64.7% of infants with moderate thrombocytopenia (P< 0.02). The gestational age of the majority of the platelet transfused neonates (49/60, 81.7%) was < 28 weeks. Mean gestational age and birth weight, and rates of severe thrombocytopenia, IVH, sepsis and mortality were comparable in transfused vs not-transfused infants with gestational age 28-32 weeks. Platelet transfused neonates with gestational age < 28 weeks had lower birth weights, were more often severely thrombocytopenic, and died more frequently than infants of a similar gestational age who were not transfused. CONCLUSION: Platelet transfusions did not lower mortality in very premature born infants with moderate and severe thrombocytopenia during the NICU admission. PMID- 17401267 TI - Quality of life issues in caregivers of youngsters with thalassemia. AB - OBJECTIVE: To assess the contribution of psychiatric problems, concerns regarding the illness to Quality of Life (QOL) in caregivers. METHODS: Forty four caregivers were screened for psychiatric problems using the General Health Questionnaire 12 item version (GHQ-12) and their concerns and QOL were assessed using the Concerns Checklist and the EQ5D respectively. Clinical psychiatric interview was conducted for those qualifying for GHQ caseness and a psychiatric diagnosis was ascribed. These were correlated with the Childhood Psychopathology Measurement Schedule/ GHQ- 12 scores (as appropriate) and the EQ5D score of the youngster. RESULTS: Fifty seven percent of the caregivers had psychiatric problems, with depressive disorders in 19/44 (45%) QOL was affected in 50% (Pain/ discomfort (n =19, 45%), Mobility (n = 7, 17%), Depression and Usual activities dimensions (n= 6, 14% each). The greatest concerns were regarding the future (91%), illness (80%) and finances (73%). CONCLUSION: Caregiver support through psychological interventions or medication would alleviate psychiatric problems and concerns and contribute towards a better QOL. PMID- 17401268 TI - Safety and efficacy of rifampicin in children with cholestatic pruritus. AB - OBJECTIVE: The present study aimed at verifying the safety and efficacy of rifampicin in ameliorating pruritus in cholestatic children. METHODS: Twenty three Egyptian children (14 boys and 9 girls), suffering from intractable pruritus of cholestasis, were included. Rifampicin was started at a dose of 10 mg/Kg/day in two divided doses and increased gradually to a maximum of 20 mg/Kg/day if there was no response. Liver function tests were followed up weekly. RESULTS: Seventeen patients (74%) showed improvement of pruritus with rifampicin. None of the patients showed any deterioration in liver functions. CONCLUSION: Rifampicin in a dose of 10-20 mg/Kg/day is safe and effective in ameliorating uncontrollable pruritus in children with persistent cholestasis. PMID- 17401269 TI - Life on streets. AB - OBJECTIVE: Millions of children leave home in hope of finding a more tolerable life. The study tries to find out the kind of life they lead once out on the streets. METHODS: Cross-sectional study of all street boys admitted to a Child Observation Home in New Delhi during specified 6 months. Questions regarding various aspects of survival on streets were asked. RESULTS: Majority are forced to work in informal sector for survival with earnings well below the minimum wages. To escape from the exploitation and hardships, many begin abusing substances. The street experience leads to hopelessness for the future and suicidal tendencies. CONCLUSION: After circumstances at home force these kids to escape, they are caught up in yet another world of exploitation, hunger, violence and abuse. PMID- 17401270 TI - Epidemiology and pathophysiology of Helicobacter pylori infection in children. AB - Helicobacter pylori is one of the commonest bacterial pathogens in human. The organism is associated with development of peptic ulcer diseases, lymphoproliferative disorders and gastric cancer. Residence in a developing country, poor socio-economic conditions and genetic predisposition are regarded as risk factors. Prevalence of infection is higher in developing countries and re infection is higher among under five children. It is transmitted mainly through feco-oral route in developing countries and gastro-oral route in developed nations. Transmission of 'close-contact infection' depends on the degree of mixing and age-distribution between susceptible and infected individuals. Host and bacterial factors with interaction of environment contribute pathogenicity. H. pylori cytotoxin-associated geneA (cagA), vacuolating toxinA (vacA) and adherence factors to gastric epithelium have been linked to enhanced pathogenicity of the bacterium. Host genetic polymorphism of cytokines, related legends, receptors and enzymes influence H. pylori infection. PMID- 17401271 TI - Ghosal type hemato-diaphyseal dysplasia: a rare variety of Engelmann's disease. AB - Ghosal type hemato-diaphyseal dysplasia is a recently described clinical entity. The authors describe such a case with severe anemia requiring transfusions and with clinical and radiological evidence of diaphyseal dysplasia. Very few such cases are reported in world literature. PMID- 17401272 TI - Arachnoid cyst with growth hormone deficiency. AB - On clinical grounds, arachnoid cysts are usually associated with neurological dysfunction. Little is known concerning their involvement in endocrine disorders. A seven-year-old boy was admitted to the hospital for evaluation of an unprovoked afebrile seizure. His neurological examination was normal, however, he had growth retardation. Insulin tolerance and L-dopa growth hormone stimulation tests revealed an inefficient growth hormone response. An MRI of hypophysis and cranium yielded a shift of hypophysis and a large arachnoid cyst. PMID- 17401273 TI - Spinal tuberculosis mimicking malignancy: atypical imaging features. AB - Here is reported a 14-year-old boy with spinal tuberculosis. The imaging features were suggestive of non hodgkin's lymphoma with sparing of intervertebral disc. The atypical imaging features of spinal tuberculosis is discussed. PMID- 17401274 TI - Spontaneous infantile choledochal cyst perforation. AB - Spontaneous perforation is a rare complication of infantile choledochal cysts. A 23-weeks female with acute biliary peritonitis is described and differentiation from a spontaneous perforation of the extrahepatic bile duct highlighted. PMID- 17401275 TI - Gastropleural fistula as a complication of empyema thoracis. AB - Gastropleural fistula is an uncommon entity, especially in children. Here we report a 7-year-old child who developed gastropleural fistula as a complication of empyema thoracis. The child was also diagnosed to have chronic granulomatous disease. PMID- 17401276 TI - Lung hypoplasia with dilated cardiomyopathy: coincidence or association? AB - Pulmonary hypoplasia or aplasia is part of the spectrum of malformations characterized by incomplete development of lung tissue. In more than 50% of these cases, coexisting cardiac, gastrointestinal, genitourinary, and skeletal malformations are present, as well as the variations in bronchopulmonary vasculature. In literature there is no reported case describing lung hypoplasia with dilated cardiomyopathy without structural heart disease. Here, it is presented a 6-month girl, referred to us for persistent homogenous opacity in left hemithorax detected on chest X-ray. Subsequently, she was found to have dilated cardiomyopathy without structural congenital heart disease and congenital hypoplasia of left lung. PMID- 17401277 TI - Pseudoaneurysm of internal carotid artery. AB - Pseudoaneurysms of the extracranial Internal Carotid Artery (ICA) are rare. Here it is reported a case of posttraumatic extracranial ICA pseudoaneurysm in a three year-old boy. The pseudoaneurysm arising from the extracranial ICA was initially diagnosed by DSA. Later on confirmed by Doppler and MRA. The imaging features are described with a brief review of literature. PMID- 17401278 TI - Congenital paraesophageal hernia presenting with severe gastroesophageal reflux. AB - Paraesophageal hernia in children is relatively rare entity. These children usually present with recurrent respiratory tract infection or vague GI symptoms. An 11 year female presented with episodic vomiting, heartburn and features of hypocalcemic tetany. Investigations revealed right paraesophageal hernia. Reduction of the contents and repair of the defect with fundoplication was done. Patient is asymptomatic on follow-up. PMID- 17401279 TI - Unilateral pseudohypacusis in children. AB - Pseudohypacusis is a condition where there is hearing loss without any organic etiology. Though this condition has been reported long before, the literature exists mainly in the form of case reports. An interesting aspect of pseudohypacusis is of patients presenting with unilateral pseudohypacusis. In children, there often exist underlying psychological factors related to the occurrence of this condition. This condition is usually suspected when there is discrepancy in audiometric testing. The authors hereby report a child of unilateral pseudohypacusis presenting with psychological stressors, along with an overview of the management of such conditions. PMID- 17401280 TI - Vitreous hemorrhage secondary to vitamin K deficiency bleeding. PMID- 17401281 TI - [Evaluation of psychotherapy treatments]. PMID- 17401282 TI - [Evaluation of psychotherapy treatments]. PMID- 17401283 TI - [Walking activity in prosthesis-bearing lower-limb amputees]. AB - PURPOSE OF THE STUDY: A large body of literature has been devoted to gait analysis in amputees. Most studies have been conducted in the laboratory setting where numerous variables are analyzed: gait efficiency, energy cost, walking velocity. At the present time however, data are lacking on the real-life use of walking prostheses. Little is known about how long patients wear their prosthesis or how difficult it is for them to walk during different periods of the day. Currently, such information depends on the quality of the interview and the patient-physician relationship. A precise assessment of walking activity in amputees and use of prostheses would be a valuable source of information for therapists and would provide complimentary information to that collected from laboratory gait analysis. The purpose of this study was to study walking activity in home-dwelling prosthesis-bearing lower-limb amputees and to determine variables affecting walking performance. To our knowledge, this is the first published study on this topic. MATERIAL AND METHODS: The series included home dwelling prosthesis-wearing lower-limb amputees (Syme amputation or more proximal) who were able to get up and go without assistance. A StepWatch3 recorder was implanted on the prosthesis for 15 consecutive days. Variables recorded were number of steps, total walking time, and walking velocity. Variables which might affect walking in prosthesis-wearing amputees were also recorded: body mass index, use of a walking aid, level and reason for amputation, age at amputation and at recording, time between amputation and recording. RESULTS: From June 2004 to May 2005, 43 patients wore the StepWatch3 for the scheduled 15 days during their daily activities. Mean age at amputation was 42 years (range 13-78 years) and at recording 52 years (range 25-85 years). Considered separately, gait parameters showed that all of the patients wore their prosthesis daily and that the best walking performance was significantly observed among below-knee amputees who did not use a walking aid and who underwent amputation for a non-vascular cause. Multivariate analysis revealed that above knee amputees lost 93 minutes of walking time per day (21% loss), and that amputees who used a walking aid lost 58 minutes per day (13% loss). Daily walking time declined 2.5 minutes per year of age. Body mass index was not correlated with total daily walking time but had a direct significant effect on walking velocity. PMID- 17401284 TI - [Acromio-clavicular dislocations treated by synthetic coraco-clavicular ligamentoplasty]. AB - PURPOSE OF THE STUDY: The aim of this study was to evaluate results of coraco clavicular ligamentoplasty using a synthetic material, dacrylene, for the treatment of recent and neglected acromio-clavicular dislocation. MATERIAL AND METHODS: The series initially included 104 patients who underwent surgery from 1981 to 2001. Among these, 75 were reviewed with a mean follow-up of 85 months (72% review rate). A complete set of clinical (Constant score/100 points) and radiographic (coraco-clavicular distance) data was available for 39 patients. Thirty-six answered a questionnaire with measurement of a partial Constant score (65 points). RESULTS: Seventy-seven percent of patients were satisfied or very satisfied and 98% presented good or very good functional outcome as assessed by the partial Constant score. Reduction of the coraco-clavicular distance was satisfactory for 54% of patients evaluated. For the vast majority, insufficient reduction was reducible and had no impact on the functional outcome. The time to treatment (recent or neglected dislocation) had no effect on patient satisfaction or functional outcome. The coraco-clavicular distance was greater for dislocations operated on late (p<0.05). The grade of the injury, the type of assembly used, and use or not of lateral clavicular resection had no effect on clinical or radiological outcomes. Dacrylene was tolerated poorly by 14 patients (18.7%). The functional outcome was very good for all after removal of the ligament. DISCUSSION: Functional results are good with this technique which enables a well sustained stable reduction of recent and neglected dislocations, irrespective of the grade of the lesion or the type of assembly used. Reducing the mechanical stress on the ligament and prudent rehabilitation exercises should be helpful in preventing intolerance reactions. PMID- 17401285 TI - [Treatment of chronic anterior shoulder instability using a coracoid bone block (Latarjet procedure): 74 cases]. AB - PURPOSE OF THE STUDY: Results of the Latarjet procedure for chronic anterior shoulder instability using a coracoid block are known in terms of recurrence, but not in terms of apprehension. We studied a prospective consecutive series of shoulders treated with the Latarjet method in order to determine whether patients forget their shoulder or whether they are still bothered, particularly during sports activities. MATERIAL AND METHODS: The series included 74 patients with chronic anterior shoulder instability treated with a coracoid block according to the Latarjet procedure. Sixty-nine were available for review and 66 had complete radiographic explorations (93.2%). The Duplay score was used to classify sports activities. Mean age was 26.5 years; 90% of the patients practiced sports. The surgical procedure was performed by the same operator for 78% of patients. The Duplay score and the Constant score were determined and standard x-rays (four views) were obtained. Statistical analysis was performed with the chi-square test. Multivariate analysis was then applied to the subpopulation presenting persistent apprehension. RESULTS: Follow-up was at least 24 months, average follow-up 50 months. Four patients presented secondary dislocation and two subluxation; 85% of patients were satisfied, 6% were hesitant and 9% were dissatisfied. The Duplay score was: excellent (18.8%), good (49.9%), fair (20.2%) and poor (10.1%). External elbow rotation (RE1) was limited by 17.69 degrees compared with the other side. Eighty-seven percent of patients resumed their sports activity five months postoperatively on average; 34% presented persistent apprehension. This subpopulation was examined separately. Multivariate analysis demonstrated two significantly independent factors of persistent apprehension: recovery of RE1 at 30 days postop, and total recovery of R1 at last follow-up. The radiographies demonstrated degenerative lesions in 10.6% of patients. DISCUSSION: The satisfaction rate of 85% and the 9% failure rate are similar to earlier reports. Our series was however exceptional in terms of an unusually high rate of persistent apprehension. Statistical analysis did not demonstrate a significant link between the presence of hyperlaxity and persistent apprehension. Nevertheless, in patients with hyperlaxity associated with chronic anterior shoulder instability, we associate Latarjet coracoid block with Neer capsuloplasty. PMID- 17401286 TI - [Reverse prosthesis and os acromiale or acromion stress fracture]. AB - PURPOSE OF THE STUDY: We reviewed cases of acromion stress fracture or os acromiale among a consecutive series of 240 reverse prostheses implanted between 1995 and 2003. MATERIAL AND PATIENTS: Ten patients presented os acromiale, ten fracture-fragmentation of the acromion, and one nonunion of the scapular spine. The reverse prosthesis was implanted via a deltopectoral approach with no specific procedure on the acromion. There were 17 women and four men, mean age 72 years. The dominant side was involved for 70%. The prosthesis was indicated for excentered glenoid deterioration (n=14), massive rotator cuff tears without osteoarthritis (n=5), and failure of a hemiarthroplasty (n=2). Seventeen patients were reviewed with more than two years follow-up. RESULTS: 87.5% of the acromial lesions became dislocated postoperatively due to the traction of the deltoid. There was no significant difference between os acromiale and acromion stress fracture. The acromial displacement did not compromise the objective outcome since the Constant score was comparable to that obtained in 179 cases with no acromial lesion reviewed with the same follow-up. Unlike these later shoulders, there were two cases of scapular spine fracture postoperatively, leading to poor function (pain and motion scores less than the global score). CONCLUSION: Acquired or congenital acromial lesions are not a contraindication for reverse prosthesis. Postoperative fracture of the scapular spine is less well tolerated and may required revision for osteosynthesis. PMID- 17401287 TI - [Intra- and interobserver reproducibility of the French version of the Constant Murley shoulder assessment during rehabilitation after rotator cuff surgery]. AB - PURPOSE OF THE STUDY: The Constant-Murley scapular score is currently considered to be the gold standard for shoulder assessment in Europe. Few studies have examined the metrological qualities of this score. Our aim was to study the reliability and validity of the French version in a population undergoing reeducation after rotator cuff surgery. We wanted to determine how pertinent the score is during the reeducation phase (1-12 months after surgery). MATERIAL AND METHODS: Fifty-three patients volunteered to participate in this study. Shoulder assessment was performed by three observers. Intraobserver reproducibility was determined for 102 tests and two series of 32 and 56 tests were used to determine interobserver reproducibility. The internal coherence was studied on a sample of 61 tests. Three observers analyzed the apparent validity of the Constant Murley score. RESULTS: The correlations were satisfactory (intraobserver 0.96; interobserver 0.91 and 0.89 with the Spearman test) and sensitive (intraobserver 0.01; interobserver 0.07 and 0.01 with the Wilcoxon test). Despite satisfactory internal coherence (Cronbach alpha=0.75), the reproducibility of the overall score did not correspond necessarily to the reproducibility of the constituent scores. The measurement error might be related to patient- and observer-related interpretation variability. The apparent validity of the French version might be criticized for assessing rotator cuffs after surgery. CONCLUSION: A precise consensual protocol is needed for conducting the shoulder assessment and establishing the Constant-Murley score during the reeducation phase after rotator cuff surgery. PMID- 17401288 TI - [Early short-term treatment of neonatal hip instability with the Pavlik harness]. AB - PURPOSE OF THE STUDY: The purpose of this work was to study early short-term treatment of reducible dislocated or very unstable dislocated hips using the Pavlik harness. This approach is based on the theory of that dislocation favors dysplasia and that spontaneous correction occurs if the hip is reduced and stable. Very easily dislocated hips have been qualified as "major instability" if voluntary dislocation was easily achieved. In this case, it is very difficult to predict whether postnatal capsuloligamentary retraction will occur in the reduced or dislocated position. MATERIAL AND METHODS: We retained for study 34 patients with 44 pathological hips; 15 hips presented reducible dislocation and 29 major instability. Treatment began early at 3.4 days of life on average and lasted 27.9 days. RESULTS: Outcome was very good for 39 hips with stabilization and spontaneous correction of the acetabular dysplasia, good in four with late correction of the dysplasia, and a failure in one. There were no major complications, none of the patients presented post-reduction osteochondritis. DISCUSSION: Two different entities can be distinguished: dislocation producing dysplasia which is symptomatic neonatally and would imply pathogenic in utero events, and primary dysplasia which can be asymptomatic at birth and for which correction is not spontaneous. If the hip is symptomatic at birth, treatment should begin immediately and be continued to confirmation that the reduced hip is stable, i.e. about three to five weeks on average. Simple dysplasia of dislocatable hips should be excluded from this strategy because of their favorable spontaneous course. Likewise for irreducible dislocated hips because of the higher risk of complications. Under these conditions, this therapeutic strategy is effective for the reduction, stability, and correction of dysplasia achieved in 97.7% of cases without increasing the rate of complications. PMID- 17401289 TI - [Accuracy measurements of acetabular cup positioning using CT less navigation]. AB - PURPOSE OF THE STUDY: Standard methodology is lacking for evaluating the accuracy of surgical navigation systems. The purpose of the present study was to propose a new approach to error measurements of an image-free navigation system used for total hip arthroplasty. MATERIAL AND METHODS: This new approach evaluates the overall accuracy of the system and quantifies the influence of clinical application on this global error. The majority of hip navigation systems use the anterior pelvic plane as part of the reference system. With image-free systems, anatomic pelvic landmarks must be acquired intraoperatively in order to define the anterior pelvic plane. This step could potentially introduce a significant error for navigation. Two studies were performed to measure this error, one on patients and the other on pelvic phantoms. Both used the difference between the intraoperative cup orientation, as displayed by the navigation system and the postoperative cup position, measured on computer tomography (CT) data. The CT measurements used the same reference system as the navigation system. RESULTS: The intraobserver measurement variability ranged from 48.4 degrees to 49.5 degrees for cup abduction and from 12 degrees to 13.5 degrees for anteversion. The interobserver variability ranged from 47.5 degrees to 19 degrees for cup abduction and from 11.8 degrees to 13.8 degrees for anteversion. Overall errors were calculated for cup abduction and anteversion. Cup navigation was accurate on pelvic bone phantoms. The anteversion error ranged from 0 degrees to 2.5 degrees (mean 0.9 degrees, standard deviation 0.7 degrees). For the clinical study, abduction errors ranged from 2.1 degrees to 16.7 degrees. The mean abduction error introduced by the acquisition of anatomic landmarks was 7.2 degrees. DISCUSSION: The proposed simple clinical end-to-end accuracy evaluation model provides the surgeon with sufficiently accurate information. The evaluation model was able to identify and more importantly to quantify the clinically induced error. This study proves that ameliorating the reference system acquisition would improve the system's overall accuracy. PMID- 17401290 TI - [Influence of body mass index on outcome of total hip arthroplasty via a minimally invasive anterior approach]. AB - PURPOSE OF THE STUDY: The purpose of this prospective study was to evaluate the influence of the BMI on the feasibility of minimally invasive total hip arthroplasty (THA). MATERIAL AND METHODS: This prospective study included 86 patients (88 THA) operated on via a single minimally invasive incision (7 cm) using an anterior approach on Judet's orthopedic table. Mean age was 63.7 years and mean BMI was 58.8. Forty-one patients were classified in group I (BMI<25) and 46 in group II (BMI >=25). Perioperative data, pain, postoperative blood loss, duration of the procedure, and length of the incision were noted. All patients were reviewed at three months and radiographic analysis was performed to asses the position of the socket and the quality of the cementation. RESULTS: BMI did not appear to be a strict contraindication for a minimally invasive procedure. Nevertheless, bleeding and operative duration were statistically correlated with BMI (epsilon=4.28 and epsilon=2.66). Extension of the wound noted at the end of the procedure in patients with BMI > =25 (t=5.01) may have resulted from greater pressure on the skin and soft tissue due to stronger traction and more abrasion of the skin edges by reamers and rasps. Such damage may lead to more wound complications even though in our experience there was no statistical difference. On the other hand, socket position and cementation did not appear to be correlated with BMI. DISCUSSION: Results concerning duration of procedure, bleeding, hospital stay and rate of complications are contradictory in the literature. At the same time, criteria for patient selection remain unclear. It thus appeared to be of interest to determine whether BMI was a good criterion to determine the feasibility of THA via a minimally invasive procedure. CONCLUSION: BMI appears to be a good criterion to evaluate the risk of wound complications after minimally invasive surgery. On the other hand, other criteria must be found since BMI does not evaluate muscle mass which seems to be of major concern in minimally invasive procedures. PMID- 17401291 TI - [Clinical and posturographic comparison of patients with recent total hip arthroplasty]. AB - PURPOSE OF THE STUDY: To highlight the congruence of clinical and posturographic tests in patients undergoing hip arthroplasty. MATERIAL AND METHODS: Ten patients (six males and four females) were included in this study and tested when at admission and discharge from the rehabilitation department (12 and 27 days after surgery respectively). The patients were asked to stand undisturbed in the upright position, eyes closed on a system composed of two force platforms. Five successive 32s trials (sampled at 64 Hz) were conducted with rest intervals of similar duration between trials. The plantar center of pressure (CP), displacements, and resultant CP (CPRes) were then computed and analyzed in various ways. In parallel, various clinical tests, including muscular force, hip range of motion, walking speed, functional independence, pain, sensitivity, lateral reach, and get up and go aimed at evaluating global coordination. Correlations, using the non-parametric Spearman coefficient, were computed from the differences between clinical and posturographic parameters obtained at admission and discharge from the rehabilitation department. RESULTS: Certain statistically significant differences in postural behavior was observed both for clinical and posturographic tests between admission and discharge. Body weigh distribution over the two legs was largely asymmetric at onset and, though it declined, persisted at discharge. There was not difference for the mean positions of both the plantar CP and the resultant of the CP movements. On the contrary, it was noteworthy that the variances of CP displacements (data dispersion) were initially larger on the sound leg along the anteroposterior axis and that this compensatory feature disappeared at the end of the stay. At discharge, the variances computed from the sound and the prosthetic limb became equivalent. All the clinical tests demonstrated statistically significant improvements in results at discharge compared with admission. Several significant correlations involving clinical tests and mean positions along the anteroposterior axis, the degree of body weight asymmetry and variances along both the mediolateral and anteroposterior axes were found. DISCUSSION: These results enable a better understanding of strategies used by hip arthroplasty patients to keep balance. Even though their sensitivity was quite different, both clinical and posturographic measurements demonstrated their ability to assess recovery from surgery. These two evaluation techniques are complementary. PMID- 17401292 TI - [Superior mesenteric artery syndrome following correction of spinal deformity: case report and review of the literature]. AB - Superior mesenteric artery syndrome is a rare complication which can develop after surgical correction of a spinal deformity. The syndrome is caused by an extrinsic compression on the third portion of the duodenum by the aorta posteriorly and the mesenteric artery anteriorly. We report here a case of aortomesenteric compression of the duodenum secondary to surgical correction of lower thoracic scoliosis in a 19-year-old female. The patient presented vomiting and intestinal obstruction ten days after spinal surgery. Treatment consisted in exclusive parenteral nutrition followed by careful surveillance and progressive reintroduction of oral food intake to avoid unnecessary surgery. Young thin subjects are predominantly exposed to this type of complication. The body mass index is a good indication to identify subjects at risk. Symptoms of upper gastrointestinal obstruction develop seven to ten days after surgery. Diagnosis is based on transit studies using a hydroluble contrast agent which reveals major gastric dilation and a clear interruption of the transit at the level of the third duodenum as well as retrograde peristaltism. Medical treatment should be undertaken first and is effective in the large majority of cases. Surgery may be proposed only in the event of failure. Recurrence is exceptional. Early diagnosis, delivery of clear information for the patient and family and multidisciplinary management are important points to consider for proper care for this complication which if neglected can become life-threatening. PMID- 17401293 TI - [A pediatric case of rupture of the brachial artery and radial nerve palsy secondary to proximal humeral exostosis]. AB - Rupture of the brachial artery associated with radial nerve palsy in a context of exostosis of the proximal humerus has not been described to date in the literature. Our patient was a fourteen-year-old girl with a history of violent pain occurring suddenly with no prodrome or triggering factor. The pain was localized at the level of the proximal left humerus. Physical examination revealed the presence of a hematoma and complete motor radial nerve palsy. The diagnosis was not confirmed by computed tomography with contrast injection but was confirmed by magnetic resonance imaging which eliminated malignant transformation of the exostosis. After checking the neurovascular bundles and evacuating the hematoma, treatment consisted in resection of the exostosis and arterial repair with an autologous venous graft. We discuss the diagnostic and therapeutic challenges which present vascular complications due to exostosis. PMID- 17401294 TI - [Elbow dislocation in children associated with proximal radioulnar translocation]. AB - We report a unique case of posterior elbow dislocation with proximal radioulnar translocation in a six-year-old child with 3D reconstruction CT images. Five other cases of this type of association have been reported in the literature. Diagnosis is often established late. Our case illustrates the diagnostic and therapeutic difficulties. The CT scan demonstrated the presence of the proximal radioulnar translocation and contributed greatly to understanding the pathogenic mechanism: the radius crossed over the ulna anteriorly. The CT-scan can reveal associated lesions missed on the plain x-rays. Surgical reduction is generally needed, preferably via a medial approach. PMID- 17401295 TI - [Chondroblastoma in children. A SOFOP series (French Society of Pediatric Orthopedics): 89 cases]. PMID- 17401296 TI - HDL function as a target of lipid-modifying therapy. AB - High-density lipoprotein (HDL) is conventionally believed to possess many features that protect against atherosclerosis. However, these lipoproteins may be modified in certain individuals and/or circumstances to become pro-inflammatory. The ability of HDL to inhibit or paradoxically to enhance vascular inflammation, lipid oxidation, plaque growth, and thrombosis reflects changes in specific enzyme and protein components. The anti-inflammatory and pro-inflammatory functional properties of HDL can now be assessed using cell-based and cell-free assays. Acute or chronic systemic inflammation and the metabolic syndrome appear to render HDL pro-inflammatory. In contrast, statins and experimental agents such as apolipoprotein A-1 mimetics render HDL more anti-inflammatory. The 2 main classes of existing drugs for HDL modification are fibric acid derivatives, also known as "fibrates," and niacin- containing compounds. In several controlled and prospective intervention studies, patients with low HDL-C and additional risk factors benefited from treatment with fibrates or niacin. However, an increase in HDL-C did not lead to a decrease in cardiovascular events in all trials. PMID- 17401297 TI - The relationship among risk factor clustering, abdominal obesity, and residual risk for cardiovascular events. AB - Statins, angiotensin-converting enzyme inhibitors, and combination therapies have been shown to reduce the cardiovascular event rate in susceptible individuals, albeit with remaining significant residual risk. Some of the sources of residual risk, such as genetics and epigenetic phenomena, are not easily modifiable. Still, the risk imposed by these factors may be lowered by implementation of dietary, behavioral, and pharmacologic interventions. Abdominal obesity has emerged as one element in the cluster of factors linked to increased propensity for cardiovascular disease and type 2 diabetes. It is a potential therapeutic target to reduce residual cardiometabolic risk. Waist circumference has been shown to be a strong correlate of abdominal obesity, and measurement is a useful tool for the assessment of cardiometabolic risk. PMID- 17401298 TI - Noninvasive coronary angiography using multislice computerized tomography. AB - Multislice computerized tomography (MSCT) is a relatively new, noninvasive method for evaluating coronary stenosis. In symptomatic patients, the use of MSCT has been shown to be effective in identifying coronary lesions with comparable accuracy to the traditional, catheter-based invasive angiography. With the 64 slice MSCT scanner, units are acquired faster and the slices are thinner, resulting in improved temporal and spatial resolution and coronary artery imaging. Additional benefits of the 64-slice MSCT scanner as compared with the older generation scanners include a shorter period during which the patient must hold his or her breath, a wider range of acceptable heart rates, and the ability to image very obese patients and those with moderate coronary calcium with lower volumes of contrast. The 64-slice MSCT scanner has some limitations, but it demonstrates an improved image resolution that allows for a more precise evaluation. This new generation of MSCT scanners may eventually eliminate the need for invasive coronary angiography. PMID- 17401301 TI - Acute myocardial infarction in a patient with systemic lupus erythematosus and normal coronary arteries. AB - Although cardiac manifestations such as pericardial, myocardial, and valvular involvement are common in patients with systemic lupus erythematosus (SLE), coronary artery involvement is less frequent. Clinical manifestations of coronary artery disease in SLE can result from accelerated atherosclerosis, arteritis, abnormal coronary flow reserve, spasm, and thrombosis. In SLE, the classic valvular abnormality consists of noninfective, verrucous vegetation. Thickening of the leaflets due to inflammation followed by fibrosis is common, occurring in about 50% of patients, whereas vegetations are present in about 40%. Mitral valve involvement is most common, with valvular regurgitation more frequent than valvular stenosis. The tricuspid valve and the aortic valve may also be affected. Its frequency varies widely: 13% to 74% in the general population. We report a case of a woman with acute myocardial infarction and normal coronary arteries, who was subsequently diagnosed with Libman-Sacks endocarditis and SLE. PMID- 17401303 TI - Nesiritide for acute decompensated heart failure. PMID- 17401302 TI - Spontaneous coronary artery dissection in a woman on fenfluramine. AB - Spontaneous coronary artery dissection (SCAD) is a rare cause of acute coronary syndrome, cardiogenic shock, and sudden cardiac death in women of reproductive age who have no traditional risk factors for coronary artery disease. The etiology, prognosis, and treatment of SCAD remain poorly defined. Coronary angiography is the gold standard for diagnosis. Management includes medical therapy and revascularization procedures using percutaneous intervention and coronary artery bypass grafting. Possible mechanisms of SCAD include rupture of atherosclerotic plaque or vasa vasorum, hemorrhage between the outer media and external lamina with intramedial hematoma expansion, and compression of the vessel lumen. We report a case of SCAD in a 39-year-old woman presenting with ST elevation myocardial infarction midway through her menstrual cycle. Her medications included fenfluramine for obesity and hydrochlorothiazide, amlodipine, and atenolol for hypertension. PMID- 17401304 TI - Estimated glomerular filtration rate: why is it important? PMID- 17401305 TI - Clinical challenges in the prevention and treatment of late and very late stent thrombosis. Introduction. PMID- 17401306 TI - Incidence of late stent thrombosis with bare-metal, sirolimus, and paclitaxel stents. AB - Stent thrombosis has become a major concern for interventional cardiology. Although infrequent, it is associated with significant morbidity and mortality. Recent attention has focused on the frequency of this complication with drug eluting stents compared with bare-metal stents in regard to the timing (early, late, or very late) of the event, underlying mechanisms involved, and preventive strategies. Although dual antiplatelet therapy (aspirin plus thienopyridine) is crucial in mitigating the problem, there are significant issues with this management strategy, including the duration of dual antiplatelet treatment, patient compliance, variability in individual response to therapy, bleeding risk, and management of subsequent noncardiac surgical procedures. Newer strategies being evaluated to enhance the safety of drug-eluting stents include different alloys and stent designs, revisions in the polymer or drug utilized, and, ultimately, bioabsorbable platforms. PMID- 17401307 TI - Stent thrombosis: role of compliance and nonresponsiveness to antiplatelet therapy. AB - Percutaneous coronary intervention with drug-eluting stents has revolutionized the management of patients with symptomatic coronary artery disease. Although this strategy significantly reduces the incidence of restenosis and repeat revascularization, concern has been raised about an increased frequency of late stent thrombosis with drug-eluting stents compared with bare-metal stents. The mechanism of stent thrombosis remains unclear, and various hypotheses have been described. Platelets are believed to play a pivotal role in the development of stent thrombosis, with pathological studies demonstrating an abundance of platelets within the occlusive thrombi. Premature discontinuation and nonadherence to antiplatelet therapy are considered important risk factors for late stent thrombosis. Early identification of vulnerable patients and definition of the role of antiplatelet nonresponsiveness in the development of stent thrombosis should be the focus of future diagnostic and therapeutic strategies. PMID- 17401308 TI - Primary prevention, treatment, and secondary prevention of late and very late stent thrombosis. AB - The occurrence of late and very late thrombotic complications in association with drug-eluting stents (DES) has recently garnered much attention, but these complications are also associated with bare-metal stents (BMS). Predisposing factors for BMS thrombosis, both early and late, include noncompliance with antiplatelet agents, an exercise-induced procoagulant state, brachytherapy, small stent size, underdeployment of the stent, depressed left ventricular ejection fraction, and impaired response to antiplatelet therapy. Independent predictors of DES thrombosis include premature interruption of antiplatelet therapy, primary stenting in acute myocardial infarction, total stent length, and renal failure. Prevention depends on applying an optimal stent deployment technique at the time of the index percutaneous coronary intervention (PCI), compliance with dual antiplatelet therapy, and extending antiplatelet therapy beyond current package insert recommendations. In patients who develop late stent thrombosis, efforts to achieve rapid normalization of coronary blood flow with PCI are mandatory. PMID- 17401309 TI - Neointimal formation following drug-eluting stents: physiology, timeline, and the influence of drug delivery systems. AB - Percutaneous coronary intervention with drug-eluting stents (DES) is currently the preferred approach to the treatment of obstructive coronary artery disease. Large, randomized trials have demonstrated a significant reduction in the incidence of restenosis and the need for target vessel revascularization following implantation of DES compared with bare-metal stents. Follow-up data extending out to 2 to 4 years have demonstrated efficacy in maintaining luminal patency, but recent concerns regarding potential late adverse effects with DES have been raised. These include aneurysm formation and hypersensitivity reactions, as well as subacute and late stent thrombosis requiring compliance with antiplatelet therapy for protracted periods of time. Evolving strategies to mitigate late adverse events with DES include acceleration of endothelialization, gene therapy targeting pro-healing pathways (ie, nitric oxide donors), smooth muscle cell growth inhibitors, bioabsorbable metal and polymeric stents, and concurrent use of local as well as systemic chemotherapy. PMID- 17401310 TI - New drug-eluting stent platforms to prevent stent thrombosis. AB - Although earlier reports from randomized controlled clinical trials suggested that the incidence of stent thrombosis following drug-eluting stent (DES) implantation was similar to or less than that observed following bare-metal stent deployment, longer-term follow-up has revealed a persistent, protracted risk for thrombosis following DES. This apparent divergence in risk for thrombosis becomes evident beyond 6 to 12 months following deployment. The proposed etiologies of late DES thrombosis are multifactorial and differ somewhat from those factors incriminated in bare-metal stent thrombosis. Prevention strategies are in development to address polymer hypersensitivity/inflammatory response, delayed endothelialization/vessel healing, late incomplete stent apposition, persistence of the underlying endoluminal metal prosthesis, and discontinuation of antiplatelet therapies. PMID- 17401311 TI - Expanding the opportunities for blocking the renin-angiotensin system: introduction to a special supplement. AB - The previous dogma that the renin-angiotensin system exerts its effects entirely through angiotensin II is now under challenge as scientists explore the properties of the prorenin/renin receptor and start to study local vascular actions of renin independent of its production of angiotensin in the plasma. The demonstrated blood pressure effects of the first clinically developed renin inhibitor, aliskiren, have confirmed the validity of this new class of drugs. Future research, exploring effects on the renin-angiotensin system that perhaps cannot be provided by the currently used blockers of this system, will test whether enhanced clinical benefits might result from this new pharmacologic strategy in patients at risk of cardiovascular events. PMID- 17401312 TI - Renin-angiotensin system modulation for treatment and prevention of cardiovascular diseases: toward an optimal therapeutic strategy. AB - The unraveling of the role of the renin-angiotensin system (RAS) in health and disease is an example of how basic and applied scientists can decipher a complex biological system to better understand the pathophysiology of disease. Moreover, clinicians have been provided with drugs to modulate the RAS, including the angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs). ACE inhibitors and ARBs have revolutionized the way in which many diseases are treated, including hypertension, heart failure, diabetes mellitus, and kidney disease. Yet, despite the undoubted successes of these drugs, cardiovascular morbidity and mortality remain high. Clearly, lower blood pressure goals may be required. Because ACE inhibitors and ARBs target specific areas of the RAS, more impressive results might be obtained with a more global reduction in RAS activity. This article examines the results of clinical trials of ACE inhibitors and ARBs and assesses the potential for improving outcomes through a more global inhibition of the RAS with renin inhibitors. PMID- 17401313 TI - The efficacy of aliskiren, a direct renin inhibitor, in the treatment of hypertension. AB - Aliskiren is a potent, highly specific renin inhibitor with better oral bioavailability than earlier renin inhibitors and a long plasma half-life that makes it suitable for once-daily dosing. The efficacy and safety of aliskiren in treating hypertension has been studied in clinical trials both as monotherapy, comparing it with existing antihypertensive therapies, and in combination with other antihypertensive agents, including the diuretic hydrochlorothiazide, the angiotensin-converting enzyme inhibitor ramipril, and the calcium channel blocker amlodipine. From the extensive database acquired to date, it is clear that aliskiren is an effective antihypertensive agent, with once-daily administration resulting in dose-dependent systolic and diastolic blood pressure reductions. Combinations with existing antihypertensives are producing promising additional blood pressure-lowering effects. PMID- 17401314 TI - The renin system: is direct renin inhibition different from blockade at the AT1 receptor or the ACE step? AB - A substantial level of evidence supports the use of renin system blockade for many patients with hypertension. Two lines of evidence, based on very high-dose angiotensin blocker treatment or combination therapy with angiotensin-converting enzyme inhibitor and angiotensin receptor blocker, suggest that more complete blockade leads to improved clinical outcomes. The recent development of a powerful renin inhibitor that acts at the initial, rate-limiting step in the renin cascade would also favor more complete blockade of the system. For many patients, this is likely to lead to improved treatment. PMID- 17401315 TI - Intracameral 0.5% phenylephrine--a safe solution? PMID- 17401316 TI - The difference in muscle slippage according to scleral suture techniques in rectus muscle resection of rabbit eyes. AB - PURPOSES: To investigate the degree of muscle slippage according to scleral suture techniques in extraocular muscle resection of rabbit eyes. METHODS: Twenty eyes from 10 rabbits (20 superior rectus muscles) were divided into four groups according to scleral suture techniques. The types of scleral suture technique used for the four groups were as follows: group 1--double-arm mattress suture technique without midline suture; group 2--double-arm mattress suture technique with midline suture; group 3--long scleral tunnel suture technique without midline suture; and group 4--long scleral tunnel suture technique with midline suture. Five superior rectus muscles were assigned to each group and they underwent resection with 6-0 prolene using one of the four suture techniques. The degree of muscle slippage was measured 3 months after the surgery, defined as the distance between the prolene materials at the centre of muscle insertion and the pre-placement suture. RESULTS: The mean degree of muscle slippage for treatment groups 1-4 was 2.3+/-0.62, 1.0+/-0.27, 1.5+/-0.45, 0.5+/-0.46 mm respectively. Group 1 had significantly more muscle slippage than groups 2-4 (P=0.006, P=0.046, P=0.001 respectively). Group 4 had the least slippage among the four groups, while group 3 had significantly more slippage than group 4 (P=0.009). The differences between groups 2 and 3 and between groups 2 and 4 were not statistically significant (P=0.083, P=0.077 respectively). CONCLUSION: Long scleral suture technique is more effective method than double-arm mattress suture technique for the prevention of muscle slippage in rectus muscle resection. The addition of a midline suture of rectus muscle was helpful in the prevention of muscle slippage. PMID- 17401317 TI - Peripheral ulcerative keratitis in pityriasis rubra pilaris. PMID- 17401318 TI - Orbital cellulitis following intralesional corticosteroid injection for periocular capillary haemangioma. PMID- 17401319 TI - Retinal nerve fibre layer characteristics with vigabatrin-associated visual field loss--could scanning laser polarimetry aid diagnosis? AB - PURPOSE: To describe the retinal nerve fibre layer (RNFL) characteristics in patients suffering vigabatrin-associated visual field loss or behaviour consistent with field loss. METHODS: Eight patients with visual field loss attributed to vigabatrin use had their RNFL evaluated by scanning laser polarimetry. RESULTS: All eight patients managed to perform the test reliably. The mean length of vigabatrin treatment was 81 months with a mean cumulative dose of vigabatrin being 5.4 kg. All patients had significantly reduced RNFL parameters; mean TSNIT=36.5 microm, mean nerve fibre indicator=63.1, mean superior average=42.7 microm, and mean inferior average=39.2 microm. There was no correlation between cumulative dose or time on treatment and RNFL thickness. CONCLUSION: Scanning laser polarimetry can be performed safely and reliably on patients with vigabatrin-associated field loss. RNFL thickness is reduced in these patients with vigabatrin-associated field loss. RNFL loss concentrates on the long nerve fibres, which explains the nasal pattern of field loss seen in these patients. This investigation shows promise in the screening of vigabatrin patients, especially in those patients that may not tolerate formal field testing well. PMID- 17401320 TI - Emulsification of Densiron-68 used in inferior retinal detachment surgery. AB - PURPOSE: To report the clinical features of eight patients presenting with emulsification of the heavier than water vitreous substitute, Densiron-68. METHODS: Two patients underwent primary inferior retinal detachment (RD) surgery, two patients underwent giant retinal tear repair, three patients had repair of inferior RD complicated by proliferative vitreoretinopathy and one patient had inferior RD surgery following repair of a scleral rupture. All patients had insertion of Densiron-68. RESULTS: Significant emulsification of Densiron-68 was seen within 12 weeks of surgery in eight cases out of a total of 40 patients receiving Densiron-68. CONCLUSION: Despite adequate Densiron fills, emulsification necessitated its removal. Emulsified Densiron may have contributed to significant intraocular inflammation, epiretinal membrane formation and cystoid macular oedema. Without removal, prolonged presence of emulsified Densiron may lead to keratopathy, secondary glaucoma and retinal toxicity secondary to partitioning of perfluorohexyloctane. This has potentially significant implications on the indications for Densiron-68 use and warrants consideration before deciding on the optimal surgical intervention for inferior RDs. PMID- 17401321 TI - Macular pigment in the human retina: histological evaluation of localization and distribution. AB - PURPOSE: Clinical investigations have demonstrated variation in both the peak optical density and the spatial distribution of macular pigment. To confirm these impressions histologically, the present study examined the distribution of macular pigment in the human retina. MATERIALS AND METHODS: The macular retina of 11 donor eyes of different ages (28-91 years) were examined histologically on 100 microm vibratome sections directly, without further staining. Measurements were made in two dimensions: (1) adding the number of macular sections with visible macular pigment, and (2) direct measurement of the extension of macular pigment in the foveolar section, which visibly contained the most macular pigment. RESULTS: The measurements with two methods demonstrated good correlation. The macula demonstrated a variation in the spatial extension of the visible macular pigment between 200 and 900 microm diameter around the centre of the fovea, which was also found when direct measurements were taken. There was no correlation with the donor age. The main location of macular pigment was in the layer of the fibres of Henle in the fovea and in the inner nuclear layer at the parafoveal site. CONCLUSIONS: Histologically, a wide variation of the spatial distribution of macular pigment was found that confirms clinical observations. The primary localization of human macular pigment is in the inner retinal layers. PMID- 17401322 TI - Ocular disorders in a petroleum industry in Nigeria. AB - OBJECTIVE: To determine the ocular disorders in the petroleum industry in Warri, Nigeria. METHODS: A clinic-based prospective study of the pattern of ocular disorders in workers of the Nigerian National Petroleum Corporation in Warri, Delta state of Nigeria was carried out over a 1-year period. Technical and non technical workers were interviewed and examined using the Snellen's chart, pen torch, direct ophthalmoscope, slit-lamp biomicroscope, Perkin's hand-held applanation tonometer, and the Kowa automatic visual field plotter where indicated. RESULTS: A total of 154 technical workers comprising 146 males (94.8%) and eight females (5.2%), and 116 non-technical workers comprising 90 males (77.6%) and 26 females (22.4%) were seen. There was no reported case of ocular trauma. Symptoms suggestive of ocular irritation were much more common in technical workers. Allergic conjunctivitis, pterygium, corneal abrasion, and foreign body on the cornea were significantly more common in technical workers P<0.0001. CONCLUSIONS: Exposure to irritant chemicals in the petroleum industry makes the technical workers more prone to allergic conjunctivitis, pterygium, corneal abrasion, and foreign body on the cornea. Protection goggles should be provided for all the technical workers. PMID- 17401323 TI - NEMO mutational analysis in a Japanese family with incontinentia pigmenti. PMID- 17401324 TI - Surgical outcomes for vitrectomy in Eales' disease. AB - AIM: To present and analyse the anatomical and functional outcomes for vitrectomy in Eales' disease. METHODS: This retrospective case series enrolled 63 patients (71 eyes) who underwent pars plana vitrectomy (PPV) for the complications of Eales' disease. Indications included nonclearing vitreous haemorrhage (VH) with/without epiretinal membranes in 49 (69%) eyes, and retinal detachments (RD) involving/threatening macula in 22 (31%). Additional procedures (scleral buckling; gas/oil tamponade) were performed at surgeon's discretion. Minimum follow-up was 6 months. The primary outcome measures were change in best corrected visual acuity (BCVA) and posterior segment status. RESULTS: The mean age of the patients was 29.6 years (range: 15-70 years); 60 were male (95%). Preoperatively, posterior vitreous detachment (PVD) existed in 28 (39.4%) eyes. Forty (56.3%) eyes underwent only PPV; the rest required additional surgical procedures (q.v.). The mean baseline BCVA (1/60) improved to 6/24 postoperatively (P<0.0001). Fifty-four eyes of 50 patients (76%) showed an improvement of > or =2 equivalent Snellen lines; six eyes (four patients) remained stable (+/-1 line); visual acuity worsened in 11 eyes (nine patients). The mean final BCVA was similar in eyes operated for VH and RD (P=0.08); but the magnitude of change from baseline was greater in the VH group (P=0.009). PVD had a borderline association with final BCVA (P=0.056); but did not influence the functional/anatomical improvement. Thirteen eyes required repeat interventions; 11 (15.49%) eyes experienced surgical failure. CONCLUSIONS: Although surgical outcomes in Eales' disease depend on preoperative PVD/RD to some extent; good results are possible in the presence of incomplete PVD and tractional sequelae. PMID- 17401325 TI - Full macular translocation following photodynamic therapy in neovascular age related macular degeneration. AB - PURPOSE: To report the long-term functional and anatomical outcome of full macular translocation (FMT) in eyes with neovascular age-related macular degeneration (AMD) following photodynamic therapy (PDT). METHODS: Twelve eyes of 12 consecutive patients with neovascular AMD who were PDT-nonresponders and underwent FMT were analysed. Best-corrected visual acuity (BCVA) measurement, fundus photography, and fluorescein angiography at baseline and at follow-up examinations in 3 months intervals were performed. Primary end point was change of BCVA from baseline to last visit. RESULTS: Totally 12 eyes of 12 patients were analysed. Mean time interval between the last PDT and FMT was 3.7 months (range 1 10 months). Mean follow-up after FMT was 25.6 months. BCVA ranged at baseline from 20/1000 to 20/80 (mean 20/230). At the last visit, mean BCVA was by 20/185. BCVA improved in 50% (6/12) of eyes by more than 1 line. Twenty five per cent (3/12) of eyes had final BCVA within +/-1 line from baseline. In 25% (3/12) of eyes the BCVA decreased by more than 1 line. One eye had recurrent CNV. In four eyes a cystoid macular oedema developed. No retinal detachment or disturbing diplopia was noted. CONCLUSIONS: In the present study, FMT in PDT-nonresponders stabilised or improved visual acuity in the majority of the eyes in a mean follow up period of nearly 2 years. FMT can be considered as a therapeutical option in eyes who are nonresponders to the PDT in neovascular AMD. PMID- 17401326 TI - The new malononitrilamide immunosuppressant FK778 prolongs corneal allograft survival in the rat keratoplasty model. AB - PURPOSE: Aim of this study was to prove the efficacy and safety of the new malononitrilamide immunosuppressive FK778 in prolonging clear graft survival following allogeneic orthotopic keratoplasty in rats. METHODS: Sixty-seven penetrating keratoplasties were performed using Fisher and Lewis rats as donors and recipients, respectively: group 1 (n=11), allogeneic control without therapy; group 2 (n=12), syngeneic control; group 3 (n=11), mycophenolate mofetil (MMF) 40 mg/kg bodyweight; group 4 (n=12), FK778 5 mg/kg bodyweight; group 5 (n=12), FK778 10 mg/kg bodyweight; and group 6 (n=9), FK778 20 mg/kg bodyweight. Four animals in each group were killed for immunohistological evaluation on day 14. Therapy was administered orally for 18 days. The grafts were evaluated every three days by means of a scoring system including opacity, oedema, and vascularization. Time to rejection was analysed with the Kaplan-Meier survival analysis and compared with the log-rank test. The densities of infiltrating immune cells were compared statistically using the non-parametric Mann-Whitney test. RESULTS: Mean rejection free graft survival was 11.4 days in group 1 (allogeneic control), 100 days (total follow-up time) in group 2 (syngeneic control), 24.0 days in group 3 (MMF 40 mg/kg), 15.7 days in group 4 (FK778 5 mg/kg), 19.1 days in group 5 (FK778 10 mg/kg), and 25.4 days in group 6 (FK778 20 mg/kg) (P<0.005). CONCLUSIONS: Systemic immunosuppression with FK778 prolongs graft survival in the rat keratoplasty model. FK778's efficacy is comparable with that of MMF in preventing immunologic graft rejection. PMID- 17401327 TI - Non-Hodgkin's lymphoma presenting with nystagmus: an unusual case. PMID- 17401328 TI - 3D vs 2D qualitative and semiquantitative evaluation of the glaucomatous optic disc atrophy using computer-assisted stereophotography. AB - BACKGROUND AND PURPOSE: Real colour documentation of the optic nerve head (ONH) is one of the most important methods to identify the early progression of glaucomatous optic nerve damage. This study compares the qualitative and semiquantitative evaluation of ONH photographs, using a 3D time-multiplexing system and conventional 2D photography, visualised on a computer monitor. PATIENTS AND METHODS: Stereophotographs of the ONH were converted by special software in a 3D file. The same images were shown in 2D and 3D to 22 subjects with different training grades (residents and ophthalmologists). The evaluation based on a standardised questionnaire, containing semiquantitative parameters of the ONH. A weighted score for quality of ONH assessment counted the correct answers of the observers to compare their evaluation in 2D to the one made in 3D. RESULTS: All of the observers achieved a significant better evaluation of the ONH with 3D than with 2D images. No significant differences were observed between both groups. Classification errors with respect to the 'gold standard' were lower for 3D measurements as compared with 2D measurements (P=0.007). No significant differences were observed between the gold standard and residents or ophthalmologists for the ONH size, but for the c/dratio the stereoscopic images lead to better results. CONCLUSION: This study showed a better evaluation of parameters associated with the glaucomatous ONH atrophy with 3D images compared with 2D, independent of the clinical training grade. The computer-based evaluation of the ONH atrophy by using a time-multiplexing system (shutter glasses) may improve the diagnosis of glaucoma patients. PMID- 17401329 TI - Live-cell assay for detection of apoptosis by dual-laser flow cytometry using Hoechst 33342 and 7-amino-actinomycin D. AB - This protocol describes a rapid and simple method for the identification of apoptotic cells. Owing to changes in membrane permeability, early apoptotic cells show an increased uptake of the vital DNA dye Hoechst 33342 (HO342) compared with live cells. The nonvital DNA dye 7-amino-actinomycin D (7-AAD) is added to distinguish late apoptotic or necrotic cells that have lost membrane integrity from early apoptotic cells that still have intact membranes as assayed by dye exclusion. The method is suitable to be combined with cell surface staining using Abs of interest labeled with fluorochromes that are compatible with HO342 and 7 AAD emissions. Surface antigen staining is carried out according to standard methods before staining for apoptosis. The basic assay can be completed in 30 min, and extra time is needed for cell surface antigen staining. PMID- 17401330 TI - Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method. AB - Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially useful for laboratories using one or two strains repeatedly. The production of yeast cells for freezing takes only approximately 30 min, once the yeast culture has grown up. Transformation with frozen competent yeast cells will take approximately 30 min, depending on the heat shock used. PMID- 17401331 TI - Microtiter plate transformation using the LiAc/SS carrier DNA/PEG method. AB - Here, we describe a protocol that has been adapted for the transformation of yeast cells in 96-well microtiter plates. This protocol can be tailored for multiple applications and is suitable for high-throughput applications. It can be completed in 2-3 h, once the yeast cells have been grown depending on the heat shock used. PMID- 17401332 TI - Serial sectioning and electron microscopy of large tissue volumes for 3D analysis and reconstruction: a case study of the calyx of Held. AB - Serial section electron microscopy is typically applied to investigation of small tissue volumes encompassing subcellular structures. However, in neurobiology, the need to relate subcellular structure to organization of neural circuits can require investigation of large tissue volumes at ultrastructural resolution. Analysis of ultrastructure and three-dimensional reconstruction of even one to a few cells is time consuming, and still does not generate the necessary numbers of observations to form well-grounded insights into biological principles. We describe an assemblage of existing computer-based methods and strategies for graphical analysis of large photographic montages to accomplish the study of multiple neurons through large tissue volumes. Sample preparation, data collection and subsequent analyses can be completed within 3-4 months. These methods generate extremely large data sets that can be mined in future studies of nervous system organization. PMID- 17401333 TI - A convenient, high-throughput assay for measuring the relative cell permeability of synthetic compounds. AB - We describe here a convenient procedure for assessing the relative cell permeability of chemical compounds. The assay can be used in a high-throughput mode and is particularly applicable for the evaluation of the relative permeability of compounds in a combinatorial library. The compound of interest is conjugated to a dexamethasone derivative. The entry of the conjugate into living mammalian cells triggers the nuclear transport of a Gal4 DNA binding domain glucocorticoid receptor ligand binding domain-VP16 activation domain fusion protein and, consequently, the activation of a Gal4-responsive luciferase reporter gene. The relative cell permeability is thus described quantitatively by the level of luciferase expression. The experiments take only a few days once chemical synthesis and conjugation are finished. PMID- 17401334 TI - High-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. AB - Here we describe a high-efficiency version of the lithium acetate/single-stranded carrier DNA/PEG method of transformation of Saccharomyces cerevisiae. This method currently gives the highest efficiency and yield of transformants, although a faster protocol is available for small number of transformations. The procedure takes up to 1.5 h, depending on the length of heat shock, once the yeast culture has been grown. This method is useful for most transformation requirements. PMID- 17401335 TI - Quick and easy yeast transformation using the LiAc/SS carrier DNA/PEG method. AB - Here, we describe a quick and easy version of the lithium acetate/single-stranded carrier DNA/PEG method of transformation for Saccharomyces cerevisiae. This method can be performed when only a few transformants are needed. The procedure can take less than an hour, depending on the duration of the heat shock. It can be used to transform yeast cells from various stages of growth and storage. Cells can be transformed from freshly grown cells as well as cells stored on a plate at room temperature or in a refrigerator. PMID- 17401336 TI - Large-scale high-efficiency yeast transformation using the LiAc/SS carrier DNA/PEG method. AB - Here, we describe a Library screen transformation protocol using the lithium acetate/single-stranded carrier DNA/PEG method of transformation for Saccharomyces cerevisiae. This method is suitable for screening complex plasmid libraries such as those used for yeast two-hybrid analysis. This procedure takes up to 2.5 h to complete once the yeast culture has been grown. PMID- 17401337 TI - Synthesis of TOAC spin-labeled proteins and reconstitution in lipid membranes. AB - A procedure is described for the synthetic incorporation into membrane proteins of the non-natural amino acid TOAC (2,2,6,6-tetramethyl-piperidine-1-oxyl-4-amino 4-carboxylic acid), which is coupled rigidly to the alpha-carbon, providing direct detection of peptide backbone dynamics by electron paramagnetic resonance (EPR). Also included is a protocol for the functional reconstitution of the spin labeled protein in lipid vesicles. This protocol can be completed in 17 d. PMID- 17401338 TI - Anti-primer quenching-based real-time PCR for simplex or multiplex DNA quantification and single-nucleotide polymorphism genotyping. AB - Nucleic acid amplification and detection plays an increasingly important role in genetic analysis of clinical samples, medical diagnostics and drug discovery. We present a new quantitative PCR method that allows versatile and flexible nucleic acid target quantification. One of the PCR primers is modified by an oligonucleotide "tail" fluorescently labeled at the 5' end. An oligonucleotide complementary to this tail, carrying a 3'-quencher ("anti-primer"), is included in the PCR along with the two primers. Following primer extension, the reaction temperature is lowered such that the anti-primer hybridizes to and quenches the fluorescence of only the free primer and not the double-stranded PCR product, allowing real-time fluorescent quantification of the latter. This anti-primer based quantitative real-time PCR (aQRT-PCR) allows simplex or multiplex quantification or single-nucleotide polymorphism genotyping in clinical samples of widely differing quality (e.g., fresh samples, formalin-fixed paraffin embedded samples and plasma-circulating DNA) and provides a practical alternative to existing, more expensive approaches. The process of aQRT-PCR takes 1.5-2 h. PMID- 17401339 TI - Simultaneous mutation scanning and genotyping by high-resolution DNA melting analysis. AB - This protocol permits the simultaneous mutation scanning and genotyping of PCR products by high-resolution DNA melting analysis. This is achieved using asymmetric PCR performed in the presence of a saturating fluorescent DNA dye and unlabeled oligonucleotide probes. Fluorescent melting curves of both PCR amplicons and amplicon-probe duplexes are analyzed. The shape of the PCR amplicon melting transition reveals the presence of heterozygotes, whereas specific genotyping is enabled by melting of the unlabeled probe-amplicon duplex. Unbiased hierarchal clustering of melting transitions automatically groups different sequence variants; this allows common variants to be easily recognized and genotyped. This technique may be used in both laboratory research and clinical settings to study single-nucleotide polymorphisms and small insertions and deletions, and to diagnose associated genetic disorders. High-resolution melting analysis accomplishes simultaneous gene scanning and mutation genotyping in a fraction of the time required when using traditional methods, while maintaining a closed-tube environment. The PCR requires <30 min (capillaries) or 1.5 h (96- or 384-well plates) and melting acquisition takes 1-2 min per capillary or 5 min per plate. PMID- 17401340 TI - In vivo models of proliferative vitreoretinopathy. AB - We outline current in vitro and in vivo models for experimental proliferative vitreoretinopathy (PVR) and provide a detailed protocol of our standardized in vivo PVR model. PVR is the leading cause of failed surgical procedures for the correction of rhegmatogenous retinal detachment. The pathogenesis of this multifactorial condition is still not completely understood. Experimental models for PVR help us understand the factors that play a role in the pathogenesis of the disease process in a controlled manner and allow for reproducible preclinical assessment of novel therapeutic interventions. We describe a cell injection model in detail that uses homologous retinal pigment epithelial (RPE) cell cultures to induce PVR over a 2-8 week period. PMID- 17401341 TI - Photochemical determination of different DNA structures. AB - The various conformations of DNA are thought to have important biological roles. Investigation of the local DNA conformational changes associated with biological events is therefore essential to an understanding of the functions of DNA. We have reported the photoreactivities of 5-halouracil in the five characteristic local DNA structures: the A, B and Z forms, protein-induced DNA kinks and the G quadruplex form. These studies demonstrate the detailed relationships between the local DNA structures and the photochemical products of photoinduced hydrogen abstraction by the resulting uracil-5-yl radicals, and show that this photochemical method can be used to detect DNA structures. Here, we describe in detail procedures that have been developed in our laboratory for probing DNA conformations by product analysis of photoirradiated 5-halouracil-containing DNA. The protocol includes the preparation of 5-halouracil-containing DNA and the characterization of the photoproducts, and it can be completed in 2 weeks. PMID- 17401342 TI - BAC 'landing' on chromosomes of Brachypodium distachyon for comparative genome alignment. AB - Fluorescence in situ hybridization (FISH) using bacterial artificial chromosomes (BACs) with large genomic DNA inserts as probes (BAC 'landing') is a powerful means by which eukaryotic genomes can be physically mapped and compared. Here we report a BAC landing protocol that has been developed specifically for the weedy grass species Brachypodium distachyon, which has been adopted recently by the scientific community as an alternative model for the temperate cereals and grasses. The protocol describes the preparation of somatic and meiotic chromosome substrates for FISH, the labeling of BACs, a chromosome mapping strategy, empirical conditions for optimal in situ hybridization and stringency washing, the detection of probes and the capturing and processing of images. The expected outcome of the protocol is the specific assignment of BACs containing single-copy inserts to one of the five linkage groups of the genome of this species. Once somatic or meiotic material is available, the entire protocol can be completed in about 3 d. The protocol has been customized empirically for B. distachyon and its near relatives, but it can be adapted with minor modifications to diverse plant species. PMID- 17401343 TI - Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. AB - We present a fast protocol that can be used to obtain highly purified cultures of proliferating adult human and rat Schwann cells accessible for non-viral transfection methods. The use of enriched genetically modified adult Schwann cells is of interest in the context of autologous cell transplantation within nerve transplants for peripheral nerve repair. Cell preparation from pre degenerated adult peripheral nerves is described, together with the use of melanocyte growth medium plus forskolin, fibroblast growth factor-2 (FGF-2), pituitary extract and heregulin as a selective, serum-free culture medium and a subsequent cell enrichment step (cold jet). Proliferating adult Schwann cells can be efficiently genetically modified using optimized, non-viral electroporation protocols. The protocol results in Schwann cell cultures that are more than 90 95% pure, and transfection efficiencies vary depending on the initial cell constitution from 20 to 40%. The procedure takes up to 21 d, depending on the length of the pre-degeneration period. PMID- 17401344 TI - A method to enable the investigation of murine bronchial immune cells, their cytokines and mediators. AB - Innovative therapies for severe lung diseases (such as allergic and chronic asthma, chronic obstructive pulmonary disease or any type of lung cancer) require a detailed understanding of the cellular and immune processes in the lung. This protocol details a method to obtain the immune cells of the bronchi as well as the cytokines and mediators produced by these cells for further investigation. The broncho-alveolar lavage fluid (BALF) is taken by injecting physiological solution through the tracheal tube into the murine airways and carefully regained by winding up the connected syringe. After centrifugation, the resulting BALF supernatant can be stored for detection of cytokines or other mediators by enzyme linked immunosorbent assay or other methods; the resuspended cell pellet can also be used for flow cytometric analyses, to check cell viability and the level of apoptosis, as well as other applications. In addition, CD4+ T cells isolated from wild-type and genetically modified mice alone or along with other immunologically important cells such as T regulatory cells, which can be used to reconstitute immunodeficient mice, may be retrieved from the airways with this method. This protocol can be completed within 35 min. PMID- 17401345 TI - L-proline-catalyzed enantioselective one-pot cross-Mannich reaction of aldehydes. AB - This protocol describes a procedure for the synthesis of syn-beta-amino alpha substituted aldehydes, versatile intermediates in synthetic organic chemistry, via asymmetric, direct, one-pot, three-component, cross-Mannich reaction of two different aldehydes. The reaction consists of two steps; one is the formation of imine by the reaction of aldehyde and p-anisidine in the presence of Pro, and the second step is the enantioselective addition reaction of enamine generated from the other aldehyde and Pro with the imine generated in the first step. As the aldehyde easily racemizes, gamma-amino alcohol was isolated and characterized after reduction. The yield and diastereo- and enantioselectivities are generally excellent. It will take approximately 26 h to complete the protocol: 0.5 h to set up the reaction, 20.5 h for the reaction and 5 h for the isolation and purification. PMID- 17401346 TI - Detection of non-covalent protein interactions by 'intensity fading' MALDI-TOF mass spectrometry: applications to proteases and protease inhibitors. AB - Among the main objectives of biomedical and proteomic research is to identify non covalent interactions involving proteins. Here we provide a detailed protocol to apply matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry for such a purpose using proteases and protease inhibitors in complex biological samples. Our methodology is based on monitoring the reduction in intensity of inhibitors' mass spectrometric signals when their protease target is added to the MALDI sample. The versatility of the protocol permits the target to be added in a soluble form (direct protocol) or immobilized form (indirect protocol). The 'intensity fading' phenomenon is greatly favored when the binding assay is carried out in the sub-micromolar range and the interacting partners occur in mixtures of non-binding compounds. This protocol can be completed in 10 h, taking 20 or 30 min per sample to perform the mass spectrometric data acquisition, depending on whether a soluble or an immobilized target is used. PMID- 17401347 TI - A primary culture technique of adult retina for regeneration studies on adult CNS neurons. AB - This protocol details a tissue culture technique that allows for quantified regeneration studies on adult retinal ganglion cells (RGCs), that is, CNS neurons. The method may also allow for elucidation of molecular cues, for example of signals relevant in neuronal survival and axon regeneration. The procedure relies on fractioned stripe culture of previously injured retina in defined culture media. Naive dendritic cell contacts of RGCs are preserved, and the system is independent of growth factors. In contrast to other techniques, the protocol is based on tissue grown from adult animals; it dispenses immature co cultures and evaluates the outgrowth of unmyelinated neurites in a milieu lacking CNS myelin. The technique is suitable for rodent retina from mouse or rat. A growth-conditioning injury of the optic nerve is set 10 days before retinal explantation. Explants are cultured for 5-7 days. Mere preparation of a single retina should be completed within 20 min. PMID- 17401348 TI - Protocol for the MPTP mouse model of Parkinson's disease. AB - This protocol describes our method of producing a reliable mouse model of Parkinson's disease (PD) using the neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). We discuss the particulars of the model, provide key references and outline what investigators need to know to develop the MPTP mouse model of PD safely and successfully. Completion of this protocol depends on the regimen of MPTP used and on the actual planned studies, which often range from 7 to 30 d. This protocol calls for implementation of safety measures and for the acquisition of several pieces of equipment, which are a one-time investment worth making if one elects to use this model on a regular basis. PMID- 17401349 TI - Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity. AB - Dissociated primary sensory neurons are commonly used to study growth factor dependent cell survival, axon outgrowth, differentiation and basic mechanisms of sensory physiology and pain. Spinal or trigeminal sensory neurons can be collected from embryos, neonates or adults, treated with enzymes that degrade the extracellular matrix, triturated and grown in defined media with or without growth factors and additional animal sera. Production of cultures can take as little as 2.5 h. Cells can be used almost immediately or maintained for as long as 1 month. Ease of production and the ability to control growth conditions make sensory neuron culture a powerful model system for studying basic neurobiology of central and peripheral nervous systems. PMID- 17401350 TI - Synthesis and application of an azobenzene amino acid as a light-switchable turn element in polypeptides. AB - The synthesis of an azobenzene amino acid (aa) for use as a photo-inducible conformational switch in polypeptides is described. The compound can be easily incorporated into an aa sequence by solid-phase peptide synthesis using standard 9-fluorenylmethoxycarbonyl methods. A reversible conformational change of the peptide backbone is induced by switching between the cis and trans configurations of the azobenzene moiety by irradiation with light of suitable wavelength. Thermal cis --> trans isomerization of this azobenzene aa is slow, enabling detailed structural investigations of the modified peptides, e.g., using NMR techniques. The total time for the synthesis of the photoswitch is typically 4 d, with an overall yield of 40-50%. PMID- 17401351 TI - The tagged microarray marker (TAM) method for high-throughput detection of single nucleotide and indel polymorphisms. AB - The tagged microarray marker (TAM) method allows high-throughput differentiation between predicted alternative PCR products. Typically, the method is used as a molecular marker approach to determining the allelic states of single nucleotide polymorphisms (SNPs) or insertion-deletion (indel) alleles at genomic loci in multiple individuals. Biotin-labeled PCR products are spotted, unpurified, onto a streptavidin-coated glass slide and the alternative products are differentiated by hybridization to fluorescent detector oligonucleotides that recognize corresponding allele-specific tags on the PCR primers. The main attractions of this method are its high throughput (thousands of PCRs are analyzed per slide), flexibility of scoring (any combination, from a single marker in thousands of samples to thousands of markers in a single sample, can be analyzed) and flexibility of scale (any experimental scale, from a small lab setting up to a large project). This protocol describes an experiment involving 3,072 PCRs scored on a slide. The whole process from the start of PCR setup to receiving the data spreadsheet takes 2 d. PMID- 17401352 TI - Establishment of 3D organotypic cultures using human neonatal epidermal cells. AB - This protocol describes an ex vivo three-dimensional coculture system optimized to study the skin regenerative ability of primary human keratinocytes grown at the air-liquid interface on collagen matrices embedded with human dermal fibroblasts. An option for enrichment of keratinocyte stem cells and their progeny using fluorescence-activated cell sorting is also provided. Initially, dermal equivalents, comprising human passaged fibroblasts seeded in a collagen matrix, are grown on porous filters (3 mum) placed in transwells. After 1 week, primary human keratinocytes are seeded on this base. One week later, an air-lift transition is performed, leading to the differentiation of the keratinocytes, which are macroscopically visible as artificial skin after a couple of days. The cultures can be harvested 1 week after the air-lift and processed for immunohistochemistry or gene expression analysis. The overall procedure can be completed in 3 weeks, including the preparation of the dermal equivalent and the seeding of the primary keratinocytes. PMID- 17401353 TI - Purification of naturally occurring peptides by reversed-phase HPLC. AB - Reversed-phase high performance liquid chromatography (HPLC) has become the method of choice for the purification of peptides and small proteins (M(r) < 10,000 Da) from natural sources. The technique combines high resolution and recovery with ease and speed of operation and is applicable to a wide range of peptides with different physicochemical properties. This protocol describes procedures for (1) the extraction of a biologically active peptide from animal tissue, (2) concentration of the extracts and partial purification on Sep-Pak cartridges, and (3) purification to near homogeneity on a range of silica-based HPLC columns. Standard operating procedures involve acetonitrile as organic modifier, trifluoroacetic acid as ion-pairing reagent and sequential chromatographies on octadecyl (C18), butyl (C4) and diphenyl wide-pore (300 A) columns under gradient elution conditions. The limiting factor in the time taken to isolate a peptide is usually the speed at which assays to detect the peptide can be performed, but purifications can generally be accomplished within 1 or 2 weeks. PMID- 17401354 TI - Purification of recombinant membrane proteins tagged with calmodulin-binding domains by affinity chromatography on calmodulin-agarose: example of nicotinamide nucleotide transhydrogenase. AB - This protocol describes affinity purification of bacterially expressed, recombinant membrane proteins fused with calmodulin-binding domains. As exemplified by the Escherichia coli nicotinamide nucleotide transhydrogenase, this method allows isolation of the protein fusions in a single chromatography step using elution with the calcium chelating agent EDTA and, unlike purification of His-tagged proteins on nickel chelate, it is not sensitive to the presence of strong reducing agents (e.g., DTT). Our protocol involves disruption of host bacteria by sonication, sedimentation of membranes by differential centrifugation, solubilization of membrane proteins and affinity chromatography on calmodulin-agarose. To achieve maximum purity and yield, the use of a combination of non-ionic and anionic detergents is suggested. Purification takes two working days, with an overnight wash of the column to increase the purity of the product. PMID- 17401355 TI - Synthesis of azobenzene-tethered DNA for reversible photo-regulation of DNA functions: hybridization and transcription. AB - A phosphoramidite monomer bearing an azobenzene is synthesized from D-threoninol. Using this monomer, azobenzene moieties can be introduced into oligodeoxyribonucleotide (DNA) at any position on a conventional DNA synthesizer. With this azobenzene-tethered DNA, formation and dissociation of a DNA duplex can be reversibly photo-regulated by cis-trans isomerization of the azobenzene. When the azobenzene takes a trans-form, a stable duplex is formed. After isomerization of the trans-azobenzene to its cis-form by UV-light irradiation (300 nm < lambda < 400 nm), the duplex can be dissociated into two strands. The duplex is reformed on photo-induced cis-trans isomerization (lambda > 400 nm). The introduction of azobenzenes into the T7 promoter at specific positions also efficiently and reversibly photo-regulates transcription by T7-RNA polymerase. The reversible regulation can be repeated many times without causing damage to the DNA or the azobenzene moiety. These procedures take approximately 10 d to complete. PMID- 17401356 TI - Metal-chelate affinity precipitation of proteins using responsive polymers. AB - Affinity precipitation of proteins uses polymers capable of reversible soluble insoluble transitions in response to small environmental changes (temperature, pH or solvent composition). Here we describe protocols for (i) the synthesis of responsive polymers with specific affinity to target proteins and (ii) the purification of proteins using these polymers. The purification is based on precipitation of the affinity complex between the protein and the polymer, which is induced by environmental changes. This separation strategy is simpler and more cost effective than conventional affinity column chromatography. Specifically, we describe the synthesis of thermoresponsive 1-vinylimidazole:N-isopropylacrylamide copolymers. The whole procedure takes 2-3 h when applied to purification of recombinant His-tag proteins or proteins with natural metal binding groups by means of metal chelate affinity precipitation. Optimization of the polymer composition and the type of chelating ions allows for target protein yields of 80% and higher. PMID- 17401357 TI - Quantification of F2-isoprostanes as a biomarker of oxidative stress. AB - Oxidant stress has been implicated in a wide variety of disease processes. One method to quantify oxidative injury is to measure lipid peroxidation. Quantification of a group of prostaglandin F(2alpha)-like compounds derived from the nonezymatic oxidation of arachidonic acid, termed the F2-isoprostanes (F2 IsoPs), provides an accurate assessment of oxidative stress both in vitro and in vivo. In fact, in a recent independent study sponsored by the National Institutes of Health (NIH), F2-IsoPs were shown to be the most reliable index of in vivo oxidant stress when compared against other well known biomarkers. This protocol details our laboratory's method to quantify F2-IsoPs in biological fluids and tissues using gas chromatography-mass spectrometry (GC-MS). This procedure can be completed for 12-15 samples in 6-8 h. PMID- 17401358 TI - Synthesis and use of an in-solution ratiometric fluorescent viscosity sensor. AB - A procedure for the synthesis of a ratiometric viscosity fluorescent sensor is described in this protocol. The essential requirement for the design of this sensor is the attachment of a primary fluorophore that has both a viscosity independent fluorescence emission (coumarin dye shown in blue) and an emission from a fluorophore that exhibits viscosity-dependent fluorescent quantum yield (p amino cinnamonitrile dye shown in red). The use of sensor 1 in viscosity measurements involves solubilization in a liquid of interest and excitation of the primary fluorophore at lambda(ex) = 360 nm. The secondary fluorophore is simultaneously excited via resonance energy transfer. The ratio of the fluorescent emission of the secondary over the primary fluorophore provides a fast and precise measurement of the viscosity of the solvent. The synthesis of compound 1 using commercially available materials can be completed within 5 d. PMID- 17401359 TI - Heteroduplex analysis by capillary array electrophoresis for rapid mutation detection in large multiexon genes. AB - Heteroduplex analysis (HA) has proven to be a robust tool for mutation detection. HA by capillary array electrophoresis (HA-CAE) was developed to increase throughput and allow the scanning of large multiexon genes in multicapillary DNA sequencers. HA-CAE is a straightforward and high-throughput technique to detect both known and novel DNA variants with a high level of sensitivity and specificity. It consists of only three steps: multiplex-PCR using fluorescently labeled primers, heteroduplex formation and electrophoresis in a multicapillary DNA sequencer. It allows, e.g., the complete coding and flanking intronic sequences of BRCA1 and BRCA2 genes from two patients (approximately 25 kb each) to be scanned in a single run of a 16-capillary sequencer, and has enabled us to detect 150 different mutations to date (both single nucleotide substitutions, or SNSs, and small insertions/deletions). Here, we describe the protocol developed in our laboratory to scan BRCA1, BRCA2, MLH1, MSH2 and MSH6 genes using an ABI3130XL sequencer. This protocol could be adapted to other instruments or to the study of other large multiexon genes and can be completed in 7-8 h. PMID- 17401360 TI - Bacterial glycosidases for the production of universal red blood cells. AB - Enzymatic removal of blood group ABO antigens to develop universal red blood cells (RBCs) was a pioneering vision originally proposed more than 25 years ago. Although the feasibility of this approach was demonstrated in clinical trials for group B RBCs, a major obstacle in translating this technology to clinical practice has been the lack of efficient glycosidase enzymes. Here we report two bacterial glycosidase gene families that provide enzymes capable of efficient removal of A and B antigens at neutral pH with low consumption of recombinant enzymes. The crystal structure of a member of the alpha-N-acetylgalactosaminidase family reveals an unusual catalytic mechanism involving NAD+. The enzymatic conversion processes we describe hold promise for achieving the goal of producing universal RBCs, which would improve the blood supply while enhancing the safety of clinical transfusions. PMID- 17401361 TI - High-throughput identification of transcription start sites, conserved promoter motifs and predicted regulons. AB - Using 62 probe-level datasets obtained with a custom-designed Caulobacter crescentus microarray chip, we identify transcriptional start sites of 769 genes, 53 of which are transcribed from multiple start sites. Transcriptional start sites are identified by analyzing probe signal cross-correlation matrices created from probe pairs tiled every 5 bp upstream of the genes. Signals from probes binding the same message are correlated. The contribution of each promoter for genes transcribed from multiple promoters is identified. Knowing the transcription start site enables targeted searching for regulatory-protein binding motifs in the promoter regions of genes with similar expression patterns. We identified 27 motifs, 17 of which share no similarity to the characterized motifs of other C. crescentus transcriptional regulators. Using these motifs, we predict coregulated genes. We verified novel promoter motifs that regulate stress response genes, including those responding to uranium challenge, a stress response sigma factor and a stress-response noncoding RNA. PMID- 17401362 TI - C. elegans mitochondrial factor WAH-1 promotes phosphatidylserine externalization in apoptotic cells through phospholipid scramblase SCRM-1. AB - Externalization of phosphatidylserine, which is normally restricted to the inner leaflet of plasma membrane, is a hallmark of mammalian apoptosis. It is not known what activates and mediates the phosphatidylserine externalization process in apoptotic cells. Here, we report the development of an annexin V-based phosphatidylserine labelling method and show that a majority of apoptotic germ cells in Caenorhabditis elegans have surface-exposed phosphatidylserine, indicating that phosphatidylserine externalization is a conserved apoptotic event in worms. Importantly, inactivation of the gene encoding either the C. elegans apoptosis-inducing factor (AIF) homologue (WAH-1), a mitochondrial apoptogenic factor, or the C. elegans phospholipid scramblase 1 (SCRM-1), a plasma membrane protein, reduces phosphatidylserine exposure on the surface of apoptotic germ cells and compromises cell-corpse engulfment. WAH-1 associates with SCRM-1 and activates its phospholipid scrambling activity in vitro. Thus WAH-1, after its release from mitochondria during apoptosis, promotes plasma membrane phosphatidylserine externalization through its downstream effector, SCRM-1. PMID- 17401363 TI - Genome-wide association study of prostate cancer identifies a second risk locus at 8q24. AB - Recently, common variants on human chromosome 8q24 were found to be associated with prostate cancer risk. While conducting a genome-wide association study in the Cancer Genetic Markers of Susceptibility project with 550,000 SNPs in a nested case-control study (1,172 cases and 1,157 controls of European origin), we identified a new association at 8q24 with an independent effect on prostate cancer susceptibility. The most significant signal is 70 kb centromeric to the previously reported SNP, rs1447295, but shows little evidence of linkage disequilibrium with it. A combined analysis with four additional studies (total: 4,296 cases and 4,299 controls) confirms association with prostate cancer for rs6983267 in the centromeric locus (P = 9.42 x 10(-13); heterozygote odds ratio (OR): 1.26, 95% confidence interval (c.i.): 1.13-1.41; homozygote OR: 1.58, 95% c.i.: 1.40-1.78). Each SNP remained significant in a joint analysis after adjusting for the other (rs1447295 P = 1.41 x 10(-11); rs6983267 P = 6.62 x 10( 10)). These observations, combined with compelling evidence for a recombination hotspot between the two markers, indicate the presence of at least two independent loci within 8q24 that contribute to prostate cancer in men of European ancestry. We estimate that the population attributable risk of the new locus, marked by rs6983267, is higher than the locus marked by rs1447295 (21% versus 9%). PMID- 17401364 TI - Multiple regions within 8q24 independently affect risk for prostate cancer. AB - After the recent discovery that common genetic variation in 8q24 influences inherited risk of prostate cancer, we genotyped 2,973 SNPs in up to 7,518 men with and without prostate cancer from five populations. We identified seven risk variants, five of them previously undescribed, spanning 430 kb and each independently predicting risk for prostate cancer (P = 7.9 x 10(-19) for the strongest association, and P < 1.5 x 10(-4) for five of the variants, after controlling for each of the others). The variants define common genotypes that span a more than fivefold range of susceptibility to cancer in some populations. None of the prostate cancer risk variants aligns to a known gene or alters the coding sequence of an encoded protein. PMID- 17401365 TI - Impaired microRNA processing enhances cellular transformation and tumorigenesis. AB - MicroRNAs (miRNAs) are a new class of small noncoding RNAs that post transcriptionally regulate the expression of target mRNA transcripts. Many of these target mRNA transcripts are involved in proliferation, differentiation and apoptosis, processes commonly altered during tumorigenesis. Recent work has shown a global decrease of mature miRNA expression in human cancers. However, it is unclear whether this global repression of miRNAs reflects the undifferentiated state of tumors or causally contributes to the transformed phenotype. Here we show that global repression of miRNA maturation promotes cellular transformation and tumorigenesis. Cancer cells expressing short hairpin RNAs (shRNAs) targeting three different components of the miRNA processing machinery showed a substantial decrease in steady-state miRNA levels and a more pronounced transformed phenotype. In animals, miRNA processing-impaired cells formed tumors with accelerated kinetics. These tumors were more invasive than control tumors, suggesting that global miRNA loss enhances tumorigenesis. Furthermore, conditional deletion of Dicer1 enhanced tumor development in a K-Ras-induced mouse model of lung cancer. Overall, these studies indicate that abrogation of global miRNA processing promotes tumorigenesis. PMID- 17401366 TI - Genome-wide association study identifies a second prostate cancer susceptibility variant at 8q24. AB - Prostate cancer is the most prevalent noncutaneous cancer in males in developed regions, with African American men having among the highest worldwide incidence and mortality rates. Here we report a second genetic variant in the 8q24 region that, in conjunction with another variant we recently discovered, accounts for about 11%-13% of prostate cancer cases in individuals of European descent and 31% of cases in African Americans. We made the current discovery through a genome wide association scan of 1,453 affected Icelandic individuals and 3,064 controls using the Illumina HumanHap300 BeadChip followed by four replication studies. A key step in the discovery was the construction of a 14-SNP haplotype that efficiently tags a relatively uncommon (2%-4%) susceptibility variant in individuals of European descent that happens to be very common (approximately 42%) in African Americans. The newly identified variant shows a stronger association with affected individuals who have an earlier age at diagnosis. PMID- 17401367 TI - Interaction of the selectin ligand PSGL-1 with chemokines CCL21 and CCL19 facilitates efficient homing of T cells to secondary lymphoid organs. AB - P-selectin glycoprotein ligand 1 (PSGL-1) is central to the trafficking of immune effector cells to areas of inflammation through direct interactions with P selectin, E-selectin and L-selectin. Here we show that PSGL-1 was also required for efficient homing of resting T cells to secondary lymphoid organs but functioned independently of selectin binding. PSGL-1 mediated an enhanced chemotactic T cell response to the secondary lymphoid organ chemokines CCL21 and CCL19 but not to CXCL12 or to inflammatory chemokines. Our data show involvement of PSGL-1 in facilitating the entry of T cells into secondary lymphoid organs, thereby demonstrating the bifunctional nature of this molecule. PMID- 17401368 TI - Local increase in thymic stromal lymphopoietin induces systemic alterations in B cell development. AB - The cytokine thymic stromal lymphopoietin (TSLP) drives immature B cell development in vitro and may regulate T helper type 2 responses. Here we analyzed the involvement of TSLP in B cell development in vivo with a doxycycline inducible, keratin 5-driven transgene encoding TSLP (K5-TSLP). K5-TSLP-transgenic mice given doxycycline showed an influx of immature B cells into the periphery, with population expansion of follicular mature B cells, near-complete loss of marginal zone and marginal zone precursor B cells, and 'preferential' population expansion of peritoneal B-1b B cells. These changes promoted cryoglobulin production and immune complex-mediated renal disease. Identical events occurred in mice without T cells, in alternative TSLP-transgenic models and in K5-TSLP transgenic mice with undetectable systemic TSLP. These observations suggest that signals mediating localized TSLP expression may modulate systemic B cell development and promote humoral autoimmunity. PMID- 17401369 TI - Image-based multivariate profiling of drug responses from single cells. AB - Quantitative analytical approaches for discovering new compound mechanisms are required for summarizing high-throughput, image-based drug screening data. Here we present a multivariate method for classifying untreated and treated human cancer cells based on approximately 300 single-cell phenotypic measurements. This classification provides a score, measuring the magnitude of the drug effect, and a vector, indicating the simultaneous phenotypic changes induced by the drug. These two quantities were used to characterize compound activities and identify dose-dependent multiphasic responses. A systematic survey of profiles extracted from a 100-compound compendium of image data revealed that only 10-15% of the original features were required to detect a compound effect. We report the most informative image features for each compound and fluorescence marker set using a method that will be useful for determining minimal collections of readouts for drug screens. Our approach provides human-interpretable profiles and automatic determination of on- and off-target effects. PMID- 17401370 TI - Imaging dynamics of endogenous mitochondrial RNA in single living cells. AB - We developed genetically encoded RNA probes for characterizing localization and dynamics of mitochondrial RNA (mtRNA) in single living cells. The probes consist of two RNA-binding domains of PUMILIO1, each connected with split fragments of a fluorescent protein capable of reconstituting upon binding to a target RNA. We designed the probes to specifically recognize a 16-base sequence of mtRNA encoding NADH dehydrogenase subunit 6 (ND6) and to be targeted into the mitochondrial matrix, which allowed real-time imaging of ND6 mtRNA localization in living cells. We showed that ND6 mtRNA is localized within mitochondria and concentrated particularly on mitochondrial DNA (mtDNA). Movement of the ND6 mtRNA is restricted but oxidative stress induces the mtRNA to disperse in the mitochondria and gradually decompose. These probes provide a means to study spatial and temporal mRNA dynamics in intracellular compartments in living mammalian cells. PMID- 17401371 TI - RNA visualization in live bacterial cells using fluorescent protein complementation. AB - We describe a technique for the detection and localization of RNA transcripts in living cells. The method is based on fluorescent-protein complementation regulated by the interaction of a split RNA-binding protein with its corresponding RNA aptamer. In our design, the RNA-binding protein is the eukaryotic initiation factor 4A (eIF4A). eIF4A is dissected into two fragments, and each fragment is fused to split fragments of the enhanced green fluorescent protein (EGFP). Coexpression of the two protein fusions in the presence of a transcript containing eIF4A-interacting RNA aptamer resulted in the restoration of EGFP fluorescence in Escherichia coli cells. We also applied this technique to the visualization of an aptamer-tagged mRNA and 5S ribosomal RNA (rRNA). We observed distinct spatial and temporal changes in fluorescence within single cells, reflecting the nature of the transcript. PMID- 17401372 TI - Structural basis for autoinhibition of Notch. AB - Notch receptors transmit signals between adjacent cells. Signaling is initiated when ligand binding induces metalloprotease cleavage of Notch within an extracellular negative regulatory region (NRR). We present here the X-ray structure of the human NOTCH2 NRR, which adopts an autoinhibited conformation. Extensive interdomain interactions within the NRR bury the metalloprotease site, showing that a substantial conformational movement is necessary to expose this site during activation by ligand. Leukemia-associated mutations in NOTCH1 probably release autoinhibition by destabilizing the conserved hydrophobic core of the NRR. PMID- 17401373 TI - Potent effect of target structure on microRNA function. AB - MicroRNAs (miRNAs) are small noncoding RNAs that repress protein synthesis by binding to target messenger RNAs. We investigated the effect of target secondary structure on the efficacy of repression by miRNAs. Using structures predicted by the Sfold program, we model the interaction between an miRNA and a target as a two-step hybridization reaction: nucleation at an accessible target site followed by hybrid elongation to disrupt local target secondary structure and form the complete miRNA-target duplex. This model accurately accounts for the sensitivity to repression by let-7 of various mutant forms of the Caenorhabditis elegans lin 41 3' untranslated region and for other experimentally tested miRNA-target interactions in C. elegans and Drosophila melanogaster. These findings indicate a potent effect of target structure on target recognition by miRNAs and establish a structure-based framework for genome-wide identification of animal miRNA targets. PMID- 17401374 TI - The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. AB - MicroRNAs (miRNAs) are endogenous noncoding RNAs, about 22 nucleotides in length, that mediate post-transcriptional gene silencing by annealing to inexactly complementary sequences in the 3'-untranslated regions of target mRNAs. Our current understanding of the functions of miRNAs relies mainly on their tissue specific or developmental stage-dependent expression and their evolutionary conservation, and therefore is primarily limited to their involvement in developmental regulation and oncogenesis. Of more than 300 miRNAs that have been identified, miR-1 and miR-133 are considered to be muscle specific. Here we show that miR-1 is overexpressed in individuals with coronary artery disease, and that when overexpressed in normal or infarcted rat hearts, it exacerbates arrhythmogenesis. Elimination of miR-1 by an antisense inhibitor in infarcted rat hearts relieved arrhythmogenesis. miR-1 overexpression slowed conduction and depolarized the cytoplasmic membrane by post-transcriptionally repressing KCNJ2 (which encodes the K(+) channel subunit Kir2.1) and GJA1 (which encodes connexin 43), and this likely accounts at least in part for its arrhythmogenic potential. Thus, miR-1 may have important pathophysiological functions in the heart, and is a potential antiarrhythmic target. PMID- 17401375 TI - Helicobacter pylori infection triggers aberrant expression of activation-induced cytidine deaminase in gastric epithelium. AB - Infection with Helicobacter pylori (H. pylori) is a risk factor for the development of gastric cancer. Here we show that infection of gastric epithelial cells with 'cag' pathogenicity island (cagPAI)-positive H. pylori induced aberrant expression of activation-induced cytidine deaminase (AID), a member of the cytidine-deaminase family that acts as a DNA- and RNA-editing enzyme, via the IkappaB kinase-dependent nuclear factor-kappaB activation pathway. H. pylori mediated upregulation of AID resulted in the accumulation of nucleotide alterations in the TP53 tumor suppressor gene in gastric cells in vitro. Our findings provide evidence that aberrant AID expression caused by H. pylori infection might be a mechanism of mutation accumulation in the gastric mucosa during H. pylori-associated gastric carcinogenesis. PMID- 17401376 TI - Direct suppression of CNS autoimmune inflammation via the cannabinoid receptor CB1 on neurons and CB2 on autoreactive T cells. AB - The cannabinoid system is immunomodulatory and has been targeted as a treatment for the central nervous system (CNS) autoimmune disease multiple sclerosis. Using an animal model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE), we investigated the role of the CB(1) and CB(2) cannabinoid receptors in regulating CNS autoimmunity. We found that CB(1) receptor expression by neurons, but not T cells, was required for cannabinoid-mediated EAE suppression. In contrast, CB(2) receptor expression by encephalitogenic T cells was critical for controlling inflammation associated with EAE. CB(2)-deficient T cells in the CNS during EAE exhibited reduced levels of apoptosis, a higher rate of proliferation and increased production of inflammatory cytokines, resulting in severe clinical disease. Together, our results demonstrate that the cannabinoid system within the CNS plays a critical role in regulating autoimmune inflammation, with the CNS directly suppressing T-cell effector function via the CB(2) receptor. PMID- 17401377 TI - Angiogenic inhibition reduces germinal matrix hemorrhage. AB - The germinal matrix of premature infants is selectively vulnerable to hemorrhage within the first 48 h of life. To assess the role of vascular immaturity in germinal matrix hemorrhage (GMH), we evaluated germinal matrix angiogenesis in human fetuses and premature infants, as well as in premature rabbit pups, and noted active vessel remodeling in all three. Vascular endothelial growth factor (VEGF), angiopoietin-2 and endothelial cell proliferation were present at consistently higher levels in the germinal matrix relative to the white matter anlagen and cortical mantle. On that basis, we asked whether prenatal treatment with either of two angiogenic inhibitors, the COX-2 inhibitor celecoxib, or the VEGFR2 inhibitor ZD6474, could suppress the incidence of GMH in premature rabbit pups. Celecoxib treatment decreased angiopoietin-2 and VEGF levels as well as germinal matrix endothelial proliferation. Furthermore, treatment with celecoxib or ZD6474 substantially decreased the incidence of GMH. Thus, by suppressing germinal matrix angiogenesis, prenatal celecoxib or ZD6474 treatment may be able to reduce both the incidence and severity of GMH in susceptible premature infants. PMID- 17401378 TI - The Cfd1-Nbp35 complex acts as a scaffold for iron-sulfur protein assembly in the yeast cytosol. AB - Biogenesis of iron-sulfur ([Fe-S]) proteins in eukaryotes requires the function of complex proteinaceous machineries in both mitochondria and cytosol. In contrast to the mitochondrial pathway, little is known about [Fe-S] protein assembly in the cytosol. So far, four highly conserved proteins (Cfd1, Nbp35, Nar1 and Cia1) have been identified as members of the cytosolic [Fe-S] protein assembly machinery, but their molecular function is unresolved. Using in vivo and in vitro approaches, we found that the soluble P-loop NTPases Cfd1 and Nbp35 form a complex and bind up to three [4Fe-4S] clusters, one at the N terminus of Nbp35 and one each at a new C-terminal cysteine-rich motif present in both proteins. These labile [Fe-S] clusters can be rapidly transferred and incorporated into target [Fe-S] apoproteins in a Nar1- and Cia1-dependent fashion. Our data suggest that the Cfd1-Nbp35 complex functions as a novel scaffold for [Fe-S] cluster assembly in the eukaryotic cytosol. PMID- 17401379 TI - Molecular imaging of hydrogen peroxide produced for cell signaling. AB - Hydrogen peroxide (H2O2) is emerging as a newly recognized messenger in cellular signal transduction. However, a substantial challenge in elucidating its diverse roles in complex biological environments is the lack of methods for probing this reactive oxygen metabolite in living systems with molecular specificity. Here we report the synthesis and application of Peroxy Green 1 (PG1) and Peroxy Crimson 1 (PC1), two new fluorescent probes that show high selectivity for H2O2 and are capable of visualizing endogenous H2O2 produced in living cells by growth factor stimulation, including the first direct imaging of peroxide produced for brain cell signaling. The combined features of reactive oxygen species selectivity, sensitivity to signaling levels of H2O2, and live-cell compatibility presage many new opportunities for PG1, PC1 and related synthetic reagents for exploring the physiological roles of H2O2 in living systems with molecular imaging. PMID- 17401383 TI - New ways to arrest phagosome maturation. PMID- 17401384 TI - Dangerous liaisons: polyglutamine meets HMGB. PMID- 17401385 TI - NF-kappaB links oestrogen receptor signalling and EMT. PMID- 17401386 TI - The double (strand break) life of Par-3. PMID- 17401387 TI - Rapid cargo shuttling by caveolae. PMID- 17401390 TI - Retractions. PMID- 17401392 TI - Human herpesvirus-6 encephalitis following allogeneic hematopoietic stem cell transplantation. AB - Immunosuppressive monoclonal antibodies directed to immune system cells may reduce rejection and graft versus host disease (GvHD) after allogeneic stem cell transplantation (SCT), but can increase the risks of viral infection. Here, we report human herpes virus-6 (HHV-6) encephalitis despite antiviral prophylaxis in 5 of 43 (11.6%) patients receiving alemtuzumab supported conditioning. Encephalitis occurred at 41-103 days (median 60 days) presenting with confusion in all patients, combined with amnesia (n=3) or seizures (n=2). MRI revealed non specific white matter changes in two and a non-enhancing medial temporal lobe lesion in three patients. Cerebrospinal fluid (CSF) PCR amplification for HHV-6 was positive in all five patients, (600-2 25 000 (median 4700) copies/ml CSF), while analysis of peripheral blood revealed 100-22 500 (median 1200) viral copies/ml plasma. CSF protein was elevated in four patients, with minimal CSF pleocytosis. Intravenous foscarnet produced neurological improvement at 8-13 (median 11) days and negative plasma PCR at 30-66 (median 50) days. Four patients had complete neurological recovery, but one patient with persistent viral DNA in the CSF succumbed to progressive encephalopathy. Given this high incidence of HHV 6 and the possibility of successful outcome with prompt treatment, a high index of suspicion of this disorder is required in recipients of monoclonal antibody supported allografts. PMID- 17401393 TI - Long-term renal function following bone marrow transplantation. AB - Renal function, evaluated as glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), was investigated in 187 pediatric patients who underwent allogeneic (n=169) or autologous bone marrow transplantation (BMT). Allogeneic BMT patients were divided into three groups: hematological malignancies, aplastic anemia and non-malignant diseases, whereas autologous patients constituted a fourth group. A total of 64% received total body irradiation (TBI) as conditioning therapy, and 50 healthy children served as controls. GFR and ERPF were normal before transplantation. After 1 year, both GFR and ERPF were significantly reduced. GFR had recovered slightly after 3 years and remained stable thereafter. Recovery in ERPF was not apparent. Renal impairment was found in 41% of patients at 1 year, in 31% at 3 years and in 11% 7 years after BMT. Patients with hematological malignancies had lower GFRs than patients with non malignant diseases at all time points. The most important risk factor as regards chronic renal impairment was TBI. Type of donor, cyclophosphamide (CY), or acute graft-versus-host disease (GVHD) did not seem to contribute to the development of chronic renal impairment. We suggest that tests of renal function should be included in long-term followup after BMT. PMID- 17401394 TI - Early relapses do not impact survival after autologous stem cell transplantation in African Americans with multiple myeloma. PMID- 17401395 TI - Invasive Pseudomonas aeruginosa infections: high rate of recurrence and mortality after hematopoietic cell transplantation. AB - Limited data exist regarding the incidence and factors associated with outcome of invasive Pseudomonal infections in hematopoietic cell transplant (HCT). A retrospective analysis of cases of invasive Pseudomonas aeruginosa infection and factors associated with outcome was performed. P. aeruginosa invasive infection occurred in 95 of 5772 patients (1.65%) a median of 63 days after HCT (range 5 1435). Only 28% of infections occurred during periods of neutropenia (absolute neutrophil count<500 cells/mm(3)). Infection-attributable mortality during the initial episode of infection was 35.8%. Factors associated with initial mortality included the presence of a copathogen and high-dose steroid use. Ten (16.4%) of those who survived the initial infection experienced a recurrence of P. aeruginosa infection at a median of 9 days (range 3-17) after stopping antibiotics and 60% of those died as a result of recurrent infection a median of 1 day (range 1-7) after onset of recurrence. Grade 3-4 graft-versus-host disease was associated with a higher risk of recurrent infection. The risk of recurrence was not influenced by the presence of copathogens. Thus, invasive P. aeruginosa infections are associated with high recurrence rates and mortality in this immunocompromised population. Aggressive attempts to reduce immunosuppression and to treat copathogens may help during the initial infection. PMID- 17401396 TI - Impact of cumulative anthracycline dose, preparative regimen and chronic graft versus-host disease on pulmonary and cardiac function in children 5 years after allogeneic hematopoietic stem cell transplantation: a prospective evaluation on behalf of the EBMT Pediatric Diseases and Late Effects Working Parties. AB - This prospective study focused on risk factors and clinical outcome of pulmonary and cardiac late effects after allogeneic hematopoietic stem cell transplantation (allo-HSCT). We prospectively evaluated 162 children by pulmonary function tests (PFTs) and cardiac shortening fraction (SF) before allo-HSCT and yearly up to the 5th year of follow-up. The 5-year cumulative incidence of lung and cardiac impairment was 35 (hazard rate=0.03) and 26% (hazard rate=0.06), respectively. Patients presenting abnormal PFTs and SF at last follow-up were 19 and 13%, respectively, with a median Lansky performance status of 90% (70-100). Chronic graft-versus-host disease (c-GVHD) was the major risk factor for reduced lung function in univariate (P=0.02) and multivariate analysis (P=0.02). Total body irradiation (TBI) alone and TBI together with pre-transplant anthracycline administration were significant risk factors for reduced cardiac function in univariate analysis, only (P=0.04 and 0.004, respectively). In conclusion, our prospective study demonstrates an asymptomatic post-allo-HSCT deterioration of pulmonary and cardiac function in some long-term survivors, who had been transplanted in childhood, and thus emphasizes the need for lifelong cardiopulmonary monitoring and the development of new strategies both to reduce pre-transplant cardiotoxic regimens and to treat more efficiently c-GVHD. PMID- 17401397 TI - Targeting the stem cell niche: squeezing blood from bones. AB - During human development, stem cells establish themselves in specific anatomic locations or niches. The niche harbors the stem cells, and regulates how stem cells proliferate. The interaction between stem cells and their niche affects stem cell function, and offers an opportunity to improve the marrow microenvironment. Osteoblasts produce hematopoietic growth factors and are activated by parathyroid hormone (PTH). A calcium sensing receptor, expressed by hematopoietic stem cells, regulates the niche and can be targeted to increase stem cell numbers. Therefore, drugs that affect osteoblast function or target calcium receptors may be useful for stem cell mobilization and engraftment. In this review, the biology of the stem cell niche and the potential therapeutic manipulations of the stem cell niche are reviewed. PTH is in clinical trials for patients who have not mobilized autologous stem cells well. The limiting cell numbers for adult cord blood transplantation increase the risk of infection, and PTH is currently in a clinical trial following cord blood transplantation in an effort to improve engraftment and immune reconstitution. PMID- 17401398 TI - A case of JAK2 positive essential thrombocythemia 16.5 years after autologous marrow transplantation for AML. PMID- 17401399 TI - Motivation. PMID- 17401400 TI - On science and English. PMID- 17401401 TI - Precaution and analysis: two sides of the same coin? Introduction to talking point on the precautionary principle. PMID- 17401402 TI - The precautionary principle should not be used as a basis for decision-making. Talking point on the precautionary principle. PMID- 17401403 TI - Risk, precaution and science: towards a more constructive policy debate. Talking point on the precautionary principle. PMID- 17401404 TI - The human impact on biological diversity. How species adapt to urban challenges sheds light on evolution and provides clues about conservation. PMID- 17401405 TI - A breath of hope. Researchers are combining their efforts to study the genetic and environmental factors in asthma. PMID- 17401406 TI - The elusive determinants of replication origins. PMID- 17401407 TI - The emergence of shape: notions from the study of the Drosophila tracheal system. AB - The generation of bodies and body parts with specific shapes and sizes has been a longstanding issue in biology. Morphogenesis in general and organogenesis in particular are complex events that involve global changes in cell populations in terms of their proliferation, migration, differentiation and shape. Recent studies have begun to address how these synchronized changes are controlled by the genes that specify cell fate and by the ability of cells to respond to extracellular cues. In particular, a notable shift in this research has occurred owing to the ability to address these issues in the context of the whole organism. For such studies, the Drosophila tracheal system has proven to be a particularly appropriate model. Here, my aim is to highlight some ideas that have arisen through our studies, and those from other groups, of Drosophila tracheal development. Rather than providing an objective review of the features of tracheal development, I intend to discuss some selected notions that I think are relevant to the question of shape generation. PMID- 17401408 TI - Chaperoning ribonucleoprotein biogenesis in health and disease. AB - The survival motor neuron (SMN) protein is part of a macromolecular complex that functions in the biogenesis of small nuclear ribonucleoproteins (snRNPs)--the essential components of the pre-messenger RNA splicing machinery--as well as probably other RNPs. Reduced levels of SMN expression cause the inherited motor neuron disease spinal muscular atrophy (SMA). Knowledge of the composition, interactions and functions of the SMN complex has advanced greatly in recent years. The emerging picture is that the SMN complex acts as a macromolecular chaperone of RNPs to increase the efficiency and fidelity of RNA-protein interactions in vivo, and to provide an opportunity for these interactions to be regulated. In addition, it seems that RNA metabolism deficiencies underlie SMA. Here, a dual dysfunction hypothesis is presented in which two mechanistically and temporally distinct defects--that are dependent on the extent of SMN reduction in SMA--affect the homeostasis of specific messenger RNAs encoding proteins essential for motor neuron development and function. PMID- 17401411 TI - Regulating nanomedicine. PMID- 17401409 TI - Replication fork barriers: pausing for a break or stalling for time? AB - Defects in chromosome replication can lead to translocations that are thought to result from recombination events at stalled DNA replication forks. The progression of forks is controlled by an essential DNA helicase, which unwinds the parental duplex and can stall on encountering tight protein-DNA complexes. Such pause sites are hotspots for recombination and it has been proposed that stalled replisomes disassemble, leading to fork collapse. However, in both prokaryotes and eukaryotes it now seems that paused forks are surprisingly stable, so that DNA synthesis can resume without recombination if the barrier protein is removed. Recombination at stalled forks might require other events that occur after pausing, or might be dependent on features of the surrounding DNA sequence. These findings have important implications for our understanding of the regulation of genome stability in eukaryotic cells, in which pausing of forks is mediated by specific proteins that are associated with the replicative helicase. PMID- 17401414 TI - Spintronics. Change of direction. PMID- 17401412 TI - Nanomedicine: industry-wise research. PMID- 17401415 TI - Nanocomposites. Model interfaces. PMID- 17401416 TI - Biosensors. Plasmons spring into action. PMID- 17401417 TI - Polymer-nanoparticle films. Entropy and enthalpy at play. PMID- 17401418 TI - Material witness. Rollobots. PMID- 17401419 TI - Living with cracks: damage and repair in human bone. AB - Our bones are full of cracks, which form and grow as a result of daily loading activities. Bone is the major structural material in our bodies. Although weaker than many engineering materials, it has one trick that keeps it ahead - it can repair itself. Small cracks, which grow under cyclic stresses by the mechanism of fatigue, can be detected and removed before they become long enough to be dangerous. This article reviews the work that has been done to understand how cracks form and grow in bone, and how they can be detected and repaired in a timely manner. This is truly an interdisciplinary research field, requiring the close cooperation of materials scientists, biologists and engineers. PMID- 17401422 TI - Mutant p53: from guardian to fallen angel? PMID- 17401423 TI - p53 alterations in human cancer: more questions than answers. AB - The strongest and undisputed fact about p53 is the high frequency of p53 alterations in human cancer and that mutant p53 proteins constitute a complex family of several hundred proteins with heterogeneous properties. Beyond these observations, the p53 pathway and its regulation in a normal cell is like a desert trail, always moving with the wind of novel findings. The field is full of black boxes that are often ignored for sake of simplicity or because they do not fit with the current dominant view. Mutant p53 protein accumulation in tumours is the best example of a preconceived idea, as there is no experimental evidence to explain this observation. In this review, we will discuss several questions concerning the activity or selection of p53 mutations. The central domain of the p53 protein targeted by 80% of p53 mutations is associated with the DNA-binding activity of the p53 protein, but it is also the binding site for several proteins that play a key role in p53 regulation such as ASPP proteins or BclxL. The role of impaired DNA binding and/or protein interactions in tumour development has not been fully elucidated. Similarly, novel animal models carrying either missense p53 mutations or inducible p53 have provided abundant observations, some of which could challenge our view on p53 function as a tumour suppressor gene. Finally, the possible clinical applications of p53 will be discussed. PMID- 17401424 TI - TP53 mutations in human cancers: functional selection and impact on cancer prognosis and outcomes. AB - A large amount of data is available on the functional impact of missense mutations in TP53 and on mutation patterns in many different cancers. New data on mutant p53 protein function, cancer phenotype and prognosis have recently been integrated in the International Agency for Research on Cancer TP53 database (http://www-p53.iarc.fr/). Based on these data, we summarize here current knowledge on the respective roles of mutagenesis and biological selection of mutations with specific functional characteristic in shaping the patterns and phenotypes of mutations observed in human cancers. The main conclusion is that intrinsic mutagenicity rates, loss of transactivation activities, and to a lesser extent, dominant-negative activities are the main driving forces that determine TP53 mutation patterns and influence tumor phenotype. In contrast, current experimental data on the acquisition of oncogenic activities (gain of function) by p53 mutants are too scarce and heterogenous to assess whether this property has an impact on tumor development and outcome. In the case of inherited TP53 mutations causing Li-Fraumeni and related syndromes, the age at onset of some tumor types is in direct relation with the degree of loss of transactivation capacity of missense mutations. Finally, studies on large case series demonstrate that TP53 mutations are independent markers of bad prognosis in breast and several other cancers, and that the exact type and position of the mutation influences disease outcome. Further studies are needed to determine how TP53 haplotypes or loss of alleles interact with mutations to modulate their impact on cancer development and prognosis. PMID- 17401425 TI - TP53 mutations and hepatocellular carcinoma: insights into the etiology and pathogenesis of liver cancer. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and the major risk factors include chronic infections with the hepatitis B (HBV) or C (HCV) virus, and exposure to dietary aflatoxin B(1) (AFB(1)) or alcohol consumption. Multiple genetic and epigenetic changes are involved in the molecular pathogenesis of HCC, for example, somatic mutations in the p53 tumor suppressor gene (TP53) and the activation of the WNT signal transduction pathway. AFB(1) frequently induces G:C to T:A transversions at the third base in codon 249 of TP53 and cooperates with HBV in causing p53 mutations in HCC. The detection of TP53 mutant DNA in plasma is a biomarker of both AFB(1) exposure and HCC risk. Chronic infection with HBV and HCV viruses, and oxyradical disorders including hemochromatosis, also generate reactive oxygen/nitrogen species that can both damage DNA and mutate cancer-related genes such as TP53. Certain mutant p53 proteins may exhibit a 'gain of oncogenic function'. The p53 biological network is a key responder to this oxidative and nitrosative stress. Depending on the extent of the DNA damage, p53 regulates the transcription of protective antioxidant genes and with extensive DNA damage, transactivates pro-oxidant genes that contribute to apoptosis. The X gene of HBV (HBx) is the most common open reading frame integrated into the host genome in HCC and the integrated HBx is frequently mutated. Mutant HBx proteins still retain their ability to bind to p53, and attenuate DNA repair and p53-mediated apoptosis. In summary, both viruses and chemicals are implicated in the etiology of TP53 mutations during the molecular pathogenesis of HCC. PMID- 17401426 TI - Crippling p53 activities via knock-in mutations in mouse models. AB - The tumor suppressor p53 is the most frequently mutated gene in human cancer. In vivo models have been generated using knock-in alleles in which missense mutations are introduced that mimic the kinds of mutations found in human cancers, or that abolish specific p53 functions. Critically, these studies examine the in vivo and physiological functions of p53. Studies indicate that p53 missense mutations in the DNA-binding domain identical with those inherited in the Li-Fraumeni syndrome, have distinct properties. Studies in mice with mutants that separate cell-cycle arrest and apoptosis functions of p53 show delayed onset of tumor development, suggesting that both p53 functions are crucial for suppressing tumors. Mice with mutations at post-translational modification sites exhibit subtle effects on p53 activity and tumor development, indicating a fine tuning mechanism of p53 activity in vivo. Importantly, each mutant mouse has a distinct phenotype, suggesting diverse and exquisite mechanisms of p53 regulation in different environments, different tissues and different genetic backgrounds. The generation of these mutant p53 knock-in mice has laid the groundwork for future studies to elucidate the in vivo physiological function of mutant p53 and to examine cooperating effects in combination with other alterations. PMID- 17401427 TI - Interactions of mutant p53 with DNA: guilt by association. AB - Since the very early days of p53 research, the gain of oncogenic activities by some mutant p53 proteins had been suspected as an important factor contributing to cancer progression. Considerable progress towards understanding the biology of mutant p53 has been made during the last years, the quintessence being the realization that the impact of mutant p53 proteins on the transcriptome of a tumor cell is much more global than previously thought. The emerging role of mutant p53 proteins in coordinating oncogenic signaling and chromatin modifying activities reveals an until now unsuspected function of these proteins as important modifiers of the oncogenic transcriptional response. Notwithstanding the fact that the sequence-specific DNA binding activity of mutant p53 proteins is impaired, they are still able to associate with specific loci on DNA by utilizing different mechanisms. The ability to associate with DNA appears to be crucial for the master role of mutant p53 proteins in coordinating oncogenic transcriptional responses. PMID- 17401428 TI - Changing the p53 master regulatory network: ELEMENTary, my dear Mr Watson. AB - The p53 master regulatory network provides for the stress-responsive direct control of a vast number of genes in humans that can be grouped into several biological categories including cell-cycle control, apoptosis and DNA repair. Similar to other sequence-specific master regulators, there is a matrix of key components, which provide for variation within the p53 master regulatory network that include p53 itself, target response element sequences (REs) that provide for p53 regulation of target genes, chromatin, accessory proteins and transcription machinery. Changes in any of these can impact the expression of individual genes, groups of genes and the eventual biological responses. The many REs represent the core of the master regulatory network. Since defects or altered expression of p53 are associated with over 50% of all cancers and greater than 90% of p53 mutations are in the sequence-specific DNA-binding domain, it is important to understand the relationship between wild-type or mutant p53 proteins and the target response elements. In the words of the legendary detective Sherlock Holmes, it is 'Elementary, my dear Mr. Watson'. PMID- 17401429 TI - Transcription regulation by mutant p53. AB - In addition to the loss of wild-type p53 activity, a high percentage of tumor cells accumulate mutant p53 protein isoforms. Whereas the hallmark of the wild type p53 is its tumor suppressor activities, tumor-associated mutant p53 proteins acquire novel functions enabling them to promote a large spectrum of cancer phenotypes. During the last years, it became clear that tumor-associated mutant p53 proteins are not only distinct from the wild-type p53, but they also represent a heterogeneous population of proteins with a variety of structure function features. One of the major mechanisms underlying mutant p53 gain of function is the ability to regulate gene expression. Although a large number of specific target genes were identified, the molecular basis for this regulation is not fully elucidated. This review describes the present knowledge about the transcriptional activities of mutant p53 and the mechanisms that might underlie its target gene specificity. PMID- 17401430 TI - Mutant p53: an oncogenic transcription factor. AB - Inactivation of tumor-suppressor genes is one of the key hallmarks of a tumor. Unlike other tumor-suppressor genes, p53 is inactivated by missense mutations in half of all human cancers. It has become increasingly clear that the resulting mutant p53 proteins do not represent only the mere loss of wild-type p53 tumor suppressor activity, but gain new oncogenic properties favoring the insurgence, the maintenance, the spreading and the chemoresistance of malignant tumors. The actual challenge is the fine deciphering of the molecular mechanisms underlying the gain of function of mutant p53 proteins. In this review, we will focus mainly on the transcriptional activity of mutant p53 proteins as one of the potential molecular mechanisms. To date, the related knowledge is still quite scarce and many of the raised questions of this review are yet unanswered. PMID- 17401431 TI - Are interactions with p63 and p73 involved in mutant p53 gain of oncogenic function? AB - Although still controversial, the presence of mutant p53 in cancer cells may result in more aggressive tumors and correspondingly worse outcomes. The means by which mutant p53 exerts such pro-oncogenic activity are currently under extensive investigation and different models have been proposed. We focus here on a proposed mechanism by which a subset of tumor-derived p53 mutants physically interact with p53 family members, p63 and p73, and negatively regulate their proapoptotic function. Both cell-based assays and knock-in mice expressing mutant forms of p53 support this model. As more than half of human tumors harbor mutant forms of p53 protein, approaches aimed at disrupting the pathological interactions among p53 family members might be of clinical value. PMID- 17401432 TI - Structure-function-rescue: the diverse nature of common p53 cancer mutants. AB - The tumor suppressor protein p53 is inactivated by mutation in about half of all human cancers. Most mutations are located in the DNA-binding domain of the protein. It is, therefore, important to understand the structure of p53 and how it responds to mutation, so as to predict the phenotypic response and cancer prognosis. In this review, we present recent structural and systematic functional data that elucidate the molecular basis of how p53 is inactivated by different types of cancer mutation. Intriguingly, common cancer mutants exhibit a variety of distinct local structural changes, while the overall structural scaffold is largely preserved. The diverse structural and energetic response to mutation determines: (i) the folding state of a particular mutant under physiological conditions; (ii) its affinity for the various p53 target DNA sequences; and (iii) its protein-protein interactions both within the p53 tetramer and with a multitude of regulatory proteins. Further, the structural details of individual mutants provide the basis for the design of specific and generic drugs for cancer therapy purposes. In combination with studies on second-site suppressor mutations, it appears that some mutants are ideal rescue candidates, whereas for others simple pharmacological rescue by small molecule drugs may not be successful. PMID- 17401433 TI - Reactivation of mutant p53: molecular mechanisms and therapeutic potential. AB - The p53 tumor suppressor gene is the most frequently mutated gene in cancer. Most p53 mutations are missense point mutations that cluster in the DNA-binding core domain. This results in distortion of core domain folding and disruption of DNA binding and transcriptional transactivation of p53 target genes. Structural studies have demonstrated that mutant p53 core domain unfolding is not irreversible. Mutant p53 is expressed at high levels in many tumors. Therefore, mutant p53 is a promising target for novel cancer therapy. Mutant p53 reactivation will restore p53-dependent apoptosis, resulting in efficient removal of tumor cells. A number of strategies for targeting mutant p53 have been designed, including peptides and small molecules that restore the active conformation and DNA binding to mutant p53 and induce p53-dependent suppression of tumor cell growth in vitro and in vivo. This opens possibilities for the clinical application of mutant p53 reactivation in the treatment of cancer. PMID- 17401434 TI - Why microscopy will remain a cornerstone of surgical pathology. AB - Recent years have seen increasing predictions of the demise of conventional microscopy in patient care and investigative medicine. However, these predictions fail to recognize the power of morphologic analysis by a skilled observer. The amount of information that can be obtained from a simple H&E slide represents a windfall in terms of data quality, quantity and cost when compared to any other available technique. Moreover, the value of such interpretation is irreplaceable as we develop newer and more sophisticated technologies. Overall, it appears that reports of the death of microscopy have been greatly exaggerated. PMID- 17401435 TI - 1,25(OH)(2)D(3) blocks TNF-induced monocytic tissue factor expression by inhibition of transcription factors AP-1 and NF-kappaB. AB - An essential coagulation factor, tissue factor (TF), is rapidly expressed by human monocytes when exposed to a variety of agonists, such as lipopolysaccharide or tumor necrosis factor (TNF). We previously found that 1alpha,25 dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) and its potent synthetic analogs downregulate TF and upregulate thrombomodulin expression on monocytic cells, counteracting the effects of TNF at the level of transcription. The human TF gene has characteristic binding sequences for activator protein-1 (AP-1) (c-Jun/c Fos), nuclear factor-kappaB (NF-kappaB), Sp-1, and early growth response factor-1 (Egr-1). In this study, we investigated the regulatory mechanisms by which 1,25(OH)(2)D(3) inhibits TNF-induced TF expression in human monocytic cells. 1,25(OH)(2)D(3) reduced basal and TNF-induced TF activities. Gel-shift assay and luciferase assay with the respective reporter vectors showed that 1,25(OH)(2)D(3) reduced basal and TNF-induced activities of the nuclear proteins AP-1 and NF kappaB, but not Egr-1. 1,25(OH)(2)D(3) inhibited TNF-induced phosphorylation of c Jun without affecting phosphorylation of the other pathways. On the other hand, 1,25(OH)(2)D(3) directly inhibited nuclear binding and activities of NF-kappaB in the nucleus without affecting phosphorylation of the NF-kappaB activation pathway. These results indicate that 1,25(OH)(2)D(3) suppresses basal and TNF induced TF expression in monocytic cells by inhibition of AP-1 and NF-kappaB activation pathways, but not of Egr-1. Our results may help to elucidate the regulatory mechanisms of 1,25(OH)(2)D(3) in TF induction, and may have physiological significance in the clinical challenge to use potential 1,25(OH)(2)D(3) analogs in antithrombotic therapy as well as immunomodulation and antineoplastic therapy of leukemia. PMID- 17401436 TI - Renal p38 MAP kinase activity in experimental diabetes. AB - Renal cell activity of p38 mitogen-activated protein kinase (p38) is increased in the diabetic milieu. p38 mediates signals relevant for the development of diabetic nephropathy (DN). However, renal p38 in Type 1 diabetes in vivo, particularly in conditions reflecting the differences in metabolic control, and its activity in advanced stages of DN, has received less attention. We examined the p38 pathway in renal cortex of rats with streptozotocin diabetes (4 weeks) with poor (DS), moderate (DM), and intensive (DII) metabolic control, achieved by varying doses of insulin therapy. Renal p38 was also studied in 12-month diabetic rats with established nephropathy (DM12) and compared with age-matched controls. p38 activity (in vitro kinase assay and expression of phosphorylated (active) p38 (P-p38)) was increased in DM and DS rats, as compared with non-diabetic controls, and attenuated by intensive insulin treatment. In all groups, P-p38 was predominantly localized in macula densa cells. Diabetic rats also demonstrated P p38 immunoreactivity in the distal tubule and glomeruli. Enhanced p38 activity in DS and DM rats was not associated with increases in expression of active mitogen activated protein kinase 3/6, an activator of p38, but paralleled with increased expression of scaffolding protein transforming growth factor-beta-activated protein kinase 1-binding protein 1. Expression of mitogen-activated protein phosphatase-1 (MKP-1), one of the phosphatases involved in inactivation of mitogen-activated protein kinase signaling, was increased in all diabetic groups, irrespective of metabolic control. Renal p38 activation was also detectable in D12 rats with established albuminuria and glomerulosclerosis. In summary, renal cortical p38 activity was increased in diabetic rats at early and advanced stages of nephropathy, as compared with non-diabetic animals, and attenuated by improved metabolic control. p38 activation in diabetes is likely to occur via multiple pathways and cannot be explained by downregulation of MKP-1. PMID- 17401437 TI - NO counterbalances HO-1 overexpression-induced acceleration of hepatocyte proliferation in mice. AB - The trigger for liver regeneration, including shear stress, has been the subject of ongoing debate. Blood vessel-derived gaseous molecules carbon monoxide (CO) and nitric oxide (NO) regulate vascular tone and play an important role in liver regeneration. In heme oxygenase-1 (HO-1) transgenic mice, it has been shown that CO-mediated impairment of vasorelaxation is an NO-dependent event. We therefore studied liver regeneration in HO-1 overexpressing animals in dependency of NO availability. Mice were subjected to (2/3) hepatectomy and were treated with either cobalt protoporphyrin-IX for induction of CO-liberating HO-1, N(omega) nitro-L-arginine methyl ester (L-NAME) for blockade of NO synthase (NOS) or both. Application of molsidomine in L-NAME treated animals served for resubstitution of NO. Vehicle-treated animals served as respective control animals. We examined 5 bromo-2'-deoxyuridine incorporation and proliferating cell nuclear antigen expression as well as HO-1 and NOS-2 protein levels. Intrahepatic red blood cell velocity and volumetric blood flow were evaluated by in vivo fluorescence microscopy as indicators for microvascular shear stress. Hepatic regeneration remained unaffected by L-NAME application for NOS blockade. However, NOS blockade in HO-1 induced animals caused increased 5-bromo-2'-deoxyuridine and proliferating cell nuclear antigen measures of liver regeneration. In parallel, these animals revealed increased velocities and volumetric blood flow in the terminal afferent vessels and postsinusoidal venules. These local hemodynamic changes including enhanced hepatocyte proliferation could be reversed by NO liberation via molsidomine. The present findings stress the role of NO to counterbalance vascular tone in HO-1 overexpressing animals for maintenance of adequate perfusion and salutary shear force within the hepatic microvasculature upon liver resection. PMID- 17401438 TI - Histamine H3-receptor agonists and imidazole-based H3-receptor antagonists can be thermodynamically discriminated. AB - BACKGROUND AND PURPOSE: Studies suggest that measurement of thermodynamic parameters can allow discrimination of agonists and antagonists. Here we investigate whether agonists and antagonists can be thermodynamically discriminated at histamine H(3) receptors. EXPERIMENTAL APPROACH: The pK(L) of the antagonist radioligand, [(3)H]-clobenpropit, in guinea-pig cortex membranes was estimated at 4, 12, 21 and 30 degrees C in 20 mM HEPES-NaOH buffer (buffer A), or buffer A containing 300 mM CaCl(2), (buffer A(Ca)). pK(I)' values for ligands with varying intrinsic activity were determined in buffer A and A(Ca) at 4, 12, 21 and 30 degrees C. KEY RESULTS: In buffer A, the pK(L) of [(3)H] clobenpropit increased with decreasing temperature while it did not change in buffer A(Ca). The Bmax was not affected by temperature or buffer and n (H) values were not different from unity. In buffer A, pK(I)' values for agonists remained unchanged or decreased with decreasing temperature, while antagonist pK(I) values increased with decreasing temperature; agonist binding was entropy-driven while antagonist binding was enthalpy and entropy-driven. In buffer A(Ca), temperature had no effect on antagonist and agonist pK(I) values; both agonist and antagonist binding were enthalpy and entropy-driven. CONCLUSIONS AND IMPLICATIONS: The binding of H(3)-receptor agonists and antagonists can be thermodynamically discriminated under conditions where agonist pK(I)' values are over-estimated (pK(I)' not = pK(app)). However, under conditions when agonist pK(I) approximately pK(app), the thermodynamics underlying the binding of agonists are not different to those of antagonists. PMID- 17401439 TI - L-citrulline inhibits [3H]acetylcholine release from rat motor nerve terminals by increasing adenosine outflow and activation of A1 receptors. AB - BACKGROUND AND PURPOSE: Nitric oxide (NO) production and depression of neuromuscular transmission are closely related, but little is known about the role of L-citrulline, a co-product of NO biosynthesis, on neurotransmitter release. EXPERIMENTAL APPROACH: Muscle tension recordings and outflow experiments were performed on rat phrenic nerve-hemidiaphragm preparations stimulated electrically. KEY RESULTS: L-citrulline concentration-dependently inhibited evoked [(3)H]ACh release from motor nerve terminals and depressed nerve-evoked muscle contractions. The NO synthase (NOS) substrate, L-arginine, and the NO donor, 3-morpholinosydnonimine chloride (SIN-1), also inhibited [(3)H]ACh release with a potency order of SIN-1>L-arginine>L-citrulline. Co-application of L citrulline and SIN-1 caused additive effects. NOS inactivation with N(omega) nitro-L-arginine prevented L-arginine inhibition, but not that of L-citrulline. The NO scavenger, haemoglobin, abolished inhibition of [(3)H]ACh release caused by SIN-1, but not that caused by L-arginine. Inactivation of guanylyl cyclase with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) fully blocked SIN-1 inhibition, but only partially attenuated the effects of L-arginine. Reduction of extracellular adenosine accumulation with adenosine deaminase or with the nucleoside transport inhibitor, S-(p-nitrobenzyl)-6-thioinosine, attenuated the effects of L-arginine and L-citrulline, while not affecting inhibition by SIN-1. Similar results were obtained with the selective adenosine A(1) receptor antagonist, 1,3-dipropyl-8-cyclopentylxanthine. L-citrulline increased the resting extracellular concentration of adenosine, without changing that of the adenine nucleotides. CONCLUSIONS AND IMPLICATIONS: NOS catalyses the formation of two neuronally active products, NO and L-citrulline. While, NO may directly reduce transmitter release through stimulation of soluble guanylyl cyclase, the inhibitory action of L-citrulline may be indirect through increasing adenosine outflow and subsequently activating inhibitory A(1) receptors. PMID- 17401440 TI - Cell signaling pathways in the mechanisms of neuroprotection afforded by bergamot essential oil against NMDA-induced cell death in vitro. AB - BACKGROUND AND PURPOSE: The effects of bergamot essential oil (BEO; Citrus bergamia, Risso) on excitotoxic neuronal damage was investigated in vitro. EXPERIMENTAL APPROACH: The study was performed in human SH-SY5Y neuroblastoma cells exposed to N-methyl-D-aspartate (NMDA). Cell viability was measured by dye exclusion. Reactive oxygen species (ROS) and caspase-3 activity were measured fluorimetrically. Calpain I activity and the activation (phosphorylation) of Akt and glycogen synthase kinase-3beta (GSK-3beta) were assayed by Western blotting. KEY RESULTS: NMDA induced concentration-dependent, receptor-mediated, death of SH SY5Y cells, ranging from 11 to 25% (0.25-5 mM). Cell death induced by 1 mM NMDA (21%) was preceded by a significant accumulation of intracellular ROS and by a rapid activation of the calcium-activated protease calpain I. In addition, NMDA caused a rapid deactivation of Akt kinase and this preceded the detrimental activation of the downstream kinase, GSK-3beta. BEO (0.0005-0.01%) concentration dependently reduced death of SH-SY5Y cells caused by 1 mM NMDA. In addition to preventing ROS accumulation and activation of calpain, BEO (0.01%) counteracted the deactivation of Akt and the consequent activation of GSK-3beta, induced by NMDA. Results obtained by using specific fractions of BEO, suggested that monoterpene hydrocarbons were responsible for neuroprotection afforded by BEO against NMDA-induced cell death. CONCLUSIONS AND IMPLICATIONS: Our data demonstrate that BEO reduces neuronal damage caused in vitro by excitotoxic stimuli and that this neuroprotection was associated with prevention of injury induced engagement of critical death pathways. PMID- 17401441 TI - Catecholamine outflow from mouse and rat brain slice preparations evoked by nicotinic acetylcholine receptor activation and electrical field stimulation. AB - BACKGROUND AND PURPOSE: Mice with targeted deletions of neuronal nicotinic acetylcholine receptor (nAChR) subunit genes are valuable models to study nAChR function such as catecholamine outflow by presynaptic receptor activation. Contrary to the rat, our present knowledge on presynaptic nAChRs in mice primarily relies on observations made with synaptosomes. We have now used brain slices to investigate nicotine-induced catecholamine outflow in wild type (WT) and nAChR (beta2 and alpha5) knockout mice for a comparison with rat brain slice preparations. EXPERIMENTAL APPROACH: Brain slices from rat and mouse hippocampus, parieto-occipital neocortex, and corpus striatum were loaded with either [3H] noradrenaline or [3H]-dopamine. We provoked catecholamine outflow by electrical field stimulation and nicotinic agonists. KEY RESULTS: When set in relation to electrical field stimulation, nicotine-evoked catecholamine release was sizeable in the striatum but low in the neocortex of both rats and mice. [3H] noradrenaline outflow was, on the other hand, substantial in the rat but low in the mouse hippocampus. About 10% (or less) of nicotine-induced catecholamine release persisted in the presence of tetrodotoxin in all our preparations. CONCLUSIONS AND IMPLICATIONS: Targeted deletion of the beta2 subunit gene essentially abolished the effect of nicotine, indicating that this subunit is an essential constituent of nAChRs that indirectly (via action potentials) induce catecholamine release from hippocampal and striatal slices in mice. The impact of nAChRs in catecholaminergic projection areas differs between species and has thus to be considered when extrapolating results from animal models to human conditions. PMID- 17401443 TI - Aminoguanidine prevents fructose-induced deterioration in left ventricular arterial coupling in Wistar rats. AB - BACKGROUND AND PURPOSE: Aminoguanidine (AG), an inhibitor of advanced glycation endproducts, has been identified as a prominent agent that prevents the fructose induced arterial stiffening in male Wistar rats. Our aims were to examine whether AG produced benefits on the left ventricular (LV)-arterial coupling in fructose fed (FF) animals in terms of the ventricular and arterial chamber properties. EXPERIMENTAL APPROACH: Rats given 10% fructose in drinking water (FF) were daily treated with AG (50 mg x kg(-1), i.p.) for 2 weeks and compared with the untreated FF group. In anaesthetised rats, LV pressure and ascending aortic flow signals were recorded to calculate LV end-systolic elastance (E(es), an indicator of myocardial contractility) and effective arterial volume elastance (E(a)). The optimal afterload (Q(load)) determined by the ratio of E(a) to E(es) was used to measure the coupling efficiency between the left ventricle and its vasculature. KEY RESULTS: There was a significant interaction between fructose and AG in their effects on E(a). Fructose loading significantly elevated E(a) and AG prevented the fructose-derived deterioration in arterial chamber elastance. Both fructose and AG affected E(es) and Q(load), and there was an interaction between fructose and AG for these two variables. Both E(es) and Q(load) exhibited a decline with fructose feeding but showed a significant rise after AG treatment in the FF rats. CONCLUSIONS AND IMPLICATIONS: AG prevented not only the contractile dysfunction of the heart caused by fructose loading, but also the fructose-induced deterioration in matching left ventricular function to the arterial system. PMID- 17401442 TI - Recent developments in nitric oxide donor drugs. AB - During the 1980s, the free radical, nitric oxide (NO), was discovered to be a crucial signalling molecule, with wide-ranging functions in the cardiovascular, nervous and immune systems. Aside from providing a credible explanation for the actions of organic nitrates and sodium nitroprusside that have long been used in the treatment of angina and hypertensive crises respectively, the discovery generated great hopes for new NO-based treatments for a wide variety of ailments. Decades later, however, we are still awaiting novel licensed agents in this arena, despite an enormous research effort to this end. This review explores some of the most promising recent advances in NO donor drug development and addresses the challenges associated with NO as a therapeutic agent. PMID- 17401445 TI - [Actuality of Semmelweis]. PMID- 17401444 TI - YC-1 attenuates LPS-induced proinflammatory responses and activation of nuclear factor-kappaB in microglia. AB - BACKGROUND AND PURPOSE: An inflammatory response in the central nervous system mediated by the activation of microglia is a key event in the early stages of the development of neurodegenerative diseases. LPS has been reported to cause marked microglia activation. It is very important to develop drugs that can inhibit microglia activation and neuroinflammation. Here, we investigated the inhibitory effect of YC-1, a known activator of soluble guanylyl cyclase, against LPS induced inflammatory responses in microglia. EXPERIMENTAL APPROACH: To understand the inhibitory effects of YC-1 on LPS-induced neuroinflammation, primary cultures of rat microglia and the microglia cell line BV-2 were used. To examine the mechanism of action of YC-1, LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production, iNOS, COX-2 and cytokine expression were analyzed by Griess reaction, ELISA, Western blotting and RT-PCR, respectively. The effect of YC-1 on LPS-induced activation of nuclear factor kappa B (NF-kappaB) was studied by NF kappaB reporter assay and immunofluorocytochemistry. KEY RESULTS: YC-1 inhibited LPS-induced production of NO and PGE2 in a concentration-dependent manner. The protein and mRNA expression of iNOS and COX-2 in response to LPS application were also decreased by YC-1. In addition, YC-1 effectively reduced LPS-induced expression of the mRNA for the proinflammatory cytokines, TNF-alpha and IL-1beta. Furthermore, YC-1 inhibited LPS-induced NF-kappaB activation in microglia. CONCLUSIONS AND IMPLICATIONS: YC-1 was able to inhibit LPS-induced iNOS and COX-2 expression and NF-kappaB activation, indicating that YC-1 may be developed as an anti-inflammatory neuroprotective agent. PMID- 17401446 TI - [A child health promotion intervention in Albania: results and lessons learned]. AB - Albania is a Balkan country in South-Eastern Europe which, in recent years, has undergone complex demographic, political and economical changes. A notable drop in infant and maternal mortality rates and a significant rise in economic indicators have been observed in recent years. Despite this, over 15% of the population living in the northern and north-eastern areas of the country lives in extreme poverty conditions. In recent years various healthcare system reforms have been introduced, including the introduction of private healthcare and improvement of the main hospital infrastructures but not much has been done to increase the provision of essential healthcare services especially in rural and poor areas. Inequalities in health care are therefore widespread and these particularly affect children living in critical areas. In this paper we describe a paediatric healthcare intervention programme conducted in Albania from 2002 to 2004, aimed at improving the health and nutrition status of children and tackling healthcare system inequalities. The intervention consisted in offering free healthcare services and assistance, delivered through the Albanian healthcare system, to 5280 children. It also involved a health education programme for the mothers. The impact of the programme on the prevalence of infant malnutrition was evaluated by examining the medical records of 1745 infants followed for at least 6 months. Prevalence of malnutrition significantly decreased, from 13.4% to 4.2% during the study period. Mortality in children aged 0-5 years also showed a considerable drop. These results confirm that an efficient and sustainable model of paediatric healthcare assistance in Albania is possible. PMID- 17401447 TI - [Microbiological quality of haemodialysis water in various hospitals and private clinics in the Lazio region (Italy): 2000-2004]. AB - The aim of this study was to evaluate the quality of dialysis water used in dialysis units of various hospitals and private clinics in Rome and the Lazio region (Italy). The study was performed between January 2000 and December 2004. A total of 507 samples were analysed, 205 of which obtained from public hospitals and 302 from privately-owned medical facilities. Microbiological analysis was performed as specified by the 1995 Decree of the Lazio Region n degrees 1650, which also sets threshold values for bacterial contamination and minimum frequency of inspections. The decree also specifies the modalities and frequency of disinfection of dialysis water treatment and distribution systems. The findings show a superior microbiological quality of dialysis water in privately owned dialysis units. This may be due to several factors but especially to a more regular maintenance and disinfection of the water treatment systems. Overall, over half of the samples analysed, in both types of facilities, showed bacterial loads well below the set limits. In our view, however, bacterial limits for dialysis water are too high especially considering the fact that they are the same limits set by previous regulations for drinkable water. Haemodialysis patients are exposed to large volumes of water used for the production of dialysis fluids during a treatment session. It is therefore clearly important that more stringent limits for bacterial contamination be introduced. PMID- 17401448 TI - [A retrospective study to evaluate the incidence of medication errors occurring in the prescription and transcription steps of the medication use process]. AB - Medication errors occur frequently in many clinical settings. Various studies have highlighted that, together with adverse drug events, they represent one of the major causes of adverse events occurring in hospitals. The aim of this study was to perform a detailed retrospective medical record review in order to investigate the incidence of medication errors occurring in the prescription and transcription phases of the medication use process. Overall, 56 medical records were reviewed to determine the incidence of incomplete or incorrect prescriptions and incorrect transcription by nurses of the original medication order. The findings highlight the need to improve medication safety, in particular at the time of prescription and transcription of orders. The study also confirms that the retrospective review of medical records is an effective method for identifying certain types of medication errors that occur during the prescription and transcription phases. However, this type of review is complex and too costly to be used routinely, for continuous monitoring, in clinical practice. PMID- 17401449 TI - [Injury-related Emergency Department visits in Tuscany (Italy)]. AB - The frequency of injuries underscores the need for planning and implementing efficient injury surveillance systems. Emergency Departments represent the preferred source of data on injuries but information regarding emergency department visits is not always available. We examined the feasibility of utilizing emergency department data to monitor the occurrence of injuries in Tuscany. Each of the 52 public hospital emergency departments operating in the 12 local health units of Tuscany were asked to provide yearly data on the numbers and types of injury-related visits since the year 2003. They were also asked whether attendance records were computerised. This data was used to estimate the number of injury-related visits by cause of injury, at the regional level. The results of this study were combined with those of a previous study to estimate the number of hospital admissions for motor vehicle accidents in Tuscany in 2004. The latter was then compared to corresponding data from the hospital discharge abstract database. In 2002, the number of emergency departments with computerised attendance records was only 27 while in 2005 it was 43 and a greater number of emergency departments were able to codify each type of injury-related visit. A slight decrease was observed in the total number of visits for all causes, between 2002 and 2004 (respectively 1.314.874 and 1.256.509). In 2002, motor vehicle accidents were the most frequent cause of injury-related ED visit (8%), followed by home injuries (7.2%) and workplace accidents (6%). In 2004, home injuries were the most frequent type of injury (7.5%) followed by motor vehicle accidents (6.9%) and workplace injuries (5%). We estimated 6836 hospital admissions for the year 2004, while only 4800 admissions were registered in the discharge abstract database in the same year. Computerisation of attendance records and improvement in codification of data are a good starting point in utilizing emergency department data for epidemiological surveillance of injuries. PMID- 17401450 TI - [Strategies of immunization in Ghana: the experience of a "community-based" health planning in a rural country]. AB - Access to immunization of children and to prevention services is a relevant issue in poor-resource settings like rural areas of Western Africa. Ghana government has launched the "Community-based Health Planning and Services initiative-CHPS", a programme that by the institution of Local Clinics in small villages, the activity of their nurse, and the involvement of local communities and traditional institutions improves the access of population to primary care and prevention. Our survey in Jomoro district has confirmed that this model is effective in determining higher coverage rate for all the immunizations of the children. PMID- 17401451 TI - Positive feedback may cause the biphasic response observed in the chemoattractant induced response of Dictyostelium cells. AB - After stimulation by chemoattractant, Dictyostelium cells exhibit a rapid response. The concentrations of several intracellular proteins rise rapidly reaching their maximum levels approximately 5-10 seconds, after which they return to prestimulus levels. This response, which is found in many other chemotaxing cells, is an example of a step disturbance rejection, a process known to biologists as perfect adaptation. Unlike other cells, however, the initial first peak observed in the chemoattractant-induced response of Dictyostelium cells is then followed by a slower, smaller phase peaking approximately one to two minutes after the stimulus. Until recently, the nature of this biphasic response has been poorly understood. Moreover, the origin for the second phase is unknown. In this paper we conjecture the existence of a feedback path between the response and stimulus. Using a mathematical model of the chemoattractant-induced response in cells, and standard tools from control engineering, we show that positive feedback may elicit this second peak. PMID- 17401452 TI - Numerical Approximation of Solutions of a Nonlinear Inverse Problem Arising in Olfaction Experimentation. AB - Identification of detailed features of neuronal systems is an important challenge in the biosciences today. Cilia are long thin structures that extend from the olfactory receptor neurons into the nasal mucus. Transduction of an odor into an electrical signal occurs in the membranes of the cilia. The cyclic-nucleotide gated (CNG) channels which reside in the ciliary membrane and are activated by adenosine 3',5'-cyclic monophosphate (cAMP) allow a depolarizing influx of Ca(2+) and Na(+) and are thought to initiate the electrical signal.In this paper, a mathematical model consisting of two nonlinear differential equations and a constrained Fredholm integral equation of the first kind is developed to model experiments involving the diffusion of cAMP into cilia and the resulting electrical activity. The unknowns in the problem are the concentration of cAMP, the membrane potential and, the quantity of most interest in this work, the distribution of CNG channels along the length of a cilium. A simple numerical method is derived that can be used to obtain estimates of the spatial distribution of CNG ion channels along the length of a cilium. Certain computations indicate that this mathematical problem is ill-conditioned. PMID- 17401453 TI - Facial Expressions of Emotion: Are Angry Faces Detected More Efficiently? AB - The rapid detection of facial expressions of anger or threat has obvious adaptive value. In this study, we examined the efficiency of facial processing by means of a visual search task. Participants searched displays of schematic faces and were required to determine whether the faces displayed were all the same or whether one was different. Four main results were found: (1) When displays contained the same faces, people were slower in detecting the absence of a discrepant face when the faces displayed angry (or sad/angry) rather than happy expressions. (2) When displays contained a discrepant face people were faster in detecting this when the discrepant face displayed an angry rather than a happy expression. (3) Neither of these patterns for same and different displays was apparent when face displays were inverted, or when just the mouth was presented in isolation. (4) The search slopes for angry targets were significantly lower than for happy targets. These results suggest that detection of angry facial expressions is fast and efficient, although does not "pop-out" in the traditional sense. PMID- 17401454 TI - Much ado about nothing: A comparison of missing data methods and software to fit incomplete data regression models. AB - Missing data are a recurring problem that can cause bias or lead to inefficient analyses. Development of statistical methods to address missingness have been actively pursued in recent years, including imputation, likelihood and weighting approaches. Each approach is more complicated when there are many patterns of missing values, or when both categorical and continuous random variables are involved. Implementations of routines to incorporate observations with incomplete variables in regression models are now widely available. We review these routines in the context of a motivating example from a large health services research dataset. While there are still limitations to the current implementations, and additional efforts are required of the analyst, it is feasible to incorporate partially observed values, and these methods should be utilized in practice. PMID- 17401455 TI - Evidence-based treatment for HIV-associated dementia and cognitive impairment: why so little? PMID- 17401456 TI - Factors in AIDS dementia complex trial design: results and lessons from the abacavir trial. AB - OBJECTIVES: To determine the efficacy of adding abacavir (Ziagen, ABC) to optimal stable background antiretroviral therapy (SBG) to AIDS dementia complex (ADC) patients and address trial design. DESIGN: Phase III randomized, double-blind placebo-controlled trial. SETTING: Tertiary outpatient clinics. PARTICIPANTS: ADC patients on SBG for > or = 8 wk. INTERVENTIONS: Participants were randomized to ABC or matched placebo for 12 wk. OUTCOME MEASURES: The primary outcome measure was the change in the summary neuropsychological Z score (NPZ). Secondary measures were HIV RNA and the immune activation markers beta-2 microglobulin, soluble tumor necrosis factor (TNF) receptor 2, and quinolinic acid. RESULTS: 105 participants were enrolled. The median change in NPZ at week 12 was +0.76 for the ABC + SBG and +0.63 for the SBG groups (p = 0.735). The lack of efficacy was unlikely related to possible limited antiviral efficacy of ABC: at week 12 more ABC than placebo participants had plasma HIV RNA < or = 400 copies/mL (p = 0.002). There were, however, other factors. Two thirds of patients were subsequently found to have had baseline resistance to ABC. Second, there was an unanticipated beneficial effect of SBG that extended beyond 8 wk to 5 mo, thereby rendering some of the patients at baseline unstable. Third, there was an unexpectedly large variability in neuropsychological performance that underpowered the study. Fourth, there was a relative lack of activity of ADC: 56% of all patients had baseline cerebrospinal fluid (CSF) HIV-1 RNA < 100 copies/mL and 83% had CSF beta-2 microglobulin < 3 nmol/L at baseline. CONCLUSIONS: The addition of ABC to SBG for ADC patients was not efficacious, possibly because of the inefficacy of ABC per se, baseline drug resistance, prolonged benefit from existing therapy, difficulties with sample size calculations, and lack of disease activity. Assessment of these trial design factors is critical in the design of future ADC trials. PMID- 17401457 TI - Low expression of ARHI is associated with shorter progression-free survival in pancreatic endocrine tumors. AB - Little is known about the molecular anomalies involved in the development and progression of malignancy of pancreatic endocrine tumors (PETs). A recently identified member of the Ras family, Ras homologue member I (ARHI), has been shown to be involved in breast, ovary, and thyroid carcinogenesis. Unlike other members, it acts as a tumor suppressor gene that inhibits cell growth. Here we analyzed the mRNA expression of ARHI in 52 primary PETs and 16 normal pancreata using quantitative reverse transcription-polymerase chain reaction. ARHI expression showed a statistically significant difference between either normal pancreas or well-differentiated endocrine tumors (WDET) and poorly differentiated endocrine carcinomas (PDECs) (P < .001 and P < .001, respectively). Moreover, ARHI expression among WDEC samples was more heterogeneous than in WDET, with several tumors showing level of expression analogous to that observed in PDECs. A significant correlation between lower ARHI expression and shorter survival (P = .020) was identified, and a low ARHI expression was associated to a shorter time to progression (P < .001), even considering the proliferation index Ki67 in the multivariate analysis. ARHI is involved in PET progression. Its mRNA expression seemed to be a prognostic factor for disease outcome and, in association with the proliferative index Ki67, a predictor for a rapid tumor relapse. PMID- 17401458 TI - Ets1 transcription factor mediates gastrin-releasing peptide-induced IL-8 regulation in neuroblastoma cells. AB - Angiogenesis plays a critical role in tumor progression in various cancers, including neuroblastoma. We have previously shown that gastrin-releasing peptide (GRP) stimulates neuroblastoma growth and that its cell surface receptors, gastrin-releasing peptide receptors (GRP-R), are overexpressed in advanced-stage human neuroblastomas; however, the effects of GRP on angiogenesis are not clearly elucidated. Interleukin (IL) 8, a proinflammatory chemokine, plays an important role during tumor angiogenesis. Ets transcription factors, such as oncoproteins, cause tumor development and are also known to induce IL-8 expression. In the present study, we found an increased expression of Ets1 in more undifferentiated human neuroblastomas. Stable transfection of SK-N-SH human neuroblastoma cells with Ets1 plasmid resulted in increased IL-8 luciferase activity and IL-8 secretion into cell culture media. Conversely, silencing of Ets1 resulted in a significant decrease in IL-8 secretion in SK-N-SH cells. Moreover, exogenous GRP treatment increased Ets1 (T38) phosphorylation and Ets1 nuclear accumulation, and enhanced Ets1 binding to its DNA consensus sequence, resulting in the stimulation of IL-8 mRNA expression and protein secretion. Our findings demonstrate that GRP upregulates proangiogenic IL-8 expression in an Ets1-dependent manner, suggesting a critical role of this process during GRP-induced neuroblastoma angiogenesis and metastasis. PMID- 17401459 TI - Sulindac induces apoptosis and inhibits tumor growth in vivo in head and neck squamous cell carcinoma. AB - Sulindac has antineoplastic effects on various cancer cell lines; consequently, we assessed sulindac's effects on laryngeal squamous cell carcinoma (SCC) cells in vitro and in vivo. In vitro, SCC (HEP-2) cells treated with various cyclooxygenase inhibitors or transfected with constitutively active signal transducer and activator of transcription 3 (Stat3) or survivin vectors were analyzed using Western blot analysis, annexin V assay, and cell proliferation assay. In parallel, nude mice injected subcutaneously with HEP-2 cells were either treated intraperitoneally with sulindac or left untreated, and analyzed for tumor weight, survivin expression, and tyrosine-phosphorylated Stat3 expression. In vitro studies confirmed the selective antiproliferative and proapoptotic effects of sulindac, which also downregulated Stat3 and survivin protein expression. Stat3 or survivin forced expression partially rescued the antiproliferative effects of sulindac. In vivo studies showed significant repression of HEP-2 xenograft growth in sulindactreated mice versus controls, with near-complete resolution at 10 days. Additionally, tumor specimens treated with sulindac showed downregulation of phosphorylated tyrosine-705 Stat3 and survivin expression. Taken together, our data suggest, for the first time, a specific inhibitory effect of sulindac on tumor growth and survivin expression in laryngeal cancer, both in vitro and in vivo, in a Stat3-dependent manner, suggesting a novel therapeutic approach to head and neck cancer. PMID- 17401460 TI - Molecular characterization of TMPRSS2-ERG gene fusion in the NCI-H660 prostate cancer cell line: a new perspective for an old model. AB - Recent studies have established that a significant fraction of prostate cancers harbor a signature gene fusion between the 5' region of androgen-regulated TMPRSS2 and an ETS family transcription factor, most commonly ERG. Studies on the molecular mechanisms and functional consequences of this important chromosomal rearrangement are currently limited to the VCaP cell line derived from a vertebral bone metastasis of a hormone-refractory prostate tumor. Here we report on the NCI-H660 cell line, derived from a metastatic site of an extrapulmonary small cell carcinoma arising from the prostate. NCI-H660 harbors TMPRSS2-ERG fusion with a homozygous intronic deletion between TMPRSS2 and ERG. We demonstrate this by real-time quantitative polymerase chain reaction, a two-stage dual-color interphase fluorescence in situ hybridization (FISH) assay testing for TMPRSS2 and ERG break-aparts, and single-nucleotide polymorphism oligonucleotide arrays. The deletion is consistent with the common intronic deletion found on chromosome 21q22.2-3 in human prostate cancer samples. We demonstrate the physical juxtaposition of TMPRSS2 and ERG on the DNA level by fiber FISH. The androgen receptor-negative NCI-H660 cell line expresses ERG in an androgen independent fashion. This in vitro model system has the potential to provide important pathobiologic insights into TMPRSS2-ERG fusion prostate cancer. PMID- 17401461 TI - Gene expression profiling of chemically induced rat bladder tumors. AB - A variety of genetic alterations and gene expression changes are involved in the pathogenesis of bladder tumors. To explore expression changes in 4 hydroxybutyl(butyl)nitrosamine-induced rat bladder tumors, microarray analysis was performed. Analysis yielded 1,138 known genes and 867 expressed sequence tags that were changed when comparing tumors to normal rat epithelia. Altered genes included cell cycle-related genes, EGFR-Ras signaling genes, apoptosis genes, growth factors, and oncogenes. Using the pathway visualization tool GenMAPP, we found that these genes can be grouped along several pathways that control apoptosis, cell cycle, and integrin-mediated cell adhesion. When comparing current data with previous mouse bladder tumor data, we found that > 280 of the same known genes were differentially expressed in both mouse and rat bladder tumors, including cell cycle-related genes, small G proteins, apoptosis genes, oncogenes, tumor-suppressor genes, and growth factors. These results suggest that multiple pathways are involved in rat bladder tumorigenesis, and a common molecular mechanism was found in both rat and mouse bladder tumors. PMID- 17401462 TI - Anti-inflammatory agent indomethacin reduces invasion and alters metabolism in a human breast cancer cell line. AB - Hostile physiological environments such as hypoxia and acidic extracellular pH, which exist in solid tumors, may promote invasion and metastasis through inflammatory responses and formation of eicosanoids. Here, we have investigated the effects of the anti-inflammatory agent indomethacin on the invasion and metabolism of the human breast cancer cell line MDA-MB-435 in Dulbecco's Modified Eagles (DME)-based or Roswell Park Memorial Institute (RPMI)-based cell medium, using a magnetic resonance-compatible invasion assay. Indomethacin treatment significantly reduced the invasion of MDA-MB-435 cells independent of the culture and perfusion conditions examined. Significant changes were detected in levels of intracellular choline phospholipid metabolites and in triglyceride (TG) concentrations of these cells, depending on indomethacin treatment and basal cell medium used. Additionally, genetic profiling of breast cancer cells, grown and treated with low-dose indomethacin in cell culture using an RPMI-based medium, revealed the upregulation of several genes implicating cyclooxygenase-independent targets of indomethacin. These data confirm the ability of an anti-inflammatory agent to reduce breast cancer invasion and demonstrate, depending on cell culture and perfusion conditions, that the indomethacin-induced decrease in invasion is associated with changes in choline phospholipid metabolism, TG metabolism, and gene expression. PMID- 17401463 TI - Early involvement of death-associated protein kinase promoter hypermethylation in the carcinogenesis of Barrett's esophageal adenocarcinoma and its association with clinical progression. AB - Esophageal Barrett's adenocarcinoma (BA) develops through a multistage process, which is associated with the transcriptional silencing of tumor-suppressor genes by promoter CpG island hypermethylation. In this study, we explored the promoter hypermethylation and protein expression of proapoptotic death-associated protein kinase (DAPK) during the multistep Barrett's carcinogenesis cascade. Early BA and paired samples of premalignant lesions of 61 patients were analyzed by methylation-specific polymerase chain reaction and immunohistochemistry. For the association of clinicopathological markers and protein expression, an immunohistochemical tissue microarray analysis of 66 additional BAs of advanced tumor stages was performed. Hypermethylation of DAPK promoter was detected in 20% of normal mucosa, 50% of Barrett's metaplasia, 53% of dysplasia, and 60% of adenocarcinomas, and resulted in a marked decrease in DAPK protein expression (P < .01). The loss of DAPK protein was significantly associated with advanced depth of tumor invasion and advanced tumor stages (P < .001). Moreover, the severity of reflux esophagitis correlated significantly with the hypermethylation rate of the DAPK promoter (P < .003). Thus, we consider DAPK inactivation by promoter hypermethylation as an early event in Barrett's carcinogenesis and suggest that a decreased protein expression of DAPK likely plays a role in the development and progression of BA. PMID- 17401464 TI - Antagonistic effects of sodium butyrate and N-(4-hydroxyphenyl)-retinamide on prostate cancer. AB - Butyrates and retinoids are promising antineoplastic agents. Here we analyzed effects of sodium butyrate and N-(4-hydroxyphenyl)-retinamide (4-HPR) on prostate cancer cells as monotherapy or in combination in vitro and in vivo. Sodium butyrate and 4-HPR induced concentration-dependent growth inhibition in prostate cancer cells in vitro. The isobologram analysis revealed that sodium butyrate and 4-HPR administered together antagonize effects of each other. For the in vivo studies, a water-soluble complex (4-HPR with a cyclodextrin) was created. A single dose of sodium butyrate and 4-HPR showed a peak level in chicken plasma within 30 minutes. Both compounds induced inhibition of proliferation and apoptosis in xenografts of the chicken chorioallantoic membrane. Analysis of the cytotoxic effects of the drugs used in combination demonstrated an antagonistic effect on inhibition of proliferation and on induction of apoptosis. Prolonged jun N-terminal kinase phosphorylation induced by sodium butyrate and 4-HPR was strongly attenuated when both compounds were used in combination. Both compounds induced inhibition of NF-kappaB. This effect was strongly antagonized in LNCaP cells when the compounds were used in combination. These results indicate that combinational therapies have to be carefully investigated due to potential antagonistic effects in the clinical setting despite promising results of a monotherapy. PMID- 17401465 TI - Posttranslational modifications of transthyretin are serum markers in patients with mycosis fungoides. AB - Cutaneous T-cell lymphomas (CTCLs) are characterized by the recruitment of malignant T-cell clones, predominantly of the CD4(+) T-helper subpopulation, into the skin. Mycosis fungoides (MF) is the most common type of CTCL and accounts for almost 50% of all primary cutaneous lymphomas. The ProteinChip technology surface enhanced laser desorption/ionization time of flight/mass spectrometry (SELDI-TOF MS) was used to detect biomarkers in sera from MF patients (n = 25) and healthy controls (n = 26). Therefore, diluted sera were applied to IMAC30 ProteinChip arrays, and the resulting protein profiles were bioinformatically analyzed. A protein set that distinguishes MF patients from healthy controls with a sensitivity of 82.6% and a specificity of 100% was identified. Four significant peaks were identified by two-dimensional gel electrophoresis, immunodepletion, and SELDI-TOF-MS as transthyretin (TTR) and three TTR modifications. A subsequent enzyme-linked immunosorbent assay confirmed these findings. The ability to detect and identify proteins and protein modifications using SELDI-TOF-MS might reveal a better insight on this kind of disease and may lead to a better understanding and earlier detection of MF patients. PMID- 17401466 TI - Prognostic significance of immunoreactive neutrophil elastase in human breast cancer: long-term follow-up results in 313 patients. AB - OBJECTIVE: We have measured the concentration of immunoreactive neutrophil elastase (ir-NE) in the tumor extracts of 313 primary human breast cancers. Sufficient time has elapsed, and we are now ready to analyze its prognostic value in human breast cancer. METHODS: ir-NE concentration in tumor extracts was determined with an enzyme-linked immunosorbent assay that enables a rapid measurement of both free-form ir-NE and the A1-protease inhibitor-complexed form of ir-NE. We analyzed the prognostic value of this enzyme in human breast cancer in univariate and multivariate analyses. RESULTS: Patients with breast cancer tissue containing a high concentration of ir-NE had poor survival compared to those with a low concentration of ir-NE at the cutoff point of 9.0 microg/100 mg protein (P = .0012), which had been previously determined in another group of 49 patients. Multivariate stepwise analysis selected lymph node status (P = .0004; relative risk = 1.46) and ir-NE concentration (P = .0013; relative risk = 1.43) as independent prognostic factors for recurrence. CONCLUSIONS: Tumor ir-NE concentration is an independent prognostic factor in patients with breast cancer who undergo curative surgery. This enzyme may play an active role in tumor progression that leads to metastasis in human breast cancer. PMID- 17401467 TI - Crossover comparison study on the therapeutic effects of tamsulosin hydrochloride and naftopidil in lower urinary tract symptoms associated with benign prostatic hyperplasia. AB - There is no consensus on the evaluated efficacy of individual alpha(1)-blockers in the treatment of lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH). This study compared the clinical benefits of tamsulosin hydrochloride with those of naftopidil in BPH. Men aged 54-84 years with a main complaint of BPH were randomly assigned to receive consecutive treatment with tamsulosin then naftopidil (T-N group, 25 patients), or naftopidil then tamsulosin hydrochloride (N-T group, 20 patients). The therapeutic effects were compared in a crossover design. Administration was continued for 28 days in each treatment period. Both groups showed similar improvements during the first treatment period. However, during the second treatment period after crossover, therapeutic effects were greater in the N-T group. Tamsulosin was more effective than naftopidil on intermittency, nocturia and quality of life scores. Tamsulosin and naftopidil have different efficacy profiles for LUTS associated with BPH. PMID- 17401468 TI - On focal stability in dimension two. AB - In Kupka et al. 2006 appears the Focal Stability Conjecture: the focal decomposition of the generic Riemann structure on a manifold M is stable under perturbations of the Riemann structure. In this paper, we prove the conjecture when M has dimension two, and there are no conjugate points. PMID- 17401469 TI - Finitely curved orbits of complex polynomial vector fields. AB - This note is about the geometry of holomorphic foliations. Let X be a polynomial vector field with isolated singularities on C2. We announce some results regarding two problems: 1. Given a finitely curved orbit L of X, under which conditions is L algebraic? 2. If X has some non-algebraic finitely curved orbit L what is the classification of X? Problem 1 is related to the following question: Let C [symbol: see text]C2 be a holomorphic curve which has finite total Gaussian curvature. Is C contained in an algebraic curve? PMID- 17401470 TI - New tools to study biophysical properties of single molecules and single cells. AB - We present a review on two new tools to study biophysical properties of single molecules and single cells. A laser incident through a high numerical aperture microscope objective can trap small dielectric particles near the focus. This arrangement is named optical tweezers. This technique has the advantage to permit manipulation of a single individual object. We use optical tweezers to measure the entropic elasticity of a single DNA molecule and its interaction with the drug Psoralen. Optical tweezers are also used to hold a kidney cell MDCK away from the substrate to allow precise volume measurements of this single cell during an osmotic shock. This procedure allows us to obtain information about membrane water permeability and regulatory volume increase. Defocusing microscopy is a recent technique invented in our laboratory, which allows the observation of transparent objects, by simply defocusing the microscope in a controlled way. Our physical model of a defocused microscope shows that the image contrast observed in this case is proportional to the defocus distance and to the curvature of the transparent object. Defocusing microscopy is very useful to study motility and mechanical properties of cells. We show here the application of defocusing microscopy to measurements of macrophage surface fluctuations and their influence on phagocytosis. PMID- 17401471 TI - 1,4- Addition of diazomethane to a heterodiene: a direct preparation of the oxazolic ring. AB - The reaction of naphthoquinone-oximes (3) and (4) with diazomethane yields directly, in one step, the oxazoles (5) and (6), respectively. PMID- 17401472 TI - Insecticidal activity of 2-tridecanone against the cowpea weevil Callosobruchus maculatus (Coleoptera: Bruchidae). AB - The effect of 2-tridecanone vapor on the cowpea weevil (Callosobruchus maculatus) development was determined. Seeds of cowpea were infested with adults and exposed to different doses of 2-tridecanone isolated from Pilocarpus microphyllus Stapf ex Holm, a plant species native from northeastern Brazil. The pure monoterpene was evaluated both undiluted as well as in the dilutions 1:10, 1:100 and 1:1,000 (v/v). The following parameters of the cowpea weevil life cycle were analyzed in response to decreasing doses of 2-tridecanone: number of eggs laid, percentage of egg hatching on seeds, percentage of adult emergence, adult weight at emergence, mean developmental time and number of adults emerged. Vapor of 2-tridecanone caused a significant (P < 0.05) reduction in the number of eggs laid, in the percentage of eggs hatched and in the number of emerged adults in infested seeds. The fumigant insecticidal effect of 2-tridecanone was mainly due to its ovicidal activity. PMID- 17401473 TI - Spatial variation in the stable isotopes of 13C and 15N and trophic position of Leporinus friderici (Characiformes, Anostomidae) in Corumba Reservoir, Brazil. AB - Stable isotopes of carbon (delta13C) and nitrogen (delta15N) were used to describe sources of energy and trophic position for adult Leporinus friderici in the area of the Corumba Reservoir, Brazil. Samples were collected from April 1999 to March 2000. Spatial variations were not identified in the isotopic composition. The maximum and minimum contribution of C4 plants calculated integrating the variation of plants and fish were 47.7% and 2.4%, respectively. Among C3 plants, periphyton presented closer isotopic values to those observed for fishes, corresponding to an important carbon source. The proportion of ingested plant item is larger in rivers upstream from the reservoir (42.7%), which justifies the smaller trophic level among there. However, in the reservoir, the ingestion of fish was 81.4%, while ingested plants contributed with 18.6%. Downstream from the dam, participation of plant item was even smaller (14.4%). Although the trophic position calculated with diet data was proportional to the one calculated with delta15N values, the former elevated the trophic level of L. friderici in the food web, because estimated trophic positions were based on fish items belonging to the 2nd (a) and to the 3rd (b) trophic levels. PMID- 17401475 TI - Allosteric regulation of 6-phosphofructo-1-kinase activity of fat body and flight muscle from the bloodsucking bug Rhodnius prolixus. AB - 6-Phosphofructo-1-kinase (phosphofructokinase; PFK) activity from Rhodnius prolixus, a haematophagous insect which is usually a poor flyer, was measured and compared in two metabolically active tissues - flight muscle and fat body. The activity of this important regulatory glycolytic enzyme was much more pronounced in muscle (15.1 +/- 1.4 U/mg) than in fat body extracts (3.6+/-0.4 U/mg), although the latter presented higher levels of enzyme per protein content, as measured by western-blotting. Muscle extracts are more responsible than fat body to ATP and fructose 6-phosphate, both substrates of PFK. Allosteric regulation exerted by different effectors such as ADP, AMP and fructose 2,6-phosphate presented a singular pattern for each tissue. Optimal pH (8.0-8.5) and sensitivity to pH variation was very similar, and citrate was unable to inhibit PFK activity in both extracts. Our results suggest the existence of a particular PFK activity for each tissue, with regulatory patterns that are consistent with their physiological roles. PMID- 17401476 TI - Pore-linked filaments in anura spermatocyte nuclei. AB - Pore-linked filaments were visualized in spreads of anuran spermatocyte nuclei using transmission electron microscope. We used Odontophrynus diplo and tetraploid species having the tetraploid frogs reduced metabolic activities. The filaments with 20-40 nm width are connected to a ring component of the nuclear pore complex with 90-120 nm and extend up to 1 microm (or more) into the nucleus. The filaments are curved and connect single or neighboring pores. The intranuclear filaments are associated with chromatin fibers and related to RNP particles of 20-25 nm and spheroidal structures of 0.5 microm, with variations. The aggregates of several neighboring pores with the filaments are more commonly observed in 4n nuclei. We concluded that the intranuclear filaments may correspond to the fibrillar network described in Xenopus oocyte nucleus being probably related to RNA transport. The molecular basis of this RNA remains elusive. Nevertheless, the morphological aspects of the spheroidal structures indicate they could correspond to nucleolar chromatin or to nucleolus-derived structures. We also speculate whether the complex aggregates of neighboring pores with intranuclear filaments may correspond to pore clustering previously described in these tetraploid animals using freeze-etching experiments. PMID- 17401477 TI - Elevated mazes as animal models of anxiety: effects of serotonergic agents. AB - This article reviews reported results about the effects of drugs that act upon the serotonergic neurotransmission measured in three elevated mazes that are animal models of anxiety. A bibliographic search has been performed in MEDLINE using different combinations of the key words X-maze, plus-maze, T-maze, serotonin and 5-HT, present in the title and/or the abstract, with no time limit. From the obtained abstracts, several publications were excluded on the basis of the following criteria: review articles that did not report original results, species other than the rat, intracerebral drug administration alone, genetically manipulated rats, and animals having any kind of experimental pathology. The reported results indicate that the effect of drugs on the inhibitory avoidance task performed in the elevated T-maze and on the spatio temporal indexes of anxiety measured in the X and plus mazes correlate with their effect in patients diagnosed with generalized anxiety disorder. In contrast, the drug effects on the one-way escape task in the elevated T-maze predict the drug response of panic disorder patients. Overall, the drug effects assessed with the avoidance task in the T-maze are more consistent than those measured through the anxiety indexes of the X and plus mazes. Therefore, the elevated T-maze is a promising animal model of generalized anxiety and panic disorder. PMID- 17401478 TI - Rhodnius prolixus Malpighian tubules and control of diuresis by neurohormones. AB - Rhodnius prolixus Malpighian tubules (MTs) are a good model for fluid and ion secretion studies in view of the dramatic postprandial diuresis, which follows its massive blood meals. Ingestion of a blood meal equals to 10-12 times their initial body mass, leads to rapid activation of high output by excretory system, which eliminates 40-50% of the fluid mass. Secretion of ions and water is stimulated 1000-fold by serotonin and diuretic hormone. These hormones cooperate synergistically to activate adenylate cyclase activity from MTs cells, which increase the level of intracellular cAMP. The anti-diuretic hormones have also an important role in the fluid maintenance of Rhodnius prolixus. Several hours after insect feeding occurs a reduction in urine flow, that has been thought to result from a decreased diuretic hormone release or from a novel mechanism of anti diuresis involving insect cardioacceleratory peptide 2b (CAP2b) and cyclic GMP. In this article it is discussed how the hormone regulation of fluid transport is done in Rhodnius prolixus MTs. PMID- 17401479 TI - Animal defense strategies and anxiety disorders. AB - Anxiety disorders are classified according to symptoms, time course and therapeutic response. Concurrently, the experimental analysis of defensive behavior has identified three strategies of defense that are shared by different animal species, triggered by situations of potential, distal and proximal predatory threat, respectively. The first one consists of cautious exploration of the environment for risk assessment. The associated emotion is supposed to be anxiety and its pathology, Generalized Anxiety Disorder. The second is manifested by oriented escape or by behavioral inhibition, being related to normal fear and to Specific Phobias, as disorders. The third consists of disorganized flight or complete immobility, associated to dread and Panic Disorder. Among conspecific interactions lies a forth defense strategy, submission, that has been related to normal social anxiety (shyness) and to Social Anxiety Disorder. In turn, Posttraumatic Stress Disorder and Obsessive-Compulsive Disorder do not seem to be directly related to innate defense reactions. Such evolutionary approach offers a reliable theoretical framework for the study of the biological determinants of anxiety disorders, and a sound basis for psychiatric classification. PMID- 17401480 TI - High prevalence of unusual genotypes of Toxoplasma gondii infection in pork meat samples from Erechim, Southern Brazil. AB - Toxoplasmosis is the most common cause of infectious uveitis in Brazil, with a higher frequency in the South of the country. We have collected samples from porcine tongue and diaphragm obtained in both large and small abattoirs and used molecular biological technique to determine the prevalence of infection and RFLP analysis to type the parasites. Seventeen out of 50 (34%) samples from the diaphragm and 33 out of 50 (66%) samples from the tongue demonstrated a positive PCR reaction for T. gondii and restriction analysis of four of the positive samples revealed that all had a type I genotype at SAG2. However, when other unlinked loci were analyzed, these strains had a type III genotype at markers BTUB, SAG3, and GRA6. One of the strains (8T) had a type II allele at SAG3, indicating it has a combination of alleles normally seen in the clonal lineages. Our sampling indicates a high prevalence of infection and suggests that unusual genotypes of T. gondii are found in Brazil even among domesticated pigs. PMID- 17401481 TI - Uranium-lead dating method at the Para-Iso isotope geology laboratory, UFPA, Belem - Brazil. AB - Analytical procedures for U-Pb isotope dilution analyses at the Para-Iso isotope geology laboratory of the Federal University of Para (UFPA) are described in detail. The procedures are applied to zircon, titanite, rutile, apatite, columbite-tantalite and whole rock. Reagent preparation and chemical processing are done in clean-room conditions. Samples are dissolved using Teflon microcapsules in steel jacket Teflon Parr Instrument bomb or Teflon screw cap containers. U and Pb are separated using anion exchange AG 1 x 8 resin columns. Typical blanks for mineral sample amounts of 0.01 to 1.0 mg are less than 1 pg U and 20-30 pg Pb. Isotope analysis of the U and Pb from the same filament are carried out using a Finnigan MAT 262 mass-spectrometer in static and dynamic modes. The current analytical level is demonstrated on analyses of international standard zircon 91500 with three different 235U-205Pb and 235U-208Pb isotope tracers and whole rock standards. Results of analyses of two zircon samples are also presented. PMID- 17401482 TI - Oceanographic parameters in continental margin of the State of Ceara (northeastern Brazil) deduced from C and O isotopes in foraminifers. AB - Specimens of Recent foraminifera of Amphistegina radiata, Peneroplis planatus and Globigerinoides ruber, from fifty samples of surface sediments of the continental margin of the State of Ceara, Brazil, have been analyzed for carbon and oxygen isotopes to investigate oceanographic parameters and determine the values of delta18O of the oceanic water. From a comparison between values of delta18O obtained for ocean water using the linear equations by (Craig and Gordon 1965) and the one by Wolff et al. (1998), it became evident that the former yielded a more reliable value (0.2 per thousand SMOW) than the latter. Lower values of delta18O for the ocean water in this continental margin resulted from continental water influence. Values of 18O (-0.3 per thousand to -1.5 per thousand PDB for benthic foraminifera and -0.6 per thousand to -2.4 per thousand PDB for planktic foraminifera), attest to a variation of temperatures of oceanic water masses, in average, between 20 to 22 degrees C in deep water and 24 to 27 degrees C, in surface water. Values of delta13C from +3.2% to -0.2 per thousand PDB (benthic foraminifera) reflect a variation in the apparent oxygen utilization (AOU) in the continental margin and indicate that the environments of bacteriological decomposition of organic matter are not continuous along the investigated area. PMID- 17401483 TI - Groundwater resources in the State of Sao Paulo (Brazil): the application of indicators. AB - Indicators, for groundwater resources, have mostly been employed to define the present status and the degradation tendency, regarding both quantity (under- or overexploitation) and quality (natural and anthropic contamination). This work presents the application of indicators in order to draw a picture of the groundwater resources situation in the 22 Water Resource Management Units (WRMU) of the State of Sao Paulo. The seven Indicators (I1 to I7) applied provide a general overview of groundwater dependence (I1, I2), availability (I3, I4), and quality (I5, I6, I7). Considering public supply (Indicator 1), one observes that 9 WRMUs show high (>50% of the population supplied by groundwater), 6, intermediate (49-25%), and 7, low (<24%) dependence on groundwater. Indicators 3 and 4 show that the resource still presents a great potential for further abstractions in most of the WRMUs, although there is evidence of overexploitation in the Upper Tiete, Turvo/Grande, and Pardo basins, and low availability in the Upper Tiete, Piracicaba/Capivari/Jundiai, and Turvo/Grande. Indicator 5 (aquifer natural vulnerability) denotes that the WRMUs 2, 4, 8, 13, 14 and 18-22 (part of the recharge area of Guarani Aquifer System) need more attention mainly where large contaminant loads are present. Indicator 6 shows the general excellent natural quality of groundwater, although it also denotes that 3 WRMUs need special consideration due to chromium and fluoride contamination. Indicator 7 demonstrates a close relationship between groundwater contamination occurrence and density/type of land occupation. PMID- 17401484 TI - The use of piezocone tests for high-resolution stratigraphy of Quaternary sediment sequences in the Brazilian coast. AB - Correlations between mapping units of costal sedimentary basin and interpretation of piezocone test results are presented and discussed based on examples from Caravelas strandplain, (State of Bahia), Paranagua (State of Parana) and Guaruja bays (State of Sao Paulo), Brazil. Recognizing that the sedimentary environment was mainly controlled by sea level fluctuations led to the interpretation of transgressive and regressive sedimentary sequences, which is in a good agreement with the sea level fluctuation curves currently accepted for these regions. The interpretation of piezocone test results shows that the sedimentary sequences of Caravelas and Guaruja sites are similar and they have a good correlation to the sea level fluctuation curve accepted for Salvador region, State of Bahia. On the other hand, the piezocone test results from Paranagua site indicate a different sedimentary sequence from the previous ones, relating to the sea level fluctuation curve accepted for Paranagua region. The results show the high applicability of piezocone testing for stratigraphical logging and suggest that it is possible to integrate it with other current techniques used for paleo environmental studies in Brazil, in accordance with recent approaches used in international research on the subject. PMID- 17401485 TI - Sedimentary changes on the Southeastern Brazilian upper slope during the last 35,000 years. AB - A first evaluation of the sedimentary changes, including accumulation of inorganic and organic carbon on the upper slope off Southeastern Brazil under different climatic conditions, based on the analysis of a sedimentary core, is here presented. Results indicate that sedimentation rate as well sedimentary characteristics, including the nature of the organic carbon deposited during the LGM (Last Glacial Maximum), were different from the present deposition, indicating higher primary productivity during the LGM, to be accounted for by the higher values of marine organic carbon. Nevertheless, the higher values of Accumulation Rate of calcium carbonate and organic carbon can be credited to the higher values of the sedimentation rates. Conditions prevailing during Isotope Stage 3 may be considered intermediate as between the LGM and the present day. The conditions for a higher primary productivity associated with the deposition of finer terrigenous sediments may be explained as a response to the off shoreward displacement of the main flow of the Brazil Current. PMID- 17401486 TI - Bone metabolism and vascular calcification. AB - Osteoporosis and atherosclerosis are chronic degenerative diseases which have been considered to be independent and whose common characteristic is increasing incidence with age. At present, growing evidence indicates the existence of a correlation between cardiovascular disease and osteoporosis, irrespective of age. The morbidity and mortality of osteoporosis is mainly related to the occurrence of fractures. Atherosclerosis shows a high rate of morbidity and especially mortality because of its clinical repercussions such as angina pectoris, acute myocardial infarction, stroke, and peripheral vascular insufficiency. Atherosclerotic disease is characterized by the accumulation of lipid material in the arterial wall resulting from autoimmune and inflammatory mechanisms. More than 90% of these fatty plaques undergo calcification. The correlation between osteoporosis and atherosclerosis is being established by studies of the underlying physiopathological mechanisms, which seem to coincide in many biochemical pathways, and of the risk factors for vascular disease, which have also been associated with a higher incidence of low-bone mineral density. In addition, there is evidence indicating an action of antiresorptive drugs on the reduction of cardiovascular risks and the effect of statins, antihypertensives and insulin on bone mass increase. The mechanism of arterial calcification resembles the process of osteogenesis, involving various cells, proteins and cytokines that lead to tissue mineralization. The authors review the factors responsible for atherosclerotic disease that correlate with low-bone mineral density. PMID- 17401487 TI - Description of Lyme disease-like syndrome in Brazil. Is it a new tick borne disease or Lyme disease variation? AB - An emerging clinical entity that reproduces clinical manifestations similar to those observed in Lyme disease (LD) has been recently under discussion in Brazil. Due to etiological and laboratory particularities it is named LD-like syndrome or LD imitator syndrome. The condition is considered to be a zoonosis transmitted by ticks of the genus Amblyomma, possibly caused by interaction of multiple fastidious microorganisms originating a protean clinical picture, including neurological, osteoarticular and erythema migrans-like lesions. When peripheral blood of patients with LD-like syndrome is viewed under a dark-field microscope, mobile uncultivable spirochete-like bacteria are observed. PCR carried out with specific or conservative primers to recognize Borrelia burgdorferi sensu stricto or the genus Borrelia has been negative in ticks and in biological samples. Two different procedures, respectively involving hematoxylin and eosin staining of cerebrospinal fluid and electron microscopy analysis of blood, have revealed spirochetes not belonging to the genera Borrelia, Leptospira or Treponema. Surprisingly, co-infection with microorganisms resembling Mycoplasma and Chlamydia was observed on one occasion by electron microscopy analysis. We discuss here the possible existence of a new tick-borne disease in Brazil imitating LD, except for a higher frequency of recurrence episodes observed along prolonged clinical follow-up. PMID- 17401488 TI - Exploratory calcineurin inhibitor-free regimens in living-related kidney transplant recipients. AB - Chronic allograft nephropathy is among the major causes of graft loss even in low risk kidney transplant recipients and correlates with acute nephrotoxic events during the first year post-transplant. Therefore, calcineurin inhibitor-free regimens may improve patient and graft survival among recipients of living related kidney transplants. To confirm this hypothesis, we evaluated the efficacy and safety of two calcineurin inhibitor-free regimens in 92 low-risk recipients of one-haplotype living-related kidney transplants. Immunosuppression consisted of tacrolimus, azathioprine and prednisone (group I, GI, N = 38), 2 doses of daclizumab, mycophenolate mofetil (MMF), and prednisone (GII, N = 33) and 2 doses of daclizumab, MMF, sirolimus and prednisone (GIII, N = 21). At 12 months, treatment failure (biopsy-confirmed acute rejection, graft loss or death) was higher in GII compared to GIII and GI (54.5 vs 24.0 vs 13.1%, P < 0.01, respectively). In patients of black ethnicity the incidence of acute rejection was 25 vs 83.3 vs 20% (P = 0.055), respectively. Patient and graft survival was comparable. There were no differences in mean creatinine or calculated creatinine clearance at 12 months. Overall incidence of post-transplant diabetes mellitus (3.3%) and cytomegalovirus disease (4.3%) was similar in all groups. Further development of effective calcineurin inhibitor-free regimens should exclude patients of black ethnicity and may need full-induction therapy, perhaps with depleting agents, and concentration-controlled use of sirolimus and MMF. PMID- 17401489 TI - Coronary fat content evaluated by morphometry in patients with severe atherosclerosis has no relation with serum lipid levels. AB - The relationship between lipid serum levels and coronary atherosclerotic plaque fat content was studied in 51 necropsy patients. Serum lipids were measured by standard techniques, during life, in the absence of lipid-lowering drugs. Intima, intimal fat and media areas were measured using a computerized system in cryosections of the odd segments of the right, anterior descending and circumflex coronary arteries stained with Sudan-IV. Mean intimal and lipid areas were 5.74 +/- 1.98 and 1.22 +/- 0.55 mm2 (22.12 +/- 8.48%) in 26 cases with high cholesterol (>or=200 mg/dL) and 4.98 +/- 1.94 and 1.16 +/- 0.66 mm2 (22.75 +/- 9.06%) in 25 cases with normal cholesterol (<200 mg/dL; P > 0.05). Patients with high levels of low-density lipoprotein (>or=130 mg/dL, N = 15) had a higher intima/media area ratio than those with normal levels of low-density lipoprotein (<130 mg/dL, N = 13, P < 0.01). No significant difference in the morphometrical variables was found in groups with high or low serum levels of triglycerides (>or=200 mg/dL, N = 13 vs <200 mg/dL, N = 36) or high-density lipoprotein (>or=35 mg/dL, N = 11 vs <35 mg/dL, N = 17). The association between the morphological measurements and serum levels of cholesterol, its fractions, and triglycerides was also tested and the correlation coefficients were low. Although high cholesterol is a risk factor, we show here that in patients with severe atherosclerosis blood cholesterol and triglyceride levels seem to have little influence on coronary lipid content, indicating that other factors may contribute to arterial lipid deposition and plaque formation. PMID- 17401490 TI - Association of the 894G>T polymorphism of the endothelial constitutive nitric oxide synthase gene with unstable angina. AB - The 894G>T polymorphism of the endothelial constitutive nitric oxide synthase gene consists of the substitution of a guanine base by a thymine at the 894th nucleotide of the gene. An association of this polymorphism with acute coronary syndromes has been described, only when in combination with other polymorphisms of this gene. The aim of the present study was to search for an association between this polymorphism and unstable angina in a southern Brazilian population. In a case-control study, 156 patients (group 1 (N = 83): unstable angina, group 2 (N = 73): stable angina) were genotyped by PCR and digestion of the product. Univariate analysis demonstrated that the minimal luminal diameter and the degree of stenosis of the culprit lesion differed between groups (P = 0.006 and 0.005, respectively). In addition, the frequencies of the T allele and of the T allele carriers (combined TT and TG genotypes) were significantly higher in the group with unstable angina (41.6 vs 28.8%; P = 0.025, Pearson chi-square test, and 73.5 vs 45.2%; P = 0.001, Pearson chi-square test, respectively). Multivariate logistic regression showed that the frequency of the T allele carriers was the only variable with a predictive value for unstable angina, when controlled for the other variables (6.1 (95% CI = 2.55-14.43); P < 0.001). Thus, in a homogenous group of patients, the endothelial constitutive nitric oxide synthase 894G>T polymorphism was associated with unstable angina. We suggest that this polymorphism may be a genetic risk factor for unstable angina. PMID- 17401491 TI - Correlations between pulse oximetry and peak expiratory flow in acute asthma. AB - Few studies are available concerning correlations between pulse oximetry and peak expiratory flow in children and adolescents with acute asthma. Although the Global Initiative for Asthma states that measurements of lung function and oximetry are critical for the assessment of patients, it is not clear if both methods should necessarily be included in their evaluation. Since there is a significant difference in cost between pulse oximetry equipment and peak expiratory flow devices, we determined whether clinical findings and peak expiratory flow measurements are sufficient to determine the severity of acute asthma. The present prospective observational study was carried out to determine if there is correlation between pulse oximetry and peak expiratory flow determination in 196 patients with acute asthma aged 4 to 15 years diagnosed according to the Global Initiative for Asthma criteria. Patients experiencing their first or second wheezing episode, with fever, related acute or chronic diseases, and unable to perform the peak expiratory flow maneuver were excluded. Measurements of peak expiratory flow and pulse oximetry were performed at admission and after 15 min of each inhaled salbutamol cycle. Correlations obtained by linear regression using the Pearson correlation coefficients (r) were 0.41 (P < 0.0001), 0.53 (P < 0.0001), 0.51 (P < 0.0001), and 0.61 (P < 0.0001) at admission and after the first, second and third cycles of salbutamol, respectively. These correlations showed that one measure cannot substitute the other (Pearson's coefficient <0.7), probably because they evaluate different aspects in the airways, suggesting that peak expiratory flow should not be used alone in the assessment of acute asthma in children and adolescents. PMID- 17401492 TI - Autonomic modulation of heart rate of young and postmenopausal women undergoing estrogen therapy. AB - The aim of the present study was to determine whether estrogen therapy (ET) reduces alterations of the autonomic control of heart rate (HR) due to hypoestrogenism and aging. Thirteen young (24 +/- 2.6 years), 10 postmenopausal (53 +/- 4.6 years) undergoing ET (PM-ET), and 14 postmenopausal (56 +/- 2.6 years) women not undergoing ET (PM) were studied. ET consisted of 0.625 mg/day conjugated equine estrogen. HR was recorded continuously for 8 min at rest in the supine and sitting positions. HR variability (HRV) was analyzed by time (SDNN and rMSSD indices) and frequency domain methods. Power spectral components are reported as normalized units (nu) at low (LF) and high (HF) frequencies, and as LF/HF ratio. Intergroup comparisons: SDNN index was higher in young (median: supine, 47 ms; sitting, 42 ms) than in PM-ET (33; 29 ms) and PM (31; 29 ms) women (P < 0.05). PM showed lower HFnu, higher LFnu and higher LF/HF ratio (supine: 44, 56, 1.29; sitting: 38, 62, 1.60) than the young group in the supine position (61, 39, 0.63) and the PM-ET group in the sitting position (57, 43, 0.75; P < 0.05). Intragroup comparisons: HR was lower in the supine than in the sitting position for all groups (P < 0.05). The HRV decrease from the supine to the sitting position was significant only in the young group. These results suggest that HRV decreases during aging. ET seems to attenuate this process, promoting a reduction in sympathetic activity on the heart and contributing to the cardioprotective effect of estrogen hormones. PMID- 17401493 TI - Comparison of anaerobic threshold determined by visual and mathematical methods in healthy women. AB - Several methods are used to estimate anaerobic threshold (AT) during exercise. The aim of the present study was to compare AT obtained by a graphic visual method for the estimate of ventilatory and metabolic variables (gold standard), to a bi-segmental linear regression mathematical model of Hinkley's algorithm applied to heart rate (HR) and carbon dioxide output (VCO2) data. Thirteen young (24 +/- 2.63 years old) and 16 postmenopausal (57 +/- 4.79 years old) healthy and sedentary women were submitted to a continuous ergospirometric incremental test on an electromagnetic braking cycloergometer with 10 to 20 W/min increases until physical exhaustion. The ventilatory variables were recorded breath-to-breath and HR was obtained beat-to-beat over real time. Data were analyzed by the nonparametric Friedman test and Spearman correlation test with the level of significance set at 5%. Power output (W), HR (bpm), oxygen uptake (VO2; mL kg(-1) min(-1)), VO2 (mL/min), VCO2 (mL/min), and minute ventilation (VE; L/min) data observed at the AT level were similar for both methods and groups studied (P > 0.05). The VO2 (mL kg(-1) min(-1)) data showed significant correlation (P < 0.05) between the gold standard method and the mathematical model when applied to HR (rs = 0.75) and VCO2 (rs = 0.78) data for the subjects as a whole (N = 29). The proposed mathematical method for the detection of changes in response patterns of VCO2 and HR was adequate and promising for AT detection in young and middle-aged women, representing a semi-automatic, non-invasive and objective AT measurement. PMID- 17401494 TI - Impact of marked weight loss induced by bariatric surgery on bone mineral density and remodeling. AB - Data about the impact of bariatric surgery (BS) and subsequent weight loss on bone are limited. The objective of the present study was to determine bone mineral density (BMD), bone remodeling metabolites and hormones that influence bone trophism in premenopausal women submitted to BS 9.8 months, on average, before the study (OGg, N = 16). The data were compared to those obtained for women of normal weight (CG, N = 11) and for obese women (OG, N = 12). Eight patients in each group were monitored for one year, with the determination of BMD, of serum calcium, phosphorus, magnesium, parathyroid hormone, 25 hydroxyvitamin D, insulin-like growth factor-I (IGF-I) and osteocalcin, and of urinary calcium and deoxypyridinoline. The biochemical determinations were repeated every three months in the longitudinal study and BMD was measured at the end of the study. Parathyroid hormone levels were similar in the three groups. IGF-I levels (CG = 332 +/- 62 vs OG = 230 +/- 37 vs OGg = 128 +/- 19 ng/mL) were significantly lower in the operated patients compared to the non-operated obese women. Only OGg patients presented a significant fall in BMD of 6.2% at L1-L4, of 10.2% in the femoral neck, and of 5.1% in the forearm. These results suggest that the weight loss induced by BS is associated with a significant loss of bone mass even at sites that are not influenced by weight overload, with hormonal factors such as IGF-I being associated with this process. PMID- 17401495 TI - Analysis of the diagnostic presentation profile, parathyroidectomy indication and bone mineral density follow-up of Brazilian patients with primary hyperparathyroidism. AB - Primary hyperparathyroidism is an endocrine disorder with variable clinical expression, frequently presenting as asymptomatic hypercalcemia in Western countries but still predominantly as a symptomatic disease in developing countries. The objective of this retrospective study was to describe the diagnostic presentation profile, parathyroidectomy indication and post-surgical bone mineral density follow-up of patients with primary hyperparathyroidism seen at a university hospital. We found 115 patients (92 women, median age 56 years) with primary hyperparathyroidism diagnosed during the last 20 years. We defined symptomatic patients based on the presence of any classical symptom affecting bone, kidney or the neuromuscular system. Surgical criteria followed the guidelines of the National Institutes of Health regarding asymptomatic primary hyperparathyroidism. Symptomatic patients and patients meeting surgical criteria for parathyroidectomy were 66 and 93% of the sample, respectively. Median calcium and parathyroid hormone values were 11.9 mg/dL and 189 pg/mL, respectively. After surgical treatment, 97% of patients were cured, with increases in bone mineral density of 19.4% in the lumbar spine and 15.7% in the femoral neck 3 years after surgery. Greater bone mass increases were detected in pre-menopausal women, men, and in symptomatic and younger patients, both in the lumbar spine and femoral neck. Our results support the previous findings of a predominantly symptomatic disease with a presentation profile that could be mainly related to a delayed diagnosis. Nevertheless, genetic and racial backgrounds, and nutritional factors such as calcium and vitamin D deficiency may play a role in the clinical presentation of primary hyperparathyroidism of Brazilian patients. PMID- 17401496 TI - Indoor NO2 air pollution and lung function of professional cooks. AB - Studies of cooking-generated NO2 effects are rare in occupational epidemiology. In the present study, we evaluated the lung function of professional cooks exposed to NO2 in hospital kitchens. We performed spirometry in 37 cooks working in four hospital kitchens and estimated the predicted FVC, FEV1 and FEF(25-75), based on age, sex, race, weight, and height, according to Knudson standards. NO2 measurements were obtained for 4 consecutive days during 4 different periods at 20-day intervals in each kitchen. Measurements were performed inside and outside the kitchens, simultaneously using Palm diffusion tubes. A time/exposure indicator was defined as representative of the cumulative exposure of each cook. No statistically significant effect of NO2 exposure on FVC was found. Each year of work as a cook corresponded to a decrease in predicted FEV1 of 2.5% (P = 0.046) for the group as a whole. When smoking status and asthma were included in the analysis the effect of time/exposure decreased about 10% and lost statistical significance. On predicted FEF(25-75), a decrease of 3.5% (P = 0.035) was observed for the same group and the inclusion of controllers for smoking status and asthma did not affect the effects of time/exposure on pulmonary function parameter. After a 10-year period of work as cooks the participants of the study may present decreases in both predicted FEV1 and FEF(25-75) that can reach 20 and 30%, respectively. The present study showed small but statistically significant adverse effects of gas stove exposure on the lung function of professional cooks. PMID- 17401497 TI - Comparison of the diagnosis of malaria by microscopy, immunochromatography and PCR in endemic areas of Venezuela. AB - Whole blood samples (N = 295) were obtained from different locations in Amazonas and Sucre States, in Venezuela. Malaria was diagnosed by microscopy, OptiMAL and polymerase chain reaction (PCR), with Plasmodium vivax, P. falciparum, and P. malariae being detected when possible. We identified 93 infections, 66 of which were caused by P. vivax, 26 by P. falciparum, and 1 was a mixed infection. No infection caused by P. malariae was detected. The sensitivity and specificity of each diagnostic method were high: 95.7 and 97.9% for microscopy, 87.0 and 97.9% for OptiMAL, and 98.0 and 100% for PCR, respectively. Most samples (72.2%) showed more than 5000 parasites/microL blood. The sensitivity of the diagnosis by microscopy and OptiMAL decreased with lower parasitemia. All samples showing disagreement among the methods were reevaluated, but the first result was used for the calculations. Parasites were detected in the 6 false-negative samples by microscopy after the second examination. The mixed infection was only detected by PCR, while the other methods diagnosed it as P. falciparum (microscopy) or P. vivax (OptiMAL) infection. Most of the false results obtained with the OptiMAL strip were related to the P. falciparum-specific band, including 3 species misdiagnoses, which could be related to the test itself or to genetic variation of the Venezuelan strains. The use of the microscopic method for malaria detection is recommended for its low cost but is very difficult to implement in large scale, population-based studies; thus, we report here more efficient methods suitable for this purpose. PMID- 17401498 TI - Genotyping hepatitis C virus from hemodialysis patients in Central Brazil by line probe assay and sequence analysis. AB - The present study examined the distribution of hepatitis C virus (HCV) genotypes and subtypes in a hemodialysis population in Goias State, Central Brazil, and evaluated the efficiency of two genotyping methods: line probe assay (LiPA) based on the 5' noncoding region and nucleotide sequencing of the nonstructural 5B (NS5B) region of the genome. A total of 1095 sera were tested for HCV RNA by RT nested PCR of the 5' noncoding region. The LiPA assay was able to genotype all 131 HCV RNA-positive samples. Genotypes 1 (92.4%) and 3 (7.6%) were found. Subtype 1a (65.7%) was the most prevalent, followed by subtypes 1b (26.7%) and 3a (7.6%). Direct nucleotide sequencing of 340 bp from the NS5B region was performed in 106 samples. The phylogenetic tree showed that 98 sequences (92.4%) were classified as genotype 1, subtypes 1a (72.6%) and 1b (19.8%), and 8 sequences (7.6%) as subtype 3a. The two genotyping methods gave concordant results within HCV genotypes and subtypes in 100 and 96.2% of cases, respectively. Only four samples presented discrepant results, with LiPA not distinguishing subtypes 1a and 1b. Therefore, HCV genotype 1 (subtype 1a) is predominant in hemodialysis patients in Central Brazil. By using sequence analysis of the NS5B region as a reference standard method for HCV genotyping, we found that LiPA was efficient at the genotype level, although some discrepant results were observed at the subtype level (sensitivity of 96.1% for subtype 1a and 95.2% for subtype 1b). Thus, analysis of the NS5B region permitted better discrimination between HCV subtypes, as required in epidemiological investigations. PMID- 17401499 TI - Effectiveness of highly active antiretroviral therapy using non-brand name drugs in Brazil. AB - In Brazil, HIV-infected individuals receive drugs (including non-brand name drugs which comprise locally produced generics and drugs that have not been tested in bioequivalence trials) free of charge from the government. The objective of the present study was to evaluate the effectiveness of highly active antiretroviral therapy (HAART) in Rio de Janeiro, Brazil, where non-brand drugs are widely used. For this purpose, we estimated the proportion of subjects with virologic failure (plasma HIV viral load greater than 400 copies/mL at 6 months after initiation of treatment). This was a retrospective cohort study of drug-naive HIV-infected subjects who initiated HAART. Subjects were included in the analysis if they were 18 years of age or older, were treatment naive, started HAART with a minimum of 3 drugs, and had available information on blood plasma HIV-1 viral load after 6 months on therapy. All subjects used antiretrovirals in dosing regimens recommended by the Brazilian National Advisory Committee for Antiretroviral Therapy. Chart reviews were conducted in three settings: at two public health outpatient units, at one clinical trial unit and at one private office. No comparisons of the effectiveness of non-brand name with the effectiveness of brand name drugs were made. We present results for 485 patients; of these, 354 (73%), 55 (11%), and 76 (16%) were seen at the public health outpatient units, private office, and clinical trial unit, respectively. Virologic failure was observed in 119 (25%) of the subjects. This study demonstrates the effectiveness of HAART in a setting where non-brand name drugs are widely used. PMID- 17401500 TI - Contribution of CD4+ T cells to the early mechanisms of ischemia- reperfusion injury in a mouse model of acute renal failure. AB - Renal ischemia-reperfusion (IR) injury is the major cause of acute renal failure in native and transplanted kidneys. Mononuclear leukocytes have been reported in renal tissue as part of the innate and adaptive responses triggered by IR. We investigated the participation of CD4+ T lymphocytes in the pathogenesis of renal IR injury. Male mice (C57BL/6, 8 to 12 weeks old) were submitted to 45 min of ischemia by renal pedicle clamping followed by reperfusion. We evaluated the role of CD4+ T cells using a monoclonal depleting antibody against CD4 (GK1.5, 50 micro, ip), and class II-major histocompatibility complex molecule knockout mice. Both CD4-depleted groups showed a marked improvement in renal function compared to the ischemic group, despite the fact that GK1.5 mAb treatment promoted a profound CD4 depletion (to less than 5% compared to normal controls) only within the first 24 h after IR. CD4-depleted groups presented a significant improvement in 5-day survival (84 vs 80 vs 39%; antibody treated, knockout mice and non depleted groups, respectively) and also a significant reduction in the tubular necrosis area with an early tubular regeneration pattern. The peak of CD4 positive cell infiltration occurred on day 2, coinciding with the high expression of betaC mRNA and increased urea levels. CD4 depletion did not alter the CD11b infiltrate or the IFN-gamma and granzyme-B mRNA expression in renal tissue. These data indicate that a CD4+ subset of T lymphocytes may be implicated as key mediators of very early inflammatory responses after renal IR injury and that targeting CD4+ T lymphocytes may yield novel therapies. PMID- 17401501 TI - TNF-alpha and IL-10 levels in tracheobronchial lavage of ventilated preterm infants and subsequent lung function. AB - The role of airway inflammation in ventilated preterm newborns and the risk factors associated with the development of chronic lung disease are not well understood. Our objective was to analyze the association of the airway inflammatory response in ventilated preterm infants by serial measurements of TNF alpha and IL-10 in tracheobronchial lavage (TBL) with perinatal factors and lung function measured early in life. A series of TBL samples were collected from ventilated preterm infants (less than 32 weeks of gestational age) and concentrations of TNF-alpha and IL-10 were measured by ELISA. Pulmonary function tests were performed after discharge by the raised volume rapid compression technique. Twenty-five subjects were recruited and 70 TBL samples were obtained. There was a significant positive association between TNF-alpha and IL-10 levels and length of time between the rupture of the amniotic membranes and delivery (r = 0.65, P = 0.002, and r = 0.57, P < 0.001, respectively). Lung function was measured between 1 and 22 weeks of corrected age in 10 patients. Multivariable analysis with adjustment for differences in lung volume showed a significant negative association between TNF-alpha levels and forced expiratory flow (FEF(50); r = -0.6; P = 0.04), FEF(75) (r = -0.76; P = 0.02), FEF(85) (r = -0.75; P = 0.03), FEF(25-75) (-0.71; P = 0.02), and FEV(0.5) (r = -0.39; P = 0.03). These data suggest that TNF-alpha levels in the airways during the first days of life were associated with subsequent lung function abnormalities measured weeks or months later. PMID- 17401502 TI - Cortisol and 17-alpha-hydroxy-progesterone levels in infants with refractory hypotension born at 30 weeks of gestation or less. AB - Refractory hypotension is frequent in very low-birth weight infants, whose hypothalamic-pituitary-adrenal axis has been suggested to be immature. The objective of the present study was to evaluate basal cortisol and 17-alpha-OH progesterone in the first 36 h of life in preterm infants with and without refractory hypotension (mean arterial blood pressure below the lower limit for gestational age throughout the study despite aggressive volume expansion and use of vasopressors). Thirty-five infants with 5 mm) and MBs on baseline and follow-up GRE were evaluated. The relationship of recurrent ICHs with initial and follow-up clinical and laboratory data as well as the MBs was assessed. RESULTS: Among 63 patients, 43 (68.3%) had MBs (median, 2; range, 1 to 17) on baseline GRE. Seven (11.1%) patients (6 with initial MBs; 1 without initial MBs) developed recurrent ICHs, and 19 (30.2%) had new MBs during a median 23.3 months (range, 8.3 to 33.0) of follow-up. The number of initial MBs on baseline GRE was significantly (p < 0.0001) associated with development of recurrent ICHs whereas other clinical and laboratory data were not. CONCLUSIONS: Recurrent ICHs and MBs are common after long-term follow-up of primary ICH. The number of MBs on baseline GRE may predict the recurrence of the ICH. PMID- 17401518 TI - Transcranial color-coded sonography helps differentiation between idiopathic Parkinson's disease and vascular parkinsonism. AB - BACKGROUND: Recently, transcranial color-coded sonography (TCCS) has been found to have a diagnostic value in patients with idiopathic Parkinson's disease (IPD), which displays increased hyperechogenicity at the substantia nigra (SN). OBJECTIVE: To use TCCS, to assess the difference in SN hyperechogenicity and intracranial hemodynamics among subjects with IPD, vascular parkinsonism (VP) and controls. METHODS: Eighty IPD and 30 VP patients, and 60 controls were recruited into this study. The hyperechogenicity area at the SN and midbrain were calculated by encircling the outer circumference from the ipsilateral temporal window, using TCCS in each subject. The hemodynamics of intracranial large arteries, including flow velocity and pulsatility index (PI), were also measured. RESULTS: The presence of SN hyperechogenicity was significantly higher in the IPD patients than in the VP patients and controls (84% vs. 20% & 5%, respectively, p < 0.001). In IPD patients, the SN hyperechogenicity was correlated with the neurological severity and disease duration. Twenty-five (66.7%) VP patients had obvious vascular abnormality, as seen in TCCS study. The mean PI was significantly more elevated in the VP patients than those in the IPD patients and controls (all p < 0.05), but there was no significant difference of flow velocities among the VP, IPD patients and controls. CONCLUSION: TCCS, combining B mode imaging for SN echogenicity and trancranial Doppler for intracranial hemodynamics, is a useful diagnostic tool in the differentiation between IPD and VP. These findings also suggest that multiple subcortical vascular lesions may damage the basal ganglia and thalamocortical circuit and result in parkinsonism features in VP patients. PMID- 17401519 TI - Varicella-zoster virus at relapses of multiple sclerosis. AB - The possible participation of different herpes viruses was studied during exacerbations of multiple sclerosis (MS). We searched for the presence of DNA from the following herpes viruses: varicella zoster virus (VZV), herpes-simplex viruses 1 and 2; Epstein-Barr virus (EBV) and human herpes-virus-6 (HHV6) in mononuclear cells from patients with MS during relapse (n = 40), MS during remission (n = 131) and controls (n = 125). Additionally, immune cells containing viral antigens were quantified by flow cytometry, and VZV load was determined by real time PCR in 2 MS patients at various times during relapse and remission. DNA from VZV was found in 95% of MS patients during relapse and in 17% during remission; all controls were negative; by contrast, DNA from HHV6 was found in 24% of MS patients during relapse and in 2% during remission; DNA from herpes simplex viruses was not found in any subject; and DNA from EBV was found in a similar percentage of subjects from all groups. Sequential quantification of VZV load showed a curve that increased during relapse and disappeared at remission. Also, VZV antigens were found inside a large number of immune cells from MS patients during relapse as compared with MS patients on remission and controls. In the typical forms of VZV infection, varicella and herpes-zoster, DNA from VZV is found in mononuclear cells exclusively during brief periods at the beginning of the active infection, but not during latency; thus, the conspicuous presence of VZV during relapses of MS may indicate a period of active infection and suggests the participation of VZV in the pathogenesis of MS. PMID- 17401520 TI - Two subtypes of optic-spinal form of multiple sclerosis in Japan: clinical and laboratory features. AB - Seventy-seven cases of the optic-spinal form of multiple sclerosis (OSMS) were collected from 6 institutes in 3 cities of Japan, and the clinical and MRI features were analyzed. Two-thirds of the OSMS patients had longitudinally extensive spinal cord MRI lesions (LESL), and had clinical features similar to those of relapsing neuromyelitis optica which often causes severe disability. In contrast, OSMS patients without LESL tended to have milder disease and had some feature commonly seen in the conventional form of MS. The percentage of OSMS without LESL in total OSMS has recently been increasing. The present study suggests that LESL is crucially important for distinguishing the two subtypes of OSMS. PMID- 17401521 TI - Investigation of the mitochondrial genome in patients with atypical motor neuron disease. AB - The molecular aetiology of many patients with motor neuron disease (MND) remains unknown. Recent evidence of mitochondrial dysfunction, in particular the finding of histochemical abnormalities and pathogenic mitochondrial DNA (mtDNA) mutations, has prompted us to investigate further the role of mtDNA abnormalities in a cohort of thirteen patients with atypical MND presentations by whole mitochondrial genome sequencing. No pathogenic mutations were detected suggesting that inherited mtDNA mutations are not a common cause of atypical MND presentations. PMID- 17401522 TI - MRI derived brain atrophy in PSP and MSA-P. Determining sample size to detect treatment effects. AB - Progressive supranuclear palsy (PSP) and multiple system (MSA) atrophy are associated with progressive brain atrophy. Serial MRI can be applied in order to measure this change in brain volume and to calculate atrophy rates. We evaluated MRI derived whole brain and regional atrophy rates as potential markers of progression in PSP and the Parkinsonian variant of multiple system atrophy (MSA P). 17 patients with PSP, 9 with MSA-P and 18 healthy controls underwent two MRI brain scans. MRI scans were registered, and brain and regional atrophy rates (midbrain, pons, cerebellum, third and lateral ventricles) measured. Sample sizes required to detect the effect of a proposed disease-modifying treatment were estimated. The effect of scan interval on the variance of the atrophy rates and sample size was assessed. Based on the calculated yearly rates of atrophy, for a drug effect equivalent to a 30% reduction in atrophy, fewer PSP subjects are required in each treatment arm when using midbrain rather than whole brain atrophy rates (183 cf. 499). Fewer MSA-P subjects are required, using pontine/cerebellar, rather than whole brain atrophy rates (164/129 cf. 794). A reduction in the variance of measured atrophy rates was observed with a longer scan interval. Regional rather than whole brain atrophy rates calculated from volumetric serial MRI brain scans in PSP and MSA-P provide a more practical and powerful means of monitoring disease progression in clinical trials. PMID- 17401523 TI - Functional outcome after severe cerebral venous thrombosis. AB - Severe cerebral venous thrombosis (CVT) is a rare cerebrovascular condition which in the more severe cases warrants intensive care treatment. While the outcome in the majority of uncomplicated CVT cases is good, it may be fatal in more affected patients. We provide long-term functional and quality of life (QOL) outcome data in the form of a retrospective analysis of 10 patients admitted to a neurological ICU with severe CVT. Outcome measures used were the modified Rankin Scale, the 36 item Short Form Health Survey, and the Psychological General Well-Being index. The mortality rate was 50% but 4 out 5 survivors had a good functional outcome with normal QOL despite a very severe clinical course. This finding justifies extensive life-sustaining therapy as the prognosis even of severe cases may be good if the acute phase is survived. PMID- 17401524 TI - A population based study of respiratory function in motor neuron disease patients living in Tayside and North East Fife, Scotland. AB - Respiratory failure is a major cause of morbidity and the principal cause of death in motor neuron disease; non-invasive ventilation is increasingly used worldwide to palliate the respiratory symptoms. This observational study was designed to evaluate the prevalence of respiratory insufficiency within the motor neuron disease population of Tayside and North East Fife, Scotland. Twenty-six patients were identified, their diagnosis confirmed according to agreed criteria and subjected to the Revised Amyotrophic Lateral Sclerosis Functional Rating Scale, the Epworth Sleepiness questionnaire; spirometry, sniff nasal inspiratory pressure and nocturnal pulse oximetry measurements.Twenty-two (84.6%) patients reported one or more symptoms of respiratory insufficiency, 19 patients (73%) had forced vital capacity <80% of predicted in the sitting position and 10 (38.5%) had oxygen saturation <90% for >5% of night. On this basis a potential 10 patients required consideration for ventilation. As well as probable improvement in quality of life and survival for those patients this potential increase in workload has major educational, management and resource implications for health care providers. PMID- 17401525 TI - Relationship between age and subtypes of psychotic symptoms in Parkinson's disease. AB - OBJECTIVE: Psychotic symptoms (PS) in Parkinson's disease (PD) usually develop as a side effect of the dopaminergic therapy and consist of hallucinations and delusions. We observed that PD patients who developed delusions tend to be younger than those with hallucinations and we aimed to investigate the validity of this observation. METHODS: The medical records of 127 PD patients with PS were reviewed and 76 patients who were on treatment with dopamine agonists with or without levodopa at the time of developing PS were included. Patients were stratified into 3 groups according to the subtypes of PS: patients with solely hallucinations (n = 46), solely delusions (n = 18), and both types (n = 12). The groups were compared with respect to the age-at-onset of PD and PS, duration of PD, Activities of Daily Living (ADL) and motor subscale scores of Unified PD Rating Scale (UPDRS), and levodopa equivalent dose of the dopaminergic agents administered at the time of PS onset. RESULTS: The mean age-at onset of PD and PS was significantly younger (p = 0.0001) in patients with delusions (49 and 55.9 years) than those with hallucinations (61.9 and 68.9 years). The same parameters were also significantly different (p = 0.002 and p = 0.001, respectively) between the groups of patients with concurrent delusions and hallucinations (51.7 and 57.2 years) and those with only hallucinations. ADL and motor subscale scores were higher in patients with hallucinations (p = 0.016 and p = 0.013) compared with those noted in patients with delusions despite similar disease duration. The mean levodopa equivalent doses of the dopaminergic agents administered at the time of onset of PS did not differ between the groups. CONCLUSION: This study supported an association of delusions with younger onset of both PD and psychosis as compared with hallucinations. However, additional factors related to this association remain to be elucidated. PMID- 17401526 TI - Late onset vanishing white matter disease. PMID- 17401527 TI - Bilateral language function in callosal agenesis: an fMRI and DTI study. PMID- 17401528 TI - Interaction between 5-HT1A and BDNF genotypes increases the risk of treatment resistant depression. AB - Several studies have linked 5-HT1A C1019G and BDNF G196A (Val66Met) gene polymorphisms to major depressive disorder (MDD) and the actions of antidepressants. We attempt to show that the interaction between 5-HT1A and BDNF polymorphism predicts the risk of treatment-resistant depression. The sample consists of 119 patients with treatment-resistant MDD and 392 controls. 5-HT1A C1019G and BDNF G196A (Val66Met) polymorphisms were studied. The combination of 5 HT1A GG and BDNF GA + AA genotypes is associated with an increased risk of depression. PMID- 17401529 TI - Effect of aldehydes derived from oxidative deamination and oxidative stress on beta-amyloid aggregation; pathological implications to Alzheimer's disease. AB - Formaldehyde and methylglyoxal are generated via deamination from methylamine and aminoacetone respectively catalyzed by semicarbazide-sensitive amine oxidase (SSAO). Malondialdehyde (MDA) and 4-hydroxynonenal (HNE) are end products of lipid peroxidation due to oxidative stress. These aldehydes are capable of inducing protein cross-linkage. Elevated levels of aldehydes were found in Alzheimer's disease (AD). These reactive metabolites may potentially play important roles in beta-amyloid (Abeta) aggregation related to the pathology of AD. In the present study thioflavin-T (ThT) fluorometry, an immuno-dot-blot assay and atomic force microscopy (AFM) were employed to reveal the effect of aldehydes on Abeta aggregation in vitro. The target on Abeta for interaction with formaldehyde was identified. The results support the involvement of endogenous aldehydes in amyloid deposition related to AD. PMID- 17401530 TI - Factors affecting the relative importance of amine oxidases and monooxygenases in the in vivo metabolism of xenobiotic amines in humans. AB - The monooxygenases and the amine oxidases (AOs) are the major enzyme systems involved in vivo in the oxidative metabolism of xenobiotic amines in humans. With the exception of the inhibition of the metabolism of tyramine ingested by subjects taking inhibitors of MAO-A or of both MAO-A and -B, which has been extensively investigated, the involvement of the monoamine oxidases in xenobiotic amine metabolism (drugs in particular) has been largely neglected. Furthermore, with the exception of amlodipine, there have been essentially no studies on the metabolism of drug amines by amine oxidases such as SSAOs and PAOs in humans. In contrast, monooxygenases (CYP isoenzymes, and to a lesser extent, FMOs) have been extensively investigated in terms of their involvement in xenobiotic metabolism. It is possible that the contribution of AOs to the overall metabolism of xenobiotic amines in humans has been underestimated, or erroneously estimated, as most investigations of drug metabolism have been performed using in vitro test systems optimized for CYP activity, such as liver microsomes, and most investigations of drug metabolism in vivo in humans have identified only the final, stable metabolites. PMID- 17401531 TI - Hydrogen peroxide derived from amine oxidation mediates the interaction between aminosugars and semicarbazide-sensitive amine oxidase. AB - Semicarbazide-sensitive amine oxidase (SSAO) also functions as a vascular adhesion protein (VAP-1). The nature of the target site on lymphocytes to which endothelial-cell SSAO/VAP-1 binds is unknown. We have shown that amino sugars (galactosamine, glucosamine and mannosamine), which are not SSAO substrates, can bind to the enzyme as reversible inhibitors. Thus, they serve as a model system in which to study the interaction process. Binding occurred during substrate (benzylamine) oxidation but not when the amino sugar was incubated, for extended periods, with SSAO alone. These results suggest that one, or more of the products of the SSAO-catalysed amine oxidation might be necessary for the inhibitory process to occur. Two of the reaction products of benzylamine oxidation, benzaldehyde and ammonia were found to have no effect on the inhibition of SSAO by galactosamine. Preincubation of the enzyme with galactosamine plus H(2)O(2) was, however, found to result in time-dependent inhibition. This is not a result of the non-enzymic reaction between H(2)O(2) and the amino sugar, since preincubation of galactosamine with H(2)O(2) alone, for extended periods, did not give rise to an inhibitory species. The amount of exogenously added H(2)O(2) necessary for inhibition was very much greater than that formed during substrate oxidation. These results suggest that the H(2)O(2) formed as a product of the SSAO-catalysed oxidation reaction is more efective in promoting the binding of amino sugars. PMID- 17401532 TI - The effects of buffer cations on interactions between mammalian copper-containing amine oxidases and their substrates. AB - We and others have observed that substrates for copper-containing amine oxidases cause substrate inhibition at high concentrations. Through use of a novel "pseudoquantitative" rapid equilibrium approach, kinetic analyses with human and bovine enzymes indicate that these effects are consistent with substrates binding to oxidised and reduced enzyme forms. Small cations compete with binding of substrates to oxidised and reduced enzyme, influencing both substrate turnover and substrate inhibition patterns. Cations reduce affinity of the resting bovine enzyme for spermidine, but not benzylamine, indicating that the predominant effect of cations on substrate oxidation results from binding to an anionic site outside the active site. However, binding of cations to the active site of the reduced form of both enzymes attenuates substrate inhibition with both spermidine and benzylamine. Our observations have significant practical implications for researchers assaying kinetic behaviour of these enzymes, and particularly those developing novel inhibitors of human copper-containing amine oxidases. PMID- 17401533 TI - Docking of novel reversible monoamine oxidase-B inhibitors: efficient prediction of ligand binding sites and estimation of inhibitors thermodynamic properties. AB - Monoamine oxidase (MAO, EC 1.4.3.4) is a flavoenzyme bound to the mitochondrial outer membranes of the cells, which is responsible for the oxidative deamination of neurotransmitter and dietary amines. It has two distinct isozymic forms, designated MAO-A and MAO-B, each displaying different substrate and inhibitor specificities. They are the well-known target for antidepressant, Parkinson's disease and neuroprotective drugs. Elucidation of the x-ray crystallographic structure of MAO-B has opened the way for molecular modeling studies. In this research 12 reversible and MAO-B selective inhibitors have been docked computationally to the active site of the MAO-B enzyme. AutoDock 3.0.5 was employed to perform the automated molecular docking. The result of docking studies generated thermodynamic properties, such as free energy of bindings (DeltaG(b)) and inhibition constants (K (i)) for the inhibitors. Moreover, 3D pictures of inhibitor-enzyme complexes afforded valuable data regarding the binding orientation of each inhibitor in the active site of MAO-B. PMID- 17401534 TI - Do monomeric vs dimeric forms of MAO-A make a difference? A direct comparison of the catalytic properties of rat and human MAO-A's. AB - The recent crystallographic structures of human MAO-A and rat MAO-A have shown that human MAO-A is monomeric whereas rat MAO-A is a dimer, even though they share approximately 90% sequence identity. The functional significance of this structural difference is unknown. Therefore, biochemical approaches in this paper were performed to investigate the influence of oligomeric state on functional properties of human and rat MAO-A's. The data show that 1) dimerization of MAO-A increases its structural stability; 2) the differences in kinetic properties may be caused by differences in active site structures as a result of differences in oligomeric states of the human and the rat enzymes; 3) QSAR studies show rat MAO A as well as human MAO-A catalysis occur via proton abstraction mechanisms, and the binding of substrates is similar for both enzymes. PMID- 17401535 TI - Variations in activity and inhibition with pH: the protonated amine is the substrate for monoamine oxidase, but uncharged inhibitors bind better. AB - It has been accepted that, as required mechanistically, the neutral form of the amine is the substrate for monoamine oxidase, despite the amine pK (a) of above 9.5. The pH dependence of the kinetic parameters for kynuramine oxidation by purified human MAO-A and for phenylethylamine oxidation by MAO-B in granulocytes at pH values from 5 to 10 was consistent with the protonated amine being used. Deprotonation of a group of pK (a) = 7.1 in MAO-B and pK (a) = 7.5 +/- 0.1 (n = 4) in MAO-A was important for efficient catalysis. The K(i) values for two oxazolidinone inhibitors of MAO-A gave opposite pH-dependence indicating that the uncharged form of each inhibitor bound better than the charged form. Decreased pH induced a blue shift in the spectral maximum of MAO-A indicative of a more hydrophobic environment around the flavin, and also influenced the redox properties of the flavin. PMID- 17401536 TI - The aromatic cage in the active site of monoamine oxidase B: effect on the structural and electronic properties of bound benzylamine and p-nitrobenzylamine. AB - Computational studies using the ONIOM methods have been performed to probe the catalytic roles of tyrosine residues 398 and 435 which constitute the "aromatic cage" in the active site of MAO-B. The results presented here provide additional new insights into the interactions that take place on activation of the amine substrate by the aromatic cage residues in MAO-B catalysis and have relevance to the MAO-A catalytic mechanism. PMID- 17401537 TI - Short and long term changes in NMDA receptor binding in mouse brain following chronic phencyclidine treatment. AB - Phencyclidine (PCP) is an antagonist of the NMDA subtype of glutamate receptor. PCP treatment induces psychosis in normal humans, which provides a valuable model of schizophrenia. PCP administration also models some of the symptoms of schizophrenia in experimental animals. NMDA hypofunction has been hypothesized to explain these schizophrenia-like symptoms. Acute or chronic administration of the NMDA receptor antagonist PCP has been shown to induce several short or long-term effects in both humans and experimental animals. In an attempt to clarify the neurochemical substrates of these effects in the present study, we used quantitative autoradiography to examine the effects of chronic (14 days) PCP treatment on NMDA receptor binding in mouse brain following both a short- (1 and 24 h) and long-term (14 days) delay after the last PCP treatment. NMDA receptors were targeted using [(3)H]MK801. Chronic PCP treatment increased [(3)H]MK801 binding consistently in the hippocampus in the short-term (p < 0.01). Conversely in the long-term, there were widespread reductions in NMDA receptor binding and this effect was most evident in the hippocampus where a 35% reduction of binding was found (p < 0.001). These results suggest that the hippocampus has a strong involvement in both the short and long-term effects of PCP treatment and that reduced NMDA receptor function might be one of the neurochemical substrates of the long lasting actions of PCP or PCP-induced psychosis. Importantly, this study shows that the long-term delay following chronic PCP treatment more accurately represents a state of NMDA hypofunction than the short-term PCP model. PMID- 17401538 TI - Chronic dose effects of reboxetine on motor skill acquisition and cortical excitability. AB - BACKGROUND: Enhancement of cortical excitability is thought to be beneficial for synaptic plasticity associated with motor skill acquisition. Single dose application of the selective norepinephrine reuptake inhibitor reboxetine (RBX) increases motor cortex excitability. In this study, we tested if a chronic dose application of RBX improved motor skill acquisition and modulated cortical excitability. METHODS: The study was randomised, double blind and placebo controlled. Twelve healthy subjects received four milligram RBX twice a day for four days preceded by two milligram RBX twice a day for two days. Each subject served as his own control. The time interval between the verum and the placebo session was 16 days or more. Measurement of cortical excitability by means of paired pulse transcranial magnetic stimulation (ppTMS) was conducted before and after the motor skill acquisition task in each session. The task was to lift two fingers of the right hand at once while the hand was positioned sprawled out on the table. The movements were self-paced and subjects had to perform as many moves as possible in 60 sec. Between seven blocks of self-paced movements six blocks with 60 single trials at a fixed interstimulus intervall were presented. Two equally difficult versions of the task using different finger combinations were established in order to avoid carry over effects in performance. The finger movements were recorded with a three-dimensional ultrasound movement analysis system (Zebris). RESULTS: All subjects had substantial gain in performance across the selfpaced blocks. Average increase in number of correct moves was 87% (from 27.8 to 51.9). There was no significant difference neither between the versions of the task nor between placebo vs. verum. Also, there was no significant difference between first and second session, indicating that there was no carry over effect in performance. ppTMS revealed no significant differences in cortical excitability between groups. CONCLUSION: The newly developed skill acquisition task yields robust single subject gain of performance. As the two versions of the task do not interact, it is suitable to be used in cross-over designs. In contrast to studies using single doses of RBX, motor cortex excitability seems to be unaffected in a steady-state induced by repeated drug applications. This could explain why RBX did not modulate motor behavior. PMID- 17401539 TI - Plasma amino acids and neopterin in healthy persons with Down's syndrome. AB - In persons with Down's syndrome (DS) immunological abnormalities as well as hypothyroidism and Alzheimer type dementia are frequently observed. In addition, the activity of the enzyme cystathionine beta-synthase (CBS) is over-expressed which results in an altered homocysteine metabolism. In the present study, 48 older healthy DS persons without signs of dementia, psychiatric or somatic comorbidity and free of medication were analyzed for plasma levels of amino acids, neopterin and monoaminergic metabolites. Data were compared with those obtained from age and sex matched healthy controls. It was found that the spectrum of amino acids showed widespread differences in that levels of nearly all essential amino acids were lower in DS patients as compared to healthy controls. In addition, a significantly lower methionine and higher taurine concentration were observed which is in accordance with a disturbed homocysteine metabolism. With respect to the monoamine metabolites, the concentration of 5 hydroxyindoleacetic acid was not altered whereas that of homovanillic acid was significantly increased. Finally, the concentration of the immune activation marker neopterin was increased in persons with DS. It is concluded that healthy DS persons of older age show extensive biochemical abnormalities suggesting a compromised homocysteine metabolism, an activated cell-mediated immune response and an enhanced turnover of dopamine. PMID- 17401540 TI - Phylotype diversity of deep-sea hydrothermal vent prokaryotes trapped by 0.2- and 0.1-microm-pore-size filters. AB - Eleven 16S rRNA gene clone libraries including 34 archaeal and 72 bacterial phylotypes were constructed from total 708 clones of hydrothermal vent prokaryotes trapped by 0.2- and 0.1-microm-pore-size filters. Crenarchaeota and Proteobacteria phylotypes dominated the archaeal and bacterial populations, respectively. Novel unaffiliated phylotypes occurred only in the 0.1-microm trapped populations. PMID- 17401541 TI - The isolation and initial characterization of mercury resistant chemolithotrophic thermophilic bacteria from mercury rich geothermal springs. AB - Mercury rich geothermal springs are likely environments where mercury resistance is critical to microbial life and where microbe-mercury interactions may have evolved. Eleven facultative thermophilic and chemolithoautotrophic, thiosulfate oxidizing bacteria were isolated from thiosulfate enrichments of biofilms from mercury rich hot sulfidic springs in Mount Amiata, Italy. Some strains were highly resistant to mercury (>or=200 muM HgCl(2)) regardless of its presence or absence during primary enrichments, and three reduced ionic mercury to its elemental form. The gene encoding for the mercuric reductase enzyme (MerA), was amplified by PCR from seven strains. However, one highly resistant strain did not reduce mercury nor carried merA, suggesting an alternative resistance mechanism. All strains were members of the order Bacillales and were most closely related to previously described thermophiles belonging to the Firmicutes. Phylogenetic analyses clustered the MerA of the isolates in two supported novel nodes within the Firmicutes lineage and a comparison with the 16S rRNA gene tree suggested at least one case of horizontal gene transfer. Overall, the results show that the thermophilic thiosulfate oxidizing isolates were adapted to life in presence of mercury mostly, but not exclusively, by possessing MerA. These findings suggest that reduction of mercury by chemolithotrophic thermophilic bacteria may mobilize mercury from sulfur and iron deposits in geothermal environments. PMID- 17401542 TI - Thermal stability of isocitrate dehydrogenase from Archaeoglobus fulgidus studied by crystal structure analysis and engineering of chimers. AB - Isocitrate dehydrogenase from Archaeoglobus fulgidus (AfIDH) has an apparent melting temperature (T(m)) of 98.5 degrees C. To identify the structural features involved in thermal stabilization of AfIDH, the structure was solved to 2.5 A resolution. AfIDH was strikingly similar to mesophilic IDH from Escherichia coli (EcIDH) and displayed almost the same number of ion pairs and ionic networks. However, two unique inter-domain networks were present in AfIDH; one three membered ionic network between the large and the small domain and one four membered ionic network between the clasp and the small domain. The latter ionic network was presumably reduced in size when the clasp domain of AfIDH was swapped with that of EcIDH and the T (m) decreased by 18 degrees C. Contrarily, EcIDH was only stabilized by 4 degrees C by the clasp domain of AfIDH, a result probably due to the introduction of a unique inter-subunit aromatic cluster in AfIDH that may strengthen the dimeric interface in this enzyme. A unique aromatic cluster was identified close to the N-terminus of AfIDH that could provide additional stabilization of this region. Common and unique heat adaptive traits of AfIDH with those recently observed for hyperthermophilic IDH from Aeropyrum pernix (ApIDH) and Thermotoga maritima (TmIDH) are discussed herein. PMID- 17401543 TI - A simple method to determine concentration of enantiomers in enzyme-catalyzed kinetic resolution. AB - Kinetic resolutions play important roles in industrial biotransformations for production of optical pure compounds from racemic substrates. A simple method, based on enantiomeric excess of both substrate (ee (S)) and the corresponding product (ee (P)), was developed for determination of concentration of enantiomers in kinetic resolution. Since only relative quantity (ee) was required in the proposed method, calibration and cumbersome quantitative sample handling can be avoided and analytical accuracy can be greatly improved. PMID- 17401544 TI - Purification and characterization of prokaryotically expressed human interferon lambda2. AB - A system for the production of soluble interferon (IFN)-lambda2 was developed by fusing the IFN-lambda2, NusA protein, polyhistidine and S peptide genes and then expressing the fused product (Nus-His-S-tagged IFN-lambda2) in Escherichia coli. The expressed fusion protein was purified by Ni-NTA affinity chromatography. The fusion tag was removed from recombinant IFN-lambda2 by cleavage with enterokinase. N-Terminal sequencing confirmed the identity of the purified protein. When compared with commercial IFN-alpha2b, IFN-lambda2 had similar antiviral activity. The production and characterization of biologically active IFN-lambda2 will be beneficial for its potential role in clinical applications. PMID- 17401545 TI - Secretory fluorescent protein, a secretion green fluorescent fusion protein with alkaline phosphatase activity as a sensitive and traceable reporter in baculovirus expression system. AB - The advantages of using traceable fluorescent protein (enhanced green fluorescent protein; EGFP) and a secretory alkaline phosphatase (SEAP) have been used to generate a reporter gene: the secretory fluorescent protein (SEFP). Sf21 cells, infected with the recombinant baculovirus containing the SEFP gene, revealed both traceable fluorescence and easily detectable alkaline phosphatase activity in the culture medium. The distribution of SEFP within the cells revealed that it was excluded from the nucleus, implying that the accumulation of SEFP in a secretory pathway, similar to that of the secretion signal-tagged FPs. Furthermore, the time- and dose-dependent release from the blockage of brefeldin A (BFA) confirmed that the secretion of SEFP was mediated by the secretion pathway and excluded leakage from viral infection. This SEFP reporter gene with traceable fluorescence and alkaline phosphatase activity may become a useful tool for studies on secretory protein production. PMID- 17401546 TI - Increased Factor V Leiden frequency is associated with venous thrombotic events among young Brazilian patients. AB - INTRODUCTION: Of the inherited thrombophilias, the Factor V Leiden (FVL) and the prothrombin mutant (FII G20210A) are associated with increased risk of venous thromboembolism (VTE). The C677T mutation of the methylenetetrahydrofolate reductase gene, which may lead to hyperhomocysteinemia, is also considered a risk factor for VTE in some studies. However, the frequency of these genetic risk factors may vary significantly among different populations. MATERIAL AND METHODS: The FVL, FII G20210A and C677T mutations were investigated by PCR-RFLP in 275 young VTE Brazilian patients as well as in 324 biologically unrelated individuals selected to compose the control group. RESULTS: The C677T mutation in the MTHFR gene was detected in 135 (49.1%) patients, of which 117 (42.5%) were identified as heterozygous and 18 (6.5%) as homozygous. The G20210A mutation was detected in 14 (5.1%) patients in heterozygosis. In both cases, no significant difference was observed when these results were compared to the frequencies observed in the control group. FVL was detected in heterozygosis in 19 (6.9%) patients, corresponding to a significantly increased frequency when compared to that observed for the control group (1.2%) (OR 5.9; 95% CI 2.08-16.79; p < 0.001). CONCLUSIONS: The data indicated that FVL is significantly associated with VTE among young Brazilian patients, but also supported previous evidence that VTE is a multi-factorial disease, resulting from the interaction of genetic and acquired risk factors. PMID- 17401547 TI - sICAM-1 measurements are independent of processing method and sampling site in patients with coronary artery disease. AB - sICAM-1 measurements are here shown to be independent of processing method (serum, platelet rich and platelet poor plasma) and sampling size (venous or arterial blood) in patients with coronary disease. PMID- 17401548 TI - Utilization of Fe3+-acinetoferrin analogs as an iron source by Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis, the causative agent of human tuberculosis, synthesizes and secretes siderophores in order to compete for iron (an essential micronutrient). Successful iron acquisition allows M. tuberculosis to survive and proliferate under the iron-deficient conditions encountered in the host. To examine structural determinants important for iron siderophore transport in this pathogen, the citrate-based siderophores petrobactin, acinetoferrin and various acinetoferrin homologs were synthesized and used as iron transport probes. Mutant strains of M. tuberculosis deficient in native siderophore synthesis or transport were utilized to better understand the mechanisms involved in iron delivery via the synthetic siderophores. Acinetoferrin and its derivatives, especially those containing a cyclic imide group, were able to deliver iron or gallium into M. tuberculosis which promoted or inhibited, respectively, the growth of this pathogen. PMID- 17401549 TI - [Urinary bladder augmentation using the stomach in patients with compensated renal insufficiency]. AB - This case report describes a female patient with bladder exstrophy and chronic compensated renal insufficiency as it relates to the topic of urinary bladder augmentation using the stomach and cutaneous vesicostomy with a Mitrofanoff continent stoma. Due to complete congenital bladder exstrophy the patient underwent functional reconstructive surgery for cystoschisis and several revision procedures. Because of a low-capacity, low-compliance bladder with bilateral urinary stasis in the kidneys, temporary percutaneous renal fistulas were created on both sides. After the patient had declined any follow-up during her pubertal years, advanced bilateral renal damage ensued. In a state of compensated renal insufficiency with creatinine levels >2 mg/dl, the patient was offered surgery for placement of an incontinent bladder substitute as an ileal or colon conduit because of the progressive kidney damage with obstructive uropathy. Since the patient rejected any form of wet stoma despite several consultations, after adequate preparations were met, a gastrocystoplasty and Mitrofanoff cutaneous vesicostomy with an umbilical stoma and artificial umbilicoplasty were performed. The patient catheterizes herself six times daily and is under close urological/nephrological supervision.In exceptional cases, gastrocystoplasty is an alternative to construction of a neobladder from bowel segments. It is particularly indicated for patients with compensated or already poor kidney function. The interventions are associated with several specific complications. There have as yet been no reports of malignant degeneration inside the gastric portion used for augmentation or the urothelium exposed to stomach acid. However, pathological and precancerous alterations of mucous membranes have been observed, which should be regularly monitored by endoscopy/biopsy and warrant further long term studies. PMID- 17401550 TI - Delirium assessment in the critically ill. AB - OBJECTIVE: To compare available instruments for assessing delirium in critically ill adults that have undergone validity testing and provide clinicians with strategies to incorporate these instruments into clinical practice. DESIGN: Medline (1966-September 2006) was searched using the key words: delirium, cognitive dysfunction, assessment, intensive care unit, and critical illness to identify assessment tools that have been used to evaluate delirium in critically ill adults. A special emphasis was placed on delirium assessment tools that have been properly validated. Data on how these tools have been adopted into clinical practice as well as strategies for clinicians to improve delirium assessment in the ICU are highlighted. MEASUREMENTS AND RESULTS: Six delirium assessment instruments including the Cognitive Test for Delirium (CTD), abbreviated CTD, Confusion Assessment Method-ICU, Intensive Care Delirium Screening Checklist, NEECHAM scale, and the Delirium Detection Score were identified. While each of these scales have undergone validation in critically ill adults, substantial differences exist among the scales in terms of the quality and extent of the validation effort, the specific components of the delirium syndrome each address, their ability to identify hypoactive delirium, their use in patients with a compromised level of consciousness, and their ease of use. CONCLUSIONS: Incorporation of delirium assessment into clinical practice in the intensive care unit using a validated tool may improve patient care. Clinicians can adopt a number of different strategies to overcome the many barriers associated with routine delirium assessment in the ICU. PMID- 17401551 TI - The contribution of prefrontal cortex to global perception. AB - Recent research suggests a role of top-down modulatory signals on perceptual processing, particularly for the integration of local elementary information to form a global holistic percept. In this study we investigated whether prefrontal cortex may be instrumental in this top-down modulation in humans. We measured detection thresholds for perceiving a circle defined by a closed chain of grating patches in 6 patients with prefrontal lesions, 4 control patients with temporal lesions and 17 healthy control subjects. Performance of patients with prefrontal lesions was worse than that of patients with temporal lesions and normal controls when the patterns were sparse, requiring integration across relatively extensive regions of space, but similar to the control groups for denser patterns. The results clearly implicate the prefrontal cortex in the process of integrating elementary features into a holistic global percept, when the elements do not form a "pop-out" display. PMID- 17401552 TI - Prospective dynamic balance control in healthy children and adults. AB - Balance control during gait initiation was studied using center of pressure (CoP) data from force plate measurements. Twenty-four participants were divided into four age groups: (1) 2-3 years, (2) 4-5 years, (3) 7-8 years, and (4) adults. Movement in the antero-posterior (CoPy) direction during the initial step was tau G analyzed, investigating the hypothesis that tau of the CoPy motion-gap (tau(CoPy)), i.e., the time it will take to close the gap at its current closure rate, is tau-coupled onto an intrinsic tau-G guide (tau(G)), by maintaining the relation tau(CoPy )= Ktau(G), for a constant K. Mean percentage of tau-guidance for all groups was >/=99%, resulting in all r(2) exceeding 0.95, justifying an investigation of the regression slope as an estimate of the coupling constant K in the tau-coupling equation. Mean K values decreased significantly with age and were for 2- to 3-year-olds 0.56, for 4- to 5-year-olds 0.50, for 7- to 8-year olds 0.47, and for adults 0.41. Therefore, the control of dynamic balance develops from the youngest children colliding with the boundaries of the base of support (K > 0.5) to the older children and adults making touch contact (K or =5 mm) in the neck were detected by ultrasonography and grossly later during necropsy. Histopathologic analysis revealed the cystic spaces to be lined by lymphatic endothelium supported by a connective tissue stroma. Control animals did not exhibit detectable lesions with either ultrasonography or necropsy. CONCLUSION: This model represents a promising tool for translational development of image guided interventions for lymphatic malformations. It may also serve as a model for the study of lymphangiogenesis and the development of anti-lymphangiogenic therapies. PMID- 17401559 TI - Molecular tools for analysis of gene function in parasitic microorganisms. AB - With the completion of several genome sequences for parasitic protozoa, research in molecular parasitology entered the "post-genomic" era. Accompanied by global transcriptome and proteome analysis, huge datasets have been generated that have added many novel candidates to the list of drug and vaccine targets. The challenge is now to validate these factors and to bring science back to the bench to perform a detailed characterization. In some parasites, like Trypanosoma brucei, high-throughput genetic screens have been established using RNA interference [for a detailed review, see Motyka and Englund (2004)]. In most protozoan parasites, however, more time-consuming approaches have to be employed to identify and characterize the function of promising candidates in detail. This review aims to summarize the status of molecular genetic tools available for a variety of protozoan pathogens and discuss how they can be implemented to advance our understanding of parasite biology. PMID- 17401560 TI - Vertical migration of aggregated aerobic and anaerobic ammonium oxidizers enhances oxygen uptake in a stagnant water layer. AB - Ammonium can be removed as dinitrogen gas by cooperating aerobic and anaerobic ammonium-oxidizing bacteria (AerAOB and AnAOB). The goal of this study was to verify putative mutual benefits for aggregated AerAOB and AnAOB in a stagnant freshwater environment. In an ammonium fed water column, the biological oxygen consumption rate was, on average, 76 kg O(2) ha(-1) day(-1). As the oxygen transfer rate of an abiotic control column was only 17 kg O(2) ha(-1) day(-1), biomass activity enhanced the oxygen transfer. Increasing the AnAOB gas production increased the oxygen consumption rate with more than 50% as a result of enhanced vertical movement of the biomass. The coupled decrease in dissolved oxygen concentration increased the diffusional oxygen transfer from the atmosphere in the water. Physically preventing the biomass from rising to the upper water layer instantaneously decreased oxygen and ammonium consumption and even led to the occurrence of some sulfate reduction. Floating of the biomass was further confirmed to be beneficial, as this allowed for the development of a higher AerAOB and AnAOB activity, compared to settled biomass. Overall, the results support mutual benefits for aggregated AerAOB and AnAOB, derived from the biomass uplifting effect of AnAOB gas production. PMID- 17401561 TI - Substrate specificity of rat brain neurolysin disclosed by molecular display system and putative substrates in rat tissues. AB - To search for the substrates, other than neurotensin, of rat brain neurolysin, a novel method of determining peptidase activity was developed using a yeast molecular display system. This is a useful and convenient method of handling homogenously pure proteins to evaluate the properties of neurolysin. The neurolysin gene was ligated to the C-terminal half of the alpha-agglutinin gene with a FLAG tag sequence and a yeast cell-surface molecular displaying plasmid was constructed. Display of neurolysin with correct folding and appropriate activity was verified by immunofluorescence staining and activity measurement of a bradykinin-related peptide. The cleavage sites of peptides were determined by high-performance liquid chromatography (HPLC) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The results showed the amino acid preferences of hydrophobic, aromatic, and basic residues, which were the same as those of soluble neurolysin. Moreover, this method clearly showed the presence of two recognition motifs in neurolysin. By using these motifs, novel substrate candidates of neurolysin in rat tissues were screened, and several bioactive peptides that regulate feeding were found. We also discussed the ubiquitous distribution of neurolysin in rat tissues and the functions of substrate candidate peptides. PMID- 17401562 TI - Synthesis of highly hydroxylated aromatics by evolved biphenyl dioxygenase and subsequent dihydrodiol dehydrogenase. AB - The evolved bphA1 (2049) gene, in which nine amino acids from the Pseudomonas pseudoalcaligenes KF707 BphA1 were changed to those from the Burkholderia xenovorans LB400 BphA1 (M247I, H255Q, V258I, G268A, D303E, -313G, S324T, V325I, and T376N), was expressed in Escherichia coli along with the bphA2A3A4 and bphB genes derived from strain KF707. This recombinant E. coli cells converted biphenyl and several heterocyclic aromatic compounds into the highly hydroxylated products such as biphenyl-2,3,2',3'-tetraol (from biphenyl), 2-(2,3 dihydroxyphenyl)benzoxazole-4,5-diol (from 2-phenylbenzoxazole), and 2-(2,5 dihydroxyphenyl)benzoxazole-4,5-diol [from 2-(2-hydroxyphenyl)benzoxazole]. The antioxidative activity of these generated compounds was markedly higher than that of the original substrate used. PMID- 17401563 TI - Production of medium-chain-length hydroxyalkanoic acids from Pseudomonas putida in pH stat. AB - Pseudomonas putida GP01 cells that had accumulated medium-chain-length polyhydroxyalkanoates (PHA(MCL)) secreted 3-hydroxyoctanoate and 3 hydroxyhexanoate when incubated in alkaline buffers. The release of acids strongly decreased the pH resulting in less efficient secretion of 3HA(MCL) at neutral pH. To increase the yield of secreted MCL-hydroxyalkanoates, experiments at constant pH in a pH stat apparatus were performed. High acid releasing rates were recorded for the wild type GP01 at pH 9.2 (0.60 mmol acid h(-1) g(-1) cellular dry weight [cdw]). At more alkaline constant pH values (pH 9.3-11), the initial acid secretion rates were even higher but rapidly decreased by time. When acid secretion of PHA depolymerase mutant GPo500 was tested (pH 9.2), considerably lower rates compared to wild type were recorded (0.18 mmol acid h( 1) g(-1) cdw). Determination of dissolved oxygen during acid release indicated different respiratoric activity in wild type (low) and mutant (high). Acid release of mutant, but not of the wild type, could be enhanced by aeration. Determination of PHA content of cells after alkaline incubation showed that the wild type had lost most of its accumulated PHA, whereas the PHA content of the depolymerase mutant was not significantly reduced. Considerable amounts of 3HA(MCL) were secreted by the wild type, but only little 3HA(MCL) were found for the depolymerase mutant. In summary, 3HA(MCL) can be more efficiently produced at constant high pH than by incubation without pH control. High PHA depolymerase activity enabled the wild type to compensate for the high external pH by secretion of PHA hydrolysis products, whereas production of protons at aerobic conditions presumably was responsible for the major portion of the observed acid releasing rates in the depolymerase mutant. PMID- 17401564 TI - Status of HFE mutation in thalassemia syndromes in north India. AB - Hereditary hemochromatosis is an autosomal recessive and most commonly inherited single gene disorder among Caucasians, with a prevalence of 5 per 1,000 and a carrier frequency of 1 in 10. Two point mutations were described and are referred as C282Y and H63D. In the present study, we have analyzed 729 north Indian samples for C282Y and H63D mutations. Of these, no allele of the C282Y mutation was seen, while 3 homozygous and 43 heterozygous for the H63D mutation were seen in the patients of thalassemia group. However, 47 cases were found heterozygous for the H63D mutation among the normal groups (11.16%). PMID- 17401565 TI - RNABC: forward kinematics to reduce all-atom steric clashes in RNA backbone. AB - Although accurate details in RNA structure are of great importance for understanding RNA function, the backbone conformation is difficult to determine, and most existing RNA structures show serious steric clashes (>or= 0.4 A overlap) when hydrogen atoms are taken into account. We have developed a program called RNABC (RNA Backbone Correction) that performs local perturbations to search for alternative conformations that avoid those steric clashes or other local geometry problems. Its input is an all-atom coordinate file for an RNA crystal structure (usually from the MolProbity web service), with problem areas specified. RNABC rebuilds a suite (the unit from sugar to sugar) by anchoring the phosphorus and base positions, which are clearest in crystallographic electron density, and reconstructing the other atoms using forward kinematics. Geometric parameters are constrained within user-specified tolerance of canonical or original values, and torsion angles are constrained to ranges defined through empirical database analyses. Several optimizations reduce the time required to search the many possible conformations. The output results are clustered and presented to the user, who can choose whether to accept one of the alternative conformations. Two test evaluations show the effectiveness of RNABC, first on the S-motifs from 42 RNA structures, and second on the worst problem suites (clusters of bad clashes, or serious sugar pucker outliers) in 25 unrelated RNA structures. Among the 101 S motifs, 88 had diagnosed problems, and RNABC produced clash-free conformations with acceptable geometry for 71 of those (about 80%). For the 154 worst problem suites, RNABC proposed alternative conformations for 72. All but 8 of those were judged acceptable after examining electron density (where available) and local conformation. Thus, even for these worst cases, nearly half the time RNABC suggested corrections suitable to initiate further crystallographic refinement. The program is available from http://kinemage.biochem.duke.edu . PMID- 17401566 TI - Outcome of patients with arthritis and parvovirus B19 DNA in synovial membranes. AB - To investigate the follow-up of the 17 patients during the period of 1995-2001 of the outpatient Clinic for Rheumatology at the University Hospital of Zurich with arthritis and the presence of parvovirus B19 DNA demonstrated by PCR in synovial biopsies. Seventeen patients of 163 with arthritis, which were routinely examined by needle arthroscopy during 1995-2001 with a positive parvovirus B19 DNA by PCR of synovial biopsy were reevaluated. Investigations included medical history, clinical examination and blood tests. Joint fluid was taken on patients with joint effusion. The observation period of the 17 patients (F:M = 11:6) was 2-8 years (O = 6.5 years). In 8 of 17 patients the arthritis could not be classified neither at entry nor during the follow up of the study. The arthritis could be diagnosed in six patients early in the onset of the disease and included three cases of lyme arthritis of the knee joint, two cases with arthritis following a gastrointestinal infection (one with Salmonella typhimurium--positive faecal test -and the other one with a culture negative agent), one patient probably had an infection-associated arthritis after a gastrointestinal infection with Entamoeba histolytica (Schirmer et al. in Rheumatol Int 18:37-38, 1998; Kasliwal in Am J Proctol Gastroenterol Colon Rectal Surg 32:12, 16, 28, 1981; Haslock and Wright in J R Coll Phys Lond 8:1554-162, 1974; Than-Saw et al. in Trop Geogr Med 44:355 358, 1992) with remission after antibiotic therapy. After a disease course of 9 months one patient could be classified as rheumatoid arthritis in the presence of anti-cyclic citrullinated antibodies but lack of rheumatoid factor. One patient with polyarthritis developed psoriasis of the skin 22 months later. From the nine patients with unclassified arthritis 4 (45%) got into complete remission with no symptoms or signs of joint inflammation after a disease course of 9-45 months, whereas 5 (55%) still demonstrate active non erosive arthritis (disease duration between 3 and 10 years). The presence of parvovirus B19 DNA in synovial tissue of patients with joint inflammation does not allow the diagnosis of parvovirus induced arthritis. If the arthritis remains unclassified and without erosions over time a virus associated aetiology may be assumed. However, no definitive diagnosis is possible even in the presence of parvovirus B19 DNA in synovial tissue. PMID- 17401567 TI - Prostaglandin E2 receptors EP1 and EP4 are up-regulated in rabbit chondrocytes by IL-1beta, but not by TNFalpha. AB - Prostaglandin E2 (PGE2) exerts its actions through the binding of the high affinity EP receptors. We wanted to evaluate the regulation of EP1 and EP4, and the expression of cyclooxygenase (COX)-2, main enzyme responsible for PGE2 synthesis in inflammatory situations, in healthy rabbit chondrocytes stimulated with inflammatory mediators locally increased during osteoarthritis. Articular chondrocytes obtained from healthy rabbits were stimulated with interleukin (IL) 1beta (0.1-100 u/ml) or tumour necrosis factor (TNF)alpha (100 ng/ml). Where indicated, cells were preincubated with non-steroidal antiinflammatory drugs (NSAIDs) (10(-6) M) to inhibit PGE2 synthesis. IL-1beta induced a dose and time dependent increase in EP1, EP4 and COX-2 expression. However, TNFalpha presence did not induce a significant modification in EP1, EP4 or COX-2 gene expression at any time of study. NSAID presence significantly inhibited PGE2 release but did not modify the EP receptors or COX-2 expression induced by IL-1beta. Our results indicate that EP1 and EP4 receptors, and COX-2 are up-regulated in IL-1beta stimulated chondrocytes, while no significant modifications are observed in TNFalpha-stimulated cells. NSAIDs were unable to modify the expression of these mediators induced by IL-1beta. Therefore, the increase in PGE2 synthesis, induced by IL-1beta, does not seem to mediate the increase in EP receptor expression, in rabbit chondrocytes. PMID- 17401568 TI - An ovarian tumour with a potential appendiceal origin. AB - A 14-year-old girl underwent left oopherectomy for a multicystic ovarian mass. Histology revealed this to be an intestinal type mucinous borderline tumour (IMBT). In view of this, a semi-urgent laparoscopic appendicectomy was carried out. The appendix was histologically normal. IMBT of the ovary is a mucinous tumour with atypical proliferation of the goblet cell containing epithelium. It is known to occur simultaneously with tumours of the appendix. Paediatric surgeons need to be aware of this rare tumour and when operating on any ovarian pathology should always inspect the peritoneal cavity for mucinous deposits and examine the appendix. PMID- 17401569 TI - [Painful and swollen ankles: no arthritis]. AB - A 53 year old male patient suffering from painful and swollen ankles showed a large enhancement on skeletal scintigraphy. The DXA bone density values for the neck of the femur and the lumbar vertebrae indicated severe osteoporosis. The conventional diagnostic x-ray revealed stress fractures of both distal tibiae (Looser's transformation zones). The patient had been treated with glucocorticoids for many years because of a liver transplantation. A further risk factor was impaired renal function with a hypovitaminosis D3 and secondary hyperparathyroidism. PMID- 17401570 TI - Silver diagnosis in neuropathology: principles, practice and revised interpretation. AB - Silver-staining methods are helpful for histological identification of pathological deposits. In spite of some ambiguities regarding their mechanism and interpretation, they are widely used for histopathological diagnosis. In this review, four major silver-staining methods, modified Bielschowsky, Bodian, Gallyas (GAL) and Campbell-Switzer (CS) methods, are outlined with respect to their principles, basic protocols and interpretations, thereby providing neuropathologists, technicians and neuroscientists with a common basis for comparing findings and identifying the issues that still need to be clarified. Some consider "argyrophilia" to be a homogeneous phenomenon irrespective of the lesion and the method. Thus, they seek to explain the differences among the methods by pointing to their different sensitivities in detecting lesions (quantitative difference). Comparative studies, however, have demonstrated that argyrophilia is heterogeneous and dependent not only on the method but also on the lesion (qualitative difference). Each staining method has its own lesion dependent specificity and, within this specificity, its own sensitivity. This "method- and lesion-dependent" nature of argyrophilia enables operational sorting of disease-specific lesions based on their silver-staining profiles, which may potentially represent some disease-specific aspects. Furthermore, comparisons between immunohistochemical and biochemical data have revealed an empirical correlation between GAL+/CS-deposits and 4-repeat (4R) tau (corticobasal degeneration, progressive supranuclear palsy and argyrophilic grains) and its complementary reversal between GAL-/CS+deposits and 3-repeat (3R) tau (Pick bodies). Deposits containing both 3R and 4R tau (neurofibrillary tangles of Alzheimer type) are GAL+/CS+. Although no molecular explanations, other than these empiric correlations, are currently available, these distinctive features, especially when combined with immunohistochemistry, are useful because silver staining methods and immunoreactions are complementary to each other. PMID- 17401571 TI - Minimally invasive plating for complex humeral shaft fractures. AB - INTRODUCTION: The treatment for humeral shaft fracture is still controversial, especially for complex fractures. The purpose of this study was to evaluate the clinical and radiographic outcomes of treating complex humeral shaft fractures by using locking compression plate with minimally invasive technique. MATERIALS AND METHODS: A total of 21 patients with 21 acute complex humeral shaft fractures (AO type C) were included in this study. All were treated with locking compression plate in a minimally invasive way. The average age of the patients was 42.9 years and there were 14 males and 7 females. Plain radiographs of humerus were used to assess bony union. Functional recovery of the shoulder joint was assessed using the Constant and HSS scoring systems. RESULTS: The patients were followed for a mean of 28.7 months (range 19-37). With one operation, nineteen fractures (90.4%) achieved a solid union in an average of 14.3 weeks. At final follow-up, 20 patients (95.2%) had satisfactory alignment, except for one associated with olecranon fracture, all patients had a good to excellent elbow function with a mean HSS score of 91.7 points. Eighteen patients (85.7%) achieved satisfactory shoulder function with a mean Constant score of 83.1 points. One superficial infection (4.8%) and two non-unions (9.6%) were found postoperatively. CONCLUSIONS: It was concluded that use of a minimally invasive plate osteosynthesis with locking compression plate is a safe alternative way to classic surgical methods in the treatment of complex humeral shaft fractures, which had a high union rate with less risk of complications. PMID- 17401572 TI - Palisade cartilage tympanoplasty for management of subtotal perforations: a comparison with the temporalis fascia technique. AB - The use of cartilage as a grafting material has been advocated in cases at high risk for failure, such as subtotal perforations, adhesive processes, tympanosclerosis and residual defects after primary tympanoplasties. The cartilage palisade technique, in which the TM is fully reconstructed with palisade-shaped cartilage pieces, was first described by Heermann. Cartilage material has been criticized because of concerns regarding hearing results, however, large cartilage plates with thicknesses 50% of the whole TM area), intact ossicular chain, at least more than 1 month dry period and normal middle ear mucosa were included in the study. Palisade cartilage tympanoplasty group included 23 cases, whereas 28 patients were were in the fascia group. INTERVENTION: therapeutic. Graft take rates were noted in 51 patients and pre- and post operative audiograms were compared using the chi square test with Yates' continuity correction, Fisher's exact test, t test and repeated measures ANOVA. Graft take was achieved in 22 patients (95.7%) in palisade cartilage group and in 21 patients (75%) in temporalis fascia group (P = 0.059). Mean speech reception threshold, air-bone gap and pure-tone average scores comparing the gain between both techniques showed no significant changes in the threshold (P > 0.05). However the functional success was achieved with the palisade cartilage technique postoperatively, regarding to mean air-bone gap and speech reception threshold changes. Our experience with palisade cartilage technique demonstrates that subtotal or total perforations at high risk for graft failure, can be treated efficiently and a durable and resistant reconstruction of the TM with reasonable hearing can be achieved. PMID- 17401573 TI - Feet rolled over by cars: radiological and histological considerations from experiments. AB - This study investigates the question of whether bone structures are injured when a vehicle rolls over a foot. A total of 15 detached feet from deceased persons who had donated their bodies to research were rolled over using a VW Passat station wagon. The feet were enclosed in various types of shoes. The front left tire of the vehicle, inflated to 1.8 bar and driven at walking speed, ran over the feet at a right angle to the long axis. The feet were dissected, and histological and radiological examinations were carried out. The only macroscopically well-defined abrasions of the epidermis were on the back of the foot in the area of contact with the tire and only where the foot had not been covered by a shoe. These abrasions were also well presented histologically. No injuries to the bone structures of the feet, in the form of incomplete fractures, corticalis interruptions or spongiosa compressions were ascertained, either radiologically or microradiologically. PMID- 17401574 TI - Comparative analysis of the applicability of the skeletal age determination methods of Greulich-Pyle and Thiemann-Nitz for forensic age estimation in living subjects. AB - Evaluation of hand ossification is a main pillar of radiological age diagnosis in living subjects. In the present study, we comparatively assessed the Greulich Pyle and Thiemann-Nitz methods for accuracy of age estimation and degree of acceleration in the respective reference populations. For this, the skeletal age of 649 hand X-rays from German subjects aged 1-18 years was determined by both methods. Accuracy of the age estimates was determined based on regression and measures of certainty. In terms of accuracy, both methods seem to be equally well suited for forensic age diagnostics. The degree of acceleration in the reference populations for the two methods was calculated as the mean difference between the estimated skeletal age and the actual age of a test subject. Compared to the Greulich-Pyle population, the Thiemann-Nitz population was accelerated by 0.44 years in both male and female subjects. When an expert opinion is required and one cannot exclude the possibility that the investigated subject may come from a population with a high acceleration status, the Thiemann-Nitz method should preferably be used to prevent overestimation of age. PMID- 17401575 TI - Lengthening contractions differentially affect p70s6k phosphorylation compared to isometric contractions in rat skeletal muscle. AB - The purpose of this investigation was to determine if p70(s6k) phosphorylation is dependent on the mode of resistance exercise (e.g. isometric vs. lengthening). Two groups (n = 5 each) of Female Sprague Dawley rats, approximately 12 weeks old, were tested. Rats were anesthetized and indwelling electrodes used to stimulate the right hind limb muscles via the sciatic nerve. The tibialis anterior (TA) muscle of Group 1 rats were exposed to three sets of ten isometric resistance contractions while the TA of Group 2 rats were exposed to three sets of ten resistance contractions that involved lengthening. Contralateral TA muscles served as non-exercised controls. The TA muscle was harvested 6 h post exercise and then the rat was euthanized. Muscle samples were processed to compare p70(s6k) phosphorylation between groups. A single bout of TA contractions that involved muscle lengthening resulted in significantly (p < 0.05) higher levels of phospho-p70(s6k) six hours post exercise compared to controls and isometric contractions. The differences in total p70(s6k) six hours post exercise were not significantly different between groups. Results suggest that signal transduction pathways activated by isometric exercise may differ (i.e., a non p70(s6k) activation pathway) from that activated by lengthening exercise. PMID- 17401576 TI - Interaction of syntaxin with a single Kv1.1 channel: a possible mechanism for modulating neuronal excitability. AB - Voltage-gated K(+) channels are crucial for intrinsic neuronal plasticity and present a target for modulations by protein-protein interactions, notably, by exocytotic proteins demonstrated by us in several systems. Here, we investigated the interaction of a single Kv1.1 channel with syntaxin 1A. Syntaxin decreased the unitary conductance of all conductance states (two subconductances and a full conductance) and decreased their open probabilities by prolongation of mean closed dwell-times at depolarized potentials. However, at subthreshold potentials syntaxin 1A increased the probabilities of the subconductance states. Consequently, the macroscopic conductance is decreased at potentials above threshold and increased at threshold potentials. Numerical modeling based on steady-state and kinetic analyses suggests: (1) a mechanism whereby syntaxin controls activation gating by forcing the conductance pathway only via a sequence of discrete steps through the subconductance states, possibly via a breakdown of cooperative movements of voltage sensors that exist in Kv1.1; (2) a physiological effect, apparently paradoxical for an agent that reduces K(+) current, of attenuating neuronal firing frequency via an increase in K(+) shunting conductance. Such modulation of the gain of neuronal output in response to different levels of syntaxin is in accord with the suggested role for Kv1.1 in axonal excitability and synaptic efficacy. PMID- 17401577 TI - Moebius-Poland syndrome and hypogonadotropic hypogonadism. PMID- 17401578 TI - Recovery kinetics of heart rate and oxygen uptake in long-term survivors of acute leukemia in childhood. AB - Analysis of the recovery period following physical exercise has gained importance in evaluating cardiopulmonary capacity, not only in athletes but also in patients with proven or suspected heart failure. The purpose of this study was to apply these methods to long-term survivors of acute lymphoblastic leukemia (ALL) in childhood, who are at risk of developing anthracycline-induced cardiomyopathy. Nine children (mean age 12 years) and 10 adults (mean age 24 years) were included in the study after treatment for childhood ALL. Recovery of oxygen uptake and heart rate following maximal spiroergometric exercise was compared to that in 29 trained and untrained age-matched controls. The change in oxygen uptake (DeltaVO2) and heart rate (DeltaHR) between maximal effort and 60 s of recovery did not differ significantly, either between children after oncological therapy (DeltaVO2: 14.95 ml/kg, DeltaHR: 35 bpm) and healthy children (DeltaVO2: 15.85 ml/kg, DeltaHR: 37 bpm), or between adult former oncological patients (DeltaVO2: 13.1 ml/kg, DeltaHR: 27 bpm) and untrained adults (DeltaVO2: 15.7 ml/kg, DeltaHR: 31 bpm). There was, however, a significant difference in DeltaVO2 between trained adults (DeltaVO2: 24.5 ml/kg) and both untrained adult controls (DeltaVO2: 15.7 ml/kg, p=0.004) and adult patients (DeltaVO2: 13.1 ml/kg, p=0.0002). This difference was not detected for heart rate. In conclusion, the recovery period did not reveal a discernible difference in cardiopulmonary capacity between former ALL patients and untrained age-matched controls. We did confirm that heart rate and oxygen uptake recovery serve as indicators of physical fitness. PMID- 17401579 TI - Ultrastructure of spermiogenesis and the spermatozoon of Paramphistomum microbothrium [Fischoeder 1901] (Digenea: Paramphistomidae), a parasite of Bos taurus in Senegal. AB - The ultrastructural study of spermiogenesis and the spermatozoon of Paramphistomum microbothrium gave evidence of some characteristics of this digenean. The intercentriolar body located between two striated roots and two centrioles exhibits a symmetric plane. The two external bands of this intercentriolar body are made up of a line of granules. The striated roots are about 3.7 microm long. During spermiogenesis, a flagellar rotation of 90 degrees is described. The spermatozoon is characterized in its anterior part by the existence of external ornamentations and a lateral expansion exhibiting a "spine like body". Such a spine-like body has never been described before in a digenean. The region II of the spermatozoon is asymmetrical and also characterized by the absence of mitochondrion. The posterior end of this spermatozoon exhibits a nucleus and a few microtubules. PMID- 17401580 TI - Strongyloides stercoralis hyper-infection syndrome in HIV+/AIDS patients in Iran. AB - Strongyloides stercoralis is an intestinal nematode in humans, distributed through tropical and subtropical regions of the world. In most individuals, the infection has a chronic nature due to auto-infection at the low level. Accelerated auto-infection, mainly after an alteration in immune status, can cause a syndrome of severe hyper-infection or potentially fatal disseminated strongyloidiasis. Due to the increasing numbers of immunocompromised patients in Iran, strongyloidiasis is an emerging public health concern in the country. In the current study, which was carried out between 2003 and 2005, for the investigation on strongyloidiasis in HIV(+)/AIDS patients, a total of 781 patients were examined by agar plate culture, formalin ether concentration, and direct smear preparation of stool samples. According to the results, 2 out of 781 HIV( + )/AIDS patients were found infected with S. stercoralis, but both patients were at the progressive stage of AIDS and showing severe hyper-infection syndrome. In both cases, numerous rhabditiform and filariform larvae were found in fresh stool direct smears, and rapid and intensive development of parasite in agar plate cultures. In conclusion, in the progressive stages of AIDS, as a result of immunosuppression conditions or in the context of chemotherapy, S. stercoralis is capable of inducing overwhelming infection. PMID- 17401581 TI - Abundance matters: a field experiment testing the more individuals hypothesis for richness-productivity relationships. AB - The more individuals hypothesis (MIH) postulates that productivity increases species richness by increasing mean equilibrium population size, thereby reducing the probability of local extinction. We tested the MIH for invertebrates colonizing microcosms that simulated tree holes by manipulating productivity through additions of leaf or animal detritus and subsequently determining the relationships among richness, total abundance, abundance per species, and measures of productivity. We quantified productivity as the rate of microorganism protein synthesis, microorganism metabolic rate, nutrient ion concentration, and type and amount of detritus. Microcosms with animal detritus attracted more species, more individuals per species, and more total individuals than did microcosms with similar amounts of leaf detritus. Relationships between richness or abundance and productivity varied with date. Richness in June increased as a linear function of productivity, whereas the power function predicted by the MIH fit best in July. Abundance in June and July was best described by a power function of productivity, but the linear function predicted by the MIH fit best in September. Abundance per species was best described by a power function of productivity in June and July. Path analysis showed that the indirect effect of productivity through abundance on richness that is predicted by MIH was important in all months, and that direct links between productivity and richness were unnecessary. Our results support many of the predictions of the MIH, but they also suggest that the effects of abundance on richness may be more complex than expected. PMID- 17401582 TI - Isotope fractionation and 13C enrichment in soil profiles during the decomposition of soil organic matter. AB - The mechanisms behind the (13)C enrichment of organic matter with increasing soil depth in forests are unclear. To determine if (13)C discrimination during respiration could contribute to this pattern, we compared delta(13)C signatures of respired CO(2) from sieved mineral soil, litter layer and litterfall with measurements of delta(13)C and delta(15)N of mineral soil, litter layer, litterfall, roots and fungal mycelia sampled from a 68-year-old Norway spruce forest stand planted on previously cultivated land. Because the land was subjected to ploughing before establishment of the forest stand, shifts in delta(13)C in the top 20 cm reflect processes that have been active since the beginning of the reforestation process. As (13)C-depleted organic matter accumulated in the upper soil, a 1.0 per thousand delta(13)C gradient from -28.5 per thousand in the litter layer to -27.6 per thousand at a depth of 2-6 cm was formed. This can be explained by the 1 per thousand drop in delta(13)C of atmospheric CO(2) since the beginning of reforestation together with the mixing of new C (forest) and old C (farmland). However, the isotopic change of the atmospheric CO(2) explains only a portion of the additional 1.0 per thousand increase in delta(13)C below a depth of 20 cm. The delta(13)C of the respired CO(2) was similar to that of the organic matter in the upper soil layers but became increasingly (13)C enriched with depth, up to 2.5 per thousand relative to the organic matter. We hypothesise that this (13)C enrichment of the CO(2) as well as the residual increase in delta(13)C of the organic matter below a soil depth of 20 cm results from the increased contribution of (13)C-enriched microbially derived C with depth. Our results suggest that (13)C discrimination during microbial respiration does not contribute to the (13)C enrichment of organic matter in soils. We therefore recommend that these results should be taken into consideration when natural variations in delta(13)C of respired CO(2) are used to separate different components of soil respiration or ecosystem respiration. PMID- 17401583 TI - The importance of parasite life history and host density in predicting the impact of infections in red deer. AB - We studied a macroparasite (Elaphostrongylus cervi, Nematoda) and a microparasite infection (tuberculosis, TB) in red deer (Cervus elaphus) across different populations where managers manipulated host condition, density and aggregation by providing supplemental food. We aimed to test whether and, if so, how persistence and transmission of both parasites differentially varied as host body condition and population density varied. We took account of sex, as red deer life history greatly concerns sex-related traits. Changes in host factors had different consequences for the spread of each parasite type. Individual presence of tuberculosis was positively associated with host density, whereas E. cervi abundance negatively related to host density and enhanced body condition. There was lack of body condition density dependence; and body condition was mainly dependent on the amount of supplemental food provided, but also on habitat quality descriptors. Overall, our results suggest that body condition was improved at the cost of increased host contact rates, which implied an ecological trade-off between acquiring resources to cope with E. cervi, a macroparasite, and concurrent exposure to mycobacteria. By the simultaneous study of both infections, this research suggests that the effects of changes in host number and population structure on disease spread and persistence need to take into account variation in life histories of the parasites. These findings also raise concern about the ecological consequences of diseases and management of wildlife on host life history. PMID- 17401584 TI - Growth of prepubertal children on dialysis. AB - Growth failure is a common and significant clinical problem for children on dialysis and often remains a major impediment to their rehabilitation. Early referral to a paediatric nephrology centre and appropriate management before the initiation of dialysis may significantly prevent growth deterioration. Growth in children on dialysis can be affected by nutritional, metabolic, and hormonal changes. Early diagnosis of malnutrition and aggressive management should be a priority. Gastrostomy feeding should be used when adequate oral intake to maintain normal height and weight velocity cannot be achieved. Active vitamin D metabolites should be used carefully, to prevent low-turnover bone disease. All children should have an adequate regimen of dialysis and an appropriate management of malnutrition, renal osteodystrophy, metabolic acidosis, salt wasting and anaemia, before recombinant human growth hormone (rhGH) administration is considered. The current challenge of reversing growth impairment in children on dialysis can only be achieved by optimization of their care. PMID- 17401585 TI - Sonographic assessment of normal kidney dimensions in the first year of life--a study of 992 healthy infants. AB - The omission of standards for renal length in infants younger than 1 year may result in a statistically significant increase in the frequency of "spurious" nephromegaly. Nonetheless, there are only a few reports specifically dealing with normal kidney dimensions in infants. Based on sonographic assessments performed on a sample of 992 healthy infants, between January 2002 and December 2004, this paper sets up standards for normal kidney dimensions in children aged 0-3 months, 3-6 months, 6-9 months, and 9-12 months and establishes correlations between kidney dimensions (length, width, and volume) and body length and weight. Linear as well as non-linear nomograms, with percentiles for all the kidney variables examined, based on body length, are provided. Also, statistically significant differences in mean values (P=0.000) for all the observed neonatal kidney parameters, depending on gestational age at birth, are demonstrated. Principal advantages of our nomograms are that they are based on a large number of examined healthy infants and that kidney dimensions are related to body length. In addition, subjects are divided into four sub-annual age groups demonstrating gender-related differences in renal growth dynamics. Our linear nomograms are easier to use for routine clinical practice, but the percentile-based non-linear nomograms we present cover a much wider range of variations in normal infant kidney dimensions. PMID- 17401586 TI - A novel mutation in KCNJ1 in a Bartter syndrome case diagnosed as pseudohypoaldosteronism. AB - Bartter syndrome (BS) is a genetic disorder with hypokalemic metabolic alkalosis and is classified into five types. One of these, type II BS (OMIM 241200), is classified as neonatal Bartter syndrome, which is caused by mutations in the KCNJ1 gene. Transient hyperkalemia and hyponatremia are usually noted in the early postnatal period, but as type II BS is a relatively rare disease, its exact clinical course and genetic background have not yet been thoroughly characterized. This report concerns a male type II BS patient with a novel mutation in the KCNJ1 gene. The unique clinical findings of this case are that hyperkalemia (8.9 mEq/l), hyponatremia, and metabolic acidosis detected in the early postnatal period led to a diagnosis of pseudohypoaldosteronism (PHA). As an adolescent, however, the patient currently shows normal potassium levels and normal renal function, although with hypercalciuria and nephrocalcinosis, without having received any treatment. In such cases, KCNJ1 mutations should be suspected. In our case, genetic analysis of the KCNJ1 gene identified a novel homozygous 1-bp deletion mutation (c.607 del. C in exon 5). PMID- 17401587 TI - Renal transplant in methylmalonic acidemia: could it be the best option? Report on a case at 10 years and review of the literature. AB - Methylmalonic acidemia (MMA) is an inborn error of organic acid metabolism. Patients with severe disease develop many complications despite treatment; often, the disease progresses to severe damage of the central nervous system or to end stage renal disease (ESRD). When medical treatment is ineffective, liver, kidney, or combined liver and kidney transplantation is advocated. At present, there are no definite guidelines as for the organ to be transplanted, and results are inconsistent. We report on a 27-year-old woman with MMA MUT0. The clinical symptoms developed at age 4 months. She progressed to ESRD and received a kidney transplant in November 1996 at age 17 years. One hundred and twenty months after transplant, renal function is normal; although urinary levels of methylmalonic acid are above normal limits, no episodes of metabolic decompensation have been observed after transplantation. Although liver is the major site of methylmalonyl CoA mutase activity, this case and similar ones in the literature suggest that the smaller mutase activity present in the transplanted kidney may be sufficient to ensure partial correction of the metabolism of organic acids sufficient to prevent the onset of episodes of metabolic decompensation. It is worth investigating whether kidney transplant can be a safer and more satisfactory alternative to liver transplantation in cases of MMA unresponsive to medical treatment although urine MMA excretion remains significantly elevated. PMID- 17401588 TI - Clinical study on the primary stability of two dental implant systems with resonance frequency analysis. AB - Primary stability has a major impact on the long-term success of dental implants. The aim of this study was to investigate the correlation of resonance frequency analysis (RFA) and insertion torque of self-tapping and non-self-tapping implants and their respective differences in primary stability. A group of 263 patients were treated with a total of 602 conically formed dental implants: 408 non-self tapping Ankylos and 194 self-tapping Camlog. The maximum insertion torque during implant placement was recorded. Resonance frequency, measured as the implant stability quotient (ISQ), was assessed once immediately after insertion and twice 3 months later. Torque values of the non-self-tapping implants were significantly higher than those in the self-tapping group (p = 0.023). RFA did not show differences between the 2 groups (p = 0.956), but a correlation between ISQ values after implantation and 3 months after implant placement was measured (r = 0.712). Within the implant systems, no correlation between insertion torque and resonance frequency values could be determined (r = 0.305). Our study indicates that the ISQ values obtained from different implant systems are not comparable. The RFA does not appear suitable for the evaluation of implant stability when used as a single method. Higher insertion torque of the non-self-tapping implants appeared to confirm higher clinical primary stability. PMID- 17401589 TI - Salivary constituents and acidogenic microbial counts in coronary artery bypass graft patients from baseline to three-years after operation. AB - Data on saliva in coronary artery bypass graft (CABG) surgery patients are sparse. Understanding salivary parameters, however, may aid clinical decision making. We hypothesized that cardiac surgery might affect patients' salivary flow rates and buffering, salivary proteins, and microbial counts. A 3-year, open follow-up study was conducted examining salivary flow, its chemical composition, and acidogenic microbial counts in 89 CABG surgery patients. The changes in salivary flow and proteins between baseline and 3-year post-CABG surgery were assessed using paired t-test and, with respect to the median of number of drugs used daily, by use of a nonparametric rank sum test. The results showed no long term change in salivary flow rates and buffering capacity. With the exception of salivary urea, IgA and IgM concentration, and lysozyme output, the differences in salivary proteins between baseline and 3-year post-CABG were not statistically significant. No difference was observed in saliva values between patients taking drugs below or above the median number of drugs. Acidogenic microbial counts remained the same throughout the study. In conclusion, the salivary flow rates and constituents did not practically change in patients who underwent CABG surgery during the 3-year follow-up. PMID- 17401590 TI - [The importance of dental-based treatment shown on the case report of a pontine abscess caused by Streptococcus viridans]. AB - BACKGROUND: The treatment of infectious oral lesions is an important prophylactic procedure preceding immunsuppressive therapy. The case reported underlines the importance of this treatment by showing severe complications of an infective dental focus in an immunocompetent patient. CASE REPORT: A 65 year old patient was referred to hospital due to neurological disorders including hypaesthesia, movement deficits, nausea and falls. Because of progressive aggravation with hemiplegia a craniotomy and biopsy of a suspicious area in the pons was performed. A brain stem abscess caused by Streptococcus viridans was diagnosed. Brain stem abscesses are very rare events who contribute to only about 0.6%-6% of all intracranial abscesses. The intraoral inspection revealed a periodontally diseased and carious affected dentition with many missing teeth. Other foci were ruled out. Because of persistency of the symptoms another operation with abscess drainage was performed. Afterwards the clinical symptoms improved but hemiplegia was still present. CONCLUSIONS: Even minor dental procedures as well as existence of a carious or periodontal disease may cause bacteremia with severe septical implications. Patients at risk (e.g. dieases of the heart valves or diabetes) should always treated with an antibiotic prophylaxis before treatment according to the guidelines of the DGZMK. All oro-pharyngeal infectious foci have to be treated sufficiently to prevent sceptical complications. Even more important is the prophylactic treatment preceding chemo- or radiation therapy. PMID- 17401591 TI - Q fever osteoarticular infection: four new cases and a review of the literature. AB - Q fever is a worldwide-occurring zoonosis caused by Coxiella burnetii. Better knowledge of the disease and of evolving diagnostics can enable recognition of unusual manifestations. Reported here are four cases of Q fever osteoarticular infections in adults: two cases of Q fever tenosynovitis, which represent the first two reports of this infection, and two cases of Q fever spondylodiscitis complicated by paravertebral abscess. In addition, the literature is reviewed on the 15 previously reported cases of Q fever osteoarticular infection, six of which were vertebral infections. Osteomyelitis is the usual manifestation Q fever osteoarticular infection. Because its onset is frequently insidious, diagnosis is usually delayed. The main differential diagnosis is mycobacterial infection, based on the histological granulomatous presentation of lesions. Whereas serology is the reference diagnostic method for Q fever, detection of C. burnetii in tissue specimens by PCR and cell culture provides useful additional evidence of infection. Culture-negative osteoarticular samples with granulomatous presentation upon histological examination should raise suspicion of Q fever. PMID- 17401592 TI - Economic aspects of the new Spanish laws on pharmaceutical preparations. AB - In this article, we provide readers with a summary of the main economic features of the recently approved Spanish law regarding pharmaceutical preparations entitled the Law of Guarantees and Rational Use of Pharmaceuticals and Health Products (Law 29/2006 of July 26th). We review information on pricing, reference pricing, promotion of generic drugs, public reimbursement, patients' contributions, and penalties. Other aspects of minor economic relevance such as the information on prices shown in the packs and the transparency on the results of clinical trials irrespective of whether these are positive or not, are not addressed in this paper. We conclude with some observations on the new horizons opened up by this new legal framework. We have not included any detailed references to the new law; we merely wish to comment on certain aspects of its application. PMID- 17401593 TI - Factors influencing hospital costs of lung cancer patients in Northern Ireland. AB - Lung cancer is a major cause of morbidity and mortality. In this paper, the hospital costs incurred by 724 lung cancer patients diagnosed in 2001 were determined by review of case notes. These represented all patients diagnosed with lung cancer in Northern Ireland on whom data existed in that year. Total hospital costs in the 12 months from presentation for the 724 patients were 3.99 million pounds. Average patient costs were 5,956 pounds for patients diagnosed with non small cell lung cancer and 5,876 pounds for those with small cell lung cancer. The main component of cost was inpatient stay, representing between 62 and 84% of costs depending on cell type. Multivariate analyses revealed significant differences in cost related to staging, co-morbidities, age, and deprivation. Total annual hospital costs were 13 times as high as the estimated enforcement cost of the smoke-free legislation in Northern Ireland. PMID- 17401594 TI - Supplier-induced demand: re-examining identification and misspecification in cross-sectional analysis. AB - This paper re-examines criticisms of cross-sectional methods used to test for supplier-induced demand (SID) and re-evaluates the empirical evidence using data from Australian medical services. Cross-sectional studies of SID have been criticised on two grounds. First, and most important, the inclusion of the doctor supply in the demand equation leads to an identification problem. This criticism is shown to be invalid, as the doctor supply variable is stochastic and depends upon a variety of other variables including the desirability of the location. Second, cross-sectional studies of SID fail diagnostic tests and produce artefactual findings due to model misspecification. Contrary to this, the re evaluation of cross-sectional Australian data indicate that demand equations that do not include the doctor supply are misspecified. Empirical evidence from the re evaluation of Australian medical services data supports the notion of SID. Demand and supply equations are well specified and have very good explanatory power. The demand equation is identified and the desirability of a location is an important predictor of the doctor supply. Results show an average price elasticity of demand of 0.22 and an average elasticity of demand with respect to the doctor supply of 0.46, with the impact of SID becoming stronger as the doctor supply rises. The conclusion we draw from this paper is that two of the main criticisms of the empirical evidence supporting the SID hypothesis have been inappropriately levelled at the methods used. More importantly, SID provides a satisfactory, and robust, explanation of the empirical data on the demand for medical services in Australia. PMID- 17401595 TI - Quantitative monitoring of extracellular matrix production in bone implants by 13C and 31P solid-state nuclear magnetic resonance spectroscopy. AB - We used (31)P and (13)C solid-state nuclear magnetic resonance (NMR) spectroscopy to detect and analyze the major organic and inorganic components (collagen type I and bioapatite) in natural rabbit bone and beta-tricalcium phosphate implants loaded with osteogenically differentiated mesenchymal stem cells. High-resolution solid-state NMR spectra were obtained using the magic-angle spinning (MAS) technique. The (31)P NMR spectra of bone specimens showed a single line characteristic of bone calcium phosphate. (13)C cross-polarization (CP) MAS NMR spectra of bone exhibited the characteristic signatures of collagen type I with good resolution for all major amino acids in collagen. Quantitative measurements of (13)C-(1)H dipolar couplings indicated that the collagen segments are very rigid, undergoing only small amplitude fluctuations with correlation times in the nanosecond range. In contrast, directly polarized (13)C MAS NMR spectra of rabbit bone were dominated by signals of highly mobile triglycerides. These quantitative investigations of natural bone may provide the basis for a quality control of various osteoinductive bone substitutes. We studied the formation of extracellular bone matrix in artificial mesenchymal stem cell-loaded beta tricalcium phosphate matrices that were implanted into the femoral condyle of rabbits. The NMR spectra of these bone grafts were acquired 3 months after implantation. In the (31)P NMR spectra, beta-tricalcium phosphate and bone calcium phosphate could be distinguished quantitatively, allowing recording of the formation of the natural bone matrix. Further, (13)C CPMAS allowed detection of collagen type I that had been produced in the implants. Comparison with the spectroscopic data from natural bone allowed assessment of the quality of the bone substitute material. PMID- 17401596 TI - Evaluation of adherence, hydrophobicity, aggregation, and biofilm development of Flavobacterium johnsoniae-like isolates. AB - Flavobacterium spp. isolates have been identified in diverse biofilm structures, but the mechanism of adherence has not been elucidated. The absence of conventional biofilm-associated structures such as fimbriae, pili, and flagella suggest that surface hydrophobicity, and/or autoaggregation and coaggregation may play an important role in adherence and biofilm formation. The biofilm-forming capacity of 29 Flavobacterium johnsoniae-like isolates obtained from South African aquaculture systems was assessed using microtiter plate assays. The role of hydrophobicity [salting aggregation test (SAT) and bacterial adherence to hydrocarbons (BATH) assays], autoaggregation, and coaggregation on biofilm formation by Flavobacterium spp. was also investigated, while biofilm structure was examined using flow cells and microscopy. All isolates displayed a hydrophilic nature, but showed varying levels of adherence in microtiter assays. Significant negative correlations were observed between adherence and biofilm forming capacity in nutrient-poor medium at 26 degrees C and BATH hydrophobicity and motility, respectively. Isolates displayed strain-to-strain variation in their autoaggregation indices and their abilities to coaggregate with various Gram-negative and Gram-positive organisms. Microcolony and/or biofilm development were observed microscopically, and flavobacterial isolates displayed stronger biofilm structures and interaction with a Vibrio spp. isolate than with an Aeromonas hydrophila isolate. The role of extracellular polysaccharides and specific outer membrane proteins will have to be examined to reveal mechanisms of adherence and coaggregation employed by biofilm-forming F. johnsoniae-like strains. PMID- 17401597 TI - Relationships between soil organic status and microbial community density and genetic structure in two agricultural soils submitted to various types of organic management. AB - The effects of soil organic management on indigenous microorganisms were studied by comparing mulching straw (S), conifer compost (CC), and conifer bark (CB) as well as grass landing with grass (G), clover (Cl), and fescue (F) in a silty-clay soil (Macon), and by incorporating vine shoot (VS) and single and double doses of farmyard manure (FM) and mushroom manure (MM) in a calcareous sandy soil (Chinon). Soil physicochemical and microbial characteristics were assessed at each site at two depths by sampling at 0-5 and 5-20 cm for the Macon site and 0 10 and 10-20 cm for the Chinon site. Changes in the quantity of soil organic matter (SOM), through an increase in C(org) and N(org) contents, and in its quality, through modifications in the C/N and humic acid/fulvic acid ratios, were essentially recorded at the surface layer of treated plots with differential magnitudes according to the inputs and soil type. Quantitative modifications in microbial communities were assessed by means of C-biomass measurements and resulted in an increase in microbial densities fitted with the increase of C(org) and N(org) contents. However, the deduced C incorporation in microbial biomass was negatively correlated with the C/N ratio, demonstrating a strong influence of the type of organic management on the rate of microbial processes. Qualitative modifications in microbial communities were evaluated by the characterization of the genetic structure of bacterial and fungal communities from DNA directly extracted from the soil, using bacterial and fungal automated ribosomal intergenic spacer analysis. Organic amendments led to changes in the bacterial and fungal communities of both sites. However, the magnitude and the specificity of these changes were different between sites, organic amendments, and microorganisms targeted, revealing that the impact of organic management is dependent on the soil and organic input types as well as on the particular ecology of microorganisms. A co-inertia analysis was performed to specify the role of the quantity and quality of SOM on the modifications of the genetic structure. A significant costructure was only observed for Macon plots at 0-5 cm between the bacterial genetic structure and the SOM characteristics, demonstrating the influence of the relative amount of the different humic substances (humic and fulvic acids) on microbial composition. PMID- 17401598 TI - Embolization for massive urethral hemorrhage. AB - Priapism is the term given to a prolonged, painful penile erection, unaccompanied by sexual desire. Most cases of priapism occur as a result of engorgement of the cavernous body of the penis due to intrinsic or extrinsic venous outflow obstruction, which is termed veno-occlusive priapism. However, in a small number of patients, the cause is related to uncontrolled arterial inflow due to trauma and resultant laceration of cavernosal arteries. This form of disease has been termed high-flow priapism and usually occurs in association with genitoperineal trauma. Most incidents of high-flow arteriogenic priapism result from genitoperineal trauma, such as that caused by sliding on a wooden bar or falling astride a ladder. We experienced a patient with a high-flow priapism due to urethral trauma caused by a misplaced Foley catheter. The treatment consisted of superselective arterial embolization of the bilateral internal pudendal arteries using metallic microcoils, which resulted in the subsequent detumescence of the penis with no complications. This case showed that bilateral arteriocavernosal fistulae can be successfully treated by superselective arterial embolization. PMID- 17401599 TI - Seminoma in undescended testis. AB - A testicular carcinoma in intraabdominal undescended testis can mimic other mesenteric or retroperitoneal mass. The identification of pampiniform plexus draining the mass can play an important role in diagnosis of testicular carcinoma in undescended testis. We report a case of seminoma in intraabdominal undescended testis, draining into pampiniform plexus and thrombus within it. PMID- 17401600 TI - RFA of renal cell carcinoma in a solitary kidney. AB - The incidence of renal cell carcinoma (RCC) is increasing worldwide and diagnosis is made in an earlier stage due to increasing use of CT and ultrasound. Patients with a history of RCC have a high risk to develop another RCC within their remaining kidney. Especially, patients after tumor nephrectomy have to undergo follow-up examinations regularly allowing an early detection of new RCCs. Especially in patients with solitary kidney gold standard therapy (nephron sparing surgery) can often account for consecutive renal failure with the need for hemodialysis. Percutaneous radiofrequency ablation (RFA) gained worldwide acceptance for the treatment of liver tumors in patients unable to undergo surgery. Furthermore, during the past few years, there is an increasing amount of publications dealing with the effectiveness of minimally invasive therapies like RFA in patients with renal masses. However, in the subgroup of patients with solitary kidney suffering from RCC, there are only case reports regarding safety and midterm outcome available so far. Therefore, the aim of this article is to briefly describe the basic technical principles of RFA and then focus on indications, technique, safety and the midterm outcome after treatment in terms of renal function and relapse. PMID- 17401601 TI - Pancreatic transplants: secretin-stimulated MR pancreatography. AB - BACKGROUND: Our study was aimed to evaluate the functional status of pancreatic transplants using dynamic MR pancreatography after secretin stimulation. METHODS: Thirteen asymptomatic patients previously submitted to isolated pancreas (n = 6) or combined kidney-pancreas (n = 7) transplantation, with enteric-portal pancreatic drainage, underwent MR examination at 1.5 T using a phased-array coil. After the acquisition of axial and coronal T1- and T2-weighted sequences, dynamic MR pancreatography was performed using a coronal breath-hold, thick-slab (40-60 mm), single-shot T2-weighted fast spin-echo sequence. After the intravenous administration of secretin (Secrelux, Sanochemia; 1 cU/kg body/weight), a single slice image acquisition was repeated every 30 s up to 15 min. We estimated the calibre changes of the pancreatic ductal system and the filling of the donor's duodenum on the basis of pancreatic secretion after secretin stimulation, also evaluated by using a mean signal intensity/time histogram in a chosen region of interest including the transplanted pancreas and the connected small bowel. RESULTS: All patients well tolerated the examination, and no side effects were reported after secretin administration. In 12/13 cases, a significant increase (more than 1 mm) in the diameter of the mean pancreatic duct was observed after secretin stimulation; in all patients, a noticeable filling of the duodenal graft was demonstrated during dynamic MR pancreatography on both qualitative and quantitative analyses. CONCLUSIONS: Dynamic MR imaging after secretin administration allows non-invasive evaluation of exocrine function of the pancreatic transplants and could be used to differentiate patients with graft rejection from those with normal graft function. PMID- 17401602 TI - Reversal of nicotine-induced alveolar lipofibroblast-to-myofibroblast transdifferentiation by stimulants of parathyroid hormone-related protein signaling. AB - Nicotine exposure disrupts the parathyroid hormone-related protein (PTHrP)-driven alveolar epithelial-mesenchymal paracrine-signaling pathway, resulting in the transdifferentiation of pulmonary lipofibroblasts (LIFs) to myofibroblasts (MYFs), which seems to be central to altered pulmonary development and function in infants born to mothers who smoke during pregnancy. Modulation of PTHrP-driven signaling can almost completely prevent nicotine-induced LIF-to-MYF transdifferentiation. However, once this process has occurred, whether it can be reversed is not known. Our objective was to determine if nicotine-induced LIF-to MYF transdifferentiation could be reversed by specifically targeting the PTHrP mediated alveolar epithelial-mesenchymal paracrine signaling. WI38 cells, a human embryonic pulmonary fibroblast cell line, were initially treated with nicotine for 7 days and LIF-to-MYF transdifferentiation was confirmed by determining the downregulation of the key lipogenic marker, peroxisome proliferator-activated receptor gamma (PPARgamma) and upregulation of the key myogenic marker, alpha smooth muscle actin (alphaSMA). Because downregulation of the PPARgamma signaling pathway is the key determinant of LIF-to-MYF transdifferentiation, cells were treated with three agonists of this pathway, PTHrP, dibutryl cAMP (DBcAMP), or rosiglitazone (RGZ) for 7 days, and the expression of the PTHrP receptor, PPARgamma, alphaSMA, and calponin was determined by Western analysis and immunohistochemistry. Simultaneously, fibroblast function was characterized by measuring their capacity to take up triglycerides. Nicotine-induced LIF-to-MYF transdifferentiation was almost completely reversed by treatment with RGZ, PTHrP, or DBcAMP, as determined by protein and functional assays. Using a specific molecular approach and targeting specific molecular intermediates in the PTHrP signaling pathway, to our knowledge, this for the first time, demonstrates the reversibility of nicotine-induced LIF-to-MYF transdifferentiation, suggesting not only the possibility of prevention but also the potential for reversal of nicotine-induced lung injury. PMID- 17401603 TI - Endoluminal GERD treatments: critical appraisal of current literature with evidence-based medicine instruments. AB - BACKGROUND: The literature of endoluminal treatment of gastroesophageal reflux disease (GERD) widely varies in the level of evidence presented for analysis. Therefore there is a need for a comprehensive evidence-based medicine (EBM) analysis of the current literature evidence of the three FDA-approved modalities used for endoluminal treatment of GERD. SEARCH STRATEGY: In January 2007, the MEDLINE database was searched for randomized controlled trials (RCTs), and controlled clinical trials of currently available endoluminal treatment of GERD. Database searches combined the specific endoluminal device keywords with the condition-specific keyword (e.g., GERD). DATA COLLECTION AND ANALYSIS: All relevant studies have been categorized according to the evidence they provide according to the guidelines for Levels of Evidence and Grades of Recommendation supplied by the Oxford Centre for Evidence-Based Medicine. MAIN RESULTS AND AUTHORS' CONCLUSION: Sixteen studies met the inclusion criteria, representing 787 patients. The methodological quality of most of the included studies was average; four studies were grade 1b (individual randomized trial), 10 were grade 2b (individual cohort study), and two were grade 3b (individual case-control study) There is grade 1b and 2b evidence demonstrating the EndoCinch plication is effective in reducing GERD symptoms at short-term follow up. However, in the majority of the studies analyzed, the procedure does not significantly reduce the acid exposure in the distal esophagus. The majority of the studies with long-term outcome showed disappointing outcomes, probably due to suture loss in the majority of patients. There is grade 1b and 2b evidence demonstrating that the Stretta procedure is effective in reducing GERD symptoms at short- and mid-term follow up. However, in the majority of the studies analyzed, the procedure did not reduce significantly the acid exposure in the distal esophagus. There is grade 1b and 2b evidence demonstrating that full-thickness plication is effective in reducing GERD symptoms, and acid exposure in the distal esophagus. PMID- 17401604 TI - Similarity of traveling-wave delays in the hearing organs of humans and other tetrapods. AB - Transduction of sound in mammalian ears is mediated by basilar-membrane waves exhibiting delays that increase systematically with distance from the cochlear base. Most contemporary accounts of such "traveling-wave" delays in humans have ignored postmortem basilar-membrane measurements in favor of indirect in vivo estimates derived from brainstem-evoked responses, compound action potentials, and otoacoustic emissions. Here, we show that those indirect delay estimates are either flawed or inadequately calibrated. In particular, we argue against assertions based on indirect estimates that basilar-membrane delays are much longer in humans than in experimental animals. We also estimate in vivo basilar membrane delays in humans by correcting postmortem measurements in humans according to the effects of death on basilar-membrane vibrations in other mammalian species. The estimated in vivo basilar-membrane delays in humans are similar to delays in the hearing organs of other tetrapods, including those in which basilar membranes do not sustain traveling waves or that lack basilar membranes altogether. PMID- 17401605 TI - Anticoagulation control in the peri-hospitalization period. AB - Oral anticoagulants (OAC) are effective and safe if the international normalized ratio (INR) is maintained within a narrow therapeutic range. Hospitalization is independently associated with poor anticoagulation control. The objective of this study is to describe how anticoagulation control changes in the peri hospitalization period. This study is a retrospective cohort study using population-based administrative databases. INR results were retrieved from a population-based laboratory database. INR levels between laboratory measures were estimated using linear interpolation. Auto-regressive, integrated, moving average (ARIMA) time-series modeling was used to determine how anticoagulation control changed in the peri-hospitalization period. The study included 5,380 elderly patients in Eastern Ontario between 1 September 1999 and 1 September 2000 taking OACs. Results showed that 951 (17.7%) were hospitalized during their OAC therapy [thrombotic, n = 52 (1.0%); hemorrhagic, n = 140 (2.6%); other hospitalization types, n = 759 (14.1%)]. All measures of anticoagulation control changed significantly in the peri-hospitalization period. Before hemorrhagic admissions, mean INR and proportion with INR > 5 increased significantly (daily increase 0.024, P = .03 and 0.2%, P = .01). Following other hospitalization types, the proportion of patients with INR < 1.5 was significantly increased (daily increase 0.19%, P = .02). Patients admitted to the hospital for a variety of indications have significantly worse anticoagulation control in the peri-hospitalization period. Anticoagulated patients discharged after medical hospitalizations could be targeted for improved anticoagulation control. PMID- 17401606 TI - Long-acting medications for the hyperkinetic disorders. A note on cost effectiveness. AB - New long-acting medications for attention-deficit/ hyperactivity disorder (ADHD) have become available, which combine certain advantages over conventional short acting drugs with higher acquisition costs. Choices between these drugs should thus be driven by their clinical profiles and by an acceptable balance of increased costs and additional benefits. Accordingly, the notion of relative cost effectiveness should be central to recommendations about the use of these drugs in practice. A recent technology assessment on behalf of the National Institute for Health and Clinical Excellence (NICE) did not identify differences between compounds in terms of clinical efficacy and described drug cost as the major driver of cost-effectiveness. The underlying economic model was restricted to a cost-utility analysis that used only a fraction of the available clinical evidence base and did not address the distinction between efficacy and effectiveness. Cost-effectiveness evaluations including the potential impact of improved treatment compliance indicate a relatively more attractive cost effectiveness of long-acting medications than suggested by the NICE assessment. These evaluations provide health economic support to treatment recommendations recently published by the European Network for Hyperkinetic Disorders. Limitations of currently available economic evaluations include their short time horizon, and future research should assess treatment effects on long-term sequelae associated with ADHD. PMID- 17401607 TI - Stability and change of IQ scores in preschool children diagnosed with autistic spectrum disorder. AB - AIM: To investigate cognitive development in preschool-age children diagnosed with Autistic Spectrum Disorder (ASD; N = 39) compared with that of children diagnosed with mental retardation (MR; N = 14) and normally developing children (NC; N = 36). METHOD: In a prospective longitudinal study, cognitive development was tested at age 24 months (T1; SD = 6 months) and 43 months (T2; SD = 5). RESULTS: Group IQ scores were stable between T1 and T2 as evidenced by high correlations (r = .81, P < .01) and consistency of average group scores. At the same time however, about a third of children with ASD showed an increase of cognitive scores of 15 points or more. This increase of IQ was correlated with lower scores at the early screening of autistic traits (ESAT) at T1, higher IQ level at T2 and higher expressive language skills at T2. Intensity of treatment was not related to IQ increase. CONCLUSIONS: High correlations between cognitive scores in preschool children with ASD suggest that measurements of cognitive function are valid at this age. We found indications of both stability and change of IQ scores. Findings suggest that some children with ASD show catch-up intellectual development. To the best of our knowledge, this increase in IQ scores cannot be attributed to treatment effects. PMID- 17401608 TI - Selective difficulty in recognising facial expressions of emotion in boys with ADHD. General performance impairments or specific problems in social cognition? AB - Research on emotion understanding in ADHD shows inconsistent results. This study uses control methods to investigate two questions about recognition and understanding of emotional expressions in 36 five- to eleven-year-old boys with ADHD: [1] Do they find this task more difficult than judging non-emotional information from faces, thus suggesting a specific social-cognitive impairment? [2] Are their judgements about faces impaired by general limitations on task performance, such as impulsive responding? In Part 1, 19 boys with ADHD and 19 age-matched typically developing boys matched facial expressions of emotion to situations, and did a control non-emotional face-processing task. Boys with ADHD performed more poorly than age-matches on both tasks, but found the emotion task harder than the non-emotion task. In Part 2, 17 boys with ADHD and 13 five-to six year-old typically developing boys performed the same tasks, but with an 'inhibitory scaffolding' procedure to prevent impulsive responding. Boys with ADHD performed as well as the younger controls on the non-emotional task, but still showed impairments in the emotion task. Boys with ADHD may show poorer task performance because of general cognitive factors, but also showed selective problems in matching facial emotions to situations. PMID- 17401609 TI - Evaluation of family drawings of physically and sexually abused children. AB - OBJECTIVE: The aim of this study is to analyse the family drawings of two groups of physically and/or sexually abused children as compared to the drawings of non abused children of a matched control group. METHODS: The drawings by 12 physically abused, 12 sexually abused and 12 non-abused children, all aged between 5 years-old and 10 years-old, were assessed and compared. Family drawings were analysed using a specific Screening Inventory (FDI-Family Drawing Inventory). This Inventory takes into consideration such qualitative and quantitative variables as the quality of drawing, the children's perception of their family members and their own perception of themselves within the family system. RESULTS: The results have shown significant differences between the abused minors and the control group. Abused children are more likely to draw distorted bodies, the human figure is usually represented devoid of details, their drawings generally show clear signals of trauma and the majority of the abused children are likely to exclude their primary caregiver from the drawings. CONCLUSIONS: The "drawings of the family" of physically and/or sexually abused children significantly evidence a greater emotional distress then the drawings of the non-abused children of the matched control group. PMID- 17401610 TI - Childbirth complications affect young infants' behavior. AB - BACKGROUND: The process of childbirth and its complications have been related to the newborn's condition and to development at later ages. In this study, we examine how mode of delivery and delivery complications are related to the behavior and cortisol reactivity of infants during the first 2 months. METHODS: Delivery factors (i.e. mode, duration, fetal heart function, and 5-min Apgar score) were determined in 116 healthy, term, firstborn infants. The infants' behavioral and cortisol reactivity to stressors were assessed during a physical examination at 11 days and an inoculation at 2 months. Daily behavior at 6 weeks was followed by means of a four-day parental diary. RESULTS: Indicators of a more stressful delivery were linked to more crying/fussing in the infant and to more difficulties in regulating the infants' behavior (i.e., more unsoothability and longer crying bouts). Specific delivery complications rather than mode of delivery were linked to infants' behavior. No significant associations however, were found between childbirth and the infants' cortisol reactivity to a physical examination and an inoculation. CONCLUSIONS: Childbirth complications, even when mild, can apparently affect early infant behavior. These effects extend over different types of situations, and last at least until the age of 2 months, and should be taken into account in studies on infant behavior. Earlier findings of relations between childbirth complications and infant cortisol reactivity could not be replicated in this study, possibly due to the confounding effect of parity, and to the important intra-individual variability present in the infants' cortisol. PMID- 17401611 TI - Changes in social competence in young children treated because of conduct problems as viewed by multiple informants. AB - In the present study changes in social competence were examined in a clinic sample of 127 children aged 4-8. The children were recruited to a controlled treatment study because of conduct problems at home and were randomised to the Incredible Years parent training (PT), combined PT and child therapy (CT) or a waiting-list control-group. Assessments were conducted pre- and post-treatment and at a one-year follow-up by multiple informants (mother, father, teacher and child). Parent training combined with child treatment showed most improvement in child social competence based on mother, father and child reports, however, father reports showed positive results for children treated with PT only. Treated mothers and fathers showed a decrease in correlations in their reports of social competence in the child after treatment as compared to parents in the waiting list condition. No generalisation effects to peer-relationships in day care/school were found, neither on teacher or child reports. A broad perspective using multiple informants from different settings is needed when effects of treatment of young children with conduct problems are evaluated and should include various aspects of social competence. PMID- 17401612 TI - ADHD and comorbid disorders in relation to family environment and symptom severity. AB - BACKGROUND: To examine the comorbidity of ADHD in association with family environment and the severity of ADHD. METHOD: A screening for ADHD symptoms was conducted among adolescents in the Northern Finland 1986 Birth Cohort (N = 6622). A sample of those adolescents (n = 457), aged 16-18 years, with and without ADHD symptoms was assessed with a diagnostic interview (Kiddie-SADS-PL) and ADHD and comorbid disorders were studied in association with the family characteristics and the number of ADHD symptoms. RESULTS: Adolescents with ADHD had more commonly conduct disorder (P < 0.001), oppositional defiant disorder (P < 0.001), substance abuse (P < 0.001) and mild depression (P < 0.001) than adolescents without ADHD. Adolescents with ADHD and comorbid disorders had more ADHD symptoms (P < 0.001) than those with ADHD alone. Compared to adolescents with ADHD alone those with ADHD and comorbidity lived significantly more commonly in non-intact families, in low-income families, with mothers who were dissatisfied with life and with parents who showed little interest in their adolescents' activities. CONCLUSIONS: Adolescents who develop externalizing disorders comorbid to ADHD seem to suffer from a severe form of ADHD and live in family environments that may not provide sufficient support for optimal development of an adolescent with ADHD. PMID- 17401613 TI - Physiological correlates of anxiety in children with gender identity disorder. AB - BACKGROUND: Internalizing psychopathology, anxiety in particular, is assumed to contribute to the development of gender Identity disorder (GID). Until now, anxiety has only been reported in studies using parent-report questionnaires; physiological correlates of anxiety have not been studied. In this study we assessed anxiety and stress in children with GID by measuring their cortisol, heart rate (HR) and skin conductance levels (SCL) and asking them to repeat their moods and experience of control. METHODS: By using an established psychological challenge, involving provocation and frustration, we investigated whether children with GID as compared to healthy controls react in a more anxious way under these experimental circumstances. We assessed anxiety levels in 25 children with GID and 25 matched controls by measuring cortisol, HR and skin conductance and examined whether a pattern of increased physiological activity corresponded with the feeling of being less in control and having more intense negative emotions. RESULTS: The results showed that children with GID had more negative emotions and a tonically elevated SCL. There were no differences between the groups in cortisol and HR. CONCLUSION: This is the first study that shows that children with GID have a more anxious nature as compared to their normal counterparts. PMID- 17401614 TI - Autism spectrum disorder and Klinefelter syndrome. AB - BACKGROUND: Autism is a severe handicapping disorder of early childhood characterized by a distinct pattern of social and communication impairment with rigid ritualistic interests. In about 10-25% of cases, it is associated with known medical conditions. Population-based studies of autism have found that Klinefelter's syndrome (KS), a common chromosome abnormality, is sometimes associated with autism. However, few detailed case descriptions of patients with KS and autism have not been published. CASE REPORT: In this paper, we describe the occurrence of autistic features in two cases of Klinefelter syndrome, one with the typical XXY karyotype and the other with the XXYY variant. CONCLUSION: Autistic features may be more common in persons with Klinefelter syndrome than generally believed. We propose that all patients with KS should be screened for the presence of autism. PMID- 17401615 TI - The evolutionary dynamics of canid and mongoose rabies virus in Southern Africa. AB - Two variants of rabies virus (RABV) currently circulate in southern Africa: canid RABV, mainly associated with dogs, jackals, and bat-eared foxes, and mongoose RABV. To investigate the evolutionary dynamics of these variants, we performed coalescent-based analyses of the G-L inter-genic region, allowing for rate variation among viral lineages through the use of a relaxed molecular clock. This revealed that mongoose RABV is evolving more slowly than canid RABV, with mean evolutionary rates of 0.826 and 1.676 x 10(-3) nucleotide substitutions per site, per year, respectively. Additionally, mongoose RABV exhibits older genetic diversity than canid RABV, with common ancestors dating to 73 and 30 years, respectively, and while mongoose RABV has experienced exponential population growth over its evolutionary history in Africa, populations of canid RABV have maintained a constant size. Hence, despite circulating in the same geographic region, these two variants of RABV exhibit striking differences in evolutionary dynamics which are likely to reflect differences in their underlying ecology. PMID- 17401616 TI - Intraocular use of hydrogel tissue adhesive in rabbit eyes. AB - PURPOSE: To evaluate the safety and efficacy of newly developed hydrogel glue to treat rhegmatogenous retinal detachments in in vitro and in vivo studies. METHODS: In the in vitro study, the solid hydrogel glue was soaked in a balanced salt solution (BSS). The pH was measured periodically, and the dissolution time was recorded. In the in vivo study in six albino rabbits, 0.1 ml of hydrogel glue was injected into the right vitreous cavity, and 0.1 ml BSS was injected into the left vitreous, as the control. Clinical, electroretinography (ERG) and histological examinations were performed. Retinal detachment with a retinal hole was created in 12 albino rabbits after vitrectomy. After fluid-air exchange, the hydrogel glue was applied to the hole in nine rabbits; three other rabbits were used as controls. Clinical and histological examinations were performed. RESULTS: The pH ranged from 6.65 to 8.14. The glue remained solid in BSS for 7 weeks. The glue injection did not induce inflammation. There was no significant difference between the study and control eyes in the ERG amplitude or the implicit times of the a and b waves. No significant histological abnormality was detected. The retina was reattached with glue in three of nine eyes. The histological examination showed glue under the retina. CONCLUSIONS: Hydrogel glue, which seemed to be minimally toxic to the eye, can be used to patch retinal breaks. However, methods to mix a small amount of each solution completely and a more effective intraocular delivery system for the glue are needed. PMID- 17401617 TI - Exclusion of transforming growth factor-beta1 as a candidate gene for myopia in the Japanese. AB - PURPOSE: To determine whether single-nucleotide polymorphisms (SNPs) within the transforming growth factor-beta1 (TGF-beta1) gene are associated with high myopia in Japanese. Previous studies have indicated that the gene expression products, regulators of the TGF-beta1 gene, are involved in high myopia. METHODS: Genomic DNA samples were obtained from 330 Japanese patients with high myopia and 330 Japanese controls without high myopia who were chosen at random. SNPs were genotyped by the TaqMan system, using primer extension and polymerase chain reaction amplification. RESULTS: Ten SNPs were identified in the high-myopia patients and controls, with four of the ten SNPs having nonsynonymous changes. However, no statistical differences in the SNPs were detected between the high myopia cases and the controls. CONCLUSIONS: Sequence variants of the TGF-beta1 gene were not associated significantly with high myopia, and further studies are needed to identify which genes are responsible for high myopia. PMID- 17401618 TI - Soluble IL-6 receptor in vitreous fluid of patients with proliferative diabetic retinopathy. AB - PURPOSE: To identify the biological reaction of soluble interleukin-6 receptor (sIL-6R) in the vitreous of patients with proliferative diabetic retinopathy (PDR). METHODS: The subjects were 45 patients (45 eyes) with vitreoretinal diseases. The patients were divided into three groups: the PDR group comprised 28 patients (28 eyes) with PDR; the pre-proliferative diabetic retinopathy (PPDR) group comprised seven patients (seven eyes) with PPDR combined with diabetic macular edema; and the nondiabetic group comprised ten patients (ten eyes) with idiopathic macular hole or idiopathic epiretinal membrane. Vitreous samples were obtained at vitrectomy. sIL-6R, vascular endothelial growth factor (VEGF), and protein concentration in vitreous samples were determined by enzyme-linked immunosorbent assay (ELISA). sIL-6R levels in serum were also determined by ELISA in nine of the 28 patients with PDR and in six healthy volunteers as controls. RESULTS: In vitreous fluid, the levels of sIL-6R in the PDR group, PPDR group, and nondiabetic group were 612.7 +/- 233.8 (mean +/- SD), 746.3 +/- 523.1, and 215.4 +/- 98.3 pg/ml, respectively. Vitreous levels of sIL-6R in the PDR and PPDR groups were significantly higher than those in the nondiabetic group (PDR group, P < 0.0001; PPDR group, P < 0.01). In serum, the levels of sIL-6R were 39.38 +/- 9.43 ng/ml in the PDR group and 22.84 +/- 5.32 ng/ml in the control group. sIL-6R levels in serum in the PDR group were significantly higher than those in the control group (P < 0.01). A partial correlation analysis showed a significant correlation between the levels of sL-6R and VEGF in the vitreous in the PDR group (r = 0.34, P < 0.05). CONCLUSIONS: We conclude that the level of sIL-6R in vitreous fluid can be considered as a biomarker of PDR. PMID- 17401619 TI - Axial length measurement using partial coherence interferometry in myopic children: repeatability of the measurement and comparison with refractive components. AB - PURPOSE: To evaluate the test-retest repeatability of axial length (AL) measurements made with the IOLMaster in children with a wide range of myopic refractive errors, and to clarify the relationship between the AL and other refractive components. METHODS: The subjects were 95 children (mean age +/- SD, 10.8 +/- 1.3 years; range, 7-13 years; mean refractive error; -4.37 +/- 1.43 D; range, -1.50 to -8.19 D) who participated in a myopia-control trial. The AL of the right eye was measured three times using an IOLMaster, and the mean value was regarded as the representative measurement. After 5 min, the measurement was performed again, and the repeatability was evaluated by analyzing the distribution of differences between the two measurements. The relationships between the AL and age, cycloplegic autorefraction (RE), and corneal radius of curvature (CR) were also examined. RESULTS: The repeatability of the IOLMaster measurements was +/-0.05 mm (corresponding to a refractive error of +/-0.12 D) and was not affected by age or RE. AL was negatively correlated with RE (RE = 0.68 x AL + 12.74, r = -0.37) and positively correlated with CR (CR = 0.21 x AL + 2.53, r = 0.69). The highest correlation was found between the ratio of AL to CR and RE (AL/CR = -0.04 x RE + 3.08, r = -0.76). No association was observed between age and AL, nor between CR and RE. CONCLUSIONS: The IOLMaster provides high repeatability in AL measurement in myopic children. The associations between AL and each refractive component found in this study were consistent with those in previous studies that used the ultrasound A-mode method, except for slightly higher AL/CR ratios in our study. PMID- 17401620 TI - Cycloplegic effect of 0.5% tropicamide and 0.5% phenylephrine mixed eye drops: objective assessment in Japanese schoolchildren with myopia. AB - PURPOSE: To evaluate the cycloplegic effect of mixed eye drops containing 0.5% tropicamide and 0.5% phenylephrine in myopic children, and to determine whether their efficacy was associated with their clinical characteristics. METHODS: Eighty-one myopic children (age, mean +/- SD, 11.0 +/- 1.5 years; mean spherical equivalent refractive error, -4.27 +/- 1.41 D; range, -1.57 to -8.66 D) were recruited. One drop of Mydrin-P was administered to each eye twice, with an interval of 5 min between. Twenty-five minutes after the second drop, accommodative responses were measured with an open-view autorefractometer, while the subject was encouraged to accommodate by binocularly looking at a Maltese cross located at a distance of 33 cm. The difference between the refractive reading and that obtained with a Maltese cross at 500 cm was regarded as residual accommodation (RA). The repeatability of this measurement was also evaluated. RESULTS: The mean RA was 0.21 +/- 0.29 D (range, -0.31 to 0.99 D). There was no association in RA between the right and left eyes, between RA and age, or between RA and sex, but RA was weakly correlated with refractive error (r = 0.274, P = 0.019). The intersubject difference found in RA can be explained mostly by the extent of repeatability (+/-0.71 D). CONCLUSION: The insignificant magnitude of RA indicated that the mixed eye drop is an acceptable and useful cycloplegic agent in Japanese schoolchildren with a wide range of myopic refractive errors. PMID- 17401621 TI - Stability of the central visual field after modern trabeculectomy techniques in eyes with advanced glaucoma. AB - PURPOSE: To review the surgical results and complications of trabeculectomy techniques in patients with advanced glaucoma and threatened fixation. METHODS: Trabeculectomy had been carried out on 49 advanced glaucoma patients (49 eyes) using mitomycin C and postoperative laser suture lysis. The clinical records prior to and 2 months after surgery were reviewed, and the long-term surgical outcomes were determined. RESULTS: Two months after surgery there were no eyes with fixation loss. Intraocular pressure (IOP) levels were reduced from 22.8 +/- 6.0 to 11.7 +/- 4.7 mmHg. Kaplan-Meier survival analysis showed that the success rate in achieving IOPs of 15 mmHg or lower 5 years after surgery was 70%. The chance of visual acuity remaining within two lines of the preoperative level was 75%. In 29 of the 49 eyes, visual acuities remained at their preoperative level at the time of the final visit, but had decreased to less than 0.1 in three eyes (cataract progression, n = 2; fixation loss, n = 1). CONCLUSION: The results suggest that laser suture lysis and stepwise management of IOP levels, which are performed as part of the modern postoperative management of trabeculectomy, decrease the frequency of fixation loss during the early postsurgical phase. PMID- 17401622 TI - Evaluation of characteristic ocular signs and systemic investigations in ocular sarcoidosis patients. AB - PURPOSE: To evaluate the diagnostic values of ocular signs and systemic investigations in ocular sarcoidosis, in a retrospective case-control study. METHODS: Subjects were 67 consecutive uveitis patients with biopsy-proven sarcoidosis and 111 control patients with other clinical uveitis entities. The predictive values analyzed were sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The five ocular signs for ocular sarcoidosis are (1) mutton fat keratic precipitates and iris nodules; (2) nodules at the trabecular meshwork and tent-shaped peripheral anterior synechia; (3) snowball vitreous opacities; (4) nodular periphlebitis, and (5) multiple chorioretinal lesions (active or atrophic) in the peripheral fundus. In addition, the results of the following five systemic investigations were considered: (1) negative tuberculin skin test; (2) elevated serum angiotensin-converting enzyme; (3) elevated serum lysozyme; (4) elevated serum gamma-globulin; and (5) bilateral hilar lymphadenopathy on chest X-ray. RESULTS: The incidence of all ocular signs and positive results for the systemic investigations were significantly higher in sarcoidosis patients than in controls (P < 0.001). The presence of two or three of the five ocular signs were indicative of a positive finding in the diagnostic parameters. The presence of two positive results among the five systemic investigations showed values higher than 0.800 for all diagnostic parameters. CONCLUSIONS: Combinations of the specified ocular signs and the results of systemic investigations can be used for the diagnosis of ocular sarcoidosis. PMID- 17401623 TI - Histological study of choroidal malignant melanoma treated by carbon ion radiotherapy. AB - PURPOSE: To report, we believe for the first time, a histological study of choroidal malignant melanoma treated by carbon ion beam radiotherapy. METHODS: A 75-year-old Japanese man was diagnosed as having a choroidal melanoma after undergoing magnetic resonance imaging (MRI). Positron emission tomography (PET) revealed a hot spot in the same location as the intraocular mass seen in MRI. Carbon ion radiotherapy was performed with a total dose of 77 Gy, and the hot spot seen by PET disappeared completely. At 15 months after carbon ion therapy, the eye had to be enucleated because of uncontrollable ocular hypertension. It was examined histologically in serial sections. RESULTS: A large tumor mass (15 x 12 mm) with high pigmentation was found in the vitreous space. Almost all tumor cells showed necrosis in every section. A small number of intact tumor cells were present at the periphery. The overlying retina did not show any necrosis, but showed mild to moderate gliosis. No intraretinal hemorrhage, lipid deposit, or protein exudate was apparent. CONCLUSIONS: Almost all tumor cells showed necrosis after radiotherapy with a carbon ion beam. However, the effect on the adjacent tissues was determined as minimal in histological analysis. PMID- 17401624 TI - Eye manifestations in patients with perinuclear antineutrophil cytoplasmic antibody-associated vasculitis: case series and literature review. AB - PURPOSE: To report and summarize eye manifestations of patients with perinuclear pattern antineutrophil cytoplasmic antibody [pANCA, myeloperoxidase (MPO)-ANCA] associated vasculitis. METHODS: The medical records of four consecutive patients with pANCA (MPO-ANCA) vasculitis who showed eye manifestations were retrospectively reviewed. In addition, the medical literature databases, PubMed and Japana Centra Revuo Medicina for Japanese literature, were searched for pANCA vasculitis patients with eye manifestations. RESULTS: Three of the four patients treated at the Okayama University Hospital showed unilateral or bilateral scleritis. In the literature review, eight of the 27 patients showed ocular surface manifestations such as scleritis and peripheral keratitis. Other frequent eye presentations were posterior segment manifestations such as central or branch retinal vein occlusion, optic neuropathy, and acute posterior multifocal placoid pigment epitheliopathy (APMPPE). Systemically, the most frequent manifestations were glomerulonephritis in the present patients as well as in the patients reported in the past studies. CONCLUSIONS: Ocular surface manifestations and posterior segment manifestations were major eye presentations in patients with pANCA-associated vasculitis. ANCA testing including both pANCA and cytoplasmic pattern antineutrophil cytoplasmic antibody would help establish a systemic diagnosis in patients with eye manifestations such as scleritis, retinal vein occlusion, optic neuropathy, or APMPPE. PMID- 17401625 TI - Simultaneous conjunctival, uveal, and orbital involvement as the initial sign of acute lymphoblastic leukemia. AB - BACKGROUND: A case of acute lymphoblastic leukemia with an initial presentation of unilateral simultaneous conjunctival, uveal, and orbital infiltration is reported. CASE: A 9-year-old girl presented with mild proptosis and chemosis and total exudative retinal detachment in the right eye. OBSERVATIONS: B-mode ultrasonography showed diffuse uveal thickening and a retrobulbar mass. Magnetic resonance imaging scans demonstrated a well-circumscribed peribulbar orbital mass. Incisional biopsy from the conjunctiva and bone marrow sampling established the diagnosis of acute lymphoblastic leukemia L1 type. CONCLUSION: Blast-cell infiltration of three distinct ocular structures simultaneously in the absence of any peripheral signs is an extremely rare initial manifestation of acute lymphoblastic leukemia. PMID- 17401627 TI - Traumatic wound dehiscence after penetrating keratoplasty. PMID- 17401626 TI - A case of incontinentia pigmenti in Japan and its genetic examination. AB - PURPOSE: Incontinentia pigmenti (IP) is a rare X-linked dominant genodermatosis with approximately one-third of cases with associated ocular complications. Deletion of exons 4 to 10 of the nuclear factor kappaB essential modulator (NEMO) gene accounts for the majority of new mutations. The disease is more commonly found among Caucasians. We studied a case of an IP patient in Japan, and the genomic rearrangements. METHODS: An 11-month-old female infant exhibited the skin lesions of IP. Ocular findings were total retinal detachment with a retrolental fibrovascular mass in the right eye, and patchy retinal avascular zones and neovascularization in the left eye. The genomic rearrangement of NEMO was investigated by a polymerase chain reaction (PCR)-based diagnostic test. A skewed X-inactivation assay was also performed using the human androgen receptor gene as a genetic marker. RESULTS: Deletion of exons 4 to 10 in NEMO was detected in the proband and in other female members of her family. A complete skewing of the X inactivation pattern causing IP was observed, indicating cells having no protection against apoptosis in response to tumor necrosis factor as the pathogenicity of the disease. CONCLUSION: This is the first case report of a Japanese female phenotype demonstrating the common genomic rearrangement in the NEMO gene. PMID- 17401628 TI - B cell clonality of primary central nervous system and primary intraocular lymphomas. PMID- 17401629 TI - Primary conjunctival follicular lymphoma treated with the anti-CD20 antibody rituximab and low-dose involved-field radiotherapy. PMID- 17401630 TI - Giant pituitary adenoma manifesting as homonymous hemianopia. PMID- 17401631 TI - Case report of multiple sclerosis in which visual function was evaluated using multifocal visual evoked potentials. PMID- 17401633 TI - Prognostic factors for progression of visual field damage in patients with normal tension glaucoma. PMID- 17401634 TI - Allassogonoporus dromiciops n. sp. (Digenea: Allassogonoporidae) from Dromiciops gliroides (Marsupialia: Microbiotheriidae) in Patagonia, Argentina. AB - A new species of Allassogonoporus Olivier, 1938, from a naturally infected specimen of Dromiciops gliroides, is described. Adults of A. dromiciops n. sp. were recovered from the small intestine. This new species can be distinguished from all other species of the genus by having: the largest body and eggs, a lobed testes and ovary, a Y-shaped excretory vesicle and vitelline follicles beginning at the level of the posterior margin of the oral sucker. This is the first record of a species of Allassogonoporus in South America and the first record of a digenean parasitising D. gliroides. PMID- 17401635 TI - Molecular characterization of two Arabidopsis thaliana glycosyltransferase mutants, rra1 and rra2, which have a reduced residual arabinose content in a polymer tightly associated with the cellulosic wall residue. AB - Two putative glycosyltransferases in Arabidopsis thaliana, designated reduced residual arabinose-1 and -2 (RRA1 and RRA2), are characterized at the molecular level. Both genes are classified in CAZy GT-family-77 and are phylogenetically related to putative glycosyltranferases of Chlamydomonas reinhardtii. The expression pattern of the two genes was analyzed by semi-quantitative RT-PCR using mRNA extracted from various organs of bolting Arabidopsis thaliana plants. In addition, promoter::gusA analysis of transgenic Arabidopsis thaliana containing a fusion between either the RRA-1 or -2 promoter fragment and the gusA reporter gene showed that whereas the RRA1 promoter was primarily active in the apical meristem, the expression pattern of the RRA2 promoter was more diverse but also highly active in the meristematic region. In addition, T-DNA mutant insertion lines of both RRA-1 and -2, were identified and characterized at the molecular and biochemical level. Monosaccharide compositional analyses of cell wall material isolated from the meristematic region showed a ca. 20% reduction in the arabinose content in the insoluble/undigested cell wall residue after enzymatic removal of xyloglucan and pectic polysaccharides. These data indicate that both RRA-1 and -2 play a role in the arabinosylation of cell wall component(s). PMID- 17401636 TI - Body mass index, physical activity and the risk of pancreatic cancer in relation to smoking status and history of diabetes: a large-scale population-based cohort study in Japan--the JPHC study. AB - OBJECTIVE: The effects of BMI and physical activity on the risk of pancreatic cancer were investigated in a large population-based cohort study in Japan (JPHC study). In particular, we explored how these effects were influenced by smoking status and a history of diabetes. METHODS: In total, 99,670 participants (47,499 men, and 52,171 women) were followed for an average of 11 years through the end of 2003. Hazard ratios (HR) and their 95% confidence intervals (CI) were estimated by Cox proportional hazards models. RESULTS: A total of 224 incident cases (128 men, 96 women) of pancreatic cancer were identified. A statistically significant excess risk of pancreatic cancer was associated with current smoking (HR=1.8, CI 1.1-3.0) and a history of diabetes (HR=2.1, CI 1.3-3.5) among men. A similar increase in risk associated with current smoking (HR=2.0, CI 0.9-4.4) and diabetes (HR=1.5, CI 0.7-3.5) was also seen among women. BMI was inversely associated with risk of pancreatic cancer among men, especially among current male smokers or diabetes patients, but no association was found among women. Leisure-time physical activity was not associated with risk in either men or women. CONCLUSIONS: Our study confirms the association between cigarette smoking, history of diabetes and increased risk of pancreatic cancer. However, our data suggest that the association between BMI and risk of pancreatic cancer in this Japanese population may be different from that in Western populations. PMID- 17401637 TI - The future of outcomes measurement: item banking, tailored short-forms, and computerized adaptive assessment. AB - The use of item banks and computerized adaptive testing (CAT) begins with clear definitions of important outcomes, and references those definitions to specific questions gathered into large and well-studied pools, or "banks" of items. Items can be selected from the bank to form customized short scales, or can be administered in a sequence and length determined by a computer programmed for precision and clinical relevance. Although far from perfect, such item banks can form a common definition and understanding of human symptoms and functional problems such as fatigue, pain, depression, mobility, social function, sensory function, and many other health concepts that we can only measure by asking people directly. The support of the National Institutes of Health (NIH), as witnessed by its cooperative agreement with measurement experts through the NIH Roadmap Initiative known as PROMIS (www.nihpromis.org), is a big step in that direction. Our approach to item banking and CAT is practical; as focused on application as it is on science or theory. From a practical perspective, we frequently must decide whether to re-write and retest an item, add more items to fill gaps (often at the ceiling of the measure), re-test a bank after some modifications, or split up a bank into units that are more unidimensional, yet less clinically relevant or complete. These decisions are not easy, and yet they are rarely unforgiving. We encourage people to build practical tools that are capable of producing multiple short form measures and CAT administrations from common banks, and to further our understanding of these banks with various clinical populations and ages, so that with time the scores that emerge from these many activities begin to have not only a common metric and range, but a shared meaning and understanding across users. In this paper, we provide an overview of item banking and CAT, discuss our approach to item banking and its byproducts, describe testing options, discuss an example of CAT for fatigue, and discuss models for long term sustainability of an entity such as PROMIS. Some barriers to success include limitations in the methods themselves, controversies and disagreements across approaches, and end-user reluctance to move away from the familiar. PMID- 17401638 TI - Apoptotic cleavage of NuMA at the C-terminal end is related to nuclear disruption and death amplification. AB - NuMA is a nuclear matrix protein in interphase and distributes to the spindle poles during mitosis. While the essential function of NuMA for mitotic spindle assembly is well established, a structural role of NuMA in interphase nucleus has also been proposed. Several observations suggest that the apoptotic degradation of NuMA may relate to chromatin condensation and micronucleation. Here we demonstrate that four apoptotic cleavage sites are clustered at a junction between the globular tail and the central coiled-coil domains of NuMA. Cleavage of a caspase-6-sensitive site at D(1705) produced the R-form, a major tail-less product of NuMA during apoptosis. The other two cleavage sites were defined at D(1726) and D(1747) that were catalyzed, respectively, by caspase-3 and an unknown aspartase. A NuMA deletion mutant missing the entire cleavage region of residues 1701-1828 resisted degradation and protected cells from nuclear disruption upon apoptotic attack. Under such conditions, cytochrome c was released from mitochondria, but the subsequent apoptotic events such as caspase-3 activation, poly(ADP-ribose) polymerase degradation, and DNA fragmentation were attenuated. Conversely, the tail-less NuMA alone, a mutant mimicking the R-form, induced chromatin condensation and activated the death machinery. It supports that intact NuMA is a structural element in maintaining nuclear integrity. PMID- 17401639 TI - The Doctor by Luke Fildes: an icon in context. AB - This paper discusses one of the most famous paintings on medical themes: The Doctor by Sir Luke Fildes (Fig. 1), which exemplifies how an ideal type of doctoring is construed from reality and from the views and expectations of both the public and doctors themselves. A close reading of The Doctor elucidates three fundamental conflicts in medicine: the first is between statistical efficiency in accordance with scales of morbidity and mortality and the personal devotion that every sick child or suffering individual wants to receive; the second is between the doctor-dominated market and the patient-dominated market; and the third is between influential and rich doctors ("consultants") and practitioners of family medicine (GPs). PMID- 17401640 TI - Genetic mixed linear models for twin survival data. AB - Twin studies are useful for assessing the relative importance of genetic or heritable component from the environmental component. In this paper we develop a methodology to study the heritability of age-at-onset or lifespan traits, with application to analysis of twin survival data. Due to limited period of observation, the data can be left truncated and right censored (LTRC). Under the LTRC setting we propose a genetic mixed linear model, which allows general fixed predictors and random components to capture genetic and environmental effects. Inferences are based upon the hierarchical-likelihood (h-likelihood), which provides a statistically efficient and unified framework for various mixed-effect models. We also propose a simple and fast computation method for dealing with large data sets. The method is illustrated by the survival data from the Swedish Twin Registry. Finally, a simulation study is carried out to evaluate its performance. PMID- 17401641 TI - The state of methods and tools for social systems change. AB - The last decade or so has seen an increase in the rate of attention to systems change and the application of multiple theories to study and influence change in systems. This article considers how these theories, described in other articles in this issues, plus theories about complex adaptive systems open opportunities for us to continue to expand our repertoire of tools and methods to address systems change. These conceptual frameworks, tools, and methods are expected to help us increase our capacity to study and influence both planned and self organizing systems change. New tools and methods have emerged and others are needed to help us continue to explore and influence the complex social systems within which we operate. PMID- 17401642 TI - Effects of positive youth development programs on school, family, and community systems. AB - A review of efforts at social system change in 526 universal competence-promotion outcome studies indicated that 64% of the interventions attempted some type of microsystemic or mesosystemic change involving schools, families, or community based organizations in an attempt to foster developmental competencies in children and adolescents. Only 24% of the reports provided quantitative data on the change that occurred in targeted systems. However, studies containing the necessary information produced several mean effect sizes that were statistically significant, and ranged from modest to large in magnitude. These data indicate that attempts to change social systems affecting children and adolescents can be successful. Future work should measure more thoroughly the extent to which the systemic changes that are targeted through intervention are achieved, and investigate how such changes contribute to the development and sustainability of the outcomes that might be demonstrated by participants of competence-promotion programs. PMID- 17401643 TI - Using system differences to orchestrate change: a systems-guides intervention model. AB - Successful community-based interventions require that change agents give attention to the complexities of interwoven systems, described in the literature as complicated or complex, purposeful or purposive. The author further notes the separate standards, norms, principles and/or goals (here-named systems-guides) of systems. This paper describes the author's experience as a consultant to a multi system collaboration where observed tensions among participants resulted in the author's implementation of a three-step systems-guides model, which can increase change agents' effectiveness with systems. The steps comprise: detection (of systems, systems-guides, and discrepancies between them; here through observation); documentation (of discrepancies; here through small group discussions); dissemination (to systems participants; here to the collaboration). Two small but fundamental systems changes emerged from systems participants, not from the change agent/author who facilitated systems members' recognition of discrepancies. A major limitation in the implementation of this systems-guides model centers on the critical necessity of the close relationship change agents must have with the involved systems. PMID- 17401644 TI - Attachment style, home-leaving age and behavioral problems among residential care children. AB - In a prospective study, the attachment style, home-leaving age, length of time in residential care, and behavioral problems among Israeli residential care children (N = 68), were studied. Data analyses showed that children removed from their homes at a later age suffered from higher levels of anxiety, depression and social problems compared to children taken from their homes at the age of 7 or less. It was also found that a prolonged stay (over 2 years) in residential care was related to higher levels of anxiety and depression. An interaction effect of a child's attachment style and home-leaving age was found in the child's level of aggressive behavior. PMID- 17401645 TI - Differential impacts of public health insurance expansions at the local level. AB - Dramatic expansions in public health insurance eligibility for U.S. children have only modestly reduced the aggregate number of uninsured at the national level. This paper shows that Medicaid and SCHIP expansions had different impacts on child health insurance coverage patterns based upon local labor market characteristics. Metropolitan areas with high levels of unemployment were most likely to have seen improvements in overall insurance coverage for children between 1990 and 2001. Areas with greater fractions of employment in services, retail or wholesale trade were more likely to have experienced increases in public coverage but not overall coverage rates. PMID- 17401646 TI - Temporal profiles of cytoskeletal protein loss following traumatic axonal injury in mice. AB - To examine the time course and relative extent of proteolysis of neurofilament and tubulin proteins after traumatic axonal injury (TAI), anesthetized mice were subjected to optic nerve stretch injury. Immunohistochemistry confirmed neurofilament accumulation within axonal swellings at 4, 24, and 72 h postinjury (n = 4 injured and 2 sham per time point). Immunoblotting of optic nerve homogenates (n = 5 injured and 1 sham at 0.5, 4, 24 or 72 h) revealed calpain mediated spectrin proteolytic fragments after injury. Protein levels for NF68 progressively decreased from 0.5 h to 24 h postinjury, while NF200 and alpha tubulin levels decreased acutely (0.5-4 h), with a secondary decline at 72 h postinjury. These data demonstrate that diffusely distributed TAI is associated not only with a localized accumulation of neurofilament proteins, but also significant decreases in total cytoskeletal protein levels which may be mediated, in part, by calpains. Protection of the axonal cytoskeleton represents a potential therapeutic target for axonal damage associated with injury or neurodegenerative diseases. PMID- 17401647 TI - Distribution of immunoreactive prolyl oligopeptidase in human and rat brain. AB - Prolyl oligopeptidase (POP) is a serine endoprotease that hydrolyses peptides shorter than 30-mer. POP may have a role in inositol 1,4,5-triphosphate (IP(3)) signaling and in the actions of antidepressants, and POP inhibitors have exhibited antiamnesic and neuroprotective properties. However, little is known about the distribution of POP protein in the brain. We used immunohistochemistry to localize POP enzyme in the human whole hemisphere and in the rat whole brain. In humans, the highest POP densities were observed in caudate nucleus and putamen, hippocampus and cortex. In the rat, the highest POP densities were found in substantia nigra, hippocampus, cerebellum and caudate putamen. In general, the distribution of POP in human and rat brains was very similar and resembled that of IP(3) receptors. Our findings are support for a role of POP in movement regulation, cognition and possibly in IP(3) signaling. The expression of POP in processing nuclei further supports its function beyond neuropeptide metabolism. PMID- 17401648 TI - Modulation of [3H]dopamine release by glutathione in mouse striatal slices. AB - Glutathione (gamma-glutamylcysteinylglycine, GSH and oxidized glutathione, GSSG), may function as a neuromodulator at the glutamate receptors and as a neurotransmitter at its own receptors. We studied now the effects of GSH, GSSG, glutathione derivatives and thiol redox agents on the spontaneous, K(+)- and glutamate-agonist-evoked releases of [(3)H]dopamine from mouse striatal slices. The release evoked by 25 mM K(+) was inhibited by GSH, S-ethyl-, -propyl-, -butyl and pentylglutathione and glutathione sulfonate. 5,5'-Dithio-bis-2-nitrobenzoate (DTNB) and L-cystine were also inhibitory, while dithiothreitol (DTT) and L cysteine enhanced the K(+)-evoked release. Ten min preperfusion with 50 microM ZnCl(2) enhanced the basal unstimulated release but prevented the activation of K(+)-evoked release by DTT. Kainate and 2-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) evoked dopamine release but the other glutamate receptor agonists N-methyl-D-aspartate (NMDA), glycine (1 mM) and trans-1 aminocyclopentane-1,3-dicarboxylate (t-ACPD, 0.5 mM), and the modulators GSH, GSSG, glutathione sulfonate, S-alkyl-derivatives of glutathione, DTNB, cystine, cysteine and DTT (all 1 mM) were without effect. The release evoked by 1 mM glutamate was enhanced by 1 mM GSH, while GSSG, glutathionesulfonate and S-alkyl derivatives of glutathione were generally without effect or inhibitory. NMDA (1 mM) evoked release only in the presence of 1 mM GSH but not with GSSG, other peptides or thiol modulators. L-Cysteine (1 mM) enhanced the glutamate-evoked release similarly to GSH. The activation by 1 mM kainate was inhibited by S-ethyl , -propyl-, and -butylglutathione and the activation by 0.5 mM AMPA was inhibited by S-ethylglutathione but enhanced by GSSG. Glutathione alone does not directly evoke dopamine release but may inhibit the depolarization-evoked release by preventing the toxic effects of high glutamate, and by modulating the cysteine cystine redox state in Ca(2+ )channels. GSH also seems to enhance the glutamate agonist-evoked release via both non-NMDA and NMDA receptors. In this action, the gamma-glutamyl and cysteinyl moieties of glutathione are involved. PMID- 17401649 TI - Age-related oxidative modifications of proteins and lipids in rat brain. AB - Oxidants have been shown to play a major role in ageing and ageing-related neurodegenerative diseases. In the present study, we investigated the effect of ageing on oxidative damage to lipids and proteins in brain homogenate, mitochondria and synaptosomes of adult (6-month-old), old (15-month-old), and senescent (26-month-old) Wistar rats. There was a significant increase in thiobarbituric acid-reactive substances and conjugated dienes in homogenates, which indicate increased lipid peroxidation (LPO). Oxidative modifications of homogenate proteins were demonstrated by a loss of sulfhydryl content, accumulation of dityrosines and formation of protein conjugates with LPO-end products. Increase in protein conjugates with LPO-end products and a decrease in SH groups were observed also in mitochondria and synaptosomes, but dityrosine content was elevated only in synaptosomes. Protein surface hydrophobicity, measured by fluorescent probe 1-anilino-8-naphthalenesulfonate (ANS), was increased only in homogenate. These results suggest that besides mitochondria and synaptosomes other cellular compartments are oxidatively modified during brain ageing. PMID- 17401650 TI - Effects of chronic haloperidol and/or clozapine on oxidative stress parameters in rat brain. AB - Decreased antioxidant activity is considered as one of the causes of tardive dyskinesia in schizophrenic patients in a prolonged neuroleptic treatment course. Haloperidol (HAL) has been hypothesized to increase oxidative stress, while clozapine (CLO) would produce less oxidative damage. The objective was to determine whether CLO for 28 days could reverse or attenuate HAL-induced oxidative damage in animals previously treated with HAL for 28 days. HAL significantly increased thiobarbituric acid reactive substances levels in the cortex (CX) and striatum and increased protein carbonyls in hippocampus (HP) and CX and this was not attenuated by CLO treatment. In the total radical trapping antioxidant parameter assay there was a decrease in the HP total antioxidant potential induced by HAL and by treatment with HAL + CLO. Our findings demonstrated that the atypical antipsychotic CLO could not revert oxidative damage caused by HAL. PMID- 17401651 TI - Caspase-dependent apoptosis induced by thapsigargin was prevented by glycogen synthase kinase-3 inhibitors in cultured rat cortical neurons. AB - Calcium ion is essential for cellular functions including signal transduction. Uncontrolled calcium stress has been linked causally to a variety of neurodegenerative diseases. Thapsigargin, which inhibits Ca(2+)-ATPase in the endoplasmic reticulum (ER) and blocks the sequestration of calcium by the ER, induced apoptotic cell death (chromatin condensation and nuclear fragmentation) accompanied by GRP78 protein expression and caspase-3 activation in rat fetal cortical neurons (days in vitro 9-10). Blockade of N-methyl-D-aspartate (NMDA) receptors with NMDA antagonists induced apoptosis without GRP78 protein expression. Apoptosis accompanied both caspase-9 and caspase-3 activation. We then examined whether GSK-3 is involved in thapsigargin-induced cell death by using GSK-3 inhibitors. We assayed the effects of selective GSK-3 inhibitors, SB216763, alsterpaullone and 1-azakenpaullone, on thapsigargin-induced apoptosis. These inhibitors completely protected cells from thapsigargin-induced apoptosis. In addition, GSK-3 inhibitors inhibited caspase-9 and caspase-3 activation accompanied by thapsigargin-induced apoptosis. These results suggest that thapsigargin induces caspase-dependent apoptosis mediated through GSK-3beta activation in rat cortical neurons. PMID- 17401652 TI - Inhibition of melatonin biosynthesis induces neurofilament hyperphosphorylation with activation of cyclin-dependent kinase 5. AB - Decreased level of melatonin and hyperphosphorylation of neurofilament proteins have been reported in Alzheimer's disease (AD). However, the direct evidence linking melatonin and neurofilament phosphorylation is still lacking. Here, we investigated the effect of inhibiting melatonin biosynthesis on phosphorylation of neurofilament proteins and the involvement of cyclin-dependent kinase 5 (cdk 5) in rats. We observed that injection of haloperidol, a specific inhibitor of 5 hydroxyindole-O-methyltransferase, resulted in significantly decreased level of serum melatonin with a concomitantly increased phosphorylation of neurofilament proteins and activation of cdk-5 in rats. Exogenous supplementation of melatonin partially arrested the hyperphosphorylation of neurofilament and the activation of cdk-5. These results suggest that inhibition of melatonin biosynthesis may activate cdk-5 and thus induces Alzheimer-like hyperphosphorylation of neurofilament proteins. PMID- 17401653 TI - Effect of resuscitation with 21% oxygen and 100% oxygen on NMDA receptor binding characteristics following asphyxia in newborn piglets. AB - The present study investigated the effect of reventilation with 21% and 100% oxygen following asphyxia in newborn piglets on NMDA receptor binding characteristics, Na(+), K(+)-ATPase activity, and lipid peroxidation. After achieving a heart rate less than 60 beats per minute, asphyxiated piglets were reventilated with 21% oxygen or 100% oxygen. (3)[H]MK-801 binding showed the Bmax in the 21% and 100% groups to be 1.53 +/- 0.43 and 1.42 +/- 0.35 pmol/mg protein (p = ns). Values for Kd were 4.56 +/- 1.29 and 4.17 +/- 1.05 nM (p = ns). Na(+), K(+)-ATPase activity in the 21% and 100% groups were 23.5 +/- 0.9 and 24.4 +/- 3.9 micromol Pi/mg protein/h (p = ns). Conjugated dienes (0.05 +/- 0.02 vs. 0.07 +/- 0.03 micromol/g brain) and fluorescent compounds (0.54 +/- 0.05 vs. 0.78 +/- 0.19 microg quinine sulfate/g brain), were similar in both groups (p = ns). Though lipid peroxidation products trended higher in the 100% group, these data show that NMDA receptor binding and Na(+), K(+)-ATPase activity were similar following reventilation with 21% or 100% oxygen after a single episode of mild asphyxia. PMID- 17401654 TI - AT1 receptor blocker candesartan-induced attenuation of brain injury of rats subjected to chronic cerebral hypoperfusion. AB - One of common pathophysiological states associated with central nervous system is chronic cerebral hypoperfusion (CH) that frequently occurs in conditions such as vascular dementia and Alzheimer's disease. Long term blockage of angiotensin II type 1 (AT(1)) receptor provides protection from ischemia induced injury of brain as well as reduction of cerebrovascular inflammation. Examining effect of the blockage on reduced glutathione (GSH), ascorbic acid (AA), and lipid peroxidation were of purpose in the present study. Modeling CH, rats were subjected to permanent occlusion of common carotid arteries bilaterally. AT(1 )receptor antagonist, candesartan, was given daily for 14 days after surgery. CH caused a significant increase in lipid peroxidation and decrease in GSH content of cerebral hippocampal tissue with no change in AA level. Candesartan (0.5 mg/kg, oral) not only reduced lipid peroxidation but also restored GSH significantly besides elevating AA and improving histopathological alterations. In conclusion, long term AT(1 )receptor blockage may be considered as novel therapeutic approach for protection from damage associated with CH. Underlying mechanism(s) may in part be related to suppressing oxidative stress and preserving brain antioxidant capacity. PMID- 17401655 TI - Neuroprotective effect of ganglioside GM1 on the cytotoxic action of hydrogen peroxide and amyloid beta-peptide in PC12 cells. AB - Ganglioside GM1 was shown to increase the viability of PC12 cells exposed to hydrogen peroxide or amyloid beta-peptide (Abeta(25-35)). The PC12 cells transfected with mutant gene (expressing APP(SW)) were found to be more sensitive to oxidative stress than the cells transfected with wild type gene (expressing APP(WT)) or vector-transfected cells, GM1 being effective in enhancing the viability of the cells transfected with mutant gene. The exposure to hydrogen peroxide or Abeta(25-35) results in a partial inactivation of Na(+),K(+)-ATPase in PC12 cells, H(2)O(2) increases MDA accumulation in these cells. But these effects could be partially prevented or practically abolished by GM1 ganglioside. In the presence of the inhibitor of tyrosine kinase of Trk receptors (K-252a) the protective and metabolic effects of GM1 on PC12 cells in conditions of oxidative stress caused by hydrogen peroxide are not observed or are markedly diminished. PMID- 17401656 TI - Effects of phenylalanine and its metabolites on cytoplasmic free calcium in cortical neurons. AB - Classic phenylketonuria (PKU) is characterized by brain lesions. However, its underlying neurotoxic mechanisms remain unknown. Based on our previous studies, we hypothesized that calcium might participate in PKU-associated neuropathy. In cultured cortical neurons, cytoplasmic free calcium concentration ([Ca(2+)](i)) decreased dramatically when treatment with phenylalanine (Phe) and phenyllactic acid, while phenylacetic acid treatment immediately increased [Ca(2+)](i), which began to decrease after 3 min. Moreover, [Ca(2+)](i) decreased dramatically after Phe treatment in the presence of EGTA suggesting that Phe might increase [Ca(2+)](i) efflux. Phe-induced [Ca(2+)](i) decrease was strongly inhibited by vanadate, a non-specific plasma membrane Ca(2+)-ATPase (PMCA) antagonist, suggesting that Phe might increase [Ca(2+)](i) efflux throught modulating PMCA. These findings were further supported by the facts that Phe could increase membrance (45)Ca-uptake capability and PMCA activity. In contrast, treatment of KBR7943 or thapsigargin, antagonists to Na/Ca Exchanger (NCX) and Sarco/Endoplasmic reticulum Ca(2+)-ATPase (SERCA), respectively, did not elicit any changes in [Ca(2+)](i). Specific siRNA against PMCA had an effect similar to vanadate. Since the brain injury induced by phenylalaninemia was thought to be a chronic process, we cultured cortical neurons in the presence of Phe for 2 weeks and measured [Ca(2+)](i), PMCA activity and (45)Ca-uptake capability at days 3, 7, 9 and 14, respectively. PMCA activity and (45)Ca-uptake capability decreased from day 9, at the same time [Ca(2+)](i) increase was observed. In conclusion, PMCA participate in regulating Phe-induced initial rapid decrease in [Ca(2+)](i) and subsequent long-term increase in [Ca(2+)](i). PMID- 17401657 TI - The study of Golgi apparatus in Alzheimer's disease. AB - Alzheimer's disease is an irreversible, progressive neurodegenerative disorder leading invariably to death, usually within 7-10 years after diagnosis and is the leading cause of dementia in the elderly. Not only is Alzheimer's disease a tragic disease in which people suffer from neurodegeneration in the years to come, it also becomes an incredible burden on the public health system. However, there is currently no effective treatment to halt the progression or prevent the onset of Alzheimer's disease. This is partly due to the fact that the complex pathophysiology of Alzheimer's disease is not yet completely understood. Recently, Golgi apparatus is found to play an important role in Alzheimer's disease. In this review, we discuss the changes of Golgi apparatus during clinical progression and pathological development of Alzheimer's disease. First, changes of Golgi apparatus size in Alzheimer's disease are summarized. We then address the role of Golgi apparatus in the neuropathology of Alzheimer's disease. Finally, the role of Golgi apparatus in the pathogenesis of Alzheimer's disease is discussed. Understanding the contribution of Golgi apparatus dysfunction to Alzheimer's disease and its pathophysiological basis will significantly impact our ability to develop more effective therapies for Alzheimer's disease. PMID- 17401658 TI - Differential expression of apoptotic proteins following hypoxia-induced CREB phosphorylation in the cerebral cortex of newborn piglets. AB - The present study investigates the correlation between the hypoxia-induced phosphorylation of cyclic AMP response element binding protein and the expression of apoptotic proteins (proapoptotic proteins Bax and Bad and antiapoptotic proteins Bcl-2 and Bcl-xl) during hypoxia in the cerebral cortex of newborn piglets. Piglets were divided into normoxic (Nx) and hypoxic (Hx, FiO(2)=0.06 for 1 h) groups. Cerebral tissue hypoxia was documented by ATP and phosphocreatine (PCr) levels. Ser(133) phosphorylation of cyclic AMP response element binding (CREB) protein was determined by Western blot analysis using a specific anti phosphorylated Ser(133)-CREB protein antibody. The expression of apoptotic proteins was determined by using specific anti-Bax, anti-Bad, anti-Bcl-2 and anti Bcl-xl antibodies. ATP and PCr values (mumoles/g brain) in Hx were significantly different from Nx (ATP: 4.40 +/- 0.39 in Nx vs. 1.19 +/- 0.44 in Hx, P<0.05 vs. Nx; PCr: 3.60 +/- 0.40 in Nx vs. 0.70 +/- 0.31 in Hx, P<0.05 vs. Nx). Ser(133) phosphorylated CREB protein (OD x mm(2)) was 74.55 +/- 4.75 in Nx and 127.13 +/- 19.36 in Hx (P<0.05 vs. Nx). The expression of proapoptotic proteins Bax and Bad increased and strongly correlated with the increase in CREB protein phosphorylation (correlation coefficient r=0.82 and r=0.85, respectively). The expression of antiapoptotic proteins Bcl-2 and Bcl-xl did not show correlation with CREB protein phosphorylation. We conclude that cerebral hypoxia results in differential regulation of CREB protein-mediated expression of proapoptotic and antiapoptotic proteins in the cerebral cortex of newborn piglets. We propose that the increased expression of proapoptotic vs antiapoptotic genes will lead to an increased potential for apoptotic programmed cell death in the Hx newborn brain. PMID- 17401659 TI - In vitro studies on the putative function of N-acetylaspartate as an osmoregulator. AB - Efflux and tissue content of N-acetylaspartate (NAA) and amino acids were evaluated from cultured and acutely prepared hippocampal slices in response to changes in osmolarity. The osmoregulator taurine, but not NAA, was lost from both types of slices after moderate reductions in extracellular osmolarity (-60 mOsm) for 10-48 h. Hypoosmotic shock (-166 mOsm) for 5 min resulted in unselective efflux of several amino acids from acutely prepared slices. Notably, the efflux of taurine, but not NAA, was prominent also after the shock. Efflux of NAA was markedly enhanced by NMDA and high K(+), in particular after the stimulation period. The high K(+)-mediated efflux was decreased by high extracellular osmolarity and a NMDA-receptor antagonist. The results indicate that NAA efflux can be induced by a sudden non-physiological decrease in extracellular osmolarity but not by prolonged more moderate changes in osmolarity. The mechanisms behind the efflux of NAA by high K(+) are complex and may involve both swelling and activation of NMDA-receptors. PMID- 17401660 TI - Apoptotic mode of cell death in substantia nigra following intranigral infusion of the parkinsonian neurotoxin, MPP+ in Sprague-Dawley rats: cellular, molecular and ultrastructural evidences. AB - The potent parkinsonian neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6 tetrahydropyridine (MPTP) is known to cause dopaminergic neurodegeneration in nigrostriatal system. In the present study we investigated the nuclear morphology of cells in the substantia nigra pars compacta (SNpc) region of rats following unilateral intranigral infusion of the active metabolite, 1-methyl-4-phenyl pyridinium ion (MPP(+)), which resulted in a dose-dependent and prolonged dopamine depletion in the ipsilateral striatum. There appeared a substantial loss of tyrosine hydroxylase immunoreactive neurons in the SNpc that received the neurotoxin. Specific nuclear staining with Hoechst 33342 or acridine orange revealed bright pyknotic, shrunken, distorted nuclei and condensed chromatin with perinuclear nucleolus respectively following visualization with the former and latter dyes in the ipsilateral SNpc, as compared to the round, intact nuclei and centrally positioned nucleolus in the contralateral side. Ultrastructural details of the nucleus under transmission electron microscope confirmed distorted nuclear organization with shrunken or condensed nuclei and disrupted nuclear membrane. These features are typical of nucleus undergoing apoptosis, and suggest that MPP(+) causes dopaminergic neuronal death through an apoptotic mode. Typical laddering pattern of genomic DNA isolated from the ipsilateral SN in agarose gel electrophoresis conclusively established apoptosis following intranigral administration of MPP(+) in rats. PMID- 17401661 TI - NTPDase and 5'-nucleotidase activities of synaptosomes from hippocampus of rats subjected to hyperargininemia. AB - ATP is an important excitatory neurotransmitter and adenosine acts as a neuromodulatory structure inhibiting neurotransmitters release in the central nervous system. Since the ecto-nucleotidase cascade that hydrolyzes ATP to adenosine is involved in the control of brain functions and previous studies realized in our laboratory have recently reported that acute administration of Arg decreases the NTPDase and 5'-nucleotidase activities of rat blood serum, in the present study we investigated the effect of arginine administration on NTPDase and 5'-nucleotidase activities by synaptosomes from hippocampus of rats. First, sixty-days-old rats were treated with a single or a triple intraperitoneal injection of arginine (0.8 g/Kg) or an equivalent volume of 0.9% saline solution (control) and were killed 1 h later. Second, rats received an intracerebroventricular injection of 1.5 mM arginine solution or saline (5 microL) and were killed 1 h later. We also tested the in vitro effect of arginine (0.1-1.5 mM) on nucleotide hydrolysis in synaptosomes from rat hippocampus. Results showed that intraperitoneal arginine administration did not alter nucleotide hydrolysis. On the other hand, arginine administered intracerebroventricularly reduced ATP (32%), ADP (30%) and AMP (21%) hydrolysis, respectively. In addition, arginine added to the incubation medium, provoked a decrease on ATP (19%), ADP (17%) and AMP (23%) hydrolysis, respectively. Furthermore, kinetic studies showed that the inhibitory effect of arginine was uncompetitive in relation to ATP, ADP and AMP. In conclusion, according to our results it seems reasonable to postulate that arginine alters the cascade involved in the extracellular degradation of ATP to adenosine. PMID- 17401662 TI - Protective effects of lamotrigine, aripiprazole and escitalopram on depression induced oxidative stress in rat brain. AB - We investigated the effects of lamotrigine, aripiprazole and escitalopram administration and experimental depression on lipid peroxidation (LP) and antioxidant levels in cortex of the brain in rats. Forty male wistar rats were randomly divided into five groups. First group was used as control although second group was depression-induced group. Aripiprazole, lamotrigine and escitalopram per day were orally supplemented to chronic mild stress (CMS) depression-induced rats constituting the third, fourth and fifth groups for 28 days, respectively. Depression resulted in significant decrease in the glutathione peroxidase (GSH-Px) activity, reduced glutathione and vitamin C of cortex of the brain although their levels and beta-carotene concentrations were increased by the three drugs administrations to the animals of CMS induced depression group. The LP levels in the cortex of the brain and plasma of depression group were elevated although their levels were decreased by the administrations. The increases of antioxidant values in lamotrigine group were higher according to aripiprazole and escitalopram supplemented groups. Vitamin A level did not change in the five groups. In conclusion, the experimental depression is associated with elevated oxidative stress although treatment with lamotrigine has most protective effects on the oxidative stress within three medicines. PMID- 17401663 TI - Erythropoietin in cerebrospinal fluid: age-related reference values and relevance in neurological disease. AB - We aimed to establish age-related reference values for Erythropoietin (EPO) in cerebrospinal fluid (CSF) and to evaluate concentrations in neurological diseases. CSF and serum EPO was measured in controls with tension-type headache (CTTH), in patients with ALS, dementia and depression using ELISA technique. Stability experiments showed CSF EPO to be stable for two and a half months and over two thaw/freeze cycles. A positive correlation of CSF EPO with age was found (P<0.01). We found a CSF/serum EPO concentration ratio of 0.126, pointing towards an intrathecal synthesis of EPO. The ALS group showed significantly lowered CSF EPO compared to age-matched CTTH (P<0.012), whereas the dementia and depression group showed no significant differences compared to CTTH.The establishment of age related reference values in a large cohort of controls will improve the interpretation of future CSF EPO evaluations in neurological diseases. PMID- 17401664 TI - Oxidative stress in rat retina and hippocampus after chronic MDMA ('ecstasy') administration. AB - The effects of MDMA administration on oxidative stress markers in rat eye and hippocampus, and the neuroprotective effects of the antioxidant 3,4-dihydro-6 hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran (CR-6) have been studied. MDMA effects on liver were used for comparison with those in eye and hippocampus and to test CR-6 protective effects. Another goal was to test for apoptosis in retinal cells, as it is known that happens in liver and brain. After 1 week of ecstasy administration, malondialdehyde (MDA) concentration increased, glutathione peroxidase (GPx) activity and glutathione (GSH) content decreased in liver, as previously described. MDA concentration increased and GPx activity decreased in hippocampus; whereas no change was observed in GSH concentration. MDMA decreased ocular GSH concentration and GPx activity; no change was observed in MDA concentration. The number of TUNEL-positive nuclei increased significantly in rat retinas after 1 week of MDMA administration. CR-6 normalized the modifications in liver, hippocampus and retina mentioned above. PMID- 17401665 TI - Effect of radicicol infusion on the Src tyrosine kinase activity of rat hippocampus before and after training in an inhibitory avoidance task. AB - The participation of protein serine/threonine kinases in memory formation and retrieval is well established. In contrast, relatively little is known on the role of protein tyrosine kinases (PTKs). Previous work showed that intra hippocampal infusion of the Src-PTK inhibitor radicicol inhibits memory acquisition, consolidation, and retrieval of one-trial step-down inhibitory avoidance task. In this study, we investigated the possible interaction between levels of Src-PTK activity in hippocampus and memory acquisition, formation, and retrieval of this task. Radicicol (0.5 microg/ml) was infused into the CA1 region of the hippocampus of rats trained in a one-trial step-down inhibitory avoidance task. Radicicol infused 15 min before training decreased Src-PTK activity, as measured 0, 1.5, and 24 h after training, and impaired memory acquisition of the task. When given immediately after training, there was a decrease in Src-PTK activity 1.5 h, but not 0 or 24 h after training. This treatment depressed memory consolidation. Radicicol infused into CA1 10 min prior to retrieval testing inhibited hippocampal Src-PTK activity, as measured immediately after the test session. The results suggest that Src-PTKs participate in memory acquisition, consolidation, and retrieval processes, but the timing of the role of the enzyme is different in each case. PMID- 17401666 TI - Effect of hyperoxia on cortical neuronal nuclear function and programmed cell death mechanisms. AB - There is growing concern over detrimental neurologic effects to human newborns caused by increased inspired oxygen concentrations. We hypothesize that hyperoxia (FiO(2)>0.95) results in increased high-affinity Ca(2+)-ATPase activity, Ca(2+) influx, and proapoptotic protein expression in cortical neuronal nuclei of newborn piglets. Neuronal cerebral energy metabolism was documented by determining ATP and phosphocreatine levels. Neuronal nuclear conjugated dienes and fluorescent compounds were measured as indices of lipid peroxidation. High affinity Ca(2+)-ATPase activity and ATP-dependent Ca(2+)-influx were determined to document neuronal nuclear membrane function. Hyperoxia resulted in increases in lipid peroxidation, high-affinity Ca(2+)-ATPase activity, ATP-dependent Ca(2+) influx, and Bax/Bcl-2 ratio in the cortical neuronal nuclei of newborn piglets. We conclude that hyperoxia results in modification of neuronal nuclear membrane function leading to increased nuclear Ca(2+)-influx, and propose that hyperoxia induced increases in intranuclear Ca(2+) activates the Ca(2+)/calmodulin dependent protein kinase pathway, triggering increased CREB protein-mediated apoptotic protein expression in hyperoxic neurons. PMID- 17401667 TI - c-Myb immunoreactivity, protein and mRNA levels significantly increase in the aged hippocampus proper in gerbils. AB - Myb genes are a family of transcription factors and have been implicated in the control of the proliferation and differentiation of normal and transformed cells. c-Myb is the best characterized member of the myb family. In the present study, we investigated age-dependent changes of c-myb immunoreactivity, its protein and mRNA level in the hippocampus proper (CA1-3 regions) at various age stages in gerbils. In the postnatal month 1 (PM 1) group, c-myb immunoreactivity was detected in non-pyramidal neurons of the strata oriens and radiatum as well as in pyramidal neurons of the stratum pyramidale. At PM 3, c-myb immunoreactivity and its protein level were similar to those at PM 1. Thereafter, c-myb immunoreactivity and its protein level were increased with time. In the PM 24 group, c-myb immunoreactivity, its protein and mRNA levels were highest. These results suggest that the significant increase of c-myb immunoreactivity, protein and mRNA levels in the aged hippocampus may be associated with neuronal aging. PMID- 17401668 TI - Expression and developmental regulation of the cystine/glutamate exchanger (xc-) in the rat. AB - The cystine/glutamate exchanger (antiporter x (c) (-) ) is a membrane transporter involved in the uptake of cystine, the rate-limiting amino acid in the synthesis of glutathione. Recent studies suggest that the antiporter plays a role in the slow oxidative excitotoxity and in the pathological effects of beta-N-oxalylamino L: -alanine, the molecule responsible for neurolathyrism, a neurotoxic upper motor neuron disease. The mouse cystine/glutamate exchanger has been cloned and showed to be composed of two distinct proteins, one of which being a novel protein, named xCT, of 502 amino acids and 12 putative trans-membrane domains. We have generated and purified a polyclonal antibody to mouse xCT and studied its expression in rat brain and in different cultured cells (astrocytes, fibroblasts and neurons) using Western blot and immunocytochemical techniques. Expression of xCT was also studied in rat brain and muscle at different developmental stages. Parallel experiments were carried out with antibodies to the heavy chain of 4F2 surface antigen, the non-specific subunit of the antiporter x (c) (-) . xCT antibody detected in all cell and tissue extracts a specific band of about 40 kDa. Subcellular fractionation demonstrated that xCT is concentrated mainly in the microsomal-mitochondrial fraction, in accord with its structure as transmembrane protein. Immunocytochemical analysis showed a strong staining in all cells examined, included neurons. Furthermore, both xCT and the heavy chain of 4F2 surface antigen increased in the brain during development, reaching the highest expression in adulthood. The study of the expression and developmental profile of xCT represents a first step towards a better characterization of its biochemical properties and function, which in turn may help to understand the relative contribution of the x (c) (-) antiporter in the pathogenesis of certain neurodegenerative diseases. PMID- 17401669 TI - Anaerobic glycolysis protection against 1-methy-4-phenylpyridinium (MPP+) toxicity in C6 glioma cells. AB - The neurotoxin 1-methy-4-phenylpyridinium (MPP(+)) is used for its' capacity to induce Parkinsonism through its inhibitory effects on mitochondrial complex I. This inhibition disrupts cellular energy formation and aerobic glycolysis. The objective of this study was to demonstrate that the toxic effect of mitochondrial aerobic pathway inhibition with MPP(+ )can be reduced by stimulating anaerobic glycolysis using glucose supplementation. In this study, C6 Glioma cell viability was examined in the presence of different concentrations of MPP alone and with the addition of glucose. The results obtained indicate that there was a significant increase (P < 0.001) in cell viability in cells treated with glucose and MPP(+ )verses cells treated with MPP(+ )alone. Fluorometric analysis using 100 microM Rhodamine 123 indicated mitochondrial membrane potential was not restored in MPP(+ )treated cells with glucose; however, normal cell viability was confirmed using 2 microg/ml Fluorescein diacetate. This dual fluorescence indicated mitochondrial damage from MPP(+ )while glucose augmented cell survival. Further confirmation of cell survival upon damage to the mitochondria was evident in TUNEL staining. Positive staining was prominent only in MPP(+) treatment groups alone, while control and co-treated groups exhibited little to no TUNEL staining. ATP measurements of all MPP(+) treated groups exhibited a significant (P < 0.001) decrease verses control. Groups co-treated with MPP(+ )and glucose revealed a significant increase (250 microM group: P < 0.001) in ATP. It was concluded from this study that glucose supplementation was able to sustain cellular viability and ATP production through anaerobic glycolysis despite the inhibitory effect of MPP(+ )on aerobic glycolysis. PMID- 17401670 TI - Differential expression of cerebellar metabotropic glutamate receptors mGLUR2/3 and mGLUR4a after the administration of a convulsant drug and the adenosine analogue cyclopentyladenosine. AB - Metabotropic glutamate receptors (mGluR) play a role in synaptic transmission, neuronal modulation and plasticity but their action in epileptic activity is still controversial. On the other hand adenosine acts as a neuromodulator with endogenous anticonvulsive properties. Since cerebellum from epileptic patients has shown neuronal damage, sometimes associated with Purkinje cells loss, we have explored the effect of repetitive seizures on two types of mGluR in the cerebellum. Seizures were induced by the convulsant drug 3-mercaptopropionic acid (MP) and the effect of the adenosine analogue cyclopentyladenosine (CPA) alone or before MP administration (CPA+MP) were also evaluated. The expression of the receptors subtypes 2/3 (mGluR2/3) and 4a (mGluR4a) was assessed by immunocitochemistry. Granular cell layer was labeled with mGluR2/3 antibody and increased immunoreactivity was observed after MP (60%), CPA (53%) and CPA + MP (85%) treatments. Control cerebellum slices showed mGluR4a reactivity around Purkinje cells, while MP, CPA and CPA+MP treatment decreased this immunostaining. Repetitive administration of MP and CPA induces an increased cerebellar mGluR2/3 and a decreased mGluR4a immunostaining, suggesting a distinct participation of both receptors that may be related to the type of cell involved. A protective action and /or an apoptotic effect may not be discarded. CPA repetitive administration although increase seizure latency, cannot prevent seizure activity. PMID- 17401671 TI - Endogenous expression of adenosine A1, A2 and A3 receptors in rat C6 glioma cells. AB - Inhibitory and stimulatory adenosine receptors have been identified and characterized in both membranes and intact rat C6 glioma cells. In membranes, saturation experiment performed with [(3)H]DPCPX, selective A(1)R antagonist, revealed a single binding site with a K (D) = 9.4 +/- 1.4 nM and B (max) = 62.7 +/- 8.6 fmol/mg protein. Binding of [(3)H]DPCPX in intact cell revealed a K (D) = 17.7 +/- 1.3 nM and B (max )= 567.1 +/- 26.5 fmol/mg protein. On the other hand, [(3)H]ZM241385 binding experiments revealed a single binding site population of receptors with K (D) = 16.5 +/- 1.3 nM and B (max) = 358.9 +/- 52.4 fmol/mg protein in intact cells, and K (D) = 4.7 +/- 0.6 nM and B (max) = 74.3 +/- 7.9 fmol/mg protein in plasma membranes, suggesting the presence of A(2A) receptor in C6 cells. A(1), A(2A), A(2B) and A(3 )adenosine receptors were detected by Western-blotting and immunocytochemistry, and their mRNAs quantified by real time PCR assays. Gialpha and Gsalpha proteins were also detected by Western-blotting and RT-PCR assays. Furthermore, selective A(1)R agonists inhibited forskolin- and GTP-stimulated adenylyl cyclase activity and CGS 21680 and NECA stimulated this enzymatic activity in C6 cells. These results suggest that C6 glioma cells endogenously express A(1) and A(2) receptors functionally coupled to adenylyl cyclase inhibition and stimulation, respectively, and suggest these cells as a model to study the role of adenosine receptors in tumoral cells. PMID- 17401672 TI - Antiepileptic drugs prevent changes induced by pilocarpine model of epilepsy in brain ecto-nucleotidases. AB - Ecto-nucleotidases, one of the main mechanisms involved in the control of adenosine levels in the synaptic cleft, have shown increased activities after the pilocarpine model of epilepsy. Here we have investigated the effect of the antiepileptic drugs (AEDs) on ecto-nucleotidase activities from hippocampal and cerebral cortical synaptosomes of rats at seven days after the induction of the pilocarpine model. Expression of these enzymes were investigated as well. Our results have demonstrated that phenytoin (50 mg/kg) and carbamazepine (30 mg/kg) were able to prevent the increase in ecto-nucleotidase activities elicited by pilocarpine in brain synaptosomes. However, sodium valproate (at 100 mg/kg) was only able to avoid the increase on ATP and ADP hydrolysis in hippocampal synaptosomes. Increase on ATP hydrolysis in hippocampal synaptosomes was also prevented by sodium valproate at 286 mg/kg, which corresponds to ED50 for pilocarpine model. NTPDase1, NTPDase2, NTPDase3, and ecto-5'-nucleotidase expressions were not affected by pilocarpine in cerebral cortex. However, expressions of NTPDase2, NTPDase3, and ecto-5'-nucleotidase were increased by pilocarpine in hippocampus. Our results have indicated that previous treatment with AEDs was able to prevent the increase in hippocampal ecto-nucleotidases of pilocarpine-treated rats. These findings have shown that anticonvulsant drugs can modulate plastic events related to the increase of nucleotidase expression and activities in pilocarpine-treated rats. PMID- 17401673 TI - Assessment of antioxidant effect of 2,5-dihydroxybenzoic acid and vitamin a in brains of rats with induced hyperoxia. AB - The aim of this study was to evaluate the effect of 2,5-dihydroxybenzoic acid, a salicylate derived from Acetyl salicylic acid (ASA) and vitamin A (vit A) on Na(+), K(+) ATPase enzyme and GSH levels in brain of rats exposed to hyperoxia (Hyp) as oxidant protocol. Rats were treated as follow: group I (control), group II (Hyp), group III (Hyp, ASA), group IV (vit A), group V (Hyp, vit A), group VI (Hyp, vit A, ASA). Vit A was given 5 days before and during Hyp, aspirin at the end of Hyp. Na(+),K(+) ATPase and total ATPase activity was significantly increased in group V. Levels of GSH showed a significant increase in group III, besides, levels of 2,5-dihydroxybenzoic acid as salicylate in plasma were significantly increased in group II. These results elucidate differences in the biochemical response of animal towards intake of various types of antioxidant substances, with increased GSH and salicylate in hyperoxia. PMID- 17401674 TI - Thrombin causes the enrichment of rat brain primary cultures with ependymal cells via protease-activated receptor 1. AB - Ependymal cell culture models from rat have been developed over the last 20 years to facilitate biochemical studies on this least-studied glial cell type. The cell culture protocol calls for the presence of thrombin, which is essential for obtaining a high proportion of multiciliated ependymal cells. The serine protease appears to act via protease-activated receptor 1 to prevent the apoptosis of ependymal precursors and enhance their proliferation without affecting contaminating cells. Unciliated precursors differentiate into polyciliated ependymocytes by passing through a stage of monociliation. The message for protease-activated receptor (PAR) 1 is initially abundant in the cultures, but its level declines as the cells differentiate. Besides PAR 1, signalling through PAR 2 also promotes ciliation in rat brain primary cultures, albeit to a lesser degree than the thrombin receptor. Thrombin and other proteases may be involved in the regulation of ventricular wall development. This action would be mediated mainly by PAR1. PMID- 17401675 TI - A quantum-dot nanocrystal study of GABAA receptor subunits in living cerebellar granule cells in culture. AB - Quantum dots (QDs) are semiconductor nanocrystals emerging as a new class of fluorescent labels with large brightness, multi color fluorescence emission and resistance against photobleaching. Here we have used QDs as biological markers in an immunofluorescence approach. In this work GABA(A )receptors of rat cerebellar granule cells have been studied and in particular we have visualized the beta(2/3) and delta subunits in live cells. The results obtained were compared to those gathered with conventional probes. The images of the delta subunit in living cells appear to correspond to those expected for a subunit part of GABA(A )receptors mediating tonic inhibition in the granules cell bodies. PMID- 17401676 TI - PEN-2 overexpression induces gamma-secretase protein and its activity with amyloid beta-42 production. AB - PEN-2 is a component of the gamma-secretase complex, which is involved in the cleavage of the beta-amyloid precursor protein. The aim of this study was to determine the mechanism by which PEN-2 overexpression regulates gamma-secretase expression and the production of Abeta-42. In order to determine this, a hybrid gene harboring human PEN-2 was constructed, and used in the transfection of SK-N MC human neuroepitheliomal cells. This cell line was also co-transfected with a combination of human mutant presenilin 2 (hPS2m) and APPsw. Our results indicated that (i) human PEN-2 overexpression induced an increase in gamma-secretase activity and its proteins, including PS1-CTF, APH-1, and nicastrin, thus production of Abeta-42, (ii) co-transfection of human PEN-2 with both hPS2m and APPsw exerted no more profound effects on the induction of gamma-secretase proteins and its activity than did transfection with hPEN-2 alone. Thus, PEN-2 overexpression may facilitate assembly into the more active gamma-secretase complex, and may also induce an increase in activity, thus affecting Abeta-42 production. PMID- 17401677 TI - Phosphatidylinositol 4,5-bisphosphate induced flunitrazepam sensitive-GABAA receptor increase in synaptosomes from chick forebrain. AB - The flunitrazepam sensitive-GABA(A) receptor density was increased by cytochalasins C and D at 37 degrees C suggesting that microfilament depolymerization induces exposure to the radioligand of a GABA(A) receptor in synaptosomes (Pharm Biochem Behav 72 (2002) 497). Similarly, phosphatidylinositol 4,5-bisphosphate (1-5 microM), but not a mixture of phospholipids, induced an increase of GABA(A) receptors in synaptosomes. Furthermore, N-ethyl maleimide, an inactivator of the sensitive fusion protein, which interacts with GABA(A) receptor, abolished the receptor increase induced by phosphatidylinositol-4,5 bisphosphate. Together, the results suggest that phosphatidylinositol-4,5 bisphosphate, acts via microfilament depolymerization increasing the binding of the radioligand to receptors possibly by modulation of their interaction with proteins involved in trafficking and docking mechanisms. PMID- 17401678 TI - Changes in the spontaneous calcium oscillations for the development of the preconditioning-induced ischemic tolerance in neuron/astrocyte co-culture. AB - Spontaneous Ca(2+) oscillations are believed to contribute to the regulation of gene expression. Here we investigated whether and how the dynamics of Ca(2+) oscillations changed after sublethal preconditioning (PC) for PC-induced ischemic tolerance in neuron/astrocyte co-cultures. The frequency of spontaneous Ca(2+) oscillations significantly decreased between 4 and 8 h after the end of PC in both neurons and astrocytes. Treatment with 2-APB, an inhibitor of IP3 receptors, decreased the oscillatory frequency, induced ischemic tolerance and a down regulation of glutamate transporter GLT-1 contributing to the increase in the extracellular glutamate during ischemia. The expression of GLT-1 is known to be up-regulated by PACAP. Treatment with PACAP38 increased the oscillatory frequency, and antagonized both the PC-induced down-regulation of GLT-1 and ischemic tolerance. These results suggested that the PC suppressed the spontaneous Ca(2+) oscillations regulating the gene expressions of various proteins, especially of astrocytic GLT-1, for the development of the PC-induced ischemic tolerance. PMID- 17401679 TI - Effect of resveratrol on antioxidant enzyme activities in the brain of healthy rat. AB - We have studied the effect of resveratrol on lipoperoxidation and antioxidant enzyme activity level in the brain of healthy rats. When intraperitoneally administered, resveratrol significantly and dose dependently decreased brain malondialdehyde level. Resveratrol also increased in a dose-dependent way brain superoxide dismutase, catalase and peroxidase activities. Optimal effect on antioxidant enzyme and lipoperoxidation products were obtained with resveratrol concentration of 12.5 mg/kg body wt. Native polyacrylamide gel electrophoresis analysis of antioxidant isoenzymes revealed that resveratrol up regulated at least two acidic superoxide dismutase isoforms called A(1) and A(2), two basic isoforms called B(1) and B(2). Resveratrol also up regulated two catalase isoforms and a broad peroxidase band corresponding to several isoforms. All these findings suggest that resveratrol is able to cross the blood brain barrier and exerts potent antioxidant features. Resveratrol also exerts neuroprotective properties by up regulating several detoxifying enzymes, most of which are iron proteins. PMID- 17401680 TI - Triptolide upregulates NGF synthesis in rat astrocyte cultures. AB - Triptolide (T10), an extract from the traditional Chinese herb, Tripterygium wilfordii Hook F (TWHF), has been shown to attenuate the rotational behavior induced by D: -amphetamine and prevent the loss of dopaminergic neurons in the substantia nigra in rat models of Parkinson's disease. To examine if the neuroprotective effect is mediated by its stimulation of production of neurotrophic factors from astrocytes, we investigated the effect of T10 on synthesis and release of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) in rat astrocyte cultures. T10 did not affect the synthesis and release of either BDNF or GDNF. However, it significantly increased NGF mRNA expression. It also increased both intracellular NGF and NGF level in culture medium. These results indicate that the neuroprotective effect of T10 might be mediated, at least in part, via a stimulation of the production and release of NGF in astrocytes. PMID- 17401681 TI - Antioxidant effects of selegiline in oxidative stress induced by iron neonatal treatment in rats. AB - Increased levels of iron in specific brain regions have been reported in neurodegenerative disorders. It has been postulated that iron exerts its deleterious effects on the nervous system by inducing oxidative damage. In a previous study, we have shown that iron administered during a particular period of the neonatal life induces oxidative damage in brain regions in adult rats. The aim of the present study was to evaluate the possible protective effect of selegiline, a monoamino-oxidase B (MAO-B) inhibitor used in pharmacotherapy of Parkinson's disease, against iron-induced oxidative stress in the brain. Results have shown that selegiline (1.0 and 10.0 mg/kg), when administered early in life was able to protect the substantia nigra as well as the hippocampus against iron induced oxidative stress, without affecting striatum. When selegiline (10.0 mg/kg) was administered in the adult life to iron-treated rats, oxidative stress was reduced only in the substantia nigra. PMID- 17401682 TI - Effect of Berkson measurement error on parameter estimates in Cox regression models. AB - We study the effect of additive and multiplicative Berkson measurement error in Cox proportional hazard model. By plotting the true and the observed survivor function and the true and the observed hazard function dependent on the exposure one can get ideas about the effect of this type of error on the estimation of the slope parameter corresponding to the variable measured with error. As an example, we analyze the measurement error in the situation of the German Uranium Miners Cohort Study both with graphical methods and with a simulation study. We do not see a substantial bias in the presence of small measurement error and in the rare disease case. Even the effect of a Berkson measurement error with high variance, which is not unrealistic in our example, is a negligible attenuation of the observed effect. However, this effect is more pronounced for multiplicative measurement error. PMID- 17401683 TI - Bayesian analysis of generalized odds-rate hazards models for survival data. AB - In the analysis of censored survival data Cox proportional hazards model (1972) is extremely popular among the practitioners. However, in many real-life situations the proportionality of the hazard ratios does not seem to be an appropriate assumption. To overcome such a problem, we consider a class of nonproportional hazards models known as generalized odds-rate class of regression models. The class is general enough to include several commonly used models, such as proportional hazards model, proportional odds model, and accelerated life time model. The theoretical and computational properties of these models have been re examined. The propriety of the posterior has been established under some mild conditions. A simulation study is conducted and a detailed analysis of the data from a prostate cancer study is presented to further illustrate the proposed methodology. PMID- 17401684 TI - Community integration of adults with psychiatric disabilities and histories of homelessness. AB - This study tests components of Wong and Solomon's (2002, Mental Health Services Research, 4(2), 13-28) model of community integration, identifying both the dimensions and predictors of integration. It evaluates community integration among adults with psychiatric disabilities assigned randomly to receive either independent scatter-site apartments with the Housing First approach (experimental) or services as usual (control). Factor analysis supported a definition of community integration that includes psychological, physical, and social domains, but also suggested the existence of another factor, independence/self-actualization. Regression analysis suggested that choice and independent scatter-site housing were predictors of psychological and social integration respectively. Psychiatric hospitalization, symptomatology and participation in substance use treatment were also found to influence aspects of integration. We discuss several issues that future studies should explore including the possibility that the same factor can differentially influence discrete aspects of integration, the role of person-environment fit, integration that is not based in the neighborhood, and, finally, conceptions of community integration from the perspective of consumers themselves. PMID- 17401685 TI - Role of oxidative stress in celecoxib-induced renal damage in wistar rats. AB - Celecoxib, a selective cyclo-oxygenase-2 (Cox-2) inhibitor, prevents the formation of prostaglandins, responsible for maintenance of renal function. Celecoxib administration may lead to renal damage. Since free radicals and antioxidant mechanisms play a significant role in renal injury; this study was designed to evaluate the role of oxidative stress in celecoxib-induced renal damage. The administration of celecoxib resulted in moderate and mild tubulointerstitial nephritis in chronic and acute group. The renal function tests were significantly altered only in the chronic group. The results in both the acute and the chronic group showed (1) a significant increase in the lipid peroxidation and in the activities of superoxide dismutase, catalase and glutathione-S-transferase and (2) a decrease in nitrite, reactive thiols and glutathione. In conclusion, our study suggests that chronic administration of celecoxib may have a damaging effect on kidney, as evident through altered histopathology and renal functions. This damage may be mediated by oxidative stress. PMID- 17401686 TI - Long-term treatment with the combination of amantadine and ribavirin in hepatitis C nonresponders. A case series. AB - In this report, we describe five cases of chronic hepatitis C that have been treated with the combination of amantadine and ribavirin for an average of 44 months, emphasizing one case where the patient showed improvement in liver biopsy after treatment, worsening on removal, then a repeated improvement with re initiation. The five patients in this report belong to a pool of sixty patients from a 6 month pilot study using amantadine and ribavirin where treatment was subsequently continued. The mean ALT was 82.8+/-32 U/L pre-treatment and 33.8+/ 17.3 U/L post-treatment (p=0.02). The mean Knodell score was 7+/-1 pre-treatment and 3.6+/-1.5 post-treatment (p=0.13). The mean viral load was 584,155+/-248,027 lU/ml pre-treatment and 225,878+/-190,143 IU/ml post-treatment (p=0.05). In this case series we provide provocative data on the long-term use of ribavirin and amantadine in the HCV non-responder. PMID- 17401687 TI - Mucosal plug in the etiology of peptic ulcer disease. PMID- 17401688 TI - The role of smoking in Crohn's disease as defined by clinical variables. AB - The development of Crohn's disease (CD) is related to an interaction of genetic and environmental factors, with tobacco smoking being one of the most commonly studied environmental factors. In 1998, the Vienna classification was created to define CD by using three accepted clinical variables: age at diagnosis, disease behavior, and disease location. In this qualitative systematic review, articles examining the relationship between smoking and CD, using variables outlined in the Vienna classification, were identified utilizing multiple health databases. Current smoking was found to be associated with late-onset CD (> or =40 years old) and current smokers were more likely to progress to stricturing or penetrating type CD than were nonsmoking patients. Conflicting evidence exists regarding the relationship between smoking with respect to CD disease location. The Vienna classification is an important tool in permitting comparisons and predicting clinical course among CD cases, especially when smoking status is taken into account. PMID- 17401689 TI - Administrative issues in child psychiatry. AB - Administrative issues related to operating child and adolescent psychiatry programs or child mental health centers are substantially different than their adult counterpart programs. The increasing demands from managed care and other regulatory agencies make these programs difficult to operate. The smaller scale of these programs and the fewer existing programs make managing access to care more complicated. The administrators and clinicians in these programs have to be vigilant of legal responsibilities and reporting mandates that child practitioners and agencies that treat children need to abide by. In order to continue thriving, programs need to be efficient and fiscally viable. Issues such as building the continuum of care and finding the qualified personnel to staff these services are discussed in this article. PMID- 17401690 TI - Development of gradient descent adaptive algorithms to remove common mode artifact for improvement of cardiovascular signal quality. AB - BACKGROUND: Common-mode noise degrades cardiovascular signal quality and diminishes measurement accuracy. Filtering to remove noise components in the frequency domain often distorts the signal. METHOD: Two adaptive noise canceling (ANC) algorithms were tested to adjust weighted reference signals for optimal subtraction from a primary signal. Update of weight w was based upon the gradient term of the steepest descent equation: [see text], where the error epsilon is the difference between primary and weighted reference signals. nabla was estimated from Deltaepsilon(2) and Deltaw without using a variable Deltaw in the denominator which can cause instability. The Parallel Comparison (PC) algorithm computed Deltaepsilon(2) using fixed finite differences +/- Deltaw in parallel during each discrete time k. The ALOPEX algorithm computed Deltaepsilon(2)x Deltaw from time k to k + 1 to estimate nabla, with a random number added to account for Deltaepsilon(2) . Deltaw--> 0 near the optimal weighting. RESULTS: Using simulated data, both algorithms stably converged to the optimal weighting within 50-2000 discrete sample points k even with a SNR = 1:8 and weights which were initialized far from the optimal. Using a sharply pulsatile cardiac electrogram signal with added noise so that the SNR = 1:5, both algorithms exhibited stable convergence within 100 ms (100 sample points). Fourier spectral analysis revealed minimal distortion when comparing the signal without added noise to the ANC restored signal. CONCLUSIONS: ANC algorithms based upon difference calculations can rapidly and stably converge to the optimal weighting in simulated and real cardiovascular data. Signal quality is restored with minimal distortion, increasing the accuracy of biophysical measurement. PMID- 17401691 TI - Comparing perceived and objectively measured access to recreational facilities as predictors of physical activity in adolescent girls. AB - A number of studies in recent years have identified both self-report and objectively measured accessibility of recreational facilities as important predictors of physical activity in youth. Yet, few studies have: (1) examined the relationship between the number and proximity of objectively measured neighborhood physical activity facilities and respondents' perceptions and (2) compared objective and self-report measures as predictors of physical activity. This study uses data on 1,367 6th-grade girls who participated in the Trial of Activity for Adolescent Girls (TAAG) to explore these issues. Girls reported whether nine different types of recreational facilities were easily accessible. These facilities included basketball courts, golf courses, martial arts studios, playing fields, tracks, skating rinks, swimming pools, tennis courts, and dance/gymnastic clubs. Next, geographic information systems (GIS) were used to identify all the parks, schools, and commercial sites for physical activity located within a mile of each girl's home. These sites were then visited to inventory the types of facilities available. Girls wore accelerometers to measure their weekly minutes of non-school metabolic equivalent weighted moderate-to vigorous physical activity (MW-MVPA). The number of facilities within a half-mile of girls' homes strongly predicted the perception of easy access to seven out of nine facility types. Both individual facility perceptions and the total number of facilities perceived were associated with increased physical activity. For each additional facility perceived, girls clocked 3% more metabolic equivalent weighted moderate-to-vigorous physical activity (p < 0.001). Although girls tended to record 3% more of this kind of physical activity (p < 0.05) per basketball court within a mile of their homes, objective facility measures were otherwise unrelated to physical activity. The results from this study suggest that raising the profile of existing facilities may help increase physical activity among adolescent girls. PMID- 17401692 TI - Social capital and healthy urbanization in a globalized world. AB - This paper critically reviews the extent in which social capital can be a resource to promote health equity in urban contexts. It analyzes the concept of social capital and reviews evidence to link social capital to health outcomes and health equity, drawing on evidence from epidemiological studies and descriptive case studies from both developed and developing countries. The findings show that in certain environments social capital can be a key factor influencing health outcomes of technical interventions. Social capital can generate both the conditions necessary for mutual support and care and the mechanisms required for communities and groups to exert effective pressure to influence policy. The link between social capital and health is shown to operate through different pathways at different societal levels, but initiatives to strengthen social capital for health need to be part of a broader, holistic, social development process that also addresses upstream structural determinants of health. A clearer understanding is also needed of the complexity and dynamics of the social processes involved and their contribution to health equity and better health. The paper concludes with recommendations for policy and programming and identifies ten key elements needed to build social capital. PMID- 17401693 TI - Acute effects of different phosphorus sources on calcium and bone metabolism in young women: a whole-foods approach. AB - The recommended dietary phosphorus intake is exceeded in the typical Western diet. However, few studies have been conducted on the bioavailability and metabolic consequences of dietary phosphorus from different food sources. In this study, acute effects of dietary phosphorus from three different food sources and a phosphate supplement on calcium and bone metabolism were investigated. Sixteen healthy women aged 20-30 years were randomized to five controlled 24-hour study sessions, each subject serving as her own control. At the control session, calcium intake was ca. 250 mg and phosphorus intake ca. 500 mg. During the other four sessions, phosphorus intake was about 1,500 mg, 1,000 mg of which was obtained from meat, cheese, whole grains, or a phosphate supplement, respectively. The foods served were exactly the same during the phosphorus sessions and the control session; only phosphorus sources varied. Markers of calcium and bone metabolism were followed. Analysis of variance with repeated measures was used to compare the study sessions. Only the phosphate supplement increased serum parathyroid hormone (S-PTH) concentration compared with the control session (P = 0.031). Relative to the control session, meat increased markers of both bone formation (P = 0.045) and bone resorption (P = 0.049). Cheese decreased S-PTH (P = 0.0001) and bone resorption (P = 0.008). These data suggest that the metabolic response was different for different foods. PMID- 17401694 TI - Polyethylene particle-induced bone resorption in substance P-deficient mice. AB - Aseptic loosening is the major cause of total joint replacement failure. Substance P (SP) is a neurotransmitter richly distributed in sensory nerve fibers, bone, and bone-related tissue. The purpose of this study was to investigate the potential impact of SP on bone metabolism in polyethylene particle-induced osteolysis. We utilized the murine calvarial osteolysis model based on ultrahigh molecular weight polyethylene (UHMWPE) particles in 14 wild type mice (C57BL/J6) and 14 SP-deficient mice. Group 1 (C57BL/J 6) and group 3 (SP-knockout) received sham surgery, and group 2 (C57BL/J6) and group 4 (SP knockout) were treated with polyethylene particles. Analytical methods included three-dimensional micro-computed tomographic (micro-CT) analysis and histomorphometry. Bone resorption was measured within the midline suture. The number of osteoclasts was determined by counting the tartrate-resistant acid phosphatase-positive cells. UHMWPE-particle treated SP-deficient mice showed significantly reduced osteolysis compared to wild-type mice, as confirmed by histomorphometry (P < 0.001) and micro-CT (P = 0.035). Osteoclast numbers were significantly reduced in groups 3 and 4 compared to groups 1 and 2 (P < 0.001). Unexpectedly, SP-deficient mice (group 3) showed a significantly increased absolute bone mass compared to wild-type mice (group 1) (P = 0.02). The findings of our murine calvaria model lead to the assumption that SP is a promoter in particle-induced osteolysis. The pathophysiology of aseptic loosening is complex, and neuropeptides are not solely responsible for the progress of implant loosening; however, we conclude that there could be coherence between neurotransmitters and particle-induced osteolysis in patients with aseptic loosening. PMID- 17401695 TI - Bone formation zones in heterotopic ossifications: histologic findings and increased expression of bone morphogenetic protein 2 and transforming growth factors beta2 and beta3. AB - Heterotopic ossifications (HOs) formed after total endoprosthetic replacement of the hip joint were collected during revision surgery (n = 7). Tissues collected during regular hip arthroplasty (n = 12) were used as reference. Histomorphometric analysis was performed for assessment of bone formation activity in HOs and reference bone. HOs were dissected with histological guidance into three zones: formed bone, zone of active bone formation, and zone with fibrous connective and fibrocartilagineous tissue. Relative expression of the mRNA for bone morphogenetic protein 2 (BMP-2), transforming growth factor beta2 (TGF-beta2), and TGF-beta3 was determined by reverse-transcription polymerase chain reaction relative to beta-actin. Expression of all three growth factors was higher than in orthotopic bone. Similarly, the osteoid surface density was increased in HOs. The levels of all growth factors were higher in the zone of active bone formation or remodeling than in the zone of formed bone. In matured HOs, the osteoid surface density as well as mRNA levels were lower, although still significantly raised, indicating that bone formation slows down after 2 years. Immunohistochemical analysis demonstrated the presence of TGF-beta1, TGF beta2, TGF-beta3, and BMP-2 proteins in the zone of bone formation. We conclude that bone formation after heterotopic bone induction is initially intense, slows down within 2 years, and thereupon continues as active remodeling mainly on the border of HO. Our data indicate that BMP-2, TGF-beta2, and TGF-beta3 are involved in bone formation in HO. PMID- 17401697 TI - The health equity dimensions of urban food systems. AB - There is increasing recognition that the nutrition transition sweeping the world's cities is multifaceted. Urban food and nutrition systems are beginning to share similar features, including an increase in dietary diversity, a convergence toward "Western-style" diets rich in fat and refined carbohydrate and within country bifurcation of food supplies and dietary conventions. Unequal access to the available dietary diversity, calories, and gastronomically satisfying eating experience leads to nutritional inequalities and diet-related health inequities in rich and poor cities alike. Understanding the determinants of inequalities in food security and nutritional quality is a precondition for developing preventive policy responses. Finding common solutions to under- and overnutrition is required, the first step of which is poverty eradication through creating livelihood strategies. In many cities, thousands of positions of paid employment could be created through the establishment of sustainable and self-sufficient local food systems, including urban agriculture and food processing initiatives, food distribution centers, healthy food market services, and urban planning that provides for multiple modes of transport to food outlets. Greater engagement with the food supply may dispel many of the food anxieties affluent consumers are experiencing. PMID- 17401698 TI - [Dear female and male readers]. PMID- 17401696 TI - Analysis of the survival of H. pylori within a laboratory-based aquatic model system using molecular and classical techniques. AB - Despite the significance of Helicobacter pylori infection for man, its transmission is not clearly known. The human stomach is considered the reservoir of this pathogen, and one of the accepted routes is fecal-oral, in which water acts as a vector. However, although H. pylori epidemiology associates its transmission with water, only molecular and not cultural analysis detects the bacteria in water. This study was carried out to understand these data through studying the survival of H. pylori in a laboratory water model using cultural, morphological, and molecular methods. A mineral water system spiked with H. pylori and stored at 7 +/- 1 degrees C in the dark was analyzed by different methods over a period of 3 weeks. The total number of cells observed by DAPI staining and their DNA content remained constant over this study period. In contrast, cells could no longer be cultured after 5 days. Cell viability, which was determined via the LIVE/DEAD BacLight kit, decreased up to day 14, and at day 21 all cell membranes were damaged. In addition, a gradual conversion from spiral to coccal morphology occurred from day 3 onward. However, polymerase chain reaction (PCR) technique detected H. pylori DNA at day 21 and 3 months later. A study of the cell morphology of a young colony demonstrated the coexistence of bacilli and cocci. The results of this study show that H. pylori survives in water but loses its culturability and bacillar morphology rapidly, although it remains viable for longer periods and its DNA is still detectable much later. Thus, interpreting H. pylori's behavior in water differs according to the type of analysis. Consequently, we suggest that the presence of H. pylori infective cells is overestimated by PCR, whereas, in contrast, culture techniques underestimate it. Nevertheless, H. pylori should be considered a waterborne pathogen during its viable period, independently of its shape and culturability, as its presence in water may be risky for human health. PMID- 17401699 TI - [Importance of the history and the utilization of therapy guidelines in patients with arrhythmias]. AB - In this article criteria for taking the history in patients with manifest or potential life threatening arrhythmias are outlined. The importance of a sound (family) history is demonstrated. Besides specific correlates for arrhythmias in the history of a patient, the long term consequences of dyspnoe on exertion and hypertension are outlined. In 80% of patients sudden death is due to myocardial ischemia or interstitial myocardial fibrosis which in turn result from acquired diseases such as coronary artery disease and hypertension. Against this background the importance of the prevention of sudden cardiac death by nonantiarrhythmics drugs is stressed. Examples of under-utilization of therapy guidelines are given with special reference to beta-blocker therapy. PMID- 17401700 TI - [The surface ECG in the diagnosis of cardiac arrhythmias: the value of the right precordial leads]. AB - The surface electrocardiogram (ECG) is a simple noninvasive method to assess the electrical activity of the heart and provides important information to identify patients with cardiac arrhythmias and increased arrhythmic risk. This brief review highlights cardiac conditions in which the right precordial leads recorded on the surface ECG during sinus rhythm or tachycardia are of important diagnostic and prognostic value. Epsilon waves seen in the right precordial ST segments are the electrocardiographic hallmark of arrhythmogenic right ventricular cardiomyopathy. The diagnosis of Brugada syndrome and risk stratification of affected patients are based on a coved-type >or=2 mm ST-segment elevation in the right precordial leads. This typical ECG pattern may be present persistently, intermittently or only after administration of sodium-channel blockers. The early repolarization syndrome, most commonly seen in healthy young individuals, is characterized by a ST-segment elevation of 1 to 4 mm in the mid-precordial leads with a notched and elevated J point in lead V4. The precordial ECG T-wave repolarization pattern may be helpful in identifying the genotype in patients with suspected long QT syndrome. In patients with overt preexcitation, the surface leads V1 and V2 play a key role in localizing the site of bypass-tract insertion. Finally, the right precordial lead V1 is often crucial in the diagnosis of narrow and broad QRS-complex tachycardias. PMID- 17401701 TI - Non-invasive risk stratification: prognostic implications of exercise testing. AB - The interpretation of exercise ECGs is frequently limited to an assessment of the ST segment for evaluation of ischemia. However, exercise testing represents a simple test of the complex interactions of the autonomous nervous system and provides a multitude of information with independent prognostic information. This overview summarizes available studies on the significance of chronotropic incompetence, exercise capacity, heart rate recovery after exercise, and the incidence of ventricular premature activity during or after exercise. These parameters express subclinical pathophysiological changes which cannot be identified by other means, e. g., coronary angiography or echocardiography. A reduced maximal heart rate under exercise may indicate a "blunted" sympathetic reaction as a protective myocardial reaction to an increase in left ventricular wall stretch in preclinical forms of heart failure. A reduced exercise capacity, measured in metabolic equivalents, METs, is one of the strongest predictors of cardiac and all cause mortality. A reduced heart rate recovery after exercise indicates an increased sympathetic tone or lack physiological increase in vagal tone; increased ventricular ectopy potentially identifies preclinical cardiomyopathy. This summary of study results aims at providing a new view on exercise testing that should lead to an improved understanding of relevant information included in the exercise ECG. PMID- 17401702 TI - [Clinical significance of dynamic QT-interval-analyses]. AB - Dynamic parameters of ventricular repolarization as Holter derived parameters expressed as QT-interval adaptation to heart rate changes (QT/RR-slope) and QT interval-variability are being more and more frequently used to identify patients with increased risk for ventricular arrhythmias. Steep QT-RR-slopes, reflecting inadequate adaptation of ventricular repolarization to heart rate changes, as well as increased QT-interval-variability, reflecting temporal inhomogeneity of ventricular repolarization duration, are frequently observed in patients at risk for sudden cardiac death. Additionally, there is strong evidence for significant alterations in the dynamics of action potential duration restitution in patients with structural heart disease. This review gives an up-to-date overview about the current research in methods of assessment and clinical relevance of dynamic parameters of ventricular repolarization. PMID- 17401703 TI - Clinical significance of microvolt T-wave alternans. AB - Several studies have recently proven that primary preventive therapy of sudden arrhythmogenic death is possible in selected patients with congestive heart failure, particularly in the setting of ischemic cardiomyopathy [1, 2]. However, a number needed to treat between 11 and 17 to save one life over three years in these studies indicates that a more accurate identification of high risk patients is desirable in order to avoid unnecessary implants of cardioverter/defibrillators (ICD). Since currently available risk stratification methods have limited predictive accuracy, development of new techniques is important in order to non-invasively assess arrhythmogenic risk in patients prone to sudden death. Microvolt level T-wave alternans (mTWA) has recently been proposed to assess abnormalities in ventricular repolarization favoring the occurrence of reentrant arrhythmias [3, 4]. In 1994, a first clinical study by Rosenbaum and coworkers [5] convincingly demonstrated that mTWA is closely related to arrhythmia induction in the electrophysiology (EP) laboratory as well as to the occurrence of spontaneous ventricular tachyarrhythmias during follow-up [5]. More recently, a number of clinical studies has examined its clinical applicability [4-7]. The present review summarizes currently available clinical data on TWA with a particular focus on risk stratifying patients with congestive heart failure and myocardial infarction. PMID- 17401704 TI - [Progression of heart failure during cardiac resynchronization therapy: what is the cause?]. AB - A 74 year old patient with cardiac resynchronization therapy (CRT) since 12 months presents with cardiac decompensation and dyspnea NYHA III-IV after improvement of cardiac symptoms for 9 months. Her history is remarkable for left ventricular hypertrophy, aortic valve replacement 14 years earlier, a left ventricular ejection fraction of 35-40%, and permanent third degree AV block. The 12 lead standard ECG reveals the cause of cardiac deterioration which is confirmed by telemetry. This case demonstrates the importance of optimized programming of CRT systems which differs from conventional programming of dual chamber devices in some aspects. PMID- 17401706 TI - Diagnosing influenza--clinical assessment and/or rapid antigen testing? PMID- 17401707 TI - Non-neoformans cryptococcal infections: a systematic review. AB - Non-neoformans cryptococci have been generally regarded as saprophytes and rarely reported as human pathogens. However, the incidence of infection due to these organisms has increased over the past 40 years, with Cryptococcus laurentii and Cryptococcus albidus, together, responsible for 80% of reported cases. Conditions associated with impaired cell-mediated immunity are important risks for non neoformans cryptococcal infections and prior azole prophylaxis has been associated with antifungal resistance. The presence of invasive devices was a significant risk factor for Cryptococcus laurentii infection (adjusted OR = 8.7; 95% CI = 1.48-82.9; p = 0.003), while predictors for mortality included age > or =45 years (aOR = 8.4; 95% CI = 1.18-78.82; p = 0.004) and meningeal presentation (aOR = 7.0; 95% CI = 1.85-60.5; p= 0.04). Because clinical manifestations of non neoformans cryptococcal infections are most often indistinguishable from Cryptococcus neoformans, a high index of suspicion remains important to facilitate early diagnosis and prompt treatment for such infections. PMID- 17401708 TI - The role of Aeromonas in diarrhea: a review. AB - The evidence for an enteropathogenic role of Aeromonas spp. is still controversial. This review examines various parameters related to a causative role of Aeromonas and concludes that infraspecific subsets of strains with a particular array of enterotoxin genes are potential enteropathogens. The consequences for stool bacteriology are discussed. PMID- 17401709 TI - Lack of discriminating signs and symptoms in clinical diagnosis of influenza of patients admitted to the hospital. AB - OBJECTIVES: Rapid diagnosis of influenza in hospitalised patients is important to prevent the transmission of the infection in the hospital. This prospective observational cohort study was designed to determine the relationship between the clinical diagnosis of influenza made by the physician at admission and the presence of influenza virus in patients with respiratory tract infections. METHODS: This prospective observational cohort study was conducted in a large Dutch teaching hospital in a period of four weeks during the influenza season 2004/2005. All patients of 18 years and older, admitted with respiratory tract infections were included in the study. Clinical and laboratory parameters, chest radiograph (CR), blood and sputum cultures and nasopharyngeal swab for polymerase chain reaction (PCR) were obtained for each patient. In addition, the physicians opinion at admission whether this patient had influenza was recorded. RESULTS: A total of 78 patients were hospitalized with respiratory tract infections. In 41 (53%) of them influenza virus was detected by PCR. Among the patients that were positive for influenza virus by PCR, a clinical diagnosis of influenza was made in 18 cases (44%). Conversely, clinical diagnosis of influenza was made in 16 out of 37 patients in whom influenza virus was not detected by PCR. Neither C reactive protein, leucocytes count nor an infiltrate on CR were helpful in determining the cause of the respiratory tract infection. CONCLUSIONS: The present findings failed to demonstrate a significant relationship between the clinical diagnosis of influenza and PCR detection of the virus. Also, the virus was present at least twice more often than influenza was clinically diagnosed. As a consequence, the decision to take protective measures to control spread of the virus should not rely on the clinical diagnosis. PMID- 17401710 TI - Clinical predictors for death in HIV-positive and HIV-negative tuberculosis patients in Guinea-Bissau. AB - OBJECT: To assess easily monitored predictors for tuberculosis mortality. DESIGN: Risk factors for tuberculosis mortality were assessed during the 8-month treatment in 440 men and 269 women diagnosed with confirmed or presumed intrathoracic tuberculosis included prospectively in Guinea-Bissau from May 1996 to April 2001. A civil war occurred in the study area from June 1998 to May 1999. RESULTS: 12% were HIV-1 positive, 16% HIV-2 positive and 7% were HIV dually infected. Case fatality rates for HIV positive were higher during (35% [22/63]) and after the war (29% [27/92]) compared to before the war (17% [15/88]). The war did not have an effect on the case fatality rate in HIV negative (10% [13/135] before the war). HIV-1-infected patients had higher mortality than HIV-2 infected, mortality rate ratio (MRR) = 2.28 (95% confidence interval 1.17-4.46). Men had higher mortality than women but only among the HIV negative (MRR = 2.09 [0.95-4.59]). Hence, the negative impact of HIV infection on mortality was stronger in women (MRR = 6.51 [2.98-14.2]) than in men (MRR = 2.64 [1.67-4.17]) (test of homogeneity, p = 0.051). Anergy to tuberculin was associated with death in HIV positive (MRR = 2.77 [1.38-5.54]) but not in HIV negative (MRR = 1.14 [0.52-2.53]). Signs of immune deficiency, such as oral candida infection or leukoplakia (MRR = 4.25 [1.92-9.44]) and diarrhea (MRR = 2.15 [1.29-3.58] was associated with mortality in HIV positive. Tendencies were similar among HIV negative. HIV-positive relapse cases were at increased risk of dying (MRR = 2.42 [1.10-5.34]). Malnutrition, measured through mid-upper arm circumference (MUAC), increased the risk of death. CONCLUSION: Easily monitored predictors for mortality in tuberculosis patients include clinical signs of immune deficiency and low MUAC. PMID- 17401711 TI - Surveillance of genotypic resistance mutations in chronic HIV-1 treated individuals after completion of the National Access to Antiretroviral Program in Thailand. AB - BACKGROUND: Due to the establishment of the National Access to Antiretroviral Program for People who have AIDS (NAPHA), approximately 80,000 Thai HIV-1 infected patients received antiretroviral drugs through the NAPHA program, which was completed at the end of 2005. The development of drug resistance is required for access to ARV drugs. The objective of this study was to determine the prevalence of antiretroviral drug resistance in Thai HIV-1 treated individuals after completing the NAPHA program. PATIENTS AND METHODS: Viral genotypic resistance testing was carried out for 1,880 HIV-infected patients experiencing treatment failure, who enrolled during 2000-2005. All patients were in a follow up treatment with ARV drugs available in clinical practice. The genotype was performed with the TRUGENE HIV-1 kit to assess resistant mutations to reverse transcriptase inhibitors and to protease inhibitors. RESULTS: The frequency of ARV drug resistance has significantly increased after the National Access To Antiretroviral Program was implemented. The reverse transcriptase genes M184V/I (919/1,880; 48.9%) and K103S/H (416/1,880; 22.1%) were the most frequent in nucleoside reverse transcriptase and non-nucleoside reverse transcriptase, respectively. In the protease genes, minor mutations or polymorphisms were found in the majority. Thymidine analogue mutations were presented and increased over time. This study showed a sharp increase in the prevalence of mutations associated with the GPO-VIR combination; nevirapine (948/1,880; 50.4%), lamivudine (889/1,880; 47.3%), and stavudine (703/1,880; 37.4%) after the program was completed. CONCLUSION: With the increased availability of antiretroviral therapy in a resource-constrained country, antiretroviral drug resistance should be closely monitored. HIV-1 drug resistance testing to enable the salvage therapy will remain a priority in Thailand. Furthermore, resistance testing should also become routine before prescribing treatment, and the consequences of continuing to provide a failing regimen must be considered. PMID- 17401712 TI - Trichinella spiralis-outbreak in the Slovak Republic. AB - BACKGROUND: Trichinellosis is a zoonosis caused by nematode worms of the genus Trichinella and acquired through the consumption of raw or undercooked meat or meat products that harbour parasite larvae. The present report was designed to evaluate the course and circumstances surrounding the trichinellosis outbreak that occurred due to consumption of pork from a backyard pig in the southwest area of Slovakia in 2001. PATIENTS AND METHODS: Patients suspected of acquiring infection were serologically examined by ELISA, sera of patients with symptoms of clinical trichinellosis were additionally tested by Western Blot. In symptomatic patients haematological and biochemical parameters were assessed on day 45 p.i. RESULTS: An epidemiological investigation showed that the disease was linked to the consumption of infected pork and/or smoked pork products and affected the household members of four families. Out of 23 persons who had consumed the infected meat, 11 showed anti-Trichinella IgA, IgM and IgG antibodies in their sera by an ELISA against somatic and excretory-secretory antigens. Six of them developed clinical symptoms. Sera from symptomatic patients recognised bands of 64, 47, 45 and 43 kDa of crude T. spiralis antigens by Western blot. Using multiplex PCR analysis, parasite larvae isolated from pork were identified as T. spiralis, rarely occurring in Slovakia. CONCLUSION: The outbreak of human trichinellosis in an area where feral animals have been previously considered free of Trichinella was unexpected. Following the suspicion of trichinellosis being addressed, larvae were detected in meat and meat products from pig and the course of disease in patients was successfully controlled and managed. PMID- 17401713 TI - Hepatitis B virus infection in health care workers in Albania: a country still highly endemic for HBV infection. AB - BACKGROUND: Health care workers (HCW) have an elevated risk of acquiring and transmitting parenteral infections. The aim of this study was to evaluate the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) markers with the final goal to encourage HBV vaccination of the non-immune Albanian HCW. METHODS: Among 480 HCW enrolled, 92 were physicians, 246 were nurses/techniques, 120 were auxiliary workers and 22 were office workers. RESULTS: The HBsAg, anti HBc and anti-HCV prevalence were 8.1%, 70% and 0.6%, respectively. The highest (11.4%) HBsAg prevalence was observed in the youngest age group (20-30 years of age). High HBsAg prevalence (7.2-7.5%) was detected also in age groups above 30 years. The highest HBsAg prevalence (12.6%) was found in the auxiliaries. The anti-HBc prevalence increased significantly with age from 59% in HCWs younger than 39 years to 87% among those older than 50 years. After adjustments for different job categories, age older than 40 years remained independently associated with anti-HBc positivity (OR = 2.9; 95% CI 1.9-4.6) and inversely associated with the lack of HBV immunity or infection markers (OR = 0.4; 95% CI 0.2-0.7). Of 142 HBsAg negative and/or anti-HBc Ab negative sera, 28 (20%) tested positive for anti-HBs. The 114 remaining individuals with no HBV infection or immunity markers were vaccinated against HBV infection. CONCLUSIONS: A high HBV infection rate and low HBV vaccination coverage were found in Albanian HCW. Albania is a Mediterranean country still highly endemic for HBV infection and new strategies to promote HBV vaccination are to be adopted. PMID- 17401714 TI - Sensitivity of a whole-blood interferon-gamma assay among patients with pulmonary tuberculosis and variations in T-cell responses during anti-tuberculosis treatment. AB - BACKGROUND: Interferon-gamma (IFN-gamma) assays are new tests for tuberculosis (TB) infection, and T-cell responses may be correlated with antigen burden. However, it is unclear if IFN-gamma assays can be used to monitor response to TB treatment. MATERIALS AND METHODS: We measured T-cell responses to TB specific antigens in 60 Indian patients with microbiologically confirmed active pulmonary tuberculosis, before, during, and after TB treatment. Most patients were hospitalized and had moderate to advanced disease. IFN-gamma responses were measured using the commercial whole-blood Quanti-FERON-TB Gold In Tube (QFT-G) assay at three time-points: at diagnosis (N = 60), after 2 months of intensive treatment (N = 47), and at 6 months (treatment completion) (N = 39). RESULTS: At baseline, 44 of 60 (73%) patients were positive by QFT-G. At the second time point, 38 of 47 (81%) patients were positive. At treatment completion, 31 of 39 (79%) patients were positive. Changes in IFN-gamma responses over time were highly inconsistent--some individuals showed increases, while others showed decreases or no changes. Although the average IFN-gamma levels decreased slightly during treatment (not significant), the QFT-G sensitivity remained mostly unchanged during therapy. CONCLUSIONS: Our data suggest that the QFT-G assay has modest sensitivity in patients with moderate to advanced pulmonary disease, but our results do not show a clear correlation between antigen burden and T-cell responses. Further research is needed to understand the kinetics of T-cell responses during TB treatment. PMID- 17401715 TI - Mumps epidemic among young British citizens on the island of Crete. AB - BACKGROUND: Mumps is an infectious disease, occurring rarely in many countries including Greece, where approximately 50 cases are reported each year. In Crete, Greece's biggest island, an average number of four cases are reported annually. PATIENTS AND METHODS: The study population consists of all patients hospitalised because of mumps in our clinic. Clinical, laboratory and serological confirmation of the disease has been performed. RESULTS: During the tourist summer periods of 2004 and 2005, 39 patients with mumps have been hospitalized in our clinic in Crete. All patients but one were young tourists from Britain. The patients presented with parotid gland swelling and fever and some developed complications of the disease as well. CONCLUSION: The above findings are consistent with the mumps epidemic, now occurring in the UK. The disease is spreading among the Greek population as well, since six cases have been reported. This study investigates the reasons for this epidemic, focusing on the role of MMR vaccination. PMID- 17401716 TI - Chronic vasculitis and polyneuropathy due to infection with Bartonella henselae. AB - Bartonella henselae, the causative agent of cat scratch disease and bacillary angiomatosis, is associated with an expanding spectrum of diseases. Here, we report on a 40-year-old patient suffering from chronic recurrent painful ulcers of the toes, distal axonal sensomotor polyneuropathy and Raynaud's phenomenon. Biopsy of the sural nerve demonstrated an axonal neuropathy with a neurogenic muscular atrophy. Treatment with high dose corticosteroids had no beneficial effect. A biopsy taken from a recurring ulcer 7 years after the beginning of the disease revealed superficial ulcerated hyperkeratosis with subepithelial proliferation of small vessels compatible with a diagnosis of verruca peruana, however, without detection of microorganism. Serologic analysis revealed an elevated IFT titer of 1:1,024 against B. henselae. Treatment with erythromycin induced healing of the ulcer, remission of the vasculitis and the polyneuropathy, and a decline of the IFT titer. This case illustrates that B. henselae infection should be considered in patients with vasculitis and polyneuropathic syndromes. PMID- 17401717 TI - Seronegative Lyme neuroborreliosis in a patient on treatment for chronic lymphatic leukemia. AB - We report on a patient who developed seronegative Lyme neuroborreliosis complicating chemotherapy for chronic lymphatic leukemia. After the fifth cycle of chemotherapy (FCR: fludarabine, cyclophosphamide, rituximab and prednisone) the 63-year-old patient developed night sweat, arthralgia in elbows, wrists, proximal interphalangeal joints (PIPs) and strong neuropathic pain in both legs, followed by paresthesia and hypesthesia in the feet, arms and face. Laboratory analysis revealed an elevated C-reactive protein (CRP), a slight elevation of liver enzymes and decreased IgG levels. Cerebrospinal fluid (CSF) analysis showed a lymphomononuclear pleocytosis and an elevation of protein. A broad diagnostic work-up was negative including a negative Borrelia IgG and IgM ELISA. The patient did not remember recent tick bites, but after specific questioning he recollected a transient erythema on his leg developing just before the start of the last cycle of chemotherapy. As the combination of neuropathic pain and arthralgia, the transient erythema and the lymphomononuclear pleocytosis raised the suspicion of Lyme neuroborreliosis, the patient was treated for 3 weeks with ceftriaxone. On therapy all symptoms resolved and CRP normalized. Retrospective PCR analysis of a CSF sample confirmed the clinical diagnosis by detecting Borrelia garinii DNA. This case demonstrates that in immunosuppressed patients borrelial serology may be negative and that additional diagnostic approaches (including tests for direct Borrelia detection) may be needed to demonstrate borrelial infection. PMID- 17401718 TI - Conservative management of implantable cardioverter defibrillator-related endocarditis due to Bacillus spp. AB - Endocarditis is a devastating complication of implantable cardioverter defibrillator (ICD) therapy. Partial or complete device removal has been advocated for the treatment of this condition although controlled data are lacking. We present a case of ICD-related endocarditis caused by Bacillus spp. that occurred following coronary angiography. To the best of our knowledge, Bacillus spp. has not been previously described in such context. Moreover, conservative treatment with device retention was successful (no recurrence during a 6-year follow-up). Conservative management may be attempted in selected cases of ICD-related endocarditis, especially, those involving low-virulence organisms and rapid response to antibiotic therapy. This case also suggests that coronary angiography may be associated with transient bacteremia and subsequent infection of indwelling cardiac devices. PMID- 17401719 TI - Epstein-Barr virus infectious mononucleosis associated with acute acalculous cholecystitis. PMID- 17401720 TI - Repeated HCV-clearance in an HIV-infected patient. PMID- 17401721 TI - A two-step approach for screening contacts of active tuberculosis. PMID- 17401722 TI - In vivo Schild regression analyses using nonselective 5-HT2C receptor antagonists in a rat operant behavioral assay. AB - RATIONALE: Although competitive antagonism experiments are critical tools in the classification of potential pharmacotherapies, no studies have quantitatively compared the potencies of 5-HT(2C) receptor antagonists using the Schild regression analysis in vivo. OBJECTIVES: To evaluate the behavioral effects of 5 HT(2C) receptor agonists and antagonists, a series of nonselective 5-HT(2C) receptor antagonists, the 5-HT(2A/2C) receptor antagonist ketanserin, the 5 HT(2B) receptor antagonist SB 204,741, the 5-HT(2B/2C) receptor antagonist SB 200,646, and the peripherally acting 5-HT(2C) receptor antagonist RS102,221 were evaluated for their capacity to competitively antagonize the agonists MK212, mCPP, or BW723C86 in rats. MATERIALS AND METHODS: Male Sprague-Dawley rats (N = 28) were trained to respond under a fixed ratio 10 schedule of food reinforcement. A multiple-trial, cumulative-dosing procedure was used to evaluate the capacity of the compounds to suppress response rates. RESULTS: MK212, mCPP, and the 5-HT(2B) receptor agonist BW723C86 dose-dependently decreased response rates. Only metergoline, mianserin, and methysergide produced a dose-dependent antagonism of the rate-decreasing effects of both mCPP and MK212. Apparent pA(2) analysis indicated that metergoline, mianserin, and methysergide were approximately equipotent as antagonists overall. Metergoline and mianserin failed to block the rate-decreasing effects of BW723C86. Ketanserin, SB 200,646, SB 204,741, and RS102,221 failed to block either mCPP or MK212, suggesting that 5 HT(2A), 5-HT(2B), or peripheral 5-HT(2C) receptors do not play a primary role in the rate-decreasing effects of these two agonists. CONCLUSIONS: Taken together, these antagonism profiles suggest that the agonists MK212 and mCPP produce their rate-decreasing effects through a combination of 5-HT receptors with the 5-HT(2C) receptor playing a prominent but not exclusive role. PMID- 17401723 TI - Genotyping of Giardia duodenalis isolates among residents of slum area in Manila, Philippines. AB - Giardia duodenalis is a flagellated protist that causes gastrointestinal disease throughout the world. In the Philippines, study on G. duodenalis is limited. It is also believed that prevalence rates of this organism in the country are underestimated. In this study, stool samples from residents living in a slum area in Manila were collected. These were examined under microscopy for identification of common helminthic and protistan parasites. Results showed that 22.05% of 2,354 stool samples collected contained Giardia cysts. A fraction of samples (n = 133) positive for Giardia cysts were set aside. Genomic DNA was extracted from these samples and a polymerase chain reaction-restriction fragment length polymorphism procedure based on the organism's triose phosphate isomerase gene was utilized. This particular procedure is capable of distinguishing assemblages or genotypes within G. duodenalis. The highest identified assemblage was Assemblage B (86.47%). The two genotypes of Assemblage A were also detected. This is the first report on the identification of genotypes of G. duodenalis in the Philippines. The results of this study can serve as basis for future control and prevention of giardiasis and parasitism in the country. PMID- 17401724 TI - Correlation of appearance of metastasis-associated protein1 (Mta1) with spermatogenesis in developing mouse testis. AB - Mta1, a representative of the MTA gene family, is believed to be involved in the metastasis of malignant tumors. However, a systematic study of its physiological function has not been performed. It has been found in normal mouse organs at relatively low levels, except for in testis, suggesting a potential function in the male reproductive system. In order to explore the role of Mta1 protein during spermatogenesis, its expression in adult mouse testis was compared with that in developing mouse testis and in testis from adult mice treated with methoxyacetic acid, which selectively depletes primary spermatocytes. Quantitative analysis revealed that Mta1 protein gradually increased in the testis from 14 days postnatally. Immunolocalization analysis demonstrated strong signals in the seminiferous tubules, and Mta1 was predominantly present in the nucleus of primary spermatocytes and spermatogonia from 14 days postnatally. The most intensive staining was located in the nucleus of pachytene spermatocytes in mature testes. The expression pattern of Mta1 during spermatogenesis was also shown to be stage-specific by immunohistochemistry analysis. Finally, dramatic loss of Mta1 expression from pachytene spermatocytes was observed in the spermatogenic-arrested adult mouse testis. These results collectively demonstrate that Mta1 appears during postnatal testis development and suggest that this expression may be crucial for spermatogenesis. PMID- 17401725 TI - Neurogenesis and depression: what animal models tell us about the link. AB - There is growing evidence that stress causes a decrease of neurogenesis in the dentate gyrus and antidepressant treatment in turn stimulates the cell proliferation in the dentate gyrus. This has led to the hypothesis that a decreased neurogenesis might be linked to the pathophysiology of major depression. The article reviews the relationship of depressive-like behavior and neurogenesis in three animal models of depression with high validity: learned helplessness, chronic mild stress and chronic psychosocial stress of the tree shrew. All animal models provide evidence that stress which can lead to depressive-like behavior, in parallel causes a decrease of neurogenesis; vice versa, antidepressant treatment is able to revert not only behavioral changes but also to normalize neurogenesis. But the animal models argue against the notion that decreases of neurogenesis are the cause or the consequence of depressive like behavior since depressive-like behavior can occur without impairments in neurogenesis and decreasing neurogenesis does not neccessarily lead to depressive like behavior. This suggests that neurogenesis does not directly control affect but is tightly connected to the modulation of affect by stress and antidepressant measures. PMID- 17401726 TI - Adult hippocampal neurogenesis and aging. AB - The demographic changes in the foreseeable future stress the need for research on successful cognitive aging. Advancing age constitutes a primary risk factor for disease of the central nervous system most notably neurodegenerative disorders. The hippocampus is one of the brain regions that is prominently affected by neurodegeneration and functional decline even in what is still considered "normal aging". Plasticity is the basis for how the brain adapts to changes over time. The discovery of adult hippocampal neurogenesis has added a whole new dimension to research on structural plasticity in the adult and aging hippocampus. In this article, we briefly summarize and discuss recent findings on the regulation of adult neurogenesis with relevance to aging. Aging is an important co-variable for many regulatory mechanisms affecting adult neurogenesis but so far, only few studies have specifically addressed this interaction. We hypothesize that adult neurogenesis contributes to a neural reserve, i.e. the maintained potential for structural plasticity that allows compensation in situations of functional losses with aging. As such we propose that adult neurogenesis might contribute to the structural correlates of successful aging. PMID- 17401727 TI - Adult neurogenesis and the memories of drug addiction. AB - Evidence accumulated in the last decade indicating that psychoactive substances negatively influence neurogenesis in the adult hippocampus has provided new insights into the neurobiology of drug addiction. Using a variety of experimental approaches and treatments, drugs such as psychomotor stimulants, opioids, alcohol and psychedelic compounds have been shown to alter one or several aspects of adult neurogenesis, including the rate of progenitor proliferation, the survival of newly generated cells, and the maturation and acquisition of cellular phenotype. This evidence, though critical from a neurotoxicological standpoint, cannot be linked unambiguously to the process of drug dependence at this stage of research. Drug addiction is a complex recurrent process involving the acquisition and maintenance of drug taking, followed by detoxification, abstinence and eventual relapse to drug seeking. The specific contribution of adult hippocampal neurogenesis to each of these processes is yet to be determined. This notwithstanding, the suggested role of the hippocampus in the storage and retrieval of declarative and contextual memories on the one hand, and in the regulation of mood and affect on the other, provides a fertile ground for further exploring the mutual relationships between postnatal hippocampal neurogenesis and addictive behaviours. PMID- 17401728 TI - What causes the hippocampal volume decrease in depression? Are neurogenesis, glial changes and apoptosis implicated? AB - Even though in vivo imaging studies document significant reductions of hippocampal volume in depressed patients, the exact underlying cellular mechanisms are unclear. Since stressful life events are associated with an increased risk of developing depression, preclinical studies in which animals are exposed to chronic stress have been used to understand the hippocampal shrinkage in depressed patients. Based on morphometrical studies in these models, parameters like dendritic retraction, suppressed adult neurogenesis and neuronal death, all due to elevated levels of glucocorticoids, have been suggested as major causative factors in hippocampal shrinkage. However, histopathological studies examining hippocampi of depressed individuals have so far failed to confirm either a massive neuronal loss or a suppression of dentate neurogenesis, an event that is notably very rare in adult or elderly humans. In fact, many of the structural changes and the volume reduction appear to be reversible. Clearly, more histopathological studies are needed; especially ones that (a) employ stereological quantification, (b) focus on specific cellular elements and populations, and (c) are performed in nonmedicated depressed patients. We conclude that mainly other factors, like alterations in the somatodendritic, axonal, and synaptic components and putative glial changes are most likely to explain the hippocampal shrinkage in depression, while shifts in fluid balance or changes in the extracellular space cannot be excluded either. PMID- 17401729 TI - The association between anger-related personality trait and cardiac autonomic response abnormalities in elderly subjects. AB - Cardiac autonomic response abnormality associated with trait anger has been recognized to elevate blood pressure in daily life, leading to atherosclerotic progression and cardiovascular disease. To clarify the relationship between anger related personality traits and cardiac autonomic response in healthy elderly subjects, 54 volunteers consisting of 30 male (mean age 62.2+/-5.4) and 24 female (mean age 58.4+/-4.6) subjects underwent testing of heart rate variability (HRV) with head-up tilt. For the evaluation of trait anger, we used a questionnaire corresponding to the trait anger score taken from the State and Trait Anger Expression Inventory. Furthermore, we measured carotid intima-medial thickness (IMT) to evaluate atherosclerotic progression in subjects with anger trait. In female subjects, higher trait anger was positively associated with elevated carotid IMT and the suppression of HRV vagal attenuation from the supine to head up position, and negatively associated with the HRV sympathetic activity in the head-up position and also with the HRV sympathetic response from the supine to head-up position. In male subjects, trait anger was not significantly associated with carotid IMT or any HRV component with or without head-up tilt testing. We conclude that a simple noninvasive measure, short-term HRV with head-up tilt testing, could be a useful method to investigate the association between cardiac autonomic imbalance and increased risk of atherosclerosis associated with trait anger in healthy elderly subjects. PMID- 17401730 TI - Co-morbidity of adult attention-deficit/hyperactivity disorder with focus on personality traits and related disorders in a tertiary referral center. AB - OBJECTIVE: The prevalence and consequences of co-morbid axis-I and axis-II disorders as well as personality traits were examined in a large cohort of adult attention-deficit/hyperactivity disorder (AADHD) at a tertiary referral center. METHODS: In- and outpatients referred for diagnostic assessment of AADHD were screened. 372 affected probands were examined by means of the Structured Clinical Interview of DSM-IV axis-I/II disorders, the Revised NEO Personality Inventory (NEO-PI-R), and the Tridimensional Personality Questionnaire (TPQ). RESULTS: Lifetime co-morbidity with mood disorders was 57.3%, with anxiety disorders 27.2%, and with substance use disorders 45.0%. The histrionic personality disorder (35.2%) was the most frequent personality disorder. AADHD patients exhibited significantly altered scores on most of the NEO-PI-R and TPQ personality dimensions. The extent of substance abuse and dependence, as well as the presence of antisocial personality disorder alone or the cumulative number of other specific personality disorders was associated with lower psychosocial status (p<.0001). DISCUSSION: In a cohort of patients with AADHD referred to a single tertiary center co-morbidity with axis-I/II disorders was remarkably prevalent. In AADHD co-morbid mood, anxiety, and personality disorders as well as substance abuse/dependence is likely to be predictive of poor outcome. PMID- 17401731 TI - Predictors of engagement in the Alcoholics Anonymous group or to psychotherapy among Brazilian alcoholics : a six-month follow-up study. AB - OBJECTIVE: To ascertain factors associated with engagement of patients with alcohol dependence in Alcoholics Anonymous (AA) groups and psychotherapy. METHODS: About 300 hospitalized alcoholics were interviewed at hospitalization and again 3 and 6 months thereafter. Assessment included the administration of standardized instruments. Determinants of engagement in both interventions were assessed through logistic regression analysis. RESULTS: Higher educational level was predictive of engagement in AA after 6 months (OR = 2.19; CI 1.08-4.41). Engagement in psychotherapy after 6 months was related to having a university degree (OR = 3.60; CI 1.6-7.9), to a co-morbid depressive disorder (OR = 3.47; CI 1.8-6.5), to the use of other drugs together with alcohol (OR = 3.08; CI 1.5 6.19), to previous treatment (OR = 2.87; CI 1.29-6.40), and to having a high school degree (OR = 2.44; CI 1.24-4.80). The presence of substance-induced anxiety disorder was associated with non-engagement in psychotherapy (OR = 0.27; CI 0.63-0.003). CONCLUSION: The identification of predictors of engagement is important to guide clinicians in the choice of the treatment strategies that are more likely to be successful. PMID- 17401732 TI - A cross-sectional study to investigate current social adjustment of offspring of patients with schizophrenia. AB - OBJECTIVE: To investigate the impact of parental disorder in the life of adult offspring by evaluating education, current employment and marital status of a sample of offspring of patients with schizophrenia and comparing findings with population rates. METHODS: A sample of 489 patients with DSM-IV diagnoses of schizophrenia was identified in the public outpatient mental health services of the city of Cuiaba, Brazil. Of these patients, 294 had children, and a total of 828 offspring were identified. Data for 431 offspring aged 18 years or older were collected using a structured questionnaire answered by the patient-parent and a family member. RESULTS: The percentage of age-grade discrepancy for offspring aged 18 and 19 years was 59.2% (95%CI 45.4-73.0), not significantly different from the discrepancy rate for the same age group in the general population, which was 71.1%. Offspring of patients with schizophrenia had a significantly poorer employment situation than the general population (66.7% and 75.6%; 95%CI 62.1 71.3). Fewer male offspring were married than males in the general population (54.7% and 66.0%; 95%CI 48.2-61.2). CONCLUSION: Adult offspring of patients with schizophrenia had social adjustment problems that were markedly reflected in employment and marital status. PMID- 17401733 TI - "Add-On"-therapy with an individualized preparation consisting of free amino acids for patients with a major depression. AB - The efficacy of a deficit oriented add-on therapy with free amino acids in depressive patients treated with the antidepressant Remeron was evaluated. About 40 in-patients were investigated by a randomised double-blind placebo-controlled study during 4 weeks. Plasma levels of 20 amino acids and measures of depression, suicidal behaviour and aggression were surveyed on admission and after a 4 weeks' therapy with Remeron plus an individualized amino acid mixture or placebo. The preparation of the amino acid mixture was based on an aminogram and consisted of essential amino acids plus vitamins and trace elements as co-factors for the amino acid metabolism. Patients of the experimental group showed a significantly better improvement of depression and a higher responder rate than those of the placebo group. The results suggest that oral application of a deficit oriented amino acid mixture can improve the therapeutic outcome of an antidepressant. Furthermore, lacking side effects of the amino acids resulting also in a better patient compliance may improve the benefit/risk ratio. PMID- 17401734 TI - Revised diagnostic criteria for neuromyelitis optica (NMO). Application in a series of suspected patients. AB - We analyzed neuromyelitis optica (NMO) IgG in the serum or CSF samples from 46 patients with suspected NMO (28) and limited forms of NMO (18). One hundred and fifteen samples from multiple sclerosis (MS) patients were included as controls. The final clinical diagnosis after follow-up was 16 NMO, 12 MS, 11 transverse myelitis (TM) and seven recurrent optic neuritis (RON). NMO-IgG was detected in 62.5% of NMO, 50% of the recurrent longitudinally extensive TM, 14.3% of the RON but in none of the MS patients. The authors then compared the newly revised diagnostic criteria for NMO with the criteria published in 1999, in the 28 patients with suspected NMO. The revised criteria had higher specificity, and positive and negative predictive value (83.3% vs. 25%; 87.5% vs. 62.5; 83.3% vs. 75%), but slightly lower sensitivity (87.5% vs. 93.7%). Our study confirms NMO IgG as a highly specific marker of NMO, and the usefulness of the revised diagnostic criteria in predicting a diagnosis of NMO. PMID- 17401735 TI - Gait in attention deficit hyperactivity disorder : effects of methylphenidate and dual tasking. AB - BACKGROUND: Cognitive function and the loading of attention presumably play an important role in gait as well as in fall risk, but previous work has not demonstrated this in any cause-and-effect way. OBJECTIVES: To gain insight into the relationship between gait and cognitive function, we sought: (1) To compare the gait rhythmicity (stride time variability) of children with attention deficit hyperactivity disorder (ADHD) to controls, (2) To test the hypothesis that dual tasking leads to increased stride-to-stride variability in ADHD, and (3) To test whether pharmacological treatment that relieves ADHD symptoms reduces stride-to stride variability. PATIENTS AND METHODS: Gait was quantified in children with ADHD and in age-matched healthy controls under single task and dual task conditions on three occasions: off medications (both groups) and, in the ADHD group, after double blinded, randomized administration of methylphenidate (MPH) or placebo. RESULTS: At baseline, children with ADHD tended to walk with increased stride-to-stride variability compared to the controls during the single task condition (p = 0.09). During dual task walking, stride time variability was significantly reduced in the children with ADHD (p < 0.004), but not in the controls. In the children with ADHD, the placebo did not significantly affect stride-to-stride variability or the dual tasking response. In contrast, stride time variability was significantly reduced on MPH (p < 0.001) such that dual tasking no longer affected variability. CONCLUSIONS: The present findings demonstrate alterations in the gait of children with ADHD, support a cause and effect link between cognitive function and gait, and suggest that enhancement of attention abilities may, in certain populations, improve gait rhythmicity. PMID- 17401736 TI - Return of meningeal symptoms in a patient treated for cryptococcal meningitis. PMID- 17401737 TI - Postanoxic delayed-onset cerebellar syndrome. PMID- 17401738 TI - Does freezing in Parkinson's disease change limb coordination? A kinematic analysis. AB - The aim of this study was to analyse the kinematic characteristics of the strides before freezing and compare this with a voluntary stop and ongoing gait. Also, we investigated whether gait profiles were different as a function of the side of the body. Ten patients were included with a mean age of 64.8 years (SD 5.1). Within a Vicon 3D gait laboratory, patients performed several trials of normal walking and voluntary stops or were exposed to circumstances, which provoked freezing in the off-phase of the medication cycle. Spatiotemporal and key kinematic data of the four strides prior to freezing were compared between body sides and walking conditions using multiple regression models for repeated measures. Prior to freezing patients had severely decreased movement ranges in the sagittal plane (ranging between 31% and 61.5%), most notably in the ankle and hip joints. The general shape of movement remained in the pre-freezing profiles with largely intact dissociation of knee and hip movement in stance but reduced dissociation in swing. Also present were reduced push-off movements in the ankle with fixed dorsiflexion, increased flexion in hip and knee and anterior tilt of the pelvis. During both voluntary (stopping) and involuntary deceleration (freezing), the body side with the last complete stride before the freeze, showed significantly smaller joint ranges (p < .01). Body side differences were larger than sequential deterioration of consecutive steps within each side. Freezing is distinct from normal deceleration of gait in that the reduction of propulsive movement is much greater. Despite hastening of steps, timing deficits did not affect overall movement shapes, except for the tendency to have a flexed walking pattern. The side of the body where gait terminated before freezing was in most cases the side of symptom-dominance, but not consistently so. PMID- 17401739 TI - Spinal arteriovenous shunts presenting as intracranial subarachnoid haemorrhage. AB - BACKGROUND: In approximately 5% of patients with intracranial subarachnoid haemorrhage (SAH), the cause is another than a ruptured aneurysm or perimesencephalic haemorrhage. One of these causes is a spinal arteriovenous shunt (SAVS). The aim of this study was to investigate the characteristics of patients with SAVS who present with intracranial SAH without symptoms and signs suggesting a spinal cause. METHODS: We systematically reviewed the literature and searched the SAH database of the University Medical Center Utrecht, The Netherlands, for patients with SAVS presenting with intracranial SAH and studied the characteristics of patients with SAVS whose clinical presentation mimicked intracranial SAH caused by rupture of a saccular aneurysm. RESULTS: Thirty-five patients were identified after a review of the literature. In our SAH database, comprising 2142 patients included in the period 1985-2004, we found one patient (0.05%, 95 % CI 0.006- 0.3%). SAH due to SAVS occurred at any age (4-72 years). The SAVS was located at the craniocervical junction in 14 patients, at the cervical level in 11, and at the thoracolumbar level in the remaining 11 patients. The majority of patients (n = 26, 72%) had no disabling deficits at discharge or follow-up. CONCLUSION: Rupture of a SAVS presenting as intracranial SAH is rare and can occur at any age. The SAVS can be located not only at the craniocervical junction or cervical level but also in the thoracolumbar region. Most patients with SAVS presenting as intracranial SAH have a good recovery. PMID- 17401740 TI - Pioneers in neurology. Edward Flatau (1868-1932). PMID- 17401741 TI - Isolated paraneoplastic optic neuropathy associated with small-cell lung cancer and anti-CV2 antibodies. PMID- 17401742 TI - Spread of primary dystonia in relation to initially affected region. AB - Not only childhood-onset, but also adult-onset primary dystonia may spread to multiple body parts. The relative risk of spread by site of onset of dystonia, important for clinical prognosis and approach, has not been well characterized. The aim of this study was to prospectively follow the spread of dystonia in 132 consecutive patients and to estimate the risk of spread by the site of onset of dystonia. The patients were included in the study if primary focal dystonia was the only sign of neurological disease other than tremor; i.e. in all patients a single body part could be identified as affected at the onset. At the end of the followup (mean duration 7.5 years; range 5.2-13.4 years), 96 patients (73%) remained focal, while 26 (20%) and 10 (7%) progressed to segmental and generalized dystonia, respectively. The highest likelihood for further spread was observed in patients with initial blepharospasm (10 out of 30 patients; 33.3%), followed by dystonia of upper extremities (32.3%), torticollis (19.6%), and laryngeal dystonia (6.7%). In addition to the highest risk for further spread of dystonia, blepharospasm was associated with the fastest rate of spread (the second region affected on average after 1.2 years). Our results demonstrated that the initial site of primary dystonia was relevant for the risk of spread. PMID- 17401743 TI - Mal de debarquement: pseudo-hallucinations from vestibular memory? PMID- 17401744 TI - Gadolinium encephalopathy due to accidental intrathecal administration of gadopentetate dimeglumine. PMID- 17401745 TI - The effect of levetiracetam in startle disease. PMID- 17401746 TI - High concurrent presence of disability in multiple sclerosis. Associations with perceived health. AB - OBJECTIVES: (1) To explore functioning and concurrent presence of disabilities - concerning cognition, manual dexterity, walking, energy, mood, activities of daily living (ADL), and social/lifestyle activities - in persons with multiple sclerosis (PwMS) cared for at an outpatient MS clinic. 2) To describe the PwMS' perceived physical and psychological impact and associations with the same disabilities. MATERIAL/METHODS: A descriptive cross-sectional study was carried out in 219 PwMS at the MS Centre, Karolinska University Hospital. Logistic regression employing proportional odds models was used to identify the associations of the disabilities with the perceived physical and psychological impact. RESULTS: In this sample the distribution with regard to disease severity as per Expanded Disability Status Scale was; mild 59.5%, moderate 17% and severe 23.5%. Despite the high proportion with mild disease severity disability regarding cognition was found in 49%, manual dexterity 76%, walking 43%, energy 67%, mood 29%, ADL 44% and social/lifestyle activities in 48%. Two or more disabilities were found in 80%, 24 % had six or seven disabilities. Disability regarding energy, mood, walking, manual dexterity and ADL was significantly associated with increase in the perceived physical impact, whereas disability in energy and mood was significantly associated with increase in the perceived psychological impact. CONCLUSIONS: The presence of several concurrent disabilities, some significantly associated with high perceived physical and psychological impact, in the majority of PwMS in outpatient clinics highlights the importance to identify disabilities, in particular fatigue and depressed mood, in order to supply health care interventions aiming to improve the life situation of PwMS. PMID- 17401747 TI - Mechanism of multiple infarcts in multiple cerebral circulations on diffusion weighted imaging. AB - Acute multiple infarcts in multiple cerebral circulations (AMIMC) are thought to suggest the presence of cardioembolic sources or systemic hypercoagulopathy. However, the mechanism and the simultaneous occurrence of AMIMC are not well known. We reviewed 685 consecutive acute ischemic stroke patients who underwent diffusion-weighted imaging (DWI) within 48 hours of onset. AMIMC was defined as multiple acute DWI lesions distributed in more than one cerebral circulation (i.e., 2 anterior and 1 posterior circulations). Signal intensities on apparent diffusion coefficient (ADC) maps corresponding to acute DWI lesions were classified as 'low', 'iso-' or 'high' signals. Blood markers obtained within 24 hours after admission were compared between patients with and without AMIMC. Sixty-seven (9.8%) patients had AMIMC. Frequency of cardioembolism in AMIMC patients was only 29.9% (20/67), which was not different from non-AMIMC patients (21.7%, p = 0.16). Large-artery atherosclerosis (LAA) or small-vessel occlusion (SVO) in multiple circulations or combined LAA and SVO were identified in 34.3% (23/ 67) of AMIMC patients, although pure LAA and pure SVO were less frequent than in non-AMIMC patients. ADC signals were purely 'low' in 38 (56.7%) and 'mixed' (low with iso- or high) in 29 (43.3%). Cardioembolism tended to be associated with 'low' ADC signals (75.0%) compared with other stroke mechanisms (48.9%; p = 0.062). C-reactive protein was higher in AMIMC than in non- AMIMC patients (p = 0.009). Stroke mechanisms responsible for AMIMC are heterogeneous. ADC findings suggest that AMIMC commonly occur stepwise and may be useful in determining stroke mechanism. Systemic inflammation may be associated with the pathogenesis of AMIMC. PMID- 17401748 TI - Immunoglobulin-responsive dysautonomia in Sjogren's syndrome. PMID- 17401749 TI - Transcriptional profiling by cDNA-AFLP reveals novel insights during methyl jasmonate, wounding and insect attack in Brassica napus. AB - Plants exploit a broad range of defense mechanisms to effectively combat invasion by pathogens or herbivores. Each environmental stress activates multiple signal transduction pathways to ensure an effective spatial and temporal defense response. A detailed transcriptome analysis using the cDNA-AFLP technique was performed to identify genes that are differentially expressed in oilseed rape (Brassica napus cv. Westar) leaves upon treatment with methyl jasmonate, mechanical wounding, or feeding by diamondback moth larvae (Plutella xylostella). In total, 16 different primer combinations were used, generating cDNA fragments ranging from 50 bp to 500 bp in size. This technique generated an average of 60 amplification products per reaction and therefore a total number of 5,600 fragments per treatment. Out of 16,800 bands, 124 showed qualitative differences among the treated and their respective control samples, including 95 up-regulated and 29 down-regulated bands. Expression of a selected subset of differentially expressed genes was confirmed by Northern blot analysis. Sequencing of fragments grouped many of the expressed genes in the categories of signaling and wound or pathogen response with examples like Jacalin, Strictosidine synthase and MD-2-LPS homologs. Genes with altered expression in distal tissue included those involved in cellular housekeeping functions, suggesting modified resource allocation needed to respond to different stress conditions. Differences in local and systemic response as well as among the three different challenges were observed. Several new transcripts were identified that may play a role in insect attack and other signal transduction pathways. PMID- 17401751 TI - [Diagnostic value of tissue Doppler parameters in the early diagnosis of cardiomyopathies]. AB - Tissue Doppler and its derived parameters strain and strain rate show characteristic changes in patients with cardiomyopathy. Longitudinal systolic (S) and early diastolic (E') peak tissue velocities are reduced in the basal segments of the left ventricle in patients with dilated, hypertrophic, and restrictive cardiomyopathies, and in the right ventricular free wall in patients with arrhythmogenic dysplasia of the right ventricle. Similarly, strain and strain rate are lower than in normals. These changes are detectable in asymptomatic, genetically affected patients and in early stages of cardiomyopathy, where conventional echo signs such as impaired left ventricular ejection fraction or increased wall thickness are not yet present. Thus, tissue Doppler analysis may contribute to better identifying carriers of disease and subclinical early stages of cardiomyopathy. However, lack of measurement standardization and small numbers of cardiomyopathy patients examined by Doppler, together with difficulty in defining reliable normalcy ranges, still hamper widespread clinical use of this new tool. PMID- 17401752 TI - Current role of cardiac computed tomography. AB - The introduction of recent generations of multidetector computed tomography scanners has brought about substantial improvements in spatial and especially temporal resolution which have made imaging of the heart and, under certain conditions, visualization of the coronary arteries possible. Non-enhanced scans allow visualization of cardiac and coronary artery calcification. After intravenous injection of contrast agent, it is possible to visualize cardiac chambers and to analyze the coronary artery lumen. Morphological imaging of the heart, for example in the context of congenital heart disease, is possible by computed tomography (CT) and can, in some very selected cases, be clinically useful as an adjunct to echocardiography or magnetic resonance imaging. Functional imaging allows to analyze left and right ventricular function by CT.CT plays a more prominent clinical role in the context of coronary artery visualization. Coronary calcifications are indicative of coronary atherosclerosis and the presence and amount of coronary artery calcium have a strong predictive value concerning future cardiovascular events, even in asymptomatic individuals. It can potentially be used for refined risk stratification. Contrast-enhanced "CT coronary angiography" has become quite reliable under certain conditions (e.g., a low heart rate). The negative predictive value to rule out coronary artery stenoses has been found to be high in several studies. Therefore, clinical utility to rule out significant coronary artery stenoses in patients who are symptomatic, but have a low to intermediate pretest likelihood of disease is assumed. Ideally, "negative" coronary angiograms would be avoided by using coronary CT angiography. Large-scale trials that would pinpoint specific patient groups to benefit from "CT coronary angiography" are currently lacking. PMID- 17401753 TI - [Assessment of the natural history of coronary artery calcification and identification of its determinants. Rationale of the 2nd part of the Heinz Nixdorf Recall Study]. AB - The Heinz Nixdorf Recall Study, which was inaugurated in 2000, is an ongoing population-based study to evaluate the prediction of cardiovascular events by integrating new imaging and nonimaging modalities in risk assessment. A focus is the additional prognostic value of coronary artery calcification (CAC). Currently used risk stratification algorithms often describe the individuals' risk based on few established risk factors only inaccurately. Using noninvasive quantification of CAC progression, the natural history of atherosclerosis with its repetitive, frequently subclinical plaque ruptures, may detect an unstable course of the disease long before the disease irreversibly manifests in sudden death or myocardial infarction. While the independent additional prognostic value of CAC quantification has been shown in asymptomatic patients at intermediate risk, only few studies provided evidence for an independent prognostic value of serial CAC measurements. In the Heinz Nixdorf Recall Study, the impact of established and new risk factors, e.g., the metabolic syndrome, psychosocial and environmental risk factors, or genetic variables, can be assessed. Further, the association of CAC progression with the incidence of other cardiovascular diseases such as heart failure or aortic or aortic valve calcification can be described. Since April 2006, the participants of the study return to the study center 5 years after baseline recruitment to assess health status and to determine the risk factor profile. Based on recently published data, serial CAC measurements have been granted allowing for (1) characterization of the natural history of CAC progression, and (2) identification of its determinants. The rationale of serial CAC quantification is discussed in this article.The Heinz Nixdorf Recall Study will contribute to the appraisal of new imaging modalities in risk stratification. PMID- 17401754 TI - [Correlative imaging in cardiology. Principles and clinical application]. AB - Technical progress has engendered a broad spectrum of methods to image cardiac structure, function, and metabolism. Frequently, the imaging tools available provide complementary information. Therefore, methods to integrate image information from different modalities into one common coordinate system are increasingly receiving attention also in cardiology. Currently available technology includes software-based image fusion as well as hardware-based registration of datasets. The latter capitalizes on so-called hybrid cameras combining detectors of different modalities in one gantry. Hardware-based image fusion is, to date, anatomically more accurate than the software-based methodology. However, the anatomic accuracy of both approaches is still far from perfect. This is, in particular, due to artifacts caused by respiratory movements also affecting the heart. The clinical potential of correlative imaging of the heart includes an improvement of accuracy in diagnosing hemodynamically significant coronary artery disease using single-photon emission computed tomography (SPECT). This is due to the possibility to correct myocardial scintigraphy for attenuation artifacts using registered X-ray computerized tomographic (CT) images. The visualization of coronary anatomy and myocardial perfusion in one imaging session using hybrid systems combining CT with positron emission tomography (PET) or SPECT is also an interesting option. Nevertheless, larger clinical trials investigating its usefulness are still missing. The possibility to match structure with radioactivity concentration is essential for approaches to image the molecular composition of atherosclerotic plaques and their stability. PMID- 17401755 TI - Significance of late gadolinium enhancement in cardiovascular magnetic resonance imaging (CMR). AB - Cardiovascular magnetic resonance imaging (CMR) permits optimal differentiation between normal and diseased myocardium with the use of gadoliniumbased contrast agents and special magnetic resonance pulse sequences. Imaging is performed 10-20 min after contrast agent application to produce so-called late gadolinium enhancement (LGE) images which depict diseased myocardium with excellent reproducibility. Areas showing LGE correspond to zones of myocyte necrosis or myocardial fibrosis as shown by comparison with histopathology. Typical patterns of hyperenhancement exist in ischemic heart disease but also in dilated cardiomyopathy, hypertrophic cardiomyopathy and other inflammatory or infiltrative myocardial disease and are described in this article. LGE-CMR is helpful to distinguish advanced ischemic heart disease from nonischemic dilated cardiomyopathy. In ischemic heart disease LGE can also predict the functional recovery after revascularization procedures by directly showing the remaining viable myocardium. LGE may also become useful to predict malignant arrhythmias in patients with ischemic heart disease or nonischemic cardiomyopathy. This may lead in future to an increased role of LGE-CMR as a prognostic tool. PMID- 17401756 TI - [Evidence-based application of cardiac magnetic resonance and cardiac computed tomography for primary diagnosis of stable coronary artery disease with special attention to disease management programs and the German National Medical Care Guidelines]. AB - BACKGROUND AND PURPOSE: Despite all the progress made in diagnosis and treatment, cardiovascular diseases remain the no. 1 cause of death worldwide. In 2005, 27.9% of males and 24.1% of females (eight times more than for mammary carcinoma) in Germany died of coronary artery disease (CAD). Although mortality rates following acute myocardial infarction have considerably dropped, morbidity has increased- people are getting older, but they are getting older sick. The resulting need for reduction of CAD morbidity can only be achieved by truly early detection of patients at high coronary risk before occurrence of a coronary event. Modern imaging techniques like cardiac magnetic resonance (MR) and cardiac computed tomography (CT) are being increasingly utilized. The goal of this review is the practical application of evidence-based recommendations by relevantly and cost effectively implementing cardiac MR and cardiac CT with special attention to current national and international guidelines and recommendations. THE PATIENT WITH STABLE CHEST PAIN: The primary objective here is to ascertain if the described symptoms can be attributed to a stenosing CAD with inducible myocardial ischemia or if they are effected by extracardial sources. As always, patients' history, examination findings and the stress ECG play the major roles. The conventional approach prescribes that an abnormal resting ECG compromising the interpretation of a stress ECG, should immediately be followed by an imaging ischemia diagnosis technique, like stress echocardiography, myocardial scintigraphy or cardiac MR (recommendation I B). This also holds true when a stress ECG is assessable and the probability of a stenosing CAD is between 10% and 90% (recommendation I B or "appropriate"). Alternatively, "modern" procedures allow imaging ischemia diagnosis to be replaced by noninvasive coronary angiography using a CTA (recommendation IIa B). If the image quality by CTA is assessable (free of artifacts and no disruptive calcified plaques) and no coronary stenosis can be found, the coronary diagnosis process may at this point be terminated, avoiding superfluous cardiac catheterizations. If the CTA shows the possibility of a higher-grade coronary stenosis, a cardiac catheterization examination with stand-by PCI (percutaneous coronary intervention) can be performed. THE ASYMPTOMATIC PREVENTION PATIENT: The prim ary objective in cardiovascular primary prevention is the avoidance of a first myocardial infarction and/or a first stroke. Current guidelines for prevention of cardiovascular diseases recommend administration of acetylsalicylic acid (ASA) and a statin when risk>20%/10 years (recommendation I A). The coronary calcium score has a strong predictive power which is independent of conventional risk factors and thus offers the most relevant information in addition to Framingham, PROCAM or ESC scores regarding coronary risk. For patients initially showing "intermediate" coronary risk (10-20%), guidelines suggest the determination of the coronary calcium score, which leads to better risk assessment and to identification of patients needing more aggressive lipid lowering (recommendation IIb B). Thus, the cardiac CT, on the one hand, aims the intense risk reduction needed in primary prevention to the high-risk patients, on the other hand, it avoids "superfluous" cardiac catheterizations, unnecessary statin therapies and potentially harmful ASA administrations. However, the proof of coronary calcium must not be confused with the presence of coronary artery stenoses: a positive calcium score in an asymptomatic person does not by itself indicate the need for a cardiac catheterization. PMID- 17401757 TI - [Diagnostic and therapeutic measures in leaftlet escape of mechanical heart valves]. AB - Today, a variety of mechanical and biological heart valve prostheses is available for the treatment of heart valve failures. Leaflet escape is a rare but serious complication of heart valve replacement therapy with mechanical heart valves. Since the risk of leaflet escape is extremely rare compared to other postoperative complications, there is usually no need for a prophylactic reoperation. However, if the event is misdiagnosed, leaflet escape can be life threatening. Each cardiologist should be knowledgeable about the correct actions to take in the event of a leaflet escape. The patient should also be aware of the signs of leaflet escape and report any suspicious symptoms immediately. PMID- 17401758 TI - Familial hypertrophic cardiomyopathy. PMID- 17401759 TI - Exclusion of atherosclerotic plaque from the circulation using stent-grafts: alternative to carotid stenting with a protection device? AB - PURPOSE: To retrospectively assess the feasibility, safety, and clinical mid-term outcome of patients undergoing carotid artery stenting with stent-grafts. METHODS: Over a 4 year period stent-grafts were used in the endovascular treatment of symptomatic internal carotid artery stenosis in 12 patients (2 women, 10 men, aged 47-83 (mean 64) years). Protection devices were not used. Possible microembolic complications were evaluated by magnetic resonance imaging (MRI) examinations of the brain before and the day after the procedure in all patients. Mean follow-up was 22 months (range 1-42 months), by Doppler ultrasonography and conventional angiography as well as clinical examination. RESULTS: The technical success rate was 100%. A total of 13 coronary stent-grafts were used. The mean stenosis rate (in terms of diameter) was 85% and the mean length of stent-grafts used was 20.9 mm. The mean diameter to which the stent grafts were dilated was 4.66 mm. In-hospital complications occurred in 1 patient who suffered a minor femoral access hematoma that did not require transfusion or surgical decompression. Post-stenting diffusion-weighted MRI revealed several ipsilateral silent microemboli in only 1 case, which was completely asymptomatic. Two patients had a major stroke after 2 years of follow-up. Restenosis was found in 2 patients who underwent successful balloon dilatation followed by placement of a self-expandable bare stent within the stent-grafts. CONCLUSIONS: Stent grafts may prevent microembolic complications during stenting of atherosclerotic carotid lesions in selected cases, offering immediate exclusion of the atherosclerotic lesion from the circulation by pressing the plaque against the vessel wall. Comparative, randomized studies in larger series of patients are needed with carotid-dedicated stent-graft designs. PMID- 17401760 TI - Pulmonary arterial stent implantation in an adult with Williams syndrome. AB - We report a 38-year-old patient who presented with pulmonary hypertension and right ventricular dysfunction due to pulmonary artery stenoses as a manifestation of Williams syndrome, mimicking chronic thromboembolic pulmonary hypertension. The patient was treated with balloon angioplasty and stent implantation. Short term follow-up showed a good clinical result with excellent patency of the stents but early restenosis of the segments in which only balloon angioplasty was performed. These stenoses were subsequently also treated successfully by stent implantation. Stent patency was observed 3 years after the first procedure. PMID- 17401761 TI - Visceral and renal artery complications of suprarenal fixation during endovascular aneurysm repair. AB - BACKGROUND: The effect of suprarenal fixation of endovascular grafts on renal and visceral artery function remains undefined. This study aimed to determine renal and visceral artery complications following suprarenal fixation during endovascular aneurysm repair (EVR). METHODS: Prospectively collected data from 112 patients who received suprarenal fixation (group SF) and 36 patients who received infrarenal fixation (group IF) in a single institution from December 1997 to April 2005 were reviewed retrospectively. Median follow-up was 26 months (range 0.1-101 months). RESULTS: Stent struts extended to or above the level of 106 (94.6%) right renal arteries, 104 (92.9%) left renal arteries, 49 (43.8%) superior mesenteric arteries (SMA), and 7 (6.3%) celiac arteries in group SF. This group had 2 (1.8%) unintentional main renal artery occlusions, of which 1 was successfully treated at the first procedure with a renal stent. There was 1 (0.9%) SMA occlusion which resulted in bowel infarction and death. Group IF had no renal or visceral artery complications. There were no late-onset occlusions or infarcts. There was no significant difference in median serum creatinine between groups SF and IF at 1 month (p = 0.18) and 6 months to 12 months (p = 0.22) follow-up. The change in serum creatinine over time was also not significantly different within each group (SF, p = 0.09; IF, p = 0.38). CONCLUSIONS: In this study, suprarenal fixation was associated with a very small incidence of immediate renal and visceral artery occlusion. There did not appear to be any medium-term sequelae of suprarenal fixation. PMID- 17401762 TI - Pharmacological probes to measure the importance of the autonomic nervous system. PMID- 17401763 TI - A new model for American health care: transformation for a dysfunctional system. PMID- 17401764 TI - I Can PresCribE A Drug: mnemonic-based teaching of rational prescribing. PMID- 17401765 TI - Neonatal circumcision model and competency evaluation for family medicine residents. AB - Neonatal circumcision is one of the most common procedures performed by family physicians and is a recommended procedure for resident training. Few models or simulators exist. A model of neonatal genitalia was constructed using a cocktail wiener covered by a surgical glove finger. A competency checklist was used to review the steps of the procedure. Posttest/pretest-posttest assessment found significant improvement in knowledge and comfort in three groups of learners. Ninety percent of participants were deemed competent in all 15 areas of the checklist. The easily replicable model was felt to reasonably model the neonatal genitalia for circumcision training. PMID- 17401766 TI - Movies and emotional engagement: laughing matters in lecturing. PMID- 17401767 TI - A grand seduction indeed. PMID- 17401768 TI - Influence of hometown on family physicians' choice to practice in rural settings. AB - OBJECTIVE: This study examines the influence of a physician's hometown location on the choice of practice location, adjusting for confounding variables. METHODS: Medical school records for 2,487 Indiana University graduates (classes of 1988- 1997) were matched to the American Medical Association's Masterfile data to identify the graduates' current practice locations and specialties. Urban influence codes were assigned to each county in Indiana for the purposes of defining metro or nonmetro locations. Physician practice locations were mapped using ArcGIS software. Chi-square tests, logistic regression, and analysis of variance were used to examine the influence of hometown on choice of practice location. RESULTS: Chi-square tests revealed significant associations between physician hometown and current practice location. Logistic regression, controlling for age and gender, predicted physicians (all specialties) from nonmetro hometowns were 4.7 times as likely to locate their practice in a nonmetro location as compared to their peers from metro hometowns. Similarly, family physicians from nonmetro hometowns were 4.4 times as likely to choose a nonmetro practice location. There was not a significant difference in the mean distance between hometown and practice location for physicians from nonmetro hometowns compared to those from metro hometowns. CONCLUSIONS: This study underscores the influence of physicians' hometown on their choice of practice location. PMID- 17401769 TI - The effect of offering international health training opportunities on family medicine residency recruiting. AB - BACKGROUND AND OBJECTIVES: While medical students' interest in family medicine declines, and residency programs face recruiting challenges, interest in international health is increasing. We studied the influence of offering an international health track (IHT) on residency recruitment. METHODS: We surveyed all graduates between the years 1994--2003 of a family medicine residency program offering an optional IHT (n=90). Descriptive and bivariate analyses characterized and compared the geographic scope and residency selection criteria of IHT participants and nonparticipants. RESULTS: The response rate was 77%. Compared to nonparticipants in the IHT, residents who participated in the IHT reported coming a significantly greater median distance from their medical school training site (250--499 miles versus 0--99) and from their "home" (500--999 miles versus 100- 249) for residency training. Participants reported that the factor most positively influencing their choice of this residency training site over others was the IHT (mean=+2.5, standard deviation [SD]=0.90), outscoring, on average, location, faculty quality, resident quality, and spousal preference. Nonparticipants' choices were influenced most by the traditionally reported selection factors location (mean=+2.37, SD=1.14) and faculty quality (mean=+2.29, SD=0.97). CONCLUSIONS: One program's experience suggests that the presence of international health training opportunities may influence the residency selection choices of family medicine residents. As family medicine moves to reinvent residency education and to recapture student interest, attractive training models that integrate clinical, community, and public health should be evaluated further. PMID- 17401770 TI - The use of standardized patients to evaluate family medicine resident decision making. AB - BACKGROUND AND OBJECTIVES: This study was intended to assess the usefulness of standardized patients (SPs) in the evaluation of family medicine residents' clinical decision-making skills in ambulatory settings. METHODS: A pool of SPs was trained about the symptoms of one of three clinical conditions (depression, headache, or irritable bowel syndrome). These patients were then surreptitiously incorporated into the office hours of 11 residents on one occasion during their second year and once during their third year of training. A different SP was used at each encounter, but the same clinical case was presented each year for that particular resident. The SPs were given a questionnaire after each office visit to evaluate whether the resident completed elements of history, physical exam, clinical decision making specific to their case, and to report their satisfaction with the encounter. In addition, the assessment component of the resident's progress notes for each SP visit was reviewed by a single examiner to determine if the correct diagnosis was made by the resident. RESULTS: The residents showed a statistically significant decrease in the percentage of checklist items completed for all clinical cases from the second year (82.70%) to the third year (75.55%). However, the average patient satisfaction was unchanged, as was the number of correct diagnoses, even though fewer questions were asked. CONCLUSIONS: The use of SPs is a feasible and potentially useful method for evaluating family medicine resident decision making. Several factors may account for the differences in resident performance with SP scenarios. PMID- 17401771 TI - A comfortable relationship: a patient-derived dimension of ongoing care. AB - BACKGROUND AND OBJECTIVES: Continuity of care is considered essential to primary care and improves health outcomes. However, it is unclear whether patients and physicians define this concept similarly. This study examined how patients perceive a continuity relationship, from its development through its consequences. METHODS: This was a qualitative study using grounded theory methodology. Researchers selected and interviewed a purposeful sample of 14 primary care patients. Audiotapes of the interviews were transcribed and analyzed using open, axial, and selective coding schemes. RESULTS: Although the majority of patients were not familiar with the phrase continuity of care, all patients in the study identified comfort with their doctor as important to establishing and maintaining an ongoing relationship. Comfort with a physician was developed in several ways. Patients described their level of comfort with a physician as influencing making and keeping appointments, the likelihood that they would discuss concerns about sensitive issues, their perception that medical problems were resolved faster, and their trust in the physician's treatment plan. CONCLUSIONS: For patients, comfort appears to be an important dimension of the physician-patient continuity relationship. Primary care physicians and researchers should further explore how to use patient comfort as an element of care. PMID- 17401772 TI - The use of an electronic medical record to improve documentation and treatment of obesity. AB - BACKGROUND AND OBJECTIVES: Physicians underdocument and undertreat obesity. Electronic medical records (EMR) reminders have improved other preventive services such as cancer screening and immunizations. We explored whether an EMR automatic calculation of body mass index (BMI) improved clinician documentation and treatment of overweight and obesity. METHODS: We conducted a retrospective cross-sectional study of randomly selected patient charts before and after the implementation of an EMR. The primary outcome was documentation of overweight or obesity in the assessment/plan section or problem list section of the record. The secondary outcome was evidence in the medical record of treatment (defined as nutrition or exercise counseling or referral to a nutritionist) of overweight/obese patients. RESULTS: Documentation of obesity (BMI ? 30) improved from 31% to 71% (prevalence ratio [PR]=2.30, 95% confidence interval [CI]=1.44- 3.68) of obese patients after the implementation of an EMR calculation of BMI. Documentation of treatment of obese patients also improved, from 35% to 59%, (PR=1.84, 95% CI=1.19--2.86), but documentation and treatment of overweight patients (BMI>25<30) did not significantly improve. CONCLUSIONS: These findings support the effectiveness of an EMR automatic BMI calculation to improve documentation and treatment of obese patients but suggest that software modification and alternative strategies are needed to improve documentation and treatment for overweight patients. PMID- 17401773 TI - What should physicians know about hypertension? The implicit knowledge requirements in the maintenance of certification self-assessment module. AB - BACKGROUND AND OBJECTIVES: The American Board of Family Medicine (ABFM) Maintenance of Certification process requires family physicians to have a core knowledge base in key areas such as hypertension and diabetes care but does not define this knowledge in specific terms. We developed a method of content analysis to evaluate what type of knowledge is assessed on the ABFM's Hypertension Self-assessment Module (SAM) to better understand what the implied knowledge of a family physician should be. METHODS: In this qualitative descriptive analysis, we categorized the 60 questions comprising the knowledge assessment portion of the Hypertension SAM, version 2.20.03, into diagnosis, treatment, or etiology/general knowledge questions. Diagnosis and treatment questions were graded for relevance to typical family practice. Diagnosis questions were coded regarding importance. Treatment questions were subdivided into drug or nondrug treatments. Drug treatment items were categorized as testing knowledge of safety/tolerability issues, effectiveness issues, or cost considerations. RESULTS: The 60 questions represented 213 specific items of knowledge. Most (71%) of the items on the SAM focused on therapy, with the remainder evaluating knowledge of diagnosis issues or general knowledge. Of the therapy-related items, the items were evenly split between knowledge of safety/tolerability and knowledge of effectiveness (47.1% each). The remaining items required knowledge of nondrug therapy. No items evaluated knowledge of the relative cost of treatment or cost-effectiveness. With regard to the relevance of the tested information, only 70% of the items test knowledge that would be commonly needed in the practice of family medicine. CONCLUSIONS: There is currently no consensus on the discrete set of skills and knowledge that should be held by a competent family physician. In the absence of a comprehensive set of goals and objectives, the knowledge content being assessed in the SAMs can at least inform teaching programs about what their learners will be required to know to maintain certification. For the content area of hypertension, most of the knowledge required was regarding drug treatment. Interestingly, 30% of the knowledge content being assessed was found to be neither important nor commonly needed in the care of patients. We recommend that more work be done to define the specific knowledge and skills required for a competent family physician and that future maintenance of certification modules be written to assess mastery of these core requirements. PMID- 17401774 TI - New model innovation through faculty champions. PMID- 17401775 TI - International family medicine education. PMID- 17401778 TI - [Spanish adaptation and validation of the brief form of the Life Skills Profile 20: an instrument to assess daily living skills in real clinical settings]. AB - INTRODUCTION: The Life Skills Profile (LSP) is an assessment instrument of general function in activities of daily life, especially developed for people diagnosed of a severe mental illness. There are different versions of the LSP. The aim of this paper is to validate and adapt the brief form of the LSP (LSP-20) to the Spanish population. METHODS: Two different samples have been recruited for this study. The first sample was made up of 231 outpatients with a clinical diagnosis of schizophrenia who were randomly selected. The following analyses were performed: principal axis factoring analysis, internal consistency and convergent and discriminant validity between the LSP-20 and the Positive and Negative Symptoms Scale (PANSS). The second sample was recruited in order to assess interrater reliability and was composed of 30 long-stay inpatients with a diagnosis of chronic schizophrenia. RESULTS: Factor analysis produced four factors accounting for 41.22 % of the total variance. The following labels were suggested for the factors: compliance, social initiative, self-care and antisocial behavior. Internal consistency ranged from 0.69 (antisocial behavior) to 0.79 (social initiative). Correlations between the LSP-20 and the PANSS were moderate and clinically significant. Interrater reliability ranged from 0.58 (to have defined interests) to 0.89 (to wear clean clothes usually). CONCLUSIONS: The brief form of LSP can be useful in real clinical settings to help to personalize the aims of the rehabilitation interventions and to evaluate the health care model. PMID- 17401779 TI - [Differences in management of depression in Spain from psychiatric and primary care physician point of view]. AB - INTRODUCTION: Given that depression is the most prevalent psychiatric disorder in our society, the current situation in pharmacotherapy of depression in Spain has been studied from the point of view of psychiatrists and general practitioners (GP). METHOD: A total of 339 interviews were carried out with two groups of physician (238 primary care physicians and 101 psychiatrists) from different Spanish cities. Distribution, application and questionnaires collection were made throughout 2002. RESULTS: The diagnostic instruments most commonly used by more than 90 % of both medical groups to detect a depressive disorder in a consultation are the evaluation of symptoms and the interview with the patient. However the main diagnostic problem was "masking" of depression with other symptoms/disorders. Most GP (95 %) and psychiatrists (99 %) establish a pharmacological treatment in all their depressive patients. Both groups coincide in most community use of SSRI as drugs of first choice (93% from both samples). The pharmacological agents most used by the psychiatrist are the serotonin selective reputake inhibitors (SSRI) (98.3 %), followed by venlafaxine (84.4 %), anxiolytics (68.4 %), mirtazapine (58.9 %) and reboxetine (55.8 %). In the case of GP, the most commonly used pharmacological groups are SSRI (98.3 %) and anxiolytics (73.4 %). In primary care, the SSRI are considered the most effective antidepressant group. However, the tricyclic antidepressives (TCA) would be the most effective for the psychiatrists. CONCLUSIONS: For psychiatrists and GP, the quality of care of depression in Spain is rated positively. However, there is a group of deficiencies and some aspects that need to be clearly improved, such as the time of consultations, coordination between GP and psychiatrists, waiting lists and available resources for mental health units. PMID- 17401780 TI - [Adaptation and validation of the Family Accommodation Scale for obsessive compulsive symptoms in a sample of Spanish adolescents]. AB - INTRODUCTION: Family influence and involvement in the obsessive- compulsive symptoms of their relatives are widely recognized in clinical practice although there is a very little research investigating those variables. The Family Accommodation (FAS) for obsessive-compulsive symptoms is an interview developed to evaluate those aspects of family interactions. The aim of this study is to present the Spanish translation, adaptation and validation of the FAS in a sample of Spanish adolescents. METHOD: This is a 12 month follow-up study of 20 adolescents diagnosed of obsessive-compulsive disorder (OCD), and their families, who started treatment in a child and adolescent mental health outpatient unit. RESULTS: The reliability measurement of the scale obtains good values (Cronbach's alpha is 0.87, Guttman split-half is 0.81). Measurement of convergent validity has good correlation levels with other measures of OCD symptom severity, both at the onset of the treatment as well as at the 12 month follow-up. There are no significant differences in FAS scores between families whose mother and/or father exhibit OCD symptoms of their own and those without this condition. The results support the hypothesis that the evaluation interview of the Spanish adaptation of the family accommodation/involvement in OCD symptoms, as the original, is a reliable and valid measure of family participation in obsessive-compulsive symptoms of adolescent with OCD. PMID- 17401781 TI - [Safety and tolerability of olanzapine versus risperidone: a one-year randomized study in outpatients with schizophrenia with prominent negative symptoms]. AB - OBJECTIVE: To evaluate the safety and tolerability of long-term treatment with olanzapine versus risperidone in schizophrenic outpatients with prominent negative symptoms. METHODS: This was a multi-center, randomised, open-label, parallel, dose-flexible, 1 year study of outpatients with schizophrenia (DSM-IV criteria) with prominent negative symptoms (SANS Global score > or =10). Safety was evaluated by recording treatment-emergent adverse events, vital signs, body weight and, when available, laboratory parameters. Extrapyramidal symptoms (EPS) were evaluated by a questionnaire based on the UKU scale, and sexual dysfunction by the Psychotropic-Related Sexual Dysfunction Questionnaire (PRSexDQ). RESULTS: The mean (+/- SD) modal dose throughout the study was 12.3 (+/- 6.3) mg/day for olanzapine and 5.2 (+/- 2.5) mg/day for risperidone. EPS were significantly more frequent in the risperidone-treated patients 50.4 % versus 28.9 % for olanzapine (p = 0.0006). Olanzapine patients showed significantly greater reductions (improvement) from baseline in the PRSexDQ score (p=0.0292) and risperidone patients reported significantly more sexual adverse events (21.1% versus 7.3% for olanzapine; p=0.0018). Mean body weight gain was not significantly different at endpoint (3.5 kg gained with olanzapine versus 1.9 kg gained with risperidone; p=0.3522), but the proportion of patients showing a body weight increase > or =7% was higher among the olanzapine-treated patients (37.8% versus 16.8%; p=0.0012). CONCLUSIONS: Significantly less treatment-emergent extrapyramidal and sexual adverse events were observed in patients treated with olanzapine compared to those treated with risperidone. Mean body weight increases with both drugs were not significantly different after one year. Olanzapine patients presented a significantly higher incidence of clinically important body weight increase when compared with patients treated with risperidone. PMID- 17401782 TI - [Organic personality disorder: conceptual review and research strategies]. AB - The historical review of "psychiatric personality disorders" reveals the lack of convergence of those disorders with the organic personality disorder (OPD). Only the psychopathy concept has been used as a psychopathological phenotype for one of the groups of OPD, the so-called "pseudopsychopaths". These patients have been described from the beginning of the XXth century under the heading of "frontal lobe syndrome". It was only with the development of the psychiatric nosologies, towards the middle of the XXth century, that the term "organic personality disorder" started to be used. The accumulation of knowledge about the different prefrontal areas and the development of neuropsychological models that try to explain social behavior have opened new ways of understanding this syndrome. The orbitofrontal cortex has been identified as one of the key structures in behavioral and emotional regulation. Recognition of emotions in voices and faces, empathy, appreciation of humor, tasks that show "theory of mind" are some of the dimensions included in the examination of the non-cognitive functions of the prefrontal cortex PMID- 17401783 TI - [Endocannabinoid system and CNR1 gene polymorphisms in schizophrenia and addictive disorders]. AB - Substance abuse is the most prevalent comorbid psychiatric condition associated with schizophrenia. Cannabis is a drug frequently used for schizophrenic patients. In the last decades the endocannabinoid system and their endogenous ligands have been discovered. Endogenous cannabinoids act in the brain on cannabinoid CB1 receptor. On the other hand this system may be involved in several brain functions through neuromodulation dopaminergic and other neurotransmitter system involved in schizophrenic and substance abuse disorders. Advances of genetic research have addressed the focus on the search of candidate genes for both disorders. In this review we have summarized the studies published about the CNR1 gene on schizophrenia and substance abuse disorders. PMID- 17401784 TI - [Revision of the primary care version of the ICD-10. Mental disorders]. AB - Although the difficulty of applying psychiatric classifications to primary care has been widely criticized, there have been few investigations up to now to define and systematize the real demands in regards to these nosological systems. Recently, the revised version of the Mental and Behavior Disorders Chapter of the ICD 10 has been published. The new tool is the result of an elaboration process mainly developed by a group of 971 primary care physicians coordinated by 55 psychiatrists. The project was organized into three phases: a) evaluation of the current version and collection of proposals for change; b) definition of objectives for an optimized version; and c) writing a proposal of revised text. The result is a text that is more assimilable to a diagnostic and therapeutic guide than a mere coding system, more adapted to the role that the primary care physician can play in each disorder, more up-dated (especially in the treatment section) and more specific in many aspects. PMID- 17401785 TI - [Course and prognosis of a case of central pontine myelinolysis in eating behavior disorder]. AB - Central pontine myelinolysis (CPM) is a serious disorder that has been described in multiple diseases, generally involving important metabolic and hydroelectrolyte alterations. Although initially, its prognosis was usually fatal, there are a growing number of cases where the clinical symptoms begin abruptly and end after a short period, albeit with a persistence of the neuroimaging lesions. The case of a 22 year-old woman with a 6 year history of serious eating disorder with important physical deterioration and neurological and psychiatric symptoms suggestive of CPM is described. Despite the confirmation of the brain lesions through magnetic resonance imaging, neurological and psychiatric symptoms fully disappeared within a few weeks while the typical lesions of CPM remained. Although the risk of appearance of CPM exists during the course of an eating disorder, its prognosis does not seem to be as fatal as it was previously thought. Close monitoring of the clinical symptoms and neuroimaging findings should be carried out in these patients during the first months. PMID- 17401786 TI - [Olanzapine-associated neuroleptic malignant syndrome. A case report and favorable response to risperidone]. PMID- 17401787 TI - [Severe abstinence due to Zolpidem]. PMID- 17401788 TI - Psychotropic drugs for terminally ill patients with respiratory disease. AB - Utilization of psychotropic drugs including hypnotics, sedatives, and parenteral morphine for terminally ill patients with respiratory disease has not been reported precisely. To better understand these drugs for dying patients with respiratory disease, we focused on the last month of life of them. A chart review, which was focused on symptoms and managements, in the last month of life of 337 patients who died of respiratory disease between April 2000 and March 2005 were performed. Hypnotics were prescribed in 35.8% and 23.2% of patients with malignant and non-malignant disease, respectively. Sedatives such as haloperidol and midazolam were utilized in 34.4% of patients with malignant disease, and 30.4% of those with non-malignant disease. Sixty-seven percent of patients with malignant and 22.4% of those with non-malignant disease had parenteral morphine. In patients with malignant disease, three of the main reasons for administration of morphine were pain, dyspnea, or terminal restlessness. In all of the patients with non-malignant disease, however, parenteral morphine was prescribed for the treatment of dyspnea. Our results showed that psychotropic drugs and parenteral morphine are required in some of terminally ill patients with respiratory disease. Although careful individualization of medication is appropriate, guideline for the prescribing psychotropic drugs for these patients will be required. PMID- 17401789 TI - [The factors affecting Beck depression scale in asthmatic patients]. AB - Symptoms of asthma can be intensified and/or mixed with depression since depression can cause asthmatic symptoms like complaints even in normal people. In the present study, depression index and its influencing parameters were examined in asthmatic patients. A questionnaire including sociodemografic features was applied to 120 patients, accepted as asthmatic according to the Thoracic Society Guidelines, who were following up in our asthma and allergy outpatient clinic. There were 98 female and 22 male patients and the mean age was 38.19 +/- 10.99. Physical examination, PEF and spirometric measures were made in all patients. Also daytime and nighttime symptom score, Q score, Beck depression score were applied to patients. Beck Depression Index (BDI) was in range of 0 and 37 and the mean was 11.26 +/- 8.54 (mild). According to the cut-off values, BDI scores were obtained as absent, mild, moderate and severe, 59%, 27%, 9%, 7% respectively. The mean BDI score was higher in female patients (12.32 +/- 8.77) than in male patients (6.55 +/- 5.41) and in married patients (11.91 +/- 8.77) than in unmarried ones (7.56 +/- 6.00). According to the results of FEV(1) values, PEF changes, there were no significant difference obtained between groups (p> 0.05). The mean value of BDI was found to be significantly high in patients with moderate and high daytime and nighttime score and Q score (p< 0.05). In conclusion, we think that, while evaluating the severity of asthma, if the spirometric results are not correlated with the symptoms then psychological conditions of the patients should be taken into consideration. PMID- 17401790 TI - [Factors affecting incidence of tuberculosis in Diyarbakir]. AB - Last decade, there have been important improvements about tuberculosis (Tbc) in the world. Today, 32% of world populations are infected by Tbc basil's, every year about 9 million people have been catching Tbc. We have purposed to determine the incidence of Tbc and the factors affect it, using rate of bacteriology in diagnosis, rate of treatment completion and to constitute an idea for innovation. Diyarbakir, standard monthly data forms used for informing of Tbc in tuberculosis control dispensary are investigated retrospectively among 1996-2004. The mean incidence of yearly Tbc is 37.77/100.000 and pulmonary Tbc is 30.11/100.000. In this period have been determined 3724 new Tbc patient; 2969 (79.7%) are pulmonary Tbc and 755 (20.3%) extrapulmonary Tbc. 842 patient (22.6%) of pulmonary Tbc are smear positive. Tbc pleurisy is the most common type of extrapulmonary Tbc in our series. 3354 (90.1%) of whole patients and 2624 (88.4%) with pulmonary Tbc had completed treatment. There is no data about the rate of cure. This is the most common problem Tbc control system. In 2005, cure rates will have been determined by the way of new form. 162 (4.4%) patient had abandoned treatment. Incidence of Tbc is higher than the incidence of Turkey. It was considered that at the first place highly growing population, the crowded family pattern sharing the same house and bad socioeconomic factors have been playing a major role. Furthermore, it was found that the rate of bacteriological diagnosis was low. In order to increasing of this rates should be carried out necessary studies, should be tried to diagnose all patients with bacteriological methods. End of the treatment cure should be tried to demonstrate by way of examination of sputum. The treatment process should be pursued by directly observed treatment strategy. PMID- 17401791 TI - [Diagnosis of venous thromboembolism by single-detector row and multi-detector row CT angiography]. AB - Aim of this study is to determine the diagnostic value of pulmonary angiography performed by either single-detector row CT (SDCT) or multi-detector row CT (MDCT) in patients suspected of venous thromboembolism (VTE). SDCT was performed on 36 and MDCT on 18 of total of 54 patients on whom V/Q scintigraphy was obtained with a suspicion of VTE. Sixteen out of 54 cases got additional pulmonary DSA. Statistical analyses were based on final clinical diagnoses of the individual cases. Twenty-six out of 54 cases in the study got the final diagnoses of VTE and VTE was certainly excluded in the remaining 28. Sixteen out of true 26 VTE cases were in the SDCT group while the other 10 cases took place in the MDCT group, none of which was missed by either technique. There was one false positive result in the SDCT group and none in the MDCT group (96% and 100% specificity respectively, 100% sensitivity for both). Only 9% of all pulmonary emboli detected by SDCT assisted pulmonary angiography were located in subsegmental arterial branches, whereas 24% of emboli detected by MDCT angiography were subsegmental. Both SDCT and MDCT angiography are reliable tests in the detection of VTE. MDCT assisted pulmonary angiography is superior than SDCT assisted pulmonary angiography in subsegmental VTE detection. PMID- 17401792 TI - [The effectiveness of thrombolytic therapy in patients with massive and submassive pulmonary thromboembolism]. AB - Thrombolytic therapy is the most effective therapy for massive pulmonary embolism (PTE). In this study we evaluated the symptoms, clinical and radiologic features and response to thrombolytic therapy in patients who had massive or submassive PTE. Thrombolytic therapy was administered for a mean period of 3 (1-5) hours to 19 patients with a mean age of 63.7 years who had the diagnosis of PTE based on symptoms which lasted for an average of 72 (2-240) hours and findings of echocardiography, spiral computed tomography (CT) angiography and perfusion scan. The patients to whom heparin infusion and oral anticoagulant treatment were given after thrombolytic therapy were evaluated. Bleeding as a complication was noted in six patients after thrombolytic therapy. Two patients died due to this complication. Control spiral CT angiography was performed to 12 of 15 (80%) patients who were initially diagnosed PTE by spiral CT angiography. While marked regression was noted in 5 (41.7%) patients in the early phase (second week), in 7 (58.3%) patients in the late phase (sixth month) PTE findings were completely disappeared. The median value of pulmonary artery pressure was 65 (45-70) mmHg before and 39.5 (30-45) mmHg after the treatment. Of the genetic factors studied before thrombolytic therapy, antithrombin III deficiency was found as the most common one. This study demonstrates that spiral thorax CT angiography is a very accurate diagnostic tool for the definitive diagnosis and transthoracic echocardiography is very useful for the rapid diagnosis and to decide for thrombolytic therapy, in especially patients who are not hypotensive and have submassive to massive PTE, and support the idea that thrombolytic therapy is life saving after reaching the certain diagnosis. PMID- 17401793 TI - [The smoking habits of health care workers and their approach to smoking in Gaziosmanpasa and Eyup counties of Istanbul]. AB - We aimed to determine the smoking habits of the health care workers and their approach to smoking. A questionnaire was applied to the health care workers about smoking in the counties of Istanbul (in Eyup and Gaziosmanpasa). Of 629 participants, the ratio of smoking in all health care workers was 49.3%. There was no difference in the frequency of smoking in the health care workers about sex, marital status, private or state hospital, the group of job, general practitioner or specialist doctor (p> 0.05). However, the rate of smoking was lower in married doctors than unmarried ones, and it was higher in male technician than females (p< 0.05). Partners of smoker doctors and nurses had higher smoking rate (p< 0.01). There was no difference between smoker and nonsmoker doctors in asking about smoking to their patients. The rate of "Yes" answer to the question of "Do you explain the necessity and recommend to give up smoking to more than 70% of your smoking patients?" asked to the doctors was different between smoker and nonsmoker doctors (58.8% and 72.5% respectively, p< 0.05). These results show that more efforts are needed and should be spent on the education about the harmful effects of tobacco products and needs of fighting against smoking in medical faculties, in health occupation schools, in all health care services. PMID- 17401794 TI - The cost of lung cancer in Turkey. AB - The aim of this study was to evaluate the individual and societal burden of lung cancer in Turkey. A total of 103 cases with lung cancer attended our department between January 2002 and February 2003 were included in our study prospectively. The primary outcome measure was the cost of disease until death of the patients or the end of study. All the costs were expressed as United States dollars (USD) and were estimated regarding the effective exchange rate at the time of recording. Descriptive statistics, chi-square, Fisher's exact test, Kaplan-Meier analysis and non-parametric "Bootsraping" tests were performed to evaluate the data. The average survival was 6.8 months. The estimated total direct cost for the entire group was 564.490 USD, and the direct cost per patient was 5.480 +/- 4.088 USD. The total cost of lung cancer in the study group was 1.473.530 USD, with a per-patient cost of 14.306 +/- 17.705 USD. The average direct cost per life year was 18.058 +/- 25.775 USD. Age, gender and histopathology did not affect the cost, whereas direct medical costs were increased with increasing stage. With the low life expectancy and cure rates, lung cancer has been alerting for the cost minimization and disease control measures. PMID- 17401795 TI - [Does tumor type and sex distribution of primary lung cancer change? The comparison of the results of 2004 and previous years]. AB - The aim of the present study was to evaluate the tumor cell type and sex distribution of patients diagnosed with primary lung cancer during 2004. Patients with primary lung cancer were detected from pathology records. Clinical files of patients were analyzed retrospectively. 1403 patients with primary lung cancer were included in the study. 1238 (88.2%) patients were male and 165 (11.8%) were female and female to male ratio was 1/7.5. When the ratio was compared with 1/10.9 in 1998, the ratio of female patients was determined to increase. Tumor cell type was squamous cell carcinoma in 577 (41.1%) patients, adenocarcinoma in 359 (25.6%), small cell carcinoma in 184 (13.1%), nonsmall cell carcinoma in 115 (8.2%) and other malign tumors in 21 (1.5%) patients. Tumor cell type was not detected in 147 (10.5%) patients. When 147 patients were excluded from the study, the frequency of squamous cell carcinoma was 45.9%, adenocarcinoma was 28.6% and small cell carcinoma was 14.6%. The results of our study show that squamous cell carcinoma is the most frequent tumor cell type in our center and the ratio of female patients is increasing. PMID- 17401796 TI - [The effect of spinal anesthesia on pulmonary function tests in old patients]. AB - Pulmonary function test (PFT) results are mainly dependent on age, sex, height, weight, pulmonary mechanics disturbances and cooperation of the subjects. The position and anesthesia type may also influence the PFT results. In this study we aimed to evaluate spirometric changes in old and young patients who performed spinal anesthesia. Fifty patients performed spinal anesthesia were randomized in two groups: Group 1 (n= 25) aged 60-85 years old and group 2 (n= 25) aged 20-59 years old. After electrocardiography, noninvasive blood pressure and peripheral oxygen saturation (SpO2) monitorization, spinal anesthesia using 0.5% hyperbaric bupivacain from L 3-4 intervertebral space was applied. Sensory block levels, hemodynamics and PFT such as forced vital capacity (FVC), forced expiratory volume/1 second (FEV(1)), peak expiratory flow (PEF), and forced expiratory flow at the 25 and 75% of the pulmonary volume (FEF(25-75)) were performed before and after spinal anesthesia in 10th, 40th and 100th minutes in supine and 30 degrees head position using hand type spirometry. Wilcoxon paired two tests statistical analysis was used to compare PFT changes of the subjects. Mean arterial blood pressure levels and spirometric measurements of FVC, FEV(1) and FEF25-75 decrease with respect to basal values in 40th minutes was significant in old patients whom spinal anesthesia was over Th6 level but in young patients the changes were not significant. PFT decrement probabilities should be taken in account in old patients supposing for spinal anesthesia and be paid attention for high level spinal blocks in risk group patients. PMID- 17401797 TI - [The role of video-assisted thoracoscopic surgery in the treatment of pleural empyema]. AB - Pleural empyema is a serious problem which affects any age group and still there is no standard approach. Our purpose in this study was to assess the safety, feasibility and efficacy of the video-assisted thoracoscopic surgery (VATS). Between 1997 and 2004 83 patients with empyema were enrolled in this study. Fifty four patients were male, 29 females and the mean age was 39.4. The mean duration time of symptoms before VATS was 27.3 days. Twenty-two patients were in group-5, 44 in group-6 and 17 in group-7 according to Light's classification of parapneumonic effusions and empyema. Twenty-one patients underwent chest tube before VATS because of their conditions. VATS was converted to thoracotomy in 13 patients. All patients underwent debridement and irrigation during VATS. The cure was achieved in 58 (69.9%) patients with the mean 7.63 days drainage time. Twelve patients were underwent open decortication in the follow-up period. Ten of them were group-7 and 2 in group-6 (p= 0.006). The diagnosis was achieved in 60 (72.3%) patients as nonspecific pleuritis, tuberculosis in 19 and various diagnosis in 4. There was no mortality and minor complications occured in 14 patients. VATS is a safety and efficacy procedure in the treatment of early stage empyema. Use of VATS in early period of multiloculary empyema, prevents patients from unnecessary thoracotomies. PMID- 17401798 TI - Idiopathic pulmonary hemosiderosis in an adult patient responded well to corticosteroid therapy. AB - Idiopathic pulmonary hemosiderosis (IPH) is a very rare disorder of unknown etiology characterized by recurrent or chronic hemorrhage and accumulation of hemosiderin in the lung parenchyma. It is most common in children but can occur in adults. Clinical manifestations of the disease include iron deficiency anemia without any known cause, pulmonary symptoms such as hemoptysis, dyspnea and cough, and parenchymal lesions on chest X-ray. The clinical course of the disease may vary from patient to patient however, in general, the prognosis of the disease is worse. Treatment is symptomatic and supportive. Corticosteroids and other immune suppressive agents were used for the therapy of IPH. Since it is seen rarely in adults and the clinical course of the disease vary from patient to patient we presented an adult male patient with IPH responded well to steroid therapy clinically and radiologically. PMID- 17401799 TI - A new non-invasive diagnostic method for lung lobe torsion: pulmonary CT angiography. AB - Torsion of the remaining lung lobe after lobectomy is an uncommon event which is difficult to diagnose and may result in hemorrhagic infarction and fatal gangrene. We report a case of right upper lobe torsion after middle lobe resection for bronchiectasis, diagnosed with noninvasive pulmonary computerized tomography (CT) angiography. Rethoracotomy with an upper lobectomy was performed. She was asymptomatic at the two-year follow-up. Although the bronchoscopy has a great value in the diagnosis, CT angiography is an effective noninvasive method for confirming the correct diagnosis. A high index of clinical suspicion, early diagnosis and aggressive management may improve survival. PMID- 17401800 TI - Denim sandblasting and silicosis two new subsequent cases in Turkey. AB - In our country quartz is widely used for denim sandblasting and new cases with silicosis are defined. Silicosis is a preventable occupational lung disease caused by inhaled dust containing crystalline silica and no effective treatment for silicosis is available (1). CASE 1: A 23-years old man was admitted to the hospital with dyspnea on effort and cough. He had worked in a denim manufacturing factory for three years at sandblasting and dyeing. Physical examination revealed decreased breath sounds. Chest X-ray showed bilateral reticulonodular densities predominantly at middle and lower zones and minimal pneumothorax. Hypoxemia was determined in arterial blood gas analysis (BGA). Pulmonary function tests (PFTs) showed restrictive disorder. Lymphocytic alveolitis was demonstrated in the bronchoalveolar lavage fluid. Open lung biopsy revealed interstitial fibrosis and foreign particles which were seen more clearly under polarized light in interstitial areas. The patient was diagnosed as accelerated silicosis. CASE 2: A 25-years old man was admitted to the hospital with shortness of breath and haemopthysis. Two years ago, he had worked at the same place for 1.5 years. Bilateral reticulonodular densities at middle and lower zones and minimal unilateral pneumothorax were seen in chest X-ray. Restrictive disorder at PFT and hypoxemia in BGA were observed. Because of the similarity of complaints, radiological findings and occupational history with the former patient, no other further and invasive procedure was planned and the patient was diagnosed as accelerated silicosis. CONCLUSION: The usage of sandblasted denims increase recently and denim sandblasting is being frequently made especially in small work places. Since there is no definite treatment for silicosis, it is important to take necessary precautions to improve the conditions of the factories. PMID- 17401801 TI - Swallowing of a dynamic tracheal stent: an unusual complication. AB - Cicatricial tracheal stenoses are mainly related to iatrogenic injury as postintubation, posttracheotomy or postoperative. Advances in airway prosthetics have provided a variety of silicone stents and expandable metal stents as a therapeutic option for tracheal stenoses. Nowadays, dynamic tracheal stents are performed prevalently for the treatment of tracheal stenosis. Four major problems are currently associated with airway stents: Mucostasis, formation of granulation tissue, infection and migration. In this report, we present a case of swallowing a tracheal dynamic stent as an unusual complication of tracheal stents. PMID- 17401802 TI - A rare presentation of hydatid cyst in a child. AB - Cyst hydatid is the most widespread, serious cestode infection in the world. The most common organ affected by hydatid disease is the liver followed by the lungs and the two organs are affected simultaneously in about 5-13% of cases. The involvement of kidneys are rarely reported. Here we reported a patient with hydatid cysts in lung and kidney which the surgery was effective for both organs. The rarity of this case is the unusual combination of the cyst development in these organs without the involvement of liver. PMID- 17401803 TI - A squamous cell lung carcinoma with abscess-like distant metastasis. AB - This is a metastatic spread of squamous cell lung carcinoma to lungs, liver, lymph node, bone and subcutanous region as multiple abscess-like lesions. A fifty five years old man admitted to the out-patient clinic with fever, cough, hemopthysis, night sweats, chest pain, abdominal pain and weight loss. In a short period of time abcess like lesions developed in his lungs, liver, lymph node, bone and subcutanous region. Though the clinical presentation is suggestive for an infectious condition, no success to antimicrobial treatment and negative results of microbiological studies have arised a need to further investigations. Histopathological studies of the abscess wall ultimately gave the definitive diagnosis as metastatic squamous cell carcinoma. We believe that case report is interesting because of the uncommon metastatic lesions masquerading the abscesses and also wide-spread multiple distant invasions of a squamous cell lung carcinoma in a short time period. PMID- 17401804 TI - [Bird fancier's lung disease (report of two cases)]. AB - Bird fancier's lung disease (BFLD) is a hypersensitivity pneumonia which develops in response to organic bird products. Two patients (25 y/M, 43 y/F) were admitted to our clinic with complaints of dyspnea, fewer and weight loss, both had history of pigeon exposure and we investigated them for BFLD. First patient had restrictive pulmonary function tests with DLCO of 49%, and the second patient had obstructive pulmonary function tests with DLCO of 33%. HRCT of both patients revealed nodular infiltrations. With these findings and history of pigeon exposure (the first patient at his job, the second patient at home) the diagnosis of BFLD was established. First patient had complete remission after removal of pigeons from the environment. Initial inhaled corticosteroid and later systemic corticosteroid therapy was given to second patient. In conclusion, if BFLD, a rare disease, is considered initially in the differential diagnosis and environmental exposure is evaluated, the diagnosis and treatment is possible. PMID- 17401805 TI - [New diagnostic tool of tuberculosis: interferon-gamma assays]. AB - New developments have been performed in diagnosis of the tuberculosis (Tbc) which is still the most common and the most mortal preventable infectious disease. Tuberculosis skin test (TST) an immunologic diagnostic method has been used in the diagnosis of Tbc ever since, and is still considered as a gold standart for diagnosis of latent Tbc infection but so far has lost its credibility as a diagnostic test due to its false positivity and negativity characteristics and due to this reason a more sophisticated diagnostic method search is underway. An alternative to the TST has emerged in the form of a new type of in-vitro T-cell based assay: The interferon-gamma (IFN-?) assay. IFN-? assays are based on the principle that T-cells of individuals sensitised with Tbc antigens produce IFN-? when they reencounter mycobacterial antigens. In this article the principles leading to the production of IFN-? has been reviewed and its usage in real practise has been discussed. PMID- 17401806 TI - 'Silicosis' over again; causes and responsibilities. PMID- 17401807 TI - Genetic structure of Araucaria angustifolia (Araucariaceae) populations in Brazil: implications for the in situ conservation of genetic resources. AB - The distribution of the genetic variation within and among natural populations of A. ANGUSTIFOLIA growing in different regions in Brazil was assessed at microsatellite and AFLP markers. Both markers revealed high gene diversity ( H = 0.65; AR = 9.1 for microsatellites and H = 0.27; P = 77.8 % for AFLPs), moderate overall differentiation ( RST = 0.13 for microsatellites and FST = 0.10 for AFLPs), but high divergence of the northernmost, geographically isolated population. In a Bayesian analysis, microsatellite data suggested population structure at two levels: at K = 2 and at K = 3 in agreement to the geographical distribution of populations. This result was confirmed by the UPGMA dendrogram based on microsatellite data (bootstrap support > 95 %). Non-hierarchical AMOVA revealed high variation among populations from different A POSTERIORI defined geographical groups. The genetic distance between sample locations increased with geographical distance for microsatellites ( R = 0.62; P = 0.003) and AFLPs ( R = 0.32; P = 0.09). This pattern of population differentiation may be correlated with population history such as geographical isolation and postglacial colonization of highlands. Implications of the population genetic structure for the conservation of genetic resources are discussed. PMID- 17401808 TI - Equivalence of three allocation currencies as estimates of reproductive allocation and somatic cost of reproduction in Pinguicula vulgaris. AB - Which is the most appropriate currency (biomass, energy, water, or some mineral nutrient) for expressing resource allocation in plants has been repeatedly discussed. Researchers need to assess to which extent interindividual, interpopulational, or interspecific comparisons of resource allocation could be affected by the allocation currency chosen. The "currency issue" is relevant to at least three related aspects of resource allocation to reproduction: (a) reproductive allocation (RA), (b) size-dependence of reproductive allocation, and (c) somatic cost of reproduction (SCR). Empirical tests have mostly dealt with the first aspect only. We examined the equivalence of estimates for the three aspects above across three different allocation currencies (dry mass, N, P) in 11 populations of PINGUICULA VULGARIS. For RA we studied the equivalence of allocation currencies at three scales: among individuals of the same population, between populations of the same species, and among species. Equivalence of currencies in the ranking of RA for individuals within populations was high ( RS >/= 0.43) and did not strongly decrease when comparing populations or species. Excepting for size-dependence of RA, ranking of RA, or SCR between populations was equivalent for biomass and N, but not for P. Our study gives two positive guidelines for empirical plant reproductive ecologists facing the "currency issue": (1) become increasingly concerned about the "currency issue" as you increase the scale of your comparison from individuals to populations to species, and (2) avoid estimating allocation in redundant currencies (biomass and N in our case) and choose preferentially "complementary" currencies that provide a broader view of allocation patterns (biomass and P in our case). PMID- 17401809 TI - The impact of ozone on juvenile maize (Zea mays L.) plant photosynthesis: effects on vegetative biomass, pigmentation, and carboxylases (PEPc and Rubisco). AB - The impact of ozone on crops was more studied in C (3) than in C (4) species. In C (3) plants, ozone is known to induce a photosynthesis impairment that can result in significant depressions in biomass and crop yields. To investigate the impact of O (3) on C (4) plant species, maize seedlings ( ZEA MAYS L. cv. Chambord) were exposed to 5 atmospheres in open-top chambers: non-filtered air (NF, 48 nL L (-1) O (3)) and NF supplied with 20 (+ 20), 40 (+ 40), 60 (+ 60), and 80 (+ 80) nL L (-1) ozone. An unchambered plot was also available. Leaf area, vegetative biomass, and leaf dry mass per unit leaf area (LMA) were evaluated 33 days after seedling emergence in OTCs. At the same time, photosynthetic pigments as well as carboxylase (PEPc and Rubisco) activities and amounts were also examined in the 5th leaf. Ozone enhanced visible symptoms characterizing foliar senescence. Across NF, + 20, + 40, and + 60 atmospheres, both chlorophylls and carotenoids were found to be linearly decreased against increasing AOT40 ( CA. - 50 % in + 60). No supplementary decrease was observed between + 60 and + 80. Total above-ground biomass was reduced by 26 % in + 80 atmosphere; leaf dry matter being more depressed by ozone than leaf area. In some cases, LMA index was consistent to reflect low negative effects caused by a moderate increase in ozone concentration. PEPc and Rubisco were less sensitive to ozone than pigments: only the two highest external ozone doses reduced their activities by about 20 - 30 %. These changes might be connected to losses in PEPc and Rubisco proteins that were decreased by about one-third. The underlying mechanisms for these results were discussed with special reference to C (3) species. To conclude, we showed that both light and dark reactions of C (4) photosynthesis can be impaired by realistic ozone doses. PMID- 17401810 TI - Pollination biology and the impact of floral display, pollen donors, and distyly on seed production in Arcytophyllum lavarum (Rubiaceae). AB - In animal-pollinated plants, two factors affecting pollen flow and seed production are changes in floral display and the availability of compatible mates. Changes in floral display may affect the number of pollinator visits and the availability of compatible mates will affect the probability of legitimate pollination and seed production. Distyly is a floral polymorphism where long styled (pin) and short-styled (thrum) floral morphs occur among different individuals. Distylous plants frequently exhibit self and intra-morph incompatibility. Therefore changes in morph abundance directly affect the arrival of compatible pollen to the stigmas. Floral morph by itself may also affect female reproductive success because floral morphs may display differences in seed production. We explored the effects of floral display, availability of neighboring compatible mates, and floral morph on seed production in the distylous herb ARCYTOPHYLLUM LAVARUM. We found that floral display does not affect the mean number of seeds produced per flower. There is also no effect of the proportion of neighboring legitimate pollen donors on seed production in pin or thrum flowers. However, floral morphs differed in their female reproductive success and the thrum morph produced more seeds. Hand pollination experiments suggest that differences in seed production between morphs are the result of pollen limitation. Future research will elucidate if the higher seed production in thrum flowers is a consequence of higher availability of pollen donors in the population, or higher efficiency of the pin morph as pollen donor. PMID- 17401811 TI - Formation and distribution of sennosides in Cassia angustifolia, as determined by a sensitive and specific radioimmunoassay. AB - A radioimmunoassay for the quantitation of nanogram-amounts of sennoside B and related compounds in plant extracts is described. The assay makes use of [ (3)H] 8-glucosidorheinanthrone of high specific activity (5.2 Ci/mmol) whose synthesis is reported here. From this material, [ (3)H]-sennoside A and [ (3)H]-sennoside B have also been synthesized. The assay is applied to the analysis of sennoside formation and distribution in CASSIA ANGUSTIFOLIA VAHL. High levels of sennosides in dried leaves and fruits have been observed whereas the seed alone, as well as stems and roots, contain very little sennoside. In flowers, as much as 4-5% of the dry weight consists of sennoside B and other immunoreactive constituents. Sennosides have been found in cotyledons of three day old seedlings in concentrations comparable to that of the mature leaf. Upon dehydration, leaf levels of sennoside B rise steadily, this rise being inversely correlated with the water loss. The absolute levels of sennoside B formed this way are the same as compared to rapid drying at 60 degrees C. PMID- 17401812 TI - [On the active agents of valerian]. AB - C13-NMR-shifts of valepotriates show a quite characteristic dependence on type and position of the acyloxy substituents. The structure of well known and new valepotriates have been clarified. PMID- 17401814 TI - [Anaxagoreine, a new aporphine alkaloide, isolated from two species of the genus Anaxagorea]. AB - From several parts of two species of Anaxagorea two alkaloids asimilobine and anaxagoreine have been isolated. The last one, a new hydroxy-7 aporphine is characterised by an unusual configuration in the Annonaceae. PMID- 17401815 TI - Alkaloids of Colchicum visiani. AB - From the bulbs, flowers and flowerstalks of Colchicum visiani Parl. of Yugoslavian origin, were isolated and identified the neutral-phenolic and alkaline alkaloids listed in Table II. This investigation showed that the alkaloid content of Colchicum visiani Parl. is similar to that of Colchicum autumnale L., with the exception of a tropolonic alkaloid hR (f)-16 (found as well in Colchicum dorfleri) and a tropolonic alkaloid hR (f)-13, which had so far been unidentified in Colchicum species. PMID- 17401816 TI - Meconic acid and alkaloids in Papaver somniferum and P. bracteatum. AB - Meconic acid (C (7)H (4) O (7)) has an unusually low H content in relation to C and O; it seems also to be restricted to Papaver species which characteristically produce morphinanes. The suggestion that it is an energy exhausted product of the citric acid cycle whose formation controls alkaloid production has been investigated. Using HPLC methods the concentrations of meconic acid and of alkaloids in the latex have been shown to vary significantly during the day suggesting active metabolism but little firm evidence of formation of meconic acid from fumarate or pyruvate was obtained. PMID- 17401817 TI - Alkaloids from Egyptian Papaver rhoeas. AB - N-methylasimilobine was isolated as the major alkaloid from an Egyptian sample of P. RHOEAS L. Other alkaloids isolated from the same plant material were stylopine, coptisine, isocorydine and rhoeadine. Chromatography indicated the presence of at least nine other minor alkaloids. PMID- 17401818 TI - Studies on Indonesian medicinal plants VI further alkaloids from Fibraurea chloroleuca. AB - From the stem and root bark of Fibraurea chloroleuca Miers the alkaloids magnoflorine, pseudocolumbamine, dehydrocorydalmine and palmatrubine were isolated and identified by their spectral data. By means of TLC the alkaloids berberine and berberrubine were found to be present in minute amounts. PMID- 17401819 TI - The Alkaloids of Rauwolfia oreogiton. AB - 11 indole alkaloids were isolated from Rauwolfia oreogiton roots and identified. The alkaloids comprised dihydroindole, heteroyohimbine, oxindole, yohimbine and 18-hydroxy-yohimbine ester types. Chromatographic comparison of alkaloidal extracts from Rauwolfia oreogiton and Rauwolfia volkensii roots revealed no basis for chemical differentiation of the species. PMID- 17401820 TI - [Determination of structures by H NMR at 400 MHz: albifloranine, a new alkaloid from Tabernaemontana albiflora]. AB - The structure of albifloranine 1, a new alkaloid of ibogane type from the stem bark of Tabernaemontana albiflora (M IQ.) Pull. (Apocynaceae) has been determined. A detailed (1)H NMR study of albifloranine 1, coronaridine 2 and heyneanine 3 is presented. PMID- 17401821 TI - Iridoids, XXIX - Sinuatol (6-O-alpha-L-rhamnopyranosylaucubin) from Verbascum sinuatum. AB - From the aerial part of Verbascum sinuatum L. a new iridoid diglycoside III has been isolated and its structure elucidated as 6-0-alphaL-rhamnopyranosylaucubin. PMID- 17401822 TI - Iridoids from Premna latifolia. AB - Two iridoids, 7-deoxyloganic acid also known as bisdesoxydihydromonotropein and geniposidic acid, from Premna latifolia Roxb. (Verbenaceae) have been reported. PMID- 17401823 TI - [The cyanogenic glycosides of triticum, secale and sorghum]. AB - The cyanogenic glucosides obtained from seedlings of Triticum aestivum ssp. spelta, the inflorescences of Secale montanum and the caryopses of SORGHUM BICOLOR were identified by chromatographic and spectroscopic techniques. In all cases the cyanogenic principle is 2-beta- D-glucopyranosyloxy- 2 - (4 hydroxyphenyl) -2S-acetonitril (Dhurrin). According to thinlayer chromatographic investigations the cyanogenic glucoside of Secale cereale seems also to be Dhurrin. PMID- 17401824 TI - Effects of age and growth regulators on serially propagated Digitalis Ianata leaf and root cultures. AB - The cardenolide thin-layer chromatographic (TLC) and gasliquid chromatographic (GLC) patterns from leaf and root culture extracts were similar to those obtained from the plant. Leaf cultures at 8 and 12 wk-old contained more primary cardiac glycosides (Lanatoside A, B, C), than 4 wk-old leaf cultures. As high as 44 mg% dry wt. of digoxin, as determined by RIA, was present in 12 wk-old leaf cultures. The concentration of digoxin increased in both leaf and root cultures when treated with either gibberellic acid (GA) or mefluidide, and leaf organ cultures elongated with GA treatment. PMID- 17401825 TI - [Flavonoids in the flowers of Primula officinalis]. AB - From methanolic extracts of fresh flowers of Primula officinalis 19 flavonoids were isolated or detected: quercetin, luteolin, kaempferol, isorhamnetin, apigenin; quercetin 3-O-glucoside, -rutinoside, -robinobioside, -gentiobioside, (glucosyl-(1-->2) -glucosyl-(1-->6))-glucoside, -(rhamnosyl)-robinobioside; isorhamnetin 3-O-glucoside, -rutinoside, -robinobioside, -(rhamnosyl) robinobioside; kaempferol 3-O-rutinoside, -robinobioside; limocitrin 3-O glucoside; 3', 4'-dihydroxyflavonglucoside. Other compounds isolated were epicatechin, epigallocatechin, and proanthocyanidin B 2. PMID- 17401826 TI - Additional flavonoids from the leaves of Larrea tridentata. PMID- 17401827 TI - Investigation of erythrina spp. PMID- 17401828 TI - Alkaloids from Papaver armeniacum, P. fugax and P. tauricola. AB - Species from the section Miltantha are reviewed in the context of their contained alkaloids. Thebaine was isolated as the major alkaloid from a sample of P. fugax together with narcotine, pronuciferine, alpinigenine, O-ethylalpinigenine, amurensinine, N-methyl-crotonosine, armepavine, isocorydine and salutaridine as minor alkaloids. A second sample of P. fugax contained glaucamine and glaudine as major alkaloids with rhoeadine, oreogenine, oreodine and O-ethyl-glaucamine as minor alkaloids. Pronuciferine and armepavine were isolated as the major alkaloids from a sample of P. tauricola with narcotine, roemerine, nuciferine, nantenine and protopine as minor alkaloids. Another sample of P. tauricola yielded pronuciferine and mecambrine as major alkaloids with armepavine, lirinidine, thebaine and cryptopine as minor alkaloids. A sample of P. armeniacum contained rhoeadine and rhoeagenine as major alkaloids with lirinidine, cryptopine, glaudine and O-ethylrhoeagenine as minor alkaloids. Some 25 alkaloids representing 9 different alkaloidal-types were obtained from extracts of the three Miltantha species. The results show that at least three different chemical races of P. fugax and P. armeniacum exist in which either 1 benzyltetrahydroisoquinoline-, proaporphine-, aporphine- or morphinane- or rhoeadine- types are the major alkaloids. There are at least two different chemical strains of P. tauricola which contain either 1 beuzyltetrahydroisoquinoline-, proaporphine-, aporphine- or rhoeadine- types as the major alkaloids. PMID- 17401829 TI - [Isolation and chemistry of the alkaloids from plants of the Papaveraceae, LXXXI Glauca - a new section of the genus Papaver]. AB - On the basis of alkaloidal, morphological and chromosomal studies, the section Mecones Bernh. (Syn.: Papaver L., sect. Papaver) has been subdivided into two sections. Thus the new section Glauca J. Novak includes the species Papaver glaucum Boiss. et Hausskn., P. gracile Auch. and P. decaisnei Hochst. et Steud., whereas the section mecones Bernh. only the species P. somniferum L. and P. setigerum Dc. The major alkaloids of the species of the section mecones are the morphinane alkaloids morphine and codeine, whereas those of the section Glauca the rhoeadine and the papaverrubines. The section mecones has the basic chromosome number x = 11, and the section Glauca x = 7. The basic chromosome numbers have been correlated with the morphological features. PMID- 17401830 TI - [On the gas-chromatographical determination of the pyrrolizidine alkaloids of some Senecio-species]. AB - A gas-chromatographical method is described on which seven pyrrolizidine alkaloids (Fuchsisenecionine, Senecionine, Senecivernine, Senkirkine, Retrorsine, O (7)-Angelylheliotridine and Heliotrine) are determinated in a qualitative way and their ratio of concentration in Senecio fuchsii, vernalis, inaequidens and ovirensis, ssp. gaudinii. PMID- 17401831 TI - [Constituents of Aristolochia triangularis]. AB - The roots and stems of Aristolochia triangularis Chamisso, a woody climber growing in Brasil, contain fatty acids as well as (-)-kaur-16-en-18-acid, but no aristolochic acids. The following stigmastane type triterpenes have been isolated: stigmast-4-en-3-on, stigmastan-3,6-dion and stigmast -4-en-3,6-dion as well as kaurane diterpenes: (-)-kaur-16-en, (-), kaur-16-en-18-al, (-)-kauran 16alpha-ol-18-al, (-)-16alpha, 17-epoxykauran, (-)-kaur- 16 - en-18-ol and (-) kaur-15-en-17-ol. In addition sesquiterpenes have been found: (-)-alpha-ylangen, (-)-alpha-copaen, (E)-beta-elemen, gamma-elemen, (E)-(R)-Nerolidol. The lignanes isolated from A. TRIANGULARIS include cubebin, found so far only in Piper cubeba, and galbacin. The new lignans 3,4-dimethoxy-3,4-desmethylendioxycubebin and 3',4' dimethoxy-3',4'-desmethylendioxycubebin have been identified. For better interpretation of the (1)H-NMR-spectra in relation to the stereochemistry, the 6,6'-dinitro derivates are suitable. PMID- 17401832 TI - Secoiridoids and xanthones in the genus Centaurium. AB - Two new acyl secoiridoid glucosides, named decentapicrins A and B, accumulating in the fruits of Centaurium littorale Gilmour spp. littorale, have been isolated by means of preparative column chromatography. Another new compound, named decentapicrin C, could also be detected but in minute amounts and was prepared from decentapicrin B. The close interrelation between these three compounds and the strongly bitter centapicrin (Ia) and desacetylcentapicrin (Ib), accumulating in the fruits of Centaurium erythraea R AFN. [1, 2], have been demonstrated by means of TLC analysis of the products obtained under alkaline treatment. Under these conditions not only hydrolysis, but also acyl migrations, took place. Decentapicrin A is partly converted into desacetylcentapicrin (Ib) and decentapicrin C; decentapicrin B is converted into decentapicrin C. On the basis of this chemical information and spectroscopical data ( (1)H-NMR, (13)C-NMR, UV and IR) the structures of decentapicrins A, B and C are postulated to be Ic, Id and Ie respectively. The single acylation at C (3)-OH and C (4)-OH of the glucose moiety as shown in the structures of decentapicrins A and B is new in iridoid chemistry. Unlike centapicrin (Ia) and desacetylcentapicrin (Ib) the new compounds decentapicrins A, B and C are only weak bitter principles like sweroside (I). PMID- 17401834 TI - A steroid constituent from Solanum abutiloides. AB - A steroidal aglycone, isonuatigenin, has been isolated from the hydrolysate of unripe fruits of Solanum abutiloides. The acetylation of isonuatigenin in weak acid has been studied. PMID- 17401833 TI - Distribution of Cardenolides in Digitalis lanata. AB - By means of a digoxin-RIA it has been shown that all organs and tissues of Digitalis lanata contain cardenolides. PMID- 17401835 TI - The production of acetate derived hydroxyanthraquinones, -dianthrones, naphthalenes and -benzenes in tissue cultures from Rumex alpinus. AB - Tissue cultures set up from roots of Rumex alpinus L. were grown on Murashige and Skoog's medium with kinetin and either 2,4-D or NAA as growth factors. The 2,4-D cultures produced a broader spectrum of hydroxyanthracene, hydroxynaphthalene and hydroxybenzene derivatives than the NAA cultures. In the 2,4-D cultures the anthraquinones chrysophanol, physcion and emodin, the dianthrones of chrysophanol and physcion and their heterodianthrone and the monoglucoside of chrysophanol were identified. Also the naphthaiene-1,8-diols nepodin, nepodinmonoglucoside and methoxynepodin have been identified. The occurrence of 2-acetylorcinol and its monoglucoside was also established. In the NAA cultures accumulation of only chrysophanol, physcion and 2-acetylorcinol could be proved. The yields of these secondary metabolites produced in the NAA cultures were determined to be much lower than those produced by the 2,4-D cultures. PMID- 17401836 TI - Selection of liquid phases for glass capillary gas chromatography of monoterpene hydrocarbons. AB - Glass capillary columns with liquid phases of different polarities were prepared and tested. Of the nine liquid phases used (Apiezon L, SE-30, OV-101, Ucon 50 LB 550X, Emulphor ON-870, Carbowax 20M, FFAP, SP-1000 and Silar 10C) FFAP was the most suitable for the separation of monoterpene hydrocarbons of Scots pine needle oil. For the monoterpene mixtures which contain alpha-thujene and/or tricyclene together with alpha-pinene, only non polar or semipolar columns, such as SE-30, OV-101 or Ucon 50 LB 550X, are to be recommended. Of the polar phases Carbowax 20M and FFAP were found to be the most effective for p-menthadienes. Resolution and relative retention values for some problematic component pairs as well as relative retention times of monoterpene hydrocarbons were calculated. General aspects. the column characteristics, injection technique, effect of temperature on the resolution and artefacts as well as the polarizability of the components as a funktion of the retention have been discussed. PMID- 17401837 TI - Studies on monoterpene glucosides and related natural products. AB - From iridoid-producing cell suspension cultures obtained by selection of callus tissues induced from seedlings of Gardenia jasminoides f. grandiflora, tarennoside (1), a constituent of Tarenna gracilipes, was isolated along with gardenoside (2), geniposide (3) and geniposidic acid (4), constituents of the original Gardenia plant. PMID- 17401838 TI - Iridoid glucosides of Melampyrum laxum. AB - From the aerial part of Malampyrum laxum, Miq. three iridoid glucosides were isolated and identified as melampyroside, mussaenoside and aucubin. PMID- 17401839 TI - [Determination of Structures by H NMR at 400 MHz: two new alkaloids from Tabernaemontana albiflora]. PMID- 17401840 TI - Methylated flavonoid-aglycones from "Matricariae flos". PMID- 17401841 TI - A naturally occurring trans-2-cis-4-isomer of wisanine from Piper guineense. PMID- 17401842 TI - Chemical constituents from Marrubium alysson. PMID- 17401843 TI - Sapogenins from Trigonella polycerata. PMID- 17401844 TI - Gamma-Methyleneglutamic acid in Lilium martagon. PMID- 17401845 TI - Components of Metaplexis japonica Makino. PMID- 17401846 TI - Secoiridoids and Xanthones in the genus Centaurium. AB - Overground parts of Centaurium spicatum (L.) Fritsch (Gentianaceae) from two different habitats have been investigated on their content of Secoiridoid glucosides. Sweroside (I), Swertiamarin (II) and gentiopicrin (III) (= gentiopicroside) have been identified. In plant material of C. spicatum from both habitats sweroside is the major compound. In plant material from one habitat gentiopicrin was found in almost equal amounts as sweroside. Swertiamarin is only present as a minor component. For the isolation of sweroside and gentiopicrin a preparative column chromatographic method using silica gel as adsorbent is described. The chromatographic analysis of these Secoiridoid glucosides has been carried out by means of newly elaborated TLC and HPLC separation systems. The composition of sweroside (I), swertiamarin (II) and gentiopicrin (III) in C. spicatum is compared with that occurring in C. pulchellum (S W.) Druce and C. erytkraea Rafn. (= C. MINUS Moench). On the basis of chemical derivation, chromatographic (TLC and HPLC) and spectroscopical (IR, Mass) data it is shown that sweroside is very likely identical with the compound known as "kantaurin" in literature. PMID- 17401847 TI - [II1: TLC and HPLC-Analysis of Pyrrolizidine-N-oxid Alkaloids of Symphyti Radix]. AB - A new TLC- and HPLC-method for the detection and estimation of the main alkaloids from Symphytum roots, Symphytine, Echimidine, Lycopsamine, and Acetyllycopsamine in their genuine N-oxide form is described. The method provides characteristic finger-prints for the constituents of the Symphytum drugs of different origin and varying harvest seasons. PMID- 17401848 TI - [Isolation of a New Alkaloid (O-Acetylretuline) and a Triterpenoid (Friedelin) from Strychnos henningsii of Zaire]. AB - A new alkaloid (O-acetylretuline) has been isolated from barks and leaves of two samples of Strychnos henningsii collected in Zaire. A triterpenoid (friedelin) is also present in this African species of Strychnos. PMID- 17401849 TI - [A New Antiphlogistic Principle from Sabal serrulata, II]. AB - An acidic polysaccharide from Sabal serrulata Roem. et Schult (Serenoa repens, [Bart] Small) showed at extremely low doses inhibiting activity on the carrageenin paw oedema and pellet test of the rat, but did not influence the proliferative phase of inflammation. PMID- 17401850 TI - Alkaloids from Corydalis marschalliana. AB - The known alkaloids (+/-)-stylopine, (+/-)-sinactine, protopine, (+) bulbocapnine, (-)-domesticine, (+)-corydine and (+)-isoboldine were found in the mixture of alkaloids from Corydalis marschalliana (Papaveraceae), growing in Bulgaria. The alkaloids dehydronantenine, (+)-nantenine and oxonantenine were isolated for the first time from this plant. The spectral data of these alkaloids are given. In addition, three unidentified alkaloids (Cm-4, Cm-11 and Cm-12), in a small quantities, were described. PMID- 17401851 TI - Composition of essential oil of Ledum palustre. AB - The composition of essential oil of Ledum palustre L. was investigated. This oil was obtained by steam distillation of leaf material from plants growing in the Hortus Botanicus at Haren (Groningen). It was separated into seventeen fractions by column chromatography on silicagel and gradient elution with mixtures of petrol (bpt. < 40 degrees C) and diethylether. The investigation was made by GLC and GC-MS. In total, 80 components were identified. About 50 constituents were new, including vitispirane, perillaalcohol, perilla-aldehyde, isopinocamphone, germacrone, etc. Principal components are ledol, palustrol and myrtenal. PMID- 17401853 TI - [Distribution of hydrocyanic Acid in cormophyta: 15.1 new observations on cyanogenesis in Rosaceae]. AB - Prunasin was isolated from leaves and roots of Aruncus silvester (= A. dioicus), leaves of Gillenia trifoliata and seedlings of Kageneckia lanceolata (all Spiraeoideae). Chamaebatiaria millefolium (Spiraeoideae - Sorbarieae) was found to be cyanogenic; in this instance the cyanogenic constituent seems to be different from prunasin, however. Prunasin was isolated also from buds and very young shoots of Crataegus irrasa (Maloideac - Crataegeae). Very young leaves of Crataegus cuneata and C. pedicellata also contain prunasin-like glycosides. Adenostoma fasciculatum, A. sparsifolium, all taxa of Cercocarpus tested and Coleogyne ramosissima were shown to have more or less strongly cyanogenic leaves. Together with Kerria, Neviusia and Rhodotypos these genera deviate from true Rosoideae by being cyanogenic and by having the basic chromosome number 9 instead of 7. The cyanogenic compounds of these taxa seem not to be derived from phenylalanine. Prinsepia uniflora (Prunoideae) has cyanogenic young shoots which do not contain prunasin or a similar compound. Within the species Aruncus silvester, Sorbus aria and Sorbus aucuparia cyanogenesis of leaves was found to be a highly variable character. The taxonomic meaning of cyanophoric compounds in Rosaceae is discussed briefly. PMID- 17401854 TI - Biosynthesis of cucurbitacins in Bryonia dioica seedlings. AB - The biosynthesis of cucurbitacins during the seed germination of Bryonia dioica was studied by analysis of the cucurbitacin-triterpenoid fraction and by tracer experiments with acetate-[2- (14)C]. Isolation of 10alpha-cucurbita-5,24-dien 3beta-ol (9a), the simplest tetracyclic triterpene with a cucurbitane skeleton, supports the view that (9a) is the general precursor of cucurbitacins. Moreover, following the tracer experiments, cucurbitacin E (1a) was the first cucurbitacin formed, whereas the less oxygenated bryodulcosigenin (4a) was not detectable during germination of the plant. In the course of the present investigation, a new pentacyclic triterpene, isomultiflorenol (11a) (possible precursor of bryonolic acid (5a)), was also isolated. PMID- 17401855 TI - [New anthraquinones in Rubia cordifolia roots]. AB - From roots of Rubia cordifolia L. four anthraquinones are isolated: 1-hydroxy 2 methylanthraquinone and nordamnacanthal which have been already isolated in some Rubiaceae, physcion which seems to be new in this family and 1,4-dihydroxy 6 methyl-anthraquinone which seems to be a new quinizarin. PMID- 17401856 TI - Constituents of essential oil of Aframomum giganteum. AB - The composition of the essential oil of Aframomum giganteum was investigated by GC-MS. Forty seven terpenes were identified, the oil contained a high percentage of linalool and linalyl acetate which are mainly responsible for the smell of the A. giganteum stems. Fatty acids, paraffins, n-octanol, n-nonanol and guaiacol were also identified. PMID- 17401857 TI - Chromosomal characteristics and occurrence of main alkaloids in Datura stramonium and Datura wrightii. AB - The population of Datura stramonium L. var. tatula Torr and Datura wrightii Regel was heterogenous in the numerical and structural composition of karyotypes. Datura wrightii Regel contained, as well as aneuploid sets, a 35% karyotype with a diploid set of chromosomes (2n = 24); there were no chromosomes with satellites and there were 1-2 microchromosomes in 29.6% of the metaphases. Datura stramonium L. var. tatula Torr included biotypes with a chromosome number of 21-25; in 88.5% of the metaphases there were chromosomes with satellites and 60% of the metaphases contained 1-3 microchromosomes. According to the content of the main alkaloids Datura stramonium L. var. tatula Torr can be considered a predominantly hyoscyamine type, Datura wrightii Regel also containing scopolamine. PMID- 17401858 TI - [Alkaloids of Pausinystalia macroceras]. AB - A study of the alkaloidal content of trunk-barks of Pausinystalia macroceras (K. Schum.) Pierre, Rubiaceae, resulted in the isolation of six alkaloids, five of which are indole alkaloids that belong to the yohimbane and heteroyohimbane groups; among them, yohimbine was found in major amount. Moreover, the levorotatory isomer of calycanthine, a quinoline dimeric tryptophane derived base, has been isolated for the first time. The phytochemical significance of calycanthine and related alkaloids is discussed. PMID- 17401859 TI - Biogenesis of proteaceous alkaloids. AB - A scheme is presented which outlines the possible mode of biogenesis of alkaloids so far reported from the proteaceous plants Bellendena montana and Agastachys odorata (both from Tasmania), Darlingia darlingiana and D. ferruginea (from Queensland) and Knightia deplanchei and K. strobilina (from New Caledonia). PMID- 17401860 TI - Variability in yield and composition of essential oil in Cymbopogon jawarancusa. AB - Variability in the oil content in natural populations of Cymbopogon jawarancusa is discussed. Races rich in piperitone, phellandrene and other chemical constituents have been identified. The oil producing ability was largely genetic and showed heritability of 79.0 percent. PMID- 17401861 TI - Flavonoids from Chenopodium botrys. AB - From Chenopodium botrys, five flavonoids have been isolated: hispidulin, 1, salvigenin 2, 5-methylsalvigenin, 3, 7-methyleupatulin, 4 and sinensetin, 5. None of them have been previously reported from Ch. botrys. PMID- 17401862 TI - Phytochemical investigation of Cassia multijuga seeds. AB - From the seeds of Cassia multijuga 1,3,8-trihydroxy-2-methyl anthraquinone, 1:3 dihydroxy 6,8-dimethoxy-2-methyl anthraquinone and two glycosides 3-hydroxy-6,8 dimethoxy-2-methyl anthraquinone-1-O-beta- D(+) glucopyranoside and 3-hydroxy 6,8 dimethoxy-2-methyl anthraquinone 1-O-rhamnopyranosyl (1 --> 6) glucopyranoside (rutinoside) have been isolated and their structures elucidated. The last three compounds are new and have not been isolated previously from any plant source. PMID- 17401863 TI - A new flavonol glycoside from seeds of Cassia grandis. AB - A new flavonol glycoside, kaempferol-3-O-beta-D-mannopyranosyl (1-->4)-O-beta-D glucopyranoside has been isolated from the seeds of Cassia grandis L. (N. O. Leguminoseae). PMID- 17401864 TI - Alkaloids of papaveraceous plants. AB - From the whole plants of Corydalis tashiroi Makino a new quaternary protoberberinium base dehydrodiscretamine chloride was isolated along with four known quaternary alkaloids by the application of droplet countercurrent chromatography (DCCC). The structure of the new base was established by spectral and chemical evidence. Eight known tertiary alkaloids were also isolated from the same plant. PMID- 17401865 TI - 13-C NMR data of 3-isoajmalicine and 19-epiajmalicine. PMID- 17401866 TI - Chemical constituents of Lasiosiphon eriocephalus. PMID- 17401867 TI - Isolation of cleomin from roots of Ritchiea longipedicillata. PMID- 17401868 TI - [Pyrrolizidinalkaloids Senecionin and Retrorsin from Senecio inaequidens]. PMID- 17401869 TI - Phytochemical Investigation of the roots of Tacca aspera. PMID- 17401870 TI - Dedifferentiation and loss of Thevetin in Thevetia peruviana callus. PMID- 17401871 TI - Triterpenes of Boswellia frereana. PMID- 17401876 TI - Alkaloid Production in Catharanthus roseus cell cultures VIII. AB - A cell line of Catharanthus roseus (L.) G. Don coded PRL # 200, was characterized with respect to its biosynthetic capabilities for indolealkaloids, in particular catharanthine, in suspension cultures. Other alkaloids isolated are vallesiachotamine isomers, ajmalicine, horhammericine, horhammerinine, vindolinine, 19-epivindolinine and strictosidine lactam. PMID- 17401877 TI - Alkaloids from Strychnos floribunda. AB - From Strychnos floribunda stem bark extracts, the alkaloids bisnordihydrotoxiferine, akagerine, decussine, rouhamine, Strychnocarpine, desacetylisoretuline and isorosibiline were isolated as well as a mixture of the sterols beta-sitosterol, stigmasterol and campesterol. Isorosibiline is a new indole alkaloid derived form desacetylisoretuline, having C-16 configuration inverted to that of rosibiline. PMID- 17401878 TI - [Alkaloids of Annonaceae. XXIX. Alkaloids of Annona muricata]. AB - From leaves, root - and stem - barks of Annona muricata L., seven isoquinoline alkaloids have been isolated: reticuline (main alkaloid), coclaurine, coreximine, atherosperminine, stepharine. Anomurine and anomuricine, two minor alkaloids, are new tetrahydrobenzylisoquinolines, with 5, 6, 7 substituted ring A. The phytochemical significance of these alkaloids is discussed. PMID- 17401879 TI - The thebaine content of ornamental poppies belonging to the papaver section oxytona. AB - Encouraging yields of thebaine have been obtained from Papaver bracteatum grown in Ireland. However climatic conditions adversely affect flower (and subsequent capsule production). Twelve vigorously flowering ornamental cultivars labelled P. orientale or "oriental poppy" were screened for the presence of thebaine. Five of the cultivars had no detectable thebaine. The thebaine content of the remaining 7 cultivars was determined. The capsules of the cultivar "Goliath" had a thebaine content of 3.0%. The identity of the thebaine was confirmed by isolation and examination of spectral evidence. In addition the two known N-oxides of thebaine were isolated and characterised from cv. Goliath. The combination of this chemical evidence and stomatal pore length measurements indicate that the plant labelled P. orientale cv. Goliath is closely related to P. bracteatum Lindl . This cultivar shows promise as a source of thebaine and has a potential in the hybridisation of genuine P. bracteatum to improve its flowering characteristics in mild climates. PMID- 17401880 TI - [Plants from new caledonia: alkaloids of stem-bark of Hernandia cordigera]. AB - The alkaloidal content of of the stem barks of Hernandia cordigera has been studied and compared to that of other Hernandia species. Main alkaloids are phenolic aporphines. PMID- 17401883 TI - Purification and characterization of antiviral substance from the bud of Syzygium aromatica. AB - The antiviral substance from the bud of Syzygium aromatica was purified by ether and ethyl acetate extraction and successive chromatography on Sephadex LH-60, LH 20, G-25 and Lichroprep RP-18. The purified antiviral substance was homogeneous and identified as eugeniin. Eugeniin shows antiviral activity against herpes simplex virus at a concentration of 10 microg/ml. PMID- 17401885 TI - Constituents of the cultivated mulberry tree. AB - From extracts of root bark of the cultivated mulberry tree (a variety of Morus alba L.) three new isoprene-substituted flavanones, Kuwanon D, E and F were isolated. Their structures were shown to be I, II and III, respectively. PMID- 17401886 TI - Studies on coumarins of a chinese drug "Qian-Hu". AB - From the ether-soluble fraction of methanolic extracts of a Chinese Drug "Bai-Hua Quian-Hu", the root of Peucedanum praeruptorum (Umbelliferae), four coumarin compounds, (+/-) praeruptorin A, (+) praeruptorin B (= (+) anomalin), Pd-III and Pd-Ib, were isolated. The chemical structures of Pd-III and Pd-Ib have been established to be (+) 3' ( S)-angeroyloxy-4' ( S)-isovaleryloxy-3', 4' dihydroseselin and (+) 3' ( R)-angeroyloxy-4'-keto-3',4'-dihydroseselin, respectively. (13)C NMR spectra of these compounds are also discussed. PMID- 17401888 TI - Alkaloid Content in Trisomic Mutants of Datura stramonium. PMID- 17401887 TI - Coumarins from Pterocaulon virgatum. PMID- 17401889 TI - [Immunofluorometrische Lokalisation von beta-Glucosidasen in Cicer arietinum L. Keimlingen.]. PMID- 17401890 TI - [Untersuchungen zur genetischen Aktivitat ausgewahlter Cumarinderivate an menschlichen Lymphocyten.]. PMID- 17401891 TI - [Iridoidglucoside der gattung penstemon.]. PMID- 17401892 TI - Variation in the Cardenolide Content of Embryogenic Clumps From Suspension Cultures of Digitalis lanata. PMID- 17401893 TI - [Neuere Erkenntnisse uber Petasites hybridus.]. PMID- 17401894 TI - [Zur Stereochemie der Bisaboloide aus Matricaria chamomilla.]. PMID- 17401895 TI - [Sesquiterpenlactone aus Arnica chamissonis.]. PMID- 17401896 TI - Alkaloids from Rauwolfia salicifolia. PMID- 17401897 TI - [Isolierung und Charakterisierung der blau fluoreszierenden Leitstoffe aus der Rinde von Rhamnus catharticus L.]. PMID- 17401898 TI - Importance of injection techniques in glass capillary gas chromatography. PMID- 17401899 TI - Application of rotation locular counter-current chromatography to the preparative scale separation of natural products. PMID- 17401900 TI - [Polyamid-DC und HPLC von Flavonoid-Aglykonen - ein Vergleich.]. PMID- 17401901 TI - [Qualitatssicherung bei Phytopharmaka.]. PMID- 17401902 TI - Identification of HPLC-Peaks with on-Line Methods (UV, AR, CI-MS) and off-Line Methods (MIR-IR, HPTLC, FD-MS). PMID- 17401903 TI - [Untersuchungen zur Schwankungsbreite des Wirkwertgehaltes in Nerii Oleandri Folium sowie zur Frage: Chemische oder biologische Wirkwertbestimmung?]. PMID- 17401904 TI - [Standardisierung der flavonoide und proanthocyanidine in crataegus drogen und zubereitungen.]. PMID- 17401905 TI - [HPLC-Analyse von Herzglykosiden aus Pflanzenextrakten und Arzneifertigzubereitungen, 2. Mitteilung.]. PMID- 17401906 TI - Physiologal Homeostasis of Essential Oil Production in Lemon Grass (Cymbopogon citratus). PMID- 17401907 TI - [Alkaloide aus Corydalis solida und deren pharmakologische Wirkungen im Vergleich zu Bulbocapnin.]. PMID- 17401908 TI - [Immunologische Eigenschaften der Saponosidfraktion aus Aralia mandshurica.]. PMID- 17401909 TI - Light-mediated control of quinolizidine alkaloid biosynthesis. PMID- 17401910 TI - [Charakterisierung einer UDPG-Isoflavon-7-0-Glucosyltransferase aus Wurzeln der Kichererbse (Cicer arietinum L.).]. PMID- 17401911 TI - [Synthese und Untersuchung substituierter 1-Benzylergometrinderivate.]. PMID- 17401912 TI - [Untersuchungen zur Biosynthese des Vitamin K2. Isolierung des aktivierten Derivates der o-Succinylbenzoesaure aus zellfreien Extrakten von Mycobacterium phlei.]. PMID- 17401913 TI - [Struktur der sapogenine von herba herniariae.]. PMID- 17401914 TI - [Eine einfache Synthese von Dihydroflindersin, Khaplofolin und verwandten Alkaloiden.]. PMID- 17401915 TI - [Phytochemische untersuchungen von saponin-drogen.]. PMID- 17401916 TI - [Sarmentosin-Epoxid, eine neue cyanogene Verbindung aus Sedum cepaea.]. PMID- 17401917 TI - [Ein neues cyanogenes Glucosid aus Acalypha indica.]. PMID- 17401918 TI - Iridoid Glucosides from Euphrasia rostkoviana. PMID- 17401919 TI - Cyanogenesis in Passiflora spp. PMID- 17401920 TI - A New Lignan Glucoside from Euphrasia rostkoviana. PMID- 17401921 TI - Cyanogenic glycosides in butterflies: detection and synthesis of linamarin and lotaustralin in the heliconiinae. PMID- 17401922 TI - Phtalides in the Essential Oil from Roots of Levisticum officinale. PMID- 17401923 TI - [Das Buschmann-Pfeilgift: sekundares Pflanzen- oder Tier-Produkt?]. PMID- 17401924 TI - [Isolierung und quantitative Bestimmung von Phenolglykosiden aus Salix-Species.]. PMID- 17401925 TI - Analysis of Pyrrolizidine Alkaloids from the Symphytum officinale Species Complex and S. asperum. PMID- 17401927 TI - [Praparative Trennung von Furanocoumarinen mittels zentrifugaler Dunnschichtchromatographie.]. PMID- 17401926 TI - Separation and Determination of Phenol Glycosides with anti-Pseudomonas Activity from Poplar leaves by Droplet Counter Current Chromatography (DCCC). PMID- 17401928 TI - [Hochdruckflussigchromatographische Bestimmung der Cyanglykoside Cardiospermin und Cardiosperminsulfat mittels Ionenpaarchromatographie.]. PMID- 17401929 TI - [Analyse qualitative et quantitative des terpenes de Ginkgo biloba par HPLC.]. PMID- 17401930 TI - Determination of hyoscyamine levels in plant material by radioimmunoassay. PMID- 17401931 TI - Evaluation of Coumarins and Dicycloethers in Matricaria chamomilla Flowers and Chamomile Extracts by Reverse-Phase HPLC. PMID- 17401932 TI - [Analytische und pharmakologische Aspekte des atherischen Oles von Valeriana officinalis.]. PMID- 17401933 TI - Valepotriate content of different valerianaceae callus- and suspension cultures. PMID- 17401934 TI - [Zur qualitat von pfefferminz- und kamillen-teedroge aus dem handel.]. PMID- 17401935 TI - [Analytik der Phenylpropylamine von Catha edulis mittels HPLC.]. PMID- 17401936 TI - Degradation of Gramine by Cell Suspension Cultures of Barley (Hordeum vulgare). PMID- 17401937 TI - [Das atherische Ol der Fruchte von Ammi visnaga.]. PMID- 17401939 TI - Alkaloids from Strempeliopsis strempelioides. PMID- 17401938 TI - Alkaloids from Corydalis cava. PMID- 17401940 TI - Decussine, a New Muscle-Relaxant Alkaloid from Strychnos decussata. PMID- 17401941 TI - The Isolation of Colchicoside, Colchicine and Oil from Colchicum autumnale seed. PMID- 17401942 TI - Action of Phytohormones and Fungal Infections on the Content of Antimicrobial Acridone Alkaloid Epoxides in Callus Cultures of Ruta graveolens L. PMID- 17401943 TI - [Sanguinarin- und protopinalkaloide in papaver-suspensionskulturen.]. PMID- 17401944 TI - Isolation of dill seed oil by hydrodistillation following solvent extraction. PMID- 17401945 TI - Fungitoxic activity of the secoiridoid glucoside gentiopicrin (gentiopicroside). PMID- 17401946 TI - Thin-layer and gaschromatographic separation of monoterpenes from different essential oils of plants. PMID- 17401947 TI - Antifungal Activity of Alpinia glanga. PMID- 17401948 TI - Malindine, a New Quaternary Indole Alkaloid, Isolated from Strychnos decussata. PMID- 17401949 TI - Two New Orange Coloured Alkaloids from Pycnarrhena longifolia. PMID- 17401950 TI - Pharmacognostical Studies on Synclisia scabrida. PMID- 17401951 TI - Tertiary Alkaloids of Strychnos soubrensis. PMID- 17401952 TI - Pharmacognostical Researches for the Therapeutic Use of Genista tinctoria. PMID- 17401953 TI - Neue, lipophile Benzofuranderivate aus Ratanhiawurzel. AB - Two phenols have been isolated from the petroleum-ether extract of Ratanhiae radix Ph. Eur.; structure elucidation has been made by means of the following chemico-physical methods: UV, IR, (1)H-NMR, (13)C-NMR-spectroscopy and mass spectrometry, microhydrogenation and thermofractography. Structures are: 2-(p methoxy-o-hydroxyphenyl) -5-propenyl (trans)-benzofurane; 2-(p-hydroxyphenyl)-3 methyl-5-propenyl-(trans)-benzofurane. Both compounds - newly discovered in Ratanhiae radix - proved to be effective UV-light filters. They could be useful in sun protection preparations. PMID- 17401954 TI - [In Vitro-Untersuchungen uber die expektorierende Wirkung der Phenylpropanderivate der Haselwurz.]. PMID- 17401955 TI - Biotransformation of cardenolides by plant cell cultures1. AB - A culture of DAUCUS CAROTA cell line Ca68, previously shown to specifically hydroxylate digitoxigenin at the C-5 position, has been shown also to convert gitoxigenin and oleandrigenin to their corresponding 5beta-hydroxy derivatives. These two products have not been identified previously. 5beta-Hydroxygitoxigenin has been identified by physico-chemical methods and data on its mass spectrum and (13)C-NMR spectrum are reported. 5beta-Hydroxyoleandrigenin was identified from its high resolution mass spectrum. Addition of gitoxigenin at 72 hrs after inoculation resulted in a two fold increase in the rate of bioconversion compared to addition at 0 hr. The results are discussed in relation to other biotransformation reactions and with respect to improving the yields of plant products and the formation of novel compounds. PMID- 17401956 TI - Phosphate Mediated Regulation of Cinnamoyl Putrescine Biosynthesis in Cell Suspension Cultures of Nicotiana tabacum. AB - The accumulation of cinnamoyl putrescines by cell suspension cultures of NICOTIANA TABACUM was enhanced manifold by phosphate limitation of the culture medium while growth was reduced under such conditions. The enhanced product formation was preceded by a large increase and subsequent decline of phenylalanine ammonia lyase activity and a smaller increase of the activity of ornithine decarboxylase. Phosphate concentrations commonly used in cell culture media suppressed the product accumulation. PMID- 17401957 TI - Selection of tobacco cell lines with high yields of cinnamoyl putrescines. AB - Cell lines of NICOTIANA TABACUM resistant to either L-phenylalanine, DL-meta fluorophenylalanine (MFP) or L-ornithine, have been selected in order to establish by this means cell strains with higher yields of cinnamoyl putrescines. Except for three MFP-resistant cell lines, all selected strains accumulated 2 to 5 times more cinnamoyl putrescines than the non-resistant wild type cells. In some lines this increase was accompanied by increased activities of enzymes which were likewise involved in the biosynthesis of cinnamoyl putrescines and in the metabolism/degradation of the selecting agents. The qualitative pattern of phenolic compounds did not change in any of the resistant cell lines. PMID- 17401958 TI - Seasonal variation of saponins, sucurose and monosaccharides in cultivated ginseng roots. AB - Yield of methanolic extract of fresh roots of Ginseng harvested in winter was found to be more than two-fold grater than from roots collected in summer; this remarkable increase is mainly due to the large increase of sucrose in roots in winter. On the other hand, biologically active dammarane-saponins in the roots increase in summer. These results indicate that roots should be harvested in summer for the production of high quality Ginseng extracts. PMID- 17401959 TI - NMR spectroscopy of naturally occurring phloroglucinol derivatives. AB - Carbon-13 NMR spectra of eight 2-acyl- and -6-methyl-substituted filicinic acid (4,4-dimethyl-1,3,5-cyclohexantrione) derivatives were recorded in hexadeuterioacetone and in deuteriochloroform and the signals assigned on basis of chemical shifts and J (CH) coupling considerations. The data prove that the prevailing tautomeric structure of these derivatives is the monoketonic one with the carbonyl function in position 3 and hydroxyl groups in positions 1 and 5, the former being hydrogen bonded to the acyl side chain carbonyl. The 2-acyl-6,6 dimethyl compounds have the diketonic structure and a hydrogen-bonded hydroxyl group in position 3. The skeletal structure of 2-butyryl-4-hydroxy-4,6-dimethyl 1,3,5-cyclohexantrione is identical with that of humulone. This compound and its homologues also exist in the monoketonic tautomeric form, where C-3 carries the carbonyl function. In the solvents used the 6-methyl-2,3-dihydropyrane-2,4-dione exists predominantly as a single tautomer having two ring double bonds; the remaining carbonyl group is a part of an alpha, beta-unsaturated lactone grouping. PMID- 17401960 TI - Constituents of Pygeum africanum Bark. PMID- 17401961 TI - Mulberrofuran B, a new isoprenoid 2-arylbenzofuran from the root bark of the cultivated mulberry tree. PMID- 17401962 TI - Flavonoid Constituents of Aegialophila pumila. PMID- 17401963 TI - Pyrrolizidine Alkaloids from Ligularia japonica. PMID- 17401967 TI - The content of Psilocybin in Norwegian Psilocybe semilanceata. AB - Psilocybe semilanceata contains hallucinogenic indole alkaloids and has been used as a narcotic drug in Norway. The content of psilocybin, the major constituent in Norwegian P. semilanceata, has been investigated as well as its habitats and its distribution throughout the country. The mushroom is growing on grassy sites in most parts of Norway from the middle of August to the middle of October. The psilocybin content in dried mushrooms varied considerably, from 0.17 to 1.96%. The highest concentrations in percent was found among the smaller mushrooms whereas the content in mg was highest in larger mushrooms. The content of psilocybin makes P. semilanceata to a potent hallucinogenic drug. PMID- 17401968 TI - Biotransformation of monoterpenes by mentha cell lines: conversion of menthone to neomenthol. AB - Following previous results, which indicated that cell lines derived from different Mentha chemotypes were either capable or not capable to biotransform pulegone into isomenthone, we studied menthone biotransformation by in vitro cultured Mentha cell lines. All the six cell lines did transform (-)-menthone into another monoterpene. The latter was identified by GLC, TLC and NMR techniques as (+)-neomenthol. None of these cell lines reduced (+)-isomenthoneto the corresponding alcohol. These results indicate a stereospecificity in respect to both precursor and product in this plant cell biotransformation system. PMID- 17401969 TI - Radioimmunoassay of digoxigenin glycosides in digitalis lanata. AB - Radioimmunoassay (RIA) has been used in the analysis of digoxigenin glycoside contents of crude ethanolic extracts prepared from fresh leaf samples of Digitalis lanata Ehrh. The commercial antidigoxin antiserum used in the assay was highly specific against the digoxigenin aglycone. Steroid derivatives with only slight structural differences from digoxigenin remained practically unassayed. The method is highly sensitive, and excellent values for accuracy, precision and correlation were obtained. Digoxigenin glycoside levels of second-year D. lanata leaves were found to be considerably higher than those of frist-year leaves. PMID- 17401970 TI - Assay of volatile oil-cyclodextrin complexes by pyrolyzis gas-chromatography. AB - A pyrolytic gaschromatographic process was developed for the routine assay of both cyclodextrin inclusion complexes, and for the differentiation of the mechanical mixture of the same oil with beta-cyclodextrin. The method is a direct assay of low material demand. PMID- 17401971 TI - [Iridoids of Borreria verticillata]. AB - Seven iridoids are isolated from Borreria verticillata root barks: daphylloside 1, asperuloside 2, feretoside 3, methyl desacetylasperulosidate 4, desacetylasperuloside 5, asperulosidic acid 6 and desacetylasperulosidic acid 7. Their structures result from spectroscopic analysis and are proved by chemical correlation. PMID- 17401972 TI - [Structure of the Oligosaccharide Chain of Chrysantellin A by (13)C-NMR]. PMID- 17401973 TI - Structure of methyl adenophorate and triphyllol, triterpenoids of Adenophora triphylla var. japonica roots. AB - From the crude drug "shajin", the roots of Adenophora triphylla var. japonica, two novel triterpenoids, methyl adenophorate and triphyllol, have been isolated and these structures have been determined as those represented by formula I and II, respectively, on the basis of chemical and physical evidence. PMID- 17401974 TI - Constituents of West African medicinal plants. AB - Extracts of the stems and roots of Heptacyclum zenkeri Engl. (Menispermaceae) (syn. Penianthus Zenkeri Diels) collected in Ghana afforded the protoberberine alkaloids palmatine and jatrorrhizine and the aporphine alkaloid magnoflorine. PMID- 17401975 TI - Justicisaponin-I, a new triterpenoid saponin from Justicia simplex. AB - A new triterpenoid saponin, named justicisaponin-I, has been isolated from Justicia simplex D. Don. (Acanthaceae). It has been assigned oleanolic acid-3-O beta-D-glucopyranosyl-4'-O-ferulate structure (1) on the basis of chemical transformation and comprehensive spectral evidence. Justicisaponin-I has produced significant sperm acrosomal membrane stabilizing action and may prove to be a potent antifertility agent. PMID- 17401976 TI - [Flavones and Sesquiterpene Lactones of Volutarella divaricata]. AB - A flavone, jaceosidine, and a sesquiterpene lactone, cynaropicrine, have been isolated from the aerial parts and flowers of Volutarella divaricata. The structures were established by chemical (acetylation, saponification, formylation) and spectroscopic methods (I.R., U.V., R.M.N., Mass spectra). This is the first report of these products from this plant. PMID- 17401977 TI - Reversed-phase high-performance liquid chromatography analysis of apigenin and its glucosides in flowers of Matricaria chamomilla and chamomile extracts. AB - Quantitative estimation of the flavones apigenin, apigenin-7-glucoside and apigenin-7-acetylglucoside in ligulate florets of Matricaria chamomilla was performed by HPLC using a reverse-phase column and eluting with acetonitrile/water, acetic acid system. These flavonoids were detected at 335 nm. Apigenin and its glucosides were not found in the tubular florets. The method was also applied to the estimation of these flavonoids in chamomile extracts. PMID- 17401978 TI - Isolation of four ecdysones from Ajuga iva roots and a rapid semiquantitative method for ecdysone determination. AB - The ethereal extract of AJUGA IVA (L.) S CHREB. roots, yielded 4 ecdysones. Three were proved to be cyasterone, makisterone A and ecdysterone, by using UV, IR, MS spectral methods and comparison with standard samples. Spectral data indicate that the fourth ecdysone is similar to cyasterone but having an additional hydroxy group in the side-chain. The chromatographic pattern of ecdysone content in roots, stems and leaves is presented as well as a rapid and simple Semiquantitative method for their estimation. The roots had the highest total content (0.381%). PMID- 17401979 TI - Flavonoids from the flowers of Clerodendron infortunatum. AB - Apigenin, acacetin and a new flavone glycoside, characterised as the methyl ester of acacetin-7-0-glucuronide have been isolated from the flowers of Clerodendron infortunatum. PMID- 17401982 TI - Effect of seed number and fruit weight on content and localization of glycoalkaloids in Solanum khasianum. PMID- 17401983 TI - [Composition of the essential oil of Persea americana]. PMID- 17401984 TI - Isolation of 2-hydroxy-3-butenyl thiocyanate, epigoitrin, and adenosine from 'Banlangen', Isatis indigotica root. PMID- 17401988 TI - Sterols of Delphinium ajacis; production and metabolic relationships in whole plants and callus tissue. AB - Sterols from whole nonsterile Delphinium ajacis plants and from sterile tissue cultures (callus) were identified and determined quantitatively. The major sterols in the whole plant tissues were sitosterol, campesterol and stigmasterol, whereas those in the callus tissue were stigmastanol, 24-ethylidenelophenol and Delta (7)-stigmastanol. Of the 21 compounds identified in callus tissue, 5 were not present in the whole plant, most notably Delta (7)-stigmastanol. For both sources of tissue, the sterol predominating in one was a minor component in the other (whole plant/tissue cultures: sitosterol 57%/5%; stigmastanol 2%/35%). On a tissue dry-weight basis, the amount of sterols isolated from callus tissue culture was ten to twenty times that obtained from the whole plant. Qualitatively the sterols from both sources fit into a metabolic scheme which proceeds from cycloartenol through 4,4-dimethylsterols and 4-methylsterols to sterols. A proposed metabolic pathway shows the differences in accumulation of sterols in the two types of tissue. The increase in sterol production in cultured cells, especially when favored by growth conditions, has promise for industrial application and in organic synthesis. PMID- 17401990 TI - [About major alkaloids of Strychnos scheffleri from Zaire]. AB - Leaves and stem barks of S. scheffleri Gilg afforded eight dihydroindole alkaloids. Four tertiary monomers (O-methyl Na-acetylstrychnosplendine, Na acetylstrychnosplendine, strychnobrasiline and strychnofendlerine) have been isolated from leaf material while stem barks yielded one tertiary monomer (desacetylisoretuline), one tertiary symmetrical dimer (bis-nordihydrotoxiferine) and two quaternary monomers (mavacurine and fluorocurine). PMID- 17401991 TI - [The major quaternary alkaloids of Strychnos variabilis from Zaire]. AB - Two quaternary alkaloids have been isolated from the root barks of Strychnos variabilis collected in Zaire. The structure of these alkaloids (mavacurine and fluorocurine) could be deduced from spectral data: UV, MS, IR and (1)H NMR. Moreover, the stereochemistry could be assigned by (1)H NMR at 360 MHz. PMID- 17401992 TI - Alkaloids from roots of Aconitum crassicaule. AB - From the roots of Aconitum crassicaule, four diterpenoid alkaloids were isolated. Two of these were identified as chasmanine (5) and yunaconitine (4). The other two, crassicauline A and crassicauline B, are new alkaloids. The structure of crassicauline A, C (35)H (49)O (10)N, has been elucidated as structure (1) on the basis of chemical and spectral data, while the structure of crassicauline B, C (27)H (31)O (4)N, is still to be determined. PMID- 17401993 TI - High-level of cyclic AMP in the jujube plum. AB - A large amount of cyclic AMP-like substance was found in the imported commercial jujube plum, Zizyphi Fructus, commonly used in traditional Chinese Medicine. The fresh fruits of the jujube plum, as obtained from a local botanical garden, also contained comparable amounts of the substance. The substance was purified to an almost pure state from commercial material and was identified as cyclic AMP (adenosine-3',5'-monophosphate) by both chemical and biochemical means. Thus, it has been established that cyclic AMP content of the fruits of jujube plum reaches up to 500 nmoles/g dry weight; which is more than 10 times higher than the maximum value so far reported in other plants. PMID- 17401994 TI - Composition of essential oils of some senecio species. AB - The composition of the essential oils of Senecio vulgaris, S. jacobaea, S. congestus and S. fuchsii was investigated by means of GLC, CC and GC-MS. The principal constituents differ; S. congestus: 1-tridecene and 1-undecene, S. jacobaea: germacrene D and 1-undecene, S. vulgaris: beta-caryophyllene, alpha copaene and myrcene, S. fuchsii: anhydro-oplopanone and an unknown sesquiterpene C (15)H (24). In all the oils anhydro-oplopanone is present. The spectra of this compound and of the synthesized anhydro-oplopanols are given. PMID- 17401995 TI - Polyploid complexes within the genus Galium. AB - Anthraquinones of the shikimate-glutarate-mevalonate pathway occur generally in the roots of Galium species. In order to use this chemical character of the genus Galium on the level of polyploid complexes, a chemical analysis has been carried out of anthraquinones which accumulate in the roots of cytologically defined Galium album Mill. (2n = 44). Fractionation of anthraquinone mixtures was performed by applying successive extractions with solvents of the eluotropic sequence and aqueous alkaline solutions of increasing pH. Single anthraquinones were obtained by means of preparative thin-layer chromatography. The identification of ten isolated compounds is described on the basis of chromatographic, chemical and spectroscopical data. Four of the described anthraquinones are new structures. PMID- 17401996 TI - [New furoquinolins from Monnieria trifolia]. AB - Four new furoquinoline alkaloids (evolatine, evoxine, 6-methoxy-7-hydroxy dictamnine, haplopine) are isolated from the leaves of Monnieria trifolia. The structures are elucidated by spectrometric methods and some chemical transformations. PMID- 17401997 TI - Methoxylated flavonols and flavanones from Eupatorium odoratum. AB - From the NaOH-soluble fraction of the C (6)H (6) extract of Eupatorium odoratum growing in Nigeria, four known methoxylated flavonols and flavanones were isolated and identified as isosakuranetin, kaempferide, sakuranetin and tamarixetin. A discussion of the flavonoid pattern in the hitherto studied Eupatorium species is also presented. PMID- 17401998 TI - Further study on dammarane-saponins of leaves and stems of American Ginseng, Panax quinquefolium. PMID- 17401999 TI - Dynamics of solasodine accumulation in developing berries of Solanum sysimbrifolium. PMID- 17402001 TI - Naphthalene glycosides in Cassia senna and Cassia angustifolia. AB - From leaves and pods of Cassia senna L. and C. angustifolia Vahl. were isolated the naphthalene glycosides 6-hydroxymusizin glycoside and the new tinnevellin glycoside. The structures were established mainly by spectroscopic methods ( (1)H NMR, (13)C NMR, MS). PMID- 17402003 TI - Quinovic acid glycosides from roots of Macfadyena unguis-cati. AB - Two glycosides, isolated in good yields from roots of Macfadyena unguis-cati (L.) A. G entry (syn. Doxantha unguis-cati (L.) Miers), gave on hydrolysis the same aglycone (quinovic acid). The sugar moieties have been identified as fucose and glucose respectively. This is the first time that saponins have been found in Bignoniaceae and their occurrence may account of the use of the plant in folk medicine. PMID- 17402004 TI - Three new iridoid glucosides from Hedyotis diffusa. AB - Three new iridoid glucosides, 6-O-p-coumaroyl, 6-O-p-methoxycinnamoyl and 6-O feruloyl ester of scandoside methyl ester, have been isolated along with a known glucoside, asperuloside from the aerial parts of Hedyotis diffusa Willd. PMID- 17402005 TI - [Iso-Scrophularioside, a new iridoid glucoside from Penstemon eriantherus]. AB - From the aerial parts of Penstemon eriantherus Pursh a new iridoid glucoside was isolated and structurally elucidated as 10-cinnamoylaucubin mainly by spectroscopic methods ( (1)-NMR, (13)-NMR, MS). PMID- 17402006 TI - Investigation of Pinus mugo essential oil oxygenated fraction by combined use of gas chromatography and dry column chromatography. AB - The oxygenated compounds of Pinus mugo Turra essential oil were investigated by a combination of GC and dry column chromatography (DCC) coordinated by GC data processing. The collected data resulted in a bar graph ("normalized" gas chromatogram) giving the RRT's and relative amounts of 68 components; 38 of them were identified by MS and IR. The described procedure may be used for essential oil analysis in general. PMID- 17402007 TI - Confirmation of structures of semecarpus biflavanones A1 and A2. AB - The biflavanone mixture from semecarpus anacardium L. was reisolated and dehydrogenated with I (2)/KOAc in AcOH to yield the corresponding, relatively more stable, biflavone mixture which was methylated using diazomethane. Two biflavone methyl ethers SA1 and SA2 were obtained which confirmed the structures proposed for the parent biflavanones A1 and A2 respectively. PMID- 17402008 TI - Alkaloids from Corydalis bulbosa. AB - A new 4,5-dioxoaporphine, named corydione, was isolated from the whole herb of Corydalis bulbosa (L.) Dc (Papaveraceae), together with dehydronantenine, (-) stylopine, (+)-bicuculline, (-)-adlumidine, (+)-nantenine, (+)-bulbocapnine, protopine (-)-domesticine, (+)-corydine, predicentrine, oxonantenine, (+) isoboldine and nandazurine. Glaucine and thaliporphine were detected by means of thin-layer chromatography. In addition, one minor alkaloid Cb-4, uncompletely identified, was described. The spectral data of corydione, predicentrine and nandazurine are given. PMID- 17402012 TI - Genetic improvement of Papaver bracteatum: heritability and selection response of thebaine and Seed Yields. AB - The heritability and repeatability of various characters affecting thebaine and seed yields in Papaver bracteatum were estimated from parents and progeny families during two consecutive years of growth. Thebaine and seed yields per plant showed the lowest heritability and therefore low selection response. High selection responses were obtained for thebaine content of the capsules and mean weight per capsule, and are therefore the most appropriate criteria for increasing the thebaine yield per plant or per unit area. Similarly seed yield per capsule is more advantageous as a selection criterion for increasing the yield of seeds. PMID- 17402013 TI - A simple method for purification of glycoalkaloids in the quantitative estimation of Solanum khasianum. AB - The property of Hydroxylapatite (Ca (5) (PO (4)) (3) OH) to adsorb plant components selectively is used to isolate and purify steroid glycoalkaloids from Solanum khasianum. This method is suitable for botanists seeking higher alkaloid yielding plants and can be performed in experimental plantations where limited laboratory facilities are available. PMID- 17402014 TI - Dihydroflavanonols from Cedrus deodara, A (13)C NMR study. AB - High resolution (13)C NMR study of taxifolin, cedeodarin, cedrin and their methyl ethers allowed unambiguous placement of the Me in 5,7-dihydroxyflavanonol nucleus, besides providing other valuable information on the substitution pattern in the molecule. PMID- 17402015 TI - New phenolic components of Didymocarpus pedicellata. AB - A chemical investigation of Didymocarpus pedicellata has resulted in the isolation of 8-hydroxy-5, 6, 7-trimethoxy-flavanone, 2'-hydroxy-4', 5', 6' trimethoxychalcone, 5, 6, 7, 8-tetramethoxy flavanone, pedicellin, methyl pedicin and beta-sitosterol. 8-Hydroxy-5, 6, 7-trimethoxy flavanone and 2'-hydroxy-4', 5', 6'-trimethoxy chalcone are hitherto new natural products while 5, 6, 7, 8 tetramethoxy flavanone has been isolated from D. pedicellata for the first time. PMID- 17402016 TI - Triterpene quinone-methides from Gymnosporia montana. AB - Iguesterin, pristimerin, tingenone, beta-amyrin, beta-sitosterol and maytenonic acid have been isolated from the light petroleum extract of the stem bark and iguesterin, pristimerin, tingenone, beta-amyrin and beta-sitosterol from the root bark of Gymnosporia montana. PMID- 17402017 TI - Isolation of bergapten and marmesin from Balanites aegyptiaca. PMID- 17402018 TI - Root wood alkaloids of Rauwolfia macrophylla. PMID- 17402019 TI - Flavonoids of Neurolaena venturana. PMID- 17402020 TI - Phytochemical investigation of Sedum acre L. PMID- 17402021 TI - [Tussilagine - a new pyrrolizidine alkaloid from Tussilago farfara]. PMID- 17402022 TI - Constituents of Aegle marmelos. PMID- 17402024 TI - [Structure of silandrin and silymonin, two new flavanolignans from a white blooming Silybum marianum variety]. AB - The structures of two flavanolignans, (-) silandrin and (+) silymonin, from a white flowering variety of Silybum marianum, have been elucidated as 3-desoxy isosilybin and 3-desoxy-silydianin partly by spectroscopic investigations, partly by chemical transformation into other derivatives and their partial synthesis. In addition 5,7-dihydroxy-chromone has been isolated as a new compound. PMID- 17402025 TI - [To the classification of commercial Calamus drugs]. AB - Acorus calamus L. yields different races depending upon the number of chromosomes in the plant cells and consequently different drug types with varying content of beta-asarone. The phenylpropane derivative proved to be carcinogenic in several animal tests. The diploid drug does not contain traceable amounts of beta asarone, the triploid variety contains 0.3 per cent in the rhicome, the tetraploid yielded two races with a) 2 per cent and b) between 4 and 8 per cent beta-asarone. For reasons of optimal drug safety the use of asarone-rich races should be avoided. They can be distinguished by a fast and safe method of determining the extinction of the drug extract at the beta-asarone maxima of 253 and 303 nm. The determination of beta-asarone is possible using quantitative direct evaluation of thin-layer chromatograms on alumina layers with an efficient separation of the asarone. A good selectivity, obtainable at 300 nm, allows determination of beta-asarone down to 0.1 microg in drug extracts. PMID- 17402026 TI - [Rubia cordifolia roots. II: new quinones]. AB - From roots of Rubia cordifolia L. two anthraquinones are isolated: 1,4-dihydroxy 2-methylanthraquinone and 1,5-dihydroxy 2-methylanthraquinone and 3-prenyl methoxy 1,4-naphthoquinone. PMID- 17402027 TI - Alkaloid, lignan and flavonoid constituents of Haplophyllum tuberculatum from Sudan. AB - The aerial parts of Haplophyllum tuberculatum collected in Sudan have yielded the furoquinoline alkaloid skimmianine, the lignan justicidin-A, and 5,7,4' trihydroxy-6-methoxy-3-O-glucosyl flavone. Skimmianine was identified by direct comparison with authentic material; justicidin-A by analysis of spectral data and by examination of lanthanide induced shifts in the PMR spectrum; and 5,7,4' trihydroxy-6-methoxy-3-O-glucosyl flavone by spectral analysis of the original isolate and of its hydrolysis product. PMID- 17402028 TI - Transformation of alkaloids of the Colchicine type in leaves and flowers of Colchicum autumnale and C. byzantinum. AB - It was found that the so far described variations in the content of colchicine alkaloids in leaves and flowers of Colchicum autumnale L. and C. byzantinum Ten. are due to enzymatic O-demethylation and degradation of colchicine (I), N deacetyl-N-formylcolchicine (XI), and demecolcine (XVII) during drying and storing of the plant material. A simplified method was elaborated to separate the neutral, phenolic, basic, and basic-phenolic fractions of tropolone alkaloids. PMID- 17402031 TI - Tetraphyllin B from Turnera diffusa. AB - Tetraphyllin B has been isolated from the tropical American species Turnera diffusa Willd. The structure was established by NMR and MS. PMID- 17402032 TI - HPCL Separation and quantitative determination of valepotriates from Valeriana kilimandascharica. AB - Valepotriates, mainly isovaltrate and valtrate, have been separated and quantitatively estimated by reversed-phase HPLC in the leaves, flowers, stems and rhizomes of VALERIANA KILIMANDASCHARICA. The isovaltrate/valtrate concentration reaches a maximum of 5.89% in the leaves, 3.84% in the flowers, 3.17% in the stems and 5.15% in the rhizomes. A micro Bondapak C (18) column using MeOH-H (2)O mixtures as eluant is suitable for a baseline separation of isovaltrate, valtrate, acevaltrate and baldrinal at UV 254 nm in 15 min and didrovaltrate and IVHD-valtrat at UV 208 nm in 10 min. Relative standard deviation for quantitative determinations is approximately 1.5% for valepotriate contents of 1%. This method is adaptable for routine analysis of crude extracts. PMID- 17402036 TI - Constituents of the Trunk Resin of Eperua purpurea. AB - The major diterpene acids and neutral constituents of Eperua purpurea Benth. have been isolated and identified. The first isolation of crystalline racemic labda-8 (20), 13-dien-15-oic acid and of a high molecular weight (580) ester derived from labd-8 (20)-en-15-oic acid and the corresponding alcohol, is described. PMID- 17402042 TI - [Qualitative und quantitative Analyse von Herzglykosiddrogen durch HPLC Verfahren]. AB - The qualitative and quantitative estimation of cardiac glycosides from the leaves of Nerium species by an HPLC-method is described. The analysis is performed on a C 18 reversed phase column by gradient elution. For the quantitative estimation of the cardenolides Oleandrin is used as external standard. The cardenolide content in Nerium leaves of different origin showed marked differences in the quantitative distribution among the major glycosides. PMID- 17402043 TI - Alkaloids from Turkish samples of Papaver orientale and P. pseudo-orientale. AB - The alkaloids obtained from the capsules of five different collections of wild P. orientale and sixteen different collections of wild P. pseudo-orientale have been investigated and the chromosome numbers of some of the samples determined. Oripavine was the major alkaloid obtained from four of the P. orientale collections, two of which had a diploid chromosome number of 28. These four samples contained isothebaine and alpinigenine as minor alkaloids whereas two contained mecambridine and orientalidine and the other two contained thebaine and salutaridine. The remaining sample of P. orientale contained mecambridine as the major alkaloid and also some orientalidine. Thirteen of the sixteen samples of P. pseudo-orientale contained isothebaine, mecambridine and orientalidine as their major alkaloids with thebaine and salutaridine as minor alkaloids. Alpinigenine was detected in three samples. The diploid chromosome number of four of the thirteen samples was determined as 42. Two samples of P. pseudo-orientale contained salutaridine and thebaine as major alkaloids and had a diploid chromosome number of 14 whereas the remaining sample yielded salutaridine as the major alkaloid with isothebaine, mecambridine and orientalidine as minor alkaloids and a diploid chromosome number of 28. PMID- 17402044 TI - [Biosynthesis of Capsaicinoids in Capsicum annuum L. var. annuum:]. AB - The biosynthesis of the branched fatty acid moiety of Capsaicinoids was examined by radioisotopic technique. Labelled nordihydrocapsaicin (NDHC), homocapsaicin I (HC I) and homodihydrocapsaicin I (HDHC I) were isolated from CAPSICUM fruits which had been fed L-leucine-U- (14)C via cotton wicks inserted into the stems. After the administration of L-i-leucine-U- (14)C radioactivity could be detected in the HC II- and HDHC II-fractions. As L-valine serves as precursor of L leucine, not only labelled capsaicin (C) and its dihydroderivative (DHC) resulted when L-valine-U- (14)C was fed, but also NDHC, HC I and HDHC I. The results demonstrate that valine, i-leucine and leucine are direct precursors of the respective even- and odd-numbered branched fatty acid moieties of the capsaicinoids. PMID- 17402045 TI - [Isolation of solasonine from glucoalkaloid mixtures by fungal fermentation.]. AB - A strain of Aspergillus niger grown on a medium containing 60% of Solamargine and 40% of Solasonine was able to hydrolyse selectively Solamargine. It was thus possible to extract pure Solasonine. Different parameters influencing the speed of hydrolysis of the Solamargine were examined. PMID- 17402046 TI - [Inhaltsstoffe aus Wurzelrinde und Stammholz von Afraegle paniculata]. AB - The isolation of the coumarins xanthyletin, imperatorin, (+)-heraclenin and xanthoxyletin from the rootbark of A. Paniculata (Schum. et Thonn.) Engl. is reported. From the heartwood of the same plant the coumarins scoparon, xanthotoxin, imperatorin, (+)-heraclenin, 6-hydroxy-7-methoxy-coumarin and marmesin as well as the alkaloids dictamnine, haplopine and atanine have been isolated and identified. PMID- 17402047 TI - Alkaloids from Senecio aegyptius and S. desfontainei. AB - Otosenine and senecionine/seneciphyllme were isolated from Senecio aegyptius and S. desfontainei, respectively. Senecionine and seneciphylline easily cocrystallize; m.p. and IR-spectra of mixtures of both alkaloids and their separation by PC are described. IR evidence of individual alkaloids and of the alkaloid-pair are shown. PMID- 17402048 TI - Structure of cadabalone. AB - From the pods of Cadaba fructicosa L., a new dilactone designated as cadabalone, has been isolated. Based on reactions and spectral data, structure I has been assigned for cadabalone. PMID- 17402049 TI - [Quantitative TLC analysis of Valepotriates.]. PMID- 17402050 TI - [Alkaloids of Borreria capitata.]. PMID- 17402051 TI - Candicine from Stapelia gigantea. AB - Hordenine (N, N-dimethyltyramine) was found previously in the title plant. The detection of tyramine, N-methyltyramine, and choline and the crystallization of candicine (N, N, N-trimethyltyramine chloride) is now reported for this species. PMID- 17402052 TI - A new Dihydrofurocoumarin from Apium graveolens. PMID- 17402053 TI - Isoboldine and lupenone from Neolitsea fuscata. PMID- 17402054 TI - Flavonoids from Artemisia scoparia. PMID- 17402056 TI - [The occurrence of hydrocyanic acid in Cormophytes. 18th. Communication. New observations in leguminosae-Papilionoideae (Galegeae, Genisteae, Loteae, Phaseoleae)]. AB - Cyanogenesis in Galegeae, Genisteae, Loteae and Phaseoleae, four tribes of Leguminosae-Papilionoideae, is discussed thoroughly. Oxytropis campestris (Galegeae) is shown to be facultatively cyanogenic (table I); cyanophoric plants contain a zierin-like glucoside. Lotononis crumaniana (Genisteae) is strongly cyanogenic and contains prunasin. Lotaustralin, accompanied by up to 5% Linamarin was demonstrated to be present in three taxa belonging to Dorycnium pentaphyllum sensu lato (table I). Leaves and stems of Phaseolus lunatus contain linamarin and perhaps trace amounts of lot-australin, but no taxiphyllin- or dhurrin-type glucosides. PMID- 17402057 TI - Phytochemistry of the genus Thapsia. AB - In a chemotaxonomic study of the genus Thapsia (Apiaceae), root material of five taxa has been analysed for skin irritating sesquiterpene lactones and the coumarins, 6-methoxy-7-geranyloxycoumarin and scopoletin. Eight tetraesters of 2, 3, 7, 8, 10, 11-hexahydroxy-C (6)-guaianolide and two triesters of 3, 7, 8, 10, 11-pentahydroxy-C (6)-guaianolide were isolated. One of these, trilobolide, has previously been isolated from Laser trilobum Borkh., whereas the others are new. They represent a new group of selective histamine liberators. Regarding the investigated constituents, T. garganica L. was found to differ from T. transtagana Brot. T. villosa L. proved to be chemically heterogeneous and the examined plant samples could be divided into four chemical groups, named I-IV. Hydroxylated C (6)-guaianolides were isolated from T. garganica, T. transtagana, T. maxima Mill., T. villosa I and III. No skin irritating guaianolides were detected in either T. villosa II and IV or in Rouya (Thapsia) polygama (Desf.) Coincy. 6-Methoxy-7-geranyloxycoumarin showed the same distribution as the guaianolides, whereas scopoletin was found in all the plant samples investigated. PMID- 17402058 TI - Quinolizidine alkaloid composition of plants and of photomixotrophic cell suspension cultures of Sarothamnus scoparius and Orobanche rapum-genistae. AB - The alkaloid composition of Sarothamnus scoparius cell cultures, plants and of its root parasite Orobanche rapum-genistae were analyzed by capillary gas-liquid chromatography (GLC) and capillary GLC/mass spectrometry. S. scoparius plants were found to contain the already known isosparteine, sparteine, 17-oxosparteine, lupanine and other quinolizidine alkaloids such as ammodendrine, N methylangustifoline, alpha-isolupanine, 5,6-dehydrolupanine, 4-hydroxylupanine, 17-oxolupanine and five 13-hydroxylupanine esters. Two new alkaloids could be identified, a dehydrosparteine (presumably the 11,12-dehydroderivative) and a dihydroxylupanine (presumably 4,13-dihydroxylupanine). The distribution of these alkaloids within the different plant organs is reported. Orobanche rapum-genistae plants contain the already known sparteine, lupanine and 13-hydroxylupanine and 12 other lupin alkaloids, which are present in the Sarothamnus host plant. Photomixotrophic cell suspension cultures of S. scoparius accumulate lupanine as the main alkaloid. On tissue differentiation (bud formation) the cultures accumulate sparteine as well as lupanine. Whereas sparteine is stored in the cells, lupanine is excreted into the medium. Alkaloid formation of the cell cultures is positively correlated with chlorophyll content of the cells. The alkaloid pattern found in S. scoparius supports the hypothesis that lupanine biosynthesis is the common route of quinolizidine alkaloid biosynthesis from which the other alkaloids such as sparteine are derived. PMID- 17402060 TI - [New quinones in Rubia cordifolia L. Roots, III]. AB - From roots of Rubia cordifolia L. four anthraquinones are isolated: 1-hydroxy 2 methoxy anthraquinone; 1,4-dihydroxy 2-methyl 5-methoxy anthraquinone or 1,4 dihydroxy 2-methyl 8-methoxy anthraquinone; 1,3-dimethoxy 2-carboxy anthraquinone and rubiadin. PMID- 17402061 TI - [Isolation and properties of Sarothamnoside, a novel isoflavone glycoside from various Sarothamnus species]. AB - Sarothamnoside (genistein 7,4'-di-O-[4-O-beta-D-glucopyranosyl-beta-D apiofuranoside]), a novel isoflavone glycoside, has been isolated from Sarothamnus scoparius and S. patens seeds. The structure, in agreement with acid and enzymatic hydrolysis experiments, has been unambiguously determined, using spectral methods: SM, (1)H NMR and (13)C NMR. (1)H NMR spectra showed the beta configuration of the two apioses, whereas (13)C NMR provided evidence of the sugar linkages. PMID- 17402062 TI - (2E,4E)-N-isobutyl-9-piperonyl-nona-2,4-dienoic amide from Ottonia anisum. AB - Roots and stems of Ottonia anisum contain besides 1-butyl-3,4 methylenedioxybenzene and piperovatine, the novel (2E,4E)-N-isobutyl-9-piperonyl nona-2,4-dienoic amide. PMID- 17402063 TI - Kaerophyllin, a new lignan from Chaerophyllum maculatum. AB - From the methanolic extract of roots of spotted cow parsley (Chaerophyllum maculatum Willd.) a new lignan, named kaerophyllin, has been isolated and identified as A-(trans-3,4-dimethoxybenzylidene)- beta-(3,4 methylenedioxylbenzyl)-gamma-butyrolactone. Its structure has been established on the basis of the analysis of UV, IR-, (1)H-NMR, (13)C-NMR and mass spectra. PMID- 17402064 TI - Anthraquinone pigments from Cassia javanica seeds. AB - From the seeds of Cassia javanica chrysophanol, physcion, 1,5-dihydroxy 4,7 dimethoxy 2-methyl anthraquinone 3-O-alpha-L(-) rhamnopyranoside and 1,3,6,7,8 pentahydroxy-4-methoxy 2-methyl anthraquinone have been isolated. Chrysophanol and physcion are commonly occurring anthraquinones but 1,5-dihydroxy 4,7 dimethoxy 2-methyl anthraquinone 3-O-alpha-L(-) rhamnopyranoside and 1,3,6,7,8 pentahydroxy 4-methoxy 2-methyl anthraquinone have not been reported earlier; and in the present communication the structural studies of these are reported. The structures of the two compounds have been arrived at by their colour reactions, spectral data and degradation studies. PMID- 17402065 TI - Alkaloid constituents of Sida acuta, S. humilis, S. rhombifolia and S. spinosa. AB - Three types of alkaloidal constituents, viz., beta-phenethylamines, quinazolines and carboxylated tryptamines, in addition to choline and betaine have been isolated from Sida acuta Burm., S. humilis Willd., S. rhombifolia L., and S. spinosa L. and characterized by their physical and spectral properties, and by chemical transformations. The qualitative and quantitative variations in the alkaloidal constituents of roots and aerial portions at different stages of growth were also noted. Elaboration of the quinazoline alkaloids seems to be a characteristic feature of this genus. The favourable combination of sympathomimetic amines and vasicinone in these species would account for their major therapeutic uses in the Indian system of medicine. PMID- 17402067 TI - The composition of a mixture of ingol esters from Euphorbia kamerunica. PMID- 17402071 TI - Biotransformation of secondary glycosides in Digitalis lanata leaf discs. PMID- 17402070 TI - Phytochemical investigation of Hippeastrum vittatum growing in Egypt. PMID- 17402072 TI - HPLC determination of coumarins in Matricaria chamomilla. PMID- 17402073 TI - Steroidal constituents of Costus afar. PMID- 17402075 TI - Quinolizidine alkaloids from plants and their cell suspension cultures. AB - The alkaloid composition of cell suspension cultures and differentiated plants of Lupinus polyphyllus was evaluated using quartz capillary gas-liquid chromatography, GLC-MS and FD-MS. Lupanine (97% of total alkaloids), sparteine, 13-angeloyloxylupanine and 13-tigloyloxylupanine were detected in alkaloid extracts of L. polyphyllus cell suspension cultures. Lupanine, 13-cis and 13 trans-cinnamoyloxylupanine were found in the spent cell culture medium. No significant difference was found in the alkaloid composition of photomixotrophic and heterotrophic cell strains although the alkaloid content was 5 to 10 times higher in photomixotrophic cell strains. In the respective plants we could identify 18 alkaloids which include the following esters of 13-hydroxylupanine: 13-tigloyloxylupanine, 13-angeloyloxylupanine, 13-cis-cinnamoyloxylupanine, 13 trans-cinnamoyloxylupanine, 13-benzoyloxylupanine, 13-(2-methylbutyryl) oxylupanine; and 13-vanilloyloxylupanine. PMID- 17402076 TI - Further alkaloids from Strychnos dolichothyrsa. AB - The isolation of one of the major alkaloids and 11 minor alkaloids from the stem bark of Strychnos dolichothyrsa is described. The major alkaloid was identified as bisnor-C-alkaloid H. Two new dimeric alkaloids derived from this alkaloid were found, dolichocurine and dolichothine. For the former alkaloid a structure is proposed. Furthermore, the following minor alkaloids were identified: bisnor-C alkaloid H N-oxide and di-N-oxide, Wieland Gumlich aldehyde, 18-deoxy Wieland Gumlich aldehyde, 11-methoxy-diaboline, tubotaiwine, condylocarpine, nor-C fluorocurarine and nor-macusine B. In the extracts of the stem bark of S. urceolata 11-methoxydiaboline was identified as one of the major alkaloids. PMID- 17402077 TI - Gynocardin and tetraphyllin B from Carpotroche brasiliensis. AB - Gynocardin and Tetraphyllin B were isolated from the seeds and pericarp of Carpotroche brasiliensis (Raddi) Endl. (Flacourtiaceae). The cyanogenic glycosides occur in an approximately equimolar mixture in the seeds and in the pericarp. Their structures were confirmed by (1)H-NMR. PMID- 17402078 TI - Isolation of the secoiridoid-glucoside sweroside from Tabernaemontana psorocarpa. AB - The known secoiridoid glucoside sweroside (structure I) has been isolated from the leaves of Tabernaemontana psorocarpa and identified by chemical, chromatographic and spectroscopical data. Its (1)H-NMR at 360 MHz is discussed. PMID- 17402079 TI - Molluscicidal properties of various saponins. AB - A series of 24 different saponins isolated from various medicinal plants have been tested against Biomphalaria glabrata, one of the snail vectors of schistosomiasis (bilharziasis). In general, monodesmosidic triterpenoid saponins exhibit strong molluscicidal activity whereas bidesmosidic saponins as well as the aglycones are fully inactive. Some dammarane glycosides and alkaloidal saponins have also been tested. PMID- 17402080 TI - [New cardioactive drugs II, detection and isolation of cardiotonic amines with ionpair-HPLC]. AB - An ionpair-HPLC-technic using a reversed-phase column was developed to screen plant extracts for cardiotonic amines and to isolate them in a preparative scale for direct MS and pharmacological investigations. By this method the positive inotropic tyramine could be isolated from Selenicereus grandiflorus (L.) Britt et Rose (Cactus grandiflorus). PMID- 17402081 TI - Influence of strain specificity and culture conditions on terpene production by fungi. AB - The volatile oil production of several species and strains of the genus Ceratocystis (Ascomycetes) and of Lentinus lepideus (FR.) FR. (Basidiomycetes) is studied. The production of mono- and sesquiterpenes by these fungi proves to be highly influenced by strain specificity and culture conditions. PMID- 17402082 TI - [A chemical study of the essential oil from Artemisia arborescens]. AB - From Artemisia arborescens L. (Compositae-Anthemidae) essential oil, (+)-beta thuyone (main compound), chamazulene and caryophyllene-epoxyde have been isolated; 14 other mono-and sesquiterpenes have been identified. Chemotaxonomical interest of some of the essential oil constituents is discussed. PMID- 17402083 TI - [New flavonoid-glycosides from Crataegus monogyna and Crataegus pentagyna]. AB - From the leaves and flowers of Crataegus monogyna and C. Pentagyna six new flavonoid-C- and O-glycosides respectively have been isolated and identified as 2''-O-rhamnosyl-orientin, 2''-O-rhamnosyl-isoorientin, 2''-O-rhamnosyl isovitexin, rutin, spiraeosid, 8-methoxy-kampferol and 8-methoxy-kampferol-3-O glucoside. The structure of O-rhamnosyl-vitexin and O-acetyl-O-rhamnosyl-vitexin isolated previously, have been elucidated unambigiously mainly by NMR- and MS spectroscopy. PMID- 17402084 TI - Radioimmunoassay for the citrus bitter principle, naringin, and related flavonoid 7-O-neohesperidosides. AB - An immunoassay for the citrus bitter principle, naringin, and related flavonoid-7 O-neohesperidosides is reported. The assay detects ca. 2 ng of naringin and can be used to quantify this compound in the parts per billion (ppb) range in crude grapefruit juice and extracts of other plant tissues. The antiserum used is highly reactive with the 2-rhamnosyl-1-glucopyranose at the C-7 position but not with e.g. the isomeric 6-rhamnosyl-1-glucopyranose moiety and can, thus, be used to identify the stereochemistry of this disaccharide moiety at the C-7 position of flavanoids. The assay involves a directly iodinated naringin-[ (125)I] as immunotracer. PMID- 17402085 TI - [Constituents of the essential oil of myrrh]. AB - From the essential oil of myrrh, Commiphora abyssimica (Berg) Engler (Burseraceae) have been isolated: nine sesquiterpenoid hydrocarbons, the sesquiterpene alcohol elemol and the furanosesquiterpenoids furanodiene, furanodienone, isofuranogermacrene, curzerenone and lindestrene. PMID- 17402086 TI - Alkaloids of Vinca major cv. Variegata. AB - Analysis of the leaves of Vinca major L. cv. Variegata led to the isolation of seven alkaloids: reserpinine, majdine, akuammicine, strictosidine lactam, pseudoakuammigine, akuammine, and a new alkaloid tentatively assigned as 10 hydroxycathofoline. The yellowish and green laminar sections of the leaves did not differ in their alkaloid profile. PMID- 17402087 TI - Determination of flavonoids in seeds of Zizyphus vulgaris var. spinosus by high performance liquid chromatography. AB - A new method for separation and quantitative determination of flavone-C glycosides in seeds of Zizyphus vulgaris var. spinosus by high performance liquid chromatography was elaborated. A reverse-phase system with a COPELL-ODS column using tetrahydrofuran in water (4:53) as a mobile phase was developed. The flavonoids were detected at 330 nm and the analysis was successfully carried out within 15 min. PMID- 17402088 TI - 6-O-beta-glucopyranosylaucubin, a new iridoid from Odontites verna. AB - Fourteen iridoid glycosides have been isolated from Odontites verna ssp. serotina (Scrophulariaceae). In this paper the isolation of 6-O-beta-glucopyranosylaucubin is described; its structure and configuration has been demonstrated mainly by the aid of the (1)H- and (13)-C-NMR spectroscopy. The existence of a gamma glycosylation shift is showed. PMID- 17402089 TI - Growth and diosgenin production by Dioscorea deltoidea cells in batch and continuous cultures. AB - This paper reports the kinetics of growth and diosgenin production in batch cultures, and the application of the continuous culture (chemostat) technique to Dioscorea deltoidea cells. In batch cultures, biomass production was dependent on the concentration (up to 60 g/liter) of the carbon source (sucrose); the cellular yield value obtained was 0.4 g cell dry wt/g sucrose utilized. Diosgenin was synthesized only after growth had ceased; its synthesis proceeded for about 18 days and a concentration of 1.8% (of cell dry wt.) was obtained. D. deltoidea cells were grown at steady state conditions, at a constant growth rate in a chemostat. Only small amounts of diosgenin were produced by growing cells in the chemostat. PMID- 17402090 TI - The effect of medium constituents on growth and diosgenin production by Dioscorea deltoidea cells grown in batch cultures. AB - The effect of nitrogen and carbon sources on growth and diosgenin production was studied with Dioscorea deltoidea cells which were grown under defined conditions in a chemostat, and transferred to batch cultures. Both ammonium and nitrate nitrogen were required for growth and steroid production; diosgenin production was maximal at higher amounts of nitrogen. The nature and concentration of the carbon source (sugar) had marked effects on diosgenin production. A maximal amount of diosgenin (3.8% of dry wt.) was produced on sucrose (15 g/liter) after 21 days of growth in a batch culture. PMID- 17402091 TI - Composition of essential oils of Laurus nobilis, L. nobilis var. angustifolia and Laurus azorica. AB - The essential oils of Laurus nobilis, L. nobilis var. angustifolia and L. azorica were investigated by means of GLC, IR, PMR, CMR and GC-MS. The composition of the oils of the first two mentioned species was about the same with as main constituent 1,8-cineole, whereas the oil of L. azorica was different with low content of 1,8-cineole (10% vice 40-50%), absence of spathulenol, high content of alpha- and beta-pinene and presence of alpha-copaene, cinnamyl acetate, allo aromadendrene, beta-eudesmol, beta-caryophyllene oxide and 1 (12), 8 (15) caryophylladiene-9alpha-ol. About 50 components were tentatively identified. PMID- 17402092 TI - A plausible biogenetic proposal for koumine. PMID- 17402093 TI - Qualitative changes in peptide alkaloid biosynthesis of Claviceps purpurea by environmental conditions. PMID- 17402094 TI - Incorporation of thiazolidine-4-carbozylic acid into ergosine by Claviceps purpurea. PMID- 17402095 TI - 16-anhydro-3-acetyl-gitoxigenin in Adenium abesum. PMID- 17402096 TI - Dammarane saponins of flower buds of Panax notoginseng (Sanchi-Ginseng). PMID- 17402097 TI - Studies on the constituents of Artemisia annua Part II. AB - The present paper is a continuation of our study on the Chinese traditional herb Artemisia annua L. [1-5], describing several additional constituents: quinghaosu IV and V (V, VII), quinghao acid (VIII) [6], chrysosplenol (VIa) [7] and a paraffinic alcohol; V, VII and VIII are compounds with unreported structures. PMID- 17402098 TI - Degradation of ginseng saponins under mild acidic conditions. AB - Ginseng saponins, ginsenosides Rg (1), Re and Rb (1), decomposed under mild acidic conditions to yield prosapogenins. The structures of the prosapogenins were investigated by (13)C-NMR spectroscopy and Rg (1)-prosapogenin II was shown to be a mixture of ginsenoside Rh (1), and its C-20 epimer, produced by hydrolysis followed by epimerization at C-20. Rg (1)-prosapogenin III, the other prosapogenin derived from ginsenoside Rg (1); was a C-25,26 hydrated derivative of Rg (1)-prosapogenin II. Re-prosapogenin II was identified as a mixture of ginsenoside Rg (2) and its C-20 epimer, and Re-prosapogenine III as a C-25,26 hydrated derivative of Re-prosapogenin II. PMID- 17402099 TI - [Iridoids from Verbascum sinuatum.]. AB - From the underareal part of Verbascum sinuatum harpagoside [1] and a new diacyl iridoid-diglycoside [2] has been isolated. The structure of the new compound is elucidated on the basis of (1)H-NMR, (13)C-NMR, mass spectral data and chemical degradation as 6-O-(2'',3''-Di-O-acyl)-alpha-L-rhamnopyranosylcatalpol. PMID- 17402100 TI - Two new arylnaphthalide lignans from Justicia flava roots. AB - Orosunol and 8-demethylorosunol, two new 1-aryl-2,3-naphthalide lignans, have been isolated from Justicia flava root. Structures IV and V respectively have been proposed for them by spectral (UV, IR, PMR, (13)C NMR, and MS) and chemical evidence. PMID- 17402101 TI - [Flavones libres de Artemisia mesatlantica]. PMID- 17402102 TI - Flavonoids from Dittrichia viscosa. AB - The isolation and structural elucidation of two flavonoids I and II found in Dittrichia viscosa, L., syn. Inula viscosa L., is reported. PMID- 17402103 TI - [Furocoumarins in Archangelica officinalis and Angelica silvestris. Distribution of 5-beta-cyclolavandulyloxy-psoralen in the genus Angelica.]. AB - 5-beta-Cyclolavandulyloxy-psoralen i.e. 4-[(2,4,4-trimetnyl-1-cyclohexen-1 yl)methoxy]-7H-furo [3,2-g] [1] benzopyran-7-one was isolated from Angelica silvestris L. Detailed analytical data are given. Presence of this coumarin in other Angelica species is reported and discussed. PMID- 17402104 TI - A new commercial strain of ergot adapted from a wild grass. AB - A strain of Claviceps purpurea (Fr.) Tul. isolated from Arthraxon lancifolius Hochst. was successfully adapted on rye. Experimental cultivation on rye showed 370 kg/ha yield of sclerotia containing 0.5% total alkaloid which consisted of 33% ergometrine, 17.6% ergotamine, 18.7% ergocornine and 22.7% ergokryptine. The results also reveal that alkaloid profile of the ergot strain depends largely on its host. Significantly high yield of the sclerotia and the valuable alkaloid profile of the new strain indicate its commercial potential. PMID- 17402105 TI - Alkaloids of Corydalis slivenensis. AB - The known alkaloids (-)-stylopine, (-)-canadine, (+/-)-sinactine, (+)-nantenine, (+)-bulbocapnine, protopine, (-)-isocorydine, (-)-domesticine, alpha allocryptopine and berberine were found in a mixture of alkaloids from Corydalis slivenensis (Papaveraceae), growing in Bulgaria. Dehydronantenine, corydine, predicentrine and isoboldine were detected by means of a thin-layer chromatography. Two other alkaloids - (+)-tetrahydrocorysamine and (-)-cavidine, were identified on the basis of their spectral data. PMID- 17402106 TI - Anacardic acid: molluscicide in cashew nut shell liquid. AB - The components of anacardic acid, (a mixture of 6-n-C (15) alkylsalicylic acids whose side chains vary in degrees of unsaturation) have been isolated by high pressure liquid chromatography from a crude extract of cashew nut shell, Anacardium occidentale, and tested for toxicity to fresh water snails, Biomphalaria glabrata. The triene component is the most toxic form (LC (50) 0.35 ppm), the diene and monoene components are less toxic (LC (50) 0.9 and 1.4 ppm), and the saturated component is relatively nontoxic (LC (50) > 5 ppm). Since decarboxylated anacardic acid (cardanol) and salicylic acid do not kill snails at concentrations up to 5 ppm, it appears that both, carboxyl group and unsaturated side chain are absolutely required for molluscicidal activity. The mechanism of toxicity of anacardic acid to snails is unknown. PMID- 17402107 TI - [Zierinxyloside, a new Cyanogenic glycoside from Xeranthemum cylindraceum.]. PMID- 17402108 TI - Boerhaavia diffusa, a new source of phytoecdysones. PMID- 17402109 TI - Genus crotalaria. PMID- 17402110 TI - [Pyrrolizidinalkaloide aus Senecio congestus]. PMID- 17402111 TI - Neosenkirkine and Senkirkine from Senecio pierotii. PMID- 17402112 TI - Alkaloids of the Genus Datura, Section Brugmansia. PMID- 17402113 TI - Chemical Constituents of Centaurea brugueriana. PMID- 17402114 TI - 4,5-Dihydro-11-NOR-11-Hydroxy-Delta 7,11-Santonin, A New NOR-Sesquiterpene-gamma Lactone Found in Crepis pygmaea. PMID- 17402115 TI - [The ocurrence of Viridiflorol in Salvia officinalis.]. PMID- 17402116 TI - Dihydropiplartine from Piper bartlingianum. PMID- 17402117 TI - [DC- and HPLC-Analysis of Eleutherococcus.]. AB - A rapid and reproducible TLC- and HPLC-analysis of root-extracts and preparations of ELEUTHEROCOCCUS SENTICOSUS on the basis of the characteristic lignans and other phenylpropan derivatives has been developed. PMID- 17402118 TI - Studies on coumarins of a chinese drug "Qian-Hu". AB - Three new and four known coumarins were isolated from a Chinese drug "Zi-Hua Quian-Hu" (Shikazenko in Japanese), the root of Peucedanum decursivum Maxim. (Umbelliferae). The new compounds were proved to be 3'(S)-senecioyloxy-4'(R) hydroxy-3',4'-dihydroxanthyletin, 3' (S)-hydroxy-4' (R)-senecioyloxy-3',4' dihydroxanthyletin and 3'(S)-angeloyloxy-4'(R)-acetoxy-3',4'-dihydroxanthyletin. The known coumarins were identified to be decursidin, bergapten, nodakenetin and nodakenin, respectively. PMID- 17402119 TI - Two new iridoid glucosides of Plantago asiatica. AB - From the whole plant of Plantago asiatica, two new iridoid glucosides were isolated and their structures were established as 3,4-dihydroaucubin and 6'-O beta-glucosylaucubin, respectively. PMID- 17402120 TI - Phthalides in the essential oil from roots of Levisticum officinale. AB - The composition of the phthalide mixture of the essential oil from roots of Levisticum officinale Koch was investigated. The phthalide mixture was analyzed by combining separation methods - GLC, LSC and TLC - and subsequently using spectroscopic methods - IR, MS and NMR. E- and Z-butylidenephthalide, E- and Z ligustilide, senkyunolide and validene-4,5-dihydrophthalide were found to be present; isosenkyunolide and propylidenephthalide were tentatively identified. The influence of the isolation procedure on the composition of the phthalide mixture was tentatively studied by comparison of mixtures isolated by solvent extraction and hydrodistillation respectively. PMID- 17402121 TI - Monomeric Indole Alkaloids from Ervatamia hainanensis. AB - Eleven monomeric indole alkaloids have been isolated from the roots of Ervatamia hainanensis. Nine of them are known: coronaridine 1, coronaridine hydroxyindolenine 2, heyneanine 3, vobasine 4, perivine 5, ibogamine 6, geissoschizol 7, 10-hydroxygeissoschizol 8 and 3-oxocoronaridine 9; and two are new: 10-hydroxyheyneanine 10 and 3-(beta-hydroxyethyl)-coronaridine 11. PMID- 17402122 TI - [Oleoresin of Aucoumea klaineana.]. AB - From oleoresin of Aucoumea klaineana Pierre, five triterpenes are isolated; one pentacyclic triterpene: alpha-amyrin and four tetracyclic triterpenes: masticadienediol which seems to be new in this family, 3alpha-hydroxytirucalla 8,24-dien-21-oic acid, 3 alpha-hydroxytirucalla-7,24-dien-21-oic acid which have been already isolated in some Burseraceae and 3-oxotirucalla-7,24-dien-21-oic acid which seems to be a new triterpene. PMID- 17402123 TI - [Investigations on harpagophytum.]. AB - Comparing the analytical results of the constituants of naturally growing roots and callus of Harpagophytum procumbens, it could be demonstrated that both, the products of the primary and the secondary metabolism, showed important differences. Harpagosid which is present in significant amounts in the roots and tubers of the fresh plants, was shown to be completely absent in the callus. Stachyose the main reserve carbohydrate again was only produced in minor amounts. Fructose was the predominant sugar in the callus cells. PMID- 17402124 TI - Virgidivarine, a New Alkaloid of Virgilia divaricata. AB - Virgidivarine, C (15)H (26)N (2)O (2), is a new dipiperidine alkaloid, isolated from the South African Virgilia divaricata Leguminosae. Its systematic name is 3 carboxy-N-(4'-butenyl)-5-(2'-piperidyl) piperidine. PMID- 17402125 TI - Virgiboidine, A New Alkaloid from Virgilia oroboides and Virgilia divaricata. AB - From the leaves of the South African Leguminosae Virgilia oroboides and V. divaricata a new alkaloid virgiboidine, C (15)H (24)N (2)O, has been isolated by means of column chromatography. In these species 15 more alkaloids are detected, of which 6 are identified including lupinine. PMID- 17402126 TI - Partial purification and properties of tropine dehydrogenase from root cultures of Datura stramonium. AB - From sterile root cultures of Datura stramonium, an NADP(H)-specific tropine dehydrogenase has been isolated and characterized. The enzyme catalyzes the reversible and stereospecific oxidation of tropine and related tropane-3 alpha ols to the corresponding ketone. Isomeric pseudotropine (tropane-3 beta-ol) is neither accepted as substrate nor produced in the reverse reaction. It is assumed that this dehydrogenase is involved in the biosynthesis of tropane alkaloids. PMID- 17402127 TI - Efficient incorporation of 10-hydroxygeraniol and 10-hydroxynerol into indole alkaloids by a cell suspension culture of Catharanthus roseus. AB - A cell suspension culture of Catharanthus roseus was evaluated with regard to its potential usage in studies of early stage indole alkaloid biosynthesis. Employing this material, 1,1,7-trideutero-10-hydroxygeraniol and 1,1,7-trideutero-10 hydroxynerol were incorporated into the alkaloids ajmalicine and strictosidine lactam in approximate proportions of 50% and 80% respectively. PMID- 17402128 TI - Tissue culture of Cinchona pubescens: effects of media modifications on the growth. AB - Studies were performed on the growth of callus cultures of CINCHONA PUBESCENS V AHL concerning the influence of the concentration of major and minor elements described by M URASHIGE and S KOOG [1], as well as a number of organic constituents in the medium. After 13 weeks a maximum increase of the fresh and dry weight was observed in tissues grown on a medium containing 2,4-D (1 mg/l) and BA (2 mg/l) as growth factors and with the same or the half concentration of mineral salts as used by M URASHIGE and S KOOG. PMID- 17402130 TI - [Flavonoide in den Blattern von Nelia meyeri]. PMID- 17402129 TI - Constituents of Magnolia grandiflora, cyclocolorenone. PMID- 17402133 TI - [Distal radius fracture]. PMID- 17402134 TI - [Distal radius fractures: new concepts as basis for surgical treatment]. AB - New advances in the biomechanics and pathomechanics of distal radius fractures as well as new generations of plates and improved surgical approaches now make possible the stable management and early functional rehabilitation not only of simple but also of complicated distal radius fractures according to the principles for articular and juxta-articular fractures. Especially for complex articular fractures, the fracture patterns are so heterogeneous that an individual surgical treatment strategy must be developed for each case. The preoperative clarification of articular fractures with computed tomography is particularly useful for planning surgery. Mental exposition with the three-column model and pathomechanics is an important prerequisite for understanding this type of injury and the appropriate choice of operative technique. A decisive factor for success is the subtle surgical procedure in approaching and handling the fine plates. PMID- 17402135 TI - [Mal-united fractures of the distal radius--biomechanics and operative treatment options]. AB - Despite all progress in the treatment of distal radius fractures, malunion is still one of the most common complications. It may be true that not all non anatomically aligned fractures of the distal radius are causing disabilities. However, most patients with a distal radial malunion complain about loss of wrist motion and forearm rotation, loss of grip strength, and pain. These complaints should lead to a sophisticated clinical examination and a careful evaluation of the radiographs of the wrist, and - if needed - to further investigations. Treatment options are procedures solely aimed to diminish pain, procedures aimed to improve wrist and forearm function without restoration of the anatomic relationships, and procedures aimed to restore the anatomy of the wrist joint, especially corrective osteotomy of the radius. Radial corrective osteotomy improves significantly wrist and forearm function, grip strength, and diminishes pain. The clinical results are as good or better than the radiological results. PMID- 17402136 TI - [Distal radius fractures--aetiology, treatment and outcome]. AB - INTRODUCTION: Distal radius fractures were investigated in a retrospective open multicenter cohort study to assess aetiology, fracture pattern and treatment modalities and their influence on subjective and objective outcome. PATIENTS AND METHOD: Demographic data, fracture history, course of fracture healing, functional and radiological parameters and the DASH-questionnaire were collected from 18 Austrian hospitals and analysed statistically. RESULTS: PATIENTS: n = 707, 465 (65.8 %) female, 242 (34.2 %) male. Mean age: 52 (19 - 86) years; age group 1 (19 - 39 years) 26 %, age group 2 (40 - 59 years) 41 %, age group 3 (60 years and elder) 33 %. FOLLOW-UP: mean 5.8 (3.9 - 17) years; HISTORY: fall 65.1 %, sports 17.4 %, traffic accidents 8.9 %, fall from great height 7.5 %; others 1.1 %. Most frequent fracture patterns according to AO: A2 (26.6 %), C2 (22.2 %), A3 (16.1 %), C1 (12.7 %); according to PE: I-2 (44.8 %), I-1 (40.0 %), III-2 (4.4 %), II-2B (4 %). There was no significant correlation between fracture pattern and age groups for both fracture classifications. TREATMENT: 57.9 % surgical, 42.1 % conservative. Radiological results: Depending on treatment, there were significant differences between the radial tilt and the palmar radial inclination, the dorsal and palmar ulnar variance and the width of the DRU-joint. DASH-questionnaire: Median 6.03 (90 - 0). There was a significant, minor positive correlation of bad results in the elderly and a minor positive correlation of bad results correlated to the palmar radial inclination as well as a minor negative correlation to the palmar DRUJ-value and the ulnar variance. CONCLUSION: The incidence of distal radius fractures was increased in females and in patients with the age between 40 - 59 years. The most frequent cause to sustain a distal radius fracture was a simple fall. 85 % of the fractures were dislocated dorsally. The most frequent fracture type was the dorsal intraarticular. There was no specific fracture type observed to be typical for one of the age groups and surgical treatment was almost as frequent as conservative. Surgical treatment improved reconstruction of the radial tilt, palmar inclination and the DRU-joint. Subjective outcome was worse in elderly patients. The radiological changes in the DRU-joint correlated to minor DASH values. PMID- 17402137 TI - [Multidirectional palmar fixed-angle plate fixation for unstable distal radius fracture]. AB - PURPOSE/BACKGROUND: Open reduction and internal fixation with plates is a valid treatment of displaced extra- and intra-articular distal radius fractures. This report presents our experience treating unstable distal radius fractures by using a multidirectional palmar fixed-angle plate system (Aptus(R) Radius 2.5 by Medartis). The subchondral support of the articular surface by fixed angle screws prevents secondary dislocation allowing early mobilisation. PATIENTS AND METHOD: 61 patients with distal radius fractures were treated during 6/2003 and 10/2004 using this palmar fixed-angle plate fixation without bonegraft. Postoperative X rays showed anatomical reduction in all cases. 55 patients could be evaluated postoperatively. The follow-up examination included the patient's history, physical and radiographic examination as well as the DASH questionnaire and the Krimmer's modified Cooney wrist score. RESULTS: The average follow-up time was 9.6 +/- 5.2 months. The mean age of the examined patients was 53.9 +/- 17.8 years. The fractures were classified according to AO. There were eleven A3, four B2 and five B3 fractures, ten C1, 16 C2 and nine C3 fractures. All fractures united without complications. Just two cases showed a loss of length. At follow up the average palmar angulation was 8 degrees , ulnar inclination 21 degrees and ulnar variance + 0.2 mm. Wrist motion averaged a decrease for extension and flexion of 12 %, for ulnar and radial deviation of 9 % and for pronation and supination only of 2 % in comparison to the uninjured side. Grip strength reached an average of 85 % of the contralateral side. Patients regained good function as represented in a mean DASH score of 14 points and a Krimmer score of 82 points. CONCLUSIONS: The treatment of unstable distal radius fractures with this plate fixation provided stable internal fixation and allowed early function. Due to multidirectional angle fixation and two lines of cortical screws it is possible to achieve an optimal restoration. The fixation of the central articular surface is guaranteed by the distal line, the dorsal subchondral support by the proximal line. PMID- 17402138 TI - [Limits of palmar locking-plate osteosynthesis of unstable distal radius fractures]. AB - PURPOSE: The use of locking plate systems in the treatment of distal radius fractures has increased during the last years. In the presented study our experience and results after palmar plating of dorsal dislocated distal radius fractures are analysed and presented. Besides the functional and radiological results, the potential surgical risks and the observed complications are discussed. PATIENTS AND METHODS: Over a mean 15-month period, 112 consecutive patients (24 men, 88 women) with an average age of 57 years who were treated for an unstable dorsal dislocated distal radius fracture using the palmar locking plate system could be assessed. The functional results were compared with the uninjured contralateral side. The subjective pain level was assessed using the visual analogue scale (VAS) and the subjective results were assessed using the Disabilities of the Arm, Shoulder, and Hand (DASH) Score. Radiographic assessment included palmar tilt, radial inclination, and ulnar variance. The objective and subjective functional results were evaluated using the modified Mayo-Wrist Score. RESULTS: Functional results: extension slashed circle 53.1 degrees (84 % as compared with the uninjured side); flexion slashed circle 45.8 degrees (81 % as compared with the uninjured side); pronation slashed circle 78.7 degrees (95 % as compared with the uninjured side); supination 81.9 degrees (95 % as compared with the uninjured side); grip strength slashed circle 56 Kp (73 % as compared with the uninjured side). Radiological results: postoperative reduction/mean postoperative loss of reduction: radial inclination: slashed circle 20.4 degrees /slashed circle 0.2 degrees ; palmar tilt: slashed circle 0.2 degrees /slashed circle 3.1 degrees ; ulnar variance: 0.08 mm/slashed circle 0.42 mm. Pain values: slashed circle 2.7 points; DASH score: slashed circle 14.8 points; complications: breakage of plates 1 %, screw loosening 3 %, intraarticular screw position 2 %, delayed bone union 4 %, rupture M. extensor pollicis longus 2 %/M. flexor pollicis longus 3 %, peritendinitis extensor tendons 5 %, synovialitis flexor tendons 8 %, CTS 3 %, complex regional pain syndrome 6 %. CONCLUSION: Favourable indications for palmar locking plate osteosynthesis are A2, A3, C1 and C2 fractures with big distal fracture fragments. In these cases, additional bone grafting is not needed. In very distal fractures, multidirectional plate-systems are advantageous. Multifragmental C3 fractures show a high complication rate. Additional bone grafting for the metaphyseal defect should be considered. PMID- 17402139 TI - [Is corrective osteotomy for malunited distal radius fractures also indicated for elderly patients?]. AB - BACKGROUND: Corrective osteotomy of the distal radius is a well accepted procedure in younger, manual working and active patients with a malunited distal radius fracture. These functional improving procedures are often abandoned in elderly patients on accout of an uncertain benefit. PURPOSE: To investigate the radiological and clinical outcome after corrective osteotomy for malunited fractures of the distal radius in the elderly and to answer the question, whether there is a benefit or not. METHODS: In a prospective study the results of corrective osteotomy for malunion of distal radius fractures in 17 patients aged 65 years and more were investigated and compared with the results of 31 patients below an age of 65 years. Radiological parameters were radius tilt, ulnar inclination and ulnar variance. Clinically wrist motion, forearm rotation, pain at activity and grip strength were measured. Both groups were comparable according to their preoperative data. The statistical analyses were performed with use of the Mann-Whitney-U-test and the Wilcoxon test for paired differences, accepting p < 0.05 as significant. RESULTS: All osteotomies achieved bony union. Loss of correction or other complications did not occur. Postoperatively both groups demonstrated improvement of all clinical and radiological parameters. Improvement was significant in eleven of twelve parameters in the group below 65 years and for eight of twelve parameters in the elderly. A comparison among the two groups revealed significant better results for five of the twelve parameters in the younger patients. CONCLUSION: Radial corrective osteotomy leads to an improvement of the clinical situation not only in the younger but also in the elderly patient. Therefore, radial corrective osteotomy can be recommended for elderly patients. However, elderly patients may not experience the same amount of benefit from a corrective osteotomy of the distal radius as the younger patients. An age-related higher complication rate or unsatisfying long term results were not detected. PMID- 17402140 TI - [Sagittal wrist motion following dorsal radiocarpal fracture dislocations]. AB - PURPOSE: Radiocarpal fracture dislocation is a rare, complex injury characterised by dislocation of the radiocarpal joint with avulsion of the dorsal or palmar cortical margin of the distal radius. To evaluate the sagittal motion at the radiocarpal and midcarpal levels following dorsal radiocarpal fracture dislocation (Moneim type I) nine cases were investigated clinically and radiologically. PATIENTS AND METHOD: In a retrospective follow-up examination, eight patients could be included. The average follow-up was 4.1 years. One patient had a bilateral injury. The operative approach was bilateral in all cases. Restoration of the radial articular surface, filling metaphyseal defect zones with cancellous bone graft and internal fixation with a special T-plate were performed from dorsal. For refixation of the radiocarpal ligaments a small palmar approach was used. Standard anteroposterior and lateral radiographs, as well as lateral views in full extension and flexion were taken at follow-up. Clinical investigation included measurement of active range of motion, grip strength and pain evaluation using the VAS. RESULTS: Radiological evaluation of the standard lateral view turned out a mean angle between scaphoid and lunatum of 55.6 degrees, capitatum and lunatum of -11.6 degrees and radius and capitatum of 10.5 degrees. In full flexion the following angles were measured: radius/lunatum 15.3 degrees, capitatum/lunatum 18 degrees and between radius and capitatum 30 degrees. In full extension the angles averaged: radius/lunatum -23.9 degrees, capitatum/lunatum -31 degrees and between radius and capitatum -55 degrees. According to the Knirk and Jupiter classification system, five patients presented arthritis stage 1, three arthritis stage 2 and one a stage 3 arthritis. Clinical evaluation showed a mean wrist motion of 55 degrees for extension, 35 degrees for flexion, 88 degrees for pronation, 70 degrees for supination and 25.5 degrees for the mean radial as well as the mean ulnar motion. The average Mayo Wrist Score was 76.1 points. CONCLUSION: Operative treatment of dorsal radiocarpal fracture dislocation using a bilateral approach led to satisfying results in eight of nine cases with decreased but radiologically evaluated sagittal motion of the proximal row. PMID- 17402141 TI - [Long-term results following ORIF of dorsal dislocated distal intraarticular radius fractures]. AB - PURPOSE: To evaluate the sequelae of distal intraarticular radius fracture with regard to the development of arthritis and clinical symptoms. PATIENTS AND METHOD: In a retrospective follow-up examination, 72 patients with a distal intraarticular radius fracture could be included for clinical and radiological investigation 9 years following the trauma. All fractures were treated by ORIF and cortico-cancellous bone grafting. RESULTS: Radiological evaluation revealed 5.1 degrees palmar tilt, 19.1 degrees radial tilt and the ulnar variance amounted to -0.5 mm. The articular cavity depth in the sagittal plane measured with 4.6 mm, 1.2 mm more than the non-involved side. Articular step-off was noticed in 6 patients. According to the Knirk and Jupiter classification system, two patients healed without arthritis, 43 patients presented arthritis stage 1, and 27 stage 2. Evaluation of the data showed a significant correlation between arthritis and articular cavity depth. But arthritis had neither influence on the DASH, nor the pain level. On the other hand, arthritis led to decreased sagittal wrist motion. CONCLUSION: ORIF of distal intraarticular radius fractures led to predictable results concerning restoration of length and form of the distal radius. Arthritis had a minor influence on the clinical end result. PMID- 17402142 TI - [MR imaging for the evaluation of accompanying injuries in cases of distal forearm fractures in children and adolescents]. AB - INTRODUCTION: The study was made to evaluate the role of MR imaging in pediatric distal forearm fractures by comparison with the findings of plain radiographs and MRI. MATERIAL AND METHOD: 38 patients (27 boys and 11 girls, mean age of 12 years, range 7 to 15 years) with radiographically open distal radius and ulna growth plates requiring first aid for a fracture of the distal third of the forearm, were included in this study. The fractures were diagnosed on plain radiographs and conservative treatment was performed. In 35 patients MR imaging was performed within 3 weeks after the accident and in 3 patients MRI was performed after 6 to 9 weeks because of persistent wrist pain. RESULTS: Fifteen Salter/Harris II injuries of the radius and 1 of the ulna, 1 torus fracture of the radius and 2 of the ulna, 12 greenstick fractures of the radius and 3 of the ulna, 10 complete displaced radius fractures and 15 ulnar styloid fractures were found on plain radiographs. Twelve patients had evidence of associated triangular fibrocartilage complex (TFCC) lesions in MRI, there was no statistical correlation between TFCC lesions and fracture types, fracture dislocations or patients age (p > 0.5). One patient had an avulsion of the radioscaphocapitate ligament from the radius accompanying a greenstick fracture of the distal radius. 19 bone bruises and two radiographically occult fractures were identified. In 2 patients, a bone marrow oedema was seen in the radial epiphysis immediately adjacent to the germinal zone of the growth plate. In these patients premature physeal arrest occurred. CONCLUSION: MRI plays an important role in the evaluation of acute pediatric wrist injuries. It allows a better evaluation of osseous lesions than plain radiographs. In our study, a tear of the triangular fibrocartilage complex accompanied distal radius fractures in 32 % of patients. Simultaneous rupture of the TFCC insertion in the fovea ulnaris and ulnar styloid fracture lead to destabilisation of the distal radioulnar joint (DRUJ). PMID- 17402143 TI - [Acceptance of patient-related evaluation of wrist function following distal radius fracture (DRF)]. AB - INTRODUCTION: Outcome evaluation after distal radius fractures is mainly based on wrist function and radiological parameters, while measuring patient satisfaction seems to be more difficult. The purpose of this study was to assess different subjective features with regard to patient satisfaction for wrist healing, activity and participation. MATERIALS AND METHODS: A questionnaire combining items from the established PRWE (Patient related wrist evaluation), wrist specific items from the DASH (Disability of Arm, Shoulder and Hand) as well as the ICF questionnaire (international classification of function, health and disease) was created, and then distributed to patients without wrist injuries (group 1) and patients following a distal radius fracture (group 2). Analysis of answered questions was performed with regard to response rate, valid content, mean differences between the two groups, correlation with radiology features in the patient group and age. RESULTS: Of 510 individuals with non-relevant wrist injuries, 96.7 % of the questions were answered, while of 133 distal radius fracture patients, 92.2 % of the questions were answered, rendering a total response rate of 95.5 %. Frequency of not responding to questions varied between 1.4 % to 14.4 %, and remained < 5 % for 17 questions. Internal consistency of the questionnaire with regard to wrist function was high in both groups (Cronbach alpha index for patients with non-relevant wrist injuries 0.9836, for distal radius fracture patients 0.9881). All questions were deemed specific for wrist function (discriminatory power > 0.7), and highly significant (p < 0.01) for ascertainment of subjective comfort after distal radius fracture in comparison with non-relevant wrist injuries. Fourteen of 25 questions (56 %) were correlated significantly (p < 0.05) with radiological features of malunion. For ages 30 to 49, and > 70 years, wrist function was deemed worse in the distal radius fracture patients vs. the non-injured wrist group. For ages 50 to 69, a significant difference was found only for 5 of 25 (20 %) questions. CONCLUSION: All questions were of a high validity and significance to determine subjective outcome after distal radius fractures. Patient acceptance was significantly different for questions, and subjective wrist comfort depended on age. For ages 50 to 69, additional questions in terms of employment situation and recreational function could be incorporated. Only about half of the questions correlated with radiological parameters of distal radius malunion. Eight questions showed both high acceptance and high correlation with radiology. Standardised measurements of range of motion and power, as well as radiology features and subjective questions of high acceptance and radiology relevance are essential for a wrist-specific questionnaire. PMID- 17402144 TI - [Stabilisation of distal radius fractures by a novel endomedullary, fixed-angle plate: first experience]. AB - BACKGROUND: Based on the fact that not every extra-articular distal radius fracture is stable, we asked ourselves whether there would be a place for other methods for stable fracture management besides on the one hand the plaster cast or pin osteosynthesis, respectively, external fixateur and, on the other hand, the traditional plate fixation (from a palmar or dorsal direction). PATIENTS AND METHODS: In this paper, we report our first experience with a novel plate that has been in use in Europe since April 2005. It is a fixed-angle, intrafocal nail plate. We started using it in April 2005 and up to October 31, 2006 have treated 32 patients. RESULTS, COMPLICATIONS: After one year we can only provide provisional results. Among the 32 cases we experienced two complications: a rupture of the long thumb extensor tendon and a loosening of the locking screws. CONCLUSIONS: The nail plate always has a firm place in our daily routine for those cases where a stable osteosynthesis with minimal impairment of soft tissue is desired. A prerequisite for success is a correct indication and an exactly performed operative technique. Apart from a palmar splint for ten days to spare the soft tissue, immobilisation is not necessary. PMID- 17402146 TI - [Candidemia must be treated!]. PMID- 17402147 TI - Knowledge is power. PMID- 17402148 TI - Beyond the internal. PMID- 17402149 TI - Pneumocystis carinii jiroveci pneumonia following infliximab infusion for Crohn disease: emphasis on prophylaxis. PMID- 17402150 TI - Lung cancer metastasis mimicking gluteal abscess. PMID- 17402151 TI - Do no harm to the arm!: hypertension and unilateral hand rash. PMID- 17402153 TI - World AIDS Day. PMID- 17402154 TI - Stronger leg muscles may thwart knee arthritis. PMID- 17402155 TI - Things that make you ache. The possibilities are many. PMID- 17402156 TI - I had surgery in 2005 to treat my enlarged prostate. The procedure was transurethral resection of the prostate. Things have worked out pretty well, but I'm wondering how long I can expect the improvement in urine flow to last? PMID- 17402157 TI - I tried a new weight-loss supplement that contains green tea extract. About an hour after I took the pill, my heart was pounding and seemed to be racing. Could this be from the green tea or something else in the pills? PMID- 17402159 TI - Tea with a twist. PMID- 17402158 TI - New Start! for exercise. PMID- 17402160 TI - Parkinson's drugs linked to heart valve trouble. PMID- 17402161 TI - Ask the doctor. My doctor put me on a statin and told me to take it after dinner. I would rather take it with breakfast. Does it matter? PMID- 17402162 TI - Poorly differentiated prostate cancer treated with radical prostatectomy: long term outcome and incidence of pathologic downgrading. PMID- 17402163 TI - Glycemic durability of monotherapy for diabetes. PMID- 17402164 TI - Glycemic durability of monotherapy for diabetes. PMID- 17402165 TI - Glycemic durability of monotherapy for diabetes. PMID- 17402166 TI - High-resolution endoscopy and early gastrointestinal cancer...dawn in the East. AB - Cancer in the esophagus, stomach, and large bowel accounted for 24 % of worldwide cancer mortality in 2002. This burden is expected to increase further as the world's population ages. Prevention is based on early detection of potentially curable cancers and/or precursor conditions that have a significant risk of progression to malignancy. Early detection in the asymptomatic population can be achieved in: (a) people who comply with interventions proposed by health authorities (population-based screening); (b) people with a link to a particular health policy (e. g. occupational screening); and (c) people who contact their own doctor or private health check-up institutions (individual or "opportunistic" screening). Factors that limit the benefit of early detection and treatment of neoplastic lesions include the degree of compliance with the selection procedure and compliance with recall for diagnosis and treatment in people with a positive test. The miss rate for small but potentially cancerous lesions is often responsible for interval cancers. There is also a danger of overdetection and overtreatment of lesions with a very low risk of progression to malignancy. PMID- 17402167 TI - A new laparoscopic technique: suction removal of spilled gallstones. PMID- 17402168 TI - Radiologic gastrostomy. AB - Radiologic gastrostomy has a higher success rate and a lower complication rate and offers a greater choice of tubes than percutaneous endoscopic gastrostomy (PEG). The position and configuration of the stomach and colon are clearly seen under fluoroscopy, and ultrasound can be used to locate the liver. Radiologic gastrostomy procedures can be performed when there are oropharyngeal tumors, or esophageal strictures and stents, and can be performed under local anesthesia alone. Peroral push-gastrostomies are preferable for palliative care and for patients with neurogenic dysphagia, but percutaneously inserted tubes should be used in patients with upper gastrointestinal cancers in order to avoid tumor seeding. Unfortunately, awareness of and access to radiologic techniques are still limited and this has led to the development of "adventurous" techniques for placing endoscopes in stomachs rather than applying simple fluoroscopic alternatives. PMID- 17402169 TI - Correction: Plasma exchange when myeloma presents as acute renal failure. PMID- 17402170 TI - Commentary. Early carotid endarterectomy after ischemic stroke: the results of a prospective multicenter Italian study. PMID- 17402171 TI - [Radiofrequency catheter ablation at left coronary cusp for left ventricular repetitive monomorphic tachycardia: report of one case]. PMID- 17402173 TI - The worm that turned. PMID- 17402172 TI - Health care human resources. PMID- 17402174 TI - Quantitative venom proteomics. PMID- 17402175 TI - A systematic future? Interview by Katherine Vickers. PMID- 17402176 TI - The soft cell. PMID- 17402177 TI - The Boxing boys of Akrotiri: early illustration of spondylolytic spondylolisthesis related to a sports injury. PMID- 17402178 TI - Where ribs can be found in odd places. PMID- 17402180 TI - The rise and rise of green chemistry in China. Interview by Nicola Nugent. PMID- 17402179 TI - The poor man's atomic bomb. PMID- 17402181 TI - A natural selection. PMID- 17402182 TI - [Changes in intestinal microflora following levofloxacin administration in laboratory animals]. AB - Administration of antimicrobial agent is the most frequent and significant cause of modifications to gastrointestinal bacterial microflora. The aim of the present study was to evaluate the impact on Gram-negative aerobic bacterial intestinal flora, the possible colonization of Candida spp. and the possible emergence of resistant bacterial strains in a ten rats group receiving levofloxacin at the dosage of 10mg/kg one a day for 5 consecutive days. Our result are in agreement with those of other studies and the reveal that Gram-negative aerobic bacteria are strongly suppressed during levofloxacin treatment. Neither colonization by Candida spp. nor emergence of resistant micro-organism to levofloxacin were observed. After 7 days from completing treatment, bacterial intestinal flora was restored in 8 out of 9 rats. Levofloxacin does not significantly affect intestinal microflora. Therefore, it may be suitable for selective decontamination of the gastrointestinal tract in immunocompromised patients, for prophylaxis of urinary tract infections and for treatment of intestinal bacterial infections. PMID- 17402183 TI - [Ethics and pediatric cardiopulmonary resuscitation]. AB - Cardiopulmonary resuscitation (CPR) is a medical activity that involves major ethical issues. As in other areas of clinical ethics, CPR decisions must be based on the principles of autonomy, beneficence, nonmaleficence, and justice. The decision-making process is more difficult in emergency situations, and when the patient is a minor, the parents and the child's best interests must be taken into consideration. There are specific situations in which starting CPR is clearly indicated and others in which ceasing resuscitation maneuvers is justified. Do not attempt resuscitation orders must be respected by health staff. Other ethical issues involved in CPR include resuscitation of potential organ donors, learning CPR procedures, research in CPR, and the information given to the parents of children with cardiorespiratory arrest. PMID- 17402184 TI - [Material for the pediatric resuscitation trolley]. AB - Cardiorespiratory arrest and the need for cardiopulmonary resuscitation can occur anywhere, both in the out-of-hospital and in-hospital settings. Therefore, all healthcare centers (hospitals, primary care facilities, out-of-hospital emergency services) must be prepared to initiate life support procedures in children and to treat other life-threatening emergencies. To achieve this objective, adequate material including a full crash cart or resuscitation trolley is essential and must be available in all healthcare centers. Specific items contained in the trolley can vary according to the characteristics of the facility and the most probable type of resuscitation needed (for example, neonatal resuscitation). At least one resuscitation trolley must be available in primary care centers, pediatric intensive care units, emergency departments, out-of-hospital emergency services, and pediatric wards. The trolley must be located in an easily accessible site and must contain only indispensable material. It is essential to include instruments in several sizes, covering children of all ages, as well as enough spare instruments and medications that could be required during resuscitation. The material must be checked periodically and all the staff (physicians, nurses, and auxiliary personnel) must be familiar with the trolley's contents and the location of all material and drugs. PMID- 17402185 TI - [New recommendations for uniform reporting of advanced life support]. AB - Pediatric patients requiring cardiopulmonary resuscitation show high morbidity and mortality. There are few studies on this topic and existing studies use distinct terminology and methodology in data collection, hampering comparisons, efficiency assessment, and meta-analyses, etc. Consequently, in clinical studies of cardiorespiratory arrest (CRA) and cardiopulmonary resuscitation (CPR) in the pediatric age group, data collection should be performed in a uniform manner. To define the criteria that allow uniform data collection, in 2004 a working group of the International Liaison Committee on Resuscitation published simplified recommendations for registering essential information, which could be applicable to adults and children both in clinical practice and research, as well as inside and outside the hospital setting. Following the Utstein style, the Spanish Group of Pediatric and Neonatal CPR has designed an algorithm and a data collection form for recording essential CPA data. The need for these documents to be designed with maximum accuracy is stressed, both because of their medico-legal and professional implications and because of the influence of some variables on post-CPA recovery. Likewise, while protecting patient confidentiality, provincial, regional and national CPA registries should be developed, which would improve the quality of care, research, and resource provision according to needs. PMID- 17402186 TI - Oklahoma Dental Association Centennial 1907-2007. PMID- 17402187 TI - A highly paradoxical historian. PMID- 17402188 TI - [Study on the socio-economic risk factors on birth defects in Lvliang district of Shanxi province]. AB - OBJECTIVE: To study the risk factors in high prevalence district of birth defects (BD) to provide evidence for intervention development. METHODS: Quantitative analysis had been carried out based on the investigation at three counties in Shanxi province. 491 useable questionnaires had been collected with a response rate as 94.6%. Binary logistic regression was performed to analyze the risk factors of BD. RESULTS: Data from logistic regression analyses revealed that factors as: the status of family income, knowledge on healthy birth and rearing, behavior and illness during pregnancy were influencing the occurrence of BD, with OR values as 0.535, 3.265, 0.403 and 1.379 respectively. Better family income and knowledge on BD were negatively (P values are 0.000 and 0.001 respectively), while illness during pregnancy and alcohol intake of the husbands were positively correlated (P values are 0.005 and 0.012 respectively) to the occurrence of BD. CONCLUSION: Measures as accelerating the development of local economy, providing education on BD knowledge to the couples, changing negative life style and improving the health status would reduce the risk of BD. Preconceptional periconceptional care seemed to be the new idea for healthy pregnancy and healthy baby. PMID- 17402189 TI - [Study on the quality of life and its influencing factors in Miao and Gelao Minority groups in Guizhou province]. AB - OBJECTIVE: To investigate and evaluate the quality of life and its influencing factors in Miao and Gelao Minority groups in Guizhou province with the aim of providing evidence for valid health promotion measures. METHODS: Based on a self designed questionnaire to evaluate the quality of life on 582 subjects. Kruskal wallis test and stepwise regression analysis were used to test the quality of life and its influencing factors. RESULTS: There were significant differences among the groups of quality of life and its influencing factors. The quality of life of Miao Minority was the highest and Gelao the lowest. Sex, age, acute disease prevalence of the past two-weeks, prevalence of chronic diseases, drinking unboiled water, tobacco-smoking habit, educational level and the family expenditure on life were correlated with all or most of the domains of quality of life. CONCLUSION: The key steps on quality of life promotion should include: strengthening the basis medical health service, pushing cooperative medical service, enhancing prevention and care of chronic diseases, speeding the development of economics etc. PMID- 17402190 TI - [Study on the baseline physical activity and the risk of ischemic cardiovascular diseases]. AB - OBJECTIVE: To examine the associations between physical activity measures [metabolic equivalents of energy expenditure (MET) per hour per day] and ischemic cardiovascular diseases (ICVD) in Chinese population. METHODS: A survey on cardiovascular risk factors was conducted in different areas of China in fall 1998. People aged 35 to 59 but without a history of coronary heart and stroke at baseline were prospectively followed and 11 849 subjects whose information were complete at the end of second follow-up were valid. Cox regression was used to estimate the hazard ratios (HRs) for incident ischemic cardiovascular diseases and the different measures of physical activity. RESULTS: During a mean follow-up period of 5.9 years, 84 incident ischemic cardiovascular events were ascertained. We examined the HRs of ischemic cardiovascular events for a 1-unit change in METs value, which were included in the models as continuous variable. There were negative association of METs values found with ischemic cardiovascular events in total, urban, rural, male and female subjects, and statistical significance in the urban (HRs = 0.22, 95% CI: 0.05-0.95) but the association was weakened after adjustment for demographic factors. When further adjustment for other intermediate factors, the significance in the urban was again attenuated. When the urban males and females, rural males and females were divided into 3 groups according to their respective tertiles and the combination of different population groups, the factors of male/female and urban/ rural were equally distributed in different groups, and no more adjustment in the Cox model. The multivariate - adjusted (age and education attainment) HRs associated with the tertiles, from lowest to highest, were: 1, 1.03 and 0.65 (P(trend) = 0.170) for the total, 1, 0.72 and 0.64 for the urban, 1, 1.49 and 0.72 for the rural, 1, 1.05 and 0.59 for men, 1, 0.90 and 0.84 for women. CONCLUSION: The totality of our findings pointed to METs per hour per day seemed to be weakly associated with a reduction in ischemic cardiovascular events incidence among urban middle-aged adults. PMID- 17402191 TI - [Study on the equity of rural health service in the experimental region of new rural cooperative medical scheme, Yuexi county, Anhui province]. AB - OBJECTIVE: To explore the situation of equity in health service delivery in the experimental region of new rural cooperative medical scheme (NRCMS). METHODS: A household questionnaire survey was conducted to 2112 residents in the experimental county of NRCMS in Yuexi county with a stratified-cluster sampling on the situation of health service delivery, from July 1-15,2005. Data was analyzed with Epi Info 6.04 and SPSS 11.0 software and indices as rate, ratio, Gini coefficient, concentration index (CI) and chi-square test etc. RESULTS: There was no significante difference of the treatment-seeking rate during two weeks among five groups economic condition residents (chi2 = 5.52, P > 0.05) and the rates were 48.14 per thousand, 82.90 per thousand, 65.88 per thousand, 48.72 per thousand and 50.66 per thousand respectively with CI = -0.026. Similarly, the hospitalization rates were 59.08 per thousand, 67.36 per thousand, 51.76 per thousand, 58.97 per thousand and 52.86 per thousand respectively in the last year and the CI = - 0.017. But there was a significante difference of rates on chronic disease among the five groups of residents with different economic conditions in the last six months (chi2 = 18.42, P < 0.01) and the rates were 295.40 per thousand, 243.52 per thousand, 230.59 per thousand, 225.64 per thousand and 176.21 per thousand and the CI = -0.055. When income reduced, the prevalence had been increasing among residents. Meanwhile, the collection funds showed unfair in residents with various income and the compensating fund of new rural cooperative medical scheme had not reduced the gap between rich and poor. CONCLUSION: There was an unequity of medicine expenditure and compensating fund in residents with various income in the experimental region. The low income residents had a high health service need and the government should improve NRCMS greatly to change the situation. PMID- 17402192 TI - [Incidence rates of human immunodeficiency virus and syphilis as well as the rate of retention in a 6-month follow-up study of female sex workers in areas with heavy drug use in Xichang of Sichuan province, China]. AB - OBJECTIVE: To investigate the human immunodeficiency virus (HIV) and syphilis incidence rates as well as the retention rate in a cohort with 6-month follow-up study among female sex workers (FSWs). METHODS: From December, 2004, to January, 2005, a community-based baseline survey was conducted to recruit 343 FSWs for a prospective cohort study in Xichang county of Sichuan province, China. Follow-up visits were conducted at 6 months to analyze risk factors associated with cohort retention for subjects' baseline sociodemographic and sexual behavioral characteristics. Blood specimens were also collected to test antibodies against HIV and syphilis. RESULTS: During the 6-month follow-up period, HIV and syphilis incidence appeared to be 1.00 per 100 person-years and 6.23 per 100 person-years, respectively. The rate of retention in the cohort was 53.6% (184/343). Results from multivariate logistic regression model showed that factors were significantly associated with cohort retention including people with minority ethnic background (OR = 0.36; 95% CI: 0.18-0.74), people having participated in AIDS prevention program (OR = 1.83; 95% CI: 1.17-2.86) or being clients in the last 6 months > or = 50 (OR = 1.75; 95% CI: 1.11-2.77) and having changed living/working place (OR = 0.56; 95% CI: 0.33-0.94). CONCLUSION: The results of this study showed that the syphilis incidence and unprotected sex behavior were high among local FSWs. People belonged to Han nationality, having participated in AIDS prevention program and having a steady living/working place were associated with cohort retention at 6-month follow-up study among FSWs, respectively. PMID- 17402193 TI - [Study on the status of pollution by Legionella species in hot spring vacation center and the related factors]. AB - OBJECTIVE: To study the pollution status of Legionella species in hot spring vacation center and the related factors. METHODS: Field surveys were performed in four big hot spring vacation centers of Changping district. Uniform questionnaires was used and colony count was made together with the isolation of Legionella species from hot spring water based on mip gene typing. RESULTS: 47 isolates of Legionella pneumophila (Lp) from 87 samples showed 4 serotypes as Lp1, Lp6, Lp12, Lp5 with percent of 57.45%, 21.28%, 14.89%, 6.38% respectively. The hot spring centers controlled the temperature of recycled water between 34-47 degrees C by hot water heating and filtrating system. All the isolates were cultured from the hot water with temperature between 34-44 degrees C: 56.75% (21/37) in high temperature (40-47 degrees C) and 61.90% (26/42) in low temperature (34-39.9 degrees C). There were no statistically significant difference between the high and the low temperature samples (P > 0.05). In the four hot spring vacation centers, the pH value was under control at 6.4-7.3 and the ambient temperature was under control between 26-28 degrees C. The humidity was controlled between 56% -69% relative humidity, which were the best growing conditions for the Legionella species. Disinfectors as chlorine deviratives was used in the four hot spring vacation centers. Though the concentration of chlorine in the water was 0.3-0.5 mg/L, 14.29%-48.00% of the samples were still positive of having Legionella species. CONCLUSION: The pollution of Legionella species was considered to be quite serious in the four hot spring vacation centers and the predominant serotype was Lp1. The pH and temperature of the hot spring water, ambient temperature and humidity and the way of heating up the water were the best conditions for the growth of Legionella species in these centers. Because of the high temperature of the hot spring water, chlorine of the disinfector volatilized quickly, affecting the effect of disinfection. The result revealed that water temperature achieving 44 degrees C could have had the effect of prevention. PMID- 17402194 TI - [Study on the mixed testing of serum samples in seroepidemiological survey of parasitic diseases]. AB - OBJECTIVE: In order to accumulate experiences for improving the efficiency in serological tests, the present study on mixed testing of serum samples was performed by taking the serological test of trichinellosis and toxoplasmosis as the examples, and had proved the effects on cost-effectiveness of seroepidemiological survey of parasitic disease with method of mixed-samples test. METHODS: According to the binomial distribution principle, to develop an approach to the feasibility of mixed testing of serum samples, and to work on a cost-effectiveness analysis of one-by-one testing and mixed testing using hygienic economic analysis method was performed. For serological test of trichinellosis and toxoplasmosis, 3 kinds of mixed testing methods, namely 3 serum sample mixture, 5 serum sample mixture and 10 serum sample mixture, were performed. RESULTS: The results showed that all the 3 kinds of mixed tests of trichinellosis and toxoplasmosis showing positive result if only 1 weak positive serum sample were mixed with. When the serum samples being mixed were all negative ones, then among the 24 groups tested with each kind of negative serum sample mixture of trichinellosis (3 serum samples, 5 serum samples and 10 serum samples), they all showed negative. However, among the 12 groups tested with 2 kinds of negative serum mixture of toxoplasmosis (3 serum samples and 5 serum samples), all showed negative while among the 18 groups tested with the 10 serum sample mixture, 16 groups showed negative and 2 were positive. The mixed testing of trichinellosis and toxoplasmosis showed that the efficiency of mixed testing was related to the serological positive rate of the parasitic diseases to be examined. When serological positive rate was 10%, the efficiency of mixed testing was higher in 4 serum sample group. When serological positive rate was 1%, the efficiency of mixed testing was higher in 10 serum sample group and when serological positive rate was 0.1%, the in crease of the size of mixed serum samples could decrease the number of testing, but the prerequisite was that there must be one positive sample, so that the positivity for all the mixed tests could be detected. If mixed testing were performed on all negative samples, no positivity could be detected. CONCLUSION: The result of cost-effectiveness analysis demonstrated that for seroepidemiological survey of parasitic diseases, the cost for mixed testing was low, especially when the serological positive rate was expected low (< or = 1%, thus the mixed testing could save a large amount of the cost. PMID- 17402195 TI - [Study on the dynamic prevalence of serum antibody against severe acute respiratory syndrome coronavirus in employees from wild animal market in Guangzhou]. AB - OBJECTIVE: To investigate the dynamic trend of specific antibody against severe acute respiratory syndrome (SARS)-CoV in serum collected at various periods among employees in Guangzhou Xinyuan animal market. METHODS: Volunteers from employees of the animal market were recruited and their serum specific antibody against SARS-CoV were determined by enzyme linked immunesorbent assay (ELISA) method. RESULTS: Positive SARS-CoV specific IgG antibody was found 25.61% (n = 328), 13.03% (n = 238), 12.59% (n = 135), 5.04% (n = 139) and 9.43% (n = 53) among volunteers, which were sampled in May 2003, Dec. 2003, Jan. 2004, July 2004 and June 2005 respectively. No specific IgM antibody was found in all of those samples. Among 129 samples which were tested twice or more, 97 were all negative, 18 all positive, 13 changed from positive to negative but only one sample from negative to positive. When the volunteers were divided by the duration of their working experiences as short-term or long-term, those who had worked at animal market for less than or more then 6 months when being tested, the positive rate for long-term employees were relatively constant, however, all of the persons employed after January 2004, when the palm civets and raccoon dogs were culled from the market, were tested negative. CONCLUSION: The prevalence of specific antibody against SARS-CoV in employees of the animal market were somehow related with the presence or absence of palm civet. No serum was tested positive for persons who were employed after palm civets and raccoon dogs were culled from market. This data indicated that the SARS-CoV might have been from the palm civets and raccoon dog, and the animal market seemed to serve as one of the sources of infection. PMID- 17402196 TI - [Study on human case of avian influenza in Guangzhou 2006 without causing human to-human transmission among close contacts]. AB - Study on human case of avian influenza in Guangzhou 2006 without causing human-to human transmission OBJECTIVE: To explore the possibility of transmission from a human case of avian influenza to his close contacts. METHODS: Close contacts of the human case of avian influenza in Guangzhou 2006 were found out according to the definition and methods publicized by the Ministry of Health, People's Republic of China. Epidemiological investigation and medical observation were carried out. Serum antibodies were tested in some of the close contacts. RESULTS: The avian influenza patient had never left Guangzhou in the month prior to disease onset. No contact history with dead or diseased poultry was found. A total of 56 close contacts, including his girl friend, relatives, friends and medical staff who had taken care of him, were brought under medical observation for 7 days but none of them showed signs of infection. CONCLUSION: Unlike SARS, direct contact with patient contracted with avian influenza at the end of incubation period and in the stage of illness through flying droplets, saliva, mucous membrane and skin injuries will not lead to human-to-human transmission, indicating the virus' ability to pass from human to human is limited. PMID- 17402197 TI - [Analysis on the factors related to rabies epidemic in China, in 2005]. AB - OBJECTIVE: To analyse the related epidemic factors of rabies and to provide basic evidence for developing rabies control and prevention programs in China. METHODS: Possible factors that causing rabies epidemics in the southern part of China were obtained through collecting both epidemic and surveillance data of rabies in 2005, and to analyse every factor by software of Excel 2002. RESULTS: The number of rabies cases was 2548 in 2005. Among 885 cases being investigated, 60.56%, 49.04%, 96.16% had not had any chance to receive treatment, vaccine or immunoglobulin. Among the exposed persons, 89.95% received treatment on wounds and 94.93% were immunized, and 8.56% received immunoglobulin. 94.86% of them accomplished the full rabies post-exposure prophylaxis course, but 8.56% of them did not accomplish it. 88.50% of the cases and 92% of the persons under exposure were bitten by dogs. The density of dogs was 3.20-13.37 per 100 persons. The vaccination coverage rates among dogs were 5.31% -75.11% with a positive rate of 2.93 % -6.40%. CONCLUSION: Factors as: low rate of post-exposure prophylaxis and failed to have finished the necessary post-exposure treatment (PET) of the cases, nonstandard PET in hospital or local clinic, low rabies vaccination coverage and high infectivity of rabies virus in animal host (mainly for dog) might be responsible for rabies epidemics in China, in 2005. PMID- 17402198 TI - [Estimation on the level of birth and death rates of population in the three gorges area by means of capture-mark-recapture method]. AB - OBJECTIVE: To evaluate quality of surveillance and emendate rates of birth and death of population of the Three Gorges area. METHODS: Data on the two samples collected were designed based on principle of capture-recapture method. An investigation of missing report of birth and death was conducted in 7061 families selected through stratified random sampling method. We collected and registered the data of birth and death in every family investigated and checked with correlative records reported in disease surveillance system of the Three Gorges area. The missing report rates and the 95% confidence intervals of birth rate and death rate were calculated. RESULTS: The underreporting rates of birth and death were 13.91% and 15.60% and death of infant was 33.33%. The emended birth rate was 8.92 per thousandth and the 95% confidence interval of birth rate was 8.38 per thousandth-9.45 per thousandth. The emended report rate of death was 6.88 per thousandth and the collectivity 95% confidence interval was 6.37%-7.38 per thousandth. CONCLUSION: Results showed that the quality of birth and death in the disease surveillance reporting system of Three Gorges area was competent to the quality level of the standard set for national disease surveillance system. The birth and death rates of population in the Three Gorges area were under 10.00 per thousandth. PMID- 17402199 TI - [Detection of Rickettsia prowazekii by quantitative real-time PCR]. AB - OBJECTIVE: To develop a quantitative real-time polymerase chain reaction (PCR) for detecting Rickettsia prowazekii. METHODS: Primers and TaqMan-MGB probes designed based on ompB gene of R. prowazekii, were used to develop this method. RESULTS: For the quantitative real-time PCR, the relationship between the values of threshold cycle (Ct) and the DNA copy number was linear (r = 0.999) and the sensitivity was about 100 times higher than that of the nested PCR for detecting the same DNA sample. The results of the genomic DNA samples of other rickettsial and bacterial agents detected by real-time PCR were all negative. DNAs extracted from blood samples of guinea pig infected with R. prowazekii were examined by real-time PCR and the positive results were obtained from some of these samples. However, the results of some samples in nested PCR assay were all negative. CONCLUSION: These results suggested that the real-time PCR was highly specific and sensitive for detection of R. prowazekii that was useful for the detection of tiny DNA of R. prowazekii in blood samples from patients suspected of having epidemic typhus. PMID- 17402200 TI - [Study on the relationship between the polymorphisms and secondary structure of tat exon-1 gene and HIV/ AIDS progress in subtype B' and B'/C]. AB - OBJECTIVE: To study the polymorphisms and secondary structure of human immunodeficiency virus (HIV-1) tat exon 1 among subtype B' and B'/C HIV-1 infected people in China and to explore the relationship between the polymorphism of tat exon 1 and the disease progression. METHODS: 8 subtype B' and 5 B'/C HIV-1 infected patients with slow disease progression were selected from Liaoning, Jilin and Yunnan province. 26 subtype B' and 9 B'/C HIV-1 infected patients with similar sex, age but with typical disease progression were selected. Provirus was extracted from the whole blood. The gene sequences of the Tat exon 1 were amplified by nest-polymerase chain reaction (nest-PCR). Products were purified and sequenced directly. The sequences were aligned, translated, amino acid substitution were analyzed and secondary structures were predicted. RESULTS: Many amino acid substitution could be found in the exon 1 of Tat in HIV-1 subtype B' and B'/C recombinant strain infected persons with different disease progression except A58T,none of them showed definitely relationship with HIV viral load and disease progression. 23N, 31S, 32Y and 46F were subtype-specific substitutions. No characteristic secondary structure of exon 1 of Tat was found. CONCLUSION: Some of the mutations of tat exon 1 might be related to HIV viral load and disease progression. However, there was no relationship found between the secondary structure of Tat protein and the disease progression. PMID- 17402201 TI - [Study on the characteristics of mutation on Mycobacterium tuberculosis rifampicin-resistance gene]. AB - OBJECTIVE: To elucidate the characters of rpoB mutation in rifampin-resistant clinical isolates of Mycobacterium tuberculosis. METHODS: 286 bp DNA fragment of rpoB gene including 81 bp code region (rifampin resistance deteremination region, RRDR) was analyzed by PCR-single-strand conformation polymorphism(SSCP). The 286 bp DNA fragment of each strain which had been proved to have mutation by PCR-SSCP was then sequenced. 110 strains of M. tuberculosis, including 73 rifampin resistant strains, 11 rifampin-susceptible drug-resistant strains and 26 drug susceptible strains were studied. RESULTS: 47 rifampin-resistant strains were detected to have mutations by PCR-SSCP method. 76.6% rifampin-resistant strains showed that rpoB gene was carrying single point mutation analyzed by direct sequencing technique, which mainly located at 531-Ser (61.1%) and 526-His (25.0%). The combined mutation rate was 23.4%. In addition, 2 rifampin susceptible drug-resistant strains and 1 drug-susceptible strain were mutated, detected by PCR-SSCP method. Sequencing results showed that the mutations located at 511-Leu, 526-His and 535-Pro. CONCLUSION: Mutations in the 81 bp RRDR of rpoB were the main reasons of M. tuberculosis resistant to rifampin. 531-Ser and 526 His were the most common positions of mutations. PMID- 17402202 TI - [Investigation on virus genotype in patients infected with hepatitis B virus in four cities of Guizhou]. AB - OBJECTIVE: To investigate the distribution of hepatitis B virus (HBV) genotype in Guizhou and to study the relationship between the genotype and the progression of liver disease. METHODS: 786 patients with chronic HBV infection, from 4 cities of Guizhou, including 346 asymptomatic carriers (ASC), 313 chronic hepatitis (CH), 77 liver cirrhosis (LC), 50 hepatocellular carcinoma (HCC) were examined. HBV genotype was determined by restriction fragment length polymorphism analysis and the subtypes were determined by direct sequencing of PCR product in 94 patients with HBV B genotype, the relationship between HBV genotype and the progression of liver disease was studied by multifactor analysis such as HBeAg positivity, HBV DNA load and ALT level. RESULTS: Of the 786 patients, 7 (0.89%), 497 (63.23%), 275 (34.99%), and 7 (0.89%) belonged to genotype A, B, C, D, respectively. There was statistically significant difference in the distribution of genotype B among Kaili (96.04%), Zunyi (78.79%), Duyun (64.52%) and Guiyang (53.14%) (P< 0.01). Genotype C was more prevalent in Guiyang than in other three cities (P < 0.01, or P < 0.05). Out of 94 genotypes B, 93 (98.94%) belonged to subtype Ba, only one was subtype Bj. There were statistically significant difference in the distribution of genotype B and C among various stage of liver disease (P < 0.05 or P < 0.01). Genotype B showed a gradual decrease from ASC, CH, LC to the HCC group while in contrast, genotype C showed a gradual increase in the same order. The ALT levels and the mean age were significantly higher and older in patients with genotype C than those in genotype B (P < 0.01 or 0.05). The HBeAg positivity was significantly lower in genotype C than that in genotype B (P < 0.025). CONCLUSION: Data showed that there were genotype A, B, C and D existing in Guizhou. Genotype B was the major one but genotype C was more commonly seen. In genotype B, subtype Ba appeared to be predominant. The geographic distribution of genotype B and C were different in some cities of Guizhou. Compared to genotype B, genotype C was associated with the development of more severe liver damage. PMID- 17402203 TI - [Surveillance on natural infection of rodents with hantavirus in Shenzhen city and identification of a hantavirus strain SZ2083]. AB - OBJECTIVE: For clarifying the situation of the natural infection of rodents having hemorrhagic fever with renal syndrome (HFRS) virus and to type Hantavirus (HV) using molecular technique in Shenzhen city in 2005, and offering guidance for prevention and control of HFRS. METHODS: Data on the host animals was collected from the city of Shenzhen. ELISA and indirect immunofluorscent antibody(IFA) test were applied to the specific antibodies against HV in the sera of captured rats. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues of the HV infected rats were inoculated into Meriones unguiculata to isolate HV. The whole viral RNA was extracted from the lung tissues of the HV infected rats and amplified the partial M fragments with RT-nested-PCR, using the HV genotype specific primers. The amplified genes were then sequenced, and subjected to genotyping and homology analysis. RESULTS: 472 rodents were captured from Shenzhen in 2005. Surveillance on rats demonstrated 9.96% rats carrying HV (with a density of 8.25%) and the main host was Rattus norvegicus. In the blood samples of rats, anti-HV IgG antibodies were detectable in 56 cases by IFA, and proved to be positive in 76 cases by ELISA. We successfully isolated a HV strain designated as SZ2083 from Rattus norvegicus for the first time in Shenzhen and was identified to SEO type by RT-nested-PCR. Compared with the coding region of the M gene of HV L99 virus strain, the homologies of nucleotide among them were 97%, but the homology was 76% of the SZ2083 with HTN 76-118 virus strain. CONCLUSION: Results showed the existence of natural epidemic areas of HFRS in Shenzhen city. Based on the results of sequencing, it is possible that the Seoul strain of HV might be the predominant serotype of virus harbored. PMID- 17402204 TI - [Evaluation on the effect of intervention regarding breast self-examination for decreasing breast cancer mortality]. AB - OBJECTIVE: A randomized trial of breast self-examination (BSE) Program was carried out to evaluate whether the intensive BSE could reduce the number of deaths among women from breast cancer. METHODS: This study was a randomized controlled trial (RCT). A total of 266 064 women (age of 30 to 64 years) associated with 519 textile factories in Shanghai had been randomly assigned to a BSE group (132 979 women) or a control group (133 085 women) since 1989. Initial instruction in BSE group would include demonstration of proper palpation techniques and was followed by 2 reinforcement sessions during the subsequent 4 years including video shows, BSE instruction sessions and BSE practice under medical supervision. These activities were continued for 5 years. Attendance at all events was recorded. The cohort was followed through July 2000 for development of breast diseases, and the breast cancer cases were followed through 2001 for vital status. Data analysis methods used would include Kaplan-Meier plots, log-rank test and Cox modeling. RESULTS: Among women under instruction, 864 breast cancers detected and 133 breast cancer deaths occurred while 896 breast cancers were detected and 130 deaths recorded in the control group. The tumor size (P = 0.07), TNM stage (P = 0.39) and cumulative breast cancer mortality rate (P = 0.72) were not significantly different between the 2 groups. However, more and smaller fibroadenomas were detected in the instruction group than in the control group (P< 0.01). CONCLUSION: Intensive instruction in BSE did not seem to have reduced the mortality rate of breast cancer, but more and smaller benign breast lumps could be detected. PMID- 17402205 TI - [Study on the therapeutic status of 1242 hospitalized acute myocardial infarction patients in Beijing]. AB - OBJECTIVE: To investigate and analyze the therapeutic level and the existing problems on acute myocardial infarction (AMI) management in Beijing. METHODS: We collected clinic data of 1242 AMI patients from 12 hospitals in Beijing, from January 2000 to March 2001, using a uniformed questionnaire, and evaluated the status of the diagnosis and treatment of AMI according to the Chinese guidelines issued on Decmeber 2001. Corresponding factors which influencing the mortality were also analyzed by one-way factor and multiple factors analysis methodologies. RESULTS: The mean age of the 1242 AMI patients was 63.0 years old and about one third of them were under 55 years old. In hospitals, the total mortality was 9.10%. 37.9% of the patients had received therapy of the intravenous thrombolysis and emergency PCI with a total rate of reperfusion therapy as 56.0%. The in hospital rates of drug use were as follows: Nitrates 90.0%, Aspirin 87.8%, heparin 88.7%, beta-blockers 73.4%, angiotensin converting enzyme inhibitors(ACEI) 77.6%, lipid regulating agents 43.6%. The rate of intravenous therapy of TCM by promoting the blood circulation and supplementing the vital energy was 30.5%. Results from multiple factors analysis showed that the compositive factors which could lower the mortality were reperfusion therapy,lipid regulating agents, intravenous therapy of TCM, beta-blockers, ACEI, lower molecule heparin and digitalis. CONCLUSION: Data from this study showed that there still existed a gap between clinical management on AMI and the guideline in Beijing. To set up a straightway passage of reperfusion therapy, to become more standardized to follow the guideline in undertaking the medical treatment practice, and to go deep into discuss the status of TCM on AMI management seemed the important tasks we are facing. PMID- 17402206 TI - [Study on prehospital time and influencing factors of stroke patients in 15 Chinese cities]. AB - OBJECTIVE: To study the prehospital time and influence factors of stroke patients in 35 hospitals from 15 cities. METHODS: Unified questionnaires were used to investigate all of the 2270 stroke patients after at the first week of onset from June 30, 2002 to April 30, 2003. Other than general data, the patients were divided into two groups according to prehospital time within or after 6 hours and all the factors influencing prehospital time of stroke patients were analysed with Chi-Square test and logistic regression. RESULTS: (1) All patients aged 18 102 with mean 64.8 y +/- 11.9 y including 60.2% of males. 78.8% of the patients were diagnosed as ischemic stroke and 21.2% hemorrhagic stroke. (2) 27.8% of the patients arriving at hospital immediately after the onset of stroke and 27.2% made emergency calls for ambulance. Only 25% of the patients had the sense that they should go to the hospital and receiving thrombolysis therapy after the onset of symptoms. 25% of the patients were ignorant of any knowledge about stroke, did not ask for help but waiting. (3) 57.5% of the patients arriving in hospital < or =6 hours after the onset and 58.6% went to emergency room directly. Thrombylysis rate among ischemic stroke patients arrived in hospitals < or = 6 hours was 6.7%. Multivariate logistic regression analysis showed following factors were related to arriving in hospital within 6 hours with P < 0.001: whether to emergency room directly, to hospital by ambulance or not, distance between stroke onset site to hospital, whether patients know thrombolysis was immediately needed after the onset of stroke, whether more attention to be paid on stroke after the onset of symptoms, whether needs to go to hospital at once after the onset ect. CONCLUSION: The main reasons for 42.5% stroke patients who arrived hospital over 6 hours were: lack of stroke knowledge, less using emergency call/ambulance,over 20 km between the site where stroke onset was occured and the hospital etc. To enhance public awareness about stroke was important to shorten the patients' arriving hospital's time. PMID- 17402207 TI - [Study on the diagnosis, treatment and requirement of epilepsy patients in urban communities]. AB - OBJECTIVE: To investigate the diagnosis,treatment and requirement of epilepsy patients in some urban communities in China, and to provide the evidence of searching for effective treatment and management on epilepsy under the China's context. METHODS: A face-to-face survey were conducted in 3 urban communities in Shanghai, Beijing and Changsha, respectively. The questions in the questionniare were general information, hospital visit, treatment, the level and way of getting on the knowledge of epilepsy, as well as the current obstacles and needs. RESULTS: Most of the patients selected the regular hospitals (90.8%) and the departments (92.3%) for their epielspy diagnosis and treatment. They used AEDs modo dictu (77.4%), and had controlled the seizures quite well (82.6%). A small part of patients still could not deeply understand the basic knowledge on epilepsy (13.5%). They ignored to follow up the drug concentration (45.8%) in blood and the blood biocheminstry indicators (43.9%). Some patients went to private clinics (12.9%) and used lay people remedies (7.7%). Longtime waiting (36.8%), inconvenient traffic (23.2%), and high expenses (22.6%) were the main problems influencing the timely treatment. The main obstacles of the patients were employment (47.2%), marriage (29.9%), psychological conditions (44.4%) and interpersonal relationship (29.9%). The main requirements were the effectiveness (87.0%) and cheap AEDs (40.9%) as well as the convenience of hospital visit (37.0%). CONCLUSION: It is very important to emphasize knowledge and publicity/education on epilepsy as well as the psychological treatment according to the requirements of patients. PMID- 17402208 TI - [Is the right ventricle able to maintain the systemic circulation for a long time? The late results of the Senning operation for complete transposition of the great arteries]. AB - The successful surgical management of congenital heart defects increased the number of such a type of defects, in which the morphological right ventricle has to provide the systemic circulation. Children with transposition of the great arteries operated with Senning procedure are good examples to investigate the long-term right ventricular function. Aim of our study was to determine the adequate non-invasive methods to assess the progression of the right ventricular dysfunction, to analyse the long-term surgical results and to determine the indication of reoperation. METHODS: Physical examination, electrocardiogram, 24 hours ECG, stress-testing, echocardiography, B-natriuretic peptide level measurement and magnetic resonance imagine were performed. PATIENTS: 176 Senning operations were carried out between 1980 and 1997. The follow-up period was 9-25 (mean 12.5) years and 83 children were included in our study. RESULTS: Significant correlations were found between the MRI RVEF, and echocardiographic TEI index, TDI and BNP results. Abnormal values were found in Senning patients compared to healthy children in MRI RVEF 55%, echocardiographic TEI index 0.33, in RV TDI systolic velocities 0.2 m/s, and B-natriuretic peptide levels 150 pg/ml. That means that right ventricular disfunction is expected in Senning children even without clinical symptoms. CONCLUSIONS: (1) BNP and echocardiographic TEI and TDI became available methods for the follow-up of right ventricular disfunction, and annual MRI is recommended. (2) Conversion of Senning to anatomic repair is considerable in selected young patients because the capacity of myocardium to respond to pressure overload decreasing with age. In older children and adults very close follow-up is necessary using biomarkers, echocardiography and MRI to decide the correct time of heart transplantation. PMID- 17402209 TI - [Management of malignant and benign airway stenosis by stent implantation]. AB - INTRODUCTION: In the last few decades the different bronchoscopic procedures have gained an important role in the treatment of airway stenosis, and the number of implanted airway stents has also greatly increased. PATIENTS: Between 1998 and 2004 the authors implanted altogether 108 airway prosthesis in 90 patients at the Institute of Pulmonology of Pest County. 58% of the patients were males, 42% females, the average age was 57.5 years, the average follow-up time was 7 months. RESULTS: On the basis of different etiology the patients were separated into two main groups. In 57% the airway stenosis was caused by malignant illnesses, in these cases stents can be used only with palliative purpose. However, in case of benign lesions they can offer a long-term solution and require an adequate follow up of the patients. The authors' main aim was to get an overall picture of the interventions they had done by processing the data, with the help of the measurable characteristics that make possible to follow the airways' permeability and its changes. Analysing the results of the respiratory function and blood gas examinations they didn't find a significant difference inspite of the subjective improvement. PMID- 17402210 TI - [The outcome of triplet pregnancies at the First Department of Obstetrics and Gynecology, Semmelweis University, in the last 15 years]. AB - INTRODUCTION AND METHODS: The authors analysed the data of 307 live-birth newborns and premature infants who were born from 104 triplet pregnancies between 1 January 1990 and 31 December 2005 at the Semmelweis University First Department of Obstetrics and Gynecology. AIMS AND RESULTS: In 86% of the triplet pregnancies conception took place with the help of assisted reproduction technology, the number of which has rapidly increased in the last 15 years. 60% of the triplet pregnancies ended before the 34th week of gestation and the birth weight of more than one third of the infants was less than 1,500 grams. Among the infants treated in the Neonatal Intensive Care Unit the most common diagnosis was connatal pneumonia, which was followed by hyaline membrane disease due to the low birth weight. In the past 15 years, the mortality of triplets was at 6.5%. The birth weight of those who died before reaching the age of six months was, with two exception cases, below 1500 grams. CONCLUSION: According to our data the chance of survival of triplets was poorer in category of birth weight under 1000 grams than those of premature infants born of single pregnancies with the same weight. In vitro fertilisation in the case of triplets increases the occurrence of premature deliveries and the number of infants with very low weight. Long-term monitoring data reveals that in such cases, neurological lesions and the occurrence of a chronic disease, which greatly influences the future quality of life for the child as well as for its family, is more frequent between the very low birth weight infants. PMID- 17402211 TI - [Novel method in diagnosis of chronic myeloproliferative disorders--detection of JAK2 mutation]. AB - Chronic myeloproliferative disorders are clonal hematopoietic stem cell disorders characterized by proliferation of one or more myeloid cell lineages in the bone marrow. The WHO classification describes six major groups of chronic myeloproliferative disorders, as follows: chronic myeloid leukemia, chronic neutrophilic leukemia, chronic eosinophilic leukemia, polycythemia vera, essential thrombocythemia and chronic idiopathic myelofibrosis. The diagnosis of chronic myeloid leukemia and certain types of chronic eosinophilic leukemia are based on the detection of fusion genes (in chronic myeloid leukemia the BCR/ABL fusion gene, and in chronic eosinophilic leukemia the FIP1L1-PDGFRalpha gene). On the other hand molecular markers for polycythemia vera, essential thrombocythemia and chronic idiopathic myelofibrosis were lacking, making it difficult to identify these disorders clearly. The authors investigated the incidence of the newly identified somatic point mutation V617F of the Janus-2 tyrosine kinase in patients with polycythemia vera, essential thrombocythemia and myelofibrosis. Janus-2 kinase is a cytoplasmic, non-receptor protein-tyrosine kinase with a key role in signal transduction from multiple hematopoietic growth factor receptors. The mutant protein is constitutively phosphorylated and is able to activate its downstream signaling pathways in the absence of any cytokine, thereby contributing to the pathogenesis of chronic myeloproliferative disorders. The authors investigated DNA samples from 132 patients with chronic myeloproliferative disorders. The V617F mutation was detected by allele-specific polymerase chain reaction, and the patients were genotyped by a DNA tetra-primer amplification refractory mutation system assay. Approximately 73% of polycythemia vera, 60% of essential thrombocythemia and 67% of myelofibrosis showed the JAK2 V617F mutation. Using the amplification refractory mutation system assay, the frequency of homozygotes was 17.5% in polycythemia vera, 5.4% in essential thrombocythemia and 0% in myelofibrosis. The authors established an effective polymerase chain reaction based method for the identification of JAK2 mutation in the routine oncohematologic diagnostics. PMID- 17402212 TI - [Hair analysis of abused and therapeutic drugs in forensic toxicology]. AB - Hair analysis for abused drugs has been gaining increasing significance in forensic sciences. Hair is a special matrix for the retrospective investigation of chronic drug abuse or poisoning in criminal cases and allows to demonstrate with sensitive methods even a single administration in low amount. Segmental hair analysis can yield the information about the time course of the substance use. The background of drug incorporation mechanism is not yet understood in full details and cannot be evaluated exactly in all cases. The hair sampling, sample preparation, analytical performance are very important for final results. The outcomes of hair analysis have been reviewed by dividing into six groups: opiates, cocaine, amphetamines, cannabinoids, abused therapeutic drugs and the markers of chronic alcohol consumption. PMID- 17402213 TI - [Laparoscopic transhiatal resection of epiphrenic diverticulum of the esophagus]. AB - The operation of the epiphrenic diverticulum of the esophagus--due to its anatomical position--is feasible either with thoracotomy or with laparotomy. The advantages of the first method are the possibility of better exploration and a technically easier operation. At the same time, the disadvantage of the surgical procedure imposed on the patient by opening his/her thoracic cavity is not to be underestimated, either. In case of an abdominal approach, the diverticulum in the mediastinum causes difficulties. The laparoscopic method combines the advantages of both solutions. The authors report the case of a 62 years old woman with a history of regurgitation and swallowing problems accompanied by a 20-kilogram weight loss. Barium esophagography and esophagogastroduodenoscopy showed a typically positioned 11-cm large epiphrenic diverticulum. As an operative solution transhiatal laparoscopic resection of the diverticulum with Heller cardiomyotomy and Dor fundoplication using an endoscopic stapler was performed. The postoperative period was uneventful; the patient was discharged on the 8th post-operative day with unhindered swallow. In the international literature 79 cases treated with minimally invasive therapy have been published so far. Based on own experience, the authors state that the laparoscopic transhiatal resection of the epiphrenic diverticulum of the esophagus is successful and might be the method of choice. PMID- 17402214 TI - [Penile cancer]. PMID- 17402215 TI - Exposure to environmental tobacco smoke in pregnancy and lead level in maternal blood at delivery. AB - OBJECTIVES: The particular purpose of our study was to assess the impact of environmental tobacco smoke (ETS) on the individual variability of blood lead levels in pregnant women as earlier publications demonstrated the increased blood lead in smokers. MATERIALS AND METHODS: The material consisted of 240 pregnant women who participated in a prospective cohort study on vulnerability of the fetus and infant to environmental hazards. The enrolment included only non smoking women with singleton pregnancies between the ages of 18-35 years. Whole blood lead concentrations were determined using inductively coupled plasma mass spectrometry. RESULTS: On average, blood-lead measured in pregnant women at delivery was low (GM = 1.7 microg/dL; 95% CI:1.6-1.8 microg/dL) and none of them showed levels above 4.0 microg/dL, but persons reporting exposure to ETS had significantly higher blood lead level (GM = 1.9 microg/dL; 95%CI:1.8-2.1 microg/dL) than those free from this exposure (GM = 1.6 microg/dL; 95%CI:1.5-1.7 microg/dL). In order to single out the effect of the ETS exposure from the confounding variables, we used the stepwise multivariate linear regression for log blood-lead in maternal blood as dependent variable and a set of independent variables, such as age, weight of women before pregnancy and their education level. The results of the analysis showed that all the independent variables included in the model explained 11% of total blood-lead variability among the study women. The strongest component of variance was attributed to ETS exposure (5%), age (3%), education level (2%) and weight (1%). Inclusion into the model of other variables, e.g., residence area and traffic intensity did not improve the proportion of explained variability. CONCLUSIONS: The reason for higher levels of blood-lead in the ETS-exposed women may result from the fact that tobacco smoke contains lead. However, it is possible that inhaled tobacco smoke also increases the absorption of lead from particulate matter deposited in the bronchial tree. PMID- 17402216 TI - Serum Clara cell protein as an indicator of pulmonary impairment in occupational exposure at aluminum foundry. AB - OBJECTIVES: Although some of the exposures in aluminum (Al) smelting have been well characterized, and respiratory disorders in aluminum production workers are well known, the relationship between internal aluminum loads and appropriate lung biomarkers have not been elucidated. The aim of our work was to carry out a comprehensive investigation in workers employed in the Aluminum Foundry Casting Department with special reference to currently existing hygiene standards, known as threshold limit values (TLV) based on aluminum effects on the respiratory system. The measurement of serum anti-inflammatory Clara cell protein (CC16) was employed as a peripheral marker of the lung epithelium function. MATERIALS AND METHODS: A group of 50 casting smelters, 5 locksmiths, 11 sawyers and auxiliary workers exposed to dust containing 14% of aluminum, and a group of 42 controls were examined. Respiratory function tests were performed and forced volume capacity (FVC), forced expiratory volume in 1 s (FEV1), forced expiratory volume in the first percent (FEV1%), forced expiratory flows in 50% VC (FEV50), and markers of foundry workers' exposure and body burden, Al concentration in the breathing zone, blood and urine, biomarkers of the effects of exposure, concentration of CC16 and hyaluronic acid (HA) in serum were determined in all examined workers. Additional measurements comprised determinations of serum iron (Fe) levels, myeloperoxidase (MPO), eosinophil cationic protein (ECP), immunoglobulin E (IgE), glutathione S-transferase (GST), and superoxide dismutase (SOD) activity in erythrocytes. RESULTS: The group of casting smelters was characterized by the highest levels of aluminum in urine (Al-U) (43.7 microg L( 1)), high levels of MPO, ECP and IgE, high SOD activity, low CC16 levels, and low activity of GST. Lower Al-U excretion was observed in locksmiths (35.2 microg L( 1)) and sawyers (21.7 microg L(-1)). Serum CC16 proved to be the most sensitive biomarker, showing high inverse relationship with serum Al (Al-S) concentrations in casting smelters (p = 0.006). CONCLUSIONS: The study showed that in conditions of occupational exposure, dusts containing Al2O3 < 1 mg m(-3) cause changes in the respiratory system and biomarkers in serum, especially in CC16, connected with altered functioning of this system. Changes in concentrations of the examined biomarkers and also in respiratory parameters of the study subjects were observed when Al-U concentration was > 40 microg L (-1). PMID- 17402217 TI - Use of generalized linear mixed models to examine the association between air pollution and health outcomes. AB - BACKGROUND: Time-series and case-crossover are two techniques that are widely used for assessing the short-term impact of ambient air pollution exposure on health. MATERIALS AND METHODS: The generalized linear mixed model (GLMM) methodology is proposed here to study the association between ambient air pollution and health outcomes. Poisson random-effects models are applied to analyze the clustered counts, where the groups of days, determined by the triplet , form the clusters. The proposed technique uses a nested structure for the clusters and allows random-effects for hierarchical factors. A random intercept in the models adjusts for different levels of counts among the clusters. A fixed slope represents a common response to the exposure. RESULTS AND CONCLUSIONS: The obtained results are consistent with those generated by a classical approach (for example the case-crossover technique). The GLMM technique is a valid alternative methodology for studying air health effects. PMID- 17402218 TI - Evaluation of the use of low flow passive sampling technique in offset printing plants. AB - OBJECTIVES: The aim of this study was to evaluate the suitability and applicability of low-flow passive tubes for sampling of organic solvents in occupational environment. MATERIALS AND METHODS: Laboratory and field studies were conducted to continue the evaluation of low-flow diffusive sampling combined with thermal desorption and gas chromatographic/mass spectrometric analysis in occupational hygiene surveys. Passive sampler tubes with Tenax GR adsorbent were employed to assess exposures to organic solvents in 10 small sheet-fed offset printing plants in Finland. The retention of the solvent compounds in the samplers was investigated using laboratory chamber studies. Active sampling with activated carbon tubes served as the control method. RESULTS: The laboratory tests showed that passive tubes had good retention, stability and reproducibility for the solvent compounds. Parallel passive and active sampling in both breathing zone and area monitoring in the printing plants yielded similar measurements though passive sampling tended to give slightly higher concentrations (by 5 to 12%). The suitability of passive sampling to be done by workers was also confirmed. The study also shows that exposure to organic solvents can be effectively reduced by the use of vegetable oil-based cleaning fluids. Especially, since ventilation and other protective measures are often inadequate in small facilities, the use of toxic cleaning solutions should be avoided. CONCLUSIONS: Passive sampling tubes with capillary orifice analyzed with thermal desorption GC-MS are well suited for measuring occupational solvent exposure. Method is well suited for sampling done by workers. PMID- 17402219 TI - Individual susceptibility to noise-induced hearing loss: choosing an optimal method of retrospective classification of workers into noise-susceptible and noise-resistant groups. AB - OBJECTIVES: Individual susceptibility to noise-induced hearing loss (NIHL) depends on the interaction between intrinsic and environmental factors. To proceed with the study on NIHL susceptibility genes an appropriate selection of workers susceptible and resistant to noise is crucial. The aim of the study was to compare four different methods of subject classification by the susceptibility to NIHL in a group of 949 workers of an electric power plant exposed to steady state noise at the workplace. MATERIALS AND METHODS: One method based the classification of the workers on the international reference standard ISO 1999:1990; from the entire group of workers, 10% of the subjects with the worst hearing thresholds (HT) in the model were categorized as susceptible to NIHL, whereas 10% of the subjects with the best HT were categorized as resistant to noise. According to three other methods, the entire group of workers was first divided into subgroups by age, duration of employment and the level of noise, and then 10% of the subjects at each HT extreme were selected. RESULTS: The first classification allowed to achieve an excellent separation between HT of the susceptible and resistant subgroups. The susceptible subgroup was significantly younger than the resistant one, showed a shorter duration of employment and a lower level of noise exposure, which is in line with the definition of increased vulnerability to NIHL. The three other methods produced poorer separation of HT with smaller or no gap between HT values in subgroups. CONCLUSIONS: The selection of subjects from the entire worker population of a given industry based on the ISO 1999:1990 standard can be regarded as the most reliable method of classification of noise-susceptible and noise-resistant workers to be used in the future genetic studies on NIHL susceptibility genes. PMID- 17402220 TI - Self-medication with antibiotics in Lithuania. AB - OBJECTIVES: Excessive and not always proper use of antibiotic give rise to numerous problems, of which antimicrobial resistance, currently cause for worldwide concern, is the major one. Few single studies of antibiotic use have been carried out in some countries. This study was performed to estimate the prevalence of antibiotic use in the general population of Lithuania with special interest in self-medication with antibiotics and sources of their acquisition. MATERIALS AND METHODS: Structured questionnaires on antibiotic use during the last 12 months were mailed to randomly selected adults and 746 of them were finally analyzed. RESULTS: It was found that 39.9% of respondents reported antibiotic use during the last 12 months preceding the study and 53.2% of those used them in self-medication. In general, 22.0% (95%CI: 19.1-25.1) of respondents used antibiotics without prescription, whereas 45.0% (95%CI: 41.3-48.7) of them used antibiotics for intended self-administration. Adjustment for all the factors revealed the impact of the occupation, place of residence and presence of chronic disease on self-medication with antibiotics. Representatives of managerial, executive and professional occupations used non-prescribed antibiotics 8.38 times more often (95% CI: 1.76-39.91, p = 0.01) than retired people. Healthy people showed the tendency to self-medication 2.04 times more frequently than those with chronic diseases (95%CI: 1.11-3.75, p = 0.02). Rural people used non-prescribed antibiotics 1.79 times more often than inhabitants of urban areas (95%CI: 1.00 3.18, p = 0.049). Community pharmacies proved to be the most frequent (86.0%) source of over-the-counter antibiotics. Tonsillitis, bronchitis, and upper respiratory infections were the major reasons for self-medication with antibiotics. CONCLUSIONS: The high prevalence of self-medication with antibiotics was found in Lithuania. The study indicated the need for more strict control of antibiotic sales and promotion of education of the correct use of antibiotic among Lithuanian people. PMID- 17402221 TI - Palpation of the testes in draftees: acceptance on the occasion of muster and implication for health promotion. AB - OBJECTIVES: Testicular cancer is the most prevalent malignant neoplasm in young men. The key problem in screening for malignant testicular tumors and their main risk factors (undescended testes and testes which descended only after medical treatment) is to reach the target group of "young men". MATERIALS AND METHODS: In five different military medical centers of German Armed Forces, 1600 young draftees were investigated by a standardized interview at recruitment examination between 2001 and 2003. RESULTS: Ninety-five percent of respondents reported that they had been palpated at testes during physical examination. The percentage of draftees not palpated at their testes varied between muster centers, but did not depend on the physician's gender. Only two draftees had refused explicitly the palpation of testes. The prevalence of observed pathologic alterations of the testes was 1.69%, including two cases of cryptorchidism, 19 of varicocele, and 6 cases with other benign alterations. CONCLUSIONS: The simple and effective examination of testes should not be restricted to draftees, but promoted by compulsory health insurance funds, offered and performed routinely by all physicians who have access to the target group of "young men". PMID- 17402222 TI - [Procuste cephalometry]. AB - Procrustean cephalometrics employs Procrustes's methods to analyze cranio-facial and orthodontic structures. Using this technique, which is structured on a rigorous mathematical base, orthodontists can avoid the biases and limitations of traditional analyses based on index angles and linear measurements. The starting data are the coordinates of the anatomic landmarks discerned on a profile cephalometric X-ray. Employing this data, orthodontists can calculate patients' type and stage of growth. These factors, representative of the normal variability of each patient, make it possible for the orthodontist to calculate an individualized reference base: a harmonious face in good balance of the same type and at the same growth stage as the patient. Finally, orthodontists make the best possible superimposition of this personalized, "ideal" radiographic sketch on the patient's headplate, by using anatomic landmarks, not a reference plane, to determine the areas that are out of balance and identify the anomalies that will need to be corrected during treatment. PMID- 17402223 TI - [Adaptation of mastication in response to the characteristics of the individual or the food]. AB - This article describes how all individuals formulate an individual masticatory adaptation process to deal with the special characteristics of the food they are chewing. It begins by briefly reviewing the act of mastication and the principal techniques that are used to observe it, focusing especially on electromyography. An overall study of mastication must include an analysis of its result, that is, the bolus of food that it prepares for swallowing. The masticatory act varies with each individual; it has been clearly established that people do not effect the same mandibular movements when they chew. Some people will need more chewing cycles to treat the same bolus of food than others will and the amplitude and duration of muscular contraction will vary from individual to individual. But a certain reproducibility does exist for each subject. The patients' dental condition will, to a large extent, dictate how they chew. The more prosthetic replacements they have in their mouths, the more cycles they will require for mastication. Age, too, plays a role. Older people have diminished perception thresholds and, therefore, take longer to react. If their dentitions are relatively complete and healthy, they simply need to increase the number of chewing cycles for a given portion of food. And, of course, the texture of the foodstuff plays a part; the harder it is the greater the number of cycles will be. PMID- 17402224 TI - [The Class II subdivision or acquired unilaterally dominant mastication syndrome]. AB - Unilateral disto-occlusion of the molars and the canine teeth, described by Angle as Class II subdivision, is often characterized by asymmetric development of the maxillae in conjunction with a disturbance in physiological mastication. In addition to the unilateral malposition of the molars and the canine, this Class II subdivision condition is routinely accompanied by an ensemble of symptoms that the authors propose regrouping under the designation "children's acquired unilaterally dominant mastication syndrome". They use several clinical examples to illustrate the diagnosis of this condition that they consider to be a syndrome of craniofacial asymmetry. PMID- 17402225 TI - [Sagittal and vertical dentoalveolar compensation in hyperdivergent malocclusion: control using the Straight Wire Integrated In Normal Growth (SWING) system]. AB - The "Straight Wire Integrated In Normal Growth" (S.W.I.N.G.) concept invented by Planch65 suggests treating high angle cases by the so-called "anti-clockwise rotation mechanics". The benefits of this method are illustrated by clinical examples covering common malocclusions. PMID- 17402226 TI - [Pathogenic occlusion and functional occlusion: definition of completion]. AB - The masticatory system is widely adaptable and tolerant especially in young people ; this characteristic can be helpful but it can also incur risks because there are limits to everything. Guided solely by the insubstantial support provided by data derived from evidence-based medicine, a researcher would have to be both excessively clever and unusually cunning to define the borderline between physiological and pathological occlusion. Accordingly, the optimization of masticatory function through the intermediary of efficient occlusal interaction of upper and lower teeth remains a fundamental common sense objective of all dental treatment. This common sense is based on the fundamental principles of simplicity and of ergonomy that govern all biological "mechanisms ". By respecting the principles of "centering, stabilization, and guidance", dentists will adhere to a simple but precise manual for preserving or creating, if need be, the good occlusal relationships that encourage unfettered mandibular movement (fitting) and symmetrical global functioning (centring), and absence of interference (guiding). PMID- 17402227 TI - [Dentofacial orthopedics and craniomandibular dysfunction]. AB - There has been a recent recrudescence of cranio-mandibular disorders (CMD), predominantly affecting women. Even children can be affected by CMD, often under diagnoses by their families and by medical professionals. Therefore, a multidisciplinary approach is important when evaluating the variety of diagnostic and etiological problems. The search for postural etiology, whether lingual or dental, is always necessary to better act on the etiology of these disorders. The involvement of orthodontics, in the prevention of CMD, has three impacts: functional, aesthetic and psychological, allowing for a better balance of the cranio-mandibular system. This makes the treatment of cranio-facial orthopedics, a successful choice in the recovery of CMD. PMID- 17402228 TI - Application of guar gum biopolymer in the prescription of tablets with sodium ibuprofen--quality tests and pharmaceutical availability in vitro. AB - The increasing interest of the technology of drug form in natural biopolymers has become the reason for undertaking investigations on the possibility of guar gum application in the prescription of oral solid form of a drug. Alternative compositions and technology of the production of tablets of regulated in time sodium ibuprofen release were worked out for children. Two series of tablets were prepared with guar gum (5 and 10% content) and a series without the biopolymer. The tablet mass in each case contained keryostatic sorbitol and bioadhesive polyvinylpyrrolidone. All tablets were tested as regards the quality of production, compliance with the requirements of Polish Pharmacopoeia VI and potential therapeutic usefulness, manifestation of which is pharmaceutical availability of the therapeutic agent (sodium ibuprofen). The tests demonstrated that the produced tablets with sodium ibuprofen have proper physicochemical properties, in compliance with Polish Pharmacopoeia VI requirements. Application of biopolymer of guar gum type as adjuvant substance contributes to the improvement of the tablet hardness parameters and prevents technological problems (lining mixture of powders to tableting machine punch). The designed tablets demonstrate proper pharmaceutical availability of over 80%. Introduction of guar gum into their prescription prolonged their disintegration time and the rate of sodium ibuprofen release, which predisposes the produced form of a drug to have the function of a tablet with slowed-down release. PMID- 17402229 TI - Micellar solubilization of selected non-steroidal therapeutic agents by new surface-active agents of the class of the products of oxyethylation of ursodeoxycholic acid. AB - A new class of non-ionic surface-active agents were synthesized by means of oxyethylation of ursodeoxycholic acid (UDOCh acid) with the application of modified generation of catalysts in the form of a model prodrug. Basic viscosity values ([eta], Meta, Ro, Robs., Omga) as well as the analytic level of hyrohilic lipophilic balance HLB in the notation of Griffin, Davies and 'HNMR method were determined. In the state of equilibrium the solubilizing properties of aqueous solutions of the products of oxyetyenation of UDOCh acid x nTE were estimated with respect to non-steroidal therapeutic agents such as diclofenac, ibuprofen, ketoprofen, and naproxen. The surface activity of solubilizers of UDOCh acid > or = nTE = 30 type and the thermodynamic stability deltaGm of an adduct emerging in the state of equilibrium were determined. PMID- 17402230 TI - [Results of treatment of orbital fractures with polydioxanone sheet]. AB - Orbital fractures are one of the most frequent consequences following midfacial trauma. If not treated they can lead to serious optical complications as: double vision (diplopia), restriction of ocular motility, eyeball sinking (enophthalmos). Autogenic bone grafts although still wide and effectively used for reconstruction of the fractured orbital walls present some disadvantages. This is morbidity of the donor site, unforeseen resorption, time-consuming procedure and longer postoperative care. Because of that introduction of new materials for orbital reconstruction seems to be desirable. The aim of the work was to evaluate usefulness of the polydioxanone sheets (PDS) as a reconstructive material for orbital floor fractures. MATERIAL AND METHODS: From 1. 09. 2004 to 1. 02. 2006 111 patients with orbital fractures were treated in the Department of Maxillofacial Surgery Klinikum Minden. Age of the patients was between 15- 89 years (mean 43 y.); m:w ratio 78:33. There were 54 isolated orbital floor fractures (in 3 cases with additional medial wall fracture) and 57 zygomatico orbito-maxillary fractures. The diagnosis was based on the clinical picture and coronal CT scans. The patients were operated through a transconjunctival (72%) or an infraorbital-Mustarde (28%) access and the orbital floor (medial orbital wall) was reconstructed with PDS sheet. Control examination was performed immediately, 1 moth and 6 months after the operative treatment. RESULTS: There were 47% true "blow-out", 34% "trap-door" and 19% "en-clapet" fractures. The most important preoperative symptom was double vision (23%), restriction of ocular motility (18%), enophthalmos (3,6%), impairment of function of the infraorbital nerve (41%). Size of the defect was 3+/-1,13 cm(2) in "blow out" fracture, 1,8 +/-0,9 cm(2) in "trap-door" and 2+/-0,5 cm(2) in ,"en-clapet" ones. The primary reconstruction was successful in 97,3% of the cases. Any inflammation or reaction against implant was noted. Persistent double vision was present in 2,7%, restriction of ocular motility in 1,8% and enophthalmos in 0,9% after the primary procedure. These patients were submitted to reoperation. In 1 case badly positioned PDS sheet causing diplopia was removed. In another one, eyeball movement restriction was due to adhesions between the sheet and periorbital tissue. Visual status of this patient has improved after adhesiolysis. In 1 patient with a large bony destruction reconstruction only with PDS sheet was an inadequate treatment. In 7,2% of patients disturbances in the field of innervation of the infraorbital nerve were present 6 months after the surgery. CONCLUSIONS: Alloplastic, resorbable PDS sheets in most cases were a valuable material for the reconstruction of the orbital floor (medial orbital wall). Mechanical properties of PDS seem to be not sufficient for the reconstruction of extremely large bony defects. In these cases use of autogenic bone grafts or a titanium mesh should be rather considered. PMID- 17402231 TI - [Membrane transport of non-homogeneous non-electrolyte solutions: on role of volume flows in creation of concentration boundary layers in binary solutions]. AB - The mathematical model of the thickness of concentration boundary layers controlling by concentration Rayleigh number and volume flows for binary non electrolyte solution was presented. The equations determining of this model can be used to numerical calculations. PMID- 17402232 TI - A development of the generalized Spiegler-Kedem-Katchalsky model equations for interactions of hydrated species in transport through polymeric membranes. AB - A new analysis is presented of the relations between multicomponent friction coefficients and hydration numbers which describe non-ionic and multicomponent solutes transport in membranes in the framework of generalized Spiegler-Kedem Katchalsky frictional model equations. In the framework of this model the expressions linking the permeability parameters of the membrane (Lp, sigma, omegaik) with the friction coefficients (fik) for n-component non-ionic hydrated solutions were presented. This analysis is applied to calculation the frictional and phenomenological coefficients change with hydration numbers of solutes for binary and ternary non-ionic solutions was presented. PMID- 17402233 TI - Practical forms of entropy production for single-membrane system and binary non electrolyte solutions. AB - Linear non-equilibrium thermodynamics (LNET) has been used to express the entropy production in single-membrane system representing the true forces (mechanical and osmotic pressures difference) and flows (volume and solute flows) in a homogeneous or non-homogeneous binary non-electrolyte solution. On the basis of Kedem-Katchalsky model equations of entropy production in single-membrane system in practical forms were described. PMID- 17402234 TI - Effects of copper (I) oxide on growth and biochemical compositions of two marine microalgae. AB - In copper based antifouling (AF) paints Cu (I) oxide was largely used as booster biocide. In this study effect of Cu (I) oxide on two marine microalgae, Tetraselmis suecica and Dunaliella tertiolecta was demonstrated. EC50 (96 hr) concentrations estimated for T. suecica and D. tertiolecta were 1.3 mg l(-1) and 1.34 mg l(-1), respectively. Copper (I) oxide induced changes in growth, chlorophyll, carbohydrate and protein contents were observed in T. suecica and D. tertiolecta. At low concentration of 0.0625 mg l(-1), 3-26% and 1-16% growth stimulation was observed in T. suecica and T. tertiolecta respectively. Increasing Cu (I) oxide concentrations proportionately decreased the carbohydrate and protein contents. This study clearly indicates the toxicity of excessive Cu (I) oxide on growth and biochemical compositions of T. suecica and D. tertiolecta. PMID- 17402235 TI - Environmental impact assessment for alternative-energy power plants in Mexico. AB - Ten Environmental Impact Assessment Reports (EIAR) were reviewed for projects involving alternative power plants in Mexico developed during the last twelve years. Our analysis focused on the methods used to assess the impacts produced by hydroelectric and geothermal power projects. These methods used to assess impacts in EIARs ranged from the most simple, descriptive criteria, to quantitative models. These methods are not concordant with the level of the EIAR required by the environmental authority or even, with the kind of project developed. It is concluded that there is no correlation between the tools used to assess impacts and the assigned type of the EIAR. Because the methods to assess impacts produced by these power projects have not changed during 2000 years, we propose a quantitative method, based on ecological criteria and tools, to assess the impacts produced by hydroelectric and geothermal plants, according to the specific characteristics of the project. The proposed method is supported by environmental norms, and can assist environmental authorities in assigning the correct level and tools to be applied to hydroelectric and geothermal projects. The proposed method can be adapted to other production activities in Mexico and to other countries. PMID- 17402236 TI - Concentration of heavy metals in the food web of Lake Egirdir, Turkey. AB - The concentration of heavy metals (Cr, Cd, Hg and Pb,) was determined by atomic absorption spectrophotometry in sediment, water, plankton and fish samples from Lake Egirdir in South-Western Turkey. Lake Egirdir is an important bird nesting and visiting areas, and it is also used as irrigation and drinking water source. The heavy metal concentrations were in the order Cd > Pb > Cr > Hg in water, Cr > Pb > Cd > Hg in sediment, Pb > Cd > Cr > Hg in plankton, Cd > Cr > Pb > Hg in the muscle and Cd > Pb > Cr > Hg in the gills of Ctenophatyngodon idella. The significant differences of concentrations occurred in water, sediment, plankton and fish tissues (Chi square test, p < 0.05). The concentrations of the heavy metals in water and fish tissues of three years old, were above the maximum permissible level for human consumption. PMID- 17402237 TI - Relative toxicity of spent lubricant oil and detergent against benthic macro invertebrates of a west African estuarine lagoon. AB - The relative acute toxicity of spent lubricant oil and detergent was evaluated against hermit crab, Clibanarius africanus (Aurivillus) and periwinkle, Tympanotonus fuscatus (L) from the Lagos lagoon in laboratory bioassays. Based on the derived toxicity indices, the detergent (96 hr LC50 = 5.77ml/l) was found to be 1.73 times more toxic than spent engine oil (96 hr LC50 = 10.01 ml/l) when acting singly against C africanus and 18.73 times (96 hr LC50-48.67 ml/l) more toxic (96 hr LC50 = 911.57 ml/l) when acting singly against T. fuscatus. On the basis of the computed susceptibility factors, C. africanus was found to be about eight times and ninety-one times more susceptible to the toxic effect of detergent and spent lubricant oil respectively. The randomized analysis of variance (ANOVA) showed that there was significant difference (Fcal 58.83 Ftab 3.87; DF 13; p > 0.05) between all treatments of spent lubricant oil and detergent during the 96 hr exposure period of test animals. At 5% level of significance the Student Neuman-Keuls (SNK) test further revealed significant differences in the mean mortality response of test animals exposed to toxicants at all concentrations and untreated control. The results obtained in this study suggest that the estuarine benthic macroinvertebrates, which play key roles in the environment, may serve as useful in-situ sentinels for biomonitoring studies of petroleum pollutants in fragile aquatic ecosystems such as the Lagos lagoon. PMID- 17402238 TI - Effects of vesicular-arbuscular mycorrhizal (VAM) fungi on the seedling growth of three Pistacia species. AB - The experiment was undertaken to test the efficiency of inoculation of vesicular arbuscular mycorrhizal (VAM) fungi on the seedling growth of three Pistacia species used as rootstocks. The stratified Pistacia seeds were inoculated with VAM fungi. The highest rate of inoculated roots was 96.7% in P. khinjuck seedlings with G. clarum and G. etunicatum, 83.3% in P. vera seedlings with G. caledonium and 73.3% in P. terebinthus seedlings with G. caledonium. Mycorrhizal inoculations improved seedling height only in P. terebinthus. Certain mycorrhizal inoculations increased the leaf N, but not P and K contents. Seedlings inoculated with G. caledonium had higher reducing sugar contents. It was concluded that pre inoculated Pistacia seedlings could have a better growth in the harsh field conditions. PMID- 17402239 TI - Morphometric and electrophoretic analysis of 13 populations of Anatolian black pine in Turkey. AB - The genetic variation in populations of Anatolian black pine (Pinus nigra Arn. subsp. pallasiana (L.) Holmboe.), one of the species covering large areas in Turkey, was investigated. Open pollinated seeds were collected from 13 populations in a natural distribution range. Six characters of seeds (length, width, ratio of length to width, weight/1000 seeds) and seedling characters (cotyledon number and hypocotyls height) and two enzyme systems viz. leucine aminopeptidase (LAP) and glutamate oxaloacetate transaminase, (GOT) were investigated. Significant differences were detected among the populations for the morphological characters. In addition, isozyme patterns of two enzyme systems revealed that LAP has two loci (one with 2 alleles and the other with 3), while GOT has three loci (two with 3 alleles and the third one with 2 alleles). Polymorphic loci were 74% on the average. The mean number of alleles per loci was 1.94 and expected heterozygosity was 19%. The mean total genetic diversity was calculated as 0.203; the mean gene diversity within populations was determined as 0.188, and the average between subpopulations diversity was 0.016. The relative magnitude of genetic differentiation among subpopulations was measured as 0.074 indicating that only 7.4% of the total genetic diversity was there between populations. Average genetic distance was 0.093 according to Gregorius. Nei's genetic distance was 0.022. PMID- 17402240 TI - Growth variation of Paulownia Sieb. and Zucc. species and origins at the nursery stage in Kastamonu-Turkey. AB - The present study was carried out on Paulownia tomentosa (6), Paulownia elongate (4), Paulownia fortunei (5) and Paulownia fortunei x tomentosa (1) origins at Kastamonu Forest Nursery. The seedling height growth (SH), root collar diameter (RCD), dry root percentage (DRP) and seedling percentage (SP) of one-year old seedlings of the origins were studied for comparing growth performance of the species and origins and discussing for guidance to field stage based on nursery stage. As a result, P. tomentosa x fortunei hybrid has the highest SH (72.62 cm) among the used species. Significant difference was determined among the origins as to the SH and DRP. The maximum SH were determined for P. tomentosa Beijing Daxin (81.32 cm) and P. elongata Beijing-Daxin (80.76 cm) origins. The maximum DRP were determined for P. tomentosa Anhui-Tongling (77%) origin. There were no significant differences based on RCD and SP among the species and the origins. Among the parents, there were important diversities for SH and RCD. As a final remark, observations and evaluations of the Paulownia studies should be included with the clonal variation for further studies because of the observed growth variations within the population. PMID- 17402241 TI - Effect of complete and skeleton photoperiods in baya weaver, Ploceus philippinus. AB - A study was performed to investigate the effects of complete and skeleton photoperiods on gonadal stimulation in baya weaver, Ploceus philippinus. In August 2003, five groups of acclimatized birds (n = 7 each) were exposed to 6L: 13D: 1L: 4D, 6L: 6D: 1L: 11D, 11L: 13D and 13L: 11D. Birds were refractory and their exposure to 6L:13D:1L:4D, 6L:6D:1L:11D, 11L:13D and 13L:11D did not evoke any response. This clearly indicates that the birds were insensitive to the stimulatory effects of these photoperiods and perceived them as short days. An attempt was made to test the responsivity by subjecting all these groups to long day exposure after 16 weeks of experiment. Under 16L: 8D, birds did not show a response during the first 4 weeks of exposure (week 16 to week 20), but thereafter there was a significant photoinduction except 11L:13D (week 24), but the rate and magnitude was different. Until the end of the experiment (32 weeks), all groups of birds had maximum testicular volume except 11L:13D. Birds pre exposed to 6L:13D:1L:4D, 6L:6D:1L:11D, 11L:13D and 13L:11D responded to 16L:8D, which suggests that these photoperiods helped the birds in recovering their photosensitivity. The results from this experiment indicate that baya weaver interprets a light period depending when the light falls relative to its endogenous circadian system. PMID- 17402242 TI - Impact of endosulfan on the profiles of phospholipids at sublethal concentration in the male Heteropneustes fossilis (Bloch). AB - Male Heteropneustes fossilis were exposed for 30 days at sublethal concentration (0.002 ppm) during different phases of its annual reproductive cycle. Its impact on total phospholipids (TP), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatdylinositol (PI) and phosphatidylethanolamine (PE) were measured in liver, plasma and testes. During preparatory phase, in general, the levels for TP, PC, PS, PI and PE decreased after endosulfan exposure in the above tissues. During prespawning and spawning phases, the phospholipids also showed decreasing trend. The postspawning phase, exhibited decline in hepatic levels of PS and PI only and remained unaffected in the other two tissues. During the resting phase too, the hepatic levels of TP, PS and PI declined and remained unaltered in others. The present results indicate that endosulfan have very selective effects on phospholipids classes during different phases of the annual reproductive cycle interfering with the production of lipid deprived energy i.e. vitellogenin. In general, endosulfan has inhibitory role during reproductive growth affecting phospholipid biosynthesis via hepatic enzyme systems as well as by hormonal imbalance. PMID- 17402243 TI - Distribution and diversity of ground beetles in Baskonus Mountain National Park of Turkey. AB - This study was carried out in National Park Baskonus Mountain (Kahramanmaras, Turkey), in Mediterranean region from April to October of 2004 and 2005. A total 31 species of ground beetles (Coleoptera, Carabidae) were recorded in the pitfall traps. Among them, Carabus (Archicarabus) gotschi caramanus Fairmaire, 1886, Carabus (Procrustes) coriaceus mopsucrenae Peyron, 1858 and Nebria (Nebria) hemprichi Klug, 1832 were the most abundant species. Distribution, diversity and monthly distribution of species in steppes, forest edge and forest interior, and chorotypes and ecology of these species were separately given in this study. It was recorded that the diversity of ground beetles was significantly higher in the forest edge and steppes than in the forest interior. There was no significant difference in the diversity of ground beetles in the steppes and the forest edge. PMID- 17402244 TI - The effects of walnut and pine leaves on bread wheat growth and frequence of common weed species in the East-Mediterranean region. AB - A field experiment was carried out to determine the effects of walnut (Juglans regia L.) and pine (Pinus sp.) leaves on bread wheat (Triticum aestivum L.) growth and weed control, during 2001-02 and 2002-03 growing seasons in East Mediterranean region of Turkey. In this research, the treatments were: applications of ground walnut leaves (GWL) and ground pine leaves (GPL) at 19 g/m2 dose at Zadoks-11, applications of 200, 400 and 800 cc/m2 of walnut leaves extract (WLE2, WLE4 and WLE8 respectively), pine leaves extract (PLE2, PLE4 and PLE8 respectively), mixture extract of 200 cc walnut and 200 cc pine (ME), herbicide application (HA) at 8 g/m2 dose at Zadoks-21, and control (C) without any treatment. The results have indicated that the effects of treatments were significant for grain yield (GY), plant height (PH), flag leaf length (FLL), weed number per square meter (WN), but non significant for grain number per head (GN), grain weight per head (GW), 1000 grain weight (1000 GW), flag leaf width (FLW), head number per square meter (HN), vegetative period (VP), grain filling period (GFP) and days to maturity (DM). PMID- 17402245 TI - The assessment of environmentally sensitive forest road construction in Calabrian pine forest areas of Turkey. AB - Forest road construction by bulldozers in Calabrian Pine (Pinus brutia Ten.) forests on mountainous terrain of Turkey causes considerable damage to the environment and the forest standing alongside the road. This situation obliges a study of environmentally sound road construction in Turkey. This study was carried out in 4 sample sites of Antalya Forest Directorate in steep (34-50% gradient) and very steep terrain (51-70% gradient) conditions with bulldozer and excavator machine and direct damages to forest during road construction was determined, including forest area losses and damages to downhill trees in mountainous areas. It was determined that in steep terrain when excavators were used, less forest area (22.16%) was destroyed compared to bulldozers and 26.54% less area in very steep terrain. The proportion of damage on trees where bulldozer worked was nearly twofold higher than excavator was used. The results of this research show that the environmentally sensitive techniques applied for the road construction projects are considerably superior to the traditional use of bulldozers on steep slopes. The environmentally sound forest road construction by use of excavator must be considered an appropriate and reliable solution for mountainous terrain where areas of sensitive forest ecosystems are to be opened up. PMID- 17402246 TI - Some properties of new nesting areas of sea turtles in north-eastern Mediterranean situated on the extension of the Samandag Beach, Turkey. AB - One of the most endangered species in tropical seas, Chelonia mydas (green turtles) prefer undisturbed sand beaches to lay their eggs. While Caretta caretta lays their eggs all over the Mediterranean beaches of Turkey, C. mydas nesting locations are limited with north-eastern site (Alata, Kazanli, Akyatan and Samandag) of the country and some beaches in Cyprus. Until 2003, no nesting place of both species between Akyatan and Samandag was recorded. From north to south, the beaches examined in 2003 summer are Arsuz, Konacik, Kale, Tr-H-3, Tr-H-2, and Tr-H-1 in addition to well-known and studied nesting beaches of sea turtle species in Hatay Province: Cevlik, Seyhhizir and Meydan. Since the Hatay Provincial Coast between Cevlik and Arsuz is hilly and no stabilized road is available, these small beaches were not known to have both C. mydas and C. caretta nests. Some physical and chemical sand properties, the number of nests belong to both species and selected nest specifications were investigated in this study. Kale Beach was found to be the most important nesting place in this less populated area. Based on results, Kale, Tr-H-3 and Tr-H-1 beaches were recorded to have high hatchling success. PMID- 17402247 TI - Flow of chemical energy in Alwar jheel of Yamuna basin near Allahabad. AB - The water quality, rate of energy transformation, chemical composition of producers and flow of chemical energy were studied in both feeding river Yamuna and Alwar jheel near Allahabad. As the river Yamuna had high value of alkalinity (210.0 mgl(-1)), conductance (518.0 micromhos), dissolved solids (260.0 mgl(-1)), hardness (162.0 mgl(-1)) and chloride (54.6 mgl(-1)) jheel also showed high values of these parameters. The rate of energy transformation from kinetic radiant energy to chemical energy was very high in the jheel 32,315 Cal m(-2) day(-1) of which 25,620 Cal m(-2) day(-1) was contributed by aquatic plants. Out of 11,764 x 10(4) Kcal ha(-1) yr(-1) total energy fixed in the system, producers stored 7,154 x 10(4) Kcal ha(-1) yr(-1) and the rest was lost as heat of respiration. The pattern of storage of energy was different in two groups of producers and thus most of the energy fixed by phytoplankton was stored as protein (56.2%) and less as carbohydrate (11.7%) while aquatic plants stored more energy as carbohydrate (40.8%) than protein (23.2%). The chemical energy obtained from the system was 1,85,000 Kcal ha(-1) yr(-1) and thus only 0.260% of the chemical energy stored by producers was harvested. The potential chemical energy resource in the jheel was 81.4 x 10(4) Kcal ha(-1) yr(-1) of which only 22.6% was harvested in the jheel and there is enough scope for further enhancement. PMID- 17402248 TI - Identification of a sensitive index during fish bioassay of an azo dye methyl red (untreated and treated). AB - Acute and chronic toxicity of methyl red (untreated) was examined on a freshwater fish Poecilia reticulata, using indices viz; mortality, reduction in RBC counts and their morphological abnormality (poikilocytosis and anisocytosis). Similar studies (acute toxicity) were also made in physicochemically and biologically treated methyl red. Data comparison of these four indices revealed poikilocytosis as the most sensitive index, since it measured higher toxicity of methyl red when fish mortality was either minimum at its low concentration (5 ppm) during both acute and chronic toxicity or even nil in the biologically treated 100 ppm methyl red, during acute toxicity. Mortality was next to poikilocytosis though it ranked 1st at higher concentration of methyl red during acute toxicity. The reduction in RBC counts however, was found to be the most sensitive parameter only in case of prolonged exposure (4 weeks) to 5 ppm methyl red. Amongst the four indices used for quantifying toxicity; anisocytosis was found to be the least expressive. Based on these findings we recommend quantification of data on fish mortality and poikilocytosis during acute toxicity whereas reduction in RBC counts and poikilocytosis during chronic exposure to methyl red. PMID- 17402249 TI - Metabolic variation in rice cultivars of contrasting salt tolerance and its improvement by zinc in sodic soil. AB - The severity of Zn deficiency increased with increase in soil exchangeable sodium percentage (ESP) with salt sensitive variety M1-48 scoring 6 at ESP 62 as against only score 3 by salt tolerant variety Pokkali under similar soil conditions. Strikingly, zinc contents were much higher in salt tolerant variety than in salt sensitive one. Zinc application increased zinc concentration in the roots by a factor of 2.85 to 3.87 in Pokkali whereas it rose from 2.37 to 4.35 times in M1 48 depending upon ESP but in the leaves it registered increase of 1.5 to 1.8 times only. In general, the concentrations of reducing sugar were less (about 2.2%) than that of non-reducing (about 3.8%) in both the varieties under normal soil conditions. However, the concentration of reducing sugar doubled (4.2-4.4%) at the highest ESP 62, whereas the concentration of non-reducing sugar though increased (4.1 to 5.1%) but not as vigorously as reducing one. Zinc application reduced the concentration of reducing sugar but not that of non-reducing at similar ESP values. In Pokkali, the concentrations of total sugar increased from 6% at ESP 20 to 9.34% at ESP 62, whereas it registered enhancements of 5.98 to 8.6% in M1-48 under similar conditions. The nitrate reductase (NR) activity decreased with increase in soil sodicity however, the varietal differences in NR activity were wider under Zn-stress than under conditions of applied zinc with Pokkali registering higher NR activities. Carbonic anhydrase activities were higher in salt tolerant variety. Inhibition in carbonic anhydrase activity amounted to 23 and 45% in salt-sensitive variely M1-48 whereas only 19 and 33% in salt-tolerant variety Pokkali at ESP 41 and 62, respectively. The effects of zinc application at higher soil sodicity were more obvious in salt-sensitive variety than in salt-tolerant one. The findings suggest that the tolerance to Zn stress runs parallel to salt tolerance abilities of rice varieties. PMID- 17402250 TI - Water quality and phytoplankton characteristics in the Palk Bay, southeast coast of India. AB - The present study was carried out to determine the water quality in terms of physicochemical characteristics and plankton distribution in the coastal waters of Kattumavadi, Palk Bay for a period of one year from April 2002 to March 2003. Air and surface water temperatures varied from 28 degrees C to 32.50 degrees C and from 27.5 to 32.0 degrees C while light extinction coefficient (LEC) varied between 0.95 and 1.85. Salinity ranged from 26.0 to 34.5 per thousand and the pH ranged between 7.95 and 8.35. Variation in dissolved oxygen content was from 4.15 to 7.18 ml(-1), and the particulate organic carbon (POC) content varied between 0.49 and 2.28 mgCl(-1). Concentrations of nutrients viz. nitrate (2.15 to 8.28 microM), nitrite (0.12 to 0.62 microM), inorganic phosphate (1.28 to 2.15 microM) and reactive silicate (5.15 to 12.52 microM) also varied independently. Chlorophyll a content ranged from 0.28 to 1.48 mg m(-3) and the primary productivity, from 4.19 to 16.08 mgCm(-3) hr(-1). The present study recorded a total of 43 species of planktonic diatoms and two species of blue-green algae. Population density of phytoplankton varied from 18,000 to 34,000 cells l(-1). Percentage composition, of the diatoms showed minimum values during the monsoon season and the maximum values during the premonsoon season. PMID- 17402251 TI - Changes in the level of transaminases in Indian major carp, Labeo rohita exposed to sublethal concentration of tannery and distillery effluents. AB - The activity of alanine aminotransferase (ALAT) and aspartate aminotransferase (AAT) of different tissues of fingerlings of Labeo rohita under the influence of two effluents has been studied. The alanine aminotransferase activity was increased over the control in different exposed periods of tannery and distillery effluent treatments. The alanine aminotransferase in the liver showed increased activity at different periods than that of the muscle, kidney, gill and brain (p < 0.001) (60.09%) over the control during the 40 days exposure in both the effluents treatments. The increased activity of alanine aminotransferase was highly significant (p > 0.001) in all the tissue in tannery and distillery effluents treatments. Similarly aspartate aminotransferase activity was increased over the control in all the treated tissues from 10 to 40 days exposure. But this increase, was not significant in the muscle tissue in distillery and tannery treatements after 10 days exposure. From 10 to 40 days, the activity was increased but a maxmum elevation was observed during 40 days, where the elevation was more in the liver, which was followed by muscle, kidney, gill, brain (brain < gill < kidney < muscle < liver). PMID- 17402252 TI - Allelopathic effect of four weed species extracts on germination, growth and protein in different varieties of Glycine max (L.) Merrill. AB - Allelopathic effect of Ageratum conyzoides L., Cynodon dactylon (L.) Pers., Parthenium hysterophorus L., and Solanum nigrum L. were examined on seed germination, seedling growth, total protein content and protein profile on Ankur, Bhatt, Bragg, PK -416, PS-1042 and Shilajeet varieties of soybean (Glycine max (L) Merill). Aqueous extracts of weeds (10% w/v) showed both inhibitory and stimulatory influence on percent seed germination and seedling growth in different varieties of soybean. On the basis of per cent reduction in different parameters, the variety Bragg and PS-1042, and Shilajeet were resistant and susceptible respectively to different weed extracts, and among weed extracts, S. nigrum was most effective followed by P. hysterophorus compared to others. The total protein content (mg/g f wt.) in different varieties was increased with all the weed extracts except Bragg with C. dactylon and P. hysterophorus, PS-1042 with A. conyzoides and Shilajeet with C. dactylon, in which it was decreased. The protein banding pattern in different varieties not only differ between control and treatments but also among treatments. The order of susceptibility of different varieties with different weed extracts followed the order: Ankur > PK 416 > Bhatt > Shilajeet > Bragg and > PS-1042. PMID- 17402253 TI - Studies on the role of six enzymes in the metabolism of kinetin in mustard aphid, Lipaphis erysimi (Kalt.). AB - The activity of catalase, glutathione peroxidase, superoxide dismutase, O demethylase, ATPase and succinate dehydrogenase, belonging to two main classes of detoxification enzymes (i.e. hydrolases and oxido-reductases), mostly involved in metabolism and degradation of xenobiotics in insects, were assessed under the influence of kinetin, a plant growth regulator (PGR). The nymphs (48-52 hr old) of Lipaphis erysimi (Kalt.) were permitted to feed on radish plant, Raphanus sativus L. treated with kinetin (400 ppm) for 13, 25 and 37 hr. It was found that the activity of catalase, glutathione peroxidase and superoxide dismutase increased significantly when compared with the control of the same age group, which indicated that these enzymes might be playing a significant role in the metabolism of kinetin in this insect. The activity of O-demethylase showed an increase up to 25 hr of the treatment but it decreased under prolonged treatment whereas the activity of succinate dehydrogenase fluctuated insignificantly. ATPase showed a decrease in the activity with the treatment suggesting kinetin's interference in synthesis of ATPase. PMID- 17402254 TI - Acute toxicity of copper sulphate to fresh water prawns. AB - Fresh water prawns, Macrobrachium lamarrei and Macrobrachium dayanum (Crustacea Decapoda) were subjected to static bioassay tests to ascertain the LC50 values of copper sulphate. The 24, 48, 72 and 96 hr LC50 values of copper sulphate for M. lamarrei were 0.38, 0.361, 0.343 and 0.300 mg/l and for M. dayanum were 1.634, 0.988, 0.532 and 0.418 mg/l respectively. Behavioral responses and LC50 values indicate that M. lamarrei were more sensitive to copper sulphate than M. dayanum. PMID- 17402255 TI - Population ecology of a freshwater turtle Kachuga tentoria near Panchnada (Etawah: U.P.) and its role as water purifier. AB - The present study deals with biology, ecology and population dynamics of freshwater turtle Kachuga tentoria and its role as water purifier. The study area Panchnada is the site, where five important national rivers meet together and is preserving an appreciable population of nine species of fresh water turtles. Kachuga tentoria was located at all the sampling stations surveyed by the authors, and hence selected for the present study. Different activities (nesting, incubation, predation and other reproductive aspects), climatic conditions, habitat, population density and morphometric features were worked out in detail. A time bound conservation strategy is needed to save this species from extinction. In situ conservation will be more helpful for the recruitment of the population of this species. It will help in the "hatch and release programme" to clean different polluted national rivers. PMID- 17402256 TI - Accumulation of lead, cadmium and chromium in some plants cultivated along the bank of river Ribila at Odo-nla area of Ikorodu, Lagos state, Nigeria. AB - Heavy metal in soil samples and in washed and unwashed samples of Telfaria occidentalis (ugwu) and Talinum triangulare (waterleaf) cultivated on the bank of river Ribila in Odo-nla village were determined. The soil was moderately polluted with cadmium when compared with Federal Environmental Protection Agency standards. The difference between the unwashed and washed plant samples revealed that metal pollutants exist as superficial contaminants on the foliage surface which is the edible portion and if the foliage portion is washed thoroughly it may be safe for dietary consumption. There is no doubt that continuous discharge of effluent and gaseous emissions from the industries located in this area and dumping of domestic wastes into the river may lead to higher concentrations of these heavy metals in the soil and in the tissue of the leafy vegetables cultivated on the river bank over time. This can eventually lead to pollution of the soil and the cultivated plants, which are ready source of food for the people and other organisms in the food chain. PMID- 17402257 TI - Effects of planting date and time of nitrogen application on yield and sugar content of sweet sorghum. AB - This experiment was conducted at Azad University experiment station to evaluate the effects of planting date and time of nitrogen application on yield and sugar content of sweet sorghum in 2003. Four planting dates (May 4, March 19, June 3 and June 18) and three stages of nitrogen application (3-5/8 leaf stage, boot stage and soft dough stage) were assigned to the main and subplots, respectively. Plant height and diameter, fresh stalk yield, total dry weight, brix value (soluble solids), sugar content and grain yield were affected by planting date. For all the characteristics, the maximum value was obtained by the first planting date. Only stem height, diameter and total plant dry weights were affected by time of nitrogen application. In these cases, nitrogen application at (3-5/8) leaf stage was superior to others. According to the results, sweet sorghum should be planted on the beginning of May and nitrogen be applied at (3-5/8) leaf stage. PMID- 17402258 TI - Body mass and cuticular transpiration of the pulmonate slug Milax rusticus treated with spinach homogenate filtrate. AB - The effects of four concentrations (25, 50, 75, 100%) of spinach homogenate filtrate over a period of 96 hours on the body mass and transpiration rates of the pulmonate slug Milax rusticus, a crop pest found in Benghazi, Libya, are reported. A decrease in body mass and transpiration rate in filtrate treated slug over time was evident and the differences in these factors between control and treated slug were significant (p < 0.01). Tendencies for regaining lost weight and increase in transpiration were observed in the filtrate treated slugs during their seven days of recovery period in normal conditions. Application of spinach homogenate filtrate as a possible contact slugicide of M. rusticus is suggested. PMID- 17402259 TI - Assessment of noise level in Burdwan town, West Bengal. AB - Cities and towns of the world are now facing enormous rise of noise pollution problem due to very high population rise, transport congestion and associated commercial and industrial activities. Burdwan, a district headquarter (100 km away from Kolkata) is one such town where noise pollution is very frequent. In order to assess noise level, noise data were collected from various places of the town by sound level meter with a duration of 30 minutes/location during specified time like 6.00 am, 10.00 am, 1.00 pm, 4.00 pm and 6.00 pm. Most of the monitoring places either belongs to silence category or commercial category areas. From the tabulated data, it was found that sound level lies within the range of 64-85 dB or above in different time at different places. The locations that belong to the silence zone have the noise level up to 90 dB. Statistically noise level in all these zones differ significantly at their peak hours. Noise pollution adversely affects our environment as well as human beings. Sound causes both pathological and psychological disorders in human beings. Implementation of rules and regulations under section 20, 21J, 41, 68(I), 70, 90, 111A of Environment Protection Act, 1986 and of course various technological methods and public awareness are very essential to check noise pollution in Burdwan town. PMID- 17402260 TI - On the occurrence of a double parasitism (copepod and isopod) on the anchovy fish in India. AB - A double parasitism (isopod and copepod) on anchovy fish, Stolephorus commersonii Lacepede, 1803 was recorded from Parangipettai (India) coastal waters during June, 2004. This is the first report from this region and the infestation is discussed in relation to environmental parameters in this short communication. PMID- 17402261 TI - Quo vadis (whither goest thou)? PMID- 17402262 TI - Accessing modern contraceptive technologies in India. AB - The 'unmet' need for reproductive health care in India requires a wide demand net. Besides 'supply' of oral contraceptives, condoms and IUDs, the 'non-supply' approach for increasing contraceptive prevalence is recommended through education about the variety of safe contraceptives and awareness in the use of traditional methods. Expansion of education is partly responsible for the recent decline in fertility in India, however, gender inequality still prevails with poor sex ratio in urban regions, and deficit of around 35 million girls and women from higher mortality compared to males till the age of 30. The community can become a key force in knowledge sharing for greater contraceptive prevalence and also to support its service sector. Practice of.women's reproductive health involves complex socio-cultural-biological interaction in accessing modern contraceptive technologies. A number of interactive factors have been suggested to operate at different hierarchical levels to provide homeodynamic stability to the central event of reproductive health care under a central controller action, the government. While guidelines of the World Health Organization for quality care in family planning should be implemented in rural and urban sectors, efforts should also be made to translate recent advances in contraceptive technology from laboratory to service sector for improving women's reproductive health. PMID- 17402263 TI - Functional status of auditory pathways in hypothyroidism: evoked potential study. AB - Hypothyroidism is known to be associated with mental retardation, motor dysfunction, memory deficits and hearing impairment. In the present study, the functional integrity of the thalamocortical projections to the primary auditory cortex and association cortex has been assessed by using Auditory Evoked Responses i.e Auditory Brainstem (ABR), Mid Latency Response (MLR) and Slow Vertex Response (SVR). Thirty newly diagnosed hypothyroid patients and thirty healthy controls were taken for the study and ABR, MLR and SVR were recorded on computerized evoked potential recorder using 10-20 system of electrode placement. The second recordings for the hypothyroid patients were done 3 months after treatment with attainment of euthyroid states. The present study revealed a slight increase in absolute latency of wave III of ABR in hypothyroid patients and significant decrease in absolute latency of wave III and interpeak latency of I-III after treatment. There was a significant decrease in amplitude of wave V in hypothyroid patients and significant increase in amplitudes of ABR wave I and wave V after treatment. There was a significant increase in latency of wave Na of MLR and wave P2 of SVR in hypothyroid patients. The latencies of waves Na, Pa, Nb of MLR and waves PI and N2 of SVR showed significant improvement with thyroid hormone treatment. The results of the present study indicates that in hypothyroid state there might be slow conduction at the periphery and with treatment there is better recruitment of neuronal pool of the generators of the waves of ABR in the brainstem. We can also conclude that the thalamocortical projections of the auditory pathways are adversely affected in the hypothyroid state and this improves after treatment. PMID- 17402264 TI - Lipid peroxidation and antioxidant status in patients with carcinoma of prostate. AB - Prostate cancer is the most prevalent cancer found in men above the age of fifty years and is frequently diagnosed in men between 45 and 89 years of age with a median age of 72 years. This work was undertaken to assess oxidative stress and anti oxidant status in patients with carcinoma of prostate. Glutathione (GSH), Malondialdehyde (MDA), Super Oxide Dismutase (SOD) levels in Erythrocytes and plasma Glutathione-S-Transferase (GST) levels were estimated in patients with carcinoma of prostate and compared to controls. It was observed that Erythrocyte GSH levels were significantly lower and Erythrocyte MDA & SOD levels were significantly higher in patients with carcinoma of prostate compared to controls. No significant change was observed in case of GST compared to controls. Oxidative stress may be involved in prostate cancer as evidenced by the higher MDA levels and lower GSH levels. The increased activity of antioxidant enzyme may be a compensatory regulation in response to oxidative stress. PMID- 17402265 TI - In vitro action of leukemia inhibitory factor (LIF) on mid-secretory stage endometrial stromal cells collected from hormone-simulated, ovariectomized monkey and maintained in three-dimensional primary culture. AB - Leukemia inhibitory factor (LIF) is a pleiotropic cytokine that is known to play an important role in blastocyst implantation. The putative action of LIF in the regulation of uterine function has been examined using mid-secretory stage monkey endometrial stromal cells cultured on rat-tail collagen type I and treated with recombinant human LIF (rhLIF) or immunoneutralized LIF (in LIF) under serum-free condition. Long-term ovariectomized rhesus monkeys (n=8) underwent simulation of their menstrual cycles with steroid hormones and endometrial tissue samples were collected on cycle day 18; stromal cells were isolated and grown in primary culture on three-dimensional collagen matrix. Significant decline in cellular protein synthesis (P < 0.01) and cell proliferation index (P < 0.05) was observed in cells with increasing doses (0-1000 ng/ml) of rhLIF under serum-free in vitro condition. JAK1 expression in cultured cells increased (P < 0.01) in response to rhLIF as revealed from Western blot and confocal laser scanning microscopic examination, STAT1 and STAT2 expressions were unchanged, while pSTAT3 expression increased (P < 0.01) with increased concentration of rhLIF in culture medium. Autophosphorylation of JAK1 in endometrial stromal cells showed no change with increasing concentration (0.01 to 100 ng/ml) of rhLIF in vitro, but significant (P < 0.05) increase was observed with the time of exposure to rhLIF. Immunoneutralization of LIF or no addition of rhLIF to cultured cells led to significant (P < 0.01) increase in stromal cell proliferation index and significant (P < 0.01) decrease in the level of JAK1 and its autophosphorylation as compared to cells exposed to rhLIF alone. From the present set of experiments we conclude that rhLIF affects the physiological behaviour of monkey mid secretory stage endometrial stromal cells in vitro via the JAK-STAT signaling pathway. PMID- 17402266 TI - Assessment of computer game as a psychological stressor. AB - To simulate the effects of acute psychological stress, the effects of stressful computer game in young adult subjects were assessed by various physiological, psychological and biochemical parameters. The results showed a significant increase in the physiological and psychological markers of stress. It is concluded from these results that computer game can be used as an acute laboratory psychological stressor for future studies on physiological effects of stress. PMID- 17402267 TI - Effect of Sahaj Yoga on neuro-cognitive functions in patients suffering from major depression. AB - Cognitive functions are impaired in Major Depression. Studies on the effects of Yoga on cognitive functions have shown improvement in memory, vigilance and anxiety levels. 30 patients suffering from Major depression (age 18 to 45 years) were randomly divided into two groups: Group 1: (10 males and 5 Females) Patients who practised Sahaj Yoga meditation and also received conventional anti depressant medication. Group 2: (9 males and 6 Females) Patients who only received conventional antidepressant medication. Group 1 patients were administered Sahaj Yoga practice for 8 weeks. Neuro-cognitive test battery consisting of Letter cancellation test (LCT), Trail making test 'A' (TTA), Trail making test 'B' (TTB), Ruff figural fluency test (RFFT), Forward digit span (FDS) & Reverse digit span test (RDS) was used to assess following cognitive domains: Attention span, visuo-motor speed, short-term memory, working memory and executive functions. After 8 weeks, both Group 1 and Group 2 subjects showed significant improvement in LCT, TTA & TTB but improvement in LCT was more marked in Group 1 subjects. Also, there was significant improvement in RDS scores in only Group 1 subjects (P < 0.05). The results thereby, demonstrate that Sahaj Yoga practice in addition to the improvement in various other cognitive domains seen with conventional anti-depressants, can lead to additional improvement in executive functions like manipulation of information in the verbal working memory and added improvement in attention span and visuo-motor speed of the depressives. PMID- 17402268 TI - Wound healing activity of alcoholic extract of Kaempferia galanga in Wistar rats. AB - The wound healing effect of alcoholic extract of Kaempferia galanga (K. galanga) and its effect in dexamethasone suppressed wound healing was studied in Wistar rats. Three wound models viz. incision, excision and dead space wounds were used in this study. The parameters studied were breaking strength in case of incision wounds, epithelialization and wound contraction in case of excision wound and granulation tissue dry weight, breaking strength and hydroxyproline content in case of dead space wound. The dexamethasone treated group showed a significant (P < 0.001) reduction in the wound breaking strength when compared to control group in incision type of wound model. Coadministration of K. galanga with dexamethasone had significantly (P < 0.001) increased the breaking strength of dexamethasone treated group. In excision wound model, the percentage of the wound contraction was significantly (P < 0.05) increased by K. galanga only on 16th day and also it reversed the dexamethasone suppressed wound contraction on the 16 day. K. galanga significantly (P < 0.001) reduced the time required for epithelialization and reversed the epithelialization delaying effect of dexamethasone significantly (P < 0.001). PMID- 17402269 TI - Antioxidant activity of hydroalcoholic leaf extract of ocimum sanctum in animal models of peptic ulcer. AB - In the present study, a hydroalcoholic extract of ocimum sanctum leaves has been investigated for its antioxidant activity in animal models of peptic ulcer with the aim of exploring a possible correlation between its antioxidant and antiulcer activities. Gastric ulcers were produced in rats by ethanol treatment and pyloric ligation whereas duodenal ulcers were produced in guinea pigs by histamine treatment. The animals were divided into six groups of six animals each in all these three models of peptic ulcer. Group I served as diseased control in which distilled water (10 ml/kg) orally was administered as placebo. Group II, III and IV received the test drug (ocimum sanctum leaf extract) in doses of 50 mg/kg, 100 mg/kg and 200 mg/ kg respectively orally once daily for 7 days. Group V was administered ranitidine (10 mg/kg orally) once daily for 7 days and served as standard for comparison. Group VI consisted of healthy control for baseline malondialdehyde (MDA) and superoxide dismutase (SOD) levels. The antioxidant activity was by evaluated estimating plasma MDA in ethanol treated rats and histamine treated guinea pigs and estimating SOD in pyloric ligated rats and histamine treated guinea pigs. In ethanol treated rats, ocimum sanctum leaf extract (100 mg/kg & 200 mg/kg) significantly decreased the levels of MDA to 2.45 +/- 0.29 nmole/ml and 2.40 +/- 0.14 nmole/ml respectively in comparison to 4.87 +/- 0.06 in the diseased control. Similarly, in the histamine treated guinea pig group, the same doses of the extract significantly lowered the levels of MDA to 2.45 +/- 0.12 nmole/ml and 2.37 +/- 0.16 nmole/ml respectively when compared to 4.66 +/- 0.11 in the diseased control. The extract (100 mg/kg & 200 mg/ kg) also increased the levels of SOD in pyloric ligated rats to 1.78 +/- 0.12 U/ml and 1.89 +/- 0.08 U/ml respectively when compared to 1.29 +/- 0.06 U/ml in the diseased control. In the histamine treated guinea pig group also, the same doses of the extract produced a rise in the SOD levels to 2.10 +/- 0.11 U/ml and 2.20 +/- 0.14 U/ml respectively when compared to 1.32 +/- 0.07 in the diseased control. Since lowered levels of MDA and increased levels of SOD signify antioxidant activity, the antiulcer activity of ocimum sanctum might be due to this mechanism. PMID- 17402270 TI - Autonomic status in systemic sclerosis in India. AB - Systemic sclerosis (SSc) is a systemic connective tissue disease characterized by vasomotor disturbances. Autonomic dysfunction has been implicated in the pathogenesis of various symptoms in patients with SSc. In this retrospective study, thirty patients with SSc as cases and thirty age matched healthy subjects were enrolled as controls. The patients as well as controls were subjected to a standard battery of autonomic tests including reactivity and activity (tone) tests. Resting autonomic variables like blood pressure (BP), heart rate (HR) and respiratory rate (RR) were also recorded. In our study, we found significantly higher HR and RR in the patient group (P = 0.0001 and 0.012 respectively). We also observed significantly lower parasympathetic tone and parasympathetic reactivity in SSc patients than the controls. This study shows the status of autonomic functions in SSc patients in India, with lower central parasympathetic drive to the heart. The present study may give some insight for better understanding of this multisystem disorder of unknown etiology. PMID- 17402271 TI - Nonenzymatic glycation of plasma proteins in smokers. AB - Glycation is known to play a key role in complications of many pathophysiological processes. The present study was carried out to assess whether there are abnormalities of nonenzymatic glycation of proteins in smokers. Fourteen current smokers and 10 healthy nonsmokers were enrolled for the present study. Fasting plasma glucose, insulin, fructosamine and total plasma glycated proteins were evaluated. A significant rise in the mean levels of fructosamine and total plasma glycated proteins were found in smokers when compared with controls. Significant difference in insulin values was observed between these two groups. When Pearson's correlation analysis was performed, no significant correlation was found between fasting plasma glucose with either fructosamine or total plasma glycated protein levels. These data suggest an increased glycation of proteins in smokers independent of glucose concentration. PMID- 17402272 TI - Effect of Withania somnifera on forced swimming test induced immobility in mice and its interaction with various drugs. AB - The objective of the present study was to evaluate the antidepressant action of Withania somnifera (WS) as well as its interaction with the conventional antidepressant drugs and to delineate the possible mechanism of its antidepressant action using forced swimming model in mice. Effect of different doses of WS, fluoxetine and imipramine were studied on forced swimming test induced mean immobility time (MIT). Moreover effect of WS 100 mg/kg, i.p. was observed at different time intervals. Effect produced by combination of sub therapeutic doses of WS with imipramine (2.5 mg/kg, i.p.) as well as fluoxetine (2.5 mg/kg, i.p.) were also observed. Effect of WS (100 mg/kg, i.p.) as well as combination of WS (37.5 mg/kg, i.p.) with either imipramine (2.5 mg/kg, i.p.) or fluoxetine (2.5 mg/kg, i.p.) were observed in mice pretreated with reserpine (2 mg/kg, i.p.) and clonidine (0.15 mg/kg, i.p.). Effects of prazosin (3 mg/kg, i.p.) or haloperidol (0.1 mg/kg, i.p.) pre-treatment were also observed on WS induced decrease in MIT. WS produced dose dependent decrease in MIT. Maximum effect in MIT was observed after 30 min of treatment with WS 100 mg/kg, i.p. Combination of WS (37.5 mg/kg, i.p.) with imipramine (2.5 mg/kg, i.p.) or fluoxetine (2.5 mg/kg, i.p.) also produced significant decrease in the MIT. Clonidine and reserpine induced increase in MIT, was significantly reversed by treatment with WS (100 mg/kg, i.p.) as well as combination of WS (37.5 mg/kg, i.p.) with either imipramine (2.5 mg/kg, i.p.) or fluoxetine (2.5 mg/kg, i.p.). Pre-treatment with prazosin but not haloperidol, significantly antagonized the WS (100 mg/kg, i.p.) induced decrease in MIT. It is concluded that, WS produced significant decrease in MIT in mice which could be mediated partly through a adrenoceptor as well as alteration in the level of central biogenic amines. PMID- 17402273 TI - Antioxidant response to oxidative stress induced by smoking. AB - Oxygen free radicals have been hypothesized to play a pivotal role in the deleterious effects of smoking on health. The present study was undertaken to examine the oxidant and antioxidant system among smokers and nonsmokers. Fourteen smokers and 11 nonsmokers were enrolled for this study. The protein carbonyl levels in smokers were found to be significantly higher than in nonsmokers. The levels of plasma ascorbic acid, free sulfhydryl group, and erythrocyte reduced glutathione were lower in smokers compared to nonsmokers. In smokers the erythrocyte activities of both glutathione peroxidase and catalase were decreased when compared to that in nonsmokers. The data from the study reemphasizes the presence of oxidative stress in smokers. The concomitant decrease in the activities of both catalase and glutathione peroxidase found in the erythrocytes of smokers raises rational grounds for expressing concern over the increased susceptibility towards oxidative stress in these subjects. PMID- 17402274 TI - Efficacy and tolerability of trandolapril in mild to moderate hypertension--a double blind comparative clinical trial with enalapril in Indian population. AB - Several large scale clinical trials have demonstrated that angiotensin converting enzyme inhibitors offer cardiovascular and renal protection independent of their effects on systolic BP. Trandolapril is a new angiotensin converting enzyme inhibitor approved for the treatment of hypertension. The potential advantages of this drug are long duration of action and better tolerability. The objective of the study was to compare the efficacy and tolerability of trandolapril with that of enalapril in mild to moderate hypertension in Indian population. In this double blind, multicentric, parallel comparative clinical study, 120 patients with mild to moderate hypertension were randomly assigned to receive trandolapril 2 mg or enalapril 5 mg once daily for 8 weeks. The attainment of sitting diastolic blood pressure <90 mmHg at the end of 8th week was considered as primary outcome measure and attainment of diastolic blood pressure <90 mmHg or reduction of at least 10 mmHg diastolic blood pressure compared to baseline at any visit was considered as secondary outcome measures. 98.4% patients treated with trandolapril and 92.6% patients treated with enalapril fulfilled the primary outcome measure. 54, 72 and 62% patients on trandolapril and 52, 61 & 64% patients on enalapril fulfilled secondary outcome measure at the end of 2nd, 4th and 8th week respectively. Also trandolapril was better tolerated than enalapril with no significant abnormality in lab parameters. PMID- 17402275 TI - A reverse phase high performance liquid chromatography method for simultaneous estimation of melatonin, carbamazepine epoxide and carbamazepine simultaneously in serum. AB - Monitoring of plasma antiepileptic drugs is useful for better clinical management in epileptic patients, particularly in children. Carbamazepine (CBZ) is one of the commonly prescribed anticonvulsants. The active metabolite of carbamazepine carbamazepine-10-11 epoxide (CBZ-Epo) also exhibits anticonvulsant effect. The pineal hormone, melatonin exerts an anticonvulsant effect in experimental seizure models and recently has also been used in cases of childhood epilepsy. To facilitate the simultaneous plasma estimation of carbamazepine, carbamazepine epoxide, and melatonin, a new HPLC method was developed. Waters millennium 2010 chromatography manager with a 515 HPLC pump and Waters 24879 dual absorbance UV detector was used. A 25 microlitre of sample and standards were injected, and chromatographic separation was achieved by Merck C18 reverse phase column particle size 5 micro, 250 mm x 4 mm. It was quantitated at UV light 210 nm. The retention times of melatonin, CBZ-Epo, and CBZ were 6.3 min, 7.5 min, and 13.9 min respectively. The Mobile Phase used was water: acetonitrile (70:30), pH 3.0 adjusted with orthophosphoric acid at the flow rate of 1 ml/min. The limits of detection of melatonin, carbamazepine epoxide, and carbamazepine were 800, 500, and 1300 pg respectively. PMID- 17402276 TI - Antihyperglycemic effects of newly synthesized 6,7-bis[2-(substitutedphenyl)-4 oxothiazolidin3yl1 quinoxaline 2,3-(1 FH, 4 H)-diones. PMID- 17402277 TI - Waterborne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt. AB - At least 325 water-associated outbreaks of parasitic protozoan disease have been reported. North American and European outbreaks accounted for 93% of all reports and nearly two-thirds of outbreaks occurred in North America. Over 30% of all outbreaks were documented from Europe, with the UK accounting for 24% of outbreaks, worldwide. Giardia duodenalis and Cryptosporidium parvum account for the majority of outbreaks (132; 40.6% and 165; 50.8%, respectively), Entamoeba histolytica and Cyclospora cayetanensis have been the aetiological agents in nine (2.8%) and six (1.8%) outbreaks, respectively, while Toxoplasma gondii and Isospora belli have been responsible for three outbreaks each (0.9%) and Blastocystis hominis for two outbreaks (0.6%). Balantidium coli, the microsporidia, Acanthamoeba and Naegleria fowleri were responsible for one outbreak, each (0.3%). Their presence in aquatic ecosystems makes it imperative to develop prevention strategies for water and food safety. Human incidence and prevalence-based studies provide baseline data against which risk factors associated with waterborne and foodborne transmission can be identified. Standardized methods are required to maximize public health surveillance, while reporting lessons learned from outbreaks will provide better insight into the public health impact of waterborne pathogenic protozoa. PMID- 17402278 TI - Health risks in wastewater irrigation: comparing estimates from quantitative microbial risk analyses and epidemiological studies. AB - The combination of standard quantitative microbial risk analysis (QMRA) techniques and 10,000-trial Monte Carlo risk simulations was used to estimate the human health risks associated with the use of wastewater for unrestricted and restricted crop irrigation. A risk of rotavirus infection of 10(-2) per person per year (pppy) was used as the reference level of acceptable risk. Using the model scenario of involuntary soil ingestion for restricted irrigation, the risk of rotavirus infection is approximately 10(-2) pppy when the wastewater contains < or =10(6) Escherichia coli per 100ml and when local agricultural practices are highly mechanised. For labour-intensive agriculture the risk of rotavirus infection is approximately 10(-2) pppy when the wastewater contains < or = 10(5) E. coli per 100ml; however, the wastewater quality should be < or = 10(4) E. coli per 100ml when children under 15 are exposed. With the model scenario of lettuce consumption for unrestricted irrigation, the use of wastewaters containing < or =10(4) E. coli per 100ml results in a rotavirus infection risk of approximately 10(-2) pppy; however, again based on epidemiological evidence from Mexico, the current WHO guideline level of < or =1,000 E. coli per 100ml should be retained for root crops eaten raw. PMID- 17402279 TI - Health impact of water and sanitation infrastructure reconstruction programmes in eight Central American communities affected by Hurricane Mitch. AB - In response to Hurricane Mitch, which struck Central America in October-November 1998, the American Red Cross (ARC) and the Centers for Disease Control and Prevention (CDC) collaborated on a 3-year evaluation of the public health impact of ARC's water, sanitation and hygiene education activities in eight study areas in Honduras, Nicaragua, El Salvador and Guatemala. The evaluation compared: 1) access to and use of water and sanitation facilities, 2) the use of hygienic behaviours, and 3) diarrhoeal prevalence in children younger than 3 years of age before (February 2000) and after (February 2002) the interventions had been implemented. The evaluation included household and key informant interviews designed to measure these three components. Water quality of community water sources and household water was evaluated by measuring levels of indicator bacteria. During the final survey, an infrastructure evaluation provided a review of the design, construction, and current operation and maintenance of the water systems and latrines. The integrated water and sanitation infrastructure interventions and hygiene education programmes implemented following Hurricane Mitch effectively decreased diarrhoea prevalence in the target communities. PMID- 17402280 TI - Disease burden estimation to support policy decision-making and research prioritization for arsenic mitigation. AB - The main response to arsenic contamination of shallow tubewells in Bangladesh is the provision of alternative water supplies. To support decision-making in relation to alternative water supply selection, the Arsenic Policy Support Unit commissioned the development of a tool for estimating disease burdens for specific options using disability-adjusted life years as the metric. This paper describes the assumptions in dose-responses, relationships between microbial indicators and pathogens, water consumed and population characteristics used, and presents a case study of how the tool was applied. Water quality data and dose response models were used to predict disease burdens due to microbial pathogens and arsenic. Disease burden estimates predicted by the tool were based on evidence in the published literature. There were uncertainties in key assumptions of water consumed and the ratio of microbial indicators and pathogens, which led to broad confidence intervals and the need to consider the results in a wider context and further research needs. Deep tubewells and rainwater harvesting had the lowest disease burden estimates, while pond sand filters and dug wells had much higher predicted disease burden due to frequent microbial contamination. The need for rigorous water supply protection through water safety plans was highlighted. At present, the risk assessment is useful for informing judgement by experienced water and health professionals and identifying key research questions. Improved arsenic dose-response models and a better understanding of the relationship between microbial indicators and pathogens in tropical settings are required. PMID- 17402281 TI - Field scale quantification of microbial transport from bovine faeces under simulated rainfall events. AB - The dispersion and transport of Cryptosporidium parvum oocysts, Escherichia coli and PRD1 bacteriophage seeded into artificial bovine faecal pats was studied during simulated rainfall events. Experimental soil plots were divided in two, one sub-plot with bare soil and the other with natural vegetation. Simulated rainfall events of 55 mm.h(-1) for 30 min were then applied to the soil plots. Each experimental treatment was performed in duplicate and consisted of three sequential artificial rainfall events ('Runs'): a control run (no faecal pats); a fresh faecal pat run (fresh faecal pats); and an aged faecal pat run (one week aged faecal pats). Transportation efficiency increased with decreasing size of the microorganism studied; Cryptosporidium oocysts were the least mobile followed by E. coli and then PRD1 phage. Rainfall events mobilised 0.5 to 0.9% of the Cryptosporidium oocysts, 1.3-1.4% of E. coli bacteria, and 0.03-0.6% of PRD1 bacteriophages from the fresh faecal pats and transported them a distance of 10 m across the bare soil sub-plots. Subsequent rainfall events applied to aged faecal pats only mobilised 0.01-0.06% of the original Cryptosporidium oocyst load, between 0.04 and 15% of the E. coli load and 0.0006-0.06% of PRD1 bacteriophages, respectively. PMID- 17402282 TI - Larval habitats of potential mosquito vectors of West Nile virus in the Florida Keys. AB - The occurrence of larvae of two potential vectors of West Nile virus, Culex nigripalpus and Culex quinquefasciatus, was examined in the Florida Keys. About half of the aquatic habitats examined contained larvae of either one or both of the species. Culex quinquefasciatus was the most frequently encountered species, whereas only 9% of habitats sampled contained Culex nigripalpus. Over half of those samples that contained Culex nigripalpus also contained Culex quinquefasciatus. The two species utilize similar larval habitats in the Florida Keys, Monroe County, Florida, USA. PMID- 17402283 TI - Assessment of the chemical quality of drinking water in Cambodia. AB - Most water supply programmes in Cambodia have focused on providing access to bacteriologically safe water, an approach which has led to an increasing reliance on ground water, especially in rural areas. However, there have been very few data collected on the chemical quality of the nation's drinking water sources, and few water supply programmes have the capacity to assess chemical quality. The study was designed to address this data gap by conducting a low-cost, rapid assessment of drinking water sources nationwide to determine whether there were any chemicals of concern in Cambodian water supply sources. Results of the assessment confirm that there are several parameters of health and aesthetic concern; dissolved arsenic is the most significant. Elevated arsenic levels (some exceeding 500 microgl(-1)) were detected in aquifers of moderate depth in several highly populated areas, confirming that further investigation of the occurrence of arsenic contamination in Cambodia is warranted. Other chemicals of health concern include nitrate, nitrite, fluoride and manganese. Additionally, many ground water sources are negatively impacted by parameters of aesthetic concern, such as iron, manganese, hardness and total dissolved solids. Elevated levels of these parameters have caused consumers to reject newly installed water supplies, often in favour of surface water sources that are bacteriologically unsafe. PMID- 17402284 TI - Microbial risk assessment of local handling and use of human faeces. AB - Dry urine-diverting toilets may be used in order to collect excreta for the utilisation of nutrients. A quantitative microbial risk assessment was conducted in order to evaluate the risks of transmission of infectious disease related to the local use of faeces as a fertiliser. The human exposures evaluated included accidental ingestion of small amounts of faeces, or a mixture of faeces and soil, while emptying the storage container and applying the material in the garden, during recreational stays to the garden, and during gardening. A range of pathogens representing various groups of microorganisms was considered. Results showed that 12-months' storage before use was sufficient for the inactivation of most pathogens to acceptable levels. When working or spending time in the garden the annual risk of infection by Ascaris was still slightly above 10(-4) in these scenarios, although the incidence rate for Ascaris is very low in the population in question. Measures to further reduce the hygienic risks include longer storage, or treatment, of the faeces. The results can easily be extended to other regions with different incidence rates. PMID- 17402285 TI - Unsafe tap water in households supplied from groundwater in the Salento Region of Southern Italy. AB - Although the fractured aquifer of the Salento supplies over 80% of the drinking water requirements of the local population, its exposure to pollution has recently increased. In recent years, owing to the arid climate and droughts, the spreading of wastewater on soil for irrigation has become much more frequent. Consequently, hazardous and pathogenic microorganisms released with wastewater have been transported into the subsoil and have contaminated groundwater. An elaboration of epidemiological data has shown that the local population has the highest exposure to endemic gastroenteritis in Italy. In order to reduce human exposure to unsafe groundwater, the setback distance for drinking wells necessary to achieve the 'natural disinfection' criteria, has been determined experimentally at the Nardo aquifer (Salento region), supported by groundwater monitoring results and a mathematical transport model able to determine the apparent pathogenic microorganism pathways in fractures. The results also provided valuable inactivation constants of cultural indicators (coliforms, enterococci, Clostridium spores and somatic coliphages) and viruses in the wastewater that have been injected into the fractured aquifer since 1991. Furthermore, the efficacy of chlorine to remove viral indicators from water in a well 500 m from wastewater injection was tested. Hypochlorination reduces somatic coliphages and Clostridium spores in groundwaters but did not achieve complete inactivation in all tests. Complete disinfection of groundwater samples was possible only when there was an initial Clostridium spores count of < or = 10 CFU 100 ml (-1). PMID- 17402286 TI - Waterborne pathogens in urban watersheds. AB - A serious concern for managers of water resources, pathogens in the urban environment easily enter waters through a number of pathways, including discharge of inadequately treated sewage, stormwater runoff, combined sewer overflows and sanitary sewer overflows. Pathogens in US ambient water bodies are regulated under the Clean Water Act (CWA), while pathogens in drinking water supplies are regulated under the Safe Drinking Water Act. Total maximum daily loads (TMDLs) are developed in accordance with CWA regulations for ambient water bodies with bacterial concentrations exceeding the water quality standard, which generally is a measure of a bacterial indicator organism. However, developing a TMDL for a supplementary indicator or pathogen is also required if a use impairment would still exist even after the water body is in compliance with the standard. This occurs because indicator organisms do not reflect the presence of pathogen contamination with complete certainty. The evaluation of pathogen indicators and summary of epidemiological studies presented are resources for those developing TMDLs to achieve water quality standards and restore water bodies to their intended uses. PMID- 17402287 TI - Occurrence of Cryptosporidium oocysts and Giardia cysts in the Nakdong River and their removal during water treatment. AB - This study was conducted in preparation of a pending Cryptosporidium regulation in Korea. The study had two main objectives: 1) to examine the occurrence of Cryptosporidium oocysts in the Nakdong River; and 2) to evaluate their removal during water treatment. Occurrence of Giardia cysts was also examined. Average (arithmetic mean) numbers of Cryptosporidium oocysts and Giardia cysts at the treatment intake site were 2.61(-1) and 4.81(-1), respectively. Generally, the number of Giardia cysts was higher than that of Cryptosporidium oocysts at more sites, but the difference was minimal. Comparison of tributaries indicated that livestock wastes were more serious pollutants than sewage in terms of protozoa contamination. In general, fewer oocysts and cysts were detected during winter. No correlation was found for such water quality parameters as T-N, T-P, TOC, DO, pH and temperature with the numbers of oocysts and cysts except for suspended solids, which showed the highest correlation (R2 = 0.55). Removal of Cryptosporidium oocysts was evaluated using a Cryptosporidium tracer, which has similar characteristics to Cryptosporidium oocysts. The tracer removal depended on turbidity removal. Coagulation followed by sedimentation resulted in 1.2-1.5 log removal of the tracer under optimal conditions. Filtration resulted in 1.3 1.5 log removal of the tracer. These treatability experiments showed that traditional water treatment processes could achieve 2.5-3.0 log removal of the oocysts. PMID- 17402288 TI - Microbial counts and pesticide concentrations in drinking water after alum flocculation of channel feed water at the household level, in Vinh Long Province, Vietnam. AB - Low technology water supply, treatment and sanitation techniques were reviewed in a Vietnamese village in the Mekong River Delta as part of an AusAID Project to reduce poverty and to improve the overall living standards and health in the Cuu Long (Mekong) delta for 500,000 rural poor. It was found that dosing of canal water in 200-1 ceramic jars with alum, PAC or PAC followed by hypochlorite provides a high level of treatment for drinking water at the household level in a rural village setting in the Mekong River delta. Seventy per cent (7 in total) of the 24 samples collected showed that Escherichia coli levels per 100 ml in the jars after treatment showed a 'no risk' profile (0 E. coli 100 ml(-1)) i.e. the water would be considered to be of very good quality for drinking according to World Health Organization Guidelines for Rural Drinking Water Supply. This included the eight samples that were dosed with hypochlorite. The remaining 30% of samples (seven in total) ranged from 14 to 47 E. coli 100 ml(-1) which is considered an intermediate risk (10-100 E. coli 100 ml(-1)) according to WHO guidelines. All control water samples (eight in total) were high or very high risk (>100 E. coli 100ml(-1). PMID- 17402289 TI - The health-related microbiological quality of bottled drinking water sold in Dar es Salaam, Tanzania. AB - The consumption of bottled and plastic-bagged drinking water in Tanzania has increased largely because of the deteriorating quality of tap water. It is uncertain whether these water products are safe for drinking. In this study, the microbiological quality of bottled and plastic-bagged drinking water sold in Dar es Salaam, Tanzania, was investigated. One hundred and thirty samples representing 13 brands of bottled water collected from shops, supermarkets and street vendors were analysed for total coliform and faecal coliform organisms as well as heterotrophic bacteria. These were compared with 61 samples of tap water. Heterotrophic bacteria were detected in 92% of the bottled water samples analysed. Total and faecal coliform bacteria were present in 4.6% and 3.6%, respectively, of samples analysed with a tendency for higher contamination rates in plastic-bagged drinking water. Microbiological quality of tap water was found to be worse compared with bottled water, with 49.2% and 26.2% of sampling points showing the presence of total coliform and faecal coliform organisms, respectively. The results suggest caution and vigilance to avert outbreaks of waterborne diseases from these types of drinking water. PMID- 17402290 TI - Concentration, compartmentation and metabolic function of intracellular free Mg2+. AB - Intracellular total Mg2+ and free Mg2+ are compartmentalized between cell organelles and within the cytosol. Different values of [Mg2+]i in the cytosol of the same cell type were measured by various investigators. A main reason for the differences is the uncertainty of the dissociation constants used for the Mg furaptra complex in the fluorescence method and for MgATP when 31P NMR was employed. The more realistic KD values of Mg furaptra and MgATP measured under in situ conditions are higher than the KDs used by most investigators. The [Mg2+]is obtained and the KDs used by various authors were presented. The role of intracellular Mg2, in metabolic functions and the action of various effectors on [Mg2+]i and [Ca2+] was reviewed. Intracellular Mg2+ may have a permissive role supporting the effector-induced mechanisms that are mediated by Ca2+ as a second messenger. PMID- 17402291 TI - High fructose consumption combined with low dietary magnesium intake may increase the incidence of the metabolic syndrome by inducing inflammation. AB - The metabolic syndrome is a cluster of common pathologies: abdominal obesity linked to an excess of visceral fat, insulin resistance, dyslipidemia and hypertension. This syndrome is occurring at epidemic rates, with dramatic consequences for human health worldwide, and appears to have emerged largely from changes in our diet and reduced physical activity. An important but not well appreciated dietary change has been the substantial increase in fructose intake, which appears to be an important causative factor in the metabolic syndrome. There is also experimental and clinical evidence that the amount of magnesium in the western diet is insufficient to meet individual needs and that magnesium deficiency may contribute to insulin resistance. In recent years, several studies have been published that implicate subclinical chronic inflammation as an important pathogenic factor in the development of metabolic syndrome. Pro inflammatory molecules produced by adipose tissue have been implicated in the development of insulin resistance. The present review will discuss experimental evidence showing that the metabolic syndrome, high fructose intake and low magnesium diet may all be linked to the inflammatory response. In many ways, fructose-fed rats display the changes observed in the metabolic syndrome and recent studies indicate that high-fructose feeding is associated with NADPH oxidase and renin-angiotensin activation. The production of reactive oxygen species results in the initiation and development of insulin resistance, hyperlipemia and high blood pressure in this model. In this rat model, a few days of experimental magnesium deficiency produces a clinical inflammatory syndrome characterized by leukocyte and macrophage activation, release of inflammatory cytokines, appearance of the acute phase proteins and excessive production of free radicals. Because magnesium acts as a natural calcium antagonist, the molecular basis for the inflammatory response is probably the result of a modulation of the intracellular calcium concentration. Potential mechanisms include the priming of phagocytic cells, the opening of calcium channels, activation of N-methyl-D-aspartate (NMDA) receptors, the activation of nuclear factor-kappaB (NFkB) and activation of the renin-angiotensin system. Since magnesium deficiency has a pro-inflammatory effect, the expected consequence would be an increased risk of developing insulin resistance when magnesium deficiency is combined with a high-fructose diet. Accordingly, magnesium deficiency combined with a high-fructose diet induces insulin resistance, hypertension, dyslipidemia, endothelial activation and prothrombic changes in combination with the upregulation of markers of inflammation and oxidative stress. PMID- 17402292 TI - Magnesium therapy in acoustic trauma. AB - Acoustic trauma is one of the major causes of hearing loss and tinnitus, particularly in industrial environments. Noise-induced hearing loss (NIHL) results in direct mechanical damage as well as in indirect metabolic processes. Metabolic disorders have multiple origins: ionic, ischemic, excitotoxic and production of cochlear free radicals causing cell death, due to necrosis or apoptosis. The efficacy of magnesium, administered either to prevent or to treat NIHL has been demonstrated in several studies in animals and in humans. Magnesium, which easily crosses the hematocochlear barrier, presents neuroprotective and vasodilatory effects, and thus, is able to limit the cochlear damage. Magnesium therapy is well documented because it is usually prescribed in other pathologies. Its side effects and contraindications are few and it is cheap. This article presents also some arguments that emphasize the interest of magnesium therapy in acoustic trauma. PMID- 17402293 TI - Onset of nephrocalcinosis depends on dietary phosphorus concentration in male rats fed a magnesium-deficient diet. AB - Although a magnesium (Mg)-deficient diet is generally known to induce nephrocalcinosis, our previous study observed that despite the administration of a Mg-deficient diet, the kidney calcium (Ca) and phosphorus (P) concentrations were not increased in male rats. We speculated that this result was due to the P concentration of the experimental diet based on the AIN-93G formula used in the previous study. In the present study, male rats were fed modified AIN-93G diets containing the two different Mg concentrations [0.5 g per kg diet (normal-Mg) or Mg-free (Mg-deficient)] and three different P concentrations [3 (3-P), 5 (5-P) or 7 (7-P) g per kg diet]. By histological examination of the kidney, nephrocalcinosis was not observed in rats fed on the Mg-deficient diet containing 3-P While nephrocalcinosis appeared in rats fed on the Mg-deficient diet containing 5-P and 7-P The degree of nephrocalcinosis was severe in rats fed on the Mg-deficient diet containing 7-P compared with rats fed on the Mg-deficient diet containing 5-P These results demonstrated that the Mg-deficient diet based on AIN-93G formula dose not induce nephrocalcinosis and that the Mg-deficient diet based on AIN-93G formula with increased dietary P concentrations induces nephrocalcinosis in male rats. We suggest that the onset of nephrocalcinosis could depend on the dietary P concentration in male rats fed on a Mg-deficient diet. PMID- 17402294 TI - Blood magnesium concentration and dopamine or dobutamine infusion demand in patients during CABG (coronary artery bypass grafting) with normovolemic haemodilution. AB - It is well known that magnesium (Mg) plays an essential role in cardiac protection. Mg has many beneficial effects on the myocardium and cardiac function, e.g. it improves contractility and reduces the number of cardiac arrhythmia episodes. The inotropically positive effects of Mg are interesting and worth stressing. High blood Mg concentration may result in an increase in cardiac contraction strength, which may be important for haemodynamic stabilization, and thus it is likely to decrease the demand for dopamine and dobutamine infusions. However, the exact determination of correlation between blood Mg concentrations and dopamine or dobutamine infusion demand is still unknown. The aim of the study was to assess the demand for dopamine or dobutamine infusion in relation to changes in blood magnesium concentrations in patients undergoing CABG (Coronary artery bypass graft) with extracorporeal circulation and normovolemic haemodilution. The study included 20 male patients, aged 53-70 (61.1 +/- 6.9) who underwent general anaesthesia and coronary artery bypass grafting (CABG) with extracorporeal circulation (ECC) and normovolemic haemodilution (NH) due to stable angina pectoris. The patients were retrospectively divided into three groups: A--patients who did not receive dopamine or dobutamine infusion, B--those receiving only D infusion in the doses dependent on their clinical state and C- those receiving DB infusion in the doses dependent on their clinical state. Mg was measured in 7 stages: 1) just before anaesthesia after the radial artery cannulation, 2) during normovolemic haemodilution and ECC, 3) immediately after surgery, 4) in the evening of the surgery day, 5) in the morning of the lst postoperative day, 6) in the evening of 1st postoperative day, 7) in the morning of the 2nd postoperative day. The spectrophotometric methods were used to determine Mg. The CABG procedure resulted in a decrease in Mg. Its level returned to normal in the evening of surgery day. The NH caused a similar Mg decrease in groups A, B and C, but these significantly low values of Mg were observed only in stage 2. There was no correlation between blood Mg concentrations and dopamine or dobutamine infusion demand. IN CONCLUSION: 1) The CABG procedure resulted in decreased blood magnesium concentrations. 2) The Mg changes do not correlate with dopamine or dobutamine infusion demand. PMID- 17402295 TI - Magnesium intake is inversely associated with the prevalence of tooth loss in Japanese pregnant women: the Osaka Maternal and Child Health Study. AB - There have only been a few studies on the role of mineral intake in tooth loss. We investigated the association between mineral intake and the prevalence of tooth loss in Japan. We used the baseline data on 1002 pregnant women who were enrolled in the Osaka Maternal and Child Health Study between November 2001 and March 2003. Tooth loss was defined as the previous extraction of one or more teeth. Nutrient intake was assessed by a validated diet history questionnaire. Prevalence odds ratios and confidence intervals were estimated by applying a multiple logistic regression model. The adjusted odds ratio upon comparison of the highest quartile with the lowest quartile of magnesium intake was 0.64 (95% confidence interval, 0.42-0.99), showing a tendency for an inverse dose-response relationship (p for linear trend = 0.05). There were no associations between the level of consumption of calcium, phosphate, iron, zinc, or copper and tooth loss. The present findings suggest that intake of magnesium is related to reduced prevalence of tooth loss among young Japanese women. PMID- 17402296 TI - Dead regions in the cochlea at 4 kHz in elderly adults: relation to absolute threshold, steepness of audiogram, and pure-tone average. AB - The aims of this study were (1) to investigate the prevalence of dead regions (DRs) at 4 kHz in elderly people with hearing loss and (2) to determine the extent to which the presence/absence of a DR can be predicted from the absolute threshold, the slope of the audiogram, or the pure-tone average (PTA) hearing loss at 0.5, 1, and 2 kHz. DRs were assessed for 98 ears with absolute thresholds between 60 and 85 dB HL at 4 kHz using the threshold equalizing noise test. Thirty-six ears had a DR at 4 kHz. There was no statistically significant difference in the slope of the audiogram or PTA between ears with and without DRs. However, the mean absolute threshold at 4 kHz was significantly higher for the group with DRs than for the group without DRs. The prevalence of DRs exceeded 50% for hearing losses greater than 70 dB. PMID- 17402297 TI - Effects of stimulus level on cortical auditory event-related potentials evoked by speech. AB - The effect of stimulus level on cortical auditory event-related potentials (ERPs) evoked by consonant-vowel (CV) contrasts, /ta/, /da/, and /sa/, was investigated. The lowest level at which CVs were discriminated with >95% accuracy was determined for 15 normally hearing adults. ERPs were obtained at 0, 20, and 40 dB SL above this level during active listening. ERP latencies decreased as level increased. P300 amplitude did not vary with CV level or type; however, obligatory ERPs decreased in amplitude as level increased. The effect of level on P300 latency is likely related to the cognitive processing speed needed to perform speech discrimination. Obligatory ERP amplitude results suggest that attention demands vary with level during discrimination of speech features. PMID- 17402298 TI - The relationship between obligatory cortical auditory evoked potentials (CAEPs) and functional measures in young infants. AB - Finding ways to evaluate the success of hearing aid fittings in young infants has increased in importance with the implementation of hearing screening programs. Cortical auditory evoked potentials (CAEP) can be recorded in infants and provides evidence for speech detection at the cortical level. The validity of this technique as a tool of hearing aid evaluation needs, however, to be demonstrated. The present study examined the relationship between the presence/absence of CAEPs to speech stimuli and the outcomes of a parental questionnaire in young infants who were fitted with hearing aids. The presence/absence of responses was determined by an experienced examiner as well as by a statistical measure, Hotelling's T(2). A statistically significant correlation between CAEPs and questionnaire scores was found using the examiner's grading (rs = 0.45) and using the statistical grading (rs = 0.41), and there was reasonably good agreement between traditional response detection methods and the statistical analysis. PMID- 17402299 TI - Can the acoustic change complex be recorded in an individual with a cochlear implant? Separating neural responses from cochlear implant artifact. AB - The purpose of this case study was to determine whether the P1-N1-P2 acoustic change complex (ACC) could be recorded in an individual with a cochlear implant. In a cochlear implant recipient, stimulus-related artifact from the implant can overlap the evoked potential of interest, making it difficult to determine whether the recorded response is neural or a simple reflection of the artifact. This is an even greater technical challenge for the ACC because stimuli having relatively long durations are used. The subject was a 24-year-old with a diagnosis of auditory neuropathy/auditory dys-synchrony and used a MED-EL Tempo+ cochlear implant in her left ear. The ACC was recorded to synthetic vowels containing a change of F2 at midpoint ranging from 0 (no change) to 1200 Hz (perceived as /ui/). The stimuli were presented randomly at 75 dB SPL via a loudspeaker. In one condition the subject ignored the stimuli and watched a captioned video. In the other, the subject pressed one button on a response pad if she perceived an acoustic change at stimulus midpoint and another if she did not. Cortical auditory evoked potentials were recorded from 32 scalp electrodes. Results indicated that the ACC was present and could be teased apart from the cochlear implant stimulus artifact. ACC thresholds showed good agreement with behavioral discrimination performance, and therefore, results are positive for the potential clinical application of the ACC technique to individuals with cochlear implants. PMID- 17402300 TI - The effect of presentation level on the Gaps-In-Noise (GIN) test. AB - The Gaps-In-Noise (GIN) test is a new procedure used in the diagnosis of central auditory processing disorders. Performance on the GIN is recorded as approximate gap detection threshold and percent correct. In order to utilize the GIN test clinically, it is important to know how presentation level influences performance on the GIN. To this end, ten normal-hearing adults were administered the GIN at 5, 10, 15, 20, 25, 30, 35, and 50 dB SL with regard to threshold to GIN noise. Results indicated that performance for both the approximate gap detection threshold (A.th) and percent correct improved with increasing presentation level. Performance at 35 dB SL was not significantly different from the standard clinical presentation level (50 dB SL). Gaps that were between 5 and 8 msec in duration tended to show more variation across presentation levels. Although an influence of presentation level was noted, this influence should not be manifested at the standard clinical presentation level. PMID- 17402301 TI - A systematic review of health-related quality of life and hearing aids: final report of the American Academy of Audiology Task Force On the Health-Related Quality of Life Benefits of Amplification in Adults. AB - This is the final report of the American Academy of Audiology Task Force on the Health-Related Quality of Life (HRQoL) Benefits of Amplification in Adults. A systematic review with meta-analysis examined evidence pertaining to the use of hearing aids for improving HRQoL for adults with sensorineural hearing loss (SNHL). Relevant search strings applied to the CENTRAL, CINAHL, Cochrane Reviews, ComDisDome, EBMR, and PubMed databases identified randomized controlled trial, quasi-experimental, and nonexperimental pre-post test designed studies. Sixteen studies met a priori criteria for inclusion in this review. A random-effects meta analysis showed differential results for generic versus disease-specific HRQoL measures for within- and between-subject designs. Although generic measures used for within-subject designs did not demonstrate HRQoL benefits from hearing aids, mean effect sizes and confidence intervals for within-subject designs and disease specific instruments suggested that hearing aids have a small-to-medium impact on HRQoL. Further, the between-subject studies supported at least a small effect for generic measures, and when measured by disease-specific instruments, hearing aids had medium-to-large effects on adults' HRQoL. This review concludes that hearing aids improve adults' HRQoL by reducing psychological, social, and emotional effects of SNHL. Future studies should include control groups using randomized controlled trials. PMID- 17402302 TI - Further questions about the acceptable noise level test. PMID- 17402303 TI - Usefulness of acoustic reflexes still unproven in newborns and young infants. PMID- 17402304 TI - Where does the responsibility for health governance in the UK lie? PMID- 17402305 TI - Policy and politics. PMID- 17402306 TI - Can we fix it. Interview by Mandy Murphy. PMID- 17402307 TI - Impact of a traffic light nutrition tool in a primary school. PMID- 17402308 TI - Don't kill me, don't keep me waiting, and don't waste resources. PMID- 17402309 TI - Public-private partnerships: an old solution to present challenges. PMID- 17402310 TI - Health, wealth and the pursuit of happiness. PMID- 17402311 TI - Planning for health: bringing the professions together. PMID- 17402312 TI - Health, spending and the effort to improve quality in OECD countries: a review of the data. AB - The performance of health systems is of concern to both policy-makers and academics and a large body of recent literature has advanced the debate significantly on methods and results of health system performance assessments. In this article, I attempt to summarize what is known about a range of areas of health system performance, specifically in the areas of spending and outcomes, using data from the Organization for Economic Cooperation and Development (OECD) in the areas of health, spending, risk factors and quality of care. In so doing, we use new data from the OECD's Health Care Quality Indicators (HCQI) to examine a factor that is frequently cited at the national level but rarely compared at the international level, namely the quality of the healthcare provided. In keeping with other assessments of trends in health, we show that health has improved dramatically since the 1970s in all of the countries of the OECD. Likewise, all of the OECD countries are spending many times more on health per person than in 1970. However, the gains in health as well as the spending levels vary tremendously across countries. Quality of care is relatively high in some areas such as vaccination rates. In other areas such as breast cancer survival, the HCQI data show that most countries are making progress somewhat universally, but that all countries still have progress to be made. Finally, in other areas, such as inpatient care for acute myocardial infarction (AMI), there is wide variation in quality. Future work will need to be undertaken in order to examine possible best practices at the policy and operational level of health care delivery, their impact on spending and, most importantly, on health outcomes. PMID- 17402313 TI - The future of regulation and governance. AB - The NHS in England is undergoing considerable structural change as it introduces more locally accountable hospitals, greater patient choice, payment by results and local general practitioner based commissioning of services. To deliver the anticipated service improvement associated with these changes, there is a need to ensure that the reformed NHS provides incentives for service improvement, while at the same time maintaining consistency in access to services and the safe delivery of services. The UK government is committed to reducing bureaucracy, which is based upon the associated desire to reduce the overall costs of regulation, while increasing the accountability of health services. This requires a new approach to regulation, balancing a standardised concept of quality with incentives to encourage local innovation in service design and delivery. Accountability through regulation brings with it the burdens of supplying information for audit and inspection, which are placed upon the organization. This article examines the issues that need to be considered in the design of an effective regulatory system; one that must operate within a restricted cost envelope, and in particular focuses on the concept of reducing the administrative burden associated with both inspection and self-assessment and the need to make these proportionate to a demonstrable contribution to the improvement in regulation and the ultimate goal of improvement in quality of healthcare. PMID- 17402314 TI - Where is the pilot? The changing shapes of governance in the European hospital sector. AB - Hospital governance refers to the complex of checks and balances that determine how decisions are made within the top structures of hospitals. This article explores the essentials of the concept by analysing the root notion of governance and comparing it with applications in other sectors. Recent developments that put pressure on the decision-making system within hospitals are outlined. Examples from the UK, France and the Netherlands are presented. Based on an evaluation of the current state of affairs, a research framework is developed, focusing on the determinants of governance configurations within the national healthcare systems and the wider legal and socio-economic context, as well as on the impact of governance configurations on the efficiency of the governing bodies and overall hospital performance. The article concludes with a preview of the European Hospital Governance Project, which follows the outlines of the described research framework. New techniques of data mining that are used in this project are explained by means of a real data example. PMID- 17402315 TI - The problem of engaging hospital doctors in promoting safety and quality in clinical care. AB - There is widespread agreement that the medical profession has much to learn about addressing adverse events in clinical practice and participating in clinical governance. In England and Wales centrally driven initiatives such as medical audit, clinical governance and the National Reporting and Learning System have failed to transform the management of iatrogenic adverse events. In this article we explore the historical and cultural background of these issues with respect to hospital medicine and suggest means of tackling the challenges ahead. PMID- 17402316 TI - Out-of-control tort system has far-reaching effects. PMID- 17402317 TI - Association of environmental lead exposure with academic performance: Louisville, Kentucky, 2003. PMID- 17402318 TI - Practice content in residency practice compared with community practice: a study by the Kentucky Ambulatory Network. PMID- 17402319 TI - Ambulance transports from a state prison: a unique service staffed by prisoners. AB - OBJECTIVE: To describe an emergency medical service (EMS) staffed by convicted felons and to determine the nature of emergencies at a state penitentiary. METHODS: We retrospectively reviewed runs performed by the prison EMS for 2003. For each run, we recorded emergency run or a non-emergency transport, location of the patient, whether or not the run was staffed with a nurse or physician, and whether or not an automated external defibrillator (AED) was used. We also determined the cost of operating the service. RESULTS: During 2003, the prison EMS had a total call volume of 477 transports. The AED was used 8 times with one patient surviving to hospital admission neurologically intact. There were 357 emergency runs and 120 nonemergency transports. The majority of emergency transports, 305 (85%), were for medical complaints. The actual cost of operating the service for 2003 was $35,421. There were no attempted escapes or injured providers. This study shows that the majority of emergency transports in our prison were for medical conditions. CONCLUSION: The use of state prisoners as EMS providers results in substantial cost savings and most transports were for medical emergencies. PMID- 17402320 TI - Insulin therapy of hyperglycemia in intensive care. AB - BACKGROUND: Hyperglycemia is common among patients admitted to intensive care units, and carries the risk for complications and prolonged ICU stay. With intensive insulin control of blood glucose, morbidity and mortality can be reduced. OBJECTIVES: To determine whether intensive or conventional insulin control of blood glucose in hyperglycemic ICU patients correlated with the prognosis. METHODS: Following admission to the ICU, hyperglycemic patients were randomly assigned to a group treated intensively with insulin targeting glucose at 110-140 mg/dl, or to a conventional insulin therapy group, where glucose, upon exceeding 200 mg/dl, was controlled at 140-200 mg/dl. Rates of morbidity and mortality, hypoglycemic episodes, and insulin dosage were compared. RESULTS: In the 41 patients treated intensively with insulin the glucose level was 142 +/- 14 mg/dl, as compared to 174+/-20 mg/dl in the 48 patients on conventional insulin treatment. Both groups were similar in age, acute physiology and chronic health evaluation score. Morbidity was also similar, except for increased vascular damage in the conventional treatment group and slightly shorter ICU stay in the intensive therapy group. Both groups had similar in-ICU, in-hospital, and 28 day mortalities, and similar rates of hypoglycemic episodes. The daily dosage of insulin was significantly higher with the conventional treatment (P= 0.004). CONCLUSIONS: Intensive insulin treatment did not affect the mortality or morbidity rates in ICU patients. The increased insulin dosage of conventional insulin treatment was attributable to the group's higher prevalence of diabetes. Future studies should address this bias and determine the optimal glucose target. PMID- 17402321 TI - Plasmid-based gene transfer for treatment of erectile dysfunction and overactive bladder: results of a phase I trial. AB - BACKGROUND: Ion Channel Innovations has developed a gene transfer product, hMaxi K, and has begun clinical trials to investigate the effect of increased expression of Maxi-K channels in the smooth muscle of the penis or bladder in patients with erectile dysfunction and those with overactive bladder. The primary function of K channels is to modulate Ca++ influx through Ca-channels (i.e., L type, voltage-dependent). The amount of Ca++ that enters the cell through these channels is a major determinant of the free intracellular calcium levels inside the smooth muscle cell, which in turn determines the degree of smooth muscle cell contraction. Increased Maxi-K channel activity is associated with smooth muscle cell relaxation, resulting in, for example, penile erection and detrussor muscle relaxation. A phase I clinical trial that used hMaxi-K has been completed and a similar trial to assess safety of the transfer for overactive bladder is about to begin. OBJECTIVES: To assess the safety and tolerability of escalating hMaxi-K doses by clinical evaluations and laboratory tests, and to measure efficacy objectives by means of the International Index of Erectile Function scale. METHODS: In the erectile dysfunction trial 11 patients with moderate to severe erectile dysfunction were given a single-dose corpus cavernosum injection of hMaxi-K, a "naked" DNA plasmid carrying the human cDNA encoding for the gene for the alpha, or pore-forming, subunit of the human smooth muscle Maxi-K channel, hSlo. Three patients each were given 500, 1000, and 5000 pg and two patients were given 7500 microg doses of hMaxi-K and followed for 24 weeks. Patient responses were validated by partner responses. RESULTS: There were no serious adverse events and no dose-related adverse events attributed to gene transfer for any patient at any dose or study visit. No clinically significant changes from baseline were seen in physical evaluations (general and genitourinary), hematology, chemistry and hormone analyses, or in cardiac events evaluated by repeated electrocardiograms. Importantly, no plasmid was detected in the semen of patients at any time after the injections. Patients given the two highest doses of hMaxi-K had apparent sustained improvements in erectile function as indicated by improved IIEF-EF domain scores over the length of the study. One patient given 5000 microg and one given 7500 microg reported EF category improvements that were highly clinically significant and were also maintained through the 24 weeks of study. CONCLUSIONS: Efficacy conclusions cannot be drawn from results of a phase 1 trial with no control group. However, the promising primary safety outcomes of the study and preliminary indications of effectiveness provide evidence that hMaxi-K gene transfer is a viable approach to the treatment of erectile dysfunction and other smooth muscle diseases with targeted access. PMID- 17402322 TI - Comparison of the quality of life after minilaparotomy cholecystectomy versus laparoscopic cholecystectomy: a prospective randomized study. AB - BACKGROUND: Earlier studies comparing minilaparotomy cholecystectomy with laparoscopic cholecystectomy did not find significant differences between the MC and the LC groups in operating times and patients' recovery. OBJECTIVES: To compare the postoperative quality of life between the MC and LC groups. METHODS: The 157 patients with uncomplicated symptomatic gallstones, confirmed by ultrasound, were randomized to two groups: 85 for MC and 72 for LC. The study was prospective and randomized but not blinded or consecutive. The study groups were similar in patients' age, gender, body mass index, American Association of Anesthesiology physical fitness classification, and the operating surgeon. Patients were reevaluated 4 weeks after operation using the RAND-36 quality of life questionnaire. RESULTS: The RAND-36 questionnaire did not identify statistically significant differences between the study groups in general health perceptions, physical functioning, emotional well-being, social functioning, energy, bodily pain, and role functioning/emotional score. Only the role functioning/physical score was slightly higher in the LC group (P= 0.038). CONCLUSIONS: The results of this study showed that the MC procedure is a good alternative to the LC procedure, when postoperative quality of life is measured. PMID- 17402323 TI - Trends in specific morbidity prevalence in male adolescents in Israel over a 50 year period and the impact of recent immigration. AB - BACKGROUND: Most Israeli males aged 16-17 undergo a thorough medical examination prior to recruitment into the army. During the last 50 years, extensive data have been gathered enabling a study of time trends in the prevalence of common diseases in this age group. OBJECTIVES: To examine the current prevalence of common diseases, compare the results with those of previous cohorts, and assess the influence of the massive immigration during the 1990s. METHODS: The health examination at the recruitment centers includes a medical history, complete physical examination, and review of medical documentation provided by the family physician. If needed, additional tests and referral to specialists are ordered. The prevalence of selected diseases and severity was drawn from the computerized database of the classification board. Two cohorts, 1992-94 and 2003-04, were examined and compared with three previous cohort studies in 1957-61, 1977-78 and 1982-84. Data were stratified according to origin and country of birth. RESULTS: The prevalence of asthma increased dramatically during the years from 10.2 per 1000 examinees in 1957-61 to 111.6 per 1000 examinees in 2003-04. The prevalence of tuberculosis declined and then increased from 0.6 per 1000 adolescents in 1982 84 to 2.4 per 1000 adolescents in 2003-04. The prevalence of type 1 diabetes mellitus increased from 0.2 cases per 1000 examinees in 1957-61 to 0.8 cases in 1977-78 and 1982-84 and 0.9 cases per 1000 examinees in 2003-04. The prevalence of severe heart defects and severe epilepsy declined in the last 20 years (1.4 and 1.7 cases per 1000 examinees in the 1982-84 cohort to 0.4 and 0.3 cases per 1000 examinees in the 2003-4 cohort respectively). The patterns of disease prevalence were different for immigrants: tuberculosis was more common while asthma and allergic rhinitis were less prevalent. CONCLUSIONS: The prevalence of common diseases among adolescents in Israel has changed over the last 50 years. There is a different pattern for immigrants and for those born in Israel. PMID- 17402324 TI - Hepatocellular carcinoma: MRI and CT examination. AB - Hepatocellular carcinoma will continue to be one of the most common malignancies worldwide. Improved survival occurs following resection or liver transplantation. The appropriate pre-operative stratification and staging of these patients is essential. CT and MRI will undoubtedly continue to play a major role in the detection and diagnosis of HCC. These imaging techniques should be optimized for the evaluation of suspected HCC. The radiology report from the CT or MRI examination should include a comprehensive review of key diagnostic information for appropriate staging. This includes lesion size and number. Also to be noted are segmental and vascular involvement, regional and distant adenopathy as well as metastases, and finally, the presence of ascites, varices and cirrhosis. PMID- 17402325 TI - Human papillomavirus vaccine: the beginning of the end for cervical cancer. AB - The human papillomavirus family of viruses causes a variety of benign, premalignant and malignant lesions in men and women. All cervical cancers are caused by HPV. It is the leading cause of death from cancer in women in developing countries; every year some 493,000 women develop cervical cancer and 230,000 die every year from this disease. The vaccine against HPV includes virus like particles, composed of the major viral capsid protein of HPV without the carcinogenic genetic core. Large-scale studies have shown that the vaccine is tolerated well, leads to high antibody levels in both men and women, and prevents chronic HPV infection and its associated diseases. To achieve effective coverage the vaccine should be given prior to sexual debut. Introduction of the vaccine into specific countries, particularly Israel, should take into account the local incidence of cervical cancer as well as the increasing incidence of precancerous cervical lesions and genital warts, which reduce quality of life and are associated with considerable costs. PMID- 17402326 TI - Frontiers of stem cell transplantation monitoring: capturing graft dynamics through routine longitudinal chimerism analysis. AB - Quantitative chimerism testing has become an indispensable tool for following the course and success of allogeneic hematopoietic stem cell transplants. In this paper, we describe the current laboratory approach to quantitative chimerism testing based on an analysis of short tandem repeats, and explain why performing this analysis longitudinally is important and feasible. Longitudinal analysis focuses on relative changes appearing in the course of sequential samples, and as such exploits the ultimate potential of this intrinsically semi-quantitative platform. Such an analysis is more informative than single static values, less likely to be confused with platform artifacts, and is individualized to the particular patient. It is particularly useful with non-myeloablative conditioning, where mixed chimerism is common. When longitudinal chimerism analysis is performed on lineage-specific subpopulations, the sensitivity, specificity and mechanistic implications of the data are augmented. Importantly, longitudinal monitoring is a routinely feasible laboratory option because multiplex STR-PCR kits are available commercially, and modern software can be used to perform computation, reliability testing, and longitudinal tracking in a rapid, easy to use format. The ChimerTrack application, a shareware program developed in our laboratory for this purpose, produces a report that automatically summarizes and illustrates the quantitative temporal course of the patient's chimeric status. Such a longitudinal perspective enhances the value of quantitative chimerism monitoring for decisions regarding immunomodulatory post transplant therapy. This information also provides unique insights into the biological dynamics of engraftment underlying the fluctuations in the temporal course of a patient's chimeric status. PMID- 17402327 TI - Perianal Crohn's disease. AB - Perianal Crohn's disease refers to the involvement of the anal region in this chronic inflammatory bowel disease. It most commonly presents with the formation of perianal abscesses and fistulas, although other forms of presentations such as fissures and skin tags may also be present. Perianal activity often parallels abdominal disease activity, but may occasionally be the primary site of active disease, and significantly compromises the quality of life in affected patients. The primary treatment of patients with perianal Crohn's disease combines medical and surgical management with the aim of improving quality of life and alleviating suffering. A multidisciplinary approach involving the patient, surgeon, gastroenterologist, radiologist, pathologist, nutritionist, and other specialists makes the successful treatment of PCD possible. This paper reviews the management of patients with perianal Crohn's disease, focusing on contemporary medical and surgical treatments such as infliximab, endorectal advancement flap, instillation of fibrin glue, and the potential use of extracellular matrix plugs. PMID- 17402328 TI - Integration of complementary and alternative medicine services in the hospital setting in Israel. PMID- 17402329 TI - A rare cause of right upper quadrant pain. PMID- 17402330 TI - An alternative treatment for Parkinson's disease. PMID- 17402331 TI - Spontaneous bacterial peritonitis caused by Haemophilus parainfluenzae. PMID- 17402332 TI - Cardiopulmonary resuscitation in the frail elderly: clinical, ethical and halakhic issues. AB - Cardiopulmonary resuscitation is an emotion-ridden issue that often leads to conflicts when crucial decisions have to be made. The purported benefits of this 40 year old procedure in the frail elderly have been scrutinized, establishing its lack of efficacy. A review of the medical, ethics and halakhic literature on the potential merits of CPR in the frail elderly revealed that in secular medical practice, CPR is often routinely provided to elderly frail individuals for whom its clinical benefit is questionable. For patients suffering from dementia, surrogates are usually responsible for decision making, which complicates the process. With such poor clinical outcomes, the halakhic interpretation of what steps should be taken, and currently are, may not be valid and CPR may be applied too frequently. When clinical ambiguity is combined with strong cultural and religious influences, an acceptable CPR/DNR (Do Not Resuscitate) approach to cardiac arrest can be daunting. A clinically responsible, ethically sound and religiously sensitive approach to CPR requires a deep understanding of the factors involved in decision making. It seems timely for the halakhic interpretation of the duty to provide CPR in the frail elderly to be reevaluated. Perhaps a more humane and halakhically sound approach might be reached by stringently limiting CPR to clinically unusual circumstances rather than the common practice of providing frail Jewish elders with CPR in the absence of a DNR order. PMID- 17402333 TI - Prevention in Halakhah. AB - Preventive medicine is taking an increasingly central place in modern clinical practice, at least in primary care. What, if anything, does the Jewish rabbinic tradition have to say about keeping healthy? The delayed response of contemporary rabbis to the dangers of smoking, in particular, raises questions about the underlying principles that Halakhah employs to approach health promotion. As is often the case in Halakhah, we may detect different streams of thought in the classical sources, which may be felt in the way contemporary issues are handled. Three approaches will be discussed. First, Maimonides, famous for the practical preventive approach in his medical writings, makes his philosophy clear both in his halakhic works and in his Guide for the Perplexed. For him, a healthy body is a prerequisite for a healthy soul. We must be free of physical suffering in order to be able to do the work of perfecting our souls. Second, the view that health is the reward for goodness and illness a punishment for sin as expounded or implied in the writings of Nahmanides, and of Ibn Ezra: the way to good health is to lead a good life. Third, an early midrashic source picked up again much later by Rabbi Israel Meir Kagan (the Hafetz Hayim) gives the argument from custodianship--since the body is divine property we have a duty to look after it well. So for Maimonides there is a prior duty to keep healthy, while for Nahmanides the prior requirement is to repent of sin. For the Hafetz Hayim, keeping the body healthy is an independent duty in its own right. These then are the differences in basic approach that may affect the emphases that different rabbis today place on health maintenance and promotion. PMID- 17402334 TI - Report on the Anglo-Israeli workshop on the genetic risk of breast cancer. PMID- 17402335 TI - Light from the flames of Hell: remembrance and lessons of the Holocaust for today's medical profession. PMID- 17402336 TI - Holocaust and medicine--a medical education agenda. PMID- 17402337 TI - The Nazis and medical ethics: the context. PMID- 17402338 TI - Justice in Nuremberg: the doctors' trial--60 years later. A reminder. PMID- 17402339 TI - The aging of Holocaust survivors: myth and reality concerning suicide. AB - The association between the Holocaust experience and suicide has rarely been studied systematically. The dearth of data in this area of old-age psychiatry does not necessarily imply that Holocaust survivors are immune from suicide. Recent work on the aging of survivors seems to suggest that as a group they are at high risk for self-harm. Published reports on suicide and the Holocaust identified by means of a MEDLINE literature search were reviewed. A similar search was performed on the Internet using the Google search engine. Thirteen studies were uncovered, 9 of which addressed the association of suicide and the Holocaust experience and 4 focused on suicide in the concentration camps during the genocide. Eleven of the 15 studies explicitly reported on the association of suicide, suicidal ideation or death by suicide with the Holocaust experience, or reported findings suggesting such an association. The Internet search yielded three sites clearly describing increased suicide rates in the concentration camps. An increased rate of suicidal ideation and suicide attempts among the elderly who were exposed to the Holocaust experience is confirmed. There is a need for further study, intervention and resource allocation among the growing numbers of elderly persons who suffered traumatic events in earlier phases of their lives. This is especially critical for Holocaust survivors. PMID- 17402340 TI - The limits of starvation--personal experience. PMID- 17402341 TI - Doctors, pregnancy, childbirth and abortion during the Third Reich. AB - This paper does not attempt to deal with the legitimate ethical or moral debate on abortion. Utilizing abortion as a subject I will show how science and medicine in general, and abortion in particular, were used as weapons of mass destruction by Nazi physicians in their zeal to comply with the political climate of the time. Nazi policy on abortion and childbirth was just one of the methods devised and designed to ensure the extermination of those whom the Nazis deemed had "lives not worth living." Physicians implemented these policies, not with the fate of their patients in mind, but rather in the name of the "state." When discussing pregnancy, abortion and childbirth during the Holocaust it is imperative to include an essay of how these issues affected the Jewish prisoner doctors in the ghettos and camps. Nazi policy dictated their actions too. From an extensive search of their testimonies, I conclude that for these doctors ethical discourse comprised a fundamental component of their functioning. I do not propose to judge them in any way and one should not, in my opinion, argue whether their behavior was or was not morally acceptable under such duress; nevertheless, unlike their Nazi counterparts, a key theme in their testimonies was to "keep their medical values." PMID- 17402342 TI - Eponyms and the Nazi era: time to remember and time for change. AB - Eponyms are titles of medical disorders named for individuals who originally described the condition. They also help us remember and identify the disorder. Medicine is replete with them, and changing them or eradicating them, for whatever reason, is not simple. But when there is a moral issue involved - for example, research conducted under overwhelming unethical conditions - we believe it wrong to perpetuate and thus "rew ard" the memory of the individual for whom the disorder is named. The name of a syndrome should thus be discontinued if described by an individual whose research used extreme measures or who was involved in atrocities against humanity. Ethical considerations should be introduced into medical nosology just as they exist in patient care and research. This article details a group of notable eponyms, the names of which are associated with overt crimes of the medical community during the Nazi era, and provides alternative medical nomenclature. In addition, examples are provided of eponyms named after Nazi era victims, eponyms of those who protested such injustices, and eponyms of those who had to flee discrimination and death. These should be remembered and even strengthened, as opposed to those of the perpetrators, which should be obliterated. Since the greatest accolade a physician can earn is praise from his colleagues as expressed in an eponym entrenched in one's name, the medical profession should remove any honor given to physicians involved in crimes to humanity. PMID- 17402343 TI - Ole Secher: a true hero of anesthesiology. PMID- 17402344 TI - Self-care programme to prevent leprosy-related problems in a leprosy colony in Champa, Chattisgarh. AB - A self-care programme aimed at preventing leprosy-related physical disabilities in a leprosy colony in Champa, Chattisgarh, India, is described. Once the initial resistance was overcome through persistent and caring attitudes, the residents accepted the challenges for self-care. The outcome at the end of one year showed significant decline (41%) in ulcer rates, significant use of MCR footwear (43%), and significant proportion (over 90%) of patients and their families practising and helping in self-care activities. PMID- 17402345 TI - Evaluation of a new fixed duration (12 weeks) multi-drug regimen of bactericidal drugs in multibacillary leprosy. AB - Leprosy still remains a public health problem mainly in Asia, Africa and South America. The WHO Expert Committee on Leprosy recommended, in 1997, the simplified treatment of leprosy for multibacillary (MB) cases, by reducing the duration of treatment from 24 to 12 months. From the operational point of view even this reduced duration is still long and monthly supervised drug administration may not always be practical in those areas where the accessibility is difficult and health infrastructure weak. The present study was carried out to compare the safety and efficacy of a new fixed duration regimen consisting of four bactericidal drugs with WHO/MDT (MB). METHODS: Thirty adult patients were randomly allocated to two groups. Group 1 (18 patients) received a new regimen of daily rifampicin 600 mg, sparfloxacin 200 mg, clarithromycin 500 mg and minocycline 100 mg for 12 weeks. Group 2 (12 patients) received WHO/MDT (MB) for 12 months. A detailed clinical evaluation and laboratory investigations, BI and MI were done at the baseline, every 4 weeks for 12 weeks, and thereafter every 8 weeks till 48 weeks. Skin biopsies were taken and chest X-rays were done at 0, 12 and 48 weeks. RESULTS: At 48 weeks, the net percentage clinical improvement in group 1 was 73.92% and in group 2 it was 66.66%. The net percentage reduction (NPR) in BI in group 1 was 19.17% and in group 2 it was 18.87% (p = 0.09). NPR in MI in both groups was 100% by 8 weeks, and somewhat faster in group 1. In group 1, 8 patients had mild gastrointestinal side-effect, and 16 had minocycline induced hyperpigmentation. Three patients in group 1 developed type I reversal reactions. CONCLUSION: The regimen containing daily doses of rifampicin 600 mg, sparfloxacin 200 mg, minocycline 100 mg and clarithromycin 500 mg for 12 weeks was found to be an acceptable, effective and safe alternative regimen for MB leprosy with an additional operational advantage of shorter duration of treatment. PMID- 17402346 TI - Comparative profile of new leprosy cases coming to a referral institute in pre- and post-integration periods. AB - This article presents findings from a retrospective analysis of 3031 new leprosy cases attending the Out-patient Department of the Regional Leprosy Training and Research Institute, Raipur, Chhattisgarh, during the period January 2000-December 2005. The analysis was carried out in order to compare the profiles of new leprosy cases between pre- and post-integration periods. The cut-off date, in order to distinguish between the two groups, was determined as 31st December 2002. A comparison was made between the two groups on identified host- and disease-related variables and its significance was tested using suitable statistical tests. The results showed that there was more than 50% increase in the number of new leprosy cases coming to the OPD of the RLTRI during the post integration period. The mean age at registration showed an increase of 0.9 years, while the proportion of child and female cases showed a decrease of 7.7% and 4.3% respectively during the same period. Significant decline was also recorded in the proportion of grade II deformity cases (11.8%) and cases with type I lepra reaction (11.4%) during the post-integration period. The study emphasizes the need for developing an effective referral mechanism and proper screening of cases at the PHC level for better utilization of referral institutions in a need-based manner. PMID- 17402347 TI - Prevention-of-impairment-and-disabilities activities in leprosy after integration: role for physical medicine and rehabilitation personnel. AB - Referral options for specialist care for prevention of impairment and disabilities are imperative in order to make an integrated leprosy control system work. This requires an understanding of the disease, in addition to the special skills for managing specific disabilities. Physical medicine and rehabilitation (PMR) personnel are better equipped to handle leprosy-related disabilities. They are well versed with biomechanical aspects of deformities, and are competent to provide splints, orthoses, etc. to the needy persons, and they can assess sensory motor functions and deformities. If PMR personnel can be trained in deformity correction they can become valuable resource persons for secondary and tertiary care of leprosy-affected persons. PMR persons, therefore, have the opportunity to volunteer themselves for this job to fill the void created by the fading out of leprosy surgeons. They will also have to bear additional responsibility to train general health care workers so as to empower them to look after the needs of those disabled by leprosy, many of whom will continue to be available for a number of years to come. PMID- 17402348 TI - Dapsone syndrome with leukemoid reaction. PMID- 17402349 TI - Cutaneous sarcoidosis masquerading as lepromatous leprosy. PMID- 17402350 TI - Co-existent leprosy and psoriasis: an unusual presentation. PMID- 17402351 TI - [Coronary allograft vasculopathy: pathophysiological interaction between the immune system, infections and metabolic syndrome]. AB - Cardiac allograft vasculopathy is still the main cause of long-term graft loss after heart transplantation. Indeed, recent advances in immunosuppression management led to a significant improvement in short-term survival, while long term death rate did not change significantly in the last 20 years. In this paper, we will review the latest advances in the understanding of this peculiar form of atherosclerosis, focusing on the mechanisms that can be potentially targeted by specific therapeutic interventions. PMID- 17402352 TI - [Disease management system in patients with chronic heart failure]. AB - Healthcare managers are more and more interested in the role of general practitioners (GP) in the treatment of cardiovascular diseases. Continuing adjustments of the health organization are the old/new challenge in improving patient care. The European Society of Cardiology guidelines recommend a disease management program for heart failure (HF); moreover, observational studies and randomized controlled trials have reported better patient outcomes if patients are in charge of cardiologists rather than GPs or other physicians. Patients with chronic HF are often very old and affected by multiple comorbid conditions, by themselves associated with high rates of morbidity and mortality. Furthermore, too many patients receive neither a correct diagnosis nor treatment until advanced disease occurs. New treatment approaches, some of them requiring the expertise of well-trained cardiologists, are ongoing to improve the clinical outcomes. The optimal management of patients with HF needs teamwork, i.e. GPs, cardiologists, nurses and caregivers, since a multidisciplinary program, only, can embody the best answer for outpatients with chronic HF. Currently, the Cardiovascular Center in Trieste is performing an experimental trial, so far never attempted before, in treating patients with chronic HF using a thorough approach with the full involvement of local cardiologists, GPs and nurses. Such approach is, at the same time, as well a challenge as an opportunity: a challenge because conventional clinical habits must be changed; an opportunity because patients can benefit from a proper whole care-group, aimed at prolonging life and reducing morbidity and symptoms. PMID- 17402353 TI - [Pulsed tissue Doppler illustrated to a resident in cardiology]. AB - Tissue Doppler is an ultrasound tool representing a useful completion of standard Doppler echocardiographic examination, The placement of pulsed tissue Doppler sample volume at the level of the different sites of the mitral annulus allows to obtain information on left ventricular global longitudinal (systolic and diastolic) function, whereas the sampling of tricuspid lateral annulus gives similar information on the right ventricular chamber. Myocardial early diastolic velocity (Em) is a marker of ventricular diastolic relaxation and the ratio obtainable between transmitral (or transtricuspid) E velocity and annular Em (E/Em ratio) is an accurate index of the level of filling pressure of the assessed ventricular chamber. Myocardial systolic velocity (Sm) as well as the frequency of acceleration of presystolic velocity, occurring during isovolumic contraction time, are indexes of systolic performance and their reduction is evident in several cardiac pathologies. Tissue Doppler-derived systolic and diastolic indexes have a recognized prognostic value in heart failure and after acute myocardial infarction. In these conditions they are able to predict both left ventricular remodeling and mortality. PMID- 17402354 TI - [Temporal trends in pharmacological therapy in the IN-CHF registry from 1995 to 2005]. AB - In the last years the treatment of heart failure has radically changed due to the results of multicenter clinical trials. The antagonism of neurohormonal systems has proved to be the only strategy, which favorably modifies the prognosis of the patients with heart failure. Particularly, the effectiveness of angiotensin converting enzyme (ACE)-inhibitors and beta-blockers has been proven in patients with heart failure and left ventricular dysfunction; more recently, angiotensin receptor blockers (ARBs) and aldosterone blockers have shown to improve the outcome in heart failure. The public health im portance of translating this body of evidence of research into clinical practice is paramount. We used data from the IN-CHF registry to examine changes in the use of pharmacological treatment from 1995 to 2005. The proportion of patients receiving an ACE-inhibitor was slightly higher in the 1995-2000 than after but this difference is not statistically significant. The use of "recommended therapies" as beta-blockers, aldosterone antagonists and ARBs increased significantly in these 10 years. The use of beta-blockers also increased significantly among elderly patients and patients in advanced NYHA class. Patients treated with the combination of ACE inhibitors and ARBs are very few (1.1%), even if this association is now recommended by the European Society of Cardiology guidelines. The same analysis repeated after the CHARM and Val-HeFT publication shows that this association is used in 2.0% of the patients. The proportion of patients treated with a beta blocker plus ACE-inhibitors plus ARB is only 0.8%. Use of digitalis and calcium channel blockers fell continuously from 1995 whereas use of diuretics and anticoagulants remained relatively constant. Statins are becoming widely used in this population (from 5 to 20%) even if the information on the effect of these drugs in heart failure is still incomplete. PMID- 17402355 TI - [Disease management for heart failure patients: role of wireless technologies for telemedicine. The ICAROS project]. AB - Healthcare costs for heart failure are increasing. The need for a better care, however, has to be matched with a policy of cost containment. A way to improve the cost-effectiveness of heart failure care is the disease management approach, in which therapy, education and follow-up are tailored for each patient by a multidisciplinary team. Such a complex intervention can be facilitated by the use of telemedicine, which allows the remote control of considerable amounts of clinical data. In Italy, a few studies with telemedicine have been reported. A recent development in this field is represented by the ICAROS project (Integrated Care vs Conventional Intervention in Cardiac Failure Patients: Randomized Open Label Study), whose aim is to improve the clinical and psychological care of heart failure patients employing advanced wireless telecommunication technology. So far, we randomized 60 patients: 30 in usual ambulatory care, 30 in an intensive treatment group. The latter patients were instructed to use a portable computer to get in touch daily with the heart failure clinic and receive feedback instruction for the management of drug therapy and daily problems. At the first year of follow-up, the treatment group showed good compliance to drug prescriptions, and could easily handle the portable computer. The preliminary results of this ongoing study support the feasibility and appropriateness of new technologies for the management of heart failure, even in elderly patients in whom a limited expertise with these appliances could have been anticipated. PMID- 17402356 TI - [Heterozonal electrocardiographic changes in ST-elevation myocardial infarction]. AB - BACKGROUND: It has been observed that in patients with ST-elevation myocardial infarction (STEMI) the presence of ST-segment depression (ST) in heterozonal electrocardiographic leads (remote STI) worsens the patient's prognosis. The aim of this study was to observe in an unselected population with a first STEMI the incidence of remote STI and the risk factors related to this condition. METHODS: We evaluated retrospectively 350 patients with a first STEMI; we excluded from our analysis 139 patients because no data about their coronary anatomy was available. ST-segment depression in the heterozonal myocardium was considered significant if > 0.1 mV at 60 ms from the J point, in at least two electrocardiographic leads. RESULTS: Patients were classified according to the presence (group 1, 117 patients) or absence (group II, 94 patients) of remote ST. The two groups did not differ for age, sex and coronary anatomy. In group I we found more heterozonal wall motion abnormalities than group II (32 vs. 18%, p = 0.018). In this group there was a higher incidence of smokers (56 vs. 33%, p = 0.025) and less patients were treated with statins when the STEMI occurred (6 vs. 14%, p = 0.047). Patients with remote ST had higher total cholesterol (214.6 +/- 48.9 vs. 192.3 +/- 29.8 mg/dl, p < 0.001) and low-density lipoprotein cholesterol (138.7 +/- 40.7 vs. 123.2 +/- 22.9 mg/dl, p < 0.0001) levels. Conclusions. In patients with STEMI the presence of remote ST is rather frequent, and seems to indicate a real heterozonal ischemia, independently of an epicardial coronary stenosis of the non-infarct-related artery. Remote ST. is associated with a higher incidence of risk factors related to microcirculatory dysfunction, such as cigarette smoking, a worse lipid profile and less protective factors, such as the use of statins prior to acute myocardial infarction. PMID- 17402357 TI - [Anomalous origin of the right coronary artery from the left sinus of Valsalva: case report and literature review]. AB - We describe the case of a patient in whom evaluation of effort angina revealed a tight stenosis of a right coronary artery anomalously arising from the left sinus of Valsalva, which was successfully treated by stent implantation. The abnormal origin of the right coronary artery from the left aortic sinus coursing between the aorta and the pulmonary trunk is a rare congenital anomaly. It may remain asymptomatic, but can also cause major cardiac events, even in the absence of coronary atherosclerosis. We discuss the clinical importance of this anomaly and review the literature concerning current views and therapy. PMID- 17402358 TI - [Heart failure and cardiomyopathies: a case report]. AB - Isolated noncompaction of left ventricular myocardium is a rare congenital heart disease, characterized by an excessive prominence of trabecular meshwork, spaced out by deep intertrabecular recesses, consequent to the arrest of the normal myocardial embryogenesis. Although there are numerous descriptions, the pathophysiological effects of the structural alterations, like the clinical spectrum and the evolution of the disease, are not fully clarified. In this paper we evaluated the natural history of the disease, the family incidence and the alterations of the systolic and diastolic function. An interesting case report is described concerning a patient affected by noncompaction and atrial fibrillation. PMID- 17402359 TI - [Response to the open letter of the President-Elect of the European Society of Cardiology Roberto Ferrari, published in Il Giornale Italiano di Cardiologia 1, 2007]. PMID- 17402360 TI - [Response to the open letter of the President-Elect of the European Society of Cardiology Roberto Ferrari, published in Il Giornale Italiano di Cardiologia 1, 2007]. PMID- 17402361 TI - An introduction to the National Nursing Assistant Survey. AB - OBJECTIVES: This report provides an introduction and overview of the National Nursing Assistant Survey (NNAS),the first national probability survey of nursing assistants working in nursing homes. The NNAS was designed to provide national estimates and to allow for separate estimates to be calculated for nursing assistants by geographic location of the agency and for workers by tenure at the sampled facility. This report includes a description of relevant research that led to federal interest in sponsoring the NNAS, types of data collected, methodology, linkage between the NNAS and the 2004 National Nursing Home Survey (NNHS), advantages of combining establishment and worker surveys, and potential uses of the data. METHODS: The NNAS was conducted as a supplement to the 2004 National Nursing Home Survey. The design was a stratified, multistage probability survey. Nursing facilities were sampled and then nursing assistants were sampled within the facilities. Telephone interviews were conducted with nursing assistants using Computer-Assisted Telephone Interviews (CATI). The survey instrument consisted of sections on recruitment, training and licensure, job history, family life, management and supervision, client relations, organizational commitment and job satisfaction, workplace environment, work related injuries, and demographics. RESULTS AND CONCLUSIONS: A total of 3,017 interviews were completed from September 2004 to February 2005. The overall response rate was 53.4 percent. A public-use data file has been released that contains the interview responses and sampling weights. The file also includes ownership, bed size, and geographic location of the facility where the nursing assistant was sampled. Estimates based on the sampling weights can be used to produce national estimates. PMID- 17402362 TI - High prevalence of antibodies to platelet factor 4 heparin in patients with antiphospholipid antibodies in absence of heparin-induced thrombocytopenia. AB - Seventy-two patients with antiphospholipid antibodies (aPL), with or without antiphospholipid syndrome (APS), were studied for detection of heparin-PF4 induced antibodies (HPIA) using a commercial kit (Asserachrom HPIA) PF4-dependant enzyme-linked immunoassay (ELISA) test. None of the patients had a medical history of heparin induced thrombocytopenia (HIT). Eleven percent of patients were positive for HPIA. Plasma from 40 of the 72 patients (seven positive and 33 negative), was also tested with the other available HPIA ELISA (GTI) kit. Five patients were positive with both ELISA kits, two were highly positive only with Asserachrom HPIA and four only with GTI. None of the positive patients had severe thrombocytopenia. Two patients have never received heparin treatment. No relationship was found between HPIA presence and patients' age, sex, aPL levels or presence of lupus anticoagulant. No significant difference in HPIA presence was observed in patients with primary APS, secondary APS or aPL without APS. We found a poor correlation between the two commercial ELISA showing that, on the same blood sample, a patient could be highly positive with one technique and negative with the other. The PF4-dependant enzyme-linked immunoassay, which is often the first test used for the diagnosis of HIT, should be interpreted cautiously in patients with aPL since there is a danger of overdiagnosis and overtreatment. PMID- 17402363 TI - Impaired endothelial function in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) patients suffer from excess cardiac deaths due to accelerated atherosclerosis. Endothelial dysfunction is a marker of early atherosclerosis. We tested the hypothesis that SLE patients have impaired endothelial function and assessed the relationship between endothelial function and clinical outcome over the subsequent five years. Thirty-six female SLE patients were compared with 22 healthy age and sex matched controls. Endothelial dependent vasodilatation (EDD) was assessed at the brachial artery in response to shear stress. Endothelium-independent dilatation induced by glyceryl trinitrate was also measured. Patients were followed for up to five years and the development of damage in the cardiovascular and other systems recorded. SLE patients showed significantly impaired endothelial function (median EDD 5.6%, IQR 3.1-7.2%) compared with healthy controls (median EDD 8.0%, IQR 6.3-9.3%; P = 0.001). Endothelium independent dilatation did not differ between the two groups. Endothelial function was significantly worse in postmenopausal compared with premenopausal women (median EDD 6.6%, IQR 3.9-7.8% versus 3.1%, IQR 2.6-5.1%; P = 0.016). Total cholesterol was inversely correlated with endothelial function in SLE patients (Spearman correlation r = -0.422, P = 0.025). There was no relationship between endothelial function and the development of damage in any organ system, including the cardiovascular system during patient follow-up. Patients with SLE have impaired endothelial Lupus (2007) 16, 84-88. PMID- 17402364 TI - Chloroquine treatment reduces the number of cutaneous HLA-DR+ and CD1a+ cells in patients with systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a chronic autoimmune disease that can be exacerbated by exposure to ultraviolet radiation (UVR). The number and phenotype of antigen presenting cells in the skin play a role in cutaneous immune response generation. Although antimalarials are widely used in SLE treatment, their mode of action is not completely elucidated. The aim of our study was to determine the effect of chloroquine treatment on HLA-DR+ and CD1a+ cell number in locally irradiated (three minimal erythema doses of UVB) and normal appearing skin in SLE patients and healthy subjects. A significantly higher number of HLA-DR+ and CD1a+ cells were found in both locations in SLE patients compared with controls. Following three months of daily chloroquine treatment (250 mg), the HLA-DR+ and CD1a+ cell counts were significantly reduced in both irradiated and unirradiated sites of SLE patients, although still higher than in controls. Chloroquine treatment reduces the number of antigen presenting cells in the skin of SLE patients, and this effect may explain the antimalarials beneficial immunoregulatory and anti-inflammatory properties. PMID- 17402365 TI - Overexpression of the antiapoptotic gene Bfl-1 in B cells from patients with familial systemic lupus erythematosus. AB - Genetic determinants taking part in the development of systemic lupus erythematosus (SLE) are complex and not fully characterized. Dysregulated expression of genes involved in the control of apoptosis has been previously suggested. We report here a consanguineous family with SLE manifestations in three siblings associated in one of them with severe lymphoproliferative features. Laboratory studies showed no defect in CD95-mediated cell death. Screening expression of Bcl-2 family genes that regulate mitochondrial apoptosis pathway showed an overexpression of the antiapoptotic Bfl-1 gene. Real time RT PCR analysis indicated that overexpression of Bfl-1 was restricted to B-cells, with normal expression in T-cells. Those results suggest that overexpression of Bfl-1 could result in impaired B-lymphocyte homeostasis and inappropriate immune response leading to autoimmune manifestations. PMID- 17402366 TI - Activation of the STAT1 signalling pathway in lupus nephritis in MRL/lpr mice. AB - Interferons (IFNs), type I (alpha/beta) and type II (gamma), comprise a family of multifunctional cytokines with antiviral, antiproliferative and immunomodulating properties. Both type I and type II IFNs have been heavily implicated in the pathogenesis of systemic lupus erythematosus (SLE). The biological effects of IFNs are mediated through the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in which both IFN-alpha/beta and IFN-gamma activate the transcription factor STAT1. However, little is known about the pathogenic significance of STAT1 in SLE. At this point, we examined the expression and activation of STAT1 in the kidney of MRL/lpr mice with lupus nephritis (LN) by immunohistochemistry, Western botting and real time quantitative RT-PCR. Increased levels of total STAT1 protein and its activated/phosphorylated form were detected in kidney samples from MRL/lpr mice with LN as compared to those from control mice. Phosphorylated STAT1 was predominantly detected in glomeruli cells. Gene expression of the STAT induced feedback inhibitors suppressor of cytokine signalling-1 (SOCS-1) and SOCS-3 was also enhanced in MRL/lpr mice. In MRL/lpr mesangial cells, both IFN-alpha and IFN gamma rapidly induced the phosphorylation of STAT in vitro. Our results demonstrate that expression and activation of STAT1 are significantly increased in murine lupus nephritis, and indicate that STAT1 signalling pathway may play an important role in the pathogenesis of kidney inflammation. PMID- 17402367 TI - Reduced placental growth and hCG secretion in vitro induced by antiphospholipid antibodies but not by anti-Ro or anti-La: studies on sera from women with SLE/PAPS. AB - Systemic lupus erythematosus (SLE) and primary anti-phospholipid syndrome (PAPS) are autoimmune diseases causing recurrent pregnancy loss. We hypothesized that anti-phospholipid antibodies (aPL), but not anti-Ro and anti-La antibodies, might have a role through direct placental damage. We cultured human placental explants in sera from women with SLE/PAPS with different antibodies. These sera were found to reduce placental growth and increase trophoblastic apoptosis. No effect was found on estradiol or progesterone secretion, but inhibition in betahCG secretion was detected. BetahCG was reduced in women with a history of recurrent pregnancy loss or thromboembolic events, and was also the most sensitive marker when examining the effects of specific antibodies. High titers of aPL were found to cause the largest reduction in betahCG. Anti-Ro and anti-La did not induce placental damage. A strong correlation was found between the rise in the number of different antibodies in the sera and the incidence of recurrent pregnancy loss, which was also accompanied by a decline in the betahCG levels. In conclusion, aPL, but not anti-Ro or anti-La, may cause placental damage in vitro. Thus betahCG levels might constitute a predictive marker for the risk of placental damage and pregnancy loss in women with SLE/PAPS. PMID- 17402368 TI - Abnormality of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus. AB - Bone marrow-derived mesenchymal stem cells (MSCs) are key components of the hematopoietic microenvironment and provide support to hematopoiesis and modulate immune system. Several studies suggest that SLE may be seen as stem cell disorders. However, it is unclear that whether MSCs from SLE patients are defective. So in this research, we studied the biological character of bone marrow derived MSCs in patients with SLE, focused on their phenotype (morphology and immunophenotype), karyotype, cytokines expression and hematopoietic support of MSCs. Our results showed that MSCs from SLE patients and normal controls can be successfully culture-expanded, but the MSCs from SLE grew more slowly than those of normal controls (P < 0.05). Cells from both groups were positive for CD29, CD44 and CD105, and negative for CD14, CD34, CD45 and HLA-DR. MSCs from SLE have a normal karyotype. Both groups express IL-6, IL7, IL-11, macrophage colony stimulating factor (M-CSF) and stem cell factor (SCF) at mRNA level. While IL-6 and IL-7 were down-regulated in MSCs from SLE patient (P < 0.05) at mRNA level. The MSCs from SLE patients and normal controls were infused into ICR (Tac: Icr: Ha strain) mice after high-dose chemotherapy, with no adverse events in either group. Recovery of white blood cells, hemoglobin and platelet was more rapid (P < 0.05) compared with the group without MSCs infusion. We conclude that MSCs in patient with SLE have abnormalities compared with those in normal control. MSCs in patient with SLE may play an important role in the SLE pathogenesis. PMID- 17402369 TI - Response to tetanus vaccination in infants exposed in utero to immunosuppressants for maternal autoimmune disorders. AB - Immunosuppressive drugs given during pregnancy to mothers suffering from a systemic autoimmune disease (AID) can cross the placenta, thus being potentially able to affect the offspring immune system. Aim of our study was to evaluate the in vivo immune function of a series of these newborns. Twenty-two babies born from mothers suffering from autoimmune diseases (AID) who had been taking immunosuppressive drugs during pregnancy were evaluated for their response to vaccination with C. Tetani toxoid. Six babies born from mothers receiving low dose aspirin only were used as controls. The immune response to C. Tetani vaccination was evaluated with an ELISA to detect circulating antibodies. Five children out of 28 (17%) did not achieve a protective titer of anti C. Tetani toxoid IgG. No clear relationship was found between specific drug exposure and antibody response. Our findings suggest that maternal immunosuppressive treatment given for a systemic AID can affect the response to an active immunization, without specificities for drug types used. PMID- 17402370 TI - Autoimmune pancreatitis as the initial presentation of systemic lupus erythematosus. AB - Pancreatitis is a relatively rare complication in systemic lupus erythematosus (SLE). Here we present a case of SLE associated with autoimmune pancreatitis. A 37-year old woman was admitted to our hospital because of fever, skin rash, proteinuria and abdominal pain. A diagnosis of SLE was made based on her clinical, laboratory and renal histological findings showing diffuse proliferative lupus nephritis. Elevated serum amylase, typical radiographic findings and selective increase in serum IgG4 all suggested that the patient also had autoimmune pancreatitis. Systemic administration of glucocorticoid successfully induced remission of pancreatitis and nephritis along with the reduction of IgG4. Autoimmune pancreatitis is a newly recognized type of pancreatitis, in which IgG4 immune response is thought to participate pathophysiologically. Although the disease has been observed to develop in patients having various connective tissue diseases, our report is the first to describe IgG4-related autoimmune pancreatitis in a patient with SLE. PMID- 17402371 TI - Acute gastrointestinal distress syndrome in patients with systemic lupus erythematosus. AB - Cases of mesenteric vasculitis in systemic lupus erythematosus (SLE) are well described, however, individual patient with recurrent mesenteric vasculopathy producing repeated episodes with each exacerbation similar in character and quality has not been reported previously in the literature. We describe two SLE patients whose condition was complicated by repeated stereotypic, CT confirmed, episodes of lupus enteritis characterized by dramatic intestinal wall edema. Moreover, each flare was accompanied by significant hypocomplementemia and was rapidly reversible suggesting an acute gastrointestinal distress syndrome (AGDS) as a result of leukoaggregation and a gut capillary leak syndrome. PMID- 17402372 TI - Outcome of a national Israeli cohort of pediatric systemic lupus erythematosus. AB - The aim of this study was to describe the clinical manifestations and outcomes of a national cohort of childhood systemic lupus erythematosus (cSLE). All cases of cSLE registered in the Israeli national registry of children with rheumatic diseases between 1987-2003 were examined for disease activity and damage by the SLE disease activity index (SLEDAI) and SLE collaborating clinics/American College of Rheumatology (SLICC/ACR) damage index. Demographic, clinical, laboratory and treatment factors were analysed for their effect on the outcome. One-hundred and two patients were identified, 81% females, with a mean age at diagnosis of 13.3 +/- 2.6 years. The mean SLEDAI score was 17.2 +/- 9.0 (range 2 60). Fifty four patients were followed for at least five years. The mean SLEDAI decreased to 7.6 +/- 6.3 (0-29) and the mean SLICC/ACR damage index was 0.7 +/- 1.6 (0-8). Five patients developed chronic renal failure. No patients died. No factors were found to be significantly associated with the outcome except the initial SLEDAI score. The five-year outcome of our national cSLE cohort was good; with relatively low activity and minimal damage in most patients. The initial SLEDAI predicted the development of late damage. PMID- 17402373 TI - Increased frequency of mannose-binding lectin promoter LX haplotype among Filipinos with systemic lupus erythematosus. PMID- 17402374 TI - The value of evidence-based continuing education. PMID- 17402375 TI - Diabetes mellitus update. PMID- 17402376 TI - Using podcasts for nursing education. PMID- 17402377 TI - Barriers to participation in continuing education activities among rural and remote nurses. AB - BACKGROUND: This article examines the barriers to participation in continuing education activities that are perceived by rural and remote registered nurses in Canada. METHODS: The data are drawn from a national survey that was part of a larger national project, "The Nature of Nursing Practice in Rural and Remote Canada." RESULTS: Perceived barriers to participation in continuing education activities include the isolation of rural nurses and time and financial constraints. Nurses who perceived barriers to participation were more likely to be middle-aged, unmarried, and working full-time than nurses who did not perceive barriers. They were also more likely to possess higher levels of nursing education and have children or dependents. The perception of barriers to participation was also associated with lower job and scheduling satisfaction. CONCLUSIONS: Rural and remote registered nurses have moderately high levels of participation in continuing education; however, participation and job satisfaction can be improved if some of the barriers identified are addressed. PMID- 17402378 TI - Nursing clinical faculty revisited: the benefits of developing staff nurses as clinical scholars. AB - One of the major contributing factors to the nursing shortage in Colorado and nationally is a scarcity of clinical faculty required to prepare new nurses. This article describes an innovative, collaborative project that purposefully prepares staff nurses to assume the role of clinical scholar. Although there are several models that have sought to make clinically expert nurses available as teaching faculty, the breadth and scope of the described project with its multiple partners is unique. In the first year of this grant, the identified goals were achieved. In addition, there have been significant serendipitous outcomes. PMID- 17402379 TI - Three key components to successfully completing a nursing doctoral program. AB - Using their personal experience, the authors identify three key components to successfully completing a nursing doctoral program. The first component is asking the right questions to determine the doctoral degree best suited for the potential student. The second component is having a strategic plan that includes aspects such as financial considerations (research costs and potential financial support) and the development of a strong support system (specifically study pairs). The third component is the development of a systematic approach to completing the research, writing the dissertation, and completing the oral requirements. Addressing these areas will help students balance the many demands of doctoral study. PMID- 17402380 TI - Risk factor outcome comparison between exercise-based cardiac rehabilitation, traditional care, and an educational workshop. AB - BACKGROUND: This study examined risk factor outcomes among patients who attended cardiac rehabilitation sessions, those who received traditional care, and those who attended Leap for Life workshops. METHODS: A non-equivalent, three-group design was used in this observational study. Baseline and 12-month measurements were collected for 217 participants. Analysis of covariance was performed to determine differences between groups on outcome variables. RESULTS: The only significant finding was in participants with an initial high-density lipoprotein value of less than 40. High-density lipoprotein levels increased more in the cardiac rehabilitation group than in the traditional care group (30.54 to 37.48 versus 30.17 to 33.67 [F= 4.577, p = .035]). CONCLUSIONS: Based on these findings, a strong case can be made for the transition to more individually intense and focused risk factor modification strategies for patients in cardiac rehabilitation programs. PMID- 17402381 TI - Images of loneliness: using art as an educational method in professional training. AB - BACKGROUND: An educational program using art was created to prepare healthcare professionals to work with older adults by deepening their knowledge of loneliness and promoting self-reflection. METHOD: Participants viewed art exploring themes of loneliness and discussed their perceptions. Semi-structured feedback questionnaires were used to evaluate evidence of learning. RESULTS: Training enhanced ethical knowing, aesthetic knowing, and empathy. Healthcare professionals also reported increased self-knowledge and skills to use art in group interventions. They gained deeper understanding of loneliness through viewing works of art. CONCLUSION: Viewing art and discussing perceptions of loneliness or suffering was an effective method to help healthcare professionals develop empathy. PMID- 17402382 TI - Complete entrapment of the maxilla. Two different case therapies. AB - Treatment and management of complete entrapment of the maxilla by the mandible can be one of the most difficult orthodontic cases. This affects normal growth and development of the patient. Two entirely different treatment modalities will be demonstrated for correction. PMID- 17402383 TI - The cranial nerve exam. Its importance and how to complete it in 3 minutes. PMID- 17402384 TI - Photographic records for the ortho-TMD practice. PMID- 17402385 TI - What skeletal and dental characteristics do TMD patients have in common? AB - The purpose of this study was to determine if there are any common skeletal or dental characteristics among TMD patients that may be of diagnostic value. Thirty charts of orthodontic patients with pretreatmentTMD symptoms were selected at random.Gender, age, sex, ethnicity, SNA, SNB, ANB,Wits, interincisal angle, missing teeth, prior orthodontic treatment, crossbites, Angle's Class and maxillary and mandibular length were tabulated and analyzed for patterns. The results revealed a clear pattern of excessive mandibular length relative to maxillary length. PMID- 17402386 TI - Effects of oral electrolyte supplementation on endurance horses competing in 80 km rides. AB - REASONS FOR PERFORMING STUDY: There is no evidence that use of oral electrolyte pastes enhances performance in competing endurance horses. OBJECTIVE: To ascertain whether oral administration of a high dose (HD) of sodium chloride (NaCl) and potassium chloride (KCl) to endurance horses would differentially increase water intake, attenuate bodyweight (bwt) loss and improve performance when compared to a low dose (LD). METHODS: A randomised, blinded, crossover study was conducted on 8 horses participating in two 80 km rides (same course, 28 days apart). Thirty minutes before and at 40 km of the first ride 4, horses received orally 02 g NaCl/kg bwt and 0.07 g KCl/kg bwt. The other 4 received 0.07 g NaCl/kg bwt and 0.02 g KCl/kg bwt. Horses received the alternate treatment in the second ride. Data were analysed with 2-way ANOVA for repeated measures (P<0.05). RESULTS: Estimated water intake was significantly greater with HD both at the 40 km mark and as total water intake; however, differences in bwt loss and speed between HD and LD were not found. Treatment significantly affected serum Na+, Cl , HCO3, pH and water intake, but not serum K+ or bwt. Serum Na+ and Cl- were significantly higher at 80 km when horses received HD, but no differences were found in early recovery. Venous HCO3- and pH were significantly lower throughout the ride and in early recovery when horses received HD. CONCLUSIONS AND POTENTIAL RELEVANCE: Other than enhancing water intake, supplementing endurance horses with high doses of NaCI and KCl did not provide any detectable competitive advantage in 80 km rides. Further, the elevated serum electrolyte concentrations induced with HD might not be appropriate for endurance horses. PMID- 17402387 TI - Endurance exercise is associated with increased plasma cardiac troponin I in horses. AB - REASON FOR PERFORMING STUDY: Information is lacking regarding the influence of long distance exercise on the systemic concentration of cardiac troponin I (cTnI) in horses. OBJECTIVES: To determine if the concentration of cTnI in horses competing in 80 and 160 km endurance races increases with exercise duration and if cTnI concentrations can be correlated with performance data. METHODS: Blood samples for the measurement of cTnI and 3 min electrocardiogram recordings were obtained from horses prior to, during and after completion of 80 and 160 km endurance races at 3 ride sites during the 2004 and 2005 American Endurance Ride Conference competition seasons. RESULTS: Full data sets were obtained from 100 of the 118 horses. Endurance exercise was associated with a significant increase in cTnI over baseline in both distance groups. Failure to finish competition (poor performance) was also associated with an increased cTnI concentration over baseline at the time of elimination when data from both distances were combined. Other than one horse that developed paroxysmal atrial fibrillation, no arrhythmias were noted on the 3 minute ECG recordings that were obtained after endurance exercise in either distance group. CONCLUSIONS: Systemic concentrations of cTnI increase in endurance horses competing in both 80 and 160 km distances. Although final cTnI concentrations were significantly increased over their baseline values in horses that failed to finish competition, the degree of increase was not greater than the increase over baseline seen in the horses that successfully completed competition. The clinical significance of increased cTnI in exercising horses could not be ascertained from the results of this study. POTENTIAL RELEVANCE: These data indicate that cardiac stress may occur in horses associated with endurance exercise. Future studies utilising echocardiograpy to assess cardiac function in horses with increased cTnI are warranted. PMID- 17402388 TI - Thyroid hormone responses to endurance exercise. AB - REASONS FOR PERFORMING STUDY: Limited information exists about changes in circulating thyroid hormone concentrations during prolonged endurance exercise in horses. OBJECTIVE: To examine the effects of prolonged exercise on serum iodothyronine concentrations in horses performing endurance exercise of varying distances. METHODS: Serum concentrations of iodothyronines were measured in horses before and after completion of 40, 56, 80 and 160 km endurance rides (Study 1); daily during a 5 day, 424 km endurance ride (Study 2); and before and for 72 h after completion of a treadmill exercise test simulating a 60 km endurance ride (Study 3). RESULTS: In Study 1, 40 and 56 km of endurance exercise had little effect on serum iodothyronine concentrations with the exception of a 10% decrease (P<0.05) in free thyroxine (FT4) concentration after the 56 km ride. In contrast, total thyroxine (T4), total triiodothyronine (T3), FT4 and free triiodothyronine (FT3) concentrations all decreased (P<0.05) after successful completion of 80 and 160 km rides, with decreases ranging from 13-31% and 47-54% for distances of 80 and 160 km, respectively. Further, pre-ride T4 concentration was lower (P<0.05) and FT3 concentration was higher (P<0.05) in horses competing 160 km as compared to horses competing over shorter distances. In Study 2, serum concentrations of T4, T3 and reverse triiodothyronine (rT3) progressively decreased (P<0.05) over the course of the multi-day ride. In Study 3, the greatest decrease (P<0.05) in all iodothyronines was observed at 12 h of recovery, ranging from 25% for FT4 to 53% for FT3, but all thyroid hormone concentrations had returned to the pre-exercise values by 24 h of recovery. CONCLUSION: Endurance exercise results in transient decreases in serum iodothyronine concentrations. POTENTIAL RELEVANCE: These data are important to consider when thyroid gland function is assessed by measurement of serum iodothyronine concentrations in endurance horses. PMID- 17402389 TI - Changes in selected physiological and laboratory measurements in elite horses competing in a 160 km endurance ride. AB - REASON FOR PERFORMING STUDY: Limited information exists about the physiological changes and clinical problems that occur in elite horses competing in high-speed 160 km endurance races. OBJECTIVES: To provide initial data describing changes in physiological and laboratory measurements in horses competing in a high-speed, 160 km endurance race under temperate conditions and to compare data between horses that successfully completed the race and those that failed to finish. METHODS: Body mass (BM) was measured, blood samples were collected, and veterinary examinations performed on horses before, during, and at the finish of a CEI*** 160 km endurance race. RESULTS: Of 36 horses participating in the study, 22 (61%) completed the race. Twelve horses were eliminated for lameness and 2 for persistent heart rate elevation. Mean speed of finishers was 15.2 km/h. Mean +/- s.d. BM loss of finishers at the end of the race (5.7 +/- 2.6%) was not different (P = 0.58) from BM loss of nonfinishers at elimination (6.7 +/- 34%). Similarly, there were no significant differences in heart rate or veterinary assessment of hydration at the race end for finishers as compared to the elimination point for nonfinishers. PCV increased while sodium, chloride and potassium concentrations decreased with exercise but differences between finishers and nonfinishers were not detected. In contrast, both total and ionised calcium concentrations decreased in successful horses but remained unchanged in nonfinishers. CONCLUSIONS: Elite endurance horses are more likely to be eliminated from competition for lameness than metabolic problems; however, it remains unclear whether these conditions are entirely distinct. The magnitude of the decrease in sodium concentration in both finishers and nonfinishers was greater than in previous reports of 160 km rides. POTENTIAL RELEVANCE: These data should be of use for both organisers and participants in elite 160 km endurance races. The tendency toward hyponatraemia as well as the difference in calcium concentrations between finishers and nonfinishers warrant further study. PMID- 17402390 TI - Gene expression profiling in blood cells of endurance horses completing competition or disqualified due to metabolic disorder. AB - REASONS FOR PERFORMING STUDY: Genomics using cDNA microarrays could provide useful information about physiological adaptations and metabolic disorders in endurance horses. OBJECTIVES: In order to show that genes are modulated in leucocytes in relationship with performance and clinical status of the horses, gene expression in leucocytes, haematological and biochemical parameters were compared between successful and disqualified endurance horses. METHODS: Blood samples were collected at rest (TO) and just after a 140-160 km endurance race (T1) in 2 groups of horses: 10 continuing successful (S) and 10 disqualified horses stopped at a vet-gate for metabolic disorders (D). Total RNA was extracted from the blood cells (leucocytes), checked for purity, amplified and hybridised using mouse cDNA microarrays including 15,264 unique genes. Differential gene expressions were studied by hybridisation of each sample T1 vs. a control sample collected at TO (pool of 20 sound horses). RESULTS: Some significant differences were observed in the haematology and biochemistry of the 2 groups (S vs. D). In Group D, rhadomyolysis was confirmed with CK 13,124 u/l and AST 1242 u/l. The list of 726 (including 603 annotated genes) significant genes was filtered according to a high P-value cut-off (P<0.00001). Among them, 130 were upregulated (expression ratio>1.5) and 288 were down-regulated (<1/1.5). Analysis of variance revealed 62 genes differentially expressed (P<0.05) in Groups D and S. The expression levels of 28 and 50 genes were significantly correlated (r>0.75) with CK and AST level in Group D, respectively. The gene ontology classification showed that more genes were up-regulated in S than in the D. More genes were down regulated in the disqualified horses. CONCLUSIONS: Long exercise induced many significant gene modulations in leucocytes. Some genes were expressed in relationship with the clinical phenotype observed in Group D: rhabdomyolysis and haemolysis. POTENTIAL RELEVANCE: Some of these genes could be candidates to explain poor performance or pathologies. Further association studies with a greater number of genes should be conducted. PMID- 17402391 TI - Trot asymmetry in relation to physical performance and metabolism in equine endurance rides. AB - REASON FOR PERFORMING STUDY: Trot is a symmetric gait and asymmetry might appear or increase during endurance rides due to lameness, pain or excessive fatigue. HYPOTHESIS: To assess whether trot asymmetry increases during endurance competitions, whether it is possible to discriminate between horses with different performance and also its possible relationship with metabolism. METHODS: Fifty-eight horses were filmed at trot during the lameness examinations in the vet-gates, before the competition (BCO), and after phases 1 (at 29 km), 2 (at 59 km) and 3 (at the end of the ride, at 80 km) and stride duration (SD) measured in all 4 limbs. A locomotion symmetry index (LSI) was calculated by comparing SD in the diagonal pairs of limbs. In all the cases, the longer diagonal pair SD was compared to the shorter. Horses were classified as symmetric (SyH) when LSI was lower than mean + 2s.d. and asymmetric (ASyH) when LSI was higher than mean + 2s.d. of the data obtained in BCO. Venous samples were withdrawn in the vet-gates and PCV, WBC, creatinine, TPP, uric acid, CK, AST, LDH, Na, K and Cl were measured. Horses were divided into different performance groups: successful (SH), lame (LH) and metabolic (MH), and according to ride velocity, the SH group was also divided into faster finishers (FF) and slower finishers (SF). RESULTS: The percentages of SyH were high (>80%) and very similar between the different performance groups. LSI became more asymmetrical in the vet gates, especially in the LH and MH groups. LSI presented negative correlations with the velocities during the rides and in the vet-gates and CK. The ASyH had higher velocities during the rides and plasma CK and lower velocities during the vet-gates and plasma uric acid concentrations. CONCLUSIONS: Although trot asymmetry increases during endurance events, LSI calculated comparing SD in the two diagonal pair of limbs did not allow the differentiation of horses with different performance. PMID- 17402392 TI - Equine locomotor analysis on vet-gates in endurance events. AB - REASONS FOR PERFORMING THE STUDY: Fatigue during endurance rides might induce changes in gait pattern that could be difficult to distinguish from subtle lameness. OBJECTIVES: To assess changes in gait pattern en route, in fore(FL) and hindlimbs (HL) and establish differences between excellent (EP) and good performers (GP). METHODS: Seventy-five sound and fit horses that completed endurance rides of 80 km were filmed at trot in the vet-gates (VGs) prior to competitions (VGO), and at 29 km (VG1), 59 km (VG2) and 80 km (VG3). Mean gait velocity (V), stride length (SL), duration (SD) and frequency (SF) were measured. Also, stance (StD), swing (SwD), braking (BrD) and propulsion (PrD) durations were determined in the 4 limbs and expressed as % of SD. RESULTS: Horses trotted with a significant lower V at VG3 in comparison to VG0 (mean +/- s.d. 2.3 +/- 0.6 m/sec, 3.3 +/- 0.6 m/sec) and showed a shorter SL (1.5 +/- 0.4 m and 2.2 +/- 0.3 m in VG0 and VG3). Regression analysis confirmed that the reduced SL at VG3 was due to the lower V. Horses in VG3 presented longer FL-StD (46.2 +/- 4.7%, 38.1 +/ 4.6%), BrD (22.0 +/- 2.5%, 16.9 +/- 2.5%) and PrD (24.1 +/- 21%, 21.5 +/- 2.47%) compared to VG0. In VG0, HL-PrD (23.0 +/- 3.0%) and HL-BrD (18.7 +/- 1.8%) were longer than FL-PrD (21.5 +/- 2.47%) and FL-BrD (16.9 +/- 2.5%). EP showed fastest V and longer SL at VG0, and VG3 than GP (P<0.05). CONCLUSIONS: Sound endurance horses undergo changes in gait pattern during competition, mainly due to reduction of V. Some differences exist between FL and HL, and EP show higher V and SL in the VGs. POTENTIAL RELEVANCE: Understanding of the locomotor effect of fatigue will provide an objective basis for the assessment of soundness during endurance events. PMID- 17402393 TI - Stride parameters and hindlimb length in horses fatigued on a treadmill and at an endurance ride. AB - REASON FOR PERFORMING STUDY: The relationship between fatigue and stride and/or muscle stiffness requires further study. OBJECTIVE: To measure stride parameters in horses undergoing fatigue associated with running at submaximal speeds both on a treadmill and in an endurance ride. HYPOTHESIS: Stride frequencies and estimates of hindlimb stiffness would be decreased in fatigued horses. METHODS: Horses were fatigued using 2 paradigms: run to exhaustion at a treadmill (4.5 m/sec, 6% incline) and finishing an 80 km endurance ride. Videos were digitised before and after fatigue and analysed for stride parameters: hind limb length, stride frequency, time of contact, step length, duty factor and stride length. RESULTS: In fatigued horses, stride durations were 5% longer (P = 0.007) resulting in lower stride frequencies (P = 0.016) and longer stride lengths (P = 0.006). The time of contacts (tc) for stance phase were not different (P = 0.108) nor was duty factor (tc/stride period, P = 0.457). Step length (speed x tc) and hindlimb lengths were also not different (P = 0.104, P = 0.8). For endurance horses, stride data for nonfatigued horses were consistent with data extrapolated to 4.5 m/sec from nonfatigued horses on the treadmill. Endurance horses slowed (P = 0.002) during the race from 4.55 to 4.03 m/sec and stride lengths were shorter. Despite a slower speed, other stride parameters were unchanged. Hindlimb length was shorter in fatigued horses. CONCLUSION: Horses fatigued on a treadmill and during the natural course of an endurance ride responded differently, biomechanically. On the treadmill, where speed is constrained, stride frequencies decreased and stride lengths increased. During one endurance ride, stride frequencies were the same, although speeds were substantially reduced. Limb length was shorter in fatigued endurance horses. It remains to be determined if these changes in mechanics are advantageous or disadvantageous in terms of energetics or injury. Further examination of endurance rides is also warranted. PMID- 17402394 TI - A nonsurgical method allowing continuous core temperature monitoring in mares for extended periods, including during endurance exercise. AB - REASONS FOR PERFORMING STUDY: Body temperature can be a useful measure of how well the horse responds to thermal challenges of exercise and environment. Currently there is little information available about how body temperature changes throughout an endurance ride, and no nonsurgical methods to evaluate internal temperature of exercising horses during prolonged periods and during long distances. OBJECTIVE: To develop and test a nonsurgical method, using temperature loggers, of continuously recording internal temperature during exercise and over an extended period. METHODS: Loggers were inserted transcervically into the uterus of mares to measure internal temperature. A total of 9 mares were implanted for periods of 3 days to 6 weeks. The internal temperature was recorded while horses underwent their normal training and competition. Loggers were validated against rectal temperature over a range of internal temperatures while mares were alternatively exercised and rested. RESULTS: The technique proved to be straightforward and accurate. There were no ill effects observed and all mares retained the logger in utero until removal manually, except for one mare, which expelled the logger when she came into oestrus. The intrauterine and deep rectal temperatures were highly correlated and increased during exercise and decreased with aggressive cooling after exercise. CONCLUSIONS: The technique is a useful and accurate nonsurgical method for prolonged monitoring of internal temperature in mares, including those animals in athletic competition. POTENTIAL RELEVANCE: This method allows more intensive studies of core temperature during prolonged exercise in competition and under different environmental conditions. It also enables comparison to be made of different methods of cooling of horses post exercise. PMID- 17402395 TI - Uric acid responses to endurance racing and relationships with performance, plasma biochemistry and metabolic alterations. AB - REASONS FOR PERFORMING STUDY: There is limited understanding of the uric acid response to endurance races. OBJECTIVES: To demonstrate uric acid increments and its relationship to diverse biochemical and performance parameters, in horses subjected to a prolonged effort, with and without presentation of metabolic alterations. METHODS: Blood samples were taken from horses the day before, and 5 10 mins after, successfully finishing a 121 km (Assay 1, n = 24) or 164 km endurance race (Assay 2, n = 17), and from 19 animals eliminated by metabolic disorders during several endurance races (Assay 3). Plasma was obtained and determinations of CK, AST, LDH, AP, uric acid (UA), creatinine (Cr), urea, lactate (La) and plasma proteins (PP) carried out. Sex, age, time in competition, average speed and total recovery time were also recorded. Assays 1 and 2 were arithmetically subdivided into 3 groups each in order to categorise time in competition, average speed and total recovery time. Changes among the groups were evaluated with ANOVA and Fisher's PLSD test. Student's paired t test was used to assess pre- and post exercise differences. A value of P< or =0.05 was considered significantly different. Pearson correlation coefficient was used to assess the relationship between all the variables and UA increases. RESULTS: Average speed of the sampled horses was significantly higher in Assay 1 compared to Assay 2. However, there were no significant differences in plasma biochemistry values between both groups. The fastest horses showed significantly higher UA levels, compared with the slowest (Assays 1 and 2) and medium horses (Assay 1). The animals with alterations in metabolism had significantly higher UA, CK and PP compared with those that adequately concluded the race. There were significant correlations between UA and CK in Assays 1, 2, and 3 and between UA and PP in Assays 1 and 3. CONCLUSIONS: UA rises in horses after a prolonged effort, this increase being higher in animals with metabolic commitment, and in the fastest horses. This increase has a direct correlation with CK. POTENTIAL RELEVANCE: UA could be useful in the assessment of metabolic response during endurance exercise. PMID- 17402396 TI - Does usefulness of potassium supplementation depend on speed? AB - REASONS FOR PERFORMING STUDY: Electrolyte mixtures given to counter sweat loss usually contain abundant potassium. However, increases in plasma [K+] occur with exercise and supplementation may further increase plasma levels, potentially increasing the risk of neuromuscular hyperexcitability and development of adverse clinical sequellae. This proposition requires study. OBJECTIVES: To compare effects of a K-rich electrolyte supplement (EM+K) to a K-free one (EM-K) on plasma [K+], [Ca++] and acid-base status during an endurance incremental exercise test on the treadmill. METHODS: The test consisted of 3 bouts (simulating loops in an endurance race) of 12 km performed at 6, then 7, then 8 m/sec with 25 min rest stops (S1, S2) between loops on 13 endurance trained Arabian horses (7 EM-K, 6 EM+K). Electrolytes were supplied orally 60 mins before exercise (Pre) and at each stop. Blood samples were taken before exercise and during exercise, each S and 120 mins of recovery (R). Blood was analysed for pH, PCO2, packed cell volume (PCV), plasma [Na+], [K+], [Cl-], [Ca++], glucose, and lactate [La-]; plasma [H+] and osmolality (osm) were calculated. The dietary cation anion difference (DCAD) was calculated to be -27 meq/dose EM-K and 109 meq in EM+K, respectively. RESULTS: Plasma [H+] decreased during the 6 and 7 m/sec loops, increased during the 8 m/sec loop, and returned to Pre at S1, S2 and R. Plasma [K+] was higher at 8 m/sec and plasma [Ca++] was overall lower in the EM+K group compared to EM-K. Other findings included higher overall PCV, overall glucose, and [La-] during the 8 m/sec loop (P<0.040) in EM+K compared to EM-K horses. CONCLUSIONS: EM+K supplementation leads to higher plasma [K+] increasing the risk of neuromuscular hyperexcitability during exercise. Acute effects of a lower DCAD in EM-K may have led to higher plasma [Ca++]. Potassium-rich electrolytes may have triggered the release of epinephrine, contributing to higher PCV, glucose release and increased lactate production. POTENTIAL RELEVANCE: Lower plasma [K+] and higher plasma [Ca++] with EM-K supplementation may help reduce the risk of conditions associated with neuromuscular hyperexcitability occurring especially during higher speeds in endurance races. PMID- 17402397 TI - History of horse-whims, teamboats, treadwheels and treadmills. PMID- 17402398 TI - Effect of conditioning horses with short intervals at high speed on biochemical variables in blood. AB - REASONS FOR PERFORMING STUDY: There is limited published work on the effect of training using intensive and short intervals of exercise to condition horses for racing. OBJECTIVES: To examine the effect of conditioning horses 1, 2 or 3 x a week using 2 short fast exercise intervals on blood lactate (LA), plasma ammonia (NH3) and urea (urea) as well as creatine kinase (CK) activity. METHODS: Thoroughbreds (age 4-5 years) were conditioned at near maximal speed (12-14 m/sec) over 100 m, on 2 occasions separated by a 10 min period at walk, on dirt track (conditioning exercise; CE) during a 6 week conditioning period (CP). The CE was undertaken either once (5 horses), twice (5 horses) or 3 times a week (4 horses) during a CP. Before, every 2 weeks during and after the CP, blood was drawn during the CE at 0, 2 and 4 min after each run and additionally 6 min after the 2nd run to measure blood LA and determine the maximal LA post exercise (LAmax). Plasma NH3 was measured in the same samples and the maximal NH3 post exercise (NH3max) determined. Additional blood samples were taken from the horses when stabled before the CE, and 12 and 24 h after to measure urea and CK activity in plasma. RESULTS: There was no differential effect of the number of weekly CE on LAmax, NH3max, LA and NH3 immediately after exercise (LAO and NH3O). Conditioning did not have an effect on LAmax and LA0 after the 1st interval (P>0.05), but LA0 after the 2nd interval was lower after conditioning. NH3max after the 1st and 2nd exercise interval decreased in response to the conditioning, but not NH3 immediately after exercise (P>0.05). Median plasma CK activity 12 h after exercise was higher than before exercise and returned to preexercise levels 24 h post exercise when horses were exercised once and twice/week. In contrast, the median plasma CK activity of horses exercising 3 times/week remained at the pre-exercise level 12 and 24 h post exercise (P>0.05). CONCLUSION: Conditioning horses with 2 intervals of 100 m at near maximal speed had a positive effect on blood LA and plasma NH3. POTENTIAL RELEVANCE: With the type of exercise examined, the fitness of racing horses can be maintained and eventually improved. In further studies the effect of increasing the number of runs in one exercise session should be investigated. PMID- 17402399 TI - The workload of riding-school horses during jumping. AB - REASONS FOR PERFORMING THE STUDY: As there are no reports on the real workload of horses that jump fences, this study was undertaken in riding-school horses. OBJECTIVE: To compare the workload of horses jumping a course of fences with that of horses cantering over the same course at the same average speed without jumping fences. The workload variables included heart rate (HR), packed cell volume (PCV), acid-base balance (venous pH, pCO2, HCO3-) and blood lactate (LA), glucose, total protein and electrolyte concentrations. METHODS: Eight healthy riding-school horses performed test A (a course of approximately 700 m with 12 jumps from 0.8-1.0 m high at an average speed of approximately 350 m/min) and test B (same course at the same speed, but without the rails) in a crossover study with at least 4 h between the 2 tests. Before each test the horses were fitted with a heart rate meter (Polar Electro). Blood samples were taken from the jugular vein at rest prior to the test, after warm-up before starting the course, immediately after the course and after recovery. All samples were analysed immediately. RESULTS: The mean +/- s.d maximal HR (beats/min) during the course (184 +/- 17 and 156 +/- 21, respectively) and the mean HR after recovery (75 +/- 6 and 63 +/- 7, respectively) were significantly higher in test A compared to test B (P = 0.001 and P = 0.007 respectively). The mean LA concentrations after the course and after recovery (mmol/l) were significantly higher in test A (3.6 +/- 2.7 and 1.0 +/- 0.9, respectively) compared to test B (0.9 +/- 0.5 and 0.3 +/ 0.1, respectively), (P = 0.016 and P = 0.048 respectively). The mean PCV (I/l) after the course and after recovery was also significantly different between tests A (0.48 +/- 0.04 and 0.39 +/- 0.03, respectively) and B (0.42 +/- 0.04 and 0.36 +/- 0.03, respectively) (P<0.01). The mean pH and the mean HCO3- (mmol/l) after the course were significantly lower in test A (7.40 +/- 0.04 and 28.9 +/- 1.4, respectively) compared to test B (7.45 +/- 0.03 and 30.4 +/- 2.3, respectively) (P<0.05). CONCLUSIONS: This study indicates that in riding-school horses jumping fences, even at a low level competition, provokes a significant workload compared to cantering the same distance and speed without fences. POTENTIAL RELEVANCE: This study makes it clear that the extra workload of jumping fences should be taken into account in the training programmes of jumping horses. Further research with more experienced horses jumping higher fences will reveal the workload for top-level jumping horses. PMID- 17402400 TI - Body composition in young standardbreds in training: relationships to body condition score, physiological and locomotor variables during exercise. AB - REASONS FOR PERFORMING STUDY: Body composition is an essential factor in athletic performance of human sprinters and long distance runners. However, in horses, many questions remain concerning relationships between body composition and performance in the different equine activities. OBJECTIVES: To determine relationships between body composition, body score, physiological and locomotor variables in a population of young Standardbreds in training. METHODS: Twenty four 2-year-old Standardbreds were studied, body condition on a scale 0-5 and bodyweight recorded, and height at withers measured. Percentage of fat (%F), fat mass (FM) and fat free mass (FFM) were estimated echographically. During a standardised exercise test on the track, velocity, heart rate, respiratory frequency and blood lactate concentrations were measured. V4 and V200 (velocity for a blood lactate concentration of 4 mmol/l and velocity of 200 beats/min) calculated. Basic gait variables were measured at 3 different speeds with an accelerometric device. RESULTS: Body composition variables: %F and FM were significantly related to body condition score and physiological variables. Body score was highly correlated to %F (r = 0.64) and FM (r = 0.71). V4 was negatively correlated to %F (r = -0.59) and FM (r = -0.60), P<0.05. V200 was also negatively related to %F and FM, (r = -0.39 and r = -0.37, respectively, P<0.1). No relationships were found between body composition and gait characteristics. CONCLUSIONS: Body composition was closely related to indirect measurements of aerobic capacity, which is a major factor of athletic performance in middle distance running horses. POTENTIAL RELEVANCE: As in human athletes, trainers should take special note to evaluate optimal bodyweight and body composition of race horses to optimise performance. PMID- 17402401 TI - Optimal active recovery intensity in standardbreds after submaximal work. AB - REASONS FOR PERFORMING STUDY: A retrospective study concerning spontaneous active recovery intensity, i.e. at a freely chosen speed, after a submaximal exercise in trotters showed that the mean intensity demanded by trainers corresponds to 40 50% of maximal heart rate (max HR; unpublished data). However, in human athletes, optimal active recovery intensity was found to be about 60-70% of max HR. Is the spontaneous recovery optimal after a submaximal exercise in trotters? OBJECTIVES: To compare different recovery intensities and define the most efficient one. METHODS: Thirty-seven trotters performed a standardised exercise test on the track. Horses were randomly divided into 4 groups of recovery: passive recovery (n = 10), 10 min walk recovery (n = 10, 100 m/min), 10 min slow trot recovery (n = 9, 250 m/min) and 10 min fast trot recovery (n = 8, 420 m/min). Before, during and 1 h after exercise, speed, heart rate, blood lactate concentration were measured as well as respiratory frequency and rectal temperature. Creatine kinase (CK) was measured 1, 3 and 5 h after exercise. RESULTS: Walk, slow trot and fast trot recovery corresponded respectively to 45-50%, 55-60% and 65-70% of max HR. Heart rate and blood lactate concentration were significantly lower after the 10 sec recovery with increasing intensity of recovery. CONCLUSION: The most efficient intensity of recovery was the 10 min fast trot recovery (65-70% max HR) as this type of recovery allows the optimal blood lactate disappearance. POTENTIAL RELEVANCE: Considering the usual habits of trainers or drivers, recovery intensity after trot races should be increased in intensity to optimise its efficiency. PMID- 17402402 TI - Use of a global positioning and heart rate monitoring system to assess training load in a group of thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: Training is an important variable for determining athletic success. Nonetheless, there has been minimal scientific evaluation of racehorse training programmes. Training of racehorses focuses on running the horses at certain speeds using a combination of a stopwatch and rider's 'feel' for a horse's work intensity. Consequently, actual work intensity for individual horses is not clearly defined. OBJECTIVES: To 1) utilise a combined global positioning system (GPS) and heart rate monitor system to quantify training intensity and physiological responses of a group of racehorses undergoing training and racing; and 2) compare the workload measured by the GPS to that timed and recorded daily by a racehorse trainer. METHODS: Nineteen racehorses age 3 years were followed through a traditional training and racing programme over a 4 month period. Daily GPS and heart rate data together with the trainer's timing and distance data were collected while the horses were trained. Data were analysed using an ANOVA for repeated measures. RESULTS: The combined GPS/heart rate monitoring system detected different heart rate responses in individual horses subjected to the same training workouts. The average speeds detected with the GPS system were in agreement with average speeds timed by the trainer. However, peak speeds reached during training were significantly greater (P<0.05) than those estimated with stopwatch timing. The horses average training speeds increased significantly over the duration of the training period. CONCLUSIONS AND POTENTIAL RELEVANCE: The results from this study show that a GPS/heart rate monitor system provides a reliable measure of daily workload in horses during training. This technology provides a detailed picture of horses' training sessions and has the potential to provide a greater insight into the types of training that may predispose horses to injury. PMID- 17402403 TI - Heart rate responses during acclimation of horses to water treadmill exercise. AB - REASONS FOR PERFORMING STUDY: The use of water treadmill exercise in horses is popular, although little is known about the physiological responses to this form of exercise. No information exists regarding the time taken to acclimate to water treadmill exercise compared to that of high-speed treadmill exercise, for both physiological and biomechanical parameters. OBJECTIVE: To determine heart rate responses during acclimation to water treadmill exercise with and without sedation on first time exposure. METHODS: All horses were exercised on a water treadmill at the walk for 15 min once a day for 4 days. Fourteen horses (mean +/- s.d. age 9 +/- 3.2 years) were assigned randomly to Group A (sedated) and Group B (nonsedated). Group A were given 20-30 microg/kg bwt romifidine within 10-15 min prior to the start of the first acclimating run. Acclimation was determined by the time taken to reach a threshold heart rate value. RESULTS: Group A and B reached threshold heart rate values by the 6th minute of the 4th run (72.8 beats/min) and the 6th min of the 3rd run (78.7 beats/min), respectively. No significant difference was found (P>0.05) between Group A and B in the time taken to reach threshold heart rate values. Acclimation to water treadmill exercise requires a minimum of 2 x 15 min nonsedated acclimating runs. Sedation can be used to prevent horses panicking during the first exposure but thereafter does not affect the time taken to acclimate. POTENTIAL RELEVANCE: Previous experience of water treadmill exercise should be taken into consideration prior to collecting physiological and biomechanical data. Further studies detailing the physiological and biomechanical responses are required prior to making recommendations for the incorporation of this mode of exercise into rehabilitation programmes. PMID- 17402404 TI - Measurements of fitness in thoroughbred racehorses using field studies of heart rate and velocity with a global positioning system. AB - REASONS FOR PERFORMING STUDY: Field based studies of changes in fitness are difficult to conduct in galloping Thoroughbreds. Measurements of heart rate (HR) and real time velocity with a global positioning system (GPS) could provide a method for routine field studies of fitness. OBJECTIVE: To investigate measurements of fitness in the field without using multiple, standardised steps of increasing velocity in the exercise test. METHODS: Twelve healthy 3- to 5-year old Thoroughbreds were used in the study of reliability, and twelve 2-year-old Thoroughbreds in the investigation of the effect of training. Five second averages of velocity and HR were recorded during a typical fast exercise training session, using a GPS and Polar HR monitor. A standardised exercise test protocol was not used. Regression analyses using trot and gallop data were used to calculate velocities at HR of 200 beats/min (V200), and at maximal HR (VHRmax). Data were collected on consecutive 'fast' day training sessions to assess the reliability of measurements. The effect of training was investigated with fitness tests in weeks 2 and 6 of 'fast' training. Absolute and relative differences were calculated to evaluate reliability, and paired t tests were used to detect an effect of training. RESULTS: VHRmax, V200 and HRmax were reliable measurements of fitness, with mean differences of 2% or less. Reliability of VHRmax was not dependent on VHRmax. VHRmax and V200 both increased significantly with training (P<0.01), but there was no effect of training on HRmax. There were no significant changes in the slope or intercept of the regression equations after training. CONCLUSIONS: Velocity and HR measurements during field gallop exercise provided reliable measures of fitness which enabled a measurement of the response to training. POTENTIAL RELEVANCE: This approach offers a simple, noninvasive method for monitoring adaptations to training in the field. PMID- 17402405 TI - Correlation of race earnings with velocity at maximal heart rate during a field exercise test in thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: Running ability of Thoroughbred racehorses is correlated with maximal oxygen uptake, and the velocity at maximal oxygen uptake is highly correlated with the velocity at maximal heart rate (VHRmax). OBJECTIVE: To investigate the relationship between VHRmax and racing performance, expressed as 'peak dollars earned per race start'. METHODS: Heart rate (HR) and velocity were recorded in 25 Thoroughbred racehorses during trotting and subsequent fast gallops in the field at velocities of 15-16 m/sec. Velocity was recorded by a global positioning system (GPS). Maximal HR (HRmax) and maximal velocity (Vmax) were identified, and a linear regression of HR on velocity for trotting and galloping data was constructed to derive VHRmax. Horses followed the training programme designed by one trainer, had at least 6 race starts and were clinically sound at the time of testing. Race earnings were expressed as the peak dollars per start in the horse's race career. Data were normalised using the results for the square root of 'peak dollars earned per race start' and the significance of associations between variables was determined by correlation coefficient and least square analyses. RESULTS: Horses with higher VHRmax earned significantly more dollars per race start (r = 0.41, P<0.05), and horses with VHRmax less than 14.5 m/sec had mean earnings of less than A$2500 per race. There were no correlations between race earnings and either HRmax or Vmax. CONCLUSION: Field studies of the relationship between HR and velocity with a GPS enable identification of horses with limited earnings. POTENTIAL RELEVANCE: This study demonstrates that a field test of fitness of Thoroughbred racehorses that correlates with retrospective racing ability is feasible. The technique has potential application in commercial training environments assisting with decisions concerning racing careers of individual racehorses. PMID- 17402406 TI - The physiological responses to simulated race tests on a track and on a treadmill in standardbred trotters. AB - REASON FOR PERFORMING STUDY: It is unclear to what extent the physiological response to a standardised treadmill exercise test simulating racing conditions resembles the circulatory and metabolic response observed after a simulated race on a track. OBJECTIVES: To compare the physiological responses of a standardised treadmill exercise test used to simulate racing conditions and a simulated race performed on a track on the same Standardbred trotting horses, all in racing condition. METHODS: Six Standardbred trotters in racing condition performed a standardised inclined treadmill exercise test protocol simulating racing conditions (ST) and a simulated race on a field track (FT). Heart and respiratory rates, haemoglobin, packed cell volume (PCV), glucose, pH, total carbon dioxide and potassium in venous blood and plasma lactate and total plasma protein were measured before and immediately after exercise and during recovery. RESULTS: No differences were observed in heart rate, haemoglobin, PCV, total plasma protein, glucose concentrations after exercise and during recovery between the tests. Plasma lactate was higher and total carbon dioxide concentrations and pH were lower in blood at the end of exercise in the FT compared to the ST. Plasma lactate concentrations were still higher 30 min post exercise in the FT compared to the ST. Blood pH returned to resting values at 15 min of recovery for the ST and at 60 min of recovery for the FT. At 60 min of recovery total carbon dioxide concentrations had still not returned to resting values in any of the tests. Respiratory rate at the end of exercise and body temperature at 15 min of recovery was higher after the ST than the FT. Exercise caused an increase in blood potassium concentrations at the end of exercise in both tests, but concentrations were lower after the FT compared to the ST. CONCLUSIONS: The haemodynamic response to the ST test at the end of exercise and during recovery, assessed from heart rate, Hb, and PCV, was similar to the response observed in the FT test. The differences observed in plasma lactate, blood pH and TCO2 concentrations between the ST and FT show that anaerobic metabolism was greater in the FT as this test included a finish at maximal speeds. POTENTIAL RELEVANCE: The treadmill test used in this study to simulate a race resembles the haemodynamic response but not the anaerobic metabolic response observed after a simulated race on a track. PMID- 17402407 TI - Racing speeds of quarter horses, thoroughbreds and Arabians. AB - REASONS FOR PERFORMING STUDY: While Quarter Horses are recognised as the fastest breed of horse, direct comparisons to race times with other breeds can be misleading. Quarter Horse races begin when the starting gates open. Thoroughbred and Arabian races begin a short distance from the gates after horses have started running. This study compared speeds of these breeds as they accelerate from the starting gates and during the middle and end of races. OBJECTIVES: To compare racing speeds of the 3 breeds, and to compare speeds during various segments of the races. METHODS: Video tapes of races were obtained from a local track. The various race segments were viewed and the winning horse timed by 5 individuals. Fastest and slowest times were removed and the 3 remaining times averaged. RESULTS: Quarter Horses averaged faster speeds than Thoroughbreds even when Thoroughbreds were raced at a distance (402 m) similar to Quarter Horse races. Both breeds were substantially faster than Arabians. Quarter Horses racing 336 m or less gained speed in each segment of the race while Arabians and Thoroughbreds racing 1006 m ran fastest during the middle of the race and had decreased their speed in the final segment of the race. CONCLUSIONS: Despite similar race times reported for 402 m, Quarter Horses averaged faster speeds than Thoroughbreds when timed from a standing start. In short races, both breeds accelerate throughout the race. Arabians, despite being known for endurance, had slowed by the end of the race. POTENTIAL RELEVANCE: This study demonstrates that Quarter Horses achieve faster racing speeds than do other breeds. It also reveals a potential flaw in race-riding strategy as a more consistent pace throughout the Arabian and longer Thoroughbred races may be more efficient and result in a faster overall race time. PMID- 17402408 TI - A comparison of cross-country recovery rates at CCI 2* with and without steeplechase competitions. AB - REASONS FOR PERFORMING STUDY: Short format 3-day events were introduced in 2004. Anecdotal reports suggested that horses were more tired on completion of the cross-country phase of short format events when compared with horses completing the cross-country phase of long format competitions, despite the absence of Phases A, B and C. OBJECTIVES: To compare the physiological parameters and haematological parameters of horses that had completed the cross-country phase of a short format (SF) and a long format (LF) CCI 2* competition. METHODS: During a CCI 2* competition 69 competitors took part in the short format and 74 in the long format competition. Long format competitors completed Phases A, B, C and D and short format competitors completed Phase D only. Phase D (the cross-country course) was identical for both competitions. Two-way ANOVA for repeated measures and post hoc tests were used to compare temperature, pulse and respiration rates of horses competing in both types of competition. T tests were used to compare mean lactate and electrolyte concentrations, while U-Mann Whitney tests were used to compare CK and AST levels measured in horses competing in the short and long formats of the event. RESULTS: Training schedules, age and previous competition experience were not significantly different between horses competing in the SF and LF competitions. On completion of Phase D, SF horses had significantly higher PCV and significantly lower ionised calcium concentrations when compared with LF horses. LF horses had significantly higher heart rates than SF horses 10 min prior to starting Phase D and immediately after completing Phase D; however, no other significant differences were found between the 2 groups of horses. CONCLUSIONS: Only weak evidence was found to support the hypothesis that the workload for the horse in a SF CCI 2* competition is significantly different when compared to the LF CCI 2* competition. POTENTIAL RELEVANCE: There is no beneficial or detrimental effect on horses that complete short format CCI 2* competitions as compared to those that complete long format CCI 2* competitions but further research is required into the physiological response of horses at CCI 3* and CCI 4* short format competitions. PMID- 17402409 TI - Utilisation of the time constant calculated from heart rate recovery after exercise for evaluation of autonomic activity in horses. AB - REASONS FOR PERFORMING STUDY: Heart rate (HR) recovery immediately after exercise is controlled by autonomic functions and the time constant (T) calculated from HR recovery is thought to be an index of parasympathetic activity in man. OBJECTIVES: To investigate whether it is possible to evaluate autonomic function using the time constant in horses. METHODS: Five Thoroughbred horses were subjected to a standard exercise test. Following pre-medication with saline, atropine and/or propranolol, the horses ran for 2.5 min at a speed of 8 m/sec at a 10% incline and T was calculated from HR after the exercise. Secondly, 7 Thoroughbred horses were then trained for 11 weeks and T and maximal oxygen uptake (VO2max) measured at intervals of 1 or 2 weeks. In 6 horses, T with atropine pre-medication was also measured before and after the whole training period. Furthermore, the HR variability at rest was evaluated by power spectral analysis at intervals of 3 or 4 weeks. RESULTS: Time constant was increased by atropine and/or propranolol pre-medication, decreased with the progress of training and inversely correlated with VO2max during training (r = 0.43, P<0.005). Parasympathetic blockade significantly decreased T only after and not before, the training; however, T was lower in post training than in pretraining, irrespective of parasympathetic blockade. On the other hand, parasympathetic activity at rest was attenuated and sympathetic activity became predominant following the training. CONCLUSION: Heart rate recovery is affected by sympathetic withdrawal and parasympathetic reactivation in horses and suggests that physical training hastened HR recovery by improving the parasympathetic function after exercise with aerobic capacity. However, the effects of other factors need to be considered because the training effect appeared on T even under parasympathetic blockade. The parasympathetic activity at rest is in contrast to that after exercise, suggesting that T does not reflect parasympathetic activity at rest. POTENTIAL RELEVANCE: If demonstrated how HR recovery is controlled after exercise, its analysis will be important in the evaluation of physical fitness in horses. PMID- 17402410 TI - Retrospective study of predictive variables for maximal heart rate (HRmax) in horses undergoing strenuous treadmill exercise. AB - REASONS FOR PERFORMING STUDY: Heart rate is one of the most commonly measured variables in equine exercise physiology and relative exercise intensity commonly expressed as % of maximal heart rate. A number of influences affect maximal heart rate (HRmax), including age of the horse but other factors have not been described. OBJECTIVES: To determine if fitness, health status, gender, breed, athletic use, body mass, in addition to age, are predictive of HRmax in the horse. METHODS: Maximal heart rate data from 328 horses which underwent treadmill exercise tests at 5 different laboratories were obtained retrospectively. Univariable linear regression analyses were performed on individual variables. Multiple linear regression analysis using a backward elimination modelling procedure was then used to relate the observed HRmax values simultaneously with different predictive variables. Variables were retained in the final regression model if they or any of their categories were significantly predictive of HRmax at P<0.05 and if there was a significant collective contribution to the model from inclusion of each variable, also at P<0.05. RESULTS: Age, fitness status, laboratory, gender and breed/use (combined category) were all statistically significantly predictive of HRmax. Together these variables accounted for 41% of the variance in HRmax. Age alone accounted for only approximately 13% of the variation between horses in HRmax. Neither body mass nor health status were significantly predictive. CONCLUSIONS: HRmax in the horse declines with age but is also influenced by other factors. As the factors investigated accounted for only 41% of the variation between horses, other unidentified variables with a strong influence on HRmax remain to be identified. POTENTIAL RELEVANCE: Factors such as fitness, age, gender, breed and use need to be considered when interpreting estimates or measurements of HRmax. PMID- 17402411 TI - An echocardiographic and auscultation study of right heart responses to training in young national hunt thoroughbred horses. AB - REASONS FOR PERFORMING STUDY: There are few data available to determine the effect of training on cardiac valve function. OBJECTIVES: To investigate the effect of commercial race training on right ventricular (RV) and tricuspid valve function in an untrained group of National Hunt Thoroughbreds (TB). MATERIAL AND METHODS: Cardiac auscultation, guided M-mode echocardiography of the RV, and colour flow Doppler (CFD) tricuspid valve and right atrium were performed in 90 TB horses (age 2-7 years) 1998-2003. Forty horses were examined at least once and 48 horses were examined on at least 2 occasions. Examinations were then classified as: i) before commencement of race training, ii) after cantering exercise had been sustained for a period of 8-12 weeks and iii) at full race fitness. Tricuspid valve regurgitation (TR) murmurs were graded on a 1-6 scale and CFD echocardiography TR signals were graded on a 1-9 scale. Right ventricular internal diameter (RVID) in diastole and systole (RVIDd and RVIDs) was measured by guided M-mode. Associations between continuous RVID and TR measures and explanatory covariates of weight, age, heart rate, yard and stage of training were examined using general linear mixed models with horse-level random effects. RESULTS: On average, RVIDd and RVIDs increased by 0.08 and 0.1 cm, respectively, per year increase in age (P = 0.1 and 0.02) and by 0.3 and 0.4 cm, respectively between pretraining and race fitness (P = 0.07 and 0.005). Tricuspid regurgitation score by colour flow Doppler increased by 0.6/year with age (P<0.0001) and by 1.8 between pretraining and race fitness (P<0.0001). No significant associations were found between any outcomes and weight, heart rate and training yard. Due to the high level of colinearity between age and training, multivariable models including both terms were not interpretable. CONCLUSIONS AND CLINICAL RELEVANCE: Athletic training of horses exerts independent effects on both severity and prevalence of tricuspid valve incompetence. This effect should therefore be taken into account when examinations are performed. Dimensions of RV increase with age and training in TB horses in a manner that appears to be similar to that of the LV. PMID- 17402412 TI - Exercise and pharmacological stress echocardiography in healthy horses. AB - REASONS FOR PERFORMING STUDY: Stress echocardiography could be a useful diagnostic test in horses suspected to suffer from exercise-induced myocardial dysfunction as a cause of exercise intolerance. OBJECTIVE: To evaluate the effect of treadmill exercise and pharmacological stress test on left ventricular echocardiographic parameters. METHODS: Echocardiography was performed in 2 groups of 5 healthy horses, either immediately after a near-maximal treadmill exercise (Group EXE) or during a pharmacological challenge (Group DOB) consisting of 35 microg/kg atropine, followed by incremental dobutamine infusion rates of 2-6 microg/kg bwt/min for 5 min duration each, which led to a cumulative dobutamine dose of 100 microg/kg. Left ventricular M-mode parameters were compared at rest and at heart rates of 80, 100, 110, 120 130, and 140 beats/min, within each group. RESULTS: In 2 horses of Group EXE, echocardiographic measurements were impossible at 140 and 130 beats/min, as their heart rates dropped too fast in the immediate post exercise period. In the remaining 3 horses image quality was not always satisfactory at heart rates of 130 and 140 beats/min. Systolic left ventricular parameters and fractional shortening measured at 130 and 140 beats/min were significantly different from values obtained at lower heart rates. Horses in Group DOB reached expected heart rates of 80 and 100 beats/min, after the administration of atropine and during a dobutamine infusion rate of 2 microg/kg bwt/min, respectively. Heart rates targets of 110, 120, 130, and 140 beats/min were reached at mean (+/- s.d.) dobutamine infusion rates of 2.8 +/- 0.4, 3.2 +/- 0.4, 4.0 +/- 0.7, 5 +/- 0.7 microg/kg bwt/min, respectively. Systolic left ventricular parameters and fractional shortening at heart rates of 110, 120, 130, and 140 beats/min, were significantly different from values obtained at lower heart rates. CONCLUSION: The pharmacological stress test induced changes in ventricular dimensions at heart rates of 80 to 140 beats/min. Using this test, high quality images can easily be obtained at heart rates of 140 beats/min. Conversely, in post exercise echocardiography, obtaining good quality images at heart rates of 130 and 140 beats/min is difficult, which limits use of the technique in routine clinical settings. POTENTIAL RELEVANCE: Further studies should demonstrate the potential of pharmacological stress test as a diagnostic tool in horses suffering from exercise-induced myocardial dysfunction. PMID- 17402413 TI - Cardiac arrhythmias during and after treadmill exercise in poorly performing thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: The prevalence and severity of cardiac arrhythmias during exercise in athletic horses presented for poor performance is not well described. OBJECTIVES: To describe prevalence and severity of ventricular and supraventricular arrhythmias immediately before, during and immediately after standardised incremental treadmill exercise tests (IET) to fatigue in Thoroughbred horses during investigation of poor performance. METHODS: The electrocardiograms (ECG) of 88 Thoroughbred racehorses, judged to be free of significant heart disease or arrhythmia at rest, were used. A modified base-apex ECG was recorded throughout an IET to fatigue. Recordings were analysed independently by 2 observers. Twenty-eight horses were diagnosed with dorsal displacement of the soft palate, 25 had varying degrees of soft palate instability and aryepiglottic fold collapse, 8 had other respiratory problems and, in 27 cases, no definitive diagnosis was reached. RESULTS: Fifty-five horses had at least one ventricular (VPD) or supraventricular (SVPD) depolarisation, 23 had only VPDs, 17 had only SVPDs and 15 had both in at least one exercise period. Premature depolarisations were seen predominantly during the first min of recovery from IET. The range of premature beats after exercise was 1-30 VPDs, and 1-9 SVPDs. No significant associations were observed between age, sex, race type, diagnosis, peak heart rate or run time to fatigue during IET and occurrence of either > or =1 premature beat or of more severe arrhythmias (multiple singles [>5] or pairs or paroxysms of premature depolarisations during peak exercise or immediately after exercise). However, a larger sample size would be required to have greater confidence in these associations. CONCLUSIONS: Isolated VPDs and SVPDs are frequently detected in poor performing racehorses during IET but their clinical relevance remains to be determined. POTENTIAL RELEVANCE: The guidelines for interpretation and clinical relevance of premature depolarisations observed during and immediately after treadmill exercise tests in poor performing Thoroughbred racehorses deserves further evaluation. PMID- 17402414 TI - Measurements of right ventricular internal dimensions and their relationships to severity of tricuspid valve regurgitation in national hunt thoroughbreds. AB - REASONS FOR PERFORMING STUDY: The effect of tricuspid valve regurgitation (TR) in right ventricular (RV) chamber size has not been reported. METHODS: An echocardiographic and auscultation study was conducted on 394 race-fit National Hunt Thoroughbreds (TBs) in 4 training yards. Auscultation was performed on each horse and audible murmurs of TR graded using a 1-6 scale. Echocardiography was also performed and standard, 2-dimensional (2D), short (SA) and long axis (LA) images of the RV obtained from a right parasternal location. An m-mode image was recorded from an SA view of the RV just below the level of the tricuspid valve. Colour flow Doppler (CFD) echocardiography was used to interrogate the tricuspid valve and right atrium. If retrograde flow was visible, representative recordings were also acquired. Severity of TR by CFD was graded using a subjective 1-9 scale. Measurements of RV chamber size in systole and diastole were made from archived 2D, LA, SA and m-mode images by a single observer unaware of the auscultation and CFD findings. To determine day-to-day repeatability of RV measurements from each imaging plane and of TR by CFD, 5 horses were examined on 5 consecutive days using an identical echocardiographic protocol. Images from 30 horses in the original dataset were also measured on a second occasion to determine the intra-observer repeatability. RESULTS: There were significant positive correlations between the severity of TR by CFD and RV internal diameter from each image plane. In contrast, there were no significant associations between grade of TR by auscultation and any RV dimension. Data also showed that only RV measurements derived from M-mode images have acceptable intraobserver repeatability and similarly the M-mode measurements had the best overall day-to day repeatability. CONCLUSIONS AND CLINICAL RELEVANCE: Data showed that TR assessed by CFD was positively associated with the diastolic and systolic dimensions of the RV, but failed to demonstrate a positive association between the presence of a murmur of TR and RV chamber size. M-mode and 2D echocardiography provided moderately reliable and repeatable methods for obtaining measurements of RV internal dimensions. M-mode images provide the most reliable and repeatable measurements, particularly for less experienced operators. PMID- 17402415 TI - Effect of light exercise on valvular regurgitation in standardbred trotters. AB - REASONS FOR PERFORMING STUDY: Although valvular regurgitation is diagnosed frequently by colour Doppler echocardiography in racehorses at rest, there is currently no information about the effect of exercise on valvular regurgitation. OBJECTIVES: To investigate the effect of light exercise on valvular regurgitation in a population of trotters. METHODS: Standardbred trotters (n = 73) were examined by resting auscultation and echocardiography, of which 50 fullfilled the criteria of the study. Of these, 4 had reduced performance according to the trainers and were not included in the statistical analyses. Horses that showed valvular regurgitations diagnosed by colour Doppler echocardiography at rest, were light exercised and the regurgitant valves were then re-examined by colour Doppler echocardiography. RESULTS: Of the 46 horses, 41 (89%) had one or more valvular regurgitations at rest with tricuspid regurgitation present in 83% of horses, mitral regurgitation 46%, pulmonary regurgitation 37% and aortic regurgitation 57%. After exercise, one or more valvular regurgitation persisted for 23 of the 41 horses (56%). The regurgitation decreased significantly for mitral, pulmonary and aortic valves; the mitral regurgitation persisted for 21% of the horses, pulmonary regurgitation persisted for 6% and aortic regurgitation persisted after exercise for 23 % with regurgitation diagnosed at rest. Half of those with tricuspid regurgitation at rest still had tricuspid regurgitation after exercise. CONCLUSIONS: The results of this study showed that the majority of small valvular regurgitations disappear after light exercise. However, the severity of tricuspid regurgitation increased in 4 horses. Furthermore, these horses were not performing satisfactorily according to the trainers. POTENTIAL RELEVANCE: Studies in man have shown that severity after exercise suggests pathological regurgitation. The present study indicates that echocardiographic examination of valvular regurgitation after exercise might be a relevant examination in horses. However, more studies are needed to evaluate the effect of strenuous exercise on valvular regurgitation in horses. PMID- 17402416 TI - Right ventricular pressure dynamics and stress echocardiography in pharmacological and exercise stress testing. AB - REASONS FOR PERFORMING STUDY: There is interest in using pharmacological stress testing (PST) as a substitute for exercise stress testing (EST) to evaluate cardiac function in horses. OBJECTIVES: To compare the effect of PST and EST on right ventricular pressure dynamics and stress echocardiography. METHODS: Five horses completed a PST and EST in a randomised crossover design. High fidelity pressure transducers were placed in the right ventricle. Continuous pressure signals were digitally collected and stored, and dP/dtmax, dP/dtmin and tau calculated from these measurements. ECGs were recorded continuously for 20 h. Echocardiography was performed prior to EST and PST, during and after PST, and immediately post EST. Plasma cardiac troponin I concentrations were measured pre- and 3-4 h post stress testing. For PST, 5 microg/kg bwt glycopyrrolate i.v. followed after 10 min by 5 microg/kg bwt/min dobutamine infusion over 10 min was given. EST consisted of a 2 min gallop at 110% speed required to elicit VO2max. RESULTS: Both EST and PST resulted in a significant increase in right-ventricular dP/dtmax and dP/dtmin over baseline (P<0.05) and a significant decrease in tau compared with baseline (P<0.05). EST dP/dtmax and dP/dtmin were significantly greater than PST dP/dtmax and dP/dtmin (P<0.05) and EST tau was significantly less than PST tau (P<0.05). Two minutes post EST and 5 min post PST dP/dtmax were not significantly different, but were significantly less than end-EST and during PST. Tau was also not significantly different between post EST and post PST, but was significantly decreased end-EST compared with during PST. FS were not significantly different between PST and post EST, but during PST and post EST all FS were significantly higher than baseline. Cardiac troponin I concentrations were significantly elevated post PST and were greater than post EST. The clinical relevance of this is unknown. CONCLUSIONS: PST had a similar, although less marked effect on the cardiac parameters related to right-ventricular pressure dynamics and a similar effect on echocardiography as exercise stress testing. POTENTIAL RELEVANCE: PST deserves further evaluation in normal horses and those with cardiac disease, and may be complementary to EST to better identify exercise induced cardiac dysfunction. PMID- 17402417 TI - Associations between exercise-induced pulmonary haemorrhage, right ventricular dimensions and atrioventricular valve regurgitation in conditioned national hunt racehorses. AB - REASONS FOR PERFORMING STUDY: Exercise-induced pulmonary haemorrhage (EIPH) and tricuspid (TR) and mitral valve regurgitation (MR) are conditions with high prevalence in conditioned Thoroughbreds. Stress failure of pulmonary capillaries, leading to EIPH, is most likely when pulmonary vascular pressure is increased, and mitral regurgitation can result in pulmonary venous hypertension. HYPOTHESIS: There might be an association between MR and EIPH and the right ventricle (RV) of horses known to suffer repeated episodes of EIPH, as their hearts would be subject to higher pulmonary vascular pressures and cardiac output during training and hence increased RV afterload and preload. METHODS: An echocardiographic and auscultation study was conducted in 121 race-fit National Hunt Thoroughbreds. Cardiac auscultation and echocardiography were performed. A guided M-mode image of the RV just below the tricuspid valve was obtained from a right parasternal location and colour flow Doppler (CFD) used to interrogate the tricuspid valve and right atrium. The mitral valve was similarly examined from the left hemithorax. Severity of TR and MR by CFD was graded. Binary data on EIPH, based on whether the horse was perceived to have a clinically significant problem with EIPH, were determined retrospectively for each horse by the horses' primary care veterinary surgeon from medical and other records. Data were analysed using a standard logistic regression analysis approach. RESULTS: EIPH was significantly and positively associated with the systolic and diastolic dimensions of the RV (P = 0.017 and 0.011 respectively) and this association was not sensitive to the effects of age or weight. There were no significant associations between EIPH and TR or MR by auscultation or CFD (TR: auscultation P = 0.1; CFD P = 0.2 and MR: auscultation P = 0.07; CFD P = 0.37). CONCLUSIONS AND CLINICAL RELEVANCE: This study was limited by the method used to classify EIPH, but there was no association between EIPH and horse age, weight, TR or MR in this population of Thoroughbreds. Nevertheless, RV internal dimension was greater in horses obviously affected by EIPH, suggesting that factors resulting in EIPH may directly or indirectly affect RV remodelling in athletic horses. PMID- 17402418 TI - Effects of a specific endothelin-1A antagonist on exercise-induced pulmonary haemorrhage (EIPH) in thoroughbred horses. AB - REASONS FOR PERFORMING STUDY: During high intensity exercise, the very high pulmonary artery pressure (Ppa) experienced by Thoroughbred horses is considered a major factor in the aetiology of exercise-induced pulmonary haemorrhage (EIPH). Recently, endothelin-1 (ET-1), a potent vasoconstrictive hormone, has been found to increase Ppa in horses at rest via binding to its ET-1A receptor subtype. In addition, plasma concentrations of ET-1 are increased in horses during and after high intensity exercise. HYPOTHESIS: If ET-1 increases Ppa during exercise in the horse, administration of a specific ET-1A antagonist would decrease Ppa and therefore EIPH. METHODS: Saline (CON) or an ET-1A receptor antagonist, TBC3214 (3 mg/kg bwt i.v.; ANTAG) was administered to horses 1 h prior to maximal incremental exercise on a high-speed treadmill. Gas exchange measurements were made breath-by-breath and blood samples collected during each 1 min stage to determine blood gases, acid-base status and cardiac output. EIPH was determined via bronchoalveolar lavage (BAL) approximately 30 min after exercise. RESULTS: The time to fatigue, gas exchange and cardiovascular responses were not different between groups (P>0.05). Resting and peak Ppa did not differ significantly between treatments. Most importantly, ANTAG did not decrease EIPH. CONCLUSIONS: These results do not support a deterministic role for ET-1 in the increased Ppa and therefore EIPH, during maximal exercise in the equine athlete. POTENTIAL RELEVANCE: Treatment with an ET-1A receptor antagonist does not appear to be a viable therapeutic intervention in the prevention of EIPH. PMID- 17402419 TI - Effect of repeated exercise and recovery on heart rate variability in elite trotting horses during high intensity interval training. AB - REASONS FOR PERFORMING STUDY: Interval training is a commonly used training method for trotting horses. In addition, trainers are provided with efficient and inexpensive heart rate monitor devices for the management of training. HYPOTHESIS: Since the high frequency (HF) frequency peak (fHF) of heart rate variability (HRV) corresponds to the breathing frequency in combination with stride frequency during trotting, it is hypothesised that modifications of breathing and stride frequencies induced by repeated exercise could be detected from fHF. METHODS: RR interval time series of 7 trotting horses were recorded during an interval training session. Interval training was made up of 5 successive 800 m high-velocity trotting runs (H1, H2...H5) separated by 1 min recovery bouts at low speed (R1, R2...R5). Fast Fourier transform (FFT) and Poincare plot analysis techniques were applied to RR series. RESULTS: Repeated exercise had significant effects on HRV components during interval training. Despite constant trotting velocities during high-speed and recovery, repetition induced a decrease in mean RR interval (H1: 295 +/- 19 vs. H5: 283 +/- 15 msec, P<0.05) and in the root mean square of successive differences in RR series (RMSSD; H1: 6.31 +/- 1.28 vs. H5: 5.31 +/- 1.31 msec, P<0.05). Furthermore, high speed and recovery repetitions induced an increase in fHF (H1: 1.37 +/- 0.35 vs. H5: 1.62 +/- 0.40 Hz and R1: 0.22 +/- 0.02 vs. R4: 0.64 +/- 0.38 Hz, P<0.05). Hence, recovery induced a decrease in the s.d. of the successive RR series (SDRR; R3: 10.5 +/- 3.96 vs. R5: 6.17 +/- 2.65 msecs, P>0.05) and in the long term index of Poincare plot (SD2; R1: 43.29 +/- 28.90 vs. R5: 18.19 +/- 9.35 msecs, P<0.05). CONCLUSIONS: The observed increase in fHF during the interval training could be induced by alterations of the coupling between breathing and stride frequency linked to the emergence of fatigue. The decrease in SD2 and SDRR during successive recovery bouts could be linked with a deterioration of the recovery pattern. POTENTIAL RELEVANCE: HRV can provide breathing frequency data of Standardbreds during training without any respiratory device. Furthermore, HRV could provide useful makers of the emergence of fatigue states during training. PMID- 17402420 TI - Effect of detraining on cardiorespiratory variables in young thoroughbred horses. AB - REASONS FOR PERFORMING STUDY: Thoroughbred racehorses often experience interruptions to their training. Identifying the effects of these changes and how they alter athletic performance might provide an insight on to how to prevent these changes from occurring. HYPOTHESIS: Training and detraining young Thoroughbreds alters their aerobic capacities with correlated changes in circulatory capacities; if horses remained spontaneously active in a pasture during their detraining period, their decreases in aerobic capacity during detraining would be reduced. METHODS: We trained 6 Thoroughbred yearlings for 6 months using a conventional yearling race training programme. They were then detrained for 10 weeks with free range on pasture for 8 h/day and stall rest at night. Treadmill measurements of O2 transport variables were made before training (PRE), after training (TR) and after detraining (DT). A step-test protocol identified each horse's aerobic capacity (VO2max) and speed to attain it, and a steady-state run at VO2max was used to quantify 02 transport variables at each time period. RESULTS: The mass-specific and whole-body VO2max, cardiac output (Q) and stroke volume (Vs) increased from PRE to TR. All mass-specific values decreased significantly from TR to DT; however, because body mass increased by 8.3% from TR to DT, none of the variables changed significantly from TR to DT on a whole-body basis. CONCLUSIONS: Changes in aerobic capacity are highly correlated with changes in Vs and circulatory capacity during training and detraining. Exercise activity of trained young horses free at pasture for 8 h/day is sufficient to maintain VO2max, (Q and Vs during 10 weeks of DT. POTENTIAL RELEVANCE: Aerobic and cardiovascular fitness may be maintained in young Thoroughbred horses during at least 10 weeks of detraining by maintaining modest spontaneous exercise activity. PMID- 17402421 TI - Heart rate variability in trotters during different training periods. AB - REASONS FOR PERFORMING STUDY: Endurance training induces changes in autonomic nervous system functions. High intensity training includes the risk of overtraining, in man and horse. Heart rate variability (HRV) is a noninvasive measurement of the autonomic regulation of the heart rate, which is quick and easy to measure with modern telemetric technology. HYPOTHESIS: Since HRV is affected by changes in the autonomic nervous system, it might be an early stage indicator of poor recovery from a previous bout of exercise or overreaching or overtraining in horses in general. METHODS: The aim of the study was to monitor recovery and the possible overtraining status in horses by measuring HRV. The measurements reflected the responses of the previous day activities during different training periods including basic training, precompetition and competition during a one-year follow-up. RESULTS: HRV was at the highest during precompetition period (P<0.05) and it decreased significantly during competition period (P<0.05), indicating an increased stress load in the competition period. Walking increased HRV significantly compared to complete rest or jogging as previous day activities during basic training and precompetition periods (P<0.05). This finding suggests that horses are more relaxed during moderate exercise than standing still or anaerobic exercise. CONCLUSIONS: HRV can be used to monitor the cardiovascular responses to training in horses but confirmatory measures may also be required in addition to HRV to exclude other possible causes of underperformance. PMID- 17402422 TI - The effect of long-term exercise on glucose metabolism and peripheral insulin sensitivity in standardbred horses. AB - REASONS FOR PERFORMING STUDY: To study the possible long-term effect of improved glucose tolerance in horses after long-term training, as the impact of exercise training on glucose metabolism is still unclear in the equine species. It is not known whether there is a direct long-term effect of training or if the measurable effect on glucose metabolism is the residual effect of the last exercise session. OBJECTIVES: To determine the chronic effect on glucose metabolism and peripheral insulin sensitivity of long-term training in horses by use of the euglycaemic hyperinsulinaemic clamp technique. METHODS: Eleven Standardbred horses were acclimatised to running on the high-speed treadmill for 4 weeks (Phase 1) followed by training for 18 weeks with an alternating endurance (approximately 60% HRmax) high intensity training programme (approximately 80% HRmax) (Phase 2). Training frequency was 4 days/week. At the end of Phase 1, a euglycaemic hyperinsulinaemic clamp was performed 72 h after the last bout of exercise in all horses. At the end of Phase 2, the horses were clamped 24 h or 72 h after the last bout of exercise. RESULTS: Glucose metabolism rate did not change significantly after 18 weeks of training, measured 72 h after the last exercise bout (0.018 +/- 0.009 and 0.022 +/- 0.006 mmol/kg bwt/min, respectively). Peripheral insulin sensitivity also did not change significantly following training (7.6 +/- 5.7 x 10(-6) and 8.0 +/- 3.1 x 10(-6), respectively). The same measurements 24 h after the last bout of exercise showed no significant differences. CONCLUSIONS: Results indicated that long-term training in Standardbreds neither changed glucose metabolism or insulin sensitivity 72 h after the last bout of exercise. POTENTIAL RELEVANCE: The fact that the beneficial effect of increased insulin sensitivity after acute exercise diminishes quickly in horses and no long-term effects on insulin sensitivity after chronic exercise have as yet been found in horses, implies that exercise should be performed on a regular basis in horses to retain the beneficial effect of improved insulin sensitivity. PMID- 17402423 TI - Effects of short-term training on insulin sensitivity and skeletal muscle glucose metabolism in standardbred horses. AB - REASONS FOR PERFORMING STUDY: Increased insulin sensitivity occurs after a period of exercise training, but the mechanisms underlying this training-associated increase in insulin action have not been investigated. OBJECTIVE: To examine the effects of short-term endurance training (7 consecutive days) and a subsequent period of inactivity (5 days) on whole body insulin sensitivity and GLUT-4 protein and the activities of glycogen synthase (GS) and hexokinase (HK) in skeletal muscle. It was hypothesised that training would increase insulin sensitivity in association with increased GLUT-4 protein and activities of GS and HK, but that these changes would be transient, returning to baseline after 5 days of inactivity. METHODS: Seven mature Standardbred horses completed training consisting of 7 consecutive days of 45 min of treadmill exercise at a speed that elicited 55% of pretraining maximal aerobic capacity (VO2peak). Insulin sensitivity was determined by rate of glucose disposal (M) during the last 60 min of a 120 min euglycaemic-hyperinsulinaemic clamp (EHC) performed before (-2 days) and at 1 and 6 days following training. VO2peak was measured before (UT) and after (TR) training and the period of inactivity (IA). RESULTS: Training resulted in a 9% increase in mean VO2peak (P<0.05) that was maintained following inactivity (IA). Mean M values were more than 2-fold higher (P<0.05) in TR than in UT. Mean M was also higher (P<0.05) in IA when compared to UT. GLUT-4 protien abundancewas more than 10-fold higher in TR and IA (P<0.001) than in UT. Pre-EHC GS activity and GS fractional velocity were increased (P<0.05) in TR when compared to UT and IA. Pre-EHC HK activity was increased (P<0.05) in IA when compared to UT and TR. Muscle glycogen was 66% lower (P<0.05) in TR than in UT and IA. CONCLUSIONS: Short-term training resulted in increases in whole body insulin sensitivity, and GLUT-4 protein content and glycogen synthase activity in skeletal muscle. The enhancements in insulin sensitivity, GLUT-4 protein and glycogen synthase activity were still evident after 5 days of inactivity. POTENTIAL RELEVANCE: Insulin resistance in equids has been associated with obesity and predisposition to laminitis. Regular physical activity may mitigate risk of these conditions via enhancement of insulin sensitivity and/or control of bodyweight. PMID- 17402424 TI - Low dose exogenous erythropoietin elicits an ergogenic effect in standardbred horses. AB - REASONS FOR PERFORMING STUDY: Recombinant human erythropoietin (rhuEPO) causes an increase in red blood cell production and aerobic capacity in other species; however, data are lacking on effects in the horse. HYPOTHESIS: This study tested the hypothesis that rhuEPO administration would alter red cell volume (RCV), aerobic capacity (VO2max) and indices of anaerobic power. METHODS: Eight healthy, unfit mares accustomed to the laboratory and experimental protocols were randomly assigned to either a control (CON, n = 4; 3 ml saline 3 times/week for 3 weeks) or EPO group (EPO, n = 4, 50 iu/kg bwt rhuEPO/3 ml saline 3 times/week for 3 weeks). Exercise tests (GXT) were performed on a treadmill (6% incline), 1 week before and 1 week after treatment. The GXT started at 4 m/sec, with a 1 m/sec increase every 60 sec until the horse reached fatigue. Oxygen uptake was measured via an open flow indirect calorimeter. Blood samples were collected before, during (each step) and 2 and 15 min post GXT to measure packed cell volume (PCV), haemoglobin concentration (Hb), blood lactate concentration (LA) and plasma protein concentration (TP). Plasma volume (PV) was measured using Evans Blue dye. Blood volume (BV) and RCV were calculated using PCV from the 8 m/sec step of the GXT. RESULTS: There were no alterations (P>0.05) in any parameters in CON horses. By week 3, EPO produced increases (P<0.05) in resting PCV (37 +/- 2 vs. 51 +/- 2) and Hb (37%). RCV (26%) and VO2max (19%) increased, but BV did not change (P>0.05) due to decreased PV (-11%, P<0.05). There was a significant increase in velocity at VO2max and LApeak for horses treated with rhuEPO and substantial decrease (P<0.05) in VO2 recovery time when the pretreatment GXT was compared to the post treatment GXT. No differences (P<0.05) were detected for TP, VLA4, run time or Vmax. CONCLUSIONS: Low dose rhuEPO administration increases RCV and aerobic capacity without altering anaerobic power. POTENTIAL RELEVANCE: This study demonstrates that rhuEPO enhances aerobic capacity and exercise performance, a question relevant to racing authorities. PMID- 17402425 TI - Investigation of blood oxidant/antioxidant markers in healthy competition horses of different breeds. AB - REASONS FOR PERFORMING STUDY: There is increasing evidence that the equine athlete is exposed to exercise-induced changes of its oxidant/antioxidant balance and antioxidant supplementation is frequently recommended. However, it is unknown whether there is a specific need for antioxidants according to performance, breed, gender or age. OBJECTIVES: To assess whether breed-, gender- and age related differences of blood oxidant/antioxidant markers occur in competition horses. METHODS: Healthy horses (n = 493) underwent oxidant/ antioxidant blood marker determination. Vitamin E, lipophilic antioxidant capacity (ACL), ascorbic acid (AA), glutathione (GSH, GSSG), gluthione peroxidase (GPx), superoxide dismutase (SOD), selenium (Se), copper (Cu), zinc (Zn), lipid peroxides (Pool), oxidised proteins (Protox) were determined, as well as magnesium (Mg), creatine phosphokinase (CPK), lactate dehydrogenase (LDH), packed cell volume (PCV) and haemoglobin (Hb). A mixed linear model assessed the effect of breed, gender and age category. P<0.05 was considered significant. RESULTS: Thoroughbreds showed the highest values of vitamin E, ACL, GPx, PCV and Hb, whilst standardbreds had the highest values of AA and LDH. Jumping horses had the highest Protox values. Females had significantly higher SOD values, whereas most of the other markers were higher in stallions and geldings. Horses age 2-6 years had higher AA, SOD and LDH values than horses age >6 years. Correlation analyses were positive and significant between vitamin E and GPx, VitE and ACL, Se and GPx, Cu and Pool and negative between Pool and vitamin E, Pool and ACL, Protox and GPx, Protox and vitamin E. CONCLUSIONS: Blood oxidant/ antioxidant status of horses is influenced by breed, gender and age. The correlation analyses suggest synergistic relations between GPx, vitamin E and Se and an antagonistic relation between Protox-GPx, Protox-vitamin E, and Pool-vitamin E. POTENTIAL RELEVANCE: The results of this investigation provide definition of the specific need for antioxidants and vitamins in competition horses. PMID- 17402426 TI - Frusemide results in an extracellular to intracellular fluid shift in horses. AB - REASONS FOR PERFORMING STUDY: Frusemide (Lasix) is commonly used diuretic in horse racing and equine clinical practice. While pharmacology, pharmacodynamics, renal and haematological effects of frusemide have been studied in horses, its effects on the distribution of fluid within the horse remain unknown. OBJECTIVE: To quantify the effects of frusemide on extracellular and intracellular fluid shifts. METHODS: Horses were infused with 1 mg/kg body mass (n = 7) or 2 mg/kg (n = 9) i.v. frusemide. Total body water (TBW), extracellular fluid volume (ECFV) and plasma volume (PV) were measured using D2O, NaSCN and Evans blue dilution. Change in ECFV was assessed from the change in plasma [protein] and from repeated infusion/dilution of NaSCN. RESULTS: Frusemide resulted in a 0.020 +/- 0.002 l/kg decrease in TBW within 120 min. At 120 min after frusemide infusion the ECFV losses were nearly double the TBW losses, therefore ECFV loss in excess of TBW loss is seen as an increase in ICFV. CONCLUSIONS: Frusemide resulted in a net shift of fluid (electrolytes and water) from the extracellular to intracellular fluid compartment. POTENTIAL RELEVANCE: The fluid shifts that occur within horses administered frusemide has not previously been characterised. The intracellular shift of fluid is of performance and clinical significance. PMID- 17402427 TI - Effect of exercise on blood oxidant/antioxidant markers in standardbred horses: comparison between treadmill and race track tests. AB - REASONS FOR PERFORMING STUDY: Interest in establishing oxidant/ antioxidant profiles in competition horses is increasing. Earlier studies performed in horses have mainly been performed under laboratory conditions using a treadmill and it is not known to what extent laboratory results of oxidant/antioxidant studies might be transposed to field conditions. OBJECTIVE: To compare the impact on the blood oxidant/ antioxidant status of a standardised exercise test including a run up to fatigue performed on a treadmill (TM) and on a racetrack (RT) in healthy and trained Standardbred horses. MATERIAL AND METHODS: During TM and RT tests the following blood antioxidant markers were analysed in jugular venous blood at rest and 15 mins (E15) after an intense bout of exercise: uric acid (UA), ascorbic acid (AA), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (reduced: GSH and oxidised: GSSG), glutathione redox ratio (GRR) and protein thiol (PSH). Running time to fatigue (RTF), velocity during the last exercise stage (Vmax), final heart rate (HRfinal) and venous lactic acid (LA) were also recorded. RESULTS: Vmax was significantly (P<0.05) higher during the RT, whereas LA was significantly lower. HRfinal and RTF did not differ significantly between TM and RT. Exercise induced a significant increase (R vs. E15) of UA and AA in both tests, whereas GSH and PSH decreased significantly. GPx, SOD, GSSG and GRR remained unchanged. Differences between TM and RT were significant at E15 for UA, AA and PSH. CONCLUSIONS: Comparison of oxidant/antioxidant profiles from laboratory and field studies are difficult to standardise and should be interpreted with caution. POTENTIAL RELEVANCE: For the same RTF and final HR, the TM induced stronger changes in blood lactate and in blood oxidant/antioxidant balance than did RT. PMID- 17402428 TI - Influence of training on plasma adrenaline and noradrenaline kinetics in untrained standardbreds. AB - REASONS FOR PERFORMING STUDY: Catecholamines (CAT) play an important role in modulating the response to exercise. But the kinetics of CAT changes during exercise are difficult to study due to their short biological half-life. OBJECTIVES: Learning about variations in plasma CAT levels during training could furnish new information regarding sweating, redistribution of blood flow and energy metabolism. METHODS: Four untrained Standardbreds, adapted to treadmill work, were used to determine the influence of training on plasma adrenaline (A) and noradrenaline (NA) kinetics. Horses underwent a standardised exercise test (SET) on treadmill before (SET1), and 1 (SET2) and 2 months after (SET3) the start of a moderate training period on treadmill. The SET procedure was: warm-up and a single step of 2 min at 200 beats/min heart rate (SS). The automatic blood collection system (that has been carried out during SS) and the preparation of the horses have been described previously. The system was programmed to obtain a blood sample every 15 sec (8 blood samples per SS). Analyses were performed by HPLC. Training progress was monitored by means of anaerobic threshold velocity (V4) and the velocity during SS (V200). Wilcoxon's test was used for statistical analysis. RESULTS: Decreased levels of A and NA with significant differences between SET1/SET2 and SET1/SET3 for A (P = 0.007) and NA (P = 0.007) were found. There were no differences between SET2 and SET3 for A (P = 0.195) and NA (P = 0.054). CONCLUSIONS: Training appears to influence the level of plasma CAT and this influence is greater during the first training period in untrained Standardbreds. POTENTIAL RELEVANCE: It may prove interesting to associate the study of training-induced modifications of the concentration of plasma CAT with thermoregulation, which is under catecholaminergic control and undergoes important adaptations during training. PMID- 17402429 TI - The effect of adrenergic suppression induced by guanabenz administration on exercising thoroughbred horses. AB - REASONS FOR PERFORMING STUDY: Adrenergic activity accompanies intense exercise and mediates physiological and metabolic responses to exercise. Guanabenz, an antihypertensive drug marketed for human usage, depresses brain vasomotor and cardioaccelerator centres, blocks peripherally adrenergic neurons and is reportedly used as a calming agent in horses but little is known of its effects in the species. OBJECTIVES: To determine if guanabenz induces measurable signs of adrenergic suppression on fit Thoroughbred horses undergoing intense exercise. METHODS: In a random crossover design, 12 exercise conditioned Thoroughbred horses each received guanabenz (0.08 mg/kg bwt i.v.) and placebo at 3-week intervals. An incremental exercise test to exhaustion on a treadmill followed treatment by 1 h. Heart rate, oxygen consumption, carbon dioxide production, plasma lactate, catecholamines, adrenocorticotropic hormone (ACTH) and cortisol, and time to fatigue were monitored. Statistical analysis was performed using mixed-effects linear modelling. RESULTS: Mean heart rate during the exercise period was lower in guanabenz-treated horses (P = 0.04). Mean concentrations of plasma cortisol (P = 0.02) and adrenaline (P = 0.03) were lower for guanabenz treated horses during the exercise period. Mean run time was slightly but not significantly longer for guanabenz-treated horses, (P = 0.053). No significant effects of guanabenz administration were found for oxygen consumption, carbon dioxide production nor for plasma lactate, noradrenaline and ACTH concentrations. CONCLUSION: Guanabenz administration induced signs of adrenergic suppression including plasma cortisol and adrenaline concentrations and heart rate and may enhance endurance, but did not eliminate increases in hormone concentrations induced by exercise. Clear determination of a positive performance effect of adrenaline, but not noradrenaline, suppression is needed before clinical significance can be determined. PMID- 17402430 TI - Plasma beta-endorphin, cortisol and immune responses to acute exercise are altered by age and exercise training in horses. AB - REASONS FOR PERFORMING STUDY: Ageing appears to affect immune and neuroendocirne function in horses and response to acute exercise. No studies have examined the combined effects of training and ageing on immune and neuroendocirne function in horses. HYPOTHESIS: To ascertain whether training and age would affect the plasma beta-endorphin (BE) and cortisol (C) as well as immune function responses to acute exercise in Standardbred mares. METHODS: Graded exercise tests (GXT) and simulated race tests (SRT) were performed before and after 12 weeks training at 60 % HRmax. BE and C were measured at rest and at 5, 10, 20, 40, 60 and 120 min post GXT. Leucocyte cell number, CD4+ and CD8+ lymphocyte subsets, and mitogen stimulated lymphoproliferative response (LPR), were measured in jugular blood before and after the SRTs. RESULTS: Cortisol rose by 5 min post GXT in young (Y) and middle-age (MA) mares (P<0.05) and remained elevated until 40 and 60 min post GXT, respectively during both pre- and post training GXT. There was no rise in C in old (0) mares after either GXT (P>0.05). Pretraining BE rose (P<0.05) by 5 min post GXT in all mares. After training, BE was higher in Y and O vs. MA (P<0.05) at 5 min post GXT. Post training BE was higher at 5 min post GXT in Y and O vs. pretraining (P<0.05). After SRT, lymphocyte number rose in all mares (P<0.05); however, lower lymphocyte numbers (P<0.05) were seen in MA vs. Y and O vs. MA (P<0.05). The O had reduced LPR to Con A and PHA stimulation (P<0.05) compared to Y and MA after the SRT after both pre- and post training SRT. LPR to PWM was lower (P<0.05) in O vs. Y and MA after the pretraining SRT. Training caused an increase in resting LPR to PWM in MA only (P<0.05). CONCLUSION: Both age and training altered the plasma beta-endorphin and cortisol responses as well as and immune responses to acute exercise. POTENTIAL RELEVANCE: This study provides important information on the effects of ageing and training that will aid in the management and care of an increasing number of active older horses. PMID- 17402431 TI - Hormone response to training and competition in athletic horses. AB - REASONS FOR PERFORMING STUDY: It is recognised that the amount of psychological stress that an animal encounters determines the degree of response of the hypothalamic-pituitary-adrenal (HPA) axis. In human athletes, the added emotive stress of competition is an important element in the adrenal response. The aim of this study was to examine the effect of show-jumping as well as dressage on stress levels by comparing horses' stress response at a horse show compared to their familiar home. METHODS: Fifty-one horses involved in competition were used. EDTA blood samples were collected before exercise, upon arrived to the schooling area (control), and k over a jump or dressage course. After sampling, plasma was separated and stored at -80 degrees C until determinations of cortisol and ACTH were performed. Fourteen healthy horses not involved in competition were used as control group. RESULTS: Competition induced a significant increase in cortisol and ACTH responses in both, jumping and dressage horses and this effect was more apparent in dressage horses. When horses were most experienced, cortisol and ACTH responses were much lower. CONCLUSION: This study shows that competition elicits a classic physiological stress response in horses and that different training programmes induce different responses. It suggests that horses involved in competition can provide a good model to study the exercise-induced stress response. PMID- 17402432 TI - The effects of dietary N-3 and antioxidant supplementation on erythrocyte membrane fatty acid composition and fluidity in exercising horses. AB - REASONS FOR PERFORMING STUDY: Fatty acid supplementation could modulate erythrocyte membrane fluidity in horses at rest and during exercise, but information is lacking on the effect of exercise. OBJECTIVES: To assess the effect of exercise with, and without, an oral antioxidant supplementation enriched with n-3 fatty acids on erythrocyte membrane fluidity (EMF) and fatty acid composition in eventing horses. METHODS: Twelve healthy and regularly trained horses were divided randomly into 2 groups: group S received an oral antioxidant cocktail enriched in n-3 fatty acid (alphatocopherol, eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) whereas group P was placebo-treated. At the end of 4 weeks, all horses performed a standardised exercise test (ET) under field conditions. Venous blood was sampled before starting treatment (TO), immediately before (T1) as well as 15 min (T2) and 24 h (T3) after ET. Spin labelled (16-DOXYL-stearic acid) red blood cell membranes were characterised using the relaxation correlation time (Tc in inverse proportion to EMF). Fatty acid composition (%) of the membrane was determined by gas-liquid chromatography. RESULTS: Supplementation did not induce changes in EMF (T1 vs. TO) but significant changes in membrane composition were observed and there were increases in n-3 polyunsaturated fatty acid PUFA, n-3/n-6 ratio, and total n-3 fatty acids. Exercise (T2 vs. T1) induced a significant decrease of EMF in group P (Tc: +19%, P<0.05) and nonsignificant decrease in group S (Tc: +5%), whereas membrane fatty acid composition did not change in either group. During the recovery period (T3 vs. T2), EMF decreased significantly in group S (Tc: +29%, P<0.05) and nonsignificantly in group P (Tc: +18%) without any significant changes in fatty acid composition. CONCLUSION AND POTENTIAL RELEVANCE: An enriched oral antioxidant supplementation induced changes in membrane composition, which modulated the decrease in EMF induced by exercise. Long chain n-3 fatty acid supplementation might therefore be beneficial. PMID- 17402433 TI - Hydration of exercised standardbred racehorses assessed noninvasively using multi frequency bioelectrical impedance analysis. AB - REASONS FOR PERFORMING STUDY: In human and animal clinical practice, multi frequency bioelectrical impedance analysis (MF-BIA) is increasingly used as a diagnostic tool to assess hydration of intra-and extracellular fluid compartments. Accurate determination of changes in hydration status within individuals over time has remained problematic due to the requirement for complete impedance-frequency relationships at the time points of interest. OBJECTIVES: To use MF-BIA in 13 Standardbred racehorses and 7 'endurance' research horses to determine if MF-BIA could be used to track changes in total body water (TBW), intracellular fluid volume (ICFV) and extracellular fluid volume (ECFV) resulting from exercise. METHODS: Jugular venous blood was sampled at rest and for 2-13 h following exercise. TBW, ECFV and plasma volume (PV) were measured at rest using indicator dilution techniques (D2O, thiocyanate and Evans Blue, respectively). TBW, ECFV, ICFV and PV were correlated to impedance measures and predictive equations used to determine hydration status from MF-BIA measures. RESULTS: TBW loss continued throughout the recovery period, and was primarily borne by the ECF compartment at 90 min of recovery. CONCLUSIONS: MF-BIA predictions of compartmental hydration status were significantly correlated to measured/calculated decreases in these compartments. POTENTIAL RELEVANCE: Practical applications for MF-BIA in horses include monitoring of hydration status during transport and competition, assessment of body compostion, clinical health assessment and critical care management. PMID- 17402434 TI - Is improved high speed performance following frusemide administration due to diuresis-induced weight loss or reduced severity of exercise-induced pulmonary haemorrhage? AB - REASONS FOR PERFORMING STUDY: Prerace administration of frusemide to horses has been linked with a significant improvement in racing performance, but the basis for this improvement is unclear. OBJECTIVE: To test whether improved performance with prerace administration of frusemide is due to the drug's diuresis-induced weight loss rather than its apparent alleviation of exercise-induced pulmonary haemorrhage (EIPH). METHODS: Eight thoroughbred horses underwent 3 trials in a random order, 2 or 3 weeks apart: control (C), frusemide/unburdened (FU), and frusemide/burdened (FB). None of the horses were known to have exhibited post exercise epistaxis or endoscopic evidence of EIPH. Endoscope-guided bronchoalveolar lavages (BALs) were performed before and after each horse completed a standardised exercise test (SET) on an inclined treadmill to assess semi-quantitatively the volume of EIPH. For C, horses received an i.v. saline placebo injection (5 ml) and were unburdened while performing the SET. With FU, horses received frusemide (0.5 mg/kg) and were also unburdened. For FB, horses received frusemide and were burdened with weight equal to that lost during the 4 h post frusemide injection period. Erythrocyte number in BAL fluid, mass specific VO2max, time and distance for the entire SET as well as at maximum speed were recorded. A one-way repeated measures analysis of variance was conducted on all results. RESULTS: Mass specific VO2max was significantly higher for the FU than for FB or C. Mass specific VO2max for FB and C were not different. More RBCs were found in BAL samples after C runs than after both FU and FB trial runs. Horses with the frusemide treatment (either burdened or unburdened) produced less EIPH than in the C trial, but their mass specific VO2max values were higher on the FU trial alone. For FU, horses ran longer at 115% VO2max than under C or FB conditions. CONCLUSION AND POTENTIAL RELEVANCE: Improvement of performance in the furosemide trials was due more to the weight-loss related effects of the drug than its apparent alleviation of EIPH. Further research is warranted with the same or similar project design, but with a larger sample size and with horses known to have more severe EIPH. PMID- 17402435 TI - Changes in arterial, mixed venous and intraerythrocytic concentrations of ions in supramaximally exercising horses. AB - REASONS FOR PERFORMING STUDY: Horses experience major perturbations in acid-base balance during supramaximal exercise. Ion movement in and out of erythrocytes (RBCs) is believed to be important in maintaining acid-base balance but it is unclear as to the extent to which this happens, nor how it affects single measurements of ion concentrations in arterial and venous blood. OBJECTIVES: To clarify the role RBCs play in mitigating perturbations in acid-base balance during high speed exercise in horses, and to describe associated differences in arterial (a) and mixed venous (v) concentrations of key ions. METHODS: Six exercise-trained Thoroughbreds galloped to fatigue at speeds calculated to have an oxygen demand that was 115% of the VO2max. Blood samples (a and v) were collected pre-exercise, during warm-up, at fatigue, and immediately post exercise. Packed cell volume (PCV), pH, PCO2, and plasma concentrations of bicarbonate (HCOP3-), chloride (Cl-), sodium (Na+), potassium (K+), and lactate (Lac-) and strong ion difference (SID) were determined, and RBC concentrations of Lac- and electrolytes calculated for each sample. Data were analysed using a 2 way ANOVA for repeated measures testing for effects of sampling time and site (P<0.05). RESULTS: Plasma and RBC [Cl-] were increased with hypercapnoea and acidaemia. [HCO3-]v was greater than pre-exercise values at fatigue, although [HCO3l]a was lower. Hyperkalaemia and decreased RBC [K+] were evident at fatigue, as was an increased RBC [Na+]. Plasma [K+] started to decrease as soon as exercise ceased and Na+ began to move back onto RBCs in exchange for K+. Concentrations of all measures of Lac- rose from fatigue to post exercise. The SID decreased with exercise and was higher in v at fatigue and post exercise, reflecting the decrease in pH. CONCLUSIONS: RBCs act as a repository for lactate, and therefore the increase in PCV facilitates the maintenance of the muscle to plasma Lac- diffusion gradient during exercise. POTENTIAL RELEVANCE: This serves to keep intramuscular [Lac-] lower than it would otherwise be and, because of the link between Lac- accumulation, pH decrease and the onset of fatigue, may help delay the onset of fatigue. PMID- 17402436 TI - Plasma concentrations of myeloperoxidase in endurance and 3-day event horses after a competition. AB - REASON FOR PERFORMING STUDY: In man, exercise of any type has been shown to induce neutrophil degranulation and respiratory burst activity, as well as an increase in plasma myeloperoxidase (MPO), a specific enzyme of neutrophil azurophilic granules with a strong oxidative activity. Until now, it is not known whether this is the same in horses. OBJECTIVES: To study whether degranulation of blood neutrophils may be induced by exercise by measuring the total concentration of MPO and assess the possible influence of type of competition on this exercise induced adjustment. METHODS: Blood was sampled before, and 30 min after, the course, in 9 ponies performing the Phase D of a national 3-day event championship (CIC*), and in 7 endurance horses participating at the European endurance championship 2005. White blood cells and granulocytes, total plasma proteins, creatine phosphokinase (CK), and total MPO contents were determined from blood samples. In addition, blood was taken from all ponies and 4 of the endurance horses 5 min after completion of the course to give some idea of the intensity of exercise. RESULTS: The mean blood lactate was 15.8 +/- 5.8 mmol/l after the CIC* and 2.7 +/- 0.2 mmol/l after the 160 km course. Performing both competitions induced a significant increase in CK and MPO. After the endurance course, the number of granulocytes significantly increased. Whilst there was no significant correlation between the measurements in CIC* ponies, MPO was significantly correlated with granulocyte count (r2 = 0.776) and CK (r2 = 0.586) in endurance horses. CONCLUSIONS: Intense exercise induces an activation of blood granulocytes, with degranulation of neutrophils and release of MPO. The plasmatic MPO concentration after endurance was higher than the values reported in some inflammatory pathological conditions. POTENTIAL RELEVANCE: This phenomenon may partly contribute to the occurrence of an exercise-induced oxidative stress and to the alteration of muscular membrane permeability. Further studies should be conducted to assess the possible relationship between MPO concentration and markers of oxidative stress in performance horses. PMID- 17402437 TI - Effects of a pre-moistened multilayered breathable fabric in promoting heat loss during recovery after exercise under hot conditions. AB - REASONS FOR PERFORMING STUDY: Horses generate considerable internal heat burdens when exercising. Although common practice for a trainer or groom to place a wet blanket or towel on the dorsum of a hot horse post exercise, there are no data supporting the efficacy of this cooling method. OBJECTIVE: To test the hypothesis that a pre-moistened blanket designed with a multilayered breathable fabric would enhance heat loss in horses post exercise. METHODS: Eight treadmill-trained horses performed a standardised exercise test (SET) weekly for 3 weeks, with 3 different recovery treatments administered randomly. Pulmonary artery temperature (PAT) was measured via Swan-Ganz catheter. The SET consisted of 10 min at 3.7 m/sec, 3 min at 11.0 m/sec, 25 min at 3.7 m/sec and 20 min of recovery walking at 2.0 m/sec (58 min exercise and recovery under laboratory conditions of 35.0-40.6 degrees C and 27-49% RH). From 3-7 min during recovery, the treadmill was stopped and horses randomly received either: (a) no bath (negative control); (b) a bath consisting of 32 l of 1-4 degrees C water split into 3-4 cycles of bilateral water application (positive control) followed by water removal ('scraping'); or (c) application of a multilayered fabric blanket soaked in 16-19 degrees C water, wrung out, and placed over the dorsum and sides of the horse. PAT was compared using RM ANOVA with the Student Neuman-Keul's test used post hoc to discriminate between treatments at specific points in time. RESULTS: Mean PAT rose with each phase of exercise (P<0.001) and peaked at a mean of 40.2 +/- 0.2 degrees C. During recovery, the cold bath decreased HR and PAT for 9 min after walking resumed (P<0.001-P<0.05). The blanket did not decrease HR or PAT compared to negative control (P>0.05), and both were hotter than the cold bath treatment through 16 min of recovery (P<0.05). CONCLUSIONS: A specially-designed cooling blanket failed to reduce PAT when compared to negative control. Cold water bathing decreased HR and PAT but was not effective throughout all of recovery. POTENTIAL RELEVANCE: A specially-designed, pre-moistened multilayered breathable fabric failed to promote evaporative cooling compared to negative control. Cold water baths may need to be repeated throughout recovery to optimise their effect. PMID- 17402438 TI - Contribution of exercise intensity and duration to training-linked myosin transitions in thoroughbreds. AB - REASONS FOR PERFORMING STUDY: The influence of relevant exercise parameters such as intensity and duration of exercise on the muscular response to training has not been reported in Thoroughbreds. This knowledge may be of great value in practice. OBJECTIVE: To define the relative contribution of the intensity and duration of exercise on the myosin heavy chain (MHC) response to training in Thoroughbred racehorses. METHODS: Six 2-3-year-old Thoroughbreds, following a randomised 6 x 6 latin square design, performed 6 consecutive conditioning programmes of varying lactate-guided intensities and durations. Each programme lasted 22 days and consisted of 11 exercise sessions once a day, every second day on a 6% inclined treadmill followed by a 10 day resting period between consecutive programmes. Gluteus muscle biopsies (depth, 2 and 6 cm) were taken before, and after, each programme and electrophoretically analysed for MHCs. RESULTS: Overall, training increased the fraction of MHC IIA (mean +/- s.d., 41.1 +/- 3.5% to 43.4 +/- 4.42% P<0.001) and decreased that of MHC IIX isoform (from 50.2 +/- 4.4% to 47.4 +/- 5.6% P<0.001). Exercise intensity fixed effects on fast MHCs were highly significant (P<0.01), whereas exercise duration only had a marginal effect (P<0.1) on the IIA:IIX MHC ratio. On a per-individual basis, training impact on fast MHCs was only significant (P<0.05) in horses exercised for the longest duration at both intensities. CONCLUSION: The short-term training induced up-regulation of MHC IIA and down-regulation of MHC IIX in Thoroughbreds are more dependent on intensity than duration of exercise. However, protocols with exercises of moderate intensity and long duration can induce MHC changes similar to those promoted by programmes with exercises of higher intensities. POTENTIAL RELEVANCE: Exercises for up to 25 min/day at velocities eliciting a blood lactate concentration between 2.5 and 4 mmol/l every second day for 3 weeks improve stamina in Thoroughbreds. PMID- 17402439 TI - Myosin heavy chain fibre type composition in foals: analyses at the mRNA and protein level. AB - REASONS FOR PERFORMING STUDY: An optimal developed musculoskeletal system is vital for the performance of the horse. Previously, we showed that in the m. gluteus medius from adult untrained horses, identical mRNA and protein expression patterns were found in the majority of fibres. However, co-expression of IIa and IId/x myosin heavy chain (MyHC) was substantially more common at the protein than at the mRNA level, suggesting a transcriptionally controlled fine-tuning of these 2 genes. OBJECTIVE: To analyse the MyHC transcripts and proteins (including the cardiac alpha isoform) in the same muscle during post natal development when the muscle is adapting to movement and load. METHODS: Biopsies were taken from the m. gluteus medius of 2 Dutch Warmblood foals at 0, 2, 4, 22 and 48 weeks of age. mRNA was compared to protein expression on a fibre-to-fibre basis using in situ hybridisation and immunofluorescence. The MyHC slow (I), alpha, IIa and IId/x isoforms were analysed. RESULTS: At all ages the expression of the mRNA and protein MyHC isoforms was almost identical. Surprisingly, coexpression of the IIad isoform was also detected at the mRNA level especially early in life. The transcript of the alpha isoform was only detectable at young age, indicating silencing of the gene around birth. CONCLUSION: During the first year of life, MyHCs are continuously adapting at the mRNA and protein level. Additionally, the regulation of hybrid fibres is different from that in adult fibres. POTENTIAL RELEVANCE: We postulate that interfering in this process by e.g. early training will be levelled out by the maturation of the muscle. PMID- 17402440 TI - MCT1 and CD147 gene polymorphisms in standardbred horses. AB - REASONS FOR PERFORMING STUDY: Transport of lactate across membranes is facilitated by proton-monocarboxylate transporters (MCT). The most widely distributed isoform is MCT1, which needs an ancillary protein CD147. Studies on erythrocytes have shown that high activity of MCT1 is inherited as the dominant allele and that activity is regulated through CD147. Mutations of human MCT1 have been described that appear to impair lactate transport in muscles and cause exertional rhabdomyolysis. There are no reports of this potential relationship in the horse. OBJECTIVES: To obtain sequences of equine MCT1 and CD147 to examine differences between horses with high and low lactate transport activity in their erythrocytes. METHODS: Muscle biopsy samples were taken from 3 healthy Standardbred horses and from 7 horses which according to the owners had signs of myopathy after intense exercise. DNA and RNA were isolated and PCR analysis and sequencing performed. RESULTS: Currently, PCR fragments covering 100% of MCT1 and 70% of CD147 coding region are retained and sequence analysis has demonstrated one single nucleotide polymorphism (SNP) in the C-terminal area of MCT1 and one SNP in the extracellular domain of CD147. Both cause an amino acid change. The SNPs found are not related to lactate transport activity in erythrocytes or signs of myopathy. CONCLUSIONS: More samples need to be analysed to make conclusions on the significance of the polymorphisms found. Furthermore, full sequence coverage of the coding region of CD147 is needed. POTENTIAL RELEVANCE: The molecular probes produced could be used as tools to study gene regulation of lactate transport. PMID- 17402441 TI - Muscle characteristics of dutch warmblood foals with different genetic background at ages 6 and 12 months. AB - REASONS OF PERFORMING STUDY: To obtain broader insight into the muscle of foals with different genetic background, muscle fibre composition, its post natal development and citrate synthase (CS) activity of the gluteus medius were investigated. HYPOTHESIS: Because muscle properties are influenced genetically and related directly to performace ability, muscle characteristics should be affected by selection and correspond with the requirement of ths sports selected for. METHODS: The foals were divided into Group A, (n = 16), considered an average of the population, and Group B (n = 36), selected for showjumping. Biopsies were taken deeply from the gluteus medius muscle. Fibre type identification was performed immunohistochemically at ages 6 and 12 months. Citrate synthase activity was measured at age 6 months. RESULTS: age 6 months statistically significant differences were found between Groups A and B in the proportion of type I, Had and IId fibres. Oxidative fibres (I and IIad) were more abundant in Group B, while fast-glycolytic fibres (IId) were more abundant in Group A. Except for IIa, the fibre type composition at age 12 months had become equal in the 2 groups. The difference in the sum of oxidative fibres (I + IIad + IIad) between Groups A and B at age 6 months was similar to the difference in CS activity at that age, although the latter was not significant. CONCLUSIONS: At age 6 months the 2 groups had different proportions of fibre types I and IId, but these differences had disappeared at age 12 months. POTENTIAL RELEVANCE: The muscular response to training at an early age can be interpreted with knowledge of muscle characteristics and its post natal development in foals selected for showjumping. PMID- 17402442 TI - Effects of prior exercise on components of insulin signalling in equine skeletal muscle. AB - REASONS FOR PERFORMING STUDY: Exercise is an important stimulus for the regulation of numerous metabolic functions in muscle, but there has been little investigation of the mechanism of exercise-enhanced glucose utilisation. OBJECTIVE: To examine the effects of exercise on the signalling components Akt and glycogen synthase kinase-3 (GSK3) involved in insulin regulation of glycogen synthase (GS) activity. It was hypothesised that hyperinsulinaemia and prior exercise would alter the extent of Akt phosphorylation and GSK3beta deactivation and increase insulin stimulation of GS activity in skeletal muscle. METHODS: Eight Standardbred horses completed a euglycaemic-hyperinsulinaemic clamp (EHC) either without prior exercise (NonEX) or at 0.5, 4 or 24 h post exercise (0.5 PostEX, 4 PostEX, and 24 PostEX, respectively). Muscle samples for determination of Akt and GSK3 serine phosphorylation, and GS fractional velocity (GS(FV)) were collected 5 min before (Pre-INS) and at the end (Post-INS) of each EHC (pre- and post insulin stimulation). Protein was separated by SDS-PAGE on polyacrylamide gels, transferred to nitrocellulose membranes and incubated with antibodies against phospho-Akt Ser473 and GSK3alpha/beta Ser21/9. RESULTS: GS(FV) was increased (P<0.05) 2-fold at 0.5 PostEX, 4 PostEX and 24 PostEX when compared to NonEX in Pre-INS and in Post-INS was increased 33 and 28% in NonEX and at 24PostEX (P<0.05), respectively, when compared to Pre-INS in these trials. Phospho-Akt Ser473 was increased at 0.5 PostEX only (P<0.05) in Pre-INS but increased in all trials in Post-INS. Phospho-GSK3alpha (Ser21) was also greater (P<0.05) at 0.5 PostEX in Pre-INS than in NonEX and 24 PostEX and in Post-INS increased (P<0.05) in NonEX and at 24 PostEX but not at 0.5 PostEX or 4 PostEX post exercise. There was no effect of time or treatment on GSK3, (Ser9) phosphorylation or on total GSK3. CONCLUSIONS: Elevated plasma insulin and prior exercise were associated with Akt activation and GSK3alpha deactivation (0.5 PostEX) indicating that an effect of exercise is evident in the early post exercise period and supporting a physiological role for GSK3alpha in the regulation of GS activity. The changes are consistent with insulin signalling downstream from these molecules. PMID- 17402443 TI - Synthesis of proglycogen and macroglycogen in skeletal muscle of standardbred trotters after intermittent exercise. AB - REASONS FOR PERFORMING STUDY: The degradation of glycogen and its two forms, proglycogen (PG) and macroglycogen (MG) has been studied in horses performing different types of exercise, but no information is available about the resynthesis of PG and MG after exercise. OBJECTIVES: To determine the resynthesis of PG and MG in skeletal muscle after intermittent uphill exercise. METHODS: At a training camp 9 well-trained Standardbred trotters performed a training session comprising a warm-up period, 7 repeated 500 m bouts of exercise on an uphill slope and a recovery period. Muscle biopsies (m. gluteus medius) for analysis of PG, MG, glucose and glucose-6-phosphate were taken at rest, at the end of exercise and 1, 4, 8, 24, 48 and 72 h post exercise. Blood samples for analysis of glucose, lactate and insulin were collected before exercise, immediately after the last bout of exercise and then as for the muscle biopsies. RESULTS: The MG and PG concentration pre-exercise was 311 - 47 and 305 +/- 55 mmol/kg dwt respectively. The exercise caused a decrease in PG (A 63 +/- 26 mmol/kg dwt) and MG (delta 136 +/- 68 mmol/kg dwt). Immediately after the last sprint plasma glucose and lactate increased compared to values pre-exercise. During the first hour post exercise there was a further decrease in MG in 7 out of 9 horses. The rate of glycogen resynthesis during 1-24 h was higher for MG than for PG. The rate of muscle glycogen resynthesis thereafter was slower and did not differ between MG and PG up to 72 h. CONCLUSION: After repeated bouts of exercise on a slope, resynthesis of glycogen is a slow process and the resynthesis of proglycogen differs from that of macroglycogen. The fraction most depleted during exercise (MG) had no resynthesis during the first hour of recovery but then had the highest rate of resynthesis during the remainder of the first 24 h period. POTENTIAL RELEVANCE: If the time between exercise sessions during training is too short the recovery period will be inadequate for complete restoration of muscle glycogen. PMID- 17402444 TI - Prevalence of polysaccharide storage myopathy in horses with neuromuscular disorders. AB - REASONS FOR PERFORMING STUDY: Controversy exists as to the prevalence of polysaccharide storage myopathy (PSSM) in breeds of horses and its impact on performance. OBJECTIVES: To determine 1) the prevalence of PSSM in horses that presented with a neuromuscular disorder, as well as breed, sex and age distributions and clinical signs 2) effect of diagnostic criteria on prevalence, breed distribution and age of horses diagnosed with PSSM. METHODS: Fresh frozen biopsies (n = 1426) submitted to the Neuromuscular Diagnostic Laboratory at the University of Minnesota were searched to identify horses diagnosed with PSSM. Horses with and without histological evidence of PSSM were compared. Biopsies were classified as Grade 1, containing aggregates of granular glycogen or Grade 2, containing periodic acid Schiff's (PAS) positive inclusions, traditionally resistant to amylase digestion. RESULTS: Horses (n = 572 : 40.1%) were identified with PSSM, of which 62.9% were Quarter Horse related breeds (QHR), 11.5% Draught breeds (DB) and 8.9% Warmblood breeds (WB). Exertional rhabdomyolysis was more prevalent in QHR than DB and WB, whereas QHR were less likely to have muscle atrophy compared to DB. QHR were less likely to have gait abnormalities than DB and WB. The highest within breed prevalence of PSSM was in DB at 63/116, WB 58/111 and QHR 360/753. Exclusion of Grade 1 criteria decreased the overall prevalence of PSSM to 21.7% of biopsy submissions, and decreased the within breed prevalence in each breed category. The within breed prevalence decreased most substantially in the breeds less commonly diagnosed with PSSM, Thoroughbreds (4.5%) and Arabians (2.5%). CONCLUSION: PSSM is a common cause of neuromuscular disease in QHR, DB and WB related breeds. Inclusion of granular glycogen as the sole diagnostic criterion may increase the sensitivity of this diagnostic test, but conversely it may decrease the specificity of the diagnosis resulting in the inclusion of horses of Thoroughbred, Arabian and other breeds. POTENTIAL RELEVANCE: PSSM is an important differential diagnosis for QHR, WB and DB presenting with signs of rhabdomyolysis, gait abnormalities and muscle atrophy. PMID- 17402445 TI - Muscle, tendon, and somatotropin responses to the restriction of muscle blood flow induced by KAATSU-walk training. AB - OBJECTIVE: The efficacy of KAATSU training has been demonstrated in human athletes, both as a therapeutic method as well as a training aid. The purpose of this study was to investigate the effects of slow walk training combined with restriction of muscle blood flow (KAATSU) on muscle and tendon size. METHODS: Six healthy, unfit Standardbred mares performed walking (240 m/min for 10 min and then 5 min recovery) with KAATSU, and 6 mares performed walking without KAATSU. A specially designed elastic cuff1 was placed at the most proximal position of the forelegs and inflated to a pressure of 200-230 mmHg throughout the walking and recovery sessions. The training was conducted once a day, 6 days/week for 2 weeks. Skeletal muscle thickness and tendon thickness were measured using B-mode ultrasound at baseline and after 2 weeks of training. Venous blood samples were obtained before the first acute exercise and 5, 15 and 60 min afterwards. Serum somatotropin concentration was determined using a commercially available equine specific ELISA kit. RESULTS: The acute increase in plasma somatotropin was 40% greater (P<0.05) in the KAATSU-walk group than in the Control-walk group 5 min after exercise and remained elevated (P<0.05) at 15 and 60 min post exercise compared with the Control-walk group. After 2 weeks of training, muscle thickness increased (P<0.05) 3.5% in the KAATSU-walk group but did not change in the Control-walk group (0.7%). Tendon thickness did not change (P>0.05) in either group. CONCLUSIONS: These data demonstrate that KAATSU training can induce muscle hypertrophy in horses and suggest that KAATSU training may provide significant therapeutic/ rehabilitative value in horses, as has been shown in man. PMID- 17402446 TI - Recruitment pattern of muscle fibre type during flat and sloped treadmill running in thoroughbred horses. AB - REASONS FOR PERFORMING THE STUDY: There is little information about the muscle fibre recruitment pattern during sloped and flat track running in Thoroughbred horses. OBJECTIVES: To examine the glycogen depletion pattern of each muscle fibre type during running on a flat and sloped treadmill. METHODS: Thirteen Thoroughbred horses (3-9 years old) were used. They were initially subjected to incremental exercise tests on a treadmill at 10 and 0% inclines in each horse to determine running speed at 90 and 60% VO2max. Needle biopsy samples were obtained from the middle gluteal muscle immediately after the running at 90% VO2max for 4 min and 60% VO2max for 12 min on 10% and 0% inclines treadmill. Four muscle fibre types (Types I, IIA, IIA/IIX, and IIX) were immunohistochemically identified, and optical density of Periodic Acid Schiff staining (OD-PAS) in each fibre type and the glycogen content of the muscle sample were determined by quantitative histochemical and biochemical procedures. RESULTS: The changes in OD-PAS showed that the recruitment of all fibre types were identical after each exercise bout, i.e., 4 min running at 90% VO2max (8.4-9.4 m/sec on 10%, 13.9-14.1 m/sec on 0%), and 12 min running at 60% VO2max (5.4-6.0 m/sec on 10%, 7.9-11.2 m/sec on 0%). No significant differences were found in the recruitment patterns of each muscle fibre type between 10 and 0% inclined exercise bouts at the same exercise intensity. CONCLUSIONS: The recruitment pattern of muscle fibre type is mainly determined by exercise intensity (%VO2max) and duration, but not by running speed. POTENTIAL RELEVANCE: The results of this study indicate the possibility that up-hill running results in the same training effect as faster running on a flat track. PMID- 17402447 TI - Monocarboxylate transporters (MCT) as lactate carriers in equine muscle and red blood cells. AB - REASONS FOR PERFORMING STUDY: Monocarboxylate transporters (MCT) facilitate the transport of lactate across membranes. In red blood cells (RBC) the transport activity varies interindividually due to differences in the amount of an ancillary protein CD147. Similar variations in muscles could have a great influence on lactate accumulation during exercise. OBJECTIVES: To study the expression of MCT isoforms and CD147 in the middle gluteal muscle. METHODS: Venous blood and muscle biopsy samples were taken from 14 Standardbred horses. Lactate transport activity in RBC and the amounts of MCT1, 2, 4 and CD147 were measured. RESULTS: In muscle MCT1, MCT4 and CD147 were found. Amount of MCT1 was variable and not dependent on age or training. Expression of MCT4 increased with age and correlated positively with CD147. CD147 in muscle correlated with that in RBC. MCT4 in muscle and CD147/MCT1 in RBC were higher in race fit than in moderately trained horses. CONCLUSIONS: MCT isoform profile in equine muscle is similar to that in man. The correlation between CD147 in muscle and RBC supports the view that lactate transport activity in muscles may vary interindividually as with RBC. POTENTIAL RELEVANCE: A larger number of horses need to be analysed to confirm the relationship of CD147 in muscle and RBC; and to allow the use the lactate transport activity in RBC as an indicator of the respective activity in muscles. PMID- 17402448 TI - Gene expression profiling in equine muscle tissues using mouse cDNA microarrays. AB - REASONS FOR PERFORMING STUDY: Progress could be achieved by using microarrays to understand metabolic adaptations and disorders in equine muscle in response to exercise. OBJECTIVES: To test the feasibility of using mouse cDNA microarrays to analyse gene expression profile in normal equine muscles. METHODS: Muscular biopsies of dorsal gluteus medius and longissimus lumborum were done in 4 healthy Standardbreds. Total RNA was extracted from the muscle samples. The concentration and quality of RNA were measured before and after amplification. Gene expression profiles were measured using mouse cDNA microarrays including 15,264 unique genes representing about 11,000 documented genes. Three hybridisation tests were performed to check interspecificity, reproducibility and to compare gene expression in these muscles. For each test, a dye-swap hybridisation with Cy3 and Cy5 fluoromarkers were done and the gene list filtered according the signal level. RESULTS: According to the specificity test, the mouse cDNA microarrays were correctly hybridised by equine muscle cDNA. All positive control genes (GAPDH, HPRT and beta-Actin) and no negative control gene (yeast, plant) hybridised. The reproducibility test demonstrated a good linearity between the duplicate hybridisations: 99.99% of the significant expressed genes have an expression ratio between 1.4 and 1/1.4 = 0.71. These limits can be considered as the thresholds to qualify as up-regulated (ratio >1.4) or downregulated (ratio <0.71). In the muscle comparison test between gluteus medius vs. longissimus lumborum, 63 genes were found up-regulated and 8 genes down-regulated. The range of gene expression ratios in the gluteus medius was 0.61-8.31 x the longissimus lumborum. This list of modulated genes was classified by functions using a gene ontology data basis. CONCLUSION: Mouse microarrays could be used to hybridise equine RNA extracted from muscle tissues. For many genes there are large sequence identities that allowed interspecific cDNA hybridisation. The sensitivity of the method allowed quantification of up- and down-regulated genes after applying appropriate filters. POTENTIAL RELEVANCE: Expression profiling could be used to explore the muscle metabolism changes related to exercise, training, pathology and illegal medication in horses. PMID- 17402449 TI - Energetic cost of locomotion on different equine treadmills. AB - REASON FOR PERFORMING STUDY: Human athletes run faster and experience fewer injuries when running on surfaces with a stiffness 'tuned' to their bodies. We questioned if the same might be true for horses, and if so, would running on surfaces of different stiffness cause a measurable change in the amount of energy required to move at a given speed? HYPOTHESIS: Different brands of commercial treadmills have pans of unequal stiffness, and this difference would result in different metabolic power requirements to locomote at a given speed. METHODS: We tested for differences in stiffness between a Mustang 2200 and a Sato I commercial treadmill by incrementally loading each treadmill near the centre of the pan with fixed weights and measuring the displacement of the pan as weights were added or removed from the pan. We trained six 3-year-old Thoroughbreds to run on the 2 treadmills. After 4 months the horses ran with reproducible specific maximum rates of O2 consumption (VO2max/kg bwt, 2.62 +/- 0.23 (s.d.) mlO2 STPD/sec/kg) at 14.2 +/- 0.7 (s.d.) m/sec. They were alternately run on the 2 treadmills at identical grade (0.40 +/- 0.02%) and speeds (1.83 (walk), 4.0 (trot) and 8.0 (canter) m/sec, all +/- 0.03 m/sec) while wearing an open-flow mask for measurement of VO2. RESULTS: The Mustang treadmill was over 6 times stiffer than the Sato. The VO2/kg bwt increased by approximately 4-fold over the range of speeds studied on both treadmills. Oxygen consumption was significantly lower at all speeds for the Mustang treadmill compared to the Sato. The fractional difference in energy cost decreased by a factor of 6 with increasing speed, although absolute difference in cost was relatively constant. CONCLUSIONS: We suggest it costs less energy for horses to walk, trot or canter on a stiffer treadmill than on a more compliant treadmill, at least within the ranges of stiffness evaluated. POTENTIAL RELEVANCE: It may be possible to define a substrate stiffness 'tuned' to a horse's body enabling maximal energetic economy when running. The differences between treadmills allows more accurate comparisons between physiological studies conducted on treadmills of different stiffness, and might help to identify an ideal track stiffness to reduce locomotor injuries in equine athletes. PMID- 17402450 TI - Method for quantifying net anaerobic power in exercising horses. AB - REASON FOR PERFORMING STUDY: There is no good method for measuring net anaerobic power in exercising horses to allow accurate estimates of total metabolic power. HYPOTHESIS: The increase in VO2max when breathing hyperoxic (HO) gas should be accompanied by a stoichiometrically equal (in terms of ATP turnover, i.e. energy equivalents) decrease in plasma lactate accumulation rate (Mlactate). METHODS: Six 3-year-old Thoroughbreds were trained on an equine treadmill wearing a semi open flow mask for measurement of VO2. After 4 months the horses ran with reproducible specific VO2max (VO2max/kg bwt). The mask design allowed mixing of O2 or N2 with the inward bias flow of gas so that inspired O2 concentration of the horse could be controlled. While the horse breathed either HO (25.1% O2), normoxic (NO, 21% O2) or hypoxic (LO, 19.5% O2) gas, it ran at a speed sufficient to elicit VO2max in NO while jugular venous blood was drawn at 15 sec intervals over a period of 2 min to determine Mlactate. RESULTS: VO2max/kg bwt was not significantly different between LO and NO conditions, and LO data could not be used in the comparison. The VO2max/kg bwt increased from 2.59 +/- 0.24 (s.d.) to 2.86 +/- 0.24 mlO2 (STPD)/sec/kg in NO and HO, respectively, while Mlactate decreased from 11.5 +/- 4.2 to 9.0 +/- 3.9 mmol/min as VO2 increased. CONCLUSIONS: The ratio of delta Mlactate to delta VO2max/kg bwt suggests that Mlactate of approx 11.1 +/- 6.7 mmol/min is associated with net anaerobic power approximately equivalent to 1.0 mlO2 (STPD)/sec/kg of aerobic power (20.1 W/kg( 1)). The high variability in VO2max/kg bwt observed in data from some runs, particularly in LO, suggests that caution must be used when comparing data from the same horse during different runs. POTENTIAL RELEVANCE: This study provides a tool for estimating net anaerobic power and, more accurately, evaluating total metabolic power of horses exercising at or above their aerobic capacities. PMID- 17402451 TI - Effects of egg-bar shoes on the 3-dimensional kinematics of the distal forelimb in horses walking on a sand track. AB - REASONS FOR PERFORMING STUDY: Understanding of the biomechanical effects of egg bar shoes remains incomplete because kinematic studies are usually performed on hard tracks and with skin markers that do not measure the actual 3-dimensional (3D) movements of the 3 digital joints. OBJECTIVE: To quantify the effects of egg bar shoes on the 3D kinematics of the distal forelimb in horses walking on a sand track. METHODS: Four healthy horses were equipped with ultrasonic markers fixed surgically to the 4 distal segments of the left forelimb. The 3D movements of these segments were recorded while the horses were walking on a sand track. Rotations of the digital joints were calculated by use of a joint coordinate system. Data obtained with egg-bar shoes were compared to those obtained with standard shoes. Mean differences were expressed in a 0.95 confidence interval. RESULTS: With egg-bar shoes, the initial sinking of the heels into the ground during landing was reduced and the heels were raised by up to 5.1 degrees (3.5 6.7 degrees) compared to standard shoes at mid-stance. Concurrently, maximal flexion of the distal (DIPJ) and proximal (PIPJ) interphalangeal joints was increased by up to 3.2 degrees (2.2-4.2 degrees) and 1.8 degrees (1.1-2.5 degrees), respectively, at the beginning of the stance phase. At heel-off, extension of the DIPJ was reduced by 3.8 degrees (2.6-5.0 degrees). In extrasagittal planes of movement, egg-bar shoes prevented sinking of the medial quarter into the ground which led to a slight decrease of DIPJ medial rotation and lateromotion. CONCLUSIONS: Egg-bar shoes prevent the heels and, to a lesser extent, the medial side of the hoof from sinking into the ground on a sand track. They contribute to a decrease of DIPJ maximal extension at heel-off and to hoof stabilisation in the transversal plane. POTENTIAL RELEVANCE: Such quantitative results support the clinical indications of egg-bar shoes. PMID- 17402452 TI - Estimating peak strains associated with fast exercise in thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: The third metacarpal bones (MC3) of racing Thoroughbreds are a common site for bone damage. The midshaft dorsal cortex (DC) of MC3 thickens in response to fast exercise. It is not clear if it changes to a shape and size that limits the peak bone strains to a range associated with normal loading in other species. OBJECTIVES: To relate the proportionate size of the DC cortex in racehorses exercising at racing speed to surface strains, and test whether the DC reached a plateau that was sustained through subsequent exposures to racing speed exercise. METHODS: Standardised lateral MC3 radiographs were taken weekly for 2-5 years in 40 racehorses in race training (16-19 m/sec). DC, medulla (M), and palmar cortex (PC) thickness were measured. at 2.5 cm distal to the nutrient foramen. An index (RI) of the relative thickness of the DC was calculated for each radiograph (RI = [DC+PC]/ M multiplied by [DC/ PC]) and used to calculate strains at 12 m/sec from an equation published previously. RESULTS: Mean time to plateau in DC thickness was 501 days, mean increase in DC was 3.9 mm on the left and 3.7 mm on the right. Rate of change was 8.8 microm/day on the left, and 8.4 microm/day on the right during this time. In most horses the medulla width did not change between the first and last measurements so these bone growth rates reflect periosteal bone growth on the DC surface. No further significant change in DC or RI was found, once they had reached a plateau. Mean DC thickness at the plateau was 14.7 mm on the left and 14.9 mm on the right. Mean RI at the plateau was 3.7. This equated to peak microstrains at 12 m/sec of 2492 and suggests that strains much greater than 3000 microstrains occur at racing speed in most horses. CONCLUSIONS: Experienced racehorses are likely to be exposed to peak strains beyond the normal limit for adult mammalian bone to resist without damage and strains associated with very fast exercise may not be sufficient stimulus to induce further bone modelling in these horses. POTENTIAL RELEVANCE: Strains occur in the bones of experienced racehorses at a higher level than normal for other mammalian bones. Hence there may be other mechanisms operating in this particular bone in racehorses that protects the bone from failure when exposed to the high strains associated with fast exercise speeds. Investigation of such mechanisms may provide an insight on how to reduce the likelihood of damage to this bone during very fast exercise. PMID- 17402453 TI - Effect of head and neck position on vertical ground reaction forces and interlimb coordination in the dressage horse ridden at walk and trot on a treadmill. AB - REASONS FOR PERFORMING STUDY: Little is known in quantitative terms about the influence of different head-neck positions (HNPs) on the loading pattern of the locomotor apparatus. Therefore it is difficult to predict whether a specific riding technique is beneficial for the horse or if it may increase the risk for injury. OBJECTIVE: To improve the understanding of forelimb-hindlimb balance and its underlying temporal changes in relation to different head and neck positions. METHODS: Vertical ground reaction force and time parameters of each limb were measured in 7 high level dressage horses while being ridden at walk and trot on an instrumented treadmill in 6 predetermined HNPs: HNP1 - free, unrestrained with loose reins; HNP2 - neck raised, bridge of the nose in front of the vertical; HNP3 - neck raised, bridge of the nose behind the vertical; HNP4 - neck lowered and flexed, bridge of the nose considerably behind the vertical; HNP5 - neck extremely elevated and bridge of the nose considerably in front of the vertical; HNP6 - neck and head extended forward and downward. Positions were judged by a qualified dressage judge. HNPs were assessed by comparing the data to a velocity matched reference HNP (HNP2). Differences were tested using paired t test or Wilcoxon signed rank test (P<0.05). RESULTS: At the walk, stride duration and overreach distance increased in HNP1, but decreased in HNP3 and HNP5. Stride impulse was shifted to the forehand in HNP1 and HNP6, but shifted to the hindquarters in HNP5. At the trot, stride duration increased in HNP4 and HNP5. Overreach distance was shorter in HNP4. Stride impulse shifted to the hindquarters in HNP5. In HNP1 peak forces decreased in the forelimbs; in HNP5 peak forces increased in fore- and hindlimbs. CONCLUSIONS: HNP5 had the biggest impact on limb timing and load distribution and behaved inversely to HNP1 and HNP6. Shortening of forelimb stance duration in HNP5 increased peak forces although the percentage of stride impulse carried by the forelimbs decreased. POTENTIAL RELEVANCE: An extremely high HNP affects functionality much more than an extremely low neck. PMID- 17402454 TI - Functional anatomy of the caudal thoracolumbar and lumbosacral spine in the horse. AB - REASONS FOR PERFORMING STUDY: Research in spinal biomechanics and functional anatomy has advanced back pain research in man. Yet, despite the performance limiting nature of back pain in horses, there are few data for the equine spine. OBJECTIVES: To describe aspects of functional anatomy of the equine thoracolumbar and lumbosacral (LS) spine and potential effects on performance. METHODS: The first study investigated variations in LS vertebral formula by post mortem examination of 120 horses. Midline vertebral transection was carried out on 65 Thoroughbred (TB), 24 Standardbred (SB) and 31 other breeds. The second study investigated morphology and biomechanics of the deep stabilising epaxial muscles of 13 horses using MRI (n = 3), anatomical dissection (n = 11) and biomechanical analysis (n = 6). The spinous process angular orientation relative to the vertebral body, was analysed at vertebrae T13, T18, L3, L5, L6 and S1. RESULTS: LS variations were found in 33.3% of the total group, 40.0% TB and 45.2% others, but 0% SB. Sacralisation of lumbar vertebra (L) 6 with LS motion between L5 and L6 occurred in 32.3% TB and 29.0% others. Five segmental multifidus fascicles were identified originating from spinous processes and vertebral laminae running craniocaudally onto the mammillary processes and lateral border of the sacrum, crossing between 1-5 intervertebral discs. Sacrocaudalis dorsalis (SCD) lateralis muscle was an extension of multifidus from L4, L5 and L6 depending on the vertebral formula whereas SCD medialis mm originated from S3. Both inserted on caudal vertebrae. Based on the location and direction of fibres, the principal action of the deep epaxial muscles was dorsoventral sagittal rotation. This action was dependent on vertebral spinous process/body orientation. We hypothesise that equine multifidus and SCD lateralis muscles act as caudal sagittal rotators of their vertebra of origin, as is the case in man, allowing dynamic stabilisation during dorsoventral motion. CONCLUSION: Equine multifidus anatomy and function are comparable to that of man. The high prevalence of anatomical variations in the LS spine may affect maximal dorsoventral motion, the stability of the LS joint and, therefore, have consequences for athletic performance. Further studies of these structures are warranted in appropriately selected poorly performing horses. PMID- 17402455 TI - A pattern recognition approach for the quantification of horse and rider interactions. AB - REASONS FOR PERFORMING STUDY: Interactions of various systems were investigated in several studies of dynamic systems, but the interactions between horse and rider have not yet been documented. These interactions include the rider's ability to control the horse, adapt to the horse and maintain both participants' body position. An optimum interaction is also adapted to the individual nature of the horse. OBJECTIVE: To identify rider-horse interactions by means of artificial neural nets analysing the time-continuous pattern. METHODS: Fourteen horses were measured trotting on hand, and ridden at working trot with a professional and a recreational rider using a 3D high speed video system (120 Hz)1. Angles were calculated after low pass filtering (5-20 Hz). Horse movements were described by 2D angles, angular velocities, and angular accelerations of variables of the right body side: hind and front fetlock, head, back and the summation angle of carpus, elbow, and shoulder, the summation angle of hock, stifle, and hip. Distances between the trajectories of the feature vectors in an N = 11 x 11 Kohonen map were determined and analysed by means of a cluster analysis. RESULTS: Depending on the variables included, both rider specific as well as horse specific movement patterns could be identified. The time courses of the head angle indicate a movement pattern mainly dominated by the rider, whereas the time courses of variables of the hind fetlock and hock in most cases did not show differences between the conditions with, and without, rider. The skill of the professional rider could be documented with a higher adaptation to the horse's movement pattern. CONCLUSION AND POTENTIAL RELEVANCE: The presented time course oriented approach provides a sensitive tool in order to quantify the interaction of rider and horse. PMID- 17402456 TI - Evaluation of the force acting on the back of the horse with an English saddle and a side saddle at walk, trot and canter. AB - REASONS FOR PERFORMING STUDY: Force transmission under an English saddle (ES) at walk, trot and canter is commonly evaluated, but the influence of a side saddle (SS) on the equine back has not been documented. HYPOTHESIS: Force transmission under a SS, with its asymmetric construction, is different from an ES in walk, trot and canter, expressed in maximum overall force (MOF), force in the quarters of the saddle mat, and centre of pressure (COP). The biomechanics of the equine back are different under a SS compared to ES. METHODS: Thirteen horses without clinical signs of back pain ridden in an indoor riding school with both saddles were measured using an electronic saddle sensor pad. Synchronous kinematic measurements were carried out with tracing markers placed along the back in front of (withers, W) and behind the saddle (4th lumbar vertebra, L4). At least 6 motion cycles at walk, trot and canter with both saddles (ES, SS) were measured. Out of the pressure distribution the maximum overall force (MOF) and the location of the centre of pressure (COP) were calculated. RESULTS: Under the SS the centre of pressure was located to the right of the median and slightly caudal compared to the COP under the ES in all gaits. The MOF was significantly different (P<0.01) between saddles. At walk, L4 showed significantly larger (P<0.01) vertical excursions under the ES. Under the SS relative horizontal movement of W was significantly reduced (P<0.01) at trot, and at canter the transversal movement was significantly reduced (P<0.01) . In both trot and canter, no significant differences in the movement of L4 were documented. CONCLUSIONS AND POTENTIAL RELEVANCE: The results demonstrate that the load under a SS creates asymmetric force transmission under the saddle, and also influences back movement. To change the load distribution on the back of horses with potential back pain and as a training variation, a combination of both riding styles is suitable. PMID- 17402457 TI - Association of type of sport and performance level with anatomical site of orthopaedic injury diagnosis. AB - REASON FOR PERFORMING STUDY: Although anecdotal reports of increased orthopaedic injury risk in equine sports exist, there is little scientific evidence to support this. OBJECTIVES: To test whether horses undertaking a single competitive sport have increased risk of specific injuries compared to those used for general purpose riding (GP); and whether injury type varies with sport category and performance level. METHODS: Data from 1069 records of horses undergoing orthopaedic evaluation (1998-2003) and meeting inclusion criteria were reviewed. Sport category (GP, showjumping, dressage, eventing, racing), level (nonelite or elite) and diagnosis were recorded. Effects of sport category and level on probability of a specific diagnosis were assessed using chi-squared tests. Logistic regression was used to determine which competitive sports and levels increased risk of injury compared with GP. RESULTS: Overall there was a significant effect of sport category and level on diagnosis (P<0.0001). There was significant difference between anatomical site injured and sport category (P<0.0001); a high risk of forelimb superficial digital flexor tendon injury in elite eventing (P<0.0001) and elite showjumping (P = 0.02); distal deep digital flexor tendon (DDFT) injury in elite showjumping (P = 0.002); and hindlimb suspensory ligament injury in elite (P<0.0001) and nonelite (P = 0.001) dressage. There was a low risk of tarsal injury in elite eventing (P = 0.01) and proximal DDFT injury in dressage (P = 0.01). CONCLUSIONS: Horses competing in different sports are predisposed to specific injuries; particular sports may increase the risk of injury at certain anatomical sites; and the type and site of injury may reflect the type and level of performance. POTENTIAL RELEVANCE: These findings could guide clinicians in the diagnosis of sport related injuries. PMID- 17402458 TI - Abnormal radiographic findings in 865 French standardbred trotters and their relationship to racing performance. AB - REASON FOR PERFORMING STUDY: Developmental orthopaedic lesions are commonly found in French Standardbred horses. One of the main questions asked by trainers, owners and veterinarians is what impact these lesions have on the racing career and racing performances of horses. OBJECTIVES: To study the prevalence and distribution of developmental orthopaedic lesions in young French Standardbred trotters and to relate them to racing performance. METHODS: Feet, fetlock, tarsus and stifle regions were radiographed in 865 two-year-old French Standardbred trotters. Abnormal radiographic findings (ARF) were evaluated for 12 anatomical sites identified in these areas, and a severity index given. Performance criteria were: success in qualification for racing, maximal and mean index of trot (ITR), an annual index calculated on the basis of the logarithm of earnings per starts, total earnings at 5 years, placed races compared to starts and longevity of the racing career. Analysis of variance were calculated to study the relationships between racing performance and the number of ARF or the severity index. RESULTS: A total of 363 horses (42.0%) showed ARF. Prevalence of ARF was 18.3% in the plantar aspect of the hind fetlock and 10.6% in the proximal tarsus. Among the total population, 833 horses were considered for performance evaluation, 478 of them were qualified for racing. The number of ARF significantly affected racing longevity. However, the number of ARF did not affect performance categories according to maximal ITR. Concerning distribution of ARF, the number of plantar lesions in the fetlock significantly affected mean ITR. The index of severity did not provide more information for prognosis than the number of ARF. CONCLUSION: Longevity is the only criteria affected by ARF. When evaluating different sites, only the plantar fetlock region showed a significant relationship with mean ITR. POTENTIAL RELEVANCE: Number of ARF and radiographic score (RS) affect mean ITR and longevity but do not affect maximal ITR. A horse with a good racing ability will be a good performer but might have a racing career shortened because of orthopaedic problems in relation to developmental orthopaedic lesions. PMID- 17402459 TI - Gait pattern of the ataxic horse compared to sedated and nonsedated horses. AB - REASONS FOR PERFORMING STUDY: Equine ataxia is routinely evaluated subjectively by clinicians; however, objective measurements of the movement and coordination of ataxic horses have not been reported. OBJECTIVES: To document the movement pattern of ataxic horses and compare the results to the movement of neurologically sound horses with, and without, sedation. METHODS: Seventeen ataxic horses were evaluated walking and trotting on a treadmill using a 3D high speed video system. From the horizontal movement of hoof markers the autocorrelation function (ACF) of the left forelimb and the cross correlation function (CCF) between the left forelimb and other 3 limbs (CCF foreleft/hindright [flhr], foreleft/ hindleft [flhl], foreleft/foreright [flfr]) was calculated. This resulted in a value of motion cycle consistency. The results were compared to data of 17 neurologically normal horses with, and without, sedation with detomidine. For statistical analysis the t test for independent samples was used. RESULTS: Comparing normal (NO) and ataxic (AT) horses at the walk, highly significant differences for the ACF and for all the CCFs were documented. At the trot, ACF and CCFs flhr and flfr were significantly different in the NO and AT groups. Comparing sedated and ataxic horses, only the CCF flfr at walk and at trot were significantly different. CONCLUSION AND POTENTIAL RELEVANCE: Ataxia is best documented and examined at the walk. At trot pendulum effect may make coordination easier for horses. The results of this study serve as basic data for evaluating questionably ataxic horses. PMID- 17402460 TI - Raising heels of hind hooves changes the equine coffin, fetlock and hock joint angle: a kinematic evaluation on the treadmill at walk and trot. AB - REASONS FOR PERFORMING STUDY: Raised heels are commonly recommended for various equine orthopaedic conditions. However, the simultaneous effect of raised heels on the different joint angles of the equine hindlimb throughout the motion cycle has not been previously evaluated. OBJECTIVE: To document the simultaneous effect of raised heels on the joint angles of the equine hindlimb coffin, fetlock and hock joints. METHODS: Eight sound, adult, Warmblood horses were evaluated barefoot and with a heel wedge of 8 or 16 degrees, walking and trotting on a horizontal treadmill. Markers placed on the dorsal and cranial aspect of the hindlimb were traced using a 3D high speed video system and joint angles calculated. RESULTS: The effects of raising the hindlimb heels by 8 or 16 degrees on the angles of the hindlimb during the stance phase are a reduction of the plantar combined coffin joint and pastern joint angle, a reduction of maximum extension in the fetlock joint, and an increase in maximum hock flexion. The relation between angles did not change significantly during the course of the stance phase in the three measurement situations, with only small differences in time of occurrence of each joint angle maxima and minima. CONCLUSIONS: Raising the heels of hind hooves increases flexion of the coffin and hock joints during the stance phases of walk and trot, and a doubling of the angle of the raised heels also doubles the effect on the joint angles investigated. Raised heels reduce the maximum extension of the fetlock joint during the the stance phase at walk and trot. POTENTIAL RELEVANCE: This study provides evidence for the therapeutic use of raised shoes with heels in horses with pain on maximum hock extension, e.g. spavin. PMID- 17402461 TI - The effect of gallop training on hoof angle in thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: The economic impact of soundness problems in racehorses is very high and low hoof angle at the toe has been associated with a lack of soundness. However, it is not clear what environmental and management factors might contribute to a low hoof angle. OBJECTIVES: To investigate the hypothesis that the hooves of racehorses become flatter when in gallop training, as well as to determine factors contributing to this trend. METHODS: Weekly hoof measurements were taken with a hoof gauge from 45 Thoroughbred racehorses; 4 Thoroughbred show horses kept in consistent conditions and shod by the same farrier as some of the racehorses; and 6 unshod free-ranging horses. A further 15 horses were measured twice in one day to determine the repeatability of the method. RESULTS: Repeatability coefficients were 0.31 degrees for the left hoof and 0.37 degrees for the right. Racehorses in training showed a significant decrease in hoof angle over time while free ranging horses and show horses did not. Free-ranging horses had a significantly lower angle in winter (wet) compared with summer (dry) in both left (P = 0.040) and right (P = 0.017). Show horses had no significant change in hoof angle. Racehorses that had a period of rest during the experiment (n = 11) showed a decrease in hoof angle during training and an increase over their rest period for both hooves (P = 0.005 for the left hoof, P = 0.0009 for the right). CONCLUSIONS: Training for fast exercise in Thoroughbred racehorses is associated with a reduction in hoof angle and wet pasture conditions may also be associated with a reduced hoof angle in free-ranging horses. POTENTIAL RELEVANCE: Gallop exercise has a potentially large effect on hoof angle and therefore, a change in angle should be expected to occur in racehorses starting fast exercise work. Hence management of horses with abnormally low hoof angles may require an adaptation to their training regime in order to minimise this effect. PMID- 17402462 TI - Effect of uphill exercise on equine superficial digital flexor tendon forces at trot and canter. AB - REASONS FOR PERFORMING STUDY: One cause of overstrain injury to the superficial digital flexor tendon (SDFT) in horses is the force loaded on the SDFT during repeated running. Therefore, decreasing this force may reduce SDFT injury. It has been reported that strain on the SDFT decreases with a toe-wedge shoe. Uphill courses are used for training of racehorses, and the angle of hoof-sole to the horizon during uphill running is similar to that of the toe-wedge shoe. OBJECTIVES: To determine the effects of uphill exercise on the force on the SDFT during trotting and cantering. METHODS: Arthroscopically implantable force probes (AIFP) were implanted into the SDFT of the left or right forelimb of 7 Thoroughbred horses and AIFP output recorded during trotting and cantering on a treadmill inclined at slopes of 0, 3 or 8%, and then 0% again. Superficial digital flexor tendon force was calculated as a relative value, with the amplitude of AIFP output voltage at initial 0% slope equal to 100. RESULTS: Out of 14 sets of experiments, AIFP data were analysed successfully in 9 at the trot, in 3 at the canter in the trailing forelimb on a slope of 3 and 8%, and in 2 at the canter in the leading forelimb on a slope of 3%. Increasing the incline from 0-8% tended to decrease peak force in the SDFT at the trot, and in the trailing forelimb at the canter. However, force in the SDFT was unchanged in the leading forelimb at the canter on the 3% incline. CONCLUSIONS: The force in the SDFT trotting or cantering uphill is unchanged or lower than that loaded at the same speed on a flat surface. Because at similar speeds the workload for uphill exercise is greater than on the flat, uphill running increases exercise intensity without increasing force in the SDFT. POTENTIAL RELEVANCE: Uphill exercise may reduce the risk of SDFT injury as both running speed and SDFT force are decreased on an incline as compared to the flat, even when exercise intensity is the same. Further study is needed to confirm these findings at canter in a larger population of horses. PMID- 17402463 TI - Determination of mechanical loading components of the equine metacarpus from measurements of strain during walking. AB - REASONS FOR PERFORMING STUDY: The mechanical environment of the distal limb is thought to be involved in the pathogenesis of many injuries, but has not yet been thoroughly described. OBJECTIVES: To determine the forces and moments experienced by the metacarpus in vivo during walking and also to assess the effect of some simplifying assumptions used in analysis. METHODS: Strains from 8 gauges adhered to the left metacarpus of one horse were recorded in vivo during walking. Two different models - one based upon the mechanical theory of beams and shafts and, the other, based upon a finite element analysis (FEA) - were used to determine the external loads applied at the ends of the bone. RESULTS: Five orthogonal force and moment components were resolved by the analysis. In addition, 2 orthogonal bending moments were calculated near mid-shaft. Axial force was found to be the major loading component and displayed a bi-modal pattern during the stance phase of the stride. The shaft model of the bone showed good agreement with the FEA model, despite making many simplifying assumptions. CONCLUSIONS: A 3 dimensional loading scenario was observed in the metacarpus, with axial force being the major component. POTENTIAL RELEVANCE: These results provide an opportunity to validate mathematical (computer) models of the limb. The data may also assist in the formulation of hypotheses regarding the pathogenesis of injuries to the distal limb. PMID- 17402464 TI - The effect of head and neck position on the thoracolumbar kinematics in the unridden horse. AB - REASONS FOR PERFORMING STUDY: In many equestrian activities a specific position of head and/or neck is required that is dissimilar to the natural position. There is controversy about the effects of these positions on locomotion pattern, but few quantitative data are available. OBJECTIVES: To quantify the effects of 5 different head and neck positions on thoracolumbar kinematics of the horse. METHODS: Kinematics of 7 high level dressage horses were measured walking and trotting on an instrumented treadmill with the head and neck in the following positions: HNP2 = neck raised, bridge of the nose in front of the vertical; HNP3 = as HNP2 with bridge of the nose behind the vertical; HNP4 = head and neck lowered, nose behind the vertical; HNP5 = head and neck in extreme high position; HNP6 = head and neck forward and downward. HNP1 was a speed-matched control (head and neck unrestrained). RESULTS: The head and neck positions affected only the flexion-extension motion. The positions in which the neck was extended (HNP2, 3, 5) increased extension in the anterior thoracic region, but increased flexion in the posterior thoracic and lumbar region. For HNP4 the pattern was the opposite. Positions 2, 3 and 5 reduced the flexion-extension range of motion (ROM) while HNP4 increased it. HNP5 was the only position that negatively affected intravertebral pattern symmetry and reduced hindlimb protraction. The stride length was significantly reduced at walk in positions 2, 3, 4 and 5. CONCLUSIONS: There is a significant influence of head/neck position on back kinematics. Elevated head and neck induce extension in the thoracic region and flexion in the lumbar region; besides reducing the sagittal range of motion. Lowered head and neck produces the opposite. A very high position of the head and neck seems to disturb normal kinematics. POTENTIAL RELEVANCE: This study provides quantitative data on the effect of head/neck positions on thoracolumbar motion and may help in discussions on the ethical acceptability of some training methods. PMID- 17402465 TI - Effects of draw reins on limb kinematics. AB - REASONS FOR PERFORMING STUDY: No data exist on the GRF-kinematics relation due to changes caused by equestrian interventions. HYPOTHESIS: Through the judicious use of draw reins the rider can influence the kinematics of the horse to meet stated goals of dressage training. Relating the results to previously published kinetic data of the same experiment implies a possible relationship between kinetics and kinematics. METHODS: The kinematics of 8 sound Swedish Warmblood horses were measured whilst the horses were being ridden with and without draw reins. Three conditions were evaluated: 1) draw reins only (DR), 2) combination of draw reins and normal reins (NR+DR) and 3) normal reins only (NR). RESULTS: Head and neck angles were significantly decreased by the draw rein but 4-5 times more so for DR when with NR+DR. The forelimb position at hoof lift-off was significantly more caudal with DR. In the hind limb the hip joint extended more quickly and the hock joint flexed more with NR+DR than with NR. Compared to DR the hip joint angular pattern was not significantly different, but the pelvis was more horizontal. CONCLUSION: Riding with a draw rein can have significant influence on the kinematics of the horse. Some of the observed changes can be coupled to changes in kinetics. The hock joint angle seems to be a fairly reliable indicator of load on the hind limb and the angle of femur appears important for hind limb propulsion, when considered in conjunction with the orientation of the pelvis. POTENTIAL RELEVANCE: These findings are important for riders and trainers, as kinematic changes are what trainers observe. It is thereby important to ascertain which kinematic changes are consistently coupled to changes in kinetics in order for trainers to be able to judge correctly the success of intended goals. Further studies are warranted to validate and confirm suggested relationships between kinetics and kinematics. PMID- 17402466 TI - Movement between the equine ilium and sacrum: in vivo and in vitro studies. AB - REASONS FOR PERFORMING STUDY: Sacroiliac disease (SID) is an important cause of loss of performance in horses, yet little is known about the biomechanics of movement of the sacroiliac joint (SIJ). OBJECTIVES: (a) To document the presence of equine SIJ motion by measuring the change in cross-sectional area (CSA) of the dorsal portion of the dorsal sacroiliac ligament (DSIL) during application of manual forces to the pelvis, and to ascertain if this varied between horses with SID and normal horses; (b) To determine the amount and direction of motion available at the equine SIJ using 3-D orientation sensors, and how motion is limited by the DSIL and sacrotuberous ligament. METHODS: Study 1: CSA obtained ultrasonographically was compared before and during manual force application to the ilium, on a group of 10 horses, 5 with SID and 5 clinically normal. Study 2: direction and degrees of motion between sacrum and ilium were measured in 8 cadaveric SIJs. 3D orientation sensors were mounted to the fixated sacrum and the moveable ilium, and relative motion recorded between the 2 bones when manual forces were applied to the ilium. RESULTS: Study 1 showed a significant decrease in the CSA of the equine DSIL during application of manual forces to both tuber coxae (TC) (P<0.001) and tuber sacrale (TS) (P<0.001) when compared to at rest. Study 2 described range of motion to be greatest in the transverse or coronal plane, when lateral and oblique forces were applied to the pelvis, and recorded significant increases (P<0.05) in range of motion in the sagittal plane following resection of both the DSIL and sacrotuberous ligament. CONCLUSION AND POTENTIAL RELEVANCE: The change in DSIL CSA with application of manual force may be due to a lengthening of the ligament, reflecting equine SIJ motion in vivo. The motion recorded between ilium and sacrum in vitro suggests that greatest sacroiliac motion may occur in the transverse plane. Increases in range of motion following resection of ligaments give some indication of the role of each ligament. PMID- 17402467 TI - Head and body centre of mass movement in horses trotting on a circular path. AB - REASONS FOR PERFORMING STUDY: Horses are often worked along a circular trajectory but the mechanics of turning have not been reported. OBJECTIVE: To develop a methodology for kinematic analysis of horses during circling and to report preliminary findings describing movements of the total body centre of mass (COMbody). METHODS: Seventy-five skin markers, distributed over the head, neck, trunk and limbs of 5 sound horses, were tracked in a global coordinate system at 120 Hz as the horses trotted around a small circle. Based on segmental inertial data, 3D positions of the segmental centres of mass and COMbody were calculated. Movements of COMbody were measured relative to the cylindrical vertical plane, the radius of which was defined by a marker overlying the sixteenth thoracic vertebra. RESULTS: During trotting, COMbody was high at the start and end of the diagonal stance phases and low in midstance. All horses leaned to the inside of the circle, with a tilt angle of mean +/- s.d. 14.8 +/- 2.8 degrees during the entire stride. In the transverse direction, COMbody swung from side to side reaching its most inward position around the time of contact of the inside forelimb and its most outward position around the time of contact of the outside forelimb. The centre of mass of the head and neck segments (COMhead/neck) had a symmetrical, double sinusoidal pattern in the vertical direction. All horses had the same motion pattern for COMhead/neck in the transverse direction but the mean position relative to the circle line varied between horses. CONCLUSIONS: Horses lean to the inside when turning around a small circle. COMhead/neck and COMbody showed 2 vertical oscillations in each stride, whereas there was a single mediolateral excursion that was not symmetrical to the inside and outside of the circle line. POTENTIAL RELEVANCE: This study provides a first step toward understanding the mechanics of locomotion during turning, which has implications in the aetiology of injuries, the response to lungeing as a diagnostic technique and the incorporation of circles into rehabilitation exercises. PMID- 17402468 TI - Harnessed vs. mounted standardbreds on the track: changes in gait and physiological variables. AB - REASONS FOR PERFORMING STUDY: It has become apparent through analysis of elite races over the past 20 years that mounted races are always slower than harnessed races when performed by the same horses over the same distances on the same tracks. OBJECTIVES: To investigate and compare physiological and gait variables in mounted and harnessed activities in trotters. METHODS: Ten trotters were taken at random in 2 standardised field exercise tests one week apart: a harnessed and a mounted test with standardised weight jockeys and drivers. Speed, heart rate (HR), respiratory frequency (RF), blood lactate concentration (La), stride characteristics (length, frequency [SF], symmetry, regularity, dorsoventral displacement of the sternum and vertical, longitudinal and lateral activities measuring the amount of deceleration and acceleration along the 3 axis) were measured. RESULTS: Paired t tests and an analysis of variance were calculated between the 2 conditions and revealed that V4 (speed for a La of 4 mmol/I), V200 (speed for a HR of 200 beats/min), stride length, trot symmetry, lateral activity were lower in the mounted condition. In contrast, RF, SF, dorsoventral displacement, vertical and longitudinal activities were significantly higher in mounted horses compared to harnessed horses. CONCLUSION: The significant physiological and biomechanical differences between pulling whilst harnessed and being ridden reflect an increase in the energy requirement of locomotion in the mounted condition compared to the harnessed condition. Trotting mounted at maximal speed is more demanding for the horse than harnessed as reflected by the lower V4 and V200 and some gait variables change in relation to this phenomenon. POTENTIAL RELEVANCE: This study demonstrates that trotting mounted at sub-maximum and maximum speed is more demanding for the horse when ridden and that it modifies some gait variables, either as a result or even as a possible cause. PMID- 17402469 TI - Correlation between routine radiographic findings and early racing career in French trotters. AB - REASONS FOR PERFORMING STUDY: The relationship between the presence of radiological abnormalities and subsequent racing performance is controversial. However, as training is expensive and time consuming, it would save time and money to identify subjects with osteo-articular lesions not compatible with a normal racing career on the basis of routine radiographic screenings at yearling age. OBJECTIVES: To evaluate the impact of osteo-articular lesions on racing ability in French Trotters and identify radiographic changes associated with failure in 'qualification', in order to provide objective criteria for selection of horses based on their osteo-articular status. HYPOTHESIS: The influence of radiographic findings (RF) on racing ability depends on their nature, location, clinical relevance and number. METHODS: The limbs of 202 French Trotters were radiographed just before they started training. All the RF were graded according to a standardised protocol depending on their severity. The success in 'qualification' (first race in career of French Trotters) was the criteria used to assess racing ability. Breeders and trainers were questioned about the causes for horses not racing. RESULTS: Overall 113 (55.9%) horses qualified. Osteoarticular lesions were directly responsible for nonqualification in 31% of the horses. Subjects with more than one abnormal RF, with abnormal RF on the fore , hind-fetlock or proximal tarsus were less likely to qualify. Dorsal modelling in the front fetlock and osteochondrosis of the lateral trochlear ridge of the femur also significantly reduced the qualification rate. CONCLUSIONS: Most RF are compatible with beginning a racing career, but severe RF or multiple abnormal RF significantly compromise future racing career. POTENTIAL RELEVANCE: This study supports the use of routine radiographic programmes for detection of osteoarticular lesions in yearlings. A synthetic radiographic score, based on both the severity and the number of lesions, could be useful for breeders and trainers as complementary information to select their horses. PMID- 17402470 TI - Nitric oxide synthase inhibition in thoroughbred horses augments O2 extraction at rest and submaximal exercise, but not during short-term maximal exercise. AB - REASON FOR PERFORMING STUDY: Work is required to establish the role of endogenous nitric oxide (NO) in metabolism of resting and exercising horses. OBJECTIVES: To examine the effects of NO synthase inhibition on O2 extraction and anaerobic metabolism at rest, and during submaximal and maximal exertion. METHODS: Placebo and NO synthase inhibition (with Nomega-nitro-L-arginine methyl ester [L-NAME] administered at 20 mg/kg bwt i.v.) studies were performed in random order, 7 days apart on 7 healthy, exercise-trained Thoroughbred horses at rest and during incremental exercise leading to 120 sec of maximal exertion at 14 m/sec on a 3.5% uphill grade. RESULTS: At rest, NO synthase inhibition significantly augmented the arterial to mixed-venous blood O2 content gradient and O2 extraction as mixed venous blood O2 tension and saturation decreased significantly. While NO synthase inhibition did not affect arterial blood-gas tensions in exercising horses, the exercise-induced increment in haemoglobin concentration and arterial O2 content was attenuated. In the L-NAME study, during submaximal exercise, mixed-venous blood O2 tension and haemoglobin-O2 saturation decreased to a greater extent causing O2 extraction to increase significantly. During maximal exertion, arterial hypoxaemia, desaturation of haemoglobin and hypercapnia of a similar magnitude developed in both treatments. Also, the changes in mixed-venous blood O2 tension and haemoglobin-O2 saturation, arterial to mixed-venous blood O2 content gradient, O2 extraction and markers of anaerobic metabolism (lactate and ammonia production, and metabolic acidosis) were not different from those in the placebo study. CONCLUSION: Endogenous NO production augments O2 extraction at rest and during submaximal exertion, but not the during short-term maximal exercise. Also, NO synthase inhibition does not affect anaerobic metabolism at rest or during exertion. POTENTIAL RELEVANCE: It is unlikely that endogenous NO release modifies aerobic or anaerobic metabolism in horses performing short-term maximal exertion. PMID- 17402471 TI - Evaluation of race distance, track surface and season of the year on exercise induced pulmonary haemorrhage in flat racing thoroughbreds in Brazil. AB - REASONS FOR PERFORMING STUDY: To our knowledge no studies evaluating risk factors to EIPH have been undertaken with such a population of horses in Brazil, which is one of the few countries in the racing community to accept the use of frusemide (FUR) as a prerace medication for bleeders. We also compared national results with those already in the literature. OBJECTIVES: To verify the influence of variables such as distance of the race, racetrack surface, going and season of the year in the recurrence and severity of EIPH and its effect on the finishing position of flat racing Thoroughbreds in Brazil. METHODS: 2118 post race respiratory endoscopies were recorded, in a total of 1003 individual horses. All horses in the study were certified bleeders (deemed positive by registered veterinarians following one or more endoscopies) and were administered FUR prior to each race. Data was analysed using both ordinary logistic regression and multiple logistic regression. RESULTS: The results suggest that race distance, season of the year and racetrack surface affect the recurrence of EIPH. Also, despite the use of FUR, the majority (62%) of horses continued to display some degree of haemorrhage. CONCLUSIONS: There was a tendency for horses that remained positive despite the use of FUR to finish unplaced in a race. Horses which had a significant reduction in bleeding and became HPIE negative after the use of FUR, tended to finish in top positions. It was not possible to determine a pattern of severity of EIPH in horses with more than one endoscopy. Further studies are required to improve our understanding of this syndrome and its correlations to performance in the racing Thoroughbred, together with the role of FUR and its possible enhancing performance effects and efficacy in consistently diminishing EIPH. POTENTIAL RELEVANCE: This study enabled a better understanding of the use of FUR in Brazilian Thoroughbred racehorses and showed there was a tendency towards improvement in performance after administrion of FUR in bleeders. The fact that a large number of horses remained positive for EIPH despite the use of FUR should encourage the scientific community to search for better ways of reducing, or preventing, this syndrome. PMID- 17402472 TI - The effect of inspired gas density on pulmonary artery transmural pressure and exercise induced pulmonary haemorrhage. AB - REASONS FOR PERFORMING STUDY: Pulmonary capillary stress failure, largely as a result of high pulmonary vascular pressures, has been implicated in the aetiology of EIPH. However, the role of the respiratory system in determining the magnitude of EIPH has received little attention. HYPOTHESIS: Horses breathing a gas of greater density than air will exhibit greater transmural pulmonary arterial pressures (TPAP) and more severe EIPH, and horses breathing a gas of lower density than air will exhibit lower TPAP and less severe EIPH, both compared with horses breathing air. METHODS: Following a warm-up, 8 Thoroughbred horses were exercised for 1 min at 10, 11 and 12 m/sec (5 degrees incline) breathing air or 21% oxygen/79% helium or 21% oxygen/79% argon in a randomised order. Heart rate, respiratory rate, pulmonary arterial pressure and oesophageal pressure were measured during exercise. Bronchoalveolar lavage fluid (BALF) was collected from the dorsocaudal regions of the left and right lungs 40 min post exercise and red blood cell (RBC) counts were performed. RESULTS: The exercise tests induced mild EIPH. Maximum changes in oesophageal pressure were lower on helium-oxygen compared to argon-oxygen (P<0.001). TPAP and median RBC counts did not differ between gas mixtures. BALF RBC counts from the left lung correlated with counts from the right lung (P<0.0001). However BALF RBC counts from the left lung were higher than those from the right lung (P = 0.004). CONCLUSION: As alterations in pulmonary arterial and oesophageal pressure caused by changes in inspired gas density were of similar magnitude, TPAP remained unchanged and there was no significant effect on EIPH severity. POTENTIAL RELEVANCE: Manipulations that decrease swings in intrapleural pressure may only decrease the degree of EIPH in horses severely affected by the condition. PMID- 17402473 TI - Pre-exercise hypervolaemia is not detrimental to arterial oxygenation of horses performing a prolonged exercise protocol simulating the second day of a 3-day equestrian event. AB - REASONS FOR PERFORMING STUDY: Hyperhydration, prior to prolonged moderate intensity exercise simulating the 2nd day of a 3-day equestrian event (E3DEC), may induce arterial hypoxaemia detrimental to performance. OBJECTIVES: Because moderate-intensity exercise does not induce arterial hypoxaemia in healthy horses, the effects of pre-exercise hypervolaemia on arterial oxygenation were examined during a prolonged exercise protocol. METHODS: Blood-gas studies were carried out on 7 healthy, exercise-trained Thoroughbred horses in control and hyperhydration experiments. The study conformed to a randomised crossover design. The sequence of treatments was randomised for each horse and 7 days were allowed between studies. Hyperhydration was induced by administering 0.425 g/kg bwt NaCl via nasogastric tube followed by free access to water. The exercise protocol was carried out on a treadmill set at a 3% uphill grade and consisted of walking at 2 m/sec for 2 min, trotting for 10 min at 3.7 m/sec, galloping for 2 min at 14 m/sec (which elicited maximal heart rate), trotting for 20 min at 3.7 m/sec, walking for 10 min at 1.8 m/sec, cantering for 8 min at 9.2 m/sec, trotting for 1 min at 5 m/sec and walking for 5 min at 2 m/sec. RESULTS: NaCl administration induced a significant mean +/- s.e. 15.5 +/- 1.1% increase in plasma volume as indicated by a significant reduction in plasma protein concentration. In either treatment, whereas arterial hypoxaemia was not observed during periods of submaximal exercise, short-term maximal exertion caused significant arterial hypoxaemia, desaturation of haemoglobin, hypercapnoea, and acidosis in both treatments. However, the magnitude of exercise-induced arterial hypoxaemia, desaturation of haemoglobin, hypercapnoea, and acidosis in both treatments remained similar, and statistically significant differences between treatments could not be demonstrated. CONCLUSIONS: It was concluded that significant pre exercise expansion of plasma volume by this method does not adversely affect the arterial oxygenation of horses performing a prolonged exercise protocol simulating the 2nd day of an E3DEC. PMID- 17402474 TI - Exercise-induced pulmonary haemorrhage during submaximal exercise. AB - REASONS FOR PERFORMING STUDY: Maximally exercising horses achieve mean pulmonary artery pressures (Ppa(mean)) that exceed the minimum threshold (75 mmHg) estimated for pulmonary capillary rupture and exercise-induced pulmonary haemorrhage (EIPH). EIPH is not expected to occur during moderate submaximal exercise (i.e. 40-60% VO2max) since Ppa(mean) remains well below this threshold. HYPOTHESIS: Prolonged submaximal exercise (trotting) would precipitate locomotory respiratory uncoupling and cause EIPH. This would be present as a result of the most negative intrapleural pressures (as estimated by the minimum oesophageal pressure; Poes(min)) occurring simultaneously with the most positive Ppa (Ppa(peak)) to produce estimated maximal pulmonary artery transmural pressures (PATMPmax) that surpass the EIPH threshold. METHODS: Five Thoroughbred horses trotted to fatigue (approximately 25 min) at 5 m/sec on a 10% incline. Ventilation (V(E)), Poes, and Ppa were measured at 5 min intervals, and bronchoalveolar lavage (BAL) red blood cells (RBCs) were quantified 45 min post exercise. RESULTS: BAL revealed an increased EIPH (rest: 2.0 +/- 1 x 10(5), exercise: 17 +/- 10 x 10(5) RBCs/ml BALF; P<0.05), despite the highest Ppamean reaching only mean +/- s.e. 55 +/- 3 mmHg, while V(E), tidal volume and Poes(min) approached 70-80% of the values achieved at maximal running speeds (10% incline: 12-13 m/sec) by these same horses. The resulting PATMPmax was well above the level considered causative of EIPH. CONCLUSIONS: The finding of significant EIPH during submaximal exercise broadens the spectrum of performance horses susceptible to EIPH and supports studies that suggest that extravascular factors are of primary importance in the aetiology of EIPH. POTENTIAL RELEVANCE: Consideration of strategies such as the equine nasal strip for reducing negative extravascular pressures is warranted even for exercise at moderate intensities. PMID- 17402475 TI - Effects of chronic acetazolamide administration on fluid flux from the pulmonary vasculature at rest and during exercise in horses. AB - REASONS FOR PERFORMING STUDY: Horses develop high pulmonary pressures during exercise, which force fluid out of pulmonary capillaries. Specific airway diseases in horses, especially those associated with hypoxaemia, hypercapnoea and acidosis may influence pulmonary haemodynamics and pulmonary interstitial fluid equilibrium. OBJECTIVES: This study was designed to determine fluid flux (J(V-A) l/min) across the lung in exercising horses treated chronically with acetazolamide. METHODS: Six horses were exercised on a treadmill until fatigue without (Con) and with chronic carbonic anhydrase (CA) inhibition (AczTr) and associated hypercapnoea and acidosis. Carbonic anhydrase inhibition was achieved with administration of acetazolamide (Acz). Arterial and mixed venous blood were sampled, and VCO2 and VO2 measured. Blood volume changes across the lung (deltaBV%) were calculated from changes in plasma protein, haemoglobin and packed cell volume (PCV). Cardiac output (Q) was calculated using Fick principle. J(V-A) across the alveolar-capillary barrier was then quantified based on Q and deltaBV. Variables were analysed using 2-way repeated-measures ANOVA (P<0.05). A significant F ratio was further analysed using Tukey post hoc analysis. RESULTS: Treatment had a significant effect on J(V-A) (P = 0.002). At rest there was no J(V-A) in Con (0.63 +/- 0.6 l/min) and AczTr (0.84 +/- 0.3 l/min). During exercise Con fluid moved from the pulmonary circulation into the pulmonary interstitium (mean +/- s.e. J(V-A) 9.4 +/- 2.4 l/min). This was different from AczTr (mean +/- s.e. J(V-A) 1.8 +/- 1.9 l/min), where no transvascular fluxes from pulmonary circulation were present during exercise (P = 0.008). CONCLUSIONS: Chronic Acz treatment with associated hypercapnoea and acidosis affects J(V-A) in lungs of exercising horses. Lung fluid dynamics adapted to hypercapnoea and acidosis with reduction of fluid flow from the pulmonary circulation. POTENTIAL RELEVANCE: The current data provide comprehensive evidence of in vivo fluid homeostasis in lungs of exercising horses without and with CA inhibition. PMID- 17402476 TI - Effect of corticosteroid and bronchodilator therapy on bronchoalveolar lavage cytology following intrapulmonary blood inoculation. AB - REASON FOR PERFORMING STUDY: Intrapulmonary blood causes an inflammatory response. Modifying this inflammatory response may be useful in the treatment of exercise induced pulmonary haemorrhage (EIPH). OBJECTIVES: To evaluate the efficacy of 3 treatments, an oral bronchodilator, inhaled steroid and oral steroid in modifying the cellular response to intrapulmonary blood inoculation. MATERIALS AND METHODS: Experimental design was a randomised block divided into 4 experimental segments of 2 weeks. Eight normal horses were randomly assigned to pairs and treatments. Autologous blood was inoculated into the lungs on 4 occasions over an 8 week period. For each treatment, a bronchoalveolar lavage (BAL) sample was collected on Day 0 and then 2 randomly selected segmental bronchi (one in each lung) were inoculated with blood (50 ml). BAL fluid was collected from one inoculated bronchus on Day 3 and from the other on Day 10. From BAL samples, total RBC and total and differential WBC were determined. Results for percentages and absolute cell counts were expressed as mean +/- s.e. and analysed using one, 2 and 3 way analysis of variance. RESULTS: All treatments significantly reduced the number of erythrophages at Day 3 (P = 0.02) Treatment with oral prednisolone resulted in a significantly higher percentage of macrophages (P = 0.03) and also reduced the relative and absolute number of haemosiderophages in BAL compared to the other 2 treatments at Day 10 (P = 0.07). CONCLUSION: We concluded that treatments altered the dynamics of cell populations in BAL following blood inoculation. Whether this alters the evolution of lung lesions following episodes of EIPH and indeed whether there are benefits for equine lungs following episodes of EIPH, remain to be determined. POTENTIAL RELEVANCE: Future studies into treatments for EIPH may benefit from incorporating a variety of dose rates for oral prednisolone and a longer period of evaluation. Such studies may also benefit from the use of cell activation markers. The concomitant use of pulmonary function testing and cytology in the field may assist in ascertaining whether treatments influence the performance and welfare of athletic horses subsequent to episodes of EIPH. PMID- 17402477 TI - Correlations between exercising arterial blood gas values, tracheal wash findings and upper respiratory tract abnormalities in horses presented for poor performance. AB - REASONS FOR PERFORMING STUDY: There are limited data on the correlations between arterial blood gas (ABG) values, tracheal wash (TW) cytology and upper respiratory tract (URT) abnormalities. OBJECTIVES: To identify horses with abnormal exercising ABG, and compare the proportions of horses with abnormal ABG and TW cytology, mucus or URT dysfunction with those with normal ABG results and abnormal TW cytology, mucus or URT dysfunction. METHODS: Medical records of 813 horses presenting to the treadmill facility that had a complete treadmill examination, including ABG analysis, TW and URT endoscopy were selected. Diagnoses, ABG results, TW cytology and URT endoscopy were compared. RESULTS: Two hundred and eleven horses met the study criteria of a complete treadmill examination and could have ABG evaluated. There were no significant differences in the age distribution of horses having normal and abnormal ABG or upper respiratory tract (URT) examinations. There was a significantly higher percentage of geldings with abnormal ABG analysis. In the horses with abnormal URT examinations, there were no differences in the proportion of horses having mucus vs. no mucus. However, in the horses with normal URT, there were a higher percentage of horses with visible mucus in the group with abnormal ABG analysis. The majority of horses had abnormal TW cytology and evidence of prior EIPH, with no differences in proportions between the groups. CONCLUSIONS: Because such a large percentage of horses had evidence of inflammation and/or evidence of prior EIPH on TW cytology, it was not possible to determine the effect of these findings on gas exchange. Mucus was present in a larger percentage of cases with abnormal ABG analysis and normal URT examinations, suggesting that the presence of mucus may affect gas exchange. Standardbreds may be more likely to have abnormal gas exchange than Thoroughbreds. A larger number of horses is needed to determine the significance of these findings. POTENTIAL RELEVANCE: Abnormal TW cytology and endoscopic visualised mucus may contribute to impairment of gas exchange, but they do not specifically predict abnormal ABG analysis. PMID- 17402478 TI - Prevalence of inflammatory airway disease in national hunt horses referred for investigation of poor athletic performance. AB - REASONS FOR PERFORMING STUDY: Inflammatory airway disease (IAD) is thought to be an important cause of poor performance in young Thoroughbred racehorses. However, little study has been made of IAD in older National Hunt (NH) horses. OBJECTIVES: To determine the prevalence of IAD in NH racehorses referred for investigation of poor athletic performance and identify some of the risk factors that may be associated with IAD in this group of horses. METHODS: Tracheal mucus was graded, and tracheal wash (TW) and bronchoalveolar lavage (BAL) performed after treadmill exercise in 91 NH horses referred to the University of Bristol. Comparisons were made between the different methods for diagnosing IAD and potential risk factors investigated. RESULTS: Tracheal mucus was observed in 68% and lower airway inflammation identified in 70% of horses. There was poor agreement between TW and BAL techniques for a diagnosis of IAD. The prevalence of increased proportions of neutrophils in TW was 40% compared with 59% in BAL. There was a significant association between presence of tracheal mucus and increased neutrophils in TW but not between tracheal mucus and BAL cytology. No significant association between IAD and age, EIPH or URT obstruction was observed. CONCLUSIONS: Inflammatory airway disease was a common finding in NH horses referred for investigation of poor performance. In contrast to studies in younger, flat racehorses the prevalence of disease did not decrease with increasing age. POTENTIAL RELEVANCE: Horses of all ages presented for investigation of poor performance require a thorough clinical investigation of the lower airways. The collection of both TW and BAL samples is indicated in order to confirm a diagnosis of IAD. PMID- 17402479 TI - Cold air-induced late-phase bronchoconstriction in horses. AB - REASON FOR PERFORMING STUDY: Inspired air is warmed to body temperature and fully humidified by the upper airway mucosa under normal resting conditions. This conditioning process may not be completed by the upper airways during conditions of increased minute ventilation or when the inspired air is unusually cold, resulting in cooling and desiccation of lower respiratory surfaces. Excess heat and water loss from intrapulmonary airways is believed to be the provocative stimulus for exercise-induced bronchoconstriction (occurring immediately after exercise) and associated late phase airway obstruction (occurring a few hours after exercise). HYPOTHESIS: Exercise while breathing cold air results in airway obstruction in horses. METHODS: Eight healthy horses performed a 15 min submaximal exercise challenge in a random crossover design. Independent variable was inspired air temperature during the challenge (25 or -5 degrees C). The dependent variables were total respiratory impedance, resistance, and reactance at 5, 24 and 48 h post exercise challenge, expressed as a percentage of the prechallenge baseline. RESULTS: No significant effect of inspired air temperature was found on any respiratory mechanical parameter 5 h after exercise challenge. However, cold inspired air was associated with higher respiratory impedance and resistance 48 h after the exercise challenges. CONCLUSIONS: These findings support the hypothesis that submaximal exercise while breathing subfreezing air can adversely affect respiratory mechanical properties in normal horses. However, the timecourse for development of abnormal respiratory mechanical properties is longer than that reported in other mammals. CLINICAL RELEVANCE: Exercise in cold weather may be a common cause of lower airway disease in horses. PMID- 17402480 TI - Videoendoscopic evaluation of the upper respiratory tract in 93 sport horses during exercise testing on a high-speed treadmill. AB - REASONS FOR PERFORMING STUDY: Videoendoscopy of the upper respiratory tract (URT) during high-speed treadmill exercise has proved to be invaluable in the assessment of URT dysfunction in racehorses. However, very little information exists regarding dynamic airway collapse in other sport horses used in nonracing equestrian disciplines. OBJECTIVES: To evaluate the videoendoscopic findings at rest and during exercise in a mixed population of sport horses referred for investigation of poor athletic performance and/or abnormal respiratory noise. METHODS: Videoendoscopy of the upper airway was performed at rest and during high speed treadmill exercise in 93 horses. RESULTS: Dynamic airway obstructions were diagnosed in 77% of horses and were frequently complex in nature. The most common forms of dynamic collapse included soft palate dysfunction (54%), dynamic laryngeal collapse (38%), axial deviation of the aryepiglottic folds (24%) and pharyngeal wall collapse (18%). In the majority of horses, no obvious abnormalities were identified at rest. Enforced poll flexion was found to be a contributing factor in 24% of cases. CONCLUSIONS: Dynamic obstructions of the URT were a common cause of poor performance and/or abnormal respiratory noise in sport horses referred for investigation of performance problems. POTENTIAL RELEVANCE: This study highlights the importance of videoendoscopic evaluation of the URT during exercise in horses utilised for equestrian sports where exercise during competition is submaximal in nature. PMID- 17402481 TI - Dynamic pharyngeal collapse in racehorses. AB - REASON FOR PERFORMING STUDY: Dynamic pharyngeal collapse (PC) is a condition seen in racehorses that can be career-ending. OBJECTIVES: To characterise and grade PC and describe the effects of PC on athletic performance. METHODS: Medical records were reviewed for 828 horses, of which 49 (6%) records were identified as horses with a primary diagnosis of PC. Tapes of video-endoscopy of the pharynx during exercise were reviewed. Each video recording was assigned a grade (0-4) reflecting the degree of PC and a classification for severity of upper airway obstruction. Earnings per race prior to diagnosis of PC were compared to earnings per race after diagnosis of PC for all horses, as well as performance index (PI). Available exercising arterial blood gases were reviewed for horses with PC. RESULTS: There were 35 (80%) Thoroughbreds (TB), and 9 (20%) Standardbreds (STD). 32 (73%) had a history of making an upper respiratory noise. 4 (9%) grade 1 PC, 8 (18%) grade 2 PC, 26 (59%) grade 3 PC, and 6 (14%) grade 4 PC. Seven (16%) horses were classified as mild PC, 18 (41%) as low-moderate PC, 14 (32%) as high moderate PC, and 5 (11%) as severe PC. Of 30 horses 11 had abnormally decreased PaO2 and 8 horses had abnormally elevated PaCO2. A significant decrease was found in earnings per race prediagnosis when compared to post diagnosis earnings per race in horses > or =4 years of age (P = 0.003). A significant decrease was also observed for earnings per race prediagnosis when compared to post diagnosis earnings per race in horses with grade 3 PC (P = 0.03) No significant differences were observed in PI before or after diagnosis of PC. CONCLUSIONS: There was a trend for PC to be observed in more TB than STD, and more males than females compared to the general hospital population. Horses with PC significant had decreases in arterial oxygenation. Racing records after a diagnosis of PC in all horses > or = 4 years of age suggesting that older horses have a guarded prognosis for continued success. POTENTIAL RELEVANCE: This study provides a classification system for dynamic pharyngeal collapse and suggests that older racehorses (> or = 4 years of age) diagnosed with PC and all horses with grade 3 PC have a poor prognosis for return to previous level of performance. PMID- 17402482 TI - Haematological and respiratory gas changes in horses and mules exercised at altitude (3800 m). AB - REASON FOR PERFORMING STUDY: Despite the common use of equids as visitors to high altitude mountainous environments, there are a paucity of carefully orchestrated scientific approaches. Further, again as a function of a common perceived advantage of mules over horses in these similar environments there are needs for controlled comparisons between these 2 equids. OBJECTIVE: To measure haematological and respiratory function in horses and mules at low altitude (225 m), at rest and post exercise. In addition the rate and magnitude of these changes were followed over a 13 day period at high altitude (3800 m) to contrast acclimatisation. METHODS: Resting and exercise venous blood samples (1 min post exercise) were obtained from 6 horses and 5 mules housed at 225 m (LA) and then transported to 3800 m (HA) for 13 days. The standardised exercise tests at both LA and HA consisted of trotting (3.0 m/sec) up an incline (6%) for 2 km. Data were analysed with repeated measures ANOVA (comparison of altitude acclimatisation and species) for changes in haematological and respiratory gases. RESULTS: At low altitude, no group differences were found with both resting (P = 0.69) and exercising (P = 0.74) heart rates. Resting PCV was 8% lower in the mules (P = 0.02) and 20% lower during exercise (P = 0.02). Horses had significantly higher 2,3-diphosphoglycerate (2,3-DPG)/g Hb at both rest (P = 0.003) and exercise (P = 0.03). Exercise at HA increased PCV (P = 0.03) in both groups, but the increase was attenuated in the mules compared to horses. The increase with 2,3-DPG/g Hb was expressed at HA in both groups (P = 0.001) and was also attenuated in mules (P = 0.03). Both groups were alkalotic compared to LA (P = 0.001), and there were no group differences (P = 0.95). CONCLUSION: Of the variables measured, the most notable distinction between species was identified for only PCV and 2,3-DPG with both higher in horses, at both LA and HA. While the attenuated response of PCV in mules for the same exercise might argue for an improved adaptation to altitude, the lower 2,3-DPG might not. Other variables during the exercise bout were not different between species. PMID- 17402483 TI - Validation of a portable equine metabolic measurement system. AB - REASONS FOR PERFORMING STUDY: In equine sports medicine, VO2 has been measured exclusively with stationary systems, in laboratories equipped with a treadmill. Measurement during exercise in field conditions has not previously been reported because of the lack of portable equipment designed for horses. OBJECTIVES: A commercially available portable metabolic measurement system, based on breath-to breath gas analysis and flow spirometry, was adapted to the horse's physiology and morphology (Cosmed K4b2 and Equimask) and its validity tested by (1) repeatability of the measures and (2) comparing metabolic data to those obtained by a reference method (RM). METHODS: To test the reproducibility of the measurements, 5 healthy saddle horses were subjected twice at 2 day intervals to a similar submaximal standardised incremental exercise test on a treadmill. The same horses performed twice at one week interval an incremental treadmill test to fatigue: the oxygen consumption and ventilation were measured once with the K4b2 system and once with the RM. The metabolic and ventilatory data obtained with both systems were compared. RESULTS: There was a good reproducibility of the metabolic measurements obtained by the K4b2 system at any workload. The VO2 obtained by both systems at any workload was not significantly different. However, the K4b2 expired fraction in CO2 (FETCO2) and carbon dioxide production (VCO2) were significantly lower at high and at maximal workloads. As a consequence, the values of the respiratory exchange ratio were too low and incompatible with normal physiological values. CONCLUSIONS: The good reproducibility of the metabolic and ventilatory measurements and the fact that the VO2 measurements at any workload were similar to the data obtained with the reference method suggested that this system may be used for comparison of repeated VO2 measurements in practical field conditions. POTENTIAL RELEVANCE: The K4b2 system could be used to improve knowledge of the energetic cost in different equine sports disciplines and offer the opportunity to undertake performance tests with genuine track conditions, on ridden or harnessed horses, rather than under laboratory conditions. PMID- 17402484 TI - Factors influencing variation in locomotor-respiratory coupling in standardbred trotters in the field. AB - REASONS FOR PERFORMING STUDY: A close relationship between limb and respiratory rhythms has been shown in clinically healthy galloping horses due to mechanical constraints in the thoracic region. This synchronisation leads to a 1/1 ratio between stride frequency (SF) and respiratory frequency (RF) during galloping. Very little is known about locomotor-respiratory coupling (LRC) during fast trot. OBJECTIVES: To investigate stride and respiratory rates during a standardised exercise test on the track in Standardbred Trotters. METHODS: Forty-four French trotters age 2-10 years performed a standardised exercise test consisting of three 3 min steps at constant speeds. Speed (V) of exercise varied between 500 and 750 m/min. Variables measured during exercise: SF, heart rate, RF using a microphone between the nostrils, blood lactate concentration. Physiological variables V4 and V200 were calculated and race performance index recorded. RESULTS: There was no age-related difference in RF or in LRC. Two LRC strategies were found: around 1/1 ratio and 3/2 ratio between SF and RF, respectively. A relationship between SF/RF ratio and physiological parameters showed a higher LRC in good performers because of a lower RF during submaximal and maximal exercise. CONCLUSION: RF is significantly lower and LRC significantly higher in good compared to poor performers. POTENTIAL RELEVANCE: Investigation is required to ensure that the breathing techniques of trotters may be used in the selection process of racehorses. PMID- 17402485 TI - Observations on respiratory flow strategies during and after intense treadmill exercise to fatigue in thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: Locomotor-respiratory coupled (LRC) breath types are a feature of galloping exercise in horses. Differences in breath type have been demonstrated during exercise in particular the 'big respiratory cycle' (BRC) and 'flow hesitation'. To investigate breath types during recovery and quantitatively investigate BRCs during exercise to understanding the mechanism driving BRCs. OBJECTIVES: To investigate the occurrence of different breath types during and after intense treadmill exercise, and test the hypothesis that large breaths (BRCs) were a function of respiratory frequency. METHODS: Six trained and clinically normal Thoroughbred horses were exercise tested on a treadmill (slope 10%). Breath-by-breath pulmonary ventilation was measured continuously during exercise and recovery using a Quadflow mask. RESULTS: Five different breath types were identified, and classified as normal monophasic, normal biphasic, deglutition, effort pause, or large breaths. Exercising at 10 m/sec, the number of large breaths was significantly related to Rf (r = -0.86, P = 0.03). During 120 sec after exercise there were 2 distinct populations of breaths, large and normal monophasic. CONCLUSIONS: BRC type breaths are a normal feature of ventilation during and after intense exercise. In recovery there are two distinct breath populations. During exercise BRC frequency is inversely associated with respiratory frequency and highly dependant on the individual horse. From intense exercise to recovery, high flow rates and LRC limited tidal volumes are replaced by high tidal volumes and progressively decreasing flow rates. There is a temporal association between BRC occurrence and PECO2. POTENTIAL RELEVANCE: Breath types and the physiological mechanism for driving each type is important in the clinical interpretation of respiratory disease or dysfunction. The demonstration of BRC association with PECO2 may help understand the driving mechanism for the BRC. In pulmonary function testing, breath type is important in quantitative results. The demonstration that high tidal flows with limited tidal volumes during intense exercise being replaced by high tidal volumes and progressively decreasing flows in recovery has potential clinical relevance. PMID- 17402486 TI - Oxygen consumption (VO2) during trotting on a 10% decline. AB - REASONS FOR PERFORMING STUDY: Although there have been reports of oxygen consumption measurements of horses running on the level and incline, there are no measurements during decline locomotion. This may be due, in part, to the potential for muscle damage produced by eccentric contractions. In man, running on a 10% decline, VO2 decreased by 35% and stride frequency (SF) decreased by 3% when compared to level locomotion. HYPOTHESIS: The rate of O2 consumption and SF would be decreased in horses on a 10% decline when compared to the level. METHODS: Six horses (average 467 +/- 68 kg) were acclimated to trotting on the level and decline prior to data collection. VO2 under moderate conditions was measured (using open flow respirometry) during trotting between 2.25 and 4.0 m/sec (at 0.25 m/sec increments) on a treadmill on the level and declined 10%. Stride frequencies were counted manually. RESULTS: VO2 decreased (P<0.009) on the decline by an average of 45% (range 42-47%), and SF was 2.7% slower. The speed at which the minimum Cost of Transport occurs on the decline was faster than on the level. SF was reduced on the decline. No evidence of muscle soreness was noted in response to the downhill running. CONCLUSIONS AND POTENTIAL RELEVANCE: Downhill trotting, eccentric exercise, can be done safely in the horse and requires almost half the energetic costs as trotting on the level. It is not known whether this is the optimum downhill gradient or if the horse adjusts its preferred speed to accommodate downhill trotting. PMID- 17402487 TI - Effects of dietary energy source and physical conditioning on insulin sensitivity and glucose tolerance in standardbred horses. AB - REASONS FOR PERFORMING STUDY: There is evidence that adaptation to diets rich in nonstructural carbohydrates (NSC) contributes to the development of insulin resistance in horses. To date, however, no study in horses has examined the effects of physical conditioning on diet-associated alterations in insulin sensitivity and glucose tolerance. OBJECTIVES: To examine the effects of adaptation to concentrate feeds rich in NSC or fat on insulin sensitivity and glucose tolerance in horses, both in the sedentary state and after a subsequent period of physical conditioning. METHODS: Fourteen mature Standardbred horses underwent both a euglycaemic-hyperinsulinaemic clamp (EHC) and oral glucose tolerance test (OGTT) after each of the following phases: Baseline - fed only forage cubes for 3 weeks; Diet horses were randomly assigned to receive either a high NSC (S) concentrate or a high fat concentrate (F) with forage cubes for 6 weeks; and Diet x Exercise - horses remained on the assigned ration and underwent a 7 week period of physical conditioning. An incremental exercise test was performed before, and after, the Diet x Exercise phase for measurement of the peak rate of oxygen consumption (VO2peak). RESULTS: In both diet groups, there was an approximately 10% increase in mean VO2peak after physical conditioning. The mean rate of glucose disposal (M) per unit of serum insulin (I) during the EHC [M/I ratio] in S horses was 30% lower (P<0.05) in the Diet phase when compared to Baseline, but not different from Baseline after physical conditioning. The S diet also resulted in a greater (P<0.05) OGTT insulin response (area under the insulin vs. time curve, AUC(INS)) in both Diet and Diet x Exercise phases when compared to Baseline. In F, insulin sensitivity (mean M/I ratio) and glucose tolerance were unchanged during the study. CONCLUSIONS AND POTENTIAL RELEVANCE: Feeding a diet rich in NSC for 6 weeks resulted in decreased insulin sensitivity and impaired glucose tolerance. Physical conditioning lessened the effects of the high NSC diet on insulin sensitivity, as evidenced by the return to baseline M/I, but did not mitigate the impaired glucose tolerance. Decreased insulin sensitivity has been implicated in the development of obesity and laminitis in horses and the present findings provide support for avoidance of concentrates with high NSC in the dietary management of horses at risk for the development of these conditions. PMID- 17402488 TI - Effects of dietary glycaemic response after exercise on blood concentrations of substrates used indirectly for muscle glycogenesis. AB - REASONS FOR PERFORMING STUDY: Exercise depletes muscle glycogen stores, which could subsequently impair performance. Muscle glycogen replenishment is determined by substrate availability. OBJECTIVES: To study the effects of feeding meals of varying glycaemic responses on blood concentrations of substrates used for glycogenesis in horses with exercise-induced glycogen depletion. METHODS: In a 3-way crossover study, 7 horses received each of 3 isocaloric diets for 72 h after undergoing glycogen-depleting exercise: 1) a high soluble-carbohydrate diet, which induced a high-glycaemic (HGI) response; 2 and 3) a low soluble carbohydrate or a mixed soluble-carbohydrate diet (control group), which both induced a similar low-to-moderate glycaemic (LGI) response. Muscle biopsies and venous samples were collected before and up to 72 h after exercise. RESULTS: Feeding HGI diet resulted in a higher (P<0.001) rate of muscle glycogenesis over 72 h compared to LGI diets. Plasma glycerol, triglyceride, lactate, serum NEFA and total protein concentrations, and haematocrit were significantly (P<0.001) higher after compared to before exercise. Whereas no significant overall dietary effect was observed on these metabolites over 72 h, there was a tendency for glycerol, NEFA and triglyceride concentrations to be lower for LGI compared to HGI diets over 6 h after exercise (P<0.05; 1, 6 and 4-6 h after exercise, respectively). CONCLUSIONS: These data suggest that horses fed LGI meals after exercise had limited lipid utilisation without any significant shift of substrate utilisation toward gluconeogenesis, which could have contributed to the slower rate of muscle glycogenesis compared to horses fed HGI diets. PMID- 17402489 TI - Route of carbohydrate administration affects early post exercise muscle glycogen storage in horses. AB - REASONS FOR PERFORMING STUDY: No studies in horses have examined the effect of route of carbohydrate (glucose) administration on the rate of muscle glycogen storage following glycogen-depleting exercise. HYPOTHESIS: Glucose delivery from the gastrointestinal tract limits the rate of muscle glycogen storage following glycogen-depleting exercise. METHODS: In a crossover design, 7 fit horses completed treadmill exercise (EX) on 3 occasions to deplete muscle glycogen by approximately 50%. After EX horses received: 1) i.v. glucose infusion (IV; 0.5 g/kg bwt/h for 6 h), 2) oral glucose boluses (OR; 1 g/kg bwt at 0, 2 and 4 h post EX) or 3) no glucose supplementation (CON). Blood samples for measurement of glucose and insulin concentrations were collected before EX and during the 6 h treatment period. Muscle biopsies for measurement of muscle glycogen content (GLY) and glycogen synthase (GS) activity were taken before and after exercise and at 3 and 6 h. RESULTS: Mean plasma glucose concentrations were significantly higher in IV and OR than in CON throughout treatment. The average serum insulin responses in IV and OR treatments were also significantly greater than in CON. After EX, GLY was not different among the 3 treatments. However, glycogen storage rates were significantly higher in IV than in CON and OR during the first 3 h and second 3 h of recovery, and GLY was significantly higher in IV than in OR and CON at 6 h of recovery. GS activity was significantly higher in IV than in OR and CON at 3 h of recovery. CONCLUSIONS: Muscle glycogen storage in horses during a 6 h period after exercise was enhanced by i.v. glucose administration (3 g/kg) but not by an equivalent glucose dose administered per os. While oral administration of glucose achieved a level of hyperglycaemia and hyperinsulinaemia that markedly accelerates glycogen storage in other species, the rate of glycogen storage following oral supplementation was not different to control conditions. POTENTIAL RELEVANCE: Glucose supplementation via the i.v. route should be considered when rapid replenishment of muscle glycogen stores is desired. PMID- 17402490 TI - Interval exercise alters feed intake as well as leptin and ghrelin concentrations in standardbred mares. AB - REASONS FOR PERFORMING STUDY: Horses in training tend to become inappetant; however, the mechanism responsible for this training-induced inappetance is not known. HYPOTHESIS: Training and/or ulcers alter the feed intake (FI) and hormonal and/or biochemical (active ghrelin, leptin, glucose, insulin and cortisol) responses to acute high intensity exercise. METHODS: Eight Standardbred mares underwent 3 interval exercise tests (IET) and 3 parallel control tests (CON) before (IET1) and after 8 weeks of training (IET2) and after treatment for gastric ulcers (IET3). Plasma samples were taken before (0 min), during (last 10 sec of velocities eliciting 40, 100 and 20% VO2max), and after (30 min, 60 min, 24 h) exercise (EX) or CON tests for RIA and colorimetric measurement of the concentrations of the above parameters. Samples were also collected before and after feeding. Horses were trained at a work intensity of 70% HRmax for 30 min/day, 5 days per week with FI measured daily. RESULTS: There were no changes (P>0.05) in any variable during the parallel control trials. However, there was a mismatch between FI and digestible energy (DE) requirements (P<0.05) with EX horses not meeting their DE requirements during the post training IETs. During all IETs, ghrelin, glucose and cortisol increased (P<0.05) during EX. Leptin only increased (P<0.05) during EX in the post training IETs. Insulin remained low during EX, but increased (P<0.05) post EX. CONCLUSION: High intensity exercise appeared to be associated with decreases in FI and alterations of leptin and ghrelin. POTENTIAL RELEVANCE: More research is needed to determine if there is a relationship between alterations of these hormones and changes in FI in horses that lose weight while in training. PMID- 17402491 TI - Study of gelatin supplemented diet on amino acid homeostasis in the horse. AB - REASONS FOR PERFORMING STUDY: Gelatin supplementation is a common measure in an attempt to assist cartilage repair, but little scientific evidence exists to support its efficacy. OBJECTIVES: To investigate the effects of gelatin administration on post prandial homeostasis. METHODS: Twelve Standardbred horses (mean 404 kg bwt) were fed a hay-concentrate diet supplemented by soy bean meal and oil (control [C], n = 6) or with the addition of 60 g gelatin/day (G, n = 6). The horses were trained by an alternate order of interval and prolonged exercise every second day. The velocities of the treadmill corresponding to 2 and 10 mmol lactate/l blood were derived from lactate curves during a standardised exercise test at the start and middle of the 64 day training period. Blood samples for amino acid analysis were obtained weekly at rest (2 h post prandial). In the second part of the training period, a post prandial sampling was conducted on a day without exercise (prior feeding up to 8 h post prandial). Plasma free amino acids (AA) were determined by HPLC. RESULTS: The change from pre- to the training diet induced an increase in many AA during the total training period. At rest free glycine and proline in blood increased with gelatin supplementation during 7 days after the start of supplementation. The AA in plasma showed a post prandial curve with peak concentrations 2-3 h after feeding. Significant post prandial effects of gelatin intake were detectable for glycine, proline and arginine. CONCLUSIONS: The AA from gelatin are absorbed quickly and become available for AA metabolism. POTENTIAL RELEVANCE: It is evident that in the horse, gelatin influences the homeostasis of those amino acids required for cartilage synthesis. Further research is needed to elucidate the utilisation of those amino acids for the prevention or repair of cartilage damage. PMID- 17402492 TI - Effect of amino acid and glucose administration following exercise on the turnover of muscle protein in the hindlimb femoral region of thoroughbreds. AB - REASONS FOR PERFORMING STUDY: In man, muscle protein synthesis is accelerated by administering amino acids (AA) and glucose (Glu), because increased availability of amino acids and increased insulin secretion, is known to have a protein anabolic effect. However, in the horse, the effect on muscle hypertrophy of such nutrition management following exercise is unknown. OBJECTIVES: To determine the effect of AA and Glu administration following exercise on muscle protein turnover in horses. We hypothesise that administration of AA and Glu after exercise effects muscle hypertrophy in horses, as already shown in man and other animals. METHODS: Measurements of the rate of synthesis (Rs) and rate of degradation (Rd) of muscle protein in the hindlimb femoral region of thoroughbred horses were conducted using the isotope dilution method to assess the differences between the artery and iliac vein. Six adult Thoroughbreds received a continuous infusion of L-[ring-2H5]- phenylalanine during the study, the stable period for plasma isotope concentrations (60 min), resting periods (60 min), treadmill exercise (15 min) and recovery period (240 min). All horses were given 4 solutions (saline [Cont], 10% AA [10-AA], 10% Glu [10-Glu] and a mixture with 10% AA and 10% Glu [10-Mix]) over 120 min after exercise, and the Rs and Rd of muscle protein in the hindlimb measured. RESULTS: The average Rs during the 75-120 min following administration of 10-Mix was significantly greater than for the other solutions (P<0.05). The second most effective solution was 10-AA, and there was no change in Rs after 10-Glu. CONCLUSIONS: Administration of AA following exercise accelerated Rs in the hindlimb femoral region, and this effect was enhanced when combined with glucose, because of increasing insulin secretion or a decreased requirement for AA for energy. POTENTIAL RELEVANCE: Further studies are required regarding the effect on muscle hypertrophy of supplementing amino acids and glucose in the feed of exercising horses. PMID- 17402493 TI - Oral vitamin E supplementation on oxidative stress, vitamin and antioxidant status in intensely exercised horses. AB - REASONS FOR PERFORMING STUDY: Vitamin E is the most commonly supplemented antioxidant in horses; however, previous research is not conclusive as to the recommended level for exercising horses. OBJECTIVE: To evaluate the effects of 3 levels of vitamin E supplementation on oxidative stress and vitamin/antioxidant status in intensely exercised horses to determine the optimal level of vitamin E supplementation. METHODS: Twelve unfit Standardbreds were divided into 3 groups, supplemented orally with 0 (CON), 5000 (MOD), or 10,000 (HI) iu/day of DL-alpha tocopheryl acetate. The 3 x 3 Latin square design consisted of three 4 week supplementation periods with 4 week wash out periods between. After each period, horses underwent a treadmill interval exercise test. Blood samples were collected and heart rate (HR) measured before, during and after exercise. Data were analysed using ANOVA with repeated measures in SAS. RESULTS: The CON group had lower HR throughout the test compared to the MOD and HI groups (P<0.05). There was an increase in plasma retinol (RET), beta-carotene (BC), red blood cell total glutathione and glutathione peroxidase with exercise (P<0.05), but all groups returned to baseline after 24 h. Plasma alpha-tocopherol (TOC) increased from baseline with exercise (P<0.0001) in all groups; treatment differences were observed at 24 h (P<0.05). The HI and CON groups had lower BC compared to the MOD group (P = 0.05). CONCLUSIONS: Horses supplemented with vitamin E, at nearly 10 times the 1989 NRC recommended level, did not experience lower oxidative stress compared to control horses. Additionally, lower plasma BC levels observed in the HI group, which may indicate that vitamin E has an inhibitory effect on BC metabolism. POTENTIAL RELEVANCE: Supplementation above control levels is not more beneficial to oxidative stress and antioxidant status in intensely exercising horses; indeed, levels 10 times in excess may be detrimental to BC and should be avoided. PMID- 17402494 TI - Double blind investigation of the effects of oral supplementation of combined glucosamine hydrochloride (GHCL) and chondroitin sulphate (CS) on stride characteristics of veteran horses. AB - REASONS FOR PERFORMING STUDY: Oral chondroprotective supplements are commercially popular for veteran (and other athletic or arthritic) horses prone to joint degeneration, yet lack conclusive scientific support. OBJECTIVES: To quantify the effects of an oral joint supplement (combination glucosamine hydrochloride (GHCL), chondroitin sulphate (CS) and N-acetyl-D-glucosamine) in vivo on stride parameters of veteran horses. METHODS: Twenty veteran horses were randomly assigned to a treatment (n = 15) or placebo group (n = 5). Pre-treatment gait characteristics were recorded at trot using digital video footage (50 Hz). The range of joint motion, stride length, and swing and stance duration were assessed using 2-dimensional motion analysis. Treatment (or placebo) was administered daily for 12 weeks at the manufacturer's recommended dosage. Gait was reassessed every 4 weeks using the pre-treatment protocol. Double blind procedure was implemented throughout. Relationships between variables were analysed using General Linear Model. RESULTS: Differences occurred in the treated horses by week 8. Range of joint motion increased significantly in the elbow (P<0.05), stifle and hind fetlock (P<0.01). Stride length increased significantly (P<0.05) with treatment. Swing duration was significantly increased at week 12 (P<0.05), whilst stance duration remained constant. CONCLUSION: The oral chondroprotective offered symptomatic relief to veteran horses, evidenced by improved stride characteristics. POTENTIAL RELEVANCE: Oral GHCL and CS supplementation may improve welfare by alleviating symptoms of degenerative joint disease. PMID- 17402495 TI - Exercise response, metabolism at rest and digestibility in athletic horses fed high-fat oats. AB - REASON FOR PERFORMING STUDY: High starch intakes increase the risk for metabolic disorders and therefore alternative feedstuffs are of interest. High-fat oat varieties have a lower starch and higher energy content than regular oats and may therefore be useful in this context. HYPOTHESIS: Feeding high fat oats causes no adverse effects on the response to exercise and that the total amount of oats offered could be reduced compared to feeding with regular oats. METHODS: Twelve Standardbred trotters were fed regular oats (diet C), high-fat oats (F), and a mixture (50:50) of C and F (M), together with haylage (30:70), in a Latin square design trial. High-fat oats replaced regular oats in a 0.9 to 1.0 ratio in diets F and M. On Day 18 in each 21 day experimental period, horses were subjected to a standardised near-maximal treadmill exercise test with collection of blood samples and muscle biopsies before and after exercise. This was followed by a 3 day period of total collection of faeces and urine. RESULTS: There were no significant effects of dietary treatments on bodyweight, heart rate, plasma lactate and glucose, or on muscle glycogen and lactate concentrations following exercise. However, plasma insulin was reduced during exercise on diets F and M compared to diet C. The total tract digestibility of dry matter, fat, protein, NDF and organic matter were higher for diet F than for diet C. CONCLUSION: High fat oats can replace regular oats in the diet of athletic horses without any adverse effects on metabolism and exercise response. POTENTIAL RELEVANCE: Due to the high energy content and a high digestibility of dietary components in high fat oats the daily allowance of oats can be reduced and thus the intake of starch. PMID- 17402496 TI - Glucose dynamics during exercise: dietary energy sources affect minimal model parameters in trained Arabian geldings during endurance exercise. AB - REASONS FOR PERFORMING STUDY: Glucose regulation is critical for health and exercise performance. OBJECTIVES: To quantify the effects of exercise and diet on insulin sensitivity (SI), glucose effectiveness (Sg), acute insulin response to glucose (AIRg) and disposition index (DI) in horses. METHODS: This study applied the minimal model of glucose and insulin dynamics to exercise-trained Arabian geldings during rest or constant moderate-intensity exercise after 8 weeks adaptation to feeds high in sugar and starch (SS, n = 6) or fat and fibre (FF, n = 6). Horses underwent 2 frequently sampled i.v. glucose tolerance tests (FSIGT). For both tests, a resting basal sample was collected, followed by an i.v. dose of 600 mg/kg bwt glucose defining 0 min of the test. Insulin (0.01 iu/kg bwt) was administered 20 min post glucose for each test. Resting horses were sampled for 240 min. The exercise FSIGT began after each horse had warmed-up for 25 min on the treadmill at which point they had reached the speed representing 60% of their predetermined lactate breakpoint maintained for the rest of the FSIGT. Exercising horses were sampled identically to rest, but for only 150 min post glucose. RESULTS: Exercise increased (P<0.008) SI, Sg and DI and decreased AIRg in all horses. Overall, horses adapted to FF tended to have higher SI (P = 0.070) and DI (P = 0.058). During exercise, FF horses tended to have higher (P< or =0.085) SI and DI, than SS horses and these variables tended to be increased more (P< or =0.075) by exercise in FF horses than SS horses. CONCLUSIONS: Insulin and glucose dynamics adjust during exercise, increasing plasma glucose uptake, presumably to meet demand by contracting skeletal muscle. Trained horses adapted to a high fat diet showed greater metabolic adjustment during exercise than trained horses adapted to a high starch and sugar diet, potentially allowing them to better meet energy demands. POTENTIAL RELEVANCE: Nutrition and exercise impact glucose and insulin dynamics, potentially influencing health and performance. PMID- 17402497 TI - Plasma glutamine concentrations in the horse following feeding and oral glutamine supplementation. AB - REASONS FOR PERFORMING STUDY: Pharmacological benefits of glutamine supplementation have been shown in athletically and clinically stressed human subjects. In the horse, infection and intense exercise have also been shown to significantly decrease plasma glutamine concentrations, but little is known on how best to supplement. OBJECTIVE: To evaluate whether ingestion of different foodstuffs, with or without L-glutamine (G) or a peptide (Pep) containing 31.5% w/w G in a water-stable form, could affect plasma glutamine concentrations (P GC). MATERIALS AND METHODS: Nine feeds (molassed sugar beet-pulp (mSB); naked oats (nO); commercial mix (CM); mSB with 30 or 60 mg/kg bwt G or the G-molar equivalent of Pep; and CM with 60 mg/kg bwt G or equivalent Pep) were offered to 6 healthy mature horses on different days following overnight food restriction. The changes in P-GC were monitored for 8 h post feeding. RESULTS: After 1.5 h mean +/- s.d. AP-GC were -0.9 +/- 10.2% (mSB), +12.5 +/- 7.1% (nO) and +44.7 +/- 15.9% (CM; P<0.05). deltaP-GC with mSB supplemented with G was +60.9 +/- 30.0% (30 mg; P<0.05) and +156.8 +/- 34.6% (60 mg; P<0.05) at 1 h; deltaP-GC with Pep was 51.0 +/- 31.0% (30 mg equivalent, P<0.05) and +91.1 +/- 9.5% (60 mg equivalent, P<0.05) at 1 h. After 10 days of supplementation with 60 mg/kg bwt G, AP-GC following a further 60 mg/kg bwt G challenge showed a similar increase at 1 h of +154.3 +/- 37.9%; prevalues were unchanged. G and Pep added to CM, increased P-GC by 246.3 +/- 55.3 (+99.2%) and 252.3 +/- 94.2 micromol/l (96.7%) at 1.5 h with concentrations still above prevalues at 8 h (P<0.05). Apart from the CM (with or without supplement), pre P-GC was always regained by 4 h. Plasma NH3 and plasma protein concentrations were unaffected by supplementation with G or Pep. CONCLUSION: P-GC may be modified by appropriate supplementation with no apparent adverse effects. POTENTIAL RELEVANCE: Increasing P-GC through appropriate supplementation may be of benefit in the athletically or clinically stressed horse with lowered plasma glutamine concentrations. PMID- 17402498 TI - Glycaemic and insulinaemic response of quarter horses to concentrates high in fat and low in soluble carbohydrates. AB - REASONS FOR PERFORMING STUDY: Quarter Horses are particularly susceptible to polysaccharide storage myopathy (PSSM). Nutritional therapy and possibly prophylaxis includes fat-supplemented diets whilst starch supply should be kept to a minimum. OBJECTIVES: To investigate the glycaemic and insulinaemic response of clinically normal Quarter Horses to concentrates high in fat and low in starch. METHODS: Twelve Quarter Horses were studied. The precondition for inclusion in the study population was that the horses had not shown clinical signs of myopathy. The Quarter Horses were fed according to a 4 x 4 Latin square design haylage plus isocaloric concentrates based on barley and oats as control (CO), sugar beet pulp, grass meal and soybean oil (SB), rice bran and grass meal (RB) and rice bran, grass meal, sugar beet pulp and soybean oil (CP), each over 2 weeks after 1 week of adaptation. At the end of each period, 1 kg of concentrate was fed and blood sampled 0, 30, 60, 90, 120, 180, 240, and 300 min post prandial (ppr.). Creatine kinase (CK; 0 min ppr. only), glucose and insulin were analysed. Glycaemic and insulinaemic index was calculated from each concentrates area under the curve (AUC) relative to CO. RESULTS: Rice bran containing concentrates were partially refused at the beginning of the trial periods. CK activity, and glucose and insulin patterns (ppr. mean, peak, AUC, index) were highest with CO (P<0.05). The correlation between glucose and insulin (P<0.001) ranged from r = 0.570 (CO) to r = 0.364 (RB). Basal CK was highly correlated (P<0.001) to the mean ppr. plasma glucose. CONCLUSION: Rice bran, sugar beet pulp, grass meal and soybean oil are suitable to include in concentrates that induce low glycaemic and insulinaemic response. Its acceptance seems to be a question of habituation. Although the mechanism is not completely understood, glucose patterns and basal CK activity are highly positively correlated. Both were affected by the type of concentrate used. POTENTIAL RELEVANCE: Results support the suggestion that low starch and high-fat feeding may be helpful not only in the nutrition of affected horses, but also to prevent future PSSM associated discomfort in particularly susceptible horses. PMID- 17402499 TI - Effects on exercise response, fluid and acid-base balance of protein intake from forage-only diets in standardbred horses. AB - REASON FOR PERFORMING STUDY: High-energy forage might be an alternative to concentrates for performance horses and such forage can be produced by an early cut. However, early cut forage is high in crude protein (CP), which may result in an excessive CP intake. OBJECTIVES: To investigate how CP intake affects nitrogen (N), fluid and acid-base balance, and exercise response in horses fed high-energy forage diets. The hypothesis was that high CP intake causes acidosis, and alters fluid balance and response to intensive exercise. METHODS: Two forage-only diets based on high-energy grass forage were fed for 23 days in a crossover design to 6 Standardbred horses in racing condition. One forage diet provided a high (HP) CP (16.6%) intake and the other diet provided recommended intake (RP) of CP (12.5%) for racehorses. The horses had intensive exercise twice and slow exercise 1-3 times every week. At the beginning and end of each period, faeces and urine were collected for 48 and 72 h, respectively and analysed for dry matter, pH and N content. At Days 19 and 23 in each period 2 race-like exercise tests were performed, a standardised treadmill test and a field test on a race track. Blood samples were taken before, during and after the tests and analysed for sodium (Na), potassium (K), chloride (CI), total plasma proteins (TPP), TCO2, urea, pH and lactate. The strong ion difference (SID) was calculated and heart rate and respiratory rate was also recorded. RESULTS: There was a decrease in urinary pH and an increase in N excretion, blood urea, water intake, urine volume and faecal water content on the HP diet. Total water intake was higher than the increase in urinary and faecal water loss indicating increased evaporative losses on the HP diet. During the exercise tests there were no significant differences between diets in TPP, plasma lactate, blood Na, K, Cl, TCO2, pH, SID and respiratory or heart rates. CONCLUSION: Feeding a forage-only diet with a CP intake corresponding to 160% of the requirement caused an increase in N excretion within 36-48 h after the diet was introduced and alterations in fluid and acid base balance at rest. POTENTIAL RELEVANCE: The increased urine and probably also evaporative fluid losses suggest that feeding a HP diet will cause an unnecessary challenge for horses during prolonged exercise. PMID- 17402500 TI - The effect of milk basic protein supplementation on bone metabolism during training of young thoroughbred racehorses. AB - REASONS FOR PERFORMING STUDY: In laboratory animals, man and cell culture experiments, milk basic protein was reported to suppress bone resorption and promote bone formation. However, no studies in horses have previously examined the effect of milk basic protein. OBJECTIVES: To evaluate the effect of milk basic protein supplementation on bone metabolism in young Thoroughbred horses in training. METHODS: Twenty 2-year-old horses in training were used for 90 days in this study. The treatment group was fed a basal diet with 1 g of milk basic protein and the control group a basal diet only. Blood samples were collected on Days 0, 45 and 90 to determine serum calcium (Ca) and biochemical markers of bone metabolism. Radiographs were taken at the start and end of the study to determine radiographic bone aluminium equivalence (RBAE). RESULTS: Serum osteocalcin (OC) was significantly higher at Day 45 after the beginning of the study in the treatment group compared to that in the control group. The treatment group showed a greater increase in the total RBAE change at the end of this study compared to that in the control group. However, there were no significant differences in serum Ca and carboxy-terminal telopeptide of type I collagen (ICTP) between groups. CONCLUSIONS AND POTENTIAL RELEVANCE: These findings provide preliminary evidence that milk basic protein has an effect on bone formation in 2-year-old Thoroughbred horses in training. However, further studies in larger groups of horses are now required to substantiate our findings. PMID- 17402501 TI - Effects of different levels of calcium and phosphorus intake on calcium homeostasis in exercising horses. AB - HYPOTHESIS: Increasing levels of dietary calcium (Ca) and phosphorous (P) might have a negative impact on parathyroid hormone (PTH) response and result in a more pronounced hypocalcaemia during high-speed exercise in horses. MATERIAL AND METHODS: In successive order, five trained horses were fed 33 g Ca and 19 g P (approximately 100% NRC, adequate intake), 64 g Ca and 38 g P (moderately high intake) or 96 g Ca and 56 g P (high intake). Each horse was adapted to each diet over a 21 day period before undergoing a standardised exercise test (SET) on a treadmill. The SET comprised 5 steps (each step 4 min duration, 3% incline, first step 5 m/sec, followed by increments of 1 m/sec). Blood samples were taken at defined times. RESULTS: Blood lactate, plasma intact PTH (PTHi) and plasma inorganic P (Pi) increased during exercise (P<0.05), blood pH and blood ionised Ca2+ (Ca2+, adjusted to a pH of 7.4) declined during SETs (P<0.05) whereas total plasma Ca (CaT) and Mg (MgT) remained constant. The most exaggerated drop in Ca2+ and lactate peaks were found for the adequate Ca and P intake at 7, 8, and 9 m/sec during SET (treatment P<0.05). At 120 min after exercise, Ca2+ levels exceeded resting concentrations with higher Ca2+ values for the higher Ca intake (Ca2+: adequate 1.58 +/- 0.07 mmol/, moderately high 1.63 +/- 0.04 mmol/l, high 1.65 +/- 0.02 mmol/l, P<0.05). Concomitantly, 120 min after exercise, PTHi returned to basal concentrations with higher PTH concentrations for the adequate Ca intake than the other two treatments (PTHi: adequate 45.6 +/- 72.8 pg/ml, moderately high 11.6 +/- 13.6 pg/ml, high 2.6 +/- 2.3 pg/ml, P<0.05). During recovery (30 min, 120 min and 24 h after exercise) MgT concentrations were depressed (P<0.05) for the high Ca intake (P<0.05). CONCLUSION: These results suggest that high dietary Ca and P intake did not impair PTH responses during exercise and recovery, but might depress Mg metabolism. POTENTIAL RELEVANCE: High Ca and P intake might have some potential to increase the availability of blood Ca2+ and Pi in the recovery phase after intensive exercise, but detrimental effects on Mg homeostasis might offset this potential benefit. PMID- 17402502 TI - Urinary excretion of dietary contaminants in horses. AB - REASONS FOR PERFORMING STUDY: Presence of drugs is completely prohibited in post racing urine samples by most of racing and competition authorities, even if environmental contamination might occur. OBJECTIVES: To assess the daily dose of several contaminants absorbed through the diet that would result in detectable concentrations in urine. METHODS: Caffeine, theobromine, theophylline, atropine, scopolamine, bufotenine, DMT or morphine were administered orally to 6 horses, in different dosages, for 3 days before their urine was sampled for regular anti doping tests. RESULTS: Theobromine, theophylline, bufotenine and morphine daily intake >10 mg, 2 mg, 10 mg and 200 microg, respectively, by a performance horse, were found to result in detectable urinary concentrations. At the 2 tested doses, atropine (5 and 15 mg) and dimethyltryptamine (3 and 10 mg) were not detected in urine. For caffeine and scopolamine, even the lowest dosage tested (5 mg/horse/day and 2 mg/horse/day respectively) induced detectable concentrations of the molecule in urine. CONCLUSIONS: Horses fed dietary contaminants, even at level much below the effective dosage, may be positive to antidoping urine analysis. Further research is needed to gain more confident results on a daily safe intake for caffeine and scopolamine. POTENTIAL RELEVANCE: Selection of feed materials appears to be of great importance to prevent non voluntary positive result to anti-doping tests. PMID- 17402503 TI - Effect of omeprazole on markers of performance in gastric ulcer-free standardbred horses. AB - REASONS FOR PERFORMING STUDY: A large percentage of performance horses develop gastric ulcers and many of those horses are treated with omeprazole. Unfortunately, no data have been published on the effects of the drug on markers of performance in animals without ulcers. HYPOTHESIS: Omeprazole would alter markers of aerobic and anaerobic performance. METHODS: Ten unfit, healthy, ulcer free, Standardbred mares were administered either control (CON; oral apple sauce, 20 ml) or omeprazole (OP; oral paste, 4 mg/kg bwt s.i.d.) in a random crossover fashion with the investigators blind to the treatment. Treatments were administered for 7 days prior to performing an incremental exercise test (GXT) on a high-speed treadmill. Endoscopic examinations were performed just prior to the trial to verify that the mares were ulcer-free. During the GXT, the mares ran on a treadmill up a 6% grade to measure maximal oxygen consumption (VO2max), run time (RT), velocity at VO2max, maximal velocity (Vmax), packed cell volume (PCV), plasma lactate concentration (LA) and plasma protein concentration (TP). Measurements were recorded at rest, at the end of each 1 min step of the GXT and at 2 and 5 min post GXT. Data were analysed using ANOVA for repeated measures and t tests for paired comparisons. RESULTS: There was no effect (P>0.05) of omeprazole on VO2max; velocity at VO2max; RT; Vmax; 2 min recovery plasma LA. Nor were there any changes (P>0.05) in the relationship between treadmill speed and VO2, PCV, TP, or plasma LA. CONCLUSIONS: Omeprazole does not appear to improve physiological markers of performance in healthy, ulcer free horses. POTENTIAL RELEVANCE: These data may benefit various authorities responsible for deciding administration and timing policies of omeprazole as well as clinicians and horse owners. PMID- 17402504 TI - Analytic and rule-based decision support tool for VDT workstation adjustment and computer accessories arrangement. AB - One important factor that leads to the development of musculoskeletal disorders (MSD) and cumulative trauma disorders (CTD) among visual display terminal (VDT) users is their work posture. While operating a VDT, a user's body posture is strongly influenced by the task, VDT workstation settings, and layout of computer accessories. This paper presents an analytic and rule-based decision support tool called EQ-DeX (an ergonomics and quantitative design expert system) that is developed to provide valid and practical recommendations regarding the adjustment of a VDT workstation and the arrangement of computer accessories. The paper explains the structure and components of EQ-DeX, input data, rules, and adjustment and arrangement algorithms. From input information such as gender, age, body height, task, etc., EQ-DeX uses analytic and rule-based algorithms to estimate quantitative settings of a computer table and a chair, as well as locations of computer accessories such as monitor, document holder, keyboard, and mouse. With the input and output screens that are designed using the concept of usability, the interactions between the user and EQ-DeX are convenient. Examples are also presented to demonstrate the recommendations generated by EQ-DeX. PMID- 17402505 TI - The relationship between job satisfaction and job factors in industrial work design: a case study of the automotive industries in Malaysia. AB - A survey was conducted to investigate the relationship between job satisfaction and job factors that affect work design in two automotives manufacturing companies in Malaysia. A set of multiple choices questionnaires was developed and data were collected by interviewing the employees at the production plant. Hundred and seventy male subjects between the ages of 18 to 40 years with the mean age of 26.8 and SD of 5.3 years and mean work experience of 6.5 and SD of 4.9 years took part in the survey. The survey focused on job factors, i.e. skill variety, task identity, task significance, autonomy and feedback. The results support the previous findings that job factors are significantly correlated to job satisfaction. Furthermore, it also highlights the significant influence of age, work experience and marital status. PMID- 17402506 TI - Effects of VDT monitor placement and single versus bifocal glasses on somatic discomfort and postural profiles in data entry tasks. AB - A study was conducted to determine the effects of VDT monitor positions and the use of single vision versus bifocal glasses on somatic elements in the data entry task. Eight male subjects performed data entry using a word processor in eight half-hour sessions with the four different monitor placements, i.e. "eye-level", "shoulder-level-front", "shoulder-level-side", and "sunken-level", wearing the two types of glasses. A subjective discomfort rating questionnaire covering 12 somatic elements was completed by the subject after each session. The head inclination and angle of gaze to the monitor were measured with a goniometer. The results revealed that the somatic elements which were affected significantly by the placement of the VDT monitor and the type of glasses were discomfort in the neck and back regions and eyestrain, respectively. The neck-back discomfort scores were highest at the "eye-level", lowest at the "sunken-level", and intermediate at the "shoulder-level-side" position. The "shoulder-level-front" position was not significantly different in the discomfort from other three positions. The eyestrain was significantly greater with the bifocal than with the single vision glasses. The lower the monitor was placed, the more forward was the head and gaze inclined. The head was inclined less forward, or even more backward, and the gaze was inclined more forward, with the bifocal than with the single vision glasses. As a conclusion, the VDT operators were advised to avoid the "eye-level" and "shoulder-level-side" positions and to prefer the "sunken level" and "shoulder-level-front" positions as the first and second best choices, respectively. The preference becomes more critical for the wearers of bifocal glasses that suffer from postural constraints in viewing. PMID- 17402507 TI - Analysis of "Teno-uchi" maneuver in releasing Japanese-style bows based on muscle activities and forces acting on the bow. AB - Since a Japanese-style bow has a very complicated shape and structure, an archer has to apply the "Teno-uchi" maneuver including horizontally twisting torque, or "Nejiri", and sagittally down-pushing torque, or "Uwa-oshi", to the restoring bow in order to hit the target. The purpose of this study was to investigate the biomechanical relationship between the muscular activities of the left forearm and the operation of "Teno-uchi" maneuver. Surface EMG of left forearm muscles and the two kinds of torque acting on the bow around the time of release were recorded in 10 experienced subjects during arrow shooting. The "Biku", an involuntary resignation from release happening in the shooting, was also examined. Close analyses of the results revealed that activation of the extensor carpi ulnaris and extensor digitorum muscles together with inhibition of the flexor carpi ulnaris muscle brought about "Nejiri", while activation of the extensor carpi ulnaris as well as flexor carpi ulnaris muscles and inhibition of the extensor carpi redialis longus and extensor digitorum muscles gave rise to "Uwa-oshi", thus causing activities of trade-off nature in the extensor digitorum and flexor carpi ulnaris muscles for the "Nejiri" and "Uwa-oshi. The trade-off activities were presumably actualized through time-sharing coordination between the muscles. PMID- 17402508 TI - The effects of the load mass and load position on body sway in supporting a load on the back. AB - We examined the effects of a load's mass and position on body sway during standing with a load on the back. Three healthy male subjects participated in this experiment. The subjects supported loads of 23kg, 33kg, and 43kg on their backs using a carrier frame. They were asked to stand for 75s on a force platform with their eyes open while being as quiet as possible. Time series data of center of-pressure (COP) were collected at a sampling rate of 50Hz during the last 60s of the 75s standing period. The COP was measured under three conditions in terms of the load position on the frame: lower (close to the hip), middle, and upper (close to the shoulder). All subjects showed that the lower the position of the load, the more anteriorly the mean COP coordinate was located in the anteroposterior (AP) direction, and the smaller the total distance of the COP trajectories became. Regarding carrying the heavier loads, each subject showed a specific tendency in the mean AP coordinate. The three subjects had different physical characteristics in terms of body height and experience at carrying heavy loads. These results suggest that the examintion of the COP in a static posture can help our understanding of individual information on the posture supporting loads and the general positioning of the body. PMID- 17402509 TI - Effects of barefoot habituation in winter on thermal and hormonal responses in young children--a preliminary study. AB - This study investigated how socks-wearing habit or habitual barefoot in the cold winter affected skin temperatures of distal lower extremities, the urinary excretion of adrenaline, noradrenaline and cortsisol in young children. In Experiment I for preschool children, the measurements of foot and leg skin temperatures were conducted for 30 min in the classroom controlled at 23+/-2 degres C and 50+/-5%RH, and the excretion of urinary catecholamines and cortisol during nocturnal sleep were analyzed. In Experiment II for elementary school children, nocturnal secretion of urinary catecholamines and cortisol was analyzed. While leg skin temperature tended to be lower in barefoot group than in socks group during Experiment I, foot skin temperature was not significantly different between the two groups. Fall of leg skin temperature during 30 min measurement tended to be smaller in barefoot group than in socks group. Urine volume and urinary excretion of cortisol tended to be greater in barefoot group than in socks group for preschool children. Urinary noradrenaline was significantly greater and cortisol tended to be greater in barefoot group than in socks group for elementary school children. Considering that most of the findings shown above were in the proximity of the established level of statistical significance, it was provisionally concluded that young children with barefoot habituation might show more effective cold adaptation of metabolic type than those without the habituation do, by keeping their skin temperatures higher even in the cold and enhancing the metabolic rate. PMID- 17402510 TI - Assessment of diaphragmatic levels on plain chest radiograph--a new approach. AB - BACKGROUND: The aim of this study was to provide an alternate simple linear measurement of diaphragmatic levels from a relatively fixed position, the lung apex, on chest radiographs. STUDY DESIGN: 526 consecutive posteroanterior (PA) chest radiographs of Nigerian adult subjects reported as normal were evaluated. The 'Lung apex to diaphragmatic apex' (LATDA) distances in centimeters for both hemidiaphragms were obtained from the chest radiographs. RESULTS: There were 210 males and 316 females ranging in age from 15 to 85 years. The mean ranges of LATDA values on the right were 21.2 cm to 23.8 cm and 20.8 cm to 22.7 cm, with total means of 22.9 cm and 21.5 cm in males and females respectively. On the left, the ranges were 23.1cm to 25.5 cm and 21.6 cm to 23.5 cm with total means of 24.4 cm and 22.6 cm in males and females respectively. Left LATDA was higher in 95% of subjects and higher LATDA values represent lower hemidiaphragms while low LATDA values indicate elevated hemidiaphragms. LATDA measurements were statistically higher in males for all age groups. Mean differences between right and left LATDA were 1.5 cm and 1.2 cm in males and females respectively. CONCLUSION: From this study, the lower limits of normal LATDA values in females and males are 16 cm and 18 cm while upper limits are 28 cm and 30 cm respectively. A "rule of twos" that could be applied to routine use in clinical practice is suggested thus: Left LATDA value is 2 cm higher than right LATDA value and LATDA values in males are 2 cm higher than in females. PMID- 17402511 TI - Antibiotics prophylaxis for digital-guided transrectal tru-cut needle biopsy of the prostate. AB - BACKGROUND: Prostate biopsy is essential to diagnose prostate cancer. The complications need to be kept in view. AIM: To explore the outcome of prostate biopsy using Gentamicin 280 mg by intramuscular injection as prophylaxis against infection. PATIENTS AND METHODS: Patients aged over 40 years seen in Sophia Clinic Port Harcourt over a 5-year period with clinical suspicion of prostate cancer had transrectal biopsy of the prostate after written consent. Each received Gentamicin 280 mg intramuscularly one hour before biopsy without anaesthesia and without rectal washout. Biopsy was done with an 18 F Tru-cut biopsy needle guided digitally. The histology reports were assessed. Biopsy was repeated at different sessions if earlier reports were inconsistent with clinical suspicion. The patients were reviewed one week after biopsy. RESULTS: One hundred and nine subjects aged 48-95 years had 111 prostate core needle biopsies. Biopsies positive for prostatic disease were 108 (97.3%) including adenocarcinoma 98 (88.3%) and benign prostatic hyperplasia (BPH) 10 (9.0%). Three specimens (2.7%) showed no prostate tissue. In 2 specimens from a patient with osteoblastic radiological features of prostate cancer, the histology report indicated BPH. On seven occasions (63%), patients had severe pain during the procedure but did not require analgesia. Transient rectal bleeding occurred in 15 patients soon after the procedure. Complications noted within a week after the procedure included fever in four patients, urinary tract infection in four, haematuria in eight, septicaemia in one and a horse-shoe perianal abscess in one. CONCLUSION: Serious complications after transrectal Tru-cut biopsy of the prostate are uncommon. Septic complications may have been reduced by the antimicrobial prophylaxis. PMID- 17402512 TI - A five-year review of upper airway obstruction in the immediate general anaesthetic post-operative period in Enugu, Nigeria, 1990-1994. AB - AIM: To determine the incidence and causes of upper airway obstruction in the Post Anaesthesia Recovery Room (PARR) of the University of Nigeria Teaching Hospital, Enugu, from January 1990 to December 1994 MATERIALS AND METHODS: A retrospective study of 4500 anaesthetic charts from January 1990 to December 1994 was undertaken. Cases where upper respiratory tract obstruction (URTO) occurred were retrieved and reviewed. Information sought for included patient's age, sex, pre-operative physical status (ASA), type of surgery, probable cause of URTO, and management outcome. RESULTS: Twenty-five patients had URTO during the study period giving an overall incidence of 0.56% or 56 in 10,000 general anaesthetics. The most common cause of URTO in the recovery room was laryngospasm followed by the backward fall of patients tongue. CONCLUSION: This study has shown that upper airway obstruction is a fairly common complication in the post-anesthesia recovery room and that laryngospasm is the most important aetiology. PMID- 17402513 TI - The most appropriate site for endoscopic biopsy for the detection of H. pylori among Nigerians in Ibadan. AB - BACKGROUND: Helicobacter pylori is a significant aetiological factor for acid peptic diseases and gastric cancer. Detection of the organism in gastric mucosal biopsies is important, hence the need to ascertain the optimal site for biopsy that will facilitate identification of the organism. STUDY DESIGN: The study was carried out by obtaining directed gastric and duodenal endoscopic biopsies from twenty-five adult Nigerian patients clinically diagnosed to have gastroduodenitis at the University College Hospital, Ibadan, Nigeria. Biopsies were obtained from the gastric body, pyloric antrum, first and second parts of the duodenum at endoscopy. The biopsy specimens were tested for H. pylori by Campylobacter-like organism (CLO) test and histology. RESULTS: Positive results for H. pylori by CLO test were detected in 67% each for the biopsies taken from the gastric antrum and incisura angularis; and 28% and 17% for those taken from first and second parts of the duodenum respectively. There was no benefit in taking additional biopsy from incisura angularis to that from the antrum. Helicobacter pylori was better detected in the mucosa of the antrum (72%) than that of the duodenum (28%), p < 0.05. The organism was detected in 28% by histological examination of the tissue specimen of the patients compared to 72% by CLO test, p < 0.05. CONCLUSION: The study shows that the detection of H. pylori by invasive technique is better obtained by taking biopsy at the gastric antrum in Nigerian patients with gastroduodenitis. Furthermore, the CLO test yields more positive results than histological evaluation in the detection of the infection. PMID- 17402514 TI - Causes of limb amputations in Nigerian children. AB - BACKGROUND: Most reports on amputations in Nigeria have been on adults. Few published data exist on the causes of amputations in children. METHODS: A retrospective analysis of all children who had limb amputations from January 1998 to December 2004 at the Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife was made. RESULTS: Thirty five (32.7%) out of 107 patients who had major limb amputations were children. The mean age was 9.9 years. Trauma accounted for 74.3% of the amputations, 92.3% of which had developed gangrene from treatment of simple fractures by traditional bone setters. Fifty one point four percent of the amputations were in the upper limbs. The most common complications were anaemia (88.6%) and wound infection (68.6%). One patient had tetanus while one died from septicaemia. Only two patients had prosthetic fitting following amputation. CONCLUSION: Most amputations in Nigerian children are due to traditional bone setters who manage simple, straight forward fractures in children causing gangrene of the limbs. PMID- 17402515 TI - Malignant skin tumor in Blacks: experience in a teaching hospital. AB - BACKGROUND: Basal Cell Carcinoma (BCC) is said to be the most common human skin malignancy. Its common presentation is in a Caucasian male older than 60 years, and patients could present with multiple co-existing tumors. METHODS: We retrospectively review 146 patients treated for skin tumors in our tertiary hospital from January 1979 to December 2002. Cases notes and histopathology report cards of the patients were the source where information on individual patients was extracted. Patients treated for cutaneous melanoma were excluded from the study. RESULTS: This study showed that Squamous Cell Carcinoma (SCC) is the most common non melanomatous skin tumor in our environment. The study revealed only 8 (5.5%) cases of basal cell carcinoma which is in contrast to what is obtained in the Europe and North America. CONCLUSION: Unlike in the Caucasians, squamous cell carcinoma is the most common non-melanomatous skin tumor in our environment. Most of our cases resulted from chronic leg ulcers secondary to poorly treated traumatic ulcers while others are due to chronic burn scar. The tumor is usually more aggressive and more prone to metastasis resulting in severe morbidity and sometimes loss of limb in the affected patient. PMID- 17402516 TI - Microalbuminuria and clinical correlates in black African patients with type 2 diabetes. AB - BACKGROUND: Diabetes mellitus is a major cause of morbidity and mortality as a result of its complications. Long-term complications of diabetes mellitus have been linked to poor glycemic control. Diabetic nephropathy is the leading cause of end stage renal disease in the developed world and third leading cause in Nigeria. Various independent risk factors have been identified as predictors of diabetic nephropathy. One of such factors is urinary albumin excretion. OBJECTIVE: This study set out to determine the prevalence of microalbuminuria (MA), glycemic control and the relationship between urinary albumin excretion (UAE) and other known predictors of diabetic nephropathy in type 2 diabetic patients with disease duration greater than 5 years. STUDY DESIGN: Fifty non proteinuric patients were selected consecutively for this cross-sectional study. Urinary albumin concentration was determined in a timed overnight urine sample by immunoturbidimetry and glycated haemoglobin was determined using boronate affinity method. Comparison was made between the patients with microalbuminuria and those with normal albumin excretion and the most likely predictors of urinary albumin excretion were determined. RESULTS: Fifty-two percent of the patients studied had good glycaemic control (HbA1c < 7%) while 83% had microalbuminuria (UAE 20-200 mg/min). There was no significant correlation between the UAE and the HbA1c. UAE however correlated significantly with the age and diastolic blood pressure (DBP) of the patients. CONCLUSION: Almost half of the patient population had suboptimal glycemic control. There was a high prevalence of microalbuminuria amongst the patients studied. Significant predictors of UAE in this study were the age of the patients and DBP but not HbA1c. PMID- 17402517 TI - Types of crises in sickle cell disease patients presenting at the haematology day care unit (HDCU), University College Hospital (UCH), Ibadan. AB - OBJECTIVE: The aim of the study was to determine the types of crises in Sickle Cell Disease (SCD) patients seen at the Haematology Day Care Unit (HDCU) of the University College Hospital (UCH), Ibadan. MATERIAL AND METHODS: The study is a retrospective survey using the registers of all patients seen in the year 2001 (January-December). Data extracted included demographic parameters, types of crises, haematocrit level and outcome of treatment received. The patients studied were both HbSS and HbSC in crises. RESULT: There were five hundred and eight HbSS and thirty seven HbSC patients seen during the survey period. Vaso-occlusive crisis had the highest prevalence of 91.6%, while haemolytic crisis had the least (0.5%). More females (56.7%) had crises than males (43.3%) in the vaso-occlusive group. The relationship between crises type and sex was not significant (p = 0.282). Young adults (20-29 years) had more crises (49.9%) than other age groups but crises were rare in children (0.2%). There was, however, a significant relationship between age group and crises type (p < 0.00). Eighty one percent of the subjects had PCV > 20%. The type of crisis is related significantly to the treatment received (p < 0.000). Fifty six percent of the subjects required analgesics alone while was admitted for blood transfusion and other medical, surgical, gynaecological and obstetric complications. There was only 1 (0.1%) recorded death within the 12 month survey period in a 31 year-old HbSC female who had painful and haemolytic crisis. CONCLUSION: There were six types of crises seen in the sickle cell disease subjects namely vaso-occlusive, sequestration, infarctive, aplastic, haemolytic and bone pain crises. Vaso-occlusive crisis was the most common and haemolytic crises the least. Vaso-occlusive crisis was more common among young adults (20-29 year age group). The mortality rate was found to be 0.1%. PMID- 17402518 TI - An unusual complication of the use of stapler after Hartmann's procedure. AB - BACKGROUND: Circular anastomotic staplers have been used in colorectal surgery for several years with low complication rates. We report here an extremely rare case of direct colovaginal anastomosis and rectovaginal fistula occurred by using a 31 end-to-end anastomosis stapler after reversal of Hartmann's Procedure. STUDY DESIGN: A 33-year-old woman with complaint of rectal bleeding had undergone low anterior resection and end colostomy for rectal adenocarcinoma in 1995. In spite of making recommendation for colostomy closure, she had postponed the operation until 2003 due to afraid of incontinence. After reversal of Hartmann's procedure achieved by using circular stapler and diverting ileostomy, she developed complaints of vaginal gas and minimal vaginal fecal discharge. RESULT: Endoscopic examination and imaging methods revealed a rectovaginal fistula with a blind Hartmann's pouch. After a certain verification of complexity of the case, a reoperation which comprises true intestinal anastomosis and repair of fistula was performed. Postoperative period was uneventful except an incisional infection, and ileostomy was closed subsequently. CONCLUSION: A rectovaginal fistula, though encountered rarely, must be taken into account in cases with suspicious complaints after using circular stapler for pelvic intestinal anastomosis. Careful placement of stapler in accurate direction has a great importance to avoid stapling related complications. PMID- 17402519 TI - Anisometropia in a tropical hospital. AB - BACKGROUND: This was to determine the prevalence and distribution of anisometropia and to study the difference in refraction between the eyes of patients with anisometropia and look for evidence of laterality, if any. STUDY DESIGN: A prospective study of 1819 consecutive new patients seen in our clinic with no other pathology except refractive error between November 1998 and December 2002 were included in the study. Information obtained included age, sex, visual acuity, type and degree of anisometropic error. Non-parametric statistical methods were used for analysis. RESULTS: Of 1819 patients seen, 815 (44.81%) had anisometropia. There were 389 males and 426 females with a male to female ratio of 1.00 to 1.1 and a mean age of 35.15 (standard deviation 17.38). Of the 815 patients, 476 (58.40%) had spherical anisometropia and 339 (41.60%) had cylindrical anisometropia. Myopic anisometropia (253 (31.04%) was the most common. Of the spherical anisometropic errors excluding the mixed types, majority (245 (57.65%) had a difference of 0.25 dioptres and 28 (6.59%) had a difference of 2.00 dioptres and above. The highest difference of 8 dioptres was found in only one patient. The mean error degrees were higher on the left in both myopia and hypermetropia. CONCLUSION: Anisometropia is quite common in this environment. There is a tendency for the left to be higher contrary to previous findings. In order to allow for data comparison between countries and give room for the occasional high difference between eyes, it may be better to use both eyes in data analysis contrary to previous impression. PMID- 17402520 TI - An audit of gunshot injuries seen in the accident and emergency department of a Nigerian tertiary hospital. AB - AIM: An audit of gunshot injuries was conducted at the University College Hospital to determine the pattern of injuries. MATERIALS AND METHODS: All records of patients who presented with gunshot injuries to the accident and emergency department of the University College Hospital for one year were retrieved. Data obtained from the records included demographic data (age and sex), anatomical regions involved, the type of missile classification, the circumstances surrounding the incident and the patient's vital signs at presentation. The severity of injury was computed using the injury severity score. RESULTS: Eighty out of ninety patients' records were seen. There were 15 females (18.75%) and 65 (81.25%) males with mean ages of 25.60 years and 35.42 years respectively . The most common cause of gunshot injuries was armed robbery attack (71.3%) while the extremities (upper limbs and lower limbs) were the most common areas of injury (61.3%). Armed robbery was the most frequent cause of injuries. CONCLUSION: Low velocity missiles are predominant. The lower limb is the most commonly injured area. Armed robbery is the most common cause of such injuries. Most gunshot injuries are physiologically stable at presentation. Suicidal intent is still rare. PMID- 17402521 TI - Prostate cancer in Maiduguri. AB - BACKGROUND: Prostate cancer is the sixth most important cancer in the world, and its incidence in blacks has been on the increase. It is a very important cause of morbidity and mortality. In North-eastern Nigeria, reports on prostate cancer have been scarce. The aim of this paper is to highlight the clinico-pathological features and management of prostate cancer in north-eastern Nigeria. STUDY DESIGN: The clinical records and histopathological slides of all prostate cancer patients seen at the UMTH from January 1987 to December 2004 were retrospectively reviewed. The total number of cancers diagnosed during this period was also obtained from the cancer registry. RESULTS: Prostate cancer was seen in 165 patients and accounted for 6.15% of cancers diagnosed in this hospital. Majority of patients (56%) were aged 65 years and below and most presenting symptoms were similar to those of benign prostatic hypertrophy (BPH). The diagnosis was made in most cases on the basis of DRE, USS, PSA, and confirmed by trucut prostatic biopsy. Since most cases were advanced, the most frequent palliative treatment was subcapsular orchidectomy, which was psychologically satisfying to patients. Follow-up was very poor, only 26% at 3 years. CONCLUSION: Since most patients present late we conclude that prostate cancer should be suspected in men of 50 years and above who present with symptoms of prostatism and should be investigated promptly. Furthermore, aggressive screening of men in this age group would facilitate early diagnosis. PMID- 17402522 TI - Informed consent for surgery: a historical review. AB - BACKGROUND: Many surgeons see the process of obtaining informed consent for an operation from a patient beginning and ending with the signature of the patient on the consent form. Indeed this is not so as this process is deeper than that and all operating specialists need to understand the origin of this process, the developments and the areas where interpretations may be difficult. This is to ensure that implementation of the process of informed consent is mutually beneficient to both patient and doctor. METHOD: Literature dealing with the origin and history of informed consent, its principles, various interpretations and experiences in various subspecialties was reviewed. RESULTS: The informed consent process arose from the proceedings of the Nuremberg trial in 1947 to ensure that such atrocities that were committed on human beings in the pursuit of medical research were never repeated. The basic tenet of the informed consent rests on the autonomy of the patient which is explained as being the legal embodiment of the concept that each individual has the right to make decisions affecting his/her well-being. Areas of possible controversy which includes the interpretation of competence and understanding of the patient in the light of adequate information from the doctor were also highlighted. CONCLUSION: It is apparent from the wealth of literature on this subject that there is still a problem regarding the lack of guidelines towards achieving the perfect informed consent. PMID- 17402523 TI - Surgical management of advanced hypopharyngeal carcinoma: a case report and literature review. AB - BACKGROUND: Surgical resection of advanced primary head and neck cancers especially of the upper aerodigestive tract like hypopharyngeal cancer, usually leave a very big defect, which has to be reconstructed in order to maintain the gastropharyngeal continuity. This oncological surgery is best performed at the same sitting as a one staged procedure. STUDY DESIGN: A clinical case of a 56 year-old female with advanced hypopharyngeal cancer who had a single stage total pharyngolaryngectomy and partial oesophagectomy with reconstruction of the upper aerodigestive tract using pectoralis major pedicled flap and post operative adjunct chemo-radiation therapy. RESULT: Functionally, our patient had good swallowing reflex, fed on normal diet free of feeding tube and no gastric reflux. There was clinical evidence of weight gain. No evidence of flap complications post surgery. CONCLUSION: The rarity of this procedure and its advantages in the management of an advanced hypopharyngeal cancer in this environment are discussed. PMID- 17402524 TI - Kocher-Debre-Semelaigne syndrome: a case report. AB - BACKGROUND: Kocher-Debre-Semelaigne (KDS) syndrome is a myopathy of hypothyroidism associated with pseudohypertrophy in infancy or childhood. There are few reported cases of KDS syndrome in the literature. STUDY DESIGN: We present a 5-year-old boy with poor growth and delayed dental and motor development. There was no family history. On examination he had coarse facies, large protruding tongue, athletic build, short stature and mental retardation. The diagnosis of KDS syndrome was based on laboratory and radiologic evidence of congenital hypothyroidism and muscle enlargement. He was started on L-thyroxine at the dose of 4ug/kg/day, and he has shown marked increase in alertness with regressing muscle bulk after 4 weeks of treatment. A short review of the literature is also presented. PMID- 17402525 TI - Myelomatosis: a case report of an unusual presentation. AB - BACKGROUND: Multiple myeloma (MM) is a clonalexpansion of plasma cell characterized by skeletal dissemination of malignant plasma cells resulting in the production of homogenous monoclonal immunoglobulin termed the monoclonal (M) protein. MM is the most prevalent cancer after non-Hodgkin's lymphoma and is responsible for 2% of all cancer deaths. Several unusual presentation of MM have been described in the literature. Many serial reports have documented progression of Plasmacytoma to MM, or relapsed into MM after radiation therapy. STUDY DESIGN: This is a case report of a 36 yr old man with a huge fungating right clavicular mass, with a histological of plasmacytoma (anaplastic variant). RESULT: The patient's tumour was resistant to radiotherapy, terminating as multiple myeloma, with rapid progression to death within 19 months post diagnosis. CONCLUSION: This report helps to validate the rapid progression of the anaplastic plasmacytoma variant to disseminated myeloma, and indicates the poor response to radiation therapy. Our patient showed the important poor prognostic features in some plasmacytomas that are characterized by rapid progression to MM, namely tumour size > 5 cm, young age and spinal cord compression. PMID- 17402526 TI - She knows what's coming. PMID- 17402527 TI - Agreeing across the ages. PMID- 17402528 TI - Recipe for change? PMID- 17402529 TI - Empowering older people. PMID- 17402530 TI - Falling targets. PMID- 17402531 TI - A Stirling centre. PMID- 17402532 TI - Finding professional fulfilment in caring for older people. AB - This paper describes the findings of a phenomenological study of professional fulfilment in nurses who care for older people. The author sought to uncover what brings career satisfaction to nurses who care for older people and subsequently what motivates these caregivers to continue to care. The findings have implications for clinical nurses who may see reflections of their own approaches to care in the narratives presented. The examples of care reported may inspire nurses who work with older adults by affirming their own nursing interventions. PMID- 17402533 TI - Nursing students and elder abuse: developing a learning resource. AB - This article describes the development of a learning resource designed to increase awareness of elder abuse among nursing students. The learning resource combines exercises, lectures, group work and the use of film. It encourages students to reflect on ways of identifying and preventing elder abuse in the increasingly wide range of settings where they are likely to work with older people. PMID- 17402534 TI - Medication issues for nursing older people (part 2). AB - This article aims to raise awareness about professional responsibility and accountability in medication issues that impact on older people. Nurses must recognise their responsibilities in regard to the safe administration of medicines and challenge poor drug administration practices. PMID- 17402535 TI - Patricia Cohen. PMID- 17402536 TI - [The skin as an immune organ: influence of therapeutic physical factors]. PMID- 17402537 TI - [Cardiovascular response in mountain-skiers on vacation at a mountain climatic health resort]. PMID- 17402538 TI - [Reparative effect of siliceous baths and their combination with laser and magnet therapies for trophic ulcers of venous etiology]. PMID- 17402539 TI - [Impact of naphthalan therapy on the clinical and functional status of patients with chronic obstructive lung disease]. PMID- 17402540 TI - [Non-drug therapy for metabolic disturbances in abdominal obesity]. PMID- 17402541 TI - [Complex use of physical factors to correct edematous fibrosclerotic panniculopathy]. PMID- 17402542 TI - [Chronopathology and chronotherapy for chronic pancreatitis (Part 1. "Seasonal rhythms and preventive therapy")]. PMID- 17402543 TI - [Intestinal dysbiosis and methods for its correction in patients with chronic salpingoophoritis]. PMID- 17402544 TI - [Therapeutic exercises in children with noninfectious craniocervical transition lesions]. PMID- 17402545 TI - [Lidase electrophoresis and the immunomodulator polyoxidonium in the rehabilitative treatment of patients with Peyronie's disease]. PMID- 17402546 TI - [Evaluation of the efficiency of sanatorium-and-spa treatment]. PMID- 17402547 TI - [Impact of thalassotherapy on healthy individuals' microcirculation]. PMID- 17402548 TI - [Physical rehabilitation of patients with neurological manifestations of cervical osteochondrosis at a resort]. PMID- 17402549 TI - [Mineral waters of the European part of Russia]. PMID- 17402550 TI - [Complex rehabilitative treatment for seronegative spondyloarthropathy. A manual for the physician]. PMID- 17402551 TI - Genetic polymorphism of cytochrome P450 and methods for its determination. AB - The majority of human P450 dependent drug metabolism is carried out by polymorphic enzymes which can alter plasma concentration of the pharmacological active substance followed by an enhanced or suppressed pharmacological effect. The response of individual patients to drugs can be affected by variations in DNA sequence mainly by single nucleotide polymorphisms (SNPs). Knowledge of functionally important SNPs prior to the drug administrations may assist in the development of individualized pharmacotherapy avoiding unexpected drug responses, such as harmful adverse drug reactions or treatment failures. This review discusses both the basic characteristics of the major polymorphic cytochrome P450 enzymes and examines the pharmacogenetic methods employed to estimate metabolic status. We will focus mainly on the basic principles of genotyping assays involving molecular biology tools. PMID- 17402552 TI - The effects of orlistat treatment interruption on weight and associated metabolic parameters. AB - Due to the insurance companies' restrictions, partial reimbursement of orlistat treatment in the Czech Republic is restricted to obese diabetics with BMI >35 who are concurrently treated pharmacologically for dyslipidaemia, hypertension or ischaemic heart disease, with compulsory interruption of minimum 3 months, only after which the treatment can be resumed for another year. We evaluated 32 patients with Type 2 diabetes who underwent such course of treatment, with view of establishing whether the interruption has any detrimental effect on the success of the therapy in terms of weight loss and diabetes compensation. The treatment was well tolerated, producing statistically significant decrease in BMI and triglyceride levels during the first year, which was maintained in the second year. Fasting glucose levels were improved at nearly-significant level. The interruption in treatment between the first and second year had no marked detrimental effect, although the relative failure of the second treatment year to bring further benefits to the patients can certainly be at least partially attributed to this treatment gap. PMID- 17402553 TI - Progestagens androgenic action on the bone of male castrated mice. AB - It has been suggested that some progestagens could have an androgenic stimulatory effect on bone formation. The androgenic effects of progestagens were tested in vivo in the absence of androgens and estrogens in the castrated male mice, species extraordinary responsive to the withdrawal or administration of androgens. Three progestagens (norethisterone, uterogestan and medroxyprogesterone acetate were compared as to their androgenic activity. Tissues especially sensitive to androgens, the seminal vesicles and kidney of the mice fell significantly after castration and all three progestagens did not affect their weights. The present results confirm the well known fact that castration leads to osteopenia in mice. Uterogestan micronized progesterone and MPA have no effect on the bone density or mineral content of the tibia of tested mice. Only in the case of NETA we observed slight statistically significant (p < 0.05) increase in bone density. Progestagens do not appear to have the androgenic effect on the skeleton and NETA has been suggested as one of the exception. Our results indicate that only NETA at the dose which is used in hormonal therapy for prevention of osteoporosis has a slight protective effect against bone mineral loss in castrated mice. PMID- 17402554 TI - Kainic acid and nitrergic neurons in immature hippocampus. AB - Using histochemical analysis the effect of intraperitoneal administration of kainic acid on hippocampal neurons was studied. 18-day-old male rats of the Wistar strain received kainic acid (10mg/kg) in one dose. Two days later, the 20 day-old animals were transcardially perfused with 4% paraformaldehyde under deep thiopental anaesthesia. Cryostat sections were stained to identify NADPH diaphorase positive neurons that were then quantified in the CA1 and CA3 areas of the hippocampus, in the dorsal and ventral blades of the dentate gyrus and in the hilus of the dentate gyrus. Combination of the Fluoro-Jade B and bis-benzimide (Hoechst 33342) was used in the same areas, to identify possible neurodegeneration. After the kainic acid administration the number of NADPH-d positive neurons was lower in CA1 and CA3 areas of the hippocampus and in the hilus of the dentate gyrus, compared to the control group which we consider as baseline. Fluoro-Jade B staining detected a moderate density of neurodegeneration after KA administration in CA3 and CA1 areas of the hippocampus and the hilus of the dentate gyrus. PMID- 17402555 TI - Neuroprotective effect of (R, S)-4-phosphonophenylglycine against neuronal damage associated with homocysteic acid-induced seizures in immature rats. AB - Incidence of human epilepsy in infants and children is high and prolonged seizures in the early developmental period can cause brain damage and lead to serious consequences later in the life. The present study was aimed to investigate potential protective effect of (R, S)-4-phosphonophenylglycine ((R, S)-PPG), a potent and selective group III mGluR agonist, on brain damage associated with homocysteic acid-induced seizures in immature 12-day-old rats. This compound does not exhibit any proconvulsive effect. Moreover, (R, S)-PPG was shown to protect NMDA and quinolinic acid-induced lesions in rats. Seizures were induced by bilateral intracerebroventricular (i.c.v.) infusion of homocysteic acid (DL-HCA, 600 nmol/side). (R, S)-PPG was given by bilateral i.c.v. infusions (5 nmol/side) at 15- to 20-min time intervals prior to administration of DL-HCA. After 1 or 6 days of survival, animals in all experimental groups (13-day-old and 18-day-old) were perfused transcardially under deep ether anaesthesia with heparinized normal saline and subsequently with the fixation solution (4% paraformaldehyde in the phosphate buffer, pH 7.4, both solutions at room temperature). Two histological methods were used in our study. Fluoro-Jade B dye is an anionic fluorescein derivative useful for the histological staining of neurons undergoing degeneration and staining with bis-benzimide (Hoechst 33342) was used to detect apoptotic cells according nuclei with condensed and/or fragmented DNA. Animals perfused 1 day after the treatment (13-day-old): After only (R, S)-PPG application, no obvious pathological changes were found. After only DL-HCA application, distinct destruction of the hippocampal region both in the dorsal and ventral hippocampus was observed. Particularly affected were cells in the CA1 and CA3 regions. In addition, neurons with segmented or fragmented nuclei were found in the granule cell layer of the dentate gyrus. (R, S)-PPG + DL HCA administration resulted in a lower number of Fluoro-Jade B positive cells. All areas of the hippocampus were protected by (R, S)-PPG pre-treatment. Animals perfused 6 days after the treatment (18-day-old): In the group where only (R, S) PPG has been applied, no obvious pathological changes were found in the hippocampal area. After only DL-HCA administration almost complete destruction of the hippocampal region both in the dorsal and ventral hippocampus was observed. Particularly affected were the cells in the CA1 and CA3 regions, granule cells of the dentate gyrus and many interneurons in all hippocampal areas. (R, S)-PPG + DL HCA administration resulted in lower number of Fluoro-Jade B positive cells. All areas of the hippocampus have been protected by (R, S)-PPG pre-treatment. In conclusion, the present data support the hypothesis that (R, S)-PPG can have a beneficial effect in those disorders where excitotoxicity is one of the dominant pathogenetic mechanisms. PMID- 17402556 TI - Use of stabilometric platform and visual feedback in rehabilitation of patients after the brain injury. AB - Rehabilitation of patients after the brain injury requires employing of all available mechanisms of neuroplasticity. To achieve it, the voluntary activation of brain systems that are involved in the signal processing, represents the most effective tool. The control of balance is a complex neuronal mechanism based on unconditioned and conditioned reflexes, as well as on the actual cognitive processes. As it requires participation of several brain regions, training of the posture support mechanisms can provide a highly effective tool for rehabilitation. The aim of the study was to develop methods for the long-term follow up and training of the balance skills in patients with different types of brain impairment. To obtain standard data, the stabilometric platform Posturograph STP-03 and special examination programs were also used in the study of the equilibrium skill training by healthy volunteers. For the assessment of the learning efficiency two criteria from the recorded data were used - the velocity of adjustment of the gravity centre and the accuracy of the movements. Stabilometric platform was used also for the balance skill training with the visual biofeedback. Our results show that the proposed program for the equilibrium skill training offers a comparatively simple method of the adequate duration with numerical and graphical output, which allows fast interpretation of the treatment results. The synoptic form of results can also stimulate the patient's motivation during the long-term training for the mobility improvement. PMID- 17402557 TI - Repetitive miscarriages in a patient with a high level of anticardiolipin antibodies and myocardial infarction. AB - Antiphospholipid antibodies consist of a group of heterogeneous autoantibodies against anionic phospholipids. We describe the case of a 19-year-old patient who was consulted in the obstetrics service after her second miscarriage. She was a smoker and six months after this consultation she developed diabetes mellitus. At 21 years of age she suffered from myocardial infarction, when high anticardiolipin antibody levels were evidenced. Some months later the patient again became pregnant and prophylaxis against miscarriages was performed using low doses (5000 IU) heparin administered subcutaneously at 12-hour intervals. She evolved with preeclampsia, however, the baby was born in good health. One year after she again became pregnant and prophylaxis against miscarriage was again performed using low doses of heparin. The pregnancy successfully resulted with the birth of her second child. PMID- 17402558 TI - Science in the service of occupational health: the case of the commission for "miner's disease of Jachymov" in the inter-war Czechoslovakia. PMID- 17402559 TI - [The eye and aging]. PMID- 17402560 TI - [Translational research with experimental autoimmune uveoretinitis (EAU)]. AB - Experimental autoimmune uveoretinitis (EAU) induced by immunization with retinal antigen (Santigen or interphotoreceptor retinoid-binding protein; IRBP) serves as an animal model of human uveoretinitis. As the first stage, we demonstrated the similarities between EAU and ocular inflammation in Behcet's disease by investigating anti-retinal antibodies, leukocyte migration inhibition by retinal antigen, immunogenic antigens, aberrant functions of neutrophils, and dominant Th1 lymphocyte reaction. From these findings, we verified that EAU, which is not associated with the systemic disorders observed in Behcet's disease, is an appropriate model for translational research targeting ocular inflammation. In the second stage, we set 3 therapeutic strategies for uveitis in Behcet's disease to be conducted in the translational research: (1) intraocular administration of an immunosuppressive drug; (2) inhibition of Th1 lymphocytes; and (3) activation of immunoregulatory cells. In strategy 1, our studies indicated that intravitreal injection of 10 microg of tacrolimus (FK 506) was not harmful to the retina and was predominantly effective in suppressing ongoing EAU in rats. In strategy 2, two approaches were adopted to prevent differentiation of Thl cells. One is anti cytokine antibody therapy using anti-IL-12 monoclonal antibodies(mAb). The other is blockade of co-stimulatory signals, especially the ICOS-B7RP-1 pathway. Administration of anti-IL-12 mAb at the time of IRBP immunization completely inhibited development of EAU, and antagonistic anti B7RP-1 mAb suppressed the severity of EAU even when administered after development of EAU. In strategy 3, adoptive transfer of antigen presenting cells treated with a neuropeptide (vasoactive intestinal peptide or calcitonin gene-related peptide) or CD 4+ CD 25+ regulatory T cells suppressed EAU. We look forward to the day when therapies that are being developed in our translational research using EAU will become available for treating intraocular inflammation in Behcet's disease. PMID- 17402561 TI - [Physiology and pathology of visual information processing]. AB - (1) Human crystalline lens and vision In order to investigate the hue changes in eyes with UV-absorbing intraocular lenses (IOLs) and tinted IOLs, we simulated the changes in the chromaticity coordinates of the 16 colors of the Farnsworth dichotomous test-panel D-15 (panel D-15), considering the ratio of the spectral transmittance of the IOL and the human crystalline lens, and the results were plotted on a CIE chromaticity diagram. The chromaticity coordinates of each color for UV-absorbing IOLs shifted to close to the origin of coordinates while retaining their hue circle. However, the chromaticity coordinates for the eyes with tinted IOLs did not change much compared to the coordinates for phakic eyes. As a result, it was suggested that cyanopsia after UV-absorbing IOL implantation could be explained by this simulation. As far as the color perception is concerned, it was also felt that tinted IOLs were superior to UV-absorbing intraocular lenses. Next, in order to evaluate the hue changes after IOL implantation, the achromatic point settings were measured once before surgery and several times at intervals after surgery after taking off the eyepatch. Four subjects participated in the experiments. There was a large shift into the "yellowish" region of color space immediately after taking off the eyepatch after cataract surgery. Then, the achromatic point returned to the chromaticity near the achromaticpoint measured prior to the surgery, with the time course of a long time, compared to color constancy in our daily life, which takes as long as several hundreds of seconds to reach an asymptote. Therefore, the mechanism of achromatic point shifts after cataract surgery may be different from the color constancy mechanism in everyday life. (2) Molecular genetics and vision We demonstrated new clinical and genetic aspects of congenital red-green color vision defects, congenital achromatopsia, enhanced s-cone syndrome (ESCS), and Oguchi disease in Japanese patients. We clinically diagnosed 88 male dichromats(31 protanopes, 56 deuteranopes, and one unclassified subject). This subject had a new form of X-linked pigment gene with a unique arrangement of exon 5(Y277 from the long-wavelength-sensitive gene and A 285 from the middle wavelength-sensitive gene). Mutational analysis of patients with achromatopsia disclosed CNGA3 mutations (p.R 436 W, p.L633 P) in one of 14 patients, suggesting low frequency (7%, 1/14) of CNGA3 mutations in the Japanese population. Three novel NR2E3 mutations (p.R 104 Q, p.R 334 G, p.Q 350 X) were identified in both mild and severe forms of ESCS. A novel homozygous GRK1 mutation (p.P 391 H) was found in the Oguchi disease patient with reduced cone responses. This is the first reported Japanese patient with GRK1 -associated Oguchi disease. 3. Information processing of the visual cortex and vision Regarding information processing in the visual cortex, we developed the stimulus to improve identifying retinotopy of the human visual cortex. We performed two types of fMRI experiments. One provided a quick method of mapping retinotopy using a composite stimulus with both ring- and wedge-shaped stimuli. The other provided a method which can show the horizontal meridian clearer. We explored the activation of the visual cortex associated with color perception. In our studies of the color center, we first researched the symptoms and lesions of cerebral achromatopsia, and we next performed the fMRI experiments with a pseudoisochromatic plate test and with a color arrangement test. After this we also performed the fMRI experiments with a complex color painting. We realized objective perimetry with functional brain images. We first developed the software to depict a visual field from the signals of MR imaging. Next we performed the experiment with hemifield stimulation and showed the possibility of its clinical application. Then we showed its reproducibility, performing the experiment with more complicated letter-shaped masked visual stimulation. Finally, we applied the technique to patients with cerebral dysfunction. We performed diffusion tensor imaging (DTI) with a clinical 1.5 T MR machine to visualize optic radiation. With patients who were clinically expected to show disorder of optic radiation, these visualizations were consistent with their pathologies. It was suggested that this new DTI technique is useful for estimating functional disorder of optic radiation. PMID- 17402562 TI - [Oxidative stress and inflammation: hypothesis for the mechanism of aging]. AB - Oxidative stress due to free radicals is related to the pathogenesis of many chronic disorders including cancer, inflammation, and neurological diseases. Oxidative stress such as aging and light exposure is also considered to be associated with age-related macular degeneration and cataract. The ocular surface is chronically exposed to oxidative stress including ultraviolet light, the oxygen in air, and changes in oxygen pressure due to blinking. We demonstrated that a rat dry eye model with a jogging board showed corneal epithelial disoders and elevated levels of oxidative stress, suggesting that the pathogenesis of epithelial disorders in dry eye with low frequency of blinking is related to oxidative stress. Next, using a model of laser-induced choroidal neovascularization (CNV), we showed that angiotensin receptormediated inflammation is required for the development of CNV. We also demonstrated that mice deficient in superoxide dismutase (SOD) showed typical clinical features of AMD. Finally, we proposed our thoughts about regenerative medicine, that is, to maintain quiescent stem cells, we have to regulate the aging of stem cells. PMID- 17402563 TI - [Aging and retinal vascular diseases]. AB - Ocular vascular diseases such as diabetic retinopathy, retinal vein occlusion, and age-related macular degeneration, whose population increases along with aging, have become leading causes of severe visual disturbance. Macular edema and serous retinal detachment are associated with abnormal vascular leakage and tractional retinal detachment, and neovascular glaucoma is caused by retinal neovascularization. Such ocular vascular diseases are caused by vascular cell aging and vascular damage associated with lifestyle-related diseases including diabetes mellitus, hypertension, hyperlipidemia, and obesity. In the present study, we investigated molecular mechanisms in such vascular deficiencies using vascular cell biology methodology, and we propose novel strategies for the treatment of such vascular diseases. Along with aging, oxidative stress and physical stress, such as mechanical stretch, continuously and directly insult vascular cells. Such stress induces apoptosis by intracellular signaling through stress kinases in cultured retinal vascular cells. Inhibition of such stress kinases could be an effective treatment to protect the vascular cells against age related damage. In a retinal vascular developmental model, pericyte loss causes pathology mimicking macular edema and proliferative diabetic retinopathy. Angiopoietin 1 (Ang 1) secreted by pericytes suppresses oxidative stress-induced intracellular signaling through stress kinases linked to cell apoptosis and normalizes such retinal pathology. This suggests that the paracrine action of Ang 1 in the pericytes is necessary to sustain normal retinal vasculature, and that Ang 1-triggered intracellular signaling is useful for the treatment of vascular cell pathology associated with pericyte loss. In diabetic retinopathy and retinal vein occlusion, retinal vessels regress along with retinal vascular cell apoptosis, and the retina becomes ischemic followed by pathological retinal neovascularization. VEGF has been recognized as a predominant factor to induce the ischemic retinal neovascularization. We found that retinal vascular cells have a characteristic pattern in VEGF receptor expression, which causes vascular pathology more frequently in the retina than in other organs. Neuropilin 1 (NRP 1), which enhances VEGF receptor function, is abundantly expressed in the retinal endothelial cells and is upregulated by VEGF itself and by hypoxia to regulate a positive feedback mechanism in retinal neovascularization. This receptor could be a unique target for retina-specific therapy. Lifestyle-related diseases increase along with aging and have further increased due to changes in Japanese lifestyle imitating that of Western countries. We found that the renin-angiotensin system which regulates hypertension and cardiovascular diseases, and adipocytokines which are abnormally secreted in obesity, act as proangiogenic factors. Regulation of such lifestyle-related disease factors is important for the treatment of retinal vascular diseases. Finally, we found that erythropoietin is an ischemia-induced angiogenic factor that acts independently and as potently as VEGF in proliferative diabetic retinopathy (PDR). Our study utilizing human vitreous samples demonstrates that the VEGF level is particularly high and strongly associated with angiogenic activity in PDR patients. The potential of VEGF inhibitors has recently been recognized in clinical applications. The manipulation of each angiogenic factor and adipocytokine that we report here could become potential therapy in the near future. PMID- 17402564 TI - [Novel approach for management of age-related macular degeneration- antiangiogenic therapy and retinal regenerative therapy]. AB - Age-related macular degeneration (AMD) is a leading cause of legal blindness in developed countries. Even with the recent advent of several treatment options, treatment of exudative AMD, characterized by choroidal neovascularization (CNV), remains difficult. Thus, in this review article, we report on the investigation of novel approaches for the management of AMD, antiangiogenic therapy for CNV, and retinal regenerative therapy. Polyion complex(PIC) micelles have a range in size of several tens of nanometers formed through an electrostatic interaction, and accumulate in solid tumors through an enhanced permeability and retention(EPR) effect. In this study, we examined the distribution of the PIC micelles which encapsulate fluorescein isothiocyanate-labeled poly-L-lysine{FITC P(Lys)} in experimental CNV in rats, to investigate whether PIC micelles can be used for the treatment of CNV. We demonstrated that PIC micelles accumulate in the CNV lesions and are retained in the lesions for as long as 168 hours after intravenous administration. These results raise the possibility that PIC micelles can be used for achieving an effective drug delivery system against CNV. Although photodynamic therapy (PDT) is a very promising treatment for AMD, most patients require repeated treatments. For effective PDT against AMD, the selective delivery of a photosensitizer to the CNV lesions and an effective photochemical reaction at the CNV site are necessary. The characteristic dendritic structure of the photosensitizer prevents aggregation of its core sensitizer, thereby inducing a highly effective photochemical reaction. We present an effective PDT for AMD employing a supramolecular nanomedical device, i.e., a novel dendritic photosensitizer encapsulated in a polymeric micelle formulation. With its highly selective accumulation in CNV lesions, this treatment resulted in a remarkably efficacious CNV occlusion with minimal unfavorable phototoxicity. Our results will provide a basis for an effective approach to PDT for AMD. Spatial control of gene transfection in the body is a core issue in the gene therapy for ocular diseases including AMD. Photochemical internalization (PCI) is a technology that effects light-induced delivery of DNA directly inside cells. PCI usually requires that a photosensitizer be added to the drug-delivery system to photochemically destabilize the endosomal membrane. We have developed a ternary complex composed of a core containing DNA packaged with cationic peptides and enveloped in the anionic dendrimer, phthalocyanine, which provides the photosensitizing action. Subconjunctival injection of the ternary complex followed by laser irradiation resulted in transgene expression only in the laser-irradiated site in rats. This PCI-mediated gene delivery system is potentially useful in gene therapy for ophthalmic diseases. Accumulation of lipofuscin is related to an increased risk of AMD. We report that a major lipofuscin component, A2E(N-retinyledin-N retinylethanolamin), activates the retinoic acid receptor (RAR). In vivo experiments suggest that A2E accumulation results in the pro-angiogenic conversion of retinal pigment epithelial(RPE) cell phenotype. This physiological consequence of A2E accumulation may be related to a novel potential therapeutic target for CNV. To recover visual function damaged by AMD, retinal regenerative therapy is essential. We investigated whether subretinal transplantation of bone marrow mesenchymal stem cells(MSCs) promotes photoreceptor survival in a rat model of retinal degeneration. Morphological and functional studies in vivo, including histological analysis and electrophysiological studies, indicate that the subretinal transplantation of MSCs delays retinal degeneration and preserves retinal function. These results suggest that MSC is a useful cell source for cell transplantation therapy for retinal degeneration. In order to elucidate the molecular mechanisms of development of the fovea, which is composed mainly of cone photoreceptors and is susceptible to injury from AMD, we performed a comparative gene expression analysis between the central and peripheral regions of the monkey retina using monkey (rhesus macaque) genome microarray chips. We then selected the clones which were expressed at significantly higher levels in the central region and confirmed their expression in the monkey retina by section in situ hybridization. This study sheds light on the mechanisms of foveal development and may lead to the development of regenerative medicine for cone photoreceptors. PMID- 17402565 TI - [Number of bibliographies cited from the Japanese Journal of Ophthalmology]. PMID- 17402566 TI - [The analysis of the state and the dynamics of the health manpower resources of the subjects of the Russian Federation in 1990 - 2004]. AB - The labor and manpower resources are among the major resources of public health system. The article presents the analysis of the provision of the population of the Russian Federation with the medical and paramedical personnel during the last fifteen years both on the national and regional level. The revealed territorial differences in resources provision and two- and three-fold abmodality of the indices of medical and paramedical personnel supply testify of the urgent need in the comprehensive systemic approach in resolving the problem of the availability of the high-quality medical care to the population. PMID- 17402567 TI - [The monitoring of the congenital malformations development in children: problems and prospects]. AB - In various countries all over the world, the congenital malformations are among the leading causes of infant mortality and children disability. The historical approach is applied to consider epidemiological monitoring as a practice preceding the comprehensive analysis of causes and conditions of congenital malformations formation. The primary tasks of monitoring study are established and means of their implementation as well. The actual technique of registration and recording of various forms of the congenital malformations in children and its alternative are presented. The need of the coordinated activities of pediatric, medical genetic and epidemiological services in the organization and the execution of the monitoring in different regions of the country is emphasized. PMID- 17402568 TI - [The impact of the mode of life on health]. AB - On the basis of the Rosstat data and the results of numerous medical sociological studies, the causes of the prevalence in the Russian Federation of such adverse habits as alcoholism and tobacco smoking which negatively affect the health level of various population groups are analyzed. PMID- 17402569 TI - [The social hygienic risk factors during the involutional processes in males]. AB - The issues related to the involutional period in males are discussed. It is demonstrated that the development of complicated involutional period is under considerable impact of not only genetic factors but such socially important factors as alcoholism in family, nicotine dependence of mother or father, pecuniary component in family, etc. The malignancy of the combined effect of biogenetic, exogenous and acquired factors is verified. PMID- 17402570 TI - [On the methodology of evaluation of the abiotic environmental risk factors impacting population health]. AB - In the contemporary conditions of civilization progress, the population health is considered as a key backbone factor of national security. The development of the methodology of evaluation of risk factors for population health is important both for the strategy of management of human environment, a new integrating guideline of contemporary medical science--environmental medicine and for optimization of social hygienic monitoring with the view of providing the epidemiological well being of population. Based on the synergistic approach, the algorithm of evaluation of abiotic risk factors for population health applying the equations of partial derivatives in such distributed natural systems as the environment. PMID- 17402571 TI - [The impact of the social economic conditions on the reproduction processes]. AB - The social economic conditions of population reproduction in the Trans-Dniester Region with the emphasis on its specifics are analyzed. The natural dynamics of population is estimated including gender and age peculiarities, marriage and divorces statistics, migration processes and population health in the region. It is shown that the population reproduction dynamics follows the natural laws and processes. The role of the post-Soviet period social economic conditions harmful to health are considered. The interdependencies between the social economic development and population reproduction are revealed. The recommendations related to the means of the further enhancement of social economic conditions as related to population reproduction in the Trans-Dniester Region are proposed. PMID- 17402572 TI - [The capacity and the pattern of the ambulatory polyclinic care provided by the general practitioners and the district doctors]. AB - In the Republic of Dagestan, the capacity and the pattern of the ambulatory polyclinic care provided to urban population were studied during 2004-2005 in four cities--Makhachkala, Derbent, Buynaksk and Kizyljurt. The structure of 2465 visits to the general practitioners (family doctors) and 2549 visits to the district doctors was studied. In the structure of appealability to the general practitioners (family doctors) 30% fall to the neurological patients. The general practitioners (family doctors), as compared with the district doctors, twice as much pay attention to preventive activities (16.2% vs. 8.8%) and they much more often appoint their own patients to subspecialists' consultation (93.1% vs. 85.2%). It was established that 53.6% of patients with ischemic cardiac disease and 45% of patients with diseases of digestive system consult the doctor at the eighth day from the beginning of disease and later. The research data gave an opportunity to elaborate the guidelines and suggestions related to the optimization of the ambulatory polyclinic care provided by the general practitioners (family doctors) and district doctors to urban population. PMID- 17402573 TI - [The physicians' awareness of urogenital system tuberculosis]. AB - The chair of public health of the Omsk State medical academy conjointly with the municipal TB dispensary No 4 carried out the medical social study of the urogenital system tuberculosis since no profound and research of this problem was organized on the territory of Omsk oblast. The article presents the fragment of the data related to the survey of he physicians of medical institutions form city of Omsk and the Omsk oblast about the effectiveness of the medical social care to the patients suffering from the urogenital system tuberculosis. PMID- 17402574 TI - [The prevention of tuberculosis among migrants]. PMID- 17402575 TI - [The problems of the development of rehabilitation technologies]. AB - The research in the field of theoretical methodological problems of preservation and recovery of population health on the basis of studying the impact of the social and biological factors is high-top priority guideline of the strategy and tactics of health care in enhancing the medical social care. The actuality of consideration in the Russian Federation the concept of the rehabilitation care to people with vital activity restrictions depends on many reasons. The patients with stoma consist a specific group of persons survived severe disease or trauma or very often mutilating "stomic" operation which entailed the above-mentioned vital activity restrictions. Among people with intestine or ureter stoma , the disability substantially differs from any other disability developed due to other causes. The rehabilitation as an active form of the social protection is to be directed towards preserving the quality of life of "stomised" patients and changing their social position i.e. engaging into community. It is assumed that the community adaptation to the specific needs of such category of disabled persons and environment alteration are the most necessary components of the medical social rehabilitation. PMID- 17402576 TI - [On some aspects of the practice of the pathologoanatomic services]. AB - It is demonstrated that the adequacy of the data of the structure of population mortality depends not so much upon the indicators of the practice of the pathologoanatomic services as upon the commitment both of the authorities and general community. PMID- 17402577 TI - [On the mistakes during the completion of the medical certificate of death]. AB - The article deals with the typical mistakes made by physicians during the completion of the medical certificate of death. To enhance the reliability of mortality statistics such means as canvass of mistakes and application of the new technologies (the software package "The monitoring of natality and mortality" and the courseware "RUTEN-DON") are proposed. PMID- 17402578 TI - [The judgments of the experts on the means of optimizing the private medical practice]. PMID- 17402579 TI - [The prevention of the primary disability over the cardiovascular diseases in the Republic of Kazakhstan]. AB - The disability is the conclusive stage of the development of major cardiovascular diseases. The prevention of the primary disability because of this pathology includes early identification of the disease, timely and exact assessment of the working capacity, the personal commitment of the patient in soonest restoration of working ability, the cooperation of different institutions, agencies and authorities in an effort to decrease the morbidity and disability. PMID- 17402580 TI - [The base hospitals during the Great Patriotic War in the Southern Ural]. PMID- 17402581 TI - [The approaches in the formation of zemstvo and urban medicine]. PMID- 17402582 TI - [The involvement of G.Y. Rhein in the formation of the Russian forensic medical expert service]. PMID- 17402584 TI - [Effects of noopept and cortexin on the behavior of matured rats treated with corticoliberin or 70-kDa heat shock proteins in early ontogeny]. AB - Young Wistar rats aged 4 days were injected intraperitoneally with corticotropin releasing hormone (CRH), which is an agent activating the stress system, or 70 kDa heat shock proteins (HSP-70)--intracellular shaperons, possessing antistress properties. In grown adult rats aged 90-100 days, the effects of nootropic drugs noopept and cortexin (1 mg/kg, i.p.) were assessed. The activation of stress or antistress systems with CRH or HSP-70 significantly altered the drug action. The effects were different in males and females and depended on animal gender. The spectrum of pharmacological activity of noopept and cortexin changed: noopept demonstrated preferable psychoactivating and antiaggressive effects, whereas cortexin showed mild anxiolytic and antidepressant activity. It is suggested that the behavioral effects of nootropes depend on the conditions of the stress system formation in early ontogeny. PMID- 17402583 TI - [Comparative study of the effects of amitriptyline, fluoxetine, and tianeptine on the amplitude of transcallosal evoked potentials]. AB - The influence of amitriptyline, fluoxetine and tianeptine upon transcallosal responses has been studied in male albino rats. It is established that amitriptyline does not influence, fluoxetine reduces, and tianeptine increases the amplitude of evoked potentials. These data show evidence that tianeptine facilitates, fluoxetine impairs, and amitriptyline does not affect the interhemispheric transmission. PMID- 17402585 TI - [Comparative study of the electrophysiological properties of class iii medicinals (cardiocyclide, nibentan, sotalol) and evaluation of their efficacy in atrial fibrillation produced by auricular and vagal stimulation]. AB - The electrophysiological properties of cardiocyclide, nibentan and sotalol and their efficacy on a model of atrial fibrillation were compared in experiments in anaesthetized dogs. The electrophysiological parameters of drugs were investigated using the method of programmed electrical stimulation of myocardium on the background of excitation of the peripheral segment of the right vagus nerve with current pulses of increasing frequency. The atrial fibrillation was produced by short stimulation (10 pulses) of the right auriculum (10 Hz, 4 thresholds). Cardiocyclide was injected in a dose of 2.5-5.0 mg/kg; nibentan, 0.25 mg/kg; and sotalol, 2.5 mg/kg. Cardiocyclide was found to prevent vagotonic atrial fibrillation in 80% of cases; under vagal stimulation this drug maintained the electrophysiological attributes and exhibited the frequency-independent action inherent in this agent. Nibentan completely retained its ability to prolong the ventricular repolarization under vagal stimulation conditions, increased the effective atrial and ventricular refractory periods, and showed the ability to prevent and eliminate the vagotonic atrial fibrillation in 80-90% of cases. Under vagal stimulation, sotalol increased the repolarization and prolonged the effective refractory atrial and ventricular periods, reduced the heart rate, and suppressed the sinus node function. The action of sotalol was frequency-dependent. Sotalol was found to prevent the vagotonic atrial fibrillation in 60% of cases. PMID- 17402586 TI - [Comparative study of the antiarrhythmic activity of L-, D-and DL-stereoisomers of potassium magnesium aspartate]. AB - Injection forms of potassium (K) and magnesium (Mg) aspartate (Asp) were compared in preventing cardiac disorders caused by electrolytic disturbances, primarily low K and Mg levels (e.g. caused by the treatment with cardiac glycosides and diuretic drugs). Widely used K- and Mg-Asp preparations (asparkam, panangin, pamaton) are synthesized from aspartic acid representing a racemic mixture of L- and D-stereoisomers. Differences in metabolism and utilization of D- and L-amino acids probably influence the pharmacological properties of K and Mg L- and D aspartates. Moreover, the pharmacologically effective doses of Mg and K salts can induce toxicity, which depends on the nature of anions. The aim of this study was to compare of antiarrhythmic action of K and Mg L-, D-, and DL-Asp stereoisomers using calcium chloride (CaCl2) and aconitine induced arrhythmia models in rats and strophanthin-K induced arrhythmia model in guinea pigs. It was found that intravenously administered K- and Mg-L-Asp exhibited higher activity compared to K- and Mg-D- and DL-Asp on the strophanthin-K, CaCl2, and aconitine induced arrhythmia models. Indeed, K- and Mg-L-Asp more effectively decreased the incidence of arrhythmias, increased the time to onset of the first arrhythmia, decreased percentage loss of rats, and increased the survival life of animals after the first arrhythmia onset in rats with arrhythmias induced by strophanthin K and CaCl2 as compared to K and Mg-D- and DL-Asp. At the same time K- and Mg-L Asp was better than D- and DL-Asp with respect to acute toxicity (LD50), effective dose (ED50) and antiarrhythmic (therapeutic) ratio (LD50/ED50) in rats with aconitine-induced arrhythmia model. PMID- 17402587 TI - [Antiarrhythmic activity of lidocaine under conditions of lipid and carbohydrate metabolisms disturbances in modeling early occlusional and reperfusional arrhythmias]. AB - The antiarrhythmic activity of lidocaine on the models of early arrhythmias induced by occlusion and reperfusion was studied in rats with background derangements of lipidic and carbohydrate metabolisms. It was established that lidocaine in a dose of 100 mg/kg did not produce reliable antiarrhythmic action under these conditions. In addition, changes in the development of early occlusional and reperfusional arrhythmias against the background violation of lipid and carbohydrate metabolisms were found. PMID- 17402588 TI - [Cytoprotective properties of 3-oxy-6-methyl-2-ethylpyridine succinate in cell cultures]. AB - Tests on calf coronary vessel cells (CVC) cultures showed that 3-oxi-6-methyl-2 ethylpiridine succinate (OMEPS) in dozes within 5.0-10.0 microg/ml did not decrease proliferation index, monolayer development, and viability factor. OMEPS also did not increase cytotoxicity index and did not change cytomorphological characteristics of the culture. In a concentration of 100 microg/ml, OMEPS exhibited some cytotoxyc properties, but to a lower degree compared to the reference antiarrhythmic drug moricisine (1.0-20.0 microg/ml). However, OMEPS (2.5 microg/ml) reduced the cytotoxicity moricisine introduced into CVC culture in a doze of 1.0 microg/ml. PMID- 17402589 TI - [Effect of dry aspen bark extract on the motor function of gastrointestinal tract in mice]. AB - Experiments on mice (both intact and those with reserpine induced gastric ulcers) showed that dry aspen bark (DAB) extract stimulated the motor gastric activity. The DAB preparation produced a moderate purgative effect. PMID- 17402590 TI - [Effect of exogenous thyroxin on the state of kidney transport of nitrites and nitrates in rats]. AB - The transport of nitrites and nitrates in the kidney has been studied on the background of single and prolonged injection of thyroxine. The function of kidneys and the state of renal transport of endogenous nitrites and nitrates were studied depending on the duration of thyroxin administration alone and in combination with an unspecific inhibitor of NO-synthases (Nomega-NLA) or sodium nitrite. It was established that thyroxin injection results in a decrease in the glomerular filtration rate, increase of proteinuria, and enhanced excretion of nitrite and nitrates via kidneys already in 2 h after hormone administration. After a 7-day injection of the hormone, the rate of glomerular filtration was retained on a reduced level, and the excretion of protein by the kidneys exceeded the control values. At the same time, on the background of increased concentration of nitrites in the blood plasma, the excretion of nitrites via kidneys (per 1 ml glomerular filtrate) was lower and the excretion of nitrates was substantially higher than in the control. In an acute period, the injection of thyroxine stimulated the production of nitrites and nitrates in the organism, which was accompanied by an increase in their concentration in the plasma and by enhanced excretion of these substances via kidneys. In the case of long-term injection of thyroxine, a decrease in the clearance of nitrites is among the basic mechanisms that provide the retention of nitrites in the extracellular liquid. PMID- 17402591 TI - [Transplacental effects of bemithyl]. AB - The daily administration of bemithyl (20 mg/kg) from 6 th to 16 th day of pregnancy in female rats led to the decrease in fetal death after the implantation and increased fetal body weight. The treatment of pregnant rats also led to acceleration of the development of physical condition and sensomotor reflexes of progeny in the postnatal period. PMID- 17402592 TI - [Effect of nebivolol on the blood oxygen transport]. AB - The effect of nebivolol (30.0, 60.0, or 90.0 microM) on the blood oxygen transport was studied by incubation with blood samples for 120 min. Nebivolol increased the values of p50 (pO2 at 50% hemoglobin saturation with oxygen) under real pH and pCO2 (by 4.3 +/- 0.8 mm Hg at the lowest concentration (p < 0.01). The further (2- or 3- fold) growth in the concentration of nebivolol increased these values by 7.5 +/- 1.1 and 10.6 +/- 0.7 mm Hg, respectively (for both p < 0.01), thereby indicating the dose dependence of the drug effect. Greater drug concentration resulted in higher methemoglobin level and nitrate/nitrite content. The data can be explained by a change in the hemoglobin-oxygen affinity as a result of the influence of nebivolol through an autonomous intraerythrocyte system that regulates the blood oxygen-binding properties. In this case, NO acts as an important modifier of the hemoglobin function. PMID- 17402593 TI - [Melatonin as a corrector of free radical oxidation processes during toxic damage of liver in rat]. AB - Biochemiluminiscense parameters (light sum and maximum flash) show that free radical oxidation intensity (increased in serum blood of rats with toxic hepatitis) is decreased upon the treatment with melatonin. At the same time, the tg a2 value (reflecting the total antioxidant activity) changes toward the normal level, which indicated a decrease in the degree of antioxidant system mobilization. Superoxide dismutase and catalase activities (increased in the liver and blood serum of rats with toxic hepatitis) also showed the tendency to decrease upon the melatonin administration. Evidently, melatonin can act as a factor correcting the oxidative stress caused by the toxic damage of liver. PMID- 17402594 TI - [Features of expression and functional activity of P2X7 receptors in bone marrow cells under the action of doxorubicin]. AB - Mechanisms of the myelotoxic action of doxorubicin associated with changes in the expression and functional activity of P2X7 receptors have been assessed. The acute and subacute exposure of bone marrow cells to doxorubicin in vivo changed the expression of P2X7, altered the intracellular and extracellular ATP concentrations, and modulated the process of programmed cell death. These changes were associated with transformed susceptibility of hemopoietic cells to the apoptogenic action of ATP. Various possible mechanisms of realization of the apoptogenic action of ATP during acute and subacute exposure to doxorubicin are discussed. PMID- 17402595 TI - [The effect of rubomycin, mexidol and emoxypine on some metabolic indexes and the proccess of spontaneous metastasis under conditions of experimental neoplasia]. AB - The effects of mexidol and emoxypine on some indexes of endotoxicosis and weighted characteristics of tumor carrier during experimental neoplasia have been studied under the conditions of therapy using antracycline antibiotic rubomycin. PMID- 17402596 TI - [Development of acute exudative inflammation in rats upon perftoran infusion]. AB - The effect of the perftoran--a blood replacing emulsion with multifunctional properties--on the biochemical parameters of blood serum has been studied in rats with acute exudative inflammation. It is established, that perfloran does not influence the adaptive biochemical reactions on early terms of inflammation development. At the same time, perftoran essentially reduces exudate volume, however this effect is short-time upon single infusion of the preparation. PMID- 17402597 TI - [Gender differences in sensitivity to psychotropic drugs]. PMID- 17402598 TI - [The role of changes in the functional activity and the amount of p-glycoprotein in drug pharmacokinetics]. AB - Data on the structure of P-glycoprotein, the mechanisms of interaction with drugs, the general properties of substrates, the role in the distribution of drugs, and the modulation of their interactions are summarized. Being an ATP dependent transport agent in the reverse yield, P-glycoprotein participates in the intestinal secretion, limitation of the permeability of histohematic barriers, and the renal and bile excretion of exogenous substrates, thus protecting the organism against xenobiotics. The interactions of drugs on the level of P-glycoprotein either leads to a decrease in these effects, when this transporter is activated by one of the drugs, or causes undesired reactions if P glycoprotein is inhibited. Poly- or mononucleotide polymorphism of MDRI gene encoding P-glycoprotein can lead to a change in the pharmacokinetics of related substrates. Pharmacotherapy must take into account both the substrate specificity of drugs with respect to P-glycoprotein and the individual features of patients, in particular, race and sex (allele homozygotes versus heterozygotes). PMID- 17402599 TI - An evidence-based approach to nipple feeding in a level III NICU: nurse autonomy, developmental care, and teamwork. AB - Historically, nipple feeding in the NICU has been initiated and advanced by physician order based on nursing observation and suggestions. The growing evidence base regarding nipple feeding readiness and progression, contingent caregiving, and nurse autonomy in decision making prompted our Level III NICU to reconsider our process. A literature review and discussions resulted in new protocols for initiating and advancing nipple feedings for three distinct groups: healthy preterm infants, preterm infants with complicated medical courses, and sick term or posttcrm infants. Through incorporating contingent earegiving, greater autonomy for the bedside nurse, developmental support, and collaboration, we expect to improve outcomes, comnuinication, and problem solving. PMID- 17402600 TI - Newborn screening: a literature review. AB - Newborn screening is the largest genetic testing effort for newborns in the U.S. Its purpose is to identify newborns who are at risk for metabolic, endocrine, or hematologic disorders. A review of the literature was conducted to determine the benefits of newborn screening; specimen collection timing and handling; ethical considerations of screening; as well as current practices regarding consent, notification of results, and follow-up procedures. The use of tandem mass spectrometry for expanded newborn screening and postmortem diagnosis of unexplained infant death was also reviewed. This article is intended to educate health care providers in the areas of controversy that surround the U.S. newborn screening program, with the hope of encouraging further research in this mportant area of newborn care. PMID- 17402601 TI - The chasm in neonatal outcomes in relation to time of birth in Lebanon. AB - PURPOSE: The purpose of this study was to investigate the relationship between the time of birth and the mortality and morbidity of infants admitted to neonatal intensive care units. DESIGN: This prospective, cohort study examined the records of women and infants admitted to the NICUs of four hospitals in Beirut, Lebanon, between July 1, 2002, and June 30, 2003. The hospitals selected were university affiliated and had a large number of deliveries (5,152 total for the year 2002 2003). MAIN OUTCOME VARIABLES: Neonatal mortality and morbidity for infants admitted to the NICU were evaluated in relation to time of birth. RESULTS: For the whole sample, mortality was higher for infants born during the night shift than for those born during the day shift. Mortality, morbidity, and brain asphyxia rates were also higher for infants born during the night shift and admitted to the NICU. Maternal risk factors and delivery complications were nor consistently higher on the night shift. PMID- 17402602 TI - Retained central venous lines in the newborn: report of one case and systematic review of the literature. AB - The use of percutaneously inserted central catheters (PICCs) is an established practice in most NICUs. With the widespread use of these catheters, an increasing number of PICC complications has been reported in neonates. We present one case of a PICC retained and tethered in the vein in a very low birth weight infant, as well as a systematic review of PICC retention cases. Ten previous cases of PICC retention in neonates were found. Among those cases, the most common associated factors were coagulase-negative staphylococcus catheter colonization/bactcremia and long duration of catheterization. Occlusion was not a usefiul sign for predicting catheter retention. Once retention was established, heparin or urokinase instillations were ineffective. Although surgical exploration was sometimes required, firm, continuous traction applied to the catheter over several hours or intermittent, moderately strong pulling maneuvers were successful in 44.4 percent of the cases in which they were performed. Traction did not show major side effects. Our case provides additional support for use of traction as the initial approach in the management of this rare complication. PMID- 17402603 TI - Building a case for neonatal palliative care. AB - The concept of palliative care, well recognized in the adult population, has not been fully implemented in the pediatric population. Yet there is an urgent need to define and provide excellence in end-of-life care for infants and their families. Beneficent end-of-life care for the dying neonate includes efforts directed at comfort care, assistance with end-of-life decision making, and bereavement support. Through research and education, the challenges to implementing neonatal palliative care programs can be overcome. This article describes the components of neonatal palliative care, identifies the challenges associated wiith the implementation of such programs, and proposes strategies for addressing these challenges. PMID- 17402604 TI - Potassum and sodium homeostasis in the neonate. PMID- 17402605 TI - Full disclosure when bad things happen. AB - Full disclosure of errors to family members of patients in the NICU starts the healing process and helps to restore confidence and faith in caregivers. By facilitating and encouraging full disclosure, managers help initiate the healing process for all involved and can better ensure regulatory compliance. PMID- 17402606 TI - Surgical retrieval of a transected umbilical artery catheter. PMID- 17402607 TI - A novel method for the detection of porcine circovirus type 2 replicative double stranded viral DNA and nonreplicative single stranded viral DNA in tissue sections. AB - Porcine circovirus type 2 (PCV2), an economically important pathogen of swine, is the necessary cause of post weaning multisystemic wasting disease (PMWS); PCV2 infection is associated with porcine dermatitis and nephritis syndrome (PDNS). Current immunohistochemical (IHC) methodologies identify PCV2 antigens but are not capable of differentiating replicating virus from nonreplicating virion particles in tissue sections. In this paper, a combination of IHC using commercial monoclonal antibodies specific for single stranded (ss) and double stranded (ds) DNA and PCV2 specific in situ hybridization (ISH) was used to show the specificity of the former for PCV2 DNA in tissue sections from PCV2-infected gnotobiotic pigs. Cold-ethanol-fixed tissue sections were superior to formalin fixed tissues for detection of PCV2 DNA, presumably due to the lack of protein cross-linking in the latter. These data demonstrate that conventional IHC detects PCV2 DNA forms in experimentally infected PCV2-positive gnotobiotic porcine tissue sections that are minimally compromised by either formalin fixation or the hybridization conditions needed for ISH. PMID- 17402608 TI - Identification and characterization of two bovine spongiform encephalopathy cases diagnosed in the United States. AB - Bovine spongiform encephalopathy (BSE) is a transmissible spongiform encephalopathy of cattle, first detected in 1986 in the United Kingdom and subsequently in other countries. It is the most likely cause of variant Creutzfeldt-Jakob disease (vCJD) in humans, but the origin of BSE has not been elucidated so far. This report describes the identification and characterization of two cases of BSE diagnosed in the United States. Case 1 (December 2003) exhibited spongiform changes in the obex area of the brainstem and the presence of the abnormal form of the prion protein, PrP(Sc), in the same brain area, by immunohistochemistry (IHC) and Western blot analysis. Initial suspect diagnosis of BSE for case 2 (November 2004) was made by a rapid ELISA-based BSE test. Case 2 did not exhibit unambiguous spongiform changes in the obex area, but PrP(Sc) was detected by IHC and enrichment Western blot analysis in the obex. Using Western blot analysis, PrP(Sc) from case 1 showed molecular features similar to typical BSE isolates, whereas PrP(Sc) from case 2 revealed an unusual molecular PrP(Sc) pattern: molecular mass of the unglycosylated and monoglycosylated isoform was higher than that of typical BSE isolates and case 2 was strongly labeled with antibody P4, which is consistent with a higher molecular mass. Sequencing of the prion protein gene of both BSE-positive animals revealed that the sequences of both animals were within [corrected] the range of the prion protein gene sequence diversity previously reported for cattle. PMID- 17402609 TI - A latex agglutination test for the rapid detection of avian influenza virus subtype H5N1 and its clinical application. AB - A rapid and simple latex agglutination test (LAT) for the detection of avian influenza virus (AIV) subtype H5N1 in chicken allantoic fluids, tracheal swabs, and tissues was developed. Monoclonal antibodies against the hemagglutinin glycoprotein of H5N1 were covalently coupled onto the surface of carboxylated latex bead using a water-soluble carbodiimide to obtain sensitized latex particles (SLP). These SLPs strongly agglutinated in the presence of allantoic fluid containing H5N1, but not fluids containing other AIV sub-types such as HIN1, H3N2, H4N6, and H9N2. Using this LAT, the virus was detectable in tracheal swabs 24 hours to 30 days after inoculating chickens with H5N1, with detection rates ranging from 45.5 to 79.2%. Much higher rates of detection were obtained from tissues collected postmortem from H5N1 experimentally infected chickens; lung tissue yielded the highest detection rate (96.7%), followed by kidney, spleen, brain, and liver tissues (90%). Lower detection rates were achieved with heart (41.7%) and cloacal tissues (26.8%). When the LAT was compared with other detection methods, the agreement with the viral isolation, H5 antigen immunochromatographic test,and H5 real-time RT-PCR test was 93.97, 95.18, and 87.95%, respectively. The test was highly specific for H5N1 in chickens and water fowls and had sensitivity comparable to other diagnostic tests evaluated. PMID- 17402610 TI - Detection of antimicrobial resistance genes of pathogenic Salmonella from swine with DNA microarray. AB - A glass-based microarray was developed to detect 11 antimicrobial resistance genes that confer resistance to aminoglycosides, tetracyclines, sulfonamides, and chloramphenicols. The target genes for microarray were generated from Salmonella isolates by PCR and confirmed by sequencing. The specificity of the microarray was tested using 11 positive DNA probes. The sensitivity was tested with tetA gene and Salmonella isolates. Using detection threshold of signal-to-noise ratio (S/N) > or = 1.5 or median pixel intensity > or =1000, antimicrobial resistance genes carried by 30 Salmonella isolates were detected. Common genes included sul I(76.7%, 23/30), aph(3')-IIa (60%, 18/30), tetC (60%, 18/30), cat] (43.3%, 13/30), tetA (40%, 12/30) and aadA1 (36.7%, 11/30), and the results were confirmed to be correct by PCR. PMID- 17402611 TI - Aflatoxicosis in nine dogs after exposure to contaminated commercial dog food. AB - The purpose of this study was to characterize light and electron microscopic findings from 9 dogs that had consumed aflatoxin-contaminated commercial dog food from recalled batches. Four dogs died and 5 were euthanized after signs of liver failure. Analysis of feed and liver samples confirmed exposure to aflatoxin. Of the 9 dogs, 8 had classic signs of liver failure, and 1 had signs of liver failure. Enlarged, pale yellow livers were seen macroscopically at necropsy in the dogs with subacute hepatopathy, and cirrhosis was noted in the dog with chronic hepatopathy. Histopathologic findings included hepatic lipidosis, portal fibroplasia, and biliary hyperplasia, which supported a diagnosis of subacute toxic hepatopathy in the 8 symptomatic animals. Marked lobular atrophy, bridging portal fibrosis, and regenerative hepatocellular nodules characterized the dog with chronic hepatopathy. Electron microscopy revealed marked hepatocellular lipid vacuolation and early fibroplasia in the dogs with acute hepatopathy and marked fibrosis and regeneration in the dog with chronic hepatopathy. Analysis of feed for aflatoxin consistently revealed high levels of aflatoxin B1 (range of 223-579 ppb), and hepatic tissue contained elevated levels of aflatoxin B1 metabolite M1 (0.6-4.4 ppb). Although dogs are not commonly affected by aflatoxicosis, they are highly susceptible and can present with classic signs of acute or chronic hepatopathy. Characteristic gross, histologic, and electron microscopic changes help pathologists determine a presumptive toxic insult. Detecting aflatoxins or their metabolites in feed or liver specimens can help confirm the diagnosis of aflatoxicosis. PMID- 17402613 TI - Toward a multiplexed serotyping immunoassay for foot-and-mouth disease virus. AB - Initial results demonstrating the feasibility of a multiplexed liquid array immunoassay for foot-and-mouth disease viral antigen detection and simultaneous serotype differentiation are presented. Serotype-specific antibodies from rabbit and guinea pig hyperimmunesera were isolated and prepared for use in a multiplexed, bead-based assay. The performance of all of the available antibodies as both capture and detector reagents was evaluated in the multiplexed system to establish a combination exhibiting the highest homotypic responses and lowest heterotypic reactions. The multiplexed assay was evaluated against inactivated cell culture supernatant samples of the same subtype as the virus used to raise the capture and detector antibodies. Distinct serotype differentiation was observed, except in the case of serotype SAT1. Subsequently, cell culture supernatant samples from a larger pool of viral subtypes were analyzed. Distinct serotype differentiation was obtained when analyzing cell culture supernatant samples from viral serotypes C, Asia, and SAT3, irrespective of the subtype. However, limitations of the current antibody pairs were realized in some inconclusive results obtained when analyzing samples from a broader range of O, A, and SAT2 subtypes. The results obtained in this initial study will be used to further optimize the assay using polyvalent or monoclonal antibodies and move toward the analysis of clinical samples. PMID- 17402612 TI - Serological prevalence of canine respiratory coronavirus in southern Italy and epidemiological relationship with canine enteric coronavirus. AB - Canine respiratory coronavirus (CRCoV) has been detected in dogs suffering from respiratory disease and is thought to be involved in canine infectious respiratory disease (CIRD) complex. Canine enteric coronavirus (CECoV) is a widespread pathogen of dogs, responsible for mild to severe diarrhea in pups. The purpose of this study was to establish the seroprevalence of CRCoV in Italy and its relationship to CECoV type II seroprevalence. The age and year of sample collection from seropositive dogs was also assessed. Of adult domestic dogs, 23.3% had antibodies to CRCoV, compared with 86.1% with antibodies to CECoV. Amongst a population of kenneled pups, 4.0% had antibodies to CRCoV, and 97.0% had antibodies to CECoV. PMID- 17402614 TI - Clostridium perfringens type A toxin production in 3 commonly used culture media. AB - In vitro toxin production is an important tool not only for diagnostic purposes but also for the study of pathogenesis of Clostridium perfringens infections. The present study was carried out to compare the level of toxin production by several strains of C. perfringens type A, isolated from the intestine of animals, when cultured in 3 different conventional culture media. Six strains of C. perfringens type A isolated from the small intestine of healthy sheep were cultured in commercial cooked meat medium (CMM), brain heart infusion (BHI), and tryptone glucose yeast (TGY). Intravenous lethality in mice and phospholipase C (PLC) activity were measured in filtered culture supernatants. Lethality of culture supernatants was highest for all isolates when grown in BHI, followed by CMM. No supernatants from any isolates grown in TGY produced lethality in mice. Phospholipase C activity was highest when the isolates were grown in BHI and CMM and significantly lower when grown in TGY. PMID- 17402615 TI - Impact of Corynebacterium pseudotuberculosis infection on serologic surveillance for Johne's disease in goats. AB - False-positive results on serologic assays for Mycobacterium avium subsp. paratuberculosis (MAP) are believed to occur due to cross-reacting antibody produced by Corynebacterium pseudotuberculosis (C. pstb) infection in goats. This issue of compromised specificity was evaluated by testing 771 adult goats from 10 Midwestern goat herds in 2004. Assays for MAP infection included radiometric fecal culture and 2 enzyme-linked immunosorbent assays (ELISAs); ELISA-positive samples were tested by agar gel immunodiffusion (AGID). A synergistic hemolysin inhibition assay (SHI) was used to detect C. pstb antibody. Four infection status categories were evaluated. Category 1 goats (free of both MAP and C. pstb infection) tested negative on all MAP fecal cultures and SHI tests. Five of 181 goats were positive in both ELISAs, and 2 more were positive in ELISA-1 only. For Category 2 (MAP infected; no C. pstb infection), all animals were SHI negative. Six goats were fecal culture positive and strongly positive in both ELISAs; 2 more goats were positive only in ELISA-1. For Category 3 (C. pstb infected or vaccinated; no history of MAP infection), all fecal cultures were negative and 91% were SHI test-positive. In this population, only 2 goats were positive in both MAP ELISAs, while 84 additional goats were test-positive only on ELISA-1. In the absence of C. pstb infection, both ELISAs performed comparably, but when C. pstb infection was present the performance of ELISA-1 was significantly perturbed. Use of the ELISA-2 for goats is an effective and efficient method for Johne's disease surveillance in any goat herd. PMID- 17402616 TI - Toxoplasma gondii in an African crested porcupine (Hystrix cristata). AB - An adult female crested porcupine (Hystrix cristata) was evaluated for acute onset of neurologic signs including head tilt, circling, and ataxia. She was found dead in her holding area 2 days after initially exhibiting clinical signs. Necropsy was unremarkable. Histopathology of brain tissue revealed the presence of protozoal cysts associated with inflammation as the underlying cause of clinical signs and death. Immunohistochemical staining of brain tissue for Toxoplasma gondii was strongly positive. PCR on fresh brain confirmed T. gondii as the causative organism. An adult male in the same enclosure has demonstrated similar neurologic signs over the past 3 years and has failed to respond to various medical treatments. Clinical disease associated with T. gondii has not been previously reported in this porcupine species or any other Old World porcupines, although there are several reports of clinical toxoplasmosis involving New World porcupine species. PMID- 17402617 TI - Unilateral orchitis and epididymitis caused by Salmonella enterica subspecies diarizonae infection in a ram. AB - A case of unilateral suppurative epididymo-orchitis associated with Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) infection is described in a 2 year-old ram. Gross lesions were characterized by severe enlargement of the scrotal contents, fibrous adhesions between testicular layers, coexistence of epididymal abscesses and foci of fibrinous exudate, and testicular atrophy. Microscopically, testicular and epididymal microabscesses and diffuse inflammatory infiltrates with abundant macrophages containing short Gram-negative rods were observed. Superimposed on the chronic lesions were fibrin deposits with clusters of neutrophils, as well as walled-off granulation tissue. Bacterial colonies were also identified in thrombosed spermatic cord vessels, scrotal lymph nodes, lung, and liver. S. enterica subsp. diarizonae serovar 61:k:1,5,(7) was isolated from the affected testis. To the authors' knowledge, this is the first report of infection of the testis and epididymis by Salmonella in rams. This organism must be taken into account in the differential diagnosis of ovine genital infections. PMID- 17402618 TI - Muscular pseudohypertrophy (steatosis) in a bovine fetus. AB - Muscular pseudohypertrophy was diagnosed in the cervical musculature of a full term crossbred Simmental fetus delivered by fetotomy. Only head and cervical regions were submitted for pathologic examination; the rest of the fetal body was reportedly normal. The neck musculature of the fetus was markedly deformed by 23 cm and 18 cm in diameter, firm, spherical masses that consisted of enlarged and pale left splenius and right serratus ventralis cervicis muscle, respectively, covered by intact skin. Additionally, lipomatous masses were present within the cervical vertebral canal, compressing the spinal cord. Microscopically, the prominent muscular enlargement was due to massive adipose and fibrous connective tissue replacement of atrophic muscle. Focal myelodysplasia and astrocytosis affecting the grey matter was detected in the mid-cervical region of the spinal cord, accompanied by degeneration in the ascending and descending tracts of the remaining cord segments. Abnormal spinal cord development as a result of severe spinal cord compression by the lipomatous masses within the spinal canal leading to replacement of muscle by fat and fibrous tissue was considered to be the cause of the muscular malformation in this fetus. PMID- 17402619 TI - Prevalence of neoplasia in llamas and alpacas (Oregon State University, 2001 2006). AB - Prevalence and type of neoplastic disease were determined in 551 camelid submissions (368 alpacas [Lama pacos], 180 llamas [Lama glama], and 3 cases in which species was not identified) over a 5-year period. Forty neoplasms were identified in 38 animals (6.9%). Prevalence of neoplasia in llamas was higher (11%) than in alpacas (4.9%). Mean age of camelids with neoplasia was 9.42 +/- 4.9 years. Mean age of alpacas with neoplasia (5.48 +/- 3.7 years) was significantly less than of llamas with neoplasia (12.53 +/- 3.2 years; P < 0.001). Cutaneous and mucocutaneous fibroma/fibropapilloma was most common (10 animals), followed by cutaneous and mucocutaneous squamous cell carcinoma (6 animals), disseminated lymphoma (5 animals), and fibrosarcoma (4 animals). Four of 5 animals with lymphoma were alpacas, aged 0.21 to 4 years. Lymphoma occurred in 1 aged llama (15 years). Disseminated carcinoma and adenocarcinoma occurred in 4 llamas and 2 alpacas, and included biliary (2), gastrointestinal (2), mammary gland (1), and unknown (1) origin. Mean age of camelids with any type of carcinoma or adenocarcinoma (12.36 +/- 2.8 years) was significantly greater than that of camelids with lymphoma (4.24 +/- 6.2 years; P = 0.02). Results indicate that neoplasia is relatively common in camelids and that there are differences between llamas and alpacas as regards prevalence of neoplasia, tumor types, and age at diagnosis. PMID- 17402620 TI - Vertebral polyostotic lymphoma in a young dog. AB - An unusual clinical presentation of lymphoma with vertebral involvement in a dog is reported. A 20-month-old intact female Golden Retriever presented with progressive paraparesis and anorexia. Complete blood count and serum biochemistry profile demonstrated pancytopenia and hypercalcemia. Ventral fusion of the lumbar vertebrae by new bony tissue deposition was evident on X-ray and CT scan. Fine needle aspiration revealed neoplastic lymphoid cells in lymph nodes and bone marrow. Histologically, vertebral bone and osteophytes, liver, bone marrow, kidney, and lymph nodes were diffusely infiltrated by neoplastic, lymphoid cells, with scant cytoplasm and round hyperchromatic nuclei. Polyostotic and medullary T cell lymphoma with spondylosis was diagnosed. Lymphoma mainly affecting bone is uncommon in the dog. The present case differs from previously described polyostotic lymphomas in clinical signs of the disease, mainly attributable to spondylarthrosis. In addition, lymphomatous proliferation was associated with osteoproductive lesions of the vertebrae. PMID- 17402621 TI - Kernicterus in a neonatal foal. AB - A 5-day-old Thoroughbred foal was submitted to the necropsy service at the University of Kentucky Livestock Disease Diagnostic Center. The foal had a clinical history of seizure activity and severe icterus. A complete blood count and serum chemistry analysis indicated that the foal was anemic (hematocrit, 16%), hyperbilirubinemic (45 mg/dl), and hypoglycemic. At necropsy, all tissues were discolored various shades of yellow. Microscopically, there was degeneration and necrosis of cerebral neurons and cerebellar Purkinje cells; severe hepatocellular degeneration and necrosis; and deposition of amorphous golden yellow material in the cerebellar granular cell layer, pulmonary alveoli, renal tubular epithelium, splenic trabecula, and the lamina propria of the small and large intestine. The golden-yellow material in the brain, lung, spleen, and small intestine was identified as bilirubin by histochemistry. Based on the macroscopic and microscopic findings, a diagnosis of kernicterus (bilirubin encephalopathy) was made. This report describes a rare case of equine neonatal kernicterus. PMID- 17402622 TI - Arsenic and metaldehyde toxicosis in a beef herd. AB - Over a 12-day period, 13 animals in a herd of 110 beef cattle developed ataxia with profound muscle fasciculations progressing to recumbency. Twelve animals (5 adults and 7 calves from 8-10 months of age) died, and 1 cow was euthanized. Hemorrhagic diarrhea occurred in some, but not all, animals. The onset of clinical signs was at least 12 hours after the cattle had gained access to contents of old buildings used for storage, and the majority of deaths occurred within 24 to 48 hours after the onset of clinical signs. Approximately 9 kg of unidentified pellets were found strewn in the barn area where the cattle had been. Autolysis considered more severe than expected for the postmortem interval, suggestive of high body temperature before death, and congestion of body tissues were the only significant findings detected in the cow that was euthanized and submitted for necropsy examination. The clinical history and lack of postmortem lesions were most consistent with toxicity. A toxic level of arsenic (6.18 ppm) was detected in the kidney, and metaldehyde was detected in the liver. The pellets were analyzed and found to contain both arsenic and metaldehyde, consistent with a discontinued molluscicidal product. PMID- 17402623 TI - Fatal ricin toxicosis in a puppy confirmed by liquid chromatography/mass spectrometry when using ricinine as a marker. AB - Ricin, a lectin from the castor bean plant (Ricinis communis), is considered one of the most potent plant toxins. Ingestion of masticated seeds results in high morbidity, with vomiting and watery to hemorrhagic diarrhea. The prognosis varies with the number of seeds ingested, the degree of mastication, individual susceptibility, and the delay in treatment. Low mortality restricts assessment of histologic lesions, and the literature on toxicologic analysis for ricin is limited. This report describes a fatal case of castor bean ingestion in a 12-week old Mastiff puppy, with confirmation of ricin exposure through detection of the biomarker ricinine by liquid chromatography/mass spectrometry (LC/MS). Despite supportive therapy, the puppy died several hours after presentation for acute vomiting, diarrhea, and lethargy. At necropsy, a segment of jejunum and mesenteric lymph nodes were congested. When the owner reported the presence of castor beans in the dog's feces, selected formalin-fixed and unfixed tissues were submitted for diagnostic evaluation. Histopathologic findings included superficial necrotizing enteritis of the jejunum and occasional, random foci of coagulative necrosis in the liver. The alkaloid ricinine was detected in gastric content by using a newly developed LC/MS method. This confirmation of exposure is important in the diagnosis of ricin toxicosis, because ingestion of castor beans is not always fatal, histologic lesions are nonspecific, and the degree of mastication can influence the effective dose of ricin. PMID- 17402624 TI - Taxus sp. intoxication in three Francois' langurs (Trachypithecus francoisi). AB - Three Francois' langurs (Trachypithecus francoisi) were found dead, without previous clinical signs, over a 48-hour period at a zoological institution after transfer to a new exhibit. A hybrid yew shrub (Taxus baccata X T. cuspidata) was found in close proximity to the exhibit perimeter fence. Despite clinical suspicion of yew intoxication, thin-layer chromatography performed on gastric contents was negative. However, microscopic examination of gastric contents revealed multiple yew fragments, and taxine alkaloids were detected by gas chromatography and mass spectrometry of the gastric contents to confirm yew intoxication. Acute death of the animals prevented treatment. The fourth langur in the collection survived, most likely because of its low rank in the troop's hierarchy, with a suspected small amount or none of the plant ingested. To the authors' knowledge, this case report is the first yew intoxication documented in a nonhuman primate species. Taxus spp. intoxication is an often fatal condition reported in domestic animals and humans. In comparison with these species, mortality appeared delayed in the Francois' langurs, most likely because of their unique gastrointestinal anatomy, with both foregut and colonic fermentation. Plant intoxication should be a differential diagnosis when multiple acute deaths are observed after recent introduction to a new enclosure. PMID- 17402625 TI - Oral cancer: Part II--risk assessment and screening protocol. PMID- 17402626 TI - HIV at twenty five and counting. PMID- 17402627 TI - A polychromatic composite layering approach for solving a complex Class IV/direct veneer/diastema combination: Part II. AB - Restoration of a Class IV defect in the aesthetic zone in combination with a direct resin veneer can be a considerable challenge for clinicians, especially when striving to emulate the natural dentition. Shade-matching is a particular concern in achieving a natural-looking aesthetic result. Value enamels can also be used over selective areas of a Class IV restoration as needed to modify the perception of the chroma of underlying layers of artificial dentin and body enamel. This second and concluding part provides a discussion of how an advanced polychromatic composite layering technique can be used to meet the patient's aesthetic expectations. PMID- 17402628 TI - Utilization of an Er,Cr:YSGG laser for the removal of all-ceramic restorations. PMID- 17402629 TI - The impression process: Part III--technique. PMID- 17402630 TI - Conservative restorative treatment using a single-visit, all-ceramic CAD/CAM system. AB - Computer-aided design/computer-aided manufacturing (CAD/CAM) continues to radically change the way in which the dental team plans, prepares, and fabricates a patient's restoration. This advancing technology offers the clinician the ability to scan the patient's failing dentition and then designs a long-lasting, reliable restoration based on this data. CAD/CAM systems also permit efficient, single-visit placement of the restoration while preserving much of the natural tooth structure. This article discusses how a chairside CAD/CAM system can be used to provide such a restoration in the posterior region in a single-visit. PMID- 17402631 TI - High quality and convenience with digital radiography in pediatric care. PMID- 17402632 TI - Intracoronal bleaching of nonvital discolored mandibular incisors. AB - Intracoronal bleaching of pulpless discolored mandibular incisors is a valuable treatment modality currently disregarded by many clinicians because of the potentially disastrous consequence of cervical resorption. A patient administered, intracoronal carbamide peroxide bleaching technique is described. This modified, walking bleaching method minimizes the risks because treatment time is reduced to days, as opposed to weeks with the original walking bleaching protocol, the concentration of the hydrogen peroxide is markedly reduced, and residual hydrogen peroxide is completely eliminated with the use of catalase prior to the definitive restoration. PMID- 17402633 TI - Soft tissue recession around implants: is it still unavoidable?--Part I. AB - When treatment with dental implants is indicated, an accurate diagnosis must be made to evaluate the clinical parameters and determine the optimal time for immediate or delayed (ie, early or late) implant placement and loading following tooth extraction. It is also important to identify complications and their implications on the aesthetic outcome. This article explains the behavior of the hard and soft tissue around the implant, evaluates the timing of implant placement after extraction, and reviews various parameters that influence tissue marginal remodeling. PMID- 17402634 TI - The interface between biomaterials science and biotechnology. AB - Biotechnology has evolved over the years, moving on from crop protection to drug discovery. Similarly, biomaterials science has moved forward from implantable medical device technology to drug and gene delivery and tissue engineering. It was perhaps inevitable that they should eventually meet. PMID- 17402635 TI - The use of power beams in surface modification. AB - This review of surface modification technologies focuses on the use of power beams, that is, lasers and electron beams, to modify polymer surfaces. A novel electron-beam materials processing technique is also described that offers scope for generating new types of surface functionality on metals used to construct stents and implants. PMID- 17402636 TI - Adhesive solutions for medical electronic devices. AB - Today's adhesives provide more than just bonding. They play an important role in solving problems associated with thermal and electrical conductivity, shielding, grounding, sealing and vibration damping. PMID- 17402637 TI - Modifying surfaces and interfaces for improved biomaterial performance. AB - Greater understanding of the fundamental processes that drive a body's response to an implant material is leading to new approaches in biomaterial surface engineering. Some recent work is reported. The discussion looks, respectively, at the important roles of the surface water layer and microscopic molecular domains, the new emphasis on conducting polymers and future developments. PMID- 17402638 TI - Improving the surface friction of silicone elastomer parts. AB - The relatively high coefficient of friction of silicone elastomers can limit the range of applications of these materials. A liquid silicone rubber top coat can be used to improve surface friction of silicone elastomer parts. It can be applied using conventional coating methods. The performance benefits it offers and potential product applications are described. PMID- 17402639 TI - A holistic approach to airflow measurements in clean rooms. AB - How and when to accurately measure airflow are important issues in achieving optimum efficiency in a clean room. This article describes the equipment to use, the pros and cons of those methods and adds to the current debate over frequency of measuring. PMID- 17402640 TI - Design reuse and medical devices. AB - Design reuse is an important factor in consumer electronics, where removing the burden of designing basic functions for each product can realise critical savings in time to market and delivered cost. But designers of medical devices must be more circumspect and selective to gain tangible advantages from design reuse. PMID- 17402641 TI - Insurance coverage for European device clinical studies. AB - The requirements for insurance coverage of subjects participating in European medical device clinical studies, and the cost of this coverage, vary significantly throughout Europe. This article discusses this variability and the importance of identifying insurance coverage needs in the early stages of planning European clinical studies. PMID- 17402642 TI - Sample preparation in lab-on-a-chip systems. AB - Fully integrated lab-on-chip systems are relatively rare, and no current system can claim to be able to perform all the roles of a laboratory. This article outlines ways of combining specific single modules to form a fully integrated lab on-a-chip system for deoxyribonucleic acid analysis. PMID- 17402643 TI - Using manufacturing marks to identify counterfeits. AB - Manufactured objects can now be identified by individual characterisitics, just as humans can be identified by their irises or fingerprints. Using microscopic imperfections generated by the manufacturing process and the raw materials used to produce an object, digital imaging technologies can generate a unique "imprint." Therefore, it is possible to identify and to trace a medical device during its entire life. PMID- 17402644 TI - Fostering innovation and collaboration. AB - The Health Technologies Knowledge Transfer Network is taking active steps to foster innovation and collaboration. Users of its internet portal can access a range of valuable tools to aid this process, including free virtual meeting facilities. PMID- 17402645 TI - [No response to antihypertensive therapy: consider non-adherence]. AB - In 3 patients, 2 men aged 62 and 43 years, respectively, and 1 woman aged 53 years, the medication prescribed to reduce blood pressure was insufficiently effective. Drug adherence was questioned. The first patient was afraid of the side effects mentioned in the medication information leaflet. The second patient had insufficient plasma levels of the medication but would not discuss the subject. The blood pressure of the third patient was uncontrolled despite treatment with three antihypertensive drugs. During a short hospital stay, her blood pressure responded favourably to treatment with only one of the three drugs. Subsequent repeated instruction did not improve her situation. Drug adherence is an important issue in daily clinical practice. In patients with asymptomatic conditions like hypertension, adherence is expected to be poor and worsen over time. Adherence is very difficult to measure. Risk factors for poor adherence include complex medication schedules, multiple dosing times, depression and real or suspected side effects. Measures to improve drug adherence include simplifying the medication schedule, discussing the schedule during check-ups, using automated blood pressure measuring devices at home, maintaining e-mail contact with the patient and involving a specialised nurse. Increased awareness of poor adherence is an important step toward improving hypertension treatment. PMID- 17402646 TI - [Anaphylaxis: facts and fallacies]. AB - The diagnosis and management of anaphylaxis is surrounded by many misunderstandings. The diagnosis may be made ifa patient develops respiratory or circulatory symptoms, in addition to skin symptoms, after exposure to a possibly relevant allergen. The most common causes ofanaphylaxis are food allergens, such as peanut and tree nuts, insect stings, and drugs, in particular antibiotics. Many patients with peanut or tree nut allergy show only mild allergic symptoms; only a minority develop anaphylaxis upon exposure. A large local reaction to an insect sting does not constitute insect sting allergy and does not increase the risk of anaphylaxis when stung again. Intramuscular epinephrine is the drug of choice in the treatment of anaphylaxis; antihistaminic agents and corticosteroids are supportive therapeutic agents which should only be considered after epinephrine has been administered. Prevention of recurrence ofanaphylaxis comprises identification and avoidance of the causative allergen, and administration of epinephrine autoinjector when the patient develops suggestive signs of recurrence. When prescribing an epinephrine autoinjector, the patient and all caregivers should be instructed carefully in its correct use; referral to an allergist is recommended. PMID- 17402647 TI - [The development of memory clinics in The Netherlands]. AB - The development of memory clinics in the Netherlands. - Memory clinics (MCs) are multidisciplinary teams involved in the early diagnosis and treatment of people with dementia. Between 1998 and 2004 the number of MCs in the Netherlands increased from 13 to 40. This type of clinic is increasingly becoming part of standard care for people with early dementia and other cognitive disorders. A growing number ofMCs are collaborating structurally with local mental health care service providers. This is a positive development, in which hospital-based diagnosis and treatment are increasingly being integrated with long-term care to reach patient centred disease management. PMID- 17402648 TI - [Hyponatraenmia during a long-distance run: due to excessive fluid intake]. AB - Hyponatraemia during a long-distance run, such as a marathon, is usually mild and asymptomatic. However, in runners who present with symptoms at a healthcare station, the diagnosis of hyponatraemia can have significant consequences. The complaints are usually aspecific, but in more severe hyponatraemia, signs of cerebral dysfunction due to incipient to severe cerebral oedema predominate. The most important aetiological factor is an excessive electrolyte-free fluid intake. The most important risk factors for the development of hyponatraemia are: a long duration of the run, female gender, recent use of NSAIDs, lower body weight and environmental factors like high temperature and high humidity. In the presence of symptoms, rapid correction must be started by administration ofhypertonic saline, sometimes in combination with a loop diuretic in cases of eu- and hypervolaemia. Isotonic saline must be added in case of dehydration. Prevention consists primarily of advice to moderate the fluid intake. PMID- 17402649 TI - [TNF-blocking drugs and infection; recommendations for daily practice]. AB - Anti-tumour necrosis factor (TNF) therapy is associated with an increased risk of infection. There are sparse data and no evidence-based guidelines on how to deal with this problem in daily practice. However, recommendations can be made based on theoretical considerations and by extrapolating from recommendations for other types ofimmunodeficiency. Before starting anti-TNF therapy, screening for tuberculosis and other possible infections is indicated. During therapy, alertness is required to the increased risk of infection, infections with a more serious clinical course or unusual manifestations and opportunistic infections. Flu vaccination during anti-TNF therapy is indicated. Travel vaccinations with live microbial inocula should not be given. PMID- 17402650 TI - [From the Cochrane Library: Marginally higher chance of cure by antibiotic treatment in acute bacterial conjunctivitis]. AB - Acute bacterial conjunctivitis is one of the most frequently encountered ocular disorders in primary care. It is frequently self-limiting, and the widespread use of broad-spectrum antibiotics has led to concerns regarding antibiotic resistance. Therefore, a Cochrane systematic review of 5 randomised clinical trials that compared antibiotic treatment with placebo in patients with acute bacterial conjunctivitis was recently updated. The chances of clinical and microbiological benefits of topical antibiotics were small but statistically significantly higher compared with placebo in the early stage (2 to 5 days after the first day of the intervention) and late stage (6 to 10 days after the first day of the intervention). The risk of adverse events in patients treated with placebo appeared to be low. Future trials should assess the cost-effectiveness of antibiotic treatment for acute bacterial conjunctivitis. PMID- 17402651 TI - [Diagnostic image (314). A man with a headache and haemorrhagic nasal discharge]. AB - A 56-year-old man presented with an oronasal fistula in the palatum durum due to cocaine sniffing. PMID- 17402652 TI - [Serious haemorrhage after conventional (adeno)tonsillectomy: rare and most often on the day of the procedure]. AB - OBJECTIVE: To analyse the incidence and timing of postoperative haemorrhage that requires re-operation following tonsillectomy or adenotonsillectomy, and to formulate a recommendation regarding the optimal duration of postoperative clinical observation. DESIGN: Retrospective. METHOD: Data were collected on the incidence of haemorrhage that required exploratory surgery in all patients who underwent adenotomy (n=3508) or tonsillectomy or adenotonsillectomy (n=4909) in the period 1996-2002 at the Sint Lucas Andreas Hospital in Amsterdam, the Netherlands. The Sluder technique was used on an outpatient basis in 2439 of the 4909 patients who underwent tonsillectomy or adenotonsillectomy; these patients were aged <10 years. The remaining 2470 patients were aged > or =10 years and underwent conventional dissection followed by 24 hours of clinical observation before being discharged. RESULTS: Postoperative haemorrhage was recorded in 0 of the 3508 patients who underwent adenotomy, 12 (0.5%) of the 2439 patients aged <10 years who underwent tonsillectomy using the Sluder technique and 43 (1.7%) of the 2470 patient aged > or =10 years who underwent conventional dissection. Of the 43 cases of haemorrhage after conventional dissection, 31 (75%) occurred within 24 hours, including 2 cases that occurred between 12 and 24 hours (2/2470; 0.08%; 95% CI: 0.001-0.29). CONCLUSION: Reducing the postoperative observation period from 24 hours to 12 hours would have unfavourable consequences in 1 (95% CI: 0-3) per 1ooo patients operated. In regard to the risk of postoperative haemorrhage, outpatient tonsillectomy and adenotonsillectomy appear to be justified for patients aged 10 years or over, provided that surgery is performed in the morning and the patient is observed in a ward that is open until the evening. PMID- 17402653 TI - [Anaphylaxis in two children caused by peanut and nut allergies; recommendations for treatment]. AB - Two girls aged 12 and 7 years with asthma and peanut and nut allergy developed anaphylactic shock after ingestion of peanuts and nuts from an unreported source. They were both given intramuscular epinephrine. The 12-year-old girl was treated clinically for shock and after two days was discharged from hospital. The 7-year old girl died. Risk factors for life-threatening anaphylactic reactions are adolescent to young adult age, asthma, previous severe anaphylactic reactions to the food in question, previous reaction to small dose of the food in question and allergy to peanuts or tree nuts. A double-blind, placebo-controlled food challenge should be carried out to document the culprit food. The most important therapeutic intervention is the intramuscular administration of epinephrine. For patients with two or more risk factors the prescription of an epinephrine auto injector should be considered. PMID- 17402654 TI - [Two children with severe complications following incomplete removal of a percutaneous endoscopic gastrostomy (PEG) catheter]. AB - A 6-year-old boy with multiple severe disabilities was admitted with acute and progressive dyspnoea. A new percutaneous endoscopic gastrostomy (PEG) catheter had been placed 2 weeks earlier, during which the old catheter was cut and left in the stomach. Radiological assessment revealed pneumonia and a traumatic fistula between the oesophagus and the left main bronchus. Respiratory support was required. The patient recovered after oesophagoscopic removal of the remaining portion of the PEG catheter. A 7-year-old boy with multiple severe disabilities presented with an acutely reduced level of consciousness, vomiting and progressive dyspnoea. Chest x-ray revealed signs of aspiration pneumonia and, after respiratory problems worsened, a foreign object in the oesophagus. The foreign object was likely the remaining portion of a PEG catheter that was removed 12 months earlier. The patient was discharged in good condition a few days after oesophagoscopic removal of the remaining catheter. PEG is a commonly used method for enteral feeding in children. The Dutch guideline on enteral feeding in children indicates that endoscopic removal of the PEG catheter is often necessary. In daily practice, however, endoscopic removal is not always performed. To avoid serious complications, authors recommend endoscopic removal ofthe silicon disk when replacing or removing a PEG catheter in children aged less than 6 years and all children with mental retardation, prior laparotomy or constipation. Endoscopic removal of the disk should be considered in all other children if the disk is not passed in stool within 2 weeks and an x-ray shows that the disk is in the oesophagus, stomach or proximal intestine. PMID- 17402655 TI - [Proximal femur fractures in the elderly in The Netherlands during the period 1991-2004: incidence, mortality, length of hospital stay and an estimate of the care capacity needed in the future]. PMID- 17402656 TI - Advisory committees at FDA: the Hinchey Amendment and "conflict of interest" waivers. AB - In approving the Food and Drug Administration's (FDA) Fiscal Year 2007 budget, the House approved an amendment that would prevent the agency from using appropriated funds to waive certain conflicts of interest identified by members of its advisory committees. The amendment, introduced by Representative Hinchey and known as the Hinchey Amendment, provides that no funds may be used to: waive a conflict of interest under Section 505(n)(4) of the Federal Food, Drug, and Cosmetic Act (FDCA) for any voting member of an FDA advisory committee or panel; or make a certification under Section 208(b)(3) of Title 18 of the U.S. Code for any such voting member. This creates a problem, as ties to industry create the very expertise that FDA values in its outside advisors-under the Hinchey Amendment, these very ties would prevent them from serving as advisors to FDA during the drug approval process. The author opposes this change in the law and argues that the Hinchey Amendment would undermine congressional efforts in 1962 and 1989 to carefully balance the goals of attracting qualified experts and protecting agency decisionmaking. Further, the author argues, this change is unnecessary at FDA, because experts on FDA advisory committees divulge their connections to the industry, because the committees offer only advice and do not make agency decisions, and because the agency is under the watchful eye of Congress, the public, and public interest groups. The Article concludes that although FDA's advisory committee conflict-of-interest process can be improved, congressional action is unnecessary, and a change in the law through amendment to an appropriations bill that does not go through the ordinary legislative process (as an amendment to the FDCA or Title 18 would) is inappropriate. Instead, recommendations from organizations studying FDA practice, such as the OIG, GAO, and IOM, should be used to carefully and reflectively amend the process at the agency level, within the existing statutory framework. PMID- 17402657 TI - "No-hire" clauses in healthcare sector contracts: their use and enforceability. AB - In today's healthcare industry, many hospitals utilize outside agencies for both business and clinical functions. This Article acknowledges the prevalence of outsourcing contract labor in the healthcare arena and focuses on the restrictive provisions included in these employment contracts, particularly "no-hire" clauses. No-hire clauses are often included in contracts between healthcare providers and professional groups that provide clinical service employees to the provider, such as a medical practice group providing physicians to a hospital or an agency providing nurses to a nursing home. These clauses usually provide that the healthcare provider may not directly hire an employee provided by the professional group, nor may it contract with another professional group that later hires the employee. The purpose of a no-hire clause is two-fold: to protect the professional group's investment of time and moneyfor recruiting, training, and establishing the employee's clinical practice, and to give the professional group leverage to retain its employees. While noncompete clauses in employment contracts have traditionally been the subject of litigation, no-hire clauses raise distinct legal issues. Case law provides conflicting views as to the enforceability of these provisions. Some courts find no-hire clauses to be per se illegal restrictions on trade, while others will permit them when they are reasonable within a specific context. The author proposes that a multifactor test be applied on a case-by-case basis to determine the reasonableness of the no-hire provision in a given employment contract and suggests drafting improvements to facilitate enforcement. PMID- 17402658 TI - "Independence" and the nonprofit board: a general counsel's guide. AB - In the wake of the Sarbanes-Oxley Act regulations that govern the public company sector, standards are emerging to assure that nonprofit corporate boards are maintaining appropriate levels of independence. This Article provides a summation of the current trends in the development of independence standards for nonprofit corporate governance, from both tax and corporate law perspectives. The authors consider independence standards for nonprofit boards of governance and discuss the evolution of independence standards as they relate to the duty of good faith, and the distinction between independence and conflicts of interest. The authors also seek to examine the evolution of current federal regulations and study state models that have been successfully implemented to insure the independence of nonprofit corporations. Finally, the authors propose a set of core guidelines to be considered when addressing board and committee independence issues. PMID- 17402659 TI - Elder abuse detection in nursing facilities: using paid clinical competence to address the nation's shame. AB - Each year, five million senior citizens are the victims of abuse in our country and that figure is a likely underestimate. In California alone, it is estimated that over 225,000 seniors are abused annually. Nursing home residents are particularly vulnerable to abuse, with one-fifth to one-third of these institutions cited for abusive activities that result in actual harm. Regular violations of minimum care regulations by nursing homes have been documented in every state. Despite legal protections, as a practical matter, the review, assessment, and investigation of nursing home care and potential senior abuse is the primary responsibility of voluntary ombudsmen. These volunteers are responsible for regular visits and determination of nursing home quality and resident treatment. This Article contends that, despite their commitment to the cause, voluntary ombudsmen are not a sufficient cure for the scourge of elder abuse. Rather, clinically-trained personnel with legal knowledge must assume the role that lay volunteer ombudsmen are inappropriately expected to play. Use of clinical case managers, who have expertise in assessing clinical factors and can be trained in legal matters, offers significant potential to assist in the detection, and, further, reporting and prosecution of elder abuse. The author concludes that only through use of those with appropriate training and knowledge can the epidemic of elder abuse be effectively addressed. PMID- 17402660 TI - An organization's waiver of the attorney-client privilege and/ or the attorney work product protection to obtain leniency in federal court sentencing: what is the brouhaha all about? AB - The guidelines controlling the sentencing of organizations provide for the reduction in an entity's culpability score for self-reporting, cooperation, and acceptance of responsibility. What an organization must do in order to receive the reduction in culpability score changed dramatically in 2004 when additional language was added to Application Note 12 of the U.S. Sentencing Guidelines Manual Section 8C2.5(g) stating that "waiver of the attorney-client privilege and of work product protections is not a prerequisite to a reduction. ... However, in some circumstances waiver of the attorney-client privilege and of work product protections may be required in order to satisfy the requirements of cooperation." Following months of hearings and public comment, the United States Sentencing Commission reversed its position on whether a sentencing court should consider an organization's waiver of the attorney-client privilege and/or of the attorney work product protection in evaluating the organization's "cooperation" as a sentencing factor by proposing to retract the language added by the 2004 amendments. Although that proposal has become effective, it is yet to be determined what the response of the three branches of government will be on the issue of privilege waivers in the context of federal criminal law. This Article gives readers an overview of the development of the use of privilege waivers by organizations seeking credit for cooperation at the time of sentencing for federal crimes, the reaction of both corporations and their lawyers to the waiver issue, and the events leading up to the Commission's change of position. PMID- 17402661 TI - Whither wundt? PMID- 17402662 TI - Eye centring in portraits: a theoretical and empirical evaluation. AB - Tyler (1998 Nature 392 877) proposed that 'painters centre one eye in portraits', and that this is a hidden aesthetic principle used implicitly by artists and by viewers of portraits. We assess that hypothesis in three related studies: a Monte Carlo analysis of eye placement in synthetic faces randomly placed within a frame; a survey of eye position in 786 painted portraits from Western art of the past six centuries; and an experimental study in which fifty subjects were asked which of two versions of 60 pairs of portraits they preferred, in only one of which the eye was precisely centred. Taken together, the three studies showed no evidence to support Tyler's hypothesis, and in particular there was no evidence for subjects having an aesthetic preference for a centred eye in portraits. We conclude that one eye tends to be relatively close to the vertical midline because of geometric constraints on the placing of a relatively large object, the head, within a pictorial frame. PMID- 17402663 TI - What is in a view? The role of featural information in the recognition of unfamiliar faces across viewpoint transformation. AB - In recognising a face the visual system shows a remarkable ability in overcoming changes in viewpoint. However, the mechanisms involved in solving this complex computational problem, particularly in terms of information processing, have not been clearly defined. Considerable evidence indicates that face recognition involves both featural and configural processing. In this study we examined the contribution of featural information across viewpoint change. Participants were familiarised with unknown faces and were later tested for recognition in complete or part-face format, across changes in view. A striking effect of viewpoint resulting in a reduction in profile recognition compared with the three-quarter and frontal views was found. However, a complete-face over part-face advantage independent of transformation was demonstrated across all views. A hierarchy of feature salience was also demonstrated. Findings are discussed in terms of the problem of object constancy as it applies to faces. PMID- 17402664 TI - The hollow-face illusion: object-specific knowledge, general assumptions or properties of the stimulus? AB - The hollow-face illusion, in which a mask appears as a convex face, is a powerful example of binocular depth inversion occurring with a real object under a wide range of viewing conditions. Explanations of the illusion are reviewed and six experiments reported. In experiment 1 the detrimental effect of figural inversion, evidence for the importance of familiarity, was found for other oriented objects. The inversion effect held for masks lit from the side (experiment 2). The illusion was stronger for a mask rotated by 90 degrees lit from its forehead than from its chin, suggesting that familiar patterns of shading enhance the illusion (experiment 2). There were no effects of light source visibility or any left/right asymmetry (experiment 3). In experiments 4-6 we used a 'virtual' hollow face, with illusion strength quantified by the proportion of noise texture needed to eliminate the illusion. Adding characteristic surface colour enhanced the illusion, consistent with the familiar face pigmentation outweighing additional bottom-up cues (experiment 4). There was no difference between perspective and orthographic projection. Photographic negation reduced, but did not eliminate, the illusion, suggesting shading is important but not essential (experiment 5). Absolute depth was not critical, although a shallower mask was given less extreme convexity ratings (experiment 6). We argue that the illusion arises owing to a convexity preference when the raw data have ambiguous interpretations. However, using a familiar object with typical orientation, shading, and pigmentation greatly enhances the effect. PMID- 17402665 TI - Familiar other-race faces show normal holistic processing and are robust to perceptual stress. AB - Other-race individuals are remembered more poorly and receive less holistic/configural processing than same-race individuals, at least when faces are novel. Here, we examine the amelioration of these effects with familiarity, using distinctiveness-matched Caucasian and Asian stimulus sets. We confirmed a cross-race deficit for upright faces following a single encoding trial, which disappeared rapidly with practice on a small set of other-race 'friends' and did not re-emerge when perceptual processing was put under stress (presentation in the periphery). We also examined holistic/configural processing for familiarised faces using the peripheral inversion effect (McKone, 2004 Journal of Experimental Psychology: Learning, Memory, and Cognition 30 181 - 197). A test for faces and nonface objects (dogs) confirmed the validity of this technique as providing a direct measure of holistic processing; we then showed that, after 1 h of training, holistic processing was as strong for other-race as same-race faces. We conclude that practice with other-race individuals can rapidly engage normal face processing mechanisms. PMID- 17402666 TI - Perceived slant: a dissociation between perception and action. AB - Perceived slant is grossly overestimated, such that 5 degrees hills look to be about 20 degrees. However, overestimation is found only in visual and verbal measures of apparent slant; action measures are accurate. This dissociation is consistent with several lines of research that suggest that there exist two perceptual processes, one for visually guided actions and another for explicit awareness. However, studies in other contexts have shown that analogous effects can be the result of differences in the task demands associated with the responses themselves as opposed to the processes underlying the responses. Two experiments are reported in which these alternatives were tested. Our results are consistent with the hypothesis that two perceptual processes underlie the dissociation between explicit awareness and visuomotor assessments of perceived slant. PMID- 17402667 TI - Perception of visual inclination in a real and simulated urban environment. AB - The perceived inclination of slopes is generally overestimated. We claim that overestimation depends on the use of impoverished stimuli and on the distance between the observer and an inclined surface. In experiment 1, participants reported the perceived inclination of a set of urban roads from two different viewing distances. Observers did not overestimate the perceived inclination of slopes when they saw roads from the shorter viewing distances, whereas they slightly overestimated the perceived inclination of slopes from the farther distance. In experiment 2, participants reported the perceived inclination of a set of stereoscopic slides representing the same urban roads as in experiment 1. Here, observers did not overestimate the perceived inclination of slopes when the projected stereoscopic image contained horizontal disparity and simulated the shorter viewing distance; while they revealed a slight overestimation from the farther distance. We found always overestimation when the binocular image did not contain horizontal disparity, independently from the viewing distance. In conclusion, slopes are overestimated when (a) horizontal disparity is absent, and (b) the viewing distance is increased. PMID- 17402668 TI - Online control of discrete action following visual perturbation. AB - We examined the spatial and temporal limitations of the visual corrective process in goal-directed aiming, as well as gender differences in online control. An initial experiment was conducted to test the utility of a monocular switch procedure as a method of rapidly introducing a visual perturbation. The results revealed minimal effect of the monocular switch on movement time and the endpoint error. Following this control experiment, prismatic displacement was introduced at the initiation of and during the movement. In the third experiment, the prism was presented prior to movement initiation, and then removed at the beginning of or during the movement. Movement trajectories were most influenced by the early presentation and removal of the prism, and female performance was significantly more affected by both perturbations than male performance. PMID- 17402669 TI - The role of voluntary and involuntary attention in selecting perceptual dominance during binocular rivalry. AB - When incompatible images are presented to corresponding regions of each eye, perception alternates between the two monocular views (binocular rivalry). In this study, we have investigated how involuntary (exogenous) and voluntary (endogenous) attention can influence the perceptual dominance of one rival image or the other during contour rivalry. Subjects viewed two orthogonal grating stimuli that were presented to both eyes. Involuntary attention was directed to one of the grating stimuli with a brief change in orientation. After a short period, the cued grating was removed from the image in one eye and the uncued grating was removed from the image in the other eye, generating binocular rivalry. Subjects usually reported dominance of the cued grating during the rivalry period. We found that the influence of the cue declined with the interval between its onset and the onset of binocular rivalry in a manner consistent with the effect of involuntary attention. Finally, we demonstrated that voluntary attention to a grating stimulus could also influence the ongoing changes in perceptual dominance that accompany longer periods of binocular rivalry Voluntary attention did not increase the mean dominance period of the attended grating, but rather decreased the mean dominance period of the non-attended grating. This pattern is analogous to increasing the perceived contrast of the attended grating. These results suggest that the competition during binocular rivalry might be an example of a more general attentional mechanism within the visual system. PMID- 17402670 TI - Prosopagnosia in biographies and autobiographies. AB - Prosopagnosia is a selective impairment of the visual learning and recognition of faces. The congenital type, which is not accompanied by detectable brain damage or malformation, was recently found to be far more common than previously known. Therefore, one should expect that at least a few biographies or autobiographies would reveal a prosopagnosia. In this paper we present an autobiography and a biography describing five cases of congenital prosopagnosia. These biographic descriptions of prosopagnosia add further evidence to the assumption that the congenital type of prosopagnosia is not a rare condition, and not as socially crippling as one might expect. PMID- 17402671 TI - Everything that Fred Astaire did, Ginger Rogers did backwards, and in high heels. PMID- 17402672 TI - A magnetic strategy for new graduate nurses. AB - With the shortfall of nurses, more health care organizations are actively recruiting new graduate nurses. To translate active recruitment to successful retention, however, these newest nurses need support to make the adjustment from the security of a protected academic environment to the demands of a professional nursing career. Nursing turnover occurs when their role transition is unsuccessful. A unique model implemented in 2003 transformed new graduate nurses into confident staff nurses in an acute care setting. The insights gained and the investments required for a "magnetic" strategy to make new graduate nurses successful new staff nurses who stay with the organization are revealed. PMID- 17402673 TI - Return on investment: benefits and challenges of baccalaureate nurse residency program. AB - There are high rates of turnover among graduate nurses within the first year of employment. As the costs of recruitment and orientation have increased, employers are examining new strategies for increasing retention among this population. The Methodist Hospital, Houston, TX, partnered with an academic institution and University Health Consortium to apply an implementation strategy where graduate nurses participate in a year-long residency program. The residents attended monthly sessions that addressed pertinent needs of the group. During this 1-year period turnover decreased from 50% to 13% and return on investment was 326.5%. PMID- 17402674 TI - Foreign-educated nurses: an overview of migration and credentialing issues. AB - It is important for nursing administrators to understand the migration and credentialing process as they integrate foreign-educated nurses into United States health care systems. Issues and considerations related to migration and credentialing of foreign-educated nurses are summarized. PMID- 17402675 TI - An economic analysis of patient simulators clinical training in nursing education. AB - Patient simulators have become widely used in medical education including increasing use in nursing education. Research suggests their usefulness in developing nursing competence. Little research to date, however, has examined the financial feasibility of the use of patient simulators as an educational tool. The extent to which a simulation lab comprising six Laerdal SimMan high-fidelity patient simulators and a staff of nurse educators is a financially feasible alternative to classroom-based education is examined. PMID- 17402676 TI - The impact of nursing on patient and organizational outcomes. AB - The impact of nursing on patient and organizational outcomes has received significant attention from researchers in the past 10 years. Although some research has been based on a theoretical framework, to date, there has not been one, consistent framework that can guide this research. In addition, the frameworks that have been used do not identify nursing as the pivotal concept in the model. The purpose of this article is to identify a framework to guide such research with an emphasis on nursing as the essential concept. Unit culture, nurse staffing, staffing mix, and other variables do not have a direct effect on outcomes. It is only through these variables' influence on the delivery of nursing care that outcomes are affected. PMID- 17402677 TI - Continuing nursing education changes affecting Nursing Economic$. PMID- 17402678 TI - Forging relationships to strengthen recruitment. PMID- 17402679 TI - Social concern: a continuing tradition for nursing. PMID- 17402680 TI - Nursing-sensitive care measures: a platform for value and vision. PMID- 17402681 TI - Staff engagement: it starts with the leader. AB - Engagement can happen if you can take care of yourself first and respect the need for recovery times. It's very hard to find an engaged staff when the leader/manager is disengaged. Engagement is contagious. As leaders model engagement and connection, the staff will follow. Unfortunately, disengagement is also contagious and can create disastrous results. PMID- 17402682 TI - Measuring change: a key component of building a culture of safety. PMID- 17402683 TI - [In vitro cultivation of fields isolates of Plasmodium falciparum in Mali]. AB - Malaria immunology, molecular biology and pathogenicity studies often require the adaptation of Plasmodium falciparum field isolates to continuous in vitro cultivation. For this purpose we have established propagation protocols of asexual erythrocytic stages of P. falciparum samples from malaria patients or asymptomatic carriers in Mali. The parasites were grown in standard culture medium supplemented by human serum and in a culture medium without human serum but supplemented by AlbuMax 1. The candle jar environment and tissue culture flasks gassed with 5% CO2, 5% O2 and 90% N2 obtained from a portable gas mixer were used. Protocols for parasite cultivation in a resource-poor setting were developed. These protocols were successfully applied to fresh isolates in Mali as well as to blood samples frozen in liquid nitrogen and shipped to a laboratory in U.S.A. PMID- 17402684 TI - [Surveillance of falciparum malaria susceptibility to antimalarial drugs and policy change in the Comoros]. AB - Between May and June 2001, efficacy of chloroquine was assessed in 5 sentinel sites in the 3 Comoro islands. Among the 183 children, age range between 6 and 59 months, followed up for 14 days, clinical failure rates ranged between 31.2 and 73.1% and the total failure rate (clinical and parasitological) between 50 and 88.5%. Failures were mainly early treatment failures. The Ministry of health, during a consensus meeting decided to change the first line drug and to gather baseline information on the efficacy and the tolerance of the combination artemether-lumefantrine. Between June and September 2004, among the 164 children, age range between 6 and 59 months included, the success rate of the combination was 99.4% in the 3 sites with a follow-up of 28 days. No serious drug related adverse event was reported. PMID- 17402685 TI - [Cystic echinococcosis in children in Tunisia: fertility and case distribution of hydatid cysts]. AB - Cystic echinococcosis, which commonly starts during childhood or adolescence, is a serious problem of public health in Tunisia. For 121 children (161 cysts), the localization and fertility of cysts as well as viability of their protoscoleces were determined. Results indicated that the lung was the primary localization of cyst (59%) followed by the liver (35%). Children's infection is more frequent in male than in female (sex ratio 1.96) and the greatest number of cases is observed in the 4-9 year age groups (94 cases). The fertility of the cyst was independent of its site or its size and no incidence of age of children was detected. Nevertheless, the fertility rate is higher in females than in males for the liver localization. PMID- 17402686 TI - [Two cases of cryptococcal meningitis revealed by an ischemic stroke]. AB - The usual clinical expression of neuromeningeal cryptococcosis is a meningoencephalitis. We report two cases of neurocryptococcosis which have been revealed by an unusual clinical aspect: an ischemic stroke with a vasculitis mechanism. The two patients had a positive reaction for the HIV and we discussed the responsibility of the HIV or the Cryptococcus in the occurrence of the cerebral infarct. PMID- 17402687 TI - [Simian smallpox (or monkey smallpox): study of 8 cases observed at Impfondo Hospital in Republic of Congo]. AB - Monkeypox or simian smallpox is a viral disease whose clinical manifestations are similar to the human smallpox. In this study authors describe the care management and follow up of eight patients hospitalized at the Impfondo hospital in Congo. It was a transversal study conducted from the 23rd of May till the 7th of July 2003 at the District Hospital of Impfondo (Likouala province, Republic of Congo). We have analyzed 81 patient files hospitalized during that period. Using the Atlanta CDC criteria of case definition modified in 2003, monkeypox has been diagnosed in 8 patients. Our series has 4 male and 4 female patients. The mean age is 9.05 +/- 5.86 with extremes ranging from 5 months to 18 years old. The clinical manifestations included fever (n = 7), rash (n=8), articulations and muscles pains (n=2), mumps (n=4) and pruritus (n=6). 7 patients recovered among them 5 without complications and 2 with persistent hypochromic lesions for more than 6 weeks. The monkeypox strain responsible for these infections appeared not to be lethal to humans. However this study shows that monkeypox is an endemic public health risk in the sanitary district of Impfondo, within the Congo River basin. The endemic presence of monkeypox in the Congo basin could trigger a new outbreak if the viral strains involved here became lethal. PMID- 17402688 TI - Imported melioidosis with an isolated cutaneous presentation in a 90-year-old traveller from Bangladesh. AB - Melioidosis is a tropical disease caused by infection with the bacterium Burkholderia pseudomallei. Most cases present as an acute febrile illness with severe pneumonia and sepsis. Sub-acute and late onset disease can also occur Melioidosis has been diagnosed among travellers who contracted the disease while staying in endemic areas during the rainy season. We report a case of travel associated B. pseudomallei cutaneous infection in a febrile 90-year-old woman with diabetes mellitus, with early stage manifestations of an isolated inoculation lesion. A 32 weeks' treatment with oral amoxicillin-clavulanate and doxycycline combination regimen led to resolution of the lesion and lack of relapse over fifteen months of follow-up. Melioidosis should be considered in the differential diagnosis of unusual subacute cutaneous lesions in a febrile patients returning from endemic areas, as successful management largely depends on early diagnosis and specific long-term suppressive antimicrobial therapy at an early stage of the course of the disease. PMID- 17402689 TI - [Hydrocholecystis, a rare etiology of painful abdominal crisis in sickle cell disease. About two cases]. AB - The authors report two cases of hydocholecystis causing abdominal pains in the sickle cell child. The patients were two girls aged respectively 4 and 12. Hydrocholecystis is defined by acute distension of the gallbladder The diagnosis was made by scan which allowed to follow the evolution in the two sickle cell children. After two crises, no more recurrence was noted in the first patient, on the other hand in the second child recurrences became more frequent. Some authors indicate surgery systematically while others recommend it only after several recurrences. PMID- 17402690 TI - [Portopulmonary hypertension due to schistosomiasis in two Malagasy patients]. AB - Portopulmonary hypertension is characterized by a chronic liver disease associated with a mean pulmonary artery pressure >25 mmHg at rest, an increased pulmonary vascular resistance and a capillary pulmonary pressure <15 mmHg with portal hypertension. Schistosomiasis may be an aetiology of this syndrome, however, few cases have been reported. We describe the first cases of portopulmonary hypertension with schistosomiasis in Malagasy patients. There were 2 men aged of 18 and 20 from hyperendemic area of schistosomiasis in Madagascar Both had a history of repeated water contact. They presented a dyspnea associated with ascites and oedema. Clinical examination showed portal and pulmonary hypertension with right ventricular heart failure. Cardiac examination revealed a systolic murmur and splint of the second heart pulmonary Pulmonary hypertension was confirmed by cardiac ultrasonogaphy Serology of bilharzias was positive. Parasitological examination showed eggs of S. mansoni. The treatment based on salt-free diet, spironolactone and praziquantel led to a better evolution of symptoms (case 1). Symptoms of right heart failure remained for the second patient even though improvement was noted. In tropical countries, schistosomiasis may be one of the cause of portopulmonary hypertension and may appear in early age. Its treatment remains difficult as the drugs recommended are not affordable. PMID- 17402691 TI - [Female genital tuberculosis: about 11 cases treated in Antananarivo (Madagascar)]. AB - A 5 year prospective study on genital tuberculosis, a rather uncommon localization, has been undertaken in 11 women treated at the DAT-HIS of Antananarivo, Madagascar Clinical diagnosis is problematic, if not unfeasible, due to the polymorphism of genital tuberculosis in women. Only histological and bacteriological examinations are, so far, confirmatory In the future, recourse to polymerase chain reaction will facilitate diagnosis and will allow a more accurate assessment of the incidence of this aspect of tuberculosis infection. Genital tuberculosis compromises women's fertility Even though the tuberculosis is cured, none of the 11 women of our research had carried their pregnancy to delivery, because artificial fertilization is not feasible here in Madagascar Screening of woman genital tuberculosis should be mandatory as regards gynaecological problems such as menstrual cycle disorders, sterility, abdominal pain, cyst of ovary ectopic pregnancy, spontaneous miscarriage in paraclinical investigations, especially in developing countries. PMID- 17402692 TI - [The management of stroke in Phnom Penh, Cambodia]. AB - Stroke ranks first among nervous pathologies in Kampuchea. It's a main cause of disability and mortality in our country. We conducted a prospective study including 100 patients hospitalized in the service of general medicine at the Calmette hospital in Phnom Penh. We analyzed the principal risk factors, clinical signs, nature of stroke, complications and markers of the vital and functional prognosis. This work shows the difficulties encountered in the initial care of stroke: delay or absence of hospitalization, cost of complementary examinations to be carried out to determine the nature and the aetiology of stroke and very low level of follow-up to ensure secondary prevention and functional rehabilitation. It can be explained in part by the socioeconomic and cultural level. Research like this one which assesses local needs for stroke prevention, treatment and rehabilitation should be conducted in developing countries to inform the planning and allocation of health care resources in order to reduce the burden of illness associated with stroke. The progressive improvement of the medical structures, and of the socioeconomic and cultural level will facilitate stroke care management. PMID- 17402693 TI - [Clinical polymorphism of cutaneous leishmaniasis in centre and south of Tunisia]. AB - The cutaneous leishmaniasis (CL) is an affection which is quite well known in Tunisia. The zoonotic cutaneous leishmaniasis caused by Leishmania major by far the more frequent, is endemo-epidemic in the centre and south of the country. It is characterized by clinical polymorphism. The aim of our study is to precise the different clinical aspects of the CL in our region through a prospective study of 102 cases. The average age was 37.8 years old (from 4 to 78 years old) with a slight female predominance. All of our cases lived or stayed in an endemic zone. Various clinical forms were noted in our series. The ulcerated and crusted form was predominant: 54,9% of the cases, the lupoid form was noted in 15.7% of the cases and the sporotrichoid form was observed in 18.6% of the cases. Other rare forms were noted (papular erysipeloid, verrucous, vegetant, erythematous, ulcerated, necrotic and linear) were noted in 25.5% of the cases. Our series is characterized by the multiplicity of clinical forms. Besides, the classical form (ulcerated and crusted form), other clinical form can be individualised: lupoid, loco regional spreading (sporotrichoid form, satellite papules). Some atypical forms can be found which are due to variation of host immune responses and to the strain of the parasites involved. PMID- 17402694 TI - [Acanthamoeba keratitis: about the first two Tunisian cases]. AB - Acanthamoeba keratitis is a rare but severe corneal infection which, despite improvements in diagnosis and treatment, still culminates in prolonged morbidity and significant loss of visual acuity. We present the case report of the first identification of Acanthamoeba as a causative agent of keratitis in Tunisia. Case no 1: A 20-year-old girl, nearsighted corrected with soft contact lenses, suffering from a deep corneal inflammation and poor visual acuity The ophthalmological examination showed bilateral dendritiform epithelial keratitis. The illness did not respond to topical and general antibiotic treatment and developed bilateral corneal abscess. Microscopic examination and culture of samples from cornea scraping revealed the presence of trophozoit and cysts of Acanthamoeba associated with Fusarium oxysporum. As the treatment with local Ketoconazol and antibiotherapy didn't show any result, two transfixiant keratoplasty were carried out and treatment by Desomedine, PHMB (polyhexamethylene biguanide) and Voriconazol was started. After two months, the patient felt better, vision was also improved (2/10) and infiltrates became smaller Case no 2: A 19-year-old girl, nearsighted with soft contact lenses consulted for a bilateral corneal ulceration and poor vision (1/20). Trophozoit and cysts of Acanthamoeba were found in the contact lens solution. She was treated quickly with Desomedine. Visual acuity improved to 7/10 but the corneal ulceration left a residual opacity PMID- 17402695 TI - [Predictive factors of dengue shock syndrome at the children Hospital No. 1, Ho chi-Minh City, Vietnam]. AB - The dengue shock syndrome (DSS) is primarily a complication of dengue haemorrhagic fever (DHF) among children in South East Asia. A case-control study was carried out at the children hospital no 1 (Ho-Chi-Minh City, Vietnam) in May July 2005, to identify the predictive factors of the DSS among 1-15 year patients with DHE Forty consecutive admitted cases and forty controls were studied. The associated features of DSS were the 7-12 year age group and the re-infection by the dengue virus. The vaccination against the Japanese encephalitis B was not associated statistically significantly with the shock syndrome. The clinical predictors of DSS gathered an abdominal tenderness, an hepatomegaly, a lethargy, a cold extremity presentation. DSS associated laboratory features were a value of hematocrit a 50 % and a platelet cell count < or = 75,000/mm3. PMID- 17402696 TI - [Canine visceral leishmaniasis in northeast Brazil: epidemiological aspects]. AB - In a rural area of Northeast Brazil, the relatively high serological infection by Leishmania in dogs, the lack of classical vector Lutzomyia longipalpis and of American Visceral Leishmaniasis cases in human beings and the observation of Leishmania in ticks collected in infected dogs suggest that ticks may be responsible for the transmission between dogs. PMID- 17402697 TI - [Tuberculosis and HIV/AIDS co-infection in children: experience carried out in paediatric service of the teaching hospital of Brazzaville, Republic of Congo (1995-2004)]. AB - Epidemiologic, clinical and paraclinical data of 126 children with tuberculosis whose HIV status was known, have been compared. Among them, 65% were HIV positive, the co-infection tuberculosis HIV/AIDS was observed in all social categories. The source of contamination was discovered for 72% of the patients. The mother was involved in 47.5% of cases. The main reasons of consultation were a long standing fever a chronic cough and a weight loss. Diarrhea was mainly observed in positive HIV patients (p = 0.00). The general condition was influenced by a weight loss which was more important in positive patients with a IMC lower than 10 in 12.8% of cases. There was no significant difference between all clinical forms. Digital hippocratism, chronic otitis and parotiditis were only observed in positive HIV patients with skin illness ten times more frequent (p = 0.00). Anergia to tuberculin tests (78.4%) and a sedimentation speed up to 100 mm at the first hour were observed in more than 60% of the positive HIV patients (p = 0.001). Evolution was favorable under treatment for 84% of positive HIV patients with an increasing weight becoming weaker after one month of treatment. All deaths happened among that population. PMID- 17402698 TI - [Salmonella meningitis in Ouagadougou, Burkina Faso, from 2000 to 2004]. AB - 56 strains of Salmonella were isolated from the cerebro-spinal fluids (CSF) from meningitis suspected patients at the Yalgado-Ouedraogo University hospital center in Burkina Faso, from January 2000 to December 2004. 75% of the patients were less than 3 years old; 71.4% of the CSF were purulent, with an average of 523 leucocytes/mm3 and 78% of neutrophile polynuclears. The strains identified belonged mostly to Salmonella O: 4.5 group (51.8%). In vitro, 92.7% of the strains were resistant to ampicillin and this resistance was partially restored with amoxicillin/clavulanic acid; however no strain was resistant to ceftriaxone. For the overall 56 patients, 20 different antibiotherapy regimes were used and they were successful in only 27% cases while 71% of patients died and 2% escaped from the hospital. Neurologic sequels were found in a patient treated with both ceftriaxone and chloramphenicol. These results showed that the illness occurred mainly in infants and was associated with high mortality rate. Most of the Salmonella strains were multi-drug resistant. In spite of strains multi antibiotics resistance, adequate definition of therapeutic lines and early treatment including ceftriaxone could lead to higher cure rates and may improve the outcome. PMID- 17402699 TI - [Seroprevalence of Helicobacter pylori infection in Malagasy population]. AB - Helicobacter pylori is a worldwide infection. However very few data are actually available on H. pylori seroprevalence in the Malagasy population. We carried out a transversal study in a sample of persons who met the following criteria: older than 15 years old, presence in the medicine internal unit 2 (University Hospital Center of Antananarivo) during the period of the study whatever the reason. H. pylori infection was identified serologically by using ELISA (G.A.P IgG H. pylori ELISA, Bio-Rad, France). Several factors were evaluated including serological status, demographic information, the reason of the presence in the unit, factors influencing H. pylori infection: socio-economic status, siblings, promiscuity consumption of alcohol, use of tobacco, water source and history of gastroscopy. The presence of clinical symptoms, such as dyspepsia and abdominal pain, was determined. Forty-five men and 45 women were included (mean age: 41.8 +/- 3.4 years). The seroprevalence of H. pylori infection was 82%. H. pylori infection was higher in men than in women (p < 0.02). Promiscuity constituted the principal factor influencing H. pylori infection. The seroprevalence of the H. pylori infection appears to be comparable to the rate encountered in developing countries. Considering this high rate of the H. pylori infection, eradication of H. pylori should be commonly recommended when facing gastrointestinal pathologies potentially induced by H. pylori. PMID- 17402700 TI - [Human auricular myiasis caused by Wohlfahrtia magnifica (Diptera: Sarcophagidae): about three observations in Morocco]. AB - We report three cases of human otomyiasis observed in rural Moroccan children. Myiasis of external orifices usually occurs from neglected chronic lesions of the patients with poor personal hygiene. The parasitologic identification revealed Wohlfahrtia magnifica. Wohlfahrtiosis is common myiasis of sheep and goats in Mediterranean basin. Through this paper we underline the epidemiological, pathogenic, clinical and therapeutic aspects of this parasitosis. PMID- 17402701 TI - Alkyne creation in aliphatic polymers: influence of side groups. AB - In this article, we focus on the influence of side groups on terminal alkyne creation, in aliphatic polymers submitted to swift heavy ions, under vacuum. Terminal alkyne creation was compared in polyethylene and in substituted polymers. We selected two classes of side groups: alkyl groups, which differed in their length (polypropylene and polybutene) and allyl groups, which were linear (EPDMh) or cyclic (EPDMn). Irradiated samples were analyzed using Fourier transform IR spectroscopy, in the transmission mode, at room temperature. Alkynes are created when the electronic stopping power, (dE/dx)e, exceeds a threshold value. This threshold value was moderately influenced by the presence of an alkyl side chain but was clearly reduced in the presence of C=C bonds on the side chain. Nevertheless, in all-saturated polymers, below the (dE/dx)e threshold, terminal alkyne formation is observed after a latent dose, rather independently of the side-chain length and directly related to the formation of radiation induced double bonds prior to alkyne formation. Under S ion irradiation, the radiation chemical yield G0 value obtained in EPDMh was 4 times the value of G0 in PE. This effect is understandable only if important energy transfers, from the backbone to C=C double bonds, are considered. PMID- 17402702 TI - Synthesis and self-assembly of amphiphilic polymeric microparticles. AB - We report the synthesis and self-assembly of amphiphilic, nonspherical, polymeric microparticles. Wedge-shaped particles bearing segregated hydrophilic and hydrophobic sections were synthesized in a microfludic channel by polymerizing across laminar coflowing streams of hydrophilic and hydrophobic polymers using continuous flow lithography (CFL). Particle monodispersity was characterized by measuring both the size of the particles formed and the extent of amphiphilicity. The coefficient of variation (COV) was found to be less than 2.5% in all measured dimensions. Particle structure was further characterized by measuring the curvature of the interface between the sections and the extent of cross-linking using FTIR spectroscopy. The amphiphilic particles were allowed to self-assemble in water or at water-oil interfaces. In water, the geometry of the particles enabled the formation of micelle-like structures, while in emulsions, the particles migrated to the oil-water interface and oriented themselves to minimize their surface energy. PMID- 17402703 TI - Proton-actuated membrane-destabilizing polyion complex micelles. AB - The efficiency of nucleic acid-based drugs is usually hampered by the fact that, following their uptake by the cell, these drugs end up in acidic organelles (i.e., endosomes/lysosomes) from which they barely escape. This work relates to the preparation and characterization of polyion complex micelles (PICM) formed by the self-assembly of three polyelectrolytes: a diblock cationic copolymer; a membranolytic, methacrylic acid copolymer; and an oligonucleotide. It is demonstrated that a synthetic membrane-active polyanion can be successfully integrated within the structure of PICM to yield well-defined, narrowly distributed micelles (30 nm) with a core/shell architecture. Besides their ability to protect the oligonucleotide against nuclease degradation, PICM partly dissociate under mildly acidic conditions, releasing chain clusters that destabilize bilayer membranes. This association/dissociation behavior illustrates the potential of these pH-sensitive PICM for the transport and efficient delivery of polyionic drugs. PMID- 17402704 TI - Synthesis of enzyme-degradable, peptide-cross-linked dextran hydrogels. AB - Hydrogels derived from synthetic polymers have been previously engineered to degrade under the activity of matrix metalloproteinases (MMPs). It is believed that these systems can act as extracellular-matrix (ECM) equivalents mimicking the degradation and remodeling of the ECM through the activity of cell-secreted enzymes. In this study, MMP-sensitive hydrogels derived from dextran were developed. In order to avoid the incorporation of hydrolyzable esters often introduced in dextran modification strategies, the polysaccharide was modified with p-maleimidophenyl isocyanate (PMPI) thereby introducing maleimide functionalities in the backbone and resulting in dextran derivatized with p maleimidophenyl isocyanate (Dex-PMPI). This strategy was favored to separate out the effects of random hydrolysis and enzymatic digestion in the degradation of the dextran hydrogels. A peptide cross-linker, derived from collagen and susceptible to gelatinase A (MMP-2) digestion, was synthesized with bifunctional cysteine termini and used to cross-link the Dex-PMPI. These hydrogels were found to be hydrolytically stable for more than 200 days yet degraded either within 30 h when exposed to bacterial collagenase or within 16 days when exposed to human MMP-2, demonstrating enzymatic-mediated digestion of the peptide cross-links. Further modification of the cross-linked hydrogels with laminin-derived peptides enhanced cell adhesion and survival, demonstrating the potential of these materials for use in tissue engineering applications. PMID- 17402705 TI - The same drug but a different mechanism of action: comparison of free doxorubicin with two different N-(2-hydroxypropyl)methacrylamide copolymer-bound doxorubicin conjugates in EL-4 cancer cell line. AB - Doxorubicin is one of the most potent anti-tumor drugs with a broad spectrum of use. To reduce its toxic effect and improve its pharmacokinetics, we conjugated it to an HPMA copolymer carrier that enhances its passive accumulation within solid tumors via the EPR effect and decreases its cytotoxicity to normal, noncancer cells. In this study, we compared the antiproliferative, pro-survival, and death signals triggered in EL-4 cancer cells exposed to free doxorubicin and doxorubicin conjugated to a HPMA copolymer carrier via either enzymatically (PK1) or hydrolytically (HYD) degradable bonds. We have previously shown that the intracellular distribution of free doxorubicin, HYD, and PK1 is markedly different. Here, we demonstrated that these three agents greatly differ also in the antiproliferative effect and cell death signals they trigger. JNK phosphorylation sharply increased in cells treated with HYD, while treatment with free doxorubicin moderately decreased and treatment with PK1 even strongly decreased it. On the other hand, treatment with free doxorubicin greatly increased p38 phosphorylation, while PK1 and HYD increased it slightly. PK1 also significantly increased ERK phosphorylation, while both the free doxorubicin and HYD conjugate slightly decreased it. Long-term inhibition of JNK significantly increased both proliferation and viability of EL-4 cells treated with free doxorubicin, showing that the JNK signaling pathway could be critical for mediating cell death in EL-4 cells exposed to free doxorubicin. Both activation of caspase 3 and decreased binding activity of the p50 subunit of NFkappaB were observed in cells treated with free doxorubicin and HYD, while no such effects were seen in cells incubated with PK1. Analysis of the expression of genes involved in apoptosis and regulation of the cell cycle demonstrated that free doxorubicin and HYD have very similar mechanisms of action, while PK1 has very different characteristics. PMID- 17402706 TI - Quaternized poly(4-vinylpyridine)s as model gene delivery polycations: structure function study by modification of side chain hydrophobicity and degree of alkylation. AB - To optimize polycation-based gene delivery agents, the influence of molecular characteristics of the polycations on physicochemical properties of polycation/DNA complexes and their relationships to cellular gene transfer need to be understood. With this aim, we have synthesized a series of model polymers based on quaternized poly(4-vinylpyridine)s (CnPVP-beta) with the same DP of 1600 but differing by the number n of methylene groups in N-alkyl ester substituents from 1 to 6 and/or by degree of alkylation beta from 25% to 95%. For polycations CnPVP-95, the efficiency of transfection of a plasmid vector expressing a secreted form of alkaline phosphatase started to be detectable at n = 4, noticeable at n = 5, and again undetectable at n = 6. A decrease in beta of active C5PVP-95 from 95% to 65% resulted in a further noticeable increase of activity with a 9-fold increase for C5PVP-65. This finding was attributed to the proton sponge mechanism due to protonation of non-alkylated pyridine moieties of CnPVP-beta/DNA complexes in slightly acidic media that was supported by the fluorescence quenching assay. The data demonstrate the advantages of partial alkylation of tertiary polyamines with medium-length alkyl agents for preparation of efficient nonviral gene delivery vectors. PMID- 17402707 TI - Conjugation of a new two-photon fluorophore to poly(ethylenimine) for gene delivery imaging. AB - We report herein the molecular engineering of an efficient two-photon absorbing (TPA) chromophore based on a donor-donor bis-stilbenyl entity to allow conjugation with biologically relevant molecules. The dye has been functionalized using an isothiocyanate moiety to conjugate it with the amine functions of poly(ethylenimine) (PEI), which is a cationic polymer commonly used for nonviral gene delivery. Upon conjugation, the basic architecture and photophysical properties of the active TPA chromophore remain unchanged. At the usual N/P ratio (ratio of the PEI positive charges to the DNA negative charges) of 10 used for transfection, the transfection efficiency and cytotoxicity of the labeled PEI/DNA complexes were found to be comparable to those of the unlabeled PEI/DNA complexes. Moreover, when used in combination with unlabeled PEI (at a ratio of 1 labeled PEI to 3 unlabeled PEI), the labeled PEI does not affect the size of the complexes with DNA. The labeled PEI was successfully used in two-photon fluorescence correlation spectroscopy measurements, showing that at N/P = 10 most PEI molecules are free and the diffusion coefficient of the complexes is consistent with the 360 nm size measured by quasielastic light scattering. Finally, two-photon images of the labeled PEI/DNA complexes confirmed that the complexes enter into the cytoplasm of HeLa cells by endocytosis and hardly escape from the endosomes. As a consequence, the functionalized TPA chromophore appears to be an adequate tool to label the numerous polyamines used in nonviral gene delivery and characterize their complexes with DNA in two-photon applications. PMID- 17402708 TI - Liquid chromatography-tandem mass spectrometry application, for the determination of extracellular hepatotoxins in Irish lake and drinking waters. AB - A novel method for the determination of hepatotoxins; microcystins (MCs), and nodularin (Nod) in lake water and domestic chlorinated tap water has been developed using liquid chromatography hyphenated with electrospray ionization triple quadrupole mass spectrometry (LC-ESI-MS/MS). Optimization of the mass spectrometer parameters and mobile-phase composition was performed to maximize the sensitivity and reproducibility of the method. Detection of the hepatotoxins was carried out using multiple reaction monitoring experiments, thus improving the selectivity of the method. A total ion chromatogram and a precursor ion scan on ion m/z 135 was also applied to all samples to detect unknown microcystins or microcystins for which there are no standards available. A comprehensive validation of the LC-ESI-MS/MS method was completed that took into account matrix effects, specificity, linearity, accuracy, and precision. Good linear calibrations were obtained for MC-LR (1-200 microg/L; R2=0.9994) in spiked lake and tap water samples (1-50 microg/L; R2=0.9974). Acceptable interday repeatability was achieved for MC-LR in lake water with RSD values (n=9) ranging from 9.9 (10 microg/L) to 5.1% (100 microg/L). Excellent limits of detection (LOD) and limits of quantitation (LOQ) were achieved with spiked MCs and Nod samples; LOD=0.27 microg/L and LOQ=0.90 microg/L for MC-LR in the "normal linear range" and LOD=0.08 microg/L and LOQ=0.25 microg/L in the "low linear range" in both lake and chlorinated tap water. Similar results were obtained for a suite of microcystins and nodularin. This sensitive and rapid method does not require any sample preconcentration, including the elimination of solid-phase extraction (SPE) for the effective screening of hepatotoxins in water below the 1 microg/L WHO provisional guideline limit for MC-LR. Furthermore, SPE techniques are time consuming, nonreproducible at trace levels, and offer poor recoveries with chlorinated water. The application of this LC-ESI-MS/MS method for routine screening of hepatotoxins in lake and chlorinated tap water (average Cl2=0.23 mg/L) is achieved and this study represents the first direct method for the screening of hepatotoxins in chlorinated tap water. PMID- 17402709 TI - Functionalized magnetic nanoparticles for small-molecule isolation, identification, and quantification. AB - Functionalized magnetic nanoparticles (MNPs) were synthesized to serve as laser desorption/ionization elements as well as solid-phase extraction probes for simultaneous enrichment and detection of small molecules in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis. Two laser-absorbing matrices were each conjugated onto MNP to give MNP@matrix which provided high ionization efficiency and background-free detection in MS leading to unambiguous identification of target small molecules in a complex mixture. MNP@matrix was demonstrated to serve as a general matrix-free additive in MALDI-TOF MS analysis of structurally distinct small molecules. Also, MNP@matrix provides a simple, rapid, and reliable quantitative assay for small molecules by mass without either the use of an internal standard or an isotopic labeling tag. Furthermore, the affinity extraction of small molecules from complex biofluid was achieved by probe protein-conjugated MNP@matrix without laborious purification. We demonstrated that a nanoprobe-based assay is a cost effective, rapid, and accurate platform for robotic screening of small molecules. PMID- 17402710 TI - Understanding the changes in the circular dichroism of light harvesting complex II upon varying its pigment composition and organization. AB - In this work we modeled the circular dichroism (CD) spectrum of LHCII, the main light harvesting antenna of photosystem II of higher plants. Excitonic calculations are performed for a monomeric subunit, taken from the crystal structure of trimeric LHCII from spinach [Liu, Z. F., Yan, H. C., Wang, K. B., Kuang, T. Y., Zhang, J. P., Gui, L. L., An, X. M., and Chang, W. R. (2004) Nature 428, 287-292]. All of the major features of the CD spectrum above 450 nm are satisfactorily reproduced, and possible orientations of the Chl and carotenoid transition dipole moments are identified. The obtained modeling parameters are used to simulate the CD spectra of two complexes with altered pigment composition: a mutant lacking Chls a 611-612 and a complex lacking the carotenoid neoxanthin. By removing the relevant pigment(s) from the structure, we are able to reproduce their spectra, which implies that the alteration does not disturb the overall structure. The CD spectrum of trimeric LHCII shows a reversed relative intensity of the two negative bands around 470 and 490 nm as compared to monomeric LHCII. The simulations reproduce this reversal, indicating that it is mainly due to interactions between chromophores in different monomeric subunits, and the trimerization does not induce observable changes in the monomeric structure. Our simulated spectrum resembles one of two different trimeric CD spectra reported in literature. We argue that the differences in the experimental trimeric CD spectra are caused by changes in the strength of the monomer-monomer interactions due to the differences in detergents used for the purification of the complexes. PMID- 17402711 TI - Crystal structures of acetylcholinesterase in complex with organophosphorus compounds suggest that the acyl pocket modulates the aging reaction by precluding the formation of the trigonal bipyramidal transition state. AB - Organophosphorus compounds (OPs), such as nerve agents and a group of insecticides, irreversibly inhibit the enzyme acetylcholinesterase (AChE) by a rapid phosphorylation of the catalytic Ser203 residue. The formed AChE-OP conjugate subsequently undergoes an elimination reaction, termed aging, that results in an enzyme completely resistant to oxime-mediated reactivation by medical antidotes. In this study, we present crystal structures of the non-aged and aged complexes between Mus musculus AChE (mAChE) and the nerve agents sarin, VX, and diisopropyl fluorophosphate (DFP) and the OP-based insecticides methamidophos (MeP) and fenamiphos (FeP). Non-aged conjugates of MeP, sarin, and FeP and aged conjugates of MeP, sarin, and VX are very similar to the noninhibited apo conformation of AChE. A minor structural change in the side chain of His447 is observed in the non-aged conjugate of VX. In contrast, an extensive rearrangement of the acyl loop region (residues 287-299) is observed in the non-aged structure of DFP and in the aged structures of DFP and FeP. In the case of FeP, the relatively large substituents of the phosphorus atom are reorganized during aging, providing a structural support of an aging reaction that proceeds through a nucleophilic attack on the phosphorus atom. The FeP aging rate constant is 14 times lower than the corresponding constant for the structurally related OP insecticide MeP, suggesting that tight steric constraints of the acyl pocket loop preclude the formation of a trigonal bipyramidal intermediate. PMID- 17402712 TI - Rational design of substituted N-alkoxypyridine-2(1H)thiones with increased stability against daylight. AB - N-alkoxypyridine-2(1H)thiones serve as valuable photochemical alkoxyl radical precursors in photobiological studies, but due to a broad absorption band at about 360 nm (pi --> pi* excitation), these molecules decompose readily when exposed to daylight. The goal of the present work is to propose N-alkoxypyridine 2(1H)thiones which due to a blue shift of this band become more stable with respect to daylight and consequently are easier to handle. The shift of the pi - > pi* excitation toward shorter wave length shall be achieved by substituents introduced at the pyridine heterocycle. To study the substituent effects, excitations to the first to singlet states were calculated applying the CASPT2 approach and time dependent density functional theory (TD-DFT). The study indeed showed that electron rich substituents (like the methoxylgroup) at the positions 3, 4, and 6 of the pyridinethione heterocycle yield the desired hypsochromic shift. A free rotation of the substituent, however, is expected to quench these effects. Fluorine atoms, employed to model the influence of electron withdrawing substituents, induce also a blue shift for a substitution at the 3, 4, and 6 positions. For the multiply fluorinated molecule N-methoxy-3,4,6 trifluorinepyridine-2(1H)thione a blue shift of even 24 nm is predicted. Substituents that can conjugate with the pi electrons of the heterocycle (NO2 served as a model) only induce strong bathochromic shifts on the pi --> pi* excitation energy and therefore are not able to eliminate the daylight sensitivity of the precursor molecules. PMID- 17402713 TI - Oligothiophene catenanes and knots. Part II. Mono and dications. A theoretical study. AB - Mono- and dications of catenanes and knots containing 16, 22, and 28 thiophene units have been studied at the BHandHLYP/3-21G* level of theory. The polaron localization and relaxation energies of monoionized molecules increase with dihedral angle between thiophene fragments, being higher for catenanes and knots compared to linear oligomers. Monoionization of catenanes results in the polaron localization at one macrocycle leaving another one intact. In all diionized oligomers, polaron pairs were found to be more stable than corresponding bipolarons. The energy difference between bipolaron and polaron pairs increases with the number of repeating units in oligomers for all studied molecular architectures. Singlet polaron pairs are more stable than triplet ones. The energy difference between triplet and singlet states does not exceed 7-8 kcal/mol and decreases with the number of thiophene units in oligomers. Two different singlet minima were found for diionized catenanes. In the first one (the most stable), each macrocycle loses one electron, and in the other one, the polaron pairs are located at one macrocycle, leaving another intact. The energy difference between two minima decreases with the number of repeating units in catenanes. PMID- 17402714 TI - Laplacian field of the effectively unpaired electron density: determination of many-body effects on electron distributions. AB - This work carries out the study of the Laplacian field function of the electron density L(r) = -nabla2rho(r) splitted in two contributions rho(r) = rho(p)(r) + rho(u)(r), which correspond to the effectively paired and effectively unpaired electron densities, respectively. The visualization of the concentration and depletion of these fields and their spatial localization show no contribution of the effectively unpaired electrons to the conventional bonding among two centers, but the field -nabla2rho(u)(r) provides an interesting structure. We also study the reliability of the information contained in the partitioning of this electron density field function for describing nonclassical bondings as the three-center two-electron ones. PMID- 17402715 TI - Dielectric properties of ethyleneglycol-1,4-dioxane mixtures using TDR method. AB - Complex permittivity has been determined for mixtures of ethyleneglycol-1,4 dioxane (EG-DX) with various concentrations in the frequency range from 100 MHz to 30 GHz at 25 degrees C by time domain reflectometry (TDR). A primary process with an asymmetric shape and a Debye-type small-amplitude high-frequency process are observed for each mixture. The deviation of the relaxation time for the primary process from that of the ideal mixture shows a maximum value at a mole fraction of 1,4-dioxane, xDX approximately =0.8. The static permittivity for the mixtures can be explained using the Luzar model by assuming the formation of two types of hydrogen-bonded dimers, one between EG-EG (pair 1) and the other between EG-DX (pair 2). The number of these pairs is also estimated as a function of concentration. These results of the relaxation time and static permittivity are interpreted on the basis of a model of two kinds of cooperative domains coexisting in the mixtures. PMID- 17402716 TI - Molecular dynamics simulations of the solution-air interface of aqueous sodium nitrate. AB - Molecular dynamics simulations have been used to investigate the behavior of aqueous sodium nitrate in interfacial environments. Polarizable potentials for the water molecules and the nitrate ion in solution were employed. Calculated surface tension data at several concentrations are in good agreement with measured surface tension data. The surface potential of NaNO3 solutions at two concentrations also compare favorably with experimental measurements. Density profiles suggest that NO3- resides primarily below the surface of the solutions over a wide range of concentrations. When the nitrate anions approach the surface of the solution, they are significantly undercoordinated compared to in the bulk, and this may be important for reactions where solvent cage effects play a role such as photochemical processes. Surface water orientation is perturbed by the presence of nitrate ions, and this has implications for experimental studies that probe interfacial water orientation. Nitrate ions near the surface also have a preferred orientation that places the oxygen atoms in the plane of the interface. PMID- 17402717 TI - Thermochemistry of key soot formation intermediates: C3H3 isomers. AB - Accurate standard enthalpies of formation for allene, propyne, and four C3H3 isomers involved in soot formation mechanisms have been determined through systematic focal point extrapolations of ab initio energies. Auxiliary corrections have been applied for anharmonic zero-point vibrational energy, core electron correlation, the diagonal Born-Oppenheimer correction (DBOC), and scalar relativistic effects. Electron correlation has been accounted for via second order Z-averaged perturbation theory (ZAPT2) and primarily through coupled cluster theory, including single, double, and triple excitations, as well as a perturbative treatment of connected quadruple excitations [ROCCSD, ROCCSD(T), ROCCSDT, and UCCSDT(Q)]. The correlation-consistent hierarchy of basis sets, cc pVXZ (X = D, T, Q, 5, 6), was employed. The CCSDT(Q) corrections do not exceed 0.12 kcal mol(-)1 for the relative energies of the systems considered here, indicating a high degree of electron correlation convergence in the present results. Our recommended values for the enthalpies of formation are as follows: Delta(f)H(o)(0)(propargyl) = 84.76, Delta(f)H(o)(0) (1-propynyl) = 126.60, Delta(f)H(o)(0) (cycloprop-1-enyl) = 126.28, Delta(f)H(o)(0)(cycloprop-2-enyl) = 117.36, Delta(f)H(o)(0)(allene) = 47.41, and Delta(f)H(o)(0)(propyne) = 46.33 kcal mol(-1), with estimated errors no larger than 0.3 kcal mol(-1). The corresponding C3H3 isomerization energies are about 1 kcal mol(-1) larger than previous coupled-cluster results and several kcal mol(-1) below those previously obtained using density functional theory. PMID- 17402718 TI - Structure and dynamics of beta-cyclodextrin in aqueous solution at the density functional tight binding level. AB - Structure and dynamics of beta-cyclodextrin (beta-CyD), a prototype host for inclusion compounds of biological interest, is investigated by means of density functional based tight-binding molecular dynamics (MD) simulations. The computational protocol is benchmarked against available experimental data and first-principles calculations. Solvent-solute interactions, including the diffusion into and dwell time of the solvent in the cavity of beta-CyD, are studied with a hybrid QM/MM method. Comparison of MD simulations of beta-CyD in the gas phase and in water shows that the solvent reduces the flexibility of the structure framework, while the terminal hydroxyl groups become more flexible and are embedded in a network of hydrogen bonds. Our 160 ps MD simulations, provide enough sampling to discuss the dynamics of the water inside the cavity. The dwell time of the encapsulated water molecule has a wide distribution with a peak at 70 fs. Surprisingly, despite only the 17% difference between the "top" and "bottom" opening area of the beta-CyD cone, 64% of the water molecules enter the cavity through the slightly bigger "bottom" aperture. PMID- 17402719 TI - Proton affinity correlations between hydrogen and dihydrogen bond acceptors. AB - Several series of hydrogen- and dihydrogen-bonded complexes with HCN, C2H2, HF, H2O, CH3CONH2, and CH3COOH as donors and H2O, MeOH, EtOH, MeOMe, NH3, NH2Me, NHMe2, NMe3, NEtMe2, and BH3-NMe3 as acceptors were investigated using the MP2/6 311++G(d,p) level of theory. The total lowering of the X-H stretching frequencies in the hydrogen-bonded complexes were linearly correlated with the proton affinities of the accepting bases. From comparison of hydrogen- and dihydrogen bonded complexes, a scaling factor to estimate the exact proton affinity of a dihydrogen bond acceptor was developed. Further, the scaling factor involving linear donors (1.204) is marginally higher than that involving nonlinear donor molecules (1.162). Finally, it was found that, given identical conditions, a hydrogen bond will be about 16-20% stronger than a corresponding dihydrogen bond. PMID- 17402720 TI - Structure-activity relationships for NAMI-A-type complexes (HL)[trans-RuCl4L(S dmso)ruthenate(III)] (L = imidazole, indazole, 1,2,4-triazole, 4-amino-1,2,4 triazole, and 1-methyl-1,2,4-triazole): aquation, redox properties, protein binding, and antiproliferative activity. AB - Imidazolium [trans-tetrachloro(1H-imidazole)(S-dimethylsulfoxide)ruthenate(III)] (NAMI-A) and indazolium [trans-tetrachlorobis(1H-indazole)ruthenate(III)] (KP1019) are the most promising ruthenium complexes for anticancer chemotherapy. In this study, the azole ligand of NAMI-A was systematically varied (from imidazole of NAMI-A to indazole, 1,2,4-triazole, 4-amino-1,2,4-triazole, and 1 methyl-1,2,4-triazole), and the respective complexes were evaluated with regard to the rate of aquation and protein binding, redox potentials, and cytotoxicity by means of capillary zone electrophoresis, electrospray ionization mass spectrometry, cyclic voltammetry, and colorimetric microculture assays. Stability studies demonstrated low stability of the complexes at pH 7.4 and 37 degrees C and a high reactivity toward proteins (binding rate constants in the ranges of 0.02-0.34 and 0.01-0.26 min-1 for albumin and transferrin, respectively). The redox potentials (between 0.25 and 0.35 V) were found to be biologically accessible for activation of the complexes in the tumor, and the indazole containing compound shows the highest antiproliferative activity in vitro. PMID- 17402721 TI - Dihydropyridopyrazinones and dihydropteridinones as corticotropin-releasing factor-1 receptor antagonists: structure-activity relationships and computational modeling. AB - The CRF antagonist pharmacophore is a heterocyclic ring bearing a critical hydrogen-bond acceptor nitrogen and an orthogonal aromatic ring. CRFR1 antagonists have shown a 40-fold and 200-fold loss in potency against the CRFR1 H199V and M276I mutant receptors, suggesting key interactions with these residues. We have derived a two component computational model that correlates CRFR1 binding affinity within the reported series to antagoinst/H199 complexation energy and M276 hydrophobic contacts. PMID- 17402722 TI - Investigation of the lactam side chain length necessary for optimal indenoisoquinoline topoisomerase I inhibition and cytotoxicity in human cancer cell cultures. AB - Indenoisoquinolines with lactam substituents such as ethylamino, propylamino, and butylamino have previously demonstrated potent biological activity, but an optimal length has never been established. In the present study, a series of simplified indenoisoquinoline analogues possessing a linker spacing of 0-12 carbon atoms between the lactam nitrogen and the terminal amino group have been prepared, determining that 2-4-atom lengths are optimal for topoisomerase I inhibition and cytotoxicity. Using these lengths, analogues were prepared with the amino group and portions of the linker replaced by a pyridine ring. A three carbon spacer within the pyridine series still demonstrated potent topoisomerase I inhibition. PMID- 17402723 TI - Structure-activity relationship studies of gonadotropin-releasing hormone antagonists containing S-aryl/alkyl norcysteines and their oxidized derivatives. AB - A series of acyline analogues incorporating l- and d-isomers of S arylated/alkylated norcysteines [Ncy(R), where R is 2-naphthyl, methyl, and isopropyl] at positions 1, 4, 7, and 10 were synthesized. Some of these analogues were mono- and dioxidized to sulfoxides and sulfones. All of the analogues of acyline were screened for the antagonism of the GnRH-induced response in a reporter gene assay in HEK-293 cells expressing the human GnRH receptor. Nine of the analogues (9, 11, 15, 16, 17, 19, 20, 21, and 22) had antagonistic potency (IC50 < 2 nM) similar to that of acyline (IC50 = 0.52 nM) in this assay. Selected analogues (9, 11, 15, 16, 19, and 21) were tested in vitro for their antagonism at the rat GnRH-R in a reporter gene assay as well as in an in vivo intact male rat assay. Analogues 9 and 15 were the most potent in suppressing testosterone levels. PMID- 17402724 TI - Identification of anthranilic acid derivatives as a novel class of allosteric inhibitors of hepatitis C NS5B polymerase. AB - A series of potent anthranilic acid-based inhibitors of the hepatitis C NS5B polymerase has been identified. The inhibitors bind to a site on NS5B between the thumb and palm regions adjacent to the active site as determined by X-ray crystallography of the enzyme-inhibitor complex. Guided by both molecular modeling and traditional SAR, the enzyme activity of the initial hit was improved by approximately 100-fold, yielding a series of potent and selective NS5B inhibitors with IC50 values as low as 10 nM. These compounds were also inhibitors of the HCV replicon in cultured HUH7 cells. PMID- 17402725 TI - Indium-labeled macrocyclic conjugates of naltrindole: high-affinity radioligands for in vivo studies of peripheral delta opioid receptors. AB - We have identified a series of hydrophilic indium-labeled DOTA and DO3A conjugates of naltrindole (NTI) that are suited to in vivo studies of peripheral delta opioid receptors. Indium(III) complexes, linked to the indole nitrogen of NTI by six- to nine-atom spacers, display high affinities (0.1-0.2 nM) and excellent selectivities for binding to delta sites in vitro. The [111In]-labeled complexes can be prepared in good isolated yields ( approximately 65%) with high specific radioactivities (>3300 mCi/mumol). The spacers serve as pharmacokinetic modifiers, and log D7.4 values range from -2.74 to -1.79. These radioligands exhibit a high level of specific binding (75-94%) to delta opioid receptors in mouse gut, heart, spleen, and pancreas in vivo. Uptakes of radioactivity are saturable by the non-radioactive complexes, inhibited by naltrexone, and blocked by NTI. Thus, these radiometal-labeled NTI analogues warrant further study by single-photon emission computed tomography. PMID- 17402726 TI - 9-(2-C-Cyano-2-deoxy-beta-D-arabino-pentofuranosyl)guanine, a potential antitumor agent against B-lymphoma infected with Kaposi's sarcoma-associated herpesvirus. AB - Several 9-(2-C-cyano-2-deoxy-l-beta-d-arabino-pentofuranosyl)purine derivatives were tested against Kaposi's sarcoma-associated herpesvirus (KSHV)-infected primary effusion lymphoma (PEL) cells. The guanine derivative (3, CNDAG), as well as the 2-amino-6-substituted-purine derivatives 4, 5, and 6, exhibited cell growth inhibitory activity against KSHV-infected cells, but showed no cytotoxicity against KSHV-negative cells at >15 microM concentrations. Therefore, it was found that compounds 3, 4, 5, and 6 showed selective cytotoxicity against PEL cells infected with KSHV. PMID- 17402727 TI - New methacrylate-functionalized ba and ba-ti oxoclusters as potential nanosized building blocks for inorganic-organic hybrid materials: synthesis and characterization. AB - Two different methacrylate modified barium-titanium and barium-based oxoclusters, Ba2Ti10(mu3-O)8(mu2-OH)5(mu2-OMc)20(OiPrOMe)2 (1) and [Ba(OMc)2(McOH)3]n (2), were synthesized by reacting methacrylic acid with barium-titanium and barium zirconium double alkoxides, respectively. The X-ray structure determination of oxocluster 1 shows a core consisting of a ring of 10 titania octahedra, sharing corners, that surround the two barium oxygen decaeders which are linked by common edges to the titania octahedra and the neighboring barium decaeder. The solid state structure of 2 consists of zigzag chains of edge-sharing {BaO9} polyhedra linked through bridging bidentate metacrylate anions, displaying different coordination mode of carboxylate groups. The presence of methacrylate groups surrounding the two polynuclear compounds has been exploited for the embedding of the oxocluster in inorganic-organic hybrid materials, and some preliminary results are presented. PMID- 17402728 TI - The first ruthenium-based paullones: syntheses, X-ray diffraction structures, and spectroscopic and antiproliferative properties in vitro. AB - Two novel paullone derivatives, namely, 6-(alpha-picolylamino)-7,12 dihydroindolo[3,2-d][1]benzazepine (L1) and 9-bromo-6-(alpha-picolylamino)-7,12 dihydroindolo[3,2-d][1]benzazepine (L2), have been prepared. The reaction of cis [RuCl2(DMSO)4] (DMSO=dimethyl sulfoxide) with L1 and L2 in a 1:1 molar ratio in dry ethanol at 50 degrees C afforded the complexes trans-[RuIICl2(DMSO)2L1] (1a) and trans-[RuIICl2(DMSO)2L2] (1b) in 26 and 30% yield, respectively. The reaction carried out from the same starting compounds in a 1:2 molar ratio at 75 degrees C led to the formation of [RuIICl(DMSO)(L1)2]Cl (2a) and [RuIICl(DMSO)(L2)2]Cl (2b) in 16 and 23% yield, correspondingly. The products were characterized by elemental analysis, one- and two-dimensional NMR spectroscopy, electrospray ionization mass spectrometry, IR spectroscopy, electronic spectra, cyclic voltammetry, and X-ray crystallography (L1, L2, 1a, and 2b). Complexes 2a and 2b exhibit remarkable antiproliferative activity in three human carcinoma cell lines, A549 (non-small cell lung carcinoma), CH1 (ovarian carcinoma), and SW480 (colon carcinoma). The novel complexes show an intercalative mode of interaction with DNA, which may render them attractive alternatives to metal compounds with a coordinative mode of interaction. PMID- 17402729 TI - Synthesis, reactivity, and structural characterization of sodium and ytterbium complexes containing new imidazolidine-bridged bis(phenolato) ligands. AB - A new imidazolidine-bridged bis(phenol) [ONNO]H2 ([ONNO]H2=1,4-bis(2-hydroxy-3,5 di-tert-butyl-benzyl)imidazolidine) was prepared in relatively high yield by Mannish reaction of 2,4-di-tert-butylphenol, formaldehyde, and ethylenediamine in a 2:3:1 molar ratio. Reaction of the bis(phenol) with NaH in THF, after workup, afforded the sodium bis(phenolate) {[ONNO]Na2(THF)2}2.2THF (1) as a dimeric tetranuclear complex in an almost quantitative yield. Reaction of YbCl3 with complex 1 in a 2:1 molar ratio in THF, in the presence of HMPA, produced the desired bis(phenolate) ytterbium dichloride as bimetallic complex [ONNO]{YbCl2(HMPA)}2.2.5C7H8 (2). Complex 2 can be used as a precursor for the synthesis of ytterbium derivatives by salt metathesis reactions. Reaction of complex 2 with NaOiPr in a 1:2 molar ratio in THF led to the formation of bimetallic alkoxide [ONNO]{Yb(mu-OiPr)Cl(HMPA)}2.THF (3). However, the residual chlorine atoms in complex 3 are inactive for the further substituted reaction. Further study revealed that the bulkiness of the reagent has profound effect on the outcome of the reaction. Complex 2 reacted with bulky NaOAr (ArO=2,6-di-tert butyl-4-methylphenoxo) or NaNPh2 in a 1:2 molar ratio under the same reaction conditions, after workup, to give the ligand redistributed products, (ArO)2YbCl(HMPA)2 (4) and [ONNO]YbCl(HMPA)2 (5) for the former and complexes 5 and (Ph2N)2YbCl(HMPA)2 (6) for the latter. If the molar ratio of complex 2 to NaNPh2 decreased to 1:4, the expected ligand redistributed products [ONNO]YbNPh2(HMPA) (7) and (Ph2N)3Yb(HMPA)2.C7H8 (8) can be isolated in high yields. All of the complexes were well characterized, and the definitive molecular structures of complexes 1-4, 7, and 8 were provided by single-crystal X ray analysis. PMID- 17402730 TI - Photosensitive function of encapsulated dye in carbon nanotubes. AB - Single-wall carbon nanotubes (SWCNTs) exhibit resonant absorption localized in specific spectral regions. To expand the light spectrum that can be utilized by SWCNTs, we have encapsulated squarylium dye into SWCNTs and clarified its microscopic structure and photosensitizing function. X-ray diffraction and polarization-resolved optical absorption measurements revealed that the encapsulated dye molecules are located at an off center position inside the tubes and aligned to the nanotube axis. Efficient energy transfer from the encapsulated dye to SWCNTs was clearly observed in the photoluminescence spectra. Enhancement of transient absorption saturation in the S1 state of the semiconducting SWCNTs was detected after the photoexcitation of the encapsulated dye, which indicates that ultrafast (<190 fs) energy transfer occurred from the dye to the SWCNTs. PMID- 17402731 TI - Networks of coupled rotators: relationship between structures and internal dynamics in metal-binding proteins. Applications to apo- and holo-calbindin. AB - This article presents an analysis of the internal dynamics of the Ca2+-binding protein calbindin, based on the Networks of Coupled Rotators (NCRs) introduced recently. Several fundamental and practical issues raised by this approach are investigated. The roles of various parameters of the model are examined. The NCR model is shown to account for the modifications of the internal dynamics upon Ca2+ binding by calbindin. Two alternative strategies to estimate local internal effective correlation times of the protein are proposed, which offer good agreement between predictions and experiment. PMID- 17402732 TI - Bifunctional binding of cisplatin to DNA: why does cisplatin form 1,2-intrastrand cross-links with ag but not with GA? AB - The bifunctional binding of the anticancer drug cisplatin to two adjacent nucleobases in DNA is modeled using density functional theory. Previous experimental studies revealed that cisplatin binding to adjacent guanine and adenine is sensitive to nucleobase sequence. Whereas AG 1,2-intrastrand cross links are commonly observed, the analogous GA adducts are not known. This study focuses on understanding this directional preference by constructing a full reaction profile using quantum chemical simulation methods. Monofunctional and bifunctional cisplatin adducts were generated, and the transition states that connect them were located for the dinucleotides d(pApG) and d(pGpA), assuming that initial platination takes place at the guanine site. Our computer simulations reveal a significant kinetic preference for formation of the AG over the GA adduct. The activation free energies of approximately 23 kcal/mol for AG and approximately 32 kcal/mol for GA suggest that bifunctional closure is approximately 6 orders of magnitude faster for AG than for GA. A strong hydrogen bond between one of the ammine ligands of cisplatin and the 5' phosphate group of the DNA backbone is responsible for the stabilization of the transition state that affords the AG adduct. This interaction is absent in the transition state that leads to the GA adduct because the right-handed helix of the DNA backbone places the phosphate out of reach for the ammine ligand. We found only an insignificant thermodynamic difference between AG and GA adducts and conclude that the preference of AG over GA binding is largely under kinetic control. The puckering of the deoxyribose ring plays an important role in determining the energetics of the bifunctional platination products. Whereas the 3'-nucleoside remains in the native C2'-endo/C3'-exo form of B-DNA, the deoxyribose of the 5' nucleoside always adopts the C2'-exo/C3'-endo puckering in our simulations. A detailed analysis of the energies and structures of the bifunctional adducts revealed that the observed sugar puckering patterns are necessary for platinum to bind in a relaxed coordination geometry. PMID- 17402733 TI - Kinetic and computational analysis of the palladium(II)-catalyzed asymmetric allylic trichloroacetimidate rearrangement: development of a model for enantioselectivity. AB - The asymmetric rearrangement of allylic trichloroacetimidates catalyzed by palladium(II) complexes of the COP family is a powerful method for the preparation of enantioenriched chiral allylic amines from prochiral allylic alcohols. A detailed kinetic analysis of this reaction was performed to elucidate the rate- and enantiodetermining step of this important reaction. The results of these studies support a cyclization-induced rearrangement mechanism and prompted DFT studies (B3LYP/LACVP**+) of C-N bond formation, believed to be the enantiodetermining step of this catalytic cycle. On the basis of these calculations, a model for enantioinduction was developed, in which the planar chirality of the catalyst controls the enantioselectivity. These studies should allow the rational design of more enantioselective catalysts. PMID- 17402734 TI - Distinguishing among structural ensembles of the GB1 peptide: REMD simulations and NMR experiments. PMID- 17402735 TI - Large ground-state entropy changes for hydrogen atom transfer reactions of iron complexes. AB - Reported herein are the hydrogen atom transfer (HAT) reactions of two closely related dicationic iron tris(alpha-diimine) complexes. FeII(H2bip) (iron(II) tris[2,2'-bi-1,4,5,6-tetrahydropyrimidine]diperchlorate) and FeII(H2bim) (iron(II) tris[2,2'-bi-2-imidazoline]diperchlorate) both transfer H* to TEMPO (2,2,6,6-tetramethyl-1-piperidinoxyl) to yield the hydroxylamine, TEMPO-H, and the respective deprotonated iron(III) species, FeIII(Hbip) or FeIII(Hbim). The ground-state thermodynamic parameters in MeCN were determined for both systems using both static and kinetic measurements. For FeII(H2bip) + TEMPO, DeltaG degrees = -0.3 +/- 0.2 kcal mol-1, DeltaH degrees = -9.4 +/- 0.6 kcal mol-1, and DeltaS degrees = -30 +/- 2 cal mol-1 K-1. For FeII(H2bim) + TEMPO, DeltaG degrees = 5.0 +/- 0.2 kcal mol-1, DeltaH degrees = -4.1 +/- 0.9 kcal mol-1, and DeltaS degrees = -30 +/- 3 cal mol-1 K-1. The large entropy changes for these reactions, |TDeltaS degrees | = 9 kcal mol-1 at 298 K, are exceptions to the traditional assumption that DeltaS degrees approximately 0 for simple HAT reactions. Various studies indicate that hydrogen bonding, solvent effects, ion pairing, and iron spin equilibria do not make major contributions to the observed DeltaS degrees HAT. Instead, this effect arises primarily from changes in vibrational entropy upon oxidation of the iron center. Measurement of the electron-transfer half reaction entropy, |DeltaS degrees Fe(H2bim)/ET| = 29 +/- 3 cal mol-1 K-1, is consistent with a vibrational origin. This conclusion is supported by UHF/6-31G* calculations on the simplified reaction [FeII(H2N=CHCH=NH2)2(H2bim)]2+...ONH2 left arrow over right arrow [FeII(H2N=CHCH=NH2)2(Hbim)]2+...HONH2. The discovery that DeltaS degrees HAT can deviate significantly from zero has important implications on the study of HAT and proton-coupled electron-transfer (PCET) reactions. For instance, these results indicate that free energies, rather than enthalpies, should be used to estimate the driving force for HAT when transition metal centers are involved. PMID- 17402737 TI - Coordination-driven nanosized lanthanide "molecular lantern" with tunable luminescent properties. PMID- 17402738 TI - A synthetic nucleoside probe that discerns a DNA adduct from unmodified DNA. PMID- 17402739 TI - Morphology control of fluorescent nanoaggregates by co-self-assembly of wedge- and dumbbell-shaped amphiphilic perylene bisimides. PMID- 17402740 TI - Cation-dependent compression behavior in low-silica zeolite-X. PMID- 17402741 TI - Asymmetric conjugate alkenylation of enones catalyzed by chiral diols. PMID- 17402742 TI - Synthesis and isomerization of imino-fused N-confused porphyrin. AB - [reaction: see text] A set of mutually interconvertable inner-bridged-type porphyrinoids, imino-fused N-confused porphyrins (IF-NCPs), which possess a [5.7.5] tricyclic ring in the core, were synthesized from a condensation reaction of 21-amino-substituted NCP and an arylaldehyde, and the structures were characterized by X-ray single-crystal analysis. PMID- 17402743 TI - Lewis acid catalyzed reaction of arylvinylidenecyclopropanes with ethyl (arylimino)acetates: a facile synthetic protocol for pyrrolidine and 1,2,3,4 tetrahydroquinoline derivatives. AB - [reaction: see text] A number of pyrrolidine and 1,2,3,4-tetrahydroquinoline derivatives are prepared selectively in moderate to good yields by the reaction of arylvinylidenecyclopropanes 1 with ethyl (arylimino)acetates 2 in the presence of Lewis acid depending on the electronic nature both of 2 and R1 or R2 aromatic groups of 1. PMID- 17402744 TI - Changes in color and odorant compounds during oxidative aging of Pedro Ximenez sweet wines. AB - Pedro Ximenez sweet wines obtained following the typical criaderas and solera method for sherry wines and subjected to oxidative aging for 0, 1.3, 4.2, 7.0, or 11.5 years were studied in terms of color and aroma fraction by using the CIELab method and gas chromatography, respectively. The parameters defining the CIELab color space (a*, b*, and L*) were subjected to a multiple-range test (p<0.05) that allowed discrimination of the five wine aging levels studied into five uniform groups according to aging time. The aroma fraction was found to include 15 active odorant compounds with OAV > 1 that enriched the wines with fruity, fatty, floral, and balsamic notes during the aging process. The changes in color parameters and active odorants were not linearly related to aging time, being especially marked during the first 1.3 years and then less substantial up to the 7 years, the oldest wines exhibiting sensorial properties markedly departing from all others. For the wines aged over 1.3 years (minimum aging), 2,3-butanedione, linalool, and decanal can be used as reliable fingerprints of the older wines' quality. PMID- 17402745 TI - Detection methods for biotech cotton MON 15985 and MON 88913 by PCR. AB - Plants derived through agricultural biotechnology, or genetically modified organisms (GMOs), may affect human health and ecological environment. A living GMO is also called a living modified organism (LMO). Biotech cotton is a GMO in food or feed and also an LMO in the environment. Recently, two varieties of biotech cotton, MON 15985 and MON 88913, were developed by Monsanto Co. The detection method is an essential element for the GMO labeling system or LMO management of biotech plants. In this paper, two primer pairs and probes were designed for specific amplification of 116 and 120 bp PCR products from MON 15985 and MON 88913, respectively, with no amplification from any other biotech cotton. Limits of detection of the qualitative method were all 0.05% for MON 15985 and MON 88913. The quantitative method was developed using a TaqMan real-time PCR. A synthetic plasmid, as a reference molecule, was constructed from a taxon-specific DNA sequence of cotton and two construct-specific DNA sequences of MON 15985 and MON 88913. The quantitative method was validated using six samples that contained levels of biotech cotton mixed with conventional cotton ranging from 0.1 to 10.0%. As a result, the biases from the true value and the relative deviations were all within the range of +/-20%. Limits of quantitation of the quantitative method were all 0.1%. Consequently, it is reported that the proposed detection methods were applicable for qualitative and quantitative analyses for biotech cotton MON 15985 and MON 88913. PMID- 17402746 TI - The role of Ets transcription factors in the basal transcription of the translocator protein (18 kDa). AB - The translocator protein (18 kDa; TSPO), previously known as peripheral-type benzodiazepine receptor, is a high-affinity cholesterol- and drug-binding mitochondrial protein involved in various cell functions including steroidogenesis, apoptosis, and proliferation. TSPO is highly expressed in secretory and glandular tissues, especially in steroidogenic cells, and its expression is altered in certain pathological conditions such as cancer and neurological diseases. In this study, we characterized the regulatory elements present in the region of the TPSO promoter extending from 515 to 805 bp upstream of the transcription start site, an area previously identified as being important for transcription. Promoter fragments extending 2.7 kb and 805 bp upstream of the transcription start site were able to direct enhanced green fluorescent protein expression to Leydig cells of the testis, theca cells of the ovary, and cells of the adrenal cortex in transgenic animals. This expression pattern perfectly mimicked endogenous TSPO expression. Functional characterization of the 515-805 bp region revealed the presence of one specificity protein 1/specificity protein 3 (Sp1/Sp3) and two v-ets erythroblastosis virus E26 oncogene homologue (Ets) binding sites that are important for transcriptional activity in both MA-10 mouse Leydig tumor cells and NIH/3T3 whole mouse embryo fibroblasts. GA-binding protein alpha (GABPalpha), a member of the Ets family of transcription factors, was found to be associated with the endogenous TSPO promoter. We conclude that Sp1/Sp3 and members of the Ets family of transcription factors bind to specific binding sites in the TSPO promoter to drive basal TSPO gene transcription. PMID- 17402747 TI - Initial-rate kinetics of human NMN-adenylyltransferases: substrate and metal ion specificity, inhibition by products and multisubstrate analogues, and isozyme contributions to NAD+ biosynthesis. AB - Initial-rate and product inhibition studies revealed distinctive ordered ternary complex kinetic mechanisms, substrate specificities, and metal ion preferences for the three isozymes of human nicotinamide mononucleotide adenylyl-transferase (NMNAT, EC 2.7.7.1). ATP binds before NMN with nuclear isozyme NMNAT1 and Golgi apparatus NMNAT2, but the opposite order is observed with the mitochondrial isozyme NMNAT3. Only the latter utilizes ITP efficiently in place of ATP, and while NMNH conversion to NADH by NMNAT1 and NMNAT3 occurs at similar rates, conversion by NMNAT2 is much slower. These isozymes can also be discriminated by their action on tiazofurin monophosphate (TrMP), a metabolite of the antineoplastic prodrug tiazofurin. Our finding that TrMP is only a substrate with NMNAT1 and NMNAT3 reveals for the first time an organelle selectivity in the metabolism of this important drug. In search of additional ways to discriminate these isozymes, we synthesized and tested the P1-(nicotinamide/nicotinate riboside-5')-Pn-(adenosine-5') dinucleotides Np3AD, Np4AD, and Nap4AD. In addition to being highly effective inhibitors, these multisubstrate geometric inhibitors gave inhibition patterns that are consistent with the aforementioned isozyme differences in substrate binding order. Distinctive differences in their substrate specificity and metal ion selectivity also permitted us to quantify individual isozyme contributions to NAD+ formation in human cell extracts. PMID- 17402748 TI - Positive aspects of negative design: simultaneous selection of specificity and interaction stability. AB - The energetic determinants that drive specific protein-protein interactions are not entirely understood. We describe simultaneous in vivo selection of specific and stable interactions using homologous peptides which compete with protein libraries for an interaction with a target molecule. Library members binding to their target, and promoting cell growth, must outcompete competitor interactions with the target (i.e., competition) and evade binding to the competitors (i.e., negative design). We term this a competitive and negative design initiative (CANDI). We combined CANDI with a protein-fragment complementation assay (PCA) and observed major specificity improvements, by driving selection of winning library members that bind their target with maximum efficacy, ensuring that otherwise energetically accessible alternatives are inaccessible. CANDI-PCA has been used with libraries targeted at coiled coil regions of oncogenic AP-1 components cJun and cFos. We demonstrate that comparable hydrophobic and electrostatic contributions in desired species are compromised in nondesired species when CANDI is executed, demonstrating that both core and electrostatic residues are required to direct specific interactions. Major energetic differences (>or=5.6 kcal/mol) are observed between desired and nondesired interaction stabilities for a CANDI-PCA derived peptide relative to a conventional PCA derived helix, with significantly more stability (3.2 kcal/mol) than the wild-type cJun-cFos complex. As a negative control, a library lacking a residue repertoire able to generate a specific and stable helix was tested. Negative protein design has broad implications in generating specific and therapeutically relevant peptide-based drugs, proteins able to act with minimal cross-talk to homologues or analogues, and in nanobiotechnological design. PMID- 17402749 TI - Screening and identification of GSH-trapped reactive metabolites using hybrid triple quadruple linear ion trap mass spectrometry. AB - The present study describes a new analytical approach for the detection and characterization of GSH-trapped reactive metabolites using multiple reaction monitoring (MRM) as the survey scan to trigger the acquisition of enhanced product ion (EPI) spectra on a triple quadrupole linear ion mass spectrometer. The MRM scan step was carried out following up to 114 MRM transitions from the protonated molecules of potential GSH adducts to their product ions derived from a neutral loss of 129 or 307 Da. MRM transition protocols were constructed on the basis of common bioactivation reactions predicted to occur in human liver microsomes (HLM). The effectiveness and reliability of the approach were evaluated using acetaminophen, diclofenac, and carbamazepine as model compounds. The total ion chromatograms of the MRM for the HLM incubations with these compounds and GSH clearly displayed a number of GSH adducts, including acetaminophen-GSH adducts and carbamazepine-GSH adducts that were not previously observed in HLM incubations. In addition, clomipramine and mefenamic acid that have the frame structures susceptible to P450-mediated bioactivation were investigated. As a result, the MRM-EPI analysis revealed multiple GSH adducts of clomipramine and mefenamic acid in HLM incubations possibly mediated by epoxide and/or quinone imine intermediates. Compared with the neutral loss (NL) and precursor ion (PI) scanning analysis, the MRM-based approach provided superior sensitivity and selectivity for GSH adducts. It also enabled the sensitive acquisition of EPI spectra with rich fragmentation in the same LC/MS run, which were useful for the rapid structure elucidation of GSH adducts and the elimination of false positives. The MRM-EPI experiment can be employed for high throughput screening of reactive metabolites and should be especially applicable to compounds of the same chemotype. Also, it can be applied in conjunction with the PI or NL scan as a comprehensive method for the analysis of reactive metabolites in a drug discovery setting. PMID- 17402750 TI - Future of toxicology--iron chelators and differing modes of action and toxicity: the changing face of iron chelation therapy. AB - Iron (Fe) chelation therapy was initially designed to alleviate the toxic effects of excess Fe evident in Fe-overload diseases. However, the novel toxicological properties of some Fe chelator-metal complexes have shifted appreciable focus to their application in cancer chemotherapy. Redox-inactive Fe chelator complexes are well suited for the treatment of Fe-overload diseases, whereas Fe chelator complexes with high redox activity have shown promising results as chemotherapeutics against cancer. Within this perspective, we discuss the different modes of action and toxicological profiles of Fe chelators, including analogues of 2-pyridylcarboxaldehyde isonicotinoyl hydrazone, di-2-pyridylketone isonicotinoyl hydrazone, di-2-pyridylketone thiosemicarbazone, and the clinically trialed chelator 3-aminopyridine-2-carboxaldehyde thiosemicarbazone. The potential application of these agents in the changing face of Fe chelation therapy is discussed. PMID- 17402751 TI - Epitaxial growth of single-crystalline Al(2)O(3) films on Cr(2)O(3)(0001). AB - Thin, crystallographically oriented single-crystalline Al2O3 films can be grown epitaxially on Cr2O3(0001) by codeposition of Al vapor and O2 at a substrate temperature of 825 K. The properties and growth of these films were monitored by Auger electron spectroscopy (AES), low-energy electron diffraction (LEED), low energy ion scattering (LEIS), and X-ray photoelectron spectroscopy (XPS). Two routes of preparation were investigated: (i) stepwise growth by alternating deposition of Al at room temperature and subsequent exposure to O2 at elevated temperatures; (ii) codeposition of Al and O2 at T > 800 K. The first route was consistently found to result in the growth of a complex interfacial oxide followed by the growth of polycrystalline Al2O3. The second mode of preparation provided homogeneous and ordered, probably (0001)-oriented, films of Al2O3 that maintained a LEED pattern up to a thickness around 10 A. The surface sensitive Cr MVV Auger transition at 34 eV was completely attenuated once the Al2O3 layer had reached a thickness of 6 A, pointing to film homogeneity at an early stage. This was confirmed by the absence of a significant Cr signal in LEIS spectra. PMID- 17402752 TI - Biotinylation of TiO(2) nanoparticles and their conjugation with streptavidin. AB - Photoactive TiO(2) can be used to mediate a variety of disinfection processes. It was postulated that TiO(2) particles could be directed to specific targets of interest using biotin/streptavidin linkages. Biotinylated TiO(2) nanoparticles (anatase) were obtained by treating TiO(2) nanoparticles with 3 aminopropyltriethoxysilane (APTS) in anhydrous DMSO, followed by reaction with N hydroxysuccinimidobiotin. 29Si CP-MAS NMR, 13C CP-MAS NMR, and FTIR spectra showed that biotin was covalently bound to the TiO(2) surface. Transmission electron microscopy (TEM) demonstrated that prolonging the silanization reaction times led to increasingly thick silsesquioxane coating layers of up to approximately 10 nm. The specific surface area (SSA) of the TiO2 particles decreased from 16 m(2) g(-1) before treatment to 9.1 m(2) g(-1) after aminosilanization and to 8.4 m(2) g(-)1 after biotinylation, as measured by nitrogen adsorption. Amino surfaces modified for 4, 16, and 26 h had total amino group densities ranging from 2.9 to 26 to 66 nm(-2), respectively, whereas accessible surface amino group densities ranged from 2.7 to 10 to 17 nm(-2) as shown from nitrogen adsorption, polyelectrolyte titration, conductometric titration, and biotin assays. Not all the amino groups were accessible for biotinylation: the densities of active biotin were found to be 2.1, 7.0, and 11.5 nm(-2). The ability of the attached biotin to bind to streptavidin was demonstrated by confocal microscopy with the use of fluorescently labeled streptavidin-FITC. Although streptavidin was readily able to bind to biotinylated TiO(2) particles, it did not act as a strong flocculating agent for the biotinylated TiO2 particles. The implications of these observations, with respect to particle accessibility to tethered streptavidin, are discussed. PMID- 17402753 TI - Determination of kinetic parameters for interfacial enzymatic reactions on self assembled monolayers. AB - This paper reports a method to characterize the kinetic constants for the action of enzymes on immobilized substrates. This example uses cutinase, a serine esterase that hydrolyzes 4-hydroxyphenyl valerate moieties that are immobilized on a self-assembled monolayer of alkanethiolates on gold. The product of the enzyme reaction is a hydroquinone, which is redox active and therefore permits the use of cyclic voltammetry to monitor the extent of reaction in situ. A kinetic model based on the Michaelis-Menten formalism is used to analyze the dependence of initial rates of reaction on both the substrate density and the enzyme concentration. The resulting value of k(cat)/K(M) for the interfacial reaction is comparable to that for a homogeneous phase reaction with a substrate of similar structure. This strategy of using monolayers presenting substrates for the enzyme and cyclic voltammetry to measure reaction rates provides quantitative and real-time information on reaction rates and permits a level of analysis of interfacial enzyme reactions that to date has been difficult to realize. PMID- 17402754 TI - Two-dimensional nanoparticle arrays derived from ferritin monolayers. AB - A scalable technique for making silica coatings with embedded two-dimensional arrays of iron oxide nanoparticles is presented. The iron oxide nanoparticle arrays were formed by depositing quasi-crystalline ferritin layers, an iron storage protein with an iron oxide mineral core, on solid substrates by a spread coating technique based on evaporation-induced convective assembly. The layer of protein molecular arrays was then encapsulated in a silica matrix film deposited from a sol precursor. The organic protein shell of the ferritin molecules was then removed by controlled pyrolysis, leaving ordered iron oxide cores bound in the silica matrix. This article is the first report on combining convective self assembly of proteins with sol-gel techniques of oxide film formation. The technique is technologically feasible and scalable to make coatings of encapsulated ordered magnetic clusters tens of cm(2) or larger in size. PMID- 17402755 TI - Protein crystallization at the air/water interface induced by shearing bulk flow. AB - We have observed 2D protein crystallization under conditions where in the absence of flow, crystallization fails to occur. Even under conditions where crystallization does occur in quiescent systems, we have found that flow can accelerate the crystallization process. By interrogating the flow responsible for this enhanced crystallization, we have correlated the enhancement with large shear in the plane of the interface. Some possible mechanisms for why interfacial shear can enhance the crystallization process are proposed. PMID- 17402756 TI - Atomistic simulations to compute surface properties of poly(N-vinyl-2 pyrrolidone) (PVP) and blends of PVP/chitosan. AB - Atomistic simulations were performed on poly(N-vinyl-2-pyrrolidone) (PVP) and its blends with chitosan (CS) in different ratios using molecular mechanics (MM) and molecular dynamics (MD) simulations in three-dimensionally periodic and effective two-dimensionally periodic condensed phases. Four independent microstructures were generated to analyze their surface properties. The calculated surface-energy values for PVP compared quite well with the experimental data reported in the literature. The density profile was analyzed, and the structure of the films showed an interior region of the bulk density. Various components of the energetic interactions (torsional, van der Waals, etc.) were examined to gain deeper insight into the nature of regular and anomalous interactions between the bulk and the surface films. Surface energies of PVP/CS blends were computed by MD simulations using the bulk pressure-volume-temperature (PVT) parameters. Bulk properties such as the cohesive energy density (CED) and solubility parameter (delta) were calculated using MM and MD simulations in the NVT ensemble under periodic boundary conditions. The Flory equation of state was used to compute the thermal expansion coefficient as well as PVT parameters. These surface-energy values agreed well with the surface-energy data calculated using the Zisman equation, which were also in accordance with the experimental observations. The results from this study suggest that computer simulations would provide valuable information on polymers and polymer-blend surfaces. PMID- 17402757 TI - Nanostructured Pt/GC model electrodes prepared by the deposition of metal-salt loaded micelles. AB - Novel, nanostructured, carbon-supported Pt model electrodes with homogeneously distributed Pt nanoparticles of uniform size were fabricated and analyzed with respect to their electrochemical properties. For this purpose, Pt-salt-loaded micelles were deposited on a glassy carbon substrate and subsequently exposed to an oxygen plasma and a H2 atmosphere for removal of the polymer carriers and reduction of the Pt salt. The morphology of the resulting nanoparticles and their electrochemical/electrocatalytic properties were characterized by high-resolution scanning electron microscopy, X-ray photoelectron spectroscopy, cyclic voltammetry, and differential electrochemical mass spectrometry for CO electrooxidation. The data demonstrate that this method is generally suited to the production of nanostructured model electrodes with well-defined and independently adjustable particle size and interparticle distance distributions, which are specifically suited for quantitative studies of transport processes in electrocatalytic reactions. PMID- 17402758 TI - Individual electrophoretic mobilities of liposomes and acidic organelles displaying pH gradients across their membranes. AB - This report focuses on measuring the individual electrophoretic mobilities of liposomes with different pH gradients across their membrane using capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). The results from the individual analysis of liposomes show that, using surface electrostatic theories and the electrokinetic theory as the first approximation, zeta potential contributes more significantly to the electrophoretic mobility of liposomes than liposomal size. For liposomes with an outer pH 7.4 (pH(o) 7.4) and a net negative outer surface charge, the most negative electrophoretic mobilities occur when the inner pH (pH(i)) is 6.8; at higher or lower pH(i), the electrophoretic mobilities are less negative. The theories mentioned above cannot explain these pH-induced electrophoretic mobility shifts. The capacity theory, predicting an induced electrical charge on the surface of liposomes, can only explain the results at pH(i) > 6.8. In this report, we hypothesize that there is a flip-flop process of phospholipids, which refers to the exchange of phospholipids between the outer and inner layers of the membrane. This flip-flop is caused by the pH gradient and membrane instability and results in the observed electrophoretic mobility changes when pH(i) is <6.8. Furthermore, it is found that the mobilities of acidic organelles are consistent with the predictions of liposome models we used here. PMID- 17402759 TI - Protein refolding assisted by periodic mesoporous organosilicas. AB - Herein we report a new strategy for protein refolding by taking advantage of the unique surface and pore characteristics of ethylene-bridged periodic mesoporous organosilica (PMO), which can effectively entrap unfolded proteins and assist refolding by controlled release into the refolding buffer. Hen egg white lysozyme was used as a model protein to demonstrate the new method of protein refolding. Through loading of denatured proteins inside uniform mesoporous channels tailored to accommodate individual protein, protein aggregation was minimized, and the folding rate was increased. Poly(ethyleneglycol) (PEG)-triggered continuous release of entrapped denatured lysozyme allowed high-yield refolding with high cumulative protein concentrations. The new method enhances the oxidative refolding of lysozyme (e.g., over 80% refolding yield at about 0.6 mg/mL). PMID- 17402760 TI - Fast dynamics of wormlike micellar solutions. AB - We present the first measurements of the fast dynamics of cationic wormlike micelles (WLM) using neutron spin echo (NSE). The comparison with theory [Zilman, A.; Granek, R. Phys. Rev. Lett. 1996, 77, 4788. Granek, R. J. Phys. II 1997, 7, 1761]1,2 enables coarse grained parameters to be identified. We propose and validate a calibration procedure to extract the bending constant kappa from NSE measurements. PMID- 17402761 TI - Formation and aqueous surface wettability of polysiloxane nanofibers prepared via surface initiated, vapor-phase polymerization of organotrichlorosilanes. AB - The formation of high aspect ratio organosiloxane fibers of nanodimensionality via the surface-induced vapor-phase polymerization of vinyltrichlorosilane is reported. We also demonstrate the versatility of our method by producing fibers of various densities, polydispersities, and lengths. Nanofibers were characterized using SEM, EDX, XPS, TOF-SIMS, and variable-angle FTIR. Advancing aqueous contact angle measurements were used to evaluate the nanofiber surface wettability. PMID- 17402762 TI - 129Xe NMR spectroscopy study of porous cyanometallates. AB - Zinc and cadmium hexacyanocobaltates(III) were prepared, and their porous networks were explored using 129Xe spectroscopy. The crystal structures of these two compounds are representative of porous hexacyanometallates, cubic (Fm-3m) for cadmium and rhombohedral (R-3c) for zinc. In the cubic structure, the porosity is related to systematic vacancies created from the elemental building block (i.e., the hexacyanometallate anion), whereas the rhombohedral (R-3c) structure is free of vacant sites but has tetrahedral coordination for the zinc atom, which leads to relatively large ellipsoidal pores communicated by elliptical windows. According to the Xe adsorption isotherms, these porous frameworks were found to be accessible to the Xe atom. The structure of the higher electric field gradient at the pore surface (Fm-3m) appears and is accompanied by a stronger guest-host interaction for the Xe atoms and a higher capacity for Xe sorption. For cadmium, the 129Xe NMR signal is typical of isotropic movement for the Xe atom, indicating that it remains trapped within a spherical cavity. From spectra recorded for different amounts of adsorbed Xe, the cavity diameter was estimated. For the zinc complex, 129Xe NMR spectra are asymmetric because of the Xe atom movement within an elongated cavity. The line-shape asymmetry changes when the Xe loading within the porous framework increases, which was ascribed to Xe-Xe interactions through the cavity windows. The Xe adsorption revealed additional structural information for the studied materials. PMID- 17402763 TI - Fatty acids influence "solid" phase formation in models of stratum corneum intercellular membranes. AB - Stacked intercellular lipid membranes in the uppermost epidermal layer, the stratum corneum (SC), are responsible for skin's barrier function. These membranes are unique in composition, the major lipids being ceramides (Cer), cholesterol, and free fatty acids (FFA) in approximately equimolar proportions. Notably, SC lipids include chains much longer than those of most biological membranes. Previously we showed that Cer's small hydrophilic headgroup enabled SC model membranes composed of bovine brain ceramide (BBCer), cholesterol, and palmitic acid in equimolar proportion to solidify at pH 5.2. In order to determine the influence of FFA chain length on the phase behavior of such membranes, we used 2H NMR and FT-IR to study BBCer/cholesterol/FFA dispersions containing linear saturated FFA 14-22 carbons long. Independent of chain length, the solid phase dominated the FFA spectrum at physiological temperature. Upon heating, each dispersion underwent phase transitions to a liquid crystalline phase (only weakly evident for the membrane containing FFA-C22) and then to an isotropic phase. The phase behavior, the lipid mixing properties, and the transition temperatures are shown to depend strongly on FFA chain length. A distribution of FFA chain lengths is found in the SC and could be required for the coexistence of a proportion of solid lipids with some more fluid domains, which is known to be necessary for normal skin barrier function. PMID- 17402764 TI - A disjoining pressure study of foam films stabilized by mixtures of nonionic and ionic surfactants. AB - Studying the disjoining pressure Pi as a function of the film thickness h (Pi-h curves) of foam films stabilized by ionic and nonionic surfactants, one finds that the surface charge density q0 of films stabilized by ionic surfactants increases with increasing surfactant concentration, while the opposite holds true for nonionic surfactants. Thus, it should be possible to tune the surface charge density with mixtures of nonionic and ionic surfactants. To address this question, we studied foam films stabilized by aqueous solutions of surfactant mixtures. The mixtures consisted of the nonionic beta-dodecylmaltoside (beta C12G2) and the cationic dodecyl trimethylammonium bromide (C12TAB) with mixing ratios of beta-C12G2/C12TAB = 1:0, 50:1, 1:1, 1:50, 0:1. The addition of small amounts of C12TAB to beta-C12G2 first neutralizes the negative surface charge of the beta-C12G2 films and finally leads to a charge reversal from negatively to positively charged surfaces. On the other hand, by adding small amounts of beta C12G2 to C12TAB, one observes the formation of stable CBFs which was also observed for the pure C12TAB. However, in contrast to the pure C12TAB, the resulting Pi-h curves for the mixtures cannot be described with the Derjaguin Landau-Verwey-Overbeek (DLVO) theory; the slope of the curves is too steep, and it barely changes with changing electrolyte concentration. A possible explanation for this observation will be given and discussed. PMID- 17402765 TI - Supramolecular polymers based on the quadruplex formation of ditopic guanosine macromonomers in nonaqueous media. AB - The formation of supramolecular polymeric aggregates with a molecular mass of 100 kDa in a nonaqueous solution from a telechelic dimer of isopropylidene guanosine in the presence of K(+) ions is reported. The possible structure of macromonomers resulting from the development of G4 quartets was deduced from DOSY NMR, circular dichroism spectra, and dynamic light scattering measurements. PMID- 17402766 TI - Light backscattering as an indirect method for detecting emulsion inversion. AB - Many phenomena take place during different types of emulsion inversions, particularly a change in interface curvature and drop size, which could be detected by backward light scattering. Monitoring the backscattering signal allows us to detect the emulsion inversion in three main cases, one transitional and two catastrophic types. The backscattering data could give some clue as to emulsion morphology, which is not available from conductivity measurements. PMID- 17402767 TI - Spontaneous two-dimensional spherical colloidal structures. AB - Two-dimensional spherical crystalline colloidal structures are formed at the interface between water and oil as the result of spontaneous emulsification and colloidal self-assembly. When water droplets are introduced in oil containing a lipophilic surfactant, smaller water droplets of uniform size are spontaneously produced at the spherical interface. Initially of submicrometer size, the small droplets at the interface self-assemble, forming ordered structures, and grow uniformly with time until they reach a size of a few micrometers, maintaining the crystalline structure. PMID- 17402768 TI - Contact angles of submillimeter particles: connecting wettability to nanoscale surface topography. AB - A method is presented for the measurement of contact angles of particles in the size range 100-1000 microm, using an optical microscope. This method is used to characterize the wettability of polyethylene particles produced in a gas-phase polymerization process. The as-polymerized particles are shown to be significantly more hydrophobic than melt-processed polyethylene materials. The surface structure of the particles is examined with scanning electron microscopy, and the increased hydrophobicity of the as-polymerized particles is attributed to the nanoscale texture of the particle surfaces. PMID- 17402769 TI - Mining phosphopeptide signals in liquid chromatography-mass spectrometry data for protein phosphorylation analysis. AB - Protein phosphorylation is a key post-translational modification that governs biological processes. Despite the fact that a number of analytical strategies have been exploited for the characterization of protein phosphorylation, the identification of protein phosphorylation sites is still challenging. We proposed here an alternative approach to mine phosphopeptide signals generated from a mixture of proteins when liquid chromatography-tandem mass spectrometry (LC MS/MS) analysis is involved. The approach combined dephosphorylation reaction, accurate mass measurements from a quadrupole/time-of-flight mass spectrometer, and a computing algorithm to differentiate possible phosphopeptide signals obtained from the LC-MS analyses by taking advantage of the mass shift generated by alkaline phosphatase treatment. The retention times and m/z values of these selected LC-MS signals were used to facilitate subsequent LC-MS/MS experiments for phosphorylation site determination. Unlike commonly used neutral loss scan experiments for phosphopeptide detection, this strategy may not bias against tyrosine-phosphorylated peptides. We have demonstrated the applicability of this strategy to sequence more, in comparison with conventional data-dependent LC MS/MS experiments, phosphopeptides in a mixture of alpha- and beta-caseins. The analytical scheme was applied to characterize the nasopharyngeal carcinoma (NPC) cellular phosphoproteome and yielded 221 distinct phosphorylation sites. Our data presented in this paper demonstrated the merits of computation in mining phosphopeptide signals from a complex mass spectrometric data set. PMID- 17402770 TI - Determining compositional profiles within conducting polymer films following reaction with vapor phase reagents. AB - A combination of XPS, reflectance infrared spectroscopy, and neutron reflectivity measurements has been used to probe the spatial and global extents to which carboxylic acid motifs in electrodeposited conducting polymer films can be functionalized by reaction with vapor phase reagents (a carbodiimide together with trifluoroethanol) with the goal of controlling hydrophobicity. Across a range of polymer deposition and reaction temperatures, neutron reflectivity showed that, surprisingly, functionalization of the polymer matrix at depths >5 10 nm into the polymer film was always significantly lower than at the exposed surface. The most efficient functionalization was found to occur when a low density polymer matrix was prepared by elution of motifs cleaved from the polymer by base hydrolysis. Finally, when trifluoroethanol functionalization was performed, the macroscopic property of hydrophobicity was related to the surface, internal microstructure, and composition of the reacted films as elucidated by the above combination of probes. PMID- 17402771 TI - A molecular dynamics simulation study of C60 fullerenes inside a dimyristoylphosphatidylcholine lipid bilayer. AB - We have carried out atomistic molecular dynamics simulations of C60 fullerenes inside a dimyristoylphosphatidylcholine lipid bilayer and an alkane melt. Simulations reveal that the preferred position of a single C60 fullerene is about 6-7 A off of the center plane, allowing the fullerene to take advantage of strong dispersion interactions with denser regions of the bilayer. Further displacement (>8 A) of the fullerene away from the center plane results in a rapid increase in free energy likely due to distortion of the lipid head group layer. The effective interaction between fullerenes (direct interaction plus environment (bilayer) induced interaction), measured as the potential of mean force (POMF) between two fullerenes as a function of their separation, was found to be significantly less attractive in the lipid bilayer than in an alkane melt of the same molecular weight as the lipid tails. Only part of this difference can be accounted for by the more favorable interaction of the fullerene with the relatively denser bilayer. Additionally, our POMF studies indicate that the bilayer is less able to accommodate the larger aggregated fullerene pair than isolated single fullerenes, again likely due to distortion of the bilayer structure. The implications of these effects on aggregation of fullerenes within lipid bilayer are considered. PMID- 17402772 TI - Amorphous iron(III) oxide--a review. AB - The syntheses of amorphous Fe(2)O(3) nanoparticles of varying size and morphology, their magnetic properties, crystallization mechanism, and applications are reviewed herein. The synthetic routes are classified according to the nature of the sample (powders, nanocomposites, films, coated particles). The contributions of various experimental techniques to the characterization of an amorphous Fe(2)O(3) phase are considered in this review, including some key experimental markers, allowing its distinction from nanocrystalline "X-ray amorphous" polymorphs (maghemite, hematite). We discuss the thermally induced crystallization mechanisms depending on transformation temperature, atmosphere, and the size of the amorphous particles that predetermine the structure of the primarily formed crystalline polymorph. The controversial description of the magnetic behavior, including an interpretation of the low-temperature and in field Mossbauer spectra, is analyzed. PMID- 17402773 TI - Electronic and molecular structures of trigonal truxene-core systems conjugated to peripheral fluorene branches. Spectroscopic and theoretical study. AB - An analysis is performed on the molecular and electronic features in a series of trigonal molecules constituted by a central truxene core which is ramified with three oligofluorene moieties of different lengths. Arms and core are studied independently and upon threefold unification. Special emphasis is paid to the modulation of the conjugational properties in relation to substitution, molecular dimension, ring aromaticity, intermolecular forces, oxidation state, etc. Raman and optical absorption/emission spectroscopies in conjunction with computational theoretical results are combined for this purpose. The evolution of some key intensity ratios in the Raman spectra (i.e., I(1300)/I(1235)) is followed as an indication of electronic interaction between the core and the branches. The changes of the electronic delocalization upon solvation, with varying temperature in the solid state, with the nature of the aromatic unit (bithiophene/fluorene) or after electrochemical oxidation are interpreted. The modulation of the optical properties on the basis of the structure and energetics of the orbital around the gap is also addressed. Density functional theory was used to assign the vibrational and electronic spectra. PMID- 17402774 TI - Complexes of polyelectrolyte-neutral double hydrophilic block copolymers with oppositely charged surfactant and polyelectrolyte. AB - Complexes between sodium (sulfamate-carboxylate)isoprene/ethylene oxide double hydrophilic diblock copolymers (SCIEO) and dodecyltrimethylammonium bromide (DTMAB), as well as quaternized poly(2-vinylpyridine) (QP2VP), were studied in aqueous solutions, at pH 7. The complexes are formed due to electrostatic interactions between the anionic groups of the polyelectrolyte block of the copolymers and the cationic groups of the surfactant or the homopolyelectrolyte. The structure of the complexes was investigated as a function of the mixing ratio of the two components in solution and ionic strength by static, dynamic, and electrophoretic light scattering, atomic force microscopy, and fluorescence spectroscopy. The mass and size of the complexes depend on the mixing ratio between the components. A transition from intrachain to an interchain association was observed for block copolymer/ surfactant complexes. SCIEO/QP2VP complexes were found to respond to increasing concentrations of added salt. Spherical and ellipsoid shaped complexes with a core-shell micellar like structure were formed in the systems studied. PMID- 17402775 TI - Effect of an anionic surfactant on the complexation of some nonionic polymers with iodine in aqueous media. AB - Complexation of some water soluble nonionic polymers, namely, polyvinylalcohol (PVA), polyvinylpyrrolidone (PVP), and hydroxypropyl cellulose (HPC), with iodine has been studied in aqueous and aqueous sodiumdodecylsulfate (SDS) solution. While the complexation was indicated by a red shift of the tri-iodide band in case of PVP or HPC, the PVA-iodine complex showed its characteristic band around 500 nm. It was observed for the first time that presence of SDS led to complete break down of the PVA-iodine complex and its characteristic blue color. The presence of monomers of SDS, however, appeared to favor the formation of the iodine complex with PVP or HPC. Addition of n-propanol, which is known to prevent the formation of gels or microgels in polymer solutions, was found to enhance the polymer-iodine complex. Gels of pure HPC and HPC with iodine both in presence and absence of SDS have been prepared and studied. PMID- 17402776 TI - Interfacial electron transfer in metal cyanide-sensitized TiO2 nanoparticles. AB - Electroabsorption (Stark) spectroscopy has been used to study the charge-transfer absorption from a transition-metal-cyanide complex to a TiO2 nanoparticle. Transition-metal cyanide/TiO2(particle) systems were synthesized using FeII(CN)(6)4-, RuII(CN)6(4-), MoIV(CN)(8)4-, and WIV(CN)8(4-). On formation of the M(CN)n4-/TiO2(particle) system, a new metal-to-particle charge-transfer (MPCT) absorption band is observed in the 390-480 nm region. Analysis of the absorption spectra suggests that the TiO2 level involved in the MPCT transition resides at significantly higher energy than the bottom of the conduction band and that the electronic coupling between the two metal centers is the dominant factor determining the position of the MPCT band maximum. The average charge-transfer distances determined by Stark spectra range from 4.1-4.7 A. The observation of relatively short charge-transfer distances leads to the conclusion that the MPCT absorption is from the transition-metal cyanide center to a level that is localized on the Ti atom bound to a nitrogen end of the [O2Ti-N-C-M(CN)x] system. The electronic coupling, Hab, calculated for a two state model is similar to values observed in dinuclear metal complexes. PMID- 17402777 TI - Ion distribution in polyelectrolyte multilayers with standing-wave X-ray fluorescence. AB - Absolute ion concentration and its profile across polyelectrolyte multilayer films were studied. The films were prepared by alternating adsorption of polyanions and polycations from aqueous solution. Standing-wave X-ray fluorescence was used to map the ion profile. The well-studied multilayer system PSS/PAH was investigated, and bromide ions were used as probe entities. The results show that the sign of the charge of the outermost layer and the washing procedure after finishing the preparation have a decisive effect on the ion concentration and the ion profile. Multilayers with PSS as the outermost layer contain fewer bromide ions than the PAH-terminated multilayers. Exposure to water washes the ions out, but even after 6 h of washing, not all of the bromide ions had been removed. PMID- 17402778 TI - Structure and dynamics of sulfate ion in aqueous solution--an ab initio QMCF MD simulation and large angle X-ray scattering study. AB - The hydrated sulfate ion has been characterized in aqueous solution in structural and dynamic aspects using ab initio quantum mechanical charge field (QMCF) molecular dynamics (MD) simulation and large angle X-ray scattering (LAXS) methods. The LAXS data show an average coordination number of the sulfate ion of up to 12 water molecules bound through hydrogen bonding, while the QMCF MD simulation displays a wide range of coordination numbers between 8 and 14 with an average value of approximately 11. The Os...Ow distance cannot be distinguished from the Ow...Ow distance in the LAXS experiment; the weighted mean O...O distance is 2.880(10) A. In the simulation, the Os...Ow and Ow...Ow distances are found to be very similar, namely, 2.86 and 2.84 A, respectively. The S-Os bond and S...Ow distance have been determined by the LAXS experiment as 1.495(6) and 3.61(2) A, respectively, indicating an average nearly tetrahedral S-Os...Ow angle. The approximately 5% deviations of simulation distances (1.47 and 3.82 A) from the experimental ones can probably be ascribed to the neglect of correlation energy in the quantum mechanical method. The mean residence time of water ligands at O atoms, 2.57 ps, is longer than that in pure water, 1.7 ps, characterizing the sulfate ion as a weak structure maker. PMID- 17402779 TI - Stochastic simulations of the cytochrome P450 catalytic cycle. AB - Cytochrome P450 enzymes involve a complex reaction cycle which has been described, for the first time, by a stochastic simulation of the system in the present work. A series of models are developed for a basic catalytic cycle, employing a set of microscopic rate constants for the oxidation of p alkoxyacylanilides catalyzed by the cytochrome P450 1A2. By analyzing the effects of low concentrations of enzyme and substrate on the system, and the dependence of the system on several rate constants, it is discovered that the system evolves along relatively stable patterns from its initial state, as indicated from different runs of simulations. Strong fluctuations appear at the entrance and exit of the pathway, with very weak fluctuations in the middle sections of the cycle. Although noises are apparent when the reactant populations are very low, basically, the fundamental feature of the P450 cycle based on a microscopic view is that it is deterministic in nature. Meanwhile, the mathematical models we developed are qualitatively validated by a comparison with those experimental results of the P450 cycle. The findings of this work will be helpful for a further deeper understanding of the catalytic mechanism of cytochrome P450 enzymes. PMID- 17402780 TI - Rheology of gastric mucin exhibits a pH-dependent sol-gel transition. AB - Gastric mucin, a high molecular weight glycoprotein, is responsible for providing the gel-forming properties and protective function of the gastric mucus layer. Bulk rheology measurements in the linear viscoelastic regime show that gastric mucin undergoes a pH-dependent sol-gel transition from a viscoelastic solution at neutral pH to a soft viscoelastic gel in acidic conditions, with the transition occurring near pH 4. In addition to pH-dependent gelation behavior in this system, further rheological studies under nonlinear deformations reveal shear thinning and an apparent yield stress in this material which are also highly influenced by pH. PMID- 17402781 TI - Cascade polymeric MRI contrast media derived from poly(ethylene glycol) cores: initial syntheses and characterizations. AB - Diagnostic contrast media for magnetic resonance imaging (MRI) are often applied to enhance the signal of blood allowing for quantitative definition of vascular functional characteristics including tissue blood volume, flow, and leakiness. Well-tolerated and safe macromolecular formulations are currently being sought that remain in the blood for a relatively long period and that leak selectively from diseased vessels, particularly cancer vessels. We synthesized a new class of macromolecular, water-soluble MRI contrast media by introducing two diverging polylysine cascade amplifiers at each end of a poly(ethylene glycol) (PEG) backbone, followed by substitution of terminal lysine amino groups with Gd-DTPA chelates. Four candidate PEG cascade conjugates are reported here, PEG3400-Gen4 (Gd-DTPA)8, PEG6000-Gen4-(Gd-DTPA)8, PEG12000-Gen4-(Gd-DTPA)8, and PEG3400-Gen5 (Gd-DTPA)13 with descriptions of their basic physical, biological, and kinetic properties, including real and effective molecular sizes, proton T1 relaxivities in water and plasma, partition coefficients, osmolalities, chelate stability, stability in plasma, stability to autoclaving, certain in vivo pharmacokinetics (blood half-life, blood clearance, volume of distribution), and whole body elimination profiles in normal rodents. These candidate PEG-core cascade MRI contrast media showed a range of effective molecular sizes similar to proteins weighing 74-132 kDa, although their actual molecular weights were much smaller, 12-20 kDa. All compounds exhibited a narrow range of size dispersity and relatively high T1 relaxivities (approximately 3 times the value for unconjugated Gd-DTPA at 2 T and 37 degrees C). Representative compounds also showed a high degree of hydrophilicity, stability in solution buffer and plasma, and lack of binding to proteins. The two candidate compounds with the largest effective molecular sizes, PEG12000-Gen4-(Gd-DTPA)8 and PEG3400-Gen5-(Gd-DTPA)13, had longer blood half-lives, 36 and 73 min, respectively (monoexponential kinetics for both), and showed strong, prolonged MRI enhancement of vessels. Results also indicate that in vivo pharmacokinetics and bodily elimination profiles can be adjusted by the selection of molecular size for the PEG core and the selection of the amplification degree of the cascade polylysine clusters. The initially evaluated compounds from this new class of contrast media show acceptable, desirable characteristics in many, but not all, respects. Further efforts are directed toward candidate macromolecules having higher thermodynamic stability, higher degree of substitution by gadolinium chelates, and more rapid bodily elimination. PMID- 17402782 TI - Macroscopic fibers self-assembled from recombinant miniature spider silk proteins. AB - Strength, elasticity, and biocompatibility make spider silk an attractive resource for the production of artificial biomaterials. Spider silk proteins, spidroins, contain hundreds of repeated poly alanine/glycine-rich blocks and are difficult to produce recombinantly in soluble form. Most previous attempts to produce artificial spider silk fibers have included solubilization steps in nonphysiological solvents. It is here demonstrated that a miniature spidroin from a protein in dragline silk of Euprosthenops australis can be produced in a soluble form in Escherichia coli when fused to a highly soluble protein partner. Although this miniature spidroin contains only four poly alanine/glycine-rich blocks followed by a C-terminal non-repetitive domain, meter-long fibers are spontaneously formed after proteolytic release of the fusion partner. The structure of the fibers is similar to that of dragline silks, and although self assembled from recombinant proteins they are as strong as fibers spun from redissolved silk. Moreover, the fibers appear to be biocompatible because human tissue culture cells can grow on and attach to the fibers. These findings enable controlled production of high-performance biofibers at large scale under physiological conditions. PMID- 17402783 TI - Constituents of Oxandra cf. xylopioides with anti-inflammatory activity. AB - A new triterpenoid (1) derived from 24-methylcycloartanol was isolated from the leaves of Oxandra cf. xylopioides. An unusual structure of the new compound was assigned as 1, for which the name berenjenol is proposed, on the basis of the spectroscopic data of the natural product and of its derivatives 2 and 3. The leaves also afforded the known monoterpene isoespintanol (4). Compounds 1 and 4 significantly reduced the paw edema induced by carrageenan by 64% and 43%, at 3 h, respectively. Moreover, 4 reduced IL-1 beta production by 72% at 100 microM and reduced IL-1 beta mRNA synthesis. PMID- 17402784 TI - Ruthenium-catalyzed cyclization of 2-alkyl-1-ethynylbenzenes via a 1,5-hydrogen shift of ruthenium-vinylidene intermediates. AB - Catalytic cyclization of 2-alkyl-1-ethynylbenzene derivatives was implemented by TpRuPPh3(CH3CN)2PF6 (10 mol %) in hot toluene (105 degrees C, 36-100 h) to form 1 substituted-1H-indene and 1-indanone products; such cyclizations proceeded more efficiently for substrates bearing electron-rich benzenes. We propose that the cyclization mechanism involves a 1,5-hydrogen shift of initial metal-vinylidene intermediate. PMID- 17402785 TI - Suzuki cross-coupling on enantiomerically pure epoxides: efficient synthesis of diverse, modular amino alcohols from single enantiopure precursors. AB - Suzuki arylation of enantiopure (4-bromophenyl)- or (3,5-dibromophenyl)glycidyl ethers has been achieved in toluene under Buchwald conditions [ArB(OH)2 (1.1 equiv), Cs2CO3 (2 equiv), Pd2(dba)3.C6H6 (1 mol %), S-Phos (4 mol %), toluene, 100 degrees C] allowing for the formation of modular arylglycidyl ethers not directly available in enantiopure form by epoxidation routes. These bulky ethers, when submitted to regioselective and stereospecific ring opening with ammonia [aq NH3, LiClO4 (1 equiv), THF, microwave irradiation (80 W), 125 degrees C] in a sealed tube, provide access to novel enantiopure beta-amino alcohols which, in turn, provide an easy access to structurally complex C2 symmetrical bisoxazolines. PMID- 17402786 TI - Diastereoselective synthesis of alpha-methyl and alpha-hydroxy-beta-amino acids via 4-substituted-1,3-oxazinan-6-ones. AB - 1,3-oxazinan-6-ones have been utilized in a series of enolate reactions to produce 5-hydroxy and 5-alkyl-4-substituted-1,3-oxazinan-6-ones with excellent trans diastereoselectivity. Highlighting the versatility of the oxazinanone, a number of transformations were performed to produce a variety of protected N-H and N-methyl alpha-hydroxy- and alpha-methyl-beta-amino acids. PMID- 17402787 TI - Remote hydroxylation of methyl groups by regioselective cyclopalladation. Partial synthesis of hyptatic acid-A. AB - Hyptatic acid-A (32), a 2alpha,3beta,24-trihydroxyolean-12-en-28-oic acid, previously isolated from Hyptis capitata, was obtained from maslinic acid (2). The regioselective cyclopalladation of the axial methyl group on C-4 of maslinic acid afforded the C-24 hydroxymethylene group due to the presence of a C-2-OR substituent. Nevertheless, hederagenin (7) (23-hydroxy derivative) was formed when this oxygenated group was not present. PMID- 17402788 TI - Synthetic studies on macrolactin A: construction of C4-C24 fragment. AB - The preparation of the C4-C24 fragment of the macrolactin A is described. The adopted synthetic strategy involves two isomerizations to selectively construct the (8E,10Z) and (16E,18E) dienes, using a sequential Claisen rearrangement/allene isomerization and a Ph3P-catalyzed isomerization of an yne one, respectively. PMID- 17402789 TI - Facile electrochemical characterization of core/shell nanoparticles. Ag core/Ag(2)O shell structures. AB - We report in this paper a facile approach for the formation and electrochemical characterization of silver-silver oxide core-shell nanoparticles (NPs). Thus, thermal treatment at temperatures between 200 and 360 degrees C of Ag NP, in the gas phase or in an organic solvent, has been used to achieve the formation Ag@Ag2O NP. The evidence of formation of such a core-shell structure was obtained by cyclic voltammetry using a Nafion modified electrode (where Nafion containing carbon particles is used as the matrix to encapsulate the core-shell NP). Initial positive scans measure free Ag. Initial negative scans measure Ag2O, with the following positive scan, compared to the initial one, providing a measure of "trapped" or core Ag. The results presented demonstrate the utility of this approach in characterizing core-shell structures, like Ag@Ag2O, which could be extended to other core-shell forms, such as bimetallic core-shell NP. PMID- 17402790 TI - HER-2-positive breast cancer: hope beyond trastuzumab. AB - Therapeutic antibodies have shown great promise as targeted agents in the treatment of patients with cancer. Trastuzumab, a humanized monoclonal antibody targeting human epidermal growth factor receptor-2 (HER-2), is of special importance in breast cancers overexpressing HER-2. Such rationally designed substances bind to cancer cells expressing the targeted antigen and, by various mechanisms, lead to tumor cell degradation. Only one-third of patients, however, initially respond to trastuzumab monotherapy and the majority of initial responders demonstrate disease progression within 1 year of treatment initiation. Therefore, alternative compounds targeting the HER-2 receptor or downstream signaling pathways are of great importance. Lapatinib is a tyrosine kinase inhibitor, blocking tryosine kinase domains of both epidermal growth factor receptor and HER-2. This substance holds promise for the treatment of cancer after trastuzumab failure, and might be active in cerebral metastases. Other strategies in trastuzumab-resistant disease include bispecific antibodies (which bind to HER-2 and Fc receptors, thereby directing immune cells towards the tumor), the combination of antibodies, or targeting tumor vessel growth by blocking vascular endothelial growth factor (VEGF) or VEGF receptors. Heat shock protein 90, a chaperone protein that controls the folding of HER-2, also represents a potential target. Multi-targeted kinase inhibitors such as sunitinib or sortenib are already established in renal cell cancer. These compounds are currently being evaluated in breast cancer and might represent interesting options both in HER-2-positive and -negative disease. In conclusion, trastuzumab remains the gold standard in HER-2-positive breast cancer therapy. However, in trastuzumab-resistant disease, new strategies and compounds are currently under evaluation. PMID- 17402791 TI - Ethics and embryonic stem cell research: altered nuclear transfer as a way forward. AB - Ethical controversy in stem cell research arises because current methods to produce embryonic stem cell lines require the destruction of living human embryos. For this reason, there is increasing interest in developing alternative, non-embryonic sources of pluripotent stem cells. This effort is especially important in the US due to the prevailing policy against federal funding of embryo-destructive research. Altered nuclear transfer (ANT) is one of several potential methods to develop alternative sources of pluripotent stem cells. This approach employs the technique of somatic cell nuclear transfer, but the somatic cell nucleus or egg cytoplasm (or both) are first altered before the somatic cell nucleus is transferred into the oocyte. This alteration precludes the coordinated organization and developmental potential that is necessary for the resulting biological entity to be an embryo, but it still allows the entity to generate pluripotent stem cells. Proof-of-principle for one variant of ANT has been established in mice by silencing the functional expression of the gene Cdx2 in the somatic cell nucleus prior to its transfer into an enucleated egg. From the resulting non-embryonic laboratory construct, fully functional pluripotent stem cells were procured. Other more recent studies have suggested the possibility of achieving the same results by preemptively silencing maternally derived Cdx2 messenger RNA in the egg before the act of nuclear transfer. The procedure would produce the equivalent of a tissue culture of pluripotent stem cells. In contrast to the use of embryos 'left over' from clinical in vitro fertilization, ANT could produce pluripotent stem cell lines with an unlimited range of specifically selected and controlled genotypes. Such flexibility would greatly facilitate the study of disease, drug development, and toxicology testing, and may allow the production of therapeutically useful pluripotent stem cells that are immune compatible. If developed to the point of scientific reliability, ANT would be a valuable research tool for the study of other aspects of cell development and differentiation, including gene expression patterns, imprinting, and cell-cell signaling. ANT would also help to clarify definitions and boundaries that distinguish true organisms from 'biological artifacts' and, thereby, provide moral precedent to guide future progress in developmental biology. PMID- 17402792 TI - Targeting the phosphatidylinositol 3-kinase pathway in airway smooth muscle: rationale and promise. AB - The phosphatidylinositol 3-kinase (PI3K) signaling pathway plays a critical role in regulating cell growth, proliferation, survival, and motility. Structural alterations, e.g. airway remodeling, in asthma and chronic obstructive pulmonary disease (COPD) are associated with increased airway smooth muscle (ASM) cell growth and proliferation due to the frequent stimulation of ASM by inflammatory mediators, contractile agonists, and growth factors. The critical role of the PI3K signaling pathway in regulating ASM cell growth and proliferation is well established. However, recent discovery of the tumor suppressor proteins tuberous sclerosis complex 1 (TSC1) and TSC2, also known as hamartin and tuberin, as downstream effectors of PI3K and upstream regulators of the mammalian target of rapamycin (mTOR) and S6 kinase 1(S6K1) shed a new light on the PI3K signaling cascade in regulating cell growth and proliferation. The activity of TSC1/TSC2 is regulated by growth factors, nutrients, and energy; thus, TSC1/TSC2 serves as a signaling module for protein translational regulation, cell cycle progression, and cell size, which are key events controlling cell growth and proliferation. This article highlights the potential contribution of the PI3K-TSC1/TSC2 mTOR/S6K1 pathway in smooth muscle remodeling. Pharmacologic manipulation of this signaling pathway could have a major impact on treatment of asthma and COPD. PMID- 17402793 TI - MicroRNAs in the search for understanding human diseases. AB - MiroRNAs (miRNAs) are double-stranded, noncoding RNA molecules (with an average size of 22bp) that serve as post-transcriptional regulators of gene expression in higher eukaryotes. miRNAs play an important role in development and other cellular processes by hybridizing with complementary target mRNA transcripts, preventing their translation and thereby destabilizing the target transcripts. Though hundreds of miRNAs have been discovered in a variety of organisms, little is known about their cellular function. They have been implicated in the regulation of developmental timing and pattern formation, restriction of differentiation potential, regulation of insulin secretion, resistance to viral infection, and in genomic rearrangements associated with carcinogenesis or other genetic disorders, such as fragile X syndrome. Recent evidence suggests that the number of unique miRNA genes in humans exceeds 1000, and may be as high as 20,000. It is estimated that 20-30% of all human mRNAs are miRNA targets. During the last few years, special attention has been given to miRNAs as candidate drug targets for cancer, diabetes mellitus, obesity, and viral diseases. PMID- 17402794 TI - Immunoglobulin treatment for primary antibody deficiencies: advantages of the subcutaneous route. AB - Adults and children with primary antibody deficiencies are prone to bacterial infections affecting the respiratory tract and gastrointestinal canal. To prevent or alleviate infections, replacement therapy with IgG is needed, usually on a lifelong basis. The IgG can be administered intramuscularly, intravenously, or subcutaneously. Subcutaneous IgG (SCIG) therapy, using small portable pumps for once-per-week self infusions, has shown many advantages compared with the two other routes of administration. This review highlights findings from international studies and demonstrates that: (i) SCIG therapy is safe, with very few adverse effects; (ii) the therapy can be used for patients with previous adverse effects to intravenous administration of IgG; (iii) the therapy leads to high serum IgG levels and good protection against infections; (iv) the therapy facilitates home therapy, as the infusion technique is easy for children, adults and elderly people to learn and there is no need for venous access; (v) SCIG home therapy leads to significantly improved life situations for the patients; (vi) the SCIG home therapy regimen in particular reduces the costs of treatment. PMID- 17402795 TI - Classification and predictive modeling of liver X receptor response elements. AB - BACKGROUND: The liver X receptor (LXR), a transcription factor that forms a heterodimer with the retinoid X receptor, plays a key role in the transcriptional regulation of many important genes implicated in prevalent metabolic diseases. In spite of numerous studies, a complete list of LXR direct target genes remains elusive. To complement experimental approaches, computational prediction can be used to help build such a list because all LXR target genes are expected to carry the response elements (LXREs) in their promoter or enhancer regions. In practice, however, such a prediction has been hampered by the inaccuracies of currently available predictive models of LXREs. We report on a novel computational application for the highly accurate prediction of LXREs in DNA sequences. METHODS: We first conducted a comprehensive review of experimentally determined LXR target genes and collected all known LXREs. Subsequently, all such sites were classified using various computational methods based on sequence similarity to identify multiple subtypes. A library of Hidden Markov Models (LXRE.HMM) was developed to represent all subtypes and to enable the promoter scanning of LXR target genes. RESULTS AND CONCLUSION: Our model outperformed the widely used LXRE model in MatInspector in identifying the LXREs for all known LXR direct target genes at the experimentally verified positions. As a result, this new approach will make the genomewide prediction of LXR target genes feasible. PMID- 17402796 TI - Adalimumab: in Crohn's disease. AB - Adalimumab is a subcutaneously administered, recombinant, human IgG1 monoclonal antibody specific for human tumor necrosis factor (TNF). The clinical efficacy and safety of adalimumab in patients with moderate to severe Crohn's disease has been demonstrated in four pivotal, randomized, double-blind trials (CLASSIC-I, GAIN, CHARM, and CLASSIC-II) that included a total of >1400 patients. In the CLASSIC-I trial, adalimumab was significantly more effective than placebo for induction of remission in patients who had not previously received anti-TNF therapy. Adalimumab was also more effective than placebo for induction of remission in the GAIN study in patients who had either lost responsiveness or developed intolerance to infliximab. The CHARM trial showed that, among patients who responded to open-label adalimumab induction, maintenance therapy with adalimumab 40 mg weekly or every other week for up to 1 year was associated with significantly greater remission rates than placebo at weeks 26 and 56. In addition, significantly more adalimumab than placebo recipients achieved corticosteroid-free remission and had complete fistula closure. In CLASSIC-II, an extension of the CLASSIC-I trial, patients who were in remission after a short course of adalimumab and were randomized to receive up to 1 year's treatment with adalimumab 40 mg weekly or every other week were significantly more likely to remain in remission than those randomized to receive placebo. In general, the tolerability profile of adalimumab in patients with Crohn's disease was similar to that in patients with rheumatoid arthritis or other approved indications. PMID- 17402800 TI - Panitumumab in the treatment of metastatic colorectal cancer: profile report. PMID- 17402801 TI - Letter from the Commissioner's Office at the US FDA. PMID- 17402802 TI - Measuring patient-reported outcomes in solid organ transplant recipients: an overview of instruments developed to date. AB - Improvements in organ preservation methods, immunosuppressive regimens and general post-transplant care have resulted in an increased life expectancy and a continually decreasing morbidity after solid organ transplantation. As attention gradually moves towards improving subjective patient outcomes, the use of patient reported outcome measures (PROs) thus becomes increasingly important in post transplant patient management. This paper provides a brief systematic overview of the transplant-specific PRO measures, and their psychometric properties, developed and used in solid organ transplant recipients to date. PRO measures may focus on or encompass different aspects of life relevant to organ transplant patients: overall quality of life (QOL), physical, psychological and social functioning, and adherence. Overall QOL can be measured using transplant-specific or generic QOL instruments, or a combination of both. In general, very little information is available on the psychometric properties of PRO measures, and there is no gold standard for PRO measurement. Transplant-specific and generic PRO instruments are complementary. Generic instruments will continue to be important for economic evaluations, but transplant-specific instruments may be more useful for patient management purposes, as they are generally more sensitive to small but clinically relevant changes in outcomes in transplant populations. PMID- 17402803 TI - The economic implications of glaucoma: a literature review. AB - Glaucoma is a common ophthalmic condition, often associated with elevated intraocular pressure (IOP). It affects >2 million people in the US, and the incidence is expected to exceed 3 million by 2020. However, relatively little is known about the cost of glaucoma compared with costs for other eye conditions. This comprehensive report reviews published literature on costs and cost effectiveness of treatments for glaucoma. Cost-of-illness studies in glaucoma focus on direct medical costs and generally exclude indirect costs. In general, increased costs are associated with increased severity or lack of control over IOP and the distribution of costs (e.g. medication vs procedures) varies with severity. A large number of studies have evaluated the cost of glaucoma medications, assessing the number of drops per bottle and associated cost per drop or per treatment dose. These studies have limited usefulness as they generally evaluate unit medication costs without including differential effectiveness or adverse effects associated with various therapies, and thus provide only one component of real-world costs for glaucoma. Broader comparative cost studies, mainly adopting a cost-minimisation approach, have evaluated the impact of differing treatments and management strategies on all types of medical care resource utilisation and associated costs, but a variety of metrics for success makes interpretation challenging. Studies have generally found beta2 adrenoceptor antagonists to be associated with greater healthcare costs than newer therapies. Among newer treatments such as prostaglandin analogues, no specific treatment has demonstrated a clear cost advantage over other treatments. A number of studies have modelled hypothetical cohorts of glaucoma patients through courses of therapy, projecting costs, outcomes and cost effectiveness. A majority of these cost-effectiveness models compare one of the newer prostaglandin analogues with older medications or with one another. Existing studies suggest that bimatoprost may be more cost effective than other agents. However, the effectiveness outcomes used in these studies vary, including achieving IOP thresholds, IOP-controlled days, percent reduction in IOP and QALYs. Methods used to determine costs also vary substantially between studies. Future evaluations of the burden of glaucoma need to consider the issues of comparability between, and generalisability of, study results. Differences in methods have created barriers to understanding the cost of glaucoma and comparing costs or cost effectiveness between studies. Furthermore, future studies should also consider direct costs of glaucoma generally not covered by health insurance as well as indirect costs of glaucoma. As new screening technologies for early detection of individuals at elevated risk of glaucoma are now in use, more complete estimates of the cost of glaucoma are critical for issues of resource allocation and health policy. PMID- 17402804 TI - Pharmacoeconomics of available treatment options for metastatic prostate cancer. AB - The resources devoted to managing metastatic prostate cancer are enormous, yet little attention has been given to directly measuring the economic consequences of treatment alternatives. The purpose of this article was to evaluate the pharmacoeconomics of available treatments for metastatic prostate cancer, including hormone-sensitive disease, androgen-independent prostate cancer and locally advanced/progressive disease. We identified 58 articles addressing economic issues related to metastatic prostate cancer. Treatment alternatives with considerably different costs are available in many areas of disease management, most notably, medical androgen deprivation therapy (ADT) versus surgical castration; combined androgen blockage (CAB) versus monotherapy for initial treatment of hormone-sensitive disease; as well as bisphosphonates and bone-targeted radioisotopes for palliation. The few available pharmacoeconomic studies indicate that the additional costs are not supported by clear and compelling evidence of differences in survival or quality-of-life (QOL) outcomes. Our review revealed that authors often use considerably different assumptions about efficacy and survival outcomes in their analyses, which may be due to the inconsistency of available clinical evidence. Although there have been many clinical trials comparing various therapies, we identified only three trials that included economic assessments. Thus, few sources of economic data are available and most pharmacoeconomic studies rely on information mined from indirect sources. We note that, while there has been considerable enthusiasm about the role of docetaxel regimens in the past 2 years, no study has yet examined the costs of these therapies. Survival remains poor for metastatic disease, thus QOL is the primary consideration for many therapies. However, QOL for treatment of metastatic disease is poorly measured and, in most analyses, the impact of therapy on QOL was inferred based on speculation by the authors. Given the large cost burdens of these treatments, it is essential that we more fully understand the true QOL gains potentially offered by more expensive therapies. The economic studies of advanced prostate cancer highlight several aspects of clinical care that are filled with considerable uncertainty and remain guided by forces other than optimal resource allocation. It is essential that we address the weaknesses in our understanding of the economic consequences of therapies for prostate cancer, and find ways to include economic information into the process of determining optimal therapy. PMID- 17402805 TI - Cost effectiveness of once-daily oral chelation therapy with deferasirox versus infusional deferoxamine in transfusion-dependent thalassaemia patients: US healthcare system perspective. AB - BACKGROUND: Deferasirox is a recently approved once-daily oral iron chelator that has been shown to reduce liver iron concentrations and serum ferritin levels to a similar extent as infusional deferoxamine. OBJECTIVE: To determine the cost effectiveness of deferasirox versus deferoxamine in patients with beta thalassaemia major from a US healthcare system perspective. METHODS: A Markov model was used to estimate the total additional lifetime costs and QALYs gained with deferasirox versus deferoxamine in patients with beta-thalassaemia major and chronic iron overload from blood transfusions. Patients were assumed to be 3 years of age at initiation of chelation therapy and to receive prescribed dosages of deferasirox and deferoxamine that have been shown to be similarly effective in such patients. Compliance with chelation therapy and probabilities of iron overload-related cardiac disease and death by degree of compliance were estimated using data from published studies. Costs ($US, year 2006 values) of deferoxamine administration and iron overload-related cardiac disease were based on analyses of health insurance claims of transfusion-dependent thalassaemia patients. Utilities were based on a study of patient preferences for oral versus infusional chelation therapy, as well as published literature. Probabilistic and deterministic sensitivity analyses were employed to examine the robustness of the results to key assumptions. RESULTS: Deferasirox resulted in a gain of 4.5 QALYs per patient at an additional expected lifetime cost of $US126,018 per patient; the cost per QALY gained was $US28,255. The cost effectiveness of deferasirox versus deferoxamine was sensitive to the estimated costs of deferoxamine administration and the quality-of-life benefit associated with oral versus infusional therapy. Cost effectiveness was also relatively sensitive to the equivalent daily dose of deferasirox, and the unit costs of deferasirox and deferoxamine, and was more favourable in younger patients. CONCLUSION: Results of this analysis of the cost effectiveness of oral deferasirox versus infusional deferoxamine suggest that deferasirox is a cost effective iron chelator from a US healthcare perspective. PMID- 17402806 TI - Therapeutic use of granulocyte colony-stimulating factors for established febrile neutropenia: effect on costs from a hospital perspective. AB - BACKGROUND: The prophylactic use of granulocyte colony-stimulating factors (G CSFs) reduces the severity and duration of neutropenia and reduces the incidence of febrile neutropenia after cancer chemotherapy. However, the use of G-CSFs, particularly filgrastim, to treat established neutropenia remains controversial. A recent meta-analysis of randomised controlled trials (RCTs) evaluating G-CSF treatment for established febrile neutropenia demonstrated a reduction in prolonged hospitalisations. Because more than one-third of patients in the analysis were hospitalised for at least 10 days, this finding has broad pharmacoeconomic and clinical significance. This analysis presents the potential cost implications of G-CSF treatment for established neutropenia among hospitalised patients. METHODS: Direct medical costs ($US, year 2003 values) related to hospitalisation for established neutropenia were modelled using a hospital perspective and according to two treatment options: (i) no use of G-CSF during the neutropenic episode (control); and (ii) addition of daily G-CSF until neutrophil recovery. Within each option, we modelled the probability of a long stay (>or=10 days) and patient survival. The model used three data sets: discharge data from a consortium of academic medical institutions, drug cost data (filgrastim) from Federal payers, and estimates of G-CSF efficacy derived from a meta-analysis of RCTs of treatment in patients with established febrile neutropenia. The lowest expected total cost was predicted for both treatment options; sensitivity analyses and Monte Carlo simulations were used to evaluate the robustness of the model. RESULTS: The G-CSF arm produced the lowest expected cost, and predicted net estimated savings of $US1046 per neutropenic episode compared with the control strategy. G-CSF was less expensive than the control for most reasonable estimates of cost per day and all lengths of stay (LOS) >or=10 days. G-CSF was the least costly strategy for 73.5% of 10,000 Monte Carlo iterations, while the no-G-CSF control strategy predicted savings in 26.5% of iterations. CONCLUSIONS: This pharmacoeconomic model suggests that therapeutic use of G-CSF should be considered for patients with established neutropenia in order to reduce overall hospital cost. G-CSF treatment may offer substantial potential savings for hospitalised patients with established neutropenia over a wide range of model assumptions. Therapeutic G-CSF use among patients hospitalised for established neutropenia may complement the recommended prophylactic use of these agents for the prevention of neutropenic episodes. PMID- 17402808 TI - Causes and consequences of chronic kidney disease: implications for managed health care. AB - BACKGROUND: Chronic kidney disease (CKD) is a major public health problem that often goes unrecognized until late-stage disease. In the United States, nearly 20 million people have CKD, and this number is likely to grow as the population ages and the prevalence of diabetes and hypertension rises. With $28 billion currently spent on end-stage renal disease care in the United States and with the rapidly expanding dialysis population, it is essential to actively address this epidemic, both by reducing the number of patients who reach kidney failure and by decreasing morbidity and mortality among those with early-stage CKD. OBJECTIVE: To review the burden of CKD and its comorbidities on patients, physicians, and payers and discuss the potential benefits to individual patients and society of identifying and treating earlier stages of CKD. CONCLUSION: Major steps in dealing with the CKD epidemic are (1) identifying individuals at risk for and with earlier stages of CKD, (2) initiating therapies to slow progression of kidney disease, and (3) treating comorbid conditions associated with CKD, including cardiovascular disease and anemia. PMID- 17402809 TI - Methods for integrating moderation and mediation: a general analytical framework using moderated path analysis. AB - Studies that combine moderation and mediation are prevalent in basic and applied psychology research. Typically, these studies are framed in terms of moderated mediation or mediated moderation, both of which involve similar analytical approaches. Unfortunately, these approaches have important shortcomings that conceal the nature of the moderated and the mediated effects under investigation. This article presents a general analytical framework for combining moderation and mediation that integrates moderated regression analysis and path analysis. This framework clarifies how moderator variables influence the paths that constitute the direct, indirect, and total effects of mediated models. The authors empirically illustrate this framework and give step-by-step instructions for estimation and interpretation. They summarize the advantages of their framework over current approaches, explain how it subsumes moderated mediation and mediated moderation, and describe how it can accommodate additional moderator and mediator variables, curvilinear relationships, and structural equation models with latent variables. PMID- 17402810 TI - Bias in cross-sectional analyses of longitudinal mediation. AB - Most empirical tests of mediation utilize cross-sectional data despite the fact that mediation consists of causal processes that unfold over time. The authors considered the possibility that longitudinal mediation might occur under either of two different models of change: (a) an autoregressive model or (b) a random effects model. For both models, the authors demonstrated that cross-sectional approaches to mediation typically generate substantially biased estimates of longitudinal parameters even under the ideal conditions when mediation is complete. In longitudinal models where variable M completely mediates the effect of X on Y, cross-sectional estimates of the direct effect of X on Y, the indirect effect of X on Y through M, and the proportion of the total effect mediated by M are often highly misleading. PMID- 17402811 TI - Predicting group-level outcome variables from variables measured at the individual level: a latent variable multilevel model. AB - In multilevel modeling, one often distinguishes between macro-micro and micro macro situations. In a macro-micro multilevel situation, a dependent variable measured at the lower level is predicted or explained by variables measured at that lower or a higher level. In a micro-macro multilevel situation, a dependent variable defined at the higher group level is predicted or explained on the basis of independent variables measured at the lower individual level. Up until now, multilevel methodology has mainly focused on macro-micro multilevel situations. In this article, a latent variable model is proposed for analyzing data from micro-macro situations. It is shown that regression analyses carried out at the aggregated level result in biased parameter estimates. A method that uses the best linear unbiased predictors of the group means is shown to yield unbiased estimates of the parameters. PMID- 17402812 TI - Item factor analysis: current approaches and future directions. AB - The rationale underlying factor analysis applies to continuous and categorical variables alike; however, the models and estimation methods for continuous (i.e., interval or ratio scale) data are not appropriate for item-level data that are categorical in nature. The authors provide a targeted review and synthesis of the item factor analysis (IFA) estimation literature for ordered-categorical data (e.g., Likert-type response scales) with specific attention paid to the problems of estimating models with many items and many factors. Popular IFA models and estimation methods found in the structural equation modeling and item response theory literatures are presented. Following this presentation, recent developments in the estimation of IFA parameters (e.g., Markov chain Monte Carlo) are discussed. The authors conclude with considerations for future research on IFA, simulated examples, and advice for applied researchers. PMID- 17402813 TI - Mixture distribution latent state-trait analysis: basic ideas and applications. AB - Extensions of latent state-trait models for continuous observed variables to mixture latent state-trait models with and without covariates of change are presented that can separate individuals differing in their occasion-specific variability. An empirical application to the repeated measurement of mood states (N=501) revealed that a model with 2 latent classes fits the data well. The larger class (76%) consists of individuals whose mood is highly variable, whose general well-being is comparatively lower, and whose mood variability is influenced by daily hassles and uplifts. The smaller class (24%) represents individuals who are rather stable and happier and whose mood is influenced only by daily uplifts but not by daily hassles. A simulation study on the model without covariates with 5 sets of sample sizes and 5 sets of number of occasions revealed that the appropriateness of the parameter estimates of this model depends on number of observations (the higher the better) and number of occasions (the higher the better). Another simulation study estimated Type I and II errors of the Lo-Mendell-Rubin test. PMID- 17402814 TI - On the consistency of individual classification using short scales. AB - Short tests containing at most 15 items are used in clinical and health psychology, medicine, and psychiatry for making decisions about patients. Because short tests have large measurement error, the authors ask whether they are reliable enough for classifying patients into a treatment and a nontreatment group. For a given certainty level, proportions of correct classifications were computed for varying test length, cut-scores, item scoring, and choices of item parameters. Short tests were found to classify at most 50% of a group consistently. Results were much better for tests containing 20 or 40 items. Small differences were found between dichotomous and polytomous (5 ordered scores) items. It is recommended that short tests for high-stakes decision making be used in combination with other information so as to increase reliability and classification consistency. PMID- 17402815 TI - Vascular health and longitudinal changes in brain and cognition in middle-aged and older adults. AB - The impact of vascular health on the relations between structural brain changes and cognition was assessed in a longitudinal study of 46 adults, 23 of whom remained healthy for 5 years and 23 of whom had hypertension at baseline or acquired vascular problems during follow-up. At both measurement occasions, the volume of white matter hyperintensities (WMH) and regional brain volumes correlated with age. In 5 years, WMH volume more than doubled in the vascular risk group but did not increase in healthy participants. The frontal lobes had the highest WMH load at baseline and follow-up; the parietal WMH showed the greatest rate of expansion. In the vascular risk group, systolic blood pressure at follow-up correlated with posterior WMH volume. The fastest cortical shrinkage was observed in the prefrontal cortex and the hippocampus. Fluid intelligence correlated with WMH burden and declined along with faster WMH progression. In the vascular risk group, WMH progression and shrinkage of the fusiform cortex correlated with decline in working memory. Thus, poor vascular health contributes to age-related declines in brain and cognition, and some of the age-related declines may be limited to persons with elevated vascular risk. PMID- 17402816 TI - Longitudinal change in cognitive performance among individuals with mild cognitive impairment. AB - The authors used mixed-effects growth models to examine longitudinal change in neuropsychological performance over a 4-year period among 197 individuals who were either normal or had mild cognitive impairment (MCI) at baseline. At follow up, the participants were divided into 4 groups: (a) controls: participants who were normal at both baseline and follow-up (n = 33), (b) stables: participants with MCI whose Clinical Dementia Rating-Sum of Boxes (CDR-SB) score did not differ between the first and last evaluations (n = 22), (c) decliners: participants with MCI whose CDR-SB score declined between the first and last evaluations (n = 95), and (d) converters: participants who received a clinical diagnosis of Alzheimer's disease during the follow-up period (n = 47). Only the Episodic Memory factor showed a significantly greater rate of decline over the follow-up period among the converters. Two other factors were significantly lower in converters at baseline in comparison with other groups (the executive function factor and the general knowledge factor), but the rate of decline over time did not differ. Individuals with an APOE epsilon4 allele scored lower on the episodic memory and executive function factors at baseline. PMID- 17402817 TI - Spatial attention and response control in healthy younger and older adults and individuals with Alzheimer's disease: evidence for disproportionate selection impairments in the Simon task. AB - The authors examined the degree to which aging and Alzheimer's disease (AD) influence the ability to control attention when conflict is presented in terms of incongruent mapping between a stimulus and the appropriate response. In a variant of the Simon task, healthy older adults and older adults with mild or very mild AD showed disproportionately larger reaction time (RT) costs when the stimulus and response were in conflict relative to RT costs of healthy younger adults. Analyses of RT distributions provide support for a 2-process model of the Simon effect in which there is a short-lived transient effect of the irrelevant dimension in younger adults and a more sustained influence across the RT distribution in older adults. An analysis of error rates showed that the older adults with mild and very mild AD made more errors on incongruent trials, suggesting that AD leads to increased likelihood of selecting the prepotent pathway. The findings are discussed in terms of the special nature of the response requirements of the Simon task to better illuminate the attentional decrements in both healthy aging and early stage AD. PMID- 17402818 TI - Implicit category learning performance predicts rate of cognitive decline in nondemented patients with Parkinson's disease. AB - Nondemented patients with Parkinson's disease (PD) are impaired in learning to categorize simple perceptual stimuli when category membership is defined by a nonlinear relationship between stimulus dimensions but not when the relationship is linear (J. V. Filoteo, W. T. Maddox, D. P. Salmon, & D. D. Song, 2005). In the present study, the authors examined whether performance in either of these 2 category learning conditions was predictive of global cognitive decline following a mean of 1.6 years since the time patients were 1st seen. Results indicated that final block accuracy in the nonlinear condition, but not the linear condition, predicted global cognitive decline. Performance on the Wisconsin Card Sorting Test (WCST) did not significantly predict global cognitive decline, although there was a trend for this to be the case. In addition, the association between nonlinear category learning and global cognitive decline was not impacted by patients' performance on the WCST. Results suggest that nonlinear category learning predicts cognitive decline in nondemented patients with PD and that nonlinear category learning and WCST performances may provide independent measures of integrity of the posterior and anterior caudate, respectively. PMID- 17402819 TI - Categorization of novel animals by patients with Alzheimer's disease and corticobasal degeneration. AB - We taught a novel animal category by rule-based and similarity-based processes to participants with Alzheimer's disease (AD), corticobasal degeneration (CBD), and healthy age-matched participants. Healthy participants successfully categorized by either process. AD patients' rule-based categorization was impaired, while their similarity-based categorization resembled that of healthy participants. Correlations of AD patients' performance with measures of executive functioning suggested a deficit in the cognitive resources necessary for engaging rule-based categorization. The contribution of limited executive resources to categorization difficulty in AD was further demonstrated in a second experiment in which features determining category membership were of lower salience. CBD patients were relatively impaired at similarity-based processing, suggesting that qualitatively distinct categorization processes can be selectively compromised in patients with focal neurodegenerative diseases. Moreover, AD patients' impaired categorization correlated with performance on a measure of semantic memory, implicating this categorization deficit in AD patients' semantic memory difficulty. PMID- 17402820 TI - Gender-related dissociations of categorical fluency in normal subjects and in subjects with Alzheimer's disease. AB - Semantic fluency tasks, with the categories of birds and furniture as stimuli, were administered to normal subjects and patients with Alzheimer's disease (AD). Normal subjects showed a gender-related double dissociation consistent with the literature data because men were more fluent with the birds category and women with the furniture category. Also, patients with AD showed a Gender x Category interaction, but the double dissociation between birds and furniture was not present because of a prevalent impairment of the living category birds that was irrespective of gender. This pattern of impairment in patients with AD was independent from the disease stage. The authors conclude that (a) gender-related categorical effects cannot be considered as inborn, sex-related cognitive differences but as familiarity effects and (b) both lesion-related and familiarity-related factors must be taken into account to explain category specific effects of patients with brain damage and patients with AD. PMID- 17402821 TI - Changes in attention and information-processing speed following severe traumatic brain injury: a meta-analytic review. AB - Deficits in attention are frequently reported following severe traumatic brain injury (TBI). However, methodological differences make it difficult to reconcile inconsistencies in the research findings in order to undertake an evidence-based assessment of attention. The current study therefore undertook a meta-analytic review of research examining attention following severe TBI. A search of the PsycINFO and PubMed databases spanning the years 1980 to 2005 was undertaken with 24 search terms. Detailed inclusion and exclusion criteria were used to screen all articles, leaving 41 studies that were included in the current meta-analysis. Weighted Cohen's d effect sizes, percentage overlap statistics, and confidence intervals were calculated for the different tests of attention. Fail-safe Ns were additionally calculated to address the bias introduced by the tendency to publish significant results. Large and significant deficits were found in specific measures of information-processing speed, attention span, focused/selective attention, sustained attention, and supervisory attentional control following severe TBI. Finally, age, education, and postinjury interval were not significantly related to these deficits in attention. PMID- 17402822 TI - Feeling of knowing in episodic memory following moderate to severe closed-head injury. AB - The ability to accurately monitor one's memory is a metacognitive process that is important in everyday life. The authors examined episodic memory feeling-of knowing (FOK) ratings in 21 moderate to severe closed-head injury (CHI) participants (more than 1 year postinjury) and 21 controls. Participants studied 36 critical cue-target word pairs. Following a brief delay, they were asked to recall the target that corresponded to a given cue. Confidence ratings were made for recalled words, and FOK judgments were made for nonrecalled words in terms of the likelihood of recognizing the target word on a subsequent recognition test. CHI participants demonstrated less accurate recall but accurate ability to judge their recall performance (retrospective memory monitoring). They also demonstrated intact FOK judgments when providing binary judgments but demonstrated difficulties making finer discriminations on an ordinal scale (prospective memory monitoring). These findings suggest that memory monitoring is not a unitary construct. It is proposed that CHI participants may display intact memory monitoring when predictions are based on familiarity assessment but not when continued probing for additional episodic information is required. PMID- 17402823 TI - Reading and the corpus callosum: an MRI family study of volume and area. AB - This study examines the corpus callosum in 68 readers nested in 24 families. Callosa were measured and controlled for whole brain volume, intelligence, and gender. The relation of corpus callosum size to the within-family variance of oral reading was investigated with various measurements: volume, midsagittal area, and anterior-to-posterior one-fifth area segmentations. Because this is the first known publication of MRI calculations of corpus callosum volume, some basic questions about bilateral symmetry and the efficacy of area versus volumetric measurements were explored. Results suggest that better readers within families have larger midsagittal areas at the midbody. Although reliably measured, volume did not contribute to oral reading but was highly correlated with area. Bilateral volumes of the corpus callosum were symmetric. PMID- 17402824 TI - Temporal dynamics of visuo-tactile extinction within and between hemispaces. AB - The ability to detect left-sided stimuli (visual or tactile) was studied in visual and tactile extinction (RBD) patients and in healthy (C) subjects. Stimuli were single or double; double stimuli were always cross-modal and could be released across the 2 hemispaces or within the same hemispace. Moreover, the stimuli could be either simultaneous or separated by 1 of 3 different asynchronies (105, 505, 905 ms). C subjects were perfect in all conditions. RBD patients omitted the contralesional tactile stimulus in bilateral trials (across space, classical extinction). They also omitted the tactile stimulus in unilateral left trials (within hemispace). Both effects showed the same temporal modulation with lower extinction rates at longer stimulus onset asynchronies. Results suggest that attentional processing of a visual stimulus inhibits processing of a tactile one, even if both are delivered in the same contralesional hemispace. PMID- 17402825 TI - Stroop interference and attention-deficit/hyperactivity disorder: a review and meta-analysis. AB - Previous reviews and meta-analyses that addressed abnormal Stroop interference in attention-deficit/hyperactivity disorder (ADHD) yielded mixed results. The authors of the present study argue that the inconsistencies may reflect the problematic nature of 2 frequently used methods to quantify Stroop interference the difference score and Golden's method (C. J. Golden, 1978). Golden's method correction for base-word reading is inadequate, and the difference score is sensitive to the nature of the outcome variable. The latter can be remedied with a ratio score. Contrasting previous meta-analyses, this meta-analysis covers all age groups and all Stroop test variants, and it excludes studies using the Golden quantification method. Mean effect sizes for interference in ADHD as quantified by difference scores relative to control scores were 0.24 across all studies but 1.11 for time-per-item studies; outcome variable was a significant moderator variable, reflecting the sensitivity of the difference score to this variable. Consistency analysis of ratio scores across 19 studies reveals more interference for the ADHD groups relative to the control groups. It is concluded that interference control is consistently compromised in individuals with ADHD. PMID- 17402826 TI - Neuropsychological functioning of girls with attention-deficit/hyperactivity disorder followed prospectively into adolescence: evidence for continuing deficits? AB - The current study prospectively followed girls with attention deficit/hyperactivity disorder (ADHD), along with a matched comparison sample, 5 years after childhood neuropsychological assessments. Follow-up neuropsychological measures emphasized attentional skills, executive functions, and language abilities. Paralleling childhood findings, the childhood-diagnosed ADHD group displayed moderate to large deficits in executive/attentional performance as well as in rapid naming relative to the comparison group at follow up (M-sub(age) 14.2 years). ADHD-inattentive versus ADHD-combined contrasts were nonsignificant and of negligible effect size, even when a refined, sluggish cognitive tempo subgroup of the inattentive type was examined. Although ADHD versus comparison group differences largely withstood statistical control of baseline demographics and comorbidities, control of childhood IQ reduced executive function differences to nonsignificance. Yet when the subset of girls meeting diagnostic criteria for ADHD in adolescence was compared with the remainder of the participants, neuropsychological deficits emerged even with full statistical control. Overall, childhood ADHD in girls portends neuropsychological and executive deficits that persist for at least 5 years. PMID- 17402828 TI - Antimicrobial stewardship: the aspirin of the 21st century. PMID- 17402829 TI - Surrogate markers to assess new therapies for drug-resistant tuberculosis. PMID- 17402830 TI - Vietnam aims to alleviate HIV prevalence. PMID- 17402832 TI - New frontiers in tuberculosis research. PMID- 17402833 TI - Daptomycin: a rapidly bactericidal lipopeptide for the treatment of Gram-positive infections. AB - Antimicrobial resistance among Gram-positive organisms continues to increase and has reached epidemic proportions in a number of countries and within medical centers worldwide. Daptomycin is a new lipopeptide antibiotic with rapid bactericidal activity against Staphylococcus aureus. It is also active against coagulase-negative staphylococci, enterococci and streptococci. It exerts its effect through cell membrane disruption that results in dissipation of the membrane potential. Daptomycin exhibits a prolonged postantibiotic effect and is well tolerated. In Phase III clinical trials, daptomycin was found to be similar in efficacy to standard therapy in complicated skin and skin structure infections. More recently, it was approved for the treatment of S. aureus bacteremia and right-sided endocarditis. Daptomycin is not indicated for pulmonary infections. Preliminary data suggest that daptomycin may be effective in urinary tract, bone and joint infections. However, randomized clinical trials are needed to confirm these findings. Daptomycin is an effective antimicrobial agent for the treatment of various serious Gram-positive infections, especially those caused by methicillin-resistant S. aureus. PMID- 17402834 TI - Faropenem: review of a new oral penem. AB - Faropenem medoxomil is a new orally administered penem antibiotic. Its chiral tetrahydrofuran substituent at position C2 is responsible for its improved chemical stability and reduced CNS effects, compared with imipenem. Faropenem demonstrates broad-spectrum in vitro antimicrobial activity against many Gram positive and -negative aerobes and anaerobes, and is resistant to hydrolysis by nearly all beta-lactamases, including extended-spectrum beta-lactamases and AmpC beta-lactamases. However, faropenem is not active against methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, Pseudomonas aeruginosa or Stenotrophomonas maltophilia. Prospective, multicenter, randomized, double-blind, comparative (not vs placebo) clinical trials of acute bacterial sinusitis (ABS), acute exacerbations of chronic bronchitis (AECB), community acquired pneumonia (CAP) and uncomplicated skin and skin structure infections (uSSSIs) have demonstrated that faropenem medoxomil has equivalent efficacy and safety compared with cefuroxime, clarithromycin, azithromycin, amoxicillin, cefpodoxime and amoxicillin-clavulanate. The evidence supports faropenem medoxomil as a promising new oral beta-lactam with proven efficacy and safety for the treatment of a variety of community-acquired infections. However, the US FDA recently rejected faropenem for all four indications stating that the clinical trials in ABS and AECB should have been performed versus a placebo. In the CAP studies, the FDA stated that they could not be certain of the validity of the study population actually having the disease and for uSSSI, the FDA stated that only a single trial was not adequate evidence of efficacy for this indication. PMID- 17402835 TI - Artesunate, artemether or quinine in severe Plasmodium falciparum malaria? AB - Quinine and the artemisinin-derivative drugs artesunate and artemether are effective treatments for severe falciparum malaria. Trials comparing artemether with quinine have not demonstrated convincing evidence of a mortality advantage for artemether. The South East Asian Quinine Artesunate Malaria Trial (SEAQUAMAT), a multicenter, randomized, open-label trial in 1461 adults with severe malaria in Asia compared artesunate with quinine. Mortality was 15% in the artesunate group and 22% in the quinine group, a reduction of 34.7% (95% confidence interval: 18.5-47.6%) in the artesunate group, with almost all the benefit reported in those with high parasite counts. Artesunate should constitute first-line treatment for severe malaria in Asia. These results can probably be generalized to the treatment of severe malaria in adults from all areas, especially in those with hyperparasitemia. However, it is unclear whether these results can be generalized to children in Africa, who constitute the majority of those who die from severe malaria worldwide. PMID- 17402836 TI - A new era in HIV vaccine development. AB - Since the identification of HIV in 1984, the search for a safe and effective vaccine has been relentless. While investigator-initiated research has provided substantial information regarding HIV disease and pathogenesis, and over two dozen drugs are licensed in the USA to treat HIV, the global epidemic continues unabated. Early in HIV vaccine research, the pharmaceutical industry took the initiative to produce products for clinical testing. As the likelihood of a quick success decreased, private investment waned. The public sector responded with novel mechanisms to engage industry while continuing to support academic investigators. HIV vaccine research continues to rely on the creativity of individual investigators, as well as collaborations that vary in size and complexity and offer opportunities for the efficient use of resources and accelerated progress. PMID- 17402837 TI - Current non-AIDS antiviral chemotherapy. AB - The evolution of antiviral therapy began with developments in the management of influenza and herpes simplex keratitis in the 1960s and early 1970s. However, the field exploded with the successful treatment of herpes simplex encephalitis, herpes zoster and genital herpes simplex virus infections, all occurring in the late 1970s and early 1980s. These advances have contributed to the development of therapies for HIV that have transformed the lives of infected patients in recent years. The clinical fruit of all of these research advances has been an armamentarium of drugs that can be used to successfully treat a variety of viral illnesses. In addition to HIV/AIDS, current antiviral therapy focuses primarily on herpesviruses, hepatitis viruses and influenza. Notably, considerable progress remains to be made in these areas. Moreover, a variety of additional viral diseases currently require the development of specific therapies. PMID- 17402838 TI - Interactions and management issues in HSV and HIV coinfection. AB - Significant synergistic interactions have been observed between HIV and herpes simplex virus (HSV). HIV-induced immune compromise can cause frequent and persistent HSV disease, while poorly controlled HSV replication may influence HIV pathogenicity and transmission. HSV-2 seroprevalence is high in HIV-infected cohorts worldwide, with rates of over 80% for HSV-1 and ranging from 33% to more than 80% for HSV-2. As seen in HIV-negative individuals, HSV-2 coinfection is associated with female gender, older age and black ethnicity. HSV infection is commonly under-diagnosed in HIV-infected individuals, although the use of PCR for HSV detection in mucocutaneous swabs and HSV type-specific serology can improve the diagnostic yield. In HIV-1-infected patients with frequent clinical episodes of HSV reactivation, suppressive antiviral therapy may prove beneficial in controlling HSV disease while also reducing HSV-mediated promotion of HIV replication. Antiretroviral therapy leads to a gradual recovery of HSV-specific T cell responses and a reduction in HSV-related morbidity, indicating that successful management of coinfection should target both HIV and HSV replication. The aim of this review is to address the more speculative issues surrounding the management of HSV/HIV coinfection and to summarize the data that inform them. PMID- 17402839 TI - Identification of new human coronaviruses. AB - To date, there are still a variety of human infections with unknown etiology. Identification of previously unrecognized viral agents in patient samples is of great medical interest but remains a major technical challenge. Acute respiratory tract infections are responsible for considerable morbidity and mortality in humans and animals. A variety of viruses, bacteria and fungi are associated with respiratory tract illness. Most of the respiratory viruses belong to the Paramyxoviridae, Orthomyxoviridae, Picornaviridae, Adenoviridae and Coronaviridae families. No pathogens can be detected in a relatively large proportion of patients with respiratory disease, partially owing to limitations of current diagnostic assays but also since some infections are caused by as yet unknown pathogens. This review will focus on human coronaviruses. In the mid 1960s, two human coronaviruses were identified that cause the common cold: human coronaviruses (HCoV)-229E and HCoV-OC43. The recent outbreak of severe acute respiratory syndrome-CoV and subsequent identification of two additional human coronaviruses (HCoV-NL63 and HCoV-HKU1) has drawn attention to this virus family. This review summarizes the knowledge of current methodologies for identifying novel human coronavirus species. Furthermore, information on the discovery of known human coronaviruses will be presented. PMID- 17402840 TI - Pharmacodynamics of antibiotics to treat multidrug-resistant Gram-positive hospital infections. AB - Hospital infection due to multidrug-resistant Gram-positive bacteria may often represent a life-threatening challenge; thus, to appropriately combat them, clinicians should be confident and systematically apply several important pharmacodynamic concepts. The concept of 'correct antibiotic treatment' should include, in addition to an appropriate choice in terms of antimicrobial susceptibility, an appropriate dosage and administration schedule consistent with the pharmacodynamic principles. In the next few years, it is expected that some of the anti-Gram-positive antimicrobial agents that are currently under investigation will be added to the therapeutic armamentarium. However, optimization of the usage of old drugs still remains a clinical priority and a scientific challenge, whose dignity may be considered of similar importance to that of the assessment of the possible therapeutic role of the new compounds. PMID- 17402842 TI - Candida urinary tract infections: treatment options. AB - Candiduria is a nonspecific finding that occurs with contamination of a urine sample, colonization of an indwelling catheter and/or the bladder, symptomatic cystitis and invasive upper tract infection. Most patients are colonized and do not require antifungal therapy. Removing predisposing factors, such as indwelling catheters and antibiotics, will clear candiduria in almost 50% of asymptomatic patients. For patients with symptomatic Candida urinary tract infections, a variety of treatment options are available. Fluconazole is the antifungal agent of choice, achieving high urine concentrations with the oral formulation. Rarely, amphotericin B or flucytosine are used. Newer azole agents and echinocandins are not recommended for the treatment of urinary tract infections since they fail to achieve adequate urine concentrations. PMID- 17402841 TI - Novel approaches to bacterial infection therapy by interfering with bacteria-to bacteria signaling. AB - The growing challenge of antimicrobial resistance and the paucity of novel antibiotics underscore the importance of developing novel therapeutics. Bacterial cell-to-cell signaling constitutes a novel drug target. Quorum sensing (QS) is a cell-to-cell signaling mechanism that refers to the ability of bacteria to respond to chemical hormone-like molecules called autoinducers. QS is responsible for controlling a plethora of virulence genes in several bacterial pathogens. Antagonists to autoinducers will intercept bacterial intercellular communication, hindering their ability to act in a coordinated manner to express virulence traits. Moreover, since QS is not involved directly in essential processes, such as bacterial growth, one can reason that inhibition of QS will not yield a selective pressure for the development of resistance. PMID- 17402843 TI - Prevention and treatment of congenital toxoplasmosis. AB - Infection with Toxoplasma gondii is transmitted to man by infected meat or meat products and by contact with soil or surface water. In theory, prevention by hygienic measures is possible, but this has never been proved to work in practice. Therefore, pre- and postnatal screening has been implemented in several countries aiming at early diagnosis. However, data on the effect of treatment are limited and no randomized, controlled trials have been performed. The risk of T. gondii infection in Europe is declining and studies using historical controls from earlier decades cannot be used for decision making. The screening of pregnant women or neonates makes the assumption that any children diagnosed can be offered an effective treatment. There is an urgent need to test new drugs and demonstrate, using randomized, controlled trials, that the currently used drugs are effective. PMID- 17402844 TI - Prevention and management of cytomegalovirus infection in solid-organ transplantation. AB - Cytomegalovirus (CMV) is the most common opportunistic viral infection to occur following solid-organ transplantation. This review will discuss the current strategies of management of CMV in solid-organ transplantation and their challenges. There are two principal approaches for preventing CMV disease in recipients of solid-organ transplants: prophylactic and pre-emptive. Ganciclovir is the most studied and used antiviral for both treatment and prevention, and is the first-line treatment for CMV infection and CMV disease in transplant recipients. There is no consensus regarding the most appropriate prevention method and the approach to CMV disease prevention differs among transplantation centers owing to the paucity of data comparing the two strategies head-to-head. Currently, the recommended treatment for CMV disease is intravenous ganciclovir. PMID- 17402845 TI - Prevention and treatment of aortic graft infection. AB - Prosthetic vascular graft infection remains one of the most challenging surgical problems for vascular surgeons. This condition is classically associated with high morbidity and mortality rates. Accurate diagnosis of a vascular graft infection can typically be made based on a thorough history and physical examination; although, infrequently, an extensive radiological evaluation is necessary to establish the clinical finding. Complete graft excision and extra anatomic bypass grafting remains a commonly accepted surgical treatment strategy. Recent clinical data have supported other treatment modalities, including the use of in situ antibiotic-impregnated graft replacement, in situ allograft replacement and in situ autologous graft replacement. This article will review the pathobiology of aortic graft infection, as well as methods to prevent a prosthetic graft infection. Furthermore, various surgical treatment modalities of aortic graft infection will be discussed. PMID- 17402846 TI - Induction of DC-STAMP by alternative activation and downstream signaling mechanisms. AB - DC-STAMP is essential for fusion of osteoclasts and foreign body giant cells; however, it is not known whether dc-stamp expression in these two cell types is differentially regulated. Here, we show that dc-stamp expression and cell-cell fusion are regulated in a cell type-specific manner. INTRODUCTION: The transcription factors c-Fos and NFATc1 cooperate to regulate osteoclast differentiation, whereas PU.1 and NF-kappaB are activated in macrophages and osteoclasts or in both cell types. Thus, we asked what role c-Fos, NFATc1, PU.1, and NF-kappaB played in regulating dendritic cell-specific transmembrane protein (dc-stamp) expression and fusion of osteoclasts and macrophage giant cells. MATERIALS AND METHODS: Transcriptional activation by c-Fos and NFATc1 was examined by dc-stamp promoter analysis. Multinuclear cell formation was analyzed in cells from c-Fos-deficient mice or in wildtype cells treated with the NFAT inhibitor FK506. The role of DC-STAMP in cell fusion was examined in vitro in a macrophage giant cell formation assay using DC-STAMP-deficient cells. Recruitment of c-Fos, NFATc1, PU.1, and NF-kappaB to the dc-stamp promoter in osteoclasts and macrophage giant cells was analyzed by chromatin-immunoprecipitation analysis. RESULTS: Both activator protein-1 (AP-1) and NFAT binding sites in the dc-stamp promoter were needed for dc-stamp expression after RANKL stimulation of osteoclasts. dc-stamp expression was induced in osteoclasts and macrophage giant cells, and cells from DC-STAMP-deficient mice failed to form either multinuclear osteoclasts or macrophage giant cells. In contrast, c-Fos is indispensable for dc stamp expression and cell-cell fusion under conditions favoring in vitro and in vivo induction of osteoclasts but not macrophage giant cells. Consistently, an NFAT inhibitor suppressed multinuclear osteoclast formation but not macrophage giant cell formation. In addition, PU.1 and NF-kappaB binding sites were detected in the dc-stamp promoter, and both PU.1 and NF-kappaB were recruited to the dc stamp promoter after granulocyte-macrophage colony stimulating factor (GM-CSF) + interleukin (IL)-4 stimulation. CONCLUSIONS: dc-stamp expression is regulated differently in osteoclasts and macrophage giant cells. c-Fos and NFATc1, both of which are essential for osteoclast differentiation, are needed for dc-stamp expression and cell-cell fusion in osteoclasts, but both factors are dispensable for giant cell formation by macrophages. Because PU.1 and NF-kappaB are recruited to the dc-stamp promoter after stimulation with GM-CSF + IL-4, dc-stamp transcription is regulated in a cell type-specific manner. PMID- 17402847 TI - Accretion of bone quantity and quality in the developing mouse skeleton. AB - In this work, we found that bone mineral formation proceeded very rapidly in mice by 1 day of age, where the degree of mineralization, the tissue mineral density, and the mineral crystallinity reached 36%, 51%, and 87% of the adult values, respectively. However, even though significant mineralization had occurred, the elastic modulus of 1-day-old bone was only 14% of its adult value, indicating that the intrinsic stiffening of the bone lags considerably behind the initial mineral formation. INTRODUCTION: To meet the mechanical challenges during early development, the skeleton requires the rapid accretion of bone quality and bone quantity. Here, we describe early bone development in the mouse skeleton and test the hypothesis that specific compositional properties determine the stiffness of the tissue. MATERIALS AND METHODS: Tibias of female BALB mice were harvested at eight time-points (n = 4 each) distributed between 1 and 40 days of age and subjected to morphometric (muCT), chemical (Fourier transform infrared microspectroscopy), and mechanical (nanoindentation) analyses. Tibias of 450-day old mice served as fully mineralized control specimens. RESULTS: Bone growth proceeded very rapidly; at 1 day of age, the degree of mineralization (phosphate/protein ratio), the density of mineralized bone (TMD), and mineral crystallinity had reached 36%, 51%, and 87% of the adult (450 days) values, respectively. Spatially, the variability in mineralization across the mid diaphysis was very high for the early time-points and declined over time. In contrast to the notable changes in mineralization, carbonate substitution into the mineral lattice (carbonate/phosphate ratio) and collagen cross-linking did not show any significant changes over this time period. Even though significant mineralization had occurred, the elastic modulus of 1-day-old bone was only 14% of the adult value and increased to 89% (of its adult value) after 40 days. Between samples of different time-points, significant positive correlations were observed between the elastic modulus and TMD (r(2) = 0.84), phosphate/protein ratio (r(2) = 0.59), and crystallinity (r(2) = 0.23), whereas collagen cross linking showed a small but significant negative correlation (r(2) = 0.15). CONCLUSIONS: These data indicate that specific chemical and morphometric properties modulate bone's stiffness during early growth. The intrinsic stiffening of the bone, however, lags considerably behind the initial mineral formation, emphasizing the importance of bone mineral quality for optimizing matrix integrity. PMID- 17402848 TI - How should we evaluate bone mass in children? PMID- 17402850 TI - Relative risk of mortality after traumatic brain injury: a population-based study of the role of age and injury severity. AB - To test if observed vs. expected mortality differs by age among traumatic brain injury (TBI) cases, a population-based, historical cohort study was conducted in Olmsted County, Minnesota. From all residents with any diagnosis suggestive of TBI 1985-1999, we randomly sampled 7,800 and reviewed their medical records to confirm the event. Confirmed incident cases were categorized by age in years (<16 = pediatric, 16-65 = adult, > 65 elderly) and severity (moderate/severe vs. mild) and followed for vital status through 6/30/2004. We compared observed 6-month and 10-year mortality with expected and tested if the differences varied by age. Of 1,433 confirmed incident cases, 35% were pediatric; 55% were adult; only 9% were elderly; 11.2% of all cases were moderate/severe; the proportions by increasing age group were 11.4%, 8.5%, 26.7%. The proportions who died within 6 months increased with increasing age group, both for moderate/severe (10.3%, 40.3%, 50.0%) and mild cases (0%, 0%, 9.1%); mortality for moderate/severe cases was nearly 40 times that for mild cases, independent of age. Among 6-month survivors, 10-year mortality differed from expected only for adult cases. For all cases, after adjusting for sex, year of TBI, and severity, the difference between observed and expected 10-year mortality was greater for adult cases than for pediatric cases and similar for adult and elderly cases. Elderly individuals account for <10% of TBI cases and >50% of 10-year mortality, yet much of this discrepancy reflects age-associated mortality in general. Findings have implications for (1) reducing the number of excess deaths following TBI and (2) caring for survivors. PMID- 17402851 TI - Global white matter analysis of diffusion tensor images is predictive of injury severity in traumatic brain injury. AB - Conventional clinical neuroimaging is insensitive to axonal injury in traumatic brain injury (TBI). Immunocytochemical staining reveals changes to axonal morphology within hours, suggesting potential for diffusion-weighted magnetic resonance (MR) in early diagnosis and management of TBI. Diffusion tensor imaging (DTI) characterizes the three-dimensional (3D) distribution of water diffusion, which is highly anisotropic in white matter fibers owing to axonal length. Recently, DTI has been used to investigate traumatic axonal injury (TAI), emphasizing regional analysis in more severe TBI. In the current study, we hypothesized that a global white matter (WM) analysis of DTI data would be sensitive to TAI across a spectrum of TBI severity and injury to scan interval. To investigate this, we compared WM-only histograms of a scalar, fractional anisotropy (FA), between 20 heterogeneous TBI patients recruited from Detroit Medical Center, including six mild TBI (GCS 13-15), and 14 healthy age-matched controls. FA histogram parameters were correlated with admission GCS and posttraumatic amnesia (PTA). In all cases, including mild TBI, patients' FA histograms were globally decreased compared with control histograms. The shape of the TBI histograms also differed from controls, being more peaked and skewed. The mean FA, kurtosis and skewness were highly correlated suggesting a common mechanism. FA histogram properties also correlated with injury severity indexed by GCS and PTA, with mean FA being the best predictor and duration of PTA (r = 0.64) being superior to GCS (r = 0.47). Therefore, in this heterogeneous sample, the FA mean accounted for 40% of the variance in PTA. Increased diffusion in the short axis dimension, likely reflecting dysmyelination and swelling of axons, accounted for most of the FA decrease. FA is globally deceased in WM, including mild TBI, possibly reflecting widespread involvement. FA changes appear to be correlated with injury severity suggesting a role in early diagnosis and prognosis of TBI. PMID- 17402852 TI - Calpain-mediated collapsin response mediator protein-1, -2, and -4 proteolysis after neurotoxic and traumatic brain injury. AB - Collapsin response mediator proteins (CRMPs) are important molecules in neurite outgrowth and axonal guidance. Within the CRMP family, CRMP-2 has been implicated in several neurological diseases (Alzheimer's, epilepsy, and ischemia). Here, we investigated the integrity of CRMPs (CRMP-1, -2, -4, -5) after in vitro neurotoxin treatment and in vivo traumatic brain injury (TBI). After maitotoxin (MTX) and NMDA treatment of primary cortical neurons, a dramatic decrease of intact CRMP-1, -2 and -4 proteins were observed, accompanied by the appearance of distinct 55-kDa and 58-kDa breakdown products (BDP) for CRMP-2 and -4, respectively. Inhibition of calpain activation prevented NMDA-induced CRMP-2 proteolysis and redistribution of CRMP-2 from the neurites to the cell body, while attenuating neurite damage and neuronal cell injury. Similarly, CRMP-1, -2, and -4 were also found degraded in rat cortex and hippocampus following controlled cortical impact (CCI), an in vivo model of TBI. The appearance of the 55-kDa CRMP-2 BDP was observed to increase, in a time-dependent manner, between 24 and 48 h in the ipsilateral cortex, and by 48 hours in the hippocampus. The observed 55-kDa CRMP-2 BDP following TBI was reproduced by in vitro incubation of naive brain lysate with activated calpain-2, but not activated caspase-3. Sequence analysis revealed several possible cleavage sites near the C-terminus of CRMP-2. Collectively, this study demonstrated that CRMP-1, -2, and -4 are degraded following both acute traumatic and neurotoxic injury. Furthermore, calpain-2 was identified as the possible proteolytic mediator of CRMP-2 following excitotoxic injury and TBI, which appears to correlate well with neuronal cell injury and neurite damage. It is possible that the calpain-mediated truncation of CRMPs following TBI may be an inhibiting factor for post-injury neurite regeneration. PMID- 17402853 TI - Gender differences in spinal cord injury are not estrogen-dependent. AB - Recent attention has been given to gender differences in neurotrauma, and the anecdotal suggestion is that females have better outcomes than males, suggesting that circulating levels of estrogen (E(2)) may be neuroprotective. In order to address this issue, both young adult male and ovariectomized female rats were subjected to a T10 spinal cord injury (SCI), and E2 levels were maintained at chronic, constant circulating levels. Animals were clinically evaluated for locomotor changes using the Basso-Beattie-Bresnahan (BBB) scoring system. Morphologic differences were evaluated with unbiased stereology. Data analysis failed to reveal any significant benefit for the E2 therapy in either males or females. We did find a non-estrogen-dependent difference between male and female rats in length of injury, and percent of spared tissue, with female outcomes more favorable. These results suggest that E(2) does not provide a viable therapy following SCI. PMID- 17402854 TI - Spatial and temporal damage evolution after hemi-crush injury in rat spinal cord obtained by high b-value q-space diffusion magnetic resonance imaging. AB - Spinal cord injury (SCI) is a major cause of disability for many living persons. Therefore, several experimental models and handful of techniques were developed to study and characterize the damage evolution following SCI. In the present study, high b-value q-space diffusion-weighted imaging (DWI) was used to follow the spatial and temporal damage evolution in excised rat spinal cords following hemi-crush injury. The DWI results were correlated with behavioral testing. It was found that the damage depends, as expected, on the severity of the insult. Significant spontaneous recovery was observed, six weeks following the insult, only for the mild hemi-crush injury but not following the severe injury. The damage was found to be more severe in the area caudal to the trauma site as compared to the rostral section of the cord. PMID- 17402855 TI - Immediate damage to the blood-spinal cord barrier due to mechanical trauma. AB - Primary damage to the blood-spinal cord barrier (BSCB) is a nearly universal consequence of spinal cord injury that contributes significantly to the overall pathology, including the introduction of reactive species that induce cytotoxicity as well as secondary insults on the BSCB itself. We have characterized quantitatively the extent and severity of primary, physical disruption of the BSCB in adult rats 5 min after graded trauma induced with the Impactor weight-drop model of spinal cord contusion. Animals were injured by dropping a 10-g mass 12.5, 25, or 50 mm (n(level) = 8) on to the exposed mid thoracic spinal cord. The volume of extravasation of three markers of distinct size--fluorescently labeled hydrazide ( approximately 730 Da), fluorescently labeled bovine serum albumin ( approximately 70 kDa), and immunohistochemically labeled red blood cells ( approximately 5 microm in diameter)--were quantified in both the gray and white matter. The results indicate that spinal cord trauma causes immediate, non-specific vascular changes that are well-predicted by mechanical parameters. Extravasation volume increased significantly with increasing drop height and decreasing marker size. Extravasation volumes for all three markers were greater in gray matter than in white matter, and were better correlated to the rate of spinal cord compression than to the depth of spinal cord compression, which suggests that tissue-level strain rate effects contribute to primary spinal cord microvasculature pathology. The relationship between the response of the spinal cord and the injury pattern points towards opportunities to control the distribution and extent of injury patterns in animal models of spinal cord injury through a precise understanding of model and tissue biomechanics, as well as potential improvements in means of preventing spinal cord injury. PMID- 17402856 TI - Mesenchymal stem cells derived from peripheral blood protects against ischemia. AB - Intravenous delivery of mesenchymal stem cells (MSCs) prepared from bone marrow (BMSCs) reduces infarction volume and ameliorates functional deficits in a rat cerebral ischemia model. MSC-like multipotent precursor cells (PMSCs) have also been suggested to exist in peripheral blood. To test the hypothesis that treatment with PMSCs may have a therapeutic benefit in stroke, we compared the efficacy of systemic delivery of BMSCs and PMSCs. A permanent middle cerebral artery occlusion (MCAO) in rat was induced by intraluminal vascular occlusion with a microfilament. Rat BMSCs and PMSCs were prepared in culture and intravenously injected into the rats 6 h after MCAO. Lesion size was assessed at 6 h, and 1, 3, and 7 days using MR imaging and histology. The hemodynamic change of cerebral blood perfusion on stroke was assessed the same times using perfusion weighted image (PWI). Functional outcome was assessed using the treadmill stress test. Both BMSCs and PMSCs treated groups had reduced lesion volume, improved regional cerebral blood flow, and functional improvement compared to the control group. The therapeutic benefits of both MSC-treated groups were similar. These data suggest that PMSCs derived from peripheral blood could be an important cell source of cell therapy for stroke. PMID- 17402857 TI - Upregulation of vascular endothelial growth factor receptors Flt-1 and Flk-1 in rat hippocampus after transient forebrain ischemia. AB - This study characterizes the distribution of the two tyrosine kinase receptors for vascular endothelial growth factor (VEGF), Flt-1 and Flk-1, in the rat hippocampus following transient forebrain ischemia. The semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of Flt-1 and Flk-1 in hippocampal CA1 showed upregulation of these receptors following ischemic injury. Expression of Flt-1 and Flk-1 mRNA was restricted to neurons in the pyramidal cell and granule cell layers in control animals; however, upregulation was detected in activated glial cells and in the vascular endothelial cells rather than in neurons, in ischemic hippocampi. Most of the activated glial cells expressing Flt-1 and Flk-1 were reactive astrocytes, although some were microglial cells. The spatiotemporal expression of Flt-1 in the ischemic hippocampus mirrored that of Flk-1 expression. Expression of mRNA for both receptors was induced after 12 h, appeared to be increased progressively until 3 days when the highest expression was reached, and was sustained for more than 2 weeks. Flt-1 and Flk-1 immunoreactivity in the ischemic hippocampus matched the mRNA induction patterns except for a somewhat delayed onset. These data suggest that VEGF may be involved in the glial response via specific VEGF receptors in the rat hippocampus following transient forebrain ischemia. PMID- 17402858 TI - Diazoxide, as a postconditioning and delayed preconditioning trigger, increases HSP25 and HSP70 in the central nervous system following combined cerebral stroke and hemorrhagic shock. AB - Combined hemorrhagic shock (Shock) and unilateral common carotid artery occlusion (Stroke) results in a decrease of oxygen availability to peripheral tissues and organs and the central nervous system (CNS). A variety of biochemical processes ensue, including organ failure, cellular apoptosis, and necrosis. The present study used male, Sprague-Dawley rats to assess the impact of cerebral insult. Using heat-shock protein 25 and 70 (HSP25, HSP70) as biomarkers, measured 24 h after injury, we tested the hypothesis that pharmacological induction of preconditioning can offer cytoprotection from combined Stroke and Shock. The compound, diazoxide (DZ), is known to induce preconditioning through its effect as a mitochondrial potassium ATP (mK(ATP)) channel opener and succinate dehydrogenase inhibitor. When administered 24 h prior to Stroke and Shock (delayed preconditioning), DZ increased cerebral cortical and hippocampal levels of HSP25 and HSP70. A more clinically relevant treatment paradigm was tested, where DZ was administered after the induction of Stroke and Shock (postconditioning). When administered 60 min (but not 10 min) after the induction of Stroke and Shock, DZ significantly increased HSP25 and HSP70 expression in the ipsilateral cerebral cortex and hippocampus. Taken together, these results suggest that DZ treatment may be efficacious for CNS injury resulting from blood loss and anoxia from combined cerebral ischemia and hemorrhagic shock. "Postconditioning" triggered by DZ, immediately before resuscitation, is a potentially effective treatment for ischemia-reperfusion injury from combined Stroke and Shock. PMID- 17402859 TI - Ischemic vasoconstriction and tissue energy metabolism during global cerebral ischemia in gerbils. AB - Vasoconstriction is known to occur in cerebral arterioles during ischemia and considered to be distinct from vasospasm seen after subarachnoid hemorrhage. To elucidate the mechanism and functional significance underlying ischemic vasoconstriction, we investigated the relationship between arteriolar constriction and tissue energy metabolism during bilateral common carotid artery occlusion in gerbils. Using video microscopy and microspectroscopy, the arteriolar caliber, the total hemoglobin (Hb) content, and the redox state of cytochrome oxidase (cyt.aa3) were monitored in the cerebral cortex in vivo. After in situ freezing of the brain, adenine nucleotides, creatine phosphate (P-Cr), and lactate levels were analyzed using high-performance liquid chromatography in vitro. Tissue damage was also assessed immunohistochemically using antibodies against microtubule-associated proteins. There was a slight reduction of the diameter of pial arterioles during the initial 1 min of ischemia. A rapid decline of total Hb and reduction of cyt.aa3 were observed with rapid decreases of P-Cr and ATP in the cortical tissue during the initial 0.5 min, but all of them showed tendencies to return toward preischemic levels at 0.5-1 min. Beyond 1.5 min, extensive vasoconstriction occurred together with further decline of total Hb, reduction of cyt.aa3, and decreases of ATP and P-Cr. Neuronal damage developed in the cerebral cortex immunohistochemically beyond 3 min. The present investigation demonstrated two phases of vasoconstriction with the possibilities that the immediate vasoconstriction likely contributed to transient improvement of cortical oxygen/energy metabolism, and the second extensive vasoconstriction was an index of tissue energy failure and imminent neuronal damage. PMID- 17402860 TI - Mode changes of cerebrovascular pressure transmission induced by cerebral vasodilation. AB - Changes in the modes of cerebrovascular pressure transmission during cerebral vasodilation induced by hypercapnic challenge were examined as a means for developing the basis for a bedside method to evaluate regulation of cerebral blood flow. Recordings of arterial blood pressure (ABP) and intracranial pressure (ICP) obtained from a piglet preparation equipped with a cranial window were used to determine serial changes of the highest modal frequency (HMF) and dampening factor (DF) of a numerical system identification model of cerebrovascular pressure transmission. Resistance and compliance elements of a Windkessel model of ICP dynamics selected to provide the mathematical structure for the system identification modeling approach were also manipulated to obtain a match with HMF, DF, and the experimental and simulated recordings of ICP. During hypercapnic challenge, significant increases of ICP, pial arterial diameter (PAD) and partial pressure of arterial blood carbon dioxide increases, and a decrease of arterial pH were observed. Vasodilation changed the modes of the system identification model of cerebrovascular pressure transmission from a dominantly over-damped process to an under-damped one with a significant increase in HMF and decrease in DF. Simulations of the Windkessel circuit model required a decrease in the relative resistance and an increase in relative compliance of the arterial arteriolar vascular bed consistent with the observed increases in PAD induced by vasodilation. Evaluation of changes in the modes of cerebrovascular pressure transmission may provide means of assessing the state of cerebrovascular vasodilation and autoregulation of cerebral blood flow in the clinical setting. PMID- 17402861 TI - Thyroid hormone in biodegradable nerve guides stimulates sciatic nerve regeneration: a potential therapeutic approach for human peripheral nerve injuries. AB - It has been already demonstrated that thyroid hormone (T3) is one of the most important stimulating factors in peripheral nerve regeneration. We have recently shown that local administration of T3 in silicon tubes at the level of the transected rat sciatic nerve enhanced axonal regeneration and improved functional recovery. Silicon, however, cannot be used in humans because it causes a chronic inflammatory reaction. Therefore, in order to provide future clinical applications of thyroid hormone in human peripheral nerve lesions, we carried out comparative studies on the regeneration of transected rat sciatic nerve bridged either by biodegradable P(DLLA-(-CL) or by silicon nerve guides, both guides filled with either T3 or phosphate buffer. Our macroscopic observation revealed that 85% of the biodegradable guides allowed the expected regeneration of the transected sciatic nerve. The morphological, morphometric and electrophysiological analysis showed that T3 in biodegradable guides induces a significant increase in the number of myelinated regenerated axons (6862 +/- 1831 in control vs. 11799 +/- 1163 in T3-treated). Also, T3 skewed the diameter of myelinated axons toward larger values than in controls. Moreover, T3 increases the compound muscle action potential amplitude of the flexor and extensor muscles of the treated rats. This T3 stimulation in biodegradable guides was equally well to that obtained by using silicone guides. In conclusion, the administration of T3 in biodegradable guides significantly improves sciatic nerve regeneration, confirming the feasibility of our technique to provide a serious step towards future clinical application of T3 in human peripheral nerve injuries. PMID- 17402863 TI - The role of self-efficacy and referent specific social support in promoting rural adolescent girls' physical activity. AB - OBJECTIVE: To examine the role of social support (SS) and self-efficacy (SE) for physical activity (PA) in rural high school girls (N = 259, 15.5+1.2 yrs). METHODS: Using structural equation modeling, the relationships among PA, SS for PA from mother, father, and peers, and SE for overcoming barriers, seeking support, and resisting competing activities were tested. RESULTS: The effect of peer SS on PA was partially mediated through overcoming barriers. The effect of support seeking on PA was fully mediated through peer SS to overcoming barriers. Neither parent influenced PA. CONCLUSIONS: Peer SS and specific dimensions of SE should be considered in intervention studies. PMID- 17402864 TI - Alcohol consumption, medical conditions, and health behavior in older adults. AB - OBJECTIVES: To examine associations between drinking patterns, medical conditions, and behavioral health risks among older adults. METHODS: Analyses compared survey participants (health plan members ages 65 to 90, N = 6662) who drank moderately to those who drank over recommended limits or did not drink. RESULTS: Overlimit drinking was associated with smoking; not trying to eat low fat foods (in men), and lower BMI (in women). Predictors of not drinking during the prior 12 months included ethnicity, lower education, worse self-reported health, diabetes and heart problems. CONCLUSIONS: Significant relationships exist between health and alcohol consumption patterns, which vary by gender. PMID- 17402865 TI - Tobacco and other substance use among Alaska Native youth in western Alaska. AB - OBJECTIVE: To examine tobacco, alcohol, and other drug use rates among Alaska Native youth from western Alaska. METHODS: The sample consisted of 665 youth ages 6-18. RESULTS: Of children 6-10 years of age, 12% reported current use, and the prevalence rates increased with age. Females were significantly more likely than males to report tobacco use. The rates of alcohol and other drug use were very low. After adjusting for age and gender, significant correlates of tobacco use were maternal tobacco use during and after pregnancy. CONCLUSIONS: Expanded efforts are needed to address tobacco use among Alaska Native youth. PMID- 17402866 TI - Parental smoking, closeness to parents, and youth smoking. AB - OBJECTIVES: To examine parent closeness and its effect in predicting youth smoking when 0, 1, or 2 parents smoked. METHODS: Youth and parent smoking, closeness to parents, family structure, and gender and ethnicity among middle (n=17,468) and high school (n=5457) students were measured using a questionnaire. RESULTS: Number of parents smoking incrementally moderated the protective effect of all 4 measures of parent closeness, in predicting youth smoking. CONCLUSIONS: Addressing parent smoking and strengthening family relationships need significantly greater emphasis in interventions to further reduce youth smoking. PMID- 17402867 TI - Associations between depressed mood and clusters of health risk behaviors. AB - OBJECTIVES: To examine the association between depressed mood and clusters of health risk behaviors. METHODS: A nationally representative sample of adolescents (N=15,214) was utilized to construct 10 pseudocontinuous health risk behaviors. Cluster analysis was performed to group adolescents, and subsequent multivariable logistic models were created. RESULTS: Compared to non-risk takers, belonging to risk clusters significantly increased the odds of reporting depressed mood. African Americans in high-risk clusters appeared to be more vulnerable to depressed mood, when compared to remaining racial groups. CONCLUSIONS: Results suggest that adolescents engaging in multiple health-risk behaviors do so in the context of depressed mood. PMID- 17402868 TI - Balancing quality and cost for Adult Tobacco Telephone Surveys. AB - OBJECTIVES: To demonstrate the ability to cost-effectively coordinate Adult Tobacco Survey stakeholder interests while reducing the risk of potential bias. METHODS: Key smoking indicators were compared across 2 surveys and analyzed based on modifications to calling protocols. RESULTS: Mixed results were found when comparing smoking rates across 2 surveys, by early, mid, and late respondents, and by the number of rufusals. Significant cost sayings can be obtained by reducing the number of telephone call attempts. CONCLUSIONS: Few significant differences may encourage reductions in protocol, but this must be weighed against the possibility of cost-saving measures resulting in biased estimates. PMID- 17402869 TI - Differences in perceived implementation of a standard versus peer-led interactive substance abuse prevention program. AB - OBJECTIVE: To assess perceived implementation of 2 substance-abuse prevention programs: a standard one and a peer-led interactive one. METHODS: Data from 16 health educators were collected after 504 classroom sessions, 63 of which were observed by 24 monitors. RESULTS: In the interactive program, health educators (HEs) followed the curriculum less closely, reported less favorable classroom processes and less off-task talking than in the standard one. CONCLUSIONS: These data indicate that an interactive substance-abuse prevention program does not necessarily entail more off-task discussion but also does not necessarily guarantee more favorable program implementation. PMID- 17402870 TI - Computers as a formal continuing education tool: moving beyond intention. AB - OBJECTIVE: To identify factors related to health promotion professionals' decisions to use computer-mediated instruction for continuing education (CMI-CE). METHODS: Employing a cross-sectional survey design, data were collected from 500 respondents using an online survey. RESULTS: Among respondents having positive intentions toward CMI-CE, characteristics distinguishing between individuals with positive and negative CMI-CE behavior included perceived behavioral control, license/ certification, lack of programs and relevant program topics, and availability of technical support. CONCLUSIONS: Health promotion professionals have positive intentions toward CMI-CE, and introducing relevant programs accompanied by strong technical support teams will help move them beyond intention and into action. PMID- 17402871 TI - HIV/AIDS-related knowledge and attitudes of Palestinian women in the Occupied Palestinian Territory. AB - OBJECTIVE: To explore Palestinian women's knowledge, perceptions, and attitudes towards HIV/ AIDS. METHODS: Data used in this study were collected as a part of the Demographic and Health Survey (DHS) 2004 in the Occupied Palestinian Territory (OPT). RESULTS: Most of the women in the OPT (93.4%) had heard of AIDS. They had fair general knowledge of HIV/AIDS prevention methods and transmission. TV was the main source of information about HIV/AIDS. Among the determinants of HIV/AIDS knowledge identified by this study were education, region, and locality. CONCLUSION: A research-informed policy concerning HIV/AIDS prevention should be formulated. PMID- 17402873 TI - [Home hemodialysis: a possible and necessary alternative]. PMID- 17402872 TI - [Lights and shades in the lupus nephritis treatment]. PMID- 17402874 TI - [PPAR-gamma, arterial hypertension and the kidney]. PMID- 17402875 TI - [Intravenous cyclophosphamide in lupus nephritis: twenty years reducing dose]. AB - The prognosis for patients with proliferative glomerulonephritis associated with systemic lupus erythematosus has dramatically improved over recent decades. We review our experience with intermittent pulse therapy with intravenous cyclophosphamide (IC) in 97 patients (75 female) aged over 20 years. The series was divided into three groups. Group A (n=39) received monthly IC pulses (begin 1 g) for up to 24 months between 1985-1991. Group B (n=47) received monthly IC pulses (1 g) for six months with additional quarterly doses for a maximum of 18 months, depending on the therapeutic response (from 1991). From 1999, Group C (n=11) patients were treated with low-dose IC (3 g in three months) followed by azathioprine (2 mg/kg) or mycophenolate mofetil (1.5-2.0 g/day) for 12-18 months. The total IC doses (g) administered were: Group A, 15.1+/-9.0; Group B, 8.5+/ 3.5; and Group C, 3.0+/-0.0. These figures show the trend progressive reduction in exposure to IC. Overall, treatment with the different IC regimens achieved satisfactory control of lupus nephritis in 76% of the patients. Comparison of the values at baseline and after 24 months showed that the serum creatinine (mg/dl) fell in Group A from 1.77+/-1.06 to 1.09+/-0.63, in Group B from 1.22+/- 0.85 to 0.95+/- 0.45, and in Group C from 0.90+/-0.23 to 1.17+/-0.54 (p<0.05). In the same period, proteinuria (g/day) fell in Group A from 6.19+/-4.31 to 0.79+/-1.76, in Group B from 4.43+/- 3.17 to 2.08+/-3.65, and in Group C from 5.43+/- 3.37 to 3.22+/-4.00 (p=0.05). There was not differences between the three groups in both variables. The adverse effects were mainly viral and bacterial infections, with no intergroup differences. Avascular osteonecrosis requiring hip replacement and early menopause were more frequent in Group A. Nine patients died, seven due to cardiovascular causes and two with infection. No differences were detected between the three groups when analyzing the overall patient survival at 5, 10 and 15 years (95%, 92%, and 84%, respectively). The likelihood of maintaining serum creatinine within normal ranges or less than twice the baseline range was similar in the three groups at 5, 10 and 15 years (92%, 72% and 66%, respectively). There were 47 episodes of relapse, with no differences between the three groups. In Summary, treatment with different regimens of intermittent IC is relatively safe and efficient to control the disease and lupus nephritis in SLE patients even with progressively smaller doses. The price paid concerned infectious complications, and bone and ovarian toxicity. New alternatives should at least maintain the same efficacy, but with fewer adverse effects and relapses. PMID- 17402876 TI - [Glutamin concentrations in patients treated with cisplatin have a predicting value of renal failure development]. AB - Glutamine is an amino acid (AA) whose mission is carrying nitrogen. Abnormalities in protein and AA metabolism has been observed in patients with acute renal failure. Several clinical trials had showed abnormalities in plasma AA and its elimination in uremic environment. Moreover, renal failure disturbs hepato splachnic circulation of glutamin, tyrosine and proline. Cisplatin is a key drug in the chemotherapy pharmacology, and the nephrotoxicity due to tubular injury, is one of its most important side effects, which sometimes is irreversible and leads to substitutive renal treatment. The goal of this work is to find predictive factors of renal failure secondary to cisplatin. Fifty four patients treated with cisplatin were studied. The plasma AA concentration and another plasmatic and urine parameters were measured in three different days after each pulse of chemotherapy. Plasma AA modifications through the pulses and reabsortion percentages of everyone were analysed too. Significant differences were observed in 13 AA reabsortion percentages and 16 plasmatic concentration. Glutamin concentration through the pulses was higher in 13 patients, (24%) who presented renal failure (Plasmatic glutamin concentration higher than 1000 mM/L at the third day after cisplatin administration was highly predictive value about getting renal failure, with significant difference from patients with o normal renal function. Others parameters analysed did not rise significant predictive values, so as it was not found relation between hyperaminoaciduria and renal function. It is concluded that cisplatin leads to renal failure in a 24% of this patients. Glutamin, concentrations higher than 1000 mM/L at the third day after cisplatin administration have a high predictive value about getting renal failure; so, it is suggested this could be a early marker of cisplatin nephrotoxicity before the serum creatinine is elevated, in order to get an early and suitable treatment of it. PMID- 17402877 TI - [Usefulness of oral glucose tolerance test (OGTT) in output patients with advanced chronic renal failure (CRF)]. AB - A pathological Oral Glucose Tolerance test (OGTT) is early marker of peripheral insulin resistance. Nevertheless, its utility in nondiabetic patients with CRF stage IV-V is undetermined. AIM: We wanted to detect, in a population of non diabetic patients with CRF, the presence of carbohydrates metabolism anomalies, by means of the OGTT and to relate it with metabolic, anthropometric, cardiovascular parameters and renal function. We studied 45 non diabetic patients with advanced CRF (stage IV-V), 26 men, mean age 66.5 years, with average Cockroft-Gault of 23.6 ml/min. We measured weight, height, waist and BMI. BIOCHEMICAL: glucose, insulin, OGTT, C peptide, lipid profile, HbA1C and Hto. Cardiovascular comorbidity, mean proteinuria and systolic and diastolic blood pressure (6 months pre and post analytical measure) were measured. Pulse pressure was also calculated. RESULTS: 47% of the patients presented normal fasting glucose, whereas 53% had isolated impaired fasting glucose (IFG). After the OGTT, 36% of the patients presented impaired glucose tolerance (IGT) and 14% diabetes (>200 mg/dl). Of the patients with normal fasting glucose, 38% had IGT after OGTT and 5% diabetes. Patients with abnormal OGTT were older (71+/-13.6 versus 60+/- 18.8 years, p=0.03), had greater HbA1C (5.6+/-0.5 versus 5.2+/-0.3%, p=0.02), total cholesterol (193+/-37.7 versus 169.8+/-44.9 mg/dl, p=0.03), pulse pressure (63.4+/- 14.5 versus 52.3+/-9.7 mmHg, p=0.0001) and greater prevalence of ischemic heart disease (28% versus 5%, p=0.05). Creatinine Clerance negatively correlated with the OGTT (r=-0.39, p=0.01) and plasma creatinine positively with fasting insulin (r=0.33, p=0.02) and C-peptide (r=0.42, p=0.006). Urinary Proteins were correlated with fasting glucose (r=0.30, p=0.04), C-peptide (r=0.52, p=0.001), triglycerides (r=0.36, p=0.01) and with the HOMA-IR index (r=0.30, p=0.05). CONCLUSION: Fasting Glucose did not predict OGTT results in patients with CRF. For this reason, we think that the OGTT can be very usefull tool to identify states of "prediabetes" and diabetes in patients with CRF, specially in those whose present an elevated Pulse Pressure, age greater than 65 years, hyperlipidaemia and HbA1C above 5.2%. The early detection of these metabolic anomalies, may lead to intensify dietetic and pharmacological measures directed to delay or to attenuate the appearance of diabetes and its serious complications in a population in which the cardiovascular risks factors are very elevated. PMID- 17402878 TI - [Nutrient intake and eating habits in hemodialysis patients: comparison with a model based on mediterranean diet]. AB - OBJECTIVES: Inadequate nutrient intake seems to be one of the most important cause of malnutrition in hemodialysis patients. The purpose of this study was to analyse their nutrient intake and eating habits, comparing food groups' intake with standard Mediterranean diet values (Healthy Diet Guide 2004, Nutrition Community Spanish Society). MATERIAL AND METHODS: There were 28 stable hemodialysis (HD) patients, 15 males and 13 females, mean age 62,9 +/- 16 years. Dietary evaluation was based on 7-day dietary recalls conducted by a single observer. We compare nutrients intake with recommended hemodialysis intake and we contrast food groups consumption with the theoretical ideal based on Mediterranean diet. RESULTS: The protein intake was 1,33 +/- 0,2 g/kg/day and the energy intake 29,5 +/- 2,1 kcal/kg/day. Carbohydrates accounted 43,1% of energy intake, proteins 19% and lipids 37,9% (55,5% monounsaturated fatty acids, 16,4% polyunsaturated fatty acids and 28,1% saturated fatty acids). Complex carbohydrates (potatoes, cereals, vegetables, fruits) and olive oil consumption was lower than that recommended to the Spanish healthy population and to the chronic hemodialysis patients. The animal protein intake (meat, fish, eggs) was correct, although excessive in red and processed meats. Results: Potatoes and cereals recommended frequency (RF) 4-6 portions/day, HD patients frequency (HDF) 4,1 portions/day; vegetables RF > 2 portions/day, HDF 1,2; fruits RF > 3 portions/day, HDF 1,3; olive oil RF 3-6 portions/day, HDF 1,5; Fish RF 3-4 portions/week, HDF 4,2; White meat RF 3-4 portions/week, HDF 1,5; Poultry RF 3-4 portions/week, HDF 2,3; Eggs RF 3-4 portions/week, HDF 3,6; Pulses RF 3-4 portions/week, HDF 1,7; Nuts RF 3-7 portions/week, HDF 0; Red meat RF occasionally, HDF 4,8 portions/week; Processed meats RF occasionally, HDF 4,6 portions/week; Sweets, snacks, soft drinks RF occasionally, HDF 1,7 portions/week; Butter, margarine, processed bakery products, biscuits RF occasionally , HDF 0,5 portions/week. CONCLUSIONS: Nutritional abnormalities are frequently found even in apparently stable patients on chronic hemodialysis. Caloric rather than protein undernutrition is the major abnormality. Inadequate caloric intake (< 35 kcal/kg/day) can lead to a negative nitrogen balance. Their eating habits are healthy and natural, but there is a deficit in slowly absorbed carbohydrates and olive oil intake (with caloric intake reduction), and an excessive consumption of red and processed meats (with saturated fats increase). The individual correction of these dietary patterns could reduce the saturated fats and increase the energy intake, obtaining a balanced diet integrated into our geographic region and culture. PMID- 17402879 TI - [Factors which influence phosphorus removal in hemodialysis]. AB - BACKGROUND: The sustained elevation of phosphorous among patients with end-stage renal failure is associated with elevated mortality rates. Phosphate binding agents are usually necessary to control serum phosphate levels. Phosphate removal during dialysis is limited largely due to the intracellular location of most inorganic phosphorous. The membrane surface, the frequency and the duration of therapy have proved to be very important factors in the serum phosphate control. THE AIM of our work is to investigate the influence on phosphate removal of factors that normally participate in the haemodialysis session: Plasma phosphate level (Php), treatment duration, membrane surface, high or low-flux membranes, the vascular access, dialysate flux , the volume of blood passing through the dialyzer (L) in each dialysis session and the blood flow during the first hour of dialysis. On 16 patients, we also had the possibility of comparing phosphate removal with 1.8 m(2) high-flux haemodialysis, 1.8 m(2) on-line hemodiafiltration and the on-line technique with the new Helixone dialyzer Fresenius Fx100. METHODS: 108 haemodialysis patients, 62% men, 38% women aged 21-82 years (61+/ 14;mean+/-sem),) were selected for the study. Mean treatment time 4.14+/-0.41 hours (range 3.5-5 hours). The vascular access was an arterio-venous fistula in eighty five (78%) and a double lumen tunnelled catheter 23 (22%). Patients were studied under their normal every day conditions. High-flux membrane was used by 31 (30%) patients and low-flux membrane by 77 (70%). Membrane surface was: 1.7 m2:17 (16%); 1.8 m2:77 (71%); 2,1 m2:14 (13%). Dialysate flux was: 500 ml/min. 55 patients; 700 ml/min: 53 patients. In 16 out of 108 patients we had the possibility of using on-line hemodiafiltration with ultrapure bicarbonate buffered dialysate. Phosphate mass removal (MPO4) was calculated using the formula:MPO4=0.1 t-17+50 Cds 60+11Cb 60 (1), where t is treatment time in minutes, Cds60 and Cb60 are phosphate concentrations in dialysate and plasma measured at 60 min from the beginning of hemodialysis in mg/dl, and MPO4 is the estimated phosphate removed in mg/treatment. RESULTS: We found a good correlation between phosphate removal and serum phosphate levels (p=0.01), but not with the membrane surface or treatment duration. Phosphate removal was 640+/-180 mg/session with low-flux membrane and 700+/-170 mg/session with high-flux membrane (p=0.280). The MPO4 was 720+/-190 mg/treatment in patients with a AV fistula and 620+/-180 in patients with a tunnelled catheter (p=0.023). We found a good correlation between phosphate removal and the volume of blood (L) that passed the dialyzer in each session (r=0.001) but we did not find a correlation between phosphate removal and KT/Vurea, the dialysate flux or the ultra filtration. On-line technique did not increased the MPO4(733+/-280 mg, p=0.383). The on-line technique with the new dialyzer (Fresenius Fx100), increased the phosphate removal to 759+/-199 mg/session (p=0.057). CONCLUSION: Phosphate removal during dialysis is influenced by Plasma phosphate levels, the volume of blood that passed the dialyzer and the vascular access. Uniformity on time and membrane surface could explain the abs cense of influence in our case. The ultra filtration, dialysate flux, membrane permeability or on-line hemodiafiltration does not influence the phosphate removal. The new membrane helixone with 2,1 m2 (Fresenius Fx100) increases phosphate removal probably because the membrane surface is higher. PMID- 17402880 TI - [Hospital admisions and consultations in a dialysis population]. AB - The hemodialysis (HD) population continues growing and aging, reason why they are required to adapt the human, economic and materials resources to be taken care of. OBJECTIVE: To analyze the distribution of the consultations and the times of admissions in the different Hospital services of which an peripheral arranged HD Center depends. METHOD: We analyzed all the consultations to the Hospital Urgencies Departments and Hospitable admissions of all the patients who received HD in the arranged Center Ponfedial, (sanitary area Ponferrada of 150,000 inhabitants) in period 01/12/02 to the 31/05/05. RESULTS: We prospectively examined a population of 87 HD patients (71.0+/-12.5 years, 77.9 % male, 31.4% diabetics) time in HD 21.0+/-22.7 months. They were a total of 286 (9.5/month) consultations in the Service of Urgencies (2.31 consult/patient/year), 37.4% of them did not motivate admissions. The most frequent consultation (28,9%) was the complications of vascular access (70,9% of them required positioning a transitory catheter; 1.1 complication of catheter/months, 1 permanent catheter is placed/1.2 months and 1 fistulography/1.2 months), cardiac causes 10.3%, acute pulmonar edema: 9.3%, infectious: 8.4% and traumatologic: 8.4%. There were 179 admissions (5.9/month; 1.44 hospitalizations/patient/year; 4.0+/-14.2 days of admissions): cardiovascular cause: 37.4%, by vascular access: 18.4%, infectious disease: 17.0%, digestive: 6.7%, traumatologic: 3.9% and by other causes: 11.0%. The cardiovascular group takes 46.5% (60.8% of them by vascular peripheral disease) of the total of days of hospitalizations, infectious 16.7%, neoplasic 8.4% and vascular access 7.6%. The neoplasic causes were the greater average with 36.0+/ 21.6 days of hospitalization, by amputations 23.0+/-38.1 days, by stroke 14.5+/ 10.6 days, infected ulcers of lower extremities members 10.0+/-19.0, other vascular treatments 10.0+/-4.7, ischemic heart disease 8.5+/-20.3 and heart failure 7.5+/-5.2 days. Our prevalent population of 49.5 patients needed 3 to 7 hospitable beds, 0.33 temporal catheter, 1 tunneled permanent catheter and 1 fistulography per month. CONCLUSIONS: The cardiovascular causes, the accesses and the infectious ones are the main causes of consultations, hospital admissions and time of hospitalizations. PMID- 17402881 TI - [Seasonal perodicity in vascular access thrombosis for haemodialysis]. AB - Different studies have shown that some clinical events, particularly cardiovascular and thrombotic events, show a regularity in its appearance. The aim of our study was to analyse the possible existence of seasonal periodicity in the incidence of the vascular access thrombosis in patients on chronic haemodialysis. Prospectively, we collected information of 164 patients with 250 episodes of vascular access thrombosis referred to our hospital from january 1995 to december 1999. An ANOVA test for comparison of the means, and a time series analysis were performed. During the five year study the consecutive number of thrombosis were 43, 57, 55, 59 and 36. When the different seasons were analysed, the cumulative number of events in summer during the study period were 91, a significant increase compared to spring, autumn, and winter (54, 54, and 51, respectively; p<0.001). Time series analysis confirmed that thrombolic events during summer showed an increased incidence over the mean (p<0.001), and it occurred every year. The same results were obtained when the PTFE grafts were analyzed separetely (july RR 2.62, p=0.002; august, RR 2.37, p=0.04), but not with the arteriovenous fistulae. In conclusion, this study showed a seasonal periodicity of vascular access thrombosis, with a PTFE graft. Although the causes were unknown, these data alert us on the convenience of an increased attention to the vascular access during the summer months in order to prevent its thrombosis. PMID- 17402882 TI - [Control of the dialysis dose by ionic dialysance and bioimpedance]. AB - INTRODUCTION: The ionic dialysance monitor allows an automated measure of Kt in each dialysis session. Bioelectrical impedance analysis (BIA) determines the total body water which it is equivalent to the urea volume of distribution (V). If the Kt, determined by ionic dialysance, is divided by the V, estimated by bioelectrical impedance, a Kt/V at the end of dialysis session (Kt/VDiBi) is obtained. AIM OF THE STUDY: To evaluate the agreement between the Kt/VDiBi and the Kt/V obtained by two simplified formulas: the monocompartimental (Kt/Vm) and the equilibrated (Kt/Ve) Daugirdas equations. METHODS: The Kt/VDiBi, the Kt/Vm and the Kt/Ve were determined in 38 hemodialysis patients (27 males and 11 females) in the same hemodialysis session. The patients were on dialysis three times a week for 3.5 to 4 hours. The V was determined by monofrequency bioelectrical impedance (50 kHz) at the end of the dialysis session. RESULTS: The Kt/VDiBi, Kt/Vm and Kt/Ve were 1.29+/-0.26, 1.54+/-0.29 and 1.36+/-0.25, respectively (p<0.001 between the Kt/VDiBi and the KtVm, and p<0.001 between the KtV/DiBi and the Kt/Ve). The intraclass correlation coefficient showed better concordance between the KtV/DiBi and the Kt/Ve (coefficient 0.88) than between the Kt/VDiBi and the KtVm (coefficient 0.65). The relative difference of the Kt/VDiBi was 8.3+/-6.4% with respect to the Kt/Ve and 18.4+/-7.8 % with respect to the Kt/Vm (p<0.001). The relative difference between the Kt/VDiBi and the Kt/Ve was lower than 15% in the 84% of the patients and lower than 10% in the 64% of the patients. CONCLUSIONS: If the V obtained by bioelectrical impedance analysis is included in the ionic dialysance monitor, we can obtain a Kt/V for each patient in real time, which is similar to the equilibrated Kt/V obtained from the Daugirdas equation. PMID- 17402883 TI - [Parathyroid cancer in a patient with previous history of hypernephroma: a clinical case]. AB - We report the clinical case of a 55 year-old male patient, with a previous history of nephrectomy by hypernephroma sixteen years ago, first presenting hypercalcemia and rising of intact parathyroid hormone (iPTH) levels. A localization study revealed an intrathyroid nodule with cystic appearance. After undergoing a hemi-thyroidectomy, the patient is diagnosed with parathyroid carcinoma. This article analyzes previously published cases presenting parathyroidal pathologies associated with hypernephroma. A broader differential diagnosis--including the screening of parathyroidal pathologies should be considered in patients with hypercalcemia and hypernephroma. PMID- 17402884 TI - [Good virological response to pegylated interferon alfa monotherapy of chronic hepatitis C infection in hemodialysis patient]. AB - Liver disease caused by hepatitis C virus infection is associated to significant morbidity and mortality among patient with end stage renal disease on maintenance hemodialysis (HD). Therapy in these patients consists of Interferon, preferably pegylated Interferon (pIFN), thus Ribavirin (RBV) is not recommended for patients with impaired renal function, outside its use in controlled trials. We report a case of 35 years young woman on HD treatment, renal transplantation candidate with chronic hepatitis C virus infection, HCV RNA positive (by PCR), genotype 3a, moderate viral load, light increase of aminotransferases. Pegylated Interferon alfa-2a (135 mcg/weekly/SC) was initiated. She achieved HVC RNA negative within 12 weeks, following with pINF as monotherapy to complete 24 weeks (6 months). Sustained virologic response persisted to 24 and 48 weeks. Most important side effects were light detriment of anemia, moderate neutropenia and thombocytopenia, transitory elevation of transaminases and "flu-like" syndrome. Adverse events were well tolerated with total compliance with pIFN dose, no requiring reduce or stop the treatment. These findings confirm that hemodialysis patients with chronic hepatitis C respond well to pegylated IFN monotherapy and a long-term sustained virologic response is achieved, appears to be better tolerated with less side effects, so combination therapy with pINF plus ribavirin is not necessary in all cases. PMID- 17402885 TI - [Prototheca Wickerhami peritonitis in patients on peritoneal dialysis]. AB - Prototheca spp exits throughout nature but only infrequently cause infection in humans. Most of the cases of human infection have involved the skin or subcutaneous tissues, but there have been reports of rare cases of protothecosis of the urinary tract, and disseminated disease. The species most commonly isolated is Prototheca wickerhamii. Few cases of peritonitis due to P. wickerhamii in peritoneal dialysis had been reported. We report a successful treatment of Prototheca peritonitis complicating peritoneal dialysis with amphotericin, itraconazole and removal of the catheter. PMID- 17402886 TI - [Nephrotic syndrome secondary to focal and segmental glomerulosclerosis in a patient with alveolar sarcoidosis]. PMID- 17402887 TI - [Deterioration of renal function associated with abdominal pressure increase]. PMID- 17402888 TI - [Lupus nephritis and hypothyroidism]. AB - Thyroid diseases could be associated with several renal alterations. In the present report we describe a patient with SLE with inactive renal involvement and symptomatic hypothyroidism that developed a clinical picture similar to lupus flare. PMID- 17402889 TI - [Diabetic nephropathy. Pending assignment]. PMID- 17402890 TI - [Klebsiella Pneumoniae pericarditis in patient with systemic lupus erythematosus and kidney failure]. PMID- 17402891 TI - [Emphysematous pyelonephritis with perirenal abscess cured with conservative medical treatment]. PMID- 17402892 TI - [Complete remission of nephrotic syndrome with methylprednisolone pulses in an adult with Schonlein-Henoch purpura]. AB - Henoch-Schonlein purpura is a systemic vasculitis that occurs most frequently in childhood. Massive proteinuria, renal impairement at onset and histologic severity in renal biopsy are considered the main risk factors for deterioration of renal function at long-term. We report a 24 years-old woman with Henoch Schonlein purpura who developped a severe nephrotic syndrome with microhematuria and normal renal function. Renal biopsy showed a diffuse endocapillary proliferative glomerulonephritis with less than 50% crescents (type IIIB of ISKDC classification). As their potential bad prognosis we decided to treat with methyl prednisolone pulses (3 x 500 mg in months 0, 3 and 5) accompanied by maintenance treatment with prednisone (0,5 mg/kg/every other day) for 9 months. We observed with this protocol complete remission of nephritis with preservation of renal function. PMID- 17402894 TI - Quality junior doctor training, improved workforce outcomes and patient safety. AB - The intern year is a critical part of medical education and pivotal in provision of health services, especially in tertiary facilities. Training must be integrated with health service needs, as our health service is not well served if junior doctor training creates confidence without competence. PMID- 17402895 TI - Redesigning models of patient care delivery and organisation: building collegial generosity in response to workplace challenges. AB - This case study describes the New South Wales Nursing and Midwifery Office (NaMO) Models of Care Project, a project designed to identify, encourage and disseminate innovations in nursing care organisation and delivery. The project is a 4-year action research project, using a range of interactive engagements including workshops, seminars, questionnaires and websites to achieve the goals. This case study briefly describes the main stimuli for review and redesign of models of care identified through analysis of the clinicians' presentations, and explores the range of responses to the workplace challenges. PMID- 17402896 TI - Anaesthesia underpins acute patient care in hospitals. AB - The Australian and New Zealand College of Anaesthetists (ANZCA) carried out a review of the roles of anaesthetists in providing acute care services in both public and private hospitals in Europe, North America and South-East Asia. As a result, ANZCA revised its education and training program and its processes relating to overseas-trained specialists. The new training program, introduced in 2004, formed the basis for submissions to the Australian Medical Council, and the Australian Competition and Consumer Commission/Australian Health Workforce Officials' Committee review of medical colleges. A revised continuing professional development program will be in place in 2007. Anaesthetists in Australia and New Zealand play a pivotal role in providing services in both public and private hospitals, as well as supporting intensive care medicine, pain medicine and hyperbaric medicine. Anaesthesia allows surgery, obstetrics, procedural medicine and interventional medical imaging to function optimally, by ensuring that the patient journey is safe and has high quality care. Specialist anaesthetists in Australia now exceed Australian Medical Workforce Advisory Committee recommendations. PMID- 17402897 TI - Changing the face of mental health care through needs-based planning. AB - Mental disorders contribute to the burden of human disease. The National Survey of Mental Health and Wellbeing revealed low participation in treatment. The Tolkien II report provided evidence that a mental health service that utilised needs-based stepped care was likely to be effective and affordable to the point that a 30% increase in budget would treat 60% more people and produce a 90% increase in health gain. Five priorities were identified: Solve the crisis in psychosis by providing more step-down beds for people with schizophrenia who need long-term accommodation. Educate the workforce by providing a nationwide web based basic curriculum. Use clinician guided, step-down web-based therapy for patients who are mild or moderate, and web-based education to enhance clinical treatment for patients who are more severe. Educate patients and their families about treatments that work and about lifestyle changes that facilitate these treatments. Reduce the onset of common mental disorders by using proven web-based prevention programs in schools. With resources such as these in place, changing the face of mental health care might just be within our reach. PMID- 17402898 TI - Managing the interface between acute care and rehabilitation - can utilisation review assist? AB - AIMS AND METHODS: We piloted the InterQual Criteria tool in a large regional acute hospital in NSW to determine the utility of this tool in the Australian context. In particular to compare the current "gold standard" of physician assessment for the selection of patients for rehabilitation and the timing of transfer, with the guidance provided by the tool. Consecutive acute care patients with a diagnosis of stroke, hip fracture or amputation, and patients referred for rehabilitation assessment, were followed using the InterQual Criteria. RESULTS: Results on 242 acute episodes, representing 2698 days in acute care, were analysed. In accordance with overseas studies, we found that high levels of inappropriate days of stay in acute care were suggested by the tool. Using the InterQual Criteria almost all patients were deemed appropriate for transfer to rehabilitation much earlier than current practice. CONCLUSION: We conclude that the InterQual Criteria may have a useful role in patient selection for rehabilitation, in facilitating the transfer of patients from acute to subacute care, and in improving patient flow within acute care. The reasons for the variation between the results obtained from the tool and current clinical practice requires further investigation, and may indicate a lack of validity of the tool in the Australian setting, inefficiencies in processes of acute care, or the lack of suitable alternative care settings or level of support available in these settings. PMID- 17402899 TI - Community consultation and engagement in health care reform. AB - In advocating for significant reform of the health care system, the Australian Health Care Reform Alliance (AHCRA) supports a process of citizen engagement that will allow the wider community to have a say in the future direction of their health care system. Models that have engaged community opinions have been successful overseas, and this article calls for similar processes in Australia. PMID- 17402900 TI - A new approach to clinical governance in Queensland. AB - Clinical governance approaches in Queensland health were trenchantly criticised in 2005 by two external reviews. In designing the new approach to clinical governance it was recognised that clinical governance should not be seen as only being about traditional safety and quality policies. A range of levers and policy instruments have been used in Queensland health to effect a new approach to clinical governance. PMID- 17402901 TI - Medical role substitutions and delegations - overcoming the fear. AB - This paper outlines the increasing need for new health care practitioners to work under delegation or as substitutes for medical practitioners to resolve the Australian health workforce crisis. The personal experience of the author and common issues for medical practitioners related to the introduction of these new roles are discussed. PMID- 17402902 TI - Teaching, learning and research: essential elements of health care and the next Australian Health Care Agreements. AB - Australian Health Care Agreements must set out the roles and responsibilities of the Australian, state and territory governments in ensuring the provision of appropriate clinical placements for the additional medical student allocation. The roles of universities and public and private health agencies must also be specified. PMID- 17402903 TI - The impact of the pathology workforce crisis on acute health care. AB - There is much talk of the crisis in medical workforce. This paper outlines the huge problem facing pathology services in Australia and the impact this will have on the delivery of health care services. PMID- 17402904 TI - The AROC Annual Report: the state of rehabilitation in Australia 2005. AB - This is the inaugural comprehensive annual report that describes patients discharged from subacute inpatient rehabilitation programs provided by facilities that are members of the Australasian Rehabilitation Outcomes Centre (AROC). Collection of a standardised dataset has enabled the provision of a national benchmarking system, which in turn has led to an improved understanding of factors that influence rehabilitation outcomes and costs, and therefore performance of the sector. PMID- 17402905 TI - Hospitals then and now: changes since the start of Medicare. AB - Australia's current hospital system is very different to the one that existed in 1982-83, the last full year before the introduction of Medicare. Capped budgets, new technology and rising expectations have seen the public sector lose ground to the private sector, particularly in terms of elective admissions and surgical episodes. In comparing the situation in 2004-05 with that in 1982-83, this paper focuses on hospital efficiency, casemix and the public-private balance. PMID- 17402906 TI - Connecting funds with quality outcomes in health care: a blueprint for a Clinical Practice Improvement Payment. AB - There are significant geographic variations in the quality of health care, often with substantial gaps between what is known to be achievable and what is actually achieved in practice. This is a global problem that has persisted for many years despite a variety of conventional quality improvement initiatives. Attention has therefore recently turned to realignment of funding with specified levels of desired quality of care as an alternative. This paper outlines one approach that will be introduced as a pilot in Queensland. PMID- 17402907 TI - Mature use of casemix - are we there yet? AB - In this paper we consider the progress made in using casemix in Australia. We argue that while the casemix infrastructure has been highly developed and the casemix-based funding systems of some states are mature, there is still more development needed to use the data for clinical questions such as quality improvement activities. Further research is needed to establish what is needed to describe the impact of casemix on clinicians and hospitals in a context of increasing accountability and transparency and where questions of efficiency cannot be ignored. PMID- 17402908 TI - The development of Version 2 of the AN-SNAP casemix classification system. AB - This paper presents the results of a recent review of the Australian National Sub acute and Non-acute Patient (AN-SNAP) classification system. The AN-SNAP system was developed by the Centre for Health Service Development, University of Wollongong in 1997. The review was conducted between August 2005 and September 2006. Four clinical sub-committees comprising more than 50 clinicians from sub acute services across New South Wales as well as representatives from Queensland and the Australian Capital Territory were established to develop a set of proposals to be considered for incorporation into Version 2 of the classification. It is proposed that the final AN-SNAP Version 2 classification will be available for implementation from 1 July 2007. PMID- 17402909 TI - Supporting work practices, improving patient flow and monitoring performance using a clinical information management system. AB - Providing information technology solutions to clinicians to support their work practices benefits clinicians, administrators and patients. We present our 8-year experience with an inexpensive information management system which provides clinical and business process support for clinicians and bed managers. The system has been used by an area rehabilitation and aged care service to manage inpatient consultations and patient flow across nine hospitals. Performance monitoring of the time from referral to consultation, the number, type and outcome of consultations, and the time taken to access a rehabilitation or subacute bed is also provided. Read-only access to the system for clinicians and bed managers outside the rehabilitation and aged care service allows greater transparency. PMID- 17402910 TI - Quali-quantitative analysis: a new model for evaluation of unusual cases in hospital performance? AB - This paper aims to provide hospital administrators and others making decisions about hospital error funding, as well as researchers, with information about what good hospital error research looks like. It offers a selective review of how the error literature has approached hospital error, which is used to develop five criteria for sound hospital error research. It also explores the potential for better hospital error research of quali-quantitative analysis (QQA), an innovative social sciences research method. In a context in which other methodologies all have their shortcomings, QQA appears to go some way toward meeting the five criteria for sound hospital error research. Ideally, QQA would be used in combination with other approaches to answer the kinds of questions that are important to hospital administrators when they are faced with high stakes error evaluation situations. PMID- 17402911 TI - Workforce development: planning what you need starts with knowing what you have. AB - Health workforce planning and research occurs at both national and state/territory levels, but identifying current workforce availability and future workforce need is more problematic at a regional level. We report on the practical approach to workforce development taken by North Coast Area Health Service (NCAHS) in NSW, Australia. The NCAHS plan considers the impact of workforce ageing, changes in casemix and volume predicted by population demographics, staff level guidelines associated with service enhancements, and changes in service delivery models driven by the need for economic efficiencies, greater investment in disease prevention and medical technological advancements. Finally, the paper addresses how workforce development plans can assist sustainable service delivery through targeted strategies in recruitment, retention and retraining. PMID- 17402912 TI - A two-step multiple comparison procedure for a large number of tests and multiple treatments. AB - For situations where the number of tested hypotheses is increasingly large, the power to detect statistically significant multiple treatment effects decreases. As is the case with microarray technology, often researchers are interested in identifying differentially expressed genes for more than two types of cells or treatments. A two-step procedure is proposed for the purpose of increasing power to detect significant effects (i.e., to identify differentially expressed genes). Specifically, in the first step, the null hypothesis of equality across the mean expression levels for all treatments is tested for each gene. In the second step, only pairwise comparisons corresponding to the genes for which the treatment means are statistically different in the first step are tested. We propose an approach to estimate the overall FDR for both fixed rejection regions and fixed FDR significance levels. Also proposed is a procedure to find the FDR significance levels used in the first step and the second step such that the overall FDR can be controlled below a pre-specified FDR significance level. When compared via simulation the two-step approach has increased power over a one-step procedure, and controls the FDR at a desire significance level. PMID- 17402913 TI - Validation in genomics: CpG island methylation revisited. AB - In a recent article in PLoS Genetics, Bock et al., (2006) undertake an extensive computational epigenetics analysis of the ability of DNA sequence-derived features, capturing attributes such as tetramer frequencies, repeats and predicted structure, to predict the methylation status of CpG islands. Their suite of analyses appears highly rigorous with regard to accompanying validation procedures, employing stringent Bonferroni corrections, stratified cross validation, and follow-up experimental verification. Here, however, we showcase concerns with the validation steps, in part ascribable to the genome scale of the investigation, that serve as a cautionary note and indicate the heightened need for careful selection of analytic and companion validation methods. A series of new analyses of the same CpG island methylation data helps illustrate these issues, not just for this particular study, but also analogous investigations involving high-dimensional predictors with complex between-feature dependencies. PMID- 17402914 TI - An improved nonparametric approach for detecting differentially expressed genes with replicated microarray data. AB - Previous nonparametric statistical methods on constructing the test and null statistics require having at least 4 arrays under each condition. In this paper, we provide an improved method of constructing the test and null statistics which only requires 2 arrays under one condition if the number of arrays under the other condition is at least 3. The conventional testing method defines the rejection region by controlling the probability of Type I error. In this paper, we propose to determine the critical values (or the cut-off points) of the rejection region by directly controlling the false discovery rate. Simulations were carried out to compare the performance of our proposed method with several existing methods. Finally, our proposed method is applied to the rat data of Pan et al. (2003). It is seen from both simulations and the rat data that our method has lower false discovery rates than those from the significance analysis of microarray (SAM) method of Tusher et al. (2001) and the mixture model method (MMM) of Pan et al. (2003). PMID- 17402915 TI - Accounting for dependence in similarity data from DNA fingerprinting. AB - Differentiating strains of a pathogen is often central to investigating its epidemiological aspects. The genetic similarity of a group of strains can be assessed by calculating a matrix of dissimilarities from their DNA fingerprinting profiles. The mean dissimilarity for each strain across other strains within the group is then used as an observation in a statistical analysis. These observations are not independent of each other, and so standard analysis techniques such as the t-test are inappropriate, because they underestimate the variance of the group means, and hence overstate the statistical significance of any differences. By examining the correlation between elements of the dissimilarity matrix, it is shown that the variance is underestimated by a factor of between about 2 and 4. Permutation tests are proposed as a way of addressing the problem of dependence, and are applied to a study of fluconazole resistance in Candida albicans. PMID- 17402916 TI - Statistical inference for quantitative polymerase chain reaction using a hidden markov model: a Bayesian approach. AB - Quantitative Polymerase Chain Reaction (Q-PCR) aims at determining the initial quantity of specific nucleic acids from the observation of the number of amplified DNA molecules. The most widely used technology to monitor the number of DNA molecules as they replicate is based on fluorescence chemistry. Considering this measurement technique, the observation of DNA amplification by PCR contains intrinsically two kinds of variability. On the one hand, the number of replicated DNA molecules is random, and on the other hand, the measurement of the fluorescence emitted by the DNA molecules is collected with some random error. Relying on a stochastic model of these two types of variability, we aim at providing estimators of the parameters arising in the proposed model, and, more specifically, of the initial amount of molecules. The theory of branching processes is classically used to model the evolution of the number of DNA molecules at each replication cycle. The model is a binary splitting Galton Watson branching process. Its unknown parameters are the initial number of DNA molecules and the reaction efficiency of PCR, which is defined as the probability of replication of a DNA molecule. The number of DNA molecules is indirectly observed through noisy fluorescence measurements resulting in a so-called Hidden Markov Model. We aim at inference of the parameters of the underlying branching process, and the parameters of the noise from the fluorescence measurements in a Bayesian framework. Using simulations and experimental data, we investigate the performance of the Bayesian estimators obtained by Markov Chain Monte Carlo methods. PMID- 17402917 TI - Normalization of dye bias in microarray data using the mixture of splines model. AB - This paper discusses characteristics of dye biases in microarray data that the conventional normalization methods do not handle, and proposes a new normalization method involving a mixture of splines model. We also develop a test for between-group comparisons of each gene that is designed to be used with our proposed method. PMID- 17402918 TI - A generalized Sidak-Holm procedure and control of generalized error rates under independence. AB - For testing multiple null hypotheses, the classical approach to dealing with the multiplicity problem is to restrict attention to procedures that control the familywise error rate (FWER), the probability of even one false rejection. In many applications, one might be willing to tolerate more than one false rejection provided the number of such cases is controlled, thereby increasing the ability of the procedure to detect false null hypotheses. This suggests replacing control of the FWER by controlling the probability of k or more false rejections, which is called the k-FWER. In Hommel and Hoffmann (1987) and Lehmann and Romano (2005a), single step and stepdown procedures are derived that control the k-FWER, without making any assumptions concerning the dependence structure of the p values of the individual tests. However, if the p-values are mutually independent, one can improve the procedures. In fact, Sarkar (2005) provided such an improvement. However, we show other improvements are possible which appear to be generally much better, and are sometimes unimprovable. When k=1, the procedure reduces to the classical method of Sidak, and the stepdown procedure is unimprovable and strictly dominates that of Sarkar. Under a monotonicity condition, an unimprovable procedure is obtained. In the case k=2, the monotonicity condition is satisfied, and the condition can be checked numerically in general. We then develop a stepdown method that controls the false discovery proportion. Except for the case of k-FWER control with k=1, the gains are surprisingly dramatic, and theoretical and numerical evidence is given. PMID- 17402919 TI - Using duplicate genotyped data in genetic analyses: testing association and estimating error rates. AB - Although researchers use duplicate genotyped data to calculate an inconsistency rate, there is no power analysis to assess the value of the duplicate data. In this paper, we present a model in which the genotyping error rate is related to the inconsistency rate. We extend the g genotype by h phenotype chi-squared test to incorporate the duplicate genotyped data. When a subject is inconsistently genotyped (that is, has two observed genotypes), our procedure is to allocate 0.5 units to each of the two genotypes. We specify the multivariate analysis of variance (MANOVA) test comparing these extended counts. We provide freely available software for this test and also for a permutation test used on small samples. A simulation study shows that the asymptotic null distribution of the MANOVA test holds when the total number of subjects, N, is at least 300. We also document with a simulation study that the asymptotic distribution of this test under various alternative hypotheses is a satisfactory approximation to the simulated power. In all cases, the power of the MANOVA test using the duplicate genotyped data is greater than the power of the chi-squared test ignoring the duplicate data. Power increases ranged from 0.776% to 4.652% for 80% powered tests and 0.292% to 2.028% for 95% powered tests. Researchers now can compute the value of the duplicate genotyped data as part of the design of the study. PMID- 17402920 TI - Likelihood-based inference for multi-color optical mapping. AB - Multi-color optical mapping is a new technique being developed to obtain detailed physical maps (indicating relative positions of various recognition sites) of DNA molecules. We consider a study design in which the data consist of noisy observations of multiple copies of a DNA molecule marked with colors at recognition sites. The primary goal is to estimate a physical map. A secondary goal is to estimate error rates associated with the experiment, which are potentially useful for analysis and refinement of the biochemical steps in the mapping procedure. We propose statistical models for various sources of error and use maximum likelihood estimation (MLE) to construct a physical map and estimate error rates. To overcome difficulties arising in the maximization process, a latent-variable Markov chain version of the model is proposed, and the EM algorithm is used for maximization. In addition, a simulated annealing procedure is applied to maximize the profile likelihood over the discrete space of sequences of colors. We apply the methods to simulated data on the bacteriophage lambda genome. PMID- 17402921 TI - Sparse logistic regression with Lp penalty for biomarker identification. AB - In this paper, we propose a novel method for sparse logistic regression with non convex regularization Lp (p <1). Based on smooth approximation, we develop several fast algorithms for learning the classifier that is applicable to high dimensional dataset such as gene expression. To the best of our knowledge, these are the first algorithms to perform sparse logistic regression with an Lp and elastic net (Le) penalty. The regularization parameters are decided through maximizing the area under the ROC curve (AUC) of the test data. Experimental results on methylation and microarray data attest the accuracy, sparsity, and efficiency of the proposed algorithms. Biomarkers identified with our methods are compared with that in the literature. Our computational results show that Lp Logistic regression (p <1) outperforms the L1 logistic regression and SCAD SVM. Software is available upon request from the first author. PMID- 17402922 TI - Super learning: an application to the prediction of HIV-1 drug resistance. AB - Many alternative data-adaptive algorithms can be used to learn a predictor based on observed data. Examples of such learners include decision trees, neural networks, support vector regression, least angle regression, logic regression, and the Deletion/Substitution/Addition algorithm. The optimal learner for prediction will vary depending on the underlying data-generating distribution. In this article we introduce the "super learner", a prediction algorithm that applies any set of candidate learners and uses cross-validation to select between them. Theory shows that asymptotically the super learner performs essentially as well as or better than any of the candidate learners. In this article we present the theory behind the super learner, and illustrate its performance using simulations. We further apply the super learner to a data example, in which we predict the phenotypic antiretroviral susceptibility of HIV based on viral genotype. Specifically, we apply the super learner to predict susceptibility to a specific protease inhibitor, nelfinavir, using a set of database-derived non polymorphic treatment-selected mutations. PMID- 17402923 TI - Supervised detection of conserved motifs in DNA sequences with cosmo. AB - A number of computational methods have been proposed for identifying transcription factor binding sites from a set of unaligned sequences that are thought to share the motif in question. We here introduce an algorithm, called cosmo, that allows this search to be supervised by specifying a set of constraints that the position weight matrix of the unknown motif must satisfy. Such constraints may be formulated, for example, on the basis of prior knowledge about the structure of the transcription factor in question. The algorithm is based on the same two-component multinomial mixture model used by MEME, with stronger reliance, however, on the likelihood principle instead of more ad-hoc criteria like the E-value. The intensity parameter in the ZOOPS and TCM models, for instance, is estimated based on a profile-likelihood approach, and the width of the unknown motif is selected based on BIC. These changes allow cosmo to outperform MEME even in the absence of any constraints, as evidenced by 2- to 3 fold greater sensitivity in some simulation studies. Additional improvements in performance can be achieved by selecting the model type (OOPS, ZOOPS, or TCM) data-adaptively or by supplying correctly specified constraints, especially if the motif appears only as a weak signal in the data. The algorithm can data adaptively choose between working in a given constrained model or in the completely unconstrained model, guarding against the risk of supplying mis specified constraints. Simulation studies suggest that this approach can offer 3 to 3.5 times greater sensitivity than MEME. The algorithm has been implemented in the form of a stand-alone C program as well as a web application that can be accessed at http://cosmoweb.berkeley.edu. An R package is available through Bioconductor (http://bioconductor.org). PMID- 17402924 TI - Accurate ranking of differentially expressed genes by a distribution-free shrinkage approach. AB - High-dimensional case-control analysis is encountered in many different settings in genomics. In order to rank genes accordingly, many different scores have been proposed, ranging from ad hoc modifications of the ordinary t statistic to complicated hierarchical Bayesian models. Here, we introduce the "shrinkage t" statistic that is based on a novel and model-free shrinkage estimate of the variance vector across genes. This is derived in a quasi-empirical Bayes setting. The new rank score is fully automatic and requires no specification of parameters or distributions. It is computationally inexpensive and can be written analytically in closed form. Using a series of synthetic and three real expression data we studied the quality of gene rankings produced by the "shrinkage t" statistic. The new score consistently leads to highly accurate rankings for the complete range of investigated data sets and all considered scenarios for across-gene variance structures. PMID- 17402925 TI - The power of online role-play simulations: technology in nursing education. AB - The use and evaluation of an innovative Web-based technology and its suitability for promoting realism when interacting in clinical nursing situations are described. Also discussed are the development, implementation and evaluation of online role-play simulation. Student and faculty-centered insights are shared about the simulation and the technology supporting it. This Web-based learning experience validated literature review findings and offered new understanding about extended learning, projected realism, diversity, seriousness, non-verbal prompting, and evaluation. This simulation used the Fablusi platform in which students experienced the outcomes of their decisions and actions. This Web-based technology can be adapted for different student groups to enhance learning. PMID- 17402926 TI - When West meets East: a short-term immersion experience in South Korea. AB - The purpose of this study was to explore American student perceptions of caring for Korean patients during a 2 week exchange program. Perceptions of Korea/Koreans focused on five areas: respect, hospitality and gift giving, ability to speak English, hierarchical relationships, and being protective. Their perceptions of personal change focused in four areas: valuing personal cultural experiences, increasing cultural awareness and compassion, seeing people from other ethnic groups as individuals and developing interest in oriental medicine. Four areas of importance identified when caring for Korean patients included showing respect, importance of family, food, and care for post-partum mothers. Differences were experienced between an individualistic, low-context society (United States) and a collectivist, high context society (Korea) where the influence of Confucianism is pervasive. PMID- 17402927 TI - Teaching psychomotor skills to beginning nursing students using a web-enhanced approach: a quasi-experimental study. AB - To begin to address the problem of psychomotor skills deficiencies observed in many new graduate nurses, a skills laboratory course was developed using a web enhanced approach. In this quasi-experimental study, the control group attended weekly lectures, observed skill demonstrations by faculty, practiced skills, and were evaluated on skill performance. The experimental group learned course content using a web-enhanced approach. This allowed students to learn course material outside of class at times convenient for them, thus they had more time during class to perfect psychomotor skills. The experimental group performed better on the final cognitive examination. Students in the traditional sections were more satisfied with the course, however. It was concluded that a web enhanced approach for teaching psychomotor skills can provide a valid alternative to traditional skills laboratory formats. PMID- 17402928 TI - Comparing teaching practices about humor among nursing faculty: an international collaborative study. AB - Humor has been recognized by nurse researchers as a therapeutic intervention known to have positive psychological and physiological outcomes for patients. There is, however, no research that examines how nurses learn about humor. The purpose of this preliminary study was to examine nursing faculty members' teaching practices about humor education in the classroom and in clinical settings. Nursing faculty members from four nursing programs, two in the United States, one in Northern Ireland, and one in Taiwan, were surveyed about the inclusion of humor in the nursing curriculum. Findings revealed that substantially more humor education was included in clinical settings in the USA and Northern Ireland than in the classroom. In Taiwan, however, humor education was included more in the classroom than in clinical settings. Older and more experienced nurses with higher levels of education reported using less humor in teaching practices. PMID- 17402929 TI - Bringing theory to life: engaging nursing students in a collaborative population based screening project. AB - A collaborative population-based project for bowel cancer prevention provided an ideal opportunity to involve nursing students in applying theory to practice. In this article, described is how the engagement of students and subsequent application of a population health template contributed to a community-based bowel cancer education and screening campaign. The campaign was a valuable teaching-learning experience for students and contributed to the goal of reducing and reporting on the number of bowel cancer deaths in the local area. Project evaluation data provide insight into student learning outcomes and reveal ways to strengthen the population health initiative for future years. Originally, a scholarly pursuit of discovery and application developed in response to growing rates of bowel cancer and advances in effective screening programs, the project has evolved into the domain of teaching and learning. This evolution has benefited students, project organizers and community members. PMID- 17402930 TI - Exploring the use of clinical laboratories in undergraduate nursing programs in regional Australia. AB - Preparation for clinical practice is arguably a vital component of undergraduate nursing education with clinical laboratories widely adopted as a strategy to support student development of clinical skills. However, there is little empirical evidence about the role laboratories play in students' learning or how they assist in linking theory to practice. This study aimed to explore the current clinical laboratory practices in Schools of Nursing in regional Victoria, Australia through site visits, interviews and review of curricula. Findings revealed that approaches to laboratory learning are based on traditions rather than evidence, and have evolved in response to fiscal and environmental challenges. The predominance of teacher talk in the laboratory, has lead to acute care over other areas of practice. This study indicates a need for rigorous investigation of pedagogies that can support nursing students in preparation for clinical practice. It remains unclear if laboratory learning experiences assist students in the translation of theoretical knowledge to practice. PMID- 17402931 TI - Locating Carper's aesthetic pattern of knowing within contemporary nursing evidence, praxis and theory. AB - Carper's (1978) seminal work has been used in nursing education for many years as a method for introducing students to the multitude of ways of knowing that support nursing practice. This manuscript focuses on the aesthetic pattern of knowing and the ongoing debate in nursing literature surrounding aesthetics, evidence and nursing practice. Writers will describe and critique a strategy used to introduce first year undergraduate nursing students to this pattern of knowing. The implications of the critique of the project and the ongoing debate in relation to Carper's (1978) work, evidenced based practice and nursing praxis are discussed. PMID- 17402932 TI - Practicing what we preach: balancing teaching and clinical practice competencies. AB - Most nurse educators fulfill dual roles of clinical practitioner and teacher and thus have to achieve a balance between these two challenging sets of competencies. The authors discuss the obligation and expectation that nurse educators are concurrently experts in clinical practice and education. Is this dual competence a feasible and sustainable goal? To begin to explore this issue, the meanings of 'expert practice' and 'practice competence,' derived from the nursing education literature, are reviewed. Current professional practice competency requirements related to the nurse educator role are discussed. Questions are raised regarding support for and barriers to achieving these competencies. The potential challenges and rewards of this endeavour are presented and illustrated by two nurse educators who share their stories of achieving a balance in teaching and clinical practice competence. Finally, implications for nurse educators and directions for future research into this issue are proposed. PMID- 17402933 TI - International dimensions of higher education in nursing in Canada: tapping the wisdom of the 20th century while embracing possibilities for the 21st century. AB - New focus on the internationalization of universities occurred in the late 20th century and higher education in nursing has been quick to embrace the opportunities. In this manuscript, writers provide a brief overview of the nursing and more general literature from the late 20th century relating to key dimensions of internationalization, as well as present data from a survey conducted in 1995-96 of the international activities and dimensions at Canadian faculties/schools of nursing. While it is clear that nurses in Canadian universities were engaged in significant international endeavours in the 20th century, the literature and our experience suggest that the extent of such activity has increased substantially in recent years. Discussion centres on examination of how knowledge generated in the 20th century can inform current internationalization initiatives and on identification of key questions that merit consideration as we move forward in the 21st century. PMID- 17402934 TI - Measuring clinical practice parameters with human patient simulation: a pilot study. AB - Human Patient Simulators (HPS), electronically controlled mannequins as patient models, are increasingly being used in nursing education. However, no studies have validated the influence of systematic practice with HPS on clinical performance of nursing students. This pilot study attempted to identify the nursing clinical practice parameters influenced by HPS by evaluating the clinical performance of 12 senior second degree BSN students in five categories: safety, basic assessment skills, prioritization, problem-focused assessment, ensuing interventions, delegation and communication in a complex two-patient, simulated assignment. Students who practiced with the HPS in addition to their usual clinical training had significantly higher scores than the control group (usual clinical training alone) on Patient Identification (a subcategory of the safety category; p = 0.001), and on Assessing Vital Signs (a subcategory of the basic assessment category; p = 0.009). The control and intervention groups' performances were similar in every other category. Replication of this pilot with a larger sample is recommended. PMID- 17402935 TI - Nursing graduates' perceptions of their undergraduate clinical placement. AB - Student clinical practice is a significant and essential part of nursing education. Accordingly, clinical placements need to be positive and valuable. The purpose of this study was to describe newly-graduated nurses' perceptions of their student clinical intersession placements and how these placements impacted their functioning as graduate nurses. Inductive semantic analysis of the data revealed four themes: developing nursing skills and knowledge; preparing for future employment; experiencing supportive relationships; and experiencing realities of work-life. Educators must ensure that clinical placements are in an environment that is conducive to learning and promotes the personal and professional development of nursing students. Positive clinical experiences are more likely related to how valued and supported students feel than the physical aspects of a placement; thus, it is imperative educators assess and focus on providing placements that offer a supportive learning environment. PMID- 17402936 TI - Fostering future nursing professionals: it's a matter of values. PMID- 17402937 TI - The role and characterization of phospholipase A1 in mediating lysophosphatidylcholine synthesis in Trypanosoma brucei. AB - Lysophospholipids are ubiquitous intermediates in a variety of metabolic and signalling pathways in eukaryotic cells. We have reported recently that lysoglycerophosphatidylcholine (lyso-GPCho) synthesis in the insect form of the ancient eukaryote Trypanosoma brucei is mediated by a novel phospholipase A1 (TbPLA1). In the present study, we show that despite equal levels of TbPLA1 gene expression in wild-type insect and bloodstream trypomastigotes, both TbPLA1 enzyme levels and lysoGPCho metabolites are approx. 3-fold higher in the bloodstream form. Both of these parasite stages synthesize identical molecular species of lysoGPCho. TbPLA1 null mutants in the bloodstream form of the parasite are viable, but are deficient in lysoGPCho synthesis, a defect that can be overcome by the expression of an ectopic copy of TbPLA1. The biochemical attributes of TbPLA1-mediated lysoGPCho synthesis were examined in vitro using recombinant TbPLA1. Although TbPLA1 possesses an active-site serine residue, it is insensitive to serine-modifying reagents, such as di-isopropyl fluorophosphate and PMSF, a characteristic shared by lipases that possess lid-sheltered catalytic triads. TbPLA1 does not require metal co-factors for activity, but it does require interfacial activation prior to catalysis. Results from size-exclusion chromatography and binding kinetics analysis revealed that TbPLA1 activation by Triton X-100/GPCho mixed micelle surfaces was not specific and did not require the pre-formation of a specific enzyme-substrate complex to achieve surface binding. PMID- 17402938 TI - A novel aminopeptidase in the fat body of the moth Achaea janata as a receptor for Bacillus thuringiensis Cry toxins and its comparison with midgut aminopeptidase. AB - Bacillus thuringiensis insecticidal crystal proteins bind to cell-surface receptors which represent a family of aminopeptidases [APN (aminopeptidase N)] present on the brush border membrane of insect midgut cells of susceptible insects leading to pore formation and death of the insect. We report here for the first time the presence of a novel APN in the fat body of the moth Achaea janata. Northern blotting detected at least one APN-specific transcript in the fat body, whereas two transcripts of different sizes were detected in the midgut. We have cloned two full-length APN cDNAs of 3015 bp and 2850 bp from fat body and midgut respectively, which encode proteins of 1004 and 950 amino acids. These two APNs share only 33% amino acid sequence identity, but both display the typical APN features, such as the N-terminal signal peptide, several putative glycosylation sites, C-terminal glycosylphosphatidylinositol anchor signal, the APN-specific zinc-binding/gluzincin motif HEXXHX(18)E and gluzincin motif GAMENWG. The fat body APN manifested a variation in its expression with respect to tissue and developmental stage. In spite of the abundance of the APN transcript in the fat body, fairly low APN activity was detected in this tissue. The fat-body- and midgut-specific APNs showed differential interaction with various Cry1A toxins. Besides, the level of toxicity of different Cry subtypes varied enormously with mode/site of delivery, such as intrahaemocoelic injections and feeding bioassays. These data indicate that the fat body might be a potential alternative Cry toxin target site in the moth. PMID- 17402939 TI - Activation of biliverdin-IXalpha reductase by inorganic phosphate and related anions. AB - The effect of pH on the initial-rate kinetic behaviour of BVR-A (biliverdin IXalpha reductase) exhibits an alkaline optimum with NADPH as cofactor, but a neutral optimum with NADH as cofactor. This has been described as dual cofactor and dual pH dependent behaviour; however, no mechanism has been described to explain this phenomenon. We present evidence that the apparent peak of activity observed at neutral pH with phosphate buffer and NADH as cofactor is an anion dependent activation, where inorganic phosphate apparently mimics the role played by the 2'-phosphate of NADPH in stabilizing the interaction between NADH and the enzyme. The enzymes from mouse, rat and human all exhibit this behaviour. This behaviour is not seen with BVR-A from Xenopus tropicalis or the ancient cyanobacterial enzyme from Synechocystis PCC 6803, which, in addition to being refractory to activation by inorganic phosphate, are also differentiated by an acid pH optimum with both nicotinamide nucleotides. PMID- 17402940 TI - Editorial: pilot studies: to publish or not? PMID- 17402942 TI - Exploring the contribution of the Clinical Librarian to facilitating evidence based nursing. AB - AIM: To examine the potential role of the Clinical Librarian in facilitating evidence-based practice of nurses in acute hospital settings and develop a model for the role. BACKGROUND: There is a growing policy and professional expectation that nurses will seek out and apply evidence in their clinical practice. Studies have demonstrated that nurses experience barriers in working with an evidence based approach. The role of Clinical Librarian has been used in other countries and within medicine to overcome some of the barriers to evidence-based practice. There are limitations in the previous work in terms of rigour of evaluation, scope of the Clinical Librarian role and application to nursing in a UK setting. DESIGN: A qualitative consultation of 72 nurses in acute care settings. METHODS: Six consultation group interviews of between 4-19 participants. Written records were recorded by the scribe. Content analysis was undertaken to identify the range and frequency of comments. RESULTS: Clinical questions currently go unanswered because of barriers of time, skills deficits and access to resources. Literature searching, skills training and evidence dissemination were the main areas of work the staff requested that a Clinical Librarian should undertake. It was anticipated that the Clinical Librarian could interact and work productively with nursing staff with a limited but regular presence on the ward. Interim communication could be via e-mail, phone and written suggestions and requests for work. It was seen to be vital that the Clinical Librarian worked in partnership with staff to build evidence-based practice capacity and ensure clinical relevance of the work. CONCLUSIONS: This study has generated the first model for the Clinical Librarian role with an emphasis on nursing. It is derived from the views of clinical nurses. Recommendations are made for the implementation and evaluation of such a role. RELEVANCE TO CLINICAL PRACTICE: The Clinical Librarian could be an invaluable support to promoting evidence-based nursing. PMID- 17402943 TI - Patient participation in nursing care: towards a concept clarification from a nurse perspective. AB - AIM: The aim of this study was to investigate the meanings of the concept of patient participation in nursing care from a nurse perspective. BACKGROUND: Participation is essential and increases patients' motivation and satisfaction with received care. Studies of patient participation in nursing care are not congruent regarding definition, elements and processes. This lack of clarity is amplified by several terms used; patient/client/consumer involvement or collaboration, partnership and influence. Despite the fact that several nursing theories have emphasized the importance of patient participation, an empirically grounded theory has yet to be published. METHODS: Seven focus group interviews were held with nurses providing inpatient physical care at five hospitals in West Sweden. The focus groups consisted of Registered Swedish nurses (n = 31) who described the meaning and implementation of patient participation in nursing care. A Grounded Theory approach has been applied to tape-recorded data. Constant comparative analysis was used and saturation was achieved. RESULTS: Mutuality in negotiation emerged as the core category for explaining nurses' perspectives on patient participation in nursing care. It is characterized by four interrelated sub-core categories: interpersonal procedure, therapeutic approach, focus on resources and opportunities for influence. Mutuality in negotiation constitutes the dynamic nurse-patient interaction process. CONCLUSIONS: The study clarifies that patient participation can be explained as an interactional process identified as mutuality in negotiation based on four components. RELEVANCE TO CLINICAL PRACTICE: The results are important and can be used in nursing practice and education. Application in a clinical context means nursing care organized to include all the components presented. The results can also be used in quality assurance to improve and evaluate patient participation. PMID- 17402944 TI - Nurses' approaches towards sexuality of adolescent patients in Turkey. AB - AIMS AND OBJECTIVES: The purpose of this study is to determine the approaches regarding sexuality of nurses caring for adolescent patients in Turkey. METHOD: The sample includes nurses caring for adolescent patients in hospitals located in major cities of Turkey. There were 571 responses from the 783 nurses who were given the questionnaire. The questionnaire consists of three sections. The instrument used in this study was developed by Wall-Haas. Multiple analyses of variance in SPSS version 11.5 was used in the comparison of the scores obtained from the instrument. RESULTS: Approximately 69% of nurses stated that concerns about sexuality should be a routine component of nursing care; 70.5% of nurses stated that nursing interventions dealing with concerns about sexuality increases the patient's well being. About 70% stated that discussing sexual issues with children is the responsibility of their parents. Half of the nurses had difficulty in initiating a discussion about sexuality with their patients. CONCLUSION: The findings in this study point out that the nurses' approaches towards adolescent sexuality are rather conservative. They were not comfortable talking about issues of sexuality with adolescent patients. The nurses were aware of the importance of sexuality for an adolescent patient; however, they were not able to reflect this awareness in their practice. RELEVANCE TO CLINICAL PRACTICE: The aim of this study is to draw the nurses' attention to adolescent sexuality and to review their attitudes towards sexuality. Hence, the nurses will be able to notice their difficulties related to sexual issues and cope with them. As a result, they will be able to give holistic care to their patients. PMID- 17402945 TI - The Turkish version of the Newcastle Satisfaction with Nursing Care Scale used on medical and surgical patients. AB - AIM AND OBJECTIVES: The aim of this study is to test the validity and reliability of the modified version of the Newcastle Satisfaction with Nursing Care Scale on medical and surgical patients. BACKGROUND: Measuring patient satisfaction with nursing care is important in evaluating the extent to which patients' needs are met and for determining the appropriate nursing care. In recent years there has been increasing interest in patient satisfaction with nursing care in Turkey, but there are no validated scales available to measure this. DESIGN: It is an evaluative study. METHODS: The data were collected using the Newcastle Satisfaction with Nursing Care Scale and by a demographic information questionnaire. After translinguistic study, the content validity of the scale was confirmed and tested on 200 patients who were recruited at Istanbul University Hospital on the day of discharge. Internal consistency of the scale was tested by Cronbach's alpha. Demographic variables related to the satisfaction scores were analysed using the Spearmen correlation, the Mann-Whitney U- and Kruskal-Wallis tests. RESULTS: The Turkish version of the Satisfaction with Nursing Care Scale, with a total of 19 items, was determined to be suitable for measuring patient satisfaction with nursing care. Patients were generally satisfied with the nursing care received. The items with the most positive rating were respectively: the amount of freedom they were given on the ward, the amount of privacy they were given by nurses and how quickly nurses responded to their requests. The study found that female patients, older patients and those who had health insurance were the most satisfied. CONCLUSION: The Turkish version of the Satisfaction with Nursing Care Scale showed an adequate reliability and validity for its use on adult Turkish patients. RELEVANCE TO CLINICAL PRACTICE: Nurses can use the Satisfaction with Nursing Care Scale of Newcastle Satisfaction with Nursing Scales in evaluating and improving the nursing care in clinical practice. PMID- 17402946 TI - Early postoperative 30 degrees lateral positioning after coronary artery surgery: influence on cardiac output. AB - AIMS AND OBJECTIVES: We investigated whether: (i) Early postoperative lateral position after coronary artery bypass surgery may have a negative influence on the cardiac output and (ii) Whether turning procedures cause practical problems. BACKGROUND: Directly following surgery, coronary artery bypass patients are not receiving routine turning every two hours to prevent pressure ulcers, because a negative influence on hemodynamic parameters is assumed. DESIGN: Clinical trial. METHODS: Fifty-five coronary artery bypass patients were randomly assigned to four intervention regimens and underwent a two-hour period of 30 degrees lateral position. Fourteen patients in supine position served as a reference group. We hypothesized that 30 degrees lateral position does not cause a relevant change in the cardiac output. RESULTS: Turning the patients did not have any significant influence on the cardiac index, not even in the patients in a poor hemodynamic condition. The cardiac index in 30 degrees lateral position and supine position two to eight hours postoperatively after coronary artery bypass surgery is statistically bioequivalent. No clinically relevant deviations from preset 'safe' values for mean arterial pressure, right atrial pressure, pulmonary artery wedge pressure and pulmonary arterial pressure were observed, which would require ending the lateral position. There were no practical problems hindering the turning regimen, not even in the patients with an intra-aortic balloon pump. CONCLUSIONS: Early postoperative turning of coronary artery bypass surgery patients in lateral position is an easy and feasible procedure that does not influence the cardiac index not even in patients receiving antihypertensive or inotropic/vasopressor therapy. Further research is needed to find out whether our findings are also valid in other patient groups and other position conditions. RELEVANCE TO CLINICAL PRACTICE: If there are no strict contra-indications, lateral position has to be considered to prevent complications of continuous supine position within two hours after coronary artery bypass surgery patients have been admitted to the intensive care unit. PMID- 17402947 TI - Information needs before hospital discharge of myocardial infarction patients: a comparative, descriptive study. AB - AIMS AND OBJECTIVES: To explore the information needs of patients who have received treatment for a myocardial infarction before their discharge home from an acute hospital. WHAT IS KNOWN ABOUT THE TOPIC: Providing information for myocardial infarction patients is an important nursing function and is part of the role of health-care professionals delivering cardiac rehabilitation. It is essential to acknowledge and incorporate the self-perceived needs of patients into the information they receive. Hospital stays are becoming shorter, reducing the opportunities for nurses to provide predischarge information to patients. This highlights the challenge of adequately assessing and meeting patients' information needs. DESIGN: A comparative, descriptive survey. METHODS: A Patient Learning Needs Scale questionnaire was completed by 20 myocardial infarction patients within 72 hours of their intended discharge. Quantitative descriptive and inferential analyses were conducted using Statistical Package for Social Sciences. RESULTS: Patients indicated how important it was to know about each of 40 information items before discharge from hospital. Items related to medications, complications and physical activities were rated highly. Responses to an open question revealed that driving, returning to work and sources of support were issues of concern. Non-parametric Mann-Whitney U-tests showed that retired and older patients desired more information than their employed and younger counterparts, especially concerning community support. WHAT THE STUDY ADDS TO THE TOPIC: Previous research shows little examination of age and employment status in relation to the information needs of myocardial infarction patients. This study suggests that older and retired people may want more information than younger and employed patients. RELEVANCE TO CLINICAL PRACTICE: Older people are under represented in postdischarge cardiac rehabilitation programmes. Since these patients may need different information when discharged from younger individuals, nurses must decide how they can best meet their needs for information and continuing support. PMID- 17402948 TI - Hostility and serum homocysteine as cardiovascular risk factors in Korean patients with coronary artery disease. AB - AIMS: The aim of this study is to determine the relationships between hostility and serum homocysteine in Korean patients with coronary artery disease and to identify their effects on the occurrence of coronary artery disease. BACKGROUND: Recently, hostility as a psychosocial factor and serum homocysteine as a biochemical risk factor are gradually accepted as independent risk factors of coronary artery disease but research on the relationship between hostility and homocysteine is rare. DESIGN: This is a descriptive and correlative study conducted in the single teaching hospital located in South Korea. METHODS: Eighty four patients with coronary artery disease participated. Semi-structured interviews were used to measure the level of hostility and the characteristics of participants. Hostility was measured by the Cynical Hostility Scale and fasting blood samples from venous vein was used to measure serum homocysteine level. RESULTS: Mean scores for hostility of men were higher than women. The difference was statistically significant (p = 0.001). The serum homocysteine level (11.51 SD 5.43 micromol/l) in patients with coronary artery disease was higher than the normal reference value. Findings of this study indicate that serum homocysteine increased in a statistically consistent pattern with the level of hostility (F = 8.37, p < 0.01). CONCLUSIONS: This result suggests that high level of hostility may increase cardiovascular risk by elevated serum homocysteine concentration. RELEVANCE TO CLINICAL PRACTICE: The development and applications of nursing interventions for hostility can contribute to reducing the prevalence and mortality rate of coronary artery disease. PMID- 17402949 TI - Self-care agency and factors related to this agency among patients with hypertension. AB - AIMS AND OBJECTIVES: The aim of this study is to assess the self-care agency and factors regarding this agency among patients with hypertensions. DESIGN: This study, descriptive, analytical and cross-sectional in nature, was conducted to evaluate the self-care agency of 120 hypertensive patients at the Cardiology and Internal Diseases Polyclinic of University of Ege, Faculty of Medicine; izmir Ataturk State Hospital; University of Selcuk, Faculty of Medicine; and Karaman State Hospital between July and August 2003. METHODS: The data were collected by using the self-care agency scale and a questionnaire designed to determine the socio demographic features, and evaluated by means of percentage calculation and chi-square tests. RESULTS: The mean self-care agency rate of the patients in general was moderate. It was determined that educational situation and social insurance affected self-care. CONCLUSION: Self-care agency and health are related to economic, social and educational factors. RELEVANCE TO CLINICAL PRACTICE: Data obtained could be used for policy formulation, health planning, outcome evaluation of instruments, better health education strategies and, ultimately, to demonstrate the worth of nursing in the marketplace. PMID- 17402950 TI - Rehabilitation and guidance as reported by women and men who had undergone coronary bypass surgery. AB - AIMS AND OBJECTIVES: The purpose of this study was to describe the experience of rehabilitation reported by coronary artery disease patients who had undergone coronary artery bypass surgery. A further purpose was to describe the kind of guidance on rehabilitation that they were given during this period. METHODS: The data consisted of thematic questionnaires completed by six women and eight men who had undergone coronary artery bypass surgery. They had recorded their experiences on these questionnaires at one-month intervals for a year after the surgery. The data were analysed using the method of content analysis. RESULTS: On the basis of the results, women had experienced the most balanced period of physical, mental and social rehabilitation during the six months following surgery. Their recovery, however, suffered a setback after the first six months. On the basis of the results, men attained a better balance of physical, mental and social rehabilitation only after the six-month period following surgery. Even men had various problems in their rehabilitation, but their recovery proceeded more smoothly than that of women. Women hoped for more guidance throughout the process of rehabilitation. The support and help from wives had a positive impact on the process of men's recovery. Peer support was another significant source support. CONCLUSIONS: A person undergoing rehabilitation needs special guidance from health care professionals, especially when his/her rehabilitation does not proceed optimally. The findings suggest that, especially, the guidance of women should be improved because women seemed to suffer from many problems during their process of rehabilitation, including loneliness, insecurity, uncertainty, fears, depression and anxiety. RELEVANCE TO CLINICAL PRACTICE: Expanding the role of practical nurses to provide both preventive and rehabilitation guidance should be seen as an important strategy in health care. Individual needs, gender differences and support from families and peers should be taken into consideration when planning and implementing guidance. PMID- 17402951 TI - An examination of the intentional and unintentional aspects of medication non adherence in patients diagnosed with hypertension. AB - AIMS: The primary aim of this study was to describe the unintentional and intentional aspects of non-adherence in patients diagnosed with hypertension. A secondary aim was to examine the relationships between medication adherence and purposeful actions (intentional non-adherence), patterned behaviours (unintentional non-adherence) and demographic questionnaire variables. BACKGROUND: Non-adherence to medications continues to be a significant health care issue, the extent and consequences of which have been well documented. Despite considerable research over the past five decades, little progress has been made in solving this healthcare problem. Recent literature indicates that this lack of progress can be attributed to the fact that past research has concentrated solely upon either the unintentional or the intentional aspects of non-adherence, instead of addressing both facets simultaneously. METHODS: A quantitative, descriptive, correlation research design was employed using Johnson's (2002) Medication Adherence Model as a theoretical framework. A convenience sample of 73 participants with hypertension, attending the outpatients' clinics of two university hospitals was recruited. Data were collected by means of a researcher administered questionnaire and analysed using the Statistical Package for Social Sciences. RESULTS: High levels of medication adherence with a mean adherence score of 4.75 (maximum 5) were reported. Low and medium levels of purposeful actions and medium and high levels of patterned behaviours towards medication taking were found. Correlational analyses did not demonstrate statistically significant associations. CONCLUSIONS: Both the intentional and unintentional dimensions of medication-taking are simultaneously considered by patients to varying levels when adhering to therapeutic regimens. This is an important research area for nurses as it facilitates an increased understanding of non-adherence and, in so doing, aids the uncovering of more effective interventions aimed at sustaining lifelong pharmacotherapy. RELEVANCE TO CLINICAL PRACTICE: By acknowledging a broader approach to patient medication taking, nurses will be able more effectively to assess and intervene in non adherent behaviours and actions. PMID- 17402952 TI - Breast cancer awareness and practice of breast self examination among primary health care nurses: influencing factors and effects of an in-service education. AB - AIM: The purposes of this study were to identify the level of breast cancer and breast self examination practice knowledge of primary health nurses and the factors influencing breast cancer and breast self examination knowledge as well as to investigate the frequency of breast self examination practice of primary health nurses and influencing factors and to evaluate the effects of an in service training program about breast cancer and breast self examination practice for primary health nurses. BACKGROUND: An increased awareness among nurses about breast cancer prevention would most likely result in a higher number of better educated women about breast cancer and also motivate adherence to screening recommendations. DESIGN AND METHODS: A pre-test and post-test design was implemented to evaluate the effectiveness of a Breast Cancer and Breast Self Examination Education Program for primary health nurses. The program was instructed to 192 of the 215 primary health nurses by two nurse trainers. Pre- and post-tests made available a quantitative evaluation of the level of improvement in breast cancer and breast self-examination knowledge. RESULTS: Mean of total breast cancer knowledge score of primary health nurses prior to the program was 58.51 SD 15.63 whereas, it increased to 75.96 SD 9.53, revealing a statistically significant escalation (p < 0.0001). A significant increase was detected in the number of primary health nurses practising breast self examination as a result of the significant improvement in the mean of self breast self examination practice knowledge score which rose evidently from 71.09 SD 19.31 to 85.02 SD 12.92 (p < 0.0001). CONCLUSION: The in-service education program improved the knowledge about breast cancer and practice of breast self examination in trained primary health nurses. The program model, which involved trained primary health nurse also represented an efficient use of the resources. RELEVANCE TO CLINICAL PRACTICE: It is important that primary health nurses have adequate breast cancer knowledge and practice of breast self examination to contribute effectively to primary health care. Findings of this study can be used to guide the planning efforts on continuous education of primary health nurses about breast cancer and breast self examination issues. PMID- 17402953 TI - Working with the team: strategies employed by hospital cancer nurse specialists to implement their role. AB - AIM: This paper describes the strategies used by cancer nurse specialists in the UK to implement their role within the multiprofessional team. BACKGROUND: The prevalence of cancer increasingly preoccupies the developed world causing concern about the effective use of healthcare resources. The demand to contain healthcare costs and meet the complex needs of patients has led to the development of new and different types of nurse specialist roles. In the UK, these initiatives have been supplemented by reorganizing cancer services to ensure care is delivered by collaborative multiprofessional teams: nurse specialists are considered core team members. While role ambiguity and conflict are acknowledged as barriers to the successful introduction of new roles, little is known about the strategies used by individuals to facilitate role implementation. DESIGN: A grounded theory design using purposive and theoretical sampling. METHODS: Twenty-nine cancer nurse specialists from five hospitals participated in observation and semistructured interviews. The data were analysed concurrently using the constant comparative method. RESULTS: Acceptance, especially by doctors, was the main problem facing cancer nurse specialists. In addition, they experienced insufficient organizational support for their role. Difficulties with acceptance impaired nurses' ability to provide supportive care to cancer patients. Nurse specialists responded by employing several strategies including building relationships and establishing role boundaries. CONCLUSIONS: Some strategies used by nurse specialists are more successful than others in facilitating role implementation. While recommendations exist to assist the introduction of new roles in practice, their implementation by healthcare organizations may be limited. Future approaches should focus on helping nurses develop awareness of the problems they face, why they arise and effective mechanisms for their resolution. RELEVANCE TO CLINICAL PRACTICE: The findings highlight the mismatch between cancer policy aspirations and reality and the actions taken by nurse specialists to overcome the problems they encounter. PMID- 17402954 TI - Patient participation in decision making: views of health professionals caring for people with colorectal cancer. AB - AIM: The aim of this study was to explore views on patient participation in decision making, as described by health professionals caring for people with colorectal cancer. BACKGROUND: Patient participation in health-care decision making is on the policy agenda at an international level. However, many aspects of cancer care and treatment are complex and it is unclear how health professionals view their role as promoters of patient participation. DESIGN: A qualitative exploratory study. METHODS: In depth interviews with 35 health professionals in clinical practice. Data were analysed using thematic content analysis, assisted by a computer software package for analysis of qualitative data (N-VIVO). RESULTS: Choices in relation to surgical treatment were viewed as limited. Although it was perceived that patients could be more involved in decisions related to adjuvant treatment, providing information on various chemotherapy regimes was challenging. It was acknowledged that patients could be involved in treatment choices but there was far less clarity concerning aspects of physical and psychological care. Age was a factor when determining which patients should be offered treatment choices. CONCLUSION: The availability and presentation of choices to patients is context specific and tailored to the preferences of individuals. If health professionals focus only on aspects of decision making related to treatment, the potential for shared partnerships with patients in relation to choices about physical and psychological care may be lost. This may be particularly pertinent for nurses and allied professions who engage with patients throughout the illness trajectory. RELEVANCE TO CLINICAL PRACTICE: Policy makers should arguably appreciate that health professionals have an awareness of current thinking on patient participation, but may find policy recommendations challenging to implement in clinical practice when faced with the individual needs and preferences of patients and the complexities and uncertainties of disease management. PMID- 17402955 TI - Watching, waiting and uncertainty in prostate cancer. AB - AIM: This study explores the problems and uncertainties of older men, with prostate cancer, who have undergone watchful waiting and the strategies they use to manage their concerns. BACKGROUND: With life expectancy rising, the number of older men with prostate cancer has increased. Nearly 50% of all new prostate cancers are diagnosed in men over 70 years old. For many of these older men, improvements in medical care have increased the likelihood of living for years with prostate cancer. Thus, watchful waiting has been proposed as a reasonable alternative to treatment for older men with localized prostate cancer. METHODS: A qualitative, descriptive design was used for this study. The results from interviews with 10 men who were undergoing watchful waiting for prostate cancer were analysed using the Mishel's Reconceptualized Uncertainty in Illness model. RESULTS: Domains of uncertainty, appraisal of danger and appraisal of opportunity were identified and each was supported by participant's experience. CONCLUSIONS: The results of the study may be used to expand the Uncertainty in Illness model and develop interventions for men undergoing watchful waiting for prostate cancer. RELEVANCE TO CLINICAL PRACTICE: These findings may be used to enhance current nursing interventions for men electing for watchful waiting as an alternative to treatment for prostate cancer, including patient teaching and support services. Future study should examine the role of nurses in managing uncertainty, especially in patient teaching and in the development, implementation and evaluation of interventions to manage uncertainty. PMID- 17402956 TI - Living with dying: a hermeneutic phenomenological study of the work of hospice nurses. AB - AIMS: (i) Explore the meaning of comfort care for hospice nurses. (ii) Provide an understanding of how this work is pursued in the hospice setting. (iii) Examine the means by which hospice nurses provide comfort to hospice patients. BACKGROUND: The concepts of 'comfort' and 'comfort care' have long been a subject for examination by nurse researchers. The paper provides an overview of selected, relevant literature in this area. The methods used by nurse researchers have almost always been qualitative, and have focused on the meaning of nursing care for dying patients, from both nurses' and patients' perspectives. DESIGN AND METHODS: The paper reports a hermeneutic phenomenological study of the work of 15 hospice nurses based in one hospice in the north of England. Sampling was purposive, and data were collected by means of semi-structured interviews. A reflective diary was also kept. The interpretation of data was guided by phenomenological and hermeneutic methodology. RESULTS: The nurses interviewed spoke openly about their experiences of working with hospice patients. They saw the relief of suffering through 'comfort care' as an important element of their work. The findings are presented under three thematic headings: 'Comfort and relief', 'Peace and ease' and 'Spirituality and meaning'. CONCLUSION: Hermeneutic phenomenology is an important method for uncovering the complex realities of nursing work. The nurses' perspectives on 'comfort care' they offer to patients were revealed by the data presented here, which were interpreted to offer a unique perspective on this type of nursing work. RELEVANCE TO CLINICAL PRACTICE: These findings offer insights to nurses in both hospice and other settings; they give a number of perspectives on the nature of 'comfort care' and the meanings attached to it by experienced hospice nurses'. PMID- 17402957 TI - Esthetic abilities: a way to describe abilities of expert nurses in palliative home care. AB - AIM: The aim of this paper is to focus on the abilities needed to create the caring relation in palliative home care and to find ways to describe these abilities and skills from an esthetic perspective. BACKGROUND: Nurses in Palliative home care have to create a functioning relationship between themselves the patient and the patient's next of kin, this puts special demands on the nurse. A number of abilities, such as creativity, intuition, empathy and self knowledge are mentioned in literature related to the caring relation. Many nursing theorists have referred to the art and esthetic of nursing when trying to describe these abilities. METHODS: Data were collected using semi-structured interviews with eight expert nurses in palliative home care. The transcribed interviews were analysed using qualitative content analyses. FINDINGS: Three main categories where found: The will to do good, Knowledge and Perceptiveness. Subcategories that can be seen as abilities where found in the main categories knowledge and perceptiveness. CONCLUSIONS: The main categories can be seen as expressions for abilities, personal qualities and skills needed to create the caring relation in palliative home care. We found interesting connections between the three main categories and the concepts of esthetics, ethics and science. We also found that nurses develop in a way, i.e. similar to an artist. RELEVANCE TO CLINICAL PRACTICE: The concepts brought forward in this paper could be used in clinical supervision and education as well as in clinical practice. If nurses think about the three aspects: Knowledge, the will to do good and perceptiveness, the chance for a positive relation between the nurse and the patient, and the patient's next of kin might increase. One could speculate that perceptiveness is a vital ability in order to achieve the expert level of nursing and that this perspective should be taken in consideration when recruiting nurses and in curriculum development. PMID- 17402958 TI - Attitudes towards holistic complementary and alternative medicine: a sample of healthy people in Turkey. AB - AIMS AND OBJECTIVES: This study aimed to investigate the attitude towards holistic complementary and alternative medicine of healthy people, and to evaluate the relationship between attitude towards holistic complementary and alternative medicine and the characteristics of the participants. BACKGROUND: Complementary and alternative medicines are becoming more accepted. DESIGN: This study used descriptive and correlational designs. METHODS: The study included healthy individuals who attended or visited a primary care centre for healthcare services. The sample of the study consisted of 448 persons who responded to the questionnaire. The Attitude towards Holistic Complementary and Alternative Medicine scale consisted of 11 items on a six-point, and two subscales. The mean score of holistic complementary and alternative medicine was studied in relation to attributes and holistic complementary and alternative medicine. RESULTS: The mean score on the scale was 58.1 SD 4.1 point, and in terms of the mean score of the scale, the sample group showed a negative attitude towards holistic complementary and alternative medicine and one subscale. Demographic characteristics of the sample group affected attitudes towards holistic complementary and alternative medicine and both subscales. CONCLUSION: In light of these results, it is clear that healthy Turkish population have a tendency towards conventional medicine. RELEVANCE TO CLINICAL PRACTICE: Health professionals caring for healthy people should provide comprehensive care that addresses the physical, psychosocial and spiritual needs of the individual; they could provide the consultation regarding to different patterns of complementary therapies. PMID- 17402959 TI - Qigong practice among chronically ill patients during the SARS outbreak. AB - AIMS AND OBJECTIVES: This research aimed at exploring the motivations and experiences of chronically ill participants practising qigong during the severe acute respiratory syndrome outbreak in Hong Kong. BACKGROUND: Although biomedicine is the mainstream medical system in Hong Kong, many people employ complementary and alternative medicine in dealing with their chronic health problems. Practising qigong is one of the most popular forms of complementary and alternative medicine used among chronically ill patients. Little is known about the experiences of the chronically ill patients practising qigong in the severe acute respiratory syndrome outbreak and even less is known how this practice is related to the social context of Hong Kong during the outbreak. DESIGN AND METHODS: Qualitative methods using participant-observation from three qigong classes in Hong Kong and in-depth semi-structured interviews with 30 participants were employed. Content analysis and discourse analysis were used to identify major themes of the data. RESULTS: Both the underlying and trigger motivations could motivate these chronically ill participants to practise qigong. Legitimacy of qigong in health maintenance, deterioration of health and unpleasant experiences in biomedical treatment on their chronic illnesses served as the underlying motivations. Stigmatization of and discrimination against the chronically ill and the sense of searching coping strategy during the severe acute respiratory syndrome outbreak further motivated their practice. CONCLUSION: To the participants, practising qigong not only could strengthen their health, but was also a coping strategy for them to regain an active control and the sense of security in their health and enabled them to overcome the social stigmatization and discrimination during the outbreak. RELEVANCE TO CLINICAL PRACTICE. Searching for emotional support and a more active role in health conditions could be seen from the chronically ill participants. Understanding and paying more attention to the particular needs of the chronically ill patients can enable health-care professionals to provide better care and support for the chronically ill during an epidemic crisis. PMID- 17402960 TI - Does the experimental design capture the effects of complementary therapy? A study using reflexology for patients undergoing coronary artery bypass graft surgery. AB - AIM: Our purpose was to pilot test whether reflexology may reduce anxiety in patients undergoing Coronary Artery Bypass Graft Surgery in Iceland. BACKGROUND: Nurses need to study the effects of complementary therapies in general and particularly those that may be beneficial to decrease patients' anxiety. It has been assumed that reflexology lessens anxiety, but research is needed to substantiate such expectations. DESIGN: A pilot study using randomized design with experimental and control groups. METHODS: Nine patients were recruited and randomly assigned into groups with five patients assigned into an experimental group receiving reflexology for 30 minutes and four patients into control group which rested for 30 minutes. Anxiety and physiological variables were measured pre- and post-reflexology sessions once a day over five days. RESULTS: The anxiety scores were lower for patients in the control group on all measures. Systolic blood pressure lowered significantly more in the control group than in the treatment group. No significant changes were observed for other variables. Patients' comments and responses overwhelmingly suggested increased well-being due to both experimental and control intervention. CONCLUSION: This study showed little evidence to support reflexology as a mean of reducing anxiety in CABG patients. Several methodological problems were identified that need to be considered further. RELEVANCE TO CLINICAL PRACTICE: It is suggested that reflexology should be tailored to individual needs and research methods used that allow for capturing its holistic nature. Further scholarly work is warranted to explore several methodological issues in studying complementary therapies in a highly complex treatment situation. PMID- 17402961 TI - Application of a virtual reality prototype for pain relief of pediatric burn in Taiwan. AB - AIM: This study examines the usability and effectiveness of virtual reality in reducing pain in wound-care procedures for pediatric burn patients in Taiwan. BACKGROUND: Virtual reality has continuously gained prominence in the medical arena, for instance, the telepresence for surgery, the management of mental health disorders and pain control of the paediatric burn. Notwithstanding an increased application of virtual reality in the medical arena in North America, there have been no studies investigating its use for paediatric burn patients in Asia. METHODS: This descriptive study has two phases: Phase I: the development of a virtual reality prototype. Phase II: the implementation of the prototype to discern its usability and efficacy with paediatric burn patients at a local hospital. RESULTS: The findings suggest that a significant difference is found in the children's reported pain, with or without the virtual reality intervention, over the three phases: before, during and after the dressing change. However, less pain was noted in the intervention group during and after the dressing change. CONCLUSION: Adding to the existing clinical value of virtual reality identifies the nature of and different children's responses to pain with the use of virtual reality. RELEVANCE TO CLINICAL PRACTICE: This study is significant since it demonstrates a difference in the child's response to pain based on the nature of presence and distraction. Moreover, given the evidence that a decrease in anxiety was experienced after the dressing change with virtual reality intervention, timing of using the virtual reality intervention before the child develops conditioning anxiety and anticipated pain for the procedure would be of importance. PMID- 17402962 TI - Self-administered acupressure reduces the symptoms that limit daily activities in bronchiectasis patients: pilot study findings. AB - AIMS AND OBJECTIVES: To examine and compare the effects of acupressure on the perceived health-related quality of life of the participants with bronchiectasis. BACKGROUND: In an attempt to offer comfort, pain control and symptom management, nursing is becoming increasingly involved in offering complementary-alternative medicine as part of its caring-healing focus in comprehensive patient care. Acupressure is one such modality that is being increasingly used by both medical and nursing professionals. While acupressure has been reported to have beneficial effects in patients with respiratory disease, the benefits to bronchiectasis patients have remained uncertain. DESIGN: A randomized, partially blinded study consisting of three groups. METHODS: Thirty-five out-patients of both genders, aged 59.46 SD 11.52 years, who were suffering from bronchiectasis, were randomly split into one of three groups: standard care with supplemental acupressure for eight weeks (11 participants); standard care with supplemental sham acupressure for eight weeks (11 participants); and standard care alone (13 participants). Outcomes were determined by changes in daily sputum amounts, sputum self assessment, six-minute walking distance, breathing difficulty (measured on the dyspnea visual analogue scale) and health-related quality of life (measured by the Saint George Respiratory Questionnaire). RESULTS: The sputum self-assessment score improved over time for the sham acupressure participants (P = 0.03), when compared with the controls. For acupressure participants, the Saint George respiratory questionnaire activity component scores also improved over time, compared with controls (P = 0.01) after adjustment for covariates (treatment, time, age, sex and baseline values). Other variables did not differ between the standard care alone group and the other two groups. CONCLUSIONS: Eight weeks of self-administered acupressure could be useful in reducing the effects of bronchiectasis on a patient's daily activities. RELEVANCE TO CLINICAL PRACTICE: Acupressure may be regarded as a viable nursing intervention. PMID- 17402963 TI - Commentary on Sun F-K, Long A, Boore J & Lee-Ing T (2006) Patients and nurses' perceptions of ward environmental factors and support systems in the care of suicidal patients. Journal of Clinical Nursing 15, 83-92. PMID- 17402966 TI - Role of activity-dependent mechanisms in the control of dopaminergic neuron survival. AB - Dopaminergic neurons that constitute the nigrostriatal pathway are characterized by singular electrical properties that allow them to discharge in vivo spontaneously in a spectrum of patterns ranging from pacemaker to random and bursting modes. These electrophysiological features allow dopaminergic neurons to optimize the release of dopamine in their terminal fields. However, there is emerging evidence indicating that electrical activity might also participate in the control of dopaminergic neuron survival, not only during development, but also in the adult brain, thus raising the possibility that alterations in ionic currents could contribute actively to the demise of these neurons in Parkinson disease. This review focuses on the mechanisms by which activity-dependent mechanisms might modulate dopaminergic cell survival. PMID- 17402967 TI - Alternative isoforms of myelin/oligodendrocyte glycoprotein with variable cytoplasmic domains are expressed in human brain. AB - The human myelin/oligodendrocyte glycoprotein (MOG) gene is encoded by 10 exons that exhibit a complex pattern of alternative splicing. This report demonstrates that several MOG-specific alternative splice variants are indeed expressed in human oligodendrocytes (OLs) and myelin during perinatal development and are retained through adulthood. While all forms possess the common extracellular Ig like domain, these alternative MOG structures differ significantly in their respective cytoplasmic domains. Peptide-specific antibodies were generated to facilitate detection of these different MOG moieties. The fidelity of these antibodies is shown using N20 OLs expressing individual MOG variants. These antibodies also only co-localize with another well-characterized marker of OLs and myelin--PLP/DM20 proteins. Among the human tissue samples tested, very limited expression occurred by 36 weeks gestation for 2-3 MOG variants, and the remaining MOG isoforms were not evident until shortly after birth. This study represents the first evidence of alternative translation products from the MOG gene. To date, it is believed that alternative splicing of MOG is limited to primates. Recent completion of various genome projects has revealed that alternative splicing is much more prevalent than originally estimated, and species-specific alternative splicing is now being shown to be highly relevant to expanding proteomic diversity. PMID- 17402968 TI - Metabotropic glutamate receptor 3 protects neurons from glucose-induced oxidative injury by increasing intracellular glutathione concentration. AB - High glucose concentrations cause oxidative injury and programmed cell death in neurons, and can lead to diabetic neuropathy. Activating the type 3 metabotropic glutamate receptor (mGluR3) prevents glucose-induced oxidative injury in dorsal root ganglion neurons co-cultured with Schwann cells. To determine the mechanisms of protection, studies were performed in rat dorsal root ganglion neuron-Schwann cell co-cultures. The mGluR3 agonist 2R,4R-4-aminopyrrolidine-2,4-dicarboxylate prevented glucose-induced inner mitochondrial membrane depolarization, reactive oxygen species accumulation, and programmed cell death, and increased glutathione (GSH) concentration in co-cultured neurons and Schwann cells, but not in neurons cultured without Schwann cells. Protection was diminished in neurons treated with the GSH synthesis inhibitor l-buthionine-sulfoximine, suggesting that mGluR mediated protection requires GSH synthesis. GSH precursors and the GSH precursor GSH-ethyl ester also protected neurons from glucose-induced injury, indicating that GSH synthesis in Schwann cells, and transport of reaction precursors to neurons, may underlie mGluR-mediated neuroprotection. These results support the conclusions that activating glial mGluR3 protects neurons from glucose-induced oxidative injury by increasing free radical scavenging and stabilizing mitochondrial function, through increased GSH antioxidant defense. PMID- 17402969 TI - Neuroprotective role of bradykinin because of the attenuation of pro-inflammatory cytokine release from activated microglia. AB - Bradykinin (BK) has been reported to be a mediator of brain damage in acute insults. Receptors for BK have been identified on microglia, the pathologic sensors of the brain. Here, we report that BK attenuated lipopolysaccharide (LPS) induced release of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta from microglial cells, thus acting as an anti-inflammatory mediator in the brain. This effect was mimicked by raising intracellular cAMP or stimulating the prostanoid receptors EP2 and EP4, while it was abolished by a cAMP antagonist, a prostanoid receptor antagonist, or by an inhibitor of the inducible cyclooxygenase (cyclooxygenase-2). BK also enhanced formation of prostaglandin E(2) and expression of microsomal prostaglandin E synthase. Expression of BK receptors and EP2/EP4 receptors were also enhanced. Using physiological techniques, we identified functional BK receptors not only in culture, but also in microglia from acute brain slices. BK reduced LPS-induced neuronal death in neuron-microglia co-cultures. This was probably mediated via microglia as it did not affect TNF-alpha-induced neuronal death in pure neuronal cultures. Our data imply that BK has anti-inflammatory and neuroprotective effects in the central nervous system by modulating microglial function. PMID- 17402970 TI - Group I metabotropic glutamate receptor-dependent TRPC channel trafficking in hippocampal neurons. AB - The group I metabotropic glutamate receptor agonist (S)-3,5 dihydroxyphenylglycine (DHPG) elicited two phases of synchronized neuronal (epileptiform) discharges in hippocampal slices: an initial phase of short duration discharges followed by a phase of prolonged discharges. We assessed the involvement of transient receptor potential canonical (TRPC) channels in these responses. Pre-treatment of hippocampal slices with TRPC channel blockers, 1 [beta-[3-(4-methoxyphenyl)propoxy]-4-methoxyphenethyl]-1H-imidazole hydrochloride (SKF96365) or 2-aminoethoxydiphenyl borate, did not affect the short epileptiform discharges but blocked the prolonged epileptiform discharges. SKF96365 suppressed ongoing DHPG-induced prolonged epileptiform discharges. Western blot analysis showed that the total TRPC4 or TRPC5 proteins in hippocampal slices were unchanged following DHPG. DHPG increased TRPC4 and TRPC5 in the cytoplasmic compartment and decreased these proteins in the plasma membrane. Translocation of TRPC4 and TRPC5 was suppressed when the epileptiform discharges were blocked by ionotropic glutamate receptor blockers. Translocation of TRPC4 and TRPC5 was also prevented in slices from phospholipase C (PLC) beta1 knockout mice, even when synchronized discharges were elicited by the convulsant 4-aminopyridine. The results suggest that TRPC channels are involved in generating DHPG-induced prolonged epileptiform discharges. This function of TRPC channels is associated with a neuronal activity- and PLCbeta1-dependent translocation of TRPC4 and TRPC5 proteins from the plasmalemma to the cytoplasmic compartment. PMID- 17402971 TI - Nitric oxide synthase inhibitors modulate nerve growth factor-mediated regulation of amyloid precursor protein expression in PC12 cells. AB - Nerve growth factor (NGF) can regulate nitric oxide synthase (NOS) expression and nitric oxide (NO) can modulate NGF-mediated neurotrophic responses. In this study, the role of NO in NGF-stimulated amyloid precursor protein (APP) levels was studied. PC12 cells were treated with either the non-selective NOS inhibitor N(omega)-nitro-L-arginine methylester (L-NAME) or the inducible NOS selective inhibitor s-methylisothiourea (S-MIU), and the effect on NGF-mediated increases in APP expression was determined. NGF significantly increased total APP protein levels following 96 h of treatment and this increase was prevented in cells pre treated with S-MIU. Pre-treatment of cells with actinomycin D also blocked this NGF-mediated induction of APP, indicating de novo protein synthesis is necessary. Treatment with NGF increased APP promoter activity; however, this increase was only partially inhibited by pre-treatment with S-MIU and was increased in the presence of L-NAME. This suggests that NO may be modulating other aspects of APP expression in addition to transcription. Inhibition of NGF signaling pathways was also investigated using inhibitors of mitogen-activated protein (MAP) kinase (U0126), Akt (LY294002) and protein kinase C (PKC; U73122 and bisindolylmaleimide 1 (BIS-1)) activation. Inhibition of each of these pathways prevented NGF mediated increases in APP protein expression; however, only BIS-1 attenuated NGF mediated increases in promoter activation. This study indicates that NO is involved in the NGF-mediated regulation of APP, in part at the level of APP transcription and could involve the modulation of NGF signal transduction pathways. PMID- 17402972 TI - Topological analysis of the complex formed between neurokinin A and the NK2 tachykinin receptor. AB - Neurokinin A stimulates physiological responses in the peripheral and central nervous systems upon interacting primarily with the tachykinin NK2 receptor (NK2R). In this study, the structure of NKA bound to the NK2R is characterised by use of fluorescence resonance energy transfer. Four fluorescent NKA analogues with Texas red introduced at amino acid positions 1, 4, 7 and 10 were prepared. When bound to a NK2R carrying enhanced green fluorescent protein at the N terminus, all peptides reduce green fluorescent protein fluorescence from 10% to 50% due to energy transfer. The derived donor-acceptor distances are 46, 55, 59 and 69 A for the fluorophore linked to positions 1-10, respectively. The monotonic increase in distance clearly indicates that the peptide adopts an extended structure when bound to its receptor. The present data are used, in combination with rhodopsin structure, fluorescence studies, photoaffinity labelling and site-directed mutagenesis data to design a computer model of the NKA-NK2R complex. We propose that the N-terminus of NKA is exposed and accessible to the extracellular medium. Subsequent amino acids of the NKA peptide become progressively more buried residues up to approximately one-third of the transmembrane-spanning domain. PMID- 17402973 TI - Axonal involvement in the Wlds neuroprotective effect: analysis of pure motoneurons in a mouse model protected from motor neuron disease at a pre symptomatic age. AB - The identification of the Wlds gene that delays axonal degeneration in several models of neurodegenerative disease provides an interesting tool to study mechanisms of axonal loss. We showed that crossing a mouse mutant with a motoneuron disease (pmn for progressive motor neuronopathy) with mice that express the Wlds gene delayed axonal loss, increased the life span, partially rescued axonal transport deficit and prolonged the survival of the motoneuron cell bodies. To determine factors involved in the neuroprotective effect of Wlds, we combined laser capture microdissection and microarray analysis to identify genes that are differentially regulated at a pre-symptomatic age in motoneuron cell bodies in pmn/pmn,Wlds/Wlds mice as compared with pmn/pmn mice. Only 56 genes were de-regulated; none of the 'classical' genes implicated in apoptosis were de-regulated. Interestingly, a large proportion of these genes are related to axonal function and to retrograde and anterograde transport (i.e. members of the dynactin complex and kinesin family). These results were confirmed by real time PCR, in situ hybridization and at protein level in sciatic nerves. Thus, genes related to axonal function and in particular to axonal transport may be involved at an early stage in the neuroprotective property of the Wlds gene and confirm the importance of axonal involvement in this model of motor neuron disease. PMID- 17402974 TI - Advances in our understanding of mammalian sex-biased dispersal. AB - Sex-biased dispersal is an almost ubiquitous feature of mammalian life history, but the evolutionary causes behind these patterns still require much clarification. A quarter of a century since the publication of seminal papers describing general patterns of sex-biased dispersal in both mammals and birds, we review the advances in our theoretical understanding of the evolutionary causes of sex-biased dispersal, and those in statistical genetics that enable us to test hypotheses and measure dispersal in natural populations. We use mammalian examples to illustrate patterns and proximate causes of sex-biased dispersal, because by far the most data are available and because they exhibit an enormous diversity in terms of dispersal strategy, mating and social systems. Recent studies using molecular markers have helped to confirm that sex-biased dispersal is widespread among mammals and varies widely in direction and intensity, but there is a great need to bridge the gap between genetic information, observational data and theory. A review of mammalian data indicates that the relationship between direction of sex-bias and mating system is not a simple one. The role of social systems emerges as a key factor in determining intensity and direction of dispersal bias, but there is still need for a theoretical framework that can account for the complex interactions between inbreeding avoidance, kin competition and cooperation to explain the impressive diversity of patterns. PMID- 17402975 TI - Differential admixture shapes morphological variation among invasive populations of the lizard Anolis sagrei. AB - The biological invasion of the lizard Anolis sagrei provides an opportunity to study evolutionary mechanisms that produce morphological differentiation among non-native populations. Because the A. sagrei invasion represents multiple native range source populations, differential admixture as well as random genetic drift and natural selection, could shape morphological evolution during the invasion. Mitochondrial DNA (mtDNA) analyses reveal seven distinct native-range source populations for 10 introduced A. sagrei populations from Florida, Louisiana and Texas (USA), and Grand Cayman, with 2-5 native-range sources contributing to each non-native population. These introduced populations differ significantly in frequencies of haplotypes from different native-range sources and in body size, toepad-lamella number, and body shape. Variation among introduced populations for both lamella number and body shape is explained by differential admixture of various source populations; mean morphological values of introduced populations are correlated with the relative genetic contributions from different native range source populations. The number of source populations contributing to an introduced population correlates with body size, which appears independent of the relative contributions of particular source populations. Thus, differential admixture of various native-range source populations explains morphological differences among introduced A. sagrei populations. Morphological differentiation among populations is compatible with the hypothesis of selective neutrality, although we are unable to test the hypothesis of interdemic selection among introductions from different native-range source populations. PMID- 17402976 TI - Differentiation of sympatric populations of the band-rumped storm-petrel in the Galapagos Islands: an examination of genetics, morphology, and vocalizations. AB - In each of at least two locations within the Galapagos Islands, breeding band rumped storm-petrels (Oceanodroma castro) form two distinct populations that use the same colony site at separate times of the year for reproduction. Temporal segregation of these populations raises the possibility that they are reproductively isolated and represent cryptic species. We examined variation in mitochondrial DNA, morphology, and vocalizations of storm-petrel populations nesting 6 months apart on the islet of Plaza Norte in the Galapagos. Seasonal populations displayed low but significant levels of differentiation in the mitochondrial control region, five morphological variables, and one feature of male vocalizations. Breeding populations appear to have been separated for approximately 1700 years. Given the recent divergence date and relatively high effective population sizes (4000-5600 females each), seasonal populations are unlikely to be in genetic equilibrium. As a result, the low divergence estimate probably reflects historical association and not contemporary genetic exchange. These populations are not sufficiently differentiated to be considered cryptic species. However, they are probably in the early stages of divergence. Consequently, we recommend that cool- and hot season populations on Plaza Norte be recognized as separate management units. PMID- 17402977 TI - Broad- to fine-scale population genetic patterning in the smallmouth bass Micropterus dolomieu across the Laurentian Great Lakes and beyond: an interplay of behaviour and geography. AB - Analysis of population genetic relationships reveals the signatures of current processes such as spawning behaviour and migration, as well as those of historical events including vicariance and climate change. This study examines these signatures through testing broad- to fine-scale genetic patterns among smallmouth bass Micropterus dolomieu spawning populations across their native Great Lakes range and outgroup areas, with fine-scale concentration in Lake Erie. Our primary hypotheses include whether genetic patterns result from behavioural and/or geographical isolation, specifically: (i) Are spawning groups in interconnected waterways genetically separable? (ii) What is the degree of isolation across and among lakes, basins, and tributaries? (iii) Do genetic divergences correspond to geographical distances? and (iv) Are historical colonization patterns from glacial refugia retained? Variation at eight nuclear microsatellite DNA loci are analysed for 666 smallmouth bass from 28 locations, including 425 individuals in Lake Erie; as well as Lakes Superior, Huron, and Ontario, and outgroups from the Mississippi, Ohio, St. Lawrence, and Hudson River drainages. Results reveal marked genetic differences among lake and river populations, as well as surprisingly high divergences among closely spaced riverine sites. Results do not fit an isolation-by-geographical-distance prediction for fine-scale genetic patterns, but show weak correspondence across large geographical scales. Genetic relationships thus are consistent with hypotheses regarding divergent origins through vicariance in glacial refugia, followed by colonization pathways establishing modern-day Great Lakes populations, and maintenance through behavioural site fidelity. Conservation management practices thus should preserve genetic identity and unique characters among smallmouth bass populations. PMID- 17402978 TI - The influence of altitude and topography on genetic structure in the long-toed salamander (Ambystoma macrodactulym). AB - A primary goal of molecular ecology is to understand the influence of abiotic factors on the spatial distribution of genetic variation. Features including altitudinal clines, topography and landscape characteristics affect the proportion of suitable habitat, influence dispersal patterns, and ultimately structure genetic differentiation among populations. We studied the effects of altitude and topography on genetic variation of long-toed salamanders (Ambystoma macrodactylum), a geographically widespread amphibian species throughout northwestern North America. We focused on 10 low altitude sites (< 1200 m) and 11 high-altitude sites in northwestern Montana and determined multilocus genotypes for 549 individuals using seven microsatellite loci. We tested four hypotheses: (1) gene flow is limited between high- and low-altitude sites; and, (2) gene flow is limited among high-altitude sites due to harsh habitat and extreme topographical relief between sites; (3) low-altitude sites exhibit higher among site gene flow due to frequent flooding events and low altitudinal relief; and (4) there is a negative correlation between altitude and genetic variation. Overall F(ST) values were moderate (0.08611; P < 0.001). Pairwise F(ST) estimates between high and low populations and a population graphing method supported the hypothesis that low-altitude and high-altitude sites, taken together, are genetically differentiated from each other. Also as predicted, gene flow is more prominent among low-altitude sites than high-altitude sites; low-altitude sites had a significantly lower F(ST) (0.03995; P < 0.001) than high altitude sites (F(ST) = 0.10271; P < 0.001). Use of Bayesian analysis of population structure (BAPS) resulted in delineation of 10 genetic groups, two among low-altitude populations and eight among high-altitude populations. In addition, within high altitude populations, basin-level genetic structuring was apparent. A nonequilibrium algorithm for detecting current migration rates supported these population distinctions. Finally, we also found a significant negative correlation between genetic diversity and altitude. These results are consistent with the hypothesis that topography and altitudinal gradients shape the spatial distribution of genetic variation in a species with a broad geographical range and diverse life history. Our study sheds light on which key factors limit dispersal and ultimately species' distributions. PMID- 17402979 TI - Panmictic population structure in the hooded seal (Cystophora cristata). AB - Two putative populations of hooded seals (Cystophora cristata) occur in the North Atlantic. The Greenland Sea population pup and breed on the pack ice near Jan Mayen ('West Ice') while the Northwest Atlantic population is thought to pup in the Davis Strait, in the Gulf of St. Lawrence (the 'Gulf'), and off southern Labrador or northeast Newfoundland (the 'Front'). We used microsatellite profiling of 300 individuals at 13 loci and mitochondrial DNA sequencing of the control region of 123 individuals to test for genetic differentiation between these four breeding herds. We found no significant genetic differences between breeding areas, nor evidence for cryptic nor higher level genetic structure in this species. The Greenland Sea breeding herd was genetically most distant from the Northwest Atlantic breeding areas; however, the differences were statistically nonsignificant. Our data therefore suggest that the world's hooded seals comprise a single panmictic genetic population. PMID- 17402980 TI - Population genetic consequences of geographic disjunction: a prairie plant isolated on Great Lakes alvars. AB - Species may often exhibit geographic variation in population genetic structure due to contemporary and historical variation in population size and gene flow. Here, we test the predictions that populations on the margins of a species' distribution contain less genetic variation and are more differentiated than populations towards the core of the range by comparing patterns of genetic variation at five microsatellite loci between disjunct and core populations of the perennial, allohexaploid herb Geum triflorum. We sampled nine populations isolated on alvar habitat within the eastern Great Lakes region in North America, habitats that include disjunct populations of several plant species, and compared these to 16 populations sampled from prairie habitat throughout the core of the species' distribution in midwestern Canada and the USA. Alvar populations exhibited much lower within-population diversity and contained only a subset of alleles found in prairie populations. We detected isolation by distance across the species' range and within alvar and prairie regions separately. As predicted, genetic differentiation was higher among alvar populations than among prairie populations, even after controlling for the geographic distance between sampled populations. Low diversity and high differentiation can be accounted for by the greater contemporary spatial isolation of alvar populations. However, the genetic structure of alvar populations may also have been influenced by postglacial range expansion and contraction. Our results are consistent with alvar populations being founded during an expansion of prairie habitat during the warmer, hypsithermal period approximately 5000 bp and subsequently becoming stranded on isolated alvar habitat as the climate grew cooler and wetter. PMID- 17402981 TI - From the Apennines to the Alps: colonization genetics of the naturally expanding Italian wolf (Canis lupus) population. AB - Wolves in Italy strongly declined in the past and were confined south of the Alps since the turn of the last century, reduced in the 1970s to approximately 100 individuals surviving in two fragmented subpopulations in the central-southern Apennines. The Italian wolves are presently expanding in the Apennines, and started to recolonize the western Alps in Italy, France and Switzerland about 16 years ago. In this study, we used a population genetic approach to elucidate some aspects of the wolf recolonization process. DNA extracted from 3068 tissue and scat samples collected in the Apennines (the source populations) and in the Alps (the colony), were genotyped at 12 microsatellite loci aiming to assess (i) the strength of the bottleneck and founder effects during the onset of colonization; (ii) the rates of gene flow between source and colony; and (iii) the minimum number of colonizers that are needed to explain the genetic variability observed in the colony. We identified a total of 435 distinct wolf genotypes, which showed that wolves in the Alps: (i) have significantly lower genetic diversity (heterozygosity, allelic richness, number of private alleles) than wolves in the Apennines; (ii) are genetically distinct using pairwise F(ST) values, population assignment test and Bayesian clustering; (iii) are not in genetic equilibrium (significant bottleneck test). Spatial autocorrelations are significant among samples separated up to c. 230 km, roughly correspondent to the apparent gap in permanent wolf presence between the Alps and north Apennines. The estimated number of first-generation migrants indicates that migration has been unidirectional and male-biased, from the Apennines to the Alps, and that wolves in southern Italy did not contribute to the Alpine population. These results suggest that: (i) the Alps were colonized by a few long-range migrating wolves originating in the north Apennine subpopulation; (ii) during the colonization process there has been a moderate bottleneck; and (iii) gene flow between sources and colonies was moderate (corresponding to 1.25-2.50 wolves per generation), despite high potential for dispersal. Bottleneck simulations showed that a total of c. 8-16 effective founders are needed to explain the genetic diversity observed in the Alps. Levels of genetic diversity in the expanding Alpine wolf population, and the permanence of genetic structuring, will depend on the future rates of gene flow among distinct wolf subpopulation fragments. PMID- 17402982 TI - Genetic differentiation between Principe Island and mainland populations of the grey parrot (Psittacus erithacus), and implications for conservation. AB - The range of the grey parrot (Psittacus erithacus), one of the most heavily harvested bird species for the international pet trade, spans the forest belt of Central and West Africa and includes the oceanic island of Principe (Gulf of Guinea). Morphological variation led to the recognition of two mainland subspecies (P. e. erithacus and P. e. timneh). The population from Principe was originally described as a separate species (P. princeps) but is currently included in the nominate race. We used 1932 bp of the mitochondrial genome to clarify the genealogical affinities between the two currently-recognized mainland subspecies and the Principe population. Sampling included 20 individuals from Principe, 17 from P. e. erithacus, and 13 from P. e. timneh. We found that the two mainland subspecies form two independent lineages, having diverged up to 2.4 million years ago (Ma), and that the Principe population is composed of two lineages that diverged in allopatry. The most common lineage is descended from the first colonizers and evolved in isolation for up to 1.4 Ma. Contrary to current understanding, this 'Principe lineage' is more closely related to the timneh than to the erithacus subspecies. The second lineage consists of P. e. erithacus birds from the nearby mainland that colonized the island in recent times. The evolutionary dynamics of the grey parrot population of Principe are primarily characterized by isolation, with new genetic variation being added through rare immigration events. The heavily harvested Principe grey parrot population should therefore be treated as an independent conservation unit. PMID- 17402983 TI - Lack of genetic structure among ecologically adapted populations of an Australian rainforest Drosophila species as indicated by microsatellite markers and mitochondrial DNA sequences. AB - Although fragmented rainforest environments represent hotspots for invertebrate biodiversity, few genetic studies have been conducted on rainforest invertebrates. Thus, it is not known if invertebrate species in rainforests are highly genetically fragmented, with the potential for populations to show divergent selection responses, or if there are low levels of gene flow sufficient to maintain genetic homogeneity among fragmented populations. Here we use microsatellite markers and DNA sequences from the mitochondrial ND5 locus to investigate genetic differences among Drosophila birchii populations from tropical rainforests in Queensland, Australia. As found in a previous study, mitochondrial DNA diversity was low with no evidence for population differentiation among rainforest fragments. The pattern of mitochondrial haplotype variation was consistent with D. birchii having undergone substantial past population growth. Levels of nuclear genetic variation were high in all populations while F(ST) values were very low, even for flies from geographically isolated areas of rainforest. No significant differentiation was observed between populations on either side of the Burdekin Gap (a long-term dry corridor), although there was evidence for higher gene diversity in low-latitude populations. Spatial autocorrelation coefficients were low and did not differ significantly from random, except for one locus which revealed a clinal-like pattern. Comparisons of microsatellite differentiation contrasted with previously established clinal patterns in quantitative traits in D. birchii, and indicate that the patterns in quantitative traits are likely to be due to selection. These results suggest moderate gene flow in D. birchii over large distances. Limited population structure in this species appears to be due to recent range expansions or cycles of local extinctions followed by recolonizations/expansions. Nevertheless, patterns of local adaptation have developed in D. birchii that may result in populations showing different selection responses when faced with environmental change. PMID- 17402984 TI - Phylogenetic evidence for an animal pathogen origin of ergot and the grass endophytes. AB - Grass-associated fungi (grass symbionts) in the family Clavicipitaceae (Ascomycota, Hypocreales) are species whose host range is restricted to the plant family Poaceae and rarely Cyperaceae. The best-characterized species include Claviceps purpurea (ergot of rye) and Neotyphodium coenophialum (endophyte of tall fescue). They have been the focus of considerable research due to their importance in agricultural and grassland ecosystems and the diversity of their bioactive secondary metabolites. Here we show through multigene phylogenetic analyses and ancestral character state reconstruction that the grass symbionts in Clavicipitaceae are a derived group that originated from an animal pathogen through a dynamic process of interkingdom host jumping. The closest relatives of the grass symbionts include the genera Hypocrella, a pathogen of scale insects and white flies, and Metarhizium, a generalist arthropod pathogen. These data do not support the monophyly of Clavicipitaceae, but place it as part of a larger clade that includes Hypocreaceae, a family that contains mainly parasites of other fungi. A minimum of 5-8 independent and unidirectional interkingdom host jumps has occurred among clavicipitaceous fungi, including 3-5 to fungi, 1-2 to animals, and 1 to plants. These findings provide a new evolutionary context for studying the biology of the grass symbionts, their role in plant ecology, and the evolution of host affiliation in fungal symbioses. PMID- 17402985 TI - Combined ecological niche modelling and molecular phylogeography revealed the evolutionary history of Hordeum marinum (Poaceae)--niche differentiation, loss of genetic diversity, and speciation in Mediterranean Quaternary refugia. AB - The Hordeum marinum species group consists of two annual grasses of western Eurasian saline meadows or marshes. The two grasses split in the Quaternary about two million years ago. Hordeum marinum and the diploid of Hordeum gussoneanum (2x) co-occur throughout the Mediterranean basin, while the autotetraploid cytotype of H. gussoneanum (4x) overlaps with its diploid progenitor geographically only in the utmost Eastern Mediterranean, extending from there eastwards into Asia. Using chloroplast sequences of the trnL-F region, six newly developed chloroplast microsatellite loci, ecological predictive models based on climate data, and the present geographical distribution of the two species we analysed differentiation processes in the H. marinum group. The chloroplast data indicated clear differences in the history of both species. For H. marinum we found a subdivision between genetically variable populations from the Iberian Peninsula and the more uniform populations from the remaining Mediterranean. As an explanation, we assume Pleistocene fragmentation of an earlier widespread population and survival in an Iberian and a Central Mediterranean glacial refuge. Chloroplast variation was completely absent within the cytotypes of H. gussoneanum, indicating a severe and recent genetic bottleneck. Due to this lack of chloroplast variation only the combination of ecological habitat modelling with molecular data analyses allowed conclusions about the history of this taxon. The distribution areas of the two cytotypes of H. gussoneanum overlap today in parts of Turkey, indicating an area with similar climate conditions during polyploid formation. However, after its origin the polyploid cytotype underwent a pronounced ecological shift, compared to its diploid progenitor, allowing it to colonize mountainous inland habitats between the Mediterranean basin and Afghanistan. The extant sympatric occurrence of H. marinum and H. gussoneanum 2x in the Mediterranean region is interpreted as a result of secondary contact after fast Holocene range expansion out of different ice age refugia. PMID- 17402986 TI - Phylogeography of the white-breasted nuthatch (Sitta carolinensis): diversification in North American pine and oak woodlands. AB - Pine and oak woodlands are common North American floral communities with distinct regional species composition. The white-breasted nuthatch (Aves: Sitta carolinensis) is a common resident bird of North American pine and oak woodlands, and is distributed continentally across the highly disjunct distribution of these woodlands. We propose three historical hypotheses to explain the evolution of the white-breasted nuthatch in its principal habitat. (i) The species evolved in situ in the regional pine-oak communities and the isolation of populations in these regions is captured in cryptic genetic variation. (ii) Migration of individuals between regions is frequent enough to maintain the widespread distributions and prevent regional divergence. (iii) The species have recently expanded to occupy their current distributions and an insufficient amount of time has passed for divergence to occur. Phylogenetic analysis of mitochondrial DNA (ND2 gene) variation (N = 216) in the white-breasted nuthatch reveals four reciprocally monophyletic clades concordant with the distribution of the regional North American pine and oak woodlands, and supports hypothesis 1 of in situ evolution of populations in the regional pine and oak communities. Within-clade population structure and demographic history are also discussed. PMID- 17402987 TI - Introgression in the genus Salmo via allotriploids. AB - Hybridization between sympatric species is not uncommon in the wild. Wild allotriploids (individuals with two chromosome sets from a species + one chromosome set from another species) are generally the result of a backcross between interspecific hybrids that produce unreduced gametes and one of the parental species. In animals, allotriploids are commonly sterile, except for some vertebrate species complexes in which allotriploids reproduce by parthenogenesis, gynogenesis and/or hybridogenesis, producing generally clonal or hemiclonal gametes; nuclear DNA introgression between hybridizing species is considered to be extremely rare. Employing species-specific molecular markers, we show genetic introgression between the chromosomally well-differentiated salmonids Atlantic salmon (2n = 58) and brown trout (2n = 80) through spontaneous bisexual reproduction of allotriploids leading to salmon-like offspring bearing some brown trout genes. Although introgression between these Salmo species can occur via allotriploids, we hypothesize that extinction of parental species can be discarded based on very low survival of allotriploid offspring. PMID- 17402988 TI - Heterologous microarray experiments used to identify the early gene response to heat stress in a coral reef fish. AB - Coral reef fishes are expected to experience rising sea surface temperatures due to climate change. How well tropical reef fishes will respond to these increased temperatures and which genes are important in the response to elevated temperatures is not known. Microarray technology provides a powerful tool for gene discovery studies, but the development of microarrays for individual species can be expensive and time-consuming. In this study, we tested the suitability of a Danio rerio oligonucleotide microarray for application in a species with few genomic resources, the coral reef fish Pomacentrus moluccensis. Results from a comparative genomic hybridization experiment and direct sequence comparisons indicate that for most genes there is considerable sequence similarity between the two species, suggesting that the D. rerio array is useful for genomic studies of P. moluccensis. We employed this heterologous microarray approach to characterize the early transcriptional response to heat stress in P. moluccensis. A total of 111 gene loci, many of which are involved in protein processing, transcription, and cell growth, showed significant changes in transcript abundance following exposure to elevated temperatures. Changes in transcript abundance were validated for a selection of candidate genes using quantitative real-time polymerase chain reaction. This study demonstrates that heterologous microarrays can be successfully employed to study species for which specific microarrays have not yet been developed, and so have the potential to greatly enhance the utility of microarray technology to the field of environmental and functional genomics. PMID- 17402989 TI - Meta-analysis: mortality in Crohn's disease. AB - AIM: To perform a meta-analysis is of published literature reporting standardized mortality ratios (SMR) for Crohn's patients from 1970 to date. METHODS: Medline search identified relevant papers. Exploding references identified additional papers. When two papers reviewed mortality of one patient group at different times, the later publication was used. RESULTS: Of 13 papers identified, three studies reported SMR below 1.0, two others had confidence intervals including 1.0. All other studies reported mortality higher than the general population. Meta-analysis using a random effects model shows the pooled estimate for SMR in Crohn's disease is 1.52 (95% CI: 1.32 to 1.74 [P < 0.0001]). Meta-regression shows the SMR for these patients has decreased slightly over the past 30 years, but this decrease is not statistically significant (P = 0.08). CONCLUSION: Assessing evidence from original studies and conducting a meta-analysis shows age adjusted mortality risk from Crohn's disease is over 50% greater than the general population. Whilst mortality has improved since the condition was first recognized, further evaluation of the patients studied in the cohorts included here is necessary to assess more recent changes in clinical practice. PMID- 17402990 TI - Meta-analysis: Lactobacillus GG for treating acute diarrhoea in children. AB - AIM: To review evidence for the effectiveness of Lactobacillus GG (LGG) in treating acute infectious diarrhoea in children. METHODS: The following electronic databases were searched through August 2006 for studies relevant to acute infectious diarrhoea and LGG: MEDLINE, EMBASE, CINAHL and The Cochrane Library; additional references were obtained from reviewed articles. Only randomized-controlled trials (RCTs) were included. RESULTS: Eight RCTs (988 participants) met the inclusion criteria. Compared with controls, LGG had no effect on the total stool volume (two RCTs, n = 303). However, LGG was associated with a significant reduction in diarrhoea duration (seven RCTs, 876 infants, weighted mean difference, WMD -1.1 days (95% confidence interval, CI -1.9 to 0.3), particularly of rotavirus etiology (WMD -2.1 days, 95% CI -3.6 to -0.6), risk of diarrhoea >7 days (one RCT, n = 287, relative risk 0.25, 95% CI 0.09 0.75) and duration of hospitalization (three RCTs, n = 535, WMD -0.58, 95% CI 0.8 to -0.4; significance was lost in the random effect model). There was no reduction in the number of stools at any time interval. CONCLUSIONS: The use of LGG is associated with moderate clinical benefits in the treatment of acute diarrhoea in children. These findings should be interpreted with caution due to the important methodological limitations and heterogeneity of most of the studies. PMID- 17402991 TI - GI epidemiology: nonalcoholic fatty liver disease. AB - BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is a common diagnosis in clinical practice. Insulin resistance and oxidative stress play an important role in NAFLD development and progression. AIM: To review the data available on the epidemiology and natural history of NAFLD as well as the risk factors for its development and the areas where future research is necessary. RESULTS /CONCLUSIONS: NAFLD may affect individuals of any age range and race/ethnicity. NAFLD affects one in three adults and one in ten children/adolescents in the United States. Mortality in patients with NAFLD is significantly higher than in the general population of same age and gender with liver-related complications being a common cause of death. Liver-related morbidity and mortality in NAFLD occurs when the disease has progressed to advanced fibrosis and cirrhosis. Further studies are necessary to determine the impact of NAFLD on health-related quality of life and resources utilization, and to the extent to which preventing the development of the metabolic syndrome would prevent NAFLD development and reduce liver-related morbidity and mortality. Lifestyle intervention may improve NAFLD, but medications that increase insulin sensitivity and the antioxidant defenses in the liver deserve evaluation in carefully controlled trials. PMID- 17402992 TI - Early virological suppression is associated with good maintained response to adefovir dipivoxil in lamivudine resistant chronic hepatitis B. AB - AIM: To determine the factors affecting the virological response to adefovir dipivoxil (ADV) among patients with lamivudine resistant chronic hepatitis B. METHODS: Chronic hepatitis B virus (HBV) infected patients, who had virological relapse to lamivudine, were switched to ADV monotherapy. RESULTS: Twenty-six patients were treated by ADV for 23 (12-41) months. At baseline, the median log HBV DNA was 7.70 (4.88-9.01) copies/mL. Six (23%) and 8 (31%) of patients had HBV DNA suppressed to below 1000 copies/mL at month 12 and the last follow-up, respectively. On linear regression, patients who had higher HBV DNA at baseline and month 6 have higher HBV DNA at month 12. On Cox proportional hazard model, the hazard ratio for each log step increase in HBV DNA at baseline and month 6 for HBV DNA <1000 copies/mL at the last visit was 0.39 (P = 0.010) and 0.47 (P = 0.027), respectively. Alanine aminotransferase, HBV genotype, rtL80 M mutation and log HBsAg did not affect the HBV DNA response. CONCLUSIONS: The response of lamivudine-resistant patients to ADV is suboptimal. Treatment with ADV when HBV DNA is low, and rapid viral suppression at month 6 increases the chance of maintained viral suppression. PMID- 17402993 TI - Treatment of chronic he1patitis C genotype 1 with peginterferon-alpha2a (40 kDa) plus ribavirin under routine clinical practice in Spain: early prediction of sustained virological response rate. AB - BACKGROUND: Sustained virological response rates of up to 52% have been obtained with peginterferon alpha2a (40 kDa) plus ribavirin in patients suffering from chronic hepatitis C genotype 1 in randomized-controlled trials. AIM: To assess early virological response and its clinical utility in predicting an sustained virological response in patients suffering from chronic hepatitis C genotype 1 in routine clinical practice in Spain. METHODS: Treatment-naive patients received pegylated interferon alpha2a (40 kDa) 180 microg/week plus ribavirin 1000/1200 mg/day for 48 weeks, and were followed for a further 24 weeks. Overall, 475 patients received at least one dose of medication and were included in the efficacy population. RESULTS: The overall sustained virological response rate was 48%. Of those with week 12 virological data, 83% had an early virological response. The negative predictive value of an early virological response was 93%. CONCLUSION: If sustained virological response is the goal, a treatment-decision based on a 12-week evaluation during routine clinical practice is feasible. PMID- 17402994 TI - Introduction of newer selective cyclo-oxygenase-2 inhibitors and rates of hospitalization with bleeding and perforated peptic ulcer. AB - BACKGROUND: Limited data exist on the impact of the introduction of newer selective cyclo-oxygenase-2 inhibitors into clinical practice in 1999 on overall non-steroidal anti-inflammatory drug use and hospitalization rates of complicated peptic ulcer disease at the population level. AIM: To examine these issues, we conducted a population-based study in western Denmark, within a population of 1.2 million. METHODS: All patients with perforated (n = 1488) or bleeding peptic ulcer (n = 6017) between 1996 and 2004 were identified in hospital discharge registries. We computed standardized annual hospitalization rates and hospitalization rate ratios using Poisson regression. Data on annual number of prescriptions for non-steroidal anti-inflammatory drugs were obtained through population-based prescription databases. Results Introduction of newer selective cyclo-oxygenase-2 inhibitors was followed by a 44% increase in the annual number of prescriptions for non-steroidal anti-inflammatory drugs--almost entirely due to increased use of newer selective cyclo-oxygenase-2 inhibitors. Annual standardized hospitalization rates for bleeding peptic ulcer remained stable. Standardized hospitalization rates for perforated peptic ulcer decreased from 17 per 100,000 person-years in 1996 to 12 per 100,000 person-years in 2004 (HRR 0.71; 95% CI: 0.57-0.88). Conclusion Introduction of newer selective cyclo oxygenase-2 inhibitors was followed by substantial increase in overall non steroidal anti-inflammatory drug use and coincided with stable and decreasing hospitalization rates for bleeding and perforated peptic ulcer, respectively. PMID- 17402995 TI - Cyclooxygenase-2 selectivity of non-steroidal anti-inflammatory drugs and the risk of myocardial infarction and cerebrovascular accident. AB - AIM: To assess degree of cyclooxygenase-2 (COX-2) selectivity of a non-steroidal anti-inflammatory drug (NSAID) and risk of myocardial infarction (MI) or cerebrovascular accident (CVA). METHODS: Prescription fill data were linked to medical records of a merged VA-Medicare dataset. NSAIDs were categorized by Cox-2 selectivity. Incidence of CVA and MI within 180 days of index prescription was assessed using Cox-proportional hazards models adjusted for gender, race, cardiovascular and pharmacological risk factors and propensity for prescription of highly COX-2 selective NSAIDs. RESULTS: Of 384,322 patients (97.5% men and 85.4% white), 79.4% were prescribed a poorly selective, 16.4% a moderately selective and 4.2% a highly selective NSAID. There were 985 incident cases of MI and 586 cases of CVA in >145 870 person-years. Highly selective agents had the highest rate of MI (12.3 per 1000 person-years; [95% CI: 12.2-12.3]) and CVA (8.1 per 1000 person-years; [95% CI: 8.0-8.2]). Periods without NSAID exposure were associated with lowest risk. In adjusted models, highly selective COX-2 selective NSAIDs were associated with a 61% increase in CVA and a 47% increase in MI, when compared with poorly selective NSAIDs. CONCLUSIONS: The risk of MI and CVA increases with any NSAID. Highly COX-2 selective NSAIDs confer the greatest risk. PMID- 17402996 TI - Early changes in rectal nitric oxide and mucosal inflammatory mediators in Crohn's colitis in response to infliximab treatment. AB - BACKGROUND: Treatment with tumor necrosis factor-alpha monoclonal antibody (infliximab) reduces clinical activity and intestinal inflammation in Crohn's disease. AIM: To study the time-course of the effects of infliximab with reference to mucosal cytokine and inducible nitric oxide synthase expression. METHODS: Thirty-two patients with Crohn's disease were treated with single dose infliximab (5 mg/kg). Disease activity was assessed days 1, 3, 7 and 28 using Harvey-Bradshaw index. Rectal nitric oxide levels were determined and rectal biopsies collected before treatment, 1 h after infusion and on days 3, 7 and 28. Immunohistochemical staining against inducible nitric oxide synthase, tumor necrosis factor-alpha, interleukin-1beta and interferon-gamma were performed. RESULTS: Clinical response was seen in 14 patients with down-regulation of global immunohistochemistry expression, reaching nadir day 3. Rectal nitric oxide was increased at baseline (3578 +/- 1199 parts per billion, ppb) compared with controls (89 +/- 13 ppb) (P < 0.001). In patients with clinical response, rectal nitric oxide decreased from 3926 +/- 1687 ppb to 1050 +/- 428 ppb day 28 (P < 0.05). CONCLUSIONS: Down-regulation of mucosal inflammatory mediators occurs after infliximab. Rectal nitric oxide levels parallel down-regulation of inducible nitric oxide synthase, tumor necrosis factor-alpha, interleukin-1beta and interferon-gamma and may serve as a quantitative biomarker of intestinal inflammation. PMID- 17402997 TI - Altered intestinal permeability is predictive of early relapse in children with steroid-responsive ulcerative colitis. AB - AIM: To determine if small bowel involvement at diagnosis could predict early relapse in children with ulcerative colitis. METHODS: Children with newly diagnosed ulcerative colitis were evaluated prospectively at three time points: within 1 month, 6 months and 1 year after diagnosis. Clinical activity indices were used to measure disease activity. Laboratory studies were performed at each visit and/or at the time of relapse. At diagnosis, all patients underwent colonoscopy and a cellobiose/mannitol small intestinal permeability study. Some children were further investigated with an upper gastrointestinal endoscopy. RESULTS: Thirty-three patients completed the 1-year study. Overall, nine patients (27.3%) relapsed within 6 months of diagnosis, one patient (3%) within 1 year, whereas 23 patients (69.7%) did not relapse. The mean clinical activity indices, laboratory parameters, extent of colonic involvement, upper and lower gastrointestinal histological features were not predictive of early relapse. Results of the cellobiose/mannitol small intestinal permeability study were significantly higher in children who relapsed within 6 months compared with children who did not relapse (P < 0.013). The cellobiose/mannitol small intestinal permeability study was abnormal in 77.8% of early relapsers compared with only 8.3% of non-relapsers. CONCLUSION: Abnormal small intestinal permeability in children with ulcerative colitis could predict a more relapsing disease. PMID- 17402998 TI - Thiopurine metabolite monitoring in paediatric inflammatory bowel disease. AB - BACKGROUND: Measurement of thiopurine metabolite levels may be useful as a clinical tool to optimize thiopurine treatment of paediatric inflammatory bowel disease (IBD). AIM: The authors evaluated correlations between 6-thioguanine nucleotide (6-TGN) and therapeutic response, metabolite levels and drug toxicity. METHODS: Fifty-six paediatric IBD patients treated with thiopurines had 326 metabolite level measurements and were retrospectively reviewed. Clinical status and laboratory parameters were compared with metabolite levels. RESULTS: There was significant correlation between 6-TGN levels and therapeutic response, with higher median 6-TGN levels among patients with therapeutic response than those with non-therapeutic response (194 vs. 146 pmol/8 x 10(8) RBC; P = 0.0004). Patients with 6-TGN levels >235 pmol/8 x 10(8) RBC were more likely to achieve therapeutic response than those below the cut-off (odds ratio, 2.5; 95% CI, 1.5 4.1). Patients who developed leukopenia tended to have higher median 6-TGN levels than those without leukopenia (261 vs. 160 pmol/8 x 10(8) RBC) but the difference was not statistically significant. There was no correlation between 6 methylmercaptopurine levels and hepatotoxicity. Two patients developed acute pancreatitis. Metabolite level measurements were helpful in identifying non compliance in nine patients. CONCLUSIONS: Monitoring of thiopurine metabolite levels is useful to guide and optimize dosing, as an adjunct to clinical judgement, blood count and liver biochemistry measurements to minimize the risk of drug toxicity and to confirm non-compliance. PMID- 17402999 TI - Continuous infusion of high-dose omeprazole is more effective than standard-dose omeprazole in patients with high-risk peptic ulcer bleeding: a retrospective study. AB - High-dose omeprazole reduces the rate of recurrent bleeding after endoscopic treatment of peptic ulcer bleeding. However, the effectiveness of high-dose vs. standard-dose omeprazole in peptic ulcer bleeding has never been shown. AIM: To compare the benefits of high-dose vs. standard-dose omeprazole in peptic ulcer bleeding. METHODS: We reviewed the medical files of patients admitted between 1997 and 2004 for high-risk peptic ulcer bleeding who had undergone successful endoscopic treatment. We distinguished 2 periods: before 2001, standard-dose omeprazole (40 mg/day intravenously until alimentation was possible, then 40 mg/day orally for 1 week); after 2001, high-dose omeprazole (80 mg bolus injection, then 8 mg/h continuous infusion for 72 h, then 40 mg/day orally for 1 week). During both periods, patients subsequently received omeprazole, 20 mg/day, orally for 3 weeks. RESULTS: We enrolled 114 patients (period 1, n = 45, period 2, n = 69). Therapy with high-dose omeprazole significantly decreased the occurrence of poor outcome (27 vs. 12%, P = 0.04), rebleeding (24 vs. 7%, P = 0.01), mortality due to haemorrhagic shock (11 vs. 0%, P < 0.001) and need for surgery (9 vs. 1%, P = 0.05). CONCLUSIONS: In this retrospective study, high-dose omeprazole reduced the occurrence of rebleeding, need for surgery and mortality due to hemorrhagic shock in patients with high-risk peptic ulcer bleeding, as compared with standard-dose omeprazole. PMID- 17403000 TI - Double stenting of oesophagus and airways in palliative treatment of patients with oesophageal cancer is efficient but associated with a high morbidity. AB - BACKGROUND: Double stenting of oesophagus and airways may be required in palliative treatment of patients with locally advanced oesophageal cancer. AIM: To assess feasibility, efficacy and complications occurring in patients with locally advanced oesophageal cancer receiving both oesophagus and airways stenting. METHODS: In one single centre between 1997 and 2005, among 180 patients with locally advanced oesophageal cancer treated by the palliative placement of a self-expanding metal stent, patients requiring double stenting of oesophagus and airways were identified. Clinical efficacy, complications and survival were retrospectively collected. RESULTS: Fifteen patients (8.3% of 180) required a double stenting at follow-up. Symptomatic efficacy of oesophagus and airways stenting was 86.7% for dysphagia and 100% for dyspnoea. Median survival after the second stent insertion was 99 days. Life-threatening early complications occurred in three patients after double stenting (20%), including two deaths following oesophageal perforation and massive haemoptysis, respectively. Procedure-related mortality was 13.3%. CONCLUSIONS: Double stenting of oesophagus and airways is feasible in patients with locally advanced oesophageal cancer, with a relevant clinical efficacy. However, early major complications including procedure-related death may occur in as many as 20% of patients. This treatment should be reserved to very selected patients with severe symptoms and end-stage disease. PMID- 17403001 TI - Air swallowing, belching, acid and non-acid reflux in patients with functional dyspepsia. AB - BACKGROUND: Frequent belching is a common symptom in patients with functional dyspepsia with a reported incidence up to 80%. We hypothesized that patients with functional dyspepsia possibly have a higher frequency of belching than healthy subjects secondary to frequent air swallowing. AIM: To assess air swallowing, belching, acid and non-acid reflux patterns of patients with functional dyspepsia. METHODS: Combined 24-h oesophageal impedance and pH monitoring was performed in 10 functional dyspepsia patients and 10 controls. Analysis of the impedance-pH signals included incidence of air swallows, belching, acid and non acid reflux. RESULTS: The incidence of air swallows in functional dyspepsia patients were significantly higher compared with controls (153 +/- 15 vs. 79 +/- 10, P < 0.001), while the incidence of liquid-only swallows were not significantly increased. The proportions of gas-containing reflux episodes (belches) and non-acid reflux episodes in functional dyspepsia patients were significantly higher when compared with controls (66.4 vs. 44.4%, P = 0.04 and 70.1 vs. 45.9%, P = 0.009, respectively). CONCLUSION: Patients with functional dyspepsia swallow air more frequently than controls and this is associated with an increased incidence of non-acid gaseous gastro-oesophageal reflux. PMID- 17403002 TI - Is there a benefit from intensified medical and psychological interventions in patients with functional dyspepsia not responding to conventional therapy? AB - AIM: In a prospective randomized, controlled trial, to compare the long-term outcome of intensive medical therapy (with or without cognitive-behavioural or muscle relaxation therapy) vs. standard medical therapy in patients with refractory functional dyspepsia (FD), referred to a tertiary referral medical center. METHODS: A total of 100 consecutive FD patients were allocated to a standardized symptom-oriented 4 month therapy (SMT, n = 24), intensive medical therapy (IMT, medical therapy with testing-for and targeting-of abnormalities of motor-and-sensory function, n = 28) or IMT plus psychological interventions (either progressive-muscle relaxation (IMT-MR, n = 20) or cognitive-behavioural therapy (IMT-CBT, n = 28). The symptom intensity (SI) and health-related quality of-life (HRQoL) after 12 months were prespecified primary outcome parameters. RESULTS: After 12 months, significantly greater improvement of SI occurred in patients with IMT-all (with or without psychological interventions) compared with SMT (P < 0.025 vs. IMT-all). IMT, IMT-MR and IMT-CBT alone also resulted in significantly better improvement of the primary outcome parameters (P all < 0.025 vs. SMT). HRQoL significantly improved in all groups with intensive medical therapy but not standard medical therapy. Differences between intensive medical therapy-all and standard medical therapy were not significant. Concomitant anxiety and depression was improved significantly by IMT-CBT (vs. SMT) but not other treatments. CONCLUSIONS: In FD patients with refractory symptoms, intensified medical management involving function testing and psychological intervention yields superior long-term-outcomes. Additional CBT may be effective for the control of concomitant anxiety and depression. PMID- 17403003 TI - Prospective randomized trial evaluating ketamine for advanced endoscopic procedures in difficult to sedate patients. AB - BACKGROUND: Adequate patient sedation is mandatory for advanced endoscopic procedures such as ERCP and EUS. AIM: To evaluate the effectiveness and safety of ketamine in difficult to sedate patients undergoing advanced endoscopic procedures. METHODS: This was a prospective, randomized trial of all patients undergoing ERCP or EUS who were not adequately sedated despite administration of meperidine 50 mg, midazolam 5 mg and diazepam 5 mg. Patients during endoscopy were then randomized to receive either intravenous ketamine (20 mg) every 5 min or continue to receive standard sedation using meperidine and diazepam. RESULTS: Of 175 patients, 82 were randomized to receive ketamine and 93 standard sedatives. Compared with standard sedation, qualitative physician rating (P < 0.0001) and depth of sedation (P < 0.001) were superior in the ketamine group with shorter recovery times (P < 0.0001). Both patient discomfort and sedation related technical difficulty were significantly less among patients randomized to receive ketamine (P < 0.0001). More patients in the standard sedation group were crossed-over to the ketamine group due to sedation failure (35.5 vs. 3.7%, P < 0.0001). Nine patients who received ketamine, developed adverse events that were managed conservatively. CONCLUSIONS: Ketamine is a useful adjunct to conscious sedation in patients who are difficult to sedate. Its use Results in better quality and depth of sedation with shorter recovery times than patients sedated using benzodiazepines and meperidine alone. Further prospective studies evaluating the effectiveness and safety of ketamine for endoscopic sedation are needed. PMID- 17403004 TI - Rebound acid hypersecretion. PMID- 17403008 TI - Genetic mapping of quantitative trait loci affecting carcass and meat quality traits in Beijing ducks (Anas platyrhynchos). AB - Quantitative trait loci (QTL) for carcass and meat quality traits were detected in a sample of 224 progeny from four males in line VI and 12 females in line V of Beijing ducks. These lines were selected for high body weight at 42 days of age (line VI) or high egg production at 360 days of age (line V). Traits were weights of the carcass, head, neck, shanks, wings, legs, thighs, breast, heart, liver, crop, gizzard, abdominal fat (AFW) and skin fat, as well as fat thickness in the tail, and pH value, shear force, drip loss (DL) (%) and cooking loss (CL) (%) of the breast. Using a half-sib analysis with a multiple QTL model, linkage between the carcass and meat quality traits and 95 microsatellite markers was investigated. Eight genome-wide significant QTL for weight of crop, skin fat, liver, neck, shanks, wings, DL were detected on linkage groups CAU4 and CAU6. One genome-wide suggestive QTL and one chromosome-wide significant QTL for weight of breast were found on CAU1 and CAU4 respectively. Fifteen chromosome-wide suggestive QTL influencing weight of AFW, breast, crop, heart, carcass, thighs, liver, shanks, gizzard, fat thickness in tail, DL (%) and CL (%) were mapped on CAU2, CAU4, CAU5, CAU6, CAU7, CAU10 and CAU13. In addition, two linked QTL for weight of liver and DL (%) were located on CAU2 and CAU7 respectively. The detection of QTL in ducks is a step towards identification of genes influencing these traits and their use for genetic improvement in this species. PMID- 17403009 TI - Cloning, mapping and association studies of the ovine ABCG2 gene with facial eczema disease in sheep. AB - Facial eczema (FE) is a hepatogenous mycotoxicosis in sheep caused by the fungal toxin sporidesmin. Resistance to FE is a multigenic trait. To identify QTL associated with this trait, a scan of ovine chromosomes was implemented. In addition, ABCG2 was investigated as a possible positional candidate gene because of its sequence homology to the yeast PDR5 protein and its functional role as a xenobiotic transporter. The sequence of ovine ABCG2 cDNA was obtained from liver mRNA by RT-PCR and 5' and 3' RACE. The predicted protein sequence shares >80% identity with other mammalian ABCG2 proteins. SNPs were identified within exon 6, exon 9 and intron 4. The intron 4 SNP was used to map ABCG2 to ovine chromosome 6 (OAR6), about 2 cM distal to microsatellite marker OarAE101. Interestingly, this chromosomal region contains weak evidence for a FE QTL detected in a previous genome-scan experiment. To further investigate the association of ABCG2 with FE, allele frequencies for the three SNPs plus three neighbouring microsatellite markers were tested for differences in sheep selected for and against FE. Significant differences were detected in the allele frequencies of the intronic SNP marker among the resistant, susceptible and control lines. No difference in the levels of ABCG2 expression between the resistant and susceptible animals was detected by Northern hybridisation of liver RNA samples. However, significantly higher expression was observed in sporidesmin-dosed sheep compared with naive animals. Our inference is that the ABCG2 gene may play a minor role in FE sensitivity in sheep, at least within these selection lines. PMID- 17403010 TI - IGF2 gene characterization and association with rib eye area in beef cattle. AB - Insulin-like growth factor 2 (IGF2) is an imprinted gene expressed in most tissues affecting lean muscle content in mice, pigs and cattle. We previously identified the bovine IGF2 c.-292C>T SNP in the non-translated exon 2. Using this SNP, we demonstrated biallelic expression of IGF2 after birth. Seven alternatively spliced mRNA transcripts of IGF2 were expressed among 15 tissues. An IGF2 pseudogene (psiIGF2) was identified with sequence identical to at least IGF2 exons 2 and 3 without the intervening intron. The biallelic expression of this c.-292C>T SNP was associated with an increase in rib eye area (REA) in two populations of cattle, with the C.-292C allele associated with a 10% increase. A significant association with per cent fat was found in one of the populations. PMID- 17403015 TI - Obesity and periodontitis in 60-70-year-old men. AB - OBJECTIVE: The aim was to investigate whether there was an association between obesity and periodontitis in a homogeneous group of 60-70-year-old Western European men. The study also explored whether a high body mass index (BMI) in early life predicted poor periodontal status in later life. METHODOLOGY: A representative sample of the male population of UK, who were enrolled in a cohort study of cardiovascular disease, was examined between 2001 and 2003. A total of 1362 men with six or more teeth completed a questionnaire, had a clinical periodontal examination and had their weight and height recorded. Multivariable analysis was carried out using logistic regression with adjustment for possible confounders. Models were constructed with low- and high- threshold periodontitis as dependent variables and with BMI as a categorical predictor variable. Low threshold periodontitis was identified when at least two teeth had >/=6 mm loss of attachment and at least one site had a pocket of >/=5 mm. High-threshold periodontitis was identified when >/=15% of sites had >/=6 mm loss of attachment and there was deep pocketing (>/=6 mm). Modelling was repeated for other predictor variables including BMI at 21 years of age and change in weight since 21 years of age. RESULTS: Two hundred and ninety-eight (21.9%) of the men studied, who had a BMI of >30 kg/m(2), were classified as obese. Obesity was associated with low-threshold periodontitis, odds ratio (OR)=1.77 (p=0.004) after adjustment for confounders. The BMI at 21 years of age did not predict periodontitis in the men investigated. Participants who had experienced a large (>30%) increase in weight during adulthood had an increased risk of poor periodontal condition; however, this was attenuated and no longer significant after adjustment for confounders. CONCLUSIONS: It is concluded that obesity was associated with periodontitis in the homogeneous group of 60-70-year-old European men investigated. High BMI levels in early life did not predict periodontitis in later life in the men studied. PMID- 17403016 TI - Management of smoking patients by specialist periodontists and hygienists in the United Kingdom. AB - OBJECTIVE: To assess the management of patients who are current smokers in dental practice. MATERIALS AND METHODS: A postal questionnaire to the 243 periodontists on the UK specialist register and 239 randomly selected hygienists on the General Dental Council register. RESULTS: Eighty per cent of both groups responded. There were significant demographic differences between the two groups of respondents: 88% of periodontists were in specialist practice and/or hospital or university departments, while 89% of hygienists were in general dental practice. Periodontists routinely asked their patients about smoking 99% of the time, hygienists 89%. More than 5 min. was spent advising a smoker patient by 35% of periodontists and 19% of hygienists. A proportion of both groups claimed to have had some recognized training in quit smoking (44% and 39%). When asked about the oral conditions of their smoking patients, 26% of periodontists stated that these patients did not have poorer oral hygiene, while 67% of hygienists thought that they did. While the majority of both groups discussed the oral effects of smoking, less than two-thirds discussed methods of giving up. CONCLUSION: Both periodontists and hygienists are generally aware of the problems of treating smokers, and accordingly ask and advise their patients. However, lack of time and poor patient response may inhibit active involvement in assisting patients to quit smoking. PMID- 17403017 TI - Opposite effects of TGF-beta1 and IFN-gamma on transdifferentiation of myofibroblast in human gingival cell cultures. AB - BACKGROUND/AIM: Previously, we have shown that myofibroblasts, the main cell type associated with interstitial fibrosis, may be implicated with the gingival overgrowth observed in hereditary gingival fibromatosis (HGF) patients. The goal of this study was to determine whether transforming growth factor-beta1 (TGF beta1) stimulates myofibroblast generation in gingival fibroblast cultures. Moreover, we analysed how interferon-gamma (IFN-gamma) interferes in this process. MATERIAL AND METHODS: Fibroblast cultures from normal gingiva and myofibroblast cells from HGF were included in this study. To determine the effects of TGF-beta1 and IFN-gamma stimulation in these cells, the expression of the specific myofibroblast marker smooth muscle isoform of alpha-actin (alpha SMA) was examined by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and immunofluorescence. Enzyme-linked immunosorbent assay (ELISA) for type I collagen was performed to measure the myofibroblast activity. RESULTS: Our results demonstrated that TGF-beta1 promotes a dose- and time-dependent increase in the expression of alpha-SMA, whereas IFN gamma blocks it and markedly prevents the fibroblast-myofibroblast switch induced by TGF-beta1 on normal gingiva cultures. IFN-gamma altered HGF myofibroblasts metabolism with a decrease of both alpha-SMA and type I collagen expression. Additionally, IFN-gamma treatment stimulated SMAD7 expression and inhibited connective tissue growth factor, which has been considered a key molecule to promote the transdifferentiation of myofibroblasts via TGF-beta1 activation. CONCLUSIONS: These findings demonstrate that TGF-beta1 induces gingival fibroblast-myofibroblast transdifferentiation, whereas IFN-gamma blocks this process. More importantly, this study suggests that IFN-gamma may be clinically effective in attenuating excessive accumulation of extracellular matrix produced by myofibroblasts in HGF. PMID- 17403018 TI - Corneal oedema and acute anterior uveitis after two doses of travoprost. PMID- 17403019 TI - Photopic 30 Hz flicker electroretinography predicts ocular neovascularization in central retinal vein occlusion. AB - PURPOSE: To confirm the predictive value of photopic cone b-wave implicit time in 30 Hz flicker electroretinography (ERG) for ocular neovascularization (NV) in central retinal vein occlusion (CRVO), and to compare the ERG results to the presumed healthy fellow eye. METHODS: A retrospective analysis of 71 consecutive patients with CRVO. After ERG examination, all patients were followed for at least 12 months, or until NV was found. Three patients died during the study period; none of the other patients were lost to follow-up. RESULTS: Twenty-four patients (33.8%) developed NV during follow-up. The mean cone b-wave implicit time of all patients was 32.6 ms [standard deviation (SD) 5.21]. All 18 patients with an implicit time of 35.0 ms or higher (> 0.5 SD from mean) developed NV. In patients who developed NV, the average implicit time was 38.5 ms (range 29.7-43.9 ms); in patients without NV (n = 47), the average implicit time was 29.6 ms (range 24.7-34.9 ms) (P < 0.0001). The average implicit time in the presumed healthy fellow eye was 28.7 ms (range 24.4-33.9 ms) in patients with NV, and 26.5 ms (range 23.7-33.2 ms) in patients without NV (P = 0.002). The mean interocular difference in implicit time was 9.9 ms (range 4.1-15.7 ms) in patients with NV and 2.9 ms (range -1.0 to 10.0 ms) in patients without NV (P < 0.0001). CONCLUSION: Patients with CRVO should be examined routinely with photopic 30 Hz flicker ERG, which is a simple and objective clinical test that can identify patients at risk of ocular NV. On the assumption that the presented ERG settings are used, implicit times of 35.0 ms or higher (> 0.5 SD from mean) are clearly associated with the development of ocular NV. To compare the ERG result of the affected eye to the presumed healthy fellow eye is probably of less value. PMID- 17403020 TI - High infusion pressure in conjunction with vitreous surgery alters the morphology and function of the retina of rabbits. AB - PURPOSE: To investigate the effects of high infusion pressure in conjunction with pars plana vitrectomy (PPV) on retinal morphology and function in rabbits. METHODS: Pars plana vitrectomy was performed under urethane (0.8 mg/kg) anaesthesia in the right eye of albino rabbits following phacoemulsification and aspiration (PEA). The left eyes were not touched. After PEA, the animals were divided into two groups. In six eyes, intraocular pressure (IOP) was increased to 80 mmHg for 30 mins (high-pressure group) and in five eyes IOP was maintained at 40 mmHg for 30 mins (low-pressure group). The IOPs were regulated by the height of the bottle of balanced salt solution (BSS) and monitored with a pressure transducer. After the pressure elevation, vitreous fluid was collected to measure the glutamate concentration. Then, PPV was performed for 15 mins in both groups under an infusion pressure of 40 mmHg. In five additional rabbits, PEA alone was performed in the right eye, and vitreous fluid was collected (PEA group). Functional alterations were assessed by recording visual evoked potentials (VEPs) and electroretinograms (ERGs). Ten days after the IOP changes, the animals were killed with intravenous pentobarbital sodium and the eyes were prepared for histological analysis. Damage to retinal ganglion cells (RGCs) was quantified by counting the number of cells in the ganglion cell layer (GCL). The contralateral eyes in the high-pressure group served as controls (n = 6). RESULTS: The mean implicit time (IT) of the VEPs in the high-pressure group was significantly longer than that before the IOP elevation, by 114-124% (p < 0.05, paired t-test), and also than that of control eyes (p < 0.05, anova followed by t-test). No significant changes in the VEPs were detected in either the low-pressure group or the PEA group. There were significantly fewer cells in the GCL in the high pressure group (24.7/mm) than in the control animals (41.4/mm; p < 0.05, Dunnett's test). The number of cells in the GCL in the low-pressure and PEA groups did not significantly differ to that in the controls. The amplitudes of the ERG a- and b-waves were not significantly changed (p > 0.05, paired t-test). CONCLUSIONS: These results suggest that high infusion pressure in conjunction with PPV leads to morphological and functional changes in the retina. The absence of ERG changes and presence of VEP changes suggest that these changes were due to damage to RGCs, which supports the morphological observations. PMID- 17403021 TI - Tilted disc syndrome and colour vision. AB - PURPOSE: To study colour vision in patients with tilted disc syndrome. METHODS: Colour vision was examined using Bostrom-Kugelberg (B-K) plates, Farnsworth Munsell 100-hue test (FM-100) and Farnsworth panel D-15 and Lanthony desaturated panel tests. RESULTS: A total of 35 eyes of 21 patients were examined. Seventeen eyes (49%) of 11 patients were found to have a colour vision defect in at least one eye. The severity of the colour vision defect differed between eyes for six of eight patients with bilateral tilted discs. In two of these patients, colour vision was normal in one eye. Seven patients had a unilateral tilted disc and three of these individuals had a colour vision defect in the affected eye. Of all eyes with a colour vision defect, a red-green defect was found in four eyes (24%), a blue defect in one eye (6%), and a mixed defect in 12 eyes (70%). Colour vision defects were not related to visual acuity or to severity of visual field defects. CONCLUSIONS: This is the first report of colour vision defects in tilted disc syndrome. In the differential diagnosis of tilted disc syndrome, chiasmal lesions and glaucoma, the examination of colour vision may add valuable information. Further, it is recommended that young people with tilted disc syndrome undergo an examination of colour vision so that they can be given appropriate career guidance. PMID- 17403022 TI - Cytomegalovirus retinitis following intravitreal injection of triamcinolone: report of two cases. AB - CONTEXT: Cytomegalovirus (CMV) retinitis usually affects severely immunosuppressed individuals. However, some exceptional and isolated cases of CMV retinitis have been described in immunocompetent patients. METHODS: We report two cases of non-immunodeficient patients seen in our institution over a period of 6 months, who presented with CMV retinitis a few weeks after an intravitreal injection of triamcinolone acetonide. RESULTS: Although the cellular and molecular mechanisms underlying the development of CMV in an immunocompetent patient's eye have not yet been precisely established, the well known immunosuppressive properties of corticosteroids suggest that triamcinolone probably plays a role in the endogenous reactivation of latent CMV. CONCLUSIONS: Our two patients received high doses of triamcinolone. Both had also pseudophakia, vitrectomy and a past medical history of type 2 diabetes mellitus. Such features, individually or combined, may be risk factors for local reactivation of CMV and should be considered prior to the administration of triamcinolone acetonide in ophthalmology. PMID- 17403023 TI - Longterm oculomotor and visual function in spina bifida cystica: a population based study. AB - PURPOSE: To document and describe the development from birth of visual and oculomotor functions in a group of children with spina bifida cystica (myelomeningocele and myeloschisis [MMC]). The emphasis in this study is on findings at 12-14 year follow-up. METHODS: Twenty children aged 12-14 years with myelomeningocele and Chiari-related malformations were examined by an orthoptist and a paediatric ophthalmologist. A further child who did not wish to participate actively in the study is also reported. Visual acuity for near and distance, refractometer readings in cycloplegia, the presence of ocular motility disorders and nystagmus were recorded. Accommodation, convergence, colour vision and stereo acuity were assessed and the fundus and media were examined. RESULTS: Six children (29%) in the study group had subnormal vision, although no child was visually impaired. Eleven (52%) showed manifest strabismus and 17 (81%) had a significant refractive error. Near visual acuity was normal in nearly all the children, but accommodation was defective in 10. Nine children had nystagmus and two had optic atrophy. No visual field defects were found. CONCLUSIONS: The high incidence of ocular disturbances in children with spina bifida highlights the importance of regular ophthalmological investigation and follow-up. PMID- 17403024 TI - Calculation of intraocular lens power: a review. AB - This review describes the principles and practices involved in the calculation of intraocular lens (IOL) power. The theories behind formulas for calculating IOL power are described, using regression and optical methods employing 'thin lens' and 'thick lens' models, as well as exact ray-tracing methods. Numerical examples are included to illustrate the points made. The paper emphasizes the importance of establishing an accurate estimation of corneal power as well as an accurate technique for the measurement of axial length and accurate methods of predicting postoperative anterior chamber depth (ACD). It is concluded that current improvements in diagnostic and surgical technology, combined with the latest generation IOL power formulas, make the calculation and selection of appropriate IOL power among the most effective tools in refractive surgery today. PMID- 17403025 TI - Acute retinal pigment epithelial tear in the untreated fellow eye following repeated bevacizumab (Avastin) injections. PMID- 17403026 TI - Longterm results of 1 CU accommodative intraocular lens implantation: 2-year follow-up study. AB - PURPOSE: To evaluate the longterm efficacy of 1 CU accommodative intraocular lenses (IOLs) to restore near visual performance. METHODS: This prospective study comprised 14 eyes previously included in a 6-month, case-control clinical trial, undergoing phacoemulsification and implantation of a 1 CU accommodative IOL. The main outcome measures were subjective refraction, uncorrected distance visual acuity (UCDVA), best corrected distance VA (BCDVA), distance-corrected near VA (DCNVA), best corrected near VA (BCNVA), and subjective amplitude of accommodation (AA). In addition, anterior and posterior capsule opacification were assessed. Patients were examined over a 2-year follow-up period. RESULTS: Distance and near visual performance worsened after 6 months. Uncorrected DVA and BCDVA were 0.8 +/- 2.1 and 1.0 +/- 0.8 at 6 months and 0.4 +/- 0.1 and 0.6 +/- 0.1 at 1 year, respectively (p = 0.001). Distance-corrected NVA and BCNVA were 3.7 +/- 2.1 Jaeger (J) and 1.0 +/- 0.7 J at 6 months and 8.1 +/- 0.7 J and 1.5 +/ 0.5 J at 1 year, respectively (p = 0.001). Anterior and posterior capsule opacification were present, respectively, in 28% and 21% of patients at 6 months and in 100% of patients at 1 and 2 years (p < 0.001). After Nd:YAG laser capsulotomy (performed in 100% of patients), UCDVA and BCDVA increased to 0.7 +/- 0.2 (p = 0.007) and 1.0 +/- 0.1 (p = 0.001), respectively, at 2 years. Distance corrected NVA improved to 7.3 +/- 0.5 J (p = 0.006). Mean AA was 1.9 +/- 0.8 D at 6 months, 0.3 +/- 0.2 D (p = 0.004) at 1 year and 0.3 +/- 0.2 D at 2 years. CONCLUSIONS: Patients implanted with 1 CU IOLs lost their accommodation capacities with time because of the high incidence and degree of anterior and posterior capsule opacification. The accommodative lens material and design may have played a role in capsule fibrosis. PMID- 17403027 TI - The use of meta-analysis risk estimates for candidate genes in combination to predict coronary heart disease risk. AB - Although the risk for coronary heart disease (CHD) associated with single SNPs is modest it has been suggested that, in combination, several common risk-associated alleles could lead to a substantially better heart disease risk prediction. We have modelled this using 10 SNPs in ten candidate genes (APOB, NOS3, APOE, ACE, SERPINE1, MTHFR, ITGA2B, PON 1, LPL, and CETP) and their predicted summary risk estimates from meta-analysis. Based on published allele frequencies, approximately 29% of the general population would be expected to carry less than three risk alleles, approximately 55% would carry 3 or 4 risk alleles, 4% would have 6 and 1% 7 or more risk alleles. Compared to the mean of those with 3 or 4 risk associated genotypes, those with 6 and 7-or-more alleles have a significantly higher risk odds ratio (OR) of CHD (mean OR (95% Confidence Intervals), 1.70 (1.14 to 2.55); and 4.51 (2.89 to 7.04) respectively), while compared to those in the lowest decile of risk, those in the highest decile have a CHD odds ratio in the range of 3.05 (2.24 to 4.14). Taking into account age and the risk alleles carried, the mean 10 year probability for developing CHD for a 55 year old man was calculated to be 15% (8.6% to 24.8%), with nearly 1 in 5 having more than 20% risk. Whether this particular group of 10 SNPs will improve the accuracy of CHD predictions over the combination of classical risk factors in clinical use requires further experimental evidence. PMID- 17403028 TI - Two isoforms of the cold-inducible mRNA-binding protein RBM3 localize to dendrites and promote translation. AB - A diverse set of mRNA-binding proteins (BPs) regulate local translation in neurons. However, little is known about the role(s) played by a family of cold inducible, glycine-rich mRNA-BPs. Unlike neuronal mRNA-BPs characterized thus far, these proteins are induced by hypothermia and are comprised of one RNA recognition motif and an adjacent arginine- and glycine-rich domain. We studied the expression and function of the RNA-binding motif protein 3 (RBM3), a member of this family, in neurons. RBM3 was expressed in multiple brain regions, with the highest levels in cerebellum and olfactory bulb. In dissociated neurons, RBM3 was observed in nuclei and in a heterogeneous population of granules within dendrites. In sucrose gradient assays, RBM3 cofractionated with heavy mRNA granules and multiple components of the translation machinery. Two alternatively spliced RBM3 isoforms that differed by a single arginine residue were identified in neurons; both were post-translationally modified. The variant lacking the spliced arginine exhibited a higher dendritic localization and was the only isoform present in astrocytes. When overexpressed in neuronal cell lines, RBM3 isoforms-enhanced global translation, the formation of active polysomes, and the activation of initiation factors. These data suggest that RBM3 plays a distinctive role in enhancing translation in neurons. PMID- 17403029 TI - Type 2 transglutaminase differentially modulates striatal cell death in the presence of wild type or mutant huntingtin. AB - Huntington's disease (HD), which is caused by an expanded polyglutamine tract in huntingtin (htt), is characterized by extensive loss of striatal neurons. The dysregulation of type 2 transglutaminase (TG2) has been proposed to contribute to the pathogenesis in HD as TG2 is up-regulated in HD brain and knocking out TG2 in mouse models of HD ameliorates the disease process. To understand the role of TG2 in the pathogenesis of HD, immortalized striatal cells established from mice in which mutant htt with a polyglutamine stretch of 111 Gln had been knocked-in and wild type (WT) littermates, were stably transfected with human TG2 in a tetracycline inducible vector. Overexpression of TG2 in the WT striatal cells resulted in significantly greater cell death under basal conditions as well as in response to thapsigargin treatment, which causes increased intracellular calcium concentrations. Furthermore, in WT striatal cells TG2 overexpression potentiated mitochondrial membrane depolarization, intracellular reactive oxygen species production, and apoptotic cell death in response to thapsigargin. In contrast, in mutant striatal cells, TG2 overexpression did not increase cell death, nor did it potentiate thapsigargin-induced mitochondrial membrane depolarization or intracellular reactive oxygen species production. Instead, TG2 overexpression in mutant striatal cells attenuated the thapsigargin-activated apoptosis. When in situ transglutaminase activity was quantitatively analyzed in these cell lines, we found that in response to thapsigargin treatment TG2 was activated in WT, but not mutant striatal cells. These data suggest that mutant htt alters the activation of TG2 in response to certain stimuli and therefore differentially modulates how TG2 contributes to cell death processes. PMID- 17403030 TI - Glutamate release from activated microglia requires the oxidative burst and lipid peroxidation. AB - When activated by proinflammatory stimuli, microglia release substantial levels of glutamate, and mounting evidence suggests this contributes to neuronal damage during neuroinflammation. Prior studies indicated a role for the Xc exchange system, an amino acid transporter that antiports glutamate for cystine. Because cystine is used for synthesis of glutathione (GSH) synthesis, we hypothesized that glutamate release is an indirect consequence of GSH depletion by the respiratory burst, which produces superoxide from NADPH oxidase. Microglial glutamate release triggered by lipopolysaccharide was blocked by diphenylene iodonium chloride and apocynin, inhibitors of NADPH oxidase. This glutamate release was also blocked by vitamin E and elicited by lipid peroxidation products 4-hydroxynonenal and acrolein, suggesting that lipid peroxidation makes crucial demands on GSH. Although NADPH oxidase inhibitors also suppressed nitrite accumulation, vitamin E did not; moreover, glutamate release was largely unaffected by nitric oxide donors, inhibitors of nitric oxide synthase, or changes in gene expression. These findings indicate that a considerable degree of the neurodegenerative consequences of neuroinflammation may result from conversion of oxidative stress to excitotoxic stress. This phenomenon entails a biochemical chain of events initiated by a programmed oxidative stress and resultant mass-action amino acid transport. Indeed, some of the neuroprotective effects of antioxidants may be due to interference with these events rather than direct protection against neuronal oxidation. PMID- 17403031 TI - Hyaluronan-CD44 interaction stimulates Rac1 signaling and PKN gamma kinase activation leading to cytoskeleton function and cell migration in astrocytes. AB - Both hyaluronan [HA, the major glycosaminoglycans in the extracellular matrix (ECM)] and CD44 (a primary HA receptor) are associated with astrocyte activation and tissue repair following central nervous system (CNS) injury. In this study we investigated the question of whether HA-CD44 interaction influences astrocyte signaling and migration. Our data indicated that HA binding to the cultured astrocytes stimulated Rac1 signaling and cytoskeleton-mediated migration. To determine the cellular and molecular basis of these events, we focused on PKN gamma, a Rac1-activated serine/threonine kinase in astrocytes. We determined that HA binding to astrocytes stimulated Rac1-dependent PKN gamma kinase activity which, in turn, up-regulated the phosphorylation of the cytoskeletal protein, cortactin, and attenuated the ability of cortactin to cross-link F-actin. Further analyses indicated that the N-terminal antiparallel coiled-coil (ACC) domains of PKN gamma interacted with Rac1, and transfection of astrocytes with PKN gamma ACCcDNA inhibited PKN gamma activity. Over-expression of the PKN gamma-ACC domain also functions as a dominant-negative mutant to block HA/CD44-mediated PKN gamma activation of cortactin and astrocyte migration. Taken together, these findings strongly suggest that hyaluronan/CD44 interaction with Rac1-PKN gamma plays a pivotal role in cytoskeleton activation and astrocyte migration. These newly discovered HA/CD44-induced astrocyte function may provide important insight into novel therapeutic treatments for tissue repair following CNS injury. PMID- 17403032 TI - Heat-shock protein 105 interacts with and suppresses aggregation of mutant Cu/Zn superoxide dismutase: clues to a possible strategy for treating ALS. AB - A dominant mutation in the gene for copper-zinc superoxide dismutase (SOD1) is the most frequent cause of the inherited form of amyotrophic lateral sclerosis. Mutant SOD1 provokes progressive degeneration of motor neurons by an unidentified acquired toxicity. Exploiting both affinity purification and mass spectrometry, we identified a novel interaction between heat-shock protein 105 (Hsp105) and mutant SOD1. We detected this interaction both in spinal cord extracts of mutant SOD1(G93A) transgenic mice and in cultured neuroblastoma cells. Expression of Hsp105, which is found in mouse motor neurons, was depressed in the spinal cords of SOD1(G93A) mice as disease progressed, while levels of expression of two other heat-shock proteins, Hsp70 and Hsp27, were elevated. Moreover, Hsp105 suppressed the formation of mutant SOD1-containing aggregates in cultured cells. These results suggest that techniques that raise levels of Hsp105 might be promising tools for alleviation of the mutant SOD1 toxicity. PMID- 17403033 TI - Toll-like receptor (TLR)-2 and TLR-4 regulate inflammation, gliosis, and myelin sparing after spinal cord injury. AB - Activation of macrophages via toll-like receptors (TLRs) is important for inflammation and host defense against pathogens. Recent data suggest that non pathogenic molecules released by trauma also can trigger inflammation via TLR2 and TLR4. Here, we tested whether TLRs are regulated after sterile spinal cord injury (SCI) and examined their effects on functional and anatomical recovery. We show that mRNA for TLR1, 2, 4, 5, and 7 are increased after SCI as are molecules associated with TLR signaling (e.g. MyD88, NFkappaB). The significance of in vivo TLR2 and TLR4 signaling was evident in SCI TLR4 mutant (C3H/HeJ) and TLR2 knockout (TLR2-/-) mice. In C3H/HeJ mice, sustained locomotor deficits were observed relative to SCI wild-type control mice and were associated with increased demyelination, astrogliosis, and macrophage activation. These changes were preceded by reduced intraspinal expression of interleukin-1beta mRNA. In TLR2-/- mice, locomotor recovery also was impaired relative to SCI wild-type controls and novel patterns of myelin pathology existed within ventromedial white matter--an area important for overground locomotion. Together, these data suggest that in the absence of pathogens, TLR2 and TLR4 are important for coordinating post-injury sequelae and perhaps in regulating inflammation and gliosis after SCI. PMID- 17403034 TI - Effects of the monomeric, oligomeric, and fibrillar Abeta42 peptides on the proliferation and differentiation of adult neural stem cells from subventricular zone. AB - The incidence of amyloid plaques, composed mainly of beta-amyloid peptides (Abeta), does not correlate well with the severity of neurodegeneration in patients with Alzheimer's disease (AD). The effects of Abeta(42) on neurons or neural stem cells (NSCs) in terms of the aggregated form remain controversial. We prepared three forms of oligomeric, fibrillar, and monomeric Abeta(42) peptides and investigated their effects on the proliferation and neural differentiation of adult NSCs, according to the degree of aggregation or concentration. A low micromolar concentration (1 micromol/L) of oligomeric Abeta(42) increased the proliferation of adult NSCs remarkably in a neurosphere assay. It also enhanced the neuronal differentiation of adult NSCs and their ability to migrate. These results provide us with valuable information regarding the effects of Abeta(42) on NSCs in the brains of patients with AD. PMID- 17403035 TI - Ketone bodies are protective against oxidative stress in neocortical neurons. AB - Ketone bodies (KB) have been shown to prevent neurodegeneration in models of Parkinson's and Alzheimer's diseases, but the mechanisms underlying these effects remain unclear. One possibility is that KB may exert antioxidant activity. In the current study, we explored the effects of KB on rat neocortical neurons exposed to hydrogen peroxide (H(2)O(2)) or diamide - a thiol oxidant and activator of mitochondrial permeability transition (mPT). We found that: (i) KB completely blocked large inward currents induced by either H(2)O(2) or diamide; (ii) KB significantly decreased the number of propidium iodide-labeled cells in neocortical slices after exposure to H(2)O(2) or diamide; (iii) KB significantly decreased reactive oxygen species (ROS) levels in dissociated neurons and in isolated neocortical mitochondria; (iv) the electrophysiological effects of KB in neurons exposed to H(2)O(2) or diamide were mimicked by bongkrekic acid and cyclosporin A, known inhibitors of mPT, as well as by catalase and DL - dithiothreitol, known antioxidants; (v) diamide alone did not significantly alter basal ROS levels in neurons, supporting previous studies indicating that diamide induced neuronal injury may be mediated by mPT opening; and (vi) KB significantly increased the threshold for calcium-induced mPT in isolated mitochondria. Taken together, our data suggest that KB may prevent mPT and oxidative injury in neocortical neurons, most likely by decreasing mitochondrial ROS production. PMID- 17403036 TI - Molecular dynamics of the DNA-binding domain of the papillomavirus E2 transcriptional regulator uncover differential properties for DNA target accommodation. AB - Papillomaviruses are small DNA tumor viruses that infect mammalian hosts, with consequences from benign to cancerous lesions. The Early protein 2 is the master regulator for the virus life cycle, participating in gene transcription, DNA replication, and viral episome migration. All of these functions rely on primary target recognition by its dimeric DNA-binding domain. In this work, we performed molecular dynamics simulations in order to gain insights into the structural dynamics of the DNA-binding domains of two prototypic strains, human papillomavirus strain 16 and the bovine papillomavirus strain 1. The simulations underline different dynamic features in the two proteins. The human papillomavirus strain 16 domain displays a higher flexibility of the beta2-beta3 connecting loop in comparison with the bovine papillomavirus strain 1 domain, with a consequent effect on the DNA-binding helices, and thus on the modulation of DNA recognition. A compact beta-barrel is found in human papillomavirus strain 16, whereas the bovine papillomavirus strain 1 protein is characterized by a loose beta-barrel with a large number of cavities filled by water, which provides great flexibility. The rigidity of the human papillomavirus strain 16 beta-barrel prevents protein deformation, and, as a consequence, deformable spacers are the preferred targets in complex formation. In contrast, in bovine papillomavirus strain 1, a more deformable beta-barrel confers greater adaptability to the protein, allowing the binding of less flexible DNA regions. The flexibility data are confirmed by the experimental NMR S2 values, which are reproduced well by calculation. This feature may provide the protein with an ability to discriminate between spacer sequences. Clearly, the deformability required for the formation of the Early protein 2 C-terminal DNA-binding domain-DNA complexes of various types is based not only on the rigidity of the base sequences in the DNA spacers, but also on the intrinsic deformability properties of each domain. PMID- 17403037 TI - The crystal structure of phenylpyruvate decarboxylase from Azospirillum brasilense at 1.5 A resolution. Implications for its catalytic and regulatory mechanism. AB - Phenylpyruvate decarboxylase (PPDC) of Azospirillum brasilense, involved in the biosynthesis of the plant hormone indole-3-acetic acid and the antimicrobial compound phenylacetic acid, is a thiamine diphosphate-dependent enzyme that catalyses the nonoxidative decarboxylation of indole- and phenylpyruvate. Analogous to yeast pyruvate decarboxylases, PPDC is subject to allosteric substrate activation, showing sigmoidal v versus [S] plots. The present paper reports the crystal structure of this enzyme determined at 1.5 A resolution. The subunit architecture of PPDC is characteristic for other members of the pyruvate oxidase family, with each subunit consisting of three domains with an open alpha/beta topology. An active site loop, bearing the catalytic residues His112 and His113, could not be modelled due to flexibility. The biological tetramer is best described as an asymmetric dimer of dimers. A cysteine residue that has been suggested as the site for regulatory substrate binding in yeast pyruvate decarboxylase is not conserved, requiring a different mechanism for allosteric substrate activation in PPDC. Only minor changes occur in the interactions with the cofactors, thiamine diphosphate and Mg2+, compared to pyruvate decarboxylase. A greater diversity is observed in the substrate binding pocket accounting for the difference in substrate specificity. Moreover, a catalytically important glutamate residue conserved in nearly all decarboxylases is replaced by a leucine in PPDC. The consequences of these differences in terms of the catalytic and regulatory mechanism of PPDC are discussed. PMID- 17403038 TI - Coordination of three and four Cu(I) to the alpha- and beta-domain of vertebrate Zn-metallothionein-1, respectively, induces significant structural changes. AB - Vertebrate metallothioneins are found to contain Zn(II) and variable amounts of Cu(I), in vivo, and are believed to be important for d10-metal control. To date, structural information is available for the Zn(II) and Cd(II) forms, but not for the Cu(I) or mixed metal forms. Cu(I) binding to metallothionein-1 has been investigated by circular dichroism, luminescence and 1H NMR using two synthetic fragments representing the alpha- and the beta-domain. The 1H NMR data and thus the structures of Zn4alpha metallothionein (MT)-1 and Zn3betaMT-1 were essentially the same as those already published for the corresponding domains of native Cd7MT-1. Cu(I) titration of the Zn(II)-reconstituted domains provided clear evidence of stable polypeptide folds of the three Cu(I)-containing alpha- and the four Cu(I)-containing beta-domains. The solution structures of these two species are grossly different from the structures of the starting Zn(II) complexes. Further addition of Cu(I) to the two single domains led to the loss of defined domain structures. Upon mixing of the separately prepared aqueous three and four Cu(I) loaded alpha- and beta-domains, no interaction was seen between the two species. There was neither any indication for a net transfer of Cu(I) between the two domains nor for the formation of one large single Cu(I) cluster involving both domains. PMID- 17403039 TI - The triggers or precipitants of the acute migraine attack. AB - The aim of this study was to evaluate and define the triggers of the acute migraine attack. Patients rated triggers on a 0-3 scale for the average headache. Demographics, prodrome, aura, headache characteristics, postdrome, medication responsiveness, acute and chronic disability, sleep characteristics and social and personal characteristics were also recorded. One thousand two hundred and seven International Classification of Headache Disorders-2 (1.1-1.2, and 1.5.1) patients were evaluated, of whom 75.9% reported triggers (40.4% infrequently, 26.7% frequently and 8.8% very frequently). The trigger frequencies were stress (79.7%), hormones in women (65.1%), not eating (57.3%), weather (53.2%), sleep disturbance (49.8%), perfume or odour (43.7%), neck pain (38.4%), light(s) (38.1%), alcohol (37.8%), smoke (35.7%), sleeping late (32.0%), heat (30.3%), food (26.9%), exercise (22.1%) and sexual activity (5.2%). Triggers were more likely to be associated with a more florid acute migraine attack. Differences were seen between women and men, aura and no aura, episodic and chronic migraine, and between migraine and probable migraine. PMID- 17403040 TI - Apical plug technique using mineral trioxide aggregate: results from a case series. AB - AIM: To examine the clinical and radiographic appearance of teeth that suffered premature interruption of root development and were treated by an mineral trioxide aggregate (MTA) apical plug technique. SUMMARY: Eleven teeth with immature root apices in 11 patients were treated nonsurgically by the manual application of MTA in the apical portion of the root canal under microscopic vision. Follow-up evaluations were performed at 1-2 years after treatment. KEY LEARNING POINTS: Mineral trioxide aggregate appears to be a valid material to obtain periradicular healing in teeth with open apices and necrotic pulps. Ten out of 11 cases were associated with periradicular health at follow-up evaluation. PMID- 17403041 TI - Cytotoxicity analysis of EDTA and citric acid applied on murine resident macrophages culture. AB - AIM: To assess the ex vivo cytotoxicity of EDTA and citric acid solutions on macrophages. METHODOLOGY: The cytotoxicity of 17% EDTA and 15% citric acid was evaluated on murine macrophage cultures using MTT-Tetrazolium method [3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide]. A total of 5 x 10(5) cells were plated in medium culture with 17% EDTA or 15% citric acid. Fresh medium was used as a control. Toxicity values were analysed statistically by anova and Tukey's test (P<0.05) at short (0, 6, 12, 24 h) and medium periods (1, 3, 5, 7 days), using ELISA absorbance. RESULTS: On the short term, both EDTA (0.253 nm) and citric acid (0.260 nm) exhibited cytotoxic effects on macrophage cultures (P<0.05). On the medium term, statistical differences were observed (P<0.05) between the groups. EDTA (0.158 nm) and citric acid (0.219 nm) were cytotoxic when compared with the control group; EDTA-reduced macrophage viability significantly more than citric acid (P<0.05). CONCLUSIONS: Both EDTA and citric acid had effects on macrophages cells ex vivo, but citric acid was less toxic in periods from 1 to 7 days of use. PMID- 17403042 TI - Caspase-8- and JNK-dependent AP-1 activation is required for Fas ligand-induced IL-8 production. AB - Despite a dogma that apoptosis does not induce inflammation, Fas ligand (FasL), a well-known death factor, possesses pro-inflammatory activity. For example, FasL induces nuclear factor kappaB (NF-kappaB) activity and interleukin 8 (IL-8) production by engagement of Fas in human cells. Here, we found that a dominant negative mutant of c-Jun, a component of the activator protein-1 (AP-1) transcription factor, inhibits FasL-induced AP-1 activity and IL-8 production in HEK293 cells. Selective inhibition of AP-1 did not affect NF-kappaB activation and vice versa, indicating that their activations were not sequential events. The FasL-induced AP-1 activation could be inhibited by deleting or introducing the lymphoproliferation (lpr)-type point mutation into the Fas death domain (DD), knocking down the Fas-associated DD protein (FADD), abrogating caspase-8 expression with small interfering RNAs, or using inhibitors for pan-caspase and caspase-8 but not caspase-1 or caspase-3. Furthermore, wildtype, but not a catalytically inactive mutant, of caspase-8 reconstituted the FasL-induced AP-1 activation in caspase-8-deficient cells. Fas ligand induced the phosphorylation of two of the three major mitogen-activated protein kinases (MAPKs): extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) but not p38 MAPK. Unexpectedly, an inhibitor for JNK but not for MAPK/ERK kinase inhibited the FasL-induced AP-1 activation and IL-8 production. These results demonstrate that FasL-induced AP-1 activation is required for optimal IL-8 production, and this process is mediated by FADD, caspase-8, and JNK. PMID- 17403043 TI - C-->G base mutations in the CArG box of c-fos serum response element alter its bending flexibility. Consequences for core-SRF recognition. AB - By binding to the CArG box sequence, the serum response factor (SRF) activates several muscle-specific genes, as well as genes that respond to mitogens. The core domain of the SRF (core-SRF) binds as a dimer to the CArG box C-5C-4A-3T-2A 1T+1T+2A+3G+4G+5 of the c-fos serum response element (SREfos). However, previous studies using 20-mer DNAs have shown that the binding stoichiometry of core-SRF is significantly altered by mutations C-5-->G (SREGfos) and C-5C-4-->GG (SREGGfos) of the CArG box [A Huet, A Parlakian, M-C Arnaud, J-M Glandieres, P Valat, S Fermandjian, D Paulin, B Alpert & C Zentz (2005) FEBS J272, 3105-3119]. To understand these effects, we carried out a comparative analysis of the three 20-mer DNAs SREfos, SREGfos and SREGGfos in aqueous solution. Their CD spectra were of the B-DNA type with small differences generated by variations in the mutual arrangement of the base pairs. Analysis by singular value decomposition of a set of Raman spectra recorded as a function of temperature, revealed a premelting transition associated with a conformational shift in the DNA double helices from a bent to a linear form. Time-resolved fluorescence anisotropy shows that the fluorescein reporter linked to the oligonucleotide 5'-ends experiences twisting motions of the double helices related to the interconversion between bent and linear conformers. The three SREs present various bent populations submitted, however, to particular internal dynamics, decisive for the mutual adjustment of binding partners and therefore specific complex formation. PMID- 17403044 TI - Equilibrium distribution of skeletal actin-tropomyosin-troponin states, determined by pyrene-tropomyosin fluorescence. AB - Actin-tropomyosin-troponin has three structural states, but the functional properties of regulation can be explained with models having two functional states. As a step towards assigning functional properties to all the structural states, we examined fluorescent probes that monitor changes in troponin and tropomyosin. Tropomyosin labeled with pyrene-iodoacetamide is thought to reflect the transition to the most active state, whereas N-((2-iodoacetoxy)ethyl)-N methyl)-amino-7-nitrobenz-2-oxa-1,3-diazole-labeled troponin I is thought to monitor the transition to any state other than the inactive state. The fraction of actin in an active state determined from pyrene excimer fluorescence agreed with that calculated from light-scattering measurements of myosin subfragment 1 (S1)-ADP to regulated actin in both the presence and absence of Ca2+ over a range of ionic strength conditions. The only exceptions were conditions where the binding of S1-ADP to actin was too strong to measure accurately. Pyrene tropomyosin excimer fluorescence was Ca2+ dependent and so reflected the change in population caused by both Ca2+ binding and S1-ADP binding. Pyrene labeling of tropomyosin did not cause a large perturbation of the transition among states of regulated actin. Using pyrene-tropomyosin fluorescence we were able to extend the ionic strength dependence of the parameters describing the co-operativity of binding of S1-ADP to actin as low as 0.1 M. The probes on tropomyosin and troponin I had different responses to Ca2+ and S1-ADP binding. These different sensitivities can be explained by an intermediate between the inactive and active states of regulated actin. PMID- 17403045 TI - The hinge region operates as a stability switch in cGMP-dependent protein kinase I alpha. AB - The molecular mechanism of cGMP-dependent protein kinase activation by its allosteric regulator cyclic-3',5'-guanosine monophosphate (cGMP) has been intensely studied. However, the structural as well as thermodynamic changes upon binding of cGMP to type I cGMP-dependent protein kinase are not fully understood. Here we report a cGMP-induced shift of Gibbs free enthalpy (DeltaDeltaGD) of 2.5 kJ.mol-1 as determined from changes in tryptophan fluorescence using urea-induced unfolding for bovine PKG Ialpha. However, this apparent increase in overall stability specifically excluded the N-terminal region of the kinase. Analyses of tryptic cleavage patterns using liquid chromatography-coupled ESI-TOF mass spectrometry and SDS/PAGE revealed that cGMP binding destabilizes the N-terminus at the hinge region, centered around residue 77, while the C-terminus was protected from degradation. Furthermore, two recombinantly expressed mutants: the deletion fragment Delta1-77 and the trypsin resistant mutant Arg77Leu (R77L) revealed that the labile nature of the N-terminus is primarily associated with the hinge region. The R77L mutation not only stabilized the N-terminus but extended a stabilizing effect on the remaining domains of the enzyme as well. These findings support the concept that the hinge region of PKG acts as a stability switch. PMID- 17403046 TI - Microstructural differences between single-species and dual-species biofilms of Streptococcus mutans and Veillonella parvula, before and after exposure to chlorhexidine. AB - Dual-species biofilms of Streptococcus mutans and Veillonella parvula are less susceptible to antimicrobials than single-species biofilms of the same microorganisms. The microstructure of single and dual-species biofilms of S. mutans and/or V. parvula was visualized to find out whether the spatial arrangement of bacteria in biofilms is related to survival strategies against antimicrobials. Biofilms were grown in glass-bottomed 96-well microtiter plates and exposed to chlorhexidine at 48 h. Fluorescent probes were used for staining. The microstructure of biofilms was analyzed by confocal scanning laser microscopy at 48, 96, 192, and 264 h. Spatial arrangement analysis was performed using DAIME software for 48 h biofilms. A decrease in the viability and thickness in all types of biofilms was detected after chlorhexidine treatment in time. In untreated biofilms, clustering was observed. In chlorhexidine-treated single species biofilms, bacteria were dispersed. However, the most prominent clustering was observed in chlorhexidine-treated dual-species biofilm bacteria, which had a higher survival rate compared with chlorhexidine-treated single-species biofilms. Bacteria in dual-species biofilms establish a specific spatial arrangement, forming clusters within distances below 1.2 microm as a survival strategy against antimicrobials while the same bacteria lack this defensive construction in a single-species biofilm. PMID- 17403047 TI - Interaction of an IHF-like protein with the Rhizobium etli nifA promoter. AB - The nifA gene fulfills an essential role in the regulation of nitrogen fixation genes in Rhizobium etli. Transcription analysis of the nifA gene, assessed using promoter deletions, indicated an oxygen-independent expression, threefold higher during symbiosis as compared with free-living conditions. Electrophoretic mobility shift assays using those nifA promoter deletion fragments, which were actively transcribed, demonstrated the specific interaction with R. etli cellular protein(s) resulting in the formation of two DNA-protein complexes. An interacting protein was purified by liquid chromatography on Heparin Sepharose and Mono S columns. The purified 12 kDa R. etli protein cross-reacted with antibodies directed against Escherichia coli integration host factor (IHF). Furthermore, purified E. coli IHF was able to specifically bind to the R. etli nifA promoter region. These results point to an as yet undisclosed function of IHF in the regulation of R. etli nifA expression. PMID- 17403048 TI - Molecular typing of Leptospira spp. based on putative O-antigen polymerase gene (wzy), the benefit over 16S rRNA gene sequence. AB - Molecular typing of leptospiral strains based on variation within putative O antigen polymerase gene (wzy) was determined among reference strains and those isolated from patients. Using the PCR primers designed from the flanking gene of wzy derived from Leptospira interrogans serovar Copenhageni, all L. interrogans serovars as well as human and rodent leptospiral isolates from Thailand could be amplified. The size of PCR product ranged from 1 to 1.5 kb. The limitation of these primer pairs was the inability to amplify those strains whose sequences differ in the region of the primers, these included Leptospira biflexa (serovar Patoc), Leptospira borgpetersenii (serovar Tarassovi) and Leptospira kirschneri (serovar Bim, Bulgarica, Butembo). Notably, amplification was not limited to L. interrogans as demonstrated by the amplification of some strains from L. kirschneri, Leptospira meyeri, Leptospira noguchii, Leptospira santarosai, L. borgpetersenii and Leptospira weilii. The phylogenetic tree of wzy sequence, inferred by posterior probability of the Bayesian, enabled the categorization of leptospiral serovars into seven genetically related group, of which its differentiation power was better than that of the more highly conserved 16S rRNA gene, which is used extensively for genotyping. PMID- 17403049 TI - The structures of core regions from enterobacterial lipopolysaccharides - an update. AB - To the major virulence factors of Gram-negative bacteria belong the lipopolysaccharides (endotoxins), which are very well characterized for their immunological, pharmacological and pathophysiological effects displayed in eucaryotic cells and organisms. In general, these amphiphilic lipopolysaccharides comprise three regions, which can be differentiated by their structures, function, genetics and biosynthesis: lipid A, the core region and a polysaccharide portion, which may be the O-specific polysaccharide, Enterobacterial Common Antigen (ECA) or a capsular polysaccharide. In the past, much emphasis has been laid on the elucidation of the structure-function relation. The lipid A was proven to represent the toxic principle of endotoxic active lipopolysaccharides, however, its toxicity depends not only on its structure but also on that of the core region, which is covalently linked to lipid A. Thus, and since the core region possesses immunogenic properties, complete structural analyses of lipopolysaccharides core regions and of structure function relation are highly important for a better understanding of lipopolysaccharides action. To date, quite a number of core structures from lipopolysaccharides of various Gram-negative bacteria have been published and summarized in several overviews. This short review adds to this knowledge those structures of enterobacterial lipopolysaccharides that were published between January 2002 and October 2006. PMID- 17403050 TI - Characterization of tccP2 carried by atypical enteropathogenic Escherichia coli. AB - Atypical enteropathogenic Escherichia coli (EPEC) comprise an important group of paediatric pathogens. Atypical EPEC have reservoirs in farm and domestic animals where they can be either commensal or pathogenic; serogroup O26 is dominant in humans and animals. Central to intestinal colonization by EPEC is the translocation of the type III secretion system effector Tir into enterocytes, which following phosphorylation (Tir-Yp) recruits Nck to activate the N-WASP actin signalling cascade. The authors have recently shown that typical EPEC strains, belonging to the EPEC-2 lineage, carry a tir gene encoding Tir-Yp and can also use the alternative TccP2 actin-signalling cascade. The aim of this study was to determine if tccP2 is found in atypical EPEC isolated from human and farm animals. tccP2 was found at a frequency of 41% in non-O26 EPEC isolates and in 82.3% of the O26 strains. TccP2 of human and animal strains show high level of sequence identity. It is shown that most strains carry a tir gene encoding Tir Yp. In addition the authors identified two new variants of tir genes in EPEC O104:H12 and NT:H19 strains. PMID- 17403051 TI - Enhanced induction of cytochromes P450alk that oxidize methyl-ends of n-alkanes and fatty acids in the long-chain dicarboxylic acid-hyperproducing mutant of Candida maltosa. AB - In the long-chain dicarboxylic acids (DCA)-hyperproducing mutant Candida maltosa strains, methyl-ends of n-alkanes and fatty acids are hydroxylated by n-alkane inducible cytochromes P450 (P450alk), presumably as an essential step in DCA production. A significantly higher production of P450alks was observed in response to n-alkane in the DCA-hyperproducing mutant strain M2030 than in the wild-type strain 1098. Northern analysis demonstrated that n-tetradecane induction levels of mRNAs of all four ALK genes encoding major P450alk isoforms involved in n-alkane assimilation were significantly higher in the DCA hyperproducing mutant than in the wild-type strain. Among these four ALK genes, enhancement of the transcriptional induction level of ALK5, which prefers fatty acids as substrates, was prominent in the mutant. In agreement with Northern analysis, promoters of ALK genes, especially that of ALK5, more strongly responded to n-alkanes in the DCA-hyperproducing mutant than in the wild-type strain. These results suggest that the transcriptional control of ALK genes in the DCA-hyperproducing mutant strains was altered preferably to accelerate DCA production. PMID- 17403052 TI - Effect of the leptin c.73T>C mutation on carcass traits in beef cattle. PMID- 17403053 TI - Construction of a river buffalo (Bubalus bubalis) whole-genome radiation hybrid panel and preliminary RH mapping of chromosomes 3 and 10. AB - The buffalo (Bubalus bubalis) is a source of milk and meat, and also serves as a draft animal. In this study, a 5000-rad whole-genome radiation hybrid (RH) panel for river buffalo was constructed and used to build preliminary RH maps for BBU3 and BBU10 chromosomes. The preliminary maps contain 66 markers, including coding genes, cattle expressed sequence tags (ESTs) and microsatellite loci. The RH maps presented here are the starting point for mapping additional loci that will allow detailed comparative maps between buffalo, cattle and other species whose genomes may be mapped in the future. A large quantity of DNA has been prepared from the cell lines forming the river buffalo RH panel and will be made publicly available to the international community both for the study of chromosome evolution and for the improvement of traits important to the role of buffalo in animal agriculture. PMID- 17403054 TI - Additional support for an association between OLR1 and milk fat traits in cattle. AB - The bovine oxidized LDL receptor 1 (OLR1) was chosen as a candidate gene for association tests with milk composition traits. Genotyping of 773 Italian Brown Swiss for a SNP at position 8232 in OLR1 (NW_215807:g.8232C>A) revealed a frequency of 0.95 for the g.8232C allele. The University of Wisconsin Holstein resource population was genotyped for the OLR1 NW_215807:g.[7160C>T; 7161A>G; 8232C>A] SNPs, and four haplotypes were inferred based on the genotypes of sires and their daughters. Oxidized LDL receptor 1 haplotypes were significantly associated with fat percentage (P = 0.0015). Haplotype [C; A; C] was associated with a significant increase in fat percentage when compared with the other haplotypes. PMID- 17403055 TI - Identification of a diagnostic antibody-binding region on the immunogenic protein EpC1 from Echinococcus granulosus and its application in population screening for cystic echinococcosis. AB - An Echinococcus granulosus cDNA sequence coding for EpC1, a proven serodiagnostic marker for cystic echinococcosis (CE, hydatid disease), has high amino acid sequence identity to a paralogue from Taenia solium, the cause of neurocysticercosis (NCC). To determine diagnostic antibody-binding regions on EpC1 recognized specifically by CE sera, 10 truncated regions (P1-10) of the immunogenic protein were expressed in Escherichia coli and subjected to immunoblotting. One peptide, designated peptide 5 [P5, fused with glutathione-S transferase (GST)] was positively recognized by sera from mice experimentally infected with oncospheres of E. granulosus and sera from surgically confirmed CE patients. Sera from NCC patients did not react with any of the peptides used. There are four amino acid substitutions in P5 compared with the T. solium sequence and these may form part of the epitope inducing CE-specific antibody. Ninety-seven per cent (58 of 60) of sera from confirmed CE patients recognized P5 GST, which was higher than the parent EpC1 fused with GST which reacted with 92% (55 of 60) of the sera. A population screening survey showed that 424 human sera collected from communities in Xinjiang, an area in China endemic for CE, exhibited 4.5% and 3.3% positivity in immunoblotting analysis to EpC1 and P5, respectively; 19.8% of these sera reacted positively against hydatid cyst fluid (HCF) antigen B. Low numbers of surgical CE cases have been reported from this population, suggesting that HCF-based serology lacks specificity and that EpC1 or its contained P5 peptide may prove more accurate for seroepidemiological surveys of CE. PMID- 17403056 TI - Role of the mucosal integrin alpha(E)(CD103)beta(7) in tissue-restricted cytotoxicity. AB - The effectiveness of lung transplantation is marred by the relatively high incidence of rejection. The lung normally contains a large population of lymphocytes in contact with the airway epithelium, a proportion of which expresses the mucosal integrin, alpha(E)(CD103)beta(7). This integrin is not a homing receptor, but is thought to retain lymphocytes at the epithelial surface. Following transplantation, a population of 'tissue-restricted' cytotoxic T cells (CTL) have been identified which have the ability to lyse epithelial cells, but not major histocompatibility complex (MHC)-identical splenic cells. We tested the hypothesis that expression of the mucosal integrin confers the ability of CTL to target and destroy e-cadherin expressing targets. Immunohistochemical and flow cytometric analyses were used to demonstrate the relevance of this model to human lung. Allo-activated CTL were generated in mixed leucocyte reactions and CD103 expression up-regulated by the addition of transforming growth factor (TGF)-beta. The functional effect of CD103 expression was investigated in (51)Cr-release assays using e-cadherin-expressing transfectant targets. Human lung epithelial cells express e-cadherin and one-third of intraepithelial lymphocytes (IEL) expressed CD103. Allo-activated and bronchoalveolar lavage (BAL) lymphocytes express more CD103 than those in blood. Transfection of e-cadherin into murine fibroblasts conferred susceptibility to lysis by alpha(E)beta(7)-expressing CTL which could be blocked by specific monoclonal antibodies to CD103 and e-cadherin. CD103 functions to conjugate CTL effectors to e-cadherin-expressing targets and thereby facilitates cellular cytotoxicity. E-cadherin is expressed prominently by epithelial cells in the lung, enabling CTL to target them for destruction. PMID- 17403057 TI - Changes in circulating lymphocyte subpopulations following administration of the leucocyte function-associated antigen-3 (LFA-3)/IgG1 fusion protein alefacept. AB - Alefacept, a recombinant leucocyte function-associated antigen-3 (LFA-3)/IgG1 fusion protein approved for the treatment of psoriasis, is reported to reduce selectively the numbers of circulating CD4(+) CD45RO(+) and CD8(+) CD45RO(+) T cells, while sparing the naive cells. The purpose of the present study was to elucidate further the effect of alefacept on various circulating lymphocyte subsets. Sixteen patients, 12 with chronic plaque psoriasis and four with pustular psoriasis, received alefacept 7.5 mg once weekly for 12 weeks. Blood samples collected at study entry and after 12 weeks of treatment were analysed by four-colour flow cytometry. There were statistically significant reductions in the total number of conventional memory (CD45RA(-) CD27(+)) and effector (CD45RA( ) CD27(-) or CD45RA(+) CD27(-)) T cells, including CD4(+) and CD8(+) T cells expressing CD161 and CD8(+) T cells expressing cutaneous lymphocyte-associated antigen (CLA). Natural killer (NK) T cells were also reduced significantly, while no statistically significant changes were seen in NK cells and CD4(+) CD25(high) cells. The affected subpopulations were all characterized by a high expression of CD2. However, CD4(+) CD25(low), and CD4(+) CLA(+) cells, which also expressed relative high levels of CD2, were not reduced significantly. Our results suggest a heterogeneous effect of alefacept on the circulating memory T cell population, indicating that high expression of CD2 may not, by itself, be sufficient to explain the reduction in cell count for a specific subpopulation. PMID- 17403058 TI - Antibodies to CRMP3-4 associated with limbic encephalitis and thymoma. AB - We present a case with subacute limbic encephalitis (LE) and thymoma. Neither classical onconeural antibodies nor antibodies to voltage gated potassium channels (VGKC) were detected, but the serum was positive for anti-glutamic acid decarboxylase (GAD). The patient serum also stained synaptic boutons of pyramidal cells and nuclei of granule cells of rat hippocampus. The objective of the study was to identify new antibodies associated with LE. Screening a cDNA expression library identified collapsin response mediator protein 3 (CRMP3), a protein involved in neurite outgrowth. The serum also reacted with both CRMP3 and CRMP4 by Western blot. Similar binding pattern of hippocampal granule cells was obtained with the patient serum and rabbit anti-serum against CRMP1-4. The CRMP1 4 antibodies stained neuronal nuclei of a biopsy from the patient's temporal lobe, but CRMP1-4 expression in thymoma could only be detected by immunoblotting. Absorption studies with recombinant GAD failed to abolish the staining of the hippocampal granule cells. Our findings illustrate that CRMP3-4 antibodies can be associated with LE and thymoma. This has previously been associated with CRMP5. PMID- 17403060 TI - Pre-incubation with interleukin-4 mediates a direct protective effect against the loss of pancreatic beta-cell viability induced by proinflammatory cytokines. AB - Loss of pancreatic beta-cells in type I diabetes is associated with an increase in T helper 1 (Th1) proinflammatory cytokines in the islet milieu, with a concomitant reduction in Th2 anti-inflammatory cytokines. In animal models, manoeuvres designed to polarize Th1 responses towards Th2, particularly involving interleukin (IL)-4, have been shown to protect against insulitis and diabetes. The aim of this study was to determine whether IL-4 can exert a direct effect on beta-cell viability. The rat pancreatic beta-cell line, BRIN-BD11, was used. IL 4R mRNA expression was assayed by reverse transcription-polymerase chain reaction and DNA sequencing and protein expression measured using anti-IL-4R antibodies and confocal microscopy. Cells were pretreated in vitro with IL-4, incubated with IL-1beta and interferon (IFN)-gamma and DNA fragmentation and nitrite production analysed by flow cytometry and Griess assay, respectively. Expression of type I (IL-4R alpha and common gamma-chain) and type II (IL-4R alpha, IL-13R alpha-1) IL 4R mRNA transcripts, together with cell surface expression of IL-4R, was demonstrated. Pre-incubation with IL-4 reduced significantly cell death induced by IL-1beta alone or by a combination of IL-1beta and IFN-gamma, although this was not accompanied by a reduced production of nitrite. The protective effect of IL-4 was not seen when all three cytokines were added simultaneously. These results demonstrate, for the first time, expression of IL-4 receptor components on rat pancreatic beta-cells and reveal a direct protective effect on the loss of viability mediated by proinflammatory cytokines when beta-cells are pre-incubated with IL-4. PMID- 17403059 TI - Inhibition of human neutrophil degranulation by transforming growth factor-beta1. AB - Neutrophils enter tissues including the uterus and are found in the endometrium in increased numbers prior to menses. In this environment, they are exposed to transforming growth factor (TGF)-beta1 produced by endometrial stromal and epithelial cells. We observed that incubation of neutrophils in vitro with TGF beta1 at 1 pg/ml significantly reduced their secretion of lactoferrin in response to lipopolysaccharide (LPS). This effect was achieved with as little as 15 min of pretreatment with TGF-beta1. Inhibition of lactoferrin release by TGF-beta1 was observed irrespective of whether neutrophils were stimulated by ligands for Toll like receptor (TLR)-2, TLR-4 or FPR, the G protein-coupled receptor for formylated peptides. Inhibition by TGF-beta1 was negated by SB-431542, a small molecule inhibitor that specifically blocks the kinase activity of the type I TGF beta receptor (ALK5) In contrast to lactoferrin release, another important neutrophil function, interleukin (IL)-8 driven chemotaxis, was not affected by TGF-beta1 at 1 pg/ml or 100 pg/ml. We conclude that in tissues of the female reproductive tract, TGF-beta1 inhibition of neutrophil degranulation may prevent these cells from initiating an inflammatory response or releasing degradative enzymes that could potentially damage the oocyte or fetus. PMID- 17403061 TI - Matrix proteolytic activity during wound healing: modulation by acute ethanol exposure. AB - BACKGROUND: Clinical studies demonstrate that intoxicated patients exhibit an increased incidence of wound healing complications. Previous studies in a murine excisional wound model revealed that acute ethanol exposure impairs the wound healing response, causing decreased angiogenesis and a significant reduction in wound collagen content. METHODS: Using the same murine model of excisional wounding, we examined the effect of a single dose of ethanol on the overall collagen content and collagen type I and type III mRNA expression, transforming growth factor-beta (TGF-beta) production, and levels of several components of the extracellular matrix proteolytic cascade. RESULTS: Wounds from ethanol-treated mice exhibited a significant decrease in collagen and in the production of collagen type I mRNA compared with saline controls. Exposure to ethanol also caused significant increase in wound TGF-beta by day 2 after injury (1.69 +/- 0.29 vs 12.34 +/- 3.97 pg/microg protein, p<0.01). In addition, wounds from mice exposed to ethanol had significantly increased levels of active urokinase plasminogen activator at day 7, (205.10 +/- 48.79 vs 642.70 +/- 159.80 pg/microg protein, p<0.001). The level of matrix metalloproteinase-8, a collagen type I proteinase, was 2.2-fold higher in wounds of ethanol-treated mice compared with control at day 7 (p<0.05). CONCLUSIONS: These studies demonstrate that a single dose of ethanol decreases collagen production, increases the production of TGF beta and increases levels of matrix degrading enzymes. This alteration in protease balance may partially explain the impaired wound healing that follows acute alcohol intoxication. PMID- 17403062 TI - Upper and lower limb motor impairments in alcoholism, HIV infection, and their comorbidity. AB - BACKGROUND: Both HIV infection and alcoholism can impair motor abilities involving manual dexterity and postural stability. Given the high prevalence of HIV and alcoholism comorbidity, we examined whether each disease selectively disrupts different components of upper and lower limb motor control and whether these impairments are compounded by disease comorbidity. METHODS: Simple and complex upper (speed and finger dexterity) and lower (static posture) limb functions were tested in 31 men with HIV infection, 27 with alcoholism, 43 comorbid for HIV infection and alcoholism, and 22 normal healthy controls to assess whether comorbid patients would demonstrate greater motor impairment relative to those with a single diagnosis. RESULTS: Individuals with HIV infection and those with alcoholism had impaired upper and lower limb motor function. Disease comorbidity compounded deficits in speeded finger movement. Neither Beck Depression Inventory scores, self-reported peripheral neuropathy, nor HIV medication accounted for group differences. Lower limb motor composite scores with eyes open were correlated with upper limb motor scores in the alcoholism group. CONCLUSIONS: Overall, the observed impairment patterns indicate the presence of upper and lower limb motor impairment in both HIV infection and alcoholism and the relevance of alcoholism in exacerbating impairment in speeded fine finger movement, when it occurs in HIV infection. PMID- 17403063 TI - Extreme prematurity: an alcohol-related birth effect. AB - BACKGROUND: Rates of preterm delivery, a major proximate cause of perinatal morbidity and mortality, have been increasing. Prenatal alcohol exposure has been implicated in preterm delivery, although results have been inconsistent due to inadequate control for confounding factors, insufficient power, unreliable and inaccurate assessment of both exposure and gestational age, and lack of stratification of prematurity into severity levels. The purpose of this study was to examine the relation between maternal alcohol, cocaine and cigarette use during pregnancy, and extreme and mild preterm birth. METHODS: Three thousand one hundred thirty consecutive gravidas were followed prospectively for antenatal substance use and had ultrasound confirmed pregnancy dating. RESULTS: Alcohol and cocaine, but not cigarette use, were associated with increased risk of extreme preterm delivery after control for potential confounders. For every unit increase in alcohol exposure, risk of extreme preterm delivery increased significantly [odds ratio (OR) 34.8]. Furthermore, in women aged 30+, alcohol exposure was associated with mild prematurity. Abstention from alcohol while continuing to use cocaine and tobacco was related to a decrease in extreme prematurity of 41%. CONCLUSIONS: The risk of extreme preterm delivery associated with alcohol use is substantial and similar in magnitude to other well-recognized risks. Increased accuracy in identifying exposure and the use of ultrasound to confirm gestational age dating likely contributed to the findings of the current study. These findings suggest that eliminating pregnancy alcohol use might substantially reduce the risk of preterm delivery. PMID- 17403064 TI - College attendance and its effect on drinking behaviors in a longitudinal study of adolescents. AB - BACKGROUND: While college attendance has been shown to be associated with increased drinking behaviors, there are no studies to our knowledge that have examined whether college attendance moderates genetic influences for drinking. We first tested for changes in alcohol consumption in adolescents who did and did not subsequently attend college, and then tested for variation of the genetic and environmental determinants of drinking in these 2 groups. METHODS: Participants eligible for this study were selected from 2 samples from the National Longitudinal Study of Adolescent Health (Add Health), a national probability sample (n=7,083) and a sample of sibling pairs (n=855 pairs). Participants were assessed for measures of drinking behaviors as adolescents (wave I) and reinterviewed at 1 (wave II) and 6 years (wave III) following the initial survey. Changes in binge drinking and average quantity of alcohol consumed in the past year were estimated among 4 groups (2-year college students, 4-year college students, college withdrawers, noncollege participants) in sequential cohorts which spanned the ages of 13 to 24 across the 3 Add Health waves. Gene by environment interactions were then tested at wave III using biometrical models in the genetically informative pairs. RESULTS: Participants who did not attend college reported more binge drinking and consumed greater quantities of alcohol as adolescents than participants who subsequently attended college. However, the college students not only surpassed their noncollege peers in alcohol use as young adults, but also exhibited a greater genetic influence on quantity of alcohol consumed per drinking episode. CONCLUSIONS: Exposure to a college environment acts as an environmental moderator, supporting the hypothesis that the magnitude of genetic influence on certain aspects of alcohol consumption is greater in environments where drinking behaviors are more likely to be promoted. PMID- 17403065 TI - Drinking level, neighborhood social disorder, and mutual intimate partner violence. AB - BACKGROUND: Intimate partner violence (IPV) remains a significant public health problem. The purpose of this study is to assess the contribution of drinking patterns to risk for mutual IPV among married/cohabiting adults in the general population, and to determine if the association between drinking level and mutual IPV varies by level of neighborhood social disorder. METHODS: The study sample consists of 19,035 non-Hispanic black, Hispanic, and non-Hispanic white married/cohabiting adults who participated in the 2000 National Household Survey on Drug Abuse (NHSDA), and whose responses were available through the NHSDA public use file. Gender-specific multivariate logistic regression models of mutual IPV were developed to assess the association between drinking level and mutual IPV, and to test whether these associations vary by neighborhood social disorder. RESULTS: Compared with men who are abstainers, men who are past-30 day heavy drinkers are at a more than 6-fold increased risk for mutual IPV. Men in less hazardous drinking categories are at a 2- to 3-fold increased risk of mutual IPV. Neighborhood disorder is independently associated with men's risk for mutual IPV (odds ratio=1.61). Except for women in the most hazardous drinking category, neighborhood disorder moderates the association between women's drinking level and risk of mutual IPV such that risk for mutual IPV significantly increases under conditions of high neighborhood disorder, and decreases to insignificant risk under conditions of low neighborhood disorder. Compared with abstainers, women who are past-30 day heavy drinkers are at an approximate 6-fold risk for mutual IPV regardless of level of neighborhood disorder. CONCLUSIONS: Drinking level and neighborhood characteristics should be taken into account when assessing risk for mutual IPV among married/cohabiting men and women in the general population. An environmental approach to IPV prevention and intervention which addresses the neighborhood context in which couples reside may be a promising strategy for reducing IPV occurrence. PMID- 17403066 TI - Impact of hepatitis C on HIV progression in adults with alcohol problems. AB - BACKGROUND: Coinfection of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) is a substantial medical and public health concern due to its increasing prevalence and complex patient management. Alcohol use may worsen HCV related liver disease and interfere with adherence to antiretroviral therapy (ART) and medical care. We therefore studied the association between HCV infection and markers of HIV disease progression in adults with alcohol problems. METHODS: This is a longitudinal study of 396 HIV-infected persons with alcohol problems, 199 (50%) of whom were coinfected with HCV (positive HCV RNA test). CD4 cell counts and HIV RNA levels were assessed at baseline and then every 6 months for up to 42 months. Hepatitis C virus RNA status was determined at study enrollment. We examined the relationship between HCV infection and laboratory markers of HIV progression (CD4 cell count and log10 HIV RNA) by fitting multivariable longitudinal regression models for each outcome. RESULTS: Among subjects who were adherent to ART, the presence of HCV infection was associated with a lower CD4 cell count (adjusted mean difference -46.0 cells/microL, p=0.03). There was no association observed between HCV infection and CD4 cell count among those not adherent to ART or those not taking ART. No significant association was observed between HCV infection and HIV RNA regardless of ART status. CONCLUSIONS: Hepatitis C virus infection has an adverse effect on CD4 cell count in patients with alcohol problems who are adherent to ART. Addressing HCV coinfection among these patients may confer additional immunologic benefit for this patient population. PMID- 17403067 TI - A randomized trial of a DWI intervention program for first offenders: intervention outcomes and interactions with antisocial personality disorder among a primarily American-Indian sample. AB - BACKGROUND: Randomized trial evidence on the effectiveness of incarceration and treatment of first-time driving while intoxicated (DWI) offenders who are primarily American Indian has yet to be reported in the literature on DWI prevention. Further, research has confirmed the association of antisocial personality disorder (ASPD) with problems with alcohol including DWI. METHODS: A randomized clinical trial was conducted, in conjunction with 28 days of incarceration, of a treatment program incorporating motivational interviewing principles for first-time DWI offenders. The sample of 305 offenders including 52 diagnosed as ASPD by the Diagnostic Interview Schedule were assessed before assignment to conditions and at 6, 12, and 24 months after discharge. Self reported frequency of drinking and driving as well as various measures of drinking over the preceding 90 days were available at all assessments for 244 participants. Further, DWI rearrest data for 274 participants were available for analysis. RESULTS: Participants randomized to receive the first offender incarceration and treatment program reported greater reductions in alcohol consumption from baseline levels when compared with participants who were only incarcerated. Antisocial personality disorder participants reported heavier and more frequent drinking but showed significantly greater declines in drinking from intake to posttreatment assessments. Further, the treatment resulted in larger effects relative to the control on ASPD than non-ASPD participants. CONCLUSIONS: Nonconfrontational treatment may significantly enhance outcomes for DWI offenders with ASPD when delivered in an incarcerated setting, and in the present study, such effects were found in a primarily American-Indian sample. PMID- 17403068 TI - Increased drinking during withdrawal from intermittent ethanol exposure is blocked by the CRF receptor antagonist D-Phe-CRF(12-41). AB - BACKGROUND: Studies in rodents have determined that intermittent exposure to alcohol vapor can increase subsequent ethanol self-administration, measured with operant and 2-bottle choice procedures. Two key procedural factors in demonstrating increased alcohol intake are the establishment of stable alcohol self-administration before alcohol vapor exposure and the number of bouts of intermittent vapor exposure. The present studies provide additional behavioral validation and initial pharmacological validation of this withdrawal-associated drinking procedure. METHODS: Studies at 2 different sites (Portland and Scripps) examined the effect of intermittent ethanol vapor exposure (3 cycles of 16 hours of ethanol vapor+8 hours air) on 2-hour limited access ethanol preference drinking in male C57BL/6 mice. Separate studies tested 10 or 15% (v/v) ethanol concentrations, and measured intake during the circadian dark. In one study, before measuring ethanol intake after the second bout of intermittent vapor exposure, mice were tested for handling-induced convulsions (HICs) indicative of physical dependence on ethanol. In a second study, the effect of bilateral infusions of the corticotropin-releasing factor (CRF) receptor antagonist D-Phe CRF(12-41) (0.25 microg/0.5 microL) into the central nucleus of the amygdala (CeA) on ethanol intake was compared in vapor-exposed animals and air controls. RESULTS: Intermittent ethanol vapor exposure significantly increased ethanol intake by 30 to 40%, and the mice had higher blood ethanol concentrations than controls. Intra-amygdala infusions of D-Phe-CRF(12-41) significantly decreased the withdrawal-associated increase in ethanol intake without altering ethanol consumption in controls. Following the second bout of intermittent vapor exposure, mice exhibited an increase in HICs, when compared with their own baseline scores or the air controls. CONCLUSIONS: Intermittent alcohol vapor exposure significantly increased alcohol intake and produced signs of physical dependence. Initial pharmacological studies suggest that manipulation of the CRF system in the CeA can block this increased alcohol intake. PMID- 17403069 TI - Decision making and binge drinking: a longitudinal study. AB - BACKGROUND: Behavioral decision making, as measured by the Iowa Gambling Task (IGT) is found to be diminished in individuals with substance dependence and other types of disinhibitory psychopathology. However, little is known regarding the relation between heavy alcohol use and decision-making skills in young adults. This study therefore investigated whether binge drinking is related to disadvantageous decision making, as measured by the IGT. We also examined the relation between decision making and impulsivity. METHODS: Latent class growth analysis was used to classify college students into 4 groups (each group n=50, 50% male), based on their binge drinking trajectories over a 2-year time period (precollege through second year of college). Participants were 200 college students, divided in 4 subgroups: (1) low binge drinkers, (2) stable moderate binge drinkers, (3) increasing binge drinkers, and (4) stable high binge drinkers. A measure of decision making, the IGT, impulsivity questionnaires, and multiple indicators of heavy alcohol use were included. RESULTS: The stable high binge-drinking group made less advantageous choices on the IGT than the low binge drinking group. Impulsivity was not related to decision-making performance. Decision-making performance did not differ by gender, but deck preferences and decision time patterns did differ; women preferred low frequency, high amount punishments to a greater extent than men. CONCLUSIONS: Although disadvantageous decision making is related to binge-drinking patterns in emerging adulthood, this relation is independent of impulsivity. Additionally, the association appears attributable to those who engage in heavy (binge) drinking at an early age, but not to age of onset of drinking in general. PMID- 17403071 TI - The impact of curative intent surgery on the survival of pancreatic cancer patients: a U.S. Population-based study. AB - OBJECTIVES: Pancreatic cancer is the fourth leading cause of cancer death in the United States. Curative intent surgery (CIS) is presumed to be the only curative option in the treatment of pancreatic cancer. The determinants and the outcomes of surgical resection are not clear at the population level. METHODS: Using data from nine registries of the Surveillance Epidemiology and End Results (SEER) program, we examined the predictors of receiving CIS as well as the survival of patients receiving CIS between 1987 and 2001. The definition of CIS in SEER is accurate and has been previously validated and found to be highly accurate. Cox proportional hazard model was used to examine the effect of potential determinants on survival. RESULTS: We identified 32,348 cases of pancreatic cancer. Of those, 3,545 (10.9%) received CIS. The proportion of patients receiving CIS decreased significantly with age (P < 0.0001), was similar across all racial groups (10.8% in whites, 11.4% in blacks, 11.5% in Asians, and 11.2 % in Hispanics, P= 0.5), was slightly higher in men (11.3%vs 10.5%, P= 0.02), decreased with advanced disease stage (P < 0.0001), progressively increased over time (7.5% in 1987-89, 9.1% in 1990-92, 10.4% in 1993-95, 12.4% in 1996-98, and 13.4% in 1999-2001, P < 0.0001), and differed significantly across different SEER registries (9.7% in San Francisco, 11.8% in Connecticut, 12.5% in Detroit, 11.7% in Hawaii, 9.3% in Iowa, 10.05% in New Mexico, 9.5% in Seattle, 13.2% in Utah, 11.4% in Atlanta, P < 0.0001). In the multivariable logistic regression analysis, more recent time periods, younger age, early disease stage, and geographic location, but not race or gender, were independent predictors of receiving CIS. In the Cox survival analysis, younger age, early disease stage, more recent time period, geographic location, and receipt of CIS were independent predictors of improved survival and black race was an independent predictor of shorter survival. CONCLUSIONS: Early stage disease, female gender, younger age, geographic location, and more recent time periods are predictors of receiving CIS. Early disease stage and receipt of CIS are the strongest predictors of improved survival among patients with pancreatic cancer. Black race is an independent predictor of shorter survival. PMID- 17403072 TI - The effect of age on response to therapy with peginterferon alpha plus ribavirin in a cohort of patients with chronic HCV hepatitis including subjects older than 65 yr. AB - OBJECTIVES: In many industrialized countries HCV infection is characterized by an increasing prevalence during ageing; however, data on the efficacy of treatment among older patients are scarce. This study was set up to evaluate the effect of age on the treatment of chronic HCV hepatitis with peginterferon alpha plus ribavirin. METHODS: We retrospectively reviewed medical records of 153 adult patients with chronic HCV hepatitis treated with combination therapy; 30 of them (19.6%) were 65 years of age or older. RESULTS: In multivariable analysis, age groups >/=40 years had similar odds of achieving sustained virologic response (P= 0.71) and significantly lower odds of sustained response compared with younger patients (odds ratio [OR] 0.16, 95% confidence interval [CI] 0.05-0.59, P= 0.006; OR 0.13, 95% CI 0.03-0.49, P= 0.002; OR 0.21, 95% CI 0.05-0.91, P= 0.037 for patients aged 40-49 years, 50-64 years, and older than 64 years, respectively). The effect of age was present in the 74 patients infected with genotype 1 or 4 (P= 0.04), while among the 79 patients with genotype 2 or 3 sustained virologic response rates were relatively uniform, with no statistically significant differences. CONCLUSIONS: The probability of good response to combination treatment with peginterferon alpha plus ribavirin is decreased for patients aged more than 40 years infected with genotype 1 or 4, but patients aged more than 65 had a similar rate of response to those aged 40-64 years. Combination treatment may be safely extended to elderly patients with no major contraindications. PMID- 17403073 TI - The efficacy and safety of bile Acid binding agents, opioid antagonists, or rifampin in the treatment of cholestasis-associated pruritus. AB - OBJECTIVES: The objective of this review was to evaluate the efficacy and safety of rifampin, opioid antagonists, or bile acid binding agents in the treatment of cholestasis-related pruritus (CAP) from available randomized controlled trial evidence. METHODS: In addition to a comprehensive gray literature search, the Cochrane Library, MEDLINE, EMBASE, PubMed, and Web of Science were searched. Only full-text RCTs in participants (>75% adult) with CAP on at least one of the three medications were included. The primary outcome was change in pruritus score, recorded as a continuous or dichotomous outcome. Two independent reviewers performed trial selection and quality assessment. RESULTS: From 487 citations, 12 RCTs were included. Rifampin (standardized mean difference [SMD]-1.62, 95% CI 3.05 to -0.18) and opioid antagonists (SMD -0.68, 95% CI -1.19 to -0.17) significantly reduced CAP. The two cholestyramine studies were too heterogeneous to pool. Although cholestyramine (P= 0.35) and rifampin (P= 0.96) were not associated with greater side effects compared with placebo, opioid antagonists were (number needed to harm = 2.6, 95% CI 1.4-25). CONCLUSIONS: The available RCTs are small, few in number, and use varying scales for measuring pruritus. Although both opioid antagonists and rifampin demonstrated a reduction in pruritus, there were insufficient data to judge the efficacy of cholestyramine. Opioid antagonists were associated with transient side effects in a significant proportion of patients. A longer well-designed randomized controlled trial is needed to confirm the efficacy of bile acid binding agents and accurately assess adverse events. PMID- 17403074 TI - A randomized, multicenter, placebo-controlled trial of polyethylene glycol laxative for chronic treatment of chronic constipation. AB - OBJECTIVES: Polyethylene glycol (PEG) 3350 (MiraLAX) is currently approved for the short-term treatment of occasional constipation. This study was designed to compare the safety and efficacy of PEG laxative versus placebo over a 6-month treatment period in patients with chronic constipation. METHODS: Study subjects who met defined criteria for chronic constipation were randomized in this double blind, placebo-controlled, parallel, multicenter study to receive PEG laxative as a single daily dose of 17 g or placebo for 6 months. Baseline constipation status was confirmed during a 14-day observation period. As a primary efficacy variable, treatment success was defined as relief of modified ROME criteria for constipation for 50% or more of their treatment weeks. Various secondary measures were assessed. An Interactive Voice Response System (IVRS) recorded daily bowel movement experience and study efficacy and safety information. Laboratory testing at baseline and monthly for the study duration was analyzed for hematology, blood chemistry including amylase, GGT, uric acid, lipids, and urinalysis. RESULTS: A total of 304 patients were enrolled and received treatment at one of 50 centers. Successful treatment according to the primary efficacy variable was seen in 52.0% of PEG and 11% of placebo subjects (P < 0.001). Similar efficacy was seen in a subgroup of 75 elderly subjects. According to the primary efficacy definition (based on individual treatment weeks), 61% of PEG treatment weeks versus 22% of the placebo weeks were successful (P < 0.001). There were no significant differences in laboratory findings or adverse events except for the gastrointestinal category where diarrhea, flatulence, and nausea were the most frequent with PEG although they were not individually statistically significant compared with placebo. Similar results were observed when analyzed for differences due to gender, race, or age. CONCLUSIONS: PEG laxative is safe and effective for use in patients with chronic constipation for 6 months. PMID- 17403075 TI - Insulated-tip knife endoscopic mucosal resection of large colorectal polyps unsuitable for standard polypectomy. AB - OBJECTIVES: Endoscopic mucosal resection (EMR) has been shown to be safe and effective. En bloc resection is often not achieved using conventional EMR. Insulated-tip knife (It-knife) EMR has been recently proposed for early gastric cancer dissection and removal. This study was conducted to evaluate the safety and efficacy in obtaining en bloc resection with It-knife EMR of large colonic lesions not resectable with standard endoscopic techniques. METHODS: A total of 29 patients (19 men, 10 women, mean age 67.5 yr, range 44-88) were included in the study. Lesions were considered not suitable for standard polypectomy because of large diameter (>3 cm), morphology, and/or position. Lesions were located in the rectum (N = 11), sigmoid: (N = 10), descending: (N = 4), transverse: (N = 2), and hepatic flexure (N = 2). After saline injection, circumferential incision and dissection of the lesions were attempted with the aim of achieving en bloc resection. RESULTS: En bloc resection was achieved in only 55.1% of the lesions (16 out of 29 patients). In the remaining cases, resection was completed with a piecemeal technique. The median size of the en bloc resected specimen was 3 x 3.4 cm. Complications occurred in four patients (13.7%). At histopathology, 13 patients had low-grade dysplasia, 15 high-grade dysplasia. One patient had a tumor invading the submucosa and was submitted to surgery. CONCLUSIONS: It-knife EMR is a promising technique for attempting en bloc resection of large colonic polyps. Adequate training and caution are required because it can be associated with a higher complication rate than with other EMR modalities. PMID- 17403076 TI - Effect of a proton pump inhibitor or an H2-receptor antagonist on prevention of bleeding from ulcer after endoscopic submucosal dissection of early gastric cancer: a prospective randomized controlled trial. AB - OBJECTIVES: With conventional methods of endoscopic mucosal resection for early gastric cancer (EGC), proton pump inhibitors (PPIs) and H2-receptor antagonists (H2RAs) have a similar effect on preventing bleeding from artificial ulcers. An objective of this study is to investigate whether a stronger acid suppressant (i.e., PPI) more effectively prevents bleeding after the recent advanced technique of endoscopic submucosal dissection (ESD) for EGC. METHODS: This was a prospective randomized controlled trial performed in a referral cancer center. A total of 143 patients with EGC who underwent ESD were randomly assigned to the treatment groups. They received either rabeprazole 20 mg (PPI group) or cimetidine 800 mg (H2RA group) on the day before ESD and continued for 8 wk. The primary end point was the incidence of bleeding that was defined as hematemesis or melena that required endoscopic hemostasis and decreased the hemoglobin count by more than 2 g/dL. RESULTS: In baseline data, the endoscopists who performed the ESD were significantly different between the groups. Finally, 66 of 73 patients in the PPI group and 64 of 70 in the H2RA group were analyzed. Bleeding occurred in four patients in the PPI group and 11 in the H2RA group (P= 0.057). Multivariate analysis revealed that treatment with the PPI significantly reduced the risk of bleeding: adjusted hazard ratio 0.47, 95% confidence interval 0.22 0.92, P= 0.028. One delayed perforation was experienced in the H2RA group. CONCLUSIONS: PPI therapy more effectively prevented delayed bleeding from the ulcer created after ESD than did H2RA treatment. PMID- 17403077 TI - Endomyocardial biopsy guided by electroanatomic voltage mapping in arrhythmogenic right ventricular cardiomyopathy: a case report. AB - A positive endomyocardial biopsy (EMB) is a major diagnostic criterion for arrhythmogenic right ventricular cardiomyopathy (ARVC). Nevertheless, its sensitivity is low due to the focal nature of the disease. Moreover, myocardial samples are usually taken from the uncommonly involved interventricular septum to minimize the risk of perforation. In this report, we describe a novel bioptical approach for ARVC diagnosis guided by the identification of right ventricle (RV) affected regions by means of electroanatomical voltage mapping. PMID- 17403078 TI - Adverse interaction between a left ventricular assist device and an implantable cardioverter defibrillator. AB - An increasing number of patients have a coexisting implantable cardioverter defibrillator (ICD) and left ventricular assist device (LVAD) to treat ventricular arrhythmias and refractory heart failure, respectively. To date, there have been no published reports of negative interactions between these devices that have impacted appropriate ICD or LVAD function. In this case report, we describe a patient with an LVAD-ICD interaction that necessitated replacement of the implantable defibrillator. PMID- 17403079 TI - Rapid ablation for atrial flutter by targeting maximum voltage-factors associated with short ablation times. AB - BACKGROUND: The maximum voltage-guided (MVG) approach to ablation for atrial flutter targets high-amplitude signals along the cavotricuspid isthmus (CTI). It is based on the observation that the isthmus is often composed of bundles of conducting tissue and the hypothesis that these bundles manifest as high amplitude electrograms, providing targets for selective ablation. We aim to identify patient and procedural factors that correlate with rapid isthmus ablation. METHODS: All patients undergoing CTI ablation at our center from January 2005 to May 2006 were included. Patients were divided into outcome groups relative to the median value for total ablation time. The two groups were compared according to patient and procedural variables, using multivariate regression methods. RESULTS: Seventy-six patients were assessed with mean age 60.2 +/- 10.6 years; 63 (82.9%) were male. Mean ablation time to bidirectional block across the CTI was 6.85 +/- 5.87 min (range 0.68-28.7); median 4.77 min. Seventy-six percent of patients required less than 5 min total ablation time until bidirectional block was achieved. Variables independently associated with a short ablation time were the presence of sinus rhythm at start of ablation (P = 0.0050, odds ratio (OR) 8.03), high mean temperature among all ablations (P = 0.019, OR 17.81), and low variability of mean power among all ablations (P = 0.0048, OR 19.26). CONCLUSIONS: Using the MVG approach to atrial flutter ablation, shorter total ablation times are observed among patients in sinus rhythm at the onset of ablation, with higher mean temperature among ablation lesions, and less variability of power between ablations. PMID- 17403080 TI - Optimal electrogram voltage recording technique for detecting the acute ablative tissue injury in the human right atrium. AB - BACKGROUND: The optimal recording technique of the electrogram voltage for detecting abnormal atrial tissue remains unclear. The aim of this study was to compare the impact of various recording techniques on the electrogram voltage after the delivery of ablation therapy in the human right atrium (RA). MATERIAL AND METHODS: Noncontact mapping was performed in 27 patients with typical atrial flutter (mean age = 63 +/- 16, males = 20). Noncontact unipolar and bipolar electrograms were obtained before and after cavotricuspid isthmus (CTI) linear ablation. All unipolar electrograms were acquired with both wide-band filtering (0.5-300 Hz) and narrow-band filtering (32-300 Hz). The unipolar voltage measurements included both the peak-to-peak voltage and peak-negative voltage (PNV) for both filter settings. RESULTS: A comparison of the electrogram voltage along the ablation line before and after the ablation demonstrated a greater reduction in the unipolar PNV with wide-band filtering (70 +/- 24%) than in any of the other recording modalities (P = 0.03). It was the most sensitive and specific recording technique to predict conduction block (cut-off Value 0.35 mV; sensitivity = 94.4% and specificity = 80%). A comparison of the electrogram voltage between the ablated atrial myocardium and nearby nonablated myocardium showed that the unipolar PNV with the wide-band filtering remained the most sensitive method to detect the acute ablative tissue injury, whereas the peak-to peak bipolar voltage was the most specific method. CONCLUSION: The noncontact unipolar electrogram using the PNV with wide-band filter settings (0.5-300 Hz) provided the most sensitive recording technique for detecting acute ablative tissue injury. PMID- 17403081 TI - About arrhythmogenic triggers and substrates in aging atria: partners in crime for atrial fibrillation development? PMID- 17403082 TI - Dynamic changes of T-wave alternans: does it predict short-term arrhythmia vulnerability? PMID- 17403083 TI - If it is not broken, don't fix it: avoidance of right ventricular pacing in cardiac resynchronization therapy. PMID- 17403085 TI - Feasibility and accuracy of a rapid 'point-of-care' D-dimer test performed with a capillary blood sample. PMID- 17403086 TI - Polymorphism 10034C>T is located in a region regulating polyadenylation of FGG transcripts and influences the fibrinogen gamma'/gammaA mRNA ratio. AB - BACKGROUND: Fibrinogen gamma haplotype 2 (FGG-H2) is associated with reduced fibrinogen gamma' levels and fibrinogen gamma'/total fibrinogen ratios and with an increased deep-venous thrombosis (DVT) risk. Two FGG-H2 tagging single nucleotide polymorphisms (SNPs), 9615C>T and 10034C>T, are located in the region of alternative FGG pre-mRNA processing. 10034C>T is located in a GT-rich downstream sequence element (DSE) that comprises a putative cleavage stimulation factor (CstF) binding site. OBJECTIVES: To investigate the functionality of SNPs 9615C>T and 10034C>T, and the importance of the DSE containing 10034C>T. METHODS: Different minigene constructs containing FGG exon 9, intron 9, exon 10 and the 3' region were transiently transfected into HepG2 cells and quantitative real-time polymerase chain reaction was used to measure relative polyadenylation (pA) signal usage (pA1/pA2 ratio). RESULTS: Compared with the reference construct CC (9615C-10034C; FGG-H1; pA1/pA2 ratio set at 100%), the pA1/pA2 ratio of construct TT (9615T-10034T; FGG-H2) was 1.4-fold decreased (71.5%, P = 0.015). The pA1/pA2 ratio of construct CT (9615C-10034T) was almost 1.2-fold decreased (85.3%, P = 0.001), whereas the pA1/pA2 ratio of construct TC (9615T-10034C) did not differ significantly from the reference construct (101.6%, P = 0.890). Functionality of the putative CstF binding site was confirmed using constructs in which this site was deleted or its sequence altered by point mutations. CONCLUSIONS: SNP 10034C>T is located in a GT-rich DSE involved in regulating the usage of the pA2 signal of FGG, which may represent a CstF binding site. We propose that the 10034C>T change is the functional variation in FGG-H2 that is responsible for the reduction in the fibrinogen gamma'/total fibrinogen ratio and the increased DVT risk. PMID- 17403087 TI - Effect of obesity on outcomes after fondaparinux, enoxaparin, or heparin treatment for acute venous thromboembolism in the Matisse trials. AB - BACKGROUND: Selecting initial anticoagulant dose by patient weight for acute pulmonary embolism and deep vein thrombosis has clinical credibility; however, uncertainty remains regarding how to dose obese patients with newer anticoagulants because outcome data are sparse. OBJECTIVES: To use the Matisse trials' comparison of sc fondaparinux once daily with control heparin therapies (intravenous unfractionated heparin for pulmonary embolism, sc enoxaparin 1 mg/kg b.i.d. for deep vein thrombosis) for initial treatment in order to compare primary outcomes (venous thromboembolism recurrence and major bleeding) in obese patients. PATIENTS AND METHODS: Primary outcomes were compared in subsets composed of patients weighing < or = and > 100 kg and with body mass index (BMI) < 30 and > or = 30 kg/m(2). Medians and ranges for weight and BMI were compared for patients suffering either recurrence or major bleeding. RESULTS: Twenty-two thousand and one patients received fondaparinux and 2217 received enoxaparin or unfractionated heparin. Four hundred and ninety-six patients (11%) weighed > 100 kg and 1216 (28%) had a BMI > or = 30. Treatment groups had similar characteristics. The upper limit in subject weight for recurrence was 166 kg (BMI 58), and for major bleeding 120 kg (BMI 39). The incidences of recurrence and major bleeding were similar for each patient subset of weight and BMI for both fondaparinux and heparin treatment groups. Among patients with a primary outcome, median weights and BMIs were also similar. CONCLUSIONS: The current recommended doses of fondaparinux and heparins for the treatment of venous thromboembolism appear to provide similar protection against recurrence and major bleeding to one another and to obese and non-obese patients. PMID- 17403088 TI - Calibration of the WHO 1st International Standard for blood coagulation factor V in plasma, human (03/116). PMID- 17403089 TI - Bleeding events and other complications during pregnancy and childbirth in women with von Willebrand disease. AB - BACKGROUND: Case reports and case series suggest that women with von Willebrand disease (VWD) are at an increased risk of bleeding complications during pregnancy and delivery. OBJECTIVES: To estimate the incidence of bleeding events and other complications in women with VWD during pregnancy and childbirth. METHODS: The United States Nationwide Inpatient Sample (NIS) from the Healthcare Cost and Utilization Project of the Agency for Healthcare Research and Quality for the years 2000-2003 was queried for all pregnancy-related discharges. Women with a diagnosis of VWD were compared with women without VWD. Data were analyzed based on the NIS sampling design. Logistic regression was used to compute odds ratios with 95% CI. RESULTS: There were 4067 deliveries among women with VWD (1 in 4000 deliveries). Although women with VWD were more likely to experience antepartum bleeding [odds ratio (OR) 10.2, 95% CI: 7.1, 14.6], they were no more likely to experience premature labor, placental abruption, fetal growth restriction or intrauterine fetal demise. Women with VWD were more likely to experience a postpartum hemorrhage (OR, 1.5; 95% CI: 1.1, 2.0), and had a 5-fold increased risk of being transfused (OR, 4.7; 95% CI: 3.2, 7.0). Five of the 4067 women with VWD died, a maternal mortality rate 10 times higher than that for other women. CONCLUSIONS: Although women with VWD do not appear to be at an increased risk of poor fetal outcomes, they are at an increased risk of bleeding events and possibly death during pregnancy and childbirth. PMID- 17403090 TI - Treatment of severe von Willebrand disease with a high-purity von Willebrand factor concentrate (Wilfactin): a prospective study of 50 patients. AB - BACKGROUND AND OBJECTIVES: A plasma-derived von Willebrand factor (VWF) concentrate with low factor VIII (FVIII) content was specifically developed to treat von Willebrand disease (VWD). Efficacy and safety were investigated by merging the results of two comparable protocols conducted prospectively in 5 European and 12 French centers. METHODS AND RESULTS: Fifty patients with clinically severe VWD (72% had VWF ristocetin cofactor activity less than 10 IU dL(-1) and 46% had FVIII < 20 IU dL(-1)) were treated with the concentrate as the only therapy, except for clinical situations requiring a priming dose of FVIII to rapidly correct an intrinsic coagulation defect. A total of 139 spontaneous bleeding episodes were treated; only 53 (38%) needed a concomitant FVIII dose. Outcome was excellent or good in 89% of the episodes. Forty-four patients underwent 108 surgical or invasive procedures. Outcome was excellent or good in 95 scheduled procedures (only VWF was infused) and 13 emergency procedures (a priming FVIII dose was co-administered with the first VWF infusion). There were no thrombotic complications and none of the 18 patients with type 3 VWD developed anti-VWF or anti-FVIII antibodies. CONCLUSIONS: This concentrate safely and effectively provides hemostasis in patients with clinically severe VWD. PMID- 17403091 TI - A comparison of anticoagulation results of patients managed with narrow vs. standard international normalized ratio target ranges. PMID- 17403093 TI - The suppressive effect of sphingosine 1-phosphate on monocyte-endothelium adhesion may be mediated by the rearrangement of the endothelial integrins alpha(5)beta(1) and alpha(v)beta(3). AB - BACKGROUND: Sphingosine 1-phosphate (S1P), known to play important roles in vascular biology, is a bioactive lysophospholipid mediator that maintains endothelial integrity via its cell-surface receptors (S1Ps). In this in vitro study, we aimed to examine the role of S1P in monocyte-endothelium adhesion, which is an important event in the pathophysiology of atherosclerosis. METHODS AND RESULTS: S1P pretreatment of human umbilical vein endothelial cells (ECs), but not U937 cells, effectively suppressed U937-EC adhesion independently from the expression of adhesion molecules, namely ICAM-1, VCAM-1, and E-selectin. This S1P-induced suppressive effect was inhibited by the blockage of S1P(1) and S1P(3) receptors and the specific inhibitors of G(i) protein, Src family proteins, phosphatidylinositol 3-kinase, and Rac1, indicating involvement of these key downstream pathways. Moreover, the RGD peptide and antibodies, which neutralize adhesion via alpha(5)beta(1) and alpha(v)beta(3), effectively inhibited U937-EC adhesion with a degree similar to S1P pretreatment. Both an adhesion assay and flow-cytometric analysis demonstrated that U937 cells adhered through integrins alpha(5)beta(1) and alpha(v)beta(3) expressed on the apical surface of monolayer ECs, and S1P shifted the localization of these integrins from the apical surface to the basal surface. CONCLUSIONS: From the present results, we propose that S1P may contribute to the maintenance of vascular integrity and the regulation of atherogenesis through the rearrangement of endothelial integrins. PMID- 17403094 TI - Local tissue-type plasminogen activator release in patients with ischemic stroke. PMID- 17403095 TI - Enhanced peripheral thrombogenicity after lung inflammation is mediated by platelet-leukocyte activation: role of P-selectin. AB - BACKGROUND: Inhaled ultrafine particles trigger peripheral thrombotic complications. METHODS: We have analyzed the systemic prothrombotic risk following lung inflammation induced by pulmonary carbon nanotubes (CNTs). RESULTS: Intratracheal instillation in Swiss mice of 200 and 400 microg of multiwall ground CNTs triggered substantial lung neutrophil, but not macrophage influx, 24 h later. The detection of circulating platelet-leukocyte conjugates exclusively 6 h after CNT instillation pointed to early but transient activation of circulating platelets. At 24 h, elevated plasma procoagulant microvesicular tissue factor activity was found in CNT-exposed but not in saline-exposed mice. However, at 24 h, both the tail and jugular vein bleeding times were prolonged in CNT-exposed but not in saline-exposed mice, arguing against strong CNT-induced platelet activation at this point. Nevertheless, at 24 h, enhanced peripheral thrombogenicity was detected in CNT-exposed but not in saline-exposed mice, via quantitative photochemically induced carotid artery thrombosis measurements. P selectin neutralization abrogated platelet-leukocyte conjugate formation and microvesicular tissue factor generation, and abolished the CNT-induced thrombogenicity amplification. In contrast, the weak vascular injury-triggered thrombus formation in saline-treated mice was not affected by P-selectin neutralization at 24 h. CONCLUSIONS: The mild CNT-induced lung inflammation translates via rapid but mild and transient activation of platelets into P selectin-mediated systemic inflammation. Leukocyte activation leads to tissue factor release, in turn eliciting inflammation-induced procoagulant activity and an associated prothrombotic risk. PMID- 17403096 TI - Fibrinolysis in a lipid environment: modulation through release of free fatty acids. AB - BACKGROUND: Thrombolysis is conventionally regarded as dissolution of the fibrin matrix of thrombi by plasmin, but the structure of clots in vivo includes additional constituents (proteins, phospholipids) that modulate their solubilization. OBJECTIVE: We examined the presence of free fatty acids in thrombi and their effects on distinct stages of fibrinolysis (plasminogen activation, plasmin activity). METHODS AND RESULTS: Using the fluorescent probe acrylodated intestinal fatty acid-binding protein, variable quantities (up to millimolar concentrations) of free fatty acids were demonstrated in surgically removed human thrombi. Oleic acid at relevant concentrations reversibly inhibits more than 90% of the amidolytic activity of plasmin on a synthetic substrate (Spectrozyme PL), but only partially inhibits its fibrinolytic activity measured using turbidimetry. Chromogenic assays detecting the generated plasmin activity show that plasminogen activation by tissue-type plasminogen activator (t-PA) is completely blocked by oleic acid in the fluid phase, but is accelerated on a fibrin matrix. A recombinant derivative of t-PA (reteplase) develops higher fibrin specificity in the presence of oleic acid, because both the inhibition of plasminogen activation in free solution and its enhancement on fibrin template are stronger than with wild-type t-PA. CONCLUSION: Through the stimulation of plasminogen activation on a fibrin template and the inhibition of plasminogen activators and plasmin in the fluid phase, free fatty acids confine the action of fibrinolytic proteases to the site of clotting, where they partially oppose the thrombolytic barrier function of phospholipids. PMID- 17403097 TI - Microsomal prostaglandin E synthase-1, which is not coupled to a particular cyclooxygenase isoenzyme, is essential for prostaglandin E(2) biosynthesis in vascular smooth muscle cells. AB - BACKGROUND: Prostaglandin (PG) E(2) induces expression of matrix metalloproteinases and angiogenic factors, thereby contributing to plaque instability. OBJECTIVE: To study the influence of cyclooxygenase (COX) and PGE synthase (PGES) isoenzyme expression on PGE(2) and PGI(2) biosynthesis in vascular smooth muscle cells (VSMC) in culture. METHODS: Cells were treated with human recombinant IL-1beta over different periods of time. Expression of PGI synthase, and COX and PGES isoenzymes was determined by real-time reverse transcriptase polymerase chain reaction and immunoblotting. Biosynthesis of prostanoids from exogenous or endogenous substrate was analyzed by high performance liquid chromatography or enzyme-immunoassay after incubation of cells with labeled arachidonic acid or thrombin, respectively. RESULTS: Cytosolic PGES and microsomal PGES (mPGES) -1 and -2 were expressed in VSMC. PGES activity was mainly linked to mPGES-1. IL-1beta induced COX-2 and mPGES-1 with a different time course. VSMC ability to synthesize PGE(2) and PGI(2) fitted mPGES-1 and COX 2 expression, respectively. The ability of VSMC to produce PGI(2) was downregulated by mPGES-1 expression and was restored when mPGES-1 expression was silenced. Results from COX-1 and COX-2 silencing and selective inhibition showed that both COX-1 and COX-2 were involved in the biosynthesis of PGE(2) and their relative contribution depended on the time of incubation with IL-1beta. CONCLUSIONS: mPGES-1 is the main PGES responsible for PGE(2) biosynthesis by VSMC and its induction downregulates VSMC ability to produce PGI(2.) These results support the concept that under inflammatory conditions VSMC could significantly contribute to plaque instability and that mPGES-1 may be a target for therapeutic intervention in patients with cardiovascular risk. PMID- 17403098 TI - Linkage analysis for three coagulation factors clustering on chromosome 13q34: factor VII, factor X and protein Z. PMID- 17403099 TI - High risk for venous thromboembolism in diabetics with hyperosmolar state: comparison with other acute medical illnesses. AB - BACKGROUND: Diabetes mellitus is generally not recognized as an important risk factor for venous thromboembolism (VTE). However, clinical observations and case reports have suggested that patients with diabetes and hyperosmolarity may be at increased risk for VTE. OBJECTIVES: To determine the risk of VTE in patients hospitalized for diabetes with hyperosmolar state compared to patients with other acute medical illnesses. PATIENTS/METHODS: The California Patient Discharge Data Set was used to determine the incidence of first-time VTE in all patients admitted between 1995 and 2000 for diabetes with hyperosmolarity and 11 other acute medical conditions. Proportional hazard modeling was used to adjust for age, race, gender, and prior hospitalization within 3 months. RESULTS: Among 2859 patients with diabetes and hyperosmolarity, 34 (1.2%) developed VTE during the hospitalization and 14 (0.5%) developed VTE within 91 days after discharge. In an adjusted multivariate model comparing the risk of VTE to cases with depression, patients with hyperosmolarity had a significantly higher risk of VTE [hazard ratio (HR) = 16.3; 95% confidence interval (CI): 10-25] comparable to the risk associated with sepsis (HR = 19.3; 95% CI: 13-29) or acute connective tissue disease (HR = 21; 95% CI: 15-31). Compared to uncomplicated diabetes, patients with hyperosmolarity had a significantly higher risk of VTE (HR = 3.0; 95% CI: 2.1-4.5) whereas patients with ketoacidosis were not at higher risk (HR = 1.2; 95% CI: 0.8-1.7). CONCLUSIONS: Patients hospitalized for diabetes with hyperosmolarity are at increased risk for developing VTE both during their inpatient stay and in the 3 months after discharge. Thromboprophylaxis in these patients appears warranted, and extended prophylaxis for after hospital discharge should be studied. PMID- 17403100 TI - A note on permutation tests for variance components in multilevel generalized linear mixed models. AB - In many applications of generalized linear mixed models to multilevel data, it is of interest to test whether a random effects variance component is zero. It is well known that the usual asymptotic chi-square distribution of the likelihood ratio and score statistics under the null does not necessarily hold. In this note we propose a permutation test, based on randomly permuting the indices associated with a given level of the model, that has the correct Type I error rate under the null. Results from a simulation study suggest that it is more powerful than tests based on mixtures of chi-square distributions. The proposed test is illustrated using data on the familial aggregation of sleep disturbance. PMID- 17403101 TI - Combining cross-sectional and prospective data methods to improve transition parameter estimation for characterizing the accumulation of HIV-1 drug resistance mutations. AB - The order and rate of acquisition of HIV drug resistance mutations have been estimated from longitudinal and cross-sectional data using Markov models and branching trees, respectively. This article proposes methods that make use of both longitudinal and cross-sectional data simultaneously by employing link functions between the two parameter sets. Most functions that link the two parameter sets also depend on the time on treatment before the start of the study information that may not be available. Nonetheless, under certain assumptions, some link functions eliminate the dependence on time. Using such functions, the two sources of information can be combined to improve the precision of parameter estimation. The method also accommodates error in the link functions from uncertainty in the assumptions required for the links or other reasons. These methods are applied to data from AIDS Clinical Trial Group protocol 398, a randomized comparison of mono- versus dual-protease inhibitor use in heavily treatment experienced HIV patients. Combining the two sources of information allows detection of differences between rates of transition that are not detectable using prospective data alone. PMID- 17403102 TI - Estimating population size for a continuous time frailty model with covariates in a capture-recapture study. AB - A continuous time frailty capture-recapture model is proposed for estimating population size of a closed population with the use of observed covariates to explain individuals' heterogeneity in presence of a random effect. A conditional likelihood approach is used to derive the estimate of parameters, and the Horvitz Thompson estimator is adopted to estimate the unknown population size. Asymptotic normality of the estimates is obtained. Simulation results and a real example show that the proposed method works satisfactorily. PMID- 17403103 TI - Designs for phase I clinical trials with multiple courses of subjects at different doses. AB - The goal of this article is to provide a new design framework and its corresponding estimation for phase I trials. Existing phase I designs assign each subject to one dose level based on responses from previous subjects. Yet it is possible that subjects with neither toxicity nor efficacy responses can be treated at higher dose levels, and their subsequent responses to higher doses will provide more information. In addition, for some trials, it might be possible to obtain multiple responses (repeated measures) from a subject at different dose levels. In this article, a nonparametric estimation method is developed for such studies. We also explore how the designs of multiple doses per subject can be implemented to improve design efficiency. The gain of efficiency from "single dose per subject" to "multiple doses per subject" is evaluated for several scenarios. Our numerical study shows that using "multiple doses per subject" and the proposed estimation method together increases the efficiency substantially. PMID- 17403104 TI - Fixed and random effects selection in linear and logistic models. AB - We address the problem of selecting which variables should be included in the fixed and random components of logistic mixed effects models for correlated data. A fully Bayesian variable selection is implemented using a stochastic search Gibbs sampler to estimate the exact model-averaged posterior distribution. This approach automatically identifies subsets of predictors having nonzero fixed effect coefficients or nonzero random effects variance, while allowing uncertainty in the model selection process. Default priors are proposed for the variance components and an efficient parameter expansion Gibbs sampler is developed for posterior computation. The approach is illustrated using simulated data and an epidemiologic example. PMID- 17403105 TI - Sharp simultaneous confidence intervals for the means of selected populations with application to microarray data analysis. AB - Simultaneous inference for a large number, N, of parameters is a challenge. In some situations, such as microarray experiments, researchers are only interested in making inference for the K parameters corresponding to the K most extreme estimates. Hence it seems important to construct simultaneous confidence intervals for these K parameters. The naive simultaneous confidence intervals for the K means (applied directly without taking into account the selection) have low coverage probabilities. We take an empirical Bayes approach (or an approach based on the random effect model) to construct simultaneous confidence intervals with good coverage probabilities. For N = 10,000 and K = 100, typical for microarray data, our confidence intervals could be 77% shorter than the naive K-dimensional simultaneous intervals. PMID- 17403106 TI - Bayesian adaptive regression splines for hierarchical data. AB - This article considers methodology for hierarchical functional data analysis, motivated by studies of reproductive hormone profiles in the menstrual cycle. Current methods standardize the cycle lengths and ignore the timing of ovulation within the cycle, both of which are biologically informative. Methods are needed that avoid standardization, while flexibly incorporating information on covariates and the timing of reference events, such as ovulation and onset of menses. In addition, it is necessary to account for within-woman dependency when data are collected for multiple cycles. We propose an approach based on a hierarchical generalization of Bayesian multivariate adaptive regression splines. Our formulation allows for an unknown set of basis functions characterizing the population-averaged and woman-specific trajectories in relation to covariates. A reversible jump Markov chain Monte Carlo algorithm is developed for posterior computation. Applying the methods to data from the North Carolina Early Pregnancy Study, we investigate differences in urinary progesterone profiles between conception and nonconception cycles. PMID- 17403107 TI - Recombinant adeno-associated virus vector for gene transfer to the transplanted rat heart. AB - Efficient durable viral vector transduction of the transplanted heart remains elusive. This study assesses the potential of recombinant adeno-associated virus (rAAV) mediated gene delivery to the transplanted rat heart. rAAV serotype 1, 2 and 5 vectors encoding the green fluorescent protein (GFP) gene (1 x 10(11) viral particles/ml) were diluted in cold University of Wisconsin solution and circulated through the coronary vasculature of the donor organs for 30 min before syngeneic rat heterotopic heart transplantation was performed. Study 1: animals (n = 5 each serotype) were killed at 21 days post-transplant to evaluate the efficiency of GFP transduction using RT-PCR and expression by fluorescence microscopy. Study 2: using rAAV-1, animals (n = 5 each group) were killed at 7, 21 and 84 days to evaluate the durability of GFP expression. The maximum cardiac GFP expression at 21 days was observed in rAAV-1. GFP expression by rAAV-1 was detectable at 7 days, improved at 21 days, and was still evident at 84 days. This study demonstrates cardiac rAAV gene transduction with a cold perfusion preservation system of the donor heart. These data show that AAV-1 is superior to AAV-2 and AAV-5 for this purpose and that durable expression is achievable. PMID- 17403108 TI - Heart transplantation in insulin-treated diabetic mellitus patients with diabetes related complications. AB - Heart transplantation is the most effective therapy for end-stage heart failure in patients with diabetes mellitus (DM). However, diabetes-related complications (DRCs) are a relative contraindication for heart transplantation. Nevertheless, the increasing prevalence of both DM and congestive heart failure makes it necessary to perform heart transplantation even in those patients with advanced DM. We performed a retrospective analysis on long-term survival in 47 patients with insulin-treated DM and DRCs (group 1). Survival rate and causes of death were compared with data of a group of heart transplant recipients without DM (n = 1061, group 2). Mean follow-up time of all heart transplant recipients was 68.2 months (range: 0-204 months). Overall mortality during follow-up was 42.9%. Long term survival did not differ significantly between study groups, but tended to be shorter in group 1 than in group 2 (P = 0.07). In group 1, steroid-free immunosuppressive therapy was associated with a higher percentage of long-term survivors compared with no steroid-free immunosuppression. Our data demonstrate that long-term survival is acceptable in heart transplant recipients with preoperatively diagnosed DM and DRCs. Consequently, advanced DM should no longer be a relative contraindication for heart transplantation. PMID- 17403109 TI - The uremic solute indoxyl sulfate induces oxidative stress in endothelial cells. AB - BACKGROUND: Endothelial dysfunction and oxidative stress are matters of concern in patients with chronic renal failure (CRF). Uremic solutes retained in these patients could be involved in these processes. Notably, the protein-bound uremic solute indoxyl sulfate induces endothelial dysfunction in vitro, and has shown pro-oxidant effects. OBJECTIVE: To demonstrate that indoxyl sulfate is a potential mediator of oxidative stress in endothelial cells in vitro. METHODS: Indoxyl sulfate-induced oxidative stress in human umbilical vein endothelial cells (HUVEC) was studied by measuring reactive oxygen specie (ROS) production by cytofluorimetry, by analyzing the involvement of the pro-oxidative enzymes NAD(P)H oxidase, xanthine oxidase, and NO synthase, and by measuring the levels of the non-enzymatic antioxidant glutathione. RESULTS: We showed that indoxyl sulfate induced a significant production of ROS in HUVEC, with or without human serum albumin. We then investigated the role of pro-oxidative enzymes and measured the levels of the antioxidant glutathione. The NAD(P)H oxidase inhibitors, DPI, and apocynin, inhibited ROS production, whereas inhibitors of xanthine oxidase, NO synthase, and mitochondrial ROS had no effect. Interestingly, indoxyl sulfate strongly decreased the levels of glutathione, one of the most active antioxidant systems of the cell. In addition, the ROS production mediated by indoxyl sulfate was inhibited by the antioxidants vitamin C, vitamin E, and NAC. CONCLUSION: The uremic solute indoxyl sulfate enhances ROS production, increases NAD(P)H oxidase activity, and decreases glutathione levels in endothelial cells. Thus, indoxyl sulfate induces oxidative stress by modifying the balance between pro- and antioxidant mechanisms in endothelial cells. PMID- 17403110 TI - The intrinsic enzymatic activity of procarboxypeptidase U (TAFI) does not significantly influence the fibrinolysis rate: a rebuttal. PMID- 17403111 TI - A mechanism to safeguard platelet adhesion under high-shear flow: von Willebrand factor-glycoprotein Ib and integrin alpha2beta1-collagen interactions make complementary, collagen-type-specific contributions to adhesion: a rebuttal. PMID- 17403112 TI - Left ventricular systolic dysfunction is an independent predictor of homocysteine in angiographically documented patients with or without coronary artery lesions. AB - BACKGROUND: Elevated plasma homocysteine is a risk factor for coronary artery disease (CAD) and thromboembolic disorders that seems also to be associated with chronic heart failure. OBJECTIVE: To evaluate the association between homocysteine and left ventricular dysfunction and to assess whether it is independent of CAD. PATIENTS AND METHODS: A prospective study evaluated this relationship in 709 patients referred for diagnostic coronary angiography, including 515 CAD and 194 patients without evidence of coronary artery lesions. RESULTS: The homocysteine level was significantly higher in the 187 patients with a left ventricular ejection fraction (LVEF) dysfunction < 40% (P < 0.0001) than in those without ventricular dysfunction. LVEF, NYHA functional class II or III and CAD, stable angina and hypertension were clinical characteristics that influenced total homocysteine level in univariate analysis. Homocysteine was significantly associated with LVEF and N-terminal pro-B-type natriuretic peptide (NT-pro-BNP) in univariate regression (r = -0.267, 95% CI -0.33 to -0.19, P < 0.0001, and r = 0.381, 95% CI 0.28-0.47, P < 0.0001, respectively) and in multiple regression (P = 0.0022 and P = 0.0001, respectively). Other determinants were creatinine and vitamin B(12), but not folate. LVEF was a predictor of homocysteine > 15 micromol L(-1) in the whole population (P for trend < or = 0.0001) and in patients without documented CAD (P for trend = 0.0058). CONCLUSION: Our results showed an association of homocysteine with left ventricular systolic dysfunction and NT-pro-BNP that existed independently of documented CAD. Whether this association reflects a causative factor or a consequence of CHF and influences the prognosis of the disease remains an open question. PMID- 17403113 TI - Role of elevated alpha2-macroglobulin revisited: results of a case-control study in children with symptomatic thromboembolism. AB - OBJECTIVE: alpha(2)-Macroglobulin (alpha2MG) is a broad-spectrum protease inhibitor that is known to neutralize alpha-thrombin, plasmin, and activated protein C, which suggests that it has anticoagulant as well as procoagulant properties. The present study was conducted to evaluate the role of alpha2MG in children with venous thromboembolism [VTE: paradoxical embolism causing ischemic stroke (IS) or deep-vein thrombosis (DVT)]. METHODS: alpha2MG levels measured after acute VTE onset in white patients were compared with data obtained from age and gender-matched healthy controls. In addition, to compare the rate of elevated alpha2MG and prothrombotic risk factors [factor V G1691A, prothrombin G20210A, raised lipoprotein (a)] between patients and controls and to evaluate the interaction between elevated alpha2MG levels and other thrombophilias, odds ratios (ORs) together with 95% confidence intervals (CIs) were estimated using a logistic regression model. The model was adjusted for age and fibrinogen. RESULTS: alpha2MG levels were significantly higher in patients than in controls (320/139-524 vs. 302/109-406; P = 0.005). In the group of patients (IS n = 103; DVT n = 92), the risk of symptomatic thromboembolism was significantly increased with elevated alpha2MG levels, with a gradual increase per mg dL(-1). In addition, when elevated alpha2MG levels > 90th percentile were compared with values below the cut-off, including established prothrombotic risk factors in the multivariate analysis, patients had a significantly increased OR/95% CI for fibrinogen-adjusted alpha2MG levels (IS, 5.9/1.9-18.3; DVT, 7.2/2.1-24.4). CONCLUSIONS: The procoagulant properties of elevated alpha2MG levels independently increase the odds of stroke and DVT in white children. PMID- 17403114 TI - Glucose tolerance status and cognitive impairment in early middle-aged women. AB - AIMS: To examine the effect of glucose tolerance status on cognitive performance in early middle-aged women. METHODS: We conducted a matched, case-control study from a population-based cohort in Kinmen, Taiwan. Women aged 40-54 years were recruited. Participants were classified into one of three groups: normal, impaired glucose tolerance (IGT) or diabetes mellitus. Cognitive measures used were the Rey Auditory-Verbal Learning Test, visual memory, verbal fluency, Trail Making Test, and digit spans. RESULTS: Seven hundred and twenty eligible women underwent both the cognitive and oral glucose tolerance tests. The 72 women (10%) who had diabetes mellitus were compared with 68 women with IGT and 144 with normal glucose tolerance. All groups were matched for age and education level. Mean scores on cognitive testing did not differ between the three groups, except for the forward digit spans test. After adjustment for menopausal status, psychological stress scores and other cardiovascular variables, glucose tolerance status was no longer a significant predictor for forward digit span scores. CONCLUSIONS: Glucose tolerance status was not associated with cognitive performance in middle-aged women. PMID- 17403115 TI - The relationship of heart rate and heart rate variability to non-diabetic fasting glucose levels and the metabolic syndrome: the Cardiovascular Health Study. AB - BACKGROUND: Increased heart rate (HR) and diminished heart rate variability (HRV) are signs of early cardiovascular autonomic neuropathy. We tested the hypotheses that increased HR and diminished HRV are present in people: (i) with increased fasting glucose (FG) levels not in the range of diabetes mellitus (DM), and (ii) in people with the metabolic syndrome (MetS) independent of elevated FG levels. METHODS: HR and HRV were determined in 1267 adults (mean age 72 years) who had Holter monitoring and FG measures: 536 had normal FG levels (NORM, FG 4.5-5.5 mmol/l), 363 had mildly impaired FG (IFG-1, FG 5.6-6.0 mmol/l), 182 had significantly impaired FG (IFG-2, FG 6.1-6.9 mmol/l) and 178 had DM (FG > 6.9 mmol/l or use of glucose-lowering agents/insulin). HR and HRV in NORM/IFG-1 was further compared by the number of components of the MetS and compared by the presence or absence of MetS in IFG-2/DM. RESULTS: HRV indices were more impaired in IFG-2 and DM than in NORM or IFG-1. There were few differences in HRV indices between NORM and IFG-1 or between IFG-2 and DM. In NORM/IFG-1 participants, having > or = 2 components of the MetS was associated with a greater decrease in HRV compared with having no or one components. In IFG-2/DM participants, MetS was associated with decreased HRV compared with no MetS. CONCLUSIONS: Increased HR and diminished HRV occur in the non-diabetic FG range. Diminished HRV is associated with the MetS, independent of FG levels. Both these results suggest that factors associated with increasing non-diabetic FG levels and the MetS play a role in the onset of cardiac autonomic impairment. PMID- 17403116 TI - A national survey of in-patient diabetes services in the United Kingdom. AB - AIM: To examine in-patient diabetes services in all UK acute hospitals. METHODS: We asked the diabetes specialist team in all UK acute hospitals to complete a structured questionnaire on in-patient diabetes management guidelines, in-patient referral patterns, diabetes in-patient specialist nurse (DISN) services and diabetes bed occupancy in their hospital. RESULTS: Of the 262 UK acute hospitals, 239 (91.2%) provided data (2005-2006). UK teams reported high levels of clinical risk associated with in-patient diabetes care. One-third did not have diabetes management guidelines for day surgery, endoscopy, barium studies or immediate management of the diabetic foot. Patients admitted with diabetic ketoacidosis were not immediately referred to the specialist team in one-third of hospitals. About half had no routine access to podiatry or dietetic care for in-patients with diabetes. The majority of UK hospitals either never adopted Diabetes Mellitus, Insulin Glucose Infusion in Acute Myocardial Infarction (DIGAMI)-1 protocols or had recently changed practice, and half do not endorse the use of in patient subcutaneous insulin 'sliding-scales'. One in five UK hospitals survey in patient diabetes treatment satisfaction. DISN numbers have increased rapidly-126 hospitals (51.4%) had a DISN, most (69.1%) appointed since 2002. Most (80.2%) hospitals without a DISN used the out-patient specialist nurse team to provide in patient care. CONCLUSIONS: This survey has identified substantial gaps in in patient diabetes care in the UK. The rapid increase in DISN numbers indicates increasing attention to in-patient diabetes care in UK hospitals. PMID- 17403117 TI - No evidence for increased self-reported cognitive failure in Type 1 and Type 2 diabetes: a cross-sectional study. AB - AIMS: Mild cognitive deficits have been determined in both types of diabetes using neurocognitive tests. Little is known about the degree to which patients complain about their cognitive functioning. This study set out to investigate the magnitude and correlates of self-reported cognitive failure in adult out-patients with Type 1 and Type 2 diabetes. METHODS: Subjective cognitive functioning was measured in 187 diabetic patients using the Cognitive Failures Questionnaire (CFQ). Demographic and clinical characteristics were retrieved from the medical records. The Patient Health Questionnaire 9 items (PHQ-9) was self-administered along with the CFQ to correct for the confounding effect of depression. RESULTS: Analyses were based on 55 patients with Type 1 diabetes and 100 patients with Type 2 diabetes. No difference in mean CFQ score was observed between Type 1 and Type 2 diabetic patients or between Type 1 diabetic patients and healthy control subjects. Female patients with Type 2 diabetes reported significantly fewer cognitive complaints compared with female healthy control subjects. None of the demographic variables and diabetes-related complications was associated with subjective cognitive complaints. A strong positive association was found between depression symptomatology and frequency of self-reported cognitive failure. CONCLUSIONS: Our study could not confirm elevated subjective cognitive complaints in a group of Type 1 and Type 2 diabetes patients, as might be expected given the observed elevated rates of mild cognitive dysfunction in patients with diabetes. Self-reported cognitive failure appears largely determined by depressive symptomatology. Therefore, affective status should be included in any cognitive assessment procedure. PMID- 17403118 TI - Breastfeeding habits in families with Type 1 diabetes. AB - AIMS: Breastfeeding is acknowledged to be beneficial for child development. Women with diabetes may be more likely not to breastfeed their children because of neonatal morbidity and instability in diabetes control. The aim of this study was to assess the effect of maternal Type 1 diabetes on breastfeeding habits. METHODS: Full breastfeeding and any breastfeeding were reported in the first year of life in 1560 children born in Germany between 1989 and 2004. Of those, 997 children had a mother with Type 1 diabetes, and the remaining 563 children had a father or sibling with Type 1 diabetes. RESULTS: Fewer children of mothers with Type 1 diabetes were breastfed than children of non-diabetic mothers (77 vs. 86%; P < 0.0001) and, amongst breastfed children, there was a shorter duration of full breastfeeding (12 vs. 17 weeks; P < 0.0001) and any breastfeeding (20 vs. 26 weeks, P < 0.0001) in children of mothers with Type 1 diabetes compared with children of non-diabetic mothers. Other factors associated with reduced frequency and duration of breastfeeding were pre-term delivery (P < 0.0001), young maternal age (P < 0.0001), and firstborn children (P < 0.0001). After stratification for each of these factors, breastfeeding remained significantly less frequent and of less duration in children of mothers with Type 1 diabetes as compared with children of non-diabetic mothers. CONCLUSIONS: Mothers with Type 1 diabetes breastfeed their children less than international recommendations. Counselling to increase frequency and duration of breastfeeding may be warranted in this population. PMID- 17403119 TI - Perinatal factors associated with subsequent diabetes mellitus in the child: record linkage study. AB - AIMS: To report on associations between perinatal factors and the subsequent development of diabetes mellitus under the age of 30 years in the offspring. METHODS: Analysis of linked hospital statistical records, comparing perinatal factors relating to the birth of 518 people admitted to hospital for diabetes with the same factors in 292 845 others, in a defined population in southern England from 1963 to 1999. RESULTS: Diabetes mellitus was much more common in children of mothers with diabetes than in others (odds ratio 6.42; 95% confidence interval 4.18-9.86). There was no significant association with birthweight or gestational age separately. Diabetes was more common in those in the highest quintile of 'birthweight for gestational age' compared with the lowest four quintiles combined (odds ratio 1.33; 95% confidence interval 1.08-1.64), but there was no consistent gradient of increasing frequency of diabetes across the lowest four quintiles. There were no significant associations between diabetes and mothers' age, parity, social class, or smoking during pregnancy, or between babies' mode of delivery or any other perinatal factors investigated. All results were similar when the analysis was confined to diabetes in people aged < 15 years. CONCLUSIONS: We found a strong association between diabetes in the child mainly, if not entirely Type 1 diabetes-and maternal diabetes. Diabetes was slightly more common in the heaviest quintile of birthweight for gestational age than in other quintiles. There were no significant associations between diabetes and the other perinatal factors studied. PMID- 17403120 TI - Interactive effect of retinopathy and macroalbuminuria on all-cause mortality, cardiovascular and renal end points in Chinese patients with Type 2 diabetes mellitus. AB - AIMS: To examine the effect of albuminuria and retinopathy on the risk of cardiovascular and renal events, and all-cause mortality in patients with Type 2 diabetes. METHODS: A post-hoc analysis of 4416 Chinese patients without macrovascular complications at baseline (age 57.6 +/- 13.3 years). Glomerular filtration rate (eGFR) was estimated by the abbreviated Modification of Diet in Renal Disease Study Group Formula, further adjusted for Chinese ethnicity. Clinical end points were all-cause mortality, cardiovascular events (heart failure or angina, myocardial infarction, lower limb amputation, re vascularization procedures and stroke) and renal end points (reduction in eGFR by more than 50% or eGFR < 15 ml/min/1.73 m2 or death as a result of renal causes or need for dialysis). RESULTS: Compared with individuals without complications, subjects with retinopathy and macroalbuminuria had higher rates of cardiovascular events (14.1 vs. 2.4%), renal events (40.0 vs. 0.8%) and death (9.3 vs. 1.7%, P < 0.001). For composite event of death, cardiovascular and renal events, the presence of retinopathy, microalbuminuria alone, macroalbuminuria alone, retinopathy with microalbuminuria or retinopathy with macroalbuminuria increased the risk [hazard ratio (95% CI)] by 1.61 (1.05 to 2.47; P = 0.04), 1.93 (1.38 to 2.69; P < 0.001), 4.34 (3.02 to 6.22; P < 0.001), 2.59 [1.76 to 3.81; P < 0.001) and 6.83 (4.89 to 9.55; P < 0.001) fold, respectively. The relative excess risk as a result of interaction between retinopathy and macroalbuminuria was 15.31, implying biological interaction in the development of renal events. CONCLUSIONS: In Chinese patients with Type 2 diabetes, retinopathy interacts with macroalbuminuria to increase the risk of composite cardio-renal events. PMID- 17403121 TI - A double-blind randomized study comparing the effects of continuing or not continuing rosiglitazone + metformin therapy when starting insulin therapy in people with Type 2 diabetes. AB - AIMS: To compare the efficacy and safety of either continuing or discontinuing rosiglitazone + metformin fixed-dose combination when starting insulin therapy in people with Type 2 diabetes inadequately controlled on oral therapy. METHODS: In this 24-week double-blind study, 324 individuals with Type 2 diabetes inadequately controlled on maximum dose rosiglitazone + metformin therapy were randomly assigned to twice-daily premix insulin therapy (target pre-breakfast and pre-evening meal glucose < or = 6.5 mmol/l) in addition to either rosiglitazone + metformin (8/2000 mg) or placebo. RESULTS: Insulin dose at week 24 was significantly lower with rosiglitazone + metformin (33.5 +/- 1.5 U/day, mean +/- se) compared with placebo [59.0 +/- 3.0 U/day; model-adjusted difference -26.6 (95% CI -37.7, -15,5) U/day, P < 0.001]. Despite this, there was greater improvement in glycaemic control [HbA(1c) rosiglitazone + metformin vs. placebo 6.8 +/- 0.1 vs. 7.5 +/- 0.1%; difference -0.7 (-0.8, -0.5)%, P < 0.001] and more individuals achieved glycaemic targets (HbA(1c) < 7.0% 70 vs. 34%, P < 0.001). The proportion of individuals reporting at least one hypoglycaemic event during the last 12 weeks of treatment was similar in the two groups (rosiglitazone + metformin vs. placebo 25 vs. 27%). People receiving rosiglitazone + metformin in addition to insulin reported greater treatment satisfaction than those receiving insulin alone. Both treatment regimens were well tolerated but more participants had oedema [12 (7%) vs. 4 (3%)] and there was more weight gain [3.7 vs. 2.6 kg; difference 1.1 (0.2, 2.1) kg, P = 0.02] with rosiglitazone + metformin. CONCLUSIONS: Addition of insulin to rosiglitazone + metformin enabled more people to reach glycaemic targets with less insulin, and was generally well tolerated. PMID- 17403122 TI - Prevalence of depression and diabetes: a population-based study from rural Bangladesh. AB - AIMS: To determine the extent of depressive symptoms in a rural community of Bangladesh and its association with newly recognized diabetes. METHODS: Depressive symptoms were assessed in 184 newly diagnosed diabetic subjects and 768 randomly selected individuals without diabetes. The Montogomery and Aasberg Depression Rating Scale (MADRS) was used to assess depressive symptoms. A structured interview was performed to obtain socio-demographic and economic information and anthropometric measures were collected. Fasting plasma glucose was measured by the HemoCue glucose analyser. RESULTS: Twenty-nine percent of male and 30.5% of female participants with diabetes and 6.0% of male and 14.6% of female subjects without diabetes had depressive symptoms rating > or = 20 on the MADRS. An association between depressive symptoms and diabetes was found (P < 0.01). After controlling for potential confounding factors including age, gender, fasting plasma glucose > 7.0 mmol/l and waist-hip ratio, the association of depression with diabetes remained significant. CONCLUSIONS: An unexpectedly high level of unrecognized depressive symptoms was found in the general rural population of Bangladesh. These are among the first data to suggest that depressive symptoms in this culture are common, especially in women. Depression is particularly common in those with diabetes. Psychiatric intervention may be necessary in addition to lifestyle changes to prevent the exponential increase in the occurrence of Type 2 diabetes. In addition, a common approach including psychiatric treatment in diabetes care may be necessary to achieve improved glycaemic control in this population. PMID- 17403123 TI - Foot abnormalities in Canadian Aboriginal adolescents with Type 2 diabetes. AB - AIMS: To determine the profile of foot abnormalities in Canadian Aboriginal adolescents with Type 2 diabetes and the risk factors associated with these abnormalities. METHODS: Aboriginal adolescents with Type 2 diabetes underwent an interview, medical record review and foot examination in a tertiary care, paediatric hospital diabetes clinic and two geographically remote outreach clinics. The notes of 110 subjects were reviewed [mean age 15 +/- 3 years; mean duration of diabetes, 30 +/- 20 months; 71 (66%) female and 39 (34%) male] and 77 (70%) of the subjects were examined. RESULTS: Foot abnormalities were identified by either interview or notes review, and included poor toenail condition in 85 (77%), paronychia in 29 (26%), ingrowing toenails in 16 (15%) and neuropathic symptoms in 13 (12%) subjects. Foot abnormalities were identified by examination in many subjects, including poor toenail condition in 38 (49%), calluses in 34 (44%) and paronychia in 13 (17%) subjects. Eighteen (24%) of 75 subjects did not have running water in the home. Factors that significantly increased the presence of foot abnormalities included: foot care provided by a person other than self; absence of running water in the home; decreased frequency of bathing; and decreased frequency of nail clipping. A greater percentage of subjects living on a reservation or rural community had specialized consultations for retinal examination, footwear, or both than of those living in an urban or unknown residence. CONCLUSIONS: A high prevalence of foot abnormalities was noted in Aboriginal adolescents with Type 2 diabetes. These findings highlight the associated comorbidities in this population, emphasizing the need for early detection and intervention. PMID- 17403124 TI - Lack of association between polymorphisms in the gene encoding protein tyrosine phosphatase 1B (PTPN1) and risk of Type 2 diabetes. AB - AIMS: Recently, an association of Type 2 diabetes (T2DM) with polymorphisms in PTPN1 located on chromosome 20q was reported. We attempted to replicate this finding in an ethnically homogeneous Polish population. METHODS: The study groups comprised 474 cases with T2DM and 411 control subjects with normal fasting glucose. All individuals were genotyped for the five previously reported PTPN1 polymorphisms using a fluorescence polarization method. HAPLO.STAT software was used to infer and compare haplotype distributions. RESULTS: The distributions of alleles and genotypes for the five genotyped PTPN1 polymorphisms did not differ between the T2DM cases and control subjects (lowest P = 0.6). Similarly, the frequency of the common haplotype reported to be associated with T2DM did not differ in cases and control subjects. We also failed to find such an association in Whites by performing a meta-analysis of all the available data on the association of those five SNPs with T2DM. CONCLUSION: This case-control study in a Polish population did not confirm the reported association between polymorphisms in PTPN1 and T2DM. PMID- 17403125 TI - Development of an educational 'toolkit' for health professionals and their patients with prediabetes: the WAKEUP study (Ways of Addressing Knowledge Education and Understanding in Pre-diabetes). AB - AIMS: To identify key messages about pre-diabetes and to design, develop and pilot an educational toolkit to address the information needs of patients and health professionals. METHODS: Mixed qualitative methodology within an action research framework. Focus group interviews with patients and health professionals and discussion with an expert reference group aimed to identify the important messages and produce a draft toolkit. Two action research cycles were then conducted in two general practices, during which the draft toolkit was used and video-taped consultations and follow-up patient interviews provided further data. Framework analysis techniques were used to examine the data and to elicit action points for improving the toolkit. RESULTS: The key messages about pre-diabetes concerned the seriousness of the condition, the preventability of progression to diabetes, and the need for lifestyle change. As well as feedback on the acceptability and use of the toolkit, four main themes were identified in the data: knowledge and education needs (of both patients and health professionals); communicating knowledge and motivating change; redesign of practice systems to support pre-diabetes management and the role of the health professional. The toolkit we developed was found to be an acceptable and useful resource for both patients and health practitioners. CONCLUSIONS: Three key messages about pre diabetes were identified. A toolkit of information materials for patients with pre-diabetes and the health professionals and ideas for improving practice systems for managing pre-diabetes were developed and successfully piloted. Further work is needed to establish the best mode of delivery of the WAKEUP toolkit. PMID- 17403126 TI - A randomized controlled trial examining combinations of repaglinide, metformin and NPH insulin. AB - AIMS: To compare combination use of repaglinide, metformin and bedtime Neutral Protamine Hagedorn (NPH) insulin with conventional approaches of insulin initiation in patients with Type 2 diabetes (T2DM). METHODS: Eighty-two patients with T2DM with suboptimal glycaemic control on oral glucose-lowering agents were randomized to one of three treatment regimens for 4 months. Group 1 received metformin and twice daily biphasic 30/70 human insulin mixture (n = 27), group 2 metformin and bedtime NPH insulin (n = 26) and group 3 metformin, bedtime NPH insulin and mealtime repaglinide (n = 25). RESULTS: Seventy-five patients completed the study. Baseline and end-point mean HbA1c levels fell from 9.0 +/- 1.1 to 7.9 +/- 1.1% in group 1, 10.0 +/- 2.2 to 9.2 +/- 1.4% group 2 and 10.0 +/- 1.7 to 8.1 +/- 1.5% in group 3, respectively. All groups showed improvements in HbA1c. There was no significant difference between groups in the proportions of patients experiencing hypoglycaemia (29.6, 25.0 and 16.7%, respectively; P = 0.55) or in mean weight gain (2.9, 0.7 and 2.2 kg, respectively; P = 0.06). By 4 months, insulin doses were 0.63 +/- 0.32 IU/kg in group 1, 0.58 +/- 0.21 IU/kg in group 2 and 0.37 +/- 0.22 IU/kg in group 3 (group 3 vs. groups 1 and 2: P < 0.002). CONCLUSIONS: The approach using repaglinide, metformin and NPH insulin improved glycaemic control with a similar safety profile to conventional insulin initiation in T2DM and produced final glycaemic control similar to metformin and a twice daily biphasic insulin mixture. PMID- 17403127 TI - A new multiplex PCR for easy screening of methicillin-resistant Staphylococcus aureus SCCmec types I-V. AB - A multiplex PCR with four primer-pairs was designed to identify the five main known SCCmec types. A clear and easily discriminated band pattern was obtained for all five types. The SCCmec type was identified for 98% of 312 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). SCCmec type IV was by far the most common SCCmec type among both hospital- and community acquired MRSA isolates in Denmark. PMID- 17403128 TI - Detection of Fusobacterium necrophorum subsp. funduliforme in tonsillitis in young adults by real-time PCR. AB - Throat swabs from 61 patients, aged 18-32 years, with non-streptococcal tonsillitis (NST) and 92 healthy controls were examined for the presence of Fusobacterium necrophorum DNA using a novel TaqMan-based real-time quantitative PCR assay for F. necrophorum subspecies. The assay was based on the gyrB subunit gene, and detected F. necrophorum DNA in 48% of patients with NST and in 21% of controls (p <0.001). F. necrophorum subsp. funduliforme was the only subspecies found in both patients and controls. The load of F. necrophorum DNA on swabs from patients with NST was significantly higher than that on swabs from controls (p <0.001). Furthermore, patients with recurrent NST had a significantly higher load of F. necrophorum DNA compared to patients with acute NST (p 0.04). In addition, 26 patients with tonsillitis and group C streptococci (GCS) had a significantly higher load of F. necrophorum DNA compared to the NST group (p <0.001). It was concluded that F. necrophorum subsp. funduliforme is present in small numbers as part of the normal human throat flora, and that F. necrophorum in large quantities may cause tonsillitis, especially recurrent tonsillitis. In addition, the study suggests that the concomitant presence of GCS may aggravate F. necrophorum tonsillitis. PMID- 17403129 TI - Clonal spread of SCCmec type IV methicillin-resistant Staphylococcus aureus between community and hospital. AB - The staphylococcal chromosome cassette (SCC)mec types of 382 hospital-acquired methicillin-resistant Staphylococcus aureus (HA-MRSA) isolates in Taiwan were analysed over a 7-year period (1999-2005). There was an abrupt increase in SCCmec type IV in HA-MRSA during 2005. The molecular epidemiology of a subset (n = 69) of HA-MRSA isolates with SCCmec types III, IV or V was characterised and compared with that of community-acquired MRSA (CA-MRSA) (n = 26, collected during 2005). Pulsed-field gel electrophoresis revealed three major pulsotypes (A, B and C) and 15 minor clones. Pulsotypes B and C, which contained isolates carrying SCCmec types IV and V, respectively, included both CA-MRSA and HA-MRSA isolates. Among 24 toxin genes analysed, five genes had significant differential distribution between CA-MRSA and SCCmec type III HA-MRSA. Furthermore, among SCCmec type IV isolates, the seb gene was detected more commonly in HA-MRSA. Analysis of representative members of the three major pulsotypes by multilocus sequence typing revealed two sequence types (STs), namely ST239 (SCCmecIII) and ST59 (SCCmecIV or SCCmecV). This suggests that ST59:SCCmecIV, which is usually community-acquired, has become an important nosocomial pathogen in the hospital studied. PMID- 17403130 TI - Activity of telithromycin and comparators against isolates of Legionella pneumophila collected from patients with community-acquired respiratory tract infections: PROTEKT Years 1-5. AB - The in-vitro activity of telithromycin and comparator antibacterial agents was determined against clinical isolates of Legionella pneumophila collected in the PROTEKT surveillance study. In total, 133 isolates were collected between 1999 and 2004 from 13 countries (Australia, Belgium, Czech Republic, France, Germany, Hungary, Ireland, Italy, Japan, Portugal, Spain, Sweden and the USA). MICs were determined by broth microdilution. Telithromycin maintained activity between Year 1 (MIC(90) 0.015 mg/L) and Year 5 (MIC(90) 0.03 mg/L), as did the comparator antibacterial agents. Telithromycin appears to be a candidate for coverage of legionellosis in the empirical treatment of community-acquired respiratory tract infection. PMID- 17403131 TI - Identification of a new variable number tandem repeat locus in Mycobacterium ulcerans for potential strain discrimination among African isolates. AB - Intra-species discrimination in the highly clonal pathogen Mycobacterium ulcerans was studied in a diverse collection of isolates by PCR amplification of a short sequence repeat locus containing heterogeneous arrays of tri-nucleotide repeats with an ACC consensus pattern. Post-amplification analysis indicated excellent typeability and identified five M. ulcerans alleles, including a unique Angolan type differentiated for the first time among a genetically conserved cluster of African isolates. These results are consistent with previously investigated independent markers, and provide an additional locus for variable number tandem repeat-based typing of M. ulcerans. PMID- 17403132 TI - Population-based laboratory surveillance for tribe Proteeae isolates in a large Canadian health region. AB - The tribe Proteeae comprises the genera Proteus, Morganella and Providencia. Few studies have specifically investigated the epidemiology of infections caused by the Proteeae, and none has been conducted in a large non-selected population. The present study was a population-based laboratory surveillance in the Calgary Health Region (population 1.2 million), Canada during 2000-2005 that aimed to define the incidence, demographical risk-factors for acquisition and antimicrobial susceptibilities of Proteeae isolates. In total, 5047 patients were identified from whom Proteeae isolates were obtained (an annual incidence of 75.9/100 000), with females and the elderly being at highest risk. Incidence rates were 64.8, 7.7 and 3.4/100,000/year for the genera Proteus, Morganella and Providencia, respectively. Overall, 85% of infections were community-onset, and the overall rate of bacteraemic disease was 2.0/100,000. Compared with other species, Proteus mirabilis occurred at a much higher frequency, especially among females, and was less likely to be isolated from hospital-onset infections or to be part of a polymicrobial infection. Among isolates from community-onset infections, Providencia spp. were less likely to be from outpatients and more likely to be from nursing home residents. There were low overall rates of resistance to ciprofloxacin (4%) and gentamicin (5%), with Prot. mirabilis generally being the most susceptible. Members of the Proteeae were isolated frequently in both the community and hospital settings, but were infrequent causes of invasive disease. The occurrence, demographical risk-factors and microbiology of Proteeae isolates varied according to the individual species. PMID- 17403133 TI - Human cytomegalovirus-associated oral and maxillo-facial disease. AB - Human cytomegalovirus is a ubiquitous pathogen with protean clinical manifestations. After initial infection, the virus remains in a persistent state in the host. Immunity plays a pivotal role in counteracting its virulence, albeit intermittent virus shedding occurs in immunocompetent individuals. Should deficiencies in immunity occur, e.g., as a consequence of AIDS or iatrogenic immunosuppression, then virus replication and subsequent pathogenic manifestations ensue. In the oral and maxillo-facial region, the virus causes a wide variety of diseases, mainly atypical chronic ulcerations and sialadenitis. These morbidities are rarely reported and sometimes cause significant problems for clinicians. PMID- 17403134 TI - Association of specific mutations in katG, rpoB, rpsL and rrs genes with spoligotypes of multidrug-resistant Mycobacterium tuberculosis isolates in Russia. AB - Most multidrug-resistant (MDR) Mycobacterium tuberculosis isolates in Russia belong to the Beijing or Latino-American and Mediterranean (LAM) spoligotype families. The objective of this study was to investigate possible associations between genotype and the frequencies of mutations that confer drug resistance in a population that has two large families of circulating strains. Spoligotyping, IS6110 restriction fragment length polymorphism typing, and sequencing of the katG and rpoB genes, were performed for 217 consecutive MDR M. tuberculosis isolates from patients. The rpsL and rrs genes were also sequenced for selected streptomycin-resistant isolates. Of the 217 MDR isolates, 99 (46%) belonged to the LAM family, 92 (42%) to the Beijing family, 21 (10%) to the Haarlem family and four (2%) to the T family. There was one unique spoligotype. Mutations in the katG gene were identified in 207 (95%) isolates, all of which had mutations in codon 315. Mutations in the rpoB gene were identified in 200 (92%) isolates; 75% of LAM isolates carried a mutation in codon 516, whereas 71% of Beijing isolates carried a mutation in codon 531. In the 33 isolates resistant to streptomycin 50 mg/L, the 43AGG rpsL mutation was found in 27% of Haarlem, 75% of Beijing and 0% of LAM isolates, and rrs mutations were found in 17% (516C-->T) of Beijing and 100% (513A-->C) of LAM isolates. Overall, there appeared to be a correlation between the genotype and specific mutations conferring resistance to rifampicin or streptomycin in the Beijing and LAM families. The biological implications of this correlation remain to be explored. PMID- 17403135 TI - Arachidonic acid metabolism in brain physiology and pathology: lessons from genetically altered mouse models. AB - The arachidonic acid (AA) cascade involves the release of AA from the membrane phospholipids by a phospholipase A(2), followed by its subsequent metabolism to bioactive prostanoids by cyclooxygenases coupled with terminal synthases. Altered brain AA metabolism has been implicated in neurological, neurodegenerative, and psychiatric disorders. The development of genetically altered mice lacking specific enzymes of the AA cascade has helped to elucidate the individual roles of these enzymes in brain physiology and pathology. The roles of AA and its metabolites in brain physiology, with a particular emphasis on the phospholipase A(2)/cyclooxygenases pathway, are summarized, and the specific phenotypes of genetically altered mice relevant to brain physiology and neurotoxic models are discussed. PMID- 17403136 TI - Evidence of calcium- and SNARE-dependent release of CuZn superoxide dismutase from rat pituitary GH3 cells and synaptosomes in response to depolarization. AB - The antioxidant enzyme CuZn superoxide dismutase (SOD1) is secreted by many cell lines. However, it is not clear whether SOD1 secretion is only constitutive or can be regulated in an activity-dependent fashion. Using rat pituitary GH(3) cells that express voltage-dependent calcium channels and are subjected to Ca(2+) oscillations, we found that treatment with high K(+)-induced SOD1 release that was significantly higher than the constitutive secretion. Evoked SOD1 release was correlated with depolarization-dependent calcium influx and was virtually abolished by removal of extracellular calcium with EGTA or by pre-incubation of GH(3) cells with Botulinum toxin A that cleaves the SNARE protein SNAP-25. Immunofluorescence experiments performed in GH(3) cells and rat brain synaptosomes showed that K(+)-depolarization induced a marked depletion of intracellular SOD1 immunoreactivity, an effect that was again abolished in the absence of extracellular calcium or after treatment with Botulinum toxin A. Subcellular fractionation analysis showed that SOD1 was present in large dense core vesicles. These data clearly show that, in addition to the constitutive SOD1 secretion, depolarization induces an additional rapid calcium-dependent SOD1 release in GH(3) cells and in rat brain synaptosomes. This likely occurs through exocytosis from SOD1-containing vesicles operated by the SNARE complex. PMID- 17403137 TI - Dexamethasone suppresses monocyte chemoattractant protein-1 production via mitogen activated protein kinase phosphatase-1 dependent inhibition of Jun N terminal kinase and p38 mitogen-activated protein kinase in activated rat microglia. AB - Microglial cells release monocyte chemoattractant protein-1 (MCP-1) which amplifies the inflammation process by promoting recruitment of macrophages and microglia to inflammatory sites in several neurological diseases. In the present study, dexamethasone (Dex), an anti-inflammatory and immunosuppressive drug has been shown to suppress the mRNA and protein expression of MCP-1 in activated microglia resulting in inhibition of microglial migration. This has been further confirmed by the chemotaxis assay which showed that Dex or MCP-1 neutralization with its antibody inhibits the microglial recruitment towards the conditioned medium of lipopolysaccharide (LPS)-treated microglial culture. This study also revealed that the down-regulation of the MCP-1 mRNA expression by Dex in activated microglial cells was mediated via mitogen-activated protein kinase (MAPK) pathways. It has been demonstrated that Dex inhibited the phosphorylation of Jun N-terminal kinase (JNK) and p38 MAP kinases as well as c-jun, the JNK substrate in microglia treated with LPS. The involvement of JNK and p38 MAPK pathways in induction of MCP-1 production in activated microglial cells was confirmed as there was an attenuation of MCP-1 protein release when microglial cells were treated with inhibitors of JNK and p38. In addition, Dex induced the expression of MAP kinase phosphatase-1 (MKP-1), the negative regulator of JNK and p38 MAP kinases in microglial cells exposed to LPS. Blockade of MKP-1 expression by triptolide enhanced the phosphorylation of JNK and p38 MAPK pathways and the mRNA expression of MCP-1 in activated microglial cells treated with Dex. In summary, Dex inhibits the MCP-1 production and subsequent microglial cells migration to the inflammatory site by regulating MKP-1 expression and the p38 and JNK MAPK pathways. This study reveals that the MKP-1 and MCP-1 as novel mediators of biological effects of Dex may help developing better therapeutic strategies for the treatment of patients with neuroinflammatory diseases. PMID- 17403138 TI - S100B protein is released from rat neonatal neurons, astrocytes, and microglia by in vitro trauma and anti-S100 increases trauma-induced delayed neuronal injury and negates the protective effect of exogenous S100B on neurons. AB - S100B protein is found in brain, has been used as a marker for brain injury and is neurotrophic. Using a well-characterized in vitro model of brain cell trauma, we have previously shown that strain injury causes S100B release from neonatal rat neuronal plus glial cultures and that exogenous S100B reduces delayed post traumatic neuronal damage even when given at 6 or 24 h post-trauma. The purpose of the current studies was to measure post-traumatic S100B release by specific brain cell types and to examine the effect of an antibody to S100 on post traumatic delayed (48 h) neuronal injury and the protective effect of exogenous S100B. Neonatal rat cortical cells grown on a deformable elastic membrane were subjected to a strain (stretch) injury produced by a 50 ms displacement of the membrane. S100B was measured with an ELISA kit. Trauma released S100B from pure cultures of astrocytes, microglia, and neurons. Anti-S100 reduced released S100B to below detectable levels, increased delayed neuronal injury in traumatized cells and negated the protective effect of exogenous S100B on injured cells. Heat denatured anti-S100 did not exacerbate injury. These studies provide further evidence for a protective role for S100B following neuronal trauma. PMID- 17403139 TI - Flumazenil selectively prevents the increase in alpha(4)-subunit gene expression and an associated change in GABA(A) receptor function induced by ethanol withdrawal. AB - The actions of ethanol on gamma-aminobutyric acid type A (GABA(A)) receptors are still highly controversial issues but it appears that some of its pharmacological effects may depend on receptor subunit composition. Prolonged ethanol exposure produces tolerance and dependence and its withdrawal alters GABA(A) receptor subunit gene expression and function. Whereas benzodiazepines are clinically effective in ameliorating ethanol withdrawal symptoms, work in our laboratory showed that benzodiazepines also prevent, in vitro, some of the ethanol withdrawal-induced molecular and functional changes of the GABA(A) receptors. In the present work, we investigated the effects, on such changes, of the benzodiazepine receptor antagonist flumazenil that can positively modulate alpha(4)-containing receptors. We here report that flumazenil prevented both the ethanol withdrawal-induced up-regulation of the alpha(4)-subunit and the increase in its own modulatory action. In contrast, flumazenil did not inhibit ethanol withdrawal-induced decrease in alpha(1)- and delta-subunit expression as well as the corresponding decrease in the modulatory action on GABA(A) receptor function of both the alpha(1)-selective ligand zaleplon and the delta-containing receptor preferentially acting steroid allopregnanolone. These observations are the first molecular and functional evidence that show a selective inhibition by flumazenil of the up-regulation of alpha(4)-subunit expression elicited by ethanol withdrawal. PMID- 17403140 TI - Gap-junction blocker carbenoxolone differentially enhances NMDA-induced cell death in hippocampal neurons and astrocytes in co-culture. AB - The beneficial or detrimental role of gap junction communication in the pathophysiology of brain injury is still controversial. We used co-cultures of hippocampal astrocytes and neurons, where we identified homocellular astrocyte astrocyte and heterocellular astrocyte-neuron coupling by fluorescence recovery after photobleaching, which was decreased by the gap junction blocker carbenoxolone (CBX). In these cultures, we determined the cell type-specific effects of CBX on the excitotoxic damage caused by N-methyl-D-aspartate (NMDA). We determined in both astrocytes and neurons the influence of CBX, alone or together with NMDA challenge, on cytotoxicity using propidium iodide labeling. CBX alone was not cytotoxic, but CBX treatment differentially accelerated the NMDA-induced cell death in both astrocytes and neurons. In addition, we measured mitochondrial potential using rhodamine 123, membrane potential using the oxonol dye bis(1,3-diethylthiobarbituric acid)trimethine oxonol, cytosolic Ca(2+) level using fura-2, and formation of reactive oxygen species (ROS) using dihydroethidium. CBX alone induced neither an intracellular Ca(2+) rise nor a membrane depolarization. However, CBX elicited a mitochondrial depolarization in both astrocytes and neurons and increased the ROS formation in neurons. In contrast, NMDA caused a membrane depolarization in neurons, coinciding with intracellular Ca(2+) rise, but neither mitochondrial depolarization nor ROS production seem to be involved in NMDA-mediated cytotoxicity. Pre-treatment with CBX accelerated the NMDA-induced membrane depolarization and prevented the repolarization of neurons after the NMDA challenge. We hypothesize that these effects are possibly mediated via blockage of gap junctions, and might be involved in the mechanism of CBX-induced acceleration of excitotoxic cell death, whereas the CBX-induced mitochondrial depolarization and ROS formation are not responsible for the increase in cytotoxicity. We conclude that both in astrocytes and neurons gap junctions provide protection against NMDA-induced cytotoxicity. PMID- 17403141 TI - Mitochondrial dysfunction early after traumatic brain injury in immature rats. AB - Mitochondria play central roles in acute brain injury; however, little is known about mitochondrial function following traumatic brain injury (TBI) to the immature brain. We hypothesized that TBI would cause mitochondrial dysfunction early (<4 h) after injury. Immature rats underwent controlled cortical impact (CCI) or sham injury to the left cortex, and mitochondria were isolated from both hemispheres at 1 and 4 h after TBI. Rates of phosphorylating (State 3) and resting (State 4) respiration were measured with and without bovine serum albumin. The respiratory control ratio was calculated (State 3/State 4). Rates of mitochondrial H(2)O(2) production, pyruvate dehydrogenase complex enzyme activity, and cytochrome c content were measured. Mitochondrial State 4 rates (ipsilateral/contralateral ratios) were higher after TBI at 1 h, which was reversed with bovine serum albumin. Four hours after TBI, pyruvate dehydrogenase complex activity and cytochrome c content (ipsilateral/contralateral ratios) were lower in TBI mitochondria. These data demonstrate abnormal mitochondrial function early (10 mg/L were excluded from the analysis. The remaining 65 patients (male/female, 41/24; age 49.1+/-11.4 years; dialysis duration 99.7+/-63.0 months) were studied. These patients had moderate malnutrition and the average MIS was 7.4 (range 3-17). The average weekly dose of administered rHuEPO was 69.1+/-63.1 U/kg. Malnutrition inflammation score had a positive correlation with the serum concentration of tumor necrosis factor-alpha, whereas it had a negative correlation with anthropometric measures, total iron-binding capacity, prealbumin, phosphorus, creatinine, and triglyceride. According to Pearson's correlation analysis, significant relationships of increased MIS with increased required rHuEPO dose and rHuEPO responsiveness index (EPO divided by hematocrit) were observed (p=0.008, r=-0.326; p=0.017, r=-0.306, respectively). Recombinant human erythropoietin dose requirement is correlated with MIS and adverse nutritional status in MHD patients without evidence of inflammation. Further research should focus on reversing the undergoing microinflammation for a better outcome in dialysis patients. PMID- 17403172 TI - Quality of life of end-stage renal disease patients and study on the implementation of nocturnal home hemodialysis in Greece. AB - The aim of this study was to measure the overall health-related quality of life of patients receiving hemodialysis in Greece and to assess willingness, motivation, and concerns about participating in a nocturnal home hemodialysis program. This review measured successfully the quality of life of 146 end-stage renal disease (ESRD) patients undergoing hemodialysis in 10 hemodialysis centers all over Greece and studied the factors that affect their choice to adopt a specific treatment. Physical and mental health status and health-related quality of life, measured by the Kidney Disease Quality of Life Short Form accompanied by the demographic data gathered, provide information that can help to improve ESRD patient care in Greece. A comparison with the Greek general population identifies lower physical functioning scores for the ESRD patients, while the mental functioning scores are comparable with those of the Greek general population. Moreover, it is important to notice the willingness expressed by half of the patients to participate in a potential program of nocturnal home hemodialysis. PMID- 17403173 TI - Uremic hyperhomocysteinemia: a randomized trial of folate treatment for the prevention of cardiovascular events. AB - Homocysteine is a risk factor for atherosclerosis in the general population, and serum homocysteine levels are almost universally elevated in chronic renal failure patients. When such patients are treated with dialysis, cardiovascular disease accounts for more than 50% of their mortality, which, in some proportion, may be pathophysiologically related to the elevated serum homocysteine levels. From April 2003 to March 2005, we conducted a 2-year, double-blind, randomized, placebo-controlled trial of 186 patients with end-stage kidney disease due to any cause, who were older than 18 years and stable on hemodialysis. Patients were assigned to receive either oral folic acid 10 mg 3 times a week immediately after every dialysis session under nurse supervision or an identical-appearing placebo for the entire study. On admission, plasma total homocysteine (tHcy) levels were above 13.9 micromol/L in 96.7% of patients (median 25.0 micromol/L, range 9.3 104.0 micromol/L). In the placebo group, tHcy levels remained elevated at 6, 12, and 24 months, while oral folate significantly decreased tHcy to a median value of 10.5 (2.8-20.3) micromol/L, (p<0.01). During the study, 38 patients (folic acid group 17 vs. placebo group 21; p=0.47) died from cardiovascular disease. Kaplan-Meier life table analysis dealing with the incidence of cardiovascular events, both fatal and nonfatal (myocardial infarction, arrhythmias, angina, heart failure, cerebrovascular accident), showed that 2 years of folic acid treatment and the lowering of the homocysteine blood levels had no effect on cardiovascular events (p=0.41; hazard ratio 1.24, 95% CI 0.74-2.10). However, the carotid artery intima-media wall thickness measured in a blinded fashion decreased from 1.94 +/- 0.59 mm to 1.67 +/- 0.38 mm (p<0.01) after 2 years of folate therapy. In this short-term study of uremic patients, 2 years of folic acid supplementation normalized the tHcy blood levels in 92.3% of patients but did not change the incidence of cardiovascular events compared with the control group. However, ultrasonography of the common carotid arteries performed at entry and 24 months later showed a significant decrease in intima-media thickness with folate supplementation. This suggests that early folate supplementation may benefit patients with chronic renal failure by preventing cardiovascular deterioration. PMID- 17403174 TI - Comparison between different dialysate calcium concentrations in nocturnal hemodialysis. AB - Benefits of dialysate with greater calcium (Ca) concentration are reported in nocturnal hemodialysis (NHD) to prevent Ca depletion and subsequent hyperparathyroidism. Studies with patients dialyzing against 1.25 mmol/L Ca baths demonstrate increases in alkaline phosphatase (ALP) and parathyroid hormone (PTH) and increasing dialysate Ca subsequently corrects this problem. However, whether 1.5 or 1.75 mmol/L dialysate Ca is most appropriate for NHD is yet to be determined, and differences in the effect on mineral metabolism of daily vs. alternate daily NHD have also not been well defined. We retrospectively analyzed mineral metabolism in 48 patients, from 2 institutions (30 at Monash and 18 at Geelong), undergoing home NHD (8 hr/night, 3.5-6 nights/week) for a minimum of 6 months. Thirty-seven patients were dialyzed against 1.5 mmol/L Ca bath and 11 patients against 1.75 mmol/L. We divided patients into 4 groups, based on dialysate Ca and also on the hours per week of dialysis, <40 (1.5 mmol/L, n=29 and 1.75 mmol/L, n=8) or > or =40 (n=4 and 7). We compared predialysis and postdialysis serum markers, time-averaged over a 6-month period, and the administration of calcitriol and Ca-based phosphate binders between 1.5 and 1.75 mmol/L Ca dialysate groups. Baseline characteristics between all groups were similar, with a slightly longer, but nonsignificant, duration of NHD in both 1.75 mmol/L dialysate groups compared with 1.5 mmol/L. The mean predialysis Ca, phosphate, and Ca x P were similar between the 1.5 and 1.75 mmol/L groups, regardless of NHD hr/week. Postdialysis Ca was significantly greater, with 1.75 vs. 1.5 mmol/L in those dialyzing <40 hr/week (2.64+/-0.19 vs. 2.50+/-0.12 mmol/L, p=0.046), but postdialysis Ca x P were similar (2.25+/-0.44 vs. 2.16+/ 0.29 mmol(2)/L(2), p=0.60). Parathyroid hormone was also lower with 1.75 vs. 1.5 mmol/L baths in the <40 hr/week groups (31.99+/-26.99 vs. 14.47+/-16.36 pmol/L, p=0.03), although this difference was not seen in those undertaking NHD > or =40 hr/week. Hemoglobin, ALP, and albumin were all similar between groups. There was also no difference in vitamin D requirement when using 1.75 mmol/L compared with the 1.5 mmol/L dialysate. Multivariate analysis to determine independent predictors of postdialysis serum Ca showed a statistically significant positive association with predialysis Ca, dialysate Ca, and total NHD hr/week. An elevated dialysate Ca concentration is required in NHD to prevent osteopenia but differences in serum markers of mineral metabolism between 1.5 and 1.75 mmol/L Ca dialysate in NHD in our study were few. This was similar for patients undertaking NHD <40 or > or =40 hr/week, although differences in the frequency of NHD may also be as important as dialysate Ca with regard to serum Ca levels. With concerns that prolonged higher Ca levels contribute to increased cardiovascular mortality, the optimal Ca dialysate bath is still unknown and further studies addressing bone metabolism with larger NHD numbers are required. PMID- 17403175 TI - Daily home hemodialysis at a health maintenance organization: three-year experience. AB - Daily home hemodialysis (DHD), 5 to 7 short-duration hemodialysis treatments per week, promotes self-care and has beneficial effects on a number of clinical outcomes including blood pressure and volume control, electrolyte balance, uremic symptoms and sequelae, and quality of life. We sought to demonstrate that DHD is feasible and confers clinical benefits that permit savings in overall healthcare costs despite expenditures on program infrastructure and supplies. We examined the following outcomes monthly for all patients: laboratory values, dialysis adequacy, hospital admission records, surgical and interventional radiology records, and prescription medication usage. Twelve patients completed training in our home hemodialysis unit between April 2003 and April 2006. The mean age at the time of training was 58 years and mean vintage was 62 months. The mean treatment time was 147 min, and the mean number of treatments performed was 5.3 per week. When 1 patient with morbid obesity was excluded due to intentional weight loss, the mean dry weight at initiation of training was 71.9+/-12.4 kg and increased to 74.3+/-12.4 kg by the end of the study (p=0.66). The mean albumin increased from a baseline of 3.9+/-0.3 to 4.3+/-1.1 gm/dL during DHD (p=0.0015). The mean serum phosphorus levels were 5.4+/-1.4 mg/dL. Phosphate binder usage increased from a mean baseline of 2.6+/-1.4 to 4.2+/-2.6 tablets per meal during DHD (p=0.08). The mean delivered single pool Kt/V was 0.87 per treatment. During the 234 months studied, there were 11 hospital admissions (0.56 admissions per patient per year), with a mean length of stay of 3.7 days. Our results demonstrate that DHD improves nutritional status and decreases hospital admissions for dialysis dependent patients. PMID- 17403176 TI - Volumetric hemodynamic parameters to guide fluid removal on hemodialysis in the intensive care unit. AB - Estimation of removable excess body fluid is difficult in critically ill patients with renal failure. Volumetric hemodynamic parameters are increasingly being used to guide fluid therapy in the intensive care unit, but their suitability to monitor fluid removal with hemodialysis in critically ill patients is not known. Changes in the extravascular lung water index (EVLWI) and intrathoracic blood volume index (ITBVI) measured with transpulmonary thermodilution immediately before and after hemodialysis were analyzed from 39 hemodialysis sessions of 9 patients consecutively treated in the medical intensive care unit of a German University Hospital. Additional hemodynamic, ventilation, and oxygenation-related parameters were recorded at the same time. Online relative blood volume (RBV) monitoring was performed in 29 sessions. Comparisons of pre and postdialysis values showed a significant reduction of the EVLWI with fluid removal (p=0.009), with only a slight nonsignificant decrease in the ITBVI. The cardiac index (CI) also decreased significantly (p=0.010), whereas blood pressure remained stable. Oxygenation improved significantly (p=0.005), and the hematocrit increased significantly with dialysis (p=0.039). There was no correlation between hematocrit changes and RBV measurements. Significant correlations existed between ITBVI and CI changes (p<0.001), but not to EVLWI reduction. The removal of excess body fluid on hemodialysis is reflected by the EVLWI reduction, whereas the preservation of cardiac preload is shown by ITBVI stability. Volumetric hemodynamic parameters could be useful to guide fluid removal with hemodialysis in the intensive care unit. PMID- 17403177 TI - Functional iron deficiency in hemodialysis patients with high ferritin. AB - Although functional iron deficiency (FID) may be present in hemodialysis (HD) patients with high serum ferritin levels (>800 ng/mL), current protocols often preclude the use of intravenous (IV) iron in these patients. However, it has not been demonstrated that iron supplementation during erythropoietin therapy is ineffective or unsafe in increasing hemoglobin (Hb) levels in patients with high serum ferritin. This report describes the hematologic efficacy and safety of ferric gluconate (FG) therapy in patients with serum ferritin >800 ng/mL. A retrospective analysis was performed on HD patients at a single California dialysis center from January 1 to December 31, 2003. Patients classified as having high ferritin levels (serum ferritin >800 ng/mL on at least 66% of routine monthly measurements and transferrin saturation [TSAT] <25% on at least 1 occasion) were stratified as follows: patients in Group I were suspected of having FID and received FG > or =250 mg IV over a 3-month period when Hb was <11 g/dL, and patients in Group II were thought not to have FID and received <250 mg FG over a 3-month period. Both groups received standard recombinant human erythropoietin therapy as per the unit's protocol. Of 496 patients, 95 exhibited high ferritin and of these, 39 patients had sufficient data for analysis. Group I patients (n=14) showed a significant increase in Hb levels compared with Group II (n=25). There was no increase in ferritin levels in response to iron administration. No significant differences in hospitalizations or infections were observed between groups. Hemodialysis patients with high ferritin levels may have FID, and IV iron therapy safely improves FID in some patients. A larger randomized trial examining the optimal management of iron administration in HD patients with high ferritin levels is warranted. PMID- 17403178 TI - A case management model to improve hemodialysis outpatient outcomes. AB - Optimal outpatient dialysis care is often difficult to achieve and a case management model to augment conventionally applied nursing and physician resources focusing on continuous quality improvement presents a possible solution to improving outcomes in this setting. We applied this model to patients followed by our physician group. Continuous quality improvement data generated from the dialysis unit database were used to analyze outcomes in patients enrolled in this model. Data from the cohort of patients followed in 2003 served as the reference source for comparative purposes. The nurse case manager assumed responsibility during the second quarter of 2004. Comparing outcomes data from 2005 with data from 2003, we were able to achieve a 3.12% improvement in the annualized mean percent crude mortality per 100 patient years (p<0.003). There was a 3.46-day trend to improvement in patient hospital days per year (p<0.06). The percentage of catheters used as primary access decreased by 9.59% (p<0.025), and the percentage of patients meeting an eKdrt/V goal > or =1.2 increased by 15.33% to 92.37% (p<0.001). These data appear to support the utility of a case manager model in our system. PMID- 17403179 TI - Chronic maintenance hemodialysis: making sense of the terminology. AB - During the early decades, the hemodialysis (HD) terminology for modality, technique and function altered little as the widely accepted regime of thrice weekly, 4-hourly dialysis varied little. In the last decade, however, a wide range of new options have emerged in all facets of HD therapy. This has led to a sudden expansion in terminology, some duplicating, some contradictory, some superfluous. The definitions used in 1 geographical region may mean something entirely different elsewhere, increasing cross-continental misunderstanding and misinterpretation and raising the often-asked question: "What exactly did the authors mean by that?" Although clearly the definitions used in this paper are also only the authors' opinion, we have sought to explore the use and sometimes confusing application of many commonly used terms, and we propose a number of possible deletions. Finally, we offer a descriptive data set that we believe should be used for all HD-related papers. Our conclusions will not always be welcomed--particularly by those who use terms we have rejected. Despite this, we believe it pertinent to fully review the dialysis terminology we use. Primarily, we hope to stimulate debate about which terms should be globally adopted and what those terms should mean when used. Although not all will agree with our conclusions, we hope this paper may provide a framework for a more streamlined, efficient, and globally acceptable nomenclature. PMID- 17403180 TI - Optimal timing of initiation of chronic hemodialysis? AB - Despite the availability of clinical guidelines for the timing of dialysis initiation in both the United States and Canada, patients continue to start dialysis at very low levels of predicted glomerular filtration rate (GFR). A cross-sectional study was performed to determine the demographic and clinical characteristics of patients who started hemodialysis, their level of GFR, and mortality at 1 and 2 years following the initiation of dialysis. Retrospective data were collected on all eligible patients who commenced chronic hemodialysis in 1 tertiary care center in Canada from March 2001 to February 2005. Only those patients who had been followed by a nephrologist in the chronic kidney disease clinic before dialysis initiation were included (n=271). Seventeen percent of patients started hemodialysis late (GFR<5 mL/min/1.73 m(2)). Compared with the group of patients who started dialysis earlier, the late start group were significantly younger (p=0.008), had more females (p=0.013), more employed (p=0.051), less cardiac (p<0.001), and peripheral vascular disease (p=0.031), and were taking medication for hypertension (p=0.041). Serum albumin was lower in the late start group (p=0.023). At year 1, there was no difference in mortality rate while at year 2, the earlier the dialysis, the greater the mortality rate (p=0.022). After adjustment for demographic variables and comorbidities, only antihypertensive use had an independent but weak association with the 2 year mortality. Adjustment for all these variables eliminated the significant association noted for the 2 year mortality in the early versus late dialysis start. The survival benefit for late versus early dialysis start appears to be multifactorial and relates to a preponderance of clinical and demographic factors favoring a lengthened survival occurring in the late dialysis group. Our survival benefit findings suggest the premorbid health condition is a more important determinant of 2 year survival than the timing of dialysis initiation. PMID- 17403181 TI - Current concepts in the pathogenesis of nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is an increasingly recognized cause of chronic liver disease, representing the leading cause of hepatology referral in some centres. However, its pathophysiology is not completely understood. Insulin resistance is one of the major mechanisms involved in disease prevalence and progression. Owing to the lack of an effective pharmacological therapy, recommendations on treatment are scarce and are based mainly on lifestyle changes, including diet and exercise. A review of the current literature on pathogenesis of NAFLD is presented in this article. PMID- 17403182 TI - Liver fibrosis: searching for cell model answers. AB - Hepatic stellate cells (HSC) are the principal fibrogenic cell type in the liver. Progress in understanding the cellular and molecular basis for the development and progression of liver fibrosis could be possible by the development of methods to isolate HSC from rodents and human liver. Growth of stellate cells on plastic led to a phenotypic response known as activation, which paralleled closely the response of these cells to injury in vivo. Actually, much of the current knowledge of stellate cell behaviour has been gained through primary culture studies, particularly from rats. Also, different laboratories that have established hepatic stellate cell lines from rats and humans have provided a stable and unlimited source of cells that express specific functions, making them suitable for culture-based studies of hepatic fibrosis. From these in vitro models grew a large body of information characterizing stellate cell activation, cytokine signalling, intracellular pathways regulating liver fibrogenesis, production of extracellular matrix proteins and development of antifibrotic drugs. PMID- 17403183 TI - Sphingolipid signalling and liver diseases. AB - Sphingolipids (SLs) comprise a class of lipids with important structural functions and increasing relevance in cellular signalling. In particular, ceramide has attracted considerable attention owing to its role as a second messenger modulating several cell functions such as proliferation, gene expression, differentiation, cell cycle arrest and cell death. Increasing evidence documents the role of SLs in stress and death ligand-induced hepatocellular death, which contributes to the progression of several liver diseases including steatohepatitis, ischaemia-reperfusion liver injury or hepatocarcinogenesis. Furthermore, recent data indicate that the accumulation of SLs in specific cell subcompartments, characteristic of many sphingolipidoses, contributes to the hepatic dysfunctions that accompany these inherited diseases. Hence, the regulation of the cell biology and metabolism of SLs may open up a novel therapeutic avenue in the treatment of liver diseases. PMID- 17403184 TI - Tacrolimus for the treatment of primary sclerosing cholangitis. AB - BACKGROUND: Results from a pilot investigation with tacrolimus for primary sclerosing cholangitis (PSC) demonstrated biochemical improvement without excessive drug toxicity. To date, no confirmatory study has been performed. AIMS: We sought to determine the safety and efficacy of tacrolimus in PSC. METHODS: An open-label, phase II study of tacrolimus 0.05 mg/kg twice daily for 1 year was performed. Target whole-blood concentrations ranged between 3 and 7 ng/ml. RESULTS: A total of 16 patients were enrolled. The median age was 50 years (range, 28-68), with 31% being women. The median serum alkaline phosphatase was 903 U/l, AST 88 U/l, total bilirubin 0.9 mg/dl, and albumin 3.8 g/dl. Based primarily on drug-related adverse events, only eight (50%) patients completed 1 year of therapy. After 1 year of therapy, however, significant improvements in median serum alkaline phosphatase (903 vs. 483, P=0.0001) and AST levels (88 vs. 78, P=0.002) were observed in these patients. The median tacrolimus level in patients completing 1 year of therapy was 4.0 ng/ml. Drug-related adverse events, however, were responsible for 31% of participants withdrawing from the study. CONCLUSIONS: Despite significant improvements in serum alkaline phosphatase, oral tacrolimus is poorly tolerated in patients with PSC. PMID- 17403185 TI - A novel therapeutic drug (copper nicotinic acid complex) for non-alcoholic fatty liver. AB - BACKGROUND: Fatty liver is the accumulation of fat in liver cells, which leads to disruption of the normal liver structure and function. METHODS: A non-alcoholic fatty liver rat model received copper (Cu) (I)-nicotinate complex [CuCl(HNA)2] for 4 weeks. RESULTS: Clinical signs and histopathological examinations showed obvious improvements in rats that received Cu complex who were continuously on an (HCFF) diet than those returned to standard diet with Cu complex. The improvement was matched in total lipids in sera and hepatic tissue, with disappearance of fat droplets from liver sections. Furthermore, the gain in body weight and the corresponding decrease in liver weight, decreased liver transaminases and alkaline phosphatase were prominent. The oxidative stress markers such as nitric oxide, lipid peroxides, glutathione and superoxide dismutase were obviously changed to healthy normal levels. CONCLUSION: The Cu complex may serve as a novel chemical restoring agent in fatty degenerated liver cells and for renewal of their structure and functions. However, clinical trials are required for more evaluation of the Cu complex in humans. PMID- 17403186 TI - Drug-induced liver injury at an Asian center: a prospective study. AB - BACKGROUND/AIMS: The aetiology of drug-induced liver injuries (DILI) in Asia is different from that in the West, as anecdotal studies have shown that traditional complementary and alternative medicines (CAM) accounted for a major proportion of offending drugs in DILI in Asia. We aimed to study DILI in Asia prospectively, and to test whether DILI caused by traditional CAM was related to adulterants. METHODS: A collaborative group consisting of a tertiary-hospital hepatology department, a pharmaceutical laboratory, and a pharmacovigilance unit was formed to study patients with DILI at a tertiary hospital over a 26-month period prospectively. Traditional medicines that were implicated were tested for the presence of adulterants. RESULTS: Thirty-one patients with DILI were enrolled: age 51+/-3 (18-79) years, 17 (55%) male. Twenty-three (74%) had hepatocellular, six (19%) had cholestatic, and two (7%) had a mixed pattern of injury. Chinese traditional CAM was the most common medication type implicated, accounting for 17 (55%) patients, followed by Malay CAM in five (16%). Thirty-one traditional medicines from 17 patients were available for chemical analysis. Adulterants were found in nine (29%) of them. CONCLUSIONS: DILI in Asia has a different aetiology as compared with the West, and could be related to presence of adulterants in traditional CAM. PMID- 17403187 TI - Artificial liver support with the molecular adsorbent recirculating system: activation of coagulation and bleeding complications. AB - BACKGROUND: Numerous, mostly uncontrolled, observations suggest that artificial liver support with the Molecular Adsorbent Recirculating System (MARS) improves pathophysiologic sequelae and outcome of acute and acute-on-chronic liver failure. MARS is felt to be safe, but extracorporeal circuits may activate coagulation. OBJECTIVE: To assess the frequency of and risk factors for activation of coagulation during MARS treatment. PATIENTS/METHODS: Retrospective analysis of coagulopathy/bleeding complications observed during 83 consecutive MARS sessions in 21 patients (11 men; median age 46 years; median three sessions per patient; median duration of session 8 h). RESULTS: Nine clinically relevant episodes of coagulopathy/bleeding were observed in eight patients, forced to premature cessation of MARS in seven and ended lethal in four. Four complications occurred during the first, five during later (third to seventh) MARS sessions and two bleeders tolerated further sessions without complications. Coagulation parameters worsened significantly also during MARS sessions not associated with bleeding (P< or =0.004). In univariate analysis, patient's age, vasopressor therapy, pretreatment INR, fibrin D-dimer and fibrinogen concentrations, but not severity of underlying disease (MELD, Child-Pugh, SAPS II scores), were significantly associated with coagulopathy (P<0.05). Only patient's age, fibrin D dimer level and INR were retained in a multivariate model correctly classifying 98% of sessions without, but only 33% with complications. CONCLUSION: Coagulation is frequently activated during MARS therapy, potentially leading to bleeding complications and mortality. PMID- 17403188 TI - Rotor-type hyperbilirubinaemia has no defect in the canalicular bilirubin export pump. AB - BACKGROUND: The cause of Rotor syndrome (RS), a rare-familial conjugated hyperbilirubinaemia with normal liver histology, is unclear. We hypothesized that RS can be an allelic variant of Dubin-Johnson syndrome, caused by mutation in ABCC2, and investigated ABCC2 (gene) and ABCC2 (protein) in two patients with RS. METHODS: A 57-year-old male presented with a 5-year history of predominantly conjugated hyperbilirubinaemia (170 micromol/l). Urinary porphyrin excretion was increased; cholescintigraphy revealed no chromoexcretion. A 68-year-old male presented with lifelong conjugated hyperbilirubinaemia (85 micromol/l). Bromosulfophthalein elimination was typical for RS. Both patients had histologically normal liver, without pigment. ABCC2 expression was investigated by confocal fluorescence microscopy. ABCC2 was sequenced from genomic DNA and cDNA, and exon deletions/duplications were sought by comparative genomic hybridization on a custom micro-array. RESULTS: In both patients, ABCC2 was expressed unremarkably at the apical membrane of hepatocytes and no sequence alterations were found in 32 exons, adjacent intronic regions and the promoter region of ABCC2. CONCLUSIONS: Rotor-type hyperbilirubinaemia is not an allelic variant of ABCC2 deficiency. PMID- 17403189 TI - Serum osteoprotegerin and its ligand in cirrhotic patients referred for orthotopic liver transplantation: relationship with metabolic bone disease. AB - AIMS: A prospective study was carried out in 22 cirrhotic patients referred for orthotopic liver transplantation, in order to analyze serum osteoprotegerin (OPG) and RANKL levels and their relationship with metabolic bone disease. METHODS: Serum levels of OPG and RANKL were measured in all patients as well as bone markers, serum parathyroid hormone and 25-hydroxyvitamin D levels. OPG and RANKL values were compared with those obtained in 29 healthy controls. Bone mineral density (BMD) of the lumbar spine and proximal femur was measured by dual X-ray absorptiometry and spinal X-rays were obtained to assess vertebral fractures. RESULTS: Serum OPG levels were higher in cirrhotic patients than in controls (6.4+/-2 vs 2.7+/-0.7 pmol/l; P=0.001) and RANKL serum levels were lower in cirrhotic patients (0.215+/-0.6 vs 1.012+/-1.2 pmol/l; P=0.002), with an increased OPG:RANKL ratio when compared with the control group (280.3+/-334.5 vs 113+/-137.6; P=0.04). Ten patients had osteoporosis (45%) and up to 45% skeletal fractures. No differences were found in OPG levels between patients with and without osteoporosis by densitometric criteria or fractures. Negative correlations were found between OPG levels and femoral neck (R-0.46; P=0.03) and total hip BMD (R-0.48; P=0.025). By contrast, OPG values were not related to markers of bone turnover. CONCLUSIONS: OPG values are elevated in cirrhotic patients before liver transplantation, particularly in those with low bone mass at the proximal femur. PMID- 17403190 TI - Model for end-stage liver disease score to serum sodium ratio index as a prognostic predictor and its correlation with portal pressure in patients with liver cirrhosis. AB - BACKGROUND: The models for end-stage liver disease (MELD) and serum sodium (SNa) are important prognostic markers in cirrhosis. A novel index, MELD to SNa ratio (MESO), was developed to amplify the opposing effect of MELD and SNa on outcome prediction. METHODS: A total of 213 cirrhotic patients undergoing hepatic venous pressure gradient (HVPG) measurement were retrospectively analyzed. RESULTS: The MESO index correlated with HVPG (r=0.258, P<0.001) and Child-Pugh score (rho=0.749, P<0.001). Using mortality as the end point, the area under receiver operating characteristic curve (AUC) was 0.860 for SNa, 0.795 for the MESO index and 0.789 for MELD (P values all >0.3) at 3 months. Among patients with Child Pugh class A or B, the MESO index had a significantly higher AUC compared with MELD (0.80 vs. 0.766, P<0.001). A MESO index <1.6 identified 97% of patients who survived at 3 months and the predicted survival rate was 96.5%. In survival analysis, MESO index >1.6 independently predicted a higher mortality rate (relative risk: 3.32, P<0001) using the Cox model. CONCLUSIONS: The MESO index, which takes into account the predictive power of both MELD and SNa, is a useful prognostic predictor for both short- and long-term survival in cirrhotic patients. PMID- 17403191 TI - Consequences of treatment withdrawal in type 1 autoimmune hepatitis. AB - BACKGROUND AND AIMS: Drug-related side effects are considered the major consequences of relapse and re-treatment in patients with autoimmune hepatitis. Our goals were to determine whether relapse is associated with disease progression and whether treatment end points can be refined. METHODS: The outcomes of 132 patients with definite type 1 autoimmune hepatitis who had been treated comparably until remission were assessed retrospectively after drug withdrawal. RESULTS: Patients who had relapsed repeatedly after initial treatment withdrawal developed cirrhosis more commonly than patients who sustained remission (18/48 vs 1/22, P=0.004), and those who relapsed once (18/48 vs 2/21, P=0.02). Hepatic death or the need for liver transplantation was also more frequent in the patients who had multiple relapses than those who sustained remission (13/64 vs 0/30, P=0.008) and those who relapsed once (13/64 vs 1/38, P=0.02). Patients who sustained their remission had a higher frequency of normal laboratory indices at drug withdrawal than patients who relapsed (88% vs 46%, P=0.003). Adverse outcomes after relapse did not distinguish patients until after 5 years of observation. CONCLUSIONS: Multiple relapses are associated with a poorer prognosis than sustained remission or single relapse episodes. Initial treatment to resolution of laboratory abnormalities may afford the greatest opportunity to prevent relapse. PMID- 17403192 TI - Dynamic MRI signals in the second week of radiotherapy relate to treatment outcomes of hepatocellular carcinoma: a preliminary result. AB - AIM: Radiotherapy (RT) has been used to treat hepatocellular carcinoma (HCC) in recent years. Despite its good local control, slow tumoral shrinkage and rapid recurrence compromise treatment outcomes. We evaluated the signal intensity of the hepatic parenchyma and tumours by using dynamic contrast enhanced magnetic resonance imaging (MRI) and correlated the findings with clinical outcomes. Nineteen patients with advanced HCC received 50 Gy in 25 fractions. They underwent a dynamic contrast-enhanced, turbo fast low-angle shot MR sequence at 1.5 T before therapy, at 2 weeks of therapy, and 1 month (week 9) later. Initial first-pass enhancement slopes (slope) and peak enhancement ratios (peak) were measured. RESULTS: Initial signal intensities were not associated with RT outcomes. An increased slope and peak of the tumour at week 2 was associated with an improved local response (P<0.05). In the parenchyma, an increased slope at week 2 was associated with recurrence outside the radiation fields or with progression over distant sites (P<0.05). The differences in signal changes at week 2 during RT were not persistent at a statistically significant level at 1 month after RT. CONCLUSION: Dynamic contrast-enhanced MRI signals may act as biomarkers for early prediction of responses to RT in patients with HCC. Signal intensities at week 2 are important in evaluating treatment outcomes. PMID- 17403193 TI - Apocynin alleviated hepatic oxidative burden and reduced liver injury in hypercholesterolaemia. AB - BACKGROUND: This study addressed the effects of apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, on hepatic oxidative burden and liver injury during diet-induced hypercholesterolaemia. METHODS AND RESULTS: Male Wistar rats were fed a 4% cholesterol-enriched diet for 3 weeks. Apocynin was administered in drinking water concurrently. The high-cholesterol diet (HC) significantly increased the serum level of cholesterol and hepatic cholesterol ester deposition, and these parameters were similar between the HC and high cholesterol diet plus apocynin (HCA) groups. The HC group showed abnormal liver function tests [alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (Alk-P)] as well as increased Evans blue extravasation and macrophages infiltration. Apocynin treatment could suppress these inflammation-related parameters. In vivo measurement of NADPH-derived cellular autofluorescence suggested that HC increased oxidative stress in hepatocytes. Biochemical analysis of redox status including thiobarbituric acid reactive substances, reduced glutathione, and oxidized glutathione also confirmed the phenomenon. Apocynin treatment was able to alleviate these indices of oxidative burden owing to HC. Furthermore, apocynin-abrogated HC induced gp91(phox) expression, suggesting the involvement of NADPH oxidase in the pathogenesis. CONCLUSIONS: We concluded that apocynin suppressed NADPH oxidase activation and subsequent liver injuries owing to high-cholesterol intake in rats. The impacts of cholesterol metabolism disorders on pathogenesis and progression of steatohepatitis warrant further clinical investigation. PMID- 17403194 TI - Endogenous opioids modulate hepatocyte apoptosis in a rat model of chronic cholestasis: the role of oxidative stress. AB - AIMS/BACKGROUND: There are increasing number of evidences indicating the contribution of endogenous opioids in the pathophysiology of cholestatic liver disease. The aim of the present study was to determine the role of the endogenous opioid system in the modulation of hepatocytes apoptosis and liver oxidant/anti oxidant balance during chronic cholestasis in rats. METHODS: We induced cholestasis in rats by bile duct ligation (BDL). Naltrexone, an opioid antagonist, was administered at different doses (2.5, 5, 10, 20 and 40 mg/kg/day) to cholestatic animals for 5 weeks. RESULTS: Naltrexone prevented the cholestasis induced decrease of hepatic glutathione levels at higher doses (20 and 40 mg/kg/day). In the next phase of the study, we evaluated the effects of 20 mg/kg/day naltrexone treatment on hepatic damage indices and liver oxidant/anti oxidant balance in 5-week BDL rats. There was a marked increase in the number of apoptotic hepatocytes as well as serum liver enzymes and hepatic lipid peroxidation levels in cholestatic rats compared with sham-operated animals 5 weeks after the operation. Liver anti-oxidant enzyme activities were significantly reduced in cholestatic rats compared with controls. Chronic treatment with naltrexone significantly improved all the aforementioned indices in comparison with saline-treated cholestatic rats. CONCLUSION: Our findings demonstrate that the administration of opioid antagonist is protective against hepatic damage in a rat model of chronic cholestasis. We suggest that increased levels of endogenous opioids contribute to hepatocytes apoptosis in cholestasis, possibly through downregulation of liver anti-oxidant defense. PMID- 17403195 TI - Hepatic stellate cell-targeted delivery of M6P-HSA-glycyrrhetinic acid attenuates hepatic fibrogenesis in a bile duct ligation rat model. AB - BACKGROUND/AIMS: Hepatic stellate cells (HSCs) play a key role in fibrogenesis. Here, we used mannose-6-phosphate-modified human serum albumin (M6P(26)-HSA) as a selective carrier to deliver antifibrotic drug 18beta-glycyrrhetinic acid (18beta GA) in experimental fibrosis animals, and tested its effect in injured liver tissues. METHODS: Bile duct ligation (BDL) was performed to induce liver damage in rats. Masson's stain and immunocytochemistry were used to assess hepatic collagen deposits and uptakes of M6P(26)-HSA-GA in HSCs in rat livers. Gene expression profiles of procollagen type I alpha2, smooth muscle actin (SMA), and transforming growth factor-beta1 (TGF-beta1) were analysed by TaqMan and quantitative polymerase chain reaction assays. The depositions of M6P(26)-HSA-GA in the HSC-T6 cell line and primary HSCs were assessed by immunofluorescent staining. RESULTS: Treatment with M6P(26)-HSA-GA at 10 mg/kg (three times/week for 2 weeks), but not the equivalent doses of free 18beta-GA and M6P(26)-HSA carrier alone, could significantly attenuate collagen deposits in BDL rat liver. Masson's stain and TaqMan assay revealed significant modulation of procollagen type I alpha2 in the BDL-injured liver. The depositions of M6P(26)-HSA-GA in HSCs were revealed by immunostaining with HSA and SMA markers. M6P(26)-HSA bound activated HSCs in vitro and the immunoreactivity of M6P(26)-HSA-GA was detected in the cytoplasm and cell surface of HSCs and HSC-T6 cells. The gene transcript levels of SMA and TGF-beta1 were modulated in HSC-T6 cells treated with M6P(26) HSA-GA. CONCLUSIONS: The M6P(26)-HSA holds promise as a targeting carrier for the liver or HSCs, which may be used to deliver 18beta-GA as a therapeutic agent to treat liver fibrosis. PMID- 17403196 TI - Cytoprotective effects of taurocholic acid feeding on the biliary tree after adrenergic denervation of the liver. AB - BACKGROUND: Cholangiopathies impair the balance between proliferation and apoptosis of cholangiocytes leading to the disappearance of bile ducts and liver failure. Taurocholic acid (TC) is essential for cholangiocyte proliferative and functional response to cholestasis. Bile acids and neurotransmitters co operatively regulate the biological response of the biliary epithelium to cholestasis. Adrenergic denervation of the liver during cholestasis results in the damage of bile ducts. AIM: To verify whether TC feeding prevents the damage of the biliary tree induced by adrenergic denervation in the course of cholestasis. METHODS: Rats subjected to bile duct ligation (BDL) and to adrenergic denervation were fed a TC-enriched diet, in the absence or presence of daily administration of the phosphatidyl-inositol-3-kinase (PI3K) inhibitor wortmannin for 1 week. RESULTS: TC prevented the induction of cholangiocyte apoptosis induced by adrenergic denervation. TC also restored cholangiocyte proliferation and functional activity, reduced after adrenergic denervation. TC prevented AKT dephosphorylation induced by adrenergic denervation. The cytoprotective effects of TC were abolished by the simultaneous administration of wortmannin. SUMMARY/CONCLUSIONS: TC administration prevents the damage of the biliary tree induced by the adrenergic denervation of the liver. These novel findings open novel perspectives in the understanding of the potential of bile acids especially in post-transplant liver disease. PMID- 17403197 TI - Adenovirus-mediated hepatic syndecan-1 overexpression induces hepatocyte proliferation and hyperlipidaemia in mice. AB - BACKGROUND: Heparan sulfate proteoglycans (HSPGs) have been involved in the regulation of cell growth, apoptosis and lipid metabolism in vitro; however, their functional role in vivo remains unknown. AIM: Here, we describe hepatic tissue and lipid metabolism changes after liver overexpression of syndecan-1 (SDC 1), the main hepatic HSPG, in mice induced by adenoviral gene transfer. RESULTS: SDC-1 overexpression was associated with marked hepatocyte proliferation, cell isolated apoptosis and increased plasma alanine aminotransferase (ALT) levels. Additionally, SDC-1 liver overexpression significantly raised plasma cholesterol and triglyceride concentrations due to an increase in all lipoprotein particles, including the appearance of large and apolipoprotein (apo) E-enriched high density lipoprotein (HDL) particles. Hepatic very low-density lipoprotein (VLDL) production was not affected by SDC-1 overexpression, suggesting a delayed plasma clearance of apo B lipoproteins as the underlying hyperlipidaemic mechanism. These pleotropic effects were qualitatively equivalent, even though less intense, in mice overexpressing a cytoplasmic C-terminal domain-deleted SDC-1. CONCLUSIONS: This is the first report in vivo of the biological effects induced by a specific HSPG in the liver, with potential implications in both regenerative biology and molecular lipidology. PMID- 17403198 TI - Baicalin protects mouse from Concanavalin A-induced liver injury through inhibition of cytokine production and hepatocyte apoptosis. AB - BACKGROUND: Baicalin (BA) exhibits an anti-inflammatory effect in vivo and in vitro and is used to treat chronic hepatitis. However, the mechanism by which BA exerts the liver-protective effect remains largely unknown. AIMS: The present study reports that BA inhibits cytokine production and hepatocyte apoptosis to protect mice from liver injury induced by concanavalin A (Con A), a T-cell dependent liver injury model. RESULTS: Con A injection of mice induced severe immune responses and extensive hepatocellular apoptosis within 24 h. Pretreatment of 200 or 100 mg/kg BA markedly reduced serum aminotransferase activities, protected hepatoycte apoptosis and reduced the increase of plasma cytokine levels, including tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma) and interleukin-6 (IL-6). Furthermore, BA pretreatment decreased tissue myeloperoxidase activity and lipid peroxidation, but increased the superoxide dismutase level. In vitro studies indicated that the beneficial effect of BA was associated with reduced cytokine production from lymphocytes and reduced TNF alpha-induced hepatocyte apoptosis. CONCLUSION: These results suggest that BA has therapeutic potential for T-cell-mediated liver injury. PMID- 17403199 TI - Autoimmune hepatitis after treatment with fluvastatin. PMID- 17403200 TI - A guide to murine platelet structure, function, assays, and genetic alterations. AB - Platelets play an important role in hemostasis, thrombosis and several other biological processes. The adaptability of mice to genetic manipulation and their genetic similarity to humans has resulted in a plethora of murine models to study platelet function. Although murine platelets differ from human platelets with regard to size, number and structure, functionally they are very similar. Thus, studies which employed these model systems have greatly improved our current understanding of the contribution of platelets to hemostasis and thrombosis. This review presents general recommendations with respect to collection, isolation and processing of murine platelets. It also describes the assays currently available to study platelet function and critically assesses their utility. The extensive literature on the effects of genetic alterations on murine platelet function is considered in detail. This review is intended to provide a convenient source of reference for platelet investigators. PMID- 17403201 TI - A guide to murine coagulation factor structure, function, assays, and genetic alterations. AB - Murine blood coagulation factors and function are quite similar to those of humans. Because of this similarity and the adaptability of mice to genetic manipulation, murine coagulation factors and inhibitors have been extensively studied. These studies have provided significant insights into human hemostasis. They have also provided useful experimental models for evaluation of the pathophysiology and treatment of thrombosis. This review contains recommendations for obtaining, processing and assaying mouse blood hemostatic components, and it summarizes the extensive literature on murine coagulation factor structure and function, assays and genetic alteration. It is intended to be a convenient reference source for investigators of hemostasis and thrombosis. PMID- 17403202 TI - A guide to murine fibrinolytic factor structure, function, assays, and genetic alterations. AB - The components and functions of the murine fibrinolytic system are quite similar to those of humans. Because of these similarities and the adaptability of mice to genetic manipulation, murine fibrinolysis has been studied extensively. These studies have yielded important information regarding the function of the several components of fibrinolysis. This review presents information on the structure, function and assay of mouse fibrinolytic parameters and it discusses the results of the extensive studies of genetically modified mice. It is intended to be a convenient reference resource for investigators of fibrinolysis. PMID- 17403203 TI - Release of biologically active CD154 during collection and storage of platelet concentrates prepared for transfusion. AB - BACKGROUND: Millions of platelet transfusions are given each year. Transfusion reactions occur in as many as 30% of patients receiving unmodified platelet transfusions. The cause of some transfusion reactions remains unclear. The current paradigm suggests that platelet concentrates (PC) contain proinflammatory mediators that are released by white blood cells during collection, processing and storage. CD154 (CD40 ligand, CD40L) is a potent inflammatory mediator, normally sequestered inside the resting platelet, that is known to translocate to the platelet membrane and be shed into plasma in response to agonist activation. We hypothesized that platelet-soluble CD154 (sCD154) is 'spontaneously' released by transfused platelets and plays a major role in transfusion reactions. OBJECTIVES: To determine the time course and biological properties of CD154 translocation and release during collection and storage of platelets for transfusion. METHODS: We measured surface and sCD154 in platelets prepared by the platelet-rich plasma method or apheresis by fluorescence-activated cell sorting and enzyme-linked immunosorbent assay, respectively. The specific biological activity of platelet sCD154 was assayed by stimulation of the CD154/CD40 pathway in known CD40-positive cells with PC-derived supernatants. RESULTS AND CONCLUSIONS: We demonstrate that PCs prepared for transfusion have high levels of membrane-bound CD154 and sCD154, with maximum levels being seen 72 h after platelet collection. Importantly, we show that platelet-derived sCD154 potently stimulates CD40-positive cells. We propose that platelet-derived CD154 is a key 'cytokine' responsible for adverse reactions associated with platelet transfusions. Improved methods of platelet collection and/or storage, which limit CD154 expression, could reduce the risks of transfusion reaction. PMID- 17403204 TI - JAK2 V617F is specifically associated with idiopathic splanchnic vein thrombosis. PMID- 17403205 TI - Standardization of circulating endothelial cell enumeration by the use of human umbilical vein endothelial cells. PMID- 17403206 TI - Ticlopidine could be an alternative therapy in the case of pharmacological resistance to clopidogrel: a report of three cases. PMID- 17403207 TI - Evaluation of the platelet response to clopidogrel with light transmittance aggregometry: peak aggregation or late aggregation? PMID- 17403210 TI - There is no 'truth' outside a context: implications for the teaching of analytical psychology in the 21st century. AB - Humans are from birth embedded in a historical and contemporary context of meanings. This always constrains their theoretical and practical activities. In this paper, I will be suggesting that there are no guiding 'truths' outside such contexts. In order to understand the foundations of any concept or new idea, it is important to comprehend the context in which it is embedded. Candidates and some of their teachers have very little knowledge of the intellectual context in which Jung or any other analytic theorist wrote. As a result, the analytic 'founders' are often believed to have discovered 'truths' transcending the context of history and of everyday life. They were, however, as much creative synthesizers as pure originators. I propose that the components of all analytic theories emerge from their embeddedness. I contend, therefore, that the understandings of these components such as psychic causality, epistemology and identity formation were quite differently conceived at the time the analytic founders were working than they are today. I will give examples to show how the acceptance of this attitude is useful in inculcating a discerning approach in candidates towards traditionally accepted analytic theories and practices. This can provide them with a method to promote their individual creative development. PMID- 17403211 TI - The alchemy of training. AB - Training in Zurich has its own special character but also is marked by the very fact that it is in Zurich. Zurich radiates its own distinct energy and carries a specific historical significance in the world of analytical psychology. This, like all things with psychic energy, has a spectrum of meaning. This spectrum, as well as the 'spirit' of the place, will be critically examined, taking into account the 'blessings' and 'curses' of such genius loci. Training in Zurich is experientially based and is first and foremost an initiation: an initiation into symbolic life, or rather life where symbol plays an important role. Training is understood to involve a transformation of one's self, much like the 8th century alchemist Morienus Romanus understood the opus as a 'human transformation system'. It is not merely an education. The requirement of 'immersion' is core to the experience of becoming an analyst in Zurich and this sets up a valuable discomfort between rational intellectual learning and intuitive experience, between knowing and not-knowing. How does this dis-serve the making of an analyst? What is implicit in this immersion and its discomfort? Does it have a role in today's emphasis on clinical and empirical training? Does Zurich still offer something unique and valuable in the world of training, or is it passe? From these questions, the dichotomy of what is 'urgent' and 'essential' in training will be examined. PMID- 17403212 TI - The making of an analyst: from 'ideal' to 'good-enough'. AB - This paper addresses the issue of variances in training modalities and how this is linked to one's personal experience of training and to one's analytic lineage. The author, who bases his reflections on discussions held during the yearly directors of training meeting of the North American Jungian Societies, suggests that, while each institute aims to provide an 'ideal' training programme for its candidates, the philosophy underlying how this ideal is defined depends, in large part, on the theoretical and philosophical orientation of the founding fathers and mothers. This results in a form of analytic lineage that necessarily impacts on the form and content of the 'ideal' programme. Shadow issues related to analytic lineage in the admissions procedures, case consultation, exam committees and review committees are presented. Motivation for why we choose to train and reflections on what makes an analyst 'Jungian' are explored. PMID- 17403213 TI - Evaluating progress in training: character or competence? AB - Jung was mercurial in his attitudes to method and technique, leaving us a problematic legacy when it comes to evaluating the progress of trainees. Some would say that those of us involved with the assessment of candidates during their training continue to rely too heavily on intuition and subjective assessments. However good our admissions' procedures and the structures in place to review progress, the emotional and financial demands for trainees of embarking on an analytic training, the tendency for analytic institutes to remain opaque and slow to link up with the external world and the cliques within our profession make more objective assessments of progress and readiness to qualify at best haphazard and at worst inadequate. Some trainees have an immediate talent for analytic work; others develop their gifts more slowly; some never find this capacity. Working from a definition of analytic talent, the paper begins to map out a Jungian framework for assessing progress, emphasizing the significance of both character and competence and the developing relationship between them. PMID- 17403214 TI - Fordham, feeling, and countertransference: reflections on defences of the self. AB - This is a paper about the difficulties we as analysts get into when we find that a patient has activated something in our unconscious which we cannot resolve in our work with them. Fordham described at the end of his life, in a number of papers, his difficulties and discomfort at not being able to resolve an impasse with one of his patients. From the conversations we had about this situation I knew this caused them both a lot of pain. After Fordham's death his former patient consulted me. Arising from these consultations I describe how I have understood the impasse to have arisen between Fordham and his patient. This paper links character and clinical interests, personality and impasse, developmental failures and defences of the self. It is a personal statement in which I have struggled to represent the meaning in the pain these two men suffered during their analytic engagement, which lasted more than ten years. The theme of fathers and sons was central to the problem. PMID- 17403215 TI - 'What works?' Response to the paper by James Astor. AB - A personal account of an analysis with Michael Fordham and subsequent conversations with James Astor. A number of themes are developed concerning the expression of feeling, use of language, and the relevance of aesthetic values to the practice and reporting of analysis. PMID- 17403216 TI - Response to Astor's paper. PMID- 17403222 TI - Education qualification levels and school careers of unilateral versus bilateral hearing aid users. AB - OBJECTIVE: Analyse the difference in school careers and secondary school qualification levels between unilateral hearing aid users and bilateral hearing aid users. STUDY DESIGN: Retrospective questionnaire study. SETTING: Postal-based questionnaire. PARTICIPANTS: Names of adults known to have been fitted with unilateral or bilateral hearing aids during childhood were retrieved. This resulted in 292 names. Participants were selected using the following criteria: availability of the medical record, presence of bilateral hearing loss, completed secondary school education, normal IQ and a minimum aided word-recognition score of 70% at 10 years of age. The questionnaire was sent to 50 potential participants of whom 40 responded, resulting in two groups comprising 19 unilateral and 21 bilateral hearing aid users. METHODS: A questionnaire was developed to obtain data on secondary school levels and aspects of the school careers, such as type of schools attended (mainstream versus special), repeating a school year and additional assistance (speech and language therapy, personal frequency modulation systems). MAIN OUTCOME MEASURES: Comparison of the benefit of bilateral hearing aid fitting versus unilateral hearing aid fitting on secondary school qualification level, type of school the children attended, class failure and additional assistance used. RESULTS: The group of bilaterally fitted hearing aid users eventually completed secondary school at a superior level than the unilaterally fitted users. 33% of binaural hearing aid users achieved qualifications giving access to a bachelors degree compared with 21% of unilateral users. There were no differences between those fitted with unilateral and bilateral hearing aids in the type of schools attended, repeating a school year and additional assistance. CONCLUSIONS: The bilateral hearing aid users obtained superior secondary school qualifications than the unilateral users but their school careers were comparable. PMID- 17403223 TI - Analysis of paranasal sinus development and anatomical variations: a CT genetic study in twins. AB - OBJECTIVE: To determine if anatomical variations in the paranasal sinuses of twins are the result of genetic or environmental influences. STUDY DESIGN: A prospective, observational study. SETTING: An academic hospital in Brussels, Belgium. PARTICIPANTS: Twenty-five volunteer twins (17 identical twins and eight non-identical twins). Of the identical twins, six were both female and 11 both male. The mean age was 40.6 years (18-49). Of the non-identical twins, one pair was female and seven were male. The mean age was 39.6 years (25-48). MAIN OUTCOME MEASURES: The CT-scan similarity in anatomical structures (frontal cells, agger nasi cells, concha bullosa, infraorbital cells and changes in the shape of the lamina orbitalis), and the intranasal and paranasal sinus morphology was compared between the twins. RESULTS: Among the studied variables, there was no statistically significant difference between the two twin groups (Fisher's exact test, P > 0.05). Regarding combine-studied parameters, there were no identical CT scans in the twins. However, a tendency of non-genetic influence existed in the development of frontal cell type III and IV, and infraorbital cells, as well as a liability of genetic influence in the presence of concha bullosa. CONCLUSION: In this study, the differences in anatomical structure of the paranasal sinuses between identical and non-identical twin pairs were not statistically significant. This indicates that environmental factors are more significant than genetic ones in the development of anatomical variations in paranasal sinus anatomy. PMID- 17403224 TI - Quinsy treated by aspiration: the volume of pus at initial aspiration is an accurate predictor of the need for subsequent re-aspiration. AB - OBJECTIVE: The aim of this study was to determine an accurate indicator of the need for second aspiration of peritonsillar abscesses the day after initial aspiration. SETTING: A tertiary otolaryngology care centre. PARTICIPANTS: Fifty patients aged between 11 and 49 years with suspected peritonsillar abscess. STUDY DESIGN: A prospective case series. OUTCOME MEASURES: The potential indicators investigated included volume of pus at initial aspiration and clinical indicators suggesting persistent pus (dysphagia, odynophagia and trismus). The outcome measure was the presence of pus at subsequent aspiration. RESULTS: A linear correlation was found between volume of first aspirate and presence of pus on re aspiration (r = 0.9753). A volume of pus <3 mL on initial aspiration accurately predicted <0.5 mL pus on re-aspiration. Sixty-four per cent (32) patients had 3 mL or more pus on initial aspiration and in all there was at least 1 mL or more pus on second aspiration. Clinical indicators correlated less well, with a average coefficient on first aspiration of 0.62 and on second aspiration of 0.35. CONCLUSION: The volume of pus on initial aspiration is a very reliable indicator in assessing the need for re-aspiration of peritonsillar abscesses. If 3 mL or more of pus are aspirated on the first occasion these patients should be seen the next day and have a further aspiration. Clinical symptoms and signs are not useful indicators. PMID- 17403225 TI - Does the addition of sublingual gland excision to submandibular duct relocation give better overall results in drooling control? AB - Submandibular duct relocation plus or minus excision of the sublingual glands are relatively simple procedures with low morbidity. Between 1981 and 2005, 71 submandibular duct relocation and 29 submandibular duct relocation plus excision of the sublingual glands procedures were conducted. OBJECTIVES: To compare both procedures including operative time, length of hospital stay, postoperative complications, drooling scores and parental satisfaction. STUDY DESIGN: Prospective study. SETTING: Paediatric tertiary referral centre. PARTICIPANTS: Patients referred with excessive drooling after failure of conservative methods. EXCLUSION CRITERIA: patients with recurrent aspiration pneumonias or dental caries. Two patients were lost to follow up and excluded from the study. RESULTS: Operative time and length of hospital stay were increased in the submandibular duct relocation plus sublingual gland excision group. Drooling scores and parental satisfaction results were excellent, 93% of parents in the submandibular duct relocation group and 89% of parents in the duct relocation plus sublingual glands excision were satisfied and would recommend the procedure. There was no statistical difference (P = 0.643) in drooling scores between the two procedures. Postoperative morbidity was higher with the addition of sublingual gland excision, with postoperative haemorrhage occurring in 13.7% and 36% of parents expressing concern over postoperative pain, compared with 3% postoperative haemorrhage rate with submandibular duct relocation and only 12% of parents expressing the same concerns. CONCLUSION: We conclude that both procedures are effective in drooling control, but the addition of sublingual gland excision increases morbidity and we are no longer excising these glands with submandibular duct relocation. PMID- 17403226 TI - Lump in the throat. PMID- 17403230 TI - The modified 'Parent's Kiss' for the removal of paediatric nasal foreign bodies. AB - Paediatric nasal foreign bodies is a common clinical problem effecting children mainly 2-8 years of age. Often treated in A & E Departments, with little or no specialised ENT experience or equipment. We present a simple, parent administered technique for removal of paediatric nasal foreign bodies. The technique is safe and effective and is easy to perform on nervous and frightened children. PMID- 17403231 TI - 'Boil and Bite' mouth guards for direct laryngoscopy. AB - 'Boil and Bite' mouth guards are an alternative to conventional silicone guards in direct laryngoscopy. These guards are less likely to displace. They are cheap and easy to prepare. PMID- 17403232 TI - Comparing two models of voice care delivery. AB - Voice care is an important aspect in the successful treatment of patients with non-organic dysphonia. No previous study has examined the specific effect of voice care programmes or the comparative effectiveness of the delivery of these programmes across two healthcare professionals (speech and language therapists and ENT nurses). We describe a pilot study that provides preliminary data that suggests a short, easily deliverable voice care programme is (i) effective in reducing patients' perception of their dysphonia and (ii) equally effectively administered by an ENT nurse compared with a speech and language therapist. The study provides a firm base for continued research. PMID- 17403233 TI - Intraparotid facial nerve schwannoma. What to do? AB - An intraparotid facial nerve schwannoma is often not recognised in pre-treatment work-up and frequently results in subsequent significant postoperative morbidity. We have evaluated the literature regarding pre-treatment work-up and facial nerve function outcome. Two of our own cases are presented. A minority of the intraparotid schwannomas can be removed by resection while preserving facial nerve integrity and function. In the event of preoperative facial nerve dysfunction, tumour resection and subsequent nerve repair should be considered. If resection of an intraparotid facial nerve schwannoma cannot be performed with preservation of facial nerve integrity and function, a wait-and-see policy seems justified due to the indolent behaviour of the tumour and moderate results of facial nerve reconstruction. PMID- 17403234 TI - Teleconferencing in the management of head and neck cancer. AB - Teleconferencing can be used as part of the integrated management of head and neck cancer. An audit of the participants' views would suggest that the meeting is used in the management of complex head and neck issues. PMID- 17403235 TI - Pilot study of the knowledge and attitude towards evidence based medicine of otolaryngology higher surgical trainees. AB - The objective of this survey is to provide an insight into the knowledge and attitude of the higher surgical trainees in Otolaryngology towards evidence based medicine and evaluate their understanding of medical statistics and use of evidence based medicine in patient management. A self administered anonymous questionnaire was distributed to twenty Specialist Registrars in Otolaryngology in Ireland during an annual National Otolaryngology Meeting. Of these, nineteen attempted the questionnaire and eighteen completed it. Three previously published questionnaires [Meserve et al. (2005) Med. Educ. Online.10, 1; McColl et al. (1998) BMJ.316, 361; Khan et al. (1999) J. Obst. Gynaecol.19, 231] were used in this survey. In general, there was a moderately welcoming attitude towards evidence based medicine. However, the survey findings indicate the need for incorporation of formal teaching of critical appraisal skills into medical education with a greater emphasis on involvement and dissemination of evidence based medicine in day to day practice. PMID- 17403236 TI - Topical adrenaline in control of intraoperative bleeding in adenoidectomy: a randomised control trial. PMID- 17403238 TI - A negative but positive Cochrane review. PMID- 17403237 TI - Topical adrenaline in control of intraoperative bleeding in adenoidectomy: a randomised control trial. PMID- 17403239 TI - Indirect laryngoscopy - the cold way. PMID- 17403240 TI - Use of a towel clip to prevent slipping of Draffin rods. PMID- 17403241 TI - Demystifying the technique of aspirating a quinsy. PMID- 17403242 TI - Removal of an impalpable thyroglossal cyst using ultrasound-guided wire localisation - a technical note. PMID- 17403243 TI - How to take a look in the oesophagus. PMID- 17403244 TI - Nottingham tonsillectomy haemostasis simulator. PMID- 17403245 TI - The benefits of ultrasound clinics alongside head and neck one-stop clinics. PMID- 17403246 TI - The FY2 tonsillar tie trainer. PMID- 17403247 TI - In suspense: an easy way to prevent contamination of the Zoellner sucker. PMID- 17403248 TI - Use of face mask in the management of epistaxis. PMID- 17403249 TI - A new technique for monopolar haemostasis in inferior turbinate surgery. PMID- 17403250 TI - Scoring systems, the Freedom of Information Act and selection to higher surgical training posts in Otolaryngology. PMID- 17403251 TI - Stuart Gatehouse. PMID- 17403254 TI - Nail disorders--one of dermatology's last frontiers. PMID- 17403255 TI - Dermoscopy provides useful information for the management of melanonychia striata. AB - The diagnosis of melanonychia striata is often difficult, and a biopsy of the nail matrix is required in doubtful cases. However, dermoscopic examination of the nail plate offers interesting information in order to better select the cases in which pathologic examination is indicated. In the case of brown longitudinal pigmentation with parallel regular lines, the diagnosis of nail apparatus melanocytic nevus could be made. On the other hand, the presence of a brown pigmentation overlaid by longitudinal lines irregular in their thickness, spacing, color, or parallelism is highly in favor of a melanoma. Gray homogeneous lines are observed in case of lentigo, lentiginoses, ethnic or drug-induced pigmentations, and in post-traumatic pigmentations. Blood spots are characterized by their round-shaped proximal edge and their filamentous distal edge and are highly suggestive of subungual hemorrhages. Dermoscopic examination of the free edge of the nail plate gives information on the lesion location; pigmentation of the dorsum of the nail plate is in favor of a proximal nail matrix lesion, whereas pigmentation the lower part of the nail edge is in favor of a lesion of the distal matrix. PMID- 17403256 TI - Clinical pathologic correlations for diagnosis and treatment of nail disorders. AB - Clinicopathologic correlation is crucial to the correct diagnosis of disorders of the nail unit. This chapter will explore four common clinical scenarios and how pathology can help differentiate between their various etiologies. These include: dark spot on the nail plate (melanin versus heme), subungual hyperkeratosis (onychomycosis versus psoriasis), longitudinal melanonychia (benign versus malignant), and verrucous papule (verruca versus squamous cell carcinoma). Consideration must be given to both when to perform a biopsy and the location of the biopsy site, which must be based on an understanding of the origin of the changes. An overarching principle is that lesions within the same differential diagnosis may be present concomitantly, such as malignant melanoma of the nail unit associated with hemorrhage. Therefore, even with a biopsy-proven diagnosis, the clinician must always monitor lesions of the nail unit for appropriate response to treatment and consider an additional biopsy for recalcitrant lesions. PMID- 17403257 TI - Autoimmune disorders: nail signs and therapeutic approaches. AB - Systemic sclerosis (scleroderma, SSc) is an autoimmune disease that targets small and medium-sized arteries and arterioles in the involved tissues, resulting in a fibrotic vasculopathy and tissue fibrosis. Several prominent nail and periungual changes are apparent in scleroderma. Examination of the nail fold capillaries can reveal the nature and extent of microvascular pathology in patients with collagen vascular disease and Raynaud's phenomenon. Among the complications stemming from Raynaud's phenomenon can be painful ischemic digital ulcers. This can be managed, and potentially prevented, through pharmacologic and nonpharmacologic means. Whereas oral calcium channel blockers remain the most convenient therapy, oral endothelin receptor antagonists and intravenous prostaglandins may be important therapeutic advances for ischemic digital vascular lesions. PMID- 17403258 TI - Toenail onychomycosis: current and future treatment options. AB - Onychomycosis is a common disease affecting as much as 8% of the general population. Treatment of onychomycosis is challenging, complicated by low cure rates and relatively high relapse rates. This paper reviews the efficacy of current oral, topical, and surgical treatment options. Currently, the treatment of choice for toenail onychomycosis is oral terbinafine because of its high efficacy, low relapse rates, and cost-effectiveness. Oral itraconazole or fluconazole could be considered for infections caused by Candida. Topical therapies may be a useful adjunct to these systemic therapies, but are less effective when used alone. More research is needed to determine the best measures for preventing reinfection. PMID- 17403259 TI - Contact and primary irritant dermatitis of the nail unit diagnosis and treatment. AB - Contact dermatitis of the nail unit is not rare. The most common allergens are found in nail cosmetics such as enamel, sculptured nails, and preformed plastic tips. Toluene sulfonamide formaldehyde resin, acrylates, and ethylcyanoacrylate are the most common allergens. Along with a proper clinical history focusing on nail practices, a thorough understanding of the common allergens and patch testing are necessary to diagnose contact dermatitis. A discussion of the common allergens and irritants relevant to the nail unit will be presented. PMID- 17403260 TI - Treating cosmetically induced nail problems. AB - Smooth, lustrous nails are a sign of health and beauty in our society and fuel the US$6-billion nail salon industry in the United States. Although many women can use nail cosmetics without adverse consequences, when problems occur it is important to recognize the causes and treat the problem condition. The cornerstone of management of nail cosmetic problems is prevention through education. In spite of efforts to that end, nail salon procedures and materials can cause nail disease that must be recognized and treated in order to restore the nails to health. PMID- 17403261 TI - Biological therapy and nail psoriasis. AB - Nail psoriasis affects 50% of psoriasis patients and in many cases causes impairment of manual dexterity, pain, and psychologic stress. Despite the fact that about 80% of psoriatic arthritis patients have nail involvement, patients rarely receive treatment for nail disease. Because of great difficulty in drug delivery to site of action and significant toxicities of most conventional systemic therapies, treatment of nail psoriasis can be very challenging. Biological therapy for psoriasis and psoriatic arthritis is now widely used, and early evidence indicates it may have significant benefit for some patients with psoriatic nail disease. This article reviews the clinical manifestations of nail psoriasis and discusses how to use specific biologic therapies that may provide significant new treatment options for this challenging disease. PMID- 17403262 TI - Nail surgery: practical tips and treatment options. AB - This paper provides an introductory overview of nail surgery, including basic principles, anesthesia, wound dressings, and several common nail procedures. In detail, it reviews how to achieve complete anesthesia and hemostasis, perform matrix injection, punch biopsy of the plate, bed, and matrix, and nail fold biopsy, with practical tips and suggested surgical indications. PMID- 17403263 TI - Accurate consent for insertion and later removal of grommets. AB - Each year in the UK over 30,000 patients undergo insertion of grommets. The grommet insertion may cause many problems like persistent otorrhoea, scarred drum, retraction pockets and retention. The grommets may be extruded from the middle ears by the normal epithelial migration mechanism once they have served their purpose. These may become infected and require removal. We have analysed the Department of Health Hospital Episode Statistics relating to the insertion and removal of grommets (ventilation tubes). We have shown that 7.6 per cent of patients who have grommets inserted will have grommets removed. PMID- 17403264 TI - Nurse-led 'one stop' clinic for elective tonsillectomy referrals. AB - OBJECTIVES: Balancing new referrals with limited clinic capacity is a challenge. At Lincoln County Hospital, referrals for tonsillectomy have been managed by an experienced ENT nurse practitioner, in order to deal with this problem more effectively. We reviewed tonsillectomy referrals made in a one year period to determine if surgical outcomes were satisfactory. Results were compared with figures available from the national prospective tonsillectomy audit. METHODS: This was a retrospective study, assessing patients referred to the ENT department at Lincoln County Hospital. We reviewed the number of patients who had proceeded to surgery and the rate of complications. RESULTS: One hundred and fifty-nine patients had been referred for tonsillectomy over the one year period. One hundred and forty-seven patients had been seen in clinic and 12 had not attended their clinic appointment. Following consultation with the nurse practitioner, 125 patients had proceeded to surgery. Five episodes of bleeding were recorded as complications postoperatively; giving a bleeding complication rate of 4.7 per cent, compared with a rate of 5.7 per cent from the national prospective tonsillectomy audit. CONCLUSION: A trained nurse practitioner can safely and effectively facilitate the complete process of managing a routine pre-operative tonsillectomy patient, from primary care referral to arranging admission for surgery. PMID- 17403265 TI - Management of tinnitus induced by brainstem and cerebellar infarction associated with complications of cerebello-pontine angle surgery. AB - Following surgery in the USA in 1992 to remove a large right cerebello-pontine angle tumour, a 39-year-old woman developed severe brainstem and cerebellar infarction. This left her with severe visual impairment and ataxia. She became able to communicate by means of an adapted finger-spelling alphabet. She had total hearing loss in the right ear and a mild to moderately severe sensorineural hearing loss in the left ear, and severe tinnitus heard throughout the head. Additionally, she experienced hypersensitivity to sound above normal conversational levels, which evoked a synaesthetic feeling of coldness across her upper torso. Previous linear analogue hearing aid fitting had not been beneficial for either hearing or tinnitus. Careful fitting of a digital hearing aid, together with tinnitus counselling, inhibited the patient's tinnitus to 25 per cent of its former intensity after a six month acclimatisation period, and improved communication. PMID- 17403266 TI - Maxillary intraosseous vascular malformation: a case report and literature review. AB - We present a rare case of maxillary intraosseous vascular malformation and review the nomenclature used by pathologists and clinicians in classifying vascular anomalies. We also discuss the literature and highlight potential management pitfalls. PMID- 17403267 TI - Pre-operative nutritional status does not alter the metabolic response to major gastrointestinal surgery in patients with oesophageal cancer. AB - Malnutrition is associated with an increased incidence of perioperative morbidity and mortality. To evaluate the effect of malnutrition on the metabolic and inflammatory response to surgery in patients with oesophageal cancer, we studied the effects of oesophagectomy in six patients with major (13.9 (se 1.3) %) weight loss and five patients with minor (0.7 (se 0.6) %) weight loss in the 6 months before to surgery. Rates of appearance (Ra) of glucose, glycerol, leucine and urea were determined by stable isotopically labelled tracer infusion before and after surgery. C-reactive protein was measured as an inflammation marker. BMI was lower in the patients with major weight loss than those with minor weight loss (20.3 (se 0.7) and 24.9 (se 1.5) kg/m2, P = 0.02). With the exception of greater glucose Ra in the major weight loss than minor weight loss subjects (11.1 (se 0.3) v. 9.5 (se 0.3) mumol/kg per min, P = 0.01), there were no differences in substrate kinetics before surgery between groups. Surgery increased glucose Ra, leucine Ra and urea Ra by 41, 24 and 58 %, respectively, in the total group. Changes in substrate kinetics in response to surgery were not different between patients with major and minor weight loss. Surgery increased C-reactive protein concentrations to a comparable extent in both groups. In conclusion, major upper gastrointestinal tract surgery in patients with oesophageal cancer elicits a catabolic response, characterized by increased inflammation, glucose production and protein breakdown. However, this catabolic response does not seem to be influenced by pre-operative nutritional status. PMID- 17403268 TI - A dietary pattern rich in olive oil and raw vegetables is associated with lower mortality in Italian elderly subjects. AB - Dietary habits play an important role in healthy ageing. We have investigated the role of dietary patterns on overall mortality in a large series of Italian elderly, recruited in five EPIC cohorts in Northern (Varese and Turin), Central (Florence) and Southern Italy (Naples and Ragusa).A total of 5611 subjects (72.6 % women) aged 60 years or older, enrolled in 1993-1998, were prospectively followed (median 6.2 years), with 152 deaths (98 women). Four major dietary patterns were identified by using an exploratory factor analysis based on dietary information collected at enrollment. The associations between these dietary patterns and overall mortality were evaluated by Cox models adjusted for potential confounders. The 'Olive Oil & Salad' pattern, characterised by a high consumption of olive oil, raw vegetables, soups and poultry, emerged as being inversely associated with overall mortality in both crude and adjusted models. After adjustment for gender, age and caloric intake, overall mortality was reduced by approximately 50 % in the highest quartile and a significant trend emerged (P = 0.008). This association persisted after adjusting for several additional confounders (hazard ratio (HR) 0.50; 95 % CI 0.29, 0.86; P for trend = 0.02). An association of the 'Pasta & Meat' pattern (characterised by pasta, tomato sauce, red meat, processed meat, added animal fat, white bread and wine) with increased overall mortality was also suggested, but only for the highest quartile in a multivariate model. Dietary recommendations aimed at the Italian elderly population should support a dietary pattern characterised by a high consumption of olive oil, raw vegetables and poultry. PMID- 17403269 TI - Phyto-oestrogens and risk of prostate cancer in Scottish men. AB - A population-based case-control study of diet, inherited susceptibility and prostate cancer was undertaken in the lowlands and central belt of Scotland to investigate the effect of phyto-oestrogen intake and serum concentrations on prostate cancer risk. A total of 433 cases and 483 controls aged 50-74 years were asked to complete a validated FFQ and provide a non-fasting blood sample. Multivariate logistic regression analysis found significant inverse associations with increased serum concentrations of enterolactone (adjusted OR 0.40, 95 % CI 0.22, 0.71] and with the consumption of soy foods (adjusted OR 0.52, 95 % CI 0.30, 0.91). However, no significant associations were observed for isoflavone intake or serum genistein, daidzein and equol. This study supports the hypotheses that soy foods and enterolactone metabolised from dietary lignans protect against prostate cancer in older Scottish men. PMID- 17403270 TI - Food chromium content, dietary chromium intake and related biological variables in French free-living elderly. AB - Trivalent chromium (Cr3+) is an essential trace element involved in insulin function. Cr deficiencies result in decreased insulin sensitivity, glucose intolerance and an increased risk of diabetes. Cr status decreases with age suggesting that the elderly may be at high risk of Cr deficiency. This study aimed to provide information about the Cr content of foods in France and the Cr intake in French free-living elderly. We measured the food Cr content and daily Cr intake of freely chosen diets for 3 d in twelve French free-living elderly people and their Cr excretion and plasma hormonal related variables, leptin, insulin and cortisol. Considering the relationship between insulin resistance and oxidative stress, we also determined plasma thiobarbituric acid reactive substance, thiol groups and total and reduced glutathione. Although these subjects had well-balanced diets, their daily Cr intakes did not reach the French recommendations. The low Cr intakes were due to the low Cr density of the foods. We found a negative correlation between Cr intakes and insulin, BMI and leptin. PMID- 17403271 TI - Amino acids and immune function. AB - A deficiency of dietary protein or amino acids has long been known to impair immune function and increase the susceptibility of animals and humans to infectious disease. However, only in the past 15 years have the underlying cellular and molecular mechanisms begun to unfold. Protein malnutrition reduces concentrations of most amino acids in plasma. Findings from recent studies indicate an important role for amino acids in immune responses by regulating: (1) the activation of T lymphocytes, B lymphocytes, natural killer cells and macrophages; (2) cellular redox state, gene expression and lymphocyte proliferation; and (3) the production of antibodies, cytokines and other cytotoxic substances. Increasing evidence shows that dietary supplementation of specific amino acids to animals and humans with malnutrition and infectious disease enhances the immune status, thereby reducing morbidity and mortality. Arginine, glutamine and cysteine precursors are the best prototypes. Because of a negative impact of imbalance and antagonism among amino acids on nutrient intake and utilisation, care should be exercised in developing effective strategies of enteral or parenteral provision for maximum health benefits. Such measures should be based on knowledge about the biochemistry and physiology of amino acids, their roles in immune responses, nutritional and pathological states of individuals and expected treatment outcomes. New knowledge about the metabolism of amino acids in leucocytes is critical for the development of effective means to prevent and treat immunodeficient diseases. These nutrients hold great promise in improving health and preventing infectious diseases in animals and humans. PMID- 17403272 TI - Serum carnitine, triglyceride and cholesterol profiles in Korean neonates. AB - This study evaluated carnitine and lipid status of fifty Korean newborns. Each subject was assigned to two groups: one according to body weight at birth and the other according to gestational age. Serum total, HDL- and LDL-cholesterol were significantly lower and triacylglycerols were significantly higher, by 14 %, in the low birth weight infant (LBWI, 1310-2490 g) group compared with the normal birth weight infant (NBWI, 2570-4420 g) group. Neither birth weight nor gestational age affected serum total carnitine concentrations. However, serum ASAC (acid-soluble acylcarnitine) concentrations were 43 % higher (P < 0.001) in the LBWI group compared with the NBWI group, and approximately twice as high (P < 0.05) in the 28-32 gestational age group compared with the other gestational age groups. NEC (non-esterified acyl carnitine) fractions were significantly higher in the NBWI and 28-32 week groups (P < 0.001 and P < 0.05); consequently serum acyl/NEC carnitine ratios were four times higher in the LBWI group compared with the NBWI group and 2-3 times higher in the 25-32 week age group compared with the more advanced gestational age groups. Urinary carnitine excretion, including the NEC fraction and total carnitine, was significantly higher (P < 0.001) for LBWI than for NBWI. By gestational age, NEC excretion of the 28-32 week group was significantly (P < 0.05) higher than that of the other two groups, but total carnitine excretion was not different among the groups. This study demonstrated that Korean immature and preterm newborns have higher serum triacylglycerol concentrations but lower carnitine status than NBWI. Therefore, the lower carnitine status and moderately higher triacylglycerols may suggest that LBWI in Korea might be at risk for poor carnitine status and decreased capacity to utilise fatty acids for energy. PMID- 17403273 TI - Influence of cabbage processing methods and prebiotic manipulation of colonic microflora on glucosinolate breakdown in man. AB - Glucosinolate consumption from brassica vegetables has been implicated in reduction of cancer risk. The isothiocyanate breakdown products of glucosinolates appear to be particularly important as chemoprotective agents. Before consumption, brassica vegetables are generally cooked, causing the plant enzyme, myrosinase, to be denatured, influencing the profile of glucosinolate breakdown products produced. Some human intestinal microflora species show myrosinase-like activity (e.g. bifidobacteria). We aimed to increase bifidobacteria by offering a prebiotic (inulin) in a randomised crossover study. Six volunteers consumed inulin (10 g/d) for 21 d followed by a 21 d control period (no inulin). Treatment periods were reversed for the remaining six volunteers. During the last 5 d of each period two cabbage-containing meals were consumed. Total urine output was collected for 24 h following each meal. Cabbage was microwaved for 2 min (lightly cooked) or 5.5 min (fully cooked). Faecal samples were collected at the start and after the inulin and control treatments. Bifidobacteria were enumerated by real time PCR. Allyl isothiocyanate production was quantified by measuring urinary excretion of allyl mercapturic acid (AMA). Bifidobacteria increased following prebiotic supplementation (P < 0.001) but there was no impact of this increase on AMA excretion. AMA excretion was greater following consumption of lightly cooked cabbage irrespective of prebiotic treatment (P < 0.001). In conclusion, the most effective way to increase isothiocyanate production may be to limit the length of time that brassica vegetables are cooked prior to consumption. PMID- 17403274 TI - Exogenous nucleic acids and nucleotides are efficiently hydrolysed and taken up as nucleosides by intestinal explants from suckling piglets. AB - Human milk is a rich source of RNA, free nucleotides (NT) and nucleosides (NS). To determine the uptake of different NS sources by the intestinal epithelium, jejunal explants from suckling piglets were cultured in a medium supplemented with a mixture of NS (adenosine, cytidine, guanosine, inosine, uridine; 10 mg/l each), a mixture of five NT (AMP, CMP, GMP, IMP, UMP; 7 mg/l each) or RNA (60 mg/l), respectively. Aliquots from the media were taken at different times (0.5, 2, 5, 15, 30, 60, 180 min). NS and NT concentrations were analysed in the different supernatants at those periods using solid-phase extraction followed by HPLC. When explants were cultured in the presence of NS the concentration of these compounds, excepting cytidine, rapidly decreased, suggesting that they are efficiently taken up. When explants were incubated in the presence of NT, the total concentration of these compounds decreased while the total concentration of NS increased, suggesting that enterocytes efficiently hydrolyse NT into NS. Likewise, when explants were incubated in the presence of RNA, the total concentration of both NT and NS increased, indicating that intestinal explants are able to hydrolyse RNA to NT and then to NS in the absence of luminal enzymes. In conclusion, the jejunum of piglets at weaning is able to hydrolyse RNA and free NT to NS, and NS, excepting cytidine, are efficiently taken up by the small intestine. These results suggest that the current concentration of NT used to supplement infant formulas should be reconsidered. PMID- 17403275 TI - Brushing cytology in cutaneous lesions of the head and neck. AB - BACKGROUND: Brushing cytology is a well established diagnostic procedure used by gynaecologists, physicians and surgeons to obtain representative samples from lesions. Our aim was to evaluate its reliability in ulcerative and tumour-like conditions arising in the skin of the head and neck. METHODS: Over 28 months, 86 patients with suspected cutaneous malignant lesions underwent a cytological examination with a cytobrush within the otolaryngology department. RESULTS: Cytological analysis identified 63 out of 64 histologically documented malignant tumours (60 primary basal cell and squamous cell carcinomas and three metastatic adenocarcinomas), and 21 out of 22 benign lesions. There was one false positive and one false negative result. CONCLUSIONS: Brushing cytology of suspected cutaneous malignant lesions is a rapid and reliable diagnostic method which helps the clinician to decide on appropriate planning and treatment. The technique can be performed as an out-patient procedure, and smear preparation can be done in the laboratory, even at a peripheral hospital. PMID- 17403276 TI - Obstructive sleep apnoea adenotonsillectomy in children: when to refer to a centre with a paediatric intensive care unit? AB - OBJECTIVE: To identify regional surgical referral patterns for adenotonsillectomy in children with obstructive sleep apnoea to our tertiary centre with paediatric intensive care unit facilities and to establish guidelines for elective paediatric intensive care unit referral and admission. METHODS: Two methods were used. A questionnaire was sent to ENT consultants in five surrounding hospitals with no in-house paediatric intensive care facilities. The second was a prospective observational study undertaken in our tertiary centre for a sub-set of patients undergoing obstructive sleep apnoea adenotonsillectomy between January 2002 and February 2005. These children were considered high risk as judged clinically by an ENT surgeon. Most had obstructive sleep apnoea and a co morbidity. Otherwise healthy children with simple obstructive sleep apnoea were excluded. RESULTS: 15 out of 20 consultants responded to the questionnaire. Four referred on the grounds of clinical history, five referred based on pulse oximetry, nine referred syndromal children and four did not refer electively. Of the 49 high risk patients operated on, only 12 required paediatric intensive care admission with no emergency paediatric intensive care admissions. No otherwise healthy children with uncomplicated obstructive sleep apnoea symptoms required paediatric intensive care admission during the study period. CONCLUSION: There was no regional consensus regarding paediatric intensive care unit referral for obstructive sleep apnoea adenotonsillectomy. Clinical judgement without complex sleep studies by those experienced in this area was sufficient to detect complicated cases of obstructive sleep apnoea with co-morbidity requiring paediatric intensive care. PMID- 17403277 TI - Oropharyngeal and hypopharyngeal myxoma: case report and literature review. AB - OBJECTIVES: To describe the features of an oropharyngeal and hypopharyngeal myxoma. MATERIALS AND METHODS: Case report of a 34-year-old patient operated upon for a dual-location tumour, and review of the literature. RESULTS: The myxoma is a rare tumour. Various head and neck locations have been described, but not (to our knowledge) a tumour in both the oropharynx and the hypopharynx. Multiple synchronous locations must be searched for, particularly regarding cardiac myxoma. CONCLUSION: The myxoma is a rare, benign tumour, even rarer in the head and neck. Surgical treatment must be complete in order to avoid recurrences, and should be performed after assessment for cardiac involvement. PMID- 17403278 TI - [To develop a multi-center cooperative research system for pancreatic cancer in China]. PMID- 17403279 TI - [Retroactive evaluation of perioperative staging and surgical decision making for patients with pancreatic cancer]. PMID- 17403280 TI - [Therapeutic principles in treatment of severe acute pancreatitis]. PMID- 17403281 TI - [Standard with extended pancreaticoduodenectomy for adenocarcinoma of the head of the pancreas: a meta-analysis]. AB - OBJECTIVE: To compare standard with extended pancreaticoduodenectomy for adenocarcinoma of the head of the pancreas: a meta-analysis of randomized controlled trials and prospective studies. METHODS: Randomized controlled trials and prospective studies comparing standard with extended pancreaticoduodenectomy for pancreatic cancer of head were identified using a systematic search of Medline, the Cochrane Library Databases and CBMDisc covering articles published from 1996 to 2005. Recommendations were based on the available level of evidence (A, large randomized; B, small randomized; C, prospective trial). A fixed-effect model and a random-effect model used vary with the heterogeneity test. Outcome of primary interest was operative morbidity, mortality and survival rates as well. RESULTS: Six RCTs trials and five prospective studies were included. Combined odds ratio for overall morbidity using random effect model was 1.82 (95% CI = 0.68 to 4.90) and OR of overall mortality, 1, 3, 5-year survival rate using fixed effect model was 0.84 (95% CI = 0.28 to 2.55), 0.74 (95% CI = 0.45 to 1.22), 0.90 (95% CI = 0.54 to 1.50), 0.90 (95% CI = 0.54 to 1.50), 1.43 (95% CI = 0.45 to 4.55) respectively. and indicated no significant difference. CONCLUSIONS: No evidence was found that extended pancreaticoduodenectomy leads to longer survival than standard group (A level). There is no significant difference between standard and extended group in morbidity and mortality. Whipple procedure is also of choice for pancreatic head carcinoma and extended pancreaticoduodenectomy is indicated for lymph node positive patient (A-level). PMID- 17403282 TI - [Study on the quality of life after pancreaticoduodenectomy]. AB - OBJECTIVE: To evaluate the quality of life (QOL) of the patients who received pancreaticoduodenectomy (PD) and work out their long term therapy agents. METHODS: QOL of 18 cases who received PD (group PD) and 18 cases received laparoscopic cholecystectomy (LC) (group LC) in the same days was determined by symptoms questionnaire and Chinese version SF-36 QOL questionnaire from Jan 2002 to Dec 2003 in Sir Run Run Shaw Hospital. RESULTS: Compared with group LC, the total QOL score and physical health score of group PD didn't have significant decrease. But the mental health score of group PD was lower than group LC (P < 0.05). Eight different scales of SF-36 questionnaire showed that the score in physical functioning, role-physical, bodily pain, general health, social functioning of group PD was lower than that of group LC. The score in vitality, role-emotional and mental health of group PD was the same as the group LC. According to the symptoms questionnaire, the patient diarrhea and recurrence had obvious influence on PH score. The patient weight loss and unemployment had obvious influence on MH2 score. CONCLUSIONS: The QQL of patients received PD didn't have decreased. Their total score of SF-36 QQL was close to the patients who received LC. But the mental health score of group PD was lower than group LC. Weight loss, unemployment, recurrence and chronic pancreatic diarrhea may be infect the Quality of life after PD. PMID- 17403283 TI - [Clinical evaluation of 21 cases of total pancreatectomy]. AB - OBJECTIVE: To evaluate the clinical outcome of 21 cases of total pancreatectomy. METHODS: The clinical data of 21 cases of total pancreatectomy performed from April 2003 to June 2006 was retrospectively analyzed. RESULTS: Among the 21 patients, 1 case combined with transverse colon resection, 1 case with total gastrectomy, 9 cases with portal-superior mesentery vein resection with end-to end anastomosis, 9 cases with portal-superior mesentery vein resection and grafts implantation, 8 cases with concomitant celiac axis resection, 4 cases with concomitant celiac axis and common hepatic artery resection, 1 case with concomitant celiac axis, portal vein and superior mesentery artery resection and reconstruction. Complications occurred in 12 cases (57.1%) post the operation and 5 cases (23.8%) died in 30 days after the operation. Insulin was given at the dose of 18 - 28 U daily post operation and blood glucose was maintained normal effectively. Sixteen cases were followed-up and median survival was 9.2 months (1.2 - 13.0 months). The median survival of tubular adenocarcinoma and intraductal papillary mucinous neoplasms of the pancreas (IPMNs) were 7 months (1.2 - 9.0 months) and 11.3 months (10.0 - 13.0 months), respectively. CONCLUSIONS: Total pancreatectomy could not improve survival and it increases the complications and death, but it could improve the quality of life. It's an operation of choice for IPMNs, but with pancreatic carcinoma, the warranty of operation should be considered. The blood glucose level could be maintained normal effectively after the operation. PMID- 17403284 TI - [Indication and choice of operation technique for duodenum-preserving resection of pancreatic head: 22 cases reports]. AB - OBJECTIVE: To analyze the indication and choice of operation technique for duodenum-preserving resection of pancreatic head. METHODS: The Clinical material of the 22 patients who received duodenum-preserving resection of pancreatic head (DPPHR) from January 2001 to January 2006 was analyzed. Of the 22 cases, 8 cases presented with mucinous cystadenoma, 2 cases with mucinous cystadenocarcinoma, 4 cases with solid-pseudopapillary tumors, 2 cases with pancreatic endocrine tumors, 4 cases with chronic pancreatitis, 1 case with lymph epidermis cyst, 1 case with serous cystadenoma. The indication, choice of operation technique of DPPHR and the prevention and management of the post-operative complications were investigated. RESULTS: No patient died of the operation. Three cases (13.6%) developed pancreatic fistula after the operation, 1 case (4.5%) developed biliary fistula, 1 case (4.5%) developed abdominal infection and 2 cases of duodenal fistula occurred (9.1%). CONCLUSIONS: DPPHR retains the continuity of stomach, duodenum and biliary ducts. The operation is safe and it reduces wounds and excision scope. This procedure can be used in benign and low malignant lesions in the head and neck of the pancreas. PMID- 17403285 TI - [Treatment of late-staged pancreatic carcinoma with implantation of 125I seeds in combination with regional arterial infusion chemotherapy]. AB - OBJECTIVE: To evaluate the effect of combined use of implantation of (125)I seeds with regional arterial infusion chemotherapy on late-staged pancreatic carcinoma. METHODS: During operation, (125)I seeds were implanted into tumor parenchyma or operative beds for 7 patients with pathological diagnosis of late-staged pancreatic carcinoma, then chemotherapy pump was implanted into common hepatic artery or celiac artery through left gastric artery or right epigastric artery. Gemcitabine (1000 mg/m(2)) plus fluorouracil (500 mg/m(2)) was used for postoperative adjuvant chemotherapy once a week for 7 consecutive weeks. RESULTS: The mean number of implanted (125)I seeds was 42 and the mean dose was 29.4 mCi. The mean follow-up time was 245 days. Rate of pain relief was 100%, metastasis foci was reduced in 2 cases. CA-19-9 Antigen level decreased to normal in all patients. Pancreatic fistula occurred in one case and was cured with conservative therapy. CONCLUSIONS: The combination therapy is effective for late-staged pancreatic carcinoma, it brings symptom relief and improvement in quality of life. PMID- 17403286 TI - [Surgical effect of malignant tumor of body and tail of the pancreas: compare with pancreatic head cancer]. AB - OBJECTIVES: To investigate the clinical-pathological characteristics and surgical prognosis of malignant tumor of pancreatic body and tail. METHODS: A retrospective study was accomplished on clinical manifestation, pathological behavior and postoperative survival in 106 patients with malignant tumor of pancreatic body and tail in single institution from Jan 1980 to Dec 2003, and compared these with 451 patients with malignant pancreatic cancer. RESULTS: There were significant differences in the following parameters (malignant tumor of the body and tail vs those of the head) between the two tumors: (1) the complaints of pain (0.74:41, chi(2) = 37.035, P < 0.01) and jaundice (0.04:0.75, chi(2) = 155.509, P < 0.01); (2) serum SGPT [(27.33 +/- 3.98) U/L: (118.60 +/- 4.59) U/L, F = 89.351, P < 0.01], total bilirubin [(1.46 +/- 0.46) mg/dl: (14.11 +/- 0.60) mg/dl, F = 105.341, P < 0.01] and albumin [(4.20 +/- 0.45) g/L: (3.91 +/- 0.03) g/L, F = 26.642, P < 0.001]; (3) CEA (0.40:0.24, chi(2) = 6.148, P = 0.046) and CA-19-9 positive rate (0.57:0.86, chi(2) = 24.132, P < 0.01); (4) the concomitant total metastasis (0.38:0.20, chi(2) = 14.266, P < 0.01), including liver metastasis (0.30:0.17, chi(2) = 9.003, P < 0.01). Postoperative median survival, resection of non-metastatic pancreatic body and tail cancer was longer than resection of metastatic disease significantly (15 vs 7 months,chi(2) = 21.63, P < 0.01), which the latter was the same as those who didn't remove (6 months,chi(2) = 0.22, P = 0.64). CONCLUSIONS: The predominant problem is distant metastasis (especially liver metastasis) in the malignant tumor of the body and tail of the pancreas in comparison with pancreatic head cancer. Resection of the body and tail could not increase postoperative survival if metastasis exists. The major way to improve the prognosis is to prevent and manage the distant metastasis. PMID- 17403287 TI - [Immuno-proteomic screening of human pancreatic cancer associated membrane antigens for early diagnosis]. AB - OBJECTIVE: To screen and identify the immunogenic membrane antigens in human pancreatic cancer for early diagnosis. METHODS: Membrane protein was extracted from pancreatic cancer cell lines and separated by using 2-DE. One of the two parallel 2-DE gels went for staining while the other underwent immunoblot. Serum IgG, which was purified from clinically collected sera of pancreatic cancer patients, was used as the primary antibodies for the immunoblot. Positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and PMF matching, and then evaluated by bio-informatics methods. The candidate membrane antigens were further validated respectively in cell lines and tissues by RT-PCR and immunohistochemistry. RESULTS: The immunoblot of mixed membrane protein with serum IgG from cancer patients showed eight positive dots. These dots were identified with MALDI and PMF as: VDAC-1, VDAC-2, CHCHD3, SLP-2 and TOM40. RT-PCR showed that these membrane antigens were expressed in several pancreatic cancer cell lines. Immunohistochemistry showed prominent SLP-2 over expression in cancer tissue. CONCLUSIONS: VDAC-1, VDAC-2, CHCHD3, SLP-2, and TOM40 are the new candidate immunogenic membrane antigens of pancreatic cancer. These membrane antigens can be subsequently tested in high dangerous population for early diagnosis of pancreatic cancer. PMID- 17403288 TI - [Construction and primary application of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes]. AB - OBJECTIVE: To investigate the construction of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes and its application. METHODS: Pancreatic cancer related genes were purposely selected, and oligonucleotide microarray was prepared by spotting oligonucleotide probes onto glass slides coated with APS-PDC. Total RNA were extracted from frozen tissues with TRIzol method according to the manufacturer's protocol, and purified with QIAGEN RNeasy Kit. Labeled cDNA targets for hybridizations were synthesized by reverse transcription from control- and cancer-total RNA samples in the presence of Cy5-dCTP and Cy3-dCTP, respectively. The labeled probes were hybridized with oligonucleotide microarray for 16 h to 18 h. Hybridized microarray was scanned by Agilent laser scanner, and the acquired image was analyzed by Imagene3.0 software. The intensity ratio of Cy3 and Cy5 were calculated. To confirm the expression profiles of these genes, quantitative reverse transcription-PCR (Q RT PCR) was carried out with CDC25B and TUSC3 genes. The product of PCR were quantitated by comparative Ct method. RESULTS: The signal of microarray hybridization was clear, and the images had a lower background and higher signal noise ratio. The signal of positive control spots were uniform, and spots of negative control and blank signal were fairly low. In comparison with normal pancreas, 24 differential expressed genes were identified, which included 17 up regulated and 7 down-regulated genes. The results of Q RT-PCR demonstrated that the expression of CDC25B and TUSC3 in pancreatic cancer were increased and decreased respectively, which consistent with microarray hybridization. CONCLUSIONS: The oligonucleotide microarray specialized for pancreatic cancer are desirable for its specialty, flexibility and sensitivity, which can simultaneously and parallelly detect multiple pancreatic cancer-associated genes. In contrast to normal pancreatic tissues, the genes expression profile are different significantly in pancreatic cancer. PMID- 17403292 TI - [Significances of gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time]. AB - OBJECTIVE: To investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time. METHODS: The liver tissues were divided into two groups: Group A (non hepatocirrhosis), Group B (hepatocirrhosis), each of which consisted of 3 groups with different ischemic time: 15, 30 and 45 minutes. The gene differential expression patterns in the two groups within different ischemic time were detected and compared with those in normal liver tissues by using 4000 points gene microarray. RESULTS: In non-hepatocirrhosis tissues, the homeostatic maintenance genes expressed highly during hepatic ischemia for 15 minutes, and no apoptotic gene was expressed; but in hepatocirrhosis tissues, many apoptotic genes expressed highly. As for 30 minutes, in both two groups liver tissue genes expressed to the peak, and the genes related to cell death, oxidative stress and nuclear factors expressed highly. The difference lies in the facts that in Group B pro-apoptosis genes expressed more than those in Group A, and the Ratio values were higher than those in Group A. Many genes of heat shock protein family and antioxidant proteins expressed highly simultaneously in Group A, but comparatively low in Group B. As for 45 minutes, genes of heat shock proteins and antioxidant proteins expressed lowly in Group B. CONCLUSIONS: It suggests that the safe time limit of hepatic ischemia for cell survive is 30 minutes or so. Non hepatocirrhosis tissues could endure 30 minutes of ischemia and even longer, but it should be restricted within 30 minutes in hepatocirrhosis tissues. PMID- 17403293 TI - [Evaluation of cyclooxygenase-2 in the prediction of liver metastasis of colorectal carcinoma]. AB - OBJECTIVE: To investigate the expression of cyclooxygenase-2 (COX-2) in colorectal carcinoma and its correlation with liver metastasis of colorectal carcinoma. METHODS: The expression of COX-2 was detected using immunohistochemical methods in 30 colorectal carcinoma tissues without liver metastasis, 30 with preoperative liver metastasis, 30 with postoperative liver metastasis and 30 surrounding normal colorectal tissues. And its correlation with gender, age, Dukes stages was analyzed too. RESULTS: The expression of COX-2 in colorectal carcinoma was significantly higher than that in surrounding normal colorectal tissue (P < 0.05), and meanwhile, its level in colorectal carcinoma without liver metastasis was significantly lower than those in tissues with preoperative or postoperative liver metastasis (P < 0.05). The COX-2 level had no correlation with gender, age, histological type, histological grade or the preoperative serum CEA and CA 19-9 levels in colorectal carcinoma (P > 0.05), but it was related to Dukes stages and lymph node metastasis. CONCLUSIONS: COX-2 plays a role in the course of generation, development and metastasis of colorectal carcinoma. The high expression of COX-2 in colorectal carcinoma tissues may be considered as an indicator for liver metastasis. PMID- 17403294 TI - [The relationship between Oddi's sphincter and bile duct pigment gallstone]. AB - OBJECTIVE: To investigate the relationship between anatomic abnormalities and malfunction of Oddi sphincter with formation of bile duct pigment gallstone. METHODS: One hundred and twenty-three patients with a T tube after cholecystectomy and choledochotomy were divided into reflux group and control group by measuring the amounts of radioactivity of (99m)Tc-DTPA in the bile. Among them 53 were selected randomly to undergo choledochoscopic manometry. Basal pressure of Oddi's sphincter (SOBP), amplitude of Oddi's sphincter (SOCA), frequency of contraction (SOF), duration of contraction (SOD), duodenal pressure (DP), common bile duct pressure (CBDP) were scored and analyzed. The level of plasma motilin and serum gastrin of 45 patients and 12 healthy volunteers were measured by radioimmunoassay. The incidence rates of duodenal descending part diverticulum in patients with bile duct pigment stones, patients without alimentary tract diseases, patients with gallbladder polyps, patients with gallbladder stones were studied by means of barium meal examination. The incidence rates of intraduodenal peri-ampullary diverticulum in patients with primary bile duct pigment stones, patients with bile duct stone and gallbladder stones, patients with bile duct stones originating from the gallbladder, patients with inflammation and stricture of the extremity of bile duct and papilla, patients with cancer of the extremity of bile duct and papilla, patients with post-cholecystectomy syndrome were detected by duodenoscope. RESULTS: Of the patients, 44 were detected with duodenal-biliary reflux (35.8%). SOBP, SOCA and CBDP in the reflux group were much lower than those in control group (P < 0.001). The level of serum gastrin and plasma motilin of the reflux group were much lower than those of control group (P < 0.01). Positive correlation was found between level of motilin and SOBP while level of gastrin was positively correlated with SOBP and CBDP. The incidence of duodenal diverticulum in patients with bile duct pigment stone was 36.62%, which was higher than that of the other 3 groups. The incidence rate of intraduodenal peri-ampullary diverticulum in patients with primary bile duct pigment stone was higher than that of patients with inflammation and stricture of the extremity of bile duct and papilla, patients with cancer of the extremity of bile duct and papilla and patients with bile duct stones originating from the gallbladder. CONCLUSIONS: The patients with bile duct pigment stone have apparent duodenal-biliary reflux and infection of the bile duct. The state of structure and function of Oddi's sphincter is correlated significantly with bile duct pigment stone. The anatomic abnormalities and malfunction of Oddi's sphincter played an important role in the formation of bile duct pigment stone. PMID- 17403295 TI - [Role of c-Jun NH (2)-terminal kinase in insulin resistance after burn]. AB - OBJECTIVE: To investigate the role of c-Jun NH (2)-terminal kinase (JNk) in insulin resistance after burn and its mechanism. METHODS: Twenty-four Sprague Dawley rats were randomized to control, burn and burn + anisomycin groups. The rats in control group received sham burn trauma, and burn and burn + anisomycin groups received 30% total body surface area (TBSA) full thickness burn injury. Anisomycin (5 mg/kg) together with 250 microl dimethyl sulfoxide (DMSO) was injected to the rats in anisomycin group intravenously, and only 250 microl DMSO in the other two groups. Euglycemic-hyperinsulinemic glucose clamps was performed 2 hours after the injection. The changes of phospho-serine 307, phospho-tyrosine of insulin receptor substrate (IRS)-1 and phospho-JNK in muscle tissues were determined and compared using immunoprecipitation and Western blot analysis or immunohistochemistry in the three groups. RESULTS: The infusing rates of total 10% glucose (mg x kg(-1) x min(-1)) in control, burn and burn + anisomycin group were 12.3 +/- 0.4, 6.6 +/- 0.3, 6.5 +/- 0.4, respectively. The level of IRS-1 Serine 307 phosphorylation and phospho-JNK in muscle increased significantly, while insulin-induced tyrosine phosphorylation of IRS-1 decreased markedly after burn. CONCLUSIONS: The activation of JNK elevates the level of IRS-1 phospho serine 307 and might play a role in insulin resistance after burn in rats. PMID- 17403301 TI - [Ethical principles in the publication of biomedical research articles]. PMID- 17403302 TI - [Important advances and restricted factors in anti-virus therapy in patients with chronic hepatitis B]. PMID- 17403303 TI - [Correlation of anti-60000 SSA antibody and anti-52000 SSA antibody with systemic lupus erythematosus and Sjogren's syndrome]. AB - OBJECTIVE: To investigate the correlation of anti-60 000 SSA antibody and anti-52 000 SSA antibody with systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS). METHODS: Western blotting against purified 60 000 SSA antigenic polypeptide and 52 000 SSA antigenic polypeptide were done to detect the anti-60 000 SSA and anti-52 000 SSA antibodies in 59 serum samples positive in anti-SSA antibodies, among which 44 samples were from SLE patients and 15 samples from SS patients. RESULT: There was no significant difference in positive rate of anti-60 000 SSA antibodies between the SLE and pSS patients (P > 0.05). But the sole positive rate of anti-60 000 SSA antibodies of the SLA patients was 39.47% (15/38), significantly higher than that of the primary SS (pSS) patients ((6.67%, 1/15, P < 0.05). The positive rate of anti-52 000 SSA antibodies of the pSS patients was 93.33% (14/15), significantly higher than that of the SLE patients (23/38, 60.53%, P < 0.05). There was no significant differences in positive rates of anti 60 000 SSA antibodies and anti-52 000 SSA antibodies between the secondary SS (sSS) and pSS patients (P > 0.05). CONCLUSION: The pSS patients mainly present anti-52 000 antibodies and with a very low sole anti-60 000 SSA antibody positive rate. Sole positivity of anti-52 000 SSA antibodies maybe correlated with pSS and is not a strong implication for the diagnosis of sSS. Sole positive anti-60000 SSA antibodies rate can be seen mainly in SLE. PMID- 17403304 TI - [The significance and characteristics of chromosomal abnormalities in patients with microsatellite and chromosome stable colorectal carcinoma]. AB - OBJECTIVE: To investigate the clinico-pathological significance and characteristics of chromosomal abnormalities in microsatellite and chromosome stable (AMCS) colorectal carcinoma (CRC). METHODS: Flow cytometry, microsatellite instability analysis and immunohistochemistry methods were used to examine the DNA ploidy, microsatellite instability and expression of mismatch repair proteins (hMSH(2) and hMSH(1)) in 156 cases of CRCs, so as to select the MACS CRCs. Comparative genomic hybridization (CGH) method was subsequently performed to analyze the status of chromosomal abnormalities of MACS CRCs. The correlation between chromosomal changes of MACS CRC and patients clinico-pathological features was further evaluated. RESULTS: Of the 156 CRCs studied, fourty-one (26%) cases were observed both diploid/near diploid DNA content and stable microsatellite sequence and all these CRCs showed normal expression of hMSH(2) and hMSH(1) protein, and thus, defined as MACS CRC. Our CGH results showed that the overall mean numbers of chromosomal abnormality of 41 MACS CRCs was 9.6. The chromosomal arms with DNA amplification (frequency > or = 10%) were 20 q (68%), 13 q (56%), 7 q (49%), 13 p (39%), 20 p (37%), 8 q (30%), 1 q (22%), 11 q (15%), 16 q (12%) and 2 p, 4 q, 10 q (10%). The chromosomal arms with DNA deletion (frequency > or = 10%) were 18 q (63%), 8 p (51%), 17 p (37%), 1 p (30%), 3 p (26%), 4 p, 13 q, 14 (15%) and 21 q, Xp (10%). In addition, there was 79% and 82% MACS CRCs in Dukes'C/D stages observed amplification of 20 q and deletion of 18 q, respectively, the frequency was significantly higher that that in Dukes'A/B stages (46%, P = 0.038 and 21%, P = 0.0009). CONCLUSIONS: The overall number of chromosomal abnormalities in MACS CRCs is similar to that of chromosomal instable (CI) CRCs, but is higher than that of microsatellite instable (MSI) CRCs. The frequent deletion of 8 p in MACS CRCs might be a special molecular event that is different from CRCs in CI and MSI pathway. Both 20 q amplification and 18 q deletion are the most frequent chromosomal abnormalities in MACS CRC and are associated closely with tumor's malignant clinical phenotype. These chromosomal changes might play important roles the development and progression of MACS CRC. PMID- 17403305 TI - [Effect of tumor burden on differentiation of T lymphocytes in the peripheral blood of patients with colorectal cancer]. AB - OBJECTIVE: To analyze the effect of tumor burden on the differentiation of T1 and T2 cells and its implication in patients with colorectal cancer. METHODS: Peripheral venous blood samples were obtained from 20 patients with primary colorectal cancer before and 7 days after the operation, radical operation in 17 patients and palliative resection in 3 patients. Twenty sex and age-matched patients with benign diseases treated in the same period were used as controls. T1 and T2 cells in the peripheral blood were evaluated by detecting the intracellular interferon-gamma and interleukin-4 production with 4-color flow cytometry. Lymphocyte subsets in the peripheral blood were also measured by flow cytometry. RESULTS: At the time of diagnosis, the percentage of T1 and T2 cells in the peripheral lymphocytes of the case group was lower significantly than that of the control group (P = 0.006, and P = 0.017). There was no significant difference in the T, CD4(+) T, CD8(+) T, B, and NK cells between the two groups. After getting rid of the tumor burden, the percentage of T1 cells increased, however, not significantly (P > 0.05). And the percentage of T2 cells increased significantly (P = 0.020). The percentages of T1 cell of the patients with the tumor > or = 5 cm and of the patients with poorly differentiated tumors were significantly lower than those of the patients with the tumor < 5 cm and of the patients with well or moderately differentiated tumors (P = 0.064, and P = 0.072). The percentage of T1 cells in the patients with lymph node metastasis and stage III approximately IV tumor were lower significantly than those of the patients without lymph node metastasis and those with stage I approximately II tumor (P = 0.033 and P = 0.033). No significant differences were found between the population of T1 cells and such factors as tumor size, serosa invasion, and distant metastasis. CONCLUSION: Tumor load suppresses the differentiation of T1 and T2 cells in the patients with colorectal cancer significantly, and may be an important factor in the development of immunosuppression. After getting rid of the tumor burden, the percentages of T1 and T2 increase in a short time, and the immunologic function is improved. Determination of T1 may be helpful to indicate the prognosis of colorectal cancer. PMID- 17403306 TI - [Influence of different operation modes in treatment of leiomyoma on reproductive endocrine hormone levels]. AB - OBJECTIVE: To explore the most reasonable operation mode for treatment of leiomyoma so as to protect the ovary function of the patients. METHODS: 103 leiomyoma patients under the age of 49 received different operation: myomectomy (Group I, n = 33), subtotal hysterectomy (Group II, n = 30), and hysterectomy (Group III, n = 40). The levels of serum estradiol (E(2)), progesterone (P), luteinizing hormone (LH), and follicle stimulating hormone (FSH) were detected before operations and 3 and 6 months after operations. Sixty patients underwent color Doppler ultrasonography before and after hysterectomy or subtotal hysterectomy to measure the blood flow of ovary artery. The maximal systolic flow velocity (Vmax), end-diastolic minimal flow velocity (Vmin), resistance index (RI), and pulsatility index (PI) were calculated. RESULTS: There were no significant differences in the levels of E(2), P, LH, and FSH before and after operation among the patients of Group I regardless of the age (all P > 0.05) The E(2) levels 6 months after operation of Group II and Group III were both significantly lower than those before operation (both P < 0.05), however, the P, LH, and FSH levels before and after operation were not significantly different in these 2 groups (all P > 0.05). The E(2) level 6 months after operation of the patients aged > or = 40 in group III decreased much more significantly to (362 +/ 252) pmol/L, with a greater statistical difference in comparison with that before operation, (567 +/- 417) pmol/L (P < 0.01). The values of Vmax and Vmin of the ovary artery 6 months after operation were: 0.24 +/- 0.04 m/s and 0.05 +/- 0.05 m/s respectively, both significantly lower than those before operation, (0.50 +/- 0.11 m/s and 0.17 +/- 0.24 m/s respectively, both P < 0.01). The values of RI and PI 6 months after operation were: 0.80 +/- 0.05 and 2.06 +/- 0.24 respectively, both significantly higher than those before operation (0.74 +/- 0.05 and 1.62 +/- 0.33 respectively, both P < 0.01). CONCLUSION: Myomectomy doesn't influence the ovary function. Both subtotal hysterectomy and hysterectomy decrease the ovary blood flow and endocrine function 6 months after operations. PMID- 17403307 TI - [Analysis of peripheral arterial obstructive disease related factors among diabetic population aged > or = 50]. AB - OBJECTIVE: To analyze the peripheral arterial obstructive disease (PAD) related factors among diabetic population aged > or = 50 in China. METHODS: The clinical data of 1397 diabetic patients aged > or = 50 with at least one of the following risk factors: smoking, hypertension, and hyperlipidemia, from 15 Class III Grade A hospitals in 7 major cities of China were collected. Diagnosis of PAD was based on the ankle brachial index (ABI) < 0.9, and diagnosis of arteriosclerosis was based on pulse wave velocity (PWV) > 1400 cm/s. Regression studies were made to analyze the relations among PAD and various risk factors: age, sex, body mass index (BMI), smoking, hypertension, hyperlipidemia, history of cerebral vascular disease (CVD), history of ischemia heart disease (IHD) etc. RESULTS: The current prevalence rate of PAD was 19.47% among the 1397 patients, 18.3% (122/664) among the male patients, and 20.4% (150/733) among the female patients. The prevalence of PAD in the patients aged > or = 70 was as high as 31.9%. The duration of diabetes course was positively correlated with the prevalence of PAD (chi2 = 11.9, P = 0.0026). The ABI abnormality rate was 15.78% among those with a diabetic course of 5 years and was 23.84% among those with a diabetic course of 10 years. The abnormal ABI rate of the patients with CVD was 30.57%, significantly higher than that of hose without CVD (17.29%, chi2 = 21.49, P < 0.0001). The abnormal ABI rate of the patients with IHD was 24.64%, significantly higher than that of the patients without IHD (18.20%, chi2 = 5.85, P = 0.0155). The HbA1c value of the PAD patients was significantly higher than that of the patients without PAD (chi2 = 5.10, P = 0.0239) Odd risk analysis showed that age increase of 10 years increased the PAD risk by 1.64 times (OR = 1.6444, P = 0.0001). The PAD risk of the smokers was 1.68 times higher than that of the non smokers (OR = 1.6852, P = 0.0001). Increase of 10 mm Hg in systolic blood pressure (SBP) increased the PAD risk by 1.19 times (OR = 1.1926, P = 0.01). The PAD risk of the patients with abnormal HbAlc was 2.44 times higher than that of the patients with normal HbA1c (OR = 2.4473, P = 0.0001). One-year's increase of the hypertension course increased the PAD risk by 1.02 times (OR = 1.0194, P = 0.03). Logistic analysis indicated that the relations among PWV and the risk factors were almost the same among ABI abnormality and the risk factors. CONCLUSION: Approximately one fifth of diabetic patients aged > or = 50 in China suffer from PAD. Age, course of diabetes, blood glucose level, SBP, IHD, and CVD are risk factors for PAD. Early intervention and treatment of hypertension and hyperglycemia, and quitting smoking are important in reducing the occurrence of PAD. ABI and PWV are not only diagnostic means for PAD, but also alarm guide indexes for cerebral vascular disease (CVD). PMID- 17403308 TI - [Treatment of cervical spondylotic myelopathy by decompression of spinal canal and internal fixation with the combination of anterior and posterior approaches]. AB - OBJECTIVES: To evaluate the clinical results of treatment of cervical spondylotic myelopathy by decompression of spinal canal and internal fixation with the combination of anterior and posterior approaches, and to study the methodology to prevent operative complications. METHODS: 28 consecutive cases with cervical spondylotic myelopathy, aged 65.3 (49 ~ 73) were treated by decompression of spinal canal and internal fixation with the combination of anterior and posterior approaches from December 2001 to December 2005. The surgical methods included decompression of spinal canal, spinal fusion, and internal fixation with the combination of anterior and posterior approaches. The patients were followed up for 16 months (6 ~ 52 months). The clinical results and complications were analyzed retrospectively. The clinical effects were evaluated by the Japanese Orthopedic Association (JOA) scoring system and Odom's criteria. The outcome of cervical spinal fusion was evaluated by X-ray plate. RESULTS: There were no neural injury and wound healing problems in all patients. All postoperative patients ambulated three days after the operation. CSF leak occurring in 3 cases was cured by conservative treatment. The recovery rate by the JOA scoring system was 50% when the patients were discharged. According to the Odom scoring system, 83.3% of the patients had excellent and good effects. No evidence of implant failure was found according to the Brantigan's criteria. Interbody fusion was achieved in 3 approximately 6 months after operation in all followed-up patients. No additional operation was needed for the involved segments, the spinal cord function of the patients had been improved in a certain extent, and the surgical outcomes could be maintained. CONCLUSION: Decompression of spinal canal and internal fixation with the combination of anterior and posterior approaches is necessary and effective in the treatment of severe and multi-level cervical spondylotic myelopathy, but the operative trauma is relatively serious, surgeons must carefully design the operative plan and pay enough attention to the methods to prevent operative complications, especially for the elderly patients. PMID- 17403309 TI - [Analysis of the intervertebral angle following anterior spinal fusion of thoracolumbar/lumbar adolescent idiopathic scoliosis]. AB - OBJECTIVE: To investigate the changes of intervertebral angle and relative factors after anterior correction and instrumentation in thoracolumbar/lumbar (PUMCIb, Ic) adolescent idiopathic scoliosis patients. METHODS: The clinical data, including roentgenograms of 18 PUMCIb, Ic adolescent idiopathic scoliosis patients, including 13 patients of Ib type, 2 males and 11 females, and 5 patients of Ic type, all females, 1 who underwent anterior correction and instrumentation, were collected. The Cobb angle of the coronal curve, correction rate, trunk shift, apical vertebra translation, intervertebral angle, and so on, were measured and analyzed. RESULTS: The average coronal Cobb angle were 44.72 +/ 10.20 degrees and 6.51 +/- 7.54 degrees before and after surgery respectively, with an average correction rate of 81.2%. The average preoperative intervertebral angle was 3.57 +/- 3.48 degrees , and 2.02 +/- 4.37 degrees and 5.26 +/- 5.92 degrees 1 week and 1 year postoperatively. The changes were of significance in the intervertebral angle between preoperative and 1-week postoperative values (P = 0.035), and between the 1-week and 1-year postoperative values (P = 0.029). All 18 patients were grouped to Group A (n = 12) and Group B (n = 6) according to the fusion levels. In Group A the preoperative intervertebral angle was 4.07 +/- 3.98 degrees ; the intervertebral angle 1-week and 1-year postoperatively were 1.23 +/ 3.46 degrees and 4.01 +/- 4.72 degrees respectively. In Group B, the preoperative intervertebral angle was 1.97 +/- 1.76 degrees ; and the intervertebral angle 1-week, and 1-year postoperatively were 4.21 +/- 4.47 degrees and 7.12 +/- 5.09 degrees respectively. There were significant changes between the two groups about the intervertebral angle in different stages (P < 0.05). The analysis and comparison of the other variables between these two groups revealed significant changes in the preoperative apical vertebra translocation, number of fused segments and T12 approximately LI lordosis (all P < 0.05). CONCLUSION: The intervertebral angle is much improved after anterior correction and instrumentation surgery, but it increases gradually during the follow-up. The fusion levels, the preoperative apical vertebra translocation, T12 approximately LIV lordosis, and the preoperative intervertebral angle may be correlated with the changes of postoperative intervertebral angle. In the current opinion, it's unworthy of preventing postoperative intervertebral angle changes by decreasing the correction rate or sacrificing the motion segment. PMID- 17403310 TI - [Angiographic outcomes of cerebral aneurysms embolized with Matrix coils]. AB - OBJECTIVE: To investigate the effect of Matrix coil in prevention of recanalization of intracranial aneurysm after embolization with Matrix coils. METHODS: Plan of follow-up was designed to make digital subtraction angiographies for Forty-eight intracranial aneurysm patients with 49 aneurysm s, 22 males and 26 females, aged 52 (15 ~ 67), underwent embolization with Matrix coil and were followed up for 11.75 months on average. Digital subtraction angiography was conducted immediately after the operation and during the follow-up. At least three projections, including anterior-posterior projection, lateral projection, and optimal working projection were adopted. The percentages of occlusion of the initial and the follow-up images were compared. RESULTS: DSA immediately after the operation showed a complete occlusion rate of 34.7%, near complete rate of 42.9% and an incomplete rate of 22.4%. The follow-up DSA revealed that the complete occlusion rate was 53.1%, near complete rate was 24.5%, and incomplete rate was 22.4%. The overall recanalization rate was 28.5%, 22.2% for small aneurysms and 46.1% for large aneurysms. The major recanalization rate was 14.3% and the improvement rate was 24.5%. The recanalization rate was 11.8% for total occlusion, 33.3% for near complete occlusion, and 45.5% for incomplete. CONCLUSION: The long-term angiographic outcome of intracranial aneurysms after embolization with Matrix coils is stable. The recanalization rate is acceptable and is not better than that of GDCs. PMID- 17403311 TI - [Clinicopathological features and prognosis of bronchial mucoepidermoid carcinoma: analysis of 21 cases]. AB - OBJECTIVE: To investigate the clinicopathological features and prognosis of bronchial mucoepidermoid carcinoma. METHODS: Twenty-one with mucoepidermoid carcinoma, 16 males and 5 females, aged 40.8 (15 ~ 72) underwent surgical treatment, 13 cases undergoing lobectomy, 3 sleeve lobectomy, 4 pneumonectomy, and 1 case thoracic exploration, from February 1988 to June 2005. Bronchoscopy was done in all patients before operation. Follow-up was made for 68.3 months on average. RESULTS: Most patients presented the respiratory symptoms such as cough, dyspnea, hemoptysis, and obstructive pneumonia. Biopsy examination through bronchoscopy confirmed the diagnosis of bronchial mucoepidermoid carcinoma in 16 cases. Pathology showed 17 cases of low grade tumor and 4 cases of high grade tumor. Follow-up showed that 17 cases of low-grade group (stage T(1 - 3)N(0 - 1)M(0)) all survived, 8 of which had survived for more than 5 years, including one patient with hilar lymph node metastasis. However, none of the 4 cases with high-grade tumor survived for more than 1 year except one that survived for 13 months. CONCLUSION: Patients with low-grade mucoepidermoid carcinoma can be expected to be cured through curative resection, however, patients in high-grade group have poor prognosis because of metastasis. TNM staging, malignancy grade, and local lymph node metastasis play the key roles in prediction of prognosis. PMID- 17403312 TI - [Impact of propofol on the optimal sedative depth in patients undergoing gastroscopy]. AB - OBJECTIVE: To explore the optimal sedative depth and adequate bispectral index (BIS) value in patients undergoing gastroscopy treated with propofol and to explore the maneuverability and practicability of using BIS value as a variable to control the infusion of propofol. METHODS: 160 patients, 76 males and 84 females, aged 44 +/- 15 voluntarily undergoing painless gastroscopy were randomly divided into 4 equal sex and age-matched groups. Propofol was given intravenously at the speed of 60 ml/min. Encephalographic monitoring was conducted to observe the BIS value. The first administration of propofol was stopped when the BIS value decreased to 80 ~ 75 (Group A), 65 ~ 74 (Group B), 55 ~ 64 (Group C), or < 55 (Group D). The amount of drug administered was recorded. Thirty seconds after the first administration the gastroscope was inserted. If the patients felt uncomforted an additional dose of 30 approximately 50 mg was used. The blood pressure, heart rate, respiratory rate, SpO(2), and BIS value before and during the gastroscopy, times needed for awakening and recovery of orientation were recorded. The patients were asked about if they were conscious during the operation. RESULTS: The doses of propofol in group A, B, C, and D were (1.15 +/- 0.34) mg/kg, (1.43 +/- 0.27) mg/kg, (1.78 +/- 0.31) mg/kg, and (2.15 +/- 0.27) mg/kg respectively, with a significant difference between each 2 groups (all P < 0.05). The successful rate of gastroscope insertion were 17.5%, 47.5%, 92.5%, and 97.5% in group A, B, C, and D respectively, with a. significant difference between each 2 groups (P < 0.05) except between Groups C and D. The heart rate and mean arterial pressure decreased significantly compared with those before the examination in Group D (both P < 0.05). 17 patients of Group A and 4 cases in Group B remembered that they felt unwell during the operation, and 11 patients in Group B said they heard voices but failed to remember the concrete contents. CONCLUSION: It is safe, reliable and feasible to regulate the sedative depth and the dosage of propofol infusion with the BIS monitor. The optimal BIS value in patients undergoing gastroscopy is from 55 to 64. PMID- 17403313 TI - [Upright posture impact on spine susceptibility in scoliosis and progression patterns of scoliotic curve]. AB - OBJECTIVE: To investigate the impact of upright posture on spine susceptibility in scoliosis and scoliotic curve progression patterns. METHODS: There were 3 groups designed in the experiment: bipedal rats group (n = 26), quadrupedal rats group (n = 26) and sham operation bipedal control (n = 12). Taking the advantage of bipedal rats' upright-posture simulating effect, operations were exerted with posterior central approach between scapulas. Along the spatial of erector spinae muscle and trunk muscles, left side upper (T2-3 or T3-4) and lower (T10-11 or 11 12) ribs were tethered with type4 thread to make asymmetrical stress and simulate a primary thoracic curve. At time of 1, 2, 3, 4, 6 and 8 weeks post operation (PO), each group were taken dorsoventral X-ray photographs for dynamic observation on the difference of scoliosis turnover and to compare the curvature progression difference between bipedal and quadrupedal rats. RESULTS: Operation resulted in a primary single thoracic curve or thoracolumbar curve convex to the right in both bipedal and quadrupedal rats. At 1 - 4 weeks PO, all asymmetrical tethers became loose because of rib fracture or osteolysis at the tether sites. All scoliosis of quadrupedal rats disappeared and spine resume straight in 1 - 4 weeks later. Analogically, scoliosis disappeared in bipedal rats for those tethers loose in 1 w PO. But prolonged tethering, scoliosis progression rates increased conspicuously. 3 - 4 weeks' tether time led to irreversible scoliosis. Meanwhile, the curvature progression patterns of bipedal rats presented to be various, including the stable single thoracic curve, progressive thoracolumbar single curve and progressive double curve. These curvatures were much alike that of human idiopathic scoliosis. No scoliosis appeared in sham operation bipedal control. CONCLUSIONS: The upright posture increased spinal susceptibility in scoliosis; Upright posture changes the progression pattern of scoliosis, leading to variegated scoliotic morphouses during primary thoracic curve progression, while inducing curve compensation phenomenon in scoliosis. PMID- 17403314 TI - [The cellular location and significance of p38alpha/beta isoforms in the lumbar spinal cord of the bone cancer pain rats]. AB - OBJECTIVE: To examine the cellular distribution and location of p38alpha/beta isoforms in the lumbar spinal cord of the bone cancer pain rats. METHODS: Twenty SD female rats weighing 180 - 220 g were randomly divided into 2 groups (n = 10 each): group A (control group): intra-tibial injection of 3 microl Hank's solution; group B (model group): intra-tibial injection of 3 microl MADB-106 mammary gland carcinoma cells of rats (4.8 x 10(3)/microl). Mechanical withdrawal threshold and radiant heat threshold of rats' hind paws were measured every other day from one day before operation until 14 days later. The lumbar 4 ~ 5 spinal cord was removed on the 14th day. The cellular distribution and location of the spinal p38alpha/beta immunoreactivity were detected by immunohistochemistry SABC and double immunofluorescence methods. RESULTS: No significant differences in mechanical withdrawal threshold and radiant heat threshold were found at all time points in group A. During the first 6 days of post-operation there was obvious difference in radiant heat stimulus between group A and B (P < 0.05); on the 14th day after operation, mechanical pain threshold and radiant heat threshold in group B were significantly different from that of group A (P < 0.05). The p38alpha/beta immunoreactivity of group B in laminae I approximately IV of dorsal horn showed stronger staining than group A (P < 0.05). Double immunofluorescence confocal micrographs showed that spinal p38alpha and the neuronal marker neuronal N (NeuN) were colocalized in the dorsal horn, indicating that p38alpha was expressed in neurons. Double immunofluorescence micrographs demonstrated that antibodies against p38beta and the microglia marker OX42 labeled the same cell, indicating that p38beta was expressed in microglia. CONCLUSION: Our studies indicate that p38alpha and p38beta are involved in the generation and maintenance of bone cancer pain states. p38alpha is predominantly expressed in neurons, while p38beta is expressed in microglia. PMID- 17403315 TI - [Effect of RNA interference targeting mammalian target of rapamycin on the proliferation of vascular smooth muscle cell and the intimal hyperplasia]. AB - OBJECTIVE: To construct RNA interference (RNAi) vector of mammalian target of rapamycin (mTOR) and study the effect on the proliferation of vascular smooth muscle cell (VSMC) and the intimal hyperplasia (IH). METHODS: The sequence of short hairpin RNA (shRNA) targeting mTOR was designed and synthesized by chemical method and inserted into a retroviral vector pLXIN, then the vector was packaged in PT67 cells. The efficiency of inhibition was verified by Northern blot and Western blot after transfection to VSMC. The changes of p70s 6k and 4E-BP1 were also determined. The proliferation activity of VSMC was determined by flowcytometry and MTT. Autogenous vein graft model were established in 54 rats. Three groups were divided, including gene therapy group, control group and vein graft group. The grafted veins were harvested 7 days, 14 days and 28 days after transplanting respectively. The expression of mTOR was determined by immunohistochemistry stain and Western blot. IH was also observed. The apoptotic VSMC was determined by TUNEL. RESULTS: The shRNA fragments was inserted into pLXIN vector. The shRNA vector was successfully constructed. Additionally, the vector targeting mTOR gene could efficiently decrease the mRNA and protein expression of mTOR and p70s 6k in VSMC, while the expression of 4E-BP1 increased significantly. The cell cycle of VSMC was stunned in phase G(0)/G(1). The protein expression of mTOR in vein graft was inhibited by transfection of pLXIN-shRNA targeting mTOR (P < 0.01) and the IH was also inhibited (P < 0.01), but the apoptosis of VSMC increased significantly (P < 0.01). CONCLUSION: The mTOR RNAi retroviral vector has been constructed successfully, which can significantly inhibit the proliferation of VSMC and the IH, and promote apoptosis after vein grafting. PMID- 17403321 TI - Medicare update: what the latest changes will mean for you. PMID- 17403322 TI - RAPID: rheumatoid arthritis. PMID- 17403323 TI - "Can you call in a Z-Pak for me"? PMID- 17403324 TI - Should HPV vaccination be mandatory? PMID- 17403325 TI - A new papule and "age spots". PMID- 17403326 TI - Time to revise your HIV testing routine. PMID- 17403327 TI - Stress tests: how to make a calculated choice. PMID- 17403328 TI - Should you put all diabetic patients on statins? PMID- 17403329 TI - Clinical inquiries. Do testosterone injections increase libido for elderly hypogonadal patients? PMID- 17403330 TI - Clinical inquiries. How should we treat major depression combined with anxiety? PMID- 17403331 TI - Clinical inquiries. Does warfarin prevent venous thromboembolic events in aPL positive patients? PMID- 17403332 TI - Clinical inquiries. Are there big differences among beta-blockers in treating essential hypertension? PMID- 17403333 TI - Clinical inquiries. What's the best treatment for sebaceous cysts? PMID- 17403334 TI - Clinical inquiries. When (and how) should you evaluate a child for obstructive sleep apnea? PMID- 17403335 TI - [Let's break out of the vicious circle]. PMID- 17403336 TI - [Magnetic resonance imaging of glenohumeral instability: new concepts]. AB - The shoulder is the most unstable joint in the human body and is therefore susceptible not only to acute dislocations but also to minimal states of subluxation related to overuse. The latter condition is known as microinstability and is translated clinically into internal impingement syndromes. Both in dislocations due to acute traumatism and in microinstability, a series of injuries to the glenoid lip, glenohumeral capsule, the articular cartilage, the head of the humerus, the tendons of the rotator cuff, the long tendon of the biceps, the glenohumeral ligaments, and coracohumeral ligament can occur. Glenoid lip lesions are the most common and may present as isolated lesions or in combination with lesions of any of the other structures. These lesions most often affect the anteroinferior margin of the glenoids (the classic Bankard lesion), with different characteristics and variations on its location, (Bankard variants). Another predominant location of glenoid lip lesions is the superior margin (SLAP lesions). Magnetic resonance imaging (MRI), especially after intra articular injection of contrast material, has shown great diagnostic accuracy in this type of lesions, although the interpretation of these images requires knowledge not only of the different lesions and combinations of lesions, but also of the normal variants that can easily be confused with these lesions. PMID- 17403338 TI - [Update on lung cancer staging]. AB - Clinical staging is fundamental in lung cancer, as this descriptive system facilitates the transmission of the extension of the disease, provides prognostic information, helps in choosing the most appropriate treatment program, and provides a standardized system that enables the treatment outcomes of different series of patients to be compared. The most important technique for defining local, regional, and remote extension of the disease continues to be chest and abdominal CT. Functional imaging techniques play a complementary role that is becoming increasingly important in the noninvasive characterization of mediastinal lymph-node involvement. PMID- 17403339 TI - [Ultrasound, CT, and MRI in Crohn's disease]. AB - Colonoscopy and Barium contrast studies continue to be the techniques of choice for the evaluation of mucosal lesions in Crohn's disease. However, these techniques have limited capacity for demonstrating the transmural and extramural extension of the disease. Although ultrasound, CT, and MRI cannot detect early lesions, these techniques satisfactorily show parietal and extraparietal changes caused by the disease, thus complementing the information provided in conventional studies. These three techniques enable the evaluation of the pathological segments, the presence of stenosis and dilatation, and extramural disease (fibrofatty proliferation, fistulae, phlegmons, and abscesses). This information is essential for classifying each patient into one of the three subtypes of Crohn's disease (inflammatory, stenotic, or fistulating). These techniques also enable the evaluation of signs that indicate inflammatory activity (hyperemia, wall enhancement, and increased mesenteric vascularization), and this information, together with the subtype, will determine the treatment. Ultrasound, CT, and MRI should be included in the complete evaluation of the patient with Crohn's disease, together with clinical examination, laboratory tests, and conventional imaging studies. We review the role of ultrasound, CT, and MRI in the diagnosis and evaluation of Crohn's disease, describing patient preparation and examination protocols, as well as the findings in these three techniques. We also discuss the advantages and limitations of each of these techniques and their current clinical roles in this context. PMID- 17403340 TI - [Renal oncocytoma. Fundamental radiologic manifestations and enhancement patterns in tri-phase helical CT]. AB - OBJECTIVE: Oncocytoma is a relatively uncommon benign kidney tumor. To date, it has been impossible to differentiate this tumor from renal cell carcinoma radiologically, although few articles report on the use of tri-phase CT in this tumor. We describe the triphasic CT findings in these tumors and evaluate whether some characteristics, although not sufficient to ensure the diagnosis, can suggest the possibility of oncocytoma. In some cases, this may lead to a different approach to the management of patients. MATERIAL AND METHODS: We describe the tri-phase CT findings in 10 cases of oncocytoma in eight patients (one case was bilateral and multifocal). The diagnosis was made after histological examination of surgical specimens in all cases. RESULTS: All the tumors were found incidentally at ultrasound examination prior to CT study. Tumors all had well-defined borders, and their size ranged from 3 to 15 cm, with a mean diameter of 5.2 cm. One patient had bilateral tumors (2 right and 1 left). All but one of the tumors had a star-shaped scar inside, with a marked lobular pattern in one case. All tumors showed avid uptake, with mean enhancement of 120 HU in the arterial phase and 116 HU in the venous phase. All patients evolved favorably with no post-surgical relapse. CONCLUSIONS: Although oncocytoma cannot be differentiated from renal cell carcinoma with certainty, the possibility of oncocytoma should be suggested in the case of small tumors with a central scar, without necrosis or infiltration, and an enhancement pattern as described here. Regardless of the size of the tumor, lobular morphology should also suggest this possibility. PMID- 17403341 TI - [Multislice CT arteriography in the pre-surgical evaluation of patients with ureteropelvic junction stenosis]. AB - INTRODUCTION: The current treatment of choice for ureteropelvic stenosis is endopyelotomy, but the presence of vessels in contact with the stenotic area of the ureteropelvic junction reduces the success of these interventions and increases vascular and hemorrhagic complications. The aim of our study was to use multislice CT arteriography to evaluate patients prior to surgery for ureteropelvic junction stenosis. PATIENTS AND METHODS: 16 patients with ureteropelvic junction stenosis underwent multislice CT arteriography; multidirectional images and three-dimensional reconstructions were used to identify and characterize vessels in contact with the stenotic area of the ureteropelvic junction. RESULTS: A total of eight vessels (four arteries and four veins) in six (38%) patients were found in contact with or passing within 2 mm of the ureteropelvic junction. The vessels were located anterior to the junction in four cases; it was posterior in one case, and anteromedial in the other. Endopyelotomy was contraindicated in the six patients in whom vessels were found in contact with the ureteropelvic junction. The presence of the vessels was confirmed during surgery in five of these patients; the remaining patient did not undergo surgery). CONCLUSION: Multislice CT arteriography allows adequate presurgical evaluation of ureteropelvic junction stenosis, detecting the presence of crossing vessels and facilitating the planning of the surgical approach in these patients. PMID- 17403342 TI - [Carotid-cavernous fistulas. A report of two cases]. AB - Carotid-cavernous fistulas are uncommon anomalous communications between the venous cavernous sinus and the carotid arterial system. We present two cases studied at our center. We show the characteristic CT, MRI, and orbital ultrasound findings and review the relevant literature. PMID- 17403343 TI - [Primary pancreatic lymphoma in pediatric patients]. AB - Primary involvement of the pancreas in non-Hodgkin's lymphoma is extremely rare in children and adolescents. Indeed, we have only found three cases of primary pancreatic lymphoma in this population published in the English literature. We report the case of a 13-year-old girl diagnosed with primary pancreatic lymphoma. We describe the ultrasonography and computed tomography findings, discuss the differential diagnosis with other pancreatic tumors that affect pediatric patients, and review the literature. PMID- 17403345 TI - [Uterovesical perforation secondary to intrauterine device with vesical lithiasis formation: a report of two cases]. AB - Uterovesical perforation is an uncommon complication in patients with intrauterine devices (IUD). When this complication occurs, intravesical lithiasis is often associated. We present two cases of uterovesical perforation with a description of the radiological findings and a review of the literature. PMID- 17403346 TI - [Preoperative diagnostic imaging in stump appendicitis]. AB - Stump appendicitis is a rare condition in which the diagnosis is often delayed and is eventually reached during exploratory surgery. Knowledge of stump appendicitis makes it possible to reach the diagnosis using imaging techniques and thereby reduce the morbidity of this condition. We present the case of a patient with appendicitis of the appendicular stump that was diagnosed prior to surgery using plain-film radiography and abdominal CT. PMID- 17403351 TI - [Magnetic resonance cholangiography: has it met expectations?]. PMID- 17403352 TI - [Intestinal endometriosis. Current status]. AB - Endometriosis affects a wide spectrum of premenopausal women. Intestinal involvement, affecting mainly the large bowel and sometimes the small bowel, is much less frequent. Diagnosis is relatively straightforward in women with long standing pelvic endometriosis but is difficult in acute intestinal obstruction, since a diagnosis of endometriosis is not often considered in this entity. We performed an exhaustive review of the medical literature, including the option of medical treatment, which is rarely effective in intestinal endometriosis. In most patients with intestinal symptoms, the disease is so severe that surgical treatment is required. Recent studies indicate that the most effective approach is laparoscopic. We analyze the most important classical and recent series of patients and discuss treatment results. PMID- 17403353 TI - [Current status of islet transplantation]. AB - Due to the numerous advances in islet transplantation in the last few years, clinical outcomes following this procedure are continually improving. Novel immunosuppression protocols, improved donor and recipient selection, and careful attention to the process of organ extraction, preservation and islet isolation have contributed to long-term success. The present article reviews the results of clinical islet transplantation and their relationship with the different advances introduced. The use of new islet sources such as living and non-heart-beating donors, as well as recent advances in our knowledge of the mechanisms of rejection and its prevention, are also reviewed. PMID- 17403354 TI - [Laparoscopic vs open splenectomy in the treatment of idiopathic thrombocytopenic purpura]. AB - INTRODUCTION: Laparoscopic splenectomy (LS) is gaining acceptance as an effective and safe alternative to open splenectomy (OS) in the treatment of benign hematologic disorders unresponsive to medical treatment. Among these disorders, the most important is idiopathic thrombocytopenic purpura (ITP). PATIENTS AND METHOD: We compared a cohort of 20 patients who had undergone LS in the previous 7 years, with a historical cohort of 38 patients who underwent OS between 1985 and 1999. All patients in the two groups were diagnosed with ITP unresponsive to medical treatment. RESULTS: The mean age of the patients was 41 years in the LS group and 39.7 in the OS group. Mean spleen size and weight were 11 cm and 150 g in the LS group and 9.9 cm and 190 g in the OS group. Preoperative platelet count was 78 x 10(9) in the LS group and 69 x 10(9) in the OS group. Accessory spleens were detected in 15% of patients in the LS group and in 16% of those in the OS group. The mean operative time was 180 minutes in LS and 85 minutes in OS (p < 0.001). The complications rate was 25% in LS and 21% in OS. The mean length of hospital stay was 3 days in the LS group and 9.4 days in the OS group (p < 0.001). No differences were observed in early and complete long-term remission. CONCLUSIONS: Compared with OS, LS requires a longer operative time and reduces hospital stay. Detection of accessory spleens, complication rates, and effectiveness in terms of early and long-term remission are similar in both procedures. In our opinion, LS should be considered the procedure of choice for the treatment of benign hematological disorders unresponsive to medical therapy. PMID- 17403355 TI - [Laparoscopic adrenalectomy for suspected isolated adrenal metastasis]. AB - INTRODUCTION: Isolated adrenal metastasis is uncommon. Both resection and the laparoscopic approach in this entity are controversial. The aim of this study was to evaluate the indications, diagnosis and utility of laparoscopic adrenalectomy (LA) in patients with isolated adrenal metastasis. PATIENTS AND METHOD: A prospective study was conducted in patients with current or previous tumoral disease and with isolated adrenal metastasis. RESULTS: Sixteen patients, with a mean age of 58 years, were selected. Fifteen LA were performed (one patient was found to have an inoperable tumor at surgery). Histopathological analysis revealed non-tumoral disease in two patients. The most common metastatic disease was non-small cell lung carcinoma (NSCLC) (10 patients), followed by colorectal cancer metastasis (two patients). The mean tumor size was 4.7 cm and was 3.8 cm on computed tomography (p = 0.09). The disease-free interval (DFI) in the NSCLC group was shorter than that in the remaining tumors (p = 0.17). The mean length of follow-up was 39 months, with a mean survival of 39.7 months. The mean actuarial survival at 2 and 5 years was 61% and 17%. At the end of the study, five patients were alive: two were disease free, one had recurrent disease, one had margin involvement and one was awaiting resection of the primary tumor. Eight patients died. One patient survived 9 years after surgery. CONCLUSIONS: LA for metastasis can be performed without oncological disadvantage and should be offered to patients with resectable disease, a DFI > 6 months, and a tumoral size that allows laparoscopic resection. PMID- 17403356 TI - [Adenosquamous and squamous carcinoma of the gallbladder]. AB - INTRODUCTION: Squamous and adenosquamous carcinomas of the gallbladder have poor prognosis. Because these tumors are silent in the initial stage, they are generally diagnosed in advanced stages. MATERIAL AND METHOD: We performed a retrospective observational study of five patients with squamous or adenosquamous carcinoma of the gallbladder. There were two women and three men with a mean age of 71 years (range, 68-75). All patients showed abdominal pain, nausea, vomiting, and weight lost (4-8 kg). One patient presented with acute cholecystitis. None of the patients had jaundice and all had gallstone disease. RESULTS: Surgery was performed in all patients. Extended cholecystectomy was performed in three patients, cholecystectomy and bile drainage in one patient, and colonic bypass and cholecystectomy in the remaining patient. Pathologic analysis revealed epidermoid carcinoma in two patients and adenosquamous carcinoma in three patients. Two patients were treated with adjuvant chemotherapy. All died within 6 months of surgery. CONCLUSIONS: In both histological types of gallbladder carcinoma, treatment depends on the grade of local and regional invasion and tumor spread at diagnosis. PMID- 17403357 TI - [Elective laparoscopic colectomy in uncomplicated diverticulitis: when should surgery be performed]. AB - INTRODUCTION: Surgical resection in acute diverticulitis is indicated after 2-4 episodes, as well as in patients with associated processes. However, the optimal time to perform elective surgery remains to be determined. Compared with open surgery, elective laparoscopic colectomy is associated with fewer postsurgical complications in patients with uncomplicated acute diverticulitis. Nevertheless, the conversion rate to laparotomy is associated with an increase in postoperative morbidity. OBJECTIVE: We studied the impact of time interval to surgery on outcome parameters including operative incidents, postoperative complications and pathologic findings. PATIENTS AND METHOD: Retrospective analysis of two series of case-matched patients according to the timing of operation after the last episode of NCD: group A (within 90 days) and group B (beyond 90 days). Case matching was performed by a computer program according to age, sex, BMI, number of previous episodes, ASA score and prior abdominal surgery. RESULTS: Between July 2000 and June 2004, 132 patients had LCR for NCD. 39 patients were included in group A (median: 40 days, range 21-90 days) vs 38 patients in group B (median: 170 days, range 91-375 days). No patient in either group underwent operation in an emergency setting while waiting for elective surgery. Conversion was required in 5 patients in group A (13%) vs 2 patients in group B (5%) (p = 0.11). Overall abdominal morbidity in Group A was 21% vs 5% in group B (p = 0.02). Mean hospital stay was 7.7 days in group A vs 5.0 days in group B (p = 0.08). Residual inflammation was significantly increased in group A (31%) as compared to group B (11%) (p = 0.01). CONCLUSIONS: Laparoscopic left colonic resection for acute diverticulitis is best performed beyond the third month after the last acute episode. PMID- 17403358 TI - [Acute cholecystitis and laparoscopic cholecystectomy in the elderly]. AB - INTRODUCTION: The aim of this study was to assess the influence of age in laparoscopic cholecystectomy (LC) for the treatment of acute cholecystitis by determining the benefits and postoperative complications in patients older than 65 years. MATERIAL AND METHOD: Between January 2003 and March 2006, we performed 134 urgent LC for acute cholecystitis: 58 patients older than 65 years (group 1) were compared with 76 patients younger than 65 years (group 2). Preoperative, intraoperative and postoperative variables were compared between groups 1 and 2. ASA score was used in the preoperative anesthetic evaluation. RESULTS: A total of 31.2% of patients in group 1 had high surgical risk (24% ASA III and 9.2% ASA IV). The conversion rate was 24.1% in group 1 versus 11.3% in group 2 (p = 0.04), due to difficulty in surgical dissection and advanced cholecystitis. The mean length of postoperative hospital stay was 4.7 +/- 3.2 days in group 1 versus 3.3 +/- 2.4 days in group 2 (p = 0.001). The overall rate of postoperative complications was 33.1% and 18.7% respectively, with a predominance of infectious complications. CONCLUSIONS: Although age should not be an exclusion factor for LC, the conversion rate, postoperative complications and length of hospital stay are increased in the elderly. Higher morbidity due to the underlying disease and longer disease duration with more advanced cholecystitis complicate the laparoscopic approach in these patients. PMID- 17403359 TI - [Curative partial hepatectomy in adult Caroli's disease]. AB - Caroli's disease consists of a congenital malformation of the intrahepatic bile ducts characterized by saccular, segmental, cystic dilatations giving rise to recurrent cholangitis. The inheritance pattern is unclear and the disease may be associated with other hereditary diseases such as adult polycystic kidney disease. The most effective therapeutic option in symptomatic unilobar Caroli's disease is resection of the affected lobe. In bilobar involvement, liver transplantation should be considered. Due to their rarity, we report three cases of Caroli's disease successfully resolved through lobectomy of the affected lobe: two patients with recurrent cholangitis and one patient with associated adult polycystic kidney disease. PMID- 17403360 TI - [High-grade gastric epithelial dysplasia. Diagnostic and therapeutic controversies]. AB - Gastric epithelial dysplasia is a premalignant lesion. Diagnostic and therapeutic criteria are controversial. We report the case of a 52-year-old woman admitted to our hospital. PMID- 17403361 TI - [Diabetic mastopathy. Features in four patients]. AB - Diabetic mastopathy is a clinicopathologic entity that represents less than 1% of benign breast disease and is mainly related to type 1 diabetes. The pathogenesis is unknown but the most convincing hypothesis postulates extracellular collagen deposit and predominantly B-cell inflammation with autoimmune response. The clinical, radiological and pathological features of four patients with diabetic mastopathy are presented. PMID- 17403362 TI - [Direct drainage of segments VI and VII into the gallbladder. Technical implications in cholecystectomy]. PMID- 17403363 TI - [The surgery to come. The feminization of surgery]. PMID- 17403364 TI - [Technical resource for hazardous Kehr T-tube withdrawal]. PMID- 17403365 TI - [Attitude to xenotransplantation in surgical departments in a hospital with a preclinical liver xenotransplant program]. PMID- 17403366 TI - Chylomicrons and lipoprotein lipase at the endothelial surface: bound and GAG ged? AB - At the endothelial cell surface, binding of chylomicrons and lipoprotein lipase (LpL), the major enzyme involved in the processing of these triglyceride-rich lipoproteins, is thought to involve electrostatic interactions with glycosaminoglycans. A new study published in this issue of Cell Metabolism (Beigneux et al., 2007) provides evidence for a specific chylomicron/LpL receptor, which may serve as a platform for LpL-mediated processing of chylomicrons on the capillary endothelium. PMID- 17403367 TI - Oxytocin: the neuropeptide of love reveals some of its secrets. AB - The neuropeptide oxytocin is synthesized in the brain and released from neurohypophyseal terminals into the blood and within defined brain regions that regulate emotional, cognitive, and social behaviors. A recent study of CD38-/- mice (Jin et al., 2007) has demonstrated an essential role for the transmembrane receptor CD38 in secretion of oxytocin into the blood. PMID- 17403368 TI - TOR and aging: less is more. AB - Metabolism and mitochondrial activity are thought to be important determinants of life span. A new study in this issue of Cell Metabolism (Bonawitz et al., 2007) suggests that the TOR pathway controls mitochondrial respiration in yeast and that the harder mitochondria work, the longer yeast live. PMID- 17403369 TI - The GLUT4 glucose transporter. AB - Few physiological parameters are more tightly and acutely regulated in humans than blood glucose concentration. The major cellular mechanism that diminishes blood glucose when carbohydrates are ingested is insulin-stimulated glucose transport into skeletal muscle. Skeletal muscle both stores glucose as glycogen and oxidizes it to produce energy following the transport step. The principal glucose transporter protein that mediates this uptake is GLUT4, which plays a key role in regulating whole body glucose homeostasis. This review focuses on recent advances on the biology of GLUT4. PMID- 17403370 TI - Mitochondrial GTP regulates glucose-stimulated insulin secretion. AB - Nucleotide-specific isoforms of the tricarboxylic acid (TCA) cycle enzyme succinyl-CoA synthetase (SCS) catalyze substrate-level synthesis of mitochondrial GTP (mtGTP) and ATP (mtATP). While mtATP yield from glucose metabolism is coupled with oxidative phosphorylation and can vary, each molecule of glucose metabolized within pancreatic beta cells produces approximately one mtGTP, making mtGTP a potentially important fuel signal. In INS-1 832/13 cells and cultured rat islets, siRNA suppression of the GTP-producing pathway (DeltaSCS-GTP) reduced glucose stimulated insulin secretion (GSIS) by 50%, while suppression of the ATP producing isoform (DeltaSCS-ATP) increased GSIS 2-fold. Insulin secretion correlated with increases in cytosolic calcium, but not with changes in NAD(P)H or the ATP/ADP ratio. These data suggest a role for mtGTP in controlling pancreatic GSIS through modulation of mitochondrial metabolism, possibly involving mitochondrial calcium. Furthermore, in light of its tight coupling to TCA oxidation rates, mtGTP production may serve as an important molecular signal of TCA-cycle activity. PMID- 17403371 TI - Reduced TOR signaling extends chronological life span via increased respiration and upregulation of mitochondrial gene expression. AB - The relationships between mitochondrial respiration, reactive oxygen species (ROS), and life span are complex and remain controversial. Inhibition of the target of rapamycin (TOR) signaling pathway extends life span in several model organisms. We show here that deletion of the TOR1 gene extends chronological life span in Saccharomyces cerevisiae, primarily by increasing mitochondrial respiration via enhanced translation of mtDNA-encoded oxidative phosphorylation complex subunits. Unlike previously reported pathways regulating chronological life span, we demonstrate that deletion of TOR1 delays aging independently of the antioxidant gene SOD2. Furthermore, wild-type and tor1 null strains differ in life span only when respiration competent and grown in normoxia in the presence of glucose. We propose that inhibition of TOR signaling causes derepression of respiration during growth in glucose and that the subsequent increase in mitochondrial oxygen consumption limits intracellular oxygen and ROS-mediated damage during glycolytic growth, leading to lower cellular ROS and extension of chronological life span. PMID- 17403372 TI - Glycosylphosphatidylinositol-anchored high-density lipoprotein-binding protein 1 plays a critical role in the lipolytic processing of chylomicrons. AB - The triglycerides in chylomicrons are hydrolyzed by lipoprotein lipase (LpL) along the luminal surface of the capillaries. However, the endothelial cell molecule that facilitates chylomicron processing by LpL has not yet been defined. Here, we show that glycosylphosphatidylinositol-anchored high-density lipoprotein binding protein 1 (GPIHBP1) plays a critical role in the lipolytic processing of chylomicrons. Gpihbp1-deficient mice exhibit a striking accumulation of chylomicrons in the plasma, even on a low-fat diet, resulting in milky plasma and plasma triglyceride levels as high as 5000 mg/dl. Normally, Gpihbp1 is expressed highly in heart and adipose tissue, the same tissues that express high levels of LpL. In these tissues, GPIHBP1 is located on the luminal face of the capillary endothelium. Expression of GPIHBP1 in cultured cells confers the ability to bind both LpL and chylomicrons. These studies strongly suggest that GPIHBP1 is an important platform for the LpL-mediated processing of chylomicrons in capillaries. PMID- 17403373 TI - Rab10, a target of the AS160 Rab GAP, is required for insulin-stimulated translocation of GLUT4 to the adipocyte plasma membrane. AB - GLUT4 trafficking to the plasma membrane of muscle and fat cells is regulated by insulin. An important component of insulin-regulated GLUT4 distribution is the Akt substrate AS160 rab GTPase-activating protein. Here we show that Rab10 functions as a downstream target of AS160 in the insulin-signaling pathway that regulates GLUT4 translocation in adipocytes. Overexpression of a mutant of Rab10 defective for GTP hydrolysis increased GLUT4 on the surface of basal adipocytes. Rab10 knockdown resulted in an attenuation of insulin-induced GLUT4 redistribution to the plasma membrane and a concomitant 2-fold decrease in GLUT4 exocytosis rate. Re-expression of a wild-type Rab10 restored normal GLUT4 translocation. The basal increase in plasma-membrane GLUT4 due to AS160 knockdown was partially blocked by knocking down Rab10 in the same cells, further indicating that Rab10 is a target of AS160 and a positive regulator of GLUT4 trafficking to the cell surface upon insulin stimulation. PMID- 17403374 TI - Regulation of gluconeogenesis by Kruppel-like factor 15. AB - In the postabsorptive state, certain tissues, including the brain, require glucose as the sole source of energy. After an overnight fast, hepatic glycogen stores are depleted, and gluconeogenesis becomes essential for preventing life threatening hypoglycemia. Mice with a targeted deletion of KLF15, a member of the Kruppel-like family of transcription factors, display severe hypoglycemia after an overnight (18 hr) fast. We provide evidence that defective amino acid catabolism promotes the development of fasting hypoglycemia in KLF15-/- mice by limiting gluconeogenic substrate availability. KLF15-/- liver and skeletal muscle show markedly reduced mRNA expression of amino acid-degrading enzymes. Furthermore, the enzymatic activity of alanine aminotransferase (ALT), which converts the critical gluconeogenic amino acid alanine into pyruvate, is decreased (approximately 50%) in KLF15-/- hepatocytes. Consistent with this observation, intraperitoneal injection of pyruvate, but not alanine, rescues fasting hypoglycemia in KLF15-/- mice. We conclude that KLF15 plays an important role in the regulation of gluconeogenesis. PMID- 17403375 TI - Cytosolic phosphoenolpyruvate carboxykinase does not solely control the rate of hepatic gluconeogenesis in the intact mouse liver. AB - When dietary carbohydrate is unavailable, glucose required to support metabolism in vital tissues is generated via gluconeogenesis in the liver. Expression of phosphoenolpyruvate carboxykinase (PEPCK), commonly considered the control point for liver gluconeogenesis, is normally regulated by circulating hormones to match systemic glucose demand. However, this regulation fails in diabetes. Because other molecular and metabolic factors can also influence gluconeogenesis, the explicit role of PEPCK protein content in the control of gluconeogenesis was unclear. In this study, metabolic control of liver gluconeogenesis was quantified in groups of mice with varying PEPCK protein content. Surprisingly, livers with a 90% reduction in PEPCK content showed only a approximately 40% reduction in gluconeogenic flux, indicating a lower than expected capacity for PEPCK protein content to control gluconeogenesis. However, PEPCK flux correlated tightly with TCA cycle activity, suggesting that under some conditions in mice, PEPCK expression must coordinate with hepatic energy metabolism to control gluconeogenesis. PMID- 17403377 TI - Variability in response to antihypertensive drugs. AB - Heterogeneity of treatment effects (HTE) is a measure of the variations in individual treatment response to the same agent across a population. Hypertension affords an appropriate model for investigators of HTE. Use of blood pressure measurement guidelines and consistent techniques help to reduce the potential variability associated with clinician measurements. Patient characteristics such as age and race/ethnicity can affect blood pressure, including patient response and adverse events observed with antihypertensive medication. Through pharmacogenetic advances, potential underlying causes for such variation are emerging. The growing number of clinical examples of mutations that affect antihypertensive response includes multiple polymorphisms within the components of the renin-angiotensin-aldosterone system. The most prominent examples of these polymorphisms exist in the genes coding for angiotensinogen, angiotensin converting enzyme, and the angiotensin II type 1 receptor. An understanding of the components of blood pressure variability and sources of HTE in antihypertensive therapy is important for analyzing published reports on this topic. It is also helpful when designing treatment protocols for individual patients with hypertension and in assessing their response to therapy. PMID- 17403378 TI - Potential genetic causes of heterogeneity of treatment effects. AB - Nongenetic biologic and lifestyle-related factors, including age, sex, hepatic/renal function, diet/exercise practices, illness severity, smoking, and alcohol consumption habits can account for the heterogeneity of treatment effects (HTE). However, even when these factors are taken into account, considerable variation remains unexplained and could potentially be attributable to genetic differences between patients. Drug response may be dictated by variation in genes involved in both pharmacokinetic (PK) (absorption, distribution, metabolism, excretion [ADME]) and pharmacodynamic (PD) (receptors, ion channels, enzymes, immune system) pathways. Functional variants of the ADME genes can result in patients being poor, intermediate, efficient, or ultrarapid metabolizers of specific agents, thereby affecting efficacy and/or susceptibility to adverse drug reaction and necessitating individualized dosing. A well-documented example of ADME gene variation is the debrisoquine polymorphism, which is characterized by markedly different metabolism of numerous commonly prescribed drugs based on variants of the cytochrome P450 2D6 gene. Variants of genes regulating PD pathways cause altering of drug target pathways, which may affect efficacy in a more pronounced manner. Examples of gene variants affecting PD pathways include those coding for dopamine metabolism, synthesis, and transport. These gene variants may act independently, in combination with each other, and/or in combination with PK genes to affect drug response, for example to antipsychotic medications. Increased understanding of a patient's genotype and its corresponding effect on drug response would be useful to the practicing clinician in choosing an effective drug and in optimizing the dose in a timely manner. PMID- 17403379 TI - Heterogeneity of treatment effects in schizophrenia. AB - The treatment of mental illness presents an opportunity to examine the heterogeneity of treatment effects. The Clinical Antipsychotic Trials of Intervention Effectiveness (CATIE) schizophrenia trial was sponsored by the National Institute of Mental Health (NIMH) to evaluate the effectiveness of antipsychotic medications for schizophrenia in broad patient populations and in scenarios representative of standard clinical practice. Trial inclusion criteria were broad and exclusion criteria were minimal, allowing for a heterogeneous study population. The majority of patients in each phase 1 treatment group discontinued their randomly assigned treatment owing to inadequate efficacy, intolerable side effects, or other reasons. Phase 2 of CATIE featured 2 treatment pathways (efficacy and tolerability) with randomized follow-up medication based on the reason for discontinuation of the previous antipsychotic drug. Outcome differences between treatment groups and variable responses to medications across the study suggest why multiple medication trials are common and may be necessary in the treatment of schizophrenia. Collectively, the CATIE results highlight variable response in the treatment of schizophrenia and demonstrate the need for individualized therapy based on variations in drug efficacy and tolerability among patients. PMID- 17403380 TI - Heterogeneity of treatment effects: implications for guidelines, payment, and quality assessment. AB - Randomized controlled trial results are needed for developing guidelines, payment rules, and quality-of-care measures; however, 2 phenomena reduce the usefulness of randomized controlled trial findings. First, these studies now enroll patients with less severe disease, who are less likely to benefit from a drug or treatment. Second, patients are living longer but, as a result, have more chronic diseases. Although randomized controlled trials often exclude these older patients, trial findings continue to be generalized to them. Together, these phenomena impose challenges to the usefulness of the results of randomized controlled trials for clinical and policy applications. The convergence of these phenomena makes the current research paradigm underlying evidence-based medicine, guideline development and quality assessment fundamentally flawed in 2 ways. First, the "evidence" includes patients who may have a minimal benefit from the treatment being tested. This could reduce the power for the trial and yield negative or null results, leading to undertreatment of a group of patients with potential for a greater-than-observed benefit. Second, attempts to generalize the results from positive trials to patients who have been excluded from those trials may result in the overtreatment of those who could not benefit (e.g., because they will die from other causes before the benefit of treatment would occur) and therefore represents a parallel hazard. In this article, we describe sources of heterogeneity of treatment effects (HTE) within trials, which can compromise the interpretation and generalizability of results. We also examine strategies for understanding and managing HTE in practice, to increase the usefulness of trial results. PMID- 17403381 TI - Healthcare policy implications of heterogeneity of treatment effects. AB - Heterogeneity of treatment effects (HTE) is a phenomenon wherein the same treatment produces different responses in different patients. Following scientific presentations at a conference on HTE in drug therapy, a roundtable panel discussed the policymaking implications of this phenomenon in the current healthcare environment. The presentation of evidence on HTE served as a backdrop for this more pragmatic, solutions-based discussion of how HTE should be addressed in light of the trend in healthcare toward use of evidence-based medicine along with professional society clinical practice guidelines for specific disease states. Overall, the panel concluded that a specific agent should be used when the clinician is equipped with sound data. However, in the absence of such data, care has to be individualized, using the clinician's best judgment regarding available treatment options. The sharing of data across all levels of the healthcare infrastructure is crucial for policymakers seeking to ensure quality care while considering the phenomenon of HTE and, at the same time, keeping cost-effectiveness a major concern. PMID- 17403382 TI - Discussion: 'Risk factors for anal sphincter lacerations' by Lowder et al. PMID- 17403383 TI - Von Willebrand disease presenting as recurrent hemorrhage after transvaginal oocyte retrieval. AB - A 34-year-old nulligravid woman experienced hemorrhage after each of 2 oocyte retrievals. Initial coagulopathy screening was negative. Treatments used during in vitro fertilization likely interfered with assay performance. Reevaluation remote from medications confirmed the diagnosis of von Willebrand disease. Treatments used during in vitro fertilization may increase bleeding risk and confound coagulopathy evaluation. PMID- 17403385 TI - Physician leader should create knowledge-sharing organizational culture. PMID- 17403386 TI - Effects of maternal oxygen administration on fetal pulse oximetry measured by fetal pulse oximetry. PMID- 17403389 TI - Comparing cranial ultrasound results in twin-to-twin transfusion syndrome treated with laser surgery. PMID- 17403390 TI - Hepatic rupture caused by HELLP syndrome? PMID- 17403392 TI - Fetal loss and sickle cell trait: a valid association? [corrected]. PMID- 17403394 TI - Concomitant BRCA1 and BRCA2 gene mutations in an Ashkenazi Jewish woman with primary breast and ovarian cancer. AB - We describe an unusual Ashkenazi Jewish patient with breast and epithelial ovarian cancer who was found to have simultaneous BRCA1 and BRCA2 mutations (BRCA1 5382insC and BRCA2 6174delT). While these mutations are known to occur individually in Ashkenazi Jewish women, concomitant BRCA1 and BRCA2 mutations in the same individual are rare. PMID- 17403396 TI - Preeclampsia and the kidney: footprints in the urine. PMID- 17403397 TI - Progesterone withdrawal: key to parturition. AB - Whereas the essential role of progesterone in the maintenance of pregnancy is accepted generally, the mechanisms that suppress progesterone's function near term to allow labor and delivery of the conceptus are still shrouded in uncertainty. In most subprimate placental mammals, the withdrawal of progesterone before the initiation of labor is manifest by a significant drop in circulating progesterone levels, which is due to either luteolysis or changes in placental steroidogenesis, which shunts precursors towards estrogen production. No such events can be demonstrated in human pregnancy. In this review, we shall present a brief historic background of the research that led to the concepts of "progesterone block" and its withdrawal, based on experiments with rabbits and laboratory rodents, and discuss some of the more recent ideas about "functional progesterone withdrawal," in an attempt to bridge the apparent differences between the regulation of parturition in human and subprimate mammals. PMID- 17403398 TI - Effects of birth spacing on maternal health: a systematic review. AB - The objective of the study was to explore the association between birth spacing and risk of adverse maternal outcomes. The study was a systematic review of observational studies that examined the relationship between interpregnancy or birth intervals and adverse maternal outcomes. Twenty-two studies met the inclusion criteria. Overall, long interpregnancy intervals, possibly longer than 5 years, are independently associated with an increased risk of preeclampsia. There is emerging evidence that women with long interpregnancy intervals are at increased risk for labor dystocia and that short intervals are associated with increased risks of uterine rupture in women attempting a vaginal birth after previous cesarean delivery and uteroplacental bleeding disorders (placental abruption and placenta previa). Less clear is the association between short intervals and other adverse outcomes such as maternal death and anemia. Long interpregnancy intervals are independently associated with an increased risk of preeclampsia. Both short and long interpregnancy intervals seem to be related to other adverse maternal outcomes, but more research is needed. PMID- 17403399 TI - Rapid testing for vaginal yeast detection: a prospective study. AB - OBJECTIVE: The purpose of this study was to determine the accuracy of rapid vaginal yeast detection assay compared with yeast cultures for the diagnosis of vulvovaginal candidiasis. STUDY DESIGN: This was a prospective study that involved 104 subjects, 34 asymptomatic women and 70 symptomatic women with vaginitis. Vaginal swabs were obtained from all subjects for wet mount, yeast culture, and the rapid yeast detection test. Overall, the prevalence rate was 39.4%, based on positive yeast cultures. The rapid yeast test performed by the physician was positive in 30 of 41 subjects with positive cultures and 13 of 63 subjects with negative cultures. RESULTS: The rapid yeast test had 73.1% sensitivity and 82.0% negative predictive value compared with the wet mount, which had 43.9% sensitivity and 70.9% negative predictive value. In symptomatic patients, the test had 77.4% sensitivity and 81% negative predictive value compared with wet mount, which had 51.6% sensitivity. Patient-performed test results were identical to the tests that were performed by the physicians. The cost of the rapid yeast test kit is estimated to be <$10, compared with a mean of $65 for the yeast culture. CONCLUSION: Rapid yeast detection assay is accurate and affordable compared with the gold standard yeast culture in the diagnosis of vulvovaginal candidiasis. Relative to the wet mount, it is more sensitive, cheaper, and accurate for the rapid diagnosis of vaginal yeast infection. PMID- 17403400 TI - Effect of laxatives on gastrointestinal functional recovery in fast-track hysterectomy: a double-blind, placebo-controlled randomized study. AB - OBJECTIVE: The purpose of this study was to determine the effect of early oral bowel stimulation with osmotic laxatives on gastrointestinal function, postoperative nausea and vomiting (PONV) and pain in patients who undergo fast track abdominal hysterectomy. STUDY DESIGN: This was a double-blind, placebo controlled study of 53 women who were assigned randomly to either laxative (magnesium oxide + disodium phosphate) or placebo that was initiated 6 hours after the operation. Primary outcome was time to first defecation; the number of vomiting episodes; nausea and pain score were assessed on a visual analogue scale. RESULTS: Time to first postoperative defecation was a median of 45 hours in the laxative group and a median of 69 hours in the placebo group (P < .0001). There were no significant differences between groups in pain scores, PONV and the use of morphine or antiemetics. Postoperative hospitalization was a median of 1 day in the laxative group and of 2 days in the placebo group (P = .41). CONCLUSION: Laxative improves recovery of gastrointestinal function after fast track hysterectomy but has no significant effect on pain and PONV. PMID- 17403401 TI - Elevated uterine activity increases the risk of fetal acidosis at birth. AB - OBJECTIVE: The objective of the study was to assess the role of uterine activity on fetal outcome. STUDY DESIGN: Intrauterine pressure (IUP) recordings from consecutive term singleton, vaginal deliveries collected between June 1, 1993, and July 1, 2004, were analyzed. One thousand four hundred thirty-three recordings were included. IUP data were obtained using HP 8040A and HP M1350 cardiotocographs. For each recording the uterine contraction curve was analyzed, and the following contraction parameters were determined: relaxation time; contraction duration, frequency, amplitude, and surface; Montevideo units; and active planimeter units and contraction frequency. IUP recordings and contraction parameters from deliveries ending with an umbilical artery pH of 7.11 or less were compared with those ending with an umbilical artery pH of 7.12 or greater. Statistical analyses were performed using Student's t test and logistic regression. RESULTS: An umbilical artery pH 7.11 or less at birth is associated with significant more uterine activity during the first and second stage of labor. CONCLUSION: Increased uterine activity is significantly associated with a higher incidence of an umbilical artery pH of 7.11 or less. PMID- 17403402 TI - Trends in state/population-based Down syndrome screening and invasive prenatal testing with the introduction of first-trimester combined Down syndrome screening, South Australia, 1995-2005. AB - OBJECTIVE: The purpose of this study was to review trends in the us of maternal serum Down syndrome screening and invasive prenatal testing before and after the introduction of a state-based first-trimester combined Down syndrome screening program. STUDY DESIGN: A retrospective population-based study was performed on first- and second-trimester Down syndrome screening, invasive prenatal testing, and prenatal detection of Down syndrome from 1995 to 2005 in South Australia with data from state-based registers. Chi-square tests were used to evaluate trends. RESULTS: There was a significant decrease in the use of second-trimester Down syndrome maternal serum screening (from 75% in 1995 to 25% in 2005; P < .001) and a corresponding significant increase in first-trimester combined screening (from 0.8% in 2000 to 49% in 2005; P < .001). The proportion of all confinements that involved invasive prenatal testing fell (from 9.3% in 1995 to 7.6% in 2005; P < .001). There was a significant decrease in the number of invasive prenatal tests that were needed to detect 1 Down syndrome fetus (from 86 tests in 1995 to 40 tests in 2005; P < .001), with no significant change in the proportion of Down syndrome cases that were detected prenatally. CONCLUSION: The introduction and increased use of first-trimester combined Down syndrome screening has been associated with more efficient use of invasive prenatal testing in South Australia and has maintained a high level of overall prenatal detection. PMID- 17403403 TI - Postpartum headache: is your work-up complete? AB - OBJECTIVE: Headache is a common finding in the postpartum period, and there are limited data describing the cause and treatment of women with postpartum headache. Our objective was to describe our experience with women who were hospitalized for postpartum headache and to develop a management algorithm for these women. STUDY DESIGN: Data for 95 women with headache >24 hours after delivery from 2000-2005 were reviewed retrospectively. Maternal assessment included an evaluation for benign and serious causes of headache that included preeclampsia, dural puncture, and neurologic lesions. Neurologic imaging were performed on the basis of initial neurologic findings and clinical course. Outcomes that were studied included cause, a need for cerebral imaging, neurologic findings, maternal complications, and long-term follow-up evaluations. RESULTS: The mean onset of headache was 3.4 days (range, 2-32 days) after delivery. Tension-type/migraine headache was the most common cause (47%). Preeclampsia/eclampsia and spinal headache comprised 24% and 16% of cases, respectively. Anesthesia evaluation was required in 15 patients because of suspected spinal headache; blood patch was required in 12 of these patients. Cerebral imaging was performed in 22 patients because of focal neurologic deficits and/or failure to respond to initial therapy; 15 of these women (68%) had abnormal findings. Ten patients had serious cerebral pathologic findings, such as hemorrhage, thrombosis, or vasculopathy. There were no deaths; 2 women had minor residual neurologic damage on follow-up evaluation. CONCLUSION: The evaluation of persistent headaches that develop >24 hours after delivery must be performed in a stepwise fashion and requires a multidisciplinary approach. Preeclampsia should be considered initially in women with hypertension and proteinuria. Normotensive women should be evaluated initially for tension type/migraine headache or spinal headache. Patients with headache that is refractory to usual therapy and patients with neurologic deficit require cerebral imaging to detect the presence of life-threatening causes. PMID- 17403404 TI - Urinary podocyte excretion as a marker for preeclampsia. AB - OBJECTIVE: The objective of this study was to examine whether podocyturia, which is the urinary excretion of viable podocytes (glomerular epithelial cells), is present in urinary sediments of patients with preeclampsia. We also aimed to compare the test characteristics of podocyturia to those angiogenic factors that have been shown to play an important role in the pathogenesis of preeclampsia (s Flt-1, PlGF, and endoglin). STUDY DESIGN: Serum angiogenic factors were measured in 44 patients with preeclampsia and 23 normotensive control patients. In a patient subset (15 cases and 16 control patients), urinary proteinuria were identified and quantified on the basis of their expressions of podocyte-specific proteins. RESULTS: Urinary podocyte excretion occurred in all patients with preeclampsia. The positive predictive value for the diagnosis of preeclampsia was greater for podocyturia than for any of the measured angiogenic factors. CONCLUSION: Podocyturia is a highly sensitive and specific marker for preeclampsia. It may contribute to the development of proteinuria in preeclampsia. PMID- 17403405 TI - Gestational weight gain and child adiposity at age 3 years. AB - OBJECTIVE: The purpose of this study was to examine the associations of gestational weight gain with child adiposity. STUDY DESIGN: Using multivariable regression, we studied associations of total gestational weight gain and weight gain according to 1990 Institute of Medicine guidelines with child outcomes among 1044 mother-child pairs in Project Viva. RESULTS: Greater weight gain was associated with higher child body mass index z-score (0.13 units per 5 kg [95% CI, 0.08, 0.19]), sum of subscapular and triceps skinfold thicknesses (0.26 mm [95% CI, 0.02, 0.51]), and systolic blood pressure (0.60 mm Hg [95% CI, 0.06, 1.13]). Compared with inadequate weight gain (0.17 units [95% CI, 0.01, 0.33]), women with adequate or excessive weight gain had children with higher body mass index z-scores (0.47 [95% CI, 0.37, 0.57] and 0.52 [95% CI, 0.44, 0.61], respectively) and risk of overweight (odds ratios, 3.77 [95% CI: 1.38, 10.27] and 4.35 [95% CI: 1.69, 11.24]). CONCLUSION: New recommendations for gestational weight gain may be required in this era of epidemic obesity. PMID- 17403406 TI - Racial/ethnic differences in the monthly variation of preeclampsia incidence. AB - OBJECTIVE: The purpose of this study was to assess monthly patterns in preeclampsia risk separately by race/ethnicity. STUDY DESIGN: We conducted a cohort study of 20,794 white women and 18,916 black women who received care at 12 hospitals in the United States. RESULTS: Among white women, there was a significant U-shaped trend in the incidence of preeclampsia: the incidence was highest in winter months, reached its nadir in mid August, and subsequently increased through the fall months (P < .05). When compared with occurrences in January, the adjusted odds ratios were 0.65 (0.43, 0.99) for May, 0.76 (95% CI, 0.51, 1.13) for June, 0.64 (95% CI, 0.43, 0.97) for July, and 0.64 (95% CI, 0.42, 0.96) for August. Among black women, there was no association between month of delivery and preeclampsia risk (P = .81). CONCLUSION: Our finding that the incidence of preeclampsia decreases in white women during the summer months and that no pattern exists in black women suggests that, in each subgroup, different or competing environmental exposures may be important for the pathogenesis of preeclampsia. PMID- 17403408 TI - Natriuretic peptides and hemodynamics in preeclampsia. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between natriuretic peptides (NT-proANP and NT-proBNP) and hemodynamic parameters in preeclampsia. STUDY DESIGN: This was a cross-sectional study of 19 preeclamptic, 15 chronic hypertensive, and 26 normotensive women in the third trimester of pregnancy. Stroke index (SI), heart rate (HR), cardiac index (CI), systemic vascular resistance index (SVRI), and left cardiac work index (LCWI) were derived by whole-body impedance cardiography. Systolic blood pressure (SAP), diastolic blood pressure (DAP), and mean arterial pressure (MAP) were measured. The plasma levels of NT-proANP and NT-proBNP were determined with radioimmunoassays. RESULTS: NT-proANP and NT-proBNP concentrations were significantly higher in preeclamptic women compared to chronic hypertensive and normotensive pregnancies. Preeclamptic women had lower CI and HR and higher SAP, MAP, and SVRI than the control groups. In preeclampsia NT-proANP correlated significantly with SAP and SVRI; meanwhile, NT-proBNP correlated significantly with SVRI and CI. These correlations persisted in the subgroup of nonmedicated preeclamptic women, except in the case of NT-proBNP and CI. CONCLUSION: High NT-proANP and NT-proBNP concentrations in preeclampsia reflect the strain on the heart caused by high afterload, rather than the function of the heart expressed as SI or CI. PMID- 17403407 TI - Identification of patients at risk for early onset and/or severe preeclampsia with the use of uterine artery Doppler velocimetry and placental growth factor. AB - OBJECTIVE: Preeclampsia has been proposed to be an antiangiogenic state that may be detected by the determination of the concentrations of the soluble vascular endothelial growth factor receptor-1 (sVEGFR-1) and placental growth factor (PlGF) in maternal blood even before the clinical development of the disease. The purpose of this study was to determine the role of the combined use of uterine artery Doppler velocimetry (UADV) and maternal plasma PlGF and sVEGFR-1 concentrations in the second trimester for the identification of patients at risk for severe and/or early onset preeclampsia. STUDY DESIGN: A prospective cohort study was designed to examine the relationship between abnormal UADV and plasma concentrations of PlGF and sVEGFR-1 in 3348 pregnant women. Plasma samples were obtained between 22 and 26 weeks of gestation at the time of ultrasound examination. Abnormal UADV was defined as the presence of bilateral uterine artery notches and/or a mean pulsatility index above the 95th percentile for the gestational age. Maternal plasma PlGF and sVEGFR-1 concentrations were determined with the use of sensitive and specific immunoassays. The primary outcome was the development of early onset preeclampsia (< or = 34 weeks of gestation) and/or severe preeclampsia. Secondary outcomes included preeclampsia, the delivery of a small for gestational age (SGA) neonate without preeclampsia, spontaneous preterm birth at < or = 32 and < or = 35 weeks of gestation, and a composite of severe neonatal morbidity. Contingency tables, chi-square test, receiver operating characteristic curve, and multivariate logistic regression were used for statistical analyses. A probability value of < .05 was considered significant. RESULTS: (1) The prevalence of preeclampsia, severe preeclampsia, and early onset preeclampsia were 3.4% (113/3296), 1.0% (33/3296), and 0.8% (25/3208), respectively. UADV was performed in 95.4% (3146/3296) and maternal plasma PlGF concentrations were determined in 93.5% (3081/3296) of the study population. (2) Abnormal UADV and a maternal plasma PlGF of < 280 pg/mL were independent risk factors for the occurrence of preeclampsia, severe preeclampsia, early onset preeclampsia, and SGA without preeclampsia. (3) Among patients with abnormal UADV, maternal plasma PlGF concentration contributed significantly in the identification of patients destined to develop early onset preeclampsia (area under the curve, 0.80; P < .001) and severe preeclampsia (area under the curve, 0.77; P < .001). (4) In contrast, maternal plasma sVEGFR-1 concentration was of limited use in the prediction of early onset and/or severe preeclampsia. (5) The combination of abnormal UADV and maternal plasma PlGF of < 280 pg/mL was associated with an odds ratio (OR) of 43.8 (95% CI, 18.48-103.89) for the development of early onset preeclampsia, an OR of 37.4 (95% CI, 17.64-79.07) for the development of severe preeclampsia, an OR of 8.6 (95% CI, 5.35-13.74) for the development of preeclampsia, and an OR of 2.7 (95% CI, 1.73-4.26) for the delivery of a SGA neonate in the absence of preeclampsia. CONCLUSION: The combination of abnormal UADV and maternal plasma PlGF concentration of < 280 pg/mL in the second trimester is associated with a high risk for preeclampsia and early onset and/or severe preeclampsia in a low-risk population. Among those with abnormal UADV, a maternal plasma concentration of PlGF of < 280 pg/mL identifies most patients who will experience early onset and/or severe preeclampsia. PMID- 17403409 TI - Protease inhibitor-based antiretroviral therapy and glucose tolerance in pregnancy: AIDS Clinical Trials Group A5084. AB - OBJECTIVE: The objective of the study was to determine whether protease inhibitors increase glucose intolerance and insulin resistance in pregnancy. STUDY DESIGN: In this multicenter, prospective, observational study, 149 human immunodeficiency virus-1-infected pregnant women had fasting insulin, glucose, and C-peptide measured followed by a 1 hour, 50 g glucose test. Glucose intolerance was defined as a 1 hour glucose greater than 130 mg/dL. Glucose intolerance, homeostasis model assessment of insulin resistance and pancreatic beta-cell function, and pregnancy outcomes were compared between those taking protease inhibitors and those not. RESULTS: Fifty-seven of 149 subjects (38%) had glucose intolerance. Body mass index, Hispanic ethnicity, and maternal age, but not protease inhibitors, were associated with glucose intolerance. There were no differences in insulin resistance, beta-cell function, or pregnancy outcome associated with protease inhibitor use. CONCLUSIONS: Protease inhibitors do not increase risk of glucose intolerance or insulin resistance among pregnant women. PMID- 17403410 TI - Combined analysis with amniotic fluid index and estimated fetal weight for prediction of severe macrosomia at birth. AB - OBJECTIVE: To evaluate combined analysis with amniotic fluid index (AFI) and estimated fetal weight (EFW) for prediction of severe macrosomia at birth. STUDY DESIGN: In this retrospective case-control study, 50 term severe macrosomic newborns (birthweight [BW] > or = 97th percentile) were included in the study group and 100 appropriate for gestational age newborns served as controls. All pregnancies underwent a third-trimester sonographic evaluation in which AFI and EFW were measured. The association between BW and AFI and EFW percentiles was examined. The statistical analysis included Student t test, simple regression and receiver-operating curve analyses, and 2x2 tables. RESULTS: The mean mid-third trimester AFI percentile and EFW percentile in severe macrosomic infants were 72.4 +/- 22.5 and 83 +/- 12, respectively, which was significantly higher than in controls (P < .0001). Significant correlations were detected between BW and AFI and EFW percentiles (r = 0.44 and r = 0.72, respectively; P < .0001). Receiver operating characteristic analysis identified AFI > or = 60th percentile and EFW > or = 71st percentile as best predictors of severe macrosomia. The combined analysis with AFI > or = 60th percentile and EFW > or = 71st percentile resulted in a positive predictive value of 85%. CONCLUSION: There is a significant correlation between mid-third-trimester AFI and BW. AFI > or = 60th percentile and EFW > or = 71st percentile during the mid third trimester are useful predictors of severe macrosomia at birth. PMID- 17403411 TI - Hypertensive disorders of pregnancy and the small for gestational age neonate: not a simple relationship. AB - OBJECTIVES: Intrauterine growth restriction (IUGR) reflected by small for gestational age (SGA) status is considered a measure of severity of hypertensive disorders of pregnancy. The purpose of this study was to ascertain whether hypertensive disorders of pregnancy and SGA correlate with severity of hypertension, suggestive of common pathophysiological pathways. STUDY DESIGN: Perinatal and hospital data from 1998-2002 were reviewed. The study identified women with singletons and hypertensive disorders of pregnancy, including those with SGA babies and those with appropriate for gestational age babies. RESULTS: Severity characteristics of hypertensive disorders did not significantly differ between groups (r < .6); however, hypertensive mothers with SGA babies had a higher incidence of a previous SGA baby (but no difference in previous hypertension). CONCLUSION: IUGR in hypertensive disorders of pregnancy is not a measure of severity of hypertension, but reflects underlying fetal susceptibility to growth factors and/or unique (familial) growth patterns. IUGR as a gauge of severity of hypertensive disorders of pregnancy should be reassessed. PMID- 17403412 TI - The association of chronic endometritis with preterm birth. AB - OBJECTIVE: The purpose of this study was to determine whether chronic endometritis (CE) that is diagnosed by endometrial biopsy is associated with preterm birth at <37 weeks of gestation. STUDY DESIGN: Pathology reports for women aged 18-45 years who underwent clinically indicated endometrial biopsy between 1992 and 2002 were solicited from 3 participating clinical pathology laboratories. Reports were dichotomized into those with and those without CE by a standard definition. Results were linked to birth certificates at the Ohio Department of Health. Women who delivered singleton infants (1992-2002) within 3 years of biopsy were included. Biopsy specimens that were obtained from women with conditions that were associated with indicated preterm birth were excluded. Statistical analysis included logistic regression, Student's t-test, Fisher's exact test, and chi-square test, where appropriate. A probability value of <.05 was considered significant. RESULTS: The 1603 endometrial biopsy reports were identified and forwarded. The Ohio Department of Health linked birth certificate data to 193 reports, 157 of which met inclusion criteria and comprised the study group. Twenty-six of 157 women (16.6%) who were studied had CE. The odds of delivering at <37 weeks of gestation were not significantly higher for women with CE (odds ratio, 2.51; 95% CI, 0.86-7.29; P = .091). Infants who were born at <37 weeks of gestation were more likely to be black (P < .001); to have intrapartum fever, meconium, or fetal distress (P = .026) and to require assisted ventilation (P = .016). CONCLUSION: In our study group, women with CE were not more likely to deliver an infant at <37 weeks of gestation. PMID- 17403413 TI - Positive association between a single abnormal glucose tolerance test value in pregnancy and subsequent abnormal glucose tolerance. AB - OBJECTIVE: To study the prevalence of abnormal glucose tolerance among women with a single abnormal glucose tolerance test value in previous pregnancy and identify factors predictive of the later development of abnormal glucose tolerance in this group. STUDY DESIGN: In all, 58 women with gestational diabetes, 66 with a single abnormal value in a glucose tolerance test, and 56 control women underwent a 75-g oral glucose tolerance test at a mean of 6.9 years from the index pregnancy. RESULTS: Abnormal glucose tolerance was present in 34.5% of women with previous gestational diabetes and in 28.7% of women with 1 previous abnormal value, significantly different from the controls (9.7%). Independent risk factors that distinguished the subjects who later developed an abnormal glucose tolerance were prepregnancy BMI, parity > 1, and first-degree relatives affected by diabetes mellitus in the group with gestational diabetes, and prepregnancy BMI, maternal age, (> or = 30 y) and parity > 1 in the group with a single abnormal value. Prepregnancy BMI (> or = 26.9) proved to be the most predictive factor of abnormal glucose tolerance later in life. CONCLUSION: Sicilian women with a single abnormal value at the glucose tolerance test in pregnancy have an increased likelihood of developing an abnormal glucose tolerance later in life, similar to gestational diabetes. Prepregnancy BMI was confirmed as the strongest predictive factor in both groups. PMID- 17403414 TI - Reduced incidence of distant metastases and lower mortality in 1072 patients with breast cancer with a history of hormone replacement therapy. AB - OBJECTIVE: Substitution of estrogens (hormone replacement therapy [HRT]) is the most common therapy and prophylaxis of postmenopausal complaints. However, in most studies, long-term HRT has been associated with an increased risk for breast cancer, but the influence on a prognosis of breast cancer has been examined rarely. STUDY DESIGN: For further investigation, we analyzed 1072 patients aged 45-70 years at the time of first diagnosis of breast cancer with and without preoperative HRT with regard to the incidence of distant metastases and overall survival. Of these, 279 women were premenopausal (mean, 47.8 +/- 3.2 years); 793 women were postmenopausal (mean, 54.5 +/- 3.5 years); 320 women had received HRT over a minimum of 1 year (mean, 5.5 +/- 4.0 years; group HRT+); and 473 women had not received HRT (group HRT-). The median follow-up time was 73.2 months. RESULTS: Although body mass index, tumor size, and grading of group HRT- were significantly higher than in group HRT+, nodal status, S-phase fraction, hormone receptor status, and local recurrence showed no significant differences. In regard to the incidence of distant metastases, women without HRT have significantly (P < .001) more metastases to bone (68 vs 20 women), lung (47:13 women), and liver (47:13 women). Overall survival was significantly lower in the HRT- group. CONCLUSION: We were able to show that the use of HRT before the diagnosis of breast cancer results in more favorable primary tumors, with a lower incidence of recurrences and a better overall survival rate. This might be due to normalized bone metabolism by the use of HRT, which may lower the conditions of tumor cell seeding. PMID- 17403415 TI - Risk factors for primary and subsequent anal sphincter lacerations: a comparison of cohorts by parity and prior mode of delivery. AB - OBJECTIVE: This study was performed to assess the effect of pregnancy, route of delivery, and parity on the risk of primary and subsequent anal sphincter laceration in women at first vaginal delivery (1st VD), vaginal birth after cesarean delivery (VBAC), or second vaginal delivery (2nd VD). METHODS: This retrospective cohort study used data from a perinatal database that included all deliveries at Magee-Womens Hospital from 1995 to 2002. Anal sphincter laceration was the primary outcome, defined as third- and fourth-degree perineal lacerations. The adjusted odds ratio (OR) for primary and subsequent anal sphincter laceration at delivery by risk group was estimated using logistic regression models and reported with 95% confidence intervals (CIs). RESULTS: We assessed 20,674 live, singleton, term deliveries at Magee-Womens Hospital from 1995 to 2002, including 13,183 with 1st VD, 6068 with 2nd VD, and 1423 with VBAC. Anal sphincter laceration occurred in 16% of women with 1st VD, 18% with VBAC, and 3% with 2nd VD. Multivariable logistic regression modeling for primary anal sphincter laceration showed that 1st VD had OR of 5.1 and 95% CI 4.4, 5.9, and VBAC had OR of 5.1, 95% CI 4.2, 6.2 when compared with the reference group with 2nd VD. Shown in order for 1st VD, VBAC, and 2nd VD, the following factors, adjusted for the other listed factors, were significantly related to anal sphincter laceration except as noted: forceps, ORs of 3.0, 2.6, 5.5; midline episiotomy, ORs of 2.7, 2.9, 2.9; infant birth weight 3500 g or more, ORs of 1.9, 1.9, 1.1, not significantly different from OR of 1.0; vacuum delivery, ORs of 1.7, 1.8, 1.5, not significantly different from OR of 1.0, and 2nd stage of labor 2 hours or longer, ORs of 1.8, 0.9, 0.9, last 2 not significantly different from OR of 1.0. Of women who had anal sphincter laceration in their first vaginal delivery, 7.2% (76 of 1054 women who had 2 pregnancies) had recurrent laceration in their second vaginal delivery, compared with 2.3% (123 of 5147) of women who had a primary anal sphincter laceration in their second vaginal delivery. Multivariable logistic regression modeling for recurrent anal sphincter laceration yielded the following significant factors: episiotomy, OR 8.5, 95% CI 4.1, 17.7; vertex malpresentation (primarily occiput posterior), OR 4.3, 95% CI 1.4, 12.6; shoulder dystocia, OR 2.7, 95% CI 1.2, 5.8; and infant birth weight 3500 g or greater, OR 1.7, 95% CI 1.1, 2.7. CONCLUSION: At this institution, women undergoing VBAC are at similarly high risk of anal sphincter laceration, compared with nulliparous women. Women with prior anal sphincter laceration are at 3 times increased risk for subsequent sphincter laceration, compared with women with prior vaginal delivery without sphincter laceration. Pregnancy by itself does not appear to be an important factor in decreasing the risk of anal sphincter laceration in subsequent deliveries. PMID- 17403416 TI - Enhanced synthesis of proinflammatory cytokines by vulvar vestibular fibroblasts: implications for vulvar vestibulitis. AB - OBJECTIVE: The objective of the study was to determine whether vestibular fibroblasts from vulvar vestibulitis (VVS) patients produce higher proinflammatory cytokine levels when provoked with Candida albicans (yeast) and alpha-melanocyte-stimulating hormone (alpha-MSH) in vitro. STUDY DESIGN: Twenty anatomically defined fibroblast strains from patients and age-matched controls were stimulated with 5 regimens: no stimulus, alpha-MSH, heat-killed yeast, alpha MSH plus yeast, and interleukin (IL)-1beta. Supernatant products included the following: granulocyte macrophage colony-stimulating factor, interferon-gamma, IL 10, IL-12, IL-1beta, IL-2, IL-4, IL-6, IL-8, and tumor necrosis factor-alpha were assayed. RESULTS: Coincubation with alpha-MSH plus yeast significantly increased IL-6 (3-fold) and IL-8 (greater than 40-fold) production in patients and controls. Vestibular fibroblast exceeded external vulvar fibroblast production of IL-1beta, IL-6, and IL-8 following yeast alone and alpha-MSH plus yeast stimuli in patients and controls. Substratified by anatomic origin, vestibular fibroblasts from VVS patients produced the highest relative levels of IL-1beta, IL-6, and IL-8 at baseline and following the yeast-alone regimen. CONCLUSION: Localized pain of VVS may results from regionally elevated cytokines produced by vulvar vestibule-specific fibroblasts. PMID- 17403417 TI - Macrophage migration inhibitory factor expression in ovarian cancer. AB - OBJECTIVE: We evaluated the hypothesis that ovarian cancer patients have significantly higher levels of serum macrophage migration inhibitory factor (MIF). STUDY DESIGN: MIF levels were determined by enzyme-linked immunosorbent assay (ELISA) in epithelial ovarian cancer cell lines and immortalized normal ovarian surface epithelial cells and in serum of ovarian cancer patients (n = 54) and age-matched healthy women (n = 60). To determine the impact of Toll-like receptor-4 ligation on MIF levels, cells were treated for 48 hours with lipopolysaccharide. RESULTS: Cancer cells, but not normal cells, secrete significant amounts of MIF. This correlates in vivo, where serum MIF levels are significantly higher in ovarian cancer patients. Treatment of cancer cells with lipopolysaccharide induced a significant increase in MIF secretion. CONCLUSION: MIF may be relevant in the process of ovarian cancer formation and progression. The events leading to the induction of MIF expression and its contribution to ovarian cancer progression may open new venues for targeted therapy. PMID- 17403418 TI - Regulation of MMP-1 by sex steroid hormones in fibroblasts derived from the female pelvic floor. AB - OBJECTIVE: The aim of this study was to investigate the effect of sex steroid hormones on the overall expression profile of cellular matrix metalloproteinase-1 (MMP-1) in fibroblasts that are derived from arcus tendineus fasciae pelvis. STUDY DESIGN: Arcus tendineus fasciae pelvis fibroblasts that originated from a premenopausal woman and a postmenopausal woman who was undergoing a prolapse repair were treated with physiologic concentrations of 17-beta-estradiol (E2), progesterone, E2 plus progesterone, and E2 plus ICI 182,780. Cellular expressions of the latent, active, and fragment forms of MMP-1 were analyzed quantitatively by Western immunoblotting. RESULTS: The latent and fragment forms of MMP-1 were increased by E2, progesterone, and E2 plus progesterone. The active form of MMP-1 was not changed by either E2 or progesterone alone but was decreased significantly when both hormones were added together. ICI 182,780 inhibited the stimulatory effect of E2. CONCLUSION: Fragmentation is a site of regulation of MMP-1 expression by hormones. Only E2 combined with progesterone decreased the active form of MMP-1, which suggests that both hormones are necessary to maintain the integrity of female pelvic floor. PMID- 17403419 TI - Fetal gene transfer with lentiviral vectors: long-term in vivo follow-up evaluation in a rat model. AB - OBJECTIVE: The purpose of this study was to evaluate the long-term expression of a transgene and subsequent immune response after the injection of lentiviral vectors in a fetal rats. STUDY DESIGN: Fetal rats were injected in the liver, peritoneal cavity, or lung at E19 (term, E21) with a lentiviral vector expressing enhanced green fluorescent protein and luciferase. Controls received saline solution. After birth, full body bioluminescence was done at weeks 1, 4, 10, and 30 of life; seroconversion for the transgene was assessed. RESULTS: All surviving fetuses that had been injected in the liver (8/9 fetuses), peritoneum (3/3 fetuses), or lung (9/10 fetuses) showed a signal on bioluminescence imaging scan up to 30 weeks. None of the survivors displayed seroconversion against the transgene. CONCLUSION: In the rat model, the administration of lentiviral vectors into the fetal lung and liver resulted in long-term transgene expression without detectable humoral immune response. PMID- 17403420 TI - The effect of the postnatal environment on altered fetal programming of adult vascular function in mice that lack endothelial nitric oxide synthase. AB - OBJECTIVE: The purpose of this study was to investigate vascular reactivity in heterozygous and homozygous offspring with a genetic predisposition for hypertension after postnatal cross-fostering to mothers with the opposite genetic inheritance of the NOS3 knockout allele. STUDY DESIGN: Homozygous NOS3 knockout (C57BL/6J-NOS3(-/-KO)) and wild-type mice (NOS3(+/+WT)) were bred to obtain heterozygous litters with a paternally derived (NOS3(+/-pat)) or maternally derived (NOS3(+/-mat)) knockout allele. After delivery, heterozygous and homozygous litters were cross-fostered to a mother with the opposite NOS3 gene status. Carotid arteries were placed in a wire myograph for isometric tension recording with the use of contractile and relaxant agents. Statistical analysis with 1-way analysis of variance and Neuman-Keuls post-hoc testing was performed. RESULTS: Increased sensitivity to phenylephrine and absent relaxation to acetylcholine in NOS3(+/-mat) was reversed with cross-fostering, and vasorelaxation to isoproterenol was increased. Contraction to calcium was increased in the cross-fostered paternally derived and wild-type litters. CONCLUSION: Postnatal interventions may alter the adult vascular profile favorably that is the result of an abnormal intrauterine environment. PMID- 17403421 TI - Corticotropin-releasing factor inhibition of sheep fetal colonic contractility: mechanisms to prevent meconium passage in utero. AB - OBJECTIVE: In humans, fetal in utero meconium (MEC) passage rarely occurs before term gestation. We hypothesized the existence of inhibitory mechanism(s) preventing colonic motility and MEC passage prior to term. STUDY DESIGN: Longitudinal smooth muscle strips prepared from distal colon of preterm ovine fetuses (130-132 d; term = 148-152 d) were examined for their contractile responses to muscarinic receptor agonist (bethanechol) and both nonspecific (atropine) and receptor subtype specific antagonists (M1: pirenzepine dihydrochloride, M2 methoctramine, M3: 4-diphenylacetoxy-N-methlpiperidine methiodide [4-DAMP] and M4: tropicamide) in an in vitro organ bath system. Effects of corticotrophin releasing factor (CRF) and Urocortin I (URO-I), known modulators of colonic motility and smooth muscle contractility, were studied on bethanechol-induced contractility. Immunohistochemical analysis was performed to confirm the expression of CRF and URO-I, and muscarinic and CRF R2 receptors in distal colon. RESULTS: Bethanechol induced smooth muscle contractions via muscarinic receptor subtype M3. CRF and URO-I elicited a significant inhibition of bethanechol induced contraction. Immunohistochemical analysis verified the expression of muscarinic receptor subtype M3, CRF, URO-I and CRF-receptor-R2 in distal colon. CONCLUSION: Inhibition of M3 dependent distal colonic motility by CRF system may prevent the passage of MEC in the preterm ovine fetus. PMID- 17403422 TI - Fetal cerebral oxygenation: the role of maternal hyperoxia with supplemental CO2 in sheep. AB - OBJECTIVE: We tested the hypothesis that supplemental CO2 can enhance the effect of maternal oxygen administration on fetal cerebral oxygenation. STUDY DESIGN: In near-term fetal sheep (n = 6), we instrumented the cerebral cortex with tissue PO2-laser Doppler flow probes, and placed arterial catheters. Following a 30 minute control period, the ewe breathed 50% O2 for 15 minutes, followed by added 6% CO2 for 15 minutes. We examined fetal cortical tissue PO2, cerebral blood flow (CBF), and fetal and maternal blood gases and related variables. RESULTS: In response to maternal O2 administration, fetal arterial PO2, O2 content, cerebral O2 delivery, and cortical tissue PO2 increased significantly. In response to supplemental CO2 inhalation, fetal cortical tissue PO2 increased further. Fetal CBF also increased in response to the elevated arterial CO2 level. CONCLUSION: CO2 supplementation of maternal O2 administration enhanced fetal cerebral oxygenation. In contrast, it was considered that during labor maternal hyperventilation with hypocapnia may blunt the effect of maternal O2 inhalation. PMID- 17403423 TI - Prostaglandin E2 synthase in syncytiotrophoblastic vesicles found in the placental intervillous space. AB - OBJECTIVE: The purpose of this study was to confirm microsomal prostaglandin E2 synthase (mPGES) in the vesicles found in the intervillous space. STUDY DESIGN: Immunohistochemical staining for mPGES was conducted to confirm this enzyme in vesicles by using fresh sections of mature placentas. RESULTS: Staining for mPGES was prominent in vesicles separating from syncytiotrophoblast into the intervillous space. Syncytiotrophoblast was partly positive or negative for mPGES. Vesicles showed a tendency to degenerate with a weakly positive or negative reaction for mPGES. Extravillous cytotrophoblast was strongly positive for mPGES in the basal plate. CONCLUSION: The vesicles containing mPGES suggest the formation of prostaglandin E2 (PGE2). This enzyme could be concentrated in vesicles by a different secretory mechanism. PMID- 17403424 TI - Usefulness of a placental profile in high-risk pregnancies. AB - OBJECTIVE: Test the hypothesis that a placental function profile can reassure most high-risk women with normal test results yet accurately can identify a subset of women who are destined for major complications that will be attributable to placental disease. STUDY DESIGN: This was a prospective study of 212 high-risk pregnancies that used the placental profile (16- to 18-week maternal serum screening, 18- to 23-week uterine artery Doppler imaging, and placental morphologic condition). Odds ratios (95% CI) were derived for intrauterine fetal death (IUFD), preterm delivery at < 34 weeks of gestation, preeclampsia/hemolysis, elevated liver enzymes, and low platelet count syndrome (HELLP) syndrome, small for gestational age delivery, and early-onset intrauterine growth restriction (IUGR); all normal test results (n =125) were compared with > or = 1 abnormal test results. RESULTS: The odds of the development of adverse outcomes were significantly less in women with all normal test results (preeclampsia/HELLP [odds ratio, 0.2; 95% CI, 0.1-0.4]), preterm delivery (odds ratio, 0.1; 95% CI, 0.06-0.3), small for gestational age delivery (odds ratio, 0.2; 95% CI, 0.09-0.3), early-onset IUGR (0), and IUFD (odds ratio, 0.05 [0.01-0.2]). Combining those women with two (n = 21) of 3 (n = 15) abnormal test results together predicted 14 of 19 severe IUGR and 15 of 22 IUFD cases. CONCLUSION: This placental function profile at 16-23 weeks of gestation can reassure women with normal test results by identifying a smaller subset of women who are at reduced risk of perinatal morbidity or death from severe IUGR. PMID- 17403425 TI - The pursuit of evidence. PMID- 17403426 TI - Genetic communication between fetus and mother: short- and long-term consequences. PMID- 17403427 TI - Progestin inhibits and thrombin stimulates the plasminogen activator/inhibitor system in term decidual stromal cells: implications for parturition. AB - OBJECTIVE: Labor is associated with 'decidual activation' with increased proteolysis and extracellular matrix degradation. The balance between plasminogen activator inhibitor-1 (PAI-1) and urokinase (uPA) and tissue-type plasminogen activator (tPA) is an important determinant of proteolytic activity at the maternal-fetal interface. Thrombin released at the time of placental abruption (decidual hemorrhage) is known to promote decidual proteolysis and uterine contractions. This study investigates the separate and interactive effects of steroid hormones and thrombin on PAI-1, uPA, and tPA expression by term decidual cells (DCs). STUDY DESIGN: Term DCs were isolated by enzymatic digestion, purified, and depleted of leukocytes. Cells were treated with estradiol (10(-8) mol [E2]), medroxyprogesterone acetate (10(-7) mol [MPA]), both, or vehicle for 7 days. After 24-hour incubation with or without thrombin (0.1-2.5 U/mL), levels of PAI-1, uPA, and tPA in conditioned supernatant were measured by specific ELISA and Western blotting. Levels of PAI-1 and uPA mRNA were measured by quantitative RT-PCR. RESULTS: In the cultured term DCs, ELISA measurements indicated that basal output of PAI-1 was about 2 logs higher than that of either uPA or tPA (2.5 +/- 0.7 ng/mL per microg protein, 13.4 +/- 6.3 pg/mL per microg protein, and 25.4 +/- 10.8 pg/mL per microg protein, respectively). Although E2 alone did not affect PAI-1 output, MPA and E2+MPA significantly enhanced PAI-1 production (2.5 +/- 0.7 vs 8.2 +/- 2.0 ng/mL per microg protein for E2+MPA [3.3-fold]; P < .01). By contrast, uPA output was inhibited by exposure to MPA (13.4 +/- 6.3 vs 2.6 +/- 1.1 pg/mL per microg protein [0.2-fold]; P < .05), whereas tPA production was not affected by MPA. Thrombin did not significantly affect uPA and tPA production by term DCs. In contrast, in E2+MPA-treated term DCs, thrombin, a hemostatic proinflammatory cytokine, selectively increased PAI-1 output in a dose-dependent fashion, which could be blocked by the selective thrombin inhibitor, hirudin. Western blotting confirmed the effects of MPA and thrombin in elevating secreted levels of PAI-1. Unlike the increase in PAI-1 output elicited by thrombin, term DCs were unresponsive to either of the classic proinflammatory cytokines, TNFalpha or IL-1beta. Corresponding effects on PAI-1 mRNA levels were elicited by MPA and thrombin as seen for PAI-1 protein expression, suggesting that these up regulatory effects are transcriptionally mediated. CONCLUSION: Progestin enhanced PAI-1 and inhibited uPA expression by term DCs, which may explain in part the pregnancy-prolonging properties of progesterone as a consequence of inhibited proteolytic activity at the maternal-fetal interface. Thrombin augmented PAI-1 expression in the absence of increased uPA or tPA expression by term DCs, suggesting that abruption-associated decidual proteolysis and preterm labor is mediated primarily by thrombin-enhanced matrix metalloproteinase expression rather than an indirect effect on the plasminogen activator/inhibitor system. PMID- 17403428 TI - Recurrent spontaneous abortion and skewed X-inactivation: is there an association? AB - OBJECTIVE: The purpose of this study was to determine whether there is an association between skewed X-inactivation and recurrent spontaneous abortion in a large, well-defined sample of women with recurrent loss. STUDY DESIGN: X chromosome inactivation patterns were compared in 5 groups of women. Group 1 (recurrent spontaneous abortion) consisted of 357 women with 2 or more spontaneous losses. In group 2 (infertility), there were 349 subjects from infertility practices recruited at the time of a positive serum beta-human chorionic gonadotropin. Group 3 (spontaneous abortion) women (n = 81) were recruited at the time of an ultrasound diagnosis of an embryonic demise or an anembryonic gestation. Groups 4 (primiparous) and 5 (multiparous) were healthy pregnant subjects previously enrolled in another study to determine the incidence and cause of pregnancy complications, such as preeclampsia and intrauterine growth restriction. The Primiparous group included 114 women in their first pregnancy, whereas the Multiparous group consisted of 79 women with 2 or more pregnancies but without pregnancy loss. RESULTS: The rate of extreme skewing (90% or greater) in the recurrent spontaneous abortion population was 8.6%, and not statistically different from any of the other groups, except the Primiparous group (1.0%, P < .01). The incidence of X-inactivation skewing of 90% or greater was no different whether there had been at least 1 live birth (9.9%), or no previous live births and at least 3 losses (5.6%, P > .05). When age and skewing of 90% or greater are compared, subjects with extreme skewing have a mean age of 2 years older than those without extreme skewing (P < .05). CONCLUSION: Skewed X inactivation is not associated with recurrent spontaneous abortion but is associated with increasing maternal age. PMID- 17403429 TI - GPR30: a novel indicator of poor survival for endometrial carcinoma. AB - OBJECTIVE: This study was undertaken to evaluate the relationship between GPR30, classical steroidal receptor expression, and clinical outcome in patients with endometrial carcinoma. STUDY DESIGN: Immunohistochemistry was used to investigate the expression of GPR30, estrogen, progesterone, epidermal growth factor receptors and Ki-67 in 47 consecutive consenting patients with endometrial carcinoma diagnosed between 1997 and 2001. Results were correlated with clinical and pathologic predictors of adverse outcome and survival. RESULTS: GPR30 correlated positively with epidermal growth factor receptor (P = .005), but negatively with progesterone (P = .05) receptor expression. GPR30 overexpression occurred more frequently in tumors with deep myometrial invasion, high-grade, biologically aggressive histologic subtypes, and advanced stage. In patients with GPR30 overexpression, survival was significantly poorer (65.2% vs 100%, P = .005). CONCLUSION: GPR30 represents an alternative estrogen-responsive receptor that is overexpressed in tumors where estrogen and progesterone receptors are downregulated, and in high-risk endometrial cancer patients with lower survival rates. PMID- 17403430 TI - Identifying the safety profile of a novel infectivity-enhanced conditionally replicative adenovirus, Ad5-delta24-RGD, in anticipation of a phase I trial for recurrent ovarian cancer. AB - OBJECTIVE: The purpose of this study was to evaluate the biodistribution and toxicity of the tropism-modified infectivity-enhanced conditionally replicative adenovirus, Ad5-delta24-arginine-glycine-aspartate (RGD). STUDY DESIGN: Cohorts of cotton rats were treated intravenously or intraperitoneally for 3 consecutive days with 5 x 10(8) to 5 x 10(11) particles/kg of Ad5-delta24-RGD or controls and killed on day 8, 17, or 56. For biodistribution studies, tissue samples from 14 organ sites and serum samples were evaluated for the presence of virus with the use of quantitative polymerase chain reaction analysis. For toxicity experiments, tissue samples from more than 30 organ sites and serum samples were obtained for the assessment of vector-related tissue or laboratory effects. RESULTS: Ad5 delta24-RGD was noted in tested samples at days 8 and 17 in animals that were treated intravenously and intraperitoneally with clearance by day 56. There were lower copies of vector noted in the blood and liver specimens of intraperitoneally treated animals. Mild peritonitis histopathologic findings were noted in rats that were treated intraperitoneally with Ad5-delta24-RGD; pathologic findings did not vary significantly with dose, over time, or in comparison to that noted in animals that were treated with Ad5-delta24. CONCLUSION: These studies provide critical insights regarding Ad5-delta24-RGD dosing and anticipated toxicity for a planned clinical trial for ovarian cancer. PMID- 17403431 TI - Stromal cells of endometriosis fail to produce paracrine factors that induce epithelial 17beta-hydroxysteroid dehydrogenase type 2 gene and its transcriptional regulator Sp1: a mechanism for defective estradiol metabolism. AB - OBJECTIVE: In endometrium, stromal progesterone receptors mediate production of paracrine factors, which enhance binding of the transcription factor specific protein-1 to the promoter of the gene encoding the 17beta-hydroxysteroid dehydrogenase type 2 enzyme responsible for converting biologically active estradiol to estrone in epithelium. The objective of this study is to define the cellular defect responsible for the disruption of this stromal-epithelial interaction in endometriosis. STUDY DESIGN: We determined the effects of conditioned media generated from primary human eutopic endometrial stromal cells vs endometriotic stromal cells on Ishikawa malignant endometrial epithelial cells. RESULTS: Conditioned media from progestin-pretreated eutopic endometrial stromal cells but not endometriotic stromal cells significantly stimulated specific protein-1 protein levels, 17beta-hydroxysteroid dehydrogenase type 2 messenger RNA levels and promoter activity, and binding activity of specific protein-1 to the 17beta-hydroxysteroid dehydrogenase type 2 promoter region in Ishikawa cells. CONCLUSION: A stromal cell defect in endometriosis blocks formation of progesterone-dependent production of factors leading to 17beta hydroxysteroid dehydrogenase type 2 deficiency and defective conversion of estradiol to estrone in epithelium. PMID- 17403432 TI - Identification of proteins within the nuclear factor-kappa B transcriptional complex including estrogen receptor-alpha. AB - OBJECTIVE: The objective of the study was to determine whether cross-talk occurs between estrogen receptors (ERs) and nuclear factor-kappa-B (NF-kappaB), to assess the functional consequences of such an ER/NF-kappaB interaction, and to identify other unknown regulatory proteins that may participate in the NF-kappaB transcriptional complex. STUDY DESIGN: Electromobility gel shifts, reporter gene assays, and mass spectrometry were used to identify proteins interacting with the NF-kappaB deoxyribonucleic acid (DNA) response element. RESULTS: ER and the p65 subunit of NF-kappaB colocalized on DNA. This interaction was inhibitory for ER transcriptional activity. Sequencing of proteins bound to the NF-kappaB/DNA complex identified DNA-modifying enzymes, scaffolding proteins, chaperones, and elements of the nuclear matrix. CONCLUSION: These studies have identified an inhibitory interaction between estrogen receptors and the p65 subunit of NF kappaB with implications for estrogen action in pregnancy and cancer. New accessory proteins have also been identified that bind to protein complexes on the NF-kappaB DNA response element. PMID- 17403433 TI - The effect of over-expression of sFlt-1 on blood pressure and the occurrence of other manifestations of preeclampsia in unrestrained conscious pregnant mice. AB - OBJECTIVE: It has been shown that the level of soluble fms-like tyrosine kinase-1 (sFlt-1) is elevated in pregnant women who are destined to have preeclampsia, and a role for sFlt-1 in its pathogenesis has been suggested. Our objective was to determine the effect of the over-expression of sFlt-1 on blood pressure and the occurrence of other manifestations of preeclampsia in pregnant mice. STUDY DESIGN: At day 8 of gestation CD-1 mice were allocated randomly to an injection of an adenovirus carrying sFlt-1 (10(9) plaque-forming units; sFlt-1 group), adenovirus carrying the murine immunoglobulin G2alpha Fc fragment (10(9) plaque forming units; mFc group used as a control for the virus) or saline solution (100 microL; saline group). At day 10 of gestation, blood pressure catheters were inserted through the left carotid artery into the aortic arch and tunneled to a telemetric transmitter. Blood pressure was monitored continuously in the conscious unrestrained animals until day 18. Blood was collected from the pregnant mice at different gestational times, and plasma sFlt-1 was measured by enzyme-linked immunosorbent assay. Pups and placentae were weighed, and maternal platelet counts were determined at death on day 18. RESULTS: Plasma levels of sFlt-1 increased significantly in the sFlt-1 mice and were significantly higher than the 2 control groups. The mean blood pressure in the sFlt-1 mice was significantly higher on days 17 and 18 of gestation, compared with the mFc and saline solution groups. The time-course of blood pressure rise mirrored that of the sFlt-1 levels. The average pup weight, placental weight, and maternal platelet counts were significantly lower in the sFlt-1 group, compared with the controls. CONCLUSION: SFlt-1 induces hypertension and fetal growth restriction in pregnant mice, which supports its hypothesized role in the pathogenesis of preeclampsia. This animal model minimizes the need for manipulation or the administration of various compounds to induce the condition. PMID- 17403434 TI - Correlation of adrenocorticotropin steroid levels between women with polycystic ovary syndrome and their sisters. AB - OBJECTIVE: The purpose of this study was to assess the sibling correlation of adrenocorticotropic hormone-stimulated steroid hormone levels between probands with polycystic ovary syndrome and their sisters. STUDY DESIGN: Twenty-seven women with polycystic ovary syndrome and 28 of their sisters underwent an adrenocorticotropic hormone stimulation test with measurement of the steroids dehydroepiandrosterone, androstenedione, and cortisol 60 minutes later. The 60 minute values were used to calculate sister-sister correlations by regression analyses. RESULTS: The adrenocorticotropic hormone-stimulated (60-minute) log transformed dehydroepiandrosterone and cortisol values were significantly correlated between siblings (r = 0.47, P = .01 and r = 0.57, P = .01, respectively); a similar trend was observed for the 60-minute A4 values (r = 0.29, P = .06). CONCLUSION: Women with polycystic ovary syndrome and their sisters have significantly correlated levels of adrenocorticotropic hormone stimulated steroids, supporting a genetic basis of the adrenal androgen excess observed in polycystic ovary syndrome. PMID- 17403435 TI - Increased leukocyte adhesion to vascular endothelium in preeclampsia is inhibited by antioxidants. AB - OBJECTIVE: To test the hypothesis that plasma from women with preeclampsia increases leukocyte adhesion to vascular endothelial cells and that antioxidants inhibit this effect. STUDY DESIGN: Plasma from 12 women with severe preeclampsia and 12 with normal pregnancy was tested in an in vitro leukocyte-endothelium adhesion assay in the presence or absence of vitamin E, vitamin C, or N acetylcysteine. RESULTS: Preeclamptic plasma significantly increased monocyte (U937 cells) and T-cell (Jurkat) adhesion to human umbilical vein (HUVEC) and microvascular endothelial cells, compared with normal pregnant plasma. The antioxidants vitamin E, vitamin C, and N-acetylcysteine significantly inhibited monocyte adhesion to HUVEC in the presence of preeclamptic but not normal pregnant plasma. Increased adhesion in response to preeclamptic plasma was not mediated through a protein kinase C (PKC) mechanism, because the PKC inhibitor bisindolylmaleimide I had no effect on adhesion in the presence of preeclamptic plasma. CONCLUSION: Severe preeclampsia is associated with increased leukocyte endothelium adhesion and clinically useful antioxidants can inhibit this effect. PMID- 17403436 TI - The effects of metformin and rosiglitazone, alone and in combination, on the ovary and endometrium in polycystic ovary syndrome. AB - OBJECTIVE: To examine the effects of metformin and rosiglitazone, alone and in combination, on endometrial histology and ovarian steroid production. STUDY DESIGN: Randomized open-label study of metformin and rosiglitazone in 16 women with polycystic ovary syndrome (PCOS) performed at a single academic health center. The study consisted of a 6-week baseline observation period, a 3-month treatment period of single-agent therapy (rosiglitazone or metformin), and then a 3-month period of combined therapy. RESULTS: Abnormal endometrial histology was found in 3 subjects at baseline, including 1 case of adenocarcinoma of the endometrium in an asymptomatic subject, who was excluded from further study. The 2 other abnormal cases (simple hyperplasia) resolved with treatment. Three months of single-agent therapy showed a benefit of rosiglitazone (n = 9) over metformin (n = 6) in terms of reducing circulating unbound testosterone levels (-11.8; 95% CI: -21.7 to -2.0 ng/dL) and 2-hour glucose (-42.0; 95% CI: -76.2 to -7.8 mg/dL), 2-hour insulin (-150.4; 95% CI: -272.7 to -28.1 microU/mL) as well as a significant decrease in integrated levels of glucose and insulin by area under the curve analysis, all obtained from oral glucose tolerance testing. Daily urinary progestin-to-estrogen ratios improved on rosiglitazone compared to metformin therapy (0.08; 95% CI: 0.02 to 0.14). Ovulatory rates tended to improve on both single-agent and combined treatments (30/90 cycles, 33%), compared to baseline ovulatory rate (2/15, 13%). Despite 6 months of therapy alone or in combination, 5 women displayed no evidence of biochemical ovulation by urinary or serum progestin measurements. CONCLUSION: This study provides preliminary evidence that insulin-sensitizing drugs may have beneficial effects on the endometrium, although the exact mechanism beyond improving ovulatory function is still unknown. In addition, we suggest that rosiglitazone may be more beneficial than metformin therapy on raised insulin and androgen levels in an obese PCOS population. Combined therapy did not demonstrate significant benefit above and beyond single-agent therapy. PMID- 17403437 TI - The PESSRI study: symptom relief outcomes of a randomized crossover trial of the ring and Gellhorn pessaries. AB - OBJECTIVE: The aim of this randomized crossover trial was to compare symptom relief and change in life impact for women using the ring with support and Gellhorn pessaries. STUDY DESIGN: Subjects were randomized to use each pessary for 3 months. Outcome data included a visual analog satisfaction score, and quality of life questionnaires. Analysis included student's t-test, Wilcoxan Signed-rank test and logistical regression. RESULTS: Subjects were primarily white, parous, postmenopausal women with a mean age of 61. The median POPQ stage was III. We enrolled 134 subjects and collected 3-month data on 94 ring and 99 Gellhorn subjects. There were statistically and clinically significant improvements in the majority of the PFDI and many PFIQ scales with both pessaries, but no clinically significant differences between the two pessaries. CONCLUSIONS: The ring with support and Gellhorn pessaries are effective and equivalent in relieving symptoms of protrusion and voiding dysfunction. PMID- 17403438 TI - Cervical cancer prevention: safety, acceptability, and feasibility of a single visit approach in Accra, Ghana. AB - OBJECTIVE: The purpose of this study was to assess the safety and acceptability of a single-visit approach to cervical cancer prevention combining visual inspection of the cervix with acetic acid wash (VIA) and cryotherapy. STUDY DESIGN: The study was observational. Nine clinicians were trained in VIA and cryotherapy. Over 18 months 3665 women were VIA-tested. If positive and eligible, cryotherapy was offered immediately. Treated women were followed-up at 3 months and 1 year. RESULTS: The test-positive rate was 13.2%. Of those eligible, 70.2% and 21% received immediate or delayed treatment, respectively. No major complications were recorded, and 5.6% presented for a perceived problem post cryotherapy. Among those treated over 90% expressed satisfaction with their experience, and 96% had an indentifiable squamo-columnar junction. Only 2.6% (6/232) were test positive, 1-year posttreatment. CONCLUSION: A single-visit approach using VIA and cryotherapy proved to be safe, acceptable, and feasible in an urban African setting. PMID- 17403439 TI - Predicting gestational diabetes: choosing the optimal early serum marker. AB - OBJECTIVE: Serum markers measured early in pregnancy have been associated with the later diagnosis of gestational diabetes mellitus. To select an optimal early (<20 weeks) marker, we prospectively compared 3 serum markers examined simultaneously in a single cohort. STUDY DESIGN: A nested case-control design was used to evaluate the association of sex hormone-binding globulin, high-sensitive C-reactive protein, and measures of fasting glucose and insulin (homeostasis assessment model) obtained in the late first trimester and early second trimester of pregnancy with the diagnosis of gestational diabetes mellitus. Multivariate modeling and log likelihood ratios were used to identify the optimal biomarker associated with gestational diabetes mellitus. RESULTS: In both first and second trimester samples, sex hormone-binding globulin was lower and high-sensitive C reactive protein higher among women who subsequently developed gestational diabetes mellitus. Similarly an elevated second-trimester homeostasis assessment model was associated with gestational diabetes mellitus. Multivariate analysis suggested that sex hormone-binding globulin measured from nonfasting first trimester sera was the best predictor of gestational diabetes mellitus in our population. CONCLUSION: Among 3 biomarkers examined prospectively, first trimester nonfasting sex hormone-binding globulin appeared to be the optimal marker to predict subsequent gestational diabetes mellitus. PMID- 17403440 TI - Oral contraceptive discontinuation: do side effects matter? AB - OBJECTIVE: The purpose of this study was to assess self-reported side effects in women after they received the oral contraceptive (OC) and to compare discontinuation rates, according to presence or absence of side effects. STUDY DESIGN: The study comprised 1716 women aged <25 years who initiated the OC at 3 publicly funded family planning clinics and completed structured interviews after 3 and 6 months. RESULTS: Nearly 60% of subjects discontinued the OC by 6 months. Most subjects reported no changes in headaches, weight, moodiness, and sexual satisfaction during the first 3 months of OC use. Subjects with any complaints, especially women with increased headaches or moodiness, were more likely to discontinue the OC prematurely. Nonetheless, most discontinuation occurred for reasons that were unrelated to side effects. CONCLUSION: Side effects are absent or mild among most OC users, but women with complaints are more likely to discontinue. Side effects are less important reasons for discontinuation than widely believed. PMID- 17403442 TI - The case of the vanishing tumor. PMID- 17403443 TI - Biomarkers and classifications. PMID- 17403447 TI - Relationship between human herpesvirus 8 loads and disease stage in classic Kaposi sarcoma patients. AB - Human herpesvirus 8 (HHV-8), also known as Kaposi sarcoma (KS)-associated herpesvirus, is causally implicated in all forms of KS, including the classic form. Our objective was to investigate the relationship between HHV-8 load in peripheral blood mononuclear cells (PBMCs) and the stage of the disease in classic KS (CKS) patients. HHV-8 loads were measured in 41 PBMC samples from CKS patients with different Krigel-based classification stages using a quantitative real-time polymerase chain reaction assay. Low HHV-8 DNA loads reaching a maximum of 75.5 copies/10(5) cells were detected in 73.2% of the patients. HHV-8 loads in patients with stages I and II were similarly distributed. An increased detection rate of HHV-8 DNA, although not statistically significant, was evident in patients diagnosed with CKS stages III and IV. We conclude that the measurements of HHV-8 load in PBMCs provide a limited correlation with the clinical stage of KS. PMID- 17403448 TI - Antimicrobial susceptibility of extended-spectrum beta-lactamase producers and multidrug-resistant Acinetobacter baumannii throughout the United States and comparative in vitro activity of tigecycline, a new glycylcycline antimicrobial. AB - As part of the Tigecycline Evaluation and Surveillance Trial, isolates of Escherichia coli, Klebsiella pneumoniae, and Acinetobacter baumannii were collected in the United States between January 2004 and January 2006. Determinations of antimicrobial susceptibility and extended-spectrum beta lactamase (ESBL) production were carried out according to the Clinical and Laboratory Standards Institute guidelines. A high percentage of ESBL-producing K. pneumoniae (>or=19.0%) was detected in New Jersey, Massachusetts, New York, and Missouri, and for E. coli, in the District of Columbia (9.5%). Against ESBL producing isolates, the lowest MIC(90)s were for tigecycline (0.5-2 microg/mL) and imipenem (0.5-8 microg/mL). Overall, 282 (27.5%) A. baumannii isolates were resistant to >or=3 antimicrobial classes. The most common phenotype (33.0%) was resistance to cefepime, ceftazidime, ceftriaxone, levofloxacin, and piperacillin tazobactam. Against multidrug-resistant A. baumannii, tigecycline and minocycline were the most active agents (MIC(90), 2 and 8 microg/mL, respectively). PMID- 17403451 TI - Surgery for atrial fibrillation: a worldwide review. AB - Maze procedure is highly effective in converting atrial fibrillation (AF) back to sinus rhythm and significantly prevents thromboembolism postoperatively. However, the procedure has not been widely performed by many surgeons, because of the technical demand and potential risk of complications of the procedure. During the past several years, the surgical strategy for AF has evolved dramatically and significantly. The evolution can be classified into two strategies: simplification of the lesion set and development of ablation devices. Isolation of the pulmonary veins with or without left atrial incisions has been shown to cure AF in selected patients. During the past decade, a number of ablation devices have been developed to replace the cut-and-sew lesions of the maze procedure and lessen the invasiveness of the procedure. The challenge in AF surgery is in the development and establishment of an off-pump thoracoscopic procedure in the patients with isolated AF. In addition to the development of ablation devices, intraoperative electrophysiological assessment of the triggers and substrates of AF for a step-by-step tailored approach and verification of conduction block over the ablation line should be established to accomplish a high success rate for AF. PMID- 17403449 TI - The evolution of the conservative approach to breast cancer. AB - The profound revolution that surgical treatment of breast cancer has undergone during the past 30 years has led to the progressive reduction of the extent of surgery, with less mutilation. As a consequence, quality of life has improved and women are now more motivated to follow screening programs for early diagnosis of the disease. Since conservative surgery is as effective as radical surgery, research is now focused on reducing radiotherapy. Overall, survival after breast cancer is not affected by reducing the extent of surgery, which, together with less invasive diagnostic procedures, has a good effect on patients' quality of life. For this reason in our Institute we are now evaluating the feasibility of a reduction of the radiation field and the sensibility and sensitivity of new diagnostic approaches for axillary staging. PMID- 17403452 TI - Recent insights into the pathophysiology of atrial fibrillation. AB - Although the problem of atrial fibrillation is now widely appreciated, the fundamental mechanisms that lead to arrhythmia onset and persistence have been difficult to elucidate. As a result, available pharmacologic therapies have focused more on modifying ion channel activity than on the underlying mechanisms. Recent studies suggest an important role for alterations in autonomic regulation, neurohormonal activation, and a systemic inflammatory state in the genesis and persistence of atrial fibrillation. The relative contributions of these distinct pathways to atrial fibrillation likely vary from patient to patient, and within a patient, as a function of age. Tailored therapies, together with patient-specific ablative interventions, may increase the success with which atrial fibrillation is treated and minimize the occurrence of life-threatening thromboembolic complications. PMID- 17403453 TI - An overview of energy sources in clinical use for the ablation of atrial fibrillation. AB - Recent years have seen many developments in the field of alternative energy sources for arrhythmia surgery. The impetus behind these advances is to replace the traditional, "cut-and-sew" Cox maze III procedure with lesion sets that are simpler, shorter, and safer but just as effective. There is demand for technology to make continuous, linear, transmural ablations reliably with a versatile energy source via an epicardial approach. This would make minimally invasive endoscopic surgical ablation of atrial fibrillation (AF) without cardiopulmonary bypass and with a closed chest feasible. These advances would shorten cardio-pulmonary bypass and improve outcomes in patients having surgical ablation and concomitant cardiac surgery. This review summarizes the technology behind alternative energy sources used to treat AF. Alternative energy sources include hypothermic sources (cryoablation) and hyperthermic sources (radiofrequency, microwave, laser, ultrasound). For each source, the biophysical background, mode of tissue injury, factors affecting lesion size, and advantages and complications are discussed. PMID- 17403454 TI - Ablation of atrial fibrillation with concomitant cardiac surgery. AB - Atrial fibrillation is present in approximately 35% of patients presenting for mitral valve surgery and in 1 to 6% of adult patients undergoing other forms of cardiac surgery. If left untreated, atrial fibrillation is associated with increased morbidity, and, in some subgroups, increased mortality. Therefore, concomitant management of the arrhythmia is indicated in most cardiac surgery patients with preexisting atrial fibrillation. Although the cut-and-sew Cox-maze III procedure is extremely effective, it has been supplanted by newer operations that rely on alternate energy sources to create lines of conduction block. Early and mid-term results are good with a variety of technologies. Choice of lesion set remains a matter of debate, but results of ablation appear to be enhanced by a biatrial lesion set. Targeted areas for improvement in concomitant ablation include acceptance of uniform standards for reporting results, development of improved technology for ablation and intraoperative assessment, and creation of instrumentation that facilitates minimally invasive approaches. PMID- 17403455 TI - Minimally invasive surgery for atrial fibrillation. AB - Although the surgical treatment of atrial fibrillation has become technically simpler and faster with the advent of new ablative technologies, the ultimate goal of treating large numbers of patients for lone AF remains elusive. The ideal procedure would be performed epicardially, on the beating heart, with minimal risk to and impact on the patient, a rapid recovery and high cure rates. As minimal access techniques improve along with device designs and our understanding of pathophysiology, the field continues to move toward this objective. This article provides a brief overview of the progress made, questions yet unanswered and possible future directions. PMID- 17403456 TI - Surgical treatment of atrial fibrillation: a look into the future. AB - The surgical treatment of atrial fibrillation began in 1987, when Dr. James Cox introduced the maze procedure. This operation proved to be extremely effective in curing atrial fibrillation and preventing its most dreaded complication, stroke. However, many surgeons found the operation to be too difficult and invasive. Over the last 5 to 10 years, various groups have tried to develop less invasive approaches using a number of different energy sources to create linear lines of ablation to replace the surgical incisions. This has led to a plethora of new operations for this arrhythmia. There is significant confusion in the literature at the present time as to what is the best lesion pattern and what is the best energy source. It is our feeling that a great deal of this confusion is due to our lack of understanding of the mechanisms of atrial fibrillation and the effect of ablation technology on atrial hemodynamics and electrophysiology. Future progress will require a better understanding of this arrhythmia and continued research into the safety and efficacy of ablation devices. PMID- 17403458 TI - Reversing esophageal discontinuity. AB - Catastrophic esophageal or gastric disruption drive the decision to "disconnect" the esophagus to prevent ongoing mediastinal soilage. The operations used to establish esophageal discontinuity are not standardized and vary widely, the surgeon often focusing on saving the patient's life, not on how alimentary continuity will ultimately be restored. Patients who survive the initial disastrous infectious complications are typically desperate to have further surgery to allow them to eat again. Relatively little is written about the decisions involved in reversing esophageal discontinuity--the timing of the operation, preoperative assessment and preparation, planning and conduct of the operation, and outcome. The nuances of reestablishing alimentary continuity in this disparate patient population are the focus of this article. PMID- 17403459 TI - The supercharged microvascular jejunal interposition. AB - In a distinct subset of patients, traditional gastric conduits are not available as esophageal replacements, due to either intrinsic disease, tumor involvement, or previous surgery. Esophageal reconstruction in this group can prove daunting. Colonic interposition can be considered as an alternative conduit, but the "supercharged" jejunum (SPJ) may also serve as another option for total esophageal reconstruction. With an experienced hospital team familiar with esophageal surgery and microvascular reconstruction, morbidity is acceptable and long-term functional benefits may exist with this conduit. This article will review the indications and potential problems with the SPJ and will try to impart some of the important technical points that have been identified to minimize the risk of this complex but useful esophageal reconstruction. PMID- 17403460 TI - Management of the cervical esophagogastric anastomotic stricture. AB - Esophagogastric anastomotic stricture following esophagectomy with a gastric esophageal substitute can be a vexing problem for the patient and treating physician. We describe the clinical practice at a single center with extensive experience in esophageal surgery for management of this complication. PMID- 17403461 TI - Limited surgical resection and jejunal interposition for early adenocarcinoma of the distal esophagus. AB - The need for radical resection and extensive lymphadenectomy for early adenocarcinoma of the distal esophagus has recently been challenged. Limited surgical resection and endoscopic mucosal ablation techniques are increasingly proposed and used as less invasive alternatives. Available data indicate that a limited resection of the distal esophagus and esophagogastric junction with jejunal interposition is associated with less morbidity and mortality, provides similar oncologic results, and offers a better quality of life as compared with radical esophagectomy. In contrast, endoscopic ablation and mucosectomy techniques are still plagued by high tumor recurrence rates, particularly in patients with incomplete removal of the underlying Barrett's mucosa, multicentric tumors, or tumors invading into the submucosa. Attention to technical details of limited resection and jejunal interposition is, however, required to avoid complications, poor functional results, and the need for reintervention. PMID- 17403462 TI - Complications of esophageal resection and reconstruction. AB - Esophagectomy is an acceptable treatment option for esophageal cancer and various end-stage benign esophageal conditions. However, it still has a significantly high morbidity and mortality. In this review, the most common complications are analyzed using evidenced based data and when applicable, special consideration to operative technique is reviewed. PMID- 17403463 TI - The effect of Medicaid family planning expansions on unplanned births. AB - BACKGROUND: Medicaid covers nearly 50% of all family planning services nationally. Between 1994 and 2001, 11 states implemented demonstration programs that expand coverage of family planning beyond the federally mandated minimum coverage levels. METHODS: We estimate the effect of income- and postpartum-based eligibility expansions on birth rates using states that did not expand coverage as a control for states that did expand coverage. Our data span 1991-2001 and include all 50 states. We also estimate net expansion costs from societal and state perspectives for 5 expansions that published incremental expansion costs. RESULTS: We find that Medicaid eligibility expansions lowered average annual birth rates in all states. Birth rates were reduced on average by 1.95 points in income-based expansions and by 0.87 points in postpartum-based expansions. The cost offset of maternal and child health expenditures of the expansions exceed program costs in all states but California. This result is likely because the objectives and scope of the California program goes beyond just unplanned births, which makes the program cost higher relative to the reduction in births. CONCLUSIONS: Both income- and postpartum-based family planning expansions either yield financial benefits or, at the very least, are cost neutral from the perspective of state governments. Income-based expansions are significantly more effective because eligibility is not limited to only postpartum women. The experience of these early family planning expansions should be a guide for other states considering family planning benefit expansions. From the national perspective, 4 out of 5 programs were cost neutral, although California had significantly higher costs. From the state's perspective, all of the expansions were either budget neutral or yielded a net cost savings. PMID- 17403464 TI - Trends in rates of hospitalization with a diagnosis of substance abuse among reproductive-age women, 1998 to 2003. AB - OBJECTIVE: To describe trends in hospitalizations with a diagnosis of substance abuse among reproductive-age women from 1998-2003. METHODS: Data were obtained from the Healthcare Cost and Utilization Project Nationwide Inpatient Sample. Hospitalizations with a diagnosis of substance abuse were categorized into subgroups by age, primary expected payer, substance-specific diagnoses, concomitance, and hospital location. Trends in hospitalization rates per 100,000 women aged 15-44 were tested using a weighted least-squares method. RESULTS: From 1998-2003, there was no change in the overall rate of hospitalization with a diagnosis of substance abuse among women aged 15-44. Alcohol abuse was the most common substance-specific diagnosis. The rate of hospitalization with a diagnosis of cocaine abuse decreased 22%; for a diagnosis of cannabis abuse, the rate increased 35%. The rate of hospitalization with a diagnosis of amphetamine abuse doubled from 1998-2003. Among women aged 15-24, the rate of hospitalization with a diagnosis of substance abuse increased 23%. CONCLUSION: Although we did not observe a change in the overall rate of substance-abuse hospitalization among reproductive-age women, there were dramatic changes in the rate of substance specific diagnoses. These data may be used to quantify emerging trends in substance abuse and promote the use of hospital-based interventions. PMID- 17403465 TI - Increasing breastfeeding rates: do changing demographics explain them? AB - OBJECTIVES: This study examines whether increases in breastfeeding rates between 1991 and 2002 can be attributed to changes in the demographic characteristics of births. The demographic variables investigated include maternal age, maternal education, race/ethnicity, geographic location of birth, and parity. METHODS: This study decomposes breastfeeding trends using 1991 through 2002 data from the Ross Laboratories Mothers Survey and birth certificates. RESULTS: Changing birth compositions by maternal age and education explain 9.8% and 11.5% of the increase in breastfeeding initiation rates, respectively. Changing birth compositions by maternal age and education explain 10.2% and 9.0% of increasing breastfeeding rates 6 months after birth, respectively. If changes in the composition of births by race/ethnicity had not occurred, breastfeeding rates would have increased more. Changes in the composition of births by geographic location of birth and parity do not explain any of the increase in breastfeeding rates. CONCLUSIONS: Overall, changes in the demographic characteristics of births explain up to approximately 20% of the increasing trends in initial breastfeeding rates and breastfeeding rates 6 months after birth. Although this is a significant amount, it is important to continue investigating which additional factors explain recent breastfeeding increases such as changes in laws and policies, health promotion, the Special Supplemental Program for Women, Infants, and Children, employer support, technological innovation, and attitudes toward breastfeeding. PMID- 17403466 TI - Physician knowledge levels and barriers to coronary risk prevention in women: survey results from the Women and Heart Disease Physician Education Initiative. AB - BACKGROUND: Few studies have examined whether physician knowledge, attitudes, or practice patterns might contribute to gender disparities in the primary prevention of coronary heart disease (CHD), including among physicians caring for the largest number of reproductive-age women, obstetricians and gynecologists (OB/GYNs). We sought to identify barriers affecting the provision of recommended coronary risk factor therapies in women. METHODS: We surveyed internists and OB/GYNs who attended Grand Rounds presentations developed for the New York State Women and Heart Disease Physician Education Initiative. This program was designed to improve screening and management of coronary risk factors in women. Attendees were asked to complete a 7-minute questionnaire. RESULTS: The mean age of the 529 respondents was 40.3 years (standard deviation = 12.3), 75.1% were internists (n=378), and 42.7% (n=226) were women. Physicians correctly responded to 71.5% of the 13 questions assessing knowledge of coronary risk prevention (range, 4-13). Almost one third of internists and half of the OB/GYNs did not know that tobacco use was the leading cause of myocardial infarction in young women. For patients who smoked tobacco, only two thirds of internists and 55.4% of OB/GYNs reported suggesting a quit date (p=.007). After controlling for covariates, physicians who did not perceive time as a barrier were more likely to discuss smoking cessation (odds ratio=1.7 [1.1-2.7]). CONCLUSIONS: Among the internists and OB/GYNs surveyed, time was perceived as a barrier to implementing risk prevention. These physicians also underestimated the impact of tobacco use as a risk factor for CHD in young women. To lessen gender disparities in CHD prevention, both specialties need time-efficient educational programs that reflect specialty differences. PMID- 17403467 TI - Epidemiology and consequences of women's revictimization. AB - This study uses Kraemer's approach for nonrandom comorbidity to identify the parameters of revictimization among women, using a diverse, population-based sample. Participants (n = 11,056) are from the California Women's Health Survey. Women were asked about childhood and adult violence and current symptoms of posttraumatic stress disorder (PTSD), depression, and anxiety. Logistic regressions adjusted for age, ethnicity, education, and poverty indicate that women who experienced childhood physical or sexual abuse were 5.8 (95% confidence interval, 5.2-6.4) times more likely to experience adult physical or sexual victimization. Revictimization affected 12% of women, and these women were substantially more likely to report current symptoms of anxiety, depression, and PTSD than women exposed to violence only in childhood or only as an adult. Revictimization is a methodologically distinct concept and is a potent risk factor for adult mental health problems. Prevention should target women exposed to both physical and sexual assault. PMID- 17403468 TI - Obstetricians, health attorneys, and court-ordered cesarean sections. AB - PURPOSE: Despite court rulings suggesting that court-ordered cesarean sections should rarely be undertaken, they are performed. Our objective was to determine those characteristics of providers and patients that make their use more likely. METHODS: A convenience sample of obstetricians attending the annual meeting of the American College of Obstetricians and Gynecologists (n=229) and lawyers attending the annual meeting of the American Health Lawyers Association (n=126) read a vignette describing a woman who refused a cesarean section after being told that her fetus would die unless she received the operation. Several different scenarios were then described (e.g., the mother refused on religious grounds). For each scenario, participants were asked to rate the likelihood that they would support a court-ordered cesarean section. Participants then described their own characteristics. RESULTS: Respondents were more likely to support a court order if the woman was described as a being "high on drugs" (p<.001). Respondents were significantly less likely to use a court order (p<.01) if the woman was described as a lawyer, the child had Down syndrome, the husband agreed with her decision, the mother refused on religious grounds, or the operation was associated with a 10-fold increased risk to the mother's life. People who described themselves as more religious, Republican, or "pro-life" were significantly more likely to utilize court orders in several scenarios. In an ordinal regression model, the degree of pro-life was the only variable consistently associated with obtaining a court order for the healthy mother healthy child. CONCLUSIONS: The perceived likelihood of performing a court ordered cesarean section varies with characteristics of the patient and the provider. PMID- 17403469 TI - Recommendations for the use of everolimus (Certican) in heart transplantation: results from the second German-Austrian Certican Consensus Conference. AB - Everolimus (Certican; Novartis Pharma AG, Basel, Switzerland) represents the latest generation of proliferation signal inhibitors (PSIs). Everolimus is indicated for use as an immunosuppressive drug in renal and heart transplantation. This report reflects the recommendations of the second German Austrian Certican Consensus Conference, held in January 2006, for the clinical use of everolimus. PMID- 17403470 TI - Pre-existing pulmonary hypertension in patients with end-stage heart failure: impact on clinical outcome and hemodynamic follow-up after orthotopic heart transplantation. AB - BACKGROUND: The purpose of this investigation was to determine the influence of pre-existing pulmonary hypertension (PHT) on outcome and to assess the pulmonary hemodynamic changes after heart transplantation (HT). METHODS: A total of 410 patients were studied before and after (1 month and 1 year) HT: Group 1 (n = 266) had no PHT (PVR < 3 Wood units [WU], TPG < 10 mm Hg); Group 2 (n = 112) had mild moderate PHT (PVR 3 to 6 WU, TPG 10 to 20 mm Hg); and Group 3 (n = 32) had severe PHT (PVR > 6 WU, TPG > 20 mm Hg). RESULTS: Mean (+/- SD) follow-up was 5.2 +/- 4.1 years, mean recipient age was 57 +/- 11, and mean donor age was 30 +/- 12 years. Baseline characteristics were similar in all groups, except donor/recipient weight ratio, which was higher in patients with PHT (p = 0.002). There was a significant (p < 0.0001) decrease in mean TPG to 11.0 within the first month and to 9.5 mm Hg after the first year. Decreases in PVR to 2.2 and 2.0 WU at 1 month and 1 year, respectively (p < 0.0001 for both) were also found. Reversibility (after vasodilation) of PHT was obtained in 85% of patients in Group 2 and in 84% in Group 3. Patients' PHT did not show a significant difference in 30-day mortality (p = 0.9) and long-term survival (p = 0.8). Patients with residual post-transplant PHT (PVR > or = 3 WU) had reduced long term survival (p = 0.03). Multivariate analysis showed no evidence that elevated PVR was associated with death. CONCLUSIONS: Pre-existing elevated PVR that responds to vasodilator challenge does not have a negative influence on short- and long-term survival after HT. We found that residual post-transplant PHT is associated with decreased long-term survival. PMID- 17403471 TI - Effect of reversible pulmonary hypertension on outcomes after heart transplantation. AB - BACKGROUND: Conflicting data exist regarding the impact of reversible pulmonary hypertension (PHTN) on post-transplant (Tx) outcomes. In this study we sought to determine the influence of reversible PHTN on outcomes after Tx. METHODS: We retrospectively reviewed the records of adult patients who underwent heart Tx from 1993 to 2002. Patients were grouped depending on their measured pulmonary vascular resistance (PVR). Group 1 patients had a pre-Tx pulmonary vascular resistance (PVR) of < 3 Wood units (WU). Patients with reversible PHTN, defined as pre-Tx PVR > or = 3 WU and reversing to < 3 WU either with sub-lingual or intravenous vasodilatory agents, were divided into two groups based on their PVR before the reversibility test (PVR: Group 2, 3 to 4.5 WU; Group 3, > 4.5 WU). RESULTS: Records for 222 adult heart recipients were reviewed (Group 1, n = 171; Group 2, n = 35; Group 3, n = 16). Baseline clinical characteristics (age, gender, heart failure etiology, history of diabetes, ischemic time, donor age and gender) were similar in the three groups and the average follow-up was 58 months. One-month and 1-year mortality (Groups 1, 2 and 3: 2%, 0% and 13%; and 8%, 0% and 13%, respectively) did not differ significantly between groups. Actuarial mortality was assessed using Cox regression analysis, adjusted for age and gender, and no increased risk of death was demonstrated for patients with reversible PHTN (for Group 2: multivariate hazard ratio [HR] 0.45, 95% confidence interval [CI] 0.17 to 1.32, p = 0.15; for Group 3: HR 0.98, CI 0.34 to 2.84, p = 0.97). No differences were observed between the three groups for various post-Tx events, such as hospital stay, ICU stay, extubation time, transfusions, acute allograft dysfunction, acute hepatic dysfunction, acute and chronic renal dysfunction, infections, neurologic complications, gastrointestinal complications and coronary allograft vasculopathy. CONCLUSIONS: Reversible pulmonary hypertension is associated with similarly good post-transplant survival outcomes and morbidity, regardless of severity. PMID- 17403472 TI - Interplay between systemic inflammation and markers of insulin resistance in cardiovascular prognosis after heart transplantation. AB - BACKGROUND: Metabolic and immuno-inflammatory risk factors contribute to cardiac allograft vasculopathy (CAV) pathogenesis. Although systemic inflammation, as detected by C-reactive protein (CRP), predicts CAV development, the relationship between CRP and markers of metabolic abnormalities remains unexplored. METHODS: CRP and the entire metabolic panel were evaluated in 98 consecutive heart transplant recipients at the time of annual coronary angiography, 5.8 years after transplant (range, 1-12 years). A ratio of triglycerides (TG) to high-density lipoproteins (HDL) of 3.0 or more was considered a marker of insulin resistance. CAV prevalence was defined by angiography, and subsequent prognosis was evaluated as incidence of major cardiac adverse events. RESULTS: CRP was higher in the 34 patients with angiographic CAV than in those without CAV (1.10 +/- 0.20 vs 0.50 +/- 0.05 mg/dl, p < 0.001). Patients with insulin resistance had higher CRP concentrations (p = 0.023) and higher CAV prevalence (p = 0.005). High CRP and a TG/HDL of 3.0 or more were independently associated with an increased likelihood of CAV (odds ratio, > or = 3.9; p = 0.02) and predicted an increased risk of major cardiac adverse events. The combination of high CRP and a TG/HDL of 3.0 or more identified a subgroup of patients having a 4-fold increased risk for CAV and a 3-fold increased risk for major cardiac adverse events compared with patients with low CRP and normal values for metabolic indicators. CONCLUSIONS: Both CRP and insulin resistance, as estimated by TG/HDL, appear to be strong, synergic risk factors for CAV and for major cardiac adverse events. These findings support the hypothesis that in heart transplant recipients, systemic inflammation may be an important mediator of graft vascular injury associated with metabolic syndrome. PMID- 17403473 TI - Early institution of extracorporeal membrane oxygenation for primary graft dysfunction after lung transplantation improves outcome. AB - BACKGROUND: Primary graft dysfunction (PGD) after lung transplantation (LTx) carries a significant mortality and clinical management is controversial. Extracorporeal membrane oxygenation (ECMO) has been used infrequently for recovery from acute lung injury (ALI) in this setting. We reviewed our experience with ECMO after primary LTx. METHODS: The present study is a retrospective analysis of all LTx patients between 1991 and 2004. Twenty-two patients sustained severe PGD with subsequent placement on ECMO. We analyzed indications and 30-day, 1-year and 3-year mortality. Complications and incidence of multiple-organ failure (MOF) were determined. Critical appraisal of the evidence available to date was performed. RESULTS: A total of 297 LTxs were performed during the study period, with 97.5%, 88.6% and 73.8% survival at 30 days, 1 year and 3 years, respectively. Twenty-two patients (7.9%) had severe allograft dysfunction leading to ECMO support. Twelve patients received single-lung (SLTx), 8 double-lung (BLTx), 1 single-lung/kidney (SLKTx) and 1 heart/lung (HLTx) transplantation. Thirty-day, 1-year and 3-year survival of LTx recipients with ECMO support post operatively were 74.6%, 54% and 36%, respectively. MOF was the predominant cause of death (58.3%) in patients on ECMO support for PGD. CONCLUSIONS: Our data suggest that, in addition to prolonged ventilation and pharmacologic support, ECMO should be considered as a bridge to recovery from PGD in lung transplantation. Early institution of ECMO may lead to diminished mortality in the setting of ALI despite the high incidence of MOF. Late institution of ECMO was associated with 100% mortality in this investigation. PMID- 17403474 TI - High incidence of venous thromboembolic events in lung transplant recipients. AB - BACKGROUND: Previous studies have reported a 12% incidence of venous thromboembolic events (VTEs) in lung transplant recipients (LTRs). Characterization of risk factors for VTEs in LTRs is lacking. We identified the incidence and risk factors associated with post-transplant VTEs. METHODS: A retrospective review of 153 LTRs from 1994 to 2006 was performed. Patients were categorized by age, race, gender, weight, underlying diagnosis, procedure, ischemic time, length of stay (LOS), cardiopulmonary bypass (CPB), location and number of VTEs, mobility, immunosuppression, renal, hepatic, hematologic and coagulation profiles and nutritional status. RESULTS: A single VTE occurred in 29% of LTRs within the study period. Fifty-eight percent had multiple VTEs and 7% had a radiologically confirmed pulmonary embolism. Median time from transplant to first VTE was 69 days. Sixty percent of VTEs occurred within 1 year, 20% of which occurred within the first month, 19% between 2 and 5 years, and 13% at beyond 5 years post-transplant. Seventy-six percent of VTEs occurred during hospitalization, 19% during outpatient status. Forty-eight percent were of the upper extremity and 47% were of the lower extremity. Sixty-one percent of LTRs were taking cyclosporine and 39% tacrolimus. VTE and non-VTE groups were similar in age, weight, body mass index (BMI), ischemic time, procedure or underlying diagnosis precipitating the need for transplant. Univariate analysis revealed LOS and CPB as significant predictors of a single VTE (p = 0.036, hazard ratio [HR] 1.006 and p = 0.045, HR 1.91, respectively). Multivariate analysis revealed only CPB as a significant predictor (p = 0.047, HR 1.929). CONCLUSIONS: Analysis of a cohort of LTRs for a median period of 1.5 years revealed a VTE incidence much higher than previously reported, especially within the first month after transplantation. PMID- 17403475 TI - Levitronix ventricular assist devices as a bridge to recovery after profound biventricular heart failure associated with pulmonary aspergillosis. AB - A patient with multisystem organ failure and refractory cardiopulmonary shock stemming from Aspergillus pneumonia was treated with 2 Levitronix ventricular assist devices as a bridge-to-recovery. After ventricular assist device placement, the patient recovered myocardial function. The ventricular assist devices were removed on post-implant Day 7, and the patient made a full long-term recovery. Ventricular assist devices should be strongly considered, as bridges to recovery, to support patients with acute myocardial dysfunction associated with sepsis while the underlying infection is treated. PMID- 17403476 TI - Scedosporium apiospermum (Pseudoallescheria boydii) infection in lung transplant recipients. AB - BACKGROUND: Scedosporium apiospermum, the asexual counterpart of the teleomorph Pseudallescheria boydii, is increasingly recognized as an important opportunistic pathogen in transplant recipients. Infection is associated with a high rate of dissemination and poor outcome overall. METHODS: A retrospective analysis of The Cleveland Clinic lung transplant database identified 5 patients with S. apiospermum isolated from respiratory tract specimens. Demographic data and lung transplant outcomes were obtained by review of medical records. RESULTS: S. apiospermum was isolated from respiratory culture in 5 lung transplant recipients. Disseminated disease developed in 3 patients, whereas 2 appeared only to be colonized. CONCLUSIONS: Our experience and review of the literature highlights the importance of early diagnosis and differentiation from Aspergillus, since Scedosporium is inherently resistant to amphotericin B. Effective therapeutic approaches being explored include combinations of anti fungals, because even the newer triazoles have a 50% response rate in clinical studies. Surgical debridement and immune recovery are associated with improved prognosis, favoring the use of agents that expedite immune reconstitution in these patients. Close monitoring of clinical improvement and frequent reevaluation of treatment is essential. PMID- 17403477 TI - Intravenous epoprostenol in inoperable chronic thromboembolic pulmonary hypertension. AB - BACKGROUND: Pulmonary thromboendarterectomy may be a cure for patients with chronic thromboembolic pulmonary hypertension (CTEPH) with proximal obstruction. Conversely, short-term prognosis is poor for inoperable CTEPH patients with distal obstruction. Vascular remodeling lesions can be found in lungs of patients with CTEPH. Little is known about epoprostenol therapy in inoperable distal CTEPH patients. METHODS: We conducted a retrospective analysis of 27 consecutive patients with inoperable distal CTEPH, who were in New York Heart Association Functional Class (NYHA FC) III (n = 20) or IV (n = 7) and treated with long-term intravenous epoprostenol. RESULTS: Before epoprostenol initiation (baseline), 6 minute walk distance (6MWD) was 265 +/- 117 m, right atrial pressure 13 +/- 5 mm Hg, mean pulmonary artery pressure (mPAP) 56 +/- 9 mm Hg, cardiac index (CI) 1.9 +/- 0.3 liters/min/m2, total pulmonary resistance (TPR) 29.3 +/- 7.0 U/m2 and mixed venous oxygen saturation 52 +/- 8%. After 3 months of epoprostenol (n = 23; mean epoprostenol dose 16 +/- 2 ng/kg/min), NYHA FC improved by one class in 11 of 23 patients, 6MWD increased by 66 m (p < 0.0001), and hemodynamics also improved (mPAP: 51 +/- 8 mm Hg, p = 0.001; CI: 2.3 +/- 0.4 liters/min/m2, p = 0.0003; TPR: 23.0 +/- 5.0 U/m2, p < 0.0001). At last evaluation (n = 18; 20 +/- 8 months), mean dose of epoprostenol was 30 +/- 8 ng/kg/min, NYHA FC was improved in 9 of 18 patients and there was sustained improvement in 6MWD (+46 m, p = 0.03) and hemodynamic parameters. Survival at 1, 2 and 3 years was 73%, 59% and 41%, respectively. CONCLUSIONS: Long-term treatment with intravenous epoprostenol improves exercise capacity and hemodynamic parameters in patients with inoperable distal CTEPH. Epoprostenol may be used as bridge therapy in CTEPH patients awaiting lung transplantation. PMID- 17403478 TI - Long-term follow-up after conversion from intravenous epoprostenol to oral therapy with bosentan or sildenafil in 13 patients with pulmonary arterial hypertension. AB - BACKGROUND: Epoprostenol significantly improves function and survival in patients with pulmonary arterial hypertension (PAH) but is associated with many risks and side effects. Furthermore, effective oral therapy is now available. We report our long-term experience with 13 patients from among 118 treated with epoprostenol who were able to be weaned to oral therapy, including 6 with persistently abnormal hemodynamics (mean pulmonary artery pressure > or = 35 mm Hg). METHODS: Oral therapy with bosentan (n = 11) or sildenafil (n =2) was started before weaning epoprostenol in all but 1 patient. Right heart catheterization was performed when patients reached a dose of 2 ng/kg/min, and epoprostenol was discontinued with hemodynamic monitoring. Functional class and 6-minute walk test were assessed at regular intervals. Repeat right heart catheterization was performed 1 year after discontinuation of epoprostenol. RESULTS: Nine patients remained on oral therapy alone for up to 46 months. Four patients deteriorated in functional class, and 2 of them resumed epoprostenol therapy. Inhaled iloprost was started in another patient. One additional patient died, unrelated to PAH. Twelve patients underwent right heart catheterization at the time of epoprostenol discontinuation. Hemodynamic evaluation 13.2 +/- 0.9 months later showed that the 5 patients with normal or nearly normal hemodynamics at the time of discontinuation of epoprostenol had no deterioration, whereas 4 of the 7 patients with abnormal hemodynamics had worsened. The 6-minute walk test at last follow-up was not significantly changed from maximal distance on epoprostenol (420 +/- 94 vs 412 +/- 95 meters). CONCLUSION: Weaning from epoprostenol to sildenafil or bosentan with sustained clinical improvement is possible, even with persistent pulmonary hypertension; however, patients with persistently abnormal hemodynamics are at risk for hemodynamic and clinical deterioration and require close follow up. PMID- 17403479 TI - Sivelestat reduces reperfusion injury of lungs harvested from endotoxin-primed rats by inhibition of neutrophil-mediated inflammation. AB - BACKGROUND: Although liberalization of donor criteria may be one of the solutions to the current serious lung donor shortage, the use of non-standard donor lungs would increase the risk of post-operative complications. In the present study, we investigated the effect of sivelestat, a neutrophil elastase inhibitor, on reperfusion injury of a donor lung that was harvested from endotoxin-primed animals in a rat lung transplantation model. METHODS: Donor rats received an intraperitoneal injection of Escherichia coli endotoxin (5 mg/kg) 2 hours before lung harvesting. The donor lungs were flushed with an organ preservation solution with or without sivelestat (300 microg/ml), and the left lung was immediately transplanted to the recipient by the cuff technique. RESULTS: Endotoxin priming did not cause significant lung injury before harvesting. Although these lungs looked normal macroscopically, they were found to contain numerous neutrophils in the alveolar capillaries, even after lung flushing. There was no significant difference in the neutrophil count between the lungs flushed with and without sivelestat. The endotoxin-primed donor lung without sivelestat treatment became edematous immediately after reperfusion. In addition, the recipient's native right lungs were also pathologic. Treatment with sivelestat significantly reduced injury in both the donor and the recipient's native lungs. Treatment with sivelestat also inhibited the increase in tumor necrosis factor-alpha and cytokine-induced neutrophil chemoattractant-1 levels in the recipient circulation after reperfusion. CONCLUSIONS: We conclude that sivelestat could reduce lung injury after transplantation by inhibiting the deleterious burst of inflammatory reactions that are initiated by reperfusion of the lungs from endotoxin-primed rats. PMID- 17403480 TI - Use of diffusion tensor imaging to predict myocardial viability after warm global ischemia: possible avenue for use of non-beating donor hearts. AB - BACKGROUND: The assessment of myocardial viability after global warm ischemia (WI) but before reperfusion is challenging. We hypothesized that fractional anisotropy (FA), a magnetic resonance imaging (MRI) parameter of water diffusion that characterizes cellular integrity within tissues, provides a rapid and useful method for evaluating the viability of hearts after WI. METHODS: Dog hearts were exposed to 60 minutes of WI after exanguination, explanted and preserved in a cold, non-beating state for 6 hours, using continuous perfusion (CP) or static cold storage (CS). Toward the end of preservation, a global FA assessment, acquired using MRI, was compared with analyses obtained from myocardial biopsies that included adenosine triphosphate (ATP), endothelin-1 (ET-1) and caspase-3 levels, light microscopy and tetrazolium staining. Functional recovery was analyzed after restoration of blood flow on a non-working Langendorff preparation. RESULTS: FA measured at the end of CP showed strong correlations with all parameters of functional recovery (developed pressure, R = 0.60; dP/dt, R = 0.96; -dP/dt, R = 0.96). Although FA also correlated with tissue levels of ATP, ET-1 and caspase-3 (R = 0.77, -0.84, -0.64), recovery of myocardial function did not correlate with these markers or any other conventional analyses of myocardial injury (troponin I, changes on light microscopy or tetrazolium staining). CONCLUSIONS: FA, an MRI-based parameter that indicates cellular integrity, was found to reflect better myocardial ATP stores, less induction of ET-1 and caspase-3 and improved functional recovery of hearts after global WI. As a clinically applicable tool capable of rapidly differentiating reversible from lethal injury, diffusion tensor imaging may prove useful in the eventual adoption of non-beating donor hearts for transplantation. PMID- 17403481 TI - Mechanical circulatory support as a bridge to heart transplantation: what remains? Long-term emotional sequelae in patients and spouses. AB - BACKGROUND: Implantation of a ventricular assist device (VAD) reduces short-term mortality and morbidity and provides patients with reasonable quality of life even though it may also be a long-lasting emotional burden. This study was conducted to analyze the long-time emotional consequences of VAD implantation, followed by heart transplantation in patients and spouses. METHODS: This cross sectional study used the Impact of Event Scale-Revised (IES-R) Version, recording avoidance, intrusion, and hyperarousal, to investigate symptoms of post-traumatic stress disorder (PTSD), and VAD-related fears and concerns. The study cohort comprised 38 patients (36 men, 2 women) and 27 spouses (26 women, 1 man), 6 to 135 months post-operatively. RESULTS: Seven (26%) of the 27 spouses but none of the patients met the criteria for PTSD. Patients who were operated at the early stage of our VAD program (82.0 +/- 31.4 months between implantation and evaluation) were significantly more likely to have a spouse with PTSD syndromes than those whose operation took place later on (42.1 +/- 31.1 months, p = 0.007). Patients with higher avoidance scores and a higher level of hyperarousal were significantly more often affiliated with a PTSD spouse than those with lower avoidance (p = 0.008) and hyperarousal scores (p = 0.001). Spouses displayed significantly higher scores in all IES-R dimensions, and they worried more about device-related problems (malfunctioning, pain, infection, and stroke) than the patients themselves. The noise of the device system was not a crucial issue. CONCLUSION: Our study found that implantation of a VAD, followed by transplantation, does not lead to PTSD in patients but it does in their spouses in the long run. Their emotional well being deserves much closer attention. PMID- 17403482 TI - Lung transplantation in the Fischer 344-->Wistar Kyoto rat strain combination is not suitable to study bronchiolitis obliterans. AB - BACKGROUND: To elucidate the pathogenesis of bronchiolitis obliterans (BO) a reliable animal model is needed. According to the literature, lung transplantation from Fischer 344 (F344) to Wistar Kyoto (WKY) rats is the only model that reliably results in BO without a further stimulus. METHODS: We performed orthotopic left lung transplantation in F344 to WKY rats and in both isogeneic rat strain combinations. Suture and cuff techniques for anastomosis were compared. The time course of rejection and the morphology of the bronchial anastomoses were documented by repeated flat-panel volumetric computed tomography (fpVCT) in the living animal. Graft histopathology was analyzed 3 months post transplant. RESULTS: According to the graft outcome, as revealed by fpVCT, grafts were sub-divided into two groups: In Group 1, infiltrates due to acute rejection occurred early after transplantation and resolved thereafter. Graft histopathology showed minor changes but no BO. In Group 2, acute rejection caused total atelectasis that never resolved. After 3 months, grafts were shrunken and exhibited tissue remodeling with some similarities to BO. No correlation between graft outcome and anastomotic technique was apparent. CONCLUSIONS: Modeling lung transplantation using the F344-to-WKY combination is without clinical relevance because BO does not develop in grafts with life-sustaining function. Consecutive fpVCT is useful to monitor pathologic changes in rat pulmonary grafts. PMID- 17403483 TI - Varicella infection after heart and lung transplantation: a single-center experience. AB - Disseminated varicella-zoster virus infection after organ transplantation in adults is a rare but serious event causing significant morbidity and mortality. We describe our 10-year experience of 13 cases in a single center, including risk factors for infection, lack of protection from pre-existing anti-varicella-zoster virus antibodies, and unusual modes of presentation, including disseminated intravascular coagulation. We also report our preliminary observation of resolution of infection without sequelae in 4 patients with severe disseminated varicella-zoster virus infection who were treated with the combination of intravenous acyclovir and polyspecific intravenous immunoglobulin. PMID- 17403484 TI - Transplant vasculopathy is associated with increased AlloMap gene expression score. AB - The AlloMap gene expression test is used for the non-invasive detection of rejection in heart transplant recipients. We evaluated the impact of transplant vasculopathy on AlloMap gene expression analysis. A total of 69 heart transplant recipients, mean age 53 years, were evaluated at a mean 35 months post transplant. AlloMap score was determined on the same day of the endomyocardial biopsies. Twenty patients had evidence of vasculopathy by coronary angiography (vasculopathy group). These were compared to the remaining 49 patients (control group). The vasculopathy group had a longer mean follow-up duration (48.7 vs 28.8 months, p < 0.01), lower left ventricular ejection fraction (51% vs 60%, p < 0.01) and increased use of sirolimus (40% vs 16%, p = 0.034) compared with controls. Using the logistic regression model and bagging bootstrap approach to adjust for the time factor and potential confounders, the vasculopathy group had a significantly higher AlloMap score than the control group (32.2 +/- 3.9 vs 26.1 +/- 6.5, p < 0.001). There was a correlation of AlloMap score with time after transplantation (r = 0.31, p = 0.01). We found transplant vasculopathy to be associated with increased AlloMap score. PMID- 17403485 TI - Broncholith caused by donor-acquired histoplasmosis in a lung transplant recipient. AB - A broncholith is a calcified lymph node that erodes into and partially or completely obstructs the bronchial lumen. The natural history of broncholiths is poorly understood. They are frequently encountered in residents of areas that are endemic for Histoplasma capsulatum and Mycobacterium tuberculum. We report the first case of a broncholith in which the fungus Histoplasma capsulatum was transferred from a donor to a lung transplant (LTx) recipient. Our report highlights the time course of broncholith development and its successful management. We suspect that broncholithiasis and transmission of Histoplasma capsulatum from a donor to the recipient are under-reported in the LTx literature. We hypothesize that histoplasmosis can be transmitted from the donor to the recipient and the duration in the formation of calcification of the lymph node or the broncholith can be anywhere from 2 to 10 months. PMID- 17403486 TI - Rectal and cutaneous malakoplakia in an orthotopic cardiac transplant recipient. AB - Malakoplakia, from the Greek term meaning "soft plaque," is a rare granulomatous disease first documented by Michaelis and Gutmann in 1902, and is most commonly found in association with systemic diseases such as systemic lupus erythematosus, tuberculosis, diabetes mellitus, sarcoidosis or neoplasm. We present a case report of malakoplakia in a 55-year-old female patient who had undergone an orthotopic cardiac transplant for ischemic cardiomyopathy. She was found to have an irregular rectal mass, after presenting with a complaint of hematochezia and pressure with defecation as well as a "boil" on her right labia. Malakoplakia is an exceedingly rare disease entity, and this is only the third known report of this condition in a patient after cardiac transplantation. We review the current perspectives on the pathophysiology, diagnosis and treatment of this disease. PMID- 17403487 TI - Recipient atrial flutter masquerading as donor flutter in a heart transplant recipient. AB - Atrial flutter is common after orthotopic heart transplantation. We present the case of a patient who presented with atrial flutter on 12-lead electrocardiogram. On invasive electrophysiologic study, the donor heart was found to be in sinus rhythm and the recipient atrium was in flutter, which was dissociated from the donor. PMID- 17403488 TI - Intracardiac giant cells after left ventricular assist device placement. AB - Multinucleated giant cells are seen in the heart in several distinct pathologic conditions, including giant cell myocarditis, sarcoidosis and infectious etiologies. We describe the first report of solitary multinucleated giant cells scattered throughout the heart after placement of a left ventricular assist device (LVAD) and transient Candida glabrata infection. The pathologic findings were noted in the explanted heart of a patient undergoing orthotopic cardiac transplantation 7 months after LVAD surgery. The presence of solitary giant cells without adjacent inflammation was a distinct histologic finding in this case. An awareness of this entity is important to avoid making a diagnosis of a more severe giant cell-related pathologic disease, which may recur in a transplanted heart. PMID- 17403489 TI - Soluble human leukocyte antigen-G: a new strategy for monitoring acute and chronic rejections after heart transplantation. PMID- 17403490 TI - Cultural dynamics in HIV/AIDS prevention research among young people. AB - In September 2005, the National Institute of Nursing Research with the support of several other components of the National Institutes of Health held a workshop, "Cultural Dynamics in HIV Biobehavioral Research." This special issue of the Journal of the Association of Nurses in AIDS Care contains a series of articles developed from that workshop. The articles are derived from those presentations that focused on prevention of infection. They do not contain an agenda or recommendations from the National Institute of Nursing Research or any other component of the National Institutes of Health. Rather, the purpose of this special issue is to share with a broad audience the exciting dialogue that began at the workshop and which the authors hope will continue for some time in the future. The articles represent an interdisciplinary and global perspective. Issues discussed include behavioral theory, intergenerational communication, historical trauma, modernization, research methodology, and the ethics of community clinic trials. PMID- 17403491 TI - Scaffolded interviewing with lesbian, gay, bisexual, transgender, queer, and questioning youth: a developmental approach to HIV education and prevention. AB - The field of education has long recognized that adolescent development and learning are made possible by the structural supports or "scaffolds" that adults create with young people. Although the work of Lev Vygotsky (1978, 1987) has inspired developmentally-supportive approaches to education in the United States and internationally, his work has been largely overlooked in the field of HIV education and prevention. This article introduces an approach to scaffolded interviewing that builds narrative and relational "platforms" for young people's self-development and facilitates health communication, trust and rapport, and HIV awareness. Developed over the course of a 2-year longitudinal ethnographic study with 45 lesbian, gay, bisexual, transgender, queer, and questioning youths, scaffolded interviewing aims to build the relational and narrative foundations for young people's active engagement in HIV education and prevention. In scaffolded interviewing, three kinds of platforms or supportive structures serve to scaffold enhanced health communication and HIV awareness: (a) the interview design (a strategic sequencing of life history and HIV-related questions), (b) the developing relationship between interviewer and study participant, and (c) the young person's own narration of a "real" and developing self. Through their participation in scaffolded interviewing, young people develop their own foundations for HIV awareness and HIV prevention by using the narrative and relational supports of the research or clinical interview and the identity terminologies relevant to their own self-development. PMID- 17403492 TI - Research brief: the need for historically grounded HIV/AIDS prevention research among Native Americans. AB - This is a brief report that summarizes the need for historically grounded HIV prevention research among Native Americans living in the United States. It illustrates the intersection of culture and history, showing that ethnic groups can respond to historical traumatic events for generations, often to the detriment of individual and collective health. PMID- 17403493 TI - Childhood abuse history and risk behaviors among teen parents in a culturally rooted, couple-focused HIV prevention program. AB - Pregnant and parenting adolescents living in inner cities are at risk for acquiring HIV through unprotected sexual activity. In addition to individual risk behaviors, a lack of socioeconomic and other environmental resources create risk environments that make certain communities vulnerable to both adolescent pregnancy and HIV/AIDS. Research indicates that adolescent parents, many who have histories of childhood trauma, may use their experience of young parenthood and the concomitant feelings of parental protectiveness as a source of renewed hope for their future. The purpose of this report is to explore the relationship between history of childhood abuse and high risk behaviors in adolescent Latino mothers and fathers enrolled in a randomized clinical trial of a culturally rooted, couple-focused HIV prevention program. In addition, this report describes the HIV prevention program that was designed specifically for young Latino parents wherein maternal and paternal protectiveness are viewed as intrinsic and developing critical factors that promote resiliency and motivate behavioral change. PMID- 17403494 TI - Research brief: sexual communication and knowledge among Mexican parents and their adolescent children. AB - This study describes the sexual knowledge and communication of Mexican parents and adolescents. Preintervention data were analyzed from 829 high school students (ages 14-17) and one of the parents of each. Differences were found between parents and adolescents in sexual knowledge (M = 16.16 vs. M = 14.92; t = 7.20, p < .001); specifically, parents had higher knowledge related to sexually transmitted diseases, HIV/AIDS, and condom use. Parents perceived more general communication (t [787] = 6.33, p < .001), and less discomfort talking about sex (t [785] = 4.69, p < .001) than adolescents. Parents with higher education levels scored higher in HIV knowledge and general communication. Fathers had higher total sexual knowledge, whereas mothers perceived higher sexual communication than fathers. There were no differences in knowledge and communication by parental socioeconomic level. Results suggest health care providers need to assist parents in developing specific knowledge and skills to support their adolescents' sexual decision-making. PMID- 17403495 TI - Culture-specific factors contributing to HIV risk among Jamaican adolescents. AB - The purpose of this article is to describe the results from an elicitation research study addressing the multisystem-level factors that contribute to HIV risk among Jamaican adolescents. Focus group and survey data were determined from parents, adolescents, and teachers in Kingston, Jamaica, from 2004 and 2005. Guided by an ecological extension of the Theory of Planned Behavior, focus groups and survey questionnaires identified cultural factors at the individual, family, and societal levels that significantly influence Jamaican adolescents' behavioral, normative, and control beliefs related to sexual behaviors that contribute to risk for HIV and other sexually transmitted infections. Although some factors were similar to those reported among adolescents living in the United States, others were culture-specific influences and beliefs that were unique to Jamaica. Results from the current study could contribute to the development of theory-based, culture-specific HIV risk-reduction interventions for use with Jamaican adolescents. PMID- 17403496 TI - Sexual stigma, sexual behaviors, and abstinence among Vietnamese adolescents: implications for risk and protective behaviors for HIV, sexually transmitted infections, and unwanted pregnancy. AB - As rates of HIV increase in Vietnam, there is a need for data on social relations and sexual risk and protective behaviors among Vietnamese adolescents in a context of rapid social and economic changes. The authors report results from qualitative interviews with 159 Vietnamese adolescents living in Hanoi, Nha Trang City, and Ninh Hoa District and a survey of 886 adolescents in these same three sites. In the qualitative interviews, youths report a strong adherence to ideals and values regarding abstinence outside of marriage. Youths reported low rates of engagement in vaginal, anal, and/or oral sex with a significant difference in reported behaviors between males (29/469, 6.2%) and females (7/416, 1.7%; p = .000). A total of 15 of 32 (46.9%) sexually active youths reported rarely or never using condoms. Females had significantly higher scores for perceived sexual stigma than males (t = -10.22 [95% confidence interval (CI); -3.72 to -2.52; p = .000) whereas males scored significantly lower than females on a scale of perceived self-efficacy for abstinence (t = 5.31 [95% CI; .27 to .59]; p = .000). The stigmatization of sexual relations outside of marriage particularly for young women reinforces abstinence; however, these same values decrease adolescents' ability to obtain accurate information about sexuality and HIV and sexually transmitted infections and engage in safer sex. PMID- 17403497 TI - Early adolescent peer leader development in HIV prevention using youth-adult partnership with schools approach. AB - This article describes the development and evaluation of an HIV prevention program developed for early adolescents through participatory action research. The HIV prevention program included a curriculum that was delivered by trained younger youth leaders through a youth and adult partnership with 10 schools in Chiang Mai Province, Thailand. The curriculum used participatory learning experiences, "edutainment" approaches, and skills-building strategies for enhancing youth leaders' capacities. Results of the evaluation showed that the senior-junior peer education program was effective in leadership role preparation, in improving youth leaders' ability to share sexual and reproductive health knowledge, and in promoting positive attitudes toward themselves. Success also rested on the fact that adults took a critical role in providing the opportunities, assistance, and guidance so that young people could develop their leadership capacity in an atmosphere of trust and respect. PMID- 17403498 TI - Research brief: community consultation to develop an acceptable and effective adolescent HIV prevention intervention. AB - HIV prevention for adolescents is urgently needed in Africa, but interventions have been slow to develop because of controversies about sex education. In this report the authors describe a four-step process used to develop a culturally and developmentally appropriate adolescent HIV prevention program for communities in rural Malawi. This is the final component of a 2-year ongoing adult HIV prevention program in Malawi. First the authors identified the risky behaviors of rural adolescents as well as cultural, developmental, and contextual factors. Next they consulted the community regarding how to use this information effectively and acceptably. Then an existing intervention was adapted based on this information. Finally, the authors piloted the intervention and made modifications based on lessons learned. This process provides a systematic way to consult with the community, thereby jointly enriching understanding, engaging the issues, and promoting support for an intervention program. PMID- 17403499 TI - Practice brief: adolescents and HIV clinical trials: ethics, culture, and context. AB - One quarter of HIV infections globally occur among young people 15 to 24 years of age, and more than half of all new infections are in people younger than 25 years. Clearly, there is a need to identify and implement effective HIV prevention strategies among at-risk teens. Some of the most effective options for slowing the epidemic are biomedical, and several promising methods are in development, including microbicides, vaccines, and preexposure prophylaxis (PREP, or the daily use of antiretrovirals to prevent the acquisition of HIV). There is widespread reluctance to enroll minors in such biomedical prevention trials because of concerns about vulnerability related to physical maturity, experiential maturity, and diminished autonomy as well as legal and social challenges that vary across and within nations. However, excluding minors from trials misses an important opportunity to evaluate the effectiveness, acceptability, and safety of innovative interventions under the best conditions for identifying and resolving potential problems. The challenges of including minors in HIV prevention trials are highlighted through the example of one rural South African community that has been particularly devastated by the HIV epidemic. PMID- 17403500 TI - Clinical decision support complements evidence-based decision making in dental practice. AB - Dental professionals as well as consumers of dental health care are driving the demand for access to reliable information so they can make more informed decisions. Clinical decision support (CDS) includes a variety of printed and electronic tools, systems, products, and services that make knowledge and information available to the user. CDS is the main way people will be able to access important facts, ideas, concepts, and the latest thinking about personal and population-based health subjects. CDS has its greatest potential at the point of care where it can facilitate good-quality evidence-based decision-making. PMID- 17403501 TI - Home tooth whitening systems improve tooth color over short term but lack evaluation of side effects. PMID- 17403502 TI - Ti-unite dental implant surface may be superior to machined surface in replacement of failed implants. PMID- 17403503 TI - Smokers are less likely to be aware of oral health effects of smoking than nonsmokers. PMID- 17403504 TI - Glass ionomer pit and fissure sealant provides caries protection on occlusal surfaces. PMID- 17403505 TI - Patients with acute painful TMD at high risk for developing a chronic condition report less pain, emotional distress, and health care use after a psychological intervention using cognitive-behavioral skills training and biofeedback. PMID- 17403506 TI - Rapid maxillary expansion results primarily in dental, not skeletal, changes. PMID- 17403507 TI - Synthetic platelet-derived growth factor (rhPDGF) may improve bone fill over beta tricalcium phosphate (beta-TCP) alone. PMID- 17403508 TI - A targeted program of providing 400-ppm F or 1450-ppm F toothpaste for low socioeconomic families was not associated with increased risk of esthetically objectionable fluorosis. PMID- 17403509 TI - Fluoride varnish application prevents caries in preschool children. PMID- 17403510 TI - Early mandibular first molar removal because of caries facilitates eruption of the ipsilateral third molar. PMID- 17403511 TI - Edentulous motherhood is associated with child caries. PMID- 17403512 TI - Contextual effect of socioeconomic status influences chronic periodontitis. PMID- 17403513 TI - Hospital emergency visits and admissions associated with dental problems increased by 121% between 1997 and 2001. PMID- 17403514 TI - Low socioeconomic position is associated with periodontitis in chilean adolescents. PMID- 17403515 TI - Quality of life of schoolchildren may be affected by poor alignment of their teeth. PMID- 17403516 TI - Hypodontia is associated with delayed dental development. PMID- 17403518 TI - Does magical thinking produce neutralising behaviour? An experimental investigation. AB - Magical thinking is of relevance to obsessive compulsive disorder (OCD), and has been most widely investigated in relation to the cognitive bias known as thought action fusion (TAF). This is seen as playing a role in the formation of fears about responsibility for harm. We suggest that magical thinking may also characterise some types of neutralising behaviour, which arise in response to those fears, and are a hallmark of the disorder. In an experimental study of 51 undergraduate students, we assessed whether the use of neutralising behaviours in response to an induction of fears of increasing likelihood for harm is related to a propensity for magical thinking. The 75.5% of participants demonstrated at least one form of neutralising behaviour in response to a TAF-induction task. Neutralising was associated with stronger and more persistent responses to the task, and with questionnaire measures of magical ideation. Those who neutralised did not report higher levels of OCD symptoms. It appears that neutralising is a common response in circumstances that provoke a sense of responsibility for harm. Its occurrence may be linked to magical thinking, however, the results from this experimental investigation suggested that this process may not be specific to OCD. PMID- 17403519 TI - Dihydrolipoic acid inhibits skin tumor promotion through anti-inflammation and anti-oxidation. AB - alpha-Lipoic acid (LA) has been intensely investigated as a therapeutic agent for several diseases, including hepatic disorder and diabetic polyneuropathy. However, the effects of LA or its reduced form, dihydrolipoic acid (DHLA), on cancer chemoprevention has never been reported. In the present study, we examined the effects of DHLA/LA on the production of nitric oxide (NO) by inducible NO synthase (iNOS) and the formation of prostaglandin E2 (PGE(2)) by cyclooxygenase 2 (COX-2), two important mediators associated with inflammation. DHLA/LA significantly inhibited lipopolysaccharide (LPS)-induced NO and PGE(2) formation in RAW 264.7 cells. Meanwhile, treatment with DHLA/LA suppressed the expression of iNOS protein but, unexpectedly, did not affect or increase the expression of COX-2 protein. The in vivo anti-inflammatory and antitumor-promoting activities were evaluated by a topical 12-O-tetradecanoylphorbol 13-acetate (TPA) application to mouse skin with measurement of edema formation, epidermal thickness and hydrogen peroxide production. DHLA significantly inhibited the priming and activation stages of skin inflammation induced by a double TPA application, by decreasing the inflammatory parameters. Furthermore, DHLA inhibited DMBA (0.3 micromol)/TPA (2.0 nmol)-induced skin tumor formation by reducing the tumor incidence and tumor multiplicity. When applied topically onto the shaven backs of mice prior to TPA, DHLA markedly inhibited the expression of iNOS protein. DHLA also strongly and directly inhibited COX-2 activity. These results suggest that DHLA can be a possible chemopreventive agent in inflammation associated tumorigenesis. PMID- 17403520 TI - Prevention of prostate cancer through custom tailoring of chemopreventive regimen. AB - One practical way to control cancer is through chemoprevention, which refers to the administration of synthetic or naturally occurring agents to block, reverse or delay the process of carcinogenesis. For a variety of reasons, the most important of which is human acceptance, for chemopreventive intervention naturally occurring diet-based agents are preferred over synthetic agents. For a long time, the prevailing mantra of cancer chemoprevention has been: "Find effective agents with acceptable or no toxicity and use them in preventing cancer in relatively healthy people or individuals at high risk for developing cancer". In pursuing this goal many naturally occurring phytochemicals capable of affording protection against carcinogenesis in preclinical settings in experimental animals have been described. However, clinical trials of single agents have yielded disappointing results. Since carcinogenesis is a multistage phenomenon in which many normal cellular pathways become aberrant, it is unlikely that one agent could prove effective in preventing cancer. This review underscores the need to build an armamentarium of naturally occurring chemopreventive substances that could prevent or slow down the development and progression of prostate cancer. Thus, the new effective approach for cancer prevention "building a customized mechanism-based chemoprevention cocktail of naturally occurring substances" is advocated. PMID- 17403521 TI - A diagnostic software tool for determination of complexity in respiratory pattern parameters. AB - The development and validation of a software that is user friendly and flexible in determining approximate entropy and reflecting complexity in respiratory pattern parameters are presented. The report includes the theory and computational methods for approximate entropy as well as the system description and software architecture. Results for a simulated periodic and regular respiratory pattern as well as for an irregular and complex breathing pattern obtained from a patient receiving mechanical ventilation in the intensive care unit are provided. By providing easy and rapid determination of the approximate entropy, the software enables health care professionals to understand how specific mechanical ventilation settings influence the respiratory pattern for patients receiving mechanical ventilation in the intensive care unit and ultimately identify the reversibility of respiratory diseases and weaning and liberation from mechanical ventilation. PMID- 17403522 TI - Image guidance in pelvic and acetabular surgery--expectations, success and limitations. AB - During the last decade navigation techniques in pelvic and acetabular surgery have been described. Nowadays, available techniques include CT-based navigation, 2D C-arm navigation and 3D C-arm navigation. The main indication is the navigated percutaneous SI screw fixation, but acetabular screw fixations are also reported. In this article, based upon a literature review and our own clinical experiences, the indications for and limitations of navigated techniques in pelvic and acetabular surgery are described. PMID- 17403523 TI - Optimization of in-situ monolithic synthesis for immunopreconcentration in capillary. AB - Poly(glycidyl methacrylate-co-ethylene dimethacrylate) monoliths have been synthetized in fused-silica capillary. The monomer mixture composition, initiation mode and porogen composition were optimized in order to provide a monolith with an homogeneous morphology and able to generate an electroosmotic flow via the incorporation of a small percentage of monomers possessing sulfonate group. Anti-ochratoxin A antibodies were immobilized through a single step on the epoxy groups leading to a miniaturized immunoextraction column. In order to evaluate the specificity of the analyte-antigen interaction on this immunosorbent, the retention of ochratoxin A was examined on this support but also on two complementary sorbents: one constituted by the non-bonded monolith and another one bonded with non-specific antibodies. Only the monolith bonded with anti-ochratoxin A antibodies lead to retention, showing the specificity of the interactions involved. This affinity phase based on a monolithic polymer support exhibits a high potential for specific preconcentration of small molecules. PMID- 17403524 TI - Factors affecting growth in the koinobiont endoparasitoid Venturia canescens in the flour moth Ephestia kuehniella. AB - With resistance of insect pests to synthetic pesticides on the increase, the role of parasitoid wasps as biological control agents is expanding in pest and resistance management strategies. One of the predictors of reproductive success of endoparasitoids is the relative size of the wasp at host emergence. While in idiobiont parasitoids, where the host stops feeding after parasitism, the wasp size is determined by the host size at the time of parasitism; the size of koinobiont wasps, where the host continues to feed after parasitism, is dependent on additional factors. Here we show that the host mass and temperature are important factors that determine survival and development of the koinobiont endoparasitoid Venturia canescens in late instar larvae of the flour moth Ephestia kuehniella. PMID- 17403525 TI - Assessment of oxidative DNA damage formation by organic complex mixtures from airborne particles PM(10). AB - The free radical generating activity of airborne particulate matter (PM(10)) has been proposed as a primary mechanism in biological activity of ambient air pollution. In an effort to determine the impact of the complex mixtures of extractable organic matter (EOM) from airborne particles on oxidative damage to DNA, the level of 8-oxo-2'-deoxyguanosine (8-oxodG), the most prevalent and stable oxidative lesion, was measured in the human metabolically competent cell line Hep G2. Cultured cells were exposed to equivalent EOM concentrations (5 150microg/ml) and oxidative DNA damage was analyzed using a modified single cell gel electrophoresis (SCGE), which involves the incubation of whole cell DNA with repair specific DNA endonuclease, which cleaves oxidized DNA at the sites of 8 oxodG. EOMs were extracted from PM(10) collected daily (24h intervals) in three European cities: Prague (Czech Republic, two monitoring sites, Libus and Smichov), Kosice (Slovak Republic) and Sofia (Bulgaria) during 3-month sampling periods in the winter and summer seasons. No substantial time- and dose-dependent increase of oxidative DNA lesions was detected in EOM-treated cells with the exception of the EOM collected at the monitoring site Kosice, summer sampling. In this case, 2h cell exposure to EOM resulted in a slight but significant increase of oxidative DNA damage at three from total of six concentrations. The mean 8 oxodG values at these concentrations ranged from 15.3 to 26.1 per 10(6) nucleotides with a value 3.5 per 10(6) nucleotides in untreated cells. B[a]P, the positive control, induced a variable but insignificant increase of oxidative DNA damage in Hep G2 cell (approximately 1.6-fold increase over control value). Based on these data we believe that EOM samples extracted from airborne particle PM(10) play probably only a marginal role in oxidative stress generation and oxidative lesion formation to DNA. However, adsorbed organic compounds can undergo various interactions (additive or synergistic) with other PM components or physical factors (UV-A radiation) and in this way they might enhance/multiply the adverse health effects of air pollution. PMID- 17403526 TI - Biomarkers of exposure to carcinogenic PAHs and their relationship with environmental factors. AB - The EXPAH project is a multicentre European study in which biomarkers of exposure, biomarkers of effect, genetic susceptibility and environmental factors were studied in populations exposed to differing levels of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs). We describe here the relationships between the levels of DNA adducts (as biomarkers of exposure), the exposure to air pollution and smoking status. Lymphocyte bulky DNA adducts were significantly correlated with exposure when subjects were classified either by job description or by personal monitor measurements, and both bulky and benzo(a)pyrene (B[a]P) DNA adducts were also correlated with smoking status. These associations varied across the countries studied (Czech Republic, Slovakia, Bulgaria). Results from a multivariate analysis show that factors mainly contributing to bulky and B[a]P DNA adducts are age, smoking habit, country of origin and environmental exposure to c-PAHs. The B[a]P DNA adducts were more strongly associated with smoking status than with the environmental exposure to c-PAHs. PMID- 17403527 TI - p53 codon 72 polymorphism, DNA damage and repair, and risk of non-melanoma skin cancer. AB - A very common polymorphism of p53, that of codon 72, codes either for a proline (P72) or an arginine (R72). The two alleles differ in their biological properties: P72 is a stronger inducer of p21, while R72 induces 5-10 times more apoptosis. It is not known, however, whether this polymorphism influences genome stability. The influence of p53 codon 72 polymorphism on cancer risk has been studied for different types of cancer with mixed and inconsistent results. With respect to sporadic non-melanoma skin cancer (NMSC), there are few studies, with small sample sizes, and none in a Latinoamerican population. These studies have found no association between p53 genotype at codon 72 and NMSC. We analyzed whether p53 codon 72 genotype influences genomic stability and the sensitivity of cells to UVB. We also carried out a case-control study of NMSC in a Mexican population which included 204 BCC cases, 42 SCC cases, and 238 controls. There was no association between p53 genotype and basal levels of DNA damage, oxidative DNA damage sensitivity, or DNA repair capacity. R72 dominantly increased the in vitro sensitivity of cells to UVB-induced apoptosis. There was no significant association either between p53 genotype and basal cell carcinoma (BCC), squamous cell carcinoma (SCC) or both combined. PMID- 17403528 TI - Effects of metabolic genotypes on intermediary biomarkers in subjects exposed to PAHS: results from the EXPAH study. AB - Data from the EXPAH project on PAH exposure and intermediary biomarkers were analyzed with respect to individual genotypes at seven metabolic gene loci. The GSTM1 null allele was associated with significantly higher levels of two biomarkers, malondialdehyde-2'-deoxyguanosine and benzo[a]pyrene DNA adducts in the total population from three Central and Eastern European countries. The CYP1B1 Leu/Val variant demonstrated effects on both markers of oxidative DNA damage in opposite directions, producing a higher level of M(1)dG with a trend from wild type (Leu/Leu) to heterozygotes to homozygous (Val/Val) variants, whereas the effects of these variants were reversed for 8-oxodG. Cluster Analysis was used to group composite genotypes in order to determine if combined genotypes of multiple loci could explain some of the variation seen with the biomarkers, expressed per unit of exposure, referred to as a sensitivity index. This analysis revealed two closely related genotypes each involving four of the loci (GSTM1*0/*0, CYP1A1*1*1, CYP1B1*1/*2, GSTP1*1/*1 and GSTT1*0/*0, CYP1A1*1*1, CYP1B1*1/*2, GSTP1*1/*1.) that conferred significant resistance to the DNA damaging effects of benzo[a]pyrene, measured as the level of a benzo[a]pyrene like adduct per unit of benzo[a]pyrene exposed. PMID- 17403529 TI - Neural representation of binding lexical signs and words in the episodic buffer of working memory. AB - The episodic buffer accommodates formation and maintenance of unitary multidimensional representations based on information in different codes from different sources. Formation, based on submorphemic units, engages posterior brain regions, while maintenance engages frontal regions. Using a hybrid fMRI design, that allows separate analysis of transient and sustained components, an n back task and an experimental group of 13 hearing native signers, with experience of Swedish Sign Language and Swedish since birth, we investigated binding of lexical signs and words in working memory. Results show that the transient component of these functions is supported by a buffer-specific network of posterior regions including the right middle temporal lobe, possibly relating to binding of phonological loop representations with semantic representations in long-term memory, as well as a loop-specific network, in line with predictions of a functional relationship between loop and buffer. The left hippocampus was engaged in transient and sustained components of buffer processing, possibly reflecting the meaningful nature of the stimuli. Only a minor role was found for executive functions in line with other recent work. A novel representation of the sustained component of working memory for audiovisual language in the right inferior temporal lobe may be related to perception of speech-related facial gestures. Previous findings of sign and speech loop representation in working memory were replicated and extended. Together, these findings support the notion of a module that mediates between codes and sources, such as the episodic buffer, and further our understanding of its nature. PMID- 17403530 TI - Auditory cortex and anterior cingulate cortex sources of the early evoked gamma band response: relationship to task difficulty and mental effort. AB - High frequency oscillations in the 40 Hz (gamma-band)-range are involved in the synchronization of brain regions, e.g., in cognitive functions. It has been suggested that the auditory evoked gamma-band response (GBR) is affected by attention and apart from auditory cortex activity a frontal or anterior cingulate cortex (ACC) generator could be involved. It was the aim of the present study to address three questions: (1) is there a neural generator of the early evoked GBR in the dorsal (d)ACC? (2) Are there different activation patterns in the dACC and the auditory cortex areas in response to task difficulty? (3) Is it possible to detect an influence of early ACC-gamma-band activity (GBR timeframe) to later auditory information processing (N1 timeframe)? In the present EEG/ERP-study we have investigated 30 healthy subjects using six auditory reaction tasks with increasing difficulty and mental effort demands. In the MANOVA analysis we found a significant main effect of task difficulty on both the GBR amplitude (F=7.75; p<0.001) and the auditory evoked N1 potential (F=7.00; p<0.001) with higher amplitudes in the more difficult tasks. In the LORETA region of interest (ROI) analysis, this effect was only due to increased dACC-activity during the GBR timeframe. For the ROI analysis during the N1 timeframe, in addition to a strong effect of task difficulty in the dACC a similar main effect was found in the auditory association area 22. These findings are in line with a top-down influence of dACC-activity to the auditory association area 22 during the early evoked GBR. PMID- 17403531 TI - Isolation and characterization of a novel P-II class snake venom metalloproteinase from Trimeresurus stejnegeri. AB - Stejnitin, a novel class P-II snake venom metalloproteinase (SVMP) with a molecular weight of about 35kDa, was purified from Trimeresurus stejnegeri venom. The cDNA of stejnitin encoded a polypeptide of 295 amino acid residues which comprises a signal peptide, proprotein, metalloproteinase domain, spacer and disintegrin domain. The protein sequence deduced from cDNA was confirmed by peptide mass fingerprinting analysis. It is highly homologous to the members of subclass P-IIa SVMPs which comprises metalloproteinase and disintegrin together. Results from DNA fragmentation and flow cytometry analysis also indicated that stejnitin is able to induce apoptosis of ECV304 cells (R=0.908, P=0.012). PMID- 17403532 TI - Pregnancy diagnosis in dairy cows by whey progesterone analysis: an ROC approach. AB - Concentration of progesterone in milk may be used to predict pregnancy status of dairy cattle by the 21st day after insemination. However, the accuracy of this method may be affected by fat-solubility of progesterone and sample storage conditions. After coagulation of a milk sample with rennet, an alternative method is to quantify progesterone concentration in whey with a novel, validated EIA. In this experiment, a receiver operating characteristic (ROC) analysis was performed to estimate the optimal discrimination point for whey progesterone concentration, using a sample of 991 Friesian cows evaluated between the 42nd and 44th day after insemination. Cows also were diagnosed for pregnancy by rectal palpation at this time. The overall conception rate at palpation was 57%. ROC analysis indicated that 259 pg/mL progesterone in whey was the most effective cutoff to discriminate correctly between pregnant and non-pregnant cows. Using this point for prediction, sensitivity was 98.2%, specificity was 70.9% and the area under ROC curve was 0.859, levels generally considered to denote moderate accuracy. The negative likelihood ratio at the cutoff of 259 pg/mL was 0.02, indicating satisfactory performance in detecting negative subjects, while the positive likelihood ratio (+LR=3.37) suggested average performance. In conclusion, EIA of progesterone concentration in whey is a viable method for predicting pregnancy status in cows. However, operators should take management objectives for the herd into account in determining the cutoff point and also considering important influencing variables such as conception rate in the herd. This method can provide diagnostic support for efforts to improve reproductive success, especially in low-fertility herds. PMID- 17403533 TI - Corpus luteum function and embryonic mortality in buffaloes treated with a GnRH agonist, hCG and progesterone. AB - The effect of treatment with a GnRH agonist, hCG or progesterone (P(4)) on corpus luteum function and embryonic mortality was investigated in buffaloes inseminated during mid-winter. Italian Mediterranean buffaloes (n=309) were synchronized using the Ovsynch with timed-AI program and mated by AI at 16 h (Day 0) and 40 h after the second injection of GnRH. On Day 5, buffaloes were randomly assigned to four groups: Control (no treatment, n=69), GnRH agonist (buserelin acetate, 12.6 microg, n=73), hCG (1500 IU, n=75) and P(4) (PRID without E(2) for 10 days, n=77). Progesterone (pg/ml) was determined in milk whey on Days 5, 10, 15 and 20 and pregnancy diagnosis was undertaken on Day 26 by ultrasound and Day 40 by rectal palpation. Treatment with buserelin and hCG increased (p<0.05) P(4) on Day 15 compared with controls (456+/-27, 451+/-24 and 346+/-28 pg/ml, respectively). Buffaloes treated with a PRID had intermediate P(4) concentrations (380+/-23 pg/ml). Embryonic mortality between Days 26 and 40 (22.9%) and pregnancies at Day 40 (48.9%) did not differ between treatments. A higher (p<0.01) P(4) concentration was found on Day 20 in pregnant animals compared with non-pregnant and embryonic mortality buffaloes, which did not differ. In summary, buserelin and hCG increased P(4) concentrations on Day 15 but this was not associated with a reduced incidence of embryonic mortality in buffaloes during mid-winter. PMID- 17403534 TI - Studies of the interaction between paraquat and bovine hemoglobin. AB - The interaction between paraquat (PQ) and bovine hemoglobin (BHb) was investigated using fluorescence and UV/vis absorption spectroscopy. The reactivity of the heme centers with superoxide anions formed by PQ was judged on the basis of the decrease of the Soret band. The experimental results showed that the fluorescence quenching of BHb by PQ was a result of the formation of PQ-BHb complex; static quenching was confirmed to result in the fluorescence quenching. The binding site number n, apparent binding constant K(A) and corresponding thermodynamic parameters were measured at different temperatures. The process of binding PQ molecule on BHb was a spontaneous molecular interaction procedure in which entropy increased and Gibbs free energy decreased. Hydrophobic and electrostatic interactions played a major role in stabilizing the complex. The effect of PQ on the conformation of BHb was analyzed using synchronous fluorescence spectroscopy. PMID- 17403535 TI - Comparative gene expression analysis of a chronic myelogenous leukemia cell line resistant to cyclophosphamide using oligonucleotide arrays and response to tyrosine kinase inhibitors. AB - Acquired imatinib resistance in chronic myelogenous leukemia (CML) can be the consequence of mutations in the kinase domain of BCR-ABL or increased protein levels. However, as in other malignancies, acquired resistance to cytostatic drugs is a common reason for treatment failure or disease progression. As a model for drug resistance, we developed a CML cell line resistant to cyclophosphamide (CP). Using oligonucleotide arrays, we examined changes in global gene expression. Selected genes were also examined by real-time PCR and flow cytometry. Neither the parent nor the resistant lines had mutations in their ATP binding domain. Filtering genes with a low-base line expression, a total of 239 genes showed significant changes (162 up- and 77 down-regulated) in the resistant clone. Most of the up-regulated genes were associated with metabolism, signal transduction, or encoded enzymes. The gene for aldehyde dehydrogenase 1 was over expressed more than 2000-fold in the resistant clone. BCR-ABL was expressed in both cell lines to a comparable extent. When exposed to the tyrosine kinase inhibitors imatinib and nilotinib, both lines were sensitive. In conclusion, we found multiple genetic changes in a CML cell line resistant to CP related to metabolism, signal transduction or apoptosis. Despite these changes, the resistant cells retained sensitivity to tyrosine kinase inhibitors. PMID- 17403536 TI - Effects of AMD3100 on transmigration and survival of acute myelogenous leukemia cells. AB - Acute myelogenous leukaemia (AML) blasts transmigrate in response to SDF-1alpha. AMD3100, a novel bicyclam molecule which inhibits stromal-derived factor (SDF) 1alpha/CXCR4 interactions, inhibited the transmigration of AML blasts and inhibited outgrowth of leukemia colony forming units. AMD3100 did not abrogate stroma-mediated protection from cytarabine-mediated apoptosis, except in the case of one promyelocytic leukemic sample tested, and it did not influence adhesion of blasts to endothelial monolayers. When AML blasts were pretreated with AMD3100, the positive effects of SDF-1alpha on NOD/SCID engraftment were diminished. This work confirms that AML is influenced by the SDF-1alpha/CXCR4 axis and demonstrates that disruption of this axis by the bicyclam AMD3100 can influence AML microenvironmental interactions. PMID- 17403537 TI - Bone marrow necrosis related to imatinib mesylate therapy for cml bilineal blast crisis. PMID- 17403538 TI - Various human epithelial cells express functional Toll-like receptors, NOD1 and NOD2 to produce anti-microbial peptides, but not proinflammatory cytokines. AB - Epithelial cells may form the first barrier of defense against bacteria in human tissues. We recently revealed that oral epithelial cells generated anti-bacterial factors, such as peptidoglycan recognition proteins (PGRPs) and beta-defensin 2, but not proinflammatory cytokines, such as interleukin-8 (IL-8), upon stimulation with bacterial cell-surface components. In this study, we found clear expressions of Toll-like receptor (TLR)2, TLR3, TLR4, TLR7, NOD1 and NOD2 in oral, tongue, salivary gland, pharyngeal, esophageal, intestinal, cervical, breast, lung, and kidney epithelial cells. However, tongue, salivary gland, pharyngeal, esophageal, intestinal, cervical, breast, lung, and kidney epithelial cells, as well as oral epithelial cells, did not secrete IL-6, IL-8 or monocyte chemoattractant protein 1 in response to chemically synthesized TLR and NOD agonists mimicking microbial components: TLR2 agonistic lipopeptide (Pam3CSSNA), TLR3 agonistic Poly I:C, TLR4 agonistic lipid A (LA-15-PP), TLR7 agonistic single stranded RNA (ssPoly U), NOD1 agonistic iE-DAP (gamma-D-glumtamyl-meso-diaminopimelic acid), and NOD2 agonistic muramyldipeptide (MDP). Although PGRPs on oral epithelial cells were significantly up-regulated upon stimulation with these synthetic components, PGRPs on pharyngeal epithelial cells were only slightly up-regulated, and PGRPs on esophageal, intestinal and cervical epithelial cells were not up-regulated upon stimulation with the components. In contrast, stimulation with synthetic TLRs and NODs ligands induced beta-defensin 2 generation in all epithelial cells examined. These findings indicate that TLR and NOD in various epithelial cells are functional receptors that induce anti-bacterial responses in general without being accompanied by inflammatory responses. PMID- 17403539 TI - 17 Beta-estradiol normalizes Toll receptor 4, mitogen activated protein kinases and inflammatory response in epidermal keratinocytes following trauma-hemorrhage. AB - Trauma-hemorrhage produces immunodepression in males but not in proestrus females and this difference is due to the presence of high estrogen in proestrus females. Although skin is the largest immunological organ of the body and is considered the first line of defense, no study to-date has examined whether trauma hemorrhage has any effects on keratinocytes which are the major epidermal cell type (>90%) of skin. We therefore examined whether epidermal keratinocytes inflammatory response and the signal transduction pathways involved in the inflammatory response are altered following trauma-hemorrhage. C3H/HeN mice were subjected to trauma-hemorrhage and 2h thereafter; keratinocytes were harvested and stimulated with LPS for 24h (5 microg/ml). Inflammatory mediators, Toll-like receptor (TLR) and myeloid differentiation adaptor protein (MyD88) expression, and the activation of mitogen-activated protein kinase (MAPK) were determined. Trauma-hemorrhage increased the production of IL-6, IL-10, IL-12 and TNF-alpha enhanced the expression of TLR4, MyD88 as well as the activation of MAPK proteins (p38, ERK and JNK) in epidermal keratinocytes. However, administration of a single dose of 17beta-estradiol following trauma-hemorrhage prevented the increase in these inflammatory parameters under those conditions. These findings suggest that 17beta-estradiol normalizes epidermal keratinocytes inflammatory responses following trauma-hemorrhage by preventing the upregulation of TLR4 mediated MAPK activation. PMID- 17403540 TI - TGF-beta 1 inhibition of IFN-gamma-induced signaling and Th1 gene expression in CD4+ T cells is Smad3 independent but MAP kinase dependent. AB - In addition to classic Smad signaling pathways, the pleiotropic immunoregulatory cytokine TGF-beta1 can activate MAP kinases, but a role for TGF-beta1-MAP kinase pathways in T cells has not been defined heretofore. We have shown previously that TGF-beta1 inhibits Th1 development by inhibiting IFN-gamma's induction of T bet and other Th1 differentiation genes, and that TGF-beta1 inhibits receptor proximal IFN-gamma-Jak-Stat signaling responses. We now show that these effects of TGF-beta1 are independent of the canonical TGF-beta1 signaling module Smad3, but involve a specific MAP kinase pathway. In primary T cells, TGF-beta1 activated the MEK/ERK and p38 MAP kinase pathways, but not the JNK pathway. Inhibition of the MEK/ERK pathway completely eliminated the inhibitory effects of TGF-beta1 on IFN-gamma responses in T cells, whereas inhibition of the p38 pathway had no effect. Thus, TGF-beta1's inhibition of IFN-gamma signaling in T cells is mediated through a highly specific Smad3 independent, MEK/ERK-dependent signaling pathway. PMID- 17403541 TI - Pepsin pretreatment allows collagen IV immunostaining of blood vessels in adult mouse brain. AB - While the brain vasculature can be imaged with many methods, immunohistochemistry has distinct advantages due to its simplicity and applicability to archival tissue. However, immunohistochemical staining of the murine brain vasculature in aldehyde fixed tissue has proven elusive and inconsistent using current protocols. Here we investigated whether antigen retrieval methods could improve vascular staining in the adult mouse brain. We found that pepsin digestion prior to immunostaining unmasked widespread collagen IV staining of the cerebrovasculature in the adult mouse brain. Pepsin treatment also unmasked widespread vascular staining with laminin, but only marginally improved isolectin B4 staining and did not enhance vascular staining with fibronectin, perlecan or CD146. Collagen IV immunoperoxidase staining was easily combined with cresyl violet counterstaining making it suitable for stereological analyses of both vascular and neuronal parameters in the same tissue section. This method should be widely applicable for labeling the brain vasculature of the mouse in aldehyde fixed tissue from both normal and pathological states. PMID- 17403542 TI - Transfer of maternal cytokines to suckling piglets: in vivo and in vitro models with implications for immunomodulation of neonatal immunity. AB - Maternal cytokines may play instructive roles in development of the neonatal immune system. However, cytokines in colostrum and milk and their transfer from mothers to neonates have not been well documented, except for TGF-beta. Swine provide a unique model to study lactogenic cytokines because the sow's impermeable placenta prohibits transplacental passage. We investigated IL-6 and TNF-alpha (pro-inflammatory), IFN-gamma and IL-12 (Th1), IL-10 and IL-4 (Th2) and TGF-beta1 (Th3) concentrations in sow serum and colostrum/milk and serum of their suckling and weaned piglets and in age-matched colostrum-deprived gnotobiotic piglets. All cytokines were detected in colostrum/milk and correlated with concentrations in sow serum except for mammary-derived TNF-alpha and TGF-beta1. Detection of IL-12 and TGF-beta1 in pre-suckling and colostrum-deprived gnotobiotic piglet serum suggests constitutive production: other cytokines were undetectable confirming absence of transplacental transfer. Peak median cytokine concentrations in suckling piglet serum occurred at post-partum days 1-2 (IL-4>IL 6>IFN-gamma>IL-10). The effects in vitro of physiologically relevant concentrations of the two predominant lactogenic cytokines (TGF-beta1 and IL-4) on porcine naive B cell responses to lipopolysaccharide (LPS) and rotavirus (RV) were investigated. High (10 ng/ml) TGF-beta1 suppressed immunoglobulin secreting cell responses to LPS and rotavirus; low concentrations (0.1 ng/ml) promoted isotype switching to IgA antibody. Interleukin-4 induced inverse dose-dependent (0.1 ng>10 ng/ml) isotype switching to IgA and enhanced IgM secreting cell responses to LPS and rotavirus. In summary, we documented the transfer and persistence of maternal cytokines from colostrum/milk to neonates and their potential role in Th-2 biased IgA responses and reduced immunologic responsiveness of neonates. PMID- 17403543 TI - Functional biology of the alpha(2)delta subunits of voltage-gated calcium channels. AB - In this review, we examine what is known about the mechanism of action of the auxiliary alpha2delta subunits of voltage-gated Ca(2+) (Ca(v)) channels. First, to provide some background on the alpha2delta proteins, we discuss the genes encoding these channels, in addition to the topology and predicted structure of the alpha2delta subunits. We then describe the effects of alpha2delta subunits on the biophysical properties of Ca(v) channels and their physiological function. All alpha2delta subunits increase the density at the plasma membrane of Ca(2+) channels activated by high voltage, and we discuss what is known about the mechanism underlying this trafficking. Finally, we consider the link between alpha2delta subunits and disease, both in terms of spontaneous and engineered mouse mutants that show cerebellar ataxia and spike-wave epilepsy, and in terms of neuropathic pain and the mechanism of action of the gabapentinoid drugs - small-molecule ligands of the alpha2delta-1 and alpha2delta-2 subunits. PMID- 17403544 TI - Development and evaluation of SYBR Green I-based one-step real-time RT-PCR assay for detection and quantitation of Japanese encephalitis virus. AB - One-step SYBR Green I-based real-time RT-PCR assay for rapid detection as well as quantitation of Japanese encephalitis virus (JEV) in acute-phase patient CSF samples by targeting the NS3 gene was developed. The assay developed in this study was found to be more sensitive as compared to conventional RT-PCR. The specificity of the reported assay system was established through melting curve analysis as well as by cross-reactivity studies with related members of Flavivirus. The applicability of Real-time PCR assay for clinical diagnosis was validated with 32 suspected acute-phase CSF samples of Gorakhpur epidemic, India, 2005. The improved sensitivity of real-time RT-PCR was reflected by picking up 10 additional samples with low copy number of template in comparison to conventional RT-PCR. The quantitation of the viral load in acute-phase CSF samples was done using a standard curve obtained by plotting cycle threshold (C(t)) values versus copy numbers of the RNA template. This is the first report on the application of real-time RT-PCR for detection as well as quantitation of JEV from patient CSF samples. These findings demonstrate the potential clinical application of the reported assay as a sensitive diagnostic test for rapid and real-time detection and quantitation of JEV in acute-phase clinical samples. PMID- 17403545 TI - Temporary inactivation of the dorsal hippocampus induces a transient impairment in retrieval of aversive memory. AB - The reconsolidation hypothesis, which predicts that consolidated memories become labile and sensitive to amnestic agents upon reactivation, has been one of the most debated topics in memory research over recent years. One of the main criticisms to this hypothesis is the fact that some studies have shown the effects of "reconsolidation blockade" to be transient. Here we show that muscimol inactivation of the dorsal hippocampus following memory reactivation produces a reversible impairment of step-down inhibitory avoidance memory in rats. Moreover, we show that the reversal of this effect is dependent on the passage of time, and not on repeated testing. The implications of the findings to the interpretation of the phenomenon of transient retrieval impairment induced by post-reactivation pharmacological interventions on memory systems are discussed. PMID- 17403546 TI - Identification of gene expression indicators for thyroid axis disruption in a Xenopus laevis metamorphosis screening assay. Part 1. Effects on the brain. AB - Thyroid hormones (TH), thyroxine (T(4)) and 3,5,3'-triiodothyronine (T(3)), play crucial roles in regulation of growth, development and metabolism in vertebrates and their actions are targets for endocrine disruptive agents. Perturbations in TH action can contribute to the development of disease states and the US Environmental Protection Agency is developing a high throughput screen using TH dependent amphibian metamorphosis as an assay platform. Currently this methodology relies on external morphological endpoints and changes in central thyroid axis parameters. However, exposure-related changes in gene expression in TH-sensitive tissue types that occur over shorter time frames have the potential to augment this screen. This study aims to characterize and identify molecular markers in the tadpole brain. Using a combination of cDNA array analysis and real time quantitative polymerase chain reaction (QPCR), we examine the brain of tadpoles following 96 h of continuous exposure to T(3), T(4), methimazole, propylthiouracil, or perchlorate. This tissue was more sensitive to T(4) rather than T(3), even when differences in biological activity were taken into account. This implies that a simple conversion of T(4) to T(3) cannot fully account for T(4) effects on the brain and suggests distinctive mechanisms of action for the two THs. While the brain shows gene expression alterations for methimazole and propylthiouracil, the environmental contaminant, perchlorate, had the greatest effect on the levels of mRNAs encoding proteins important in neural development and function. Our data identify gene expression profiles that can serve as exposure indicators of these chemicals. PMID- 17403547 TI - An Entamoeba sp. strain isolated from rhesus monkey is virulent but genetically different from Entamoeba histolytica. AB - An Entamoeba sp. strain, P19-061405, was isolated from a rhesus monkey in Nepal and characterized genetically. The strain was initially identified as Entamoeba histolytica using PCR amplification of peroxiredoxin genes. However, sequence analysis of the 18S rRNA gene showed a 0.8% difference when compared to the reference E. histolytica HM-1:IMSS human strain. Differences were also observed in the 5.8S rRNA gene and the internal transcribed spacer (ITS) regions 1 and 2, and analysis of the serine-rich protein gene from the monkey strain showed unique codon usages compared to E. histolytica isolated from humans. The amino acid sequences of two hexokinases and two glucose phosphate isomerases also differed from those of E. histolytica. Isoenzyme analyses of these enzymes in the monkey strain showed different electrophoretic mobility patterns compared with E. histolytica isolates. Analysis of peroxiredoxin genes indicated the presence of at least seven different types of protein, none of which were identical to proteins in E. histolytica. When the trophozoites from the monkey strain were inoculated into the livers of hamsters, formation of amebic abscesses was observed 7 days after the injection. These results demonstrate that the strain is genetically different from E. histolytica and is virulent. Revival of the name Entamoeba nuttalli is proposed for the organism. PMID- 17403548 TI - Hospital admissions among people 65+ related to multimorbidity, municipal and outpatient care. AB - This study aimed at examine the number of planned and acute hospital admissions during 1 year among people 65+ and its relation to municipal care, outpatient care, multimorbidity, age and sex. Four thousand nine hundred and seven individuals having one or more admissions during 2001 were studied. Data were collected from two registers and comparisons were made between those having one, two and three or more hospital stays and between those with and without municipal care and services. Linear regression was used to examine factors predicting number of acute and planned admissions. Fifteen percent of the sample had three or more hospital stays (range 3-15) accounting for 35% of all admissions. This group had significantly more contacts in outpatient care with physician (median number of contacts (md)=15), compared to those with one (md: 8), or two admissions (md: 11). Main predictors for number of admissions were number of diagnosis groups and number of contacts with physician in outpatient care. Those who are frequently admitted to hospital constitute a small group that consume a great deal of inpatient care and also tend to have frequent contacts in outpatient care. Thus interventions focusing on frequent admissions are needed, and this requires collaboration between outpatient and hospital care. PMID- 17403549 TI - Bioenergetic strategy of microalgae for the biodegradation of phenolic compounds: exogenously supplied energy and carbon sources adjust the level of biodegradation. AB - The biodegradation of phenolic compounds by microalgae seems to be not a simple feature of a particular organism, but mostly a bioenergetic process depending on the growth conditions, especially on the exogenously supplied energy (carbon and light) sources. By using chlorophyll fluorescence induction measurements to estimate the molecular structure and function of the photosynthetic apparatus and therefore the tolerance/sensitivity of microalgae incubated with phenols, it can be assumed that, at least in low concentrations, phenol have no toxic effects on the cultures and can be used as alternative carbon source in them. Halophenols (chlorophenols, bromophenols and iodophenols) are quite toxic for the microalgal cultures. In halophenols the first step of the biodegradation is the split of the halogen substituent (dehalogenation). This is strongly determined by the bond dissociation energy of the corresponding substituent and therefore the energetic requirement for the biodegradation of halophenols increases following the sequence: iodophenolor=40. Titres to the HPAI H5N1 virus followed a similar trend, but were lower (GMT: 61 (95% CI: 49-76); 61%>or=40). The breadth of the immune response was further demonstrated by measuring antibody titres against prototype strains of four antigenic clades of currently circulating H5N1 viruses. These data indicate that vaccination should be regarded as a beneficial component of the preventive measures (including increased bio-security and monitoring) that can be undertaken in zoos to prevent an outbreak of and decrease environmental contamination by HPAI H5N1 virus, while alleviating confinement measures. PMID- 17403560 TI - Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross reactivity. AB - Many influenza vaccines targeted to hemagglutinin (HA) show efficient immunogenicity for protecting subjects against influenza virus infection. Major antigenic changes to HA molecules can help influenza virus to develop resistance against HA-targeted vaccines. DNA vaccines encoding conserved antigens protect animals against diverse subtypes, but their potency requires further improvement. We generated a DNA-based nucleoprotein (NP)-targeted vaccine using an N-terminal mutant of NP (NPm) that efficiently localized in the cytoplasm, and examined the immune responses in mice immunized with NPm or wild-type (WT) NP DNA vaccine. Importantly, the NPm vaccine showed 1.5-2-fold higher immunogenicity than the WT NP vaccine in mice. Furthermore, NPm vaccination efficiently protected the mice against lethal challenge with influenza viruses and showed cross-reactivity toward heterologous viruses. Therefore, DNA-based vaccination with NPm may contribute to the development of protective immunity against diverse influenza virus through its ability to stimulate cellular immunity. PMID- 17403561 TI - Rash including mucosal involvement: case definition and guidelines for collection, analysis, and presentation of immunization safety data. PMID- 17403562 TI - Evaluation of influenza virus-like particles and Novasome adjuvant as candidate vaccine for avian influenza. AB - The development of safe and effective vaccines for avian influenza viruses is a priority for pandemic preparedness. Adjuvants improve the efficacy of vaccines and may allow antigen sparing during a pandemic. We have previously shown that influenza virus-like particles (VLPs) comprised of HA, NA, and M1 proteins represent a candidate vaccine for avian influenza H9N2 virus [Pushko P, Tumpey TM, Fang Bu, Knell J, Robinson R, Smith G. Influenza virus-like particles comprised of the HA, NA, and M1 proteins of H9N2 influenza virus induce protective immune responses in BALB/c mice. Vaccine 2005;23(50):5751-9]. In this study, an H9N2 VLP vaccine and recombinant HA (rH9) vaccine were evaluated in three animal models. The H9N2 VLP vaccine protected mice and ferrets from challenge with A/Hong Kong/1073/99 (H9N2) virus. Novasome adjuvant improved immunogenicity and protection. Positive effect of the adjuvant was also detected using the rH9 vaccine. The results have implications for the development of safe and effective vaccines for avian influenza viruses with pandemic potential. PMID- 17403563 TI - Fetal conditions and fatal decisions: ethical dilemmas in ultrasound screening in Vietnam. AB - In the context of globalization, new technologies of pregnancy are spreading rapidly from affluent to low-income countries. Yet, to date, there is very little research on the application of prenatal diagnostic technology in developing country settings or the dilemmas that prenatal screening may give rise to in situations where health-care resources are scarce. In this article, we describe how obstetrical ultrasound scanning is used as the most important technology for prenatal diagnosis in Vietnam. We explore the social context that shapes moral sentiments and ethical deliberations within everyday medical interactions, drawing attention to the complex social processes through which ethical dilemmas are configured. The data include observations in the scanning room, and semi structured interviews with patients and staff at a major maternity hospital in the country's capital Hanoi. We found that pregnant women received very limited information and support when a fetal anomaly was detected by ultrasound. Most women left the hospital feeling uncertain about what was wrong with the fetus and decision-making about abortion centred around the difficulties of parenting a disabled child in Vietnam. We conclude that the ethical problems surrounding prenatal screening are intensified in low-income settings such as Vietnam and point to the need for research that takes into account the wider social context that structures ethical dilemmas. PMID- 17403564 TI - Rhythm perception: Speeding up or slowing down affects different subcomponents of the ERP P3 complex. AB - The aim of this study was to investigate, by measuring the event related potential (ERP) P3 complex, whether the perception of small accelerations differs from that of small decelerations. Participants had to decide whether the last beat of a short sequence was presented 'too early' or 'too late'. Target beats were accelerated or decelerated with 0%, 2%, 5%, or 10%. Individuals differed in their capability to detect small tempo changes. We found that good responders were able to identify all tempo changes whereas poor responders were only able to identify large (10%) tempo changes. In addition, we found that tempo changes affected two subcomponents of the ERP P3 in good performers. Accelerations increased a late-P3 amplitude whereas decelerations increased an early-P3 amplitude. These results imply the principle possibility to measure differential P3 effects within one task. This is important for acquiring more refined knowledge concerning different subcomponents of the ERP P3 complex and the cognitive processes by which they are elicited. PMID- 17403565 TI - Voiding dysfunction after radical retropubic prostatectomy: more than external urethral sphincter deficiency. AB - OBJECTIVES: To analyse the relationship between RRP and urodynamic bladder dysfunction, and compare preoperative and postoperative functional status over long-term follow-up. Hypothesis on the pathophysiologic mechanism underlying urodynamic dysfunction has been reported. METHODS: PubMed databank search for original articles followed by review of urodynamic parameters: bladder filling sensation, detrusor overactivity, bladder compliance, cystometric bladder capacity, impaired detrusor contractility, bladder outlet obstruction, urinary incontinence. RESULTS: Detrusor dysfunction was rarely present as the sole diagnosis and was usually coupled with intrinsic sphincter deficiency. Data on bladder filling sensation, cystometric capacity, detrusor overactivity, impaired detrusor contractility, and bladder outlet obstruction were limited and contradictory. Detrusor overactivity was a de novo dysfunction in 2%-77% of patients. Impaired bladder compliance was present in 8%-39% of patients and was de novo in about 50%. Impaired detrusor contractility was found in 29%-61% of patients, was de novo in 47%, and recovered in about 50% of patients. The role of these dysfunctions as etiologic agents of urinary incontinence or voiding symptoms was unevenly assessed. CONCLUSIONS: Postoperative decentralization of the bladder, inflammation and/or infection, and geometric bladder wall alteration associated with preexisting hypoxemia with/without neuroplasticity have been posited as causes of detrusor dysfunction. Nevertheless, the lack of consistent preoperative urodynamic investigation makes it difficult to assess the operation's exact role in causing these dysfunctions. Thus, urodynamics performed, at least in selected cases, preoperatively and during follow-up could help arrive at a precise diagnosis of the underlying dysfunction, indicate the appropriate treatment, and prevent the incidence and onset of postoperative urinary incontinence. PMID- 17403566 TI - Electrochemical studies and square wave voltammetry of paraquat at natural phosphate modified carbon paste electrode. AB - A square wave voltammetry (SWV) method for the determination of trace amounts of paraquat at carbon paste electrode modified with natural phosphate (NP-CPE) is proposed. Paraquat was accumulated onto natural phosphate at open circuit potential from aqueous solution. The reduction peaks of paraquat were observed around -0.7V and -1.0V (versus SCE). The response of paraquat at NP-CPE related to: the concentration of this herbicide, preconcentration time, natural phosphate loading and measuring solution pH, was investigated. It was observed that the increase of the cathodic peak currents using SWV, under the optimized condition, is linear with the increase of paraquat concentration in the range from 2.3x10( 8)molL(-1) to 300x10(-8)molL(-1). The detection limit (DL, 3S.D.) and quantification limit (QL, 10S.D.) for peak 1 were about 7.8x10(-10)molL(-1) and 2.59x10(-9)molL(-1), respectively, in pure electrolyte water with the relative standard deviation of 1.8% (n=7). The proposed method was successfully applied to the determination of paraquat in real water samples with satisfactory results. PMID- 17403567 TI - Optimization of Fenton-biological treatment scheme for the treatment of aqueous dye solutions. AB - Degradation of dyes especially, azo dyes are difficult due to their complex structure and synthetic nature. The main objective of this study was to evaluate the Fenton-biological (aerobic) treatment train for decolorization and mineralization of azo dyes viz. Reactive Black 5 (RB5), Reactive Blue 13 (RB13) and Acid Orange 7 (AO7). The objective of Fenton treatment was only to decolorize the dyes (breakage of -NN-), as it was considered that after breakage of -NN-, the dyes will become amenable to biodegradation and can be further treated in aerobic biological system. Hence studies were carried out to optimize the lower Fenton's doses for decolorization of dyes. The optimum doses for decolorization (>95%) of all the three dyes were found out to be 15 mgL(-1) of Fe(2+) (0.27 mM) and 50 mgL(-1) (1.47 mM) of H(2)O(2) dose at optimum pH 3. Further it was also investigated that at lower doses, the main problem of Fenton process (sludge generation) can also be minimized. Later the mineralization of the dye (removal of aromatic amines) was achieved in the aerobic biological treatment system. Overall reduction of 64, 89 and 75% in the aromatic amines (at 254 nm), 88, 95 and 78% in naphthalene ring associated compounds (near 310 nm) and 49, 89 and 91% reduction in benzene ring associated compounds (near 226 nm) were observed for RB5, RB13 and AO7, respectively. Thus this treatment system seems to be quite effective and economical option for the treatment of recalcitrant compounds like dyes, as the cost in the chemical treatment is considered mainly due to chemicals thus at lower doses the operational cost is saved. Further, as the sludge generation was almost negligible at lower doses, thus the savings in cost of handling and disposal of hazardous sludge also adds to economy of treatment. PMID- 17403568 TI - Biosorption of Cr(VI) by immobilized biomass of two indigenous strains of cyanobacteria isolated from metal contaminated soil. AB - Biosorption of Cr(VI) using native strains of cyanobacteria from metal contaminated soil in the premises of textile mill has been reported in this paper. Biosorption was studied as a function of pH (1-5), contact time (5-180 min) and initial chromium ion concentration (5-20mg/l) to find out the maximum biosorption capacity of alginate immobilized Nostoc calcicola HH-12 and Chroococcus sp. HH-11. The optimum conditions for Cr(VI) biosorption are almost same for the two strains (pH 3-4, contact time 30 min and initial chromium concentration of 20mg/l) however, the biomass of Chroococcus sp. HH-11 was found to be more suitable for the development of an efficient biosorbent for the removal of Cr(VI) from wastewater, as it showed higher values of q(m) and K(f), the Langmuir and Freundlich isotherm parameters. Both the isotherm models were suitable for describing the biosorption of Cr(VI) by the cyanobacterial biosorbents. PMID- 17403569 TI - Expression of CD200 by macrophage-like cells in ischemic core of rat brain after transient middle cerebral artery occlusion. AB - Brain ischemia causes the death of neurons and glial cells. Such brain cells are believed to inevitably undergo degeneration in the core of ischemic lesions, whereas neurons and glial cells may survive in the region surrounding the core that is often referred to as the ischemic penumbra. However, many cells, particularly immune cells infiltrate and survive in the core. In this study, we characterized macrophage-like cells that accumulated in the ischemic core of a rat brain whose right middle cerebral artery was transiently occluded for 90 min. At 7 days post-reperfusion, we observed macrophage-like cells expressing CD200, a cell surface glycoprotein belonging to an immunoglobulin superfamily and that elicits suppressive effects on myeloid cells including microglia by interacting with the CD200 receptor (CD200R). RT-PCR and immunoblot analyses revealed the presence of CD200-mRNA and protein in the ischemic core as well as in the contralateral region. As revealed by immunohistochemistry, CD200 is located on the cell membrane of spherical Iba1(+) cells with many cytoplasmic granules. CD200(-)/Iba1(+) macrophage-like cells were also present, which have a more irregular shape than CD200(+)/Iba1(+) cells. CD200 was detected in isolated spherical Iba1(+) macrophage-like cells. Thus, CD200 is expressed in some populations of macrophage-like cells that may be responsible for the suppression of CD200R(+) myeloid cell functions in the ischemic core. PMID- 17403570 TI - Parkinsonian deficits in context-dependent regulation of standing postural control. AB - This study explored whether patients with Parkinson's disease alter the regulation of upright standing according to constraints imposed by the environmental context. The provision of context-dependent adaptations was inferred from the presence of adjustments to standing postural control that would serve to reduce fall risk when balance was challenged by a threatening environmental context. Participants were asked to stand as still as possible in two environmental context conditions that differed in the level of imposed postural threat: LOW threat and HIGH threat. Eight levodopa dependent patients with Parkinson's disease (PD) and eight age-matched control subjects (CTRL) provided the subject sample. PD patients were tested following a 12-h withdrawal of anti-Parkinsonian medications and approximately 1h post-medication. The CTRL group showed altered postural control in the HIGH threat condition, in a manner that was indicative of appropriate context-dependent regulation of standing. PD patients, in the non-medicated or medicated states, did not modify stance regulation when the environmental context heightened postural threat. Our results extend the current understanding of Parkinsonian deficits in the context dependent regulation of postural control to include upright standing. PMID- 17403571 TI - Effect of NMDA receptor antagonist on proliferation of neurospheres from embryonic brain. AB - The N-methyl-D-aspartate (NMDA) receptor, a subtype of ionotropic glutamate receptors, plays an important role in the regulation of neuronal development, learning and memory, and neurodegenerative diseases. NMDA receptor blockade enhances neurogenesis in the hippocampal dentate gyrus in vivo. The effect of NMDA receptor antagonist on proliferation of neural progenitor cells, however, remains to be determined. We investigated changes in the diameter and number of neurospheres derived from the embryonic rat brain after NMDA receptor blockade. Cortical progenitor cells were isolated from gestational day 18 fetal rats according to the Percoll density gradient method. Cultured spheres expressed neural progenitor markers, musashi-1 and nestin. Immunohistochemical analysis demonstrated that cells in Dulbecco's modified Eagle medium/F12 containing 1% fetal bovine serum on day 8 differentiated to MAP-2-positive neurons and GFAP positive astrocytes. The expression of NR1 and NR2B subunits of the NMDA receptor in neurospheres was detected. Neither brief nor sustained exposure to NMDA altered the diameter and number of neurospheres. Brief exposure to 30 microM MK 801, an NMDA receptor antagonist, decreased the diameter of neurospheres. Sustained exposure to 30 microM MK-801 decreased the diameter and number of neurospheres. Our results provide evidence that MK-801 directly decreased proliferation of neural progenitor cells. PMID- 17403572 TI - Acidic oligosaccharide sugar chain, a marine-derived oligosaccharide, activates human glial cell line-derived neurotrophic factor signaling. AB - Gial derived neurotrophic factor (GDNF) modulates neuronal cell differentiation during development and protects against neurodegeneration by preventing apoptosis at maturity. GDNF's role in tissue maintenance has generated interest in the therapeutic potential of GDNF in treating neurological disorders such as Parkinson's disease. Heparan sulfate has been shown to be essential for GDNF signaling and altering the levels of heparan sulfate promotes or inhibits GDNF functional activity. To search for other oligosaccharides capable of modulating GDNF activity as potential therapeutic molecules, we investigated the effect of acidic oligosaccharide sugar chain (AOSC) and its sulfated derivative on GDNF induced neurotrophic events by using Western-blotting, immunofluorescence cell staining, and immunoprecipitation techniques in PC12 cells expressing the GDNF receptors GFR alpha 1-Ret. AOSC significantly improved the neurite outgrowth and activated c-Ret phosphorylation in PC12-GFR alpha 1-Ret cells, but its sulfated derivative inhibited GDNF activity. Studies to understand the opposing biological effects of AOSC and its sulfated derivative on GDNF activity demonstrated that reduced GDNF binding to PC12-GFR alpha 1-Ret cell surface in the presence of the sulfated derivative likely suppressed GDNF activity as both AOSC and its sulfated derivatives had similar binding affinities to GDNF. This study illustrates the importance of oligosaccharide structure and charge on influencing GDNF activity and the potential use of oligosaccharides in modulating GDNF activity for therapeutic purposes. PMID- 17403573 TI - Differences of cortical activation pattern between cortical and corona radiata infarct. AB - The aim of this study was to elucidate the differences of cortical activation pattern between patients with cortex (CO) infarct and those with corona radiata (CR) infarct. Twenty chronic patients with cerebral infarct and 11 normal subjects were recruited. The patients were classified into two groups according to infarct location: CO and CR group. After functional MRI was performed during the motor task of hand grasp-release movements, the hand motor function was assessed. The CO group tended to be activated on the contralateral primary sensori-motor cortex, whereas the CR group largely showed activation of the bilateral primary sensori-motor cortex. Namely, there was significantly more lateralized activation pattern of primary sensori-motor cortex in the CO group than in the CR group. Additionally, the CO group had better hand motor function than the CR group. Our results suggest that the motor recovery mechanism vary according to the lesion location between CO and CR infarct, indicating that the adaptive cortical reorganization proceeds under different patterns according to the lesion location. PMID- 17403574 TI - Capillary beds are decreased in Alzheimer's disease, but not in Binswanger's disease. AB - Morphological abnormalities of the cortical microvessels have been reported in Alzheimer's disease (AD), but not in Binswanger's disease (BD), a form of vascular dementia. Therefore, we compared the capillary beds in AD and BD brains, using a modified Gallyas silver impregnation method and immunohistochemistry for beta amyloid. Eight autopsied brains with AD and seven with BD were compared with six control brains. The cortical microvessels in AD were frequently narrowed, and torn off, especially in close proximity to the senile plaques. The capillary densities in AD were significantly decreased as compared with the control brains. In contrast, there were no significant changes in the capillary densities and their morphologies in BD brains. Immunohistochemistry for beta amyloid revealed numerous deposits in the vascular wall and perivascular neuropil exclusively in AD brains. Cortical microvascular changes in AD and their absence in BD may indicate a role of beta amyloid for the microvessel pathology in AD. PMID- 17403575 TI - Are there any relationships among ankle proprioception acuity, pre-landing ankle muscle responses, and landing impact in man? AB - Proprioceptive input has been suggested to contribute to the pre-landing muscle responses associated with drop-landing, but its precise role has yet to be delineated. This study set out to examine the relationships among ankle proprioception, pre-landing muscle responses, and landing impact on drop-landing in healthy man. Fifteen healthy male basketball players aged 18 to 26 participated in this study. Passive ankle joint repositioning errors were used to examine ankle joint proprioception. Pre-landing EMG responses in the ankle muscles and the impact force on landing were recorded while the players performed self-initiated drops from a height of 30 cm. Results demonstrated that averaged ankle repositioning errors were significantly correlated with the co-contraction indexes between left tibialis anterior and medial gastrocnemius muscles (TA/MG CoI) (r=0.67, p=0.006), and showed a trend towards a relationship with the right TA/MG CoI (r=0.47, p=0.079). TA/MG CoI from both ankles were further related to the magnitude of the total impact force on landing (r=0.54 and 0.53, respectively; p<0.05). We concluded that male basketball players with less accurate ankle joint sense adopted greater co-contraction of ankle dorsiflexors and platarflexors, which was in turn associated with greater impact force at the moment of landing. PMID- 17403576 TI - Association of GST M1 null polymorphism with Parkinson's disease in a Chilean population with a strong Amerindian genetic component. AB - We have studied the association of a null mutation of Glutathione Transferase M1 (GST M1*0/0) with Parkinson's disease (MIM 168600) in a Chilean population with a strong Amerindian genetic component. We determined the genotype in 349 patients with idiopathic Parkinson's disease (174 female and 175 male; 66.84+/-10.7 years of age), and compared that to 611 controls (457 female and 254 male; 62+/-13.4 years of age). A significant association of the null mutation in GST M1 with Parkinson's disease was found (p=0.021), and the association was strongest in the earlier age range. An association of GSTM1*0/0 with Parkinson's disease supports the hypothesis that Glutathione Transferase M1 plays a role in protecting astrocytes against toxic dopamine oxidative metabolism, and most likely by preventing toxic one-electron reduction of aminochrome. PMID- 17403577 TI - Postural control after a strenuous treadmill exercise. AB - The effect of a strenuous treadmill exercise on body stability and the mechanisms associated with it have been studied with two different experimental protocols. The former investigation was based on stabilometric and metabolic measurements performed in basal condition and after a strenuous treadmill exercise whilst the latter dealt with the study of the early postural response to a 3s-bilateral soleus muscle vibration after the strenuous exercise. Our exercise protocol was able to induce an important generalized metabolic fatigue, as assessed by the obtained peak values in the measured metabolic parameters, and resulting in a short-lasting body destabilization. A linear relationship between sway path and oxygen uptake was found. Thus, the short duration of body instability could be likely due to the quite rapid recovery of oxygen uptake. Further, the fatigue induced body instability did not associate with changes in the early postural response to soleus muscle vibration. The present study cannot rule out the possibility that further central and/or peripheral mechanisms, influencing the postural control, may play a role in the fatigue-induced changes in body sway. PMID- 17403578 TI - Neurosensory changes in a human model of endothelin-1 induced pain: a behavioral study. AB - Although pain is a frequent feature in patients with cancer, its etiology is still poorly understood. In recent years, endothelin-1 (ET-1) has become a major target molecule in the etiology of cancer pain. In this randomised, double-blind study the effects of intradermal injection of ET-1 on spontaneous pain, temperature perception and sensation of punctate stimulation were evaluated. Thirty-five subjects were randomised to receive either placebo or one of four concentrations of ET-1 (ranging from 10(-10) to 10(-6)M). Besides assessment of spontaneous pain, three neurosensory testings were performed: (1) cold and warm sensation, (2) cold and heat pain, and (3) punctate stimulation using a von Frey monofilament. ET-1 produced a dose-dependent flare zone that was absent after placebo injection. Subjects reported a short-lasting spontaneous pain upon administration of the highest concentrations of ET-1. Injection of ET-1 induced a long-lasting and dose-dependent punctate hyperalgesia in an area around the injection site (secondary hyperalgesia). Thermal testing revealed a short period of hypoesthesia to non-noxious warm and cold stimuli after some doses of ET-1. In addition to the mechanical hyperalgesia, intradermal injection of ET-1 almost instantaneously induced a state of cold hyperalgesia outlasting the study period (120 min). No development of heat hyperalgesia was observed. The observed psychophysical characteristics of this new model of ET-1 induced nociception indicate its potential as a human experimental model for cancer pain. PMID- 17403579 TI - Theoretical analysis of effect of tip atomic species on apparent barrier height on Al(100) surface. AB - We have analyzed the effect of the tip atomic species on the tip-sample separation and the bias-voltage dependence of apparent barrier height (ABH) on an Al(100) surface using the boundary-matching scattering-state density functional method, which can be used to calculate electron states under applied bias voltages self-consistently within the density functional theory. We found that, from the dependence of the tip-sample separation, the difference between measurements with the two tip atomic species is larger in the ABH than in the maximum barrier height evaluated from the calculated potential profile. Furthermore, we found that the bias-polarity dependence of the ABH measured with the Na tip shows behavior opposite to that shown by the ABH measured with the Al tip. These results can be understood from the difference in the degree of lateral confinement of tunneling electrons in the tunneling barrier region between the two atomic species. PMID- 17403580 TI - Effects of 10 MeV electron irradiation at high temperature of a Ni-Mo-based Hastelloy. AB - A Hastelloy alloy was irradiated with 10 MeV electrons at 650 degrees C for 700 h to a total dose of 2 x 10(-3) displacements per atom (dpa). The microstructure of irradiated and non-irradiated specimens of this alloy were investigated by transmission electron microscopy (TEM). The non-irradiated specimens were analyzed by 3-D atom probe tomography (APT) in a local-electrode atom-probe (LEAP). TEM analysis before the irradiation detects small precipitates with a mean diameter of 22 nm, which are coherent with the FCC matrix. The number density of these precipitates is approximately 7 x 10(18) m(-3). Electron diffraction patterns from these precipitates exhibit superlattice reflections corresponding to the L1(2) ordered structure. The chemical composition of the precipitates, as measured by APT, is around 75 at% Ni with additions of Al, Ti and Mo. After electron irradiation, small precipitates with an irregular morphology are observed. The number density of these new precipitates about 10(20) m(-3) is greater than that of the L1(2) ordered precipitates before irradiation. The L1(2) superlattice reflections disappear completely, instead diffuse diffraction spots are observed at 1(1/2)0(FCC), which is attributed to compositional short-range order (SRO). The results are discussed with respect to the influence of the electron irradiation on the morphology and structure of the ordered precipitates. PMID- 17403581 TI - Atom probe specimen preparation with a dual beam SEM/FIB miller. AB - Dual beam scanning electron microscope/focused ion beam (SEM/FIB) methods complement electropolishing methods and enable specimens to be made from a wider range of materials. Several methods have been developed to fabricate specimens from different forms of materials, including thin ribbons, mechanically ground sheet and fine powders. In addition, FIB-based methods can be used in conjunction with electropolishing methods to improve the shape, surface finish and taper angle of specimens. Several lift-out (LO) methods have been developed for selecting specific microstructural features or other regions of interest such as phases, interfaces, grain boundaries, subsurface or implanted regions and interdendritic regions. These LO methods make use of an in situ nanomanipulator and platinum deposition to transfer and attach the lifted out volume to a post for final annular milling into a needle-shaped specimen. In order to improve the efficiency and to facilitate the LO procedure, some special specimen mounts that hold both the specimen and the support post at the appropriate working distance have been developed. PMID- 17403582 TI - Comparison of serological assays for the diagnosis of canine visceral leishmaniasis in animals presenting different clinical manifestations. AB - Three serological methods, indirect fluorescent immunoassay (IFI), enzyme-linked immunosorbent assay (ELISA) and direct agglutination test (DAT) that are commonly employed in the diagnosis of canine visceral leishmaniasis (CVL), have been assessed. A total of 234 domestic dogs, drawn from an area in the municipality of Belo Horizonte, Minas Gerais, Brazil, endemic for visceral leishmaniasis, were submitted to clinical and parasitological examinations and serological assay. Sera collected from confirmed non-infected dogs (n=20), and from dogs with other parasitic diseases including Trypanosoma cruzi (n=7), Leishmania braziliensis (n=5), Toxoplasma gondii (n=5) and Ehrlichia canis (n=3), were also included in the study. IFI presented a lower sensitivity (72%) than ELISA (95%), although the specificities of these assays were low (52 and 64%, respectively) and both exhibited cross-reactivity with sera from dogs infected with T. cruzi, L. braziliensis and E. canis. In contrast, DAT exhibited a high sensitivity (93%) and a high specificity (95%) and cross-reacted with only one serum sample derived from an E. canis-infected dog. The reproducibilities of the ELISA and DAT assays were excellent, whilst that of IFI was considered to be acceptable. The results produced by ELISA and DAT were in complete agreement, those between ELISA and IFI were at an acceptable level of agreement, whilst the concurrence between the IFI and DAT results were either acceptable or poor depending on the clinical conditions of the group of dogs examined. Since there is no readily accessible method for the diagnosis of CVL that offers 100% specificity and sensitivity, the choice of technique employed must depend on the aim of the investigation. PMID- 17403583 TI - Epidemiology of subclinical dairy cow nematode infections on five farms in England in 2002 and a comparison with results from 1978 to 1979. AB - A detailed study of the epidemiology of subclinical nematode infections in adult dairy cows was conducted on five well-managed English dairy farms during the 2002 grazing season. These same farms had also participated in a similar study in 1978 1979 and thus provided a unique opportunity to compare the epidemiological findings after an interval of over two decades. Common factors, including the prevalence of infection, faecal worm egg output, pasture larval count and nematode genera present, and estimated daily larval intake at pasture, were compared between the two studies. Subclinical roundworm burdens, as judged by faecal egg counts, were widespread in dairy cows in 2002, but the prevalence of animals with patent infections and the magnitude of the worm egg output were significantly lower than in 1978-1979. Both the prevalence of infection and mean faecal worm egg output exhibited a marked seasonal peak during the summer months in 2002. Pasture larval numbers were, however, nearly three times higher in 2002 with Ostertagia dominating the nematode genera to which cows were exposed at pasture in both surveys. Factors considered likely to account for differences in parasite epidemiology include the marked increase in herd productivity over the intervening period, the increased proportion of Holstein genetics with the consequent increase in milk yield and feed intake, changes in grassland management and increased stocking rate seen on the majority of farms. PMID- 17403584 TI - Differences between simultaneous and blocked training detected by a transfer test. AB - Secondary data analysis was used to compare responding early on a transfer test from rats previously trained simultaneously or successively on multiple temporal discriminations for the same number of trials [Guilhardi, P., Church, R.M., 2005 a. Dynamics of temporal discrimination. Learn. Behav., 33, 399-416]. Three fixed intervals (30, 60, and 120 s) were signaled by three stimuli (light, noise, and clicker). Twelve rats were trained with the three stimulus-interval pairs intermixed on each experimental session (simultaneous condition); 12 other rats were trained in successive blocks of 10 sessions on each pair (blocked condition). Then, all rats had a transfer test in which all three stimulus interval pairs were presented intermixed on each session. Rats in the simultaneous and blocked condition responded similarly during training, but differently during early stages of the transfer test. One possibility is that rats in the blocked condition were controlled by the previous interval, not by the current stimulus. These results challenge the usual assumptions from models of timing and conditioning that both simultaneous and blocked training produce learning of the associations between stimulus and interval in a multiple interval training task. PMID- 17403585 TI - Social leaning and stereotypy in horses. PMID- 17403586 TI - Direct effects of lead (Pb2+) on the relaxation of in vitro cultured rat aorta to acetylcholine. AB - Lead (Pb(2+)) exposure is related to increased blood pressure or hypertension of human or animals. Abnormal vascular relaxant responses of low level Pb(2+) exposed animals were reported by several studies. However, it is difficult to tell whether these effects were induced directly by Pb(2+) or not. In this study we hypothesized that Pb(2+) can directly affect the relaxation of vessels. Male Wistar rat aortae were removed and cultured in PMRI 1640 with 1 ppm Pb(2+) (4.8 microM lead acetate) for 0.5, 6, 12 and 24h, and then their responses to acetylcholine (ACh) and sodium nitroprusside (SNP) were examined. After incubated for 24h, the relaxation induced by ACh was significantly decreased in Pb(2+) exposed aortic rings. However, there was not significant difference in relaxation induced by SNP between Pb(2+) exposed and control group. The nitrite in the culture media of aortic rings cultured for 24h, measured with Griess method, was significantly decreased in the Pb(2+) exposed group. The expression of endothelial NOS (eNOS) and isoform NOS (iNOS) in the homogenate of aortic rings cultured for 24h was measured by Western blot. The expression of eNOS of the Pb(2+) exposed group was significantly upregulated compared with that of the control group. However, there was no significant difference in the expression of iNOS in control and Pb(2+) exposed group. In conclusion, Pb(2+) was able to directly affect the relaxation of rat aorta. This effect may have some relation with the lower level of NO in the media, though the expression of eNOS was upregulated. PMID- 17403587 TI - p-Phenylenediamine induced DNA damage in SV-40 immortalized human uroepithelial cells and expression of mutant p53 and COX-2 proteins. AB - p-Phenylenediamine (p-PD) is the main aromatic amine used in the formulation of hair dyes. Some epidemiologic studies have suggested that the use of p-PD-based hair dyes might be related to increased risk of human malignant tumors including bladder cancer and hematopoietic cancers. However, the toxicity and genotoxicity of p-PD on urothelial cells has not been reported yet. Therefore, we investigated the genotoxicity of p-PD on human urothelial cells and study its association with the expression of oncoproteins p53 and cyclooxygenase-2 (COX-2). Our results revealed that p-PD was able to induce DNA damage determined by Comet assay. In addition, our immunocytochemical and Western blotting results showed that p-PD induced overexpression of mutant p53 and COX-2 in a dose-dependent manner. The relationship between mutant p53 and COX-2 expression shows strong correlation. Furthermore, the accumulation of mutant p53 was linearly correlated with Comet scores. These results suggest that p-PD can induce DNA damage and accumulation of mutant p53 and COX-2 proteins; this may be one of the possible mechanisms that cause genotoxic carcinogenesis in the urothelial cells. PMID- 17403588 TI - Progesterone and the distribution of pituitary gonadotropin isoforms in cattle. AB - The objective of the present study was to determine the relative proportion of gonadotropin isoforms in bovine pituitary glands affected by progesterone. Twelve postpubertal heifers (Swiss-Zebu) were assigned to three groups (n=4): intact animals in the luteal phase of the estrous cycle (diestrus group); ovariectomized heifers with (OVXP) or without progesterone treatment (OVX). Prior to pituitary gland collection, a blood sample was taken from each animal to determine the circulating progesterone concentration. Pituitary protein extractions processed by chromatofocusing were eluted with a pH gradient ranging from 10.5 to 3.5. The LH and FSH eluent was grouped on the basis of the following three criteria: (1) as either a basic (pH>or=7.5), neutral (pH 7.4-6.5) and acid (pHor=10.5-3.5); (3) on the basis of distinct isoforms 12 peaks of which (A-L) were identified for LH and 11 (I-XI) for FSH. The analysis by range of pH and by pH of elution in the OVX and OVXP groups showed no difference in the LH and FSH isoform ratio, but diestrus cattle differs having a greater ratio (p<0.05) of basic LH isoforms (87.5+/-0.4%) and lesser ratio (p<0.05) of acid isoforms (5.4+/-0.7%). In the diestrus group, the ratio of acid FSH isoform increased (62.1+/-1.7%), while neutral isoforms decreased (5.7+/ 0.4%, P<0.05). The analysis by isoform type of LH revealed a greater proportion of isoforms C (pH 9.4) and E (pH 9.0) in the groups with circulating progesterone when compared to the OVX group. The heterogeneity of FSH was quantitatively similar in most isoforms in the three groups, with the exception of the predominant isoform (VIII, pH 4.9) that was more abundant in the diestrus group (p<0.05). These results indicate that progesterone with other gonad factors influence the pituitary glicosylation altering the relative proportions of gonadotropin isoforms. PMID- 17403589 TI - Analgesic activity of Ugni molinae (murtilla) in mice models of acute pain. AB - Leaf extracts of Ugni molinae Turcz. (Myrtaceae) are used in Chilean folk medicine as analgesic and anti-inflammatory. The antinociceptive effect of dichloromethane (DCM), ethyl acetate (EA) and methanol (ME) leaf extracts was assessed by intraperitoneal, oral and topical administration in writhing, tail flick, and tail formalin tests in mice. The extracts showed a dose-dependent antinociceptive activity in all the assays under different administration routes. The ED(50) values for the different tests for the DCM, EA, ME extract and reference drug (ibuprofen) were as follows. Writhing test in acetic acid (i.p. administration): 0.21, 0.37, 1.37 and 0.85mg/kg, respectively; tail flick test (oral administration): 199, 189, 120 and 45.9mg/kg. The EC(50) values for tail flick test were (topical administration): 2.0, 0.35, 1.4 and 8.2% (w/v), respectively; and the topical analgesic effects were (formalin assay) 75.5, 77.5, 31.6 and 76.5%, respectively. Ugni molinae extracts produce antinociception in chemical and thermal pain models through a mechanism partially linked to either lipooxygenase and/or cyclooxygenase via the arachidonic acid cascade and/or opioid receptors. Flavonoid glycosides and triterpenoids have been isolated from the plant and can be associated with the observed effect. Our results corroborate the analgesic effects of Ugni molinae, and justify its traditional use for treating pain. PMID- 17403590 TI - Cadaver dogs--a study on detection of contaminated carpet squares. AB - INTRODUCTION: Cadaver dogs are known as valuable forensic tools in crime scene investigations. Scientific research attempting to verify their value is largely lacking, specifically for scents associated with the early postmortem interval. The aim of our investigation was the comparative evaluation of the reliability, accuracy, and specificity of three cadaver dogs belonging to the Hamburg State Police in the detection of scents during the early postmortem interval. MATERIAL AND METHODS: Carpet squares were used as an odor transporting media after they had been contaminated with the scent of two recently deceased bodies (PMI<3h). The contamination occurred for 2 min as well as 10 min without any direct contact between the carpet and the corpse. Comparative searches by the dogs were performed over a time period of 65 days (10 min contamination) and 35 days (2 min contamination). RESULTS: The results of this study indicate that the well-trained cadaver dog is an outstanding tool for crime scene investigation displaying excellent sensitivity (75-100), specificity (91-100), and having a positive predictive value (90-100), negative predictive value (90-100) as well as accuracy (92-100). PMID- 17403591 TI - MR findings of ovarian tumors with hormonal activity, with emphasis on tumors other than sex cord-stromal tumors. AB - Sex cord-stromal tumors including granulosa cell tumor, thecoma, Sertoli stromal cell tumor and steroid cell tumor are noted for their hormonal activity. However, there are many kinds of ovarian tumors other than sex cord-stromal tumors and tumor-like conditions with endocrine manifestations. Cross-sectional imaging, especially MR, can provide precise features of ovarian tumors and uterine morphological change even in a clinically latent excess of estrogen. In this article, we demonstrate typical imaging findings of ovarian tumors with hormonal activity. We also shortly explain the mechanism of the virilization and hyperestrogenism caused by ovarian tumors and tumor-like conditions. PMID- 17403592 TI - Simultaneous kinetic determination of levodopa and carbidopa by H-point standard addition method. AB - The kinetic H-point standard addition method (HPSAM) was applied to the simultaneous determination of levodopa and carbidopa. The method was based on the difference in the rate of oxidation of these compounds with Cu(II)-neocuproine system and formation of Cu(I)-neocuproine complex at pH 5.5. The absorbance of the Cu(I)-neocuproine complex was monitored at 453 nm. Experimental conditions such as pH, reagent concentrations, ionic strength and temperature were optimized. Simultaneous determination of levodopa and carbidopa was performed in the range of 0.8-4 and 0.2-1.5 microg ml(-1), respectively. The proposed method was applied to the simultaneous determination of levodopa and carbidopa in pharmaceutical samples, and satisfactory results were obtained. PMID- 17403593 TI - Investigation on response of the metabolites in tricarboxylic acid cycle of Escherichi coli and Pseudomonas aeruginosa to antibiotic perturbation by capillary electrophoresis. AB - Metabolomics is a new branch of systems biology exerting its influence in many aspects. In order to appraise the effects of antibiotics on central carbon metabolism, a CE based method was set up. With this platform, we estimated the organic acid metabolite pools' fluctuation of Escherichia coli and Pseudomonas aeruginosa cultured under 11 different antibiotics' stimuli. Multivariate data analysis showed that different antibiotics had clustered distributions for each strain and could be easily distinguished. Genetic, metabolic and antibiotic mechanism differences could also be deduced by the aid of further correlation analysis. For P. aeruginosa, even synergy action amid antibiotics could be ascertained. PMID- 17403594 TI - High performance liquid chromatography for quantification of gatifloxacin in rat plasma following automated on-line solid phase extraction. AB - An automated system using on-line solid phase extraction and HPLC with fluorimetric detection was developed and validated for quantification of gatifloxacin in rat plasma. The extraction was carried out using C(18) cartridges (BondElut), with a high extraction yield. After washing, gatifloxacin was eluted from the cartridge with mobile phase onto a C(18) HPLC column. The mobile phase consisted of a mixture of phosphoric acid (2.5mM), methanol, acetonitrile and triethylamine (64.8:15:20:0.2, v/v/v/v, apparent pH(app.) 2.8). All samples and standard solutions were chromatographed at 28 degrees C. The method developed was selective and linear for drug concentrations ranging between 20 and 600 ng/ml. Gatifloxacin recovery ranged from 95.6 to 99.7%, and the limit of quantification was 20 ng/ml. The intra and inter-assay accuracy were up to 94.3%. The precision determined not exceed 5.8% of the CV. High extraction yield up to 95% was obtained. Drug stability in plasma was shown in freezer at -20 degrees C up to 1 month, after three freeze-thaw cycles and for 24h in the autosampler after processing. The assay has been successfully applied to measure gatifloxacin plasma concentrations in pharmacokinetic study in rats. PMID- 17403595 TI - Differential expression of PKC isoforms in developing zebrafish. AB - Protein kinase C isozymes are a biologically diverse group of enzymes known to be involved in a wide variety of cellular processes. They fall into three families (conventional, novel and atypical) depending upon their mode of activation. Several classes of zebrafish neurons have been shown to express PKCalpha during development, but the expression of other isoforms remains unknown. In this study we performed immunohistochemistry to determine if zebrafish express various isoforms of PKC. We used antibodies to test for the presence of enzymes that are thought to be preferentially expressed in the nervous system (PKCgamma, betaII, delta, epsilon, theta and zeta). Here, we show that PKCgamma, epsilon, theta and zeta are expressed in the zebrafish CNS. Anti-PKCgamma labels Rohon-Beard sensory neurons and Mauthner cells. PKCepsilon and zeta staining is widespread in the CNS, and PKCtheta and betaII are expressed in skeletal muscle, especially at intersegmental boundaries. Immunoblot experiments confirm the specificity of the antibodies in zebrafish and indicate that the fish isoforms of PKCgamma, betaII, epsilon and zeta are similar to the mammalian isoforms. Interestingly, PKCtheta appears to be similar to PKCthetaII, which, to date, has been found exclusively in mouse testis, but not in the mammalian CNS. Overall, our findings indicate that several different PKC isoforms are expressed in zebrafish, and that Rohon Beard, Mauthner cells and muscle fibers preferentially express some isoforms over others. PMID- 17403596 TI - 'I wash until it feels right' the phenomenology of stopping criteria in obsessive compulsive washing. AB - Recent elaborations of cognitive behavioral theory in OCD suggest that difficulties in deciding when to stop a compulsive action may be related to the use of counter-productive termination criteria by obsessional patients [Salkovskis, P. M. (1999). Understanding and treating obsessive-compulsive disorder. Behaviour Research and Therapy, 37, s29-s52]. Such criteria are characterized by their subjective nature, i.e. a primarily internal reference point (e.g. 'just right' feelings), and are conceptualized as the "top level" of a general strategy involving elevated evidence requirements. Thirty-eight obsessional washers, 41 obsessionals with other problems and 43 healthy controls were interviewed about and rated two situations varying in the degree of urgency to wash; they also washed their hands in a behavioral test. Washers reported using subjective criteria more frequently and rated them as more important for the termination of the washes than the other groups in questionnaire, interview and laboratory data. Both obsessional groups considered more criteria before stopping than the healthy controls, suggesting that using multiple criteria is a general strategy. The data are consistent with the predictions of the elaborated cognitive-behavioral model of OCD. They indicate that the use of subjective criteria and elevated evidence requirements is affected by the perceived significance of the situation in a similar way for obsessional and non obsessional individuals. PMID- 17403597 TI - An experimental study of the role of vulnerability related perceptions in spider fear: comparing an imaginal and in vivo encounter. AB - The effect of manipulating perceptions of the uncontrollability, unpredictability and dangerousness of a spider was assessed using both an imaginal and in vivo task involving an encounter with a spider. Participants were randomly assigned to one of eight conditions formed by the crossing of factors. Experimental manipulations of uncontrollability, unpredictability and dangerousness all had a significant effect on Task Related Spider Fear in the in vivo exposure task. Results indicated a greater effect on task related fear for in vivo exposure (R(2)=.258) compared to imaginal exposure (R(2)=.053). Perceptions of spiders as uncontrollable, unpredictable and dangerous accounted for much of the variance in spider fear beyond that accounted for by the experimental manipulations. The idea that perceptions of spiders as uncontrollable, unpredictable and dangerous are causally related to spider fear was supported with in vivo exposure being a stronger modality for fear modification than imaginal exposure. PMID- 17403598 TI - Computer-aided diagnosis in chest radiography. AB - We have developed computer-aided diagnosis (CAD) schemes for the detection of lung nodules, interstitial lung diseases, interval changes, and asymmetric opacities, and also for the differential diagnosis of lung nodules and interstitial lung diseases on chest radiographs. Observer performance studies indicate clearly that radiologists' diagnostic accuracy was improved significantly when radiologists used a computer output in their interpretations of chest radiographs. In addition, the automated recognition methods for the patient and the projection view by use of chest radiographs were useful for integrating the chest CAD schemes into the picture-archiving and communication system (PACS). PMID- 17403599 TI - A 43kD protein from the herb, Cajanus indicus L., protects against fluoride induced oxidative stress in mice erythrocytes. AB - Present study has been carried out to evaluate the protective and curative roles of a 43kD protein isolated from the herb Cajanus indicus L. on sodium fluoride (NaF) induced toxicity in mice erythrocytes. In the preventive study, mice were divided into five groups consisting of six in each for the experiments. Group I animals got water and used as normal controls, animals of groups II and IV were exposed to 600ppm fluoride in water for 7 days. Animals of group III were treated with the protein (2mg/kg body weight) intraperitoneally for 7 days followed by NaF treatment for next 7 days (600ppm). Animals of group V were treated with NaF (600ppm) followed by the protein treatment (2mg/kg body weight) for the same time as mentioned for group III. In the curative study, four groups of six mice were compared. Erythrocytes were isolated, and the antioxidant enzyme superoxide dismutase (SOD) as well as the levels of reduced and oxidized glutathione (GSH and GSSG), total thiols and lipid peroxidation end products were measured in those cells. There was a significant increase in lipid peroxidation along with a decrease in total thiols and SOD activity in the erythrocytes of NaF only and NaF bovine serum albumin treated animals. The 43kD protein treatment in animals either prior or post to fluoride administration normalized the levels of parameters measured and restored the SOD activity in mice erythrocytes. Data suggest that the 43kD protein possesses significant protective and curative activity against NaF induced oxidative stress in erythrocytes. PMID- 17403600 TI - Ion transport, membrane fluidity and haemoglobin conformation in erythrocyte from patients with cardiovascular diseases: Role of augmented plasma cholesterol. AB - Tissue hypoxia, which plays a key role in the development of renal and vascular complications of cardiovascular diseases (CVD), might be considered a consequence of vascular remodeling and/or attenuated oxygen (O(2)) delivery by erythrocytes. Using Raman spectroscopy (RS), we observed that erythrocytes from patients with CVD exhibit changes in the conformation of haemoglobin (Hb) haemoporphyrin (HP), reflecting its lower O(2) transport capacity. Hypertriglyceridemia and hypercholesterolemia are well-known hallmarks of CVD. This study examined the role of plasma lipids in the regulation of erythrocyte membrane viscosity, oxy-Hb content as well as Na(+)/H(+) exchange and Ca(2+)-ATPase, whose activities are altered in patients with CVD. HP conformation was assessed by RS of blood samples. Membrane fluidity was estimated at depths of 0.6-0.8 and 2.2nm by electron-paramagnetic resonance spectroscopy of erythrocytes loaded with spin labeled 5-doxylstearic acid and 16-doxylstearic acid, respectively. Ion-selective electrodes were employed for the study of H(+) and Ca(2+) fluxes. Both oxy-Hb content and erythrocyte membrane fluidity were decreased in essential hypertension and coronary artery disease patients and negatively correlated with plasma cholesterol but not triglyceride content. This observation allows us to assume that decreased oxy-Hb content in patients with CVD is caused by high plasma cholesterol via attenuation of erythrocyte membrane fluidity and its permeability to O(2). Plasma cholesterol level correlated positively and negatively with erythrocyte Na(+)/H(+) exchange and Ca(2+)-ATPase, respectively. However, in contrast to membrane fluidity, the impact of these ion transporters in oxy-Hb regulation under baseline conditions seems to be negligible. We propose that decreased oxy-Hb content contributes to the reduced O(2) tissue supply seen in patients with CVD. PMID- 17403601 TI - Unusual medium-chain polyunsaturated fatty acids from the snow alga Chloromonas brevispina. AB - The gas chromatography-mass spectrometry (GC-MS) method was used to identify unusual medium-chain polyunsaturated fatty acids (PUFAs) in the snow alga Chloromonas brevispina collected in 2006 from surface layers of a snow field with conspicuous green patches in Bohemian Forest (Czech Republic). PUFAs formed more than 75% total fatty acids. Among them, mass spectroscopy of picolinyl esters showed sizable proportions of medium-chain PUFA, e.g., 5,8,11-tetradecatrienoic and 6,9,12-pentadecatrienoic acids. The high relative content of PUFA indicates that PUFA are an important element ensuring cell survival. Our report appears to be the first to describe the presence of short- and medium-chain PUFAs in green psychrophilic algae of the genus Chloromonas. PMID- 17403602 TI - Vitreoscilla hemoglobin aids respiration under hypoxic conditions in its native host. AB - When Vitreoscilla were grown in medium containing 60mM sodium nitrite under both normal and limited aeration conditions, the levels of Vitreoscilla hemoglobin (VHb) were decreased by greater than 90%, while the levels of the terminal respiratory oxidase, cytochrome bo, were increased 350% under normal aeration and 7-23% under limited aeration. Cytochrome function, as measured by both NADH and ubiquinol oxidases for cells grown under both conditions, increased in parallel (by 150-222% and 8-56%, respectively, for the two activities). Nitrite in the medium inhibited Vitreoscilla growth at both normal and limited aeration. The inhibition of VHb at 60mM nitrite decreased whole cell respiration to the greatest degree in stationary phase for growth in limited aeration conditions, which was the most oxygen poor condition tested. These results are consistent with the originally proposed role for VHb, as an aid to respiration under hypoxic conditions. PMID- 17403603 TI - Feeding flaxseed oil but not secoisolariciresinol diglucoside results in higher bone mass in healthy rats and rats with kidney disease. AB - Flaxseed's oil and lignan, secoisolariciresinol diglucoside (SDG), are implicated in attainment of health and treatment of renal injury and osteoporosis. To test for these benefits, weanling Han:SPRD-cy rats (n=171) with or without kidney disease were randomized to diets made with either corn oil or flaxseed oil and with or without SDG for 12 weeks. In females, weight was lower with the SDG diet. In males fed flaxseed oil, lean mass was higher and fat % was lower. In both sexes, fat % was lower in diseased rats. Bone mineral content (BMC) and density were higher in rats fed flaxseed oil and lower in diseased rats, additionally; BMC was lower in SDG-supplemented females. The benefit of flaxseed oil on body composition is sex specific but the effect on bone mass is not. Lastly, reduced weight due to early rat kidney disease is not due to loss of lean body mass. PMID- 17403604 TI - A comparison of the neutron thermalization and reflection methods used for bulk hydrogen analysis. AB - The advantages and limitations of the slowing down of epithermal neutrons and the thermal neutron reflection methods were compared for bulk hydrogen analysis. In both cases, the same sample dimensions were used, i.e. 10cm diameter and 8cm height. It was found that the sensitivity of the slowing down method is higher by a factor of approximately 20 in average and the matrix effect can be neglected as compared to the neutron reflection technique. The simultaneous application of the two methods renders the determination of the H content and the C/H atomic ratio possible. The effect of spatial distribution of hydrogen in the matrix to be investigated on the number of reflected neutrons was also studied. PMID- 17403605 TI - Pulmonary sequestration with unusually large feeding vessel. PMID- 17403606 TI - Continuous veno-venous hemodiafiltration in children after cardiac surgery. AB - OBJECTIVE: Acute renal failure (ARF) is still a frequent complication following extensive cardiac surgery. Renal replacement therapy (RRT) modality preferences to treat critically ill children have shifted from peritoneal dialysis to continuous renal replacement therapy (CRRT), although the experience with the latter is still highly limited in the infants. METHODS: We describe our results with continuous veno-venous hemodiafiltration (CVVHDF) in 25 children (15 males, 10 females) who underwent CRRT from 2001 to 2006 and were retrospectively reviewed. RESULTS: We performed continuous veno-venous hemodiafiltration (CVHDF) using PRISMA (Hospal). The mean age at the onset of CRRT was 26 months (ranging from 7 days to 11.2 years) and the mean body weight was 14 kg. The mean duration of RRT was 67 h (8-243 h) with ultrafiltration rate 4.9 ml/(h kg); the mean filter "lifetime" was 31.5h. Anticoagulation was achieved with non-fractioned heparin infusion (21/25 cases) and enoxaparin (2/16). The mean creatinine concentrations at the beginning, 24, 48 and 72 h were as follows: 171, 100, 65 and 88 micromol/l. Of these 25 treated children, 19 died in the postoperative period (8 during CVVHDF). The mortality rate for the entire group was 76%. The main cause of death was cardiac failure and sepsis with multiorgan dysfunction (MODS). The main complication during CRRT was bleeding, transient hypothermia, thrombocytopenia and filter clotting which occurred in about one-third of the patients. CONCLUSIONS: We conclude that CVVHDF may be an alternative method of renal support for critically ill children after cardiac surgery in experienced centers, but a significant number of specific complications should be taken into account. PMID- 17403607 TI - Preservation of self: an overview of E3 ubiquitin ligases and T cell tolerance. AB - Until recently ubiquitination of a protein was thought to simply serve the mundane task of targeting a protein for proteasomal degradation. Accumulating evidence over the past decade has demonstrated the importance of ubiquitination in non-degradative functions including regulating cellular signaling, that highlight its role in human disease and thus potential development of novel therapeutics. Much has been written about ubiquitination in the immune system, in this review we will outline our current knowledge of ubiquitination with respect to T cell tolerance. Specifically, we will provide on overview of E3 ubiquitin ligases and their role in various states of CD4+ T cell tolerance: central and peripheral. PMID- 17403608 TI - Epilepsy and migraine in a patient with Urbach-Wiethe disease. AB - We report the clinical, neuroradiological, and molecular genetic findings in a patient with lipoid proteinosis or Urbach-Wiethe disease. Interestingly, in this patient epilepsy and migraine were the symptoms leading to the diagnosis of the disease, contrary to most patients in whom skin abnormalities are the first recognized symptoms. PMID- 17403609 TI - A post-streptococcal pathogenesis in children with tic disorders is suggested by a color Doppler echocardiographic study. AB - BACKGROUND: A post-streptococcal autoimmune mechanism, similar to that of rheumatic fever or Sydenham's chorea, has been hypothesized in some cases of neuropsychiatric disorder (tics and/or obsessive-compulsive disorders). A few studies on the involvement of other organs, outside the central nervous system, have been performed in these patients. AIM: To evaluate a possible post streptococcal pathogenesis in the children affected by tic disorders and showing sign of streptococcal exposure. METHODS: A case-control study was performed at the Outpatient Division of the Child Neurology and Psychiatry, and Paediatrics Departments of the University "La Sapienza" of Rome, from September 1, 2000, to February 28, 2005. Forty-eight subjects affected by tic disorder, aged 4-16 years, with signs of a recent or intercurrent exposure to streptococcal antigens, and 18 age-matched patients affected by tic disorder but without evidence of streptococcal exposure were examined by Color doppler echocardiography. RESULTS: The rate of echocardiographic abnormalities was significantly higher (p<0.001) in the patients with sign of streptococcal exposure. In 28 out of 48 patients (58.3%), the color Doppler echocardiography showed abnormalities: 26 patients (54,3%) had a mitral regurgitation, 1 (2%) a mitral valve prolapse and finally 1 (2%) showed a kinking of the anterior mitral valve leaflet. In the control group, four children (22.2%) showed a mitral regurgitation. All of these abnormalities were not hemodynamically significant, and in many cases decreased with time. CONCLUSIONS: The higher rate of echocardiographic abnormalities observed in patients with tic disorder and exposure to group A beta-haemolytic streptococcal antigens, together with their decrease with time, suggest a post-streptococcal pathogenesis. PMID- 17403610 TI - Motor coordination in children with congenital strabismus: effects of late surgery. AB - BACKGROUND: Strabismus is one of the most common visual disorders in infancy. While there is a great attention on the effects of the timing of surgery as to the development of binocular vision, little is known about the possible influence of congenital strabismus on perceptual-motor and more generally, on neuromotor development. AIMS: Aim of this study was to investigate perceptual-motor and motor coordination abilities of 19 children with essential congenital esotropia who underwent a late surgery (after 4 years), compared to 23 age-matched controls. METHODS: Children were tested using the Movement Assessment Battery for Children (Movement ABC) that were performed both 1-week before surgery (T1) and about 3 months (+/-2 weeks) after surgery (T2). RESULTS AND CONCLUSIONS: At T1, abnormal or borderline results were found in more than half of the children with strabismus, as opposed to only about 17% of the controls. At T2 none of the children showed abnormal Movement ABC total scores and there was no difference in global scores between the study and the control group. The two groups also did not show any significant difference in individual items of the movement ABC with the exception of those assessing ball skills. Our results suggest that surgical correction of strabismus, even when performed after the 4th year of life, appears to be effective in improving perceptual-motor and motor function. PMID- 17403611 TI - A low-cost telemetry system suitable for measuring mouse biopotentials. AB - The ability to generate specific genetic mutations in mice is a powerful tool to study normal and pathophysiological function. In order to determine the effects of a mutation, measurement of physiological variables, such as biopotentials, is often necessary. However, such measurements can be particularly challenging to obtain from an awake, unrestrained mouse. The goal of this study was to design and implement a telemetry system suitable for recording biopotentials from a mouse. A battery-powered system was fabricated from commercially available electronic components mounted on a small circuit board. The frequency response of the system was measured over a range of frequencies and found suitable for recording biopotentials in mice and larger animals. We affixed the circuit board externally to a mouse and connected surface electrodes to measure electrocardiograms (ECG). The size and weight of the board did not disturb normal behavior over 30-60 min. Recorded ECGs had easily identifiable components relevant to physiological parameters and had a similar frequency spectrum compared to recordings obtained from a commercially available measurement system. In conclusion, the telemetry system was low-cost due to the availability of the components, straightforward to implement, and provided biopotential recordings suitable for measuring physiological parameters in an awake mouse. PMID- 17403612 TI - TRAIL signalling: decisions between life and death. AB - The TNF-related apoptosis-inducing ligand, TRAIL, has been shown to selectively kill tumour cells. This property has made TRAIL and agonistic antibodies against its death inducing receptors (TRAIL-R1 and TRAIL-R2) to some of the most promising novel biotherapeutic agents for cancer therapy. Here we review the signalling pathways initiated by the apoptosis- as well as the non-apoptosis inducing receptors, TRAIL-R3 and TRAIL-R4. The TRAIL "death-inducing signalling complex" (DISC) transmits the apoptotic signal. DISC formation leads to activation of a protease cascade, finally resulting in cell death. The TRAIL death receptor-mediated "extrinsic" pathway and the "intrinsic" pathway, which is controlled by the interaction of members of the Bcl-2 family, interact with each other in the decision about life or death of a cell. Apoptotic and non-apoptotic signalling is influenced by the NF-kappaB, PKB/Akt and the MAPK signalling pathways. In this review we intend to summarise the most important findings on the TRAIL signalling network and the interplay in the decisions between life and death of a tumor cell. PMID- 17403613 TI - N-terminal pro brain natriuretic peptide to predict prognosis in dilated cardiomyopathy with sinus rhythm. AB - AIMS: To assess the value of plasma NT proBNP levels for predicting adverse outcomes in patients with dilated cardiomyopathy (DCM). METHODS: Seventy-eight patients with DCM (EF <40%) with sinus rhythm were enrolled. All patients had undergone echocardiographic examination, coronary angiography, and cardiac catheterisation. Blood samples for plasma NT proBNP levels were taken at rest following echocardiographic examination. Patients were followed up for 660+/-270 days for clinical endpoints defined as; death from worsening heart failure, sudden cardiac death and heart transplantation (Tx). RESULTS: Clinical end points were observed in 19 patients (5 Tx, 4 sudden cardiac death, 10 death from worsening heart failure). Variables associated with an increased hazard of clinical endpoints in univariate analysis were log NT proBNP, age, NYHA functional class, left ventricle ejection fraction, mitral valve effective regurgitation orifice area, and E wave deceleration time. The plasma level of NT proBNP (Hazard ratio=2.5 [95% CI: 1.3-4.7], p=0.0024) and age (hazard ratio=0.94 [95% CI: 0.90-0.98], p=0.0005) were the independent variables associated with an increased risk of clinical endpoints. NT proBNP plasma level >4500 pg/ml detected patients with clinical endpoints with a sensitivity, and specificity of 72%, 80%, respectively. The event free survival was found to be significantly lower in patients with NT proBNP levels >4500 pg/ml. CONCLUSION: NT proBNP seems to be a strong predictor of adverse outcomes in patients with DCM with sinus rhythm and may be used as a reliable biological marker in risk stratification. PMID- 17403614 TI - Duration, combination and timing: the signal integration model of dendritic cell activation. AB - The activation of resting dendritic cells (DCs) is a crucial step in the initiation of adaptive immunity because it links peripheral events initiated by the encounter with pathogens to the activation and expansion of antigen-specific T lymphocytes in secondary lymphoid organs. It is well recognized that a wide variety of microbial products and endogenous signals can trigger DC activation, and that different DC subsets are specialized in inducing different classes of immune responses. In this review, we will focus on how different aspects of DC maturation are regulated not only by the nature of the DC maturation stimuli, but also by their duration, combination and timing, and provide an overview of how different modes of DC activation can affect T cell responses. PMID- 17403615 TI - When killers become helpers. AB - Since their initial characterization by Kiessling over 35 years ago, natural killer (NK) cells continue to constitute an area of intensive discovery in the immunology field. Although most of the research efforts concentrated on characterizing the role of NK cells in tumor prevention and fighting infection through the killing of dangerous cells, several recent findings highlight unexpected non-cytolytic functions of human and mouse NK cells. Such functions include promoting placental tissue development, antigen presentation and stimulation of T cells, priming of macrophages and dendritic cells, reducing transplant tissue rejection and several others. PMID- 17403616 TI - Integrating noninvasive molecular imaging into molecular medicine: an evolving paradigm. AB - Molecular imaging is a rapidly emerging field, providing noninvasive visual quantitative representations of fundamental biological processes in intact living subjects. Fundamental biomedical research stands to benefit considerably from advances in molecular imaging, with improved molecular target selection, probe development and imaging instrumentation. The noninvasiveness of molecular imaging technologies will also provide benefit through improved patient care. Molecular imaging endpoints can be quantified, and therefore are particularly useful for translational research. Integration of the two disciplines of molecular imaging and molecular medicine, combined with systems-biology approaches to understanding disease complexity, promises to provide predictive, preventative and personalized medicine that will transform healthcare. PMID- 17403617 TI - Common XPD (ERCC2) polymorphisms have no measurable effect on nucleotide excision repair and basal transcription. AB - The xeroderma pigmentosum group D (XPD/ERCC2), a subunit of TFIIH, plays a critical role in nucleotide excision repair (NER) and basal transcription. There are hot spots of single nucleotide polymorphism (SNP) within the XPD gene sequence that have been incriminated in the pathophysiology of human cancers, possibly by altering the capacity of the cells for removing DNA damage induced by chemical adducts and UV radiation. A controversy persists on the role of these SNPs and this question has not been approached with appropriate biochemical tests. Thus, we sought to quantify in vitro, the effect of codon variants 201 (p.H201Y), 312 (p.D312N), 751 (p.K751Q) of XPD as well as the double XPD variant (p.D312N/p.K751Q) on NER and basal transcription. We used the baculovirus expression system to reconstitute recombinant TFIIH complexes in which the XPD variants were introduced and we analyzed their specific transcription and NER activities. Experimentally, variations in NER capacity and basal transcription activation of the four variants were not detectable in vitro. Structural analyses of XPD revealed that these single nucleotide polymorphisms sites were located outside the main catalytic domains. Altogether, evolutionary data, structural analyses and biochemical investigations strongly suggest that all XPD variants are comparable regarding the main properties of XPD and TFIIH. PMID- 17403618 TI - Translesion synthesis in Escherichia coli: lessons from the NarI mutation hot spot. AB - Duplication of DNA containing damaged bases is a challenge to DNA polymerases that normally replicate with high speed, high accuracy and high processivity undamaged templates only. When a replicative DNA polymerase encounters a chemically altered base that it is unable to copy, a process called translesion synthesis (TLS) takes place during which the replicative polymerase is transiently replaced by a so-called specialized or lesion bypass polymerase. In addition to the central players that are the replicative and translesion DNA polymerases, TLS pathways involve accessory factors such as the general replication processivity factor (i.e. the beta-clamp in prokaryotes and PCNA in eukaryotes). In Escherichia coli, besides the beta-clamp, RecA plays a fundamental role as a co-factor of Pol V the major bypass polymerase in this organism. An integrated view of TLS pathways necessarily requires both genetic and biochemical studies. In this review we will attempt to summarize the insights into TLS gained over the last 25 years by studying a frameshift mutation hot spot, the NarI site. This site was initially discovered by serendipity when establishing a forward mutation spectrum induced by a chemical hepatocarcinogen, N-2-acetylaminofluorene (AAF). Indeed, this chemical carcinogen covalently binds to DNA forming adducts with guanine residues. When bound to G* in the NarI site, 5'-GGCG*CC-, AAF induces the loss of the G*pC dinucleotide at a frequency that is approximately 10(7)-fold higher than the spontaneous frequency. In vivo studies showed that the NarI mutation hot spot is neither restricted to the NarI sequence itself, nor to the carcinogen AAF. Instead, the hot spot requires a sequence containing at least two GpC repeats and any of a family of aromatic amides and nitro aromatic compounds that form a large class of human carcinogens. Genetic analysis initially revealed that the NarI frameshift pathway is SOS dependent but umuDC (i.e. Pol V) independent. More recently, DNA Pol II was identified as the enzyme responsible of this frameshift pathway. Concurrently the AAF adduct in the NarI site can be bypassed in an error-free way by Pol V. The NarI site thus offers a unique possibility to study the interplay between two specialized DNA polymerases, Pol II and Pol V, that can both extend replication intermediates formed when the replicative Pol III dissociates in the vicinity of the damage. Full reconstitution of the two pathways led us to highlight a key feature for TLS pathways, namely that it is critical the specialized DNA polymerase synthesizes, during the course of a single binding event, a patch of DNA synthesis (TLS patch) that is long enough as to "hide the lesion induced distortion" from the proofreading activity upon reloading of the replicative DNA polymerase (or any exonuclease that may get access to the primer when the specialized DNA polymerase detaches). The beta-clamp, to which all DNA polymerases bind, plays a critical role in allowing the specialized DNA polymerases to synthesize TLS patches that are long enough to resist such "external proofreading" activities. PMID- 17403619 TI - High throughput quantification of phytoestrogens in human urine and serum using liquid chromatography/tandem mass spectrometry (LC-MS/MS). AB - Phytoestrogens are currently the subject of intense study owing to their potential protective effects against a number of complex diseases. However, in order to investigate the interactions between phytoestrogens and disease state effectively, it is necessary to have analytical methods which are sensitive, reproducible, and require low sample volumes. We report an assay for three isoflavones (daidzein, genistein, and glycitein), two metabolites of daidzein (equol and O-desmethylangolensin), three lignans (secoisolariciresinol, enterodiol, and enterolactone), and one flavanone (naringenin) in human urine and serum. A high throughput of samples has been achieved via the use of 96-well plate sample extraction and liquid chromatography/tandem mass spectrometry (LC MS/MS) analysis incorporating column switching, thus making the assay suitable for use on large sample numbers, such as those found in epidemiological studies. The robustness of the assay was proven via the comparison of data generated on two different LC-MS/MS systems, with and without column switching. PMID- 17403620 TI - Simultaneous determination of 13 amphetamine related drugs in human whole blood using an enhanced polymer column and gas chromatography-mass spectrometry. AB - Metamphetamine (MA) is one of the most frequently encountered abused drugs in Japan and the Triage immunoassay kit is often used to screen for this drug. However, immunoassay screening also gives positive results with other structurally related compounds, such as 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), p-methoxyamphetamine (PMA), an ephedrine metabolite and beta-phenethylamine (PEA). Therefore, it is important to develop a simple and reliable method which can determine these drugs simultaneously. This paper describes a simple method for simultaneous identification and quantification of 13 amphetamine related drugs in human whole blood. The method consists of a solid phase extraction using a new polar-enhanced Focus column followed by acetylation and gas chromatography-mass spectrometry in the scan mode. Tetradeuterated MA and trideuterated methylephedrine (ME) were used as internal standards. As the Focus column required only simple extraction steps and provided a clean extract, identification of each drug was feasible even at low concentrations. The calibration curves were linear over the concentration range from 50 to 5000 ng/ml for all drugs with correlation coefficients that exceeded 0.99. The lower limits of detection of the drugs were 5-50 ng/ml. The absolute recoveries for the drugs were 65-95% and 64-89% at concentrations of 100 and 1000 ng/ml, respectively. Accuracy and precision data were satisfactory when using 2 internal standards. The applicability of the assay was proven by the analysis of blood samples in forensic cases. This method should be most useful for confirmation of positive immunoassay results for amphetamines and related drugs. PMID- 17403621 TI - Changes in urinary level and configuration ratio of D-lactic acid in patients with short bowel syndrome. AB - The present study showed that the D-lactic acid configuration ratio in the urine rose earlier than that in blood or the urinary or blood D-lactic acid levels upon disease onset, and that the D-lactic acid measurement in urine is more sensitive and useful than that in blood. As this result, a prediction of a D-lactic acidosis may be possible. To simplify the procedure for detecting D-lactic acid, we first showed a correlation between the D-lactic acid configuration ratio in urine and blood, indicating urine could be used. To separate the optical isomers of lactic acid, we simplified our previous procedure. For chiral recognition, we chose O-acetyl-(-)-menthylation and analyzed the samples under GC/MS by capillary gas chromatography on a DB-5 MS column. This procedure is less sensitive than the former method, but it is faster and simpler, requiring only one derivatization step. This method may be useful for predicting D-lactic acidosis in patients with short bowel syndrome. PMID- 17403622 TI - Autoimmune phenomena as prognostic markers of response to adjuvant interferon therapy for melanoma. PMID- 17403623 TI - Me, onco-robot. PMID- 17403624 TI - Molecular pathology in sarcomas. AB - Bone and soft tissue sarcomas are an infrequent group of tumours. Their prevalence is 4 in 100,000 people/year, making the disease quite rare. Some of these tumours, such as synovial sarcoma, Ewing tumour and osteosarcoma, are more usual in adolescents or in young adults; there are, though, some neoplasias such as leiomyosarcoma or liposarcoma that are more frequent in patients over 55 years. There are more than a hundred different types of sarcomas from the histological point of view. This is the main limitation at the time of finding major clinic essays on patients with specific types of sarcomas. From the molecular point of view, these neoplasias are grouped into two main types: (a) sarcomas showing specific genetic alterations and relatively simple karyotypes, and translocations which originate gene fusions (e.g., EWS-FLI1 in Ewing tumour); or specific genetic mutations (e.g., c-kit in the gastrointestinal stromal tumour), and (b) sarcomas showing unspecific gene alterations and very complex karyotypes, and very numerous gains and losses. This review describes diverse types of molecular alterations as well, their utility in the clinical domain, as well as implications for the pathologist in translational research in sarcomas. PMID- 17403625 TI - Telomerase and telomere dynamics in ageing and cancer: current status and future directions. AB - This review will focus on the clinical utilities of telomerase for human cancer diagnosis and prognosis. Much attention has been focused on control of telomerase activity in early and late stage tumours. Telomerase stabilisation may be required for cells to escape replicative senescence and to proliferate indefinitely. Because of a very strong association between telomerase and malignancy, both clinicians and pathologists expect this molecule to be a useful diagnostic and prognostic marker and a new therapeutic target. These data have greatly inspired the development of various strategies to target telomere and telomerase for cancer therapy. Finally, evidence is now emerging that G quadruplex ligands produce rapid senescence and selective cell death. A summary of recent experimental works with new small molecules as potential inhibitors of telomerase is presented. PMID- 17403626 TI - microRNAs and cancer: role in tumorigenesis, patient classification and therapy. AB - MicroRNAs (miRNAs) are small non-coding RNAs that downregulate gene expression during various crucial cell processes such as apoptosis, differentiation and development. Recent work supports a role for miRNAs in the initiation and progression of human malignancies. Moreover, large high-throughput studies in patients revealed that miRNA profiling has the potential to classify tumours and predict patient outcome with high accuracy. Functional studies, some of which involve animal models, indicate that miRNAs act as tumour suppressors and oncogenes. This review examines the role of miRNAs in the pathogenesis of cancer as well as miRNA-profiling studies performed in human malignancies. Implications of these findings for the diagnosis and treatment of cancer patients are also discussed. PMID- 17403627 TI - Venous thromboembolic disease in cancer. Optimisation of the use of antithrombotic agents. AB - Venous thromoboembolism is one of the most common complications in cancer patients and may have serious consequences. At present, most clinical oncologists report using thromboprophylaxis in less than 5% of patients. One of the possible reasons for this limited use is the lack of oncology specific guidelines. In effect, while there are excellent guidelines for optimising the use of antithrombotic agents to prevent and treat thromboembolism, they must be adapted to the concrete context of cancer patients. The present review explores how different situations affect cancer patients and their risk of developing venous thromboembolism (VTE), and evaluates the situations in which antithrombotic agents should be administered to treat and prevent VTE. PMID- 17403628 TI - Clinical outcome in patients with intramedullary spinal cord metastases from lung cancer. AB - Intramedullary spinal cord metastases (ISCM) are uncommon and present with rapidly progressing neurological deficits. The objective of this study was to determine the rate, duration of neurological response and survival after radiation therapy. We have retrospectively reviewed the clinical outcome of six cases with a diagnosis of ISCM from primary lung cancer, non-small cell (NSCLC) (n=3) and small cell (SCLC) (n=3). Total radiation dose ranged from 27 Gy/5 fr to 40 Gy/20 fr. Ambulation was preserved in 3 patients and partially recovered in one. Five out of the six patients (83%) showed improvement in neurological signs/symptoms with a mean duration of 17.2 days (max: 40 days; min: 6 days). Median survival time was 5 months (confidence interval (CI) 95%: 0-12) for NSCLC and 5 months (CI 95%: 4-6) for SCLC. Although radiation response rate is high, the interval free of neurological progression is very short. A therapeutic approach should be considered for each individual. PMID- 17403629 TI - Weekly docetaxel with concomitant radiotherapy in patients with inoperable oesophageal cancer. AB - INTRODUCTION: To evaluate the efficacy and tolerability of weekly docetaxel concurrent with radiotherapy in inoperable oesophageal cancer patients. MATERIAL AND METHODS: Thirty-four oesophageal cancer patients with co-morbid medical conditions, locally advanced tumours (T4) or advanced age (older than 75 years) received docetaxel (20 mg/m2 weekly) plus concurrent radiotherapy (2 Gy daily, to a total dose of 66 Gy). Twenty-two patients (64%) were stage III, 19 of whom had T4 tumours. RESULTS: Twenty-seven patients (79%) completed the planned chemoradiotherapy treatment. Nine patients (26%) achieved a complete response and 8 (24%) achieved a partial response, for an overall response rate of 50%. Median survival was 6 months, and 1-year survival was 35%. Patients with T4 tumours had significantly shorter survival than other patients: 5 months for T4 tumours vs. 11 months for T1-3 (p=0.04). Grade 3-4 oesophagitis occurred in 6 patients (17%). There were two treatment-related deaths due to radiation pneumonitis. CONCLUSIONS: Docetaxel plus concurrent radiotherapy is active in poor-prognosis oesophageal cancer patients, with a lower incidence of severe oesophagitis than with cisplatin-based chemoradiotherapy regimens. This schedule can be considered, especially in patients with non-T4 tumours who are not candidates for oesophageal resection. PMID- 17403630 TI - Preoperative radio-chemotherapy (RT-CT) in rectal cancer. Prospective study with postoperative RT-CT control group. AB - INTRODUCTION: Between 1996 and 2000, the colorectal tumour committee of the Hospital Universitario de Bellvitge and the Institut Catala d'Oncologia, Hospitalet, carried out a non-randomised prospective study of pre-op radio chemotherapy (RT-CT) in locally advanced rectal tumours. We herein present the results. On the other hand, and at the same time, patients operated on for locally advanced rectal cancer were admitted and treated by RT-CT during the postoperative process, according to our standard protocol. Results for both series are compared. MATERIAL AND METHODS: The preoperative RT-CT group included 94 patients. They received radiotherapy (RT), 45 Gy on posterior pelvis, and simultaneously, 5-fluorouracil (5FU) by continuous infusion (300 mg/m2/day, 5 days weekly during RT). Surgical intervention was scheduled 6-8 weeks after preoperative treatment; after surgery they received 5FU (425 mg/m2/day) and leucovorin (20 mg/m2/day) bolus, 5 days weekly; 4 cycles at four-week intervals. 237 patients who had been previously operated on and who had been staged as T3-T4 and/or N+, M0 were admitted to our centre during the same time period and received postoperative RT-CT. RESULTS: The preoperative treatment group showed a complete and global response rate to RT-CT in 17% and 68% of cases, respectively. Anal sphincter was preserved in 38.5% of patients exhibiting low rectal tumours (inferior limit of tumour at 6 cm or less from the anal margin). Overall and disease-free survival at 5 years was distinct, showing statistical significance, according to the response obtained through preop treatment; it was better in responsive patients (overall survival: 87% in complete remissions, 75% in partial remissions, 48% in stable disease, and mean survival was 0.84 years for patients who evolved, p<0.05; disease-free survival was: 93% in complete remission, 76% partial remission, 39% in stable disease, p=0.001). We did not see any difference with regard to overall survival, disease-free survival or local control at the time of comparing either pre- or postoperative groups. There were, however, differences with regard to late toxicity; they showed less toxicity when RT-CT was administered preoperatively; no case of radiation enteritis that required surgery was seen in this group, whereas in the postoperative RT-CT it was 4.2%, p=0.022. CONCLUSIONS: Preoperative treatment of locally advanced rectal cancer is recommended, for it yields a high level of response to treatment; it allows preservation surgery of the anal sphincter in one third of patients showing low rectal tumours. There is also a clear diminution of late toxicity with pre-op treatment. On the other hand, response to pre-op treatment selects patients with a better prognosis. PMID- 17403631 TI - Aromatase inhibitors and male breast cancer. AB - The majority of breast cancers in male patients are hormone receptor positive. Tamoxifen has proven to be successful in both adjuvant and metastatic settings and remains the standard of care. Given the improved outcomes in female patients with aromatase inhibitors (AI), these drugs have become a potential therapeutic tool for male patients. Preliminary data show effective suppression of oestradiol levels in males treated with AI and some reports have demonstrated objective responses. Here we report a case of a male patient with metastatic breast cancer treated with letrozole who achieved clinical response associated with a decrease in blood oestradiol levels. PMID- 17403632 TI - Simultaneous presentation of oesophageal and renal cancer. AB - Oesophageal cancer has been documented to be often associated with other primary tumours. However, concurrent oesophageal and renal cell carcinoma is extremely uncommon. We report a case of synchronous oesophageal and kidney cancer that was successfully treated at our hospital by a one-stage surgical procedure. The patient is doing well and without recurrence 54 months after the operation. PMID- 17403633 TI - Superior vena cava syndrome with central venous catheter for chemotherapy treated successfully with fibrinolysis. AB - Recently, there has been an increase in the number of cases of superior vena cava (SVC) syndrome associated with chronic indwelling central venous catheters. Fibrinolytic therapy and endovascular treatment are currently achieving good results. We present a case history of a patient with SVC with a catheter used for chemotherapy, which was successfully treated with catheter-directed (intraclot) infusion thrombolytic therapy with urokinase. PMID- 17403634 TI - Will Rogers phenomenon in small-cell lung cancer. PMID- 17403635 TI - [Change in the cholesterol metabolism associated with the combined inhibition of synthesis and absorption]. AB - Lowering lipid levels in the cardiovascular prevention we confine ourselves to measure the cholesterol level and care less for the background effects. Namely blood cholesterol level beyond the amount consumed with the diet highly depends on balance of intestinal absorption/secretion and synthesis. Studying the rate of absorption and synthesis has come only recently into the foreground of interest. Many observations proved that using even the strongest cholesterol lowering drug beyond reducing the synthesis in the liver - may be associated with an up to 50 percent increase of the intestinal cholesterol absorption. When studying the effectiveness of statins in everyday practice we measure only the decrease of serum cholesterol level as the final result, and do not examine the changes in the synthesis and absorption. The amount of cholesterol synthesized or absorbed can be determined in an indirect way by measuring that of the non-cholesterol sterols (phytosterols). The absorption markers are campesterol, sitosterol, avenasterol as well as cholestanol. The biosynthesis of cholesterol correlates with the level of lathosterol, cholestanol, desmostenol. In practice the concentration of lathosterol or lathosterol/cholesterol can be considered the marker of synthesis and the campesterol or campesterol/cholesterol ratio the marker of absorption. So recent study results show that while inhibiting the cholesterol synthesis with statin the cholesterol absorption increases and the absorption inhibitor ezetimibe is associated with boost of synthesis. The increase in absorption caused by statins can be reduced or prevented by combining with ezetimibe. These data confirm that combination of statin and ezetimibe, inhibiting simultaneously both the synthesis and absorption provides the most effective cholesterol-level lowering with the least side-effects. PMID- 17403636 TI - [Arteriogenesis: a new strategy of therapeutic intervention in chronic arterial disorders. Cellular mechanism and experimental models]. AB - The term arteriogenesis became clarified only some years ago. This endogenous process is a natural compensation mechanism against stenosis or arterial occlusion-induced tissue hypoperfusion via improvement of blood distribution in the pre-existent collateral arteries. The main chronic artery disorders like coronary heart disease, peripheral artery disease and cerebrovascular disease were extensively studied for angiogenesis and arteriogenesis during the last decade. The in vivo animal experiments and the ex vivo analysis of the cellular and molecular mechanisms behind collateral artery development revealed the crucial role of circulating monocytes, endothelial and smooth muscle cells in the remodelling of collateral blood vessels. The adaptive arteriogenesis in the heart, brain and periphery can be stimulated by different chemokines and growth factors. The therapeutic application of these substances resulted in promising data in pre-clinical animal models, i.e. improved collateral conductance, extended neo-vascularization in the collateral dependent tissue regions, decreased infarct area after hemodynamic stroke and better functional parameters in myocardial ischemia. The questions that have to be addressed during the design of human investigations are the optimal delivery approach, the appropriate dosage, timing and the durability of the follow up. The present review tries to give an overview about the main points of the mechanism and the most important experimental data concerning spontaneous and stimulated collateral artery growth, this new and promising therapeutic approach for chronic artery diseases. PMID- 17403637 TI - [Interdisciplinary questions of stroke therapy]. AB - Numerous multicenter, randomized, preventive and therapeutic stroke trials have been initiated or already finished in the last years. Due to some positive results the stroke societies have elaborated the new guidelines for stroke prevention and therapy, based on the results of evidence based medicine. Unfortunately, in daily practice, the neurologist, vascular and cardiac surgeon, cardiologist or internist have to answer many interdisciplinary questions of stroke prevention or therapy. Because of the lack of large, international trials, the author summarizes the most important publications and his personal experience regarding the interdisciplinary questions (stroke in hospital, endarterectomy and CABG, stent or endarterectomy, complications after CEA) of stroke care. PMID- 17403638 TI - [Significance of dehydroepiandrosterone and dehydroepiandrosterone sulfate in different diseases]. AB - Dehydroepiandrosterone and dehydroepiandrosterone-sulfate are precursors of androgens and estrogens, support the gonadal sexual steroid production. The levels of dehydroepiandrosterone and dehydroepiandrosterone-sulfate are maximal between the ages of 20 and 30 years, then start a decline of 2% per year, leaving a residual of 10-20% of the peak production by the eight decade of life. The age associated decrease may lead to osteoporosis, deterioration of lipid-metabolism, cardiovascular diseases and second type of diabetes mellitus. Decreased levels were found in autoimmune diseases and in sexual dysfunction, too. Intracrinology describes the formation of active hormones which exert their action in the same cells where synthesis took place without release into the pericellular compartment. The high local androgen and estrogen concentration may be important in the pathomechanism of hirsutism, acne, seborrhea, breast and prostate cancer. Administration of dehydroepiandrosterone resulted in a reduction of postmenopausal osteoporosis, also the decreased symptoms in systemic lupus erythematosis, psychiatric diseases and sexual disfunction. The authors summarize the metabolism of dehydroepiandrosterone and dehydroepiandrosterone-sulfate and their role in different diseases. PMID- 17403639 TI - [On pathophysiology of stress]. PMID- 17403641 TI - [Hungarian brains endangered. Interview with Szilveszter E. Vizi, President of the Hungarian Academy of Sciences]. PMID- 17403642 TI - Levels of processing during non-conscious perception: a critical review of visual masking. AB - Understanding the extent and limits of non-conscious processing is an important step on the road to a thorough understanding of the cognitive and cerebral correlates of conscious perception. In this article, we present a critical review of research on subliminal perception during masking and other related experimental conditions. Although initially controversial, the possibility that a broad variety of processes can be activated by a non-reportable stimulus is now well established. Behavioural findings of subliminal priming indicate that a masked word or digit can have an influence on perceptual, lexical and semantic levels, while neuroimaging directly visualizes the brain activation that it evokes in several cortical areas. This activation is often attenuated under subliminal presentation conditions compared to consciously reportable conditions, but there are sufficiently many exceptions, in paradigms such as the attentional blink, to indicate that high activation, per se, is not a sufficient condition for conscious access to occur. We conclude by arguing that for a stimulus to reach consciousness, two factors are jointly needed: (i) the input stimulus must have enough strength (which can be prevented by masking) and (ii) it must receive top-down attention (which can be prevented by drawing attention to another stimulus or task). This view leads to a distinction between two types of non conscious processes, which we call subliminal and preconscious. According to us, maintaining this distinction is essential in order to make sense of the growing neuroimaging data on the neural correlates of consciousness. PMID- 17403643 TI - The human amygdala and orbital prefrontal cortex in behavioural regulation. AB - Survival in complex environments depends on an ability to optimize future behaviour based on past experience. Learning from experience enables an organism to generate predictive expectancies regarding probable future states of the world, enabling deployment of flexible behavioural strategies. However, behavioural flexibility cannot rely on predictive expectancies alone and options for action need to be deployed in a manner that is responsive to a changing environment. Important moderators on learning-based predictions include those provided by context and inputs regarding an organism's current state, including its physiological state. In this paper, I consider human experimental approaches using functional magnetic resonance imaging that have addressed the role of the amygdala and prefrontal cortex (PFC), in particular the orbital PFC, in acquiring predictive information regarding the probable value of future events, updating this information, and shaping behaviour and decision processes on the basis of these value representations. PMID- 17403644 TI - Function and localization within rostral prefrontal cortex (area 10). AB - We propose that rostral prefrontal cortex (PFC; approximating area 10) supports a cognitive system that facilitates either stimulus-oriented (SO) or stimulus independent (SI) attending. SO attending is the behaviour required to concentrate on current sensory input, whereas SI attending is the mental processing that accompanies self-generated or self-maintained thought. Regions of medial area 10 support processes related to the former, whilst areas of lateral area 10 support processes that enable the latter. Three lines of evidence for this 'gateway hypothesis' are presented. First, we demonstrate the predicted patterns of activation in area 10 during the performance of new tests designed to stress the hypothetical function. Second, we demonstrate area 10 activations during the performance of established functions (prospective memory, context memory), which should hypothetically involve the proposed attentional system. Third, we examine predictions about behaviour-activation patterns within rostral PFC that follow from the hypothesis. We show with meta-analysis of neuroimaging investigations that these predictions are supported across a wide variety of tasks, thus establishing a general principle for functional imaging studies of this large brain region. We then show that while the gateway hypothesis accommodates a large range of findings relating to the functional organization of area 10 along a medial-lateral dimension, there are further principles relating to other dimensions and functions. In particular, there is a functional dissociation between the anterior medial area 10, which supports processes required for SO attending, and the caudal medial area 10, which supports processes relating to mentalizing. PMID- 17403645 TI - Drying and wetting of Mediterranean soils stimulates decomposition and carbon dioxide emission: the "Birch effect". AB - Observations on the net carbon exchange of forests in the European Mediterranean region, measured recently by the eddy covariance method, have revived interest in a phenomenon first characterized on agricultural and forest soils in East Africa in the 1950s and 1960s by H. F. Birch and now often referred to as the "Birch effect." When soils become dry during summer because of lack of rain, as is common in regions with Mediterranean climate, or are dried in the laboratory in controlled conditions, and are then rewetted by precipitation or irrigation, there is a burst of decomposition, mineralization and release of inorganic nitrogen and CO(2). In forests in Mediterranean climates in southern Europe, this effect has been observed with eddy covariance techniques and soil respiration chambers at the stand and small plot scales, respectively. Following the early work of Birch, laboratory incubations of soils at controlled temperatures and water contents have been used to characterize CO(2) release following the rewetting of dry soils. A simple empirical model based on laboratory incubations demonstrates that the amount of carbon mineralized over one year can be predicted from soil temperature and precipitation regime, provided that carbon lost as CO(2) is taken into account. We show that the amount of carbon returned to the atmosphere following soil rewetting can reduce significantly the annual net carbon gain by Mediterranean forests. PMID- 17403646 TI - Ozone air pollution effects on tree-ring growth, delta(13)C, visible foliar injury and leaf gas exchange in three ozone-sensitive woody plant species. AB - We assessed the effects of ambient tropospheric ozone on annual tree-ring growth, delta(13)C in the rings, leaf gas exchange and visible injury in three ozone sensitive woody plant species in southern Switzerland. Seedlings of Populus nigra L., Viburnum lantana L. and Fraxinus excelsior L. were exposed to charcoal filtered air (CF) and non-filtered air (NF) in open-top chambers, and to ambient air (AA) in open plots during the 2001 and 2002 growing seasons. Ambient ozone exposures in the region were sufficient to cause visible foliar injury, early leaf senescence and premature leaf loss in all species. Ozone had significant negative effects on net photosynthesis and stomatal conductance in all species in 2002 and in V. lantana and F. excelsior in 2001. Water-use efficiency decreased and intercellular CO(2) concentrations increased in all species in response to ozone in 2002 only. The width and delta(13)C of the 2001 and 2002 growth rings were measured for all species at the end of the 2002 growing season. Compared with CF seedlings, mean ring width in the AA and NF P. nigra seedlings was reduced by 52 and 46%, respectively, in 2002, whereas in V. lantana and F. excelsior, ring width showed no significant reductions in either year. Although delta(13)C was usually more negative in CF seedlings than in AA and NF seedlings, with the exception of F. excelsior in 2001, ozone effects on delta(13)C were significant only for V. lantana and P. nigra in 2001. Among species, P. nigra exhibited the greatest response to ozone for the measured parameters as well as the most severe foliar injury and was the only species to show a significant reduction in ring width in response to ozone exposure, despite significant negative ozone effects on leaf gas exchange and the development of visible foliar injury in V. lantana and F. excelsior. Thus, significant ozone-induced effects at the leaf level did not correspond to reduced tree-ring growth or increased delta(13)C in all species, indicating that the timing of ozone exposure and severity of leaf-level responses may be important in determining the sensitivity of tree productivity to ozone exposure. PMID- 17403647 TI - Leaf exchange in a Mediterranean shrub: water, nutrient, non-structural carbohydrate and osmolyte dynamics. AB - Leaf exchange is an abrupt phenological event that drastically modifies the morphology and physiology of the aerial portion of the plant. We examined if water and osmolyte differences between old leaves and new organs trigger leaf exchange, and whether the differences are closely linked to the resource resorption process in senescing leaves. We monitored concentrations of osmolyte, water, non-structural carbohydrate, nitrogen and potassium in senescing leaves and in emerging new leaves and inflorescences of a Mediterranean leaf exchanger (Cistus laurifolius L.) growing in NE Spain. Old leaves rehydrated markedly during most of the senescence process, which co-occurred with the extension of new shoots, suggesting the lack of a clear-cut switch in water supply from old to new organs. The accumulation of osmolytes in the early stage of leaf senescence might account for this rehydration. Osmolyte dynamics in old leaves depended largely on the progression of resource resorption from senescing organs but were mostly unrelated to water content during late senescence. We conclude that dehydration of old leaves is not a prerequisite for the triggering of leaf exchange. The finding that most nutrients and carbohydrates accumulated in new organs before senescing leaves massively exported resources, and the absence of relevant differences between the dynamics of old leaves at the base of inflorescences and those at the base of vegetative shoots, indicate that the nutrient and carbohydrate demands of new organs do not trigger leaf exchange. PMID- 17403648 TI - Pacific Ocean and Japan Sea ecotypes of Japanese beech (Fagus crenata) differ in photosystem responses to continuous high light. AB - Two ecotypes of Japanese beech (Fagus crenata Blume), the Pacific Ocean type (PAO) and the Japan Sea type (JAS), show different responses to high solar irradiance. When PAO and JAS saplings were grown in continuous high-light (H), leaves of JAS became pale green. To elucidate this phenomenon, we investigated in vivo photochemistry based on pigment concentrations of Photosystem (PS) I and PS II and Western blot analysis. In JAS-H leaves, the amount of D1-protein decreased, resulting in decreases in the maximal quantum yield of PS II (F(v)/F(m)) and electron transport rate, whereas PAO-H leaves maintained high activities. The PS I photochemistry determined by measurement of P-700 photo oxidation showed that the intersystem electron pool size was 1.4 times greater in JAS-H leaves than in PAO-H leaves. Furthermore, the re-reduction kinetics of P 700(+) showed that cyclic electron transport around PS I was 1.2 times faster in PAO-H leaves than in JAS-H leaves. Analysis of the area over the fluorescence induction kinetics indicated that the relative abundance of the PS IIalpha center increased in PAO-H leaves, whereas JAS leaves were observed to have low acclimation capacity to high light. These results demonstrate that PAO leaves possess acclimation mechanisms to continuous high light, whereas JAS leaves are more vulnerable to continuous high light, resulting in reduced leaf longevity owing to photoinhibition caused by increases in the intersystem electron pool size and suppression of photochemistry at the level of PS I and PS II. PMID- 17403649 TI - Correlation between acoustic emission, water status and xylem embolism in pine wilt disease. AB - The occurrence of cavitation events and embolism during the latent, early stage and the late developmental stages of pine wilt disease was monitored nondestructively by acoustic emission (AE) and high-resolution magnetic resonance microscopy, respectively, and the results were compared with changes in leaf water potential and stem thickness. In the latent stage of the disease, when no embolisms were observed, cavitation events were detected by AE during the daytime in water-stressed Japanese black pine (Pinus thunbergii Parl.) seedlings, indicating that cavitation occurred at the individual tracheid level. In the early stage of the disease, an increase in the frequency of AE events occurred coincidentally with the occurrence of patchy embolisms at the mass tracheid level. The threshold water potential for such mass cavitation was higher than that causing cavitation of individual tracheids during the latent stage of the disease. In the advanced stage of the disease, explosive AE events were observed coincidentally with drastic enlargement of embolized areas and decreases in water potential. The AE events in the latent stage occurred only during the daytime, whereas, in the early and advanced stages of the disease, they also occurred at night. The explosive occurrence of cavitation in the advanced stage was thought to be a case of "runaway embolism." PMID- 17403650 TI - Photosynthetic characteristics of olive tree (Olea europaea) bark. AB - Functional and structural characteristics of corticular photosynthesis of sun exposed bark of olive tree (Olea europaea L.) were examined. Stomata are only sporadically present during stem primary growth. Light transmission through the phellem was age dependent, decreasing rapidly in stems older than five years of age. Light transmission was also low in pubescent 1-year-old stems. Light transmission was about 50% higher in wet phellem than in dry phellem. Photosynthetic capacity on a unit area basis (measured with an oxygen disc electrode at 27 degrees C and about 5% CO(2) on chlorophyllous tissue discs isolated from the stem) was higher in 1-, 20- and 30-year-old stems compared with 2-10-year-old stems. Low chlorophyll a/b ratio and light compensation points were recorded in olive stems with low phellem light transmission, in accordance with the shade acclimation hypothesis. The intrinsic photochemical efficiency of photosystem II of all stems, especially young stems, was less than that of the leaves. Our results show that olive tree bark possesses an efficient photosynthetic mechanism that may significantly contribute not only to the reduction in concentrations of CO(2) in the inner bark, but also to whole-tree carbon balance. PMID- 17403651 TI - Seasonal change in the drought response of wood cell development in poplar. AB - Field-grown poplar trees (Populus nigra L. x P. maximowiczii Henry, clone Kamabuchi) were exposed to severe drought twice during the growing season to evaluate the impact on wood cell development. The drought treatment caused a reduction in leaf water potential, leaf wilting and a decreased concentration of osmotically active solutes in the cambial zone. Drought-induced changes in the anatomy of developing xylem cells were examined in stem sections and macerated wood samples. In early summer, drought significantly reduced the length and cross sectional area of newly formed fibers, whereas no such effects were observed in late summer. In well-watered trees, fiber cross-sectional area declined between early and late summer. Similarly, drought reduced the cross-sectional area of vessel elements in early summer but not in late summer, whereas in both control and drought-treated trees, the cross-sectional area of vessel elements decreased between early and late summer. The vessel area to xylem area ratio was unaffected by drought because the drought-induced decrease in vessel size was matched by an increase in the number of newly formed vessel cells. In contrast to its effect in early summer, late-summer drought had no significant effect on fiber and vessel cell development, indicating that sensitivity of wood cell development to drought varies seasonally. PMID- 17403652 TI - Optimal conditions for visualizing water-conducting pathways in a living tree by the dye injection method. AB - To elucidate the water-conducting pathways in living trees by the dye injection method, suitable sample preparation procedures are needed. We evaluated quantitatively the properties and concentrations of three dyes (acid fuchsin, basic fuchsin and safranin) widely used for this purpose, and determined the optimal conditions required to avoid artifacts after dye injection into the sap stream of Pieris japonica D. Don. Among the dyes tested, an aqueous solution of acid fuchsin at a concentration of 0.1% or more was the most useful for delineating water movement. In non-transpiring stem segments, the vertical movement of acid fuchsin by capillarity and diffusion from the dye injection site was limited. However, acid fuchsin moved rapidly in the horizontal direction by capillarity and diffusion, and most xylem cells were stained within 2 h. A delay of more than 2 h between dye injection and examination of the tissues greatly reduces the precision of the method. Use of the dye injection method without appropriate, well-defined experimental procedures may give rise to misleading information about the functional water-conducting pathway in living trees. PMID- 17403653 TI - Can fog contribute to the nutrition of Chamaecyparis obtusa var. formosana? Uptake of a fog solute tracer into foliage and transport to roots. AB - Yellow cypress (Chamaecyparis obtusa (Siebold & Zucc.) Endl. var. formosana (Hayata) Rehder) is the predominant tree species of Taiwan's nutrient-poor, mountain fog forests. Little is known about the potential contribution of solute uptake from fog to the overall nutrition of these trees. Shoots of yellow cypress seedlings were misted with artificial fog containing the tracer rubidium (Rb) in laboratory and field experiments to determine if there is solute uptake from the fog. After misting shoots for six weeks, substantial amounts of tracer were detected in unexposed roots by inductively coupled plasma mass spectroscopy bulk analysis. Possible routes of entry were examined by element imaging with energy dispersive X-ray analysis. Direct uptake of the tracer into leaves across the cuticle and epidermis was small, excluding this as the major uptake path. Accumulations of Rb were found on leaf surfaces along the edges of the leaves. The almost daily changes in fog coverage and air humidity may enhance the accumulation of fog solutes at leaf edges. Accumulation of Rb was also found in narrow clefts between opposite leaves and between the outermost and underlying alternating stacked leaves. The clefts provide a direct passage from the leaf surface to the space beneath the imbricate leaves and the underlying alternate leaves, possibly facilitating solute uptake from fog, which in turn may contribute to the nutrition of yellow cypress. PMID- 17403654 TI - Water relations of seedlings of three Quercus species: variations across and within species grown in contrasting light and water regimes. AB - We compared seedling water relations of three Mediterranean Quercus species (the evergreen shrub Q. coccifera L., the evergreen tree Q. ilex L. subsp. ballota (Desf.) Samp. and the deciduous or marcescent tree Q. faginea L.). We also explored seedling potential for acclimation to contrasting growing conditions. In March, 1-year-old seedlings of the three species were planted in pots and grown outdoors in a factorial combination of two irrigation regimes (daily (HW) and alternate day watering (LW)) and two irradiances (43 and 100% of full sunlight). At the end of July, predawn and midday water potentials (Psi(pd), Psi(md)) were measured, and pressure-volume (P-V) curves were obtained for mature current-year shoots. Species exhibited similar Psi(pd) and Psi(md) values, but differed in leaf morphology and water relations. The evergreens possessed larger leaf mass per area (LMA) and were able to maintain positive turgor pressure at lower water potentials than the deciduous species because of their lower osmotic potential at full turgor. However, the three species had similar relative water contents at the turgor loss point because Q. faginea compensated for its higher osmotic potential with greater cell wall elasticity. Values of Psi(pd) had a mean of 1.12 MPa in LW and -0.63 MPa in HW, and Psi(md) had a mean of -1.13 MPa in full sunlight and -1.64 MPa in shade, where seedlings exhibited lower LMA. However, the P-V curve traits were unaffected by the treatments. Our results suggest that Q. faginea seedlings combine the water-use characteristics of mesic deciduous oak and the drought-tolerance of xeric evergreen oak. The ability of Q. coccifera to colonize drier sites than Q. ilex was not a result of higher drought tolerance, but rather may be associated with other dehydration postponement mechanisms including drought-induced leaf shedding. The lack of treatment effects may reflect a relatively low contrast between treatment regimes, or a low inherent responsiveness of these traits in the study species, or both. PMID- 17403655 TI - Temperature sum accumulation effects on within-population variation and long-term trends in date of bud burst of European white birch (Betula pendula). AB - Within-population variation in phenology of boreal trees indicates their adaptability to climatic variations. Although interannual variations in date of bud burst have been widely discussed, little is known about within-population variation, the key determinants for this variation and the effects of this variation on estimates of trends in bud burst date. Over a period of nine years, we monitored timing of bud burst daily in 30 mature white birch (Betula pendula Roth) trees in a naturally regenerated stand. Our results revealed not only large interannual variation but also considerable intraannual variation among individual trees in date of bud burst, the maximum within-population variation being four weeks. Bud burst can be accurately predicted by the date when a threshold value of temperature sum in spring is reached (base temperature +5 degrees C). Based on this temperature sum and past temperature records, we estimated the trend in date of bud burst. The linear trend estimate based on the years 1926-2005 is an advancement of 1.2 days per decade (95% confidence interval, +/- 0.7 days), which is much less than that predicted by time series based on coarser time intervals. We conclude that, because of large interannual differences, and large annual within-population variations in bud burst, estimates of bud burst date based on measurements made over a period of only a few decades are unreliable. PMID- 17403656 TI - Intra- and interspecific diversity in the response to waterlogging of two co occurring white oak species (Quercus robur and Q. petraea). AB - Quercus robur L. and Quercus petraea (Matt.) Liebl. are sympatric oak species with different ecological requirements. Quercus robur is more tolerant to waterlogging than Q. petraea. This ecological divergence may play a role in the maintenance of the two species despite the absence of an insurmountable reproductive barrier between them. We predicted that the genetic architecture of traits related to waterlogging tolerance differs between the species. To gain insight into this architecture in the absence of genetic markers for waterlogging tolerance, we compared populations of seedlings of each species for diversity in the expression of quantitative phenotypic traits induced by severe hypoxia. To determine the capacity for hypertrophied lenticel formation, we applied gas impermeable mastic to stems of seedlings. Two months after application, the mastic treatment had induced the formation of 3 (+/- 2) cm(-2) hypertrophied lenticels in the absence of root hypoxia. Leaf epinasty during root hypoxia was an early predictor of seedling mortality. Four weeks of waterlogging resulted in greater epinasty in Q. petraea than in Q. robur, but fewer hypertrophied lenticels (16 +/- 6 versus 21 +/- 9 cm(-2)) and adventitious roots (2.7 +/- 4.7 versus 5.2 +/- 5.9). Differences between species in these traits were associated with differences in the frequencies of extreme phenotypes rather than with a generally higher tolerance to waterlogging in Q. robur seedlings. PMID- 17403657 TI - Seasonal changes in the temperature response of photosynthesis in canopy leaves of Quercus crispula in a cool-temperate forest. AB - Understanding seasonal changes in photosynthetic characteristics of canopy leaves is indispensable for modeling the carbon balance in forests. We studied seasonal changes in gas exchange characteristics that are related to the temperature dependence of photosynthesis in canopy leaves of Quercus crispula Blume, one of the most abundant species in cool-temperate forests in Japan. Photosynthetic rate and ribulose-1,5-bisphosphate (RuBP) carboxylation capacity (V(cmax)) at 20 degrees C increased from June to August and then decreased in September. The activation energy of V(cmax), a measure of the temperature dependence of V(cmax), was highest in summer, indicating that V(cmax) was most sensitive to leaf temperature at this time. The activation energy of V(cmax) was significantly correlated with growth temperature. Other parameters related to the temperature dependence of photosynthesis, such as intercellular CO(2) partial pressure and temperature dependence of RuBP regeneration capacity, showed no clear seasonal trend. It was suggested that leaf senescence affected the balance between carboxylation and regeneration of RuBP. The model simulation showed that photosynthetic rate and its optimal temperature were highest in summer. PMID- 17403658 TI - Roles of soil chemistry and water availability in site-related delta(13)C variations in French beech forests. AB - The carbon isotopic composition (delta(13)C) of wood and leaf cellulose of beech trees (Fagus sylvatica L.) was studied at 80 sites in northeastern France. We sampled sites with contrasting water balance, depending on soil type and precipitation. We tested the hypothesis that inter-site variations in plant delta(13)C reflect the spatial distribution of soil water availability, and we assessed whether delta(13)C could be used as a bioindicator of soil water availability. Patterns of variation in delta(13)C were compared with estimates of monthly water balance and with other soil characteristics. Between-site variability in delta(13)C was high (2.9 per thousand range in wood cellulose, 2.1 per thousand in leaf cellulose), but variation in water availability appeared to be only a minor factor contributing to this variation in delta(13)C. Unexpectedly, spatial variations in wood and leaf cellulose delta(13)C were significantly and positively related to soil fertility expressed by soil pH (r = 0.42 and 0.43, respectively) and cation content. On average, trees growing on acidic soils displayed 0.5 per thousand lower delta(13)C in both wood and leaf material than trees growing on neutral or calcareous soils. Our initial hypothesis of a strong negative relationship between delta(13)C and site water availability was not confirmed. In the study zone, neither wood nor leaf delta(13)C appeared to be a reliable bioindicator of spatial variations in water availability. Possible causes for the lack of a relationship are discussed. Our findings confirm, under natural conditions, the strong effect of soil fertility on water-use efficiency previously observed in experiments. This effect needs to be considered in isotopic studies involving different sites. PMID- 17403659 TI - Defoliation and nitrogen effects on photosynthesis and growth of Eucalyptus globulus. AB - Plant responses to defoliation are complex. We established a field experiment in a nine-month-old Eucalyptus globulus Labill. plantation to examine the effects of pattern (upper crown versus lower crown removal), frequency (single, double or triple defoliation within a 12-month period) and severity (25 versus 38% of leaf area removed) of defoliation and the effect of soil nitrogen (N) on photosynthetic processes and stem growth. The photosynthetic responses observed following defoliation could be attributed to changes in source:sink ratios. Light saturated CO(2) uptake (A(max)) increased with increasing severity and frequency of defoliation irrespective of defoliation pattern. Seedlings defoliated in autumn did not exhibit increases in A(max) until the following spring, whereas there was no such delay in photosynthetic responses associated with spring defoliation. Application of N before defoliation allowed trees to compensate for the effect of defoliation on stem diameter growth, which could not be explained simply in terms of increases in A(max). The observed increases in stem diameter increment following N fertilization of defoliated trees suggested increases in leaf area development, and there were changes in the leaf area:leaf dry mass ratio that may have increased light absorption by the crown. Nitrogen fertilization also increased partitioning of dry mass to branches at the expense of main stems, suggesting that N supply was important in rebuilding crowns following a defoliation event. PMID- 17403660 TI - Carbon isotope discrimination and water stress in trembling aspen following variable retention harvesting. AB - Variable retention harvesting (VRH) has been proposed as a silvicultural practice to maintain biodiversity and ecosystem integrity. No previous study has examined tree carbon isotope discrimination to provide insights into water stress that could lead to dieback and mortality of trees following VRH. We measured and compared the carbon isotope ratios (delta(13)C) in stem wood of trembling aspen (Populus tremuloides Michx.) before and after VRH. Eight trees were sampled from isolated residual, edge and control (interior of unharvested stand) positions from each of seven plots in three regions (Calling Lake and Drayton Valley, Alberta and Lac Duparquet, Quebec). After VRH, the general trend in mean delta(13)C was residual > edge > control trees. Although this trend is indicative of water stress in residual trees, it also suggests that edge trees received some sheltering effect, reducing their stress compared with that of residuals. A strong inverse relationship was found between the delta(13)C values and the mean annual precipitation in each region. The trend in mean delta(13)C signature was Calling Lake > Drayton Valley > Lac Duparquet trees. These results suggest that residual or edge trees in drier regions are more likely to suffer water stress following VRH. We also observed a trend of greater delta(13)C in stout trees compared with slender trees, both before and after VRH. The evidence of greater water stress in stout trees likely occurred because of a positive relationship between stem diameter and crown volume per basal area. Our results provide evidence that water stress could be the driving mechanism leading to dieback and mortality of residual trees shortly after VRH. Additionally, the results from edge trees indicate that leaving hardwood residuals in larger patches or more sheltered landscape positions could reduce the water stress to which these trees are subjected, thereby reducing dieback and mortality. PMID- 17403662 TI - The contributions of integrin affinity and integrin-cytoskeletal engagement in endothelial and smooth muscle cell adhesion to vitronectin. AB - The serine proteinase inhibitor, plasminogen activator inhibitor type-1 (PAI-1), binds to the adhesion protein vitronectin with high affinity at a site that is located directly adjacent to the vitronectin RGD integrin binding sequence. The binding of PAI-1 to vitronectin sterically blocks integrin access to this site and completely inhibits the binding of purified integrins to vitronectin; however, its inhibition of endothelial and smooth muscle cell adhesion to vitronectin is at most 50-75%. Because PAI-1 binds vitronectin with approximately 10-100-fold higher affinity than purified integrins, we have analyzed the mechanism whereby these cells are able to overcome this obstacle. Our studies exclude proteolytic removal of PAI-1 from vitronectin as the mechanism, and show instead that cell adhesion in the presence of PAI-1 is dependent on integrin cytoskeleton engagement. Disrupting endothelial or smooth muscle cell actin polymerization and/or focal adhesion assembly reduces cell adhesion to vitronectin in the presence of PAI-1 to levels similar to that observed for the binding of purified integrins to vitronectin. Furthermore, endothelial cell, but not smooth muscle cell adhesion to vitronectin in the presence of PAI-1 requires both polymerized microtubules and actin, further demonstrating the importance of the cytoskeleton for integrin-mediated adhesion. Finally, we show that cell adhesion in the presence of PAI-1 leads to colocalization of PAI-1 with the integrins alphavbeta3 and alphavbeta5 at the cell-matrix interface. PMID- 17403661 TI - NFATc3 mediates chronic hypoxia-induced pulmonary arterial remodeling with alpha actin up-regulation. AB - Physiological responses to chronic hypoxia include polycythemia, pulmonary arterial remodeling, and vasoconstriction. Chronic hypoxia causes pulmonary arterial hypertension leading to right ventricular hypertrophy and heart failure. During pulmonary hypertension, pulmonary arteries exhibit increased expression of smooth muscle-alpha-actin and -myosin heavy chain. NFATc3 (nuclear factor of activated T cells isoform c3), which is aCa(2+)-dependent transcription factor, has been recently linked to smooth muscle phenotypic maintenance through the regulation of the expression of alpha-actin. The aim of this study was to determine if: (a) NFATc3 is expressed in murine pulmonary arteries, (b) hypoxia induces NFAT activation, (c) NFATc3 mediates the up-regulation of alpha-actin during chronic hypoxia, and (d) NFATc3 is involved in chronic hypoxia-induced pulmonary vascular remodeling. NFATc3 transcript and protein were found in pulmonary arteries. NFAT-luciferase reporter mice were exposed to normoxia (630 torr) or hypoxia (380 torr) for 2, 7, or 21 days. Exposure to hypoxia elicited a significant increase in luciferase activity and pulmonary arterial smooth muscle nuclear NFATc3 localization, demonstrating NFAT activation. Hypoxia induced up regulation of alpha-actin and was prevented by the calcineurin/NFAT inhibitor, cyclosporin A (25 mg/kg/day s.c.). In addition, NFATc3 knock-out mice did not showed increased alpha-actin levels and arterial wall thickness after hypoxia. These results strongly suggest that NFATc3 plays a role in the chronic hypoxia induced vascular changes that underlie pulmonary hypertension. PMID- 17403663 TI - A quantitative kinetic model for the in vitro assembly of intermediate filaments from tetrameric vimentin. AB - In vitro assembly of intermediate filament proteins is a very rapid process. It starts without significant delay by lateral association of tetramer complexes into unit-length filaments (ULFs) after raising the ionic strength from low salt to physiological conditions (100 mM KCl). We employed electron and scanning force microscopy complemented by mathematical modeling to investigate the kinetics of in vitro assembly of human recombinant vimentin. From the average length distributions of the resulting filaments measured at increasing assembly times we simulated filament assembly and estimated specific reaction rate parameters. We modeled eight different potential pathways for vimentin filament elongation. Comparing the numerical with the experimental data we conclude that a two-step mechanism involving rapid formation of ULFs followed by ULF and filament annealing is the most robust scenario for vimentin assembly. These findings agree with the first two steps of the previously proposed three-step assembly model (Herrmann, H., and Aebi, U. (1998) Curr. Opin. Struct. Biol. 8, 177-185). In particular, our modeling clearly demonstrates that end-to-end annealing of ULFs and filaments is obligatory for forming long filaments, whereas tetramer addition to filament ends does not contribute significantly to filament elongation. PMID- 17403664 TI - Def-6, a guanine nucleotide exchange factor for Rac1, interacts with the skeletal muscle integrin chain alpha7A and influences myoblast differentiation. AB - Integrin alpha7beta1 is the major laminin binding integrin receptor of muscle cells. The alpha7 chain occurs in several splice isoforms, of which alpha7A and alpha7B differ in their intracellular domains only. The fact that the expression of alpha7A and alpha7B is tightly regulated during skeletal muscle development suggests different and distinct roles for both isoforms. However, so far, functional properties and interacting proteins were described for the alpha7B chain only. Using a yeast two-hybrid screen, we have found that Def-6, a guanine nucleotide exchange factor for Rac1, binds to the intracellular domain of the alpha7A subunit. The specificity of the Def-6-alpha7A interaction has been shown by direct yeast two-hybrid binding assays and coprecipitation experiments. This is the first description of an alpha7A-specific and -exclusive interaction, because Def-6 did not bind to any other tested integrin cytoplasmic domain. Interestingly, the binding of Def-6 to alpha7A was abolished, when cells were cotransfected with an Src-related kinase, which is known to phosphorylate Def-6 and stimulate its exchange activity. We found expression of Def-6 was not only restricted to T-lymphocytes as described thus far but in a more widespread manner, including different muscle tissues. In cells, Def-6 is seen in newly forming cell protrusions and focal adhesions, and its localization partially overlaps with the alpha7A integrin receptor. C2C12 myoblasts overexpressing Def-6 show a delay of Rac1 inactivation during myogenic differentiation and abnormal myotube formation. Thus, our data suggest a role for Def-6 in the fine regulation of Rac1 during myogenesis with the integrin alpha7A chain guiding this regulation in a spatio-temporal manner. PMID- 17403665 TI - Unique peroxidase reaction mechanism in prostaglandin endoperoxide H synthase-2: compound I in prostaglandin endoperoxide H synthase-2 can be formed without assistance by distal glutamine residue. AB - Prostaglandin-endoperoxide H synthase-2 (PGHS-2) shows peroxidase activity to promote the cyclooxygenase reaction for prostaglandin H2, but one of the highly conserved amino acid residues in peroxidases, distal Arg, stabilizing the developing negative charge on the peroxide through a hydrogen-bonding interaction, is replaced with a neutral amino acid residue, Gln. To characterize the peroxidase reaction in PGHS-2, we prepared three distal glutamine (Gln-189) mutants, Arg (Gln-->Arg), Asn (Gln-->Asn), and Val (Gln-->Val) mutants, and examined their peroxidase activity together with their structural characterization by absorption and resonance Raman spectra. Although a previous study (Landino, L. M., Crews, B. C., Gierse, J. K., Hauser, S. D., and Marnett, L. (1997) J. Biol. Chem. 272, 21565-21574) suggested that the Gln residue might serve as a functionally equivalent residue to Arg, our current results clearly showed that the peroxidase activity of the Val and Asn mutants was comparable with that of the wild-type enzyme. In addition, the Fe-C and C-O stretching modes in the CO adduct were almost unperturbed by the mutation, implying that Gln-189 might not directly interact with the heme-ligated peroxide. Rather, the peroxidase activity of the Arg mutant was depressed, concomitant with the heme environmental change from a six-coordinate to a five-coordinate structure. Introduction of the bulky amino acid residue, Arg, would interfere with the ligation of a water molecule to the heme iron, suggesting that the side chain volume, and not the amide group, at position 189 is essential for the peroxidase activity of PGHS-2. Thus, we can conclude that the O-O bond cleavage in PGHS-2 is promoted without interactions with charged side chains at the peroxide binding site, which is significantly different from that in typical plant peroxidases. PMID- 17403666 TI - Isolation and characterization of proteins associated with histone H3 tails in vivo. AB - The histone H3 amino-terminal tails play an important role in regulating chromatin transcription. Although the mechanisms by which the H3 tail modulates transcription are not well understood, recent discoveries of specific interactions of regulatory factors with H3 tails suggest that H3 tails are a key player in the precise regulation of transcription activity. To investigate the recruitment-based action of H3 tails in chromatin transcription, we purified H3 tail-associated proteins from HeLa cells that stably express epitope-tagged H3 tails. This approach resulted in the identification of multiple histone methyltransferase activities and transcription regulatory factors that are specifically associated with expressed H3 tail domains. Point mutations of Lys-9 and Lys-27 to block cellular modifications of the tail domains completely abolished the association of specific factors, including HP1 and several repressors. Importantly, our transcription analysis revealed that the purified factors can significantly stimulate p300-mediated transcription from chromatin templates. These results implicate that the H3 tail, when accessible in relaxed chromatin, acts as a transcriptional regulator by mediating recruitment of specific sets of cofactors. PMID- 17403667 TI - Systematic analysis of the entire second extracellular loop of the V(1a) vasopressin receptor: key residues, conserved throughout a G-protein-coupled receptor family, identified. AB - The roles of extracellular residues of G-protein-coupled receptors (GPCRs) are not well defined compared with residues in transmembrane helices. Nevertheless, it has been established that extracellular domains of both peptide-GPCRs and amine-GPCRs incorporate functionally important residues. Extracellular loop 2 (ECL2) has attracted particular interest, because the x-ray structure of bovine rhodopsin revealed that ECL2 projects into the binding crevice within the transmembrane bundle. Our study provides the first comprehensive investigation into the role of the individual residues comprising the entire ECL2 domain of a small peptide-GPCR. Using the V(1a) vasopressin receptor, systematic substitution of all of the ECL2 residues by Ala generated 30 mutant receptors that were characterized pharmacologically. The majority of these mutant receptor constructs (24 in total) had essentially wild-type ligand binding and intracellular signaling characteristics, indicating that these residues are not critical for normal receptor function. However, four aromatic residues Phe(189), Trp(206), Phe(209), and Tyr(218) are important for agonist binding and receptor activation and are highly conserved throughout the neurohypophysial hormone subfamily of peptide-GPCRs. Located in the middle of ECL2, juxtaposed to the highly conserved disulfide bond, Trp(206) and Phe(209) project into the binding crevice. Indeed, Phe(209) is part of the Cys-X-X-X-Ar (where Ar is an aromatic residue) motif, which is well conserved in both peptide-GPCRs and amine-GPCRs. In contrast, Phe(189) and Tyr(218), located at the extreme ends of ECL2, may be important for determining the position of the ECL2 cap over the binding crevice. This study provides mechanistic insight into the roles of highly conserved ECL2 residues. PMID- 17403668 TI - Kaposi sarcoma-associated herpes virus targets the lymphotactin receptor with both a broad spectrum antagonist vCCL2 and a highly selective and potent agonist vCCL3. AB - Large DNA viruses such as herpesvirus and poxvirus encode proteins that target and exploit the chemokine system of their host. These proteins have the potential to block or change the orchestrated recruitment of leukocytes to sites of viral infection. The genome of Kaposi sarcoma-associated herpes virus (KSHV) encodes three chemokine-like proteins named vCCL1, vCCL2, and vCCL3. In this study vCCL3 was probed in parallel with vCCL1 and vCCL2 against a panel of the 18 classified human chemokine receptors. In calcium mobilization assays vCCL1 acted as a selective CCR8 agonist, whereas vCCL2 was found to act as a broad spectrum chemokine antagonist of human chemokine receptors, including the lymphotactin receptor. In contrast vCCL3 was found to be a highly selective agonist for the human lymphotactin receptor XCR1. The potency of vCCL3 was found to be 10-fold higher than the endogenous human XCL1 chemokine in respect to phosphatidylinositol turnover and calcium mobilization as well as chemotaxis. High expression of XCR1 was found in placenta and neutrophils by real-time PCR. These data are consistent with reports of different expression profiles for vCCL2 and vCCL3 during the life cycle of KSHV, indicate a novel, sophisticated exploitation by the virus of specifically the lymphotactin receptor by both agonist and antagonist mechanisms, and suggest a unique physiological importance of this (somewhat overlooked) chemokine receptor. PMID- 17403669 TI - Glucose mediates the translocation of NeuroD1 by O-linked glycosylation. AB - O-Linked GlcNAc modification of nuclear and cytosolic proteins has been shown to regulate the function of many cellular proteins. Increased O-linked glycosylation, observed under chronic hyperglycemia conditions, has been implicated in the pathogenesis of diabetes. However, the exact role of O-GlcNAc modification in regulating glucose homeostasis remains to be established. We report here that the subcellular localization of the pancreatic beta cell specific transcription factor NeuroD1 is regulated by O-linked glycosylation in the mouse insulinoma cell line MIN6. Under low glucose conditions, NeuroD1 is mainly in the cytosol. However, treatment of MIN6 cells with high glucose results in O-linked GlcNAc modification of NeuroD1 and its subsequent translocation into the nucleus. Consistent with these data, treatment of MIN6 cells with O-(2 acetamido-2-deoxy-d-glucopyranosylidene)-amino N-phenylcarbamate, an inhibitor of O-GlcNAcase, causes Neuro-D1 localization to the nucleus and induction of insulin gene expression even on low glucose. Furthermore, we demonstrate that NeuroD1 interacts with the O-GlcNAc transferase, OGT only at high concentrations of glucose and depletion of OGT by using small interfering RNA oligos interferes with the nuclear localization of NeuroD1 on high glucose. On low glucose NeuroD1 interacts with the O-GlcNAcase and becomes deglycosylated, which is likely to be important for export of Neuro-D1 into cytosol in the presence of low glucose. In summary, the presented data suggest that glucose regulates the subcellular localization of NeuroD1 in pancreatic beta cells via O-linked GlcNAc modification of NeuroD1 by OGT. PMID- 17403670 TI - Cell cycle-dependent expression of centrosomal ninein-like protein in human cells is regulated by the anaphase-promoting complex. AB - The recently identified centrosome protein Nlp (ninein-like protein) is a key regulator in centrosome maturation, which contributes to chromosome segregation and cytokinesis. However, the mechanism(s) controlling Nlp expression remains largely unknown. Here we have shown that Nlp expression is cell cycle-dependent with a peak at G(2)/M transition in human cells. Nlp is a short-lived protein and degraded by the proteasome via the anaphase-promoting cyclosome complex (APC/c) pathway. It interacts with the APC/c through the APC2 or Cdc27 subunits and is ubiquitinated. Following treatment with proteasome inhibitors, its protein level is elevated. Nlp binds in vivo to the degradation-targeting proteins Cdh1 and Cdc20, and overexpression of Cdh1 and Cdc20 enhances Nlp degradation. Using point mutations of the two putative degradation signals in Nlp, we have found that its degradation requires intact KEN-box and D-box. Interestingly, the Lys-Glu-Asn-D box-mutated Nlp exhibits a much stronger capability of inducing anchorage independent growth and multinuclearity compared with the wild type Nlp. Taken together, these findings indicate that Nlp expression is cell cycle-dependent and regulated by APC-mediated protein degradation. PMID- 17403671 TI - Thiazole synthase from Escherichia coli: an investigation of the substrates and purified proteins required for activity in vitro. AB - Thiamine is biosynthesized by combining two heterocyclic precursors. In Escherichia coli and other anaerobes, one of the heterocycles, 4-methyl-5-(beta hydroxyethyl) thiazole phosphate, is biosynthesized from 1-deoxyxylulose-5 phosphate, tyrosine, and cysteine. Genetic evidence has identified thiH, thiG, thiS, and thiF as essential for thiazole biosynthesis in E. coli. In this paper, we describe the measurement of the thiazole phosphate-forming reaction using purified protein components. The activity is shown to require four proteins isolated as heterodimers: ThiGH and ThiFS. Reconstitution of the [4Fe-4S] cluster in ThiH was essential for activity, as was the use of ThiS in the thiocarboxylate form. Spectroscopic studies with ThiGH strongly suggested that S adenosylmethionine (AdoMet) bound to the [4Fe-4S] cluster, which became more susceptible to reduction to the +1 state. Assays of thiazole phosphate formation showed that, in addition to the proteins, Dxp, tyrosine, AdoMet, and a reductant were required. The analysis showed that no more than 1 mol eq of thiazole phosphate was formed per ThiGH. Furthermore, for each mole of thiazole-P formed, 1 eq of AdoMet and 1 eq of tyrosine were utilized, and 1 eq of 5'-deoxyadenosine was produced. These results demonstrate that ThiH is a member of the "radical AdoMet" family and support a mechanistic hypothesis in which AdoMet is reductively cleaved to yield a highly reactive 5'-deoxyadenosyl radical. This radical is proposed to abstract the phenolic hydrogen atom from tyrosine, and the resultant substrate radical cleaves to yield dehydroglycine, which is required by ThiG for the thiazole cyclization reaction. PMID- 17403672 TI - Constitutive/hypoxic degradation of HIF-alpha proteins by the proteasome is independent of von Hippel Lindau protein ubiquitylation and the transactivation activity of the protein. AB - The transcriptional activator complex HIF-1 plays a key role in the long term adaptation of cells and tissues to their hypoxic microenvironment by stimulating the expression of genes involved in angiogenesis and glycolysis. The expression of the HIF-1 complex is regulated by the levels of its HIF-alpha subunits that are degraded under normoxic conditions by the ubiquitin-proteasome system. Whereas this pathway of HIF-alpha protein degradation has been well characterized, little is known of their turnover during prolonged hypoxic conditions. Herein, we describe a pathway by which HIF-1alpha and HIF-2alpha proteins are constitutively degraded during hypoxia by the proteasome system, although without requirement of prior ubiquitylation. The constitutive/hypoxic degradation of HIF-alpha proteins is independent of the presence of VHL, binding to DNA, or the formation of a transcriptionally active HIF-1 complex. These results are further strengthened by the demonstration that HIF-alpha proteins are directly degraded in a reconstituted in vitro assay by the proteasome. Finally, we demonstrate that the persistent down-regulation of HIF-1alpha during prolonged hypoxia is mainly caused by a decreased production of the protein without change in its degradation rate. This constitutive, ubiquitin-independent proteasomal degradation pathway of HIF-alpha proteins has to be taken into account in understanding the biology as well as in the development of therapeutic interventions of highly hypoxic tumors. PMID- 17403673 TI - The metallo-beta-lactamase GOB is a mono-Zn(II) enzyme with a novel active site. AB - Metallo-beta-lactamases (MbetaLs) are zinc-dependent enzymes able to hydrolyze and inactivate most beta-lactam antibiotics. The large diversity of active site structures and metal content among MbetaLs from different sources has limited the design of a pan-MbetaL inhibitor. Here we report the biochemical and biophysical characterization of a novel MbetaL, GOB-18, from a clinical isolate of a Gram negative opportunistic pathogen, Elizabethkingia meningoseptica. Different spectroscopic techniques, three-dimensional modeling, and mutagenesis experiments, reveal that the Zn(II) ion is bound to Asp120, His121, His263, and a solvent molecule, i.e. in the canonical Zn2 site of dinuclear MbetaLs. Contrasting all other related MbetaLs, GOB-18 is fully active against a broad range of beta-lactam substrates using a single Zn(II) ion in this site. These data further enlarge the structural diversity of MbetaLs. PMID- 17403674 TI - NADPH oxidase NOX5-S mediates acid-induced cyclooxygenase-2 expression via activation of NF-kappaB in Barrett's esophageal adenocarcinoma cells. AB - We have shown that the NADPH oxidase NOX5-S may play an important role in the progression from Barrett's esophagus to esophageal adenocarcinoma (EA) by increasing cell proliferation and decreasing apoptosis. However, the mechanism of the acid-induced NOX5-S-mediated increase in cell proliferation is not known. We found that, in SEG1 EA cells, the acid-induced increase in prostaglandin E2 (PGE2) production was mediated by activation of cyclooxygenase-2 (COX2) but not by COX1. Acid treatment increased intracellular Ca2+, and a blockade of intracellular Ca2+ increase inhibited the acid-induced increase in COX2 expression and PGE2 production. Knockdown of NOX5-S or NF-kappaB1 p50 by their small interfering RNA significantly inhibited acid-induced COX2 expression and PGE2 production in SEG1 cells. Acid treatment significantly decreased IkappaBalpha and increased luciferase activity when SEG1 cells were transfected with an NF-kappaB in vivo activation reporter plasmid, pNF-kappaB-Luc. In a novel Barrett's cell line overexpressing NOX5-S, IkappaBalpha was significantly reduced, and luciferase activity increased when these Barrett's cells were transfected with pNF-kappaB-Luc. Overexpression of NOX5-S in Barrett's cells significantly increased H2O2 production, COX2 expression, PGE2 production, and thymidine incorporation. The increase in thymidine incorporation occurring in NOX5-S-overexpressing Barrett's cells or induced by acid treatment in SEG1 EA cells was significantly decreased by COX2 inhibitors or small interfering RNA. We conclude that acid-induced COX2 expression and PGE2 production depend on an increase in cytosolic Ca2+ and sequential activation of NOX5-S and NF-kappaB in SEG1 cells. COX2-derived PGE2 production may contribute to NOX5-S-mediated cell proliferation in SEG1 cells. PMID- 17403675 TI - A conserved sequence immediately N-terminal to the Bateman domains in AMP activated protein kinase gamma subunits is required for the interaction with the beta subunits. AB - Mammalian AMP-activated protein kinase is a serine/threonine protein kinase that acts as a sensor of cellular energy status. AMP-activated protein kinase is a heterotrimer of three different subunits, i.e. alpha, beta, and gamma, with alpha being the catalytic subunit and beta and gamma having regulatory roles. Although several studies have defined different domains in alpha and beta involved in the interaction with the other subunits of the complex, little is known about the regions of the gamma subunits involved in these interactions. To study this, we have made sequential deletions from the N termini of the gamma subunit isoforms and studied the interactions with alpha and beta subunits, both by two-hybrid analysis and by co-immunoprecipitation. Our results suggest that a conserved region of 20-25 amino acids in gamma1, gamma2, and gamma3, immediately N-terminal to the Bateman domains, is required for the formation of a functional, active alphabetagamma complex. This region is required for the interaction with the beta subunits. The interaction between the alpha and gamma subunits does not require this region and occurs instead within the Bateman domains of the gamma subunit, although the alpha-gamma interaction does appear to stabilize the beta-gamma interaction. In addition, sequential deletions from the C termini of the gamma subunits indicate that deletion of any of the CBS (cystathionine beta-synthase) motifs prevents the formation of a functional complex with the alpha and beta subunits. PMID- 17403676 TI - Rin1 interacts with signal-transducing adaptor molecule (STAM) and mediates epidermal growth factor receptor trafficking and degradation. AB - Rin1, the prototype of a new family of multidomain Rab5 exchange factors, has been shown to play an important role in the endocytosis of the epidermal growth factor receptor (EGFR). Herein, we examined the role of Rin1 in the down regulation of EGFR following EGF stimulation. We observed that overexpression of Rin1 accelerates EGFR degradation in EGF-stimulated cells. In concordance, depletion of endogenous Rin1 by RNA interference resulted in a substantial reduction of EGFR degradation. We showed that Rin1 interacts with signal transducing adaptor molecule 2 (STAM2), a protein that associates with hepatocyte growth factor-regulated substrate and plays a key role in the endosomal sorting machinery. Green fluorescent protein (GFP)-Rin1 co-localizes with hemagglutinin (HA)-STAM2 and with endogenous hepatocyte growth factor-regulated substrate. Furthermore, wild type STAM2, but not a deletion mutant lacking the SH3 domain, co-immunoprecipitates with endogenous Rin1. This interaction is dependent on the proline-rich domain (PRD) of Rin1 as Rin1DeltaPRD, a mutant lacking the PRD, does not interact with STAM2. Moreover, EGFR degradation was not accelerated by expression of the Rin1DeltaPRD mutant. Together these results suggest that Rin1 regulates EGFR degradation in cooperation with STAM, defining a novel role for Rin1 in regulating endosomal trafficking. PMID- 17403677 TI - The major secreted cathepsin L1 protease of the liver fluke, Fasciola hepatica: a Leu-12 to Pro-12 replacement in the nonconserved C-terminal region of the prosegment prevents complete enzyme autoactivation and allows definition of the molecular events in prosegment removal. AB - A protease secreted by the parasitic helminth Fasciola hepatica, a 37-kDa procathepsin L1 (FheproCL1), autocatalytically processes and activates to its mature enzyme (FheCL1) over a wide pH range of 7.3 to 4.0, although activation is more rapid at low pH. Maturation initiates with cleavages of a small proportion of molecules within the central region of the prosegment, possibly by intramolecular events. However, activation to fully mature enzymes is achieved by a precise intermolecular cleavage at a Leu-12-Ser-11 downward arrowHis-10 sequence within the nonconserved C-terminal region of the prosegment. The importance of this cleavage site in enzyme activation was demonstrated using an active site variant FheproCL1Gly26 (Cys26 to Gly26) and a double variant FheproCL1Pro-12/Gly26 (Leu-12 to Pro-12), and although both of these variants cannot autocatalytically process, the former is susceptible to trans-processing at a Leu-12-Ser-11 downward arrowHis-10 sequence by pre-activated FheCL1, but the latter is not. Another F. hepatica secreted protease FheCL2, which, unlike FheCL1, can readily accept proline in the S2 subsite of its active site, can trans-process the double variant FheproCL1Pro-12/Gly26 by cleavage at the Pro-12 Ser-11 downward arrowHis-10 sequence. Furthermore, the autoactivation of a variant enzyme with a single replacement, FheproCL1Pro-12, was very slow but was increased 40-fold in the presence of FheCL2. These studies provide a molecular insight into the regulation of FheproCL1 autocatalysis. PMID- 17403678 TI - The Lipoxin A4 receptor is coupled to SHP-2 activation: implications for regulation of receptor tyrosine kinases. AB - Mesangial cell proliferation is pivotal to the pathology of glomerular injury in inflammation. We have previously reported that lipoxins, endogenously produced eicosanoids with anti-inflammatory and pro-resolution bioactions, can inhibit mesangial cell proliferation in response to several agents. This process is associated with elaborate receptor cross-talk involving modification receptor tyrosine kinase phosphorylation (McMahon, B., Mitchell, D., Shattock, R., Martin, F., Brady, H. R., and Godson, C. (2002) FASEB J. 16, 1817-1819). Here we demonstrate that the lipoxin A(4) (LXA(4)) receptor is coupled to activation and recruitment of the SHP-2 (SH2 domain-containing tyrosine phosphatase-2) within a lipid raft microdomain. Using site-directed mutagenesis of the cytosolic domain of the platelet-derived growth factor receptor beta (PDGFRbeta), we report that mutation of the sites for phosphatidylinositol 3-kinase (Tyr(740) and Tyr(751)) and SHP-2 (Tyr(763) and Tyr(1009)) recruitment specifically inhibit the effect of LXA(4) on the PDGFRbeta signaling; furthermore inhibition of SHP-2 expression with short interfering RNA constructs blocked the effect of LXA(4) on PDGFRbeta phosphorylation. We demonstrate that association of the PDGFRbeta with lipid raft microdomains renders it susceptible to LXA(4)-mediated dephosphorylation by possible reactivation of oxidatively inactivated SHP-2. These data further elaborate on the potential mechanisms underlying the anti-inflammatory, proresolution, and anti-fibrotic bioactions of lipoxins. PMID- 17403679 TI - Non-AUG translation initiation of a fungal RING finger repressor involved in photocarotenogenesis. AB - The RING finger protein CrgA acts as a negative regulator of light-induced carotene biosynthesis in the fungus Mucor circinelloides. Sequence analysis of the crgA coding region upstream of the first AUG codon revealed the existence of an additional non-canonical RING finger domain at the most N-terminal end of the protein. The newly identified RING finger domain is required for CrgA to regulate photocarotenogenesis, as deduced from site-directed mutagenesis experiments. The role of both RING finger domains in the stability of CrgA has been investigated in a yeast system. Wild type CrgA, but not the RING finger deleted forms, is highly unstable and is stabilized by inhibition of the proteasome function, which suggests that native CrgA is degraded by the proteasome and that active RING finger domains are required for proteasome-mediated CrgA degradation. To identify the translation start of CrgA, a mutational analysis of putative initiation codons in the 5' region of the crgA gene was accomplished. We demonstrated that a GUG codon located upstream of the first AUG is the sole initiator of CrgA translation. To our knowledge, this is the first report of a naturally occurring non-AUG start codon for a RING finger regulatory protein. A combination of suboptimal translation initiation and proteasome degradation may help to maintain the low cellular levels of CrgA observed in wild type cells, which is probably required for accurate regulation of photocarotenogenesis. PMID- 17403680 TI - Regulation of ADAMTS9 secretion and enzymatic activity by its propeptide. AB - ADAMTS9 is a secreted, cell-surface-binding metalloprotease that cleaves the proteoglycans versican and aggrecan. Unlike most precursor proteins, the ADAMTS9 zymogen (pro-ADAMTS9) is resistant to intracellular processing. Instead, pro ADAMTS9 is processed by furin at the cell surface. Here, we investigated the role of the ADAMTS9 propeptide in regulating its secretion and proteolytic activity. Removal of the propeptide abrogated secretion of the ADAMTS9 catalytic domain, and secretion was inefficiently restored by expression of the propeptide in trans. Substitution of Ala for Asn residues within each of three consensus N linked glycosylation sites in the propeptide abrogated ADAMTS9 secretion. Thus, the propeptide is an intramolecular chaperone whose glycosylation is critical for secretion of the mature enzyme. In addition to two previously identified furin processing sites (Arg74 downward arrow and Arg287 downward arrow) the ADAMTS9 propeptide was also furin-processed at Arg209. Substitution of Ala for Arg74, Arg209, and Arg287 resulted in secretion of an unprocessed zymogen. Unexpectedly, versican incubated with cells expressing this pro-ADAMTS9 was processed to a greater extent than when incubated with cells expressing wild-type, furin processable ADAMTS9. Moreover, cells and medium treated with the proprotein convertase inhibitor decanoyl-Arg-Val-Lys-Arg-chloromethyl ketone had greater versican-cleaving activity than untreated cells. Following furin processing of pro-ADAMTS9, propeptide fragments maintained a non-covalent association with the catalytic domain. Collectively, these observations suggest that, unlike other metalloproteases, furin processing of the ADAMTS9 propeptide reduces its catalytic activity. Thus, the propeptide is a key functional domain of ADAMTS9, mediating an unusual regulatory mechanism that may have evolved to ensure maximal activity of this protease at the cell surface. PMID- 17403681 TI - Interaction between HIV-1 Rev and integrase proteins: a basis for the development of anti-HIV peptides. AB - Human immunodeficiency virus 1 (HIV-1) Rev and integrase (IN) proteins are required within the nuclei of infected cells in the late and early phases of the viral replication cycle, respectively. Here we show using various biochemical methods, that these two proteins interact with each other in vitro and in vivo. Peptide mapping and fluorescence anisotropy showed that IN binds residues 1-30 and 49-74 of Rev. Following this observation, we identified two short Rev-derived peptides that inhibit the 3'-end processing and strand-transfer enzymatic activities of IN in vitro. The peptides bound IN in vitro, penetrated into cultured cells, and significantly inhibited HIV-1 in multinuclear activation of a galactosidase indicator (MAGI) and lymphoid cultured cells. Real time PCR analysis revealed that the inhibition of HIV-1 multiplication is due to inhibition of the catalytic activity of the viral IN. The present work describes novel anti-HIV-1 lead peptides that inhibit viral replication in cultured cells by blocking DNA integration in vivo. PMID- 17403682 TI - Interaction between familial amyotrophic lateral sclerosis (ALS)-linked SOD1 mutants and the dynein complex. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by progressive motor neuron death. More than 90 mutations in the copper-zinc superoxide dismutase (SOD1) gene cause a subset of familial ALS. Toxic properties have been proposed for the ALS-linked SOD1 mutants, but the nature of the toxicity has not been clearly specified. Cytoplasmic inclusion bodies containing mutant SOD1 and a number of other proteins are a pathological hallmark of mutant SOD1-mediated familial ALS, but whether such aggregates are toxic to motor neurons remains unclear. In this study, we identified a dynein subunit as a component of the mutant SOD1-containing high molecular weight complexes using proteomic techniques. We further demonstrated interaction and colocalization between dynein and mutant SOD1, but not normal SOD1, in cultured cells and also in G93A and G85R transgenic rodent tissues. Moreover, the interaction occurred early, prior to the onset of symptoms in the ALS animal models and increased over the disease progression. Motor neurons with long axons are particularly susceptible to defects in axonal transport. Our results demonstrate a direct "gain-of-interaction" between mutant SOD1 and dynein, which may provide insights into the mechanism by which mutant SOD1 could contribute to a defect in retrograde axonal transport or other dynein functions. The aberrant interaction is potentially critical to the formation of mutant SOD1 aggregates as well as the toxic cascades leading to motor neuron degeneration in ALS. PMID- 17403683 TI - MITF and PU.1 recruit p38 MAPK and NFATc1 to target genes during osteoclast differentiation. AB - Transcription factors NFATc1, PU.1, and MITF collaborate to regulate specific genes in response to colony-stimulating factor-1 (CSF-1) and receptor activator of NF-kappaB ligand (RANKL) signaling during osteoclast differentiation. However, molecular details concerning timing and mechanism of specific events remain ill defined. In bone marrow-derived precursors, CSF-1 alone promoted assembly of MITF PU.1 complexes at osteoclast target gene promoters like cathepsin K and acid 5 phosphatase without increasing gene expression. The combination of RANKL and CSF 1 concurrently increased the levels of MAPK-phosphorylated forms of MITF, p38 MAPK, and SWI/SNF chromatin-remodeling complexes bound to these target promoters and markedly increased expression of the genes. NFATc1 was subsequently recruited to complexes at the promoters during terminal stages of osteoclast differentiation. Genetic analysis of Mitf and Pu.1 in mouse models supported the critical interaction of these genes in osteoclast differentiation. The results define MITF and PU.1 as nuclear effectors that integrate CSF-1/RANKL signals during osteoclast differentiation to initiate expression of target genes, whereas a complex that includes NFATc1 may act to maintain target gene expression in differentiated cells. PMID- 17403684 TI - Post-translational methylation of high mobility group box 1 (HMGB1) causes its cytoplasmic localization in neutrophils. AB - High mobility group box 1 (HMGB1) protein plays multiple roles in transcription, replication, and cellular differentiation. HMGB1 is also secreted by activated monocytes and macrophages and passively released by necrotic or damaged cells, stimulating inflammation. HMGB1 is a novel antigen of anti-neutrophil cytoplasmic antibodies (ANCA) observed in the sera of patients with ulcerative colitis and autoimmune hepatitis, suggesting that HMGB1 is secreted from neutrophils to the extracellular milieu. However, the actual distribution of HMGB1 in the cytoplasm of neutrophils and the mechanisms responsible for it are obscure. Here we show that HMGB1 in neutrophils is post-translationally mono-methylated at Lys42. The methylation alters the conformation of HMGB1 and weakens its DNA binding activity, causing it to become largely distributed in the cytoplasm by passive diffusion out of the nucleus. Thus, post-translational methylation of HMGB1 causes its cytoplasmic localization in neutrophils. This novel pathway explains the distribution of nuclear HMGB1 to the cytoplasm and is important for understanding how neutrophils release HMGB1 to the extracellular milieu. PMID- 17403685 TI - Vitamin C is an essential antioxidant that enhances survival of oxidatively stressed human vascular endothelial cells in the presence of a vast molar excess of glutathione. AB - Cellular glutathione levels may exceed vitamin C levels by 10-fold, generating the question about the real antioxidant role that low intracellular concentrations of vitamin C can play in the presence of a vast molar excess of glutathione. We characterized the metabolism of vitamin C and its relationship with glutathione in primary cultures of human endothelial cells oxidatively challenged by treatment with hydrogen peroxide or with activated cells undergoing the respiratory burst, and analyzed the manner in which vitamin C interacts with glutathione to increase the antioxidant capacity of cells. Our data indicate that: (i) endothelial cells express transporters for reduced and oxidized vitamin C and accumulate ascorbic acid with participation of glutathione-dependent dehydroascorbic acid reductases, (ii) although increased intracellular levels of vitamin C or glutathione caused augmented resistance to oxidative stress, 10 times more glutathione than vitamin C was required, (iii) full antioxidant protection required the simultaneous presence of intracellular and extracellular vitamin C at concentrations normally found in vivo, and (iv) intracellular vitamin C cooperated in enhancing glutathione recovery after oxidative challenge thus providing cells with enhanced survival potential, while extracellular vitamin C was recycled through a mechanism involving the simultaneous neutralization of oxidant species. Therefore, in endothelial cells under oxidative challenge, vitamin C functions as an essential cellular antioxidant even in the presence of a vast molar excess of glutathione. PMID- 17403686 TI - Hypoxia-inducible expression of a natural cis-antisense transcript inhibits endothelial nitric-oxide synthase. AB - The destabilization of endothelial nitric-oxide synthase (eNOS) mRNA in hypoxic endothelial cells may be important in the etiology of vascular diseases, such as pulmonary hypertension. Recently, an overlapping antisense transcript to eNOS/NOS3 was implicated in the post-transcriptional regulation of eNOS. We demonstrate here that expression of sONE, also known as eNOS antisense (NOS3AS) or autophagy 9-like 2 (APG9L2), is robustly induced by hypoxia or functional deficiency of von Hippel-Lindau protein. sONE is also up-regulated in the aortas of hypoxic rats. In hypoxic endothelial cells, sONE expression negatively correlates with eNOS expression. Blocking the hypoxic induction of sONE by RNA interference attenuates the fall in both eNOS RNA and protein. We provide evidence that the induction of sONE primarily involves transcript stabilization rather than increased transcriptional activity and is von Hippel-Lindaubut not hypoxia-inducible factor 2alpha-dependent. We also demonstrate that sONE transcripts are enriched in the nucleus of normoxic cells and that hypoxia promotes an increase in the level of cytoplasmic and polyribosome-associated, sONE mRNA. The finding that eNOS expression can be regulated by an overlapping cis-antisense transcript in a stimulus-dependent fashion provides evidence that sense/antisense interactions may play a previously unappreciated role in vascular disease pathogenesis. PMID- 17403687 TI - Remote control of gene expression. AB - The elucidation of a growing number of species' genomes heralds an unprecedented opportunity to ascertain functional attributes of non-coding sequences. In particular, cis regulatory modules (CRMs) controlling gene expression constitute a rich treasure trove of data to be defined and experimentally validated. Such information will provide insight into cell lineage determination and differentiation and the genetic basis of heritable diseases as well as the development of novel tools for restricting the inactivation of genes to specific cell types or conditions. Historically, the study of CRMs and their individual transcription factor binding sites has been limited to proximal regions around gene loci. Two important by-products of the genomics revolution, artificial chromosome vectors and comparative genomics, have fueled efforts to define an increasing number of CRMs acting remotely to control gene expression. Such regulation from a distance has challenged our perspectives of gene expression control and perhaps the very definition of a gene. This review summarizes current approaches to characterize remote control of gene expression in transgenic mice and inherent limitations for accurately interpreting the essential nature of CRM activity. PMID- 17403688 TI - Insights into the mechanism of partial agonism: crystal structures of the peroxisome proliferator-activated receptor gamma ligand-binding domain in the complex with two enantiomeric ligands. AB - The peroxisome proliferator-activated receptors (PPARs) are transcriptional regulators of glucose and lipid metabolism. They are activated by natural ligands, such as fatty acids, and are also targets of synthetic antidiabetic and hypolipidemic drugs. By using cell-based reporter assays, we studied the transactivation activity of two enantiomeric ureidofibrate-like derivatives. In particular, we show that the R-enantiomer, (R)-1, is a full agonist of PPARgamma, whereas the S-enantiomer, (S)-1, is a less potent partial agonist. Most importantly, we report the x-ray crystal structures of the PPARgamma ligand binding domain complexed with the R- and the S-enantiomer, respectively. The analysis of the two crystal structures shows that the different degree of stabilization of the helix 12 induced by the ligand determines its behavior as full or partial agonist. Another crystal structure of the PPARgamma.(S)-1 complex, only differing in the soaking time of the ligand, is also presented. The comparison of the two structures of the complexes with the partial agonist reveals significant differences and is suggestive of the possible coexistence in solution of transcriptionally active and inactive forms of helix 12 in the presence of a partial agonist. Mutation analysis confirms the importance of Leu(465), Leu(469), and Ile(472) in the activation by (R)-1 and underscores the key role of Gln(286) in the PPARgamma activity. PMID- 17403689 TI - GSK-3beta acts upstream of Fyn kinase in regulation of nuclear export and degradation of NF-E2 related factor 2. AB - NF-E2-related factor 2 (Nrf2) regulates expression and coordinated induction of a battery of chemoprotective genes in response to oxidative and electrophilic stress. This leads to protection against oxidative stress and neoplastic diseases. Nuclear import and export of Nrf2 play a significant role in control of nuclear levels of Nrf2 and thus the expression of Nrf2 down-stream genes. Tyrosine kinase Fyn phosphorylates tyrosine 568 of Nrf2 that leads to the nuclear export of Nrf2. In this study, we investigated the upstream factor(s) in regulation of Fyn and Fyn-mediated nuclear export of Nrf2. The investigations shed light on a novel mechanism of Nrf2 regulation in response to oxidative stress. We demonstrate that GSK-3beta acts upstream of Fyn kinase in control of nuclear export of Nrf2. Chemical and short interfering RNA-mediated inhibition of GSK-3beta led to nuclear accumulation of Nrf2 and transcriptional activation of the Nrf2 downstream gene nqo1. Chemical and short interfering RNA inhibition of GSK-3beta and Fyn individually and in combination revealed that both kinases follow the same pathway to regulate nuclear export of Nrf2. We further demonstrate that hydrogen peroxide phosphorylates tyrosine 216 of GSK-3beta. This leads to activation of GSK-3beta. The activated GSK-3beta phosphorylates Fyn at threonine residue(s). Phosphorylated Fyn accumulates in the nucleus and phosphorylates Nrf2 at tyrosine 568. This leads to nuclear export, ubiquitination, and degradation of Nrf2. PMID- 17403690 TI - Protein kinase Cdelta is activated by Shiga toxin and regulates its transport. AB - Protein kinase C (PKC) isozymes regulate different vesicular trafficking steps in the recycling or degradative pathways. However, a possible role of these kinases in the retrograde pathway from endosomes to the Golgi complex has previously not been investigated. We report here the involvement of a specific PKC isozyme, PKCdelta, in the intracellular transport of the glycolipid-binding Shiga toxin (Stx), which utilizes the retrograde pathway to intoxicate cells. Upon binding to cells, Stx was shown to specifically activate PKCdelta and not PKCalpha. The involvement of PKCdelta and PKCalpha in the retrograde transport of Stx was then monitored biochemically and by immunofluorescence after inhibition or depletion of the isozymes. PKCdelta, but not PKCalpha, was shown to selectively regulate the endosome-to-Golgi transport of StxB. Upon inhibition or knockdown of PKCdelta, StxB molecules colocalized less with giantin and more with EEA1, indicating that the molecules were accumulated in endosomes, unable to reach the Golgi complex. The inhibition of Golgi transport of Stx was reflected by a strong reduction in the toxic effect, demonstrating that transport of Stx to the cytosol is dependent on PKCdelta activity. These results are in agreement with our previous data, which show that Stx is able to stimulate its own transport. PMID- 17403691 TI - NXF2 is involved in cytoplasmic mRNA dynamics through interactions with motor proteins. AB - Tap/NXF1, the founding member of the evolutionarily conserved NXF (Nuclear RNA export Factor) family of proteins, is required for the nuclear export of bulk poly(A)+ RNAs. In mice, three additional NXF family genes (NXF2, NXF3, NXF7) have been identified and characterized to date. Cumulative data suggest that NXF family members play roles, not only in nuclear mRNA export, but also in various aspects of post-transcriptional mRNA metabolism. In order to better understand the functional role of NXF2, we searched for its binding partners by yeast two hybrid screening and identified several cytoplasmic motor proteins, including KIF17. The interaction of NXF2 with KIF17, which was confirmed by GST pull-down and co-immunoprecipitation assays, is mediated by the N-terminal domain of NXF2, which is required for the punctate localization patterns in dendrites of primary neurons. We also found that the NXF2-containing dendritic granules, which were co localized with KIF17, mRNA and Staufen1, a known component of neuronal mRNA granules, moved bidirectionally along dendrites in a microtubule-dependent manner. These results suggest that NXF2, a nucleo-cytoplasmic mRNA transporter, plays additional roles in the cytoplasmic localization of mRNAs through interactions with cytoplasmic motor proteins. PMID- 17403692 TI - Bidirectional transcription is an inherent feature of Giardia lamblia promoters and contributes to an abundance of sterile antisense transcripts throughout the genome. AB - A prominent feature of transcription in Giardia lamblia is the abundant production of sterile antisense transcripts (Elmendorf et al. The abundance of sterile transcripts in Giardia lamblia. Nucleic Acids., 29, 4674-4683). Here, we use a computational biology analysis of SAGE data to assess the abundance and distribution of sense and antisense messages in the parasite genome. Sterile antisense transcripts are produced at approximately 50% of loci with detectable transcription, yet their abundance at a given locus does not correlate to the abundance of the complementary sense transcripts at that locus or to transcription levels at neighboring loci. These data suggest that sterile antisense transcripts are not simply a local effect of open chromatin structure. Using 5'RACE, we demonstrate that Giardia promoters are a source of antisense transcripts through bidirectional transcription, producing both downstream coding sense and upstream sterile antisense transcripts. We use a dual reporter system to explore roles of specific promoter elements in this bidirectional initiation of transcription and suggest that the degenerate AT-rich nature of TATA and Inr elements in Giardia permits them to function interchangeably. The phenomenon of bidirectional transcription in G. lamblia gives us insight into the interaction between transcriptional machinery and promoter elements, and may be the prominent source of the abundant antisense transcription in this parasite. PMID- 17403693 TI - Mapping biomedical concepts onto the human genome by mining literature on chromosomal aberrations. AB - Biomedical literature provides a rich but unstructured source of associations between chromosomal regions and biomedical concepts. By mining MEDLINE abstracts, we annotate the human genome at the level of cytogenetic bands. Our method creates a set of chromosomal aberration maps that associate cytogenetic bands to biomedical concepts from a variety of controlled vocabularies, including disease, dysmorphology, anatomy, development and Gene Ontology branches. The association between a band (e.g. 4p16.3) and a concept (e.g. microcephaly) is assessed by the statistical overrepresentation of this concept in the abstracts relating to this band. Our method is validated using existing genome annotation resources and known chromosomal aberration maps and is further illustrated through a case study on heart disease. Our chromosomal aberration maps provide diagnostics support to clinical geneticists, aid cytogeneticists to interpret and report cytogenetic findings and support researchers interested in human gene function. The method is available as a web application, aBandApart, at http://www.esat.kuleuven.be/abandapart/. PMID- 17403694 TI - Nitric oxide controls nuclear export of APE1/Ref-1 through S-nitrosation of cysteines 93 and 310. AB - Apurinic/apyrimidinic endonuclease 1/redox effector factor-1 (APE1/Ref-1, abbreviated as APE1) is a molecule with dual functions in DNA repair and redox regulation of transcription factors. Accumulated work has shown that the biological activities of APE1 are sensitive to oxidative stress; however, whether APE1 functions can be regulated by nitrosative stress remains unknown. In this investigation, we found that S-nitrosoglutathion (GSNO), a nitric oxide donor and also an S-nitrosating agent, effectively stimulated nuclear export of APE1 in a CRM1-independent manner. This nuclear-cytoplasmic translocation was dependent on S-nitrosation modification of APE1, as simultaneous mutation of S-nitrosation target sites Cys93 and Cys310 completely abrogated the cytoplasmic redistribution. The translocation process was reversal and specific, as it could be reversed by reductive reagents, but could not be mimicked by H2O2. In structure, the region aa.64-80 and the beta-strand aa.311-316 in proximity to Cys93 and Cys310 were important for GSNO-induced APE1 relocalization. In addition, a defect of importin-mediated nuclear import pathway was found in the NO-insulted cells, and p50 and HDAC2 were identified as APE1 nuclear export inhibitory proteins. Together, this study may provide a novel molecular mechanism, which links nitrosative stress to APE1-associated physiological and pathological processes. PMID- 17403695 TI - Renal hypoplasia without optic coloboma associated with PAX2 gene deletion. PMID- 17403696 TI - Expression of retinoic acid-inducible gene-I in lupus nephritis. PMID- 17403697 TI - Fibrin sheath removal from central venous catheters: an internal snare manoeuvre. AB - BACKGROUND: Dysfunction of haemodialysis catheters is most commonly due to a narrowing of the catheter lumen and/or formation of a fibrin sheath around the catheter tip. Reported methods for restoring patency of the catheter lumen include passage of a J-tipped guide wire, passage of a biopsy brush through the catheter, or infusion of a thrombolytic agent into the catheter. While these methods are often effective, they suffer from several limitations. We present a minimally invasive technique to remove thrombi and debris from within the lumen of a partially thrombosed haemodialysis catheter while simultaneously stripping the fibrous sheath. METHODS: A 0.089 cm nitinol wire is bent to create a loop, which is then inserted via the catheters. Upon exiting the lumen of the catheters, the nitinol wire forces a snare open, which disrupts the fibrin sheath and catches intraluminal thrombi and debris. The technique requires no anaesthesia or recovery time. RESULTS: Initial clinical success in our series was achieved in all patients (7/7) as evidenced by restoration of target flow rates on subsequent haemodialysis. None of the patients experienced any complications as a result of the procedure. The catheter 2-, 4-, and 6-week primary success rates were 100% (8/8), 100% (8/8), and 100% (8/8) respectively with a mean duration of 17.1 weeks (range 8-40 weeks). CONCLUSIONS: The internal snare technique is an effective, inexpensive and minimally invasive approach to restoring patency to failed central venous access catheters. PMID- 17403698 TI - Expression of gremlin, a bone morphogenetic protein antagonist, in glomerular crescents of pauci-immune glomerulonephritis. AB - BACKGROUND: Recent evidence in vitro and in vivo suggests that gremlin, a bone morphogenetic protein antagonist, is participating in tubular epithelial mesenchymal transition (EMT) in diabetic nephropathy as a downstream mediator of TGF-beta. Since EMT also occurs in parietal epithelial glomerular cells (PECs) leading to crescent formation, we hypothesized that gremlin could participate in this process. With this aim we studied its expression in 30 renal biopsies of patients with pauci-immune crescentic nephritis. METHODS: Gremlin was detected by in situ hybridization (ISH) and immunohistochemistry (IMH) and TGF-beta by ISH and Smads by southwestern histochemistry (SWH). Phosphorylated Smad2, CTGF, BMP 7, PCNA, alpha-SMA, synaptopodin, CD-68, and phenotypic markers of PECs (cytokeratin, E-cadherin), were detected by IMH. In cultured human monocytes, gremlin and CTGF induction by TGF-beta was studied by western blot. RESULTS: We observed strong expression of gremlin mRNA and protein in cellular and fibrocellular crescents corresponding to proliferating PECs and monocytes, in co localization with TGF-beta. A marked over-expression of gremlin was also observed in tubular and infiltrating interstitial cells, correlating with tubulointerstitial fibrosis (r=0.59; P<0.01). A nuclear Smad activation in the same tubular cells, that are expressing TGF-beta and gremlin, was detected. In human cultured monocytes, TGF-beta induced gremlin production while CTGF expression was not detected. CONCLUSION: We postulate that gremlin may play a role in the fibrous process in crescentic nephritis, both in glomerular crescentic and tubular epithelial cells. The co-localization of gremlin and TGF beta expression found in glomeruli and tubular cells suggest that gremlin may be important in mediating some of the pathological effects of TGF-beta. PMID- 17403699 TI - How a skillful (correction of skilful) and motivated urinary sediment examination can save the kidneys. PMID- 17403700 TI - In vitro activity of daptomycin and vancomycin lock solutions on staphylococcal biofilms in a central venous catheter model. AB - BACKGROUND: Catheter lock solutions are used for prevention and management of catheter-related bloodstream infections. We investigated the activity of daptomycin and vancomycin lock solutions against Staphylococcus aureus and Staphylococcus epidermidis in an in vitro central venous catheter (CVC) model. METHODS: Biofilm-producing reference strains of S. aureus and S. epidermidis were evaluated. After 24 h of bacterial growth in a CVC model, daptomycin and vancomycin bactericidal activity (+/- preservative-containing heparin sodium) were separately evaluated as a lock solution using 0.5, 1 and 35 mg/ml. Calcium carbonate (50 mg/l) was added to all lock solutions containing daptomycin. Each CVC was drained, flushed and sonicated at 72 h to assess CFU/ml. RESULTS: After 72 h of exposure in the catheter lock solutions, daptomycin and vancomycin at 0.5, 1 and 5 mg/ml demonstrated bactericidal activity (>3.0 log10 CFU/ml) against S. aureus and S. epidermidis (P 95% of the individuals. METHODS: A total of 60 ASA I patients were randomly allocated to five different alfentanil dose groups (0, 15, 30, 45, or 60 microg kg-1). A blinded dose of alfentanil followed by thiopentone 4 mg kg-1 and rocuronium 1 mg kg-1 was administered in rapid succession, and tracheal intubation was attempted 40 s thereafter. The relationship between the alfentanil dose and the probability of optimal intubation conditions was determined by non-linear logistic regression analysis. Blood pressure (BP) changes were recorded continuously using an intra arterial catheter. RESULTS: The success rate of optimal intubation conditions increased with increasing doses of alfentanil. The alfentanil dose needed to obtain optimal intubation conditions in >95% of the patients was 36.4 (CI 33.4 39.4) microg kg-1. In 12 patients, the systolic BP declined to <90 mm Hg during the 3 min immediately after intubation. CONCLUSION: Adding 36-40 microg kg-1 alfentanil to a regimen of thiopentone and rocuronium during RSI of anaesthesia may significantly increase the success rate of optimal intubation conditions. Significant hypotension requiring vasopressor treatment may occur. PMID- 17403708 TI - Reversal of resistance in microorganisms by help of non-antibiotics. AB - Intracellular efflux pumps have been largely the research focus in multidrug resistant (MDR) Gram-positive and Gram-negative microorganisms and parasites including cancers. However, drug efflux mechanisms other than pumps per se have been observed, supported by the effects of isomeric, non-antibiotic depressant (DPR), phenothiazines and thixenes, and antidepressant (ADPR) phenylpiperidine neurotropic drugs, alone or in combination with classical antimicrobials on MDR Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus pyogenes and Streptococcus pneumoniae. Of the non-antibiotics we investigated, the DPR l-thioridazine, trans-clopenthixol and isomers of phenylpiperidines NNC 20-4962 (isomer of femoxetine) and NNC 20-7052 (isomer of paroxetine) were potent antimicrobials with the least neurotropic activity, pointing to a possible general isomeric structure-activity relationship. These compounds may be regarded as new efflux inhibitors. Moreover, these isomers have considerably reduced, in some cases absent, neurotropism and reduced mammalian toxicity. This may alleviate concerns about adverse effects and therapeutic safety for infected patients in life-threatening situations where the non antibiotic dosage would be in the lower, non-chronic dosage ranges generally prescribed for individuals with mild mental health problems. The results point to the prokaryotic and eukaryotic microorganisms' phospholipid/protein domain involvement of the cationic, amphiphilic, non-antibiotic DPR and ADPR, with the phospholipids being the initial sites attracting and concentrating the neurotropes to induce a form of suspended animation, followed by gross changes of cell wall and membrane structure, and thereby potentiating their destructive or immobilizing effects on various as yet only hinted at resistance and efflux mechanisms. Combination of appropriate isomeric non-antibiotic DPR and ADPR of low neurotropism and toxicity with conventional and classical antimicrobials promises early, new therapeutic strategies salutary against microbial resistance, resistance development, pathogenicity and virulence. PMID- 17403709 TI - Molecular characterization of the gene encoding a new AmpC beta-lactamase in Acinetobacter baylyi. AB - OBJECTIVES: The main objective of the present study was to demonstrate the presence of a beta-lactamase ampC gene in the chromosome of the non-pathogenic bacterium Acinetobacter baylyi ADP1. METHODS: beta-Lactam MICs were determined by Etest. The ampC gene was amplified by PCR, with specific oligonucleotides, then cloned into pBGS18 and pAT-RA plasmids and transformed into Escherichia coli TG1 and parental A. baylyi as hosts. The gene was sequenced and analysed. The AmpC protein was expressed, purified by affinity chromatography and the kinetic parameters determined. RESULTS: An ampC gene was amplified from the ADP1 genome. Sequencing of the gene showed typical SVSK and KTG domains and the typical YXN Class C motif. The amplified gene showed significant identity (48.5% to 49.3%) with the AmpC enzymes of Acinetobacter baumannii and AG3 strains, which have recently been renamed ADC-1 to ADC-7. MIC analysis revealed a cephalosporinase profile for the E. coli TG1 clone as well as for the parental A. baylyi strain that overexpressed the ampC gene cloned under the control of an external promoter. Analysis of kinetic parameters of the purified enzyme showed higher catalytic efficiency for cefalotin than for ampicillin. CONCLUSIONS: This study represents the first report of an AmpC beta-lactamase in A. baylyi, which was shown by biochemical and microbiological experiments to have a typical cephalosporinase profile. The presence of the respective gene in the chromosome of A. baylyi ADP1 suggested that this ampC gene is the naturally occurring cephalosporinase in this species, as previously reported for other Acinetobacter spp. We tentatively named the enzyme ADC-8. PMID- 17403710 TI - Prevalence and complications of uveitis in juvenile idiopathic arthritis in a population-based nation-wide study in Germany: suggested modification of the current screening guidelines. AB - OBJECTIVES: To analyse the prevalence and complications of uveitis and their predictors in a large cohort of patients with juvenile idiopathic arthritis (JIA). METHODS: Data of 3271 JIA patients as classified by International League of Associations for Rheumatology (ILAR) criteria included in a national database during 1 yr were analysed. RESULTS: Uveitis prevalence was 12% of all JIA patients. The most frequent were oligoarthritis extended (25%) and persistent (16%). JIA patients with uveitis were significantly younger at onset of arthritis (3.8 vs 7.0 yrs) or ANA-positive (86% vs 42%) than the patients without uveitis. Predictors of uveitis included age at onset (P= 0.03) and ANA-positivity (P< 0.01) besides the presence of a certain JIA subgroup (P= 0.04). Uveitis was clinically silent in 75% of the oligoarthritis but in none of the enthesitis related arthritis patients. The median onset of uveitis was 5.5 months after arthritis manifestation. In 73%, 77% and 90%, uveitis developed within 1, 2 and 4 yrs after arthritis, respectively. Anterior uveitis was the most common anatomic type of uveitis (83%). Uveitis complications at mean follow-up of 5.6 yrs were common (56%), and predictors for complications included presence of complications at first visit (P< 0.001) and uveitis manifestation before arthritis (P= 0.001), but not ANA positivity. CONCLUSIONS: The JIA subgroups markedly differ with respect to the prevalence and course of associated uveitis. Ophthalmological screening should be initiated early after arthritis onset and the intervals be related to the JIA subgroup. A modification of the current screening guidelines is suggested. PMID- 17403711 TI - Vasculopathy and arterial stenotic lesions in the antiphospholipid syndrome. AB - The antiphospholipid (Hughes) syndrome (APS) is characterized by recurrent arterial or venous thromboembolism, or pregnancy loss, in association with antiphospholipid antibodies. These antibodies may be associated with premature or accelerated atherosclerosis and emerging evidence supports the concept of a vasculopathy in the APS that may lead to arterial stenotic lesions, possibly contributing to vascular occlusions and pregnancy morbidity. PMID- 17403712 TI - Anti-TNF therapy in the management of Behcet's disease--review and basis for recommendations. PMID- 17403713 TI - Problems with use of composite end points in cardiovascular trials: systematic review of randomised controlled trials. AB - OBJECTIVE: To explore the extent to which components of composite end points in randomised controlled trials vary in importance to patients, the frequency of events in the more and less important components, and the extent of variability in the relative risk reductions across components. DESIGN: Systematic review of randomised controlled trials. DATA SOURCES: Cardiovascular randomised controlled trials published in the Lancet, Annals of Internal Medicine, Circulation, European Heart Journal, JAMA, and New England Journal of Medicine, from 1 January 2002 to 30 June 2003. Component end points of composite end points were categorised according to importance to patients as fatal, critical, major, moderate, or minor. RESULTS: Of 114 identified randomised controlled trials that included a composite end point of importance to patients, 68% (n=77) reported complete component data for the primary composite end point; almost all (98%; n=112) primary composite end points included a fatal end point. Of 84 composite end points for which component data were available, 54% (n=45) showed large or moderate gradients in both importance to patients and magnitude of effect across components. When analysed by categories of importance to patients, the most important components were associated with lower event rates in the control group (medians of 3.3-3.7% for fatal, critical, and major outcomes; 12.3% for moderate outcomes; and 8.0% for minor outcomes). Components of greater importance to patients were associated with smaller treatment effects than less important ones (relative risk reduction of 8% for death and 33% for components of minor importance to patients). CONCLUSION: The use of composite end points in cardiovascular trials is frequently complicated by large gradients in importance to patients and in magnitude of the effect of treatment across component end points. Higher event rates and larger treatment effects associated with less important components may result in misleading impressions of the impact of treatment. PMID- 17403715 TI - Contribution of the putative genetic factors and ANKH gene polymorphisms to variation of circulating calciotropic molecules, PTH and BGP. AB - It is well known that regulation of calcium homeostasis in bone remodeling is one of the most crucial factors for maintaining healthy bones. Parathyroid hormone (PTH) is probably the most important hormone that participates in the bone remodeling process. Another important biochemical factor governing bone metabolism is osteocalcin (BGP). Although the physiological functions of both of these factors are well known, there is still very little known regarding their specific genetic determination and in particular, the specific genes that may regulate the circulating concentrations of these substances. In the present study, we examined whether nine single nucleotide polymorphisms (SNPs) in the human homologue of the mouse progressive ankylosis gene (ANKH)-one of the key genetic factors involved in bone mineralization-can be associated with PTH and BGP levels in apparently healthy human populations. The study sample comprised 244 nuclear families (840 individuals). After adjustment of BGP and PTH for the significant covariates (sex, age and BMI), the contribution of the putative genetic effects was statistically significant (P < 0.001) for both biochemical factors: 45.27 +/- 10.8% for PTH and 30.19 +/- 12.6% for BGP. Application of transmission disequilibrium tests (TDTs) revealed a significant association (P < 0.05) between PTH and two SNPs: rs39968 and rs875525. However, the association became particularly significant for four TDTs (P-values ranging from 0.0025 to 0.0008) when the association with the haplotypes generated from the above SNP was tested. This association remained significant even after correction for multiple testing with a false discovery rate of 0.05. PMID- 17403714 TI - Impaired complex III assembly associated with BCS1L gene mutations in isolated mitochondrial encephalopathy. AB - We investigated two unrelated children with an isolated defect of mitochondrial complex III activity. The clinical picture was characterized by a progressive encephalopathy featuring early-onset developmental delay, spasticity, seizures, lactic acidosis, brain atrophy and MRI signal changes in the basal ganglia. Both children were compound heterozygotes for novel mutations in the human bc1 synthesis like (BCS1L) gene, which encodes an AAA mitochondrial protein putatively involved in both iron homeostasis and complex III assembly. The pathogenic role of the mutations was confirmed by complementation assays, using a DeltaBcs1 strain of Saccharomyces cerevisiae. By investigating complex III assembly and the structural features of the BCS1L gene product in skeletal muscle, cultured fibroblasts and lymphoblastoid cell lines from our patients, we have demonstrated, for the first time in a mammalian system, that a major function of BCS1L is to promote the maturation of complex III and, more specifically, the incorporation of the Rieske iron-sulfur protein into the nascent complex. Defective BCS1L leads to the formation of a catalytically inactive, structurally unstable complex III. We have also shown that BCS1L is contained within a high-molecular-weight supramolecular complex which is clearly distinct from complex III intermediates. PMID- 17403716 TI - COL10A1 nonsense and frame-shift mutations have a gain-of-function effect on the growth plate in human and mouse metaphyseal chondrodysplasia type Schmid. AB - Missense, nonsense and frame-shift mutations in the collagen X gene (COL10A1) result in metaphyseal chondrodysplasia type Schmid (MCDS). Complete degradation of mutant COL10A1 mRNA by nonsense-mediated decay in human MCDS cartilage implicates haploinsufficiency in the pathogenesis for nonsense mutations in vivo. However, the mechanism is unclear in situations where the mutant mRNA persist. We show that nonsense/frame-shift mutations can elicit a gain-of-function effect, affecting chondrocyte differentiation in the growth plate. In an MCDS proband, heterozygous for a p.Y663X nonsense mutation, the growth plate cartilage contained 64% wild-type and 36% mutant mRNA and the hypertrophic zone was disorganized and expanded. The in vitro translated mutant collagen X chains, which are truncated, were misfolded, unable to assemble into trimers and interfered with the assembly of normal alpha1(X) chains into trimers. Unlike Col10a1 null mutants, transgenic mice (FCdel) bearing the mouse equivalent of a human MCDS p.P620fsX621 mutation, displayed typical characteristics of MCDS with disproportionate shortening of limbs and early onset coxa vara. In FCdel mice, the degree of expansion of the hypertrophic zones was transgene-dosage dependent, being most severe in mice homozygous for the transgene. Chondrocytes in the lower region of the expanded hypertrophic zone expressed markers uncharacteristic of hypertrophic chondrocytes, indicating that differentiation was disrupted. Misfolded FCdel alpha1(X) chains were retained within the endoplasmic reticulum of hypertrophic chondrocytes, activating the unfolded protein response. Our findings provide strong in vivo evidence for a gain-of-function effect that is linked to the activation of endoplasmic reticulum-stress response and altered chondrocyte differentiation, as a possible molecular pathogenesis for MCDS. PMID- 17403717 TI - HEM dysplasia and ichthyosis are likely laminopathies and not due to 3beta hydroxysterol Delta14-reductase deficiency. AB - Mutations of the lamin B receptor (LBR) have been shown to cause HEM dysplasia in humans and ichthyosis in mice. LBR is a bifunctional protein with both a lamin B binding and a sterol Delta(14)-reductase domain. It previously has been proposed that LBR is the primary sterol Delta(14)-reductase and that HEM dysplasia and ichthyosis are inborn errors of cholesterol synthesis. However, DHCR14 also encodes a sterol Delta(14)-reductase and could provide enzymatic redundancy with respect to cholesterol synthesis. To test the hypothesis that LBR and DHCR14 both function as sterol Delta(14)-reductases, we obtained ichthyosis mice (Lbr(-/-)) and disrupted Dhcr14. Heterozygous Lbr and Dhcr14 mice were intercrossed to test for a digenic phenotype. Lbr(-/-), Dhcr14(Delta4-7/Delta4-7) and Lbr(+/ ):Dhcr14(Delta4-7/Delta4-7) mutant mice have distinct physical and biochemical phenotypes. Dhcr14(Delta4-7/Delta4-7) mice are essentially normal, whereas Lbr(+/ ):Dhcr14(Delta4-7/Delta4-7) mice are growth retarded and neurologically abnormal. Neither of these mutants resembles the ichthyosis mouse and biochemically, no sterol abnormalities were detected in either liver or kidney tissue. In contrast, relatively small transient elevations of Delta(14)-sterols were observed in Lbr( /-) and Dhcr14(Delta4-7/Delta4-7) brain tissue, and marked elevations were seen in Lbr(+/-):Dhcr14(Delta4-7/Delta4-7) brain. Pathological evaluation demonstrated vacuolation and swelling of the myelin sheaths in the spinal cord of Lbr(+/ ):Dhcr14(Delta4-7/Delta4-7) mice consistent with a demyelinating process. This was not observed in either Lbr(-/-) or Dhcr14 (Delta4-7/Delta4-7) mice. Our data support the conclusions that LBR and DHCR14 provide substantial enzymatic redundancy with respect to cholesterol synthesis and that HEM dysplasia and ichthyosis are laminopathies rather than inborn errors of cholesterol synthesis. PMID- 17403718 TI - Modulation of nucleosome dynamics in Huntington's disease. AB - Transcriptional dysregulation and aberrant chromatin remodeling are central features in the pathology of Huntington's disease (HD). In order to more fully characterize these pathogenic events, an assessment of histone profiles and associated gene changes were performed in transgenic N171-82Q (82Q) and R6/2 HD mice. Analyses revealed significant chromatin modification, resulting in reduced histone acetylation with concomitant increased histone methylation, consistent with findings observed in HD patients. While there are no known interventions that ameliorate or arrest HD progression, DNA/RNA-binding anthracyclines may provide significant therapeutic potential by correcting pathological nucleosome changes and realigning transcription. Two such anthracyclines, chromomycin and mithramycin, improved altered nucleosome homeostasis in HD mice, normalizing the chromatin pattern. There was a significant shift in the balance between methylation and acetylation in treated HD mice to that found in wild-type mice, resulting in greater acetylation of histone H3 at lysine 9 and promoting gene transcription. Gene expression profiling in anthracycline-treated HD mice showed molecular changes that correlate with disease correction, such that a subset of downregulated genes were upregulated with anthracycline treatment. Improved nucleosomal dynamics were concurrent with a significant improvement in the behavioral and neuropathological phenotype observed in HD mice. These data show the ability of anthracycline compounds to rebalance epigenetic histone modification and, as such, may provide the rationale for the design of human clinical trials in HD patients. PMID- 17403719 TI - Increase in serum adiponectin concentration in patients with heart failure and cachexia: relationship with leptin, other cytokines, and B-type natriuretic peptide. AB - AIMS: Adiponectin is a fat-derived hormone involved in the regulation of metabolism. Adiponectin concentration is inversely related to body weight and, in animals, causes weight loss. We, therefore, measured adiponectin concentration in patients with heart failure (HF) and cachexia. METHODS AND RESULTS: Serum adiponectin concentrations were measured in three groups of patients with coronary artery disease (CAD): (i) HF, reduced left ventricular systolic function, and cachexia (n = 10); (ii) HF, reduced systolic function but no cachexia (n = 20); (iii) HF-controls-patients with CAD, no HF, and preserved systolic function (n = 10); and in a healthy control group (n = 7). Patients with HF and cachexia had higher concentrations of adiponectin [23.8 (10.2-37.2) microg/mL] than all other groups: HF-no cachexia 8.1 (0.5-16.6) microg/mL; CAD controls 7.1 (0.4-13.5) microg/mL; and healthy controls 8.7 (2.5-16.8) microg/mL) (P < 0.05 for each comparison). Adiponectin correlated negatively with body mass index, percentage of body fat, waist circumference and insulin resistance, and positively with B-type natriuretic peptide (BNP) and tumour necrosis factor alpha. CONCLUSION: Cachexia in HF is associated with an increase in adiponectin concentration. This may represent preservation of the physiological response to change in body fat but might also suggest that adiponectin plays a role in the pathogenesis of cachexia. The correlation between BNP and adiponectin also raises the possibility that the former might increase the secretion of the latter. PMID- 17403720 TI - Waist circumference and waist-to-hip ratio as predictors of cardiovascular events: meta-regression analysis of prospective studies. AB - AIMS: The objectives of this study were to determine the association of waist circumference (WC) and waist-to-hip ratio (WHR) with the risk of incident cardiovascular disease (CVD) events and to determine whether the strength of association of WC and WHR with CVD risk is different. METHODS AND RESULTS: This meta-regression analysis used a search strategy of keywords and MeSH terms to identify prospective cohort studies and randomized clinical trials of CVD risk and abdominal obesity from the Medline, Embase, and Cochrane databases. Fifteen articles (n = 258 114 participants, 4355 CVD events) reporting CVD risk by categorical and continuous measures of WC and WHR were included. For a 1 cm increase in WC, the relative risk (RR) of a CVD event increased by 2% (95% CI: 1 3%) overall after adjusting for age, cohort year, or treatment. For a 0.01 U increase in WHR, the RR increased by 5% (95% CI: 4-7%). These results were consistent in men and women. Overall risk estimates comparing the extreme quantiles of each measure suggested that WHR was more strongly associated with CVD than that for WC (WHR: RR = 1.95, 95% CI: 1.55-2.44; WC: RR = 1.63, 95% CI: 1.31-2.04), although this difference was not significant. The strength of association for each measure was similar in men and women. CONCLUSION: WHR and WC are significantly associated with the risk of incident CVD events. These simple measures of abdominal obesity should be incorporated into CVD risk assessments. PMID- 17403721 TI - Synergistic relationship between hyperglycaemia and inflammation with respect to clinical outcomes in non-ST-elevation acute coronary syndromes: analyses from OPUS-TIMI 16 and TACTICS-TIMI 18. AB - AIMS: To investigate the relationship between diabetes and inflammation and the potentially synergistic relationship between hyperglycaemia and inflammation on clinical outcomes in non ST-elevation ACS. METHODS AND RESULTS: The principal analysis was conducted in 2200 patients in OPUS-TIMI 16 with C-reactive protein data available and then validated in the invasive arm of TACTICS-TIMI 18 (n = 929). In addition, two further inflammatory markers [monocyte chemoattractant protein-1 (MCP-1) and von Willebrand factor (vWF)] were assessed in OPUS-TIMI 16. Diabetic patients had higher C-reactive protein and MCP-1 levels vs. non-diabetic patients in OPUS-TIMI 16 (9 vs. 7.8 mg/L, P = 0.002, and 190.6 vs. 170.8 pg/mL, P = 0.04, respectively), higher C-reactive protein levels in TACTICS-TIMI 18 (6.6 vs. 5.2 mg/L, P = 0.0005), and as expected higher glucose levels in both trials. Stratifying by the median C-reactive protein and diabetes in OPUS-TIMI 16, diabetic patients with C-reactive protein greater than or equal to the median were the highest risk group vs. non-diabetic patients with C-reactive protein less than the median (adjusted HR 1.63, 95% CI 1.20-2.23, P = 0.002). Directionally, similar findings were observed for MCP-1 and vWF in OPUS-TIMI 16 and for C-reactive protein in TACTICS-TIMI 18. After adjustment for diabetes, the risk associated with a 1 mmol/L increase in glucose was greater among those with a C-reactive protein greater than or equal to the median (HR 1.07, 95% CI 1.03 1.11) vs. those with a C-reactive protein less than the median (HR 1.02, 95% CI 0.97-1.06). After multivariable adjustment, the synergistic relationship between glucose and C-reactive protein and clinical outcomes remained statistically significant (P = 0.01). A similar pattern was observed in TACTICS-TIMI 18. CONCLUSION: Among ACS patients, diabetes was associated with both greater inflammation and higher glucose levels and patients with both hyperglycaemia and inflammation had worse outcomes. Better control of both inflammation and hyperglycaemia should be assessed in future ACS trials as a means to reduce the cardiovascular risk among diabetics. PMID- 17403722 TI - The clinical expert: an empiric oddity. PMID- 17403723 TI - Medicine for dental students. PMID- 17403724 TI - Periodontal risk. PMID- 17403725 TI - Medicine and GPRs. PMID- 17403726 TI - More about medicine. PMID- 17403727 TI - Ethical moment questioned. PMID- 17403728 TI - Ethical moment hailed. PMID- 17403729 TI - Craniofacial pain. PMID- 17403730 TI - More about references. PMID- 17403731 TI - The role of references. PMID- 17403732 TI - Personnel shortages. PMID- 17403734 TI - Folic acid supplements taken early in pregnancy reduce facial cleft risk. PMID- 17403736 TI - The evidence base for the efficacy of antibiotic prophylaxis in dental practice. AB - INTRODUCTION: People with various medical conditions and devices are suggested candidates for receiving antibiotic prophylaxis before undergoing dental procedures. This practice is controversial, however, owing to the lack of proof of efficacy. The authors conducted a qualitative, systematic review to determine the level of evidence for this practice and whether antibiotic prophylaxis prevents distant site infections in these patients. METHODS: The authors selected eight groups of patients with specific medical conditions and devices who often are given antibiotic prophylaxis before undergoing invasive dental procedures. The conditions and devices were cardiac-native heart valve disease, prosthetic heart valves and pacemakers; hip, knee and shoulder prosthetic joints; renal dialysis shunts; cerebrospinal fluid shunts; vascular grafts; immunosuppression secondary to cancer and cancer chemotherapy; systemic lupus erythematosus; and insulin-dependent (type 1) diabetes mellitus. The authors thoroughly searched the literature for the years 1966 through 2005 for references indicating some level of support for this practice and graded each publication on the basis of level of evidence. RESULTS: The authors found formal recommendations in favor of antibiotic prophylaxis for only three of the eight medical conditions: native heart disease, prosthetic heart valves and prosthetic joints. They found no prospective randomized clinical trials and only one clinical study of antibiotic prophylaxis. Only one systematic review and two case series provided weak, if any, support for antibiotic prophylaxis in patients with cardiac conditions. The authors found little or no evidence to support this practice or to demonstrate that it prevents distant site infections for any of these eight groups of patients. CONCLUSIONS: No definitive, scientific basis exists for the use of prophylactic antibiotics before dental procedures for these eight groups of patients. PMID- 17403737 TI - The pathophysiology, medical management and dental implications of adult attention-deficit/hyperactivity disorder. AB - BACKGROUND: Few published reports in the dental literature have focused on adult attention-deficit/hyperactivity disorder (ADHD) and its dental implications. TYPES OF STUDIES REVIEWED: The authors conducted a MEDLINE search for the period 2000 through 2005 using the terms "adult" and "attention-deficit" to define ADHD's pathology, medical treatment and dental implications. RESULTS: ADHD is a developmental condition that affects slightly more than 4 percent of the adult U.S. population. Its symptoms include inattention, hyperactivity and impulsivity that can cause personal, social, occupational and leisure-time dysfunction. Medications used to treat the disorder include stimulants, selective noradrenergic uptake inhibitors and tricyclic antidepressants. CLINICAL IMPLICATIONS: The oral health of people with ADHD may be compromised by inattention and impulsivity that impair home care regimens and can lead to cigarette addiction, which may cause oral cancer and damage the periodontium, and excessive ingestion of caffeinated sugar-laden soft drinks that promote dental caries. To safely care for this patient population, dentists must be familiar with the stimulant and nonstimulant medications used to treat adult ADHD, because these drugs can cause adverse orofacial and systemic reactions and interact adversely with dental therapeutic agents. PMID- 17403738 TI - Calcified carotid artery atheroma and stroke: a systematic review. AB - BACKGROUND: Calcified carotid artery atheroma (CCAA) and its identification on panoramic radiographs have been advocated as a predictor of a cerebrovascular accident (CVA). TYPES OF STUDIES REVIEWED: The authors conducted an electronic search using 11 databases to evaluate the evidence from the literature that links CCAA detection on panoramic radiographs and the precipitation of CVAs among those people. They used the Reporting Recommendations for Tumor Marker Prognostic Studies (REMARK) checklist to perform this systematic review. RESULTS: One study of the 54 studies the authors identified satisfied the REMARK criteria in which CCAA was associated with a negligible increased risk of stroke (95 percent confidence interval, 0 to 0.04 percent) in the population studied. CLINICAL IMPLICATIONS: This systematic review suggests the data supporting the hypothesis that radiographically detectable CCAA is associated with an increased risk of stroke are incomplete and inconclusive. Further research is needed, as clinical guidelines for risk prediction using panoramic radiographs cannot be established on the basis of the current evidence. PMID- 17403739 TI - The relationship between nutrition and systemic and oral well-being in older people. AB - BACKGROUND: Diet plays a key role in disease prevention in older age. The aims of this report were to review the causes and consequences of a poor diet in later life and its interrelationship with oral health. CONCLUSIONS: Energy requirements decrease with age; however, many factors, including poor oral health, loss of appetite and illness, may compromise energy intake. Absorptive capacity may reduce with age; therefore, to prevent deficiencies of micronutrients, a nutrient dense diet is essential. CLINICAL IMPLICATIONS: The first signs of micronutrient deficiencies often are manifest in the oral tissues. Consequently, the dentist has an important role in the early diagnosis of malnutrition. Furthermore, optimizing oral health is important in maximizing older patients' functional capacity to consume a healthful and varied diet. PMID- 17403740 TI - Acute oral ulcerations. PMID- 17403741 TI - Potential inhibition of demineralization in vitro by fluoride-releasing sealants. AB - BACKGROUND: The incorporation of fluoride into sealants has been viewed as a viable way to prevent pit-and-fissure caries by potential inhibition of demineralization through the release of fluoride to enamel. The authors conducted a study to examine the effect of a recently introduced fluoride-releasing sealant (ProSeal, Reliance Orthodontic Products, Itasca, Ill.) on enamel demineralization in an in vitro artificial caries system. METHODS: The authors randomly assigned 45 extracted human third molars to three treatment groups receiving either conventional sealant without fluoride (Group 1), fluoride-releasing sealant (Group 2) or glass ionomer sealant with high fluoride release (Group 3). They placed cavity preparations on the buccal surfaces of the molars and filled them with the assigned material. They placed acid-resistant varnish on the specimens' enamel surfaces to within 1 millimeter of the sealant, leaving a 1-mm rim of sound enamel available for in vitro enamel caries formation. They thermocycled the teeth (500 cycles) in artificial saliva. They subjected the teeth to an in vitro artificial caries challenge for six weeks to produce caries-like lesions in enamel adjacent to the sealant materials. The authors took longitudinal sections from each tooth, immersed them in water and examined them via polarized light microscopy to determine wall lesion frequencies. RESULTS: The mean (+/- standard deviation) lesion depths were 232 +/- 17 micrometers for Group 1, 144 +/- 21 mum for Group 2 and 128 +/- 15 mum for Group 3. The wall lesion frequency was 12 percent for Group 1 and 7 percent for both Groups 2 and 3. There was a significant difference (P < .05) among the fluoride-releasing materials versus the nonfluoride-releasing material. This study indicates that the new fluoride releasing sealant substantially reduces the amount of enamel demineralization adjacent to the material. CONCLUSION: ProSeal provided increased demineralization inhibition compared with a conventional sealant containing no fluoride, but less than that shown by a glass ionomer sealant. CLINICAL IMPLICATIONS: ProSeal's physical properties and cariostatic effects may allow for applications beyond traditional sealant use. PMID- 17403742 TI - A 36-month evaluation of self-etch and etch-and-rinse adhesives in noncarious cervical lesions. AB - BACKGROUND: There are two bonding strategies for composite restorations: the etch and-rinse (ER) approach and the self-etch (SE) approach. Few clinical trials have evaluated the SE approach in Class V restorations for a 36-month period. The authors conducted a study to evaluate whether the SE system can provide retention rates and marginal discoloration similar to that of the ER system. METHODS: Twenty-five patients, each having at least two pairs of equivalent noncarious cervical lesions under occlusion, were enrolled in this study. Two calibrated operators placed 78 restorations, one-half for ER and one-half for SE. Two independent examiners evaluated the restorations at baseline and after six-, 12-, 18- and 36-month periods according to the slightly modified U.S. Public Health Service criteria. Statistical analysis between materials in each period was conducted using a Fisher exact test (alpha = .05), and the performance of the materials at baseline in comparison with each period was evaluated by a McNemar test (alpha = .05). RESULTS: Five SE restorations and one ER restoration were lost after 36 months. After 36 months, 10 SE and five ER restorations were rated Bravo in marginal adaptation (P > .05). Fourteen SE and five ER restorations were rated Bravo in marginal discoloration (P < .05). CONCLUSIONS: Although a significantly increased marginal discoloration was observed with SE, both adhesives showed retention rates in noncarious cervical lesions that were not statistically different after 36 months. CLINICAL IMPLICATIONS: The ER and SE adhesive systems can be used with confidence; however, SE adhesive showed a faster and more progressive enamel marginal degradation. PMID- 17403743 TI - Labial talon cusp: a case study of pre-European-contact American Indians. AB - BACKGROUND: First documented by Mitchell in 1892, the talon cusp is a rare and much-debated morphological variant. The author presents a case study from a pre European-contact American Indian population. In addition, the author addresses debates in the literature regarding the definition, etiology and description of the talon cusp and provides new evidence within an archaeological context. METHODS: The author examined the dentition of 301 skeletons for the presence or absence of talon cusp; five skeletons (2 percent) in the population had the trait. RESULTS: This skeletal sample exhibited variations of the labial talon cusp that can be graded. CONCLUSIONS: The results of this study provide further evidence of the antiquity of labial talon cusp in the assemblage of modern humans and, in particular, in American Indians. PMID- 17403744 TI - Disaster medicine training survey results for dental health care providers in Illinois. AB - BACKGROUND: Ongoing vigilance by governments, public health agencies and health care professionals monitoring potential epidemic and pandemic outbreaks, terrorist threats and ever-present natural disasters requires the continuous evolution of comprehensive disaster response plans and teams, which include the integration of oral health care professionals. METHODS: The authors conducted a study in which oral health care professionals assessed their training in the American Medical Association's (AMA's) National Disaster Life Support (NDLS) courses. At the conclusion of each instructional session, the authors asked participants to complete an anonymous course evaluation form to report their impressions of the training activity. The authors included in the analysis those evaluations associated with sessions attended almost exclusively by dentists and hygienists. RESULTS: The authors derived descriptive statistics from the selected course evaluations. Overall, oral health care professionals believed that the Core Disaster Life Support (CDLS) and Basic Disaster Life Support (BDLS) courses were of great educational value, rating course impact at 9.50 and 9.29, respectively, on a scale from 1 to 10. CONCLUSIONS: Statistical evaluation instruments reveal satisfaction with the all-hazards awareness training received through the AMA's NDLS disaster medicine training curriculum. Licensed oral health care professionals in Illinois accepted the utility and merits of, and benefited from, the four-hour CDLS and eight-hour BDLS certification programs. Practice Implications. Dental professionals in Illinois require minimal additional training for dental emergency responder duties. The AMA's NDLS curriculum provides effective preparation for dental professionals. PMID- 17403745 TI - Laboratories want better impressions. PMID- 17403746 TI - The role of branding in dental practice marketing. PMID- 17403747 TI - What are my ethical obligations in managing a previous dentist's failed treatment? PMID- 17403748 TI - For the dental patient. Braces: straighter teeth can improve oral health. PMID- 17403749 TI - Differing perspectives on patient involvement in patient safety. PMID- 17403750 TI - Making use of mortality data to improve quality and safety in general practice: a review of current approaches. AB - OBJECTIVE: To review studies of the use of mortality data in quality and safety improvement in general practice. DESIGN: Narrative review. METHODS: Search of Medline, Embase and CINAHL for articles reporting mortality monitoring or mortality reviews in general practice. The included articles were reported in English and of any study design, excluding case reports and comment pieces. Studies of palliative care and bereavement, and of primary care programmes in developing countries, were excluded. RESULTS: 229 articles were identified in the searches, 65 were identified as potentially relevant and 53 were included in the review. The studies addressed the impact of primary care provision on mortality rates, methods of monitoring mortality, and the role of audit and death registers in quality and safety improvement. General practitioners were interested in using mortality data but reported difficulties in obtaining complete information. There were no experimental studies of the impact of the use of mortality data, and little evidence of long-term systematic initiatives to use mortality data in quality and safety improvement in general practice. CONCLUSIONS: Mortality data are not used systematically in general practice although general practitioners appear interested in the potential of this information in improving quality and safety. Improved systems to provide complete data are needed and experimental studies required to determine the effectiveness of use of the data to improve general practice care. PMID- 17403752 TI - Implementation of a medication reconciliation process in an ambulatory internal medicine clinic. AB - OBJECTIVE: To evaluate the causes of medication list inaccuracy, implement intervention to enhance overall accuracy of medication lists and measure the sustainability of the intervention. METHODS: A prospective study of patients seen in an academic, ambulatory primary care internal medicine clinic. Before the intervention, baseline data were analysed, assessing completeness of medication documentation in the electronic medical record. The intervention consisted of standardising the entire visit process from scheduling of the appointment to signing of the final clinical note by the physician. Each healthcare team member was instructed in her role to enhance accuracy of the documented medication list. Immediately after the intervention, a second data collection was undertaken to assess the effectiveness of the intervention on the accuracy of individual medications and medication lists. Finally, a year later, a third data collection was undertaken to assess the sustainability of the intervention. RESULTS: Completeness of individual medications improved from 9.7% to 70.7% (p<0.001). However, completeness of the entire medication lists improved only from 7.7% to 18.5%. The incomplete documentation of medication lists was mostly due to lack of route (85.8%) and frequency (22.3%) for individual medications within a medication list. Also, documentation of over-the-counter and "as needed" medications was often incomplete. The incorrectness in a medication list was mostly due to misreporting of medications by patients or failure of clinicians to update the medication list when changes were made. CONCLUSION: To improve the accuracy of medication lists, active participation of all members of the healthcare team and the patient is needed. PMID- 17403753 TI - Confidential reporting of patient safety events in primary care: results from a multilevel classification of cognitive and system factors. AB - OBJECTIVE: To classify events of actual or potential harm to primary care patients using a multilevel taxonomy of cognitive and system factors. METHODS: Observational study of patient safety events obtained via a confidential but not anonymous reporting system. Reports were followed up with interviews where necessary. Events were analysed for their causes and contributing factors using causal trees and were classified using the taxonomy. Five general medical practices in the West Midlands were selected to represent a range of sizes and types of patient population. All practice staff were invited to report patient safety events. Main outcome measures were frequencies of clinical types of events reported, cognitive types of error, types of detection and contributing factors; and relationship between types of error, practice size, patient consequences and detection. RESULTS: 78 reports were relevant to patient safety and analysable. They included 21 (27%) adverse events and 50 (64%) near misses. 16.7% (13/71) had serious patient consequences, including one death. 75.7% (59/78) had the potential for serious patient harm. Most reports referred to administrative errors (25.6%, 20/78). 60% (47/78) of the reports contained sufficient information to characterise cognition: "situation assessment and response selection" was involved in 45% (21/47) of these reports and was often linked to serious potential consequences. The most frequent contributing factor was work organisation, identified in 71 events. This included excessive task demands (47%, 37/71) and fragmentation (28%, 22/71). CONCLUSIONS: Even though most reported events were near misses, events with serious patient consequences were also reported. Failures in situation assessment and response selection, a cognitive activity that occurs in both clinical and administrative tasks, was related to serious potential harm. PMID- 17403754 TI - Lag time in an incident reporting system at a university hospital in Japan. AB - BACKGROUND: Delays and underreporting limit the success of hospital incident reporting systems, but little is known about the causes or implications of delayed reporting. SETTING AND METHODS: The authors examined 6880 incident reports filed by physicians and nurses for three years at a national university hospital in Japan and evaluated the lag time between each incident and the submission of a report. RESULTS: Although physicians and nurses reported nearly equal numbers of events resulting in major injury (32 v 31), physicians reported far fewer minor incidents (430 v 6387) and far fewer incidents overall (462 v 6418). In univariate analyses, lag time was significantly longer for physicians than nurses (3.79 v 2.20 days; p<0.001). In multivariate analysis, physicians had adjusted reporting lag time 75% longer than nurses (p<0.001) and lag time for major injuries was 18% shorter than for minor injuries (p = 0.011). Adjusted lag time in 2002 and 2004 were 34% longer than in 2003 (p<0.001). CONCLUSIONS: Physicians report fewer incidents than nurses and take longer to report them. Quantitative evaluation of lag time may facilitate improvements in incident reporting systems by distinguishing institutional obstacles to physician reporting from physicians' lesser willingness to report. PMID- 17403755 TI - Antibiotic prescribing in primary care: first choice and restrictive prescribing are two different traits. AB - OBJECTIVE: To investigate the quality of antibiotic prescribing in primary care using quality indicators and the relatedness of these indicators. To determine the influence of general practice and practice population characteristics on the indicator scores. METHODS: Data on performance were collected during the Second National Survey of General Practice over 1 year between May 2000 and April 2002 in The Netherlands. The study was carried out in 104 computerised general practices, comprising 195 general practitioners and about 400,000 patients. From a preliminary set of quality indicators on antibiotic prescribing (n = 15), eight were selected covering various medical conditions. Indicator scores were derived. A factor analysis was performed to examine the relatedness of these indicators. Composite scores were calculated for the indicators loading on the same factor. The influence of general practice and practice population characteristics on the quality of antibiotic prescribing was investigated. RESULTS: Considerable variation was found between indicator scores (32.8-94.2%) and between practices. The factor analysis discovered two interpretable factors-namely, "first choice prescribing" and "restrictive prescribing". The composite scores were 64% and 68%, respectively. No significant correlation was found between the two composite scores. Practice and population characteristics explained only a small proportion of the variance between practices. CONCLUSIONS: Although different quality indicators on antibiotic prescribing are grouped together over several medical conditions, there is large variation between those indicators. General practices performing well on first choice prescribing do not automatically perform well on restrictive prescribing. There is room for improvement on both aspects of prescribing. The variation between practices is clearly present and should be further investigated. PMID- 17403756 TI - An integrated care pathway for menorrhagia across the primary-secondary interface: patients' experience, clinical outcomes, and service utilisation. AB - BACKGROUND: "Referral" characterises a significant area of interaction between primary and secondary care. Despite advantages, it can be inflexible, and may lead to duplication. OBJECTIVE: To examine the outcomes of an integrated model that lends weight to general practitioner (GP)-led evidence based care. DESIGN: A prospective, non-random comparison of two services: women attending the new (Bridges) pathway compared with those attending a consultant-led one-stop menstrual clinic (OSMC). Patients' views were examined using patient career diaries, health and clinical outcomes, and resource utilisation. Follow-up was for 8 months. SETTING: A large teaching hospital and general practices within one primary care trust (PCT). RESULTS: Between March 2002 and June 2004, 99 women in the Bridges pathway were compared with 94 women referred to the OSMC by GPs from non-participating PCTs. The patient career diary demonstrated a significant improvement in the Bridges group for patient information, fitting in at the point of arrangements made for the patient to attend hospital (ease of access) (p<0.001), choice of doctor (p = 0.020), waiting time for an appointment (p<0.001), and less "limbo" (patient experience of non-coordination between primary and secondary care) (p<0.001). At 8 months there were no significant differences between the two groups in surgical and medical treatment rates or in the use of GP clinic appointments. Significantly fewer (traditional) hospital outpatient appointments were made in the Bridges group than in the OSMC group (p<0.001). CONCLUSION: A general practice-led model of integrated care can significantly reduce outpatient attendance while improving patient experience, and maintaining the quality of care. PMID- 17403758 TI - Medication errors in paediatric care: a systematic review of epidemiology and an evaluation of evidence supporting reduction strategy recommendations. AB - BACKGROUND: Although children are at the greatest risk for medication errors, little is known about the overall epidemiology of these errors, where the gaps are in our knowledge, and to what extent national medication error reduction strategies focus on children. OBJECTIVE: To synthesise peer reviewed knowledge on children's medication errors and on recommendations to improve paediatric medication safety by a systematic literature review. DATA SOURCES: PubMed, Embase and Cinahl from 1 January 2000 to 30 April 2005, and 11 national entities that have disseminated recommendations to improve medication safety. STUDY SELECTION: Inclusion criteria were peer reviewed original data in English language. Studies that did not separately report paediatric data were excluded. DATA EXTRACTION: Two reviewers screened articles for eligibility and for data extraction, and screened all national medication error reduction strategies for relevance to children. DATA SYNTHESIS: From 358 articles identified, 31 were included for data extraction. The definition of medication error was non-uniform across the studies. Dispensing and administering errors were the most poorly and non uniformly evaluated. Overall, the distributional epidemiological estimates of the relative percentages of paediatric error types were: prescribing 3-37%, dispensing 5-58%, administering 72-75%, and documentation 17-21%. 26 unique recommendations for strategies to reduce medication errors were identified; none were based on paediatric evidence. CONCLUSIONS: Medication errors occur across the entire spectrum of prescribing, dispensing, and administering, are common, and have a myriad of non-evidence based potential reduction strategies. Further research in this area needs a firmer standardisation for items such as dose ranges and definitions of medication errors, broader scope beyond inpatient prescribing errors, and prioritisation of implementation of medication error reduction strategies. PMID- 17403759 TI - Ambulatory care adverse events and preventable adverse events leading to a hospital admission. AB - BACKGROUND: Most healthcare in the US is delivered in the ambulatory care setting, but the epidemiology of errors and adverse events in ambulatory care is understudied. METHODS: Using the population-based data from the Colorado and Utah Medical Practices Study, we identified adverse events that occurred in an ambulatory care setting and led to hospital admission. Proportions with 95% CIs are reported. RESULTS: We reviewed 14,700-hospital discharge records and found 587 adverse events of which 70 were ambulatory care adverse events (AAEs) and 31 were ambulatory care preventable adverse events (APAEs). When weighted to the general population, there were 2608 AAEs and 1296 (44.3%) APAEs in Colorado and Utah, USA, in 1992. APAEs occurred most commonly in physicians' offices (43.1%, range 46.8-27.8), the emergency department (32.3%, 46.1-18.5) and at home (13.1%, 23.1-3.1). APAEs in day surgery were less common (7.1%, 13.6-0.6) but caused the greatest harm to patients. The types of APAEs were broadly distributed among missed or delayed diagnoses (36%, 50.2-21.8), surgery (24.1%, 36.7-11.5), non surgical procedures (14.6%, 25.0-4.2), medication (13.1%, 23.1-3.1) and therapeutic events (12.3%, 22.0-2.6). Overall, 10% of the APAEs resulted in serious permanent injury or death. The proportion of APAEs that resulted in death was 31.8% for general internal medicine, 22.5% for family practice and 16.7% for emergency medicine. CONCLUSION: An estimated 75,000 hospitalisations per year are due to preventable adverse events that occur in outpatient settings in the US, resulting in 4839 serious permanent injuries and 2587 deaths. PMID- 17403760 TI - Using trigger phrases to detect adverse drug reactions in ambulatory care notes. AB - BACKGROUND: As medical care moves towards an outpatient focus, monitoring systems for ambulatory patients are increasingly important. Because adverse outcomes due to medications are an important problem in outpatients, the authors developed an automated monitoring system for detecting adverse drug reactions (ADRs) in ambulatory patients. METHODS: The authors obtained a set of approximately 110,000 ambulatory care notes from the medicine clinic at Bellevue Hospital Centre for 2003-4, and manually analysed a representative sample of 1250 notes to obtain a gold standard. To detect ADRs in the text of electronic ambulatory notes, the authors used a "trigger phrases" methodology, based on a simple grammar populated with a limited set of keywords. RESULTS: Under current functionality, this system detected 38 of 54 cases in the authors' gold standard set, of which 17 were true positives, for a sensitivity of 31%, a specificity of 98%, and a positive predictive value of 45%. Their proxy measure correlated with 70% of the ADRs in the gold standard. These values are comparable or superior to other systems described in the literature. CONCLUSIONS: These results show that an automated system can detect ADRs with moderate sensitivity and high specificity, and has the potential to serve as the basis for a larger scale reporting system. PMID- 17403761 TI - Quantifying distraction and interruption in urological surgery. AB - BACKGROUND: To enhance safety in surgery, it is necessary to develop a variety of tools for measuring and evaluating the system of work. One important consideration for safety in any high-risk work is the frequency and effect of distraction and interruption. AIM: To quantify distraction and interruption to the sterile surgical team in urology. METHODS: Observation of the behaviour of the surgical team and their task activity determined distraction and interruption recorded. Using an ordinal scale, an observer rated each salient distraction or interruption observed in relation to the team's involvement. RESULTS: The frequency of events and their attached ratings were high, deriving from varying degrees of equipment, procedure and environment problems, telephones, bleepers and conversations. DISCUSSION: With further refinement and testing, this method may be useful for distinguishing ordinal levels of work interference in surgery and helpful in raising awareness of its origin for postoperative surgical team debriefing. PMID- 17403763 TI - Should patients have a role in patient safety? A safety engineering view. PMID- 17403764 TI - Barriers to optimal antibiotic use for community-acquired pneumonia at hospitals: a qualitative study. AB - BACKGROUND: Physician adherence to key recommendations of guidelines for community-acquired pneumonia (CAP) is often not optimal. A better understanding of factors influencing optimal performance is needed to plan effective change. METHODS: The authors used semistructured interviews with care providers in three Dutch medium-sized hospitals to qualitatively study and understand barriers to appropriate antibiotic use in patients with CAP. They discussed recommendations about the prescription of empirical antibiotic therapy that adheres to the guidelines, timely administration of antibiotics, adjusting antibiotic dosage to accommodate decreased renal function, switching and streamlining therapy, and blood and sputum culturing. The authors then classified the barriers each recommendation faced into categories using a conceptual framework (Cabana). RESULTS: Eighteen interviews were performed with residents and specialists in pulmonology and internal medicine, with medical microbiologists and a clinical pharmacist. Two additional multidisciplinary small group interviews which included nurses were performed. Each guideline recommendation elicited a different type of barrier. Regarding the choice of guideline-adherent empirical therapy, treating physicians said that they worried about patient outcome when prescribing narrow-spectrum antibiotic therapy. Regarding the timeliness of antibiotic administration, barriers such as conflicting guidelines and organisational factors (for example, delayed laboratory results, antibiotics not directly available, lack of time) were reported. Not streamlining therapy after culture results became available was thought to be due to the physicians' attitude of "never change a winning team". CONCLUSIONS: Efforts to improve the use of antibiotics for patients with CAP should consider the range of barriers that care providers face. Each recommendation meets its own barriers. Interventions to improve adherence should be tailored to these factors. PMID- 17403765 TI - Development and testing of an assessment instrument for the formative peer review of significant event analyses. AB - AIM: To establish the content validity and specific aspects of reliability for an assessment instrument designed to provide formative feedback to general practitioners (GPs) on the quality of their written analysis of a significant event. METHODS: Content validity was quantified by application of a content validity index. Reliability testing involved a nested design, with 5 cells, each containing 4 assessors, rating 20 unique significant event analysis (SEA) reports (10 each from experienced GPs and GPs in training) using the assessment instrument. The variance attributable to each identified variable in the study was established by analysis of variance. Generalisability theory was then used to investigate the instrument's ability to discriminate among SEA reports. RESULTS: Content validity was demonstrated with at least 8 of 10 experts endorsing all 10 items of the assessment instrument. The overall G coefficient for the instrument was moderate to good (G>0.70), indicating that the instrument can provide consistent information on the standard achieved by the SEA report. There was moderate inter-rater reliability (G>0.60) when four raters were used to judge the quality of the SEA. CONCLUSIONS: This study provides the first steps towards validating an instrument that can provide educational feedback to GPs on their analysis of significant events. The key area identified to improve instrument reliability is variation among peer assessors in their assessment of SEA reports. Further validity and reliability testing should be carried out to provide GPs, their appraisers and contractual bodies with a validated feedback instrument on this aspect of the general practice quality agenda. PMID- 17403766 TI - The economic burden of complications during percutaneous coronary intervention. AB - BACKGROUND: Technological advances have enabled percutaneous coronary intervention (PCI) to be applied with expanding indications. However, escalating costs are of concern. This study assessed the incremental medical costs of major in-hospital procedural complications incurred by patients undergoing PCI. METHODS: We considered all patients undergoing elective, urgent, or emergent PCI at Mayo Clinic Rochester between 3/1/1998-3/31/2003 in analyses. Clinical, angiographic, and outcome data were derived from the Mayo Clinic PCI Registry. In hospital PCI complications included major adverse cardiac and cerebrovascular events (MACCE) and bleeding of clinical significance. Administrative data were used to estimate total costs in standardised, year 2004, constant-US dollars. We used generalised linear modeling to estimate costs associated with complications adjusting for baseline and procedural characteristics. RESULTS: 1071 (13.2%) of patients experienced complications during hospitalisation. Patients experiencing complications were older, more likely to present with emergent PCI, recent or prior myocardial infarction, multi-vessel disease, and comorbid conditions than patients who did not experience these events. Unadjusted total costs were, on average, $27,865+/-$39,424 for complicated patient episodes compared to $12,279+/ $6796 for episodes that were complication free (p<0.0001). Adjusted mean costs were $6984 higher for complicated PCIs compared with uncomplicated PCI episodes (95% CI of cost difference: $5801, $8168). Incremental costs associated with isolated bleeding events, MACCE, or for both bleeding and MACCE events were $5883, $5086, and $15,437, respectively (p<0.0001). CONCLUSIONS: This high-volume study highlights the significant economic burden associated with procedural complications. Resources and systems approaches to minimising clinical and economic complications in PCI are warranted. PMID- 17403768 TI - Identifying patients in hospital: are more adverse events waiting to happen? PMID- 17403769 TI - Nonspecific PCR amplification of the 16S rRNA gene segment in different bacteria by use of primers specific for Campylobacter, Arcobacter, and Helicobacter spp. PMID- 17403770 TI - Emergence of non-ceftriaxone-susceptible Neisseria meningitidis in India. PMID- 17403771 TI - Skewed expression and up-regulation of the IL-12 and IL-18 receptors in resting and activated CD4 T cells from HIV-1-infected patients. AB - IL-12 and IL-18 synergistically induce the production of IFN-gamma by resting and activated T cells. To evaluate whether this induction was affected in HIV-1 infected patients, PBMC or isolated CD4 T cells were cultured with IL-12 plus IL 18, anti-CD3 plus anti-CD28, or PHA for 72 h. Cell samples were labeled daily to assess the levels of IL-12 receptor beta1 (IL-12Rbeta1), IL-12Rbeta2, and IL 18Ralpha. Culture supernatants were analyzed for the presence of Th1- and Th2 related cytokines by ELISA or cytometric bead array and analyzed by flow cytometry. A twofold increase in the percentage of CD4-resting T cells expressing IL-12Rbeta1 and IL-18Ralpha from HIV-1-infected patients was observed when compared with cells from HIV-1-negative donors. Higher IL-12Rbeta1 and IL 18Ralpha expression correlated (r=0.87; P<0.007) to increased production of IFN gamma by isolated CD4 T cells in the presence of IL-12 and IL-18. Moreover, exogenous IL-12 and IL-18 induced the up-regulation of IL-12Rbeta2 to twice higher in CD4 T cells from HIV-1-positive individuals compared with controls. Conversely, upon activation with anti-CD3 and anti-CD28 antibodies, only 25% of the CD4+ T cells from HIV-1 patients showed an increase in the IL-12beta2 when compared with 50% in healthy controls. Furthermore, the percentage of IL-12Rbeta1 positive cells correlated inversely with the CD4 nadir of patients, suggesting that deregulation of the IL-12 and IL-18 pathways may play a role in the immunopathogenesis of HIV-1 infection. PMID- 17403772 TI - MDP-induced interleukin-1beta processing requires Nod2 and CIAS1/NALP3. AB - Nucleotide-binding oligomerization domain (Nod)2 is a sensor of muramyl dipeptides (MDP) derived from bacterial peptidoglycan. Nod2 also plays a role in some autoinflammatory diseases. Cold-induced autoinflammatory syndrome 1 (CIAS1)/NACHT domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NALP3) has been suggested to be sufficient for MDP-dependent release of mature IL-1beta, but the role of Nod2 in this process is unclear. Using mice bearing selective gene deletions, we provide in vitro and in vivo data showing that MDP induced IL-1beta release requires Nod2 and CIAS1/NALP3 as well as receptor interacting protein-2 (Rip2), apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC), and caspase-1. In contrast, MDP-dependent IL-6 production only requires Nod2 and Rip2. Together, our data provide a new understanding of this important pathway of IL-1beta production and allow for further studies of the role of these proteins within the broader context of inflammatory disease. PMID- 17403773 TI - NBK/BIK antagonizes MCL-1 and BCL-XL and activates BAK-mediated apoptosis in response to protein synthesis inhibition. AB - Ribonucleases, antibiotics, bacterial toxins, and viruses inhibit protein synthesis, which results in apoptosis in mammalian cells. How the BCL-2 family of proteins regulates apoptosis in response to the shutoff of protein synthesis is not known. Here we demonstrate that an Escherichia coli toxin, MazF, inhibited protein synthesis by cleavage of cellular mRNA and induced apoptosis in mammalian cells. MazF-induced apoptosis required proapoptotic BAK and its upstream regulator, the proapoptotic BH3-only protein NBK/BIK, but not BIM, PUMA, or NOXA. Interestingly, in response to MazF induction, NBK/BIK activated BAK by displacing it from anti-apoptotic proteins MCL-1 and BCL-X(L) that sequester BAK. Furthermore, NBK/BIK- or BAK-deficient cells were resistant to cell death induced by pharmacologic inhibition of translation and by virus-mediated shutoff of protein synthesis. Thus, the BH3-only protein NBK/BIK is the apical regulator of a BAK-dependent apoptotic pathway in response to shutoff of protein synthesis that functions to displace BAK from sequestration by MCL1 and BCL-X(L). Although NBK/BIK is dispensable for development, it is the BH3-only protein targeted for inactivation by viruses, suggesting that it plays a role in pathogen/toxin response through apoptosis activation. PMID- 17403774 TI - Discovery of tMAC: a Drosophila testis-specific meiotic arrest complex paralogous to Myb-Muv B. AB - The Drosophila Myb-Muv B (MMB)/dREAM complex regulates gene expression and DNA replication site-specifically, but its activities in vivo have not been thoroughly explored. In ovarian amplification-stage follicle cell nuclei, the largest subunit, Mip130, is a negative regulator of replication, whereas another subunit, Myb, is a positive regulator. Here, we identified a mutation in mip40 and generated a mutation in mip120, two additional MMB subunits. Both mutants were viable, but mip120 mutants had many complex phenotypes including shortened longevity and severe eye defects. mip40 mutant females had severely reduced fertility, whereas mip120 mutant females were sterile, substantiating ovarian regulatory role(s) for MMB. Myb accumulation and binding to polytene chromosomes was dependent on the core factors of the MMB complex. In contrast to the documented mip130 mutant phenotypes, both mip40 and mip120 mutant males were sterile. We purified Mip40-containing complexes from testis nuclear extracts and identified tMAC, a new testis-specific meiotic arrest complex that contained Mip40, Caf1/p55, the Mip130 family member, Always early (Aly), and a Mip120 family member, Tombola (Tomb). Together, these data demonstrate that MMB serves diverse roles in different developmental pathways, and members of MMB can be found in alternative, noninteracting complexes in different cell types. PMID- 17403775 TI - H2B ubiquitylation in transcriptional control: a FACT-finding mission. PMID- 17403776 TI - The microRNA miR-124 antagonizes the anti-neural REST/SCP1 pathway during embryonic CNS development. AB - Neuronal gene expression is tightly regulated in developing CNS. Here, we demonstrate the anti-neural function of phosphatase SCP1 (small C-terminal domain phosphatase 1) during development. We further show that the neuron-enriched microRNA miR-124 directly targets SCP1-3' untranslated region (UTR) to suppress SCP1 expression. In developing spinal cord, expression of miR-124 and SCP1 is complementary, and miR-124 antagonism phenocopies SCP1 overexpression and vice versa. In P19 cells, miR-124 suppresses SCP1 expression and induces neurogenesis, and SCP1 counteracts this proneural activity of miR-124. Our results suggest that, during CNS development, timely down-regulation of SCP1 is critical for inducing neurogenesis, and miR-124 contributes to this process at least in part by down-regulating SCP1 expression. PMID- 17403777 TI - Two small regulatory RNAs establish opposing fates of a developmental axis. AB - Small RNAs are important regulators of gene expression. In maize, adaxial/abaxial (dorsoventral) leaf polarity is established by an abaxial gradient of microRNA166 (miR166), which spatially restricts the expression domain of class III homeodomain leucine zipper (HD-ZIPIII) transcription factors that specify adaxial/upper fate. Here, we show that leafbladeless1 encodes a key component in the trans-acting small interfering RNA (ta-siRNA) biogenesis pathway that acts on the adaxial side of developing leaves and demarcates the domains of hd-zipIII and miR166 accumulation. Our findings indicate that tasiR-ARF, a ta-siRNA, and miR166 establish opposing domains along the adaxial-abaxial axis, thus revealing a novel mechanism of pattern formation. PMID- 17403778 TI - Expansion of adult beta-cell mass in response to increased metabolic demand is dependent on HNF-4alpha. AB - The failure to expand functional pancreatic beta-cell mass in response to increased metabolic demand is a hallmark of type 2 diabetes. Lineage tracing studies indicate that replication of existing beta-cells is the principle mechanism for beta-cell expansion in adult mice. Here we demonstrate that the proliferative response of beta-cells is dependent on the orphan nuclear receptor hepatocyte nuclear factor-4alpha (HNF-4alpha), the gene that is mutated in Maturity-Onset Diabetes of the Young 1 (MODY1). Computational analysis of microarray expression profiles from isolated islets of mice lacking HNF-4alpha in pancreatic beta-cells reveals that HNF-4alpha regulates selected genes in the beta-cell, many of which are involved in proliferation. Using a physiological model of beta-cell expansion, we show that HNF-4alpha is required for beta-cell replication and the activation of the Ras/ERK signaling cascade in islets. This phenotype correlates with the down-regulation of suppression of tumorigenicity 5 (ST5) in HNF-4alpha mutants, which we identify as a novel regulator of ERK phosphorylation in beta-cells and a direct transcriptional target of HNF-4alpha in vivo. Together, these results indicate that HNF-4alpha is essential for the physiological expansion of adult beta-cell mass in response to increased metabolic demand. PMID- 17403779 TI - PGC-1alpha regulates the neuromuscular junction program and ameliorates Duchenne muscular dystrophy. AB - The coactivator PGC-1alpha mediates key responses of skeletal muscle to motor nerve activity. We show here that neuregulin-stimulated phosphorylation of PGC 1alpha and GA-binding protein (GABP) allows recruitment of PGC-1alpha to the GABP complex and enhances transcription of a broad neuromuscular junction gene program. Since a subset of genes controlled by PGC-1alpha and GABP is dysregulated in Duchenne muscular dystrophy (DMD), we examined the effects of transgenic PGC-1alpha in muscle of mdx mice. These animals show improvement in parameters characteristic of DMD, including muscle histology, running performance, and plasma creatine kinase levels. Thus, control of PGC-1alpha levels in skeletal muscle could represent a novel avenue to prevent or treat DMD. PMID- 17403780 TI - Rhythmic control of AANAT translation by hnRNP Q in circadian melatonin production. AB - The circadian rhythm of pineal melatonin requires the nocturnal increment of serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase [AANAT]) protein. To date, only limited information is available in the critical issue of how AANAT protein expression is up-regulated exclusively at night regardless of its species-specific mRNA profiles. Here we show that the circadian timing of AANAT protein expression is regulated by rhythmic translation of AANAT mRNA. This rhythmic control is mediated by both a highly conserved IRES (internal ribosome entry site) element within the AANAT 5' untranslated region and its partner hnRNP Q (heterogeneous nuclear ribonucleoprotein Q) with a peak in the middle of the night. Consistent with the enhancing role of hnRNP Q in AANAT IRES activities, knockdown of the hnRNP Q level elicited a dramatic decrease of peak amplitude in the AANAT protein profile parallel to reduced melatonin production in pinealocytes. This translational regulation of AANAT mRNA provides a novel aspect for achieving the circadian rhythmicity of vertebrate melatonin. PMID- 17403781 TI - Rearrangement of competing U2 RNA helices within the spliceosome promotes multiple steps in splicing. AB - Nuclear pre-messenger RNA (pre-mRNA) splicing requires multiple spliceosomal small nuclear RNA (snRNA) and pre-mRNA rearrangements. Here we reveal a new snRNA conformational switch in which successive roles for two competing U2 helices, stem IIa and stem IIc, promote distinct splicing steps. When stem IIa is stabilized by loss of stem IIc, rapid ATP-independent and Cus2p-insensitive prespliceosome formation occurs. In contrast, hyperstabilized stem IIc improves the first splicing step on aberrant branchpoint pre-mRNAs and rescues temperature sensitive U6-U57C, a U6 mutation that also suppresses first-step splicing defects of branchpoint mutations. A second, later role for stem IIa is revealed by its suppression of a cold-sensitive allele of the second-step splicing factor PRP16. Our data expose a spliceosomal progression cycle of U2 stem IIa formation, disruption by stem IIc, and then reformation of stem IIa before the second catalytic step. We propose that the competing stem IIa and stem IIc helices are key spliceosomal RNA elements that optimize juxtaposition of the proper reactive sites during splicing. PMID- 17403782 TI - U2 toggles iteratively between the stem IIa and stem IIc conformations to promote pre-mRNA splicing. AB - To ligate exons in pre-messenger RNA (pre-mRNA) splicing, the spliceosome must reposition the substrate after cleaving the 5' splice site. Because spliceosomal small nuclear RNAs (snRNAs) bind the substrate, snRNA structures may rearrange to reposition the substrate. However, such rearrangements have remained undefined. Although U2 stem IIc inhibits binding of U2 snRNP to pre-mRNA during assembly, we found that weakening U2 stem IIc suppressed a mutation in prp16, a DExD/H box ATPase that promotes splicing after 5' splice site cleavage. The prp16 mutation was also suppressed by mutations flanking stem IIc, suggesting that Prp16p facilitates a switch from stem IIc to the mutually exclusive U2 stem IIa, which activates binding of U2 to pre-mRNA during assembly. Providing evidence that stem IIa switches back to stem IIc before exon ligation, disrupting stem IIa suppressed 3' splice site mutations, and disrupting stem IIc impaired exon ligation. Disrupting stem IIc also exacerbated the 5' splice site cleavage defects of certain substrate mutations, suggesting a parallel role for stem IIc at both catalytic stages. We propose that U2, much like the ribosome, toggles between two conformations--a closed stem IIc conformation that promotes catalysis and an open stem IIa conformation that promotes substrate binding and release. PMID- 17403783 TI - HLA-B-associated transcript 3 (Bat3)/Scythe is essential for p300-mediated acetylation of p53. AB - In response to DNA damage, p53 undergoes post-translational modifications (including acetylation) that are critical for its transcriptional activity. However, the mechanism by which p53 acetylation is regulated is still unclear. Here, we describe an essential role for HLA-B-associated transcript 3 (Bat3)/Scythe in controlling the acetylation of p53 required for DNA damage responses. Depletion of Bat3 from human and mouse cells markedly impairs p53 mediated transactivation of its target genes Puma and p21. Although DNA damage induced phosphorylation, stabilization, and nuclear accumulation of p53 are not significantly affected by Bat3 depletion, p53 acetylation is almost completely abolished. Bat3 forms a complex with p300, and an increased amount of Bat3 enhances the recruitment of p53 to p300 and facilitates subsequent p53 acetylation. In contrast, Bat3-depleted cells show reduced p53-p300 complex formation and decreased p53 acetylation. Furthermore, consistent with our in vitro findings, thymocytes from Bat3-deficient mice exhibit reduced induction of puma and p21, and are resistant to DNA damage-induced apoptosis in vivo. Our data indicate that Bat3 is a novel and essential regulator of p53-mediated responses to genotoxic stress, and that Bat3 controls DNA damage-induced acetylation of p53. PMID- 17403784 TI - Decline in ribosomal fidelity contributes to the accumulation and stabilization of the master stress response regulator sigmaS upon carbon starvation. AB - The sigma(S) subunit of RNA polymerase is a master regulator of Escherichia coli that retards cellular senescence and bestows cells with general stress protective functions during growth arrest. We show that mutations and drugs triggering translational errors elevate sigma(S) levels and stability. Furthermore, mutations enhancing translational fidelity attenuate induction of the rpoS regulon and prevent stabilization of sigma(S) upon carbon starvation. Destabilization of sigma(S) by increased proofreading requires the presence of the sigma(S) recognition factor SprE (RssB) and the ClpXP protease. The data further suggest that sigma(S) becomes stabilized upon starvation as a result of ClpP sequestration and this sequestration is enhanced by oxidative modifications of aberrant proteins produced by erroneous translation. ClpP overproduction counteracted starvation-induced stabilization of sigma(S), whereas overproduction of a ClpXP substrate (ssrA-tagged GFP) stabilized sigma(S) in exponentially growing cells. We present a model for the sequence of events leading to the accumulation and activation of sigma(S) upon carbon starvation, which are linked to alterations in both ribosomal fidelity and efficiency. PMID- 17403786 TI - Social determinants of ever initiating smoking differ from those of quitting: a cross-sectional study in Estonia. AB - BACKGROUND: To study the association of socioeconomic and demographic indicators with ever initiating regular smoking and quitting smoking among ever regular smokers in Estonia in order to identify target groups for equity-oriented tobacco control policies. METHODS: Data for 4277 individuals in the 25-64 age group come from three cross-sectional studies conducted in 2000, 2002 and 2004. Age standardized prevalence rates and odds ratios with 95% confidence intervals were calculated. RESULTS: High rates of ever initiation were observed for lower educated men and women after controlling for other socioeconomic indicators. This association was not observed for women above age 50. Independent, although weaker associations were found among men who were unemployed or with a low occupational status. Low cessation rates were observed among men who were unemployed, who had a lower occupational position or who had a low income. These associations remained after controlling for other socioeconomic variables. The effect of income became stronger in the older age groups among men. Rates of ever initiation and cessation also varied strongly in relationship to some demographic variables. The highest initiation rates were found among divorced women and among women living in the capital city. The lowest cessation rates were found among divorced women, and among Russian men. CONCLUSIONS: While educational level was the strongest predictor of ever initiating regular smoking, smoking cessation was related more directly to aspects of social disadvantage originating in adult life. To be effective, tobacco control interventions should not only target lower educated, but also those in material disadvantage. PMID- 17403787 TI - Increasing life expectancy in Germany: quantitative contributions from changes in age- and disease-specific mortality. AB - BACKGROUND: Between 1962 and 2002 the average life expectancy in Germany has increased from 67.1 years to 75.6 years in men and from 72.7 years to 81.3 in women. METHODS: The cumulative and annual contributions of different age- and disease-groups on life expectancy were calculated using Pollard's actuarial method of decomposing mortality rates. Mortality data were provided by the German Statistical Office. RESULTS: Considering the cumulative contribution over the period of 40 years, the largest contributions came from persons with at least 65 years of age (2.9 years in men and 4.0 years in women). Reductions in cardiovascular disease mortality had the greatest cumulative impact on life expectancy (2.7 years in men and 3.0 years in women). The contribution from reduced cancer mortality on life expectancy was substantially lower (0.6 and 0.9 years, respectively). The annual contributions of several disease-groups varied considerably over time. The positive contribution from cardiovascular diseases started only after 1970, and in men it became solid only after 1980. Regarding malignant neoplasms, the largest cumulative contribution came from stomach cancer (0.4 in both sexes). The annual analyses showed increasing contributions from reduced cancer mortality after 1990. These were strongly influenced by lung, stomach, prostate and colorectal cancer in men, and by breast, colorectal and stomach cancer in women. CONCLUSIONS: While life expectancy has increased by about 2.2 years per decade the observed variations in the age- and disease specific contributions over time have implications for future health care planning and prevention strategies. PMID- 17403788 TI - Estimating mortality and causes of death in Turkey: methods, results and policy implications. AB - BACKGROUND: Cause-specific mortality statistics are primary evidence for health policy formulation, programme evaluation, and epidemiological research. In Turkey, a partially functioning vital registration system in urban areas yields fragmentary evidence on levels and causes of mortality. This article discusses the application of innovative methods to develop national mortality estimates in Turkey, and their implications for national health development policies. METHODS: Child mortality levels from the Demography and Health Survey (DHS) were applied to model life tables to estimate age-specific death rates. Reported causes of death from urban areas were adjusted using re-distribution algorithms from the Global Burden of Disease (GBD) Study. Rural cause structure was estimated from epidemiological models. Local epidemiological data was used to adjust model-based estimates. RESULTS: Life expectancy at birth in 2000 was estimated to be 67.7 years (males) and 71.9 years (females), about 8-10 years lower than in Western Europe. Leading causes of death include major vascular diseases (ischaemic heart disease, stroke) causing 35-38% of deaths, chronic obstructive lung disease and lung cancer in men, but also perinatal causes, lower respiratory infections and diarrhoeal diseases. Injuries cause about 6-8% of deaths, although this may be an underestimate. CONCLUSIONS: Mortality estimates are uncertain in Turkey, given the poor quality of death registration systems. Application of burden of disease methods suggests that there has been progress along the epidemiological transition. Key health development strategies for Turkey include improved access to communicable disease control technologies, and urgent attention to the development of a reliable, nationally representative health information system. PMID- 17403789 TI - Timing of tourniquet release in knee arthroplasty. Meta-analysis of randomized, controlled trials. AB - BACKGROUND: Some surgeons release the tourniquet before closing the wound to secure hemostasis during knee arthroplasty. We examined whether early tourniquet release could reduce the perioperative blood loss and whether not releasing the tourniquet until after wound closure would increase the risk of early postoperative complications. METHODS: We searched electronic databases and reference lists of relevant articles, retrieved all of the published randomized controlled trials designed to address these issues, and performed a meta analysis. RESULTS: Eleven studies involving a total of 872 patients and 893 primary knee arthroplasties were analyzed systematically. The studies showed considerable clinical and methodological diversity. Early release of the tourniquet increased the total measured blood loss (weighted mean difference = 228.7 mL; 95% confidence interval = 168.3 to 289.1; p < 0.00001). Early release also increased blood loss as calculated on the basis of the maximum decrease in hemoglobin concentration (weighted mean difference = 320.7 mL; 95% confidence interval = 204.3 to 437.1; p < 0.00001). The rate of reoperations due to postoperative complications was 3.1% (nine of 290) in the group with late tourniquet release compared with 0.3% (one of 290) in the group with early tourniquet release; the risk difference was 3% (95% confidence interval, 0.1% to 5%), which was significant (p = 0.04). CONCLUSIONS: Early tourniquet release for hemostasis increases the blood loss associated with primary knee arthroplasty. However, tourniquet release after wound closure can increase the risk of early postoperative complications requiring another operation. Well-conducted large studies are needed to further explore the risk of early postoperative complications associated with late tourniquet release in knee arthroplasty. LEVEL OF EVIDENCE: Therapeutic Level II. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403790 TI - Lateral entry compared with medial and lateral entry pin fixation for completely displaced supracondylar humeral fractures in children. A randomized clinical trial. AB - BACKGROUND: Closed reduction and percutaneous pin fixation is the treatment of choice for completely displaced (type-III) extension supracondylar fractures of the humerus in children, although controversy persists regarding the optimal pin fixation technique. The purpose of this study was to compare the efficacy of lateral entry pin fixation with that of medial and lateral entry pin fixation for the operative treatment of completely displaced extension supracondylar fractures of the humerus in children. METHODS: This prospective, randomized clinical trial had sufficient power to detect a 10% difference in the rate of loss of reduction between the two groups. The techniques of lateral entry and medial and lateral entry pin fixation were standardized in terms of the pin location, the pin size, the incision and position of the elbow used for medial pin placement, and the postoperative course. The primary study end points were a major loss of reduction and iatrogenic ulnar nerve injury. Secondary study end points included radiographic measurements, clinical alignment, Flynn grade, elbow range of motion, function, and complications. RESULTS: The lateral entry group (twenty eight patients) and the medial and lateral entry group (twenty-four patients) were similar in terms of mean age, sex distribution, and preoperative displacement, comminution, and associated neurovascular status. No patient in either group had a major loss of reduction. There was no significant difference between the rates of mild loss of reduction, which occurred in six of the twenty eight patients treated with lateral entry and one of the twenty-four treated with medial and lateral entry (p = 0.107). There were no cases of iatrogenic ulnar nerve injury in either group. There were also no significant differences (p > 0.05) between groups with respect to the Baumann angle, change in the Baumann angle, humerocapitellar angle, change in the humerocapitellar angle, Flynn grade, carrying angle, elbow flexion, elbow extension, total elbow range of motion, return to function, or complications. CONCLUSIONS: With use of the specific techniques employed in this study, both lateral entry pin fixation and medial and lateral entry pin fixation are effective in the treatment of completely displaced (type-III) extension supracondylar fractures of the humerus in children. LEVEL OF EVIDENCE: Therapeutic Level I. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403791 TI - Loss of pin fixation in displaced supracondylar humeral fractures in children: causes and prevention. AB - BACKGROUND: Although the results are generally good following pin fixation of supracondylar humeral fractures in children, occasionally there is postoperative displacement. The purposes of the present study were to identify the causes leading to loss of fixation after pin fixation and to present methods for prevention. METHODS: We evaluated 322 displaced supracondylar humeral fractures that had been treated with percutaneous pin fixation. We examined fracture classification, pin configuration, intraoperative alignment after fixation, change in alignment after fixation, details of additional procedures, and final radiographic and clinical outcomes. RESULTS: Adequate radiographs were available for 279 of the 322 fractures. Eight (2.9%) of the 279 fractures were associated with postoperative loss of fixation; all eight were Gartland type-III fractures. Seven of these eight fractures initially had been treated with two lateral-entry pins, and one had been treated with two crossed pins. In patients with Gartland type-III fractures, loss of fixation was successfully avoided more often when three pins were used (with fixation being maintained in thirty-seven of thirty seven patients) as opposed to when two lateral-entry pins were used (with fixation being maintained in thirty-five of forty-two patients) (p = 0.01). In all cases, loss of fixation was due to technical errors that were identifiable on the intraoperative fluoroscopic images and that could have been prevented with proper technique. We identified three types of pin-fixation errors: (1) failure to engage both fragments with two pins or more, (2) failure to achieve bicortical fixation with two pins or more, and (3) failure to achieve adequate pin separation (>2 mm) at the fracture site. CONCLUSIONS: Postoperative displacement following pin fixation of supracondylar humeral fractures in children is uncommon. In the present series, loss of fixation was most likely to occur when Gartland type-III fractures were treated with two lateral-entry pins. There were no failures when three pins were used. In all cases of failure, there were identifiable technical errors in pin placement. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403792 TI - Fresh stored allografts for the treatment of osteochondral defects of the knee. AB - BACKGROUND: Osteochondral allograft reconstruction of articular cartilage defects is a well-established cartilage repair strategy. Currently, fresh osteochondral allograft tissue is commercially available to clinicians approximately thirty days following graft harvest. Little clinical information is available on the outcome of patients who have been treated with fresh allografts stored for several weeks or more. The purpose of this study was to prospectively analyze the clinical outcome and graft morphology of patients who received fresh, hypothermically stored, allograft tissue for the treatment of symptomatic chondral and osteochondral defects of the knee. METHODS: Nineteen patients with symptomatic chondral and osteochondral lesions of the knee who were treated with fresh osteochondral allografts between 1999 and 2002 were prospectively followed. The mean age at the time of surgery was thirty-four years. Validated outcomes instruments (the Activities of Daily Living Scale and the Short Form-36) were used; all patients must have had a baseline functional score prior to surgery and a minimum two-year follow-up evaluation. All grafts were obtained from commercial vendors; the mean preimplantation storage time of the graft was thirty days (range, seventeen to forty-two days). The mean lesion size was 602 mm(2). Magnetic resonance imaging was used to evaluate the morphologic characteristics of the implanted grafts. RESULTS: The mean duration of clinical follow-up was forty-eight months (range, twenty-one to sixty-eight months). The mean score (and standard deviation) on the Activities of Daily Living Scale increased from a baseline of 56 +/- 24 to 70 +/- 22 at the time of the final follow-up (p < 0.05). The mean Short Form-36 score increased from a baseline of 51 +/- 23 to 66 +/- 24 at the time of final follow-up (p < 0.005). With the numbers studied, we could not correlate graft storage time, body mass index, lesion size, lesion location, or patient age with the functional outcome scores. At a mean follow-up interval of twenty-five months, cartilage-sensitive magnetic resonance imaging demonstrated that the normal articular cartilage thickness was preserved in eighteen implanted grafts, and allograft cartilage signal properties were isointense relative to normal articular cartilage in eight of the eighteen grafts. Osseous trabecular incorporation of the allograft was complete or partial in fourteen patients and poor in four patients. Complete or partial trabecular incorporation positively correlated with Short Form-36 scores at the time of follow-up (r = 0.487, p < 0.05). CONCLUSIONS: Fresh osteochondral allografts that were hypothermically stored between seventeen and forty-two days were effective in the short term both structurally and functionally in reconstructing symptomatic chondral and osteochondral lesions of the knee. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403793 TI - Humeral hemiarthroplasty with biologic resurfacing of the glenoid for glenohumeral arthritis. Two to fifteen-year outcomes. AB - BACKGROUND: Biologic glenoid resurfacing was developed in 1988 as an alternative to total shoulder arthroplasty in selected (usually younger) patients with primary, posttraumatic, or postreconstructive glenohumeral arthritis. A variety of biologic surfaces, including anterior capsule, autogenous fascia lata, and Achilles tendon allograft, have been combined with a humeral hemiarthroplasty. METHODS: From November 1988 to November 2003, thirty-four patients (thirty-six shoulders) who were managed with biologic glenoid resurfacing and humeral head replacement either with cement (ten shoulders) or without cement (twenty-six shoulders) were followed prospectively. The study group included thirty men and four women with an average age of fifty-one years. The diagnoses included primary glenohumeral osteoarthritis (eighteen shoulders), postreconstructive arthritis (twelve), posttraumatic arthritis (five), and osteonecrosis (one). Anterior capsule was used for seven shoulders, autogenous fascia lata for eleven, and Achilles tendon allograft for eighteen. All shoulders were assessed clinically and with serial radiographs. RESULTS: The mean American Shoulder and Elbow Surgeons score was 39 points preoperatively and 91 points at the time of the most recent follow-up. According to Neer's criteria, the result was excellent for eighteen shoulders, satisfactory for thirteen, and unsatisfactory for five. Glenoid erosion averaged 7.2 mm and appeared to stabilize at five years. There were no revisions for humeral component loosening. Complications included infection (two patients), instability (three patients), brachial plexitis (one patient), and deep-vein thrombosis (one patient). Factors that appeared to be associated with unsatisfactory results were the use of capsular tissue as the resurfacing material and infection. CONCLUSIONS: Biologic resurfacing of the glenoid can provide pain relief similar to total shoulder arthroplasty. It allows selected younger patients to maintain an active lifestyle, including weight lifting and manual work, without the risk of polyethylene wear. On the basis of this and previous reviews, we currently recommend Achilles tendon allograft as the preferred resurfacing material when this option is chosen. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403794 TI - Early results of conversion of a failed femoral component in hip resurfacing arthroplasty. AB - BACKGROUND: A theoretical advantage of resurfacing arthroplasty of the hip is that a failed femoral component can be safely and successfully revised to a total hip arthroplasty. To our knowledge, this advantage has not been demonstrated to date. METHODS: Twenty-one metal-on-metal resurfacing arthroplasties in twenty patients with an average age of 50.2 years were converted to a conventional stemmed total hip arthroplasty because of femoral component failure. In eighteen hips, the acetabular component was retained, and in three hips both components were revised. The results in the resurfacing conversion group were compared with those in a group of fifty-eight patients who had undergone sixty-four primary total hip arthroplasties that had been performed during the same time-period by the same surgeon. Clinical evaluations (the Harris hip score, the University of California at Los Angeles pain, walking, and activity scores and the Short Form 12 score) and radiographic evaluations were performed. The average duration of follow-up was forty-six months for the conversion arthroplasty group and fifty seven months for the primary conventional total hip arthroplasty group. RESULTS: There was no significant difference between the conversion arthroplasty group and the conventional arthroplasty group with regard to operative time, blood loss, or complication rates. At the time of the most recent follow-up, with the numbers studied, there were no significant differences between the two groups with regard to the mean Harris hip score; the University of California at Los Angeles pain, walking, and activity score; or the SF-12 score. As assessed radiographically, the quality of component fixation and the alignment of the reconstruction were equivalent between the two groups. There had been no instances of aseptic loosening of the femoral or the acetabular component in either group, and there had been no dislocations after conversion of a resurfacing arthroplasty. CONCLUSIONS: Conversion of a hip resurfacing with a femoral-side failure to a total hip arthroplasty appears to be comparable with primary total hip arthroplasty in terms of surgical effort, safety, and early clinical outcomes. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403795 TI - Detection of orthopaedic implants in vivo by enhanced-sensitivity, walk-through metal detectors. AB - BACKGROUND: Since the September 11, 2001, World Trade Center terrorist attack, airports worldwide have heightened their security standards in efforts to discourage terrorist attacks. Patients have become increasingly concerned about whether their metallic implants will set off airport metal detectors. The purpose of this study was to assess rates of detection of various orthopaedic implants by airport detectors with the new security sensitivities. METHODS: One hundred and twenty-nine volunteers with a total of 149 implants were asked to walk through an M-Scope three-zone metal detector at two sensitivity settings. Low sensitivity was equivalent to the United States Transportation Security Administration setting for regular security, and high sensitivity was equivalent to its standard for high security. RESULTS: Of the 149 implants in 129 patients who were screened, eighty-four (56%) were trauma hardware, including intramedullary nails, plates, screws, and Kirschner wires, and sixty-five (44%) were arthroplasty implants. Seventy-seven (52%) of the 149 implants were detected by the metal detector at one or both settings. Multivariate analysis revealed that the type (p < 0.001), material (p < 0.001), and location (p < 0.001) of the implant were independent predictors of detection. The overall rate of detection was 88% for prosthetic replacements compared with 32% for plates, with the likelihood of detection being fifteen times greater (odds ratio = 15.0, 95% confidence interval = 5.9 to 39.1) for the prosthetic replacements. All total hip replacements and 90% of the total knee replacements were detected at the low-sensitivity setting. Intramedullary nails and Kirschner wires were not detected. The overall detection rate was 67% for implants in the lower extremity, 17% for those in the upper extremity, and 14% for those in the spine. The detection rate for implants in the lower extremity was ten times higher than that for implants in the upper extremity and eleven times higher than that for implants in the spine. CONCLUSIONS: More than half of all orthopaedic implants may be detected by metal detectors used at commercial airports. Total joint prostheses will routinely set off the detector, whereas nails, plates, screws, and wires are rarely detected. Cobalt-chromium and titanium implants are more likely to be detected than stainless-steel implants. PMID- 17403796 TI - Isolated arthroscopic biceps tenotomy or tenodesis improves symptoms in patients with massive irreparable rotator cuff tears. AB - BACKGROUND: Lesions of the long head of the biceps tendon are often associated with massive rotator cuff tears and may be responsible for shoulder pain and dysfunction. The purpose of this study was to evaluate the clinical and radiographic outcomes of isolated arthroscopic biceps tenotomy or tenodesis as treatment for persistent shoulder pain and dysfunction due to an irreparable rotator cuff tear associated with a biceps lesion. METHODS: We conducted a retrospective study of sixty-eight consecutive patients (mean age [and standard deviation], 68 +/- 6 years) in whom a total of seventy-two irreparable rotator cuff tears had been treated arthroscopically with biceps tenotomy or tenodesis. A simple tenotomy was performed in thirty-nine cases, and a tenodesis was performed in thirty-three. No associated acromioplasty was performed. All patients were evaluated clinically and radiographically by an independent observer at a mean of thirty-five months postoperatively. RESULTS: Fifty-three patients (78%) were satisfied with the result. The mean Constant score improved from 46.3 +/- 11.9 points preoperatively to 66.5 +/- 16.3 points postoperatively (p < 0.001). A healthy-appearing teres minor on preoperative imaging was associated with significantly increased postoperative external rotation (40.4 degrees +/- 19.8 degrees compared with 18.1 degrees +/- 18.4 degrees ) and a significantly higher Constant score (p < 0.05 for both) compared with the values for the patients with an absent or atrophic teres minor preoperatively. Three patients with pseudoparalysis of the shoulder did not benefit from the procedure and did not regain active elevation above the horizontal level. In contrast, the fifteen patients with painful loss of active elevation recovered active elevation. The acromiohumeral distance decreased 1.1 +/- 1.9 mm on the average, and glenohumeral osteoarthritis developed in only one patient. The results did not differ between the tenotomy and tenodesis groups (mean Constant score, 61.2 +/- 18 points and 72.8 +/- 12 points, respectively). The "Popeye" sign was clinically apparent in twenty-four (62%) of the shoulders that had been treated with a tenotomy; of the sixteen patients who noticed it, none were bothered by it. CONCLUSIONS: Both arthroscopic biceps tenotomy and arthroscopic biceps tenodesis can effectively treat severe pain or dysfunction caused by an irreparable rotator cuff tear associated with a biceps lesion. Shoulder function is significantly inferior if the teres minor is atrophic or absent. Pseudoparalysis of the shoulder and severe rotator cuff arthropathy are contraindications to this procedure. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403797 TI - The anatomy of the posterior aspect of the knee. An anatomic study. AB - BACKGROUND: The orthopaedic literature contains relatively little quantitative information regarding the anatomy of the posterior aspect of the knee. The purpose of the present study was to provide a detailed description of, and to propose a standard nomenclature for, the anatomy of the posterior aspect of the knee. METHODS: Detailed dissection of twenty nonpaired, fresh-frozen knees was performed. Posterior knee structures were measured according to length, width, and/or distance to reproducible osseous landmarks. RESULTS: The semimembranosus tendon had eight attachments distal to the main common tendon. The main components were a lateral expansion to the oblique popliteal ligament; a direct arm, which attached to the tibia; and an anterior arm. The oblique popliteal ligament, the largest posterior knee structure, formed a broad fascial sheath over the posterior aspect of the knee and measured 48.0 mm in length and 9.5 mm wide at its medial origin and 16.4 mm wide at its lateral attachment. It had two lateral attachments, one to the meniscofemoral portion of the posterolateral joint capsule and one to the tibia, along the lateral border of the posterior cruciate ligament facet. The semimembranosus also had a distal tibial expansion, which formed a posterior fascial layer over the popliteus muscle. A thickening of the posterior joint capsule, the proximal popliteus capsular expansion, which in this study averaged 40.5 mm in length, connected the posteromedial knee capsule at its attachment at the intercondylar notch to the medial border of the popliteus musculotendinous junction. The plantaris muscle, popliteofibular ligament, fabellofibular ligament, and semimembranosus bursa were present in all specimens. CONCLUSIONS: The anatomy of the posterior aspect of the knee is quite complex. This study provides information that can lead to further biomechanical, radiographic imaging, and clinical studies of the importance of these posterior knee structures. PMID- 17403798 TI - The osteoporosis self-assessment screening tool: a useful tool for the orthopaedic surgeon. AB - BACKGROUND: Simple and effective methods are needed to identify patients at risk for osteoporosis or osteoporosis-related fracture so that they can be screened with use of dual x-ray absorptiometry and counseled for treatment. Currently, we use a cumbersome survey assessing thirty-two risk factors. A much simpler score based on the Osteoporosis Self-Assessment Screening Tool (OST score) has been established as highly sensitive and specific in women, but similar data are lacking for men. This score is calculated by subtracting the age of the patient in years from the weight in kilograms and multiplying the result by 0.2. Our goal was to test the hypothesis that the OST score is more sensitive and specific than our extensive risk-assessment survey in men. METHODS: Using axial dual x-ray absorptiometry analysis, we evaluated a cohort of men who had either responded to our newspaper advertisement or were seen as patients in our orthopaedic clinic. Patients filled out the risk-assessment survey at the time of scanning. Osteoporosis was defined as a T-score of -2.5 or less in the lumbar spine, hip, or femoral neck. RESULTS: Twenty-seven (17%) of 158 white men, with a mean age of 67.5 years and a mean weight of 85.3 kg, had osteoporosis. After analysis of the thirty-two risk factors, two remained as significant independent predictors in the final multivariable model (p = 0.042 and p = 0.015). This model had an area under the receiver operating characteristic curve of 0.68 (>0.70 is considered to provide acceptable discrimination). The OST scores ranged from -6 (greatest risk) to 16 (least risk). With use of the OST score to predict osteoporosis, the area under the receiver operating characteristic curve was 0.76. The cutoff of an OST score of <2 provided the largest area under the receiver operating characteristic curve (0.74), with test characteristics for an OST score of <2 including a sensitivity of 85%, specificity of 64%, positive predictive value of 31%, and negative predictive value of 96%. CONCLUSIONS: The Osteoporosis Self-Assessment Screening Tool score is superior to a broad risk-factor analysis in the identification of men at risk for osteoporosis or osteoporotic fractures. We have found it simple to use in our clinic to determine which patients should undergo dual x-ray absorptiometry screening. LEVEL OF EVIDENCE: Diagnostic Level I. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403799 TI - Quality of life following femoral head-neck osteochondroplasty for femoroacetabular impingement. AB - BACKGROUND: Femoroacetabular impingement has been recently described as a common cause of hip pain and labral tears in young adults. We evaluated the early clinical results and quality of life after osteochondroplasty of the femoral head neck junction for the treatment of femoroacetabular impingement. METHODS: Thirty seven hips in thirty-four patients with persistent hip pain and a mean age of 40.5 years underwent surgical dislocation of the hip and osteochondroplasty of the femoral head-neck junction for the treatment of camtype femoroacetabular impingement. All of the patients had had preoperative evidence of pathological changes in the labrum on imaging. The clinical course and the quality of life were assessed postoperatively. RESULTS: The mean score on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) increased from 61.2 points preoperatively to 81.4 points at a mean of 3.1 years postoperatively (p < 0.001), the mean University of California at Los Angeles (UCLA) activity score increased from 4.8 to 7.5 points (p < 0.001), the mean Short Form-12 (SF-12) physical component score increased from 37.3 to 45.6 points (p < 0.001), and the mean SF 12 mental component score increased from 46.4 to 51.2 points (p = 0.031). None of the hips underwent additional reconstructive surgery. There were no cases of osteonecrosis. Nine patients underwent screw removal from the greater trochanter because of persistent bursitis. Six of the thirty-four patients were dissatisfied with the outcome. CONCLUSIONS: Cam-type femoroacetabular impingement is associated with insufficient concavity at the anterolateral head-neck junction and with pathological changes in the labrum. Osteochondroplasty of the femoral head-neck junction following surgical dislocation of the hip joint is safe and effective and can provide a significant improvement in the overall quality of life of most patients. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403800 TI - Projections of primary and revision hip and knee arthroplasty in the United States from 2005 to 2030. AB - BACKGROUND: Over the past decade, there has been an increase in the number of revision total hip and knee arthroplasties performed in the United States. The purpose of this study was to formulate projections for the number of primary and revision total hip and knee arthroplasties that will be performed in the United States through 2030. METHODS: The Nationwide Inpatient Sample (1990 to 2003) was used in conjunction with United States Census Bureau data to quantify primary and revision arthroplasty rates as a function of age, gender, race and/or ethnicity, and census region. Projections were performed with use of Poisson regression on historical procedure rates in combination with population projections from 2005 to 2030. RESULTS: By 2030, the demand for primary total hip arthroplasties is estimated to grow by 174% to 572,000. The demand for primary total knee arthroplasties is projected to grow by 673% to 3.48 million procedures. The demand for hip revision procedures is projected to double by the year 2026, while the demand for knee revisions is expected to double by 2015. Although hip revisions are currently more frequently performed than knee revisions, the demand for knee revisions is expected to surpass the demand for hip revisions after 2007. Overall, total hip and total knee revisions are projected to grow by 137% and 601%, respectively, between 2005 and 2030. CONCLUSIONS: These large projected increases in demand for total hip and knee arthroplasties provide a quantitative basis for future policy decisions related to the numbers of orthopaedic surgeons needed to perform these procedures and the deployment of appropriate resources to serve this need. PMID- 17403801 TI - Restore orthobiologic implant: not recommended for augmentation of rotator cuff repairs. AB - BACKGROUND: Following repairs of large-to-massive tears of the rotator cuff, the rates of tendon retears are high and often involve tissue deficiency. Animal studies of the Restore Orthobiologic Implant, a collagen-based material derived from the small intestine mucosa of pigs, have indicated that it might be used to help overcome such problems. We carried out a study to determine whether patients who received this xenograft to augment a rotator cuff repair exhibited greater shoulder strength, shoulder function, and/or resistance to retearing. METHODS: We compared data from a group of patients who had undergone conventional rotator cuff repair with xenograft augmentation (the xenograft group) with data from a group in whom a repair had been done by the same surgeon without augmentation (the controls). The groups were matched for gender, mean age, and mean size of the rotator cuff tear. All subjects completed a pain and function questionnaire and were given a systematic clinical shoulder examination preoperatively and at three, six, and twenty-four months postoperatively. The twenty-four-month visit included magnetic resonance imaging to determine whether a retear had occurred. RESULTS: Four patients who had received a xenograft had a severe postoperative reaction requiring surgical treatment. At two years after the surgery, six of the ten tendons repaired with a xenograft and seven of the twelve control tendons had retorn, as documented by magnetic resonance imaging. The patients with a xenograft had significantly less lift-off strength, as measured with a dynamometer, and significantly less strength in internal rotation and adduction than the controls at two years after the surgery (all p < 0.05). Also, the xenograft group had significantly more impingement in external rotation, a slower rate of resolution of pain during activities, more difficulty with hand-behind the-back activities, and less sports participation (all p < 0.05). CONCLUSIONS: Two years after surgical repair of a large rotator cuff defect supplemented with a xenograft, patients had several persisting deficits and no recognizable benefit as compared with the results in a control group. In view of these findings, together with the unsatisfactorily high proportion of patients with a severe inflammatory reaction to the xenograft, we do not recommend use of the Restore Orthobiologic Implant in its present form. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403802 TI - In vivo efficacy of antimicrobial-coated devices. AB - BACKGROUND: Since device colonization is a prelude to infection, an antimicrobial coated device that reduces bacterial colonization can potentially protect against infection. The objective of this animal study was to assess the efficacy of a coating with minocycline and rifampin to prevent colonization of a grit-blasted titanium implant and subsequent osteomyelitis. METHODS: Twenty-five rabbits underwent implantation of a titanium-alloy pin, either coated with minocycline and rifampin (thirteen rabbits) or uncoated (twelve rabbits), into the right femoral medullary canal. The implanted devices were inoculated with 500 CFU (colony-forming units) of Staphylococcus aureus prior to wound closure. The rabbits were killed one week later, and the removed device, femoral bone, a specimen obtained by swabbing the track surrounding the device, and blood were cultured. The rates of device colonization, osteomyelitis, and device-related osteomyelitis were compared between the two groups of rabbits. RESULTS: The antimicrobial-coated devices had a significantly lower rate of colonization than the uncoated devices (five of thirteen compared with twelve of twelve, p = 0.0016) and were associated with significantly lower rates of osteomyelitis (six of thirteen compared with twelve of twelve, p = 0.005) and device-related osteomyelitis (five of thirteen compared with twelve of twelve, p = 0.0016). Bacteremia did not develop in any rabbit. CONCLUSIONS: Orthopaedic devices coated with minocycline and rifampin significantly protected against device colonization and infection due to Staphylococcus aureus in this in vivo rabbit model. CLINICAL RELEVANCE: It is possible that orthopaedic devices coated with this unique combination of antimicrobial agents may protect against the development of clinical infection in humans. PMID- 17403803 TI - Uncemented total hip arthroplasty in patients with a history of pelvic irradiation for prostate cancer. AB - BACKGROUND: Pelvic irradiation for a malignant tumor may cause osteonecrosis of the acetabulum. The purpose of this study was to evaluate the outcome of uncemented total hip arthroplasty in patients with a history of pelvic irradiation for the treatment of prostate cancer. METHODS: We performed a retrospective review of the clinical records and radiographs of fifty-eight patients (sixty-six hips) who had had radiation therapy for prostate cancer and had subsequently undergone an elective primary uncemented total hip arthroplasty at our institution. The mean age of the patients at the time of the index operation was seventy-four years. The mean duration of follow-up was 4.8 years (range, two to 7.5 years). RESULTS: At the time of the final follow-up, fifty-one patients (fifty-eight hips) who were still living and had been followed for a minimum of two years had a well-ingrown and functioning replacement. The mean Harris hip score had significantly improved from 47 points preoperatively to 90 points at the time of the final follow-up (p < 0.05). The mean scores on the physical and mental health measures of the Short Form-36 had also improved significantly from 45.1 and 65.3 points, respectively, before the operation to 73.4 and 83.7 points postoperatively (p < 0.05 for both). There was no aseptic loosening of either component in any of the hips. Two hips had revision of the femoral component; one was revised because of a periprosthetic fracture of the femur and the other because of subsidence of the femoral component. CONCLUSIONS: Uncemented total hip arthroplasty can be a successful option for the treatment of coxarthrosis in patients with a history of pelvic irradiation for prostate cancer. Osseointegration of uncemented components does not seem to be compromised in these patients in the short term. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403804 TI - Impact of ankle-foot orthoses on static foot alignment in children with cerebral palsy. AB - BACKGROUND: Children with cerebral palsy who are able to walk are often managed with an ankle-foot orthosis to assist with walking. Previous studies have shown kinematic, kinetic, and energetic benefits during gait with the addition of an ankle-foot orthosis, although the mechanism of this gait improvement is unknown. The ability of orthoses to correct foot malalignment in children with cerebral palsy is not known. The current study was performed to determine the impact of orthoses on static foot alignment in children with cerebral palsy. METHODS: A retrospective radiographic review was performed for 160 feet (102 patients). All patients had a diagnosis of cerebral palsy. Standing anteroposterior and lateral radiographs of the foot and ankle were made with the patient barefoot and while wearing the prescribed orthosis and were compared with use of the technique of quantitative segmental analysis of foot and ankle alignment. RESULTS: Analysis of the foot and ankle radiographs made with the patient barefoot and while wearing the brace revealed significant changes in all measurements of segmental alignment (p < 0.05). The magnitudes of these differences were small (<6 degrees or <10%) and would be considered clinically unimportant. The coupled malalignment of equinoplanovalgus (clinical flatfoot) showed radiographic correction of at least one segment (hindfoot, midfoot, or forefoot) to within the normal range in 24% to 44% of the feet. The coupled malalignment of equinocavovarus (clinical high arched foot) showed correction of at least one segment to within the normal range in 5% to 20% of feet. CONCLUSIONS: The present study demonstrates that the use of the ankle-foot orthoses failed to improve the static foot alignment in the majority of feet in children with cerebral palsy who were able to walk. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403805 TI - The twenty to twenty-five-year outcomes of the Harris design-2 matte-finished cemented total hip replacement. A concise follow-up of a previous report. AB - We previously reported the ten to fifteen-year results for 195 matte-finished Harris Design-2 total hip replacements that had been inserted with cement by two surgeons in 166 patients with osteoarthritis. The purpose of the present report is to update that study and report the twenty to twenty-five-year outcomes. The patients were followed prospectively on the basis of clinical assessment with use of the Harris hip score and radiographic analysis. One hundred and forty-nine patients (90%) had a functioning implant at the time of death or, if living, at twenty to twenty-five years of follow-up. A total of ten patients (ten hips; 5%) underwent a revision because of aseptic loosening of the acetabular component (two hips; 1%), the femoral component (four hips; 2%), or both components (four hips; 2%). At twenty-five years, Kaplan-Meier analysis revealed a survival rate of 83% +/- 6% with revision for any reason as the end point. The survival rate was 86% +/- 6% for the femoral component and 93% +/- 3% for the acetabular component with aseptic loosening as the end point. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403806 TI - The cost-effectiveness of extended-duration antithrombotic prophylaxis after total hip arthroplasty. AB - BACKGROUND: Although the risk of thromboembolism after total hip arthroplasty continues beyond hospital discharge, the cost-effectiveness of extending prophylaxis beyond hospitalization is unclear. We compared the cost-effectiveness of an extended duration of antithrombotic prophylaxis following total hip arthroplasty, with use of low-molecular-weight heparin or warfarin administered for twenty-eight days beyond hospital discharge, in terms of incremental cost per quality-adjusted life year gained. METHODS: The economic analysis was structured around a decision tree characterizing the consequences of extended prophylaxis choices following total hip arthroplasty. The health benefits of extended antithrombotic prophylaxis, measured as the reduction in symptomatic venous thromboembolic events and deaths for each treatment alternative, were determined through a systematic review of the literature. Gains in quality-adjusted life years were based on the distribution of life years remaining for all patients undergoing total hip arthroplasty in Canada in 2003, weighted by utilities derived from the literature. The cost analysis, in 2006 Canadian dollars, took a direct payer perspective with a ninety-day time horizon. RESULTS: There was a net gain in quality-adjusted life years in both cohorts that received extended prophylaxis relative to the cohort that received no extended prophylaxis (7.5 quality-adjusted life years per 1000 patients treated with low-molecular-weight heparin and 5.5 quality-adjusted life years per 1000 patients treated with warfarin), although these gains were not significant. The net treatment costs per 1000 patients treated were $799,104 with low-molecular-weight heparin and $72,236 with warfarin. In comparison with the cohort that received no extended prophylaxis, the cost-effectiveness of low-molecular-weight heparin was $106,454 per quality-adjusted life year gained and the cost-effectiveness of warfarin was $13,115 per quality-adjusted life year gained. CONCLUSIONS: There is insufficient economic evidence to support extended thromboprophylaxis with low-molecular weight heparin following total hip arthroplasty. Although the cost-effectiveness of warfarin was potentially quite favorable, this finding was based on limited clinical evidence. Further research is required to clarify the benefits of extended prophylaxis, particularly with warfarin. LEVEL OF EVIDENCE: Economic and decision analysis, Level I. See Instructions to Authors for a complete description of levels of evidence. PMID- 17403807 TI - Risk of superior gluteal nerve and gluteus medius muscle injury during femoral nail insertion. AB - BACKGROUND: Abduction weakness and limping is a well-recognized complication of closed antegrade insertion of femoral nails. Iatrogenic injuries to the superior gluteal nerve and the gluteus medius muscle are the most likely contributing factors. The purpose of this study of cadavers was to assess the risk of nerve and muscle injury with various lower-limb positions used during nail insertion. METHODS: We studied thirteen hips of ten formalin-fixed adult cadavers. With the cadaver in the full lateral position, a 9-mm reamer was introduced in a retrograde fashion from the intercondylar notch and passed through the gluteus medius muscle. The distance between the point of entry of the reamer into the undersurface of this muscle and the inferior main branch of the superior gluteal nerve (the nerve-reamer distance) and the distance between the entry and exit points of the reamer in the gluteus medius muscle (the intramuscle distance) were measured in three different hip positions: 15 degrees of flexion and 15 degrees of adduction (Position 1), 30 degrees of flexion and 30 degrees of adduction (Position 2), and 60 degrees of flexion and 30 degrees of adduction (Position 3). RESULTS: In Position 1, the average nerve-reamer distance was 7 mm and the average intramuscle distance was 24 mm. In three hips the reamer injured the nerve directly, and in two other hips the distance was or = 1 health screening measure checked in the past year compared with 73.1% of children without special health care needs. Receipt of > or = 1 topic of anticipatory guidance was reported for 69.8% of children with special health care needs compared with 55.2% of children without special health care needs. Black and Hispanic caregivers of children with special health care needs were more likely than others to report receipt of all 6 categories of anticipatory guidance measured in this study. CONCLUSIONS: We found that caregivers of children with special health care needs were more likely to report receipt of anticipatory guidance and health screening than were caregivers of children without special health care needs. Although a majority of these caregivers reported receiving some health screening and anticipatory guidance on an annual basis, there are clear gaps in the delivery of preventive health services. This study identifies areas for improvement in the delivery of preventive health services for children with special health care needs and children in general. PMID- 17403826 TI - Association between parental depression and children's health care use. AB - OBJECTIVE: The objective of this study was to determine the association between parental depression and pediatric health care use patterns. METHODS: We selected all children who were 0 to 17 years of age, enrolled in Kaiser Permanente of Colorado during the study period July 1997 to December 2002, and linked to at least 1 parent/subscriber who was enrolled for at least 6 months during that period. Unexposed children were selected from a pool of children whose parents did not have a depression diagnosis. Outcome measures were derived from the child's payment files and electronic medical charts and included 5 categories of use: well-child-care visits, sick visits to primary care departments, specialty clinic visits, emergency department visits, and inpatient visits. We compared the rate of use per enrollment month for these 5 categories between exposed and unexposed children within each of the 5 age strata. RESULTS: Our study population had 24,391 exposed and 45,274 age-matched, unexposed children. For the outcome of well-child-care visits, teenagers showed decreased rates of visits among exposed children. The rate of specialty department visits was higher in exposed children in the 4 oldest age groups. The rates of both emergency department visits and sick visits to primary care departments were higher for exposed children across all 5 age categories. The rate of inpatient visits was higher among exposed children in 2 of the 5 age groups. CONCLUSIONS: Overall, having at least 1 depressed parent is associated with greater rate of emergency department and sick visits across all age groups, greater use of inpatient and specialty services in some age groups, and a lower rate of well-child-care visits among 13- to 17-year olds. This pattern of increased use of expensive resources and decreased use of preventive services represents one of the hidden costs of adult depression. PMID- 17403827 TI - Breastfeeding and hospitalization for diarrheal and respiratory infection in the United Kingdom Millennium Cohort Study. AB - OBJECTIVE: The objective of this study was to measure the effect of breastfeeding on hospitalization for diarrheal and lower respiratory tract infections in the first 8 months after birth in contemporary United Kingdom. METHODS: The study was a population-based survey (sweep 1 of the United Kingdom Millennium Cohort Study). Data on infant feeding, infant health, and a range of confounding factors were available for 15,890 healthy, singleton, term infants who were born in 2000 2002. The main outcome measures were parental report of hospitalization for diarrhea and lower respiratory tract infection in the first 8 months after birth. RESULTS: Seventy percent of infants were breastfed (ever), 34% received breast milk for at least 4 months, and 1.2% were exclusively breastfed for at least 6 months. By 8 months of age, 12% of infants had been hospitalized (1.1% for diarrhea and 3.2% for lower respiratory tract infection). Data analyzed by month of age, with adjustment for confounders, show that exclusive breastfeeding, compared with not breastfeeding, protects against hospitalization for diarrhea and lower respiratory tract infection. The effect of partial breastfeeding is weaker. Population-attributable fractions suggest that an estimated 53% of diarrhea hospitalizations could have been prevented each month by exclusive breastfeeding and 31% by partial breastfeeding. Similarly, 27% of lower respiratory tract infection hospitalizations could have been prevented each month by exclusive breastfeeding and 25% by partial breastfeeding. The protective effect of breastfeeding for these outcomes wears off soon after breastfeeding cessation. CONCLUSIONS: Breastfeeding, particularly when exclusive and prolonged, protects against severe morbidity in contemporary United Kingdom. A population level increase in exclusive, prolonged breastfeeding would be of considerable potential benefit for public health. PMID- 17403828 TI - Results of random drug testing in an adolescent substance abuse program. AB - OBJECTIVE: The objective of this study was to estimate from a random urine drug testing program for adolescents the proportion of drug tests that are susceptible to interpretation errors. METHODS: This was a secondary analysis of a clinical database and chart review from an adolescent outpatient substance abuse program at a large children's hospital. We analyzed from 110 adolescent patients (13-21 years of age) all 710 urine drug test results that were collected between December 2002 and July 2005 and 85 original laboratory reports for tests that were collected between December 2002 and May 2006 and were confirmed positive for opioids. We calculated the percentage of tests that were too dilute to interpret (potential false-negatives) and the percentage of confirmed positive tests for oxycodone that did not result in a positive initial screen (potential false negatives). We also reviewed clinical information to determine whether confirmed positive tests resulted from legitimate use of prescription or over-the-counter medication (potential false-positives). RESULTS: Of 710 drug tests, 40 negative tests were too dilute to interpret properly, and 45 of 217 positive tests resulted from prescription medication use for a total of 85 tests that were susceptible to error. Of the 85 confirmatory laboratory reports reviewed, 43 were positive for oxycodone, but only 16 of these had produced a positive opiate screen. CONCLUSIONS: Unless proper procedures are used in collecting, analyzing, and interpreting laboratory testing for drugs, there is a substantial risk for error. PMID- 17403829 TI - Improving informed consent: suggestions from parents of children with leukemia. AB - OBJECTIVE: The objective of this study was to report suggestions for improving the informed consent process from the perspective of parents of children with leukemia. METHODS: Recommendations for improving informed consent were elicited from 140 parents of children who had been offered participation in a randomized clinical trial for the treatment of their acute leukemia. Four different methods and data collection time points were used with this group of parents, including open-ended, in-person interviews within 72 hours after the informed consent conference; follow-up telephone interviews 6 months after diagnosis; focus groups during year 3 of the project; and a parent advisory group on informed consent meeting in year 4. RESULTS: The most frequently cited suggestions for improving informed consent during the interviews and focus groups related to giving parents more time to make their decision, the amount and type of information provided, organization of the consent conference, communication style, and providing additional materials. During the parent advisory group on informed consent meeting, parents developed specific guidelines for organization of the information that is presented during the consent process that include 7 major components: timing, sequence, checklist, checking for understanding, anticipatory guidance, segue into randomized clinical trial discussion with historical perspective, and choice. CONCLUSIONS: Through the incorporation of parental perspectives that provide an authentic stakeholder voice, our research represents a true partnership approach to improving the consent process. Parents provided practical advice for improving informed consent that can be applied to most adult and pediatric patient populations. PMID- 17403830 TI - Twenty-year trends in fatal injuries to very young children: the persistence of racial disparities. AB - OBJECTIVE: Mortality trends across modifiable injury mechanisms may reflect how well effective injury prevention efforts are penetrating high-risk populations. This study examined all-cause, unintentional, and intentional injury-related mortality in children who were aged 0 to 4 years for evidence of and to quantify racial disparities by injury mechanism. METHODS: Injury analyses used national vital statistics data from January 1, 1981, to December 31, 2003, that were available from the Centers for Disease Control and Prevention. Rate calculations and chi2 test for trends (Mantel extension) used data that were collapsed into 3 year intervals to produce cell sizes with stable estimates. Percentage change for mortality rate ratios used the earliest (1981-1983) and the latest (2001-2003) study period for black, American Indian/Alaskan Native, and Asian/Pacific Islander children, with white children as the comparison group. RESULTS: All cause injury rates declined during the study period, but current mortality ratios for all-cause injury remained higher in black and American Indian/Alaskan Native children and lower in Asian/Pacific Islander children compared with white children. Trend analyses within racial groups demonstrate significant improvements in all groups for unintentional but not intentional injury. Black and American Indian/Alaskan Native children had higher injury risk as a result of residential fire, suffocation, poisoning, falls, motor vehicle traffic, and firearms. Disparities narrowed for residential fire, pedestrian, and poisoning and widened for motor vehicle occupant, unspecified motor vehicle, and suffocation for black and American Indian/Alaskan Native children. CONCLUSIONS: These findings identify injury areas in which disparities narrowed, improvement occurred with maintenance or widening of disparities, and little or no progress was evident. This study further suggests specific mechanisms whereby new strategies and approaches to address areas that are recalcitrant to improvement in absolute rates and/or narrowing of disparities are needed and where increased dissemination of proven efficacious injury prevention efforts to high-risk populations are indicated. PMID- 17403831 TI - Impact of the State Children's Health Insurance Program on adolescents in New York. AB - OBJECTIVES: Adolescents face financial and nonfinancial barriers to health care. Little is known about the impact of health insurance on health care for adolescents. We assessed the impact of New York's State Children's Health Insurance Program on access, use, and quality of care for adolescents. METHODS: Adolescents and their parents from a stratified random sample of new enrollees in New York's State Children's Health Insurance Program were interviewed by telephone shortly after enrollment (baseline, n = 1118 adolescents and their parents) and 1 year later (follow-up, n = 970). Outcome measures included access (having a usual source of care and reported unmet health needs), use (preventive care and other types of visits), and quality (satisfaction with care, receipt of confidential care and preventive counseling). Outcomes were assessed at baseline (year before the State Children's Health Insurance Program) versus follow-up (year during the State Children's Health Insurance Program). RESULTS: The proportion of adolescents who reported having a usual source of care increased during State Children's Health Insurance Program compared with before (69.9% to 87.1%). The proportion with any unmet health care need (54.3% to 42.1%) or with unmet need for preventive care (53.8% to 40.6%) decreased, with elimination of racial disparities that existed before the State Children's Health Insurance Program. After enrollment in the State Children's Health Insurance Program, more adolescents reported having had a preventive care visit (65.9% to 74.2%); emergency department use did not change. No differences in satisfaction were noted, although significant increases were noted in both parent- and adolescent reported rates of having received confidential care and preventive counseling. CONCLUSIONS: Adolescents who enrolled in New York's State Children's Health Insurance Program experienced improved access, use, and quality of care. These findings suggest that the provision of health insurance can help to improve health care for adolescents. PMID- 17403832 TI - Persistent socioeconomic disparities in infant, neonatal, and postneonatal mortality rates in the United States, 1969-2001. AB - OBJECTIVES: This study examines changing patterns of inequalities in US infant, neonatal, and postneonatal mortality rates between 1969 and 2001 by area deprivation and maternal education. METHODS: A deprivation index was linked to county vital records data to derive annual infant mortality rates by deprivation quintiles from 1969 to 2000. Rates by maternal education were computed for 1986, 1991, 1996, and 2001 using national linked birth/infant death files. Log-binomial regression was used to estimate relative risks of infant mortality by deprivation and time period. Cox regression was used to model overall and birth weight specific infant mortality risks by maternal education after adjusting for covariates. Temporal disparities were summarized by log-linear regression and inequality indices. RESULTS: Although absolute disparities have narrowed over time, relative socioeconomic disparities in infant mortality have increased since 1985. In 1985-1989, infants in the most deprived group had, respectively, 36% and 57% higher risks of neonatal and postneonatal mortality than infants in the least deprived group. The corresponding relative risks increased to 43% and 96% in 1995 2000. The adjusted risk of infant mortality was 22% higher in 1986 for mothers with < 12 years of education than for those with > or = 16 years of education, with the relative risk increasing to 41% in 2001. Disparities were greatest among normal birth weight infants, with education-specific relative risks of neonatal and postneonatal mortality increasing significantly between 1986 and 2001. CONCLUSIONS: Dramatic declines in infant mortality among all of the socioeconomic groups during 1969-2001 represent a major public health success. However, substantial socioeconomic disparities persisted in both neonatal and postneonatal mortality. Relatively larger declines in infant and postneonatal mortality among higher socioeconomic groups have contributed to the widening gap in mortality since 1985. Persistent disparities in infant mortality may reflect increasing polarization among socioeconomic groups in material and social conditions, smoking during pregnancy, and health care services. PMID- 17403833 TI - Injury-prevention counseling and behavior among US children: results from the second Injury Control and Risk Survey. AB - OBJECTIVES: The purpose of this work was to provide recent national prevalence estimates of pediatric injury-prevention counseling by health care providers, to compare these latest findings with those from a similar survey conducted in 1994, and to ascertain the association between counseling and safety behaviors. METHODS: We conducted a cross-sectional, list-assisted random-digit-dial telephone survey of randomly selected children in English- or Spanish-speaking households in all 50 US states and the District of Columbia. The main outcome measures were respondents' reports that they or their children received injury prevention counseling from their child's health care provider in the 12 months preceding the interview, children's practices of safety behaviors, and the association of injury-prevention counseling and such behaviors. RESULTS: The overall proportion of US children receiving any injury-prevention counseling (42.4%) remained relatively unchanged, whereas counseling on selected injury prevention topics increased significantly compared with reports based on the 1994 survey. Topic-specific injury-prevention counseling was positively associated with the posting of the poison control center telephone number in homes with children <6 years of age and with bicycle-helmet use among children 5 to 14 years of age. CONCLUSIONS: Although the prevalence of pediatric injury-prevention counseling remains low, such counseling was associated with safer behaviors. This suggests the importance of pediatric injury-prevention counseling and indicates the need for health care providers to increase pediatric injury-prevention counseling in clinical practices. PMID- 17403834 TI - Masculine beliefs, parental communication, and male adolescents' health care use. AB - OBJECTIVES: Male adolescents frequently become disconnected from health care, especially as they get older, which limits physicians' abilities to address their health needs and results in missed opportunities to connect them to the health care system as they enter adulthood. In this study we tested the ability of modifiable (beliefs about masculinity, parental communication, sex education, and health insurance) and nonmodifiable (age, race/ethnicity, and region of residence) factors to prospectively predict health care use by male adolescents. PATIENTS AND METHODS: We conducted a prospective analysis of data from 1677 male participants aged 15 to 19 years who completed the National Survey of Adolescent Males, a household probability survey conducted throughout the United States in 1988 (wave 1, participation rate: 74%) and in 1990-1991 (wave 2, follow-up rate: 89%). We present percentages and adjusted relative risks of the factors that predict male adolescents' self-report of a physical examination by a regular provider in the past year measured at wave 2. RESULTS: On average, 1067 (66%) of 1677 male adolescents at wave 2 reported having a physical examination within the last year. Factors associated with a lower likelihood of a physical examination included living in the South, Midwest, and West; being older in age; and holding more traditional masculine beliefs. Factors associated with a higher likelihood of a physical examination included communicating about reproductive health with both parents and being insured. Male adolescents who were sexually active or engaged in > or = 2 other risk behaviors had neither a higher nor lower likelihood of a physical examination. CONCLUSIONS: Efforts to enhance male adolescents' health through health care should include work to modify masculine stereotypes, improve mothers' and fathers' communication about health with their sons, expand health insurance coverage, and identify interventions to connect male adolescents at increased risk for health problems with health care. PMID- 17403835 TI - Can juvenile justice detention facilities meet the call of the American Academy of Pediatrics and National Commission on Correctional Health Care? A national analysis of current practices. AB - OBJECTIVE: Despite the recommendation of the American Academy of Pediatrics, just 53 of the approximately 3500 juvenile justice residential facilities in the United States have received voluntary accreditation for facility health care from the National Commission on Correctional Health Care. This suggests either that facilities do not meet the standards of care or do not seek accreditation. This study describes whether and under what conditions juvenile detention facilities (a narrowly defined subset of all facility types) adhere to some of the standards outlined by the National Commission on Correctional Health Care and promoted by the American Academy of Pediatrics. METHODS: Data from 2 national censuses of juvenile justice residential facilities (n = 726) were used to describe detention facility performance in terms of 10 types of service provision, ranging from health screening to communicable-disease testing. Multivariate models predicting high levels of service provision were estimated. RESULTS: Juvenile detention centers partially meet some of the minimum standards. Most services can be garnered at some level; however, they tend to be provided on an ad hoc basis for portions of the population rather than systematically for the whole population. Detention centers most likely to provide a higher tier of services tend to be those that have longer average lengths of stay, are larger, and are government owned. There are also geographic and racial differences in quality and scope of health services. CONCLUSIONS: Juvenile facilities have been provided a single set of standards for a diverse system with tremendous variation across and within facility types. Detention centers are just one specialized type. Very few detention centers meet a minimum standard of care, which suggests that standards are simply not being met (hence the low levels of accreditation). The findings of this study call into question whether detention facilities with little in the way of health care infrastructure can benefit from National Commission on Correctional Health Care standards as they are currently packaged, regardless of whether accreditation is the ultimate goal. PMID- 17403837 TI - Inflammatory markers and mediators in tracheal fluid of premature infants treated with inhaled nitric oxide. AB - OBJECTIVE: We compared serial measurements of inflammatory mediators and markers in infants treated with inhaled nitric oxide or placebo to assess the effects of inhaled nitric oxide therapy on lung inflammation during bronchopulmonary dysplasia. We investigated relationships between respiratory severity scores and airway concentrations of inflammatory markers/mediators. METHODS: As part of the Nitric Oxide (to Prevent) Chronic Lung Disease trial, a subset of 99 infants (52 placebo-treated infants and 47 inhaled nitric oxide-treated infants; well matched at baseline) had tracheal aspirate fluid collected at baseline, at 2 to 4 days, and then weekly while still intubated during study gas treatment (minimum of 24 days). Fluid was assessed for interleukin-1beta, interleukin-8, transforming growth factor-beta, N-acetylglucosaminidase, 8-epi-prostaglandin F2alpha, and hyaluronan. Results were normalized to total protein and secretory component of immunoglobulin A. RESULTS: At baseline, there was substantial variability of each measured substance and no correlation between tracheal aspirate fluid levels of any substance and respiratory severity scores. Inhaled nitric oxide administration did not result in any time-matched significant change for any of the analytes, compared with the placebo-treated group. There was no correlation between any of the measured markers/mediators and respiratory severity scores throughout the 24 days of study gas administration. In the posthoc analysis of data for inhaled nitric oxide-treated infants, there was a difference at baseline in 8-epi-prostaglandin F2alpha levels for infants who did (n = 21) and did not (n = 26) develop bronchopulmonary dysplasia at postmenstrual age of 36 weeks. CONCLUSIONS: Inhaled nitric oxide, as administered in this study, seemed to be safe. Its use was not associated with any increase in airway inflammatory substances. PMID- 17403838 TI - Do all infants with apparent life-threatening events need to be admitted? AB - OBJECTIVE: The goal was to identify criteria that would allow low-risk infants presenting with an apparent life-threatening event to be discharged safely from the emergency department. METHODS: We completed data forms prospectively on all previously healthy patients <12 months of age presenting to the emergency department of an urban tertiary care children's hospital with an apparent life threatening event over a 3-year period. These patients were then observed for subsequent events, significant interventions, or final diagnoses that would have mandated their admission (eg, sepsis). RESULTS: In our population of 59 infants, all 8 children who met the aforementioned outcome measures, thus requiring admission, either had experienced multiple apparent life-threatening events before presentation or were in their first month of life. In our study group, the high-risk criteria of age of <1 month [corrected] and multiple apparent life threatening events yielded a negative predictive value of 100% to identify the need for hospital admission. CONCLUSIONS: Our study suggests that >30-day-old infants who have experienced a single apparent life-threatening event may be discharged safely from the hospital, which would decrease admissions by 38%. PMID- 17403839 TI - Cost-effectiveness and potential impact of rotavirus vaccination in the United States. AB - OBJECTIVE: In February 2006, a safe, efficacious, orally administered pentavalent human-bovine reassortant rotavirus vaccine was licensed and recommended for routine immunization of all children in the United States. We assessed the health and economic impacts of a national rotavirus immunization program in the United States. METHODS: Monte Carlo cost-effectiveness analyses, from health care and societal perspectives, of vaccination of a hypothetical US birth cohort of 4,010,000 children monitored from birth to 59 months of age were performed. We compared the disease and economic burden of rotavirus infection in an unvaccinated cohort of children with one vaccinated at 2, 4, and 6 months with pentavalent human-bovine reassortant rotavirus vaccine. RESULTS: A routine rotavirus immunization program would prevent 13 deaths, 44,000 hospitalizations, 137,000 emergency department visits, 256,000 office visits, and 1,100,000 episodes requiring only home care for children <5 years of age in the United States. Assuming costs of administration of $10, the break-even price per dose of vaccine was $42 from the societal perspective and $12 from the health care perspective. From the societal perspective, at the manufacturer's price of $62.50 per dose, vaccination would cost $138 per case averted, $3024 per serious case averted, and $197,190 per life-year saved, at a total cost of $515 million to the health care system and $216 million to society. Key variables influencing the results were parental workdays lost, costs of hospitalization, emergency department visits, and child care. CONCLUSIONS: Despite a higher burden of serious rotavirus disease than estimated previously, routine rotavirus vaccination would unlikely be cost-saving in the United States at present. Nonetheless, rotavirus vaccination may still be considered a cost-effective intervention. PMID- 17403840 TI - Parental knowledge about antibiotic use: results of a cluster-randomized, multicommunity intervention. AB - OBJECTIVE: The goal was to determine the impact of a community-wide educational intervention on parental misconceptions likely contributing to pediatric antibiotic overprescribing. METHODS: We conducted a cluster-randomized trial of a 3-year, community-wide, educational intervention directed at parents of children < 6 years of age in 16 Massachusetts communities to improve parental antibiotic knowledge and attitudes and to decrease unnecessary prescribing. Parents in 8 intervention communities were mailed educational newsletters and exposed to educational materials during visits to local pediatric providers, pharmacies, and child care centers. We compared responses from mailed surveys in 2000 (before the intervention) and 2003 (after the intervention) for parents in intervention and control communities. Analyses were performed on the individual level, clustered according to community. RESULTS: There were 1106 (46%) and 2071 (40%) respondents to the 2000 and 2003 surveys, respectively. Between 2000 and 2003, the proportion of parents who answered > or = 7 of 10 knowledge questions correctly increased significantly in both intervention (from 52% to 64%) and control (from 54% to 61%) communities. We did not detect a significant intervention impact on knowledge regarding appropriate antibiotic use in the population overall. In a subanalysis, we did observe a significant intervention effect among parents of Medicaid-insured children, who began with lower baseline knowledge scores. CONCLUSIONS: Although knowledge regarding appropriate use of antibiotics is improving without additional targeted intervention among more socially advantaged populations, parents of Medicaid-insured children may benefit from educational interventions to promote judicious antibiotic use. These findings may have implications for other health education campaigns. PMID- 17403841 TI - Reduction of frequent otitis media and pressure-equalizing tube insertions in children after introduction of pneumococcal conjugate vaccine. AB - OBJECTIVE: Streptococcus pneumoniae is an important cause of otitis media in children. In this study we estimated the effect of routine childhood immunization with heptavalent pneumococcal conjugate vaccine on frequent otitis media (3 episodes in 6 months or 4 episodes in 1 year) and pressure-equalizing tube insertions. PATIENTS AND METHODS: The study population included all children who were enrolled at birth in TennCare or selected upstate New York commercial insurance plans as of July 1998 and continuously followed until 5 years old, loss of health plan enrollment, study outcome, or end of the study. We compared the risk of developing frequent otitis media or having pressure-equalizing tube insertion for 4 birth cohorts (1998-1999, 1999-2000, 2000-2001, and 2001-2002) by using Cox regression analysis. We used data from the National Immunization Survey to estimate the heptavalent pneumococcal conjugate vaccine uptake for children in these 4 birth cohorts in Tennessee and New York. RESULTS: The proportion of children in Tennessee and New York who received at least 3 doses of heptavalent pneumococcal conjugate vaccine by 2 years of age increased from < or = 1% for the 1998-1999 birth cohort to approximately 75% for the 2000-2001 birth cohort. By age 2 years, 29% of Tennessee and New York children born in 2000-2001 had developed frequent otitis media, and 6% of each of these birth cohorts had pressure-equalizing tubes inserted. Comparing the 2000-2001 birth cohort to the 1998-1999 birth cohort, frequent otitis media declined by 17% and 28%, and pressure-equalizing tube insertions declined by 16% and 23% for Tennessee and New York children, respectively. For the 2000-2001 to the 2001-2002 birth cohort, frequent otitis media and pressure-equalizing tubes remained stable in New York but increased in Tennessee. CONCLUSIONS: After heptavalent pneumococcal conjugate vaccine introduction, children were less likely to develop frequent otitis media or have pressure-equalizing tube insertions. PMID- 17403842 TI - Outcome at 2 years of age of infants from the DART study: a multicenter, international, randomized, controlled trial of low-dose dexamethasone. AB - OBJECTIVE: Low-dose dexamethasone facilitates extubation in chronically ventilator-dependent infants with no obvious short-term complications. The objective of this study was to determine the long-term effects of low-dose dexamethasone. METHODS: Very preterm (<28 weeks' gestation) or extremely low birth weight (birth weight <1000 g) infants who were ventilator dependent after the first week of life for whom clinicians considered corticosteroids were indicated were eligible. After informed consent, infants were randomly assigned to masked dexamethasone (0.89 mg/kg over 10 days) or saline placebo. Survivors were assessed at 2 years' corrected age by staff blinded to treatment group allocation to determine neurosensory outcome, growth, and health. RESULTS: The trial was abandoned well short of its target sample size because of recruitment difficulties. Seventy infants were recruited from 11 centers, 35 in each group: 59 survived to 2 years of age, and 58 (98%) were assessed at follow-up, but data for cerebral palsy were available for only 56 survivors. There was little evidence for a difference in the major end point, the rate of the combined outcome of death, or major disability at 2 years of age (dexamethasone group: 46%; controls: 43%). Rates of mortality before follow-up (11% vs 20%), major disability (41% vs 31%), cerebral palsy (14% vs 22%), or of the combined outcomes of death or cerebral palsy (23% vs 37%) were not substantially different between the groups. There were no obvious effects of low-dose dexamethasone on growth or readmissions to hospital after discharge. CONCLUSIONS: Although this trial was not able to provide definitive evidence on the long-term effects of low-dose dexamethasone after the first week of life in chronically ventilator-dependent infants, our data indicate no strong association with long-term morbidity. PMID- 17403843 TI - Long-term outcome and clinical spectrum of 73 pediatric patients with mitochondrial diseases. AB - OBJECTIVES: We sought to determine the clinical spectrum, survival, and long-term functional outcome of a cohort of pediatric patients with mitochondrial diseases and to identify prognostic factors. METHODS: Medical charts were reviewed for 73 children diagnosed between 1985 and 2005. The functional status of living patients was assessed prospectively by using the standardized Functional Independence Measure scales. RESULTS: Patients fell into 7 phenotypic categories: neonatal-onset lactic acidosis (10%), Leigh syndrome (18%), nonspecific encephalopathy (32%), mitochondrial (encephalo)myopathy (19%), intermittent neurologic (5%), visceral (11%), and Leber hereditary optic neuropathy (5%). Age at first symptoms ranged from prenatal to 16 years (median: 7 months). Neurologic symptoms were the most common (90%). Visceral involvement was observed in 29% of the patients. A biochemical or molecular diagnosis was identified for 81% of the patients as follows: deficiency of complex IV (27%), of pyruvate dehydrogenase or complex I (25% each), of multiple complexes (13%), and of pyruvate carboxylase (5%) or complexes II+III (5%). A mitochondrial DNA mutation was found in 20% of patients. At present, 46% of patients have died (median age: 13 months), 80% of whom were <3 years of age. Multivariate analysis showed that age at first symptoms was a major independent predictor of mortality: patients with first symptoms before 6 months had a highly increased risk of mortality. Cardiac or visceral involvement and neurologic crises were not independent prognostic factors. Living patients showed a wide range of independence levels that correlated positively with age at first symptoms. Among patients aged >5 years (n = 32), 62% had Functional Independence Measure quotients of >0.75. CONCLUSIONS: Mitochondrial diseases in children span a wide range of symptoms and severities. Age at first symptoms is the strongest predictor mortality. Despite a high mortality rate in the cohort, 62% of patients aged >5 years have only mild impairment or normal functional outcome. PMID- 17403844 TI - Association between passive smoking and infection with Mycobacterium tuberculosis in children. AB - OBJECTIVE: Tuberculosis and smoking are both significant public health problems. The association between passive smoking and Mycobacterium tuberculosis infection is not well documented. The objective of this study was to examine the influence of passive smoking on M. tuberculosis infection in children. METHODS: A community survey was conducted in 15% of addresses in 2 adjacent low-income suburbs in Cape Town, South Africa. All children (< 15 years of age) and their adult household members residing at these addresses were included in the study. Children underwent tuberculin skin testing. An induration of > or = 10 mm was considered to define M. tuberculosis infection. Passive smoking was defined as living in the household with at least 1 adult who smoked for at least 1 year. Random-effects logistic regression analysis was performed, and odds ratios were adjusted for age, presence of a patient with tuberculosis in the household, average household income, and clustering at the household level. RESULTS: Of 1344 children, 432 (32%) had a positive tuberculin skin test. Passive smoking was significantly associated with M. tuberculosis infection in the unadjusted analyses but not in the adjusted analyses. In the 172 households with a patient with tuberculosis, passive smoking was significantly associated with a positive tuberculin skin test but not in the 492 households without a patient with tuberculosis. CONCLUSIONS: Passive smoking is associated with M. tuberculosis infection in children living in a household with a patient with tuberculosis. More studies are needed to confirm this observation, but the possible association is a cause of great concern, considering the high prevalence of smoking and tuberculosis in most developing countries. PMID- 17403845 TI - Incidence, complications, and risk factors for prolonged stay in children hospitalized with community-acquired influenza. AB - OBJECTIVES: Few studies have examined the characteristics and clinical course of children hospitalized with laboratory-confirmed influenza. We sought to (1) estimate the age-specific incidence of influenza-related hospitalizations, (2) describe the characteristics and clinical course of children hospitalized with influenza, and (3) identify risk factors for prolonged hospitalization. PATIENTS AND METHODS: Children < or = 21 years of age hospitalized with community-acquired laboratory-confirmed influenza at a large urban children's hospital were identified through review of laboratory records and administrative data sources. A neighborhood cohort embedded within our study population was used to estimate the incidence of community-acquired laboratory-confirmed influenza hospitalizations among children < 18 years old. Risk factors for prolonged hospitalization (> 6 days) were determined by using logistic regression. RESULTS: We identified 745 children hospitalized with community-acquired laboratory confirmed influenza during the 4-year study period. In this urban cohort, the incidence of community-acquired laboratory-confirmed influenza hospitalization was 7 per 10,000 child-years of observation. The median age was 1.8 years; 25% were infants < 6 months old, and 77% were children < 5 years old. Many children (49%) had a medical condition associated with an increased risk of influenza related complications. The incidence of influenza-related complications was higher among children with a preexisting high-risk condition than for previously healthy children (29% vs 21%). However, only cardiac and neurologic/neuromuscular diseases were found to be independent risk factors for prolonged hospitalization. CONCLUSIONS: Influenza is a common cause of hospitalization among both healthy and chronically ill children. Children with cardiac or neurologic/neuromuscular disease are at increased risk of prolonged hospitalization; therefore, children with these conditions and their contacts should be a high priority to receive vaccine. The impact on pediatric hospitalization of the new recommendation to vaccinate all children 6 months to < 5 years old should be assessed. PMID- 17403846 TI - Previous head injury is a risk factor for subsequent head injury in children: a longitudinal cohort study. AB - OBJECTIVE: The objective of this study was to determine whether children who sought care for a head injury were at greater risk of having a subsequent head injury within the following 6 and 12 months compared with children who sought care for an injury other than to the head. DESIGN/SETTING: This was a longitudinal cohort study conducted in the emergency departments of 2 Montreal (Quebec, Canada) pediatric hospitals. PARTICIPANTS: The parents of 11,867 injured children aged 1 to 18 years were interviewed by telephone at 6 (n = 10,315) and 12 (n = 9486) months after their child's injury to ascertain outcome (ie, subsequent head injury) and to provide information on potential risk factors (age, gender, chronic medical condition, activity level, and socioeconomic status). MAIN OUTCOME MEASURE: The outcome of interest was a head injury requiring medical attention within the following year ascertained by parental recall or physician claims data. RESULTS: A total of 245 and 386 previously head injured children sustained a subsequent head injury within 6 and 12 months, respectively. Children who sought care for an initial head injury (n = 3599) were at higher risk of having a subsequent head injury within 6 months than children who sought care for an injury not to the head (n = 6716). The adjusted odds ratio suggested weak confounding by age, gender, and history of previous head injury. Results were consistent on the basis of physician claims data and 12-month follow up interview data. CONCLUSIONS: These results provide evidence that having a head injury increases a child's risk of having a subsequent head injury. Although age, gender, and history of previous head injury confound the relationship, the effect remains substantial. PMID- 17403847 TI - Quantification of deep gray matter in preterm infants at term-equivalent age using manual volumetry of 3-tesla magnetic resonance images. AB - OBJECTIVE: Nonhypothesis-based MRI-analysis techniques including deformation based morphometry and automated tissue segmentation have suggested that preterm infants at term-equivalent age have reduced tissue volume in the basal ganglia and thalami, which is most apparent among infants with supratentorial lesions. The aim of our study was to test this hypothesis by direct measurement of thalamic and lentiform nuclei volumes in preterm infants at term-equivalent age and term-born controls using manual volumetry. DESIGN/METHODS: Forty preterm infants at term-equivalent age (median gestational age: 29.5 weeks; median birth weight: 1.3 kg) and 8 term-born controls were examined using a 3-T Philips (Best, Netherlands) system. T1-weighted volume images and T2-weighted fast-spin echo pseudovolumes were acquired. There was no significant difference in postmenstrual age at image acquisition between the 2 groups. ImageJ 1.34 (National Institutes of Health, Bethesda, MD) was used for manual segmentations. RESULTS: The median thalamic and lentiform nuclei volumes for preterm infants at term-equivalent age were 13.6 and 3.07 cm3, respectively, significantly smaller than term-control volumes of 16.3 and 5.6 cm3, respectively. Ten preterm infants at term-equivalent age had supratentorial lesions (intraventricular hemorrhage, periventricular leukomalacia, or hemorrhagic parenchymal infarction), and the median thalamic and lentiform volumes for this group were 10.4 and 1.7 cm3, respectively. When this group was excluded, the remaining infants who had mild or moderate diffuse excessive high signal intensity in the white matter on T2-weighted images had a smaller, yet significant, volume reduction compared with controls. Tissue volumes were not related to weight and gestational age at birth. CONCLUSIONS: Manual volumetry confirms that preterm infants at term-equivalent age have reduced thalamic and lentiform volumes compared with controls. This was most marked among infants with supratentorial lesions but was also seen among those with nonfocal white matter abnormalities. PMID- 17403848 TI - Paracetamol (acetaminophen) penetrates readily into the cerebrospinal fluid of children after intravenous administration. AB - INTRODUCTION: The main action of paracetamol (acetaminophen) is presumed to be in the central nervous system. The central nervous system penetration of paracetamol has been described in children with intracranial pathologies but not in children with an intact blood-brain barrier. OBJECTIVE: We investigated the cerebrospinal fluid penetration of paracetamol in 32 healthy children, aged 3 months to 12 years, who were undergoing surgery in the lower body using spinal anesthesia. MATERIALS AND METHODS: In this open-label prospective study, children were given a single intravenous injection of paracetamol (15 mg/kg). Cerebrospinal fluid and venous blood samples were obtained between 5 minutes and 5 hours after injection. Paracetamol concentrations were determined from the cerebrospinal fluid and plasma by using a fluorescence polarization immunoassay. RESULTS: Paracetamol was detected in cerebrospinal fluid from the earliest sample at 5 minutes, although in this sample paracetamol concentration was below the limit of quantification of 1.0 mg/L. Subsequent paracetamol concentrations in cerebrospinal fluid ranged between 1.3 and 18 mg/L (median: 7.2 mg/L), plasma concentrations ranged between 2.4 and 33 mg/L, and cerebrospinal fluid/plasma ratios ranged between 0.06 and 2.0. The highest CSF paracetamol concentration was detected at 57 minutes. CONCLUSIONS: Paracetamol permeates readily into the cerebrospinal fluid of children. This fast and extensive transfer enables the rapid central analgesic and antipyretic action of intravenous paracetamol. PMID- 17403849 TI - Antifungal therapy in children with invasive fungal infections: a systematic review. AB - Invasive fungal infections are associated with significant morbidity and mortality. Differences between children and adults are reported, yet few trials of antifungal agents have been performed in pediatric populations. We performed a systematic review of the literature to guide appropriate pediatric treatment recommendations. From available trials that compared antifungal agents in either prolonged febrile neutropenia or invasive candidal or Aspergillus infection, no clear difference in treatment efficacy was demonstrated, although few trials were adequately powered. Differing antifungal pharmacokinetics between children and adults were demonstrated, requiring dose modification. Significant differences in toxicity, particularly nephrotoxicity, were identified between classes of antifungal agents. Therapy needs to be guided by the pathogen or suspected pathogens, the degree of immunosuppression, comorbidities (particularly renal dysfunction), concurrent nephrotoxins, and the expected length of therapy. PMID- 17403850 TI - Creation and development of the public service orphan drug Human Botulism Immune Globulin. AB - The public service orphan drug Human Botulism Immune Globulin for the treatment of infant botulism would not have come into existence without the federal Orphan Drug Act and the funding mechanism that it provided to conduct pivotal clinical trials. Nonetheless, creating, developing, and achieving licensure of Human Botulism Immune Globulin took approximately 15 years and approximately $10.6 million (2005 dollars) to accomplish. Use of Human Botulism Immune Globulin to treat patients with infant botulism has resulted thus far in more than 30 years of avoided hospital stay and more than $50 million (2005 dollars) of avoided hospital costs. To provide a possible paradigm for others, the circumstances that enabled a state public health department to create, test, license, and distribute an orphan drug are described here. PMID- 17403851 TI - Executive summary of the workshop on oxygen in neonatal therapies: controversies and opportunities for research. AB - One of the most complex areas in perinatal/neonatal medicine is the use of oxygen in neonatal therapies. To address the knowledge gaps that preclude optimal, evidence-based care in this critical field of perinatal medicine, the National Institute of Child Health and Human Development organized a workshop, Oxygen in Neonatal Therapies: Controversies and Opportunities for Research, in August 2005. The information presented at the workshop included basic and translational oxygen research; a review of completed, ongoing, and planned clinical trials; oxygen administration for neonatal resuscitation; and a review of the collaborative home infant monitoring evaluation study. This article provides a summary of the discussions, focusing on major knowledge gaps, with prioritized suggestions for studies in this area. PMID- 17403852 TI - The intimidation of British pediatricians. AB - British pediatricians who diagnose and treat child abuse cases have come under attack by the British press and by parents who have been investigated for possible abuse. Now the General Medical Council also is intimidating these pediatricians. The General Medical Council is the licensing authority for physicians in the United Kingdom. This has resulted in fewer pediatricians being willing to care for abused children or to testify in child abuse cases. In the United States, the recent recognition of the pediatric subspecialty of child abuse pediatrics should help set standards for child abuse medical evaluation and testimony. PMID- 17403853 TI - United Kingdom General Medical Council fails child protection. AB - To protect children, pediatricians must be willing to raise the possibility of abuse and not be intimidated by the consequences. We consider that the United Kingdom General Medical Council does not understand child protection matters and has no system for dealing adequately with complaints submitted by parents who claim false allegations of abuse. The actions of the General Medical Council in the recent cases of Drs Roy Meadow and David Southall conflict with current child protection laws and guidance for professionals. By deterring doctors from raising concerns about a child's safety and giving opinions on child deaths, the General Medical Council may be increasing the risk of serious child abuse. Although the rate of registrations by child protection authorities decreased by 28% between 1995 and 2005 (ie, there are fewer multiagency child protection plans), the number of criminal convictions for cruelty to or neglect of a child increased by 247% between 1998 and 2005. It is unacceptable that to date the General Medical Council has refused training in child protection offered by the Royal College of Paediatrics and Child Health. PMID- 17403854 TI - Is there a strategy for preventing bronchopulmonary dysplasia? Absence of evidence is not evidence of absence. PMID- 17403855 TI - Male circumcision for prevention of HIV and other sexually transmitted diseases. PMID- 17403856 TI - State Children's Health Insurance Program reauthorization: will it get us closer to universal coverage for America's children? PMID- 17403857 TI - Clinical mimics of infant botulism. AB - Since 1992, Human Botulism Immune Globulin has been provided by the California Department of Health Services to infants with probable infant botulism, the intestinal toxemia form of human botulism. Human Botulism Immune Globulin became available in California in 1992-1997 within a randomized, controlled, double blinded, pivotal clinical trial and subsequently became available nationwide in 1998-2003 in an open-label study until its licensure in October 2003 as BabyBIG. Thereafter, Human Botulism Immune Globulin remained available nationwide as an approved orphan-drug product. To achieve prompt neutralization of circulating botulinum toxin, the decision to treat with Human Botulism Immune Globulin has been based on clinical criteria that include a consistent history and physical findings of bulbar palsies, hypotonia, and weakness. After licensure, the charts of patients who did not have laboratory-confirmed infant botulism were reviewed to identify their actual diagnoses. The approximately 5% of 681 patients treated with Human Botulism Immune Globulin who did not have infant botulism fell into 5 categories: spinal muscular atrophy, metabolic disorders, other infectious diseases, miscellaneous, and probable infant botulism lacking laboratory confirmation. PMID- 17403858 TI - Family-centered bedside rounds: a new approach to patient care and teaching. AB - The importance of patient-centered care and the role of families in decision making are becoming more recognized. Starting with a single acute care unit, a multidisciplinary improvement team at Cincinnati Children's Hospital developed and implemented a new process that allows families to decide if they want to be part of attending-physician rounds. Family involvement seems to improve communication, shares decision-making, and offers new learning for residents and students. Despite initial concerns of staff members, family-centered rounds has been widely accepted and spread throughout the institution. Here we report our experiences as a potential model to improve family-centered care and teaching. PMID- 17403859 TI - Enhancing the diversity of the pediatrician workforce. AB - This policy statement describes the key issues related to diversity within the pediatrician and health care workforce to identify barriers to enhancing diversity and offer policy recommendations to overcome these barriers in the future. The statement addresses topics such as health disparities, affirmative action, recent policy developments and reports on workforce diversity, and research on patient and provider diversity. It also broadens the discussion of diversity beyond the traditional realms of race and ethnicity to include cultural attributes that may have an effect on the quality of health care. Although workforce diversity is related to the provision of culturally effective pediatric care, it is a discrete issue that merits separate discussion and policy formulation. At the heart of this policy-driven action are multiorganizational and multispecialty collaborations designed to address substantive educational, financial, organizational, and other barriers to improved workforce diversity. PMID- 17403860 TI - Increasing antiretroviral drug access for children with HIV infection. AB - Although there have been great gains in the prevention of pediatric HIV infection and provision of antiretroviral therapy for children with HIV infection in resource-rich countries, many barriers remain to scaling up HIV prevention and treatment for children in resource-limited areas of the world. Appropriate testing technologies need to be made more widely available to identify HIV infection in infants. Training of practitioners in the skills required to care for children with HIV infection is required to increase the number of children receiving antiretroviral therapy. Lack of availability of appropriate antiretroviral drug formulations that are easily usable and inexpensive is a major impediment to optimal care for children with HIV. The time and energy spent trying to develop liquid antiretroviral formulations might be better used in the manufacture of smaller pill sizes or crushable tablets, which are easier to dispense, transport, store, and administer to children. PMID- 17403861 TI - Prevention of influenza: recommendations for influenza immunization of children, 2006-2007. AB - The purpose of this statement is to update recommendations for routine use of influenza vaccine in children for the 2006-2007 influenza season. The American Academy of Pediatrics recommends annual influenza immunization for (1) children with high-risk conditions who are 6 months and older; (2) healthy children 6 through 59 months of age; (3) household contacts and out-of-home caregivers of children with high-risk conditions and all healthy children younger than 5 years; and (4) health care professionals. Other children, adolescents, and adults can be immunized to decrease the impact of influenza as indicated in the Red Book: 2006 Report of the Committee on Infectious Diseases. PMID- 17403862 TI - Antiviral therapy and prophylaxis for influenza in children. AB - Antiviral agents are available that are safe and effective for the treatment and prophylaxis of influenza virus infections in children. The neuraminidase inhibitors (oseltamivir [Tamiflu] and zanamivir [Relenza]) are preferred agents because of current widespread resistance to the adamantanes (amantadine [Symmetrel] and rimantadine [Flumadine]). Therapy should be provided to children with influenza infection who are at high risk of severe infection and to children with moderate-to-severe influenza infection who may benefit from a decrease in the duration of symptoms. Prophylaxis should be provided (1) to high-risk children who have not yet received immunization and during the 2 weeks after immunization, (2) to unimmunized family members and health care professionals with close contact with high-risk unimmunized children or infants who are younger than 6 months, and (3) for control of influenza outbreaks in unimmunized staff and children in an institutional setting. Testing of current H5N1 avian influenza virus isolates, the potential agents of pandemic influenza, suggests susceptibility to oseltamivir and zanamivir. Because no prospective data exist on the efficacy of these agents in humans for H5N1 strains, the dosage and duration of therapy in adults and children may differ from those documented to be effective for epidemic influenza strains. PMID- 17403863 TI - Issues with the new developmental screening and surveillance policy statement. PMID- 17403864 TI - Evidence-based medicine practice by hospitalists and community pediatricians. PMID- 17403865 TI - High-dose systemic corticosteroids may be effective early in the course of bronchiolitis. PMID- 17403866 TI - Involvement of caspase-9 in the inhibition of necrosis of RAW 264 cells infected with Mycobacterium tuberculosis. AB - In order to know how caspases contribute to the intracellular fate of Mycobacterium tuberculosis and host cell death in the infected macrophages, we examined the effect of benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethane (z-VAD fmk), a broad-spectrum caspase inhibitor, on the growth of M. tuberculosis H37Rv in RAW 264 cells. In the cells treated with z-VAD-fmk, activation of caspase-8, caspase-3/7, and caspase-9 was clearly suppressed, and DNA fragmentation of the infected cells was also reduced. Under this experimental condition, it was found that the treatment markedly inhibited bacterial growth inside macrophages. The infected cells appeared to undergo cell death of the necrosis type in the presence of z-VAD-fmk. We further found that z-VAD-fmk treatment resulted in the generation of intracellular reactive oxygen species (ROS) in the infected cells. By addition of a scavenger of ROS, the host cell necrosis was inhibited and the intracellular growth of H37Rv was significantly restored. Among inhibitors specific for each caspase, only the caspase-9-specific inhibitor enhanced the generation of ROS and induced necrosis of the infected cells. Furthermore, we found that severe necrosis was induced by infection with H37Rv but not H37Ra in the presence of z-VAD-fmk. Caspase-9 activation was also detected in H37Rv infected cells, but H37Ra never induced such caspase-9 activation. These results indicated that caspase-9, which was activated by infection with virulent M. tuberculosis, contributed to the inhibition of necrosis of the infected host cells, presumably through suppression of intracellular ROS generation. PMID- 17403867 TI - Reduced pathology following infection with transgenic Leishmania major expressing murine CD40 ligand. AB - Leishmanization is the inoculation of live Leishmania into the host to vaccinate against subsequent infections. This approach has been largely discontinued due to safety concerns. We have previously shown that combining CD40 ligand (CD40L) with Leishmania antigen preferentially induces a type 1 immune response and provides some protection to vaccinated mice (G. Chen, P. A. Darrah, and D. M. Mosser, Infect. Immun. 69:3255-3263, 2001). In the present study, we developed transgenic L. major organisms which express and secrete the extracellular portion of CD40L (L. major CD40LE). We hypothesized that these organisms would be less virulent but more immunogenic than wild-type organisms and therefore be more effective at leishmanization. Transgenic parasites expressing CD40L mRNA and protein were developed. BALB/c mice infected with these parasites developed significantly smaller lesions containing fewer parasites than animals infected with wild-type organisms. Infection of resistant C57BL/6 mice with low doses of transgenic parasites induced a significant amount of protection against subsequent high-dose infection with wild-type organisms. These results demonstrate that transgenic organisms expressing CD40L are less virulent than wild-type organisms while retaining full immunogenicity. PMID- 17403868 TI - Moraxella catarrhalis outer membrane protein CD elicits antibodies that inhibit CD binding to human mucin and enhance pulmonary clearance of M. catarrhalis in a mouse model. AB - The outer membrane protein CD of Moraxella catarrhalis is considered to be a potential vaccine antigen against Moraxella infection. We purified the native CD from isolate O35E, administered it to mice, and detected considerable titers of anti-CD antibodies. Anti-CD sera were cross-reactive towards six different M. catarrhalis isolates and promoted bacterial clearance of O35E in a pulmonary challenge model. To circumvent the difficulty of generating large quantities of CD from M. catarrhalis for vaccine use, the CD gene from O35E was cloned into Escherichia coli, and the recombinant CD, expressed without a signal sequence or fusion tags, represented approximately 70% of the total E. coli proteins. The recombinant CD formed inclusion bodies that were solubilized with 6 M urea and then purified by ion-exchange chromatography, a procedure that produced soluble CD of high purity and yield. Mice immunized with the purified recombinant CD had significant titers of anti-CD antibodies that were cross-reactive towards 24 different M. catarrhalis isolates. Upon challenge, these mice showed enhanced bacterial clearance of both O35E and a heterologous M. catarrhalis isolate, TTA24. In an in vitro assay, antisera to either the native or the recombinant CD inhibited the binding activity of CD to human tracheobronchial mucin in a serum concentration-dependent manner, and the extent of inhibition appeared to correlate with the corresponding anti-CD antibody titer and whole-cell enzyme linked immunosorbent assay titer. Our results demonstrate that the recombinant CD is a promising vaccine candidate for preventing Moraxella infection. PMID- 17403869 TI - The type IV pilin of Burkholderia mallei is highly immunogenic but fails to protect against lethal aerosol challenge in a murine model. AB - Burkholderia mallei is the cause of glanders and a proven biological weapon. We identified and purified the type IV pilin protein of this organism to study its potential as a subunit vaccine. We found that purified pilin was highly immunogenic. Furthermore, mice infected via sublethal aerosol challenge developed significant increases in titers of antibody against the pilin, suggesting that it is expressed in vivo. Nevertheless, we found no evidence that high-titer antipilin antisera provided passive protection against a sublethal or lethal aerosol challenge and no evidence of protection afforded by active immunization with purified pilin. These results contrast with the utility of type IV pilin subunit vaccines against other infectious diseases and highlight the need for further efforts to identify protective responses against this pathogen. PMID- 17403870 TI - Filamentous influenza A virus infection predisposes mice to fatal septicemia following superinfection with Streptococcus pneumoniae serotype 3. AB - Previous studies have demonstrated that animals exposed to Streptococcus pneumoniae while recovering from influenza A virus infection exhibit exacerbated disease symptoms. However, many of the current animal models exploring dual viral and bacterial synergistic exacerbations of respiratory disease have utilized mouse-adapted influenza virus and strains of Streptococcus pneumoniae that in themselves are highly lethal to mice. Here we describe a mouse model of bacterial superinfection in which a mild, self-limiting influenza virus infection is followed by mild, self-limiting superinfection with S. pneumoniae serotype 3. S. pneumoniae superinfection results in rapid dissemination of the bacterium from the respiratory tract and systemic spread to all major organs of the mice, resulting in fatal septicemia. This phenomenon in mice was observed in superinfected animals undergoing an active viral infection as well as in mice that had completely cleared the virus 7 to 8 days prior to superinfection. Neutrophils were the predominant cellular inflammatory infiltrate in the lungs of superinfected mice compared to singly infected animals. Among other cytokines and chemokines, the neutrophil activator granulocyte colony-stimulating factor (G CSF) was found to be significantly overexpressed in the spleens, lungs, and brains of superinfected animals. High G-CSF protein levels were observed in sera and lung lavage fluid from superinfected animals, suggesting that G-CSF is a major contributor to synergistic exacerbation of disease leading to fatal septicemia. PMID- 17403871 TI - Systemic CD8 T-cell memory response to a Salmonella pathogenicity island 2 effector is restricted to Salmonella enterica encountered in the gastrointestinal mucosa. AB - To better understand the evolution of a systemic memory response to a mucosal pathogen, we monitored antigen-specific OT1 CD8 T-cell responses to a fusion of the SspH2 protein and the peptide SIINFEKL stably expressed from the chromosome of Salmonella enterica and loaded into the class I pathway of antigen presentation of professional phagocytes through the Salmonella pathogenicity island 2 type III secretion system (TTSS). This strategy has revealed that effector memory CD8 T cells with low levels of CD62L expression (CD62L(low)) are maintained in systemic sites months after vaccination in response to low-grade infections with Salmonella. However, the CD8 T-cell pool eventually declines. Low numbers of central memory cells surviving after prolonged resting from an antigen encounter can nevertheless reconstitute the systemic effector memory pool in a route-specific recall response to cognate antigens encountered in the gut. Accordingly, populations of CD62L(high) interleukin-7 receptor-positive progenitor central memory cells grafted into naive mice expand in response to orally administered Salmonella expressing the chromosomal translational fusion of sspH2 and the sequence encoding the SIINFEKL peptide but fail to proliferate following systemic stimulation. Moreover, populations of systemic memory CD8 T cells restricted to Salmonella in oral vaccines selectively expand in response to cognate antigens presented by cells isolated from mesenteric lymph nodes (MLN). Together, these findings have revealed the imprinting of systemic CD8 central memory T-cell recall responses against enteropathogens by MLN. MLN restriction represents a novel mechanism by which systemic CD8 T-cell immunity is confined to periods of high risk for extraintestinal dissemination. PMID- 17403872 TI - RNA interference-mediated silencing of the YPS3 gene of Histoplasma capsulatum reveals virulence defects. AB - The YPS3 gene of Histoplasma capsulatum encodes a protein that is both surface localized in the cell wall of H. capsulatum and released into the culture medium. This protein is produced only during the pathogenic yeast phase of infection and is also expressed differentially in H. capsulatum strains of different virulence levels. In this study, we silenced the YPS3 transcript by using an interfering RNA strategy and examined the silenced mutants for phenotypic differences in vitro and during infection. The mutants showed no growth defect during in vitro culture in a defined medium at 37 degrees C and appeared to have normal virulence in a RAW 264.7 murine macrophage-like cell line. In a C57BL/6 mouse model of infection, however, the mutants caused significantly decreased fungal burdens, particularly in the peripheral phagocyte-rich tissues of livers and spleens. This defect in organ colonization was evident within 3 days of infection; however, it appeared to be exacerbated at later time points. PMID- 17403873 TI - Interleukin-23 (IL-23)-IL-17 cytokine axis in murine Pneumocystis carinii infection. AB - Host defense mechanisms against Pneumocystis carinii are not fully understood. Previous work in the murine model has shown that host defense against infection is critically dependent upon host CD4(+) T cells. The recently described Th17 immune response is predominantly a function of effector CD4(+) T cells stimulated by interleukin-23 (IL-23), but whether these cells are required for defense against P. carinii infection is unknown. We tested the hypothesis that P. carinii stimulates the early release of IL-23, leading to increases in IL-17 production and lung effector CD4(+) T-cell population that mediate clearance of infection. In vitro, stimulation of alveolar macrophages with P. carinii induced IL-23, and IL-23p19 mRNA was expressed in lungs of mice infected with this pathogen. To address the role of IL-23 in resistance to P. carinii, IL-23p19-/- and wild-type control C57BL/6 mice were infected and their fungal burdens and cytokine/chemokine responses were compared. IL-23p19-/- mice displayed transient but impaired clearance of infection, which was most apparent 2 weeks after inoculation. In confirmatory studies, the administration of either anti-IL-23p19 or anti-IL-17 neutralizing antibody to wild-type mice infected with P. carinii also caused increases in fungal burdens. IL-17 and the lymphocyte chemokines IP 10, MIG, MIP-1alpha, MIP-1beta, and RANTES were decreased in the lungs of infected IL-23p19-/- mice in comparison to their levels in the lungs of wild-type mice. In IL-23p19-/- mice infected with P. carinii, there were fewer effector CD4(+) T cells in the lung tissue. Collectively, these studies indicate that the IL-23-IL-17 axis participates in host defense against P. carinii. PMID- 17403874 TI - CD14 mediates cross talk between mononuclear cells and fibroblasts for upregulation of matrix metalloproteinase 9 by Borrelia burgdorferi. AB - Lyme disease is an infection caused by a tick-borne spirochete, Borrelia burgdorferi. Matrix metalloproteinase 9 (MMP-9) was selectively upregulated in the erythema migrans skin lesions of patients with acute Lyme disease. In this study, the mechanism of upregulation of MMP-9 was investigated in vitro and in vivo. The concentrations of MMP-9 and soluble CD14 were markedly elevated in serum from patients with acute Lyme disease and were also upregulated in U937 cells by B. burgdorferi in a time- and concentration-dependent manner. MMP-9 mRNA was expressed at baseline in fibroblasts in the presence or absence of B. burgdorferi. However, when fibroblasts were incubated with supernatants from U937 cells with B. burgdorferi or recombinant CD14, the expression of MMP-9 was significantly increased. This effect was completely abolished by the anti-CD14 antibody. These data suggest that the upregulation of MMP-9 by B. burgdorferi involves the CD14 pathway in infiltrating inflammatory cells. Fibroblasts could be recruited to amplify local production of MMP-9 by acquiring CD14 from macrophages. PMID- 17403875 TI - Differential interaction of dendritic cells with Rickettsia conorii: impact on host susceptibility to murine spotted fever rickettsiosis. AB - Spotted fever group rickettsioses are emerging and reemerging infectious diseases, some of which are life-threatening. In order to understand how dendritic cells (DCs) contribute to the host resistance or susceptibility to rickettsial diseases, we first characterized the in vitro interaction of rickettsiae with bone marrow-derived DCs (BMDCs) from resistant C57BL/6 (B6) and susceptible C3H/HeN (C3H) mice. In contrast to the exclusively cytosolic localization within endothelial cells, rickettsiae efficiently entered and localized in both phagosomes and cytosol of BMDCs from both mouse strains. Rickettsia conorii-infected BMDCs from resistant mice harbored higher bacterial loads compared to C3H mice. R. conorii infection induced maturation of BMDCs from both mouse strains as judged by upregulated expression of classical major histocompatibility complex (MHC) and costimulatory molecules. Compared to C3H counterparts, B6 BMDCs exhibited higher expression levels of MHC class II and higher interleukin-12 (IL-12) p40 production upon rickettsial infection and were more potent in priming naive CD4(+) T cells to produce gamma interferon. In vitro DC infection and T-cell priming studies suggested a delayed CD4(+) T-cell activation and suppressed Th1/Th2 cell development in C3H mice. The suppressive CD4(+) T-cell responses seen in C3H mice were associated with a high frequency of Foxp3(+) T regulatory cells promoted by syngeneic R. conorii-infected BMDCs in the presence of IL-2. These data suggest that rickettsiae can target DCs to stimulate a protective type 1 response in resistant hosts but suppressive adaptive immunity in susceptible hosts. PMID- 17403876 TI - CD4+ lymphocytes and gamma interferon predominate in local immune responses in early experimental syphilis. AB - The clearance of Treponema pallidum subsp. pallidum from early syphilis lesions involves infiltration of a large number of mononuclear cells and is characteristic of a cell-mediated immune response. In the present study, we sought to determine the relative abundance of different T-lymphocyte populations and Th1/Th2-associated cytokines present in testicular lesions following experimental infection with the Chicago strain of T. pallidum. Using flow cytometry, we examined the proportion of CD4(+) and CD8(+) T cells present throughout the progression and resolution of primary syphilis in the rabbit model. We related these findings to the results of real-time reverse transcription-PCR quantification of treponemal and cytokine mRNA levels. Treponemal mRNA levels reached peak values on day 18 postinfection, coincident with an initial peak in the level of T cells, which were primarily CD4(+) T cells. T-cell levels increased again during resolution of orchitis, and there was an increased proportion of CD8(+) T cells. The maximum gamma interferon (IFN gamma) and interleukin-10 (IL-10) mRNA levels were observed on days 11 and 18, respectively, while only negligible amounts of IL-4 and IL-2 were detected throughout the infection. In addition to showing the temporal relationship between treponemal burden and T-cell responses during lesion progression, our results also demonstrate that the composition of the T-cell population changes during lesion resolution. The presence of the mRNA for IFN-gamma, but not IL-4, is consistent with cytokine expression in human syphilis and provides further support for the hypothesis that there is a Th1 predominance during the early immune response to T. pallidum. PMID- 17403877 TI - Disruption of interleukin-27 signaling results in impaired gamma interferon production but does not significantly affect immunopathology in murine schistosome infection. AB - In schistosomiasis mansoni, parasite eggs cause hepatointestinal granulomatous inflammation and fibrosis mediated by CD4 T cells specific for egg antigens. The severity of disease varies extensively in humans and among mouse strains. Marked disease exacerbation induced in typically low-pathology C57BL/6 mice by immunization with schistosome egg antigens (SEA) in complete Freund's adjuvant (SEA/CFA) correlates with elevated production of the proinflammatory cytokines gamma interferon (IFN-gamma) and interleukin-17 (IL-17), which are regulated by IL-12 and IL-23, respectively. Here we examined the effect on the schistosome infection of a third member of the IL-12 family of heterodimeric cytokines, IL 27, using SEA/CFA-immunized and unimmunized mice deficient in the IL-27 receptor chain WSX-1 (WSX-1(-/-)). SEA-stimulated bulk mesenteric lymph node cells or CD4 T cells from 7-week-infected WSX-1(-/-) mice produced significantly less IFN gamma than did those from C57BL/6 mice, even though there was no difference between these mice in exacerbated hepatic egg-induced granulomatous inflammation or in the levels of IL-17 induced by immunization with SEA/CFA. A fraction of the cells in the granulomas stained positive for IL-27, but there were no significant differences between WSX-1(-/-) and BL/6 mice, nor were there differences in the number of CD4 T cells and eosinophils. A 24-week chronic infection resulted in markedly reduced levels of proinflammatory cytokines, including IFN-gamma, in WSX 1(-/-) mice, but again the magnitude of immunopathology was not significantly different between the two groups. These findings indicate that despite the impaired IFN-gamma production, IL-27 signaling has no significant effect on either the magnitude of egg-induced immunopathology or on its closest in vitro correlate, IL-17. PMID- 17403878 TI - Regulation of polysaccharide utilization contributes to the persistence of group a streptococcus in the oropharynx. AB - Group A Streptococcus (GAS) genes that encode proteins putatively involved in polysaccharide utilization show growth phase-dependent expression in human saliva. We sought to determine whether the putative polysaccharide transcriptional regulator MalR influences the expression of such genes and whether MalR helps GAS infect the oropharynx. Analysis of 32 strains of 17 distinct M protein serotypes revealed that MalR is highly conserved across GAS strains. malR transcripts were detectable in patients with GAS pharyngitis, and the levels increased significantly during growth in human saliva compared to the levels during growth in glucose-containing or nutrient-rich media. To determine if MalR influenced the expression of polysaccharide utilization genes, we compared the transcript levels of eight genes encoding putative polysaccharide utilization proteins in the parental serotype M1 strain MGAS5005 and its DeltamalR isogenic mutant derivative. The transcript levels of all eight genes were significantly increased in the DeltamalR strain compared to the parental strain, especially during growth in human saliva. Following experimental infection, the DeltamalR strain persistently colonized the oropharynx in significantly fewer mice than the parental strain colonized, and the numbers of DeltamalR strain CFU recovered were significantly lower than the numbers of the parental strain CFU recovered. These data led us to conclude that MalR influences the expression of genes putatively involved in polysaccharide utilization and that MalR contributes to the persistence of GAS in the oropharynx. PMID- 17403879 TI - Modulation of adherence, invasion, and tumor necrosis factor alpha secretion during the early stages of infection by Streptococcus pneumoniae ClpL. AB - Heat shock proteins (HSPs) play a pivotal role as chaperones in the folding of native and denatured proteins and can help pathogens penetrate host defenses. However, the underlying mechanism(s) of modulation of virulence by HSPs has not been fully determined. In this study, the role of the chaperone ClpL in the pathogenicity of Streptococcus pneumoniae was assessed. A clpL mutant adhered to and invaded nasopharyngeal or lung cells much more efficiently than the wild type adhered to and invaded these cells in vitro, as well as in vivo, although it produced the same amount of capsular polysaccharide. However, the level of secretion of tumor necrosis factor alpha (TNF-alpha) from macrophages infected with the clpL mutant was significantly lower than the level of secretion elicited by the wild type during the early stages of infection. Interestingly, treatment of the human lung epithelial carcinoma A549 and murine macrophage RAW 264.7 cell lines with cytochalasin D, an inhibitor of actin polymerization, increased adherence of the mutant to the host cells. In contrast, cytochalasin D treatment of RAW 264.7 cells decreased TNF-alpha secretion after infection with either the wild type or the mutant. However, pretreatment of cell lines with the actin polymerization activator jasplakinolide reversed these phenotypes. These findings indicate, for the first time, that the ClpL chaperone represses adherence of S. pneumoniae to host cells and induces secretion of TNF-alpha via a mechanism dependent upon actin polymerization during the initial infection stage. PMID- 17403880 TI - Differential innate immune cell activation and proinflammatory response in Anaplasma phagocytophilum infection. AB - Human granulocytic anaplasmosis (HGA) is caused by the obligate intracellular bacterium Anaplasma phagocytophilum. The critical role of gamma interferon (IFN gamma) for induction of severe inflammatory histopathology, even in the absence of a significant bacterial load, was previously demonstrated in a murine model of HGA. We hypothesized that NK, NKT, and possibly CD8(+) cytotoxic T cells participate in the development of histopathologic lesions with A. phagocytophilum infection. Mice were mock infected or infected with low- or high-passage A. phagocytophilum and assayed for hepatic histopathology and splenocyte immunophenotype during the first 21 days after infection. Compared to high passage A. phagocytophilum-infected mice, low-passage A. phagocytophilum-infected mice had more severe hepatic lesions and increased apoptosis. The hepatic histopathology severity in low-passage A. phagocytophilum-infected mice peaked on day 2 at the time of peak plasma IFN-gamma levels and gradually decreased through day 21. Low-passage A. phagocytophilum-infected mice also showed significantly increased levels of lymphocyte NK1.1/FasL expression on days 4 to 7 corresponding to early, severe hepatic inflammation, whereas the levels of NKT cells were substantially lower on day 4, suggesting that there was NKT cell involvement. This result supports the concept that NK1.1(+) cells, including NK and NKT cells, are major components in the early pathogenesis of A. phagocytophilum infection. PMID- 17403881 TI - Salivary duct strictures: nature and incidence in benign salivary obstruction. AB - OBJECTIVES: The aim of this study was to establish the incidence and character of salivary duct strictures by carrying out a 10 year retrospective review. Salivary gland obstruction is most commonly caused either by salivary calculi or duct strictures. These strictures or stenoses develop secondarily to inflammation in the duct wall and may be single or multiple. METHODS: All reports of sialographic examinations performed on patients referred to the Dental Radiology Department in a London Dental Hospital between 1995 and 2004 were reviewed and those patients with symptoms of salivary obstruction identified. In total, 1362 sialograms using the conventional hand injection technique with water-soluble contrast media were performed on 1349 patients with obstructive symptoms during the 10-year period. RESULTS: Of the 1362 sialograms performed, the reports revealed that 877 (64.4%) showed evidence of benign intraductal obstruction. The remaining 485 (35.6%) were normal. 642 of the cases (73.2%) revealing obstruction were reported to be due to salivary calculi, 198 due to duct strictures (22.6%) and the remaining 37 (4.2%) were considered to be due to mucous plugs. Detailed analysis of the patients with strictures showed they were more common in women with a mean age of 52 years. Single strictures were evident in 66.7% of cases while 33.3% showed multiple stenoses. Strictures were more common in the parotid duct (75.3%). 7% of patients presented with bilateral stenoses. CONCLUSION: This is the largest review of duct strictures to be reported. It has shown that ductal stricture formation accounts for almost 25% of cases of benign salivary obstruction and appears to have been an under-recognized condition. Strictures more commonly affect parotid ducts and are typically found in the fourth, fifth or sixth decades, particularly in women. PMID- 17403882 TI - Diagnostic accuracy of in vitro panoramic radiographs depending on the exposure. AB - OBJECTIVES: The aim was to evaluate the effect of dose reduction on diagnostic accuracy in panoramic radiographs with increased tube potential and reduced milliampere settings. METHODS: Panoramic radiographs of 12 dried human skulls prepared with lesions in the bone, teeth and peri-implant bone in ascending size were taken. Medium and regular film--screen combinations and a storage phosphor system were used for imaging. All systems were exposed at a low and a high tube potential level. To compare the dose at different tube potential settings, dose length product was measured at the secondary collimator. Five observers assessed the presence (response: 1) or absence (response: 0) of lesions. Sensitivity, specificity and diagnostic accuracy were evaluated and 36 000 ratings were made in all. All settings were repeated once. Intrarater agreement was expressed by Cohen's kappa coefficient. RESULTS: There was no significant difference in diagnostic accuracy between a medium and a regular film--screen combination at a low tube potential level (70 kV; 0.935 and 0.930) and the medium film--screen system at a high tube potential level (85 kV; 0.926). Compared with this group, the regular film-screen combinations at high tube potential level (85 kV, 0.906) and all digital radiographs were significantly different (0.886 and 0.866), irrespective of the tube potential level. The digital panoramic radiograph was only comparable with the best film--screen combinations with an exposure for a medium film-screen system and at a low tube potential level. Sensitivity was 89.9% and specificity 93.7%. The kappa coefficient for intrarater agreement was high (0.81). CONCLUSIONS: The medium intensifying screen can be used at high tube potential settings instead of low tube potential settings, or the regular intensifying screen can be used at low tube potential settings with the same diagnostic value. A dose reduction of about 40% is possible. The storage phosphor plates should be exposed at least like a regular film-screen system and at a low tube potential level. PMID- 17403883 TI - Influence of tube potential setting and dose on the visibility of lesions in intraoral radiography. AB - OBJECTIVES: The objectives were to determine the diagnostic accuracy of different image receptors when first, the tube potential setting was increased from 60 kV to 90 kV and second, when the milliampere setting was reduced at unchanged tube potential for storage phosphor plates. METHODS: Intraoral radiographs (films of speed class F, storage phosphor plates) were taken of 12 dried human skulls prepared with lesions in both teeth and peri-implant bone, in ascending size. Five observers assessed the presence (response=1) or absence (response=0) of lesions. The digital intraoral radiograph was exposed at 60 kV and 90 kV with half and a quarter of the film exposure. Some of the radiographs were assessed a second time. In total, 6000 assessments were made. Intrarater agreement was expressed by Cohen's kappa coefficient. RESULTS: The digital combinations showed the lowest diagnostic accuracy in all combinations, but the results demonstrated an equivalence of all combinations of films and storage phosphor plates. The differences in diagnostic accuracy were low (94-96.5%). The kappa coefficient for intrarater agreement was high (0.85). CONCLUSIONS: Regarding peri-implant and decayed lesions, intraoral films and storage phosphor plates demonstrated equal quality in this in vitro study regardless of exposure at 60 kV or 90 kV. PMID- 17403884 TI - The 3DX multi image micro-CT device in clinical dental practice. AB - OBJECTIVES: To evaluate the usefulness of 3DX multi-image micro-CT device in clinical dental practice. METHODS: Images of 198 examinations performed with a 3DX device during a 6 month period in a private dental practice were evaluated retrospectively for the benefit of additional information compared with panoramic or intraoral radiographs. RESULTS: The main indication for the use of 3DX micro CT was planning of dental implant placement in 49% of the examinations. Diagnosis or exclusion of dental infection or peri-implantitis represented 28% of the examinations and tooth, root or foreign body localization represented 13%. Temporomandibular joint (TMJ) imaging and cyst or tumour diagnosis represented 7.5% and 2.5% of the examinations, respectively. In implant planning and tooth, root or foreign body localization examinations, the required information was obtained in every case except three implant planning examinations, in which the exact measurements were hampered by artefacts caused by root fillings and retrograde fillings or metal posts. When compared with conventional radiography, additional radiographic information was obtained in 51% of the micro-CT examinations performed to confirm or exclude dental infection or peri implantitis. Micro-CT also showed the bone structure of TMJs precisely. CONCLUSIONS: The 3DX micro-CT device visualizes bony anatomical structures precisely, which makes it a reliable tool for, for example, planning of implant treatment. The device is highly suitable for diagnosing dental infections. Dental restorations may cause disturbing artefacts and in 4.5% of the examinations the small imaging area resulted in re-examination. PMID- 17403885 TI - Visual-spatial ability and interpretation of three-dimensional information in radiographs. AB - OBJECTIVES: To investigate whether skill in the interpretation of three dimensional (3D) information in radiographs utilizing the parallax phenomenon is associated with visual-spatial ability and whether development of this skill is related to visual-spatial ability. METHODS: Eighty-six individuals with a median age of 25 years participated in the study. It was organized into three parts: (1) assessment before training, (2) training in object depth localization utilizing parallax and (3) assessment after training. Before training, visual-spatial ability was assessed with a mental rotation test, MRT-A; skill in interpreting 3D information was assessed with two specifically designed proficiency tests: a radiography test, which assessed the ability to interpret 3D information in radiographs utilizing motion parallax and a principle test which assessed understanding of the principles of motion parallax. After training, skill in interpreting 3D information was reassessed. Improvement was defined as the difference between test scores after training and before training. Multiple linear regression was used to analyse the effect of student and training characteristics on proficiency test results and improvement. RESULTS: Radiography test results after training and improvement of radiography test results were significantly associated with MRT-A scores (P<0.001 and P=0.020, respectively). Principle test results were high before training and did not improve after training. The test results were associated with MRT-A both before (P=0.009) and after training (P=0.003). CONCLUSIONS: Understanding of the parallax phenomenon is associated with visual-spatial ability. Development of the skill to interpret 3D information in radiographs utilizing parallax is facilitated for individuals with high visual-spatial ability. PMID- 17403886 TI - Carotid artery calcification seen on panoramic dental radiographs in the Asian population in Japan. AB - OBJECTIVES: To determine the frequency of carotid artery calcification (CAC) seen on panoramic dental radiographs, the relationship between the condition and a history of cerebral infarction (CI), and the history of the risk factors of CI in patients in Japan. METHODS: A total of 2374 individuals whose panoramic radiographs were obtained at their first consultation were reviewed. RESULT: CAC was found in 95 subjects (4.0%) and was found at a higher frequency in patients with a history of CI than in those without a history of CI. The modifiable risk factors (a history of hypertension, diabetes and hypercholesterolaemia) and CAC showed significant relationship in the chi(2) test for pairwise comparison. When age and gender were added and all of these risk factors were considered simultaneously in the multiple logistic regression analysis, age and history of hypercholesterolaemia remained statistically significant. CONCLUSIONS: CAC may be seen on panoramic dental radiographs and is related to the patients' general history of CI, and the risk factors of CI in the Asian population in Japan. PMID- 17403887 TI - Evaluation of optical density of the midpalatal suture 3 months after surgically assisted rapid maxillary expansion. AB - OBJECTIVES: This study evaluated new bone formation at the midpalatal suture after surgically assisted rapid maxillary expansion (SARME) by optical density analysis. METHODS: The study population consisted of 18 patients, 10 males and 8 females. All patients presented maxillary atresia with posterior crossbite and were submitted to SARME. Maxillary occlusal radiographs were taken at three stages (before SARME, immediately after SARME and after 3 months). Three patients did not attend the session at the 3-month period for achievement of the occlusal radiography and thus were excluded, leading to a final sample of 15 patients. Two regions were selected and analysed at each stage. Region A measured 8 x 1 mm(2) and was located 1.2 cm from the tangent to the maxillary central incisors at the region of the midpalatal suture. Region B measured 5 x 9 mm(2) and was located 4.3 cm from the tangent to the maxillary central incisors at the region of the midpalatal suture. An aluminium scale (step wedge) with eight steps varying from 1 mm to 8 mm was adapted at the end of the films. Radiographs were taken on a Spectro 70X machine (Dabi Atlante, Ribeirao Preto, Brazil) set at 70 kVp and 10 mA with an exposure time of 1.0 s. Radiographs were scanned using a Power Look 1000 scanner (Umax, Taiwan, China) and a computer Dimension E510 (Dell Computer, Taiwan, China). Optical density analysis was performed after digitization of radiographs using the software Image Tool (UTHSCSA, San Antonio, TX). RESULTS: Statistical analysis of region A revealed statistically significant differences between Stages I and II (P=0.0001), Stages II and III (P=0.0001) and Stages I and III (P=0.0003). In region B, statistically significant differences were observed between Stages I and II, I and III and II and III (P=0.0018, P=0.0003 and P=0.0003, respectively). CONCLUSIONS: Optical density analysis improves post treatment control of SARME procedures by surgeons and orthodontists. After 3 months, new bone formation at the midpalatal suture is not complete. PMID- 17403888 TI - Chronic invasive sinus aspergillosis in an immunocompetent patient: a case report. AB - We report the case of an immunocompetent, 39 year old male who presented with diplopia and diffuse headache. The CT scan demonstrated the presence of a sizeable mass that took over the nasal cavity, the right maxillary sinus, the sphenoid and ethmoid sinus, extending to the sellar and suprasellar region, and eroding the adjacent bones. The patient also underwent MRI and endoscopic sinus surgery. Biopsy of the suspected tissue for pathology and culture demonstrated Aspergillus fumigatus. PMID- 17403889 TI - Pneumosinus dilatans of the ethmoid sinus presenting with exophthalmus: a case report and review of the literature. AB - Pneumosinus dilatans (PSD) is a rare condition of unknown aetiology in which there is enlargement of the paranasal sinuses by air, where the affected sinus expands beyond the normal boundaries of bone without thinning of its bony walls. The case of a 28 year old male with PSD, who had gradually increasing exophthalmus and diplopia, is presented and characteristics of this condition are reviewed with literature data. PMID- 17403890 TI - Intracranial aneurysm as an incidental finding. AB - This report presents an incidental finding of a potentially life-threatening large fusiform aneurysm of the internal carotid artery at the level of the posterior communicating artery in a patient who reported with complaints relating to the presence of a radiolucent lesion in the mandible that was diagnosed as odontogenic keratocyst. This case highlights the need to study a case in detail in preparation for prescribing the appropriate advanced imaging modality examination, and also to carefully evaluate the entire imaged volume for incidental pathology and not just the region of interest. This report assumes significance in the light of widespread use of cone beam CT by dental clinicians for routine diagnostic tasks without a formal interpretation being carried out on all such studies. PMID- 17403891 TI - CT and MRI findings of intraosseous schwannoma of the mandible: a case report. AB - We report the radiographical findings of a rare case of intraosseous schwannoma of the mandible. The tumour that presented as a unilocular, well-defined, radiolucent lesion on plain radiography was located in the molar region. On CT, the tumour was a well-demarcated mass with no periosteal reaction and no destruction of the bone cortex. Destructive changes in the cortical wall of the mandibular canal by the tumour were observed on CT, but no evidence of dilatation and shift in the inferior mandibular canal was seen. MR imaging revealed that the mandibular canal was encased by the tumour as a solid mass without cystic parts. The signal intensity of the tumour was non-specific on MRI. Characteristics of intraosseous schwannoma in the mandible are the encasement of the canal by a well demarcated tumour without periosteal reaction and the destruction of mandibular bone cortex. The destructive change of the inferior mandibular canal can be observed on CT and MRI. However, a biopsy is necessary to make the final diagnosis because of the non-specificity of the findings. PMID- 17403892 TI - Extracranial meningioma of the parapharyngeal space: report of a case and review of the literature. AB - Parapharyngeal tumours are frequently encountered in clinical practice and are often difficult to diagnose upon clinical examination due to the anatomic complexity of the region. We report a rare case of extracranial meningioma of the parapharyngeal space in a patient with no history of von Recklinghausen's disease, family history of meningioma or central neurofibromatosis (NF-2). The purpose of this report is to reiterate and emphasize the role of CT and MRI in the diagnosis of parapharyngeal masses, and to describe the characteristic radiological features of extracranial meningiomas. PMID- 17403893 TI - Radiographic assessment of Gardner's syndrome. AB - The detection of osteomas in the maxillofacial region may be the initial clinical finding in Gardner's syndrome (GS). The most common location of osteomas is in the skull, but the lesion can also occur in the jaws. We present a case of a 47 year old male patient with GS who was referred for radiological evaluation. Extraoral examination revealed an epidermoid cyst and the patient had a history of intestinal polyps. A panoramic radiograph demonstrated numerous osteomas and diffuse sclerosis of the mandible, and compound odontomas with impacted teeth. CT scan allowed the localization and extension of the osteomas, and showed other sites in the maxillofacial region as well. CT images also revealed a different behaviour of osteoma, invading the mandibular canal. PMID- 17403894 TI - The ternary complex factor net is downregulated by hypoxia and regulates hypoxia responsive genes. AB - Hypoxia and the Net ternary complex factor (TCF) regulate similar processes (angiogenesis, wound healing, and cellular migration) and genes (PAI-1, c-fos, erg-1, NOS-2, HO-1, and vascular endothelial growth factor genes), suggesting that they are involved in related pathways. We show here that hypoxia regulates Net differently from the other TCFs and that Net plays a role in the hypoxic response in vivo in mice and in cells. Hypoxia induces Net depletion from target promoters, nuclear export, ubiquitylation, and proteasomal degradation. Key mediators of the hypoxic response, the prolyl-4-hydroxylases containing domain proteins (PHDs), regulate Net. PHD downregulation in normoxia leads to Net degradation, and PHD overexpression delays Net downregulation by hypoxia. Net inhibition by RNA interference or mutation leads to altered regulation by hypoxia of the Net targets PAI-1, c-fos, and egr-1. We propose that hypoxia stimulates transcription of target promoters through removal of the repressor function of Net. Interestingly, the hematocrit response to a chemical inducer of hypoxia-like responses (cobalt chloride) is strongly altered in Net mutant mice. Our results show that the Net TCF is part of the biological response to hypoxia, adding a new component to an important pathological and physiological process. PMID- 17403895 TI - Chibby promotes adipocyte differentiation through inhibition of beta-catenin signaling. AB - The canonical Wnt/beta-catenin signaling pathway plays diverse roles in embryonic development and disease. Activation of this pathway, likely by Wnt-10b, has been shown to inhibit adipogenesis in cultured 3T3-L1 preadipocytes and in mice. Here, we report that the beta-catenin antagonist Chibby (Cby) is required for adipocyte differentiation. Cby is expressed in adipose tissue in mice, and Cby protein levels increase during adipogenic differentiation of 3T3-L1 cells. Ectopic expression of Cby induces spontaneous differentiation of these cells into mature adipocytes to an extent similar to that of dominant-negative Tcf-4. In contrast, depletion of Cby by RNA interference potently blocks adipogenesis of 3T3-L1 and mouse embryonic stem cells. In support of this, embryonic fibroblasts obtained from Cby-deficient embryos display attenuated differentiation to the adipogenic lineage. Mechanistically, Cby promotes adipocyte differentiation, in part by inhibiting beta-catenin, since gain or loss of function of Cby influences beta catenin signaling in 3T3-L1 cells. Our results therefore establish Cby as a novel proadipogenic factor required for adipocyte differentiation. PMID- 17403896 TI - Eos, MITF, and PU.1 recruit corepressors to osteoclast-specific genes in committed myeloid progenitors. AB - Transcription factors MITF and PU.1 collaborate to increase expression of target genes like cathepsin K (Ctsk) and acid phosphatase 5 (Acp5) during osteoclast differentiation. We show that these factors can also repress transcription of target genes in committed myeloid precursors capable of forming either macrophages or osteoclasts. The direct interaction of MITF and PU.1 with the zinc finger protein Eos, an Ikaros family member, was necessary for repression of Ctsk and Acp5. Eos formed a complex with MITF and PU.1 at target gene promoters and suppressed transcription through recruitment of corepressors CtBP (C-terminal binding protein) and Sin3A, but during osteoclast differentiation, Eos association with Ctsk and Acp5 promoters was significantly decreased. Subsequently, MITF and PU.1 recruited coactivators to these target genes, resulting in robust expression of target genes. Overexpression of Eos in bone marrow-derived precursors disrupted osteoclast differentiation and selectively repressed transcription of MITF/PU.1 targets, while small interfering RNA knockdown of Eos resulted in increased basal expression of Ctsk and Acp5. This work provides a mechanism to account for the modulation of MITF and PU.1 activity in committed myeloid progenitors prior to the initiation of osteoclast differentiation in response to the appropriate extracellular signals. PMID- 17403897 TI - The ancient mariner sails again: transposition of the human Hsmar1 element by a reconstructed transposase and activities of the SETMAR protein on transposon ends. AB - Hsmar1, one of the two subfamilies of mariner transposons in humans, is an ancient element that entered the primate genome lineage approximately 50 million years ago. Although Hsmar1 elements are inactive due to mutational damage, one particular copy of the transposase gene has apparently been under selection. This transposase coding region is part of the SETMAR gene, in which a histone methylatransferase SET domain is fused to an Hsmar1 transposase domain. A phylogenetic approach was taken to reconstruct the ancestral Hsmar1 transposase gene, which we named Hsmar1-Ra. The Hsmar1-Ra transposase efficiently mobilizes Hsmar1 transposons by a cut-and-paste mechanism in human cells and zebra fish embryos. Hsmar1-Ra can also mobilize short inverted-repeat transposable elements (MITEs) related to Hsmar1 (MiHsmar1), thereby establishing a functional relationship between an Hsmar1 transposase source and these MITEs. MiHsmar1 excision is 2 orders of magnitude more efficient than that of long elements, thus providing an explanation for their high copy numbers. We show that the SETMAR protein binds and introduces single-strand nicks into Hsmar1 inverted-repeat sequences in vitro. Pathway choices for DNA break repair were found to be characteristically different in response to transposon cleavage mediated by Hsmar1-Ra and SETMAR in vivo. Whereas nonhomologous end joining plays a dominant role in repairing excision sites generated by the Hsmar1-Ra transposase, DNA repair following cleavage by SETMAR predominantly follows a homology-dependent pathway. The novel transposon system can be a useful tool for genome manipulations in vertebrates and for investigations into the transpositional dynamics and the contributions of these elements to primate genome evolution. PMID- 17403898 TI - Selective requirement for SAGA in Hog1-mediated gene expression depending on the severity of the external osmostress conditions. AB - Regulation of gene expression by the Hog1 stress-activated protein kinase is essential for proper cell adaptation to osmostress. Hog1 coordinates an extensive transcriptional program through the modulation of transcription. To identify systematically novel components of the transcriptional machinery required for osmostress-mediated gene expression, we performed an exhaustive genome-wide genetic screening, searching for mutations that render cells osmosensitive at high osmolarity and that are associated with reduced expression of osmoresponsive genes. The SAGA and Mediator complexes were identified as putative novel regulators of osmostress-mediated transcription. Interestingly, whereas Mediator is essential for osmostress gene expression, the requirement for SAGA is different depending on the strength of the extracellular osmotic conditions. At mild osmolarity, SAGA mutants show only very slight defects on RNA polymerase II (Pol II) recruitment and gene expression, whereas at severe osmotic conditions, SAGA mutants show completely impaired RNA Pol II recruitment and transcription of osmoresponsive genes. Thus, our results define an essential role for Mediator in osmostress gene expression and a selective role for SAGA under severe osmostress. Our results indicate that the requirement for a transcriptional complex to regulate a promoter might be determined by the strength of the stimuli perceived by the cell through the regulation of interactions between transcriptional complexes. PMID- 17403899 TI - CIF-1, a shared subunit of the COP9/signalosome and eukaryotic initiation factor 3 complexes, regulates MEL-26 levels in the Caenorhabditis elegans embryo. AB - The COP9/signalosome (CSN) is an evolutionarily conserved macromolecular complex that regulates the cullin-RING ligase (CRL) class of E3 ubiquitin ligases, primarily by removing the ubiquitin-like protein Nedd8 from the cullin subunit. In the Caenorhabditis elegans embryo, the CSN controls the degradation of the microtubule-severing protein MEI-1 through CUL-3 deneddylation. However, the molecular mechanisms of CSN function and its subunit composition remain to be elucidated. Here, using a proteomic approach, we have characterized the CSN and CUL-3 complexes from C. elegans embryos. We show that the CSN physically interacts with the CUL-3-based CRL and regulates its activity by counteracting the autocatalytic instability of the substrate-specific adaptor MEL-26. Importantly, we identified the uncharacterized protein K08F11.3/CIF-1 (for CSN eukaryotic initiation factor 3 [eIF3]) as a stoichiometric and functionally important subunit of the CSN complex. CIF-1 appears to be the only ortholog of Csn7 encoded by the C. elegans genome, but it also exhibits extensive sequence similarity to eIF3m family members, which are required for the initiation of protein translation. Indeed, CIF-1 binds eIF-3.F and inactivation of cif-1 impairs translation in vivo. Taken together, our results indicate that CIF-1 is a shared subunit of the CSN and eIF3 complexes and may therefore link protein translation and degradation. PMID- 17403900 TI - Cooperative interaction between hepatocyte nuclear factor 4 alpha and GATA transcription factors regulates ATP-binding cassette sterol transporters ABCG5 and ABCG8. AB - Cholesterol homeostasis is maintained by coordinate regulation of cholesterol synthesis and its conversion to bile acids in the liver. The excretion of cholesterol from liver and intestine is regulated by ATP-binding cassette half transporters ABCG5 and ABCG8. The genes for these two proteins are closely linked and divergently transcribed from a common intergenic promoter region. Here, we identified a binding site for hepatocyte nuclear factor 4alpha (HNF4alpha) in the ABCG5/ABCG8 intergenic promoter, through which HNF4alpha strongly activated the expression of a reporter gene in both directions. The HNF4alpha-responsive element is flanked by two conserved GATA boxes that were also required for stimulation by HNF4alpha. GATA4 and GATA6 bind to the GATA boxes, coexpression of GATA4 and HNF4alpha leads to a striking synergistic activation of both the ABCG5 and the ABCG8 promoters, and binding sites for HNF4alpha and GATA were essential for maximal synergism. We also show that HNF4alpha, GATA4, and GATA6 colocalize in the nuclei of HepG2 cells and that a physical interaction between HNF4alpha and GATA4 is critical for the synergistic response. This is the first demonstration that HNF4alpha acts synergistically with GATA factors to activate gene expression in a bidirectional fashion. PMID- 17403901 TI - Ptf1a binds to and activates area III, a highly conserved region of the Pdx1 promoter that mediates early pancreas-wide Pdx1 expression. AB - The critical pancreatic transcription factor Pdx1 is expressed throughout the pancreas early but enriched in insulin-producing beta cells postnatally. Previous studies showed that the 5' conserved promoter regions areas I and II (Pdx1(PB)) direct endocrine cell expression, while an adjacent region (Pdx1(XB)) containing conserved area III directs transient beta-cell expression. In this study, we used Cre-mediated lineage tracing to track cells that activated these regions. Pdx1(PB)Cre mediated only endocrine cell recombination, while Pdx1(XB)Cre directed broad and early recombination in the developing pancreas. Also, a reporter transgene containing areas I, II, and III was expressed throughout the embryonic day 10.5 (E10.5) pancreas and gradually became beta cell enriched, similar to endogenous Pdx1. These data suggested that sequences within area III mediate early pancreas-wide Pdx1 expression. Area III contains a binding site for PTF1, a transcription factor complex essential for pancreas development. This site contributed to area III-dependent reporter gene expression in the acinar AR42J cell line, while PTF1 specifically trans-activated area III-containing reporter expression in a nonpancreatic cell line. Importantly, Ptf1a occupied sequences spanning the endogenous PTF1 site in area III of E11.5 pancreatic buds. These data strongly suggest that PTF1 is an important early activator of Pdx1 in acinar and endocrine progenitor cells during pancreas development. PMID- 17403902 TI - Upregulation of Twist-1 by NF-kappaB blocks cytotoxicity induced by chemotherapeutic drugs. AB - NF-kappaB/Rel transcription factors are central to controlling programmed cell death (PCD). Activation of NF-kappaB blocks PCD induced by numerous triggers, including ligand engagement of tumor necrosis factor receptor (TNF-R) family receptors. The protective activity of NF-kappaB is also crucial for oncogenesis and cancer chemoresistance. Downstream of TNF-Rs, this activity of NF-kappaB has been linked to the suppression of reactive oxygen species and the c-Jun-N terminal-kinase (JNK) cascade. The mechanism by which NF-kappaB inhibits PCD triggered by chemotherapeutic drugs, however, remains poorly understood. To understand this mechanism, we sought to identify unrecognized protective genes that are regulated by NF-kappaB. Using an unbiased screen, we identified the basic-helix-loop-helix factor Twist-1 as a new mediator of the protective function of NF-kappaB. Twist-1 is an evolutionarily conserved target of NF kappaB, blocks PCD induced by chemotherapeutic drugs and TNF-alpha in NF-kappaB deficient cells, and is essential to counter this PCD in cancer cells. The protective activity of Twist-1 seemingly halts PCD independently of interference with cytotoxic JNK, p53, and p19(ARF) signaling, suggesting that it mediates a novel protective mechanism activated by NF-kappaB. Indeed, our data indicate that this activity involves a control of inhibitory Bcl-2 phosphorylation. The data also suggest that Twist-1 and -2 play an important role in NF-kappaB-dependent chemoresistance. PMID- 17403903 TI - Depletion of the yeast nuclear exosome subunit Rrp6 results in accumulation of polyadenylated RNAs in a discrete domain within the nucleolus. AB - Recent data reveal that a substantial fraction of transcripts generated by RNA polymerases I, II, and III are rapidly degraded in the nucleus by the combined action of the exosome and a noncanonical poly(A) polymerase activity. This work identifies a domain within the yeast nucleolus that is enriched in polyadenylated RNAs in the absence of the nuclear exosome RNase Rrp6 or the exosome cofactor Mtr4. In normal yeast cells, poly(A)(+) RNA was undetectable in the nucleolus but the depletion of either Rrp6 or Mtr4 led to the accumulation of polyadenylated RNAs in a discrete subnucleolar region. This nucleolar poly(A) domain is enriched for the U14 snoRNA and the snoRNP protein Nop1 but is distinct from the nucleolar body that functions in snoRNA maturation. In strains lacking both Rrp6 and the poly(A) polymerase Trf4, the accumulation of poly(A)(+) RNA was suppressed, suggesting the involvement of the Trf4-Air1/2-Mtr4 polyadenylation (TRAMP) complex. The accumulation of polyadenylated snoRNAs in a discrete nucleolar domain may promote their recognition as substrates for the exosome. PMID- 17403904 TI - RIAM links the ADAP/SKAP-55 signaling module to Rap1, facilitating T-cell receptor-mediated integrin activation. AB - One outcome of T-cell receptor (TCR) signaling is increased affinity and avidity of integrins for their ligands. This occurs through a process known as inside-out signaling, which has been shown to require several molecular components including the adapter proteins ADAP (adhesion and degranulation-promoting adapter protein) and SKAP-55 (55-kDa src kinase-associated phosphoprotein) and the small GTPase Rap1. Herein, we provide evidence linking ADAP and SKAP-55 to RIAM, a recently described adapter protein that binds selectively to active Rap1. We identified RIAM as a key component linking the ADAP/SKAP-55 module to the small GTPase Rap1, facilitating TCR-mediated integrin activation. We show that RIAM constitutively interacts with SKAP-55 in both a heterologous transfection system and primary T cells and map the region essential for this interaction. Additionally, we find that the SKAP-55/RIAM complex is essential both for TCR-mediated adhesion and for efficient conjugate formation between T cells and antigen-presenting cells. Mechanistic studies revealed that the ADAP/SKAP-55 module relocalized RIAM and Rap1 to the plasma membrane following TCR activation to facilitate integrin activation. These results describe for the first time a link between ADAP/SKAP-55 and the Rap1/RIAM complex and provide a potential new mechanism for TCR-mediated integrin activation. PMID- 17403905 TI - Conformational modification of serpins transforms leukocyte elastase inhibitor into an endonuclease involved in apoptosis. AB - The best-characterized biochemical feature of apoptosis is degradation of genomic DNA into oligonucleosomes. The endonuclease responsible for DNA degradation in caspase-dependent apoptosis is caspase-activated DNase. In caspase-independent apoptosis, different endonucleases may be activated according to the cell line and the original insult. Among the known effectors of caspase-independent cell death, L-DNase II (LEI [leukocyte elastase inhibitor]-derived DNase II) has been previously characterized by our laboratory. We have thus shown that this endonuclease derives from the serpin superfamily member LEI by posttranslational modification (A. Torriglia, P. Perani, J. Y. Brossas, E. Chaudun, J. Treton, Y. Courtois, and M. F. Counis, Mol. Cell. Biol. 18:3612-3619, 1998). In this work, we assessed the molecular mechanism involved in the change in the enzymatic activity of this molecule from an antiprotease to an endonuclease. We report that the cleavage of LEI by elastase at its reactive center loop abolishes its antiprotease activity and leads to a conformational modification that exposes an endonuclease active site and a nuclear localization signal. This represents a novel molecular mechanism for a complete functional conversion induced by changing the conformation of a serpin. We also show that this molecular transformation affects cellular fate and that both endonuclease activity and nuclear translocation of L-DNase II are needed to induce cell death. PMID- 17403906 TI - P-body formation is a consequence, not the cause, of RNA-mediated gene silencing. AB - P bodies are cytoplasmic domains that contain proteins involved in diverse posttranscriptional processes, such as mRNA degradation, nonsense-mediated mRNA decay (NMD), translational repression, and RNA-mediated gene silencing. The localization of these proteins and their targets in P bodies raises the question of whether their spatial concentration in discrete cytoplasmic domains is required for posttranscriptional gene regulation. We show that processes such as mRNA decay, NMD, and RNA-mediated gene silencing are functional in cells lacking detectable microscopic P bodies. Although P bodies are not required for silencing, blocking small interfering RNA or microRNA silencing pathways at any step prevents P-body formation, indicating that P bodies arise as a consequence of silencing. Consistently, we show that releasing mRNAs from polysomes is insufficient to trigger P-body assembly: polysome-free mRNAs must enter silencing and/or decapping pathways to nucleate P bodies. Thus, even though P-body components play crucial roles in mRNA silencing and decay, aggregation into P bodies is not required for function but is instead a consequence of their activity. PMID- 17403907 TI - Combinatorial expression of alpha- and gamma-protocadherins alters their presenilin-dependent processing. AB - Alpha- and gamma-protocadherins (Pcdhs) are type I transmembrane receptors expressed predominantly in the central nervous system and located in part in synapses. They are transcribed from complex genomic loci, giving rise in the mouse to 14 alpha-Pcdh and 22 gamma-Pcdh isoforms consisting of variable domains, each encompassing the extracellular region, the transmembrane region, and part of the intracellular region harboring the alpha- or gamma-Pcdh-specific invariant cytoplasmic domain. Presenilin-dependent intramembrane proteolysis (PS-IP) of gamma-Pcdhs and the formation of alpha/gamma-Pcdh heteromers led us to investigate the effects of homo- and heteromer formation on gamma- and putative alpha-Pcdh membrane processing and signaling. We find that upon surface delivery, alpha-Pcdhs, like gamma-Pcdhs, are subject to matrix metallo-protease cleavage followed by PS-IP in neurons. We further demonstrate that the combinatorial expression of alpha- and gamma-Pcdhs modulates the extent of their PS-IP, indicating the formation of alpha/gamma-Pcdh heteromers with an altered susceptibility to processing. Cell-specific expression of alpha/gamma-Pcdh isoforms could thus determine cell and synapse adhesive properties as well as intracellular and nuclear signaling by their soluble cytoplasmic cleavage products, alpha C-terminal fragment 2 (alpha-CTF-2) and gamma-CTF-2. PMID- 17403908 TI - Methodologic issues regarding the use of three observational study designs to assess influenza vaccine effectiveness. AB - BACKGROUND: Influenza causes substantial morbidity and annual vaccination is the most important prevention strategy. Accurately measuring vaccine effectiveness (VE) is difficult. The clinical syndrome most closely associated with influenza virus infection, influenza-like illness (ILI), is not specific. In addition, laboratory confirmation is infrequently done, and available rapid diagnostic tests are imperfect. The objective of this study was to estimate the joint impact of rapid diagnostic test sensitivity and specificity on VE for three types of study designs: a cohort study, a traditional case-control study, and a case control study that used as controls individuals with ILI who tested negative for influenza virus infection. METHODS: We developed a mathematical model with five input parameters: true VE, attack rates (ARs) of influenza-ILI and non-influenza ILI and the sensitivity and specificity of the diagnostic test. RESULTS: With imperfect specificity, estimates from all three designs tended to underestimate true VE, but were similar except if fairly extreme inputs were used. Only if test specificity was 95% or more or if influenza attack rates doubled that of background illness did the case-control method slightly overestimate VE. The case control method usually produced the highest and most accurate estimates, followed by the test-negative design. The bias toward underestimating true VE introduced by low test specificity increased as the AR of influenza- relative to non influenza-ILI decreases and, to a lesser degree, with lower test sensitivity. CONCLUSIONS: Demonstration of a high influenza VE using tests with imperfect sensitivity and specificity should provide reassurance that the program has been effective in reducing influenza illnesses, assuming adequate control of confounding factors. PMID- 17403909 TI - Brief report: a brief intervention to improve lifeguard surveillance at a public swimming pool. AB - OBJECTIVES: Drowning is the second leading cause of unintentional death for American children in middle childhood, but behavioral research designed to prevent pediatric drowning is limited. This study tested the efficacy of a brief intervention to improve lifeguard attention and surveillance at a public swimming pool. METHOD: Observational data on patron risk-taking and lifeguard attention, distraction, and scanning were collected at a public swimming pool, both before and after a brief intervention. The intervention was designed to increase lifeguards' perception of susceptibility of drowning incidents, educate about potential severity of drowning, and help overcome perceived barriers about scanning the pool. RESULTS: Postintervention, lifeguards displayed better attention and scanning and patrons displayed less risky behavior. Change was maintained for the remainder of the season. CONCLUSION: Theoretically driven brief interventions targeting lifeguard attention and surveillance might prove effective in reducing risk of drowning in public swimming pools. PMID- 17403910 TI - Trajectories of adjustment in mothers of children with newly diagnosed cancer: a natural history investigation. AB - OBJECTIVES: The objectives of this study were (a) to assess negative affectivity and posttraumatic symptomatology in mothers following the diagnosis of cancer in their children; (b) to examine sociodemographic and psychosocial variables associated with change in distress over time; and (c) to identify distinct subgroups of mothers whose patterns and trajectories of adjustment can be distinguished according to available predictor data. METHODS: Two hundred and twelve mothers at seven sites were assessed just following their child's diagnosis, and again 3 months and 6 months later. Primary outcomes included measures of mood disturbance, depressive symptoms, and symptoms of posttraumatic stress. RESULTS: Overall, mothers demonstrated a pattern of mildly elevated negative affectivity and posttraumatic symptomatology initially, with steady improvements evident at 3- and 6-month follow-up. Distinct adjustment trajectories were evident within the sample as a whole, indicating subgroups of mothers with high-declining, moderate-stable, and low-stable distress levels. CONCLUSIONS: These findings highlight considerable resilience among mothers facing the stress of childhood cancer. Intervention efforts aimed at reducing maternal distress might best be targeted towards the subgroup of mothers who may be predicted to exhibit the highest level of distress. PMID- 17403911 TI - Factors associated with retaliatory attitudes among African American adolescents who have been assaulted. AB - OBJECTIVES: (a) To describe attitudes regarding retaliation among adolescents who have been assaulted. (b) To examine assault/event characteristics, personal, parental, and environmental factors associated with the retaliatory attitudes of adolescents who have been assaulted. METHODS: African American youth aged 10-15 years presenting to two large urban hospitals with peer assault injury and a parent/caregiver completed interviews in their home after their emergency department visit. RESULTS: Multivariate analyses revealed that lower SES, older age, and adolescents' perceptions that their parents support fighting were related to endorsing retaliatory attitudes. Girls who were aggressive were more likely to endorse retaliatory attitudes. However, level of aggression did not impact boys' retaliatory attitudes. Affiliating with aggressive peers influenced the retaliatory attitudes of boys, but did not influence girls' retaliatory attitudes. Overall, youths' perceptions of their parents' attitudes toward fighting had the greatest impact on retaliatory attitudes. CONCLUSIONS: Adolescents' perceptions of their parents' attitudes toward fighting may be a factor in subsequent re-injury among youth. Violence prevention and intervention efforts need to involve components that assess parental attitudes and incorporate strategies to engage parents in violence prevention efforts. In addition, interventions for youth who have been assaulted may need to incorporate some gender-specific components in order to address the unique needs of girls and boys. PMID- 17403912 TI - Patient-reported outcomes in end-of-life research in pediatric oncology. AB - OBJECTIVE: The purpose of this review of published literature was to identify the number and focus of empirically based papers that included research methods used to directly solicit patient-reported outcomes (PRO) from pediatric oncology patients at end of life. METHODS: Key terms including "pediatric or child and oncology or cancer and end of life or palliative or hospice or dying" were used with five data bases (PubMed, Ovid, Cochrane, PsycInfo & PsycArticles, and CINAHL) for English language literature published between January, 2001 and June, 2006. All retrieved documents were independently reviewed by a panel of six (nurses, physicians, and one psychologist) with backgrounds in pediatric oncology. RESULTS: Thirty-five publications were identified but nine (25.7%) were eliminated from the analysis as they did not meet inclusion criteria. Of the remaining 26, four (15.4%) included patient-reported outcomes, six (23.1%) included parent only-reported outcomes, and five (19.2%) included staff only reported outcomes. Nine (34.6%) were retrospective medical record reviews. Two (7.7%) included parent and record review data or parent and physician reports. CONCLUSIONS: Empirically-based end-of-life publications in pediatric oncology are relatively few in number and nearly 85% of completed studies do not include PRO. PMID- 17403913 TI - Effect of arylamine acetyltransferase Nat3 gene knockout on N-acetylation in the mouse. AB - Arylamine N-acetyltransferases (NAT) catalyze the biotransformation of many important arylamine drugs and procarcinogens. NAT can either detoxify or activate procarcinogens, complicating the manner in which these enzymes may participate in enhancing or preventing toxic responses to particular agents. Mice possess three NAT isoenzymes: Nat1, Nat2, and Nat3. Whereas Nat1 and Nat2 can efficiently acetylate many arylamines, few substrates appear to be appreciably metabolized by Nat3. We generated a Nat3 knockout mouse strain and used it along with our double Nat1/2(-/-) knockout strain to further investigate the functional role of Nat3. Nat3(-/-) mice showed normal viability and reproductive capacity. Nat3 expression was very low in wild-type animals and completely undetectable in Nat3(-/-) mice. In contrast, greatly elevated expression of Nat3 transcript was observed in Nat1/2(-/-) mice. We used a transcribed marker polymorphism approach to establish that the increased expression of Nat3 in Nat1/2(-/-) mice is a positional artifact of insertion of the phosphoglycerate kinase-neomycin resistance cassette in place of the Nat1/Nat2 gene region and upstream of the intact Nat3 gene, rather than a biological compensatory mechanism. Despite the increase in Nat3 transcript, the N-acetylation of p-aminosalicylate, sulfamethazine, 2 aminofluorene, and 4-aminobiphenyl was undetectable either in vivo or in vitro in Nat1/2(-/-) animals. In parallel, no difference was observed in the in vivo clearance or in vitro metabolism of any of these substrates between wild-type and Nat3(-/-) mice. Thus, Nat3 is unlikely to play a significant role in the N acetylation of arylamines either in wild-type mice or in mice lacking Nat1 and Nat2 activities. PMID- 17403914 TI - Identification of a novel glutathione conjugate of flutamide in incubations with human liver microsomes. AB - Flutamide, a nonsteroidal antiandrogen drug widely used in the treatment of prostate cancer, has been associated with rare incidences of hepatotoxicity in patients. It is believed that bioactivation of flutamide and subsequent covalent binding to cellular proteins is responsible for its toxicity. Current in vitro studies were undertaken to probe the cytochrome P450 (P450)-mediated bioactivation of flutamide and identify the possible reactive species using reduced glutathione (GSH) as a trapping agent. NADPH- and GSH-supplemented human liver microsomal incubations of flutamide gave rise to a novel GSH conjugate where GSH moiety was conjugated to the flutamide molecule via the amide nitrogen, resulting in a sulfenamide. The structure of the conjugate was characterized by liquid chromatography-tandem mass spectrometry and NMR experiments. The conjugate formation was primarily catalyzed by heterologously expressed CYP2C19, CYP1A2, and, to a lesser extent, CYP3A4 and CYP3A5. The mechanism for the formation of this conjugate is unknown; however, a tentative bioactivation mechanism involving a P450-catalyzed abstraction of hydrogen atom from the amide nitrogen of flutamide and the subsequent trapping of the nitrogen-centered radical by GSH or oxidized glutathione (GSSG) was proposed. Interestingly, the same adduct was formed when flutamide was incubated with human liver microsomes in the presence of GSSG and NADPH. This finding suggests that P450-mediated oxidation of flutamide via a nitrogen-centered free radical could be one of the several bioactivation pathways of flutamide. Even though the relationship of the GSH conjugate to flutamide-induced toxicity is unknown, the results have revealed the formation of a novel, hitherto unknown, GSH adduct of flutamide. PMID- 17403915 TI - Biotransformation of 6-methoxy-3-(3',4',5'-trimethoxy-benzoyl)-1H-indole (BPR0L075), a novel antimicrotubule agent, by mouse, rat, dog, and human liver microsomes. AB - 6-Methoxy-3-(3',4',5'-trimethoxy-benzoyl)-1H-indole (BPR0L075) is a novel synthetic indole compound with microtubule binding activity. Incubation of BPR0L075 with mouse, rat, dog, and human liver microsomes in the presence of NADPH resulted in the formation of six metabolites. Liquid chromatography-tandem mass spectrometry and comparison with the synthetic reference standards identified two metabolites (M1 and M5) as the products derived from hydroxylation on the indole moiety of the molecule. M3 was also identified as a product derived from hydroxylation, but the structure of this metabolite was not identified because of the lack of a reference standard. M2, M4, and M6 were identified as the products derived from O-demethylation. M2, 6-desmethyl-BPR0L075, was the major metabolite formed by the liver microsomes of the four species. No qualitative species difference in the metabolism of BPR0L075 was observed. There was quantitative species difference in the metabolism of BPR0L075 among the four species. Whereas mouse and rat liver microsomes metabolized BPR0L075 predominantly via O-demethylation, dog liver microsomes metabolized BPR0L075 by O demethylation and hydroxylation to about the same extent. The rank order of intrinsic clearance rates for the conversion of BPR0L075 to 6-desmethyl-BPR0L075 was mouse > rat > human > dog. Incubation of BPR0L075 with baculovirus-insect cell-expressed human cytochrome P450 (P450) isozymes showed that CYP1A2, 2C9, 2C19, 2D6, 2E1, and 3A4 all catalyzed the O-demethylation and hydroxylation of BPR0L075 but to a different degree. Among the six P450 isozymes tested, CYP1A2 and 2D6 were most active on catalyzing the metabolism of BPR0L075. CYP1A2 catalyzed mainly the formation of M1, M2, and M3. M2 was the predominant metabolite formed by CYP2D6. PMID- 17403916 TI - The disposition of prasugrel, a novel thienopyridine, in humans. AB - Prasugrel, a prodrug, is a novel and potent inhibitor of platelet aggregation in vivo. The metabolism of prasugrel and the elimination and pharmacokinetics of its active metabolite, 2-[1-[2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl]-4-mercapto 3-piperidinylidene]acetic acid (R-138727), three inactive metabolites, and radioactivity were determined in five healthy male subjects after a single 15-mg (100 microCi) p.o. dose of [(14)C]prasugrel. Prasugrel was rapidly absorbed, and maximum plasma concentrations of radioactivity and R-138727 were achieved in 30 min, indicating rapid formation of R-138727. Prasugrel was extensively metabolized in humans, first by hydrolysis to a thiolactone, followed by ring opening to form R-138727, which was further metabolized by S-methylation and conjugation with cysteine. Total radioactivity was higher in plasma than in blood, suggesting limited penetration of prasugrel metabolites into red blood cells. Approximately 70% of the dose was excreted in the urine and 25% in the feces. PMID- 17403917 TI - The phosphoinositide-specific phospholipase C inhibitor U73122 (1-(6-((17beta-3 methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione) spontaneously forms conjugates with common components of cell culture medium. AB - Phosphoinositide-specific phospholipase C (PLC) is a key enzyme in the regulation of Ca(2+) release from inositol 1,4,5-triphosphate-sensitive stores. U73122 (1-(6 ((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione) has been extensively used as a pharmacological inhibitor of PLC to elucidate the importance of this enzyme family in signal transduction pathways. U73122 has an electrophilic maleimide group, which readily reacts with nucleophiles such as thiols and amines. In the current study the conjugation of U73122 to common components of cell culture medium, namely l-glutamine, glutathione, and bovine serum albumin (BSA), was demonstrated. The half-life of U73122 on incubation with phosphate-buffered saline (PBS), Hanks' buffered saline solution (with 2 mM glutamine), optimized basal nutrient medium (MCDB131, without BSA), complete medium, Dulbecco's modified Eagle's medium (with 2 mM l-glutamine) was approximately 150, 60, 32, 30, and 18 min, respectively. However, U73122 was not recoverable from medium supplemented with 0.5% BSA. U73122 underwent hydrolysis of the maleimide group when incubated with PBS. Glutamine conjugates of U73122 were identified in cell culture medium. Furthermore, the inhibition of epidermal growth factor-stimulated Ca(2+) release in a human epidermoid carcinoma cell line (A431) by U73122 was substantially reduced by the presence of BSA in a time dependent manner. In complex cellular assays, the availability of U73122 to inhibit PLC may be limited by its chemical reactivity and lead to the misinterpretation of results in pharmacological assays. PMID- 17403918 TI - Erythropoietin modulation of podocalyxin and a proposed erythroblast niche. AB - Epo's erythropoietic capacity is ascribed largely to its antiapoptotic actions. In part via gene profiling of bone marrow erythroblasts, Epo is now shown to selectively down-modulate the adhesion/migration factors chemokine receptor-4 (Cxcr4) and integrin alpha-4 (Itga4) and to up-modulate growth differentiation factor-3 (Gdf3), oncostatin-M (OncoM), and podocalyxin like-1 (PODXL). For PODXL, Epo dose-dependent expression of this CD34-related sialomucin was discovered in Kit(+)CD71(high) proerythroblasts and was sustained at subsequent Kit( )CD71(high) and Ter119(+) stages. In vivo, Epo markedly induced PODXL expression in these progenitors and in marrow-resident reticulocytes. This was further associated with a rapid release of PODXL(+) reticulocytes to blood. As studied in erythroblasts expressing minimal Epo receptor (EpoR) alleles, efficient PODXL induction proved dependence on an EpoR-PY343 Stat5 binding site. Moreover, in mice expressing an EpoR-HM F343 allele, compromised Epo-induced PODXL expression correlated with abnormal anucleated red cell representation in marrow. By modulating this select set of cell-surface adhesion molecules and chemokines, Epo is proposed to mobilize erythroblasts from a hypothesized stromal niche and possibly promote reticulocyte egress to blood. PMID- 17403919 TI - Enhanced ability of dendritic cells to stimulate innate and adaptive immunity on short-term incubation with zoledronic acid. AB - Vgamma9/Vdelta2 (gammadelta) T cells play a major role in innate immunity against microbes, stressed, and tumor cells. They represent less than 5% of peripheral blood lymphocytes but can be activated and expanded in vitro by aminobisphosphonates (ABP)-treated monocytes. The aim of this work was to determine whether ABP-treated dendritic cells (DCs) can also activate gammadelta T cells and regulate immune responses mediated by conventional alphabeta T cells. Highly purified immature (iDC) and mature DC (mDC) were generated from peripheral blood monocytes of healthy donors and incubated with zoledronic acid (Zol) for 24 hours. Zol-treated iDC and mDC retained their immunostimulatory properties and induced the vigorous expansion of central memory and effector memory gammadelta T cells. gammadelta T cells displayed antitumor activity and appropriate cell surface antigens to target secondary lymphoid organs and exert costimulatory activity. Antigen-specific MHC-restricted immune responses, mediated by conventional alphabeta T cells, were improved by the concurrent gammadelta T-cell activation. In conclusion, large numbers of gammadelta T cells with effector and costimulatory activities are rapidly generated by Zol-treated iDC/mDC. This strategy is worthy of further investigation to improve adoptive cell therapy and vaccine interventions against tumors and infections. PMID- 17403920 TI - Abstracts from the ACOG (American College of Obstetricians and Gynecologists) 55th Annual Clinical Meeting. May 5-9, 2007. San Diego, California, USA. PMID- 17403925 TI - Pseudomonas aeruginosa exploits a PIP3-dependent pathway to transform apical into basolateral membrane. AB - Pseudomonas aeruginosa, an important human pathogen, preferentially binds and enters injured cells from the basolateral (BL) surface. We previously demonstrated that activation of phosphatidylinositol 3-kinase (PI3K) and Akt are necessary and sufficient for P. aeruginosa entry from the apical (AP) surface and that AP addition of phosphatidylinositol 3,4,5-trisphosphate (PIP3) is sufficient to convert AP into BL membrane (Kierbel, A., A. Gassama-Diagne, K. Mostov, and J.N. Engel. 2005. Mol. Biol. Cell. 16:2577-2585; Gassama-Diagne, A., W. Yu, M. ter Beest, F. Martin-Belmonte, A. Kierbel, J. Engel, and K. Mostov. 2006. Nat. Cell Biol. 8:963-970). We now show that P. aeruginosa subverts this pathway to gain entry from the AP surface. In polarized monolayers, P. aeruginosa binds near cell-cell junctions without compromising them where it activates and recruits PI3K to the AP surface. Membrane protrusions enriched for PIP3 and actin accumulate at the AP surface at the site of bacterial binding. These protrusions lack AP membrane markers and are comprised of BL membrane constituents, which are trafficked there by transcytosis. The end result is that this bacterium transforms AP into BL membrane, creating a local microenvironment that facilitates its colonization and entry into the mucosal barrier. PMID- 17403926 TI - The role of karyopherins in the regulated sumoylation of septins. AB - In the yeast Saccharomyces cerevisiae, several components of the septin ring are sumoylated during anaphase and then abruptly desumoylated at cytokinesis. We show that septin sumoylation is controlled by the interactions of two enzymes of the sumoylation pathway, Siz1p and Ulp1p, with the nuclear transport machinery. The E3 ligase Siz1p is imported into the nucleus by the karyopherin Kap95p during interphase. In M phase, Siz1p is exported from the nucleus by the karyopherin Kap142p/Msn5p and subsequently targeted to the septin ring, where it participates in septin sumoylation. We also show that the accumulation of sumoylated septins during mitosis is dependent on the interactions of the SUMO isopeptidase Ulp1p with Kap121p and Kap95p-Kap60p and the nuclear pore complex (NPC). In addition to sequestering Ulp1 at the NPC, Kap121p is required for targeting Ulp1p to the septin ring during mitosis. We present a model in which Ulp1p is maintained at the NPC during interphase and transiently interacts with the septin ring during mitosis. PMID- 17403927 TI - Myosin VI is required for sorting of AP-1B-dependent cargo to the basolateral domain in polarized MDCK cells. AB - In polarized epithelial cells, newly synthesized membrane proteins are delivered on specific pathways to either the apical or basolateral domains, depending on the sorting motifs present in these proteins. Because myosin VI has been shown to facilitate secretory traffic in nonpolarized cells, we investigated its role in biosynthetic trafficking pathways in polarized MDCK cells. We observed that a specific splice isoform of myosin VI with no insert in the tail domain is required for the polarized transport of tyrosine motif containing basolateral membrane proteins. Sorting of other basolateral or apical cargo, however, does not involve myosin VI. Site-directed mutagenesis indicates that a functional complex consisting of myosin VI, optineurin, and probably the GTPase Rab8 plays a role in the basolateral delivery of membrane proteins, whose sorting is mediated by the clathrin adaptor protein complex (AP) AP-1B. Our results suggest that myosin VI is a crucial component in the AP-1B-dependent biosynthetic sorting pathway to the basolateral surface in polarized epithelial cells. PMID- 17403928 TI - Phosphorylation and regulation of a G protein-coupled receptor by protein kinase CK2. AB - We demonstrate a role for protein kinase casein kinase 2 (CK2) in the phosphorylation and regulation of the M3-muscarinic receptor in transfected cells and cerebellar granule neurons. On agonist occupation, specific subsets of receptor phosphoacceptor sites (which include the SASSDEED motif in the third intracellular loop) are phosphorylated by CK2. Receptor phosphorylation mediated by CK2 specifically regulates receptor coupling to the Jun-kinase pathway. Importantly, other phosphorylation-dependent receptor processes are regulated by kinases distinct from CK2. We conclude that G protein-coupled receptors (GPCRs) can be phosphorylated in an agonist-dependent fashion by protein kinases from a diverse range of kinase families, not just the GPCR kinases, and that receptor phosphorylation by a defined kinase determines a specific signalling outcome. Furthermore, we demonstrate that the M3-muscarinic receptor can be differentially phosphorylated in different cell types, indicating that phosphorylation is a flexible regulatory process where the sites that are phosphorylated, and hence the signalling outcome, are dependent on the cell type in which the receptor is expressed. PMID- 17403929 TI - Dolichol-linked oligosaccharide selection by the oligosaccharyltransferase in protist and fungal organisms. AB - The dolichol-linked oligosaccharide Glc3Man9GlcNAc2-PP-Dol is the in vivo donor substrate synthesized by most eukaryotes for asparagine-linked glycosylation. However, many protist organisms assemble dolichol-linked oligosaccharides that lack glucose residues. We have compared donor substrate utilization by the oligosaccharyltransferase (OST) from Trypanosoma cruzi, Entamoeba histolytica, Trichomonas vaginalis, Cryptococcus neoformans, and Saccharomyces cerevisiae using structurally homogeneous dolichol-linked oligosaccharides as well as a heterogeneous dolichol-linked oligosaccharide library. Our results demonstrate that the OST from diverse organisms utilizes the in vivo oligo saccharide donor in preference to certain larger and/or smaller oligosaccharide donors. Steady state enzyme kinetic experiments reveal that the binding affinity of the tripeptide acceptor for the protist OST complex is influenced by the structure of the oligosaccharide donor. This rudimentary donor substrate selection mechanism has been refined in fungi and vertebrate organisms by the addition of a second, regulatory dolichol-linked oligosaccharide binding site, the presence of which correlates with acquisition of the SWP1/ribophorin II subunit of the OST complex. PMID- 17403930 TI - 12R-lipoxygenase deficiency disrupts epidermal barrier function. AB - 12R-lipoxygenase (12R-LOX) and the epidermal LOX-3 (eLOX-3) constitute a novel LOX pathway involved in terminal differentiation in skin. This view is supported by recent studies showing that inactivating mutations in 12R-LOX and eLOX-3 are linked to the development of autosomal recessive congenital ichthyosis. We show that 12R-LOX deficiency in mice results in a severe impairment of skin barrier function. Loss of barrier function occurs without alterations in proliferation and stratified organization of the keratinocytes, but is associated with ultrastructural anomalies in the upper granular layer, suggesting perturbance of the assembly/extrusion of lamellar bodies. Cornified envelopes from skin of 12R LOX-deficient mice show increased fragility. Lipid analysis demonstrates a disordered composition of ceramides, in particular a decrease of ester-bound ceramide species. Moreover, processing of profilaggrin to monomeric filaggrin is impaired. This study indicates that the 12R-LOX-eLOX-3 pathway plays a key role in the process of epidermal barrier acquisition by affecting lipid metabolism, as well as protein processing. PMID- 17403931 TI - A role for a novel centrosome cycle in asymmetric cell division. AB - Tissue stem cells play a key role in tissue maintenance. Drosophila melanogaster central brain neuroblasts are excellent models for stem cell asymmetric division. Earlier work showed that their mitotic spindle orientation is established before spindle formation. We investigated the mechanism by which this occurs, revealing a novel centrosome cycle. In interphase, the two centrioles separate, but only one is active, retaining pericentriolar material and forming a "dominant centrosome." This centrosome acts as a microtubule organizing center (MTOC) and remains stationary, forming one pole of the future spindle. The second centriole is inactive and moves to the opposite side of the cell before being activated as a centrosome/MTOC. This is accompanied by asymmetric localization of Polo kinase, a key centrosome regulator. Disruption of centrosomes disrupts the high fidelity of asymmetric division. We propose a two-step mechanism to ensure faithful spindle positioning: the novel centrosome cycle produces a single interphase MTOC, coarsely aligning the spindle, and spindle-cortex interactions refine this alignment. PMID- 17403932 TI - c-Met is essential for wound healing in the skin. AB - Wound healing of the skin is a crucial regenerative process in adult mammals. We examined wound healing in conditional mutant mice, in which the c-Met gene that encodes the receptor of hepatocyte growth factor/scatter factor was mutated in the epidermis by cre recombinase. c-Met-deficient keratinocytes were unable to contribute to the reepithelialization of skin wounds. In conditional c-Met mutant mice, wound closure was slightly attenuated, but occurred exclusively by a few (5%) keratinocytes that had escaped recombination. This demonstrates that the wound process selected and amplified residual cells that express a functional c Met receptor. We also cultured primary keratinocytes from the skin of conditional c-Met mutant mice and examined them in scratch wound assays. Again, closure of scratch wounds occurred by the few remaining c-Met-positive cells. Our data show that c-Met signaling not only controls cell growth and migration during embryogenesis but is also essential for the generation of the hyperproliferative epithelium in skin wounds, and thus for a fundamental regenerative process in the adult. PMID- 17403933 TI - A distal effect of microsomal triglyceride transfer protein deficiency on the lysosomal recycling of CD1d. AB - Microsomal triglyceride transfer protein (MTP) is an endoplasmic reticulum (ER) resident lipid transfer protein involved in the biosynthesis and lipid loading of apolipoprotein B. MTP was recently suggested to directly regulate the biosynthesis of the MHC I-like, lipid antigen presenting molecule CD1d, based on coprecipitation experiments and lipid loading assays. However, we found that the major impact of MTP deficiency occurred distal to the ER and Golgi compartments. Thus, although the rates of CD1d biosynthesis, glycosylation maturation, and internalization from the cell surface were preserved, the late but essential stage of recycling from lysosome to plasma membrane was profoundly impaired. Likewise, functional experiments indicated defects of CD1d-mediated lipid presentation in the lysosome but not in the secretory pathway. These intriguing findings suggest a novel, unexpected role of MTP at a late stage of CD1d trafficking in the lysosomal compartment. PMID- 17403934 TI - Spontaneous tumor rejection by cbl-b-deficient CD8+ T cells. AB - The concept of tumor surveillance implies that specific and nonspecific components of the immune system eliminate tumors in the early phase of malignancy. Understanding the biochemical mechanisms of tumor immunosurveillance is of paramount significance because it might allow one to specifically modulate spontaneous antitumor activity. We report that inactivation of the E3 ligase Casitas B cell lymphoma-b (Cbl-b) confers spontaneous in vivo rejection of tumor cells that express human papilloma virus antigens. Moreover, cbl-b(-/-) mice develop significantly fewer ultraviolet B (UVB)-induced skin malignancies and reject UVB-induced skin tumors. CD8(+) T cells were identified as key players in the spontaneous tumor rejection response. Loss of Cbl-b not only enhances antitumor reactivity of CD8(+) T cells but also occurs in the absence of CD4(+) T cells. Mechanistically, cbl-b(-/-) CD8(+) T cells are resistant to T regulatory cell-mediated suppression and exhibit enhanced activation and rapid tumor infiltration. Importantly, therapeutic transfer of naive cbl-b(-/-) CD8(+) T cells is sufficient to mediate rejection of established tumors. Even up to 1 yr after the first encounter with the tumor cells, cbl-b(-/-) mice carry an "anticancer memory." These data identify Cbl-b as a key signaling molecule that controls spontaneous antitumor activity of cytotoxic T cells in different cancer models. Inhibition of Cbl-b is a novel approach to stimulate long-lasting immunity against cancer. PMID- 17403935 TI - Repression of BCL-6 is required for the formation of human memory B cells in vitro. AB - Memory B cells provide rapid protection to previously encountered antigens; however, how these cells develop from germinal center B cells is not well understood. A previously described in vitro culture system using human tonsillar germinal center B cells was used to study the transcriptional changes that occur during differentiation of human memory B cells. Kinetic studies monitoring the expression levels of several known late B cell transcription factors revealed that BCL-6 is not expressed in memory B cells generated in vitro, and gene expression profiling studies confirmed that BCL-6 is not expressed in these memory B cells. Furthermore, ectopic expression of BCL-6 in human B cell cultures resulted in formation of fewer memory B cells. In addition, the expression profile of in vitro memory B cells showed a unique pattern that includes expression of genes encoding multiple costimulatory molecules and cytokine receptors, antiapoptotic proteins, T cell chemokines, and transcription factors. These studies establish new molecular criteria for defining the memory B cell stage in human B cells. PMID- 17403936 TI - Importance of group X-secreted phospholipase A2 in allergen-induced airway inflammation and remodeling in a mouse asthma model. AB - Arachidonic acid metabolites, the eicosanoids, are key mediators of allergen induced airway inflammation and remodeling in asthma. The availability of free arachidonate in cells for subsequent eicosanoid biosynthesis is controlled by phospholipase A(2)s (PLA(2)s), most notably cytosolic PLA(2)-alpha. 10 secreted PLA(2)s (sPLA(2)s) have also been identified, but their function in eicosanoid generation is poorly understood. We investigated the role of group X sPLA(2) (sPLA(2)-X), the sPLA(2) with the highest in vitro cellular phospholipolysis activity, in acute and chronic mouse asthma models in vivo. The lungs of sPLA(2) X(-/-) mice, compared with those of sPLA(2)-X(+/+) littermates, had significant reduction in ovalbumin-induced infiltration by CD4(+) and CD8(+) T cells and eosinophils, goblet cell metaplasia, smooth muscle cell layer thickening, subepithelial fibrosis, and levels of T helper type 2 cell cytokines and eicosanoids. These data direct attention to sPLA(2)-X as a novel therapeutic target for asthma. PMID- 17403937 TI - Magnetically driven plasmid DNA delivery with biodegradable polymeric nanoparticles. AB - Targeting gene therapy remains a challenge. The use of magnetic force to achieve this was investigated in the present study. It was hypothesized that nanoparticles with both controllable particle size and magnetic properties would enable magnetically driven gene delivery. We investigated this hypothesis by creating a family of novel biodegradable polymeric superparamagnetic nanoparticle (MNP) formulations. Polylactide MNP were formulated using a modified emulsification-solvent evaporation methodology with both the incorporation of oleate-coated iron oxide and a polyethylenimine (PEI) oleate ion-pair surface modification for DNA binding. MNP size could be controlled by varying the proportion of the tetrahydrofuran cosolvent. Magnetically driven MNP-mediated gene transfer was studied using a green fluorescent protein reporter plasmid in cultured arterial smooth muscle cells and endothelial cells. MNP-DNA internalization and trafficking were examined by confocal microscopy. Cell growth inhibition after MNP-mediated adiponectin plasmid transfection was studied as an example of a therapeutic end point. MNP-DNA complexes protected DNA from degradation and efficiently transfected quiescent cells under both low and high serum conditions after a 15 min exposure to a magnetic field (500 G). There was negligible transfection with MNP in the absence of a magnetic field. Larger sized MNP (375 nm diameter) exhibited higher transfection rates compared with 185 nm- and 240 nm-sized MNP. Internalized larger sized MNP escaped lysosomal localization and released DNA in the perinuclear zone. Adiponectin plasmid DNA delivery using MNP resulted in a dose-dependent growth inhibition of cultured arterial smooth muscle cells. It is concluded that magnetically driven plasmid DNA delivery can be achieved using biodegradable MNP containing oleate-coated magnetite and surface modified with PEI oleate ion-pair complexes that enable DNA binding. PMID- 17403938 TI - Maintaining precursor pools for mitochondrial DNA replication. AB - Among the human diseases that result from abnormalities in mitochondrial genome stability or maintenance are several that result from mutations affecting enzymes of deoxyribonucleoside triphosphate (dNTP) metabolism. In addition, it is evident that the toxicity of antiviral nucleoside analogs is determined in part by the extent to which their intracellular conversion to dNTP analogs occurs within the mitochondrion. Finally, recent work from this laboratory has shown considerable variation among different mammalian tissues with respect to mitochondrial dNTP pool sizes and has suggested that natural asymmetries in mitochondrial dNTP concentrations may contribute to the high rates at which the mitochondrial genome undergoes mutation. These factors suggest that much more information is needed about maintenance and regulation of dNTP pools within mammalian mitochondria. This review summarizes our current understanding and suggests directions for future research. PMID- 17403939 TI - Single-dose intracerebroventricular administration of galactocerebrosidase improves survival in a mouse model of globoid cell leukodystrophy. AB - Globoid cell leukodystrophy (GLD), also known as Krabbe disease, is a devastating, degenerative neurological disorder. It is inherited as an autosomal recessive trait caused by loss-of-function mutations in the galactocerebrosidase (GALC) gene. Previously, we have shown that peripheral injection of recombinant GALC, administered every other day, results in a substantial improvement in early clinical phenotype in the twitcher mouse model of GLD. While we did detect active enzyme in the brain following peripheral administration, most of the administered enzyme was localized to the periphery. Given the substantial central nervous system (CNS) involvement in this disease, we were interested in determining whether or not a single-dose administration of the recombinant enzyme directly to the CNS, which could potentially be achieved clinically, would result in any substantial improvement. Following intracerebroventricular (i.c.v.) administration of GALC we noted a significant, 16.5%, reduction in the GALC substrate psychosine, the abnormal accumulation of which is believed to play a pivotal role in the CNS pathology observed in this disease. Moreover, recombinant GALC was found not only in periventricular regions but also at sites distant to the injection such as the cerebral cortex and cerebellum. Most importantly, animals receiving a single i.c.v. dose of the enzyme at postnatal day 20 survived up to 51 days, which compares favorably to the control twitcher animals, which normally only live to postnatal day 40/42. These results indicate that even a single i.c.v. administration of the recombinant enzyme can have significant clinical impact and suggests that other lysosomal storage disorders with significant CNS involvement may similarly benefit. PMID- 17403940 TI - Transplantation of embryonic stem cells improves nerve repair and functional recovery after severe sciatic nerve axotomy in rats. PMID- 17403941 TI - Multiply infected vectors. PMID- 17403942 TI - Measuring species richness based on microbial community fingerprints: the emperor has no clothes. PMID- 17403943 TI - Predictions under isothermal and dynamically changing conditions. PMID- 17403944 TI - The use of elearning in medical education. PMID- 17403945 TI - The use of elearning in medical education: a review of the current situation. AB - Computers are increasingly used in medical education. Electronic learning (elearning) is moving from textbooks in electronic format (that are increasingly enhanced by the use of multimedia adjuncts) to a truly interactive medium that can be delivered to meet the educational needs of students and postgraduate learners. Computer technology can present reliable, reusable content in a format that is convenient to the learner. It can be used to transcend geographical boundaries and time zones. It is a valuable tool to add to the medical teacher's toolkit, but like all tools it must be used appropriately. This article endeavours to review the current "state of the art2 in use of elearning and its role in medical education alongside non-electronic methods-a combination that is currently referred to as "blended" learning. PMID- 17403946 TI - Management of paraproteinaemia. AB - A paraprotein is a monoclonal immunoglobulin or light chain present in the blood or urine; it is produced by a clonal population of mature B cells, most commonly plasma cells. In individuals aged >50 years the incidence of a paraprotein is 3.2%. Plasma cell disorders can be considered as a spectrum of conditions ranging from monoclonal gammopathy of undetermined significance (MGUS), through asymptomatic, to symptomatic myeloma. MGUS is defined by a low level of paraprotein <30 g/l, bone marrow plasma cells <10% and the absence of myeloma related organ or tissue damage (predominantly renal, skeletal or bone marrow impairment.) MGUS requires no therapy and the overall risk of progression to myeloma is 1% per year. Myeloma remains incurable with a median survival of 3-4 years; autologous stem cell transplant can prolong survival, if appropriate. Thalidomide in combination with dexamethasone has an emerging role in the treatment of myeloma. PMID- 17403947 TI - Islet cell transplantation. AB - People with type 1 diabetes have normal exocrine pancreatic function, making islet cell rather than whole organ transplantation an attractive option. Achieving insulin independence in type 1 diabetes was the perceived goal of islet cell transplantation. The success of the Edmonton group in achieving this in a selected group of type 1 patients has led to renewed optimism that this treatment could eventually replace whole organ pancreas transplantation. However the long term results of this treatment indicate that insulin independence is lost with time in a significant proportion of patients, although they may retain glycaemic stability. In this context, the indications for islet cell transplantation, which have evolved over the last 5 years, indicate that the patients who benefit most are those who experience severe hypoglycaemic reactions despite optimal insulin therapy. This review will summarise the history of islet cell transplantation, islet isolation techniques, the transplant procedure, immunosuppressive therapy, indications for islet cell transplantation, current clinical trials, the early UK islet cell transplant experience using the Edmonton protocol, and some of the challenges that lie ahead. PMID- 17403948 TI - Vitamin D in childhood and adolescence. AB - It is well-established that prolonged and severe vitamin D deficiency leads to rickets in children and osteomalacia in adults. More marginal vitamin D deficiency is likely to be a significant contributing factor to osteoporosis risk. However, recent emerging data from studies of adults suggest that low vitamin D status (serum 25-hydroxyvitamin D levels <50 nmol/l) may be contributing to the development of various chronic diseases, including cardiovascular disease, hypertension, diabetes mellitus, some inflammatory and autoimmune diseases, and certain cancers. Adequacy of vitamin D status in children and adolescents has been the focus of a number of recent investigations, and these studies have shown a high prevalence of low vitamin D status during the winter (especially in adolescents), with lower prevalence during the summer. Therefore, consideration of potential corrective strategies to allow children and adolescents to maintain adequate vitamin D status throughout the year, even in the absence of adequate summer sun exposure, is warranted. PMID- 17403949 TI - Classical diseases revisited: transient global amnesia. AB - Transient global amnesia usually affects patients between the ages of 40 and 80. Patients with this condition are often described--wrongly--as being confused. It presents classically with an abrupt onset of severe anterograde amnesia. It is usually accompanied by repetitive questioning. The patient does not have any focal neurological symptoms. Patients remain alert, attentive, and cognition is not impaired. However, they are disoriented to time and place. Attacks usually last for 1-8 h but should be less than 24 h. It is possible that it may result from different mechanisms such as venous congestion with valsalva-like activities before symptom onset, arterial thromboembolic ischaemia and vasoconstriction due to hyperventilation. Diagnosis may be made safely in the presence of a characteristic collateral history. No specific treatment is indicated for a typical episode. PMID- 17403950 TI - Recent development of ischaemic heart disease in sex difference. AB - Despite a dramatic decline in mortality over the past years, coronary heart disease is the leading cause of death and disability in the world. At the same time, with the great improvement of medical science, there is a growing population of postmyocardial infarction, postrevascularisation and heart failure survivors. Furthermore, there are rising rates of cigarette smoking, obesity, hypertension and the metabolic syndrome in the world. All the above contribute to the rising incidence rates of ischaemic heart disease (IHD) among women and men. This review highlights sex-specific issues in IHD presentation, evaluation and outcomes, with several new results published from the Women's Ischemia Syndrome Evaluation study. New evidence on traditional and novel risk markers as well as sex-specific differences in symptoms and diagnostic approaches have also been discussed. PMID- 17403951 TI - DNA sequence variations of metalloproteinases: their role in asthma and COPD. AB - Asthma and chronic obstructive pulmonary disease (COPD) are complex genetic diseases that cause considerable morbidity and mortality worldwide. Genetic variability interacting with environmental and ethnic factors is presumed to cause tobacco smoke susceptibility and to influence asthma severity. A disintegrin and metalloproteinase 33 (ADAM33) and matrix metalloproteinase-9 (MMP9) appear to have important roles in asthma and COPD pathogenesis. ADAM33 and MMP9 genetic alterations could possibly contribute to the establishment and progression of these multifactorial diseases, although their association with the clinical phenotypes has not yet been elucidated. However, the occurrence of these alterations does not always result in clear disease, implying that either they are an epiphenomenon or they are in proximity to the true causative alteration. This review summarises the most recent literature dealing with the genetic variations of metalloproteinases and outlines their potential pathogenetic outcome. PMID- 17403952 TI - Immune-mediated inflammatory diseases (IMIDs) and biologic therapy: a medical revolution. AB - Targeted biologic therapies have revolutionised treatment of immune-mediated inflammatory diseases (IMIDs) due to their efficacy, speed of onset and tolerability. The discovery that clinically unrelated conditions, such as rheumatoid arthritis and Crohn's disease, share similar immune dysregulation has led to a shift in the management of IMIDs from one of organ-based symptom relief to mechanism-based treatment. The fact that anticytokine therapy has been effective in treating multiple orphan inflammatory conditions confirms the IMID paradigm. In this review we examine the biologic agents currently licensed for use in the US and Europe: infliximab, etanercept, adalimumab, rituximab, abatacept, anakinra, alefacept and efalizumab. We also discuss the rationale behind the management of IMIDs using rheumatoid arthritis, Crohn's disease, psoriasis and psoriatic arthritis as examples. For the medical profession, IMID represents a breakthrough in the way pathology is classified. In this burgeoning era of biologic therapy the prospect of complete disease remission is conceivable. PMID- 17403953 TI - Tako-tsubo syndrome on the rise: a review of the current literature. AB - Tako-tsubo syndrome (TTS) or stress-related acute reversible ventricular apical dysfunction is an emerging but seemingly under-recognised cardiomyopathy mimicking acute ST elevation myocardial infarction (STEMI) without concomitant epicardial coronary artery disease. Severe emotional stress is the most common trigger for this syndrome in the published series, but it can also be precipitated by severe intercurrent medical illness. Precise epidemiological data are not yet available, although TTS most commonly affects elderly women. The exact cause of this syndrome is undetermined, but proposed mechanisms include epicardial coronary artery vasospasm, impaired multivessel coronary microcirculation, calcium overload with direct myocyte damage and disrupted fatty acid metabolism with prolonged myocardial stunning. The time course of electrocardiographic changes is very similar to that of an acute STEMI due to an acute occlusion of the left anterior descending coronary artery. The left ventricular dysfunction typically displays an akinetic apical half of the left or both ventricles with hyperkinetic basal segments, although a variant with apical sparing has also been described recently. The ventricular dysfunction usually resolves within weeks and carries a generally favourable prognosis. PMID- 17403954 TI - Resuscitation competencies in paediatric specialist registrars. AB - BACKGROUND: Despite the increased availability of resuscitation courses and guidelines requiring optimal training for acute situations, little is known on the actual qualification of house staff with in-hospital on-call duties for critically ill newborns and children. OBJECTIVE: To assess (1) the characteristics of education and training; (2) the level of experience; and (3) factors that may hamper optimal performance of paediatric specialist registrars caring for acute critically ill newborns and children. METHODS: A structured questionnaire was completed by a national cohort of all paediatric specialist registrars in their fourth year (ie pre-final) of training. RESULTS: Important shortcomings in training and assessment of actual qualifications of resuscitation competencies were identified in paediatric specialist registrars. In 17 of 57 (30%) specialist registrars, competencies in acute care had never been assessed or reconfirmed when starting on-call duties while in the others, 40 (70%), substantial heterogeneity was found regarding the type of assessment of qualifications for on-call duties. In acute situations, occasionally untrained and unsupervised resuscitation procedures were performed. Individual responsibility was the most important stressor that may hamper optimal performance. Despite these findings, adequate reported levels of self-confidence were found; self confidence was higher in newborns as compared to children (7.8 and 7.0 respectively on scale 1-10, p<0.05). CONCLUSIONS: Successful completion of a resuscitation course does not ensure adequate qualifications by on-call residents, unless regular refresher sessions are provided. Teaching hospitals should establish and implement uniform guidelines for training and assessment of competencies regarding acute care for critically ill children and newborns. PMID- 17403955 TI - Orthopaedic surgery core curriculum: the spine. AB - OBJECTIVE: To develop a core curriculum for orthopaedic surgery and to conduct a national survey to assess the importance of 281 items in the curriculum. Attention was focused specifically on 24 items pertaining to the curriculum that are pertinent to the spine. STUDY DESIGN: A cross-sectional survey of a random sample of orthopaedic surgeons whose primary affiliation was non-academic, representing the provinces and territories of Canada METHODS: A questionnaire containing 281 items was developed. A random group of 131 (out of 156) orthopaedic surgeons whose primary affiliation is non-academic completed the questionnaire. The data were analysed quantitatively using average mean scores, histograms, the modified Hotelling's T2 test and the Benjimini-Hochberg procedure. RESULTS: 131 of 156 (84%) orthopaedic surgeons participated, in this study. 14 of 24 items were ranked at no less than 3 out of 4 thus suggesting that 58% of the items are important or probably important to know by the end of residency (SD< or =0.07). Residents need to learn the diagnosis and principles of managing patients with common conditions of the spine. CONCLUSIONS: The study shows, with reliable statistical evidence, that orthopaedic residents are no longer expected to be able to perform spinal fusions with proficiency on completion of residency. Is the exposure to surgical spine problems and the ability to be comfortable with operating expectations specific to the fellowship level? If so, the focus during residency or increasing accredited spine fellowships needs to be addressed to ensure that enough spine surgeons are educated to meet the future healthcare demands projected for Canada. PMID- 17403956 TI - Clinical presentation and incidence of complications in patients with coeliac disease diagnosed by relative screening. AB - BACKGROUND: There is an increased prevalence of coeliac disease (CD) among relatives of those with the disease. AIMS: To compare the clinical features in patients with CD detected via family screening with those in patients diagnosed routinely. METHODS: Information on screening was provided to relatives of patients. Those who wished to be screened were tested for endomysial and/or tissue transglutaminase antibodies. Duodenal biopsy was performed in those with positive antibodies. The clinical details of the relative screening group were compared with those of 105 patients diagnosed routinely. RESULTS: 183 relatives underwent screening, of whom 32 had positive serology, 24 had histology diagnostic of CD, six had normal biopsies and two declined duodenal biopsy. Patients in the relative screening group were younger with a median age of 33 years (range 17-72 years) compared to the routine group which had a median age of 54 years (range 25-88 years). In the relative screening group, there was a male preponderance (M:F ratio 16:8), anaemia at presentation was significantly less common (13% v 58%; p<0.001) and osteoporosis was less frequent (9% v 22%; p<0.244) compared with the routine group. 65% of the relative screening group had gastrointestinal symptoms or anaemia at diagnosis. CONCLUSIONS: Patients detected by family screening are younger with a male preponderance, but fewer had anaemia and osteoporosis. PMID- 17403957 TI - Cardiovascular disease, hypertension, dyslipidaemia and obesity in patients with hypothalamic-pituitary disease. AB - OBJECTIVE: Adults with hypothalamic-pituitary disease have increased morbidity and mortality from cardiovascular disease (CVD). Therefore, the prevalence of CVD and adequacy of treatment of cardiovascular risk factors (according to current treatment guidelines) was studied in a large group of patients with hypothalamic pituitary disease. STUDY DESIGN: In 2005, 152 consecutive adult patients with hypothalamic-pituitary disease attending our neuro-endocrine centre were clinically examined and blood pressure (BP), lipid profile, type 2 diabetes mellitus, body composition and smoking status were assessed. RESULTS: Of the 152 patients, 36.8% had treated hypertension and 28.2% had treated dyslipidaemia. Many of these patients had inadequate BP control (BP >140/85 mm Hg, 44.6%) and undesirable lipid levels (total cholesterol >4.0 mmol/l, 69%). Also, many of the untreated patients had BP and lipid levels which should have been considered for treatment (26 patients (27%) and 83 patients (76%), respectively). Smoking was admitted in 18% of patients. Central adiposity was present in 86% and obesity (body mass index > or =30) was present in 50%. CONCLUSIONS: Cardiovascular risk factors are highly prevalent and often inadequately treated in adult patients with hypothalamic-pituitary disease. Aggressive treatment of these factors is essential to reduce mortality and morbidity from CVD in these patients. PMID- 17403958 TI - In what ways can human skeletal remains be used to understand health and disease from the past? AB - Disease and illness in previous generations can be investigated using palaeopathology. This article describes the commonly used techniques in palaeopathology, includes examples of how such techniques are able to formulate data on a variety of health issues that occurred in the past and suggests how these data can be relevant today. PMID- 17403959 TI - A good death certificate: improved performance by simple educational measures. AB - BACKGROUND AND AIMS: The initial aim of this audit was to determine whether information on death certificates is correct and all legal requirements are met. As shortcomings were found, educational measures were undertaken and the effect of those was measured by a re-audit. METHOD: All death certificates issued during a 4-month period within the elderly care department of a district general hospital were retrospectively audited. A re-audit was performed later the same year over a 3-month period. RESULTS: 19 (13.6%) of 140 certificates issued during the initial 4-month period could not be shown to meet the statutory criteria, as no evidence was found that these patients were attended by the issuing medical officer. Minor errors and omissions were found in 58.6% of certificates. Following education about these problems, there was a significant improvement in death certification. Only 2 (2.4%) of 85 certificates issued in the re-audit period did not meet the statutory criteria (p = 0.01) and minor errors and omissions occurred in 20%. CONCLUSION: The incidence of unsatisfactory death certificates within a hospital setting is high. Increased education and better documentation leads to improvements in accuracy and legitimacy. PMID- 17403961 TI - Longitudinal course of pediatric bipolar disorder. PMID- 17403962 TI - What have we learned about the neurobiology of major depression? PMID- 17403963 TI - Can't get enough of that dopamine. PMID- 17403964 TI - Inroads to mechanisms of disease in child psychiatric disorders. PMID- 17403965 TI - Furor therapeuticus: Benjamin Rush and the Philadelphia yellow fever epidemic of 1793. PMID- 17403966 TI - Fragile X syndrome: molecular mechanisms of cognitive dysfunction. PMID- 17403967 TI - Psychiatric disorders: a conceptual taxonomy. AB - This article summarizes six conceptual dimensions that underlie common assumptions about what counts as an adequate category of psychiatric disorder. These dimensions are 1) causalism-descriptivism, 2) essentialism-nominalism, 3) objectivism-evaluativism, 4) internalism-externalism, 5) entities-agents, and 6) categories-continua. Four different versions of the medical model are described and compared with respect to these dimensions. The medical models vary in several ways, but all can be considered "essentialistic." As a counter to the essentialist homogeneity among the medical models, two nominalist analyses of psychiatric classification are reviewed. In order to fill out the space defined by the conceptual dimensions, two alternatives to medical model approaches are also described. After making some suggestions about where DSM-V might best be aligned with respect to the conceptual dimensions, the authors review the distinction between empirical and nonempirical aspects of classification--and argue that nonempirical aspects of classification are legitimate and necessary. PMID- 17403968 TI - Lifting the veil on trichotillomania. PMID- 17403969 TI - A genetic etiology of pervasive developmental disorder guides treatment. PMID- 17403970 TI - The Lunatic Asylum on Blackwell's Island and the New York Press. PMID- 17403971 TI - Twelve-month outcome of adolescents with bipolar disorder following first hospitalization for a manic or mixed episode. AB - OBJECTIVE: Although adolescent-onset bipolar disorder is associated with significant morbidity, there have been few prospective outcome studies of this population. The aim of this study was to examine the 12-month outcome of bipolar adolescents following an initial hospitalization for a manic or mixed episode. METHOD: Bipolar adolescents (N=71) were recruited during their first hospitalization for a manic or mixed episode and were evaluated using diagnostic, symptomatic, and functional assessments. Patients were also evaluated at 1, 4, 8, and 12 months after hospitalization to assess syndromic, symptomatic, and functional outcomes. Predictors of each type of outcome were identified. RESULTS: Kaplan-Meier estimates of the cumulative probabilities of syndromal, symptomatic, and functional recovery and syndromic recurrence during the first 12 months following initial hospitalization were 0.86, 0.43, 0.41, and 0.54, respectively. Only 35% of bipolar adolescents reported full medication adherence. Individual predictors of poor syndromic recovery included co-occurring attention deficit hyperactivity disorder (ADHD), anxiety disorders, and disruptive behavior disorders as well as nonadherence to psychotropic medication and lower socioeconomic levels. Co-occurring alcohol use disorders, treatment with antidepressants, and the absence of psychotherapeutic intervention predicted syndromic recurrence. Boys were more than twice as likely as girls to experience symptomatic recovery. CONCLUSIONS: Most bipolar adolescents experienced syndromic recovery following their first hospitalization. However, rates of symptomatic and functional recoveries were much lower. Future studies examining effective pharmacological and nonpharmacological treatment strategies for bipolar youth with co-occurring disorders and investigating factors that contribute to the development of substance use disorders and treatment adherence in bipolar youth are necessary to improve outcome. PMID- 17403972 TI - A randomized controlled trial of family therapy and cognitive behavior therapy guided self-care for adolescents with bulimia nervosa and related disorders. AB - OBJECTIVE: To date no trial has focused on the treatment of adolescents with bulimia nervosa. The aim of this study was to compare the efficacy and cost effectiveness of family therapy and cognitive behavior therapy (CBT) guided self care in adolescents with bulimia nervosa or eating disorder not otherwise specified. METHOD: Eighty-five adolescents with bulimia nervosa or eating disorder not otherwise specified were recruited from eating disorder services in the United Kingdom. Participants were randomly assigned to family therapy for bulimia nervosa or individual CBT guided self-care supported by a health professional. The primary outcome measures were abstinence from binge-eating and vomiting, as assessed by interview at end of treatment (6 months) and again at 12 months. Secondary outcome measures included other bulimic symptoms and cost of care. RESULTS: Of the 85 study participants, 41 were assigned to family therapy and 44 to CBT guided self-care. At 6 months, bingeing had undergone a significantly greater reduction in the guided self-care group than in the family therapy group; however, this difference disappeared at 12 months. There were no other differences between groups in behavioral or attitudinal eating disorder symptoms. The direct cost of treatment was lower for guided self-care than for family therapy. The two treatments did not differ in other cost categories. CONCLUSIONS: Compared with family therapy, CBT guided self-care has the slight advantage of offering a more rapid reduction of bingeing, lower cost, and greater acceptability for adolescents with bulimia or eating disorder not otherwise specified. PMID- 17403973 TI - Neural responses to happy facial expressions in major depression following antidepressant treatment. AB - OBJECTIVE: Processing affective facial expressions is an important component of interpersonal relationships. However, depressed patients show impairments in this system. The present study investigated the neural correlates of implicit processing of happy facial expressions in depression and identified regions affected by antidepressant therapy. METHOD: Two groups of subjects participated in a prospective study with functional magnetic resonance imaging (fMRI). The patients were 19 medication-free subjects (mean age, 43.2 years) with major depression, acute depressive episode, unipolar subtype. The comparison group contained 19 matched healthy volunteers (mean age, 42.8 years). Both groups underwent fMRI scans at baseline (week 0) and at 8 weeks. Following the baseline scan, the patients received treatment with fluoxetine, 20 mg daily. The fMRI task was implicit affect recognition with standard facial stimuli morphed to display varying intensities of happiness. The fMRI data were analyzed to estimate the average activation (overall capacity) and differential response to variable intensity (dynamic range) in brain systems involved in processing facial affect. RESULTS: An attenuated dynamic range of response in limbic-subcortical and extrastriate visual regions was evident in the depressed patients, relative to the comparison subjects. The attenuated extrastriate cortical activation at baseline was increased following antidepressant treatment, and symptomatic improvement was associated with greater overall capacity in the hippocampal and extrastriate regions. CONCLUSIONS: Impairments in the neural processing of happy facial expressions in depression were evident in the core regions of affective facial processing, which were reversed following treatment. These data complement the neural effects observed with negative affective stimuli. PMID- 17403974 TI - Neural evidence for enhanced error detection in major depressive disorder. AB - OBJECTIVE: Anomalies in error processing have been implicated in the etiology and maintenance of major depressive disorder. In particular, depressed individuals exhibit heightened sensitivity to error-related information and negative environmental cues, along with reduced responsivity to positive reinforcers. The authors examined the neural activation associated with error processing in individuals diagnosed with and without major depression and the sensitivity of these processes to modulation by monetary task contingencies. METHOD: The error related negativity and error-related positivity components of the event-related potential were used to characterize error monitoring in individuals with major depressive disorder and the degree to which these processes are sensitive to modulation by monetary reinforcement. Nondepressed comparison subjects (N=17) and depressed individuals (N=18) performed a flanker task under two external motivation conditions (i.e., monetary reward for correct responses and monetary loss for incorrect responses) and a nonmonetary condition. After each response, accuracy feedback was provided. The error-related negativity component assessed the degree of anomaly in initial error detection, and the error positivity component indexed recognition of errors. RESULTS: Across all conditions, the depressed participants exhibited greater amplitude of the error-related negativity component, relative to the comparison subjects, and equivalent error positivity amplitude. In addition, the two groups showed differential modulation by task incentives in both components. CONCLUSIONS: These data implicate exaggerated early error-detection processes in the etiology and maintenance of major depressive disorder. Such processes may then recruit excessive neural and cognitive resources that manifest as symptoms of depression. PMID- 17403975 TI - Increased waking salivary cortisol levels in young people at familial risk of depression. AB - OBJECTIVE: Cortisol hypersecretion is one of the most reliable biological abnormalities in major depression, but it is uncertain if it represents an illness marker or a trait vulnerability to mood disorder. The present study sought to answer this question by measuring waking salivary cortisol levels in young people at familial risk of depression but with no personal history of mood disorder. METHOD: The authors studied 49 young people who had not been depressed themselves but who had a parent with a history of major depression (FH+) and a comparison group of 55 participants who had no personal history of depression and no reported depression in a first-degree relative. The authors measured the amount of cortisol secreted in saliva during the first 30 minutes after awakening on a workday and on a nonworkday. RESULTS: The amount of cortisol secreted by the FH+ subjects was greater than that of the comparison subjects on both workdays (mean=698 nmol x minutes/liter, SD=243, versus mean=550, SD=225) and nonworkdays (mean=633 nmol x minutes/liter, SD=216, versus mean=492, SD=166). The increase in cortisol secretion was not accounted for by differences in parental attachment, life events, personality, or current mental state. CONCLUSIONS: Hypersecretion of cortisol can be detected in asymptomatic individuals at genetic risk of depression and may represent an illness endophenotype. Further studies will be needed to find out if increased waking salivary cortisol levels can predict individual risk of illness and whether the increased cortisol secretion has implications for general health and cognitive function. PMID- 17403976 TI - Amphetamine-induced dopamine release: markedly blunted in cocaine dependence and predictive of the choice to self-administer cocaine. AB - OBJECTIVE: Dopamine is an important mediator of the reinforcing effects of cocaine, and alterations in dopamine function might be involved in cocaine dependence. The goals of the present study were to characterize pre- and postsynaptic dopamine function in recently detoxified cocaine-dependent subjects. Specifically, dopamine response to an acute amphetamine challenge was assessed in striatal subregions in cocaine-dependent and healthy comparison participants using positron emission tomography (PET). Furthermore, the relationship between this dopamine response and the choice to self-administer cocaine in a laboratory model of relapse was investigated. METHOD: Twenty-four cocaine-dependent participants and 24 matched healthy subjects underwent [(11)C]raclopride scans under a baseline condition and following intravenous amphetamine administration (0.3 mg/kg). Cocaine-dependent participants also completed cocaine self administration sessions in which a priming dose of cocaine was followed by the choice to either self-administer subsequent cocaine doses or receive a monetary reward. RESULTS: Cocaine dependence was associated with a marked reduction in amphetamine-induced dopamine release in each of the functional subregions of the striatum (limbic striatum: -1.2% in cocaine-dependent participants versus -12.4% in healthy subjects; associative striatum: -2.6% versus -6.7%, respectively; sensorimotor striatum: -4.3% versus -14.1%). Blunted dopamine transmission in the ventral striatum and anterior caudate was predictive of the choice for cocaine over money. CONCLUSIONS: Cocaine dependence is associated with impairment of dopamine function, and this impairment appears to play a critical role in relapse. PMID- 17403977 TI - Adverse subjective experience with antipsychotics and its relationship to striatal and extrastriatal D2 receptors: a PET study in schizophrenia. AB - OBJECTIVES: Antipsychotic medications improve psychosis but often induce a state of dysphoria in patients. Blockade of the dopamine D(2) receptors, which is thought to mediate their efficacy, has also been implicated in producing this adverse subjective experience. The authors present the first double-blind controlled study to examine the relationship between striatal and extrastriatal dopamine D(2) receptor binding potential and occupancy values and adverse subjective experience. METHOD: Patients with recent-onset psychosis (N=12) were randomly assigned to low or high doses of olanzapine or risperidone. Subjective experiences, motor side effects, and striatal and extrastriatal dopamine D(2) receptors (determined with [(11)C]raclopride and [(11)C]FLB 457 PET scans, respectively) were evaluated after 2 weeks of continuous antipsychotic treatment. RESULTS: Higher dopamine D(2) receptor occupancy and binding potentials in the striatal (dorsal and ventral), temporal, and insular regions were associated with subjective experience. The finding was confirmed with two convergent methods of analysis (region-of-interest and voxel-based statistics), and the same relationship was observed using two different dopamine receptor measures (observed binding potential values and age- and sex-corrected occupancy values). CONCLUSIONS: Higher D(2) receptor occupancy is associated with negative subjective experience in patients taking risperidone or olanzapine. These negative subjective effects may be related to the high discontinuation rates seen in usual practice. Understanding the neurobiological mechanism of these negative subjective experiences and developing antipsychotics with novel (i.e., non D(2)) mechanisms may be critical in improving the treatment of psychosis. PMID- 17403978 TI - Fronto-striatal overactivation in euthymic bipolar patients during an emotional go/nogo task. AB - OBJECTIVE: Although euthymic bipolar patients show minimal manic and depressive symptoms, they continue to show impaired emotional regulation and cognitive functioning. Few studies have directly examined the interference of emotional information with cognitive processes and its underlying cerebral mechanisms in euthymic bipolar patients. The authors examined the emotional modulation of cognitive processes and its underlying neural mechanisms in euthymic bipolar patients. METHOD: Seventeen euthymic bipolar patients and 17 healthy comparison subjects underwent functional magnetic resonance imaging (MRI) while performing an emotional and nonemotional go/nogo task. Neural responses associated with the overall task performance, as well as with the impact of emotional information on the task performance, were assessed. RESULTS: Bipolar disorder patients exhibited increased activity in the temporal cortex, specifically to emotional go/nogo conditions, as well as in the orbitofrontal cortex, the insula, the caudate nuclei, and the dorsal anterior and posterior cingulate cortices when inhibiting emotional stimuli compared with neutral stimuli. Conversely, no global attentional deficits were observed on either a behavioral or neural response level, indicated by similar task performances for all task conditions and similar brain activation patterns when comparing all the go/nogo conditions with the resting state. CONCLUSIONS: The present study provides evidence of an altered emotional modulation of cognitive processing in euthymic bipolar patients, indicated by an overactivation in ventral-limbic, temporal, and dorsal brain structures during emotional go/nogo conditions in patients relative to comparison subjects. PMID- 17403979 TI - Cerebellar development and clinical outcome in attention deficit hyperactivity disorder. AB - OBJECTIVE: Anatomic magnetic resonance imaging (MRI) studies have detected smaller cerebellar volumes in children with attention deficit hyperactivity disorder (ADHD) than in comparison subjects. However, the regional specificity and longitudinal progression of these differences remain to be determined. The authors compared the volumes of each lobe of the cerebellar hemispheres and vermis in children with ADHD and comparison subjects and used a new regional cerebellar volume measurement to characterize the developmental trajectory of these differences. METHOD: In a longitudinal case-control study, 36 children with ADHD were divided into a group of 18 with better outcomes and a group of 18 with worse outcomes and were compared with 36 matched healthy comparison subjects. The volumes of six cerebellar hemispheric lobes, the central white matter, and three vermal subdivisions were determined from MR images acquired at baseline and two or more follow-up scans conducted at 2-year intervals. A measure of global clinical outcome and DSM-IV criteria were used to define clinical outcome. RESULTS: In the ADHD groups, a nonprogressive loss of volume was observed in the superior cerebellar vermis; the volume loss persisted regardless of clinical outcome. ADHD subjects with a worse clinical outcome exhibited a downward trajectory in volumes of the right and left inferior-posterior cerebellar lobes, which became progressively smaller during adolescence relative to both comparison subjects and ADHD subjects with a better outcome. CONCLUSIONS: Decreased volume of the superior cerebellar vermis appears to represent an important substrate of the fixed, nonprogressive anatomical changes that underlie ADHD. The cerebellar hemispheres constitute a more plastic, state-specific marker that may prove to be a target for clinical intervention. PMID- 17403980 TI - A quantitative trait locus analysis of social responsiveness in multiplex autism families. AB - OBJECTIVE: Autism is a complex genetic disorder with a highly heterogeneous phenotype defined by repetitive behaviors, language deficits, and problems with reciprocal social interactions. The authors present the first genome-wide scan for a social endophenotype in autism using the Social Responsiveness Scale, which provides a quantitative measure of autistic-like behavior, primarily focused on social relatedness. METHOD: A nonparametric quantitative genome scan was performed using the Social Responsiveness Scale in a cohort of about 100 families with two or more autistic probands from the Autism Genetic Resource Exchange to identify autism loci. To determine which additional loci were detected, linkage analysis with the same autism cohort using the qualitative diagnosis of autism was performed. To assess whether there were likely to be sex-specific genetic risk factors, the cohort was stratified by the sex of affected individuals. RESULTS: The quantitative Social Responsiveness Scale genome scan identified two loci on chromosomes 11 and 17, with the highest score on chromosome 11 (z=3.22). In contrast, no linkage signals reached significance in the Autism Diagnostic Interview-Revised qualitative scan. The Social Responsiveness Scale scan with only male affected identified the same signals on chromosomes 11 and 17, as well as three other regions on chromosomes 4, 8, and 10. CONCLUSIONS: This study demonstrates the utility of the Social Responsiveness Scale quantitative endophenotype to detect autism-related genetic loci using quantitative behavioral information that may more closely relate to underlying genetic factors. PMID- 17403981 TI - Risk factors for the emergence of psychotic disorders in adolescents with 22q11.2 deletion syndrome. AB - OBJECTIVE: The 22q11.2 deletion syndrome is the most common known genetic risk factor for the development of schizophrenia. The authors conducted a longitudinal evaluation of adolescents with 22q11.2 deletion syndrome to identify early risk factors for the development of psychotic disorders. METHOD: Sixty children, 31 with 22q11.2 deletion syndrome and 29 comparison subjects with idiopathic developmental disability matched for age and IQ, underwent a baseline evaluation between 1998 and 2000; of these, 51 children (28 and 23 in the two groups, respectively) underwent follow-up evaluation between 2003 and 2005. A standardized comprehensive psychiatric, psychological, and adaptive functioning evaluation was conducted in both waves. Participants with 22q11.2 deletion syndrome were also genotyped for the catechol O-methyltransferase (COMT) Met/Val polymorphism and underwent magnetic resonance imaging scans. RESULTS: The two groups had similar baseline neuropsychiatric profiles. At follow-up, 32.1% of subjects with 22q11.2 deletion syndrome had developed psychotic disorders as compared with 4.3% of comparison subjects. In the 22q11.2 deletion syndrome group, baseline subthreshold psychotic symptoms interacted both with the COMT genotype and with baseline symptoms of anxiety or depression to predict 61% of the variance in severity of psychosis at follow-up evaluation. Lower baseline verbal IQ was also associated with more severe psychotic symptoms at follow-up evaluation. CONCLUSIONS: Genetic, cognitive, and psychiatric risk factors for the evolution of psychotic disorders in 22q11.2 deletion syndrome during adolescence were identified. Early intervention in the subgroup of children with subthreshold signs of psychosis and internalizing symptoms (especially anxiety symptoms) may reduce the risk of developing psychotic disorders during adolescence. PMID- 17403982 TI - The effects of an indigenous muscarinic drug, Betel nut (Areca catechu), on the symptoms of schizophrenia: a longitudinal study in Palau, Micronesia. AB - OBJECTIVE: This study tested the findings of a prior study indicating a therapeutic relationship between consumption of betel nut and symptoms of schizophrenia. METHOD: The subjects were 65 outpatients with diagnoses of schizophrenia or schizoaffective disorder. Symptoms rated with the Positive and Negative Syndrome Scale were compared between high- and low-consumption betel chewers in a repeated-measures design. Movement disorders were assessed with the Abnormal Involuntary Movement Scale and Simpson-Angus Rating Scale. Global health and social functioning were assessed with the Medical Outcomes Study 12-item and 36-item Short-Form Health Surveys, respectively. RESULTS: Male high-consumption betel chewers had significantly milder positive symptoms than low-consumption chewers over 1 year. Betel chewing was not associated with global health, social functioning, or movement disorders. Betel chewing was associated with tobacco use but not with cannabis or alcohol. CONCLUSIONS: These findings have clinical significance in betel-chewing regions and broader implications for theory of muscarinic neurophysiology in schizophrenia. PMID- 17403983 TI - Confirmation that a specific haplotype of the dopamine transporter gene is associated with combined-type ADHD. AB - OBJECTIVE: The primary purpose of this study was to confirm the association of a specific haplotype of the dopamine transporter gene and attention deficit hyperactivity disorder (ADHD), which could be one source of the heterogeneity seen across published studies. METHOD: The authors previously reported the association of ADHD with a subgroup of chromosomes containing specific alleles of two variable-number tandem repeat polymorphisms within the 3' untranslated region and intron 8 of the dopamine transporter gene. They now report on this association in a sample of ADHD combined-type probands. RESULTS: The original observations were confirmed, with an overall odds ratio of 1.4 across samples. CONCLUSIONS: These data challenge results of meta-analyses suggesting that dopamine transporter variation does not have an effect on the risk for ADHD, and they indicate that further investigation of functional variation in the gene is required. PMID- 17403984 TI - Cost-effectiveness of schizophrenia pharmacotherapy. PMID- 17403985 TI - Reactive depression and remission rates in the STAR*D trials. PMID- 17403986 TI - The use of transference in dynamic psychotherapy. PMID- 17403987 TI - STAR*D level IV methodology. PMID- 17403988 TI - Suicide attempt following initiation of topiramate. PMID- 17403989 TI - Psychosis following initiation of zonisamide. PMID- 17403990 TI - Imagery rehearsal therapy for acute posttraumatic nightmares among combat soldiers in Iraq. PMID- 17403991 TI - Acute chorea and hyperthermia after concurrent use of modafinil and tranylcypromine. PMID- 17403992 TI - Direct effects of nicotine on contractility of the uterine artery in pregnancy. AB - Recent studies indicate that smoking/nicotine increases maternal blood pressure and decrease in uterine blood flow in pregnancy. However, the mechanisms are not fully understood. The present study was designed to test the hypothesis that nicotine exposure decreases endothelium-dependent relaxation and increases vascular contractility of the uterine artery in pregnancy. Uterine arteries were isolated from near-term ( approximately 140 days gestation) pregnant ewes. Arteries were subjected to acute (20 min) or chronic (48 h) nicotine treatment, and agonist-induced contractions and relaxations were measured in tissue bath. Endothelial eNOS was detected by immunohistochemistry in situ in arteries and by Western blotting in isolated endothelial cells. Chronic nicotine treatment produced a concentration-dependent increase in alpha(1)-adrenoceptor agonist phenylephrine-induced contractions. In contrast, the acute treatment showed no effect. Inhibition of eNOS with N(G)-nitro-L-arginine (L-NNA) significantly increased phenylephrine-induced contractions, which was abolished in uterine arteries after chronic nicotine treatment. In the presence of L-NNA, there was no significant difference in phenylephrine-induced contractions between control and nicotine-treated vessels. Chronic, but not acute, nicotine treatment significantly attenuated the calcium ionophore A23187-induced relaxations. Unlike A23187, the endothelium-independent relaxation mediated by sodium nitroprusside was not affected by nicotine. Endothelial eNOS protein levels and the phosphorylation levels of eNOS(Ser1179) were significantly decreased in nicotine treated uterine arteries. The results suggest that nicotine impairs uterine vascular function in pregnancy, which may lead to an increased vascular resistance and a decrease in uterine blood flow. PMID- 17403993 TI - In vitro pharmacology of clinically used central nervous system-active drugs as inverse H(1) receptor agonists. AB - The human histamine H(1) receptor (H(1)R) is a prototypical G protein-coupled receptor and an important, well characterized target for the development of antagonists to treat allergic conditions. Many neuropsychiatric drugs are also known to potently antagonize this receptor, underlying aspects of their side effect profiles. We have used the cell-based receptor selection and amplification technology assay to further define the clinical pharmacology of the human H(1)R by evaluating >130 therapeutic and reference drugs for functional receptor activity. Based on this screen, we have reported on the identification of 8R lisuride as a potent stereospecific partial H(1)R agonist (Mol Pharmacol 65:538 549, 2004). In contrast, herein we report on a large number of varied clinical and chemical classes of drugs that are active in the central nervous system that display potent H(1)R inverse agonist activity. Absolute and rank order of functional potency of these clinically relevant brain-penetrating drugs may possibly be used to predict aspects of their clinical profiles, including propensity for sedation. PMID- 17403994 TI - Dissemination of Acinetobacter baumannii clones with OXA-23 Carbapenemase in Colombian hospitals. AB - During 2005, 66 carbapenem-resistant isolates of Acinetobacter baumannii were collected from seven tertiary-care hospitals participating in a nationwide surveillance network in Colombia. The isolates were multidrug resistant and produced the carbapenemases OXA-23 and OXA-51. Forty-five belonged to four clones while 21 were unique pulsotypes. One clone was present in two hospitals within one city, while another had spread between two hospitals in different cities. Blood, secretions, and abdominal fluids were the most frequent sites of isolation. This is the first description of widespread dissemination of OXA-23 in South America. PMID- 17403995 TI - Efficacy of tachyplesin III, colistin, and imipenem against a multiresistant Pseudomonas aeruginosa strain. AB - An experimental study has been performed to compare the in vitro activity and the in vivo efficacy of tachyplesin III, colistin, and imipenem against a multiresistant Pseudomonas aeruginosa strain. In vitro experiments included MIC determination, time-kill, and synergy studies. For in vivo studies, a mouse model of sepsis has been used. The main outcome measures were bacterial lethality, quantitative blood cultures, and plasma levels of lipopolysaccharide, tumor necrosis factor alpha, and interleukin-6. The combination of tachyplesin III or colistin with imipenem showed in vitro synergistic interaction. A significant increase in efficacy was also observed in vivo: combination-treated groups had significantly lower levels of bacteremia than did groups treated with a single agent. Tachyplesin III combined with imipenem exhibited the highest efficacy on all main outcome measurements. These results highlight the potential usefulness of these combinations and provide therapeutic alternatives for serious infections caused by gram-negative bacteria in the coming years. PMID- 17403996 TI - Antiviral and cellular metabolism interactions between Dexelvucitabine and lamivudine. AB - Studies on cellular drug interactions with antiretroviral agents prior to clinical trials are critical to detect possible drug interactions. Herein, we demonstrated that two 2'-deoxycytidine antiretroviral agents, dexelvucitabine (known as beta-d-2',3'-didehydro-2',3'-dideoxy-5-fluorocytidine, DFC, d-d4FC, or RVT) and lamivudine (3TC), combined in primary human peripheral blood mononuclear (PBM) cells infected with human immunodeficiency virus 1 strain LAI (HIV-1(LAI)), resulted in additive-to-synergistic effects. The cellular metabolism of DFC and 3TC was studied in human T-cell lymphoma (CEM) and in primary human PBM cells to determine whether this combination caused any reduction in active nucleoside triphosphate (NTP) levels, which could decrease with their antiviral potency. Competition studies were conducted by coincubation of either radiolabeled DFC with different concentrations of 3TC or radiolabeled 3TC with different concentrations of DFC. Coincubation of radiolabeled 3TC with DFC at concentrations up to 33.3 microM did not cause any marked reduction in 3TC triphosphate (TP) or any 3TC metabolites. However, a reduction in the level of DFC metabolites was noted at high concentrations of 3TC with radiolabeled DFC. DFC-TP levels in CEM and primary human PBM cells decreased by 88% and 94%, respectively, when high concentrations of 3TC (33.3 and 100 microM) were added, which may influence the effectiveness of DFC-5'-TP on the HIV-1 polymerase. The NTP levels remained well above the median (50%) inhibitory concentration for HIV 1 reverse transcriptase. These results suggest that both beta-d- and beta-l-2' deoxycytidine analogs, DFC and 3TC, respectively, substrates of 2'-deoxycytidine kinase, could be used in a combined therapeutic modality. However, it may be necessary to decrease the dose of 3TC for this combination to prove effective. PMID- 17403998 TI - WITHDRAWN: Antimicrobial resistance, class 1 integrons and a novel variant of Genomic Island 1 in Salmonella isolates from Vietnam. AB - This manuscript has been withdrawn. PMID- 17403997 TI - Biochemical studies on the mechanism of human immunodeficiency virus type 1 reverse transcriptase resistance to 1-(beta-D-dioxolane)thymine triphosphate. AB - A large panel of drug-resistant mutants of human immunodeficiency virus type 1 reverse transcriptase (RT) was used to study the mechanisms of resistance to 1 (beta-d-dioxolane)thymine triphosphate (DOT-TP) and other nucleotide analogs. RT containing thymidine analog-associated mutations (TAM) or RT with a T69S-SG insertion in combination with TAM removed 3'-azido-3'-deoxythymidine-5' monophosphate or tenofovir more efficiently than DOT-monophosphate from chain terminated DNA primer/template through ATP-mediated pyrophosphorolysis. For non ATP-dependent discrimination toward DOT-TP, high levels of resistance were found for RT bearing the Q151M mutation with family mutations, while RT bearing only the M184V or the Y115F mutation conferred no resistance to DOT-TP. A lower degree of resistance to DOT-TP than to tenofovir diphosphate or carbovir-TP was found for RT containing the K65R mutation. In the present studies, 1-(beta-d dioxolane)guanine triphosphate, another nucleotide with a dioxolane sugar moiety, showed a resistance profile similar to that of DOT-TP. The results suggest that DOT, compared with other approved nucleoside analogs, is overall more resilient to mutations such as TAM, M184V, and K65R, which are commonly found in viruses derived from subjects failing multinucleoside therapy. PMID- 17403999 TI - Antimicrobial susceptibilities of Peptostreptococcus anaerobius and the newly described Peptostreptococcus stomatis isolated from various human sources. AB - Peptostreptococcus anaerobius sensu lato, currently including two closely related species, P. anaerobius and P. stomatis, is known to be more resistant than other gram-positive anaerobic cocci. We reidentified potential Peptostreptococcus isolates and tested their susceptibilities to eight antimicrobials. Notably, P. anaerobius had constantly higher values for the MIC at which 50% of the isolates are inhibited (MIC(50)) and the MIC(90) than P. stomatis. PMID- 17404000 TI - Controlled trial of a 5-day course of telithromycin versus doxycycline for treatment of mild to moderate scrub typhus. AB - New antibiotics are required to have the antibacterial activity against doxycycline-resistant Orientia tsutsugamushi. An in vitro sensitivity study showed that telithromycin was more effective than erythromycin for Rickettsia, Bartonella, and Coxiella burnetii. In this prospective, open-label, randomized trial, we enrolled patients with mild-to-moderate scrub typhus. We compared the efficacy and safety of a 5-day telithromycin therapy with those of a 5-day doxycycline therapy at Chosun University Hospital or one of its two community based affiliated hospitals (Jangheung Hospital and Cheomdan Hospital), which are all located in southwestern Korea, between September and December 2005. A total of 92 patients were randomly assigned to either the telithromycin group (n = 47) or the doxycycline group (n = 45). After the treatment, fever control time was 20.45 +/- 12.9 h in the telithromycin group and 22.60 +/- 21.44 h in the doxycycline group (P > 0.05). After the treatment, the cure rate was 100% in the telithromycin group and 97.8% in the doxycycline group (P > 0.05). Furthermore, there were no significant differences in time elapsed until such symptoms as headache, myalgia, and rash disappeared. No serious adverse events or death were noted following the treatment in both groups. There were no significant differences in adverse events. In conclusion, the efficacy and safety of a 5-day once-a-day regimen of 800 mg telithromycin were equivalent to those of a 5-day twice-a-day regimen of 100 mg doxycycline in patients with mild-to-moderate scrub typhus. Telithromycin could be considered a promising new antibacterial agent for patients with scrub typhus. PMID- 17404001 TI - Daptomycin nonsusceptibility in Staphylococcus aureus with reduced vancomycin susceptibility is independent of alterations in MprF. AB - A previous study documented the presence of mutations in mprF that accompanied the loss of daptomycin susceptibility among Staphylococcus aureus isolates following exposure to the drug. An association between the development of glycopeptide-intermediate S. aureus and daptomycin nonsusceptibility has also been recently described. We report that among three clinical S. aureus isolates which developed vancomycin heteroresistance, as well as daptomycin nonsusceptibility despite a lack of exposure to this drug, there were no mutations resulting in amino acid substitutions in MprF. PMID- 17404002 TI - Preservation of integron types among Enterobacteriaceae producing extended spectrum beta-lactamases in a Spanish hospital over a 15-year period (1988 to 2003). AB - The variable presence of integrons among extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae species (0 to 66%) is described. Association between bla(ESBL) and integrons occurred when these are linked to specific ESBL-type genes (In60 bearing ISCR1 and bla(CTX-M-9)) or when ESBL genes were superimposed onto selected plasmids carrying integrons. Some integrons were identical to those found during decades worldwide, illustrating the preservation of the genetic elements carrying them. PMID- 17404003 TI - Effects of piperaquine, chloroquine, and amodiaquine on drug uptake and of these in combination with dihydroartemisinin against drug-sensitive and -resistant Plasmodium falciparum strains. AB - Piperaquine is being developed as a long-acting component in artemisinin combination therapies. It was highly active in vitro and drug interaction studies showed that dihydroartemisinin combinations with piperaquine, chloroquine, and amodiaquine were indifferent tending toward antagonism. Competitive uptake of radiolabeled chloroquine and dihydroartemisinin in combination with other antimalarials was observed. PMID- 17404004 TI - Targeted drug-carrying bacteriophages as antibacterial nanomedicines. AB - While the resistance of bacteria to traditional antibiotics is a major public health concern, the use of extremely potent antibacterial agents is limited by their lack of selectivity. As in cancer therapy, antibacterial targeted therapy could provide an opportunity to reintroduce toxic substances to the antibacterial arsenal. A desirable targeted antibacterial agent should combine binding specificity, a large drug payload per binding event, and a programmed drug release mechanism. Recently, we presented a novel application of filamentous bacteriophages as targeted drug carriers that could partially inhibit the growth of Staphylococcus aureus bacteria. This partial success was due to limitations of drug-loading capacity that resulted from the hydrophobicity of the drug. Here we present a novel drug conjugation chemistry which is based on connecting hydrophobic drugs to the phage via aminoglycoside antibiotics that serve as solubility-enhancing branched linkers. This new formulation allowed a significantly larger drug-carrying capacity of the phages, resulting in a drastic improvement in their performance as targeted drug-carrying nanoparticles. As an example for a potential systemic use for potent agents that are limited for topical use, we present antibody-targeted phage nanoparticles that carry a large payload of the hemolytic antibiotic chloramphenicol connected through the aminoglycoside neomycin. We demonstrate complete growth inhibition toward the pathogens Staphylococcus aureus, Streptococcus pyogenes, and Escherichia coli with an improvement in potency by a factor of approximately 20,000 compared to the free drug. PMID- 17404005 TI - Spread in an Italian hospital of a clonal Acinetobacter baumannii strain producing the TEM-92 extended-spectrum beta-lactamase. AB - Clinical isolates of Acinetobacter baumannii (n = 470) were collected during a 7 year period and investigated for the genetic determinants of resistance to expanded-spectrum beta-lactams. Thirty-one isolates produced the TEM-92 extended spectrum beta-lactamase (ESBL) and were clonally related. This is the first report of A. baumannii producing a TEM-type ESBL. PMID- 17404006 TI - Strong and selective inhibitors of hepatitis B virus replication among novel N4 hydroxy- and 5-methyl-beta-L-deoxycytidine analogues. AB - Novel N(4)-hydroxy- and 5-methyl-modified beta-L-deoxycytidine analogues were synthesized and evaluated as anti-hepatitis B virus (HBV) agents. Their in vitro efficiencies were investigated in HepG2.2.15 cells stably transfected with HBV. beta-L-2',3'-Didehydro-2',3'-dideoxy-N(4)-hydroxycytidine (beta-L-Hyd4C) was most effective in reducing secreted HBV DNA (50% effective concentration [EC(50)], 0.03 microM), followed by beta-L-2',3'-dideoxy-3'-thia-N(4)-hydroxycytidine (EC(50), 0.51 microM), beta-L-2',3'-dideoxy-N(4)-hydroxycytidine (EC(50), 0.55 microM), and beta-L-5-methyl-2'-deoxycytidine (EC(50), 0.9 microM). The inhibition of the presumed target, the HBV DNA polymerase, by the triphosphates of some of the beta-L-cytidine derivatives was also assessed. In accordance with the cell culture data, beta-L-Hyd4C triphosphate was the most active inhibitor, with a 50% inhibitory concentration of 0.21 microM. The cytotoxicities of some of the 4-NHOH-modified beta-L-nucleosides were dramatically lower than those of the corresponding cytidine analogues with the unmodified 4-NH(2) group. The 50% cytotoxic concentrations for beta-L-Hyd4C in HepG2 and HL-60 cells were 2,500 microM and 3,500 microM, respectively. In summary, our results demonstrate that at least beta-L-Hyd4C can be recommended as a highly efficient and extremely selective inhibitor of HBV replication for further investigations. PMID- 17404007 TI - Mutant prevention concentrations of fluoroquinolones for Enterobacteriaceae expressing the plasmid-carried quinolone resistance determinant qnrA1. AB - The influence of qnrA1 on the development of quinolone resistance in Enterobacteriaceae was evaluated by using the mutant prevention concentration parameter. The expression of qnrA1 considerably increased the mutant prevention concentration compared to strains without this gene. In the presence of qnrA1, mutations in gyrA and parC genes were easily selected to produce high levels of quinolone resistance. PMID- 17404008 TI - Effect of topical microbicides on infectious human immunodeficiency virus type 1 binding to epithelial cells. AB - Topical microbicides (cellulose acetate 1,2 benzene dicarboxylate [CAP], PRO 2000, SPL7013, and UC781) are being investigated to reduce the sexual transmission of human immunodeficiency virus type 1 (HIV-1). These products were shown to prevent the transfer of infectious HIV-1 from urogenital and colorectal epithelial cell lines to peripheral blood mononuclear cells. However, it was unclear if the topical microbicides rendered the virus noninfectious and/or reduced the binding to the epithelial cells. To test this, epithelial cells were cultured with HIV-1 in the presence or absence of topical microbicides or their placebos. The cells were washed, RNA lysates were made, and real-time PCR was performed for HIV-1. PRO 2000 and SPL7013 significantly (P < 0.0001) reduced the amount of bound HIV-1 to the colorectal epithelial cell line across clades A, B, C, and CRF01-AE. While none of the products reduced the binding of HIV-1 clades A and C to the urogenital cell line, CAP, PRO 2000, and SPL7013 significantly (P 0.173, r = 0.1078, P > 0.744, r = 0.1, respectively). Significant coexpression of GLUT-1, Bcl-xL, and Bax could point to cooperation of these regulatory proteins in elimination due to irreversible injury, adaptation to hypoxia, reduction of further damage, and survival of colorectal cancer cells. PMID- 17404018 TI - Combination of doxorubicin and sulforaphane for reversing doxorubicin-resistant phenotype in mouse fibroblasts with p53Ser220 mutation. AB - Chemoresistance in cancer therapy is a multifactorial process, which includes alterations in drug accumulation, increased activity of gluthatione S transferases, loss of function, and mutations of p53, etc. One strategy for reversing chemoresistance is the use of chemopreventive agents alongside standard chemotherapeutic protocols. Sulforaphane is one of the most promising chemopreventive agents. Sulforaphane inhibits cell proliferation and induces apoptosis in different tumor cell lines. Its proapoptotic potential could make it effective either alone or in combination with other therapeutic strategies in reversing chemoresistance. We investigated the effects of sulforaphane on mouse fibroblasts bearing a different p53 status (wild-type, knockout, mutated) for understanding whether its activity is prevented by a mutated p53 status. p53 knockout fibroblasts from newborn mice transfected with the p53(Ser220) mutation, observed in different human cancers, were used as a model of mutated p53 status. Moreover, since p53(Ser220) mutation fibroblasts showed a doxorubicin-resistant phenotype, we treated the cells with a combination of doxorubicin plus sulforaphane. Taken together, our results suggest that a mutated p53 status did not prevent the induction of apoptosis by sulforaphane and that sulforaphane was able to reverse the resistance to doxorubicin. The association of sulforaphane doxorubicin may therefore allow doxorubicin to be administered at lower doses, thereby reducing its potential toxicity. PMID- 17404019 TI - Aromatase expression was not detected by immunohistochemistry in endometrial cancer. AB - Several studies suggested that aromatase could play an important role in tumor progression and prognosis in endometrial cancer because androstenedione is converted to estrogen by the enzyme. For better understanding of the aromatase expression in endometrial cancer and its relation to diverse clinicopathological parameters, we conducted this study. This study was carried out with 141 endometrial cancer patients, all of whom had undergone operations in our institution from 1993 to 2002. Paraffin-embedded tissue blocks were sectioned and immunostained with monoclonal antiaromatase antibody using human placental tissue as positive control. Clinicopathological variables of all patients were also reviewed. Despite quite a high aromatase expression in positive control, there was no endometrial cancer specimen showing the enzyme expression. Our result, although needs further investigation on the cause of the difference from other studies, suggested that aromatase might not have an important role in endometrial cancer. PMID- 17404020 TI - ER stress induces the expression of Jpk, which inhibits cell cycle progression in F9 teratocarcinoma cell. AB - Jopock (Jpk), a transacting factor associated with the position-specific regulatory element of murine Hoxa-7, has shown to induce cell death in both prokaryotic and eukaryotic cells when introduced and overexpressed. Since Jpk protein harbors a transmembrane domain (TM) and a putative endoplasmic reticulum (ER) -retention signal at the N terminus, a subcellular localization of the protein was analyzed after fusing it into the green fluorescent protein (GFP). Both N-term- (Jpk-EGFP) and C-term-fused Jpk (EGFP-Jpk) showed to be localized in the ER when analyzed under the fluorescence microscope after staining the cells with ER- and Mito-Tracker. Through deletion analysis TM turned out to be important for ER localization of Jpk. When flow cytometric analysis was performed, both cells expressing Jpk-EGFP and EGFP-Jpk led cell cycle arrest and subsequent apoptotic cell death. In order to see whether Jpk is expressed during ER stress-mediated apoptosis, F9 cells were treated with DTT, an ER stress inducer. In the presence of 4 mM of DTT, about 50% of cells died strongly expressing Jpk (sevenfold) as well as Grp78, a molecular chaperone, and CHOP-10, a well-known apoptotic protein. When cells were transfected with both pEGFP-Jpk and pJpk-EGFP, cell cycle progression was interrupted compared to those of control cells. In summary, excess ER stress upregulated the expression of Jpk, which seemed to inhibit the cell cycle progression. These results altogether suggest that Jpk could be a useful cell death-triggering molecule applicable for cancer therapy as well as a useful target molecule for the treatment of certain neurodegenerative diseases caused by ER stress. PMID- 17404021 TI - Akt involvement in paclitaxel chemoresistance of human ovarian cancer cells. AB - Paclitaxel (taxol) is extensively used for chemotherapy of various cancers including ovarian cancer. Although paclitaxel induces apoptosis in cancer cells, its exact mechanism of action still remains to be determined. Akt mediates survival signals which preserve various cancer cells from apoptosis pathway. Thus, Akt is considered an exciting target for therapeutics. Here, we demonstrated that inhibition of Akt increases the efficacy of the paclitaxel induced apoptosis in SKOV3 and PA-1 human ovarian cancer cells. The sensitivity to paclitaxel of SKOV3 and PA-1 cells was examined using the MTT assay. At a concentration of 30 nM, PA-1 cells were more sensitive to paclitaxel than SKOV3 cells. Apoptosis was accompanied by the release of cytochrome c into the cytoplasm and cleavage of poly (ADP-ribose) polymerase (PARP). To further elucidate the mechanism of apoptosis by paclitaxel, we compared the levels of phosphorylation of Akt between paclitaxel-resistant SKOV3 cells and paclitaxel sensitive PA-1 cells. The higher level of phosphorylated Akt was shown in SKOV3 cells than in PA-1 cells. Interestingly, the treatment of paclitaxel decreased the amount of phosphorylated Akt in a time-dependent manner in both cell lines. Furthermore, inhibition of Akt by specific phosphatidyinositol-3-kinase (PI3K) Akt inhibitors (Wortmannin, and LY294002) synergistically increased the efficacy of the paclitaxel-induced apoptosis in both cell lines. These results suggest that the addition of the Akt inhibitor may increase the therapeutic efficacy of paclitaxel for patients with ovarian cancer. PMID- 17404022 TI - Expression of leptin, leptin receptor, and hypoxia-inducible factor 1 alpha in human endometrial cancer. AB - Recent studies suggested that Ob (Ob) and its receptor (ObR) could be involved in the pathogenesis of various human malignancies, among others in endometrial cancer. Moreover, hypoxia, which is associated with solid tumors, might stimulate, through hypoxia-inducible factor 1alpha (HIF-1alpha), expression of Ob and ObR. In this article, we analyzed by immunohistochemistry the expression of Ob, ObR, and HIF-1alpha in 60 cases of human endometrial cancer tissues as well as in 25 cases of normal endometria. Additionally, we assessed correlations among studied proteins as well as relationships with selected clinicopathological features of endometrial cancer. Immunoreactivity for Ob, ObR, and HIF-1alpha protein was observed in 56.7%, 30.0%, and 78.3% of endometrial cancers, respectively. The expression of HIF-1alpha showed a significant positive correlation with Ob (P < 0.0001, r = 0.573) and ObR (P = 0.020, r = 0.299). Moreover, we noted positive correlation between Ob and ObR (P = 0.001, r = 0.429). No statistically significant relationship was revealed between Ob, ObR, and HIF-1alpha protein in regard to patient's age, histological grade, and extent of tumor growth (pT). In conclusion, HIF-1alpha, which is related to tissue hypoxia in endometrial cancer, seems to be associated with overexpression of Ob and ObR. Ob could exert autocrine effect to stimulate endometrial cancer progression. Thus the autocrine Ob loop should be taken into consideration as a novel potential target in endometrial cancer prevention and treatment. PMID- 17404023 TI - The cyclooxygenase-2 selective inhibitor celecoxib suppresses proliferation and invasiveness in the human oral squamous carcinoma. AB - Cyclooxygenease-2 (COX-2) expression is a critical factor in inflammation, and plays an important role in defense against exogenous stimuli, while overexpression of COX-2 causes cells to exhibit changes in tumor phenotype. This article attempted to determine the mechanisms underlying the chemopreventive effects of celecoxib on cellular level events, in order to characterize the effects of celecoxib with regard to human oral squamous cell carcinoma (OSCC) cell growth and invasion/migration. In order to determine COX-2 expression levels, we used an OSCC cell line established from surgically resected specimens of an untreated primary OSCC of the tongue, and used reverse transcription polymerase chain reaction (RT-PCR) and Western blot analyses with anti-COX-2 monoclonal antibodies. The YD-10B cells represented a highly invasive OSCC cell line, which was found to express the COX-2 protein. Celecoxib inhibited the growth of this OSCC cell line, in a time- and dose-dependent manner. This reduction in cell proliferation was associated with the upregulation of the cyclin-dependent kinase (CDK) inhibitors, p27. In addition, 10 uM celecoxib inhibited cell invasion/migration through the type I collagen matrix by approximately 40% within 24 h. The results of zymography reveal that, in the presence of 10 muL celecoxib, both MMP-2 and MMP-9 enzyme activity decreased by approximately 30-40%. The current in vitro study indicated that the inhibition of proliferation and invasion/migration in OSCC cell line by the COX-2-specific inhibitor, celecoxib, results in anticancerous effects via a variety of cellular and molecular mechanisms. This article also supports the notion that the COX-2 inhibitor may be useful in the inhibition and/or prevention of metastasis. PMID- 17404024 TI - The epidermal growth factor receptor tyrosine kinase inhibitor ZD1839 (Iressa) suppresses proliferation and invasion of human oral squamous carcinoma cells via p53 independent and MMP, uPAR dependent mechanism. AB - Oral squamous cell carcinomas (OSCCs) are characterized by a marked propensity for local invasion and dissemination to cervical lymph nodes. Overexpression of the epidermal growth factor receptor (EGFR) and high levels of certain matrix metalloproteinases (MMPs) have been implicated in the development of squamous cell carcinoma of oral cancer. ZD1839 (Iressa) is a quinazoline derivative that selectively inhibits the EGFR tyrosine kinase activity and is clinically used for cancer patients. This article attempted to determine the mechanisms underlying the effects of ZD1839 on the cellular level, and to characterize the effects of ZD1839 with regard to human OSCC cell growth and invasion/migration. The YD-10B cells represent a highly invasive human OSCC cell line, which has a frame shift p53 mutation. ZD1839 inhibited the growth of the cell line in a time- and dose dependent manner. Cell cycle kinetic analysis demonstrated that ZD1839 induces a delay in cell cycle progression and a G1 arrest. This induction of a G1 cell cycle arrest was associated with the upregulation of cyclin-dependent kinase inhibitors (CDKI) p27(KIP1) and p21(CIP1/WAF1). The upregulation of CDKI in ZD1839 treated cell lines may be mediated by a p53-independent and hnRNPC1/C2 dependent pathway. In addition, 100 nM ZD1839 demonstrated that both MMP-2 and MMP-9 enzyme activity were decreased by approximately 25-30%. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that ZD1839 downregulated the uPAR mRNA level. These results might be associated with the reduction of MMP-2 and MMP-9 activities. The current in vitro study indicates that the inhibition of proliferation and invasion/migration in OSCC cell lines by ZD1839 results in an anticancer effect via multiple cellular and molecular mechanisms, and suggests that ZD1839 may be useful in inhibiting and/or preventing metastasis. PMID- 17404025 TI - Role of vascular endothelial growth factor-D (VEGF-D) on IL-6 expression in cerulein-stimulated pancreatic acinar cells. AB - Cerulein pancreatitis is similar to human edematous pancreatitis with dysregulation of the digestive enzyme production and cytoplasmic vacuolization, the death of acinar cells, edema formation, and an infiltration of inflammatory cells into the pancreas. Our preliminary proteomic analysis showed increased expression of vascular endothelial growth factor-D (VEGF-D) in cerulein (10(-8)M) treated pancreatic acinar cells. We hypothesized that VEGF-D may have a role in cerulein-induced cytokine expression in pancreatic acinar cells. This article was designed to elucidate whether cerulein induces VEGF-D expression in pancreatic AR42J cells, and whether VEGF-D expression is related to interleukin-6 (IL-6) expression by transfecting antisense oligonucleotide (AS ODN) for VEGF-D into pancreatic acinar AR42J cells. Cerulein-induced AP-1 activation was monitored in the cells transfected with AS ODN or sense ODN (S ODN) for VEGF-D. mRNA expression of IL-6 was determined by RT-PCR analysis. AP-1 activation was assessed by electrophoretic mobility shift assay (EMSA). VEGF-D expression was assessed by Western blotting. Cerulein induced IL-6 expression and VEGF-D expression in AR42J cells. Cerulein-induced AP-1 activation and IL-6 expression was inhibited in the cells transfected with AS ODN for VEGF-D as compared to S ODN. Cerulein-induced VEGF-D expression may have a role in AP-1 activation and IL 6 expression in pancreatic acinar cells. PMID- 17404026 TI - Lack of association of the cyclooxygenase-2 and inducible nitric oxide synthase gene polymorphism with risk of cervical cancer in Korean population. AB - Currently, many studies have shown that nitric oxide (NO) and prostaglandin (PG) are main inflammatory mediators involved in a variety of pathophysiological processes including inflammation and carcinogenesis. In this article, we explored the possible association of polymorphisms, of two representative inflammatory mediators, with the risk for cervical cancer. This study included 176 cases of histologically confirmed invasive cervical cancer, and 172 healthy controls. Allele frequency of 12 single-nucleotide polymorphisms (SNPs) of cyclooxygenase-2 (COX-2) and COX5 of the inducible nitric oxide synthase (iNOS) genes in 43 different normal populations were analyzed. We found that 14 of 17 showed a monomorphic or minimal minor allele frequency; therefore, we did not continue with additional analysis. Three SNPs (2 for COX-2, 1 for iNOS) were chosen for the study. Genotyping of three SNPs (SNP-rs5275 in the untranslated region of exon 10 and rs5277 in the coding region of exon 3 of COX-2, and iNOS Ser(608)Leu allele C/T polymorphism within exon 16 of the iNOS reductase domain) was performed. No significant increase was found in any of the genotypes of the COX-2 or iNOS in the cancer group. We investigated the possible correlation between the genotypes and the clinicopathologic parameters of cervical cancer. No significant association of the genotypes studied was found with respect to clinical stage, lymph node (LN) status, histologic type, or parametrial invasion. Our data did not reveal an association between COX-2 and iNOS polymorphisms with cervical cancer in Korean women. PMID- 17404027 TI - Increased cyclooxygenase-2 expression associated with inflammatory cellular infiltration in elderly patients with vulvar cancer. AB - As the relationship between inflammation and carcinogenesis grows stronger, the role of cyclooxygenase-2 (COX-2) and epidermal growth factor (EGFR) has been highlighted in the pathogenesis and progression of human cancer. In view of the fact that vulvar cancer is characterized by precancerous inflammatory changes in elderly patients, the expressions of COX-2 and EGFR are expected to show different patterns of distribution according to age and other prognostic factors. To verify whether there was a relationship between their expression and clinicopathologic parameters in vulvar cancer, we investigated the inflammatory cellular infiltration and the expression of COX-2 and EGFR by immunohistochemical analysis. Eleven of 19 samples (57.8%) were stained positive for COX-2, and 17 (89.4%) for EGFR. The portion of inflammatory cellular infiltration in adjacent normal tissue was also higher in the older age group, and showed a strong correlation with COX-2 positivity (P = 0.002). Furthermore, COX-2 expression was significantly more frequent in patients over 60 years of age compared to those under 50 years (P = 0.009). COX-2 expression was noted to be high in moderate and well-differentiated cases, whereas, poorly differentiated carcinoma was negative for COX-2 expression (P = 0.023). However, EGFR expression was not differently distributed on the basis of stage, age, tumor grading, or presence of lymph node metastasis. Our article suggests that vulvar cancer in elderly patients may be associated with inflammation, and thus with increased COX-2 expression. In light of these findings, a clinical trial designed to assess the addition of COX-2 targeted therapy to conventional treatment in vulvar cancer would be helpful for consideration of additional treatment options and possibly avoiding the serious surgical morbidity in elderly patients. PMID- 17404028 TI - Viability of a human melanoma cell after single and combined treatment with fotemustine, dacarbazine, and proton irradiation. AB - Viability of human HTB140 melanoma cells after being exposed to fotemustine (FM) and dacarbazine (DTIC) as well as to proton irradiation was studied. Effects of 100 and 250 microM drugs were assessed after incubation of 6, 24, 48, 72, and 96 h. Irradiations were performed with 62 MeV therapeutic protons, delivering to the cell monolayer single doses of 2, 4, 8, 12, and 16 Gy. Viability was evaluated 7 days after irradiation. Inactivation level was estimated using microtetrasolium (MTT) and sulforhodamine B (SRB) assays. Combined effects of each drug and protons, were carried out using the same drug concentrations. Proton doses applied were those used in therapy, that is, 12 and 16 Gy. With the increase of drug concentration or irradiation dose, level of cell inactivation reached approximately 60%, 48 h after drug treatment or 7 days after irradiation at 16 Gy. Considering the rate of drug concentrations used, as well as the level of doses applied, it appears that HTB140 cells are more resistant to proton irradiation than to alkylating agents tested. The combined treatment with FM or DTIC and protons did not show significant changes of cell viability as compared to the effects of single agents. Since the time point for measuring cumulative effects of drug and irradiation was 48 h post irradiation, it seems that the obtained level of viability could be attributed primarily to the effects of drugs. PMID- 17404029 TI - Response of a human melanoma cell line to low and high ionizing radiation. AB - Effects of single irradiation with gamma rays and protons on human HTB140 melanoma cell growth were compared. Exponentially growing cells were irradiated close to the Bragg peak maximum of the unmodulated 62 MeV protons, as well as with (60)Co gamma rays. Applied doses ranged from 8 to 24 Gy. Viability of cells and proliferation capacity were assessed 7 days after irradiation. Induction of apoptosis and cell cycle phase redistribution were observed 6 and 48 h after irradiation. Significant inhibitory effects of both irradiation qualities were detected 7 days after irradiation. Important reduction of HTB140 cell viability was observed after irradiation with protons. Almost linear and highly significant (P < 0.001) decrease of cell proliferation was observed 7 days after irradiation with gamma rays and protons, as compared to nonirradiated controls. Protons induced apoptosis, both 6 and 48 h after irradiation. With the increase of post irradiation incubation time, number of apoptotic cells decreased. Exposure of HTB140 cells to gamma rays did not provoke apoptotic cell death. Important number of cells in G1-S phase, detected by the cell cycle phase redistribution analyses, suggested high metabolic activity of irradiated melanoma cells within the first 48 h. Both irradiation qualities caused modest G2-M arrest 6 and 48 h after irradiation, thus supporting results that illustrated high radioresistance of HTB140 cells. PMID- 17404030 TI - Effect of paclitaxel on intracellular localization of c-Myc and P-c-Myc in prostate carcinoma cell lines. AB - The proto-oncogene c-myc is involved in multiple cell pathways with opposite effects on cell outcome of death or proliferation. It has been proposed that these different roles depend on the sequestration of c-Myc protein in cellular compartments and/or its phosphorylation. We speculated that subcellular localization of c-Myc protein and of its phosphorylated form (P-c-Myc) could have a role in the different response to paclitaxel (PTX) in two prostate carcinoma cell lines, PC3 and DU145, which undergo either multinucleation or c-myc dependent apoptosis, respectively. c-myc is amplified only in PC3, but a similar extent of c-Myc phosphorylation was observed in both cell lines after PTX treatment. We found that PTX-induced upregulation of c-myc in DU145 cells, not occurring in PC3 cells, cannot be ascribed to a different protein localization, and that a comparable c-Myc and P-c-Myc nuclear translocation occurs in both cell lines after drug treatment. Thus, subcellular localization of c-Myc and P-c-Myc is not crucial in determining the mode of cell death in these prostate carcinoma cell lines. PMID- 17404031 TI - Erufosine: a membrane targeting antineoplastic agent with signal transduction modulating effects. AB - The ether lipid analog erufosine (erucylphospho-N,N,N,-trimethylpropylammonium, ErPC3) has high activity against leukemic cells without affecting the normal hematopoiesis. It belongs to the group of alkylphosphocholines (APC) that are inhibitors of protein kinase C and phospholipase C. However, the mechanism of action of erufosine remains rather unclear. We focused on combination effects with the tyrosine kinase inhibitor imatinib mesylate (gleevec, former STI-571 or CGP-57148) against two chronic myeloid leukemia (CML)-derived cell lines (K-562 and BV-173). The influence of erufosine on proteins involved in the phosphatidylinositol-3-phosphate pathway and on expression of the retinoblastoma protein Rb was studied, the latter being a key component for cell cycle entry and progression in mammalian cells. The consecutive treatment of K-562 and BV-173 cells with erufosine (2.5, 5, 15, 30 microM) and imatinib mesylate (0.05, 0.1 microM) led to synergism as measured by the MTT-dye reduction assay and this is reason to hypothesize that such combinations could be beneficial for relapsed patients with drug-resistant disease. Whole cell lysates from K-562 and BV-173 were investigated for the expression of Rb, PKB/Akt, pAkt, and p27 by Western blot. Erufosine caused decreases of pAkt and CML fusion protein p210 (BCR-ABL) protein expression, but induced the Rb protein expression in K-562 cells. A parallel increase in p27 level was observed after 24 and 48 h treatment. These alterations in signal transduction could be an explanation for the drug interaction found. Furthermore, Rb is a substrate of caspases and is cleaved during apoptosis as already evidenced for BV-173 cells. Our experimental findings suggest that erufosine acts through induction of changes in protein signaling and especially through Rb induction. This unique mode of action makes it an attractive partner for combination therapies, for example, in combination with imatinib mesylate for treatment of CML. PMID- 17404032 TI - SHP-1 tyrosine phosphatase in human erythrocytes. AB - SHP-1 is a SH2-domain containing protein Tyr-phosphatase expressed in hematopoietic cell lines, which is hypothesized to play a negative role in signal transduction. In human erythrocytes, the phospho-Tyr level of proteins, mainly transmembrane band 3, is closely controlled by the antithetic activity of Tyr protein kinases and phosphatases, resulting in a dephosphorylated state. Only after particular stimuli, as with oxidizing agents, diamide or pervanadate, or thiol alkylating compound, N-ethyl maleimide (NEM), Tyr-phosphorylation of band 3 can be triggered, inhibiting Tyr-phosphatase action and inducing erythrocyte membrane reorganization. We demonstrate that, in human erythrocytes, SHP-1 is present in membranes from resting cells, but in 5% of the protein amount. Interestingly, this amount increases up to threefold following NEM treatment of intact cells, whereas diamide and pervanadate do not alter the normal protein location. In addition, SHP-1 translocation from cytosol to membrane is not affected by band 3 P-Tyr level, because it is not mediated by the SH2-P-Tyr recruitment mechanism, and localizes into the cytoskeletal compartment. Band 3 is the target of SHP-1, which dephosphorylates Tyr 8, 21, and 904. These findings support the idea that, in human erythrocytes, the normal level of Tyr phosphorylation of membrane protein, mainly band 3, must be downregulated. We hypothesize that the presence of both SHP-2 and SHP-1 ensures band 3 dephosphorylation in different conditions: SHP-2, through interaction of its SH2 domain/s to P-Tyr protein, is regulated by the band 3 Tyr-phosphorylation level; SHP-1 may be involved by simple membrane rearrangement. PMID- 17404033 TI - Is the distribution of selenium and zinc in the sublocations of spermatozoa regulated? AB - In the sperm nuclei, of mammalian species selenium has been found only in the form of sperm nuclei glutathione peroxidase (snGPx) where it is most likely bound to the chromatin of spermatozoa. Over 80% of selenium in sperm is bound to the selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) in the midpiece of rat sperm. Zinc in sperm is mainly contained in the outer dense fiber (ODF) proteins of the flagella of mammalian spermatozoa. In the sperm nuclei, zinc is predominately located in the chromatin to the protamine proteins. In order to investigate if the insertion of zinc and selenium in sperm chromatin is regulated, the element concentrations were determined in equine spermatozoa and purified sperm nuclei. We found a significant positive correlation between the selenium concentration in equine spermatozoa and sperm nuclei. The same finding was obtained for the zinc concentration in spermatozoa and sperm nuclei. The results assume that the distribution of selenium and zinc in spermatozoa is regulated by cell signaling pathways and in this way determining the selenium and zinc amount in the chromatin of spermatozoa. PMID- 17404034 TI - Immune complexes induce monocyte survival through defined intracellular pathways. AB - Monocytes recruitment and survival at sites of inflammation are determinant for the persistence of inflammatory reactions. Immune-complexes (ICs), whose tissue deposition is involved in a variety of autoimmune diseases, activate monocytes through the interaction with Fcgamma-receptor triggering the secretion of several inflammatory modulators and favoring their tissue accumulation by inhibiting the apoptosis. To elucidate the intracellular pathways governing this process, on the basis of our previous findings regarding the dose-dependent inhibition of apoptosis in IC-activated monocytes, we have investigated the role of PI3K/Akt pathway, MAP kinases, nuclear factor-kappaB (NF-kappaB), and caspase 3, 8, and 9. Here we show that IC-activated monocytes underwent apoptosis at a rate comparable to that of resting monocytes in the presence of LY294002, a selective inhibitor of PI3K, as well in the presence of Akt inhibitor, PD98059 inhibitor of ERK1/2, and SB203580 inhibitor of p38. Moreover, IC-triggered phosphorylation of Akt, ERK1/2, and p38 MAP kinase was demonstrated on Western blot analysis. SN50, an inhibitor of NF-kappaB translocation and BMS345541, a specific inhibitor of IKK, also abolished the apoptosis protection conferred by ICs. In parallel, ICs induced an increase in NF-kappaB activation, as shown by EMSA, together with the expression of XIAP, as shown by Western blot, though indicating that in monocytes IC protection from apoptosis is NF-kappaB dependent. Finally, the activity of caspase 3, 8, and 9 resulted inhibited in IC-activated monocytes. These results disclose a signaling route triggered by ICs which can be involved in the pathophysiology of inflammatory diseases and can represent a target for therapy of IC-mediated diseases. PMID- 17404035 TI - Inhibition of serine-threonine protein phosphatases in monocyte chemoattractant protein-1 expression in Helicobacter pylori-stimulated gastric epithelial cells. AB - The reversible phosphorylation of proteins controlled by protein kinases and protein phosphatases is a major mechanism that regulates a wide variety of cellular processes, such as inflammation. It has been reported that the activity of at least 30% of all proteins can be regulated by phosphorylation in eukaryotic cells. Among these proteins, mitogen-activated protein kinases (MAPK) and several transcription factors play pivotal roles in inflammation. We previously demonstrated that Helicobacter pylori in a Korean isolate (HP99) induced proinflammatory chemokine expression by activating MAPK and transcription factors, nuclear factor-kappaB (NF-kappaB), and activator protein-1 (AP-1) in gastric epithelial AGS cells. In an attempt to determine the role of phosphorylation-dephosphorylation in HP99-induced inflammation, we analyzed the expression of protein phosphatases, the activation of MAPK and transcription factors, and the production of chemokine MCP-1 in AGS cells stimulated with HP99 (at a bacteria-cell ratio of 300:1) and cultured in the presence or absence of a nonspecific serine-threonine protein phosphatase inhibitor, okadaic acid (OA). Our results showed that HP99 induced the expression of protein phosphatases, PP1 and PP2A in AGS cells as early as 30 min. HP99 induced the activation of MAPK and AP-1, and the production of monocyte chemoattractant protein-1 (MCP-1), which were augmented by pretreatment with 100 nM of OA. Gastric epithelial cells induced the expression of PP1 and PP2A in response to HP99 presumably as a defense mechanism against inflammatory chemokine expression by inhibiting the activation of MAPK and AP-1. PMID- 17404036 TI - Cardioprotective action of urocortin in early pre- and postconditioning. AB - Pre- and postconditioning are powerful endogenous adaptive phenomenon of the organism whereby different stimuli enhance the tolerance against various types of stress. Urocortin (Ucn), member of the corticotropin-releasing factor (CRF) family has potent effects on the cardiovascular system. The aim of this article was to investigate the action of Ucn on cultured cardiomyocytes in the process of pre- and postconditioning. Isolated neonatal rat ventricular myocytes were preconditioned with adenosine, simulated ischemia, and Ucn (10-min treatment followed by 10-min reperfusion/recovery). For detecting the effect of alternative types of preconditioning, necrosis enzyme (lactate dehydrogenase [LDH]) release, vital staining (trypan blue), and ratio of apoptosis/necrosis were examined after cardiac cells were exposed to 3-h sustained ischemia and 2-h reperfusion. Same parameters were measured in the postconditioned groups (30- or 60-min ischemia followed by postconditioning with 10-min ischemic stimulus or Ucn and 2-h reperfusion). Cells exposed to 3-h ischemia followed by 2-h reperfusion were shown as control. Our results show that LDH release a number of trypan blue stained dead cells and the ratio of apoptotized and necrotized cells was decreased in all preconditioned groups compared with control group. In postconditioned groups LDH content of culture medium, trypan blue-positive cardiomyocytes, and the rate of apoptotic/necrotic cells was reduced contrasted with non-postconditioned group. We can conclude that preconditioning with Ucn induced such a powerful cell protective effect as adenosine and ischemia. Furthermore, postconditioning with Ucn after 60-min ischemia was more cardioprotective than ischemic postconditioning. PMID- 17404037 TI - Novel involvement of the immunomodulator AS101 in IL-10 signaling, via the tyrosine kinase Fer. AB - Interleukin-10 (IL-10) plays a major proliferative role in many tumors, and activates the transcription factor Stat3 by tyrosine phosphorylation. The immunomodulator ammonium trichloro (dioxoethylene-o,o') tellurate (AS101) has a direct antitumor activity, and is able to sensitize several tumors to chemotherapy, by inhibiting the tumor IL-10 autocrine loop. The tyrosine kinase Fer is essential for the proliferation of numerous malignant cell lines and in some cases was related to Stat3 activation. This article examined the role of AS101 in IL-10 signaling, and the correlation between Fer and Stat3, in human peripheral blood mononuclear cells (PBMC). We show that Fer was associated with Stat3 in PBMC and RAW 264.7, a macrophage cell line. Recombinant IL-10 (rIL-10) increased the tyrosine phosphorylation of Stat3, upregulated the levels of Fer, and increased the association of Fer with phosphorylated Stat3 (pYStat3). All the activities of IL-10 mentioned above were reversed by AS101. The effects conferred by AS101 were totally abolished by exogenous addition of rIL-10. These results indicate that AS101 downregulates the Stat3 IL-10 loop, and inhibits Fer association with pYStat3. We conclude that anti-IL-10 treatment using AS101, may be beneficial in certain malignancies and other pathologies in which IL-10 secretion is elevated and Stat3 is continuously phosphorylated. PMID- 17404038 TI - Expression and protective role of heme oxygenase-1 in delayed myocardial preconditioning. AB - In the study the authors aimed to demonstrate the expression and protective effect of heme oxygenase-1 (HO-1) in the delayed preconditioning (PC) on cultured myocardiac cells. Neonatal rat cardiac myocytes were exposed to ischemic (ischemic medium [IM] for 20 min) and pharmacological (adenosine, epinephrine, opioid) PC. Twenty-four hours later cells were subjected to a simulated ischemia (SI)--culturing for 3 h in IM, followed by 2-h reperfusion in normal medium--and then lactate dehydrogenase (LDH), live/death ratio, and apoptosis were measured. For demonstrating the protective role of HO-1, its enzymatic activity was competitively inhibited by administration of zinc protoporphyrin IX (ZnPPIX), and HO-1 synthesis was blocked with HO-1 siRNA. Cells in control group were cultured under normoxic conditions. In SI group, cells underwent only an SI without PC. HO 1 expression in all of the groups was demonstrated with immunostaining. Our results showed a significant decrease of LDH release, apoptosis, and cell death in PC groups versus SI group, which has been risen in ZnPPIX- and HO-1 siRNA treated groups. HO-1 immunostaining showed an appreciable HO-1 expression in PC groups, which was abolished with HO-1 siRNA administration, but not in ZnPPIX group. The results therefore suggest that HO-1 expression increases in both ischemic and pharmacological PC, and HO-1 has cellular protective effect against cell death and apoptosis in ischemia-reperfusion-induced oxidative injury. PMID- 17404039 TI - Expression of insulin signaling transmitters and glucose transporters at the protein level in the rat testis. AB - Insulin receptor substrate (IRS) proteins are key mediators in insulin signaling from the insulin receptor. It takes place through receptor-mediated tyrosine phosphorylation of IRS proteins. The aim of the present article is to demonstrate the distribution of IRS 1-3, glucose transporters 1-4 (GLUT 1-4), signal regulatory protein 1alpha (SIRP1alpha), PKB, and PI 3-kinase in the rat testis to see if signal transduction mediated by these proteins is active in testicular cells. Wistar rats were used as donors of testis tissue. Expression of these genes was studied at the protein level by using immunohistochemistry and Western blotting. IRS-1, IRS-2, GLUT 1, GLUT 2, GLUT 3, and SIRP1alpha were strongly expressed in the Sertoli cells (except GLUT 1), early spermatocytes, peritubular myoid cells, macrophage-like interstitial cells, and testicular endothelial cells in all the testes investigated by immunohistochemistry. IRS-2 was also expressed in the Leydig cells. Immunoblotting experiments demonstrated the presence of about 26-67 kDa reactive with anti- IRS-1, IRS-2, GLUT 1, GLUT 2, GLUT 3, PKB, and SIRP1alpha. The present results suggest that proteins like insulin and certain cytokines using IRS-1, IRS-2, GLUT 1, GLUT 2, GLUT 3, PKB, and SIRP1alpha in their signal transduction can have effects on the different types of testicular cells in the rat. PMID- 17404040 TI - Effect of endothelin on sodium/hydrogen exchanger activity of human monocytes and atherosclerosis-related functions. AB - The objective of this article is to investigate the influence of endothelin-1 (ET 1) on human monocyte Na(+)/H(+) exchanger (NHE) activity and on the atherosclerosis-related monocyte functions. ET-1 caused an increase in pHi and in (22)Na influx of monocytes. A reversal of ET-1 effect on pHi was observed in the presence of the NHE1 inhibitor, cariporide. In addition, the activation of NHE1 by ET-1 was mediated via protein kinase C (PKC), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase (PI3K), and NADPH oxidase. Also, a link between ET-1 and nitric oxide (NO) was observed. Furthermore, after ET-1 treatment, an increase of the adhesive capacity, the migration ability on laminin and CD36 expression of monocytes, was observed; using cariporide this increase was abolished. Our results showed that ET-1 induces a signaling pathway with the involvement of PKC, MAPK, PI3K, and NADPH oxidase where NHE1 plays a key role. ET 1 also plays a significant role in atherosclerosis-related functions of human monocytes, via NHE1 activation. PMID- 17404041 TI - Small GTPase Ras and Rho expression in rat osteoblasts during spaceflight. AB - Rat osteoblasts were cultured for 4 and 5 days aboard a space shuttle and solubilized after a 24-h treatment with 1alpha,25 dihydroxyvitamin D(3). The quantitative RT-PCR determined the mRNA levels of signaling molecules upstream and downstream Ras. The small GTPase is activated by guanine nucleotide exchange protein (GEF) and deactivated by GTPase-activating protein (GAP). When external stimuli are transduced into intracellular signals, various pathways are recruited: focal adhesion kinase (FAK) is associated with integrin-beta, and directs tyrosine phosphorylation of downstream substrates, including phospholipase C-gamma (PLC-gamma) and son of sevenless (SOS, a Ras GEF). The mRNA levels of FAK and PLC-gamma1 and -gamma2 in the flight cultures were increased 150% and 250% of the ground controls. The SOS mRNA levels in the flight cultures were increased 520% and 320% of the ground controls. Signals via G protein coupled receptors are transmitted through PLC-beta and Ras GRF (another Ras GEF). Activated Ras then stimulates Raf, mitogen-activated protein kinase (MAPK) cascades. The mRNA levels of Raf, extracellular signal-regulated protein kinase of MAPK family (ERK-1 and -2), and PLC-beta were increased during spaceflight. Rho GAP expression in the flight cultures was increased twofold of the ground controls. Since Rho GAP deactivates Rho, microgravity may suppress Rho signals, regulating actin filament rearrangement. Microgravity signals may involve two pathways (G protein-coupled receptor-mediated pathway and tyrosine phosphorylation-mediated pathway) that activate Ras, Raf, and MAPK cascades in rat osteoblasts. PMID- 17404042 TI - Protein expression in the tissues of the cardiovascular system of the rat under selenium deficiency and adequate conditions. AB - We investigate the effects of selenium in tissues of the cardiovascular system. In order to examine the expression of the heart and aorta proteins (part of them) in the homogenate of selenium-deficient (Se(-)) and selenium control rats (Se(+)), two-dimensional electrophoresis was used by means of giga gels (30 x 35 cm). After electrophoresis, the protein expression pattern of the (Se(-)) gel and (Se(+)) gel was compared. The evaluation of the protein difference was implemented by means of a computer program suitable for the analysis of protein separated by the two-dimensional electrophoresis. In this way more than 2000 proteins a gel (heart) were detected and more than 1900 protein spots were detected in the aorta fraction. Ten significant differences were found between the gel of (Se(+)) and (Se(-)) heart homogenate of the rat and more than 15 significant differences between the gel of (Se(+)) and (Se(-)) of the aorta. By means of MALDI-MS-ESI-MS some of these proteins with different expression levels were not determined until now. Of those, three proteins were detected as the alpha myosin heavy chain (alpha-MHC), myosin light chain 1 and 2 (MLC 1 and 2), and the mitochondrial enzyme creatinine kinase. First results suggest that selenium deficiency affects myocardial energy metabolism and contractile proteins. PMID- 17404043 TI - Study on the correlations among disease activity index and salivary transforming growth factor-beta 1 and nitric oxide in ulcerative colitis patients. AB - Growth factors and nitric oxide (NO) play a major role in dysregulated immune response in ulcerative colitis (UC). Recent evidence has shown increased levels of transforming growth factor-beta(1) (TGF-beta(1)) in UC and suggested an anti inflammatory effect for this factor. Based on our recent study, dysfunctional immunoregulation is present in saliva of UC patients, we hypothesized that salivary level of NO and TGF-beta(1) may differ by severity of UC and be useful to determine the activity of the disease. Thirty-seven UC patients and 15 healthy controls were enrolled and saliva samples were obtained. Truelove-Witts severity index and modified Truelove-Witts severity index were used to determine the severity of the disease. NO and TGF-beta(1) levels were detected in saliva of all patients and control subjects using enzyme-linked immunosorbent assay. A total of 21 patients had mild disease while 8 had moderate and 8 had severe colitis. Adjusted for baseline characteristics, the levels of NO and TGF-beta(1) in different groups were compared. Salivary NO and TGF-beta(1) levels were higher in UC patients comparing to controls (P < 0.00005 and P = 0.005, respectively). The levels of NO and TGF-beta(1) showed no significant differences among the severity groups (P = 0.46 and P = 0.23, respectively). NO levels linearly increased by age (Coeff = 1.5, r = 0.38, P = 0.02). Gender, extension of disease, and medical treatment did not affect NO and TGF-beta(1) levels. Although UC patients have abnormal amounts of NO and TGF-beta(1) in their saliva, their disease activity cannot be predicted by these factors, which may indicate a pathophysiologic role rather than being nonspecific inflammatory markers for TGF-beta(1) and NO. PMID- 17404044 TI - Generation of DeltaTAp73 proteins by translation from a putative internal ribosome entry site. AB - p73 belongs to a family of transcription factors, including p53 and p63, that mediate response to DNA damage and cellular stress by inducing DNA repair, cell cycle arrest, and apoptosis. TP73 gene contains two promotors and several splice variants resulting in up to 24 possible permutations of p73 proteins which underlies the complexity of the family and its regulatory mechanisms. p73 variants lacking the N-terminal, denoted as DeltaTAp73, are not transcriptionally competent and they act in a dominant negative fashion over TAp73. DeltaTAp73 isoforms can be generated by alternative promotor usage, giving rise to DeltaNp73, or alternative splicing of exons 2, 3 or 2, and 3 together. Such transcript isoforms potentially produce oncogenic proteins and they were shown to be present in primary tumors and tumor-derived cell lines. We investigated the possibility of additional mechanisms by which p73 protein could be regulated and discovered a putative internal ribosome entry site (IRES) in exon 2. Translation initiation of TAp73 mRNA results in a DeltaNp73-like peptide, thus demonstrating an additional mechanism whereby a DeltaTA p73 protein is produced from a transcript originally generated from the P1 promotor of the p73 gene. PMID- 17404045 TI - IRF-7: new role in the regulation of genes involved in adaptive immunity. AB - The interferon regulatory factor 7 (IRF-7), a member of the IRF family of transcription factors, is a key player in the innate immune response against viral infections. Constitutive expression of IRF-7 is limited to peripheral blood lymphocytes and dendritic cells while in most cell types its expression can be induced by type I interferon (INF). IRF-7 is sequestered in the cytoplasm of uninfected cells and following viral infection, double-stranded RNA (dsRNA), or toll-like receptor (TLR) signaling, it becomes phosphorylated by TBK and IKK-i kinases. Phosphorylated IRF-7 migrates in the nucleus where it can activate IFN type I genes and other interferon-stimulated genes (ISGs). Here we report that the overexpression of a constitutively active form of IRF-7 binds and positively regulates the transcriptional activity of the promotor of IRF-1 and low molecular mass polypeptide-2 (LMP-2), two proteins that play a key role in adaptive immunity. The so far unrecognized role of IRF-7 in LMP-2 stimulation points to IRF-7 as a transcriptional regulator that bridges innate and adaptive immunity. PMID- 17404046 TI - A new transcript splice variant of the human glucocorticoid receptor: identification and tissue distribution of hGR Delta 313-338, an alternative exon 2 transactivation domain isoform. AB - All human glucocorticoid receptor (hGR) isoforms are encoded by the NR3C1 gene consisting of seven core exons (exons 2-8) common to all protein isoforms. The gene has two major exon 8-9 splice variants and a 5'-UTR consisting of 11 alternative splice variants. The N-terminal region of the hGR includes a tau 1 transactivation domain that interacts with proteins in the basal transcriptional apparatus, including the TATA box-binding protein. Here, we report the existence and the tissue distribution of a novel splice variant, hGRDelta313-338, with a 26 residue (78 bp) deletion in this N-terminal region encoded by exon 2, between amino acids 313 and 338. The hGRDelta313-338 observed at the mRNA level represents a transcript variant encoding a smaller protein isoform detected by WB with a predicted deletion between the tau 1 domain and the DNA-binding domain (DBD) encoded by exons 3 and 4. Previous studies in transgenic mice showed that the removal of the entire exon 2 covering both the tau 1 transactivation domain and our deleted region produced a functional receptor albeit with an altered glucocorticoid-induced gene transcription pattern. Interestingly, the deleted residues show a number of potential phosphorylation sites including serine 317, known to be phosphorylated. It is thought that phosphorylation plays an important role in transactivation action of hGR. Thus, we hypothesize that hGRDelta313-338 represents a hGR isoform with an altered glucocorticoid-induced transactivation profile. PMID- 17404047 TI - Exploitation of host signaling pathways by B cell superantigens--potential strategies for developing targeted therapies in systemic autoimmunity. AB - Some infectious agents produce molecules capable of interacting specifically with the immunoglobulin heavy- or light-chain variable regions, independently of the conventional-binding site. They are referred to as B cell superantigens (SAgs) and include protein A of Staphylococcus aureus (S. aureus), gp120 of HIV-1, and protein L of Peptostreptococcus magnus (P. magnus). In contrast to conventional antigens, B cell superantigens interact with conserved framework regions of immunoglobulins and can target a large proportion of B cells. In experimental models, they have been demonstrated to deplete B cell subsets responsible for innate functions, namely B-1a and marginal zone (MZ) B cells. As a result, the interactions of these superantigens with host cells impair the humoral immune response. In addition to providing clues toward understanding host-pathogen interactions and microbial pathogenesis, B cell superantigens represent potential therapeutic agents that could be used to specifically modulate expansion of B cell subsets in diseased subjects. In systemic autoimmune diseases, for example, there is activation and expansion of B cells that secrete pathogenic autoantibodies. Their depletion results in clinical improvement in both experimental animals and patients. Currently, attempts are being made to specifically deplete pathogenic autoantibody-producing B cells. Since B-1a and MZ B cells have been found to be expanded in autoimmune disorders, B cell superantigens, used alone or in combination with other biological agents, may have beneficial effects in autoimmune disease management. PMID- 17404048 TI - Antineoplastic and anticlastogenic properties of curcumin. AB - Curcumin is the pigment of turmeric and has been reported as a signal transduction modulator and inhibitor of transcription factors, for example, NF kappaB. In our article we found a concentration-dependent cytotoxic activity of curcumin in a panel of eight leukemic cell lines (SKW-3, CEM, U-937, HL-60, HL 60/Dox, K-562, LAMA-84, and AR-230). Additive to synergistic interactions was recorded for combinations with bendamustine and idarubicine in SKW-3 and LAMA-84 cells. Noteworthy, in multiple myeloma cells (RPMI-8226 and U-266) a potentiation of the efficacy of bendamustine by curcumin application was found. Moreover, curcumin increased the bendamustine cytotoxicity in cultures of cells isolated from the bone marrow of a patient with non-Hodgkin's lymphoma (NHL). The increased bendamustine efficacy could be explained by NF-kappaB inhibition, because this factor is activated in many cancers, especially leukemia and multiple myeloma. Curcumin is characterized by low toxicity and was described to have a chemoprotective activity. Therefore, the level of reduced glutathione (GSH) was measured and a concentration-dependent increase of GSH levels was recorded in AR-230 and SKW-3 cells (concentration range 5-25 muM). Experiments with mice showed significant protection against cisplatin-induced chromosomal aberrations (clastogenic effect) and inhibition of mitoses in bone marrow cells. Curcumin alone caused reduction of the mitotic index. In combination with cisplatin, however, this parameter was increased when compared to cisplatin alone. Our data indicate that curcumin has pleiotropic effects on signal transduction by inhibiting transcription thus exerting antitumor activity. In addition, curcumin has protective and anticlastogenic activity by enhancing the scavenging of free radicals. PMID- 17404049 TI - Sanguinarine inhibits VEGF-induced Akt phosphorylation. AB - Angiogenesis is the process of vascular growth by sprouting of preexisting vessels. This process impacts significantly on many important disease states including cancer, diabetic retinopathy, and arthritis. Endothelial cells receive multiple information from their environment, which leads them to progress along all stages of new vessel formation. Vascular endothelial growth factor (VEGF), in particular appears to be a master regulator of this process. This molecule interacts with cellular receptors and communicates with cell nucleus through a network of intracellular signaling, most of all by activating Akt pathway. This activation accounts for many of VEGF effects, including cell survival, migration, tube formation, and promotion of NO release. Sanguinarine (SA), an alkaloid isolated from Sanguinaria canadensis, is known for its antiangiogenetic effects by suppressing basal and VEGF-induced new vessel growth. This article was aimed to evaluate the possible effect of SA (300 nM) on Akt phosphorylation in a porcine aortic endothelial cell line. The alkaloid significantly (P < 0.001) inhibited the VEGF-induced Akt increase, thus suggesting that this mode of action could be responsible, at least partially, for the antiangiogenetic effect of SA. PMID- 17404050 TI - Effect of curcumin treatment on protein phosphorylation in K562 cells. AB - Deregulation of signaling pathways is a common feature observed in human cancers and other diseases. Therefore, there is a strong need for compounds that are able to modulate or inactivate upregulated signaling events. Natural compounds extracted from plants have long been used and still present a dynamic domain in the research of new therapeutic tools. Among those molecules, curcumin was already described for its antioxidative, anti-inflammatory, and antiseptic properties. Many actions of curcumin target proteins and kinases implicated in the signaling pathways. However, the effects described depend on the treatment conditions used, as well as the cell line studied, and these features vary strongly from one study to the other. During this work, we evaluated the effect of one curcumin treatment (20 muM, 48 h) on the phosphorylation of a number of proteins and kinases in the human chronic myelogenous leukemia cell line K562. These results allow to compare the results obtained in one condition on various proteins. PMID- 17404051 TI - Dosage effects of ginkgolide B on ethanol-induced cell death in human hepatoma G2 cells. AB - Ginkgolide B is a major active component of Ginkgo biloba extracts, which has been shown to confer anticancer effects by inducing apoptosis or inhibiting oxidative stress generation. Ethanol induces a wide range of cellular toxicities, many of which have been linked to free radical generation. To further elucidate the cellular effects of ginkgolide B, we examined the dose-response effect of ginkgolide B on ethanol-induced toxicity in human Hep G2 cells. TUNEL and MTT assays revealed that ethanol (50-400 mM) induced apoptotic cell death in human Hep G2 cells, and that this effect was inhibited by low (5-25 microM) doses of ginkgolide B, but enhanced by high (50-100 microM) doses of ginkgolide B. Additional experiments revealed that ethanol treatment directly increased intracellular oxidative stress; this effect was enhanced by high doses of ginkgolide B but decreased following treatment with low concentrations of ginkgolide B. The dose-response effects of ginkgolide B on reactive oxygen species (ROS) generation were directly correlated with cell apoptotic biochemical changes including c-Jun N-terminal kinase (JNK) activation, caspase-3 activation, and DNA fragmentation. These results indicate that treatment dosage may determine the effect of ginkgolide B on ethanol-induced ROS generation and cell apoptosis, and support the notion that an appropriate dosage of ginkgolide B may aid in decreasing the toxic effects of ethanol. PMID- 17404052 TI - Attenuation of Abeta-induced apoptosis of plant extract (Saengshik) mediated by the inhibition of mitochondrial dysfunction and antioxidative effect. AB - Recently, considerable attention has been focused on dietary manipulation of oxidative and/or nitrosative damage on neuronal cells. In this article, a neuroprotective effect of plant (Saengshik) extracts was investigated. Rat pheochromocytoma (PC12), cells treated with beta-amyloid underwent apoptotic death as determined by positive in situ terminal end-labeling (TUNEL staining), decreased mitochondrial transmembrane potential, and elevated caspase-3 activity co-occurring with enhanced MDA accumulation and the reduction of GSH levels. Saengshik pretreatment attenuated beta-amyloid-induced apoptosis in PC12 cells possibly by inhibiting mitochondrial dysfunction and exerting antioxidant properties. Saengshik pretreatment inhibited the loss of mitochondrial membrane potentials and reduced the activation of caspase-3. The in vitro antioxidant activities of Saengshik extracts were verified by the 1,1-diphenyl-2 picrylhydrazyl (DPPH) method and superoxide dismutase (SOD) mimetic activity. In beta-amyloid-challenged PC12 cells, Saengshik prevented the production of ROS, decreased the level of MDA, and elevated GSH. The potential of Saengshik as one of the neuroprotective regimens has been suggested through this article, and the combination with defined pharmaceuticals or other dietary antioxidants may provide a better therapeutic or preventive advantage for the management of Alzheimer's disease. PMID- 17404053 TI - Wogonin prevents immunosuppressive action but not anti-inflammatory effect induced by glucocorticoid. AB - Glucocorticoid, such as dexamethasone, has anti-inflammatory and immunosuppressive action as major pharmacological effects. The latter action caused by lymphocyte apoptosis is not only a therapeutic effect but also an adverse reaction. Wogonin, a plant flavone found in Scutellaria baicalensis Georgi, inhibited dexamethasone-induced apoptotic changes, such as DNA fragmentation, nuclear condensation, phosphatidylserine translocation, and caspase activation in rat thymocytes. Since wogonin inhibited dexamethasone induced DNA fragmentation in a noncompetitive manner, a target of this flavone is unlikely to be an antagonist of glucocorticoid receptor. Wogonin did not only act as an inhibitor of caspases, but also protected apoptosis induced by other glucocorticoids. Since wogonin reduced one of the major pharmacological effects of dexamethasone, we examined whether this flavone diminishes the anti inflammatory action, another pharmacological effect. The anti-inflammatory action of dexamethasone was evaluated by carrageenan-induced paw edema model. Although dexamethasone significantly suppressed paw edema induced by carrageenan, wogonin had no effect on the anti-inflammatory action of dexamethasone. These results suggest that wogonin may be a useful compound to reduce the immunosuppressive side effect of glucocorticoid. PMID- 17404054 TI - The analgesic effect of Tribulus terrestris extract and comparison of gastric ulcerogenicity of the extract with indomethacine in animal experiments. AB - Tribulus terrestris has been used in traditional medicine for relieving rheumatic pain and as an analgesic plant for a long time. In this investigation the analgesic effect of methanolic extract of this plant on male albino mice was evaluated by formalin and tail flick test. Extraction of the fruits of the plant was done by two different methods (suxheletion and percolation) with methanol 80%. The percolated extract was injected intraperitoneally in mice at 50, 100, 200, 400, and 800 mg/kg. The results showed that a dose of 100 mg/kg of percolated extract had the highest significant analgesic effect compared to the control group (P < 0.01) in formalin and tail flick test. There is no significant difference in the analgesic effect of suxheleted and percolated extract. The analgesic effect of the extract was lower than morphine, 2.5 mg/kg in both tests, and higher than ASA 300 mg/kg in chronic phase of pain in formalin test (P < 0.05). Pretreatment of animal with naloxone did not change the analgesia induced by the plant extract in both tests, therefore the involvement of opioid receptor in the analgesic effect of this plant was excluded. The results of ulcerogenic studies indicate that the gastric ulcerogenecity of plant extract is lower than the indomethacin in the rat's stomach. It can therefore be concluded that T. terrestris extract has a suitable analgesic effect and further studies are required to produce a more effective product of this plant to substitute for conventional analgesic drugs. PMID- 17404055 TI - Epigallocatechin gallate dose-dependently induces apoptosis or necrosis in human MCF-7 cells. AB - The catechins, a family of polyphenols found in tea, can evoke various responses, including cell death. However, the precise molecular mechanisms of these effects are unknown. Here, we demonstrate that treatment of human MCF-7 cells with 50 microM (-)-Epigallocatechin-3-gallate (EGCG), a catechin that is highly abundant in green tea, can induce apoptotic changes, including mitochondrial membrane potential changes and activation of c-Jun N-terminal kinase (JNK), caspase-9, and caspase-3. In contrast, higher concentrations of EGCG (100-400 microM) do not induce apoptosis, but rather trigger necrotic cell death in MCF-7 cells. Investigations of the possible mechanisms underlying these differences revealed that treatment with lower concentrations of EGCG (10-50 microM) directly increased intracellular oxidative stress, while higher concentrations (100-400 microM) did not. Immunoblotting revealed that treatment of MCF-7 cells with 10-50 microM EGCG caused increases in Bax protein levels and decreases in Bcl-2 protein levels, shifting the Bax-Bcl-2 ratio to favor apoptosis, while treatment with 100 400 microM EGCG had no such effect. Moreover, we observed a dose-dependent decrease in intracellular ATP levels in cells treated with high-dose EGCG. Blockade of reactive oxygen species (ROS) generation and ATP synthesis using antioxidants and ATP synthesis inhibitors revealed that ROS and ATP play important roles to switch cell death types with apoptosis or necrosis. Collectively, these results indicate for the first time that EGCG treatment has a dose-dependent effect on ROS generation and intracellular ATP levels in MCF-7 cells, leading to either apoptosis or necrosis, and that the apoptotic cascade involves JNK activation, Bax expression, mitochondrial membrane potential changes, and activation of caspase-9 and caspase-3. PMID- 17404056 TI - Resveratrol induces apoptosis in chemoresistant cancer cells via modulation of AMPK signaling pathway. AB - Resveratrol has been reported to possess therapeutic effects for various cancers including colon cancers. In this article, the molecular basis of resveratrol with emphasis on its ability to control intracellular signaling cascades of adenosine monophosphate (AMP)-activated protein kinase (AMPK) responsible for inducing apoptosis in drug-resistant cancer cells was investigated. Recently, the evolutionarily conserved serine/threonine kinase, AMPK, emerges as a possible target molecule of cancer control. We have investigated the effects of resveratrol on apoptosis in relation to AMPK in HT-29 cells shown chemoresistant to a cancer chemotherapeutic drug, etoposide. Resveratrol exhibited a variety of molecular events in etoposide-based combination therapy in HT-29 colon cancer cells including the AMPK activation, inhibition of cell growth, induction of apoptosis, and reactive oxygen species (ROS) generation. The involvement of AMPK signaling cascade in resveratrol-based cancer therapy was clearly shown by comparing the conditions of AMPK activated states and inactivated states. We have identified ROS as an upstream regulator of AMPK. Further investigation warrants to elucidate the mechanism by which resveratrol generates ROS and AMPK activation. PMID- 17404057 TI - Antioxidative effects of plant polyphenols: from protection of G protein signaling to prevention of age-related pathologies. AB - The antioxidant potency of three natural polyphenols, resveratrol, curcumin, and genistein, was compared by using the two human models: oxymodified with H(2)O(2) and homocysteine (Hcy) G proteins in the postmortem frontal cortex (FC) membranes of age-matched control and Alzheimer's disease (AD) subjects; and Cu(2+)-induced oxidation of plasma low-density lipoproteins (LDL). In Co, 3-10 microM polyphenols dose-dependently depressed the G protein 25% stimulation induced by 10 microM H(2)O(2) or 500 microM Hcy. Resveratrol revealed significantly higher antioxidativity than curcumin or genistein. In AD, the antioxidativity of polyphenols showed no significant differences. Polyphenols (1 microM) significantly increased the LDL oxidation lag time (oxyresistance) as compared with control, the effect of resveratrol being most potent. Due to the dual antioxidant mechanism, the investigated polyphenols, particularly resveratrol, should have preferences for the preventive-therapeutic use in age-related oxidative stress-based pathologies. PMID- 17404058 TI - Jaceosidin, a pharmacologically active flavone derived from Artemisia argyi, inhibits phorbol-ester-induced upregulation of COX-2 and MMP-9 by blocking phosphorylation of ERK-1 and -2 in cultured human mammary epithelial cells. AB - Elevated levels of cyclooxygenase-2 (COX-2) and matrix metalloproteinases (MMPs) are frequently found in various types of cancerous and transformed cells, with recent studies implicating the upregulation of COX-2 and MMPs in the development of breast cancer. This article investigated the effects of jaceosidin (4',5,7 trihydroxy-3',6-dimethoxyflavone) isolated from Artemisia argyi on the upregulation of COX-2 and MMP-9 induced by the tumor promotor 12-O tetradecanoylphorbol-13-acetate (TPA) in MCF10A human breast epithelial cells (MCF10A cells). Treatment of MCF10A cells with TPA induced the upregulation of COX-2 and MMP-9, and this was attenuated by jaceosidin treatment. Jaceosidin also inhibited the invasive and migrative phenotypes of MCF10A cells induced by TPA. Furthermore, jaceosidin blocked the TPA-induced phosphorylation of extracellular signal-regulated protein kinase-1 and -2 (ERK-1 and -2), which is one of the signaling molecules regulating COX-2 and MMP. These results suggest that jaceosidin inhibits the TPA-induced upregulation of COX-2 and MMP-9 by blocking ERK-1 and -2 phosphorylation in human breast epithelial cells, which may be indicative of its chemopreventive potential. PMID- 17404059 TI - Effects of selenium diet on expression of selenoproteins in the lung of the rat. AB - In the present article the radiotracer techniques have been combined with biochemical separation procedures to investigate the effects of changes in the selenium status on the expression of the selenium-containing proteins in the lung and their subcellular fractions. Subcellular separation of the lung has been achieved by differential ultracentrifugation. The selenium-containing proteins in these compartments have been investigated by labeling of rats in vivo with (75)Se, gel electrophoretic separation of the proteins, and autoradiographic detection of the tracer. In the lung of the selenium-deficient animals, the selenium administered was used predominantly to restore the levels of the selenoproteins, while in the lung of the selenium-sufficient animals most of the selenium retained was incorporated into the glutathione peroxidase. Also, higher activity of this enzyme has been found in the lung of the selenium-sufficient animals. The differences in the specific incorporation of the element in the selenium deficiency into different compounds suggested that there are different metabolic pathways for selenium, strongly dependent on its status. PMID- 17404060 TI - Protective effects of piceatannol against beta-amyloid-induced neuronal cell death. AB - Beta-amyloid (Abeta) is a main component of senile plaques in Alzheimer's disease (AD) that induces neuronal cell death. Since reactive oxygen species (ROS) have been implicated in Abeta-induced neurotoxicity, considerable attention has recently been focused on identifying naturally occurring antioxidative phenolic phytochemicals that are able to decrease ROS levels. Piceatannol (trans-3,4,3',5' tetrahydroxystilbene), which has a structure homologous to resveratrol, is an anti-inflammatory and antiproliferative stilbene compound derived from plants. This article investigated the possible protective effects of piceatannol on Abeta induced PC12 neuronal cell death, and found that piceatannol exerted much stronger protective effects than did resveratrol. Piceatannol treatment attenuated the intracellular accumulation of ROS induced by treatment of PC12 cells with Abeta, inhibited Abeta-induced apoptotic features including internucleosomal DNA fragmentation, nucleus condensation, cleavage of poly(ADP ribose) polymerase (PARP), and activation of caspase-3. These results suggest that piceatannol blocks Abeta-induced accumulation of ROS, thereby protecting PC12 cells from oxidative stress. PMID- 17404061 TI - Jaceosidin induces apoptosis in ras-transformed human breast epithelial cells through generation of reactive oxygen species. AB - Extracts of Artemisia plants possess anti-inflammatory and antioxidative activities. Eupatilin (5,7-dihydroxy-3',4',6-tri-methoxy-flavone), a pharmacologically active flavone derived from Artemisia asiatica, was shown to inhibit phorbol ester-induced cyclooxygenase-2 expression and NF-kappaB activation in mouse skin, and also to induce cell cycle arrest in ras-transformed human mammary epithelial (MCF10A-ras) cells. In this article, we examined the ability of jaceosidin (4',5,7-trihydroxy-3',6-dimethoxyflavone) isolated from Artemisia argyi to inhibit the proliferation of MCF10A-ras cells. Jaceosidin reduced the viability of MCF10A-ras cells to a greater extent than eupatilin. Jaceosidin treatment resulted in increased intracellular accumulation of reactive oxygen species (ROS) in MCF10A-ras cells, which was blocked by the antioxidant N acetylcysteine (NAC). NAC attenuated jaceosidin-induced cytotoxicity. To better assess the proapoptotic effects of jaceosidin, we analyzed the treated cells by the flow cytometry. MCF10A-ras cells treated with jaceosidin (100 microM) exhibited the increased proportion of hypodiploid or apoptotic cells (48.72% as composed to 7.78% in control cells). Jaceosidin treatment also increased the ratio of proapoptotic Bax to the antiapoptotic Bcl-2 and induced the cleavage of caspase-3 and poly(ADP-ribose)polymerase (PARP). Moreover, jaceosidin elevated the expression of p53 and p21, while the compound inhibited the activation of ERK1/2 that is an important component of cell survival signaling. PMID- 17404062 TI - Involvement of AMPK signaling cascade in capsaicin-induced apoptosis of HT-29 colon cancer cells. AB - Adenosine monophosphate (AMP)-activated protein kinase (AMPK) is activated during ATP-depleting metabolic states, such as hypoxia, heat shock, oxidative stress, and exercise. As a highly conserved heterotrimeric kinase that functions as a major metabolic switch to maintain energy homeostasis, AMPK has been shown to exert as an intrinsic regulator of mammalian cell cycle. Moreover, AMPK cascade has emerged as an important pathway implicated in cancer control. In this article, we have investigated the effects of capsaicin on apoptosis in relation to AMPK activation in colon cancer cell. Capsaicin-induced apoptosis was revealed by the presence of nucleobodies in the capsaicin-treated HT-29 colon cancer cells. Concomitantly, the activation of AMPK and the increased expression of the inactive form of acetyl-CoA carboxylase (ACC) were detected in capsaicin-treated colon cancer cells. We showed that both capsaicin and 5'-aminoimidazole-4 carboxamide-1-beta-D-ribonucleoside (AICAR), an AMPK activator possess the AMPK activating capacity as well as apoptosis-inducing properties. Evidence of the association between AMPK activation and the increased apoptosis in HT-29 colon cancer cells by capsaicin treatment, and further findings of the correlation of the activated AMPK and the elevated apoptosis by cotreatment of AICAR and capsaicin support AMPK as an important component of apoptosis, as well as a possible target of cancer control. PMID- 17404063 TI - Epigallocatechin gallate inhibits phorbol ester-induced activation of NF-kappa B and CREB in mouse skin: role of p38 MAPK. AB - The modulation of intracellular signaling network involved in an inappropriate expression of cyclooxygenase-2 (COX-2) is a pragmatic approach for chemoprevention with a wide variety of dietary phytochemicals. Epigallocatechin gallate (EGCG), a major green tea polyphenol, is one of the most extensively investigated chemopreventive agents. Our previous study revealed that EGCG inhibited expression of COX-2 and activation of mitogen-activated protein kinases (MAPKs) in mouse skin stimulated with a prototype tumor promotor 12-O tetradecanoylphorbol-13-acetate (TPA). This study was aimed at identifying transcription factors as molecular targets of EGCG in downregulating COX-2 expression. We found that EGCG inhibited TPA-induced DNA binding of NF-kappaB and CREB in mouse skin in vivo. EGCG also suppressed TPA-induced phosphorylation and subsequent degradation of IkappaBalpha, and prevented nuclear translocation of p65. We also examined whether extracellular signal-regulated protein kinase (ERK) and p38 MAPK, which are known to regulate activation of NF-kappaB, can also modulate CREB DNA binding. Pretreatment with U0126 and SB203580, pharmacological inhibitors of ERK and p38 MAPK, respectively, showed that SB203580, but not U0126, attenuated TPA-induced CREB DNA binding in mouse skin. Taken together, EGCG inhibited TPA-induced DNA binding of NF-kappaB and CREB by blocking activation of p38 MAPK, which may provide a molecular basis of COX-2 inhibition by EGCG in mouse skin in vivo. PMID- 17404064 TI - Peonidin inhibits phorbol-ester-induced COX-2 expression and transformation in JB6 P+ cells by blocking phosphorylation of ERK-1 and -2. AB - Abnormal upregulation of cyclooxygenase-2 (COX-2) has been frequently observed in various types of transformed and cancerous cells. Numerous anti-inflammatory agents have been shown to exert chemopreventive effects by targeting COX-2, a rate-limiting enzyme involved in the inflammatory process. Anthocyanins are naturally occurring polyphenolic compounds that endow various fruits, vegetables, and plants with intense colors. Peonidin is another representative anthocyanidin, but its chemopreventive potential has not been fully described. This article investigated the effect of peonidin on 12-O-tetradecanoylphorbol-13-acetate (TPA) induced COX-2 expression and transformation in JB6 P(+) mouse epidermal cells (JB6 P(+) cells). Treatment of JB6 P(+) cells with peonidin inhibited TPA-induced COX-2 expression, and also decreased TPA-induced neoplastic transformation and blocked TPA-induced phosphorylation of extracellular signal-regulated kinases (ERKs) in the cells. The inhibition of the signaling mechanism regulating the activation of ERKs strongly suggests that peonidin exhibits chemopreventive as well as anti-inflammatory activities. PMID- 17404065 TI - Wogonin, a plant flavone, potentiates etoposide-induced apoptosis in cancer cells. AB - Etoposide, a podophylotoxin anticancer agent, induces apoptotic cell death in normal and cancer cells. Etoposide-induced apoptosis plays a role in not only anticancer effect but also adverse reaction, such as myelosuppression. Since we have found that wogonin, a flavone found in Scutellaria baicalensis Georgi, prevents thymocyte apoptosis induced by various compounds including etoposide, we examined the effect of this flavone on etoposide-induced apoptosis in cancer cells. Although 100 muM wogonin itself significantly increased DNA fragmentation in HL-60 cells, this change was not observed in Jurkat cells. On the other hand, this flavone significantly potentiated etoposide-induced apoptosis in Jurkat and HL-60 cells. Similarly, wogonin accelerated etoposide-induced cell death in lung cancer cells. Since wogonin had no effect on the action of other anticancer agents, such as 5-FU and cisplatin, this flavone seems to accelerate only etoposide-induced apoptotic cell death in cancer cells. These results suggest that the modification of etoposide-induced apoptosis by wogonin may be available to reduce the adverse reaction of this agent. PMID- 17404066 TI - Inhibitory effects of 7-carboxymethyloxy-3',4',5-trimethoxyflavone (DA-6034) on Helicobacter pylori-induced NF-kappa B activation and iNOS expression in AGS cells. AB - The Helicobacter pylori were identified by Marshall and Warren in 1984. H. pylori survive in the forbidding harsh acid environment of the stomach and duodenum by hiding in the mucus layer and neutralizing gastric acid in its local surrounding environment. Multiple lines of evidence suggest that H. pylori infection is one of the primary causes of gastritis and peptic ulcer, which are provoked by oxidative stress and inflammation. More than 50% of the world's population is infected by this bacterium. The H. pylori-induced inflammation has been implicated in the pathogenesis and progression of gastric cancer. DA-6034 (7 carboxymethyloxy-3',4',5-trimethoxy flavone) is a synthetic flavonoid known to possess anti-inflammatory activity. It has been reported that oral administration of DA-6034 suppresses the inflammatory bowel disease (IBD) in animal models. In this article, we attempted to examine the effect of DA-6034 on H. pylori-induced inflammation in human gastric cancer (AGS) cells by targeting NF-kappaB and extracellular signal-regulated kinase (ERK), a representative MAPK. PMID- 17404067 TI - Phenolic phytochemicals derived from red pine (Pinus densiflora) inhibit the invasion and migration of SK-Hep-1 human hepatocellular carcinoma cells. AB - Considerable attention has recently been focused on identifying chemopreventive phytochemicals derived from medicinal plants. Here, we analyzed phenolic phytochemicals from red pine (RP) leaves and found epigallocatechin gallate (EGCG) and epigallocatechin (EGC), and catechin gallate (CG) as their major phenolic phytochemicals. This article also investigated whether RP leaf extract and its phenolic phytochemicals inhibit the invasion of SK-Hep-1 human hepatocellular carcinoma cells (SK-Hep-1 cells). RP suppressed the invasion and the migration of SK-Hep-1 cells. EGCG and CG also inhibited the invasion and migration, with EGC exhibiting a lower efficacy. Matrix metalloproteinases (MMPs), particularly gelatinase-A (MMP-2) and gelatinase-B (MMP-9), degrade components of the basement membrane and are strongly implicated in invasion and metastasis formation of malignant tumors. RP suppressed both MMP-2 and MMP-9 activities. EGCG and CG reduced the activities of MMP-9 and MMP-2 in a dose dependent manner, with EGC exhibiting a lower efficacy on both MMPs. Our results suggest that RP inhibits tumor invasion and migration, which may be attributed to the effects of EGCG and CG. In particular, EGCG plays a key role in the efficacy of RP against hepatocarcinogenesis. PMID- 17404068 TI - KG-135 inhibits COX-2 expression by blocking the activation of JNK and AP-1 in phorbol ester-stimulated human breast epithelial cells. AB - Ginsenosides, ingredients of ginseng, have a wide array of pharmacologic effects. Especially, ginsenosides Rk1, Rg3, and Rg5 derived from heat-processed ginseng have been shown to possess substantial anti-tumor-promoting effects. KG-135 is a formulated complex that contains several antitumorigenic ginsenosides, such as Rk1, Rg3, Rg5, Rk2, Rk3, Rs3, Rs4, Rs5, Rs6, Rs7, etc. The present article was aimed at evaluating the chemopreventive as well as anti-inflammatory effects of KG-135 in the human breast epithelial cell line (MCF-10A). One of the well recognized molecular targets for chemoprevention is cyclooxygenase-2 (COX-2) that is abnormally upregulated in many premalignant and malignant tissues and cells. In this study, we found that KG-135 inhibited COX-2 expression in MCF-10A cells stimulated with a prototype tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA). Since the transcription factor activator protein-1 (AP-1) plays a role in tumor promotion and is also known to regulate COX-2 induction, we attempted to determine the effect of KG-135 on TPA-induced activation of AP-1. Cotreatment with KG-135 resulted in a decrease in TPA-induced DNA binding of AP-1. In addition, KG-135 inhibited TPA-induced phosphorylation of c-Jun N-terminal kinases (JNK) that regulates COX-2 expression in MCF-10A cells. The JNK inhibitor SP600125 attenuated COX-2 expression in TPA-treated MCF-10A cells. Taken together, the above findings suggest that KG-135 inhibits TPA-induced COX-2 expression in MCF-10A cells by blocking the JNK/AP-1 signaling pathway. PMID- 17404069 TI - Resveratrol inhibits IL-1 beta-induced stimulation of caspase-3 and cleavage of PARP in human articular chondrocytes in vitro. AB - Resveratrol is a polyphenolic phytoalexin that is present in various fruits, in the skin of red grapes and peanuts. Recent studies have shown that resveratrol exhibits potent antioxidant properties and is able to exert anti-inflammatory and anti-catabolic properties in several cell types. The pro-inflammatory cytokine interleukin-1beta (IL-1beta) plays a pivotal role in the pathogenesis of osteoarthritis (OA) in humans and animals. In this article we investigated whether resveratrol is able to block the effects of IL-1beta, specifically the activation of caspase-3 and subsequent cleavage of poly (ADP-ribose) polymerase (PARP) in human articular chondrocytes. Cultures of human chondrocytes were prestimulated with 10 ng/mL IL-1beta for 1, 12, and 24 h before being co-treated with IL-1beta and 100 microM resveratrol or 50 microM of the caspase inhibitor Z DEVD-FMK for 1, 12, and 24 h, respectively in vitro. Resveratrol significantly reduced the IL-1beta-induced inhibition of expression of cartilage-specific collagen type II and signal transduction receptor beta1-integrin in a time dependent manner. Incubation of chondrocytes with IL-1beta resulted in the activation of caspase-3 and PARP cleavage. These effects were abolished through co-treatment with resveratrol. Furthermore, co-treatment of IL-1beta-stimulated cells with the caspase inhibitor Z-DEVD-FMK blocked activation of caspase-3 and PARP cleavage, suggesting that this process is a caspase-dependent pathway. In summary, our results confirm that resveratrol is an effective inhibitor of chondrocyte apoptosis in vitro. These findings suggest that this dietary polyphenolic compound may have future applications in the nutraceutical-based therapy of human and animal OA. PMID- 17404070 TI - Possible link between NO concentrations and COX-2 expression in systems treated with soy-isoflavones. AB - The production of nitric oxide (NO) emerges as an essential determinant in auto- and paracrine signaling. NO is known to be generated under inflammatory conditions, carcinogenesis, and circulatory shock. The large amount of NO produced in response to cytokines plays an important role in inflammatory conditions. Cyclooxygenase (COX), the central enzyme in prostanoid biosynthesis, is involved in the first step of prostanoid synthesis from arachidonic acid. The reported studies to evaluate the relationship between NO and COX-2 have revealed both inhibitory and stimulatory effects of NO on COX-2 expression. Genistein, one of soy-isoflavones, is a polyphenolic flavonoid and a potent antioxidant and anti inflammatory agent. In the present article, the effect of soy-isoflavones on NO production and COX-2 gene expression was examined. NO production by soy isoflavones was greatly increased even though eNOS and iNOS expression were not different from nontreated control. The increment of NO was accompanied with the elevated expression of COX-2 and the concentrations of PGE2. The COX-2 stimulatory effect of soy-isoflavones appeared to be modulated by ERK-1 and -2 and p38. In mammalian cancer system, incubation with the NO donor sodium nitroprusside (SNP) resulted in a slight upregulation of COX-2, and cotreatment with genistein decreased COX-2 expression possibly by the activation of AMP activated protein kinase (AMPK). PMID- 17404071 TI - Assessment of the effect of Echinacea purpurea (L.) Moench on apoptotic and mitotic activity of liver cells during intoxication by cadmium. AB - Cadmium (Cd(2+)) is an important industrial pollutant, although its mechanism of toxicity has not been completely clarified. Cd(2+) is toxic to a wide range of organs and tissues, however, the primary target organs of Cd(2+) toxicity are the liver and kidney. Echinacea purpurea stimulating one or another tread of the immune system stimulates the expression of immunoglobulins and interferons. The experiments were performed on white laboratory mice using intraperitoneal (i.p.) injections 0.05 LD(50) amount of CdCl(2) solution. Two groups of mice were injected by Echinacea purpurea liquid extract: one 0.05 LD(50) and the other 0.1 LD(50). In this article, the Cd(2+) distribution in internal organs, its effect on the mitotic and apoptotic activity of liver cells, as well as effects of Echinacea purpurea liquid extract on Cd(2+)-induced changes in mice were investigated. Cd(2+) concentration in mice blood, liver, and kidney was detected by atomic absorption spectroscopy. Long-term injections of extract of Echinacea purpurea combined with Cd(2+)Cl(2) leads to the significant increase of Cd(2+) concentration in blood and investigated organs of experimental mice. Mitotic and apoptotic activity of liver cells was expressed as the estimated number of mitotic and apoptotic liver cells in randomly selected reference areas in histological slide. Echinacea purpurea decreases the mitotic activity of liver cells induced by Cd(2+) and increases apoptotic activity of the liver cells. PMID- 17404072 TI - Influence of Echinacea purpurea (L.) Moench extract on the toxicity of cadmium. AB - Echinacea purpurea (L.) Moench (EP) has many beneficial features, especially strengthening the immune system. Cadmium (Cd) is a ubiquitous metal and cumulative poison that may cause liver and kidney damage and the formation of neoplasia. In this article, the changes in organs and metabolism, the accumulation of various levels of Cd in tissues, and the effects of EP liquid extract on Cd-induced changes in mice were investigated. Experiments were carried out on the white laboratory mice. Solution of different Cd and EP concentrations were given to drink and experiments were performed. The concentration of Cd in mice blood, liver, kidneys, heart, spleen, skeletal muscle was determined using an electrothermal graphite furnace atomic absorption spectrophotometer Perkin Elmer/Zeeman 3030. Long time per os of extract of EP combined with Cd leads to a significant increase of Cd concentration in blood and investigated organs of experiment mice. PMID- 17404073 TI - Hypoxia and metastasis. PMID- 17404074 TI - Unraveling the mechanisms of endocrine resistance in breast cancer: new therapeutic opportunities. AB - Two thirds of breast cancers express the estrogen receptor (ER), which contributes to tumor development and progression. ER-targeted therapy is therefore widely used in breast cancer to inhibit signaling through ER and disrupt breast cancer growth. This therapeutic strategy, particularly using the antiestrogen tamoxifen, is proven to increase the cure rates in early breast cancer, improve patient outcomes in advanced disease, and reduce breast cancer incidence in the prevention setting. Despite the recent integration of more powerful endocrine agents into breast cancer care, resistance to all forms of endocrine therapy remains a major problem. New insight into ER biology and progress in understanding resistance mechanisms, mediated by molecular crosstalk between ER and various growth factor signaling pathways, are generating tremendous promise for new therapeutic opportunities to target resistance and improve breast cancer disease outcomes. PMID- 17404075 TI - Targeting checkpoint kinase 1 in cancer therapeutics. AB - Progression through the cell cycle is monitored by surveillance mechanisms known as cell cycle checkpoints. Our knowledge of the biochemical nature of checkpoint regulation during an unperturbed cell cycle and following DNA damage has expanded tremendously over the past decade. We now know that dysfunction in cell cycle checkpoints leads to genomic instability and contributes to tumor progression, and most agents used for cancer therapy, such as cytotoxic chemotherapy and ionizing radiation, also activate cell cycle checkpoints. Understanding how checkpoints are regulated is therefore important from the points of view of both tumorigenesis and cancer treatment. In this review, we present an overview of the molecular hierarchy of the checkpoint signaling network and the emerging role of checkpoint targets, especially checkpoint kinase 1, in cancer therapy. Further, we discuss the results of recent clinical trials involving the nonspecific checkpoint kinase 1 inhibitor, UCN-01, and the challenges we face with this new therapeutic approach. PMID- 17404076 TI - The potential of antiangiogenic therapy in non-small cell lung cancer. AB - The long-term prognosis for patients with advanced non-small cell lung cancer (NSCLC) remains poor despite the availability of several cytotoxic chemotherapy regimens. The use of targeted therapies, particularly those against the key mediator of angiogenesis vascular endothelial growth factor (VEGF), has the potential to improve outcomes for NSCLC patients. Bevacizumab, a recombinant humanized monoclonal anti-VEGF antibody, is the most clinically advanced antiangiogenic agent in NSCLC. In a phase III study, bevacizumab showed significantly improved overall and progression-free survival when used in combination with standard first-line chemotherapy in patients with advanced NSCLC. Bevacizumab was generally well tolerated in patients with NSCLC; however, tumor-related bleeding adverse events have been noted in some patients, predominantly those with squamous cell histology or centrally located tumors. Several small-molecule VEGF receptor tyrosine kinase inhibitors have also shown promise in phase I and II trials in NSCLC. This review summarizes the most important findings of angiogenesis inhibitors in NSCLC and discusses the potential for the use of these novel agents in different settings of NSCLC. PMID- 17404077 TI - Beta-2-microglobulin is an androgen-regulated secreted protein elevated in serum of patients with advanced prostate cancer. AB - PURPOSE: A better understanding of secreted proteins may lead to the discovery of new biomarkers, which, along with prostate-specific antigen (PSA), may be useful in the diagnosis and treatment of prostate cancer patients. EXPERIMENTAL DESIGN: Conditioned medium was collected from LNCaP cells following stimulation with methyltrienolone (R1881), 17beta-estradiol (estradiol), or interleukin-6 and analyzed for differential protein expression with surface-enhanced laser desorption/ionization-time of flight mass spectrometry. Quantitative reverse transcription-PCR, immunoblots, and ELISA were used to measure beta-2 microglobulin (B2M) message and protein levels in cells, conditioned medium, and serum. RESULTS: Surface-enhanced laser desorption/ionization-time of flight revealed that many peaks were induced or repressed following stimulation with R1881 or estradiol. A peak of interest centered at 11.8 kDa was chosen for additional analysis. Immunodepletion identified the peak of interest as B2M. Reverse transcription-PCR and immunoblots confirmed that PSA and B2M were induced by R1881. However, unlike PSA, B2M was not increased on stimulation with estradiol or interleukin-6. Human B2M is identified in the serum of mice bearing human prostate cancer xenograft. B2M is expressed in human prostate cancer cell lines and tissues. Serum B2M levels are elevated in patients with metastatic, androgen-independent prostate cancer. CONCLUSIONS: B2M is a secreted protein expressed in prostate cancer, which is more specific for androgen stimulation than PSA under the conditions tested. Additional studies are warranted to explore if B2M is as useful marker for prostate cancer. Identification of proteins secreted from cancer cells in preclinical models may be a useful strategy for biomarker discovery. PMID- 17404078 TI - Estrogen receptor beta expression is associated with tamoxifen response in ERalpha-negative breast carcinoma. AB - PURPOSE: Endocrine therapies, such as tamoxifen, are commonly given to most patients with estrogen receptor (ERalpha)-positive breast carcinoma but are not indicated for persons with ERalpha-negative cancer. The factors responsible for response to tamoxifen in 5% to 10% of patients with ERalpha-negative tumors are not clear. The aim of the present study was to elucidate the biology and prognostic role of the second ER, ERbeta, in patients treated with adjuvant tamoxifen. EXPERIMENTAL DESIGN: We investigated ERbeta by immunohistochemistry in 353 stage II primary breast tumors from patients treated with 2 years adjuvant tamoxifen, and generated gene expression profiles for a representative subset of 88 tumors. RESULTS: ERbeta was associated with increased survival (distant disease-free survival, P = 0.01; overall survival, P = 0.22), and in particular within ERalpha-negative patients (P = 0.003; P = 0.04), but not in the ERalpha positive subgroup (P = 0.49; P = 0.88). Lack of ERbeta conferred early relapse (hazard ratio, 14; 95% confidence interval, 1.8-106; P = 0.01) within the ERalpha negative subgroup even after adjustment for other markers. ERalpha was an independent marker only within the ERbeta-negative tumors (hazard ratio, 0.44; 95% confidence interval, 0.21-0.89; P = 0.02). An ERbeta gene expression profile was identified and was markedly different from the ERalpha signature. CONCLUSION: Expression of ERbeta is an independent marker for favorable prognosis after adjuvant tamoxifen treatment in ERalpha-negative breast cancer patients and involves a gene expression program distinct from ERalpha. These results may be highly clinically significant, because in the United States alone, approximately 10,000 women are diagnosed annually with ERalpha-negative/ERbeta-positive breast carcinoma and may benefit from adjuvant tamoxifen. PMID- 17404079 TI - 3q26 Amplification and polysomy of chromosome 3 in squamous cell lesions of the lung: a fluorescence in situ hybridization study. AB - PURPOSE: An overlapping area of gain at 3q26 has been reported in lung squamous cell carcinoma (SCC), but whether this also occurs in preneoplastic/preinvasive squamous cell proliferations and early-stage invasive carcinomas of the lung is still unknown. EXPERIMENTAL DESIGN: We evaluated the prevalence and the clinicopathologic implications of 3q26 amplification and polysomy of chromosome 3 in 31 preneoplastic/preinvasive squamous cell lesions of the bronchial mucosa and in 139 early-stage invasive pulmonary SCC, both of limited growth within the bronchial wall [early hilar SCC (EHSCC)] and involving the pulmonary parenchyma [parenchyma-infiltrating SCC (PISCC)]. Moreover, mRNA expression of two candidate genes (h-TERC and SKI-like), both mapping to the minimal common amplification region, was also studied by quantitative real-time reverse transcription-PCR. RESULTS: 3q26 amplification and polysomy of chromosome 3 were confined to malignant samples, with 37% of invasive SCC, and 27% of severe dysplasias/in situ carcinomas showing these chromosomal abnormalities. Amplification (with minimal common amplification region at 3q26.2), polysomy 3, concurrent amplification and polysomy 3, or other changes (monosomy) were found in 25 SCC and 1 dysplasia, 24 and 2, 2 and 0, and 1 and 0, respectively. Amplification was significantly associated with EHSCC, polysomy 3 with PISCC. 3q26 amplification correlated with increased tumor diameter and a history of smoking, whereas polysomy 3 correlated with tumor diameter, pT class, and p53, p21, and fascin immunoreactivity. No relationship of either 3q26 gain or polysomy was found with patients' survival. Overexpression of h-TERC or SKI-like mRNA was found in 3q26-amplified or polysomic SCC, with higher levels of h-TERC in the former and of SKI-like in the latter. CONCLUSIONS: 3q26 amplification and chromosome 3 polysomy may be related to the development of invasive SCC, with differential distribution in tumor subsets, despite substantial histologic uniformity. Both h-TERC and SKI-like may be involved in tumor progression. PMID- 17404080 TI - Molecular characterization of Waldenstrom's macroglobulinemia reveals frequent occurrence of two B-cell clones having distinct IgH VDJ sequences. AB - PURPOSE: Malignant B lineage cells in Waldenstrom's macroglobulinemia (WM) express a unique clonotypic IgM VDJ. The occurrence of biclonal B cells and their clonal relationships were characterized. EXPERIMENTAL DESIGN: Bone marrow and blood from 20 WM patients were analyzed for clonotypic VDJ sequences, clonal B cell frequencies, and the complementary determining region 3 profile. RESULTS: Two different clonotypic VDJ sequences were identified in 4 of 20 WM. In two cases, partner clones had different VDJ rearrangements, with one clonotypic signature in bone marrow and a second in blood. For both cases, the bone marrow clone was hypermutated, whereas the blood clone was germ line or minimally mutated. In two other cases, partner clones shared a common VDJ rearrangement but had different patterns of somatic mutations. They lacked intraclonal diversity and were more abundant in bone marrow than in blood. VDJ mutation profiles suggested they arose from a common IgM progenitor. Single-cell analysis in one case indicated the partner clones were reciprocally expressed, following rules of allelic exclusion. CONCLUSIONS: The existence of two B-cell clones having distinct VDJ sequences is common in WM, suggesting that frequent transformation events may occur. In two cases, the partner clones had distinct tissue distributions in either blood or bone marrow, were of different immunoglobulin isotypes, and in one case exhibited differential response to therapy. The contributions of each clone are unknown. Their presence suggests that WM may involve a background of molecular and cellular events leading to emergence of one or more malignant clones. PMID- 17404081 TI - Population-based molecular prognosis of breast cancer by transcriptional profiling. AB - PURPOSE: The purpose of this study is to predict breast cancer recurrence and metastases and to identify gene signatures indicative of clinicopathologic characteristics using gene expression patterns derived from cDNA microarray. EXPERIMENTAL DESIGN: Expression profiles of 7,650 genes were investigated on an unselected group of 99 node-negative and node-positive breast cancer patients to identify prognostic gene signature of recurrence and metastases. The identified gene signature was validated on independent 78 patients with primary invasive carcinoma (T(1)/T(2) and N(0)) and on 58 patients with locally advanced breast cancer (T(3)/T(4) and/or N(2)). The gene predictors were identified using a combination of random forests and linear discriminant analysis function. RESULTS: This study identified a new 28-gene signature that achieved highly accurate disease-free survival and overall survival (both at P < 0.001, time-dependent receiver operating characteristic analysis) in individual breast cancer patients. Patients categorized into high-risk, intermediate-risk, and low-risk groups had distinct disease-free survival (P < 0.005, Kaplan-Meier analysis, log-rank test) in three patient cohorts. A strong association (P < 0.05) was identified between risk groups and tumor size, tumor grade, estrogen receptor and progesterone receptor status, and HER2/neu overexpression in the studied cohorts. We also identified 14-gene predictors of nodal status and 9-gene predictors of tumor grade. CONCLUSIONS: This study has established a population-based approach to predicting breast cancer outcomes at the individual level exclusively based on gene expression patterns. The 28-gene recurrence signature has been validated as quantifying the probability of recurrence and metastases in patients with heterogeneous histology and disease stage. PMID- 17404082 TI - Circulating tumor cell analysis in patients with progressive castration-resistant prostate cancer. AB - PURPOSE: To better direct targeted therapies to the patients with tumors that express the target, there is an urgent need for blood-based assays that provide expression information on a consistent basis in real time with minimal patient discomfort. We aimed to use immunomagnetic-capture technology to isolate and analyze circulating tumor cells (CTC) from small volumes of peripheral blood of patients with advanced prostate cancer. EXPERIMENTAL DESIGN: Blood was collected from 63 patients with metastatic prostate cancer. CTCs were isolated by the Cell Search system, which uses antibodies to epithelial cell adhesion marker and immunomagnetic capture. CTCs were defined as nucleated cells positive for cytokeratins and negative for CD45. Captured cells were analyzed by immunofluorescence, Papanicolau staining, and fluorescence in situ hybridization. RESULTS: Most patients (65%) had 5 or more CTCs per 7.5 mL blood sample. Cell counts were consistent between laboratories (c = 0.99) and did not change significantly over 72 or 96 h of storage before processing (c = 0.99). Their identity as prostate cancer cells was confirmed by conventional cytologic analysis. Molecular profiling, including analysis of epidermal growth factor receptor (EGFR) expression, chromosome ploidy, and androgen receptor (AR) gene amplification, was possible for all prostate cancer patients with >or=5 CTCs. CONCLUSIONS: The analysis of cancer-related alterations at the DNA and protein level from CTCs is feasible in a hospital-based clinical laboratory. The alterations observed in EGFR and AR suggest that the methodology may have a role in clinical decision making. PMID- 17404083 TI - Adrenal androgen levels as predictors of outcome in prostate cancer patients treated with ketoconazole plus antiandrogen withdrawal: results from a cancer and leukemia group B study. AB - PURPOSE: Adrenal androgens activate the androgen receptor and stimulate prostate cancer growth. Ketoconazole is used as an inhibitor of adrenal androgen synthesis in men with androgen-independent prostate cancer. This study analyzes the relationship between pretreatment androgen levels and outcome following ketoconazole treatment. EXPERIMENTAL DESIGN: Baseline levels of three adrenal androgens (androstenedione, dehydroepiandrostenedione, and dehydroepiandrostenedione-sulfate) and testosterone were measured. Regression models (logistic and proportional hazard) were used to assess the prognostic significance of these levels in predicting overall survival and prostate-specific antigen (PSA) response defined by Consensus Criteria. RESULTS: In 103 patients with available levels, PSA response rate was 28% and median response duration was 4.8 months. The median baseline androstenedione level was 0.64 ng/mL and was 0.88 ng/mL versus 0.53 ng/mL for those with and without a PSA response, respectively (P = 0.034). In univariate analysis, elevation of baseline androstenedione levels was predictive of PSA response [odds ratio, 2.26; 95% confidence interval (95% CI), 1.03-4.96; P = 0.043]. In multivariate analysis, both the uppermost and the middle tertile of baseline androstenedione level were associated with an improved overall survival compared with those in the lower tertile (hazard ratio, 0.59; 95% CI, 0.36-0.98; P = 0.40; hazard ratio, 0.53; 95% CI, 0.32-0.90; P = 0.018, respectively). A linear correlation was observed among all androgen levels. CONCLUSIONS: Higher androstenedione levels predict likelihood of response to ketoconazole and improved survival compared with patients with lower levels. These data suggest that therapy with ketoconazole is less effective in patients with low levels of androgen at baseline. PMID- 17404084 TI - Loss of the mismatch repair protein MSH6 in human glioblastomas is associated with tumor progression during temozolomide treatment. AB - PURPOSE: Glioblastomas are treated by surgical resection followed by radiotherapy [X-ray therapy (XRT)] and the alkylating chemotherapeutic agent temozolomide. Recently, inactivating mutations in the mismatch repair gene MSH6 were identified in two glioblastomas recurrent post-temozolomide. Because mismatch repair pathway inactivation is a known mediator of alkylator resistance in vitro, these findings suggested that MSH6 inactivation was causally linked to these two recurrences. However, the extent of involvement of MSH6 in glioblastoma is unknown. We sought to determine the overall frequency and clinical relevance of MSH6 alterations in glioblastomas. EXPERIMENTAL DESIGN: The MSH6 gene was sequenced in 54 glioblastomas. MSH6 and O(6)-methylguanine methyltransferase (MGMT) immunohistochemistry was systematically scored in a panel of 46 clinically well characterized glioblastomas, and the corresponding patient response to treatment evaluated. RESULTS: MSH6 mutation was not observed in any pretreatment glioblastoma (0 of 40), whereas 3 of 14 recurrent cases had somatic mutations (P = 0.015). MSH6 protein expression was detected in all pretreatment (17 of 17) cases examined but, notably, expression was lost in 7 of 17 (41%) recurrences from matched post-XRT + temozolomide cases (P = 0.016). Loss of MSH6 was not associated with O(6)-methylguanine methyltransferase status. Measurements of in vivo tumor growth using three-dimensional reconstructed magnetic resonance imaging showed that MSH6-negative glioblastomas had a markedly increased rate of growth while under temozolomide treatment (3.17 versus 0.04 cc/mo for MSH6 positive tumors; P = 0.020). CONCLUSIONS: Loss of MSH6 occurs in a subset of post XRT + temozolomide glioblastoma recurrences and is associated with tumor progression during temozolomide treatment, mirroring the alkylator resistance conferred by MSH6 inactivation in vitro. MSH6 deficiency may therefore contribute to the emergence of recurrent glioblastomas during temozolomide treatment. PMID- 17404085 TI - Promoter hypermethylation identifies progression risk in bladder cancer. AB - PURPOSE: New methods to accurately predict an individual tumor behavior are urgently required to improve the treatment of cancer. We previously found that promoter hypermethylation can be an accurate predictor of bladder cancer progression, but it is not cancer specific. Here, we investigate a panel of methylated loci in a prospectively collected cohort of bladder tumors to determine whether hypermethylation has a useful role in the management of patients with bladder cancer. EXPERIMENTAL DESIGN: Quantitative methylation specific PCR was done at 17 gene promoters, suspected to be associated with tumor progression, in 96 malignant and 30 normal urothelial samples. Statistical analysis and artificial intelligence techniques were used to interrogate the results. RESULTS: Using log-rank analysis, five loci were associated with progression to more advanced disease (RASSF1a, E-cadherin, TNFSR25, EDNRB, and APC; P < 0.05). Multivariate analysis revealed that the overall degree of methylation was more significantly associated with subsequent progression and death (Cox, P = 0.002) than tumor stage (Cox, P = 0.008). Neuro-fuzzy modeling confirmed that these five loci were those most associated with tumor progression. Epigenetic predictive models developed using artificial intelligence techniques identified the presence and timing of tumor progression with 97% specificity and 75% sensitivity. CONCLUSION: Promoter hypermethylation seems a reliable predictor of tumor progression in bladder cancer. It is associated with aggressive tumors and could be used to identify patients with either superficial disease requiring radical treatment or a low progression risk suitable for less intensive surveillance. Multicenter studies are warranted to validate this marker. PMID- 17404086 TI - Plasma matrix metalloproteinase-9 level is better than serum matrix metalloproteinase-9 level to predict gastric cancer evolution. AB - PURPOSE: Matrix metalloproteinase-9 (MMP-9) in blood is a promising new tumor marker. The aims of the present study are to compare the usefulness of plasma and serum MMP-9 levels for predicting gastric cancer development, invasion, and survival. EXPERIMENTAL DESIGN: In this nested case-control study, 114 gastric cancer patients and 87 healthy controls were enrolled. MMP-9 levels and activities were quantitatively measured by ELISA assay and zymography. The results were compared with the occurrence, clinicopathologic features, and outcomes of gastric cancer patients. The follow-up time for all patients was at least 5 years. RESULTS: Serum MMP-9 levels were significantly higher than plasma MMP-9 levels. Both plasma and serum MMP-9 levels correlated significantly with active MMP-9 identified by zymography (P = 0.002 and P = 0.048, respectively). Plasma MMP-9 level was significantly elevated in gastric cancer patients when compared with control subjects (P < 0.001). Serum MMP-9 levels did not differ between the groups. Receiver-operator characteristics analysis showed the values of sensitivity (82.5%) and specificity (65.5%) at the maximum accuracy for plasma MMP-9 at >or=60 ng/mL (P < 0.001). Elevated plasma MMP-9 correlated significantly with lymph node metastasis [odds ratio (OR), 3.43; P = 0.019], lymphatic invasion (OR, 7.58; P = 0.009), and venous invasion (OR, 4.14; P = 0.033). Patients with elevated plasma MMP-9 levels had poorer survival rates than those with normal plasma MMP-9 levels (P = 0.038). Serum MMP-9 level did not correlate well with gastric cancer-invasive phenotypes or survival. CONCLUSION: Our results suggest plasma MMP-9 level is a better marker than serum MMP-9 level for predicting gastric cancer development and progression. PMID- 17404087 TI - Microtubule-associated protein-tau is a bifunctional predictor of endocrine sensitivity and chemotherapy resistance in estrogen receptor-positive breast cancer. AB - PURPOSE: The clinical outcome for patients with breast cancer is influenced by the metastatic competence of the cancer and its sensitivity to endocrine therapy and chemotherapy. A molecular marker may be prognostic of outcome or predictive of response to therapy, or a combination of both. EXPERIMENTAL DESIGN: We examined separately the prognostic and predictive values of tau mRNA expression in estrogen receptor (ER)-positive primary breast cancers in three patient cohorts. We used gene expression data from 209 untreated patients to assess the pure prognostic value of tau, data from 267 patients treated with adjuvant tamoxifen to assess predictive value for endocrine therapy, and data from 82 patients treated with preoperative paclitaxel followed by 5-fluorouracil, doxorubicin, and cyclophosphamide (paclitaxel/FAC) to assess predictive value for chemotherapy response. Wilcoxon rank sum test was used to compare tau expression between different outcome groups. RESULTS: Higher tau mRNA expression showed borderline nonsignificant association with better prognosis in the absence of systemic adjuvant therapy. Higher tau mRNA expression was significantly associated with no recurrence (at 5 and 10 years, P = 0.005 and P = 0.05, respectively) in patients treated with tamoxifen, indicating a predictive value for endocrine therapy. Tau expression was significantly lower in patients who achieved pathologic complete response to paclitaxel/FAC chemotherapy (P < 0.001). CONCLUSION: This study suggests that high tau mRNA expression in ER-positive breast cancer indicates an endocrine-sensitive but chemotherapy-resistant disease. In contrast, low tau expression identifies a subset of ER-positive cancers that have poor prognosis with tamoxifen alone and may benefit from taxane containing chemotherapy. PMID- 17404088 TI - Utility of circulating B-RAF DNA mutation in serum for monitoring melanoma patients receiving biochemotherapy. AB - PURPOSE: Somatic B-RAF gene mutation has been identified in many malignancies and detected at a high frequency in cutaneous malignant melanoma. However, the significance of the B-RAF mutation (B-RAFmt) in terms of its prognostic and predictive capabilities for treatment response or disease outcome is not known. We hypothesized that circulating serum B-RAFmt (B-RAFsmt) at V600E, detected in serum, predicts response in melanoma patients receiving concurrent biochemotherapy. EXPERIMENTAL DESIGN: A real-time clamp quantitative reverse transcription-PCR assay was designed to assess B-RAFsmt by peptide nucleic acid clamping and a locked nucleic acid hybrid probe. Normal (n = 18) and American Joint Committee on Cancer stage I to IV melanoma patients (n = 103) were evaluated. These included stage IV patients (n = 48) with blood drawn before and after biochemotherapy. Patients were classified as biochemotherapy responders or nonresponders. Responders (n = 24) had a complete or partial response following biochemotherapy; nonresponders (n = 24) developed progressive disease. RESULTS: Of the 103 melanoma patients, 38 (37%) had B-RAFsmt DNA, of which 11 of 34 (32%) were stage I or II, and 27 of 69 (39%) were stage III or IV. Of the 48 biochemotherapy patients, 10 of 24 (42%) patients were positive for the B-RAFsmt in the respective responder and nonresponder groups before treatment. After biochemotherapy, B-RAFsmt was detected in only 1 of 10 patients (10%) in the responder group and 7 of 10 patients (70%) in the nonresponder group. B-RAFsmt is associated with significantly worse (P = 0.039) overall survival in patients receiving biochemotherapy. CONCLUSION: These studies show the presence and utility of circulating B-RAFsmt DNA in melanoma patients. PMID- 17404089 TI - Tumor-infiltrating Foxp3-CD4+CD25+ T cells predict poor survival in renal cell carcinoma. AB - PURPOSE: Regulatory T cells (Tregs) have been implicated as inhibitors of antitumoral immunity, and evidence suggests that elimination of Tregs may augment natural and pharmacologic immunity. We tested for the presence of putative Tregs within renal cell carcinoma (RCC) tumors. EXPERIMENTAL DESIGN: We identified 170 patients who underwent radical or partial nephrectomy for clear cell RCC between 2000 and 2002. Specimens were stained with anti-CD4, anti-CD25, and anti-Foxp3 antibodies and examined using confocal microscopy. Associations of CD4(+)CD25(+)Foxp3(-) and CD4(+)CD25(+)Foxp3(+) T cells with death from RCC were evaluated using Cox proportional hazards regression models. RESULTS: At last follow-up, 46 of 170 patients had died; of these, 37 died from RCC at a median of 1.4 years following nephrectomy (range, 0-4.4). Among the 124 remaining patients, median follow-up was 3.7 years (range, 0-5.7). Forty-three (25.3%) tumors harbored CD4(+)CD25(+)Foxp3(+) T cells. The presence of Foxp3(+) T cells was not significantly associated with RCC death univariately. One hundred forty-three (84.1%) tumors harbored CD4(+)CD25(+)Foxp3(-) T cells. The indicator for >or=10% CD4(+)CD25(+)Foxp3(-) T cells was significantly associated with RCC death univariately [risk ratio (RR), 2.60; 95% confidence interval (95% CI), 1.35-4.98; P = 0.004], after adjusting for tumor B7-H1 expression (RR, 2.53; 95% CI, 1.32 4.85; P = 0.005) and lymphocytic infiltration (RR, 2.53; 95% CI, 1.32-4.87; P = 0.005). CONCLUSIONS: Increased presence of CD4(+)CD25(+)Foxp3(+) T cells was not significantly associated with RCC death. In contrast, CD4(+)CD25(+)Foxp3(-) T cells, which may represent a unique set of Tregs or activated helper T cells, was significantly associated with outcome. PMID- 17404090 TI - Stromal myofibroblasts predict disease recurrence for colorectal cancer. AB - PURPOSE: Myofibroblasts, which are specifically differentiated fibroblasts, are thought to play a central role in the desmoplastic reaction, a dynamic stromal change closely associated with cancer development. Although fundamental studies suggest that myofibroblasts may either facilitate or inhibit cancer progression, cumulative evidence supports their role in promoting tumor progression. The aim of this study was to assess the value of myofibroblasts in the cancer stroma as an indicator of disease recurrence after colorectal cancer surgery. EXPERIMENTAL DESIGN: Using computer-assisted image analysis, we quantified myofibroblasts in the cancer-associated stroma of 192 colorectal cancers using alpha-smooth muscle actin as a marker. RESULTS: The cancer-associated stroma contained various numbers of myofibroblasts (0.35-19.0%; mean, 5.55 +/- 3.85%). Tumors with abundant myofibroblasts were associated with shorter disease-free survival rate (P = 0.001) for stage II and III colorectal cancer. Multivariate analysis indicated that alpha-smooth muscle actin was a significant prognostic factor comparable with lymph node metastasis and superior to other tumor and stromal components, including histology of the tumor invasive front, peritumoral lymphocytic infiltration, and Crohn's-like lymphoid reaction. Moreover, colorectal cancers with synchronous liver metastasis generally displayed an active desmoplastic reaction, which was retained in the metastatic lesion to a similar extent. CONCLUSIONS: The results suggest that the abundance of myofibroblasts in cancer-associated stroma may be a useful indicator of disease recurrence after curative colorectal cancer surgery. PMID- 17404091 TI - A phase I pharmacokinetic and pharmacodynamic study of s-3304, a novel matrix metalloproteinase inhibitor, in patients with advanced and refractory solid tumors. AB - PURPOSE: Matrix metalloproteinases (MMP) play a fundamental role in cancer development and progression. S-3304 is a potent, orally active, noncytotoxic inhibitor of MMPs, primarily MMP-2 and MMP-9, that prolongs survival in mice xenografts and is well tolerated in healthy volunteers. EXPERIMENTAL DESIGN: The aims of this phase I clinical trial were to determine the maximum tolerated dose, dose-limiting toxicities, pharmacokinetic profile, and intratumoral MMP inhibitory activity of single-agent S-3304 in advanced and refractory solid tumors. MMP activity was determined by film in situ zymography (FIZ). Patients had tumor biopsies before and after S-3304 administration and were also evaluated for response and survival. RESULTS: Four dose levels were explored [DL1-DL4 or 800, 1,600, 2,400, and 3200 mg twice daily (BID), respectively], and 32 patients were enrolled. Toxicities were mostly gastrointestinal. The maximum tolerated dose was not reached, but dose escalations beyond DL4 were impractical (number of capsules needed). S-3304 steady-state concentrations were reached by day 8, and day 1 mean C(max) and AUC(0-8) increases were less than dose proportional. After S-3304 administration, 17 of 18 patients experienced inhibition of MMP activity by FIZ. Strong mean inhibition of MMP activity was observed in DL1 to DL3. The negative mean inhibitory activity calculated for DL4 was due to one patient with a 397% MMP activity increase. CONCLUSION: S-3304 is safe, well tolerated, and achieves plasma concentrations above those required to inhibit MMP-2 and MMP-9. Its intratumoral MMP inhibitory activity has been shown using FIZ, which is useful as a biomarker with this and other MMP inhibitors. PMID- 17404092 TI - Effects of the administration of high-dose interleukin-2 on immunoregulatory cell subsets in patients with advanced melanoma and renal cell cancer. AB - PURPOSE: High-dose recombinant human interleukin-2 (IL-2) therapy is of clinical benefit in a subset of patients with advanced melanoma and renal cell cancer. Although IL-2 is well known as a T-cell growth factor, its potential in vivo effects on human immunoregulatory cell subsets are largely unexplored. EXPERIMENTAL DESIGN: Here, we studied the effects of high-dose IL-2 therapy on circulating dendritic cell subsets (DC), CD1d-reactive invariant natural killer T cells (iNKT), and CD4(+)CD25(+) regulatory-type T cells. RESULTS: The frequency of both circulating myeloid DC1 and plasmacytoid DC decreased during high-dose IL 2 treatment. Of these, only a significant fraction of myeloid DC expressed CD1d. Although the proportion of Th1-type CD4(-) iNKT increased, similarly to DC subsets, the total frequency of iNKT decreased during high-dose IL-2 treatment. In contrast, the frequency of CD4(+)CD25(+) T cells, including CD4(+)Foxp3(+) T cells, which have been reported to suppress antitumor immune responses, increased during high-dose IL-2 therapy. However, there was little, if any, change of expression of GITR, CD30, or CTLA-4 on CD4(+)CD25(+) T cells in response to IL-2. Functionally, patient CD25(+) T cells at their peak level (immediately after the first cycle of high-dose IL-2) were less suppressive than healthy donor CD25(+) T cells and mostly failed to Th2 polarize iNKT. CONCLUSIONS: Our data show that there are reciprocal quantitative and qualitative alterations of immunoregulatory cell subsets with opposing functions during treatment with high-dose IL-2, some of which may compromise the establishment of effective antitumor immune responses. PMID- 17404093 TI - A Phase I pharmacokinetic and pharmacodynamic study of PX-12, a novel inhibitor of thioredoxin-1, in patients with advanced solid tumors. AB - PURPOSE: Thioredoxin-1 (Trx-1) is a cellular redox protein that promotes tumor growth, inhibits apoptosis, and up-regulates hypoxia-inducible factor-1alpha and vascular endothelial growth factor. Objectives of this study were to determine safety, tolerability, pharmacodynamics, and pharmacokinetics of PX-12, a small molecule inhibitor of Trx-1. EXPERIMENTAL DESIGN: Thirty-eight patients with advanced solid tumors received PX-12 at doses of 9 to 300 mg/m(2), as a 1- or 3-h i.v. infusion on days 1 to 5, repeated every 3 weeks. RESULTS: At the 300 mg/m(2) dose level, one patient experienced a reversible episode of pneumonitis during the first cycle, and a second patient developed pneumonitis after the second cycle. Doses up to 226 mg/m(2) were well tolerated, and grade 3/4 events were uncommon (<3% of patients). The limiting factor on this dosing schedule was pungent odor caused by expired drug metabolite, 2-butanethiol. The best response was stable disease in seven patients (126-332 days). Whereas PX-12 was not detectable following the infusion, the C(max) of its inactive metabolite, 2 mercaptoimidazole, increased linearly with dose. PX-12 treatment lowered plasma Trx-1 concentrations in a dose-dependent manner. CONCLUSIONS: PX-12, the first Trx-1 inhibitor to enter clinical trials, was tolerated up to a dose of 226 mg/m(2) by a 3-h infusion. Based on pharmacodynamic and pharmacokinetic data, a trial of prolonged infusion schedule of PX-12 has been initiated. PMID- 17404094 TI - Angiopoietin 2 is a potential mediator of high-dose interleukin 2-induced vascular leak. AB - PURPOSE: High-dose interleukin 2 (HDIL2) produces durable tumor regressions in 10% of patients with metastatic renal cell carcinoma and melanoma. However, a major toxicity is vascular leak syndrome (VLS). We previously reported elevated serum angiopoietin 2 (Ang2) in septic patients with vascular leak and hypothesized that Ang2 might also contribute to HDIL2 VLS. EXPERIMENTAL DESIGN: Blood was collected from 14 patients receiving HDIL2 and from 4 patients receiving HDIL2 and bevacizumab, an antibody against vascular endothelial growth factor (VEGF). The effect of Ang2 was studied in vitro by incubating high Ang2 patient serum with cultured endothelial cells. RESULTS: Pretreatment Ang2 levels were in the reference range (median, 3.3 ng/mL) and rose with each day of IL-2 therapy (median peak, 29.7 ng/mL). No trend was seen in free VEGF levels during therapy. Patients treated with HDIL2 and bevacizumab all developed VLS and elevated Ang2. High Ang2 patient sera induced propermeability structural changes in endothelial cells, an effect reversed by blockade with the competitive ligand angiopoietin 1 (Ang1). CONCLUSIONS: Ang2 may be a mediator of HDIL2 VLS as evidenced by (a) an increase in Ang2 in all patients on HDIL2; (b) the effect of high Ang2 patient serum on cultured endothelial cells; (c) rescue of those structural changes by Ang1. The lack of correlation between VLS and serum VEGF levels in patients treated with HDIL2 alone or in combination with bevacizumab suggests that VEGF is not a major contributor to VLS or Ang2 release. These data suggest that the inhibition of Ang2 may mitigate VLS in patients receiving HDIL2. PMID- 17404095 TI - Evaluation of plasma insulin-like growth factor binding protein 2 and Her-2 extracellular domain as biomarkers for 17-allylamino-17-demethoxygeldanamycin treatment of adult patients with advanced solid tumors. AB - PURPOSE: Interaction of 17-allylamino-17-demethoxygeldanamycin (17-AAG) with heat shock protein 90 results in proteasomal degradation of many proteins, including Her-2-neu, with subsequent decreased expression of insulin-like growth factor binding protein-2 (IGFBP-2). Concentrations of both IGFBP-2 and Her-2 extracellular domain (Her-2 ECD) in sera of mice bearing BT474 human breast cancer xenografts decrease after 17-AAG treatment. We investigated whether this phenomenon occurred in patients. MATERIALS AND METHODS: Eight to 15 plasma samples were obtained between 0 and 72 h from 27 patients treated with single agent 17-AAG at doses between 10 and 307 mg/m(2) and 18 patients treated with 17 AAG at doses between 220 and 450 mg/m(2) combined with 70 to 75 mg/m(2) of docetaxel. Pretreatment plasma samples were also obtained from 12 healthy volunteers. Plasma IGFBP-2 and Her-2 ECD concentrations were quantitated by ELISA. RESULTS: Pretreatment plasma IGFBP-2 concentrations in patients (171 +/- 116 ng/mL) were 2-fold higher than those in healthy volunteers (85 +/- 44 ng/mL; P < 0.05). Following 17-AAG treatment, there were no consistent dose-dependent or time-dependent changes in plasma IGFBP-2 and Her-2 ECD concentrations. IGFBP-2 concentrations decreased by >or=40% in 8 patients, increased 2- to 5-fold in 8 patients, and remained essentially unchanged in 29 patients. Her-2 ECD concentrations decreased by >or=40% in 10 patients, increased 1.5- to 5-fold in 2 patients, and remained essentially unchanged in 25 patients. CONCLUSIONS: As previously reported, IGFBP-2 concentrations in plasma of cancer patients are significantly higher than those in healthy volunteers. In contrast to a mouse model, 17-AAG treatment was not consistently associated with decreases in IGFBP-2 or Her-2 ECD concentrations in patient plasma. PMID- 17404096 TI - Phase I evaluation of a fully human anti-alphav integrin monoclonal antibody (CNTO 95) in patients with advanced solid tumors. AB - PURPOSE: A fully human monoclonal antibody to anti-alpha(v) integrins (CNTO 95) has been shown to inhibit angiogenesis and tumor growth in preclinical studies. We assessed the safety and pharmacokinetics of CNTO 95 in patients with advanced refractory solid tumors. EXPERIMENTAL DESIGN: In this phase I trial, CNTO 95 (0.1, 0.3, 1.0, 3.0, and 10.0 mg/kg) was infused on days 0, 28, 35, and 42, and clinical assessments, dynamic contrast-enhanced magnetic resonance imaging (DCE MRI), and [(18)F]-2-fluorodeoxyglucose positron emission tomography (FDG-PET) were done. Patients achieving stable disease or better were eligible for extended dosing every 3 weeks for up to 12 months. RESULTS: Among the 24 enrolled patients, CNTO 95 was associated with one episode of grade III and four episodes of grade II infusion-related fever (all responded to acetaminophen). Of the six patients who received extended dosing, one patient (10.0 mg/kg), with cutaneous angiosarcoma, had a 9-month partial response. Pre- and post-treatment lesion biopsies confirmed tumor cell alpha(v) integrin expression, as well as CNTO 95 penetration of the tumor and localization to tumor cells in association with reduced bcl-2 expression. A lesion in one patient (10.0 mg/kg) with stable ovarian carcinosarcoma was no longer detectable by FDG-PET by day 49. Exposure to CNTO 95 seemed to increase in a greater-than-dose-proportional manner; dose dependent mean half-life ranged from 0.26 to 6.7 days. CONCLUSIONS: CNTO 95 was generally well tolerated. Six patients received extended therapy, including one patient with a prolonged response. Biopsy data confirmed tumor localization and pharmacodynamic activity. PMID- 17404097 TI - 5-aza-2'-deoxycytidine delays androgen-independent disease and improves survival in the transgenic adenocarcinoma of the mouse prostate mouse model of prostate cancer. AB - PURPOSE: We have previously shown that 5-aza-2'-deoxycytidine (5-aza) is an effective chemopreventive agent capable of preventing early disease progression in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model. The purpose of this study was to determine the effect of 5-aza on preexisting TRAMP prostate cancers and prevention of androgen-independent prostate cancer. EXPERIMENTAL DESIGN: TRAMP mice with established prostate cancers were treated with 5-aza, castration, castration + 5-aza, or vehicle control (PBS). One cohort of 22 mice per treatment was euthanized after 10 weeks of treatment, whereas a second cohort of 14 mice per group was followed until death to determine survival. Histologic sections of prostate, pelvic lymph nodes, lung, and liver were blinded and analyzed by a certified genitourinary pathologist (K.J.W.). RESULTS: Combined treatment (castration + 5-aza) provided significant survival benefits over either single treatment (combined versus castration P = 0.029, combined versus 5-aza P = 0.036). At 24 weeks of age, 86% of mice within the PBS cohort exhibited histologic evidence of prostate cancer, whereas only 47% of the combined cohort exhibited malignant disease (P < 0.0001). Additionally, whereas 43% of the PBS treatment group exhibited lymph node metastases, these were only observed in 21% of the combined treatment mice. CONCLUSIONS: This is the first study to examine the effect of 5-aza and combined castration + 5-aza on preexisting prostate cancer in an animal model. Based on these preclinical findings, we suggest that 5 aza treatment may prolong the time to an androgen-independent status and thus survival in a hormone-deprived setting in prostate cancer. PMID- 17404098 TI - Mcl-1 is a relevant therapeutic target in acute and chronic lymphoid malignancies: down-regulation enhances rituximab-mediated apoptosis and complement-dependent cytotoxicity. AB - PURPOSE: The antiapoptotic Bcl-2 family member protein Mcl-1 is dynamically regulated in transformed B-cells, has a short mRNA and protein half-life, and is rapidly processed during apoptosis. Multiple therapies cause down-regulation of Mcl-1 in chronic and acute lymphoid leukemia (CLL and ALL) cells. Mcl-1 has also been reported to mediate resistance to rituximab in CLL. We therefore investigated whether direct reduction of Mcl-1 was sufficient to induce apoptosis and increase sensitivity to rituximab. EXPERIMENTAL DESIGN: We used Mcl-1 specific small interfering RNA in ALL cell lines and tumor cells from CLL patients to block transcription of Mcl-1. RESULTS: We show that Mcl-1 down regulation alone is sufficient to promote mitochondrial membrane depolarization and apoptosis in ALL and CLL cells. Given the importance of rituximab in B-cell malignancies, we next assessed the influence of Mcl-1 down-regulation on antibody mediated killing. Mcl-1 down-regulation by small interfering RNA increased sensitivity to rituximab-mediated killing both by direct apoptosis and complement dependent cytotoxicity, but did not enhance antibody-dependent cellular cytotoxicity. CONCLUSIONS: These results show that Mcl-1 is a relevant therapeutic target for ALL and CLL, and its down-regulation has the potential to enhance the therapeutic effect of rituximab in CD20-bearing lymphoid cells. PMID- 17404099 TI - Clinical significance and therapeutic potential of the programmed death-1 ligand/programmed death-1 pathway in human pancreatic cancer. AB - PURPOSE: The programmed death-1 ligand/programmed death-1 (PD-L/PD-1) pathway has been recently suggested to play a pivotal role in the immune evasion of tumors from host immune system. In this study, we tried to reveal the clinical importance and therapeutic potential of the PD-L/PD-1 pathway in pancreatic cancer, which is one of the most aggressive and intractable malignant tumors. EXPERIMENTAL DESIGN: We used immunohistochemistry to investigate PD-L expression in 51 patients with pancreatic cancer who underwent surgery and explored the therapeutic efficacy of blocking the PD-L1/PD-1 pathway in murine pancreatic cancer in vivo. RESULTS: PD-L1-positive patients had a significantly poorer prognosis than the PD-L1-negative patients, whereas there was no significant correlation of tumor PD-L2 expression with patient survival. PD-L1 expression was inversely correlated with tumor-infiltrating T lymphocytes, particularly CD8(+) T cells. These clinical data have suggested that the PD-L1/PD-1 pathway may be a critical regulator in human pancreatic cancer. Monoclonal antibodies against PD L1 or PD-1 induced a substantial antitumor effect on murine pancreatic cancer in vivo. PD-L1 blockade promoted CD8(+) T-cell infiltration into the tumor and induced local immune activation. Furthermore, the combination of anti-PD-L1 monoclonal antibody and gemcitabine exhibited a significant synergistic effect on murine pancreatic cancer and resulted in complete response without overt toxicity. CONCLUSION: Our data suggest for the first time that PD-L1 status may be a new predictor of prognosis for patients with pancreatic cancer and provide the rationale for developing a novel therapy of targeting the PD-L/PD-1 pathway against this fatal disease. PMID- 17404100 TI - Systemic CTLA-4 blockade ameliorates glioma-induced changes to the CD4+ T cell compartment without affecting regulatory T-cell function. AB - PURPOSE: Patients with malignant glioma suffer global compromise of their cellular immunity, characterized by dramatic reductions in CD4(+) T cell numbers and function. We have previously shown that increased regulatory T cell (T(reg)) fractions in these patients explain T-cell functional deficits. Our murine glioma model recapitulates these findings. Here, we investigate the effects of systemic CTLA-4 blockade in this model. EXPERIMENTAL DESIGN: A monoclonal antibody (9H10) to CTLA-4 was employed against well-established glioma. Survival and risks for experimental allergic encephalomyelitis were assessed, as were CD4(+) T cell numbers and function in the peripheral blood, spleen, and cervical lymph nodes. The specific capacities for anti-CTLA-4 to modify the functions of regulatory versus CD4(+)CD25(-) responder T cells were evaluated. RESULTS: CTLA-4 blockade confers long-term survival in 80% of treated mice, without eliciting experimental allergic encephalomyelitis. Changes to the CD4 compartment were reversed, as anti CTLA-4 reestablishes normal CD4 counts and abrogates increases in CD4(+)CD25(+)Foxp3(+)GITR(+) regulatory T cell fraction observed in tumor-bearing mice. CD4(+) T-cell proliferative capacity is restored and the cervical lymph node antitumor response is enhanced. Treatment benefits are bestowed exclusively on the CD4(+)CD25(-) T cell population and not T(regs), as CD4(+)CD25(-) T cells from treated mice show improved proliferative responses and resistance to T(reg) mediated suppression, whereas T(regs) from the same mice remain anergic and exhibit no restriction of their suppressive capacity. CONCLUSIONS: CTLA-4 blockade is a rational means of reversing glioma-induced changes to the CD4 compartment and enhancing antitumor immunity. These benefits were attained through the conferment of resistance to T(reg)-mediated suppression, and not through direct effects on T(regs). PMID- 17404101 TI - Accelerated preclinical testing using transplanted tumors from genetically engineered mouse breast cancer models. AB - PURPOSE: The use of genetically engineered mouse (GEM) models for preclinical testing of anticancer therapies is hampered by variable tumor latency, incomplete penetrance, and complicated breeding schemes. Here, we describe and validate a transplantation strategy that circumvents some of these difficulties. EXPERIMENTAL DESIGN: Tumor fragments from tumor-bearing MMTV-PyMT or cell suspensions from MMTV-PyMT, -Her2/neu, -wnt1, -wnt1/p53(+/-), BRCA1/p53(+/-), and C3(1)T-Ag mice were transplanted into the mammary fat pad or s.c. into naive syngeneic or immunosuppressed mice. Tumor development was monitored and tissues were processed for histopathology and gene expression profiling. Metastasis was scored 60 days after the removal of transplanted tumors. RESULTS: PyMT tumor fragments and cell suspensions from anterior glands grew faster than posterior tumors in serial passages regardless of the site of implantation. Microarray analysis revealed genetic differences between these tumors. The transplantation was reproducible using anterior tumors from multiple GEM, and tumor growth rate correlated with the number of transplanted cells. Similar morphologic appearances were observed in original and transplanted tumors. Metastasis developed in >90% of mice transplanted with PyMT, 40% with BRCA1/p53(+/-) and wnt1/p53(+/-), and 15% with Her2/neu tumors. Expansion of PyMT and wnt1 tumors by serial transplantation for two passages did not lead to significant changes in gene expression. PyMT-transplanted tumors and anterior tumors of transgenic mice showed similar sensitivities to cyclophosphamide and paclitaxel. CONCLUSIONS: Transplantation of GEM tumors can provide a large cohort of mice bearing mammary tumors at the same stage of tumor development and with defined frequency of metastasis in a well-characterized molecular and genetic background. PMID- 17404102 TI - Transfection with anti-p65 intrabody suppresses invasion and angiogenesis in glioma cells by blocking nuclear factor-kappaB transcriptional activity. AB - PURPOSE: The strategy of intracellular antibodies to neutralize the function of target proteins has been widely developed for cancer research. This study used an intrabody against p65 subunit to prevent nuclear factor kappaB (NF-kappaB) transcriptional activity in glioma cells and to inhibit the expression of its target genes involved in the invasion and angiogenesis of human gliomas. EXPERIMENTAL DESIGN: A single-chain fragment of antibody variable region (scFv) against p65 was prepared using phage display technique. We then prepared an anti p65 intrabody construct (pFv/nu) by cloning the scFv-encoding sequence into the mammalian nuclear-targeting vector, pCMV/myc/nuc. RESULTS: p65 expression in human glioma cells (U251 and] U87) transfected with pFv/nu was significantly decreased. We showed that NF-kappaB nuclear translocation and its DNA binding activity were blocked via intrabody transfection in electrophoretic mobility shift assays and the inhibition of NF-kappaB activity in nucleus resulted in the decreasing expression and bioactivity of matrix metalloproteinase-9, urokinase type plasminogen activator receptor, urokinase-type plasminogen activator, and vascular endothelial growth factor. The intrabody transfected glioma cells showed a markedly lower level of invasion in Matrigel invasion assay. The capillary-like structure formation of endothelial cells was also repressed by coculture with the intrabody transfected glioma cells or exposure to their conditional medium. Intrabody transfection neither induced apoptosis nor altered cell proliferation in U251 and U87 cells as compared with the control vector pCMV/nu. After the injection of pFv/nu-transfected glioma cells, preestablished tumors were almost completely regressed when compared with mock, pCMV/nu, and pGFP/nu. CONCLUSION: Blocking NF-kappaB activity via the nuclear intrabody expression might be a potential approach for cancer therapy. PMID- 17404103 TI - The therapeutic potential of hepatocyte growth factor to sensitize ovarian cancer cells to cisplatin and paclitaxel in vivo. AB - PURPOSE: Advanced ovarian cancers are initially responsive to combinatorial chemotherapy with platinum drugs and taxanes but, in most cases, develop drug resistance. We recently showed that, in vitro, hepatocyte growth factor (HGF) enhances death of human ovarian cancer cell lines treated with cisplatin (CDDP) and paclitaxel. The present study addresses whether in vivo HGF makes ovarian carcinoma cells more responsive to these chemotherapeutics. EXPERIMENTAL DESIGN: Using Lentiviral vectors carrying the HGF transgene, we transduced SK-OV-3 and NIH:OVCAR-3 ovarian carcinoma cell lines to obtain stable autocrine and paracrine HGF receptor activation. In vitro, we assayed growth, motility, invasiveness, and the response to CDDP and paclitaxel of the HGF-secreting bulk unselected cell populations. In vivo, we tested the cytotoxic effects of the drugs versus s.c. tumors formed by the wild-type and HGF-secreting cells in immunocompromised mice. Tumor-bearing mice were treated with CDDP (i.p.) and paclitaxel (i.v.), combined in different schedules and doses. RESULTS: In vitro, HGF-secreting cells did not show altered proliferation rates and survival but were strongly sensitized to the death triggered by CDDP and paclitaxel, alone or in combination. In vivo, we found a therapeutic window in which autocrine/paracrine HGF made tumors sensitive to low doses of the drugs, which were ineffective on their own. CONCLUSIONS: These data provide the proof-of-concept that in vivo gene therapy with HGF might be competent in sensitizing ovarian cancer cells to conventional chemotherapy. PMID- 17404104 TI - Chemosensitization of cancer in vitro and in vivo by nitric oxide signaling. AB - PURPOSE: Hypoxia contributes to drug resistance in solid cancers, and studies have revealed that low concentrations of nitric oxide (NO) mimetics attenuate hypoxia-induced drug resistance in tumor cells in vitro. Classic NO signaling involves activation of soluble guanylyl cyclase, generation of cyclic GMP (cGMP), and activation of cGMP-dependent protein kinase. Here, we determined whether chemosensitization by NO mimetics requires cGMP-dependent signaling and whether low concentrations of NO mimetics can chemosensitize tumors in vivo. EXPERIMENTAL DESIGN: Survival of human prostate and breast cancer cells was assessed by clonogenic assays following exposure to chemotherapeutic agents. The effect of NO mimetics on tumor chemosensitivity in vivo was determined using a mouse xenograft model of human prostate cancer. Drug efflux in vitro was assessed by measuring intracellular doxorubicin-associated fluorescence. RESULTS: Low concentrations of the NO mimetics glyceryl trinitrate (GTN) and isosorbide dinitrate attenuated hypoxia-induced resistance to doxorubicin and paclitaxel. Similar to hypoxia induced drug resistance, inhibition of various components of the NO signaling pathway increased resistance to doxorubicin, whereas activation of the pathway with 8-bromo-cGMP attenuated hypoxia-induced resistance. Drug efflux was unaffected by hypoxia and inhibitors of drug efflux did not significantly attenuate hypoxia-induced chemoresistance. Compared with mice treated with doxorubicin alone, tumor growth was decreased in mice treated with doxorubicin and a transdermal GTN patch. The presence of GTN and GTN metabolites in plasma samples was confirmed by gas chromatography. CONCLUSION: Tumor hypoxia induces resistance to anticancer drugs by interfering with endogenous NO signaling and reactivation of NO signaling represents a novel approach to enhance chemotherapy. PMID- 17404105 TI - Potent siRNA inhibitors of ribonucleotide reductase subunit RRM2 reduce cell proliferation in vitro and in vivo. AB - PURPOSE: Ribonucleotide reductase (RR) is a therapeutic target for DNA replication-dependent diseases such as cancer. Here, a potent small interfering RNA (siRNA) duplex against the M2 subunit of RR (RRM2) is developed and shown to reduce the growth potential of cancer cells both in vitro and in vivo. EXPERIMENTAL DESIGN: Three anti-RRM2 siRNAs were identified via computational methods, and the potency of these and additional "tiling" duplexes was analyzed in cultured cells via cotransfections using a RRM2-luciferase fusion construct. Knockdown of RRM2 by the best duplex candidates was confirmed directly by Western blotting. The effect of potent duplexes on cell growth was investigated by a real time cell electronic sensing assay. Finally, duplex performance was tested in vivo in luciferase-expressing cells via whole animal bioluminescence imaging. RESULTS: Moderate anti-RRM2 effects are observed from the three duplexes identified by computational methods. However, the tiling experiments yielded an extremely potent duplex (siR2B+5). This duplex achieves significant knockdown of RRM2 protein in cultured cells and has pronounced antiproliferative activity. S.c. tumors of cells that had been transfected with siR2B+5 preinjection grew slower than those of control cells. CONCLUSIONS: An anti-RRM2 siRNA duplex is identified that exhibits significant antiproliferative activity in cancer cells of varying human type and species (mouse, rat, monkey); these findings suggest that this duplex is a promising candidate for therapeutic development. PMID- 17404106 TI - Selective tumor targeting by the hypoxia-activated prodrug AQ4N blocks tumor growth and metastasis in preclinical models of pancreatic cancer. AB - PURPOSE: The antitumor activities and pharmacokinetics of the hypoxia-activated cytotoxin AQ4N and its metabolites were assessed in several preclinical models of pancreatic cancers. EXPERIMENTAL DESIGN: The cytotoxic effects of AQ4N prodrug and its bioreduced form, AQ4, were tested against multiple human tumor cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Nude mice bearing s.c. or orthotopically implanted human BxPC-3 or Panc-1 tumor cells were treated with AQ4N. Tumor growth inhibition, time to progression/end point, and liver metastasis were evaluated in treatment versus control groups. Plasma and tumor levels of AQ4N and its metabolites were quantitated by liquid chromatography-tandem mass spectrometry. RESULTS: In contrast to AQ4N, the bioreduced AQ4 metabolite displayed potent cytotoxicity in many human tumor lines, including those derived from human pancreatic adenocarcinomas. Single agent administration of AQ4N significantly delayed tumor growth, progression, and survival in a manner comparable with gemcitabine in multiple pancreatic tumor models in vivo. Survival increases were accompanied by a reduction in incidence and spread of liver metastasis. Quantitation of AQ4N and its metabolites in tumor bearing mice showed that the prodrug is rapidly cleared from the circulation by 24 h and neither of the bioreduced metabolites was detected in plasma. In contrast, AQ4N readily penetrated BxPC-3 tumors and the cytotoxic AQ4 metabolite rapidly accumulated in tumor tissues at high levels in a dose-dependent fashion. CONCLUSION: AQ4N undergoes rapid and selective conversion into the potent antineoplastic metabolite AQ4 in tumors in vivo and provides proof of principle for the use of hypoxia-activated prodrugs in the treatment against pancreatic cancers. PMID- 17404107 TI - Preclinical studies of TW-37, a new nonpeptidic small-molecule inhibitor of Bcl 2, in diffuse large cell lymphoma xenograft model reveal drug action on both Bcl 2 and Mcl-1. AB - PURPOSE: Overexpression of Bcl-2 protein has been observed in more than 80% of B cell lymphomas, including diffuse large cell lymphoma (DLCL), the most common subtype of non-Hodgkin's lymphoma. We have previously employed the natural product (-)-gossypol to test its therapeutic potential as a small-molecule inhibitor of Bcl-2 for the treatment of B-cell lymphomas. EXPERIMENTAL DESIGN: Recently, we have used a structure-based strategy to design a new class of potent small-molecule inhibitor acting on Bcl-2. One such lead compound is the benzenesulfonyl derivative TW-37, which was designed to target the BH3-binding groove in Bcl-2 where proapoptotic Bcl-2 proteins, such as Bak, Bax, Bid, and Bim bind. RESULTS: In our fluorescence polarization-based binding assays using recombinant Bcl-2, Bcl-X(L), and Mcl-1 proteins, TW-37 binds to Bcl-2, Bcl-X(L), and Mcl-1 with K(i) values of 290, 1,110 and 260 nmol/L, respectively. Hence, TW 37 is a potent inhibitor of Bcl-2 and has >3-fold selectivity over Bcl-X(L). In vitro, TW-37 showed significant antiproliferative effect in a de novo chemoresistant WSU-DLCL(2) lymphoma cell line and primary cells obtained from a lymphoma patient with no effect on normal peripheral blood lymphocytes. Coimmunoprecipitation experiments showed that TW-37 disrupted heterodimer formation between Bax or truncated-Bid and antiapoptotic proteins in the order Mcl-1 > Bcl-2 >> Bcl-X(L). As expected, TW-37 caused apoptotic death. Pre exposure of lymphoma cells to TW-37 significantly enhanced the killing effect of cyclophosphamide-doxorubicin-vincristine-prednisone (CHOP) regimen. The maximum tolerated dose of TW-37 in severe combined immunodeficient (SCID) mice was 40 mg/kg for three i.v. injections when given alone and 20 mg/kg, x3 when given in combination with CHOP. Using WSU-DLCL(2)-SCID mouse xenograft model, the addition of TW-37 to CHOP resulted in more complete tumor inhibition compared with either CHOP or TW-37 alone. CONCLUSIONS: We conclude that the administration of TW-37, as a potent Bcl-2 and Mcl-1 inhibitor, to standard chemotherapy may prove an effective strategy in the treatment of B-cell lymphoma. PMID- 17404108 TI - Menatetrenone, a vitamin K2 analogue, inhibits hepatocellular carcinoma cell growth by suppressing cyclin D1 expression through inhibition of nuclear factor kappaB activation. AB - PURPOSE: Menatetrenone, a vitamin K2 analogue, plays an important role in the production of blood coagulation factors. Menatetrenone has also bee shown to have antineoplastic effects against several cancer cell lines including hepatocellular carcinoma (HCC) cells. However, the mechanisms by which vitamin K2 inhibits HCC cell growth have not bee fully clarified, and we therefore investigated the molecular basis of vitamin K2-induced growth inhibition of HCC cells. EXPERIMENTAL DESIGN: HCC cells were treated with vitamin K2 and the expression of several growth-related genes including cyclin-dependent kinase inhibitors and cyclin D1 was examined at the mRNA and protein levels. A reporter gene assay of the cyclin D1 promoter was done under vitamin K2 treatment. The regulation of nuclear factor kappaB (NF-kappaB) activation was investigated by a NF-kappaB reporter gene assay, an electrophoretic mobility shift assay, a Western blot for phosphorylated IkappaB, and an in vitro kinase assay for IkappaB kinase (IKK). We also examined the effect of vitamin K2 on the growth of HCC cells transfected with p65 or cyclin D1. RESULTS: Vitamin K2 inhibited cyclin D1 mRNA and protein expression in a dose-dependent manner in the HCC cells. Vitamin K2 also suppressed the NF-kappaB binding site-dependent cyclin D1 promoter activity and suppressed the basal, 12-O-tetradecanoylphorbol-13-acetate (TPA)-, TNF-alpha-, and interleukin (IL)-1-induced activation of NF-kappaB binding and transactivation. Concomitant with the suppression of NF-kappaB activation, vitamin K2 also inhibited the phosphorylation and degradation of IkappaBalpha and suppressed IKK kinase activity. Moreover, HCC cells overexpressing cyclin D1 and p65 became resistant to vitamin K2 treatment. CONCLUSION: Vitamin K2 inhibits the growth of HCC cells via suppression of cyclin D1 expression through the IKK/IkappaB/NF-kappaB pathway and might therefore be useful for treatment of HCC. PMID- 17404109 TI - c-Met is a potentially new therapeutic target for treatment of human melanoma. AB - PURPOSE: c-Met is a receptor tyrosine kinase involved in cell growth, invasion, metastases, and angiogenesis. In this study, we investigated the role of c-Met in melanoma biology using a novel small-molecule tyrosine kinase inhibitor SU11274 and small interfering (si) RNA against the receptor. EXPERIMENTAL DESIGN: The effects of SU11274 and c-Met siRNA were studied on proliferation, apoptosis, differentiation, reactive oxygen species, and intracellular signaling. c-Met mutations were examined, and the expression of c-Met and activated c-Met was studied in nevi, primary, and metastatic melanoma. RESULTS: c-Met was expressed in 6:7 melanoma cell lines by immunoblotting. SU11274 inhibited cell growth in all melanoma cell lines by 85% to 98% with an IC(50) between 1 and 2.5 mumol/L and caused apoptosis (12-58%) in five out of six cell lines. siRNA against c-Met inhibited proliferation of melanoma cells by 60%. This is the first study that shows that SU11274 and siRNA induced microphthalmia-associated transcription factor (MITF) and several other melanoma differentiation proteins and a morphologically differentiated phenotype. SU11274 also inhibited reactive oxygen species formation and phosphorylation of c-Met receptor, AKT and S-6 kinase by the hepatocyte growth factor. A new missense c-Met mutation N948S was identified in cell lines and R988C in tumor tissue in the juxtamembrane domain of c-Met. It was found that c-Met was expressed in 88% of melanomas and 15% of nevi, and that c-Met (pY1003) was activated in 21% of human melanomas. CONCLUSION: These results support the role of c-Met in proliferation, apoptosis, differentiation, and tumor progression of melanoma. SU11274 could be used in the therapeutic inhibition of melanoma. PMID- 17404110 TI - Identification of molecular predictors of response in a study of tipifarnib treatment in relapsed and refractory acute myelogenous leukemia. AB - PURPOSE: Microarray technology was used to identify gene expression markers that predict response to the orally available farnesyltransferase inhibitor tipifarnib (Zarnestra, R115777) in acute myelogenous leukemia (AML). EXPERIMENTAL DESIGN: Gene expression profiles from 58 bone marrow samples from a cohort of relapsed and refractory AML patients were analyzed on the Affymetrix U133A gene chip that contains approximately 22,000 genes. RESULTS: Supervised statistical analysis identified eight gene expression markers that could predict patient response to tipifarnib. The most robust gene was the lymphoid blast crisis oncogene (AKAP13), which predicted response with an overall accuracy of 63%. This gene provided a negative predictive value of 93% and a positive predictive value of 31% (increased from 18%). AKAP13 was overexpressed in patients who were resistant to tipifarnib. When overexpressed in the HL60 and THP1 cell lines, AKAP13 increased the resistance to tipifarnib by approximately 5- to 7-fold. CONCLUSION: Diagnostic gene expression signatures may be used to select a group of AML patients that might respond to tipifarnib. PMID- 17404111 TI - Synergism between arsenic trioxide and heat shock protein 90 inhibitors on signal transducer and activator of transcription protein 3 activity--pharmacodynamic drug-drug interaction modeling. AB - PURPOSE: Constitutive signal transducer and activator of transcription 3 (STAT3) activity, observed in approximately 50% of acute myelogenous leukemia cases and associated with adverse treatment outcome, is down-regulated by arsenic trioxide (ATO). Heat shock protein (HSP) 90 is a molecular chaperone involved in signal transduction pathways. We hypothesized that HSP90 inhibitors will potentiate ATO effect on constitutive STAT3 activity and cell killing. One concern was that the effect of ATO and HSP90 inhibitors will result in up-regulation of HSP70, a protein known to inhibit apoptosis. EXPERIMENTAL DESIGN: We have used a semimechanistic pharmacodynamic model to characterize concentration-effect relationships of ATO and HSP90 inhibitors on constitutive STAT3 activity, HSP70 expression, and cell death in a cell line model. RESULTS: Pharmacodynamic interaction of ATO and three HSP90 inhibitors showed synergistic interactions in inhibiting constitutive STAT3 activity and inducing cell death, in spite of a concurrent synergistic up-regulation of HSP70. CONCLUSIONS: These preliminary results provide a basis for studying the combined role of ATO with HSP90 inhibitors in acute myelogenous leukemia with constitutive STAT3 activity. PMID- 17404112 TI - The therapeutic and preventive effect of RRR-alpha-vitamin E succinate on prostate cancer via induction of insulin-like growth factor binding protein-3. AB - PURPOSE: Insulin-like growth factor binding protein-3 (IGFBP-3) is a well-known antiproliferative and proapoptotic molecule in prostate cancer, suggesting that targeting IGFBP-3 might produce clinical benefits. In prostate cancer cells, RRR alpha-vitamin E succinate (VES) inhibits cell proliferation and induces apoptosis, yet the mechanisms remain to be elucidated. We hypothesize that the protective effects of VES in prostate cancer are mediated by IGFBP-3 up regulation. Using prostate cancer models, the involvement of IGFBP-3 in the anticancer effect of VES was investigated. EXPERIMENTAL DESIGN: IGFBP-3 mRNA and protein were determined by real-time PCR and Western blotting in prostate cancer cells, xenografted tumors of nude mice, and prostate tumors of transgenic adenocarcinoma mouse prostate (TRAMP) mice. The serum levels of IGFBP-3 were assessed by ELISA. The importance of IGFBP-3 in VES-mediated antitumor effects was confirmed by small interfering RNA knockdown strategy. RESULTS: We found that VES induced IGFBP-3 mRNA and protein levels in human prostate cancer cell lines. Knockdown of IGFBP-3 by small interfering RNA attenuated VES-induced IGFBP-3 expression and VES-mediated antiproliferative and proapoptotic functions. Furthermore, administration of VES resulted in a significant therapeutic effect on LNCaP and PC3 xenografts and a preventive effect on tumorigenic progression in the TRAMP model without overt toxicity. Notably, the therapeutic and preventive efficacy of VES correlated with increased accumulation of IGFBP-3 in mouse serum as well as in the xenograft tumors and TRAMP prostate samples. Consequently, reduced proliferation and induced apoptosis were witnessed. CONCLUSIONS: VES mediates its therapeutic and preventive effects against prostate cancer at least partially through up-regulating IGFBP-3, which inhibits cell proliferation and promotes cell apoptosis. PMID- 17404113 TI - Identification of a highly effective rapamycin schedule that markedly reduces the size, multiplicity, and phenotypic progression of tobacco carcinogen-induced murine lung tumors. AB - PURPOSE: Human and murine preneoplastic lung lesions induced by tobacco exposure are characterized by increased activation of the Akt/mammalian target of rapamycin (mTOR) pathway, suggesting a role for this pathway in lung cancer development. To test this, we did studies with rapamycin, an inhibitor of mTOR, in A/J mice that had been exposed to the tobacco-specific carcinogen 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK). EXPERIMENTAL DESIGN: Tumorigenesis was induced by i.p. injection of NNK, and rapamycin was administered 1 or 26 weeks after NNK administration. Biomarkers associated with mTOR inhibition were assessed in lung and/or surrogate tissues using immunohistochemistry and immunoblotting. Rapamycin levels were measured using mass spectroscopy. RESULTS: Rapamycin was administered on a daily (5 of 7 days) regimen beginning 26 weeks after NNK decreased tumor size, proliferative rate, and mTOR activity. Multiplicity was not affected. Comparing this regimen with an every-other-day (qod) regimen revealed that rapamycin levels were better maintained with qod administration, reaching a nadir of 16.4 ng/mL, a level relevant in humans. When begun 1 week after NNK, this regimen was well tolerated and decreased tumor multiplicity by 90%. Tumors that did develop showed decreased phenotypic progression and a 74% decrease in size that correlated with decreased proliferation and inhibition of mTOR. CONCLUSIONS: Tobacco carcinogen-induced lung tumors in A/J mice are dependent upon mTOR activity because rapamycin markedly reduced the development and growth of tumors. Combined with the Food and Drug Administration approval of rapamycin and broad clinical experience, these studies provide a rationale to assess rapamycin in trials with smokers at high risk to develop lung cancer. PMID- 17404114 TI - Morin (3,5,7,2',4'-Pentahydroxyflavone) abolishes nuclear factor-kappaB activation induced by various carcinogens and inflammatory stimuli, leading to suppression of nuclear factor-kappaB-regulated gene expression and up-regulation of apoptosis. AB - PURPOSE: Morin is a flavone that exhibits antiproliferative, antitumor, and anti inflammatory effects through a mechanism that is not well understood. Because of the role of transcription factor nuclear factor-kappaB (NF-kappaB) in the control of cell survival, proliferation, tumorigenesis, and inflammation, we postulated that morin mediates its effects by modulating NF-kappaB activation. EXPERIMENTAL DESIGN: We investigated the effect of morin on NF-kappaB pathway activated by inflammatory agents, carcinogens, and tumor promoters. The effect of this flavone on expression of NF-kappaB-regulated gene products involved in cell survival, proliferation, and invasion was also examined. RESULTS: We showed by DNA-binding assay that NF-kappaB activation induced by tumor necrosis factor (TNF), phorbol 12-myristate 13-acetate, lipopolysaccharide, ceramide, interleukin-1, and H(2)O(2) was suppressed by morin; the suppression was not cell type specific. The suppression of NF-kappaB by morin was mediated through inhibition of IkappaBalpha (inhibitory subunit of NF-kappaB) kinase, leading to suppression of phosphorylation and degradation of IkappaBalpha and consequent p65 nuclear translocation. Morin also inhibited the NF-kappaB-dependent reporter gene expression activated by TNF, TNF receptor (TNFR) 1, TNFR1-associated death domain, TNFR-associated factor 2, NF-kappaB-inducing kinase, IkappaB kinase, and the p65 subunit of NF-kappaB. NF-kappaB-regulated gene products involved in cell survival [inhibitor of apoptosis (IAP) 1, IAP2, X chromosome-linked IAP, Bcl-xL, and survivin], proliferation (cyclin D1 and cyclooxygenase-2), and invasion (matrix metalloproteinase-9) were down-regulated by morin. These effects correlated with enhancement of apoptosis induced by TNF and chemotherapeutic agents. CONCLUSION: Overall, our results indicate that morin suppresses the activation of NF-kappaB and NF-kappaB-regulated gene expression, leading to enhancement of apoptosis. This may provide the molecular basis for the ability of morin to act as an anticancer and anti-inflammatory agent. PMID- 17404115 TI - Selective killing of cancer cells by leaf extract of Ashwagandha: identification of a tumor-inhibitory factor and the first molecular insights to its effect. AB - PURPOSE: Ashwagandha is regarded as a wonder shrub of India and is commonly used in Ayurvedic medicine and health tonics that claim its variety of health promoting effects. Surprisingly, these claims are not well supported by adequate studies, and the molecular mechanisms of its action remain largely unexplored to date. We undertook a study to identify and characterize the antitumor activity of the leaf extract of ashwagandha. EXPERIMENTAL DESIGN: Selective tumor-inhibitory activity of the leaf extract (i-Extract) was identified by in vivo tumor formation assays in nude mice and by in vitro growth assays of normal and human transformed cells. To investigate the cellular targets of i-Extract, we adopted a gene silencing approach using a selected small hairpin RNA library and found that p53 is required for the killing activity of i-Extract. RESULTS: By molecular analysis of p53 function in normal and a variety of tumor cells, we found that it is selectively activated in tumor cells, causing either their growth arrest or apoptosis. By fractionation, purification, and structural analysis of the i Extract constituents, we have identified its p53-activating tumor-inhibiting factor as with a none. CONCLUSION: We provide the first molecular evidence that the leaf extract of ashwagandha selectively kills tumor cells and, thus, is a natural source for safe anticancer medicine. PMID- 17404116 TI - The art critic. PMID- 17404117 TI - Magnetic resonance imaging of the thalamus and adhesio interthalamica in twins with schizophrenia. AB - CONTEXT: Abnormalities of the thalamus are thought to be central to the pathophysiology of schizophrenia. These abnormalities include altered structure and shape of the thalamus itself and possibly changes to the adhesio interthalamica (or massa intermedia), the gray matter bridge connecting the 2 thalamic lobes. However, it is not clear to what extent these abnormalities are determined by the genetic liability for schizophrenia. OBJECTIVE: To investigate thalamic volume and the presence of the adhesio interthalamica in monozygotic (MZ) twins concordant or discordant for schizophrenia. DESIGN: Study of MZ twins. SETTING: Patients were drawn from inpatient and outpatient clinics. Twin controls were recruited from a volunteer twin register and through media advertisements. PARTICIPANTS: A total of 123 twins participated: 19 MZ twin pairs concordant for schizophrenia, 15 MZ schizophrenic twins and 16 MZ nonschizophrenic twins drawn from 17 pairs discordant for schizophrenia, and 27 MZ twin pairs without schizophrenia. Groups were matched for age, sex, handedness, level of education, parental socioeconomic status, and ethnicity. MAIN OUTCOME MEASURES: The volume of the thalamus (including right and left hemispheres) was measured (in cubic centimeters) and the presence of the adhesio interthalamica was ascertained from structural magnetic resonance images. RESULTS: Concordant twin pairs displayed significantly reduced thalamic volume compared with control twins, even when covarying for effects of whole-brain volume, age, and sex. There was a significant linear decrease in thalamic volume (control greater than discordant nonschizophrenic greater than discordant schizophrenic greater than concordant). In all groups, right thalamus was larger than left thalamus. There was no difference across groups in the frequency of the adhesio interthalamica. CONCLUSIONS: Volumetric thalamic abnormalities in schizophrenia occur in twin pairs concordant for schizophrenia. These abnormalities may mark the substantial genetic contribution to the illness seen in concordant twin pairs, whereas the adhesio interthalamica is unlikely to be affected in schizophrenia. PMID- 17404118 TI - Association of the brain-derived neurotrophic factor Val66Met polymorphism with reduced hippocampal volumes in major depression. AB - CONTEXT: Brain-derived neurotrophic factor (BDNF) modulates hippocampal plasticity, which is believed to be altered in patients with major depression. OBJECTIVE: To examine the effect of the BDNF Val66Met polymorphism on hippocampal and amygdala volumes in patients with major depression and in healthy control subjects. DESIGN: Cross-sectional comparison between patients and controls. SETTING: Inpatients with major depression from the Department of Psychiatry and Psychotherapy and healthy controls from the community were recruited. PARTICIPANTS: The study population of 120 subjects included 60 patients with major depression and 60 healthy controls. MAIN OUTCOME MEASURES: Using a combined strategy, hippocampal and amygdala volumes were estimated on high-resolution magnetic resonance images, and genotyping was performed for the BDNF Val66Met polymorphism. RESULTS: Patients had significantly smaller hippocampal volumes compared with controls (P = .02). Significantly smaller hippocampal volumes were observed for patients and for controls carrying the Met-BDNF allele compared with subjects homozygous for the Val-BDNF allele (P = .006). With respect to amygdala volumes, no significant differences between patients and controls and no significant main effects for the BDNF Val66Met polymorphism were observed. CONCLUSIONS: These genotype-related alterations suggest that Met-BDNF allele carriers might be at risk to develop smaller hippocampal volumes and may be susceptible to major depression. This study supports findings from animal studies that the hippocampus is involved in brain development and plasticity. PMID- 17404120 TI - Extending the bereavement exclusion for major depression to other losses: evidence from the National Comorbidity Survey. AB - CONTEXT: Symptoms of intense bereavement-related sadness may resemble those of major depressive disorder (MDD) but may not indicate a mental disorder. To avert false-positive diagnoses, DSM criteria for MDD exclude uncomplicated bereavement of brief duration and modest severity. However, the DSM does not similarly exempt depressive reactions to other losses, even when they are uncomplicated in duration and severity. OBJECTIVE: To test the validity of the DSM exclusion of uncomplicated depressive symptoms only in response to bereavement but not in response to other losses. DESIGN: Community-based epidemiological study. PARTICIPANTS: From the National Comorbidity Survey (NCS) of 8098 persons aged 15 to 54 years representative of the US population, we identified individuals who met MDD symptom criteria and whose MDD episodes were triggered by either bereavement (n = 157) or other loss (n = 710). Intervention We divided the bereavement and other loss trigger groups into uncomplicated and complicated cases by applying the NCS algorithm for uncomplicated bereavement to the reactions to other losses. We then compared uncomplicated bereavement and uncomplicated reactions to other losses on a variety of disorder indicators and symptoms. MAIN OUTCOME MEASURES: Nine disorder indicators, as follows: number of symptoms, melancholic depression, suicide attempt, duration of symptoms, interference with life, recurrence, and 3 service use variables. RESULTS: Episodes of uncomplicated depression triggered by bereavement and by other loss have similar symptom profiles and are not significantly different for 8 of 9 disorder indicators. Moreover, uncomplicated reactions, whether triggered by bereavement or other loss, are significantly lower than complicated reactions on almost all disorder indicators. CONCLUSION: The NCS data do not support the validity of uniquely excluding uncomplicated bereavement but not uncomplicated reactions to other losses from MDD diagnosis. PMID- 17404119 TI - Psychosocial treatments for bipolar depression: a 1-year randomized trial from the Systematic Treatment Enhancement Program. AB - CONTEXT: Psychosocial interventions have been shown to enhance pharmacotherapy outcomes in bipolar disorder. OBJECTIVE: To examine the benefits of 4 disorder specific psychotherapies in conjunction with pharmacotherapy on time to recovery and the likelihood of remaining well after an episode of bipolar depression. DESIGN: Randomized controlled trial. SETTING: Fifteen clinics affiliated with the Systematic Treatment Enhancement Program for Bipolar Disorder. Patients A total of 293 referred outpatients with bipolar I or II disorder and depression treated with protocol pharmacotherapy were randomly assigned to intensive psychotherapy (n = 163) or collaborative care (n = 130), a brief psychoeducational intervention. INTERVENTIONS: Intensive psychotherapy was given weekly and biweekly for up to 30 sessions in 9 months according to protocols for family focused therapy, interpersonal and social rhythm therapy, and cognitive behavior therapy. Collaborative care consisted of 3 sessions in 6 weeks. MAIN OUTCOME MEASURES: Outcome assessments were performed by psychiatrists at each pharmacotherapy visit. Primary outcomes included time to recovery and the proportion of patients classified as well during each of 12 study months. RESULTS: All analyses were by intention to treat. Rates of attrition did not differ across the intensive psychotherapy (35.6%) and collaborative care (30.8%) conditions. Patients receiving intensive psychotherapy had significantly higher year-end recovery rates (64.4% vs 51.5%) and shorter times to recovery than patients in collaborative care (hazard ratio, 1.47; 95% confidence interval, 1.08 2.00; P = .01). Patients in intensive psychotherapy were 1.58 times (95% confidence interval, 1.17-2.13) more likely to be clinically well during any study month than those in collaborative care (P = .003). No statistically significant differences were observed in the outcomes of the 3 intensive psychotherapies. CONCLUSIONS: Intensive psychosocial treatment as an adjunct to pharmacotherapy was more beneficial than brief treatment in enhancing stabilization from bipolar depression. Future studies should compare the cost effectiveness of models of psychotherapy for bipolar disorder. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00012558. PMID- 17404121 TI - Second-generation antipsychotic agents in the treatment of acute mania: a systematic review and meta-analysis of randomized controlled trials. AB - CONTEXT: Recommendations of treatment guidelines concerning the use of second generation antipsychotic (SGA) agents for acute mania vary substantially across committees or working groups. Meta-analyses addressing the use of SGAs in the treatment of acute mania are lacking. OBJECTIVE: To conduct a meta-analysis of the efficacy and safety of SGAs in the treatment of acute mania. DATA SOURCES: Randomized controlled trials comparing SGAs with placebo, first-generation antipsychotic drugs, or mood stabilizers (MSs) in the treatment of acute mania were searched for in the PsiTri and MEDLINE databases (last search: May 2006). STUDY SELECTION: The abstracts, titles, and index terms of studies were searched using the following key words: aripiprazole, amisulpride, clozapine, olanzapine, quetiapine, risperidone, ziprasidone, and zotepine in conjunction with mania, manic, and bipolar. DATA EXTRACTION: Data on efficacy, global dropout, dropout due to adverse events, dropout due to inefficacy, weight gain, rate of somnolence, and extrapyramidal symptoms were extracted and combined in a meta analysis. DATA SYNTHESIS: A total of 24 studies with 6187 patients were included. The SGAs were significantly more efficacious than placebo. The analysis demonstrated that adding antipsychotic agents to MS treatment was significantly more effective than treatment with MSs alone. The SGAs displayed efficacy comparable with that of MSs. Some SGAs seemed to induce more extrapyramidal symptoms than placebo. The SGAs were also associated with higher rates of somnolence than placebo. CONCLUSION: Currently available data suggest that combining SGAs and MSs is the most efficacious treatment of acute mania. PMID- 17404122 TI - Parenting and adolescent antisocial behavior and depression: evidence of genotype x parenting environment interaction. AB - BACKGROUND: Little is known about the interplay of genotypes and malleable risk factors in influencing adolescent psychiatric symptoms and disorders. Information on these processes is crucial in designing programs for the prevention of psychiatric disorders. OBJECTIVE: To assess whether latent genetic factors and measured parent-child relationships interact (G x E) in predicting adolescent antisocial behavior and depression. DESIGN: We characterized risk of antisocial behavior and depression in adolescents by means of a genetically informed design. We used in-home questionnaire and observational measures of adolescent outcomes and environmental moderators (parenting), and a latent variable behavior genetic analytic model. SETTING: A nationally distributed sample recruited from random digit dialing and national market panels. PARTICIPANTS: A total of 720 families with at least 2 children, 9 through 18 years old, stratified by genetic relatedness (monozygotic and dizygotic twins, full biological siblings in nondivorced and stepfamilies, and half-siblings and biologically unrelated siblings in stepfamilies). MAIN OUTCOME MEASURES: Antisocial behavior and depressive symptoms. RESULTS: There was an interaction of genotype and both parental negativity and low warmth predicting overall antisocial behavior, as well as aggressive and nonaggressive forms of antisocial behavior, but not depression. Genetic influence was greater for adolescent antisocial behavior when parenting was more negative or less warm. Genotype-environment correlation was partialled out in the analysis and thus did not account for the results. CONCLUSION: This study demonstrates, on the basis of careful measurement and appropriate analytic methods, that a continuous measure of parenting in the normative range moderates the influence of genotype on antisocial behavior. PMID- 17404123 TI - Impact of publicity concerning pediatric suicidality data on physician practice patterns in the United States. AB - CONTEXT: IMS Health Inc data presented by the Food and Drug Administration (FDA) on September 13 and 14, 2004, at a joint meeting of the Center for Drug Evaluation and Research's Psychopharmacologic Drugs Advisory Committee and the FDA's Pediatric Advisory Committee suggested that the number of children and teenagers who were prescribed antidepressants continued to increase in 2004, despite widespread publicity surrounding 2 FDA advisories regarding the potential for pediatric suicidality with selective serotonin reuptake inhibitor use. These results are contradictory to findings from the Medco Health Solutions, Inc, March 2004 analysis of pharmacy benefit claims and a separate subsequent analysis conducted by NDC Health using dispensing data from March 31, 2004, through June 30, 2005. OBJECTIVES: To investigate the contradictory findings and provide additional analyses on the prescribing trends of antidepressants across age groups and physician specialties in the United States. DESIGN: Retail pharmacy prescription data and physician audit data were obtained from Verispan, a joint venture between Quintiles Transnational and McKesson. In addition to examining prescribing trends, a joinpoint regression analysis was conducted to identify the timing for significant changes in prescription use. RESULTS: The analyses suggest that the number of children and teenagers who were prescribed antidepressants has decreased significantly (P = .02) in the wake of widespread publicity surrounding the FDA public health advisories. Another impact of the advisories seems to be a shift in care from "generalists" to psychiatric specialists when it comes to prescribing antidepressants to patients younger than 18 years. Finally, the analyses highlight a slight shift in prescribing toward the non-selective serotonin reuptake inhibitor bupropion hydrochloride, even though it carries the same FDA "black box" warning as the selective serotonin reuptake inhibitors. CONCLUSIONS: The effect on antidepressant prescribing volume observed in our analysis of the Verispan data parallels earlier findings reported by Medco Health Solutions, Inc, and NDC Health that the FDA actions have had a significant effect on the prescribing of antidepressants to children and adolescents. Together, these findings underline the importance of presenting a fair balance within the media due to the significant reach of this channel among prescribing physicians. PMID- 17404124 TI - Prediction of differential adult health burden by conduct problem subtypes in males. AB - CONTEXT: A cardinal feature of the DSM-IV diagnostic criteria for conduct disorder is the distinction between childhood- vs adolescent-onset subtypes. Whether such developmental subtypes exist in the population and have different prognoses should be rigorously tested to inform the DSM-V. OBJECTIVES: To evaluate the epidemiological validity of childhood- vs adolescent-onset conduct problems in a prospective birth cohort, and to assess whether life-course persistent conduct problems are associated with a greater adult health burden. DESIGN, SETTING, AND PARTICIPANTS: Our sample includes 526 male study members in the Dunedin Multidisciplinary Health and Development Study, a 1-year birth cohort (April 1, 1972, through March 30, 1973). Developmental trajectories were defined using prospective ratings of conduct problems at 7, 9, 11, 13, 15, 18, 21, and 26 years of age. MAIN OUTCOME MEASURES: Health burden was assessed as mental and physical health problems at 32 years of age measured via diagnostic interviews and physical examinations. RESULTS: We identified the following 4 developmental subtypes of conduct problems through general growth mixture modeling: (1) childhood-onset/life-course-persistent, (2) adolescent onset, (3) childhood limited, and (4) low. At 32 years of age, study members with the life-course persistent subtype experienced the worst health burden. To a lesser extent, those with the adolescent-onset subtype also experienced health problems. A childhood limited subtype not specified by DSM-IV was revealed; its adult health outcomes were within the range of the cohort norm. CONCLUSIONS: Results support the epidemiological validity of the DSM-IV conduct disorder distinction based on age of onset but highlight the need to also consider long-term persistence to refine diagnosis. Preventing and treating conduct problems has the potential to reduce the adult health burden. PMID- 17404125 TI - Race, ethnicity, and the use of services for mental disorders: results from the National Survey of American Life. AB - CONTEXT: Little is known about differences in the unmet need for mental health service use between African Americans and Caribbean blacks. OBJECTIVE: To extend the National Survey of Black Americans by examining 12-month mental health service use for African Americans and Caribbean blacks from the recently completed National Survey of American Life. DESIGN AND SETTING: National household probability samples of noninstitutionalized African Americans and Caribbean blacks (blacks from Caribbean area countries now living in the United States) conducted between February 2001 and June 2003, using a slightly modified World Mental Health version of the World Health Organization's Composite International Diagnostic Interview. PARTICIPANTS: A total of 3570 African Americans and 1621 Caribbean blacks 18 years and older (N = 5191). MAIN OUTCOME MEASURES: Proportion of respondents with 12-month DSM-IV disorders who sought help in the specialty mental health, general medical, human service, and complementary-alternative medicine treatment sectors. The percentage receiving minimally adequate treatment was also assessed. RESULTS: Overall, 10.1% of respondents used some form of mental heath care services in the past year. Use of services was much higher among those who met criteria for a 12-month DSM-IV disorder (31.9%) than among those who did not (5.4%). Forty-nine percent of respondents with serious mental illness used services, whereas 39.3% had contact with mental health care specialists. The youngest and oldest age groups were least likely to obtain any services. Among African Americans, women were more likely than men to use general medical care and services from any sector. Respondents with the most years of education showed the highest use of services. CONCLUSIONS: The underuse of mental health services among black Americans remains a serious concern. Educational interventions that focus on both consumers and mental health care professionals are needed. PMID- 17404126 TI - Stroke in young adults who abuse amphetamines or cocaine: a population-based study of hospitalized patients. AB - CONTEXT: The abuse of stimulant drugs is increasing in the western United States. Although numerous case reports and animal studies suggest a link with stroke, epidemiologic studies have yielded conflicting results. OBJECTIVE: To test the hypothesis that young adults who abuse amphetamines or cocaine are at a higher risk of stroke. DESIGN, SETTING, AND PARTICIPANTS: Using a cross-sectional design and from a quality indicators' database of 3 148 165 discharges from Texas hospitals, we estimated the secular trends from January 1, 2000, to December 31, 2003, in the abuse of various drugs and of strokes. We developed separate logistic regression models of risk factors for hemorrhagic (n = 937) and ischemic (n = 998) stroke discharges of persons aged 18 to 44 years in 2003, and for mortality risk in patients with stroke. Main Outcome Measure Incidence of stroke using definitions from the Agency for Healthcare Research and Quality's stroke mortality Inpatient Quality Indicator. RESULTS: From 2000 to 2003, the rate of increase was greatest for abuse of amphetamines, followed by cannabis and cocaine. The rate of strokes also increased, particularly among amphetamine abusers. In 812 247 discharges in 2003, amphetamine abuse was associated with hemorrhagic stroke (adjusted odds ratio [OR], 4.95; 95% confidence interval [CI], 3.24-7.55), but not with ischemic stroke; cocaine abuse was associated with hemorrhagic (OR, 2.33; 95% CI, 1.74-3.11) and ischemic (OR, 2.03; 95% CI, 1.48 2.79) stroke. Amphetamine, but not cocaine, abuse was associated with a higher risk of death after hemorrhagic stroke (OR, 2.63; 95% CI, 1.07-6.50). CONCLUSION: Increases in stimulant drug abuse may increase the rate of hospital admissions for strokes and stroke-related mortality. PMID- 17404127 TI - Autism through my eyes. PMID- 17404128 TI - Prenatal and perinatal risk factors for autism: a review and integration of findings. AB - OBJECTIVE: To review the evidence for the presence of prenatal and perinatal factors that affect the risk of autism and autism spectrum disorders. DATA SOURCES: Relevant articles were identified by searching MEDLINE, screening reference lists of original studies, and searching major journals likely to publish epidemiological studies on the topic. STUDY SELECTION: For inclusion in this review, studies required (1) a well-defined sample of cases drawn from population-based registers or cohorts; (2) standardized, prospectively collected obstetric information from birth records or registers; (3) comparison subjects drawn from the general population with information on obstetric complications collected from the same source; and (4) a standardized format for presentation of data, allowing for comparisons among studies. Main Exposures Parental characteristics and obstetric complications. MAIN OUTCOME MEASURES: Rates of autism and autism spectrum disorders. RESULTS: Seven epidemiological studies were identified that fulfilled inclusion criteria. The parental characteristics associated with an increased risk of autism and autism spectrum disorders included advanced maternal age, advanced paternal age, and maternal place of birth outside Europe or North America. The obstetric conditions that emerged as significant fell into 2 categories: (1) birth weight and duration of gestation and (2) intrapartum hypoxia. CONCLUSIONS: Evidence to suggest that parental age and obstetric conditions are associated with an increased risk of autism and autism spectrum disorders is accumulating. Although not proven as independent risk factors for autism, these variables should be examined in future studies that use large, population-based birth cohorts with precise assessments of exposures and potential confounders. PMID- 17404129 TI - Maternal and paternal age and risk of autism spectrum disorders. AB - OBJECTIVE: To explore the association between maternal and paternal age and risk of autism spectrum disorders (ASDs) in offspring. DESIGN: Historical birth cohort study. SETTING: Kaiser Permanente (KP) in Northern California. PARTICIPANTS: All singleton children born at KP from January 1, 1995, to December 31, 1999, were included in the study. We identified 593 children who had ASD diagnoses (International Classification of Diseases, Ninth Revision, Clinical Modification, code 299.0 or 299.8) recorded 2 or more times in KP outpatient databases before May 2005. These children were compared with all 132,251 remaining singleton KP births. Main Exposures Maternal and paternal age at birth of offspring. MAIN OUTCOME MEASURES: Relative risks (RRs) estimated from proportional hazards regression models. Risk of ASDs evaluated in relation to maternal and paternal age, adjusted for each other and for the sex, birth date, and birth order of the child, maternal and paternal educational level, and maternal and paternal race/ethnicity. RESULTS: Risk of ASDs increased significantly with each 10-year increase in maternal age (adjusted RR, 1.31; 95% confidence interval [CI], 1.07 1.62) and paternal age (RR, 1.28; 95% CI, 1.09-1.51). Adjusted RRs for both maternal and paternal age were elevated for children with autistic disorder (maternal age: RR, 1.18; 95% CI, 0.87-1.60; paternal age: RR, 1.34; 95% CI, 1.06 1.69) and children with Asperger disorder or pervasive developmental disorder not otherwise specified (maternal age: RR, 1.45; 95% CI, 1.09-1.93; paternal age: RR, 1.24; 95% CI, 0.99-1.55). Associations with parental age were somewhat stronger for girls than for boys, although sex differences were not statistically significant. CONCLUSION: Advanced maternal and paternal ages are independently associated with ASD risk. PMID- 17404130 TI - The lifetime distribution of the incremental societal costs of autism. AB - OBJECTIVE: To describe the age-specific and lifetime incremental societal costs of autism in the United States. DESIGN: Estimates of use and costs of direct medical and nonmedical care were obtained from a literature review and database analysis. A human capital approach was used to estimate lost productivity. These costs were projected across the life span, and discounted incremental age specific costs were computed. SETTING: United States. PARTICIPANTS: Hypothetical incident autism cohort born in 2000 and diagnosed in 2003. MAIN OUTCOME MEASURES: Discounted per capita incremental societal costs. RESULTS: The lifetime per capita incremental societal cost of autism is $3.2 million. Lost productivity and adult care are the largest components of costs. The distribution of costs over the life span varies by cost category. CONCLUSIONS: Although autism is typically thought of as a disorder of childhood, its costs can be felt well into adulthood. The substantial costs resulting from adult care and lost productivity of both individuals with autism and their parents have important implications for those aging members of the baby boom generation approaching retirement, including large financial burdens affecting not only those families but also potentially society in general. These results may imply that physicians and other care professionals should consider recommending that parents of children with autism seek financial counseling to help plan for the transition into adulthood. PMID- 17404131 TI - Health care expenditures associated with autism spectrum disorders. AB - OBJECTIVE: To examine the health care expenditures associated with autism spectrum disorders (ASDs) in medical care settings. DESIGN: Retrospective analysis of health insurance claims data. SETTING: Administrative claims database for a national sample of privately insured individuals. PARTICIPANTS: Children and adolescents 17 years and younger with a diagnosis of a mental disorder between 2000 and 2004. MAIN OUTCOME MEASURES: Annual inpatient, outpatient, and prescription drug expenditures. RESULTS: Average health care expenditures for individuals with an ASD increased 20.4% from $4965 per patient in 2000 to $5979 per patient in 2004, even after adjustment for inflation. When combined with rising ASD prevalence rates, total expenditures per 10 000 covered lives associated with ASDs increased 142.1% over the 5-year period. Although total expenditures per treated patient were higher for patients with ASDs than for individuals with other mental disorders, ASDs created a smaller burden on health insurers because of their relatively low treated prevalence. CONCLUSIONS: In light of anticipated patterns of earlier identification and more proactive treatment of ASDs in the years to come, the burden of autism on the health care system will continue to increase. Efforts should be made to ensure that access to care for this vulnerable population is not compromised. PMID- 17404132 TI - Risk of autistic disorder in affected offspring of mothers with a glutathione S transferase P1 haplotype. AB - OBJECTIVE: To test whether polymorphisms of the glutathione S-transferase P1 gene (GSTP1) act in the mother during pregnancy to contribute to the phenotype of autistic disorder (AD) in her fetus. DESIGN: Transmission disequilibrium testing (TDT) in case mothers and maternal grandparents. SETTING: Autistic disorder may result from multiple genes and environmental factors acting during pregnancy and afterward. Teratogenic alleles act in mothers during pregnancy to contribute to neurodevelopmental disorders in their offspring; however, only a handful have been identified. GSTP1 is a candidate susceptibility gene for AD because of its tissue distribution and its role in oxidative stress, xenobiotic metabolism, and JNK regulation. PARTICIPANTS: We genotyped GSTP1*G313A and GSTP1*C341T polymorphisms in 137 members of 49 families with AD. All probands received a clinical diagnosis of AD by Autism Diagnostic Interview-Revised and Autism Diagnostic Observation Schedule-Generic testing. MAIN OUTCOME MEASURES: Association of haplotypes with AD was tested by the TDT-Phase program, using the expectation-maximization (EM) algorithm for uncertain haplotypes and for incomplete parental genotypes, with standard measures of statistical significance. RESULTS: The GSTP1*A haplotype was overtransmitted to case mothers (P = .01 [P = .03 using permutation testing]; odds ratio, 2.67 [95% confidence interval, 1.39-5.13]). Results of the combined haplotype and genotype analyses suggest that the GSTP1-313 genotype alone determined the observed haplotype effect. CONCLUSIONS: Overtransmission of the GSTP1*A haplotype to case mothers suggests that action in the mother during pregnancy likely increases the likelihood of AD in her fetus. If this is confirmed and is a result of a gene environment interaction occurring during pregnancy, these findings could lead to the design of strategies for prevention or treatment. PMID- 17404133 TI - Parental compliance after screening social development in toddlers. AB - OBJECTIVES: To examine the prevalence of parents' compliance with follow-up measurements after their child tested positive at a screening to assess problems in social development, as well as to find demographic, screening-related, and child-specific factors associated with parental compliance. DESIGN: Two-stage screening design. SETTING: Utrecht, the Netherlands. PARTICIPANTS: A random population of 31,724 children were screened at well-baby clinics at age 14 to 15 months (screen 1). Three hundred sixty-four children underwent screen 2 (255 children who scored positive at screen 1 [population screening] and 109 children younger than 36 months who were identified by surveillance because of suspected problems in their social development). Main Exposure A 2-stage screening was applied. MAIN OUTCOME MEASURES: Compliance with recommendations of having either a second screening (after screen 1) or clinical evaluation (after screen 2). RESULTS: Of 370 children who tested positive at screen 1, parents of 255 children (69%) complied with screen 2. Three groups were distinguished after screen 2 (n = 173): early compliance (clinical evaluation within 6 months) (68%), late compliance (clinical evaluation after 6 months) (14%), and noncompliance (no clinical evaluation) (18%). Late compliance and noncompliance were more common in parents of younger children and children who were identified via population screening. Parents of children with either relatively high cognitive skills and/or low scores on screening measures were less inclined to comply. CONCLUSIONS: Study results suggest higher effectiveness of surveillance over population screening. Screening may well be applied as a second step after surveillance to identify children who need further clinical evaluation. PMID- 17404134 TI - Heritability of autistic traits in the general population. AB - OBJECTIVE: To explore genetic and environmental influences on individual differences in autistic traits in early adulthood and to test if there is assortative mating (nonrandom partner choice) for autistic traits in the general population. DESIGN: Twin family study using structural equation modeling. SETTING: Population-based twin family sample from the Netherlands. PARTICIPANTS: Twins aged 18 years (n = 370) and their siblings (n = 94); parents of twins (128 couples). Main Outcome Measure Self-reported Autism-Spectrum Quotient (AQ) scores, a quantitative measure of autistic traits. RESULTS: Autistic traits were continuously distributed in the population. Twins and siblings did not significantly differ in AQ scores; men obtained significantly higher AQ scores than women (in twin-sibling sample, P<.001; twin-parent sample, P = .02). Individual differences in endorsement on autistic traits show substantial heritability (57%). No significant shared environmental influences were detected. The genes affecting autistic traits appear to be the same across the sexes. The correlation in AQ score between spouses was low and not significant (Pearson r = .05; P = .59). CONCLUSIONS: Previous general population twin studies reported high heritability for autistic traits in childhood and early adolescence. This study extends these findings to late adolescence and yields no evidence for sex specific genetic influences on autistic traits in later stages of development. As autistic traits show substantial variation in the general population, future genetic studies may be facilitated by measuring autistic traits on a continuous scale like the AQ. No evidence for assortative mating for autistic traits was found, suggesting that, in the general population, there is no passive or active partner selection for autistic traits. PMID- 17404135 TI - A prospective study of response to name in infants at risk for autism. AB - OBJECTIVE: To assess the sensitivity and specificity of decreased response to name at age 12 months as a screen for autism spectrum disorders (ASD) and other developmental delays. DESIGN: Prospective, longitudinal design studying infants at risk for ASD. SETTING: Research laboratory at university medical center. PARTICIPANTS: Infants at risk for autism (55 six-month-olds, 101 twelve-month olds) and a control group at no known risk (43 six-month-olds, 46 twelve-month olds). To date, 46 at-risk infants and 25 control infants have been followed up to 24 months. Intervention Experimental task eliciting response-to-name behavior. MAIN OUTCOME MEASURES: Autism Diagnostic Observation Schedule, Mullen Scales of Early Learning. RESULTS: At age 6 months, there was a nonsignificant trend for control infants to require a fewer number of calls to respond to name than infants at risk for autism. At age 12 months, 100% of infants in the control group "passed," responding on the first or second name call, while 86% in the at risk group did. Three fourths of children who failed the task were identified with developmental problems at age 24 months. Specificity of failing to respond to name was 0.89 for ASD and 0.94 for any developmental delay. Sensitivity was 0.50 for ASD and 0.39 for any developmental delay. CONCLUSIONS: Failure to respond to name by age 12 months is highly suggestive of developmental abnormality but does not identify all children at risk for developmental problems. Lack of responding to name is not universal among infants later diagnosed with ASD and/or other developmental delays. Poor response to name may be a trait of the broader autism phenotype in infancy. PMID- 17404136 TI - Early social-communicative and cognitive development of younger siblings of children with autism spectrum disorders. AB - OBJECTIVE: To compare the early social-communicative development of younger siblings of children with autism spectrum disorders (ASDs) with that of younger siblings of children with typical development, using parental report and child based measures. DESIGN: Group comparison. SETTING: Vanderbilt University, between July 1, 2003, and July 31, 2006. PARTICIPANTS: Younger siblings of children with ASD (n = 64) and younger siblings of children with typical development (n = 42) between the ages of 12 and 23 months (mean, 16 months). Main Exposure Having a sibling with an ASD. OUTCOME MEASURES: Child-based measures included a cognitive assessment; an interactive screening tool assessing play, imitation, and communication; and a rating of autism symptoms. Parental report measures were an interview of social-communicative interactions and a questionnaire assessing language and communication skills. RESULTS: Younger siblings of children with ASD demonstrated weaker performance in nonverbal problem solving (mean difference [MD], 5.91; 95% confidence interval [CI], 2.48-9.34), directing attention (MD, 0.52; 95% CI, 0.07-0.97), understanding words (MD, 33.30; 95% CI, 3.11-63.48), understanding phrases (MD, 4.56; 95% CI, 1.85-7.27), gesture use (MD, 1.49; 95% CI, 0.51-2.47), and social-communicative interactions with parents (MD, 1.32; 95% CI, 0.27-2.37), and had increased autism symptoms (MD, 2.54; 95% CI, 1.05-4.03), relative to control siblings. A substantial minority of the ASD sibling group exhibited lower performance relative to controls. Significant correlations between child-based measures and parental reports assessing similar constructs were found (r = -0.74 to 0.53; P range, .000-.002). CONCLUSION: The weaker performance found for children in the ASD sibling group may represent early emerging features of the broader autism phenotype, thus highlighting the importance of developmental surveillance for younger siblings. PMID- 17404137 TI - Understanding autism: parents and pediatricians in historical perspective. AB - Both primary care providers and subspecialists in pediatrics encounter families who are actively involved in the diagnosis and treatment of their children. Parents of children with an autism spectrum disorder in particular are often aware of scientific issues, and their expertise and desire for a medical cure for autism sometimes put them at odds with the medical team. We investigated the role of parents and advocates in autism research and treatment over the last 50 years. Our review of scientific publications and archival sources documents how parents and advocacy groups have done the following: (1) organized research funding; (2) constructed clinical research networks; (3) suggested new avenues for research; (4) popularized empirically based therapies; and (5) anticipated paradigmatic shifts in the understanding of autism. We believe that this historical account will help pediatricians and researchers recognize that families can contribute to expert understanding of complex medical conditions such as autism and that the existence of partnerships with families of children with autism is a critical component of future research and treatment programs. PMID- 17404138 TI - Comparison of indicators for a primary care medical home between children with autism or asthma and other special health care needs: National Survey of Children's Health. AB - OBJECTIVE: To assess the extent to which parents of children with autism compared with parents of children with asthma or other special health care needs report receiving primary care for their child consistent with the American Academy of Pediatrics medical home model. DESIGN: Population-based cross-sectional study. SETTING: National Survey for Children's Health 2003-2004 telephone interview. PARTICIPANTS: Parents of 495 children with autism, parents of 6716 children with asthma, and parents of 11,403 children with other special health care needs without asthma. Main Exposures Autism and other special health care needs including asthma. MAIN OUTCOME MEASURES: Medical home score and components of care, as follows: personal provider and preventive; family-centered, compassionate, and culturally appropriate; accessible; comprehensive; and coordinated. RESULTS: The odds of parents reporting care consistent with that in a medical home were less likely for children with autism (odds ratio, 0.45; 95% confidence interval, 0.30-0.66) and more likely for children with asthma (odds ratio, 1.17; 95% confidence interval, 1.06-1.30) compared with children with other special health care needs (1 [reference]). These differences persisted even after controlling for condition severity, personal characteristics, and insurance status. Specific components of a medical home less prevalent among children with autism than among children with other special health care needs included family centered, comprehensive, and coordinated care. CONCLUSION: Although we could not evaluate the reasons why, a large percentage of children with autism do not receive primary care consistent with that in a medical home. PMID- 17404139 TI - Revisiting parental concerns in the age of autism spectrum disorders: the need to help parents in the face of uncertainty. PMID- 17404140 TI - Despite major challenges, autism research continues to offer hope. PMID- 17404142 TI - Addressing the crisis in access to autism treatment using health care improvement science. PMID- 17404143 TI - Finding effective screening instruments for autism using bayes theorem. PMID- 17404144 TI - The role of pediatricians who care for children with autism. PMID- 17404145 TI - Closing facilities for children: an unrealistic position. PMID- 17404146 TI - Growth attenuation by commission and omission may be ethically justifiable in children with profound disabilities. PMID- 17404147 TI - Disabling children with disabilities. PMID- 17404148 TI - Control of early events in olfactory processing by adult neurogenesis. AB - The mature brain needs to have flexible control over behavior in the face of ever changing needs. It achieves this control through morphological and physiological changes at the level of molecules, spines, dendrites, and axons and through processes of adult neurogenesis, entire cells. The functional maturation of newly generated cells in the adult forebrain involves the expression of neurotransmitter receptors before synaptic activity and excitatory gamma aminobutyric acid (GABAergic) influences prior to glutamatergic input. The production of new cells for incorporation into neural circuits that are already up and running gives rise to a unique situation that may require epigenetic regulation. However, once mature, new neurons must carve out a niche among more established cells to be useful. How do they survive and what are they used for? Recent studies have revealed that adult neurogenesis alters the olfactory bulb at all levels, from single cells to the network and system levels. It has also been suggested that cell turnover may be particularly beneficial for the processing of new information in dynamic networks. However, elucidating the functional meaning of adult neurogenesis must wait for the development of new paradigms to eliminate the pool of newly generated neurons but sparing the preexisting ones. Nevertheless, there is already considerable correlative evidence to indicate that adult neurogenesis is a plastic mechanism by which the performance of the brain can be optimized in a given environment. PMID- 17404149 TI - Why have neurogenesis in adult olfactory systems? The Presidential Symposium at the 2006 AChemS Conference. PMID- 17404151 TI - The olfactory pathway of decapod crustaceans--an invertebrate model for life-long neurogenesis. AB - The first part of this review includes a short description of the cellular and morphological organization of the olfactory pathway of decapod crustaceans, followed by an overview of adult neurogenesis in this pathway focusing on the olfactory lobe (OL), the first synaptic relay in the brain. Adult neurogenesis in the central olfactory pathway has the following characteristics. 1) It is present in all the diverse species of decapod crustaceans so far studied. 2) In all these species, projection neurons (PNs), which have multiglomerular dendritic arborizations, are generated. 3) Neurons are generated by one round of symmetrical cell divisions of a small population of immediate precursor cells that are located in small proliferation zones at the inner margin of the respective soma clusters. 4) The immediate precursor cells in each soma cluster appear to be generated by repeated cell divisions of one or few neuronal stem cells that are located outside of the proliferation zone. 5) These neuronal stem cells are enclosed in a highly structured clump of small glial-like cells, which likely establishes a specific microenvironment and thus can be regarded as a stem cell niche. 6) Diverse internal and external factors, such as presence of olfactory afferents, age, season of the year, and living under constant and deprived conditions modulate the generation and/or survival of new neurons. In the second part of this review, I address the question why in decapod crustaceans adult neurogenesis persists in the visual and olfactory pathways of the brain but is lacking in all other mechanosensory-chemosensory pathways. Due to the indeterminate growth of most adult decapod crustaceans, new sensory neurons of all modalities (olfaction and chemo-, mechano-, and photoreception) are continuously added during adulthood and provide an ever-increasing sensory input to all primary sensory neuropils of the central nervous system. From these facts, I conclude that adult neurogenesis in the brain cannot simply be a mechanism to accommodate increasing sensory input and propose instead that it is causally linked to the specific "topographic logic" of information processing implemented in the sensory neuropils serving different modalities. For the presumptive odotopic type of information processing in the OL, new multiglomerular PNs allow interconnection of novel combinations of spatially unrelated input channels (glomeruli), whose simultaneous activation by specific odorants is the basis of odor coding. Thus, adult neurogenesis could provide a unique way to increase the resolution of odorant quality coding and allow adaptation of the olfactory system of these long-lived animals to ever-changing odor environments. PMID- 17404150 TI - Understanding the regulation and function of adult neurogenesis: contribution from an insect model, the house cricket. AB - Since the discovery of adult neurogenesis, a major issue is the role of newborn neurons and the function-dependent regulation of adult neurogenesis. We decided to use an animal model with a relatively simple brain to address these questions. In the adult cricket brain as in mammals, new neurons are produced throughout life. This neurogenesis occurs in the main integrative centers of the insect brain, the mushroom bodies (MBs), where the neuroblasts responsible for their formation persist after the imaginal molt. The rate of production of new neurons is controlled not only by internal cues such as morphogenetic hormones but also by external environmental cues. Adult crickets reared in an enriched sensory environment experienced an increase in neuroblast proliferation as compared with crickets reared in an impoverished environment. In addition, unilateral sensory deprivation led to reduced neurogenesis in the MB ipsilateral to the lesion. In search of a functional role for the new cells, we specifically ablated MB neuroblasts in young adults using brain-focused gamma ray irradiation. We developed a learning paradigm adapted to the cricket, which we call the "escape paradigm." Using this operant associative learning test, we showed that crickets lacking neurogenesis exhibited delayed learning and reduced memory retention of the task when olfactory cues were used. Our results suggest that environmental cues are able to influence adult neurogenesis and that, in turn, newly generated neurons participate in olfactory integration, optimizing learning abilities of the animal, and thus its adaptation to its environment. Nevertheless, odor learning in adult insects cannot always be attributed to newly born neurons because neurogenesis is completed earlier in development in many insect species. In addition, many of the irradiated crickets performed significantly better than chance on the operant learning task. PMID- 17404152 TI - A phase III randomised trial comparing sequential chemotherapy using cisplatin based regimen and paclitaxel to cisplatin-based chemotherapy alone in advanced non-small-cell lung cancer. AB - BACKGROUND: The purpose of this study is to determine whether in advanced non small-cell lung cancer (NSCLC), the sequential administration of cisplatin-based chemotherapy and paclitaxel (Taxol) is superior to a cisplatin-based chemotherapy, followed by paclitaxel as salvage treatment. PATIENTS AND METHODS: A total of 485 chemotherapy naive patients with advanced NSCLC were treated with three courses of GIP (gemcitibine + ifosfamide + cisplatin), consisting of cisplatin 50 mg/m(2) on day 1, ifosfamide 3 g/m(2) on day 1 and gemcitabine 1 g/m(2) on days 1 and 8. Patients with nonprogressive disease were then randomised to further similar courses of GIP or courses of paclitaxel (225 mg/m(2) over 3 h every 3 weeks). RESULTS: Objective response or nonprogression after induction GIP occurred in 174 and 115 patients, respectively. After randomisation, there were 140 patients in the GIP arm and 141 in the paclitaxel arm. In terms of postrandomisation survival, there was no statistically significant difference (P = 0.17) between the two arms. Median times were 9.7 [95% confidence interval (CI) 7.8-11.6] and 11.9 (95% CI 9.4-14.3) months for paclitaxel and GIP, respectively. Multivariate analysis demonstrated that sex and haemoglobin were independent prognostic factors. After adjustment for these factors, the observed hazard ratio was 0.81 (95% CI 0.63-1.04) in favour of GIP (P = 0.10). Toxicity was tolerable; there was a significantly higher rate of grades III/IV thrombocytopenia with GIP and more alopecia with paclitaxel. CONCLUSION: Sequential chemotherapy using cisplatin-based regimen followed by paclitaxel does not result in better outcome than cisplatin-based chemotherapy using taxane as salvage treatment. PMID- 17404153 TI - Cathepsin L deficiency reduces diet-induced atherosclerosis in low-density lipoprotein receptor-knockout mice. AB - BACKGROUND: Remodeling of the arterial extracellular matrix participates importantly in atherogenesis and plaque complication. Increased expression of the elastinolytic and collagenolytic enzyme cathepsin L (Cat L) in human atherosclerotic lesions suggests its participation in these processes, a hypothesis tested here in mice. METHODS AND RESULTS: We generated Cat L and low density lipoprotein receptor (LDLr) double-deficient (LDLr-/- Cat L-/-) mice by crossbreeding Cat L-null (Cat L-/-) and LDLr-deficient (LDLr-/-) mice. After 12 and 26 weeks of a Western diet, LDLr-/- Cat L-/- mice had significantly smaller atherosclerotic lesions and lipid cores compared with littermate control LDLr-/- Cat L+/- and LDLr-/- Cat L+/+ mice. In addition, lesions from the compound mutant mice showed significantly reduced levels of collagen, medial elastin degradation, CD4+ T cells, macrophages, and smooth muscle cells. Mechanistic studies showed that Cat L contributes to the degradation of extracellular matrix elastin and collagen by aortic smooth muscle cells. Smooth muscle cells from LDLr-/- Cat L-/- mice or those treated with a Cat L-selective inhibitor demonstrated significantly less degradation of elastin and collagen and delayed transmigration through elastin in vitro. Cat L deficiency also significantly impaired monocyte and T lymphocyte transmigration through a collagen matrix in vitro, suggesting that blood-borne leukocyte penetration through the arterial basement membrane requires Cat L. Cysteine protease active site labeling demonstrated that Cat L deficiency did not affect the activity of other atherosclerosis-associated cathepsins in aortic smooth muscle cells and monocytes. CONCLUSIONS: Cat L directly participates in atherosclerosis by degrading elastin and collagen and regulates blood-borne leukocyte transmigration and lesion progression. PMID- 17404154 TI - Paraoxonase-2 reduces oxidative stress in vascular cells and decreases endoplasmic reticulum stress-induced caspase activation. AB - BACKGROUND: In the vascular system, elevated levels of reactive oxygen species (ROS) produce oxidative stress and predispose to the development of atherosclerosis. Therefore, it is important to understand the systems producing and those scavenging vascular ROS. Here, we analyzed the ROS-reducing capability of paraoxonase-2 (PON2) in different vascular cells and its involvement in the endoplasmic reticulum stress pathway known as the unfolded protein response. METHODS AND RESULTS: Quantitative real-time polymerase chain reaction and Western blotting revealed that PON2 is equally expressed in vascular cells and appears in 2 distinct glycosylated isoforms. By determining intracellular ROS, we show that overexpression of PON2 markedly reduced ROS, whereas its knockdown increased ROS levels significantly. Using microscopic and biochemical methods, we found PON2 mainly in the nuclear membrane and endoplasmic reticulum. Furthermore, PON2 expression was induced at both the promoter and protein levels by endoplasmic reticulum stress pathway unfolded protein response. This pathway may promote both apoptotic and survival mechanisms. Functionally, PON2 reduced unfolded protein response-accompanying oxidative stress and unfolded protein response-derived caspase activation. CONCLUSIONS: We suggest that PON2 represents an endogenous defense mechanism against vascular oxidative stress and unfolded protein response induced cell death, thereby contributing to the prevention of atherosclerosis. PMID- 17404155 TI - Recruitment of compensatory pathways to sustain oxidative flux with reduced carnitine palmitoyltransferase I activity characterizes inefficiency in energy metabolism in hypertrophied hearts. AB - BACKGROUND: Transport rates of long-chain free fatty acids into mitochondria via carnitine palmitoyltransferase I relative to overall oxidative rates in hypertrophied hearts remain poorly understood. Furthermore, the extent of glucose oxidation, despite increased glycolysis in hypertrophy, remains controversial. The present study explores potential compensatory mechanisms to sustain tricarboxylic acid cycle flux that resolve the apparent discrepancy of reduced fatty acid oxidation without increased glucose oxidation through pyruvate dehydrogenase complex in the energy-poor, hypertrophied heart. METHODS AND RESULTS: We studied flux through the oxidative metabolism of intact adult rat hearts subjected to 10 weeks of pressure overload (hypertrophied; n=9) or sham operation (sham; n=8) using dynamic 13C-nuclear magnetic resonance. Isolated hearts were perfused with [2,4,6,8,10,12,14,16-(13)C8] palmitate (0.4 mmol/L) plus glucose (5 mmol/L) in a 14.1-T nuclear magnetic resonance magnet. At similar tricarboxylic acid cycle rates, flux through carnitine palmitoyltransferase I was 23% lower in hypertrophied (P<0.04) compared with sham hearts and corresponded to a shift toward increased expression of the L-carnitine palmitoyltransferase I isoform. Glucose oxidation via pyruvate dehydrogenase complex did not compensate for reduced palmitate oxidation rates. However, hypertrophied rats displayed an 83% increase in anaplerotic flux into the tricarboxylic acid cycle (P<0.03) that was supported by glycolytic pyruvate, coincident with increased mRNA transcript levels for malic enzyme. CONCLUSIONS: In cardiac hypertrophy, fatty acid oxidation rates are reduced, whereas compensatory increases in anaplerosis maintain tricarboxylic acid cycle flux and account for a greater portion of glucose oxidation than previously recognized. The shift away from acetyl coenzyme A production toward carbon influx via anaplerosis bypasses energy, yielding reactions contributing to a less energy-efficient heart. PMID- 17404156 TI - Granulocyte colony-stimulating factor and granulocyte macrophage colony stimulating factor exacerbate atherosclerosis in apolipoprotein E-deficient mice. AB - BACKGROUND: Recent studies have suggested a potential contribution of bone marrow derived progenitor cells to vascular repair. Preliminary clinical studies have explored the possibility that mobilization of progenitor cells with granulocyte macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) can affect vascular repair. However, it is not known whether the short-term administration of G-CSF or GM-CSF exerts beneficial effects on atherosclerosis. METHODS AND RESULTS: Apolipoprotein E-deficient mice were treated with either GM-CSF or G-CSF at a dose of 10 microg x kg(-1) x d(-1) s.c. administered daily for 5 days per week on alternating weeks for a total of 20 doses over an 8-week treatment period. We found that in animals maintained on a high-fat diet, both G-CSF and GM-CSF actually demonstrated an increase in atherosclerotic lesion extent. The increase in atherosclerotic extent was not associated with an increase in either inflammatory cells or expression of proinflammatory genes. Interestingly, adventitial vascularity significantly increased, suggesting a mechanistic role for vasa vasorum neovascularization. CONCLUSIONS: These findings demonstrate that in this animal model of atherosclerosis, not only did administration of G-CSF or GM-CSF fail to demonstrate any beneficial therapeutic effect, but both resulted in a worsening of atherosclerosis. PMID- 17404157 TI - Coronary collaterals remain recruitable after percutaneous intervention. AB - BACKGROUND: Rapid loss of collateral support has been reported after percutaneous coronary intervention (PCI), leaving the myocardium susceptible to subsequent infarction. However, well-developed collaterals have been found in normal hearts, suggesting that collaterals exist even in the absence of an ischemic stimulus. We assessed the plasticity and determinants of collateral supply after PCI. METHODS AND RESULTS: Collateral flow index (CFI) was calculated in 60 patients as (P(w) P(v))/(P(a)-P(v)) by measurement of aortic (P(a)), central venous (P(v)), and coronary wedge (P(w)) pressures. CFI was reassessed during transient balloon occlusion 5 minutes and 24 hours after PCI in the first 29 patients and at 6 months in the subsequent 25 patients. We also evaluated the relationship between collateral supply, lesion characteristics, and circulating hemopoietic cells numbers before and after successful PCI. CFI at baseline was 0.23+/-0.10, with no change 5 minutes and 1 day later (0.21+/-0.12, P=0.62; and 0.22+/-0.11, P=0.96, respectively). At 6 months, CFI was 0.14+/-0.07 or 63+/-27% of the baseline value (P<0.001). CFI was proportional to severity of the coronary lesion at baseline (r=0.63, P<0.0001) but not 6 months after PCI (r=-0.04, P=0.87). The number of circulating CD133+ and CD34+ cells was associated with CFI 6 months after PCI (CD133, r=0.59, P=0.035; CD34, r=0.63, P=0.037). CONCLUSIONS: Coronary collateral flow remains undiminished for at least 24 hours after successful PCI. Functional collateral support subsequently declines but does not regress completely. PMID- 17404158 TI - Clinical aspects and prognosis of Brugada syndrome in children. AB - BACKGROUND: Brugada syndrome is an arrhythmogenic disease characterized by an ECG pattern of ST-segment elevation in the right precordial leads and augmented risk of sudden cardiac death. Little is known about the clinical presentation and prognosis of this disease in children. METHODS AND RESULTS: Thirty children affected by Brugada syndrome who were <16 years of age (mean, 8+/-4 years) were included. All patients displayed a type I ECG pattern before or after drug provocation challenge. Diagnosis of Brugada syndrome was made under the following circumstances: aborted sudden death (n=1), syncope of unexplained origin (n=10), symptomatic supraventricular tachycardia (n=1), suspicious ECG (n=1), and family screening for Brugada syndrome (n=17). Syncope was precipitated by fever in 5 cases. Ten of 11 symptomatic patients displayed a spontaneous type I ECG. An implantable cardioverter-defibrillator was implanted in 5 children; 4 children were treated with hydroquinidine; and 1 child received a pacemaker because of symptomatic sick sinus syndrome. During a mean follow-up of 37+/-23 months, 1 child experienced sudden cardiac death, and 2 children received an appropriate implantable cardioverter-defibrillator shock; all of them were symptomatic and had manifested a type I ECG spontaneously. One child had a cardioverter defibrillator infection that required explantation of the defibrillator. CONCLUSIONS: In the largest population of children affected by Brugada syndrome described to date, fever represented the most important precipitating factor for arrhythmic events, and as in the adult population, the risk of arrhythmic events was higher in previously symptomatic patients and in those displaying a spontaneous type I ECG. PMID- 17404159 TI - Cardiac structure and ventricular-vascular function in persons with heart failure and preserved ejection fraction from Olmsted County, Minnesota. AB - BACKGROUND: Mechanisms purported to contribute to the pathophysiology of heart failure with normal ejection fraction (HFnlEF) include diastolic dysfunction, vascular and left ventricular systolic stiffening, and volume expansion. We characterized left ventricular volume, effective arterial elastance, left ventricular end-systolic elastance, and left ventricular diastolic elastance and relaxation noninvasively in consecutive HFnlEF patients and appropriate controls in the community. METHODS AND RESULTS: Olmsted County (Minn) residents without cardiovascular disease (n=617), with hypertension but no heart failure (n=719), or with HFnlEF (n=244) were prospectively enrolled. End-diastolic volume index was determined by echo Doppler. End-systolic elastance was determined using blood pressure, stroke volume, ejection fraction, timing intervals, and estimated normalized ventricular elastance at end diastole. Tissue Doppler e' velocity was used to estimate the time constant of relaxation. End-diastolic volume (EDV) and Doppler-derived end-diastolic pressure (EDP) were used to derive the diastolic curve fitting (alpha) and stiffness (beta) constants (EDP=alphaEDVbeta). Comparisons were adjusted for age, sex, and body size. HFnlEF patients had more severe renal dysfunction, yet smaller end-diastolic volume index and cardiac output and increased EDP compared with both hypertensive and healthy controls. Arterial elastance and ventricular end-systolic elastance were similarly increased in hypertensive controls and HFnlEF patients compared with healthy controls. In contrast, HFnlEF patients had more impaired relaxation and increased diastolic stiffness compared with either control group. CONCLUSIONS: From these cross-sectional observations, we speculate that the progression of diastolic dysfunction plays a key role in the development of heart failure symptoms in persons with hypertensive heart disease. PMID- 17404160 TI - Protective role of endogenous erythropoietin system in nonhematopoietic cells against pressure overload-induced left ventricular dysfunction in mice. AB - BACKGROUND: Erythropoietin (Epo) receptors (EpoRs) are expressed in the heart. We have recently demonstrated that the endogenous Epo-EpoR system plays an important protective role in myocardial ischemia in mice and humans. In the present study, we tested our hypothesis that the endogenous Epo-EpoR system in nonhematopoietic cells also plays a protective role against pressure overload-induced cardiac dysfunction in vivo. METHODS AND RESULTS: Transgene-rescued EpoR-null mutant mice (EpoR-/-(rescued)) that express EpoR exclusively in the hematopoietic cells were subjected to transverse aortic constriction (TAC). At 1 week after TAC, left ventricular weight and lung weight were significantly increased in EpoR-/ (rescued) mice compared with wild-type mice, although the fibrotic area was comparably increased after TAC in the 2 genotypes. In the EpoR-/-(rescued) mice with TAC, left ventricular end-diastolic diameter was significantly increased, left ventricular fractional shortening was significantly decreased, and survival rate was significantly decreased compared with wild-type mice with TAC. Phosphorylation of STAT3 at 5 hours and 1 week after TAC and that of p38 at 5 hours after TAC were significantly increased in wild-type mice but not in EpoR-/ (rescued) mice. Vascular endothelial growth factor protein expression and capillary density in left ventricular myocardium were significantly decreased in EpoR-/-(rescued) mice with TAC compared with wild-type mice with TAC. CONCLUSIONS: These results suggest that the endogenous Epo-EpoR system in the nonhematopoietic cells plays an important protective role against pressure overload-induced cardiac dysfunction in vivo. PMID- 17404161 TI - Mutation of the circadian clock gene Per2 alters vascular endothelial function. AB - BACKGROUND: The circadian clock regulates biological processes including cardiovascular function and metabolism. In the present study, we investigated the role of the circadian clock gene Period2 (Per2) in endothelial function in a mouse model. METHODS AND RESULTS: Compared with the wild-type littermates, mice with Per2 mutation exhibited impaired endothelium-dependent relaxations to acetylcholine in aortic rings suspended in organ chambers. During transition from the inactive to active phase, this response was further increased in the wild type mice but further decreased in the Per2 mutants. The endothelial dysfunction in the Per2 mutants was also observed with ionomycin, which was improved by the cyclooxygenase inhibitor indomethacin. No changes in the expression of endothelial acetylcholine-M3 receptor or endothelial nitric oxide synthase protein but increased cyclooxygenase-1 (not cyclooxygenase-2) protein levels were observed in the aortas of the Per2 mutants. Compared with Per2 mutants, a greater endothelium-dependent relaxation to ATP was observed in the wild-type mice, which was reduced by indomethacin. In quiescent aortic rings, ATP caused greater endothelium-dependent contractions in the Per2 mutants than in the wild-type mice, contractions that were abolished by indomethacin. The endothelial dysfunction in the Per2 mutant mice is not associated with hypertension or dyslipidemia. CONCLUSIONS: Mutation in the Per2 gene in mice is associated with aortic endothelial dysfunction involving decreased production of NO and vasodilatory prostaglandin(s) and increased release of cyclooxygenase-1-derived vasoconstrictor(s). The results suggest an important role of the Per2 gene in maintenance of normal cardiovascular functions. PMID- 17404162 TI - Relative value of inflammatory, hemostatic, and rheological factors for incident myocardial infarction and stroke: the Edinburgh Artery Study. AB - BACKGROUND: The aim of our present study was to compare the association of a wide range of 17 biomarkers of inflammation, hemostasis, and blood rheology with incident heart disease and stroke after accounting for an indicator of subclinical atherosclerotic disease and traditional risk factors and also to determine their incremental predictive ability. METHODS AND RESULTS: We used data from the Edinburgh Artery Study, a population cohort study started in 1987 that comprised 1592 men and women aged 55 to 74 years. Subjects were followed for a mean of 17 years, and 416 of them suffered at least 1 cardiovascular event. In analyses adjusted for cardiovascular risk factors and history of cardiovascular disease (CVD): C-reactive protein, interleukin-6, fibrinogen, fibrin D-dimer, tissue plasminogen activator (t-PA), leukocyte elastase, and lipoprotein(a) (all P<0.01), as well as von Willebrand factor and plasma viscosity (both P<0.05), had significant hazard ratios for incident CVD. Further adjustment for a measure of subclinical atherosclerosis (ankle brachial index) had little impact on these associations. The hazard ratios (95% CI) for incident CVD between top and bottom tertiles in the latter analysis were 1.78 (1.30 to 2.45) for C-reactive protein, 1.85 (1.33 to 2.58) for interleukin-6, and 1.76 (1.35 to 2.31) for fibrinogen. Single biomarkers provided little additional discrimination of incident CVD to that obtained from cardiovascular risk factors and the ankle brachial index. An incremental score of multiple markers [interleukin-6, t-PA, intercellular adhesion molecule 1, and lipoprotein(a)] provided some added discrimination. CONCLUSIONS: Several "novel" risk factors predicted CVD after adjustments for conventional risk factors and also for a measure of asymptomatic disease. However, their incremental predictive ability was modest and their clinical utility remains uncertain. PMID- 17404163 TI - Optimal treatment of obesity-related hypertension: the Hypertension-Obesity Sibutramine (HOS) study. AB - BACKGROUND: Current guidelines for the treatment of hypertension do not provide specific recommendations for obese hypertensive patients. To identify an optimal treatment regimen for obese hypertensive patients, we studied the interactions between a drug-based weight loss approach by sibutramine and different antihypertensive drug regimens. METHODS AND RESULTS: This was a prospective, 16 week double-blind placebo-controlled randomized multicenter study in 171 obese hypertensive patients. After a 2-week run-in period, patients receiving 1 of the 3 antihypertensive combination therapies (felodipine 5 mg/ramipril 5 mg [n=57], verapamil 180 mg/trandolapril 2 mg [n=55], or metoprolol succinate 95 mg/hydrochlorothiazide 12.5 mg [metoprolol/hydrochlorothiazide; n=59]) were assigned randomly to sibutramine (15 mg) or placebo. Sibutramine treatment resulted in a significantly greater decrease in body weight, body mass index, and waist circumference and a significant increase in diastolic blood pressure during 24-hour blood pressure monitoring compared with placebo treatment. Sibutramine induced weight loss and reduction of visceral obesity were markedly attenuated in the metoprolol/hydrochlorothiazide group compared with the other groups. Consistently, improvement in glucose tolerance and hypertriglyceridemia by sibutramine was abrogated in the cohort treated with metoprolol/hydrochlorothiazide compared with the other groups. CONCLUSIONS: The present study demonstrates for the first time that an antihypertensive combination therapy regimen with angiotensin-converting enzyme inhibitors and calcium channel blockers is more advantageous than a beta-blocker/diuretic-based regimen in supporting the weight-reducing actions and concomitant metabolic changes induced by sibutramine in obese hypertensive patients. These data may help to develop future comprehensive treatment strategies and guidelines for this high-risk patient population. PMID- 17404164 TI - Images in cardiovascular medicine. A ring in the heart: calcified left ventricular aneurysm. PMID- 17404165 TI - Images in cardiovascular medicine. Parvovirus-B19-associated active myocarditis with biventricular thrombi. Results of endomyocardial biopsy investigations and cardiac magnetic resonance imaging. PMID- 17404166 TI - Images in cardiovascular medicine. Vanishing tumor. PMID- 17404167 TI - Letter by Conti and Volpe regarding article, "Cardiac troponin I but not cardiac troponin T induces severe autoimmune inflammation in the myocardium". PMID- 17404168 TI - The European Association of Percutaneous Cardiovascular Interventions. Interview with Dr. William Wijns by Emma Baines. PMID- 17404169 TI - The Fontan right atrium--in context. PMID- 17404170 TI - Cytomegalovirus infection and cardiac allograft vasculopathy in children. PMID- 17404171 TI - Metabolic syndrome: is it a syndrome? Does it matter? PMID- 17404172 TI - Metabolic syndrome: a work in progress, but a useful construct. PMID- 17404173 TI - Evaluation of randomized controlled trials. PMID- 17404175 TI - Upregulation of nitric oxide, inhibition of oxidative stress, and antihypertensive effects of statins. PMID- 17404178 TI - The riskiest time for the brain: could the nighttime be the right time for intervention? PMID- 17404179 TI - Linking cardiac mechanosensing at the sarcomere M-band, nuclear factor kappaB signaling, and cardiac remodeling. PMID- 17404180 TI - Sex differences in the renal changes elicited by angiotensin II blockade during the nephrogenic period. AB - The renin-angiotensin system plays an important role in renal development. However, it is unknown whether reduction in angiotensin II effects during the nephrogenic period leads to different renal alterations in males and females during the adult age. The aim of this study was to evaluate whether the role of angiotensin II on renal development is sex dependent and whether there are sex differences in blood pressure, renal hemodynamics, and severity of renal damage during adult life when nephrogenesis is altered by blocking angiotensin II effects. Newborn Sprague-Dawley rats were treated with an angiotensin II type 1 receptor antagonist (L-158.809; 7 mg/kg per day) during the first 2 weeks of life. At 3 months of age, changes in blood pressure, albuminuria, and renal hemodynamics were assessed, and stereological and histopathologic studies were performed. Blood pressure increased (127+/-0.5 versus 115+/-0.7 mm Hg in control rats; P<0.05) and nephron number decreased (37%; P<0.05) similarly in treated males and females. However, only males had an elevation in albuminuria (5.92+/ 1.65 versus 0.33+/-0.09 mg per day in control rats; P<0.05), a fall in glomerular filtration rate (12.6%; P<0.05), and a significant decrease in papillary volume (42%; P<0.05). Mean glomerular volume, glomerulosclerosis, arteriolar hypertrophy, and tubulointerstitial damage in cortex and medulla were also higher (P<0.05) in angiotensin II type 1 receptor antagonist-treated males than in treated females. The results of this study suggest that females seem to be more protected than males to the renal consequences of reducing angiotensin II effects during renal development. PMID- 17404181 TI - Left ventricular hypertrophy, subclinical atherosclerosis, and inflammation. AB - To elucidate mechanisms by which left ventricular (LV) hypertrophy (LVH) increases the risk of atherosclerotic heart disease, we sought to determine whether LVH is independently associated with coronary artery calcium (CAC) and serum C-reactive protein (CRP) levels in the general population. The Dallas Heart Study is a population-based sample in which 2633 individuals underwent cardiac MRI to measure LV structure, electron beam CT to measure CAC, and measurement of plasma CRP. We used univariate and multivariable analyses to determine whether LV mass and markers of concentric LV hypertrophy or dilation were associated with CAC and CRP. Increasing quartiles of LV mass indexed to fat-free mass, LV wall thickness, and concentricity, but not LV volume, were associated with CAC in both men and women (P<0.001). After adjustment for traditional cardiovascular risk factors and statin use, LV wall thickness and concentricity remained associated with CAC in linear regression (P<0.001 for each). These associations were particularly robust in blacks. LV wall thickness and concentricity were also associated with elevated CRP levels (P=0.001 for both) in gender-stratified univariate analyses, although these associations did not persist in multivariable analysis. In conclusion, concentric LVH is an independent risk factor for subclinical atherosclerosis. LVH is also associated with an inflammatory state as reflected in elevated CRP levels, although this relationship appears to be mediated by comorbid conditions. These data likely explain in part why individuals with LVH are at increased risk for myocardial infarction. PMID- 17404182 TI - Impedance progress: aortic diameter rears its head again? PMID- 17404183 TI - Noninvasive (input) impedance, pulse wave velocity, and wave reflection in healthy middle-aged men and women. AB - The relation between arterial function indices, such as pulse wave velocity and augmentation index with parameters derived from input impedance analysis, is still incompletely understood. Carotid pressure, central flow waveforms, and pulse wave velocity were noninvasively acquired in 2026 apparently healthy, middle-aged subjects (1052 women and 974 men) 35 to 55 years old at inclusion. Input and characteristic impedance, reflection coefficient, the ratio of backward to-forward pressure amplitude (reflection magnitude), and augmentation index were derived. Pulse wave velocity increased by 15% (from 6.1 to 7.0 m/s) both in men and women. In qualitative terms, input impedance evolved from a pattern indicative of wave transmission and reflection to a pattern more compatible with a windkessel-like system. In women, a decrease in total arterial compliance led to an increased input impedance in the low frequency range, whereas few changes were observed in men. Characteristic impedance did not change with age in women and even decreased in men (P<0.001) and could not be identified as the primary determinant of central pulse pressure. Augmentation index increased with age, as was expected, and was systematically higher in women (P<0.001). Reflection coefficient and reflection magnitude increased with age (P<0.001) without gender differences. We conclude that, in healthy middle-aged subjects, the age-related increase in arterial stiffness (pulse wave velocity) is not fully paralleled by an increase in arterial impedance, suggesting a role for age-dependent modulation of aortic cross-sectional area. Wave reflection increases with age and is not higher in women than in men. PMID- 17404184 TI - Similar associations of parental prenatal smoking suggest child blood pressure is not influenced by intrauterine effects. AB - Maternal smoking in pregnancy may be associated with higher offspring blood pressure; however, results of previous studies have been inconsistent and included varying confounder adjustments. We studied the association between maternal smoking in pregnancy and offspring blood pressure at 7 years in the Avon Longitudinal Study of Parents and Children, accounting for important social and environmental confounders and using partner smoking to investigate intrauterine effects. Analysis was carried out in 6509 children with maternal smoking data and 7149 children with partner smoking data. In models adjusting for child age and sex, modest differences in systolic blood pressure were observed between children of mothers who did and did not smoke during pregnancy (beta=0.64 mm Hg; 95% CI: 0.09 to 1.20; P=0.02). Adjusting for all of the confounders attenuated this difference toward the null (beta=0.05 mm Hg; 95% CI: -0.59 to 0.68; P=0.9), mostly because of adjustment for breastfeeding, maternal education, and family social class. Associations were similar between maternal and partner smoking with offspring systolic blood pressure (for partner smoking: beta=0.62 mm Hg; 95% CI: 0.17 to 1.07; P=0.07 minimally adjusted and beta=0.26 mm Hg; 95% CI: -0.36 to 0.87; P=0.4 fully adjusted), providing further evidence that differences in child blood pressure observed in minimally adjusted models are not because of a biological influence of maternal smoking on the intrauterine environment. PMID- 17404185 TI - Vascular stiffness and genetic variation at the endothelial nitric oxide synthase locus: the Framingham Heart study. AB - Arterial stiffness is a moderately heritable trait that is affected by alterations in the bioavailability of NO. Previous studies have found associations between variants in the gene for endothelial NO synthase (NOS3) and arterial properties. We previously identified a linkage peak for forward pressure wave amplitude in the immediate vicinity of NOS3. Therefore, we evaluated relations between arterial stiffness measures and common genetic variants at this locus. Eighteen single nucleotide polymorphisms capturing approximately 90% of underlying common variation in NOS3 were genotyped in unrelated Framingham Heart Study participants (N=1157; 52.2% women; mean age: 62 years) with routinely ascertained tonometry data that provided 5 arterial phenotypes (forward and reflected pressure wave amplitude, central pulse pressure, carotid-femoral pulse wave velocity, and mean arterial pressure). In women but not men, the genotype for the common NOS3 missense mutation (Glu298Asp, rs1799983) was related to central pulse pressure (women: GG=53+/-0.9, GT=54+/-0.9, and TT=47+/-2.0 mm Hg, P=0.0047; men: GG=50+/-1.0, GT=49+/-0.9, and TT=47+/-1.8 mm Hg, P=0.30) and forward wave amplitude (women: GG=41+/-0.7, GT=42+/-0.7, and TT=38+/-1.6 mm Hg, P=0.029; men: GG=42+/-0.9, GT=41+/-0.8, and TT=39+/-1.5 mm Hg, P=0.47). The only other nominally significant sex-specific association in men but not women was between an intronic polymorphism (rs1800781) and reflected wave amplitude (women: AA=10.4+/-0.4, AG=11.1+/-0.6, and GG=8.9+/-2.2 mm Hg, P=0.50; men: AA=6.1+/-0.3, AG=7.3+/-0.5, and GG=11.3+/-2.3 mm Hg, P=0.014). After adjusting for multiple testing (18 polymorphisms and 5 phenotypes), these nominal associations were no longer significant. The present study was suggestive of modest relations between common genetic variants at the NOS3 locus and arterial stiffness. PMID- 17404186 TI - Regulation of scavenger receptor class BI gene expression by angiotensin II in vascular endothelial cells. AB - High-density lipoprotein mediates a normal physiological process called reverse cholesterol transport. In this process, a scavenger receptor of the B class (SR BI)/human homologue of SR-BI, CD36, and LIMPII analogous-1 (hSR-BI/CLA-1) facilitates the cellular uptake of cholesterol from high-density lipoprotein. In endothelial cells, high-density lipoprotein activates endothelial NO synthase via hSR-BI/CLA-1. Angiotensin II (Ang II) is a powerful accelerator of atherosclerosis and modulates the expression of endothelial NO synthase. In the present study, we have examined the role of Ang II on hSR-BI/CLA-1 expression in human umbilical vein endothelial cells. Our results showed that endogenous expression of hSR-BI/CLA-1 was suppressed by exposure to Ang II in human umbilical vein endothelial cells. Administration of the Ang II type-1 receptor blocker olmesartan inhibited Ang II-induced hSR-BI/CLA-1 protein repression. In Ang II-treated cells, high-density lipoprotein had no effect on endothelial NO synthase activation. Ang II decreased transcriptional activity of the hSR-BI/CLA 1 promoter. The inhibitory effect of Ang II on hSR-BI/CLA-1 promoter activity was abrogated by wortmannin and LY294002, specific inhibitors of phosphatidylinositol 3-kinase. Exposure of human umbilical vein endothelial cells to Ang II elicited a rapid phosphorylation of Akt and FoxO1, a known target of Akt signaling. Constitutively active Akt inhibits the activity of the hSR-BI/CLA-1 promoter, and a dominant-negative mutant of Akt or mutagenesis of a FoxO1 response element in the hSR-BI/CLA-1 abolished the ability of Ang II to suppress promoter activity. Together, these results indicate that the phosphatidylinositol 3-kinase/Akt/FoxO1 pathway participates in Ang II suppression of hSR-BI/CLA-1 expression and suggests that the endothelial receptor for hSR-BI/CLA-1 is downregulated by the renin-angiotensin system. PMID- 17404187 TI - Barbershops as hypertension detection, referral, and follow-up centers for black men. AB - Barbershops constitute potential sites for community health promotion programs targeting hypertension (HTN) in black men, but such programs have not been evaluated previously. Here we conducted 2 nonrandomized feasibility studies to determine whether an enhanced intervention program of continuous blood pressure (BP) monitoring and peer-based health messaging in a barbershop lowers BP more than standard screening and health education (study 1) and can be implemented by barbers rather than research personnel (study 2). In study 1, we measured changes in HTN treatment and BP in regular barbershop customers with poorly controlled HTN assigned for 8 months to either an enhanced intervention group (n=36) or a contemporaneous comparison group (n=27). Groups were similar at baseline. BP fell by 16+/-3/9+/-2 mm Hg in the enhanced intervention group but was unchanged in the comparison group (P<0.0001, adjusted for age and body mass index). HTN treatment and control increased from 47% to 92% (P<0.001) and 19% to 58% (P<0.001), respectively, in the enhanced intervention group, whereas both remained unchanged in the comparison group. In study 2, barbers were trained to administer the enhanced intervention continuously for 14 months to the entire adult black male clientele (n=321) in 1 shop. Six barbers recorded 8953 BP checks during 11 066 haircuts, thus demonstrating a high degree of intervention fidelity. Furthermore, among 107 regular customers with HTN, treatment and control increased progressively with increasing intervention exposure (P<0.01). Taken together, these data suggest that black-owned barbershops can be transformed into effective HTN detection, referral, and follow-up centers. Further research is warranted. PMID- 17404188 TI - Association of ambulatory blood pressure with ischemic brain injury. AB - Cerebral white matter hyperintensities on brain MRI (leukoaraiosis) are associated with increased risk of stroke and dementia. To assess the relationships of blood pressure level and circadian pattern with leukoaraiosis, we obtained 24-hour ambulatory blood pressure recordings and brain magnetic resonance images in 343 white and 267 black adults who were members of sibships that had >or=2 siblings with essential hypertension. In multiple linear regression models, factors associated with greater leukoaraiosis in both racial groups included age (P15 kDa in size, Nuclear Overhauser effect-derived distance restraints are difficult to obtain, and RDCs could serve as the main reliable source of NMR structural information. In many of these cases, RDCs would enable full structure determination that otherwise would be impossible. However, none of the existing liquid-crystalline media used to align water-soluble proteins are compatible with the detergents required to solubilize membrane proteins. We report the design and construction of a detergent-resistant liquid crystal of 0.8-microm-long DNA-nanotubes that can be used to induce weak alignment of membrane proteins. The nanotubes are heterodimers of 0.4-microm-long six-helix bundles each self-assembled from a 7.3 kb scaffold strand and >170 short oligonucleotide staple strands. We show that the DNA-nanotube liquid crystal enables the accurate measurement of backbone N(H) and C(alpha)H(alpha) RDCs for the detergent-reconstituted zeta-zeta transmembrane domain of the T cell receptor. The measured RDCs validate the high-resolution structure of this transmembrane dimer. We anticipate that this medium will extend the advantages of weak alignment to NMR structure determination of a broad range of detergent-solubilized membrane proteins. PMID- 17404218 TI - Regulation of late B cell differentiation by intrinsic IKKalpha-dependent signals. AB - NF-kappaB-inducing kinase (NIK)-mediated IKKalpha phosphorylation activates the alternative NF-kappaB pathway, which is characterized by nuclear translocation of p52:RelB heterodimers. This alternative pathway is initiated by a select few receptors, including LT-betaR, BAFF-R, and CD40. Although NIK, IKKalpha, and p52 are all critical regulators of LT-betaR signaling in stromal cells during humoral immune responses, lymphocytes require NIK, but not p52, for optimal Ig production. This disparity suggests that NIK possesses critical cell-type specific functions that do not depend on NF-kappaB. Here we use mice bearing targeted mutations of the IKKalpha activation loop Ser(176/180) (IKKalpha(AA)) to address the B cell-intrinsic functions of NIK-IKKalpha signaling in vivo. We find that IKKalpha(AA) B cells mount normal primary antibody responses but do not enter germinal centers. This defect likely derives from ineffective early T-B cell collaboration and leads to impaired generation of humoral memory and relatively short-lived, low-affinity antibody production. Our findings contrast with those obtained by using p52(-/-) B cells, which mount normal Ig responses, and alymphoplasia (NIK mutant) B cells, which produce very little primary Ig. Thus, the NIK-IKKalpha-p52 axis is not as linear and exclusive as previous studies suggest, and IKKalpha possesses critical NF-kappaB-independent functions in B cells. PMID- 17404219 TI - Phosphoproteomic identification of targets of the Arabidopsis sucrose nonfermenting-like kinase SnRK2.8 reveals a connection to metabolic processes. AB - SnRK2.8 is a member of the sucrose nonfermenting-related kinase family that is down-regulated when plants are deprived of nutrients and growth is reduced. When this kinase is over expressed in Arabidopsis, the plants grow larger. To understand how this kinase modulates growth, we identified some of the proteins that are phosphorylated by this kinase. A new phosphoproteomic method was used in which total protein from plants overexpressing the kinase was compared with total protein from plants in which the kinase was inactivated. Protein profiles were compared on two-dimensional gels following staining by a dye that recognizes phosphorylated amino acids. Candidate target proteins were confirmed with in vitro phosphorylation assays, using the kinase and target proteins that were purified from Escherichia coli. Seven target proteins were confirmed as being phosphorylated by SnRK2.8. Certain targets, such as 14-3-3 proteins, regulate as yet unidentified proteins, whereas other targets, such as glyoxalase I and ribose 5-phosphate isomerase, detoxify byproducts from glycolysis and catalyze one of the final steps in carbon fixation, respectively. Also, adenosine kinase and 60S ribosomal protein were confirmed as targets of SnRK2.8. Using mass spectrometry, we identified phosphorylated residues in the SnRK2.8, the 14-3-3kappa, and the 14 3-3chi. These data show that the expression of SnRK2.8 is correlated with plant growth, which may in part be due to the phosphorylation of enzymes involved in metabolic processes. PMID- 17404220 TI - Evidence that cochlear-implanted deaf patients are better multisensory integrators. AB - The cochlear implant (CI) is a neuroprosthesis that allows profoundly deaf patients to recover speech intelligibility. This recovery goes through long-term adaptative processes to build coherent percepts from the coarse information delivered by the implant. Here we analyzed the longitudinal postimplantation evolution of word recognition in a large sample of CI users in unisensory (visual or auditory) and bisensory (visuoauditory) conditions. We found that, despite considerable recovery of auditory performance during the first year postimplantation, CI patients maintain a much higher level of word recognition in speechreading conditions compared with normally hearing subjects, even several years after implantation. Consequently, we show that CI users present higher visuoauditory performance when compared with normally hearing subjects with similar auditory stimuli. This better performance is not only due to greater speechreading performance, but, most importantly, also due to a greater capacity to integrate visual input with the distorted speech signal. Our results suggest that these behavioral changes in CI users might be mediated by a reorganization of the cortical network involved in speech recognition that favors a more specific involvement of visual areas. Furthermore, they provide crucial indications to guide the rehabilitation of CI patients by using visually oriented therapeutic strategies. PMID- 17404221 TI - Composite synthetic lethal identification of membrane traffic inhibitors. AB - Small molecule inhibitors provide powerful tools to characterize highly dynamic and complex eukaryotic cell pathways such as those mediating membrane traffic. However, a lack of easy and generalizable assays has constrained identification of novel inhibitors despite availability of diverse chemical libraries. Here, we report a facile growth-based strategy in yeast to screen for pathway-specific inhibitors. The approach uses well characterized synthetic genetic growth defects to guide design of cells genetically sensitized for inhibition of chosen pathways. With this strategy, we identified a family of piperazinyl phenylethanone compounds as inhibitors of traffic between the trans-Golgi network (TGN) and endosomes that depends on the clathrin adaptor complex AP-1. The compounds did not significantly alter other trafficking pathways involving the TGN or endosomes, indicating specificity. Compound treatment also altered localization of AP-1 in mammalian cells. These previously uncharacterized inhibitors will be useful for future studies of clathrin-mediated transport in yeast, and potentially in other organisms. Furthermore, the easily automated technology should be adaptable for identification of inhibitors of other cellular processes. PMID- 17404222 TI - Rat Mcs5a is a compound quantitative trait locus with orthologous human loci that associate with breast cancer risk. AB - Breast cancer risk is a polygenic trait. To identify breast cancer modifier alleles that have a high population frequency and low penetrance we used a comparative genomics approach. Quantitative trait loci (QTL) were initially identified by linkage analysis in a rat mammary carcinogenesis model followed by verification in congenic rats carrying the specific QTL allele under study. The Mcs5a locus was identified by fine-mapping Mcs5 in a congenic model. Here we characterize the Mcs5a locus, which when homozygous for the Wky allele, reduces mammary cancer risk by 50%. The Mcs5a locus is a compound QTL with at least two noncoding interacting elements: Mcs5a1 and Mcs5a2. The resistance phenotype is only observed in rats carrying at least one copy of the Wky allele of each element on the same chromosome. Mcs5a1 is located within the ubiquitin ligase Fbxo10, whereas Mcs5a2 includes the 5' portion of Frmpd1. Resistant congenic rats show a down-regulation of Fbxo10 in the thymus and an up-regulation of Frmpd1 in the spleen. The association of the Mcs5a1 and Mcs5a2 human orthologs with breast cancer was tested in two population-based breast cancer case-control studies (approximately 12,000 women). The minor alleles of rs6476643 (MCS5A1) and rs2182317 (MCS5A2) were independently associated with breast cancer risk. The minor allele of rs6476643 increases risk, whereas the rs2182317 minor allele decreases risk. Both alleles have a high population frequency and a low penetrance toward breast cancer risk. PMID- 17404223 TI - The role of CaMKII as an F-actin-bundling protein crucial for maintenance of dendritic spine structure. AB - Ca(2+)-calmodulin-dependent protein kinase II (CaMKII) is a serine/threonine protein kinase critically involved in synaptic plasticity in the brain. It is highly concentrated in the postsynaptic density fraction, exceeding the amount of any other signal transduction molecules. Because kinase signaling can be amplified by catalytic reaction, why CaMKII exists in such a large quantity has been a mystery. Here, we provide biochemical evidence that CaMKII is capable of bundling F-actin through a stoichiometric interaction. Consistent with this evidence, in hippocampal neurons, RNAi-mediated down-regulation of CaMKII leads to a reduction in the volume of dendritic spine head that is mediated by F-actin dynamics. An overexpression of CaMKII slowed down the actin turnover in the spine head. This activity was associated with beta subunit of CaMKII in a manner requiring its actin-binding and association domains but not the kinase domain. This finding indicates that CaMKII serves as a central signaling molecule in both functional and structural changes during synaptic plasticity. PMID- 17404224 TI - Evolution and genetic differentiation among relatives of Arabidopsis thaliana. AB - Arabidopsis thaliana is one of the most intensively studied plant species. More recently, information is accumulating about its closest relatives, the former genus Cardaminopsis. A. thaliana diverged from these relatives, actually treated within three major lineages (Arabidopsis lyrata, Arabidopsis halleri, and Arabidopsis arenosa), approximately 5 mya. Significant karyotype evolution in A. thaliana with base chromosome number reduction from x=8 to x=5 might indicate and favor effective genetic isolation from these other species, although hybrids are occurring naturally and have been also constituted under controlled conditions. We tested the evolutionary significance to separate the x=5 from the x=8 lineage using DNA sequence data from the plastome and the nuclear ribosomal DNA based on an extensive, representative worldwide sampling of nearly all taxonomic entities. We conclude that (i) A. thaliana is clearly separated phylogenetically from the x=8 lineage, (ii) five major lineages outside A. thaliana can be identified (A. lyrata, A. arenosa, A. halleri, Arabidopsis croatica, and Arabidopsis pedemontana) together with Arabidopsis cebennensis, and (iii) centers of genetic and morphological diversity are mostly in congruence and are located close to the Balkans in Austria and Slovakia outside glaciated and permafrost regions with few notable exceptions. PMID- 17404225 TI - Protein identification by spectral networks analysis. AB - Advances in tandem mass spectrometry (MS/MS) steadily increase the rate of generation of MS/MS spectra. As a result, the existing approaches that compare spectra against databases are already facing a bottleneck, particularly when interpreting spectra of modified peptides. Here we explore a concept that allows one to perform an MS/MS database search without ever comparing a spectrum against a database. We propose to take advantage of spectral pairs, which are pairs of spectra obtained from overlapping (often nontryptic) peptides or from unmodified and modified versions of the same peptide. Having a spectrum of a modified peptide paired with a spectrum of an unmodified peptide allows one to separate the prefix and suffix ladders, to greatly reduce the number of noise peaks, and to generate a small number of peptide reconstructions that are likely to contain the correct one. The MS/MS database search is thus reduced to extremely fast pattern-matching (rather than time-consuming matching of spectra against databases). In addition to speed, our approach provides a unique paradigm for identifying posttranslational modifications by means of spectral networks analysis. PMID- 17404226 TI - In vivo CD8+ T cell control of immunodeficiency virus infection in humans and macaques. AB - Forty million people are estimated to be infected with HIV-1, and only a small fraction of those have access to life-prolonging antiretroviral treatment. As the epidemic grows there is an urgent need for effective therapeutic and prophylactic vaccines. Nonhuman primate models of immunodeficiency virus infection are essential for the preclinical evaluation of candidate vaccines. To interpret the results of these trials, comparative studies of the human and macaque immune responses are needed. Despite the widespread use of macaques to evaluate vaccines designed to elicit a CD8(+) cytotoxic T lymphocyte (CTL) response, the efficiency with which CTL control immunodeficiency virus infections has not been compared between humans and macaques, largely because of difficulties in assaying the functional CTL response. We recently developed a method for estimating the rate at which CTLs kill cells productively infected with HIV-1 in humans in vivo. Here, using the same technique, we quantify the rate at which CTLs kill infected cells in macaque models of HIV infection. We show that CTLs kill productively infected cells significantly faster (P = 0.004) and that escape variants have significantly higher fitness costs (P = 0.003) in macaques compared with humans. These results suggest that it may be easier to elicit a protective CTL response in macaques than in humans and that vaccine studies conducted in macaques need to be interpreted accordingly. PMID- 17404227 TI - Reduced expression of mitochondrial frataxin in mice exacerbates diet-induced obesity. AB - Published evidence suggests that adiposity in humans may be linked to impaired energy expenditure for reasons widely unresolved. We have generated mice with a systemic impairment of oxidative phosphorylation (OXPHOS) due to aP2 cre-mediated targeted disruption, and unexpectedly ubiquitous reduction of mitochondrial frataxin protein expression. Only when maintained on a high-calorie diet resembling Westernized eating habits, these animals accumulate additional body fat, leading to increased body mass, and develop diabetes mellitus, despite the fact that both calorie uptake and physical activity were identical to that in control animals. This phenotype is caused by a mild but significant reduction in total energy expenditure paralleled by increased expression of ATP citrate lyase, a rate-limiting step in de novo synthesis of fatty acids and triglycerides. Taken together, these findings indicate that a limited impairment in oxidative metabolism within the mitochondria directly predisposes mammals to excessive body weight gain. PMID- 17404228 TI - Cryptic proteolytic activity of dihydrolipoamide dehydrogenase. AB - The mitochondrial enzyme, dihydrolipoamide dehydrogenase (DLD), is essential for energy metabolism across eukaryotes. Here, conditions known to destabilize the DLD homodimer enabled the mouse, pig, or human enzyme to function as a protease. A catalytic dyad (S456-E431) buried at the homodimer interface was identified. Serine protease inhibitors and an S456A or an E431A point mutation abolished the proteolytic activity, whereas other point mutations at the homodimer interface domain enhanced the proteolytic activity, causing partial or complete loss of DLD activity. In humans, mutations in the DLD homodimer interface have been linked to an atypical form of DLD deficiency. These findings reveal a previously unrecognized mechanism by which certain DLD mutations can simultaneously induce the loss of a primary metabolic activity and the gain of a moonlighting proteolytic activity. The latter could contribute to the metabolic derangement associated with DLD deficiency and represent a target for therapies of this condition. PMID- 17404229 TI - Rubisco large-subunit translation is autoregulated in response to its assembly state in tobacco chloroplasts. AB - Plants rely on ribulose bisphosphate carboxylase/oxygenase (Rubisco) for carbon fixation. Higher plant Rubisco possesses an L(8)S(8) structure, with the large subunit (LS) encoded in the chloroplast by rbcL and the small subunit encoded by the nuclear RBCS gene family. Because its components accumulate stoichiometrically but are encoded in two genetic compartments, rbcL and RBCS expression must be tightly coordinated. Although this coordination has been observed, the underlying mechanisms have not been defined. Here, we use tobacco to understand how LS translation is related to its assembly status. To do so, two transgenic lines deficient in LS biogenesis were created: a chloroplast transformant expressing a truncated and unstable LS polypeptide, and a line where a homolog of the maize Rubisco-specific chaperone, BSD2, was repressed by RNAi. We found that in both lines, LS translation is no longer regulated by the availability of small subunit (SS), indicating that LS translation is not activated by the presence of its assembly partner but, rather, undergoes an autoregulation of translation. Pulse labeling experiments indicate that LS is synthesized but not accumulated in the transgenic lines, suggesting that accumulation of a repressor motif is required for LS assembly-dependent translational regulation. PMID- 17404230 TI - Subversion of B lymphocyte tolerance by hydralazine, a potential mechanism for drug-induced lupus. AB - Accumulating evidence indicates that epigenetic alterations contribute to exacerbated activation or deregulation of the mechanisms that maintain tolerance to self-antigens in patients with lupus, a systemic autoimmune disease that can be triggered by medications taken to treat a variety of conditions. Here, we tested the effect of hydralazine, an antihypertensive drug that triggers lupus, on receptor editing, a chief mechanism of B lymphocyte tolerance to self antigens. Using mice expressing transgenic human Igs, we found that hydralazine impairs up-regulation of RAG-2 gene expression and reduces secondary Ig gene rearrangements. Receptor editing was also partially abolished in a dose-dependent manner by a specific inhibitor of MEK1/2. Adoptive transfer of bone marrow B cells pretreated with hydralazine or with a MEK inhibitor to naive syngeneic mice resulted in autoantibody production. We conclude that, by disrupting receptor editing, hydralazine subverts B lymphocyte tolerance to self and contributes to generation of pathogenic autoreactivity. We also postulate that inhibition of the Erk signaling pathway contributes to the pathogenesis of hydralazine-induced lupus and idiopathic human lupus. PMID- 17404231 TI - Regulation of ryanodine receptor-dependent calcium signaling by polycystin-2. AB - Mutations in polycystin-2 (PC2) cause autosomal dominant polycystic kidney disease. A function for PC2 in the heart has not been described. Here, we show that PC2 coimmunoprecipitates with the cardiac ryanodine receptor (RyR2) from mouse heart. Biochemical assays showed that the N terminus of PC2 binds the RyR2, whereas the C terminus only binds to RyR2 in its open state. Lipid bilayer electrophysiological experiments indicated that the C terminus of PC2 functionally inhibited RyR2 channel activity in the presence of calcium (Ca(2+)). Pkd2(-/-) cardiomyocytes had a higher frequency of spontaneous Ca(2+) oscillations, reduced Ca(2+) release from the sarcoplasmic reticulum stores, and reduced Ca(2+) content compared with Pkd2(+/+) cardiomyocytes. In the presence of caffeine, Pkd2(-/-) cardiomyocytes exhibited decreased peak fluorescence, a slower rate of rise, and a longer duration of Ca(2+) transients compared with Pkd2(+/+). These data suggest that PC2 is important for regulation of RyR2 function and that loss of this regulation of RyR2, as occurs when PC2 is mutated, results in altered Ca(2+) signaling in the heart. PMID- 17404232 TI - Identification of a bone marrow precursor of the earliest thymocytes in adult mouse. AB - The thymus requires continuous replenishment of progenitors from the bone marrow (BM) to sustain T cell development. However, it remains unclear which hematopoietic progenitors downstream from hematopoietic stem cells in the BM home to the thymus in adult mice. In this work, we demonstrate that although multiple BM populations have intrinsic T lineage differentiation potential, a small subset of multipotent progenitors (MPPs) expressing CCR9 preferentially homes to the thymus. These CCR9(+) MPPs are phenotypically similar to the most immature early T lineage progenitors (ETPs) in the thymus and are present in the peripheral blood. Similar to ETPs, CCR9(+) MPPs undergo Notch signaling, as indicated by higher expression of Notch1 and downstream target Hes1 genes compared with other MPP subsets. Furthermore, CCR9(+) MPPs possess differentiation potential similar to that of ETPs, with very limited granulocyte/macrophage differentiation potential, but they can differentiate into T, B, and dendritic cells. These characteristics implicate CCR9(+) MPPs as the BM precursors of the earliest thymic progenitors. In addition, our data suggest that before transition from BM to thymus, MPPs are lymphoid-specified and primed for T lineage differentiation. PMID- 17404233 TI - CCR9 is a homing receptor for plasmacytoid dendritic cells to the small intestine. AB - Small intestine plasmacytoid dendritic cells (pDC) are poorly characterized. Here, we demonstrate that intestinal pDC show the characteristic plasma cell-like morphology, and are recognized by antibodies against B220, Ly6c, 120G8, and PDCA 1, markers that are typically expressed by pDC. Furthermore, intestinal pDC carry high levels of CCR9 and are largely absent in the intestine, but not in lung, liver, or secondary lymphoid organs of CCR9-deficient animals. Competitive adoptive transfers reveal that CCR9-deficient pDC are impaired in homing to the small intestine after i.v. transfer. In a model of cholera toxin-induced gut inflammation, pDC are recruited to the intestine in WT but not CCR9-deficient animals. Furthermore, after oral application of a Toll-like receptor (TLR) 7/8 ligand, myeloid DC of the lamina propria are rapidly mobilized in WT but not in CCR9-deficient animals. Mobilization of myeloid DC can be completely rescued by adoptively transferred WT pDC to CCR9-deficient mice before oral challenge. Together, our data reveal an essential role for CCR9 in the homing of pDC to the intestine under homeostatic and inflammatory conditions and demonstrate an important role for intestinal pDC for the rapid mobilization of lamina propria DC. PMID- 17404234 TI - X-ray structure analysis of a metalloprotein with enhanced active-site resolution using in situ x-ray absorption near edge structure spectroscopy. AB - X-ray absorption spectroscopy is exquisitely sensitive to the coordination geometry of an absorbing atom and therefore allows bond distances and angles of the surrounding atomic cluster to be measured with atomic resolution. By contrast, the accuracy and resolution of metalloprotein active sites obtainable from x-ray crystallography are often insufficient to analyze the electronic properties of the metals that are essential for their biological functions. Here, we demonstrate that the combination of both methods on the same metalloprotein single crystal yields a structural model of the protein with exceptional active site resolution. To this end, we have collected an x-ray diffraction data set to 1.4-A resolution and Fe K-edge polarized x-ray absorption near edge structure (XANES) spectra on the same cyanomet sperm whale myoglobin crystal. The XANES spectra were quantitatively analyzed by using a method based on the multiple scattering approach, which yielded Fe-heme structural parameters with +/-(0.02 0.07)-A accuracy on the atomic distances and +/-7 degrees on the Fe-CN angle. These XANES-derived parameters were subsequently used as restraints in the crystal structure refinement. By combining XANES and x-ray diffraction, we have obtained an cyanomet sperm whale myoglobin structural model with a higher precision of the bond lengths and angles at the active site than would have been possible with crystallographic analysis alone. PMID- 17404235 TI - Behavioral changes induced by Toxoplasma infection of rodents are highly specific to aversion of cat odors. AB - The protozoan parasite Toxoplasma gondii blocks the innate aversion of rats for cat urine, instead producing an attraction to the pheromone; this may increase the likelihood of a cat predating a rat. This is thought to reflect adaptive, behavioral manipulation by Toxoplasma in that the parasite, although capable of infecting rats, reproduces sexually only in the gut of the cat. The "behavioral manipulation" hypothesis postulates that a parasite will specifically manipulate host behaviors essential for enhancing its own transmission. However, the neural circuits implicated in innate fear, anxiety, and learned fear all overlap considerably, raising the possibility that Toxoplasma may disrupt all of these nonspecifically. We investigated these conflicting predictions. In mice and rats, latent Toxoplasma infection converted the aversion to feline odors into attraction. Such loss of fear is remarkably specific, because infection did not diminish learned fear, anxiety-like behavior, olfaction, or nonaversive learning. These effects are associated with a tendency for parasite cysts to be more abundant in amygdalar structures than those found in other regions of the brain. By closely examining other types of behavioral patterns that were predicted to be altered we show that the behavioral effect of chronic Toxoplasma infection is highly specific. Overall, this study provides a strong argument in support of the behavioral manipulation hypothesis. Proximate mechanisms of such behavioral manipulations remain unknown, although a subtle tropism on part of the parasite remains a potent possibility. PMID- 17404236 TI - Hydrophobic association of alpha-helices, steric dewetting, and enthalpic barriers to protein folding. AB - Efficient protein folding implies a microscopic funnel-like multidimensional free energy landscape. Macroscopically, conformational entropy reduction can manifest itself as part of an empirical barrier in the traditional view of folding, but experiments show that such barriers can also entail significant unfavorable enthalpy changes. This observation raises the puzzling possibility, irrespective of conformational entropy, that individual microscopic folding trajectories may encounter large uphill moves and thus the multidimensional free-energy landscape may not be funnel-like. Here, we investigate how nanoscale hydrophobic interactions might underpin this salient enthalpic effect in biomolecular assembly by computer simulations of the association of two preformed polyalanine or polyleucine helices in water. We observe a high, positive enthalpic signature at room temperature when the helix separation is less than a single layer of water molecules. Remarkably, this unfavorable enthalpy change, with a parallel increase in void volume, is largely compensated for by a concomitant increase in solvent entropy, netting only a small or nonexistent microscopic free-energy barrier. Thus, our findings suggest that high enthalpic folding barriers can be consistent with a funnel picture of folding and are mainly a desolvation phenomenon indicative of a cooperative mechanism of simultaneous formation of multiple side-chain contacts at the rate-limiting step. PMID- 17404237 TI - Lipoprotein SmpA is a component of the YaeT complex that assembles outer membrane proteins in Escherichia coli. AB - A major role of the outer membrane (OM) of Gram-negative bacteria is to provide a protective permeability barrier for the cell, and proper maintenance of the OM is required for cellular viability. OM biogenesis requires the coordinated assembly of constituent lipids and proteins via dedicated OM assembly machineries. We have previously shown that, in Escherichia coli, the multicomponent YaeT complex is responsible for the assembly of OM beta-barrel proteins (OMPs). This complex contains the OMP YaeT and three OM lipoproteins. Here, we report another component of the YaeT complex, the OM lipoprotein small protein A (SmpA). Strains carrying loss-of-function mutations in smpA are viable but exhibit defects in OMP assembly. Biochemical experiments show that SmpA is involved in maintaining complex stability. Taken together, these experiments establish an important role for SmpA in both the structure and function of the YaeT complex. PMID- 17404238 TI - Wnt signaling regulates pancreatic beta cell proliferation. AB - There is widespread interest in defining factors and mechanisms that stimulate proliferation of pancreatic islet cells. Wnt signaling is an important regulator of organ growth and cell fates, and genes encoding Wnt-signaling factors are expressed in the pancreas. However, it is unclear whether Wnt signaling regulates pancreatic islet proliferation and differentiation. Here we provide evidence that Wnt signaling stimulates islet beta cell proliferation. The addition of purified Wnt3a protein to cultured beta cells or islets promoted expression of Pitx2, a direct target of Wnt signaling, and Cyclin D2, an essential regulator of beta cell cycle progression, and led to increased beta cell proliferation in vitro. Conditional pancreatic beta cell expression of activated beta-catenin, a crucial Wnt signal transduction protein, produced similar phenotypes in vivo, leading to beta cell expansion, increased insulin production and serum levels, and enhanced glucose handling. Conditional beta cell expression of Axin, a potent negative regulator of Wnt signaling, led to reduced Pitx2 and Cyclin D2 expression by beta cells, resulting in reduced neonatal beta cell expansion and mass and impaired glucose tolerance. Thus, Wnt signaling is both necessary and sufficient for islet beta cell proliferation, and our study provides previously unrecognized evidence of a mechanism governing endocrine pancreas growth and function. PMID- 17404239 TI - Competing ferromagnetism in high-temperature copper oxide superconductors. AB - The extreme variability of observables across the phase diagram of the cuprate high-temperature superconductors has remained a profound mystery, with no convincing explanation for the superconducting dome. Although much attention has been paid to the underdoped regime of the hole-doped cuprates because of its proximity to a complex Mott insulating phase, little attention has been paid to the overdoped regime. Experiments are beginning to reveal that the phenomenology of the overdoped regime is just as puzzling. For example, the electrons appear to form a Landau Fermi liquid, but this interpretation is problematic; any trace of Mott phenomena, as signified by incommensurate antiferromagnetic fluctuations, is absent, and the uniform spin susceptibility shows a ferromagnetic upturn. Here, we show and justify that many of these puzzles can be resolved if we assume that competing ferromagnetic fluctuations are simultaneously present with superconductivity, and the termination of the superconducting dome in the overdoped regime marks a quantum critical point beyond which there should be a genuine ferromagnetic phase at zero temperature. We propose experiments and make predictions to test our theory and suggest that an effort must be mounted to elucidate the nature of the overdoped regime, if the problem of high-temperature superconductivity is to be solved. Our approach places competing order as the root of the complexity of the cuprate phase diagram. PMID- 17404240 TI - Ubiquitin-conjugating enzyme Ubc13 is a critical component of TNF receptor associated factor (TRAF)-mediated inflammatory responses. AB - Ubc13 is a ubiquitin-conjugating enzyme responsible for noncanonical ubiquitination of TNF receptor-associated factor (TRAF)-family adapter proteins involved in Toll-like receptor and TNF-family cytokine receptor signaling, which are regulators of innate immunity. Gene ablation was used to study the function of Ubc13 in mice. Whereas homozygous ubc13 gene disruption resulted in embryonic lethality, heterozygous ubc13(+/-) mice appeared normal, without alterations in immune cell populations. Haploinsufficient ubc13(+/-) mice were resistant to lipopolysaccharide-induced lethality, and demonstrated reduced in vivo ubiquitination of TRAF6. Macrophages and splenocytes isolated from ubc13(+/-) mice exhibited reduced lipopolysaccharide-inducible cytokine secretion and impaired activation of TRAF-dependent signal transduction pathways (NF-kappaB, JNK, and p38 MAPK). These findings document a critical role for Ubc13 in inflammatory responses and suggest that agents reducing Ubc13 activity could have therapeutic utility. PMID- 17404241 TI - The role of lubricin in the mechanical behavior of synovial fluid. AB - Synovial fluid is a semidilute hyaluronate (HA) polymer solution, the rheology of which depends on HA-protein interactions, and lubricin is a HA-binding protein found in synovial fluid and at cartilage surfaces, where it contributes to boundary lubrication under load. Individuals with genetic deficiency of lubricin develop precocious joint failure. The role of lubricin in synovial fluid rheology is not known. We used a multiple-particle-tracking microrheology technique to study the molecular interactions between lubricin and HA in synovial fluid. Particles (200 nm mean diameter) embedded in normal and lubricin-deficient synovial fluid samples were tracked separately by using multiple-particle tracking microrheology. The time-dependent ensemble-averaged mean-squared displacements of all of the particles were measured over a range of physiologically relevant frequencies. The mean-squared displacement correlation with time lag had slopes with values of unity for simple HA solutions and for synovial fluid from an individual who genetically lacked lubricin, in contrast to slopes with values less than unity (alpha approximately 0.6) for normal synovial fluid. These data correlated with bulk rheology studies of the same samples. We found that the subdiffusive and elastic behavior of synovial fluid, at physiological shear rates, was absent in fluid from a patient who lacks lubricin. We conclude that lubricin provides synovial fluid with an ability to dissipate strain energy induced by mammalian locomotion, which is a chondroprotective feature that is distinct from boundary lubrication. PMID- 17404242 TI - Raman spectra of a Lower Cambrian ctenophore embryo from southwestern Shaanxi, China. AB - The Early Cambrian (approximately 540 million years old) Meishucun fossil assemblage of Ningqiang County (Shaanxi Province), China, contains the oldest complex skeletonized organisms known in the geological record. We here report the finding in this assemblage of an exquisitely preserved late-stage embryo of a ctenophore ("comb jelly"), its fine structure documented by confocal laser scanning microscopy and shown by Raman spectroscopy to be composed of carbonaceous kerogen permineralized in apatite. In its spheroidal morphology, the presence of eight comb rows and the absence of tentacles, this embryo resembles an adult ctenophore (Maotianoascus octonarius) known from the immediately younger Chengjiang fauna of Yunnan, China. The oldest ctenophore and the only embryonic comb jelly known from the fossil record, this exceptionally well preserved specimen provides important clues about the early evolution of the phylum Ctenophora and of metazoans in general. PMID- 17404243 TI - Identification of a regulator of transcription elongation as an accessory factor for the human Mediator coactivator. AB - The multiprotein Mediator coactivator complex is universally required for transcription of metazoan genes. It has been proposed to function by interfacing between transcriptional activators and the RNA polymerase II machinery. However, in vitro transcription systems reconstituted from homogeneous preparations of RNA polymerase II, the general transcription initiation factors, and the cofactor PC4 display relatively robust activator (HNF-4)-dependent activity, which, nonetheless, can be further stimulated by Mediator. By contrast, an unfractionated nuclear extract-based system in which Mediator has been immunodepleted displays a near-absolute dependence on ectopic Mediator. Here, we identified and purified an activity, MSA-2, that confers extract-like Mediator responsiveness to our reconstituted system. Mass spectrometric analyses identified its two constituent polypeptides as hSpt5 and hSpt4, which also comprise the elongation factor DSIF. Mechanistically, MSA-2/DSIF acts by restricting overall transcription in the pure system, thereby imposing a strong Mediator dependence. Our data thus point to potential mechanisms for Mediator function beyond its presently believed role in promoting the initial formation of the RNA polymerase II-containing preinitiation complex. PMID- 17404244 TI - Comment on "Cutting edge: induction of B7-H4 on APCs through IL-10: novel suppressive mode for regulatory T cells". PMID- 17404245 TI - AAIR: antibody antigen information resource. PMID- 17404246 TI - Comment on "A novel role of hypoxia-inducible factor in cobalt chloride- and hypoxia-mediated expression of IL-8 chemokine in human endothelial cells". PMID- 17404247 TI - Comment on "Aberrant regulation of synovial T cell activation by soluble costimulatory molecules in rheumatoid arthritis". PMID- 17404248 TI - Drosophila hemopoiesis and cellular immunity. AB - In Drosophila melanogaster larvae, three classes of circulating cellular immune surveillance cells (hemocytes) can be identified: plasmatocytes, crystal cells, and lamellocytes. Plasmatocytes are professional phagocytes most similar to the mammalian monocyte/macrophage lineage and make up approximately 95% of circulating hemocytes. The other approximately 5% of circulating hemocytes consists of crystal cells, which secrete components necessary for the melanization of invading organisms, as well as for wound repair. A third cell type known as lamellocytes are rarely seen in healthy larvae and are involved in the encapsulation of invading pathogens. There are no obvious mammalian counterparts for crystal cells or lamellocytes, and there is no equivalent to the lymphoid lineage in insects. In this review, I will discuss what is currently known about Drosophila hemopoiesis and the cellular immune response and where possible compare it to vertebrate mechanisms. PMID- 17404249 TI - Cutting edge: Tlr5-/- mice are more susceptible to Escherichia coli urinary tract infection. AB - Although TLR5 regulates the innate immune response to bacterial flagellin, it is unclear whether its function is essential during in vivo murine infections. To examine this question, we challenged Tlr5(-/-) mice transurethrally with Escherichia coli. At 2 days postinfection, wild-type mice exhibited increased inflammation of the bladder in comparison to Tlr5(-/-) mice. By day 5 postinfection, Tlr5(-/-) mice had significantly more bacteria in the bladders and kidneys in comparison to wild-type mice and showed increased inflammation in both organs. In addition, flagellin induced high levels of cytokine and chemokine expression in the bladder that was dependent on TLR5. Together, these data represent the first evidence that TLR5 regulates the innate immune response in the urinary tract and is essential for an effective murine in vivo immune response to an extracellular pathogen. PMID- 17404250 TI - Cutting edge: Antigen is not required for the activation and maintenance of virus specific memory CD8+ T cells in the lung airways. AB - Respiratory virus infections establish a population of memory CD8(+) T cells in the lung airways that persist for months after infection. However, the relationship between Ag-specific memory T cells in the lung airways and the systemic memory T cell pool is not well understood. The majority of lung airway memory T cells express a highly activated phenotype (CD69(+)/CD127(-)), suggesting that recent Ag stimulation is required to drive T cell activation and recruitment to the lung airways. In this study, we demonstrate that the lung airway environment itself in the absence of cognate Ag alters the expression of acute activation markers such as CD69 and CD127 on memory CD8(+) T cells. Furthermore, the steady-state recruitment of virus-specific memory CD8(+) T cells to the lung airways from the circulation can occur without recent Ag stimulation. These findings alter the current perceptions concerning the contribution of Ag to the maintenance of peripheral T cell memory. PMID- 17404251 TI - Cutting Edge: B cell receptor (BCR) cross-talk: the IL-4-induced alternate pathway for BCR signaling operates in parallel with the classical pathway, is sensitive to Rottlerin, and depends on Lyn. AB - B cell exposure to IL-4 alters subsequent BCR signaling such that ERK phosphorylation becomes signalosome-independent; however, the nature of this new, alternate signaling pathway and its relationship to the classical, signalosome dependent signaling pathway are not known. In this study, we report that the alternate and classical pathways for BCR signaling are differentially affected by rottlerin, and by Go6976 or LY294002, respectively. Furthermore, in B cells lacking protein kinase C (PKC)beta, the classical pathway for BCR signaling is blocked, whereas the alternate pathway is little affected. Conversely, in B cells lacking Lyn, the alternate pathway for BCR signaling is blocked, whereas the classical pathway is little affected. The rottlerin-sensitive element is not PKCdelta, inasmuch as the alternate pathway is not blocked in PKCdelta-deficient B cells. These results indicate that the rottlerin-sensitive, Lyn-dependent alternate pathway, and the classical pathway, for BCR signaling operate in parallel when BCR engagement follows IL-4 exposure. PMID- 17404253 TI - IL-7 Is essential for the development and the persistence of chronic colitis. AB - Although IL-7 has recently emerged as a key cytokine involved in controlling the homeostatic turnover and the survival of peripheral resting memory CD4(+) T cells, its potential to be sustained pathogenic CD4(+) T cells in chronic immune diseases, such as inflammatory bowel diseases, still remains unclear. In this study, we demonstrate that IL-7 is essential for the development and the persistence of chronic colitis induced by adoptive transfer of normal CD4(+)CD45RB(high) T cells or colitogenic lamina propria (LP) CD4(+) memory T cells into immunodeficient IL-7(+/+) x RAG-1(-/-) and IL-7(-/-) x RAG-1(-/-) mice. Although IL-7(+/+) x RAG-1(-/-) recipients transferred with CD4(+)CD45RB(high) splenocytes developed massive inflammation of the large intestinal mucosa concurrent with massive expansion of Th1 cells, IL-7(-/-) x RAG 1(-/-) recipients did not. Furthermore, IL-7(-/-) x RAG-1(-/-), but not IL-7(+/+) x RAG-1(-/-), mice transferred with LP CD4(+)CD44(high)CD62L(-)IL-7Ralpha(high) effector-memory T cells (T(EM)) isolated from colitic CD4(+)CD45RB(high) transferred mice did not develop colitis. Although rapid proliferation of transferred colitogenic LP CD4(+) T(EM) cells was observed in the in IL-7(-/-) x RAG-1(-/-) mice to a similar extent of those in IL-7(+/+) x RAG-1(-/-) mice, Bcl 2 expression was significantly down-modulated in the transferred CD4(+) T cells in IL-7(-/-) x RAG-1(-/-) mice compared with those in IL-7(+/+) x RAG-1(-/-) mice. Taken together, IL-7 is essential for the development and the persistence of chronic colitis as a critical survival factor for colitogenic CD4(+) T(EM) cells, suggesting that therapeutic approaches targeting IL-7/IL-7R signaling pathway may be feasible in the treatment of inflammatory bowel diseases. PMID- 17404252 TI - Cutting edge: recombinant Listeria monocytogenes expressing a single immune dominant peptide confers protective immunity to herpes simplex virus-1 infection. AB - The vast majority of the world's population is infected with HSV. Although antiviral therapy can reduce the incidence of reactivation and asymptomatic viral shedding, and limit morbidity and mortality from active disease, it cannot cure infection. Therefore, the development of an effective vaccine is an important global health priority. In this study, we demonstrate that recombinant Listeria monocytogenes (Lm) expressing the H-2K(b) glycoprotein B (gB)(498-505) peptide from HSV-1 triggers a robust CD8 T cell response to this Ag resulting in protective immunity to HSV infection. Following challenge with HSV-1, immune competent mice primed with recombinant Lm-expressing gB(498-505) Ag were protected from HSV-induced paralysis. Protection was associated with dramatic reductions in recoverable virus, and early expansion of HSV-1-specific CD8 T cells in the regional lymph nodes. Thus, recombinant Lm-expressing Ag from HSV represents a promising new class of vaccines against HSV infection. PMID- 17404254 TI - Dissociation of experimental allergic encephalomyelitis protective effect and allergic side reactions in tolerization with neuroantigen. AB - Administration of autoantigens under conditions that induce type 2 immunity frequently leads to protection from T cell-mediated autoimmune diseases. Such treatments, however, are inherently linked to the induction of IgG1 Abs and to the risk of triggering anaphylactic reactions. We studied the therapeutic benefit vs risk of immune deviation in experimental allergic encephalomyelitis of SJL mice induced by MP4, a myelin basic protein-proteolipid protein (PLP) fusion protein. MP4 administration in IFA induced type 2 T cell immunity, IgG1 Abs, and experimental allergic encephalomyelitis protection, and all three were enhanced by repeat injections. Despite high Ab titers, anaphylactic side reactions were not observed when MP4 was repeatedly injected in IFA or as soluble Ag s.c. In contrast, lethal anaphylaxis was seen after s.c. injection of soluble PLP:139-151 peptide, but not when the peptide was reinjected in IFA. Therefore, the Ab response accompanying the immune therapy constituted an anaphylactic risk factor only when the autoantigen was not retained in an adjuvant and when it was small enough to be readily disseminated within the body. Taken together, our data show that treatment regimens can be designed to boost the protective type 2 T cell response while avoiding the risk of Ab-mediated allergic side effects. PMID- 17404255 TI - Regulation of oxidative stress responses by ataxia-telangiectasia mutated is required for T cell proliferation. AB - Mutations in the gene encoding ataxia-telangiectasia (A-T) mutated (Atm) cause the disease A-T, characterized by immunodeficiency, the molecular basis of which is not known. Following stimulation through the TCR, Atm-deficient T cells and normal T cells in which Atm is inhibited undergo apoptosis rather than proliferation. Apoptosis is prevented by scavenging reactive oxygen species (ROS) during activation. Atm therefore plays a critical role in T cell proliferation by regulating responses to ROS generated following T cell activation. The inability of Atm-deficient T cells to control responses to ROS is therefore the molecular basis of immunodeficiency associated with A-T. PMID- 17404256 TI - NK cell maturation and peripheral homeostasis is associated with KLRG1 up regulation. AB - NK cells are important for the clearance of tumors, parasites, and virus-infected cells. Thus, factors that control NK cell numbers and function are critical for the innate immune response. A subset of NK cells express the inhibitory killer cell lectin-like receptor G1 (KLRG1). In this study, we identify that KLRG1 expression is acquired during periods of NK cell division such as development and homeostatic proliferation. KLRG1(+) NK cells are mature in phenotype, and we show for the first time that these cells have a slower in vivo turnover rate, reduced proliferative response to IL-15, and poorer homeostatic expansion potential compared with mature NK cells lacking KLRG1. Transfer into lymphopenic recipients indicate that KLRG1(-) NK cells are precursors of KLRG1(+) NK cells and KLRG1 expression accumulates following cell division. Furthermore, KLRG1(+) NK cells represent a significantly greater proportion of NK cells in mice with enhanced NK cell numbers such as Cd45(-/-) mice. These data indicate that NK cells acquire KLRG1 on their surface during development, and this expression correlates with functional distinctions from other peripheral NK cells in vivo. PMID- 17404257 TI - An essential role for IL-18 in CD8 T cell-mediated suppression of IgE responses. AB - The ability of CD8 T cells to suppress IgE responses is well established. Previously, we demonstrated that CD8 T cells inhibit IgE responses via the induction of IL-12, which promotes Th1 and suppresses Th2 responses. In this study, we show that IL-18 also plays an essential role in IgE suppression. In vitro, IL-18 synergized with IL-12 to promote Th1/T cytotoxic 1 and inhibit Th2/T cytotoxic 2 differentiation. OVA-specific TCR transgenic (OT-I) CD8 cells induced both IL-12 and IL-18 when cultured with OVA(257-264) peptide-pulsed dendritic cells. In vivo, IL-18(-/-) mice exhibited higher IgE and IgG1 levels compared with wild-type mice after immunization with OVA/alum. Furthermore, adoptive transfer of CD8 T cells from OVA-primed mice suppressed IgE responses in OVA/alum immunized mice, but not in IL-18(-/-) mice. IgE suppression in IL-18(-/-) mice was restored if CD8 T cells were coadoptively transferred with IL-18-competent wild-type bone marrow dendritic cell progenitors, demonstrating an essential role of IL-18 in CD8 T cell-mediated suppression of IgE responses. The data suggest that CD8 T cells induce IL-18 production during a cognate interaction with APCs that synergizes with IL-12 to promote immune deviation away from the allergic phenotype. Our data identify IL-18 induction as a potentially useful target in immunotherapy of allergic disease. PMID- 17404258 TI - IL-10 induces IL-10 in primary human monocyte-derived macrophages via the transcription factor Stat3. AB - IL-10 is an important immunosuppressive cytokine that can down-regulate expression of other cytokines and has been shown to down-regulate itself. We show, in this study, that treatment of human monocyte-derived macrophages with IL 10 induces IL-10 mRNA in a dose- and time-dependent manner with an optimum induction at 100 ng/ml and at 6 h, whereas IL-10-induced IL-10 protein can be detected at 18 h. In the same cells, IL-10 can partially suppress IL-10 mRNA induced by LPS, but only down to the level of IL-10-induced IL-10. An adenoviral luciferase reporter construct driven by the -195 IL-10 promoter, which contains a Stat motif, was readily induced by both IL-10 and LPS. Mutation of this Stat motif ablated IL-10 activation of this promoter, but not the LPS activation. Finally, we show that overexpression of a dominant-negative Stat3 protein will prevent IL-10 induction, but not LPS induction, of IL-10 mRNA. These data show that IL-10 induces IL-10 in monocyte-derived macrophages in an autocrine manner via activation of the transcription factor Stat3. PMID- 17404259 TI - Profound enhancement of the IL-12/IL-18 pathway of IFN-gamma secretion in human CD8+ memory T cell subsets via IL-15. AB - Human memory CD8(+) T cell subsets, termed central memory and effector memory T cells, can be identified by expression of CD45RA, CD62 ligand (CD62L), and CCR7. Accordingly, functional differences have been described for each subset, reflecting unique roles in immunological memory. The common gamma-chain cytokines IL-15 and IL-7 have been shown to induce proliferation and differentiation of human CD8(+) T cell subsets, as well as increased effector functions (i.e., cytokines, cytotoxicity). In this study, we observed that addition of IL-15 or IL 7 to cultures of human CD8(+) T cells profoundly enhanced the IL-12-IL-18 pathway of IFN-gamma production. Importantly, IL-15 and IL-7 lowered the threshold concentrations of IL-12 and IL-18 required for induction of IFN-gamma by 100 fold. Comparison of IL-15 and IL-7 demonstrated that IL-15 was superior in its ability to enhance IL-12-IL-18-induced IFN-gamma, without evidence of a synergistic effect between IL-15 and IL-7. We also observed that IL-15- and IL-7 mediated enhancement of IL-12-IL-18-induced IFN-gamma production was a functional property of effector memory CD8(+) T cells. Despite a lack of association between cell division and acquisition of IL-12-IL-18-induced IFN-gamma, down-regulation of CD62L expression correlated well with increased IL-12-IL-18-induced IFN-gamma. Purified central memory T cells stimulated with IL-15 and IL-7 down-regulated CD62L and acquired potent IL-12-IL-18-induced IFN-gamma similar to effector memory T cells. Thus, in addition to its known role in development of T cell memory, IL-15 may amplify memory CD8(+) T cell effector functions by increasing sensitivity to proinflammatory cytokine stimulation. PMID- 17404260 TI - Spontaneous autoreactive memory B cell formation driven by a high frequency of autoreactive CD4+ T cells. AB - Although somatically mutated autoantibodies are characteristic of many autoimmune diseases, the processes that can lead to their development remain poorly understood. We have examined the formation of autoreactive memory B cells in PevHA mice, which express the influenza virus PR8 hemagglutinin (HA) as a transgenic membrane bound neo-self-Ag. Using a virus immunization strategy, we show that PR8 HA-specific memory B cell formation can occur in PevHA mice, even though a major subset of PR8 HA-specific B cells is negatively selected from the primary repertoire. Moreover, PR8 HA-specific memory B cells develop spontaneously in TS1 x PevHA mice, which coexpress a transgenic PR8 HA-specific TCR and contain a high frequency of HA-specific CD4(+) T cells. Notably, autoreactive memory B cell formation occurred in TS1 x PevHA mice even though approximately half of the HA-specific CD4(+) T cells were CD25(+)Foxp3(+) cells that could significantly attenuate, but did not completely abolish HA-specific autoantibody production in an adoptive transfer setting. The findings provide evidence that a high frequency of autoreactive CD4(+) T cells can be sufficient to promote autoreactive memory B cell formation in the absence of signals provided by overt immunization or infection and despite the presence of abundant autoantigen-specific CD4(+)CD25(+)Foxp3(+) regulatory T cells. PMID- 17404262 TI - The A subunit of type IIb enterotoxin (LT-IIb) suppresses the proinflammatory potential of the B subunit and its ability to recruit and interact with TLR2. AB - The type IIb heat-labile enterotoxin of Escherichia coli (LT-IIb) and its nontoxic pentameric B subunit (LT-IIb-B(5)) display different immunomodulatory activities, the mechanisms of which are poorly understood. We investigated mechanisms whereby the absence of the catalytically active A subunit from LT-IIb B(5) renders this molecule immunostimulatory through TLR2. LT-IIb-B(5), but not LT-IIb, induced TLR2-mediated NF-kappaB activation and TNF-alpha production. These LT-IIb-B(5) activities were antagonized by LT-IIb; however, inhibitors of adenylate cyclase or protein kinase A reversed this antagonism. The LT-IIb antagonistic effect is thus likely dependent upon the catalytic activity of its A subunit, which causes elevation of intracellular cAMP and activates cAMP dependent protein kinase A. Consistent with this, a membrane-permeable cAMP analog and a cAMP-elevating agonist, but not catalytically defective point mutants of LT-IIb, mimicked the antagonistic action of wild-type LT-IIb. The mutants moreover displayed increased proinflammatory activity compared with wild type LT-IIb. Additional mechanisms for the divergent effects on TLR2 activation by LT-IIb and LT-IIb-B(5) were suggested by findings that the latter was significantly stronger in inducing lipid raft recruitment of TLR2 and interacting with this receptor. The selective use of TLR2 by LT-IIb-B(5) was confirmed in an assay for IL-10, which is inducible by both LT-IIb and LT-IIb-B(5) at comparable levels; TLR2-deficient macrophages failed to induce IL-10 in response to LT-IIb B(5) but not in response to LT-IIb. These differential immunomodulatory effects by LT-IIb and LT-IIb-B(5) have important implications for adjuvant development and, furthermore, suggest that enterotoxic E. coli may suppress TLR-mediated innate immunity through the action of the enterotoxin A subunit. PMID- 17404261 TI - Dendritic cells from lupus-prone mice are defective in repressing immunoglobulin secretion. AB - Autoimmunity results from a breakdown in tolerance mechanisms that regulate autoreactive lymphocytes. We recently showed that during innate immune responses, secretion of IL-6 by dendritic cells (DCs) maintained autoreactive B cells in an unresponsive state. In this study, we describe that TLR4-activated DCs from lupus prone mice are defective in repressing autoantibody secretion, coincident with diminished IL-6 secretion. Reduced secretion of IL-6 by MRL/lpr DCs reflected diminished synthesis and failure to sustain IL-6 mRNA production. This occurred coincident with lack of NF-kappaB and AP-1 DNA binding and failure to sustain IkappaBalpha phosphorylation. Analysis of individual mice showed that some animals partially repressed Ig secretion despite reduced levels of IL-6. This suggests that in addition to IL-6, DCs secrete other soluble factor(s) that regulate autoreactive B cells. Collectively, the data show that MRL/lpr mice are defective in DC/IL-6-mediated tolerance, but that some individuals maintain the ability to repress autoantibody secretion by an alternative mechanism. PMID- 17404263 TI - Differential expression and function of phosphodiesterase 4 (PDE4) subtypes in human primary CD4+ T cells: predominant role of PDE4D. AB - Type 4 phosphodiesterases (PDE4) are critical regulators in TCR signaling by attenuating the negative constraint of cAMP. In this study, we show that anti CD3/CD28 stimulation of human primary CD4(+) T cells increases the expression of the PDE4 subtypes PDE4A, PDE4B, and PDE4D in a specific and time-dependent manner. PDE4A and PDE4D mRNAs as well as enzyme activities were up-regulated within 5 days, PDE4B showed a transient up-regulation with highest levels after 24 h. The induction was shown to be independent of different stimulation conditions and was similar in naive and memory T cell subpopulations. To elucidate the functional impact of individual PDE4 subtypes on T cell function, we used PDE4 subtype-specific short-interfering RNAs (siRNAs). Knockdown of either PDE4B or PDE4D inhibited IL-2 release 24 h after stimulation (time point of maximal IL-2 concentrations) to an extent similar to that observed with the panPDE4 inhibitor RP73401 (piclamilast). Substantial amounts of IFN-gamma or IL-5 were measured only at later time points. siRNA targeting PDE4D showed a predominant inhibitory effect on these cytokines measured after 72 h. However, the inhibition of all cytokines was most effective when PDE4 siRNAs were applied in combination. Although the effect of PDE4 inhibition on T cell proliferation is small, the PDE4D-targeting siRNA alone was as effective as the panPDE4 inhibitor, whereas PDE4A or PDE4B siRNAs had hardly an effect. In summary, individual PDE4 subtypes have overall nonredundant, but complementary, time-dependent roles in propagating various T cell functions and PDE4D is the form likely playing a predominant role. PMID- 17404264 TI - IFN-alpha-induced signal transduction, gene expression, and antitumor activity of immune effector cells are negatively regulated by suppressor of cytokine signaling proteins. AB - Proteins belonging to the suppressors of cytokine signaling (SOCS) family have been shown to regulate cytokine signal transduction in various cell types but their role in modulating the response of immune cells to IFN-alpha has not been fully explored. We hypothesized that SOCS proteins would inhibit the antitumor activity of IFN-alpha-stimulated immune cells. Transcripts for SOCS1, SOCS2, SOCS3, and cytokine-inducible Src homology 2-containing protein were identified in total human PBMC (PBMCs, NK cells, and T cells) within 1-2 h of stimulation with IFN-alpha (10(3)-10(5) U/ml). Immunoblot analysis confirmed the expression of these factors at the protein level. Transcripts for SOCS proteins were rapidly but variably induced in PBMCs from patients with metastatic melanoma following the i.v. administration of IFN-alpha-2b (20 million units/m(2)). Overexpression of SOCS1 and SOCS3, but not SOCS2, in the Jurkat T cell line inhibited IFN-alpha induced phosphorylated STAT1 and the transcription of IFN-stimulated genes. Conversely, small inhibitory RNA-mediated down-regulation of SOCS1 and SOCS3 in Jurkat cells and normal T cells enhanced the transcriptional response to IFN alpha. Loss of SOCS1 or SOCS3 in murine immune effectors was associated with enhanced IFN-induced phosphorylated STAT1, transcription of IFN-stimulated genes, and antitumor activity. Of note, IFN-alpha treatment eliminated melanoma tumors in 70% of SOCS1-deficient mice, whereas IFN-treated SOCS-competent mice all died. The antitumor effects of IFN-alpha in tumor-bearing SOCS1-deficient mice were markedly inhibited following depletion of CD8(+) T cells. These results indicate that the antitumor response of immune effector cells to exogenous IFN-alpha is regulated by SOCS proteins. PMID- 17404265 TI - Glycoprotein 120 binding to CXCR4 causes p38-dependent primary T cell death that is facilitated by, but does not require cell-associated CD4. AB - HIV-1 infection causes the depletion of host CD4 T cells through direct and indirect (bystander) mechanisms. Although HIV Env has been implicated in apoptosis of uninfected CD4 T cells via gp120 binding to either CD4 and/or the chemokine receptor 4 (CXCR4), conflicting data exist concerning the molecular mechanisms involved. Using primary human CD4 T cells, we demonstrate that gp120 binding to CD4 T cells activates proapoptotic p38, but does not activate antiapoptotic Akt. Because ligation of the CD4 receptor alone or the CXCR4 receptor alone causes p38 activation and apoptosis, we used the soluble inhibitors, soluble CD4 (sCD4) or AMD3100, to delineate the role of CD4 and CXCR4 receptors, respectively, in gp120-induced p38 activation and death. sCD4 alone augments gp120-induced death, suggesting that CXCR4 signaling is principally responsible. Supporting that model, AMD3100 reduces death caused by gp120 or by gp120/sCD4. Finally, prevention of gp120-CXCR4 interaction with 12G5 Abs blocks p38 activation and apoptosis, whereas inhibition of CD4-gp120 interaction with Leu-3a has no effect. Consequently, we conclude that gp120 interaction with CXCR4 is required for gp120 apoptotic effects in primary human T cells. PMID- 17404266 TI - Aggregated ursolic acid, a natural triterpenoid, induces IL-1beta release from murine peritoneal macrophages: role of CD36. AB - IL-1beta has been shown to play a pivotal role in the development of inflammatory disorders. We recently found that a natural triterpene, ursolic acid (UA), enhanced MIF release from nonstimulated macrophages. In this study, we examined the effects of UA on the production of several cytokines in resident murine peritoneal macrophages (pMphi). UA increased the protein release of IL-1beta, IL 6, and MIF, but not of TNF-alpha, in dose- and time-dependent manners. This triterpene also strikingly induced the activation of p38 MAPK and ERK1/2 together with that of upstream kinases. The release of UA-induced IL-1beta was significantly inhibited by the inhibitors of p38 MAPK, MEK1/2, ATP-binding cassette transporter, and caspase-1. Furthermore, UA induced intracellular ROS generation for IL-1beta production, which was suppressed by an antioxidant. Pretreatment with an anti-CD36 Ab significantly suppressed IL-1beta release, and surface plasmon resonance assay results showed that UA bound to CD36 on macrophages. In addition, the amount of IL-1beta released from UA-treated pMphi of CD36-deficient mice was markedly lower than that from those of wild-type mice. Interestingly, UA was found to aggregate in culture medium, and the aggregates were suggested to be responsible for IL-1beta production. In addition, i.p. administration of UA increased the levels of IL-1beta secretion and MPO activity in colonic mucosa of ICR mice. Taken together, our results indicate that aggregated UA is recognized, in part, by CD36 on macrophages for generating ROS, thereby activating p38 MAPK, ERK1/2, and caspase-1, as well as releasing IL-1beta protein via the ATP-binding cassette transporter. PMID- 17404267 TI - T cell repertoire diversity is required for relapses in myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis. AB - Comparison of TCRalphabeta repertoires of myelin oligodendrocyte glycoprotein (MOG)-specific T lymphocytes in C57BL/6 and TdT-deficient littermates (TdT(-/-)) generated during experimental autoimmune encephalomyelitis (EAE) highlights a link between a diversified TCRalphabeta repertoire and EAE relapses. At the onset of the disease, the EAE-severity is identical in TdT(+/-) and TdT(-/-) mice and the neuropathologic public MOG-specific T cell repertoires express closely similar public Valpha-Jalpha and Vbeta-Jbeta rearrangements in both strains. However, whereas TdT(+/+) and TdT(+/-) mice undergo successive EAE relapses, TdT( /-) mice recover definitively and the lack of relapses does not stem from dominant regulatory mechanisms. During the first relapse of the disease in TdT(+/ ) mice, new public Valpha-Jalpha and Vbeta-Jbeta rearrangements emerge that are distinct from those detected at the onset of the disease. Most of these rearrangements contain N additions and are found in CNS-infiltrating T lymphocytes. Furthermore, CD4(+) T splenocytes bearing these rearrangements proliferate to the immunodominant epitope of MOG and not to other immunodominant epitopes of proteolipid protein and myelin basic protein autoantigens, excluding epitope spreading to these myelin proteins. Thus, in addition to epitope spreading, a novel mechanism involving TCRalphabeta repertoire diversification contributes to autoimmune progression. PMID- 17404268 TI - Beta1-adrenergic receptors on immune cells impair innate defenses against Listeria. AB - Cold restraint (CR) for 1 h elicits a psychological and physiological stress that inhibits host defenses against Listeria monocytogenes (LM). Previous analyses indicated that this inhibition is not due to depletion of B or T cells but is instead dependent on signaling through beta-adrenoceptors (betaARs). We now show that impaired host resistance by CR cannot be accounted for by a decrease in LM specific (listeriolysin O(91-99) tetramer(+)) effector CD8(+) T cells; this result is consistent with previous observations that CR-induced effects are mainly limited to early anti-LM responses. beta2-Adrenoceptor (beta2AR)(-/-) FVB/NJ and wild-type FVB/NJ mice had equivalent anti-LM defenses, whereas beta1 adrenoceptor (beta1AR)(-/-) FVB/NJ mice had lower levels of LM even when subjected to CR treatment. Additionally, host-resistance competency of beta1AR(-/ ) mice could be transferred to irradiated wild-type mice reconstituted with beta1AR(-/-) bone marrow progenitors and spleen cells, indicating that beta1AR signaling on immune cells reduces anti-LM responses. beta1AR(-/-) mice had improved cellular (delayed-type hypersensitivity) responses while beta2AR(-/-) mice had improved humoral responses (IgG1, IgG2, and IgM), a result that further explains the strain differences in LM defenses. CR-induced expression of beta1AR and beta2AR mRNA was assessed by real-time PCR. CR treatment significantly increased betaAR mRNAs in Ficoll-purified and F4/80(+)-enhanced liver but not splenic homogenates, demonstrating an organ-specific effect of stress that alters host defenses. Finally, CR treatment induced early increases in perforin expression that may enhance immune cell apoptosis and interfere with LM clearance. In conclusion, beta1AR signaling has immunomodulatory effects on early cell-mediated immune responses; a lack of beta1AR signaling improves antilisterial defenses and cell-mediated immunity, in general. PMID- 17404269 TI - Type 1 sphingosine 1-phosphate G protein-coupled receptor (S1P1) mediation of enhanced IL-4 generation by CD4 T cells from S1P1 transgenic mice. AB - Sphingosine 1-phosphate (S1P) is a natural lipid mediator that regulates immune cell traffic, Ab production, and T cell cytokine generation by mechanisms that enhance Th2 activities. Responses to S1P are controlled principally by the diverse expression patterns of its receptors in different cells. In T cells, the type 1 (S1P(1)) and type 4 (S1P(4)) G protein-coupled receptors are predominant. S1P(1) mainly transduces effects on T cell migration and trafficking, whereas S1P(4) transduces immunosuppression via its effects on T cell proliferation and cytokine production. Using T cell-specific S1P(1) transgenic (TG) mice, we investigated the regulatory effects of the S1P-S1P(1) axis on T cell cytokine production. The production of IL-4, but not IL-2 or IFN-gamma, was significantly up-regulated >10-fold in activated CD4 T cells from S1P(1) TG mice compared with those from wild-type mice. Quantitative real-time PCR analysis revealed that IL-4 up-regulation was initiated at the mRNA level as early as 4 h after T cell activation. The up-regulation of IL-4 mRNA was mediated by c-Maf, Jun B, and Gata3 as demonstrated by increases in their protein expression and DNA-binding activities. In contrast, the expression and DNA-binding activities of T-bet, FosB, C-Fos, Jun D, Fra-1, Fra-2, and c-Jun all were identical in wild-type and TG CD4 T cells. Immunological assays showed that increased IL-4 levels induced greater production of IgE. Thus, the S1P-S1P(1) axis specifically up-regulates c Maf, Jun B, and Gata3, which consequently enhance IL-4 production that may lead to a Th2 phenotype. PMID- 17404270 TI - Human peripheral blood T regulatory cells (Tregs), functionally primed CCR4+ Tregs and unprimed CCR4- Tregs, regulate effector T cells using FasL. AB - Regulatory CD25(+)CD4(+) T cells (Tregs) play an important role in the control of peripheral tolerance. In this study we demonstrate that human peripheral blood Tregs can be divided into two distinct populations based on the expression of CCR4. The majority ( approximately 75%) of freshly isolated Tregs express CCR4 and presumably represent memory-type Tregs. Interestingly, CCR4(-) Tregs require anti-CD3 Ab-mediated activation to acquire a regulatory activity, while CCR4(+) Tregs appear to be already primed to suppress the proliferation of CD8(+) T cells. CCR4 is also expressed on CD25(low)CD4(+) T cells (CCR4(+) non-Tregs) that mostly suppress Th1-type polarization without affecting T cell proliferation, presumably via the production of immunomodulatory cytokines like IL-10. In contrast, CCR4(+) Tregs express FasL to primarily regulate T cell proliferation via a contact-mediated process involving FasL/Fas signaling, a major regulatory pathway of T cell homeostasis. Finally, we also demonstrate that the depletion of CCR4(+) T cells leads to Th1-type polarization of CD4(+) T cells and augmentation of CD8(+) T cell responses to tumor Ags. PMID- 17404271 TI - Stat3 and Stat4 direct development of IL-17-secreting Th cells. AB - IL-17-secreting CD4(+) T cells are critically involved in inflammatory immune responses. Development of these cells is promoted in vivo and in vitro by IL-23 or TGFbeta1 plus IL-6. Despite growing interest in this inflammatory Th subset, little is known about the transcription factors that are required for their development. We demonstrate that Stat3 is required for programming the TGFbeta1 plus IL-6 and IL-23-stimulated IL-17-secreting phenotype, as well as for RORgammat expression in TGFbeta1 plus IL-6-primed cells. Moreover, retroviral transduction of a constitutively active Stat3 into differentiating T cell cultures enhances IL-17 production from these cells. We further show that Stat4 is partially required for the development of IL-23-, but not TGFbeta1 plus IL-6 primed IL-17-secreting cells, and is absolutely required for IL-17 production in response to IL-23 plus IL-18. The requirements for Stat3 and Stat4 in the development of these IL-17-secreting subsets reveal additional mechanisms in Th cell fate decisions during the generation of proinflammatory cell types. PMID- 17404272 TI - Interdependency of MHC class II/self-peptide and CD1d/self-glycolipid presentation by TNF-matured dendritic cells for protection from autoimmunity. AB - Dendritic cells (DC) are key regulators of T cell immunity and tolerance. NKT cells are well-known enhancers of Th differentiation and regulatory T cell function. However, the nature of the DC directing T and NKT cell activation and polarization as well as the role of the respective CD1d Ags presented is still unclear. In this study, we show that peptide-specific CD4(+)IL-10(+) T cell mediated full experimental autoimmune encephalomyelitis (EAE) protection by TNF treated semimatured DCs was dependent on NKT cells recognizing an endogenous CD1d ligand. NKT cell activation by TNF-matured DCs induced high serum levels of IL-4 and IL-13 which are absent in NKT cell-deficient mice, whereas LPS plus anti-CD40 treated fully mature DCs induce serum IFN-gamma. In the absence of IL-4Ralpha chain signaling or NKT cells, no complete EAE protection was achieved by TNF-DCs, whereas transfer of NKT cells into Jalpha281(-/-) mice restored it. However, activation of NKT cells alone was not sufficient for EAE protection and early serum Th2 deviation. Simultaneous activation of NKT cells and CD4(+) T cells by the same DC was required for EAE protection. Blocking experiments demonstrated that NKT cells recognize an endogenous glycolipid presented on CD1d on the injected DC. Together, this indicates that concomitant and interdependent presentation of MHC II/self-peptide and CD1d/self-isoglobotrihexosylceramide to T and NKT cells by the same partially or fully matured DC determines protective and nonprotective immune responses in EAE. PMID- 17404273 TI - TLR2-mediated survival of Staphylococcus aureus in macrophages: a novel bacterial strategy against host innate immunity. AB - TLR2 plays a role as a pattern-recognition receptor in the innate immune response involving secreted proteins against microbial pathogens. To examine its possible involvement in the cellular response, we determined the levels of the engulfment and subsequent killing of bacteria by macrophages prepared from TLR2-deficient and wild-type mice. The level of the engulfment of Staphylococcus aureus or Escherichia coli was almost the same between TLR2-lacking and wild-type macrophages. However, the colony-forming ability of engulfed S. aureus, but not of E. coli, decreased to a greater extent in TLR2-lacking macrophages than in the wild-type control. The incubation with S. aureus caused activation of JNK in wild type macrophages but not in TLR2-lacking macrophages, and the pretreatment of wild-type macrophages with a JNK inhibitor increased the rate of killing of engulfed S. aureus, but again not of E. coli. In addition, the number of colonies formed by engulfed S. aureus increased in the JNK-dependent manner when TLR2 lacking macrophages were pretreated with LPS. Furthermore, JNK seemed to inhibit the generation of superoxide, not of NO, in macrophages. These results collectively suggested that the level of superoxide is reduced in macrophages that have engulfed S. aureus through the actions of TLR2-activated JNK, resulting in the prolonged survival of the bacterium in phagosomes. The same regulation did not influence the survival of E. coli, because this bacterium was more resistant to superoxide than S. aureus. We propose a novel bacterial strategy for survival in macrophages involving the hijacking of an innate immune receptor. PMID- 17404274 TI - Schnurri-2 controls memory Th1 and Th2 cell numbers in vivo. AB - Schnurri-2 (Shn-2) is a large zinc-finger containing protein, and it plays a critical role in cell growth, signal transduction and lymphocyte development. In Shn-2-deficient CD4 T cells, the activation of NF-kappaB was up-regulated and their ability to differentiate into Th2 cells was enhanced. We herein demonstrate that Th1 and Th2 memory cells are not properly generated from Shn-2-deficient effector Th1/Th2 cells. Even a week after the transfer of effector Th1/Th2 cells into syngeneic mice, a dramatic decrease in the number of Shn-2-deficient donor T cells was detected particularly in the lymphoid organs. The transferred Shn-2 deficient Th1/Th2 cells express higher levels of the activation marker CD69. No significant defect in the BrdU incorporation in the Shn-2-deficient transferred CD4 T cells was observed. The numbers of apoptotic cells were selectively higher in Shn-2-deficient donor Th1/Th2 cell population. Moreover, Shn-2-deficient effector Th1 and Th2 cells showed an increased susceptibility to cell death in in vitro cultures with increased expression of FasL. Transfer of Th2 effector cells over-expressing the p65 subunit of NF-kappaB resulted in a decreased number of p65-expressing cells in the lymphoid organs. As expected, T cell-dependent Ab responses after in vivo immunization of Shn-2-deficient mice were significantly reduced. Thus, Shn-2 appears to control the generation of memory Th1/Th2 cells through a change in their susceptibility to cell death. PMID- 17404275 TI - Intestinal lamina propria retaining CD4+CD25+ regulatory T cells is a suppressive site of intestinal inflammation. AB - It is well known that immune responses in the intestine remain in a state of controlled inflammation, suggesting that not only does active suppression by regulatory T (T(REG)) cells play an important role in the normal intestinal homeostasis, but also that its dysregulation of immune response leads to the development of inflammatory bowel disease. In this study, we demonstrate that murine CD4(+)CD25(+) T cells residing in the intestinal lamina propria (LP) constitutively express CTLA-4, glucocorticoid-induced TNFR, and Foxp3 and suppress proliferation of responder CD4(+) T cells in vitro. Furthermore, cotransfer of intestinal LP CD4(+)CD25(+) T cells prevents the development of chronic colitis induced by adoptive transfer of CD4(+)CD45RB(high) T cells into SCID mice. When lymphotoxin (LT)alpha-deficient intercrossed Rag2 double knockout mice (LTalpha(-/-) x Rag2(-/-)), which lack mesenteric lymph nodes and Peyer's patches, are transferred with CD4(+)CD45RB(high) T cells, they develop severe wasting disease and chronic colitis despite the delayed kinetics as compared with the control LTalpha(+/+) x Rag2(-/-) mice transferred with CD4(+)CD45RB(high) T cells. Of note, when a mixture of splenic CD4(+)CD25(+) T(REG) cells and CD4(+)CD45RB(high) T cells are transferred into LTalpha(-/-) x Rag2(-/-) recipients, CD4(+)CD25(+) T(REG) cells migrate into the colon and prevent the development of colitis in LTalpha(-/-) x Rag2(-/-) recipients as well as in the control LTalpha(+/+) x Rag2(-/-) recipients. These results suggest that the intestinal LP harboring CD4(+)CD25(+) T(REG) cells contributes to the intestinal immune suppression. PMID- 17404276 TI - CD56brightCD16- killer Ig-like receptor- NK cells display longer telomeres and acquire features of CD56dim NK cells upon activation. AB - Human NK cells can be divided into CD56(dim)CD16(+) killer Ig-like receptors (KIR)(+/-) and CD56(bright)CD16(-) KIR(-) subsets that have been characterized extensively regarding their different functions, phenotype, and tissue localization. Nonetheless, the developmental relationship between these two NK cell subsets remains controversial. We report that, upon cytokine activation, peripheral blood (PB)-CD56(bright) NK cells mainly gain the signature of CD56(dim) NK cells. Remarkably, KIR can be induced not only on CD56(bright), but also on CD56(dim) KIR(-) NK cells, and their expression correlates with lower proliferative response. In addition, we demonstrate for the first time that PB CD56(dim) display shorter telomeres than PB- and lymph node (LN)-derived CD56(bright) NK cells. Along this line, although human NK cells collected from nonreactive LN display almost no KIR and CD16 expression, NK cells derived from highly reactive LN, efferent lymph, and PB express significant amounts of KIR and CD16, implying that CD56(bright) NK cells could acquire these molecules in the LN during inflammation and then circulate through the efferent lymph into PB as KIR(+)CD16(+) NK cells. Altogether, our results suggest that CD56(bright)CD16(-) KIR(-) and CD56(dim)CD16(+)KIR(+/-) NK cells correspond to sequential steps of differentiation and support the hypothesis that secondary lymphoid organs can be sites of NK cell final maturation and self-tolerance acquisition during immune reaction. PMID- 17404277 TI - A unique thymic fibroblast population revealed by the monoclonal antibody MTS-15. AB - T cell differentiation in the thymus is dependent upon signals from thymic stromal cells. Most studies into the nature of these signals have focused only on the support provided by the thymic epithelium, but there is an emerging view that other stromal cells such as mesenchymal fibroblasts may also be involved. Study of the latter has been hindered by a lack of appropriate markers, particularly those allowing their isolation. In this study, we describe a new surface marker of thymic stroma, MTS-15, and demonstrate its specificity for fibroblasts and a subset of endothelial cells. Coculture experiments showed that the determinant could be transferred between cells. Extensive biochemical analysis demonstrated that the Ag bound by MTS-15 was the glycosphingolipid Forssman determinant, consistent with the distribution observed. Transcriptional analysis of purified MTS-15(+) thymic fibroblasts revealed a unique expression profile for a number of chemokines and growth factors important to thymocyte and epithelial cell development. In a model of cyclophosphamide-induced thymic involution and regeneration, fibroblasts were found to expand extensively and express growth factors important to epithelial proliferation and increased T cell production just before thymic regeneration. Overall, this study identifies a useful marker of thymic fibroblasts and highlights this subpopulation as a key player in thymic function by virtue of their support of both thymocytes and epithelial cells. PMID- 17404278 TI - Dermal fibroblasts induce maturation of dendritic cells. AB - To trigger an effective T cell-mediated immune response in the skin, cutaneous dendritic cells (DC) migrate into locally draining lymph nodes, where they present Ag to naive T cells. Little is known about the interaction of DC with the various cellular microenvironments they encounter during their migration from the skin to lymphoid tissues. In this study, we show that human DC generated from peripheral blood monocytes specifically interact with human dermal fibroblasts via the interaction of beta(2) integrins on DC with Thy-1 (CD90) and ICAM-1 on fibroblasts. This induced the phenotypic maturation of DC reflected by expression of CD83, CD86, CD80, and HLA-DR in a TNF-alpha- and ICAM-1-dependent manner. Moreover, fibroblast-matured DC potently induced T cell activation reflected by CD25 expression and enhanced T cell proliferation. Together these data demonstrate that dermal fibroblasts that DC can encounter during their trafficking from skin to lymph node can act as potent regulators of DC differentiation and function, and thus may actively participate in the regulation and outcome of DC-driven cutaneous immune responses. PMID- 17404279 TI - Mannose receptor expression and function define a new population of murine dendritic cells. AB - In vitro the mannose receptor (MR) mediates Ag internalization by dendritic cells (DC) and favors the presentation of mannosylated ligands to T cells. However, in vivo MR seems to play a role not in Ag presentation but in the homeostatic clearance of endogenous ligands, which could have the secondary benefit of reducing the levels of endogenous Ag available for presentation to the adaptive immune system. We have now observed that while MR(+) cells are consistently absent from T cell areas of spleen and mesenteric lymph nodes (LN), peripheral LN of untreated adult mice contain a minor population of MR(+)MHCII(+) in the paracortex. This novel MR(+) cell population can be readily identified by flow cytometry and express markers characteristic of DC. Furthermore, these MR(+) DC like cells located in T cell areas can be targeted with MR ligands (anti-MR mAb). Numbers of MR(+)MHCII(+) cells in the paracortex are increased upon stimulation of the innate immune system and, accordingly, the amount of anti-MR mAb reaching MR(+)MHCII(+) cells in T cell areas is dramatically enhanced under these conditions. Our results indicate that the MR can act as an Ag-acquisition system in a DC subpopulation restricted to lymphoid organs draining the periphery. Moreover, the effect of TLR agonists on the numbers of these MR(+) DC suggests that the immunogenicity of MR ligands could be under the control of innate stimulation. In accordance with these observations, ligands highly specific for the MR elicit enhanced humoral responses in vivo only when administered in combination with endotoxin. PMID- 17404280 TI - Quantitative network signal combinations downstream of TCR activation can predict IL-2 production response. AB - Proximal signaling events activated by TCR-peptide/MHC (TCR-pMHC) binding have been the focus of intense ongoing study, but understanding how the consequent downstream signaling networks integrate to govern ultimate avidity-appropriate TCR-pMHC T cell responses remains a crucial next challenge. We hypothesized that a quantitative combination of key downstream network signals across multiple pathways must encode the information generated by TCR activation, providing the basis for a quantitative model capable of interpreting and predicting T cell functional responses. To this end, we measured 11 protein nodes across six downstream pathways, along five time points from 10 min to 4 h, in a 1B6 T cell hybridoma stimulated by a set of three myelin proteolipid protein 139-151 altered peptide ligands. A multivariate regression model generated from this data compendium successfully comprehends the various IL-2 production responses and moreover successfully predicts a priori the response to an additional peptide treatment, demonstrating that TCR binding information is quantitatively encoded in the downstream network. Individual node and/or time point measurements less effectively accounted for the IL-2 responses, indicating that signals must be integrated dynamically across multiple pathways to adequately represent the encoded TCR signaling information. Of further importance, the model also successfully predicted a priori direct experimental tests of the effects of individual and combined inhibitors of the MEK/ERK and PI3K/Akt pathways on this T cell response. Together, our findings show how multipathway network signals downstream of TCR activation quantitatively integrate to translate pMHC stimuli into functional cell responses. PMID- 17404281 TI - Increased hyperoxia-induced mortality and acute lung injury in IL-13 null mice. AB - IL-13 is a critical effector at sites of Th2 inflammation and remodeling. As a result, anti-IL-13-based therapies are being actively developed to treat a variety of diseases and disorders. However, the beneficial effects of endogenous IL-13 in the normal and diseased lung have not been adequately defined. We hypothesized that endogenous IL-13 is an important regulator of oxidant-induced lung injury and inflammation. To test this hypothesis, we compared the effects of 100% O(2) in mice with wild-type and null IL-13 loci. In this study, we demonstrate that hyperoxia significantly augments the expression of the components of the IL-13R, IL-13Ralpha1, and IL-4Ralpha. We also demonstrate that, in the absence of IL-13, hyperoxia-induced tissue inflammation is decreased. In contrast, in the IL-13 null mice, DNA injury, cell death, caspase expression, and activation and mortality are augmented. Interestingly, the levels of the cytoprotective cytokines vascular endothelial cell growth factor, IL-6, and IL-11 were decreased in the bronchoalveolar lavage fluid. These studies demonstrate that the expression of the IL-13R is augmented and that the endogenous IL-13-IL 13R pathway contributes to the induction of inflammation and the inhibition of injury in hyperoxic acute lung injury. PMID- 17404282 TI - Protective role of macrophages in noninflammatory lung injury caused by selective ablation of alveolar epithelial type II Cells. AB - Macrophages have a wide variety of activities and it is largely unknown how the diverse phenotypes of macrophages contribute to pathological conditions in the different types of tissue injury in vivo. In this study we established a novel animal model of acute respiratory distress syndrome caused by the dysfunction of alveolar epithelial type II (AE2) cells and examined the roles of alveolar macrophages in the acute lung injury. The human diphtheria toxin (DT) receptor (DTR), heparin-binding epidermal growth factor-like growth factor (HB-EGF), was expressed under the control of the lysozyme M (LysM) gene promoter in the mice. When DT was administrated to the mice they suffered from acute lung injury and died within 4 days. Immunohistochemical examination revealed that AE2 cells as well as alveolar macrophages were deleted via apoptosis in the mice treated with DT. Consistent with the deletion of AE2 cells, the amount of surfactant proteins in bronchoalveolar lavage fluid was greatly reduced in the DT-treated transgenic mice. When bone marrow from wild-type mice was transplanted into irradiated LysM DTR mice, the alveolar macrophages became resistant to DT but the mice still suffered from acute lung injury by DT administration. Compared with the mice in which both AE2 cells and macrophages were deleted by DT administration, the DT treated LysM-DTR mice with DT-resistant macrophages showed less severe lung injury with a reduced amount of hepatocyte growth factor in bronchoalveolar lavage fluid. These results indicate that macrophages play a protective role in noninflammatory lung injury caused by the selective ablation of AE2 cells. PMID- 17404283 TI - Germinal center helper T cells are dual functional regulatory cells with suppressive activity to conventional CD4+ T cells. AB - Germinal center (GC) reaction is a T cell-dependent process in which activated B cells mature to produce high-affinity Abs and differentiate into memory B cells. The GC microenvironment is almost exclusively reserved for the optimal Ag specific B cell clonal expansion, selection, and maturation, but lack significant conventional CD4(+) T cell responses. The mechanisms that ensure such a focused B cell response in the GC are not known. In this study, we report that human CD4(+)CD57(+) T cells, which are the major helper T cells in GCs, actively suppress the activation of conventional CD4(+) T cells, particularly Th1 cells, via a direct contact-dependent mechanism and soluble mediators. Our findings demonstrate that GC T cells are unique regulatory cells that provide critical help signals for B cell response but suppress conventional effector T cells in the same local environment. PMID- 17404284 TI - Severe defect in thymic development in an insertional mutant mouse model. AB - Transgenic mice were generated expressing NK1.1, an NK cell-associated receptor, under control of the human CD2 promoter. Unexpectedly, one of the founder lines, Tg66, showed a marked defect in thymic development characterized by disorganized architecture and small size. Mapping of the transgene insertion by fluorescence in situ hybridization revealed integration in chromosome 2, band G. Already from postnatal day 3, the thymic architecture was disturbed with a preferential loss of cortical thymic epithelial cells, a feature that became more pronounced over time. Compared with wild-type mice, total thymic cell numbers decreased dramatically between 10 and 20 days of age. Thymocytes isolated from adult Tg66 mice were predominantly immature double-negative cells, indicating a block in thymic development at an early stage of differentiation. Consequently, Tg66 mice had reduced numbers of peripheral CD4(+) and CD8(+) T cells. Bone marrow from Tg66 mice readily reconstituted thymi of irradiated wild-type as well as RAG deficient mice. This indicates that the primary defect in Tg66 mice resided in nonhemopoietic stromal cells of the thymus. The phenotype is observed in mice heterozygous for the insertion and does not resemble any known mutations affecting thymic development. Preliminary studies in mice homozygous for transgene insertion reveal a more accelerated and pronounced phenotype suggesting a semidominant effect. The Tg66 mice may serve as a useful model to identify genes regulating thymic epithelial cell differentiation, thymic development, and function. PMID- 17404285 TI - Beta-catenin regulates positive selection of thymocytes but not lineage commitment. AB - Positive selection and lineage commitment to the cytolytic or helper lineage of T cells result in coordinated expression of MHC class I-restricted TCR and CD8 coreceptor or MHC class II-restricted TCR and CD4 molecule. Positive selection signals also regulate the survival and generation of requisite numbers of cytolytic or Th cells. beta-Catenin is the major transcriptional cofactor of T cell factor and plays a role in thymocyte development. In this study, using mice expressing stabilized beta-catenin and mice with T cell-specific deletion of beta catenin, we show that beta-catenin regulates positive selection, but not lineage commitment of thymocytes. Furthermore, beta-catenin expression accelerates the timing of mature CD8 thymocyte generation such that CD4 and CD8 single-positive thymocytes mature with the same kinetics during development. PMID- 17404286 TI - Autoantigen-B cell antigen receptor interactions that regulate expression of B cell antigen receptor Loci. AB - Levels of AgR (BCR) expression are regulated during B cell development, activation, and induction of tolerance. The mechanisms responsible for and consequences of this regulation are poorly understood. We have described a class of DNA-based autoantigen-reactive B cell that down-regulates BCR expression during development to mature follicular phenotype. In this study, we show that at immature stages of primary differentiation, individual B cells of this type can dynamically modulate levels of expression of BCR in inverse proportion to degree of autoantigen engagement and induced BCR signaling. These adjustments in BCR expression are not associated with cell death, BCR revision, or altered development, and do not require TLR 9. Strikingly, modulation of BCR subunit gene RNA levels and transcription parallels these changes in BCR expression, indicating a direct link between autoantigen-BCR interactions of this type and regulation of transcription of BCR-encoding loci. We propose that this adaptive process allows this class of autoreactive B cell to avoid conventional tolerance pathways and promotes development to mature phenotype. PMID- 17404287 TI - A critical role for p53 in the control of NF-kappaB-dependent gene expression in TLR4-stimulated dendritic cells exposed to Genistein. AB - Considerable research has focused on the anti-inflammatory and antiproliferative activities exhibited by the soy isoflavone genistein. We previously demonstrated that genistein suppresses TNF-alpha-induced NF-kappaB-dependent IL-6 gene expression in cancer cells by interfering with the mitogen- and stress-activated protein kinase 1 activation pathway. However, effects of isoflavones on immune cells, such as dendritic cells, remain largely unknown. Here we show that genistein markedly reduces IL-6 cytokine production and transcription in LPS stimulated human monocyte-derived dendritic cells. More particularly, we observe that genistein inhibits IL-6 gene expression by modulating the transcription factor NF-kappaB. Examination of NF-kappaB-related events downstream of TLR4 demonstrates that genistein affects NF-kappaB subcellular localization and DNA binding, although we observe only a minor inhibitory impact of genistein on the classical LPS-induced signaling steps. Interestingly, we find that genistein significantly increases p53 protein levels. We also show that overexpression of p53 in TLR4/MD2 HEK293T cells blocks LPS-induced NF-kappaB-dependent gene transcription, indicating the occurrence of functional cross-talk between p53 and NF-kappaB. Moreover, analysis of IL-6 mRNA levels in bone marrow-derived p53 null vs wild-type dendritic cells confirms a role for p53 in the reduction of NF kappaB-dependent gene expression, mediated by genistein. PMID- 17404288 TI - Both the suppressor of cytokine signaling 1 (SOCS-1) kinase inhibitory region and SOCS-1 mimetic bind to JAK2 autophosphorylation site: implications for the development of a SOCS-1 antagonist. AB - Suppressor of cytokine signaling (SOCS)-1 protein modulates signaling by IFN gamma by binding to the autophosphorylation site of JAK2 and by targeting bound JAK2 to the proteosome for degradation. We have developed a small tyrosine kinase inhibitor peptide (Tkip) that is a SOCS-1 mimetic. Tkip is compared in this study with the kinase inhibitory region (KIR) of SOCS-1 for JAK2 recognition, inhibition of kinase activity, and regulation of IFN-gamma-induced biological activity. Tkip and a peptide corresponding to the KIR of SOCS-1, ((53))DTHFRTFRSHSDYRRI((68)) (SOCS1-KIR), both bound similarly to the autophosphorylation site of JAK2, JAK2(1001-1013). The peptides also bound to JAK2 peptide phosphorylated at Tyr(1007), pJAK2(1001-1013). Dose-response competitions suggest that Tkip and SOCS1-KIR similarly recognize the autophosphorylation site of JAK2, but probably not precisely the same way. Although Tkip inhibited JAK2 autophosphorylation as well as IFN-gamma-induced STAT1-alpha phosphorylation, SOCS1-KIR, like SOCS-1, did not inhibit JAK2 autophosphorylation but inhibited STAT1-alpha activation. Both Tkip and SOCS1-KIR inhibited IFN-gamma activation of Raw 264.7 murine macrophages and inhibited Ag specific splenocyte proliferation. The fact that SOCS1-KIR binds to pJAK2(1001 1013) suggests that the JAK2 peptide could function as an antagonist of SOCS-1. Thus, pJAK2(1001-1013) enhanced suboptimal IFN-gamma activity, blocked SOCS-1 induced inhibition of STAT3 phosphorylation in IL-6-treated cells, enhanced IFN gamma activation site promoter activity, and enhanced Ag-specific proliferation. Furthermore, SOCS-1 competed with SOCS1-KIR for pJAK2(1001-1013). Thus, the KIR region of SOCS-1 binds directly to the autophosphorylation site of JAK2 and a peptide corresponding to this site can function as an antagonist of SOCS-1. PMID- 17404289 TI - Description and mapping of the resistance of DBA/2 mice to TNF-induced lethal shock. AB - In our search for genes that inhibit the inflammatory effects of TNF without diminishing its antitumor capacities we found that, compared with C57BL/6 mice, DBA/2 mice exhibit a dominant resistance to TNF-induced lethality. Tumor-bearing (C57BL/6 x DBA/2)(BXD)F(1) mice completely survived an otherwise lethal TNF/IFN gamma-antitumor therapy with complete regression of the tumor. This was not the case for C57BL/6 mice. Genetic linkage analysis revealed that TNF resistance is linked to a major locus on distal chromosome 6 and a minor locus on chromosome 17. Compared with littermate controls, chromosome substitution mice carrying a DBA/2 chromosome 6 in a C57BL/6 background were significantly protected against TNF and TNF/IFN-gamma, albeit less so than DBA/2 mice. Definition of a critical region of 13 Mb on chromosome 6 was the highest mapping resolution obtained. Further analysis of candidate genes may provide a powerful tool to control TNF induced pathologies in humans. PMID- 17404290 TI - Genome-wide network analysis reveals the global properties of IFN-beta immediate transcriptional effects in humans. AB - IFN-beta effectively controls clinical exacerbations and magnetic resonance imaging activity in most multiple sclerosis patients. However, its mechanism of action has not been yet fully elucidated. In this study we used DNA microarrays to analyze the longitudinal transcriptional profile of blood cells within a week of IFN-beta administration. Using differential expression and gene ontology analyses we found evidence of a general decrease in the cellular activity of T lymphocytes resembling the endogenous antiviral response of IFNs. In contrast, most of the differentially expressed genes (DEGs) from untreated individuals were involved in cellular physiological processes. We then used mutual information (MI) to build networks of coregulated genes in both treated and untreated individuals. Interestingly, the connectivity distribution (k) of networks generated with high MI values displayed scale-free properties. Conversely, the observed k for networks generated with suboptimal MI values approximated a Poisson distribution, suggesting that MI captures biologically relevant interactions. Gene networks from individuals treated with IFN-beta revealed a tight core of immune- and apoptosis-related genes associated with higher values of MI. In contrast, networks obtained from untreated individuals primarily reflected cellular housekeeping functions. Finally, we trained a neural network to reverse engineer the directionality of the main interactions observed at the biological process level. This is the first study that incorporates network analysis to investigate gene regulation in response to a therapeutic drug in humans. Implications of this method in the creation of personalized models of response to therapy are discussed. PMID- 17404291 TI - IL-28 elicits antitumor responses against murine fibrosarcoma. AB - IL-28 is a recently described antiviral cytokine. In this study, we investigated the biological effects of IL-28 on tumor growth to evaluate its antitumor activity. IL-28 or retroviral transduction of the IL-28 gene into MCA205 cells did not affect in vitro growth, whereas in vivo growth of MCA205IL-28 was markedly suppressed along with survival advantages when compared with that of controls. When the metastatic ability of IL-28-secreting MCA205 cells was compared with that of controls, the expression of IL-28 resulted in a potent inhibition of metastases formation in the lungs. IL-28-mediated suppression of tumor growth was mostly abolished in irradiated mice, indicating that irradiation sensitive cells, presumably immune cells, are primarily involved in the IL-28 induced suppression of tumor growth. In vivo cell depletion experiments displayed that polymorphonuclear neutrophils, NK cells, and CD8 T cells, but not CD4 T cells, play an equal role in the IL-28-mediated inhibition of in vivo tumor growth. Consistent with these findings, inoculation of MCA205IL-28 into mice evoked enhanced IFN-gamma production and cytotoxic T cell activity in spleen cells. Antitumor action of IL-28 is partially dependent on IFN-gamma and is independent of IL-12, IL-17, and IL-23. IL-28 increased the total number of splenic NK cells in SCID mice and enhanced IL-12-induced IFN-gamma production in vivo and expanded spleen cells in C57BL/6 mice. Moreover, IL-12 augmented IL-28 mediated antitumor activity in the presence or absence of IFN-gamma. These findings indicate that IL-28 has bioactivities that induce innate and adaptive immune responses against tumors. PMID- 17404292 TI - A novel mode of action for a microbial-derived immunotoxin: the cytolethal distending toxin subunit B exhibits phosphatidylinositol 3,4,5-triphosphate phosphatase activity. AB - The Actinobacillus actinomycetemcomitans cytolethal distending toxin (Cdt) is a potent immunotoxin that induces G(2) arrest in human lymphocytes. We now show that the CdtB subunit exhibits phosphatidylinositol (PI)-3,4,5-triphosphate phosphatase activity. Breakdown product analysis indicates that CdtB hydrolyzes PI-3,4,5-P(3) to PI-3,4-P(2) and therefore functions in a manner similar to phosphatidylinositol 5-phosphatases. Conserved amino acids critical to catalysis in this family of enzymes were mutated in the cdtB gene. The mutant proteins exhibit reduced phosphatase activity along with decreased ability to induce G(2) arrest. Consistent with this activity, Cdt induces time-dependent reduction of PI 3,4,5-P(3) in Jurkat cells. Lymphoid cells with defects in SHIP1 and/or ptase and tensin homolog deleted on chromosome 10 (PTEN) (such as Jurkat, CEM, Molt) and, concomitantly, elevated PI-3,4,5-P(3) levels were more sensitive to the toxin than HUT78 cells which contain functional levels of both enzymes and low levels of PI-3,4,5-P(3). Finally, reduction of Jurkat cell PI-3,4,5-P(3) synthesis using the PI3K inhibitors, wortmannin and LY290004, protects cells from toxin-induced cell cycle arrest. Collectively, these studies show that the CdtB not only exhibits PI-3,4,5-P(3) phosphatase activity, but also that toxicity in lymphocytes is related to this activity. PMID- 17404293 TI - Increasing the recruitment of neutrophils to the site of infection dramatically attenuates Borrelia burgdorferi infectivity. AB - Borrelia burgdorferi infection causes an initial skin lesion called erythema migrans (EM) in human Lyme disease and in models of monkey and rabbit borreliosis. EM results from the inflammatory response triggered by spirochete replication and likely develops to contain the initial infection but allows bacterial dissemination to occur. The essential lack of neutrophil involvement in EM histopathology prompted us to examine the consequence of increasing their recruitment in the inflammatory response to the Lyme disease agent. B. burgdorferi was modified genetically to constitutively express and secrete the chemokine KC, a neutrophil chemoattractant. After inoculation into the dermis of the murine host, control spirochetes induced an infiltration of macrophages, neutrophils, and basophils within 6 h; however, the recruited neutrophils and basophils were quickly substituted by eosinophils, and the inflammatory response became macrophage dominant by 16 h. Such a response failed to contain the initial infection and allowed the spirochetes to disseminate. In contrast, B. burgdorferi with KC secretion induced an intensive neutrophil infiltration at the inoculation site, and as a result, the host's ability to control the initial infection was greatly enhanced. Taken together, this study suggests that the failure of sufficient neutrophil recruitment and activation during the initial inflammatory response may allow B. burgdorferi to effectively colonize the mammalian host. PMID- 17404294 TI - Mannose-binding lectin A-deficient mice have abrogated antigen-specific IgM responses and increased susceptibility to a nematode infection. AB - To investigate the role of mannose-binding lectin-A (MBL-A) in protection against infectious disease, MBL-A(-/-)-deficient mice were generated. Using a well characterized mouse model of human filarial nematode infection, nematode survival and protective immune responses were tested in vivo. Blood-borne Brugia malayi microfilariae survived for significantly longer time periods in MBL-A(-/-) than in wild-type (WT) mice. However, no differences in either splenic cytokine responses or induction of leukocytes in the blood were observed. A profound abrogation of Ag-specific IgM levels was measured in B. malayi-infected MBL-A(-/ ) mice, and some IgG isotypes were higher than those observed in WT animals. To establish whether there was a defect in Ab responses per se in MBL-A(-/-) mice or the effect was specific to filarial infection, we immunized these mice with OVA or a carbohydrate-free protein. Significantly, Ag-specific IgM responses were defective to both of these Ags, and Ag-specific IgG responses were largely unaffected. Furthermore, in naive mice, total IgM levels did not differ between MBL-A(-/-) and WT mice. This article describes the first demonstration that MBL-A may function independently of MBL-C and suggests that MBL-A, like other C-type lectins and members of the complement cascade, is intimately involved in the priming of the humoral Ab response. PMID- 17404295 TI - MyD88 is required for the formation of long-term humoral immunity to virus infection. AB - Development of long-term humoral immunity is a major goal of vaccination, but the mechanisms involved in the formation of long-term Ab responses are still being determined. In this study, we identify a previously unknown requirement for MyD88, an adaptor molecule that mediates signals at most TLRs, for the generation of long-term humoral immunity during live virus infection. Polyoma virus-infected MyD88 knockout mice generated strong acute T cell-dependent antiviral IgM and IgG responses and developed germinal centers. Activation-induced cytidine deaminase, an enzyme required for isotype switching and somatic hypermutation, was also induced in germinal center B cells, similar to wild-type mice. However, MyD88 knockout mice failed to develop bone marrow plasma cells and did not maintain long-term serum antiviral Ab responses. The isotype distribution of antiviral IgG responses was also altered; serum IgG2a and IgG2b levels were diminished, whereas IgG1 responses were not affected. The requirement for MyD88 for the formation of long-term humoral immunity to polyoma virus was intrinsic to B cells and was independent of IL-1R and IL-18R, cytokine receptors that also signal through MyD88. Our findings show that MyD88-dependent signaling pathways in B cells are essential for effectively generating long-term Ab responses and implicate a role for TLR in the formation of long-term humoral immunity. PMID- 17404296 TI - Impaired host defense in mice lacking ONZIN. AB - ONZIN is a small, cysteine-rich peptide of unique structure that is conserved in all vertebrates examined to date. We show that ONZIN is expressed at high levels in epithelial cells of the intestinal tract, the lung, and in cells of the immune system including macrophages and granulocytes. Because this pattern of expression is suggestive of a role in innate immune function, we have generated mice lacking this protein and examined their ability to respond to challenge with infectious agents. Onzin(-/-) mice show a heightened innate immune response after induction of acute peritonitis with Klebsiella pneumoniae. This increased response is consistent with an increased bacterial burden in the Onzin(-/-) mice. Ex vivo studies show that, whereas phagocytosis is not altered in Onzin(-/-) neutrophils, phagocytes lacking this protein kill bacteria less effectively. This result identifies ONZIN as a novel class of intracellular protein required for optimal function of the neutrophils after uptake of bacteria. PMID- 17404297 TI - Thiocyanate transport in resting and IL-4-stimulated human bronchial epithelial cells: role of pendrin and anion channels. AB - SCN(-) (thiocyanate) is an important physiological anion involved in innate defense of mucosal surfaces. SCN(-) is oxidized by H(2)O(2), a reaction catalyzed by lactoperoxidase, to produce OSCN(-) (hypothiocyanite), a molecule with antimicrobial activity. Given the importance of the availability of SCN(-) in the airway surface fluid, we studied transepithelial SCN(-) transport in the human bronchial epithelium. We found evidence for at least three mechanisms for basolateral to apical SCN(-) flux. cAMP and Ca(2+) regulatory pathways controlled SCN(-) transport through cystic fibrosis transmembrane conductance regulator and Ca(2+)-activated Cl(-) channels, respectively, the latter mechanism being significantly increased by treatment with IL-4. Stimulation with IL-4 also induced the strong up-regulation of an electroneutral SCN(-)/Cl(-) exchange. Global gene expression analysis with microarrays and functional studies indicated pendrin (SLC26A4) as the protein responsible for this SCN(-) transport. Measurements of H(2)O(2) production at the apical surface of bronchial cells indicated that the extent of SCN(-) transport is important to modulate the conversion of this oxidant molecule by the lactoperoxidase system. Our studies indicate that the human bronchial epithelium expresses various SCN(-) transport mechanisms under resting and stimulated conditions. Defects in SCN(-) transport in the airways may be responsible for susceptibility to infections and/or decreased ability to scavenge oxidants. PMID- 17404298 TI - Toxoplasma gondii dysregulates IFN-gamma-inducible gene expression in human fibroblasts: insights from a genome-wide transcriptional profiling. AB - Toxoplasma gondii is an obligate intracellular parasite that persists for the life of a mammalian host. The parasite's ability to block the potent IFN-gamma response may be one of the key mechanisms that allow Toxoplasma to persist. Using a genome-wide microarray analysis, we show here a complete dysregulation of IFN gamma-inducible gene expression in human fibroblasts infected with Toxoplasma. Notably, 46 of the 127 IFN-gamma-responsive genes were induced and 19 were suppressed in infected cells before they were exposed to IFN-gamma, indicating that other stimuli produced during infection may also regulate these genes. Following IFN-gamma treatment, none of the 127 IFN-gamma-responsive genes could be significantly induced in infected cells. Immunofluorescence assays showed at single-cell levels that infected cells, regardless of which Toxoplasma strain was used, could not be activated by IFN-gamma to up-regulate the expression of IFN regulatory factor 1, a transcription factor that is under the direct control of STAT1, whereas uninfected cells in the same culture expressed IFN regulatory factor 1 normally in response to IFN-gamma. STAT1 trafficked to the nucleus normally and indistinguishably in all uninfected and infected cells treated with IFN-gamma, indicating that the inhibitory effects of Toxoplasma infection likely occur via blocking STAT1 transcriptional activity in the nucleus. In contrast, a closely related apicomplexan, Neospora caninum, was unable to inhibit IFN-gamma induced gene expression. A differential ability to interfere with the IFN-gamma response may, in part, account for the differences in the pathogenesis seen among Toxoplasma and Neospora parasite strains. PMID- 17404300 TI - Role of MyD88 in route-dependent susceptibility to vesicular stomatitis virus infection. AB - TLRs are important components of the innate immune response. The role of the TLR signaling pathway in host defense against a natural viral infection has been largely unexplored. We found that mice lacking MyD88, an essential adaptor protein in TLR signaling pathway, were extremely sensitive to intranasal infection with vesicular stomatitis virus, and this susceptibility was dose dependent. We demonstrated that this increased susceptibility correlates with the impaired production of IFN-alpha and defective induction and maintenance of neutralizing Ab. These studies outline the important role of the TLR signaling pathway in nasal mucosae-respiratory tracts-neuroepithelium environment in the protection against microbial pathogen infections. We believe that these results explain how the route of infection, probably by virtue of activating different cell populations, can lead to entirely different outcomes of infection based on the underlying genetics of the host. PMID- 17404299 TI - Oligoadenylate synthetase/protein kinase R pathways and alphabeta TCR+ T cells are required for adenovirus vector: IFN-gamma inhibition of herpes simplex virus 1 in cornea. AB - An adenoviral (Ad) vector containing the murine IFN-gamma transgene (Ad:IFN gamma) was evaluated for its capacity to inhibit HSV-1. To measure effectiveness, viral titers were analyzed in cornea and trigeminal ganglia (TG) during acute ocular HSV-1 infection. Ad:IFN-gamma potently suppressed HSV-1 replication in a dose-dependent fashion, requiring IFN-gamma receptor. Moreover, Ad:IFN-gamma was effective when delivered -72 and -24 h before infection as well as 24 h postinfection. Associated with antiviral opposition, TG from Ad:IFN-gamma transduced mice harbored fewer T cells. Also related to T cell involvement, Ad:IFN-gamma was effective but attenuated in TG from alphabeta TCR-deficient mice. In corneas, alphabeta TCR(+) T cells were obligatory for protection against viral multiplication. Type I IFN involvement amid antiviral efficacy of Ad:IFN gamma was further investigated because types I and II IFN pathways have synergistic anti-HSV-1 activity. Ad:IFN-gamma inhibited viral reproduction in corneas and TG from alphabeta IFNR-deficient (CD118(-/-)) mice, although viral titers were 2- to 3-fold higher in cornea and TG compared with wild-type mice. The absence of IFN-stimulated antiviral proteins, 2'-5' oligoadenylate synthetase/RNase L, and dsRNA-dependent protein kinase R completely eliminated the antiviral effectiveness of Ad:IFN-gamma. Collectively, the results demonstrate the following: 1) nonexistence of type I IFN receptor does not abolish defense of Ad:IFN-gamma against HSV-1; 2) antiviral pathways oligoadenylate synthetase-RNase L and protein kinase R are mandatory; and 3) alphabeta TCR(+) T cells are compulsory for Ad:IFN-gamma effectiveness against HSV-1 in cornea but not in TG. PMID- 17404301 TI - MyD88-dependent activation of B220-CD11b+LY-6C+ dendritic cells during Brucella melitensis infection. AB - IFN-gamma is a key cytokine controlling Brucella infection. One of its major function is the stimulation of Brucella-killing effector mechanisms, such as inducible NO synthase (iNOS)/NOS2 activity, in phagocytic cells. In this study, an attempt to identify the main cellular components of the immune response induced by Brucella melitensis in vivo is made. IFN-gamma and iNOS protein were analyzed intracellularly using flow cytometry in chronically infected mice. Although TCRbeta(+)CD4(+) cells were the predominant source of IFN-gamma in the spleen, we also identified CD11b(+)LY-6C(+)LY-6G(-)MHC-II(+) cells as the main iNOS-producing cells in the spleen and the peritoneal cavity. These cells appear similar to inflammatory dendritic cells recently described in the mouse model of Listeria monocytogenes infection and human psoriasis: the TNF/iNOS-producing dendritic cells. Using genetically deficient mice, we demonstrated that the induction of iNOS and IFN-gamma-producing cells due to Brucella infection required TLR4 and TLR9 stimulation coupled to Myd88-dependent signaling pathways. The unique role of MyD88 was confirmed by the lack of impact of Toll-IL-1R domain containing adaptor inducing IFN-beta deficiency. The reduction of IFN-gamma(+) and iNOS(+) cell frequency observed in MyD88-, TLR4-, and TLR9-deficient mice correlated with a proportional lack of Brucella growth control. Taken together, our results provide new insight into how immune responses fight Brucella infection. PMID- 17404302 TI - Disparity in IL-12 release in dendritic cells and macrophages in response to Mycobacterium tuberculosis is due to use of distinct TLRs. AB - The control of IL-12 production from dendritic cells (DCs) and macrophages in response to Mycobacterium tuberculosis (Mtb) is not well understood. The objective of this study was to pursue the mechanism underlying our previous report that in response to Mtb infection, DCs release abundant IL-12, whereas secretion is limited in macrophages. An initial comparison of IL-12p35 and IL 12p40 gene induction showed that p35 transcription is similar in murine bone marrow-derived DCs and macrophages, but a rapid and enhanced IL-12p40 transcription occurs only in DCs. Consistent with the p40 gene transcription profile, Mtb-induced remodeling at nucleosome 1 of the p40 promoter also occurs rapidly and extensively in DCs in comparison to macrophages. Removal of IL-10 or addition of IFNgamma enhances macrophage IL-12 release to Mtb, but without affecting the kinetics of remodeling at the macrophage p40 promoter. Furthermore, we show that Mtb-induced remodeling at the p40 promoter and IL-12 release in DCs is TLR9 dependent, and in contrast, TLR2 dependent, in macrophages. Data are also presented to demonstrate that a TLR9 agonist induces quantitatively more extensive remodeling at the IL-12p40 promoter and larger IL-12 release in comparison to a TLR2 agonist. Collectively, these findings suggest that DCs and macrophages handle Mtb differently resulting in only DCs being able to engage the more efficient TLR9 pathway for IL-12 gene induction. Our results also imply that TLR2 signaling is not a good inducer of IL-12, supporting the increasingly strong paradigm that TLR2 favors Th2 responses. PMID- 17404303 TI - TLR3 is essential for the induction of protective immunity against Punta Toro Virus infection by the double-stranded RNA (dsRNA), poly(I:C12U), but not Poly(I:C): differential recognition of synthetic dsRNA molecules. AB - In the wake of RNA virus infections, dsRNA intermediates are often generated. These viral pathogen-associated molecular patterns can be sensed by a growing number of host cell cytosolic proteins and TLR3, which contribute to the induction of antiviral defenses. Recent evidence indicates that melanoma differentiation-associated gene-5 is the prominent host component mediating IFN production after exposure to the dsRNA analog, poly(I:C). We have previously reported that Punta Toro virus (PTV) infection in mice is exquisitely sensitive to treatment with poly(I:C(12)U), a dsRNA analog that has a superior safety profile while maintaining the beneficial activity of the parental poly(I:C) in the induction of innate immune responses. The precise host factor(s) mediating protective immunity following its administration remain to be elucidated. To assess the role of TLR3 in this process, mice lacking the receptor were used to investigate the induction of protective immunity, type I IFNs, and IL-6 following treatment. Unlike wild-type mice, those lacking TLR3 were not protected against PTV infection following poly(I:C(12)U) therapy and failed to produce IFN-alpha, IFN-beta, and IL-6. In contrast, poly(I:C) treatment significantly protected TLR3(-/-) mice from lethal challenge despite some deficiencies in cytokine induction. There was no indication that the lack of protection was due to the fact that TLR3-deficient mice had a reduced capacity to fight infection because they were not found to be more susceptible to PTV. We conclude that TLR3 is essential to the induction of antiviral activity elicited by poly(I:C(12)U), which does not appear to be recognized by the cytosolic sensor of poly(I:C), melanoma differentiation-associated gene-5. PMID- 17404304 TI - Role of the indigenous microbiota in maintaining the virus-specific CD8 memory T cells in the lung of mice infected with murine cytomegalovirus. AB - The potent role of indigenous microbiota in maintaining murine CMV (MCMV) specific memory T cells, which were measured by multimer staining, was investigated using germfree (GF) mice. When the BALB/c mice bred under specific pathogen-free (SPF) conditions were i.p. infected with 0.2 LD(50) of MCMV, high frequencies of CD69(+)/CD44(+) MCMV-specific CD8 T cells were noted in the lungs even at 6-12 mo after infection (11.1 +/- 3.2 and 9.8 +/- 0.9%, respectively). In contrast, even though the viral load and expression levels of mRNA of such cytokines as IL-2, IL-7, IL-15, and IFN-gamma in the lungs of MCMV-infected GF mice were comparable to those of infected SPF mice, the frequencies of MCMV specific CD8 T cells in the lungs of infected GF mice were kept lower than 1% at 6-12 mo after infection. In addition, the reconstitution of microbiota of MCMV infected GF mice by orally administering a fecal suspension prepared from SPF mice restored the frequencies of both CD8(+)/multimer(+) and CD8(+)/multimer(-) T cells to levels similar to those found in SPF mice. These results suggested the indigenous microbiota to play a crucial role in the expansion and maintenance of viral-specific CD8 memory T cells, probably by cross-reactivity between the antigenic epitope of the MCMV-specific memory T cells and the variety of peptides derived from the members of the microbiota. Such cross-reactivity may thus be a major feature of those cells. PMID- 17404305 TI - Dynamic relationship between IFN-gamma and IL-2 profile of Mycobacterium tuberculosis-specific T cells and antigen load. AB - Distinct IFN-gamma and IL-2 profiles of Ag-specific CD4(+) T cells have recently been associated with different clinical disease states and Ag loads in viral infections. We assessed the kinetics and functional profile of Mycobacterium tuberculosis Ag-specific T cells secreting IFN-gamma and IL-2 in 23 patients with untreated active tuberculosis when bacterial and Ag loads are high and after curative treatment, when Ag load is reduced. The frequencies of M. tuberculosis Ag-specific IFN-gamma-secreting T cells declined during 28 mo of follow-up with an average percentage decline of 5.8% per year (p = 0.005), while the frequencies of Ag-specific IL-2-secreting T cells increased during treatment (p = 0.02). These contrasting dynamics for the two cytokines led to a progressive convergence of the frequencies of IFN-gamma- and IL-2-secreting cells over 28 mo. Simultaneous measurement of IFN-gamma and IL-2 secretion at the single-cell level revealed a codominance of IFN-gamma-only secreting and IFN-gamma/IL-2 dual secreting CD4(+) T cells in active disease that shifted to dominance of IFN gamma/IL-2-secreting CD4(+) T cells and newly detectable IL-2-only secreting CD4(+) T cells during and after treatment. These distinct T cell functional signatures before and after treatment suggest a novel immunological marker of mycobacterial load and clinical status in tuberculosis that now requires validation in larger prospective studies. PMID- 17404306 TI - CD8+ T cell dysfunction and increase in murine gammaherpesvirus latent viral burden in the absence of 4-1BB ligand. AB - Studies of costimulatory receptors belonging to the TNFR family have revealed their diverse roles in affecting different stages of the T cell response. The 4 1BB ligand (4-1BBL)/4-1BB pathway has emerged as a receptor-ligand pair that impacts not the initial priming, but later phases of the T cell response, such as sustaining clonal expansion and survival, maintaining memory CD8(+) T cells, and supporting secondary expansion upon Ag challenge. Although the role of this costimulatory pathway in CD8(+) T cell responses to acute viral infections has been well-studied, its role in controlling chronic viral infections in vivo is not known to date. Using the murine gammaherpesvirus-68 (MHV-68) model, we show that 4-1BBL-deficient mice lack control of MHV-68 during latency and show significantly increased latent viral loads. In contrast to acute influenza infection, the numbers of MHV-68-specific memory CD8(+) T cells were maintained during latency. However, the virus-specific CD8(+) T cells showed defects in function, including decreased cytolytic function and impaired secondary expansion. Thus, 4-1BBL deficiency significantly affects the function, but not the number, of virus-specific CD8(+) T cells during gammaherpesvirus latency, and its absence results in an increased viral burden. Our study suggests that the 4 1BB costimulatory pathway plays an important role in controlling chronic viral infections. PMID- 17404307 TI - Mold allergen, pen C 13, induces IL-8 expression in human airway epithelial cells by activating protease-activated receptor 1 and 2. AB - Allergenic serine proteases are important in the pathogenesis of asthma. One of these, Pen c 13, is the immunodominant allergen produced by Penicillium citrinum. Many serine proteases induce cytokine expression, but whether Pen c 13 does so in human respiratory epithelial cells is not known. In this study, we investigated whether Pen c 13 caused IL-8 release and activated protease-activated receptors (PARs) in airway epithelial cells. In airway-derived A549 cells and normal human airway epithelial cells, Pen c 13 induced IL-8 release in a dose-dependent manner. Pen c 13 also increased IL-8 release in a time-dependent manner in A549 cells. Pen c 13 cleaved PAR-1 and PAR-2 at their activation sites. Treatment with Pen c 13 induced intracellular Ca(2+) mobilization and desensitized the cells to the action of other proteases and PAR-1 and PAR-2 agonists. Moreover, Pen c 13 mediated IL-8 release was significantly decreased in Ca(2+)-free medium and was abolished by the protease inhibitors, PMSF and 4-(2-aminoethyl) benzenesulfonyl fluoride. Blocking Abs against the cleavage sites of PAR-1 and PAR-2, but not of PAR-4, inhibited Pen c 13-induced IL-8 production, as did inhibition of phospholipase C. Pen c 13 induced IL-8 expression via activation of ERK 1/2, and not of p38 and JNK. In addition, treatment of A549 cells or normal human airway epithelial cells with Pen c 13 increased phosphorylation of ERK 1/2 by a Ca(2+) dependent pathway. These finding show that Pen c 13 induces IL-8 release in airway epithelial cells and that this is dependent on PAR-1 and PAR-2 activation and intracellular calcium. PMID- 17404308 TI - Granulocyte-macrophage colony-stimulating factor (CSF) and macrophage CSF dependent macrophage phenotypes display differences in cytokine profiles and transcription factor activities: implications for CSF blockade in inflammation. AB - GM-CSF and M-CSF (CSF-1) can enhance macrophage lineage numbers as well as modulate their differentiation and function. Of recent potential significance for the therapy of inflammatory/autoimmune diseases, their blockade in relevant animal models leads to a reduction in disease activity. What the critical actions are of these CSFs on macrophages during inflammatory reactions are unknown. To address this issue, adherent macrophages (GM-BMM and BMM) were first derived from murine bone marrow precursors by GM-CSF and M-CSF, respectively, and stimulated in vitro with LPS to measure secreted cytokine production, as well as NF-kappaB and AP-1 activities. GM-BMM preferentially produced TNF-alpha, IL-6, IL-12p70, and IL-23 whereas, conversely, BMM generated more IL-10 and CCL2; strikingly the latter population could not produce detectable IL-12p70 and IL-23. Following LPS stimulation, GM-BMM displayed rapid IkappaBalpha degradation, RelA nuclear translocation, and NF-kappaB DNA binding relative to BMM, as well as a faster and enhanced AP-1 activation. Each macrophage population was also pretreated with the other CSF before LPS stimulation and found to adopt the phenotype of the other population to some extent as judged by cytokine production and NF-kappaB activity. Thus, GM-CSF and M-CSF demonstrate, at the level of macrophage cytokine production, different and even competing responses with implications for their respective roles in inflammation, including a possible dampening or suppressive role for M-CSF in certain circumstances. PMID- 17404309 TI - Free cholesterol alters lipid raft structure and function regulating neutrophil Ca2+ entry and respiratory burst: correlations with calcium channel raft trafficking. AB - Recent studies associate cholesterol excess and atherosclerosis with inflammation. The link between these processes is not understood, but cholesterol is an important component of lipid rafts. Rafts are thought to concentrate membrane signaling molecules and thus regulate cell signaling through G protein coupled pathways. We used methyl beta-cyclodextrin to deplete cholesterol from polymorphonuclear neutrophil (PMN) rafts and thus study the effects of raft disruption on G protein-coupled Ca(2+) mobilization. Methyl beta-cyclodextrin had no effect on Ca(2+) store depletion by the G protein-coupled agonists platelet activating factor or fMLP, but abolished agonist-stimulated Ca(2+) entry. Free cholesterol at very low concentrations regulated Ca(2+) entry into PMN via nonspecific Ca(2+) channels in a biphasic fashion. The specificity of cholesterol regulation for Ca(2+) entry was confirmed using thapsigargin studies. Responses to cholesterol appear physiologic because they regulate respiratory burst in a proportional biphasic fashion. Investigating further, we found that free cholesterol accumulated in PMN lipid raft fractions, promoting formation and polarization of membrane rafts. Finally, the transient receptor potential calcium channel protein TRPC1 redistributed to raft fractions in response to cholesterol. The uniformly biphasic relationships between cholesterol availability, Ca(2+) signaling and respiratory burst suggest that Ca(2+) influx and PMN activation are regulated by the quantitative relationships between cholesterol and other environmental lipid raft components. The association between symptomatic cholesterol excess and inflammation may therefore in part reflect free cholesterol- dependent changes in lipid raft structure that regulate immune cell Ca(2+) entry. Ca(2+) entry-dependent responses in other cell types may also reflect cholesterol bioavailability and lipid incorporation into rafts. PMID- 17404310 TI - IFN-gamma induces cysteinyl leukotriene receptor 2 expression and enhances the responsiveness of human endothelial cells to cysteinyl leukotrienes. AB - Cysteinyl leukotrienes (cysLTs) are important mediators of cell trafficking and innate immune responses, involved in the pathogenesis of inflammatory processes, i.e., atherosclerosis, pulmonary fibrosis, and bronchial asthma. The aim of this study was to examine the regulation of cysLT signaling by IFN-gamma in human primary endothelial cells. IFN-gamma increased cysLT receptor 2 (CysLTR2) mRNA expression and CysLTR2-specific calcium signaling in endothelial cells. IFN-gamma signaled through Jak/STAT1, as both AG490, a Jak2 inhibitor, and expression of a STAT1 dominant-negative construct, significantly inhibited CysLTR2 mRNA expression in response to IFN-gamma. To determine mechanisms of IFN-gamma-induced CysLTR2 expression, the human CysLTR2 gene structure was characterized. The CysLTR2 gene has a TATA-less promoter, with multiple transcription start sites. It consists of six variably spliced exons. Eight different CysLTR2 transcripts were identified in endothelial and monocytic cells. Gene reporter assay showed potent basal promoter activity of a putative CysLTR2 promoter region. However, there were no significant changes in gene reporter and mRNA t(1/2) assays in response to IFN-gamma, suggesting transcriptional control of CysLTR2 mRNA up regulation by IFN-gamma response motifs localized outside of the cloned CysLTR2 promoter region. Stimulation of endothelial cells by cysLTs induced mRNA and protein expression of early growth response genes 1, 2, and 3 and cycloxygenase 2. This response was mediated by CysLTR2 coupled to G(q/11), activation of phospholipase C, and inositol-1,4,5-triphosphate, and was enhanced further 2- to 5-fold by IFN-gamma stimulation. Thus, IFN-gamma induces CysLTR2 expression and enhances cysLT-induced inflammatory responses. PMID- 17404311 TI - Aluminum hydroxide adjuvants activate caspase-1 and induce IL-1beta and IL-18 release. AB - Aluminum hydroxide (Alum) is the only adjuvant approved for routine use in humans, although the basis for its adjuvanticity remains poorly understood. In this study, we show that Alum activates caspase-1 and induce secretion of mature IL-1beta and IL-18. Human PBMC or dendritic cells stimulated with pure TLR4 and TLR2 agonists released only traces of IL-1beta or IL-18, despite the fact that the IL-1beta mRNA was readily induced by both TLR agonists. In contrast, cells costimulated with TLR agonists plus Alum released large amount of IL-1beta and IL 18. Alum-induced IL-1beta and IL-18 production was not due to enhancement of TLR signaling but rather reflected caspase-1 activation and in mouse dendritic cells occurred in a MyD88-independent fashion. Secretion of other proinflammatory cytokines such as IL-8 was not affected by Alum treatments. However, TLR-induced production of IL-10 was increased and that of IFN-gamma-inducible protein decreased by Alum cotreatment. Considering the immunostimulatory activities of these cytokines and the ability of IL-1beta to act as adjuvant, our results suggest a mechanism for the adjuvanticity of Alum. PMID- 17404313 TI - Scar-associated macrophages are a major source of hepatic matrix metalloproteinase-13 and facilitate the resolution of murine hepatic fibrosis. AB - Both the identity and source of the rodent collagenase(s) that mediates matrix remodeling in liver fibrosis remain elusive. We have recently demonstrated an unequivocal role for scar-associated macrophages (SAMs) in the spontaneous resolution of liver fibrosis and sought to determine whether SAMs are the source of matrix metalloproteinase (MMP) 13 (collagenase 3), considered to be the primary interstitial collagenase in rodents. In this study, we demonstrate an association between MMP13 expression and the presence of SAMs in the regression of experimental liver fibrosis. mmp13 gene expression was restricted to regions of fibrosis that were rich in SAMs. Both MMP13 mRNA and protein colocalized to large phagocytes within and directly apposed to hepatic scars. Using the CD11b DTR-transgenic mouse to deplete SAMs in a model of chronic CCl(4) injury, we found that SAM depletion resulted in a 5-fold reduction in mmp13 message (p = 0.005). Furthermore, resolution of CCl(4)-induced fibrosis was retarded in MMP13 deficient mice. Thus, SAMs selectively, during resolution of fibrosis induce and use the major collagenase MMP13 to mediate the resorption of interstitial matrix and successfully remodel the fibrotic liver. PMID- 17404312 TI - Inflammatory reactive oxygen species-mediated hemopoietic suppression in Fancc deficient mice. AB - Patients with the genomic instability syndrome Fanconi anemia (FA) commonly develop progressive bone marrow (BM) failure and have a high risk of cancer. Certain manifestations of the disease suggest that the FA immune system is dysfunctional and may contribute to the pathogenesis of both BM failure and malignancies. In this study, we have investigated inflammation and innate immunity in FA hemopoietic cells using mice deficient in Fanconi complementation group C gene (Fancc). We demonstrate that Fancc-deficient mice exhibit enhanced inflammatory response and are hypersensitive to LPS-induced septic shock as a result of hemopoietic suppression. This exacerbated inflammatory phenotype is intrinsic to the hemopoietic system and can be corrected by the re-expression of a wild-type FANCC gene, suggesting a potential role of the FANCC protein in innate immunity. LPS-mediated hemopoietic suppression requires two major inflammatory agents, TNF-alpha and reactive oxygen species. In addition, LPS induced excessive accumulation of reactive oxygen species in Fancc(-/-) BM cells overactivates the stress kinase p38 and requires prolonged activation of the JNK. Our data implicate a role of inflammation in pathogenesis of FA and BM failure diseases in general. PMID- 17404314 TI - Inhibition of CCL1-CCR8 interaction prevents aggregation of macrophages and development of peritoneal adhesions. AB - Peritoneal adhesions are a significant complication of surgery and visceral inflammation; however, the mechanism has not been fully elucidated. The aim of this study was to clarify the mechanism of peritoneal adhesions by focusing on the cell trafficking and immune system in the peritoneal cavity. We investigated the specific recruitment of peritoneal macrophages (PMphi) and their expression of chemokine receptors in murine models of postoperative and postinflammatory peritoneal adhesions. PMphi aggregated at the site of injured peritoneum in these murine models of peritoneal adhesions. The chemokine receptor CCR8 was up regulated in the aggregating PMphi when compared with naive PMphi. The up regulation of CCR8 was also observed in PMphi, but not in bone marrow-derived Mphi, treated with inflammatory stimulants including bacterial components and cytokines. Importantly, CCL1, the ligand for CCR8, a product of both PMphi and peritoneal mesothelial cells (PMCs) following inflammatory stimulation, was a potent enhancer of CCR8 expression. Cell aggregation involving PMphi and PMCs was induced in vitro in the presence of CCL1. CCL1 also up-regulated mRNA levels of plasminogen activator inhibitor-1 in both PMphi and PMCs. CCR8 gene-deficient mice or mice treated with anti-CCL1-neutralizing Ab exhibited significantly reduced postoperational peritoneal adhesion. Our study now establishes a unique autocrine activation system in PMphi and the mechanism for recruitment of PMphi together with PMCs via CCL1/CCR8, as immune responses of peritoneal cavity, which triggers peritoneal adhesions. PMID- 17404315 TI - Chemokine receptor CCR2 but not CCR5 or CCR6 mediates the increase in pulmonary dendritic cells during allergic airway inflammation. AB - Increased numbers of pulmonary dendritic cells (DCs) are recruited to the lungs during allergic airway inflammation and contribute to the maintenance of the inflammatory immune response. The chemokine receptors that directly control DC accumulation into the lungs are largely unknown. To explore this issue, we generated mixed bone marrow chimeric mice containing both wild-type and knockout cells for a given chemokine receptor. After induction of allergic airway inflammation, we specifically tracked and compared chemokine receptor knockout vs wild-type DC populations through various lung compartments. Using this approach, we show that CCR2, but not CCR5 or CCR6, directly controls the accumulation of DCs into allergic lungs. Furthermore, the size of inflammatory monocyte populations in peripheral blood was strikingly CCR2 dependent, suggesting that CCR2 primarily mediates the release of monocytic DC precursors into the bloodstream. PMID- 17404316 TI - Knockout of Mkp-1 enhances the host inflammatory responses to gram-positive bacteria. AB - MAPK phosphatase (MKP)-1 is an archetypal member of the dual specificity protein phosphatase family that dephosphorylates MAPK. We have previously demonstrated that MKP-1 acts as a negative regulator of p38 and JNK in immortalized macrophages after stimulation with peptidoglycan isolated from Gram-positive bacteria. To define the physiological function of MKP-1 during Gram-positive bacterial infection, we studied the innate immune responses to Gram-positive bacteria using Mkp-1 knockout (KO) mice. We found that Mkp-1(-/-) macrophages exhibited prolonged activation of p38 and JNK, but not of ERK, following exposure to either peptidoglycan or lipoteichoic acid. Compared with wild-type (WT) macrophages, Mkp-1(-/-) macrophages produced more proinflammatory cytokines such as TNF-alpha and IL-6. Moreover, after challenge with peptidoglycan, lipoteichoic acid, live or heat-killed Staphylococcus aureus bacteria, Mkp-1 KO mice also mounted a more robust production of cytokines and chemokines, including TNF alpha, IL-6, IL-10, and MIP-1alpha, than did WT mice. Accordingly, Mkp-1 KO mice also exhibited greater NO production, more robust neutrophil infiltration, and more severe organ damage than did WT mice. Surprisingly, WT and Mkp-1 KO mice exhibited no significant difference in either bacterial load or survival rates when infected with live S. aureus. However, in response to challenge with heat killed S. aureus, Mkp-1 KO mice exhibited a substantially higher mortality rate compared with WT mice. Our studies indicate that MKP-1 plays a critical role in the inflammatory response to Gram-positive bacterial infection. MKP-1 serves to limit the inflammatory reaction by inactivating JNK and p38, thus preventing multiorgan failure caused by exaggerated inflammatory responses. PMID- 17404317 TI - Coexposure to environmental tobacco smoke increases levels of allergen-induced airway remodeling in mice. AB - Environmental tobacco smoke (ETS) can increase asthma symptoms and the frequency of asthma attacks. However, the contribution of ETS to airway remodeling in asthma is at present unknown. In this study, we have used a mouse model of allergen-induced airway remodeling to determine whether the combination of chronic exposure to ETS and chronic exposure to OVA allergen induces greater levels of airway remodeling than exposure to either chronic ETS or chronic OVA allergen alone. Mice exposed to chronic ETS alone did not develop significant eosinophilic airway inflammation, airway remodeling, or increased airway hyperreactivity to methacholine. In contrast, mice exposed to chronic OVA allergen had significantly increased levels of peribronchial fibrosis, increased thickening of the smooth muscle layer, increased mucus, and increased airway hyperreactivity which was significantly enhanced by coexposure to the combination of chronic ETS and chronic OVA allergen. Mice coexposed to chronic ETS and chronic OVA allergen had significantly increased levels of eotaxin-1 expression in airway epithelium which was associated with increased numbers of peribronchial eosinophils, as well as increased numbers of peribronchial cells expressing TGF beta1. These studies suggest that chronic coexposure to ETS significantly increases levels of allergen-induced airway remodeling (in particular smooth muscle thickness) and airway responsiveness by up-regulating expression of chemokines such as eotaxin-1 in airway epithelium with resultant recruitment of cells expressing TGF-beta1 to the airway and enhanced airway remodeling. PMID- 17404318 TI - The lymphocytic infiltration in calcific aortic stenosis predominantly consists of clonally expanded T cells. AB - Valve lesions in degenerative calcific aortic stenosis (CAS), a disorder affecting 3% of those older than 75 years, are infiltrated by T lymphocytes. We sought to determine whether the alphabeta TCR repertoire of these valve infiltrating lymphocytes exhibited features either of a polyclonal nonselective response to inflammation or contained expanded clones suggesting a more specific immune process. TCR beta-chain CDR3-length distribution analysis using PCR primers specific for 23 Vbeta families performed in eight individuals with CAS affecting tri- or bileaflet aortic valves revealed considerable oligoclonal T cell expansion. In five cases, beta-chain nucleotide sequencing in five selected Vbeta families showed that an average of 92% of the valve-infiltrating T cell repertoire consisted of expanded T cell clones, differing markedly in composition from the relatively more polyclonal peripheral CD8 or CD4 T cell subsets found even in this elderly population. Twenty-four of the valve-infiltrating T cell clones also had the same clone identified in blood, some of which were highly expanded. Interestingly, 22 of these 24 shared clones were CD8 in lineage (p = 1.5 x 10(-12)), suggesting a possible relationship to the expanded CD8(+)CD28(-) T cell clones frequently present in the elderly. Additionally, the sequences of several TCR beta-chain CDR3 regions were homologous to TCR beta-chains identified previously in allograft arteriosclerosis. We infer that these findings are inconsistent with a nonselective secondary response of T cells to inflammation and instead suggest that clonally expanded alphabeta T cells are implicated in mediating a component of the valvular injury responsible for CAS. PMID- 17404319 TI - Potential role for IL-7 in Fas-mediated T cell apoptosis during HIV infection. AB - IL-7 promotes survival of resting T lymphocytes and induces T cell proliferation in lymphopenic conditions. As elevated IL-7 levels occur in HIV-infected individuals in addition to high Fas expression on T cells and increased sensitivity to Fas-induced apoptosis, we analyzed whether IL-7 has a regulatory role in Fas-mediated T cell apoptosis. We show that IL-7 up-regulates Fas expression on naive and memory T cells through a mechanism that involves translocation of Fas molecules from intracellular compartments to the cell membrane. IL-7 induced the association of Fas with the cytoskeletal component ezrin and a polarized Fas expression on the cell surface. The potential role of IL-7 in Fas up-regulation in vivo was verified in IL-7-treated macaques and in HIV-infected or chemotherapy treated patients by the correlation between serum IL 7 levels and Fas expression on T cells. IL-7 treatment primed T cells for Fas induced apoptosis in vitro and serum IL-7 levels correlated with the sensitivity of T cells to Fas-induced apoptosis in HIV-infected individuals. Our data suggest an important role for IL-7 in Fas-mediated regulation of T cell homeostasis. Elevated IL-7 levels associated with lymphopenic conditions, including HIV infection, might participate in the increased sensitivity of T cells for activation-induced apoptosis. PMID- 17404320 TI - Some plasmin-induced antibodies bind to cardiolipin, display lupus anticoagulant activity and induce fetal loss in mice. AB - The combined presence of anti-phospholipid Ab (aPL), thrombosis, and/or fetal loss is recognized as the antiphospholipid syndrome (APS). aPL include anti cardiolipin Ab (aCL) and/or lupus anticoagulants (LAC, detected as Ig that prolong certain in vitro phospholipid (PL)-restricted blood clotting tests); both aCL and LAC are the diagnostic Ab for APS. Studies show that aPL represent a heterogeneous group of Ab, which recognize various PL, PL-binding plasma proteins, and/or PL-protein complexes. Recently, we found that five of seven patient-derived IgG monoclonal aCL react with thrombin, activated protein C, and plasmin. All three proteins are trypsin-like serine proteases (SP), and are highly homologous in their catalytic domains. Importantly, among these SP autoantigens, the reactive aCL bind to plasmin with the highest affinity, suggesting that plasmin may serve as a major driving autoantigen for some aCL in approximately 30% of APS patients who are positive for IgG anti-plasmin Ab. To test this hypothesis, we immunized BALB/c mice with human plasmin and analyzed immune sera for aCL activity and reactivity with relevant SP. We found that some immune sera displayed aCL activity and/or bound to test SP. Subsequently, eight mAb were obtained and studied. The results revealed that one mAb displayed the aCL and the LAC activities and induced fetal loss when injected into pregnant mice. Immunohistological analyses of placentas revealed extensive deposits of activated C3 components. Combined, these data demonstrate that plasmin may serve as a driving Ag for some pathogenic aPL. PMID- 17404321 TI - Overexpression of activation-induced cytidine deaminase in B cells is associated with production of highly pathogenic autoantibodies. AB - Defective receptor editing or defective B cell checkpoints have been associated with increased frequency of multireactive autoantibodies in autoimmune disease. However, Ig somatic hypermutation and/or class switch recombination may be mechanisms enabling the development of pathogenic multireactive autoantibodies. In this study, we report that, in the BXD2 mouse model of autoimmune disease, elevated expression of activation-induced cytidine deaminase (AID) in recirculating follicular CD86(+) subsets of B cells and increased germinal center B cell activity are associated with the production of pathogenic multireactive autoantibodies. CD4 T cells from BXD2 mice that expressed increased levels of CD28 and an increased proliferative response to anti-CD3 and anti-CD28 stimulation are required for this process. Inhibition of the CD28-CD86 interaction in BXD2 mice with AdCTLA4-Ig resulted in normalization of AID in the B cells and suppression of IgG autoantibodies. This treatment also prevented the development of germinal center autoantibody-producing B cells, suggesting that an optimal microenvironment enabling AID function is important for the formation of pathogenic autoantibodies. Taken together, our data indicate that AID expression in B cells is a promising therapeutic target for the treatment of autoimmune diseases and that suppression of this gene may be a molecular target of CTLA4-Ig therapy. PMID- 17404322 TI - Inhibition of transmethylation down-regulates CD4 T cell activation and curtails development of autoimmunity in a model system. AB - Transmethylation affects several cellular events, including T cell activation, and blockade of this pathway may curtail inflammatory/autoimmune responses. Here, we demonstrate that transmethylation inhibition by a novel reversible S-adenosyl l-homocysteine hydrolase inhibitor leads to immunosuppression by reducing phosphorylation of several key proteins involved in TCR signaling, including Akt, Erk1/2, and NF-kappaB. Remarkably, this effect was largely restricted to CD4 T cells and correlated with reduced arginine methylation of Vav1, an essential guanine nucleotide exchange factor in T cell stimulation. Treatment with the transmethylation inhibitor averted, and even ameliorated, the CD4-mediated autoimmune disease, experimental autoimmune encephalomyelitis. The data suggest that transmethylation is required for CD4 T cell activation, and its inhibition may be a novel approach in the treatment of multiple sclerosis, and other CD4 mediated autoimmune diseases. PMID- 17404323 TI - Airway exposure levels of lipopolysaccharide determine type 1 versus type 2 experimental asthma. AB - Allergic asthma is characterized by airway inflammation initiated by adaptive immune responses to aeroallergens. Recent data suggest that severe asthma may be a different form of asthma rather than an increase in asthma symptoms and that innate immune responses to LPS can modulate adaptive immune responses to allergens. In this study, we evaluated the hypothesis that airway exposure to different doses of LPS induces different form of asthma. Our study showed that neutrophilic inflammation and IFN-gamma expression were higher in induced sputum from severe asthma patients than from mild to moderate asthmatics. Animal experiments indicated that allergen sensitization with low-dose LPS (0.1 microg) induced type 2 asthma phenotypes, i.e., airway hyperresponsiveness, eosinophilic inflammation, and allergen-specific IgE up-regulation. In contrast, allergen sensitization with high-dose LPS (10 microg) induced asthma phenotypes, i.e., airway hyperresponsiveness and noneosinophilic inflammation that were not developed in IFN-gamma-deficient mice, but unaffected in the absence of IL-4. During the allergen sensitization period, TNF-alpha expression was found to be enhanced by both low- and high-dose LPS, whereas IL-12 expression was only enhanced by high-dose LPS. Interestingly, the asthma phenotypes induced by low dose LPS, but not by high-dose LPS, were completely inhibited in TNF-alpha receptor-deficient mice, whereas the asthma phenotypes induced by high-dose LPS were abolished in the homozygous null mutation of the STAT4 gene. These findings suggest that airway exposure levels of LPS induces different forms of asthma that are type 1 and type 2 asthma phenotypes by high and low LPS levels, respectively. PMID- 17404324 TI - Circulating immune complexes (IC) and IC-induced levels of GM-CSF are increased in sudanese patients with acute visceral Leishmania donovani infection undergoing sodium stibogluconate treatment: implications for disease pathogenesis. AB - Infection with Leishmania donovani is associated with IL-10 as well as with GM CSF. Immune complexes (IC) exert important functions by stimulation of monocytes/macrophage-mediated production of pro- and anti-inflammatory cytokines in rheumatic diseases. In this investigation, we have explored IC-induced cytokine production during Leishmania infection. Sera from 43 patients with visceral leishmaniasis (VL), 17 patients with post-kala-azar dermal leishmaniasis, and 20 healthy Sudanese controls were precipitated with polyethylene glycol (PEG). The PEG precipitates were added to serum-free PBMC for 20 h,whereupon supernatant levels of IL-1beta, IL-6, IL-10, IL-1 receptor antagonist protein, TNF-alpha, TNF receptor p75, and GM-CSF were investigated using ELISA. Circulating levels of C1q-binding IC were also measured in the serum samples. PEG precipitates from Leishmania-infected patients induced significantly higher levels of GM-CSF (p = 0.0037) and IL-10 (p < 0.0001), as well as of IL-6 (p < 0.0001) and IL-1 receptor antagonist (p = 0.0238) as compared with PEG precipitates from controls. Patients with acute VL as well as VL patients receiving sodium stibogluconate treatment displayed significantly increased levels of PEG precipitate-induced GM-CSF. The induction of GM-CSF by circulating IC was especially prominent in acute VL patients receiving sodium stibogluconate treatment; ANOVA revealed significant interaction between disease activity and treatment for PEG precipitate-induced levels of GM-CSF (disease activity, p = 0.0006; treatment, p = 0.0005; interaction, p = 0.0046). Parallel associations were determined for C1q-binding immune complexes, but not for any cytokine other than GM-CSF. The importance of IC-induced GM-CSF in leishmaniasis warrants further study. PMID- 17404325 TI - Amelioration of experimental autoimmune myasthenia gravis in rats by neonatal FcR blockade. AB - The neonatal FcR (FcRn) plays a critical role in IgG homeostasis by protecting it from a lysosomal degradation pathway. It has been shown that IgG has an abnormally short half-life in FcRn-deficient mice and that FcRn blockade significantly increases the catabolism of serum IgG in mice. Therefore, reduction of serum IgG half-life may have therapeutic benefits in Ab-mediated autoimmune diseases. We have studied the therapeutic effects of an anti-rat FcRn mAb, 1G3, in two rat models of myasthenia gravis, a prototypical Ab-mediated autoimmune disease. Passive experimental autoimmune myasthenia gravis was induced by administration of an anti-acetylcholine receptor (AChR) mAb, and it was shown that treatment with 1G3 resulted in dose-dependent amelioration of the disease symptoms. In addition, the concentration of pathogenic Ab in the serum was reduced significantly. The effect of 1G3 was also studied in an active model of experimental autoimmune myasthenia gravis in which rats were immunized with AChR. Treatment with 1G3 significantly reduced the severity of the disease symptoms as well as the levels of total IgG and anti-AChR IgG relative to untreated animals. These data suggest that FcRn blockade may be an effective way to treat Ab mediated autoimmune diseases. PMID- 17404326 TI - Inbreeding and genetic structure in the endangered Sorraia horse breed: implications for its conservation and management. AB - The Sorraia horse is a closed breed with reduced effective population size and considered in critical maintained risk status. The breed exists in 2 main breeding populations, one in Portugal and one in Germany, with a smaller population size. A set of 22 microsatellite loci was used to examine genetic diversity and structure of the Sorraia horse breed and to compare individual inbreeding coefficient F, estimated from pedigree data, with individual heterozygosity and mean d(2). The Sorraia horse shows lower levels of microsatellite diversity when compared with other horse breeds. Due to management strategies, there are clear differences in the genetic structure of the 2 main Sorraia horse populations. Individual heterozygosity was shown to be a good estimator, used together with or as an alternative to inbreeding coefficient, in predicting fitness and evaluating the inbreeding level of the Sorraia horse. The information gathered in this study, combined with information available from previous studies, offers an important and wide information base for the future development of an effective breeding management of the Sorraia horse in order to preserve this endangered breed. PMID- 17404327 TI - Establishment of an albino strain of the bitterling Tanakia signifer (Pisces, Cyprinidae). AB - An albino strain of the bitterling Tanakia signifer (Pisces, Cyprinidae, Acheilognathinae) was established through interbreeding using an albino male selected from among the offspring produced from the spawning of 18 wild-caught individuals. This is the first report of albinism in this species. Progeny tests demonstrated that inheritance of the trait follows the expected pattern for a simple autosomal recessive. A stock of 200+ individuals is currently being held at the Kannonzaki Nature History Museum, Japan. PMID- 17404328 TI - Variation and heritability in immune gene expression by diseased honeybees. AB - Social insects are frequent targets for pathogens and have consequently evolved diverse ways to minimize disease impacts, one of which is the innate immune response. Here, a 4-generation mating scheme was carried out to assess heritability and variation in a honeybee (Apis mellifera) immune trait, the production of the key antimicrobial peptide abaecin. Larval offspring from controlled crosses (n = 576 offspring from 36 singly inseminated queens) as well as offspring of field colonies (896 individuals in 53 colonies) were challenged individually with a widespread bee pathogen, the gram-positive bacterium Paenibacillus larvae. After bacterial challenge, transcript levels for the gene encoding abaecin were quantified and then compared using known pedigrees and colony environments. Considerable variation among highly related siblings (r = 0.75) indicates that subtle allelic differences in immune pathway genes can have large effects on transcriptional profiles. Abaecin levels were moderately heritable (h(2) = approximately 0.3-0.4), reflecting high amounts of standing genetic variation, and suggesting that this and other immune traits are amenable to selective programs aimed at improving honeybee health. The results help efforts to determine the relative effectiveness of social versus individual defenses by social insects toward their pathogens. PMID- 17404329 TI - Medical training: chaos and unemployment. PMID- 17404330 TI - The solution to chaos is not chaos. PMID- 17404331 TI - The treatment of enteric fever. PMID- 17404332 TI - Tempus fugit: competency assessment in Modernizing Medical Careers. PMID- 17404333 TI - Reed-Elsevier, academics and the arms trade. PMID- 17404334 TI - The value of an interview. PMID- 17404335 TI - Osborne and Craft duologue: relative values. PMID- 17404336 TI - Placebo management of psychogenic disease. PMID- 17404337 TI - Paying for cancer care - a new dilemma. PMID- 17404338 TI - A lesson learnt: the rise and fall of Lariam and Halfan. PMID- 17404339 TI - A cradle of reform. PMID- 17404340 TI - Bufale spotting, part two: assessing systematic reviews. PMID- 17404341 TI - Children with developmental coordination disorders. PMID- 17404342 TI - Reports of clinical trials should begin and end with up-to-date systematic reviews of other relevant evidence: a status report. AB - OBJECTIVE: Scientific and ethical justification for new clinical trials requires them to have been designed in the light of scientifically defensible assessments of relevant previous research. Reliable interpretation of the results of new clinical trials entails setting them in the context of updates of the reviews upon which they were deemed scientifically and ethically justifiable. We have shown previously that most reports of randomized trials published in five general medical journals in May 1997 and in May 2001 failed to set their results in the context of the findings from similar research. In the current study, we assess whether there had been progress in this respect in 2005 and also investigate the extent to which reports begin by referring to systematic reviews providing the justification for the new research reported. DESIGN: Assessment of the Introduction and Discussion sections in all reports of randomized trials published during May 2005 in five general medical journals. SETTING: Reports of randomized trials in five general medical journals. PARTICIPANTS: Annals of Internal Medicine, BMJ, JAMA, Lancet and New England Journal of Medicine. INTERVENTIONS: None. MAIN OUTCOME MEASURES: The inclusion or mention of one or more systematic reviews in the Introduction or Discussion section of each report assessed. RESULTS: We found 18 reports of randomized trials. The Introduction sections referred to systematic reviews in five (27%) of these reports. None of the discussion sections of the 15 reports of trials that were not the first published trials to address the question studied placed the results of the new trial in the context of an updated systematic review of other research. Although reference was made to relevant systematic reviews in five of these 15 reports, there was no integration - quantitative or qualitative - of the results of the new trials in an update of these reviews. In the remaining ten reports there was no evidence that any systematic attempt had been made to set the new results in the context of previous trials. CONCLUSIONS: There is no evidence of progress between 1997 and 2005 in the proportion of reports of trials published in general medical journals which discussed new results within the context of up-to-date systematic reviews of relevant evidence from other controlled trials. Although the proportion of trials referring to systematic reviews in Discussion sections has increased, the majority of reports continued to fail even to do this. Similarly, most researchers appear not to have considered a systematic review when designing their trial. Researchers and journal editors do a disservice to the interests of the public and others involved in healthcare decision-making by acquiescing in this situation. PMID- 17404343 TI - Sacral metastasis as a presentation of colonic adenocarcinoma. PMID- 17404344 TI - An early Medical Research Council controlled trial of vitamins for preventing infection. PMID- 17404345 TI - De Motu Cordis: the Lumleian Lecture of 1616: an imagined playlet concerning the discovery of the circulation of the blood by William Harvey. PMID- 17404346 TI - Summaries for patients. Exenatide therapy for type 2 diabetes. PMID- 17404347 TI - Summaries for patients. Screening mammography in women age 40 to 49 years. PMID- 17404348 TI - In the clinic. Allergic rhinitis. PMID- 17404349 TI - The effect of adding exenatide to a thiazolidinedione in suboptimally controlled type 2 diabetes: a randomized trial. AB - BACKGROUND: Exenatide therapy is effective in combination with metformin or sulfonylureas for treating type 2 diabetes. Thiazolidinediones (TZDs) also are commonly used, but the efficacy of exenatide with a TZD has not been reported. OBJECTIVE: To compare the effects of exenatide versus placebo on glycemic control. DESIGN: Placebo run-in, randomized, double-blind, placebo-controlled trial conducted from May 2004 to August 2005. SETTING: 49 sites in Canada, Spain, and the United States. PATIENTS: 233 (exenatide group, n = 121; placebo group, n = 112) patients with type 2 diabetes that was suboptimally controlled with TZD treatment (with or without metformin). Mean (+/-SE) baseline glycated hemoglobin A1c level was 7.9% +/- 0.1%. INTERVENTIONS: Subcutaneous abdominal injections of 10 microg of exenatide or placebo twice daily, added to a TZD (with or without metformin) for 16 weeks. MEASUREMENTS: The primary outcome was change from baseline in hemoglobin A1c level. Other outcomes were fasting serum glucose level, body weight, self-monitored blood glucose level, and any adverse events. RESULTS: Exenatide treatment reduced hemoglobin A(1c) level (mean difference, 0.98% [95% CI, -1.21% to -0.74%]), serum fasting glucose level (mean difference, 1.69 mmol/L [-30.5 mg/dL] [CI, -2.22 to -1.17 mmol/L {-40.0 to -21.1 mg/dL}]), and body weight (mean difference, -1.51 kg [CI, -2.15 to -0.88 kg]). Sixteen percent of patients in the exenatide group and 2% of patients in the placebo group discontinued treatment because of adverse events. In the exenatide group, 40% (n = 48) of patients experienced nausea (mostly mild [n = 21] or moderate [n = 19]), 13% experienced vomiting, and 11% experienced hypoglycemia. In the placebo group, 15% of patients experienced nausea, 1% experienced vomiting, and 7% experienced hypoglycemia. LIMITATIONS: Combinations with TZDs and sulfonylureas were not tested. Trial duration was relatively short. Only 71% and 86% of patients in the exenatide and placebo groups, respectively, completed the study. CONCLUSIONS: Exenatide therapy improved glycemic control, reduced body weight, and caused gastrointestinal symptoms more than placebo in patients with type 2 diabetes that was suboptimally controlled with TZD therapy. ClinicalTrials.gov registration number: NCT00099320. For more information on exenatide click here. PMID- 17404350 TI - Prevalence of neutropenia in the U.S. population: age, sex, smoking status, and ethnic differences. AB - BACKGROUND: Benign reductions in neutrophil counts may be more common at certain ages and in certain ethnic groups and may be affected by sex and smoking status. OBJECTIVE: To determine differences in neutrophil counts in the U.S. population according to ethnicity, age, sex, and smoking status. DESIGN: Population-based, cross-sectional study. SETTING: Various locations in the United States. PARTICIPANTS: 25,222 participants in the 1999 to 2004 National Health and Nutrition Examination Survey who were 1 year of age or older. MEASUREMENTS: Complete blood counts and comparison of means and the proportion of participants with neutropenia. RESULTS: Relative to white participants, black participants had lower leukocyte counts (mean difference, 0.89 x 10(9) cells/L; P < 0.001), lower neutrophil counts (0.83 x 10(9) cells/L; P < 0.001), and similar lymphocyte counts (0.022 x 10(9) cells/L; P = 0.36), whereas Mexican-American participants had slightly higher mean leukocyte counts (0.16 x 10(9) cells/L; P = 0.014), higher neutrophil counts (0.11 x 10(9) cells/L; P = 0.026), and higher lymphocyte counts (0.095 x 10(9) cells/L; P < 0.001). The prevalence of neutropenia (neutrophil count <1.5 x 10(9) cells/L) was 4.5% among black participants, 0.79% among white participants, and 0.38% among Mexican-American participants. The prevalence of neutropenia was higher among males and children younger than 5 years of age. Neutrophil counts less than 1.0 x 10(9) cells/L were observed in fewer than 1% of the overall sample (0.57% in black participants, 0.11% in white participants, and 0.08% in Mexican-American participants). Smoking was associated with higher leukocyte and neutrophil counts but had a smaller effect among black and Mexican-American participants than among white participants. LIMITATION: Because estimates are based on single measures, fluctuations over time could not be determined. CONCLUSIONS: In the United States, neutrophil counts are lower in black persons than in white persons and neutropenia is more prevalent in black persons. Neutrophil counts are slightly higher in Mexican-American persons than in white persons, and neutropenia is uncommon in both groups. The clinical implications of these findings are unclear, but they suggest that when determining the need for a diagnostic evaluation for neutropenia, clinicians should consider the patient's age, sex, ethnicity, and smoking status. PMID- 17404351 TI - Geography matters: relationships among urban residential segregation, dialysis facilities, and patient outcomes. AB - BACKGROUND: End-stage renal disease disproportionately affects black Americans. However, the impact of residential segregation by race-a prominent feature of many U.S. cities--on outcomes of patients receiving dialysis and on facility performance has not been evaluated. OBJECTIVE: To examine the relationship among racial composition of ZIP codes in metropolitan areas, outcomes of patients receiving dialysis, and characteristics of dialysis facilities. DESIGN: Retrospective cohort study of patients receiving dialysis and cross-sectional study of dialysis facilities. SETTING: U.S. metropolitan ZIP codes with differing percentages of black residents. PATIENTS: Black and non-Hispanic white patients who initiated long-term dialysis between 1 January 1995 and 31 December 2002 (n = 399,424) and dialysis facilities in operation in December 2004 (n = 3244). MEASUREMENTS: Mortality and time to transplantation among patients receiving dialysis, and performance of dialysis facilities on the basis of quality indicators (anemia management, dialysis adequacy, and facility-level mortality rates). RESULTS: Most black patients (50.3%) but few white patients (5%) lived in the 3% (n = 769) of ZIP codes in which most residents were black. In analyses adjusted for patient and ZIP code characteristics, mortality rates were higher among white patients but not among black patients living in areas with a higher percentage of black residents (adjusted hazard ratio for ZIP codes with > or =75% black residents vs. <10% black residents, 1.14 [95% CI, 1.07 to 1.21] for white patients and 1.02 [CI, 0.99 to 1.06] for black patients). Time to transplantation was longer among both black and white patients (adjusted hazard ratio for ZIP codes with > or =75% black residents vs. <10% black residents, 0.84 [CI, 0.78 to 0.92] and 0.63 [CI, 0.57 to 0.71] for black patients and white patients, respectively). Dialysis facilities located in areas with a higher percentage of black residents were more likely to have higher-than-expected mortality rates and were less likely to meet performance targets. LIMITATIONS: Patient-level analyses were restricted to black and non-Hispanic white patients. Patient-level and facility-level analyses focused only on the percentage of black residents in each ZIP code. CONCLUSIONS: The racial composition of urban residential areas is associated with time to transplantation and dialysis facility performance on standard quality measures. Closer scrutiny of care provided to patients receiving dialysis who live in predominantly black residential areas and to dialysis facilities operating in these areas may be warranted. PMID- 17404352 TI - Systematic review: the long-term effects of false-positive mammograms. AB - BACKGROUND: Although abnormal screening mammograms deleteriously affect the psychological well-being of women during the time immediately surrounding the tests, their long-term effects are poorly understood. PURPOSE: To characterize the long-term effects of false-positive screening mammograms on the behavior and well-being of women 40 years of age or older. DATA SOURCES: English-language studies from the MEDLINE, Web of Science, EMBASE, CINAHL, PsycINFO, and ERIC databases through August 2006. STUDY SELECTION: Studies were identified that examined the effects of false-positive results of routine screening mammography on women's behavior, well-being, or beliefs. DATA EXTRACTION: Two investigators independently coded study characteristics, quality, and effect sizes. DATA SYNTHESIS: 23 eligible studies (n = 313,967) were identified. A random-effects meta-analysis showed that U.S. women who received false-positive results on screening mammography were more likely to return for routine screening than those who received normal results (risk ratio, 1.07 [95% CI, 1.02 to 1.12]). The effect was not statistically significant among European women (risk ratio, 0.97 [CI, 0.93 to 1.01]), and Canadian women were less likely to return for routine screening because of false-positive results (risk ratio, 0.63 [CI, 0.50 to 0.80]). Women who received false-positive results conducted more frequent breast self-examinations and had higher, but not apparently pathologically elevated, levels of distress and anxiety and thought more about breast cancer than did those with normal results. LIMITATIONS: Correlational study designs, a small number of studies, a lack of clinical validation for many measures, and possible heterogeneity. CONCLUSIONS: Some women with false-positive results on mammography may have differences in whether they return for mammography, occurrence of breast self-examinations, and levels of anxiety compared with women with normal results. Future research should examine how false-positive results on mammography affect other outcomes, such as trust and health care use. PMID- 17404353 TI - Screening mammography for women 40 to 49 years of age: a clinical practice guideline from the American College of Physicians. AB - Breast cancer is one of the most common causes of death for women in their 40s in the United States. Individualized risk assessment plays an important role when making decisions about screening mammography, especially for women 49 years of age or younger. The purpose of this guideline is to present the available evidence for screening mammography in women 40 to 49 years of age and to increase clinicians' understanding of the benefits and risks of screening mammography. PMID- 17404354 TI - Screening mammography in women 40 to 49 years of age: a systematic review for the American College of Physicians. AB - BACKGROUND: The risks and benefits of mammography screening among women 40 to 49 years of age remain an important issue for clinical practice. PURPOSE: To evaluate the evidence about the risks and benefits of mammography screening for women 40 to 49 years of age. DATA SOURCES: English-language publications in MEDLINE (1966-2005), Pre-MEDLINE, and the Cochrane Central Register of Controlled Trials and references of selected studies through May 2005. STUDY SELECTION: Previous systematic reviews; randomized, controlled trials; and observational studies. DATA EXTRACTION: Two independent reviewers. DATA SYNTHESIS: In addition to publications from the original mammography trials, 117 studies were included in the review. Meta-analyses of randomized, controlled trials demonstrate a 7% to 23% reduction in breast cancer mortality rates with screening mammography in women 40 to 49 years of age. Screening mammography is associated with an increased risk for mastectomy but a decreased risk for adjuvant chemotherapy and hormone therapy. The risk for death due to breast cancer from the radiation exposure involved in mammography screening is small and is outweighed by a reduction in breast cancer mortality rates from early detection. Rates of false positive results are high (20% to 56% after 10 mammograms), but false-positive results have little effect on psychological health or subsequent mammography adherence. Although many women report pain at the time of the mammography, few see pain as a deterrent to future screening. Evidence about the effect of negative screening mammography on psychological well-being or the subsequent clinical presentation of breast cancer is insufficient. LIMITATIONS: Few randomized, controlled trials assessed the risks of screening, and the literature search was completed in 2005. CONCLUSIONS: Although few women 50 years of age or older have risks from mammography that outweigh the benefits, the evidence suggests that more women 40 to 49 years of age have such risks. PMID- 17404355 TI - Exenatide in combination therapy: small study, big market, and many unanswered questions. PMID- 17404356 TI - Breast cancer screening for women in their 40s: moving from controversy about data to helping individual women. PMID- 17404357 TI - Trials that matter: two faces of progress in the treatment of age-related macular degeneration. PMID- 17404358 TI - Kayla. PMID- 17404359 TI - Ethically increasing the supply of transplantable organs. PMID- 17404360 TI - Ethically increasing the supply of transplantable organs. PMID- 17404361 TI - Does pay-for-performance improve the quality of health care? PMID- 17404362 TI - Discordance between sexual behavior and self-reported sexual identity. PMID- 17404363 TI - Discordance between sexual behavior and self-reported sexual identity. PMID- 17404364 TI - Hypocalcemia as a cause of reversible cardiomyopathy with ventricular tachycardia. PMID- 17404365 TI - Initial hormonal management of androgen-sensitive metastatic, recurrent, or progressive prostate cancer: 2006 update of an American Society of Clinical Oncology practice guideline. AB - PURPOSE: To update the 2004 American Society of Clinical Oncology (ASCO) guideline on initial hormonal management of androgen-sensitive, metastatic, recurrent, or progressive prostate cancer (PCa). METHODS: The writing committee based its recommendations on an updated systematic literature review. Recommendations were approved by the Expert Panel, the ASCO Health Services Committee, and the ASCO Board of Directors. RESULTS: Seven randomized controlled trials (four new), one systematic review, one meta-analysis (new), one Markov model, and one delta-method 95% CI procedure for active controlled trials (new) informed the guideline update. RECOMMENDATIONS: Bilateral orchiectomy or luteinizing hormone-releasing hormone agonists are recommended initial androgen deprivation treatments (ADTs). Nonsteroidal antiandrogen monotherapy merits discussion as an alternative; steroidal antiandrogen monotherapy should not be offered. Combined androgen blockade should be considered. In metastatic or progressive PCa, immediate versus symptom-onset institution of ADT results in a moderate decrease (17%) in relative risk (RR) for PCa-specific mortality, a moderate increase (15%) in RR for non-PCa-specific mortality, and no overall survival advantage. Therefore, the Panel cannot make a strong recommendation for early ADT initiation. Prostate-specific antigen (PSA) kinetics and other metrics allow identification of populations at high risk for PCa-specific and overall mortality. Further studies must be completed to assess whether patients with adverse prognostic factors gain a survival advantage from immediate ADT. For patients electing to wait until symptoms for ADT, regular monitoring visits are indicated. For patients with recurrence, clinical trials should be considered if available. Currently, data are insufficient to support use of intermittent androgen blockade outside clinical trials PMID- 17404366 TI - Adjuvant chemotherapy for high-risk operable breast cancer. PMID- 17404367 TI - Breast cancer mortality trends in the United States according to estrogen receptor status and age at diagnosis. AB - PURPOSE: Since 1990, overall breast cancer mortality rates in the United States decreased 24%. This decline has been attributed to mammography screening and adjuvant systemic therapy. However, the efficacy of these modalities may depend on estrogen receptor (ER) expression and age. We therefore examined breast cancer mortality trends in the United States according to ER status and age. METHODS: Using the Surveillance, Epidemiology, and End Results (SEER) program (1990-2003), we calculated trends in incidence-based mortality (IBM), annual hazard rates for breast cancer deaths after diagnosis, and relative hazard rates for women with ER positive and ER-negative tumors. Relative hazard rates were assessed with Cox proportional hazards models, adjusted for stage and grade, and stratified by age at diagnosis. RESULTS: During the study period, IBM and annual hazard rates for breast cancer deaths decreased among women with ER-positive and ER-negative tumors, although declines were greater for those with ER-positive tumors. Among women younger than 70 years, relative hazard rates declined 38% for those with ER positive tumors versus 19% for those with ER-negative tumors. Among women 70 years or older, relative hazard rates declined 14% for those with ER-positive tumors versus no significant decline for those with ER-negative tumors. CONCLUSION: In the United States, breast cancer mortality rates have declined among women with ER-positive and ER-negative tumors, with greater declines among younger women and those with ER-positive tumors. Although mortality in all groups remains unacceptably high, additional emphasis should be placed on improving outcomes of breast cancer patients older than 70 years and those of all ages with ER-negative tumors. PMID- 17404368 TI - Intensive dose-dense compared with high-dose adjuvant chemotherapy for high-risk operable breast cancer: Southwest Oncology Group/Intergroup study 9623. AB - PURPOSE: Southwest Oncology Group (SWOG)/Intergroup study 9623 was undertaken to compare treatment with an anthracycline-based adjuvant chemotherapy regimen followed by high-dose chemotherapy (HDC) with autologous hematopoietic progenitor cell support (AHPCS) with a modern dose-dense dose-escalated (nonstandard) regimen including both an anthracycline and a taxane. PATIENTS AND METHODS: Participants in this phase III randomized study had operable breast cancer involving four or more axillary lymph nodes and had completed mastectomy or breast-conserving surgery. Patients were randomly assigned to receive four cycles of doxorubicin and cyclophosphamide followed by HDC with AHPCS or to receive sequential dose-dense and dose-escalated chemotherapy with doxorubicin, paclitaxel, and cyclophosphamide. The primary end point of this study was disease free survival (DFS). RESULTS: Among 536 eligible patients, there was no significant difference between the two arms for DFS or overall survival (OS). Estimated five-year DFS was 80% (95% CI, 76% to 85%) for dose-dense therapy and 75% (95% CI, 69% to 80%) for transplantation. Estimated 5-year OS was 88% (95% CI, 84% to 92%) for dose-dense therapy and 84% (95% CI, 79% to 88%) for transplantation. CONCLUSION: There is no evidence that transplantation was superior to dose-dense dose-escalated therapy. Transplantation was associated with an increase in toxicity and a possibly inferior outcome, although the hazard ratios were not significantly different from 1. PMID- 17404369 TI - Induction chemotherapy followed by chemoradiotherapy compared with chemoradiotherapy alone for regionally advanced unresectable stage III Non-small cell lung cancer: Cancer and Leukemia Group B. AB - PURPOSE: Standard therapy for unresectable stage III non-small-cell lung cancer includes concomitant chemoradiotherapy. In Cancer and Leukemia Group B 39801, we evaluated whether induction chemotherapy before concurrent chemoradiotherapy would result in improved survival. PATIENTS AND METHODS: Between July 1998 and May 2002, 366 patients were randomly assigned to arm A, which involved immediate concurrent chemoradiotherapy with carboplatin area under the concentration-time curve (AUC) of 2 and paclitaxel 50 mg/m2 given weekly during 66 Gy of chest radiotherapy, or arm B, which involved two cycles of carboplatin AUC 6 and paclitaxel 200 mg/m2 administered every 21 days followed by identical chemoradiotherapy. The accrual goal was 360 patients. RESULTS: Thirty-four percent of patients were female, 66% were male, and the median age was 63 years. Grade 3 or 4 toxicities during induction chemotherapy on arm B consisted mainly of neutropenia (18% and 20%, respectively). During concurrent chemoradiotherapy, there was no difference in severity of in-field toxicities of esophagitis (grade 3 and 4 were, respectively, 30% and 2% for arm A v 28% and 8% for arm B) and dyspnea (grade 3 and 4 were, respectively, 11% and 3% for arm A v 15% and 4% for arm B). Survival differences were not statistically significant (P = .3), with a median survival on arm A of 12 months (95% CI, 10 to 16 months) versus 14 months (95% CI, 11 to 16 months) on arm B and a 2-year survival of 29% (95% CI, 22% to 35%) and 31% (95% CI, 25% to 38%). Age, weight loss before therapy, and performance status were statistically significant predictive factors. CONCLUSION: The addition of induction chemotherapy to concurrent chemoradiotherapy added toxicity and provided no survival benefit over concurrent chemoradiotherapy alone. The median survival achieved in each of the treatment groups is low, and the routine use of weekly carboplatin and paclitaxel with simultaneous radiotherapy should be re-examined. PMID- 17404370 TI - Gene expression profile in temporal lobe epilepsy. AB - Epilepsy is one of the most common neurological disorders. Temporal lobe epilepsy (TLE) represents the most frequent epilepsy syndrome in adult patients with resistance to pharmacological treatment. In TLE, the origin of seizure activity typically involves the hippocampal formation, which displays major neuropathological features, described with the term hippocampal sclerosis (HS). The expansion of neurosurgical epilepsy programs has offered the possibility of disposing of clinically well-characterized hippocampal tissue, so that the analysis of molecular mechanisms underlying the structural and functional reorganization occurring in the hippocampus and neighboring areas in TLE patients can be done on a large scale. The recent development of molecular biological technologies permits the analysis of changes in the expression of a large number of genes. This has opened new perspectives for epilepsy research. However, the hippocampal specimens obtained from patients with TLE most often represent an advanced stage of the pathology. For this reason, animal models that reproduce the clinical and histopathological features of TLE are helpful in detecting the early development of the pathological cascade leading to TLE with HS. An overview of recent data of gene expression profiles in human and experimental TLE is presented along with a discussion of the relevance of functional genomics, to develop new hypotheses and to detect likely candidate genes involved in epileptogenesis, as well as possible target molecules for new therapeutic approaches. PMID- 17404371 TI - The endocannabinoid system and neurogenesis in health and disease. AB - The endocannabinoid system exerts an important neuromodulatory function in different brain areas and is also known to be involved in the regulation of neural cell fate. Thus, CB(1) cannabinoid receptors are neuroprotective in different models of brain injury, and their expression is altered in various neurodegenerative diseases. Recent findings have demonstrated the presence of a functional endocannabinoid system in neural progenitor cells that participates in the regulation of cell proliferation and differentiation. In this Research Update, the authors address the experimental evidence regarding the regulatory role of cannabinoids in neurogenesis and analyze them in the context of those pathological disorders in which cannabinoid function and altered neuronal or glial generation is most relevant, for example, stroke and multiple sclerosis. PMID- 17404372 TI - Matching of pre- and postsynaptic specializations during synaptogenesis. AB - Formation of chemical synapses in the central nervous system is a highly regulated, multistep process that requires bidirectional communication across the synaptic cleft. Neurotransmitter receptors, scaffolding proteins, and signaling molecules need to be concentrated in the postsynaptic density, a specialized membrane microdomain apposed to the active zone of presynaptic terminals, where transmitter release occurs. This precise, synapse-specific matching implicates that sorting and targeting mechanisms exist for the molecular constituents of different types of synapses to ensure correct formation of neuronal circuits in the brain. There is considerable evidence from in vitro and in vivo studies that neurotransmitter signaling is not required for proper sorting during synapse formation, whereas active neurotransmission is essential for long-term synapse maintenance. Here, the authors review recent studies on the role of cell adhesion molecules in synaptogenesis and on possible mechanisms ensuring correct matching of pre- and postsynaptic sites. They discuss the role of neurotransmitter receptors and scaffolding proteins in these processes, focusing on fundamental differences between synapse formation during development and synapse maintenance and plasticity in adulthood. PMID- 17404373 TI - Endocannabinoids and synaptic function in the CNS. AB - Marijuana affects neural functions through the binding of its active component (Delta(9)-THC) to cannabinoid receptors in the CNS. Recent studies have elucidated that endogenous ligands for cannabinoid receptors, endocannabinoids, serve as retrograde messengers at central synapses. Endocannabinoids are produced on demand in activity-dependent manners and released from postsynaptic neurons. The released endocannabinoids travel backward across the synapse, activate presynaptic CB1 cannabinoid receptors, and modulate presynaptic functions. Retrograde endocannabinoid signaling is crucial for certain forms of short-term and long-term synaptic plasticity at excitatory or inhibitory synapses in many brain regions, and thereby contributes to various aspects of brain function including learning and memory. Molecular identities of the CB1 receptor and enzymes involved in production and degradation of endocannabinoids have been elucidated. Anatomical studies have demonstrated unique distributions of these molecules around synapses, which provide morphological bases for the roles of endocannabinoids as retrograde messengers. CB1-knockout mice exhibit various behavioral abnormalities and multiple defects in synaptic plasticity, supporting the notion that endocannabinoid signaling is involved in various aspects of neural function. In this review article, the authors describe molecular mechanisms of the endocannabinoid-mediated synaptic modulation and its possible physiological significance. PMID- 17404374 TI - Rethinking cortical organization: moving away from discrete areas arranged in hierarchies. AB - One way to understand the topography of the cerebral cortex is that "like attracts like." The cortex is organized to maximize nearest neighbor similarity. This principle can explain the separation of the cortex into discrete areas that emphasize different information domains. It can also explain the maps that form within cortical areas. However, because the cortex is two-dimensional, when a parameter space of much higher dimensionality is reduced onto the cortical sheet while optimizing nearest neighbor relationships, the result may lack an obvious global ordering into separate areas. Instead, the topography may consist of partial gradients, fractures, swirls, regions that resemble separate areas in some ways but not others, and in not a lack of topographic maps but an excess of maps overlaid on each other, no one of which seems to be entirely correct. Like a canvas in a gallery of modern art that no two observers interpret the same way, this lack of obvious ordering of high-dimensional spaces onto the cortex might then result in some scientific controversy over the true organization. In this review, the authors suggest that at least some sectors of the cortex do not have a simple global ordering and are better understood as a result of a reduction of a high-dimensional space onto the cortical sheet. The cortical motor system may be an example of this phenomenon. The authors discuss a model of the lateral motor cortex in which a reduction of many parameters onto a simulated cortical sheet results in a complex topographic pattern that matches the actual monkey motor cortex in surprising detail. Some of the ambiguities of topography and areal boundaries that have plagued the attempt to systematize the lateral motor cortex are explained by the model. PMID- 17404375 TI - The nucleus accumbens and Pavlovian reward learning. AB - The ability to form associations between predictive environmental events and rewarding outcomes is a fundamental aspect of learned behavior. This apparently simple ability likely requires complex neural processing evolved to identify, seek, and use natural rewards and redirect these activities based on updated sensory information. Emerging evidence from both animal and human research suggests that this type of processing is mediated in part by the nucleus accumbens (NAc) and a closely associated network of brain structures. The NAc is required for a number of reward-related behaviors and processes specific information about reward availability, value, and context. In addition, this structure is critical for the acquisition and expression of most Pavlovian stimulus-reward relationships, and cues that predict rewards produce robust changes in neural activity in the NAc. Although processing within the NAc may enable or promote Pavlovian reward learning in natural situations, it has also been implicated in aspects of human drug addiction, including the ability of drug paired cues to control behavior. This article provides a critical review of the existing animal and human literature concerning the role of the NAc in Pavlovian learning with nondrug rewards and considers some clinical ture concerning the role of the NAc in Pavlovian learning with nondrug implications of these findings. PMID- 17404376 TI - Neural systems for error monitoring: recent findings and theoretical perspectives. AB - Complex behavior requires a flexible system that maintains task performance in the context of specific goals, evaluating behavioral progress, adjusting behavior as needed, and adapting to changing contingencies. Generically referred to as performance monitoring, a key component concerns the identification and correction of differences between an intended and an executed response (i.e., an error). Brain mapping experiments have now identified the temporal and spatial components of a putative error-processing system in the large-scale networks of the human brain. Most of this work has focused on the medial frontal cortex and an associated electrophysiological component known as the error-related negativity (or error negativity). Although the precise role, or roles, of this region still remain unknown, investigations of error processing have identified a cluster of modules in the medial frontal cortex involved in monitoring/maintaining ongoing behavior and motivating task sets. Other regions include bilateral anterior insula/inferior operculum and lateral prefrontal cortex. Recent work has begun to uncover how individual differences might affect the modules recruited for a task, in addition to the identification of associations between pathological states and aberrant error signals, leading to insights about possible mechanisms of neuropsychiatric illness. PMID- 17404377 TI - Astroglial cells in development, regeneration, and repair. AB - Three main cellular components have been described in the CNS: neurons, astrocytes, and oligodendrocytes. In the past 10 years, lineage studies first based on retroviruses in the embryonic CNS and then by genetic fate mapping in both the prenatal and postnatal CNS have proposed that astroglial cells can be progenitors for neurons and oligodendrocytes. Hence, the population of astroglial cells is increasingly recognized as heterogeneous and diverse, encompassing cell types performing widely different roles in development and plasticity. Astroglial cells populating the neurogenic niches increase their proliferation after perinatal injury and in young mice can differentiate into neurons and oligodendrocytes that migrate to the cerebral cortex, replacing the cells that are lost. Although much remains to be learned about this process, it appears that the up-regulation of the Fibroblast growth factor receptor is critical for mediating the injury-induced increase in cell division and perhaps for the neuronal differentiation of astroglial cells. PMID- 17404378 TI - Tissue-specific expression of SORBITOL DEHYDROGENASE in apple fruit during early development. AB - Sorbitol, the primary photosynthate and translocated carbohydrate in apple (Malusxdomestica Borkh.), is converted to fructose by sorbitol dehydrogenase (SDH; EC 1.1.1.14) which is active in apple fruit throughout development. In the apple genome, nine SDH genes have been isolated and their sequences characterized, but their individual expression patterns during apple fruit set and development have not been determined. The objective of this work was to ascertain if SDH genes are differentially expressed and how their patterns of expression may relate to SDH activity in apple seed and cortex during early fruit development. Seed SDH activity was found to be much higher than cortex SDH activity per mg and g fresh weight (FW), and seed SDH activity contributed significantly to whole fruit SDH activity during weeks 2-5 after bloom. Five of the nine SDH genes present in the apple genome were expressed in apple fruit. Two SDH genes, SDH1 and SDH3, were expressed in both seed and cortex tissues. SDH2 expression was limited to cortex, while SDH6 and SDH9 were expressed in seed tissues only. SDH isomeric proteins of different pI values were detected in apple fruit. SDH isomers with pI values of 4.2, 4.8, 5.5, and 6.3 were found in seeds, and SDH isomers with pI values of 5.5, 6.3, 7.3, and 8.3 were found in cortex. The present work is the first to show that SDH is highly active in apple seed and that SDH genes are differentially expressed in seed and cortex during early development. PMID- 17404379 TI - 9-lipoxygenase metabolism is involved in the almond/Aspergillus carbonarius interaction. AB - Phyto-oxylipins are a group of biologically active molecules that play an important role in plant defence. Their production begins with the oxygenation of polyunsaturated fatty acids by lipoxygenases (LOX) to form 9- or 13 hydroperoxides that are substrates for several enzymes involved in the synthesis of final oxylipins, which can act as signal molecules and/or direct antimicrobials. In the present work, the response of the 9-LOX pathway in the almond/Aspergillus carbonarius (producer of ochratoxin A) interaction was studied. Both LOX gene expression and activity are up-regulated over the course of fungal infection in immature and mature almonds. The biochemical characterization of major LOX and hydroperoxide lyase (HPL) isoforms indicated that 9-LOX metabolism is specifically induced by A. carbonarius. Lipid peroxidation profiling showed that, in infected immature almonds, enzymatically produced 9-hydro(peroxy) fatty acids (HFAs) were the main HFAs and are further metabolized by HPL into C9-aldehydes. Both HPL gene expression and C9-aldehydes increased over the course of fungal infection. In mature almonds infected with A. carbonarius, levels of LOX expression and activity were lower than those found in immature seeds, and 9-HFA represented the minority of total HFA, which consisted of mostly 13- and non-enzymatically produced HFA. In these experimental conditions, no volatile aldehydes were recorded from these samples, even though HPL was up-regulated in infected mature almonds. The effects on the growth of A. carbonarius of the aldehydes produced by these enzymes were also tested in vitro. Results reported here led to the proposal that, in almond seed, the association of 9-LOX and HPL has an important role in seed defence mechanism against pathogen infection. PMID- 17404380 TI - Can dual chlorophyll fluorescence excitation be used to assess the variation in the content of UV-absorbing phenolic compounds in leaves of temperate tree species along a light gradient? AB - The present study assesses light-induced variations in phenolic compounds in leaves of saplings of two co-occurring temperate species (Acer platanoides L., and Fraxinus excelsior L.) along a light gradient using a new non-invasive optical method (Dualex). The Dualex-derived UV absorbance of leaf epidermis (the sum of the adaxial and abaxial faces, AUV) increased significantly with increasing light in both species. AUV values were correlated with absorbance of the leaf extract at 305 nm and 375 nm (A305 and A375) in both species with similar slopes for both species. However, a large difference in intercept was observed between the two species when A305 was regressed against AUV. Similarly, AUV values were well correlated with the amount of phenolics in the leaf extracts assessed by the Folin-Ciocalteu method, but slopes were significantly different for the two species. Thus, the UV-A epidermal transmittance, despite being a reliable indicator of the UV-screening capacity of the leaf epidermis, cannot be used for any quantitative estimate of UV-B screening capacity or of energetic requirement for leaf construction without a species-specific calibration. PMID- 17404381 TI - Control of scion vigour by kiwifruit rootstocks is correlated with spring root pressure phenology. AB - Root pressure was measured continuously over spring in eight clonal kiwifruit rootstocks selected from seven Actinidia species (A. chrysantha, A. deliciosa, A. eriantha, A. hemsleyana, A. kolomikta, A. macrosperma, A. polygama), using pressure transducers and miniature compression fittings. Rootstocks that promoted scion vigour developed root pressures up to 0.15 MPa before or during scion budburst, whereas those that reduced scion vigour developed root pressure up to 0.05 MPa only after scion shoot expansion. When several seasons were compared, the date of onset of root pressure and the magnitude of pressure achieved were consistent for each rootstock. Root pressure was first recorded between late July and early September in vigour-promoting rootstocks, while scion budburst and initial shoot growth were in late August and early September. Vigour-reducing rootstocks did not develop significant root pressure until October. The date of onset was similar for the grafted rootstock and ungrafted plant of the same clone, but was not clearly related to the timing of shoot growth by the ungrafted plant. In the grafted plants the leaf and xylem water potentials of the scion were more negative, midday turgor was 0.3-0.5 MPa lower, and wilting was sometimes observed in developing shoots growing on low-vigour rootstocks, indicating that water stress was contributing to reductions in growth. Leaf turgor was correlated with average root pressure but not pressure measured during the day, suggesting that root pressure was not supporting transpiration during peak flows and was, instead, indicative of higher root hydraulic conductance. The rapid temporal rise in root pressure observed each spring in the various rootstocks was not accompanied by changes in xylem sap solute potential, but when rootstock clones were compared those that developed higher root pressures had higher sap solute potentials. Xylem sap solute potential varied between rootstocks from -0.07 MPa to -0.15 MPa, while root pressures measured at the same time varied between 0.0 MPa and 0.09 MPa, suggesting that an osmotic mechanism could account for the observed root pressure. Differences in phenology between the rootstocks and scion appeared to account for the rootstock effects on shoot growth, and changes in root pressure provided a useful indication of seasonal changes in root hydraulic properties and solute transport behaviour. PMID- 17404382 TI - Expression and functional analysis of metal transporter genes in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens. AB - Zinc (Zn) hyperaccumulation is a constitutive property of Thlaspi caerulescens, whereas cadmium (Cd) hyperaccumulation varies greatly among different ecotypes. The molecular basis of this variation is unknown. Ecotypic differences in the sequences and expression of four representative ZIP family transporter genes were investigated. Genome analysis indicated the presence of at least two closely related copies of the TcIRT1 gene in both Ganges (high Cd accumulating) and Prayon (low Cd accumulating) ecotypes, with different copies being expressed in each, and, furthermore, the two genes potentially encode different length transcripts. The predominant transcript in Prayon was truncated, missing sequence coding for the putative metal-binding site and the five C-terminal transmembrane helices. The two ecotypes were grown hydroponically +/-Fe and Cd, and mRNA abundance determined for four ZIP genes. The four ZIP genes studied (TcIRT1, TcIRT2, TcZNT1, and TcZNT5) were expressed in roots only. TcIRT1 expression (full length in Ganges, TcIRT1-1G; truncated in Prayon, TcIRT1-2P) was enhanced by Fe deficiency or by exposure to Cd. TcIRT2 expression was induced by Fe deficiency, but was unaffected by Cd exposure. TcZNT5-G showed greater expression in Prayon compared with Ganges. The functions of TcIRT1 from Ganges and Prayon and the Arabidopsis homologue were analysed by heterologous expression in yeast. All three IRT1 genes were able to facilitate growth on low Fe concentrations. Cd sensitivity of yeast was conferred in the order AtIRT1>TcIRT1-1G>TcIRT1-2P (truncated). Cd uptake after 4 h was only detectable following complementation by AtIRT1. The results suggest that although TcIRT1-G may be involved in Cd hyperaccumulation in the Ganges ecotype of T. caerulescens, the transporter expressed in yeast does not have an enhanced ability to transport Cd compared with AtIRT1. Therefore, the unique Cd-accumulating ability of the T. caerulescens Ganges ecotype must be due to the levels of expression of the protein or to other factors such as interacting proteins. PMID- 17404383 TI - Both chloronemal and caulonemal cells expand by tip growth in the moss Physcomitrella patens. AB - Tip growth is a mode of cell expansion in which all growth is restricted to a small area that forms a tip in an elongating cell. In green plants, tip growth has been shown to occur in root hairs, pollen tubes, rhizoids, and caulonema. Each of these cell types has a longitudinally elongated shape, longitudinally oriented microtubules and actin microfilaments, and a characteristic cytoplasmic organization at the growing tip which is required for growth. Chloronema are elongated cylindrical shaped cells that form during the development of the moss protonema. Since there are no published reports on the precise mode of chloronema elongation and conflicting interpretations of its cytology, the mechanism of cell growth has remained unclear. To determine if chloronema elongate by tip or diffuse growth, time-lapse light microscopy was employed to follow the movement of fluorescent microspheres attached to the surface of growing cells. It is shown here that chloronemal cells elongate by a form of tip growth. However, the slower growth of chloronema compared with caulonema is probably the result of differences in cytological organization of the growing tip. PMID- 17404384 TI - Suppression subtractive hybridization identifies genes induced in response to UV B irradiation in apple skin: isolation of a putative UDP-glucose 4-epimerase. AB - Suppression subtractive hybridization (SSH) successfully identified 11 cDNAs in apple skin with highly induced expression as a result of ultraviolet (UV)-B irradiation. Apart from three putative flavonoid biosynthetic genes, chalcone synthase (CHS; A5C), flavanone-3-hydroxylase (F3H; B5F), and flavonol synthase (FLS; D1F), five clones (A1H, A10E, B11G, D5F, and D11H) were induced by low temperature (17 degrees C) as well, which is also known to induce anthocyanin accumulation in apple skin. Moreover, four clones (A1H, A10E, B11G, and D11H), showing higher expression levels in the skin, accumulated higher anthocyanin concentrations than their counterparts. Of the four clones, only A10E, a putative UDP-glucose 4-epimerase (UGE), was deemed to play an important role in anthocyanin accumulation in apple skin based on the facts that: (i) its transcription level was higher in the deep red cultivar, 'Jonathan', than in the pale red cultivar, 'Tsugaru'; and (ii) it could reversibly catalyse UDP-glucose to UDP-galactose, and the latter molecule is a major sugar donor for cyanidin glycoside in apple. Therefore, the full-length cDNA of A10E was isolated by rapid amplification of cDNA ends (RACE) and designated as MdUGE1. Further analysis demonstrated that UGE enzymatic activity was positively correlated with anthocyanin accumulation in apple skin. Thus, MdUGE1 isolated by SSH could play an important role in anthocyanin biosynthesis in apple skin in concert with other flavonoid biosynthetic genes. PMID- 17404385 TI - Opioid receptors mediate direct predictive fear learning: evidence from one-trial blocking. AB - Pavlovian fear learning depends on predictive error, so that fear learning occurs when the actual outcome of a conditioning trial exceeds the expected outcome. Previous research has shown that opioid receptors, including mu-opioid receptors in the ventrolateral quadrant of the midbrain periaqueductal gray (vlPAG), mediate such predictive fear learning. Four experiments reported here used a within-subject one-trial blocking design to study whether opioid receptors mediate a direct or indirect action of predictive error on Pavlovian association formation. In Stage I, rats were trained to fear conditioned stimulus (CS) A by pairing it with shock. In Stage II, CSA and CSB were co-presented once and co terminated with shock. Two novel stimuli, CSC and CSD, were also co-presented once and co-terminated with shock in Stage II. The results showed one-trial blocking of fear learning (Experiment 1) as well as one-trial unblocking of fear learning when Stage II training employed a higher intensity footshock than was used in Stage I (Experiment 2). Systemic administrations of the opioid receptor antagonist naloxone (Experiment 3) or intra-vlPAG administrations of the selective mu-opioid receptor antagonist CTAP (Experiment 4) prior to Stage II training prevented one-trial blocking. These results show that opioid receptors mediate the direct actions of predictive error on Pavlovian association formation. PMID- 17404386 TI - Isoform specificity of protein kinase Cs in synaptic plasticity. AB - Protein kinase Cs (PKCs) are implicated in many forms of synaptic plasticity. However, the specific isoform(s) of PKC that underlie(s) these events are often not known. We have used Aplysia as a model system in order to investigate the isoform specificity of PKC actions due to the presence of fewer isoforms and a large number of documented physiological roles for PKC in synaptic plasticity in this system. In particular, we have shown that distinct isoforms mediate distinct types of synaptic plasticity induced by the same neurotransmitter: The novel calcium-independent PKC Apl II is required for actions mediated by serotonin (5 HT) alone, while the classical calcium-dependent PKC Apl I is required for actions mediated when 5-HT is coupled to activity. We will discuss the reasons for PKC isoform specificity, assess the tools used to uncover isoform specificity, and discuss the implications of isoform specificity for understanding the roles of PKC in regulating synaptic plasticity. PMID- 17404387 TI - Pooled analysis and meta-analysis of the glutathione S-transferase P1 Ile 105Val polymorphism and bladder cancer: a HuGE-GSEC review. AB - The glutathione S-transferase P1 genotype (GSTP1) is involved in the inactivation of cigarette smoke carcinogens, and sequence variation in the gene may alter bladder cancer susceptibility. To examine the association between GSTP1Ile 105Val and bladder cancer, the authors undertook a meta- and pooled analysis. Summary crude and adjusted odds ratios and corresponding 95% confidence intervals were pooled by using a random-effects model. In the meta-analysis (16 studies, 4,273 cases and 5,081 controls), the unadjusted summary odds ratios for GSTP1 Ile/Val and Val/Val compared with GSTP1 Ile/Ile were 1.54 (95% confidence interval: 1.21, 1.99; p < 0.001) and 2.17 (95% confidence interval: 1.27, 3.71; p = 0.005). The association appeared to be the strongest in Asian countries. When the analysis was limited to European descendents (nine studies), the summary odds ratio decreased (odds ratio = 1.24, 95% confidence interval: 1.00, 1.52) (Q = 17.50; p = 0.02). All relevant data previously contributed to the International Study on Genetic Susceptibility to Environmental Carcinogens were pooled (eight studies, 1,305 cases and 1,558 controls). The summary odds ratios were similar to the ones from the meta-analysis. Case-only analyses did not detect an interaction between the GSTP1 genotype and smoking status (never/ever). GSTP1 Ile 105Val appears to be associated with a modest increase in the risk of bladder cancer. PMID- 17404388 TI - Anti-Toxoplasma gondii antibodies in patients with schizophrenia--preliminary findings in a Turkish sample. AB - Schizophrenia is a serious neuropsychiatric disease of uncertain etiology. We investigated the seropositivity rate for anti-Toxoplasma IgG and IgM antibodies by enzyme-linked immunosorbent assay (ELISA) in patients with schizophrenia to ascertain a possible relationship between Toxoplasma gondii and schizophrenia. We selected 100 patients with schizophrenia, 50 with depressive disorder, and 50 healthy volunteers to investigate the seropositivity rate of anti-Toxoplasma antibodies by ELISA. The seropositivity rate for anti-Toxoplasma IgG antibodies among schizophrenia patients (66%) was significantly higher than among patients with depressive disorder or healthy volunteers (P < .01). Thus, there might be a causal relationship between toxoplasmosis and the etiology of schizophrenia. PMID- 17404389 TI - Declining transition rate in ultra high risk (prodromal) services: dilution or reduction of risk? AB - There is growing interest in the prodromal stage of psychotic disorders, with many services now providing care for these "ultra high risk" (UHR) individuals. However, a reduction in the rate of transition to psychosis has been suspected over the last few years. This has implications for the use of interventions in this population and for the validity of research findings. This study examined the transition rate in one UHR service, the Personal Assessment and Crisis Evaluation Clinic, over the years 1995-2000 and investigated possible causes for the transition rate reduction. There was evidence for a declining transition rate, with each successive year showing a rate 0.80 times that of the preceding year. Functioning and symptom level were not responsible for the change. The decreased transition rate was partly explained by a reduction in the duration of symptoms of patients prior to receiving help. That is, UHR individuals are being detected and provided with care earlier than in the past. Thus, the decline in transition rate may be due to treatment being more effective at this very early stage of illness or it may be due to finding more false positives who were never at risk of psychosis, ie, a "dilution" effect. Given that it is not possible to distinguish between these alternatives at least phenotypically at present, perhaps it is time to rethink the role and practice of UHR clinics. Patients presenting to them need help. It may be that we need to aim to prevent a range of target syndromes. PMID- 17404390 TI - Wavelength-dependent modulation of brain responses to a working memory task by daytime light exposure. AB - In addition to classical visual effects, light elicits nonvisual brain responses, which profoundly influence physiology and behavior. These effects are mediated in part by melanopsin-expressing light-sensitive ganglion cells that, in contrast to the classical photopic system that is maximally sensitive to green light (550 nm), is very sensitive to blue light (470-480 nm). At present, there is no evidence that blue light exposure is effective in modulating nonvisual brain activity related to complex cognitive tasks. Using functional magnetic resonance imaging, we show that, while participants perform an auditory working memory task, a short (18 min) daytime exposure to blue (470 nm) or green (550 nm) monochromatic light (3 x 10(13) photons/cm2/s) differentially modulates regional brain responses. Blue light typically enhanced brain responses or at least prevented the decline otherwise observed following green light exposure in frontal and parietal cortices implicated in working memory, and in the thalamus involved in the modulation of cognition by arousal. Our results imply that monochromatic light can affect cognitive functions almost instantaneously and suggest that these effects are mediated by a melanopsin-based photoreceptor system. PMID- 17404391 TI - A pre-joining fitness test improves pass rates of Royal Navy recruits. AB - BACKGROUND: Military training represents a significant physical challenge. Low fitness levels are strongly associated with an increased risk of injury and training attrition. To increase pass rates for Royal Navy (RN) Phase I training, a pre-joining fitness test (PJFT) was introduced (2.4 km treadmill best effort run). The PJFT was designed to identify candidates with poor levels of aerobic fitness who may be prone to leave training prematurely. AIM: To examine the impact of the PJFT on training length and outcomes. METHODS: Time taken to run 2.4 km and training outcomes were measured before and after the introduction of the PJFT. Information was collected from RN Phase I training establishments and the network of careers offices between 2002 and 2005. Recruits were placed into quartile groups based on 2.4 km overground running performance. The no PJFT and PJFT groups and the 2.4-km run performance quartiles were compared for training outcomes and time spent in training. RESULTS: Training measures were available for 4818 recruits who entered training before the PJFT's introduction (no PJFT) and 3305 after its introduction (PJFT). The pass rate increased from 78 to 88% following the introduction of the PJFT (P < 0.01). The number of recruits applying for voluntary release decreased from 15% (no PJFT group) to 6% (PJFT group) (P < 0.05). CONCLUSION: The PJFT positively impacted on RN Phase 1 training pass rates. A greater number of recruits successfully completed training, fewer applied for voluntary release and the number and length of training extensions were reduced. PMID- 17404392 TI - Reduction of oxidative stress by compression stockings in standing workers. AB - BACKGROUND: Healthy workers who stand for prolonged periods show enhanced production of reactive oxygen species (ROS) in their systemic circulation. Oxidative stress is thought to be a risk factor for chronic venous insufficiency and other systemic diseases. AIM: To evaluate the effectiveness of compression stockings in the prevention of oxidative stress at work. METHODS: ROS and venous pressure of the lower limbs were measured in 55 theatre nurses who stood in the operating theatre for >6 h, 23 industrial ironers who stood for up to 5 h during their shift and 65 outpatient department nurses and 35 laundry workers who acted as controls. Subjects and controls were examined on two consecutive days before and after work and with and without compression stockings. RESULTS: Without compression stockings, lower limb venous pressure increased significantly after work in all subjects and controls (P < 0.001), while only operating theatre nurses showed significantly higher mean levels of ROS (P < 0.001). There was no significant difference in venous pressures and ROS levels after work in subjects or controls when wearing compression stockings. CONCLUSIONS: Our data suggest a preventive role of compression stockings against oxidative stress in healthy workers with a standing occupation. PMID- 17404393 TI - Band saw injury in a butcher. AB - BACKGROUND: While treating an unusual amputation caused by a meat band saw in a 35-year-old butcher, we sought information from the medical literature that would be useful to other physicians who might encounter similar occupational injuries. METHODS: Using the Medline database and relevant search terms, we reviewed the literature concerning occupational saw blade injuries and porcine microbiology as they related to this injury. RESULTS: Among meat workers using powered cutting equipment, hand injuries and distal fingertip amputations appear to be common. The greatest risk for a wound infection after open exposure to raw pork meat appears primarily related to environmental flora rather than enteric-borne porcine pathogens. CONCLUSIONS: Decision-making strategy when formulating a treatment plan for debridement or reconstruction of saw blade amputations should rely on a detailed understanding of the injury and occupational environment to achieve an optimal patient outcome. When considering operative and antibiotic treatment for porcine meat-related amputation injury, surgeons should adhere to open fracture-related guidelines, since porcine-borne illnesses are most often caused by ingestion rather than transcutaneous inoculation. PMID- 17404394 TI - Migration of intradermally injected quantum dots to sentinel organs in mice. AB - Topical exposure to nanoscale materials is likely from a variety of sources including sunscreens and cosmetics. Because the in vivo disposition of nanoscale materials is not well understood, we have evaluated the distribution of quantum dots (QDs) following intradermal injection into female SKH-1 hairless mice as a model system for determining tissue localization following intradermal infiltration. The QD (CdSe core, CdS capped, poly[ethylene glycol] coated, 37 nm diameter, 621 nm fluorescence emission) were injected intradermally (ID) on the right dorsal flank. Within minutes following intradermal injection, the highly UV fluorescent QD could be observed moving from the injection sites apparently through the lymphatic duct system to regional lymph nodes. Residual fluorescent QD remained at the site of injection until necropsy at 24 h. Quantification of cadmium and selenium levels after 0, 4, 8, 12, or 24 h in multiple tissues, using inductively coupled plasma mass spectrometry (ICP-MS), showed a time-dependent loss of cadmium from the injection site, and accumulation in the liver, regional draining lymph nodes, kidney, spleen, and hepatic lymph node. Fluorescence microscopy corroborated the ICP-MS results regarding the tissue distribution of QD. The results indicated that (1) ID injected nanoscale QD remained as a deposit in skin and penetrated the surrounding viable subcutis, (2) QD were distributed to draining lymph nodes through the sc lymphatics and to the liver and other organs, and (3) sentinel organs are effective locations for monitoring transdermal penetration of nanoscale materials into animals. PMID- 17404395 TI - Induction of a unique gene expression profile in primary human hepatocytes by hepatitis C virus core, NS3 and NS5A proteins. AB - Hepatocellular carcinoma (HCC) is a fatal disease and hepatitis B and C viruses (HBV and HCV) are considered as major causative factors for the development of HCC. We have conducted gene expression profiling studies to search for potential target genes responsible for HCV-mediated HCC. Adenoviruses encoding core (HCV structural protein), NS3 and NS5A [HCV non-structural (NS) proteins] were generated and infected individually or together in freshly isolated primary human hepatocytes. An adenovirus harboring the oncogenic HBV protein, HBx, was included for comparison. A microarray platform of over 22,000 human oligos was analyzed to seek out significant differentially expressed genes among these viral proteins. We also compared these gene expression profiles with those obtained from HCV infected liver samples from chronic liver disease (CLD) patients and HCV-related HCC. We found that HCV-related proteins largely induce unique genes when compared with HBx. In particular, interferon-inducible gene 27 (IFI27) was highly expressed in HCV or core-infected hepatocytes and HCV-related CLD or HCC, but was not significantly expressed in HBx-infected hepatocytes or HBV-related CLD or HCC, indicating that IFI27 may play a role in HCV-mediated HCC. In conclusion, our results suggest that HBV and HCV promote HCC development mainly through different mechanisms. PMID- 17404396 TI - Involvement of ERKs, RSK2 and PKR in UVA-induced signal transduction toward phosphorylation of eIF2alpha (Ser(51)). AB - Double-stranded RNA-dependent protein kinase R (PKR) has been implicated in anti viral (antitumor) and apoptotic responses. PKR is activated by extracellular stresses and phosphorylates the alpha subunit of protein synthesis initiation factor eIF2, thereby inhibiting protein synthesis and impeding virus multiplication. Phosphorylation of eIF2alpha in mammalian cells has been shown to be increased after ultraviolet (UV) stress and to be required for UV-induced repression of protein translation. UVA is an important etiological factor in skin carcinogenesis and we observed that UVA induced phosphorylation of PKR (Thr(451)) and eIF2alpha (Ser(51)) in mouse skin epidermal JB6 Cl41 cells. The induction was suppressed by the MEK1 inhibitor, PD 98059. UVA stimulation of PKR and eIF2alpha phosphorylation was also inhibited by a dominant-negative mutant (DNM) of ERK2- or RSK2-deficient cells (RSK2(-)). An inhibitor of p38, SB 202190 or a DNM of p38alpha kinase (DNM-p38alpha) suppressed UVA-induced phosphorylation of eIF2alpha (Ser(51)) but had no effect on phosphorylation of PKR (Thr(451)). Our data indicated that phosphorylation of PKR at Thr(451) is mediated through ERK2 and RSK2, but not through p38 kinase, and is involved in the regulation of Ser(51) phosphorylation of eIF2alpha in UVA-irradiated JB6 cells. In vitro and in vivo kinase assays indicated that phosphorylation of eIF2alpha at Ser(51) occurred indirectly through ERK2, RSK2 or p38 kinase in the cellular response to UVA. These data may lead to the use of these signaling molecules as targets to develop more effective chemopreventive agents with fewer side effects to control UV-induced skin cancer. PMID- 17404397 TI - Analysis of nuclear copies of mitochondrial sequences in honeybee (Apis mellifera) genome. AB - At least 0.08% of the Apis mellifera nuclear genome contains sequences that originated from mitochondria. These nuclear copies of mitochondrial sequences (numts) are scattered all over the honeybee chromosomes and have originated by multiple independent insertions of mitochondrial DNA (mtDNA) as evident by phylogenetic analysis. Apart from original insertions, moderate duplications of numts also contributed to the present pattern and distribution of mitochondrial sequences in honeybee chromosomes. Assimilation of mitochondrial genes in the nuclear genome is mediated by extensive fragmentations of the original inserts. Replication slippage seems to be a major mechanism by which small sequences are inserted or deleted from mtDNA destined to nucleus. Most of the honeybee numts (84%) are located in the nongenic regions. The majority (94%) of the numts that are located in predicted nuclear genes have originated from mitochondrial genes coding for cytochrome oxidase and NADH dehydrogenase subunits. On the other hand, the mitochondrial rRNA or tRNA gene sequences are predominantly (88%) located in nongenic regions of the genome. Evidences also support for exertion of purifying selection on numts located in specific genes. Comparative analysis of numts of European, African, and Africanized honeybees suggests that numt evolution in A. mellifera is probably not demarked by speciation time frame but may be a continuous and dynamic process. PMID- 17404398 TI - The genetic architecture of ecological speciation and the association with signatures of selection in natural lake whitefish (Coregonus sp. Salmonidae) species pairs. AB - Adaptive evolutionary change is contingent on variation and selection; thus, understanding adaptive divergence and ultimately speciation requires information on both the genetic basis of adaptive traits as well as an understanding of the role of divergent natural selection on those traits. The lake whitefish (Coregonus clupeaformis) consists of several sympatric "dwarf" (limnetic) and normal (benthic) species pairs that co-inhabit northern postglacial lakes. These young species pairs have evolved independently and display parallelism in life history, behavioral, and morphological divergence associated with the use of distinct trophic resources. We identified phenotype-environment associations and determined the genetic architecture and the role of selection modulating population genetic divergence in sympatric dwarf and normal lake whitefish. The genetic architecture of 9 adaptive traits was analyzed in 2 hybrid backcrosses individually phenotyped throughout their life history. Significant quantitative trait loci (QTL) were associated with swimming behavior (habitat selection and predator avoidance), growth rate, morphology (condition factor and gill rakers), and life history (onset of maturity and fecundity). Genome scans among 4 natural sympatric pairs, using loci segregating in the map, revealed a signature of selection for 24 loci. Loci exhibiting a signature of selection were associated with QTL relative to other regions of the genome more often than expected by chance alone. Two parallel QTL outliers for growth and condition factor exhibited segregation distortion in both mapping families, supporting the hypothesis that adaptive divergence contributing to parallel reductions of gene flow among natural populations may cause genetic incompatibilities. Overall, these findings offer evidence that the genetic architecture of ecological speciation is associated with signatures of selection in nature, providing strong support for the hypothesis that divergent natural selection is currently maintaining adaptive differentiation and promoting ecological speciation in lake whitefish species pairs. PMID- 17404400 TI - Drug-eluting stent implantation in coronary trifurcation lesions. AB - BACKGROUND: There is no specific study evaluating the outcome of DES implantation in trifurcation lesions. OBJECTIVE: To evaluate the mid-term clinical and angiographic outcome of drug-eluting stent (DES) implantation in trifurcation lesions. METHODS: All complications and major adverse cardiac events, including cardiac death, Q-wave myocardial infarction (MI), target lesion revascularization (TLR), and target vessel revascularization (TVR) were recorded in-hospital and during clinical follow up. RESULTS: A total of 15 consecutive patients undergoing percutaneous coronary intervention with DES in de novo trifurcation lesions were identified. Lesions were located as follows: 13 (86.7%) at the distal left main coronary artery (LMCA) comprising the left anterior descending artery (LAD), the left circumflex artery (LCX) and an intermediate branch; 1 between the LAD, diagonal, and septal branches; and 1 between the LCX, obtuse marginal and posterior lateral branches. Stenting was performed in all 3 branches in 8 patients, in 2 branches in 6 patients, and in 1 branch in 1 patient. The mean follow-up period was 19.0 +/- 8.3 months. TLR occurred in 3 patients (20%) with LMCA lesions. TVR occurred in 6 patients (40%). Of those, 3 were due to TLR, while the other 3 for progression of nontarget lesions. No deaths, Q-wave MIs or stent thromboses were recorded. CONCLUSION: Most trifurcation lesions were found in the distal LMCA. DES implantation in trifurcation lesions can be performed with a low incidence of death, Q-wave MI or stent thrombosis. PMID- 17404401 TI - Propensity score analysis of vascular complications after diagnostic cardiac catheterization and percutaneous coronary intervention using thrombin hemostatic patch-facilitated manual compression. AB - OBJECTIVES: To evaluate the adjusted risk of vascular complications after thrombin hemostasis patch-facilitated manual compression (THP-MC) for femoral artery access site management. BACKGROUND: Thrombin hemostatic patches shorten time to hemostasis after cardiac procedures involving femoral artery access, but whether these patches are as safe as manual compression remains uncertain. METHODS: THP-MC (D-Stat Dry, Vascular Solutions, Minneapolis, Minnesota) was used in 3,464 consecutive patients including 2,464 diagnostic cardiac catheterizations (CATH) and 1,000 percutaneous coronary intervention procedures (PCI) performed via a femoral access at a single site (WFUBMC). A total of 4,371 procedures including 2,956 CATH and 1,415 PCI performed prior to use of THP-MC, and treated with manual compression, served as the control group. Ambulation was permitted 2 to 3 hours after THP-MC, and 3 to 8 hours after MC. Propensity to receive a THP was calculated, and adjusted in-hospital outcomes evaluated. RESULTS: Time to hemostasis was similar for THP-MC (13.0 +/- 3.3 min) compared to MC (14.4 +/- 5.7 min), p = 0.51 for CATH, and was shorter for THP-MC (14.2 +/- 5.4 minutes) compared to MC (20.1 +/- 5.4 min), p <0.001, for PCI. Overall, the unadjusted incidence of any vascular complication (VC) was 0.6+/- for THP-MC and 1.0% for manual compression, p = 0.036. The propensity score-adjusted odds ratio for any VC comparing THP-MC to manual compression was 0.58 (0.34 to 0.99) for all procedures, 0.42 (0.20 to 0.87) for diagnostic procedures and 0.86 (0.39 to 1.88) for PCI. CONCLUSIONS: In this large, single-center, contemporary observational study, use of THP-MC shortened manual compression time for PCI procedures, permitted early ambulation and was as safe as conventional manual compression. PMID- 17404402 TI - Comparison of vessel response following sirolimus-eluting stent implantation as assessed by serial 3-D intravascular ultrasound study. AB - Recent sirolimus-eluting stent (SES) studies have suggested higher rates of restenosis in non-left anterior descending (LAD) artery lesions. The aim of this study was to evaluate differential vessel response (LAD versus non-LAD) to SES implantation using serial intravascular ultrasound (IVUS). A total of 94 patients who underwent SES implantation and serial (post-PCI and 8 months) 3-dimensional IVUS were enrolled from our database. Volumetric analysis was performed throughout the stent as well as the adjacent reference segment (up to 5 mm). Volume index (volume/length) was calculated for vessel (VVI), lumen (LVI), and plaque (PVI). Cross-sectional narrowing (CSN) was defined as neointimal area divided by stent area (%). With respect to the in-stent segment, VVI, PVI, and LVI at post-PCI were not significantly different between the LAD (n = 41) and non LAD (n = 53) lesions. At follow up, however, maximum CSN was significantly greater in the non-LAD lesions (18.3 +/- 15.2% versus 12.2 +/- 10.0%; p = 0.029). At the proximal reference segment, the non-LAD lesions showed a significantly greater LVI decrease than the LAD lesions (p <0.05), primarily due to mild vessel shrinkage observed in the non-LAD lesions. There were no significant differences at the distal reference segment between the LAD and non-LAD lesions. This detailed IVUS analysis suggests that there are minimal differences in the vessel responses following SES implantation. These findings may have potential implications for mechanical and pharmacokinetic properties of next-generation drug-eluting stent technology. PMID- 17404403 TI - Sirolimus-eluting stents for the treatment of bare-metal in-stent restenosis: long-term clinical follow up. AB - BACKGROUND: Studies focusing on short- and mid-term follow up support the beneficial role of sirolimus-eluting stents (SES) in the treatment of in-stent restenosis (ISR), yet no long-term safety and/or efficacy data are available. METHODS: Patients with ISR following bare-metal stenting (BMS) and treated with SES were prospectively studied. Baseline, procedural, and in-hospital data were appraised. The primary endpoint was the rate of major cardiovascular events (MACE) at long-term follow up (>9 months). Secondary endpoints were the individual contributors to MACE. RESULTS: A total of 180 SES were implanted to treat 138 consecutive patients. Procedural success was achieved in all patients without in-hospital death, acute stent thrombosis, stroke, or urgent coronary artery bypass. During follow up, MACE occurred in 5.8% of patients at 6 months, 14.3% at 12 months, and 25% at 24 months. Specifically, all-cause mortality was 1.7% at 6 months, 3.5% at 12 months, and 4.8% at 24 months, for a total of 5 deaths. Target vessel revascularization occurred at 6, 12, and 24 months in 4.2%, 11.2%, and 15.9% of patients, respectively, while target lesion revascularization (TLR) alone accounted for 3.4% at 6 months, 9.6% at 12 months, and 11% at 24 months. Three case of myocardial infarction occurred during follow up (2.2%), without any surgical revascularization or stent thrombosis. CONCLUSIONS: Treatment of ISR with SES appears safe and effective, even if a 10% annual rate of MACE can be expected, with a sizable portion of these due to apparently nontarget lesion events. PMID- 17404404 TI - Sirolimus-eluting, bioabsorbable polymer-coated constant stent (Cura) in acute ST elevation myocardial infarction: a clinical and angiographic study (CURAMI Registry). AB - BACKGROUND: There are safety concerns over the current polymer-based drug-eluting stents (DES) on the possible delayed healing process and adverse reactions to the polymer when drug elution is completed. Cura (Orbus Neich) is a sirolimus eluting, bioabsorbable, polymer-coated constant stent. METHODS: From March 1 to June 30, 2005, Cura stent implantation was instituted as the default strategy for percutaneous coronary intervention in 49 consecutive ST-Elevation Myocardial Infarction (STEMI) patients (male 86%; average age 55 +/- 10 years; diabetes 31%). RESULTS: The angiographic success rate was 100%. In-hospital adverse events consisted of 1 in-hospital death (2% heart failure). A total of 27 patients (56%) underwent 8-month angiographic follow up. Binary restenosis occurred in 6 patients (22%), and late loss was 0.74 +/- 0.89 mm. At 9-month follow up, a total of 5 patients had 6 major adverse events (1 death, 1 reinfarction, and 4 target lesion revascularizations). There was no incidence of stent thrombosis. CONCLUSION: Cura stent implantation appears to be feasible and safe in the treatment of STEMI. However, the binary restenosis rate and late loss at angiographic follow up were similar to that of bare-metal stents, and therefore compare unfavorably with other proven benchmark DES. PMID- 17404405 TI - Is the bioabsorbable stent an answer for late stent thrombosis. PMID- 17404406 TI - DK crush (double-kissing and double-crush) technique for treatment of true coronary bifurcation lesions: illustration and comparison with classic crush. AB - BACKGROUND: Classic crush has a lower success rate compared to final kissing balloon inflation (FKBI). We previously reported the double-kissing (DK) crush technique that involves double-kissing along with double-crushing for the treatment of true bifurcation coronary lesions in 2005. METHODS AND RESULTS: This is a consecutive, nonrandomized, open-label study. Eighty-eight consecutive patients with single, true coronary bifurcation lesions according to Lefevre Classification2 and side branch diameter >2.0 mm were enrolled. The first 44 patients (from October 2004 to January 2005) were assigned to the classic crush treatment arm and the next 44 patients (from February 2005 to June 2005) were assigned to the DK crush technique arm, respectively. Data within 30 days were analyzed. Patients in the DK crush group, compared to those in classic crush group, were characterized by longer lesion length in the side branch (13.5 +/- 3.4 mm vs 7.8 +/- 3.1 mm; p <0.05), shorter procedural time (44 +/- 12 minutes vs 68 +/- 17 minutes; p <0.05 ), higher success rate of FKBI (100% vs 70%; p <0.01), and lower contrast volume required (102 +/- 38 mm vs 176 +/- 46 mm; p <0.05). There was a trend toward a larger distal angle between the main vessel and the side branch (57 +/- 180 vs 47 +/- 150) in the DK crush group, as well as longer lesion length in the main vessel (24.3 +/- 8.6 mm vs 21.1 +/- 7.3 mm), though without significant differences (p >0.05). Subacute stent thrombosis was detected in 2 patients with failure of FKBI in the classic crush group (4.3%). In addition, patients in the classic crush group were characterized by a smaller minimum lumen diameter (MLD) at the side branch ostium (2.74 +/- 0.12 mm vs 3.01 +/- 0.13 mm; p <0.01) and had a higher degree of residual stenosis at the ostial side branch (17.4 +/- 11.2% vs 7.3 +/- 8.6%; p <0.05). CONCLUSIONS: Compared with classic crush, DK crush has the potential to improve the clinical outcome in patients with coronary bifurcation lesions. Further randomized, prospective, multicenter studies are required to confirm these differences between the classic crush and DK crush techniques. PMID- 17404407 TI - Buried wire technique: enhancing support method for complex percutaneous interventions and stenting. PMID- 17404408 TI - Transseptal TandemHeart implantation through an Amplatzer atrial septal occluder. PMID- 17404409 TI - Kounis syndrome: a potential cause of simultaneous multivessel coronary spasm and thrombosis after drug-eluting stent implantation. PMID- 17404410 TI - Spontaneous multivessel coronary vasospasm leading to cardiogenic shock. AB - We report a case of spontaneous multivessel coronary vasospasm leading to anterior myocardial infarction and cardiogenic shock in the setting of treatment with methylphenidate and withdrawal from beta-blockers and calcium channel antagonists. The patient was stabilized with an intra-aortic balloon pump and treated with coronary stenting and vasodilators and ultimately had an uneventful recovery. We review the diagnosis and treatment of coronary vasospasm and the implications of coronary vasoreactivity in the pathophysiology of myocardial infarction. PMID- 17404411 TI - Methamphetamine-associated acute myocardial infarction and cardiogenic shock with normal coronary arteries: refractory global coronary microvascular spasm. AB - Methamphetamine (MET) is a growing public health concern and is prevalent in, although not limited to, the youth. The drug's association with myocardial infarction is well described and is attributed to accelerated atherosclerosis, hypercoagulable state, and macrovascular epicardial coronary spasm. However, global slow-flow of all coronary systems in the absence of significant stenoses has not been previously reported. We hereby present a young patient who likely experienced severe, global microvascular coronary spasm unrelieved by intracoronary vasodilator therapy, resulting in acute myocardial infarction. The pharmacology of MET, its postulated mechanism in acute coronary syndromes, as well as the pathophysiology and treatments of microvascular coronary spasm are briefly reviewed. Readers are recommended to be vigilant of potential illicit drug use in patients with atypical presentations of acute coronary syndromes. PMID- 17404412 TI - Catheter-induced left main coronary artery dissection resulting in abrupt closure and cardiac arrest: successful stenting during resuscitation. AB - Catheter-induced left main coronary artery dissection is an uncommon but devastating complication of coronary angiography and percutaneous coronary intervention. We present a case of left main coronary artery dissection induced with a guide catheter, which resulted in acute occlusion and cardiac arrest. Survival and complete functional recovery were achieved with bailout stenting. PMID- 17404413 TI - Single coronary artery from right aortic sinus with septal course of left anterior descending artery and left circumflex artery as continuation of right coronary artery: a hitherto unreported coronary anomaly. AB - A single coronary artery (SCA) in the absence of structural heart disease is a rare coronary anomaly and is often detected incidentally during coronary angiography. We report a hitherto undescribed type of SCA originating from the right sinus of Valsalva, with the left anterior descending artery having a septal course and the right coronary artery continuing as the left circumflex artery, which was incidentally detected in a 73-year-old female. PMID- 17404414 TI - A novel method of clot extraction using a FilterWire EX in acute myocardial infarction. AB - We describe a novel approach for the use of a FilterWire EX distal protection device as a snaring device for clot extraction in a patient who sustained acute ST-elevation myocardial infarction due to occlusion of the right coronary artery. PMID- 17404415 TI - Extreme coronary guide catheter support: a case of a novel telescopic guide catheter system with a contralateral aortic wall support. AB - Extra support of the guide catheter is necessary in some cases of percutaneous coronary intervention (PCI). We describe a successful case of PCI of a very calcified and tortuous right coronary artery in which a modification of a novel telescopic guide system was applied. A long sheath that armored the guide catheter allowed extreme support derived from the contralateral aortic wall. The operator can adjust the support of the guide system from soft to extremely stiff. PMID- 17404416 TI - Late stent thrombosis associated with coronary aneurysm formation after sirolimus eluting stent implantation. AB - Stent thrombosis is a rare but potentially fatal complication of coronary stent implantation. Its occurrence late after drug-eluting stent (DES) deployment has led to concerns regarding their long-term safety. We report a case of late stent thrombosis 26 months after sirolimus-eluting stent (SES) (Cypher, Cordis Corp., Miami, Florida) implantation. This was associated with marked positive vessel remodeling and coronary aneurysm formation involving the stented segment of the coronary artery. The patient was on dual antiplatelet therapy at the time. PMID- 17404417 TI - Complete Cypher stent fracture and migration in the ostium of the right coronary artery. PMID- 17404418 TI - Characterization of activation energy distribution for ultraviolet irradiation of T-rich oligonucleotide using high-performance liquid chromatography electrospray ionization mass spectrometry tandem mass spectrometry. AB - Exposure to solar UV radiation gives rise to mutations that may lead to skin cancer of human being. Series of experiments were carried out in order to reveal activation energy distribution of DNA mutation caused by UV radiation. The T-rich oligonucleotides were exposed to UV radiation with increasing intensity for different durations. Photoproducts of T-rich oligonucleotide were investigated using ion-pair reversed-phase high-performance liquid chromatography/tandem electrospray ionization mass spectrometry (IP-RP-HPLC/ESI-MS) at room temperature. Two photoproducts of T-rich oligonucleotide were cis-syn cyclobutane pyrimidine dimmer (T[c,s]T) and the pyrimidine(6,4)pyrimidone product (T[6,4]T). Activation energy distribution of DNA mutation was calculated using a commercial kinetics analysis programs by Robert L. Braun and Alan K. Burnham , Lawrance Livermore International Laboratory (version 2.4.1). To use the software for deriving the kinetics parameters, the factor T (temperature) in the software was substituted with k1R, in which k1 is a factor, R is radiation intensity. The activation energy derived ranges from 55 to 110 kJ mol(-1). By the same software, those kinetics parameters were extrapolated to natural UV radiation process to predict DNA damage degree without the DNA repair process. PMID- 17404419 TI - Determination of dissociation constants of cyclodextrin-ligand inclusion complexes by electrospray ionization mass spectrometry. AB - The inclusion complexes of four ligands binding to cyclodextrins (CDs) were studied by electrospray ionization mass spectrometry (ESI-MS) and the dissociation constants of the complexes were obtained. The 1:1 stoichiometric inclusion complex was found in the system of CD and fenbufen or aspirin. The obtained KD values of the inclusion complexes of fenbufen binding to alpha-CD and to beta-CD are 4.38x10(-4) mol L(-1) and 2.12x10(-4) mol L(-1), respectively. The KD values of the inclusion complexes of alpha-CD-aspirin and beta-CD-aspirin are 3.33x10(-4) mol L(-1) and 1.83x10(-4) mol L(-1), respectively. A non-linear least squares regression method was applied to validate the results which were consistent with each other. For the system of tetracycline hydrochloride and CD, the 1:1 and 1:2 stoichiometric inclusion complexes were found in the mass spectra. The KD,1 and KD,2 values of the 1:1 and 1:2 stoichiometric inclusion complexes of alpha-CD and tetracycline hydrochloride are 4.47x10(-4) mol L(-1) and 6.51x10(-4) mol L(-1), respectively, and those of beta-CD and tetracycline hydrochloride are 2.26x10(-4) mol L(-1) and 8.57x10(-4) mol L(-1), respectively. For the system of norfloxacin and CD, besides the 1:1 and 1:2 inclusion complexes, the 1:3 stoichiometric inclusion complex was also found. The KD,1, KD,2 and KD,3 of alpha-CD and norfloxacin inclusion complexes are 4.61x10(-4) mol L(-1), 6.05x10(-4) mol L(-1) and 1.45x10(-3) mol L(-1), respectively. The three KD values of beta-CD and norfloxacin are 1.96x10(-4) mol L(-1), 4.93x10(-4) mol L(-1) and 1.15x10(-3) mol L(-1), respectively. PMID- 17404420 TI - A comparison of electrospray-ionization and matrix-assisted laser desorption/ionization mass spectrometry with nuclear magnetic resonance spectroscopy for the characterization of synthetic copolymers. AB - Electrospray ionization (ESI-MS) and matrix assisted laser desorption-ionization (MALDI-MS) were used to determine the composition (monomer ratios) and structure (end group analysis) relative to 1H NMR spectroscopy and theoretical predictions for three different copolymers: poly(butyl acrylate/vinyl acetate) (PBA/PVAc), poly(methyl methacrylate/vinyl acetate) (PMMA/PVAc) and poly(butyl acrylate/methyl methacrylate) (PBA/PMMA). We found that the ESI results were in excellent agreement with 1H NMR spectroscopy for PBA/PVAc and PBA/PMMA copolymers whereas there was more divergence in the case of PMMA/PVAc. In the case of PBA/PMMA copolymers similar distributions of products were observe by ESI-MS and MALDI-MS with the two major products classes differing by their end-groups. One class has hydrogen and dodecylthio end groups while in the other the dodecylthio has been replaced by alpha-cyanoisopropyl from the initiator. The relative abundance of these distributions as a function of copolymer conversion for a series of reaction conditions was investigated by both ESI and MALDI. MALDI results consistently underestimated (relative to ESI) the butylacrylate monomer ratio in PBA/PMMA and the abundance of co-polymer oligomers terminated by a dodecylthio group from the chain transfer agent. PMID- 17404421 TI - Influence of solvent and counter ion on copper complexes with N-alkyl-pyridine-2 carboxamides as studied by electrospray ionization mass spectrometry. AB - The interactions of copper(II) with N-alkyl-pyridine-2-carboxamides (M) were studied by electrospray ionization mass spectrometry. The influence of solvent and counter ion (Cl-, ClO4-) on the type of ions (complexes) observed was discussed. By cone voltage increase, the fragmentation "in source" of the ions discussed was achieved. In methanol solution containing N-alkyl-pyridine-2 carboxamide (M) and CuCl2 the singly- and doubly charged-ions [M+CuCl]+ and [M2+Cu]+2 were detected. In acetonitrile solution containing N-alkyl-pyridine-2 carboxamide and CuCl2, the copper(I)-containing ions [M+CH3CN+Cu]+ were formed. The use of Cu(ClO4)2 instead of CuCl2 resulted in more abundant doubly-charged ions in both methanol and acetonitrlile solutions and, for the former solution, the ions containing methoxyl anion, namely [M+CuOCH3]+, were observed. When water was used as a solvent, the abundant ions corresponding to the protonated ligands were formed, while the ions corresponding to copper complexes were characterised by low abundances. PMID- 17404422 TI - Mass spectrometry of beta-ketoesters. Some evidence of their tautomerism. AB - Mass spectrometric evidence of tautomerism is reported for beta-ketoesters. The analysis of the corresponding mass spectra has allowed specific assignment of fragment ions to tautomers. The predictive value of this methodology is supported by the influence of substitution pattern of these compounds on these equilibria. Experimental data are strongly supported by Austin Model 1 semiempirical calculations indicating that mass spectrometry could be resourced as a tool for the investigation of tautomerism of neutral species in the gas phase. PMID- 17404423 TI - In vitro nitrosation of insulin A- and B-chains. AB - The physiological roles of insulin and nitric oxide (NO) have been recently recognized by several studies. A diversity of chemical modifications of insulin is reported both in vivo and in vitro. S-nitrosation, the covalent linkage of NO to cysteine free thiol is recognized as an important post-translational regulation in many proteins. Here we report the in vitro synthesis of an S nitrosothiol of bovine insulin A- and B-chains. These compounds were characterized by their HPLC chromatographic behavior, monitored by UV visible spectroscopy and electron spray ionization mass spectrometry. The experimental results indicate that each A- and B-chain were S- nitrosated with only one NO group. Stability and solubility of these synthesized derivatives is described for physiological purposes. In this work, nitroso A- and B-chains of insulin were synthesized in vitro in order to better understand the possible interactions between insulin and NO that may be involved in the etiology of insulin resistance. PMID- 17404424 TI - Letter: electron ionization-induced fragmentation of new thiochalcone derivatives. AB - Electron ionization mass spectral fragmentation of new (E)-4-chalconylothioalkyl substituted derivatives of piperidinie, 4-methylpiperidine, morpholine and piperazine as well as (E)-4-bromoalkylthiochalcones being substrates of the former compounds have been studied. Fragmentation pathways have been proposed on the basis of the accurate mass and metastable transition measurements. PMID- 17404426 TI - Equilibrium conditions in laser-desorbed plumes: thermodynamic properties of alpha-cyano-4-hydroxycinnamic acid and protonation of amino acids. AB - The equilibrium nature of a plume of laser desorbed material is explored through the application of a simple equilibrium model to the ion signals observed in 355 nm laser desorption/ionization mass spectra of mixtures of the MALDI matrix alpha cyano-4-hydroxycinnamic acid (alphaCHCA) with the amino acids glycine, alanine, valine, isoleucine, and phenylalanine. In these studies it is found that there are systematic and predictable increases in the relative yield of protonated amino acid with increases in amino acid gas-phase basicity. In addition, the thermodynamic values extracted from the equilibrium plot are shown to be in good agreement with values obtained from computational investigation of plausible alphaCHCA proton donor species. These results are supportive of a picture wherein the laser-desorbed material is viewed as a dense plume in which facile charge transfer occurs leading, ultimately, to a thermodynamically equilibrated distribution of proton donor and proton acceptor species. PMID- 17404427 TI - Gas-phase ionization/desolvation processes and their effect on protein charge state distribution under matrix-assisted laser desorption/ionization conditions. AB - The charge state distribution of proteins was studied as a function of experimental conditions, to improve the understanding of the matrix-assisted laser desorption/ionization (MALDI) mechanisms. The relative abundances of the multiply-charged ions appear to be a function of the matrix chosen, the laser fluence and the matrix-to-analyte molar ratio. A correlation is found between the matrix proton affinity and the yield of singly- versus multiply-charged ions. These results are in good agreement with a model in which gas-phase intracluster reactions play a significant role in analyte ion formation. A new model for endothermic desolvation processes in ultraviolet/MALDI is presented and discussed. It is based upon the existence of highly-charged precursor clusters and, complementary to the ion survivor model of Karas et al., assumes that two energy-dependent processes exist: (i) a soft desolvation involving consecutive losses of neutral matrix molecules, leading to a multiply-charged analyte and (ii) hard desolvation leading to a low charge state analyte, by consecutive losses of charged matrix molecules. These desolvations pathways are discussed in terms of kinetically limited processes. The efficiency of the two competitive desolvation processes seems related to the internal energy carried away by clusters during ablation. PMID- 17404429 TI - Application and evaluation of solvent-free matrix-assisted laser desorption/ionization mass spectrometry for the analysis of derivatized fullerenes. AB - A variety of derivatized fullerenes have been studied by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Of particular emphasis has been the evaluation of a recently introduced solvent-free sample/target preparation method. Solvent-free MALDI is particularly valuable in overcoming adverse solvent related effects, such as insolubility and/or degradation of the sample. The method was applied to fullerene derivatives susceptible to decomposition under insufficiently "soft" MALDI conditions. Analytes included the hydrofullerene: C(60)H(36), fluorofullerenes: C(60)F(x) where x = 18, 36, 46, 48 and C(70)F(x) where x = 54, 56, methano-bridged amphiphilic ligand adducts to C(60) and the [4 + 2] cycloadduct of tetracene to C(60). The new solvent-free sample preparation is established as an exceedingly valuable addition to the repertoire of preparation protocols within MALDI. The MALDI mass spectra were of very high quality throughout, providing a testimony that "soft" MALDI conditions could be achieved. Using the [4 + 2] cycloadduct of tetracene to C(60) as the model analyte for direct comparison with solvent-based MALDI, the solvent-free approach led to less fragmentation and more abundant analyte ions. Applying solvent-free sample preparation, different matrix compounds have been examined for use in the MALDI of derivatized fullerenes, including sulfur, tetracyanoquinodimethane (TCNQ), 9-nitroanthracene (9-NA) and trans-2-[3-(4-tert-butylphenyl)-2-methyl-2- propenylidene]malononitrile (DCTB). DCTB was confirmed as the best performing matrix, reducing unwanted decomposition and suppression effects. PMID- 17404430 TI - Nerve injuries in urological surgery. AB - It is well known that surgical procedures are followed by different complications, connected with nerve injury at the site of wound. In case of complete lesion due to nerve suturing or nerve interposition the prognosis remains questionable and may be complicated by neuroma later on. The symptoms of nerve damage are Causalgia (in case of combined with lesion of Nn. splanchnici: extreme burning pain, maximum after weeks, decreasing later on); Posttraumatic neuralgia: located mostly in legs: long persistent dump pains; and Neuroma Following complete lesion with or without nerve suture or nerve interposition: extremely painful and usually revision is necessary. During the surgical access to the kidneys n. Intercostalis 11-T12, n. Iliohypogastricus T12-L1, n. Ilioinguinalis T12-L1, n. Iliohypogastricus T12-L1, n. Ilioinguinalis T12-L1 may get damaged, in case of groin incision-n. Cutaneus femoralis lateralis L2-L3, n. Pudendalis S2-S4 and in Psoas Hitch procedure-n. Genitofemoralis L1-L2, n. Femoralis L1-L4, Obturatorius L2-L5. PMID- 17404431 TI - Prognostic significance of p27(Kip 1) expression in renal cell carcinoma. AB - The importance of cyclin-dependent kinase inhibitor genes (CDKIG) in benign and malignant urological diseases is a subject of intense ongoing investigation. The goal of the current study was to analyze the expression of p27(Kip 1) CDKIG in benign and malignant renal cells and assess their possible association with different clinical parameters. Expression of p27(Kip 1) CDKIG was evaluated and compared in 24 normal human kidneys and in 46 renal cell carcinoma (RCC) samples. Intensity of the gene expression was compared between the groups and possible association was analyzed with the cancer clinical parameters. The gene was significantly higher expressed in normal than in RCC tissue samples (p=0.0045). Intensity of the marker expression in RCC was negatively correlated with tumor size (Rho=-0.438, p=0.0051) and associated with stage and grade (p=0.0488 and <0.0001, respectively). The patients with symptomatic disease had significantly less marker expression than incidentally discovered tumors (p=0.0301). The marker expression was significantly higher in oncocytomas as compared with conventional RCCs (p=0.0378) The baseline p27(Kip 1) expression level in these patients was significantly lower than in non-recurrent tumors (p=0.04). Disease-related related death was observed in 4 cases. The baseline p27(Kip 1) expression level in these patients was significantly lower than in alive patients (p=0.0106). The Log-Rank analysis showed that loss of p27(Kip 1) expression were the risk-factors of oncological patient death (p=0.005). Expression of p27 is significantly decreased in RCC as compared with normal kidney tissue. Intensity of the gene expression is associated with clinical parameters: tumour size, stage, grade and disease presentation. Loss of p27 expression is a risk-factor for disease progression. PMID- 17404432 TI - Kidney transplantation in Georgia: a single centre experience with living-related donors. AB - 37 patients underwent living donor kidney Tx at the National Centre of Urology from January 2000 to December 2007. All transplants were performed from genetically related donors. The mean follow-up was 29,5 months. The following parameters were analysed: early and late posttransplant complications, rejection rate, the most recent Cr, patient and graft survival rates. Haemorrhage (5.4%), gastro-intestinal bleeding (10.8%) and graft thrombosis (5.4%) were the most important early complications. 6 (16.2%) episodes of acute cellular rejection were detected of whom 5 were treated successfully with bolus steroids. Two (5.4%) patients developed delayed graft function and needed postoperative dialyses. The mean value of the most recent serum Cr level is 1.6+/-1.1 g/dl (range: 0.7-6.2 g/dl). 1, 2 and 3 year patient survival for the whole group was 89.2%. 1, 2 and 3 year graft survival for the whole group was 89.2%, 83% and 83%, respectively. The mortality rate associated with kidney Tx has decreased from 38% in 1995-2000, to 5.4% in 2000-2007. The results of kidney Tx can achieve the acceptable level with improving surgical technique and experience. The pool of living-unrelated (spouses etc.) donors should be wider used to increase the number of transplants. PMID- 17404433 TI - Is radical prostatectomy an option in high-risk prostate cancer patients? AB - Long term survival of men with high-risk (T3 and/or PSA>20 ng/ml and/or Gleason>or=8) prostate cancer is still far from satisfaction. Common treatment options for these patients usually include hormonal therapy, radiation therapy or a combination of both. Radical prostatectomy has not been considered as the standard treatment strategy in these cases, while the potential for incomplete excision of the local tumor and high incidence of lymph node metastasis. This raises the question under which conditions a properly executed radical prostatectomy in high risk prostate cancer might provide a better course of action? Systematic review has shown that high risk prostate cancer men can be successfully treated with radical prostatectomy. However radical prostatectomy can not be considered as the monotherapy in this cohort of patients, but as a part of multimodal treatment, which can include radiation, hormonal and chemotherapies. PMID- 17404434 TI - Mortality differences between hemodialysis and peritoneal dialysis among ESRD patients in Georgia. AB - Comparisons of mortality rates in patients with end-stage renal disease (ESRD) on peritoneal dialysis and hemodialysis have been inconsistent. The aim of the study was to reveal the possible effect of comorbidity on differential survival in these two groups. 210 hemodialysis and 95 peritoneal dialysis patients were enrolled in this study who underwent dialysis treatment in the nephrology department of national centre of urology in Tbilisi between January 2004 and December 2006. Peritoneal dialysis was used by 9% of patents as baseline, 21% at twelve months and 26.5% at twenty four months. The mean follow-up was 24 months, with a maximum follow-up time of 36 months. The total co-morbidity was higher in hemodialysis compared to peritoneal dialysis patients at baseline, 12 months and 24 months. The mortality was similar in the groups defined by the mode of dialysis. Based on the results of the study, it is to conclude that, hemodialysis and peritoneal dialysis, as two different modalities of renal replacement therapy in the patients with ESRD, are associated with similar overall survival rates in Georgia. The apparent survival advantage of peritoneal dialysis may be due to lower co-morbidity and a lower burden of acute onset ESRD at the inception of dialysis. PMID- 17404435 TI - Dynamics of the protein spectrum changes in blood erythrocytes of male patients with prostate adenocarcinoma after plastic orchiectomy. AB - We have studied erythrocytes electrophoresis profiles in patients with prostate adenocarcinoma before and after plastic orchectomy and in healthy men to detect changes in the protein spectrum of erythrocytes. Our investigations have shown that during this pathology protein spectrum of erythrocytes structure undergoes substantial changes. This is demonstrated by losing of 35 KD molecular weight protein fraction and by appearance of 36 KD molecular weight protein fraction on the erythrocytes electrophoregram. It should be noted, that after plastic orchectomy we can see the relative normalization of erythrocyte's electrophoresis picture and it's approaching to the data of the control group (losing of 120 KD protein fraction and appearance of 48 KD protein fraction), which indicates to the normalization of the condition of the whole organism. It was detected that the synthesis of lower molecular protein fractions, on the one hand is influenced by rising of accantocytes percentage and on the other hand by the activation of the lipid oxidation process in erythrocytes membrane. PMID- 17404436 TI - Down-regulation of p27(Kip 1) cyclin-dependent kinase inhibitor in prostate cancer: distinct expression in various prostate cells associating with tumor stage and grades. AB - The goal of the study was to analyze the expression of p27(Kip 1) cyclin dependent kinase inhibitor protein (CDKI) in different cells of benign, malignant and hormonally treated prostate cancer tissue and assess their possible association with different clinical parameters. Expression of p27(Kip 1) CDKI was evaluated and compared in: 32 BPH, 20 prostate cancer (PCa) and 6 hormonally treated prostate cancer (HTPCa) tissues. Intensity of the expression was compared between the groups and association was soughed with the cancer clinical parameters. Total expression of p27(Kip 1) was significantly higher in BPH as compared with PCa (p=0.0002) and HTPCa (p=0.0324). The difference between PCa and HTPCa was not significant. p27(Kip 1) was higher expressed by epithelial, ductal and vascular prostatic cells of BPH as compared with PCa (p=0.0001, 0.0101 and 0.0224, respectively). The stromal expression of the marker was not different between the groups. Epithelial marker expression was significantly increased in HTPCa as compared with PCa (p=0.0460). In the PCa group, the intensity of the protein expression was negatively associated with the tumor stage, Gleason scores 1, 2, and the Gleason sum (p=0.0453, 0.0202, 0.0074 and 0.0098, respectively). This difference was found in epithelial, vascular and ductal prostatic cells. Down-regulation of p27(Kip 1) CDKI in PCa is detected in epithelial, vascular and ductal, but not the stromal cells. The intensity of the expression in these cells is associated with tumor stage and grades. The hormonotherapy is causing up regulation of p27(Kip 1) expression in prostate adenocarcinoma cells. PMID- 17404437 TI - Prevalence of urinary incontinence in women population. AB - Urinary incontinence among women population is widely spread disease, it occurs in every 5th, 6th woman. The frequency of disease is rising with age. Especially high percentage of disorder occurs in older respondents (60 years and >). Urinary incontinence is not only a medical, but also a social problem. Therefore, this disease is very actual problem and needs earlier revealing and exact diagnostic for timely and correct treatment. Data of our epidemiological researches corresponds to the foreign reference data. PMID- 17404438 TI - Posterior vertical lumbotomy, a clinical case. AB - From 1986 till February 2007 in National Center of Urology 444 patients were operated (249 women and 195 men, mean age 48.6 years) with upper urinary tract diseases. For access to the kidney and upper urether posterior vertical lumbotomy approach was performed. 27 patients were operated bilaterally. Total 471 operations were done by this approach. The results of operations with Posterior vertical lumbotomy were compared with flank incision for a variety of parameters (mean operative time, number of doses of postoperative parenteral analgesics, duration of postoperative hospitalization, wound infection, incisional hernia). Comparative analysis show, that posterior vertical lumbotomy is much less traumatic incision. PMID- 17404439 TI - Compartment syndrome, rhabdomyolysis and risk of acute renal failure as complications of the urological surgery. AB - Current knowledge related to the risk of CS when operating in these positions (Lloyd Davies and Trendelenburg tilt) is such that it can be deemed negligent to keep patients in this position (with legs higher than the heart) when not absolutely necessary. If it means repositioning and redraping, thus adding a few extra minutes to the episode and costing a small amount in additional drapes, surely this is a small price to pay for excellent perioperative care? If repositioning is impossible to execute, the head-down tilt position should be reversed every two hours, for a short period of time, to allow more natural perfusion of the lower limbs to occur. Raza et al recommend that if the anticipated procedure duration is beyond four hours, the legs should be removed from supports every two hours for a short period to prevent reperfusion injury. The use of Allen stirrups is preferred to calf supports or metal skids. Turnbull and Mills suggest that we should certainly review our use of compression stockings and intermittent compression devices when operating on patients in the Lloyd Davies position. It will be deemed negligent to misdiagnose (ie: mistake for a DVT) or delay treatment (by prolonged re-assessment) of CS postoperatively when patients have been subjected to prolonged surgery in these abnormal positions. Delayed or missed diagnosis may not only be limb-threatening (and cause a very protracted hospital stay)--it can be life-threatening. With today's current knowledge, surgeons undertaking prolonged surgery in abnormal positions must be aware of this, fortunately rare, complication. Practice guidelines within perioperative care should reflect current knowledge and ensure that risk is minimized. Patients who take legal action if they have experienced this condition may be awarded substantial costs against negligence if lack of care can be proven or diagnosis has been delayed. PMID- 17404440 TI - [Kidney protein complexes that inhibit gene expression in the nuclei of homotypic cells]. AB - The comparative study of protein complexes (PCs) isolated by alcohol extraction from white rat and human nephrocytes (post-operational material) have been performed. It has been shown that PCs of white rat cells inhibit RNA synthesis in the nuclei of intact as well as nephrectomized (unilateral nephrectomy) animals. The same effect has been shown while studying the PCs of patients with hydronephrotic kidneys. At the same time the inhibition of transcription has not been detected in case of RCC kidney cancer cell PCs. The chromatography studies revealed that the active component of PCs (12 kDa protein factor) which had been detected earlier has not been shown up in cancer cells of human kidney. PMID- 17404441 TI - [Some aspects of transurethral resection of large benign prostatic hyperplasia]. AB - Benign prostatic hyperplasia (BPH) is a progressive disease that affects the quality rather than the quantity of life of men. There are two methods of surgical treatment of BPH: transurethral resection of the prostate (TURP) and open prostatectomy. In this study we investigated the safety and efficacy of TURP for large prostate glands. To analyze our clinical data we can conclude introduction of technological innovations, especially of "low pressure" TURP, have made it possible to perform TURP for large prostates so safely as for recommended volumes. Also our trial has demonstrated that complications after TURP were within admissible limits of standard TURP. Therefore, TURP can be considered as an effective and safe procedure for patients with large prostate glands. PMID- 17404442 TI - [Radical perineal prostatectomy--surgical technique, first results]. AB - Prostate cancer is the most common malignancy in men and the second leading cause of cancer death. Radical prostatectomy is the most effective treatment for localized prostate cancer. With increasing use of minimally invasive treatment methods, clinical outcomes are becoming important assessment tools to compare one option to another. Perineal prostatectomy is modified to incorporate contemporary surgical ideas, including preservation of cavernosal nerve bundles, sphincteric urethra at the prostatic apex, and the bladder neck. During 2001-2004 in National Centre of Urology radical perineal prostatectomy was performed in 9 patients. The mean age of the patients was 64 years (range: 53-71 years). All operations were carried out with curative purpose for the treatment of localized prostate cancer. In all cases prostate specific antigen (PSA) was <10 ng/ml, Gleason score<7. There were analyzed operative time, volume of blood transfusions, duration of hospital stay, peri-operative complications, pre and postoperative potency and urinary continence. The distribution of pT categories was: pT2a-2 (22.2%); pT2b-5 (55.6%); pT3a-1 (11.1%); pT3b-1 (11.1%). The mean operative time was 130 minutes (range 95-180 minutes). The middling volume of blood transfusion per patient was 427 ml. At 12 months incontinence was manifested in 1 (11.1%) patient. Postoperative potency was reached in one case. Margins were positive in 2 (22.2%) cases. In both cases was manifested biochemical relapse. One of these patients died after 32 month from surgery. According our results 3-year tumor specific survival was in 7 (77.8%) patients. Radical perineal prostatectomy is an excellent alternative approach for radical surgery in the treatment of early prostate cancer. This method of prostatectomy is able to achieve complete cancer resection while preserving urinary and sexual function in the majority of men presenting with clinically localized prostate cancer. The simplicity and minimally invasive nature of this procedure contribute to a short recovery and low overall cost of therapy. The radical perineal prostatectomy is a cost efficient, outcome-effective minimally invasive method of treating men with localized prostate cancer. PMID- 17404443 TI - [The efficacy of extracorporal shock wave lithotrypsy in urinary stone disease]. AB - Extracorporal Shock Wave Lithotrypsy (ESWL) is "golden" standard therapy among the various methods of treatment of urinary stone disease. We have evaluated 1645 patients with urinary stone disease who underwent ESWL from 2003 to 2006, with third generation Dornier Compact Delta lythotriptor. 932 (56.7%) of them were male and 713 (43.3%) were female, 55 (3.3%) of all were children. Patients' age varied from 1 to 93 years. The total amount of sessions performed were 3391 (mean 2.1). In case of renal calculi the amount of shocks was 2000-3500 and the duration of session varied from 20 to 35 minutes. In case of urethral calculi- 3000-4500 shocks, duration of session varied from 30 to 45 minutes. From 1719 stones 1452 (84.5%) were X-Ray positive and 267 (15.5%) were X-Ray negative. ESWL was successful (stone free) in 1211 (73.6%) and semi successful in 91 (5.5%) cases; Semi successful group included patients, which became disobstructed after ESWL, but the some of the stone fragments were dislocated into lower calyx and were not eliminated. Unfinished treatment (Patient disappeared after the initial sessions) were in 262 (15.9%) cases. 81 cases were not successful. 31 patients from this group underwent urethrorenoscopy, 49 patients--open surgery and 7--PNL. In conclusion, ESWL is the "golden" standard treatment of Urinary stone disease in cases of carefully performed examination and management. Careful study of every single case, gives as the possibility to avoid expensive and invasive procedures and reduce the risk of complications. PMID- 17404444 TI - [Quantitative changes of testosterone and prostate-specific antigen at low and high densities of PSA in men with prostate cancer]. AB - In 61 patients with prostate cancer quantitative changes of testosterone and prostate-specific antigen (PSA) in blood serum at low and high densities of PSA have been investigated. It was shown that a high coefficient of correlation has been established in patients with prostate cancer and low density of PSA, but weak correlation was documented for the patients with prostate cancer and high density of PSA. PMID- 17404445 TI - [Using plaferon in patients with nephrolithiasis of the only kidney]. AB - We have studied the influence of the Plaferon on the stimulation of restoration of a kidney function in patients with nephrolithiasis of the only kidney. The researches were performed in 42 patients with nephrolithiasis of the only kidney (control group-23; group accepted Plaferon-19 patients). In group of the patients, which accepted Plaferon, the creatinine clearance after two weeks from the operation was increased (58.1+/-5.9 ml/min) and achieve statistically main differs from those in control group (40.1+/-4.2 ml/min; p<0.05). There was detected correlation between "osmolar free" water reabsorption in each nephron and mass of the active nephrons. In control group these correlations are weak and couldn't be evaluated as positive one. In both groups we couldn't detect correlation between excretion fraction of filtered Natrium and reabsorption of "osmolar free" water. Plaferon positively influences process of stimulation of restoration of a renal function at post-operative period in patients with nephrolithiasis of the only kidney and to restoration of the urine passage. After using Plaferon at post-operative period we can detect the better function of the glomerular filtration and other investigated parameters, which differs from those in control group. Plaferon should be used in patients with nephrolithiasis of the only kidney for stimulation of restoration of a renal function at postoperative period. PMID- 17404446 TI - [Prostate-specific antigen in serum and prostate epithelium in benign prostatic hyperplasia]. AB - The aim of our investigation was the simultaneous study of concentration of prostate-specific antigen (PSA) in serum and its immune-histochemical expression in the adenomatous prostatic tissue. 50 patients have been investigated. In 48 patients benign prostatic hyperplasia (BPH) has been diagnosed, and 2 patients served as controls. Investigated patients were divided by 2 groups: 1st group- patients with moderate IPSS (n=6); 2nd group--patients with expressed IPSS (n=42). The results of our investigations shown that at any variant of BPH lumine epithelium, as a rule, were PSA-positive. Negativity of immune-histochemical reaction was observed in areas of epithelium atypical hyperplasia, when proliferated epithelium is presented by basal cells. Expression degree varied between weak, moderate and high. The comparison of PSA expression degrees in tissue and its blood levels shown that the change of blood concentrations was more variable, than expression in tissue. Quantitative imbalance between them indicates that determination of blood PSA without immune-histochemical investigation is less reliable and does not give a valuable insight about the essence prostate pathology. PMID- 17404447 TI - [Cases of Leydig cell tumor in male infertility]. AB - 1745 patients at the Department of Urology of Tenon Hospital (Paris, France) from 1990 to 2006 and at the department of urology of Tbilisi State Medical University (Georgia) during last several months have been examined and counseled on male infertility. Leydig cell tumor was found in 4 patients, among them 3 by palpation and testicle echography, fourth patient (at the age of 33) with bilateral varicocele (III stage at the left, II stage at the right) was more interesting for us. Leydig cell tumor was found out at the scrotum exploration. Oligoteratozoospermia (OATS) has been distinguished from his spermogram. Microsurgical bilateral correction of varicocele and scrotum exploration, and double-sided double biopsy of testicle have been carried out. During examination of the left testis the hardly palpable node has been found out in the lower pole. We became compelled to make enlarged incision of tunica alba of the testicle lower pole. After that it was found out the solid, well encapsulated and yellow brown node (8 mm in diameter). The node enucleation away from tumor by 0.5 cm has been carried out. Exact histological investigation confirmed the presence of Leydig cell tumor. In 6 years after surgical operation the tumor node 1.8 mm in diameter has been found out in right (contra lateral) testis. The patient was urgently operated, ex-tempo investigation confirmed the presence of Leydig cell tumor in right testicle--high orchidectomy at the right has been carried out. Now, patient has not any symptoms of disease during 3.5 years of observation. Given case shows that the enucleation of Leydig cell tumor proves to be equivalent alternative of orchidectomy, which is suggested by many authors. Taking into account the presence of encapsulation and tumor benignity it is important to have opportunity of the testicle preservation due to infertility problems. However, this tactics must be carried out under strong observation due to the opportunity of relapse even in several days after surgical operation. PMID- 17404448 TI - [Rare case of benign urachus tumor during pregnancy]. AB - We described rare case of urachus fibromyoma at the 11th week of pregnancy. Urachus tumor was occasionally revealed during ultrasonographic examination. It was shown in previous studies that 90% of urachus tumors are malignant, so was decided on artificial abortion, by the method of manual vacuum aspiration. After the abortion patient was sent to the urologic clinic, and after biopsy was revealed diagnosis of fibromyoma of urachus. It was performed operation of partial cystectomy with excision of urachus tissue up to umbilicus. The decision of artificial abortion performance in these cases is disputable. PMID- 17404450 TI - Front-illuminated versus back-illuminated photon-counting CCD-based gamma camera: important consequences for spatial resolution and energy resolution. AB - Charge-coupled devices (CCDs) coupled to scintillation crystals can be used for high-resolution imaging with x-rays and gamma rays. When the CCD images can be read out fast enough, the energy and interaction position of individual gamma quanta can be estimated by a real-time image analysis of the scintillation light flashes ('photon-counting mode'). The electron-multiplying CCD (EMCCD) is well suited for fast read out, since even at high frame rates it has extremely low read-out noise. Back-illuminated (BI) EMCCDs have much higher quantum efficiency than front-illuminated (FI) EMCCDs. Here we compare the spatial and energy resolution of gamma cameras based on FI and BI EMCCDs. The CCDs are coupled to a 1000 microm thick columnar CsI(Tl) crystal for the purpose of Tc-99m and I-125 imaging. Intrinsic spatial resolutions of 44 microm for I-125 and 49 microm for Tc-99m were obtained when using a BI EMCCD, which is an improvement by a factor of about 1.2-2 over the FI EMCCD. Furthermore, in the energy spectrum of the BI EMCCD, the I-125 signal could be clearly separated from the background noise, which was not the case for the FI EMCCD. The energy resolution of a BI EMCCD for Tc-99m was estimated to be approximately 36 keV, full width at half maximum, at 141 keV. The excellent results for the BI EMCCD encouraged us to investigate the cooling requirements for our setup. We have found that for the BI EMCCD, the spatial and energy resolution, as well as image noise, remained stable over a range of temperatures from -50 degrees C to -15 degrees C. This is a significant advantage over the FI EMCCD, which suffered from loss of spatial and especially energy resolution at temperatures as low as -40 degrees C. We conclude that the use of BI EMCCDs may significantly improve the imaging capabilities and the cost efficiency of CCD-based high-resolution gamma cameras. PMID- 17404451 TI - Design of an endoscopic optical reference to be used for autofluorescence bronchoscopy with a commercially available diagnostic autofluorescence endoscopy (DAFE) system. AB - We present the design of a sterilizable optical reference to characterize and quantify the inter-patient variations in tissue autofluorescence during autofluorescence bronchoscopy with Richard Wolf's diagnostic autofluorescence endoscopy (DAFE) system. The reference was designed to have optical and spectral properties similar to those of the human bronchial wall in spectral conditions corresponding to autofluorescence bronchoscopy conducted with the DAFE system (fluorescence excitation at 390-470 nm and red backscattering light at 590-680 nm). The reference's effective attenuation coefficient and reflectance were measured at 675 nm. In addition, its fluorescence emission spectrum was determined under 430 nm wavelength excitation. The reference is photostable, reproducible, biocompatible and small enough to be easily inserted through the working channel of a conventional bronchofibrescope. This cylindrical (length: 2 mm; diameter: 2 mm) optical reference was validated in a clinical environment. PMID- 17404452 TI - Magnetization evolution in balanced steady-state free precession with continuously moving table. AB - Diagnostic imaging of systemic disorders, such as peripheral vascular diseases, requires a field-of-view (FOV) larger than the local FOV available on clinical MR scanners. The continuously moving table (CMT) method acquires large FOV images in a single acquisition. Balanced steady-state free precession (bSSFP) is an attractive candidate for the CMT method due to its short repetition time and high signal-to-noise ratio. However, introducing table motion during data acquisition perturbs the magnetization evolution towards steady state. In this paper, a computer model was developed to simulate the bSSFP magnetization evolution in the presence of table motion. From these simulations, predictions were made about the maximum table velocities that would allow the magnetizations of specific tissues to evolve to the theoretical steady-state values. These predicted maximum table velocities were then successfully verified in vivo with bSSFP CMT acquisitions. For an imaging FOV 400 cell cycle-regulated genes in normal human fibroblasts based on the proportions of cells in each phase of the cell cycle. By isolating effects on the gene expression associated with the cell cycle phase redistribution after genotoxin treatment, the direct transcriptional target genes were distinguished from genes for which expression changed secondary to cell synchronization. Application of this model to ionizing radiation (IR)-treated normal human fibroblasts identified 150 of 406 cycle-regulated genes as putative direct transcriptional targets of IR induced DNA damage. Changes in expression of these genes after IR treatment derived from both direct transcriptional regulation and cell cycle synchronization. PMID- 17404514 TI - Progressive and inhibitory cell cycle proteins act simultaneously to regulate neurotrophin-mediated proliferation and maturation of neuronal precursors. AB - Neuronal stem cell expansion and differentiation is a process involving stages of proliferation and maturation governed by the sequential and combinatorial exposure of cells to extrinsic factors. The olfactory epithelium is an excellent model to investigate regulation of this process, as it undergoes neuronal replacement post-natally. We have shown that the neurotrophins NGF and BDNF sequentially promote proliferation of developing olfactory sensory neuronal precursors, although their kinetics of proliferation and cell fate outcomes differ. Interestingly, CNP inhibits this neurotrophin-induced proliferation and promotes the maturation of these precursors to their next developmental stage. Here, we investigate the mechanisms behind these actions. Both NGF and BDNF increase the expression of cyclin D1 and cyclin-dependent kinase 4 (cdk4), with temporal expression patterns that parallel the proliferation kinetics of their cellular targets. The timing of cyclin D1 expression reflects differences in the need for transcription and translation in early and late stage precursors. CNP inhibits neurotrophin-induced cyclin D1 expression, and induces the expression of different profiles of inhibitory cell cycle proteins, which are neurotrophin specific and correlate with the attainment of different maturational cell fates. Inhibition of protein degradation reverses the effects of neurotrophins and CNP on cyclin D1 and inhibitor expression levels, respectively. These results suggest a model for cell cycle regulation that involves the simultaneous expression of progressive and inhibitory cell cycle regulatory proteins in response to both proliferation and differentiation agents, followed by selective degradation of these proteins, providing a mechanism for rapid and exquisite control of the cell cycle. PMID- 17404515 TI - The immunogenicity and reactogenicity of a new DTPw-HBV vaccine as a primary and booster vaccination course in healthy infants. AB - The immunogenicity and reactogenicity of primary vaccination at 3, 4 and 5 months and boosting at 12-18 months with a new DTPw-HBV vaccine was compared with either licensed DTPw and HBV vaccines given separately or a licensed DTPw-HBV combination (Tritanrix-HepB) in this randomized, partially double-blind primary vaccination and single-blind booster vaccination study in healthy infants (n = 239; Trial DTPw-HBV-001/004). One month after primary vaccination with the new DTPw-HBV vaccine, seroprotection against diphtheria, tetanus, hepatitis B and vaccine response to B. pertussis was seen in 100%, 98.7%, 94.9% and 98.7% of subjects, respectively, compared to 100%, > or =98.5%, 89.2% and 92.2% of subjects in the comparator groups, respectively. One month after the booster dose, a marked response to all vaccine antigens was observed, resulting in seroprotection against diphtheria, tetanus, hepatitis B in all DTPw-HBV recipients and response to B. pertussis in over 98.6%. After primary vaccination, there was evidence that fever > or =38.0 degrees C (rectal route) occurred more frequently after the new vaccine (following 41.6% of doses, compared with 32.2% and 29.3% in the comparator groups, p < 0.05) and that pain and drowsiness occurred more frequently than after licensed DTPw-HBV (45.3% versus 35.1% and 37.1% versus 24.9%, respectively). However after primary and booster doses Grade 3 symptoms occurred at similar frequencies in the three groups suggesting these possible differences are of minimal clinical significance. In conclusion, within the framework of this study the immunogenicity and safety profiles of GSK Biologicals' new DTPw-HBV vaccine when used for primary and booster vaccination were acceptable. PMID- 17404516 TI - [Neuromuscular complications in transplant recipients]. AB - Neuromuscular complications in transplant recipients are common and contribute to morbidity and mortality. Complications such as acute and chronic inflammatory demyelinating polyneuropathies and toxic myopathies are related to the changes in immune modulation that occur after transplantation or result from immunosuppressive treatment toxicity. Alternatively, other complications such as myositis, myasthenia gravis, and mononeuropathy multiplex may result from a dysimmune systemic disorder such as post-transplant lymphoproliferative disorder, graft-versus-host disease or hepatitis C virus or hepatitis B virus chronic infection. Lastly, some of these complications, e.g., compression or stretch of individual nerves or plexus, are commonly seen in a postoperative setting and are not specific of patients with organ transplantation. This review focuses on the characteristic features, management, prognosis and pathophysiology of common and immune-related neuromuscular complications in organ transplant recipients. PMID- 17404517 TI - [Indications for cerebral MR proton spectroscopy in 2007]. AB - Magnetic resonance spectroscopy (MRS) is being increasingly performed alongside the more conventional MRI sequences in the exploration of neurological disorders. It is however important to clearly differentiate its clinical applications aiming at improving the differential diagnosis or the prognostic evaluation of the patient, from the research protocols, when MRS can contribute to a better understanding of the pathophysiology of the disease or to the evaluation of new treatments. The most important applications in clinical practice are intracranial space occupying lesions (especially the positive diagnosis of intracranial abscesses and gliomatosis cerebri and the differential diagnosis between edema and tumor infiltration), alcoholic, hepatic, and HIV-related encephalopathies and the exploration of metabolic diseases. Among the research applications, MRS is widely used in multiple sclerosis, ischemia and brain injury, epilepsy and neuro degenerative diseases. PMID- 17404518 TI - [Unilateral spatial neglect: a dramatic but often neglected consequence of right hemisphere damage]. AB - INTRODUCTION: Unilateral Spatial Neglect (USN) is a common consequence of right brain damage. In the most severe cases, behavioral signs of USN can last several years and compromise patients' autonomy and social rehabilitation. These clinical facts stress the need for reliable procedures of diagnosis and rehabilitation. STATE OF THE ART: The last 3 decades have witnessed an explosion of studies on USN, which raises issues related to complex cognitive activities such as mental representation, spatial attention and consciousness. USN is probably a heterogeneous syndrome, but some of its underlying mechanisms might be understood as an association of disorders of spatial attention. A bias of automatic orienting towards right-sided objects seems typical of left USN. Afterwards, patients find it difficult to disengage their attention in order to explore the rest of the visual scene. Neglected objects are sometimes processed in an "implicit" way. PERSPECTIVES: The development of behavioural paradigms and of neuroimaging techniques and their application to the study of USN has advanced our understanding of the functional mechanisms of attention and spatial awareness, as well as of their neural bases. A number of new procedures for rehabilitation have recently been proposed. CONCLUSION: The present review describes the clinical presentation of USN, its anatomical basis and some of possible accounts of different aspects of neglect behavior. Results of computer simulations and of rehabilitation techniques are also presented with implications for the functioning of normal neurocognitive systems. PMID- 17404519 TI - [Standardization of the 5-word test in a group of 191 normal subjects aged 50 to 90 years]. AB - INTRODUCTION: The 5-word test (5WT) is a serial verbal memory test with semantic cuing. It allows to estimate cued learning, free recall, and cued recall of 5 words during immediate and delayed recalls (5 min later). The 5WT is proposed to rapidly evaluate memory of aging people. It has shown its sensitivity and its specificity in identifying patients with Alzheimer's disease (AD). OBSERVATIONS: We report the constitution of a sample of 191 French speaking normal subjects, aged from 50 to 90 years, in four age classes (50 years, 60 years, 70 years, 80 years) and three education levels. Total Score, Total Weighted Score, Free Delayed Recall, Delayed Recalls Total (memory score), and Free Recalls Total appear to be the best scores to appreciate the memory performance of the normal subjects. A Total Score of 10 was obtained in 74.9 p.cent of the subjects. No immediate nor delayed intrusions were recorded. In spite of the absence of an explicit consign, the recalled words were often ordered as they were in the list. CONCLUSION: The 5WT is a simple and reliable test for investigating memory in elderly people above 50 years old. PMID- 17404520 TI - [Mills' syndrome: a rare clinical entity]. AB - INTRODUCTION: Mills' syndrome is a rare motor neuron disease, initially described by Mills in 1900 as a progressive ascending or descending hemiplegia without significant sensory involvement. This syndrome is of uncertain nosological status, and is supposedly due to unilateral primary degeneration of corticospinal pathway. Some authors have suggested that it could represent a variant of primary lateral sclerosis. METHODS: We retrospectively studied the clinical and paraclinical data from eight patients with suspected Mills' syndrome hospitalized for diagnosis. RESULTS: For all patients, the clinical course was slowly progressive, with motor deficiency, unilateral pyramidal signs (or bilateral with asymmetry), without bulbar signs, fasciculations or sensory deficit. Final diagnosis was Mills' syndrome (n=3), primary lateral sclerosis (n=1), myelitis of unknown origin (n=2), progressive primary multiple sclerosis (n=1), and antiphospholipid syndrome (n=1). The main arguments for final diagnosis were brought by electrophysiology and brain and spinal MRI. CONCLUSION: Mills' syndrome is a rare clinical diagnosis, requiring exhaustive investigations. PMID- 17404521 TI - [Fatigue in neurological disease: different patterns in stroke and multiple sclerosis]. AB - INTRODUCTION: Fatigue is a complex, subjective experience, frequent in multiple sclerosis (MS) and stroke patients. The tiredness these patients experience can take on many features depending not only on the cerebral location of the lesions and mood aspects, but also on the pathophysiology of the disease. Thus, it is reasonable to expect that fatigue may have different implications in MS and stroke. The aim of the present work was to compare fatigue syndrome in these two populations. Patients were matched for handicap. MATERIALS AND METHODS: Seventy nine stroke and 39 MS outpatients were included with the following inclusion criteria: i) patients with possible or relapsing-remitting MS with an Expanded Disability Status Scale (EDSS) score<2.5, disease duration<6 years, and stable medical condition for at least 6 weeks; ii) stroke patients with mild neurological impairment, i.e. scoring<3 at the National Institute of Health Stroke Scale (NIHSS) one year after stroke; iii) absence of functional impairment (Barthel index=100) and similar negligible handicap (Rankin scale<2 for both groups); no or mild cognitive deficit; iv) neither DSMIV criteria of depression, nor significant anxious/depressive symptoms (Hospital Anxiety and Depression scale; HAD; score<8) in both groups. The Fatigue Assessing Instrument (FAI) was used to assess fatigue. RESULTS: Twenty-nine percent of stroke and 46 p. cent of MS patients had a significant score on the FAI (p<0.05). Multiple regression analysis using groups, gender and age as factors showed a group effect in 3 out of 4 subscales: MS patients scored higher than stroke patients mainly for psychic impact (4.86 vs. 3.28), but also for severity (mean 3.86 vs. 2.97) and specificity (4.36 vs. 3.32). Response to rest (5.36 vs. 6.06) only tended to be better in the stroke group. In the subpopulation with significant fatigue scores, psychic impact was more elevated in the MS group. The functional consequence of fatigue in physical, professional and social activities were similar. DISCUSSION: Fatigue was more severe in MS than stroke patients, independently of disability. The most significant factor in the MS group was the psychic impact, reflecting impaired motivation, concentration and irritability, despite the absence of depression. However, subjective consequences of fatigue on work, family and leisure activities were comparable in both groups. PMID- 17404522 TI - [Intravenous rt-PA for acute ischemic stroke: 69 consecutive patients managed in an emergency stroke centre]. AB - INTRODUCTION: After the 2002 European agreement on the use of rt-PA for fibrinolysis within less than 3 hours after ischemic stroke, we designed a specific patient management scheme for patients referred to our center. METHODS: We report the activity of the "stroke emergency" pathway in the Purpan Hospital (Toulouse) for 4 years. We wanted to evaluate our daily practice and to confirm that the results obtained in the randomized clinical trials with rt-PA can be reproduced in routine practice. RESULTS: Among all stroke patients treated in the Neurology Department, 10.2 per cent were managed via this new pathway, in order to receive a fibrinolytic treatment. Amongst these, 25.6 per cent were treated with rt-PA (2.6 per cent of all ischemic and hemorrhagic strokes, with an average NIHSS score of 15.8 at admission [5; 25]. In 90 per cent of the cases, potential patients for thrombolysis were selected by CT-scan. Time from onset to treatment averaged 2 h 25 min, whilst door-to-treatment time averaged 40 minutes. Two patients (3 percent) showed a symptomatic intra-cerebral hemorrhage. Death rate was 18.8 per cent. After 3 months, 53.5 per cent of patients were regarded as functionally "independent" (Rankin scale<3). CONCLUSION: These results in our unit confirm the feasibility, reproducibility, efficacy and safety of the rt-PA fibrinolytic treatment for ischemic stroke of less than 3 hours. A "Stroke emergency" pathway appears to be a helpful option to treat as many patients as possible with the shortest possible lead times. PMID- 17404523 TI - [Adult leukoencephalopathy caused by alpha-mannosidosis deficiency]. AB - Adult leukoencephalopathy caused by alpha-mannosidosis deficiency (MIM248500) is a recessive inherited lysosomal storage disease associated with decreased activity of alpha-mannosidase. This enzyme degrades oligosaccharides and glycoproteins in neural and visceral tissues. There are two different disease phenotypes, type-I or severe infantile phenotype and type 2, which progresses more slowly and is compatible with survival into adulthood. We report the case of a 51-year-old man with gait disorders beginning at the age of 40 years associated with leukoencephalopathy due to alpha-mannosidosis deficiency. PMID- 17404524 TI - [Nitric-oxide triggered neurological disorders in subjects with vitamin B12 deficiency]. AB - INTRODUCTION: Nitrous oxide is frequently used for anesthesia. It may cause spinal cord toxicity. CASE REPORTS: We report two patients who presented gait disorders after nitrous oxide anesthesia. Physical examination revealed arms and legs pyramidal syndrome and abnormal proprioception, consistent with subacute combined degeneration of the spinal cord. Serum vitamin B12 level was extremely low. The patients improved with parenteral treatment with hydroxycobalamin. CONCLUSIONS: The inactivation of methionine synthase and L methylmalonylcoA mutase by nitrous oxide has been previously demonstrated. Anesthesia-related exposure to nitrous oxide may induce neurologic disorders even in patients with no preliminary vitamin B12 deficiency. PMID- 17404525 TI - [Brain magnetic resonance imaging and neuropathology of cortical laminar necrosis]. AB - INTRODUCTION: The most frequent acute and sub-acute complications of chronic alcoholism are delirium tremens, hepatic encephalopathy and Gayet-Wernicke encephalopathy. Morel laminar sclerosis is a rare and less known complication, often reported with Marchiafava-Bignami disease. CASE REPORT: A 57-year-old alcoholic man presented delirium after surgery. Anterograde and retrograde amnesia as well as wrong recognitions appeared progressively and one generalized seizure occurred. He then developed mutism and became bedridden. Magnetic resonance imaging (MRI) showed high-intensity bilateral temporoparietal signals from white matter on T2-weighted images and high-intensity signals from the parietal cortex on T1-weighted images. The patient died four months after the onset of the delirium. Post-mortem examination of the brain showed cortical laminar necrosis with Alzheimer Type II gliosis but without demyelinisation of the corpus callosum. CONCLUSION: Cortical laminar necrosis with chronic ethylism is usually called Morel's laminar sclerosis. Nevertheless, histology is not typical of this diagnosis, because of necrosis especially of the second (and not the third) layer of the cortex, and because of the absence of lesion of the corpus callosum. MRI data are of interest here because they were rarely reported in cases of Morel's laminar sclerosis. PMID- 17404526 TI - [One episode of meningoencephalitis, three diagnoses]. AB - Meningo-encephalitis is a set of threatening diseases. The treatment needs to be started quickly for pathogens such as herpes simplex virus type 1 or Listeria monocytogenes. Apart from these classical etiologies, many other diseases may induce meningo-encephalitis. We report the case of a patient, infected with HIV, who presented a history of meningo-encephalitis due to herpes simplex type 1. Three weeks later, he presented an encephalopathy due to aciclovir and then we discovered a chronic meningitis in relation with his HIV infection. PMID- 17404527 TI - [Therapeutic management of tics in Tourette's syndrome]. AB - Tourette's syndrome is a neuropsychiatric disorder characterised by both involuntary movements, tics, and psychiatric symptoms, attentional deficit disorder, hyperactivity, obsessive compulsive symptoms..., and can be the cause of major disability. Over the past 30 years, several types of treatment have been proposed for the treatment of tics in Tourette's Syndrome, ranging from psychotherapeutic approaches to neurosurgery. The education of the patient and his entourage is fundamental and must be offered to all patients. Psychotherapy does not directly improve the tics but contributes to a better adjustment of both patient and carers to his disability. The decision to start a course of drug therapy depends largely on the impact of the patient's tics on his personal life. Drug treatment relies on neuromodulators acting on a variety of neural systems and whose efficacy has been rarely demonstrated. The literature shows that the latest generation of dopaminergic antagonists have the highest benefit/risk ratio. Recently, deep brain stimulation, by modulating neuronal activity in structures involved in the pathophysiology of the disease, has become a promising therapeutical approach, producing a marked decrease in the severity of tics over that obtained with other treatments. PMID- 17404529 TI - [Editorial]. PMID- 17404528 TI - [Capillary telangiectasis]. PMID- 17404530 TI - [Lung cancer and oncogeriatrics - Current status]. PMID- 17404531 TI - [Contribution of echoendoscopy to mediastinal staging]. PMID- 17404532 TI - [Small-cell lung carcinoma: can we improve the results of the treatment by associating another therapy to the chemotherapy?]. PMID- 17404533 TI - [Recent advances in malignant pleural mesothelioma management]. PMID- 17404534 TI - [Treatment of stage III non-small lung carcinomas: is there controversy?]. PMID- 17404535 TI - [Treatment of unresectable stage III non-small-cell lung carcinomas]. PMID- 17404536 TI - [From ASCO 2006 to clinical practice]. PMID- 17404537 TI - [Chemotherapy duration in advanced-stage non-small-cell lung cancers]. PMID- 17404538 TI - [Second-line treatment of non-small-cell lung cancers]. PMID- 17404539 TI - [Is it necessary to select patients to prescribe targeted therapies in stage IV non-small-cell lung cancers?]. PMID- 17404540 TI - [Markers of thrombotic disease: procoagulant microparticles]. AB - Thrombosis remains one of the leading causes of mortality and morbidity in developed countries. Relevant markers of the primary thrombotic risk however remain of limited accessibility, and clinicians are left with markers of essentially etiological nature. Fortunately, new entities, testifying to cellular activation or damage within the vascular compartment, have been recently described and are in the validation process. Microparticles (MP) are plasma membrane fragments released by stimulated or apoptotic cells. In the vascular compartment, they constitute a disseminated storage pool of bioactive effectors involved in inflammation, thrombosis, vascular tone, angiogenesis. Their biological characteristics are predetermined by the cytosolic and membraneous components hijacked from the activated cells. Their procoagulant properties are based on, (i) the accessibility of phosphatidylserine, a procoagulant aminophospholipid exposed after stimulation and necessary for the assembly of the blood clotting enzyme complexes, and (ii) the possible presence of tissue factor, the major initiator of the coagulation cascade. The incidence of MP in haemostatic processes has been demonstrated in physiology and pathology. They are now considered true pathogenic markers of the thrombotic risk. PMID- 17404541 TI - [New antitoagulants]. AB - In contrast to older anticoagulant agents vitamin K antagonists and heparins, the new ones are directed towards a single target in general. The main characteristics of the new agents are: their site of action in the coagulation cascade and their mechanism of action which is indirect, antithrombin dependent, most often such as Fondaparinux and Idraparinux or direct such as Dabigatran, Rivaroxaban; the specificity of the new molecules, since they must not interact with other enzymes: trypsin, kallikrein, t-PA, etc...; their mode of administration parenteral and/or oral; their pharmacokinetics and their clearance frequently by the kidney (Hirudin, fondaparinux) or through hepatic metabolism (argatroban); tolerance including for all compounds the bleeding risk or an unexpected hepatic intolerance for Ximelagatran; the availability of a specific antidote and the cost of the drug; one compound is registered in France Arixtra Fondaparinux in major orthopedic surgery and in the treatment of venous thromboembolism and in prophylactic treatment in medical patients. However, the main indications of interest for these new drugs is atrial fibrillation. There is a real need in this indication and the number of patients to treat is growing with the longer life expectancy. PMID- 17404542 TI - [Presentation. Enzymes: the long pathway from genes to drugs]. PMID- 17404543 TI - [Discovery and crystallographic structure of human apolipoprotein]. AB - We report the serendipitous discovery of a human plasma phosphate binding protein (HPBP). This 38 kDa protein is co-purified with paraoxonase (PON1). The association between HPON1 and HPBP is modulated by phosphate and calcium concentrations. The HPBP X-ray structure solved at 1.9 A resolution is similar to the prokaryotic phosphate solute-binding proteins (SBPs) associated with ATP binding cassette transmembrane transporters, though phosphate-SBPs have never been characterized or predicted from nucleic acid databases in eukaryotes. However, HPBP belongs to the family of ubiquitous eukaryotic proteins named DING, meaning that phosphate-SBPs are also widespread in eukaryotes. The absence of complete genes for eukaryotic phosphate-SBP from databases is intriguing, but the astonishing 90% sequence conservation of genes between evolutionary distant species suggests that the corresponding proteins play an important function. HPBP is the first identified transporter capable of binding phosphate ions in human plasma. Thus it is thought to become a new predictor and a potential therapeutic agent for phosphate-related diseases such as atherosclerosis. PMID- 17404544 TI - [Exploring the conformational energy landscape of acetylcholinesterase by kinetic crystallography]. AB - Acetylcholinesterase is a very rapid enzyme, essential in the process of nerve impulse transmission at cholinergic synapses. It is the target of all currently approved anti-Alzheimer drugs and further progress in the modulation of its activity requires structural as well as dynamical information. Exploration of the conformational energy landscape of a protein by means of X-ray crystallography requires the use of experimental tricks, to overcome the inherently static nature of crystallographic structures. Here we report three experimental approaches that allowed to gain structural insight into the dynamics of acetylcholinesterase, which is relevant for structure-based drug design. PMID- 17404545 TI - [Enzyme optimization, mutagenesis and directed evolution]. AB - Among the different areas of biotechnology, enzyme engineering represents a growing field where major progress has been recently made. Indeed, chemical, pharmaceutical or food industries have increased needs for enzymes. This increase requires enzyme optimization in order to achieve, together or separately, greater operational stability, better specificity, increased solubility or preferential enantioselectivity. Directed and random mutagenesis, the classical methods of enzymatic engineering, have proved to be efficient in some cases, but are quite tricky. Directed evolution is a hybrid method recently developed in order to reproduce the random mechanisms of evolution in vitro. This method has now been used to optimise an increasing number of enzymes. In our research group, a directed evolution project has been initiated on a bacterial phosphotriesterase, a promising enzyme, capable of efficiently detoxifying organophosphorus nerve agents. PMID- 17404546 TI - [Substituted cyclodextrins: an example of biomimetic catalyzers]. AB - Among all molecules used to develop biomimetic catalysts, cyclodextrins are extremely attractive compounds. These oligosaccharides can form inclusion complexes with various organic substrates and in particular with organophosphorus poisons, which are widely used as chemical weapons and insecticides. Soman, a frightening neurotoxic agent, once "trapped" in the internal cavity of beta cyclodextrin can moreover undergo the nucleophilic attack of an oligosaccharide hydroxyl group, miming the first step of the enzymatic process. Selective substitution of beta-cyclodextrin by a 2-iodosobenzoic acid derivative has enabled effective synthesis of scavangers against organophosphorus compounds. Hydrolysis trials were carried out with paraoxon, as an organophosphorus model. The OP-hydrolyzing activity could reach more than two order of magnitude compared with free 2-iodosobenzoic acid. Nevertheless, hydrolysis of paraoxon showed saturation kinetics. Although the activity was strongly dependent on the relative position of the reactive group, these results showed the interest of a strategy, resulting in the "trapping" of the organophosphorus substrate in the internal cavity of the oligosaccharide in order to maintain it near the catalytic function. PMID- 17404547 TI - [Liposomes: support for the formation of stable capsules made of reticulated polyelectrolytes or silicum]. AB - Uses of enzymes for therapeutic purpose or for biosensing require a well controlled nanoenvironnement to avoid degradation by proteolytic agents, pH variations or dilution effects. A solution is encapsulation under undenaturating conditions into a nanometer sized and stable capsule. The nanometer scall decreases recognition by the reticulo-endothelial system recognition and subsequent immune reaction. Liposomes are the method of choice since they allow protein encapsulation under mild conditions. However they lack in stability. In contrast, other type of capsules exhibit strong stability but with conditions required for formation that are incompatible with enzyme integrity. Here we combine different capsule formation techniques and use liposomes as templates for further stabilization. Here we demonstrate two types of multicomposite capsules. The first type is to coat the liposome surface with polyelectrolytes followed by secondary covalent crosslinking of the polyelectrolytes multilayer. In the second type of capsules we used silica to build an inorganic shell around liposome. Both techniques allow the formation of detergent stable nanocapsules which exhibits properties protective against acetylcholinesterase protein degradation, an enzyme of much interest for pesticide detection. PMID- 17404548 TI - [Topical enzyme-based formulations]. AB - Research studies aimed at improving the enzyme catalytic efficiency and operational stability are considered as a recurrent challenge with topical enzyme based formulations. A large number of plant and bacterial proteolytic enzymes have been studied as an early substitutive replacement for the mechanical debridement of partial-thickness burns. In addition, progress in protein engineering and vectorisation of drugs provide improvements in the approach of topical and photoprotective enzyme therapy. PMID- 17404557 TI - Noninvasive ventilation with positive airway pressure in paediatric intensive care. AB - AIM: The aim of the present study is to retrospectively evaluate the effectiveness of noninvasive pressure ventilation in the 24-bed Pediatric Intensive Care Unit (PICU) of the G. Gaslini Institute during a 24-month period. METHODS: A retrospective analysis of the characteristics (pH, CO2, SpO2, respiratory rate, oxygen requirement) of patients treated with noninvasive mechanical ventilation for different acute pathologies has been performed. RESULTS: Twenty patients (mean age 7.4+/-0.28 years) with acute respiratory failure due to different pathologies were treated with noninvasive mechanical ventilation. They were divided into 2 groups: the hypoxic group, suffering from pulmonary diseases, and the hypercapnic group, presenting a failure of the mechanical strength or increased dead space. Modalities of ventilation were pressure assisted/controlled or pressure support, delivered through nasal or facial masks. Fifteen out of 20 patients presented a marked improvement of oxygenation and ventilation. Mean times of treatment were 69 and 200 h in the hypoxic and hypercapnic groups, respectively. Five patients required intubation. Two patients presented reversible skin lesions over the nasal bridge. CONCLUSIONS: Noninvasive ventilation can be used in PICU. Major advantages regard immunocompromised children and patients with exacerbations from chronic respiratory diseases, whereas the exact role of noninvasive positive pressure ventilation in patients affected by acute respiratory distress syndrome is still controversial. PMID- 17404558 TI - A comparison of nasal intermittent versus continuous positive pressure delivery for the treatment of moderate respiratory syndrome in preterm infants. AB - AIM: Nasal continuous positive airway pressure and nasal intermittent positive pressure ventilation have both been proposed as a form of gentle respiratory support for neonatal respiratory distress syndrome. We have compared these two respiratory support methods in the management of early stages of mild-moderate, neonatal respiratory distress syndrome. METHODS: A prospective, randomized trial was performed enrolling 88 consecutive preterm infants with mild to moderate respiratory distress syndrome matched for birthweight, gestational age, sex, admission FiO2 and Apgar scores. Nasal continuous positive airway pressure was delivered at a pressure of 4-6 cmH2O. Infants on intermittent pressure were put on ventilatory rates of 40 breaths per minute. Peak pressures of 14-20 cmH2O and end-expiratory pressures of 4-6 cmH2O were used judging clinically the chest cage expansion. RESULTS: Although there were no differences in the PaO2 values between the groups, infants treated with nasal intermittent positive pressure ventilation showed a significantly lower pCO2 values than those in the continuous pressure group (40+/-2 vs 58+/-4 mmHg, P<0.05). Also a significantly lower incidence of apnoeic episodes (number of episodes/hour 0.4+/-0.2 vs 0.9+/-03; P<0.05) and a shorter respiratory support in the nasal intermittent positive pressure ventilation group were observed. No difference in severe abdominal distension or need of endotracheal intubation was noted. CONCLUSIONS: In the present clinical setting, the use of nasal positive airway pressure in an intermittent way is associated to a more physiological arterial carbon dioxide tension, less apnoea and a shorter duration of respiratory support when compared with continuous pressure delivery by the same route. PMID- 17404559 TI - [Pulmonary infections in preterm newborns]. AB - AIM: The aim of this study was to evaluate retrospectively the incidence and etiology of connatal pneumonia and ventilator-associated pneumonia (VAP) in preterm newborns who had birth weight = or < 1250 g and required intubation for at least 12 h. METHODS: We have reported data about preterm newborns who had birth weight = or < 1250 g and required intubation for at least 12 h with diagnosis of connatal pneumonia and VAP, admitted to the neonatal intensive care unit from 1994 to 2004. We divided these 11 years into 4 periods. For each period we determined etiology associated with connatal pneumonia or VAP. RESULTS: A total of 417 patients were studied; 311 (74.6%) required mechanical ventilation (MV) for more than 48 h (the least for the diagnosis of VAP). Connatal pneumonia occurred in 35/417 patients (8.4%). VAP incidence did not change over time showing a slight increase in the last 2 years (from 27% to 33%). Mycoplasma and Chlamydia as causative organisms of connatal pneumonia dissapear during years. Gram-negative micro-organisms were isolated more frequently in last years in VAP episodes. CONCLUSIONS: The incidence of VAP does not decrease over time although length of MV was reduced. Additional studies are needed to improve criteria for the diagnosis and prevention of VAP in NICU patients. PMID- 17404560 TI - Cardiac functions by myocardial performance index and QT dispersion in survivors of childhood lymphoblastic leukaemia. AB - AIM: Childhood leukaemia treatment contains multiple chemotherapeutic agents in high doses that can cause severe toxic effects on heart and other vital organs. In this respect patients taking cancer chemotherapy are followed for these adverse effects. Echocardiographic myocardial performance index (MPI) was reported as a new method of combined systolic and diastolic function for both adults and children, calculated as isovolumic relaxation time plus isovolumic contraction time divided by ejection time. In addition, it has been postulated that increased inter-lead differences in QT interval (QT dispersion) may be associated with an increased risk of cardiac death. The aim of this study was to determine the probable immediate and late adverse effects of childhood leukaemia treatment containing moderate dose of anthracyclines on heart by MPI and corrected QT dispersion (QTcD). METHODS: MPI and QTcD in 55 children with leukaemia and 38 healthy controls matched for age and sex were evaluated. RESULTS: There was no statistically significant difference between MPI values of patients and controls (20.7+/-13.1 (1-59.4) and 16.1+/-13.5 (0.3-77.5), P: 0.1, respectively). Also, there was no significant difference in MPI and QTc values between patients taking active treatment and those who completed the therapy and between the patients given a cumulative dose of anthracycline lower and higher than 250 mg/m2. But QTcD values were found to be higher in patients than controls (0.08+/-0.03 and 0.03+/-0.01, P<0.01, respectively). CONCLUSIONS: There was no overt cardiotoxicity in our children with leukaemia treated with protocols of ALL BFM 95 and TRALL 2000 (Modified BFM in Turkey) containing moderate dose of anthracyclines. However, they can cause subclinical cardiotoxicity and further monitoring and evaluation with such sensitive and noninvasive methods over a longer period of time are needed. PMID- 17404561 TI - Pediatricians' advices on timing of solid food introduction: two Italian experiences. AB - AIM: The aim of the study was to evaluate and compare methods of weaning suggested by pediatricians in two Italian regions with different geographical and cultural characteristics, Tuscany and Apulia. METHODS: Questionnaires were sent to 30 pediatricians, 15 in Tuscany and 15 in Apulia. Questions regarded prevalence and duration of breastfeeding, timing and manner of introducing other foods, and use of commercial baby food or food prepared at home. RESULTS: The pediatricians advised beginning weaning between the 4(th) and the 6th months. In Tuscany, 60% of pediatricians advised breastfeeding for 6 months and 40% for 12 months. In Apulia the same figures were 86.7% and 13.3%. All pediatricians advised differentiated introduction of foods. The percentages of Tuscany pediatricians suggesting late introduction of certain foods were 31.70% for eggs, 19.51% for cow's milk, fish and tomato, 4.87% for vegetables and 2.44% for citrus and gluten. Foods introduced with caution by Apulia pediatricians were egg white (41.37%), gluten (31.03%) and fish (27.58%). The percentages of Tuscany and of Apulia pediatricians recommending home cooked food were 77.7% and 31.21%, respectively; 22.3% and 68.75% respectively recommended commercial baby food. CONCLUSIONS: Pediatricians in both regions are aware of the importance of nutritional quality in the first year of life and provide indications about the timing and manner of introducing foods in line with international scientific guidelines. PMID- 17404562 TI - [Description and evaluation of creative thinking in preterm low birth weight infants]. AB - AIM: Since the 1950s, the problem of how to evaluate creativity has been addressed in studies on the definition of measurement criteria and on the relationship between intelligence and creative thinking. Many revealed cognitive and relational disorders in preterm infants, particularly in preterm very low birth weight infants (birth weight <1500 g) and in infants with serious complications. This study describes the development of creative thinking in a group of children born preterm. METHODS: The study sample was 43 children (21 males, 22 females; age range 6-11 years), regularly attending school, born with low birth weight (1050-2450 g) at 29-32 weeks gestational age, and compared with a control group with birth weight >2500 g. The test battery included: Torrance Test of Creative Thinking (TCTT); WISC-R intelligence test; Goodenough Human Figure Drawing Test. RESULTS: Statistical analysis (Mann-Whitney U test) showed a statistically significant difference (P>0.05) between the 2 groups; scores for figure originality, figure fluidity and figure elaboration were consistently higher in the control group. Within the low birth weight group, there was a significant correlation (Spearman r) between verbal IQ and verbal fluidity and verbal flexibility subscale scores and between IQ performance and figure elaboration. Scores on the figure drawing tests showed higher creative ability in the control group. CONCLUSIONS: In children born preterm with low birth weight, emotive dynamics and flow of affection may influence the channels of communication between child and family. The low figure originality subscale scores support the hypothesis that psychodynamic and relational factors (worry about the preterm condition, overprotective behaviour by parents and others) could lead to diminished autonomy, flexibility and manipulatory interest in the child. PMID- 17404563 TI - Osteoporosis in children with neuromuscular diseases and inborn errors of metabolism. AB - The effects on bone of cerebral palsy (CP), Duchenne's muscular dystrophy and different metabolic diseases are reviewed from the literature. Children affected with neuromuscular diseases and inborn errors of metabolism may develope osteoporosis. Mechanical stimulation is paramount for bone strengthening, and immobilization is a well-known cause of osteoporosis. CP is the most common cause of disability in pediatrics. The main cause of low bone density in children and adolescents with CP and muscular dystrophy is lack of activity, but nutritional issues and pharmacological treatments can contribute to the clinical picture. Programs to exert mechanical stimulation of their bones are warranted, as much as nutritional programs. Treatment with bisphosphonates shows promising results in this population. The term ''inborn errors of metabolism'' comprise a large list of defects in the metabolism of amino acid transport and metabolism of peptides, carbohydrates, vitamins, minerals, and fatty acids. Other disorders included are errors in mitochondrial energy metabolism, problems with biosynthesis and breakdown of complex molecules, and neurotransmitter defects. Low bone density and fractures in these patients may be consequence of immobilization and muscle weakness, but also of treatments (e.g. steroids, dietary restrictions), and the primary disease. Adequate control of the primary disease is paramount to prevent bone problems. PMID- 17404564 TI - Enterobacter sakazakii an emerging bacterial pathogen with implications for infant health. AB - Enterobacter sakazakii (E. sakazakii) is an opportunistic pathogen and the aetiological agent in rare but life-threatening cases of meningitis, necrotizing enterocolitis, and sepsis in infants. Among infants, those at greatest risk are neonates (<28 days), particularly those born prematurely or of low birth weight (<2500 g). Consumption of contaminated powdered infant formula (PIF) has been epidemiologically linked with cases of infection. Contamination can occur during the manufacturing process or during postmanufacture reconstitution of formula. Development of rapid, sensitive and specific detection methods will facilitate manufacturers efforts to reduce the occurrence of E. sakazakii in the final powdered product. Furthermore, since PIF is not a sterile product, proper precautions should be taken during handling and reconstitution of formula prior to feeding in order to prevent contamination and proliferation of the bacterium. PMID- 17404565 TI - [Spinocerebellar ataxias in infancy: pathogenesis of potassium and calcium channels' diseases, clinical features and therapeutical approach]. AB - In infancy, the autosomal dominant inherited ataxias are severe neurological diseases, due to inherited mutations of ion channels. The main forms are: episodic ataxia type 1 (EA1), episodic ataxia type 2 (EA2), spinocerebellar ataxia type 6 (SCA6). EA1 is due to a mutation in KCNA1, the gene encoding human Kv1.1 on chromosome 12p13, which contributes as a subunit to the formation of potassium channels in motor nerve terminals and in many central nervous system neurones. To date, there are fifteen different mutations, which affect potassium channel's properties and lead to phenotypic variability and to different responses to therapy. EA2 can result from mutations in the CACNA1A gene, encoding calcium channels on chromosome 19p13.1 and widely distributed throughout the central nervous system. To date, associated with EA2, in the CACNA1A gene thirty different mutations have been described, resulting in altered or truncated protein products and, as a consequence, in nonfunctional calcium channels. There is phenotypic variability, also inside the same family, without correlation genotype-phenotype. SCA6 is a progressive neurodegenerative disease due to mutations of the CACNA1A gene. CACNA1A is responsible for both EA2 and SCA6. Nevertheless, the pathogenesis of the two diseases is different: SCA6 is associated with small expansion of a CAGn repeat, while EA2 is due to point mutations. Clinically, SCA6 is characterized by a slowly progressive development and by an inverse correlation between the number of repeats and the severity of the disease. PMID- 17404566 TI - Neonatal adrenal hemorrhage presenting as contralateral scrotal ematoma. AB - Neonatal adrenal hemorrhage may rarely present as scrotal hematoma. This condition can simulate a torsion of the testis requiring an immediate surgical exploration of the scrotum. Ultrasonographic study of the abdomen can find out intraperitoneal or retroperitoneal bleeding in order to avoid unnecessary surgical intervention. The case of a newborn with right adrenal hemorrhage presenting as left scrotal hematoma is reported. PMID- 17404567 TI - [Prevention and security of children today: the virtual world and identity thieves]. PMID- 17404568 TI - Parafibromin tumor suppressor enhances cell growth in the cells expressing SV40 large T antigen. AB - Parafibromin (PF) is a 531-amino acid protein encoded by HRPT2, a putative tumor suppressor gene recently implicated in the autosomal-dominant hyperparathyroidism jaw tumor familial cancer syndrome and sporadic parathyroid carcinoma. To investigate effects of PF's overexpression on cell proliferation, we performed assays in four different cell lines. The transient overexpression of PF inhibited cell growth in HEK293 and NIH3T3 cells, but enhanced cell growth in the SV40 large T antigen-expressing cell lines such as 293FT and COS7 cells. In 293FT cells, PF was found to interact with SV40 large T antigen and its overexpression promoted entry into the S phase, implying that the interaction enhanced progression through the cell cycle. The tumor suppressor protein PF acts as a positive regulator of cell growth similar to an oncoprotein in the presence of SV40 large T antigen. PMID- 17404569 TI - Aberrant FoxM1B expression increases matrix metalloproteinase-2 transcription and enhances the invasion of glioma cells. AB - We recently showed that FoxM1 is overexpressed in human glioblastomas and that forced FoxM1B expression in anaplastic astrocytoma cells leads to the formation of highly invasive glioblastoma multiforme (GBM) in nude mice. However, the molecular mechanisms by which FoxM1 enhances glioma invasion are unknown. In this study, we found that FoxM1 overexpression increased matrix metalloproteinase (MMP)-2 expression in glioma cells, whereas blockade of FoxM1 expression suppressed MMP-2 expression. Transfection of FoxM1 into glioma cells directly activated the MMP-2 promoter, whereas inhibition of FoxM1 expression by FoxM1 siRNA suppressed its activation. We identified a FoxM1-binding site in the MMP-2 promoter and demonstrated that FoxM1 protein bound directly to it. Mutation of this FoxM1-binding site significantly attenuated MMP-2 promoter activity. Furthermore, FoxM1 overexpression increased the invasiveness of glioma cells, whereas inhibition of FoxM1 expression suppressed the invasiveness of GBM cells. Inhibition of MMP-2 by a specific MMP-2 inhibitor reversed the invasive phenotype of glioma cells overexpressing FoxM1. Finally, immunohistochemical analysis of 45 human GBM specimens showed a significant correlation between FoxM1 overexpression and elevated MMP-2 expression. Collectively, these findings provide evidence that FoxM1 contributes to glioma progression by enhancing MMP-2 gene transcription and thus tumor-cell invasion. PMID- 17404570 TI - p53 and Delta Np63 alpha differentially bind and regulate target genes involved in cell cycle arrest, DNA repair and apoptosis. AB - The mechanism by which the p53 family of proteins coordinately regulates select target genes after various types of cell stress is not well understood. To further define factors that dictate regulation of target genes, we examined the binding of p53, DeltaNp63alpha and RNA polymerase II (pol II) to the regulatory regions of select target genes in primary human epidermal keratinocytes (HEKs) using chromatin immunoprecipitation. In rapidly proliferating cells, we observed constitutive binding of DeltaNp63alpha and varying levels of p53 binding, to consensus sites in target genes involved in cell cycle arrest, DNA repair and apoptosis. Following genotoxic stress, p53 occupancy increased whereas DeltaNp63alpha occupancy decreased at the majority of binding sites examined. Microarray analysis of transcripts isolated from HEKs ectopically expressing p53 and DeltaNp63alpha revealed an inverse regulation of select target genes by the two family members. Collectively, our results suggest that DeltaNp63alpha can function as a repressor of select p53 target genes involved in growth arrest, DNA repair and apoptosis, and that the location of the p53 consensus binding site(s) in a target gene may dictate whether pol II is constitutively bound in proliferating cells. PMID- 17404571 TI - RASSF6 is a novel member of the RASSF family of tumor suppressors. AB - RASSF family proteins are tumor suppressors that are frequently downregulated during the development of human cancer. The best-characterized member of the family is RASSF1A, which is downregulated by promoter methylation in 40-90% of primary human tumors. We now identify and characterize a novel member of the RASSF family, RASSF6. Like the other family members, RASSF6 possesses a Ras Association domain and binds activated Ras. Exogenous expression of RASSF6 promoted apoptosis, synergized with activated K-Ras to induce cell death and inhibited the survival of specific tumor cell lines. Suppression of RASSF6 enhanced the tumorigenic phenotype of a human lung tumor cell line. Furthermore, RASSF6 is often downregulated in primary human tumors. RASSF6 shares some similar overall properties as other RASSF proteins. However, there are significant differences in biological activity between RASSF6 and other family members including a discrete tissue expression profile, cell killing specificity and impact on signaling pathways. Moreover, RASSF6 may play a role in dictating the degree of inflammatory response to the respiratory syncytial virus. Thus, RASSF6 is a novel RASSF family member that demonstrates the properties of a Ras effector and tumor suppressor but exhibits biological properties that are unique and distinct from those of other family members. PMID- 17404572 TI - Arsenic trioxide-induced neuroblastoma cell death is accompanied by proteolytic activation of nuclear Bax. AB - Arsenic trioxide (As(2)O(3)) is toxic to multidrug-resistant neuroblastoma cells in vivo and in vitro. In neuroblastoma, As(2)O(3) does not exert its cell death promoting effects via a classical apoptotic pathway. A death mechanism involving proteolytic cleavage of Bax to a p18 form seems to be of importance, because inhibition of Bax cleavage coincides with diminished cell death. As existing models of cell death implicate Bax in the intrinsic apoptotic pathway, triggering death after Bax translocation to the mitochondria, we investigated the cellular localization of p18 Bax by subcellular fractionation. After As(2)O(3) treatment, p18 Bax was only present in nuclei-enriched, mitochondria-depleted fractions. Cytoplasmic p21 Bax levels decreased, whereas total (p21 and p18) nuclear Bax increased. Overexpressed p21 Bax localized to the cytoplasm and nuclei, whereas overexpressed p18 Bax localized to extra-nuclear structures only. The inability of overexpressed p18 Bax to locate to the nucleus, and the As(2)O(3)-induced reduction of p21 Bax in the cytosol, suggest an As(2)O(3)-induced mechanism where p18 Bax gets cleaved and 'trapped' in the nucleus. This model is strengthened by the observation that calpain, the protease responsible for p18 Bax generation, is present in the nuclei, and that nuclear calpain is induced by increasing As(2)O(3) and Ca(2+) levels. PMID- 17404573 TI - Bcl-2 overexpression in PhIP-induced colon tumors: cloning of the rat Bcl-2 promoter and characterization of a pathway involving beta-catenin, c-Myc and E2F1. AB - Beta-catenin/T-cell factor (Tcf) signaling is constitutively active in the majority of human colorectal cancers, and there are accompanying changes in Bcl-2 expression. Similarly, 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine (PhIP) induced colon tumors in the rat have increased beta-catenin and elevated Bcl-2. To examine the possible direct transcriptional regulation of rat Bcl-2 by beta catenin/Tcf, we cloned and characterized the corresponding promoter region and found 70.1% similarity with its human counterpart, BCL2. Bcl-2 promoter activity was increased in response to LiCl and exogenous beta-catenin, including oncogenic mutants of beta-catenin found in PhIP-induced colon tumors. Protein/DNA arrays identified E2F1, but not beta-catenin/Tcf, as interacting most strongly with the rat Bcl-2 promoter. Exogenous E2F1 increased the promoter activity of rat Bcl-2, except in mutants lacking the E2F1 sites. As expected, beta-catenin induced its downstream target c-Myc, as well as E2F1 and Bcl-2, and this was blocked by siRNA to c-Myc or E2F1. These findings suggest an indirect pathway for Bcl-2 over expression in PhIP-induced colon tumors involving beta-catenin, c-Myc and E2F1. PMID- 17404574 TI - mir-29 regulates Mcl-1 protein expression and apoptosis. AB - Cellular expression of Mcl-1, an anti-apoptotic Bcl-2 family member, is tightly regulated. Recently, Bcl-2 expression was shown to be regulated by microRNAs, small endogenous RNA molecules that regulate protein expression through sequence specific interaction with messenger RNA. By analogy, we reasoned that Mcl-1 expression may also be regulated by microRNAs. We chose human immortalized, but non-malignant, H69 cholangiocyte and malignant KMCH cholangiocarcinoma cell lines for these studies, because Mcl-1 is dysregulated in cells with the malignant phenotype. By in silico analysis, we identified a putative target site in the Mcl 1 mRNA for the mir-29 family, and found that mir-29b was highly expressed in cholangiocytes. Interestingly, mir-29b was downregulated in malignant cells, consistent with Mcl-1 protein upregulation. Enforced mir-29b expression reduced Mcl-1 protein expression in KMCH cells. This effect was direct, as mir-29b negatively regulated the expression of an Mcl-1 3' untranslated region (UTR) based reporter construct. Enforced mir-29b expression reduced Mcl-1 cellular protein levels and sensitized the cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) cytotoxicity. Transfection of non-malignant cells (that express high levels of mir-29) with a locked-nucleic acid antagonist of mir-29b increased Mcl-1 levels and reduced TRAIL-mediated apoptosis. Thus mir 29 is an endogenous regulator of Mcl-1 protein expression, and thereby, apoptosis. PMID- 17404575 TI - Poly-(ADP-ribose) polymerase-1 (Parp-1) binds in a sequence-specific manner at the Bcl-6 locus and contributes to the regulation of Bcl-6 transcription. AB - Bcl-6 is a transcription factor that is normally expressed in germinal centre B cells. It is essential for the formation of germinal centres and the production of high-affinity antibodies. Transcriptional downregulation of Bcl-6 occurs on terminal differentiation to plasma cells. Bcl-6 is highly expressed in B-cell non Hodgkin's lymphoma and, in a subset of cases of diffuse large cell lymphoma, the mechanism of Bcl-6 overexpression involves interruption of normal transcriptional controls. Transcriptional control of Bcl-6 is, therefore, important for normal antibody responses and lymphomagenesis, but little is known of the cis-acting control elements. This report focuses on a region of mouse/human sequence homology in the first intron of Bcl-6, which is a candidate site for such a control element. We demonstrate that poly-(ADP-ribose) polymerase-1 (Parp-1) binds in vitro and in vivo to specific sequences in this region. We further show that PARP inhibitors, and Parp-1 knockdown by siRNA induce Bcl-6 mRNA expression in Bcl-6 expressing cell lines. We speculate that Parp-1 activation plays a role in switching off Bcl-6 transcription and subsequent B-cell exit from the germinal centre. PMID- 17404576 TI - Mechanisms of translational deregulation in human tumors and therapeutic intervention strategies. AB - Analysis of the recurrent genetic aberrations present in human tumors provides insight into how normal cells escape appropriate proliferation and survival cues. Commonly mutated genes encode proteins that monitor DNA damage (e.g., p53), proteins that regulate the cell cycle (such as Rb), and proteins that regulate signal transduction pathways (such as APC, PTEN and Ras). Analysis of the relevant targets and downstream events of these genes in normal and tumor cells will clearly highlight important pathways for tumorigenesis. However, more infrequent mutations are also informative in defining events critical for the process of tumorigenesis, and often delineate important pathways lying downstream of commonly mutated oncogenes and tumor suppressors. Together, these studies have led to the conclusion that deregulated protein synthesis plays an important role in human cancer. This review will discuss the evidence implicating mRNA translation as an important downstream consequence of signal transduction pathways initiated by mutated oncogenes and tumor suppressors, as well as additional genetic findings implicating the importance of global and specific translational control in human cancer. It will also discuss therapeutic strategies that take advantage of differences in translational regulation between normal and tumor cells. PMID- 17404577 TI - Id3 is a novel regulator of p27kip1 mRNA in early G1 phase and is required for cell-cycle progression. AB - P27kip is a key inhibitory protein of the cell-cycle progression, which is rapidly downregulated in early G1 phase by a post-translational mechanism involving the proteosomal degradation. In this study, using a wounding model that induces cell-cycle entry of human dermal fibroblasts, we demonstrate that p27mRNA is downregulated when cells progress into the G1 phase, and then it returns to its basal level when cells approach the S phase. By using a quantitative polymerase chain reaction screening we identified inhibitors of differentiation (Id3), a bHLH transcriptional repressor, as a candidate mediator accounting for p27 mRNA decrease. Id3 silencing, using an small interfering RNA approach, reversed the injury mediated p27 downregulation demonstrating that Id3 is involved in the transcriptional repression of p27. Reporter gene experiments and a chromatin immunoprecipitation assay showed that Id3 likely exerts its repressive action through ELK1 inhibition. By inhibiting early p27 downregulation, Id3 depletion blocked (i) the G1-phase progression as assessed by the inhibition of pRb phosphorylation and p130 degradation and (ii) the G1/S transition as observed by the inhibition of cyclin A induction, demonstrating that p27 mRNA decrease is required for cell proliferation. Apart from its effect on the early p27 diminution, Id3 appears also involved in the control of the steady-state level of p27 at the G1/S boundary. In conclusion, this study identifies a novel mechanism of p27 regulation which besides p27 protein degradation also implicates a transcriptional mechanism mediated by Id3. PMID- 17404578 TI - Expression of activation-induced cytidine deaminase in human hepatocytes via NF kappaB signaling. AB - Activation-induced cytidine deaminase (AID) is involved in somatic DNA alterations of the immunoglobulin gene for amplification of immune diversity. The fact that constitutive expression of AID in mice causes tumors in various organs, including lymphoid tissues and lungs, suggests the important role of the aberrant editing activity of AID on various tumor-related genes for carcinogenesis. AID expression, however, is restricted to activated B cells under physiological conditions. We demonstrate here that ectopic AID expression is induced in response to tumor necrosis factor-alpha stimulation in cultured human hepatocytes. The proinflammatory cytokine-mediated expression of AID is achieved by IkappaB kinase-dependent nuclear factor (NF)-kappaB signaling pathways. Hepatitis C virus, one of the leading causes of hepatocellular carcinoma (HCC), enhanced AID expression via NF-kappaB activation through expression of viral core protein. The aberrant expression of AID in hepatoma-derived cells resulted in accumulation of genetic alterations in the c-myc and pim1 genes, suggesting that inappropriate expression of AID acts as a DNA mutator that enhances the genetic susceptibility to mutagenesis in human hepatocytes. Our current findings indicate that the inappropriate expression of AID is induced by proinflammatory cytokine stimulation and may provide the link between hepatic inflammation and the development of HCC. PMID- 17404579 TI - Variability in human hepatic MRP4 expression: influence of cholestasis and genotype. AB - The multidrug resistance protein 4 (MRP4) is an efflux transporter involved in the transport of endogenous substrates and xenobiotics. We measured MRP4 mRNA and protein expression in human livers and found a 38- and 45-fold variability, respectively. We sequenced 2 kb of the 5'-flanking region, all exons and intron/exon boundaries of the MRP4 gene in 95 patients and identified 74 genetic variants including 10 non-synonymous variations, seven of them being located in highly conserved regions. None of the detected polymorphisms was significantly associated with changes in the MRP4 mRNA or protein expression. Immunofluorescence microscopy indicated that none of the non-synonymous variations affected the cellular localization of MRP4. However, in cholestatic patients the MRP4 mRNA and protein expression both were significantly upregulated compared to non-cholestatic livers (protein: 299+/-138 vs 100+/-60a.u., P<0.001). Taken together, human hepatic MRP4 expression is highly variable. Genetic variations were not sufficient to explain this variability. In contrast, cholestasis is one major determinant of human hepatic MRP4 expression. PMID- 17404580 TI - Polymorphisms of norepinephrine transporter and adrenergic receptor alpha1D are associated with the response to beta-blockers in dilated cardiomyopathy. AB - Recent clinical trials have clearly demonstrated that the administration with beta-blockers decreases the mortality in the patients with chronic heart failure (CHF). However, significant heterogeneity exists in the effectiveness of beta blockers among individual cases. We focused on 39 polymorphisms in 16 genes related to adrenergic system and investigated their association with the response to beta-blockers among 80 patients with CHF owing to idiopathic dilated cardiomyopathy. The polymorphisms of NET T-182C (P=0.019), ADRA1D T1848A (P=0.023) and ADRA1D A1905G (P=0.029) were associated with the improvement of left ventricular fractional shortening (LVFS) by beta-blockers. Furthermore, combined genotype analysis of NET T-182C and ADRA1D T1848A revealed a significant difference in LVFS improvement among genotype groups (P=0.011). These results suggest that NET (T-182C) and ADRA1D (T1848A and A1905G) polymorphisms are predictive markers of the response to beta-blockers. Genotyping of these polymorphisms may provide clinical insights into an individual difference in the response to the beta-blocker therapy in CHF. PMID- 17404581 TI - A comparative study of serum protein-bound sialic acid in benign and malignant prostatic growth: possible role of oxidative stress in sialic acid homeostasis. AB - Benign and malignant prostatic growths are associated with an increase in sialoconjugates (e.g. prostate-specific antigen (PSA)) in blood. Oxidative stress plays a crucial role in pathogenesis of various malignancies. The objective of this study was to evaluate oxidative stress parameters and protein-bound sialic acid level in sera of prostatic tumor cases and to asses for any association between them. Sera samples were collected and estimated for carbonylation of proteins, lipid peroxidation products, PSA and protein-bound sialic acid from 10 patients in each group with prostatic carcinoma (Ca prostate) and benign prostatic hyperplasia (BPH) along with 10 healthy male subjects of similar age group as control. In carcinoma prostate cases, lipid peroxides, protein carbonyls, protein-bound sialic acid and PSA were significantly increased compared to BPH and controls. There was significant association between oxidative stress parameters (lipid peroxide and protein carbonyl) and sialoconjugates (PSA and protein-bound sialic acid). In BPH cases, serum lipid peroxides and protein bound sialic acid were significantly higher in comparison to controls and protein carbonyls were correlated with protein-bound sialic acid. ROC curve for sialic acid showed that it can be used as a marker to differentiate carcinoma prostate from benign growth of prostate at a cutoff level of 11.38 mug/mg protein with a sensitivity of 100% and specificity of 80%. We conclude that oxidative stress might be associated with the degree of sialylation of protein and graded changes in these parameters possibly unveil the pathogenic demarcation from benign to malignant condition of prostate. PMID- 17404582 TI - Docetaxel-induced Meibomian duct inflammation and blockage leading to chalazion formation. AB - We report a 71-year male with castration-resistant metastatic prostate cancer who was treated with weekly docetaxel for 12 weeks and developed significant eye irritation and dryness during treatment. Subsequently, the patient presented with a lower eyelid mass, which on excision was demonstrated to be a chalazion. Docetaxel induced Meibomian duct inflammation and blockage is the likely cause of this presentation in a patient with no history of eyelid masses in the past. PMID- 17404584 TI - Hybrid necrosis: autoimmunity as a potential gene-flow barrier in plant species. AB - Ecological factors, hybrid sterility and differences in ploidy levels are well known for contributing to gene-flow barriers in plants. Another common postzygotic incompatibility, hybrid necrosis, has received comparatively little attention in the evolutionary genetics literature. Hybrid necrosis is associated with a suite of phenotypic characteristics that are similar to those elicited in response to various environmental stresses, including pathogen attack. The genetic architecture is generally simple, and complies with the Bateson Dobzhansky-Muller model for hybrid incompatibility between species. We survey the extensive literature on this topic and present the hypothesis that hybrid necrosis can result from autoimmunity, perhaps as a pleiotropic effect of evolution of genes that are involved in pathogen response. PMID- 17404585 TI - Back to basics: food for thought--and nutrition. PMID- 17404586 TI - Is the prevalence of idiopathic ulcers really on the increase? PMID- 17404587 TI - Laparoscopic gastric banding or gastric bypass for morbid obesity? PMID- 17404588 TI - Current therapy for autoimmune hepatitis. AB - The treatment of autoimmune hepatitis is evolving as the pathogenic pathways that underlie the disease are defined, new immunosuppressive agents are tested, and site-specific molecular interventions become feasible. Prednisone alone or at a reduced dose combined with azathioprine is the conventional treatment. Patients with HLA genotype DRB1*0301 have a poorer treatment response and a more frequent need for liver transplantation than those with HLA genotype DRB1*0401. Therapy to the point when liver test results and histological findings are normal reduces, but does not eliminate, the occurrence of relapse. Treatment failure warrants reassessment with regard to the accuracy of the original diagnosis and the exclusion of variant forms of hepatitis or concomitant alternative diseases. Ciclosporin might be effective as short-term, front-line therapy in infants and adults, and calcineurin inhibitors might salvage patients who are refractory to corticosteroid regimens. Mycophenolate mofetil can induce an improvement in laboratory test results and reduce the requirement for corticosteroids. Sirolimus is effective for treatment of de novo autoimmune hepatitis that develops after liver transplantation. Synthetic peptides that block autoantigen presentation, cytokine manipulations, oral tolerance regimens, T-cell vaccination, and gene therapy are all interventions that will be able to emerge after a reliable animal model of the human disease has been developed. PMID- 17404589 TI - Technology insight: advances in liver imaging. AB - The role of diagnostic imaging in the assessment of liver disease continues to gain in importance. The classic techniques used for liver imaging are ultrasonography, CT and MRI. In the past decade, there have been significant advances in all three techniques. In this article, we discuss the advances in ultrasonography, CT and MRI that have improved assessment of focal and diffuse liver disease, including the development of hardware, software, processing algorithms and procedural innovations. PMID- 17404590 TI - Colonic infiltration with chronic myelomonocytic leukemia. AB - BACKGROUND: A 75-year-old female presented with a 1 month history of non-bloody diarrhea, associated with abdominal cramping and urgency. Her medical history was notable for chronic myelomonocytic leukemia, diagnosed 6 years previously and managed expectantly by monitoring the patient's complete blood count. Over several months, the patient's symptoms progressed, which resulted in significant weight loss. The patient's course of disease was ultimately complicated by acute disseminated encephalomyelitis and death. INVESTIGATIONS: Physical examination, laboratory investigations, stool studies, colonoscopy with biopsies, immunohistochemistry and pathologic review of biopsy specimens. DIAGNOSIS: Leukemic colitis. MANAGEMENT: Management of underlying leukemia with systemic hydroxyurea and topical colonic 5-aminosalicylic acid therapy. PMID- 17404591 TI - Models for antigen receptor gene rearrangement: CDR3 length. AB - Despite the various processing steps involved in V(D)J recombination, which could potentially introduce many biases in the length distribution of complementarity determining region 3 (CDR3) segments, the observed CDR3 length distributions for complete repertoires are very close to a normal-like distribution. This raises the question of whether this distribution is simply a result of the random steps included in the process of gene rearrangement, or has been optimized during evolution. We have addressed this issue by constructing a simulation of gene rearrangement, which takes into account the DNA modification steps included in the process, namely hairpin opening, nucleotide additions, and nucleotide deletions. We found that the near-Gaussian- shape of CDR3 length distribution can only be obtained under a relatively narrow set of parameter values, and thus our model suggests that specific biases govern the rearrangement process. In both B cell receptor (BCR) heavy chain and T-cell receptor beta chain, we obtained a Gaussian distribution using identical parameters, despite the difference in the number and the lengths of the D segments. Hence our results suggest that these parameters most likely reflect the optimal conditions under which the rearrangement process occurs. We have subsequently used the insights gained in this study to estimate the probability of occurrence of two exactly identical BCRs over the course of a human lifetime. Whereas identical rearrangements of the heavy chain are highly unlikely to occur within one human lifetime, for the light chain we found that this probability is not negligible, and hence the light chain CDR3 alone cannot serve as an indicator of B-cell clonality. PMID- 17404592 TI - Anti-BDCA-4 (neuropilin-1) antibody can suppress virus-induced IFN-alpha production of plasmacytoid dendritic cells. AB - Plasmacytoid dendritic cells (PDC) in human blood are the main source of virus induced interferon (IFN)-alpha. They exhibit a lineage-negative phenotype but all express BDCA-4, which is homologous to the neuronal receptor neuropilin-1. Specific staining with anti-BDCA-4 antibody is used for positive isolation of PDC from blood by magnetic cells sorting. Here, it is demonstrated that these positively selected PDC showed reduced or completely abolished IFN-alpha release compared to unstained PDC, which were negatively selected by magnetic depletion of lineage-positive blood mononuclear cells. In addition, treatment of these unstained PDC with anti-BDCA-4 mAb also resulted in at least two-fold lower or reduced virus-induced IFN-alpha production. It is shown that the antibody not only affects cell survival or block virus attachment but also reduces IFN-alpha release induced by non-viral CpG oligodeoxynucleotides. In conclusion, data suggest an immunoregulatory role for BDCA-4 on PDC as demonstrated for IFN-alpha response to virus. PMID- 17404593 TI - Xom interacts with and stimulates transcriptional activity of LEF1/TCFs: implications for ventral cell fate determination during vertebrate embryogenesis. AB - LEF1/TCFs are high mobility group box-containing transcriptional factors mediating canonical Wnt/beta-catenin signaling during early embryogenesis and tumorigenesis. Beta-catenin forms a complex with LEF1/TCFs and transactivates LEF1/TCF-mediated transcriptions during dorsalization. Although LEF-mediated transcription is also implicated in ventralization, the underlying molecular mechanism is not well understood. Using the vertebrate Xenopus laevis model system, we found that Xom, which is a ventralizing homeobox protein with dual roles of transcriptional activation and repression, forms a complex with LEF1/TCF through its homeodomain and transactivates LEF1/TCF-mediated transcription through its N-terminal transactivation domain (TAD). Our data show that Xom lacking the N-terminal TAD fails to transactivate ventral genes, such as BMP4 and Xom itself, but retains the ability to suppress transcriptional activation of dorsal gene promoters, such as the Goosecoid promoter, indicating that transactivation and repression are separable functions of Xom. It has been postulated that Xom forms a positive re-enforcement loop with BMP4 to promote ventralization and to suppress dorsal gene expression. Consistent with an essential role of Xom transactivation of LEF1/TCFs during early embryogenesis, we found that expression of the dominant-negative Xom mutant that lacks the TAD fails to re-enforce the ventral signaling of BMP4 and causes a catastrophic effect during gastrulation. Our data suggest that the functional interaction of Xom and LEF1/TCF-factors is essential for ventral cell fate determination and that LEF1/TCF factors may function as a point of convergence to mediate the combined signaling of Wnt/beta-catenin and BMP4/Xom pathways during early embryogenesis. PMID- 17404594 TI - p53 and Bad: remote strangers become close friends. PMID- 17404595 TI - Shp2, a novel oncogenic tyrosine phosphatase and potential therapeutic target for human leukemia. PMID- 17404596 TI - CTTN (EMS1): an oncogene contributing to the metastasis of esophageal squamous cell carcinoma. PMID- 17404597 TI - Axin bridges Daxx to p53. PMID- 17404598 TI - Use of arrays to investigate the contribution of ATP-binding cassette transporters to drug resistance in cancer chemotherapy and prediction of chemosensitivity. AB - Multidrug resistance (MDR) is a major problem in cancer chemotherapy. One of the best known mechanisms of MDR is the elevated expression of ATP-binding cassette (ABC) transporters. While some members of human ABC transporters have been shown to cause drug resistance with elevated expression, it is not yet known whether the over-expression of other members could also contribute to drug resistance in many model cancer cell lines and clinics. The recent development of microarrays and quantitative PCR arrays for expression profiling analysis of ABC transporters has helped address these issues. In this article, various arrays with limited or full list of ABC transporter genes and their use in identifying ABC transporter genes in drug resistance and chemo-sensitivity prediction will be reviewed. PMID- 17404599 TI - Adiponectin as a negative regulator in obesity-related mammary carcinogenesis. PMID- 17404600 TI - Antibody gene therapy: an attractive approach for the treatment of cancers and other chronic diseases. PMID- 17404601 TI - Up-regulation of Bcl-2 is required for the progression of prostate cancer cells from an androgen-dependent to an androgen-independent growth stage. AB - Bcl-2 is an anti-apoptotic oncoprotein and its protein levels are inversely correlated with prognosis in many cancers. However, the role of Bcl-2 in the progression of prostate cancer is not clear. Here we report that Bcl-2 is required for the progression of LNCaP prostate cancer cells from an androgen dependent to an androgen-independent growth stage. The mRNA and protein levels of Bcl-2 are significantly increased in androgen-independent prostate cancer cells. shRNA-mediated gene silencing of Bcl-2 in androgen-independent prostate cancer cells promotes UV-induced apoptosis and suppresses the growth of prostate tumors in vivo. Growing androgen-dependent cells under androgen-deprivation conditions results in formation of androgen-independent colonies; and the transition from androgen-dependent to androgen-independent growth is blocked by ectopic expression of the Bcl-2 antagonist Bax or Bcl-2 shRNA. Thus, our results demonstrate that Bcl-2 is not only critical for the survival of androgen independent prostate cancer cells, but is also required for the progression of prostate cancer cells from an androgen-dependent to an androgen-independent growth stage. PMID- 17404602 TI - Synthetic Studies Toward Bryostatin 1: Preparation of a C(1)-C(16) Fragment by Pyran Annulation. AB - An expeditious assembly of a C(1)-C(16) subunit of bryostatin 1 is described. A pyran annulation reaction was utilized to form the B-ring by reaction of a hydroxy-allylsilane with a fully elaborated A-ring subunit. This annulation process proceeded with complete diastereoselectivity and in excellent isolated yield despite the presence of potentially sensitive functionality in the A-ring segment. PMID- 17404603 TI - Adolescent Ego-Development Trajectories and Young Adult Relationship Outcomes. AB - Adolescent ego-development trajectories were related to close-relationship outcomes in young adulthood. An adolescent sample completed annual measures of ego development from ages 14 through 17. The authors theoretically determined and empirically traced five ego-development trajectories reflecting stability or change. At age 25, the sample completed a close-relationship interview and consented for two peers to rate the participants'ego resiliency and hostility. Participants who followed the profound-arrest trajectory in adolescence reported more mundane sharing of experiences, more impulsive or egocentric conflict resolution tactics, and less mature interpersonal understanding in their young adult relationships, and their young adult peers described these participants as more hostile. Participants who attained or maintained higher levels of ego development in adolescence reported more complex sharing of experiences, more collaborative conflict-resolution strategies, and greater interpersonal understanding, and their young adult peers rated them as less hostile and as more flexible. PMID- 17404604 TI - Does Non-Exercise Activity Thermogenesis Contribute to Non-Shivering Thermogenesis? AB - We wanted to examine if spontaneous physical activity contributes to non shivering thermogenesis. Ten lean, healthy male subjects wore a physical activity, micro-measurement system whilst the room temperature was randomly altered at two hourly intervals between thermoneutral (72 degrees F), cool (62 degrees F) and warm (82 degrees F) temperatures. Physical activity measured during the thermoneutral, cooling and warming periods was not significantly different. Cooling, increased EE above basal and thermoneutral values 2061 +/- 344 kcal/day (p <0.01). Thus, the increase in energy expenditure associated with short-term environmental cooling in lean, healthy males does not appear to be due to increased spontaneous physical activity or fidgeting. PMID- 17404605 TI - Continuous-wave Submillimeter-wave Gyrotrons. AB - Recently, dynamic nuclear polarization enhanced nuclear magnetic resonance (DNP/NMR) has emerged as a powerful technique to obtain significant enhancements in spin spectra from biological samples. For DNP in modern NMR systems, a high power continuous-wave source in the submillimeter wavelength range is necessary. Gyrotrons can deliver tens of watts of CW power at submillimeter wavelengths and are well suited for use in DNP/NMR spectrometers. To date, 140 GHz and 250 GHz gyrotrons are being employed in DNP spectrometer experiments at 200 MHz and 380 MHz at MIT. A 460 GHz gyrotron, which has operated with 8 W of CW output power, will soon be installed in a 700 MHz NMR spectrometer. High power radiation with good spectral and spatial resolution from these gyrotrons should provide NMR spectrometers with high signal enhancement through DNP. Also, these tubes operating at submillimeter wavelengths should have important applications in research in physics, chemistry, biology, materials science and medicine. PMID- 17404606 TI - A Model of 3D Propagation in Discrete Cardiac Tissue. AB - A model was developed of a bundle of cardiac fibers embedded in an extracellular space. In contrast to the classical bidomain approach, the model is constructed such that the intracellular and extracellular spaces are spatially distinct. The model was used to test the hypothesis that the distribution of the extracellular fluid in the tissue can affect the conduction velocity. The preliminary results suggest that under nominally normal conditions, the propagation speed depends on the fraction of extracellular space and less on the actual distribution of extracellular space within the bundle. PMID- 17404607 TI - aCGHViewer: a generic visualization tool for aCGH data. AB - Array-Comparative Genomic Hybridization (aCGH) is a powerful high throughput technology for detecting chromosomal copy number aberrations (CNAs) in cancer, aiming at identifying related critical genes from the affected genomic regions. However, advancing from a dataset with thousands of tabular lines to a few candidate genes can be an onerous and time-consuming process. To expedite the aCGH data analysis process, we have developed a user-friendly aCGH data viewer (aCGHViewer) as a conduit between the aCGH data tables and a genome browser. The data from a given aCGH analysis are displayed in a genomic view comprised of individual chromosome panels which can be rapidly scanned for interesting features. A chromosome panel containing a feature of interest can be selected to launch a detail window for that single chromosome. Selecting a data point of interest in the detail window launches a query to the UCSC or NCBI genome browser to allow the user to explore the gene content in the chromosomal region. Additionally, aCGHViewer can display aCGH and expression array data concurrently to visually correlate the two. aCGHViewer is a stand alone Java visualization application that should be used in conjunction with separate statistical programs. It operates on all major computer platforms and is freely available at http://falcon.roswellpark.org/aCGHview/. PMID- 17404608 TI - Genetic analysis of human hematological malignancies rewarded. PMID- 17404609 TI - gp130 receptor ligands as potential therapeutic targets for obesity. AB - Obesity and its related cluster of pathophysiologic conditions including insulin resistance, glucose intolerance, dyslipidemia, and hypertension are recognized as growing threats to world health. It is now estimated that 10% of the world's population is overweight or obese. As a result, new therapeutic options for the treatment of obesity are clearly warranted. Recent research has focused on the role that gp130 receptor ligands may play as potential therapeutic targets in obesity. One cytokine in particular, ciliary neurotrophic factor (CNTF), acts both centrally and peripherally and mimics the biologic actions of the appetite control hormone leptin, but unlike leptin, CNTF appears to be effective in obesity and as such may have therapeutic potential. In addition, CNTF suppresses inflammatory signaling cascades associated with lipid accumulation in liver and skeletal muscle. This review examines the potential role of gp130 receptor ligands as part of a therapeutic strategy to treat obesity. PMID- 17404610 TI - Sickle cell disease: old discoveries, new concepts, and future promise. AB - The discovery of the molecular basis of sickle cell disease was an important landmark in molecular medicine. The modern tools of molecular and cellular biology have refined our understanding of its pathophysiology and facilitated the development of new therapies. In this review, we discuss some of the important advances in this field and the impediments that limit the impact of these advances. PMID- 17404611 TI - Putting pancreatic cell plasticity to the test. AB - Diabetes results from the absolute or relative deficiency of insulin-producing beta cells. The prospect that non-beta pancreatic cells could be harnessed to become beta cells has led to interest in understanding the plasticity of pancreatic cells. Recent studies, however, have shown that adult beta cells are largely derived from preexisting beta cells. In this issue of the JCI, Desai et al. show that acinar cells, the major cell type in the pancreas, do not contribute to new beta cells formed during pancreatic regeneration (see the related article beginning on page 971). These studies suggest that the fate of adult pancreatic cell lineages is immutable. However, also in this issue of the JCI, Collombat et al. demonstrate that inducing a single transcription factor named Arx in adult beta cells causes these cells to undergo massive transdifferentiation into alpha and pancreatic polypeptide endocrine cells (see the related article beginning on page 961). This finding points to an unexpected plasticity of postnatal pancreatic endocrine cells. PMID- 17404612 TI - HIF-1 and HIF-2: working alone or together in hypoxia? AB - Erythropoietin (EPO) is the hormonal regulator of red cell production and provided the paradigm for oxygen-regulated gene expression that led to the discovery of hypoxia-inducible factor (HIF). In this issue of the JCI, Rankin and colleagues show, using targeted gene inactivation, that induction of Epo expression in murine liver is dependent on the integrity of HIF-2alpha, and not HIF-1alpha (see the related article beginning on page 1068). These results demonstrate distinct functions for different HIF-alpha isoforms that could potentially be exploited in therapeutic approaches to anemia. PMID- 17404613 TI - HOX deregulation in acute myeloid leukemia. AB - The deregulation of homeobox (HOX) genes in acute myeloid leukemia (AML) and the potential for these master regulators to perturb normal hematopoiesis is well established. To date, overexpression of HOX genes in AML has been attributed to specific chromosomal aberrations and abnormalities involving mixed-lineage leukemia (MLL), an upstream regulator of HOX genes. The finding reported in this issue of the JCI by Scholl et al. that caudal-type homeobox transcription factor 2 (CDX2), which is capable of affecting HOX gene expression during embryogenesis, is overexpressed in 90% of patients with AML and induces a transplantable AML in murine models provides an alternative mechanism for HOX-induced leukemogenesis and yields important insights into the hierarchy of HOX gene regulation in AML (see the related article beginning on page 1037). PMID- 17404614 TI - Hypoglycemia, functional brain failure, and brain death. AB - Hypoglycemia commonly causes brain fuel deprivation, resulting in functional brain failure, which can be corrected by raising plasma glucose concentrations. Rarely, profound hypoglycemia causes brain death that is not the result of fuel deprivation per se. In this issue of the JCI, Suh and colleagues use cell culture and in vivo rodent studies of glucose deprivation and marked hypoglycemia and provide evidence that hypoglycemic brain neuronal death is in fact increased by neuronal NADPH oxidase activation during glucose reperfusion (see the related article beginning on page 910). This finding suggests that, at least in the setting of profound hypoglycemia, therapeutic hyperglycemia should be avoided. PMID- 17404615 TI - Yin and yang interplay of IFN-gamma in inflammation and autoimmune disease. AB - IFN-gamma has long been recognized as a signature proinflammatory cytokine that plays a central role in inflammation and autoimmune disease. There is now emerging evidence indicating that IFN-gamma possesses unexpected properties as a master regulator of immune responses and inflammation. In this issue of the JCI, Guillonneau et al. show that indefinite allograft survival induced by CD40Ig treatment is mediated by CD8(+)CD45RC(low) T cells through the production of IFN gamma (see the related article beginning on page 1096), supporting the emerging view that IFN-gamma is critical in the self-regulation of inflammation. These contradictory roles of IFN-gamma, perhaps best understood by the principle of yin and yang, represent one of nature's paradoxes, whereby the same cytokine functions as an inducer as well as a regulator for inflammation. Understanding this complex process of IFN-gamma signaling is essential, as it has therapeutic implications. PMID- 17404616 TI - The renin-angiotensin system: it's all in your head. AB - Components of the renin-angiotensin system (RAS) are expressed in a number of areas in the brain involved in cardiovascular control. However, it has been difficult to link RAS actions in circumscribed brain regions to specific physiological functions. In a study appearing in this issue of the JCI, Sakai and associates use a combination of sophisticated transgenic techniques and stereotaxic microinjection of recombinant viral vectors to demonstrate a pivotal role in the regulation of thirst and salt appetite of angiotensin II generated in the subfornical organ in the brain (see the related article beginning on page 1088). PMID- 17404617 TI - Hypoglycemic neuronal death is triggered by glucose reperfusion and activation of neuronal NADPH oxidase. AB - Hypoglycemic coma and brain injury are potential complications of insulin therapy. Certain neurons in the hippocampus and cerebral cortex are uniquely vulnerable to hypoglycemic cell death, and oxidative stress is a key event in this cell death process. Here we show that hypoglycemia-induced oxidative stress and neuronal death are attributable primarily to the activation of neuronal NADPH oxidase during glucose reperfusion. Superoxide production and neuronal death were blocked by the NADPH oxidase inhibitor apocynin in both cell culture and in vivo models of insulin-induced hypoglycemia. Superoxide production and neuronal death were also blocked in studies using mice or cultured neurons deficient in the p47(phox) subunit of NADPH oxidase. Chelation of zinc with calcium disodium EDTA blocked both the assembly of the neuronal NADPH oxidase complex and superoxide production. Inhibition of the hexose monophosphate shunt, which utilizes glucose to regenerate NADPH, also prevented superoxide formation and neuronal death, suggesting a mechanism linking glucose reperfusion to superoxide formation. Moreover, the degree of superoxide production and neuronal death increased with increasing glucose concentrations during the reperfusion period. These results suggest that high blood glucose concentrations following hypoglycemic coma can initiate neuronal death by a mechanism involving extracellular zinc release and activation of neuronal NADPH oxidase. PMID- 17404618 TI - Involvement of PLEKHM1 in osteoclastic vesicular transport and osteopetrosis in incisors absent rats and humans. AB - This study illustrates that Plekhm1 is an essential protein for bone resorption, as loss-of-function mutations were found to underlie the osteopetrotic phenotype of the incisors absent rat as well as an intermediate type of human osteopetrosis. Electron and confocal microscopic analysis demonstrated that monocytes from a patient homozygous for the mutation differentiated into osteoclasts normally, but when cultured on dentine discs, the osteoclasts failed to form ruffled borders and showed little evidence of bone resorption. The presence of both RUN and pleckstrin homology domains suggests that Plekhm1 may be linked to small GTPase signaling. We found that Plekhm1 colocalized with Rab7 to late endosomal/lysosomal vesicles in HEK293 and osteoclast-like cells, an effect that was dependent on the prenylation of Rab7. In conclusion, we believe PLEKHM1 to be a novel gene implicated in the development of osteopetrosis, with a putative critical function in vesicular transport in the osteoclast. PMID- 17404619 TI - Embryonic endocrine pancreas and mature beta cells acquire alpha and PP cell phenotypes upon Arx misexpression. AB - Aristaless-related homeobox (Arx) was recently demonstrated to be involved in pancreatic alpha cell fate specification while simultaneously repressing the beta and delta cell lineages. To establish whether Arx is not only necessary, but also sufficient to instruct the alpha cell fate in endocrine progenitors, we used a gain-of-function approach to generate mice conditionally misexpressing this factor. Mice with forced Arx expression in the embryonic pancreas or in developing islet cells developed a dramatic hyperglycemia and eventually died. Further analysis demonstrated a drastic loss of beta and delta cells. Concurrently, a remarkable increase in the number of cells displaying alpha cell or, strikingly, pancreatic polypeptide (PP) cell features was observed. Notably, the ectopic expression of Arx induced in embryonic or adult beta cells led to a loss of the beta cell phenotype and a concomitant increase in a number of cells with alpha or PP cell characteristics. Combining quantitative real-time PCR and lineage-tracing experiments, we demonstrate that, in adult mice, the misexpression of Arx, rather than its overexpression, promotes a conversion of beta cells into glucagon- or PP-producing cells in vivo. These results provide important insights into the complex mechanisms underlying proper pancreatic endocrine cell allocation and cell identity acquisition. PMID- 17404620 TI - Preexisting pancreatic acinar cells contribute to acinar cell, but not islet beta cell, regeneration. AB - It has been suggested that pancreatic acinar cells can serve as progenitors for pancreatic islets, a concept with substantial implications for therapeutic efforts to increase insulin-producing beta cell mass in patients with diabetes. We report what we believe to be the first in vivo lineage tracing approach to determine the plasticity potential of pancreatic acinar cells. We developed an acinar cell-specific inducible Cre recombinase transgenic mouse, which, when mated with a reporter strain and pulsed with tamoxifen, resulted in permanent and specific labeling of acinar cells and their progeny. During various time periods of observation and using several models to provoke injury, we failed to observe any chase of the labeled cells into the endocrine compartment, indicating that acinar cells do not normally transdifferentiate into islet beta cells in vivo in adult mice. In contrast, we observed a substantial role for replication of preexisting acinar cells in the regeneration of new acinar cells after partial pancreatectomy. These results indicate that mature acinar cells harbor a facultative acinar but not endocrine progenitor capacity. PMID- 17404621 TI - Hypoxia-inducible factor-2 (HIF-2) regulates hepatic erythropoietin in vivo. AB - Erythropoiesis is critically dependent on erythropoietin (EPO), a glycoprotein hormone that is regulated by hypoxia-inducible factor (HIF). Hepatocytes are the primary source of extrarenal EPO in the adult and express HIF-1 and HIF-2, whose roles in the hypoxic induction of EPO remain controversial. In order to define the role of HIF-1 and HIF-2 in the regulation of hepatic EPO expression, we have generated mice with conditional inactivation of Hif-1alpha and/or Hif-2alpha (Epas1) in hepatocytes. We have previously shown that inactivation of the von Hippel-Lindau tumor suppressor pVHL, which targets both HIFs for proteasomal degradation, results in increased hepatic Epo production and polycythemia independent of Hif-1alpha. Here we show that conditional inactivation of Hif 2alpha in pVHL-deficient mice suppressed hepatic Epo and the development of polycythemia. Furthermore, we found that physiological Epo expression in infant livers required Hif-2alpha but not Hif-1alpha and that the hypoxic induction of liver Epo in anemic adults was Hif-2alpha dependent. Since other Hif target genes such phosphoglycerate kinase 1 (Pgk) were Hif-1alpha dependent, we provide genetic evidence that HIF-1 and HIF-2 have distinct roles in the regulation of hypoxia-inducible genes and that EPO is preferentially regulated by HIF-2 in the liver. PMID- 17404622 TI - Local production of angiotensin II in the subfornical organ causes elevated drinking. AB - The mechanism controlling cell-specific Ang II production in the brain remains unclear despite evidence supporting neuron-specific renin and glial- and neuronal specific angiotensinogen (AGT) expression. We generated double-transgenic mice expressing human renin (hREN) from a neuron-specific promoter and human AGT (hAGT) from its own promoter (SRA mice) to emulate this expression. SRA mice exhibited an increase in water and salt intake and urinary volume, which were significantly reduced after chronic intracerebroventricular delivery of losartan. Ang II-like immunoreactivity was markedly increased in the subfornical organ (SFO). To further evaluate the physiological importance of de novo Ang II production specifically in the SFO, we utilized a transgenic mouse model expressing a floxed version of hAGT (hAGT(flox)), so that deletions could be induced with Cre recombinase. We targeted SFO-specific ablation of hAGT(flox) by microinjection of an adenovirus encoding Cre recombinase (AdCre). SRA(flox) mice exhibited a marked increase in drinking at baseline and a significant decrease in water intake after administration of AdCre/adenovirus encoding enhanced GFP (AdCre/AdEGFP), but not after administration of AdEGFP alone. This decrease only occurred when Cre recombinase correctly targeted the SFO and correlated with a loss of hAGT and angiotensin peptide immunostaining in the SFO. These data provide strong genetic evidence implicating de novo synthesis of Ang II in the SFO as an integral player in fluid homeostasis. PMID- 17404623 TI - CD40Ig treatment results in allograft acceptance mediated by CD8CD45RC T cells, IFN-gamma, and indoleamine 2,3-dioxygenase. AB - Treatment with CD40Ig results in indefinite allograft survival in a complete MHC mismatched heart allograft model in the rat. Here we show that serial second, third, and fourth adoptive transfers of total splenocytes from CD40Ig-treated recipients into secondary recipients led to indefinite donor-specific allograft acceptance. Purification of splenocyte subpopulations from CD40Ig-treated recipients demonstrated that only the adoptively transferred CD8(+)CD45RC(low) subset resulted in donor-specific long-term survival, whereas CD8(+)CD45RC(low) T cells from naive animals did not. Accepted grafts displayed increased indoleamine 2,3-dioxygenase (IDO) expression restricted in the graft to ECs. Coculture of donor ECs with CD8(+)CD45RC(low) T cells purified from CD40Ig-treated animals resulted in donor-specific IDO expression dependent on IFN-gamma. Neutralization of IFN-gamma or IDO triggered acute allograft rejection in both CD40Ig-treated and adoptively transferred recipients. This study demonstrates for what we believe to be the first time that interference in CD40-CD40 ligand (CD40-CD40L) interactions induces allospecific CD8(+) Tregs that maintain allograft survival. CD8(+)CD45RC(low) T cells act through IFN-gamma production, which in turn induces IDO expression by graft ECs. Thus, donor alloantigen-specific CD8(+) Tregs may promote local graft immune privilege through IDO expression. PMID- 17404624 TI - 2006 Association of American Physicians George M. Kober Medal. Introduction of David G. Nathan, MD. PMID- 17404625 TI - Acceptance of the 2006 Kober medal. PMID- 17404626 TI - Introduction to the special issue in honour of Vincenzo Balzani "Smart photoactive molecules, supermolecules, materials". PMID- 17404627 TI - Using light to induce energy and electron transfer or molecular motions in multicomponent systems. AB - Light-induced processes are at the basis of fundamental natural phenomena as well as of a variety of applications. Since the functions that can arise from the interaction between light and matter depend on the degree of complexity and organization of the receiving 'matter', the research on these processes has progressively moved from molecular to supramolecular (multicomponent) systems, thereby originating the field of supramolecular photochemistry. In this context, examples of photochemical molecular devices and machines--that is, multicomponent chemical systems capable to perform specific functions under light stimulation- have been developed. Here we report examples of molecular devices in solution, where light is employed (i) to create and transport electronic energy, (ii) to displace electrons, and (iii) to set molecular components in motion, mimicking the operation of mechanical machines and motors. These studies are of interest not only for increasing the basic understanding and testing of current theoretical treatments of photoinduced processes but also eventually for the growth of nanoscience. PMID- 17404628 TI - Luminescence of a Pt(II) complex in the presence of DNA. Dependence of luminescence changes on the interaction binding mode. AB - Luminescence intensity changes of a Pt(II) complex which is known to bind externally to DNA at low [DNA]/[complex] ratio and to intercalate at high [DNA]/[complex] ratio are studied in the presence of calf thymus DNA. External binding is demonstrated to induce luminescence enhancement whereas intercalation leads to luminescence quenching. The reasons for this behaviour are discussed. PMID- 17404629 TI - Photochemical tuning of light emission in a conjugated polymer containing norbornadiene units in the main chain. AB - A conjugated alternating copolymer containing norbornadiene and bis(ethynylene)phenylene units was prepared by the Cassar-Heck-Sonogashira cross coupling reaction. Its electroluminescence was tested in a device, and its fluorescence colour could be tuned by light-induced norbornadiene-quadricyclane isomerization. PMID- 17404630 TI - Synthesis, structure and luminescence studies of heterometallic gold(I)-copper(I) and -silver(I) alkynyl clusters/aggregates. AB - A series of pentanuclear gold(I)-copper(I) and -silver(I) mixed-metal alkynyl complexes, [(n)Bu(4)N][Au(3)M(2)(C triple bond CC(6)H(4)R-p)(6)] [M = Cu, R = OMe, O(n)Bu, O(n)Hex, Me, Et; M = Ag, R = Et, O(n)Hex] have been synthesized. The complexes were found to be emissive both in the solid state and in fluid solutions. DFT calculations at the B3LYP level of theory were performed on [Au(3)M(2)(C triple bond CC(6)H(4)Me-p)(6)](-) (M = Cu, Ag) to provide an understanding on the electronic structure of the complexes. PMID- 17404631 TI - The chemistry of 6-hydroxyflavylium: zwitterionic base and p-quinoidal chalcones. A multiswitchable system operated by proton, electron and photon inputs. AB - The complex network of chemical reactions of the compound 6-hydroxyflavylium perchlorate was studied by different techniques including UV-vis absorption spectrophotometry, stopped-flow, electrochemistry, and photochemistry. The network is characterized by the formation of a very reactive zwitterionic base as well as by p-quinoidal chalcones. The trans-chalcone is metastable in very acidic solutions and exhibits reversible redox reactions, allowing to introduce for the first time in the flavylium network an electrochemical input. PMID- 17404632 TI - Substitution and solvent effects in the chalcones isomerization barrier of flavylium photochromic systems. AB - Useful application of photochromic compounds as optical memories implies the existence of a large kinetic barrier between the forms interconverted by light. In the case of flavylium salts, the ground state isomerization barrier between the photoisomerizable chalcone isomers is shown to correlate with the electron donating ability of the substituents, measured by their effects in the (1)H NMR chemical shifts of the aromatic protons. Substitution with electron donating groups in ring A lowers the barrier while substitution at ring B has the opposite effect. However, in water, the observed increase is higher than expected in the case of compound 4',9-dihydroxychalcone when compared with the analogous 4' dimethylamino-9-hydroxychalcone, containing a better electron donating group in the same position. Our interpretation is that the water network is providing an efficient pathway to form tautomers. In acetonitrile, unlike water, the expected order is indeed observed: E(a)(4',9-dihydroxychalcone) = 60 kJ mol(-1) < E(a) (4' dimethylamino-9-hydroxychalcone) = 69 kJ mol(-1). PMID- 17404633 TI - Resonance Raman and lifetime studies on regioselectively deuteriated ruthenium(II) polypyridyl complexes. AB - Two series of ruthenium(II) polypyridyl complexes [Ru(bipy)(2)(phpytr)](+) and [Ru(bipy)(2)(phpztr)](+) (where Hphpytr = 2-(5-phenyl-1H-[1,2,4]triazol-3-yl) pyridine and Hphpztr = 2-(5-phenyl-1H-[1,2,4]triazol-3-yl)-pyrazine) are examined by electrochemistry, UV/Vis, emission, resonance Raman, transient resonance Raman and transient absorption spectroscopy, in order to obtain a more comprehensive understanding of their excited state electronic properties. The interpretation of the results obtained is facilitated by the availability of several isotopologues of each of the complexes examined. For the pyridine-1,2,4-triazolato based complex the lowest emissive excited state is exclusively bipy based, however, for the pyrazine based complexes excited state localisation on particular ligands shows considerable solvent and pH dependency. PMID- 17404634 TI - Fast, through-bond mediated energy transfer from Ir(III) to Ru(II) in di- and tetranuclear heterometallic assemblies: elucidation of a two-step Ir --> Ir --> Ru energy transfer process. AB - Energy transfer processes triggered by light excitation (lambda(exc) = 278 nm) have been investigated in a heterometallic complex, Ir-Ru, and in a tetranuclear assembly, ((F4))(2)-Ir-Ru, containing iridium(III) and ruthenium(II) centres {Ir represents the unit [(ppy)(2)Ir(bpy)](+), Ru is [Ru(bpy)(3)](2+) and Ir(F4) is [Ir(F(2)ppy)(2)(4-bpy-C(6)H(4)-)](+) (ppy = 2-phenylpyridyl, bpy = 2,2' bipyridyl)}. In the dinuclear species, the two metallic components are linked by a biphenylene bridge connected between the 4-positions of the bpy of Ir and one of the three bpy's of Ru. The tetrametallic species comprises the dimeric unit appended with Ir(F4) units at the 4'-positions of the ppy ligands. For both the dinuclear and tetranuclear complexes, the metal centers are held at fixed distances by the interposed phenylene bridges. We have obtained the optical (absorption and emission) properties for the mononuclear species Ir(F4), Ir and Ru and for the polymetallic species Ir-Ru and ((IrF4))(2)-Ir-Ru. For Ir-Ru, the Ir --> Ru energy transfer step is exothermic by ca. 0.22 eV, based on the emission energies of the respective mononuclear components at 77 K. In turn, within ((F4))(2)-Ir-Ru, the excited state energy of Ir(F4) is ca. 0.26 eV higher than Ir. By using lambda(exc) = 278 nm, it is possible to predominantly excite the iridium-based units of both Ir-Ru and (Ir(F4))(2)-Ir-Ru. For both cases, energy transfer is found to be fast and efficient with k(en) > 2 x 10(8) s(-1), leading to detectable emission only from the Ru component both at room temperature and at 77 K. In particular, for ((F4))(2)-Ir-Ru, based on the evaluated intermetallic distances (e.g. 28.5 A for the shortest Ir(F4)-Ru distance) and the overlap integrals of donor and acceptor units, the observed energy transfer is too fast to be accounted for by through-space Forster transfer. In this species, energy transfer probably occurs in a two-step process, Ir(F4)-->Ir-->Ru, both steps involving a through-bond Dexter mechanism. PMID- 17404635 TI - Non-conjugated, phenyl assisted coupling in through bond electron transfer in a perylenemonoimide-triphenylamine system. AB - Two donor-bridge-acceptor compounds containing triphenylamine (TPA) donors and perylenemonoimide (PMI) acceptors have been studied by spectroscopic techniques and quantum chemical computation. Both systems have been observed to emit prompt and delayed fluorescence under certain conditions indicating that forward and reverse electron transfer (ET) processes can occur between the locally excited and the charge separated states. The experimental and computational results show that the TPA and PMI chromophores are better coupled by almost 50% in the meta isomers which undergo ET more readily than the para isomers. Quantum chemical calculations indicate that this unexpected situation is the result of a phenyl group on the side of the bridge being advantageously positioned in the meta isomers. This leads to more extensive delocalisation of the TPA HOMO into the bridge enhancing the total through bond electronic coupling between the TPA and PMI chromophores. The calculations also indicate a strong angle dependence of the total coupling in both isomers. The experimental results are discussed in the context of the high temperature limit of Marcus's theory of non-adiabatic ET. PMID- 17404636 TI - Self-assembly of semiconductor quantum-dots on electrodes for photoelectrochemical biosensing. AB - CdS nanoparticles linked through a duplex DNA to a Au electrode do not lead to a noticeable photocurrent upon their illumination in the presence of triethanolamine, TEOA, 20 mM, pH = 7.2. The intercalation of doxorubicin into the duplex DNA stimulates, however, the generation of a photocurrent. This is attributed to the trapping of photoexcited conduction-band electrons by the intercalator units that facilitates, by a hopping mechanism, the electron transport to the electrode. The oxidation of TEOA by valence band holes allows the formation of a steady state photocurrent. This basic phenomenon is used to probe the operation of a DNA-based machine through the assembly of CdS nanoparticles on a Au electrode. The machine includes a nucleic acid "track", (1), that binds a primer, (2), through hybridization to a predefined domain. In the presence of polymerase, the nucleotide mixture, dNTPs, and the nicking enzyme, the autonomous replication, nicking and displacement of the "waste product", (3), are activated. The "waste product" bridges the (5)-functionalized CdS nanoparticles and the nucleic acid (4)-functionalized Au electrode, resulting in the assembly of the nanoparticles on the electrode. The intercalation of doxorubicin into the DNA-CdS nanostructures results in the generation of photocurrents upon illumination in the presence of TEOA, pH = 7.2. The photocurrents are controlled by the time intervals used to operate the DNA machine. PMID- 17404637 TI - Photochemistry in everyday life: the effect of spontaneous emulsification on the photochemistry of trans-anethole. AB - The photochemical behaviour of spontaneously formed microemulsions obtained upon dilution of ethanolic solutions of trans-anethole (E-1-(4-methoxyphenyl)propene, t-A) with water is compared to that of homogeneous ethanolic t-A solutions. Significant differences in reactivity reflect the confined nature of the aggregated t-A which leads to reduced yields of isomerization and dimerization products. In contrast to homogeneous solutions, where a photostationary state enriched in the Z-isomer (c-A) is rapidly reached, the proportion of c-A formed upon irradiation of t-A microemulsions remains below 15%. In the presence of oxygen the formation of trans-anethole oxide is observed which, when formed in non-homogeneous environments, undergoes polymerization. PMID- 17404638 TI - Parameters affecting the chemical work output of a hybrid photoelectrochemical biofuel cell. AB - A hybrid photoelectrochemical biofuel cell employing the photoanode architecture of a dye-sensitized solar cell has been assembled. A porphyrin dye sensitizes a TiO(2) semiconductor over the visible range to beyond 650 nm. Photoinduced charge separation at the dye-TiO(2) interface results in electron migration to a cathode, and the holes generated on surface bound dyes oxidize soluble electron mediators. The increased [Ox] : [Red] ratio of the mediator drives the solution based enzymatic oxidation of appropriate substrates. In this report we investigate how the accumulation of anodic and cathodic products limits cell performance. The NAD(+)/NADH and benzoquinone/hydroquinone redox couples were studied as sacrificial electron donors in the absence of appropriate enzymes or substrates. Comparatively poor cell performance was observed using the benzoquinone/hydroquinone couple. This effect is explained in terms of rapid charge recombination by electron donation from the electrode to benzoquinone in solution, as compared to much less recombination with NAD(+). With the NAD(+)/NADH couple the cell performance is relatively independent of the redox poise of the anode solution, but limited by accumulation of reduction products in the cathodic compartment. Using the NAD(+)/NADH couple, the photochemical reforming of ethanol to hydrogen was demonstrated under conditions where the process would be endergonic in the dark. PMID- 17404639 TI - Site-specific electronic couplings in dyads with MLCT excited states. Intramolecular energy transfer in isomeric Ru(II)-Ru(II) cyclometalated complexes. AB - The rod-like binuclear complexes [(ttpy)Ru(tpy-ph(2)-phbpy)Ru(ttpy)](4+) and [(ttpy)Ru(tpy-ph(2)-tpy)Ru(phtbpy)](4+) (for abbreviations, see text) have been synthesized and characterized. In both complexes, the polypyridine Ru(II) centers have (N--N--N)Ru(N--N--N) and (N--N--N)Ru(C--N--N) coordination environment. The two isomeric species differ in whether the cyclometalating carbon resides on the bridging or on the terminal ligand. The two complexes have virtually identical energy levels, but MLCT excited states of different (bridging or terminal) ligand localization. They are thus ideally suited to investigate possible effects of excited-state localization on intramolecular energy transfer kinetics. In fact, ultrafast spectroscopic measurements yield different energy transfer time constants for the two isomers, with the bridge-cyclometalated complex (2.7 ps) being faster than the terminal-cyclometalated one (8.0 ps). This difference can be explained in terms of different electronic factors for Dexter energy transfer. The study highlights the peculiar intricacies of intramolecular energy transfer in inorganic dyads involving MLCT excited states. PMID- 17404640 TI - The effects of ligand substitution and deuteriation on the spectroscopic and photophysical properties of [Ru(LL)(CN)4]2- complexes. AB - The spectroscopic characterisation of a series of [Ru(LL)(CN)(4)](2-) complexes, where LL = 1,10-phenanthroline (phen) and its methyl- and phenyl-substituted derivatives and several deuteriated isotopologues are reported. The optical and vibrational properties of these complexes are compared with that of the series of 2,2'-bipyridine (bipy) derivatives and analogous [Ru(LL)(3)](2+) complexes. It has been demonstrated that substitution at the 4,4' positions of bipy and 4,7 positions of phen by electron donating (CH(3)) and withdrawing (C(6)H(5), COO(-)) groups induces a pronounced blue and red shift, respectively, in the lowest energy (1)MLCT absorption band of [Ru(LL)(CN)(4)](2-). The energy of the emission originating from the (3)MLCT excited state is found to be dependant on the nature of the vibrational modes of the aromatic rings and the electron donating and/or withdrawing properties of the substituents. Single-mode Franck-Condon analysis indicates that methyl substitution leads to a significant increase in the Huang Rhys factor (S(M)), while phenyl substitution results in a decrease in S(M) for both series (bipy and phen) of complexes. The rate of non-radiative (k(nr)) and radiative decay (k(ph)) to the ground state and the parameters of thermally activated deactivation pathways (A(4th), DeltaE(4th) and A(dd), DeltaE(dd)) were estimated from the temperature dependence of luminescence quantum yields and lifetimes. It has been demonstrated that the non-radiative decay rate and the temperature dependent decay processes are more efficient for bipy complexes than for phen derivatives due to the rigidity of the latter ligand. PMID- 17404641 TI - Highly efficient supramolecular photocatalysts for CO2 reduction using visible light. AB - We report the most efficient homogeneous photocatalyst yet for CO(2) reduction using a wide range of visible-light wavelength. We synthesized new Ru(II)-Re(I) binuclear complexes with 1,3-bis(4'-methyl-[2,2']bipyridinyl-4-yl)-propan-2-ol (bpyC3bpy) as a bridge ligand, specifically [Ru-ReP(OEt)(3)](3+) and [Ru Repy](3+) where a P(OEt)(3) or pyridine ligand coordinates on the Re site. Their photocatalytic activities were compared with [Ru-ReCl](2+), which has a Cl(-) ligand on the Re site and has recently been reported as a much better photocatalyst (Phi = 0.12, TN(CO) = 160) than a 1:1 mixed system of the corresponding Ru(II) and Re(I) mononuclear complexes. The best photocatalyst was [Ru-ReP(OEt)(3)](3+), for which Phi = 0.21 and TN(CO) = 232. A mechanistic study clearly showed that [Ru-ReP(OEt)(3)](3+) is rapidly converted into the solvento complex [Ru-ReSol](3+), (Sol = DMF or TEOA) which is the actual photocatalyst. Although similar rapid ligand substitution occurs with other supramolecules, the pyridine and Cl(-) anions accelerate the decomposition of the supramolecular photocatalysts. PMID- 17404642 TI - Binding and photochemistry of enantiomeric 2-(3-benzoylphenyl)propionic acid (ketoprofen) in the human serum albumin environment. AB - Global analysis of circular dichroism multiwavelength data and time resolved fluorescence was applied to investigate the interaction of R(-)- and S(+) ketoprofen (KP) with human serum albumin (HSA) in buffer solution at neutral pH. The most stable drug:protein adducts of 1 : 1 and 2 : 1 stoichiometry were characterized as regards the stability constants and the absolute circular dichroism spectra. The spectra of the diastereomeric 1 : 1 conjugates are negative with minima at ca. 350 nm for R(-)-KP and 330 nm for S(+)-KP, those of the 2 : 1 complexes are both negative with minimum at 340 nm and quite similar in shape to each other, thereby showing that the protein loses chiral recognition capability upon multiple binding. HSA intrinsic time resolved fluorescence data obtained exciting at 295 nm point to Trp 214 being located in the secondary binding site for both KP enantiomers. The photodegradation of the S(+)- and R(-) KP:HSA complexes was studied by steady state photolysis using lambda(irr) > 320 nm. No decrease of the photodegradation quantum yields was observed in 1 : 1 complexes. An induction time for the photodegradation course in 2 : 1 complexes was observed. Transient absorption spectroscopy at lambda(exc) = 355 nm showed that triplet KP species were formed with stereo-differentiated lifetimes and high quantum yields (0.7-0.9). Secondary transients were consistent with the occurrence of photodecarboxylation and/or photoreduction within the protein matrix. PMID- 17404643 TI - First generation TREN dendrimers functionalized with naphthyl and/or dansyl units. Ground and excited state electronic interactions and protonation effects. AB - We report the photophysical properties (absorption and emission spectra, quantum yield, and lifetime) of five dendrimers of first generation based on a TREN (tris(2-aminoethyl)amine) skeleton functionalized at the periphery with naphthyl and/or 5-dimethylamino-1-naphthalenesulfonamide (hereafter called dansyl) chromophores. Each dendrimer comprises one tertiary amine unit in the core and three branches carrying a sulfonimido unit at the periphery, each one substituted by two identical or different moieties. In particular, TD6 and TN6 contain dansyl (D) or naphthyl (N) units, respectively, while TD3B3, TN3B3 and TN3D3 contain dansyl, naphthyl or benzyl (B) units at the periphery. The spectroscopic behaviour of these dendrimers has been investigated in acetonitrile solution and compared with that of reference compounds. For all dendrimers the absorption bands are red shifted compared to those of monomeric naphthyl and dansyl reference compounds. Moreover, the intense naphthyl and dansyl fluorescence is greatly quenched because of strong interactions between the two aromatic moieties linked by a sulfonimido unit. Protonation of the amine units of the dendrimers by addition of CF(3)SO(3)H (triflic) acid causes a decrease in intensity of the luminescence and a change in the shape of the emission bands. The shapes of the titration curves depend on the dendrimer, but in any case the effect of acid can be fully reversed by successive addition of base (tributylamine). The obtained results reveal that among the intradendrimer interactions the most important one is that taking place (via mesomeric interaction) between the various chromophores and a pair of sulfonimido groups. PMID- 17404644 TI - Coumaric amide rotaxanes: effects of hydrogen bonding and mechanical interlocking on the photochemistry and photophysics. AB - Secondary amide derivatives of coumaric and ferulic acid are shown to undergo photoisomerization, forming a photostationary mixture of E- and Z-isomers. When the same chromophores are incorporated in rotaxanes, the extent of conversion to the Z-isomers is much smaller. Low temperature fluorescence experiments show that the energy barrier for non-radiative decay of the excited state is higher in the rotaxanes than in the corresponding threads, but the barriers are low in all cases. PMID- 17404645 TI - Exploiting the dithiocarbamate ligand in metal-directed self-assembly. AB - The dithiocarbamate (dtc) ligand has proved to be an extremely versatile and robust motif for metal-directed self-assembly. Its ease of formation and wide ranging coordination chemistry has led to the formation of an array of novel and complex supramolecular architectures. Well-defined structures such as macrocycles, cages, catenanes and nanodimensional assemblies can be generated using a variety of oligomeric dithiocarbamate constructs in combination with transition metals. Polymetallic assemblies containing appropriately designed host cavities have allowed the binding of cationic, anionic and neutral guest species to be investigated. The use of the dithiocarbamate ligand has recently expanded to stabilising gold nanoparticles and preparing multimetallic wires and arrays. This perspective highlights the considerable potential that this simple and versatile ligand has to offer. PMID- 17404646 TI - Glycoligands and Co(II) glycocomplexes. Investigation of the variation of the sugar-scaffold on the structure and chirality measured by circular dichroism. AB - Glycoligands using various monosaccharide platforms functionalised by three 2 picolyl groups and coordinated to Co(II) through the bidentate 2-picolyl ether moieties are interesting ligands as they efficiently induce chirality at the cobalt with a fine control of the structure through the central sugar scaffold. PMID- 17404647 TI - Stereoselective synthesis of olefins by a reductive coupling reaction. AB - Ketones and aldehydes are converted to symmetrical and (E)-olefins (1-15) by reaction with 2,4-bis(phenyl)-1,3-diselenadiphosphetane-2,4-diselenide (PhPSe(2))(2), Woollins' reagent, in refluxing toluene; use of diketones was demonstrated by the reaction of PhC(O)CH(2)C(O)Ph which gives 1,2,4,5 tetraphenylbenzene (16) in 83% yield. PMID- 17404648 TI - Tetranuclear palladium complexes with benzoquinonediimine ligands: synthesis, molecular structure and electrochemistry. AB - Rare examples of tetrapalladated derivatives in which two flat tetradentate bridging ligands are perfectly face-to-face as a result of a remarkable ancillary ligand effect are reported. PMID- 17404649 TI - Anthracene as a sensitiser for near-infrared luminescence in complexes of Nd(III), Er(III) and Yb(III): an unexpected sensitisation mechanism based on electron transfer. AB - The ligand L(1), which contains a chelating 2-(2-pyridyl)benzimidazole (PB) unit with a pendant anthacenyl group An connected via a methylene spacer, (L(1) = PB An), was used to prepare the 8-coordinate lanthanide(III) complexes [Ln(hfac)(3)(L(1))] (Ln = Nd, Gd, Er, Yb) which have been structurally characterised and all have a square antiprismatic N(2)O(6) coordination geometry. Whereas free L(1) displays typical anthracene-based fluorescence, this fluorescence is completely quenched in its complexes. The An group in L(1) acts as an antenna unit: in the complexes [Ln(hfac)(3)(L(1))] (Ln = Nd, Er, Yb) selective excitation of the anthracene results in sensitised near-infrared luminescence from the lanthanide centres with concomitant quenching of An fluorescence. Surprisingly, the anthracene fluorescence is also quenched even in the Gd(III) complex and in its Zn(II) adduct in which quenching via energy transfer to the metal centre is not possible. It is proposed that the quenching of anthracene fluorescence in coordinated L(1) arises due to intra-ligand photoinduced electron-transfer from the excited anthracene chromophore (1)An* to the coordinated PB unit generating a short-lived charge-separated state [An(.+) PB(.-)] which collapses by back electron-transfer to give (3)An*. This electron transfer step is only possible upon coordination of L(1) to the metal centre, which strongly increases the electron acceptor capability of the PB unit, such that (1)An* --> PB PET is endoergonic in free L(1) but exergonic in its complexes. Thus, rather than a conventional set of steps ((1)An* -->(3)An* --> Ln), the sensitization mechanism now includes (1)An* --> PB photoinduced electron transfer to generate charge-separated [An(.+)-PB(.-)], then back electron transfer to generate (3)An* which finally sensitises the Ln(III) centre via energy transfer. The presence of (3)An* in L(1) and its complexes is confirmed by nanosecond transient absorption studies, which have also shown that the (3)An* lifetime in the Nd(III) complex matches the rise time of Nd-centred near-infrared emission, confirming that the final step of the sequence is (3)An* --> Ln(III) energy-transfer. PMID- 17404650 TI - Sensitised near-infrared luminescence from lanthanide(III) centres using Re(I) and Pt(II) diimine complexes as energy donors in d-f dinuclear complexes based on 2,3-bis(2-pyridyl)pyrazine. AB - The luminescent transition metal complexes [Re(CO)(3)Cl(bppz)] and [Pt(CC C(6)H(4)CF(3))(2)(bppz)] [bppz = 2,3-bis(2-pyridyl)pyrazine], in which one of the diimine binding sites of the potentially bridging ligand bppz is vacant, have been used as 'complex ligands' to make heterodinuclear d-f complexes by attachment of a {Ln(dik)(3)} fragment (dik = a 1,3-diketonate) at the vacant site. When Ln = Pr, Nd, Er or Yb the lanthanide centre has low-energy f-f excited states capable of accepting energy from the (3)MLCT excited state of the Pt(II) or Re(I) centre, quenching the (3)MLCT luminescence and affording sensitised lanthanide(III)-based luminescence in the near-IR region. UV/Vis and luminescence spectroscopic titrations allowed measurement of (i) the association constants for binding of the {Ln(dik)(3)} fragment at the vacant diimine site of [Re(CO)(3)Cl(bppz)] or [Pt(CC-C(6)H(4)CF(3))(2)(bppz)], and (ii) the degree of quenching of the (3)MLCT luminescence according to the nature of the Ln(III) centre. In all cases Nd(III) was found to be the most effective of the series at quenching the (3)MLCT luminescence of the d-block component because the high density of f-f excited states of the appropriate energy make it a particularly effective energy-acceptor. PMID- 17404651 TI - Metal carbonyls as pharmaceuticals? [Ru(CO)3Cl(glycinate)], a CO-releasing molecule with an extensive aqueous solution chemistry. AB - The pharmacologically active [Ru(CO)(3)Cl(glycinate)] is shown to be in equilibrium with [Ru(CO)(2)(CO(2)H)Cl(glycinate)](-) (isomers) at around pH 3.1 which then at physiological pH reacts with more base to give [Ru(CO)(2)(CO(2))Cl(glycinate)](2-) (isomers) or [Ru(CO)(2)(CO(2)H)(OH)(glycinate)](-) (isomers). The ease with which [Ru(CO)(3)Cl(glycinate)] reacts with hydroxide results in it producing a solution in water with a pH of around 2 to 2.5 depending on concentration and making its solutions more acidic than those of acetic acid at comparable concentrations. Acidification of [Ru(CO)(3)Cl(glycinate)] with HCl gives [Ru(CO)(3)Cl(2)(NH(2)CH(2)CO(2)H)]. The crystal structures of [Ru(CO)(3)Cl(glycinate)] and [Ru(CO)(3)Cl(2)(NH(2)CH(2)CO(2)Me)] are reported. PMID- 17404652 TI - Origin of enantioselectivity in palladium-catalyzed asymmetric allylic alkylation reactions using chiral N,N-ligands with different rigidity and flexibility. AB - The chiral bidentate-N,N ligands, (S(a))-1, (S(a))-2, (S,S)-3 and (S,S)-4, were synthesized. They were shown to contain rigid 2-pyridinyl or 8-quinolinyl building blocks and the C(2)-symmetric chiral frameworks trans-2,5 dimethylpyrrolidinyl or (S)-(+)-2,2'-(2-azapropane-1,3-diyl)-1,1'-binaphthalene. In the (S(a))-2, and (S,S)-4 ligands pair, the 8-quinolinyl skeleton is directly bonded to the C(2)-symmetric chiral frameworks (S)-(+)-2,2'-(2-azapropane-1,3 diyl)-1,1'-binaphthalene or trans-2,5-dimethylpyrrolidinyl. This feature induces rigidity in this pair of ligands upon the N,N-framework. However, this does not occur for the (S(a))-1 and (S,S)-3 ligands, in which the presence of the -CH(2)- spacer between the frameworks bearing the nitrogen atom donors gives greater flexibility to the ligand. A further difference between the pairs of ligands is significant from the electronic properties of the chiral framework N-donor atom. The coordinating properties and the specific steric structural features of the (S(a))-1, (S(a))-2, (S,S)-3, and (S,S)-4 ligands are explained by their reactions with the [Pd(PhCN)(2)Cl(2)] and [Pd(eta(3)-PhCHCHCHPh)(mu-Cl)](2) substrates, in which the reported ligands form chelate complexes, with the exception of (S(a)) 2, which failed to react with [Pd(eta(3)-PhCHCHCHPh)(mu-Cl)](2). The ligands were used in the palladium-allyl catalyzed substitution reaction of 1,3-diphenylallyl acetate with dimethylmalonate, with the best result being obtained using the (S(a))-1 ligand, giving the substitution product 2-(1,3 diphenylallyl)dimethylmalonate with an enantiomeric excess of 82% in the S form and a yield of 96%. The work demonstrates that in the presence of a steric ligand control, the electronic properties of the ligand donor atoms play a role though not significant in determining the enantioselectivity of palladium(II) catalyzed allylic substitution reactions. The results of the catalytic reaction do not provide a convincing explanation considering the coordinated chiral ligand features, as rigidity or flexibility and electronic properties of the N-donor atoms. A rationalization of the results is proposed on the basis of NMR studies and DFT calculation on the cationic complexes [Pd(eta(3)-PhCHCHCHPh)(N N*)]CF(3)SO(3), (N-N* = (S(a))-1, 9; (S,S)-3, 10; (S,S)-4, 11). PMID- 17404653 TI - Iron(II) and zinc(II) monohelical binaphthyl-salen complexes with overlapping benz[a]anthryl sidearms. AB - The synthesis of the polyaromatic aldehyde 1-hydroxybenz[a]anthracene-2 carboxaldehyde is reported via a seven step protocol from 9,10-dihydroanthracene, with an overall yield of 30%. Two equivalents of the aldehyde are condensed with (R)-1,1'-binaphthyl-2,2'-diamine to produce a new binaphthyl-salen ligand, which is subsequently complexed to iron(II) and zinc(II) ions. The ligand and complexes are characterized by single-crystal X-ray crystallography. The complexes have distinct helical structures with overlapping benz[a]anthryl sidearms, and only M helices are observed. The ligand and complexes are further characterized by solution (1)H and (13)C NMR spectroscopy as well as UV-visible and ECD spectroscopies. These studies indicate that there is a single component in solution, consistent with the solid state characterization. PMID- 17404654 TI - Syntheses, X-ray structures and AACVD studies of group 11 ditelluroimidodiphosphinate complexes. AB - Reactions of Na(tmeda)[N((i)Pr(2)PTe)(2)] with CuCl, AgI or AuCl (in the presence of PPh(3)) in THF produced the coinage metal ditelluroimidodiphosphinate complexes {Cu[N((i)Pr(2)PTe)(2)]}(3), (5), {Ag[N((i)Pr(2)PTe)(2)]}(6) (6) and Au(PPh(3))[N((i)Pr(2)PTe)(2)] (7), respectively. Complexes 5, 6 and 7 were characterized in the solid state by X-ray crystallography. Complex 5 is trimeric and exhibits a highly distorted Cu(3)Te(3) ring. In contrast, the Ag(I) complex 6 is a hexamer, and forms a twelve-membered Ag(6)Te(6) ring. The replacement of the (i)Pr groups on phosphorus by Ph results in an intriguing structural change to a tetramer with a boat-shaped Ag(4)Te(4) ring in {Ag[N(Ph(2)PTe)(2)}(4).2THF (8). The gold(I) complex 7 is monomeric. Aerosol-assisted chemical vapour deposition (AACVD) of compounds 5, 6 and 7 yields CuTe, Ag(7)Te(4), AuTe(2) and Au films, respectively. The films were grown at temperatures of 300-500 degrees C and characterized by X-ray powder diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive analysis of X-rays (EDAX). PMID- 17404655 TI - Training top surgeons - or anyone else. PMID- 17404656 TI - Lessons learned from working in emergency departments in Cape Town, South Africa : a final-year medical student's perspective. PMID- 17404657 TI - Integrative and complementary medicine. PMID- 17404658 TI - This prostate cancer death should not have happened. PMID- 17404659 TI - Medicine's missing links? PMID- 17404660 TI - A refugee nurse's story.... PMID- 17404661 TI - Foreign nurses 'trickle in' - eight times slower than locals leaving. PMID- 17404662 TI - 'Just micro-size me' - tiny diabetes device to the rescue? PMID- 17404663 TI - Surprise 'social status' finding in rape study. PMID- 17404664 TI - Treatment of laryngeal lipoid proteinosis using CO2 laser. PMID- 17404665 TI - Hamilton Naki - a surgical sherpa. PMID- 17404666 TI - Iatrogenic pneumatosis intestinalis and pneumatosis hepatis. PMID- 17404667 TI - Natural involution of pulmonary hydatid cyst and an iatrogenic cause of the Cumbo sign. PMID- 17404668 TI - My TB story. PMID- 17404669 TI - High HIV/AIDS prevalence among health workers requires urgent action. PMID- 17404670 TI - Little mental disorder research in general medical journals in low - and middle income countries. PMID- 17404671 TI - Outbreak of myiasis. PMID- 17404672 TI - Prevalence of HIV infection and median CD4 counts among health care workers in South Africa. AB - OBJECTIVE: To determine the prevalence of HIV infection and the extent of disease progression based on CD4 count in a public health system workforce in southern Africa. DESIGN: Cross-sectional voluntary, anonymous, unlinked survey including an oral fluid or blood sample and a brief demographic questionnaire. SETTING: Two public hospitals in Gauteng, South Africa. SUBJECTS: All 2 032 professional and support staff employed by the two hospitals. OUTCOME MEASURES: HIV prevalence and CD4 cell count distribution. RESULTS: Overall prevalence of HIV was 11.5%. By occupation, prevalence was highest among student nurses (13.8%) and nurses (13.7%). The highest prevalence by age was in the 25 - 34-year group (15.9%). Nineteen per cent of HIV-positive participants who provided blood samples had CD4 counts less than or equal to 200 cells/ microl 28% had counts 201 - 350 cells/ microl, 18% had counts 351 - 500 cells/ microl, and 35% had counts above 500 cells/ microl. CONCLUSIONS: One out of 7 nurses and nursing students in this public sector workforce was HIV-positive. A high proportion of health care workers had CD4 counts below 350 cells/ microl, and many were already eligible for antiretroviral therapy under South African treatment guidelines. Given the short supply of nurses in South Africa, knowledge of prevalence in this workforce and provision of effective AIDS treatment are crucial for meeting future staffing needs. PMID- 17404673 TI - High prevalence of abnormal Pap smears among young women co-infected with HIV in rural South Africa - implications for cervical cancer screening policies in high HIV prevalence populations. AB - OBJECTIVE: To establish the relationship between HIV infection and cervical dysplasia in young women in rural South Africa. METHODS: This cross-sectional study was conducted at a primary health care clinic in Vulindlela, KwaZulu- Natal. Standardised questionnaires were used to collect sociodemographic and clinical presentation data from women attending family planning and other reproductive health services. Pap smears were done using standard methods. Pap smear data were linked to HIV serostatus. RESULTS: Four hundred and sixty-six women were included in the study. The median age was 24.3 years (range 15 - 55 years), and 80% were younger than 30 years. The HIV prevalence rate was 24.5% (95% confidence interval: 20.7 - 28.7%) and the prevalence of abnormal Pap smears was 16.9 - 6.4% ASCUS (atypical squamous cells of undetermined significance), 9.2% LGSIL (low-grade squamous intraepithelial lesions), and 1.3% HGSIL (high grade squamous intraepithe lial lesions). The association between HIV seropositivity and abnormal Pap results was statistically significant (p < 0.05). CONCLUSION: There is a need for more data on cervical changes in HIV co-infected women and for review of guidelines on selective Pap smear screening in high HIV prevalence settings such as sub-Saharan Africa and where access to antiretroviral treatment remains limited. PMID- 17404674 TI - Key issues in clinic functioning - a case study of two clinics. AB - OBJECTIVE: The aim of this research was to understand key issues in the functioning of two different primary care clinics serving the same community, in order to learn more about clinic management. DESIGN: An in-depth case study was conducted. A range of qualitative information was collected at both clinics. Data collected in the two clinics were compared, to gain an understanding of the important issues. SETTING: Data were collected in a government and an NGO clinic in North West province. SUBJECTS: This report presents the findings from patient and staff satisfaction surveys and in-depth individual interviews with senior staff. RESULTS: Key findings included the following: (i) there are attitudinal differences between the staff at the two clinics; (ii) the patients appreciate the services of both clinics, though they view them differently; (iii) clinic A provides a wider range of services to more people more often; (iv) clinic B presents a picture of quality of care, related to the environment and approach of staff; (v) waiting time is not as important as how patients are treated; (vi) medications are a crucial factor, in the minds of staff and patients; and (vii) a supportive, empowering organisational culture is needed to encourage staff to deliver better care to their patients. The management of the clinic is part of this culture. CONCLUSIONS: This research provides lessons regarding key issues in clinic functioning which can make a major difference to the way services are experienced. A respectful and caring approach to patients, and an organisational culture which supports and enables staff, can achieve much of this without any additional resources. PMID- 17404675 TI - Health financing reform in Kenya - assessing the social health insurance proposal. AB - Kenya has had a history of health financing policy changes since its independence in 1963. Recently, significant preparatory work was done on a new Social Health Insurance Law that, if accepted, would lead to universal health coverage in Kenya after a transition period. Questions of economic feasibility and political acceptability continue to be discussed, with stakeholders voicing concerns on design features of the new proposal submitted to the Kenyan parliament in 2004. For economic, social, political and organisational reasons a transition period will be necessary, which is likely to last more than a decade. However, important objectives such as access to health care and avoiding impoverishment due to direct health care payments should be recognised from the start so that steady progress towards effective universal coverage can be planned and achieved. PMID- 17404676 TI - The impact of allergic rhinitis on work productivity. AB - AIM: Patients with allergic rhinitis experience a multitude of symptoms that usually compromise some aspect of lifestyle. However, few data are available that specifically address the impact of allergic rhinitis on work productivity. METHODS: A questionnaire was developed and mailed to 2,065 patients enrolled in a 500,000-member managed care organisation. Patients were identified by diagnostic codes for allergic rhinitis as determined by a retrospective examination of medical and prescription claims records from January 1 2000 to December 31 2000. Patients were divided into three different care groups according to whether they were managed by family physicians, by allergists, or were self-managed. RESULTS: Chi-square and analysis of variance tests revealed significant differences among the three care groups (p<0.05) for years with allergies, symptoms, family history, testing, immunotherapy, test value, and prescribed antihistamine use. Multiple linear regression analysis revealed that sleep, health, certain allergy symptoms and prescribed antihistamines were significantly related to work productivity. CONCLUSIONS: The results of this study revealed that the ability of individuals with allergic rhinitis to engage in productive work is influenced by sleep, health-related quality of life (HRQoL), specific symptoms, and prescribed antihistamine use. PMID- 17404677 TI - Non-participation may bias the results of a psychiatric survey: an analysis from the survey including magnetic resonance imaging within the Northern Finland 1966 Birth Cohort. AB - OBJECTIVE: A major reason for limited validity of research is non-participation. Subjects with severe mental illness tend to cumulate in the group of non participants, causing selection bias. We tested the hypothesis that severe psychosis is linked to non-participation in a field survey including magnetic resonance imaging (MRI) of the brain, psychiatric interviews and cognitive testing among subjects with psychosis. Furthermore, we wanted to explore other associative factors expected to affect non-participation. METHODS: Members of the Northern Finland 1966 Birth Cohort with a lifetime diagnosis of psychosis (N = 145) were invited to participate in the survey conducted in 1999-2001. Non participation was determined by refusal or loss of contact. Data were gathered in earlier phases of the follow-up study or using register data. RESULTS: Ninety-one (63%) subjects attended the study. Compared to participants, non-participants were more often patients with schizophrenia and had more psychiatric hospitalisations, they had more positive psychosis symptoms during their illness course and they were more often on disability pension. Married subjects participated more often than those who were not married. CONCLUSION: This study suggests that among subjects with psychosis, particularly those subjects who have the most severe course of illness are less willing to participate. This may lead to biased estimates when studying subjects with severe mental disorders. PMID- 17404678 TI - Transvaginal repair of ureterovaginal fistula by Latzko technique. AB - Ureterovaginal fistula is an uncommon but serious sequela of unrecognized distal ureteral injury during pelvic operations. Traditionally, it is managed either by endoscopic internal ureteral stenting or by ureteral reimplantation. We report a case of ureterovaginal fistula that failed to respond to ureteral stenting. Because the patient had a history of several laparotomies and intra-abdominal abscesses, she was at high risk for complications with a transabdominal operation. Therefore, we used transvaginal Latzko partial colpocleisis and successfully resolved the ureterovaginal fistula. PMID- 17404679 TI - Is there an association between depressive and urinary symptoms during and after pregnancy? AB - Depressive symptoms and urinary symptoms are both highly prevalent in pregnancy. In the general population, an association is reported between urinary symptoms and depressive symptoms. The association of depressive and urinary symptoms has not yet been assessed in pregnancy. In this study, we assessed (1) the prevalence of depressive symptoms, over-active bladder (OAB) syndrome, urge urinary incontinence (UUI) and stress urinary incontinence (SUI) during and after pregnancy using the Center for Epidemiologic Studies Depression Scale (CES-D) and the Urogenital Distress Inventory (UDI) and (2) the association of depressive symptoms with urinary incontinence and over-active bladder syndrome during and after pregnancy, controlling for confounding socioeconomic, psychosocial, behavioural and biomedical factors in a cohort of healthy nulliparous women. Our data show a significant increase in prevalence of depressive symptoms, UUI, SUI and OAB during pregnancy and a significant reduction in prevalence of depressive symptoms, SUI and OAB after childbirth. UUI prevalence did not significantly decrease after childbirth. In univariate analysis, urinary incontinence and the OAB syndrome were significantly associated with a CES-D score indicative of a possible clinical depression at 36 weeks gestation. However, after adjusting for possible confounding factors, only the OAB syndrome remained significantly associated (OR 4.4 [1.8-10.5]). No association was found between depressive and urinary symptoms at 1 year post-partum. Only OAB was independently associated with depressive symptoms during pregnancy. Possible explanations for this association are discussed. PMID- 17404680 TI - Actinomyces meyeri osteomyelitis of the symphysis pubis following pubovaginal sling. AB - Previous reports of pubic osteomyelitis associated with bone anchors describe symptoms presenting 10 days to 12 months postoperatively with typical pathogens including Streptococcus, Staphylococcus, Citrobacter, and Pseudomonas (Fitzgerald et al., Int Urogynecol J 10:346-348, 1999 Enzler et al., J Bone Jt Surg Am 81 A(12):1736-1740, 1999; Graham and Dmochowski, J Urol 168:2055-2058, 2004; Fialkow et al., Urology 64(6):1127-1132, 2004; Matkov et al., J Urol 160(4):1427, 1998). This case illustrates the potential for protracted symptoms and delayed diagnosis of pubic osteomyelitis with the fastidious pathogen of Actinomyces meyeri. PMID- 17404681 TI - Publication ethics: science at its best and always informative. PMID- 17404682 TI - Intracardiac thrombi in pediatric patients: presentation profiles and clinical outcomes. AB - We document the presentation profiles, treatment strategies, and clinical outcomes in a relatively large cohort of pediatric patients with intracardiac thrombi (ICT). We performed a retrospective review of patients diagnosed with ICT by echocardiography at a tertiary pediatric hospital during a 10-year period. These patients received medical therapy or thrombectomy. We provided echocardiographic descriptions of the ICT-size, chamber location, and mobility/morphology. The outcome measures were ICT (persistence, resolution, or embolization), effectiveness of therapy, and patient morbidity and mortality. There were 40 ICT diagnosed in 31 patients (22 males and 9 females). Mean age at diagnosis was 8.8 years (range, 15 days to 18 years). Overall mortality was 12/31 patients (39%); only one death was attributed to ICT embolization. Embolic events occurred in 4/31 patients (13%). The most common initial therapies included heparin infusion (n = 15), warfarin (n = 7), and aspirin (n = 7). The ICT resolved with medical therapy alone in 19/30 patients (63%). One patient required surgical thrombectomy. The cohort was divided into group 1 (dilated cardiomyopathy), group 2 (status post Fontan operation), and group 3 (other diagnoses). In group 1 (n = 11), there were 8 deaths. Embolization occurred in 2/5 large ICT, resulting in cerebral infarction and death (n = 1) and renal infarction (n = 1). The most common ICT location was the left ventricle (n = 10). Severe ventricular systolic dysfunction was present in 10/11 patients (91%). In group 2 (n = 9), there was 1 death. Embolization occurred in 1/7 large ICT, resulting in seizures and temporary paresis. All ICT were located in the Fontan pathway. Severe ventricular systolic dysfunction was present in 2/9 patients (22%). In group 3 (n = 11), there were 3 deaths. Embolization occurred in 1/9 small ICT, resulting in coronary emboli. ICT are most commonly diagnosed in pediatric patients with dilated cardiomyopathy or patients status post Fontan operation. The majority of ICT resolve with medical therapy. Larger ICT tend to embolize more frequently, and the morbidity secondary to embolization is significant. Rarely is mortality due to ICT embolization. The prognosis is poor for patients with left ventricular ICT or ICT in the presence of ventricular systolic dysfunction. PMID- 17404683 TI - Solitary thoracic intradural extramedullary plasmacytoma. AB - The bodies of the vertebrae are common locations for plasma cell diseases such as multiple myeloma and solitary plasmacytoma. Secondary invasion of the epidural space is infrequent but can cause neurological symptoms. Spinal cord compression due to pure intradural plasma cell infiltration is very rare. The authors report a 25-year-old woman who developed a progressive difficulty in walking due to a solitary spinal dural plasmacytoma. This is the first reported example in the English language literature of a purely intradural spinal plasmacytoma in a patient without other myelomatous lesions. An entirely intradural solitary plasmacytoma has a relatively better prognosis. PMID- 17404684 TI - Delayed massive epistaxis following endonasal transsphenoidal surgery. AB - OBJECTIVE: To describe delayed massive epistaxis, a rare but serious complication after transsphenoidal surgery. PATIENTS: Two patients underwent microscopic transsphenoidal surgery through a right endonasal approach for nonfunctioning pituitary adenoma. Severe epistaxis suddenly developed on day 8 and on day 13, respectively, after uneventful surgery. In each patient, the epistaxis was due to arterial haemorrhage from the left posterior nasal cavity. Otolarygologists failed to detect the origin of bleeding which was refractory to conservative treatment. Emergency internal carotid or external carotid angiography, performed during tight packing of the posterior nasal cavity, did not show abnormal findings; nevertheless, each patient was treated successfully by endovascular embolisation of the external carotid artery without further complication. CONCLUSION: When severe delayed epistaxis follows transsphenoidal surgery and damage to the internal carotid artery has been ruled out, endovascular embolisation of the external carotid artery should be considered in patients refractory to conservative treatment. PMID- 17404685 TI - Single amino acid substitutions in the HSV-1 helicase protein that confer resistance to the helicase-primase inhibitor BAY 57-1293 are associated with increased or decreased virus growth characteristics in tissue culture. AB - Two mutants (BAYr1 and BAYr2) that are 100-fold and >3000-fold resistant, respectively, to the helicase-primase inhibitor (HPI) BAY 57-1293 were derived from a plaque-pure parental strain, HSV-1 SC16 cl-2. BAYr1 has two substitutions in the HSV-1 helicase (UL5) protein (A4 to V; K356 to Q) and BAYr2 has one (G352 to R). It was shown reproducibly that BAYr1 grows to higher titres in tissue culture while BAYr2 grows more slowly than wild-type. Marker transfer experiments confirmed that K356Q and G352R are the drug-resistance mutations and that they are directly associated with differences in virus growth in tissue culture. When BAYr1 was tested in a murine infection model, this virus was shown to be fully pathogenic. We present evidence that single mutations close to a predicted functional domain of an essential HSV-1 replication enzyme (helicase) are associated with drug resistance and virus growth characteristics. PMID- 17404686 TI - Induced expression of oryzain alpha gene encoding a cysteine proteinase under stress conditions. AB - Oryzain alpha-A, a cysteine proteinase gene was cloned from rice (Oryza sativa L. cv. Aichi-asahi) leaves infected with Magnaporthe grisea. The protein sequence deduced for oryzain alpha-A shares high identity with that of oryzain alpha, a gene expressed in germinating rice seed. Oryzain alpha-A gene expression was induced by the blast fungus, Magnaporthe grisea, and the transcript level was even higher in the compatible interaction with rice than in the incompatible interaction. Expression of oryzain alpha-A was also inducible by wounding, ultraviolet radiation, and treatment with salicylic acid and abscisic acid, with no expression induced by methyl jasmonate. The function of oryzain alpha-A in cell death in rice is discussed. PMID- 17404687 TI - Morphological variation in leaf shape in Ainsliaea apiculata with special reference to the endemic characters of populations on Yakushima Island, Japan. AB - We analyzed leaf shape variations in Ainsliaea apiculata Sch. Bip. to evaluate the uniqueness of morphological characters in populations on Yakushima Island, Kagoshima Prefecture, Japan. Leaf size and shape from populations on Yakushima Island (n = 300) were compared with those from populations in other areas of Japan (n = 300). A considerable amount of variation occurred in leaf size in A. apiculata populations both on Yakushima Island and elsewhere, but clear discontinuities in leaf size were not detected. Some variants previously thought to be endemic to Yakushima Island, i.e., A. apiculata var. acerifolia and A. apiculata var. rotundifolia, were also found in other locations in Japan. Moreover, these leaf types were found to be continuous with the typical leaf shape of A. apiculata var. apiculata via various intermediate types, suggesting the need for future revision of these taxa. Based on these results, we reevaluated the uniqueness of the Yakushima populations of A. apiculata in terms of leaf variation. The uniqueness of the Yakushima populations was defined by a more diverse leaf shape than found in populations from other areas. PMID- 17404688 TI - Progesterone regulation of implantation-related genes: new insights into the role of oestrogen. AB - Genomic profiling was performed on explants of late proliferative phase human endometrium after 24-h treatment with progesterone (P) or oestradiol and progesterone (17beta-E(2)+P) and on explants of menstrual phase endometrium treated with 17beta-E(2)+P. Gene expression was validated with real-time PCR in the samples used for the arrays, in endometrium collected from early and mid secretory phase endometrium, and in additional experiments performed on new samples collected in the menstrual and late proliferative phase. The results show that late proliferative phase human endometrium is more responsive to progestins than menstrual phase endometrium, that the expression of several genes associated with embryo implantation (i.e. thrombomodulin, monoamine oxidase A, SPARC-like 1) can be induced by P in vitro, and that genes that are fully dependent on the continuous presence of 17beta-E(2) during P exposure can be distinguished from those that are P-dependent to a lesser extent. Therefore, 17beta-E(2) selectively primes implantation-related genes for the effects of P. PMID- 17404689 TI - Combining immune cell and viral therapy for the treatment of cancer. AB - A variety of viral-based and immune cell therapies have been proposed for use in the treatment of cancer. One possible approach to improve the effectiveness of these biological agents may be to combine them such that we can take advantage of natural immune cell-pathogen relationships. Here we discuss these potential approaches with particular emphasis on the use of immune cells as carrier vehicles to deliver viral therapies to the tumor. PMID- 17404690 TI - Relation between impaired antiplatelet response to clopidogrel and possible pleiotropic effects. AB - BACKGROUND: The study was designed to determine whether impaired antiplatelet response to clopidogrel but not to aspirin may be responsible for loss of pleiotropic effects of the drug. METHODS: Study included 34 consecutive patients with STEMI undergoing primary percutaneous coronary intervention (PCI) with stent implantation treated with aspirin (loading dose 300 mg followed by 75 mg/day) and clopidogrel (loading dose 600 mg followed by 75 mg/day). On the basis of Platelet Function Analyzer (PFA)-100 test which measured closure times (CT) in test with collagen/epinephrine (CEPI-CT) or collagen/adenosine diphosphate (CADP-CT) patients were stratified after 7 days from admission as full aspirin or clopidogrel responders (CEPI-CT or CADP-CT = 300 sec., respectively) and non-full aspirin or clopidogrel responders (CEPI-CT or CADP-CT < 300 sec., respectively). High sensitivity C-reactive protein (hs-CRP) was measured at baseline and after 7 days of treatment. RESULTS: All patients received comparable statin treatment. Median and interquartile ranges (IQR) of hs-CRP increased significantly from 2.5 mg/L (0.4-44.8) at baseline to 8.05 mg/L (1.4-33.9) at day 7 (P = .002) in non full clopidogrel responders subgroup and only slightly in the full clopidogrel responders subgroup (2.45 mg/L, IQR 0.4-48.3 vs. 4.2 mg/L, IQR 1.9-17.5) (P = .3) remaining within reference intervals. On the contrary median and IQR of hs-CRP increased significantly in both non-full aspirin responders (2.4 mg/L, IQR 1.3 3.3 vs. 5.8 mg/L, IQR 3.2-14.8, P = .01) and full aspirin responders (2.9 mg/L, IQR 2.0-3.7 vs. 5.6 mg/L, IQR 4.3-12.9, P = .04). CONCLUSIONS: Impaired antiplatelet response to clopidogrel but not to aspirin may contribute to smaller anti-inflammatory response in patients with ST-elevation myocardial infarction. PMID- 17404691 TI - Hypoxia and adaptive landscapes in the evolution of carcinogenesis. AB - Conceptual models of epithelial carcinogenesis typically depict a sequence of heritable changes that give rise to a population of cells possessing the hallmarks of invasive cancer. We propose the evolutionary dynamics that give rise to the phenotypic properties of malignant cells must be understood within the context of specific selection forces generated by the microenvironment. This can be accomplished by using an "inverse problem" approach in which we use observed typical phenotypic traits of primary and metastatic cancers to infer the evolutionary dynamics. This has led to the hypothesis that heritable changes in genes controlling cellular proliferation, apoptosis, and senescence, while necessary, are not usually sufficient to produce an invasive cancer. In addition to these evolutionary steps, we propose that the common observation of aerobic glycolysis in human cancers indicates, via the inverse problem analysis, that adaptation to hypoxia and acidosis must be a major component of the carcinogenic sequence. The details of the hypothesis are based on recognition that premalignant populations evolve within ducts and remain separated from their blood supply by a basement membrane. As tumor cells proliferate into the lumen, diffusion-reaction kinetics enforced by this separation result in hypoxia and acidosis in regions of the tumor the most distant from the basement membrane. This produces new evolutionary selection forces that promote constitutive upregulation of glycolysis and resistance to acid-induced toxicity. We hypothesize that these phenotypic adaptations are critical late steps in carcinogenesis conferring proliferative advantages even in normoxic conditions by allowing the population to produce an acidic environment (through aerobic glycolysis) which is toxic to other local cell populations and promotes extracellular matrix degradation, increasing invasiveness. PMID- 17404692 TI - Hypoxia and cancer. PMID- 17404693 TI - Hypoxia-dependent anti-inflammatory pathways in protection of cancerous tissues. AB - The evolutionarily selected tissue-protecting mechanisms are likely to be triggered by an event of universal significance for all surrounding cells. Such an event could be damage to blood vessels, which would result in local tissue hypoxia. It is now recognized that tissue hypoxia can initiate the tissue protecting mechanism mediated by at least two different biochemical pathways. The central message of this review is that tumor cells are protected from immune damage in hypoxic and immunosuppressive tumor microenvironments due to the inactivation of anti-tumor T cells by the combined action of these two hypoxia driven mechanisms. Firstly, tumor hypoxia-produced extracellular adenosine inhibits anti-tumor T cells via their G(s)-protein-coupled and cAMP-elevating A2A and A2B adenosine receptors (A2AR/A2BR). Levels of extracellular adenosine are increased in tumor microenvironments due to the changes in activities of enzymes involved in adenosine metabolism. Secondly, TCR-activated and/or tumor hypoxia exposed anti-tumor T cells may be inhibited in tumor microenvironments by Hypoxia inducible Factor 1alpha (HIF-1alpha) Hence, HIF-1alpha activity in T cells may contribute to the tumor-protecting immunosuppressive effects of tumor hypoxia. Here, we summarize the data that support the view that protection of hypoxic cancerous tissues from anti-tumor T cells is mediated by the same mechanism that protects normal tissues from the excessive collateral damage by overactive immune cells during acute inflammation. PMID- 17404694 TI - Interaction between cadmium, lead and potassium fertilizer (K2SO4) in a soil plant system. AB - A pot experiment was conducted to examine the influence of potassium (K) fertilizer (K(2)SO(4)) application on the phytoavailability and speciation distribution of cadmium (Cd) and lead (Pb) in soil. Spring wheat (Triticum aestivum L.) was selected as the test plant. There were seven treatments including single and combined contamination of Cd and Pb. CdCl(2).2.5 H(2)O and Pb(NO(3))(2) were added to the soil at the following dosages: Cd + Pb = 0.00 + 0.00, 5.00 + 0.00, 25.0 + 0.00, 0.00 + 500, 0.00 + 1000, 5.00 + 500 and 25.0 + 1000 mg kg(-1), denoted by CK, T1, T2, T3, T4, T5 and T6, respectively. The K fertilizer had five levels: 0.00, 50.0, 100, 200 and 400 mg K(2)O kg(-1) soil, denoted by K0, K1, K2, K3 and K4, respectively. The results showed that the K fertilizer promoted the dry weight (DW) of wheat in all treatments and alleviated the contamination by Cd and Pb. The application of K(2)SO(4) reduced the uptake of Cd in different parts including roots, haulms and grains of wheat; the optimum dosage was the K2 level. K supply resulted in a significant (P < 0.05) decrease in the soluble plus exchangeable (SE) fraction of Cd and there was a negative correlation (not significant, P > 0.05) between the levels of K and the SE fraction of Cd in soil. The application of the K fertilizer could obviously restrain the uptake of Pb by wheat and there were significant (P < 0.05) negative correlations between the concentrations of Pb in grains and the levels of K in soil. K supply resulted in a decrease in the SE fraction of Pb (except the K1 level) from the K0 to K4 levels. At the same time, the application of the K fertilizer induced a significant (P < 0.05) decrease in the weakly specifically adsorbed (WSA) fraction of Pb and a significant (P < 0.05) increase in the bound to Fe-Mn oxides (OX) fraction of Pb. At different K levels, the concentration of Pb in the roots, haulms and grains had a positive correlation with the SE (not significant, P > 0.05) and WSA (significant, P < 0.05) fractions of Pb in the soil. All the K application levels in this experiment reduced the phytoavailability of Cd and Pb. Thus, it is feasible to apply K fertilizer (K(2)SO(4)) to alleviate contamination by Cd and/or Pb in soil. Moreover, the level of K application should be considered to obtain an optimal effect with the minimum dosage. PMID- 17404695 TI - Bacterial growth yields on EDTA, NTA, and their biodegradation intermediates. AB - Ethylenediaminetetraacetic acid (EDTA) and nitrilotriacetic acid (NTA) are widely used anthropogenic chelating agents for control of metal speciation and are ubiquitous in natural waters and wastewaters. This is the first report of systematic measurement of the growth yields of a mixed culture (BNC1-BNC2) on EDTA and its biodegradation intermediates, and of Aminobacter aminovorans (aka Chelatobacter heintzii) ATCC 29600 on NTA and its biodegradation intermediates. The yields measured for BNC1-BNC2 co-culture were 75.0 g of cell dry weight (CDW) (mole of EDTA)(-1), 68.6 g of CDW (mole of ED3 A)(-1), 51.2 g of CDW (mole of N,N'-EDDA)(-1), 34.5 g of CDW (mole of ED)(-1), 26.3 g of CDW (mole of IDA)(-1), 12.2 g of CDW (mole of glycine)(-1), and 9.7 g of CDW (mole of glyoxylate)(-1). The yields measured for A. aminovorans were 44.3 g of CDW (mole of NTA)(-1), 37.9 g of CDW (mole of IDA)(-1), 15.2 g of CDW (mole of glycine)(-1), and 10.4 g of CDW (mole of glyoxylate)(-1). The biodegradation pathways of EDTA, NTA, and several of their metabolic intermediates include reactions catalyzed by oxygenase enzymes, which may reduce energy available for cell synthesis. Comparison of measured yields with predicted yields indicates that the effect of oxygenase reaction on cell yield can be quantified experimentally as well as modeled based on thermodynamics. PMID- 17404696 TI - Digestion of sand-laden manure slurry in an upflow anaerobic solids removal (UASR) digester. AB - Studies on the performance of a laboratory scale upflow anaerobic solids removal (UASR) digester were carried out using sand-laden cow manure slurries having total solids (TS) concentration as 50 and 100 g/l. Hydraulic retention time (HRT) was maintained as 32.4 days, which resulted in the volatile solids (VS) loading rates of 1 and 1.64 g/l d. The UASR system was designed to remove sand from the manure slurry, while anaerobically digesting biodegradable solids inside a single reactor. To enhance the contact of microorganisms and substrate, the liquor from the top of the digester was recirculated through the bed of settled solids at its bottom. Volatile solids reduction through this process was observed to be 62% and 68% in the case of feed slurries having TS concentration as 50 and 100 g/l (referred in the text as 5% and 10% feed slurries), respectively. The methane production rates were observed to be 0.22 and 0.38 l/l d, while methane yield was 0.21 and 0.27 l CH(4)/g VS loaded, for 5% and 10% feed slurries, respectively. This indicates that the increase in the VS loading had a positive impact on methane production rate and methane yield. It would be of interest to study the performance of a UASR digester at higher solids loadings and with longer solids retention times. Nonetheless, the presented study showed that sand-laden manure slurries can be successfully digested in a UASR digester producing methane energy equivalent to 4 kW h per m(3) of digester volume per day. PMID- 17404697 TI - [The problems with parallel narcosis. Professional and legal limits of delegation of anaesthesiological responsibilities to non-medical personnel]. AB - The increasing mechanisation, specialisation and sub-specialisation in medicine have enduringly supported the delegation of originally medical activities to non medical personnel and sometimes also made it necessary. Economical considerations have recently given additional impulse to these developments. It is indisputable that medical activities can be delegated to assistant personnel, however, it is equally indisputable that within the scope of the total spectrum of medical activities, there are limits to the extent of delegation, i.e. activities reserved exclusively for medical doctors. These include, by consensus of opinion, the physical examination, diagnosis, assessment of indication, determination of the therapy plan and informing the patient. The following article justifies from professional and legal viewpoints why anaesthesia also belongs to the genuine medical duties and is reserved exclusively for medical personnel. Therefore, the correct performance of parallel narcosis is coupled with far-reaching liability risks for all participants involved in this form of organisation or those responsible for them. PMID- 17404698 TI - Mechanisms of tumour cell invasion and metastasis. PMID- 17404699 TI - Long-term cost-effectiveness of donepezil for the treatment of Alzheimer's disease. AB - BACKGROUND: (Acetyl-)cholinesterase (ChE) inhibitors have been approved for the treatment of mild to moderate Alzheimer's disease (AD). However, use of ChE inhibitors is limited by budget constraints and disincentives on the side of health insurances and nursing care insurances. OBJECTIVE: To analyse under what conditions the application of the acetylcholinesterase inhibitor donepezil is favourable for the treatment of patients with AD from the perspective of health insurance and nursing care insurance companies in Germany, taking into account factors such as start and duration of treatment, duration of follow-up, drug costs, internalization of opportunity costs and varying mortality and efficacy rates. METHODS: Transition probabilities from a Swedish study and German cost data for donepezil were merged in a Markov model to follow a cohort of patients over a period of 5-10 years. We defined a base case with 1 year treatment and follow-up over 5 years and varied treatment length, follow-up interval and cost factors in sensitivity analyses. RESULTS: In the base case, the ChE inhibitor donepezil did not lead to cost savings but to a cost-effective outcome on side of health insurances and nursing care insurances. Early treatment of AD and internalization of opportunity costs (caring time devoted to patients) led to less costs per quality adjusted life years gained. However, results are very sensitive with respect to varying mortality and efficacy rates. CONCLUSION: The application of donepezil may be cost-effective, but considerable uncertainties remain. Moreover, the way the reimbursement system in Germany is presently arranged does not support the application of ChE inhibitors. PMID- 17404700 TI - [Vacuum-assisted closure therapy and wound coverage in soft tissue injury. Clinical use]. AB - This study presents the results of a meta-analysis based on the literature dealing with the clinical applications of vacuum-assisted closure (VAC) in the areas of chronic wounds, acute posttraumatic wounds, compartment syndromes, or injuries of the upper extremities. The studies were analysed for validity, significance of conclusion with respect to success rate, publications, and economic efficacy. The data show that with VAC a very valuable technique has been added to an integrated therapeutic concept of soft tissue reconstruction. However, clinical data from prospective randomised trials to support some of the positive aspects seen in the daily clinical application of the technique are still missing. These would create a sound basis demonstrating the economic efficacy of the technique. PMID- 17404701 TI - A strategy on constructing core collections by least distance stepwise sampling. AB - A strategy was proposed for constructing core collections by least distance stepwise sampling (LDSS) based on genotypic values. In each procedure of cluster, the sampling is performed in the subgroup with the least distance in the dendrogram during constructing a core collection. Mean difference percentage (MD), variance difference percentage (VD), coincidence rate of range (CR) and variable rate of coefficient of variation (VR) were used to evaluate the representativeness of core collections constructed by this strategy. A cotton germplasm collection of 1,547 accessions with 18 quantitative traits was used to construct core collections. Genotypic values of all quantitative traits of the cotton collection were unbiasedly predicted based on mixed linear model approach. By three sampling percentages (10, 20 and 30%), four genetic distances (city block distance, Euclidean distance, standardized Euclidean distance and Mahalanobis distance) combining four hierarchical cluster methods (nearest distance method, furthest distance method, unweighted pair-group average method and Ward's method) were adopted to evaluate the property of this strategy. Simulations were conducted in order to draw consistent, stable and reproducible results. The principal components analysis was performed to validate this strategy. The results showed that core collections constructed by LDSS strategy had a good representativeness of the initial collection. As compared to the control strategy (stepwise clusters with random sampling strategy), LDSS strategy could construct more representative core collections. For LDSS strategy, cluster methods did not need to be considered because all hierarchical cluster methods could give same results completely. The results also suggested that standardized Euclidean distance was an appropriate genetic distance for constructing core collections in this strategy. PMID- 17404702 TI - Variation in natriuretic peptides and mitral flow indexes during successful ventilatory weaning: a preliminary study. AB - OBJECTIVE: To assess the cardiac consequences of successful respiratory weaning using the variations of circulating B-type and atrial natriuretic peptides (BNP, ANP) and Doppler mitral flow. DESIGN: A prospective preliminary observational study. SETTING: A 14-bed medical ICU in a French university hospital. PATIENTS: Thirty-one patients undergoing a spontaneous breathing trial on a T-tube. INTERVENTIONS: Circulating BNP and ANP levels and Doppler-derived E/A ratio and deceleration time of the E wave were measured before and 1 h after disconnection. RESULTS: BNP levels increased from 299 pg/ml (range 56-1079) to 412 pg/ml (147 1324) (p=0.02) in patients with systolic left ventricular dysfunction, decreased from 98 pg/ml (25-337) to 45 pg/ml (38-180) (p=0.04) in patients with right ventricular dilation and remained unchanged in patients with neither of these cardiac abnormalities. Overall ANP levels increased from 33 pg/ml to 67 pg/ml (p<0.001) regardless of ventricular function. The E/A ratio increased from 0.91 (0.66-3.56) to 1.17 (0.5-4.76), (p=0.01), after disconnection, whereas deceleration time of E wave decreased from 185 ms (120-280) to 160 ms (70-206) (p=0.02). CONCLUSION: During successful weaning from mechanical ventilation ANP levels increase in all patients whereas changes in BNP levels depend on underlying cardiac function. Changes in Doppler mitral flow indexes following ventilator disconnection suggest an increase in left-ventricular filling pressure. PMID- 17404703 TI - Should elderly patients be admitted to the intensive care unit? AB - As the general population ages, an increasing number of patients over 80 years are being admitted to the intensive care unit (ICU). Selection of older patients for ICU admission results in lower rates of co-morbidities and underlying fatal diseases. After adjustment for disease severity, ICU and post-ICU mortality rates are higher in elderly patients than in younger populations. Age itself explains only a small part of the increased hospital mortality, suggesting that specific information such as functional, cognitive, and nutritional status, as well as co morbidities, should be collected to predict mortality in elderly ICU patients. The long-term prognosis depends chiefly on functional status, whereas initial disease severity no longer influences mortality. According to our review, it is impossible to define evidence-based recommendations for ICU admission of the elderly. This justifies further studies that encompass several aspects, such as the initial triage process and the long-term prognosis (mortality, autonomy and quality of life). PMID- 17404704 TI - Subsyndromal delirium in the ICU: evidence for a disease spectrum. AB - OBJECTIVE: ICU delirium is common and adverse. The Intensive Care Delirium Screening Checklist (ICDSC) score ranges from 0 to 8, with a score of 4 or higher indicating clinical delirium. We investigated whether lower (subsyndromal) values affect outcome. PATIENTS: 600 patients were evaluated with the ICDSC every 8[Symbol: see text]h. MEASUREMENTS AND RESULTS: Of 558 assessed patients 537 noncomatose patients were divided into three groups: no delirium (score = 0; n = 169, 31.5%), subsyndromal delirium (score = 1-3; n = 179, 33.3%), and clinical delirium (score >or=4; n = 189, 35.2%). ICU mortality rates were 2.4%, 10.6%, and 15.9% in these three groups, respectively. Post-ICU mortality was significantly greater in the clinical delirium vs. no delirium groups (hazard ratio = 1.67) after adjusting for age, APACHE II score, and medication-induced coma. Relative ICU length of stay was: no delirium < subsyndromal delirium < clinical delirium and hospital LOS: no delirium < subsyndromal delirium approximately clinical delirium. Patients with no delirium were more likely to be discharged home and less likely to need convalescence or long-term care than those with subsyndromal delirium or clinical delirium. ICDSC score increments higher than 4/8 were not associated with a change in mortality or LOS. CONCLUSIONS: Clinical delirium is common, important and adverse in the critically ill. A graded diagnostic scale permits detection of a category of subsyndromal delirium which occurs in many ICU patients, and which is associated with adverse outcome. PMID- 17404706 TI - Comment on Vieillard-Baron et al.: "Bedside echocardiographic evaluation of hemodynamics in sepsis: is qualitative evaluation sufficient?". PMID- 17404707 TI - Comment on Camus et al.: "Molecular adsorbent recirculating system dialysis in patients with acute liver failure who are assessed for liver transplantation". PMID- 17404709 TI - Potential drug sequestration during extracorporeal membrane oxygenation: results from an ex vivo experiment. AB - OBJECTIVE: Using an ex vivo simulation model we set out to estimate the amount of drug lost due to sequestration within the extracorporeal circuit over time. DESIGN: Simulated closed-loop extracorporeal membrane oxygenation (ECMO) circuits were prepared using a 1.5-m2 silicone membrane oxygenator. Group A consisted of heparin, dopamine, ampicillin, vancomycin, phenobarbital and fentanyl. Group B consisted of epinephrine, cefazolin, hydrocortisone, fosphenytoin and morphine. Drugs were tested in crystalloid and blood-primed circuits. After administration of a one-time dose of drugs in the priming fluid, baseline drug concentrations were obtained (P0). A simultaneous specimen was stored for stability testing at 24 h (P4). Serial post-membrane drug concentrations were then obtained at 30 min (P1), 3 h (P2) and 24 h (P3) from circuit fluid. MEASUREMENTS AND RESULTS: One hundred and one samples were analyzed. At the end of 24 h in crystalloid-primed circuits, 71.8% of ampicillin, 96.7% of epinephrine, 17.6% of fosphenytoin, 33.3% of heparin, 17.5% of morphine and 87% of fentanyl was lost. At the end of 24 h in blood-primed extracorporeal circuits, 15.4% of ampicillin, 21% of cefazolin, 71% of voriconazole, 31.4% of fosphenytoin, 53.3% of heparin and 100% of fentanyl was lost. There was a significant decrease in overall drug concentrations from 30 min to 24 h for both crystalloid-primed circuits (p = 0.023) and blood-primed circuits (p = 0.04). CONCLUSIONS: Our ex vivo study demonstrates serial losses of several drugs commonly used during ECMO therapy. Therapeutic concentrations of fentanyl, voriconazole, antimicrobials and heparin cannot be guaranteed in patients on ECMO. PMID- 17404710 TI - A role for cyclooxygenase in aging-related changes of beta-adrenoceptor-mediated relaxation in rat aortas. AB - beta-Adrenoceptor-mediated vasorelaxation decreases with age in various vascular beds. The present study investigated the roles of cyclooxygenase (COX) on beta adrenoceptor vasorelaxation by isoprenaline in 8- and 54-week-old rat aortas. The vasorelaxation responses by isoprenaline (0.03-3 microM) were significantly reduced in 54-week-old aortas compared to 8-week. Addition of the non-selective COX inhibitors indomethacin (10 microM) or aspirin (10 microM) restored isoprenaline vasorelaxation of 54-week-old aortas to levels found in 8-week-old aortas. This suggests the involvement of COX prostanoids in the age-related reduction of beta-adrenoceptor vasorelaxation. Immunohistochemistry revealed greater levels of COX-1 and COX-2 staining in 54-week-old aortas compared to 8 week with expression located mainly in medial smooth muscle. An age-linked increase in COX-1 and COX-2 protein was found in cremaster arterioles of 54-week old rats (compared to 8-week) mainly in the endothelial layer. The age-related increase in COX-1 and COX-2 protein led to elevation of prostacyclin (measured as 6-keto prostaglandin F(1alpha)) and thromboxane A(2) (measured as thromboxane B(2)) in 54-week compared to 8-week-old aortas. Endothelium removal in 54-week aortas markedly reduced the 6-keto prostaglandin F(1alpha) level, thus suggesting an endothelial source for elevated prostacyclin. These findings in combination with the effects of COX inhibitors suggest that the age-related decrease in beta adrenoceptor vasorelaxation by isoprenaline is due to an age-linked increase in COX expression, which elevates production of COX-derived vasoactive prostanoids. PMID- 17404711 TI - Reevaluating the nicotine delivery kinetics hypothesis. AB - RATIONALE: The view of smoking as an addiction to nicotine implies that nicotine is an addictive drug and a primary reinforcer. However, nicotine other than in tobacco does not appear to be very rewarding for smokers. This potential anomaly to the nicotine addiction thesis is resolved by the proposition that the reward associated with smoking depends on "high-nicotine boli." According to the nicotine delivery kinetics hypothesis, smoked nicotine reaches the brain in 5-10 s in high concentrations, which provide reinforcing "hits" of nicotine to the brain. OBJECTIVES: Because of its essential role in the nicotine addiction thesis, this review set out to examine the current empirical basis of the nicotine delivery kinetics hypothesis. MATERIALS AND METHODS: We reviewed studies that bear on two questions: First, does nicotine from cigarettes reach the brain significantly faster than from other nicotine delivery devices? Second, is there a relationship between delivery kinetics and any rewarding effects of nicotine? RESULTS: There is little empirical support for the nicotine delivery kinetics hypothesis. Several studies found that arterial nicotine levels associated with smoking are much lower than predicted by the nicotine delivery kinetics thesis and not higher than with other nicotine delivery devices. More importantly, comparisons of nicotine delivery devices with varying speeds of delivery do not suggest any correlation between nicotine delivery profile and subjective reward. CONCLUSIONS: This review indicates that the wide endorsement of the nicotine delivery kinetics hypothesis is unjustified. Critical research is required to resolve the anomalies within the nicotine addiction theory of smoking. PMID- 17404713 TI - Fast ultrasound-assisted extraction of copper, iron, manganese and zinc from human hair samples prior to flow injection flame atomic absorption spectrometric detection. AB - A dynamic ultrasound-assisted extraction procedure utilizing diluted nitric acid was developed for the determination of copper, iron, manganese and zinc in human hair taken from workers in permanent contact with a polluted environment. The extraction unit of the dynamic ultrasound-assisted extraction system contains a minicolumn into which a specified amount of hair (5-50 mg) is placed. Once inserted into the continuous manifold, trace metals were extracted at 3 mL min( 1) with 3 mol L(-1) nitric acid under the action of ultrasound for 2 min for zinc and 3 min for copper, iron and manganese determination, and using an ultrasonic water-bath temperature of 70 degrees C for zinc and 80 degrees C for copper, iron and manganese determination. The system permits the direct analysis of hair and yields concentrations with relative standard deviations of <3% (n = 11). The applicability of the procedure was verified by analysing human hair samples from workers exposed to welding fumes, and its accuracy was assessed through comparison with a conventional sample dissolution procedure and the use of a certified reference material (BCR 397, human hair). PMID- 17404712 TI - Combined exposure to nicotine and ethanol throughout full gestation results in enhanced acquisition of nicotine self-administration in young adult rat offspring. AB - RATIONALE: Epidemiological evidence shows positive correlation between either maternal cigarette smoking or alcohol consumption on subsequent drug-taking behavior in offspring. However, the consequences of full gestational exposure to both drugs have not been studied experimentally despite concurrent use frequently reported among women of childbearing age. Such comorbid gestational drug exposure may increase susceptibility to acquiring cigarette smoking (i.e., nicotine self administration), a major gateway drug. OBJECTIVES: We developed a noninvasive rat model for exposure to both nicotine (2-6 mg kg(-1) day(-1)) and EtOH (4 g/kg gavage) that continued throughout pregnancy and postnatal (P) days 2-12, the rodent equivalent of the human third trimester, a critical brain developmental period. Offspring with this full gestational exposure to both drugs (Nic+EtOH) were compared to controls: nicotine alone, EtOH alone, pair-fed (comparable nutrition and handling), and ad libitum chow-fed. At P60-90, offspring had unlimited chronic access to acquire i.v. nicotine self-administration. RESULTS: There were no differences in gender ratio, stillbirths, birth weights, righting reflex, eye opening age, or weight gain. However, Nic+EtOH offspring of both genders acquired nicotine self-administration (15 or 30 microg kg(-1) injection( 1)) more rapidly, at a higher percentage, and at a higher level than offspring in the other cohorts. CONCLUSION: Full gestational Nic+EtOH exposure produced no overt alterations in standard postnatal measures but resulted in an enhanced acquisition of nicotine self-administration in young adult offspring. PMID- 17404714 TI - Porous imprinted polymer membranes prepared by phase separation in compressed liquid CO2. AB - Phase separation of poly(acrylonitrile-co-methacrylic acid) in compressed liquid CO(2) resulted in formation of a porous imprinted membrane which preferentially adsorbed uracil (URA). The cross-section of the membrane was observed by SEM, which revealed its porous structure. The mechanical strength of the membrane indicated formation of a rigid matrix with high tensile strength (4.4 N mm(-2)). The imprinted membranes bound highly selectively to URA (12.8 micromol g(-1)) but binding to dimethyluracil (DMURA), thymine, and cytosine was less (0.7, 0.8, and 0.9 micromol g(-1), respectively). When DMURA was similarly used to prepare an imprinted membrane in liquid CO(2) there was less binding of DMURA to the imprinted membrane obtained. The URA-imprinted membranes were evaluated by IR spectroscopy before and after URA extraction. The results indicated that hydrogen bonding was the mechanism of binding of URA to the imprinted membrane. Competitive binding studies were performed with binary mixtures of URA and its analogues. The URA-imprinted membrane enabled good separation of URA from cytosine, DMURA, and thymine, with separation factors of 3.0, 3.8, and 2.5, respectively. It was confirmed that the compressed liquid CO(2) contributed to efficient formation of template substrate sites in the URA-imprinted membrane. PMID- 17404715 TI - Raman spectroscopy of natron: shedding light on ancient Egyptian mummification. AB - The mummification ritual in ancient Egypt involved the evisceration of the corpse and its desiccation using natron, a naturally occurring evaporitic mineral deposit from the Wadi Natrun, Egypt. The deposit typically contains sodium carbonate, sodium bicarbonate and impurities of chloride and sulfate as its major elemental components. It is believed that the function of the natron was to rapidly remove the water from the cadaver to prevent microbial attack associated with subsequent biological tissue degradation and putrefaction. Several specimens of natron that were recently collected from the Wadi Natrun contained coloured zones interspersed with the mineral matrix that are superficially reminiscent of extremophilic cyanobacterial colonisation found elsewhere in hot and cold deserts. Raman spectroscopy of these specimens using visible and near-infrared laser excitation has revealed not only the mineral composition of the natron, but also evidence for the presence of cyanobacterial colonies in several coloured zones observed in the mineral matrix. Key Raman biosignatures of carotenoids, scytonemin and chlorophyll have been identified. PMID- 17404716 TI - Bioanalytical procedures for determination of drugs of abuse in oral fluid. AB - Recent advances in analytical techniques have enabled the detection of drugs and drug metabolites in oral fluid specimens. Although GC-MS is still commonly used in practice, many laboratories have developed and successfully validated methods for LC-MS(-MS) that can detect a large number of compounds in the limited sample volume available. In addition, several enzyme immunoassays have been commercialized for the detection of drugs of abuse in oral fluid samples, enabling the fast screening and selection of presumably positive samples. A number of concerns are discussed, such as the variability in the volume of sample collected and its implications in terms of quantitative measurements, and the drug recoveries of the many different specimen collection systems on the market. Additional considerations that also receive attention are the importance of providing complete validation data with respect to analyte stability, matrix effect, and the choice of collection method. PMID- 17404717 TI - Analytical performance of the AtheNA MultiLyte ANA II assay in sera from lupus patients with multiple positive ANAs. AB - The purpose of this study was to evaluate the precision and accuracy of a commercial multiplexed kit for the measurement of 9 anti-nuclear antibodies (ANAs; anti-SS/A, anti-SS/B, anti-Sm, anti-RNP, anti-Jo-1, anti-Scl-70, anti dsDNA, anti-Centromere B, and anti-Histone), and to compare these results to a subset of ANAs measured by enzyme-linked immunosorbent assays (ELISA) and immunodiffusion (ID). Sera were obtained from 22 systemic lupus erythematosus (SLE) patients, twelve controls and five others (commercial source) with various autoimmune diseases. ANA results from the AtheNA MultiLyte ANA II Assay (AtheNA) were compared to ELISA results (controls) and patients (ID). The AtheNA interassay coefficients of variation (CVs, N = 39, performed in duplicate; replicated 3x) ranged from 6.2% to 16.7% (mean = 9.8%), while the intra-assay CVs ranged from 5.8% to 14.3% (mean = 10.8%). Compared to results for SLE cases and controls, the sensitivity of AtheNA ranged from 85.7% to 100% (mean = 97.1%), while diagnostic specificity ranged from 16.7% to 100% (mean = 71.6%). There was significant agreement (P values ranging from 0.0001 to 0.03) when analytes coanalyzed by AtheNA and ELISA/ID were evaluated using Cohen's kappa (kappa values ranging from 0.376 to 1.000). No false positive ANA results were observed for either the control or commercial source autoimmune disease sera. These results indicate that the AtheNA assay is a precise and accurate alternative for performing multiple ELISAs or IDs in the diagnosis of autoimmune diseases, especially when the number of sera to be tested is large, such as in clinical screening or epidemiologic studies. It also appears that the AtheNA assay identifies positive ANA specificities which are missed by ID techniques, suggesting that it may have greater analytical sensitivity for some ANAs. PMID- 17404718 TI - Identification of NR1I2 genetic variation using resequencing. AB - OBJECTIVE: The nuclear receptor NR1I2 (also called PXR or SXR) is primarily expressed in mouse and human liver and intestines. Direct activation of NR1I2 occurs in response to a range of xenobiotics, which causes the formation of a heterodimer with the RXR receptor. This heterodimer binds to the nuclear receptor response elements of downstream genes such as ABCB1, CYP2C, and CYP3A. This study determined the extent of NR1I2 variation in three world populations. METHODS: Variation in NR1I2 was identified by pooled resequencing in African, Asian, and European populations. Validation was performed in European and African populations using PCR and Pyrosequencing technology. RNA expression of NR1I2, ABCB1 and CYP3A4 was assessed using real-time PCR. RESULTS: Of 36 single nucleotide polymorphisms (SNPs) identified, 24 were in the untranslated region, 8 were intronic, and 4 exonic. Thirty-six percent were unique to the African population. In comparison with previously published data, we identified 13 novel polymorphisms. The NR1I2 -566A > C polymorphism was significantly associated with ABCB1 and CYP3A4 RNA expression in colon tumor (P = 0.04 in both cases), however, this polymorphism was not associated with NR1I2 expression. CONCLUSION: With NR1I2 playing such a large role in the regulation of genes involved in drug metabolism and transport, genetic variation contributing to altered NR1I2 function may have an important clinical impact. PMID- 17404719 TI - Adverse drug reaction reporting by nurses in Sweden. AB - AIM: To investigate whether nurses could be a useful tool for improving the reporting rate of adverse drug reactions (ADRs). Furthermore, we wanted to study how physicians working at the study departments would respond to nurses as reporters of ADRs and if the reporting from the nurses affected the reporting rate from the physicians. METHOD: Three departments of internal medicine and one unit for orthopaedics were selected for the study. Nurses with special drug responsibilities were invited to participate. At the start of the study period, the nurses received an introduction with background, objective, method and other practical issues concerning the study. After this, an education programme about ADR reporting, definitions, and ADR classification according to mechanism and organ system was given. To study their knowledge about and attitude towards ADRs, a questionnaire was handed out to the nurses. A questionnaire was also handed out to all physicians at the participating departments in order to investigate their attitude towards nurses as reporters of ADRs. RESULTS: Fifty-four nurses participated in the study. During the study period, a total number of 23 reports with 39 ADRs were sent to the regional centres by the nurses. Seventeen (74%) of the reports were assessed as serious. Eight of the 39 ADRs were unlabelled and all reports were considered appropriate. The reporting rate from the physicians during the study period was similar to the previous year, indicating that the nurses contributed with additional reports. At the end of the study, the nurses thought that they had enough knowledge to report ADRs. Sixty-eight percent of the physicians did not object to nurses being included as reporters of suspected ADRs. CONCLUSION: Adverse drug reaction reporting by nurses could improve the overall safety of drugs. PMID- 17404720 TI - Modelling the influence of MDR1 polymorphism on digoxin pharmacokinetic parameters. AB - OBJECTIVES: Digoxin is a well-known probe for the activity of P-glycoprotein. The objective of this work was to apply different methods for covariate selection in non-linear mixed-effect models to study the relationship between the pharmacokinetic parameters of digoxin and the genotype for two major exons located on the multi-drug-resistance 1 (MDR1) gene coding for P-glycoprotein. METHODS: Thirty-two healthy volunteers were recruited in three pharmacokinetic drug interaction studies. The data after a single oral administration of digoxin alone were pooled. All subjects were genotyped for the MDR1 C3435T and G2677T/A genotypes. The concentration-time profile of digoxin was established using 12-16 blood samples taken between 15 min and 72 h after administration. We modelled the pharmacokinetics of digoxin using non-linear mixed-effect models. Parameter estimation was performed using the stochastic approximation EM method (SAEM). We used three methods to select the covariate model: selection from a full model using Wald tests, forward inclusion using the log-likelihood ratio test and model selection using the Bayesian Information Criterion. RESULTS: The three covariate inclusion methods led to the same final model. Carriers of two T alleles for the C3435T polymorphism in exon 26 of MDR1 had a lower apparent volume of distribution than carriers of a C allele. The only other covariate effect was a shorter absorption time-lag in women. CONCLUSION: The apparent volume of distribution of digoxin is lower in TT subjects, probably reflecting differences in bioavailability. Non-linear mixed-effect models can be useful for detecting the influence of covariates on pharmacokinetic parameters. PMID- 17404721 TI - Lateralization of functional magnetic resonance imaging (fMRI) activation in the auditory pathway of patients with lateralized tinnitus. AB - INTRODUCTION: Tinnitus is hypothesized to be an auditory phantom phenomenon resulting from spontaneous neuronal activity somewhere along the auditory pathway. We performed fMRI of the entire auditory pathway, including the inferior colliculus (IC), the medial geniculate body (MGB) and the auditory cortex (AC), in 42 patients with tinnitus and 10 healthy volunteers to assess lateralization of fMRI activation. METHODS: Subjects were scanned on a 3T MRI scanner. A T2* weighted EPI silent gap sequence was used during the stimulation paradigm, which consisted of a blocked design of 12 epochs in which music presented binaurally through headphones, which was switched on and off for periods of 50 s. Using SPM2 software, single subject and group statistical parametric maps were calculated. Lateralization of activation was assessed qualitatively and quantitatively. RESULTS: Tinnitus was lateralized in 35 patients (83%, 13 right-sided and 22 left sided). Significant signal change (P(corrected) < 0.05) was found bilaterally in the primary and secondary AC, the IC and the MGB. Signal change was symmetrical in patients with bilateral tinnitus. In patients with lateralized tinnitus, fMRI activation was lateralized towards the side of perceived tinnitus in the primary AC and IC in patients with right-sided tinnitus, and in the MGB in patients with left-sided tinnitus. In healthy volunteers, activation in the primary AC was left lateralized. CONCLUSION: Our paradigm adequately visualized the auditory pathways in tinnitus patients. In lateralized tinnitus fMRI activation was also lateralized, supporting the hypothesis that tinnitus is an auditory phantom phenomenon. PMID- 17404722 TI - Right circumflex retro-oesophageal aortic arch with coarctation of a high positioned right arch. AB - We present a rare case of right circumflex retro-oesophageal aortic arch with coarctation of a high-positioned right arch. A 7-month-old boy presented with a cardiac murmur. Cardiac situs was normal and there was no evidence of an intracardiac shunt or patent ductus arteriosus. MR aortography revealed a right aortic arch that was high-positioned, tortuous and narrowed. This right aortic arch crossed the midline behind the oesophagus and continued as a left-sided descending aorta. The left common carotid and subclavian arteries arose from a large branching vascular structure that derived from the top of the left-sided descending aorta. The right common carotid artery arose from the ascending aorta. The proximal portion of the right common carotid artery showed very severe stenosis and poststenotic dilatation. The right subclavian artery originated distal to the narrowed and tortuous segment of the aortic arch. PMID- 17404723 TI - MR imaging appearance of insulinoma in an infant. AB - Insulinoma is a very rare pancreatic neoplasm of childhood. Early recognition of insulinoma is important to ensure proper surgical treatment and prevent serious adverse neurological consequences. We describe here a case of insulinoma in an infant who presented with seizures and abnormal behaviour. Random blood glucose was found to be abnormally low (40 mg/dl). MRI showed a well-encapsulated mass (hypointense on T1-weighted and hyperintense on T2-weighted images) at the junction of the head and body of the pancreas. The tumour was successfully enucleated. Histopathology confirmed the diagnosis of insulinoma. PMID- 17404724 TI - Percutaneous thrombin embolization of a post-traumatic superficial temporal artery pseudoaneurysm. AB - Pseudoaneurysm of the superficial temporal artery is rare in children and is frequently a result of direct facial trauma. Conventional management has included surgical ligation and resection and more recently endovascular embolization to prevent complications including local pain, headache, facial disfigurement and bleeding. We report a unique case of a US-guided percutaneous thrombin injection for the treatment of a post-traumatic superficial temporal artery pseudoaneurysm in a child. PMID- 17404725 TI - Segmental omental infarction in childhood: an unusual case of left-sided location with extension into the pelvis. AB - Segmental omental infarction (OI) is a rare cause of abdominal pain in children. It generally occurs in the right lower and upper quadrants of the abdomen and only a few cases of other locations have been described in adults. We report a unique paediatric case of OI with an unusual left-sided location extending into the pelvis in a 6-year-old non-obese girl. The diagnosis was suspected on US and CT based on imaging patterns previously described in adults and children. Laparoscopic resection should be balanced with nonsurgical management. PMID- 17404726 TI - Protein engineering: opportunities and challenges. AB - The extraordinary properties of natural proteins demonstrate that life-like protein engineering is both achievable and valuable. Rapid progress and impressive results have been made towards this goal using rational design and random techniques or a combination of both. However, we still do not have a general theory on how to specify a structure that is suited to a target function nor can we specify a sequence that folds to a target structure. There is also overreliance on the Darwinian blind search to obtain practical results. In the long run, random methods cannot replace insight in constructing life-like proteins. For the near future, however, in enzyme development, we need to rely on a combination of both. PMID- 17404727 TI - Stable and continuous long-term enzymatic reaction using an enzyme-nanofiber composite. AB - This study shows the preparation and application of enzyme-nanofiber composites for long-term stable operation. The enzyme-nanofiber composite was prepared by coating an enzyme aggregate, the esterase from Rhizopus oryzae, on the surface of the nanofibers. After immobilization on the nanofiber, the apparent K ( m ) for the immobilized esterase was 1.48-fold higher than that of the free esterase, with values of 0.98 and 1.35 mM for the free and immobilized enzymes, respectively. It was found that enzyme-nanofiber was very stable, even when the fibers were shaken in glass vials, preserving 80% of the initial activity for 100 days. In addition, the enzyme-nanofiber composite was used repeatedly in 30 cycles of substrate hydrolysis and still remained active. Consequently, the esterase-nanofiber composite was employed within a continuous reactor system to evaluate its use in a long-term and stable continuous substrate hydrolysis reaction. It was found that the production of p-nitrophenol was stable for at least 400 h. This study demonstrates that the enzyme-nanofiber composite can be used in both repeated-batch mode and a continuous mode for a long-term stable operation. PMID- 17404728 TI - Metallopeptidase, neurolysin, as a novel molecular tool for analysis of properties of cancer-producing matrix metalloproteinases-2 and -9. AB - To compare the substrate preferences of rat brain neurolysin and cancer-producing matrix metalloproteinases (MMPs), which have the same architecture in their catalytic domains, the cleavage activity of neurolysin toward MMP-specific fluorescence-quenching peptides was quantitatively measured. The results show that neurolysin effectively cleaved MOCAc [(7-methoxy coumarin-4-yl) acetyl] RPKPYANvaWMK(Dnp[2,4-dinitrophenyl])-NH(2), a specific substrate of MMP-2 and MMP 9, but hardly cleaved MOCAc-RPKPVENvaWRK(Dnp)-NH(2), a specific substrate of MMP 3, suggesting that neurolysin has a similar substrate preference to MMP-2 and MMP 9. A structural comparison between neurolysin and MMP-9 showed the similar key amino acid residues for substrate recognition. The possible application of neurolysin displayed on the yeast cell surface, as a safe protein alternative to MMP-2 and MMP-9 which induce cancer cell growth, invasion, and metastasis, to analysis of properties of the MMPs, including the screening of inhibitors and analysis of inhibition mechanism etc., are also discussed. PMID- 17404729 TI - Characterization of a chitinolytic enzyme from Serratia sp. KCK isolated from kimchi juice. AB - The novel chitinolytic bacterium Serratia sp. KCK, which was isolated from kimchi juice, produced chitinase A. The gene coding for the chitinolytic enzyme was cloned on the basis of sequencing of internal peptides, homology search, and design of degenerated primers. The cloned open reading frame of chiA encodes for deduced polypeptide of 563 amino acid residues with a calculated molecular mass of 61 kDa and appears to correspond to a molecular mass of about 57 kDa, which excluded the signal sequence. The deduced amino acid sequence showed high similarity to those of bacterial chitinases classified as family 18 of glycosyl hydrolases. The chitinase A is an exochitinase and exhibits a greater pH range (5.0-10.0), thermostability with a temperature optimum of 40 degrees C, and substrate range other than Serratia chitinases thus far described. These results suggested that Serratia sp. KCK chitinase A can be used for biotechnological applications with good potential. PMID- 17404730 TI - Identification of promoters of two dehydrogenase genes in Ketogulonicigenium vulgare DSM 4025 and their strength comparison in K. vulgare and Escherichia coli. AB - Promoters of sorbose dehydrogenase gene sdh and sorbosone dehydrogenase gene sndh (Psdh and Psndh) in Ketogulonicigenium vulgare DSM 4025 were identified. The transcription initiation site (TIS) of Psdh was guanine 74 bp upstream of the start codon of sdh and the TIS of Psndh was adenine 113 bp upstream of the first codon of sndh. Comparing Psdh and Psndh, consensus sequences were found, which were TAVCVT (V=A, C or G) and THGAHC (H=A, C or T) for their putative -10 and -35 regions, respectively, and the spans between the 2 regions were 17 bp. Psdh and Psndh promoters may be constitutive in K. vulgare DSM 4025 when cultured in HJ medium. Semiquantitative RT-PCR analysis showed that the Psdh promoter was about 2.5 times stronger than Psndh in strength in K. vulgare DSM 4025. In Escherichia coli, Psdh and Psndh demonstrated strong activity with the former about two times stronger than the latter. DCIP decoloration method and reporter plasmids pSDH or pSNDH may be applied to discover promoters of genes in E. coli and to determine their strength in one step. PMID- 17404731 TI - The utility and precision of analogue and digital preoperative planning for total hip arthroplasty. AB - We prospectively compared the utility and precision of preoperative templating performed in printed films (analogue) with that performed on digital radiographs (digital) in 69 patients undergoing primary total hip replacement. Five patients were excluded when misplacement of the marker resulted in a magnification error greater that 10%; in the remaining patients (64 hips), the cup size was within +/ one size in 62 (97%) of the analogue and 52 (81%) of the digital (p=0.01) plans. The stem size was within +/- one size in 63 (98%) of the analogue and 60 (94%) of the digital (p=0.39) plans. The distance from the proximal corner of the lesser trochanter to the center of the prosthetic head (LTCD) in the analogue differed by 5 mm or more from the digital plan in nine cases (14%). Analogue preoperative planning yielded more predictable results than digital planning, particularly in terms of acetabular component size and LTCD that dictates limb lengthening shortening. The sources of error were not clearly explained by variations in magnification. Inconsistent positioning of the magnification marker may jeopardise the safe implementation of digital templating. PMID- 17404732 TI - Percutaneous release of trigger thumb: do we really need steroid? PMID- 17404733 TI - A pravastatin dose-escalation study in systemic lupus erythematosus. AB - Statin medications have been suggested for widespread use in patients with systemic lupus erythematosus (SLE). We studied the dose effectiveness and tolerability of pravastatin in SLE. We compared 41 SLE subjects in a two-month open-label dose-titration study of pravastatin to 22 SLE controls. Lipids, ALT, CPK, CRP, adverse effects were assessed. Linear mixed models assessed changes in lipids and CRP, comparing pravastatin subjects to controls. After 1 month of pravastatin 10 mg a day, total cholesterol decreased by 16% (+/-12.1%) and LDL by 24% (+/-17%), compared with 1.8% (+/-7.5%) and 2.6% (+/-8.6%) decreases in controls (P < 0.001). CRP did not decline. Glucocorticoids appeared to decrease pravastatin effectiveness. Serum CPK increased in one subject. Pravastatin reduced LDL and total cholesterol levels approximately the same degree observed in normal individuals, but the effect appeared blunted in those on modest doses of glucocorticoids and those with higher BMI. PMID- 17404734 TI - Transcription regulatory polymorphism -43T>C in the 5'-flanking region of SLC19A1 gene could affect rheumatoid arthritis patient response to methotrexate therapy. AB - The reduced folate carrier (RFC) protein (SLC19A1-gene) has central role in the uptake and intracellular accumulation of folates. In this respect, we investigate whether SLC19A1 genetic variations could affect rheumatoid arthritis (RA) patient response to antifolate treatment. One hundred six unrelated RA patients were enrolled in this study. Polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) was used as the screening method for genetic variants. Unusual SSCP patterns were characterized by direct sequencing of the PCR products and subsequently restriction assays were established. Western blot analysis of RFC protein was performed in respect of the identified SLC19A1 genotypes. Patient response to methotrexate (MTX) was evaluated using disease activity for 28 joint indices score, American College of Rheumatology 20% and 50% scores. No mutation was found in the SLC19A1 gene, but three polymorphic variants: the -43T>C in the 5'-flanking sequence to the ATG-transcription start site; and the 80G>A (R27H) and 696C>T (P232P) in the coding gene sequence. The wild type alleles of the three polymorphisms were in strict linkage disequilibrium. Western blot analysis revealed that the non-wild type allele of polymorphism -43T>C is associated with low RFC protein expression levels. Furthermore, the genotypic analysis of the functional polymorphic variant -43T>C revealed to be insufficient to predict patient response to MTX therapy. According to recent literature, several transport systems account for folate membrane transport. Additionally, in previous studies discrepancies have been reported to exist between the same genetic variants and their use in prediction of patient response to MTX therapy. Therefore, the present genotypic-phenotypic association study of a functional polymorphism revealed the need of a complex genotypic analysis in order to predict patient response to folate antagonists' therapy. PMID- 17404735 TI - Synovial fluid and peripheral blood immune complexes of patients with rheumatoid arthritis induce apoptosis in cytokine-activated chondrocytes. AB - The destruction of cartilage is an important characteristic of rheumatoid arthritis (RA). Immune complexes (IC) are usually found in high amounts in RA synovial fluids (SF) and in the superficial layers of RA cartilage. The objective of this study was to investigate if IC have a direct influence on proliferation, survival and production of nitric oxide (NO) of cytokine-activated chondrocytes. Primary bovine chondrocytes were incubated with cytokines (huIL-1alpha, bovIFN gamma, huTNF-alpha) and IC containing precipitates of peripheral blood (PB) and/or synovial fluid (SF) of 14 RA patients, 5 osteoarthritis (OA) patients and 10 healthy age and sex-matched controls. After 48 h, chondrocyte viability, proliferation, apoptosis, NO production and oxygen radical levels were measured. Staining with May-Grunwald-Giemsa after incubation with IC of RA PB and SF, showed apoptotic chondrocytes with condensation of the nuclei. The proliferation rates of cytokine-activated chondrocytes, incubated with sera and SF IC of RA patients were significantly decreased compared to chondrocytes, incubated with sera and SF IC of OA patients and compared to sera of controls. Quantitative evaluation of apoptotic cells by annexin-V/propidium iodide and TUNEL assays revealed a significant increase after incubation with sera and SF IC of RA patients, compared to control sera and OAs sera and SF. In all TUNEL positive samples, active-caspase-3-positive cells were found. There was a significant increase of chondrocyte NO production, after incubation with SF IC of RA patients, compared to OA SF. These results support the hypothesis that IC, present in serum and SF of RA patients, have a profound influence on chondrocyte growth, NO production and apoptosis, contributing to cartilage destruction in RA. PMID- 17404736 TI - Efficient procedure for grapevine embryogenic suspension establishment and plant regeneration: role of conditioned medium for cell proliferation. AB - An efficient system for the establishment and multiplication of highly prolific embryogenic cell cultures of grapevine (Vitis sp.) was developed. Using anther derived pro-embryogenic masses as starting material, cell suspensions of different grapevine cultivars (Tempranillo, Cabernet-Sauvignon) and rootstocks (Kober 125 AA, Kober 5 BB, 110 Richter) were initiated in liquid medium containing NOA (1.0 mg l(-1)) and BAP (0.25 mg l(-1)) as growth regulators. Conditioned medium was recovered and utilised for establishing new, highly totipotent cell cultures. The suspensions obtained, showed embryogenic competence resulting in somatic embryo induction and subsequent plant regeneration. In this study, a simplified establishment procedure for grapevine embryogenic cell suspension allowing the fast multiplication of embryogenic material is described. Evidence for the promoting effect of the protein fraction derived from conditioned medium, on cell proliferation was found. In bioassays, addition of ss D: -GlcY affect cell proliferation suggesting that arabinogalactan proteins are required for growth processes in grapevine cell cultures. PMID- 17404737 TI - Ultrasound imaging of non-traumatic lesions of wrist and hand tendons. AB - Non-traumatic tendon lesions are common and diverse at the wrist and hand. Improvements in high-resolution ultrasound (US) are producing increasingly high quality images of superficial structures, thus expanding the indications for this imaging modality as a tool for investigating musculoskeletal disorders. The objective of this work is to provide an update on the uses and performance of US in non-traumatic tendon disorders of the wrist and hand. The relevant anatomy is reviewed, and the normal and abnormal ultrasound scan features are described in detail, with attention not only to the tendons but also to closely related structures such as synovial sheaths and bands. The contribution of US to the evaluation of each of the most common disorders is discussed. PMID- 17404738 TI - Diffusion-weighted imaging in normal fetal brain maturation. AB - Diffusion-weighted imaging (DWI) provides information about tissue maturation not seen on conventional magnetic resonance imaging. The aim of this study is to analyze the evolution over time of the apparent diffusion coefficient (ADC) of normal fetal brain in utero. DWI was performed on 78 fetuses, ranging from 23 to 37 gestational weeks (GW). All children showed at follow-up a normal neurological evaluation. ADC values were obtained in the deep white matter (DWM) of the centrum semiovale, the frontal, parietal, occipital and temporal lobe, in the cerebellar hemisphere, the brainstem, the basal ganglia (BG) and the thalamus. Mean ADC values in supratentorial DWM areas (1.68 +/- 0.05 mm(2)/s) were higher compared with the cerebellar hemisphere (1.25 +/- 0.06 mm(2)/s) and lowest in the pons (1.11 +/- 0.05 mm(2)/s). Thalamus and BG showed intermediate values (1.25 +/ 0.04 mm(2)/s). Brainstem, cerebellar hemisphere and thalamus showed a linear negative correlation with gestational age. Supratentorial areas revealed an increase in ADC values, followed by a decrease after the 30th GW. This study provides a normative data set that allows insights in the normal fetal brain maturation in utero, which has not yet been observed in previous studies on premature babies. PMID- 17404739 TI - CT colonography: optimisation, diagnostic performance and patient acceptability of reduced-laxative regimens using barium-based faecal tagging. AB - To establish the optimum barium-based reduced-laxative tagging regimen prior to CT colonography (CTC). Ninety-five subjects underwent reduced-laxative (13 g senna/18 g magnesium citrate) CTC prior to same-day colonoscopy and were randomised to one of four tagging regimens using 20 ml 40%w/v barium sulphate: regimen A: four doses, B: three doses, C: three doses plus 220 ml 2.1% barium sulphate, or D: three doses plus 15 ml diatriazoate megluamine. Patient experience was assessed immediately after CTC and 1 week later. Two radiologists graded residual stool (1: none/scattered to 4: >50% circumference) and tagging efficacy for stool (1: untagged to 5: 100% tagged) and fluid (1: untagged, 2: layered, 3: tagged), noting the HU of tagged fluid. Preparation was good (76-94% segments graded 1), although best for regimen D (P = 0.02). Across all regimens, stool tagging quality was high (mean 3.7-4.5) and not significantly different among regimens. The HU of layered tagged fluid was higher for regimens C/D than A/B (P = 0.002). Detection of cancer (n = 2), polyps > or =6 mm (n = 21), and < or =5 mm (n = 72) was 100, 81 and 32% respectively, with only four false positives > or =6 mm. Reduced preparation was tolerated better than full endoscopic preparation by 61%. Reduced-laxative CTC with three doses of 20 ml 40% barium sulphate is as effective as more complex regimens, retaining adequate diagnostic accuracy. PMID- 17404740 TI - Double-dose 1.0-M gadobutrol versus standard-dose 0.5-M gadopentetate dimeglumine in revealing small hypervascular hepatocellular carcinomas. AB - The purpose of this study was to compare the diagnostic efficacy of double-dose 1.0-M gadobutrol with that of standard-dose 0.5-M gadopentetate dimeglumine for revealing small hypervascular hepatocellular carcinomas (HCCs). Twenty-three patients with 37 HCCs (mean size: 1.2 cm) that were diagnosed by histology (n = 13) or imaging findings (n = 10) underwent two separate 3D dynamic MRIs with 0.2 mmol/kg of gadobutrol and 0.1 mmol/kg of gadopentetate dimeglumine. Three observers interpreted both MRIs in terms of lesion detection using the alternative-free response receiver operating characteristic method and lesion-to liver contrast using matched pairs analysis. The two MRIs were also compared quantitatively by measuring the signal-to-noise ratio (SNR) of the liver and lesion as well as the lesion-liver contrast-to-noise ratio (CNR). The SNR of the liver and lesion and lesion-liver CNR with gadobutrol were better than those with gadopentetate dimeglumine (p < 0.01). However, in terms of the diagnostic accuracy (mean Az for gadobutrol: 0.878, and mean Az for gadopentate dimeglumine: 0.873), the sensitivity (92.8%), positive predictive value (92.8% vs. 93.7%) and lesion-liver contrast, the two dynamic MRIs were equivalent. Gadobutrol showed a superior degree of enhancement for hypervascular HCC than did gadopentetate dimeglumine, but the diagnostic capabilities of the two agents for revealing HCCs were equivalent. PMID- 17404741 TI - Extension patterns of nasopharyngeal carcinoma. AB - The nasopharynx represents an intersection between the nasal choanae, the oropharynx, the deep facial spaces, the skull base and the intracranial cavity. Most nasopharyngeal neoplasms are malignant tumors showing aggressive local infiltration along well-defined routes. The primary role of imaging is accurate tumor mapping and detection of possible tumor extension, especially to the skull base and the deep facial spaces. The aim of this paper is to illustrate these extension patterns of nasopharyngeal carcinomas on imaging and to show the particular implication of imaging in the correct staging of the lesion. PMID- 17404742 TI - Detection of recurrent rectal cancer with CT, MRI and PET/CT. AB - Computed tomography (CT), magnetic resonance imaging (MRI) and positron emission tomography (PET) all have the potential to directly visualize local and distant relapse of colorectal cancer (CRC). Nevertheless, the role of diagnostic imaging for routine follow-up of CRC patients remains controversial. Although MRI and PET have advantages over CT in the detection of local recurrence, until now only a few surveillance programs recommend the use of annual CT for routine follow-up. The objective of this review is to elucidate the current status of diagnostic imaging for the detection of recurrent rectal cancer based on the recent literature and our own experience. Furthermore, an insight into contemporary surveillance programs and an outlook concerning a novel technical approach to moving-table MRI at 1.5 Tesla for staging purposes are given. PMID- 17404743 TI - Multidetector-row computed tomography (MDCT) in patients with a history of previous urothelial cancer or painless macroscopic haematuria. AB - The sensitivity and specificity of MDCT for depiction and localization of urothelial carcinoma (UC) was determined retrospectively. Axial and coronal four row MDCT of the urinary tract (unenhanced, contrast-enhanced nephrographic, CT urography) was independently reviewed for UC by a radiologist (R1) and a urologist (R2), without other patient information, in 27 patients (22 male, five female; age, 72 +/- 11 years) with previous UC and/or painless macroscopic haematuria. Urinary tract segments included bladder, right and left upper, middle, and lower caliceal groups, renal pelvis, uretero-pelvic junction, upper, middle, and lower ureter. MDCT findings were corroborated by surgery, other invasive procedures, and 1-year follow-up, including MDCT, intravenous urography, and cystoscopy. Receiver-operating characteristic analysis was undertaken and the the area under the curve (AUC) calculated. Eighteen of 27 patients had evidence of UC (pTa, n = 3; pT1-pT3, n = 15; TNM 2002). Tumor was correctly located by both R1 and R2 in 17 patients (sensitivity, 94%; 95% confidence interval, 84 100%) and ruled out in seven (specificity, 78%; 95% confidence interval, 51 100%), with complete agreement. Each detected ten of 11 upper urinary tracts affected by UC. For 35 urinary tract segments with UC and 308 without, the AUC was 0.910 +/- 0.035 (R1) and 0.74 +/- 0.055 (R2), z = 2.4772, Bonferroni corrected P = 0.022. MDCT depicts urinary tracts affected by UC with high sensitivity and substantial agreement between readers with different training. PMID- 17404745 TI - Ile105Val GSTP1 polymorphism and susceptibility to colorectal carcinoma in Bulgarian population. AB - BACKGROUND AND AIMS: Etiologically, the sporadic colorectal cancer (CRC) is a complex and multifactorial disease that is linked to both exogenic and endogenic factors. Accumulating evidence indicates that susceptibility to cancers, including CRC, is mediated by genetically determined differences in the effectiveness of detoxification of potential carcinogens. A member of the glutathione-S-transferase (GST) family, GSTP1, is an important candidate for involvement in susceptibility to carcinogen-associated CRC. An A-->G transition in exon 5 of the GSTP1 gene resulting in Ile(105)Val amino acid substitution has been identified. This change leads to alteration in catalytic efficiency of variant enzyme. The aim of the current study was to evaluate the influence of Ile(105)Val GSTP1 polymorphism on susceptibility to CRC. MATERIALS AND METHODS: The GSTP1 genotyping was conducted in a case-control study of 80 ethnic Bulgarian CRC patients and 126 unaffected controls using polymerase chain reaction restriction fragment length polymorphism method. RESULTS: A statistically significant case-control difference in genotype frequencies was observed: 0.69 vs 0.54 for Ile/Ile, 0.22 vs 0.39 for Ile/Val, and 0.09 vs 0.07 for Val/Val (p = 0.049). The odds ratio (OR) for Val/Val was close to 1 (0.96, 95%CI: 0.35-2.66, p = 0.942), whereas the OR for Ile/Val was significantly lower, 0.45 (95%CI: 0.24 0.86, p = 0.016), compared to the referent Ile/Ile genotype. Although a prevalence of the GSTP1 variant allele-containing genotypes (Ile/Val or Val/Val) was found in controls than in patients (OR = 0.53, 95%CI: 0.30-0.96, p = 0.035), the allele frequencies did not show significant difference between cases and controls (p = 0.127). CONCLUSIONS: Based on the obtained protective effect of Ile/Val GSTP1 genotype, we could suggest that Ile(105)Val GSTP1 polymorphism may play some role in susceptibility to CRC. PMID- 17404746 TI - Considerations on the learning curve for laparoscopic colorectal surgery: a view from the bottom. AB - INTRODUCTION: Those who have surmounted the learning curve for laparoscopic colorectal resection state that considerable numbers of highly selected cases should comprise a department's early experience to ensure reliability of technique before routine implementation. The objective of this study was to determine how this advice may interrupt case flow. METHODS: Details on all colorectal operations performed in a single institution over a 4-year period were gleaned from a prospectively maintained database. Patient profiles were scrutinised to identify how the application of various published exclusion criteria would impact upon the theoretical completion rates of our proposed learning curve. RESULTS: In total, 317 colorectal resections were performed; 259 operations were for adenocarcinoma (including 100 rectal tumours) while 58 were for benign disease. Of those with malignancy, 25(10%) were obese, 61(24%) had previous intra-abdominal surgery, while 52(20%) were aged over 80 years and 60(23%) were ASA (3/4). Strictest exclusion criteria would halve the number of cases to be commenced laparoscopically. A specialist registrar rotating through the department would have case exposure cut from a mean of 33 to 11 in 6 months under this regimen. Prioritising benign cases in the initial experience as has been recommended by certain groups would mean that, at most, 1.2 cases would be performed every 4 weeks during the learning period. CONCLUSION: Although our caseload seems sufficient to allow the acquisition of expertise in a timely fashion, procedural flow would be markedly interrupted by stringent pre selection. A low threshold for initiating the procedure laparoscopically seems a pragmatic way of ensuring departmental confidence through familiarity. PMID- 17404747 TI - Recovery rates and functional results after repair for rectovaginal fistula in Crohn's disease: a comparison of different techniques. AB - BACKGROUND AND OBJECTIVES: Rectovaginal fistulas (RVF) in Crohn's disease continue to be a challenging problem. Several operations are often necessary to attain definitive healing of the disease process. There are no guidelines concerning optimal therapeutic approaches. Endoanal mobilization techniques such as the advancement flap technique were considered the therapy of choice for many years, but are now regarded ever more critically. We have implemented several less aggressive closure techniques that take account of the anatomy and morphology of the anorectum. The long-term results are presented in this paper. MATERIALS AND METHODS: The method used was observational analysis with a standard protocol of all patients with RVF and Crohn's disease treated surgically at a single institution. RESULTS/FINDINGS: Between January 1985 and December 2002, we treated 72 patients with low rectovaginal fistulas. The operations comprised 56 procedures performed in 37 women presenting with RVF. The patients' median age was 34.6 +/- 10 years; the follow-up period was 7.15 years (10 months-18 years). Several techniques were performed: transverse transperineal repair (n = 20), endoanal direct closure multilayer without flap (n = 15), anocutaneous flap (n = 14), and advancement mucosal or full-thickness flap (n = 7). Diverting ileostomies were created in 28 patients (76%). Recovery was achieved with the initial repair in 19 patients (51.4%). An additional 12 patients underwent repeat procedures (2-5), with an overall success rate of 27:37 (73%). The rate of recurrence was 30% during a follow-up period of 7.1 years. The rate of proctectomy was 13.5%. The success rates for each of the techniques in the above group were 70, 73, 86, and 29%, respectively. They were significantly higher with the direct closure and anocutaneous flap technique than with the advancement flap technique. However, the transperineal repair led to decreased postoperative resting pressures. In the advancement flap technique, the resting and squeezing pressure decreased significantly. The risk of developing a suture line dehiscence leading to a persisting fistula was higher in the advancement flap procedure with 43%. INTERPRETATION/CONCLUSION: Techniques with a low degree of tissue mobilization such as the direct closure and anocutaneous flap show higher success rates without significant postoperative changes in continence and manometric outcome. Impaired continence was observed only in the advancement flap group, resulting in significant changes in manometric values and recovery rates. The authors prefer to apply the direct multilayer closure technique without flap. PMID- 17404748 TI - Lichen planus remission is associated with a decrease of human herpes virus type 7 protein expression in plasmacytoid dendritic cells. AB - The cause of lichen planus is still unknown. Previously we showed human herpes virus 7 (HHV-7) DNA and proteins in lesional lichen planus skin, and significantly less in non-lesional lichen planus, psoriasis or healthy skin. Remarkably, lesional lichen planus skin was infiltrated with plasmacytoid dendritic cells. If HHV-7 is associated with lichen planus, then HHV-7 replication would reduce upon lichen planus remission. HHV-7 DNA detection was performed by nested PCR and HHV-7 protein by immunohistochemistry on lesional skin biopsies from lichen planus patients before treatment and after remission. Biopsies were obtained from lichen planus lesions before treatment (n = 18 patients) and after remission (n = 13). Before treatment 61% biopsies contained HHV-7 DNA versus 8% after remission (P = 0.01). HHV-7-protein positive cell numbers diminished significantly after remission in both dermis and epidermis. Expression of HHV-7 was mainly detected in BDCA-2 positive plasmacytoid dendritic cells rather than CD-3 positive lymphocytes. HHV-7 replicates in plasmacytoid dendritic cells in lesional lichen planus skin and diminishes after remission. This study further supports our hypothesis that HHV-7 is associated with lichen planus pathogenesis. PMID- 17404749 TI - Basic research confirms coexistence of acquired Blaschkolinear Vitiligo and acrofacial Vitiligo. AB - We report about a female patient with bilateral and unilateral blaschkolinear depigmentation on the extremities and coexistence of acrofacial vitiligo, who initially presented her first signs of depigmentation at the age of 32 years. The patient was otherwise healthy. The correct diagnosis was based on the latest up to date technology utilizing in vivo FT-Raman and Fluorescence spectroscopy, Wood's light examination of the depigmented skin and immunoreactivity of epidermal catalase expression in 3 mm punch biopsies from the linear depigmented area. The results yielded decreased catalase protein expression compared to healthy controls as well as complete absence of melanocytes. FT-Raman spectroscopy identified the presence of hydrogen peroxide (H(2)O(2)) in the mM range and Fluorescence spectroscopy demonstrated H(2)O(2)-mediated oxidation of tryptophan residues in the depigmented area. The results were in agreement with vitiligo. Repigmentation of the linear lesion was initiated after reduction/removal of epidermal H(2)O(2) with pseudocatalase PC-KUS further supporting the correct diagnosis. To the best of our knowledge this is the first case documented with vitiligo following Blaschko lines in coexistence with classical acrofacial vitiligo. This observation raises the question whether besides H(2)O(2)-mediated stress in association with genomic mosaicism could play a role in some cases with vitiligo. PMID- 17404750 TI - Replication origin plasticity, Taylor-made: inhibition vs recruitment of origins under conditions of replication stress. AB - Among his many contributions to the field of chromosome structure and dynamics, J. Herbert Taylor showed that eukaryotic cells have many more potential replication origins than they use, which they can recruit when replication forks are slowed to complete S-phase in a timely fashion. Thirty years later, his findings raise an important but largely overlooked paradox. Although new data have confirmed his results, a larger body of data has revealed that slowing replication forks activates an S-phase checkpoint cascade that inhibits initiation from unfired origins until the stress is relieved. In this paper, in celebration of Taylor's work published in Chromosoma 30 years ago, I draw attention to this paradox and offer some plausible models to explain how replication stress can both inhibit and recruit new origins. I hope that this essay will stimulate further experimentation into the basis of Taylor's original findings. PMID- 17404751 TI - A comparison of resonance tuning with positive versus negative sensory feedback. AB - We used a computational model of rhythmic movement to analyze how the connectivity of sensory feedback affects the tuning of a closed-loop neuromechanical system to the mechanical resonant frequency (omega(r)). Our model includes a Matsuoka half-center oscillator for a central pattern generator (CPG) and a linear, one-degree-of-freedom system for a mechanical component. Using both an open-loop frequency response analysis and closed-loop simulations, we compared resonance tuning with four different feedback configurations as the mechanical resonant frequency, feedback gain, and mechanical damping varied. The feedback configurations consisted of two negative and two positive feedback connectivity schemes. We found that with negative feedback, resonance tuning predominantly occurred when omega(r) was higher than the CPG's endogenous frequency (omega(CPG)). In contrast, with the two positive feedback configurations, resonance tuning only occurred if omega(r) was lower than omega(CPG). Moreover, the differences in resonance tuning between the two positive (negative) feedback configurations increased with increasing feedback gain and with decreasing mechanical damping. Our results indicate that resonance tuning can be achieved with positive feedback. Furthermore, we have shown that the feedback configuration affects the parameter space over which the endogenous frequency of the CPG or resonant frequency the mechanical dynamics dominates the frequency of a rhythmic movement. PMID- 17404752 TI - VEGF(165) and bFGF protein-based therapy in a slow release system to improve angiogenesis in a bioartificial dermal substitute in vitro and in vivo. AB - BACKGROUND: Angiogenesis can be enhanced by several growth factors, like vascular endothelial growth factor-165 (VEGF(165)) and basic fibroblast growth factor (bFGF). Delayed release of such growth factors could be provided by incorporation of growth factors in fibrin matrices. In this study, we present a slow release system for VEGF(165) and bFGF in fibrin sealant. MATERIALS AND METHODS: In vitro: Pieces of Integratrade mark matrix of 15 mm in diameter were prepared. Integratrade mark matrices were divided into four groups (A=control; B=fibrin sealant; C=fibrin sealant+growth factors; D=growth factors). In vivo: The bioartificial dermal templates were transplanted into a full-skin defect of the back of nu-nu mice. Four different groups included each six matrices at 2 and 4 weeks. RESULTS: In vitro: In groups C and D, continuous release of VEGF(165) and bFGF was eminent. The incorporation of growth factors into fibrin sealant evoked a prolonged growth factor release (p < 0.05). In vivo: A significantly higher amount of vessels was quantified in groups C and D compared to groups A and B (p < 0.001). CONCLUSIONS: A model of slow protein release by combining VEGF(165) and bFGF with fibrin sealant was produced. This model resulted in a prolonged bioavailability of growth factors in vivo for functional purposes. Fibrin and collagen can release growth factors in vivo and induce significant and faster neovascularisation in bioartificial dermal templates. PMID- 17404753 TI - Gene expression profiles are influenced by ISS, MOF, and clinical outcome in multiple injured patients: a genome-wide comparative analysis. AB - BACKGROUND: Posttraumatic immune system activation in major trauma patients is linked to systemic inflammatory response syndrome, multiple organ failure (MOF), and mortality. Recent studies suggest that genome-wide expression is altered in response to distinct clinical parameters; however, the functional allocation of theses genes remains unclear. PATIENTS AND METHODS: Thirteen patients after major trauma (Injury Severity Score < 16) were studied. Monocytes were obtained on admission (within 90 min) and at 6, 12, 24, 48, and 72 h after trauma. Complementary ribonucleic acid (RNA) targets were hybridized to Affymetrix HG U 133A microarrays. Searching for genes that are differentially expressed, the patients were dichotomously assigned depending upon survival, injury severity, and MOF. The data were analyzed by supervised analysis, clustering, and comparative pathway analysis. RESULTS: Gene expression profiles of patients with adverse outcomes (763 probe sets) mainly consist of those involved in "immunological activation" or "cellular movement," whereas the gene set associated with MOF (660) is associated with "cancer" and "cell death." Injury severity (295) leads to an overexpression of genes involved in inflammatory disease. CONCLUSION: We demonstrate for the first time a serial, sequential screening analysis of monocyte messenger RNA expression patterns after multiple injury indicating a strongly significant connection between the patients' expression profile and different clinical parameters. The latter provoke a characteristic overexpression of specific functional gene ontologies. Further studies to clarify clinical consequence of this differential gene regulation are currently anticipated. PMID- 17404754 TI - Riedel's thyroiditis: impact and strategy of a challenging surgery. AB - BACKGROUND: No surgical standard for Riedel's thyroiditis (RT) is established. Salvage surgery follows severe cervical and compressive airway symptoms or strong suspicion of malignancy. Obscured planes and multi-infiltrative extension prevent sufficient surgery with considerate complications. No alternative definitive treatment is available. In failing conservative treatment, the role of surgery in RT remains unclear. MATERIALS AND METHODS: Clinical manifestation, treatment, outcome and follow-up in a unique series of eight consecutive patients with RT are presented. RESULTS: Seven female patients and one male patient with cervical tumor growth or thyroiditis underwent four total and three sub-total thyroidectomies, respectively, one patient declined remedial surgery. Complications were one bilateral laryngeal nerve palsy and one transient hypoparathyroidism. Histology confirmed RT with perithyroidal extension and excluded malignancy in all. Symptomatic relief of cervical and airway obstruction was achieved in all. Follow-up revealed two extensive mediastinal RT recurrences 1 and 6 years after surgery. CONCLUSION: Favourable symptomatic outcome and alleviation of steroids in the majority render surgery for RT valuable when conservative treatment fails. However, more radical procedures show no advantages and recurrences are not prevented. The demanding technique in RT requires special surgical expertise and highly recommends intra-operative neuromonitoring. PMID- 17404755 TI - SLC26A7 can function as a chloride-loading mechanism in parietal cells. AB - To date three potential candidates for parietal cell basolateral Cl(-) entry have been described: the highly 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS)-sensitive Cl(-)/HCO(3)(-) exchanger AE2, the HCO(3)(-) and lowly DIDS sensitive SLC26A7 protein, and the Na(+)-2Cl(-)K(+) cotransporter (NKCC1). In this study we investigate the contribution of these pathways to secretagogue stimulated acid secretion. Individually hand-dissected rat gastric glands were microfluorimetrically monitored for Cl(-) influx and pH(i) changes. Transporter activity was determined by varying ion content and through the use of pharmacological inhibitors. Expression of SLC26A7 in rat parietal cells was shown by immunohistochemistry and Western blot. SLC26A7 was inhibited by 5-Nitro-2-(3 phenylpropyl-amino)benzoic acid (NPPB) (100 microM) in the Xenopus laevis oocyte expression system. Cl(-) influx in parietal cells was enhanced by histamine, depended partially on endogenous HCO(3)(-) synthesis and completely on extracellular Na(+). Removal and subsequent readdition of Cl(-) revealed a low and a high DIDS-sensitive HCO(3)(-) extrusion system contributing to Cl(-) uptake. At acidic pH(i), however, H(+) extrusion via the H(+),K(+)-ATPase depending on Cl(-) uptake was abolished only in the presence of 100 microM (NPPB) and at high (250 microM) DIDS concentration. There was no effect of the NKCC inhibitor bumetanide on stimulated H(+) extrusion. These results would be compatible with SLC26A7 as a Cl(-) uptake system under histamine stimulation. PMID- 17404756 TI - cyclo-Oxylipin-galactolipids in plants: occurrence and dynamics. AB - cyclo-Oxylipin-galactolipids (cGL) are mono- or digalactosyldiglycerides carrying a cyclo-oxylipin in the sn1- and/or sn2-position or esterified to the galactose moiety. These compounds were recently identified in Arabidopsis thaliana. We provide evidence that cGL are mainly, if not exclusively, part of the thylakoid and can be hydrolysed by lipolytic activities associated with photosynthesis related protein complexes in vitro. Using HPLC/ESI-mass spectrometry, cGL are shown to be restricted in occurrence to the genus Arabidopsis, they do not occur in other plants tested. A. thaliana cGL are rapidly and transiently formed upon wounding with characteristic changes in composition of the cGL-fraction. While the biological role of cGL is not understood, the genus Arabidopsis may present a model-case of chemical evolution of a novel class of regulatory molecules. PMID- 17404757 TI - Investigating a persistent coccidiosis problem on a commercial broiler-breeder farm utilising PCR-coupled capillary electrophoresis. AB - In the present study, we utilised a polymerase-chain-reaction-coupled capillary electrophoresis (CE) approach to investigate the epidemiology of Eimeria species on a broiler-breeder farm in Victoria, Australia. The Eimeria populations of two flocks vaccinated against coccidiosis were followed over an 11-week period. All seven recognised Eimeria species of chickens were detected in both flocks. One flock suffered increased morbidity and mortality in its eighth week and had consistently higher Eimeria oocyst counts, species prevalences and rates of co infections. Four Eimeria species included in the vaccine administered occurred at higher prevalences before the disease outbreak in the flock. Using the CE approach, two new, previously undescribed Eimeria genotypes were discovered in both chicken flocks, one of which dominated toward the end of the study period. The molecular approach proved versatile and capable of providing useful epidemiological data which could be used to investigate and interpret coccidiosis outbreaks. PMID- 17404758 TI - The rice OsLOL2 gene encodes a zinc finger protein involved in rice growth and disease resistance. AB - Arabidopsis LSD1-related proteins that contain LSD1-like zinc finger domains have been identified to be involved in disease resistance and programmed cell death. To investigate the potential role of LSD1-related gene in rice (Oryza sativa L.), we cloned an LSD1 ortholog, OsLOL2, from the rice cDNA plasmid library. The OsLOL2 gene is predicted to encode a polypeptide of 163 amino acids with two LSD1 like zinc finger domains with 74.5% identity to those of LSD1. Southern blot analysis indicated that OsLOL2 was a single-copy gene in the rice genome. Transgenic rice lines carrying the antisense strand of OsLOL2 with decreased expression of OsLOL2 had dwarf phenotypes, and the dwarfism could be restored by exogenous GA(3) treatment, suggesting that the dwarfism was the result of a deficiency in bioactive gibberellin (GA). In agreement with this possibility, the content of endogenous bioactive GA(1) decreased in the antisense transgenic lines. Expression of OsKS1, one of the genes encoding for GA biosynthetic enzymes, was suppressed in the antisense transgenic lines. Sense transgenic lines with increased expression of OsLOL2 were more resistant to rice bacterial blight, while antisense transgenic lines were less resistant to rice bacterial blight. The OsLOL2-GFP (green fluorescence protein) fusion protein was localized in the nucleus of cells of transgenic BY2 tobacco (Nicotiana tabacum L.). These data suggest that OsLOL2 is involved in rice growth and disease resistance. PMID- 17404759 TI - Continuing medical education: introducing four papers on vesicoureteral reflux. PMID- 17404760 TI - Neutrophil activation, protein oxidation and ceruloplasmin levels in children with Henoch-Schonlein purpura. AB - The aim of this study was to investigate the role of neutrophil activation, protein oxidation and ceruloplasmin (CLP) in the pathogenesis of Henoch-Schonlein purpura (HSP), which has not been investigated previously. Serum activities of myeloperoxidase (MPO) and arylesterase (ARYL) and levels of free thiol groups, CLP and total oxidant status (TOS) were measured in 29 children with HSP at the onset of the disease and during remission in comparison with 30 healthy subjects. Patients at active stage had significantly higher MPO activity (391+/-277 vs. 155+/-154 U/l, P<0.001), higher CLP (832+/-120 vs. 682+/-114 mg/dl, P<0.001) and TOS values (20.7+/-11.8 vs. 7.5+/-2.8 micromol H2O2/l, P<0.001) than the controls, respectively. Patients had significantly lower ARYL activity (158x10(3)+/-39x10(3) vs. 187x10(3)+/-46x10(3) U/l, P<0.001) and lower free thiol levels (234+/-48 vs. 279+/-26 micromol/l, P<0.001) than the controls, respectively. Significantly positive correlations were found between TOS and MPO (r=0.437, P=0.018) and TOS and CLP (r=0.409, P=0.028) at disease onset, whereas a negative correlation was found between MPO and thiol (r=-0.597, P=0.001) during remission. In conclusion, protein oxidation and neutrophil activation may play important roles in the pathogenesis of HSP. Further research is required to understand the potential linkage between oxidant stress and complications and to develop therapeutic strategies in HSP. PMID- 17404761 TI - Deficit irrigation promotes arbuscular colonization of fine roots by mycorrhizal fungi in grapevines (Vitis vinifera L.) in an arid climate. AB - Regulated deficit irrigation (RDI) is a common practice applied in irrigated vineyards to control canopy growth and improve fruit quality, but little is known of how imposed water deficits may alter root growth and colonization by beneficial arbuscular mycorrhizal fungi (AMF). Thus, root growth and mycorrhizal colonization were determined throughout the growing season for 3 years in own rooted, field-grown, 'Cabernet Sauvignon' grapevines exposed to three RDI treatments. Vines under standard RDI were irrigated at 60 to 70% of full-vine evapotranspiration (FVET) from 2 weeks after fruit set until harvest, a standard commercial practice. Early deficit vines were exposed to a more extreme deficit (30% FVET) during the period from 2 weeks after fruit set until the commencement of ripening (veraison), and thereafter reverted to standard RDI. Late deficit vines were under standard RDI until veraison, then exposed to a more extreme deficit (30% FVET) between veraison and harvest. The production of fine roots was reduced in both the early and late deficit treatments, but the reduction was more consistent in the early deficit vines because the additional deficit was imposed when roots were more rapidly growing. The frequency of arbuscules in fine roots was greater in both of the additional deficit treatments than in the standard RDI, a response that appeared chronic, as the higher frequency of arbuscules was observed throughout the season despite the additional deficits being applied at discrete times. It appears that grapevines compensated for a lower density of fine roots by stimulating arbuscular colonization. Irrigation did not affect yield or quality of grapes, but reduced whole-vine photosynthesis during the additional deficit periods. It appears that high-quality grapes can be produced in this region with less water than that applied under the current RDI practice because the root system of the vine may be more efficient due to greater arbuscular colonization by AMF. PMID- 17404762 TI - Physical characteristics of patients with developmental cervical spinal canal stenosis. AB - There has been only one report on the physical characteristics of patients with developmental cervical spinal canal stenosis. The objective of this consecutive clinical study was to identify the physical characteristics of patients with developmental cervical spinal canal stenosis. The subjects were 243 patients with cervical spine disease who received treatment in our department between April 2001 and March 2002. These patients were divided into two groups (the groups of patients with and without spinal canal stenosis) on the basis of their lateral cervical spine radiographs. The six items examined were height, weight, sitting height, inter inner canthal distance, upper arm length, and head circumference in each patient, and then their values were compared between the two groups. The mean inter inner canthal distance was 2.7 cm in the group of patients with spinal canal stenosis and 3.5 cm in the group of patients without spinal canal stenosis; a significant difference (P < 0.01) was observed. Regarding height, weight, sitting height, upper arm length, and head circumference, no significant difference was found while comparing the two groups. In conclusion, developmental cervical spinal canal stenosis seems to be highly likely in patients with smaller inter inner canthal distance. PMID- 17404763 TI - MRI and (18)FDG-PET in the assessment of bone marrow infiltration of the spine in cancer patients. AB - The aim of this study was to evaluate the diagnostic value of MRI and (18)FDG-PET in bone marrow infiltration of the spine due to metastases of solid tumours and lymphoma in cancer patients. In 35 cancer patients (solid tumours n = 26, lymphoma n = 9) MRI of the spine and (18)FDG-PET were reviewed and the detectability of metastases, infiltration of the spine, extent of disease, and therapeutic implications were compared. In 8/35 cases (23%) imaging technique showed concordantly no bone marrow infiltration. In 19/35 patients (54%), both MRI and (18)FDG-PET revealed bone marrow infiltration of the axial skeleton. In 12/19 patients (63%), MRI showed more extensive disease which lead to subsequent therapy. The imaging findings of MRI and (18)FDG-PET were discordant in 8/35 cases (23%). (18)FDG-PET was false positive in two patients. In six patients, (18)FDG-PET failed to detect bone metastases and bone marrow infiltration of the spine, which was detected by MRI and proven by clinical follow-up with subsequent therapy in two cases. MRI is more sensitive and specific than (18)FDG-PET detecting bone marrow metastases and infiltration of the spine and has a great impact in staging cancer patients. PMID- 17404764 TI - Accurate gas phase acidities of carboxylic acids estimated by scaling the vibrational contribution of ab initio Gibbs free energies. AB - The gas phase Gibbs free energies deltaG(T) of dissociation reaction of 14 carboxylic acids were calculated on the SCF, as well as G3 and CBS-Q levels. Corresponding accuracies were critically compared with experimental data. Since all of the results suffer from systematic errors, the procedure of scaling of thermal contribution to Gibbs free energy was applied for minimizing differences between theoretical and experimental values of deltaG(T). Two parameters were adjusted, namely the scaling of thermal contribution to Gibbs free energy of neutral and anionic forms. The presented results suggest the great effectiveness of such a procedure since for all applied basis sets within the SCF framework the achieved accuracy was below the experimental error. Besides, the proposed low cost approximation method leads to precision comparable to or even exceeding the quality offered by more sophisticated composite quantum chemistry methods. The extension of the set of training molecules up to 82 has an insignificant impact on the overall quality of deltaG(T) estimation, which suggests that a wisely chosen set of reference data may be used for the characteristics of the whole class of compounds. There is a straightforward way for the analysis of acidities/basicities of other classes of chemicals such as DNA bases, alcohols, phenols, amines, amino acids, etc. PMID- 17404765 TI - Teaching fever aphorisms: Osler revisited. PMID- 17404766 TI - Laboratory-based surveillance of patients with bacterial meningitis in Egypt (1998-2004). AB - Laboratory-based surveillance for bacterial meningitis was conducted in a network of infectious disease hospitals in Egypt to better understand the epidemiology of this infection. Healthcare and laboratory personnel were trained in basic surveillance and microbiologic processing of cerebrospinal fluid (CSF) specimens. All bacterial isolates from CSF were confirmed and tested for antimicrobial susceptibility. PCR testing was performed on a random subset of purulent, culture negative CSF specimens. Of 11,070 patients who met criteria for the case definition, 843 (8%) were culture positive (42% positive for Streptococcus pneumoniae, 20% for Haemophilus influenzae serotype b, 17% for each of Neisseria meningitidis and Mycobacterium tuberculosis, and 6% for other bacteria). Of 1,784 (46%) CSF specimens tested by PCR, 232 (13%) were positive for the first three major pathogens. Of N. meningitidis isolates, 52% belonged to serogroup A, 35% to serogroup B, and 4% to serogroup W135. S. pneumoniae isolates comprised 46 different serotypes, of which 6B, 1, 19A, 23F, and 6A were the most predominant. The overall case-fatality rate for culture-positive cases was 26% and was highest among patients with M. tuberculosis (47%). Factors significantly associated with death (p < 0.05) included admission to rural hospitals, long prodromal period, referral from other hospitals, antibiotic treatment prior to admission, and clear CSF (<100 cells/mm3). Susceptibility to ampicillin and ceftriaxone was observed in 44 and 100% of H. influenzae serotype b isolates and in 52 and 94% of S. pneumoniae isolates, respectively. This surveillance highlights the significant mortality and morbidity associated with bacterial meningitis in Egypt. Decision makers need to review current treatment guidelines and introduce appropriate vaccines for prevention and control of the disease. PMID- 17404767 TI - Explorations of anticipatory behavioral control (ABC): a report from the cognitive psychology unit of the University of Wurzburg. AB - The report comprises recent theoretical considerations, experimental research, and simulations which all aim at a clarification of anticipatory mechanisms of behavioral control. PMID- 17404768 TI - Rapid analysis of acylglycerols in low molecular weight milk fat fractions. AB - A suitable analytical method was required to facilitate development of an industrial-scale short-path distillation (SPD) process. Short-path distillation produces milk fat distillates (MFD) enriched in low molecular weight milk fat components-viz. free fatty acids, monoacylglycerols, diacylglycerols, cholesterol and low molecular weight triacylglycerols. In this case, solid-phase extraction (SPE) was considered a better alternative than thin-layer chromatography for separating polar and apolar lipid components in MFD samples due to its speed and near-complete recoveries. Solid-phase extraction of MFDs yielded two fractions, both of which are sufficiently pure for subsequent analysis by gas chromatography. This procedure provided rapid and complete chemical characterization (including mass balances) of low-molecular weight milk-fat fractions. PMID- 17404769 TI - Fatty acids in liver, muscle and gonad of three tropical rays including non methylene-interrupted dienoic fatty acids. AB - Scientific investigation of lipids in Elasmobranchs has been conducted mainly on shark species. Because rays seem to be neglected, this study was performed to examine the complete fatty acid (FA) composition with a particular interest for long-chain polyunsaturated FA (PUFA) content in different tissues of three ray species including parts usually discarded. The total FA and PUFA profiles of total lipids were determined in muscle, liver, and gonad of Rhinobatos cemiculus, Rhinoptera marginata, and Dasyatis marmorata, the most often caught ray species from the East Tropical Atlantic Ocean. Fifty FA were characterized as methyl esters and N-acyl pyrrolidides by gas chromatography/mass spectrometry, showing significant levels of 20:5n-3 (EPA) (up to 5.3%) and 22:5n-3 (DPA) (up to 7.3%), high levels of 20:4n-6 arachidonic (ARA) (4.8-8.6% of total FA) and 22:6n-3 (DHA) (up to 20.0%). The results show that muscle, liver and gonad of rays can provide high amounts of essential PUFA, specially DHA, for direct human nutrition or the food processing industry. High proportions of DHA were particularly found in all samples of R. cemiculus (11.6-20.0%), and in muscle and liver of D. marmorata (11.1-16.1%). Regarding the high amounts of (n-3) PUFA, this study shows that these rays deserve a better up-grading, including the normally discarded parts, and describes the occurrence of unusual NMID FA in all tissues studied. Five non methylene-interrupted dienoic fatty acids (NMID FA) (0-3.4%) were reported, including previously known isomers, namely 20:2 Delta7,13, 20:2 Delta7,15, 22:2 Delta7,13, 22:2 Delta7,15, and new 22:2 Delta6,14. These acids are quite unusual in fish and unprecedented in rays. The 22:2 Delta6,14 acid occurred in gonads of male specimens of R. cemiculus at 2.9%. PMID- 17404772 TI - Syllabus for postgraduate specialisation in Nuclear Medicine: 2006/07 update. PMID- 17404770 TI - Targeted functional imaging in breast cancer. PMID- 17404773 TI - Trends in laryngopharyngeal reflux: a British ENT survey. AB - There is a lot of scepticism surrounding laryngopharyngeal reflux (LPR). Symptoms such as globus pharyngeus, constant throat clearing, chronic cough, idiopathic hoarseness, catarrh and choking episodes may be reflux-related. The aim of this survey was to highlight current treatment trends in LPR. Questionnaires were emailed to 260 members of the British Academy of Otolaryngology-Head and Neck surgery (BAO-HNS). Survey recipients were asked about type, duration and dose of antireflux treatment and length of follow-up appointments, if any. Finally, they were asked about awareness of any reflux symptom and reflux sign questionnaires. Survey response rate was 60%. The vast majority of the otolaryngologists surveyed believe in laryngopharyngeal reflux (90%) and more than 50% prescribe proton pump inhibitors (PPIs). The preferred duration of treatment is 2 months (37%). Only a minority will prescribe PPIs for 6 months or more. Most otolaryngologists will give the standard GORD dose (70%) (once daily) and only a few (20%) will prescribe more aggressive and prolonged doses. The commonest symptoms for which proton pump inhibitors are prescribed are globus (73%), followed by choking episodes (66%) and chronic cough (62%). If LPR is suspected, most of the otolaryngologists will follow-up the patients (61%) and approximately one third (31%) will discharge them back to the general practitioners. Only eight-percent 8% will refer to gastroenterologists. The three commonest laryngoscopic signs that makes them suspect LPR are erythema of the arytenoids (86%) or the vocal cords (57%) and granulomas (42%). The majority of the otolaryngologists (94%) do not use popular questionnaires such as the RFS or RSI. Despite the controversy surrounding laryngopharyngeal reflux, our results suggest that the majority of the otolaryngologists surveyed believe in LPR and attempt to treat it. Interesting findings are: the duration of treatment, the doses used, the length of follow-ups or the lack of, and the fact that the majority does not request any specific diagnostic tests. "symptoms and signs" questionnaires are rarely used. PMID- 17404775 TI - An intracerebral intraparenchymatous triton tumor in a man with neurofibromatosis. AB - We report the second case of a cerebral intraparenchymatous malignant nerve sheath tumor, triton tumor variant. It is the first case associated with neurofibromatosis reported in literature thus far. The therapy of these aggressive tumors is rather disappointing as recurrence of the tumor despite complete surgical resection and even adjuvant radiotherapy and/or chemotherapy is seen in most patients. It is still unknown if survival is influenced by tumor localization, size or malignancy grade. In this case therapy also proved to be unsuccessful. PMID- 17404776 TI - Analysis of spectrum and frequencies of mutations in McArdle disease. Identification of 13 novel mutations. AB - BACKGROUND: McArdle disease, a common metabolic myopathy with autosomal recessive inheritance, is caused by a frequent R50X mutation and many rare mutations in the myophosphorylase gene. OBJECTIVES: To identify spectrum and frequencies of myophosphorylase gene mutations in a large cohort of patients with McArdle disease, to discuss diagnostic implications, and to analyse genotype-phenotype relationship. METHODS: Molecular genetic analysis of 56 index patients with muscle biopsy-proven myophosphorylase deficiency from Germany (n = 35), UK (n = 13), and several other countries (n = 8) was performed using direct sequencing. RESULTS: Allele frequency of the R50X mutation was 58%, and 71% of the patients carried this mutation at least on one allele. We detected 26 other less common mutations, 13 of which are novel: G157V, R161C, Q337R, E384K, S450L, G486D, R570W, K575E, IVS6-2A>T, IVS10+1G>A, R650X, c.1354insC, c.1155_1156delGG. There was no genotype-phenotype correlation with respect to age of onset and severity. R270X was the most frequent mutation among the less common mutations reaching an allele frequency of 5% followed by R94W and G686R representing a frequency of 4% each. CONCLUSIONS: The study further extends the genetic heterogeneity of myophosphorylase gene mutations showing no mutational hotspot and no genotype phenotype correlation. Most novel missense mutations were located in secondary structures or active sites of the enzyme. Some of the less common mutations are recurrent with different frequencies within Europe. Ethnic origin and frequency of less common mutations must be considered to establish efficient strategies in molecular genetic testing. Performing molecular testing can avoid muscle biopsy. PMID- 17404777 TI - Episodic memory impairment in patients with Alzheimer's disease is correlated with entorhinal cortex atrophy. A voxel-based morphometry study. AB - The aims of this study were to investigate the pattern of cortical atrophy and the relationships between memory performances and the brain regions in Alzheimer's Disease (AD). optimized voxel-based morphometry (VBM) was applied to the MRI brain images of 18 probable AD and 18 healthy subjects (HS). Patients performed verbal and visuo-spatial episodic and shortterm memory tests. Contrasting of AD group with HS, and anatomobehavioural correlations were carried out in order to identify regional atrophic changes and neuro-cognitive aspects in AD group. We found evidence of gray matter (GM) volume reduction in AD in the medial temporal, parietal and frontal areas bilaterally and in the left anterior thalamic nuclei. Performance on the episodic memory delayed recall tests co varied with GM volume in the left entorhinal cortex. The pattern of cortical atrophy likely reflects the heterogeneous level of dementia severity in our AD group. The anatomical region affected in the left hemisphere indicates a sufferance at multiple levels of the Polysynaptic Hippocampal Pathway, which is involved in declarative memory. Findings on the entorhinal cortex and the delayed memory scores support the role of the entorhinal cortex in episodic memory. Damage to the entorhinal cortex, deafferenting the hippocampus from neocortical inputs, interferes with episodic memory consolidation in AD patients. PMID- 17404778 TI - Secondary "hypnic headache". AB - Hypnic headache is a rare form of primary headache with attacks occurring exclusively with sleep. As it is typical of the elderly, a wide range of alternative diagnosis must be considered. We present a case report of a 54-year old woman with unsuspected secondary hypnic headache that was relieved by anti hypertensive therapy. We reviewed the literature to evaluate the usual diagnostic workup performed in hypnic headache patients. We suggest that 24 h ambulatory blood pressure monitoring may be included in the evaluation of nocturnal headache complaints especially in the elderly, in whom essential hypertension is a very frequent comorbidity. PMID- 17404779 TI - Effect of exogenous melatonin on sleep and motor dysfunction in Parkinson's disease. A randomized, double blind, placebo-controlled study. AB - Insomnia, sleep fragmentation and excessive daytime sleepiness are common in Parkinson's disease (PD) and may contribute to the reduction of cognition and alertness in those patients. Melatonin has been shown to improve sleep in several conditions. In experimental models of PD, melatonin can ameliorate motor symptoms. To evaluate the effect of melatonin on sleep and motor dysfuntion in PD, we studied 18 patients (Hoehn & Yahr I to III) from a PD clinic. Prior to treatment, motor dysfunction was assessed by UPDRS II, III and IV. Subjective sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI) and daytime somnolence by the Epworth Sleepiness Scale (ESS). Full polysomnography (PSG) was performed in all subjects. Patients were then randomized to receive melatonin (3mg) or placebo one hour before bedtime for four weeks. All measures were repeated at the end of treatment. On initial assessment, 14 patients (70%) showed poor quality sleep (PSQI > 6) and eight (40%) excessive daytime sleepiness (ESS > 10). Increased sleep latency (50%), REM sleep without atonia (66%), and reduced sleep efficiency (72%) were found on PSG. Eight patients had an apnea/ hipopnea index greater than 15 but no severe oxygen desaturation was observed. Sleep fragmentation tended to be more severe in patients on lower doses of levodopa (p = 0.07). Although melatonin significantly improved subjective quality of sleep (p = 0.03) as evaluated by the PSQI index, PSG abnormalities were not changed. Motor dysfunction was not improved by the use of melatonin. Undetected differences in motor scores and PSG findings may have been due to a small sample size and a type II error. PMID- 17404780 TI - Too hot to handle: heat stroke in multiple system atrophy. PMID- 17404781 TI - Vertebral body integrity: a review of various anatomical factors involved in the lumbar region. AB - The body of the vertebra can be affected in the majority of the conditions involving the lumbar spine. Multiple references, both books and periodicals, have been reviewed, and the anatomical factors responsible for the vertebral body integrity in the lumbar spine have been included under the following important areas, namely, morphology, development, genetics, microscopic examination using histology, structural architecture, blood supply, neuromuscular control, and biomechanics. INTRODUCTION: The anatomy provides a three-dimensional frame work to support the interaction between the physiological and pathological alterations. The body of the vertebra can be affected in a majority of acute or chronic conditions involving the lumbar spine. The etiology of these conditions is multifactorial, which has been dealt with in previous studies sporadically. This study aims to review and incorporate the important anatomical factors which can influence the integrity of vertebral bodies in the lumbar region and manifest as low back pain. METHODS: Multiple references, both books and periodicals, have been reviewed for the literature. Electronic databases, including Medline and PubMed, were used to collect the latest information. They were finally arranged in an anatomical framework for the article. An attempt has been made to cover these relevant issues in an integrated way in the article and have been structured into introduction, morphology, development, genetics, microscopic examination using histology, structural architecture, blood supply, neuromuscular control, biomechanics, and conclusion. The aforementioned anatomical aspects, some of which have received less attention in the literature, may be helpful to clinicians for restoring the mobility, stability, and load bearing capacity of the lumbar spine as well as planning better management strategies, especially for the chronic low back pain. RESULTS: In our article all the anatomical factors affecting the integrity of vertebral body, including the morphology, development, genetics, growth and ossification, blood supply, specifically in the lumbar region, have been described, which were not covered earlier. The limitations of this review is its wide dimensions; hence, there are fair scopes of missing many relevant facts, as all of them cannot be compiled in a single article. We have attempted to confine our views to different anatomical domains only, this is our second limitation. Additional studies are required to incorporate and discuss the uncovered relevant scientific details. CONCLUSIONS: The integrity of the body of the lumbar vertebra is multifactorial (Fig. 8). The vast spectrum of the anatomical domain influencing it has been summarized. The evolution of erect posture is a landmark in the morphology of human beings and the lumbar lordosis, which has also contributed to the gross design of the vertebral body, is one of the most important adaptations for axial loading and bipedal movements. The role of metamerism in the evolution of vertebrate morphology is repeated in the development of spine. The body of the vertebra is intersegmental in origin, which results in dual vascular and nerve supply, both from superior and inferior aspects of the body of the lumbar vertebrae. The vertebral body ossifies from three primary centers, one for centrum, which will form the major portion of body, and the other two for neural arches. The cartilaginous growth plate is mainly responsible for the longitudinal vertebral growth. Regional differentiation of the vertebral column, and the definite pattern of the structure of the different vertebra, is regulated by a large number of genetic factors, including the Hox genes. The vertebral body design therefore provides the requirements for optimal load transfer by maximal strength with minimal weight. Bone mineral density (BMD), bone quality, microarchitecture, and material properties are the important factors that contribute to bone strength. BMD is highly heritable; bone mineral distribution and architecture are also shown to be under strong genetic influence. All the aforementioned factors finally integrate to ensure mainly the mobility, stability, and load bearing capacity of the lumbar spine. PMID- 17404782 TI - Collapsing focal segmental glomerulosclerosis in a liver transplant recipient on alendronate. AB - We describe a case of collapsing focal segmental glomerulosclerosis and severe kidney dysfunction in a liver transplant recipient after the initiation of alendronate for osteopenia. In view of the increasing incidence of chronic kidney disease in long-term liver transplant patients, bisphosphonates need to be used with caution in these patients. The usefulness of bisphosphonates for the prevention of early bone loss after liver transplantation is increasingly reported. However, there is little information on the safety and efficacy of these drugs when used in the later stages of liver transplant, particularly in the presence of chronic kidney disease. Bisphosphonates are excreted unchanged via the kidneys after reaching the systemic circulation. Some cases of severe kidney injury, in particular collapsing focal segmental glomerulosclerosis, have been described that are associated with the use of pamidronate. Alendronate, a widely used bisphosphonate in transplant patients, has not been related to kidney toxicity. We describe a case of collapsing focal segmental glomerulosclerosis and severe kidney dysfunction in a liver transplant recipient soon after the initiation of alendronate for osteopenia. Possible pathogenetic mechanisms are discussed. In view of the increasing incidence of chronic kidney disease in long term liver transplant patients, bisphosphonate need to be used with caution in patients with a low glomerular filtration rate. PMID- 17404783 TI - Effects of an educational-behavioral joint protection program on people with moderate to severe rheumatoid arthritis: a randomized controlled trial. AB - The aim of this study was to asses the effects on pain, disability, and health status of an educational-behavioral joint protection program in a group of moderate-severe rheumatoid arthritis (RA) patients. Eighty-five subjects with RA in treatment with anti-tumor necrosis factor alpha (TNFalpha) drugs (infliximab) were enrolled into the study and randomized into either an experimental group (46, EG) or a control group (39, CG). We organized four EG meetings, which included information on pathophysiology and evolution of RA, joint protection during normal activities of daily living, suggestions on how to adapt the surrounding environment, and self-learning exercises to perform at home. Sociodemographic characteristics and degree of knowledge of the disease, measured by the Health Service Interview (HSI), were recorded at baseline. The outcome measures included the Visual Analogue Scale (VAS), the Arthritis Impact Measurement Scale 2 (AIMS2), and the Health Assessment Questionnaire (HAQ), which were administered at the beginning and end of the trial. Thirty-six patients from the EG (7 men and 29 women; mean age 54.2 years) and 34 from the CG (6 men and 28 women; mean age 52.2 years) completed the trial. No statistical differences in baseline evaluations were found between the two groups. According to the answers given on the HSI, the majority of our patients had poor knowledge of RA and its consequences. After a mean time of 8 months, the patients receiving educational training displayed a significant decrease, compared to the CG, in the VAS (p = 0.001), HAQ (p = 0.000), and physical (p =0.000), symptoms (p = 0.049), and social interaction (p = 0.045) scores on the AIMS2, but not in other items. Our study showed that 8 months after attending an educational-behavioral joint protection program, subjects with moderate-severe RA presented less pain and disability and thus an enhanced health status. This approach may efficiently complement drug therapy in these patients. PMID- 17404784 TI - Performance of the osteoporosis risk assessment tool in Moroccan men. AB - Measuring bone mineral density (BMD) is a widely accepted strategy for identifying subjects with an increased risk of fracture. However, because of limited availability of BMD technology in some communities and cost considerations, it has been proposed that BMD measurements be targeted to subjects with risk factors for osteoporosis. Osteoporosis self-assessment tool (OST) using age and weight have been developed to identify women who are more likely to have low BMD and thus undergo BMD testing. To study the performance of OST in identifying osteoporotic white men in Morocco. We analysed in an epidemiological cross-sectional study the records for 229 white Moroccan men seen at an out-patient rheumatology centre. OST was compared to bone density T scores and the ability of OST to identify men with osteoporosis (T < -2.5) was evaluated. Using an OST score < 2 to recommend dual X-ray absorptiometry (DXA) referral, sensitivity ranged from 63% at the lumbar spine to 87% at the total hip to detect BMD T scores of -2.5 and specificity from 58 to 59%. The negative predictive value was high at all skeletal sites (87-98%), demonstrating the usefulness of the OST to identify patients who have normal BMD and should not receive DXA testing. The performance of OST among men in Morocco was similar to that reported earlier for the other samples in Asian countries and the USA. The OST is an effective and efficient tool to help target high-risk men for DXA measurement. PMID- 17404785 TI - Relapsing polychondritis presenting as meningoencephalitis with valvular abnormality: a case report. AB - Relapsing polychondritis is a rare rheumatologic disorder that is characterized by recurrent inflammation of selected connective tissue sites and destruction of cartilage throughout the body. We report a case of newly diagnosed relapsing polychondritis in a 40-year-old man presenting with episcleritis, deformed "cauliflower" ears, aortic regurgitation, and aseptic meningoencephalitis. Steroid therapy was instituted with good resolution of his clinical symptoms. PMID- 17404786 TI - Environmental microbial ecology in an "omics" era: summary report of MicroEnGen II (Shanghai, June 12-15, 2006). PMID- 17404787 TI - Isolation and characterization of four gram-positive nickel-tolerant microorganisms from contaminated sediments. AB - Microbial communities from riparian sediments contaminated with high levels of Ni and U were examined for metal-tolerant microorganisms. Isolation of four aerobic Ni-tolerant, Gram-positive heterotrophic bacteria indicated selection pressure from Ni. These isolates were identified as Arthrobacter oxydans NR-1, Streptomyces galbus NR-2, Streptomyces aureofaciens NR-3, and Kitasatospora cystarginea NR-4 based on partial 16S rDNA sequences. A functional gene microarray containing gene probes for functions associated with biogeochemical cycling, metal homeostasis, and organic contaminant degradation showed little overlap among the four isolates. Fifteen of the genes were detected in all four isolates with only two of these related to metal resistance, specifically to tellurium. Each of the four isolates also displayed resistance to at least one of six antibiotics tested, with resistance to kanamycin, gentamycin, and ciprofloxacin observed in at least two of the isolates. Further characterization of S. aureofaciens NR-3 and K. cystarginea NR-4 demonstrated that both isolates expressed Ni tolerance constitutively. In addition, both were able to grow in higher concentrations of Ni at pH 6 as compared with pH 7 (42.6 and 8.5 mM Ni at pH 6 and 7, respectively). Tolerance to Cd, Co, and Zn was also examined in these two isolates; a similar pH-dependent metal tolerance was observed when grown with Co and Zn. Neither isolate was tolerant to Cd. These findings suggest that Ni is exerting a selection pressure at this site for metal-resistant actinomycetes. PMID- 17404788 TI - Seasonal variations in planktonic community structure and production in an Atlantic coastal pond: the importance of nanoflagellates. AB - The structure and summertime production of planktonic communities and the role of nondiatom planktonic cells were studied in coastal ponds, which are areas traditionally used for fattening and greening table-sized oysters. The abundance and biomass of nano-microplanktonic protists were determined at weekly intervals between February 1998 and February 1999 in a coastal pond without oysters in the French Atlantic coast near La Rochelle. The production of these microbiotas was determined in the summer period. The structure of plankton communities revealed the following observations: (1) microphytoplanktonic cells were mostly diatoms and dinoflagellates, (2) microzooplanktonic cells were mainly ciliates, and (3) nanoplanktonic cells were represented by pigmented (80-90% of the nanoplankton biomass) and colorless nanoflagellates. Diatoms were dominated by Naviculiineae. Dinoflagellates were dominated by Peridiniales. Oligotrichida were predominant in the ciliate community. Protist biomass levels were nine times higher from April to August (summer period 1033 microg C L(-1)) than from September to March (winter period 114 microg C L(-1)). Whatever the season, nanoflagellates were dominant in the water column (66 and 53% of the entire protist biomass in the summer and winter periods, respectively). Nanoflagellates represented the highest production of nano-microplanktonic communities (76% of carbon protist production) in the coastal pond in summer and showed the shortest generation time (7.1 h). Dinoflagellates came after nanoflagellates in production (19.5% of carbon protist production). Diatoms represented only a supplementary carbon resource available for higher trophic levels, whereas, until now, they were considered as the principal food of oysters in coastal ponds. Ciliates were a small source of carbon, but their growth rate was high. We suggest, first, that nanoflagellates represented the primary resource available in the pond and could constitute an important food resource for higher trophic levels, such as oysters, farmed in this type of pond. Overall, the system appeared to be more autotrophic than heterotrophic. Because inorganic nutrients are quickly exhausted in a semiclosed pond, pigmented flagellates dominated the carbon biomass, production and biomass of bacteria were high (thus, the microbial food web appeared to be active in this pond), and mixotrophy seemed to be an important trophic mode there. PMID- 17404789 TI - PET-CT vs. CT alone in ovarian cancer recurrence. AB - BACKGROUND: To compare fusion, positron emission tomography-computed tomography (PET-CT) with CT alone in detecting ovarian carcinoma recurrence. METHODS: Fifty one consecutive patients underwent 53 restaging PET-CT scans with a concurrent diagnostic quality CT scan. Two body imaging radiologists independently assessed the CT's; each then teamed with a nuclear medicine specialist to review the PET CT's. Two teams conferred for consensus on the presence of disease in the chest, abdomen, and body overall with CT alone and with PET-CT, using a six-point reader confidence metric to determine accuracy and receiver operating characteristic (ROC) curves. Reader agreement was compared using kappa. Recurrence was determined by two gynecologic oncologists reviewing clinical records from time of presentation to at least 13 months (mean 22.7) after imaging. RESULTS: Recurrence was based on histopathology in 17% (9/53). Seventy-two percent (38/53) cases had recurrence, with two cases showing isolated chest recurrence. PET-CT accuracy exceeded CT for body 92% (49/53) vs. 83% (44/53), chest 96% (51/53) vs. 89% (47/53), and abdomen 91% (48/53) vs. 79% (42/53). ROC curves for PET-CT dominated that for CT alone; this difference was statistically significant for abdomen and for body overall (P < 0.01). Interobserver agreement was better for PET-CT than for CT alone. CONCLUSIONS: PET-CT demonstrates greater accuracy and less interobserver variability than CT alone. PMID- 17404790 TI - PEG "Rescue": a practical NOTES technique. AB - Dislodged percutaneous endoscopic gastrostomy (PEG) tubes occur commonly and may require urgent surgical intervention in a susceptible patient population. Natural orifice translumenal endoscopic surgery (NOTES) may facilitate PEG rescue and avoid the morbidity associated with contemporary surgical techniques. We report a case of a dislodged PEG tube in the early post-operative period with evidence of incomplete gastrocutaneous tract formation and intra-abdominal leakage. Bedside transgastric NOTES exploration facilitated peritoneoscopy, evacuation of intra abdominal fluid, and re-establishment of the PEG tube through the original gastrotomy tract. Tube feeds were resumed and postoperative contrast fluoroscopy demonstrated no intra-abdominal leakage from the replaced PEG tube. No postoperative complications related to the NOTES procedure were noted at 30 days of follow-up. PEG rescue represents a unique, practical, and empowering application of the burgeoning experience of NOTES. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi: 10.1007/s464-007-9361-2) contains supplementary material, which is available to authorized users. PMID- 17404791 TI - Foreign material erosion after laparoscopic Roux-en-Y gastric bypass: findings and treatment. AB - BACKGROUND: Patients after laparoscopic Roux-en-Y gastric bypass (LRYGB) can have nonspecific, upper gastrointestinal (UGI) complaints. During postoperative endoscopy, we have noted the erosion of nondissolvable material, such as Peri Strips or silk sutures, into the gastric pouch. This study reports the incidence and presentation of foreign material erosion into the gastric pouch after a LRYGB and the outcome after therapeutic endoscopy. METHODS: From a prospective LRYGB database, postoperative endoscopies from February 2002 to June 2005 that found foreign material in the gastric pouch were reviewed. Presenting symptoms, time until endoscopy, and patient outcomes were evaluated. A therapeutic endoscopy was performed by using endo-shears and biopsy forceps to remove the foreign material. RESULTS: A total of 23 patients underwent 29 therapeutic endoscopies. From February 2002 to November 2004, 208 LRYGB were performed using silk suture for the outer layer of the gastrojejunostomy, and 21 patients (10%) had silk suture found on endoscopy. Peri-Strips were used from February 2002 to December 2003 (n = 153) and 6 patients (4%) had erosions. Since converting to Vicryl suture and Seamguard, we have had 173 and 228 patients, respectively, without pouch erosion. The most common presenting symptom was abdominal pain (n = 15, 65%). Other symptoms included nausea (n = 13, 57%), vomiting (n = 12, 52%), dysphagia (n = 5, 22%), and melena (n = 3, 13%). Foreign material was found by a mean of 34 weeks. Of 29 therapeutic endoscopies, 20 resulted in resolution of symptoms (69%), 4 led to improvements (14%), and 5 had no effect (17%). There were no endoscopic complications and no anastomotic leaks. CONCLUSIONS: Nonabsorbable material used during a LRYGB can migrate into the gastric pouch and cause UGI complaints. A therapeutic endoscopy will resolve most UGI symptoms. Using absorbable material in the creation of the gastric pouch and the gastrojejunostomy will avoid foreign material erosion. PMID- 17404792 TI - Successful new method of extracorporeal percutaneous endoscopic gastrostomy (E PEG). AB - BACKGROUND: Although percutaneous endoscopic gastrostomy (PEG) has become popular for patients with swallowing disorders as a nutrition support or a decompressant of gastrointestine, perioperative complications associated with PEG have not decreased, especially peristomal infections. To reduce peristomal infections, we designed a new method of gastrostomy by extracorporeal approach under endoscopic observation, named as extra-corporeal PEG (E-PEG). METHODS: Experimental studies for E-PEG were performed repeatedly using pigs under general anesthesia to confirm the safety of its procedure for human use. After approval of institutional ethics review board in our university, thirty patients with prior consent participated in this study. The operation time, the incidence rate of complications and the hospital stay were compared between E-PEG and ordinary pull method PEG groups. RESULTS: Two patients (6.7%) in E-PEG group had postoperative complications, i.e., aspiration pneumonia and surgical site infection. The operation time of E-PEG group was 5-16 (mean +/- SD: 10.3 +/- 2.96) min as compared to 14-37 (mean +/- SD: 26.9 +/- 8.39) min with pull-method PEG. The postoperative hospital day of E-PEG was within two days except for the two complicated cases. Significance differences of operation time, complication rate and postoperative hospital stay between those groups observed statistically. CONCLUSIONS: These results indicate that E-PEG was safe, tolerable and speedy when compared ordinary pull-method PEG. PMID- 17404793 TI - Predictors of outcome after obesity surgery in New York state from 1991 to 2003. AB - BACKGROUND: This study aimed to examine hospital volume in New York state as a predictor of complications over 13 years. METHODS: Data from the New York state Statewide Planning and Research Cooperative Systems database were analyzed by diagnosis-related group, patient demographics, and outcomes. The institution data were analyzed on the basis of volume and logistic regression using chi-square analysis for predicting morbidity and mortality. RESULTS: From 1991 to 2003, 24,534 patients underwent bariatric surgery at 79 institutions. Case volume increased 36-fold in high-volume centers (HVCs), whereas the overall number of institutions doubled. The average mortality rate was 1.1% in low-volume centers (LVCs), 0.4% in medium-volume centers (MVCs), and 0.2% in HVCs. Whereas 12.4% of the LVC patients had postoperative anastomotic complications (PACs), only 9.7% had PACs in MVCs and only 7.3% had PACs in HVCs (p < 0.05). Furthermore, only 0.7% of the patients required postoperative reintubation (PRI) after surgery at HVCs, as compared with 1.5% at MVCs and 1.8% at LVCs (p < 0.05). Finally, 1.8% of the patients required blood transfusion of packed red blood cells at HVCs, whereas 2.4% required transfusion at MVCs and LVCs. CONCLUSIONS: Experience correlates with fewer postoperative complications, including death, in bariatric centers in New York state. PMID- 17404794 TI - Comparing T2 and T2-T3 ablation in thoracoscopic sympathectomy for palmar hyperhidrosis: a randomized control trial. AB - BACKGROUND: Thoracoscopic sympathectomy is a useful therapeutic option for palmar hyperhidrosis. Surgeons differ in the level of the sympathetic chain ablated. This study aimed to compare the blockade of the T2 with levels T2 and T3 to verify the effectiveness of different ablation levels in relieving hyperhidrosis symptoms. METHODS: For patients undergoing bilateral thoracoscopic sympathectomy for palmar hyperhidrosis, T2-T3 ablation is performed bilaterally. In our series, 25 consecutive patients were blindly randomized to undergo unilateral T2 and T3 ablation followed by contralateral ablation of level T2 only. The patients were followed up and analyzed for comparison of symptoms bilaterally, compensatory hyperhidrosis, and levels of satisfaction postoperatively. RESULTS: The study group consisted of 25 patients with a male:female ratio of 3:2 and a mean age of 32 years (range, 19-50 years). The mean operative time was 35 min. The patients were followed up for a mean period of 23 months (range, 2-65 months). All 25 patients confirmed that their palmar sweating resolved postoperatively, with both palms equally dry. Of the 25 patients, 20 (80%) complained of compensatory hyperhidrosis, which also was bilaterally symmetric. The areas involved were trunk (80%), lower limbs (32%), and armpits (12%). Overall, 80% of the patients were very satisfied with the procedure. The remaining 20% experienced mild to moderate compensatory hyperhidrosis, which did not seem to affect their lifestyle. CONCLUSION: The findings show that T2 ablation in thoracoscopic sympathectomy for palmar hyperhidrosis is as effective as T2-T3 ablation in terms of symptomatic relief, recurrence, compensatory hyperhidrosis, and patient satisfaction. PMID- 17404795 TI - New "sutureless" technique of ileostomy and colostomy. AB - BACKGROUND: Brooke ileostomy and colostomy are associated with infection in the stomal and/or main wounds, leading to complications. The source of infection is feces from the open lumen. The objective of the present study was to find a way to prevent infection and subsequent complications. It was started in 1986 by serendipity after discovery that if wound contamination from intestinal contents is stopped in the immediate postoperative period, by a complete small/large intestinal obstruction, infection and complications are prevented. This study was based on the principle of delayed-primary wound closure. METHODS AND PROCEDURES: Following its initial discovery for a Brooke ileostomy, the procedure was used for both ileostomy and colostomy. The stapled stoma was fixed to the opening in rectus sheath. It was covered with an appliance that has a transparent pouch, allowing daily inspection. During the period of postoperative ileus, there is angiogenesis on the serosal surface, making it refractory to infection. The stoma tends to bulge with the appearance of peristalsis. Stoma was then opened with electrocautery in a bedside procedure. The mucosal cuff protrudes, everts, advances with peristalsis, and "grafts" itself on angiogenesis on the surface of a single layer of serosa. The advancing margin of the mucosal cuff fuses with the circumference of the opening in dermis. The maturation of the stoma is natural and automatic. Absence of sutures reduced the tissue trauma and foreign body reaction, resulting in better wound healing. This new procedure was named "delayed-primary self-maturation" (DPSM). RESULTS: Thirty seven colostomies and nine ileostomies were performed using DPSM. Infection in the stomal and/or main wound and subsequent complications were prevented. CONCLUSIONS: Delayed-primary self-maturation is technically easier and more scientific than a conventional ileostomy or colostomy and is recommended for all types of stomas. PMID- 17404796 TI - Comparison of intraabdominal pressures using the gastroscope and laparoscope for transgastric surgery. AB - BACKGROUND: The peroral transgastric endoscopic approach for intraabdominal procedures appears to be feasible, although multiple aspects of this approach remain unclear. This study aimed to measure intraperitoneal pressure in a porcine model during the peroral transgastric endoscopic approach, comparing an endoscopic on-demand insufflator/light source with a standard autoregulated laparoscopic insufflator. METHODS: All experiments were performed with 50-kg female pigs under general anesthesia. A standard upper endoscope was advanced perorally through a gastric wall incision into the peritoneal cavity. The peritoneal cavity was insufflated with operating room air from an endoscopic light source/insufflator. Intraperitoneal pressure was measured by three routes: (1) through the endoscope biopsy channel, (2) through a 5-mm transabdominal laparoscopic port, and (3) through a 16-gauge Veress needle inserted into the peritoneal cavity through the anterior abdominal wall. The source of insufflation alternated between on-demand manual insufflation through the endoscopic light source/insufflator using room air and a standard autoregulated laparoscopic insufflator using carbon dioxide (CO(2)). RESULTS: Six acute experiments were performed. Intraperitoneal pressure measurements showed good correlation regardless of measurement route and were independent of the type of insufflation gas, whether room air or CO(2). On-demand insufflation with the endoscopic light source/insufflator resulted in a wide variation in pressures (range, 4-32 mmHg; mean, 16.0 +/- 11.7). Intraabdominal pressures using a standard autoregulated laparoscopic insufflator demonstrated minimal fluctuation (range, 8-15 mmHg; mean, 11.0 +/- 2.2 mmHg) around a predetermined value. CONCLUSION: Use of an on demand unregulated endoscopic light source/insufflator for translumenal surgery can cause large variation in intraperitoneal pressures and intraabdominal hypertension, leading to the risk of hemodynamic and respiratory compromise. Safety may favor well-controlled intraabdominal pressures achieved with a standard autoregulated laparoscopic insufflator. PMID- 17404797 TI - How does it feel? Sentinel node biopsy is better. PMID- 17404798 TI - Factors affecting influential discussions among physicians: a social network analysis of a primary care practice. AB - BACKGROUND: Physicians often rely on colleagues for new information and advice about the care of their patients. OBJECTIVE: Evaluate the network of influential discussions among primary care physicians in a hospital-based academic practice. DESIGN: Survey of physicians about influential discussions with their colleagues regarding women's health issues. We used social network analysis to describe the network of discussions and examined factors predictive of a physician's location in the network. SUBJECTS: All 38 primary care physicians in a hospital-based academic practice. MEASUREMENTS: Location of physician within the influential discussion network and relationship with other physicians in the network. RESULTS: Of 33 responding physicians (response rate = 87%), the 5 reporting expertise in women's health were more likely than others to be cited as sources of influential information (odds ratio [OR] 6.81, 95% Bayesian confidence interval [CI] 2.25-23.81). Physicians caring for more women were also more often cited (OR 1.03, 95% CI 1.01-1.05 for a 1 percentage-point increase in the proportion of women patients). Influential discussions were more frequent among physicians practicing in the same clinic within the practice than among those in different clinics (OR 5.03, 95% CI 3.10-8.33) and with physicians having more weekly clinical sessions (OR 1.33, 95% CI 1.15 to 1.54 for each additional session). CONCLUSIONS: In the primary care practice studied, physicians obtained information from colleagues with greater expertise and experience as well as colleagues who were accessible based on location and schedule. It may be possible to organize practices to promote more rapid dissemination of high-quality evidence-based medicine. PMID- 17404799 TI - Use of preventive services of overweight and obese Europeans aged 50-79 years. AB - BACKGROUND: Although frequent contacts with health care systems may represent more opportunities to receive preventive services, excess body weight has been linked to decreased access to preventive services and quality of care. OBJECTIVE: The objective of the study is to examine whether obese and overweight, compared to normal weight persons, have different experiences of preventive care. DESIGN: The study design is cross-sectional. Baseline data (2004) of a population-based survey conducted in 10 European countries. PARTICIPANTS: The participants were noninstitutionalized adults, 13,859, (50-79 years) with body mass index (BMI) > or = 18.5 kg/m2, who answered the baseline and supplementary questionnaires (overall response rate of 51.3%) of the Survey of Health, Ageing and Retirement in Europe (SHARE). MEASUREMENTS: BMI was divided into normal weight (BMI, 18.5 24.9 kg/m2), overweight (BMI, 25.0-29.9 kg/m2), and obesity (BMI >30 kg/m2). Reported dependent variables were: influenza immunization, colorectal and breast cancer screening, discussion and recommendation about physical activity, and weight measurement. We performed multivariate logistic regressions, adjusting for age, sex, education, income, smoking, alcohol consumption, physical activity, and country. RESULTS: Overweight and obesity were associated with higher odds of receiving influenza immunization but not with receipt of breast or colorectal cancer screening. Overweight and obese individuals mentioned more frequently that their general practitioner discussed physical activity or checked their weight, which was not explained by chronic diseases or the number of ambulatory care visits. CONCLUSIONS: These first data from SHARE did not suggest that overweight or obesity were associated with decreased use of preventive services. PMID- 17404801 TI - Anthraquinone polyamines: novel channel blockers of N-methyl-D-aspartate receptors. AB - Polyamines, in particular spermine, as well as some natural and synthetic polyamine derivatives have been found to be blockers of N-methyl-D-aspartate receptors. We developed novel, polyamine-based channel blockers to analyze the structure of NMDA receptors. Anthraquinone polyamines block NMDA receptors with some selectivity compared to other glutamate receptors. Results using mutant NR1 and NR2 subunits identified amino acid residues that influence blockade by anthraquinone polyamines. The head group (anthraquinone) may be positioned at the selectivity filter/narrowest constriction of the channel and the polyamine tail penetrates this constriction into the inner vestibule below the level of the selectivity filter. The results are consistent with other work showing that NR1 (Asn616) and NR2B (Asn616), but not NR2B (Asn615), make the narrowest constriction of NMDA channel, and that the M3 segments from the two subunits, which form the outer vestibule, are likely staggered relative to each other in the vertical axis of the channel. PMID- 17404802 TI - Ubiquitin dependent and independent protein degradation in the regulation of cellular polyamines. AB - Protein degradation mediated by the ubiquitin/proteasome system is the major route for the degradation of cellular proteins. In this pathway the ubiquitination of the target proteins is manifested via the concerted action of several enzymes. The ubiquinated proteins are then recognized and degraded by the 26S proteasome. There are few reports of proteins degraded by the 26S protesome without ubiquitination, with ornithine decarboxylase being the most notable representative of this group. Interestingly, while the degradation of ODC is independent of ubiquitination, the degradation of other enzymes of the polyamine biosynthesis pathway is ubiquitin dependent. The present review describes the degradation of enzymes and regulators of the polyamine biosynthesis pathway. PMID- 17404803 TI - Polyamines and mRNA stability in regulation of intestinal mucosal growth. AB - The mammalian intestinal epithelium is a rapidly self-renewing tissue in the body, and its homeostasis is preserved through strict regulation of epithelial cell proliferation, growth arrest, and apoptosis. Polyamines are necessary for normal intestinal mucosal growth and decreasing cellular polyamines inhibits cell proliferation and disrupts epithelial integrity. An increasing body of evidence indicates that polyamines regulate intestinal epithelial cell renewal by virtue of their ability to modulate expression of various genes and that growth inhibition following polyamine depletion results primarily from the activation of growth-inhibiting genes rather than a simple decrease in expression of growth promoting genes. In this review article, we will focus on changes in expression of growth-inhibiting genes following polyamine depletion and further analyze in some detail the mechanisms through which mRNA stability is regulated by RNA binding proteins. PMID- 17404804 TI - Cinnamomin: separation, crystallization and preliminary X-ray diffraction study. AB - Cinnamomin from Cinnamonum camphora seeds, a type II ribosome-inactivating protein that interferes with protein biosynthesis in mammalian cells, can induce the apoptosis of carcinoma cells and be used as an insecticide. A rapid and improved method has been developed for the extraction and purification of cinnamomin from camphora seed. Purification of cinnamomin is achieved with two successive steps of hydrophobic interaction chromatography carried out on a fast protein liquid chromatography (FPLC) system. Crystals suitable for X-ray diffraction analysis were obtained by vapor diffusion method. A complete data set at 2.8 A resolution has been collected. Data indexation and refinement indicate that the crystal is orthorhombic with space group P2(1)2(1)2(1) and unit cell dimensions a = 52.39 A, b = 126.33 A, c = 161.45 A. There are two molecules per asymmetric unit. Initial phasing by molecular replacement method yielded a solution, which will contribute to the structure determination. A molecular model will further the understanding of the mechanism of cinnamomin function. The latter will be combined with bio-informatics to facilitate the medical and other applications of cinnamomin. PMID- 17404805 TI - Infective delirium. PMID- 17404806 TI - Significance of local mobility in aggregation of beta-galactosidase lyophilized with trehalose, sucrose or stachyose. AB - PURPOSE: The purpose of this study is to compare the effects of global mobility, as reflected by glass transition temperature (T(g)) and local mobility, as reflected by rotating-frame spin-lattice relaxation time (T(1rho)) on aggregation during storage of lyophilized beta-galactosidase (beta-GA). MATERIALS AND METHODS: The storage stability of beta-GA lyophilized with sucrose, trehalose or stachyose was investigated at 12% relative humidity and various temperatures (40 90 degrees C). beta-GA aggregation was monitored by size exclusion chromatography (SEC). Furthermore, the T(1rho) of the beta-GA carbonyl carbon was measured by (13)C solid-state NMR, and T(g) was measured by modulated temperature differential scanning calorimetry. Changes in protein structure during freeze drying were measured by solid-state FT-IR. RESULTS: The aggregation rate of beta GA in lyophilized formulations exhibited a change in slope at around T(g), indicating the effect of molecular mobility on the aggregation rate. Although the T(g) rank order of beta-GA formulations was sucrose < trehalose < stachyose, the rank order of beta-GA aggregation rate at temperatures below and above T(g) was also sucrose < trehalose < stachyose, thus suggesting that beta-GA aggregation rate is not related to (T-T(g)). The local mobility of beta-GA, as determined by the T(1rho) of the beta-GA carbonyl carbon, was more markedly decreased by the addition of sucrose than by the addition of stachyose. The effect of trehalose on T(1rho) was intermediate when compared to those for sucrose and stachyose. These findings suggest that beta-GA aggregation rate is primarily related to local mobility. Significant differences in the second derivative FT-IR spectra were not observed between the excipients, and the differences in beta-GA aggregation rate observed between the excipients could not be attributed to differences in protein secondary structure. CONCLUSIONS: The aggregation rate of beta-GA in lyophilized formulations unexpectedly correlated with the local mobility of beta-GA, as indicated by T(1rho), rather than with (T-T(g)). Sucrose exhibited the most intense stabilizing effect due to the most intense ability to inhibit local protein mobility during storage. PMID- 17404807 TI - Learning from viruses: the necrotic bodies of tumor cells with intracellular synthetic dsRNA induced strong anti-tumor immune responses. AB - PURPOSE: Coaxing dead tumor cells to induce specific immune responses is an attractive tumor therapy. However, there continues to be a need for adjuvants that can promote the cross-presentation of the dead tumor cells to induce specific anti-tumor response. Viral dsRNA has multiple mechanisms to promote the cross-presentation of viral antigens in virus-infected cells. We propose to learn from viruses by generating dead tumor cells having synthetic dsRNA delivered inside them to allow the dsRNA to promote the cross-presentation of dead tumor cells. MATERIALS AND METHODS: Using synthetic dsRNA, poly(I:C), and the TC-1 cervical cancer model, we evaluated the extent to which the poly(I:C) can promote the necrotic bodies of TC-1 cells to induce specific anti-tumor immune response. The poly(I:C) was either simply mixed with the dead TC-1 cells or pre-loaded inside them. RESULTS: Immunization of tumor-bearing mice with the necrotic bodies of tumor cells admixed with poly(I:C) significantly inhibited the tumor growth. More importantly, immunization with the necrotic bodies having poly(I:C) pre loaded inside led to a significantly stronger anti-tumor response than when the necrotic bodies were simply admixed with the poly(I:C), apparently through a CD8(+) CTL response-mediated mechanism. CONCLUSIONS: These findings are expected to be clinically relevant for devising improved whole cell-based tumor vaccines. PMID- 17404809 TI - In vitro chondrogenesis of mesenchymal stem cells in recombinant silk-elastinlike hydrogels. AB - PURPOSE: In this study the chondrocytic differentiation and cartilage matrix accumulation of human mesenchymal stem cells (hMSCs) were investigated after encapsulation in a genetically engineered silk-elastinlike protein polymer SELP 47 K as an injectable matrix for delivery of cell-based therapeutics. MATERIALS AND METHODS: hMSCs were encapsulated in SELP-47 K and cultured for 4 weeks in chondrogenic medium with or without transforming growth factor-beta3 (TGF). Chondrogenic differentiation was evaluated by histological, RNA and biochemical analyses for the expression of cartilage extracellular matrix components. RESULTS: Histological and immunohistochemical staining revealed that the cells acquired a rounded morphology and were embedded in significant amounts of chondrogenic extracellular matrix. Reverse transcriptase (RT)-PCR showed an up regulation in aggrecan, type II and type X collagen and SOX9 in presence of TGF beta3. By day 28, constructs cultured in the presence of TGF-beta3 exhibited significant increase in sulfated glycosaminoglycan and total collagen content up to 65 and 300%, respectively. CONCLUSIONS: This study demonstrates that SELP-47 K hydrogel can be used as a scaffold for encapsulation and chondrogenesis of hMSCs. The ability to use recombinant techniques to precisely control SELP structure enables the investigation of injectable protein polymer scaffolds for soft-tissue engineering with varied physicochemical properties. PMID- 17404810 TI - Focused-ion-beam milling: a novel approach to probing the interior of particles used for inhalation aerosols. AB - PURPOSE: The current study aimed to examine the pharmaceutical applications of the focused-ion-beam (FIB) in the inhalation aerosol field, particularly to particle porosity determination (i.e. percentage of particles having a porous interior). MATERIALS AND METHODS: The interior of various spray dried particles (bovine serum albumin (BSA) with different degrees of surface corrugation, mannitol, disodium cromoglycate and sodium chloride) was investigated via FIB milling at customized conditions, followed by viewing under a high resolution field-emission scanning electron microscope. Two sets of ten particles for each sample were examined. RESULTS: For the spray-dried BSA particles, a decrease in particle porosity (from 50 to 0%) was observed with increasing particle surface corrugation. Spray-dried mannitol, disodium cromoglycate and sodium chloride particles were determined to be 90-100%, 0-10% and 0% porous, respectively. The porosity in the BSA and mannitol particles thus should be considered for the aerodynamic behaviour of these particles. CONCLUSIONS: The FIB technology represents a novel approach useful for probing the interior of particles linking to the aerosol properties of the powder. Suitable milling protocols have been developed which can be adapted to study other similar particles. PMID- 17404808 TI - Bile acid transporters: structure, function, regulation and pathophysiological implications. AB - Specific transporters expressed in the liver and the intestine, play a critical role in driving the enterohepatic circulation of bile acids. By preserving a circulating pool of bile acids, an important factor influencing bile flow, these transporters are involved in maintaining bile acid and cholesterol homeostasis. Enterohepatic circulation of bile acids is fundamentally composed of two major processes: secretion from the liver and absorption from the intestine. In the hepatocytes, the vectorial transport of bile acids from blood to bile is ensured by Na+ taurocholate co-transporting peptide (NTCP) and organic anion transport polypeptides (OATPs). After binding to a cytosolic bile acid binding protein, bile acids are secreted into the canaliculus via ATP-dependent bile salt excretory pump (BSEP) and multi drug resistant proteins (MRPs). Bile acids are then delivered to the intestinal lumen through bile ducts where they emulsify dietary lipids and cholesterol to facilitate their absorption. Intestinal epithelial cells reabsorb the majority of the secreted bile acids through the apical sodium dependent bile acid transporter (ASBT) and sodium independent organic anion transporting peptide (OATPs). Cytosolic ileal bile acid binding protein (IBABP) mediates the transcellular movement of bile acids to the basolateral membrane across which they exit the cells via organic solute transporters (OST). An essential role of bile acid transporters is evident from the pathology associated with their genetic disruption or dysregulation of their function. Malfunctioning of hepatic and intestinal bile acid transporters is implicated in the pathophysiology of cholestatic liver disease and the depletion of circulating pool of bile acids, respectively. Extensive efforts have been recently made to enhance our understanding of the structure, function and regulation of the bile acid transporters and exploring new potential therapeutics to treat bile acid or cholesterol related diseases. This review will highlight current knowledge about structure, function and molecular characterization of bile acid transporters and discuss the implications of their defects in various hepatic and intestinal disorders. PMID- 17404811 TI - Multidrug resistance protein 1 (MRP1) in rabbit conjunctival epithelial cells: its effect on drug efflux and its regulation by adenoviral infection. AB - PURPOSE: To evaluate the expression, localization, function, and regulation of multidrug resistance protein (MRP1) in rabbit conjunctival epithelial cells (RCEC). MATERIALS AND METHODS: MRP1 gene expression in RCEC was determined by reverse transcription-polymerase chain reaction (RT-PCR), and MRP1 protein expression and its localization were determined by Western blot analysis and immunofluorescence using an anti-MRP1 monoclonal antibody, MRPr1. The effect of MRP1 on the transport and uptake of fluorescein was evaluated in RCEC grown on Transwell filters. Moreover, the effect of adenovirus type 5 (Ad5)-infected RCEC, and cytokines (Interleukin 1 (IL-1), IL-6, and tumor necrosis factor alpha (TNFalpha)) on MRP1 expression and leukotriene C4 (LTC4) uptake were investigated. RESULTS: A 652 bp RT-PCR product from rabbit conjunctiva showed a 87% homology to human MRP1. Immunostaining with MRPr1 revealed a predominant basolateral localization of MRP1 in RCEC. Uptake of fluorescein, a MRP1 substrate, was increased (203-290%) in the presence of uricosuric drug probenecid at 100 microM, anti-inflammatory drug indomethacin at 10 microM and diclofenac, flurbiprofen, and ofloxacin at 1 mM, and by ATP depletion, but not influenced by the depletion of GSH, and the presence of antiviral cidofovir and anti inflammatory drug cromolyn and prednisolone. Apical-to-basolateral facilitated transport of LTC4 was abolished in the presence of probenecid. Western blot analysis with MRPr1 revealed a distinct band at approximately 190 kDa for freshly isolated and cultured RCEC. Both Ad5 and cytokines (IL-1, IL-6, and TNF-alpha) up regulated MRP1 expression, thereby reducing LTC4 uptake. CONCLUSIONS: MRP1 appears to be primarily localized in the basolateral membrane of RCEC and function in the efflux of certain organic anions and inflammatory factors out of cells from the basolateral membrane. The upregulation in the expression of MRP1 by Ad5-infection and cytokines suggests a role of MRP1 in the transport of inflammatory factors during ocular inflammation. Supported by NIH grants EY12578, EY10421, and EY12356. PMID- 17404812 TI - FDG-PET for pharmacodynamic assessment of the fatty acid synthase inhibitor C75 in an experimental model of lung cancer. AB - PURPOSE: Fatty acid synthase (FAS) is an emerging target for anticancer therapy with a variety of new FAS inhibitors being explored in preclinical models. The aim of this study was to use positron emission tomography with [(18)F]fluorodeoxyglucose (FDG-PET) to monitor the effects of the FAS inhibitor C75 on tumor glucose metabolism in a rodent model of human A549 lung cancer. MATERIALS AND METHODS: After a baseline FDG-PET scan, C75 was administered and post-treatment scans were performed serially. FAS activity was measured in treated animals ex vivo by [(14)C]acetate incorporation in animals euthanized in parallel to those imaged. RESULTS: Longitudinally measured metabolic volumes of interest and tumor/background ratios demonstrated a transient, reversible decrease in glucose metabolism and tumor metabolic volume after treatment, with the peak effect seen at 4 h. FDG-PET measurements correlated with changes in tumor FAS activity measured ex vivo. CONCLUSIONS: Because C75 causes an effect that is shorter in duration than expected, modification of the current weekly dosing regimen should be considered. These results demonstrate the utility of small animal FDG-PET in assessing the pharmacodynamics of new anticancer agents in preclinical models. PMID- 17404813 TI - Influence of mouthpiece geometry on the aerosol delivery performance of a dry powder inhaler. AB - PURPOSE: To investigate the influence of mouthpiece geometry on the amount of throat deposition and device retention produced using a dry powder inhaler (Aerolizer), along with the subsequent effect on the overall inhaler performance. MATERIALS AND METHODS: Computational Fluid Dynamics analysis of the flowfield generated in the Aerolizer with various modified mouthpiece geometries (including cylindrical, conical and oval designs) was used in conjunction with experimental dispersions of mannitol powder using a multi-stage liquid impinger to determine how the overall inhaler performance varied as the mouthpiece geometry was modified. RESULTS: Geometry of the inhaler mouthpiece had no effect on device retention or the inhaler dispersion performance. In contrast, the mouthpiece geometry strongly affected the amount of throat deposition by controlling the axial component of the exit air flow velocity. The radial motion of the emitted aerosol jet was found to have little effect on throat deposition in representative mouth-throat models. Despite the reduced throat deposition, there was no difference in the overall inhaler performance. CONCLUSIONS: For cases where low throat deposition is a key design parameter, this study demonstrates that the amount of throat deposition can be reduced by making minor modifications to the inhaler mouthpiece design. PMID- 17404814 TI - Novel L-Dopa and dopamine prodrugs containing a 2-phenyl-imidazopyridine moiety. AB - PURPOSE: The aim of this study was to gain insight into the feasibility of enhancing the delivery of L-Dopa and dopamine to the brain by linking these neurotransmitters and L-Dopa ethyl ester to 2-phenyl-3-carboxymethyl imidazopyridine compounds giving rise to the so-called Dopimid compounds. MATERIALS AND METHODS: A number of Dopimid compounds were synthesized and both stability and binding studies to dopaminergic and benzodiazepine receptors were performed. To evaluate whether Dopimid compounds are P-gp substrates, [(3)H]ritonavir uptake experiments and bi-directional transport studies on confluent MDCKII-MDR1 monolayers were carried out. The brain penetration properties of Dopimid compounds were estimated by the Clark's computational model and evaluated by investigation of their transport across BBMECs monolayers. The dopamine levels following the intraperitoneal administration of the selected Dopimid compounds were measured in vivo by using brain microdialysis in rat. RESULTS: Tested compounds were adequately stable in solution buffered at pH 7.4 but undergo faster cleavage in dilute rat serum at 37 degrees C. Receptor binding studies showed that Dopimid compounds are essentially devoid of affinity for dopaminergic and benzodiazepine receptors. [(3)H]ritonavir uptake experiments indicated that selected Dopimid compounds, like L-Dopa and dopamine hydrochloride, are not substrates of P-gp and it was also confirmed by bi directional transport experiments across MDCKII-MDR1 monolayers. By Clark's model a significant brain penetration was deduced for L-Dopa ethyl ester and dopamine derivatives. Transport studies involving BBMECs monolayers indicated that some of these compounds should be able to cross the BBB. Interestingly, the rank order of apparent permeability (P (app)) values observed in these assays parallels that calculated by the computational approach. Brain microdialysis experiments in rat showed that intraperitoneal acute administration of some Dopimid compounds induced a dose-dependent increase in cortical dopamine output. CONCLUSION: Based on these results, it may be concluded that some Dopimid compounds can be proposed as novel L-Dopa and dopamine prodrugs. PMID- 17404815 TI - The influence of tissue handling on the flavonoid content of the aquatic plant Posidonia oceanica. AB - In recent times, more and more studies have focused on flavonoids as biomarkers of environmental quality in aquatic plants, in particular, Posidonia oceanica (Linnaeus) Delile. It is therefore of interest to determine how different prehandling methods can affect flavonoid concentrations. The methods tested were (1) immediate extraction of fresh samples, (2) extraction after 48 hr chilling, (3) freeze-drying, and (4) oven drying. Chilling and freeze-drying considerably altered the quantity of flavonoids measured, but not their profile. The effect of oven drying was not significant. Chilling led to a loss of 57% of total (pro)anthocyanidins, 39% of total flavonols, and 48% of all simple flavonols (myricetin, quercetin, isorhamnetin, and kaempferol). Freeze-drying caused a loss of 71% of total (pro)anthocyanidins, 87% of total flavonols, and 95% of all simple flavonols. PMID- 17404816 TI - Inhibition of proteinase inhibitor transcripts by Leptinotarsa decemlineata regurgitant in Solanum lycopersicum. AB - One mechanism by which plants defend themselves against insect herbivores is the production of plant proteinase inhibitors, which can inhibit digestion in the midgut, thus affecting growth and survival. In this work, the effect of Colorado potato beetle (CPB) [Leptinotarsa decemlineata (Say)] regurgitant on Solanum lycopersicum defenses was investigated. When regurgitant from fourth-instar CPB was applied to wounded S. lycopersicum leaves, the wound-induced transcripts for the proteinase inhibitors pin1 and pin2 were reduced. Boiling the regurgitant abolished its ability to reduce the pin transcripts. Ultrafiltration of the regurgitant demonstrated that it contained a component between 10 and 30 kDa molecular weight that inhibited wound-induced pin1 and pin2 expression, suggesting that it may be a protein. This may represent a mechanism that the CPB has evolved to elude the plant's induced response to infestation. PMID- 17404817 TI - Browse quality in quaking Aspen (Populus tremuloides): effects of genotype, nutrients, defoliation, and coppicing. AB - The consequences of interactions among genetic, ontogenetic, and environmental factors for the quality of winter-dormant tissues as food for browsing herbivores is poorly understood. We conducted two sequential common garden studies to assess the impacts of intraspecific genetic variation, nutrient availability, prior defoliation, and ontogenetic stage on the chemical quality of winter-dormant tissue in quaking aspen (Populus tremuloides Michx.). In the first study, saplings of 12 aspen genotypes were grown under low and high soil nutrient conditions, with or without two successive seasons of defoliation. Quantity and quality of current year's twig growth were assessed. Twig production varied among genotypes and declined under low nutrient availability, but showed little response to prior defoliation. Chemical quality of sapling twigs varied substantially among genotypes, and in response to nutrient availability and prior defoliation. Overall, browse quality improved (nitrogen levels increased while phenolic glycoside and condensed tannin levels decreased) after defoliation. Growth and chemical variables exhibited low to moderate clonal repeatability (broad sense heritability) values. Our second study employed the same 12 genotypes, grown under high-nutrient conditions and with or without two seasons of defoliation. The trees were coppiced to produce root sprouts, which were chemically assessed 1 yr later. Rejuvenation via coppicing led to increased levels of nitrogen, phenolic glycosides (salicortin), and tannins in root sprouts, and the magnitude of change varied among aspen genotypes. Signatures of defoliation nearly 2 yr earlier persisted in terms of elevated levels of phenolic glycosides in root sprouts of previously defoliated trees. Aspen forests likely present browsing herbivores with chemically heterogeneous environments because of the interactions of genetic, ontogenetic, and environmental factors that vary over space and time. PMID- 17404818 TI - Biosynthesis of tiglic, ethacrylic, and 2-methylbutyric acids in a carabid beetle, Pterostichus (Hypherpes) californicus. AB - Tiglic, 2-methylbutyric, and ethacrylic acids are found in the pygidial gland defensive fluid of many carabid beetles. By injecting a deuterium-labeled precursor into the carabid beetle Pterostichus (Hypherpes) californicus, and analyzing the defensive fluid by gas chromatography/mass spectrometry, we were able to demonstrate that tiglic and ethacrylic acids are biosynthesized from isoleucine via 2-methylbutyric acid. Moreover, we observed that the injection of L-isoleucine induces an increased production of tiglic acid in P. californicus. A strong primary kinetic isotope effect was found to operate in the dehydrogenation step of 2-methylbutyric acid to tiglic and ethacrylic acids. Consequently, ethacrylic acid was found to preferentially accumulate the deuterium labeling from [2,3,4,4-(2)H(4)]isoleucine during our biosynthetic experiments. PMID- 17404819 TI - Chemical changes associated with the invasion of a Melipona scutellaris colony by Melipona rufiventris workers. AB - Wax constituents produced by worker bees and the chemistry of the nest batumen (mixture of wax, mud, and floral materials) in a Melipona scutellaris colony changed when it was invaded by Melipona rufiventris workers. Gas chromatography/mass spectrometry analyses showed that after invasion, the M. scutellaris workers of the invaded colony produced waxes with higher relative abundance of triacontanyl acetate and decreased the amounts of n-alkanes and n-9 alkenes. On the other hand, waxes from M. rufiventris workers displayed few changes. The change in the composition of the M. scutellaris waxes chemically differentiates that species from the M. rufiventris invader workers. Comparative analyses of batumens samples from pure and invaded colonies revealed greater amounts of terpenes and phenolic derivatives in the batumen from the invaded colony. This is the first report on the chemical characterization of batumens from stingless bees. PMID- 17404820 TI - A 4-alkyl-substituted analogue of guaiacol shows greater repellency to savannah tsetse (Glossina spp.). AB - The responses of Glossina morsitans morsitans Westwood to guaiacol (2 methoxyphenol), a mild repellent constituent of bovid odors, and seven analogues comprising 2-methoxyfuran, 2,4-dimethylphenol, 2-methoxy-4-methylphenol (4 methylguaiacol), 4-ethyl-2-methoxyphenol (4-ethylguaiacol), 4-allyl-2 methoxyphenol (4-allylguaiacol; eugenol), 3,4-methylenedioxytoluene, and 3,4 dimethoxystyrene were compared in a two-choice wind tunnel. The 4-methyl substituted derivative (2-methoxy-4-methylphenol) was found to elicit stronger repellent responses from the flies compared with guaiacol. None of the other analogues showed significant repellent effects on flies. 4-Methylguaiacol, guaiacol, and eugenol (which was included because of previous reports of its repellency against a number of arthropods) were further evaluated in the field with wild populations of predominantly Glossina pallidipes Austen. The presence of guaiacol or eugenol near odor-baited traps caused some nonsignificant reduction in the number of tsetse catches at relatively high release rates (approximately 50 mg/hr). In contrast, the 4-methyl derivative at three different release rates (2.2, 4.5, and 9.0 mg/hr) reduced trap catches of baited traps in a dose-response manner. At 10 mg/hr release rate, it reduced the catches of baited and unbaited traps by approximately 80 and approximately 70%, respectively. In addition, the compound not only reduced the number of tsetse attracted to natural ox odor (approximately 80%), but also had an effect on their feeding responses, reducing the proportion that fed on an ox by more than 80%. Our study shows that the presence of a methyl substituent at the 4-position of guaiacol enhances the repellency of the molecule to savannah tsetse and suggests that 4-methylguaiacol may represent a promising additional tool in the arsenal of techniques in trypanosomiasis control. PMID- 17404821 TI - Microwave-triggered chemiluminescence with planar geometrical aluminum substrates: theory, simulation and experiment. AB - Previously we combined common practices in protein detection with chemiluminescence, microwave technology, and metal-enhanced chemiluminescence to demonstrate that we can use low power microwaves to substantially increase enzymatic chemiluminescent reaction rates on particulate silvered substrates. We now describe the applicability of continuous aluminum metal substrates to potentially further enhance or "trigger" enzymatic chemiluminescence reactions. Furthermore, our results suggest that the extent of chemiluminescence enhancement for surface and solution based enzyme reactions critically depends on the surface geometry of the aluminum film. In addition, we also use FDTD simulations to model the interactions of the incident microwave radiation with the aluminum geometries used. We demonstrate that the extent of microwave field enhancement for solution and surface based chemiluminescent reactions can be ascribed to "lightning rod" effects that give rise to different electric field distributions for microwaves incident on planar aluminum geometries. With these results, we believe that we can spatially and temporally control the extent of triggered chemiluminescence with low power microwave (Mw) pulses and maximize localized microwave triggered metal-enhanced chemiluminescence (MT-MEC) with optimized planar aluminum geometries. Thus we can potentially further improve the sensitivity of immunoassays with significantly enhanced signal-to-noise ratios. PMID- 17404822 TI - Bacterial DNA induces the complement system activation in serum and ascitic fluid from patients with advanced cirrhosis. AB - Translocation of intestinal bacteria to ascitic fluid is, probably, the first step in the development of spontaneous bacterial peritonitis in patients with cirrhosis. Proteins of the complement system are soluble mediators implicated in the host immune response to bacterial infections and its activation has been traditionally considered to be an endotoxin-induced phenomenon. The aim of this study was to compare the modulation of these proteins in response to the presence of bacterial DNA and/or endotoxin in patients with advanced cirrhosis and ascites in different clinical conditions. Groups I and II consisted of patients without/with bacterial DNA. Group III included patients with spontaneous bacterial peritonitis and Group IV with patients receiving norfloxacin as secondary long-term prophylaxis of spontaneous bacterial peritonitis. Serum and ascitic fluid levels of endotoxin and truncated residues of the complement system were measured by ELISA. The complement system is triggered in response to bacterial DNA, as evidenced by significantly increased levels of C3b, membrane attack complex, and C5a in patients from Groups II and III compared with patients without bacterial DNA (Group I) and those receiving norfloxacin (Group IV). Gram classification did not further differentiate the immune response between patients within groups II and III, even though endotoxin levels were, as expected, significantly higher in patients with bacterial DNA from gram-negative microorganisms. The complement protein activation observed in patients with bacterial DNA in blood and ascitic fluid is indistinguishable from that observed in patients with spontaneous bacterial peritonitis and may occur in an endotoxin independent manner. PMID- 17404824 TI - Cerebrovascular events after discontinuation of rofecoxib treatment. PMID- 17404823 TI - The cancer cell's "power plants" as promising therapeutic targets: an overview. AB - This introductory article to the review series entitled "The Cancer Cell's Power Plants as Promising Therapeutic Targets" is written while more than 20 million people suffer from cancer. It summarizes strategies to destroy or prevent cancers by targeting their energy production factories, i.e., "power plants." All nucleated animal/human cells have two types of power plants, i.e., systems that make the "high energy" compound ATP from ADP and P( i ). One type is "glycolysis," the other the "mitochondria." In contrast to most normal cells where the mitochondria are the major ATP producers (>90%) in fueling growth, human cancers detected via Positron Emission Tomography (PET) rely on both types of power plants. In such cancers, glycolysis may contribute nearly half the ATP even in the presence of oxygen ("Warburg effect"). Based solely on cell energetics, this presents a challenge to identify curative agents that destroy only cancer cells as they must destroy both of their power plants causing "necrotic cell death" and leave normal cells alone. One such agent, 3 bromopyruvate (3-BrPA), a lactic acid analog, has been shown to inhibit both glycolytic and mitochondrial ATP production in rapidly growing cancers (Ko et al., Cancer Letts., 173, 83-91, 2001), leave normal cells alone, and eradicate advanced cancers (19 of 19) in a rodent model (Ko et al., Biochem. Biophys. Res. Commun., 324, 269-275, 2004). A second approach is to induce only cancer cells to undergo "apoptotic cell death." Here, mitochondria release cell death inducing factors (e.g., cytochrome c). In a third approach, cancer cells are induced to die by both apoptotic and necrotic events. In summary, much effort is being focused on identifying agents that induce "necrotic," "apoptotic" or apoptotic plus necrotic cell death only in cancer cells. Regardless how death is inflicted, every cancer cell must die, be it fast or slow. PMID- 17404825 TI - Design and rationale of the ARBITER 6 trial (Arterial Biology for the Investigation of the Treatment Effects of Reducing Cholesterol)-6-HDL and LDL Treatment Strategies in Atherosclerosis (HALTS). AB - BACKGROUND: Recent evidence on the use of statin therapy indicates the potential for ultra-low levels of LDL-C to provide greater protection from recurrent coronary heart disease events. Guidelines for the treatment of lipid disorders were revised to indicate that an LDL-C treatment goal of 70 mg/dl was optional (NCEP ATPIII). In these same guidelines, low levels of HDL-C are also suggested but not specifically proscribed as a target of therapy. Recently ARBITER 2 (Arterial Biology for the Investigation of the Treatment Effects of Reducing Cholesterol 2) has provided the first evidence of the potential of raising HDL-C with extended release niacin when added to statin monotherapy in secondary prevention. However, whether this approach would be superior to a strategy in which lower concentrations of LDL-C are targeted is unknown. MATERIALS AND METHODS: ARBITER 6-HALTS ( HDL and LDL Treatment Strategies) will be a randomized, parallel group, open-label study comparing HDL-C and LDL-C focused strategies of lipid treatments for their effects on atherosclerosis. Up to 400 subjects will be assigned to either intensified LDL-C lowering therapy with ezetimibe or HDL-C raising therapy with extended-release niacin. The primary endpoint is the mean change in the intima-media thickness of the common carotid artery after 14 months. Secondary endpoints include the change in lipid values and lipid subfractions, drug discontinuation due to adverse effects, change in quality of life, and a composite endpoint consisting of all major adverse cardiovascular events. CONCLUSION: ARBITER 6-HALTS will guide clinicians on whether a lipid treatment strategy of raising HDL-C or further LDL-C reduction is superior in the secondary prevention of coronary heart disease. PMID- 17404826 TI - Anti-inflammatory actions of quinapril. AB - OBJECTIVE: The role of angiotensin II (Ang-II) in inflammation and the mechanisms through which it exerts this role are explored. Signaling through angiotensin stimulation of inflammatory cells often amplifies inflammation. Formation of Ang II from tissue angiotensin-converting enzyme (ACE) has been shown to be of greater importance in the development and progression of inflammatory diseases than plasma ACE. CONCLUSION: Quinapril, which is a potent and selective inhibitor of both plasma and tissue ACE, has demonstrated anti-inflammatory properties in many disease states such as atherosclerosis, nephritis, scleroderma, diabetes and arthritis, and, thus, offers new therapeutic possibilities for disease treatment. PMID- 17404827 TI - Retrograde adaptive resonance theory based on the role of nitric oxide in long term potentiation. AB - Adaptive resonance theory (ART) demonstrates how the brain learns to recognize and categorize vast amounts of information by using top-down expectations and attentional focusing. ART 3, one member of the ART family, embeds the computational properties of the chemical synapse in its search process, but it converges slowly and is lack of stability when being applied in pattern recognition and analysis. To overcome these problems, Nitric Oxide (NO), which serves as a newly discovered retrograde messenger in Long-Term Potentiation (LTP), is introduced in retrograde adaptive resonance theory (ReART) model presented in this paper. In the presented model a novel search hypothesis is proposed to incorporate angle and amplitude information of an external input vector to decide whether the input matches the long-term memory (LTM) weights of an active node or not, and the embedded NO retrograde mechanism makes the search procedure a closed loop, which improves the stability and convergence speed of the transmitter releasing mechanism in a synapse. To make the model more adaptive and practical, a forgetting mechanism is built to improve the weights updating process. Experimental results indicate that the proposed ReART model achieves low error rate, fast convergence and self-organizing weights regulation. PMID- 17404828 TI - Could oxidative stress from psychosocial stress affect neurodevelopment in autism? PMID- 17404829 TI - Dismantling institutional racism: theory and action. AB - Despite a strong commitment to promoting social change and liberation, there are few community psychology models for creating systems change to address oppression. Given how embedded racism is in institutions such as healthcare, a significant shift in the system's policies, practices, and procedures is required to address institutional racism and create organizational and institutional change. This paper describes a systemic intervention to address racial inequities in healthcare quality called dismantling racism. The dismantling racism approach assumes healthcare disparities are the result of the intersection of a complex system (healthcare) and a complex problem (racism). Thus, dismantling racism is a systemic and systematic intervention designed to illuminate where and how to intervene in a given healthcare system to address proximal and distal factors associated with healthcare disparities. This paper describes the theory behind dismantling racism, the elements of the intervention strategy, and the strengths and limitations of this systems change approach. PMID- 17404830 TI - Brief screening instrument of posttraumatic stress disorder for children and adolescents 7-15 years of age. AB - The objective of this paper is to develop a brief screening instrument of posttraumatic stress disorder (PTSD) for young victims of natural disasters. Data were derived from flood victims in 1998 and 1999 in Hunan, China. A representative population sample of 6,852 subjects 7-15 years of age was selected. Among them, 6,073 (88.6%) were interviewed. Multistage sampling was used to select the subjects and PTSD was ascertained with Diagnostic and Statistical Manual of Mental Disorders: 4th Edition (DSM-IV). We randomly assigned 80% (4,851) of the study subjects to construct the screening instrument (construct model) and the remaining 20% (1,222) subjects to examine the model (validation model). Logistic regression analysis and receiver operating characteristics curves were utilized to select a subset of symptoms and cutoff point from the pre-structured questionnaires. A seven-symptom instrument for PTSD screening was selected. Scores of 3 or more on this instrument were employed to define positive cases of PTSD with a sensitivity of 96.9%, specificity 99.0%, positive predictive value (PPV) 82.6%, and negative predictive value (NPV) 99.8%. The brief screening instrument developed in this study is highly valid, reliable, and predictable. PMID- 17404831 TI - Trace elements and pollutants concentrations in shorebirds from Yeongjong Island, Korea in the East Asian-Australian migration flyways. AB - This study presents concentration levels of trace metals and pollutants (zinc, manganese, copper, lead, and cadmium) in tissues (livers, kidneys, muscles, and bones) of shorebirds from Yeongjong Island, Korea, in the East Asian-Australian migration flyways. Essential trace elements, zinc concentrations in kidneys, and copper concentrations in muscles significantly differed among shorebirds, but manganese concentrations did not differ in each tissue. We suggest that essential elements are within normal range and are maintained there by normal homeostatic mechanism. Lead concentrations in livers, kidneys, muscles, and bones were significantly different among shorebird species. Lead concentrations in livers of Kentish Plovers, Mongolian Plovers, Dunlins, and Great Knots were less than the toxic level, and lead concentrations in livers of Terek Sandpipers were at the background level. Cadmium concentrations in livers, kidneys, muscles, and bones did not vary among shorebirds, and concentrations of cadmium in livers and kidneys were at background level in all shorebirds. In livers of Dunlins from Yeongjong Island, lead and cadmium concentrations were higher than other locations previously reported. PMID- 17404832 TI - Enhancement of sodium current in NG108-15 cells during neural differentiation is mainly due to an increase in NaV1.7 expression. AB - It is well known that morphological and functional changes during neural differentiation sometimes accompany the expression of various voltage-gated ion channels. In this work, we investigated whether the enhancement of sodium current in differentiated neuroblastoma x glioma NG108-15 cells treated with dibutyryl cAMP is related to the expression of voltage-gated sodium channels. The results were as follows. (1) Sodium current density on peak voltage in differentiated cells was significantly enhanced compared with that in undifferentiated cells, as detected by the whole-cell patch clamp method. The steady-state inactivation curve in differentiated cells was similar to that for undifferentiated cells, but a hyperpolarized shift in the activation curve for differentiated cells was observed. The sodium currents of differentiated and undifferentiated cells were completely inhibited by 10(-7) M tetrodotoxin (TTX). (2) The only Na(V) mRNA with an increased expression level during neuronal differentiation was that for NaV1.7, as observed by real-time PCR analysis. (3) The increase in the level of NaV1.7 alpha subunit expression during neuronal differentiation was also observed by immunocytochemistry; in particular, the localization of NaV1.7 alpha subunits on the soma, varicosities and growth cone was significant. These results suggest that the enhancement of TTX-sensitive sodium current density in differentiated NG108-15 cells is mainly due to the increase in the expression of the TTX sensitive voltage-gated Na+ channel, NaV1.7. PMID- 17404833 TI - Increased membrane/nuclear translocation and phosphorylation of p90 KD ribosomal S6 kinase in the brain of hypoxic preconditioned mice. AB - Our previous studies have demonstrated that hypoxic precondition (HPC) increased membrane translocation of protein kinase C isoforms and decreased phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) in the brain of mice. The goal of this study was to determine the involvement of p90 KD ribosomal S6 kinase (RSK) in cerebral HPC of mice. Using Western-blot analysis, we found that the levels of membrane/nuclear translocation, but not protein expression of RSK increased significantly in the frontal cortex and hippocampus of HPC mice. In addition, we found that the phosphorylation levels of RSK at the Ser227 site (a PDK1 phosphorylation site), but not at the Thr359/Ser363 sites (ERK1/2 phosphorylated sites) increased significantly in the brain of HPC mice. Similar results were confirmed by an immunostaining study of total RSK and phospho-Ser227 RSK. To further define the cellular populations to express phospho-Ser227 RSK, we found that the expression of phospho-Ser227 RSK co-localized with neurogranin, a neuron-specific marker, in cortex and hippocampus of HPC mice by using double labeled immunofluorescent staining method. These results suggest that increased RSK membrane/nuclear translocation and PDK1 mediated neuron-specific phosphorylation of RSK at Ser227 might be involved in the development of cerebral HPC of mice. PMID- 17404834 TI - Voltage-gated calcium channels provide an alternate route for iron uptake in neuronal cell cultures. AB - Recent studies suggest that iron enters cardiomyocytes via the L-type voltage gated calcium channel (VGCC). The neuronal VGCC may also provide iron entry. As with calcium, extraneous iron is associated with the pathology and progression of neurodegenerative diseases such as Parkinson's and Alzheimer's disease. VGCCs, ubiquitously expressed, may be an important route of excessive entry for both iron and calcium, contributing to cell toxicity or death. We evaluated the uptake of (45)Ca(2+) and (55)Fe(2+) into NGF-treated rat PC12, and murine N-2alpha cells. Iron not only competed with calcium for entry into these cells, but iron uptake (similar to calcium uptake) was inhibited by nimodipine, a specific L-type VGCC blocker, and enhanced by FPL 64176, an L-VGCC activator, in a dose-dependent manner. Taken together, these data suggest that voltage-gated calcium channels are an alternate route for iron entry into neuronal cells under conditions that promote cellular iron overload toxicity. PMID- 17404835 TI - Role of cell cycle proteins in CNS injury. AB - Following trauma or ischemia to the central nervous system (CNS), there is a marked increase in the expression of cell cycle-related proteins. This up regulation is associated with apoptosis of post-mitotic cells, including neurons and oligodendrocytes, both in vitro and in vivo. Cell cycle activation also induces proliferation of astrocytes and microglia, contributing to the glial scar and microglial activation with release of inflammatory factors. Treatment with cell cycle inhibitors in CNS injury models inhibits glial scar formation and neuronal cell death, resulting in substantially decreased lesion volumes and improved behavioral recovery. Here we critically review the role of cell cycle pathways in the pathophysiology of experimental stroke, traumatic brain injury and spinal cord injury, and discuss the potential of cell cycle inhibitors as neuroprotective agents. PMID- 17404836 TI - Species differences in the selective inhibition of monoamine oxidase (1-methyl-2 phenylethyl)hydrazine and its potentiation by cyanide. AB - The potentiating effects of cyanide on the inhibition of rat liver mitochondrial monoamine oxidase-A & B and of ox liver mitochondrial MAO-B by pheniprazine [(1 methyl-2-phenylethyl)hydrazine] has been studied. Pheniprazine was shown to behave as a mechanism-based MAO inhibitor. For rat liver MAO-B, the initial non covalent step was characterized by dissociation constant (K (i)) of 2450 nM and the first-order rate constant (k (+2)) for the covalent adduct formation was 0.16 min(-1). As a reversible inhibitor it was selective towards rat liver MAO-A (K (i) = 420 nM) but the rate of irreversible inhibition of that enzyme was considerably slower (k (+2) = 0.06 min(-1)). MAO-B from ox liver more closely resembled MAO-A from the rat in sensitivity to reversible inhibition by pheniprazine (K (i) = 450 nm) but it was closer to rat liver MAO-B in rate of irreversible inhibition (k (+2) = 0.29 min(-1)). The K (i) values were significantly decreased in the presence of KCN but there was little effect on the k (+2) values. However, sensitivities of the different enzymes to KCN varied widely and considerably higher concentrations of KCN were required for this effect to be apparent with the rat liver mitochondrial MAO-A than with MAO-B from rat and ox liver. The kinetic behaviour of cyanide activation was consistent with partial (non-essential) competitive activation in all cases. PMID- 17404837 TI - Maternal lead exposure produces long-term enhancement of dopaminergic reactivity in rat offspring. AB - To determine the effect of prenatal lead exposure on brain monoaminergic systems, pregnant rats were given tap water containing 250 ppm lead acetate, for the duration of pregnancy, while tap water without lead (Pb(2+)) was substituted at birth. Control rats were derived from dams that consumed tap water during pregnancy, and had no exposure to lead afterwards. At 12 weeks after birth, Pb(2+) content of brain cortex was increased 3- to 4-fold (P < 0.05). At this time the endogenous striatal levels of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid were 19% lower in Pb(2+) exposed rats (P < 0.05), while there was no change in the striatal level of dopamine (DA), noradrenaline, 3,4 dihydroxyphenylglycol, serotonin (5-HT) and 5-hydroxyindoleacetic acid (HPLC/ED). Also there was no change in these monoamines and metabolites in the prefrontal cortex of Pb(2+) exposed rats. However, turnover of 5-HT in prefrontal cortex, as indicated by 5-hydroxytryptophan accumulation 30 min after acute treatment with the decarboxylase inhibitor NSD-1015 (100 mg/kg IP), was lower in the Pb(2+) exposed rats. In the striatum AMPH-induced (1 mg/kg IP) turnover of DA, evidenced as L-DOPA accumulation after NSD-1015, was increased to a lesser extent in the Pb(2+) exposed rats (P < 0.05). The nitric oxide synthase inhibitor 7 nitroindazole (10 mg/kg IP) attenuated the latter effect, indicating that neuronal NO mediates this AMPH effect, at least in part. Moreover, DA D(2) receptor sensitivity developed in Pb(2+) exposed rats, as evidenced by enhanced quinpirole-induced yawning activity and enhanced quinpirole-induced locomotor activity (each, P < 0.05). These findings indicate that ontogenetic exposure to lead can have consequences on monoaminergic neuronal function at an adult stage of life, generally promoting accentuated behavioral effects of direct and indirect monoaminergic agonists, and related to increased dopamine turnover in basal ganglia. PMID- 17404838 TI - MHP-133, a drug with multiple CNS targets: potential for neuroprotection and enhanced cognition. AB - MHP-133 is one of a novel series of compounds designed to target multiple brain substrates expected to have synergistic actions in the treatment of cognitive and neurodegenerative disorders such as Alzheimer's disease. The strategy was to develop compounds with multiple targets relevant for enhancing cognition and memory, but avoiding the serious side effects attributed to high potency cholinergic agonists. MHP-133 was shown to interact with subtypes of cholinergic, serotonergic, and imidazoline receptors and to weakly inhibit acetylcholinesterase activity. In vitro, the drug enhanced nerve growth factor (TrkA) receptor expression; it prevented excitotoxicity in a hippocampal slice preparation; and increased the secretion of soluble (non-toxic) amyloid precursor protein. MHP-133 also enhanced cognitive performance by rats and by non-human primate in tasks designed to assess working memory. The results of this study are consistent with the potential use of MHP-133 in the treatment of neurodegenerative disorders such as Alzheimer's disease. PMID- 17404840 TI - Effect of chronic administration of ethanol on the regulation of tyrosine kinase phosphorylation of the GABAA receptor subunits in the rat brain. AB - One of the many pharmacological targets of ethanol is the GABA inhibitory system, and chronic ethanol (CE) is known to alter the polypeptide levels of the GABA(A )receptor subunits in rat brain regions. In the present study, we investigated the regulation of the tyrosine kinase phosphorylation of the GABA(A) receptor alpha(1)-, beta(2)- and gamma(2)-subunits in the rat cerebellum, cerebral cortex and hippocampus following chronic administration of ethanol to the rats. We observed either down-regulation or no change in the tyrosine kinase phosphorylation of the alpha(1) subunit, whereas there was an up-regulation or no change in the case of beta(2)- and gamma(2)-subunits of the GABA(A) receptors depending on the brain region following chronic administration of ethanol to the rats. These changes reverted back to the control level following 48 h of ethanol withdrawal. These results suggest that tyrosine kinase phosphorylation of GABA(A )receptors may play a significant role in ethanol dependence. PMID- 17404839 TI - Brain iron toxicity: differential responses of astrocytes, neurons, and endothelial cells. AB - Iron accumulation or iron overload in brain is commonly associated with neurodegenerative disorders such as Parkinson's and Alzheimer's diseases, and also plays a role in cellular damage following hemorrhagic stroke and traumatic brain injury. Despite the brain's highly regulated system for iron utilization and metabolism, these disorders often present following disruptions within iron metabolic pathways. Such dysregulation allows saturation of proteins involved in iron transport and storage, and may cause an increase in free ferrous iron within brain leading to oxidative damage. Not only do astrocytes, neurons, and brain endothelial cells serve unique purposes within the brain, but their individual cell types are equipped with distinct protective mechanisms against iron-induced injury. This review evaluates iron metabolism within the brain under homeostatic and pathological conditions and focuses on the mechanism(s) of brain cellular iron toxicity and differential responses of astrocytes, neurons, and brain vascular endothelial cells to excessive free iron. PMID- 17404841 TI - Angiogenic expression profile of normal and neurofibromin-deficient human Schwann cells. AB - Peripheral nerve sheath tumors from individuals with Neurofibromatosis Type 1 (NF1) are highly vascular and contain Schwann cells which are deficient in neurofibromin. This study examines the angiogenic expression profile of neurofibromin-deficient human Schwann cells relative to normal human Schwann cells, characterizing both pro-angiogenic and anti-angiogenic factors. Conditioned media from neurofibromin-deficient Schwann cell lines was pro angiogenic as evidenced by its ability to stimulate endothelial cell proliferation and migration. Using gene array and protein array analysis, we found increased expression of pro-angiogenic factors and decreased expression of anti-angiogenic factors in neurofibromin-deficient Schwann cells relative to normal human Schwann cells. Neurofibromin-deficient Schwann cells also showed increased expression of several growth factor receptors and decreased expression of an integrin. We conclude that neurofibromin-deficient Schwann cells have dysregulated expression of pro-angiogenic factors, anti-angiogenic factors, growth factor receptors, and an integrin. These dysregulated molecules may contribute to the growth and progression of NF1 peripheral nerve sheath tumors. PMID- 17404842 TI - The failure in NGF maturation and its increased degradation as the probable cause for the vulnerability of cholinergic neurons in Alzheimer's disease. AB - This short review discusses the arguments to consider the dismetabolism of the pathway responsible for both the maturation and degradation of NGF as the culprit of vulnerability of the forebrain cholinergic system to the Alzheimer's disease neuropathology. This summary includes information regarding a novel metabolic cascade converting Pro-NGF to mature NGF in the extracellular space and its ultimate degradation by a metalloprotease. It also describes how this pathway is altered in Alzheimer's disease with the consequential CNS accumulation of proNGF and impairment in the formation of NGF along with increased degradation of this key trophic factor. This metabolic scenario in Alzheimer's disease should result in the failure of NGF trophic support to forebrain cholinergic neurons and thus explaining the vulnerability of these neurons in this neurodegenerative condition. PMID- 17404843 TI - Alterations in mRNA expression of myelin proteins in the sciatic nerves and brains of streptozotocin-induced diabetic rats. AB - Diabetic neuropathy is the most common complication of diabetes. We examined the levels and the mRNA expression of myelin proteins in the sciatic nerves and the brains of streptozotocin-induced diabetic rats. The diabetic rats exhibited a decrease in body weight, elevation of the blood glucose level and a decrease in motor nerve conduction velocity at 2 weeks after streptozotocin injection. In the sciatic nerves of diabetic rats, the level of P0 protein and its mRNA expression were markedly reduced at 20 weeks after the injection. In the brains, the levels of proteolipid protein and myelin-associated glycoprotein and their mRNA expression were selectively decreased at 20 weeks after the injection. This affected expression of myelin proteins was found even when no histological abnormalities were detectable. Considering the functional significance of myelin proteins, this impairment of protein expression is possibly involved in the pathogenesis of diabetic neuropathy, including that in brain disorders. PMID- 17404844 TI - Screening preferences of patients at familial risk of colorectal cancer. AB - Our primary objective was to assess the screening preferences of patients at familial risk of colorectal cancer. Asymptomatic subjects aged 18-75 with a single first-degree relative diagnosed with colorectal cancer (n = 48) or polyps (n = 52) were asked to identify a preferred screening strategy, test features influencing their choice, and level of interest in decision-making after reviewing a decision aid describing the pros and cons of currently recommended screening tests. Although both groups preferred colonoscopy, 40% of subjects with a family history of colorectal cancer and 48% of those with a family history of polyps preferred alternative strategies. Accuracy was the most commonly identified test feature influencing test preference. Most subjects (66%) felt that selection of screening test should be a patient dominant or shared process. We conclude that patients at familial risk of colorectal cancer have distinct screening preferences that often vary from current recommendations. PMID- 17404845 TI - Flare-up of nonalcoholic steatohepatitis after hepatectomy resulted in hepatic failure in a patient with type 2 diabetes mellitus. PMID- 17404846 TI - Geranylgeranylacetone induces cyclooxygenase-2 expression in cultured rat gastric epithelial cells through NF-kappaB. AB - Geranylgeranylacetone (GGA) effectively protects the gastric mucosa against noxious agents. The precise mechanisms underlying the gastroprotective actions of GGA are not known. To elucidate the precise mechanism of GGA, the effect of GGA treatment on COX-2 expression in rat gastric epithelial (RGM1) cells was investigated. We used a prostaglandin E2 (PGE2) enzyme-linked immunoassay kit and Western blot analysis to measure PGE2 production and COX-2 induction by GGA treatment in serum-starved RGM1 cells. Gel-shift assay, Western blot analysis, and a reporter assay were performed to determine which COX-2 promoter was involved in GGA-induced COX-2 expression. GGA treatment dose dependently increased COX-2 expression and PGE2 production. The nuclear factor (NF)-kappaB sites of the COX-2 gene promoter were critical for GGA-mediated COX-2 expression. GGA induces COX-2 expression and increases PGE2 production in serum-starved RGM1 cells via activation of the NF-kappaB sites of COX-2 gene promoters. PMID- 17404847 TI - Combination therapy in liver transplant recipients with hepatitis B virus without hepatitis B immune globulin. AB - INTRODUCTION: Conventional therapy to prevent HBV recurrence in liver transplant (LTx) recipients consists of Hepatitis B Immune Globulin (HBIg). The aim of this review is to investigate the safety and efficacy of converting HBIg and LAM therapy to ADV and LAM therapy. METHODS: A retrospective review involving all liver transplant patients with HBV maintained on HBIg and LAM therapy. Results collected included: gender, age, HBV serological and DNA status (COBAS AmpliScreen PCR-based testing). Serologic testing was done every three months. Patients were followed for drug reactions, therapy compliance, and immune suppression compliance. A cost benefit analysis was done for drug comparisons using United States currency values. RESULTS: Patient demographics included: Male (n=6), Female (n=4), mean age 44 years (range 33 to 65). The mean length of follow up since therapy conversion (from HBIg and LMV to ADV and LMV) was 21 months (range 16 to 25 months). Serological status at time of conversion revealed that DNA status remained negative in all patients, HBsAg negative in 10/10, HB eAg (+) (5/10) and HBeAb (+)(5/10). None of the patients experienced an increase in transaminases while on dual ADV and LAM therapy. All patients were maintained on immune suppression monotherapy (tacrolimus) at 7-9 ng/mL. All patients reported compliance with the dual therapy and that they experienced no drug related side effects. Mean yearly costs for ADV and LAM was 7,235.00 United States dollars (range 6,550.00 to 8,225.00); while mean monthly costs for HBIg and LAM; 9225.00 (range 7205.00 to 12005.00). CONCLUSION: The above results demonstrate beneficial effects of ADV and LAM in place of the current standard of HBIg and LAM therapy. Safety and short term results show nucleoside therapy is adequate at preventing HBV viral recurrence. Lastly, the economic benefit for ADV and LAM vastly outweighed the HBIg and LAM group. PMID- 17404848 TI - Postoperative complications and functional results after subtotal gastrectomy with Billroth II reconstruction for primary gastric cancer. AB - Subtotal gastrectomy is considered the preferred treatment for gastric cancer with antral location. The aim of this study was to assess the incidence of early postoperative complications and late functional results in patients who underwent subtotal gastrectomy with Billroth II reconstruction for primary gastric adenocarcinoma. The results of 310 patients were analyzed with regard to postoperative complications and death rates. Functional results as they relate to the gastric resection were evaluated in 195 disease-free patients. Of the 310 patients, 77 developed postoperative general and surgical complications (24.8%) and 13 consequently died (in-hospital mortality: 4.2%). Although infrequent (6 cases, 1.9%), anastomotic leak was the most serious complication (4 cases died during the postoperative phase). Considering functional results, weight loss continued for the first trimester after surgery, after which it stabilized. Loss of appetite was rarely observed; early after the operation the majority of patients were consuming a normal diet and regularly consumed less than five meals per day (83.6%). Dumping syndrome was uncommon and usually resolved within one year (12.3% at three months, 9.5% after one year, 5.2% after two years). On the other hand, postprandial abdominal fullness was frequently observed (43.1% at three months, 36.1% after one year, 21.3% after three years, and 16.5% after five years). Billroth II reconstruction after subtotal gastrectomy is associated with a limited risk of anastomotic complications. Anastomotic leak, although infrequent, is a life-threatening complaint and requires prompt recognition and aggressive surgical treatment. The incidence of late complications was low and the majority of patients recovered from them within one year after surgery, although the occurrence of postprandial abdominal fullness was not completely irrelevant. PMID- 17404849 TI - PPARbeta/delta protects against experimental colitis through a ligand-independent mechanism. AB - Peroxisome proliferator-activated receptors (PPARs) beta/delta and gamma have overlapping roles in the negative regulation of inflammatory response genes. Ligand activation of PPARgamma protects against experimental colitis in mice. PPARbeta/delta can negatively regulate inflammation and is highly expressed in the epithelial cells of the colon, therefore PPARbeta/delta may also have a role in experimental colitis. In these studies, colitis was induced by dextran sodium sulfate (DSS) treatment in wild-type and PPARbeta/delta-null mice, with and without the PPARbeta/delta specific ligand GW0742. PPARbeta/delta-null mice exhibited increased sensitivity to DSS-induced colitis, as shown by marked differences in body weight loss, colon length, colonic morphology, myeloperoxidase activity and increased expression of mRNAs encoding the inflammatory markers interferon gamma, tumor necrosis factor-alpha, and interleukin-6 compared to similarly treated wild-type mice. Interestingly, these differences were not affected by ligand activation of PPARbeta/delta in either genotype. These studies demonstrate that PPARbeta/delta expression in the colonic epithelium inhibits inflammation and protects against DSS-induced colitis through a ligand-independent mechanism. PMID- 17404850 TI - Alleviation of experimental ulcerative colitis with the synthetic peptide, F2A4-K NS (Fibratide). AB - Recombinant fibroblast growth factors (FGFs) maintain the integrity of the gut epithelium and reduce mucosal injury in experimental inflammatory bowel disease (IBD). Chemically synthesized FGF mimetics could potentially extend the utility of FGFs by tailoring them for optimal bioactivity and oral administration, for example. Here, F2A4-K-NS (Fibratide), a synthetic FGF mimetic peptide, alleviated dextran sulfate sodium (DSS)-induced ulcerative colitis in mice when delivered systemically and, to a lesser extent, orally. Intraperitoneal injection of Fibratide (1 or 5 mg/kg/day) ameliorated DSS-induced ulcerative colitis, resulting in reduced weight loss, decreased colon wall thickening, and increased colon length. Fibratide also improved epithelial integrity by reducing histological-detectable crypt damage and inflammation. Orally administered Fibratide (1 mg/kg/day) was also effective in ameliorating symptoms with effects generally similar to those of intraperitoneal injection. In vitro studies were conducted to help clarify how Fibratide might act in vivo. Fibratide exhibited a modest enhancement of epithelial cell proliferation. On the other hand, Fibratide doubled the rate of epithelial cells migration and restitution in a cell culture model of wound repair. Collectively, the results indicate that Fibratide reduced the severity of experimental ulcerative colitis and may be potentially useful in the treatment of IBD. PMID- 17404851 TI - Clinical significance of incidental colorectal wall thickening on computed tomography scan in African-American and Hispanic patients. AB - We sought to assess the significance of an incidental finding of colorectal wall thickening (CRWT) on computed tomography (CT) scan in African-American and Hispanic patients. We retrospectively reviewed charts of African-American and Hispanic patients from January 1994 to December 2005. Those patients were included in whom the colonoscopy was performed due to incidental CRWT on CT scan. Patients with a history or a family history of colorectal malignancy, inflammatory bowel disease, or colorectal surgery, with an incomplete colonoscopic examination, or <18 years of age were excluded. Endoscopic and pathological findings were abstracted. Thirty-two patients met the criteria. Endoscopic examination was abnormal in 21 (65.6%). The positive predictive value of CRWT for abnormal endoscopic examination was 65.6%. Abnormal endoscopic examination revealed diverticulosis in 9 (43%), erythematous mucosa in 8 (38%), polyps in 6 (29%), mass in 2 (9%), thickened folds in 1 (5%), and diverticulitis in 1 (5%). Histopathological findings revealed colitis in 7 (33%), adenoma in 4 (19%), hyperplastic polyps in 4 (19%), adenocarcinoma in 2 (9%), lymphoid aggregates in 2 (9%), melanosis coli in 1 (5%), and normal in 1 (5%) in the abnormal examination group. Abnormal endoscopic examination was found in 65.6% of patients. The prevalence of colitis, adenomas, and malignancy was high, therefore abnormal CRWT warrants further endoscopic evaluation. PMID- 17404852 TI - Anti-inflammatory effects of pravastatin on Helicobacter pylori-induced gastritis in mice. AB - Pravastatin, an HMG-CoA reductase inhibitor, exerts anti-inflammatory effects via several mechanisms including induction of endothelial nitric oxide synthase (eNOS). We investigated the effect of pravastatin on Helicobacter pylori-induced gastritis in mice. Mice with or without H. pylori infection received intraperitoneal pravastatin daily for 1 week. Expression of eNOS mRNA and tumor necrosis factor-alpha mRNA and myeloperoxidase activity in gastric tissue was determined. Myeloperoxidase activity was reduced in a dose-dependent manner by pravastatin, with activity inhibited by 53.5 and 73.7% at doses of 0.3 and 1 mg/kg, respectively. At a dose of 1 mg/kg, pravastatin reduced the level of tumor necrosis factor-alpha mRNA by 52.7%, while it did not affect eNOS expression. Pravastatin had no effects on these inflammatory parameters in uninfected mice. Pravastatin did not affect the viability of H. pylori. In conclusion, pravastatin exerts an anti-inflammatory effect on H. pylori-induced gastritis in mice without affecting eNOS expression. PMID- 17404853 TI - A synergistic antitumor effect of interleukin-2 addition with CD80 immunogene therapy for peritoneal metastasis of gastric carcinoma. AB - The co-stimulatory molecule CD80 is a ligand of CD28, which plays a key role in the induction of cell-mediated immune responses. Many tumors, including gastric cancer, decrease the expression of CD80, which results in the failure of immune recognition. We evaluated the effect of interleukin-2 addition combined with CD80 infection on the peritoneal metastasis in gastric cancer. CD80 infection combined with interleukin-2 addition significantly increased the activated cytotoxicity of mononuclear cells compared to CD80 gene infection and compared to the lacZ control group. In vivo, the survival of animals with intraperitoneal tumor was longest in those given CD80 infection with interleukin-2 addition (median survival, 46 days), followed by those given interleukin-2 (39 days), those given CD80 infection (37 days), and those given lacZ (29 days). These results suggest that interleukin-2 addition might contribute to improving the observed outcome of CD80 immunogene therapy in peritoneal metastasis of gastric carcinoma. PMID- 17404854 TI - Ruptured pancreaticoduodenal artery pseudoaneurysm with chronic pancreatitis presenting as recurrent upper gastrointestinal bleeding. PMID- 17404855 TI - Outcomes of cystic lesions in the pancreas after extended follow-up. AB - Incidental pancreatic cysts are frequently detected, but no appropriate management guidelines have been issued for their management because their natural history is relatively unknown. The aim of this study was to investigate their long-term clinical outcomes. Pancreatic cysts detected from 1998 to 2004 were retrospectively reviewed in patients followed up for more than 3 months. A total of 182 patients with incidental pancreatic cysts (initial mean size, 1.8+/-1.1 cm) underwent follow-up for an average of 35.4 months. An age>/=60 years was found to be associated with an increase in cyst size, with an odds ratio of 2.56. Two malignant cysts were found among 20 patients who underwent surgical resection during follow-up. Old age was also associated with the presence of a premalignant or malignant cyst (P<0.01). Although the majority of incidental pancreatic cysts show an indolent behavior, long-term, regular follow-up studies should be considered for all pancreatic cysts, especially in the elderly. PMID- 17404856 TI - Orlistat reverse fatty infiltration and improves hepatic fibrosis in obese patients with nonalcoholic steatohepatitis (NASH). AB - Nonalcoholic steatohepatitis (NASH) may cause progressive hepatic fibrosis, cirrhosis, and hepatocellular carcinoma. Treatment, thus far, has been restricted to diet and weight loss, but without compelling results. In this study we aimed to evaluate the efficacy of orlistat therapy in obese patients with NASH. Fourteen obese patients with NASH underwent liver biopsy prior to and subsequent to 6 months treatment with orlistat (120 mg tid). Hepatic fat extension was graded as normal, mild, moderate, or severe. Hepatic fibrosis was scored on a scale from 0 to 4, with 0 denoting no fibrosis and 4, cirrhosis. Portal inflammation was scored as 0-3, with 0 = normal, 1 = mild, 2 = moderate, and 3 = severe inflammation. Fourteen patients had NASH associated with diabetes, hyperlipidemia, or obesity. Orlistat reduced fatty infiltration in 10 patients (70%; P<0.01), 3 of whom had normal liver fat content after treatment. Orlistat improved inflammatory activity by 2 grades in 28% and by 1 grade in 50% of patients and effected no change in 22% of patients. Five patients (35%) returned to normal inflammatory activity. Orlistat improved hepatic fibrosis by 2 grades in three patients (21%) and by 1 grade in seven patients (50%). There was no change in four patients (28%). Orlistat lowered aminotransferases levels, total cholesterol, triglycerides and low-density lipoprotein, respectively. Insulin resistance index and malonyl dialdehyde levels improved significantly after orlistat therapy, whereas HbAic remained unchanged. In conclusion, in obese patients with NASH, liver fibrosis and inflammation improved after therapy with orlistat. PMID- 17404857 TI - Dietary glutamine supplementation prevents mucosal injury and modulates intestinal epithelial restitution following ischemia-reperfusion injury in the rat. AB - The aim of the present study was to evaluate the preventive effect of a 2-day oral glutamine supplementation against intestinal ischemia-reperfusion (IR) injury in a rat. Male Sprague-Dawley rats were divided into four experimental groups: sham rats underwent laparotomy, sham-GLU rats underwent laparotomy and were treaded with enteral glutamine (GLU) given in drinking water (2%) 48 hr before and following operation, IR rats underwent occlusion of both the superior mesenteric artery and the portal vein for 30 min followed by 24 hr of reperfusion, and IR-GLU rats were treated with enteral glutamine 48 hr before and following IR. Intestinal mucosal damage (Park's injury score), mucosal structural changes, enterocyte proliferation, and enterocyte apoptosis were determined 24 hr following IR. Sham-GLU rats demonstrated a lower rate of cell apoptosis in jejunum and ileum compared to sham animals. IR-GLU animals demonstrated a greater jejunal and ileal bowel and mucosal weight, mucosal DNA, villous height and crypt depth, and enterocyte proliferation index in ileum and a lower injury score grade in jejunum compared to IR-nontreated rats. In conclusion, pretreatment with oral glutamine prevents mucosal injury and improves intestinal recovery following IR injury in the rat. PMID- 17404858 TI - Epidemiology, pathogenesis, and management of Clostridium difficile infection. AB - Clostridium difficile infection is an important health problem worldwide and leads to increased morbidity and mortality, particularly among the elderly population. Antibiotics, especially those with a broad spectrum, often trigger the infection; hence the use of unnecessary antibiotics should be avoided. Mild to moderate cases respond to metronidazole or vancomycin. Severe cases may require bowel resection. Chronic relapsing cases require a prolonged course of antibiotics, immunoglobulin, probiotics, and, occasionally, feces enema. This review provides a comprehensive update on pathogenesis and management of Clostridium difficile infection for health professionals all over the world. PMID- 17404859 TI - Oxidative stress and pathogenesis of inflammatory bowel disease: an epiphenomenon or the cause? AB - Crohn's disease (CD) and ulcerative colitis (UC), known as inflammatory bowel disease (IBD), are fairly common chronic inflammatory conditions of the gastrointestinal tract. Although the exact etiology of IBD remains uncertain, dysfunctional immunoregulation of the gut is believed to be the main culprit. Amongst the immunoregulatory factors, reactive oxygen species are produced in abnormally high levels in IBD. Their destructive effects may contribute to the initiation and/or propagation of the disease. We provided an extensive overview on the evidences from animal and human literature linking oxidative stress to IBD and its activity. Moreover, the effects of antioxidant therapy on IBD patients in randomized, controlled trials were reviewed and the need for further studies elaborated. We also summarized the evidence in support for causality of oxidative stress in IBD. PMID- 17404860 TI - Development of hepatocellular carcinoma in a patient with Crohn's disease treated with azathioprine. PMID- 17404861 TI - Metallothionein expression in Helicobacter-infected pregnant mice and their fetuses and pups. AB - Metallothionein (MT) expression was investigated in pregnant mice infected with H. pylori or H. felis and their fetuses and pups. Mice, healthy or infected with H. pylori or H. felis (n = 18/group), were sacrificed 2 weeks after impregnation or 4 weeks postpartum. Pups were sacrificed as fetuses, after birth, or at ages 11 or 28 days. Whole fetuses, stomachs, small intestines, and livers were assayed for MT. MT was increased (P<0.05) by two- and threefold in fetuses from H. pylori and H. felis-infected mothers, respectively, compared to control fetuses. Stomach MT of H. felis-infected pregnant mice, and newborns and 28-day pups from H. felis-infected mothers, was elevated (P<0.05) twofold compared to that of control mice and pups. Liver MT was decreased (P<0.05) in H. felis-infected mice 4 weeks postpartum (18%) and in their 11-day (69%) and 28-day (53%) pups, while small intestinal MT was decreased in H. felis-infected pregnant mice (17%), H. felis-infected mice 4 weeks postpartum (19%), and their 11-day pups (35%), compared to control mice. H. felis infection altered MT levels of pregnant mice, their fetuses and pups, and mice postpartum, which may be a response to the marked inflammation. PMID- 17404862 TI - Ganglioneuromatosis: an unusual cause of ileal stricture mimicking Crohn's disease. AB - Ileal idiopathic forms of ganglioneuromatosis in adults are extremely rare and represent a challenging pathologic condition for the clinician. We present two cases of ileal ganglioneuromatosis consisting of stricturing lesions that mimicked clinical and radiologic features commonly observed in Crohn's disease patients with ileal involvement. PMID- 17404863 TI - Cutaneous necrotizing vasculitis associated with hepatitis C virus infection. PMID- 17404864 TI - Clinical features, serum interleukin-6, and interferon-gamma levels of 34 turkish patients with hepatoportal sclerosis. AB - Hepatoportal sclerosis (HPS) is a clinical disorder of obscure pathogenesis with a variable clinical profile. The aim of the study was to summarize the clinical features of Turkish patients with HPS and to measure the serum levels of interleukin (IL)-6 and interferon (IFN)-gamma to determine the T helper cell profile in the pathogenesis. The study was conducted on 34 HPS patients (17 men, 17 women; mean age at diagnosis, 27+/-10 years) and 15 healthy controls. The clinical features of HPS patients including demographics, clinical history, laboratory, and ultrasonography findings were summarized. Serum IL-6 and IFN gamma levels were measured by using commercially available enzyme-linked immunosorbent assay kits. Gastrointestinal bleeding was the most common dominant presenting symptom. Majority of the patients had preserved liver function tests. Serum triglyceride levels were decreased in 30%. Abdominal ultrasonography revealed well-demarcated bands of increased echogenicity surrounding the portal vein wall and sudden narrowing of the intrahepatic second-degree portal vein branches in all cases. Spontaneous shunts and/or collaterals were seen in 13 cases (37%). Extrahepatic portal vein thrombosis were seen in 7 (20%) patients after at least 5 years of disease duration. Serum levels of both IL-6 (median, 3.2 pg/mL) and IFN-gamma (median, 7.8 pg/mL) were significantly higher in HPS patients compared with the control group (median, 1 pg/mL). HPS has variable clinical profile in different geographic areas of the world. Both Th1 and 2 cells may have a role in the regulation of immune response and pathogenesis of the disease. PMID- 17404865 TI - Clostridium butyricum TO-A culture supernatant downregulates TLR4 in human colonic epithelial cells. AB - The present study was performed to examine whether probiotics affect Toll-like receptor 4 (TLR4) expression in human colonic epithelial cells. Culture supernatants or heat-killed bacteria of Bacillus mesentericus TO-A, Clostridium butyricum TO-A, and Streptococcus faecalis T-110 were applied to human colonic epithelial cells. Treatment with C. butyricum TO-A culture supernatant significantly reduced TLR4 mRNA level (x0.16), even in the presence of interferon gamma (IFN-gamma; x0.21) as compared with untreated controls. High-performance liquid chromatography analysis showed that C. butyricum TO-A supernatant contains formate, acetate, and butyrate. Interestingly, TLR4 mRNA was significantly suppressed (x0.15-x0.22) only when cells were treated with solutions containing butyrate. Electrophoretic mobility shift assay suggested that the binding affinity of PU.1 to the promoter region of the TLR4 gene was markedly inhibited when the cells were treated with butyrate. This study suggested that butyrate produced by C. butyricum TO-A downregulates TLR4 mRNA level in human colonic epithelial cells. PMID- 17404866 TI - High affinity of ecabet sodium for inflamed colonic mucosa in ulcerative colitis. PMID- 17404867 TI - Acute tubular injury associated with mesalazine therapy in an adolescent girl with inflammatory bowel disease. AB - Mesalazine is a first-line drug in pediatric inflammatory bowel disease, and is effective as primary treatment and maintenance therapy. It's usually well tolerated, but various side effects have been described. A 15-year-old female with ulcerative colitis developed polyuria, polydipsia, vomiting, and fatigue. She was receiving mesalazine (500 mg, thrice daily, p.o.) and prednisolone for 4 months. She was detected as acute tubular injury as she had dehydration, acidosis, hypostenuria, hematuria, proteinuria, low levels of potassium, uric acid and bicarbonate. These findings were attributed to interstitial nephritis as a side effect of mesalazine, however as renal biopsy was disapproved by the parents, it was not confirmed. After discontinuation of mesalazine her renal tubular functions improved. Potassium and phosphorus supplements were stopped after 7 months, although she had to continue bicarbonate supplementation. We conclude that regular renal screening is important in patients receiving 5-ASA therapy to prevent rare but serious complications, such as interstitial nephritis sometimes leading to chronic renal failure. PMID- 17404868 TI - Gastrointestinal complications in renal transplant patients: a large, single center experience. PMID- 17404869 TI - Laparoscopically resected foregut duplication cyst (bronchogenic) of the stomach. PMID- 17404870 TI - Pneumocystis pneumonia during combined therapy of infliximab, corticosteroid, and azathioprine in a patient with Crohn's disease. PMID- 17404871 TI - Retransplantation for de novo hepatocellular carcinoma in a liver allograft with recurrent hepatitis B cirrhosis 14 years after primary liver transplantation. PMID- 17404872 TI - Establishment and characterization of a high metastatic potential in the peritoneum for human gastric cancer by orthotopic tumor cell implantation. AB - The aim of this study was to establish an orthotopic implantation model with high metastasis of gastric cancer to the peritoneum which is more faithful to clinical metastasis. A human gastric carcinoma cell line, GC9811, was injected as a single cell suspension into the stomach of nude mice. The cells from some peritoneum metastatic foci were expanded in vitro and subsequently implanted to the stomach wall of nude mice. By repeating the in vivo stepwise selection method for four rounds and cloning culture, we obtained a cell line designated GC9811-P, which developed peritoneal metastasis in 13 of 13 (100%) of mice, compared with only 20% of those implanted with parental GC9811. The metastatic foci in the peritoneum showed essentially the same histological appearance as those induced by parental cells. Tumor cell growth of GC9811-P in vitro was faster than that of GC9811. Motility assays demonstrated higher motility of GC9811-P than of GC9811. The adhesive ability of GC9811-P cells to laminin was lower than that of GC9811 cells, whereas the ability of GC9811-P cells to adhere to fibronectin was significantly higher than that of parental cells. Differences between GC9811-P and their parental GC9811 cells were found in expression levels of various molecules by flow cytometric and western blot. The findings indicated that up regulation in the expressions of CD155, VEGF, syndecan-1, and syndecan-2 or down regulation in the expressions of IL-6 and E-cadherin play an important role in the peritoneal metastasis of human gastric carcinoma cells. The high-metastatic cell line appears to be useful for investigating the mechanisms of peritoneal metastasis and preventing peritoneal metastasis of human gastric cancer. PMID- 17404873 TI - Rapid response of Henoch-Schonlein purpura to corticosteroids: correlation between skin and gastric mucosal lesions. PMID- 17404874 TI - A large flat adenoma located on the pylorus ring successfully treated by endoscopic submucosal dissection. PMID- 17404875 TI - Alcohol dehydrogenase (ADH) isoenzyme activity in the sera of patients with Helicobacter pylori infection. AB - Human gastric mucosa contains three classes of alcohol dehydrogenase (ADH) isoenzymes: I, III, and IV. Various factors have been found to influence gastric ADH activity. One of them is Helicobacter pylori infection, which is associated with gastric mucosal injury and leads to a decrease in gastric ADH activity. The aim of the study was to assess the effect of H. pylori infection on the serum activity of ADH isoenzymes. Serum samples were taken from 35 patients with H. pylori infection and from 35 healthy subjects. For measurement of class I isoenzyme activity we employed the fluorometric method, with class-specific fluorogenic substrate (4-methoxy-1-naphthaldehyde). The activities of class III and IV ADH isoenzymes were measured by the photometric method with formaldehyde and with m-nitrobenzaldehyde as substrate, respectively. Total activity of ADH was measured by a photometric method with p-nitrosodimethylaniline. The total activities of ADH and class IV isoenzyme were significantly higher in sera of patients with H. pylori infection compared to healthy subjects. The serum activity of other tested isoenzymes of ADH did not differ significantly between infected and noninfected groups. We conclude that H. pylori infection of gastric mucosa is reflected in the serum by a significant increase in class IV and total ADH activity. PMID- 17404876 TI - Demonstration of low-regulatory CD25High+CD4+ and high-pro-inflammatory CD28-CD4+ T-Cell subsets in patients with ulcerative colitis: modified by selective granulocyte and monocyte adsorption apheresis. AB - Low-CD25(High+)CD4(+), a subset of regulatory CD25(+)CD4(+) T cells and high inflammatory CD28(-)CD4(+) T cells can exacerbate ulcerative colitis (UC). This study sought to investigate the frequency of CD25(High+)CD4(+) and CD28(-)CD4(+) T cells in patients with UC and the changes in these cells during Adacolumn granulocyte and monocyte adsorption apheresis (GMA). Subjects were 12 patients with active UC, 11 with quiescent UC, and 14 healthy volunteers (HVs). The mean clinical activity index was 15.7 +/- 2.2 in active UC and 4.5 +/- 1.1 in quiescent UC. Peripheral blood samples were stained with CD4, CD25, and CD28 antibodies for flow cytometry. Patients with active UC received GMA and blood samples were examined before and after the first GMA session. Patients with active UC (P < 0.04) or quiescent UC (P < 0.02) had a higher percentage of CD28( )D4(+)T cells compared with HVs, while the percentage of CD28(+)CD4(+) T cells was lower in both UC groups compared with HVs (P = 0.03 and P < 0.02). Patients with active UC had a lower percentage of CD25(High+)CD4(+)T cells compared with quiescent UC patients (P < 0.001). A significant increase in CD25(High+)CD4(+) T cells was associated with GMA (P < 0.03). Low CD25(High+)CD4(+) and high CD28( )CD4(+) are prominent features in UC. The increase in CD25(High+)CD4(+) T cells induced by GMA should contribute to improved immune function. Additional studies are warranted, since a low frequency of CD25(High+)CD4(+) (-) and a high frequency of CD28(-)CD4(+) (-) expressing T cells might be a predictor of clinical response to GMA. PMID- 17404877 TI - Hodgkin's disease presenting with chylous ascites and cavernous transformation of the portal vein. PMID- 17404878 TI - Endoscopic treatment for repeated arterial bleeding with ulcerative colitis. PMID- 17404879 TI - Minocycline toxicity requiring liver transplant. PMID- 17404880 TI - Ileocecal ulcer in central China: case series. AB - Diseases of the ileocecal junction are mostly ulcerative. The aim of this study was to investigate the value of colonoscopy on the diagnosis of the causes of ileocecal ulcer. Clinical manifestations, diagnosis, and endoscopic and histopathologic features of 52 hospitalized cases of ileocecal ulcer were retrospectively analyzed. In this study, the causes of ileocecal ulcer were intestinal tuberculosis (32.7%), Crohn's disease (19.2%), carcinoma of the cecum (17.3%), ulcerative colitis (UC; 13.5%), solitary ulcer of the colon (9.6%), lymphoma (5.8%), and leiomyoma (1.9%). The concordance rate of diagnosis at endoscopy to final diagnosis was 78.8%; the rate of endoscopic misdiagnosis was 21.2%. The concordance rate of clinical diagnosis before colonoscopy to the final diagnosis was 17.3%. Colonoscopy plays an important role in the diagnosis of the diseases that lead to ileocecal ulcer. However, colonoscopy may cause some erroneous diagnosis. To attain the correct diagnosis, endoscopic features, pathologic results, and the other clinical data should be analyzed together. PMID- 17404881 TI - Molecular analysis of the p53 gene in patients with intestinal metaplasia of the cardia and Barrett's esophagus: characterization by sequencing. PMID- 17404882 TI - Salt and stress synergize H. pylori-induced gastric lesions, cell proliferation, and p21 expression in Mongolian gerbils. AB - Our aim was to determine if salt and stress enhance Helicobacter pylori (Hp) lesions in Meriones unguiculatus. Two hundred seventy-eight pathogen-free gerbils were allocated to seven groups: Hp-Sydney strain (45), 8% higher-salt diet (38), stress (60% space reduction/water immersion; 36), Hp + salt (33), Hp + stress (34), N-methyl-N-nitro-N-nitrosoguanidine (34), and sham (58). Gerbils were sacrificed at 1 week (67), 12 weeks (73), 52 weeks (65), and 68 weeks (73). Sydney, Padova, and Lauren classifications were blindly used. Proliferation, p53, p21, and apoptosis were assessed. Follicular active gastritis (grade 2/3) was observed in 10% of Hp gerbils, 38% of Hp + salt gerbils, and 29% of Hp + stress gerbils at 52 weeks and 67%, 83%, and 43% at 68 weeks (P < 0.05). Heterotopic proliferative glands were identified in synergy groups from 52 weeks, with increases in their number and size by 68 weeks. Higher proliferative rates were observed in Hp+salt gerbils (P < 0.0001), and p21 overexpression in Hp+salt and Hp+stress gerbils (both P's < 0.0001), by 68 weeks, without p53 increases. We conclude that salt and stress synergize Hp damage and increase pseudo-invasive gland foci. PMID- 17404883 TI - Cannabinoid CB(1) receptor and gastric acid secretion. PMID- 17404884 TI - Molecular and clinical characteristics in 46 families affected with Peutz-Jeghers syndrome. AB - Germline mutations of the tumor suppressor gene LKB1/STK11 are responsible for the Peutz-Jeghers syndrome (PJS), an autosomal-dominant disorder characterized by mucocutaneous pigmentation, hamartomatous polyps, and an increased risk of associated malignancies. In this study, we assessed the presence of pathogenic mutations in the LKB1/STK11 gene in 46 unrelated PJS families, and also carried genotype-phenotype correlation in regard of the development of cancer in 170 PJS patients belonging to these families. All LKB1/STK11 variants detected with single-strand conformational polymorphism were confirmed by direct sequencing, and those without LKB1/STK11 mutation were further submitted to Southern blot analysis for detection of deletions/rearrangements. Statistical analysis for genotype-phenotype correlation was performed. In 59% (27/46) of unrelated PJS cases, pathogenic mutations in the LKB1/STK11 gene, including 9 novel mutations, were identified. The new mutations were 2 splice site deletion-insertions, 2 missenses, 1 nonsense, and 4 abnormal splice sites. Genotype-phenotype analysis did not yield any significant differences between patients carrying mutations in LKB1/STK11 versus those without mutations, even with respect to primary biliary adenocarcinoma. This study presents the molecular characterization and cancer occurrence of a large cohort of PJS patients, increases the mutational spectrum of LKB1/STK11 allelic variants worldwide, and provides a new insight useful for clinical diagnosis and genetic counseling of PJS families. PMID- 17404885 TI - Recurrent giant hemangioma causing severe respiratory distress. AB - Hemangioma is a common benign tumor of the liver that is usually asymptomatic. If >4 cm (giant hemangioma), it could present symptoms related to bleeding, thrombosis, consumptive coagulopathy, or adjacent abdominal organ compression. If symptomatic surgical treatment should be considered, liver resection as well as enucleation are considered. Recurrences after surgical resection are rare. We herein present a case of woman admitted to the emergency room for acute severe respiratory distress. She had undergone 2 surgical resections of liver hemangiomas. The respiratory syndrome, as showed by chest x-ray, computed tomography scan, and nuclear magnetic resonance imaging, was related to a recurrent giant multiple hemangioma, creating a prominent compression of right lung with left mediastinal shift and left heart dislocation. It resulted in pulmonary parenchymal compression associated with reduced chest wall compliance and increased pulmonary pressure. The patient was not eligible for surgical treatment because of the disease extension and her clinical conditions. She died 6 months later from respiratory insufficiency and hypercapnic coma. PMID- 17404886 TI - Effect of Dai-kenchu-to on gastrointestinal motility based on differences in the site and timing of administration. AB - The purpose of this study was to investigate the effects of the herbal medicine dai-kenchu-to on gastrointestinal motility based on differences in the administration site and timing. We sutured strain-gauge transducers to the stomach (three), duodenum (one), jejunum (one), ileum (one), and colon (two) and inserted indwelling tubes into the stomach, jejunum, and proximal colon of beagles. Dai-kenchu-to was administered to each site during the fasting or fed state. During the fasting state, the prokinetic effects of dai-kenchu-to were evident at all administration sites. The effects were attenuated during the fed state. With intracolonic administration, a contraction similar to the giant migrating contraction-like contraction was induced during the fasting and the fed state, and defecation occurred. Despite the differences in administration site and timing, no contraction complex appeared orad to the administration sites. These results indicate that the prokinetic effects of dai-kenchu-to differ with the site or timing of administration. PMID- 17404887 TI - Periampullary choledochoduodenal fistula associated with ampulla of Vater carcinoma. PMID- 17404888 TI - CARD15 genotype-phenotype relationships in a small inflammatory bowel disease population with severe disease affection status. AB - Inflammatory bowel disease (IBD; MIM# 266600) is subdivided on the basis of clinical findings as either Crohn's disease (CD), ulcerative colitis (UC), or indeterminate colitis (IC). Three previously described mutations within the IBD susceptibility gene CARD15 (R702W, G908R, 1007fs) increase susceptibility to CD with a terminal ileal and/or ileocolonic location and fibrostenosing behavior. We undertook an association study using 477 unrelated IBD patients (248 CD, 172 UC, 57 IC) and 104 population controls to determine whether these previously described associations could be replicated in a small, accurately phenotyped cohort. Case-control and family-based approaches were employed to analyze CARD15 mutant allele and haplotype data. Analyses were initially performed in unstratified IBD cohorts. The R702W mutant allele was associated with CD on case control analysis (q=0.036, P=.004), and 1007fs with CD on pedigree disequilibrium testing (P=.020). All 3 CARD15 mutations increased susceptibility to a variety of CD subphenotypic manifestations, including early-onset CD in individuals with a family history of IBD, and CD complicated by extraintestinal disease. We also present evidence to suggest that R702W may predispose to a more generalized form of CD. Additionally, we confirm that CARD15 mutations are associated with terminal ileal/ileocolonic, and to a lesser extent, fibrostenosing CD. PMID- 17404889 TI - Extremely elevated CA19-9 in acute cholangitis. AB - A 68-year-old women was admitted with the chief complaints of epigastralgia with radiation to the back and jaundice. Fever and chills were also noted. The laboratory data showed alkaline phosphatase, 245 U/L (normal range: 28-94); Direct Bilirubin, 5.29 mg% (0-0.4); Total Bilirubin, 8.4 mg% (0.2-1.4). Surprisingly, the serum CA19-9 was 5673.8 U/ml (normal value: <37). Abdominal sonography showed dilated common hepatic duct. MRCP demonstrated diffuse biliary tree dilatation with a very low signal intensity in distal common bile duct. Surgical intervention reveled a black stone at distal common bile duct and was successfully retrieved. After operation, the CA19-9 fell to 63.1 u/ml two weeks later and returned to normal (10.4 U/ml) one month later. PMID- 17404890 TI - Role of vortices in growth of microbubbles at mitral mechanical heart valve closure. AB - This study is aimed at refining our understanding of the role of vortex formation at mitral mechanical heart valve (MHV) closure and its association with the high intensity transient signals (HITS) seen in echocardiographic studies with MHV recipients. Previously reported numerical results described a twofold process leading to formation of gas-filled microbubbles in-vitro: (1) nucleation and (2) growth of micron size bubbles. The growth itself consists of two processes: (a) diffusion and (b) sudden pressure drop due to valve closure. The role of diffusion has already been shown to govern the initial growth of nuclei. Pressure drop at mitral MHV closure may be attributed to other phenomena such as squeezed flow, water hammer and primarily, vortex cavitation. Mathematical analysis of vortex formation at mitral MHV closure revealed that a closing velocity of approximately 12 m/s can induce a strong regurgitant vortex which in return can instigate a local pressure drop of about 0.9 atm. A 2D experimental model of regurgitant flows was used to substantiate the impact of vortices. At simulated flow and pressure conditions, a regurgitant vortex was observed to drastically enlarge micron size hydrogen bubbles at its core. PMID- 17404892 TI - NSAIDs and breast cancer recurrence in a prospective cohort study. AB - OBJECTIVE: We examined the association between NSAID use and breast cancer recurrence in a prospective cohort of 2,292 early-stage breast cancer survivors diagnosed from 1997 to 2000 participating in the Life After Cancer Epidemiology (LACE) Study. METHODS: From 2000 to 2002, mailed questionnaires were used to obtain information on aspirin, ibuprofen, and other NSAID use and subsequent breast cancer events. A total of 270 recurrences (local, regional, and distant disease and new primary breast cancers) were reported and verified by medical record review. Cox proportional hazard models were used to estimate rate ratios (RR) and 95% confidence intervals (CI), adjusting for age at diagnosis, race, cancer stage, tamoxifen treatment, chemotherapy use, body mass index, and cyclooxygenase-2 (COX2) inhibitor use. RESULTS: Current, regular use (at least three days per week at time of questionnaire administration) of ibuprofen (RR, 0.56; 95% CI, 0.32-0.98), but not aspirin (RR, 1.09; 95% CI, 0.74-1.61), was associated with a statistically significant decreased risk of breast cancer recurrence. The combination of ibuprofen and other non-aspirin NSAIDs such as naproxen and sulindac reflected a similar reduction in risk (RR, 0.56; 95% CI, 0.33-0.95). No association was found for the non-NSAID analgesic acetaminophen. CONCLUSION: Our findings provide support for an inverse association between current, regular ibuprofen use and breast cancer recurrence. PMID- 17404893 TI - Selection of entomopathogenic fungi for use in combination with sub-lethal doses of imidacloprid: perspectives for the control of the leaf-cutting ant Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae). AB - In the first part of this study, four isolates of the fungus Beauveria bassiana (Bals.) Vuillemin (LPP1, LPP2, CG05 and CG24) and one isolate of Metarhizium anisopliae (Metsch.) Sorokin (CG46) were tested against adult foragers of Atta sexdens rubropilosa. Ants were allowed to walk on filter paper discs, inside Petri dishes, previously impregnated with 1 ml of a conidia suspension (2 x 10(7) conidia ml(-1)), maintained at 80% RH and 26 degrees C for 24 h and subsequently, transferred to sterile Petri dishes, maintained at 23 degrees C, 80% RH, 24 h dark. The mean values of LT(50) for LPP2, LPP1, CG46, CG24 and CG05 were 3.5, 3.7, 3.8, 4.2 and 4.4 days, respectively. Control insects for all tests in this study showed less than 10% mortality. Experiments were carried out to test the toxicity of imidacloprid (IMI) to A. sexdens rubropilosa. Mortality was evaluated 10 days following a 24 h exposure to the insecticide. Percent mortality caused by 500, 200, 100 and 10 ppm IMI was 77.8, 56.7, 45.5 and 5.5 respectively. Insects treated with 10 ppm IMI were observed to have reduced locomotor activity 24 h after exposure to the insecticide. The LC(50) of IMI was 154.3 ppm. Subsequent tests were carried out to evaluate the combination of a sub-lethal dose of IMI (10 ppm) and infection by CG24 (1 x 10(7) conidia ml(-1)). Mortality due to fungal infection alone was 43.3%. Mortality of insects treated with IMI followed by exposure to the fungus was 64.3%. These results indicate that IMI significantly increases the susceptibility of ants to infection by B. bassiana CG24. PMID- 17404894 TI - Guttation droplets of Penicillium nordicum and Penicillium verrucosum contain high concentrations of the mycotoxins ochratoxin A and B. AB - Eight of eleven ochratoxigenic isolates of Penicillium nordicum and Penicillium verrucosum produced guttation droplets when grown on Czapek yeast extract (CYA) agar for 10-14 days at 25 degrees C. Parallel cultivation of one strain each of P. nordicum and P. verrucosum on malt extract agar demonstrated that higher volumes of exudate are produced on this agar. However, HPLC analyses revealed higher concentrations of ochratoxin A (OTA) and B (OTB) in droplets originating from cultures on CYA. For quantitative determination of the mycotoxin contents, triplicates of three isolates each of P. nordicum and P. verrucosum were grown as single spot cultures on CYA for up to 14 days at 25 degrees C. Guttation droplets were carefully collected between day 11 and 14 with a microliter syringe from each culture. Extracts from exudates and corresponding mycelia as well as fungal free agar were analyzed by HPLC for the occurrence of ochratoxin A (OTA) and B (OTB). Mean concentrations ranging between 92.7-8667.0 ng OTA and 159.7-2943.3 ng OTB per ml were detected in the guttation fluids. Considerably lower toxin levels were found in corresponding samples of the underlying mycelia (9.0-819.3 ng OTA and 4.5-409.7 ng OTB/g) and fungal free agar (15.3-417.0 ng OTA and 12.7-151.3 ng OTB/g). This is the first report which shows that high amounts of mycotoxins could be excreted from toxigenic Penicillium isolates into guttation droplets. PMID- 17404895 TI - Antioxidative enzymes and isozymes analysis of taro genotypes and their implications in Phytophthora blight disease resistance. AB - Assessment of the differential expression of antioxidative enzymes and their isozymes, was done in 30 day-old ex vitro raised plants of three highly resistant (DP-25, Jhankri and Duradim) and one highly susceptible (N-118) genotypes of taro [Colocasia esculenta (L.) Schott]. Antioxidative enzymes were assayed in the ex vitro plants, 7 days after inoculation with the spores (15,000 spores ml(-1) water) of Phytophthora colocasiae Raciborski to induce taro leaf blight disease. Uninoculated ex vitro plants in each genotype were used as control. The activity of superoxide dismutase (SOD) and guaiacol peroxidase (GPX) increased under induced blight condition when compared with control. Increase in antioxidative enzymes was more (67-92%) in the resistant genotypes than that (21-29%) of the susceptible genotype. The zymograms of SOD and GPX in the resistant genotypes, with pathogenic infection, showed increased activity for anodal isoform of SOD and increased expression and/or induction of either POX 1 or POX 2 isoforms of GPX. In susceptible genotype, expression of the above isoforms was faint for SOD and nearly absent for GPX under both blight free and induced blight conditions. Induction and/or increased activity of particular isoform of SOD and GPX against infection of Phytophthora colocasiae in the resistant genotypes studied led to the apparent conclusion of linkage of isozyme expression with blight resistance in taro. This might be an important criterion in breeding of taro for Phytophthora leaf blight resistance. PMID- 17404896 TI - Reliability, validity, and minimally important differences of the SF-6D in systemic sclerosis. AB - OBJECTIVES: To evaluate the reliability and validity and estimate the minimally important difference (MID) for the SF-6D in patients with systemic sclerosis (SSc). Subjects We used data from two clinical studies to analyze the SF-6D in patients with SSc: Study 1 was a cross-sectional observational study (N = 107) designed to assess three direct preference measures--the rating scale, time trade off, and standard gamble (SG) in patients with diffuse SSc and limited SSc, and Study 2 was a 12-month randomized, placebo-controlled, clinical trial (N = 168) assessing oral bovine collagen versus placebo in diffuse SSc. METHODS: We assessed the test-retest reliability of the SF-6D in Study 2 over a mean (SD) 4.8 (3.0)-week interval and the agreement between the SF-6D and direct preference measures in Study 1 using intraclass correlations (ICC). The MID was estimated using three different anchors--the SF-36 change in health item (patients who answered "somewhat better" formed the MID group), the Health Assessment Questionnaire-Disability Index (HAQ-DI; change of > or =0.14 and > or =0.22) and the skin score (change of > or =5.3). Results The mean (SD) SF-6D scores were 0.61 (0.12) in Study 1 and 0.64 (0.13) in Study 2. Test-retest reliability for the SF-6D was high (ICC = 0.82 [95% CI: 0.76, 0.87]). Agreement between the SF-6D and three direct preferences measures was poor to moderate (0.16-0.52). The MID estimate for the SF-6D using the change in SF-36 item -0.012 and this level of change was similar to the no change group. The mean MID estimate for the SF-6D improvement using the HAQ-DI and skin score as anchors was 0.035 (effect size of 0.27). CONCLUSION: This is the first study to assess the SF-6D in SSc. The SF-6D is reliable and valid in patients with SSc. We provide MID estimates that can aid in calculating sample size for clinical trials involving patients with diffuse SSc. PMID- 17404897 TI - Sense of coherence predicts change in life satisfaction among home-living residents in the community with mental health problems: a 1-year follow-up study. AB - OBJECTIVES: There is a call for a further investigation of Sense of Coherence (SOC), the central concept in salutogenesis, and its relation to health and life satisfaction. No previous studies have investigated the utility of SOC versus mental symptoms for the prediction of life satisfaction among people with chronic mental health problems (MHP). METHODS: The present study has a prospective design including a baseline assessment and a 1-year follow up. We recruited 107 adults from the community health care system. SOC was measured by the Sense of Coherence questionnaire, mental symptoms by the Symptom Checklist-90 revised and life satisfaction by The Quality of Life Scale (all Norwegian versions). RESULTS: The results show that while SOC predicts change in life satisfaction (standardized beta coefficient for SOC was 0.39, P = 0.014), mental symptoms did not (standardized beta coefficient 0.00, P = 1.0). CONCLUSIONS: These findings emphasize the importance of assessing factors that may explain differences in life satisfaction over and above mental symptoms among people with MHP. The results indicate that improving SOC among people with MHP might provide important opportunities for improving their life satisfaction. PMID- 17404898 TI - Predicting quality of life impairment in chronic schizophrenia from cognitive variables. AB - BACKGROUND: The aim of this study was to see whether and how cognition deficit predicts quality of life impairments in schizophrenia patients. METHOD: The Computerized Cambridge Automated Neuropsychological Test Battery, the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Quality of Life Scale (QLS) were used to assess 62 patients with chronic schizophrenia. Step-wise multiple regression analysis was used in order to determine cognitive variables that would predict the scores of each Q-LES-Q and QLS domain scores. RESULTS: Regression analysis revealed a significant association of the cognitive deficits with both general and domain-specific quality of life impairment measured with Q LES-Q and QLS. Deficits in executive functions, visual sustained attention, memory and motor skills have been found to be valid predictors both before and after controlling for the severity of symptoms, emotional distress, side effects, age, education, and illness duration. CONCLUSIONS: This study suggests that deficits in executive functioning, attention, memory and motor skills substantially contributes to predicting impairments across a wide range of HRQL domains, and, consequently, to quality of life appraisal in schizophrenia. Cognitive predictors cannot be attributed to illness-related and background variables. It can be concluded that, when aiming at the improvement of quality of life in schizophrenia patients, cognitive functioning should be targeted. PMID- 17404899 TI - Field testing of a European quality of life instrument for children and adolescents with chronic conditions: the 37-item DISABKIDS Chronic Generic Module. AB - OBJECTIVES: The aim of this study was to shorten the Health-Related Quality of Life (HRQL) DISABKIDS Chronic Generic Measure (DCGM) for children and adolescents and to test its reliability, construct, and external validity. STUDY DESIGN: 1153 children and adolescents (8-16 years) with chronic health conditions (asthma, arthritis, epilepsy, cerebral palsy, diabetes, atopic dermatitis, cystic fibrosis) and their family were recruited from different paediatric clinical settings in seven European countries. A two-time assessment comprised reports on sociodemographics, health status and HRQL of children/adolescents. RESULTS: The 37-item DCGM describes six dimensions (Independence, Physical Limitation, Emotion, Social Inclusion, Social Exclusion and Treatment) confirmed by Confirmatory Factor Analysis, multi-item scaling and item-goodness of fit to Rasch model. Internal consistency (Cronbach's alpha: 0.70-0.87) and test-retest reliability (ICC: 0.71-0.83) were satisfactory. Correlations between DCGM-37 and other HRQL instruments were the highest between dimensions evaluating similar concepts. Regarding discriminant validity of the DCGM-37, girls and older adolescents reported lower emotional we ll-being. Children belonging to families with low level of affluence and those with severe health conditions were found to have worse HRQL in all domains. CONCLUSION: Reliability, construct validity as well as convergent and discriminant validity of the DCGM-37 were shown. PMID- 17404900 TI - Measuring social difficulties in routine patient-centred assessment: a Rasch analysis of the social difficulties inventory. AB - BACKGROUND: Social difficulties may add to the psychological burden experienced by cancer patients. Therefore identifying social difficulties in routine oncology practice may help prevent or alleviate distress. The Social Difficulties Inventory (SDI) is a short questionnaire developed for assessing social difficulties in cancer patients. Although well-validated, not enough is known about the clinical meaning and utility of the instrument or whether the items can be meaningfully summed to form a summary index of "Social Distress". PURPOSE: To determine, using Rasch analysis, whether the SDI could be used as a summary index of social distress specifically examining three fundamental criteria: item fit, unidimensionality and item invariance. METHODS: The Partial Credit Model was applied to a heterogeneous sample of cancer patients (n = 609) who had completed the SDI. RESULTS: Five items were identified as misfitting (infit mean square > or = 1.3 and standardised t-statistic > or = 2) and excluded from the subsequent analysis. The remaining items formed a unidimensional interval scale with no additional factors identified in a principal components analysis of the residuals. No differential item functioning was observed for age, gender, diagnosis, extent of disease or social deprivation. The 16-item SDI can be summed to produce an overall index of social distress, facilitating routine identification of social difficulties. Subsequent work is needed to evaluate whether the instrument is able to identify patients with high levels of social distress requiring intervention. PMID- 17404901 TI - IL-4 inducibility in NKT cells, naive CD4+ T cells and TCR-gamma delta T cells. AB - NKT cells, naive CD4(+) T cells, and TCR-gammadelta T cells belong to distinct T cell lineages but all express T cell receptors generated through random combinatorial joining of V-(D)-J genes. These distinct lineage T cells also possess the property of promptly activating the IL-4 gene upon T cell receptor stimulation. A comparative accounting of features as they pertain to IL-4 inducibility in these three distinct lineage T cells is provided here. PMID- 17404902 TI - Chemistry, occurrence and biosynthesis of C-glycosyl compounds in plants. PMID- 17404903 TI - Tertiary indole alkaloids from leaves of Tabernaemontana dichotoma. AB - Seven indole alkaloids were isolated and identified from the leaves of Tabernaemontana dichotoma Roxb. The major alkaloids were: perivine, 19-epi voacristine, 12-methoxy-voaphylline and vobasine. (-)apparicine, 19-epi iboxygaine and isomethuenine occurred in minor amounts. PMID- 17404904 TI - Constituents of cultivated mulberry tree. AB - From the ethyl acetate extract of the cultivated mulberry tree (Morus alba L.), two new flavonoid derivatives with a fused dihydrochalcone partial moiety were isolated, and named kuwanons K and L. The structures of kuwanons K and L were shown to be 1 and 2 respectively, on the basis of chemical and spectral data. Kuwanons K ( 1) and L ( 2) are regarded biogenetically as Diels-Alder adducts of a chalcone derivative and a dehydroprenylflavonoid derivative, and are the first example of a flavonoid derivative possessing a cyclohexene ring at a B ring of a flavonoid derivative. PMID- 17404905 TI - [Sesquiterpenlactone aus Arnica chamissonis]. AB - From the flowers of Arnica chamissonis Less, sub-sp. foliosa (Nutt.) Maguire and its variety incana (Gray) Hulten two new naturally occuring pseudoguaianolides were isolated and their structures established by spectroscopic methods. They are shown to be 4-O-acetyl-6-desoxychamissonolide ( 3) and the propionylic ester of 11,13-dihydrohelenalin which was named arnicolide G ( 6). PMID- 17404906 TI - [8-epi-Valerosidate, a new iridoid glucoside from Penstemon serrulatus.]. AB - From leaves of Penstemon serrulatus Menz. a new esteriridoid has been isolated. The structure was established by spectroscopic methods (IR, MS, (13)C-NMR) and mainly (1)H-NMR-spectroscopy as 8-epivalerosidate. PMID- 17404907 TI - [Production of essential oils by cell-suspensions of Matricaria chamomilla in a two phase system.]. AB - Cell suspensions of Matricaria chamomilla were cultured in a two phase system consisting of an aqueous nutrient medium and a nontoxic lipophilic phase (triglyceride). Especially during the first week of culture period a great number of lipophilic substances were accumulated in the triglyceride phase as was demonstrated by TLC and GC. One of these substances could be identified as alpha Bisabolol using GC-MS-coupling. Cell growth was unimpaired by the addition of the lipophilic phase to the nutrient medium. PMID- 17404908 TI - [Time course of growth, evolution of sugar-nitrogen metabolism and accumulation of alkaloids in a cell suspension of Catharanthus roseus.]. AB - Intra and extra-cellular accumulation of three alkaloid markers, the disappearance of major nutrients from the medium and the evolution of phosphates, nitrates, ammonium ions, glucose and starch in the cells were observed during a time course study using a suspension of Catharanthus roseus. The accumulation of alkaloids is a complex phenomenon, not entirely unconnected with growth, which involves diffusion into the medium and cellular catabolism. It is difficult to relate the phenomenon to the appearance of disappearance of any one metabolite in particular. PMID- 17404909 TI - [Quantitative and qualitative HPLC-analysis of indole alkaloids from Catharanthus roseus cell cultures.]. AB - HPLC separation conditions for the qualitative and quantitative analysis of indole alkaloids from Catharanthus roseus tissue cultures were discribed as well as sample preparation procedures with Extrelut and Bond Elut SCX cartridges, which allow a quicker and more effective sample clean up as the classical alkaline chloroform extraction method. The methods are suitable for very small amounts of cell material. PMID- 17404910 TI - Effects of auxins on growth and saponin production in callus cultures of Panax ginseng. AB - A callus was induced from the petiole of Panax ginseng C. A. Meyer by treatment with 2,4-D. The callus has been maintained on the synthetic media containing 2,4 D as growth factor. Habituated ginseng callus which required no auxin for its growth was induced from a 2,4-D callus by successive treatments with a low concentration of 2,4-D. The fact that the process of habituation is reversible suggests that its mechanism is based upon adaptation rather than rare mutation followed by selection. It was also shown that the capacity of saponin production is significantly lower in habituated callus than in normal callus. Habituated callus which was maintained on 2,4-D media gradually recovered its capacity for saponin production and the degree of this capacity seemed to parallel the degree of its requirement for auxin. PMID- 17404911 TI - Phytochemical assessment of Lathyrus species for the neurotoxic agent, beta-N oxalyl-L-alpha-beta-diaminopropionic acid. AB - Lathyrus species have potential as feed crops, but many contain a neurotoxin, beta-N-oxalyl-L-alpha-beta-diaminopropionic acid (ODAP). An extraction and Spectrophotometric assay procedure are described for analysis of ODAP in small samples (0.5 g) of seed. The assay utilizes o-phthalaldehyde as a reagent, following hydrolysis of the ODAP. It is sensitive and specific, and suitable for the assay of 3-4 seeds of most Lathyrus species. The method has been used to screen a collection of Lathyrus accessions for use in a breeding program to develop agronomic strains with low toxicity. PMID- 17404912 TI - Bisbenzylisoquinoline alkaloids from Andrachne cordifolia. PMID- 17404913 TI - Scopolamine production in suspension cultures and redifferentiated roots of Hyoscyamus niger. AB - Adventitious roots were induced from 2-year-old, hyoscyamine rich suspension cultures of Hyoscyamus niger L. The cultured roots grew well in Linsmaier-Skoog medium containing 10 (-8)M 6-benzyladenine with 3% sucrose and produced much more scopolamine (0.12-0.30% dW) than hyoscyamine (0.04-0.08% dW). The alkaloids were detected in the medium as well as in the cells. Ten to twenty percent of the hyoscyamine added to the medium was converted to scopolamine within 24 days by the root cultures, but only 1 to 3% was converted by the suspension cultures. Scarcely any of the scopolamine added to the medium was converted to hyoscyamine by either culture. PMID- 17404914 TI - Regulation of saponin production in callus cultures of Panax ginseng [1]. AB - It was shown that Panax ginseng callus produces the same kinds of saponins in considerable amounts as in intact plants, and the saponin content is significantly lower in habituated callus than in normal callus. Furthermore, it was found that semicarbazide and particularly thiosemicarbazide inhibit the production of phytosterols and promote the biosynthesis of saponins in the presence of mevalonic acid, also the biosyntheses of saponins and phytosterols are inhibited by the individual end products. PMID- 17404915 TI - Distribution of pyrethrins in oil glands and leaf tissue of Chrysanthemum cinerariaefolium. AB - Leaves of Chrysanthemum cinerariaefolium at various stages of development were found to contain differing amounts of pyrethrins. The variation corresponds to the number of oil glands per unit area of leaf surface. Leaf tissue was mechanically and enzymatically separated into lower and upper epidermis and mesophyll cells. Pyrethrins were found to be present in each layer, more pyrethrins were present in the epidermal layers of young leaves while the mesophyll cells were found to have higher pyrethrins in older, more mature cells. PMID- 17404916 TI - Statistical evaluation of a number of glandular lipid constituents in ferns of three Dryopteris species. AB - The occurrence of various hydrocarbon, fatty acid and terpenoid constituents in glandular cells in the rhizomes of three Dryopteris fern species has been evaluated by statistical treatment of quantitative data from gas chromatographic analysis. The results obtained suggest complicated interrelationships, presumably resulting from the excretion process, between representatives of the three types of compounds. The species investigated show differences as to these interrelationships. PMID- 17404917 TI - Ariensin, a new lignan from Bursera ariensis. AB - From acetone extracts of the exudate of the tree Bursera ariensis, a new bilignan, named ariensin, has been isolated and characterized as (2R,3R)-bis-2.3 (3.4-methylenedioxybenzyl-)-1.4-butanediol diacetate. The structure has been determined from UV, IR, (1)H-NMR and (13)C-NMR data of the natural product, the dibromo derivative and the hydrolysis product. The optical rotation of the latter allowed the determination of the absolute configuration. PMID- 17404918 TI - The furocoumarin composition of Pituranthos triradiatus. AB - Five furocoumarins were isolated by HPLC from the petroleum ether extract of shoots of Pituranthos triradiatus (Hochst. ex Boiss.) Aschers. et Schweif. (Umbelliferae). This species is commonly used by Beduins still practicing folk medicine in the Negev and Sinai Deserts. The furocoumarins were found to be: isoimperatorin, 5-(3-methyl-2-butenyloxy)-psoralen; cnidilin, 8-methoxy-5-(3 methyl-2-butenyloxy)-psoralen; imperatorin, 8-(3-methyl-2-butenyloxy)-psoralen; bergapten, 5-methoxypsoralen; and isopimpinellin, 5,8-dimethoxypsoralen. Some chemical and physical characteristics of these compounds, including RT values, were determined. PMID- 17404919 TI - Furanoeremophilanes in Senecio filaginoides and S. pinnatus. AB - Aerial parts of Senecio filaginoides yielded two new furanoeremophilane type sesquiterpenoids: 6alpha-acetyloxy-10betaH-furanoeremophil-1-one, 1 and 6alpha tiglinoyloxy-10betaH-furanoerernophil-1-one, 2. Their structures have been elucidated by spectroscopic data. From roots and aerial parts of S. pinnatus 1alpha-hydroxy-6betaangeloyloxy-10alphaH-furanoeremophil-9-one, 3 has been isolated. PMID- 17404920 TI - A New benzopyranbenzofuran dimer from Encelia ventorum. AB - The chloroform extract of the stems of Encelia ventorum yielded a new benzopyranbenzofuran dimer. The structure was assigned from (1)H-NMR and MS data as 4[2-(5-acetyl-6-hydroxybenzofuranyl)]-2-[6(7-methoxy-2, 2-dimethylbenzo-1 pyranyl)]-pentan-2-ol. PMID- 17404921 TI - Isolation and structure of cleistanthoside A. AB - A new diphyllin diglycoside, named Cleistanthoside A has been isolated from the heartwood of Cleistanthus patulus Muell., A RG., and its structure elucidated as diphyllin-4-O-[beta- D-glucopyranosyl (1-->2)]-beta-3,4-di-O-methyl- D xylopyranoside. Other lignans isolated were (+)-sesamin, taiwanin C, paulownin, cleistanthin A and diphyllin, also beta-sitosterol. PMID- 17404922 TI - Alkaloids of Rauwolfia caffra seeds. AB - Rauwolfia caffra seeds yielded 0.012 per cent total alkaloids comprising the sarpagan compound normacusine B, alpha-yohimbine, alloyohimbine, an incompletely characterised yohimbine and the principal alkaloid, yohimbine. Chromatographic evidence indicated traces of norajmalan and ajmalan compounds. PMID- 17404923 TI - Alcaloides de l'Uncaria guianensis. AB - The alkaloidal content of the leaves, stems and root barks of U. guianensis has been studied. All compounds isolated are oxindole alkaloids; alkaloidal composition is compared to that of other South-American Uncaria species. PMID- 17404924 TI - A rapid procedure for the isolation of Catharanthine, vindoline and vinblastine. PMID- 17404926 TI - [peroxidase activity and peroxidase isoenzyme patterns of tissue cultures of Digitalis lanata and Digitalis purpurea.]. PMID- 17404925 TI - [The effect of aging on the formation of secondary products by tissue cultures of Digitalis purpurea.]. PMID- 17404927 TI - Research on african medicinal plants. PMID- 17404928 TI - Glucoacetosyringone from Ranzania japonica. PMID- 17404929 TI - Otosenine from Senecio aureus. PMID- 17404930 TI - Essential oil of Rabdosia melissoides. PMID- 17404931 TI - Kurt mothes 3.11.1900 - 12.2.1983. PMID- 17404933 TI - [Isolation and detection of phorbolesters in crotonoil with HPLC.]. AB - A HPLC method using reversed-phase columns is described for the determination and isolation of phorbolesters in crotonoil. The identification of the compounds is based on an off-line combination of HPLC-MS, as well as on fatty acid analysis by GC. With this method several new phorbolesters of crotonoil could be detected. This method is also suitable for the determination of phorbol- or similar diterpenesters in other Euphorbiaceae. PMID- 17404934 TI - Biotransformation of Cannabidiol to Cannabielsoin by Suspension Cultures of Cannabis sativa and Saccharum officinarum. AB - Suspension cultures of CANNABIS SATIVA L. and SACCHARUM OFFICINARUM L. have been shown to biotransform cannabidiol to Cannabielsoin under normal growth conditions. Both Cannabielsoin C-1 stereoisomers were produced. Mass spectral and chromatographic evidence were used in this identification. This is the first reported biologically catalyzed synthesis of Cannabielsoin as well as the first biotransformation of any cannabinoid by plant tissue culture systems. Possible mechanisms and significance of this conversion are discussed. PMID- 17404935 TI - Indole Alkaloid Formation and Storage in Cell Suspension Cultures of Catharanthus roseus. AB - The localization and storage of alkaloids were investigated in a low producing cell suspension culture of CATHARANTHUS ROSEUS(L.) G. Don. (Apocynaceae). Fluorescence microscopy and electron microscopy indicate alkaloid accumulation to occur inside the vacuoles of particular cells. These alkaloid storage cells exhibit a vacuolar pH of 3, while "normal" cells of a suspension culture have a vacuolar pH of about 5. Alkaloids are taken up in their unprotonated forms, trapped by protonation inside the vacuole and are accumulated there. The differentiation of alkaloid storage cells depends both on the cell line and the growth conditions and seems to be a prerequisite for the accumulation of alkaloids in cell suspension cultures of CATHARANTHUS ROSEUS. PMID- 17404936 TI - Polyploid Complexes within the Genus Galium. AB - In this paper the structure elucidation of a new A-ring prenylated anthraquinone, called galiprenylin, and its isolation by means of preparative TLC from GALIUM ALBUM M ILL. (2n = 44) are described and discussed. PMID- 17404942 TI - Alkaloids of Litsea wightiana1. AB - Six aporphine alkaloids glaucine, boldine, norboldine, isoboldine, norcorydine and laurotetanine have been isolated from the ethanolic extract of the stems of LITSEA WIGHTIANA in which spasmolytic, hypothermic and blood pressure lowering activities have been confirmed. PMID- 17404943 TI - Liwaconitine, a New Diterpenoid Alkaloid from Aconitum forrestii. PMID- 17404944 TI - [Total Syntheses of Knightinol, Acetylknightinol and 2,3-Dihydrodarlingine; Tropane Alkaloids from Knightia strobilina and Bellendena montana.]. PMID- 17404945 TI - A New Flavanone from Adina cordifolia. PMID- 17404947 TI - Essential Oil from Pimpinella serbica Fruits. PMID- 17404948 TI - Studies on medicinal plants of sri lanka. PMID- 17404949 TI - Isolation of 5-hydroxy-7,4'-dimethoxyflavone from Gonystylus bancanus. PMID- 17404951 TI - Investigation of costus. PMID- 17404952 TI - Determination of aconitine alkaloids in aconitum roots. AB - A reliable method for extraction of the crude drug, Aconitum roots, and purification of the extract for its eventual separation and quantitation of the aconitine alkaloids by HPLC technique has been developed. The method comprises extraction of aconite powder first with ammoniacal ether and then with methanol (x 3 each), chromatography of the resultant extract over neutral alumina and elution with ethyl acetate-methanol (7:3) prior to HPLC. The method has been found to be reproducible and suitable for routine analysis of aconite and its pharmaceutical preparations. PMID- 17404953 TI - [Plant from New-Caledonia. Alkaloids of Melodinus reticulatus]. AB - (-)Tabersonine and (-)-11-methoxytabersonine were isolated from fruits of Melodinus reticulatus. Stems and leaves contain uvaol and eight alkaloids, five of which were previously described: (-)tabersonine, venalstonidine, kopsinine, venalstonine and its 3-oxoderivative. Three are new: 19-hydroxyvenalstonine, 19 hydroxyvenalstonidine and 3-oxovenalstonidine. PMID- 17404954 TI - Further alkaloids from bark of Fagara mayu. AB - In addition to the alkaloids canthin-6-one; dictamnine, chelerythrine, gamma fagarine, skimmianine, magnoflorine and 11-(2'-ketobutane)-dihydrochelerythrine, four further alkaloids were isolated from the bark of Fagara mayu (Bert. ex Hook et Arn.) Engler and edulinine, bocconoline, tembetarine were identified. A new base, N-methylpseudolaudanine, was also isolated. PMID- 17404955 TI - Seasonal variation of alkaloids of Adhatoda vasica and detection of glycosides and N-Oxides of vasicine and vasicinone. AB - The distribution pattern of the alkaloids of A. Vasica has been studied with change of season. The study also resulted in the detection of glycosides and N oxides of vasicine and vasicinone. PMID- 17404956 TI - Saponin production in cell suspension cultures of Panax ginseng. AB - Two high saponin-producing cell lines were selected from the original callus induced in 1967 and maintained in good state. For saponin production and growth in two cell lines, the comparison of static and suspension cultures, static cultures in the dark or in the light and rotary or reciprocal shaking cultures were made in detail. It was shown that the growth in rotary shaking cultures were 1.8 times higher than in reciprocal cultures, while the saponin production was almost the same, and the most effective hormonal condition was the combination of indole-3-butyric acid with kinetin. The saponin content per fresh weight was almost the same as that of the crude drug, whereas that per dry weight was 4 times higher. PMID- 17404957 TI - Selection of high pyrethrin producing tissue cultures. AB - Axenic callus and shoot cultures of pyrethrum have been shown to be capable of pyrethrin biosynthesis. The influence of explant source on the biosynthetic capacity in cultured tissue was examined. Explants of various plant organs were taken from high and low yielding plant selections. The conditions necessary for the establishment of these explants in culture are described. Analytical screening of the tissue lines enabled the selection of a few "high yielding" strains which were derived from high yielding plant selections. Differentiated cultures tended to produce more pyrethrins than did callus cultures. PMID- 17404958 TI - Production of Diosgenin in somatic callus tissues of Dioscorea floribunda. AB - Shoot-tip and tuber-derived callus tissues of Dioscorea floribunda Mart, and Gal. when grown in revised tobacco medium supplemented with 1 and 2 mg/12,4-D, contained 0.295% and 1.33% diosgenin, respectively. PMID- 17404959 TI - Association between berry characters and Solasodine content of solanum khasianum. AB - Correlation between number, grren weight, dry weight and solasodine percent of berry of the treated and control plant progenies in second generation raised from X-rayed (5 KR, 7.5 KR, 10 KR, 12.5 KR and 15 KR), EMS-treated (0.5%, 0.75% and 1% solution in PO (4)-buffer) and untreated dry seeds of Solanum khasianum Clarke revealed that only the numbers of large and small berries had significant correlation with solasodine percent. Correlation between induced variations (CV%) in these characters indicated that selection on berry characters through M (1) and M (2) generations was effective. Dispersion (due to mutagenesis) in the three berry characters have created the base for progressive selection for high solasodine yielders. PMID- 17404960 TI - Constituents of essential oil of Commiphora guidotti. AB - From the essential oil of Commiphora guidotti seven sesquiterpene hydrocarbons and a furanosesquiterpenoid, furanodiene, were isolated. PMID- 17404961 TI - Volatile constituents of Plectranthus rugosus leaf oil. AB - The essential oil of Plectranthus rugosus (Labiatae) was investigated by GLC, NMR and mass spectrometry. Main constituents are caryophyllene, germacrene D, alpha phellandrene, alpha-pinene, caryophyllene oxide, alpha-cadinol, delta-cadinene, limonene, beta-phellandrene, myrcene, and p-cymene. PMID- 17404962 TI - [Isolation of a Kawa-pyrone from Achyrocline satureioides]. AB - 6-(4'-Hydroxy-trans-styryl)-4-methoxy-2-Pyrone ( 1) has been isolated from overground parts including influrescences of Achyrocline satureioides D.C. with a yield of 0.02%. The structure was established by spectroscopic means and by comparing with a synthetic sample. PMID- 17404963 TI - On the molluscicidal activity of tannin containing plants. AB - The aqueous and methanolic extracts of a series of typical tannin containing plants exhibit strong molluscicidal properties against the freshwater snail biomphalaria glabrata, which is the intermediate host of schistosomiasis. The crude extracts of Krameria triandra (Krameriaceae) and Arctostaphylos uvaursi (Ericaceae) were active at concentrations of 50 ppm. Hydrolyzable and condensed tannins are responsable for the molluscicidal activity. This report indicates a new class of plant derived compounds which may have important practical application for the control of schistosomiasis in endemic areas. PMID- 17404964 TI - Structure of feruloylhistamine, a hypotensive principle of Ephedra roots. AB - From the crude drug "mao-kon", the underground part of Ephedra plant, we have isolated a new hypotensive imidazole alkaloid feruloylhistamine the structure of which has been determined as shown in formula I based on chemical and physical evidence. PMID- 17404968 TI - Clonal Multiplication of Rehmannia glutinosa. PMID- 17404969 TI - [Flavonols from Fruits of Vitex agnus castus]. PMID- 17404970 TI - Caffeic acid choline ester: a new plant constituent from Reseda luteola. PMID- 17404971 TI - High Performance Liquid Chromatographic Separation of Isoflavones and Structural Elucidation of Isoflavone 7-O-glucoside 6''-malonates from Cicer arietinum. AB - An efficient separation by HPLC of isoflavone aglycones and glucosides on reversed - phase columns is described. The main isoflavone constituents of CICER ARIETINUM L., various TRIFOLIUM species, BAPTISIA AUSTRALIS L. and ONONIS SPINOSA L. are formononetin 7-O-glucoside 6''-malonate and biochanin A 7-O-glucoside 6'' malonate. PMID- 17404972 TI - Chemistry, 13C-NMR Study and Pharmacology of Two Saponins from Colubrina asiatica. AB - From the leaves of COLUBRINA ASIATICA B RONGEN (Rhamnaceae) two saponins have been isolated and structurally elucidated, mainly by (13)C-NMR-spectroscopic methods, as jujubogenin-3-O-[2-O-acetyl-3-O-(3-O-beta-D-xylopyranosyl-4-O-acetyl beta-D-glucopyranosyl)-alpha-L-arabinoside] (colubrinoside) and jujubogenin-3-O- [2-O-acetyl -3-O- (2-O- beta -D- xylopyranosyl-beta-D-glucopyranosyl)-alpha-L arabinoside] (colubrin) respectively. Both saponins inhibit the spontaneous motility of mice, even at low doses (1 mg/ kg), they show an antagonistic effect on amphetamine and exert a synergistic activity on chlordiazepoxide. PMID- 17404973 TI - Biosynthesis of rutacridone: the N-methylation step. AB - The N-methylation of rutacridone was investigated. This dihydrofuroacridone derivative ist the main alkaloid in cell suspension cultures of RUTA GRAVEOLENS, strain R-20. The N-methyl group is provided by L-methionine. N-methylanthranilic acid serves as excellent precursor of rutacridone. Furthermore this particular precursor could be trapped after feeding anthranilic acid in short term experiments. PMID- 17404974 TI - Conversion of Elymoclavine to Paspalic Acid by a Particulate Fraction from an Ergotamine-Producing Strain of Claviceps sp. AB - The particulate fraction from the ergotamine-producing strain CLAVICEPS sp. PCCE1 catalyzed the conversion of [ (14)C]elymoclavine to paspalic acid. NADPH was required. Maximum conversion was 95%. Carbon monoxide (CO:0 (2), 4:1) and SK&F 525A (1.0 mM), cytochrome P-450 inhibitors, inhibited the conversion 94% and 50%, respectively. Minor amounts of paspalic acid (0.1 mg/1) were present in cultures. The particulate fraction from CLAVICEPS sp. SD 58, which accumulates elymoclavine in cultures, lacked activity for the conversion of elymoclavine to paspalic acid. PMID- 17404975 TI - Hybridisation for genetic improvement in the yield of diosgenin from fenugreek seed. AB - The inheritance of diosgenin in fenugreek seed was investigated using seed from parents, F (1) and F (2) generations grown in an individually randomised field experiment. Evidence based on GLC assay results for monohydroxysapogenin yield, indicates significant genetic segregation for this character in the two crosses examined. The potential exists for the genetic improvement of this and other characters in fenugreek using the hybridisation and assay methods described. PMID- 17404976 TI - [The Active Substances of Asarum europaeum.]. AB - The local anaesthetic activity of both phenylpropanoids of ASARUM EUROPAEUM trans isoasarone and trans-isoeugenol methyl ether were measured in comparison to the unsubstituted propenyl benzene and to the mono methoxy derivative trans-anethole resp. benzocaine, trans-Isosarone shows local anaesthetic activity with an intensity nearly equal to benzocaine. The quantitative structure-activity relationship in the series of phenylpropanoids has been analysed and established that the activity is increasing according to the number of methoxy groups. The results are compared with the lipophility and the surface activity of the substances. Surface activity changes parallel with the anaesthetic activity while the lipophility changes in contrary. PMID- 17404977 TI - Cultivation of Tripterygium wilfordii Tissue Cultures for the Production of the Cytotoxic Diterpene Tripdiolide. AB - The production of the cytotoxic agent tripdiolide employing plant tissue cultures of TRIPTERYGIUM WILFORDII is demonstrated. Detailed investigations of a developed cell line of T. WILFORDII in terms of tripdiolide production versus variations in growth conditions, for example media composition, are presented. Also the age and size of the inoculum and the incubation period with respect to tripdiolide formation are evaluated. PMID- 17404978 TI - New Constitutents of Roots of Polygonum cuspidatum. AB - A new naphtoquinone and 9 known phenolic components were isolated from a Japanese drug the roots of POLYGONUM CUSPIDATUM. The new component was proved to be 2 methoxy-6-acetyl-7-methyljuglone. PMID- 17404979 TI - A New Bitter Secoiridoid Glucoside from Gentiana scabra var. Buergeri. AB - A new strongly bitter acyl secoiridoid glucoside, scabraside, has been isolated along with two known secoiridoid glucosides, trifloroside and gentiopicroside (= gentiopicrin), from the rhizomes and roots of GENTIANA SCABRA var. BUERGERI. The structure of the scabraside was established by chemical and spectral evidence. PMID- 17404980 TI - Inhibition of butyrylcholinesterase activity by some isoquinoline alkaloids. AB - The inhibition of butyrylcholinesterase activity in human serum by protoberberine, benzophenanthridine and aporphine alkaloids as well as four model compounds has been studied. The mechanism of the inhibition is discussed on the basis of the different types of interaction of these compounds with butyrylcholinesterase and acetylcholinesterase. PMID- 17404982 TI - Infraspecific variation and alkaloids of papaver species. AB - Many species of PAPAVER exist as chemical races and, in order to illustrate this fact, examples have been chosen from the sections Miltantha and Oxytona. Future work on the alkaloids of PAPAVER species, including the possibility of novel compounds, aspects of biosynthesis and plant cell culture, are briefly discussed. PMID- 17404981 TI - Constituents of the Essential Oil of Artemisia coerulescens. AB - By combination of liquid-solid chromatography, capillary GLC and mass spectrometry, the essential oil obtained by steam distillation from the leaves and flowers of ARTEMISIA COERULESCENS L., growing in Tuscany (Poggio di Monselvoli, Siena), was analysed, alpha-Thujone, beta-thujone and camphor accounted for about 69% of the oil which contained more than a hundred compounds, most of which were present only in traces; 36 components were identified. PMID- 17404983 TI - [Quality and quantity of morphinan alkaloids detectable in interspecific hybrids of the genus papaver*]. AB - Viable hybrids were artificially produced between PAPAVER BRACTEATUM rich in thebaine on the one side and PAPAVER SOMNIFERUM, PAPAVER ORIENTALE and PAPAVER PSEUDO-ORIENTALE on the other side. Each of them is characterized by morphological and some physiological features as well as by its chromosome number. Biochemical analysis confirmed that P. SOMNIFERUM-P. BRACTEATUM hybrids contain very high morphine concentrations if the biosynthetic activity of the first parent is not to low. Also other conditions favouring the morphine yield of this hybrid could be elucidated. - Oripavine is the main alkaloid in P. ORIENTALE P. BRACTEATUM hybrids. However, its concentration remains low. This is probably due to the limited oripavine biosynthesis in P. ORIENTALE plants used for cross experiments. - P. BRACTEATUM-P. PSEUDO-ORIENTALE hybrids contain thebaine only in trace amounts. These small quantities are characteristic of the P. PSEUDO ORIENTALE parents. Interspecific hybridizations corresponding to a "genetic feeding experiment" generally result in increased concentrations of desired morphinan alkaloids. However, their effectiveness remarkably depends on the following fact: Low alkaloid levels may frequently be the expression of a reparable shortage, but they can also be caused by a strict regulation snowing dominant inheritance. PMID- 17404984 TI - Oripavine - a new opium alkaloid. AB - Oripavine was isolated from the dried capsules of a variety of PAPAVER SOMNIFERUM cultivated on Tasmania. PMID- 17404985 TI - Studies on the Synthesis of Morphine VI*. AB - Experiments to find a biomimetic route for synthesizing morphine are summarized. Systematic investigations on the phenolic oxidative coupling have been revised and evaluated. PMID- 17404986 TI - Partial Purification and Properties of (S)-Norlaudanosoline Synthase from Eschscholtzia tenuifolia Cell Cultures. AB - A new enzyme, (S)-norlaudanosoline synthase, which catalyses the synthesis of (S) norlaudanosoline from dopamine and 3,4-dihydroxyphenylacetaldehyde was isolated from the soluble protein extract of ESCHSCHOLTZIA TENUIFOLIA cell suspension cultures and purified approximately 40-fold. The apparent molecular weight of the enzyme is 15 500 Dalton. The pH optimum is 7.8, temperature optimum 40 degrees C, apparent K (M) values for dopamine and dihydroxyphenyl-acetaldehyde are 1.5 mM and 0.7 mM respectively. The synthase shows high substrate specificity in that only the phenylacetaldehydes are transformed but not the phenylpyruvates. No apparent cofactor requirement could be demonstrated. By means of isoelectric focusing and disc-gel electrophoresis evidence was obtained for the existence of four norlaudanosoline synthase isoenzymes, none of which catalyses the reaction of dopamine with 3,4-dihydroxyphenylpyruvate. These enzymes are responsible for the synthesis of (S)-norlaudanosoline, the key intermediate in the formation of isoquinoline alkaloids occurring in the plant kingdom. PMID- 17404987 TI - Characterization of polyneuridine aldehyde esterase, a key enzyme in the biosynthesis of sarpagine/ajmaline type alkaloids. AB - Polyneuridine aldehyde esterase (PNA-esterase) was isolated and partially purified from cell suspension cultures of RAUWOLFIA SERPENTINA B ENTH. The exceptionally high substrate specific enzyme catalyses the conversion of the monoterpenoid C (10-) into the C (9-)unit at the stage of polyneuridine aldehyde during the biosynthesis of sarpagine/ajmaline type alkaloids. The enzymatically formed compound is the new, labile indole alkaloid 16-epivellosimine. It is this alkaloid which functions as a branch point of the biogenetic routes leading to sarpagan and ajmalan type alkaloids. PNA-esterase occupies a key position in the biosynthesis of both alkaloid groups. PMID- 17404988 TI - Alkaloid distribution in some african rauvolfia species. AB - The distribution of indole alkaloids in the leaves, stems and roots of 10 African mainland RAUVOLFIA species is summarised and the interrelationships of the alkaloid types discussed. PMID- 17404989 TI - [Alkaloids from Annonaceae XLVII*: Alkaloides of Guatteria ouregou**]. AB - The Guianese Annonaceous plant GUATTERIA OUREGOU D UN. has yielded fourteen isoquinoline alkaloids, five of which are novel. They belong to several structural types; attention is specially drawn to three original skeletons, namely 7-hydoxy-7-methyl aporphinoids, 7- GEM-dimethyl aporphinoids and alpha- GEM-dimethyl cularinoids. The results concerning solvent shift in (1)H NMR spectra of some of these alkaloids are emphasized. Finally the phytochemical significance of the isolated alkaloids is discussed considering their chemotaxonomic and biogenetic interest. PMID- 17404990 TI - [Contributions to the Structure of Hypercratine, another Alkaloid of Ruspolia hypercrateriformis.]. AB - Besides the known pyrrolidine alkaloids ruspolinone ( 1), norruspolinone ( 2), and norruspoline ( 3) the "bispyrrolidine" alkaloid hypercratine was isolated from the roots of RUSPOLIA HYPERCRATERIFORMIS M. R. (Acanthaceae). After chemical and spectroscopic analysis of the new compound, structure 4 is suggested. PMID- 17404991 TI - Accumulation of quinolizidine alkaloids in plants and cell suspension cultures: genera lupinus, cytisus, baptisia, genista, laburnum, and sophora. AB - The patterns of quinolizidine alkaloids in cell cultures of 10 species of Fabaceae were analyzed by high-resolution GLC and GLC-MS and compared with the alkaloids present in the leaves of the respective plants. Lupanine was produced in all 10 cell suspension cultures as the main alkaloid. It was accompanied by sparteine, tetrahydrorhombifoline, 17-oxosparteine, 13-hydroxylupanine, 4 hydroxylupanine, 17-oxolupanine, and 13-hydroxylupanine esters as minor alkaloids in some species. The alkaloid patterns of the plants differed markedly in that alpha-pyridone alkaloids were the major alkaloids in the genera Cytisus, Genista, Laburnum and Sophora but were not accumulated in the cell cultures. These data further support the assumption that the pathway leading to lupanine is the basic pathway of quinolizidine alkaloids biosynthesis and that the other alkaloids are derived from lupanine. PMID- 17404992 TI - N-Methylation of Anthranilic Acid to N-Methylanthranilic Acid by Cell-free Extracts of Ruta graveolens Tissue Cultures. AB - Cell-free extracts from acridone synthesizing cell suspension cultures of RUTA GRAVEOLENS L. catalyze the N-methylation of anthranilic acid using S-adenosyl-L methionine as methyl donor. The stability of enzyme preparations was remarkably high during storage at -20 degrees C. Optimum activity was exhibited at pH 8.2, Mg (2+) was not required for maximum activity and EDTA did not affect the reaction rate. The rate of N-methylanthranilic acid formation was shown to be linear for about 45 min and was proportional to the protein concentration up to at least 0.350 mg of the enzyme preparation. In a number of suspension cultures of plant species not belonging to the Rutaceae this particular N methyltransferase was not found. Apparantly N-methylation of anthranilic acid is the first pathway-specific reaction in acridone alkaloid biosynthesis. PMID- 17404993 TI - Chemistry of Acronycine: III. AB - A trimer of noracronycine has been isolated from the mixture obtained after treating noracronycine with methanolic hydrochloric acid, and its structure deduced by partial synthesis of the dihydro derivative, and examination of the (1)H NMR spectral parameters in comparison with those obtained for the two dimers reported previously. PMID- 17404994 TI - Aziridines in Alkaloid Synthesis. IV. A Formal Synthesis of alpha-Dihydrocaranone and gamma-Lycorane. AB - A facile synthesis of the key intermediates for alpha-dihydrocaranone ( 17) and gamma-lycorane ( 18) has been achieved by employing the ring opening reaction of a N-benzylaziridine derivative ( 10). PMID- 17404995 TI - Alkaloids of Fumaria densiflora*. AB - FUMARIA DENSIFLORA DC., collected near Varna (Bulgaria) was investigated for its alkaloidal content. Protopine, cryptopine, coptisine, palmatine, adlumidiceine, (+/-)-sinactine, and a new spirobenzylisoquinoline alkaloid densiflorine were isolated. Benzophenanthridine alkaloids were not detected. PMID- 17404996 TI - Quaternary alkaloids of tinospora species. AB - The occurrence of quaternary alkaloids in TINOSPORA (and PARABAENA) species (Menispermaceae) has been studied. The main components were generally the protoberberine bases berberine and palmatine, with jatrorrhizine an occasional minor constituent, and the aporphine base magnoflorine. Choline was also often present. Only magnoflorine was detected in the PARABAENA material examined. PMID- 17404997 TI - [Monomeric Indole Alkaloids from Kopsia officinalis.]. AB - Ten monomeric indole alkaloids have been identified from the roots of KOPSIA OFFICINALIS. Four of them are known: (-)-kopsinine 1, (+)-5,22-dioxokopsane 2, ( )-tetrahydroalstonine 4, and (-)-quebrachamine 3; and six are new: (-) isoeburnamine 5 (enantiomer of the known alkaloid (+)-isoeburnamine), (-)-N methoxycarbonyl-12-methoxykopsinaline 7, (-)-N-methoxycarbonyl-11,12 methylenedioxy kopsinaline 8, (-)-N-methoxycarbonyl-11,12-dimethoxykopsinaline 9, (-)-11,12-methylenedioxykopsinaline 10, and (-)-12-methoxykopsinaline 11. PMID- 17405001 TI - Biotransformation by plant cells immobilized in cross-linked polyacrylamide hydrazide. AB - Plant cells were entrapped by mixing suspended MENTHA cells with linear, water soluble polyacrylamide-hydrazide chains followed by the stoichiometric addition of glyoxal as the cross linking agent (PAAH-G entrapment). In parallel, some cells were entrapped in calcium-alginate beads, as previously described. The capability of both immobilized cell systems to reduce monoterpenes was compared with freely suspended MENTHA cells. Entrapment by either alginate or PAAH-G did not impair cell vitality, as observed by fluorescein diacetate staining. Biotransformation of (-) menthone to (+) neomenthol by M-cells and of (+) pulegone to (+) isomenthone by P-cells indicated that the transformation efficiency of the cells entrapped in PAAH-G is as high as that of freely suspended cells. Moreover, the distribution of both precursor and product in the medium versus their content in the cells (or cells contained in gel-beads) showed that less monoterpenes were retained in cells entrapped in PAAH-G, as compared to the freely suspended cells. Thus prolonged incubation (e.g. 24 hr), which usually results in appreciable loss of monoterpenes from the chloroform extract of freely suspended-cells, caused considerably less loss from the PAAH-G entrapped-cells. In a preliminary test it was shown that PAAH-G entrapped cells were capable to perform three, consecutive, batch-type monoterpene biotransformations, without significant decrease of transformation capability. The capability to immobilize living plant cells within this synthetic chemically crosslinked gel system, combined with the favourable beads/ free-medium ratio of monoterpene distribution, point towards a potential development of a continuous biotransformation process carried out by plant-cells entrapped in this system. PMID- 17405002 TI - Clonal Multiplication of Pinellia ternata by Tissue Culture. AB - In M URASHIGE and S KOOG'S liquid medium supplemented with 0.25 ppm 2,4 dichlorophenoxyacetic acid(2,4-D) a tuber segment reached a fresh weight multiplication ratio of 200 when grown for 4 weeks at a rotation speed of 60 rev/min, at 25 degrees and in continuous light. The growth ability was maintained for 5 generations. Addition of 0.5 ppm 2,4-D and 1 ppm kinetin stimulated production of a maximum number of regenerated plantlets from the tissue mass segment. The regeneration ability was also estimated. It is theoretically possible to obtain more than 4 x 10 (23) plants from a single tuber in a year by this clonal propagation technique. PMID- 17405003 TI - Tertiary Phenolic Alkaloids from Chasmanthera dependens. AB - A phytochemical investigation of the tertiary phenolic alkaloidal fraction of the stems of CHASMANTHERA DEPENDENS H OCHST. (Menispermaceae) has yielded the tetrahydroprotoberberine type alkaloids govanine ( identical with(-) tetrahydropseudocolumbamine) [1] and coreximine [2]; the pavine type alkaloid bisnorargemonine [3] and the morphinandienone type alkaloid pallidine [4]. PMID- 17405004 TI - [Alkaloids from annonaceae.]. AB - From diverse parts of POLYALTHIA NITIDISSIMA, thirteen isoquinoline alkaloids have been isolated and identified. They belong to the structural types of benzylisoquinolines, aporphinoids, protoberberines and mainly bisbenzylisoquinolines. Four of these dimers, namely N,N'-dimethyllindoldhamine, isodaurisoline, 7-O-methyllindoldhamine and 7'-O-methyllindoldhamine, are new natural products. PMID- 17405005 TI - [Guattegaumerine, New Bisbenzylisoquinoline Alkaloid from Guatteria gaumeri.]. AB - The aqueous-alcoholic extract of the bark of GUATTERIA GAUMERI (Mexican Annonaceae) is used for the treatment of cholelithiasis and hypercholesterolemia. Therefore it seems very interesting to perform a phytochemical study of this drug in order to know its composition and to determine, if possible, all the active principles. We describe now the isolation of the main alkaloid named guattegaumerine. This new bisbenzylisoquinoline alkaloid has been assigned structure 1, on the basis of spectroscopic, chemical and chromatographic evidence. PMID- 17405007 TI - [An iso-Homoprotoberberin-Alkaloid from the Roots of Berberis actinacantha.]. AB - An alkaloid of the molecular formula C (21)H (19)NO (6) was isolated from the roots of berberis actinacantha. Substructures were derived from proton and carbon 13-NMR data. Single-crystal x-ray diffractometry identified the alkaloid as 15( R,S)-3,4-dimethoxy-11,12-methylenedioxy-8,14-dioxo-hexahydro- ISO homoprotoberberin. ORTEP plots, bond angles, assigned carbon-13 and proton NMR data are given. PMID- 17405008 TI - Chemical identification of alexandrian and tinnevelly senna. AB - Differentiation of CASSIA SENNA and C. ANGUSTIFOLIA is possible by chromatographic identification of the naphthalene glycosides. PMID- 17405009 TI - Steroidal Sapogenins in Dioscorea collettii. AB - Ten Steroidal compounds were isolated from the acid-treated rhizome of wild DIOSCOREA COLLETTII collected from Sichuan province, China. Five of them were identified as diosgenin ( 1), yamogenin ( 2), beta-sitosterol ( 3), Delta (3,5) deoxytigogenin ( 5) and isonarthogenin ( 9). The other four compounds have not been reported previously in the literature and their structures have been elucidated by spectroscopic data and chemical transformation. They are Delta (3,5)-deoxyneotigogenin ( 6), diosgenin palmitate ( 7), yamogenin palmitate ( 8) and yamogenin-beta-D-glucoside ( 10). It is of interest to note that three pairs of C (25) isomers were isolated: 1 and 2,5 and 6,7 and 8, apparently as artefacts due to the acid treatment. The structure of dihydroxysterol ( 4) is still to be determined. PMID- 17405010 TI - Alkaloids from Papaver triniifolium of Turkish Origin1. AB - The major alkaloids of PAPAVER TRINIIFOLIUM collected in two different parts of Anatolia have proved to be of the aporphine, morphinane and rhoeadine types. The two collections possessed different major alkaloids and the existence of chemical strains is indicated. In addition to the aporphine-(floripavidine), the morphinandienone-(amurine and salutaridine), benzylisoquinoline-(papaverine), proaporphine-(mecambrine) and tetrahydroprotoberberine-(sinactine, scoulerine and cheilanthifoline) type alkaloids have been isolated as minor alkaloids. PMID- 17405013 TI - Thalifaberine, thalifabine and huangshanine, three new dimeric aporphine benzylisoquinoline alkaloids. PMID- 17405014 TI - [not available]. PMID- 17405015 TI - Chemical Studies on Dangshen, the Root of Codonopsis pilosula. PMID- 17405017 TI - Alkaloids from Roots of Alstonia yunnanensis. PMID- 17405016 TI - A New Furanoflavone from Seeds of Pongamia glabra. PMID- 17405018 TI - Isolation of a New Furano-1,4-Naphthaquinone, Diodantunezone from Lanthana achyranthifolia1. PMID- 17405019 TI - Valepotriates in Valeriana thalictroides. PMID- 17405021 TI - [New Cardiac Glycosides from Digitalis lanata.]. AB - From the leaves of DIGITALIS LANATA E HRH. three as yet unknown cardenolides were isolated and structurally elucidated as Digoxigenin-3-O-beta-D-digitoxosido-beta D-digitoxosido-beta-D-glucomethylosid, Digoxigenin-3-O-beta-D-digitoxosido-beta-D glucomethylosid and Diginatigenin-3-O-beta-D-digitalosid by TLC and mainly spectroscopic methods. PMID- 17405022 TI - The Role of 9- and/or 10-oxygenated Derivatives of Geraniol, Geranial, Nerol, and Neral in the Biosynthesis of Loganin and Ajmalicine. AB - A number of deuterium labelled 9- and/or 10-oxygenated derivatives of geraniol, geranial, nerol, and neral were used as substrates in feeding experiments with a cell suspension culture of CATHARANTHUS ROSEUS. The 9- and 10-oxo derivatives of geraniol, geranial, nerol, and neral as well as 9,10-dihydroxygeraniol, 9-oxo-10 hydroxygeraniol, and 9,9,10,10-tetraethoxygeranial were found to be good precursors of the indole alkaloid ajmalicine. The compounds 9-hydroxygeraniol, 9 hydroxynerol, 9,10-dihydroxynerol, 9-oxo-10-hydroxynerol, 9-hydroxy-10-oxonerol, 9-hydroxy-10-oxogeraniol, 9,9,10,10-tetraethoxynerol, and 9,9,10,10 tetraethoxygeraniol were relatively poor precursors. Competitive feeding experiments, employing mixtures of unlabelled citral and labelled 9-oxogeranial, 10-oxonerol, 10-oxoneral, or 9,10-dihydroxygeraniol were also carried out. The incorporation of 9,10-dihydroxygeraniol into ajmalicine was not significantly affected by the presence of citral, while the incorporations of the other labelled substrates were greatly reduced. PMID- 17405023 TI - Alkaloids of Aconitum barbatum. AB - Nine diterpenoid alkaloids were isolated from the roots of ACONITUM BARBATUM VAR. PUBERULUM. Five of them are known: ranaconitine ( 1) [1], lappaconitine ( 2) [2], septentriodine ( 3) [3], septentrionine ( 4) [3], and lycaconitine ( 5) [4]. The other four are new: puberanine ( 6), puberanidine ( 7), puberaconitine ( 8) and puberaconitidine ( 9). PMID- 17405024 TI - [Components of Root Bark of Morus australis.]. AB - In addition to mulberrofuran D, a new 2-arylbenzofuran derivative, six flavone derivatives, morusin, oxydihydromorusin, cyclomorusin, kuwanon C, G and H, were isolated from extracts of root bark of MORUS AUSTRALIS P OIR. The structure of mulberrofuran D was shown to be 5 on the basis of spectral and chemical data. Mulberrofuran D is the first example of a geranylated and prenylated 2 arylbenzofuran derivative found in nature. PMID- 17405025 TI - Novel Structures of two Chromone Alkaloids from Root-Bark of Schumanniophyton magnificum. AB - The constitutional formulae of two new chromone alkaloids, schumannificine 2 and N-methylschumannificine 3 isolated from the root-bark of SCHUMANNIOPHYTON MAGNIFICUM, H ARMS, have been shown to be linear tetracyclic compounds with ring D being piperidine in nature, on the basis of the chemical evidence and spectral analyses. PMID- 17405030 TI - Effect of Culture Conditions on DOPA Accumulation in a Callus Culture of Stizolobium hassjoo. AB - Effects of passage of subculture and several culture conditions, nutritional, hormonal and light conditions on 3,4-dihydroxyphenylalanine (DOPA) content were examined in callus of STIZOLOBIUM HASSJOO P IPER and T RACY. The favourable conditions for DOPA accumulation were found to be suboptimal conditions for growth, i.e., low concentration of phosphate, low concentration of 2,4-D, high concentration of kinetin. Light promoted DOPA accumulation. The best condition for DOPA accumulation was discussed in relation to protein synthesis. PMID- 17405031 TI - Isolation of New Alkaloids from Catharanthus roseus. PMID- 17405032 TI - Isolation of Moenjodaramine from Buxus papilosa. PMID- 17405033 TI - A Sesquiterpenoid Naphthol from Kydia calycina. PMID- 17405035 TI - Partial Purification and Properties of S-Adenosylmethionine: (R), (S) Norlaudanosoline-6-O-Methyltransferase from Argemone platyceras Cell Cultures. AB - A new enzyme, S-adenosylmethionine: (R), (S)-norlaudanosoline-6-O methyltransferase, was isolated from the soluble protein extract of A. PLATYCERAS cell cultures and purified approximately 80-fold. This enzyme catalyses the formation of 6-O-methylnorlaudanosoline, and, to a minor extent, 7-O methylnorlaudanosoline from SAM and (S), as well as (R), norlaudanosoline. The apparent molcular weight of the enzyme is 47000 Dalton. The pH-optimum of the enzyme is 7.5, the temperature optimum, 35 degrees C. Apparent K (M) values for (S) and (R)-norlaudanosoline were 0.2 mM, and for SAM, 0.05 mM. The transferase shows high substrate specificity for tetrahydrobenzylisoquinoline alkaloids. Simple orthophenols, like phenylpropane derivatives, coumarins or flavonoids, are not accepted as substrates. The enzyme is widely distributed in benzylisoquinoline-containing plant cell cultures and is present in differentiated plants like PAPAVER SOMNIFERUM. PMID- 17405037 TI - New Acylated Cyanogenic Diglycosides from Fruits of Anthemis cairica. AB - Three new cyanogenic diglycosides of the mandelonitrile series have been isolated from fruits of ANTHEMIS CAIRICA (Compositae) and their structures identified by degradation and spectral methods mainly (1)H-NMR, (13)C-NMR, FAB-MS. Minor compounds are 2-beta-primeverosyloxy-2-phenyl-2S-acetonitrile (epilucumin) and its 4''-p(beta-D-glucopyranosyloxyl-(E)cinnamate; the main compound is the 4'' p(beta-primeverosyloxy)-(E)cinnamate of epilucumin. PMID- 17405038 TI - [Analysis of linden flower mucilage.]. AB - Linden flowers are known to contain considerable amounts of mucilage. Quantitative determinations of these water soluble polysaccharides were carried out by viscosimetric measurements. It was shown that the mucilage after total hydrolysis in trifluoracetic acid was composed of D-galactose, L-arabinose and L rhamnose and minor amounts of D-glucose, D-mannose and D-xylose. The high percentage of uronic acid (52%) was due to the presence of both, D-galacturonic acid and D-glucuronic acid. Fractionation of the mucilage polysaccharides on DEAE cellulose resulted in a separation of five distinct polysaccharide fractions, which showed different sugar compositions and uronic acid content. The homogeneity of the polysaccharide fractions 1-5 was further examined by high voltage electrophoresis and analytical ultracentrifugation. PMID- 17405039 TI - [Chemical Constituents of the Convolvulaceae-Resins IV1.]. AB - From the roots and upper parts of the Convolvulaceae IPOMOEA QUAMOCLIT, I. LACUNOSA, I. PANDURATA and CONVOLVULUS AL-SIRENSIS by ether-, petrolether and ethanol extraction respectively a resin material could be isolated, which yielded after alcaline hydrolysis glycosidic acids with jalapinolic acid as the common hydroxy fatty acid and the sugars rhamnose, glucose and xylose in different molar ratios. PMID- 17405041 TI - 1HNMR Chemical Shift Values for Aromatic Protons in 2,3,9,10- and 2,3,10,11 tetrasubstituted Tetrahydroprotoberberine Alkaloids. AB - The (1)HNMR (CDCl (3)) data on 28 tetrahydroprotoberberine alkaloids have shown that it is possible, from the chemical shift values observed for the aromatic protons, not only to determine the substitution pattern but also the type of substituents carried on the two sites in rings A and D within this class of alkaloids. PMID- 17405042 TI - New Oleanene-type Triterpenes from Abrus precatorius and X-ray Crystal Structure of Abrusgenic Acid-Methanol 1:1 Solvate. AB - From ABRUS PRECATORIUS Abl. triterpenoids abruslactone, methyl abrusgenate and abrusgenic acid were isolated and elucidated by spectroscopic and X-ray techniques. PMID- 17405043 TI - [Tannin Producing Dimeric Proanthocyanidins in Juniperus communis.]. AB - From bark extracts of JUNIPERUS COMMUNIS L. (Cupressaceae) some dimeric proanthocyanidins have been isolated and their structures elucidated. They belong to the procyanidin typs B1, B2, B3 and B4, what means that there is a C4 to C8' linkage. PMID- 17405044 TI - Biosynthesis of Sterols and Triterpenoids in Tissue Cultures of Cucurbita maxima. AB - The biosynthesis of sterols and triterpenoids in CUCURBITA MAXIMA was studied by analysis of unsaponifiable fraction of tissues from different development stages of the plant (seeds, seedlings, adult plant and tissue culture) and by feeding germinating seeds and tissue cultures with [2- (14)C]-acetate. Synthesis of cucurbitacins does not occur in callus tissues of CUCURBITA MAXIMA, whereas a wide variety of 4,4-dimethylsterols present in these tissues testifies of a high level of squaleneoxide cyclase activity in growing callus. The peculiarity of Cucurbitaceae among the higher plants is also discussed comparing the side chain biosynthesis of sterols in CUCURBITA MAXIMA to that operating in other higher plants. PMID- 17405045 TI - Isolation and structure of a rarely occurring cyanidanol glycoside from cortex betulae. AB - Cortex Betulae contains (+)-catechin-7-beta-D-xylopyranoside. Except for its occurrence in elm bark, this biologically active substance is so far unknown as a natural product. PMID- 17405046 TI - Structure of hydroxymuscopyridine a and hydroxymuscopyridine B, two new constituents of musk. AB - The structural elucidation of hydroxymuscopyridine A ( 3) and hydroxymuscopyridine B ( 5), two new constituents isolated from musk, MOSCHUS MOSCHIFERUS, is reported. PMID- 17405049 TI - [RP-8 Auxiliary Phase for the Accumulation of Valepotriates from Cell-Suspension Culture of Valeriana wallichii.]. PMID- 17405050 TI - Editorial. PMID- 17405051 TI - A Highly Specific O-Methyltransferase for Nororientaline Synthesis Isolated from Argemone platyceras Cell Cultures. AB - A highly specific new enzyme, S-adenosyl-L-methionine: (6-O-methyl norlaudanosoline)-5'-O-methyltransferase which catalyses the formation of nor orientaline from 6-O-methyl-norlaudanosoline and SAM was discovered, partially purified, and characterized. ARGEMONE PLATYCERAS cell suspension cultures served as enzyme source. PMID- 17405054 TI - Studies on Iridoid-related Compounds III: Gentiopicral, the Aglucone of Gentiopicroside. AB - Enzymatic hydrolysis of gentiopicroside ( 1) provided the new aglucone, gentiopicral ( 2), the structure of which was firmly elucidated by means of spectroscopic methods. PMID- 17405053 TI - HPLC Separation and Quantitative Determination of Biflavones in Leaves from Ginkgo biloba. AB - An HPLC method for separation and quantitative determination of biflavones in crude leaf extracts from GINKGO BILOBA L. is described. A system using a Lichrosorb(R)-Diol column and the ternary elution system: hexane-chloroform tetrahydrofuran, was suitable for separation of sciadopitysin, ginkgetin, isoginkgetin and bilobetin. The biflavones were detected at 330 nm; their calibration curves were established and their response factors were calculated using acacetin as internal standard. This method was used for the quantitative determination of the biflavones in crude leaf extracts; the seasonal variations of the biflavone content were studied and showed a higher amount in autumn leaves. PMID- 17405055 TI - Gentiogenal, a conversion product of gentiopicrin (gentiopicroside)*. AB - The structure of gentiogenal, (+/-)5-formyl-6-methyl-3,4-dihydro-1H,6H-pyrano [3,4-c-]-pyran-1-one, a conversion product of the aglucone of gentiopicrin (gentiopicroside), isolated from BLACKSTONIA PERFOLIATA (Gentianaceae), was elucidated by (1)H- and (13)C NMR spectroscopic, mass spectrometric and X-ray diffraction methods. In a TLC bioassay gentiogenal showed fungitoxicity towards PENICILLIUM EXPANSUM. PMID- 17405056 TI - Mono- and Diterpenes from Cell Cultures of Thuja occidentalis. AB - Cell cultures of THUJA OCCIDENTIALIS L. were found to biosynthesize various mono- and diterpenes when grown on B5-medium. The identification of the constituents was achieved mainly by capillary GLC-MS using fused silica columns and E.I.-mass spectrometry. Monoterpenes of the menthane type were only isolated from the culture medium whereas diterpenes were found in the cell extracts. Thujaplicin derivatives, monoterpenes of an irregular type, were detected in the medium as well as in the cells. Major differences were found between the terpene composition of the cell culture extracts and those from THUJA leaves. The cell cultures accumulated some compounds which are presently unknown as constituents of THUJA plants. On the other hand, the cultures were evidently unable to synthesize the thujone type of monoterpenes. PMID- 17405058 TI - [Helenalin- and 11,13-Dihydrohelenalinester from Flowers of Arnica montana.]. AB - From the flowers of ARNICA MONTANA L. 13 helenanolides were isolated and identified. They are shown to be 11,13-dihydrohelenalin ( 1), helenalin ( 2) and their ester derivatives 3-13 (see figure). The natural occurence of 6-O isobutyryl-, 6-O-tigloyl-, 6-O-isovaleryl- and 6-O-(2-methyl)-butyrylhelenalin (compounds 9, 11, 12, 13) is reported for the first time. The qualitative sesquiterpene lactone composition in flowers of A. MONTANA from different regions was found to be variable. PMID- 17405059 TI - Dichomine, a novel type of iboga alkaloid*,**. AB - From the leaves of TABERNAEMONTANA DICHOTOMA and the leaves and twigs of T. EGLANDULOSA a new alkaloid was isolated which was assigned structure 1 on the basis of spectroscopic and chemical evidence. PMID- 17405060 TI - Essential Oil of Chrysanthemum indicum. AB - The steam distilled oils both from flowers and leaves, as well as from total aerial parts of CHRYSANTHEMUM INDICUM L. (small-flowered form), were obtained and analyzed by GC and GC/MS. Their main components are borneol, chrysanthenone and bornyl acetate. PMID- 17405061 TI - New 6-Aryl-2-pyrones from Gentiana pedicellata*. AB - Three new 6-aryl-2-pyrones, viz. pedicellin ( 1), pedicellanin ( 2), and pedicellatin ( 3), have been isolated from the whole plant of GENTIANA PEDICELLATA W ALL., collected when in flower. The compounds have been characterized on the basis of chemical transformation and comprehensive spectral evidence. This is the first report of occurrence of 6-aryl-2-pyrones in a member of the family Gentianaceae. The other secondary metabolites, e.g. triterpene, secoiridoid, and degraded carotenoid, present in this species are typical of the GENTIANA. 1 and 2 produced significant adaptogenic and anti-stress effects in animal models thereby justifying the use of the title species in popular medicine. PMID- 17405062 TI - [On the Constituents of Isertia hypoleuca.]. AB - Dihydroquinamine and 3-epi-dihydroquinamine were isolated and identified from ISERTIA HYPOLEUCA (Schultes, Rauffauf and Soejarto 24041). The structure of hydroxydihydrocinchonamine, a reduction product from dihydroquinamine with LiAlH (4), was established. The (13)C magnetic resonance spectra of the three alkaloids have been determined. PMID- 17405063 TI - 8-Epidesacylcynaropicrin from Crepis capillaris. AB - The aerial parts of CREPIS CAPILLARIS afforded a new guaianolide. The structure of this compound has been determined as 8-epidesacylcynaropicrin. PMID- 17405064 TI - Constituents of Essential Oils from Majorana syriaca, Coridothymus capitatus and Satureja thymbra*. AB - The essential oils of MAJORANA SYRIACA, CORIDOTHYMUS CAPITATUS and SATUREJA THYMBRA plants growing wild in Israel were investigated by means of GLC and GLC MS. 18 components were identified, the main ones being thymol, carvacrol, gamma terpinene and p-cymene. PMID- 17405065 TI - [Detection of Cannabinoids in Phelipaea ramosa, a Parasite of Cannabis sativa.]. AB - Cannabinoids have been characterized in PHELIPAEA RAMOSA L. which lives as a parasite on Hemp ( CANNABIS SATIVA L.). This confirms that these substances are not exclusively found in glandular trichomes of chlorophyllian organs of the CANNABIS. PMID- 17405067 TI - A Note on "Isolation and Pharmacological Action of Heliotrine, the Major Alkaloid of Heliotropium indicum Seeds". PMID- 17405069 TI - [Guidelines for cardiopulmonary resuscitation in preterm and term neonates]. PMID- 17405070 TI - [Sweat testing practices in German cystic fibrosis centres]. AB - BACKGROUND: Surveys in USA and UK have shown inconsistencies between practice of sweat testing and guidelines of the National Committee for Clinical Laboratory Standards (NCCLS), which could increase the risk of false-positive and -negative results. To determine the current practice of sweat testing in German cystic fibrosis (CF)-centres a survey was performed. METHOD: A questionnaire was mailed to all 123 German CF-centres that join the German CF-register. The results were compared with the NCCLS-guidelines. RESULTS: 88 out of 113 responding centres perform 30-1100 (median 200) sweat tests by pilocarpine iontophoresis per year. The recommended sweat sample time of 20-30 min was accepted by 33% of all centres. Minimum acceptable sweat rates were quoted between 100 and 7500 (median 1000) mg/m (2)/min differing from the recommended 1000 mg/m (2)/min. Osmolality was measured in 3%, sodium in 24% and conductivity in 53% of the centres. Chloride, the only accepted diagnostic measurement by the NCCLS, was investigated by only 58% of all centres. The recommended chloride CF-reference value of >60 mmol/l was used in 78% of centres that perform chloride testing. CONCLUSIONS: Major areas of inconsistencies between sweat testing practice in German CF centres and NCCLS-guidelines are sweat collection, analyzing method and reference values. Chloride, the only accepted diagnostic measurement by the NCCLS, was investigated only by 58% of all centres. PMID- 17405071 TI - Renal complications associated with human parvovirus B19 infection in early childhood. AB - A previously healthy two-year-old girl presented with proteinuria and macroscopic haematuria. Laboratory findings included haemolytic anaemia with thrombocytopenia. Interestingly, continuing reticulocytopenia was noted. Therefore an acute parvovirus B19 infection was suspected, which could be confirmed by serological and molecularbiological evidence. This case report underlines renal complications of parvovirus B19 infection in early childhood including haemolytic-uraemic syndrome (HUS)-like episodes, and potential pathogenetic mechanisms are discussed. PMID- 17405072 TI - [Exploration of cognitive processes in children and adolescents by means of visual event related potentials]. AB - BACKGROUND: Viewing pictures activates the human brain in a complex manner. Stimulated brain areas can be identified by electrophysiological methods even in children. SUBJECTS AND METHOD: Event related potentials (ERP) were recorded with EEG-surface electrodes in children and adolescents who were shown pictures of increasing complexity. Pictures were categorized into morphologic features such as brightness, color and pattern as well as semantic content. RESULTS: Presentation of comics and unstructured pictures, similar in color and brightness, revealed ERP with nearly identical early components (P 100). However, substantial differences were found in frontocentral brain areas. A negative wave at approximately 400 ms was recorded only when comics were shown. CONCLUSIONS: Semantic scenes provoked significant different ERP, which indicate an early involvement of complex neuronal networks. These results could be applied in the research of pediatric cognitive disturbances. PMID- 17405073 TI - [Asperger syndrome - a too late diagnosed disorder?]. AB - Asperger syndrome is a disorder within the autistic spectrum, which was first described by Hans Asperger in 1944. It belongs to the group of pervasive developmental disorders and is particularly characterized by qualitative impairments of social interaction and communication as well as distinct special interests and stereotyped patterns of behaviour. We present a patient, showing the typical behavioural symptoms of the Asperger syndrome, which were first diagnosed at the age of sixteen. PMID- 17405074 TI - Diencephalic syndrome: a frequently delayed diagnosis in failure to thrive. AB - BACKGROUND: Diencephalic syndrome (DS) is a rare cause of failure to thrive in early childhood. It is associated with neoplastic lesions of the hypothalamic optic chiasmatic region. Treatment options consist of surgical resection, radiation therapy (RT) and chemotherapy. We describe the clinical course of two children suffering from diencephalic syndrome due to unresectable hypothalamic gliomas and emphasize the importance of chemotherapy as a first-line treatment. PATIENTS AND METHODS: We report about two children, at the age of 21 months and 13 months at diagnosis, who presented with severe dystrophy at 12 months and 6 months respectively. Imaging of the brain showed a suprasellar mass, identified histologically as low grade pilocytic astrocytoma. Both patients were treated with chemotherapy which induced tumor regression and stable disease. RESULTS: The two children gradually gained weight and improved remaining in stable remission. CONCLUSIONS: Diencephalic syndrome caused by a hypothalamic/chiasmatic astrocytoma is a rare cause of failure to thrive in children so that diagnosis is frequently delayed. It should be considered as differential diagnosis in any child with dystrophy despite adequate caloric intake. Since most of these tumors in that specific anatomic site are regarded to be unresectable, chemotherapy including carboplatin and vincristine may reveal clinical improvement in these patients. PMID- 17405075 TI - Cerebral sinus occlusion in a boy presenting with asparaginase-induced hypertriglyceridemia. AB - Cerebral sinus thrombosis is a rare but severe complication during treatment for acute lymphoblastic leukaemia (ALL). It mostly has been reported during treatment with asparaginase and dexamethasone. Hypertriglyceridemia has - albeit very rarely - also been associated with asparaginase therapy. The combination of cerebral sinus thrombosis and hypertriglyceridemia however, has not yet been reported. Here we describe a 15-year-old boy who presented with clinical symptoms and radiologic findings of a cerebral sinus thrombosis. In addition, a life threatening hypertriglyceridemia was present. The complication was successfully treated by anticoagulation with low molecular weight heparin and the lipid regulator bezafibrate. PMID- 17405077 TI - [The brave new world of prevention? On the prerequisites and scope of public health genetics]. AB - Recent developments in molecular genetics and genomics have not only provided new insights into the biological foundation of life but have also brought about significant changes in the ways health and disease are understood. Nowadays, understanding how the human genome interacts with health-related behaviour and diverse environments is recognised to be the key for biomedical innovation. Even though advances in genetics and genomics have unveiled a whole new complexity of the genetic underpinnings of health and disease and clearly show the need for further investigation, studies of health-related genetic traits already give rise to novel approaches to prevention. As a relatively new fieldPublic Health Genetics is striving for the integration of genetic knowledge into strategies of prevention and for the translation of genomics into novel tools for public health. Translational research in this field is in its very early stages. The rise of public health genetics together with the relative predictive power of genetic information urges us to address the fundamental social, economical and ethical issues of public health genetics. This study analyses the scientific venture points and the larger framework of the implementation of strategies of public health genetics. The applicability of predictive and preventive measures of public health genetics are scrutinised with regard to their effect on both public and individual health. A major focus is on the economical, social and ethical challenges arising from any implementation of public health genetics. The guiding principle of our analysis is the (improvement of) social achievability of health as a measure for successful innovation of health care and prevention. PMID- 17405078 TI - [Work-site health promotion in Germany. Results of the IAB--establishment panel 2002 and 2004]. AB - According to the answers of employers to the representative IAB establishment panel 2004, one-fifth of the companies in Germany voluntarily carried out or financially supported measures for the protection or promotion of the health and well-being of their work force. The proportion of health-promoting companies was above average in all East German federal states as well as in Bavaria, in Saarland and in Lower Saxony. North Rhine-Westphalia was precisely average. In East Germany, almost one-fourth and in West Germany just under one-fifth of all companies surveyed carry out health-promoting measures.Work-site health promotion varies considerably depending on the corresponding federal states, industrial branches and company sizes. Work-site health promotion has ,up to now, been concentrated in big companies and groups. An under-representation of work-site health promotion is observed above all in small and very small companies and particularly in the catering trade. Work-site health promotion was principally determined by analyses of sickness leaves and surveys on health protection in the work place which were mentioned in about 9% and 8% of the companies interviewed. 6% of the companies provided courses on health-relevant ways of behaviour. About 4% of the companies maintain health circles and 5% realised other health promoting measures. Further correlation analyses reveal that in companies with a workers council/staff council, work-site health promotion was significantly much more common. This correlation is especially strong in small and medium-size companies. The longitudinal analysis of the IAB establishment panel 2002 and 2004 reveals much dynamism in work-site health promotion. On the one hand, half of the companies involved in work-site health promotion in 2002 had stopped their commitment to work-site health promotion by 2004. Sustainability remains one of the biggest challenges in work-site health promotion. On the other hand, a bit more than one tenth of the repeatedly interviewed companies started health promoting activities in 2004. According to the longitudinal data set, altogether 29% of the companies were reached by work-site health promotion measures (for the years 2002 and 2004). PMID- 17405079 TI - [Community-based health promotion--a challenge for the evaluation]. AB - PURPOSE: Community-based health promotion (CBHP) aims at mobilising citizens for health-related issues in their environment, and at implementing health-promoting projects on the community level. Whereas recent political decisions support this approach, scientific studies dealing with theories and consequences of CBHP are scarce in Germany. Evaluation of CBHP could help identify (in)effective factors and elements of community programmes and thus improve future planning. In Germany, however, there is a deficit in systematic concepts and recommendations for the evaluation of CBHP. This work outlines basic ideas and core principles of CBHP and deduces implications for the assessment of health-promoting community projects. METHODS: Based on different international models and studies and on discussions with health promotion professionals, we developed a framework for the evaluation of CBHP. RESULTS: The proposed framework includes a guideline for CBHP programme planning. Its strategic and operational criteria can serve as a basis for a strategy evaluation. In terms of process evaluation, indicators for the dimensions (1) programme implementation and service delivery, (2) capacity building, and (3) reach of and acceptability in the target group were developed. In addition, we present different areas of OUTCOME EVALUATION; it is advisable to distinguish between measurement on the individual and on the community level. The framework further proposes strategies for the evaluation of the core principles empowerment and participation. CONCLUSION: The presented framework can serve as a basis for the development of flexible and individual instruments for the evaluation of CBHP, which should not ignore the perspective of the citizens, or complex aspects like changes on the community level. Some aspects, e.g., the potential evaluation of further targets of CBHP (improvement of quality of life, reduction of social and health inequalities), the responsibility of evaluation or the effects of financial constraints, are important questions to be addressed in the future. PMID- 17405080 TI - [Self-determined but with professional leadership? On the effectiveness and definition of self-help groups]. AB - PURPOSE: Because the effectiveness of self-help groups (SHGs) has not been proven yet and also since multi-focus reviews on SHGs are rare, the present article offers an overview of the current state of research. Possible reasons for the conflicting results in the literature are also investigated. METHODS: An extensive literature search was done to review studies which used longitudinal design and control groups to measure the effectiveness of SHGs and so-called "support groups". RESULTS: Positive effects in comparison to the control groups were proven in four out of seven of the studies. In the remaining three studies the outcomes between the groups were the same. CONCLUSION: Considering the naturalistic designs of the studies, these results clearly indicate that SHGs are indeed effective. An insufficient consideration of the heterogeneity of the groups is held responsible for the difficulties associated with previous research on the topic, especially as far as the differentiation between SHGs and professionally led groups is concerned. The definition excludes professional leadership but, in fact, professional involvement in SHGs up to their leadership is the rule. The following suggestions have been derived from this literature analysis: uniform classifications for SHGs and professionally-led support groups should be established in order to make it possible to provide a clear distinction between these types of groups as well as to indicate the extent of professional involvement in them. A plea is made for a generally stronger consideration of organisational differences in the research of SHGs. PMID- 17405081 TI - [Patient and public involvement in health care--a comparison between recent english and german policies]. AB - AIMS: Involving patients and citizens in health care and policy has become a regular part of health care reform programmes in highly developed societies. Systematic knowledge about particular initiatives and their implementation is mostly lacking as are comparative approaches. In this article recent attempts to promote patient and public involvement in England and Germany are analysed based on available evidence. These two countries represent different types of health care systems and their governance but share a similar history of paternalistic styles of health policy making and health care provision. RESULTS: This study shows various recent policy-initiatives in both countries. In total, English policies cover a broader spectrum and are better institutionalized. This applies especially in the field of patient information, patient surveys and for collective participation on various policy levels. The most remarkable German policies can be found in the fields of patient advice and support of self-help groups. Recently institutionalized mechanisms of collective involvement have to be considered as initial steps; preliminary experience shows the need for further development. CONCLUSIONS: Experiences in both countries demonstrate that a profound cultural change towards more patient and public involvement in health will have to be realised with a long-term perspective and will have to learn from ongoing practice. Sufficient support, adequate information and transparency will be essential to get and keep the lay public involved. PMID- 17405082 TI - [Dental health in German and Turkish school children--a 10-year comparison]. AB - The purpose of this cross-sectional study was to assess the changes in dental health in 12- and 15-year-old Turkish immigrants and German students who were included in a school-based caries-preventive programme for ten years (1993-2003). In 1993 352 and 402 12- and 15-year-olds and in 2003 352 and 402 12- and 15-year olds were investigated, respectively. 23.6% of the 12-year-olds and 21.5% of the 15-year-olds were of Turkish origin. The clinical examination was performed with support of artificial light at school. Caries was diagnosed according to the WHO standard using the DMFT Index for permanent dentition. Within the ten-year period the highest increment of caries-free dentitions was found in 12- and 15-year old Germans attending grammar schools with 41,5% and 27% and secondary modern schools with 22,3% and 17%, respectively. The proportion of caries-free Turkish students attending secondary modern schools, only increased imperceptible in both age groups (1.2%). A significant caries decline of 1.5 DMFT was observed in 12-year old Germans at grammar schools, only. In 15-year olds caries declined in German students at secondary modern schools (2.5 DMFT), and at Grammar schools (2.2 DMFT), significantly, however caries experience in Turkish students remained nearly unchanged (0.3 DMFT). A strong polarisation of dental caries was found in 2003. Among the 12-year-olds 23% of those at secondary modern school accumulated 70% of the total caries experience (>3 DMFT) and at grammar school 9% of students accumulated 76% of the whole amount of caries (>1 DMFT). Among the 15-year-olds 73% of the total caries experience (>3 DMFT) was concentrated in 32% of the students at secondary modern schools, while 25% of the students at grammar schools exhibited 84% of the whole amount of caries (>1 DMFT). Intended concepts in caries prevention in the future should aim to improve dental health especially in Turkish students at secondary modern schools while taking social inequality of this risk group into account. PMID- 17405083 TI - Phthalides in Roots of Opopanax chironium. AB - In petroleum ether and diethyl ether extracts from roots of Opopanax chironium (L.) Koch five phthalides were identified: Z-butylidenephthalide, butylphthalide, cnidilide, Z-ligustilide and senkyunolide. The compounds were analyzed by TLC, GLC and GC-MS. PMID- 17405084 TI - Biotransformation of monoterpenes by Mentha cell lines. AB - Previous results indicated that all tested cell suspension lines derived from various Mentha chemotypes were capable to biotransform (-) menthone to (+) neomentol but only some of these lines converted (+)-pulegone to (+)-isomenthone. In order to quest whether only the natural secondary metabolites or also other compounds with similarities to pulegone are biohydrogenated by Mentha cell suspensions, we incubated such suspensions with 5 unsaturated alpha-beta; ketones. No conversion was detected when mesityl oxide, trans-6-tert. butyl pulegone or 3-isopropylidine-9-methyl-decalone-2 were incubated with Mentha cells, while saturation of the alpha-beta double bond of 2-isopropylidine cyclohexanone and of trans-6-methyl pulegone was observed in suspensions of those cell lines which are capable of pulegone transformation. Suspensions of Mentha cell lines which were incapable to hydrogenate pulegone did not biotransform the two latter pulegone analogues. PMID- 17405085 TI - Growth and alkaloid production in suspension cultures of Hyoscyamus muticus as influenced by various cultural parameters. AB - Suspension cultures of Hyoscyamus muticus L. were raised in RT medium supplemented with NAA 10 (-5)M. Effects of pH, vitamins, sucrose, sodium and potassium succinate, chemical precursors and phytohormones on growth, morphogenesis and total tropane alkaloid production have been examined. Of these, low pH, sodium succinate, precursors and IAA enhanced alkaloid production. Alkaloid synthesis was higher during stationary phase and was found to be independent of morphogenesis. PMID- 17405087 TI - Two bitter biphenyl glucosides from Swertia japonica. AB - From the whole plants of Swertia japonica Makino a mixture of two new bitter biphenyl glucosides biphenosides A and B, was isolated along with three known secoiridoid glucosides by the application of droplet countercurrent chromatography (DCCC). The structure of the biphenyl glucoside was established by chemical degradation and spectroscopic methods. PMID- 17405088 TI - Constituents of the Chinese crude drug "sang-bai-pi" (Morus root bark). AB - From the benzene extract of the Chinese crude drug "Sang-Bai-Pi" (Japanese name Sohakuhi), the root bark of Morus sp. (Moraceae), a novel isoprene substituted flavanone, named sanggenon A, was isolated whose structure was shown to be I on the basis of spectral data. A known isoprene substituted flavone derivative, morusin (II), was also obtained from the extract. PMID- 17405089 TI - Pseudolaric acids from Pseudolarix kaempferi. AB - Three novel diterpenoids, pseudolaric acids A, B and C, were isolated from the root bark of Pseudolarix kaempferi Gorden used as an antifungal agent in folk medicine in China. The structures of pseudolaric acids B and C were assigned as 1 and 6 by spectral and chemical evidence. Pseudolaric acids A, B and C possess antifungal activity and pseudolaric acid B possesses an antifertility effect. PMID- 17405090 TI - Structures of Crassicaulisine and Crassicaulidine, two New Diterpenoid Alkaloids. AB - From the roots of Aconitum crassicaule, two new diterpenoid alkaloids, crassicaulisine and crassicaulidine, were isolated. The structure of crassicaulisine, C (24)H (39)O (7)N, has been elucidated as 3 on the basis of chemical and spectral data, while the structure of crassicaulidine, C (24)H (39)O (8)N, has been shown as 4 on the basis of chemical data and spectral comparison with crassicaulisine. PMID- 17405091 TI - Quantitation of Principal Bufadienolides in Different Cytotypes of Urginea indica. AB - Roots of URGINEA INDICA K UNTH, (Liliaceae) Indian squill, contain high amounts of proscillardin A and scillaren A. The range of variation in proscillaridin A and scillaren A contents in bulbs is 0.004-0.26% in diploids, 0.04-0.07% in triploids and 0.02-0.45% in tetraploids. The relative proportion of proscillaridin A and scillaren A also varies in different cytotypes. A tetraploid cytotype, representing a high yielding strain which can be exploited commercially for better yield of cardiac glycosides, has been obtained from Pune. PMID- 17405092 TI - New Flavanones from Prosopis juliflora Roots. AB - From the benzene and ethyl acetate soluble fractions of an ethanolic extract of roots of PROSOPIS JULIFLORA two new flavanone glycosides have been isolated and characterised as 3', 4'-dihydroxy 5-methoxy 6-methyl flavanone 7-0-beta-D glucopyranoside (I) and 7,4'-dimethoxy 6,8-dimethyl flavanone 5-0-beta-D galactopyranoside (II). PMID- 17405093 TI - Constituents of Essential Oil of Artemisia arborescens. AB - By combination of capillary GLC and mass spectrometry it was possible to analyze the composition of the essential oil obtained by steam distillation from leaves and flowers of ARTEMISIA ARBORESCENS L. growing in Sassari (Osilo) in Sardinia. This oil is coloured blue by the presence of relatively high amounts (11.32%) of chamazulene, a substance with anti-inflammatory properties. The oil, after separation in fractions on a silica gel column, was investigated by means of capillary GLC and capillary GLC-MS. Thujone, camphor and chamazulene account for about 75% of the oil. Almost a hundred minor components were found by capillary GLC and 44 were identified. PMID- 17405094 TI - A New Phenolic Glucoside, Curculigoside from Rhizomes of Curculigo orchioides. AB - A new phenolic glucoside, named curculigoside (I), was isolated from the rhizomes of CURCULIGO ORCHIOIDES G AERTN. and its structure was elucidated as 5-hydroxy-2 O-beta-D-glucopyranosylbenzyl 2,6-dimetoxybenzoate. PMID- 17405095 TI - [Nemoroside and nemorososide, new iridoid glucosides from Penstemon nemorosus]. AB - From leaves of Penstemon nemorosus Trautv. two new esteriridoids have been isolated. Iridoidic moieties are catapol and ajugol, both esterified with 8 hydroxy-2,6-dimethyl-2,6-octadienoic acid. The structure was established by spectroscopic methods and comparison with known compounds. PMID- 17405096 TI - [Composition of the essential oil from beta-Asarone free calamus.]. AB - The study of the essential oil extracted from american diploid calamus rhizomes proved that this variety does not contain the cancerogenic cis-isoasarone (beta asarone) nor other phenylpropane derivatives which are believed to be toxic. Typical compounds of the examined oil are sesquiterpeneketones, e. g. shyobunones and acorones which pharmaceutically used calamus oils also contain. We believe the isoasarone-free calamus rhizome to be an alternative to the triploid calamus variety often used in todays phytotherapy. PMID- 17405097 TI - [Extraction of thermolabile compounds with supercritical gases.]. AB - The thermolabile acoragermacrone and other sesquiterpenes could be extracted with supercritical carbon dioxide from beta-asarone free calamus rhizomes ( Acorus calamus L. var. americanus Wulff. A usual steam distillation lets this substance partly decompose into shyobunone. By using high pressure extraction with fractionated separation a higher yield of the bitter principles of calamus, acorone and isoacorone, is obtained. PMID- 17405098 TI - Alkaloids from Tabernaemontana psorocarpa. AB - The isolation of the alkaloids from two different samples of stem bark of Tabernaemontana psorocarpa is described. Both samples contained 16-epi isositsirikine as the major alkaloid but differed from each other in the number and quantity of the other alkaloids. The following minor alkaloids were identified: 12-methoxy-14,15-dehydro-vincamine, vallesiachotamine, isovallesiachotamine, tetrahydroalstonine, coronaridine and voacangine. PMID- 17405099 TI - Cycleanine N-oxide, a new alkaloid from Synclisia scabrida. AB - A phytochemical investigation of the stems of Synclisia scabrida Miers (Menispermaceae) resulted in the isolation of a number of known bisbenzylisoquinoline alkaloids: cycleanine, norcycleanine, cocsuline and cocsoline. A new alkaloid, cycleanine N-oxide was also isolated. PMID- 17405100 TI - [Iridoid Glucosides from Euphrasia rostkoviana]. AB - Eight iridoid glucosides have been isolated from Euphrasia rostkoviana by the combination of open column- and high performance liquid chromatography. The structure of a new compound, named eurostoside, and the identity of the others have been established by chemical transformations and spectral evidence of the compounds and their derivatives. PMID- 17405101 TI - Constituents of the chinese crude drug "Sang-Bai-Pi" (morus root bark). AB - From the benzene extract of the Chinese crude drug "Sang-Bai-Pi" (Japanese name Sohakuhi), the root barks of Morus sp. (Moraceae), a novel isoprene substituted flavanone derivative, named Sanggenon B, was isolated; its structure was shown to be I on the basis of spectral and chemical data. Sanggenon B (I) is regarded biogenetically as a variation of a Diels-Alder adduct of a chalcone derivative and a dehydroprenylflavanone derivative. PMID- 17405102 TI - Rapid and simple multiplication of Pinellia ternata by tissue culture. AB - Rapid and simple regeneration of Pinellia ternata has been achieved by tissue culture. The addition of 2,4-dichlorophenoxy acetic acid (2,4-D; 0.5 ppm) and kinetin (1 ppm) stimulated production of a maximum number of regenerated plantlets, while callus formation was induced by concentrations of 2,4-D higher than 1 ppm. alpha-Naphthaleneacetic acid (NAA) was also favourable for regeneration but was less effective than 2,4-D. Growth with indole-3-acetic acid (IAA) and kinetin favoured callus formation in general, but the rate of callus growth was extremely slow. These regenerated plantlets flowered true to their parental strains when transferred to soil and cultivated for 4.5 months. PMID- 17405103 TI - 5-epideoxyloganic acid from Nepeta cataria. AB - On the basis of exhaustive (1)H- and (13)C-NMR spectral studies and chemical transformations, the structure of nepetolglucosylester isolated from Nepeta cataria L. was revised as (5R, 8S, 9R)-7-deoxyloganic acid, which was renamed 5 epideoxyloganic acid. PMID- 17405104 TI - Steroidal saponins from Asparagus curillus fruits. AB - Methanol extract of defatted fruits of Asparagus curillus Buch-Ham yielded a mixture of Saponins, Oligofurostanosides and Spirostanolglycosides. PMID- 17405105 TI - Naphthaquinones of Diospyros batocana. AB - Root bark of West African tree Diospyros batocana Hiern, Ebenaceae, was examined. On the basis of chromatographic and spectral analysis it was shown to contain the naphtaquinones 7-methyljuglone, 2-methylnaphthazarin, biramentaceone, rotundiquinone, mamegakinone, diospyrin, diosquinone, isodiospyrin and batocanone. This last compound proved to be the (-)-enantiomer of isodiospyrin 2', 3'-epoxide. The probable existence of beta-dihydrodiospyrin, of a batocanone isomer and of an isodiospyrin diepoxide in this material is mentioned. The partial synthesis of two isomeric isodiospyrin di-epoxides is described. PMID- 17405106 TI - [Research on alkaloids and isolation of tabersonine from tissue culture of Voacanga thouarsii.]. PMID- 17405107 TI - Isolation of serpentine and tetraphylline from Rauwolfia vomitoria. PMID- 17405108 TI - Pyrazolo[1,5-a]pyrimidines. A combined multinuclear magnetic resonance (1H, 13C, 15N, 19F) and DFT approach to their structural assignment. AB - Multinuclear magnetic resonance spectroscopy together with GIAO-DFT calculations allowed establishment of the structure of the products obtained by condensation of 3(5)-amino-4-phenyl-1H-pyrazole and beta-dicarbonyl compounds bearing a trifluoromethyl group. They are 3-phenyl-5-(R)-7-trifluoromethylpyrazolo[1,5 a]pyrimidines. PMID- 17405109 TI - Computational chemistry development of a unified free energy Markov model for the distribution of 1300 chemicals to 38 different environmental or biological systems. AB - Predicting tissue and environmental distribution of chemicals is of major importance for environmental and life sciences. Most of the molecular descriptors used in computational prediction of chemicals partition behavior consider molecular structure but ignore the nature of the partition system. Consequently, computational models derived up-to-date are restricted to the specific system under study. Here, a free energy-based descriptor (DeltaG(k)) is introduced, which circumvent this problem. Based on DeltaG(k), we developed for the first time a single linear classification model to predict the partition behavior of a broad number of structurally diverse drugs and other chemicals (1300) for 38 different partition systems of biological and environmental significance. The model presented training/predicting set accuracies of 91.79/88.92%. Parametrical assumptions were checked. Desirability analysis was used to explore the levels of the predictors that produce the most desirable partition properties. Finally, inversion of the partition direction for each one of the 38 partition systems evidences that our models correctly classified 89.08% of compounds with an uncertainty of only +/-0.17% independently of the direction of the partition process used to seek the model. Other 10 different classification models (linear, neural networks, and genetic algorithms) were also tested for the same purposes. None of these computational models favorably compare with respect to the linear model indicating that our approach capture the main aspects that govern chemicals partition in different systems. PMID- 17405110 TI - Tetralogy of Fallot in the fetus in the current era. AB - OBJECTIVES: To compare the spectrum of tetralogy of Fallot detected recently in fetal life with that in previous reports of prenatal and postnatal experience. METHODS: All cases of tetralogy of Fallot, including those with pulmonary atresia, diagnosed between 1998 and 2005 inclusive were identified. Additional data for the 129 cases were collected, including associated congenital heart malformations, nuchal translucency measurement, karyotype and outcome. RESULTS: The most common reason for referral was a suspicion of heart malformation at the routine obstetric ultrasonography scan and referrals increased during the study period. The mean gestational age at diagnosis was 20.6 weeks. The nuchal translucency measurement was above the 95th centile in 37 (47%) of the 78 fetuses in which it had been measured, and in 19/37 of the chromosomally normal fetuses. Of 112 fetuses with chromosomal analysis, 55 (49%) had anomalies, including 22q11 microdeletion in 15. There were additional extracardiac malformations in 65/129 cases (50%) and additional cardiac malformations in 73 (57%). In 70/129 (54%) cases, the parents chose termination of pregnancy. Overall survival to date in the continuing pregnancies is 77%. CONCLUSIONS: Tetralogy of Fallot is increasingly recognized during routine fetal obstetric scanning. However, the spectrum of disease detected in the fetus remains biased towards those cases with extracardiac malformations and those with complex disease. As a result, even in the current era, the prognosis for the whole group of fetal cases under the diagnostic heading of tetralogy of Fallot continues to be much less favorable than would be expected of a postnatal series. PMID- 17405111 TI - Prenatal diagnosis and postnatal management of congenital laryngeal atresia in a preterm infant. AB - Laryngeal atresia is a rare congenital cause of high airway obstruction that can lead to death if not correctly recognized and treated at birth. Postnatal management is difficult and the prognosis is often poor. We report a case of prenatal diagnosis of laryngeal atresia in a fetus that was delivered preterm at 29 weeks of gestation. Tracheotomy was performed as an ex utero intrapartum treatment (EXIT) to guarantee patent airway, and laryngotracheoplasty was performed at 22 months of corrected age. A favorable ventilatory and neurodevelopmental outcome was observed at 33 months of age. PMID- 17405112 TI - Growth format, electronic architecture, magnetic, and optical properties of aromatic cyclo-Cu3Au3 homotops. AB - Bimetallic Cu(3)Au(3) clusters have been investigated using electronic structure calculation techniques (DFT) to understand their electronic, magnetic, and optical properties as well as the geometrical structures. The most stable homotop is the planar cyclo-[Cu(3)(micro-Au)(3)] form consisting of a triangular positively charged Cu(3) structural core with negatively charged Au atoms occupying exposed positions. This structure is characterized by the maximum number of heterobonds and peripheral positions of Au atoms. Possible growth formats of the cyclo-[Cu(3)(micro-Au)(3)] homotops have been explored following both the edge-capping and the stepwise metal atom substitution mechanism. The bonding pattern along with the density of states (DOS) plots of the cyclo [Cu(3)(micro-Au)(3)] homotop are thoroughly analyzed and compared with those of the pure cyclo-[Cu(3)(micro-Cu)(3)] and cyclo-[Au(3)(micro-Au)(3)] clusters. Particular attention was paid on the stability of these bimetallic clusters in relation with the ring-shaped electron density distribution (aromaticity). It was found that all 3-membered metal rings exhibit significant aromatic character, which was verified by a number of established criteria of aromaticity, such as structural, energetic, magnetic (NICS profiles), and out-of-plane ring deformability criteria. The NICS (1) values correlate well with the out-of-plane ring deformation energy. Finally, a comprehensive analysis of the optical spectra of the CuAu, Cu(2), and Au(2) diatomics and the cyclo-[Cu(3)(micro-Au)(3)], cyclo [Cu(3)(micro-Cu)(3)], and cyclo-[Au(3)(micro-Au)(3)] clusters placed the electronic assignments of the optical transitions on a firm footing. PMID- 17405113 TI - Explicit treatment of force contribution from alignment tensor using overdetermined linear equations and its application in NMR structure determination. AB - Residual dipolar coupling (RDC) provides valuable information about the orientation of each internuclear vector in a macromolecule with respect to the static magnetic field. However, structure determination utilizing RDC still remains challenging without additional restraints such as NOE. In this context, a novel approach has been developed to efficiently extract structural information from RDC by successive application of singular value decomposition (SVD) method in the course of NMR structure determination. Force contribution from the alignment tensor is rigorously formulated in the context of SVD, and assessments have been made to verify its numerical accuracy. The efficacy of this approach is illustrated by showing that RDC restraints alone can restore a distorted beta hairpin to native-like structure using the replica-exchange molecular dynamics simulations. PMID- 17405114 TI - Calculating rate constants with updated Hessians using variational transition state theory with multidimensional tunneling. AB - Variational transition state theory with multidimensional tunneling (VTST/MT) has been used for calculating the rate constants of reactions. The updated Hessians have been used to reduce the computational costs for both geometry optimization and trajectory following procedures. In this paper, updated Hessians are used to reduce the computational costs while calculating the rate constants applying VTST/MT. Although we found that directly applying the updated Hessians will not generate good vibrational frequencies along the minimum energy path (MEP), however, we can either re-compute the full Hessian matrices at fixed intervals or calculate the Block Hessians, which is constructed by numerical one-side difference for the Hessian elements in the "critical" region and Bofill updating scheme for the rest of the Hessian elements. Due to the numerical instability of the Bofill update method near the saddle point region, we have suggested a simple strategy in which we follow the MEP until certain percentage of the classical barrier height from the barrier top with full Hessians computed and then performing rate constant calculation with the extended MEP using Block Hessians. This strategy results a mean unsigned percentage deviation (MUPD) around 10% with full Hessians computed till the point with 80% classical barrier height for four studied reactions. This proposed strategy is attractive not only it can be implemented as an automatic procedure but also speeds up the VTST/MT calculation via embarrassingly parallelization to a personal computer cluster. PMID- 17405115 TI - Comparison of solvation-effect methods for the simulation of peptide interactions with a hydrophobic surface. AB - In this study we investigated the interaction behavior between thirteen different small peptides and a hydrophobic surface using three progressively more complex methods of representing solvation effects: a united-atom implicit solvation method [CHARMM 19 force field (C19) with Analytical Continuum Electrostatics (ACE)], an all-atom implicit solvation method (C22 with GBMV), and an all-atom explicit solvation method (C22 with TIP3P). The adsorption behavior of each peptide was characterized by the calculation of the potential of mean force as a function of peptide-surface separation distance. The results from the C22/TIP3P model suggest that hydrophobic peptides exhibit relatively strong adsorption behavior, polar and positively-charged peptides exhibit negligible to relatively weak favorable interactions with the surface, and negatively-charged peptides strongly resist adsorption. Compared to the TIP3P model, the ACE and GBMV implicit solvent models predict much stronger attractions for the hydrophobic peptides as well as stronger repulsions for the negatively-charged peptides on the CH(3)-SAM surface. These comparisons provide a basis from which each of these implicit solvation methods may be reparameterized to provide closer agreement with explicitly represented solvation in simulations of peptide and protein adsorption to functionalized surfaces. PMID- 17405116 TI - Quantum correction to the pair distribution function. AB - We report a numerical technique that allows the quantum effects of zero-point motion to be incorporated into Pair Distribution Functions calculated classically for molecules using Monte Carlo or Molecular Dynamics simulations. We establish the basis for this approximation using a diatomic molecule described by a Morse potential. The correction should significantly improve the agreement between modeled and experimental data, and facilitate conclusions about inter- and intra molecular motion and flexibility. We describe a similar approach to obtain the energy and the specific heat. PMID- 17405117 TI - Equivalent potential of water molecules for electronic structure of glutamic acid. AB - The fundamental importance of the electronic structure of molecules is widely recognized. To get reliable electronic structure of protein in aqueous solution, it is necessary to construct a simple, easy-use equivalent potential of water molecules for protein's electronic structure calculation. Here, the first principles, all-electron, ab initio calculations have been performed to construct the equivalent potential of water molecules for the electronic structure of glutamic acid, which is a hydrophilic amino acid and is negatively charged (Glu( )) in neutral water solution. The main process of calculation consists of three steps. Firstly, the geometric structure of the cluster containing Glu(-) and water molecules is calculated by free cluster calculation. Then, based on the geometric structure, the electronic structure of Glu(-) with the potential of water molecules is calculated using the self-consistent cluster-embedding method. Finally, the electronic structure of Glu(-) with the potential of dipoles is calculated. Our calculations show that the major effect of water molecules on Glu(-)'s electronic structure is lowering the occupied electronic states by about 0.017 Ry, and broadening energy gap by 12%. The effect of water molecules on the electronic structure of Glu(-) can be well simulated by dipoles potential. PMID- 17405118 TI - Human papillomavirus type distribution in invasive cervical cancer and high-grade cervical lesions: a meta-analysis update. AB - Data on human papillomavirus (HPV) type distribution in invasive and pre-invasive cervical cancer is essential to predict the future impact of HPV16/18 vaccines and HPV-based screening tests. A meta-analyses of HPV type distribution in invasive cervical cancer (ICC) and high-grade squamous intraepithelial lesions (HSIL) identified a total of 14,595 and 7,094 cases, respectively. In ICC, HPV16 was the most common, and HPV18 the second most common, type in all continents. Combined HPV16/18 prevalence among ICC cases was slightly higher in Europe, North America and Australia (74-77%) than in Africa, Asia and South/Central America (65 70%). The next most common HPV types were the same in each continent, namely HPV31, 33, 35, 45, 52 and 58, although their relative importance differed somewhat by region. HPV18 was significantly more prevalent in adeno/adenosquamous carcinoma than in squamous cell carcinoma, with the reverse being true for HPV16, 31, 33, 52 and 58. Among HSIL cases, HPV16/18 prevalence was 52%. However, HPV 16, 18 and 45 were significantly under-represented, and other high-risk HPV types significantly over-represented in HSIL compared to ICC, suggesting differences in type-specific risks for progression. Data on HPV-typed ICC and HSIL cases were particularly scarce from large regions of Africa and Central Asia. PMID- 17405119 TI - Health-status of adult survivors of childhood cancer: a large-scale population based study from the British Childhood Cancer Survivor Study. AB - The purpose of this study was to investigate the effect of childhood cancer and its treatment on self-reported health-status in 10,189 adult survivors of childhood cancer in Britain. Age- and sex-adjusted scores on the SF-36 Mental and Physical Component Summary scales (MCS, PCS, respectively) were compared between survivors and UK norms, and between subgroups of survivors, by multiple regression. Survivors had comparable scores to UK-norms on the MCS scale (difference (D) = -0.1, 99% CI: -0.5, 0.3). The difference in scores between survivors and UK-norms on the PCS scale varied by age (p(heterogeneity) < 0.001). Young survivors (16-19 years) scored similarly to UK-norms (D = 0.5, (-1.1, 2.2), whereas the age groups of 25 and older scored statistically and clinically significantly below UK-norms (all p-values < 0.0001), with Ds ranging between 2.3 (-3.5, -1.2) and -3.7 (-5.0, -2.4). Survivors of central nervous system (CNS) and bone tumors scored significantly (p-value at all ages <0.003) below UK-norms on the PCS scale. Specifically, these survivors were substantially more limited in specific daily activities such as, for example, walking a mile (40, 63%, respectively) when compared to UK-norms (16%). In conclusion, childhood cancer survivors rate their mental health broadly similarly to those in the general population. Survivors of CNS and bone tumors report their physical health-status to be importantly below population norms. Although self-reported physical health is at least as good as in the general population among young survivors, this study suggests that perceived physical health declines more rapidly over time than in the general population. PMID- 17405120 TI - Multiple myeloma: a review of the epidemiologic literature. AB - Multiple myeloma, a neoplasm of plasma cells, accounts for approximately approximately 15% of lymphatohematopoietic cancers (LHC) and 2% of all cancers in the US. Incidence rates increase with age, particularly after age 40, and are higher in men, particularly African American men. The etiology is unknown with no established lifestyle, occupational or environmental risk factors. Although several factors have been implicated as potentially etiologic, findings are inconsistent. We reviewed epidemiologic studies that evaluated lifestyle, dietary, occupational and environmental factors; immune function, family history and genetic factors; and the hypothesized precursor, monoclonal gammopathies of undetermined significance (MGUS). Because multiple myeloma is an uncommon disease, etiologic assessments can be difficult because of small numbers of cases in occupational cohort studies, and few subjects reporting exposure to specific agents in case-control studies. Elevated risks have been reported consistently among persons with a positive family history of LHC. A few studies have reported a relationship between obesity and multiple myeloma, and this may be a promising area of research. Factors underlying higher incidence rates of multiple myeloma in African Americans are not understood. The progression from MGUS to multiple myeloma has been reported in several studies; however, there are no established risk factors for MGUS. To improve our understanding of the causes of multiple myeloma, future research efforts should seek the causes of MGUS. More research is also needed on the genetic factors of multiple myeloma, given the strong familial clustering of the disease. PMID- 17405121 TI - The non-Hodgkin lymphomas: a review of the epidemiologic literature. AB - The non-Hodgkin lymphomas (NHL) are a heterogeneous group of B-cell and T-cell neoplasms that arise primarily in the lymph nodes. NHL incidence rates in the US doubled between about 1970 and 1990, and stabilized during the 1990s. NHL accounts for approximately 3.4% of cancer deaths in the US. Although some of the observed patterns in NHL have been related to HIV/AIDS, these conditions cannot fully explain the magnitude of the changes; neither do changes in classification systems nor improved diagnostic capabilities. Studies of occupational and environmental exposures (e.g., pesticides, solvents) have produced no consistent pattern of significant positive associations. Inverse associations with ultraviolet radiation exposure and alcohol and fish intake, and positive associations with meat and saturated fat intake have been reported in several studies; additional studies are needed to confirm or refute these associations. Family history of NHL or other hematolympho-proliferative cancers and personal history of several autoimmune disorders are associated with increased risk of NHL, but are not likely to account for a large proportion of cases. HIV and other infectious agents, such as human herpesvirus 8 and Epstein-Barr, appear to be associated with differing types of NHL, such as some B-cell lymphomas. Future epidemiologic studies should evaluate associations by NHL type, enhance exposure information collected, and elucidate factors that may identify susceptible (or resistant) subpopulations because of genetic, immunologic or other characteristics. The extent to which the etiology of NHL types may differ is important to resolve in ongoing and future studies. PMID- 17405122 TI - Serial transplantation of NMU-induced rat mammary tumors: a model of human breast cancer progression. AB - Human breast cancer is a heterogeneous disease that appears to progress from an in situ tumor to invasive cancer. Little is known about the molecular events driving this progression. Although microarray technology has helped us understand the genetic heterogeneity of breast cancer, its application to studying the transition from in situ to invasive disease is limited by the inability to follow the progression of a single patient's tumor. We previously used rat specific microarrays to show that N-methyl-N-nitrosourea induced tumors are similar to low grade estrogen-receptor positive human breast cancer. Here, we transplanted these tumors through 5 generations of syngeneic hosts, and studied 65 resulting tumors. Most transplanted tumors gradually progressed from a noninvasive, low-grade cancer to a higher-grade invasive disease, losing p63 localization and basement membrane integrity. Invasive cancers frequently demonstrated a more mesenchymal phenotype with increased vimentin expression. Additionally, a unique transplant series is described with a phenotype similar to human basal-like breast cancer. Rat-specific Affymetrix gene arrays containing 15,866 gene probes identified genes that differentiated highly invasive tumors from those of low invasive potential. A linear regression analysis was used to find genes whose change in expression paralleled increasing invasive features independent of the transplant lineage of origin. Genes identified were assigned membership in cell adhesion, signal transduction, cell cycle and extracellular matrix groups, among others. This animal model overcomes the difficulty in studying human breast cancer progression. Our data support a gradual and continuous alteration in programs of gene expression during breast cancer invasion. PMID- 17405123 TI - Inductive effects in radicals calculated from DFT energies; substituted bicyclo[2.2.2]octan-1-yloxy radicals. AB - Energies of a series of 4-substituted 1-oxybicyclo[2.2.2]octan-1-yloxy radicals with 18 various substituents were calculated within the framework of the DFT theory at the levels UB3LYP/6-311+G(d,p)//UB3LYP/6-311+G(d,p) and UB3LYP/6 311++G(2df,p)//UB3LYP/6-311+G(d,p) and compared with similar series of the parent alcohols, their deprotonated and protonated forms calculated at the levels B3LYP/6-311+G(d,p)//B3LYP/6-311+G(d,p) and B3LYP/6-311++G(2df,p)//B3LYP/6 311+G(d,p). The two levels are of the same performance and both are sufficient for molecules of this type according to comparison with scarce experimental gas phase acidities and basicities. The substituent effects were analyzed in terms of isodesmic equations. In addition to strong dependence on the substituent inductive effect, a slight dependence on the electronegativity of the first atom of the substituent was proven in certain cases. In all aspects, there is no qualitative difference between the effects on radicals and on similar closed shell species. Radicals behave as slightly electron deficient; the substituent effect is weaker than that on the ions but stronger than on neutral molecules. PMID- 17405124 TI - Speeding up parallel GROMACS on high-latency networks. AB - We investigate the parallel scaling of the GROMACS molecular dynamics code on Ethernet Beowulf clusters and what prerequisites are necessary for decent scaling even on such clusters with only limited bandwidth and high latency. GROMACS 3.3 scales well on supercomputers like the IBM p690 (Regatta) and on Linux clusters with a special interconnect like Myrinet or Infiniband. Because of the high single-node performance of GROMACS, however, on the widely used Ethernet switched clusters, the scaling typically breaks down when more than two computer nodes are involved, limiting the absolute speedup that can be gained to about 3 relative to a single-CPU run. With the LAM MPI implementation, the main scaling bottleneck is here identified to be the all-to-all communication which is required every time step. During such an all-to-all communication step, a huge amount of messages floods the network, and as a result many TCP packets are lost. We show that Ethernet flow control prevents network congestion and leads to substantial scaling improvements. For 16 CPUs, e.g., a speedup of 11 has been achieved. However, for more nodes this mechanism also fails. Having optimized an all-to-all routine, which sends the data in an ordered fashion, we show that it is possible to completely prevent packet loss for any number of multi-CPU nodes. Thus, the GROMACS scaling dramatically improves, even for switches that lack flow control. In addition, for the common HP ProCurve 2848 switch we find that for optimum all to-all performance it is essential how the nodes are connected to the switch's ports. This is also demonstrated for the example of the Car-Parinello MD code. PMID- 17405125 TI - Technical note: revised fully stature estimation technique. PMID- 17405126 TI - Nutritional status of Ribeirinhos in Brazil and the nutrition transition. AB - Anthropometric and household data (size, composition, economic activity) were collected from a population of Ribeirinhos living in a rural setting in the eastern Amazon. Data are compared to international reference standards and to other Amazonian populations with the goals of increasing our understanding of the Amazon's largest ethnic group and identifying the relationship between changes in subsistence strategies and nutritional status. Data on height, weight, skinfolds, and circumferences were collected from 471 adults and subadults. The population showed a high degree of stunting with an average HAZ below -2.0 for all age groups over 3 years, and 60% of adult men and 70% of adult women were stunted. Wasting was rare. Average skinfold thicknesses and upper-arm muscle area were near or below average but within the normal range compared to the reference standard, indicating adequate energy and protein stores. Thirty-one percent of males and 29% of females were overweight/obese, and the highest average BMIs were found among men and women in their 40s. Adult males who participated in wage labor had higher weights, BMIs, and UMA values, and were more likely to be overweight and obese compared with those who did not work in wage-labor jobs. Children of fathers who worked in wage labor had higher BMI and UMA values, but there was no significant effect on the nutritional status of other adults in these same households. Signs of the nutrition transition were most noticeable among adult males involved in wage labor because of changes in their diet and activity patterns. PMID- 17405129 TI - N-(4-(4-(2-Halogenophenyl)piperazin-1-yl)butyl) substituted cinnamoyl amide derivatives as dopamine D2 and D3 receptor ligands. AB - A series of eight substituted N-(4-(4-(2-halogenophenyl)piperazin-1-yl)butyl)-3 phenylacryl amide derivatives have been synthesized and screened for binding affinities at dopamine hD(2) and hD(3) receptors. All compounds have shown high to remarkable receptor affinities and some have led to distinct selectivity for D(3) receptors. Highest D(3)-receptor affinity has been observed for 3-(4 aminophenyl)-N-(4-(4-(2-fluorophenyl)piperazin-1-yl)butyl)acryl amide (hD(3) K(i) 0.9 nM; hD(2) K(i) 17.4 nM). Selectivity ratio has been best for 3-(4 chlorophenyl)-N-(4-(4-(2-fluorophenyl)piperazin-1-yl)butyl)acryl amide with a 56 fold preference for hD(3) versus hD(2). A functional activity test has been performed by a mitogenesis test for N-(4-(4-(2-fluorophenyl)piperazin-1-yl)butyl) 3,3-diphenylacryl amide, which, surprisingly, has shown full agonist properties. PMID- 17405130 TI - Homology modeling of the serotonin transporter: insights into the primary escitalopram-binding site. AB - The serotonin transporter (SERT) is one of the neurotransmitter transporters that plays a critical role in the regulation of endogenous amine concentrations and therefore is an important target for therapeutic agents affecting the central nervous system. The recently published, high resolution X-ray structure of the closely related amino acid transporter, Aquifex aeolicus leucine transporter (LeuT), provides an opportunity to develop a three-dimensional model of the structure of SERT. We present herein a homology model of SERT using LeuT as the template and containing escitalopram as a bound ligand. Our model explains selectivities known from mutational studies and varying ligand data, which are discussed and illustrated in the paper. PMID- 17405131 TI - Night blindness-associated mutations in the ligand-binding, cysteine-rich, and intracellular domains of the metabotropic glutamate receptor 6 abolish protein trafficking. AB - Mutations in the GRM6 gene, which encodes the metabotropic glutamate receptor 6 (mGluR6), lead to autosomal recessive congenital stationary night blindness (CSNB), which is characterized by loss of night vision due to a defect in signal transmission from photoreceptor to the adjacent ON-bipolar cells in the retina. So far, the sequence variations that have been described in six different families include nonsense, frameshift, and missense mutations. Here we investigated the impact of missense mutations in the ligand-binding domain, a conserved cysteine-rich domain, and the intracellular domain on the localization of the protein. We visualized and discriminated between surface and intracellular protein. Here we demonstrate that the wild-type (wt) protein localizes to the cell surface, and to endoplasmic reticulum (ER) and Golgi compartments. This also holds true for a mGluR6 variant containing a polymorphic, nondisease-associated amino acid exchange in the ligand-binding domain. In contrast, all disease associated missense mutations lead to retention of the protein in the ER, while dimerization seems not to be affected. This is the first report that shows that CSNB-associated mutations in three different domains of mGluR6 abolish proper protein trafficking. We propose that the ligand-binding and the poorly characterized cysteine-rich domains, in addition to the intracellular domains, have a pivotal role in correct trafficking of metabotropic glutamate receptors to the cell surface. PMID- 17405132 TI - Molecular and in silico analyses of the full-length isoform of usherin identify new pathogenic alleles in Usher type II patients. AB - The usherin gene (USH2A) has been screened for mutations causing Usher syndrome type II (USH2). Two protein isoforms have been identified: a short isoform of 1,546 amino acids and a more recently recognized isoform extending to 5,202 amino acids. We have screened the full length by genomic sequencing. We confirm that many mutations occur in the exons contributing solely to the longer form. USH2 is an autosomal recessive disorder and, in contrast to previous studies, both mutations were identified in 23 patients and a single mutation in 2 out of 33 patients. A total of 34 distinct mutated alleles were identified, including one complex allele with three variants and another with two. A total of 27 of these are novel, confirming that most mutations in usherin are private. Many of the mutations will lead to prematurely truncated protein but as there are a substantial number of missense variants, we have used in silico analysis to assess their pathogenicity. Evidence that they are disease-causing has been produced by protein alignments and three-dimensional (3D) structural predictions when possible. We have identified a previously unrecognized cysteine rich structural domain, containing 12 dicysteine repeats, and show that three missense mutations result in the loss of one of a pair of the defining cysteine-cysteine pairs. PMID- 17405133 TI - Mass spectrometry for enzyme assays and inhibitor screening: an emerging application in pharmaceutical research. AB - Robust methods that monitor enzyme activity and inhibitor potency are crucial to drug discovery and development. Over the past 20 years, mass spectrometric methods have increasingly been used to measure enzyme activity and kinetics. However, for rapid screening of inhibitory compounds, various forms of fluorescence and chemiluminscence readout have continued to dominate the market. As the sensitivity, speed, and miniaturization of mass spectrometry methods continue to advance, opportunities to couple mass spectrometry with screening will continue to come to the forefront. To appreciate the tremendous potential for MS-based screening assays, it becomes necessary to understand the current state of capabilities in this arena. Thus, this review is intended to capture how mass spectrometry for studying enzymes activity has progressed from simple qualitative questions (i.e., is the product detected?) to quantitative measures of enzyme activity and kinetics and then as a tool for rapidly screening inhibitory compounds as an alternative to current methods of high throughput drug screening. PMID- 17405135 TI - Collagen turnover in the adult femoral mid-shaft: modeled from anthropogenic radiocarbon tracer measurements. AB - We have measured the (14)C content of human femoral mid-shaft collagen to determine the dynamics of adult collagen turnover, using the sudden doubling and subsequent slow relaxation of global atmospheric (14)C content due to nuclear bomb testing in the 1960s and 1970s as a tracer. (14)C measurements were made on bone collagen from 67 individuals of both sexes who died in Australia in 1990 1993, spanning a range of ages at death from 40 to 97, and these measurements were compared with values predicted by an age-dependent turnover model. We found that the dataset could constrain models of collagen turnover, with the following outcomes: 1) Collagen turnover rate of females decreases, on average, from 4%/yr to 3%/yr from 20 to 80 years. Male collagen turnover rates average 1.5-3%/yr over the same period. 2) For both sexes the collagen turnover rate during adolescent growth is much higher (5-15%/yr at age 10-15 years), with males having a significantly higher turnover rate than have females, by up to a factor of 2. 3) Much of the variation in residual bomb (14)C in a person's bone can be attributed to individual variation in turnover rate, but of no more than about 30% of the average values for adults. 4) Human femoral bone collagen isotopically reflects an individual's diet over a much longer period of time than 10 years, including a substantial portion of collagen synthesised during adolescence. PMID- 17405136 TI - Myelinated nerve endings in human skin. AB - We used immunohistochemical techniques and confocal microscopy to study the morphometry of myelinated nerve endings in glabrous and hairy skin. A total of 30 healthy volunteers took part in this study designed to assess the possibility of obtaining reliable information on myelinated fibers using samples of hairy skin and to determine whether differences exist between myelinated terminations from different sites. We obtained consistent information on cutaneous myelinated terminations using hairy as well as glabrous skin samples. Myelinated endings from hairy and glabrous skin differ in density and distribution. However, from a comparison of our findings with data from nerve biopsy studies, we conclude that all cutaneous myelinated terminations are thinner terminal branches of large myelinated A beta fibers, whereas cutaneous terminations of small myelinated A delta fibers lose their myelin before entering the dermis and become indistinguishable from C-fiber terminations. The classic criteria, based on fiber size, used to distinguish myelinated fiber subgroups in sensory nerves are therefore not suitable for identifying myelinated terminations in the skin. PMID- 17405137 TI - Satoyoshi syndrome has antibody against brain and gastrointestinal tissue. AB - Satoyoshi syndrome is a rare postnatal disorder with muscle spasms, alopecia, and diarrhea of unknown etiology. Nutritional deficiency seems to influence lifespan. We present a patient with this syndrome having a unique "mesh-like" mucosal change radiographically and white granules endoscopically in the gastrointestinal tract. A common antibody against brain, stomach, and duodenal tissue, according to Western blot analysis, was detected in the sera of two patients with this syndrome. These findings suggest that Satoyoshi syndrome is a systemic autoimmune disease involving the nervous, endocrine, and gastrointestinal systems. PMID- 17405138 TI - Accuracy of repetitive nerve stimulation for diagnosis of the cramp-fasciculation syndrome. AB - The goal of this retrospective cohort study was to test the hypothesis that the cramp-fasciculation syndrome (CFS) represents a disorder of peripheral nerve hyperexcitability and to evaluate the accuracy of repetitive nerve stimulation (RNS) for its diagnosis. A consecutive series of 108 patients were evaluated with posterior tibial RNS at 1, 2, and 5 HZ. Abnormal peripheral nerve excitability was defined by the presence of afterdischarges, cramp potentials, or continuous motor unit activity. RNS demonstrated abnormal nerve hyperexcitability in 29 of 36 subjects (81%) with CFS, defined operationally by the presence of both muscle cramps and fasciculations. Based on receiver operating characteristic (ROC) curve analysis, tibial RNS correctly classified the presence or absence of CFS in 75% of subjects. These results suggest that CFS represents a form of peripheral nerve hyperexcitability and, furthermore, that RNS is a clinically useful test for CFS. PMID- 17405139 TI - Axial myopathy due to primary amyloidosis. AB - A 74-year-old man presented with progressive weakness involving only his back muscles. Serum protein electrophoresis revealed the presence of a monoclonal gammopathy. Muscle biopsy of affected muscles demonstrated amyloid light chain deposition surrounding individual muscle fibers as well as in the walls of blood vessels in association with vasculitis. Selective involvement of axial muscles in isolated amyloid myopathy has not been previously described. We report the occurrence of axial myopathy with associated vasculitis as a presenting feature of primary (AL) amyloidosis. Amyloidosis should be considered in the differential diagnosis of focal myopathies, since the condition may be responsive to chemotherapy. PMID- 17405140 TI - Persistent multifocal conduction block in vasculitic neuropathy with IgM anti gangliosides. AB - A 30-year-old man with essential cryoglobulinemia presented with an axonal neuropathy and was found to have vasculitis at nerve biopsy. After 44 months, in accord with clinical deterioration, motor conduction studies showed excessive temporal dispersion multifocally, with partial conduction block persisting for 3 years. Antibody testing showed the presence of IgM anti-GM1, anti-GD1a, and anti GM2 antibodies. Transitory conduction block has been reported occasionally in patients with vasculitis. The persistent multifocal conduction abnormalities found in this patient were more likely due to a superimposed immunomediated demyelination rather than to chronic nerve ischemia secondary to vasculitis. PMID- 17405141 TI - A novel missense mutation in the caveolin-3 gene in rippling muscle disease. AB - Rippling muscle disease (RMD) is a benign myopathy with symptoms and signs of muscular hyperirritability. We report a 17-year-old patient who presented with muscular hypertrophy, local mounding on percussion, and a rippling phenomenon. Needle electromyography showed electrical silence during the rippling phenomenon. Muscle protein immunohistochemical analysis showed a partial deficiency of caveolin-3. Molecular analysis revealed a novel heterozygous A>C transition at nucleotide position 140 in exon 2 of the caveolin-3 gene. We associated this novel mutation with RMD. PMID- 17405142 TI - Cervical myelopathy caused by retrograde intraneural dissection of anesthetic solution. AB - Brachial plexus block is a generally safe procedure for postoperative analgesia following shoulder surgery. This report describes brachial plexus block complicated by intraneural dissection of anesthetic solution, resulting in severe damage to the upper spinal cord. Other investigators have hypothesized that this rare complication is caused by accidental needle placement within the substance of the cord. This case demonstrates that the complication can occur despite a peripherally located needle. PMID- 17405143 TI - Mass spectrometry-based "omics" technologies in cancer diagnostics. AB - Many "omics" techniques have been developed for one goal: biomarker discovery and early diagnosis of human cancers. A comprehensive review of mass spectrometry based "omics" approaches performed on various biological samples for molecular diagnosis of human cancers is presented in this article. Furthermore, the existing and potential problems/solutions (both de facto experimental and bioinformatic challenges), and future prospects have been extensively discussed. Although the use of present omic methods as diagnostic tools are still in their infant stage and consequently not ready for immediate clinical use, it can be envisaged that the "omics"-based cancer diagnostics will gradually enter into the clinic in next 10 years as an important supplement to current clinical diagnostics. PMID- 17405144 TI - Analytical strategies for identifying drug metabolites. AB - With the dramatic increase in the number of new chemical entities (NCEs) arising from combinatorial chemistry and modern high-throughput bioassays, novel bioanalytical techniques are required for the rapid determination of the metabolic stability and metabolites of these NCEs. Knowledge of the metabolic site(s) of the NCEs in early drug discovery is essential for selecting compounds with favorable pharmacokinetic credentials and aiding medicinal chemists in modifying metabolic "soft spots". In development, elucidation of biotransformation pathways of a drug candidate by identifying its circulatory and excretory metabolites is vitally important to understand its physiological effects. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) have played an invaluable role in the structural characterization and quantification of drug metabolites. Indeed, liquid chromatography (LC) coupled with atmospheric pressure ionization (API) MS has now become the most powerful tool for the rapid detection, structure elucidation, and quantification of drug-derived material within various biological fluids. Often, however, MS alone is insufficient to identify the exact position of oxidation, to differentiate isomers, or to provide the precise structure of unusual and/or unstable metabolites. In addition, an excess of endogenous material in biological samples often suppress the ionization of drug-related material complicating metabolite identification by MS. In these cases, multiple analytical and wet chemistry techniques, such as LC-NMR, enzymatic hydrolysis, chemical derivatization, and hydrogen/deuterium-exchange (H/D-exchange) combined with MS are used to characterize the novel and isomeric metabolites of drug candidates. This review describes sample preparation and introduction strategies to minimize ion suppression by biological matrices for metabolite identification studies, the application of various LC-tandem MS (LC MS/MS) techniques for the rapid quantification and identification of drug metabolites, and future trends in this field. PMID- 17405145 TI - Reduced raft-association of NF155 in active MS-lesions is accompanied by the disruption of the paranodal junction. AB - Neurofascin155 (NF155) is required for the establishment of the paranodal axo glial junction, the predominant interaction site between myelin and axon. It has been shown that the distribution of NF155 is altered in demyelinating diseases such as multiple sclerosis (MS). However, little is known about the biochemical mechanisms underlying these changes. We therefore compared NF155 in postmortem tissue of active and chronic inactive MS lesions with white matter from healthy controls. Although NF155 showed a very similar expression in all control white matter samples, a strong individual variation was observed in MS-lesions with NF155-levels reduced in most samples. At the same time an NF155-fragment was increased in MS-lesions, suggesting that NF155 is subject to protein degradation in lesion sites. Interestingly, the association of NF155 to membrane microdomains (rafts) was reduced in all lesions, irrespective of the amount of NF155, indicating that membrane association of NF155 was generally affected. Therefore, myelin fractionation experiments were performed to analyze the fate of paranodal proteins during demyelination. Although NF155 was enriched in heavy myelin from both control white matter and active MS-lesions, association of Caspr1/paranodin with heavy myelin was abolished in MS-lesions, demonstrating that paranodal junctions are disrupted. In conclusion, the data support the hypothesis that efficient raft-association of NF155 is essential for the assembly of the paranodal junction and demonstrate that reduced association of NF155 to lipid rafts is accompanied by the disassembly of the paranodal junction and thus contributes to the demyelination process in MS. PMID- 17405146 TI - N-WASP regulates extension of filopodia and processes by oligodendrocyte progenitors, oligodendrocytes, and Schwann cells-implications for axon ensheathment at myelination. AB - The molecular mechanisms used by oligodendrocyte precursor cells (OPCs), oligodendrocytes (OLs), and Schwann cells (SCs) to advance processes for motility in the developing nervous system and to ensheath axons at myelination are currently not well defined. Here we demonstrate that OPCs, OLs, and SCs express the major proteins involved in actin polymerization-driven protrusion; these key proteins including F-actin, the Arp2/3 complex, neural-Wiskott Aldrich Syndrome protein (N-WASP) and WAVE proteins, and the RhoGTPases Rac and Cdc42 are present at the leading edges of processes being extended by OPCs, OLs, and SCs. We reveal by real-time PCR that OLs and SCs have different dominant WAVE isoforms. Inhibition of the WASP/WAVE protein, N-WASP, with wiskostatin that prevents activation of the Arp2/3 complex, blocks process extension by OPCs and SCs. Inhibition of N-WASP also causes OPC and SC process retraction, which is preceded by retraction of filopodia. This implicates filopodia in OPC and SC process stability and also of N-WASP in OPC and SC process dynamics. We also demonstrate that p34 (a component of the Arp2/3 complex), WASP/WAVE proteins, actin, alpha tubulin, Rac, Cdc42, vinculin, and focal adhesion kinase are detected in water shocked myelin purified from brain. Inhibition of N-WASP with wiskostatin decreases the number of axons undergoing initial ensheathment in intact optic nerve samples and reduces the Po content of dorsal root ganglia:SC co-cultures. Our findings indicate that OPCs, OLs, and SCs extend processes using actin polymerization-driven protrusion dependent on N-WASP. We hypothesize that inner mesaxons of OLs and SCs use the same mechanism to ensheath axons at myelination. PMID- 17405147 TI - Olfactory ensheathing cells promote migration of Schwann cells by secreted nerve growth factor. AB - Transplantation of Schwann cells (SCs) and olfactory ensheathing cells (OECs) have emerged as very promising therapies for spinal cord repair. The important features of interaction between SCs and OECs are beginning to be appreciated, although the underlying mechanism remains unclear. In the present study, we tested the effects of OECs on SCs migration using a range of in vitro migration assays. We found that SCs migrated abundantly upon OECs monolayer, and the migration-promoting effects were identified to be due to the secreted diffusible factors in OEC-derived conditioned medium (OEC-CM). Furthermore, neutralizing nerve growth factor (NGF) in OEC-CM with NGF antibody could block this effect. Moreover, we found that NGF promotes SCs migration even on astrocyte monolayer. Taken together, these findings provide the first evidence that OECs can promote SCs migration in astrocytic environment by secreted NGF. PMID- 17405148 TI - Purines induce directed migration and rapid homing of microglia to injured pyramidal neurons in developing hippocampus. AB - Traumatic CNS injury activates and mobilizes resident parenchymal microglia (MG), which rapidly accumulate near injured neurons where they transform into phagocytes. The mechanisms underlying this rapid 'homing' in situ are unknown. Using time-lapse confocal imaging in acutely excised neonatal hippocampal slices, we show that rapid accumulation of MG near somata of injured pyramidal neurons in the stratum pyramidale (SP) results from directed migration from tissue regions immediately adjacent to (<200 microm from) the SP. Time-lapse sequences also reveal a 'spreading activation wave' wherein MG situated progressively farther from the SP begin to migrate later and exhibit less directional migration toward the SP. Because purines have been implicated in MG activation and chemotaxis, we tested whether ATP/ADP released from injured pyramidal neurons might account for these patterns of MG behavior. Indeed, application of apyrase, which degrades extracellular ATP/ADP, inhibits MG motility and homing to injured neurons in the SP. Moreover, bath application of exogenous ATP/ADP disrupts MG homing by inducing directional migration toward the slice exterior and away from injured neurons. These results indicate that extracellular ATP/ADP is both necessary and sufficient to induce directional migration and rapid homing of neonatal MG to injured neurons in situ. Rapid, ATP/ADP-dependent MG homing may promote clearance of dead and dying cells and help limit secondary damage during the critical first few hours after neuronal injury. PMID- 17405149 TI - Expression of a functional metabotropic glutamate receptor 5 on enteric glia is altered in states of inflammation. AB - The metabotropic glutamate receptor 5 (mGluR5) is expressed by astrocytes and its expression is modulated by inflammation. Enteric glia have many similarities to astrocytes and are the most numerous cell in the enteric nervous system (ENS). We investigated whether enteric glia express a functional mGluR5 and whether expression of this receptor was altered in colitis. In both enteric plexuses of the ileum and colon of guinea pigs and mice, we observed widespread glial mGluR5 expression. Incubation of isolated segments of the guinea pig ileum with the mGluR5 specific agonist RS-2-chloro-5-hydroxyphenylglycine (CHPG) caused a dose dependent increase in the glial expression of c-Fos and the phosphorylated form of the extracellular signal-regulated kinase 1/2. Preincubation of tissues with the group I metabotropic glutamate receptor antagonist, S-4-carboxyphenylglycine, abolished the effects of CHPG. We examined mGluR5 expression in the guinea pig trinitrobenzene sulfonic acid and the IL-10 gene-deficient (IL-10(-/-)) mouse models of colitis. In guinea pigs, mGluR5 immunoreactivity became diffusely localized over the colonic myenteric ganglia, suggesting a change in receptor distribution. In contrast, glial mGluR5 expression was significantly reduced in the colonic myenteric plexus of IL-10(-/-) mice, as assessed with both real-time quantitative RT-PCR as well as immunohistochemistry and image analysis. These changes occurred without concomitant changes to enteric ganglia or glial fibrillary acidic protein expression in the IL-10(-/-) mouse. Our data suggest that enteric glia are a functional target of the glutamatergic neurotransmitter system in the ENS and that changes in mGluR5 expression may be of physiological significance during colitis. PMID- 17405150 TI - Identification and characterization of a novel RPGR isoform in human retina. AB - Retinitis pigmentosa (RP) constitutes a major cause of blindness and the Retinitis Pigmentosa GTPase Regulator (RPGR) gene accounts for up to 80% of all X linked RP cases. A novel isoform of RPGR, expressed in the human retina, was identified and characterized. It truncates the Regulator of Chromosome Condensation 1 (RCC1) homologous protein domain (RCC1h) of RPGR and mediates the formation of isoform-specific complexes with the RPGR-interacting protein 1 (RPGRIP1). Immunohistochemistry localized the novel RPGR isoform predominantly to inner segments of cone photoreceptors, where it colocalizes with RPGRIP1 in the human retina. In a patient with a mild RP phenotype, we identified a nucleotide substitution in a splicing regulator, which leads to 3.5 times higher levels of the transcripts coding for the novel RPGR isoform. The nucleotide substitution affects regulated alternative splicing of the novel RPGR isoform and suggests a tight adjustment of splicing as a prerequisite for proper function of photoreceptors. PMID- 17405151 TI - Late cardiac effects of anthracycline containing therapy for childhood acute lymphoblastic leukemia. AB - BACKGROUND: At present about 80% of children with acute lymphoblastic leukemia (ALL) will be cured following treatment with multi-drug chemotherapy. A major concern for this growing number of survivors is the risk of late effects of treatment. The aim of this study was to determine whether signs of cardiomyopathy were present in patients treated in childhood with cumulative anthracycline doses of less than 300 mg/m(2). PROCEDURE: Evaluation of cardiac function in a cohort of 63 long-term survivors in first continuous remission following treatment of ALL with multi-drug chemotherapy including anthracyclines was performed using standard M-mode echocardiography and tissue doppler imaging (TDI). Associations between age at diagnosis, cumulative dose of anthracycline, sex, length of follow up, and deviations from normal values in M-mode echocardiograms were evaluated using univariate and multivariate regression analysis. TDI data were compared to normal values using Wilcoxon matched-pairs signed-ranks test. RESULTS: By standard M-mode echocardiography the most significant findings were diastolic dilation of the left ventricle, thinner interventricular septum (IVS), decreased left ventricular mass (LVM) in females, follow-up dependent dilation of the left ventricle in systole and follow-up dependent decrease in ejection fraction (EF). TDI abnormalities included signs of early diastolic dysfunction and myocardial hypertrophy, and were also found in structures that appeared normal by M-mode echocardiography. CONCLUSIONS: This study adds to the growing evidence that even low to moderate doses of anthracyclines might lead to progressive cardiac dysfunction. It is important that children treated with anthracyclines receive life long follow-up for signs of cardiomyopathy. PMID- 17405152 TI - The effect of Ginkgo biloba on the cerebellum of aging SAMP mouse--a TUNEL, bcl 2, and fMRI study. AB - EGb 761, an extract from Ginkgo biloba that possesses neuroprotective properties, was fed to a strain of fast aging mice (SAMP-8) beginning at 3 weeks of age until they were sacrificed at 3 months and 11 months, respectively, along with an age matched control group without herbal feeding. The aim of the study was to determine (1) the status of apoptosis and the status of bcl-2, a molecule involved in the fate of cells following injury, in the cerebella of these mice and (2) to analyze the functional changes as shown by fMRI images. The data indicated that there were no differences in apoptosis between the mice fed with EGb 761 and the control group at the two time points of 3 and 11 months of age. For bcl-2 positive cells, there was a decrease in density only in the cerebella of 11-month-old mice fed with the herbal extract when compared with controls. Functional studies indicated that while no changes were observed in the 3-month old mice fed with Ginkgo biloba, an expansion of activated sites, possibly related to "synaptic reorganization and pathway alteration," was observed in the 11-month-old mice. PMID- 17405153 TI - Proteomics in neurosciences. AB - This review provides an outline of the most important proteomic applications in the study of neurodegenerative disorders including Alzheimer's (AD), Parkinson's (PD), Huntington's (HD), and prion diseases, and also discusses advances in cancer and addiction. One of the scopes is to illustrate the potential of proteomics in the biomarkers discovery of these diseases. Finally, this article comments the advantages and drawbacks of the most commonly used techniques and methods for samples preparation. PMID- 17405154 TI - The clinical significance of tumor grade in non-rhabdomyosarcoma soft tissue sarcomas. PMID- 17405155 TI - Colorectal carcinoma in children and adolescents: the experience of the Istituto Nazionale Tumori of Milan, Italy. AB - BACKGROUND: Colorectal carcinoma (CRC) is one of the most common tumors in adults, but extremely rare in young age. This study retrospectively reports on a group of 27 patients <30 years of age, and particularly on 7 cases <18 years old, treated at the Istituto Nazionale Tumori, Milan, Italy, between 1985 and 2005. PATIENTS AND METHODS: Among the children/adolescents (age 9-18, median 12 years), 5/7 had unfavorable CRC histotypes (poorly differentiated or mucinous adenocarcinoma) and all but one had advanced disease at onset. Initial surgical resection was complete in 5/7 cases, and all patients received postoperative chemotherapy. RESULTS: In the subset of patients <18 years, 6/7 had tumor progression or relapse, and 5 died of their tumor: overall survival (OS) was 23% at 5 years. In the group of 19- to 29-year-olds (young adults), 5-year OS was 72.6%. CONCLUSIONS: This study confirms the rarity and poor prognosis of CRC in children and adolescents: advanced stage and an aggressive biology are hallmarks of this tumor in pediatric age, while clinical findings and outcome in young adults seem more similar to those observed in adult series. Therapeutic recommendations should stay the same as for adults. Surgery remains the mainstay of treatment and early diagnosis is crucial: it is important for pediatricians to be aware that CRC does occur in children, in order to refer suspected cases to expert physicians professionally dedicated to the management of this cancer in adults. PMID- 17405156 TI - Complex chromosome 8;21 translocation with associated hyperdiploidy in acute myeloid leukemia (FAB-M2). AB - We present a case of acute myeloblastic leukemia (AML-M2) with a complex t(8;21) translocation and additional acquired chromosomes yielding a hyperdiploid karyotype. AML1/ETO transcript was observed by reverse transcription-polymerase chain reaction. Fluorescence in situ hybridization (FISH), spectral karyotyping (SKY), and comparative genomic hybridization (CGH) were performed to further identify the chromosomes observed by G banding. The patient was treated according to our current protocol for AML. He remains in complete remission +11 months from diagnosis. Further follow-up of this patient and the analysis of a larger number of children are needed to define whether the gains of the specific extra chromosomes modify the good prognosis that t(8;21) confers to this subgroup of AML. PMID- 17405157 TI - A case of anaplastic large cell lymphoma-induced hemophagocytic lymphohistiocytosis in an adolescent female. PMID- 17405158 TI - Long-term follow-up of metal-on-metal total hip replacement. AB - Metal-on-metal articulations were reintroduced to reduce polyethylene particle induced osteolysis. Elevated serum metal levels have been detected at short- and intermediate-term follow-up. There is little knowledge about long-term effects of increased ionic metal generation. Our study was undertaken to provide information about long-term survival, clinical outcome, radiographic appearance, and serum metal concentrations after a minimum 10 years follow-up with a metal-on-metal hip prosthesis: 105 cementless primary total hip prostheses with metal-on-metal articulating surfaces were implanted in 98 patients with a mean age of 56 years. Clinical data, radiographs, and blood samples were obtained at a follow-up visit 10 years after implantation. Serum cobalt and chromium levels were determined with use of atomic absorption spectrometry. General laboratory analyses included a full blood count and kidney function parameters. The Harris score was 92 points and the UCLA score 6 points after 10 years. Small osteolytic lesions and radiolucent lines were found in Gruen's zones 1, 7, 8, and 14. The probability of survival was 98.6%. The median serum cobalt concentration of the 22 patients with their hip replacement as the only source of cobalt was 0.75 microg/L (range, 0.3 50.1 microg/L). No patient was diagnosed with renal insufficiency during the study period. Five patients were diagnosed with a malignancy between surgery and the follow-up. We do not have evidence of an increased rate of primary malignancies nor could we detect renal failure in our study group. Serum metal levels did not differ from short- and intermediate term follow-up values. PMID- 17405159 TI - Quantum chemical investigation of hydrogen-bond strengths and partition into donor and acceptor contributions. AB - We present a simple increment model for use in the rapid scoring of hydrogen bond strengths employing 15 chemically diverse donor and 28 acceptor terms. The increments cover a large variety of hydrogen bond donor and acceptor groups and are more specific than SYBYL atom types. The increments have been fitted to quantum chemical ab initio interaction energies of 81 small hydrogen-bonded complexes determined at the level of second-order Moller-Plesset perturbation theory (MP2). The complexes have been chosen such as to represent the most important types of donor-acceptor pairs found in biological systems. Sulphur is found to be a strong hydrogen bond acceptor while its donor capacities are weak. By taking CH acidic H donors into account, a linear correlation between MP2 energies and the increment model with a coefficient of correlation of r2=0.994 has been accomplished. The transferability of the fitted parameters has been assessed on a second set of complexes including larger molecules of biological relevance. Very good agreement has been achieved for noncyclic hydrogen bonds. Cooperative effects are not accounted for by the current increment model. For this reason, binding energies of strong cyclic hydrogen bonds, as e.g. present in DNA base pairs, are underestimated by about 30-40%. PMID- 17405160 TI - Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. AB - Mesenchymal stem cells (MSCs) provide an important source of pluripotent cells for musculoskeletal tissue repair. This study examined the impact of MSC implantation on cartilage healing characteristics in a large animal model. Twelve full-thickness 15-mm cartilage lesions in the femoropatellar articulations of six young mature horses were repaired by injection of a self-polymerizing autogenous fibrin vehicle containing mesenchymal stem cells, or autogenous fibrin alone in control joints. Arthroscopic second look and defect biopsy was obtained at 30 days, and all animals were euthanized 8 months after repair. Cartilage repair tissue and surrounding cartilage were assessed by histology, histochemistry, collagen type I and type II immunohistochemistry, collagen type II in situ hybridization, and matrix biochemical assays. Arthroscopic scores for MSC implanted defects were significantly improved at the 30-day arthroscopic assessment. Biopsy showed MSC-implanted defects contained increased fibrous tissue with several defects containing predominantly type II collagen. Long-term assessment revealed repair tissue filled grafted and control lesions at 8 months, with no significant difference between stem cell-treated and control defects. Collagen type II and proteoglycan content in MSC-implanted and control defects were similar. Mesenchymal stem cell grafts improved the early healing response, but did not significantly enhance the long-term histologic appearance or biochemical composition of full-thickness cartilage lesions. PMID- 17405161 TI - Body vigilance: bi-national examination of the construct. AB - Two inter-related studies evaluated the measurement model and construct validity of body vigilance, as indexed by the body vigilance scale [BVS; Schmidt et al., 1997: J Consulting Clin Psychol 65:214-220]. Confirmatory factor analyses (CFA) and convergent, discriminant, and incremental tests of validity of body vigilance were conducted among separate nonclinical samples from the United States (US) (study 1) and Spain (study 2). In both the US and Spanish samples, poor fit for the four-item unidimensional measurement model of body vigilance was observed; good fit, however, was found for a unidimensional, three-item measurement model. Subsequent analyses demonstrated both theoretically predicted as well as a number of unexpected associations between body vigilance and a variety of theoretically relevant external criterion variables. Results are discussed in terms of their clinical implications for advancing assessment of body vigilance and theoretical implications for better understanding the nomological nature of the construct. PMID- 17405162 TI - Investigation of an attentional bias for fear-related material in obsessive compulsive checkers. AB - Prior studies on attentional biases in obsessive-compulsive disorder (OCD) have yielded conflicting results. Using a new methodology based on a spatial cueing paradigm, it was investigated whether OCD checkers display heightened vigilance for concern-related material or difficulties disengaging from such stimuli. Twenty-eight OCD patients, 21 of whom were checkers, were compared to 27 controls. In the spatial cueing paradigm task, patients and controls responded to a visual target at one out of two possible locations, which was preceded by a verbal cue word presented at the same or at the opposite location of the target. Cue words were either checking-relevant (e.g., doubt), neutral (e.g., box), or paranoia-related (e.g., spy). No evidence for an attentional bias in OCD checkers was found. This study is in accordance with previous studies, which were unable to detect attentional biases in OCD patients. PMID- 17405163 TI - Maternal caffeine consumption and risk of cardiovascular malformations. AB - BACKGROUND: The physiologic effects and common use of caffeine during pregnancy call for examination of maternal caffeine consumption and risk of birth defects. Epidemiologic studies have yielded mixed results, but such studies have grouped etiologically different defects and have not evaluated effect modification. METHODS: The large sample size and precise case classification of the National Birth Defects Prevention Study allowed us to examine caffeine consumption and specific cardiovascular malformation (CVM) case groups. We studied consumption of caffeinated coffee, tea, soda, and chocolate to estimate total caffeine intake and separately examined exposure to each caffeinated beverage. Smoking, alcohol, vasoactive medications, folic acid supplement use, and infant gender were evaluated for effect modification. Maternal interview reports for 4,196 CVM case infants overall and 3,957 control infants were analyzed. RESULTS: We did not identify any significant positive associations between maternal caffeine consumption and CVMs. For tetralogy of Fallot, nonsignificant elevations in risk were observed for moderate (but not high) caffeine intake overall and among nonsmokers (ORs of 1.3 to 1.5). Risk estimates for both smoking and consuming caffeine were less than the sum of the excess risks for each exposure. We observed an inverse trend between coffee intake and risk of atrial septal defect; however, this single significant pattern of association might have been a chance finding. CONCLUSIONS: Our study found no evidence for an appreciable teratogenic effect of caffeine with regard to CVMs. PMID- 17405164 TI - Role of annexin 1 gene expression in mouse craniofacial bone development. AB - BACKGROUND: Annexin 1 is a 37-kDa protein that has complex intra- and extracellular effects. To discover whether the absence of this protein alters bone development, we monitored this event in the annexin-A1 null mice in comparison with littermate wild-type controls. METHODS: Radiographic and densitometry methods were used for the assessment of bone in annexin-A1 null mice at a gross level. We used whole-skeleton staining, histological analysis, and Western blotting techniques to monitor changes at the tissue and cellular levels. RESULTS: There were no gross differences in the appendicular skeleton between the genotypes, but an anomalous development of the skull was observed in the annexin A1 null mice. This was characterized in the newborn annexin-A1 null animals by a delayed intramembranous ossification of the skull, incomplete fusion of the interfrontal suture and palatine bone, and the presence of an abnormal suture structure. The annexin-A1 gene was shown to be active in osteocytes during this phase and COX-2 was abundantly expressed in cartilage and bone taken from annexin A1 null mice. CONCLUSIONS: Expression of the annexin-A1 gene is important for the normal development of the skull in mice, possibly through the regulation of osteoblast differentiation and a secondary effect on the expression of components of the cPLA2-COX-2 system. PMID- 17405165 TI - Active ammonia excretion in the giant mudskipper, Periophthalmodon schlosseri (Pallas), during emersion. AB - The main objective of this study was to determine whether active NH(4) (+) excretion occurred in the giant mudskipper, Periophthalmodon schlosseri, during emersion. Our results demonstrated that continual ammonia excretion in P. schlosseri during 24 hr of emersion resulted in high concentrations ( approximately 30 mmol l(-1)) of ammonia in fluid collected from the branchial surface. For fish injected intraperitoneally with 8 mumol g(-1) ammonium acetate (CH3COONH4) followed by 24 hr of emersion, the cumulative ammonia excreted was significantly greater than that of the control injected with sodium acetate. More importantly, the ammonia excretion rate at hour 2 in fish injected with CH3COONH4 followed by emersion was greater than that in fish immersed in water as reported elsewhere, with the greatest change in the ammonia excretion rate occurring at hour 2. Assuming that the rate of endogenous ammonia production remained unchanged, 33% of the exogenous ammonia was excreted through the head region, presumably through the gills, during the first 6 hr of emersion. Indeed, at hour 6, the ammonia concentration in the branchial fluid increased to an extraordinarily high concentration of >90 mmol l(-1). Therefore, our results confirm for the first time that P. schlosseri can effectively excrete a high load of ammonia on land, and corroborate the proposition that active NH(4) (+) excretion through its gills contributes in part to its high tolerance of aerial exposure. Only 4.6% of the exogenous ammonia was detoxified to urea. The glutamate contents in the muscle and liver also increased significantly, but the glutamine contents remained unchanged. PMID- 17405166 TI - Effects of unidirectional permeability in asymmetric poly(DL-lactic acid-co glycolic acid) conduits on peripheral nerve regeneration: an in vitro and in vivo study. AB - The high outflow permeability of the nerve conduit used to emit the drained waste generated from the traumatized host nerve stump is critical in peripheral nerve regeneration. Our earlier studies have established that asymmetric conduits fulfill the basic requirements for use as nerve guide conduits. In this study, the inflow characteristics of optimal nerve conduits were further examined using in vivo and in vitro trials. Various asymmetric poly(DL-lactic acid-co-glycolic acid) (PLGA) conduits were controlled by modifying precipitation baths using 0, 20, and 95% isopropyl alcohol, with high-porosity (permeability), medium-porosity (high outflow and low inflow), and low-porosity (permeability), respectively. In the in vitro trial, the Schwann cells and fibroblasts were seeded on either side of the asymmetric PLGA films in a newly designed coculture system that simulated the repaired nerve conduit environment. The results of the directional permeable films indicated the statistically significant proliferation of Schwann cells and the inhibition of the division of fibroblasts in lactate dehydrogenase release and inhibition of 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) reduction, compared with the other films. In the in vivo trial, the PLGA conduits seeded with Schwann cells were implanted into 10 mm right sciatic nerve defects in rats. After 6 weeks, implanted conduits were harvested. Histological examination verified that directional permeable conduits had markedly more A-type and B-type myelin fibers in the midconduit and distal nerve. In this work, the directional transport characteristics were established as an extremely important factor to the design and development of optimal nerve guide conduits in peripheral nerve regeneration. PMID- 17405167 TI - Structure and properties of methacrylate-endcapped caprolactone networks with modulated water uptake for biomedical applications. AB - Methacrylate-endcapped caprolactone (CLMA) networks were synthesized and copolymerized with 2-hydroxyethyl acrylate (HEA) seeking to tailor the hydrophilicity of the system. The resulting structure of the copolymer network is investigated by differential scanning calorimetry, thermogravimetry, and Fourier transform infrared spectroscopy. The dynamic swelling behavior and the equilibrium water sorption is measured and correlated with the microstructure. The experimental results allow one to conclude that the new material is a random copolymer of both components, HEA and CLMA. The effect of cell attachment and proliferation on the new copolymer networks was observed by in vitro culture of human chondrocytes up to 8 days. Enhanced cellular adhesion, similar to that obtained with tissue culture polystyrene (TCPS), was obtained in the hydrophilized systems. The new copolymers are appropriate for the fabrication of scaffolds with controlled porosity for tissue engineering. PMID- 17405168 TI - External validation of a risk group defined by recursive partitioning analysis in patients with head and neck carcinoma treated with surgery and postoperative radiotherapy. AB - BACKGROUND: Several clinical trials have proved that concurrent chemoradiotherapy is more efficacious than radiotherapy alone among high-risk patients with head and neck squamous cell carcinoma (HNSCC) who undergo surgery. A risk-group classification defined according to a recursive partitioning analysis (RPA) for these patients has been recently proposed. The objective of the present study was to carry out an external validation of this RPA-derived classification system. METHODS: A retrospective study of 442 HNSCC patients treated with surgery and postoperative radiotherapy was conducted. The external validity of the RPA derived classification system was assessed, and its ability to stage patients and to predict locoregional control of the disease was compared with the TNM system. RESULTS: The RPA-derived classification system succeeded in obtaining a monotonic prognosis gradient in locoregional control of the disease with increasing stage, and achieved greater differences in survival between stages than the TNM and pTNM classifications. Besides, the RPA method had a better homogeneity of the categories included in each stage, and in the heterogeneity between stages. CONCLUSIONS: The RPA-derived classification system allowed for the clear definition of prognostic groups in surgically treated HNSCC patients, improving the prognostic capacity of the TNM and pTNM classifications. The RPA-derived classification system is a useful tool in the definition of patients who, given a poor prognosis, should be considered candidates to adjuvant chemoradiotherapy. PMID- 17405169 TI - Changes in body mass, energy balance, physical function, and inflammatory state in patients with locally advanced head and neck cancer treated with concurrent chemoradiation after low-dose induction chemotherapy. AB - BACKGROUND: We aimed to determine changes in body mass and body composition in relation to energy balance, inflammatory state, and physical function before and after concurrent chemoradiation (CCR). METHODS: Seventeen patients with stage III and IVa head and neck cancer, aged 58.9 +/- 5.4 years, who had completed a 9-week regimen of low-dose induction chemotherapy came to the General Clinical Research Center pre- and post-CCR for measurement of body mass composition by dual-energy X-ray absorptiometry, resting energy expenditure (REE) by indirect calorimetry, physical performance (by Modified Baecke Questionnaire and Reuben's Physical Performance Test), and functionality (Activities and Instrumental Activities of Daily Living scores). Fasting venous samples were collected to determine C reactive protein and cytokines interleukin (IL)-1beta, IL-6, IL-8, and IL-10. Random 24-hour telephone diet recalls assessed energy intakes. RESULTS: Weight loss began 1 week after CCR. Lean body mass (LBM) accounted for 71.7% +/- 21% of body mass loss. No change occurred in energy intakes or calorie/nitrogen ratio. REE was significantly increased when adjusted for LBM loss (kcal/kg), p = .019. LBM loss was significantly associated with physical performance decline, r = .71, p = .004, and increased functional dependence, r = .58, p = .02. Total physical activity level declined significantly, p = .003. Cytokine levels were strongly associated with physical and functional decline. CONCLUSIONS: The aberrant changes in body composition, metabolism, and inflammatory state were associated with clinically and statistically significant impairments in physical performance and function. Future investigations and clinical practice should combine nutrition with antiinflammatory agents and exercise activities to support lean tissue anabolism and prevent physical and functional decline of patients with head and neck cancer undergoing CCR. PMID- 17405170 TI - Role of activated nuclear factor-kappaB in the pathogenesis and therapy of squamous cell carcinoma of the head and neck. AB - BACKGROUND: Nuclear factor-kappaB (NF-kappaB), a transcription factor known to modulate expression of factors involved in inflammation, immunity, proliferation, and apoptosis, is constitutively activated and plays a role in pathogenesis and therapeutic resistance in head and neck squamous cell carcinoma (HNSCC). Understanding the molecular alterations leading to aberrant NF-kappaB activation in HNSCC may direct investigators to novel therapeutic targets. METHODS: Results of laboratory and clinical studies are reviewed. RESULTS: The structure, function, and activation of NF-kappaB, products of NF-kappaB target genes and their role in HNSCC oncogenesis, and current NF-kappaB modulating interventions are described. CONCLUSIONS: Aberrant NF-kappaB activation contributes to the expression of oncogenes and the malignant phenotype observed in HNSCC. NF-kappaB, along with providing a link between cancer and inflammation, may serve as an appropriate therapeutic target to inhibit tumor growth and sensitize cancer cells to established cytotoxic anticancer therapies. PMID- 17405171 TI - Prospective randomized clinical trial of the effect of needle size on pain, sample adequacy and accuracy in head and neck fine-needle aspiration cytology. AB - BACKGROUND: Fine-needle aspiration cytology (FNAC) is useful in the diagnosis of many conditions of the head and neck. No reported studies have investigated the optimal needle gauge in the head and neck region. METHODS: This was a prospective randomized trial of 100 patients who required FNAC performed with either a 21G or 23G needle. Discomfort was scored by patients using a visual analogue scale. Sample accuracy was assessed in patients who subsequently had excision biopsy. RESULTS: FNAC performed with a 23G needle was less painful (mean +/- SE, 1.6 +/- 1.01) than FNAC with a 21G needle (3.3 +/- 1.94; p < .001). There was no difference in sample adequacy between the 2 needle sizes (p = .59). The sensitivities and specificities of both needles were similar. CONCLUSION: Head and neck FNAC should routinely be performed with a 23G needle, resulting in less patient discomfort, and giving sample adequacy comparable to a conventional 21G needle. PMID- 17405172 TI - Recurrent glottic cancer. PMID- 17405173 TI - Is central neck dissection necessary for the treatment of lateral cervical nodal recurrence of papillary thyroid carcinoma? AB - BACKGROUND: Although the pattern of cervical lymph node metastases from papillary thyroid carcinoma (PTC) has been described, little is known about the pattern of lateral cervical nodal recurrence. The aim of this study was to establish the optimal strategy for neck dissection in patients who underwent reoperation for lateral cervical recurrence of PTC. METHODS: We reviewed the records of 22 patients who underwent neck dissection for lateral nodal recurrence of thyroid cancer between 2002 and 2004. Eight patients had thyroid remnants or recurrent tumors in the bed and 6 had undergone lateral neck dissection prior to referral. Patients underwent comprehensive dissection of the posterolateral and ipsilateral (n = 10) or bilateral (n = 12) central neck. The pattern of nodal recurrence and postoperative morbidity were analyzed. RESULTS: All patients had lateral compartment involvement, 91% at mid-lower, 45% at upper, and 18% at posterior sites. Central nodes were involved in 86% of patients: 82% at ipsilateral paratracheal, 32% at pretracheal, 27% at superior mediastinal, and 2 patients at contralateral sites. Skip lateral recurrence with no positive central nodes was rarely observed (14%). Postoperative vocal cord palsy (n = 1) and hypoparathyroidism (n = 5) developed only in patients undergoing bilateral central compartment dissection. CONCLUSIONS: The inclusion of comprehensive ipsilateral central and lateral neck dissection in the reoperation for patients with lateral neck recurrence of PTC is an optimal surgical strategy. PMID- 17405174 TI - Functional outcomes after surgical reconstruction of the base of tongue using the radial forearm free flap in patients with oropharyngeal carcinoma. AB - BACKGROUND: Historically, the prevention of functional morbidity after multimodal treatment for squamous cell carcinoma in patients with large base of tongue lesions has been challenging. The purpose of the present research is to report prospectively collected speech and swallowing outcomes for patients with disease that encompassed half or more of the base of tongue. METHODS: Speech and swallowing data were gathered at 4 evaluation times that spanned the first year of treatment for 32 patients treated with surgery and adjuvant radiation therapy. RESULTS: Speech intelligibility remained above 90% for most patients. In the early postoperative period, patients with 100% of their base of tongue resected experienced decreased intelligibility, which improved by the half-way mark of the evaluations. The majority of patients resumed normal oral feeding, with those who lacked dentition needing to puree their food. Modified barium swallow studies revealed that only 3 patients aspirated thin liquid at the final evaluation. Five patients used a feeding tube at some point over the year of evaluation. At the final evaluation, only 3 patients required enteral feeding, with 2 of these using the tube in combination with oral feeding and 1 using the tube for primary nutrition. CONCLUSIONS: The results of this study suggest that surgical reconstruction of the base of tongue using the radial forearm free flap in patients with large base of tongue lesions can lead to functional speech and swallowing results. PMID- 17405175 TI - Molecular engineering of epoxide hydrolase and its application to asymmetric and enantioconvergent hydrolysis. AB - Safety and regulatory issues favor increasing use of enantiopure compounds in pharmaceuticals. Enantiopure epoxides and diols are valuable intermediates in organic synthesis for the production of optically active pharmaceuticals. Enantiopure epoxide can be prepared using epoxide hydrolase (EH)-catalyzed asymmetric hydrolysis of its racemate. Enantioconvergent hydrolysis of racemic epoxides by EHs possessing complementary enantioselectivity and regioselectivity can lead to the formation of enantiopure vicinal diols with high yield. EHs are cofactor-independent and easy-to-use catalysts. EHs will attract much attention as commercial biocatalysts for the preparation of enantiopure epoxides and diols. In this paper, recent progress in molecular engineering of EHs is reviewed. Some examples and prospects of asymmetric and enantioconvergent hydrolysis reactions are discussed as supplements to molecular engineering to improve EH performance. PMID- 17405176 TI - Multi-objective optimization of an industrial penicillin V bioreactor train using non-dominated sorting genetic algorithm. AB - Bulk of the penicillin produced is used as raw material for semi-synthetic penicillin (such as amoxicillin and ampicillin) and semi-synthetic cephalosporins (such as cephalexin and cefadroxil). In the present paper, an industrial penicillin V bioreactor train is optimized for multiple objectives simultaneously. An industrial train, comprising a bank of identical bioreactors, is run semi-continuously in a synchronous fashion. The fermentation taking place in a bioreactor is modeled using a morphologically structured mechanism. For multi-objective optimization for two and three objectives, the elitist non dominated sorting genetic algorithm (NSGA-II) is chosen. Instead of a single optimum as in the traditional optimization, a wide range of optimal design and operating conditions depicting trade-offs of key performance indicators such as batch cycle time, yield, profit and penicillin concentration, is successfully obtained. The effects of design and operating variables on the optimal solutions are discussed in detail. PMID- 17405177 TI - Genome-scale analysis of Mannheimia succiniciproducens metabolism. AB - Mannheimia succiniciproducens MBEL55E isolated from bovine rumen is a capnophilic gram-negative bacterium that efficiently produces succinic acid, an industrially important four carbon dicarboxylic acid. In order to design a metabolically engineered strain which is capable of producing succinic acid with high yield and productivity, it is essential to optimize the whole metabolism at the systems level. Consequently, in silico modeling and simulation of the genome-scale metabolic network was employed for genome-scale analysis and efficient design of metabolic engineering experiments. The genome-scale metabolic network of M. succiniciproducens consisting of 686 reactions and 519 metabolites was constructed based on reannotation and validation experiments. With the reconstructed model, the network structure and key metabolic characteristics allowing highly efficient production of succinic acid were deciphered; these include strong PEP carboxylation, branched TCA cycle, relative weak pyruvate formation, the lack of glyoxylate shunt, and non-PTS for glucose uptake. Constraints-based flux analyses were then carried out under various environmental and genetic conditions to validate the genome-scale metabolic model and to decipher the altered metabolic characteristics. Predictions based on constraints based flux analysis were mostly in excellent agreement with the experimental data. In silico knockout studies allowed prediction of new metabolic engineering strategies for the enhanced production of succinic acid. This genome-scale in silico model can serve as a platform for the systematic prediction of physiological responses of M. succiniciproducens to various environmental and genetic perturbations and consequently for designing rational strategies for strain improvement. PMID- 17405178 TI - Kinetics of microbial bromate reduction in a hydrogen-oxidizing, denitrifying biofilm reactor. AB - Bromate (BrO(3)(-)) is an oxidized contaminant produced from bromide (Br(-)) during ozonation and advanced oxidation of drinking water. Previous research shows that denitrifying bioreactors can reduce bromate to innocuous bromide. We studied a hydrogen-based, denitrifying membrane-biofilm reactor (MBfR) for bromate reduction, and report the first kinetics for a hydrogen-based bromate reduction process. A mixed-culture MBfR reduced up to 1,500 microg/L bromate to below 10 microg/L with a 50-min hydraulic residence time. Kinetics were determined using short-term tests on a completely mixed MBfR at steady state with an influent of 5 mg N/L nitrate plus 100 microg/L bromate. Short-term tests examined the impact of pH, nitrite, nitrate, and bromate on bromate reduction rates in the MBfR. Kinetic parameters for the process were estimated based on the short-term bromate tests. The q(max) for bromate reduction was 0.12 mg BrO(3)(-) x mg(x)(-1) x day(-1), and the K was 1.2 mg BrO(3)(-)/L. This q(max) is 2-3 times higher than reported for heterotrophic enrichments, and the K is the first reported in the literature. Nitrite and nitrate partially inhibited bromate reduction, with nitrite exerting a stronger inhibitory effect. Bromate was self inhibitory at concentrations above 15 mg/L, but up to 50 mg/L of bromate had no inhibitory effect on denitrification. The optimum pH was approximately 7. We also examined the performance of an MBfR containing pure culture of the denitrifying bacterium Ralstonia eutropha. Under conditions similar to the mixed-culture tests, no bromate reduction was detected, showing that not all denitrifying bacteria are active in bromate reduction. Our results suggest the presence of specialized, dissimilatory bromate-reducing bacteria in the mixed-culture MBfR. PMID- 17405179 TI - Development of a slow non-viral DNA release system from PDLLA scaffolds fabricated using a supercritical CO2 technique. AB - Polyamidoamine polymers (PAA) comprising methylene-bisacrylamide/dimethylethylene diamine monomers were synthesized, complexed with DNA and incorporated into porous P(DL)LA scaffolds by using a supercritical CO(2) (scCO(2)) technique. Scaffolds were made in a dry state consequently there was a need to lyophilize the complexes. A statistically significant reduction of the transfection efficiency was observed in the absence of trehalose when compared to the original complex after freeze-drying. Increasing concentrations (0-10% w/v) of trehalose were added to the complex prior to freeze-drying. Structure dependent differences in DNA binding were evaluated by gel electrophoresis and thermal transition analysis. TEM and PCS showed aggregate formation after freeze-drying without trehalose. Scaffolds were characterized by pore sizes of 173 +/- 73 microm and a porosity of 71%. The transfection potential of the released DNA was investigated by seeding scaffolds with A549 cells and following firefly luciferase as a marker gene after 48 h exposure. Low but continuous levels of transfection were observed for PAA complexes during a 60-day study. Complexes made with Lipofectaminetrade mark gave initially higher levels of DNA release but no further expression was seen after 40 days. Uncomplexed DNA showed background levels of transfection. Culturing cells on 3D scaffolds showed a benefit in retention of transfection activity with time compared to 2D controls. Transfection levels could be increased when cells were grown in OptiMEM. This study demonstrated that PAA/DNA complexes incorporated into a P(DL)LA scaffold made by using scCO(2) processing exhibited a slow release and extended gene expression profile. PMID- 17405180 TI - Characterization of antimicrobial histone sequences and posttranslational modifications by mass spectrometry. AB - Histones typically play a role in DNA packaging and transcription regulation. These proteins are heavily modified by acetylation, methylation, phosphorylation and/or ubiquitination, and various combinations of these modifications alter histone functions and form the basis of the histone code. Furthermore, histones, including those found in shrimp, have recently been found to possess antimicrobial properties; however, the sequences and posttranslational modifications of shrimp histones are largely unknown. In this study mass spectrometry was used to characterize the primary structure of the shrimp antimicrobial histone. A combination of in-solution digestion and in-gel propionylation/digestion followed by LC-MS-MS and MALDI-TOF-TOF analysis was used. Over 80% of each histone sequence was obtained by in-solution digestion; however, none of the N-terminal domains was sequenced with this method. An in-gel propionylation method was optimized to recover and sequence the extremely hydrophilic histone N-termini. This method was then applied to shrimp hemocyte lysates separated on a 1-D SDS-PAGE gel. Overall, 95% coverage was obtained for the histone sequences as well as the identification of posttranslational sites such as acetylation, methylation and phosphorylation. PMID- 17405182 TI - Structure-function analysis of protein complexes involved in the molecular architecture of juxtaparanodal regions of myelinated fibers. AB - Demyelinating disorders, including multiple sclerosis (MS), are common causes of neurological disability. One critical step towards the management and therapy of demyelinating diseases is to understand the basic functions of myelinating glia and their relationship with axons. Axons and myelinating glia, oligodendrocytes in the central (CNS) and Schwann cells in the peripheral (PNS) nervous systems, reciprocally influence each other's development and trophism. These interactions are critical for the formation of distinct axonal domains in myelinated fibers that ensure the rapid propagation of action potentials. Macromolecular complexes mediating axo-glial interactions in these domains have been identified, consisting of members of the immunoglobulin superfamily (IgSF) of adhesion molecules and the neurexin/NCP superfamily as well as other proteins. We have investigated the molecular details of axo-glial interactions in the juxtaparanodal region of myelinated fibers by utilizing domain-specific GFP constructs and immunoprecipitation assays on transfected cells. We have shown that the immunoglobulin domains of the IgSF member TAG-1/Cnt-2 are necessary and sufficient for the direct, cis interaction of this protein with Caspr2 and potassium channels. PMID- 17405184 TI - Evaluation of relative DNA binding affinities of anthrapyrazoles by electrospray ionization mass spectrometry. AB - Binding interactions of a new series of anthrapyrazoles (APs) with DNA were evaluated by electrospray ionization mass spectrometry (ESI-MS). Relative binding affinities were estimated from the ESI-MS data based on the fraction of bound DNA for DNA/anthrapyrazole mixtures, and they show a correlation to the shift in melting point of the DNA measured from a previous study. Minimal sequence specificity was observed for the series of anthrapyrazoles. Upon collisionally activated dissociation of the duplex/anthrapyrazole complexes, typically ejection of the ligand was the dominant pathway for most of the complexes. However, for complexes containing AP2 or mitoxantrone, strand separation with the ligand remaining on one of the single strands was observed, indicative of a different binding mode or stronger binding. PMID- 17405185 TI - Willingness-to-pay for a statistical life in the times of a pandemic. AB - A contingent valuation was performed based on cross-sectional web-based interviews of individuals aged 16-82 years of age presenting a scenario of influenza pandemic. The mean WTP for a course of Tamiflu was NOK 1034 (median NOK 250). Anxious individuals perceived the pandemic mortality risk to be higher than others, but also perceived the benefit of Tamiflu as greater. They also expressed a higher WTP for Tamiflu, but the implied VOSL was less than for other respondents. The results suggest that fear and anxiety distort decision making under uncertainty and decrease focus on the perceived risk reduction. PMID- 17405186 TI - Altered cryptal expression of luminal potassium (BK) channels in ulcerative colitis. AB - Decreased sodium (Na(+)), chloride (Cl(-)), and water absorption, and increased potassium (K(+)) secretion, contribute to the pathogenesis of diarrhoea in ulcerative colitis. The cellular abnormalities underlying decreased Na(+) and Cl( ) absorption are becoming clearer, but the mechanism of increased K(+) secretion is unknown. Human colon is normally a K(+) secretory epithelium, making it likely that K(+) channels are expressed in the luminal (apical) membrane. Based on the assumption that these K(+) channels resembled the high conductance luminal K(+) (BK) channels previously identified in rat colon, we used molecular and patch clamp recording techniques to evaluate BK channel expression in normal and inflamed human colon, and the distribution and characteristics of these channels in normal colon. In normal colon, BK channel alpha-subunit protein was immunolocalized to surface cells and upper crypt cells. By contrast, in ulcerative colitis, although BK channel alpha-subunit protein expression was unchanged in surface cells, it extended along the entire crypt irrespective of whether the disease was active or quiescent. BK channel alpha-subunit protein and mRNA expression (evaluated by western blotting and real-time PCR, respectively) were similar in the normal ascending and sigmoid colon. Of the four possible beta subunits (beta(1-4)), the beta(1)- and beta(3)-subunits were dominant. Voltage dependent, barium-inhibitable, luminal K(+) channels with a unitary conductance of 214 pS were identified at low abundance in the luminal membrane of surface cells around the openings of sigmoid colonic crypts. We conclude that increased faecal K(+) losses in ulcerative colitis, and possibly other diseases associated with altered colonic K(+) transport, may reflect wider expression of luminal BK channels along the crypt axis. PMID- 17405187 TI - Hepatocyte growth factor and c-Met expression in pericytes: implications for atherosclerotic plaque development. AB - Intraplaque neovascularization contributes to the progression of atherosclerosis. Our aim is to understand the mobilization of cells and factors involved in this process. We investigated the localization of hepatocyte growth factor (HGF) and its receptor, c-Met, in human atherosclerotic plaques, together with the effects of HGF on pericyte migration in vitro. Atherosclerotic femoral arterial segments were collected and analysed from 13 subjects who were undergoing lower limb amputation. Pericytes were identified in human lesions using a 3G5 antibody. Immunohistochemical analysis localized HGF mainly around microvessels, in association with some, but not all, CD31-positive endothelial cells. c-Met expression was mainly associated with smooth muscle cells and pericytes, around some, but not all, microvessels within the atherosclerotic lesions; no detection was apparent in normal internal mammary arteries. Using RT-PCR, we demonstrated expression of HGF and c-Met in a rat pericyte cell-line, TR-PCT1, and in primary pericytes. HGF treatment of TR-PCT1 cells induced their migration, but not their proliferation, in a dose-dependent manner (10-100 ng/ml, p<0.01), an effect mediated by activation of the serine/threonine kinase Akt, shown by western blot analysis. Treating the cells with the PI3K inhibitors Wortmannin (0.1 microM) or LY294002 (10 microM) abolished these effects. This work demonstrates the expression of c-Met and HGF in human atherosclerotic arteries, in association with SM-actin-positive cells and CD-31-positive cells, respectively. HGF induces pericyte migration via PI3-kinase and Akt activation in vitro. HGF and c-Met may be involved in neovascularization during plaque development, and may recruit pericytes to neovessels. Since pericytes are thought to mechanically stabilize new blood vessels, these factors may function to protect against haemorrhage. PMID- 17405188 TI - Refinement of cardiac NMR imaging in awake hamsters: proof of feasibility and characterization of cardiomyopathy. AB - The goal of this study was to demonstrate the feasibility of cardiac NMR imaging in conscious hamsters and its usefulness in evaluating cardiac abnormalities in a small-animal model of cardiomyopathy. Awake hamsters, controls and cardiomyopathic ones (CHF 147), were immobilized in a dedicated holder. Half Fourier single-shot FSE imaging, with outer-volume suppression and 'black-blood' contrast provided images free from motion artifact with good visualization of cardiac anatomy at any point in the cardiac cycle. Series of double-oblique views were acquired with or without electrocardiograph gating. Image acquisition time was 55 ms, with an in-plane resolution of 470 x 625 microm2. Left ventricular volumes, ejection fraction, and myocardium NMR signal heterogeneity were compared in CHF 147 and control hearts. Left ventricles of CHF 147 hamsters were dilated, as indicated by the increase in end-diastolic cavity volume (299 +/- 79 mm3 compared with the controls (141 +/- 39 mm3; P = 0.0002). Left ventricular ejection fraction was largely reduced (45 +/- 9% vs 86 +/- 4%; P < 0.0001). The NMR signal distribution at an effective echo time of 41 ms was more heterogeneous in the myocardial wall of CHF 147 hamsters than in controls (1.87 +/- 0.37 a.u. vs 0.98 +/- 0.12 a.u., respectively; P = 0.0002). This study is a refinement of animal experimentation, as it demonstrates for the first time that characteristic features of cardiac pathology can be evaluated with ultra-fast NMR imaging in conscious small rodents. PMID- 17405189 TI - Extremely high prevalence of DNASE1*1 allele in African populations. AB - The single nucleotide polymorphism (SNP) at deoxyribonuclease I (DNase I), designated as DNASE1 (NCBI SNP number; 1053874), in exon 8 (A2317G) has been shown to be associated with liver disease, colorectal carcinoma, and gastric carcinoma in Japanese patients. In this study, we investigated the frequency of the DNASE1 polymorphism in Ghanaian (n = 96) and Xhosa (n = 78) populations and compared the results with those of other studies. The single nucleotide polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The frequencies of DNASE1*1 in the Ghanaian and Xhosa populations were 0.90 and 0.88, respectively. These two African populations had an extremely high frequency of DNASE1*1, similar to that of the Ovambos living in Namibia. Caucasians and Asians had a lower frequency of DNASE1*1 than the African groups. This study is the first to reveal an extremely high frequency of DNASE1*1 among African populations. PMID- 17405190 TI - Very high-resolution three-dimensional functional MRI of the human visual cortex with elimination of large venous vessels. AB - We propose a very high-resolution, three-dimensional (3D) gradient-echo technique with a twofold parallel imaging acceleration using a specialized occipital receiver coil at 3 T to perform functional MRI (fMRI) of the visual cortex. This configuration makes it possible to acquire 3D fMRI data within a timescale compatible with a block design. Without further processing, the functional maps at an isotropic 3D resolution of 0.42 microL (0.75 mm voxel size) and near isotropic resolution of 1.2 microL (1 mm voxel size) show very robust activation in visual areas, but with clear contamination from larger veins. As this technique allows direct identification of veins in the functional scan, it permits removal of their effect from the activation maps. In our study, elimination of veins qualitatively improves the spatial specificity of activation maps, while reducing the activated volume by about 25%. The proposed technique provides functional information at the resolution of anatomical scans, is localized to gray matter, and facilitates functional to anatomical co registration because of minimal distortions. PMID- 17405191 TI - Informed consent for extensor indicis transfer. PMID- 17405192 TI - Evaluation of quality of life in brachial plexus injury patients after reconstructive surgery. AB - To evaluate the subjective satisfaction of brachial plexus injury (BPI) patients after surgery based on the medical outcomes study 36-item short form health survey (SF-36) and to correlate their SF-36 scores with upper extremity functions. Four items were assessed statistically for 30 patients: SF-36 scores after BPI surgery were compared with Japanese standard scores; the correlation between SF-36 scores and objective joint functions; difference in SF-36 scores between each type of BPI; and influence of each joint function on the SF-36 scores. The SF-36 subscale: PF--physical functioning, RP--role-physical, BP- bodily pain, and the summary score PCS--physical component summary, were significantly inferior to the Japanese standard scores. SF-36 is more sensitive to shoulder joint function than to elbow and finger joint functions. Little correlation was found between SF-36 scores and objective evaluations of joint functions. Greater effort is needed to improve the quality of life (QOL) of BPI patients. This study showed that SF-36 is not sensitive enough to evaluate regional conditions. A region- or site-specific questionnaire is required to evaluate upper extremity surgery. PMID- 17405193 TI - The early history of hand surgery in the Philippines and highlights in my experience at the National Orthopaedic Hospital Mandaluyong and the Our Lady of Lourdes Hospital, Manila. AB - Several years ago, while attending a Philippines Orthopaedic Association (PDA) Annual Convention, held at the Westin Philippine Plaza Hotel, Manila in December, I had the opportunity to meet with Professor Yoshikazu Ikuta, a well-known microsurgery and hand surgeon from Japan and one of the foreign guest speakers. I had been invited to work with him at the Armed Forces of the Philippines (AFP) Medical Center on the case of a high ranking general of the AFP, with a hand contracture disability problem. The case was referred to him by Dr. Evaristo Sanchez, Chief of Orthopaedics and the Commanding General of the AFP Medical Center. He had been pre-scheduled for surgery the next day, a Sunday morning, the day before Prof. Ikuta was due to return to Japan. After a brief examination and evaluation of the generals affected hand, in the operating room with Prof. Ikuta, just before he was placed under general anaesthesia, we performed the operation together. The operation did not involve microsurgery. The procedures done were multiple combined Bunnell-Zancolli pulley advancements and MP-joint volar capsulorraphies plus flexor tendon releases in the volar forearm, which although quite extensive, were only palliative, to minimise and improve on the contracture deformities, in preparation for a final re-evaluation for possible later, more definitive tendon transfers for hand function. However, I never received any further information regarding the results of our surgery. Recently, I have been honoured and invited again by Prof. Ikuta, presently the Editor-in-Chief of the Hand Surgery Journal (Asian Volume), this time to write the history of hand surgery in the Philippines and add to it, "Highlights in my experience at the National Orthopaedic Hospital, Mandaluyong, and the Our Lady of Lourdes Hospital, Manila", the last portion of which is on paralytic disabilities of the Hand. I am deeply grateful to Prof. Ikuta for giving me this honour and opportunity to present the total experience, favourable and unfavourable, of a hand surgeon from a developing country, like the Philippines. Furthermore, this would afford me also, the chance to be able to make known to readers of this now prestigious journal, the philosophical thoughts which led me to unwittingly originate or come up with and develop a few of my own "Long Tendon Rerouting Procedures" which may possibly and hopefully merit as this author's title contribution to surgery of the hands. PMID- 17405194 TI - The association of scapho-trapezio-trapezoid osteoarthrosis and scapholunate dissociation. AB - There are two parts to this paper. In the first part we report 18 cases of bilateral scapho-trapezio-trapezoid osteoarthrosis associated with uni- or bilateral scapholunate dissociation. This case series was followed prospectively using repeat clinical assessments and radiographs. We were able to document the progression of both the clinical manifestations and radiographic features of the scapholunate dissociation in these patients. In the second part of the paper we compared the radiographic indices of scapholunate dissociation seen in our series, that is, the scapholunate angle and interval, with those of a control group over time, to determine if there was a significant difference. The control group differed from our series principally by being devoid of any clinical or radiographic evidence of scapho-trapezio-trapezoid osteoarthrosis. We found that our case series already had some radiographic evidence of scapholunate attrition at presentation and that at follow-up the scapholunate dissociation became more pronounced both clinically and radiographically relative to the controls. The authors propose a theory to explain the association and temporal relationship between the two conditions. PMID- 17405195 TI - Osteochondral destruction in the hand following bee stings: a case report of an infant. AB - We report an infant who developed arthritis with osteochondral destruction in a hand after being stung by bees. The clinical findings resembled those reported as "beekeeper's arthropathy", but the radiological findings were more severe. Histology of the resected specimen revealed fibrous granulation tissue with infiltration of inflammation cells. PMID- 17405196 TI - Acute bilateral forearm compartment syndrome of unknown aetiology. AB - This is the first reported case of non-traumatic, acute bilateral forearm compartment syndrome. Despite a delay of over 24 hours until surgical decompression and 50% muscle fibre necrosis in the histopathological examination, the clinical outcome was excellent after fasciotomy, delayed primary wound closure and early institution of a range of motion exercise programme. The literature on non-traumatic causes of compartment syndrome is reviewed. PMID- 17405197 TI - Scaphoid non-union after simultaneous fractures of the scaphoid and the capitate in a child: a case report. AB - A nine-year-old boy fell on his outstretched hand and was diagnosed with simultaneous fractures of the scaphoid and the capitate. The patient was treated conservatively with a cast for two months. Fracture of the capitate healed, however, fracture of the scaphoid resulted in non-union. We treated scaphoid non union with an iliac bone graft and internal fixation. Three years after surgery, the patient had neither complaints nor complications and union of the scaphoid was confirmed on the radiographs. PMID- 17405198 TI - Minimally invasive technique for finger flexor pulley reconstruction. AB - A minimal invasive technique for finger flexor pulley is described. The technique is compared biomechanically to a "single loop" tendon graft technique using a cadaveric model. PMID- 17405199 TI - Transdermal anaesthesia for percutaneous trigger finger release. AB - The purpose of this study was to evaluate the safety and efficiency of transdermal anaesthesia using eutectic mixture of lidocaine and prilocaine (EMLA) in patients undergoing percutaneous trigger finger release and to compare it with lidocaine infiltration. In this prospective, randomised study percutaneous release of the A1 annular pulley was performed to treat stenosing tenosynovitis (trigger finger syndrome) in 50 patients (50 fingers). The procedure was performed either under transdermal anaesthesia using EMLA applied transcutaneously 120 minutes prior to the operation (Group A, n = 25) or using local infiltration anaesthesia using lidocaine (Group B, n = 25). Pain experienced during administration of anaesthesia and during the operation was assessed using a 10-point Visual Analogue Pain Scale (VAPS), while all patients rated the effectiveness of anaesthesia with a 5-point scale. There were no significant differences between the two groups in the VAPS during the operation (1.33 +/- 0.52 versus 1.59 +/- 0.87) and the satisfaction scores (4.6 +/- 0.2 versus 4.4 +/- 0.3). The VAPS score during the administration of anaesthesia was statistically significantly less in the EMLA group (0 versus 5.96 +/- 2.41). All patients were satisfied with the final result of the operation. Percutaneous trigger finger release can be performed as an office procedure with the use of EMLA avoiding the use of injectable local infiltration anaesthesia. PMID- 17405200 TI - Africa battles to make female genital mutilation history. PMID- 17405201 TI - US scientists press Congress to boost NIH funding. PMID- 17405202 TI - Report slams US FDA's drug safety reform efforts. PMID- 17405203 TI - Chemistry in context. PMID- 17405204 TI - Researchers without frontiers. PMID- 17405205 TI - Antarctic ecology from genes to ecosystems: the impact of climate change and the importance of scale. AB - Antarctica offers a unique natural laboratory for undertaking fundamental research on the relationship between climate, evolutionary processes and molecular adaptation. The fragmentation of Gondwana and the development of wide scale glaciation have resulted in major episodes of extinction and vicariance, as well as driving adaptation to an extreme environment. On shorter time-scales, glacial cycles have resulted in shifts in distribution, range fragmentation and allopatric speciation, and the Antarctic Peninsula is currently experiencing among the most rapid climatic warming on the planet. The recent revolution in molecular techniques has provided a suite of innovative and powerful tools to explore the consequences of these changes, and these are now providing novel insights into evolutionary and ecological processes in Antarctica. In addition, the increasing use of remotely sensed data is providing a large-scale view of the system that allows these processes to be set in a wider spatial context. In these two volumes, we collect a wide range of papers exploring these themes, concentrating on recent advances and emphasizing the importance of spatial and temporal scale in understanding ecological and evolutionary processes in Antarctica. PMID- 17405206 TI - Disturbance, colonization and development of Antarctic benthic communities. AB - A decade has yielded much progress in understanding polar disturbance and community recovery-mainly through quantifying ice scour rates, other disturbance levels, larval abundance and diversity, colonization rates and response of benthos to predicted climate change. The continental shelf around Antarctica is clearly subject to massive disturbance, but remarkably across so many scales. In summer, millions of icebergs from sizes smaller than cars to larger than countries ground out and gouge the sea floor and crush the benthic communities there, while the highest wind speeds create the highest waves to pound the coast. In winter, the calm associated with the sea surface freezing creates the clearest marine water in the world. But in winter, an ice foot encases coastal life and anchor ice rips benthos from the sea floor. Over tens and hundreds of thousands of years, glaciations have done the same on continental scales-ice sheets have bulldozed the seabed and the zoobenthos to edge of shelves. We detail and rank modern disturbance levels (from most to least): ice; asteroid impacts; sediment instability; wind/wave action; pollution; UV irradiation; volcanism; trawling; non-indigenous species; freshwater inundation; and temperature stress. Benthic organisms have had to recolonize local scourings and continental shelves repeatedly, yet a decade of studies have demonstrated that they have (compared with lower latitudes) slow tempos of reproduction, colonization and growth. Despite massive disturbance levels and slow recolonization potential, the Antarctic shelf has a much richer fauna than would be expected for its area. Now, West Antarctica is among the fastest warming regions and its organisms face new rapid changes. In the next century, temperature stress and non-indigenous species will drastically rise to become dominant disturbances to the Antarctic life. Here, we describe the potential for benthic organisms to respond to disturbance, focusing particularly on what we know now that we did not a decade ago. PMID- 17405207 TI - The biodiversity of the deep Southern Ocean benthos. AB - Our knowledge of the biodiversity of the Southern Ocean (SO) deep benthos is scarce. In this review, we describe the general biodiversity patterns of meio-, macro- and megafaunal taxa, based on historical and recent expeditions, and against the background of the geological events and phylogenetic relationships that have influenced the biodiversity and evolution of the investigated taxa. The relationship of the fauna to environmental parameters, such as water depth, sediment type, food availability and carbonate solubility, as well as species interrelationships, probably have shaped present-day biodiversity patterns as much as evolution. However, different taxa exhibit different large-scale biodiversity and biogeographic patterns. Moreover, there is rarely any clear relationship of biodiversity pattern with depth, latitude or environmental parameters, such as sediment composition or grain size. Similarities and differences between the SO biodiversity and biodiversity of global oceans are outlined. The high percentage (often more than 90%) of new species in almost all taxa, as well as the high degree of endemism of many groups, may reflect undersampling of the area, and it is likely to decrease as more information is gathered about SO deep-sea biodiversity by future expeditions. Indeed, among certain taxa such as the Foraminifera, close links at the species level are already apparent between deep Weddell Sea faunas and those from similar depths in the North Atlantic and Arctic. With regard to the vertical zonation from the shelf edge into deep water, biodiversity patterns among some taxa in the SO might differ from those in other deep-sea areas, due to the deep Antarctic shelf and the evolution of eurybathy in many species, as well as to deep-water production that can fuel the SO deep sea with freshly produced organic matter derived not only from phytoplankton, but also from ice algae. PMID- 17405208 TI - Marine pelagic ecosystems: the west Antarctic Peninsula. AB - The marine ecosystem of the West Antarctic Peninsula (WAP) extends from the Bellingshausen Sea to the northern tip of the peninsula and from the mostly glaciated coast across the continental shelf to the shelf break in the west. The glacially sculpted coastline along the peninsula is highly convoluted and characterized by deep embayments that are often interconnected by channels that facilitate transport of heat and nutrients into the shelf domain. The ecosystem is divided into three subregions, the continental slope, shelf and coastal regions, each with unique ocean dynamics, water mass and biological distributions. The WAP shelf lies within the Antarctic Sea Ice Zone (SIZ) and like other SIZs, the WAP system is very productive, supporting large stocks of marine mammals, birds and the Antarctic krill, Euphausia superba. Ecosystem dynamics is dominated by the seasonal and interannual variation in sea ice extent and retreat. The Antarctic Peninsula is one among the most rapidly warming regions on Earth, having experienced a 2 degrees C increase in the annual mean temperature and a 6 degrees C rise in the mean winter temperature since 1950. Delivery of heat from the Antarctic Circumpolar Current has increased significantly in the past decade, sufficient to drive to a 0.6 degrees C warming of the upper 300 m of shelf water. In the past 50 years and continuing in the twenty-first century, the warm, moist maritime climate of the northern WAP has been migrating south, displacing the once dominant cold, dry continental Antarctic climate and causing multi-level responses in the marine ecosystem. Ecosystem responses to the regional warming include increased heat transport, decreased sea ice extent and duration, local declines in icedependent Adelie penguins, increase in ice-tolerant gentoo and chinstrap penguins, alterations in phytoplankton and zooplankton community composition and changes in krill recruitment, abundance and availability to predators. The climate/ecological gradients extending along the WAP and the presence of monitoring systems, field stations and long-term research programmes make the region an invaluable observatory of climate change and marine ecosystem response. PMID- 17405209 TI - Trophic interactions within the Ross Sea continental shelf ecosystem. AB - The continental shelf of the Ross Sea is one of the Antarctic's most intensively studied regions. We review the available data on the region's physical characteristics (currents and ice concentrations) and their spatial variations, as well as components of the neritic food web, including lower and middle levels (phytoplankton, zooplankton, krill, fishes), the upper trophic levels (seals, penguins, pelagic birds, whales) and benthic fauna. A hypothetical food web is presented. Biotic interactions, such as the role of Euphausia crystallorophias and Pleuragramma antarcticum as grazers of lower levels and food for higher trophic levels, are suggested as being critical. The neritic food web contrasts dramatically with others in the Antarctic that appear to be structured around the keystone species Euphausia superba. Similarly, we suggest that benthic-pelagic coupling is stronger in the Ross Sea than in most other Antarctic regions. We also highlight many of the unknowns within the food web, and discuss the impacts of a changing Ross Sea habitat on the ecosystem. PMID- 17405210 TI - Spatial and temporal operation of the Scotia Sea ecosystem: a review of large scale links in a krill centred food web. AB - The Scotia Sea ecosystem is a major component of the circumpolar Southern Ocean system, where productivity and predator demand for prey are high. The eastward flowing Antarctic Circumpolar Current (ACC) and waters from the Weddell-Scotia Confluence dominate the physics of the Scotia Sea, leading to a strong advective flow, intense eddy activity and mixing. There is also strong seasonality, manifest by the changing irradiance and sea ice cover, which leads to shorter summers in the south. Summer phytoplankton blooms, which at times can cover an area of more than 0.5 million km2, probably result from the mixing of micronutrients into surface waters through the flow of the ACC over the Scotia Arc. This production is consumed by a range of species including Antarctic krill, which are the major prey item of large seabird and marine mammal populations. The flow of the ACC is steered north by the Scotia Arc, pushing polar water to lower latitudes, carrying with it krill during spring and summer, which subsidize food webs around South Georgia and the northern Scotia Arc. There is also marked interannual variability in winter sea ice distribution and sea surface temperatures that is linked to southern hemisphere-scale climate processes such as the El Nino-Southern Oscillation. This variation affects regional primary and secondary production and influences biogeochemical cycles. It also affects krill population dynamics and dispersal, which in turn impacts higher trophic level predator foraging, breeding performance and population dynamics. The ecosystem has also been highly perturbed as a result of harvesting over the last two centuries and significant ecological changes have also occurred in response to rapid regional warming during the second half of the twentieth century. This combination of historical perturbation and rapid regional change highlights that the Scotia Sea ecosystem is likely to show significant change over the next two to three decades, which may result in major ecological shifts. PMID- 17405211 TI - Climate change and the marine ecosystem of the western Antarctic Peninsula. AB - The Antarctic Peninsula is experiencing one of the fastest rates of regional climate change on Earth, resulting in the collapse of ice shelves, the retreat of glaciers and the exposure of new terrestrial habitat. In the nearby oceanic system, winter sea ice in the Bellingshausen and Amundsen seas has decreased in extent by 10% per decade, and shortened in seasonal duration. Surface waters have warmed by more than 1 K since the 1950s, and the Circumpolar Deep Water (CDW) of the Antarctic Circumpolar Current has also warmed. Of the changes observed in the marine ecosystem of the western Antarctic Peninsula (WAP) region to date, alterations in winter sea ice dynamics are the most likely to have had a direct impact on the marine fauna, principally through shifts in the extent and timing of habitat for ice-associated biota. Warming of seawater at depths below ca 100 m has yet to reach the levels that are biologically significant. Continued warming, or a change in the frequency of the flooding of CDW onto the WAP continental shelf may, however, induce sublethal effects that influence ecological interactions and hence food-web operation. The best evidence for recent changes in the ecosystem may come from organisms which record aspects of their population dynamics in their skeleton (such as molluscs or brachiopods) or where ecological interactions are preserved (such as in encrusting biota of hard substrata). In addition, a southwards shift of marine isotherms may induce a parallel migration of some taxa similar to that observed on land. The complexity of the Southern Ocean food web and the nonlinear nature of many interactions mean that predictions based on short-term studies of a small number of species are likely to be misleading. PMID- 17405212 TI - [Response to the open letter of the President-Elect of the European Society of Cardiology Roberto Ferrari, published in Il Giornale Italiano di Cardiologiia 1, 2007]. PMID- 17405213 TI - [Response to the open letter of the President-Elect of the European Society of Cardiology Roberto Ferrari, published in Il Giornale Italiano di Cardiologia 1, 2007]. PMID- 17405214 TI - [Proximal femur fractures in the elderly in The Netherlands during the period 1991-2004: incidence, mortality, length of hospital stay and an estimate of the care capacity needed in the future]. PMID- 17405215 TI - New approach to mental health: interview with Jeff Hoeltzel, Community Living Room. Interview by John S. James. AB - A Philadelphia program for people with a mental health diagnosis who are HIV positive has won national recognition. We interviewed its founder for ideas and approaches others can use. PMID- 17405216 TI - Fuzeon: avoiding injection-site reactions. AB - The FDA changed the FUZEON prescribing information to help avoid this common and unpleasant side effect of the drug. PMID- 17405217 TI - Standing under the arch. PMID- 17405218 TI - Micronutrient deficiencies less common in patients on ART. PMID- 17405219 TI - HAART, hubris, and humility. Decade of HAART. September 25-26, 2006--San Francisco. PMID- 17405220 TI - Focus on hepatitis. Weight-based RBV dosing = impressive treatment success rates. PMID- 17405221 TI - Darunavir offers potent new combo when used with enfuvirtide in ritonavir-boosted regimen. PMID- 17405222 TI - Telemedicine improves access to care for HIV-infected prisoners. PMID- 17405223 TI - State laws v. CDC: the future of HIV testing and informed consent. PMID- 17405224 TI - Nursing. In clinic, every team member counts when it comes to adherence. PMID- 17405225 TI - Periodontal disease is more aggressive in HIV-infected patients. PMID- 17405226 TI - How can patients protect themselves against foodborne illness? PMID- 17405227 TI - XAS and XPS characterization of mercury binding on brominated activated carbon. AB - Brominated powdered activated carbon sorbents have been shown to be quite effective for mercury capture when injected into the flue gas duct at coal-fired power plants and are especially useful when burning Western low-chlorine subbituminous coals. X-ray absorption spectroscopy (XAS) and X-ray photoelectron spectroscopy (XPS) have been used to determine information about the speciation and binding of mercury on two commercially available brominated activated carbons. The results are compared with similar analysis of a conventional (non halogenated) and chlorinated activated carbon. Both the XAS and XPS results indicate that the mercury, though introduced as elemental vapor, is consistently bound on the carbon in the oxidized form. The conventional and chlorinated activated carbons appeared to contain mercury bound to chlorinated sites and possibly to sulfate species that have been incorporated onto the carbon from adsorbed SO2. The mercury-containing brominated sorbents appear to contain mercury bound primarily at bromination sites. The mechanism of capture for the sorbents likely consists of surface-enhanced oxidation of the elemental mercury vapor via interaction with surface-bound halide species with subsequent binding by surface halide or sulfate species. PMID- 17405228 TI - Dining at the periodic table: metals concentrations as they relate to recycling. AB - A correlation between the prices of a variety of substances and their dilutions in their initial matrices was shown in 1959 by T.K. Sherwood. The research presented here shows that the relationship holds for engineering metals today, which we termed the metals-specific Sherwood plot. The concentrations of metals in products (e.g., printed wiring boards and automobiles) and waste streams (e.g., municipal solid waste, and construction and demolition debris) were plotted with this correlation. In addition, for the products and waste streams that undergo disassembly at end-of-life, the metals concentrations of the disassembled components were also plotted. It was found that most of the metals that are currently targeted for recycling have post-disassembly concentrations that lie above the metals-specific Sherwood plot (i.e., have concentrations that are more enriched than minimum profitable ore grades). This suggests that material concentration plays a role in the viability of recycling at end-of-life. As products grow in complexity and the variety of materials used, analyses such as this one provide insight for policymakers and those interested in material sustainability into macro-level trends of material use and future recycling practices. PMID- 17405229 TI - Batch synthesis of DNA. PMID- 17405230 TI - Reducing the health risks for children from ozone layer depletion: new ozone education pack targets primary schools. PMID- 17405231 TI - WHO gives indoor use of DDT a clean bill of health for controlling malaria. PMID- 17405232 TI - CDC recommends routine, voluntary HIV screening in health care settings. PMID- 17405233 TI - Introduction to anxiety sensitivity: recent findings and new directions. AB - There has been a long-standing recognition that cognitive factors may play a formative role in anxiety and its disorders (Barlow, 2002). Since the 1980s, there has been great progress made in understanding the nature of cognitive risk for anxiety psychopathology. Within this context, numerous cognitive factors have been implicated and studied as potential risk factors for various anxiety conditions. Perhaps one of the most well-known and promising of these has been anxiety sensitivity (AS; Reiss & McNally,1985). AS is the fear of anxiety-related bodily sensations, which arise from beliefs that the sensations have harmful personal consequences (McNally,2002). To illustrate, people high in AS may be frightened of harmless heart palpitations because they believe the sensations will lead to cardiac arrest,whereas people low in AS do not fear these sensations because they believe them to be harmless. High AS is theorized to enhance anxious and fearful responding to internal cues through associative learning and/or cognitive misinterpretation (McNally, 2002). Numerous studies have indicated that AS is, in fact, related to increased risk of panic attacks (e.g., Hayward, Killen, Kraemer, & Taylor, 2000; Schmidt, Lerew, & Jackson, 1997, 1999) and anxiety disorders (Schmidt, Zvolensky, & Maner, 2006). Although the vast majority of work on AS has been conducted in relation to panic psychopathology (Zvolensky, Schmidt, Bernstein, & Keough, 2006), researchers PMID- 17405234 TI - Prescriptions for sustainable healthcare. PMID- 17405236 TI - An up side to ozone depletion. PMID- 17405235 TI - [Treatment of acute bacterial tonsillopharyngitis in pediatrics: a meta analysis]. AB - Pharyngo-tonsillitis represents the most common infection of the upper respiratory tract, its treatment being the most common cause for prescribing antibiotics. Efficacy, safety and compliance of cefaclor were compared with those of other antibiotics in the treatment of paediatric acute bacterial tonsillo pharyngitis in a meta-analysis of randomized controlled trials published between 1979 and 2003. Overall, evaluations were performed on 16 studies (Medline/PubMed, keywords "Cefaclor and tonsillo-pharyngitis) which proved eligible (Jadad score > or = 1); twelve out of 16 studies were multicentre ones, only one was a double blind study. Mostly, the comparator agent was a beta-lactam, in four cases it was a macrolide. Efficacy and safety were end-points of all studies whereas only 13 and 4 studies evaluated adverse events and compliance, respectively. The analysis was based on a 2 x 2 contingency table with classification by treatment and number of improvements/cures, side-effects, and compliance of the individual studies. The global estimate of the effective treatment was obtained with the weighted mean of the log OR (Odds Ratio) according to Mantel-Haenszel and associated confidence intervals (CI) at 95%. Chi-square test was performed. All the calculations were performed using SAS v.8. Clinical efficacy evaluation, number of improvements/cures, did not evidence a statistically significant difference among cefaclor and comparators (93.8% vs 92.3%; Odds Ratio 1.21, IC 0.95/1.48). In the cefaclor-treated patients, adverse events were observed in a statistically significant lower percentage compared to other antibiotics: 8.5% vs 15.5% (Odds Ratio 0.49, IC 0.22/0.76; P < 0.0001). Compliance was observed in a similar proportion in both the two groups, cefaclor and comparators (Cefaclor, mean 100%; comparators, mean 98.3%). The present meta-analysis proves that in the treatment of paediatric acute bacterial tonsillo-pharyngitis cefaclor exhibits a clinical efficacy equal to other antibiotics usually employed in this setting, similar compliance but superior safety. PMID- 17405237 TI - Theoretically speaking. Interview by Hilary Crichton. PMID- 17405238 TI - Delivering the goods. PMID- 17405239 TI - [In vitro activity of prulifloxacin, levofloxacin and ciprofloxacin against urinary pathogens]. AB - The present investigation aimed to compare the in vitro antimicrobial activity of prulifloxacin to that of levofloxacin and ciprofloxacin against recently urinary pathogens. From our large bacterial collection, 978 non-duplicated clinical isolates, derived from patients urinary infections, maintained with minimal sub culture deep frozen at -70 degrees C in a matrix of Brain Heart Infusion broth +20 % glycerol were tested to determine MICs of prulifloxacin, levolfoxacin and ciprofloxacin by means of broth microdilution technique. For Gram negartive bacteria, no significant diffrences was observed between all three antibiotics. Enterobacteria showed a resistance rate ranging between 6% (Enterobacter spp) and approximately 20% (Escherichia coli). Antibiotic resistance of Pseudomas spp was higher (30.3%). For Gram positive, all methicillin-susceptible Stalphylococcus aureus strains were sensitive whereas 30% of methicillin-resistant and enterococci were resistant to all three antibiotics. Purification exerts its antibacterial activity at lower concentrations than ciprofloxacin and levofloxacin. The in vitro activity of prulifloxacin against urinary pathogens and its high and prolonged urinary concentration following a single oral dose, suggest that for treatment of urinary prulifloxacin is an alternative to other flouroquinolones for treatment of urinary tract infections. PMID- 17405240 TI - The impact of the introduction of premiums into a SCHIP program. AB - This paper examines the introduction of premiums into the SCHIP program in Kentucky. Kentucky introduced a $20 monthly premium for SCHIP coverage for children with family incomes between 151 percent and 200 percent of the federal poverty level in December 2003. Administrative data between 2001 and 2004 is used to estimate a Cox proportional hazard model that predicts enrollment duration in this premium-paying category. The results suggest that a premium reduces the length of enrollment, with the impact concentrated in the first three months after the introduction of the premium. Similar results are not found for the non premium-paying category. PMID- 17405241 TI - [Adverse effects of glitazones]. PMID- 17405242 TI - Consensus statement on self-monitoring in diabetes: Institute of Health Economics, Alberta, Canada, November 14-16, 2006. PMID- 17405243 TI - Regulating life and death: the case of Israel's "Health Basket" Committee. PMID- 17405244 TI - Cross border health care in the European union: challenges and opportunities. PMID- 17405245 TI - Cow 54, where are you? Producer liability and the national animal identification system. PMID- 17405246 TI - The 2004 Organ Donation Recovery and Improvement Act: how Congress missed an opportunity to say "yes" to financial incentives for organ donation. PMID- 17405247 TI - A chronic concern no more: how federal medical malpractice caps will survive under the Equal Protection Clause of the United States Constitution. PMID- 17405248 TI - Meeting the challenges of neurosurgery. The 2006 presidential address. AB - The challenges faced by neurosurgery in 2006 are many. There are five principal challenges that have received a great deal of attention from the Board of Directors of the American Association of Neurological Surgeons this past year. These are the challenge of maintaining a modern, efficient, and responsive educational program for neurosurgery; the challenge of maintaining the boundaries of neurosurgical practice and preventing the incursion of subspecialty groups into the performance of neurosurgical procedures; the challenge of responding to the changing demands of society; the challenge of influencing increased reimbursement; and the challenge of creating meaningful medical liability reform. Each of these issues are discussed. PMID- 17405249 TI - Health economics and the search for shunt-responsive symptomatic hydrocephalus in the elderly. PMID- 17405250 TI - Shunts in normal-pressure hydrocephalus: do we place too many or too few? AB - OBJECT: The average 65-year-old patient with moderate dementia can look forward to only 1.4 quality-adjusted life years (QALYs), that is, longevity times quality of life. Some of these patients suffer from normal-pressure hydrocephalus (NPH) and respond dramatically to shunt insertion. Currently, however, NPH cannot be diagnosed with certainty. The authors constructed a Markov decision analysis model to predict the outcome in patients with NPH treated with and without shunts. METHODS: Transition probabilities and health utilities were obtained from a review of the literature. A sensitivity analysis and Monte Carlo simulation were applied to test outcomes over a wide range of parameters. Using shunt response and complication rates from the literature, the average patient receiving a shunt would gain an additional 1.7 QALYs as a result of automatic shunt insertion. Even if 50% of patients receiving a shunt have complications, the shunt response rate would need to be less than 5% for empirical shunt insertion to do more harm than good. Authors of most studies have reported far better statistics. CONCLUSIONS: In summary, many more patients with suspected NPH should be considered for shunt insertion. PMID- 17405251 TI - Screening tests for normal-pressure hydrocephalus: sensitivity, specificity, and cost. AB - OBJECT: Many tests have been proposed to help choose candidates for shunt insertion in cases of suspected normal-pressure hydrocephalus (NPH). It is unclear what sensitivity and specificity a prospective test must have to improve outcomes, compared with the results of automatic shunt insertion. METHODS: The authors adapted the decision analysis model used in a companion article to allow for application of a screening test. Using the reported sensitivities and specificities of several such tests, they evaluated the effects such tests would have on the expected outcome of an average 65-year-old patient with moderate dementia. They also evaluated the cost-effectiveness of a theoretical screening test with superior sensitivity and specificity. CONCLUSIONS: Although external lumbar drainage comes quite close, none of the screening tests reported to date have sufficient sensitivity and specificity to improve expected outcome in an average candidate, compared with the results of automatic shunt placement in cases of suspected NPH. In addition, even a theoretically improved test would need to be considerably less expensive than prolonged lumbar drainage to be cost effective in clinical practice. PMID- 17405252 TI - Physiological and clinical advantages of median nerve fascicle transfer to the musculocutaneous nerve following brachial plexus root avulsion injury. AB - OBJECT: Loss of biceps muscle function is a significant disability after brachial plexus root avulsion injuries. Nerve grafting techniques to reestablish anatomical and functional continuity between the spinal cord and the avulsed root have not proven successful. Using nerve transfers for functional restoration of root avulsion injuries appears to be effective and has physiological advantages for reducing regeneration distances. Since the early 1990s, the Oberlin technique of transferring ulnar nerve fascicles to the motor branch of the musculocutaneous nerve has been the preferred operative technique for reinnervation and restoration of biceps muscle function. In the current study the authors examine the efficacy of an alternative technique using median nerve fascicles transferred to the musculocutaneous nerve to reinnervate the biceps muscle. METHODS: Forty consecutive patients with combined C5-6 brachial plexus root avulsions were evaluated pre- and postoperatively according to the British Medical Research Council Motor Grading Scale. Personal interviews concerning quality of life (QOL) after surgery were conducted and scored based on standards set by the World Health Organization. All patients showed some degree of improvement in biceps muscle function. Thirty-six (90%) of the 40 patients regained movement against gravity. The patients had a 77% improvement in overall QOL after the surgery; most notably, 92% of the patients reported their lack of need for medication and 75% a significant lessening of postoperative pain. Redirection of part of the healthy median nerve resulted in no measurable functional deficits, and only 28 patients reported minor sensory disturbances in the first web space for an average of 3 months after surgery. CONCLUSIONS: Median nerve fascicle transfer resulted in a significant improvement in biceps muscle function with an acceptable level of morbidity and should be considered an effective, and in many cases preferable, alternative to ulnar nerve fascicle transfer. PMID- 17405253 TI - Use of clinical signs and computed tomography myelography findings in detecting and excluding nerve root avulsion in complete brachial plexus palsy. AB - OBJECT: The purpose of this study was to investigate the usefulness of preoperative evaluation based on clinical testing and computed tomography (CT) myelography in differentiating root rupture (that is, graftable root) from root avulsion in total brachial plexus palsy. METHODS: Thirty-two patients with total brachial plexus palsy were clinically tested for the presence of phrenic nerve palsy, supraclavicular Tinel sign, shoulder protraction, Bernard-Horner syndrome, and hand pain. The patients underwent CT myelography and then underwent surgery. The combination of a positive Tinel sign and a positive shoulder protraction test accurately predicted the presence of a graftable root in 93.7% of the cases. A 96.8% rate of accuracy was attained if the results of the CT myelography were considered together with the clinical signs. The presence of Bernard-Horner syndrome and hand pain accurately indicated avulsion of the lower roots in 93.7% of the patients. Computed tomography myelography accurately predicted the condition of the lower roots in 100% of the cases. Total avulsion injury was observed in five cases (16%). The lower roots were avulsed in 94% of the cases. The C-5 and C-6 roots were grafted 40 times, and a suitable root stump for grafting lay in a retroscalenic position in 18 (45%) of the 40 cases. CONCLUSIONS: Preoperative assessment based on clinical examination and CT myelography allowed correct surgical planning in more than 90% of the cases. PMID- 17405254 TI - Systemic metabolic effects of combined insulin-like growth factor-I and growth hormone therapy in patients who have sustained acute traumatic brain injury. AB - OBJECT: Hypermetabolism, hypercatabolism, refractory nitrogen wasting, hyperglycemia, and immunosuppression accompany traumatic brain injury (TBI). Pituitary dysfunction occurs, affecting growth hormone (GH) and plasma insulin like growth factor-I (IGF-I) concentrations. The authors evaluated whether combination IGF-I/GH therapy improved metabolic and nutritional parameters after moderate to severe TBI. METHODS: The authors conducted a prospective, randomized, double-blind study comparing combination IGF-I/GH therapy and a placebo treatment. Ninety-seven patients with TBI were enrolled in the study within 72 hours of injury and were assigned to receive either combination IGF-I/GH therapy or placebo. All patients received concomitant nutritional support. Insulin-like growth factor-I was administered by continuous intravenous infusion (0.01 mg/kg/hr), and GH (0.05 mg/kg/day) was administered subcutaneously. Placebo control group patients received normal saline solution in place of both agents. Nutritional and metabolic monitoring continued throughout the 14-day treatment period. The two groups did not differ in energy expenditure, nutrient intake, or use of insulin treatment. The mean daily serum glucose concentration was higher in the treatment group (123 +/- 24 mg/dl) than in the control group (104 +/- 11 mg/dl) (p < 0.03). A positive nitrogen balance was achieved within the first 24 hours in the treatment group and remained positive in that group throughout the treatment period (p < 0.05). This pattern was not observed in the control group. Plasma IGF-I concentrations were above 350 ng/ml in the treatment group throughout the study period. Overall, the mean plasma IGF-I concentrations were 1003 +/- 480.6 ng/ml in the treatment group and 192 +/- 46.2 ng/ml in the control group (p < 0.01). CONCLUSIONS: The combination of IGF-I and GH produced sustained improvement in metabolic and nutritional endpoints after moderate to severe acute TBI. PMID- 17405255 TI - Effectiveness and handling characteristics of a three-layer polymer dura substitute: a prospective multicenter clinical study. AB - OBJECT: The goal of this study was to assess the effectiveness and handling characteristics of a dura substitute composed of two outer layers of expanded polytetrafluoroethylene (PTFE) and a middle layer consisting of an elastomeric fluoropolymer. METHODS: In a prospective multicenter study, the dura substitute was implanted using a standard technique in 119 patients undergoing cranial or spinal surgery requiring duraplasty. Intraoperative assessments of the dura patch consisted of testing for cerebrospinal fluid (CSF) leakage employing the Valsalva maneuver and a surgeon's standard evaluation of the handling characteristics of the device. Postoperative assessments conducted during a mean follow-up time of 15.7 months (range 0.3-45.6 months) consisted of physical examinations, routine computed tomography (CT) or magnetic resonance (MR) imaging studies, and histological studies of any removed dura patches. The mean age of the 119 patients was 40 years (range < 1-81 years). The dura substitute was implanted cranially in 102 patients and spinally in 17. Intraoperative assessment including the Valsalva maneuver led to application of additional sutures in 17 patients. Handling features were rated very good to excellent. Postoperative clinical evaluation resulted in 79 excellent and 18 good results. Imaging studies (MR imaging studies in 69 patients and CT studies in 34 patients) showed no adhesions in 87 patients and minimal adhesions in seven patients (the dura was not visualized in nine patients). Postoperative complications occurred in 12 patients. There were six cases of CSF leakage, three cases of extradural hematoma, one case of arachnoid fibrosis after decompression of a Chiari malformation Type I, and two cases of infection. Eight (7%) of these complications were potentially related to the dura patch. CONCLUSIONS: In a large, multicenter clinical study of the use of an expanded-PTFE-containing dura substitute, the device was found to be easy to handle and implant. No serious dura patch-related intraoperative adverse events were observed. Postoperatively, there were no major sealing problems or long-term complications. In two cases the patch had to be removed due to fibrosis and infection. The three-layer polymer dura substitute appears to be safe and effective in minimizing CSF leakage and adhesion formation, and its use avoids any risk of prion disease transmission. PMID- 17405256 TI - The role of recombinant activated factor VII in neurosurgery: hope or hype? AB - Recombinant activated factor VII (rFVIIa) is a relatively new pharmaceutical agent developed for use in patients with hemophilia in whom inhibitors to clotting factors VIII or IX have developed. Use of this drug has become common in recent years because of its efficacy and safety in patients with coagulation disorders as well as in patients who are at high risk for thromboembolism, even when other means of establishing hemostasis have failed. The use of rFVIIa in neurosurgery has lagged behind its use in other fields, although there is a growing body of literature on such uses. In this article the authors review the history and science of rFVIIa as well as dosing and safety information. Various uses pertinent to the neurosurgeon are reviewed, including the treatment of patients with coagulation disorders, those suffering trauma, and those with perioperative hemorrhage, intracerebral hemorrhage, or subarachnoid hemorrhage. Based on their review of the uses of rFVIIa, the authors conclude that rFVIIa is a safe and effective agent with the potential to revolutionize the treatment of neurosurgical patients with hemorrhage. Cost is a major impediment to the widespread use of rFVIIa, and there is some evidence that its use in the neurosurgical population may be subject to higher risk than in other populations studied thus far. Although further study is needed to better delineate the safety and efficacy of the drug in many nonlicensed uses, it is clear that rFVIIa is an agent with tremendous promise. PMID- 17405257 TI - Minimally invasive implantation of the peritoneal catheter in ventriculoperitoneal shunt placement for hydrocephalus: analysis of data in 151 consecutive adult patients. AB - OBJECT: The authors report on their experience with laparoscopy-guided implantation of a peritoneal catheter in ventriculoperitoneal shunt placement procedures in adults. METHODS: In performing the conventional method of shunt placement in 2001, 8% of the cases resulted in malposition and dislocation of the distal catheter; therefore, the authors together with personnel from the Department of General Surgery decided to utilize an interdisciplinary approach involving laparoscopy-guided implantation of the catheter. Between October 2001 and January 2005, 202 ventriculoperitoneal shunt placement procedures were conducted in adult patients for hydrocephalus of various origins. In 152 patients, laparoscopy-guided implantation of the distal catheter was performed. In all except one of these patients, implantation was successful. Laparoscopy and the cranial part of the surgery were performed simultaneously. There was an 8% rate of malposition of the distal catheter in the nonlaparoscopy group. In contrast, there was no dislocation or malposition of the distal catheter in the laparoscopy group. Two cases (1.3%) of shunt infection occurred in the laparoscopy group. CONCLUSIONS: Laparoscopic implantation of a distal catheter is a simple, minimally invasive, and easy procedure to perform and allows exact localization of the peritoneal catheter and confirmation of its patency. PMID- 17405258 TI - Resection of benign sciatic notch dumbbell-shaped tumors. AB - OBJECT: The operative management of combined intrapelvic and extrapelvic sciatic notch dumbbell-shaped tumors is challenging. The relatively rare occurrence of these tumors and the varied extent of disease have made it difficult for surgeons to establish definitive surgical indications or predict favorable neurological outcomes based on preoperative imaging data. METHODS: In the past 3 years, the authors treated five patients presenting with radiating leg pain as a result of benign sciatic notch dumbbell-shaped tumors. These tumors in three patients with unilateral leg symptoms were considered unresectable by other neurosurgeons because of presumed direct intrinsic neural involvement. After high-resolution magnetic resonance (MR) imaging demonstrated that the extensive tumors were separate from the sciatic nerve and the lumbosacral plexus, however, these patients underwent a combined one-stage transabdominal and posterior transgluteal complete resection. Normal neurological status was maintained postoperatively in these three patients, and after more than 1 year of postoperative follow up, there were no tumor recurrences. In two patients with bilateral symptoms and extensive tumor burden, serial MR images showed that innumerable tumors directly involved the entire cross-sectional area of the sciatic nerves and extended longitudinally to the lumbosacral plexuses. Tumor debulking or resection in these patients would have resulted in neurological deficits and would not have addressed their neuropathic pain, and therefore no surgery was performed. These two patients were treated pharmacologically and advised to monitor their tumor status over the course of their lifetimes in case of malignant transformation of the tumor. CONCLUSIONS: A combined one-stage transabdominal and transgluteal approach allows safe resection of selected benign but extensive sciatic notch tumors. High-resolution MR imaging is a useful tool in the management of these tumors because it allows the surgeon to visualize the anatomical relationships of the tumor to the sciatic nerve. The authors believe that as this imaging technology advances, it will provide surgeons with a method to predict definitively which sciatic notch tumors displace rather than directly involve the sciatic nerve, and therefore indicate which tumors can be resected safely and completely. PMID- 17405259 TI - Quantitation of the lower subscapular nerve for potential use in neurotization procedures. AB - OBJECT: New information regarding nerve branches of the brachial plexus can be useful to the surgeon performing neurotization procedures following patient injury. Nerves in the vicinity of the axillae have been commonly used for neural grafting procedures, with the exception of the lower subscapular nerve (LSN). METHODS: The authors dissected and measured the LSN in 47 upper extremities (left and right sides) obtained in 27 adult cadavers, and determined distances between the LSN and surrounding nerves to help quantify it for possible use in neurotization procedures. The mean diameter of the LSN was 2.3 mm. The mean length of the LSN from its origin at the posterior cord until it branched to the subscapularis muscle was 3.5 cm, and the mean distance from this branch until its termination in the teres major muscle was 6 cm. Therefore, the mean length of the entire LSN from the posterior cord to the teres major was 9.5 cm. When the LSN was mobilized to explore its possible use in neurotization, it reached the entrance site of the musculocutaneous nerve into the coracobrachialis muscle in all but three sides and was within 1.5 cm from this point in these three. In the other specimens, the mean length of the LSN distal to this site of the musculocutaneous nerve was 2 cm. The mobilized LSN reached the axillary nerve trunk as it entered the quadrangular space in all specimens. The mean length of the LSN distal to this point on the axillary nerve was 2.5 cm. Furthermore, on all but one side the LSN was found within the confines of an anatomical triangle previously described by the authors. CONCLUSIONS: The authors hope that these data will prove useful to the surgeon for both identifying the LSN and planning for potential neurotization procedures of the brachial plexus. PMID- 17405260 TI - Effects of ethanol on limited resuscitation in a model of traumatic brain injury and hemorrhagic shock. AB - OBJECT: Limited resuscitation following uncontrolled hemorrhagic shock (HS) has been associated with improved outcomes in various animal models, although it has not been previously studied in the setting of traumatic brain injury (TBI) and ethanol intoxication. The aim of the present study was to determine the effects of ethanol intoxication in a model of experimental TBI and HS treated with limited resuscitation. METHODS: After induction of anesthesia and the placement of instruments, swine were subjected to a fluid-percussion injury of 3 atm. Simultaneously, hemorrhage was induced from an arterial catheter via a computerized roller pump to a mean arterial blood pressure (MABP) of 50 mm Hg, at which time uncontrolled hemorrhage was induced by the creation of an aortic tear. When the MABP decreased to 30 mm Hg, limited resuscitation to a MABP of 60 mm Hg was begun. After 60 minutes, animals were aggressively resuscitated to baseline MABP levels. Two groups of animals were studied: those receiving tap water by gastrostomy tube and those receiving ethanol (4 g/kg) by gastrostomy tube. Animals were monitored for 180 minutes after TBI. Hemorrhage volumes were significantly greater in ethanol-infused animals (mean +/- standard deviation, 41 +/- 34 mm Hg) compared with tap water-infused animals (17 +/- 18 mm Hg; p = 0.048). Resuscitation requirements were significantly higher and metabolic parameters significantly worse in the ethanol group. Survival time was also significantly decreased in the animals infused with ethanol (81 +/- 60 minutes) compared with those infused with tap water (130 +/- 51 minutes; p = 0.035). CONCLUSIONS: Ethanol intoxication led to increased hemorrhage volume and worsened hemodynamic and metabolic profiles in this model of limited resuscitation after TBI and HS. Ethanol-exposed animals had increased resuscitation requirements and decreased survival times. PMID- 17405261 TI - Long-term electrical stimulation-induced inhibition of partial epilepsy. Case report. AB - The aim of this study was to determine the effects of long-term continuous cerebrocortical electrical stimulation in the treatment of partial epilepsy. The authors review the case of a 44-year-old man with medically intractable postencephalitic localization-related epilepsy with ictal onset in the primary motor cortex. For 5 years he was treated using patterned subthreshold electrical stimulation of the ictal site. This therapy has successfully eliminated the jacksonian march of cortical excitability and secondary generalization and reduced seizure frequency and intensity with an immediate postictal return of motor function. Over time, the seizure frequency subsided by more than 90%, with the patient showing no adverse features resulting from focal stimulation. The results in this case support the hypothesis that effective and safe long-term modulation of focal epilepsy is possible with focal cerebrocortical electrical stimulation. PMID- 17405262 TI - Boron neutron capture therapy for recurrent malignant meningioma. Case report. AB - Malignant meningioma is a rare brain tumor with a high risk of recurrence. If this tumor recurs after complete resection and adjuvant radiotherapy, there is no optimal treatment to control it. The authors report the first case of recurrent malignant meningioma treated using boron neutron capture therapy (BNCT). This 25 year-old pregnant woman presented with a large frontal tumor. After her baby was born, she underwent gross-total resection of the tumor. A second resection and three Gamma Knife surgeries could not control progression of the enhancing mass; therefore, the authors applied BNCT based on their experience with it in the treatment of malignant gliomas. The minimum tumor dose and maximum brain tissue dose were estimated as 39.7 Gy-Eq and less than 9.0 Gy-Eq, respectively. Before BNCT the patient was mobile by wheelchair only, whereas 1 week after therapy she was able to walk. Twenty-two weeks later she underwent a second BNCT for tumor regrowth on the contralateral side, and the lesion was subsequently reduced. The tumor volume was markedly decreased from 65.6 cm3 at the time of the first BNCT to 31.8 cm3 at 26 weeks thereafter. The treatment of recurrent malignant meningioma is difficult and has been discouraging thus far. Data in the present case indicate that BNCT may be a promising treatment option for this challenging tumor. PMID- 17405263 TI - Gliosarcoma with infratemporal fossa extension. Case report. AB - Gliosarcomas are bimorphic intraaxial tumors. Involvement of the skull base is highly unexpected. The authors present the case of a temporal lobe gliosarcoma with significant infratemporal fossa extension. This 55-year-old man presented with a 1-month history of severe progressive headache. Neurological examination was unremarkable except for bilateral papilledema. Magnetic resonance imaging revealed a 6-cm right temporal mass with extension into the infratemporal fossa. The patient underwent a right frontotemporal craniotomy together with drilling of the sphenoid ridge and middle fossa floor. The tumor consisted of intraaxial, intracranial as well as extradural, and extracranial components with extension to the posterolateral wall of the sphenoid sinus. It had a relatively well circumscribed dissection plane. Gross-total resection was achieved, and the middle fossa floor was reconstructed using a rotated temporalis muscle flap. The postoperative course was uneventful except for hypesthesia in the distribution of the maxillary division of the right trigeminal nerve. The histopathological diagnosis was consistent with gliosarcoma. Radiotherapy and chemotherapy consisting of temozolomide were administered subsequently, and the patient was recurrence free 12 months after his initial diagnosis. In the presence of a mass lesion with both intraaxial and extracranial involvement, gliosarcoma should be considered among the differential diagnoses. Aggressive resection should be attempted, including the use of skull base surgical techniques to ensure an optimal outcome. The effect of skull base involvement to the overall treatment and outcome of patients with gliosarcomas would be difficult to determine given the rare occurrence of these lesions in such locations. PMID- 17405264 TI - Rare giant cell ependymoma in an octogenarian. Case report and review of the literature. AB - Ependymomas are glial tumors that occur most often in children. In adults, ependymomas most often appear in the spinal cord. The World Health Organization recognizes several rare ependymoma subtypes, including the giant cell ependymoma of the terminal filum. The authors describe an unusual case of a posterior fossa giant cell ependymoma in an 89-year-old man presenting with vertigo and disequilibrium. Only seven cases of this tumor have been reported in the literature to date. The authors discuss the clinical presentation, radiological findings, pathological considerations, and surgical intervention in this patient and review the relevant literature. PMID- 17405265 TI - Granulocytic sarcoma: an unusual complication of acute promyelocytic leukemia causing cerebellar hemorrhage. Case report. AB - Granulocytic sarcomas are rare tumors that occur primarily in patients with acute myelogenous leukemia or other myeloproliferative disorders, are seldom seen in patients with acute promyelocytic leukemia (APL), and have never been reported to occur in the cerebellum. The authors describe the case of a patient with APL who harbored a hemorrhagic granulocytic sarcoma in the cerebellum. This 39-year-old woman presented with cerebellar ataxia. Magnetic resonance images revealed an intraaxial tumor in the cerebellum. Bone marrow samples showing infiltration by leukemic blast cells and data from hematological tests led to a diagnosis of APL. The patient was treated with chemotherapy and surgery. She had no response to chemotherapy and died of progressive intratumoral hemorrhage. Results of histopathological studies and immunohistochemical staining of the cerebellar tumor confirmed a granulocytic sarcoma. Flow cytometry showed that the blast cells were positive for leukocyte common antigen, CD13, and CD33 markers. Bone marrow cytogenetics revealed that the patient had a 46,XX karyotype. Although no cytogenetic abnormality was present, fluorescence in situ hybridization detected a chimeric fusion of PML and RARA. This is the first report to document a granulocytic sarcoma in the cerebellum as the primary presentation in a patient with APL and abnormal coagulation. As predicted by the unusual clinical manifestations and radiological findings, the patient's survival was short. Although central nervous system complications in patients with APL are rare, the data in this case highlight the need for individualized treatment when such conditions occur. PMID- 17405266 TI - Myeloid sarcoma with multiple lesions of the central nervous system in a patient without leukemia. Case report. AB - The authors report the unusual case of a 35-year-old woman suffering from left leg numbness and radiculopathy due to multiple lesions in the central nervous system: one right parietal extracranial-intracranial lesion with invasion of the sensory cortex, and two intraspinal, intradural lesions compressing the spinal cord at T3-5 and S 1-4. Biopsy sampling of the extracranial part of the parietal lesion led to a diagnosis of myeloid sarcoma. Further examination revealed no evidence of leukemic disease or myeloproliferative disorder. An aggressive multimodal approach to treatment in this patient with a combination of chemotherapy, whole-body radiotherapy, and allogeneic bone marrow transplantation was started immediately. The patient experienced full neurological recovery and complete disappearance of all lesions. At the 7-year follow-up examination, there was no evidence of disease. To the authors' knowledge, this is the first report of a myeloid sarcoma with both intracranial and intraspinal manifestations in a patient without leukemia. PMID- 17405267 TI - Using an end-to-side interposed sural nerve graft for facial nerve reinforcement after vestibular schwannoma resection. Technical note. AB - Increasing rates of facial and cochlear nerve preservation after vestibular schwannoma surgery have been achieved in the last 30 years. However, the management of a partially or completely damaged facial nerve remains an important issue. In such a case, several immediate or delayed repair techniques have been used. On the basis of recent studies of successful end-to-side neurorrhaphy, the authors applied this technique in a patient with an anatomically preserved but partially injured facial nerve during vestibular schwannoma surgery. The authors interposed a sural nerve graft to reinforce the facial nerve whose partial anatomical continuity had been preserved. On follow-up examinations 18 months after surgery, satisfactory cosmetic results for facial nerve function were observed. The end-to-side interposed nerve graft appears to be a reasonable alternative in cases of partial facial nerve injury, and might be a future therapeutic option for other cranial nerve injuries. PMID- 17405268 TI - Endoscopic opening of the foramen of magendie using transaqueductal navigation for membrane obstruction of the fourth ventricle outlets. Technical note. AB - A membrane obstruction of the foramina of Magendie and Luschka is an uncommon origin of hydrocephalus characterized by unusual clinical symptoms of rhomboid fossa hypertension. Various surgical approaches have been proposed to alleviate this obstruction, including opening the obstructed foramen of Magendie using suboccipital craniectomy, shunting procedures, and more recently, endoscopic third ventriculostomy (ETV). In some cases, however, reshaping of the posterior fossa due to the collapse of the prepontine cistern could make ETV difficult for the surgeon and dangerous to the patient. In these cases, endoscopic opening of the foramen of Magendie by transaqueductal navigation of the fourth ventricle is a suitable and feasible therapeutic option. PMID- 17405269 TI - The use of waterjet dissection in endoscopic neurosurgery. Technical note. AB - Waterjet dissection enables vessel preservation and a reduction in intraoperative blood loss. Because even minimal bleeding should be avoided during neuroendoscopy, the waterjet device may be a particularly valuable tool in such procedures. The authors used this instrument in experimental endoscopic procedures in 20 cadaveric porcine brains and clinically in four patients with obstructive hydrocephalus. A precise and accurate septostomy was achieved in all of the pig brains. In two patients the hydrocephalus was due to intraventricular hemorrhage, in one a posterior fossa tumor, and in one a cystic craniopharyngioma. In all patients the surgical view was kept clear with waterjet irrigation and suction. Using a pressure setting of 10 bars, the waterjet device successfully perforated the cyst wall of the craniopharyngioma in one patient and the floor of the third ventricle in three patients. The use of the waterjet device in selected endoscopic procedures appears safe, and may help reduce intraoperative bleeding. However, further studies are needed to confirm the utility of the waterjet tool in endoscopy. PMID- 17405270 TI - Abnormal enhancement on imaging studies preceding hyperperfusion syndrome. Case illustration. PMID- 17405271 TI - Solitary choroid plexus tuberculoma in an adult patient. Case illustration. PMID- 17405272 TI - Spontaneous pneumocephalus caused by the association of pneumosinus dilatans and meningioma. Case illustration. PMID- 17405273 TI - [Argon--a biologically active component of atmosphere]. AB - Analysis of the literary and author's own data denies physiological effects of argon in normobaric normoxic atmosphere that could impact human performance, and embryogenesis of lower vertebrates. In hypoxic atmosphere, argon keeps up the normal oxygen consumption by human and rodent organisms, augments the working ability of humans, and exacerbates the hypoxia-driven depression of lower vertebrates embryogenesis. Potential anti-hypoxia benefits from argon introduction into artificial atmosphere are considered. PMID- 17405274 TI - [Viscously-elastic properties of brain and loss of consciousness by human under acceleration]. AB - The article presents the results of analysis of available empirical data about human tolerance of high g-loads. Consideration is given to the role of cerebral viscous elasticity and brain and skull stem deformations in loss of consciousness due to long-term exposure to +Gz and +Gx loads including g-loads with a high rate of onset. High g-loads and rates of onset can lead to maximum permissible g-dose (delta V)m = g integral(n - 1)dt [M/c] = const invariant with a maximum g-dose or rate of onset. For the head-pelvis and chest-back g-vectors it amounts to (deltaVz)max approximately equal 400 M/c; (nz = 2...15; nz = 2...7,5 g/c) and (deltaVx)max =(10...12)10(3) M/c; (nx = 12..22; nx< or = 0,5g/c), respectively. It is hypothesized that maximum permissible gdose for humans are constants which, probably, characterize buildup of brain damaging() stresses and deformations and ensuing loss of consciousness as a reaction of the brain, stem structures spedfically, to the g-load induced drop of arterial blood and intracranial pressure, and also deterioration of cerebral viscous elasticity (turgor). PMID- 17405275 TI - [Effect of long-term microgravity on the vestibular function]. AB - Comprehensive computerized oculomotor testing was used to investigate the vestibular function in 9 Russian members of ISS crews 3-9 on days 1 (2), 4 (5) and 8 (9) of return from long-term stay in microgravity (126 to 195 days). The vestibular function was assessed by the static otolith-cervical-ocular reflex, dynamic otolith-cervical-ocular reactions, vestibular reactivity, and spontaneous oculomotor activity. The postflight investigations revealed functional disorders in the peripheral (an increased vestibular reactivity, absent or damped otolith cervical-ocular reflex), and central (spontaneous typical and atypical nystagmus, gaze nystagmus) vestibular analyzer. The pattern and extent of vestibular disorders after long-term exposure in microgravity were individual by character; however, some of the vestibular reactions, including disappearance or considerable damping of the static otolith-cervical-ocular reflex, exaggerated vestibular reactivity and spontaneous eye movements, displayed consistency. PMID- 17405276 TI - [Blood plasma proteins following long-duration space flight]. AB - Protein composition of blood plasma was an object of investigation in 29 Russian cosmonauts flown on the MIR station from 125 to 366 days. Protein fractions were analyzed using acetate cellulose electrophoresis. Concentration of total protein was determined with the help of the biuret reaction on an automated analyzer. On the second day post flight, mean concentration of total protein and percentage of the protein fractions were equal to baseline values. In the interval between days 7 and 14 post flight, total protein was statistically reduced, alphal- N alpha2 globulins increased and y-globulin reduced, whereas albumin and beta-globulins were unchanged in the average. These results may point to development of an acute reaction in the early period of readaptation to the return from long-duration space flight. PMID- 17405277 TI - [Cutaneous electro-gastroenterography for evaluation of changes in the gastrointestinal tract in consequence of various angles of inclination and length of head-down tilt]. AB - Experimental results were compared with data of cutaneous electro gastroenterography and USI of the gastrointestinal tract in human subjects at different tilt angles and after different periods of tilting. It was shown that increased GIT electrical activity in tilted subjects before meal is representative of increasing intracavetary contents of the stomach and small intestine. More intensive electrical activity results from elevation of the GIT tone and peristalsis. Extension of tilt duration raises GIT tone and peristalsis and, consequently, stimulates the GIT electrical activity. Further increase of the tilt angle suppresses evacuation and, therefore, the electrical activity Analysis of the results allows conclusion that increased parameters of the total energy of GIT electrical potentials are reflective of elevated tone of GIT walls, whereas elongation of the potential peaks envelope implies activation of peristalsis. PMID- 17405278 TI - [The combined action of octafluoropropane and bipolar ionized air]. AB - An experiment with Wistar male rats was to look into the action of octafluoropropane (OFP, of 50 microg/m(3)) combined with bipolar ionized air (BIA) at a concentration of light air ions of 60,000 para-air in cm(3). The chamber experiment was 43 days long. Intoxication by OFP reduced body mass, as well as the erythrocyte count and hemoglobin level in peripheral blood. As for BIA, it appeared to bring these indices back to their normal values. Erythrocyte metabolism underwent phase-by-phase shifts; but breathing BIA mitigated these shifts markedly. As regards to erythrocyte metabolism in intoxicated animals, BIA had a compensatory effect Changes in the biochemical profile of blood plasma make us think, that BIA counteracts the OFP damaging action on the myocardium and, at the same time, aggravates impairment of metabolism in the liver and, probably, kidney. Variations in the spectrum and total content of higher fatty acids in the lung in the experiment were more pronounced in the event of exposure to OFP+BIA than to OFP alone. Also, the combined exposure increased the level of laurinic acid. Histological investigations of the liver, spleen, myocardium, trachea and the lung attested to the dystrophic damage of the liver, spleen plethora and reticular hyperplasia, and slight cloudy swelling of the myocardium attributed to OFP. After 14 days since the end of the experiment, histological changes were much less dramatic; in 39 days after the experiment all the changes were gone with the exception of weak emphysematosis. BIA had no effect on animals in the absence of OFP; neither was there any significant difference between control (intact) animals and those who breathed BIA in laboratory. To conclude, 50 microg/m(3) of OFP which falls far short of the existing maximum permissible levels, a strong toxic action on animals. Aside from mitigation of the OFP toxic action on erythrocytes and myocardium, BIA, when breathed with a long time, aggravated the metabolic disorders in the liver and lung provoked by OFP. PMID- 17405279 TI - [Role of human olfaction in detection of space vehicle air contamination by gases emitted during thermal destruction of polymers]. AB - The paper deals with the ability of human olfaction to identify odours associated with thermal destruction of polymers. The investigations confirmed the ability of early detection of equipment overheating and subjective odour evaluation using the 5-point intensity scale and calculating the relation of odour intensity to the maximum admissible concentration of volatile chemical contaminants aboard piloted space vehicles. Emission of gases by polymers in electrical equipment starts with heating temperature rise to 100 degrees C. A new odour smelled by only some of humans due to individual perception signifies typically that emitted gases are below MAC for space vehicle. Odour sensed by everyone though not irritating signifies, as a rule, of chemical contamination above MAC but not contingency. Symptoms of irritation by products of thermal destruction of polymers signify contingency and require the use of personal protection equipment. PMID- 17405280 TI - [Some aspects of water electrolysis with the use of a solid polymer electrolyte]. AB - Electrochemical process in cells with a solid polymer electrolyte is dependent on catalyst durability in harsh environments and catalyst sputtering technology to ensure efficient power consumption. Active polymer electrolytes will permit to reduce substantially non-productive layouts and design a cost-effective, compact and safe system generator of high-purity oxygen and hydrogen. The existing designs of combined oxide systems integrating rear-earth and earth metals with a structure of Ln3+x Me2+1-x CoO3 containing perofskites were shown to be active catalysts in cells with a solid polymer electrolyte, and the sputtering technology was proven to reduce non-productive layouts in 2 or 2.5 times. PMID- 17405281 TI - [Determination of the intelligence quotient of pilots with incipient atherosclerosis]. AB - Comprehensive examination, including clinical-functional and psychological testing, was given to 189 essentially healthy civil pilots and 235 pilots with atherosclerosis of aorta and trunks without considerable blood flow disturbance. The total of 835 investigations was performed. Distribution into health groups was conducted on clinical diagnosis. Pilots with cardiovascular pathologies were found to have the intelligence quotient significantly lowered. Associated clinical and psychological tests were effective in revealing and dynamic monitoring of incipient diseases, and taking reasoned disposition regarding pilot's fitness for flight duties. PMID- 17405282 TI - [System-psychological evaluation of cosmonaut's efficiency]. AB - In view of criticality of objective diagnostics of mental performance and thereupon prediction of operational efficiency of crew members for safety and success of space mission, Institute for Biomedical Problems undertook a series of investigations towards the development of a system-psychological concept as a basis for practical diagnostics of mental performance of crew on a mission. The investigations made it possible to foresee the work style and probability of successful fulfillment of manual vehidce control using the knowledge of the types of mental performance. PMID- 17405283 TI - [Time of euglobulin coagulation as a short-cut method of blood aggregation diagnostics]. AB - Euglobulin coagulation time (ECT) forms the basis for a short-cut method of assessing the blood activator potential, a pivot of fibrinolysis which is responsible for blood fluidity. During simulation of complicated flight conditions the method was good enough to display variations in the blood activator potential in response to specific types of extreme factors. ECT was measured in 94.47% of 398 patients subjected to hemocoagulation examination. In 338 patients ECT was within the clinical norm and in 60 patients ECT was in the pathology range. Complex analysis of experimental and clinical coagulograms including ECT, fibrinolysis, factor I, prothrombin time, and prothrombin index brought to the conclusion about cardiovascular and hepatic dysfunctions. Therefore, our investigations showed that the ECT short-cut method can be adopted for blood aggregation diagnostics in military and civil pilots. PMID- 17405284 TI - [Human science in aviation of the XXIst century]. PMID- 17405285 TI - [Egineering psychology and ergonomics in aviation]. PMID- 17405286 TI - [Bone grafts (editorial)]. AB - Bone grafts remains the most effective grafting material because it provides the three elements required for bone regeneration: osteoconduction, osteoinduction and osteogenic cells. Bone grafts are used in the repair of significant fractures, the treatment of skeletal tumors and the reconstruction of failed total arthroplasties. Allografts of bone elicit transplantation immunity and this immunogenicity may cause the higher failure rate of these grafts. In an attempt to reduce the immunogenicity of the graft, allogenic bone has been frozen and freeze-dried. Because of increasing clinical use of bone grafts and the unacceptable failure rate, it would be advantageous to better understand the biological response to grafts in order to better define the parameters necessary for a successful bone grafts. PMID- 17405287 TI - [New multidisciplinary heart failure care program (six-month preliminary observation)]. AB - In the last years multidisciplinary care (MDC) of patients with chronic heart failure has proved to be effective. Because of specificity of the medical care organization in different countries the outcome of the programs may be various. The aim of this study was to evaluate the effect of MDC on patients' survival, hospitalization rate and stay. MATERIAL AND METHODS: 129 chronic heart failure patients admitted to Department of Cardiology were randomized to MDC or routine care (RC) groups. All patients were referred to their primary care physicians. In 64 patients from MDC group, aged 68 +/- 10.5 years, a follow-up was performed on 14 day, 3 and 6 months, on the basis of heart failure clinic, by cardiologist, heart failure nurse, psychologist and physiotherapist. Patients received heart failure nurse and cardiologist broad educational program systematically. Every day heart failure nurse telephone counseling and home-based interventions were also available to the patients. 65 patients from RC group, aged 72 +/- 11.5 years, were referred only to their primary care physicians. RESULTS: There was the lower duration of hospitalization in MDC than in RC group (8.3 +/- 5.31 vs 13.5 +/- 6.99 days; p < 0.05). A tendency to the lower duration of heart failure hospitalization and frequency of total and heart failure hospitalization in MDC group was observed (9.8 +/- 5.46 vs 12.1 +/- 6.23 days, 18.8 vs 25.0, and 12.5 vs 16.9% respectively). No significant differences were found between MDC and RC total and heart failure mortality (15.6 vs 16.9%, and 10.9 vs 12.3%, respectively). CONCLUSION: Six months multidisciplinary care of chronic heart failure patients has appeared more efficient than routine primary care. PMID- 17405288 TI - [Analysis of tryptase levels in infants and children with wheezy bronchitis]. AB - Wheezy bronchitis is one of hospitalization causes of infants and young children. Significant problem is recurrent wheezing. Wheezy bronchitis can be the first sign of bronchial asthma. New markers are taken into consideration in aspect of recurrent bronchitis prevention. Such marker seems to be tryptase. Tryptase is released from mastocytes in early phase of allergic reaction. The aim of the study was to assess tryptase and ECP concentrations among infants and young children with wheezy bronchitis. MATERIAL AND METHODS: Ninety-four patients with wheezy bronchitis in age from between 1 month to 36 months old were included into the study (47 with the first episode and 47 with at least third episode). Forty three patients hospitalized due to other causes, from the same age group (these patients haven't already had any wheezy bronchitis) were included into the control group. Among all patients concentrations of tryptase and ECP were evaluated (among patients from the study group in the acute phase of disease) by fluoroimmunoenzymatic method (FEIA) with the the use of UniCAP 100 set (Pharmacia & Upjohn Diagnostics AB). RESULTS: No statistically significant differences of tryptase concentrations in blood plasma during wheezy bronchitis among infants and young children in correlation to the control group were found. Statistically significant differences were identified in aspect of tryptase concentrations during wheezy bronchitis between patients with the first episode of wheezing and patients with recurrent wheezing and higher concentrations were observed among patients with the first episode. No statistically significant influence of family history of allergy and symptoms of allergy were identified in aspect of tryptase concentrations. CONCLUSIONS: Obtained results lead to conclusion that tryptase concentrations have a little significance in wheezy bronchitis. PMID- 17405289 TI - [Arginase activity in blood serum of patients with acute and chronic pancreatitis]. AB - Pancreatitis is the most common disease of the pancreas. Difficulties in early recognition of pancreatitic diseases, particularly chronic pancreatitis, are the reason of the search for new diagnostic methods. In our earlier studies we have shown that the determination of arginase activity in serum of patients with pancreatic cancer may be useful test in preoperative diagnosis of this cancer. The aim of this study was to asses the arginase activity in serum of patients with acute and chronic pancreatitis before and after medical treatment. MATERIAL AND METHODS: Arginase activity was studied in serum of 10 patients with acute and 10 patients with chronic pancreatitis obtained before, after and/or during the medical treatment. RESULTS: The increase of arginase activity was observed in both studied groups before the medical treatment, and a statistically significant decrease after the treatment (p < 0,05). There were not significant differences between arginase activity in acute and chronic pancreatitis. CONCLUSION: Arginase activity determination seems to be useful in monitoring the treatment of patients with acute and chronic pancreatitis. PMID- 17405290 TI - [Bile duct infections as a late complication after endoscopic sphincterotomy]. AB - Nowadays endoscopic sphincterotomy (SE) is considered to be a "gold standard" in the treatment of residual choledocholithiasis and biliary pancreatitis. Nevertheless patients after invasive procedures concerning bile ducts with destruction of Oddi's sphincter often have increased succeptibility to chronic infections of bile ducts. AIM: Assessment of the influence of SE on bacterial colonization, development of main bile duct chronic inflammation and recurrent choledocholithiasis. MATERIAL AND METHODS: We included to the study 50 patients with main bile duct lithiasis, who underwent endoscopic cholangiopancreatography (ERCP). Among them there were 32 women in age from 45 to 87 (mean 69.7 +/- 12.3) and 18 men in age from 39 to 84 (mean 59.8 +/- 11.56). The studied group was divided into 3 subgroups: In I (control) group were included 10 patients diagnosed with ERCP techniques because of jaundice of unknown origin. Those patients did't have ERCP or operative revision of bile ducts before, but all of them underwent cholecystectomy in the past. Each II and III group consisted of 20 patients, who underwent SE before 12-24 and 25-36 months earlier respectively. Sterile uodenoscope was inserted to the region of major duodenal papilla, and then a sterile brush was used to collect material to cytological smear (brushing) from main bile duct. In all groups 2 ml of bile were taken by a catheter to a syringe. On the basis of radiograms from ERCP findings, the width of main bile duct was assessed. In all patients blood cell count, aspartate and alanine aminotransferases, gammaglutamyltranspeptidase, alkaline phosphatase, amylase, bilirubin, C-reactive protein and procalcitonine levels were measured. RESULTS: In our study all 40 patients with previously performed endoscopic sphincterotomy (SE) had bacterial colonization in bile ducts, mostly with Gramm negative pathogens. The most virulent pathogens vere cultured from bile duct swabs from patients after two or more revisions of common bile duct, and after a longer time from sphincterotomy. Those patients had multibacterial bile duct infections (Escherichia coli + Pseudomonas aeruginosa; P. aeruginosa + Enterococcus faecalis; E. faecalis + Enterobacter cloacae + Candida albicans; P. aeruginosa + E. cloacae). E. coil and E. faecalis were most frequently cultured (20 and 16 cases respectively). It was observed that with length of time from endoscopic sphincterotomy, the width of main bile duct and also amount of cultured pathogens and their virulency grow. Elevated transaminases activity and serum procalcitonine levels were observed in studied groups. 9 patients after previous sphincterotomy had recurrent lithiasis. CONCLUSIONS: Sphincterotomy contributes to colonization of main bile duct with pathogenic bacteria. With length of time from endoscopic sphincterotomy, and amount of performed endoscopic sphincterotomies the amount of cultured pathogens, their virulency and drug resistance grow. Chronic infection of bile ducts after endoscopic sphincterotomy causes higher risk of recurrent lithiasis. PMID- 17405291 TI - [The analysis of lipidogram parameters and the parameters of coagulation and fibrynolysis systems as risk factors in overweight and obese children and adolescents]. AB - Since artherosclerosis processes were proved to have their origins as early as in one's childhood, attempts have been made to find the risk factors in the population of the developmental age. AIM: The aim of this work was the analysis of lipidogram parameters and the parameters of coagulation and fibrynolysis systems in overweight and obese children and adolescents. MATERIAL AND METHODS: 101 children and adolescents aged 4-19 were examined, including 30 overweight and 34 obese ones, with the control group comprising 37 patients with normal body weight. RESULTS: No significant differences in the studied groups were noticed in the total cholesterol level, LDL, fibrinogen and in alpha2-antiplasmin activity. The HDL cholesterol level was actually lower in the group with obese patients, while the PAI-1 level was actually higher. CONCLUSIONS: Overweight and obese children and adolescents comprise a group threatened by premature development of atherosclerosis and need to be monitored for risk factors. PMID- 17405292 TI - [Erythropoietin prevents oxidative stress and lipid peroxidation process in human umbilical vein endothelial cells induced by tumor necrosis factor alpha]. AB - According last research erythropoietin (Epo), glycoprotein known as a physiological regulator of erythropoiesis and used successfully to treat anemia, displays cytoprotective properties. It is supposed that the protective effect of EPO is based on its ability to oxidation-reduction stabilization of cells. AIM: The aim of the study was to examine the influence of oxidative stress and lipid peroxidation in human umbilical vein endothelial cells (HUVEC) induced by tumor necrosis factor-alpha (TNF-alpha). MATERIAL AND METHODS: Human umbilical vein endothelial cells were cultured in a standard medium (M 199). The experiment was performed five times on the forth passage of HUVEC culture. For stimulation TNF alpha was used in concentrations: 10, 20, 40 ng/ml (6 hours) and erythropoietin in concentrations: 10, 20, 40 IU/ml (24 hours). In the HUVEC lysate malonyldialdehyde (MDA), lipid hydroperoxides and reduction gluthation (GSH) concentrations were measured by HPLC method. RESULTS: MDA concentration statistically significantly decreased from 16.48 +/- 1.21 to 14.40 +/- 0.72 nmol/mg protein and LOOH concentration from 73.00 +/- 5.44 do 68.86 +/- 1.89 nmol/mg protein in the cell's lysate due to the preincubation with Epo of HUVEC cells, stimulated with TNF-alpha. MDA and LOOH concentrations in the control culture were 12.91 +/- 1.02 nmol/mg protein and 57.80 +/- 6.16 nmol/mg protein respectively. The application of erythropoietin while the cells were being stimulated with TNF-alpha prevented the decrease in GSH concentration which was 34.77 +/- 0.70 nmol/mg protein in the control culture, 33.11 +/- 1.65 nmol/mg protein in the culture stimulated with TNF-alpha and 34.17 +/- 0.14 nmol/mg protein in culture preincubated with Epo and stimulated by TNF-alpha. CONCLUSIONS: Erythropoietin (Epo) prevents oxidative stress and lipid peroxidation process in human umbilical vein endothelial cells (HUVEC) induced by tumor necrosis factor-alpha. PMID- 17405293 TI - [Computed tomography assessment of incorporation bone grafts after cemented acetabular revision]. AB - The aim of the study was assessment, on the basis of computed tomography (CT) examination, of remodelling of compacted frozen bone sliver grafts used in acetabular revision arthroplasty. MATERIAL AND METHOD: The study involved ten patients treated in the years 1998-2000 for aseptic loosening of hip joint prosthesis elements. Eight women and two men were studied. The mean age of the patients was 69.6 years (from 52 to 74 years). The mean time between primary and revision arthroplasty was 6.8 years. Spiral CT examination was performed 6 weeks and 3, 6, 12, 24, 36 months after the operation. Density measurement was done using ROI (region of interest) function including spongy bone area from 50 to 80 mm2. The studied layer thickness was 3.2 mm. In the examination both hip joints were assessed, accepting the joint not operated on as standard for a given patient. The measurement was done in six zones according to DeLee, Charnley. RESULTS: Progressing reduction of graft density was found in relation to healthy bone density. During the first three months most rapid density reduction was observed in all patients. The lowest density decrease was seen in the postero superior part of the acetabulum while the highest density reduction occurred in the lower part. Trabeculation change was observed from that resembling compact bony tissue to that corresponding to spongy tissue. In two patients after six months a significant graft density reduction occurred both in the lower and upper parts of the acetabulum. In these patients acetabular loosening was confirmed, requiring reoperation. CONCLUSIONS: CT examination of bone density makes possible quantitative assessment of bone graft remodelling and qualitative assessment imaging the structure of the grafts. The non-invasive method of CT examination of bone graft density allows for prognostication as to stability of the seated acetabulum. PMID- 17405294 TI - [Bone mineral density in children and adolescents with idiopathic chronic musculoskeletal pain syndromes]. AB - PURPOSE: There is a limited information regarding associations between bone mineral density (BMD) and idiopathic musculoskeletal pain in pediatric subjects. The aim of this cross-sectional study was to assess whether children and adolescents with pain syndrome have lower BMD than healthy subjects. MATERIAL AND METHODS: Eighty-nine subjects (49 girls, 40 boys) aged 5-18 years with chronic non-rheumatic musculoskeletal pain and daily calcium consumption below 500 mg were involved in the study. The subjects were divided in three groups: I- prepubertal (5-9 years), II--pubertal (9-15 years) and III--adolescents (15-18 years). Bone mineral density in the total skeleton (total BMD) and lumbar spine (spine BMD) was examined using dual energy x-ray absorptiometry (DXA) and compared to reference data. Serum calcium, ionized calcium, phosphate, alkaline phosphatase and its bone-fraction were estimated. RESULTS: Low BMD (below the 5th percentile) was found in about 50% of the participants with pain syndromes in each measurement site. A significantly decreased Spine BMD was observed in those children who reported pain symptoms in the spine region and lower limbs (Z-score for Spine BMD was = -1.55; -1.41) compared to subjects with non-localized pain. The lowest mean Z-score (-1.85) for Spine BMD was found in the postpubertal group (III) compared with groups I and II. CONCLUSIONS: As one-half of children and adolescents reporting musculoskeletal pain had low BMD it is possible that the symptoms are associated with an inadequate bone mass accrual during growth. Pain localized in the spine region and in lower extremities may be a selective and site-specific symptom of juvenile osteoporosis without fractures. The chronic musculoskeletal pain suggests a rationale for bone density testing during growth. PMID- 17405295 TI - [Analysis of loss of heterozygosity and microsatellite instability RAD52, RAD54 and RAD54B gene and BRCA1 gene mutation in breast cancer]. AB - RAD51, RAD52, and RAD54 encode proteins that are critical to the repair of double strand DNA breaks by homologous recombination. The objectives of this study were to determine the frequency of BRCA1 germ-line mutations and the RAD52, RAD54 and RAD54B microsatellite instability (MSI) and loss of heterozygosity (LOH) in patients with breast cancer. MATERIAL AND METHODS: Blood and tumour tissue were obtained from 100 breast cancer women. Blood samples age matched healthy individuals (n = 60) served as control. RESULTS: The microsatellite instability status was significantly higher in breast cancer tissue compared as control (p < 0.05). Significant correlation between one or more regions with concomitant LOH and pathologic parameters were observed with respect to progesterone receptors (p = 0.004) and tumour grade (p = 0.001). In present study one Ex20insC and two ExII17delA mutations of BRCA1 gene were identified in women with breast cancer. CONCLUSION: The results suggest that the microsatellite instability and LOH seems to be important in the development sporadic 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- 17405296 TI - [Composition of linoleic and arachidonic acids in phosphatidylcholine of erythrocytes membrane and IL-6 and TNF-alpha in serum and C-reactive protein concentration in children with juvenile idiopathic arthritis]. AB - The study objective was to determine the relationship, if any, between the levels of arachidonic and linoleic acids in erythrocyte membrane phosphatidylcholine (PCH), serum interleukin 6 (IL-6) and TNFalpha levels and C-reactive protein (CRP) in juvenile idiopathic arthritis (JIA). MATERIALS AND METHODS: The study involved 49 children with JIA, aged 3-18 years (mean 11.3 +/- 3.9), and 29 healthy subjects. The JIA children were divided into 2 groups: group I--24 children (in exacerbation period) with mean CRP level of 15.6 mg/L +/- 13.3 and group II--25 children (in remission period--joint swelling-free) with CRP of 7.8 mg/L +/- 5.8 on average. Lipids were extracted according to a modified Folch's method. Fatty acids in erythrocyte membrane PCH were identified using gas chromatography (Hewlett-Packard 5890). The levels of IL-6 and TNFalpha were determined by ELISA, using Quantikine sets: R&D System (USA), while CRP was measured by nephelometric method on a BN II apparatus (Behring). RESULTS: We found a significant decrease in the level of linoleic acid (p < 0.05) and a statistically insignificant reduction in arachidonic acid in JIA patients as compared to the controls. The decrease in linoleic acid was more pronounced in the active phase of JIA (p < 0.001. The higher serum CRP level was accompanied by a significantly elevated level of IL 6 (p < 0.05). The concentration of TNFa was elevated, but the difference had no statistical significance. CONCLUSIONS: The levels of linoleic and arachidonic acids in erythrocyte PCH decreased and the concentrations of IL-6 and TNF-alpha increased in JIA children in the active phase of the disease. The differences intensified with a rise in CRP. PMID- 17405297 TI - [Beneficial actions of lipoic acid in cerebral cortex homogenates in rats during lipopolysaccharide-induced oxidative stress]. AB - The aim of his study was to assess the effect of lipoic acid (LA) on lipid peroxidation, hydrogen peroxide(H2O2) concentration, sulfhryl group (-SH) contents in the cerebral cortex homogenates in the early stage of oxidative stress. MATERIALS AND METHODS: Experiments were performed on male rats Wistar 300 320 g body weight. The animals were divided into four groups consisting of eight animals each. Control group received saline alone into the tail vein (i.v.). The second group received LA (60 mg/kg b.w., i.v.). The third group received LPS (30 mg/kg b.w., i.v.). The fourth group received LPS (30 mg/kg b.w., i.v.) and after 0.5 h received LA (60 mg/kg b.w., i.v.). In the homogenates from cerebral cortex were measured thiobarbituric acid reactants (TBARS) as indicator lipid peroxidation, H2O2 concentration and -SH groups contents. RESULTS: The administration of LA after LPS-induced oxidative stress caused a significant decrease in TBARS and H2O2 concentration 5.38 + 0.084 micromol and 0.028 + 0.002 micromol, respectively) compared with the group treated with LPS (6.96 + 0.063 micromol and 0.198 + 0.003 micromol, respectively). Treatment of LPS-injected rats with LA caused increase in -SH groups (p < 0.05). CONCLUSION: The early administration of LA a significant suppressed symptoms of oxidative stress induced LPS, expressed as a decrease in lipid peroxidation process, H2O2 generation and increase in concentration of protein containing thiol groups in the cerebral cortex homogenates. PMID- 17405298 TI - [Specific immunotherapy in the treatment of patients with atopic dermatitis- results of double blind placebo controlled study]. AB - BACKGROUND: Specific immunotherapy (SIT) is the only causative method of treatment in the case of IgE-mediated allergic disease. There is extensive literature on the use of SIT in the management of respiratory allergies and insect venom hypersensitivity, but a paucity of published data concerning its use as a therapeutic approach in atopic dermatitis (AD). OBJECTIVE: A double blind placebo controlled trial was conducted over a period of 12 months in order to evaluate the efficacy of SIT in the management of atopic dermatitis attributable to house dust mites or grass pollens. METHODS: A total of 20 patients (5-40 years) with AD and monovalent sensitization to airborne allergens (house dust mites or grass pollens) were enrolled in the study. SIT was performed using aluminium hydroxide adsorbed allergen preparations administered by subcutaneous injection. Clinical efficacy of the treatment was assessed using the clinical score--W-AZS index. Serum concentration of total IgE and allergen specific IgE were measured, as were various immunological parameters including ECP, sIL-2R, IFN-gamma, IL-4, IL-5 and IL-10). RESULTS: The mean value of W-AZS index in the SIT group before treatment was 87.6 +/- 15.8 pts, and this decreased to 38.8 +/- 34.4 pts after 12 months of therapy (p < 0.01). In the placebo group the mean W AZS index before treatment was 86.3 +/- 15.7 pts and after 12 months of therapy it increased to 111.9 +/- 41.7 pts. Comparative statistical analysis indicated a significant difference between the two groups in favour of patients treated with the active allergy vaccines (p < 0.01). Serum levels of specific IgE in the SIT group showed a tendency to decrease, whilst those in the placebo group tended to increase. Serum concentrations of selected immunological parameters including ECP, sIL-2R, IFN-gamma, IL-4, IL-5 and IL-10 were monitored before and after treatment, but did not show significant differences. CONCLUSION: Allergen specific immunotherapy appeared to be an effective method of treatment for atopic dermatitis as judged by significant improvement in clinical index in cases with well documented IgE-mediated allergic disease. PMID- 17405299 TI - [Depression and pain in arthrosis of spine and hip in elderly patients]. AB - Various pain complaints are the most frequent symptoms in depression. However neither ICD-10 nor DSM-IV do not treat the pain as an essential and vital indicator of depression. There is also lack of works which describe shown above relation. AIM: The aim of our research determines the connection between the intensity of depression symptoms and pain perception in elderly patients who are treated for pain in arthrosis of spine and arthrosis of hip. MATERIAL AND METHODS: The research was conducted in a group of 60 patients. The whole group was divided into two subgroups: group A (30 people with arthrosis of spine: 22 women and 8 men) and group B (30 people with arthrosis of hip: 23 women and 7 men). All of them were undergone 10 physiotherapy surgeries (magnetic field and therapy laser). The appraisal of the influence of the treatment on the intensity of depression symptoms was made on the basis of Depression Scale Montgomery Asberg (MADRAS) and the appraisal of the intensity of pain was made on the basis of Visual Analogy Scale of Pain. RESULTS: The important impact of conducted surgeries on the intensity of pain complaint as well as the escalation of depression indicators were stated. Not only in the first but also in the second case presented patients' diseases were alleviated (the intensity of depression symptoms and of perceptible suffering decreased). Nevertheless, this relation was observed only when moderate and mild escalation of depression appeared. CONCLUSIONS: The decrease on the intensity of pain experience reduce depression symptoms in these cases in which the escalation of the second ones is not substantial. PMID- 17405300 TI - [Mastocytosis--mastcell proliferative disease risk of anaphylactic reaction]. AB - Mastocytosis is a group of rare diseases characterized by abnormal growth of mast cells in skin, bone marrow, liver, spleen, lymph nodes. Signs and symptoms result mostly from mast cells mediators and mast cells organ infiltration. Pathological examination proving mast cells infiltration is crucial for the diagnosis of disease. Therapy covers patient education and symptomatic treatment (antihistamine drugs and glicocortycoids). Attempts of interferon, cladribine, imatinib treatment are made. Aggressive forms of diseases require, chemiotherapy, bone marrow transplantation. All mastocytosis subjects should be equipped in adrenaline. The paper describes case of 52 years old woman who suffered from urticaria pigmenthosa, anaphylaxis, insect venom and food allergy. Diagnosis included bone marrow examinations (pathology, cytology, genetics, cytofotometry) tryptase level, skin prick tests and sIgE. Mastocytosis was diagnosed. Therapy included symptomatic treatment and immunotherapy. The paper describes also aims of the European Competence Network on Mastocytosis. PMID- 17405301 TI - [Subclinical thyroid disorders]. AB - The term subclinical thyroid disorders apply to patients who have an abnormal serum thyroid-stimulating hormone (TSH) concentration but thyroxine and triiodothyronine levels within their reference ranges. Subclinical hypothyroidism occurs in 4% to 10% of the general population, and is especially prevalent in elderly women. Depending on the study, subclinical hyperthyroidism has been found in 0.6-16 % of the population. Potential risks of subclinical hypothyroidism include progression to overt hypothyroidism, dyslipidemia, cardiovascular complications, and neurological and neuropsychiatric effects. In turn, subclinical hyperthyroidism represents a considerable risk factor for atrial fibrillation in the elderly and for postmenopausal osteoporosis. Treatment of subclinical thyroid disorders is controversial. In this paper, we discuss the prevalence, aetiology, clinical presentation, diagnosis, natural history, and potential pathophysiological consequences of subclinical hypothyroidism and subclinical hyperthyroidism. We also provide some screening and management recommendations. PMID- 17405302 TI - [Hyperthyroidism in pregnancy]. AB - Thyroid disorders constitute one of the commonest endocrine problems found in pregnant women. They are usually treatable but may affect mother and foetus adversely if they are not evaluated relatively early and managed appropriately. The diagnosis of hyperthyroidism in pregnancy may be often challenging due to the fact that its symptoms mimic many of normal pregnancy complaints. Moreover, some forms of hyperthyroidism occur exclusively during this period. The goal of this article is to summarize the present state of knowledge on the etiology, clinical presentation, recognition and management of the various types of hyperthyroidism in pregnancy. On the basis of numerous studies, carried out mainly in the recent years, we provide some practical guidelines for clinical endocrinologists dealing with treatment of pregnant women. PMID- 17405303 TI - [Use of transcutaneous electrogastrography in the diagnostics of digestive tract diseases]. AB - Transcutaneous electrogastrography (EGG) is the only non-invasive method of gastric myoelectric activity assessment which allows to evaluate slow wave activity and peak potentials of gastric contractions. During single channel analysis dominant frequency of slow waves (DF) is assessed, dominant power (DP), difference of DF and DP before and after the meal, percentage of normo-, brady- and tachygastry, ratio of dominant postprandial activity to preprandial acivity (power ratio--PR) and dominant frequency instability coefficient--DFIC). Multichannel EGG let us to assess myoelectric activity of different parts of stomach during one analysis. Transcutaneous electogastrography allows only for indirect evaluation of motoric disorders and needs many more studies to more precise assessment of its clinical use in gastroenterological diagnostics. PMID- 17405304 TI - [Cognitive functions and diabetes]. AB - In the presented work, the researches of foreign and Polish authors were reviewed. The aim of this project was to find the answer to the question connected with the character and the cause of diabetes' influence on cognitive functions of patients. Diabetes is a metabolic condition of a various etiologyaetiology. It is distinguished by chronic hiperglycemia with the disorder of metabolism of carbohydrates, fat and proteins as a result of giving off defects and/or working insulin. Cognitive functions are these psychological functions which the human needs in order to be knowledgeable about surroundings, to get the information about oneself and your body, to analyse the situations, to formulate conclusions, to take a right decisions or to function. In the last few years, this typical somatic disease, as the diabetes is, puts pressure on cognitive efficiency of the human body. The vita issue is to get to know this problem not only from medical or psychological point of view but also it has a great influence on patients' better life. The knowledge of this subject should be known among medical surroundings as well as in all the places where the patients live (school, workplace and family). PMID- 17405305 TI - [Treatment of clinically localized prostate cancer. Part I--observation or treatment?]. AB - The optimal management of clinically localized (T1, T2) prostate cancer remains controversial. In response to these controversies the American Urological Association guidelines considered surgery, radiotherapy or watchful waiting to be options, because the literature does not provide clear evidence for the superiority of any one treatment. The aim of this paper is presentation of current knowledge regarding this problem. PMID- 17405306 TI - [Treatment of clinically localized prostate cancer. Part II--efficiency comparison of therapy methods]. AB - The optimal management of clinically localized (T1, T2) prostate cancer remains controversial. In response to these controversies the American Urological Association guidelines considered surgery, radiotherapy or watchful waiting to be options, because the literature does not provide clear evidence for the superiority of any one treatment. The aim of this paper is presentation of current knowledge regarding this problem. PMID- 17405307 TI - [Treatment of clinically localized prostate cancer. Part III--external beam radiotherapy]. AB - The optimal management of clinically localized (T1, T2) prostate cancer remains controversial. Patients have possibility of choice between prostatectomy and radiotherapy in two forms: external beam radiotherapy and brachytherapy. Multicentre studies show comparable results of theise two methods. The aim of this paper is to present current knowledge regarding treatment with conformal radiotherapy. Acute and late effects of ionizing radiation are described. Propriety of associate radiotherapy with hormonotherapy was analyzsed. PMID- 17405308 TI - Experimental studies on bacterial product cantastim derived from Pseudomonas aeruginosa. VII. Activation of immune cells of healthy controls and cancer patients. AB - CANTASTIM (CS) is a purified extract of Pseudomonas aeruginosa with beneficial effects related to enhancing the immune responses in conditions such as chronic viral and bacterial infections, immunodeficiencies and cancer immunosuppression. The aim of this study was to determine the capacity of this biological product to stimulate in vitro human leukocytes in whole blood. Blood samples from healthy donors and cancer patients were incubated with CS for 24 h and leukocytes were assessed for induction of inflammatory cytokines (TNF-alpha and IL-1beta) by ELISA and expression of early activation marker CD69 by flow-cytometry. For both groups of investigated subjects, the levels of TNF-alpha and IL-1beta in the supernatants of whole blood culture stimulated with CS were significantly higher than in unstimulated cultures, although lower than in LPS-stimulated samples. Stimulation of whole blood cultures with CS increased both the frequency and the expression of CD69 on the surface of T lymphocytes and NK cells. Importantly, this was noticed not only for healthy controls, but also for cancer patients. These data demonstrate the capacity of bacterial immunomodulator CS to activate human leukocytes of healthy subjects and cancer patients. PMID- 17405309 TI - Microbiological and immunological study of staphylococcus vaccine effects in periodontitis. AB - The aim of this study was to evaluate the immunomodulatory effect of the staphylococcal vaccine inoculated subcutaneously in 15 patients with chronic periodontitis. Bacteriological investigation of samples collected from the periodontal pocket for aerobic and anaerobic microorganisms was performed by classic bacteriological procedures before and after vaccination. The following immune system parameters were evaluated: C reactive protein (CRP), serum level of C3 complement fraction, IgG, IgA, and IgM by immunodiffusion, PMN granulocytes ROS release after in vitro stimulation with opsonized zymosan (OZ) and Concanavalin A (ConA) by chemiluminescence assay and lymphocytes sets and subsets by flow-cytometry immunophenotyping. The microbiological investigations revealed high frequency of Staphylococcus spp isolation and the presence of the most common anaerobe agents incriminated in human periodontitis like Fusobacterium, Porphyromonas, Peptostreptococcus, Veillonella spp and the reduction of this flora in the periodontal pocket after therapy. The immunological parameters quantification showed the absence of CRP, normal values of C3, IgG, IgA, IgM in the majority of cases. All patients presented normal values of lymphocytes sets and subsets. Significant increase of PMN respiratory burst after ConA stimulation was observed before vaccination which turned to normal values after therapy and a low ROS level both before and after therapy suggesting PMN Fc receptors dysfunction in this group of patients. The data presented in our study suggest an immunomodulatory effect of staphylococcal vaccine therapy in periodontitis and high frequency of Staphylococcus spp recovering from the periodontal pocket of investigated subjects. PMID- 17405310 TI - Patterns of peripheral cellular immune disorders in severe rheumatoid arthritis. AB - The study is focused on the correlated peripheral cellular immune disorders registered in a group of 23 patients with severe, progressive rheumatoid arthritis, on methotrexate therapy. We investigated a panel of peripheral immune parameters: leukocyte counts, the proportions of lymphocyte populations (T, Thelper, Tcytotoxic/suppressor, B lymphocytes and NK cells) and the polyclonal activation of lymphocytes. Results show that leukocytosis is due to simultaneously elevated values of monocytes, granulocytes and, to a lesser extent, lymphocytes. The registered high values of the Th to Tc/s ratio are mainly attributed to the abnormal low proportions of the Tc/s subpopulation. Inverse correlations were emphasized between B, Tc/s lymphocytes and NK cells or granulocytes. The unbalance of the lymphocyte to monocyte ratio or of the Th to Tc/s ratio does not impair the polyclonal activation of lymphocytes. In conclusion, we have characterized different patterns of correlated cellular peripheral immune disorders in rheumatoid arthritis, associated to pathological processes in conjunction with the immunsuppressive and anti-inflammatory action of methotrexate that might be relevant for further investigation of disease and further therapy outcome. We emphasize the special relation between the adaptive and innate immune system at the level of cell counts and proportions. The correlations between the peripheral abnormalities in the rheumatoid arthritis group are better highlighted by analyzing subgroups of patients characterized by particular values of the investigated parameters. PMID- 17405311 TI - Neurotrophin-3, TNF-alpha and IL-6 relations in serum and cerebrospinal fluid of ischemic stroke patients. AB - Pro-inflammatory cytokines and neurotrophins in the central nervous system (CNS) have been recognized as mediators of both neurodegenerative and neuroprotective mechanisms in a number of CNS pathologies. A rapid, sustained elevation of these molecules was recently reported after traumatic and ischemic brain injury. Inflammatory mechanisms and immune activation have been hypothesized to play a role in the pathogenesis of cerebral ischemia. Stroke is the third largest cause of death next to heart disease and cancer in the world, and it is an important cause of death and disability in developed countries. Role of excitatory amino acids receptors activation, calcium overload, nitric oxide and oxidative stress in the pathogenesis of ischemic brain damage is well established. Stroke may modulate peripheral neurotrophic factors levels. In experimental animal models, neurotrophin-3 (NT-3) has been shown to be produced by glial cells as an adaptability response to hypoxia. In spite of substantial research and significant number of neuroprotective drugs that have been developed to limit ischemic brain damage and to improve the outcome for stroke patients, no specific therapy for stroke is available. The neurotrophins have been proposed as therapeutic agents for the treatment of neurodegenerative disorders and ischemic injury. In the present work, we investigated the possible correlation of NT-3 with tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in the serum and cerebrospinal fluid (CSF) from patients with ischemic stroke (IS). PMID- 17405312 TI - Incidence of virulence-encoding genes among enteric Escherichia coli strains isolated from healthy subjects. AB - Escherichia coli, heterogeneous species consisting of commensal and pathogenic strains, is causing a broad spectrum of intestinal and extra intestinal diseases, ranging from asymptomatic infections to septicaemia, according to its capacity to produce different virulence factors. The incidence of different virulence associated genes among the strains isolated from healthy subjects, taking into account that the human gastrointestinal tract is considered an important source for spreading E. coli strains, was evaluated. A total of 241 E. coli strains isolated from 41 healthy subjects, working in the food chain and coming to the laboratory for periodical medical control, were investigated for harbouring patogenicity factors--encoding genes. Extra intestinal virulence-associated genes, pap, sfa/foc, afa, hly, cnf and intestinal ones eaea, bfp, agg, It, st, vtx1 (stx1), vtx2 (stx2) and ipaH, were targeted by PCR using cellular lysate for total DNA. Genes encoding for adherence were the most prevalent. A number of 67 strains (27.80%) were positive for pap genes and 34 strains (14.11%) presented PCR positive results when afa genes were targeted, but sfa/foc genes were identified in only 10 strains (4.15%). Genes encoding for toxigenesis were less prevalent. A total of 9 strains amplified hly genes, 2.49% were positive for cnf genes and only 2 strains presented vtx1(stx1) gene. The results are in concordance with those which demonstrate that healthy subjects carrying strains possessing virulence-encoding genes could represent a reservoir for environmental circulation of such strains, considered life-threatening when a receptive host is encountered. PMID- 17405313 TI - Application of pulsed-field gel electrophoresis to laboratory surveillance of small community outbreaks of Salmonella enterica serovar Typhimurium. AB - Infectious diarrhea syndrome is an important cause of human morbidity around the world, and Salmonella genus remains one of the most prevalent etiology. Salmonella enterica serovar Typhimurium outbreak-associated isolates received by the Laboratory for Enteric Pathogens from N.I.R.D.M.I. "Cantacuzino" for confirmation and typing were analyzed by genomic pulsed-field gel electrophoresis (PFGE) and phage susceptibility testing to establish their relatedness. Both typing methods proved to have similar discriminatory power. The isolates originating from the same outbreak belonged to the same phage type and showed indistinguishable PFGE profiles. The molecular characterization of autochthonal Salmonella enterica Typhimurium outbreak human isolates provided laboratory evidence that epidemiologically related isolates collected from community outbreaks of disease were also genetically related. In order to improve the national and international surveillance of major foodborne pathogens the reference laboratory centers are required to establish and maintain the capacity to perform a wide range of both phenotypic and genotypic methods to support outbreak investigations. PMID- 17405314 TI - Rapid immunochromatographic serum assay of nontuberculous mycobacterial infections. AB - A rapid immunochromatographic serologic assay (Dot assay) is proposed to be applied on patients infected with nontuberculous mycobacteria (NTM). This assay could evidentiate the infecting species and allow the beginning of the treatment. The test is based on the principle of immunoblotting chromatography, a rapid membrane-based assay, capable of diagnosing NTM infections in serum, in less than 1 hour, with no need of special equipment or skilled staff. The secreted extracellular antigens have been isolated from the unheated culture filtrates of the clinically significant NTM (M. avium, MAI, M. kansasii, M. xenopi, M. chelonaei, M. scrofulaceum, M. marinum, M. fortuitum, M. abscesus, M. szulgai). The patients have been tested against these antigens, as well as from M. tuberculosis H37Rv, due to the possibility of co-infection with tuberculous bacilli. A number of 385 tests on patient sera have been performed (10, with NTM suspected infection, with or without M. tuberculosis co-infection, 5 with confirmed diagnosis of NTM infection, 10 with TB, 10 with other respiratory diseases). The preliminary results presented in this paper support the fact that the rapid immunochromatographic serum assay, combined with clinical and radiographic evidence, could evidentiate the infecting NTM species and allow the start of an earlier treatment, but must be confirmed on a higher number of patients. PMID- 17405315 TI - Intracellular degradation of poly(3-hydroxybutyric acid) accumulated by Azotobacter chroococcum MAL-201. AB - Azotobacter chroococcum MAL-201 accumulates poly(3-hydroxybutyric acid) [P(3HB)] accounting 69% of cell dry weight (CDW) from glucose during growth in nitrogen free Stockdale medium. Degradation of the accumulated polymer by the organism was studied under carbon-free medium following two-step cultivation method. P(3HB) content of cells decreased rapidly from 69% to 4.8% of CDW after 35 h under carbon-deprived condition. Autodigestion of P(3HB) was evident from the estimation of intracellular P(3HB) depolymerase (i-depolymerase) activity in cell free extract using artificial P(3HB) granules as substrate. Polymer content decreased rapidly along with the increase in i-depolymerase activity and rate of polymer degradation when medium was supplemented with (NH4)2SO4 at 0.1% (w/v) level. However, the effects were reverse when organic nitrogenous substrate, beef extract at similar concentration was present in the medium. The optimum temperature and pH for i-depolymerase activity were 35 degrees C and 7.7 respectively. The oxygen-limiting condition (culture volume per flask volume, 50%) decreased 10.7% activity of i-depolymerase over control resulting a slow P(3HB) degradation. The presence of NaCl (6 x 10(3) microg/ml) showed a positive effect on i-depolymerase whereas EDTA (40 microg/ml) resulted in 20% less activity. Furthermore, the intracellular degradation of P(3HB) decreased the intrinsic viscosity, molecular weight and tensile strength of the accumulated polymer. PMID- 17405316 TI - The effect of the plasma needle on tumoral cell lines apoptosis. AB - In this paper, we present the effect of the plasma needle on tumor cell surface. The plasma is generated at the tip of a metal needle by using a radio-frequency generator of 13.56 MHz, 100's V amplitude. In our study we investigated the interaction of non-thermal plasma (plasma needle) with living monolayer tumour cells in culture medium. We applied short needle to sample distance (1 mm) at temperature of 25 degrees C, 30 degrees C and 37 degrees C, respectively. Our data sugest that the plasma needle reduces the viability and induces apoptosis of tumour cells. These activities may be very useful in dermatology, where a part of the tissue must be removed with high-precision, without damage to the adjacent cells and without inflammatory reaction. PMID- 17405317 TI - Preliminary identification and classification of five new yeast strains isolated from oil-polluted environment. AB - Bioremediation is a very interesting alternative for restoring the oil-polluted ecosystems. Many studies concerning the possibility of using microorganisms (bacteria and yeasts) in the degradation of oil compounds have as starting point the isolation and taxonomical identification of new species and strains with degradative abilities. Our study focusses on the preliminary classification of five yeast strains (D1, D2, D3, D4 and D6) isolated from oil-polluted environments. The strains were characterized by conventional taxonomical techniques: microscopical and macroscopical appearance, fermentation abilities, assimilation of various carbon or nitrogen compounds, growth under stress conditions (non-permissive temperatures, high glucose concentration) and urea degradation. According to these tests, D1, D2 and D4 showed great similarity to Rhodotorula glutinis, D3 to Candida parapsilosis and D6 to Candida tropicalis. Further supplementary tests were performed in order to establish their ability to degrade hydrocarbons, by observing growth in media with n-alkanes (n-decane, n dodecane, n-tetradecane, n-hexadecane). Thus, D1, D2 and D4 were the best alkane consuming strains, presenting possible similar degrading abilities and pathways, which correlates well to our identification as Rhodotorula strains. For D3 and D6 the growth was not so spectacular as for D1, D2 and D4, but continuous along the entire experiment. The resemblance between the curves profiles confirms the idea that both belong to the same genus, Candida. PMID- 17405318 TI - Case-control study to evaluate the link between immunosuppression and Candida spp. infection. AB - Candidiasis is one of the fungal infections with the highest incidence in the immunosuppressed host. The evolution of infection and the increase of antifungal medical drugs resistance could both contribute to the mortality attributable to Candida infection in the immunosuppressed host. Even though the data from international studies are well known, few studies have been published in Romania on this subject. In the case-control study we demonstrated the link between the immunosuppression and the presence of Candida infection. Further studies are to be carried out in order to identify more accurately this link and to extend the study to other fungi. There is a need to increase the microbiological diagnosis use at least at the hospital laboratory level in order to better identify the real situation of fungal infections and the link between them and the concrete status of different hosts. Continued surveillance for infections caused by C. albicans and other species of Candida among hospitalized patients is recommended. Control of antimicrobial resistance among nosocomial infections caused by C. albicans and other species of Candida requires rational policies for use of both antifungal and antibacterial agents and appropriate surveillance for the emergence of resistant strains and species. PMID- 17405319 TI - A study on physicochemical parameters of an aquaculture body in Mysore city, Karnataka, India. AB - Monthly changes in water quality parameters (physicochemical) of a rain fed lake (Bilikere) in Mysore city, were investigated for two calendar years (2002 and 2003) to assess the suitability of this lake for pisciculture. Although there were monthly fluctuations in water temperature, total suspended solids (TSS), dissolved oxygen (DO), nitrite and ammonia, they were within the desirable limits. On the other hand, total alkalinity and hydrogen sulphide throughout the study period and pH for a major part, were higher than the desirable limits. Other parameters viz; turbidity, biological oxygen demand (BOD), phosphate, and nitrate in a few months were higher than the desirable limits for waters used for fish culture. The high levels of these factors are due to the entry of agricultural run off and occasional flow of sewage into the lake. In addition dense algal growth was noticed at times of the year which is caused by surge in nutrients level whenever there was a rainfall. Since, the lake has a great aquacultural potential, it is suggested that control of nutrient load that enters the lake occasionally, might help the lake to continue its mesotrophic status. PMID- 17405320 TI - Responses of the whipscorpion, Mastigoproctus liochirus (Arachnida, Uropygi) to environmental humidity. AB - Experiments were conducted to determine the responses of third-instar nymphs of the whipscorpion Mastigoproctus liochirus to various moisture (relative humidity) levels under constant temperature conditions, using a linear humidity gradient apparatus. No previous data exist on the water relations of this uropygid. Under saturated conditions (100% RH) animals showed no preference for any section of the chamber. When a humidity gradient was established (20 to 100% RH), whipscorpions exhibited a marked preference for an area of the chamber characterized by 70 to 80% RH. PMID- 17405321 TI - A comparative study on the toxic effects of textile dye wastewaters (untreated and treated) on mortality and RBC of a freshwater fish Gambusia affinis (Baird and Gerard). AB - Comparative toxicological studies of textile dye wastewater (untreated and treated) on a freshwater fish, Gambusia affinis, revealed a marked reduction in mortality and cytotoxic effects on RBCs, measured as reduction in their counts and percent changes in their shape (poikilocytosis) and variation in their size (anisocytosis)}, after subjecting them to both physicochemical and biological treatments. On comparing the data of mortality and the cytotoxic effects on RBCs, we found poikilocytosis is a better indicator for toxicity measurement of both untreated as well as treated wastewater, especially at their lowest concentrations where percent mortality was found to be either nil or lowerer than the percentage of poikilocytic RBCs. Although percent reduction in RBC counts and changes in their size (anisocytosis) indicated toxic effects of wastewaters, but EC5o values for RBC counts were usually higher than those for poikilocytosis and mortality, and non-calculable for anisocytosis suggesting their lesser sensitivity to pollutants. In view of these findings, we recommend monitoring of toxic effects of wastewaters during fish bioassay on both mortality and variation in RBC shape. PMID- 17405322 TI - Reduction of the impact produced by sea lions on the fisheries in Mexico. AB - Activities of fishermen in the Bay of La Paz, B. C. S. are focused to satisfy the local demand of fish and shellfish by using approximately 300 small outboard crafts equipped with gillnets. Sea lions in this Bay attack the fishes captured damaging both product and gear. We did experimental gillnet throws to determine the frequency and preferences of sea lions in fishing areas. 52 experimental gillnet throws with time averages of 2 hr were performed, rending an average of 30 kg of fish captured and less than 10% of damages to the net.. Traditional fishermen in this Bay usually left the net the whole night, (approximately 7.50 hr, obtaining an average of 50 kg of captured fish, but the damages to the nets is in average of 40%. The cost-benefit balance comparing our alternative fishing method, which includes the use of the gillnets during the afternoon, watching for sea lions and retiring the nets at their arrival, it is more sustainable and profitable than the traditional fishing method currently used by the local fishermen. This paper suggests how to minimize the harmful effects of the sea lions on the fishermen productivity and gear, maximizing the production and reducing the damage. Our alternative method is applicable to other regions where this harmful interaction is taking place. We conclude that the coexistence of sea lions-fisheries is feasible, by applying the simple measures that we propose. PMID- 17405323 TI - Groundwater quality--assessment on Anekal Taluk, Bangalore Urban district, India. AB - Water as a resource, basic amenity and universal solvent is shared by population. The physicochemical and biological quality study of Anekal Taluk has been taken up to evaluate its suitability for potable purposes. 1026 water samples were collected from bore well, hand pumps of 272 villages covering in Anekal Taluk. The quality of groundwater has been made through the analysis of pH, colour, electrical conductivity, turbidity, total dissolved solids, alkalinity, chlorides, total hardness, calcium, fluoride, nitrate, sulphate, iron and E. coli. The quality of groundwater assessed in the study area is discussed in detail. PMID- 17405324 TI - Biosorption of Cu2+ from aqueous solutions by pretreated Cladosporium sp. AB - In this paper, adsorption properties of a pretreated of Cladosporium sp. for Cu2 were studied. The fungi pretreated with some chemical reagents exhibited higher Cu2+ removal capacities than native biomass. The optimum chemical reagent was 0.2M NaOH. After 0.2M NaOH pretreatment, optimum conditions of biosorption were found to be pH 5.0, temperature 35 degrees C, and stirring speed of 100rpm. Equilibrium isotherms were obtained from adsorption experiments and the biosorption maximun capacity obtained was at 28.31mg/g. The biosorbed metal ions were effectively eluted by 0.05M HNO3 solution. After eluting, the biosorbed metal ions biomass was regenerated by washing with deionized water and then contacted with a solution containing 0. 1M of Ca2+, Mg2+ ions before further adsorption tests. The pretreated fungi biomass could be used for three cycles: biosorption, elution of biosorbed ion, regeneration of biomass. PMID- 17405325 TI - Toxicity and bioaccumulation of cadmium and lead in Salvinia cucullata. AB - The toxicity and accumulation of heavy metals, cadmium (Cd) and lead (Pb) in aquatic fern, Salvinia cucullata were studied. Plants were cultured in Hoagland's medium which was supplemented with 0.5,1,2, and 4 mg/l of Cd and 5, 10, and 40 mg/l of Pb and were separately harvested after 2,4,6, and 8 days. The toxicity symptoms of Cd and Pb to S. cucullata showed chlorosis on leaves. There were significant derceases in the relative growth, biomass productivity and total chlorophyll content when the exposure time and concentration were increased. The accumulation study showed the significant increases of both metals when the exposure time and concentration were increased. The roots of S. cucullata had higher Cd and Pb contents than leaves suggesting that the metals were bound to the root cells and were partially transported to the leaves. PMID- 17405326 TI - Detection of current and potential hazelnut plantation areas in Trabzon, North East Turkey using GIS and RS. AB - Monitoring agricultural products requires the periodic determination of land cover and the production of land use policies in an optimum way. The hazelnut is one of the important Turkish agricultural exports and Turkey provides 77% of the world's hazelnuts. In Turkey, hazelnut production exceeds the demand; new regulations have been enacted to create new land use policies. By putting into practice regulations restricting hazelnut plantation areas, a more efficient and productive hazelnut harvest policy could be created. Therefore, more information on existing land cover is required to determine optimum (or ideal) potential hazelnut areas (PHA) and to forecast future crop production. The principle aim of this study is to create a methodology for determining existing PHA, using Geographic information system (GIS) and remote sensing (RS) techniques regarding to support hazelnut policy developers and economists. This study was basically carried out in the province of Trabzon, which is one of the most important hazelnut production areas in Turkey. Landsat ETM+ image was used to generate a current land cover classification. Using the supervised classification method, overall accuracy was determined to be 84.7%. Suitable hazelnut areas were determined according to criteria settled by government regulations. PMID- 17405327 TI - Vinasse biodegradation by Phanerochaete chrysosporium. AB - Vinasse is a colored recalcitrant wastewater of the distillery industry. The aim of this work was to study the use of Phanerochaete chrysosporium for the vinasse degradation under two different growth conditions. Vinasse was treated by P. chrysosporium in a liquid inoculum form, during 32 days at room temperature (approximately 25 degrees C) and at 39 degres C. Chemical oxygen demand (COD), total phenol concentration and color removal were measured and there8 was a decrease in COD, phenolic concentration and color of 47.48%, 54.72% and 45.10% respectively, at room temperature and a decrease in 54.21%, 59.41% and 56.8 1% respectively at 39 degrees C. PMID- 17405328 TI - Effects of cold stratification on germination rate and percentage of caper (Capparis ovata Desf.) seeds. AB - Caper is an important plant because of its high adaptability to marginal agriculture fields that are not suitable for agricultural crops. Different parts of caper such as roots, fruits, flowers and buds can be used to increase the inhabitant's income. The goal of this research is to determine germination rate and percentage of caper according to different duration of cold stratification (10, 20, 30, 40, 50 and 60 days) treatments. Cold stratification procedures under greenhouse condition were applied for eliminating seed dormancy to find the most suitable germination conditions because the presence of seed dormancy causes difficulties in seedling production. The seed germination started and stopped 21 and 57 days after sowing, respectively. While the highest germination percentage (46.6%) was obtained in seeds that were cold stratified for 60 days, the lowest germination percentage (3.6 7%) was determined in control seeds. PMID- 17405329 TI - Effect of cadmium on germination, coleoptile and root growth of barley seeds in the presence of gibberellic acid and kinetin. AB - Effect of cadmium on barley seeds treated with kinetin and gibberellic acid was investigated. As usual, cadmium has inhibited seed germination, and showed important inhibitory effects on roots and coleoptile growth after germination. In general, increase in cadmium concentration caused a greater inhibition of germination, root and coleoptile growth. The adverse effect of cadmium on root and coleoptile growth was more pronounced than that on germination. While testa was pierced by radicle (an indication of germination), no root or coleoptile development was observed above at concentration of 3-9.5 mM CdCl2xH2O. Low concentrations of cadmium have inhibited the root growth more than it did on coleoptile growth. Treatment of seeds with gibberellic acid and kinetin did not show any significant difference on the effect of cadmium in germination. However, inhibition of coleoptile elongation by cadmium has decreased a very much after kinetin application. The same result, although with lower rates when compared to kinetin, has been obtained for GA3 as well. In addition, the inhibitory effect of cadmium on root growth increased even more after kinetin application. The results have been found statistically significant through the least significant different (LSD) test at levels ofp < 0.05 and p < 0.01. PMID- 17405330 TI - Simulation of germination of pioneer species along an experimental drought gradient. AB - The germination of ten plant species from the Iberian Peninsula was assessed along a water deficit gradient between -0. 1652 (moist) and -0.4988 MPa (dry) of osmotic potential, created by addition of increasing concentrations of polyethylene glycol (PEG 6000) to distilled water in which plants were grown hydroponically. The level and rate of germination of Daucus carota and Thapsia villosa significantly decreased with decreasing psi. Seeds of Dactylis glomerata and Dittrichia viscosa had positive germination responses to low osmotic potentials; germination of Epilobium hirsutum was not affected by osmotic potential. Germination of Medicago arabica, Cynosurus cristatus, Cistus ladanifer and Cistus albidus, was no favored by the addition of polyethylene glycol (PEG). Germination of Foeniculum vulgare and Thapsia villosa was inhibited by PEG. PMID- 17405331 TI - Differential response of Tilapia guineensis fingerlings to inorganic fertilizer under various salinity regimes. AB - The influence of salinity on the response of the estuarine teleost, Tilapia guineensis fingerlings to acute toxic effects of inorganic nitrogen-phosphorus potassium (NPK) (15: 15: 15) fertilizer was investigated using semi-static bioassay. The toxicity of NPK fertilizer was found to increase significantly with increase in the salinity level from 0.05 %. to 32.4 %o. The 96 hr LC50 value at salinity of 32.4 %o was 0.11 mg/l and was found to be significantly (p < 0.05) higher than the toxicity values at any other salinity level of media evaluated. The implication of the findings is that pollution control standards and/or safe limits for brackish water ecosystem should consider variations in salinity regimes for greater relevance and reliability. PMID- 17405332 TI - Inverse effects of Polyacrylamide (PAM) usage in furrow irrigation on advance time and deep percolation. AB - The positive effects of Polyacrylamide (PAM), which is used as a soil conditioner in furrow irrigation, on sediment transport, erosion, and infiltration have been investigated intensively in recent years. However, the effects of PAM have not been considered enough in irrigation system planning and design. As a result of increased infiltration because of PAM, advance time may be inversely affected and deep percolation increases. However, advance time in furrow irrigation is a crucial parameter in order to get high application efficiency. In this study, inverse effects of PAM were discussed, and as an alternative solution, the applicability of surge flow was investigated. PAM application significantly increased the advance time at the rates of 41.3-56.3% in the first irrigation. The application of surge flow with PAM removed this negative effect on advance time, where there was no statistically significant difference according to normal continuous flow (without PAM). PAM applications significantly increased the deep percolation, 80.3-117.1%. Surge flow with PAM had significantly positive effect on the deep percolation compared to continuous flow with PAM but not compared to normal continuous flow. These results suggested that irrigation planning should me made based on the new soil and flow conditions because of PAM usage, and surge flow can be a solution to these problems. PMID- 17405333 TI - Groundwater quality of north-east Libya. AB - The quality of groundwater was assessed to their suitability for drinking at six places of north-east Libya viz. El-Marj, Albayda, Shahat, Susa, Ras al-Hilal and Derna, during November, 2003 to March, 2004, by determining their physicochemical parameters (17 parameters) and water quality index (15 parameters). The temperatures of water samples averaged 15.1 degree C, pH values were alkaline and dissolved oxygen values were in safe ranges. Electrical conductivity, total dissolved solids and hardness of water at all places except Susa were within the standard limits. Alkalinity of well water at all six places exceeded, and chlorine and chloride (except Susa) were within the desirable limits set for them. Fluoride and nitrate contaminations of well water were not observed, and only very low values of phosphorus, manganese, chromium, iron and zinc were recorded. Copper in well water was generally high, and at Susa and Ras al-Hilal, it exceeded the desirable limit. Parametric ratios showed that all parameters studied except those of pH, dissolved oxygen, alkalinity and total chlorine originated from sources different from that of hardness. Water quality index (WQI) revealed that well water of Albayda and Shahat were good for drinking and were only slightly polluted, whereas those of El-Marj, Ras al-Hilal and Derna were moderately polluted. However, the well water of Susa was excessively polluted and was unsuitable for drinking. Suitable suggestions were made to improve the quality of groundwater of N.E. Libya. PMID- 17405334 TI - Phytotoxicity of cadmium on the physiological dynamics of Salvinia natans L. grown in macrophyte ponds. AB - The objective of this experiment is to study the potential ability of Salvinia natans L. to use in phytoassay for the cadmium toxicity in ecotoxicological studies. It is a simple and cost competitive method. The sensitivity of S. natans in terms of biochemical changes and enzyme activities to Cd2+ was remarkably noticeable. The catalase and protease activity was significantly decreased in the presence of cadmium, with increase of peroxidase and superoxide dismutase activity. The level of protein, carbohydrate and chlorophyll decreased and proline content increased in plants. The maximum permissible concentration of cadmium in different general water quality standards was evaluated and found that cadmium standard is not stringent enough. PMID- 17405335 TI - Aspergillus niger as a new allergic agent associated with bindis and its efficacy against homeopathic drugs. AB - Aspergillus was found as a dominant fungi to associate with brands of bindis. Among three potencies of four homeopathic drugs, Lycopodium 1M, Sulphur 1M, and Sepia 30 showed maximum inhibition zone of Aspergillus niger in inhibition zone technique. In poison food technique, Sepia 30M, Tellurium 30M, Sulphur 1M and Lycopodium 200 showed maximum percentage inhibition against A. niger PMID- 17405336 TI - Suitability and water quality criteria of an open drainage municipal sewage water at Coimbatore, used for irrigation. AB - Sanganur canal is the major open drainage system which has intricate linkage with storm water supply, domestic sewage and industrial effluent disposal. Water samples from various stations were collected and analysed for physicochemical parameters to assess the water quality of the Sanganur canal system. The study revealed that physicochemical parameters like pH, EC, TDS, DO, BOD, COD exceeded the permissible limit, clearly indicating the need of proper treatment of waste water before discharge into the Noyyal river. PMID- 17405337 TI - Freshwater lakes--a potential source for aquaculture activities--a model study on Perumal Lake, Cuddalore, Tamil Nadu. AB - The freshwater Perumal lake located at Cuddalore was assessed for its suitability and potential for aquaculture practices. Various hydrobiological parameters determined reveals that the various physicochemical characteristics are with in normal range of values. The DO level, BOD and COD values determined in the lake revealed the consequences of community activities and pollution possibilities. The primary productivity data revealed maximum productivity during March which infer that the lake is unaffected by anthropogenic disturbance and community contamination. The bacterial count remained higher during the monsoon periods, which characterize profuse rainfall and storm water discharge into the lake. The microfauna includes zooplankter such as cladocerans, copepods, rotifers and ostracods. Benthos include carps, catfishes, mullets and prawns. The above study revealed that the various parameters in the lake conform to the levels suited for freshwater fish culture and represents a resource for scientific management. PMID- 17405338 TI - Metals and organic components of sewage and sludges. AB - Sewage and sludge samples were collected from rural areas of Vellore district. The metals like Cu, Zn, Cd, Pb, Fe, Ca and Mg were determined by Inductively coupled plasma atomic emission spectrophotometry (ICP-AES). For this determination HCl - HNO3 and 4N HNO3 extracts were used. The total organic carbon (TOC), chemical oxygen demand (COD) and organic matter (OM) have also been determined to evaluate the presence of organic and metallic contents in sewage and sludge environment. PMID- 17405339 TI - Effect of cortisol on female freshwater fish Notopterus notopterus. AB - In the present study, effect of cortisol on the ovary of the freshwater fish Notopterus notopterus was studied during four phases of the reproductive cycle. The hormone was injected 60microg/fish for ten days. Cortisol in fish is known to increase the metabolic activity. After the hormone treatment the ovarian somatic index and the hepatosomatic index increases during non breeding phase. The young oocytes transferred into vitellogenic oocytes in all the phases after cortisol treatment. The cortisol induced increased ovarian activity may be due to increase in the metabolic activity through the involvement of hepatic cells specially during non-breeding period providing support for energy production for reproductive activity in the freshwater fish N. notopterus. PMID- 17405340 TI - Glutathione and associated enzymes in toxic cataractogenesis-selenite model. AB - Glutathione, gamma-glutamyl cysteine synthetase (gamma -GCS) and glutathione reductase (GSH-R) activity were determined biochemically in the lens during various stages after subcutaneous administration of sodium selenite in multiple low dosages and single high dosages. The GSH concentration and gamma-GCS and GSH R activity declined progressively after the selenite administration. The changes observed were discussed in relation to the possible role of selenite interaction with GSH and the enzymes. PMID- 17405341 TI - Lunar rhythm in the planktonic biomass of the Nicco Park lake, Bhubaneswar. AB - The lunar cycle imparts certain stimulatory effects on the rhythmic behaviour of plankton and their life processes by which they actively migrate to the surface and gradually sink as the moon fades. This may also result in morphological changes in organisms, such as Lorica which becomes smaller towards the full moon night by changing the size of postero-lateral spines and surface area. In addition, cyclomorphic forms have been observed in zooplankton, which appears to be a suitable achivement for swarming and for the differentiation of sex by secondary sexual characteristics. Lunar rhythm in plankton appears to be governed by an internal clock, that is the "Biological clock". PMID- 17405342 TI - Deviation of negatively charged protein fractions in the trochophore and veliger larvae by the larvicidal action of baygon in freshwater pulmonate Gyraulus convexiusculus (Planorbidae). AB - In the present investigation egg capsules of Gyraulus convexiusculus were treated with different concentrations of baygon. A dose and duration dependent deviations in the number of negatively charged protein fractions in the trochophore and veliger larval stages were observed. It resulted into anomalies in the morphogenesis and organogenesis of corresponding larval stages. Most of the protein bands showed the decrease in the protein positive intensities in comparison to control. This suggested that baygon causes larval toxicity in Gyraulus convexiusculus. PMID- 17405343 TI - Is simulation based medicine training the future of clinical medicine? AB - The training of physician in the art and science of clinical medicine presents several challenges that are well suited to simulation based medical education (SBME). Modern patient centered medical education seeks to provide comprehensive "hands-on" clinical exposure for physicians in training, while simultaneously providing maximum individual patient comfort and safety. The ethical conundrum is obvious: direct patient contact is needed in order to educate the best clinical physicians and surgeons, but patients have an expectation to be treated and have surgery performed only by highly trained healthcare personnel. This is the kernel of the "medical educators dilemma". Simulation based medical education can partially solve "the medical educators dilemma" by providing realistic medical education in a safe, error tolerant environment with convenience and advantages over conventional "bedside" training but is it real medicine or make believe! PMID- 17405344 TI - Anti-ulceral and anti-oxidative properties of "earthworm paste" of Lampito mauritii (Kinberg) on Rattus Norvegicus. AB - Studies have been made to understand the anti-ulceral and anti-oxidant properties of the "earthworm paste" derived from Lampito mauritii (Kinberg), an indigenous species, in comparison with the standard anti-ulceral drug-ranitidine, on the Wistar strain albino rats Rattus norvegicus. Administration of 200 mg/kg aspirin was found to increase the volume of gastric juice secretion, total acidity, free acidity, ulcer index and reduce the pH. It also had decreased the anti-oxidant levels such as reduced glutathione, glutathione peroxidase, catalase, superoxide dismutase and increased the level of thiobarbituric acid reactive substances. Pretreatment with the standard drug-ranitidine (50 mg/kg) and different doses of "earthworm paste" (20, 40, 80, 160 and 320 mg/kg) in ulcer induced animal had enhanced the pH, decreased the volume of gastric juice, free acidity, total acidity and reduced the ulcer index. Further the activities of reduced glutathione, glutathione peroxidase, catalase, superoxide dismutase were increased whereas the thiobarbituric acid reactive substance had decreased. The results were more significant in rats administered with 160 mg/kg "earthworm paste" than the application of ranitidine and other doses of "earthworm paste". This indicates the presence of antiulcer and anti-oxidative effects in "earthworm paste". In conclusion, administration of 160 mg "earthworm paste"/kg was found to have better therapeutic properties. PMID- 17405345 TI - Modulating effect of Hypericum perforatum extract on astrocytes in MPTP induced Parkinson's disease in mice. AB - The present study has been evaluated the neuroprotective effect of Hypericum perforatum extract on the reaction of astrocytes in mice brain treated with an intraperitoneal injection of 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (20 mg/kg with 2 hr intervals). Treatment with Hypericum perforatum extract (HPE) resulted in an inhibition of monoamine oxidase-B (MAO-B) activity and reduced astrocyte activation in striatal area induced by MPTP. The results show that HPE has neuromodulating effect against MPTP induced Parkinson's disease in mice. PMID- 17405346 TI - Influence of insulin resistance in obese patients on elevated serum alanine aminotransferase. AB - BACKGROUND AND OBJECTIVE: Insulin resistance has been associated with fat liver and non-alcoholic fatty liver disease. The aim of our study was to study the influence of insulin resistance in obese patients on elevated serum alanine aminotransferase. RESEARCH METHODS: A population of 91 obesity non diabetic outpatients was analyzed in a cross sectional study. HOMA-IR was calculated as indicator of insulin resistance. RESULTS: The mean age was 39.2 +/- 16.7 years and the mean BMI 34.4 +/- 5.2. Patients were classified as group I (control, n=74) when serum (Alanine Aminotransferase) ALT activity was normal or group II (NAFLD, n=17) when serum ALT activity was greater than the upper limit of normal reference laboratory (> or =43 UI/L). Waist circumference, fat mass and hip to waist ratio were higher in group II. Insulin (13.5 +/- 7.8 mUI/L vs 24.9 +/- 16.7 mUI/L; p < 0.05), HOMA-IR (1.9 +/- 1.1 vs 3.9 +/- 2.8), and triglycerides levels (115.1 +/- 66.8 mg/dl vs. 153.2 +/- 71.2 mg/dl; p < 0.05) were higher in group II than group I. In the logistic regression analysis with a dependent dicotomic variable (ALT; group I and group II), the HOMA-IR remained in the model, with an Odd's Ratio to develop ALT >43 U/L of 2.18 (CI:95%: 1.12-4.2) with each 1 unit of HOMA-IR adjusted by age, sex, weight, and dietary intake. CONCLUSION: Insulin resistance in obese patients is associated with ALT activity. Further study is needed to evaluate histological changes and new treatments in these patients. PMID- 17405347 TI - Oxidative stress detection: what for? Part II. AB - Oxygen-free radicals, more generally known as reactive oxygen species (ROS) along with nitrogen species (RNS) are well recognised for playing a dual role both deleterious and beneficial species. The cumulative generation of ROS/RNS through either endogenous or exogenous insults is termed oxidative stress and is common for many types of diseases that are linked with altered redox regulation of cellular signalling pathways. PMID- 17405348 TI - Nasopharyngeal adenoid cystic carcinoma: a rare nasopharyngeal tumor. AB - Adenoid cystic carcinoma (ACC) is the most common histologic subtype of the salivary gland malignancies, but it is very rare in the nasopharynx with relatively few cases reported in the literature. We report a case of a 57 year old woman with an ACC in the right Rosenmuller fossa of the nasopharynx, treated with combined radiotherapy and chemotherapy, without any recurrence or intracranial spread detected until the recent follow-up. PMID- 17405349 TI - Apoptosis induction by quinazoline-derived alpha1-blockers in prostate cancer cells: biomolecular implications and clinical relevance. AB - In patients with androgeri-sensitive prostate cancer androgen-deprivation therapy leads to apoptosis of hormone-dependent cells but it may select out androgen refractory clones. The apoptosis of prostate cancer cells induced by quinazoline derived alpha1-adrenoceptor antagonists (doxazosin, terazosin, prazosin) counteracts cell proliferation and may have the potential of reversing or delaying prostate cancer growth. Elucidation of the biomolecular mechanisms whereby quinazoline-alpha1-adrenoceptor-antagonists are able to induce anoikis and apoptosis in androgen-refractory prostate cancer cells would be favorable for designing effective therapeutic regimens in the management of advanced prostate cancer. PMID- 17405350 TI - Loperamide cause of prolonged urinary retention after acute gastroenteritis. AB - Urinary retention is uncommon in children and only one case has been described in literature with loperamide treatment. We report the occurrence of prolonged urinary retention in a 10-years-old girl after receiving oral loperamide for an acute gastroenteritis. The first episode of urinary retention lasted for 24 hours; radiological evaluation (Magnetic Resonance, cistography and renal ultrasound) did not find abnormality; it was necessary deplete bladder with catheterism. Previous to the gastroenteritis, diuresis has always been regular and she did not suffer for any neurological or urinary problems. Patient was submitted to, without success, percutaneous posterior tibial nerve stimulation (SANS) and recovered after one year clean intermittent catheterization. CONCLUSION: In a period of increasing popularity of self-medical therapy for common children's diseases, paediatricians should be aware of the potential, rare and chronic effects of this type of treatment. Moreover our case suggests that reaction to loperamide should be added to the etiological list of prolonged urinary retention in young patients. PMID- 17405351 TI - Anaplastic thyroid carcinoma: a case of a young women that reports a survival exceeding 6 years. AB - Anaplastic thyroid carcinoma (ATC) is an uncommun and highly aggressive malignancy. Differentiated follicular and papillary thyroid carcinomas account for most (80-90%) thyroid malignancies with ATC accounting for less than 5%. A diagnosis of ATC is usually fatal with a mean survival of 3-9 months and only 10 15% alive at 2 years. Histologic examination reveals that many of them contain a papillary structure or follicular components in focal areas and genetic alteration is the driving for genesis of cancer and progression. These studies showing that ATC represents a terminal "de-differentiation" of pre-existing differentiated carcinoma. Most of patients are incurable, nevertheless a multimodality approach, incorporating surgery with the aim to obtain complete macroscopic resection and achieve clear resections margins followed by adjuvant treatment RT (radiotherapy) and/or chemotherapy, improve local control and extend the survival. The aim of the present study has been to review a case of a young women with ATC that reports a survival exceeding 6 years. PMID- 17405352 TI - A comparative epidemiological study of hantavirus infection in Japan and Far East Russia. AB - Hantaviruses are causative agents of some severe human illnesses, including hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). The viruses are maintained by rodent hosts, and humans acquire infection by inhaling virus-contaminated excreta from infected animals. To examine the epidemiology of hantavirus infections in Japan and Far East Russia, we conducted epidemiological surveys in these regions. In Japan, anti-hantavirus antibodies were found in four rodent species, Clethrionomys rufocanus, Rattus norvegicus, R. rattus, and Apodemus speciosus. Although no new HFRS cases have been officially reported over the past 20 years in Japan, one member of the Japan Ground Self Defense Force did test positive for hantavirus antibody. Repeated surveys in Far East Russia have revealed that two distinct hantavirus types cause severe HFRS in this region. Hantavirus sequences identified from A. peninsulae, fetal HFRS cases in Vladivostok, and Amur virus are highly similar to each other (> 92% identity), but they are less similar (approximately 84% identity) to the prototypical Hantaan virus, which is carried by A. agrarius. Phylogenetic analysis also indicates that Amur and A. peninsulae-associated viruses are distinct from Hantaan virus, suggesting that A. peninsulae is the reservoir animal for Amur virus, which causes severe HFRS. From HFRS patients in the Khabarovsk region, we identified viruses with nucleotide sequences that are more similar to Far East virus (> 96%identity) than to the Hantaan (88-89% identity) or Amur (81-83% identity) viruses. Phylogenetic analysis also indicates that the viruses from Khabarovsk HFRS patients are closely related to the Far East virus, and distinct from Amur virus. PMID- 17405353 TI - Oxidative stress and bovine liver diseases: role of glutathione peroxidase and glucose 6-phosphate dehydrogenase. AB - This article summarizes the different types of free radicals, antioxidants and the effect of oxidative stress on the activities of glutathione peroxidase and glucose 6-phosphate dehydrogenase in bovine liver diseases. A growing body of evidence suggests that the formation of reactive oxygen species is a common occurrence associated with most if not all disease processes. The overall importance of reactive oxygen species to the progression and severity of various disease states varies greatly depending on the conditions and whether the disease is acute or chronic. Free radical researches in animals are in progress and further investigations are needed to establish the involvement of reactive oxygen species in diseases affecting different animal species and the pathology they produce. PMID- 17405354 TI - Epizootiological survey of Trichinella spp. infection in carnivores, rodents and insectivores in Hokkaido, Japan. AB - In order to evaluate the present epidemiological situation of Trichinella infection in wild animals in Hokkaido, Japan, red foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) , brown bears (Ursus arctos) , martens (Martes melampus), rodents and insectivores captured in Hokkaido were examined for muscle larvae by the artificial digestion method from 2000 to 2006. Foxes (44/319, 13.8%), raccoon dogs (6/77, 7.8%) and brown bears (4/126, 3.2%) were found to be infected with Trichinella larvae and all other animal species evaluated were negative. Multiplex PCR and DNA sequencing revealed that larvae from a fox captured in Otofuke, in south-eastern Hokkaido, were T. nativa, and larvae from 27 animals including 21 foxes, 2 raccoon dogs and 4 brown bears captured in western Hokkaido were Trichinella T9. PMID- 17405355 TI - Leptin inhibits hepatocyte growth factor-induced ductal morphogenesis of bovine mammary epithelial cells. AB - We examined the effect of stroma-derived factors, hepatocyte growth factor (HGF) and leptin, on morphological differentiation of bovine mammary epithelial cells (BMEC) in collagen gel three-dimensional culture in vitro. BMEC treated with HGF, but not leptin, formed duct-like organoids. The formation of organoids by HGF was enhanced by treatment with a mixture of insulin, cortisol and prolactin, while BMEC treated with the mixture alone did not produce the organoid. In contrast, the formation of organoids by HGF was dose-dependently inhibited by simultaneous addition of leptin, regardless of the presence or absence of the hormone mixture. These results suggest that stroma-derived factors intricately regulate mammary epithelial morphogenesis. PMID- 17405356 TI - Lumenal localization in the endoplasmic reticulum of the C-terminal tail of an AE1 mutant responsible for hereditary spherocytosis in cattle. AB - An R664X nonsense mutant AE1 is responsible for dominant hereditary spherocytosis in cattle and is degraded by the proteasomal endoplasmic reticulum-associated degradation. The present study demonstrated that R664X AE1 translated in vitro had the trypsin-sensitve site identical to that of the wild-type AE1. The P661S/R664X mutant containing a possible N-glycosylation site at Asn660 showed an increase in size by 3 kDa both in the cell-free translation system and in transfected HEK293 cells. Moreover, steady state levels of R664X and P661S/R664X in HEK293 cells were markedly increased in the presence of a proteasome inhibitior. These findings indicate that the truncated C-terminal region of R664X AE1 has lumenal localization in the endoplasmic reticulum and is not accessible to proteasomal machineries in the cytosol. PMID- 17405359 TI - Successive therapies may extend survival in AIPC. Interview by Alice Goodman. PMID- 17405360 TI - Aortic dissection. PMID- 17405361 TI - Finding the words. How to handle difficult conversations. PMID- 17405362 TI - Spores, babies, and alcohol. A nurse's battle with C. diff. PMID- 17405363 TI - "Driver's ed" for the OR nurse. PMID- 17405364 TI - "Please don't let me die!" One patient's love of life inspired me. PMID- 17405365 TI - Mathematical model for the hemodynamic response to venous occlusion measured with near-infrared spectroscopy in the human forearm. AB - We propose a mathematical model to describe the hemodynamic changes induced by a venous occlusion in a human limb. These hemodynamic changes, which include an increase in blood volume, a reduction in blood flow, and modifications to the oxygen saturation of hemoglobin, can all be measured noninvasively with near infrared spectroscopy (NIRS). To test the model, we have performed NIRS measurements on the human forearm, specifically on the brachioradialis muscle, during venous occlusion induced by a pneumatic cuff inflated around the upper arm to pressures within the range 10-60 mmHg. We have found a good agreement between parameters measured by NIRS (total hemoglobin concentration and hemoglobin saturation) and the corresponding model parameters (capacitor voltage and arterial/capillary branch current). In particular, model and experiment indicate that the time constant for blood accumulation during venous occlusion (approximately 73-79 s) is much slower than the time constant for blood drainage following cuff release (approximately 5 s). These results indicate that this mathematical model can be a valuable analytical tool to characterize, optimize, and further develop diagnostic measurement schemes that use venous occlusion approaches. PMID- 17405366 TI - Algebraic multigrid preconditioner for the cardiac bidomain model. AB - The bidomain equations are considered to be one of the most complete descriptions of the electrical activity in cardiac tissue, but large scale simulations, as resulting from discretization of an entire heart, remain a computational challenge due to the elliptic portion of the problem, the part associated with solving the extracellular potential. In such cases, the use of iterative solvers and parallel computing environments are mandatory to make parameter studies feasible. The preconditioned conjugate gradient (PCG) method is a standard choice for this problem. Although robust, its efficiency greatly depends on the choice of preconditioner. On structured grids, it has been demonstrated that a geometric multigrid preconditioner performs significantly better than an incomplete LU (ILU) preconditioner. However, unstructured grids are often preferred to better represent organ boundaries and allow for coarser discretization in the bath far from cardiac surfaces. Under these circumstances, algebraic multigrid (AMG) methods are advantageous since they compute coarser levels directly from the system matrix itself, thus avoiding the complexity of explicitly generating coarser, geometric grids. In this paper, the performance of an AMG preconditioner (BoomerAMG) is compared with that of the standard ILU preconditioner and a direct solver. BoomerAMG is used in two different ways, as a preconditioner and as a standalone solver. Two 3-D simulation examples modeling the induction of arrhythmias in rabbit ventricles were used to measure performance in both sequential and parallel simulations. It is shown that the AMG preconditioner is very well suited for the solution of the bidomain equation, being clearly superior to ILU preconditioning in all regards, with speedups by factors in the range 5.9-7.7. PMID- 17405367 TI - Estimation of hidden state variables of the Intracranial system using constrained nonlinear Kalman filters. AB - Impeded by the rigid skull, assessment of physiological variables of the intracranial system is difficult. A hidden state estimation approach is used in the present work to facilitate the estimation of unobserved variables from available clinical measurements including intracranial pressure (ICP) and cerebral blood flow velocity (CBFV). The estimation algorithm is based on a modified nonlinear intracranial mathematical model, whose parameters are first identified in an offline stage using a nonlinear optimization paradigm. Following the offline stage, an online filtering process is performed using a nonlinear Kalman filter (KF)-like state estimator that is equipped with a new way of deriving the Kalman gain satisfying the physiological constraints on the state variables. The proposed method is then validated by comparing different state estimation methods and input/output (I/O) configurations using simulated data. It is also applied to a set of CBFV, ICP and arterial blood pressure (ABP) signal segments from brain injury patients. The results indicated that the proposed constrained nonlinear KF achieved the best performance among the evaluated state estimators and that the state estimator combined with the I/O configuration that has ICP as the measured output can potentially be used to estimate CBFV continuously. Finally, the state estimator combined with the I/O configuration that has both ICP and CBFV as outputs can potentially estimate the lumped cerebral arterial radii, which are not measurable in a typical clinical environment. PMID- 17405368 TI - Hybrid finite element method for describing the electrical response of biological cells to applied fields. AB - A novel hybrid finite element method (FEM) for modeling the response of passive and active biological membranes to external stimuli is presented. The method is based on the differential equations that describe the conservation of electric flux and membrane currents. By introducing the electric flux through the cell membrane as an additional variable, the algorithm decouples the linear partial differential equation part from the nonlinear ordinary differential equation part that defines the membrane dynamics of interest. This conveniently results in two subproblems: a linear interface problem and a nonlinear initial value problem. The linear interface problem is solved with a hybrid FEM. The initial value problem is integrated by a standard ordinary differential equation solver such as the Euler and Runge-Kutta methods. During time integration, these two subproblems are solved alternatively. The algorithm can be used to model the interaction of stimuli with multiple cells of almost arbitrary geometries and complex ion channel gating at the plasma membrane. Numerical experiments are presented demonstrating the uses of the method for modeling field stimulation and action potential propagation. PMID- 17405369 TI - Acoustic analysis and detection of hypernasality using a group delay function. AB - In this paper, we describe a group delay-based signal processing technique for the analysis and detection of hypernasal speech. Our preliminary acoustic analysis on nasalized vowels shows that, even though additional resonances are introduced at various frequency locations, the introduction of a new resonance in the low-frequency region (around 250 Hz) is found to be consistent. This observation is further confirmed by a perceptual analysis carried out on vowel sounds that are modified by introducing different nasal resonances, and an acoustic analysis on hypernasal speech. Based on this, for subsequent experiments the focus is given only to the low-frequency region. The additive property of the group delay function can be exploited to resolve two closely spaced formants. However, when the formants are very close with considerably wider bandwidths as in hypernasal speech, the group delay function also fails to resolve. To overcome this, we suggest a band-limited approach to estimate the locations of the formants. Using the band-limited group delay spectrum, we define a new acoustic measure for the detection of hypernasality. Experiments are carried out on the phonemes /a/, /i/, and /u/ uttered by 33 hypernasal speakers and 30 normal speakers. Using the group delay-based acoustic measure, the performance on a hypernasality detection task is found to be 100% for /a/, 88.78% for /i/ and 86.66% for /u/. The effectiveness of this acoustic measure is further cross verified on a speech data collected in an entirely different recording environment. PMID- 17405370 TI - Noise cancellation signal processing method and computer system for improved real time electrocardiogram artifact correction during MRI data acquisition. AB - A system was developed for real-time electrocardiogram (ECG) analysis and artifact correction during magnetic resonance (MR) scanning, to improve patient monitoring and triggering of MR data acquisitions. Based on the assumption that artifact production by magnetic field gradient switching represents a linear time invariant process, a noise cancellation (NC) method is applied to ECG artifact linear prediction. This linear prediction is performed using a digital finite impulse response (FIR) matrix, that is computed employing ECG and gradient waveforms recorded during a training scan. The FIR filters are used during further scanning to predict artifacts by convolution of the gradient waveforms. Subtracting the artifacts from the raw ECG signal produces the correction with minimal delay. Validation of the system was performed both off-line, using prerecorded signals, and under actual examination conditions. The method is implemented using a specially designed Signal Analyzer and Event Controller (SAEC) computer and electronics. Real-time operation was demonstrated at 1 kHz with a delay of only 1 ms introduced by the processing. The system opens the possibility of automatic monitoring algorithms for electrophysiological signals in the MR environment. PMID- 17405371 TI - ECG beat detection using a geometrical matching approach. AB - In the framework of the electrocardiography (ECG) signals, this paper describes an original approach to identify heartbeat morphologies and to detect R-wave events. The proposed approach is based on a "geometrical matching" rule evaluated using a decision function in a local moving-window procedure. The decision function is a normalized measurement of a similarity criterion comparing the windowed input signal with the reference beat-pattern into a nonlinear-curve space. A polynomial expansion model describes the reference pattern. For the curve space, an algebraic-fitting distance is built according to the canonical equation of the unit circle. The geometrical matching approach operates in two stages, i.e., training and detection ones. In the first stage, a learning-method based on genetic algorithms allows us estimating the decision function from training beat-pattern. In the second stage, a level-detection algorithm evaluates the decision function to establish the threshold of similarity between the reference pattern and the input signal. Finally, the findings for the MIT-BIH Arrhythmia Database present about 98% of sensitivity and 99% of positive predictivity for the R-waves detection, using low-order polynomial models. PMID- 17405372 TI - A framework for the analysis of acoustical cardiac signals. AB - Skilled cardiologists perform cardiac auscultation, acquiring and interpreting heart sounds, by implicitly carrying out a sequence of steps. These include discarding clinically irrelevant beats, selectively tuning in to particular frequencies and aggregating information across time to make a diagnosis. In this paper, we formalize a series of analytical stages for processing heart sounds, propose algorithms to enable computers to approximate these steps, and investigate the effectiveness of each step in extracting relevant information from actual patient data. Through such reasoning, we provide insight into the relative difficulty of the various tasks involved in the accurate interpretation of heart sounds. We also evaluate the contribution of each analytical stage in the overall assessment of patients. We expect our framework and associated software to be useful to educators wanting to teach cardiac auscultation, and to primary care physicians, who can benefit from presentation tools for computer assisted diagnosis of cardiac disorders. Researchers may also employ the comprehensive processing provided by our framework to develop more powerful, fully automated auscultation applications. PMID- 17405373 TI - P-wave morphology assessment by a gaussian functions-based model in atrial fibrillation patients. AB - Aim of this study was to present a P-wave model, based on a linear combination of Gaussian functions, to quantify morphological aspects of P-wave in patients prone to atrial fibrillation (AF). Five-minute ECG recordings were performed in 25 patients with permanent dual chamber pacemakers. Patients were divided into high risk and low-risk groups, including patients with and without AF episodes in the last 6 mo preceding the study, respectively. ECG signals were acquired using a 32 lead mapping system for high-resolution biopotential measurement (ActiveTwo, Biosemi, The Netherlands, sample frequency 2 kHz, 24-bit resolution). Up to 8 Gaussian models have been computed for each averaged P-wave extracted from every lead. The P-wave morphology was evaluated by extracting seven parameters. Classical time-domain parameters, based on P-wave duration estimation, have been also estimated. We found that the P-wave morphology can be effectively modeled by a linear combination of Gaussian functions. In addition, the combination of time domain and morphological parameters extracted from the Gaussian function-based model of the P-wave improves the identification of patients having different risks of developing AF. PMID- 17405374 TI - Electrocardiogram based neonatal seizure detection. AB - A method for the detection of seizures in the newborn using the electrocardiogram (ECG) signal is presented. Using a database of eight recordings, a method was developed for automatically annotating each 1-min epoch as "nonseizure" or "seizure". The system uses a linear discriminant classifier to process 41 heartbeat timing interval features. Performance assessment of the method showed that on a patient-specific basis an average accuracy of 70.5% was achieved in detecting seizures with associated sensitivity of 62.2% and specificity of 71.8%. On a patient-independent basis the average accuracy was 68.3% with sensitivity of 54.6% and specificity of 77.3%. Shifting the decision threshold for the patient independent classifier allowed an increase in sensitivity to 78.4% at the expense of decreased specificity (51.6%), leading to increased false detections. The results of our ECG-based method are comparable with those reported for EEG-based neonatal seizure detection systems and offer the benefit of an easier acquisition methodology for seizure detection. PMID- 17405375 TI - Estimation of elbow-induced wrist force with EMG signals using fast orthogonal search. AB - In many studies and applications that include direct human involvement-such as human-robot interaction, control of prosthetic arms, and human factor studies hand force is needed for monitoring or control purposes. The use of inexpensive and easily portable active electromyogram (EMG) electrodes and position sensors would be advantageous in these applications compared to the use of force sensors, which are often very expensive and require bulky frames. Multilayer perceptron artificial neural networks (MLPANN) have been used commonly in the literature to model the relationship between surface EMG signals and muscle or limb forces for different anatomies. This paper investigates the use of fast orthogonal search (FOS), a time-domain method for rapid nonlinear system identification, for elbow induced wrist force estimation. It further compares the forces estimated using FOS with the forces estimated by MLPANN for the same human anatomy under an ensemble of operational conditions. In this paper, the EMG signal readings from upper arm muscles involved in elbow joint movement and sensed elbow angular position and velocity are utilized as inputs. A single degree-of-freedom robotic experimental testbed has been constructed and used for data collection, training and validation. PMID- 17405376 TI - Myoelectric signal classification for phoneme-based speech recognition. AB - Traditional acoustic speech recognition accuracies have been shown to deteriorate in highly noisy environments. A secondary information source is exploited using surface myoelectric signals (MES) collected from facial articulatory muscles during speech. Words are classified at the phoneme level using a hidden Markov model (HMM) classifier. Acoustic and MES data was collected while the words "zero" through "nine" were spoken. An acoustic expert classified the 18 formative phonemes in low noise levels [signal-to-noise ratio (SNR) of 17.5 dB] with an accuracy of 99%, but deteriorated to approximately 38% under simulations with SNR approaching 0 dB. A fused acoustic-myoelectric multiexpert system, without knowledge of SNR, improved on acoustic classification results at all noise levels. A multiexpert system, incorporating SNR information, obtained accuracies of 99% at low noise levels while maintaining accuracies above 94% during low SNR (0 dB) simulations. Results improve on previous full word MES speech recognition accuracies by almost 10%. PMID- 17405377 TI - A reconstruction algorithm for breast cancer imaging with electrical impedance tomography in mammography geometry. AB - The conductivity and permittivity of breast tumors are known to differ significantly from those of normal breast tissues, and electrical impedance tomography (EIT) is being studied as a modality for breast cancer imaging to exploit these differences. At present, X-ray mammography is the primary standard imaging modality used for breast cancer screening in clinical practice, so it is desirable to study EIT in the geometry of mammography. This paper presents a forward model of a simplified mammography geometry and a reconstruction algorithm for breast tumor imaging using EIT techniques. The mammography geometry is modeled as a rectangular box with electrode arrays on the top and bottom planes. A forward model for the electrical impedance imaging problem is derived for a homogeneous conductivity distribution and is validated by experiment using a phantom tank. A reconstruction algorithm for breast tumor imaging based on a linearization approach and the proposed forward model is presented. It is found that the proposed reconstruction algorithm performs well in the phantom experiment, and that the locations of a 5-mm-cube metal target and a 6-mm-cube agar target could be recovered at a target depth of 15 mm using a 32 electrode system. PMID- 17405378 TI - Guiding the surgical gesture using an electro-tactile stimulus array on the tongue: a feasibility study. AB - Under conventional "open-" surgery, the physician has to take care of the patient, interact with other clinicians and check several monitoring devices. Nowadays, the computer assisted surgery proposes to integrate 3-D cameras in the operating theatre in order to assist the surgeon in performing minimally invasive surgical punctures. The cameras localize the needle and the computer guides the surgeon towards an intracorporeal clinically defined target. A visualization system (screen) is employed to provide the surgeon with indirect visual spatial information about the intracorporeal positions of the needle. The present work proposes to use another sensory modality to guide the surgeon, thus keeping the visual modality fully dedicated to the surgical gesture. For this, the sensory substitution paradigm using the Bach-y-Rita's "Tongue Display Unit" (TDU) is exploited to provide to the surgeon information of the position tool. The TDU device is composed of a 6 x 6 matrix of electrodes transmitting electrotactile information on the tongue surface. The underlying idea consists in transmitting information about the deviation of the needle movement with regard to a preplanned "optimal" trajectory. We present an experiment assessing the guidance effectiveness of an intracorporeal puncture under TDU guidance with respect to the performance evidenced under a usual visual guidance system. PMID- 17405379 TI - ECG recording on a bed during sleep without direct skin-contact. AB - A new indirect contact (IDC) electrocardiogram (ECG) measurement method (IDC-ECG) for monitoring ECG during sleep that is adequate for long-term use is provided. The provided method did not require any direct conductive contact between the instrument and bare skin. This method utilizes an array of high-input-impedance active electrodes fixed on the mattress and an indirect-skin-contact ground made of a large conductive textile sheet. A thin cotton bedcover covered the mattress, electrodes, and conductive textile, and the participants were positioned on the mattress over the bedcover. An ECG was successfully obtained, although the signal quality was lower and the motion artifact was larger than in conventional direct contact measurements (DC-ECG). The results showed that further studies are required to apply the provided method to an ECG diagnosis of cardiovascular diseases. However, currently the method can be used for HRV assessment with easy discrimination of R-peaks. PMID- 17405380 TI - Safety of active implantable devices during MRI examinations: a finite element analysis of an implantable pump. AB - The goal of this study was to propose a general numerical analysis methodology to evaluate the magnetic resonance imaging (MRI)-safety of active implants. Numerical models based on the finite element (FE) technique were used to estimate if the normal operation of an active device was altered during MRI imaging. An active implanted pump was chosen to illustrate the method. A set of controlled experiments were proposed and performed to validate the numerical model. The calculated induced voltages in the important electronic components of the device showed dependence with the MRI field strength. For the MRI radiofrequency fields, significant induced voltages of up to 20 V were calculated for a 0.3T field strength MRI. For the 1.5 and 3.0OT MRIs, the calculated voltages were insignificant. On the other hand, induced voltages up to 11 V were calculated in the critical electronic components for the 3.0T MRI due to the gradient fields. Values obtained in this work reflect to the worst case situation which is virtually impossible to achieve in normal scanning situations. Since the calculated voltages may be removed by appropriate protection circuits, no critical problems affecting the normal operation of the pump were identified. This study showed that the proposed methodology helps the identification of the possible incompatibilities between active implants and MR imaging, and can be used to aid the design of critical electronic systems to ensure MRI-safety. PMID- 17405381 TI - A 3-D hp finite/infinite element method to calculate power deposition in the human head. AB - The electromagnetic power deposition and transfer properties of a G1 continuous head model reconstructed from magnetic resonance imaging (MRI) data are investigated by using the coupled hp finite/infinite element (FE/IE) method. The discretization error is controlled by a self-adaptive process driven by an explicit a posteriori error estimate. Based on the benchmark problem of reproducing the Mie series solution, the scattering of a plane wave on the curvilinear head model is used to evaluate the hp FE/IE approach and calibrate the error bound. The radiation pattern from a short dipole antenna modeling a cell phone, is analyzed in terms of the level and distribution of the specific absorption rates (SAR). The numerical experiments show that the hybrid hp FE/IE implementation is a competitive tool for accurate assessment of human electromagnetic exposure. PMID- 17405382 TI - Multiple channel detection of steady-state visual evoked potentials for brain computer interfaces. AB - In this paper, novel methods for detecting steady-state visual evoked potentials using multiple electroencephalogram (EEG) signals are presented. The methods are tailored for brain-computer interfacing, where fast and accurate detection is of vital importance for achieving high information transfer rates. High detection accuracy using short time segments is obtained by finding combinations of electrode signals that cancel strong interference signals in the EEG data. Data from a test group consisting of 10 subjects are used to evaluate the new methods and to compare them to standard techniques. Using 1-s signal segments, six different visual stimulation frequencies could be discriminated with an average classification accuracy of 84%. An additional advantage of the presented methodology is that it is fully online, i.e., no calibration data for noise estimation, feature extraction, or electrode selection is needed. PMID- 17405383 TI - Independent component analysis of high-density electromyography in muscle force estimation. AB - Accurate force prediction from surface electromyography (EMG) forms an important methodological challenge in biomechanics and kinesiology. In a previous study (Staudenmann et al., 2006), we illustrated force estimates based on analyses lent from multivariate statistics. In particular, we showed the advantages of principal component analysis (PCA) on monopolar high-density EMG (HD-EMG) over conventional electrode configurations. In the present study, we further improve force estimates by exploiting the correlation structure of the HD-EMG via independent component analysis (ICA). HD-EMG from the triceps brachii muscle and the extension force of the elbow were measured in 11 subjects. The root mean square difference (RMSD) and correlation coefficients between predicted and measured force were determined. Relative to using the monopolar EMG data, PCA yielded a 40% reduction in RMSD. ICA yielded a significant further reduction of up to 13% RMSD. Since ICA improved the PCA-based estimates, the independent structure of EMG signals appears to contain relevant additional information for the prediction of muscle force from surface HD-EMG. PMID- 17405384 TI - Dynamic model of communicating hydrocephalus for surgery simulation. AB - We propose a dynamic model of cerebrospinal fluid and intracranial pressure regulation. In this model, we investigate the coupling of biological parameters with a 3-D model, to improve the behavior of the brain in surgical simulators. The model was assessed by comparing the simulated ventricular enlargement with a patient case study of communicating hydrocephalus. In our model, cerebro-spinal fluid production-resorption system is coupled with a 3-D representation of the brain parenchyma. We introduce a new bi-phasic model of the brain (brain tissue and extracellular fluid) allowing for fluid exchange between the brain extracellular space and the venous system. The time evolution of ventricular pressure has been recorded on a symptomatic patient after closing the ventricular shunt. A finite element model has been built based on a computed tomography scan of this patient, and quantitative comparisons between experimental measures and simulated data are proposed. PMID- 17405385 TI - Capacitive sensing of electrocardiographic potential through cloth from the dorsal surface of the body in a supine position: a preliminary study. AB - A method for obtaining electrocardiographic potential through thin cloth inserted between the measuring electrodes and the skin of a subject's dorsal surface when lying supine has been proposed. The method is based on capacitive coupling involving the electrode, the cloth, and the skin. Examination of a pilot device which employed the method revealed the following: (1) In spite of the gain attenuation in the high frequency region, the proposed method was considered useful for monitoring electrogardiogram (ECG) for nondiagnostic purpose. (2) The method was able to yield a stable ECG from a subject at rest for at least 7 h, and there was no significant adverse effect of long-term measurement on the quality of the signal obtained. (3) Electrode area was the factor that had most influence on the signal, compared with other factors such as cloth thickness and coupling pressure, but could be reduced to 10 cm2 for heart rate detection. (4) Input capacitance of the device was assumed to be the dominant factor for the gain attenuation in the high frequency region, and should be reduced with a view to diagnostic use. Although there is still room for improvement in terms of practical use, the proposed method appears promising for application to bedding as a noninvasive and awareness-free method for ECG monitoring. PMID- 17405386 TI - Wavelet packets feasibility study for the design of an ECG compressor. AB - Most of the recent electrocardiogram (ECG) compression approaches developed with the wavelet transform are implemented using the discrete wavelet transform. Conversely, wavelet packets (WP) are not extensively used, although they are an adaptive decomposition for representing signals. In this paper, we present a thresholding-based method to encode ECG signals using WP. The design of the compressor has been carried out according to two main goals: (1) The scheme should be simple to allow real-time implementation; (2) quality, i.e., the reconstructed signal should be as similar as possible to the original signal. The proposed scheme is versatile as far as neither QRS detection nor a priori signal information is required. As such, it can thus be applied to any ECG. Results show that WP perform efficiently and can now be considered as an alternative in ECG compression applications. PMID- 17405387 TI - Price transparency: building community trust. AB - With the push from policymakers, payers, and consumers for hospitals to make their prices public, healthcare executives need to recognize two central issues related to price transparency: 1) meaningful price transparency involves helping patients and consumers understand their financial obligation for an episode of care, and 2) price transparency is key to the most critical success strategy for healthcare providers: building trust. This article reviews the history of pricing and billing practices and explores why price transparency is not easily achieved in today's environment. Pricing is a mystery even to those of us who work in the field, yet despite its complexity, the call for price transparency is not going to go away. For transparency, the goal should be to establish a rational pricing system that is easily explainable and justified to all stakeholders. Healthcare executives must make pricing a priority, understand cost, develop a pricing philosophy, understand the overall revenue requirements, examine market conditions and prices, and set up systems for review. A rational process of price setting should enhance community trust. In this matter there is nothing less at stake than the hearts of our community members. PMID- 17405388 TI - Transparency--"Deal or no deal"? AB - In the United States, transparency is becoming an ideal worthy of Mom and apple pie, like quality in healthcare. Physicians, payers, hospitals, business associations, and organizations representing patients have all chimed in expressing support. At the local, state, and national levels a variety of transparency initiatives are under way. How will transparency affect the healthcare industry? Transparency could profoundly change today's balance of power, for it is about information, and information is power. As employers push more cost sharing to workers, hospitals and health systems will have to construct a pricing structure that is meaningful to consumers. What are providers to do? To be successful with this new demand, providers should make sure they are making quality information as well as pricing information available to consumers. They will have to know the market, know what their own prices mean, consider the customer, and reengineer business processes around the patient rather than around the billing side of business. PMID- 17405389 TI - Price transparency begins at home. PMID- 17405390 TI - Price transparency's role in healthcare purchasing reform. PMID- 17405391 TI - Transparency--will it lead us back to the future? PMID- 17405392 TI - Nurses' perceptions of the impact of a renal nursing professional practice model on nursing outcomes, characteristics of practice environments and empowerment- Part I. AB - A transformational nursing professional practice model (PPM) was developed and implemented in the London Health Sciences Centre's renal program. The purpose of this study was to examine the impact of a renal nursing PPM on nurses'perceptions of empowerment, characteristics of practice environments, and nursing outcomes. Quantitative and qualitative methodologies were used in this study. This paper will focus primarily on the quantitative results. The qualitative results are presented in Part II (Harwood, Ridley, Lawrence-Murphy, White, Laschinger, Bevan, & O'Brien, in press). A "then-and-now" design was used. There was a significant (p = .005) improvement post-PPM implementation in the nursing foundations for quality of care subscale of the Nursing Worklife Index-Practice Environment Scale (NWI-PES) and organizational relationships (p = .016) measured by the Conditions of Work Effectiveness II (CWEQ-II) Questionnaires. This study provides evidence for PPMs and primary nursing as effective frameworks to positively impact nursing and patient outcomes in a hemodialysis unit. PMID- 17405393 TI - Know how it works before you fix it: a data analysis strategy from an inpatient nephrology patient-flow improvement project. AB - This article describes a two-part data analysis strategy that was used as part of a process improvement project on an inpatient renal unit. The goal of the project was to improve patient flow-through from admission to discharge. The Canadian Institute for Health Information reported that the proportion of incident end stage renal disease in the 65+ age group was on the rise, and that the fastest growing segment of incident dialysis patients was in the 75+ age group. Health professionals who provide hospital services to nephrology patients should be alert to this information and anticipate longer mean lengths of hospital stay and more frequent discharge delays. There has been no research in this area specific to inpatient renal units. The author shares his data analysis strategy in hopes that it will spur more research and other process improvement projects. PMID- 17405394 TI - Counseling about vitamin D supplementation for infants: results of a survey of pediatric and family medicine physicians in Connecticut. AB - BACKGROUND: Vitamin D is essential to the health of children. The American Academy of Pediatrics (AAP) has recently updated its recommendations for vitamin D supplementation. The current level of vitamin D counseling for infants by family medicine physicians and pediatricians in Connecticut is unknown. METHODS: Some 841 self-administered surveys were sent to pediatric primary care providers in Connecticut to evaluate the frequency of counseling regarding vitamin D supplementation for six- to 12-month old children. Other minerals and age groups were included in the survey to prevent bias. The response rate was 34.8%. RESULTS: Pediatricians reported higher levels (88.6%) of counseling regularly about vitamin D for infants compared to family medicine physicians (52.3%). Linear regression analysis indicated that awareness and perceived importance of vitamin D recommendations predict the frequency of vitamin D counseling regardless of specialty. CONCLUSION: Family medicine physicians in Connecticut need to increase their awareness and counseling of vitamin D supplementation for infants. PMID- 17405395 TI - Addiction and chronic pain: diagnostic and treatment dilemmas. AB - Chronic pain and addictive disorders present significant medical and public health problems; both are underdiagnosed and inadequately treated. The cooccurrence of pain and addiction adds complexity to the assessment and treatment of both disorders. Research on the link between pain and substance use disorders (SUDs) has focused on the bidirectional relationship between chronic pain and opioid dependence. While chronic opioid exposure alters physiological responsivity to painful stimuli, chronic pain also modifies the rewarding effects of opioids. Although the relationship between alcohol and pain has been less studied, alcohol use disorders are also prevalent among chronic pain patients and are of high clinical interest. In both subgroups of patients, failure to alleviate chronic pain contributes to poor clinical and functional outcomes. This review discusses common evaluation and diagnostic dilemmas in treating this challenging population. PMID- 17405396 TI - Safety of a postoperative beta-blocker infusion protocol for nonmonitored units in a community hospital. AB - OBJECTIVE: To assess the safety of a novel metoprolol infusion protocol implemented on patient care units without cardiac monitoring. DESIGN: Retrospective chart review. SETTING: 511-bed community teaching hospital. PATIENTS: 100 high cardiac-risk patients undergoing 103 noncardiac surgeries who received 354 infusions. INTERVENTION: 5, 7.5, or 10 mg of metoprolol infused over two hours. MEASUREMENTS: Infusion dosage and timing, vital signs and adverse events. RESULTS: A significant decrease in heart rate was demonstrated without any significant adverse events. CONCLUSION: Our novel metoprolol infusion protocol was safe on nonmonitored units. PMID- 17405397 TI - Neuroimaging of bilateral caudate infarction manifesting as Parkinsonian gait disorder. AB - Neurobehavioral changes characterize the clinical presentation of bilateral caudate infarction. We describe an 82-year-old man who presented with a Parkinsonian gait disorder in the absence of behavioral abnormality whose diffusion-weighted imaging demonstrated infarction of the caudate nuclei. PMID- 17405398 TI - A 58-year-old man with anti-Jo-1 syndrome and renal cell carcinoma: a case report and discussion. AB - The association of inflammatory myopathies with malignancy and interstitial lung disease has been well accepted. Polymyositis can be associated with interstitial lung disease or malignancy, although it is rare for all three entities to occur simultaneously at the initial presentation. We report a patient who was found to have concurrent polymyositis, interstitial lung disease, with anti-Jo-1 antibody, and renal cell carcinoma. PMID- 17405399 TI - Some early practitioners of Haddam. 1907. PMID- 17405400 TI - Are hospitals increasingly encroaching on the private practice of medicine. 1957. PMID- 17405401 TI - Physician-hospital relations. 1957. PMID- 17405402 TI - Doctor nurse. PMID- 17405403 TI - Connecticut's health care system. PMID- 17405404 TI - On saying goodbye to my stethoscope. PMID- 17405405 TI - Re: a "Perfect storm". PMID- 17405407 TI - Health information technology---savior or failure? PMID- 17405406 TI - Re: a "Perfect storm". PMID- 17405409 TI - Histological changes in spleens of radio-sensitive and radio-resistant mice exposed to low-dose X-ray irradiation. AB - We have previously determined by using immune-assay or bio-assay methods that low dose irradiation enhances immune and anti-oxidation functions. In this study, we examined histological changes of lymphatic follicles at 4, 24, or 48 hrs after sham, 0.25, 0.5, or 15 Gy irradiation in the spleens of BALB/c mice, which are sensitive to radiation compared with other strains, and C57BL/6J mice, which are resistant to radiation, using hematoxylin-eosin staining for lymphatic follicles or methylgreen pyronin staining for plasma cells. Results show that the lymphatic follicles in the spleens of the two mouse strains decreased at 24 or 48 hrs after 15 Gy irradiation. The number of plasma cells in the spleens of sham irradiated BALB/c mice was greater than that of sham irradiated C57BL/6J mice. At 4 hrs after 0.25 Gy irradiation, plasma cells increased in the spleens of the two mouse strains. These findings suggest, by histology, that low-dose irradiation activates the plasma cells and enhances the immune function. Although those two mouse strains have different sensitivities to radiation, the above changes were similar in both time course and degree of response. Therefore, the phenomena observed may be common in mice. PMID- 17405408 TI - Stimulation of membrane permeability transition by alpha-lipoic acid and its biochemical characteristics. AB - Mitochondria play an important role in apoptosis by generating reactive oxygen species (ROS) and inducing membrane permeability transition (MPT). Recent studies on alpha-lipoic acid (LA) and its reduced form, dihydrolipoic acid, suggest that these agents (LAs) inhibit apoptosis of cells by means of their antioxidant activity. On the other hand, LAs also stimulate Ca2+-dependent mitochondrial MPT and induce apoptosis of certain cells. Thus, the role of LAs in apoptotic cell death remains obscure. We investigated the mechanism of LA-induced MPT of mitochondria. Biochemical analysis revealed, in the presence of Ca2+, inorganic phosphate and succinate, LA induced uncoupling of oxidative phosphorylation, stimulated oxidation of pyridine nucleotides and enhanced Ca2+-induced MPT, as characterized by decrease in Ca2+ loading, ROS generation, oxidation of thiol groups of adenine nucleotide translocator, membrane depolarization, swelling, and cytochrome c release in an incubation time and concentration dependent manner. LA also stimulated hydroxyl radical-induced MPT in a alpha-tocopherol-inhibitable manner. Cyclosporine A, a potent inhibitor of mitochondrial MPT, inhibited all these events induced by LA. These results indicate that, under certain conditions, LA stimulates Ca2+-induced MPT through the decrease in loading capacity of Ca2+ and that MPT is involved in LA-induced apoptotic cell death. Since fairly high doses of LA have been used as a dietary supplement, the possible occurrence of such side effects, including mitochondrial dysfunction and induction of apoptosis in normal tissues, should be studied. PMID- 17405410 TI - Diazepam inhibits reproduction and reproductive behavior of oriental hornet. A possible role for the peripheral-type benzodiazepine receptor. AB - Feeding of diazepam to young hornets completely inhibits or delays development of their ovaries for a relatively long period. In control hornets, the ovaries usually develop within a day or two post eclosion and comb building commences on the second day of life. The hornets then oviposit into the comb cells and the deposited ova give rise to larvae. Trials were performed on parallel groups of hornets of various ages. When the sedative diazepam was administered to hornets aged 0-24 hours the ovaries of these young hornets failed to show any development, so that no oocytes ripened and consequently there was no oviposition whatsoever. Neither were any comb cells built or, at best, only a few were built. When the diazepam was administered to hornet's being the age of 48 hours, it exerted no change, that is, the eggs developed normally and comb building was the same as in the control group. Longevity of hornets was uniform in all the test groups and similar to that in the control. PMID- 17405411 TI - MRI of oriental hornet head. AB - The head of the Oriental hornet in situ, detached from a live sample was imaged using Magnetic Resonance Imaging (MRI). This non-invasive method enabled us to visualize the three-dimensional structure of the hornet's brain and intracerebral organs, as based on cubic voxels of 23 microm3. From these images, we could identify various cephalic structures in both supra-esophageal and sub-esophageal locations. In the former location, we identified and visualized the ocelli, ommatidia, mushroom body, lobula, medulla and the compound eyes in the protocerebrum, as well as the olfactory lobe and bases of the antenna in the deutocerebrum, while in the sub-esophageal region we visualized organs such as the mouthparts, the esophagus, the gnathal pouch and the salivary ducts that empty into the region. Additionally, we identified various muscles, the aorta, cuticular thickenings lending support to the interior of the head and also the cuticular skeleton providing support on the outside. All the mentioned structures and organs were visualized in their relative, normal proportions, without touching or dislocating them. PMID- 17405412 TI - In response to an open invitation for comments on AAAS project 2061's Benchmark books on science. Part 1: documentation of serious errors in cell biology. AB - Project 2061 was founded by the American Association for the Advancement of Science (AAAS) to improve secondary school science education. An in-depth study of ten 9 to 12th grade biology textbooks led to the verdict that none conveyed "Big Ideas" that would give coherence and meaning to the profusion of lavishly illustrated isolated details. However, neither the Project report itself nor the Benchmark books put out earlier by the Project carries what deserves the designation of "Big Ideas." Worse, in the two earliest-published Benchmark books, the basic unit of all life forms--the living cell--is described as a soup enclosed by a cell membrane, that determines what can enter or leave the cell. This is astonishing since extensive experimental evidence has unequivocally disproved this idea 60 years ago. A "new" version of the membrane theory brought in to replace the discredited (sieve) version is the pump model--currently taught as established truth in all high-school and college biology textbooks--was also unequivocally disproved 40 years ago. This comment is written partly in response to Bechmark's gracious open invitation for ideas to improve the books and through them, to improve US secondary school science education. PMID- 17405413 TI - Five successful methods to control spending on physician preference items. PMID- 17405414 TI - Supply execs expect price hikes to continue in 2007. PMID- 17405416 TI - Develop vendor management policy to control new products. PMID- 17405415 TI - Collaboration is the key to effective expense management. PMID- 17405417 TI - Radiation dose in computed tomography: why it's a concern and what you can do about it. AB - Computed tomography (CT) has become an invaluable and even life-saving diagnostic tool. But its great benefits come at a price: Because it uses ionizing radiation, CT can cause cancer. In fact, in the United States alone, it is estimated that CT accounts for around 6,000 additional cancers per year, with about half of those proving fatal. Unfortunately, this risk is not as widely recognized as it should be. In particular, both clinicians and patients tend to lack a clear understanding of the radiation doses involved in CT studies, which are higher than those used in other common x-ray studies. Given the swiftly growing use of CT, there's the real possibility that patients' exposure to radiation will grow as well. Thus, it's vital that clinicians understand the doses involved in CT, that those doses be kept as low as possible, and that CT be used only when the clinical benefits justify it. In this article, we explain the radiation risks of CT and discuss how they compare with other risks in and out of healthcare. We also discuss ways in which the doses delivered to patients can be kept to the necessary minimum. The approaches we cover include eliminating unnecessary CT use, raising referring physicians' awareness of the problem, considering alternative technologies, using good quality assurance, and optimizing imaging protocols. PMID- 17405418 TI - Problem reports. Manufacturer policy changes push Mallinckrodt Liebel-Flarsheim CT9000 ADV customers toward employing used-syringe safety feature. PMID- 17405419 TI - Cost differences between academic and nonacademic hospitals: a case study of surgical procedures. AB - Decreased public funding, a competitive healthcare market, and higher patient care costs have been blamed for the present financial challenges that confront academic health centers. The authors examined the costs associated with graduate medical education, particularly, indirect medical education expenses in the operating room. The results indicate that it is more costly for teaching hospitals to provide surgical care to patients in the operating room. The academic health center's indirect graduate medical expenses only covered a portion of the increased costs. If the missions of academic health centers are perceived as a public good, policy makers must design a system that more appropriately compensates academic health centers for the additional costs associated with surgical procedures in graduate medical education. PMID- 17405420 TI - Hospital strategic planning diversity integration based organizational type and CEO tenure. AB - The author investigated the strategic planning processes used by five different hospital types to integrate diversity practices into their operations. Chief executive officers from Delaware, New Jersey, New York, and Pennsylvania were surveyed to examine how their hospitals used strategic planning in this process. The central research question was: Does this process vary on the basis of hospital type? The findings indicated that some differences do exist by hospital type, as well as the length of tenure for CEOs in their positions. PMID- 17405421 TI - Five constants of information technology adoption in healthcare. AB - The healthcare industry has developed a dependence on information technology (IT) for maintaining and improving both clinical and business operations. Whether IT is used for office automation or for reducing medical errors, there are five constants that routinely influence the successful integration of IT in healthcare. These constants are the proper use and maintenance of the IT budget, the role of supportive leadership, the use of project management, the process of implementation, and the significance of end user involvement. These constants challenge healthcare organizations to efficiently and effectively use their financial and human resources when adopting new IT. These constants also shape how the healthcare industry approaches the adoption and utilization of new IT. A collective understanding of these constants and their interrelationships will enable healthcare organizations to better integrate new IT and achieve organizational goals of developing a solid technological infrastructure to truly enhance the delivery of quality healthcare. PMID- 17405422 TI - Disparities in access to healthcare: the case of a drug and alcohol abuse detoxification treatment program among minority groups in a Texas hospital. AB - The authors analyzed ethnic/racial disparities in healthcare access and length of stay from a defined population of individuals seeking medical detoxification services at a hospital in Texas. Results indicated Blacks were more likely to be insured compared with Whites, mostly by public insurance, but this did not hold for Hispanics, who were about three times more likely to be uninsured compared with Blacks. In addition, the authors observed lower median of length of stay in the Medicaid category among Hispanics. These results can be explained by aggressive case management, sociocultural barriers, or discriminatory practices, both intentional and unintentional. PMID- 17405423 TI - Assistance in dying: part I. Europe--the vanguard. AB - This Nexus provides background information about assistance in dying. The international perspective based on developments in Western Europe, especially the Netherlands, provides a useful context in which to place events in the United States. Part II considers the history and legal context of assistance in dying in the United States. Oregon is given special attention because it remains the only state with legally sanctioned assistance in dying. Readers are urged to bear in mind the definitions and distinctions among assisted dying, the various forms of euthanasia, and physician-assisted suicide. Informed observers of the discussion will readily see that the terms and concepts are often used imprecisely and as though they are synonymous. Productive debate of assistance in dying will occur only if all those involved agree on definitions and the meaning of the concepts. PMID- 17405424 TI - A new orientation-adaptive interpolation method. AB - We propose an isophote-oriented, orientation-adaptive interpolation method. The proposed method employs an interpolation kernel that adapts to the local orientation of isophotes, and the pixel values are obtained through an oriented, bilinear interpolation. We show that, by doing so, the curvature of the interpolated isophotes is reduced, and, thus, zigzagging artifacts are largely suppressed. Analysis and experiments show that images interpolated using the proposed method are visually pleasing and almost artifact free. PMID- 17405425 TI - State-space analysis of cardiac motion with biomechanical constraints. AB - Quantitative estimation of nonrigid motion from image sequences has important technical and practical significance. State-space analysis provides powerful and convenient ways to construct and incorporate the physically meaningful system dynamics of an object, the image-derived observations, and the process and measurement noise disturbances. In this paper, we present a biomechanical-model constrained state-space analysis framework for the multiframe estimation of the periodic cardiac motion and deformation. The physical constraints take the roles as spatial regulator of the myocardial behavior and spatial filter/interpolator of the data measurements, while techniques from statistical filtering theory impose spatiotemporal constraints to facilitate the incorporation of multiframe information to generate optimal estimates of the heart kinematics. Physiologically meaningful results have been achieved from estimated displacement fields and strain maps using in vivo left ventricular magnetic resonance tagging and phase contrast image sequences, which provide the tag-tag and tag-boundary displacement inputs, and the mid-wall instantaneous velocity information and boundary displacement measures, respectively. PMID- 17405426 TI - Multidimensional directional filter banks and surfacelets. AB - In 1992, Bamberger and Smith proposed the directional filter bank (DFB) for an efficient directional decomposition of 2-D signals. Due to the nonseparable nature of the system, extending the DFB to higher dimensions while still retaining its attractive features is a challenging and previously unsolved problem. We propose a new family of filter banks, named NDFB, that can achieve the directional decomposition of arbitrary N-dimensional (N > or =2) signals with a simple and efficient tree-structured construction. In 3-D, the ideal passbands of the proposed NDFB are rectangular-based pyramids radiating out from the origin at different orientations and tiling the entire frequency space. The proposed NDFB achieves perfect reconstruction via an iterated filter bank with a redundancy factor of N in N-D. The angular resolution of the proposed NDFB can be iteratively refined by invoking more levels of decomposition through a simple expansion rule. By combining the NDFB with a new multiscale pyramid, we propose the surfacelet transform, which can be used to efficiently capture and represent surface-like singularities in multidimensional data. PMID- 17405427 TI - Tomographic reconstruction of dynamic cardiac image sequences. AB - In this paper, we propose an approach for the reconstruction of dynamic images from a gated cardiac data acquisition. The goal is to obtain an image sequence that can show simultaneously both cardiac motion and time-varying image activities. To account for the cardiac motion, the cardiac cycle is divided into a number of gate intervals, and a time-varying image function is reconstructed for each gate. In addition, to cope with the under-determined nature of the problem, the time evolution at each pixel is modeled by a B-spline function. The dynamic images for the different gates are then jointly determined using maximum a posteriori estimation, in which a motion-compensated smoothing prior is introduced to exploit the similarity among the different gates. The proposed algorithm is evaluated using a dynamic version of the 4-D gated mathematical cardiac torso phantom simulating a gated single photon emission computed tomography perfusion acquisition with Technitium-99m labeled Teboroxime. We thoroughly evaluated the performance of the proposed algorithm using several quantitative measures, including signal-to-noise ratio analysis, bias-variance plot, and time activity curves. Our results demonstrate that the proposed joint reconstruction approach can improve significantly the accuracy of the reconstruction. PMID- 17405428 TI - Quality evaluation of motion-compensated edge artifacts in compressed video. AB - Little attention has been paid to an impairment common in motion-compensated video compression: the addition of high-frequency (HF) energy as motion compensation displaces blocking artifacts off block boundaries. In this paper, we employ an energy-based approach to measure this motion-compensated edge artifact, using both compressed bitstream information and decoded pixels. We evaluate the performance of our proposed metric, along with several blocking and blurring metrics, on compressed video in two ways. First, ordinal scales are evaluated through a series of expectations that a good quality metric should satisfy: the objective evaluation. Then, the best performing metrics are subjectively evaluated. The same subjective data set is finally used to obtain interval scales to gain more insight. Experimental results show that we accurately estimate the percentage of the added HF energy in compressed video. PMID- 17405430 TI - Efficient entropy estimation based on doubly stochastic models for quantized wavelet image data. AB - Under a rate constraint, wavelet-based image coding involves strategic discarding of information such that the remaining data can be described with a given amount of rate. In a practical coding system, this task requires knowledge of the relationship between quantization step size and compressed rate for each group of wavelet coefficients, the R-Q curve. A common approach to this problem is to fit each subband with a scalar probability distribution and compute entropy estimates based on the model. This approach is not effective at rates below 1.0 bits-per pixel because the distributions of quantized data do not reflect the dependencies in coefficient magnitudes. These dependencies can be addressed with doubly stochastic models, which have been previously proposed to characterize more localized behavior, though there are tradeoffs between storage, computation time, and accuracy. Using a doubly stochastic generalized Gaussian model, it is demonstrated that the relationship between step size and rate is accurately described by a low degree polynomial in the logarithm of the step size. Based on this observation, an entropy estimation scheme is presented which offers an excellent tradeoff between speed and accuracy; after a simple data-gathering step, estimates are computed instantaneously by evaluating a single polynomial for each group of wavelet coefficients quantized with the same step size. These estimates are on average within 3% of a desired target rate for several of state of-the-art coders. PMID- 17405429 TI - A new divergence measure for medical image registration. AB - A new type of divergence measure for the registration of medical images is introduced that exploits the properties of the modified Bessel functions of the second kind. The properties of the proposed divergence coefficient are analysed and compared with those of the classic measures, including Kullback-Leibler, Renyi, and Iinfinity, divergences. To ensure its effectiveness and widespread applicability to any arbitrary set of data types, the performance of the new measure is analysed for Gaussian, exponential, and other advanced probability density functions. The results verify its robustness. Finally, the new divergence measure is used in the registration of CT to MR medical images to validate the improvement in registration accuracy. PMID- 17405431 TI - Ordering for embedded coding of wavelet image data based on arbitrary scalar quantization schemes. AB - Many modern wavelet quantization schemes specify wavelet coefficient step sizes as continuous functions of an input step-size selection criterion; rate control is achieved by selecting an appropriate set of step sizes. In embedded wavelet coders, however, rate control is achieved simply by truncating the coded bit stream at the desired rate. The order in which wavelet data are coded implicitly controls quantization step sizes applied to create the reconstructed image. Since these step sizes are effectively discontinuous, piecewise-constant functions of rate, this paper examines the problem of designing a coding order for such a coder, guided by a quantization scheme where step sizes evolve continuously with rate. In particular, it formulates an optimization problem that minimizes the average relative difference between the piecewise-constant implicit step sizes associated with a layered coding strategy and the smooth step sizes given by a quantization scheme. The solution to this problem implies a coding order. Elegant, near-optimal solutions are presented to optimize step sizes over a variety of regions of rates, either continuous or discrete. This method can be used to create layers of coded data using any scalar quantization scheme combined with any wavelet bit-plane coder. It is illustrated using a variety of state-of the-art coders and quantization schemes. In addition, the proposed method is verified with objective and subjective testing. PMID- 17405432 TI - Lossless video sequence compression using adaptive prediction. AB - We present an adaptive lossless video compression algorithm based on predictive coding. The proposed algorithm exploits temporal, spatial, and spectral redundancies in a backward adaptive fashion with extremely low side information. The computational complexity is further reduced by using a caching strategy. We also study the relationship between the operational domain for the coder (wavelet or spatial) and the amount of temporal and spatial redundancy in the sequence being encoded. Experimental results show that the proposed scheme provides significant improvements in compression efficiencies. PMID- 17405433 TI - A maximum entropy approach to unsupervised mixed-pixel decomposition. AB - Due to the wide existence of mixed pixels, the derivation of constituent components (endmembers) and their fractional proportions (abundances) at the subpixel scale has been given a lot of attention. The entire process is often referred to as mixed-pixel decomposition or spectral unmixing. Although various algorithms have been proposed to solve this problem, two potential issues still need to be further investigated. First, assuming the endmembers are known, the abundance estimation is commonly performed by employing a least-squares error criterion, which, however, makes the estimation sensitive to noise and outliers. Second, the mathematical intractability of the abundance non-negative constraint results in computationally expensive numerical approaches. In this paper, we propose an unsupervised decomposition method based on the classic maximum entropy principle, termed the gradient descent maximum entropy (GDME), aiming at robust and effective estimates. We address the importance of the maximum entropy principle for mixed-pixel decomposition from a geometric point of view and demonstrate that when the given data present strong noise or when the endmember signatures are close to each other, the proposed method has the potential of providing more accurate estimates than the popular least-squares methods (e.g., fully constrained least squares). We apply the proposed GDME to the subject of unmixing multispectral and hyperspectral data. The experimental results obtained from both simulated and real images show the effectiveness of the proposed method. PMID- 17405434 TI - Combined error concealment and error correction in rate-distortion analysis for multiple substream transmissions. AB - We propose a new framework for multiple scalable bitstream video communications over lossy channels. The major feature of the framework is that the encoder estimates the effects of postprocessing concealment and includes those effects in the rate-distortion analysis. Based on the framework, we develop a rate distortion optimization algorithm to generate multiple scalable bitstreams. The algorithm maximizes the expected peak signal-to-noise ratio by optimally assigning forward error control codes and transmission schemes in a constrained bandwidth. The framework is a general approach motivated by previous methods that perform concealment in the decoder, as in our special case. Simulations show that the proposed approach can be implemented efficiently and that it outperforms previous methods by more than 2 dB. PMID- 17405435 TI - Vertex-based diffusion for 3-D mesh denoising. AB - We present a vertex-based diffusion for 3-D mesh denoising by solving a nonlinear discrete partial differential equation. The core idea behind our proposed technique is to use geometric insight in helping construct an efficient and fast 3-D mesh smoothing strategy to fully preserve the geometric structure of the data. Illustrating experimental results demonstrate a much improved performance of the proposed approach in comparison with existing methods currently used in 3 D mesh smoothing. PMID- 17405436 TI - Interactive image segmentation via adaptive weighted distances. AB - An interactive algorithm for soft segmentation of natural images is presented in this paper. The user first roughly scribbles different regions of interest, and from them, the whole image is automatically segmented. This soft segmentation is obtained via fast, linear complexity computation of weighted distances to the user-provided scribbles. The adaptive weights are obtained from a series of Gabor filters, and are automatically computed according to the ability of each single filter to discriminate between the selected regions of interest. We present the underlying framework and examples showing the capability of the algorithm to segment diverse images. PMID- 17405437 TI - Perceptual color correction through variational techniques. AB - In this paper, we present a discussion about perceptual-based color correction of digital images in the framework of variational techniques. We propose a novel image functional whose minimization produces a perceptually inspired color enhanced version of the original. The variational formulation permits a more flexible local control of contrast adjustment and attachment to data. We show that a numerical implementation of the gradient descent technique applied to this energy functional coincides with the equation of automatic color enhancement (ACE), a particular perceptual-based model of color enhancement. Moreover, we prove that a numerical approximation of the Euler-Lagrange equation reduces the computational complexity of ACE from theta(N2) to theta(N log N), where N is the total number of pixels in the image. PMID- 17405438 TI - Fast and stable bayesian image expansion using sparse edge priors. AB - Smoothness assumptions in traditional image expansion cause blurring of edges and other high-frequency content that can be perceptually disturbing. Previous edge preserving approaches are either ad hoc, statistically untenable, or computationally unattractive. We propose a new edge-driven stochastic prior image model and obtain the maximum a posteriori (MAP) estimate under this model. The MAP estimate is computationally challenging since it involves the inversion of very large matrices. An efficient algorithm is presented for expansion by dyadic factors. The technique exploits diagonalization of convolutional operators under the Fourier transform, and the sparsity of our edge prior, to speed up processing. Visual and quantitative comparison of our technique with other popular methods demonstrates its potential and promise. PMID- 17405439 TI - Binary weighted averaging of an ensemble of coherently collected image frames. AB - Recent interest in the collection of remote laser radar imagery has motivated novel systems that process temporally contiguous frames of collected imagery to produce an average image that reduces laser speckle, increases image SNR, decreases the deleterious effects of atmospheric distortion, and enhances image detail. This research seeks an algorithm based on Bayesian estimation theory to select those frames from an ensemble that increases spatial resolution compared to simple unweighted averaging of all frames. The resulting binary weighted motion-compensated frame average is compared to the unweighted average using simulated and experimental data collected from a fielded laser vision system. Image resolution is significantly enhanced as quantified by the estimation of the atmospheric seeing parameter through which the average image was formed. PMID- 17405440 TI - Deblurring of color images corrupted by impulsive noise. AB - We consider the problem of restoring a multichannel image corrupted by blur and impulsive noise (e.g., salt-and-pepper noise). Using the variational framework, we consider the L1 fidelity term and several possible regularizers. In particular, we use generalizations of the Mumford-Shah (MS) functional to color images and gamma-convergence approximations to unify deblurring and denoising. Experimental comparisons show that the MS stabilizer yields better results with respect to Beltrami and total variation regularizers. Color edge detection is a beneficial by-product of our methods. PMID- 17405441 TI - A detection statistic for random-valued impulse noise. AB - This paper proposes an image statistic for detecting random-valued impulse noise. By this statistic, we can identify most of the noisy pixels in the corrupted images. Combining it with an edge-preserving regularization, we obtain a powerful two-stage method for denoising random-valued impulse noise, even for noise levels as high as 60%. Simulation results show that our method is significantly better than a number of existing techniques in terms of image restoration and noise detection. PMID- 17405442 TI - A class-adaptive spatially variant mixture model for image segmentation. AB - We propose a new approach for image segmentation based on a hierarchical and spatially variant mixture model. According to this model, the pixel labels are random variables and a smoothness prior is imposed on them. The main novelty of this work is a new family of smoothness priors for the label probabilities in spatially variant mixture models. These Gauss-Markov random field-based priors allow all their parameters to be estimated in closed form via the maximum a posteriori (MAP) estimation using the expectation-maximization methodology. Thus, it is possible to introduce priors with multiple parameters that adapt to different aspects of the data. Numerical experiments are presented where the proposed MAP algorithms were tested in various image segmentation scenarios. These experiments demonstrate that the proposed segmentation scheme compares favorably to both standard and previous spatially constrained mixture model-based segmentation. PMID- 17405443 TI - Binary partition tree analysis based on region evolution and its application to tree simplification. AB - Pyramid image representations via tree structures are recognized methods for region-based image analysis. Binary partition trees can be applied which document the merging process with small details found at the bottom levels and larger ones close to the root. Hindsight of the merging process is stored within the tree structure and provides the change histories of an image property from the leaf to the root node. In this work, the change histories are modelled by evolvement functions and their second order statistics are analyzed by using a knee function. Knee values show the reluctancy of each merge. We have systematically formulated these findings to provide a novel framework for binary partition tree analysis, where tree simplification is demonstrated. Based on an evolvement function, for each upward path in a tree, the tree node associated with the first reluctant merge is considered as a pruning candidate. The result is a simplified version providing a reduced solution space and still complying with the definition of a binary tree. The experiments show that image details are preserved whilst the number of nodes is dramatically reduced. An image filtering tool also results which preserves object boundaries and has applications for segmentation. PMID- 17405444 TI - A coupled statistical model for face shape recovery from brightness images. AB - We focus on the problem of developing a coupled statistical model that can be used to recover facial shape from brightness images of faces. We study three alternative representations for facial shape. These are the surface height function, the surface gradient, and a Fourier basis representation. We jointly capture variations in intensity and the surface shape representations using a coupled statistical model. The model is constructed by performing principal components analysis on sets of parameters describing the contents of the intensity images and the facial shape representations. By fitting the coupled model to intensity data, facial shape is implicitly recovered from the shape parameters. Experiments show that the coupled model is able to generate accurate shape from out-of-training-sample intensity images. PMID- 17405445 TI - A new family of nonredundant transforms using hybrid wavelets and directional filter banks. AB - We propose a new family of nonredundant geometrical image transforms that are based on wavelets and directional filter banks. We convert the wavelet basis functions in the finest scales to a flexible and rich set of directional basis elements by employing directional filter banks, where we form a nonredundant transform family, which exhibits both directional and nondirectional basis functions. We demonstrate the potential of the proposed transforms using nonlinear approximation. In addition, we employ the proposed family in two key image processing applications, image coding and denoising, and show its efficiency for these applications. PMID- 17405446 TI - Semantic-based surveillance video retrieval. AB - Visual surveillance produces large amounts of video data. Effective indexing and retrieval from surveillance video databases are very important. Although there are many ways to represent the content of video clips in current video retrieval algorithms, there still exists a semantic gap between users and retrieval systems. Visual surveillance systems supply a platform for investigating semantic based video retrieval. In this paper, a semantic-based video retrieval framework for visual surveillance is proposed. A cluster-based tracking algorithm is developed to acquire motion trajectories. The trajectories are then clustered hierarchically using the spatial and temporal information, to learn activity models. A hierarchical structure of semantic indexing and retrieval of object activities, where each individual activity automatically inherits all the semantic descriptions of the activity model to which it belongs, is proposed for accessing video clips and individual objects at the semantic level. The proposed retrieval framework supports various queries including queries by keywords, multiple object queries, and queries by sketch. For multiple object queries, succession and simultaneity restrictions, together with depth and breadth first orders, are considered. For sketch-based queries, a method for matching trajectories drawn by users to spatial trajectories is proposed. The effectiveness and efficiency of our framework are tested in a crowded traffic scene. PMID- 17405447 TI - A comment on "Asymptotically optimal detection for additive watermarking in the DCT AND DWT domains". AB - Explicit expressions are derived for the Fisher information for the generalized Gaussian noise model considered by Nikolaidis and Pitas. PMID- 17405448 TI - Thinking outside the pyramidal cell: unexplored contributions of interneurons and neuropeptide Y to estrogen-induced synapse formation in the hippocampus. AB - Since the first finding that 17beta-estradiol (E) can regulate CA1 pyramidal cell synapse formation, subsequent studies have explored many potential E-dependent mechanisms occurring within CA1 pyramidal cells. Fewer studies have focused on E dependent processes outside of the pyramidal cell that may influence events activity of the pyramidal cells. This review considers hippocampal interneurons, which can potently regulate the excitability of simultaneously firing pyramidal cells. In particular, we discuss neuropeptide Y (NPY) expression by these interneurons because our published findings show that NPY expression is increased by E in a subset of interneurons which coincidentally exhibit E-regulated increase in GABA synthesis and are uniquely situated anatomically such that they may regulate synaptic activity. Here we review the role of different phenotypes of CA1 interneurons, and we propose a model in which E-stimulated NPY gene expression and the release of NPY by interneurons inhibits glutamate release presynaptically and alters glutamate-dependent synaptic events in the rat hippocampus during adulthood. PMID- 17405449 TI - Electrical synapses in basal ganglia. AB - The basal ganglia (BG) provide a major integrative system of the forebrain involved in the organization of goal-directed behaviour. Pathological alteration of BG function leads to major motor and cognitive impairments such as observed in Parkinson's disease. Recent advances in BG research stress the role of neural oscillations and synchronization in the normal and pathological function of BG. As demonstrated in several brain structures, these patterns of neural activity can emerge from electrically coupled neuronal networks. This review aims at addressing the presence, functionality and putative role of electrical synapses in BG, with a particular emphasis on the striatum and the substantia nigra pars compacta (SNc), two main BG nuclei in which the existence and functional properties of neuronal coupling are best documented. PMID- 17405450 TI - Insights into the role of dopamine receptor systems in learning and memory. AB - It is well established that learning and memory are complex processes involving and recruiting different brain modulatory neurotransmitter systems. Considerable evidence points to the involvement of dopamine in various aspects of cognition, and interest has been focused on investigating the clinical relevance of dopamine systems to age-related cognitive decline and manifestations of cognitive impairment in schizophrenia, Alzheimer's disease, Parkinson's disease and other neurodegenerative diseases. In the past decade or so, in spite of the molecular cloning of the five dopamine receptor subtypes, their specific roles in brain function remained inconclusive due to the lack of completely selective ligands that could distinguish between the members of the D1-like and D2-like dopamine receptor families. One of the most important advances in the field of dopamine research has been the generation of mutant mouse models permitting evaluation of the dopaminergic system using gene targeting technologies. These mouse models represent an important approach to explore the functional roles of closely related receptor subtypes. In this review, we present and discuss evidence on the role of dopamine receptors in different aspects of learning and memory at the cellular, molecular and behavioral levels. We compare evidence using conventional pharmacological, lesion or electrophysiological studies with results from mice with targeted deletions of different subtypes of dopamine receptor genes. We particularly focus on dopamine D1 and D2 receptors in an effort to delineate their specific roles in various aspects of cognitive function. We provide strong evidence, from our own recent work as well as others, that dopamine is part of the network that plays a very important role in cognitive function, and that although multiple dopamine receptor subtypes contribute to different aspects of learning and memory, the D1 receptor seems to play a more prominent role in mediating plasticity and specific aspects of cognitive function, including spatial learning and memory processes, reversal learning, extinction learning, and incentive learning. PMID- 17405451 TI - Adult hippocampal neurogenesis in rodents and primates: endogenous, enhanced, and engrafted. AB - Since the studies of Ramon y Cajal, a central postulate in neuroscience has been the view that the adult brain lacks the ability to regenerate its neurons. This dogma has been challenged in the last few decades, and mounting evidence has accumulated showing the existence of a phenomenon designated 'adult neurogenesis'. De novo generation of neurons by neural progenitor cells in the adult brain is thought to be preserved only in restricted brain areas, such as the hippocampal dentate gyrus (DG) and the subventricular zone (SVZ) of the lateral ventricle. Data in the last decade coming mostly from rodent models have clearly documented that precursor cells residing in the anterior portion of SVZ and the subgranular zone of DG are responsible for adding new neurons in the olfactory bulb and DG, respectively. This raised significant interest in the clinical potential of neural progenitor cells, and recent studies have documented that brain injury is capable of activating an endogenous program of neurogenesis resulting in neuronal replacement in various cerebral regions of rodents and primates. If the newly generated neurons in the adult brain prove to be functional, it could have a tremendous impact for cell replacement therapies. Here, we summarize current knowledge of the mechanisms affecting adult hippocampal neurogenesis in both rodents and primates, and discuss its implications in developing novel strategies for the treatment of human neurological diseases. PMID- 17405452 TI - Genetic susceptibility and development of chronic non-malignant back pain. AB - Previous data indicate that persistent pain states often involve sensitization within the central nervous system (CNS). Many recently described human genetic variants may affect these central processes. Genetic variability influences both synthesis and function of proteins affecting the plasticity of the CNS. Hence, individual genetic variability may be important to understand the development of many persistent pain conditions including chronic nonmalignant back pain. In this review we argue that genotyping of each patient may be a valuable complement to diagnosis of back disorders. This may be important for future prescription of medicine to individuals predisposed for persistent pain. Increased understanding of genetic variability may also improve multidisciplinary and cognitive behavioral approaches to management of persistent pain. Translation of this information from the laboratory into clinical application will be important for future prevention as well as treatment of long-lasting non-malignant pain conditions. PMID- 17405453 TI - [Sudden cardiorespiratory arrest in adults with acute epiglottitis. Report of 2 cases]. AB - We report two adult cases of sudden cardiorespiratory arrest because an acute epiglottitis that ocurred in our hospital. Although it is an uncommon pathology nowdays with good prognosis generally, we try to call the attention about this complication which can happen of unforseeable way even in previous absence of respiratory obstruction. One of the patients required coniotomy in ICU due to impossibility of orotracheal intubation (OI) and the other died in spite of an urgent performance. PMID- 17405454 TI - [Spontaneous cervical and pharyngolaryngeal hematoma after coughing attacks]. AB - We report the case of a 86 years old male who, in the course of a coughing attack, refers spontaneous cervical and pharyngeal pain in addition to hoarness, dysphagia and some dyspnea. His exploration revealed a small right neck hematoma but with infiltration of the orohypopharynx and right hemilarynx for spreading later to posterior pharyngeal wall and left aryepiglottic fold. The patient was sent to ICU although he did not required a control of airway by intubation or tracheotomy finally. PMID- 17405455 TI - [Laryngeal leishmaniasis]. AB - Leishmaniasis consists in a zoonotic infection, that means affect animals. Due to the puncture of the female sand fly type Phlebotomus (Ancient World) or Lutzomya (New World) the human being can be also affected. The clinical manifestations are very varied, depending to factors related with host, vector and the proper parasit. In this present work, we expose the three unic cases of Leishmaniasis affecting the larynx, diagnosticated in the sanitary area of Toledo. Due to the specific characteristics that present each case different treatments for each one have been realised, and its results and treatment exposed. PMID- 17405456 TI - [Cervical myositis ossificans. A case report]. AB - This is the case of a middle-aged male with a slow growing lateral cervcal painful tumour, and without previous history of local trauma. Radiological testing reveals an ossifying soft tissue lesion without any osseous compromise. The lesion is completely resected, and the histological result is of ossifying myositis. It is an osseous tumour non neoplastic that originates within muscle and in particular the flexors of the inferior extremities and thigs or in the soft tissues. The incidence within the head and neck region is low (20%). There is an hereditary progressive form, Munchmeyer's disease, and another circumscribed, which can be subdivided in traumatic or atraumatic. PMID- 17405457 TI - Cranial base fracture and rhino cerebrospinmal fluid leakage. A case report. AB - Oto cerebrospinal fluid leakage occurs frequently in skull base fractures but it is not always recognized which may produce potentially serious consequences on the prognosis. The aim of this study is to present a case of an extended skull base fracture with bad defined symptoms. A male in coma was admitted to our hospital following a road accident. Imaging revealed a fracture that transversally crossed the squamous occipital bone and petrous portions of temporal bone on the right, the sphenoid bone, and the left zygomatic bone. Ten days later the patient regained consciousness presenting symptoms of right complete hearing loss, cephalalgia and fever. Lumbar puncture showed a Gram negative germ growth. After specific antibiotic treatment he underwent surgery with exclusion of the middle ear and the mastoid from outside by obliterating the Eustachian tube, sealing the surgical cavity (subtotal petrosectomy) with abdominal fat and closing the external auditory canal as a blind sac. At the same time, nasal fibroendoscopy was also performed to close the fistulas in the sphenoid region. Since the cephalalgia persisted further CT examination was performed and revealed another fracture rima in the ethmoid bone. Nasal fibroendoscopy was performed again to close this fistula. The symptoms thus disappeared and the patient has continued to be symptom-free during the two years follow-up. Skull base fractures may involve various bone structure (petrous portion of temporal bone, ethmoid, sphenoid, parietal bone). As a result of the complex anatomy of the skull base, the fracture may damage numerous vital structures (cranial nerves, internal carotid artery, cavernous sinus, jugular vein etc) and the dura mater, causing cerebrospinal fluid leak. When the fracture in the petrous bone is transversal, it is highly important not to delay surgery. In fact the otic capsule does not repair but the bone step is covered by a thin layer of fibrous tissue. For this reason patients, with clear damage to the otic capsule, risk meningitis. Fistulas in the ethmoid are the most difficult to diagnose and the easiest to underestimate. It is fundamental to follow the appropriate diagnostic procedure. PMID- 17405458 TI - [Pseudomonas and acute external otitis. Results of a microbiological study in patients without previous antibiotic treatment]. AB - We report the results of a microbiological clinic study that was performed by our ENT Department between years 2000 and 2001 whose main objective was to determine, in Badajoz Area of Health, which bacteria were involved in the acute diffuse external otitis of patients without previous antibiotic treatment (two weeks before obtaining the samples). Of 79 isolated microorganisms in 62 patients that fulfilled the requirements established Pseudomonas, mainly P. Aeruginosa, represented a 46.83% altogether followed by Staphylococcus (18.98%). In almost one fourth part of the cases strains of associated fungi were identified. PMID- 17405459 TI - [Supracricoid partial laryngectomy: quality of life assessment]. AB - OBJECTIVES: To assess quality of life in patients diagnosed with epidermoid carcinoma of the larynx, after supracricoid partial laryngectomy (SCPL). To examine how quality-of-life may be associated to the treatment received. PATIENTS AND METHOD: A descriptive, observational, cross-sectional study was conducted involving 32 patients out of a group of 57. They were all males with an average age of 63.3 years at the time of the interview. We used the disease-specific quality of life questionnaire devised by the University of Michigan: the Head and Neck Cancer-specific Quality of life instrument (HNQOL). RESULTS: During an average follow-up period of 86 months, 3 patients (5.26%) suffered a recurrence. Estimated survival after 3, 5 and 10 years was 94.73%. Communication and overall bother caused by treatment were found to be the two quality-of-life domains mainly affected. Patients who had received CHP, radiotherapy, those who had not had their tracheal cannula removed or had undergone neck dissection were the most affected. CONCLUSIONS: SCPL as a surgical technique allows good control of the condition and has a low impact on the patient's quality of life. PMID- 17405460 TI - [Treatment of the benign paroxysmal positional vertigo (BPPV) by Epley maneuver]. AB - Benign paroxysmal positional vertigo (BPPV) is the most common of the peripheral vertigo. We report the case of a 45 years old female with that pathology suspected by the symptoms which the patient related to head changes of positions and movements. Her diagnosis was confirmed by the maneuver of Dix-Hallpike, that was positive, and the treatment consisted in a maneuver of canalicular reposition according to Epley's technique. Such maneuver was successful and actually the patient is without symptoms after more than 2 years since the first episode. Finally we have performed a bibliographic review to verify the effectiveness of that treatment. PMID- 17405461 TI - [Otorhinolaringologyc approach of the chronic cough. Clinical case]. AB - Chronic cough is defined as that which is persisting at least for trhee weeks without an evident cause. It is very common on the otorhinological practice to receive patients with chronic cough in order to rule out that their chronic cough is not produced because of an alteration in the high respiratory system. We show a clinical case with chronic cough and we make reference to the physical exploration, diagnostical method, and possibilities of medical and surgical treatment. PMID- 17405462 TI - [Recurrent intrathyroid abscess in children. Clinical case and review of the literature]. AB - Recurrent thyroid abscesses in children are an uncommon clinical disorder that use to be due to 3rd or 4th branchial anomalies and pyriform sinus fistula. We are reporting the clinical case ofa 6 year-girl who was diagnosed as left thyroid lobe and istmic abscess of 3 cm, drained by general anesthesia with positive culture for St. Viridans. Four months later she was re-admitted by the same cause and we performed a new drainage, by punction, and endovenous antimicrobial therapy for aerobic and anaerobic pathogens with good evolution. The imaging tests showed no anatomical alterations. PMID- 17405463 TI - Expanding dental undergraduate clinical outreach programmes into general dental practice. PMID- 17405464 TI - Community effectiveness of fissure sealants and the effect of fluoridated water consumption. AB - OBJECTIVES: This study investigated the effectiveness of fissure sealants in a community-based programme and the relationship between fissure sealants effectiveness and exposure to fluoridated water. DESIGN: Prospective cohort study of children attending the School Dental Service (SDS) of two Australian states, Queensland and South Australia, across a period of between 6 months and 3.5 years (mean = 2 years). METHODS: Oral health data on 4-15-year-olds were obtained as part of regular examinations by the SDS while questionnaire data on residential and water consumption history were provided by children's parents or guardians. PARTICIPANTS: A sub-group of 789 children (mean age = 10.5 years) was selected with one contralateral pair of permanent first molars at baseline where the occlusal surface of one molar had been fissure sealed while the paired surface was diagnosed as sound. RESULTS: The caries incidence of the fissure sealed occlusal surfaces was 5.6% compared to 11.1% for sound surfaces (p < 0.001), demonstrating a 50% reduction in caries incidence for sealed vs non-sealed surfaces. The reduction in caries increment attributable to fissure sealing increased across fluoridated water exposure categories--a 36.4% reduction was found for children with 0% exposure (p > 0.05), a 55.0% reduction for children with intermediate exposure (p < 0.01), and an 82.4% reduction for children with 100% lifetime exposure to fluoridated water (p < 0.001). Differences between odds ratios for fissure sealants across exposure categories were not statistically significant. CONCLUSIONS: The effectiveness of fissure sealants in community based programmes may be further improved when coupled with increased lifetime exposure to optimally fluoridated water. PMID- 17405465 TI - Financial systems' impact on dental care; a review of fee-for-service and capitation systems. AB - OBJECTIVE: This review covers the impact of financial systems on dental care. BACKGROUND: Remuneration in fee-for-service (FFS) is done per service provided and in capitation (CAP) per patient enrolled. It may be expected that dentists' incentive in CAP is to keep the number of services provided at a minimum, while in FFS it is to keep the number of services at a maximum. This should lead to a different impact on care, with the dentists in CAP focusing more on prevention and dentists in FFS more on restorative treatment. Six questions were put: Does CAP increase or decrease caries incidence? Does CAP increase or decrease restorative treatments? Does CAP increase preventive care? Does CAP increase or decrease productivity? Does CAP increase or decrease the dentist's satisfaction with his/her work? Does CAP increase or decrease the patients' satisfaction with the oral care provided? METHODS: Literature was obtained through searches in databases. A format was developed to define the literature of interest. RESULTS: CAP decreases restorative treatment and there is a tendency of decreased caries incidence. "Supervised neglect" cannot be established. CAP increases preventive care. A conclusion regarding productivity was not possible. The results on dentist's satisfaction with work were inconclusive, as were the results regarding patient satisfaction. CONCLUSIONS: CAP has a different impact on provided care than FFS. More research is needed in this area and focus on efficiency is of importance. PMID- 17405466 TI - Agreement amongst examiners assessing dental fluorosis from digital photographs using the TF index. AB - OBJECTIVE: To compare the scoring of dental fluorosis by experienced examiners from digital photographs using the TF index. BASIC RESEARCH DESIGN: 120 images were selected from 703 photographs obtained during a clinical trial (Tavener et al., 2004). The selection process was stratified so that the full range of defects seen in the main study was included. The children, aged 8-10 years, were from deprived areas of Manchester, England with fluoride levels in the drinking water of less the 0.1 ppm F. The photographs of the upper and lower anterior sextants were taken after cleaning and drying the teeth. The examiners were identified by searching Medline for individuals who had previously used the TF index or had experience of scoring dental fluorosis. Of the 12 examiners identified, 10 agreed to take part. Each examiner was provided with identical CDs containing a PowerPoint presentation of the images. Twelve images were duplicated and interspersed amongst the 120 images to assess intra examiner agreement. Each examiner was also supplied with a table listing the criteria and illustrations for each of the TF index scores (Fejerskov et al., 1988). RESULTS: The prevalence of fluorosis (TF > 0) amongst the 10 examiners ranged from 43% to 70% and from 2% to 13% for the more severe scores (TF 3 or 4). Paired agreements amongst subject scores for the 10 examiners, measured using a weighted Kappa score, ranged from 0.40 to 0.71. CONCLUSION: It is concluded that although the criteria for the TF index are well defined, it is possible that examiners may interpret the criteria in different ways and conditions in which images are viewed may need to be standardised. This study may explain some of the differences in the prevalence and severity of fluorosis reported in different studies. There is a need to standardise the methods used to score dental fluorosis. PMID- 17405467 TI - Frequency and distribution patterns of sealants among 15-year-olds in Denmark in 2003. AB - AIMS: To provide information on the frequency and distribution patterns of sealants in the Public Dental Health Service for Children (PDHSC) in Denmark. A further aim was to determine whether there was a correlation between DMF-S and sealants. DESIGN: A cross sectional survey using data from 15-year-old Danes in 2003. SAMPLE AND SUBJECTS: The sample consisted of 50 randomly selected municipalities from the 204 municipalities with public clinics. Thirty-nine municipalities (78%) forwarded useful data. A total number of 3,184 15-year-olds were involved in the study. RESULTS: The mean DMF-S was 2.97 (SD = 1.40) and 42% had a DMF-S = 0. The mean number of sealants was 3.06 (SD = 1.60). Two-thirds of all participants had one or more sealed surfaces. At the individual level the correlation coefficient (rs) between sealants and DMF-S was -0.05 (p < 0.01). The mean number of sealants in one municipality was 0.26, in another 6.00. The surfaces most often sealed, were the occlusal surfaces on permanent second molars (35%), closely followed by the occlusal surfaces on permanent first molars (32%). Sealants on premolars were rare (1.5%). At the municipality level there was no significant correlation between mean DMF-S, % 15-year-olds with a DMF-S = 0 and mean number of sealants (rs = 0.02 (p = 0.90) and rs = 0.06 (p = 0.73), respectively). CONCLUSION: The data indicate a high use of sealants on molar teeth, a very large inter-municipality variation, no correlation between sealants and DMF-S. These findings suggest that there are no clear guidelines for use of sealants in the PDHSC in Denmark. PMID- 17405468 TI - Enamel defects and its relation to life course events in primary dentition of Brazilian children: a longitudinal study. AB - OBJECTIVE: To evaluate the cumulative incidence of enamel defects (ED) and its correlation with life course events such as malnutrition and pre- and postnatal infections, in a cohort of children of low socioeconomic status. METHODS: The children were registered at birth and examined from 12 to 36 months of age. At the baseline, 246 were examined and in the follow-up, 228. The teeth were examined under natural light and dried with gauze. ED was determined by the DDE index. RESULTS: Results show that gender was not significant. The cumulative incidence of ED at the last recall examination was 78.9%. The most prevalent type of defect was diffuse opacity, present on the gingival half of the buccal surface (p < 0.001). For logistic regression analysis maternal infections, intrauterine growth retardation (IUGR), malnutrition and postnatal infections were selected as predictor variables for the occurrence of enamel defects. CONCLUSIONS: The results indicate that life course events such as undernutrition and childhood infections during teeth development may be associated with enamel defects in socioeconomically underprivileged communities and may compromise the oral health related quality of life. PMID- 17405469 TI - Oral health care and status of elderly care home residents in Glasgow. AB - OBJECTIVES: To investigate variations in reported oral health care provision and unmet dental need within a sample of care homes for the elderly in Glasgow. BASIC RESEARCH DESIGN: Two-phase study of a 50% random sample of 120 care homes (n = 60). Phase 1: telephone interviews with care home managers. Phase 2: oral examinations of residents and face-to-face interviews with care staff, in a sub sample of 10 homes. CLINICAL SETTING: Care homes in Greater Glasgow. PARTICIPANTS: Residents and staff. RESULTS: Phase 1: data were collected from 58 of 60 care home managers approached (response rate: 97%). Oral assessment of residents on admission was reported for 78% of nursing homes and 24% of residential homes, and annual screening by a dentist for 85% and 76% respectively. Only 58% of nursing homes and 8% of residential homes had a formal mouth care policy. Staff training in mouth care was not universal and often carried out "in-house". All managers believed dental support was available when required. Phase 2: 288 oral examinations and 22 staff interviews took place within 10 homes selected. Examinations revealed high levels of oral disease. When interviewed, staff accepted that mouth care was within their remit but failed to document it within patient care plans. Further training on oral health and disease was requested from all levels of staff. CONCLUSIONS: Oral disease is common among elderly care home residents in Glasgow. Increased emphasis on implementation of standard oral care protocols and improved education of staff is essential if the oral health of elderly institutionalised residents is to improve. PMID- 17405470 TI - Knowledge of and attitudes toward severe acute respiratory syndrome among a cohort of dental patients in Hong Kong following a major local outbreak. AB - OBJECTIVE: To assess the knowledge of and attitudes toward severe acute respiratory syndrome (SARS) among patients attending a teaching dental hospital and private dental practices in Hong Kong during a major local SARS outbreak. METHODS: 250 dental patients were interviewed by questionnaire and 213 were interviewed by phone. RESULTS: Less than one-third (30.0%) of the 463 respondents said they were not afraid of contracting the SARS coronavirus from their dentists and did not avoid dental treatment for that reason. Nearly three-fifths (56.7%) did not worry about contracting SARS from dental treatment. Fewer than 10% of the respondents thought that dentists ran a high risk of contracting SARS. From the patients' experiences, 85.2% and 21.7% of the dentists wore face masks and face shields, respectively, when delivering dental treatment. CONCLUSIONS: The majority of patients interviewed had confidence in their dentists, their treatment environments, and the infection control measures taken, and were not worried about contracting SARS in the dental setting. This perception is an improvement from that described in an earlier study, in which more than half of the patients were concerned about contracting an infection during dental treatment and perceived that the infection control measures undertaken by the dental profession to prevent infectious diseases were not satisfactory. Patients, however, demanded better infection control measures during the SARS outbreak. PMID- 17405471 TI - Conversion tables for the Corah and Modified Dental Anxiety Scales. AB - Dental anxiety assessment can be achieved by using brief multi-item scales. Corah's Dental Anxiety Scale has been used extensively since the 1970s. However the scale has some flaws which led to the design of the Modified Dental Anxiety Scale incorporating some minor but important improvements. To enable comparison between studies who have adopted one but not both scales a conversion algorithm was required. OBJECTIVE: To produce regression equations and tables to enable researchers to convert derived scores from one scale to another. BASIC RESEARCH DESIGN: Cross-sectional survey. CLINICAL SETTING: 18 dental practices in Northern Ireland. PARTICIPANTS: Patients (n = 1,028) were invited to participate. MAIN OUTCOME MEASURES: Corah and Modified Dental Anxiety Scales. RESULTS: Twenty four patients refused (response rate 98%) providing 1,004 patients for analysis. Mean scores for both scales were close to those reported elsewhere. The correlation between the two scales was high (r = 0.89). Regression equation and summary tables presented for conversion purposes. CONCLUSIONS: More accurate estimates of scale scores can be derived with this procedure than simple prorating. PMID- 17405472 TI - An audit of paediatric dental treatments carried out under general anaesthesia in a sample of Spanish patients. AB - OBJECTIVE: To evaluate the success and failure rates of the clinical procedures carried out under general anaesthesia in disabled or medically comprised and healthy children. BASIC RESEARCH DESIGN: Retrospective study included 47 patients who received dental treatment under general anaesthesia, grouped according to whether they were disabled or medically compromised (group A, n = 16) or not (group B, n = 31), and subgrouped according to whether they were under or over 6 years of age. RESULTS: Mean duration of anaesthesia was 2 hours and 25 minutes, with a range of 1 to 4 hours. The percentage of children followed up was 87%. The procedures performed were: 105 preformed metal crowns, 142 restorations, 85 pulpotomies and 166 extractions. The success rate was 93% for preformed metal crowns, 96% for pulpotomies and 90% for restorations. CONCLUSIONS: General anaesthesia is necessary in some children, but should be complemented with a preventive programme, behavioural remodelling and a follow-up schedule to avoid having to repeat the use of general anaesthesia. PMID- 17405473 TI - The dental caries experience of 5-year-old children in Great Britain (2005/6). Surveys co-ordinated by the British Association for the study of community dentistry. AB - OBJECTIVE: This paper reports the results of standardized clinical caries examinations of 5-year-old children from across England, Wales and Scotland in 2005/6. These co-ordinated surveys are the latest in a series which seek to monitor the dental health of children and to assess the delivery of dental services. METHOD: The criteria and conventions of the British Association for the Study of Community Dentistry were used. Representative samples were drawn from participating strategic health authorities (SHAs), primary care trusts (PCTs) and health boards (HBs). Caries was diagnosed at the caries into dentine threshold using a visual method without radiography or fibre-optic transillumination. RESULTS: 239,389 five and six year-old children from across England, Wales, Scotland and the Isle of Man were examined in 2005/2006. The results again demonstrated a wide variation in disease prevalence and care strategies across Great Britain. Mean d3mft across England was 1.47 (d3t = 1.10, mt = 0.20, ft = 0.16), across Wales the corresponding values were 2.38 (d3t = 1.70, mt = 0.43, ft = 0.25) and in Scotland 2.16 (d3t = 1.45, mt = 0.51, ft = 0.20). Overall, 39.4% of children in Great Britain had evidence of caries experience in dentine (d3mft > 0, including visual dentine caries). The distribution of caries was highly skewed. Thus the mean caries experience for those with dentinal decay was 3.99, as opposed to the overall mean of 1.57. Trends over time demonstrate a small change in mean d3mft since 2003/4 when the mean was 1.62, although the mean value for those with dentine decay experience remained constant (4.00 vs 3.99). The care index has also fallen marginally from 12% to 11%. The BASCD co-ordinated NHS Epidemiology Programme will evolve in coming years as differing priorities in the frequency of inspecting particular age groups is being seen as well as a desire to measure other aspects of oral health in addition. CONCLUSION: Overall, there has been only a small overall improvement in the dental health of 5-year-old children over the last 2 years and no diminution of the level of disease in those affected for some time, although in Scotland a pattern of continuing steady progress from previously high levels is seen. While many children enjoy good oral health, sizable groups remain within the population of 5-year-old children who have a clinically significant burden of preventable dental disease. PMID- 17405474 TI - [A randomized clinical trail of 2 fast-resorbing suture materials]. AB - VICRYL rapide is a fast-resorbing synthetic suture material which is widely used in (oral) surgery. The suture wire is broken down by hydrolysis, making its removal unnecessary. The aim of this study was to compare the clinical suitability of the renewed generation VICRYL rapide with that of the original generation. 'Suitability' was defined as absence of discomfort, minimal pain for the patient, good knot security and tensile strength, predictable resorption, minimal tissue response and good wound healing. The trial comprised 120 patients who underwent an apicoectomy at the department of Oral and Maxillofacial Surgery. The patients were assigned at random to a group treated with the renewed generation VICRYL rapide (n = 60) and a control group treated with the original generation (n = 60). The trial was performed double-blind. Regarding the clinical suitability it can be concluded that the renewed generation VICRYL rapide does not yield any significant improvement over the original generation. PMID- 17405475 TI - [Post-academic dental specialities 8. Post-academic specialization in dental care for disabled]. AB - The dental treatment of people with a mental and/or physical disability often entails a variety of related problems and requires extra expertise and competencies on the part of the dentist. If treatment in an ordinary practice is not (or is no longer) possible, the patient can be referred to a centre for special care dentistry or to dental practices in institutions for the mentally disabled. In the Netherlands, a three year postgraduate programme in dentistry for the disabled was established in 2004, the purpose of which is to improve the availability and quality of dental care for the disabled. PMID- 17405476 TI - [Post-academic specialties 9. Dental care of disabled children living at home]. AB - The aim of this study was to identify possible problem areas regarding preventive selfcare and dental care for severely mentally retarded children living at home. The sample test comprised parents (n = 126) and dentists (n = 40) of children between the ages of 4 and 12 from 7 randomly selected daycare centres in The Netherlands, who were asked to fill out a questionnaire. Of the children 32% were found to receive no dental care at all, while among the children from ethnic minority groups this number exceeded 50%. Of the parents two-thirds indicated that they had encountered problems with daily dental care and 70% reported that their child's oral hygiene was in need of improvement. Of the dentists 25% were dissatisfied with the dental care they provide, and 75% felt that dental care for mentally retarded children needs improving. The results suggest that there is room for improvement in the support of parents of mentally disabled children in maintaining an adequate level of oral hygiene, and that adequate training of dental professionals in serving the needs of people with severe disabilities is wanted. PMID- 17405477 TI - [Dissertations 25 years after date 14. The development of porous polymethylmethacrylate cement as potential implant material]. AB - The development of polymethylmethacrylate bone cement with an open spongelike structure resulted as a spin-off from efforts to dampen the high exothermal peak during hardening of the cement. A cement formulation in which an aqueous gel was mixed through the organic dough showed the desired significantly reduced temperature peak and improved biocompatibility but also ingrowth of bone into the pores that had been formed by the resorbing aqueous gel in the polymerized matrix. The expectation that such a cement would provide a better fixation of total hip prostheses did not come true because of the diminished mechanical strength due to the same porosity. Better applicability of the cement was found for bony areas that were not subjected to heavy stresses such as bone defect filling and augmentation of cranio-facial and sternal deformities. In spite of successful clinical trials and positive findings after long term evaluations a commercial development of the cement was not undertaken because of a market that was estimated too small to be profitable. PMID- 17405478 TI - [Unusual location of an oral squamous cell carcinoma]. AB - A 52-year-old woman was found to have a small squamous cell carcinoma of the palatal gingiva of teeth 21 and 22. Such a location is extremely rare. Treatment consisted of a partial maxillectomy. The defect was closed by a partial denture. During the 5 year follow-up period no signs of local recurrence or metastatic spread occurred. PMID- 17405479 TI - [The path from cigarettes--with support it works better]. PMID- 17405480 TI - [Varenicline--a new drug for smoking cessation]. PMID- 17405481 TI - [Benzodiazepine withdrawal. Can the pharmacists make a contribution?]. PMID- 17405482 TI - [Management of early rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) affects 1% of the adult population and is a chronic disease lasting for many decades. RA often results, despite therapy, in progressive joint destruction, deformity and disability, causing a major economic loss for the patients and the society. Most common the onset of the disease is between the 4th and 5th decade: RA affects 3 times more women than men. Both, the diagnostic as well as the therapeutic opportunities, have greatly improved over the last years. Nevertheless, there are still deficiencies in the care of patients with early RA (ERA). Consequently, the "German Society for Rheumatology" has set up recommendations for "The management of early RA", which are introduced in this article. PMID- 17405483 TI - [Drugs from the sea]. AB - The oceans are currently rediscovered as a treasure chest for drugs, as today's researchers have more knowledge about marine organisms, as well as more sophisticated technology to specifically target pharmacologically active compounds. PMID- 17405484 TI - [Pain treatment in reduced kidney function?]. PMID- 17405485 TI - [Defensins in chronic inflammatory diseases]. PMID- 17405486 TI - [New treatment methods with chaperonin 10]. PMID- 17405487 TI - [Decreased vaccination reactions with long needles]. PMID- 17405488 TI - [Zanamivir for prevention of influenza and in children]. PMID- 17405489 TI - [How promising is decontamination treatment of MRSA carriers?]. PMID- 17405490 TI - [Spreading through sneezing, colds and allergies]. PMID- 17405491 TI - [Antiretroviral therapy: are treatment pauses advisable?]. PMID- 17405492 TI - [Synthesis plus absorption inhibitors are useful for high risk patients]. PMID- 17405493 TI - [Does obesity increase cancer risk?]. PMID- 17405494 TI - [A combination of antihypertensives with various mechanisms of action]. PMID- 17405495 TI - [Intranasal zolmitriptan acts quickly]. PMID- 17405496 TI - [Prevention of glucocorticoid induced osteoporosis with alendronate or alfacalcidol]. PMID- 17405497 TI - [Somatropin for minor growth retardation caused by glucocorticoids]. PMID- 17405498 TI - Quality care depends on quality character. PMID- 17405499 TI - Educating patients on dental care. PMID- 17405500 TI - Retreating a previous dentist's work. PMID- 17405501 TI - Principles of ethics and code of professional conduct. With official advisory opinions revised to January 2005. PMID- 17405502 TI - Communication in a time of conflict. PMID- 17405503 TI - Who is responsible for billing practices? PMID- 17405504 TI - Botox and dentistry--is it worth the risk? PMID- 17405505 TI - Should dentists guarantee treatment? PMID- 17405506 TI - Dr. Glenn Masunaca. Part of 'The Greatest Generation'. PMID- 17405507 TI - [Tryptose sulphite cycloserine agar for the recovery of Clostridium perfringens in surface waters: a study of different modes of utilization]. AB - In the recent European Drinking Water Directive, Clostridium perfringens has assumed increasing importance so as to be considered a primary contamination indicator. Therefore it emerged the necessity to make culture methods, aimed at its recovery, more specific and sensitive. In this study we have verified the ability of Tryptose Sulphite Cycloserine Agar plates (TSC Agar), prepared and stored before the use at refrigeration temperature (+4 degrees) for different times, to show typical colonies, using both, the single layer and double layer techniques. Results show that storage of the prepared medium, even for a few days, decrease the recovery of typical colonies although such negative effect is minimized by using the double layer technique. PMID- 17405508 TI - [Survey of the presence of Listeria monocytogenes in meat products sold in retail]. AB - The present study evaluates the presence of Listeria spp and particularly of L. monocytogenes in bovine, pork and poultry meats sold by retail in supermarkets and butchers in the city of Cassino. The sensibility to the antibiotics mostly used in the veterinary practice has been tested on the isolated strains. The different species of Listeria have shown a considerable variation of isolation based on the meat's typology and on the different store's provenance. Moreover our results show greater degree of contamination than the data currently available the Italian literature. In our study poultry meat is the most contaminated one. We can assert that omissions and poor caring errors in the manipulation and conservation of meat expose the customer to an even higher risk of infection. PMID- 17405509 TI - Influenza like Illness (ILI): prescribing behaviour of 83 sentry doctors in Lazio region in the period 2001-2002. Is it adherent to Italian ILI guidelines? AB - To describe different doctors' attitudes in drug prescribing in case of influenza Like-Illness during 2001-2002 influenza season in Lazio region, with regard to consumption and expenditure and its adherence to Italian Guidelines on ILI Management. Prospective study aimed to assess doctors' behaviour in prescribing in respect with the following events: (a) ILI and subsequent controls after diagnosis, (b) complications, (c) adverse events to influenza vaccine. 7,629 subjects, have been identified and only 17% presents one or more comorbidities. There are totally 7,766 cases of ILI: 23% are complicated and out of the remaining 77%, only 14% presents comorbidities. Almost all elderly people have been vaccinated. Antibiotics were prescribed to complicated cases (82%) with comorbidities (55%); 1,075 patients (12%) had second or third contact with doctors in a period longer than 7 days and about 65% of them received antibiotics in case of acute bronchitis; children received mainly ibuprofen and paracetamol for fever control; acetylsalicylic acid in children group has been delivered only in 1% of cases. Overall, doctors' attitude in prescribing is generally coherent with Italian Guidelines on ILI even though a high variability still persists. PMID- 17405510 TI - [Human intestinal parasitosis: role of Dientamoeba fragilis in human infections]. AB - The Authors report prevalences of intestinal parasitosis among home children and adults during 2002-2004, as in O&P as in acute or prolonged diarrhoea, with particular attention to the role of Dientamoeba fragilis, because often undervalued. Among 3139 subjects, 116 cases of dientamoebiasis (3.7%) and 62 of giardiasis (2.0%) were observed; not typical pathogenic protozoa were reported in 71 cases (2.3%); helminths were identified only in 8 cases (0.5%). Particularly, inside O&P group D. fragilis prevailed in 5.2% of cases (7.8% in adults and 0.5% in children) and G. duodenalis in 2.7% (3.5% and 1.3% respectively); inside acute diarrhoeas D. fragilis prevailed in 1.6% (3.9% and 0.3%) and G. duodenalis in 0.6% (1.3% and 0. 1%); inside prolonged diarrhoeas D. fragilis prevailed in 3.5 % (2.6% and 5.4%) and G. duodenalis in 3.9% (5.8% in adults and never in children). D. fragilis was more often observed among adults (6.3% of all) than among children (0.6%), like G. duodenalis (3.1% versus 0.6%). So, 107 strains of D. fragilis (92.2%) and 53 strains of G. duodenalis (85.5%) were identified in adults. D. fragilis was more frequent among females (24/39 cases, 61.5%, in the last year) while G. duodenalis was more frequent in males (13/23 cases, 56.5%). The Authors conclude underlining the importance of a permanent stain, as Giemsa stain, for a good and complete diagnosis of protozoal intestinal infections, particularly for D. fragilis. PMID- 17405511 TI - [Healthcare profiles: similarities and differences with respect to case and disease management]. AB - In Healthcare systems, to overcome the problem of the fragmented response to the medical needs of patients, new theoretical models and operative approaches have been proposed; the process-based approach is one of the solutions more appreciated. This approach has been adopted in different ways in different healthcare systems: typically the Case management, the Disease management and, in Italy, the Healthcare profiles. These three healthcare models that share in the logic of a process-based approach present similarities and dissimilarities. This article aims to describe the features of Healthcare profiles compared to Case and Disease management, using a comparison scheme which contains items capable to distinguish the different Healthcare models. PMID- 17405512 TI - [Epidemiology of femur fracture and characteristics of hospital care in Lazio]. AB - We aimed at describing the epidemiology of femur fracture in elderly hospitalized for femur fracture in Lazio (Italy), and evaluating the association between patient's and hospital characteristics on in-hospital mortality. We conducted a population- and hospital-based study (Lazio region: 5.233.233 inhabitants) among people 65+ years aged. SOURCE OF DATA: regional hospital register 2002-2003; ICD 9-CM codes for patients' selection 820 e 821. Direct standardization (rate x 1000) and logistic regression analysis (OR, 95% CI) were performed. Overall hospitalization rate in elderly was 7.5%o (10.l%o females vs. 3.9%0 males). 12.033 patients with femur fractures were enrolled in the study period; 21,6% were not treated surgically: in comparison with those who underwent surgery, they were males, residents out of Rome, older and with worst health status. In hospital mortality rate was 7,97%. In-hospital mortality determinants were: male gender (OR=0.56), older age (85+, OR=3.30), living out of Rome (OR=0.50), comorbidities (Charlson 'index 3: OR=4.44), "others and unspecified parts of femur" as site of fracture (OR=1.84), admission to a private hospital (OR=O. 79) and a surgical treatment (OR=0.20). In conclusion, this study showed the effect of selected individual characteristics on in hospital mortality and suggested a role of early surgical treatment and access to private sector. Regional hospital information systems represent useful tools to address epidemiological impact of hip fracture and its health care resources utilization. PMID- 17405513 TI - [Performance indicators: INCA (cardiovascular indicators) project]. AB - The objective of INCA project was the development and implementation of Acute Myocardial Infarction (AMI type ST elevation) process and outcome indicators for the regional cardiology units, testing the possibility of using regional healthcare information data to evaluate the quality of provided healthcare within the regional healthcare accreditation process. The project is introduced by an overview of major concepts of evaluating and managing quality of healthcare. We performed a literature review of structure, process and outcome indicators in cardiology and of accreditation standards for cardiology at national and international level. Through consensus procedures and according to international evidence based literature a set of 18 process and outcome indicators for AMI was defined. A specific procedure for data collection has been developed. Education and training of participants on procedures, quality and accreditation was achieved. Expected verifiable end-points have been achieved over a three months period of data collecting throughout 21 cardiology units, differentiated for level of complexity and location, for a total of 409 clinical observed cases of AMI. Analysis of data was followed by the diffusion of results. Successful data collection of clinical performance indicators on a regional basis was achieved. Participants have been trained to quality sciences. Results will be useful to evaluate and design implementation strategies of regional accreditation of health care services within a shared framework. Benchmarking within Regional hospital cardiology care services will be developed following self evaluation and continuous quality improvement cycle activities. PMID- 17405514 TI - [Smoking cessation and weight gain]. AB - Cigarette smoking is the single most important preventable cause of death and illness. Smoking cessation is associated with substantial health benefits, but weight gain after smoking cessation is perceived to be a barrier against quitting smoking. The aim of the study was to analyse predictors of weight gain after smoking cessation. The sample included 1067 residents, aged 18-70 years, in a health district of Rome who answered to an anonymous postal questionnaire. Among them 482 were former smokers; 398 provided lifetime histories of both body weight and smoking and were considered in the analysis. 52.5% (49.3% M; 60.5% F) reported weight gain after smoking cessation; among these 25.4% reported a weight gain > or =5 kg. The multivariate logistic regression analysis showed a direct association between female gender (OR 1.9, CI 95% 1.1-3.2), age - 45 years (45-65 years: OR 2.5, CI 95% 1.4-4.4; > 64 years OR 2.1, CI 95% 1.0-4.0), number of cigarettes per day >20/day (OR 3.8, CI 95% 1.3-11.5) and weight gain after smoking cessation. The relevance of weight gain following smoking cessation suggests that health benefits associated with smoking cessation may to some extent be negated by the detrimental effects on health of associated weight gain. Smoking cessation programmes should therefore consider incorporating follow-up support to prevent weight gain; regular measurements of body weight together with dietary indications and increase of physical activity are basic factors to implement in the intervention of smoking cessation. PMID- 17405515 TI - [Fatal home accidents in Tuscany in 2001-2002]. AB - In the last years in Italy deaths due to accidents were more than 26.000/year. One of the most frequent places in which they occurred was at home, with more than 8.000 deaths/year. Due to the lack of studies related to this field, in Tuscany a specific study of fatal home accidents has been done. The cases studied, occurred in Tuscany in 2001-2002, have been extracted from the electronic database of the Regional Mortality Registry of Tuscany. Information about the modality of the accidents was collected by administering a questionnaire to the closest available relative of the victim. A total of 284 cases were studied, the majority of whom were female, over 75 years of age, with low educational level, pensioners, often with more than three pathologies. The accidents happened mostly in the bedroom or on the stairs due to accidental falls. The dynamics of fatal accidents showed great differences if compared to the nonfatal accidents. This suggests the need of different approaches for fatal injury surveillance and for the implementing of preventive strategies. PMID- 17405516 TI - Distribution and phenotype of proliferating cells in the forebrain of adult macaque monkeys after transient global cerebral ischemia. AB - We performed transient global cerebral ischemia on adult macaque monkeys by reversibly stopping blood flow to the brain. We labeled de novo-generated cells in postischemic animals as well as in sham-operated controls by infusing the DNA synthesis indicator BrdU, and subsequently investigated the distribution and phenotype of BrdU-labeled cells in several telencephalic regions at various time points after ischemia. The ischemic insult significantly increased the number of proliferating cells in the hippocampus, SVZ, neocortex, and striatum, but had no such effect in PHR. In the olfactory bulb, ischemia did not change the proliferating cell levels in the first two postischemic weeks, but did increase these levels at long-term survival time periods. The majority of newly generated cells outside the germinative centers were of a glial phenotype, while neurons constituted only 1% of these cells. Notably, no new neurons were observed in the hippocampal CA1 sector, the region exhibiting the highest vulnerability to ischemia. Within the germinative centers, most BrdU-labeled cells were of a progenitor phenotype and a large proportion of these precursors sustained their existence in the niche for months after ischemia. Furthermore, cells with a progenitor phenotype were identified in brain parenchyma, and these might be responsible for the limited parenchymal neurogenesis as well as for the oligodendrogliogenesis and astrogliogenesis in striatum and neocortex. Our results show that ischemia differentially activates endogenous neural precursors residing in diverse locations of the adult primate CNS. A limited endogenous potential for postischemic neuronal repair exists in neocortex and striatum, but not in the hippocampus proper of the adult macaque monkey brain. The presence of putative parenchymal progenitors and of sustained progenitors in germinative centers opens novel possibilities for precursor cell recruitment to sites of injury. The molecular manipulation of this process may advance our ability to effectively apply brain progenitor cells in the treatment of human neurological diseases. PMID- 17405517 TI - [Preventive measures against health damage due to chemicals in household products]. AB - Chemicals in household products have been paid much attention as main cause of health damage on consumers, such as allergic contact dermatitis. Preventive measures against health damage due to chemicals in fabric, plastic and rubber products for household uses, are reviewed, focusing on (1) regulation and voluntary control by manufacturers, (2) incidence of health damage from household products, (3) causative product-chemical investigation, (4) case studies on skin damage and respiratory tract damage. PMID- 17405518 TI - [Hazardous food-borne fungi and present and future approaches to the mycotoxin regulations in Japan]. AB - In recent years, various food-related accidents and health scares have dissipated trust in the food industry. Health hazards resulting from food contaminated with fungi is increasing. Food contamination by fungi causes many problems, especially in Japan, which relies on foreign countries for about 60% of its food: the contamination of imported food by fungi and mycotoxins constitutes a serious problem. As the quantity of imported food increases and changes in food distribution have occurred, so too has the number and type of fungi causing food related damages; osmophilic and thermotolerant fungi, in addition to the mainstream fungi of genera Cladosporium, Pecinillium, and Aspergillus, have become a problem. Although European countries and the U.S. have recently conducted risk assessments for mycotoxins, Japan has not attained an international level in the determination of baseline values. However, in addition to risk management for Aflatoxin M1, Ochratoxin, T-2 toxin/HT-2 toxin, and Fumonisin, determination of baseline values for mycotoxins is beginning in Japan. In this review, we summarize hazardous food-borne fungi, and present and future approaches to the mycotoxin regulations in Japan. PMID- 17405519 TI - [Trends in the utilization of internet for facilitating transparent and more active stakeholder participation in the Codex process in several countries]. AB - Codex standards have become the benchmarks against which national food safety control measures and regulations are evaluated within the legal parameters of the World Trade Organization (WTO) Agreements. For this reason, high-level representation at meetings of the Codex Alimentarius Commission, it's related committees and ad hoc intergovernmental task forces continue to be a priority for many governments. Opportunities broaden for stakeholder input towards the development of government positions for all Codex work through the national Codex website. Some countries utilize a national Codex website as an effective communication tool between the national Codex Contact Point (CCP) and stakeholders. In this regard, stockholder participation in the national Codex preparation process is insufficient in Japan. One of the reasons for this could be the lack of information on Codex and the insufficient understanding of the work of Codex among Japanese stakeholders. To overcome these problems, more active and effective utilization of the Japanese Codex webpage should be considered. In this paper, we show analyses of recent trends of the information on Codex available from national Codex websites from six countries in order to identify the needs to establish a similar Japanese Codex website and possible contents of the site. The six websites of the national CCPs analyzed are regularly updated and utilized for the means of active information interchange between national CCPs and stakeholders, for example, providing basic general information on Codex, including its purpose, structure and meeting schedule, posting Codex working documents open for comment, and the Terms of Reference, key issues under discussion, and delegation reports of the previous sessions. Consequently, stakeholders interested in the paper could submit their comments to the delegate of the country whose contact details are made available on the website. This is one of the examples of active stakeholder participation. By establishing a similar communication system in Japan between the National Codex Contact Point and stakeholders, a more active stakeholder participation in the national codex process could be achieved. The web site could be used to provide information on the issues under discussion in each Committee and summaries of the Codex working documents circulated for the comments and their potential implications in Japan. PMID- 17405520 TI - [Studies for analyzing restricted ingredients such as phenylbenzoimidazole sulfonic acid]. AB - Phenylbenzoimidazol sulfonic acid (PBS) is a kind of sunscreens in cosmetics and is nominated as the restricted ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for PBS was investigated by HPLC. 1.0 g of the lotions with 1.0% PBS was exactly weighed, put into a 50-mL volumetric flask. Water was added to make exactly 50 mL and this mixture was used as the sample solution. On the other hand, 1.0 g of the creams with 1.0% PBS was exactly weighed, put into a beaker. After adding 1 mL of tetrahydrofuran and dissolving the cream, that mixture was transferred to a 50-mL volumetric flask. And then the beaker was rinsed with 1 mL of tetrahydrofuran and the rinsed solution was put together into the volumetric flask. After adding water to the volumetric flask to make exactly 50 mL, this mixture was used as the sample solution. If necessary, the mixture was filtrated with a membrane filter (0.45 microm). 5.0 mL of the sample solution was pipetted and put into a 200-mL volumetric flask. After adding water to make exactly 200 mL, 20 microL of this solution was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 mm i.d. x 250 mm), the mixture of 40 mmol/L acetic buffer (pH 3.4) and acetonitrile (3:1) with 0.8 mmol/L dodecyltrimethyl ammonium bromide and the detection wavelength of 305 nm. The working curve from 0.5 to 20.0 microg/mL showed a linear line between the concentrations of PBS and the peak areas. There was no interference of peak of PBS from the lotion and cream. PMID- 17405521 TI - [Analysis of Helindone Pink CN and Permaton Red in cheek rouge]. AB - Analytical methods for red tar colors, Helindone Pink CN (R226) and Permaton Red (R228), in cheek rouge were developed. R226 and R228 were extracted from cheek rouge with chloroform by ultrasonication. After centrifugation, the supernatant was collected for the determination of R226 and R228. Methanol was then added to the residue for the extraction of Pigment Red 57-1 (R201) and Pigment Red 57 (R202). Each R226 and R228 was separately detected by the silica-gel thin-layer chromatography using the mixture of hexane and chloroform (2:1) or (3:1), or hexane and tetrahydrofuran (THF) (2:1) as a developing solvent. For the determination of R226 and R228, the extract in chloroform was injected into the HPLC equipped with Amide colomn and UV-VIS detector (detection wavelength 535 nm and 487 nm) using the mixture of hexane and THF as mobile phase. The linearity was obtained between the peak areas and the concentrations of R226 and R228 in the range of 0.625-10 microg/ml. R201 and R202 were determined using ODS column and the mixture of acetonitrile and phosphate buffer as mobile phase. Seven cheek rouge samples were analyzed. The red tar colors listed in each cheek rouge were contained in the range of 247 to 6574 microg/g. PMID- 17405522 TI - [Studies for analyzing the prohibited ingredients such as selenium disulfide in cosmetics]. AB - Selenium disulfide is one kind of prohibited ingredients in cosmetics by the Japanese Pharmaceutical Affairs Act. We established the analytical method for selenium disulfide in cosmetics by ICP-MS. Selenium disulfide of 20 mg was put into a teflon vessel. After adding 5 ml of concentrated nitric acid and 2 ml of the shampoo into the teflon vessel, the mixture was digested with microwave-oven. After digesting, the mixture was made up to 25 ml with milliQ water and then it was filtrated through a milli-pore membrane (0.45 micro). After filtration, the solution was diluted with 7% of nitric acid and used as the test solution. The test solution of 100 microl was analyzed by ICP-MS (HP-4500, monitoring mass 82). The working curve from 10 to 1000 microg/l showed a linear line between the concentrations of selenium and the peak areas. Detection limit of selenium disulfide is 22 microg/l. There was no effect of the ingredients in the shampoo on selenium disulfide determination. PMID- 17405523 TI - [Studies for analyzing the prohibited ingredients such as disodium monofluorophosphate in cosmetics]. AB - Disodium monofluorophosphate is one kind of the prohibited ingredients in cosmetics due to the Japanese Pharmaceutical Affairs Act. We established the analytical method for disodium monofluorophosphate in cosmetics by capillary electrophoresis (CE). The tooth paste of 1 g was put into a 50-ml plastic tube. After adding 7.0 mg of disodium monofluorophosphate and 50 ml of milliQ water into the plastic tube, the mixture was ultrasonicated for 10 min. After centrifuging, the supernatant was filtrated through a milli-pore membrane (0.45 microm). After filtration, the solutionwas put into a 100-ml volumetric flask, made up to 100 ml with milliQ water and used as the test solution. The mouthwash of 1 ml and 7.0 mg of disodium monofluorophosphate were put into a 100-ml volumetric flask, made up to 100 ml with milliQ water and used as the test solution. The testing solution was analyzed by CE. The working curve from 10 to 100 microg/ml showed a linear line between the concentrations of disodium monofluorophosphate and monofluorophosphate peak areas. Detection limit of disodium monofluorophosphate is 0.3 microg/ml. There was no interference of peak of monofluorophosphate with the ingredients in the tooth paste and mouthwash. PMID- 17405524 TI - [Method for estimating influenza propagation pattern from daily variations in drug sales at pharmacies]. AB - A recently proposed method for estimating the route and speed of infectious disease propagation is applied to the data of four pharmacies located in and around Tokyo. The time lags of propagation between distant sites are calculated by the cross-correlation function of the daily variations in the amount of influenza anti-virus agents supplied at the pharmacies. A problem of which are infected earlier with influenza, adults or children, is also treated. The features of this study are the information sources of disease (pharmacies) and quantitative understanding of propagation (time lags). PMID- 17405525 TI - [Birth and death process of computer viruses]. AB - The daily variations in the number of computer viruses found attaching to e-mails and the number of accesses to the home page of a national institute in Japan are examined. The power spectral densities (PSD) of the variation in the computer viruses show a time-correlation characteristic of Markov process, but the daily access number does not (identified as white noise). Like biological viruses, the variation in the computer viruses can be described by the birth-and-death model known as a Markov process. PMID- 17405526 TI - [Progress on OECD chemicals programme (11)--SIAM 19 in Berlin, 2004]. AB - The 19th Screening Information Data Set (SIDS) Initial Assessment Meeting (SIAM 19) was held in Berlin, Germany, hosted by the Germen Federal Agency for the Environment. The initial assessment documents of four substances (CAS numbers: 92 70-6, 126-33-0,131-17-9, 7580-85-0) and one category (High Molecular Weight Phthalate Esters) at SIAM 19 were submitted by the Japanese Government with or without the International Council of Chemical Associations (ICCA) and all of them were agreed at the meeting. In this report, the documents of these substances are introduced. PMID- 17405527 TI - [Development of the databases for ADI (acceptable daily intake) and relevant information on food additives, pesticides and veterinary drugs]. AB - Databases for ADI (Acceptable Daily Intake) and relevant information on food additives, pesticides and veterinary drugs were developed. The databases we developed are easily accessible on the web, and contain ADIs, latest evaluation year, classification and use, as well as synonym and CAS registry number. The databases are designed to be easily updated by researchers as ADI and relevant information are updated or added without delay. The database for food additives has already provided from the homepage of NIHS, and the access log of the web site was 1325/month in December 2005 and 2179/month in March 2006. PMID- 17405528 TI - [Outbreaks of Salmonella in infants associated with powdered infant formula]. AB - Historically, outbreaks associated with Salmonella-contaminated milk products were recognized as early as the 1950's in the United Kingdom and Bulgaria. In the 1960's and 1970's there were also a number of outbreaks related to Salmonella in various powdered milk products. As a result, Salmonella criterion was included in the Codex Code of hygienic practice for foods for infants and children. Between 1985 and 2005 at least 6 outbreaks of salmonellosis, involving as many as 250 infants, have been associated with powdered infant formula (PIF). In 2005, in France, an outbreak affecting more than 100 infants was associated with PIF contaminated with Salmonella Agona. These reported outbreaks indicated that problems persisted. Experts from two FAO/WHO Expert Consultations, held in 2004 and 2006, concluded that intrinsic contamination of PIF with Enterobacter sakazakii and Salmonella has been a cause of infection and illness in infants, including severe disease which can lead to serious developmental sequelae and death. Most of the Salmonella outbreaks associated with PIF involved unusual Salmonella serotypes, which likely aided in the recognition of these outbreaks. In many regions of the world where Salmonella serotyping is not routinely performed, identification of geographically or temporarily diffused outbreaks could be difficult. It is therefore important to use the appropriate methodology to detect unusual strains of Salmonella that cause illnesses in infants, such as the lactose-positive strain, and to perform serotyping and/or pulsed-field gel electrophoresis (PFGE) genotyping for rapid identification of Salmonella outbreaks and to establish linkages between the illness and implicated food. PMID- 17405529 TI - [Investigation on new molecular entities of drugs approved in three regions, Japan, U.S. and EU--common and preceding approvals in current 4 years]. AB - The number of new molecular entities (NMEs) approved in 2005 was 17 in Japan, 20 in US and 18 in EU, respectively. Among 53 NMEs approved in Japan and at least one other region during 2002 to 2005, 53 NMEs had been approved in US and 25 in EU by 2005, but there were no common approvals only between Japan and EU. On the other hand, 26 NMEs were solely approved in US and EU during this period. Among 79 NMEs approved in either two or three regions, the number of preceding NMEs was 3 in Japan, 62 in U.S. and 14 in EU. PMID- 17405530 TI - [Study on environmental risk assessment of drugs: excretion forms to environment]. AB - Environmental risk assessment of human pharmaceuticals is needed to protect aquatic life from the toxic exposure because unaltered drugs and/or the metabolites are released to environment after human use. Application for new drugs shall be accompanied by an evaluation report of environmental risk assessment on basis of predicted use volume, already in US and near future in EU. In Japan, the specialists are reviewing methodology of environmental risk assessment of drugs now. To provide the basic information, we investigated excretion forms of drugs after human use for two groups of Japanese drugs; high sale products top 20 in the 2004 fiscal year and new molecular entities approved in 2004 and 2005. The assessment targets are materials produced for direct use in US, but those are active substances or active metabolites, excluding orphan drugs, vitamins, amino acids, peptides and proteins, in EU. According to EU condition, almost two thirds of 20 high sale products and one third of recently approved new molecular entities were identified to be the targets for environmental risk assessment. PMID- 17405532 TI - [Medical treatment of chronic heart failure]. AB - Throughout the last decade the therapeutic approach to heart failure has undergone a considerable change. Current treatment is not just directed towards the relief of symptoms; additional objectives are prevention and progression of heart failure. Heart failure concerns not only the heart, but also the response of the body to the diminished function of the heart. Consequently the emphasis of treatment is no longer aimed solely at promoting the excretion of salt and water. Additional objectives are limitations of neuroendocrine and cytokine activation and the reversal of the extracardiac abnormalities. This paper concerns the medical treatment of heart failure according to the recommendations of the European Society of Cardiology. PMID- 17405533 TI - [Internal medicine--update 2006]. PMID- 17405534 TI - [Anemia, this time without iron deficiency]. AB - The approach to anemia by the primary care physician is to diagnose treatable causes like nutritional deficiencies, renal insufficiency, chronic inflammatory disease or hemolysis. Measurement of reticulocytes allows a distinction of hemolytic and non-hemolytic anemia. Among hemolytic anemias it is important to recognize rare but potentially severe diseases as autoimmune hemolytic anemia, microangiopathic anemia and certain hereditary anemias. Non-hemolytic anemias can be categorized in microcytic, normocytic and macrocytic subtypes. This allows a rational stepwise approach for each category. Most anemias can be correctly classified, evaluated for underlying disease and treated by the primary care physician. PMID- 17405535 TI - [Cardiac resynchronization therapy in chronic heart failure]. PMID- 17405536 TI - [Patient satisfaction in surgery]. PMID- 17405537 TI - [Selection of artificial valve for aortic valve disease: mechanical prosthesis versus bioprosthesis]. PMID- 17405538 TI - [Aortic valve replacement with a small valve prosthesis: a review of clinical and echocardiographic studies for impact of prosthesis-patient mismatch]. AB - Aortic valve replacement (AVR) has become standard therapy for treating diseases of the aortic valve. However, the selection of a prosthetic valve is considered to be an important factor determining postoperative recovery of cardiac function and quality of life, because the use of small valve prostheses may cause residual obstruction to left ventricular outflow. The situation in which "the effective prosthetic valve area, after insertion into the patient, is less than that of a normal human valve," has been described as "prosthesis-patient mismatch (PPM)." The most commonly used measure of PPM is the valve effective orifice area indexed to body surface area (EOAI), and PPM is generally defined as EOAI < or =0.85cm(2)/m(2). In this paper, we review clinical and echocardiographic studies of small valve prostheses in the aortic position to investigate the clinical impacts of PPM on short- and long-term outcomes after AVR. Some studies have shown decreased symptom resolution, poor regression of left ventricular mass, or decreased survival with an EOAI < or =0.85cm(2)/m(2), while others have observed no adverse effects of PPM on short- and long-term results of AVR. Therefore, even in Western countries, in studies involving large numbers of patients, conclusions drawn concerning the impact of PPM differ greatly among reports. In conclusion, it is desirable to examine in detail, in many patients, whether the use of prosthetic valves with EOAI < or =0.85cm(2)/m(2) is also a risk factor for poor prognosis in Japanese patients, whose body size is in general smaller than that of Western patients. PMID- 17405539 TI - [Selection of the prosthesis for aortic valve replacement: mechanical or bio?]. AB - To clarify the effect of the valve selection on the patients' prognosis, long term outcome of the patients undergone isolated aortic valve replacement (AVR) with the mechanical or bioprosthetic valve in our institution were investigated. In protocol 1, patients of all ages with isolated AVR (n = 472, composed of 166 who received a bioprosthesic valve [group B] and 306 who received a mechanical valve [Group MI) between 1975 and 2004 were investigated. In protocol 2, primary AVR patients aged 50-65 years (n = 184, composed of 47 in group B and 137 in group M) were compared. There were no significant differences in freedom from cardiac death, freedom from bleeding, and freedom from endocarditis in Protocols 1 and 2. Significantly fewer redo AVRs were required in group M in both protocols 1 and 2. However, group M was significantly worse in terms of freedom from thromboembolism in protocol 2. As a result, group M was superior to group B in freedom from valve-related events when redo AVR included. When redo AVR was excluded, there was no difference between the two groups in freedom from valve related events in both protocols 1 and 2. There may thus be an advantage in selecting bioprosthetic valves in young adult patients who required AVR if they accepts the possibility of redo AVR in future. PMID- 17405540 TI - [Selection of prosthetic heart valves for adult patients by age]. AB - The use of bioprosthetic heart valves has dramatically increased over the last decade. In 2004, the ratio was 52% for mechanical and 48% for bioprosthetic valves in a survey by the Japanese Association for Thoracic Surgery. This increase in the use of bioprosthetic valves is related to evidence demonstrating the durability of such valves over the last 20 years. The guidelines of the Japanese Circulation Society recommend selection of prosthetic heart valves by considering the patient's age. In patients who received a mechanical valve in previous cardiac surgery, selection of another mechanical valve is inevitable. The age of 65 years is when patients are separated into groups receiving either mechanical (<64 years) or bioprosthetic (> or =65 years) valves. However, the evidence that a bioprosthetic valve is better for patients in their 60s is somewhat questionable, particularly in Japanese with a long life expectancy. Anticoagulation with warfarin in patients with mechanical valves leads to a higher incidence of hemorrhagic complications compared with bioprosthetic valves, although the incidence of thromboembolism is the same. Thus patients with contraindications to warfarin or a low risk of thromboembolism who are more than 65 years old are reasonable candidates for a bioprosthetic valve. It is also recommended that women of childbearing age receive bioprosthetic valves after being informed of the possibility and risks of reoperation. In addition to the information in the guidelines and physicians' preference for valve selection, factors such as the patient's lifestyle, wishes, cardiac function, other complications, and longevity must always be considered when selecting a valve prosthesis. PMID- 17405541 TI - [Stentless bioprosthesis for aortic valve replacement]. AB - The stentless aortic bioprosthesis was reported to reduce left ventricular mass (LVM) in the early period after implantation and have better hemodynamic performance and a larger effective orifice area (EOA) than stented valves. However, its clinical advantage over the stented valve is still unclear. We have implanted stentless bioprostheses in 61 patients and had a low operative mortality rate (1.6%). LVM index, left ventricular end-diastolic volume (LVEDV) index, left ventricular end-systolic volume (LVESV) index, and peak pressure gradient (PG) were rapidly reduced after implantation. Ten-year-freedom from cardiac-related death, thromboemolism, and structural valve deterioration rates were 91.0%, 100%, and 95.0%, respectively. Furthermore, using MRI analysis, blood velocity through stentless valves was similar to that through native aortic valves in comparison with stented valves. In conclusion, the stentless valve enables more physiologic blood flow through the valve, and thus it may be useful in patients with small annuli or those who require better hemodynamic performance to enable physical activity. PMID- 17405542 TI - [Long-term results of the Ross procedure]. AB - Aortic valve replacement with a pulmonary autograft (Ross procedure) was pioneered by Donald Ross in 1967. The advantages of the autograft valve include freedom from anticoagulation, hemolysis, and infection, optimal hemodynamic performance, and growth potential. Various materials have been used for the right ventricular outflow tract (RVOT). Currently, a cryopreserved pulmonary allograft is accepted worldwide as a useful material for this procedure. Pulmonary allografts cannot be utilized in Japan, and thus other valves and materials must be used instead. The acceptance of these procedures has been slow because of the technical demands of the operations and the inherent need for reconstruction of the RVOT, thereby placing two valves at risk. In the past 20 years, the Ross procedure has been increasingly considered for pediatric patients with a wide spectrum of congenital abnormalities. PMID- 17405543 TI - [Medium-term results of aortic valve replacement with cryopreserved homograft valves: importance of a domestic homograft valve bank]. AB - There are advantages to using aortic homografts as aortic valve replacements (AVR), particularly in patients with complex infective endocarditis. To determine the importance of a domestic homograft valve bank, our 23 surgical cases of homograft-AVR were reviewed. Since 2000, the Tissue Bank of the National Cardiovascular Center has supplied 23 aortic homograft valves for the treatment of complex aortic valve endocarditis. Fourteen of 23 patients had prosthetic valve endocarditis and 20 patients had an aortic annular abscess. The early mortality rate was 17% (4 patients), in all of whom prosthetic valve replacement had been performed previously. No recurrent endocarditis and no recurrent aortic regurgitation were noted at medium-term follow-up. An aortic homograft valve is the conduit of choice in cases of infective endocarditis and the importance of a domestic homograft valve bank should be recognized. PMID- 17405544 TI - [Looking back one year after the start of "the model project on potentially therapeutic death" from the standpoint of an applied hospital]. PMID- 17405545 TI - [What we can learn from a case of medical malpractice--an example of the importance of accurate medical record taking]. PMID- 17405546 TI - [Autonomic nervous system preserving operation for rectal cancer]. PMID- 17405547 TI - [Need of marks?]. PMID- 17405548 TI - [Delays of treatment of acute myocardial infarction with ST elevation admitted to the CCU (coronary care unit) in Alsace]. AB - OBJECTIVE: to determine the time delay from symptom onset to diagnosis and treatment of patients with persistant ST segment elevation myocardial infarction (STEMI). DESIGN: prospective observational study. METHOD: patients with symptoms onset < 24 h admitted in all 10 cardiac intensive care units in one French administrative region (Alsace). Data were recorded by doctors on duty soon after hospital admission. Patients with STEMI during hospital stay or as a complication of cardiac interventional procedure were excluded. The Kruskal-Wallis test was used to assess statistical differences between the groups (p value < 0.05). RESULT: from April to October 2004, 326 patients were admitted for STEMI. Median time between the symptoms onset and the patient's call for medical help was 60 minutes. General practitioners were the first medical contact in 41%. The time from symptoms onset to first medical intervention and from first medical intervention to coronary care unit admission were markedly shorter in patients who had directly called the Emergency Medical Services (group 15-110 patients i.e. 33% of the study population): 44 min vs 75 min otherwise (p=0,003). Median transport time was 60 min. Sixty two percent of the pts were transported by the Emergency Medical Services. The median time from symptoms onset to initiation of reperfusion therapy was 240 min. It was significantly lower in group 15 (170 min vs 286 min - p < 0,001) and for thrombolytic therapy (190 min versus 245 min for primary angioplasty, p=0,007). When thrombolysis (THL) was used, 89% of the pts could be treated during 6 hours of symptoms onset and 44% in 3 hours. For angioplasty only 4% of the pts were treated in the first 90 minutes, 9% in the 2 hours and 30% in the 3 hours of symptoms onset. If the time delay is evaluated from the 1 st medical intervention, call to reperfusion intervention was significatly shorter for THL: 91 versus 157 min, p< 0,003. Angioplasty represented 75% of reperfusion strategy in our area and THL alone only 2,7% and combine therapy 5,4%. CONCLUSION: our study documents the beneficial effect of a direct call to Emergency Medical Services. Our results also underscore the need for an effort to reduce the time to offer the best appropriate reperfusion techniques in STEMI pts: speed up the admission in the cath-lab, think about pre hospital thrombolysis followed by coronary angioplasty if necessary. PMID- 17405549 TI - [Comparison of mortality according to the revascularisation strategies and the symptom-to-management delay in ST-segment elevation myocardial infarction]. AB - The aim of this study was to compare the mortality associated to primary angioplasty and thrombolysis in patients managed for an elevated ST-segment acute coronary syndrome in less than or more than 3 hours after the onset of symptoms. We analyzed the in-hospital mortality of 846 patients (including 276 [33%] treated by primary angioplasty, 511 [60%] by thrombolysis, and 59 [7%] without revascularisation) included from October 2002 to December 2003 in a registry of patients with an elevated ST-segment acute coronary syndrome managed in less than 12 hours in Northern Alps districts. The overall in-hospital mortality was at 6.0% (51/846). For the 631 managed in <3 hours, the mortality rates were respectively at 5.0%, 4.6% and 11.1% respectively in case of primary angioplasty, thrombolysis and without revascularisation (p=0.21). For the 215 patients with pain lasting more than 3 hours, the mortality rates were at 2.7%, 10.3% and 21.7% in case of primary angioplasty, thrombolysis and no revascularisation, respectively (p=0.01). In the multivariable analysis, the OR of death in case of thrombolysis compared to primary angioplasty was at 1.65 (95% IC: 0.73 - 3.75) for patients with pain " 3 hours, and 4.98 (95% IC: 1.32-18.37) for those with pain > 3 hours. These results are in line with randomized trials conclusions and confirm the international guidelines suggesting primary angioplasty for patients with a chest pain >3 hours and either angioplasty or thrombolysis in case of chest pain < 3 hours. PMID- 17405550 TI - [Prevalence of hypertension in a disadvantaged population in Antilles: a major role for obesity?]. AB - The aim of this study was to evaluate the prevalence of hypertension and its management in a disadvantaged population, essentially composed of those receiving state subsidies and the unemployed. METHODS: The PHAPPG study : Prevalence of Hypertension in a Population Precaire Guadeloupeenne (disadvantaged population in Guadeloupe) included 2420 consecutive people considered to be 'disadvantaged' seen in the two medical examination centres in Guadeloupe between November 2001 and November 2003. RESULTS: The prevalence of obesity was considerable throughout the female population of Antilles (29%). It was recorded in 12% of men. The prevalence of hypertension in the population was 24.7% for men and 22.1% for women. Obesity was the main factor that could explain the higher prevalence of hypertension noted in Antilles, especially among 'disadvantaged' women. In effect, after correction for other risk factors and for age, this prevalence of hypertension was on average three times higher amongst the obese compared to subjects with normal weights. CONCLUSION: The prevalence of hypertension is elevated in the population of Antilles, especially among the disadvantaged. Obesity is the key factor, upon which a primary prevention policy for hypertension could be based. PMID- 17405551 TI - [Brugada syndrome and other cardiovascular abnormalities related to tricyclic antidepressants ans related drugs intoxication]. AB - The aim of this paper is to report major cardiovascular complications related to intoxication due to tricyclic antidepressants (TCA) and related drugs, especially the stabilizing membrane effect (SME) and Brugada syndrome, and identify risk factors related to cardiac toxicity. POPULATION AND METHODS: A retrospective study (35 months), including all adult patients admitted for an isolated intoxication by a TCA or a related drug. The statistical analysis of clinical signs and history as well as ECG abnormalities included parameters recorded in emergency rooms. RESULTS: 65 patients without underlying cardiovascular history were retrieved (mean age 30 + 12 years). The intoxication was intentional in all cases, and the mean estimated taken dosage (ETD) was at 749 + 436 mg. Amitriptyline was the most common drug (66%) followed by clomipramine (29%). The cardiovascular examination noted a tachycardia in 63% of cases (mean HR - 108 + 13 bpm), followed by hypotension (SBP = 80 + 40 mmHg) in 11% of cases. The ECG showed a sinus tachycardia (63%), a PR prolongation (>200 ms) in 28% of cases, a MSE (15.4%) and a type-I aspect of Brugada syndrome (15.4%). The analysis of influence of ETD on HR showed a linear relationship, with the equation HR = 82.276 + 0.039 EDT (mg) (R2=0.138, p=0.001). Analyzing the influence of type of drug on ECG abnormalities revealed a more tachycardia effect of amitryptiline vs. clomipramine (p=0.047). CONCLUSION: Tachycardia is a frequent sign of intoxication, which can be expressed by a linear function depending on ETD. The MSE is more frequently noted with amitriptyline. The clinical aspect of type-I Brugada syndrome is as frequent as the MSE but does not seem prognostic. It is unclear whether it is related to an isolated electrical aspect or it hides a genetic mutation. PMID- 17405552 TI - [Influence of age on the presumed cause of syncope in patients with the Wolff Parkinson-White syndrome]. AB - The aim of this study was to assess the causes of syncope in patients with the Wolff-Parkinson-White syndrome (WPW) and to determine whether the age of the patients was a significant factor. Forty-seven patients with a WPW, aged 11 to 72 years, underwent electrophysiological study by the oesophageal approach because of an unexplained syncope. Nineteen patients were under 20 years of age (16 +/- 3 years: group I) and 28 were over 20 years of age (40 +/- 13 years: group II). Junctional tachycardia was induced in 8 patients of group I (42%) and in 13 of group II (46%) (NS); atrial fibrillation was induced in 8 patients of group I (42%) and in 9 of group II (35%) (NS). A potentially malignant form of WPW was identified in 8 patients of group I (42%) and in 11 of group II (39%) (NS); Syncope was directly attributed to the WPW in 14 patients of group I (74%) and in 19 of group II (78%), either after identification of a serious form or induction of junctional tachycardia (6 patients of group I and 8 of group II). The rest of the syncopal episodes had various causes. There were no deaths. The authors conclude that oesophageal electrophysiological investigations enable rapid identification of a high incidence of tachycardias probably responsible for syncope in WPW. The causes of syncope and incidence of potentially severe forms of WPW were not significantly influenced by the age of the patients. PMID- 17405553 TI - [French as 2005-recommendations on the management of arterial hypertension]. AB - Self blood pressure measurements (home BP) and/or ambulatory BP measurements are recommended in mild to moderate hypertension (140/90 - 179/109 mmHg) in order to confirm sustained hypertension and identify white coat and masked hypertension. The evaluation of target organ damages (TOD) has to be integrated in cardiovascular risk estimate and taken into account in the management of hypertensive patients. Beside echocardiography, there is a place for the screening of microalbuminuria in non diabetic hypertensive patients, but these investigations should not be performed systematically. Arterial stiffness evaluation and carotid intima-media thickness quantification are not yet recommended. Cardiovascular risk (CV risk) estimate plays a pivotal role in the therapeutic decision and strategy. The cardiovascular risk grade is based on [1] the list of cardiovascular risk factors (same list AFSSAPS recommendations on dyslipidemia), [2] the presence or absence of TOD and [3] cardiovascular complications: "low", "medium", and "high" CV risk. Lifestyle modifications are recommended in all hypertensive patients. Five antihypertensive drugs are recommended for first line therapy: beta-blockers, thiazide diuretics, ACEIs, ARA II and CCBs (and fixed low dose combinations with AFSSAPS agreement for first line). In order to initiate the treatment, Evidence-based therapy (according to clinical trials conducted in different clinical situations), certain comorbid conditions (compelling indications), efficacy and side-effects in a previous experience, and the cost are the determinants of the first choice. Most hypertensive patients require more than one agent to achieve target blood pressure and for second line therapy the recommended combinations are: betablockers-diuretics, ACEIs-diuretics, ARAII-diuretics, betablockers-CCBs (DHP), ACEIs-CCBs, ARA II-CCBs and CCBs-diuretics. The delay to establish a combination therapy depend on CV risk. The BP goals are those recommended by ESH ESC 2003: BP<140/90 mmHg in all, BP<130/80 mmHg in diabetic patients and in patients with chronic renal failure. Beside lowering BP, the reduction in proteinuria <500 mg/24 h is a new goal in these high risk patients. These guidelines provide a tool for every day practice and applicability should be evaluated. PMID- 17405554 TI - [Proteomics in cardiology]. AB - Cardiovascular diseases are among the major causes of morbidity and mortality in the developed world. The molecular mechanisms responsible for dysfunction of the heart in most cardiac pathologies are still largely unknown, except that the expression of certain genes/proteins is altered. Proteomic analysis is a technology which can provide an overall understanding of changes in the level of protein expression. Especially with differential analysis, it now represents a powerful tool for interpreting all biochemical responses and their regulation. The principal technique employed is two dimensional electrophoresis (2-D gel) to separate the proteins followed by mass spectometry in order to identify them. Recently SELDI-TOF analysis, which is a complementary 2-D electrophoresis technique based on the combination of two principles, chromatography by retention on protein chips and mass spectometry, has allowed the comparison of protein profiles obtained from diverse biological samples. The publication of genome sequences for humans as well as for other species has provided evidence for the biochemical complexity, and in particular the fact that a gene does not just code for a single protein but for several, due to various alternative splicing processes, post-translational modifications etc... The combination of these various approaches has proved to be particularly interesting in the study of cardiovascular diseases with the aim of understanding the molecular mechanisms involved, providing evidence for protein interactions and identifying new biochemical factors / markers involved in the different cardiovascular pathologies. PMID- 17405555 TI - [Intracardiac echography]. AB - The role of echocardiography during non-coronary interventions is of increasing importance. They include percutaneous closure of atrial septal defects (ASD) or patent foramen ovale (PFO), percutaneous balloon mitral valvuloplasty (PMV), septal alcoholization, or interventional procedures managing arrhythmia. In all cases, echographic monitoring enables to guide the procedures, optimize and assess their results and avoid complications. This role could even increase with the development of other interventional techniques such as left auricle appendage exclusion or the percutaneous treatment of valvular diseases. This article reviews this new approach and its value in interventional cardiology. PMID- 17405556 TI - [Pulmonary embolism and right diaphragmatic paralysis: "one train can hide another"]. AB - The authors report the case of an 84 year old woman admitted for a mild pulmonary embolism associated with severe hypoxaemia. The association of a right diaphragmatic paralysis with renewed patency of a foramenovale and creation of a right-to-left shunt is probably an underestimated cause of refractory hypoxaemia. PMID- 17405557 TI - [A pitfall of transseptal catheterisation for percutaneous mitral commissurotomy: interruption of the inferior vena cava with azygos continuation]. AB - Transseptal catheterisation is a widely used technique in interventional cardiology. The authors report the case of a 37 year old woman admitted for percutaneous mitral commissurotomy of a symptomatic rheumatic mitral stenosis in whom transseptal catheterisation was impossible because of a rare congenital anomaly: interruption of the inferior vena cava with azygos vein continuation. PMID- 17405558 TI - [Myocardial infarction with normal coronary arteries: role of transesophageal echocardiography]. AB - We report the case of 74 years-old female patient hospitalized for a ST+ acute coronary syndrome with normal coronary angiography. The association of a patent foramen ovale, a deep venous thrombosis and a pulmonary embolism led us to conclude the diagnosis of paradoxical coronary embolism. This case allows us to remind different etiologies to be considered in case of myocardial infarction with normal coronary arteries, and the interest of transesophageal echocardiography for the diagnosis of its etiology. PMID- 17405559 TI - [Association of Wolff-Parkinson-White syndrome with isolated non-compaction of the left ventricle]. PMID- 17405560 TI - [The best of echocardiography in 2006]. AB - In recent years, Doppler echocardiography has undergone significant technological advances, resulting in spectacular improvements in diagnosis and quantification. This review summarises the advances made in 2006 in fields as varied as cardiac resynchronisation, cardiac failure with preserved systolic function, contrast echocardiography, 3D echocardiography and echocardiographic evaluation of valvular heart disease. Despite growing competition, but mostly complementarity with CT scan and MRI, Doppler echocardiography remains the keystone of cardiological rationale based on non-intensive investigations. PMID- 17405561 TI - [The best of valvular heart disease in 2006]. AB - In the field of valvular diseases, the year 2006 has been above all marked by the publication of new joint AHA/ACC guidelines. Besides, other publications included severe aortic stenosis, drug therapy in chronic aortic regurgitation, the indications for surgery in asymptomatic mitral regurgitation, as well as valve surgery in the elderly and mismatch severity after aortic valve replacement. In a series of 66 cases of aortic stenosis with poor LV function undergoing stress echocardiography, it has been shown that whether the absence of contractile reserve identifies a high-risk subgroup for aortic valve replacement (33% vs. 6%), this exam does not predict the functional prognosis and the post-operative LV function improvement. Hence, the absence of contractile reserve should not contra-indicate aortic valve replacement surgery, even though the prognosis is severe, since the natural prognosis is extremely severe. The question of aortic valve replacement in case of mild stenosis with the indication of CABG remains difficult. Taking into account the important mortality of redo surgery, a study was in favor of a systematic aortic valve replacement associated to CABG in patients <75 years who had a mean aortic gradient > 30 mmHg, a LVH and aortic valve calcifications. Old patients (>80 years) with severe aortic stenosis have a very severe natural prognosis, authorizing wide indications for aortic valve replacement, in the absence of important comorbidities. The use of vasodilators in severe chronic aortic regurgitation remains debated. In a randomized study comparing Nifedipine 40 mg vs. Enalapril 20 mm vs. placebo, it has been shown that the use of a vasodilator does not modify the time of valvular surgery at all. After a mean follow-up of 7 years, the rates of aortic valve replacement were identical in the 3 groups, and there wa no difference regarding the preoperative regurgitation volume, LV dimensions and mass, and LVEF. The timing for huge chronic and asymptomatic mitral regurgitation remains debated, even though the present trend is to prefer early surgery, due to the risk of sudden death. A new study evidenced the interest of clinical and echocardiographic surveillance on a prospective series of 129 consecutive asymptomatic patients with severe degenerative mitral regurgitation. The surgical indication was only based on the occurrence of symptoms, an echographic LV diameters increase, a LV dysfunction, the presence of pulmonary hypertension or atrial fibrillation. At 8 years, the overall mortality was at 91 + 3%, with a survival rate not statistically different from the expected rates; 35 patients have been operated. The 6-years and 8-years surgical indication-free survival were respectively at 65 +/- 5% and 55 +/- 6%, with a null operative mortality, and favorable data regarding post-operative survival, symptoms and LV function. The indication for surgical correction of mitral regurgitation in the elderly remains a difficult problem, due to the surgical risk. The Mayo Clinic team analyzed the evolution of 284 patients > 75 years operated for mitral regurgitation between 1980 and 1995, compared to younger subjects (65 - 74 years old, n=504, and <65 years, n=556). The age-adjusted 5-years survival was lower in the older patients, but the age adjusted observed/estimated survival ratios were similar in the different age groups. The perioperative risk, significantly higher in the older patients (p<0.001) decreased systematically for all age ranges in the most recent periods (the 1980-83 period vs. 1992-95), from 27% to 5%, 21% to 4%, and 7% to 2% respectively in patients > 75 years, 65-74 years and <65 years. An age >75 years was an independent predictive factor for the surgical mortality. These data show that, despite the surgical risk, the surgical treatment of mitral regurgitation should be extended to the oldest patients. One of the most surprising information provided in the ACC/AHA guidelines is the necessity of adding a daily dose of 75 to 100 mg of aspirin systematically to VKA in patients with mechanical prosthetic valves or in those with a valvular bioprosthesis with one or several risk factors for thrombo-embolic events (AF, history of thromboembolic event, LV dysfunction, hypercoagulable state). This major recommendation (Class 1, level of evidence B) appears surprising, since it is not actually based on definite results issued from randomized studies using these doses of aspirin. According to these guidelines, this association not only reduces the risk of thrombo-embolic events in patients with valvular prosthesis, but also decreases the mortality related to other cardiovascular causes (especially coronary artery disease), with the cost of a minimal risk augmentation for hemorrhagic complications. The importance of pejorative prognosis of mismatch (defined as an inadequacy between the aortic orifice size and patient's body surface after aortic valve replacement) has been subject to several publications this year. The substantial increase of post operative events and poor survival in patients with this mismatch imply adapted preventive strategies in terms of selection of diameters and type of aortic valvular prosthesis and associated aortic surgical techniques (such as annular enlargement or supra-annular position). PMID- 17405562 TI - [The best of thrombosis in 2006]. AB - The year 2006 was an excellent year for trials publications on antithrombotic treatments. The results of several important trials were published. Firstly, two new antithrombotic molecules, fondaparinux (anti-Xa) and bivalirudine (anti-IIa), were the object of large scale trials, in coronary artery disease. Fondaparinux, in the OASIS-5 study, was compared with the reference treatment by enoxaparin. The efficacy was equivalent in the treatment of acute coronary syndromes without ST elevation, in the reduction of ischaemic events and, above all, was associated with a reduction in severe bleeding complications and reduced mortality at 9 days and 6 months. The ISAR-REACT 2 trial demonstrated the value of abciximab, a platelet membrane glycoprotein inhibitor, in the treatment of high risk acute coronary syndromes identified by the risk in troponin levels. The ACUITY trial showed that bivalirudine was as effective as abciximab in the treatment of acute coronary syndromes in terms of prevention of ischaemic events and that it reduced the number of bleeding complications. The CHARISMA trial looked at the possible benefit of biotherapy with clopidogrel and aspirin versus monotherapy with aspirin alone in a large panel of high cardiovascular risk patients. The globally negative results should, however, be closely analysed because some subgroups (symptomatic patients) were identified, in which a possible benefit was observed in contrast to other subgroups (multiple risk factors) in which biotherapy appeared to be deleterious. The results of other less publicised trials (STEEPLE, FIDO, ESPRIT) are also commented. PMID- 17405563 TI - [The best of nuclear cardiology and MRI in 2006]. AB - In this review, original clinical studies based on scintigraphy, PET or MRI imaging are reported, regarding the main areas of cardiology including ischemia and myocardial infarction, detection of coronary artery disease in various patients subsets, cardiomyopathy and heart failure. A special topic is dedicated to tracers and technical innovations. PMID- 17405564 TI - [The best of hypertension in 2006]. AB - The TROPHY study was designed to show the feasibility of pharmacological prevention of hypertension with respect to the group of patients with "prehypertension" as defined by the JNC VII recommendations. This clinical trial compared candesartan 16 mg/day with placebo and the result at 4 years was a reduction in the relative risk of developing hypertension of 15.6%. The antihypertensive drug delayed the onset of hypertension in a mainly overweight masculine population. Staessen, Zhu and O'Brien's groups suggest measuring an index of arterial rigidity obtained from ambulatory blood pressure monitoring: the ambulatory arterial stiffness index (AASI). This is calculated as [1- slope of systolic/diastolic pressure]. The reference values for AASI vary with age from 0.50 to 0.70. The CAFE study, a spin-off of the ASCOT trial, showed that the central blood pressure decreased more than the peripheral blood pressure with the association amlodipine-perindopril as compared with atenolol and a thiazide diuretic. The capacity of an antihypertensive drug or an association of antihypertensives to decrease the central blood pressure could be a pertinent factor of evaluation to be taken into account in the interpretation of clinical trials. The study of the Italian cohort PAMELA showed a progressive increase in cardiovascular and global mortality with respect to the findings of increased blood pressure by one, two or three methods of measurement (at the office, at home, ambulatory) compared with patients declared normotensive by the same methods. This registry confirmed the implication of masked hypertension on cardiovascular prognosis and also showed that "white coat" hypertension was not completely benign. The "3 cities" study is a French epidemiological study of persons over 65 years of age. The control of the blood pressure of the treated elderly hypertensives was 57% in men and 70% in women when the cut-off was 160/95 mmHg and 31% for all patients in a cut-off level of 140/90 mmHg. Lafontan et al. are studying the mobilisation of fat induced by exercise, resistant to betablockers therapy and attributed to natriuretic peptides. This metabolic pathway could be of relevance in the metabolic syndrome and in cardiac failure. Renin inhibitors, such as aliskiren, are being developed. The outlook is the possible use of these drugs with ACE inhibitors or angiotensin II inhibitors, taking into account the risk/benefit ratio. PMID- 17405565 TI - [The best of vascular medicine in 2006]. AB - Peripheral arterial disease (PAD) remains an under-diagnosed affection, and the ankle-brachial index (ABI), a simple diagnostic method, is poorly known and seldom used, and the vascular patient's prescription list is frequently insufficient regarding results obtained in large trials with good methodology. The French ATTEST study underlines the fact that ABI is measured in less than 1 out of 3 patients with PAD. In ATTEST study, less than 10% have the triple therapy validated in PAD : antiplatelet drugs, statins and ACE-inhibitors. The international REACH registry included more than 60 000 patients suffering from atherosclerosis, including 8 000 cases with PAD. This survey evidences that in PAD patients, the annual cardiovascular complication rate is significantly higher than in patients with coronary artery disease (18 vs 13%); again PAD appears systematically under-treated when compared to CAD. These epidemiological surveys highlight the importance of screening of atherosclerotic lesions with the aim of setting an active prevention of CV complications. The new guidelines insist on the screening of PAD in patients at risk, as well as on the importance of the global management after initiating the triple therapy, independent of the CV risk factors. In a 5-year longitudinal study from an initial cohort of 2265 subjects, Aboyans et al. studied the progression of PAD by repeated measurements of ABI at the level of ankles and toes. Factors of progression for large-vessels PAD were active smoking, the total/HDL-cholesterol ratio, Lp(a) and CRP. Importantly, diabetes was not associated to the PAD progression in large vessels, but in contrast, it was the sole factor associated to the progression of PAD in small vessels. In an Austrian study published this year in the NEJM, Schillinger et al. compared balloon angioplasty versus the use of Nitinol stent for the treatment of long stenoses of the superficial femoral artery. In case of claudication, these lesions are usually treated medically, whereas surgery is required for more severe cases. The fact that stenting these long lesions of the superficial femoral artery provides benefits in terms of restenosis opens a approach for the endovascular therapy, to be confirmed by larger trials. PMID- 17405566 TI - [The best of epidemiology and cardiovascular prevention in 2006]. AB - The recent analysis of the French MONICA registries report a reduction in the incidence of fatal MI related to improvement of care whereas the overall incidence of coronary events remain stable, suggesting the need for a better primary prevention. The extensive review of the death certificates and the analysis of the death classification from the same registries indicate an under estimation of MI-related death in the national death registry. It is also confirmed that instead of 50%, approximately 80% of coronary death are explained by the four major risk factors including smoking, hypercholesterolemia, hypertension and diabetes. The international REACH registry has enrolled more than 67 000 individuals including patients with symptomatic atherothrombotic disease and patients with multiple risk factors. The analysis of baseline characteristics and of the one year FU shows a high residual risk and a lack of efficacy of secondary prevention. The existence of a symptomatic disease and the number of symptomatic localization of atherothrombosis are critical factors to predict recurrence of major vascular events Secondary analysis of the INTERHEART study provide the essence of what should any physician know about the relationship between coronary heart disease and smoking, either active or passive. Prevention with respect to this risk factor remains very insufficient. Varenicline, a new nicotinic receptor partial agonist, should help patients involved in smoking cessation program. The established detrimental effects of perioperative smoking represent a unique opportunity to promote smoking cessation in individuals scheduled for surgery. The major cardiovascular impact of second hand smoking has been recently demonstrated by the short-term effects of banning smoking in public places on the incidence of acute coronary events. The SPARCL study has demonstrated the benefit of high dose of atorvastatine to prevent recurrent acute ischemic cerebrovascular event in patients with a prior history of stroke or TIA. In the open ASTEROID study, high doses of rosuvastatine confirm the possibility of reducing the volume of coronary atheroma analyzed by IVUS. The expected benefit of glitazones to reduce the incidence of death, MI and stroke in diabetes patients with a prior history of vascular event has been confirmed in the PROactive study. Pioglitazone provided a clear reduction of recurrent vascular events in diabetes patient with a prior MI at a cost of a significant increase of the risk of heart failure. In the DREAM study, neither ramipril nor rosiglitazone have reduced the incidence of cardiovascular events significantly. The moderate benefit of the fenofibrate to prevent cardiovascular events in the FIELD study, which was carried out in diabetics mostly in primary prevention, needs to be considered after adjustment on statin use in a higher proportion of patients of the placebo group. Postprandial hyperglycaemia, analyzed by the peak of glycaemia after a load in glucose, has been confirmed as a more powerful independent predictive factor of the risk of cardiovascular event than fasting glycaemia. The systematic screening postprandial hyperglycaemia represents an interesting strategy for primary prevention which warrants further investigation. If obesity is a risk factor whose impact on morbi-mortality is well established, a French study shows that body mass index has an unfavourable influence on the cognitive functions in middle-aged men and women. PMID- 17405567 TI - [The best of interventional cardiology in 2006]. AB - The year 2006 was a landmark in interventional cardiology. Confirmations of results of large-scale trials and meta analyses, the commercialisation of new drug-elution stents, discussions about well established methods, questions about long-term outcomes of dilated patients, have made 2006 a particularly rich year in controversy, especially during its last three months. PMID- 17405568 TI - [The best of cardiac failure in 2006]. AB - This year, several studies performed were justified by the increasing importance of diastolic dysfunction, while the systolic dysfunction remains stable. Its prognosis, previously considered as benign, is similar to heart failure due to systolic dysfunction. Studies are difficult in this field and we are often satisfied by imperfect results: PEP CHF, limited by the enrollment issue, showed only a trend for a beneficial effect of ACE-inhibitors. However the study can be considered as significant if only limited to the first year of follow-up, which can be justified. The second major field of research is resynchronization therapy: echocardiographic studies keep on providing asynchrony criteria. The benefit is greater in case of less extended fibrosis, and if the heart disease is non-ischemic, and if the paced zone does not contain fibrosis. Finally, a promising study on the interest of left ventricular discharge by a long-term circulatory assistance in case of severe systolic dysfunction should be cited. PMID- 17405569 TI - [The best of arrhythmia in 2006]. AB - Even though the year 2006 did not bring any major publication reporting therapeutic trials in the field of arrhythmia, it has been marked by the publication of important international guidelines on atrial fibrillation (AF), the treatment of ventricular arrhythmia and the prevention of sudden death, as well as studies on AF ablation, the external automatic defibrillator, the risk factors of ventricular fibrillation in the acute phase of myocardial infarction, and genetic aspects of the arrhythmogenic right ventricular dysplasia. PMID- 17405570 TI - [The best of cardiac pacing in 2006]. AB - In the field of cardiac pacing, the past year has not been marked by the publication of decisive scientific works, but several important publications have continued to witness the dynamism of this discipline. Selecting the significant facts in 2006 was not easy. Yet, in the following lines, we have integrated them in the dynamic of this speciality. The validation of the concept of resynchronization is now definitively accepted, and its actual place in the strategy of treatment of heart failure is clearer. With the issue of the systematic adjunction of an anti-tachycardia function, we reach the meeting point between cardiac pacing, arrhythmology and heart failure. PMID- 17405571 TI - [The best of cardiac rehabilitation in 2006]. AB - The meta-analysis showing the benefits of physical training revisited: Taylor examined only the cardiac rehabilitation trials of exercise intervention alone (versus usual care) and demonstrated that cardiac mortality is 28 % reduced and exercise appears to have an independent mortality benefit. An economic evaluation of cardiac rehabilitation: a systematic review of 15 economic evaluations. Evidence to support the cost-effectiveness of supervised cardiac rehabilitation compared with usual care in myocardial infarction and heart failure was identified. But further well-designed trials are required. Pronostic value of some variables determined by exercise testing entering cardiac rehabilitation and after physical training. A beneficial effect of physical training versus usual care on BNP and neurohormones in patients with chronic heart disease. Patients on beta blockers after myocardial infarction: determination of a more accurate training heart frequency derived from the classical Karvonen's formula. The combination of trimetazidine with exercise training provides greater improvements in functional capacity, left ventricular function and the endothelium-dependent relaxation of the brachial artery than exercise training alone in patients with ischaemic cardiomyopathy referred for cardiac rehabilitation. Guidelines for resistance exercise after cardiac event: a new paradigm less restrictive, safe and efficient to accelerate patients' return to daily activities. Recommendations for participation in leisure-time physical activity and competitive sports for patients with ischaemic heart disease: the result of consensus among experts from the ESC study group of sports cardiology. PMID- 17405572 TI - [The best of fundamental research in 2006]. AB - The Cardiovascular Research Study Group attempts to organise round tables during its annual meetings based around themes that will allow clinical and fundamental researchers, especially the younger ones, to present their work. For example, certain round table subjects at the Spring cardiology meeting, such as 'Genetics: monogenic and polygenic diseases', 'Cellular therapy', and 'Ionic homoeostasis and cardiac arrhythmias', allowed several French laboratories to publish in some of the best journals such as Nature Genetics, Nature Methods, Circulation, and Circulation Research. During the Toulouse congress, eight poster prizes were awarded to young nonstatutory researchers under the age of 32 years. For the American Heart Association congress, eight travel grants were allocated to students whose abstracts had been accepted, paving the way for future publications from young French researchers in the best journals. PMID- 17405573 TI - [The best of clinical pharmacology in 2006]. AB - The clinical pharmacological and therapeutic working group was particularly impressed by twelve recent publications relative to its various themes of interest. Two studies were made of the prognostic impact of non-observance of treatment which seems to be associated with an extra-mortality even when the treatment is placebo: the probable explanation is that the non-observance of drug therapy is also associated with the non observance of dietary/life style measures and with cognitive dysfunction associated with more severe cardiac disease. A recent study on n-acetyl-cysteine has rekindled the debate on this substance for preventing nephrotoxicity of radiological contrast used during angioplasty in high risk patients. The risks of AINS drug therapy has been reassessed. The increased risk of myocardial infarction is confirmed with celecoxib but not with "classical" AINS drugs if not prescribed for more than one year and without aspirin. With respect to lipid-lowering drugs, should statins be prescribed to attain a target value of LDL-cholesterol or to attain a given reduction in LDL cholesterol? The death knell of fibrates has more or less been rung by the results of the FIELD study and the real value of OMEGA-3 drugs should be reassessed by good quality prospective studies. In the domain of hypertension, the recent arrival of aliskiren, the first of the antirenin drugs, is noteworthy although its role in the therapeutic strategy, remains to be defined. Finally, a comment is made on the results of the TROPHY study which suggest value in the possible prevention of hypertension with angiotensin II inhibitors in patients at risk of developing hypertension. PMID- 17405574 TI - [The best of congenital heart disease in 2006]. AB - As in previous years, the end of 2005 and the year 2006 were very fruitful in publications on congenital heart disease in children and adults. Interventional cardiology is still the object of randomised and non-randomised trials in adults and children. The closure of the foramen ovale is still popular in the context of a cerebrovascular accident whether or not associated with migraine. Several articles have studied the percutaneous closure of atrial septal defects: different devices, different indications, complications, comparison with surgical closure. Some ventricular septal defects can be occluded by the percutaneous approach. Another subject of interest has been theuse of the cutting balloon in peripheral pulmonary stenosis and the implantation of stents in obstruction of conduits between the right ventricle and pulmonary artery. The problem of arrhythmias in congenital heart disease is of increasing interest: the implantable automatic defibrillator, resynchronisation, thromboembolic complications, atrial flutter in babies. The outcome of congenital heart disease is a subject of great interest to paediatric cardiologists who follow up patients operated for transposition of the great arteries, tetralogy of Fallot or complex congenital heart disease by Fontan's procedure in their infancy. The results in women operated in their childhood are of particular interest: fertility and pregnancy in different cardiac lesions, complications during and after pregnancy. PMID- 17405575 TI - Use of drug-eluting stents decreasing amid safety concerns. PMID- 17405576 TI - The give or take of retiree health care benefits. PMID- 17405577 TI - Assessment of outcomes and parental effect on Quality-of-Life endpoints in the management of atopic dermatitis. AB - The objective of this study was to assess the consequences of atopic dermatitis/ eczema on two areas: (1) the quality of life of parents/caregivers and (2) resource utilization from two large group practices. Data from 414 patients with atopic dermatitis, aged two to 12 years, were collected between January 2001 and December 2003. Parents/caregivers completed the Parent's Index of Quality of Life Atopic Dermatitis (PIQoL-AD). One-way analysis of variance and analysis of covariance models determined statistical significance. Pairwise significance testing was performed to determine statistical differences (P < .05). Mean patient age was 6.7 years and 55% of patients were males; mild and moderate atopic dermatitis was present in 82% and 13% of patients, respectively. Mean PIQoL-AD scores worsened (5.9 +/- 5.4 vs. 3.0 +/- 3.6, P < .001) for caregivers whose child had disease flares versus those without disease flares. Patients with atopic dermatitis incurred an additional 1.8 unscheduled visits annually at a cost of $93.54 per patient. It was determined that atopic dermatitis may have considerable quality-of-life and financial consequences to both family and community. PMID- 17405578 TI - Failed promises: the demise of the original TennCare vision. AB - Managed care was the intended foundation of Tennessee's Medicaid-replacement program (TennCare) when it was implemented in 1994. By the Summer of 2005, however, when the mandated disenrollment of approximately 200,000 adult TennCare beneficiaries commenced, it was obvious that the promise of managed care had not been realized. The program collapsed under the weight of mismanagement and unrealistic expectations. Whether the demise of the original TennCare vision was the result of fundamental design flaws in the concept of managed care or poor execution of that concept are questions that remain unanswered, subsumed in the aftermath of pervasive failures and disillusionment. This paper identifies lessons learned from the TennCare program. PMID- 17405579 TI - Managing opioid dependence effectively in a managed care environment. PMID- 17405581 TI - Educating patients on hype versus science. PMID- 17405580 TI - Bisphosphonate use in the treatment of Paget's disease of the bone: analysis of claims in a large database. AB - This study examines the use of bisphosphonates for treating Paget's disease of the bone through an analysis of drug, dose, and therapy-duration data. Data were obtained from a national, multi-managed care plan claims database (based on claims filed from 1996-2004). Patient eligibility criteria included at least one claim with Paget's disease International Classification of Diseases, Ninth Revision code 731.0, at least one bisphosphonate pharmacy claim, at least six months' continuous enrollment after initiating bisphosphonate therapy (index date), and no more than one claim for osteoporosis. The proportion of patients with bisphosphonate therapy extending beyond the initial recommended treatment regimen (incremental drug use) and associated costs were calculated for each drug. Of 433 patients with Paget's disease receiving bisphosphonate treatment (mean age, 65.0 yr; 64.3% women), 53.1% were prescribed alendronate at the index date; 34.9% and 7.4%, were prescribed risedronate and etidronate, respectively. More than 40% of patients with Paget's disease receiving at least one bisphosphonate were prescribed bisphosphonates beyond the recommended regimen duration at substantial incremental costs. PMID- 17405582 TI - The prevention and treatment of pertussis. AB - Since 2004, more than 25,000 cases of pertussis have been reported in the United States each year. Symptoms in adults range from mild cough to severe persistent cough with posttussive emesis. The characteristic paroxysmal cough (whoop) may be absent, and the diagnosis is often missed, especially when the cough is mild. The most useful diagnostic test to confirm pertussis is a polymerase chain reaction assay of a nasopharyngeal swab sample. Patients are infectious for three weeks from the cough onset. Antibiotic treatment, preferably with macrolide antibiotics, is indicated during this time for pertussis treatment and prophylaxis. In 2005, two new acellular pertussis vaccines were licensed: one (Boostrix) for patients aged 10 to 18 years and another (Adacel) for patients aged 11 to 64 years. PMID- 17405583 TI - Pre-school viral wheeze in primary care. AB - Each year in the U.K., around a quarter of pre-school-aged children (i.e., those aged 1-5 years) present to a GP with wheeze. In most of these children, wheeze is episodic and associated with viral infection of the respiratory tract. Here we review diagnosis, assessment, treatment options and preventive strategies in pre school children with viral wheeze. PMID- 17405584 TI - Buprenorphine for opioid dependence. AB - In England and Wales, estimates suggest around 250,000 people have serious drug problems such as dependency, that cause considerable harm to themselves and others, and give rise to high social and economic costs. The number of people receiving specialist treatment for drug problems has increased greatly in recent years. Many people dependent on opioids will require opioid substitution treatment at some time. This may involve long-term maintenance to reduce use of illicit drugs, and/or short-term detoxification to stop such use completely. Standard management involves methadone maintenance therapy. Buprenorphine (Subutex-Schering-Plough) is also licensed for the management of patients with opioid dependence. Here we review the evidence for the use of buprenorphine compared with methadone for opioid dependence. PMID- 17405585 TI - What role does the case manager play in a litigated case? PMID- 17405586 TI - Self-paced exercise program for office workers: impact on productivity and health outcomes. AB - The impact of a self-paced exercise program on productivity and health outcomes of 32 adult workers in a large federal office complex was investigated during 3 months. Walking was the sole form of exercise. The first month, during which no walking occurred, was the control period. The second and third months were the experimental period. Participants were divided into three levels based on initial weight and self-determined walking distance goals. Productivity (using the Endicott Work Productivity Scale), walking distance (using a pedometer), and health outcomes (blood pressure, weight, pulse rate, and body fat percentage) were measured weekly. Results from this study, based on a paired t test analysis, suggest that although the self-paced exercise program had no impact on productivity, it lowered blood pressure and promoted weight loss. Further study using a larger sample and a controlled experimental design is recommended to provide conclusive evidence. PMID- 17405587 TI - Combined diet and exercise intervention in the workplace: effect on cardiovascular disease risk factors. AB - This study assessed the effectiveness of a 12-week pilot employee wellness program in reducing risk factors for coronary heart disease. Fifty university employees with at least one cardiovascular disease risk factor participated in the program. Interventions focused on diet, exercise, and monthly workshops. Pre- and post-intervention measurements included weight, body composition, blood pressure, total cholesterol, low-density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol, total cholesterol/HDL cholesterol ratio, triglycerides, and blood sugar. Twenty-five employees had post-intervention measurements. A survey was administered to assess adherence. The correlation between adherence and improvement in cardiovascular disease risk factors was also tested. Significant differences were observed between pre- and post-intervention measurements of total cholesterol, LDL cholesterol, total cholesterol/HDL cholesterol ratio, triglycerides, and weight. A significant correlation existed between self-reported level of participation in the diet aspect of the program and improvement in LDL levels. This multi-component, 12-week pilot employee wellness program was effective in reducing cardiovascular disease risk. PMID- 17405588 TI - Occupational safety and health objectives of Healthy People 2010: a systematic approach for occupational health nurses--Part II. AB - Occupational safety and health objectives 20.6 through 20.11 focus on reducing work-related assaults, lead exposure, skin diseases and disorders, needlestick injuries, and work-related, noise-induced hearing loss and promoting worksite stress reduction programs. Using the intervention strategies provided, occupational health nurses can play a key role in reducing workplace-related injury, disease, disability, and death. variety of resources pertaining to occupational health and safety from the federal, national, health care, nursing, and environmental realms can assist occupational health nurses in developing and implementing programs appropriate for their workplaces. Through the Healthy People 2010 occupational health and safety objectives, occupational health nurses have the opportunity to develop and implement workplace policies and programs promoting not only a safe and healthy work environment but also improved health and disease prevention. Occupational health nurses can implement strategies to increase quality and years of life and eliminate health disparities in the American work force. PMID- 17405589 TI - Email--from "to" to "send". AB - Occupational health nurses' ability to use email efficiently and effectively enhances their professionalism, responsiveness, and overall communications. Poorly written emails can damage the reputations of nurses and their organizations. Knowing when not to communicate by email is as important as being able to write effective emails. Many safeguards can be employed to prevent or minimize confusion and strengthen email communications. Names and addresses must be checked, appropriate subject lines used, and humor applied conscientiously. All emails should be proofread prior to sending, and replies must be carefully considered. Simple, colorless email backgrounds make downloading quicker. Sharp, clear fonts are easier on the eyes. Direct and defined requests and confirmations augment communication. Once "Send" is pressed, the email is out of the sender's control. Lack of network security, forwarding by others, and potential confidentiality breaches can lead to legal or public relations controversies. Email is an important business tool. Occupational health nurses must be competent in the skills of effective email communication, as it is an integral part of business and health care. PMID- 17405590 TI - Handwashing: back to the basics. PMID- 17405591 TI - Outcome of infrapopliteal bypass in the elderly. AB - OBJECTIVE: To review the published papers reporting on the use of infrapopliteal bypass in elderly patients (aged 70 years or more) in the treatment of critical limb ischaemia. METHODS: A MEDLINE (1966-2006) search for articles relating to the topic. Eight single centre studies were identified and in addition one population based study from Finland. RESULTS: Perioperative mortality rate after infrapopliteal bypass surgery among elderly patients ranged from 2% to 12%. Patency and limb salvage rates were comparable to those reported in literature for younger patients. Data on survival rates were inconsistent. Several authors propose an active reconstruction policy in the elderly provided that a successful bypass procedure is likely to save the leg and improve the quality of life of these patients by maintaining ambulatory function and independent living at least for a year. CONCLUSIONS: Despite a relatively short life expectancy, after successful revascularisation, the majority of elderly patients can be expected to spend their remaining years ambulatory and at home. The decision whether to reconstruct or amputate is always patient bound, taking into consideration different risk factors. Age alone should, however, seldom be a reason to decline infrapopliteal bypass. PMID- 17405592 TI - Management of the 'normal' appendix during surgery for right fossa syndrome: proposed clinical algorithm. AB - The management of the normal appearing appendix during laparoscopy for right fossa syndrome remains a matter of debate when no other cause is found. Recent data suggest that the appendix modulates both the normal brain-gut axis and the pathophysiology of inflammatory bowel disease by neuro-immunological pathways. Recurrence of symptoms caused by acute appendicitis is uncommon when the appendix is left in place. The decision to remove a normal appendix should therefore not be taken lightly, but after consideration of the patient's age, medical history and expectations as well as the timing and specificity of the presenting symptoms. An overview of the relevant literature is provided with an algorithm to aid in clinical decision making. PMID- 17405593 TI - Infections of aortobifemoral prostheses at level of the Scarpa's triangle--place of local treatments. PMID- 17405594 TI - Literature review of cranial nerve injuries during carotid endarterectomy. AB - OBJECTIVE: In the recent prospective randomised trials on carotid endarterectomy (CEA), the incidence of cranial nerve injuries (CNI) are reported to be higher than in previously published studies. The objective of this study is to review the incidence of post CEA cranial nerve injury and to discover whether it has changed in the last 25 years after many innovations in vascular surgery. METHODS: Generic terms including carotid endarterectomy, cranial nerve injuries, post CEA complications and cranial nerve deficit after neck surgery were used to search a variety of electronic databases. Based on selection criteria, decisions regarding inclusion and exclusion of primary studies were made. The incidence of CNI before and after 1995 was compared. RESULTS: We found 31 eligible studies from the literature. Patients who underwent CEA through any approach were included in the study. All patients had cranial nerves examined both before and after surgery. The total number of patients who had CEA before 1995 was 3521 with 10.6% CNI (352 patients) and after 1995, 7324 patients underwent CEA with 8.3% CNI (614 patients). Cranial nerves XII, X and VII were most commonly involved (rarely IX and XI). Statistical analysis showed that the incidence of CNI has decreased (X(2) = 5.89 + 0.74 = 6.63 => p-value = 0.0100). CONCLUSION: CNI is still a significant postoperative complication of carotid endarterectomy. Despite increasing use of CEA, the incidence of CNI has decreased probably because of increased awareness of the possibility of cranial nerve damage. PMID- 17405595 TI - Necrotizing fasciitis: case report and review of literature. AB - We report a case of necrotizing fasciitis of the lower limb. This medico-surgical emergency is a life-threatening invasive soft-tissue infection which primarily involves the fascia superficialis and rapidly extends along subcutaneous tissue with relative sparing of skin and underlying muscles. Clinical presentation includes fever, signs of systemic toxicity and pain out of proportion to clinical findings. Paucity of cutaneous findings early in the course of the disease makes diagnosis challenging. The confirmation of the diagnosis is often made after surgical debridement. Delay in diagnosis and/or treatment correlates with poor outcome, leading to sepsis and/or multiple organ failure. Radiologic studies including plain radiographs, CT-scan or MRI may help to diagnose necrotizing fasciitis. Prompt surgical debridement, intravenous antibiotics, fluids and electrolytes management and analgesia are mainstays of the therapy. Adjuvant treatments like clindamycin, hyperbaric oxygen therapy and intravenous immunoglobulins are discussed. PMID- 17405596 TI - Use of vacuum assisted closure in vascular graft infection confined to the groin. AB - PURPOSE: In this report we share our experience with the use of the VAC system as a less invasive means of graft preservation and an effective alternative to routine muscle flap closure, in patients with groin wound healing complications following lower limb vascular procedures. We also review the English literature regarding the use of VAC therapy on infected groin wounds when the infection affects the prosthesis. PATIENTS AND METHODS: eight patients treated with delayed healing of a groin incision following a femoral artery surgery. In six cases local exploration or CT examination showed evidence of graft involvement (Szilagyi grade III). RESULTS: Mean duration of VAC use was 21.5 days (range, 10 to 45). The wounds were filled with granulation tissue by day 10 with no purulent inflammatory exudates. At the end of VAC therapy, final closure was easily achieved by either healing by secondary intention or delayed primary closure. No patient required use of muscle flaps. There were no reinfections at 1 to 28 month follow-up (mean, 17.2 months/ one case lost to follow-up). CONCLUSION: Our initial experience with VAC therapy to treat non healing groin wounds following vascular reconstructions is very promising. Negative pressure therapy resulted in control lymph leakage, achieving healing and managing infection. PMID- 17405597 TI - Evaluation of the sentinel node biopsy in colorectal carcinoma including the results of immunohistochemical examinations. AB - AIM: To evaluate the method and results of sentinel node biopsy including immunohistochemical examinations in resectable colorectal cancer. MATERIAL AND METHODS: From April 2004 to April 2005, sentinel node biopsy was carried out with the dye method in 27 patients operated on for colorectal cancer. The standard examination of sentinel nodes consisted in the evaluation of individual H&E-dyed specimens from bisection of the node. The negative sentinel nodes were examined with the use of immunohistochemistry against cytokeratins AE1/AE3. FINDINGS: The sentinel node was identified in 25 patients (92.6%). In a routine histopathological examination it included metastases in 3 cases. The sentinel node was clean in one patient whereas other regional nodes resected "en bloc" with the tumour included metastases. The sensitivity of the method was 75%, and the number of falsely negative results was 25%. Metastases (micrometastases) in the sentinel node were found in 2 other patients (8%) in the immunohistochemical examination. This examination did not change the results of the analysis in the patient with positive non-sentinel nodes and with the negative sentinel node in H&E dyeing. Nevertheless, the sensitivity of the method rose to 83.3% and the number of falsely negative results dropped to 16.7%. CONCLUSIONS: 1. The sentinel node biopsy using the dye method is a safe and relatively easy technique showing a high success rate (92.6%). 2. Low sensitivity of the method, as reflected in the literature, may result from lack of extended histopathological examinations performed on the sentinel node (e.g. immunohistochemistry). 3. Further research is necessary to determine the role and importance of the sentinel node biopsy in colorectal cancer. PMID- 17405598 TI - Early and late term management in caustic ingestion in children: a 16-year experience. AB - Accidental corrosive ingestion is one of the common problems causing serious esophageal strictures in children. The acute phase treatment has a great effect on stricture development. In this study we aim to present our experience in the management of caustic ingestion, particularly during the acute phase. From January 1990 to January 2005, 296 children were admitted to our clinic with caustic ingestion. Ninety-one patients who received dilatation treatment due to esophageal strictures constituted the present study group. Forty-three of them were admitted to our centre immediately after caustic ingestion (Group A) whereas 48 of them received some kind of treatment in other hospitals and were referred us with the diagnosis of stricture 6 to 12 weeks after ingestion (Group B). In the acute phase, the patients were given nothing orally until esophagoscopy was performed in the first 24-48 hours. The patients with grades 2b and 3 lesions underwent a week of esophageal rest by using a nasogastric tube. IV fluids and broad-spectrum antibiotics with a single-dose steroid were given. IV ranitidine was also added to the medical treatment. If there were stricture formations on barium meal after 3 weeks, these patients underwent esophageal dilatation programmes. The response rates to dilatation treatment were higher in group A. In addition, increased perforation rates were observed in group B. Sixty per cent of patients in group A but none of the patients in group B have recovered in the first year. In conclusion, after caustic ingestion, esophageal rest combined with supporting treatment seems to provide a good success rate with respect to prevention of stricture development and other troublesome complications. PMID- 17405599 TI - Surgery of the carotid artery: local anaesthesia versus general anaesthesia. AB - OBJECTIVE: Carotid artery surgery is safely and commonly performed under general, regional or local anaesthesia. The aim of the study was to compare local and general anaesthesia in carotid artery surgery in order to establish whether differences exist in terms of peri-operative results, use of intra-operative shunts and costs. METHODS: We retrospectively reviewed the data on 426 patients who underwent carotid endarterectomy with either local or general anaesthesia at our institution over a four-year period. All available clinical, pathologic and postoperative data were reviewed and analyzed for postoperative results. Surgical indications, outcome, operative techniques, and complications were compared. RESULTS: A total of 306 carotid endarterectomy operations under local, and 127 under general anaesthesia were performed and analyzed. Groups were similar in terms of age, sex and pre-operative risk factor distribution. The local anaesthesia group was associated with a lower incidence of shunt placement and operative time when compared to the general anaesthesia group. Postoperative intensive care unit requirement, hospital stay and costs were also lower with local anaesthesia. Significant difference in neurological complications and mortality rate was not observed between the two groups. CONCLUSION: Carotid endarterectomy performed under local or general anaesthesia is associated with low morbidity and mortality rates. Local anaesthesia enables the surgeon to assess the neurological status during the procedure. It is also associated with decreased shunt usage, decreased operative time and, in high risk patients, lower intensive care unit requirement and hospital stay. PMID- 17405600 TI - Liposarcoma of the spermatic cord: report of 2 cases. AB - These case reports describe the rare condition of liposarcoma of the funiculus spermaticus. It is treated by radical surgical resection. The diagnosis is usually made postoperatively. The role of adjuvant radio- or chemotherapy is uncertain. PMID- 17405601 TI - Ileo-ileal intussusception secondary to a lipoma: a literature review. AB - Intussusception is rare in adults and it can be a challenge to diagnose on admission. Non-specific and variable signs and symptoms, frequently only occurring episodically, may cause a considerable delay before treatment. However, in 90% a predisposing organic cause can be found in adults. A case is presented of small bowel intussusception secondary to a lipoma in a 54-year-old man in whom diagnosis was suggested by CT-scan. The patient was treated with a laparoscopic assisted reduction and extracorporeal partial small bowel resection, followed by a latero-lateral anastomosis. This case serves as the basis of a review of small bowel intussusception in adults secondary to lipomas. It focuses on the rarity of the disease, but stresses the need for early referral and investigation in middle aged patients with recurrent abdominal symptoms. PMID- 17405602 TI - Intestinal obstruction due to meckel's diverticulum: a rare presentation. AB - Meckel's diverticulum occurs in about 1-3% of general population. The majority of them are asymptomatic and incidentally found at laparotomy. The most common complication due to Meckel's diverticulum in adults is intestinal obstruction. The frequency of symptoms decreases with age. Enteroliths are rarely formed in a Meckel's diverticulum and are known to cause intestinal obstruction. These should be considered in the differential diagnosis of radioopaque shadows in the plain abdominal films. We describe a rare presentation of Meckel's diverticulum in an elderly woman. PMID- 17405603 TI - Conservative treatment of a cervical thoracic duct cyst: a case report. AB - Cysts of the thoracic duct are uncommon entities that can occur in the abdominal segment, the cisterna chyli, the thoracic segment and the cervical segment of the thoracic duct. The rarest presentation is in the cervical segment, with only seventeen cases reported in English literature. The diagnosis can be made by puncture and with the use of computed tomography or ultrasonography. The cystic fluid always contains an excess of T-lymphocytes and triglycerides. Except for two cases, all reported cervical thoracic duct cysts were surgically treated by excision and ligation of the lymphatics connected to the cyst. We present a case of a successful non-operative treatment of a cervical thoracic duct cyst that was resolved by repeated aspiration and dietary changes only. PMID- 17405604 TI - Tuberculosis of the thyroid gland: a case report. AB - Thyroid tuberculosis is a very rare condition even if the incidence of extrapulmonary forms of tuberculosis has increased. We report the case of a 56 year old female patient with tuberculosis of the thyroid gland and tubercular lymphadenitis of the neck mimicking thyroid malignancy. The diagnosis was established on histological examination after surgery in August 2002. Total thyroidectomy and central neck dissection were performed for very hard euthyroid multinodular goiter and paratracheal bilateral lymphadenopathy. There were no evidence of tubercular involvement of the other organs. The patient underwent combination treatment with antitubercular drugs for 6 months. During the three years follow-up period there was no evidence of disease recurrence. PMID- 17405605 TI - Liposarcoma of the thyroid gland: a case report. AB - Primary liposarcoma of the thyroid gland is exceedingly rare with only five previous reports in the literature. We report a case of a 40-year-old male patient with nodular goitre with the suspicion of malignancy in Fine Needle Aspiration Biopsy. Total thyroidectomy was performed. The histopathological examination revealed a well-differentiated thyroid liposarcoma. The patient recovered uneventfully. Postoperatively, radiotherapy was given to the neck region. During a 2-year follow-up period, no recurrence of the disease occured. PMID- 17405606 TI - Primary ovarian cancer presenting with axillary lymph node metastases: a report of two cases. AB - BACKGROUND: Axillary lymph node metastasis of primary ovarian cancer is rare. CASE I: A 74-year-old woman presented with a 2 x 2 cm hard, mobile mass in the right axilla. She had a history of stage IIIA epithelial ovarian cancer which was diagnosed and treated four years previously. A right lateral wall involvement of the rectum was detected in abdominal tomography. A right axillary lymph node dissection and low anterior resection of the rectum were performed. Histopathologic examination showed ovarian epithelial serous papillary adenocarcinoma metastases to axillary lymph node and the rectum. CASE 2: A 38 year-old woman presented with a 3 x 2 cm hard, mobile mass in the right axilla. She was treated surgically and by systemic chemotherapy with a diagnosis of stage IIIA epithelial ovarian cancer two years previously. A trucut biopsy was taken from the enlarged axillary lymph node, and histopathological examination revealed metastases of primary ovarian cancer. Complete axillary lymph node dissection was performed and metastases of ovarian papillary adenocarcinoma were found in 11 of the 30 lymph nodes. CONCLUSION: Supradiaphragmatic lymph node involvement of primary ovarian cancer is very rare. We report here two cases presenting with axillary metastases of ovarian cancer. PMID- 17405607 TI - Anterior thoracic wall traumatic lymphocele: a case report. AB - Traumatic lymphoceles are usually the result of surgical injuries or blunt trauma. Although usually described in the pelvis after radical pelvic node dissection or kidney transplantation, traumatic lymphoceles are rare. Diagnosis is based on CT scan and confirmed by fine needle aspiration with biochemical analysis. Treatment modalities are not standardized and different techniques have been described. We report herein a case of anterior thoracic wall lymphocele due to blunt trauma treated in our department. PMID- 17405608 TI - Temporary aorto-aortal bypass used during extended pneumonectomy for lung cancer. AB - Involvement of the aortic arch, or the descending aorta, is not uncommon in left sided lung tumours. The direct invasion of the aortic wall is generally considered a contraindication in lung resection. However, there are a limited number of reports of full thickness resections of the aorta during lung cancer surgery. They may be accomplished as a partial resection using a patch, or as a local tubular resection with reconstruction by a graft. In order to prevent ischaemia of the spinal cord, a cardiopulmonary bypass is usually recommended. The authors present a case report of a full thickness tubular resection of the descending aorta during pneumonectomy for centrally located lung carcinoma. The direct invasion to the descending aorta was only confirmed intraoperatively. After tumour dissection, two aortal cannulas were inserted into the aorta: the first one into the aortic arch over the left subclavian artery, the other one into the descending aorta over the diaphragm. They were then bypassed without a pump. The descending aorta was cross-clamped and replaced by a Dacron graft. There were no early complications : the patient has already survived two years after surgery with no recurrence. PMID- 17405609 TI - Ovarian ligament adenomyoma: a case report. AB - BACKGROUND: Adenomyoma is a benign tumour composed of smooth muscle and benign endometrium. These tumours typically originate within the uterus. An extra uterine adenomyoma is a rare entity. CASE: We report a case of extra-uterine adenomyoma of the ovarian ligament, which was an incidental finding during a total abdominal hysterectomy and bilateral salpingo-oophorectomy for a benign ovarian tumour in a postmenopausal woman. The mass was round with uterine-type smooth muscle and scattered functional endometrial glands and stroma. DISCUSSION: Only seven other cases of an extra-uterine uterine-like mass are reported in the literature. There have been no cases of adenomyoma in the ovarian ligament reported until now. CONCLUSION: It is most likely that this uterine-like mass arose from the tissues of the secondary mullerian system. PMID- 17405610 TI - Symptomatic pseudo-arthrosis of a trapezial ridge fracture. AB - We present a fracture of the ridge of the trapezium with evolution towards a pseudo-arthrosis. PMID- 17405611 TI - History of instrumental haemostasis and the particular contribution of Jules E. Pean. PMID- 17405612 TI - The role of obstetrical ultrasound in screening. PMID- 17405613 TI - Screening for developmental dysplasia of the hip. AB - The reported prevalence of established dislocation the hip in an unscreened population varies from 0.7 to 1.6 / 1000 children in European and American white populations. In clinically screened populations neonatal hip instability is reported to occur in 3 to 30 / 1000 newborns while established congenital dislocation has a prevalence of 0.1- 4/1000 of which 1/1000 is judged to be in need for surgery. Early diagnosis of DDH is essential for successful treatment and later prognosis of the disorder. Combined procedure including evaluation of both hip morphology and hip stability is currently recommended. Opinions differ about the need for universal versus selective sonographic screening for diagnosis of DDH. Currently selective screening of those infants with recognised risk factors and those with abnormal physical examination would be cost-effective and the only practicable method for most countries. PMID- 17405614 TI - Screening for childhood malignancy. AB - Screening for childhood primary malignancy is not a routine practicable issue, due to the rarity of the disease, the absence of specific indicators and the largely unknown causes of childhood cancer. Genetic predisposition may account for as much as 4% of childhood cancer; environmental factors are though to play a smaller role than in the aetiology of adult malignancy: gene-environment interactions could well be important in the induction of malignant disease. Although knowledge about childhood cancer continues to increase, there is much work to be accomplished before reliable preventative measures and screening programme can be recommended. Epidemiologic studies have provided important clues to the etiology of childhood cancer; further insights may be possible by incorporating genomic technology into epidemiologic studies to evaluate cancer susceptibility and to identify high risk children population for cancer. Since second malignancies remain a significant threat to the health of survivors treated for cancer during childhood, vigilant screening is important for those at risk. PMID- 17405615 TI - To screen or not to screen. AB - All screenings lack efficiency because they rely too often on biaised and underpowered statistics. Healthcare business makes tremendous benefits out of well established and easy profit earning processes from screenings to treatments; therefore patients enlightened consent is under influence. In the wealthiest parts of the world, this field of public health sciences impulses studies with intense statistical analysis of tiny differences. About total and cancer specific death rates, meaningful studies should involve over 1,000,000 people. First, who is going to pay for such studies and, if it takes so many people to be sure, the benefit must be very small. In our lifetime, we believed in many concepts only to realise how wrong we were. This will probably be the case for cancer screening. PMID- 17405616 TI - Milestones in paediatric radiology. AB - A historical review of paediatric radiology is presented in the light of the 100th anniversary of the Royal Belgian Rodiological Society and the 25th anniversary of Belgian Paediatric Radiology. Key events in the evolution of paediatric radiology are summarized: the early years of radiology on children, the different measurements used to avoid harmful x-ray effects and up to date medical imaging. Some of the most important associations of radiopaediatrics are mentioned, as well as the importance of immobilization of the young patient resulting in specific x-ray instruments. PMID- 17405617 TI - An unusual presentation of pancreatic serous cystadenoma with acute hemorrhage. AB - Pancreatic serous cystadenomas are benign cystic lesions without complication risks. They can be discovered after investigations for upper abdominal discomfort because of mass effect or incidentally. In rare cases they are detected because of biliary obstruction. Hemorrhagic complication is very unusual. PMID- 17405618 TI - Oro-facio-digital syndrome type 1: imaging findings in an 11-year-old girl. AB - We describe a uniquely illustrated case of orofaciodigital syndrome type 1. We document many of the possible features of this disorder in a young girl using multiple imaging modalities. Furthermore, we prove that the cysts in kidney and liver are not in contact with the excretory system using MRI, something that to our knowledge has not been previously reported. PMID- 17405619 TI - Neurosarcoidosis with bilateral subependymal periventricular lesions. PMID- 17405620 TI - Multiple growing skull fractures. PMID- 17405621 TI - Communication from the European Society of Radiology (ESR) and European Society for Magnetic Resonance in Medicine and Biology (ESMRMB). PMID- 17405622 TI - The in vitro effect of six antimicrobials against Mycoplasma putrefaciens, Mycoplasma mycoides subsp. mycoides LC and Mycoplasma capricolum subsp. capricolum isolated from sheep and goats in Jordan. AB - Respiratory disease in sheep and goats is a major problem in Jordan and is often associated with Mycoplasma species. Without effective vaccines, control is mainly by chemotherapy, but the uncontrolled use of antimicrobials has led to concerns about the potential development of antimicrobial resistance. The in vitro effect of chloramphenicol, florfenicol, enrofloxacin, tylosin, erythromycin and oxytetracycline was determined against 32 isolates of Mycoplasma species-M. mycoides subsp. mycoides LC (6), M. capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions of Jordan. The antimicrobial susceptibility showed some Mycoplasma species-specific differences, with M. capricolum subsp. capricolum being more susceptible to tylosin and erythromycin. Chloramphenicol and florfenicol were the least effective for all three Mycoplasma species. No trends or significant differences in antimicrobial susceptibilities were observed between sheep and goat isolates, between milk or nasal swab isolates, or between isolates from different regions of Jordan. Some isolates of M. capricolum subsp. capricolum and M. putrefaciens showed higher MIC levels with oxytetracycline, as did two isolates of M. mycoides subsp. mycoides LC with tylosin, possibly indicating signs of development of antimicrobial resistance. PMID- 17405623 TI - Perception of cattle farmers of the efficacy of east coast fever immunization in southern Zambia. AB - A study using a structured questionnaire was conducted to assess the perception of cattle farmers of the efficacy of East Coast fever (ECF) immunization in southern Zambia. One hundred and seventy-nine farmers from five districts in southern Zambia were interviewed. The majority of farmers (85%) perceived ECF immunization as being very effective and about half of them (51.4%) preferred immunization to other ECF control strategies. The study showed that the number of calves immunized was strongly associated with the farmer's perception of the benefits of immunization. There was no association between the number of calves immunized and the number of veterinary assistants in a given district or their transport situation. Overall mortality in ECF-immunized calves from various causes stood at 4.2%. Based on farmers' reports, the majority of these deaths seemed to have been caused by anaplasmosis, another tick-borne disease, and might have resulted from relaxation of tick control after ECF immunization. The reasons identified by farmers for not immunizing their animals included failure by immunizing teams to reach certain areas, not having calves of immunization age, and lack of money. These findings provide valuable information on how livestock farmers perceive and adopt new animal disease control strategies and the information could be useful in their planning and implementation. PMID- 17405624 TI - Parasites of the camel in Burkina Faso. PMID- 17405625 TI - Seasonal differences in the incidence of infection with Fasciola gigantica in Cambodian cattle. AB - Farmer's cattle were treated with triclabendazole and used as tracer animals to detect new infections with Fasciola gigantica in three villages located on the bank of the Bassac River (a major tributary of the Mekong River) and in a fourth village located on farmland away from the river, from April 1999 until January 2001. The month of infection was estimated by subtracting 4 months from the date when eggs of F. gigantica were detected in faeces. Farmers were interviewed each month to record the nature of the agricultural and animal husbandry activities that occurred during the previous month, especially events that might have exposed cattle to infection with F. gigantica. Results support the conclusions that infection of cattle in riverbank villages acquired from about August or September until November originated from herbage and water in irrigation canals and dams on the riverbank, and that the progressively increasing monthly incidence from December until April (up to 87% per month in April 2000) was derived from herbage and water in recently harvested rice fields and lakes adjacent to the riverbank. The abrupt cessation of new infection in riverbank villages in May coincided with flooding of low-lying land, the movement of cattle to land above flood height on the riverbank, and a change of diet to dry-land crop residues, stored dry rice stalks, and herbage and water that were unlikely to contain metacercariae. It was concluded that snails in dams and canals on the riverbank became infected with E gigantica after cattle were moved to the riverbank in May, and cercariae shed from these snails provided the new infections that occurred in cattle in August and September. In the village located away from the river, infection of cattle between September and March coincided with the rice harvest, supporting the conclusion that feeding of fresh rice stalks and stubble after the rice was harvested was the main source of infection. The low monthly incidence observed (up to 6.4% per month) was consistent with the hypothesis that snails did not survive in the dry rice fields between crops and that few snails would have been available from the small number of aquatic refuges that persisted through the dry season to recolonize rice fields during the wet season. Between April and August there was no opportunity for new infection because cattle were fed forage from around houses and headlands, and on dry-land crop residues and stored dry rice stalks. Control of fasciolosis was proposed using a single treatment of cattle with triclabendazole in riverbank villages in May when cattle were moved to the riverbank, and after harvest of the last rice field in villages located away from the river. PMID- 17405626 TI - Evaluation of a preliminary title to protect zero-grazed dairy cattle with insecticide-treated mosquito netting in western Kenya. AB - The incidence of trypanosome infection was monitored in dairy cattle during a 6 month trial in Busia and Teso districts, western Kenya, to assess the efficacy of insecticide-treated netting for protection against tsetse flies. Frequently, the fragile netting did not last longer than 2 months because of destruction by strong wind or animal movements. Also, many farmers let their cattle graze freely outside the units during the day, despite technical advice, resulting in exposure of the free-ranging animals to habitats suitable for tsetse and thereby an increased risk of trypanosome infections. The trial groups thus comprised 34 animals from 11 dairy units that were continuously protected, and 153 animals from 46 dairy units that were partially protected. The control group consisted of 162 animals in 42 unprotected units. The phase-contrast buffy-coat technique was used for parasitological monitoring. The mean hazard rate for trypanosomes was significantly lower in protected cows, with a value of 0.007 as opposed to 0.02 for the control animals. Mean packed cell volumes (PCV) were significantly higher in protected cattle (29.7%) than in unprotected ones (27.6%). Farmers with protected animals also reported fewer nuisance flies and mosquitoes in their compounds. PMID- 17405627 TI - Comparative tick counts on game, cattle and sheep on a working game ranch in Kenya. AB - The main objective of the study was to determine the possible influence of host species on the maintenance of ticks in the field by determining the relative contribution of game animals compared to domestic animals. The study was carried out on a game ranch 32 km south-east of Nairobi. Tick counts were carried out on 30 Zebu cattle (Bos indicus) aged 1 to 3 years and 20 red Maasai sheep (Ovis aries) aged 6 months to 1 year grazing with game animals in a common area for a period of 2 years and these counts were compared with those on eland and Thomson's gazelle. Half-body counts were carried out on the cattle and sheep once every week. To avoid excessive stress, the animals were dipped in amitraz whenever the half body counts exceeded 50 fully engorged female ticks of any species. Tick counts on two wild animal species (eland (Taurotragus oryx) and Thomson's gazelle (Gazella thomsonii)) were carried out during the weekly culling of the herbivores. The results revealed that there was no significant difference in the number ticks per square metre between the wild ungulates and the domestic animals. PMID- 17405628 TI - Host differences in response to trickle infection with Fasciola gigantica in buffalo, Ongole and Bali calves. AB - Progressive weight gain, faecal egg counts, packed cell volume, percent eosinophils in blood, serum antibody and serum levels of glutamate dehydrogenase and gamma-glutamyl transpeptidase were recorded in seven swamp buffalo (Bubalis bubalis), 7 Ongole (Bos indicus) and four Bali calves (Bos sundiacus) which were infected orally with 15 metacercariae of Fasciola gigantica twice weekly for 32 weeks. Similar observations were made on four buffalo, 4 Ongole calves and 3 Bali calves maintained fluke-free as controls. Flukes were counted at slaughter 36 weeks after initial infection. Mean daily weight gains of infected Bali (228 +/- 100 (SD) g/day) and infected Ongole calves (328 +/- 57 (SD) g/day) were lower (p = 0.026 and 0.067, respectively) than those of control calves (405 +/- 107 (SD) g/day), but infected buffalo calves (379 +/- 78 (SD) g/day) had similar weight gains to those of the controls (p = 0.57). Throughout the trial, faecal Fasciola egg counts in buffaloes were about one-fifth of counts of Ongole calves, and counts in Bali calves were intermediate. Ongole calves had three times the number of flukes at slaughter in their liver compared to buffalo and Bali calves, which had similar numbers. However, there was evidence that Bali calves had acquired a degree of resistance about 24 weeks after infection commenced and may have lost adult flukes as a consequence. PMID- 17405629 TI - Examination of non-genetic factors affecting the growth performance of djallonke sheep in soudanian zone at the Okpara breeding farm of Benin. AB - A study was carried out at the Okpara breeding farm in Benin. Records on 558 lambs of the Djallonke sheep breed, born from 1997 to 2000, were used to determine the weekly growth curve from birth up to 119 days of age. Non-genetic factors affecting lamb weights at birth and at 30, 60, 90 and 120 days and average daily gains from 0 to 30 days, from 0 to 60 days, from 0 to 90 days and from 0 to 120 days were investigated. A linear model comprising the fixed effects of birth season, birth year, parity number of ewe and sex of lambs was used for analysis of variance by least squares. All fixed effects were significant for weight at 30 and 60 days and average daily gains at 30 days, with the exception of the parity of the ewe for birth weight, weight and average daily gain at 90 days. All fixed effects tended to cancel out for weight and average daily gains by 120 days. Lambs born in the dry season were the heaviest. Single lambs and male lambs were heavier than twins and females. Lambs from ewes of 3rd and 4th parity were heavier, with higher, average daily gains. PMID- 17405630 TI - The small east african shorthorn zebu cows in Kenya. I: linear body measurements. AB - Assessment of genetic diversity is a prerequisite for the management and conservation of animal genetic resources. Appropriate design of breeding programmes is therefore impossible for breeds that have not been adequately characterized either phenotypically and/or genetically. Phenotypic characteristics are important in breed identification and classification in ways that farming communities can relate with. This study phenotypically characterized two breeds of zebu cattle in Kenya. A total of 12 measurements (face length, ear length, horn length, heart girth, height at withers, chest depth, body length, height at rump, pelvis width, corpus length, pin bone width and tail length) were collected on 373 Maasai and 277 Kamba zebu kept by traditional farmers in south east Kenya. The data were classified on the basis of breed group, age group, sex and coat colour pattern. Breed group, age group and sex significantly influenced all measurements. Coat colour pattern significantly influenced only height at withers, corpus length, ear length and tail length. Except for horn and ear length, all the other measurements were significantly higher for the Maasai zebu. Additionally, the Maasai zebu was taller than it was long. The opposite was true for the Kamba zebu. The Maasai and Kamba zebus can be classified as medium-sized breeds; however, great variations exist in their body sizes within and between the breeds. PMID- 17405631 TI - The effect of extended grazing time and supplementary forage on the dry matter intake and foraging behaviour of cattle kept under traditional african grazing systems. AB - An experiment was carried out at Alemaya University in Ethiopia to investigate the effect of night kraaling on the dry matter intake (DMI), live weight gain (LWG) and foraging behaviour of Ogaden cattle. Three groups of four animals were given either 7 h access to pasture per day, simulating traditional grazing (TG) practice; extended grazing (EG) access for 11 h per day; or traditional grazing access plus a nocturnal forage supplement (TF). Live weight gain, DMI and foraging behaviour were measured during the late dry season (EP1) and the wet season (EP2). None of the treatments had any significant effect on either DMI or LWG during EP1 or EP2. Extending pasture access time from 7 h to 11 h did not significantly increase the amount time spent grazing, but grazing intensity was significantly (p < 0.05) reduced during the non-common grazing hours. Step rate was significantly lower (p < 0.01) during EP2 than during EP1 and bites per step were significantly higher (p < 0.001) during EP2 than EP1, indicating that animals had to travel a shorter distance before selecting material to eat during the wet season (EP2). Providing supplementary forage (TF) had no significant effect on any measured parameter. In this study neither of the two low-cost methods (EG and TF) of improving access to forage had any beneficial effect on cattle productivity. It is concluded that, under the prevailing conditions, the traditional grazing practices of this part of Ethiopia do provide sufficient pasture access time to achieve daily voluntary food intake. PMID- 17405632 TI - Radioimmunoassay of milk progesterone as a tool for fertility control in smallholder dairy farms. AB - This study focused on the use of radioimmunoassay of progesterone in milk for the diagnosis of post-partum ovarian cyclicity and accurate detection of oestrus and non-pregnancy in cows in the artificial insemination (AI) programme in Bangladesh. In Investigation 1, milk samples were collected on day 0 (day of AI), day 9-13 and day 21-24 from 444 milking cows of various breeds presented for the first postpartum insemination by 413 farmers living at 182 villages/regions in Mymensingh District from 6 AI centres and sub-centres. Each cow was then examined three times after each AI until it stopped returning to oestrus. Sixty to 90 days after the last AI, the cows were examined per rectum to confirm the pregnancy. Milk progesterone data on day 21-24 contributed to a clear diagnosis with respect to non-pregnancy in 100% cows, indicating a possible use of this progesterone assay for identifying non-pregnant cows in AI programmes. In Investigation 2, milk progesterone was monitored two times in a month with a 10-day interval in 88 cows. The samples were taken between 10 days after calving and the first detected oestrus, followed by two more samples 10 days apart. The proportion of cows accurately detected in oestrus was 30%. Another 30% were stated to be in oestrus when they were not (false positive) and 40% were not detected when they were in oestrus (false negative). The mean intervals between calving and oestrus and between calving luteal activity were 40 to 362 days (median=120, n=82) and 34 to 398 (median=111, n=64) days, respectively. The body condition scores at calving and at the initiation of luteal activity influenced the interval between calving and luteal activity (p < 0.05). Cows suckled twice daily initiated luteal activity earlier than their counterparts suckled several times daily (p < 0.05). Determination of progesterone in milk on day 21-24 is a good means for detecting non-pregnant cows. PMID- 17405633 TI - Surgical management of radial head fractures. PMID- 17405634 TI - Maintenance of reduction of pediatric distal radius fractures with a sugar-tong splint. AB - Distal radius fractures are common injuries in children. Displaced fractures have traditionally been treated with closed reduction followed by immobilization in a long arm cast. Because of variable success rates with this technique, a trend in the literature is toward operative fixation of these fractures. A popular alternative practice involves temporary immobilization in a sugar-tong splint, though we are unaware of any studies demonstrating the efficacy of this technique in children. We present our experience in treating these injuries initially with a sugar-tong splint and then with a short arm cast. We retrospectively reviewed the cases of 53 patients (age range, 2-12 years) treated with closed reduction and a sugar-tong splint followed by conversion to a short arm cast after 2 to 3 weeks. In 51 (96%) of 53 fractures, reduction was maintained without more aggressive intervention. The sugar-tong splint is effective in maintaining reductions in pediatric distal radius fractures and has none of the added risks associated with current alternative methods. PMID- 17405635 TI - Health care technology assessment: implications for modern medical practice. Part II. Decision making on technology adoption. AB - Health care technology assessment, the multidisciplinary evaluation of clinical and economic aspects of technology, has come to have an increasingly important role in health policy and clinical decision-making. In Part I--Understanding Technology Adoption and Analyses--this review addressed the difficult challenges posed by assessment and provided a guide to the methodologies used. Part II presents the factors that drive the technology choices made by patients, by individual physicians, by provider groups, and by hospital administrators. PMID- 17405636 TI - Clinical decision making: doctor, when can I drive? AB - The orthopedic surgeon has little evidence-based literature to refer to when deciding when a patient is "safe" to drive a car. Current advice is derived from several empirical reports on soft-tissue and bone healing. Using the terms surgery and driving, I searched Pubmed and Medline to review the relevant English language research literature published between 1960 and the present. Of the 975 studies found and examined--all studies, surveys, and case reports involving the extremities were considered--35 had orthopedic relevance. Recommendations for return to driving are presented here. Orthopedic surgeons should find this review an aid when they use the literature to make informed decisions about patients' return to driving and when they conduct further evidence-based investigation to determine orthopedic fitness to drive. PMID- 17405637 TI - Closed reduction for treatment of developmental dysplasia of the hip in children. AB - Although many studies have analyzed the success rates of closed reduction and spica casting for the treatment of developmental dysplasia of the hip (DDH) in children, the definition of success for this procedure is not standardized in the literature. We retrospectively reviewed our experience with closed reduction for treatment of DDH in 30 children (35 hips) over an 8-year period to determine the success rates of this procedure on the basis of how success is defined. In only 1 patient (2 hips, 6%) were the hips unable to be concentrically reduced with sufficient stability at the time of closed reduction. In 10 (30%) of the other 33 hips, the acetabulum failed to develop sufficiently after closed reduction, and a secondary surgery was required a mean of 22 months after cast removal. Four (12%) of the 33 hips developed radiographic evidence of avascular necrosis. Therefore, the success rate of closed reduction for DDH varies markedly depending on how success is defined. PMID- 17405638 TI - Type III acromioclavicular separation: results of a recent survey on its management. AB - The issue of managing type III acromioclavicular (AC) separations remains controversial, and decisions about using operative versus conservative management have undergone many distinct changes over the years. To review current management preferences within the orthopedic community, we sent a mail-in survey to all members of the American Orthopaedic Society for Sports Medicine (AOSSM) and approved Accreditation Council for Graduate Medical Education (ACGME) orthopedic program residency directors. Of the 664 respondents (577 AOSSM members, 87 directors), 81% (71/87 AOSSM members) to 86% (502/577 directors) continue to treat uncomplicated type III AC separations conservatively. Providing a sling for comfort remains the preferred type of conservative management (AOSSM members, 91% [456/502]; directors, 89% [63/71]). For surgical management, respondents recommended resection of the distal clavicle slightly more often than not (AOSSM members, 57% [42/74]; directors, 59% [319/538]) and rigid stabilization of the AC joint during early postoperative rehabilitation (AOSSM members, 80% [444/555]; directors, 82% [61/74]). Finally, most recommended reconstructing either the coracoclavicular ligaments (69% [330/476] and 61% [33/54], respectively) or both the coracoclavicular ligaments and the AC ligaments (27% 130/476] and 33% [18/54]) when addressing this problem. Since the early 1990s, there has been little change in initial conservative management of type III AC separations. Furthermore, the surgical approach to reconstruction, when necessary, has also undergone relatively few changes, with the exception of an increased preference for primary distal clavicle excision. PMID- 17405639 TI - Making a difference, one member at a time. PMID- 17405640 TI - RHIOs and the value proposition. Value is in the eye of the beholder. PMID- 17405641 TI - This year's models. A look at patient ID in the four newly demonstrated NHIN prototypes. PMID- 17405642 TI - Nebraska's far-flung HIEs move forward. PMID- 17405643 TI - HIE at the state level. A role for states in regional data-sharing networks? PMID- 17405644 TI - Plugging in physician practices. Software implementation offers insights on EHRS, community networking. PMID- 17405645 TI - The legal health record. A component of overall EHR strategy. PMID- 17405646 TI - Lessons from Hurricane Katrina. One HIM director's experience. PMID- 17405647 TI - Linking anti-fraud and legal EHR functions. PMID- 17405648 TI - Disclosure of medical record copies for payment purposes. PMID- 17405649 TI - Assessing and improving EHR data quality. PMID- 17405650 TI - Medicare reimbursement changes for ASCs. PMID- 17405651 TI - Coding guidelines for pain. PMID- 17405652 TI - Putting the HIM stamp on RHIOs. PMID- 17405653 TI - [Effects of two types of treatment for shoulder subluxation in hemiplegics: a pilot study]. AB - PURPOSE: The aim of this pilot study is to compare the effects of a new sling, a special type of strapping and their association to treat shoulder subluxation. MATERIALS AND METHODS: The effectiveness of a modified version of a sling, a special type of strapping and their association to treat shoulder subluxation has been compared in three hemiplegic patients. Shoulder subluxation has been diagnosed by palpation, then assessed by radiographic measurements in sitting position with the plane of the scapula method (at 30 degrees to the coronal plane). The distance between the inferior acromial surface and the horizontal line crossing through the central point of the humeral head (in millimeters) has been taken as measurement of subluxation. RESULTS: All the patients showed a reduction of the acromial-humeral distance when the sling was applied with strapping. When sling or strapping alone was applied, a slight difference was measured in each patient. CONCLUSIONS: The association of sling and strapping seems to be an effective strategy to treat shoulder subluxation in hemiplegic patients in sub-acute stage. The tools described in the paper give a better cost/benefit ratio in comparison with others described in literature and widely used in clinical practice. Hawever, further investigations are necessary to confirm these results. PMID- 17405654 TI - [Possible effects of non invasive mechanical ventilation on respiratory drive and muscles]. AB - OBJECTIVE: To evaluate whether long-term Non-Invasive Mechanical Ventilation (NIMV) might have an effect on respiratory drive and respiratory muscles strength, measuring mouth occlusion pressure (P0,) and maximal inspiratory pressure (MIP). PATIENTS AND METHODS: 20 consecutive patients with hypercapnic respiratory failure underwent measurements of dyspnea, respiratory drive and respiratory muscles strength before hospital treatment with NIMV; those patients who showed significant improvement of gas-exchange continued home ventilation for a period of four weeks, and were readmitted to hospital for re-evaluation of Borg's scale for dyspnea, P0,1 and MIP. RESULTS: Data obtained show a mild reduction of P0,1 and a significant improvement of respiratory muscles strength, with satisfactory dyspnea relief. CONCLUSIONS: We conclude that unloading respiratory muscles through mechanical ventilation results in better muscle performance in the long-term that could act, together with normalization of gas exchange, on neuromuscular respiratory drive and contribute to dyspnea relief. PMID- 17405655 TI - [Osteoporosis and anorexia nervosa: ultrasonometric and biochemical evaluation- preliminary data]. AB - AIMS: To evaluate the relationship between Anorexia Nervosa (AN) and Osteoporosis (OP) by bone heel ultrasonometric measurement and biochemical bone metabolic data. MATERIALS AND METHODS: AN, 15 patients (2 males / 13 females; mean age 27.5 yr, range 16-44; mean BMI 15.78, range 13-19.3); normal subjects (NS), 10 (2 males / 8 females; mean age 27.5 yr, range 17-40; mean BMI 26.2, range 25-28). An Hologic Sahara ultrasound heel instrument has been utilized in order to obtain the following parameters: broadband ultrasound attenuation (BUA), speed of sound (SOS), Stiffness' index, bone mineral density (BMD) and T-score. As markers of bone formation osteocalcin (OC, ng/ml) and of resorption pyridinolines (Pyr, pmol/umol creatinine e deoxy-Pyr) have been studied by standardized analytical methods. RESULTS: (expressed as x+/-SD). PATIENTS: BUA, 65+/-11.22 (p<0.01); SOS, 1544.14+/-73.5 (ns); Stiffness, 89.8+/-19.4 (p<0.01): BMD, 0.55+/-0.53 (p<0.01); T-score, -1.4+/-1.12 (p<0.01); OC, 4.05+/-2.3 (p<0.01); Pyr, 53+/-21 (ns); d-Pyr, 7.17+/-4.5 (ns). NS: BUA, 88.57+/-8.63; SOS, 1567.72+/-11.88; Stiffness, 108.07+/ 4.97; BMD, 0.611+/-0.027; T-score, 0.22+/-0.3; OC, 8.5+/-4.5; Pyr, 60+/-25; d Pyr, 8.5+/-3.5. CONCLUSIONS: Our data seem to confirm that AN represents an important risk factor for OP. The ultrasonometric data in AN patients document some statistically significant differences from SN in term of BMD and T-score reduction. The metabolic data show that OC is reduced in AN patients, on the contrary, no differences appear in term of resorption bone markers. PMID- 17405656 TI - [On therapeutic use of stem cells and the sources of morality]. AB - The Article deals with the ethical issues around the therapeutic utilization of stem cells (SC), particularly facing with factors of morality: i.e., object of the human action, aims and circumstances. Crucial ethical questions are recognized in the last factor (circumstances), particularly: the origin and the availability of SC, the consequences of the therapeutic action through SC, the diffusion/communication of scientific data around the clinical use of SC. The paper assumes the perspective of the ethics considered as practical-normative science, centered on the human person (as a whole) as the value of reference. PMID- 17405657 TI - [Ulcerative colitis and thrombocytopenia: a clinical case of fortuitous association?]. AB - Ulcerative colitis (UC) is an inflammatory bowel disease of unknown etiology, involving primarily the rectum with major intestinal symptoms. Additionally, UC is often associated with extraintestinal manifestations, especially arthropathies, as well as with some autoimmune disorders. Vice versa, UC is rarely described in association with hematologic abnormalities, such as autoimmune hemolytic anemia and immune thrombocytopenic purpura with antiplatelet antibodies positive. Usually UC precedes the onset of thrombocytopenia by days or years or coincides with it. We report a case of UC and thrombocytopenia with negative anti-platelet antibodies in which an immunosuppressive therapy with corticosteroids obtained significant remission of intestinal symptoms along with a rapid increase of platelet count. PMID- 17405658 TI - Relationships between 24h observations in intraocular pressure vs blood pressure, heart rate, nitric oxide and age in the medical chronobiology aging project. AB - OBJECTIVE: To evaluate associations between intraocular pressure (IOP) and blood pressure (BP), heart rate (HR), serum nitric oxide (NO), diurnal variations, diabetes and aging in data collected during 24h studies of men conducted over 34y. MATERIALS AND METHODS: As part of the Medical Chronobiology Aging Project, male Army veterans, ages 22 to 81y, without a history of eye disease, were studied around-the-clock in May 1969 (n = 13), 1979 (n = 11), 1988 (n = 11), 1993 (n = 11), 1998 (n =12) and 2003 (n = 10). Measurements of IOP (R & L eyes, supine position), BP and HR (sitting position), and collection of blood were obtained every 3h (8 readings/24h) from 19:00h to 16:00h the next day. Individual time series were analyzed for circadian characteristics by the least-squares fit of a 24& 12h cosine. After normalizing all data to percent of mean to reduce inter subject variability in levels, grouped data were analyzed for time-effect by ANOVA and for circadian rhythm by multiple component (24h&12h) cosine fitting. Individual 24h averages were analyzed by simple and multiple regression for relationships between IOP and systemic variables, diabetic status and age. RESULTS: Over the 34y study span, 22 men provided sixty-three 24h profiles for IOP & HR, 61 for BP, and 21 for NO. Using all normalized data, a significant circadian rhythm was found for each variable at p <0.001. Circadian peaks (orthophases) are located in the late morning for IOP-R (10:20h) and IOP-L (10:52h), and in the evening for HR (18:52h), NO (20:00h), SBP (20:40h) and DBP (21:44h). An out-of-phase relationship of about 10h is noted on a group basis between IOP vs BP, HR and NO. The locations of individual circadian peaks for IOP R were found around the clock, but with a significant predominance between 10:00 and 16:00h (day type), and 04:00-10:00h (morning type). In contrast, BP, HR and NO showed a significant clustering of evening type or night type peaks. The overall mean IOP for the right eye was slightly, but not significantly, higher than the left eye (17.60+/-0.21 vs 17.34+/-0.18 mmHg; p = 0.385), with a strong positive correlation between both eyes (R = 0.952, p <0.0001). IOP showed a significant positive correlation with SBP (R = 0.49, p <0.001), diabetic status (R = 0.47, p <0.001), age (R = 0.32, p = 0.011), and HR (R = 0.28, p = 0.031). A multiple regression using SBP, DBP, HR, age and diabetic status (5 men became diabetic over the 34y study span) as independent variables resulted in SBP being the strongest predictor of IOP (p = 0.0001), followed by DBP (p = 0.0103). After adjustment for BP, independent effects of age (p = 0.187), HR (p = 0.789) and diabetic status (p = 0.153) were eliminated from the prediction equation. CONCLUSIONS: The results of these studies reveal significant circadian variations in IOP, BP, HR and NO, with peak levels, on average, near noon for IOP and in the evening for BP, HR and NO. An increase in SBP was associated with an increase in IOP. While SBP and DBP are significant predictors of IOP levels, single measurements during regular clinic hours may not reveal the full functional relationship between the variables measured in our studies. Therefore, circadian information on total 24h patterns may contribute to the reliability of diagnosis and guide proper individualized timing of optimal patient management (e.g., for glaucoma, hypertension, diabetes, among other conditions). PMID- 17405659 TI - [Diet and nutrition in nephrolitiasis]. AB - Nephrolitiasis is a frequent metabolic disease, with a high rate of recurrences. Epidemiological studies reveal that about 80% of all kidney stones are composed of calcium salts (75% calcium oxalate), while about 5% are pure uric acid. Urolithiasis is a multifactorial disease with several underlying disorders of metabolism: that is why diet is an important treatment, especially in the prevention of recurrences. Nutritional intervention is based on a high water intake, physiological calcium intake, modest sodium and animal protein restriction and vitamin C intake <2 gr daily. In case of diagnosed disorders of specific metabolic pathways, a low oxalate, low purine-diet should be advisable. PMID- 17405660 TI - [New combination chemotherapy regimens in the primary treatment of operable breast cancer]. AB - Primary (neoadjuvant) systemic chemotherapy is the standard treatment for locally advanced breast cancer and a standard option for primary operable disease. Although survival results are similar, primary chemotherapy has the following advantages in comparison to adjuvant chemotherapy: it represents a chemosensitivity test in vivo and can be of value in determining the prognosis of the patient since pathologic complete responses are related to improved survival. Among a variety of primary chemotherapy regimens currently available, the most effective seem to be those containing both anthracyclines and taxanes, expecially when these agents are administered sequentially. There are also several ongoing studies evaluating primary hormonal therapy and the combination of cytotoxic chemotherapy and targeted agents. It is conceivable that in the future primary chemotherapy of breast cancer will be increasingly used. In fact, besides its clinical effectiveness, primary chemotherapy is extremely important to evaluate new agents and to find useful prognostic and predictive factors. PMID- 17405661 TI - [Gastroesophageal reflux disease: clinical and pathophysiological features (part I)]. AB - Gastroesophageal reflux disease (GERD) can be described as a clinical picture resulting from the reflux of stomach contents into the esophagus. The actual prevalence of GERD remains unestablished, although this disorder is generally common in old patients, male sex and in subsets of patients with pulmonary manifestations such as asthma. From a pathophysiological stand-point, GERD is thought to have a multifactorial etiology which involves genetics, anatomical, functional, environmental, hormonal and pharmacological factors. GERD has different clinical presentations which may be divided in three main classes: typical symptoms (heartburn and regurgitation); atypical or extraesophageal symptoms (angina-like chest pain, asthma, chronic cough and laryngitis); and complications (ulcers, strictures and Barrett's esophagus). In GERD diagnosis a key role is played by: accurate symptom evaluation, response to proton pump inhibitors and, finally, at least one in a life-time endoscopy. Moreover, barium swallow X-ray, 24-h esophageal pH monitoring and gastro-esophageal manometry can be useful to support diagnosis in some unusual cases or in cases partially or unresponsive to standard pharmacologic treatment. PMID- 17405662 TI - [First episode psychoses: psychosocial, interpersonal and cognitive bases for a therapeutic approach]. AB - The occurrence of psychotic symptoms in first episode psychoses is the ultimate stage of a series of behavioural, cognitive and psychosocial signs. Among these, only a small set are pathognomonics, but more importantly, they continue to operate and to modify the clinical picture. The instability of diagnosis and outcome require a large series of multimodal and atheoretical treatment approaches. Interventions should be flexible, acceptable, intrinsically effective and evidence based. Clinical and therapeutic decisional algorithms will be presented to be adopted in mental health services for people at first episode psychoses. PMID- 17405663 TI - Modern neuropsychology in France: Francois Lhermitte (1921-1998). PMID- 17405664 TI - A double dissociation between impact factor and cited half life. PMID- 17405665 TI - Autobiographical memory, the sense of recollection and executive functions after severe traumatic brain injury. AB - Residual disorders of autobiographical memory long after trauma resulting from head injury are rarely assessed, even though they may affect social adjustment and the resumption of daily life. We conducted a thorough study of autobiographical memory in severe traumatic brain injury (TBI) patients, examined at least one year post-trauma. Twenty-five patients were submitted to a novel and controlled autobiographical procedure specially designed to measure episodic memories (i.e., unique, specific in time and space, and detailed) from their entire life span with two kinds of self-remembering experience. The ability to mentally travel back through time and re-experience the source of acquisition, i.e. autonoetic consciousness, was assessed via the "Remember/Know" paradigm and a checking procedure of sense of remembering. Self-perspective in visual imagery, which is also critically involved in episodic recollection, was assessed by the "Field/Observer perspective" paradigm. In addition, the patients underwent a battery of standardized neuropsychological tests to assess episodic and semantic memory, orientation and executive functions. The results showed that the patients, compared with healthy controls, were significantly impaired in recalling episodic autobiographical memories. This impairment was not related to the life period tested or the patients' ages nor the intellectual impairment. Deficits involved disturbances in sense of remembering, visual imagery self perspective and recollection of spatiotemporal details. Stepwise-regression analyses carried out in the TBI patients revealed a significant relationship between an abnormal sense of remembering and executive dysfunction covering both anterograde and retrograde components. The novel assessment used in this study provides the first detailed evidence of a more fine-grained deficit of autobiographical memory in TBI patients. Indeed, the results suggest that these patients, long after trauma, present autonoetic consciousness and self perspective disorders, which include sense of identity (the self) as a continuous entity across time, probably related to frontal dysfunction. PMID- 17405666 TI - Processing of facial blends of emotion: support for right hemisphere cognitive aging. AB - Clinical research on facial emotions has focused primarily on differences between right and left hemiface. Social psychology, however, has suggested that differences between upper versus lower facial displays may be more important, especially during social interactions. We demonstrated previously that upper facial displays are perceived preferentially by the right hemisphere, while lower facial displays are perceived preferentially by the left hemisphere. A marginal age-related effect was observed. The current research expands our original cohort to include 26 elderly individuals over age 62. Fifty-six, strongly right-handed, healthy, adult volunteers were tested tachistoscopically by flashing randomized facial displays of emotion to the right and left visual fields. The stimuli consisted of line drawings displaying various combinations of emotions on the upper and lower face. The subjects were tested under two conditions: without attend instruction and with instructions to attend to the upper face. Based on linear regression and discriminant analyses modeling age, subject performance could be divided into two distinct groups: Young (< 62 years) and Old (> 62 years). Without attend instructions, both groups robustly identified the emotion displayed on the lower face, regardless of visual field presentation. With instructions to attend to the upper face, the Old group demonstrated a markedly decreased ability to identify upper facial displays, compared to the Young group. The most significant difference was noted in the left visual field/right hemisphere. Our results demonstrate a significant decline in the processing of upper facial emotions by the right hemisphere in older individuals, thus providing partial support for the right hemisphere hypothesis of cognitive aging. The decreased ability to perceive upper facial displays coupled with age-related deficits in processing affective prosody may well cause impaired psychosocial competency in the elderly. PMID- 17405667 TI - Bilateral processing of redundant information: the influence of stimulus notation and processing speed in number comparison. AB - Improved performance is generally observed when identical copies of a stimulus are presented bilaterally to both visual half fields (VHFs), relative to unilateral single presentations. We investigated the influence of stimulus notation and processing speed on this bilateral redundancy gain (B-RG) in number comparison. Two experiments were performed with consistent (i.e., two identical copies of Arabic digits or word numerals) and inconsistent (i.e., an Arabic digit in one VHF with a word numeral of the same value in the other VHF) displays (see also Marks and Hellige, 2003 for a similar design in number naming). In Experiment 1, the processing speeds of the Arabic digits were faster than the ones of the word numerals while in Experiment 2, the processing speeds of the two stimulus notations were kept equal. A significant difference between consistent and inconsistent conditions was restricted to Experiment 1 in which the processing speeds of the two stimulus notations diverged. These results are discussed in the framework of a simple race model in which two independent hemispheric processes compete for the control of response, and the faster process wins. PMID- 17405668 TI - Lower limb sensorimotor network: issues of somatotopy and overlap. AB - Functional magnetic resonance imaging (fMRI) was used (1) to describe the pattern of whole brain activity during motion of isolated joints of the lower limb, (2) to examine the somatotopic organization of lower limb joint representations in the primary sensorimotor cortex and the anterior lobe of the cerebellum and 3) to quantify the degree of overlap between these lower limb joint activations. Eighteen healthy, right leg dominant volunteers participated in a motor block design study, performing repetitive knee, ankle and toes flexion/extension movements. In order to relate lower limb joints activation to the well-described patterns of finger movement, serial finger-to-thumb opposition was also assessed. All movements were auditory paced at 72 beats/min (1.2 Hz). Isolated lower limb joints movement activated a distributed sensorimotor network, including primary and non-primary sensorimotor areas. Although a large overlap was evident in primary sensorimotor cortex (SM1) and cerebellum representations of the three lower limb joints, a somatotopic arrangement was recognizable with reference to center of mass coordinates of each individual joint in the above areas. Detection of active brain regions during movement of the lower limb joints is feasible with fMRI although a carefully optimized methodology protocol is required. PMID- 17405669 TI - Long-term memory for verbal and visual information in Down syndrome and Williams syndrome: performance on the Doors and People test. AB - Previous studies have suggested that Williams syndrome and Down syndrome may be associated with specific short-term memory deficits. Individuals with Williams syndrome perform relatively poorly on tests of visuo-spatial short-term memory and individuals with Down syndrome show a relative deficit on verbal short-term memory tasks. However, these patterns of impairments may reflect the impact of generally impaired visuo-spatial processing skills in Williams syndrome, and verbal abilities in Down syndrome. The current study explored this possibility by assessing long-term memory among 15 individuals with Williams syndrome and 20 individuals with Down syndrome using the Doors and People test, a battery which assesses recall and recognition of verbal and visual information. Individuals' performance was standardised for age and level of intellectual ability with reference to that shown by a sample of 110 typically developing children. The results showed that individuals with Down syndrome have no differential deficits in long-term memory for verbal information, implying that verbal short-term memory deficits in this population are relatively selective. Instead both individuals with Down syndrome and with Williams syndrome showed some evidence of relatively poor performance on tests of long-term memory for visual information. It is therefore possible that visuo-spatial short-term memory deficits that have previously been demonstrated in Williams syndrome may be secondary to more general problems in visuo-spatial processing in this population. PMID- 17405670 TI - Directional disorientation following left retrosplenial hemorrhage: a case report with fMRI studies. AB - We report a 55-year-old right-handed man who presented with topographical disorientation following left retrosplenial hemorrhage. His directional information about familiar places, encoded by previous navigation, was severely impaired, and he could not learn the direction to new places in large-scale spaces beyond the range of visual surveillance. By contrast, he had no difficulties with directional information encoded in a tabletop manner: he could locate major cities or countries on a map, and he also could memorize the spatial relationship of objects in a room. Six months after the ictus, when he had recovered from his directional disorientation, a functional magnetic resonance imaging (fMRI) study of mental navigation demonstrated prominent activation in the retrosplenial area along the right parieto-occipital sulcus and the circumference of the injured area on the left side. The present study, together with previous investigations including clinical case reports, functional neuroimaging, and anatomical and physiological studies on monkeys, suggests that the 'sense of direction' in a large-scale locomotor environment is subserved by the visual area along the parieto-occipital sulcus, and that bilateral deterioration of this function causes directional disorientation. PMID- 17405671 TI - Is short-term memory for discrete arm movements impaired in Huntington's disease? AB - Explicit memory for arm movements was investigated in patients with Huntington's disease (HD) and normal controls (NC). Participants recalled the end-position of arm movements determined by the examiner following an immediate or a 30-second delay that was either unfilled or filled with a motor distractor task. Movement error was measured in absolute (AE), variable (VE), and constant (CE) error. When performance was measured in AE, HD patients were impaired equally across delays compared to controls, regardless of whether the delay was filled or unfilled. HD patients did not demonstrate a directional bias (i.e., CE), but they were more inconsistent in their reproduction of arm movements (i.e., VE). Both groups were more variable in their reproduction of the movements following the delay, and were most variable when the delay was filled with motor distraction. Results preliminarily suggest that the caudate is specifically involved in the encoding, or temporary storage, of explicit motor information and may be less involved in the short-term retention of newly learned motor information. Results should be viewed somewhat cautiously, however, as the relative contributions of proprioceptive sense and primary motor functioning on task performance were not fully explored. PMID- 17405672 TI - "Apperceptive" alexia in posterior cortical atrophy. AB - The most common presenting complaint in posterior cortical atrophy (PCA) is reading difficulty. Although often described as an alexia without agraphia, alexia in PCA may have multiple causes, including a primary visuoperceptual etiology, attentional alexia, and central reading difficulty. This study evaluated 14 patients with early PCA and disturbances in reading ability in comparison to 14 normal controls. All 14 patients had a progressive disorder of complex visual functions and neuroimaging evidence of occipitoparietal dysfunction. They underwent a task requiring identification of single letters with and without flanking distractors. They also read single words consisting of regular English spelling or irregular grapheme-phoneme correspondence (irregular words) and pronounceable nonsense words (pseudowords). The PCA patients made errors in letter identification when letters were flanked by visually similar letters or numbers. They could read most single regular and irregular words but made visual errors and had particular trouble with pseudowords. They could not use a letter-by-letter reading strategy effectively. The PCA patients had similar difficulties on other visuoperceptual tests. These findings are consistent with an alexia manifested by perceptual and attentional difficulty on attempting serial visual processing of letters in the context of other letters. This "apperceptive alexia" results when the configuration of letters into words is impaired during letter-by-letter reading. Disproportionate difficulty reading pseudowords suggests an additional impairment in phonological processing. PCA patients have variable neuropathology and individual patients may have other contributions to their reading impairment. PMID- 17405673 TI - The origin of diseases: musings on ecology and the evolution of medical ideas. PMID- 17405674 TI - Reclaiming the invisible. PMID- 17405675 TI - Factors associated with herbal therapy use by adults in the United States. AB - OBJECTIVE: To examine the patterns of herbal therapy use among adults in the United States and to describe factors associated with herb use. DESIGN: We examined the use of natural herbs from the 2002 National Health Interview Survey (NHIS). We analyzed factors associated with herb use and reasons for herb use with logistic regression. RESULTS: Factors associated with herb use include the following: age (45-64 years old), being uninsured, being female, having a higher education, living in the West, using prescription medications or over-the-counter (OTC) medications, and self-identified as "non-Hispanic other." Factors associated with no herb use include being non-Hispanic black and living in the South or Midwest. Seventy-two percent of those who used herbs used prescription medications, and 84% of those who used herbs also used an OTC medication in the prior 12 months. Among adults who used herbs, the most commonly mentioned were echinacea (41%), ginseng (25%), gingko (22%), and garlic (20%). The most frequent conditions for herb use were head or chest cold (30%), musculoskeletal conditions (16%), and stomach or intestinal illness (11%). Among those who used herbs in the prior year, factors associated with using herbs because conventional medical treatments were too expensive included being uninsured, having poor health, and being 25-44 years old. CONCLUSIONS: Nearly 1 in 5 people in the US population report using an herb for treatment of health conditions and/or health promotion. More than half did not disclose this information to a conventional medical professional. PMID- 17405676 TI - Adverse interactions between herbal and dietary substances and prescription medications: a clinical survey. AB - CONTEXT: Patients often combine prescription medications with herbal and dietary substances (herein referred to as herbal medicines). A variety of potential adverse herb-drug interactions exist based on the pharmacological properties of herbal and prescription medications. OBJECTIVE: To determine the incidence of potential and observed adverse herb-drug interactions in patients using herbal medicines with prescription medications. DESIGN: Consecutive patients were questioned about their use of herbal medicines in 6 outpatient clinics. Patients reporting use of these products provided a list of their prescription medications, which were reviewed for any potential adverse herb-drug interactions using a comprehensive natural medicine database. Any potential adverse herb-drug interactions prompted a review of the patient's chart for evidence of an observed adverse herb-drug interaction. MAIN OUTCOME MEASURE: The rate of potential and observed adverse herb-drug interactions. RESULTS: Eight hundred four patients were surveyed, and 122 (15%) used herbal medicines. Eighty-five potential adverse herb-drug interactions were found in 49 patients (40% of herbal medicine users). Twelve possible adverse herb-drug interactions in 8 patients (7% of herbal medicine users) were observed. In all 12 cases, the severity scores were rated as mild, including 8 cases of hypoglycemia in diabetics taking nopal (prickly pear cactus). CONCLUSIONS: A substantial number of potential adverse herb-drug interactions were detected and a small number of adverse herb-drug interactions observed, particularly in diabetics taking nopal. Screening for herbal medicine usage in 804 patients did not uncover any serious adverse interactions with prescription medications. PMID- 17405677 TI - Evaluating anecdotes and case reports. PMID- 17405678 TI - Antioxidants and other nutrients do not interfere with chemotherapy or radiation therapy and can increase kill and increase survival, Part 2. AB - PURPOSE: Some in the oncology community contend that patients undergoing chemotherapy and/or radiation therapy should not use food supplement antioxidants and other nutrients. Oncologists at an influential oncology institution contended that antioxidants interfere with radiation and some chemotherapies because those modalities kill by generating free radicals that are neutralized by antioxidants, and that folic acid interferes with methotrexate. This is despite the common use of amifostine and dexrazoxane, 2 prescription antioxidants, during chemotherapy and/or radiation therapy. DESIGN: To assess all evidence concerning antioxidant and other nutrients used concomitantly with chemotherapy and/or radiation therapy. The MEDLINE and CANCERLIT databases were searched from 1965 to November 2003 using the words vitamins, antioxidants, chemotherapy, and radiation therapy. Bibliographies of articles were searched. All studies reporting concomitant nutrient use with chemotherapy and/or radiation therapy (280 peer-reviewed articles including 62 in vitro and 218 in vivo) were indiscriminately included. RESULTS: Fifty human clinical randomized or observational trials have been conducted, involving 8,521 patients using beta-carotene; vitamins A, C, and E; selenium; cysteine; B vitamins; vitamin D3; vitamin K3; and glutathione as single agents or in combination. CONCLUSIONS: Since the 1970s, 280 peer-reviewed in vitro and in vivo studies, including 50 human studies involving 8,521 patients, 5,081 of whom were given nutrients, have consistently shown that do not interfere with therapeutic modalities for cancer. Furthermore, non-prescription antioxidants and other nutrients enhance the killing of therapeutic modalities for cancer, decrease their side effects, and protect normal tissue. In 15 human studies, 3,738 patients who took non-prescription antioxidants and other nutrients actually had increased survival. PMID- 17405679 TI - The compliance paradox: what we need to know about "real-world" dietary supplement use in the United States. AB - Current research on dietary supplements (primarily survey-based prevalence studies and clinical trials of safety and efficacy) is inadequate for understanding how consumers use supplements in the real world. Analyzing interview data from formative research with dietary supplement users (N=60), we observed skepticism in the way our informants interpret scientific information about supplements, trust in referrals from those they feel are like them, and experimentation with products in order to tailor them to their bodies and needs. We stress the need for qualitative research focusing on patterns of supplement use in context (rather than as isolated supplements in fixed doses), the network effect of supplement use, and the way information about supplements is translated and transmitted. Furthermore, we urge clinicians to pay careful attention not only to whether patients are taking dietary supplements, but also how supplements are being used alone and in combination with other supplements, pharmaceuticals, and over-the-counter (OTC) medications. PMID- 17405680 TI - Efficacy of a medical nutriment in the treatment of cancer. PMID- 17405681 TI - Jim Duke, PhD: ethnobotanist extraordinaire. Interview by Frank Lampe and Suzanne Snyder. PMID- 17405682 TI - Managing biotransformation: introduction and overview. PMID- 17405683 TI - Who ignores individuality fails the patient. PMID- 17405684 TI - Detoxification basics. PMID- 17405685 TI - Diet and detoxification enzymes. PMID- 17405686 TI - Working up the toxic patient: practical intervention and treatment stategies. PMID- 17405687 TI - Diet, genetic polymorphisms, detoxification, and health risks. AB - Modulation of detoxification enzymes is one mechanism by which diet may influence risk of cancer and other diseases. However, genetic differences in taste preference, food tolerance, nutrient absorption, and metabolism and response of target tissues all potentially influence the effect of diet on disease risk. Thus, disease prevention at the individual and population level needs to be evaluated in the context of the totality of genetic background and exposures to both causative agents and chemopreventive compounds. Polymorphisms in the detoxification enzymes that alter protein expression and/or function can modify risk in individuals exposed to the relevant substrates. Diet is a mixture of carcinogens, mutagens, and protective agents that are all metabolized by detoxification enzymes. Genotypes associated with more favorable handling of carcinogens may be associated with less favorable handling of phytochemicals. For example, glutathione S-transferases (GST) detoxify polycyclic aromatic hydrocarbons present in grilled meats. GSTs also conjugate isothiocyanates, the chemopreventive compounds found in cruciferous vegetables. Polymorphisms in the GSTM1 and GSTT1 genes result in complete lack of GSTM1-1 and GSTT1-1 proteins, respectively. In some observational studies of cancer, cruciferous vegetable intake confers greater protection in individuals with these polymorphisms; however, in other studies, the converse is observed. A recent study of sulforaphane pharmacokinetics suggests that lack of the GSTM1 enzyme is associated with more rapid excretion of sulforaphane. Many phytochemicals are also conjugated with glucuronide and sulfate moieties, and are excreted in urine and bile. Polymorphisms in UDP-glucuronosyltransferases (UGT) and sulfotransferases (SULT) may contribute to the variability in phytochemical clearance and efficacy. The effects of UGT polymorphisms on flavonoid clearance have not been examined, but UGT polymorphisms affect glucuronidation of several drugs and steroid hormones. Genetic polymorphisms in detoxification enzymes may account in part for individual variation in disease risk but have to be considered in the context of other aspects of human genetics, gut bacterial genetics, and environmental exposures. PMID- 17405688 TI - Xenoestrogens, biotransformation, and differential risks for breast cancer. PMID- 17405689 TI - Chemoprotection against cancer: an idea whose time has come. PMID- 17405690 TI - The role of mercury and cadmium heavy metals in vascular disease, hypertension, coronary heart disease, and myocardial infarction. AB - Mercury, cadmium, and other heavy metals have a high affinity for sulfhydryl ( SH) groups, inactivating numerous enzymatic reactions, amino acids, and sulfur containing antioxidants (NAC, ALA, GSH), with subsequent decreased oxidant defense and increased oxidative stress. Both bind to metallothionein and substitute for zinc, copper, and other trace metals reducing the effectiveness of metalloenzymes. Mercury induces mitochondrial dysfunction with reduction in ATP, depletion of glutathione, and increased lipid peroxidation; increased oxidative stress is common. Selenium antagonizes mercury toxicity. The overall vascular effects of mercury include oxidative stress, inflammation, thrombosis, vascular smooth muscle dysfunction, endothelial dysfunction, dyslipidemia, immune dysfunction, and mitochondrial dysfunction. The clinical consequences of mercury toxicity include hypertension, CHD, MI, increased carotid IMT and obstruction, CVA, generalized atherosclerosis, and renal dysfunction with proteinuria. Pathological, biochemical, and functional medicine correlations are significant and logical. Mercury diminishes the protective effect of fish and omega-3 fatty acids. Mercury, cadmium, and other heavy metals inactivate COMT, which increases serum and urinary epinephrine, norepinephrine, and dopamine. This effect will increase blood pressure and may be a clinical clue to heavy metal toxicity. Cadmium concentrates in the kidney, particularly inducing proteinuria and renal dysfunction; it is associated with hypertension, but less so with CHD. Renal cadmium reduces CYP4A11 and PPARs, which may be related to hypertension, sodium retention, glucose intolerance, dyslipidemia, and zinc deficiency. Dietary calcium may mitigate some of the toxicity of cadmium. Heavy metal toxicity, especially mercury and cadmium, should be evaluated in any patient with hypertension, CHD, or other vascular disease. Specific testing for acute and chronic toxicity and total body burden using hair, toenail, urine, serum, etc. with baseline and provoked evaluation should be done. PMID- 17405691 TI - Systems biology, toxins, obesity, and functional medicine. AB - By recognizing the role of toxins in obesity and altered function of the neuro endocrine-immune and mitochondrial and redox systems, and by creating a comprehensive strategy for both the reduction of exposure to and elimination of toxins, as well as the development of effective clinical strategies, treatment resistance in obesity may be more successfully addressed. Further research is needed to explore the clinical relevance and the mechanisms that underlie this hypothesis and to examine clinical detoxification methods. Through the prism of functional medicine, a context and road map exist for tackling many treatment resistant and complex chronic diseases, including obesity. PMID- 17405692 TI - Children's cognitive health: the influence of environmental chemical exposures. AB - The potential exists for developmental exposure of children to myriad chemicals, many of which are known to be neurotoxic. Some, such as the organophosphate pesticides, are specifically designed to attack the central nervous system. Despite the known and suspected risks associated with such exposures, critical aspects of the dose-response relationships are unknown or, at best, poorly characterized for the overwhelming majority of chemicals. Among the major knowledge gaps for most chemicals are the critical window(s) of vulnerability, the threshold or "no observed adverse effect level," and the host/environmental characteristics that modify individual vulnerability. Investigation of the role of genetic polymorphisms in determining vulnerability has barely begun. In the real-world, children are not exposed to a single chemical at a time but to complex mixtures of chemicals, and we have only a minimal understanding of the way in which exposures might interact with one another. Effective medical/environmental treatments for the adverse effects associated with chemical exposures are largely unknown, rendering primary prevention of exposure the most effective strategy for protecting children. PMID- 17405693 TI - Molds and mycotoxins: beyond allergies and asthma. PMID- 17405694 TI - Components of practical clinical detox programs--sauna as a therapeutic tool. AB - Saunas can be used very effectively for certain cardiovascular problems and as a means to enhance the mobilization of fat-soluble xenobiotics. When saunas are used to reduce blood pressure and enhance blood flow and cardiac functioning, only short sauna sessions (15 minutes) are necessary. When one wants to enhance the mobilization of heavy metals and chemical xenobiotics, longer sessions are needed and those should be medically monitored. But, for either use, saunas are safe and effective and should be used more frequently to benefit the health of our patients and ourselves. PMID- 17405695 TI - Toxin or medicine? Explanatory models of radon in Montana health mines. AB - Environmental protection and public health agencies in the United States and elsewhere label radioactive radon gas a toxic environmental hazard and a major cause of lung cancer. Paradoxically, in Europe and Japan radon gas is also used as an analgesic and anti-inflammatory, as one choice in the spectrum of conventional medical care. Although it is possible to find radon therapy in the United States, it exists only as an unconventional practice in Montana "radon health mines." In this article, I examine the use of radon therapy by Americans despite intensive public health education media campaigns. Using the notion of explanatory models as an analytical framework, I argue that American health mine clients adjust or replace "toxic models" of radon with new kinds of explanatory models that allow radon to be redefined as a healing substance. The manner of this adjustment varies according to peoples' individual needs, their own preexisting cultural models and experiences, and their individual personalities; the source of authoritative knowledge accepted by each person is a strong influence. Through these altered explanatory models, mine clients are able to view their use of radon therapy as a rational course of action. PMID- 17405696 TI - Perinatal sadness among Shuar women: support for an evolutionary theory of psychic pain. AB - Psychiatry faces an internal contradiction in that it regards mild sadness and low mood as normal emotions, yet when these emotions are directed toward a new infant, it regards them as abnormal. We apply parental investment theory, a widely used framework from evolutionary biology, to maternal perinatal emotions, arguing that negative emotions directed toward a new infant could serve an important evolved function. If so, then under some definitions of psychiatric disorder, these emotions are not disorders. We investigate the applicability of parental investment theory to maternal postpartum emotions among Shuar mothers. Shuar mothers' conceptions of perinatal sadness closely match predictions of parental investment theory. PMID- 17405697 TI - Herbalism, home gardens, and hybridization: Wothiha medicine and cultural change. AB - Using the example of the Wothiha of the Manapiare River Valley, Amazonas State, Venezuela, I challenge the image of the indigenous Amazonian as an expert in herbalism. I argue that the observed absence of medicinal plant use in early Wothiha ethnography, rather than reflecting researcher oversight, reflects the centrality of shamanism. According to Wothiha shamanic cosmology, herbal medicines, while useful to relieve symptoms and treat minor injuries, fail to address the underlying cause of illness. Using a combination of quantitative and qualitative methods, I find that as the role and influence of shamanism have dramatically decreased, the Wothiha have turned elsewhere for medical treatment. Biomedical remedies have shown to be effective, thereby encouraging an acceptance of symptom-specific treatments. Biomedicine's patchy availability, however, has encouraged the Wothiha to look beyond biomedicine. Several folk healing traditions are being incorporated by the Wothiha, each with its own herbal tradition. PMID- 17405698 TI - Maternal bodies, breast-feeding, and consumer desire in urban China. AB - Urban Chinese women in the 1990s formulated their infant-feeding decisions in the context of a society undergoing radical transformation as the nation moved from a centrally planned socialist economy to a global, market-oriented one. Narratives of new mothers in Beijing in the 1990s provide insights into the multiple forces that shaped their infant-feeding practices. These personal histories also illustrate the limitations of multilateral breast-feeding programs that emphasize breast-feeding as a natural interaction between mother and infant. The cases I present here demonstrate instead that the material, bodily manifestations of breast-feeding require nursing mothers to continually renegotiate relations with husbands, coworkers, and family. Chinese women's accounts also add insight to theoretical deliberations on gender and the body, for they demonstrate that cultural expectations and the demands of the lactating body must be considered to understand fully the process of women's decisions in a social and not strictly reproductive context. On a larger scale, the data also illustrate how global intervention, in the form of the WHO-UNICEF-sponsored Baby-Friendly Hospital Initiative, promotes breast-feeding as a woman's primary duty at the same time that market forces counter this message as women redefine their individual expectations and social relationships. PMID- 17405699 TI - Orthostatic panic as a key Vietnamese reaction to traumatic events: the case of September 11, 2001. AB - This article discusses a culturally specific response to traumatic events: orthostatic panic attacks among Vietnamese refugees. We compared the rate and severity of orthostatic panic as well as the rates and severity of associated flashbacks a month before and a month after September 11, 2001. After that date, the rate and severity of orthostatic panic greatly increased, as did the rate and severity of associated flashbacks. The central role of orthostatic panic as a response to traumatic events is illustrated through a patient's vignette. An explanation of why September 11 so profoundly influenced this population is adduced, including an explanation of why it resulted in considerable worsening of orthostatic panic. PMID- 17405700 TI - Durbolota (weakness), chinta rog (worry illness), and poverty: explanations of white discharge among married adolescent women in an urban slum in Dhaka, Bangladesh. AB - I carried out ethnographic fieldwork among 153 married adolescent girls, aged 15 19, in a Dhaka slum from December 2001 to January 2003, including 50 in-depth interviews and eight case studies. I also held discussions with family and community members. In this article, I focus on popular understandings of vaginal discharge being caused by durbolota (weakness) and chinta rog (worry illness), as mentioned by young women. Eighty-eight young women reported that they had experienced white discharge, blaming it on a number of factors such as stress and financial hardships, tensions in the household, marital instability, hunger anxiety, and reproductive burdens. For married adolescent women in the urban slum, white discharge has many levels of meaning linked to the broader social, political, and material inequalities in their everyday lives. PMID- 17405701 TI - [Not Available]. PMID- 17405703 TI - [Not Available]. PMID- 17405702 TI - [Not Available]. PMID- 17405704 TI - [Not Available]. PMID- 17405705 TI - [Not Available]. PMID- 17405706 TI - [Not Available]. PMID- 17405707 TI - [Not Available]. PMID- 17405708 TI - [Not Available]. PMID- 17405709 TI - [Not Available]. PMID- 17405710 TI - [Not Available]. PMID- 17405711 TI - [Not Available]. PMID- 17405712 TI - [Not Available]. PMID- 17405713 TI - [Not Available]. PMID- 17405714 TI - [Not Available]. PMID- 17405715 TI - [Not Available]. PMID- 17405716 TI - [Not Available]. PMID- 17405717 TI - [Not Available]. PMID- 17405718 TI - [Not Available]. PMID- 17405720 TI - [Not Available]. PMID- 17405719 TI - [Not Available]. PMID- 17405721 TI - [Not Available]. PMID- 17405722 TI - [Not Available]. PMID- 17405724 TI - [Not Available]. PMID- 17405723 TI - [Not Available]. PMID- 17405725 TI - [Not Available]. PMID- 17405726 TI - [Not Available]. PMID- 17405727 TI - [Not Available]. PMID- 17405728 TI - [Not Available]. PMID- 17405729 TI - [Not Available]. PMID- 17405730 TI - [Not Available]. PMID- 17405731 TI - [Not Available]. PMID- 17405732 TI - [Not Available]. PMID- 17405734 TI - [Not Available]. PMID- 17405733 TI - [Not Available]. PMID- 17405735 TI - [Not Available]. PMID- 17405736 TI - [Not Available]. PMID- 17405738 TI - [Not Available]. PMID- 17405737 TI - [Not Available]. PMID- 17405739 TI - The bHLH transcription factor Tcf12 (ME1) mRNA is abundantly expressed in Paneth cells of mouse intestine. AB - Using a large-scale in situ hybridization screening system, we found that mRNA coding for ME1, a basic helix-loop-helix (bHLH) transcription factor, was abundantly expressed in Paneth cells of adult small intestinal crypts. Other functionally related E-protein mRNAs, ME2, and E2A, however, could not be detected in the cells. ME1 mRNA was first detected in the jejunum and ileum two weeks after birth when the number of Paneth cells starts to increase. ME1 is the first identified bHLH transcription factor expressed in the Paneth cells and may be used as a molecular marker and a key molecule for analyzing transcriptional regulation in the Paneth cell. PMID- 17405740 TI - Importance of allogeneic T-cells for disease control after stem cell transplantation for high-risk Langerhans cell histiocytosis. AB - Reduced intensity conditioning followed by allogeneic SCT (RIC-SCT) has recently emerged as promising new salvage option for children suffering from Langerhans cell histiocytosis (LCH) with risk organ involvement and failure to conventional therapy. We report on the posttransplant course of female toddler with high-risk LCH, who achieved complete remission after RIC-SCT, despite a posttransplant chimerism constellation, in which only the T-cell subset proved to be of donor origin in the long-term. We therefore suggest that allogeneic T-cells have played a crucial role in controlling disease activity in this patient and may exert the major curative effect after RIC-SCT for LCH. PMID- 17405741 TI - Cutaneous Mycobacterium chelonae infection in chronic lymphocytic leukaemia. PMID- 17405742 TI - Successful treatment with T-depleted autologous peripheral blood stem cell transplantation of refractory chronic autoimmune thrombocytopenic purpura. AB - Autoimmune thrombocytopenia (AITP) is a disorder due to specific platelet auto antibodies directed against platelet surface glycoproteins. AITP in adults is usually chronic, idiopathic and frequently refractory to conventional treatments. Myelo- and immuno-suppressive chemotherapy followed by autologous peripheral blood stem cell (PBSC) transplantation is an experimental approach for severe chronic refractory AITP. We report a case of a woman with AITP, refractory to the conventional therapy, submitted to T-cell-depleted autologous PBSC transplantation, which obtained long term stable response on platelet count. We deem that the positive outcome of our patient depends on T-cells depletion of the graft, which reduces autoreactive T clones. PMID- 17405743 TI - Two successful pregnancies in a chronic myeloid leukemia patient treated with imatinib. AB - The number of CML patients in child-bearing age and treated with imatinib is increasing. These women may want to be pregnant or are actually pregnant while on imatinib. Physicians do not know when to stop the treatment and what the risks are for the foetus and the mother. We report a case of a CML patient treated with imatinib who has two children, now 3 years and 10 months of age, in good health. The mother was in complete molecular remission, relapsed during pregnancy and reverted to remission in both cases after delivery. PMID- 17405744 TI - AIDS-related plasmablastic lymphoma of the oral cavity associated with an IGH/MYC translocation--treatment with autologous stem-cell transplantation in a patient with severe haemophilia-A. AB - Plasmablastic lymphoma is an AIDS related lymphoma that continues to have a poor prognosis despite significant advances in the management of HIV and lymphoproliferative diseases. In part this has been due to limited insights into the biology of this disease and the molecular mechanisms of oncogenesis. To date molecular abnormalities have not been described in plasmablastic lymphoma, and its aggressive clinical behaviour has been difficult to understand. We describe the first reported cytogenetic abnormality in plasmablastic lymphoma, an IgH/MYC translocation. It is also the first description of autologous stem cell transplantation in a patient with severe haemophilia A. PMID- 17405745 TI - Anaemia and abdominal pain due to occupational lead poisoning. AB - We describe a 47-year-old patient with chronic anaemia with basophilic stippling of erythrocytes, recurrent abdominal colics, discoloration of gums, sensitive polyneuropathy to the four limbs, hyperuricaemia, hepatosteatosis with raised transaminases, and a long ignored history of lead exposure in a battery recycling plant. The diagnosis of poisoning was confirmed by high lead levels in the blood and urine, decreased erythrocyte delta-aminolevulinic acid dehydratase (ALA-D), raised erythrocyte zinc protoporphyrin (ZP), and elevated urinary excretion of porphyrins. Chelation with EDTA resulted in increased urinary lead excretion, gradual improvement of the clinical picture, and progressive normalization of lead biomarkers. The case highlights the importance of occupational anamnesis for the diagnosis of lead poisoning, an uncommon condition which may mimic a variety of internal and surgical diseases. Since antiquity, lead has been extensively mined, produced, and utilized in a variety of industrial settings, such as metallurgy, construction, production of plastics, ceramics, paints and pigments. Lead and its compounds are systemic toxicants, and a wide range of adverse health effects (including haematological, gastrointestinal, neuropsychiatric, cardiovascular, renal, endocrine, and reproductive disorders) has been observed in exposed workers. The general population (particularly children) may also be exposed to toxic lead levels due to air, soil, food and water contamination. Thanks to the improvement of workplace hygienic conditions, the pathological picture of occupational lead poisoning (plumbism, saturnism) has gradually become less serious, at least in the most industrialized countries, and has progressively changed into aspecific, subclinical manifestations. We describe here an unusual case (nowadays) of anaemia and recurrent abdominal pain due to lead poisoning from battery recycling. PMID- 17405746 TI - Mini-dose of thalidomide for treatment of primary myelofibrosis. Report of a case with complete reversal of bone marrow fibrosis and splenomegaly. AB - We report a patient with very advanced myelofibrosis and huge splenomegaly who showed a complete hematological response to low dose thalidomide with reversal of splenomegaly and bone narrow fibrosis after 30 months of the treatment. PMID- 17405747 TI - An acquired inhibitor that produced a delay of fibrinopeptide B release in an asymptomatic patient. AB - An asymptomatic, 29-year-old woman was referred to our hospital before surgery because in the basic study of hemostasis she showed a prolonged thrombin time (TT) and a normal reptilase time (RT). She had not received any anticoagulants so, to account for these abnormal results the presence of an inhibitor or a dysfibrinogenemia was suspected. A 1:1 mixture of the patient's plasma with control plasma did not correct the TT. Dysfibrinogenemia was excluded because the defibrinated plasma retained the inhibitory activity when mixed with normal plasma. When 0.02 mg/ml of Protamine Sulphate (a concentration that neutralizes 1 U/mL of heparin in normal plasma) was added to the patient's plasma, the inhibitory activity did not disappear. IgG from the patient and from normal serum was isolated. The patient's IgG was able to prolong the TT of a normal plasma and of a purified fibrinogen. The patient IgG did not impair the catalytic activity of thrombin, because no difference was observed in the hydrolysis of S-2238 by 1 U NIH human thrombin with normal or patient IgG. The time course of the thrombin mediated fibrinopeptide-release from normal fibrinogen with the patient's IgG, showed a delay in the fibrinopeptide B (FPB) release without affecting the fibrinopeptide A (FPA) release. This patient has an IgG antibody that delays fibrinopeptide B release of fibrinogen. PMID- 17405748 TI - Re-occurrence of the CD20 molecule expression subsequent to CD20-negative relapse in diffuse large B-cell lymphoma. AB - We report the first case of diffuse large B-cell lymphoma (DLBCL) of the stomach displaying CD20-negative relapse after rituximab-containing treatment and the re appearance of CD20 expression at the second failure. The loss of CD20 expression in B-cell lymphomas relapsing after rituximab is a well-known phenomenon, but its actual impact in DLBCL is difficult to estimate. This paradigmatic case suggests that CD20-expression reappearance after purging of CD20-positive clones with rituximab might be an underestimated occurrence in B-cell lymphomas. Accordingly, every relapse, whenever possible, should be histologically assessed with diagnostic and immunophenotyping purposes. PMID- 17405749 TI - Late-onset neutropenia following rituximab results from a hematopoietic lineage competition due to an excessive BAFF-induced B-cell recovery. AB - Rituximab is used in the treatment of lymphoma and autoimmune diseases, for which late-onset neutropenia (LON) were reported. LON-related mechanisms remain unclear. To obtain insights into the mechanisms, we assessed serum, peripheral blood and bone marrow (BM) samples of a patient with LON. Factors classically associated with neutropenia such as anti-neutrophil antibodies, T-LGL, soluble Fas Ligand were not detectable. We then evaluated the kinetics of various cytokines involved in B-cell and granulocyte homeostasis. We found that LON is related to a lack of granulopoiesis in the BM that coincides with a very high level of BAFF, a strong stimulator of B-cell recovery, and hypothesized a hematopoietic lineage competition due to an excessive B-cell recovery in the BM by promotion of B-cell lymphopoiesis over granulopoiesis within common developmental niches. Assessment of serum BAFF levels following rituximab could detect patients at risk of developing LON. PMID- 17405750 TI - Flow cytometry characterization of leukemic phase of nasal NK/T-cell lymphoma in tumor biopsies and peripheral blood. AB - We report the findings of the immunophenotypic profile of three cases of nasal T/NK cell lymphoma in leukemic phase. Flow cytometry analysis was carried out using cell suspensions of tumor nasal biopsies and peripheral blood. Tumor samples were composed by a mixture of a predominant subset of medium-size true NK cytCD3epsilon-, sCD3epsilon-, CD56+ cells mixed with a minor subset of medium size T/NK sCD3epsilon+, CD56+ cells. Both subsets were also detected in peripheral blood. In addition, an infiltration of small-size sCD3epsilon+, CD56- normal T lymphocytes was also present. PMID- 17405751 TI - Successful treatment of a primary uterine B-cell lymphoma with rituximab-CHOP immunochemotherapy. AB - We report the case of a 26 year old patient with a primary, Ann Arbor stage IE, diffuse large B-cell lymphoma of the uterine corpus. The patient was admitted to our hospital because of a one year history of vaginal contact bleeding. Final diagnosis was based on the histological and immunohistochemical evaluation of a specimen obtained by fractionated cervical and uterine curettage. Further staging has not revealed any other localization of the disease. Antilymphoma treatment was started with CHOP combined with rituximab and resulted complete remission. This is the first reported case of an uterine lymphoma treated by rituximab-based immunochemotherapy. PMID- 17405752 TI - Paraplegia due to extramedullary hematopoiesis in thalassemia treated successfully with radiation therapy. AB - Spinal cord compression due to extramedullary hematopoiesis (EMH) is a rare complication of thalassemia and generally presents as paraparesis with sensory impairment. Complete paraplegia is extremely rare in EMH due to thalassemia although it is known to occur in polycythemia vera and sickle cell anemia. Treatment options mostly include surgery and/or radiotherapy. Whereas cases presenting with paraparesis have been treated with either surgery or radiotherapy with equal frequency and efficacy, almost all reported cases with paraplegia have been treated with surgery with or without radiation therapy. We hereby report a case of thalassemia intermedia with paraplegia treated successfully with radiotherapy. PMID- 17405753 TI - Erythopoietin treatment during complement inhibition with eculizumab in a patient with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is characterized by intravascular hemolysis leading to anemia and other clinical manifestations. Transfusions are often required to support hemoglobin at tolerable levels. A PNH patient with aplastic anemia was treated with the complement inhibitor eculizumab, followed by concurrent treatment with recombinant human erythropoietin (rHuEpo). Eculizumab alone reduced hemolysis, increased PNH red blood cell (RBC) mass, and decreased transfusions. Addition of rHuEpo during eculizumab therapy, enhanced erythropoiesis, further increased PNH RBC mass and hemoglobin levels, and rendered the patient transfusion independent for more than two years. These data show that driving erythropoiesis during eculizumab treatment provided further benefit to a patient with PNH and underlying bone marrow failure. PMID- 17405754 TI - Clinically and genetically atypical T-cell prolymphocytic leukemia underlines the relevance of a multidisciplinary diagnostic approach. PMID- 17405755 TI - Primary T-cell lymphoma of the thyroid gland with chemokine receptors of Th1 phenotype complicating autoimmune thyroiditis. AB - Lymphoma of the thyroid is almost exclusively derived from B cells of mucosa associated lymphoid tissue (MALT), and frequently co-exist with autoimmune thyroiditis in which most infiltrating cells are of Th1 cell origin. We present here two rare cases of peripheral T-cell lymphoma (PTCL) based on chronic thyroiditis with the phenotype CD3+, CD4+, CD8-, TCR+. Furthermore, lymphoma cells in both cases were CXCR3+, CCR5+ and ST2(L)-, suggesting a Th1 cell origin. Eight of 11 cases of PTCL of the thyroid in the literature, including our cases, were associated with thyroiditis. Except for one tumor of T-cell type, all of the five lymphomas analyzed for CD4 expression were positive for the antigen. Among them, both those examined for chemokine receptors were phenotypically of Th1-cell origin with a background of thyroiditis, suggesting that Th1 activation induced by chronic inflammation could lead to PTCL of themselves as well as MALT-lymphoma of B cells. PMID- 17405756 TI - Cocaine abuse may influence the response to imatinib in CML patients. PMID- 17405757 TI - Large granular lymphocyte proliferation and revertant mosaicism: two rare events in a Wiskott-Aldrich syndrome patient. AB - We report on a 6 year old patient with an unusual clinical presentation of WAS and oligoclonal proliferation of TCR+ large granular lymphocytes (LGL). Flow cytometry demonstrated two distinct populations of lymphocytes with strongly decreased (WASP-) or normal expression levels of WASP (WASP+), respectively. Molecular analysis confirmed a splice site mutation in intron 2 of the WASP gene in the WASP- cells but not in WASP+ cells. LGL cells were WASP+, suggesting that two independent rare events, somatic revertant mosaicism and LGL expansion, have occurred in a child with WAS. Our report points to diagnostic difficulties in the presence of partial WASP reversions and LGL. PMID- 17405758 TI - RE 10104: Abou-Jawde et al. The role of race, socioeconomic status, and distance traveled on the outcome of African-American patients with multiple myeloma. Haematologica 2006; 91: 1410-1413. PMID- 17405759 TI - Marrow fibrosis does not account for circulating CD34+ cells in myelofibrosis with myeloid metaplasia. PMID- 17405760 TI - Lipoprotein(a), athero-thrombosis and longevity. A historical paradox finally elucidated? PMID- 17405761 TI - Most people over age 50 in the general population do not meet ACR remission criteria or OMERACT minimal disease activity criteria for rheumatoid arthritis. AB - OBJECTIVE: To analyse the proportion of individuals in the general population over age 50 who do not meet American College of Rheumatology (ACR) criteria for rheumatoid arthritis (RA) remission, and OMERACT criteria for minimal disease activity (MDA), and to compare results to RA patients. METHODS: A self-report questionnaire was completed by 1400 community control subjects and 1705 RA patients, including the Health Assessment Questionnaire (HAQ), gradual rating scales for pain, fatigue and global health, duration of morning stiffness and painful joints. The prevalence of 4/6 ACR remission criteria and 4/7 OMERACT criteria for MDA was analysed in community control subjects and patients with RA over age 50. RESULTS: For ACR criteria, 76% of control subjects reported painful joints, 37% morning stiffness, 62% pain and 66% fatigue, vs 94, 65, 84 and 84% of patients with RA. MDA criteria were not met by 64% of control subjects for painful joints, 38% for pain, 45% for global health and 18% for HAQ, vs 89, 60, 69 and 52% of RA patients. The four ACR remission criteria were met by only 15% of control subjects over age 50 and 3% of RA patients, and MDA criteria by 28% of controls and 7% of patients. CONCLUSIONS: The majority of community population over age 50 did not meet criteria for remission or MDA in RA. Although a self report format may differ from results involving an assessor, the current criteria may not be accurate to identify remission or MDA in people with RA who are older than age 50. PMID- 17405762 TI - Loss of aquaporin 4 in lesions of neuromyelitis optica: distinction from multiple sclerosis. AB - Neuromyelitis optica (NMO) is an inflammatory and necrotizing disease clinically characterized by selective involvement of the optic nerves and spinal cord. There has been a long controversy as to whether NMO is a variant of multiple sclerosis (MS) or a distinct disease. Recently, an NMO-specific antibody (NMO-IgG) was found in the sera from patients with NMO, and its target antigen was identified as aquaporin 4 (AQP4) water channel protein, mainly expressed in astroglial foot processes. However, the pathogenetic role of the AQP4 in NMO remains unknown. We did an immunohistopathological study on the distribution of AQP4, glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), activated complement C9neo and immunoglobulins in the spinal cord lesions and medulla oblongata of NMO (n = 12), MS (n = 6), brain and spinal infarction (n = 7) and normal control (n = 8). The most striking finding was that AQP4 immunoreactivity was lost in 60 out of a total of 67 acute and chronic NMO lesions (90%), but not in MS plaques. The extensive loss of AQP4 accompanied by decreased GFAP staining was evident, especially in the active perivascular lesions, where immunoglobulins and activated complements were deposited. Interestingly, in those NMO lesions, MBP-stained myelinated fibres were relatively preserved despite the loss of AQP4 and GFAP staining. The areas surrounding the lesions in NMO had enhanced expression of AQP4 and GFAP, which reflected reactive gliosis. In contrast, AQP4 immunoreactivity was well preserved and rather strongly stained in the demyelinating MS plaques, and infarcts were also stained for AQP4 from the very acute phase of necrosis to the chronic stage of astrogliosis. In normal controls, AQP4 was diffusely expressed in the entire tissue sections, but the staining in the spinal cord was stronger in the central grey matter than in the white matter. The present study demonstrated that the immunoreactivities of AQP4 and GFAP were consistently lost from the early stage of the lesions in NMO, notably in the perivascular regions with complement and immunoglobulin deposition. These features in NMO were distinct from those of MS and infarction as well as normal controls, and suggest that astrocytic impairment associated with the loss of AQP4 and humoral immunity may be important in the pathogenesis of NMO lesions. PMID- 17405763 TI - Paul Broca's historic cases: high resolution MR imaging of the brains of Leborgne and Lelong. AB - In 1861, the French surgeon, Pierre Paul Broca, described two patients who had lost the ability to speak after injury to the posterior inferior frontal gyrus of the brain. Since that time, an infinite number of clinical and functional imaging studies have relied on this brain-behaviour relationship as their anchor for the localization of speech functions. Clinical studies of Broca's aphasia often assume that the deficits in these patients are due entirely to dysfunction in Broca's area, thereby attributing all aspects of the disorder to this one brain region. Moreover, functional imaging studies often rely on activation in Broca's area as verification that tasks have successfully tapped speech centres. Despite these strong assumptions, the range of locations ascribed to Broca's area varies broadly across studies. In addition, recent findings with language-impaired patients have suggested that other regions also play a role in speech production, some of which are medial to the area originally described by Broca on the lateral surface of the brain. Given the historical significance of Broca's original patients and the increasing reliance on Broca's area as a major speech centre, we thought it important to re-inspect these brains to determine the precise location of their lesions as well as other possible areas of damage. Here we describe the results of high resolution magnetic resonance imaging of the preserved brains of Broca's two historic patients. We found that both patients' lesions extended significantly into medial regions of the brain, in addition to the surface lesions observed by Broca. Results also indicate inconsistencies between the area originally identified by Broca and what is now called Broca's area, a finding with significant ramifications for both lesion and functional neuroimaging studies of this well-known brain area. PMID- 17405764 TI - Novel locus for benign hereditary chorea with adult onset maps to chromosome 8q21.3 q23.3. AB - Autosomal dominant choreas are genetically heterogeneous disorders including Huntington disease (HD), Huntington disease like 1 (HDL1), Huntington disease like 2 (HDL2), dentatorubro-pallidoluysian atrophy (DRPLA), spinocerebellar ataxia type 17 (SCA17) and benign hereditary chorea (BHC). We identified two Japanese families with adult-onset benign chorea without dementia inherited in an autosomal dominant pattern. All affected individuals presented slowly progressive choreic movements in their upper and lower extremities, trunk and head with an age of onset ranging from 40 to 66 (average 54.3), which were markedly improved by haloperidol. The affected individuals also developed reduced muscle tones in their extremities. The findings obtained in the brain CT or MRI studies of nine affected individuals were normal. These clinical features resemble those of the so-called 'senile chorea'. HD, HDL1, HDL2, DRPLA, SCA17 and BHC caused by mutations in the TITF-1 gene were excluded by mutational and linkage analyses. A genome-wide linkage analysis revealed linkage to chromosome 8q21.3-q23.3 with a maximum cumulative two-point log of the odds (LOD) score of 4.74 at D8S1784 (theta = 0.00). Haplotype analysis of both the families defined the candidate region as 21.5 Mb interval flanked by M9267 and D8S1139. We named this adult onset dominant inherited chorea 'benign hereditary chorea type 2 (BHC2)'. PMID- 17405765 TI - Locomotor adaptation on a split-belt treadmill can improve walking symmetry post stroke. AB - Human locomotion must be flexible in order to meet varied environmental demands. Alterations to the gait pattern occur on different time scales, ranging from fast, reactive adjustments to slower, more persistent adaptations. A recent study in humans demonstrated that the cerebellum plays a key role in slower walking adaptations in interlimb coordination during split-belt treadmill walking, but not fast reactive changes. It is not known whether cerebral structures are also important in these processes, though some studies of cats have suggested that they are not. We used a split-belt treadmill walking task to test whether cerebral damage from stroke impairs either type of flexibility. Thirteen individuals who had sustained a single stroke more than 6 months prior to the study (four females) and 13 age- and gender-matched healthy control subjects were recruited to participate in the study. Results showed that stroke involving cerebral structures did not impair either reactive or adaptive abilities and did not disrupt storage of new interlimb relationships (i.e. after-effects). This suggests that cerebellar interactions with brainstem, rather than cerebral structures, comprise the critical circuit for this type of interlimb control. Furthermore, the after-effects from a 15-min adaptation session could temporarily induce symmetry in subjects who demonstrated baseline asymmetry of spatiotemporal gait parameters. In order to re-establish symmetric walking, the choice of which leg is on the fast belt during split-belt walking must be based on the subject's initial asymmetry. These findings demonstrate that cerebral stroke survivors are indeed able to adapt interlimb coordination. This raises the possibility that asymmetric walking patterns post-stroke could be remediated utilizing the split belt treadmill as a long-term rehabilitation strategy. PMID- 17405766 TI - Viral vector-induced amygdala NPY overexpression reverses increased alcohol intake caused by repeated deprivations in Wistar rats. AB - Acute administration of neuropeptide Y (NPY) modulates alcohol intake in genetic and chemical models of high intake, while leaving intake unaffected during 'normal' or baseline conditions. In non-selected, normal rat lines, alcohol consumption can be increased by prolonged exposure to alcohol, and it is unclear what effect a constitutive increase in NPY function will have on alcohol intake. The purpose of the present study was to examine the effects on alcohol intake of an inducible, constitutive overexpression of NPY, one of the most abundant neuropeptides in the central nervous system. A liquid diet was used in combination with repeated alcohol deprivation sessions to increase alcohol intake in normal Wistar rats. We then examined the effect of NPY overexpression in the amygdala on excessive alcohol intake produced by prolonged exposure to alcohol and alcohol deprivation. Repeated withdrawal increased alcohol consumption in a 24-h continuous access two-bottle choice model. Both the number of withdrawals as well as the length of the withdrawal periods affected alcohol consumption with an increased intake resulting from multiple withdrawals and the alcohol deprivation effect being enhanced by longer periods of abstinence. The increase in intake following repeated abstinence was blunted by intra-amygdala administration of a Sindbis viral vector containing NPY cDNA. Amygdala NPY overexpression also was demonstrated to be anxiolytic in the open field test. Repeated withdrawal in combination with a history of alcohol consumption significantly elevated alcohol intake, and the amygdala may mediate the transition to high-drinking states in this model. PMID- 17405767 TI - A clinical rating scale for progressive supranuclear palsy. AB - We devised a Progressive Supranuclear Palsy (PSP) Rating Scale comprising 28 items in six categories: daily activities (by history), behaviour, bulbar, ocular motor, limb motor and gait/midline. Scores range from 0 to 100, each item graded 0-2 (six items) or 0-4 (22 items). Inter-rater reliability is good, with intra class correlation coefficient for the overall scale of 0.86 (95% CI 0.65-0.98). A single examiner applied the PSPRS at every visit for 162 patients. Mean rate of progression was 11.3 (+/-11.0) points per year. Neither onset age nor gender correlated well with rate of progression. Median actuarially corrected survival was 7.3 years. The PSPRS score was a good independent predictor of subsequent survival (P < 0.0001). For example, for patients with scores from 40 to 49, 3 year survival was 41.9% (95% CI 31.0-56.6) but 4-year survival was only 17.9% (95% CI 10.2-31.5). For those patients, likelihood or retaining some gait function was 51.7% (40.0-66.9) at 1 year but only 6.5% (1.8-23.5) at 3 years. We conclude that the PSPRS is a practical measure that is sensitive to disease progression and could be useful as a dependent variable in observational or interventional trials and as an indicator of prognosis in clinical practice. PMID- 17405768 TI - Parallel gene synthesis in a microfluidic device. AB - The ability to synthesize custom de novo DNA constructs rapidly, accurately and inexpensively is highly desired by researchers, as synthetic genes and longer DNA constructs are enabling to numerous powerful applications in both traditional molecular biology and the emerging field of synthetic biology. However, the current cost of de novo synthesis-driven largely by reagent and handling costs-is a significant barrier to the widespread availability of such technology. In this work, we demonstrate, to our knowledge, the first gene synthesis in a microfluidic environment. The use of microfluidic technology greatly reduces reaction volumes and the corresponding reagent and handling costs. Additionally, microfluidic technology enables large numbers of complex reactions to be performed in parallel. Here, we report the fabrication of a multi-chamber microfluidic device and its use in carrying out the syntheses of several DNA constructs. Genes up to 1 kb in length were synthesized in parallel at minute starting oligonucleotide concentrations (10-25 nM) in four 500 nl reactors. Such volumes are one to two orders of magnitude lower than those utilized in conventional gene synthesis. The identity of all target genes was verified by sequencing, and the resultant error rate was determined to be 1 per 560 bases. PMID- 17405769 TI - Carbodiimide-mediated cross-linking of RNA to nylon membranes improves the detection of siRNA, miRNA and piRNA by northern blot. AB - The northern blot, or RNA gel blot, is a widely used method for the discovery, validation and expression analysis of small regulatory RNA such as small interfering RNA (siRNA), microRNA (miRNA) and piwi-interacting RNA (piRNA). Although it is straightforward and quantitative, the main disadvantage of a northern blot is that it detects such RNA less sensitively than most other approaches. We found that the standard dose of UV used in northern blots was not the most efficient at immobilizing small RNA of 20-40 nt on nylon membranes. However, increasing the dose of UV reduced the detection of miRNA by hybridization in northern blotting experiments. We discovered that using the soluble carbodiimide, EDC, to cross-link RNA to nylon membranes greatly improved the detection of small RNA by hybridization. Compared to standard UV cross linking procedures, EDC cross-linking provided a 25-50-fold increase in the sensitivity of detection of siRNA from plants and miRNA or piRNA from mammalian cells. All types of hybridization probes tested benefited from the new cross linking procedure. Cross-linking was dependent on a terminal phosphate and so, should be applicable to other related categories of small RNA. PMID- 17405770 TI - Treatment and outcomes of patients with evolving myocardial infarction: experiences from the SYNERGY trial. AB - AIMS: Patients with myocardial infarction (MI) presenting immediately after symptom onset may be treated less aggressively due to their non-elevated troponin status. We compared the initial treatment and clinical outcomes of patients presenting with evolving MI (EMI) with those presenting with MI. METHODS AND RESULTS: This study analysed data from the Superior Yield of the New strategy of Enoxaparin, Revascularisation, and Glycoprotein IIb/IIIa inhibitors (SYNERGY) trial, which enrolled patients meeting at least two of the following: age >or= 60 years, elevated cardiac biomarkers, or ST-segment changes. Patients were stratified by troponin results obtained within 12 h of presentation: EMI [initial troponin (-), second troponin (+)], MI [initial troponin (+)], and no MI at enrolment [first and second troponin (-)]. Comparisons were made using Wilcoxon rank-sum and chi(2) tests. Of the 8,309 patients with complete data, 5,503 (66%) had MI, 1,686 (20%) had EMI, and 1,120 (13%) had no MI. Treatment patterns prior to enrolment were similar among EMI and MI patients [aspirin (88 vs. 86%), beta blockers (62 vs. 61%), heparin (83 vs. 81%), and glycoprotein IIb/IIIa inhibitors (23 vs. 24%)]. Similar rates of percutaneous coronary intervention (48 vs. 50%) and coronary artery bypass grafting (21 vs. 22%) were seen after enrolment. Patients presenting with MI had a higher rate of death or recurrent MI compared with patients with EMI [16 vs. 13%, adjusted OR 1.22 (95% CI 1.04, 1.44)]. CONCLUSION: Initial treatment patterns were similar among patients with EMI and MI in the SYNERGY trial. Patients with EMI had lower rates of death or re infarction at 30 days compared with patients presenting with positive troponin results. PMID- 17405771 TI - Enzymatic synthesis of cello-oligosaccharides by rice BGlu1 {beta}-glucosidase glycosynthase mutants. AB - Rice BGlu1 beta-glucosidase is a glycosyl hydrolase family 1 enzyme that acts as an exoglucanase on beta-(1,4)- and short beta-(1,3)-linked gluco oligosaccharides. Mutations of BGlu1 beta-glucosidase at glutamate residue 414 of its natural precursor destroyed the enzyme's catalytic activity, but the enzyme could be rescued in the presence of the anionic nucleophiles such as formate and azide, which verifies that this residue is the catalytic nucleophile. The catalytic activities of three candidate mutants, E414G, E414S, and E414A, in the presence of the nucleophiles were compared. The E414G mutant had approximately 25 and 1400-fold higher catalytic efficiency than E414A and E414S, respectively. All three mutants could catalyze the synthesis of mixed length oligosaccharides by transglucosylation, when alpha-glucosyl fluoride was used as donor and pNP cellobioside as acceptor. The E414G mutant gave the fastest transglucosylation rate, which was approximately 3- and 19-fold faster than that of E414S and E414A, respectively, and gave yields of up to 70-80% insoluble products with a donor acceptor ratio of 5:1. (13)C-NMR, methylation analysis, and electrospray ionization-mass spectrometry showed that the insoluble products were beta-(1,4) linked oligomers with a degree of polymerization of 5 to at least 11. The BGlu1 E414G glycosynthase was found to prefer longer chain length oligosaccharides that occupy at least three sugar residue-binding subsites as acceptors for productive transglucosylation. This is the first report of a beta-glucansynthase derived from an exoglycosidase that can produce long-chain cello-oligosaccharides, which likely reflects the extended oligosaccharide-binding site of rice BGlu1 beta glucosidase. PMID- 17405772 TI - PTEN regulates phospholipase D and phospholipase C. AB - PTEN is an ubiquitously expressed tumor suppressor which plays a prominent role in the pathogenesis of many types of sporadic solid tumors, including breast cancer, as well as hematologic malignancies. Germline PTEN mutations cause 85% of Cowden syndrome (CS), characterized by a high risk of breast and thyroid cancers, and 65% of Bannayan-Riley-Ruvalcaba syndrome (BRRS), characterized by lipomatosis, hemangiomas and speckled penis. Historically, PTEN's role in tumor suppression has been linked to the down-regulation of the PI3K/AKT pathway by PTEN's lipid phosphatase activity. Beyond the AKT pathway, however, there has been minimal examination of PTEN's responsibility in lipid-derived cellular signaling. As phospholipids have been shown to be critical components in signal transduction and cellular proliferation and PTEN controls cellular phospholipid levels, we hypothesized that PTEN functions as a regulator of lipid signaling and homeostasis. Increased PTEN expression in unstimulated MCF-7 breast cancer cells results in a 51% increase in phosphatidic acid, with a decrease in phosphatidylcholine, suggesting that PTEN may regulate phospholipase D (PLD). PTEN overexpression results in a 30% increase in basal PLD activity. As phospholipase C (PLC) is both involved in PLD activation and is regulated by PIP2/3 levels, we investigated the role of PTEN on PLC activation. Our data suggest that PTEN modulates PLC:PLD activation pathways and indicate that the pathogenesis of CS/BRRS has a more complex biochemical basis beyond simply activating the PI3K pathway. This provides alternative routes for PTEN's tumor suppressor action that may be beneficial in the creation of novel targets for cancer therapy and prevention. PMID- 17405773 TI - Physicians' experiences using commercial e-prescribing systems. AB - Public and private efforts are under way to promote electronic prescribing to improve health care safety, quality, and efficiency. Findings from this qualitative study of physician practices suggest that substantial gaps may exist between advocates' vision of e-prescribing and how physicians use commercial e prescribing systems today. While physicians were positive about the most basic e prescribing features, they reported major barriers to maintaining complete patient medication lists, using clinical decision support, obtaining formulary data, and electronically transmitting prescriptions to pharmacies. Three factors help explain the gaps: product limitations, external implementation challenges, and physicians' preferences about using specific product features. PMID- 17405774 TI - NICE guidelines on urinary incontinence in women. PMID- 17405775 TI - NICE guideline for Parkinson's disease. PMID- 17405776 TI - Routine thrombolysis with intravenous tissue plasminogen activator in acute ischaemic stroke patients aged 80 years or older: a single centre experience. PMID- 17405777 TI - Why stroke patients don't like Mondays (or Saturdays or Sundays). PMID- 17405778 TI - A cardiovascular benefit of ophthalmic beta-blockade. PMID- 17405779 TI - Activities of 16-membered ring macrolides and telithromycin against different genotypes of erythromycin-susceptible and erythromycin-resistant Streptococcus pyogenes and Streptococcus pneumoniae. AB - OBJECTIVES: To test four 16-membered macrolides (josamycin, spiramycin, midecamycin and rokitamycin) along with other compounds in the same class (erythromycin, clarithromycin, roxithromycin and azithromycin) plus clindamycin and telithromycin, against Streptococcus pyogenes and Streptococcus pneumoniae isolates with well-characterized resistance genotypes. METHODS: Four hundred and eighty-six isolates of S. pyogenes and 375 isolates of S. pneumoniae were assayed for their macrolide susceptibilities and investigated by PCR to detect their different erythromycin resistance genes. All strains had been isolated over the period 2002-2003 from specimens of different human origin obtained in 14 different Italian centres. RESULTS: All 16-membered macrolides showed very low MICs (MIC(50)s and MIC(90)s, < or =0.06-0.5 mg/L) for the erythromycin susceptible isolates and for those with the M phenotype, but the telithromycin MICs for the M-type isolates were at least four times higher (MIC(90)s, 0.5 mg/L). In S. pyogenes, the MIC(50)s of 16-membered macrolides for the cMLS(B) isolates were > or = 256 mg/L, whereas that for telithromycin was 4 mg/L; the MIC(50)s of 16-membered macrolides and telithromycin ranged from < or = 0.06 to 0.5 mg/L for the iMLS(B) isolates with erm(A) and from 0.12 to > or = 256 mg/L for those with erm(B). In S. pneumoniae, the MIC(50)s of the 16-membered macrolides for the cMLS(B) isolates ranged from 0.5 to 128 mg/L, whereas for the iMLS(B) isolates their values ranged from < or = 0.06 to 4 mg/L; the MIC(50)s and MIC(90)s of telithromycin for both the cMLS(B) and the iMLS(B) isolates ranged from < or = 0.06 to 0.12 mg/L. CONCLUSIONS: MICs ranged for all the drugs, except telithromycin, from < or = 0.06 to > or = 256 mg/L, with 15% to 30% resistant S. pyogenes for all drugs tested except clindamycin (8%) and telithromycin (5.4%) and 10% to 40% resistant S. pneumoniae for all drugs tested except telithromycin (0.3%). In both S. pyogenes and S. pneumoniae, erythromycin resistance related to a mef gene meant that telithromycin MICs were definitely higher than in erythromycin-susceptible isolates, although telithromycin susceptibility was preserved in all cases. In S. pyogenes, the activity of both 16-membered macrolides and telithromycin against the iMLS(B) strains proved to be dependent on the erm gene involved, being greater against isolates with erm(A). PMID- 17405780 TI - Evaluation of the in vivo efficacy of topical tobramycin against Pseudomonas sinonasal biofilms. AB - BACKGROUND: There is increasing evidence that bacterial biofilms are present within the sinonasal mucosa of patients with medically recalcitrant chronic rhinosinusitis. The antibiotic concentrations needed to eradicate biofilms are much higher than those commonly used and pose an increased risk for systemic side effects. Topical medications are an alternative approach to deliver high concentrations to the mucosa while limiting systemic side effects. This is the first study to evaluate the effects of a topical antibiotic on Pseudomonas biofilms in an animal model of sinusitis. METHODS: Pseudomonas sinusitis was established in nine rabbits. An irrigation catheter was placed within the lumen of the maxillary sinus, and the rabbits were irrigated with 7 days of normal saline or varying concentrations of tobramycin diluted in normal saline. Bacterial cfu were determined for nasal lavage fluid as well as physically disrupted mucosa, and scanning electron microscopy (SEM) of the mucosal surface was performed at the end of the study. RESULTS: Increasing concentrations of topical tobramycin resulted in the eradication of viable bacteria within the lumen of the sinus but did not eradicate Pseudomonas attached to the mucosa. SEM detected biofilms within the mucosa even in those rabbits treated with high concentrations of topical tobramycin. CONCLUSIONS: Bacterial biofilms attached to the sinonasal mucosa are resistant to topical saline and tobramycin irrigation. SEM was useful in the identification of biofilms, but did not provide a quantitative evaluation for efficacy of treatment. PMID- 17405781 TI - Understanding the differences between contrasting HIV epidemics in east and west Africa: results from a simulation model of the Four Cities Study. AB - OBJECTIVE: To determine if the differences in risk behaviours, the proportions of males circumcised and prevalences of sexually transmitted infections (STIs) observed in two African cities with low prevalence of HIV (Cotonou, Benin, and Yaounde, Cameroon) and two cities with high prevalence (Kisumu, Kenya, and Ndola, Zambia) could explain the contrasting HIV epidemics in the four cities. METHODS: An individual-based stochastic model, STDSIM, was fitted to the demographic, behavioural and epidemiological characteristics of the four urban study populations based on data from the Four Cities Study and other relevant sources. Model parameters pertaining to STI and HIV natural history and transmission were held constant across the four populations. The probabilities of HIV, syphilis and chancroid acquisition were assumed to be doubled among uncircumcised males. A priori plausible ranges for model inputs and outputs were defined and sexual behaviour characteristics, including those pertaining to commercial sex workers (CSWs) and their clients, which were allowed to vary across the sites, were identified based on comparisons of the empirical data from the four sites. The proportions of males circumcised in the model, 100% in Cotonou and Yaounde, 25% in Kisumu and 10% in Ndola, were similar to those observed. A sensitivity analysis was conducted to assess how changes in critical parameters may affect the model fit. RESULTS: Population characteristics observed from the study that were replicated in the model included younger ages at sexual debut and marriage in east Africa compared with west Africa and higher numbers of casual partners in the past 12 months in Yaounde than in the other three sites. The patterns in prevalence of STIs in females in the general population and CSWs were well fitted. HIV prevalence by age and sex and time trends in prevalence in the model were consistent with study data with the highest simulated prevalences in Kisumu and Ndola, intermediate in Yaounde and lowest in Cotonou. The sensitivity analysis suggested that the effect of circumcision on the development of the HIV epidemics may have been mediated indirectly by its effect on ulcerative STI. CONCLUSIONS: The contrasting HIV epidemics in east and west Africa could be replicated in our model by assuming that male circumcision reduced susceptibility to HIV, syphilis and chancroid. Varying rates of male circumcision may have played an important role in explaining the strikingly different HIV epidemics observed in different parts of sub-Saharan Africa. PMID- 17405782 TI - Proportion of new HIV infections attributable to herpes simplex 2 increases over time: simulations of the changing role of sexually transmitted infections in sub Saharan African HIV epidemics. AB - OBJECTIVE: To understand the changing impact of herpes simplex 2 (HSV-2) and other sexually transmitted infections (STIs) on HIV incidence over time in four sub-Saharan African cities, using simulation models. METHODS: An individual-based stochastic model was fitted to demographic, behavioural and epidemiological data from cross-sectional population-based surveys in four African cities (Kisumu, Kenya; Ndola, Zambia; Yaounde, Cameroon; and Cotonou, Benin) in 1997. To estimate the proportion of new HIV infections attributable to HSV-2 and other STIs over time, HIV incidence in the fitted model was compared with that in model scenarios in which the cofactor effect of the STIs on HIV susceptibility and infectivity were removed 5, 10, 15, 20 and 25 years into the simulated HIV epidemics. RESULTS: The proportion of incident HIV attributable to HSV-2 infection (the model estimated population attributable fraction (PAF(M))) increased with maturity of the HIV epidemic. In the different cities, the PAF(M) was 8-31% 5 years into the epidemic, but rose to 35-48% 15 years after the introduction of HIV. In contrast, the proportion of incident HIV attributable to chancroid decreased over time with strongest effects five years after HIV introduction, falling to no effect 15 years after. Sensitivity analyses showed that, in the model, recurrent HSV-2 ulcers had more of an impact on HIV incidence than did primary HSV-2 ulcers, and that the effect of HSV-2 on HIV infectivity may be more important for HIV spread than the effect on HIV susceptibility, assuming that HSV 2 has similar cofactor effects on HIV susceptibility and infectivity. The overall impact of other curable STIs on HIV spread (syphilis, gonorrhoea and chlamydia) remained relatively constant over time. CONCLUSIONS: Although HSV-2 appears to have a limited impact on HIV incidence in the early stages of sub-Saharan African HIV epidemics when the epidemic is concentrated in core groups, it has an increasingly large impact as the epidemic progresses. In generalised HIV epidemics where control programmes for curable STIs are already in place, interventions against HSV-2 may have a key role in HIV prevention. PMID- 17405783 TI - Tubulo-interstitial nephritis with Fanconi syndrome in Behcet disease. PMID- 17405784 TI - Cinacalcet modifies the pH of solutions in vitro: possible implications for gastro-intestinal side effects in vivo. PMID- 17405785 TI - For patients taking herbal therapy--lessons from aristolochic acid nephropathy. PMID- 17405786 TI - Thrombin inhibition with melagatran does not attenuate renal ischaemia reperfusion injury in rats. AB - BACKGROUND: Renal ischaemia-reperfusion (IR) is associated with activation of the coagulation system and inflammation within the kidney. The aim of the present study was to examine the effects of selective thrombin inhibition with melagatran on kidney morphology and function in rats subjected to renal IR. METHODS: Sprague Dawley rats underwent renal IR (35 min of bilateral renal arterial clamping), or sham surgery. Treatment groups were: (i) IR-Saline, (ii) IR-Melagatran, (iii) Sham-Saline, and (iv) Sham-Melagatran. Twenty minutes prior to renal IR, the rats were administered a bolus dose of saline vehicle or melagatran [0.5 mumol/kg, subcutaneously (s.c.)] followed by a continuous infusion throughout (0.08 micromol/kg/h, s.c.). Forty-eight hours after IR, renal function was assessed in anaesthetized animals and kidney histology was analysed semi-quantitatively. RESULTS: Rats in group IR-Saline showed an approximate 85% reduction in glomerular filtration rate, 5-fold increases in fractional urinary excretion rates of sodium, potassium and water, and marked renal histological abnormalities, compared with sham (P < 0.05). Renal histopathological changes in the cortex and outer medulla were characterized by tubular necrosis and atrophy, tubular cast formation and interstitial inflammation. In addition, there was significant vascular congestion in the inner stripe of the outer medullary zone. Melagatran treatment had no significant effects on any of the abnormalities in kidney morphology or function in rats subjected to renal IR. Plasma melagatran concentrations were within a range known to exert significant antithrombotic effects, throughout the study period. CONCLUSIONS: Thrombin inhibition with melagatran did not ameliorate abnormalities in kidney morphology or function 48 h after renal IR. These results indicate that melagatran is not renoprotective in rats subjected to renal IR. PMID- 17405789 TI - Association of low-grade urinary albumin excretion with left ventricular hypertrophy in the general population: a reply. PMID- 17405790 TI - Use of albumin creatinine ratio and urine albumin concentration as a screening test for albuminuria in an Indo-Asian population. AB - BACKGROUND: Albuminuria (>30 mg/day) based on 24 h urine albumin excretion is one of the criteria for chronic kidney disease (CKD) and a predictor of cardiovascular disease (CVD). Differences in urine albumin concentration and creatinine excretion rates between Indo-Asians and other populations may require different threshold values for detection of albuminuria. We compared the use of spot urine albumin concentration and urine albumin to creatinine excretion ratio for detection of albuminuria in this population. METHODS: A total of 577 subjects aged >or=40 years, 54% of whom were women, were recruited from the general population in Karachi, Pakistan. Albumin concentration (mg/l) and albumin to creatinine ratio (mg/g of creatinine) were determined in a spot morning urine sample, and albuminuria (30 mg/day or greater) measured in a 24 h urine collected on the subsequent day. RESULTS: The median (25-75 percentile) of urine albumin excretion was 4.8 (3.6-10.3) mg/day: 5.4 (3.7-12.5) mg/day in men and 4.5 (3.8 8.9) mg/day in women. The overall prevalence (95% CI) of albuminuria was 11.8% (7.2-12.0%): 14.8% in men and 9.2% in women (P = 0.04). The areas under the receiver operator characteristic (ROC) curves for urine albumin concentration were 0.86 (0.82-0.90) and 0.88 (0.84-0.92), respectively, in women and men. The areas under the ROC curves for albumin to creatinine ratio were 0.86 (0.82-0.89) and 0.90 (0.86-0.93), respectively, in women and men. For urine albumin concentration, the sensitivity and specificity were 37 and 97%, respectively, in women and 69 and 94%, respectively, in men at the conventionally recommended value of 2 mg/dl. The discriminator value of urine albumin concentration identified in the analysis was 0.5 mg/dl in women (sensitivity of 87% and specificity of 75%) and 1.7 mg/dl in men (sensitivity of 74% and specificity of 93%). For the albumin to creatinine ratio, the sensitivity and specificity were 46 and 95%, respectively, in women and 60 and 97%, respectively, in men at cut off value of 30 mg/g. CONCLUSION: Both urine albumin concentration and albumin to creatinine ratio are acceptable tests for population screening for albuminuria in Indo-Asians. While sensitivities may be suboptimal, particularly in women, lowering the existing thresholds would compromise specificity. Those who screen positive need evaluation and management of CKD and prevention of CVD. PMID- 17405791 TI - Reverse mid-dilution: new way to remove small and middle molecules as well as phosphate with high intrafilter convective clearance. AB - BACKGROUND: The removal of small and middle molecules has a relevant impact on haemodialysis (HD) patient survival. Mid-dilution (MD) is a technique combining ease of use with high diffusive-convective clearances. However, MD may increase the intrafilter blood pressure due to the high filtration fraction. We devised a new filter configuration, reverse MD, with an inverted blood inlet and outlet. We compared biochemical and technical performances of reverse MD vs standard MD. METHODS: Eight HD patients underwent one standard MD treatment and one reverse MD. Samples for instantaneous clearance and total mass removed from dialysate spilling (urea, phosphate, beta2-microglobulin, angiogenin) were obtained. Dialysate and blood pressures in the circuit were monitored every 15 min. The reinfusion rate was set at 6 l/h for both treatments. RESULTS: Absolute removals were very high and statistically comparable in both the configurations. Pressures were significantly lower with the reverse compared with the standard MD: inlet blood pressure was 731+/-222 and 595+/-119 mmHg in the standard and in the reverse MD, respectively. The transmembrane pressures were lower in the reverse compared with the standard MD (422+/-90 and 611+/-136 mmHg for 1st stage; 188+/ 54 and 307+/-56 mmHg for 2nd stage). CONCLUSIONS: Reverse MD could be an ideal technique for high ultrafiltration routine treatments without any external fluid reinfusion. It allows a very high removal of small and middle molecules, with relatively lower intrafilter pressures. PMID- 17405792 TI - Mycophenolate mofetil in induction and maintenance therapy of severe lupus nephritis: a meta-analysis of randomized controlled trials. AB - BACKGROUND: The outcomes of previous trials of mycophenolate mofetil (MMF) in treating severe lupus nephritis (LN) are not in exact agreement. This meta analysis of randomized controlled trials (RCTs) assesses the benefits and harms of MMF in the induction and maintenance therapy of severe LN. METHODS: We searched Medline, EMBASE and the Cochrane Collaboration Database for RCTs that compared MMF with other immunorepressive regimens for treating lupus nephritis and extracted data for remissions, side effects and prognosis in induction therapy and prognosis and side effects in maintenance therapy, and we summarized the combined results of the data of the RCTs as relative risk (RR). RESULTS: We analysed five RCTs with 307 patients-four RCTS providing the data for comparing MMF with cyclophosphamide (CYC) for induction therapy and two RCTs providing the data for comparing MMF with azathioprine (AZA) for maintenance therapy of severe LN. Overall, compared with CYC, induction therapy with MMF reduced the risk of infection significantly (RR 0.65, P<0.001). It also significantly increased the complete remission rate compared with intravenous CYC (RR 3.10, P=0.006). Compared with intravenous CYC, induction therapy with MMF reduced the incidence of leucopenia significantly (RR 0.66, P=0.04). The prognosis and other side effects were not significantly different between MMF and CYC induction therapies. There was no significant difference between the patients receiving MMF and those receiving AZA for maintenance therapy in prognosis or the risks of amenorrhoea and herpes zoster. CONCLUSIONS: MMF has higher efficacy in inducing remission in severe LN than pulsed intravenous therapy with CYC. Induction therapy with MMF is also associated with fewer side effects than induction therapy with CYC. Compared with AZA, MMF also is an alternative for maintenance therapy of severe LN without significant difference in the prognosis or risks of amenorrhoea and herpes zoster. PMID- 17405793 TI - Lanthanum carbonate, body lanthanum accumulation and potential liver toxicity. PMID- 17405794 TI - Use of pure bicarbonate-buffered peritoneal dialysis fluid reduces the incidence of CAPD peritonitis. AB - BACKGROUND: Advances in bag connection technology have reduced the incidence of peritonitis in CAPD patients but there is little information on the effect of the new peritoneal dialysis fluids. METHODS: We studied the incidence of CAPD peritonitis for about 3 years in 100 incident patients--50 patients dialysed with lactate-buffered solution, pH 5.5 and containing glucose degradation products (GDP) (lactate group), and 50 patients with pure bicarbonate-buffered solution, pH 7.4 and low GDP (bicarbonate group). Patients in both groups were similar in age, sex, length of time on CAPD, connection technology and handling of dialysis. RESULTS: In the lactate group, 74 episodes of peritonitis were recorded compared with 43 in the bicarbonate group, i.e. one episode per 21 patient-months with the lactate dialysis fluid and one episode per 36 patient-months with the bicarbonate dialysis fluid (OR 0.58, 95% CI 0.37-0.91, P = 0.017). A total of 3369 exchanges per episode of peritonitis were recorded for bicarbonate compared with 2004 exchanges per episode of peritonitis in the lactate group. The majority of organisms isolated in both groups were Gram-positive bacteria, with a predominance of the oropharyngeal and cutaneous endogenous flora. Three episodes of fungal peritonitis occurred in the lactate group and none in the bicarbonate group. CONCLUSIONS: Our results suggest that the pure bicarbonate-buffered peritoneal dialysis fluid appears to reduce the frequency of peritonitis in CAPD patients possibly in relation to greater biocompatibility and maintenance of peritoneal membrane structural integrity. Similar results can probably relate to all low-GDP solutions. PMID- 17405795 TI - Role of histone modification in chromatin dynamics. AB - The covalent modification of histone tails has emerged as a crucial step in controlling the eucaryotic genomes. Eucaryotic cells must possess mechanisms for condensing and decondensing chromatin. Moreover, chromatin condensation is particularly evident during mitosis and apoptotic cell death, whereas chromatin relaxation is necessary for replication, repair, recombination and transcription. The post-translational modifications of histone tails such as reversible acetylation, phosphorylation and methylation play a critical role in dynamic condensation/relaxation that occurs during the cell cycle. Histone phosphorylation is believed to play a direct role in mitosis, cell death, repair, replication and recombination. In this review, we discuss recent progress in studies of histone phosphorylation. PMID- 17405796 TI - Phosphorylation of the C-terminal domain of RNA polymerase II plays central roles in the integrated events of eucaryotic gene expression. AB - RNA polymerase II (Pol II) is the only polymerase to possess heptapeptide repeats in the C-terminal domain (CTD) of its large subunit. During transcription, CTD phopshorylation occurs and is maintained from initiation to termination. To date, among the three known CTD kinases possessing CDK-cyclin pairs, TFIIH is the only one that forms a preinitiation complex. The Mediator complex plays essential roles in transcription initiation and during the transition from initiation to elongation by transmitting signals from transcriptional activators to Pol II. P TEFb specifically plays a role in transcription elongation. TFIIH and mediator phosphorylate serine 5 (Ser5) of the CTD heptapeptide repeat sequence, whereas P TEFb phosphorylates serine 2 (Ser2). Recently, it has become clear that CTD phosphorylation is not only essential for transcription, but also as a platform for RNA processing and chromatin regulation. In this review, we discuss the central role of Pol II phosphorylation in these nuclear events. PMID- 17405797 TI - Molecular mechanism of the inhibitory effect of cobalt ion on thermolysin activity and the suppressive effect of calcium ion on the cobalt ion-dependent inactivation of thermolysin. AB - Thermolysin activity in the hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-l leucine amide (FAGLA) and FA-l-leucyl-l-alanine amide (FALAA) was examined at various Co(2+) and Ca(2+) concentrations. It decreased to 28% with increasing [Co(2+)] up to 18 mM. The Co(2+)-dependent inactivation was in part suppressed by adding Ca(2+) ion up to 0.5 mM, but 33% of the activity remained to be inactivated even with a sufficient concentration of Ca(2+) (>0.5 mM). The Co(2+) dependent inactivation was shown to be composed of Ca(2+)-sensitive and Ca(2+) insensitive parts. In the latter part which is observed at [Ca(2+)] >0.5 mM, Co(2+) plays as a competitive inhibitor. On the other hand, the Co(2+)-dependent inactivation in the Ca(2+)-sensitive part observed at [Ca(2+)] <0.5 mM proceeds time-dependently following second-order kinetics, and the time-course is in good agreement with that of decrease in the thermolysin band due to autolysis in SDS PAGE. This indicates that Co(2+) accelerates the autolysis. Here, we describe the co-regulation of thermolysin activity by Co(2+) and Ca(2+) ions and propose a molecular mechanism for the inhibition of thermolysin by Co(2+) and suppressive effect of Ca(2+) on the Co(2+)-dependent inhibition. Co(2+) ion inhibits thermolysin activity not only as a competitive inhibitor but also promoting the autolysis. PMID- 17405798 TI - Kinetic analysis of the activation-and-inhibition dual effects of cobalt ion on thermolysin activity. AB - Thermolysin activity as well as its stability is remarkably enhanced by high concentration of neutral salts consisting of Na(+), K(+), Cl(-) and Br(-) in the synthesis and hydrolysis of N-carbobenzoxy-L-aspertyl-L-phenylalanine methyl ester and hydrolysis of N-[3-(2-furyl)acryloyl]-glycyl-L-leucine amide (FAGLA) [Inouye, K. (1992) J. Biochem. 112, 335-340]. However, effect of divalent salts on thermolysin activity has not been investigated systematically. In this study, effect of Co(2+) ion on thermolysin activity in the hydrolysis of FAGLA was examined. Thermolysin activity increased 3-4 times with increasing the Co(2+) concentration to 2 mM, but the enhanced activity was considerably reduced with higher Co(2+) concentration (2-18 mM). The activation-and-inhibition dual effects of Co(2+) ion were analysed kinetically. Release of the catalytic Zn(2+) ion from thermolysin, concomitantly occurred with the Co(2+)-dependent activation, was measured with a Zn(2+)-specific fluorescent probe. This indicates that the activation is caused by substituting Co(2+) ion for the catalytic Zn(2+) ion. Meanwhile, the Co(2+)-dependent activation was inhibited competitively by Zn(2+) ion (0.1-1.0 muM) added, similarly to that it is inhibited by higher concentration of Co(2+) ion. These lines of evidence provide a strategy for regulating thermolysin activity with Co(2+) and Zn(2+) ions. PMID- 17405799 TI - Effects of site-directed mutagenesis of the surface residues Gln128 and Gln225 of thermolysin on its catalytic activity. AB - Thermolysin is remarkably activated and stabilized by neutral salts with varying degrees depending on salt species, and particular surface residues are thought to be especially important in its activity and stability [Inouye, K. (1992) J. Biochem. 112, 335-340; Inouye, K. et al. (1998) Biochim. Biophys. Acta 1388, 209 214]. In this study, we examined the mutational effects of the surface residues of thermolysin. Gln128 and Gln225 were selected as the residues to be mutated because they are located on the surface loop and close to but not in the active site (23.5 and 15.8 A far from the active site zinc ion, respectively) and fully solvent accessible. Nine single mutants [Q128K (Gln128 is replaced with Lys), Q128E, Q128A, Q225K, Q225R, Q225E, Q225D, Q225A and Q225V] were constructed by site-directed mutagenesis. Mutational changes in catalytic activity were found only in the mutant thermolysins having a hydrophobic residue at the position 225 (Q225A and Q225V). In the hydrolysis of a neutral substrate N-[3-(2 furyl)acryloyl]-glycyl-l-leucine amide (FAGLA), the alkaline pK(a) value of Q225A is 8.48 +/- 0.04, being higher by 0.42 +/- 0.07 units than that of the wild-type thermolysin. The k(cat)/K(m) value of the wild-type enzyme is enhanced 14 times with 4 M NaCl, and those of Q225A and Q225V are enhanced 10 and 19 times, respectively. In the hydrolysis of a negatively charged substrate N-carbobenzoxy l-aspartyl-l-phenylalanine methyl ester (ZDFM), unlike FAGLA, the initial velocities of Q225A and Q225V decreased to 30 and 50% of that of the wild-type enzyme, respectively. Their thermal stability is similar to that of the wild-type enzyme. These findings indicate that even a single mutation at the thermolysin surface induces changes in the electrostatic environment in the active site and affects the activity. Thus, site-directed mutagenesis of surface residues of thermolysin, including apparently thermodynamically unfavorable introduction of hydrophobic residues, should be explored to improve its activity and stability. PMID- 17405800 TI - Efficacy and safety of multiple intravitreal triamcinolone injections for refractory diabetic macular oedema. AB - AIM: The efficacy and safety of repeated injections of intravitreal triamcinolone (IVTA) for diabetic macular oedema is unclear, with results of previous reports conflicting. METHODS: This is a prospective, observational case series of 27 eyes receiving IVTA for diabetic macular oedema. LogMAR visual acuity (VA) and central macular thickness (CMT) were measured at baseline and in 3 to 6 monthly intervals for up to 24 months, then correlated with the number of IVTA injections given. RESULTS: One IVTA injection was required in 6 (18%) eyes, 2 in 8 (24%) eyes, 3 in 13 (39%) eyes and 4-5 in 6 (18%) eyes. VA improved in all patients, but neither the final improvement in VA nor the absolute improvement in CMT from baseline to 24 months correlated with the number of injections received (p = 0.44 and 0.84, respectively). Cataract surgery was more frequent in eyes receiving more injections (p = 0.01). CONCLUSIONS: This study suggests that repeated injections of IVTA continue to be as effective as the first over a 2-year period. The probability of cataract surgery increases with an increasing number of injections. PMID- 17405801 TI - Optical coherence tomographic artefacts in diseases of the retinal pigment epithelium. AB - AIMS: To show optical coherence tomography (OCT) artefacts in images from patients with retinal pigment epithelium detachment and retinal laser scars when OCT protocol analyses were applied. METHODS: All OCT retinal scans using OCT-3000 (software 4.02) were reviewed over a three-month period. 13 eyes of 11 patients were selected for this study. 10 eyes had retinal pigment epithelial detachments and 3 had retinal laser scars. All patients had ophthalmic examination, fluorescein angiography (one had indocyanine green angiography) and OCT. All OCT processing and analysis protocols were applied in each case. RESULTS: 10 eyes of 8 patients with retinal pigment epithelial detachments showed flattening of the retinal pigment epithelium and apparent inversion of the dome of the detachment when scan protocol analyses were applied. 3 eyes with retinal laser scars displayed thinning of the retinal pigment epithelium without changes behind the scar. The retinal tissues around the lesions did not show any alteration. CONCLUSIONS: OCT scan analysis is an excellent method to obtain specific information about the retina. However, some lesions that cause disruption of external reflectivity (retinal pigment epithelium) can cause software-related artefacts when analysis protocols are applied. To prevent diagnostic error, re evaluation of the clinical fundus examination should be considered in any patient in whom OCT findings do not appear consistent with the initial clinical findings. PMID- 17405802 TI - Excess Indigenous mortality: are Indigenous Australians more severely disadvantaged than other Indigenous populations? AB - BACKGROUND: International targets for reducing health inequalities, such as the Millennium Development Goals, are stated in terms of national targets. However, dramatic health differentials exist within countries, even developed ones. Studies indicate that the Indigenous population of Australia suffers a life expectancy disadvantage greater than differentials found in Indigenous populations of other developed countries. We re-examine recent national mortality levels and trends of Indigenous Australians. METHODS: Analyses of Indigenous mortality are plagued by 'numerator-denominator bias', whereby reporting of Indigenous status differs in deaths (numerators) and population (denominators). We apply demographic evaluation methods developed to address such problems to data from the 1991, 1996 and 2001 censuses of Australia and to the death registration data for the period. RESULTS: The propensity of Australia's population to report Indigenous status increased between each census, particularly between 1991 and 1996, while recording of deaths as Indigenous increased sharply. Adjusted for bias, the Indigenous population had a life expectancy approximately 13 years below that of the non-Indigenous population, a 2-year greater disadvantage than recently estimated for the Maori in New Zealand. Indigenous mortality fell during the 1990s, but slightly more slowly than that of non-Indigenous Australians, leaving differentials slightly increased. CONCLUSIONS: Around the world Indigenous populations are estimated to suffer a mortality disadvantage compared with non-Indigenous populations. However, establishing the magnitude of and trend in the disadvantage is difficult because of bias. Using appropriate methods to adjust for bias, the Indigenous population of Australia is estimated to suffer a life expectancy shortfall of about 13 years, greater than similar gaps in other developed countries. PMID- 17405803 TI - Sensory-specific balance training in older adults: effect on position, movement, and velocity sense at the ankle. AB - BACKGROUND AND PURPOSE: Age-related changes in proprioception contribute to impairments in postural control and increased fall risk in older adults. The purpose of this randomized controlled trial was to examine the effects of balance exercises on proprioception. SUBJECTS: The participants were 36 older people and 24 younger people who were healthy. METHODS: Older participants were randomly assigned to a balance exercise group (n=17) or a falls prevention education group (n=19). Baseline, postintervention, and 8-week follow-up measurements of 3 proprioceptive measures (threshold to perception of passive movement, passive joint position sense, and velocity discrimination) were obtained at the ankle. For comparative purposes, younger participants underwent a one-time assessment of the 3 proprioceptive measures. RESULTS: Postintervention improvements in velocity discrimination were found in the balance exercise group when compared with values at baseline and in the falls prevention education group. Age-related differences found at baseline were reduced in the balance exercise group after intervention. Improvements were not maintained at the 8-week follow-up. Threshold to perception of passive movement and passive joint position sense did not change as a function of the exercise intervention. DISCUSSION AND CONCLUSION: The results suggest that short-term improvements in velocity sense, but not movement and position sense, may be achieved following a balance exercise intervention. PMID- 17405804 TI - Biomechanics of submaximal recumbent cycling in adolescents with and without cerebral palsy. AB - BACKGROUND AND PURPOSE: The purpose of this study was to compare the biomechanics of recumbent cycling between adolescents with cerebral palsy (CP) classified at Gross Motor Function Classification System (GMFCS) levels III and IV and adolescents with typical development (TD). SUBJECTS: Twenty subjects, ages (X+/ SD) 15.2+/-1.6 years (10 with TD, 10 with CP), participated. METHODS: Lower extremity kinematics and muscle activity were measured at 30 and 60 rpm while subjects pedaled on a recumbent cycle. Energy expenditure and perceived exertion were measured during a 5-minute test, and efficiency was calculated. Noncircular data were analyzed with analyses of variance. Circular data were analyzed using circular t tests. RESULTS: Differences were found between groups for joint kinematics for all motions. Subjects with CP displayed earlier onsets and later offsets of muscle activity, increased co-contraction of agonist and antagonist muscles, and decreased efficiency compared with subjects with TD. There were no differences in perceived exertion. DISCUSSION AND CONCLUSION: Differences in cycling biomechanics between children with CP and children with TD may be due to decreased strength and motor control in the children with CP. PMID- 17405805 TI - Evaluation of two preventive interventions for reducing musculoskeletal complaints in operators of video display terminals. AB - BACKGROUND AND PURPOSE: The purpose of this study was to evaluate the efficacy of a preventive ergonomic intervention, which was provided by physical therapists, on spinal and upper-extremity work-related posture and symptom complaints of workers who use video display terminals (VDT). SUBJECTS: Two hundred employees who spent at least 20 hours per week at a VDT were randomly divided into 2 groups. Group E received the ergonomic intervention and an informative brochure, and group I received only the brochure. METHODS: Both groups were evaluated at the beginning of the study and at a follow-up 5 months later. The following tools were used: a pain drawing and the Rapid Entire Body Assessment (REBA) method to assess spinal and upper-extremity work-related posture. RESULTS: Group E had a lower REBA score and reduced lower back, neck, and shoulder symptoms compared with group I. DISCUSSION AND CONCLUSION: The results suggest that a personalized preventive ergonomic intervention can improve spinal and upper-extremity work related posture and musculoskeletal symptoms for workers who use VDTs. PMID- 17405806 TI - Effect of aerobic training on walking capacity and maximal exercise tolerance in patients with multiple sclerosis: a randomized crossover controlled study. AB - BACKGROUND AND PURPOSE: Physical deconditioning is involved in the impaired exercise tolerance of patients with multiple sclerosis (MS), but data on the effects of aerobic training (AT) in this population are scanty. The purpose of this study was to compare the effects of an 8-week AT program on exercise capacity-in terms of walking capacity and maximum exercise tolerance, as well as its effects on fatigue and health-related quality of life-as compared with neurological rehabilitation (NR) in subjects with MS. SUBJECTS AND METHODS: Nineteen subjects (14 female, 5 male; mean age [X+/-SD]=41+/-8 years) with mild to moderate disability secondary to MS participated in a randomized crossover controlled study. Eleven subjects (8 female, 3 male; mean age [X+/-SD]=44+/-6 years) completed the study. RESULTS: After AT, but not NR, the subjects' walking distances and speeds during a self-paced walk were significantly improved, as were their maximum work rate, peak oxygen uptake, and oxygen pulse during cardiopulmonary exercise tests. The increases in peak oxygen uptake and maximum work rate, but not in walking capacity, were significantly higher after AT, as compared with after NR. Additionally, the subjects who were most disabled tended to benefit more from AT. There were no differences between AT and NR in effects on fatigue, and the results showed that AT may have partially affected health related quality of life. DISCUSSION AND CONCLUSION: The results suggest that AT is more effective than NR in improving maximum exercise tolerance and walking capacity in people with mild to moderate disability secondary to MS. PMID- 17405807 TI - Communicating with patients: what happens in practice? AB - BACKGROUND AND PURPOSE: Communication is the most important aspect of practice that health care professionals have to master. The purpose of this study was to measure the content and prevalence of verbal and nonverbal communications between physical therapists and patients with back pain. SUBJECTS: Seven physical therapists and 21 patients with back pain participated in this study. METHODS: The first interaction following the initial assessment was recorded with a video camera. The outcome measures were the Medical Communications Behavior System (verbal communication) and frequencies of nonverbal behaviors (affirmative head nodding, smiling, eye gaze, forward leaning, and touch). Semistructured interviews were undertaken with the physical therapists to determine the perceived influence of the video camera. RESULTS: A total of 2,055 verbal statements were made. Physical therapists spent approximately twice as much time talking as patients, with content behaviors (such as taking history and giving advice) comprising 52% of verbal communications. The most prevalent nonverbal behaviors were touch by physical therapists (54%) and eye gaze by patients (84%). DISCUSSION AND CONCLUSION: The prevalence and content of communication can be measured with video analysis and validated tools. Communication is an extremely important but underexplored dimension of the patient-therapist relationship, and the methods described here could provide a useful model for further research and reflective practice. PMID- 17405808 TI - Tai chi and perceived health status in older adults who are transitionally frail: a randomized controlled trial. AB - BACKGROUND AND PURPOSE: Tai chi, a Chinese exercise derived from martial arts, while gaining popularity as an intervention for reducing falls in older adults, also may improve health status. The purpose of this study was to determine whether intense tai chi (TC) exercise could improve perceived health status and self-rated health (SRH) more than wellness education (WE) for older adults who are transitionally frail. SUBJECTS: Study subjects were 269 women who were >or=70 years of age and who were recruited from 20 congregate independent senior living facilities. METHODS: Participants took part in a 48-week, single-blind, randomized controlled trial. They were randomly assigned to receive either TC or WE interventions. Participants were interviewed before randomization and at 1 year regarding their perceived health status and SRH. Perceived health status was measured with the Sickness Impact Profile (SIP). RESULTS: Compared with WE participants, TC participants reported significant improvements in the physical dimension and ambulation categories and borderline significant improvements in the body care and movement category of the SIP. Self-rated health did not change for either group. DISCUSSION AND CONCLUSION: These findings suggest that older women who are transitionally frail and participate in intensive TC exercise demonstrate perceived health status benefits, most notably in ambulation. PMID- 17405809 TI - Sustained cell polarity and virulence in the phytopathogenic fungus Ustilago maydis depends on an essential cyclin-dependent kinase from the Cdk5/Pho85 family. AB - Cyclin-dependent kinases from the Cdk5/Pho85 family are thought to play important roles in morphogenesis in organisms as diverse as yeast and humans. Here we used the corn smut fungus Ustilago maydis to address the role of Cdk5/Pho85 kinases in the morphogenesis and virulence of dimorphic phytopathogens. We found that Cdk5 is essential for growth in U. maydis. A temperature-sensitive cdk5 mutant caused cell wall and morphology defects at the restrictive temperature. Actin patches labeled with a fimbrin-GFP fusion protein were delocalized and a GFP-Myo5 fusion was directed towards the growing cell pole and rapidly dissociated from the tip. These defects were found to be due to an impairment in the maintenance of cell polarity. Our results indicated that Cdk5 is required for the activity of Rac1, probably at the level of the localization of its GEF, Cdc24. Cdk5 was required for full virulence, probably because mutant cells are unable to sustain the dramatic polar growth required for the formation of the infective structures. These results support a major role for morphogenesis in the virulence program of dimorphic fungi. PMID- 17405810 TI - Stuck in reverse: loss of LC1 in Trypanosoma brucei disrupts outer dynein arms and leads to reverse flagellar beat and backward movement. AB - Axonemal dyneins are multisubunit molecular motors that provide the driving force for flagellar motility. Dynein light chain 1 (LC1) has been well studied in Chlamydomonas reinhardtii and is unique among all dynein components as the only protein known to bind directly to the catalytic motor domain of the dynein heavy chain. However, the role of LC1 in dynein assembly and/or function is unknown because no mutants have previously been available. We identified an LC1 homologue (TbLC1) in Trypanosoma brucei and have investigated its role in trypanosome flagellar motility using epitope tagging and RNAi studies. TbLC1 is localized along the length of the flagellum and partitions between the axoneme and soluble fractions following detergent and salt extraction. RNAi silencing of TbLC1 gene expression results in the complete loss of the dominant tip-to-base beat that is a hallmark of trypanosome flagellar motility and the concomitant emergence of a sustained reverse beat that propagates base-to-tip and drives cell movement in reverse. Ultrastructure analysis revealed that outer arm dyneins are disrupted in TbLC1 mutants. Therefore LC1 is required for stable dynein assembly and forward motility in T. brucei. Our work provides the first functional analysis of LC1 in any organism. Together with the recent findings in T. brucei DNAI1 mutants [Branche et al. (2006). Conserved and specific functions of axoneme components in trypanosome motility. J. Cell Sci. 119, 3443-3455], our data indicate functionally specialized roles for outer arm dyneins in T. brucei and C. reinhardtii. Understanding these differences will provide a more robust description of the fundamental mechanisms underlying flagellar motility and will aid efforts to exploit the trypanosome flagellum as a drug target. PMID- 17405811 TI - The control of tissue architecture over nuclear organization is crucial for epithelial cell fate. AB - The remodeling of nuclear organization during differentiation and the dramatic alteration of nuclear organization associated with cancer development are well documented. However, the importance of tissue architecture in the control of nuclear organization remains to be determined. Differentiation of mammary epithelial cells into functional tissue structures, in three-dimensional culture, is characterized by a specific tissue architecture (i.e. a basoapical polarity axis), cell cycle exit and maintenance of cell survival. Here we show that induction of partial differentiation (i.e. basal polarity only, cell cycle exit and cell survival) by epigenetic mechanisms in malignant breast cells is sufficient to restore features of differentiation-specific nuclear organization, including perinucleolar heterochromatin, large splicing factor speckles, and distinct nuclear mitotic apparatus protein (NuMA) foci. Upon alteration of nuclear organization using an antibody against NuMA, differentiated non neoplastic cells undergo apoptosis, whereas partially differentiated malignant cells enter the cell cycle. Non-neoplastic cells cultured under conditions that prevent the establishment of apical polarity also enter the cell cycle upon NuMA antibody treatment. These findings demonstrate that the differentiation status rather than the non-neoplastic or neoplastic origin of cells controls nuclear organization and suggest a link between nuclear organization and epigenetic mechanisms dictated by tissue architecture for the control of cell behavior. PMID- 17405812 TI - Dose-dependent inhibition of proteasome activity by a mutant ubiquitin associated with neurodegenerative disease. AB - The ubiquitin-proteasome system is the main regulated intracellular proteolytic pathway. Increasing evidence implicates impairment of this system in the pathogenesis of diseases with ubiquitin-positive pathology. A mutant ubiquitin, UBB(+1), accumulates in the pathological hallmarks of tauopathies, including Alzheimer's disease, polyglutamine diseases, liver disease and muscle disease and serves as an endogenous reporter for proteasomal dysfunction in these diseases. UBB(+1) is a substrate for proteasomal degradation, however it can also inhibit the proteasome. Here, we show that UBB(+1) properties shift from substrate to inhibitor in a dose-dependent manner in cell culture using an inducible UBB(+1) expression system. At low expression levels, UBB(+1) was efficiently degraded by the proteasome. At high levels, the proteasome failed to degrade UBB(+1), causing its accumulation, which subsequently induced a reversible functional impairment of the ubiquitin-proteasome system. Also in brain slice cultures, UBB(+1) accumulation and concomitant proteasome inhibition was only induced at high expression levels. Our findings show that by varying UBB(+1) expression levels, the dual proteasome substrate and inhibitory properties can be optimally used to serve as a research tool to study the ubiquitin-proteasome system and to further elucidate the role of aberrations of this pathway in disease. PMID- 17405813 TI - RhoB plays an essential role in CXCR2 sorting decisions. AB - The CXCR2 chemokine receptor is a G-protein-coupled receptor that undergoes clathrin-mediated endocytosis upon ligand binding. The trafficking of CXCR2 is crucial for cells to maintain a proper chemotactic response. The mechanisms that regulate the recycling/degradation sorting decision are unknown. In this study, we used dominant-negative (T19N) and GTPase-deficient activated (Q63L) RhoB mutants, as well as RhoB small interfering RNA (siRNA) to investigate the role of RhoB in CXCR2 trafficking. Expression of either of the RhoB mutants or transfection of RhoB siRNA impaired CXCR2-mediated chemotaxis. Expression of RhoB T19N and transfection of RhoB siRNA impaired sorting of CXCR2 to the lysosome after 3 hours of CXCL8 stimulation and impaired CXCL8-induced CXCR2 degradation. In cells expressing the RhoB Q63L mutant, CXCR2 recycling through the Rab11a recycling compartment was impaired after 30 minutes of CXCL8 stimulation as was CXCL8-induced CXCR2 degradation. For cells expressing activated RhoB, CXCR2 colocalized with Rab4, a marker for the rapid recycling pathway, and with the mannose-6-phosphate receptor, which traffics between the trans-Golgi network and endosomes. These data suggest that CXCR2 recycles through alternative pathways. We conclude that oscillation of RhoB GTPase activity is essential for appropriate sorting decisions, and for directing CXCR2 degradation and recycling--events that are required for optimal chemotaxis. PMID- 17405814 TI - Symmetrical localization of extrachromosomally replicating viral genomes on sister chromatids. AB - In eukaryotes, many latent viruses replicate as extrachromosomal molecules, called episomes, and efficiently segregate to daughter cells by noncovalently attaching to mitotic chromosomes. To understand the mechanism governing the processes, we analyzed the detailed subcellular localization of Epstein-Barr virus (EBV) genomes and a viral protein EBNA1, a bridging molecule between viral genomes and cellular chromatin. In the cells that were infected with a recombinant EBV expressing epitope-tagged EBNA1, EBNA1 localized to intranuclear punctate dots, which coincided with the localization of EBV genomes as revealed by fluorescence in situ hybridization (FISH). A significant number of EBNA1 dots were found to localize symmetrically on sister chromatids of mitotic chromosomes. Such symmetrical localization of EBNA1 dots was observed in prematurely condensed G2 chromosomes as well, correlating with the presence of closely spaced double dots of EBNA1 in G2-phase-enriched cells. The EBNA1 double dots were occasionally interconnected by the FISH signals of EBV episomes, exhibiting a dumbbell-like appearance. Thus, we propose that the partitioning of EBNA1 molecules onto sister chromatids during cellular DNA replication underlies the non-stochastic segregation of extrachromosomally replicating viral genomes. PMID- 17405815 TI - Inflammatory ROS promote and cooperate with the Fanconi anemia mutation for hematopoietic senescence. AB - The proinflammatory cytokine tumor necrosis factor alpha (TNFalpha) inhibits hematopoietic stem cell (HSC) expansion, interferes with HSC self-renewal and compromises the ability of HSC to reconstitute hematopoiesis. We have investigated mechanisms by which TNFalpha suppresses hematopoiesis using the genomic instability syndrome Fanconi anemia mouse model deficient for the complementation-group-C gene (Fancc). Examination of senescence makers, such as senescence-associated beta-galactosidase, HP1-gamma, p53 and p16(INK4A) shows that TNFalpha induces premature senescence in bone marrow HSCs and progenitor cells as well as other tissues of Fancc-/- mice. TNFalpha-induced senescence correlates with the accumulation of reactive oxygen species (ROS) and oxidative DNA damage. Neutralization of TNFalpha or deletion of the TNF receptor in Fancc-/ mice (Fancc-/-;Tnfr1-/-) prevents excessive ROS production and hematopoietic senescence. Pretreatment of TNFalpha-injected Fancc-/- mice with a ROS scavenger significantly reduces oxidative base damage, DNA strand breaks and senescence. Furthermore, HSCs and progenitor cells from TNFalpha-treated Fancc-/- mice show increased chromosomal aberrations and have an impaired oxidative DNA-damage repair. These results indicate an intimate link between inflammatory reactive oxygen species and DNA-damage-induced premature senescence in HSCs and progenitor cells, which may play an important role in aging and anemia. PMID- 17405816 TI - A regulatory role for CRM1 in the multi-directional trafficking of splicing snRNPs in the mammalian nucleus. AB - Distinct pathways of ribonucleoprotein transport exist within the nucleus, connected to their biogenesis and maturation. These occur despite evidence that the major mechanism for their movement within the nucleus is passive diffusion. Using fusions of Sm proteins to YFP, CFP and photoactivatable GFP, I have demonstrated that pathways with uni-directional bulk flow of complexes can be maintained within the nucleus despite multi-directional exchange of individual complexes. Newly imported splicing small nuclear ribonucleoproteins (snRNPs) exchange between Cajal bodies (CBs) within a nucleus and access the cytoplasm, but are unable to accumulate in speckles. By contrast, snRNPs at steady-state exchange freely in any direction between CBs and speckles, but cannot leave the nucleus. In addition to these surprising qualitative observations in the behaviour of nuclear complexes, sensitive live-cell microscopy techniques can detect subtle quantitative disturbances in nuclear dynamics before they have had an effect on overall nuclear organization. Inhibition of the nuclear export factor, CRM1, using leptomycin B results in a change in the dynamics of interaction of newly imported snRNPs with CBs. Together with the detection of interactions of CRM1 with Sm proteins and the survival of motor neurons (SMN) protein, these studies suggest that the export receptor CRM1 is a key player in the molecular mechanism for maintaining these pathways. Its role in snRNP trafficking, however, appears to be distinct from its previously identified role in small nucleolar RNP (snoRNP) maturation. PMID- 17405817 TI - Functional importance of polymerization and localization of calsequestrin in C. elegans. AB - Dual roles of calsequestrin (CSQ-1) being the Ca2+ donor and Ca2+ acceptor make it an excellent Ca2+-buffering protein within the sarcoplasmic reticulum (SR). We have isolated and characterized a calsequestrin (csq-1)-null mutant in Caenorhabditis elegans. To our surprise, this mutant csq-1(jh109) showed no gross defects in muscle development or function but, however, is highly sensitive to perturbation of Ca2+ homeostasis. By taking advantage of the viable null mutant, we investigated the domains of CSQ-1 that are important for polymerization and cellular localization, and required for its correct buffering functions. In transgenic animals rescued with various CSQ-1 constructs, the in vivo patterns of polymerization and localization of several mutated calsequestrins were observed to correlate with the structure-function relationship. Our results suggest that polymerization of CSQ-1 is essential but not sufficient for correct cellular localization and function of CSQ-1. In addition, direct interaction between CSQ-1 and the ryanodine receptor (RyR) was found for the first time, suggesting that the cellular localization of CSQ-1 in C. elegans is indeed modulated by RyR through a physical interaction. PMID- 17405818 TI - Crosstalk between neovessels and mural cells directs the site-specific expression of MT1-MMP to endothelial tip cells. AB - The membrane-anchored matrix metalloproteinase MT1-MMP (also known as Mmp14) plays a key role in the angiogenic process, but the mechanisms underlying its spatiotemporal regulation in the in vivo setting have not been defined. Using whole-mount immunohistochemical analysis and the lacZ gene inserted into the Mmp14 gene, we demonstrate that MT1-MMP vascular expression in vivo is confined largely to the sprouting tip of neocapillary structures where endothelial cell proliferation and collagen degradation are coordinately localized. During angiogenesis in vitro, wherein endothelial cells are stimulated to undergo neovessel formation in the presence or absence of accessory mural cells, site specific MT1-MMP expression is shown to be controlled by crosstalk between endothelial cells and vascular smooth muscle cells (VSMC). When vessel maturation induced by VSMCs is inhibited by introducing a soluble form of the receptor tyrosine kinase Tek, MT1-MMP distribution is no longer restricted to the endothelial tip cells, but instead distributes throughout the neovessel network in vitro as well as ex vivo. Taken together, these data demonstrate that vascular maturation coordinated by endothelial cell/mural cell interactions redirects MT1 MMP expression to the neovessel tip where the protease regulates matrix remodeling at the leading edge of the developing vasculature. PMID- 17405819 TI - Digital ischemic changes after gemcitabine therapy in a patient with metastatic non-small-cell lung cancer. PMID- 17405820 TI - Patients' use and perception of medication information leaflets. AB - BACKGROUND: Federal law mandates that at least 95% of patients receive useful written information with new medications. Recent data concerning whether patients read such leaflets are lacking. OBJECTIVES: To evaluate whether patients read non manufacturer-developed leaflets and assess patients' opinions concerning the understandability and usefulness of these leaflets. METHODS: Patients were surveyed at 32 community pharmacies in the New York City metropolitan area. The main outcome measures were the percentage of patients who read the leaflet provided with new and refilled medications and the perceived understandability and usefulness of the leaflet. Descriptive statistics were used to analyze the results. RESULTS: A total of 307 patients were surveyed. For leaflets provided with new medications, 151 (49.2%), 65 (21.2%), 49 (16.0%), and 42 (13.7%) reported reading the leaflets always, often, seldom, or never, respectively. For refilled medications, 64 (21.6%), 41 (13.9%), 78 (26.4%), and 113 (38.2%) of the 296 respondents reported reading the leaflets always, often, seldom, or never, respectively. Of 267 patients who read the leaflets provided with new or refilled prescriptions at least seldom, 258 (96.6%) responded to the question concerning its understandability and 257 (96.3%) responded to the question concerning its usefulness. One hundred forty-five (56.2%), 89 (34.5%), 22 (8.5%), and 2 (0.8%) reported that the leaflet was very easy, somewhat easy, somewhat difficult, and very difficult to understand, respectively, and 164 (63.8%), 90 (35.0%), and 3 (1.2%) reported that the leaflet was very useful, somewhat useful, and not useful, respectively. CONCLUSIONS: Approximately two-thirds of the patients surveyed reported reading the leaflets provided with new medications at least often. The majority reported the leaflets to be useful and easy to understand. Pharmacists should advocate reading the leaflet and promote it as a useful resource. The leaflet should not replace the pharmacist's obligation to provide verbal counseling. PMID- 17405821 TI - Written versus oral recommendations made by pharmacy students during internal medicine rotations. AB - BACKGROUND: Pharmacy students use a variety of methods to communicate with physicians during clinical rotations regarding pharmacotherapy concerns. Documented acceptance rates for oral or written recommendations, when studied individually, range between 64% and 95%. OBJECTIVE: To compare the acceptance rates of written versus oral recommendations made by pharmacy students on internal medicine (IM) rotations. METHODS: Fourth-year pharmacy students completing an IM rotation made oral or written recommendations to physicians at a large, community-based medical center from November 2005 through April 2006 (excluding December). The types of recommendations and outcomes of the interventions were recorded using a data collection form. The primary endpoint was to determine differences in acceptance rates for written versus oral recommendations. Secondary endpoints included comparing the recommendation types and their corresponding acceptance rates. Additionally, the acceptance rates for evidence-based medicine (EBM) interventions were determined. RESULTS: A total of 625 recommendations were made by 10 pharmacy students during the 5 month study period; 47.5% of these were oral. A total of 82.8% of oral recommendations were accepted compared with 54.2% of written recommendations (p < 0.0001). Over 90% of the total recommendations were drug related. Overall, 68% of these recommendations were accepted. The major types of drug-related recommendations were indication for use (42.7%), inappropriate dose (17.2%), inappropriate route (11.3%), inappropriate drug (8.5%), and duplicate therapy (6.5%). The remaining types of interventions were laboratory related (6.4%) and requests for drug information (3.2%). A total of 227 (36.3%) recommendations were based on EBM guidelines, with an acceptance rate of 67.8%. CONCLUSIONS: Pharmacy student recommendations are well received by IM physicians. Oral recommendations are accepted at a statistically significantly higher percentage compared with written recommendations. High acceptance rates for recommendations may have the ability to positively impact patient care. PMID- 17405822 TI - Polyarteritis nodosa resistant to conventional treatment in a pediatric patient. AB - OBJECTIVE: To report the case of a child diagnosed with polyarteritis nodosa (PAN) that was unresponsive to conventional treatment alone but improved with the addition of iloprost and bosentan to her drug regimen. CASE SUMMARY: A 3-year-old girl who had been diagnosed with PAN was referred to our hospital from another region. With conventional treatment of high doses of a corticosteroid and cyclophosphamide, her condition resolved. Six months later, our patient had a relapse that required hospital admission. In this second hospital stay, some cutaneous lesions evolved into digital necrosis. Offlabel therapeutic alternatives, including a single dose (2 g/kg) of intravenous immunoglobulin (IVIG), intravenous iloprost 2 ng/kg/min over 6 h for 5 days and, approximately 4 wk later, oral bosentan 37.25 mg twice daily for 4 wk followed by 62.5 mg twice daily for 8 wk, were added to the conventional regimen to treat the serious cutaneous manifestations. Her fingers improved very slowly, and she was discharged on gradually tapered doses of oral corticosteroids, bosentan, and monthly pulsed injections of cyclophosphamide. The digital necrosis and other cutaneous lesions had resolved completely 6 months after the second discharge. DISCUSSION: The dosages of IVIG and iloprost were based on those used for PAN, Raynaud's phenomenon, and digital necrosis in children. The use of bosentan for vasculitis had not been reported in children before the treatment of our patient, so its dosage was based on that used to produce vasodilation in children with pulmonary hypertension. CONCLUSIONS: Digital necrosis and cutaneous manifestations not resolved with conventional PAN treatment improved within 5 days with iloprost and 12 weeks with bosentan. PMID- 17405823 TI - The role of selegiline in the treatment of negative symptoms associated with schizophrenia. AB - OBJECTIVE: To evaluate the role of selegiline in the treatment of negative symptoms associated with schizophrenia. DATA SOURCES: MEDLINE (1966-January 2007) and PsychINFO (1967-January 2007) were searched, using the terms schizophrenia, negative symptoms, and selegiline. A bibliographic search was conducted, as well. STUDY SELECTION AND DATA EXTRACTION: All English-language articles identified from the search were evaluated. All primary literature was included in the review. DATA SYNTHESIS: Based on its dopamine-enhancing property, selegiline has been studied as augmentation to antipsychotic therapy for the treatment of negative symptoms associated with schizophrenia. The efficacy of low-dose oral selegiline for the treatment of negative symptoms has been evaluated in 1 case report, 2 open-label trials, and 2 controlled trials. The case report and both open-label trials report improvement of negative symptoms associated with low dose oral selegiline. In 1 of the controlled trials, selegiline showed no difference in effect from that of placebo. These data are limited by small sample sizes. The largest controlled trial demonstrated a statistically significant difference between selegiline and placebo; however, the clinical significance is questionable, given that patients treated with selegiline were still experiencing marked negative symptoms at study completion. No comparative studies evaluating low-dose oral selegiline versus other augmentative treatment options for negative symptoms associated with schizophrenia exist at this time. CONCLUSIONS: Given the limitations of current literature, low-dose oral selegiline cannot be recommended for treatment of negative symptoms associated with schizophrenia. Additional controlled trials are needed to better delineate whether there is a role for selegiline in decreasing the burden of negative symptoms associated with schizophrenia. PMID- 17405824 TI - Hyponatremia: current treatment strategies and the role of vasopressin antagonists. AB - OBJECTIVE: Hyponatremia is a complex electrolyte disorder that results mainly from dysregulation of arginine vasopressin (AVP) by osmotic and nonosmotic mechanisms. Several populations, including the elderly, are at risk for the development of hyponatremia, and awareness of such risk factors can lead to an overall improvement in patient care. AVP receptor antagonists are promising new therapeutic options whose safety and efficacy have been clinically established for some forms of hyponatremia. DATA SOURCES: A search of MEDLINE (1967-March 2007) was conducted, using the search terms aquaporins, AVP, AVP receptor antagonists, conivaptan, hyponatremia, lixivaptan, and tolvaptan. Additionally, data were obtained from manufacturers' prescribing information. STUDY SELECTION AND DATA EXTRACTION: All articles identified from the reference search and data sources were evaluated. Material was included in this review if it was relevant to the pathophysiology and management of euvolemic and hypervolemic hyponatremia in hospitalized patients. DATA SYNTHESIS: A large percentage of hospitalized patients are found to have symptomatic or asymptomatic hyponatremia. One study reported that 24.5% of intensive care patients admitted over a 3 month period experienced hyponatremia at some time during their hospitalization. Conventional management techniques include water restriction, demeclocycline, lithium, and urea, which have demonstrated variable efficacy and toxicity. AVP receptor antagonists, whose safety and efficacy have been established in clinical trials, are providing new therapeutic options. CONCLUSIONS: AVP receptor antagonists appear to be safe and effective for the treatment of patients with hyponatremia. With conivaptan recently approved by the Food and Drug Administration for use in treating euvolemic and hypervolemic hyponatremia in hospitalized patients and lixivaptan and tolvaptan in the late stages of development, prudent use of these agents requires a thorough understanding of the clinical manifestations of hyponatremia to optimize therapeutic outcomes. PMID- 17405825 TI - The regulation of FSHbeta transcription by gonadal steroids: testosterone and estradiol modulation of the activin intracellular signaling pathway. AB - Recent reports suggest that androgens increase FSHbeta transcription directly via the androgen receptor and by modulating activin signaling. Estrogens may also regulate FSHbeta transcription in part through the activin system. Activin signaling can be regulated extracellularly via activin, inhibin, or follistatin (FS) or intracellularly via the Smad proteins. We determined the effects of androgen and estrogen on FSHbeta primary transcript (PT) concentrations in male and female rats, and we correlated those changes with pituitary: activin betaB mRNA, FS mRNA, the mRNAs for Smads2, -3, -4, and -7, and the phosphorylation (p) status of Smad2 and -3 proteins. In males, testosterone (T) increased FSHbeta PT two- to threefold between 3 and 24 h and was correlated with reduced FS mRNA, transient increases in Smad2, -4, and -7 mRNAs, and a six- to 10-fold increase in pSmad2, and activin betaB mRNA was unchanged. In females, T also increased FSHbeta PT twofold and pSmad2 threefold but had no effect on activin betaB, FS, or the Smad mRNAs. Androgen also increased Smad2 phosphorylation in gonadotrope derived alphaT3 cells. In contrast, estradiol had no effect on FSHbeta PT but transiently increased activin betaB mRNA and suppressed FS mRNA before increasing FS mRNA at 24 h and increased Smads2, -3, and -7 mRNAs and pSmad2 threefold. In conclusion, T acts on the pituitary to increase FSHbeta PT in both sexes and modulates FS mRNA, Smad mRNAs, and/or Smad2 phosphorylation. These findings suggest that T regulates FSHbeta transcription, in part, through modulation of various components of the activin-signaling system. PMID- 17405826 TI - Ghrelin and its unacylated isoform stimulate the growth of adrenocortical tumor cells via an anti-apoptotic pathway. AB - Ghrelin is expressed in normal human adrenocortical cells and induces their proliferation through growth hormone secretagogue receptor 1a (GHS-R1a). Consequently, it was of interest to us to determine whether acylated ghrelin and its predominant serum isoform, unacylated ghrelin, also act as factors for adrenocortical carcinoma cell growth. To examine a potential ghrelin-regulated system in adrenocortical tumors, we measured proliferative effects of acylated and unacylated ghrelin in the adrenocortical carcinoma cell lines SW-13 and NCI H295R. We also examined the expression of ghrelin, GHS-R1a, and corticotrophin releasing factor receptor 2 (CRF-R2). Acylated and unacylated ghrelin in the nanomolar range dose-dependently induced adrenocortical cell growth up to 200% of untreated controls, as measured by thymidine uptake and WST1 assay. The proliferative effects of acylated and unacylated ghrelin in SW-13 cells was blocked by [D-Lys(3)]growth hormone-releasing peptide 6 (GHRP6), but a CRF-R2 antagonist had no effect on unacylated ghrelin growth stimulation. Cell cycle analysis suggests that acylated and unacylated ghrelin suppress the sub G(0)/apoptotic fraction by up to 50%. Measurement of DNA fragmentation and caspase-3 and -7 activity in SW-13 cells confirmed that acylated and unacylated ghrelin suppress apoptotic rate. SW-13 cells express preproghrelin mRNA and secrete ghrelin, and [D-Lys(3)]GHRP6 suppresses their basal proliferation rate, strongly suggesting that ghrelin could act as an auto/paracrine growth factor. Acylated and unacylated ghrelin are potential auto/paracrine factors acting through an antiapoptotic pathway to stimulate adrenocortical tumor cell growth. Unacylated ghrelin-stimulated growth is suppressed by an antagonist of GHS-R1a, suggesting either that unacylated ghrelin is acylated before its action or that ghrelin, unacylated ghrelin, and [D-Lys(3)]GHRP-6 bind to a novel receptor in these cells. PMID- 17405827 TI - Synergism between psychosocial and metabolic stressors: impact on reproductive function in cynomolgus monkeys. AB - The role of energy imbalance versus psychosocial stress in the pathogenesis of female reproductive dysfunction characterized by anovulation and amenorrhea remains controversial. In women, functional hypothalamic amenorrhea can develop in the absence of significant weight loss, excessive exercise, or profound psychosocial disruption. We posited, therefore, that commonplace, seemingly minor stressors that alone would have minimal impact upon reproductive function might interact synergistically such that combinations of stressors would cause a greater impairment of the reproductive axis than any single stressor alone. We then developed a monkey model to test this hypothesis. Adult female cynomolgus monkeys with normal menstrual cycles were randomized into three experimental groups and studied over four menstrual cycles. The groups were: low-level psychosocial stress (i.e., moving to a new housing environment; Move, n = 8), moderate energy imbalance (Exercise + Diet, n = 9); and all stressors in combination (Move + Exercise + Diet, n = 10). Food intake, body weight, menstrual cyclicity, and reproductive hormones were assessed for two control menstrual cycles followed by two experimental cycles during which the monkeys experienced the stressors. Abnormal cycles were considered to be abnormally long or anovulatory cycles. Few abnormal cycles occurred in the Move group (1 of 8 monkeys) and in the Exercise + Diet group (1 of 9 monkeys). In contrast, 7 of 10 monkeys in the Move + Exercise + Diet group displayed at least one abnormal cycle (chi(2) = 9.61, P = 0.008). These findings suggest that infertility due to hypothalamic hypogonadism can result from the combination of commonplace, seemingly minor stressors that often escape clinical attention. PMID- 17405828 TI - 6-Fluoro-6-deoxy-D-glucose as a tracer of glucose transport. AB - Glucose transport rates are estimated noninvasively in physiological and pathological states by kinetic imaging using PET. The glucose analog most often used is (18)F-labeled 2FDG. Compared with glucose, 2FDG is poorly transported by intestine and kidney. We examined the possible use of 6FDG as a tracer of glucose transport. Lacking a hydroxyl at its 6th position, 6FDG cannot be phosphorylated as 2FDG is. Prior studies have shown that 6FDG competes with glucose for transport in yeast and is actively transported by intestine. Its uptake by muscle has been reported to be unresponsive to insulin, but that study is suspect. We found that insulin stimulated 6FDG uptake 1.6-fold in 3T3-L1 adipocytes and azide stimulated the uptake 3.7-fold in Clone 9 cells. Stimulations of the uptake of 3OMG, commonly used in transport assays, were similar, and the uptakes were inhibited by cyclochalasin B. Glucose transport is by GLUT1 and GLUT4 transporters in 3T3-L1 adipocyte and by the GLUT1 transporter in Clone 9 cells. Cytochalasin B inhibits those transporters. Rats were also imaged in vivo by PET using 6(18)FDG. There was no excretion of (18)F into the urinary bladder unless phlorizin, an inhibitor of active renal transport, was also injected. (18)F activity in brain, liver, and heart over the time of scanning reached a constant level, in keeping with the 6FDG being distributed in body water. In contrast, (18)F from 2(18)FDG was excreted in relatively large amounts into the bladder, and (18)F activity rose with time in heart and brain in accord with accumulation of 2(18)FDG-6-P in those organs. We conclude that 6FDG is actively transported by kidney as well as intestine and is insulin responsive. In trace quantity, it appears to be distributed in body water unchanged. These results provide support for its use as a valid tracer of glucose transport. PMID- 17405829 TI - Caffeine-induced Ca(2+) release increases AMPK-dependent glucose uptake in rodent soleus muscle. AB - Previous studies have proposed that caffeine-induced activation of glucose transport in skeletal muscle is independent of AMP-activated protein kinase (AMPK) because alpha-AMPK Thr172 phosphorylation was not increased by caffeine. However, our previous studies, as well as the present, show that AMPK phosphorylation measured in whole muscle lysate is not a good indicator of AMPK activation in rodent skeletal muscle. In lysates from incubated rat soleus muscle, a predominant model in previous caffeine-studies, both acetyl-CoA carboxylase-beta (ACCbeta) Ser221 and immunoprecipitated alpha(1)-AMPK activity increased with caffeine incubation, without changes in AMPK phosphorylation or immunoprecipitated alpha(2)-AMPK activity. This pattern was also observed in mouse soleus muscle, where only ACCbeta and alpha(1)-AMPK phosphorylation were increased following caffeine treatment. Preincubation with the selective CaMKK inhibitor STO-609 (5 microM), the CaM-competitive inhibitor KN-93 (10 microM), or the SR Ca(2+) release blocking agent dantrolene (10 microM) all inhibited ACCbeta phosphorylation and alpha(1)-AMPK phosphorylation, suggesting that SR Ca(2+) release may work through a CaMKK-AMPK pathway. Caffeine-stimulated 2-deoxyglucose (2DG) uptake reflected the AMPK activation pattern, being increased with caffeine and inhibited by STO-609, KN-93, or dantrolene. The inhibition of 2DG uptake is likely causally linked to AMPK activation, since muscle-specific expression of a kinase-dead AMPK construct greatly reduced caffeine-stimulated 2DG uptake in mouse soleus. We conclude that a SR Ca(2+)-activated CaMKK may control alpha(1) AMPK activation and be necessary for caffeine-stimulated glucose uptake in mouse soleus muscle. PMID- 17405830 TI - Antidiabetic sulfonylurea stimulates insulin secretion independently of plasma membrane KATP channels. AB - Understanding mechanisms by which glibenclamide stimulates insulin release is important, particularly given recent promising treatment by glibenclamide of permanent neonatal diabetic subjects. Antidiabetic sulfonylureas are thought to stimulate insulin secretion solely by inhibiting their high-affinity ATP sensitive potassium (K(ATP)) channel receptors at the plasma membrane of beta cells. This normally occurs during glucose stimulation, where ATP inhibition of plasmalemmal K(ATP) channels leads to voltage activation of L-type calcium channels for rapidly switching on and off calcium influx, governing the duration of insulin secretion. However, growing evidence indicates that sulfonylureas, including glibenclamide, have additional K(ATP) channel receptors within beta cells at insulin granules. We tested nonpermeabilized beta-cells in mouse islets for glibenclamide-stimulated insulin secretion mediated by granule-localized K(ATP) channels by using conditions that bypass glibenclamide action on plasmalemmal K(ATP) channels. High-potassium stimulation evoked a sustained rise in beta-cell calcium level but a transient rise in insulin secretion. With continued high-potassium depolarization, addition of glibenclamide dramatically enhanced insulin secretion without affecting calcium. These findings support the hypothesis that glibenclamide, or an increased ATP/ADP ratio, stimulates insulin secretion in part by binding at granule-localized K(ATP) channels that functionally contribute to sustained second-phase insulin secretion. PMID- 17405831 TI - The human reticulocyte transcriptome. AB - RNA from circulating blood reticulocytes was utilized to provide a robust description of genes transcribed at the final stages of erythroblast maturation. After depletion of leukocytes and platelets, Affymetrix HG-U133 arrays were hybridized with probe generated from the reticulocyte total RNA (blood obtained from 14 umbilical cords and 14 healthy adult humans). Among the cord and adult reticulocyte profiles, 698 probe sets (488 named genes) were detected in each of the 28 samples. Among the highly expressed genes, promoter analyses revealed a subset of transcription factor binding motifs encoded at higher than expected frequencies including the hypoxia-related arylhydrocarbon receptor repressor family. Over 100 probe sets demonstrated differential expression between the cord and adult reticulocyte samples. For verification, the array expression patterns for 21 genes were confirmed by real-time PCR (correlation coefficient 0.98). Only four transcripts (MAP17, FLJ32009, ARRB2, and FLJ27365) were identified as being upregulated in the adult blood transcriptome. Further analysis revealed that the lipid-regulating protein MAP17 was present in the membrane fraction of adult erythrocytes, but not detected in cord blood erythrocytes. Combined with other clinical and experimental data, these reticulocyte transcriptome profiles should be useful to better understand the molecular bases of terminal erythroid differentiation, hemoglobin switching, iron metabolism and malarial pathogenesis. PMID- 17405832 TI - Is progesterone elevation on the day of human chorionic gonadotrophin administration associated with the probability of pregnancy in in vitro fertilization? A systematic review and meta-analysis. AB - The role of progesterone elevation on in vitro fertilization (IVF) outcome has remained a debatable issue for several years. The aim of this systematic review and meta-analysis was to evaluate whether progesterone elevation on the day of human chorionic gonadotrophin (hCG) administration is associated with the probability of pregnancy. Eligible studies were considered those in which patients did not participate more than once. A literature search in MEDLINE, EMBASE and CENTRAL identified 12 eligible studies, 10 of which were retrospective. The majority (n = 10) of these studies did not detect a statistically significant association between progesterone elevation and the probability of pregnancy. Meta-analysis was performed only for the studies (n = 5) that provided data on clinical pregnancy per patient reaching hCG administration for final oocyte maturation. No statistically significant association between progesterone elevation and the probability of clinical pregnancy was detected (Odds ratio: 0.75, 95% confidence interval 0.53-1.06; P = 0.10). This finding persisted in the sensitivity analyses performed, which excluded the studies that did not report clearly that measurement of progesterone did not affect patients' management and those that did not report definition of clinical pregnancy. In addition, subgroup analyses were conducted on the basis of type of gonadotrophin-releasing hormone GnRH analogue used and on the value of serum threshold used to classify patients in those with or without progesterone elevation. These analyses, however, did not materially change the results obtained. In conclusion, the best available evidence does not support an association between progesterone elevation on the day of hCG administration and the probability of clinical pregnancy in women undergoing ovarian stimulation with GnRH analogues and gonadotrophins for IVF. PMID- 17405833 TI - Interferon-gamma and interleukin-4 gene polymorphisms in Caucasian idiopathic inflammatory myopathy patients in UK. AB - OBJECTIVE: To determine whether interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) genes confer susceptibility for the idiopathic inflammatory myopathies (IIMs). METHODS: A large cross-sectional study of UK caucasian adults with polymyositis (PM, n = 101), dermatomyositis (DM, n = 94) and myositis overlapping with a connective tissue disease (myositis/CTD-overlap, n = 70) was completed. 177 ethnically matched controls were available for comparison. Single-nucleotide polymorphisms (SNPs) within intronic regions coding for IL-4, IFN-gamma and a microsatellite marker within intron 1 of the IFN-gamma gene were typed. RESULTS: Strong linkage disequilibrium was present between SNPs in each gene. In the IFN gamma gene, a weak allelic association was observed in PM versus controls at rs1861493 (odds ratio (OR) 1.6, 95% confidence interval (CI) 1.03 to 2.4). The microsatellite IFN-gamma CA(14) allele was associated with risk for IIMs overall (OR 3.3, 95% CI 1.4 to 7.8), the strongest association being observed within the anti-U1-ribonucleoprotein (RNP) group (OR 6.0, 95% CI 1.5 to 23.1), and persisting after adjustment for known myositis human leucocyte antigen (HLA) class II associations. CONCLUSIONS: Genetic markers in the IFN-gamma gene demonstrate significant allelic associations with the IIMs in a UK Caucasian population. The SNPs tested in this study within the region coding for IL-4 fail to show significant associations with susceptibility to IIM disease. PMID- 17405834 TI - Patient preferences for treatment: report from a randomised comparison of treatment strategies in early rheumatoid arthritis (BeSt trial). AB - OBJECTIVE: To determine treatment preferences among patients with recent onset rheumatoid arthritis participating in a randomised controlled trial comparing four therapeutic strategies. METHODS: A questionnaire was sent to all 508 participants of the BeSt trial, treated for an average of 2.2 years with either sequential monotherapy (group 1), step-up combination therapy (group 2), initial combination therapy with tapered high-dose prednisone (group 3), or initial combination therapy with infliximab (group 4). Treatment adjustments were made every 3 months to achieve low disease activity (DAS < or =2.4). The questionnaire explored patients' preferences or dislikes for the initial therapy. RESULTS: In total, 440 patients (87%) completed the questionnaire. Despite virtually equal study outcomes at 2 years, more patients in group 4 reported much or very much improvement of general health: 50%, 56%, 46% and 74% in groups 1-4, respectively (overall, P<0.001). Almost half of the patients expressed no preference or aversion for a particular treatment group, 33% had hoped for assignment to group 4 and 38% had hoped against assignment to group 3. This negative perception was much less prominent in patients actually in group 3. Nevertheless, 50% of patients in group 3 disliked having to take prednisone, while only 8% in group 4 disliked going to the hospital for intravenous treatment. CONCLUSIONS: Within the limitations of our retrospective study, patients clearly preferred initial combination therapy with infliximab and disliked taking prednisone. After actual exposure, this preference remained, but the perception of prednisone improved. Patient perceptions need to be addressed when administering treatment. PMID- 17405835 TI - Calcitonin measurement in wash-out fluid from fine needle aspiration of neck masses in patients with primary and metastatic medullary thyroid carcinoma. AB - OBJECTIVE: The aim of the study was to evaluate the usefulness of calcitonin (CT) assay in fine-needle aspiration biopsy (FNAB) wash-out fluid alone or combined with cytology in the presurgical study of medullary thyroid carcinoma (MTC) patients with thyroid nodules (TNs) and of suspicious neck MTC recurrences/metastases. SUBJECTS AND METHODS: A total of 36 ultrasound-guided FNABs were performed in neck masses from 23 patients with borderline or high basal and pentagastrin-stimulated serum CT. Cytology and CT-FNAB were performed on a total of 18 TNs and three neck lymph nodes (LNs) from 12 patients examined before thyroidectomy, and on six suspicious local recurrences (LRs) and nine LNs from nine totally thyroidectomized MTC patients. On the basis of CT-FNAB values found in 15 non-MTC lesions, CT-FNAB more than 36 pg/ml was considered as indicative of MTC. RESULTS: All 21 positive CT-FNAB lesions (10 TNs, six LNs, and five LRs), 13 with positive cytology, were confirmed as MTC at histology. Of the 15 negative CT-FNAB suspicious masses (eight TNs, six LNs, and one LR), five displayed a benign lesion at histology. The remaining 10 cases, all with benign cytology, were not operated on, and no evidence of MTC was detected at follow-up. CT-FNAB reached 100% sensitivity and specificity for MTC, while cytology displayed 61.9% sensitivity and 80% specificity. CONCLUSIONS: Ultrasound-guided CT-FNAB was the best tool to identify primary MTC and LRs/node metastases in MTC operated subjects. This may have important implications in the management of MTC. PMID- 17405836 TI - Tripartite control of growth hormone secretion in women during controlled estradiol repletion. AB - CONTEXT: Studies of how aging attenuates GH secretion are confounded by differences in sex-steroid milieus, abdominal visceral fat mass (AVF), and IGF-I concentrations and limited in interpretability by the use of pharmacological doses of secretagogues. HYPOTHESIS: In a controlled estrogenic milieu, near physiological secretagogue drive will unmask distinct influences of age, AVF, and IGF-I on GH secretion. LOCATION: The study was conducted at an academic medical center. SUBJECTS: Subjects included 10 healthy pre- (PRE) and 10 postmenopausal (POST) women. PROCEDURE: In a defined estradiol (E(2)) milieu, we compared GH secretion after submaximal stimulation with GH-releasing peptide (GHRP)-2 (ghrelin analog), GHRH, and l-arginine (an inhibitor of somatostatin outflow). ANALYSIS: We related GH responses to age stratum (dichotomous variable) and AVF and IGF-I concentrations (continuous variables). RESULTS: In the face of comparable concentrations of E(2), testosterone, and SHBG: 1) age (P < 0.001) and secretagogue type (P < 0.001) independently determined GH secretion; 2) GH responses in POST subjects were only 26-33% of those in PRE (P < or = 0.002) across all secretagogues; 3) POST women lost the PRE order of secretagogue potency (GHRP-2 > GHRH = l-arginine); and 4) in the combined cohorts, higher AVF predicted reduced l-arginine-stimulated GH secretion (R(2) = 0.46, P = 0.0013), whereas higher IGF-I concentrations forecast increased GHRP-2 and GHRH drive (R(2) > or = 0.52, P < or = 0.013). CONCLUSION: A paradigm of near-physiological secretagogue drive in an E(2)-clamped milieu unmasks tripartite deficits in peptide-signaling pathways in healthy POST, compared with PRE, women. Post hoc analyses indicate that both greater visceral adiposity and lower IGF-I concentrations mark this triple regulatory defect. PMID- 17405837 TI - Pharmacokinetics of recombinant methionyl human leptin after subcutaneous administration: variation of concentration-dependent parameters according to assay. AB - CONTEXT: Recombinant human leptin (r-metHuLeptin) has demonstrated efficacy in improving hormonal and metabolic parameters in leptin-deficient states, but pharmacokinetic parameters after sc administration have not yet been published. In addition, the effect of potential variability across different leptin assays on concentration-dependent pharmacokinetic parameters remains unknown. OBJECTIVE: The objective of the study was to characterize pharmacokinetic parameters after sc r-metHuLeptin administration using three commercially available leptin assays (Linco, Diagnostic Systems Laboratories, and Alpco). DESIGN, SETTING, PATIENTS, AND INTERVENTION: We analyzed pharmacokinetic profiles in five lean and five obese men after sc administration of physiological (0.01 mg/kg) and pharmacological (0.3 mg/kg) doses of r-metHuLeptin. MAIN OUTCOME MEASURES: Leptin pharmacokinetic parameters were measured. RESULTS: Measurement of leptin produced typical pharmacokinetic profiles in all assays with time to maximal concentration and half-life of approximately 3 h. Diagnostic Systems Laboratories consistently measured leptin higher than Linco, with Alpco measuring intermediate between or similar to Linco. There was high correlation among assays (R(2) ranging from 0.89 to 0.98, all P < 0.01). Concentration-dependent parameters such as maximal concentration, area under the curve, and clearance were significantly different among assays, whereas concentration-independent parameters such as time to maximal concentration and half-life were generally not different. CONCLUSIONS: We report novel data on leptin pharmacokinetic parameters after sc administration, which will be relevant for the future therapeutic use of r-metHuLeptin. Although commercially available assays demonstrated high correlation, they can provide substantially different measures of leptin levels. This demonstrates the importance of standardizing leptin assays for diagnosing patients with relative leptin deficiency, determining appropriate doses of r-metHuLeptin for administration, and monitoring response to therapy. PMID- 17405838 TI - Abdominal fat quantity and distribution in women with polycystic ovary syndrome and extent of its relation to insulin resistance. AB - CONTEXT: Increased abdominal fat has been linked to insulin resistance and increased cardiovascular risk. Because many patients with polycystic ovary syndrome (PCOS) present abdominal obesity, it may be the cause of insulin resistance in this disorder. SETTING: Fat quantity and distribution were evaluated by dual x-ray absorptiometry at the Departments of Clinical Medicine at the University of Palermo and the University of Naples, Italy. PATIENTS: A total of 110 patients with PCOS and 112 weight-matched controls were studied. Anthropometric data, blood glucose, serum insulin, and testosterone were evaluated. Total, trunk, and central abdominal fat quantity were measured by total-body dual x-ray absorptiometry. RESULTS: Compared with weight-matched controls, patients with PCOS had similar quantity of total and trunk fat but higher quantity of central abdominal fat. This difference was not observed when comparing obese PCOS and obese controls but depended on differences between overweight and normoweight patients and controls. All obese subjects, independently of having PCOS or not, had increased central abdominal fat. The same parameter was increased in 71% of overweight PCOS, 50% of overweight controls, and 30% of normoweight PCOS patients. PCOS patients with increased central abdominal fat had significantly higher (P < 0.01) insulin levels and significantly reduced (P < 0.01) insulin sensitivity than controls with similar quantities of central abdominal fat. Overweight PCOS patients with normal abdominal fat had significantly higher (P < 0.05) insulin levels and significantly reduced (P < 0.05) insulin sensitivity than overweight controls with normal abdominal fat. CONCLUSIONS: Most obese subjects, independent of being affected by PCOS, have an abdominal form of obesity. However, abdominal fat excess may not be the only determinant of insulin resistance in PCOS. PMID- 17405839 TI - Genetic variations at the endocannabinoid type 1 receptor gene (CNR1) are associated with obesity phenotypes in men. AB - CONTEXT: The endocannabinoid system modulates food intake and body weight in animal models. Treatment with the cannabinoid type 1 receptor blocker, rimonabant, reduces body weight in obese individuals. OBJECTIVE: The aim of this study was to determine whether single nucleotide polymorphisms of the gene encoding cannabinoid type 1 receptor, CNR1, are associated with body fat mass and distribution in two independent samples of white European adult men. DESIGN, SETTING, AND PARTICIPANTS: The 3813A/G and 4895A/G single nucleotide polymorphisms at the exon 4 of CNR1 were genotyped in 930 participants to the Olivetti Prospective Heart Study (OPHS) in Southern Italy and in 216 participants to the Wandsworth Heart and Stroke Study in the United Kingdom. Retrospective analysis was also performed on an OPHS subsample (n = 360) for which anthropometric data from 1987 and 1994-1995 examinations were available. MAIN OUTCOME MEASURES: CNR1 genotypes and anthropometric measures of body fat distribution were determined. RESULTS: In the OPHS study, the 3813G allele was associated with increased subscapular skinfold thickness (24.2 +/- 9.1 vs. 22.8 +/- 7.7 mm; P = 0.031) and waist circumference (WC) (99.1 +/- 8.8 vs. 97.7 +/- 8.8 cm; P = 0.050). No association was observed with 4895A/G variant. Haplotype analysis confirmed that the unique haplotype carrying the 3813G was associated with increased WC and subscapular skinfold thickness. Similar results were observed in the OPHS retrospective subsample and the Wandsworth Heart and Stroke Study sample. In the latter, the 3813G was associated with increased WC (96.8 +/- 11.3 vs. 91.6 +/- 10.4 cm; P = 0.006). CONCLUSIONS: Genetic variants at CNR1 are associated with obesity-related phenotypes in men. The detection of polymorphic variants in genes involved in the process of fat accumulation may help identify specific targets for pharmacological treatment of obesity and related metabolic abnormalities. PMID- 17405840 TI - Evidence of prolonged orocecal transit time and small intestinal bacterial overgrowth in acromegalic patients. AB - CONTEXT: Gastrointestinal abnormalities in acromegaly include dolichomegacolon, slow colonic transit, and increased prevalence of colonic polyps. Conversely, no data are available on the small intestine. OBJECTIVE: The aim of the study was to investigate the orocecal transit time (OCTT) and the presence of small intestinal bacterial overgrowth (SIBO). PATIENTS: A total of 41 acromegalic patients and 30 sex- and age-matched control subjects entered the study. Acromegalic patients were classified according to the medical treatment with somatostatin analogs as "treated" (n = 22) and "untreated" (n = 19), whereas according to the disease control, as "controlled" (n = 17), "uncontrolled" (n = 10), and "partially controlled" (n = 14). Patients and controls completed a questionnaire and underwent a standardized 10-g lactulose hydrogen breath test to determine the OCTT and presence of SIBO. SIBO-positive patients underwent eradication with rifaximine. RESULTS: An increased prevalence of SIBO (18 of 41 vs. 1 of 30; P < 0.0001) and a significantly delayed OCTT (169.53 +/- 8.15 vs. 107.25 +/- 6.56 min; P < 0.0001) were evidenced in patients compared with controls. No significant statistical differences were found between "treated" or "untreated" patients positive for SIBO or between "controlled," "partially controlled," and "uncontrolled" patients. OCTT was significantly delayed in "treated" vs. "untreated" patients (183.21 +/- 9.01 and 158.89 +/- 6.38, respectively; P = 0.02) and in patients compared with controls (105.75 +/- 6.34; P < 0.0001). Rifaximine eradicated SIBO in more than 50% of patients who underwent treatment. CONCLUSIONS: These data demonstrate for the first time that SIBO occurs more frequently in acromegalic patients, however, it can be successfully treated by a specific antibiotic. Medical therapy with somatostatin analogs does not affect SIBO prevalence. OCTT resulted significantly prolonged in both "treated" and "untreated" patients, suggesting that acromegaly determines per se an impairment of the intestinal motility. Indeed, disease control seems irrelevant on the delayed OCTT, suggesting that this alteration might be an irreversible complication of acromegaly, probably related to an autonomic intestinal disorder, as we have previously demonstrated at the cardiac level. PMID- 17405841 TI - The expression of NAD(P)H:quinone oxidoreductase 1 is high in human adipose tissue, reduced by weight loss, and correlates with adiposity, insulin sensitivity, and markers of liver dysfunction. AB - CONTEXT: We have previously identified nicotinamide adenine dinucleotide phosphate:quinone oxidoreductase 1 (NQO1), an enzyme involved in the protection against oxidative stress, as a gene predominantly expressed in human adipocytes. Studies in mice deficient in NQO1 activity suggest that NQO1 may also play an important role in metabolism. OBJECTIVE: The aim of this study was to explore the expression and regulation of NQO1 in human adipose tissue (AT) and isolated adipocytes. PATIENTS AND RESULTS: The high expression of NQO1 in adipocytes was verified in human adipocytes and AT by real-time PCR. DNA microarray analysis showed that NQO1 was expressed at higher levels in large compared with small adipocytes, isolated from the same fat biopsy. Furthermore, NQO1 mRNA levels were positively correlated with adipocyte size (n = 7; P < 0.002). During an 18-wk diet regime (n = 24; mean weight loss 27 kg), the NQO1 expression in human sc AT was down-regulated (P < 0.0001), and mRNA levels correlated with body mass index (P = 0.0005), sc, and total abdominal AT areas, as determined by computerized tomography (P < 0.0001, both) and metabolic parameters. NQO1 mRNA levels were also positively correlated with aspartate aminotransferase (P = 0.0028) and alanine aminotransferase (P = 0.0219), markers known to be associated with severity of hepatic steatosis. CONCLUSIONS: NQO1 is highly expressed in human AT, particularly in large adipocytes. AT NQO1 expression is reduced during diet induced weight loss, and the expression levels positively correlate with adiposity, glucose tolerance, and markers of liver dysfunction. Together, these findings indicate a role for NQO1 in the metabolic complications of human obesity. PMID- 17405842 TI - Identification and developmental changes of aromatase and estrogen receptor expression in prepubertal and pubertal human adrenal tissues. AB - CONTEXT: The mechanisms of postnatal adrenal zonation remain unclear. OBJECTIVE: To provide a clue for a possible role of estrogens in adrenarche, we studied the expression of estrogen receptor (ER)alpha, ERbeta, G protein-coupled receptor (GPR)30, and cP450aromatase (cP450arom) in human adrenal tissue. DESIGN: Human adrenal tissue was collected from three postnatal age groups (Grs): Gr 1, younger than 3 months (n = 12), fetal zone involution; Gr 2, 3 months to 6 yr (n = 17), pre-adrenarche; and Gr 3, older than 6-20 yr (n = 12), post-adrenarche period. RESULTS: ERbeta mRNA in Grs 1 and 3 was higher than in Gr 2 (P < 0.05). By immunohistochemistry and laser capture microdissection followed by RT-PCR, ERbeta was expressed in zona reticularis and fetal zone, GPR30 in zona glomerulosa (ZG) and adrenal medulla, while ERalpha mRNA and protein were undetectable. cP450arom mRNA in Gr 3 was higher than in Grs 1 and 2 (P < 0.05), and localized to ZG and adrenal medulla by laser capture microdissection. cP450arom Immunoreactivity was observed in adrenal medulla in the three Grs and in subcapsular ZG of Gr 3. Double-immunofluorescence studies revealed that cP450arom and chromogranin A only colocalize in adrenal medulla of subjects younger than 18 months. In these samples, exon 1.b-derived transcript was 3.5-fold higher, while exon 1.a-, 1.c-, and 1.d-derived transcripts were 3.3-, 1.9-, and 1.7-fold lower, respectively, than in subjects older than 6 yr. CONCLUSIONS: Our results suggest that estrogens produced locally in adrenal medulla would play a role in zona reticularis functional differentiation through ERbeta. The cP450arom and GPR30 expression in subcapsular ZG, colocalizing with a high-cell proliferation index, previously reported, suggests a local GPR30-dependent estrogen action in proliferation and migration of progenitor adrenal cells. PMID- 17405843 TI - Epigenetic defects of GNAS in patients with pseudohypoparathyroidism and mild features of Albright's hereditary osteodystrophy. AB - CONTEXT: Several endocrine disorders that share resistance to PTH are grouped under the term pseudohypoparathyroidism (PHP). PHP type I, associated with blunted PTH-induced nephrogenous cAMP formation and phosphate excretion, is subdivided according to the presence or absence of additional endocrine abnormalities, Albright's hereditary osteodystrophy (AHO), and reduced Gsalpha activity caused by GNAS mutations. OBJECTIVE: We sought to identify the molecular defect in four unrelated patients who were thought to have PHP-Ia because of PTH and TSH resistance and mild AHO features. METHODS: Gsalpha activity and mutation analysis, and assessment of GNAS haplotype, methylation, and gene expression were performed for probands and family members. RESULTS: Two patients showed modest decreases in erythrocyte Gsalpha activity. Instead of Gsalpha point mutations, however, all four patients showed methylation defects of the GNAS locus, a feature previously described only for PHP-Ib. Furthermore, one patient with an isolated loss of GNAS exon A/B methylation had the 3-kb STX16 deletion frequently identified in PHP-Ib patients. In all but one of the remaining patients, haplotype analysis excluded large deletions or uniparental disomy as the cause of the observed methylation changes. CONCLUSIONS: Our investigations indicate that an overlap may exist between molecular and clinical features of PHP-Ia and PHP Ib. No current mechanisms can explain the AHO-like features of our patients, some of which may not be linked to GNAS. Therefore, patients with hormone resistance and AHO-like features in whom coding Gsalpha mutations have been excluded should be evaluated for epigenetic alterations within GNAS. PMID- 17405844 TI - Bilateral adrenal infiltration in Erdheim-Chester disease. Report of seven cases and literature review. AB - CONTEXT: Erdheim-Chester disease (ECD) is a rare non-Langerhans form of histiocytosis characterized by xanthomatous tissue infiltration with CD68(+) CD1a(-) foamy histiocytes. Endocrine problems, such as diabetes insipidus and hypogonadotropic hypogonadism, frequently occur in ECD, but bilateral adrenal infiltration has rarely been reported in this disease. OBJECTIVE: The aim of the study was to describe adrenal and periadrenal infiltration in ECD. PATIENTS: A total of 22 patients with ECD undergoing systematic computed tomography to search for signs of adrenal enlargement participated in the study. RESULTS: Of the 22 patients with ECD, seven (31.8%) displayed adrenal infiltration on computed tomography. In one case, autopsy confirmed that the adrenal enlargement was due to foamy histiocyte infiltration in the adrenal glands. Possible adrenal insufficiency was assessed in five of the seven patients. One developed signs of adrenal insufficiency, which was confirmed by adrenocorticotropin stimulation tests. Adrenal involvement was reported in only 15 of the 240 ECD cases published up to May 2006. This frequency is significantly lower than that in our series (P = 0.0008; Fisher's exact test). CONCLUSION: Physicians should be aware of ECD as a possible cause of morphological changes in adrenal size and infiltration. PMID- 17405845 TI - Hypoglycemia due to an insulin binding antibody in a patient with an IgA-kappa myeloma. AB - CONTEXT: Autoantibodies to insulin have been described to cause spontaneous hypoglycemia in nondiabetic subjects. There have been occasional reports of spontaneous hypoglycemia due to monoclonal anti-insulin antibodies. We present the first report of a patient with an IgA-kappa myeloma in whom frequent hypoglycemia resulted from the ability of the monoclonal IgA-kappa to bind insulin. OBJECTIVES: The aim of this study was to describe the occurrence of profound hypoglycemia in a patient with IgA-kappa myeloma, characterize biochemically the nature of the IgA:insulin complex present, and place this case in the context of the published literature on hypoglycemia resulting from autoantibodies to insulin. DESIGN: A case study was performed. PATIENTS: A single case of profound hypoglycemia associated with IgA-kappa myeloma was studied. INTERVENTION: There were no interventions. MAIN OUTCOME MEASURES: A case study was performed. RESULTS: Polyethylene glycol precipitation and gel filtration chromatography were used to demonstrate high-molecular weight insulin immunoreactivity in the patient's plasma. This was characterized as an insulin binding IgA-kappa paraprotein present at 4200 mg/dl (42 g/liter) with a relatively high insulin dissociation constant of 0.32 microm/liter using radiolabelled insulin binding studies. CONCLUSIONS: We present the first case of hypoglycemia due to IgA binding insulin antibodies in a patient with an IgA-kappa paraprotein myeloma. The hypoglycemia was associated with high-plasma insulin levels and relatively low C-peptide levels. A plausible mechanism for the hypoglycemia is the delayed clearance of insulin. This case broadens the spectrum of monoclonal gammopathies that have been associated with anti-insulin reactivity and spontaneous hypoglycemia. PMID- 17405846 TI - Plasma levels and adipose tissue messenger ribonucleic acid expression of retinol binding protein 4 are reduced during calorie restriction in obese subjects but are not related to diet-induced changes in insulin sensitivity. AB - CONTEXT: Retinol-binding protein 4 (RBP4) may play a role in the development of insulin resistance. OBJECTIVE: We investigated whether RBP4 adipose tissue mRNA expression and plasma level are related to insulin sensitivity during a diet induced weight loss. DESIGN, SETTING, PATIENTS, AND INTERVENTION: Obese women followed a dietary intervention composed of a 4-wk very low-calorie diet (VLCD), a 2-month low-calorie diet, and 3-4 months of a weight maintenance (WM) phase. MAIN OUTCOME MEASURES: Clinical investigation was performed before and at the end of each phase. Insulin sensitivity was assessed with the euglycemic hyperinsulinemic clamp. Adipose tissue mRNA and plasma levels of RBP4 were determined using reverse transcription-quantitative PCR and ELISA, respectively. RESULTS: Weight and fat mass decreased during VLCD and were stabilized during WM. Glucose disposal rate increased during VLCD and remained elevated thereafter. Plasma levels of RBP4 decreased after VLCD and, although increasing at subsequent phases, remained lower than prediet values. Adipose tissue mRNA levels were diminished after VLCD, and increased during low-calorie diet and WM to reach basal values. Basal RBP4 levels or diet-induced variations of RBP4 were not different in lean women and two groups of obese women with high- and low-insulin sensitivity. CONCLUSIONS: Severe calorie restriction promotes a reduction in adipose tissue and plasma levels of RBP4. The study does not bring evidence for a role for RBP4 in the regulation of diet-induced changes in insulin sensitivity. PMID- 17405847 TI - Primary growth hormone (GH) insensitivity and insulin-like growth factor deficiency caused by novel compound heterozygous mutations of the GH receptor gene: genetic and functional studies of simple and compound heterozygous states. AB - CONTEXT: Primary GH insensitivity (GHI) or Laron syndrome, caused by mutations of the GH receptor (GHR) gene, has a clinical phenotype of postnatal growth failure associated with normal elevated serum concentrations of GH and low serum levels of IGF-I. OBJECTIVE: We investigated the clinical and biochemical implications of molecular defects in the GHR gene in an Austrian family with two daughters who were GHI. PATIENTS: Patient 1 [height, -4.8 sd score (SDS)] and patient 2 (height, -5.0 SDS) had elevated circulating levels of GH, low-normal levels of GH binding protein, and abnormally low IGF-I (-5.0 SDS and -2.6 SDS, respectively) and IGF-binding protein-3 (-2.6 SDS and -2.0 SDS, respectively). RESULTS: Both patients carry novel compound, missense, heterozygous GHR mutations, C94S and H150Q. In vitro reconstitution experiments demonstrated that whereas each of the mutants could be stably expressed, GHR(C94S) lost its affinity for GH and could neither activate signal transducer and activator of transcription (STAT)-5b nor drive STAT5b-dependent gene transcription in response to GH (1-100 ng/ml). GHR(H150Q) showed normal affinity for GH but impaired capacity for signal transduction. The compound heterozygote and C94S heterozygote, but not the H150Q heterozygote, showed significant deficiency in activating GH-induced gene expression, corroborating diminished GH-induced STAT5b activation in fibroblasts carrying GHR(C94S) as either a compound heterozygote (in the patients) or a simple heterozygote (in one parent). CONCLUSIONS: Each of the compound heterozygous mutations contributed additively to the pathological condition seen in the patients, and the more detrimental of the two mutations, C94S, may cause (partial) primary GHI, even in a heterozygous state. PMID- 17405848 TI - Effect of an estrogen receptor-alpha intron 4 polymorphism on fat mass in 11-year old children. AB - CONTEXT: Polymorphisms in the ESR1 gene encoding estrogen receptor (ER)-alpha may be associated with fat mass in adults. OBJECTIVES: The objective of the study was to establish whether ESR1 polymorphisms influence fat mass in childhood. DESIGN: This was a cross-sectional analysis after genotyping of rs9340799, rs2234693, and rs7757956 ESR1 polymorphisms. SETTING: The Avon Longitudinal Study of Parents and Children (ALSPAC) was a population-based prospective study. PARTICIPANTS: Participants included 3097 11-yr-old children with results for ESR1 genotyping, puberty measures, and dual-energy x-ray absorptiometry results. OUTCOMES: Relationships between ESR1 polymorphisms and indices of body composition were measured. RESULTS: The rs7757956 polymorphism was associated with fat mass (P = 0.002). Total body fat mass (adjusted for height) was reduced by 6% in children with TA/AA genotypes, and risk of being overweight (> or =85th centile of fat mass) was decreased by 20%. This genetic effect appeared to interact with puberty in girls (P = 0.05 for interaction): in those with the TT genotype, total body fat mass (adjusted for height) was 18% higher in Tanner stages 3-5 vs. stages 1 2; the equivalent difference was 7% in those with TA/AA genotypes. Furthermore, the risk of being overweight was 36% lower in girls with TA/AA genotypes in Tanner stages 3-5, but no reduction was seen in those in stages 1-2. Neither rs9340799 nor rs2234693 polymorphisms were associated with body composition measures. CONCLUSIONS: Fat mass in 11-yr-old children was related to the rs7757956 ESR1 polymorphism. This association was strongest in girls in more advanced puberty, in whom the risk of being overweight was reduced by 36% in those with the TA/AA genotype. PMID- 17405849 TI - Predictive value and clinical impact of Basal follicle-stimulating hormone in subfertile, ovulatory women. AB - CONTEXT: Basal FSH is a marker for ovarian reserve. OBJECTIVES: The objective of the study was to investigate the predictive value of basal FSH on spontaneous ongoing pregnancy in subfertile ovulatory women. DESIGN: This was a prospective cohort study. SETTING: The study was conducted in 19 fertility centers in The Netherlands. PARTICIPANTS: Subfertile ovulatory women without two-sided tubal pathology and in whom the man had normal sperm parameters (total motile count > or = 3 x10(6)) participated in the study. INTERVENTIONS: Interventions included a fertility work-up, including a basal FSH measurement on cycle d 3. MAIN OUTCOME MEASURES: Spontaneous ongoing pregnancy was measured. RESULTS: We included 3519 consecutive couples of which 562 (16%) had a spontaneous ongoing pregnancy within 1 yr. Basal FSH levels of 8 IU/liter or higher were associated with a decreased probability of spontaneous ongoing pregnancy [hazard ratio (HR) 0.93/IU.liter (95% confidence interval [CI] 0.87-0.98)]. In a multivariable analysis, female age (HR 0.97/yr, 95% CI 0.95-0.99), cycle length (HR 0.96/d, 95% CI 0.93-1.0), and FSH levels 8 IU/liter or greater (HR 0.93/IU.liter, 95% CI 0.87-0.99) were strong negative predictors for spontaneous ongoing pregnancy. Addition of FSH to a prediction model based on female age, duration of subfertility, previous pregnancy, referral status, and semen analysis changed the probability to conceive spontaneously from 30% or greater to less than 30% in 97 of 3219 couples (3.0%). CONCLUSIONS: In ovulatory women, a basal FSH level of 8 IU/liter or higher is associated with decreasing fecundity, independent of female age and cycle length. Because the number of couples in whom the FSH level alters management decisions is low, we do not recommend routine testing of basal FSH in subfertile couples. PMID- 17405850 TI - Letrozole treatment of precocious puberty in girls with the McCune-Albright syndrome: a pilot study. AB - CONTEXT: Girls with McCune-Albright syndrome (MAS) and related disorders have gonadotropin-independent precocious puberty due to estrogen secretion from ovarian cysts. Their puberty does not respond to GnRH agonist therapy, and short acting aromatase inhibitors have had limited effectiveness. OBJECTIVE: Our objective was to assess the effectiveness of the potent, third-generation aromatase inhibitor letrozole in decreasing pubertal progression in girls with MAS and to assess the response of indices of bone turnover associated with the patients' polyostotic fibrous dysplasia. DESIGN: Subjects were evaluated at baseline and every 6 months for 12-36 months while on treatment with letrozole 1.5-2.0 mg/m(2).d. SETTING: This was an open-label therapeutic trial at a single clinical center. PATIENTS: Patients included nine girls aged 3-8 yr with MAS and/or gonadotropin-independent puberty. MAIN OUTCOME MEASURES: Measures included rates of linear growth, bone age advance, mean ovarian volume, estradiol, episodes of vaginal bleeding, and levels of the indices of bone metabolism: serum osteocalcin, alkaline phosphatase, urinary hydroxyproline, pyridinoline, deoxypyridinoline, and N-telopeptides. RESULTS: Girls had decreased rates of growth (P < or = 0.01) and bone age advance (P < or = 0.004) and cessation or slowing in their rates of bleeding over 12-36 months of therapy. Mean ovarian volume, estradiol, and indices of bone metabolism fell after 6 months (P < or = 0.05) but tended to rise by 24-36 months. Uterine volumes did not change. One girl had a ruptured ovarian cyst after 2 yr of treatment. CONCLUSIONS: This preliminary study suggests that letrozole may be effective therapy in some girls with MAS and/or gonadotropin-independent precocious puberty. Possible adverse effects include ovarian enlargement and cyst formation. PMID- 17405851 TI - Microglia activation in retinal degeneration. AB - Microglia cells are phagocytic sentinels in the CNS and in the retina required for neuronal homeostasis and innate immune defense. Accumulating experimental evidence suggests that chronic microglia activation is associated with various neurodegenerative diseases including retinal dystrophies. Endogenous triggers alert microglia cells rapidly in the degenerating retina, leading to local proliferation, migration, enhanced phagocytosis, and secretion of cytokines, chemokines, and neurotoxins. This amplified, immunological cascade and the loss of limiting control mechanisms may contribute significantly to retinal tissue damage and proapoptotic events. This review summarizes the developmental and immune surveillance functions of microglia in the healthy retina and discusses early signaling events and transcriptional networks of microglia activation in retinal degeneration. The characterization of activation pathways at the molecular level may lead to innovative, therapeutic options in degenerative retinal diseases based on a selective, pharmacological interference with the neurotoxic activities of microglia cells, without compromising their homeostastic functions. PMID- 17405852 TI - Adenosine receptor activation ameliorates type 1 diabetes. AB - Growing evidence indicates that adenosine receptors could be promising therapeutic targets in autoimmune diseases. Here we studied the role of adenosine receptors in controlling the course of type 1 diabetes. Diabetes in CD-1 mice was induced by multiple-low-dose-streptozotocin (MLDS) treatment and in nonobese diabetic (NOD) mice by cyclophosphamide injection. The nonselective adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA) prevented diabetes development in both MLDS-challenged mice and in cyclophosphamide-treated NOD mice. The effect of NECA was reversed by the selective A2B receptor antagonist N (4-cyanophenyl)-2-[4-(2,3,6,7-tetrahydro-2,6-dioxo-1,3-dipropyl-1H-purin-8 yl)phenoxy]acetamide (MRS 1754). The selective A1 receptor agonist 2-chloro-N6 cyclopentyladenosine (CCPA) and A3 receptor agonist N6-(3-iodobenzyl)-adenosine 5'-N-methyluronamide (IB-MECA) were less efficacious in ameliorating the course of diabetes. NECA inhibited diabetes in A2A receptor KO mice and the selective A2A receptor agonist 2-p-(2-carboxyethyl)phenethyl-amino-5'-N-ethyl carboxamidoadenosine (CGS21680) had no effect in normal mice, indicating a lack of role of A2A receptors. NECA failed to prevent cytokine-induced beta-cell death in vitro, but NECA strongly suppressed expression of the proinflammatory cytokines TNF-alpha, MIP-1alpha, IL-12, and IFN-gamma in pancreata, endotoxin, or anti-CD3-stimulated splenic cells, and T helper 1 lymphocytes, indicating that the beneficial effect of NECA was due to immunomodulation. These results demonstrate that adenosine receptor ligands are potential candidates for the treatment of type 1 diabetes. PMID- 17405853 TI - Exocytosis of norepinephrine at axon varicosities and neuronal cell bodies in the rat brain. AB - Norepinephrine secretion from central neurons was widely assumed to occur by exocytosis, but the essential characteristics of this process remained unknown. We developed an approach to study it directly by amperometry using carbon fiber microelectrodes in organotypic rat brainstem slice cultures. Noradrenergic neurons from areas A1 and A2 were fluorescently labeled by an adenoviral vector with noradrenergic-specific promoter. Quantal events, consistent with exocytotic release of norepinephrine, were registered at noradrenergic axonal varicosities as well as at cell bodies. According to their charge integrals, events were grouped into two populations. The majority (approximately 40 fC) were compatible with full exocytotic fusion of small clear and dense core vesicles shown in previous morphometric studies. The quantal size distribution was modulated by treatment with reserpine and amitriptyline. In addition, much larger quantal events (>1 pC) occurred at predominantly axonal release sites. The time course of signals was severalfold faster than in adrenal chromaffin cells, suggesting profound differences in the release machinery between these cell types. Tetrodotoxin eliminated the majority of events, indicating that release was partially, but not entirely, action potential driven. In conclusion, central norepinephrine release has unique characteristics, distinguishing it from those of other monoaminergic cells in periphery and brain. PMID- 17405854 TI - Clinical trials: the viewpoint of children. AB - The views of 30 children (8-16 years old) attending paediatric medical clinics on paediatric clinical trials were determined by semi-structured interviews. Nineteen children recognised that there were risks involved with taking part in clinical trials. Risks concerned with being paid were recognised by all children. PMID- 17405855 TI - The continuing decline in SIDS mortality. AB - The "Back to Sleep" campaign resulted in a dramatic decrease in sudden infant death syndrome (SIDS) worldwide. SIDS mortality has continued to decline (in New Zealand by 63% from 1993 to 2004), but the reason for this has not been explained. A postal survey found that the proportion of infants sleeping on their back has increased substantially (from 24.4% in 1992 to 72.3% in 2005), and this could account for the 39%-48% decrease in SIDS mortality. PMID- 17405856 TI - Increasing prevalence of hypospadias in Western Australia, 1980-2000. AB - OBJECTIVES: Hypospadias, a common birth defect, has shown widespread variation in reported rates and temporal trends across countries over the last 30 years. The aim of this study was to determine the prevalence and trends of hypospadias in an Australian population. DESIGN: Population-based study of all male infants born in Western Australia (WA) between 1980 and 2000 diagnosed with hypospadias and notified to the WA Birth Defects Registry. MAIN OUTCOME MEASURES: Prevalence of hypospadias, birth outcome and association with other congenital anomalies, stratified by degree-of-severity. RESULTS: 1788 cases of hypospadias were registered in WA in 1980-2000 with an overall prevalence of 34.8 (95% confidence interval (CI): 33.2 to 36.4) cases per 10 000 births. The prevalence increased by 2.0% per annum (95% CI: 1.2% to 2.8%) from 27.9 in 1980 to 43.2 per 10 000 births in 2000 (p<0.001). Hypospadias was mild in 84% of cases, moderate-severe in 11% and unspecified in 5%, with the number of moderate-severe hypospadias almost doubling over time (p<0.01). There were 1465 (82%) cases of isolated hypospadias and 323 (18%) had co-existing anomalies. Infants with co-existing genital (relative risk (RR) 4.5; 95% CI: 3.3 to 6.1) or non-genital (RR 1.5; 95% CI: 1.0 to 2.2) anomalies were more likely to have moderate-severe hypospadias compared with isolated cases. CONCLUSION: Hypospadias affects one in 231 births and has been reported to have increased significantly over the last 20 years. Future investigation of the aetiology of hypospadias is important to identify potentially modifiable risk factors and ensure optimal male reproductive health in the future. PMID- 17405857 TI - Severe receptive language disorder in childhood--familial aspects and long-term outcomes: results from a Scottish study. AB - BACKGROUND AND AIMS: Little is known about the familial characteristics of children with severe receptive specific language impairment (SLI). Affected children are more likely to have long-term problems than those with expressive SLI but to date they have only been described as small cohorts within SLI populations. We therefore aimed to describe the clinical and familial characteristics of severe receptive SLI as defined by a rigorous phenotype and to establish whether non-word repetition showed a relationship with language impairment in these families. METHODS: Cross-sectional study of children who met ICD-10 (F80.2) criteria for receptive SLI at school entry, their siblings and genetic parents with standardised measures of language and non-verbal IQ, phonological auditory memory and speech sound inventory. RESULTS: At a mean of 6 years after school entry with a severe receptive SLI, the 58 participants had a normal mean and standard deviation non-verbal IQ, but only 3% (two) had attained language measures in the normal range. One third still had severe receptive language impairment. One third of siblings not known to be affected had language levels outside the normal range. Phonological auditory memory was impaired in most family members. CONCLUSION: Severe receptive SLI is nearly always associated with an equally severe reduction in expressive language skills. Language impairment in siblings may go undetected and yet they are at high risk. Family members had weak phonological auditory memory skills, suggesting that this could be a marker for language acquisition difficulties. Receptive SLI rarely resolves and trials of therapy are urgently needed. PMID- 17405858 TI - No temporal association between influenza outbreaks and invasive pneumococcal infections. AB - OBJECTIVE: To assess whether the influenza peak in populations precedes the annual peak for invasive pneumococcal infections (IPI) in winter. DESIGN: Ecological study. Active surveillance data on influenza A and IPI in children up to 16 years of age collected from 1997 to 2003 were analysed. SETTING: Paediatric hospitals in Germany. PATIENTS: Children under 16 years of age. RESULTS: In all years under study, the influenza A season did not appear to affect the IPI season (p = 0.49). Specifically, the influenza peak never preceded the IPI peak. CONCLUSION: On a population level there was no indication that the annual influenza epidemic triggered the winter increase in the IPI rate or the peak of the IPI distribution in children. PMID- 17405859 TI - Structural basis for ligand and heparin binding to neuropilin B domains. AB - Neuropilin (Nrp) is a cell surface receptor with essential roles in angiogenesis and axon guidance. Interactions between Nrp and the positively charged C termini of its ligands, VEGF and semaphorin, are mediated by Nrp domains b1 and b2, which share homology to coagulation factor domains. We report here the crystal structure of the tandem b1 and b2 domains of Nrp-1 (N1b1b2) and show that they form a single structural unit. Cocrystallization of N1b1b2 with Tuftsin, a peptide mimic of the VEGF C terminus, reveals the site of interaction with the basic tail of VEGF on the b1 domain. We also show that heparin promotes N1b1b2 dimerization and map the heparin binding site on N1b1b2. These results provide a detailed picture of interactions at the core of the Nrp signaling complex and establish a molecular basis for the synergistic effects of heparin on Nrp mediated signaling. PMID- 17405860 TI - ATM deficiency impairs thymocyte maturation because of defective resolution of T cell receptor alpha locus coding end breaks. AB - The ATM (ataxia telangiectasia mutated) protein plays a central role in sensing and responding to DNA double-strand breaks. Lymphoid cells are unique in undergoing physiologic double-strand breaks in the processes of Ig class switch recombination and T or B cell receptor V(D)J recombination, and a role for ATM in these processes has been suggested by clinical observations in ataxia telangiectasia patients as well as in engineered mice with mutations in the Atm gene. We demonstrate here a defect in thymocyte maturation in ATM-deficient mice that is associated with decreased efficiency in V-J rearrangement of the endogenous T cell receptor (TCR)alpha locus, accompanied by increased frequency of unresolved TCR Jalpha coding end breaks. We also demonstrate that a functionally rearranged TCRalphabeta transgene is sufficient to restore thymocyte maturation, whereas increased thymocyte survival by bcl-2 cannot improve TCRalpha recombination and T cell development. These data indicate a direct role for ATM in TCR gene recombination in vivo that is critical for surface TCR expression in CD4(+)CD8(+) cells and for efficient thymocyte selection. We propose a unified model for the two major clinical characteristics of ATM deficiency, defective T cell maturation and increased genomic instability, frequently affecting the TCRalpha locus. In the absence of ATM, delayed TCRalpha coding joint formation results both in a reduction of alphabeta TCR-expressing immature cells, leading to inefficient thymocyte selection, and in accumulation of unstable open chromosomal DNA breaks, predisposing to TCRalpha locus-associated chromosomal abnormalities. PMID- 17405861 TI - Structure and evolution of the Ivy protein family, unexpected lysozyme inhibitors in Gram-negative bacteria. AB - Part of an ancestral bactericidal system, vertebrate C-type lysozyme targets the peptidoglycan moiety of bacterial cell walls. We report the crystal structure of a protein inhibitor of C-type lysozyme, the Escherichia coli Ivy protein, alone and in complex with hen egg white lysozyme. Ivy exhibits a novel fold in which a protruding five-residue loop appears essential to its inhibitory effect. This feature guided the identification of Ivy orthologues in other Gram-negative bacteria. The structure of the evolutionary distant Pseudomonas aeruginosa Ivy orthologue was also determined in complex with hen egg white lysozyme, and its antilysozyme activity was confirmed. Ivy expression protects porous cell-wall E. coli mutants from the lytic effect of lysozyme, suggesting that it is a response against the permeabilizing effects of the innate vertebrate immune system. As such, Ivy acts as a virulence factor for a number of Gram-negative bacteria infecting vertebrates. PMID- 17405862 TI - On the path to extinction. AB - Populations can die out in many ways. We investigate one basic form of extinction, stable or intrinsic extinction, caused by individuals on the average not being able to replace themselves through reproduction. The archetypical such population is a subcritical branching process, i.e., a population of independent, asexually reproducing individuals, for which the expected number of progeny per individual is less than one. The main purpose is to uncover a fundamental pattern of nature. Mathematically, this emerges in large systems, in our case subcritical populations, starting from a large number, x, of individuals. First we describe the behavior of the time to extinction T: as x grows to infinity, it behaves like the logarithm of x, divided by r, where r is the absolute value of the Malthusian parameter. We give a more precise description in terms of extreme value distributions. Then we study population size partway (or u-way) to extinction, i.e., at times uT, for 0 < u < 1, e.g., u = 1/2 gives halfway to extinction. (Note that mathematically this is no stopping time.) If the population starts from x individuals, then for large x, the proper scaling for the population size at time uT is x into the power u - 1. Normed by this factor, the population u-way to extinction approaches a process, which involves constants that are determined by life span and reproduction distributions, and a random variable that follows the classical Gumbel distribution in the continuous time case. In the Markov case, where an explicit representation can be deduced, we also find a description of the behavior immediately before extinction. PMID- 17405863 TI - Microarray analysis of newly synthesized RNA in cells and animals. AB - Current methods to analyze gene expression measure steady-state levels of mRNA. To specifically analyze mRNA transcription, we have developed a technique that can be applied in vivo in intact cells and animals. Our method makes use of the cellular pyrimidine salvage pathway and is based on affinity-chromatographic isolation of thiolated mRNA. When combined with data on mRNA steady-state levels, this method is able to assess the relative contributions of mRNA synthesis and degradation/stabilization. It overcomes limitations associated with currently available methods such as mechanistic intervention that disrupts cellular physiology, or the inability to apply the techniques in vivo. Our method was first tested in serum response of cultured fibroblast cells and then applied to the study of renal ischemia reperfusion injury, demonstrating its applicability for whole organs in vivo. Combined with data on mRNA steady-state levels, this method provided a detailed analysis of regulatory mechanisms of mRNA expression and the relative contributions of RNA synthesis and turnover within distinct pathways, and identification of genes expressed at low abundance at the transcriptional level. PMID- 17405864 TI - Extracellular RNA constitutes a natural procoagulant cofactor in blood coagulation. AB - Upon vascular injury, locally controlled haemostasis prevents life-threatening blood loss and ensures wound healing. Intracellular material derived from damaged cells at these sites will become exposed to blood components and could contribute to blood coagulation and pathological thrombus formation. So far, the functional and mechanistic consequences of this concept are not understood. Here, we present in vivo and in vitro evidence that different forms of eukaryotic and prokaryotic RNA serve as promoters of blood coagulation. Extracellular RNA was found to augment (auto-)activation of proteases of the contact phase pathway of blood coagulation such as factors XII and XI, both exhibiting strong RNA binding. Moreover, administration of exogenous RNA provoked a significant procoagulant response in rabbits. In mice that underwent an arterial thrombosis model, extracellular RNA was found associated with fibrin-rich thrombi, and pretreatment with RNase (but not DNase) significantly delayed occlusive thrombus formation. Thus, extracellular RNA derived from damaged or necrotic cells particularly under pathological conditions or severe tissue damage represents the long sought natural "foreign surface" and provides a procoagulant cofactor template for the factors XII/XI-induced contact activation/amplification of blood coagulation. Extracellular RNA thereby reveals a yet unrecognized target for antithrombotic intervention, using RNase or related therapeutic strategies. PMID- 17405865 TI - Complete genetic linkage can subvert natural selection. AB - The intricate adjustment of organisms to their environment demonstrates the effectiveness of natural selection. But Darwin himself recognized that certain biological features could limit this effectiveness, features that generally reduce the efficiency of natural selection or yield suboptimal adaptation. Genetic linkage is known to be one such feature, and here we show theoretically that it can introduce a more sinister flaw: when there is complete linkage between loci affecting fitness and loci affecting mutation rate, positive natural selection and recurrent mutation can drive mutation rates in an adapting population to intolerable levels. We discuss potential implications of this finding for the early establishment of recombination, the evolutionary fate of asexual populations, and immunological clearance of clonal pathogens. PMID- 17405866 TI - Central nervous system drug disposition: the relationship between in situ brain permeability and brain free fraction. AB - The dispositions of 50 marketed central nervous system (CNS) drugs into the brain have been examined in terms of their rat in situ (P) and in vitro apparent membrane permeability (P(app)) alongside lipophilicity and free fraction in rat brain tissue. The inter-relationship between these parameters highlights that both permeability and brain tissue binding influence the uptake of drugs into the CNS. Hydrophilic compounds characterized by low brain tissue binding display a strong correlation (R(2) = 0.82) between P and P(app), whereas the uptake of more lipophilic compounds seems to be influenced by both P(app) and brain free fraction. A nonlinear relationship is observed between logP(oct) and P over the 6 orders of magnitude range in lipophilicity studied. These findings corroborate recent reports in the literature that brain penetration is a function of both rate and extent of drug uptake into the CNS. PMID- 17405867 TI - Pharmacological effects of ephedrine alkaloids on human alpha(1)- and alpha(2) adrenergic receptor subtypes. AB - Ephedra species of plants have both beneficial and adverse effects primarily associated with the presence of ephedrine alkaloids. Few reports have appeared that examine the direct actions of ephedrine alkaloids on human subtypes of adrenergic receptors (ARs). In the present study, ephedrine alkaloids were evaluated for their binding affinities on human alpha(1A)-, alpha(1B)-, alpha(1D) , alpha(2A)-, alpha(2B)-, and alpha(2C)-AR subtypes expressed in HEK and Chinese hamster ovary cells. Cell-based reporter gene assays were used to establish functional activity of ephedrine alkaloids at alpha(1A)-, alpha(2A)-, and alpha(2C)-ARs. The data showed that ephedrine alkaloids did not activate alpha(1) and alpha(2)-ARs and that they antagonized the agonist-mediated effects of phenylephrine and medetomidine on alpha(1)- and alpha(2)-ARs, respectively. As in the binding studies, 1R,2R- and 1R,2S-ephedrine showed greater functional antagonist activity than the 1S,2R- and 1S,2S-isomers. The rank order of affinity for the isomers was 1R,2R > 1R,2S > 1S,2R > 1S,2S. The rank order of potencies of alkaloids containing a 1R,2S-configuration was norephedrine > or = ephedrine >> N methylephedrine. These studies have demonstrated that orientation of the beta hydroxyl group on the ethylamino side chain and the state of N-methyl substitution are important for alpha-AR binding and functional activity of the ephedrine alkaloids. In conclusion, the ephedrine isomers and analogs studied did not exhibit any direct agonist activity and were found to possess moderate antagonist activities on cloned human alpha-ARs. The blockade of presynaptic alpha(2A)- and alpha(2C)-ARs may have a pharmacological role in the direct actions of Ephedra alkaloids. PMID- 17405868 TI - Flufenamic acid bi-directionally modulates the transient outward K(+) current in rat cerebellar granule cells. AB - In this report, the effect of flufenamic acid on voltage-activated transient outward K(+) current (I(A)) in cultured rat cerebellar granule cells was investigated. At a concentration of 20 microM to 1 mM, flufenamic acid reversibly inhibited I(A) in a dose-dependent manner. However, flufenamic acid at a concentration of 0.1 to 10 microM significantly increased the current amplitude of I(A). In addition to the current amplitude of I(A), a higher concentration of flufenamic acid had a significant effect on the kinetic parameters of the steady state activation and inactivation process, suggesting that the binding affinity of flufenamic acid to I(A) channels may be state-dependent. Silencing the K(v)4.2, K(v)4.3, and K(v)1.1 genes of I(A) channels using small interfering RNA did not change the inhibitory effect of flufenamic on I(A), indicating that flufenamic acid did not act specifically on any of the subunits of the I(A) channel protein. Intracellular application of flufenamic acid could significantly increase the I(A) amplitude but did not alter the inhibited effect induced by extracellular application of flufenamic acid, implying that flufenamic acid may exert its effect from both the inside and outside sites of the channel. Furthermore, the activation of current induced by intracellular application of flufenamic acid could mimic other cyclooxygenase inhibitors and arachidonic acid. Our data are the first that demonstrate how flufenamic acid is able to bidirectionally modulate I(A) channels in neurons at different concentrations and by different methods of application and that two different mechanisms may be involved. PMID- 17405869 TI - Decrease in N-methyl-D-aspartic acid receptor-NR2B subunit levels by intrathecal short-hairpin RNA blocks group I metabotropic glutamate receptor-mediated hyperalgesia. AB - The present study characterizes the involvement of the N-methyl-D-aspartic acid receptors (NMDARs) in mediating thermal hyperalgesia induced by activation of group I metabotropic glutamate receptors (mGluRs). Intrathecal administration of the mGluR1/5 agonist (S)-3,5-DHPG [(S)-3,5-dihydroxyphenylglycine] to mice resulted in significant hyperalgesia as assessed by the tail immersion test. The pretreatment of mice i.t. with CGS 19755 (selective antagonist of the NMDAR), CGP 78608 [[(1S)-1-[[(7-bromo-1,2,3,4-tetrahydro-2,3-dioxo-5 quinoxalinyl)methyl]amino]ethyl]phosphonic acid] (selective antagonist at the glycine-binding site of the NMDAR), ifenprodil and Ro 25-6981 (selective antagonists of the NR2B subunit of the NMDAR), bisindolylmaleimide I and Go-7874 [12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo(2,3 a)pyrrolo(3,4-c)-carbazole] (inhibitors of protein kinase C), or PKI-(14-22) amide [Myr-N-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile-NH(2)] (inhibitor of protein kinase A) dose-dependently inhibited the hyperalgesia induced by i.t. administration of the mGluR1/5 receptor agonist (S)-3,5-DHPG. In contrast, i.t. pretreatment of mice with NVP-AAM077 [[(R)-[(S)-1-(4-bromophenyl)-ethylamino] (2,3-dioxo-1,2,3,4-tetrahydroquinoxalin-5-yl)-methyl]-phosphonic acid] (selective antagonist of the NR2A subunit of the NMDAR) or DT-3 [H-Arg-Gln-Ile-Lys-Ile-Trp Phe-Gln-Asn-Arg-Arg-Met-Lys-Trp-Lys-Lys-Leu-Arg-Lys-Lys-Lys-Lys-Lys-His-OH] (inhibitor of protein kinase G) had no effect on (S)-3,5-DHPG-mediated hyperalgesia. We also show for the first time that i.t. injection of pSM2 (pShag Magic version 2)-grin2b (coding for an short-hairpin RNA to the NR2B subunit of the NMDAR) resulted in a dose-dependent decrease in the NR2B protein and blockade of hyperalgesia induced by activation of the mGluR1/5 in (S)-3,5-DHPG-treated mice. Taken together, our results suggest the hypothesis that mGluRs are coupled to the NMDAR channels through the NR2B subunit in the spinal cord and that this coupling involves the activation of protein kinase C and protein kinase A. PMID- 17405870 TI - Volume guarantee versus high-frequency ventilation: lung inflammation in preterm infants. AB - BACKGROUND: Appropriate ventilation together with improvement of clinical care of premature babies can contribute to reducing lung inflammation, known to represent the "primum movens" of bronchopulmonary dysplasia (BPD). High-frequency oscillatory ventilation (HFOV) and volume-guarantee (VG) ventilation are effective in the treatment of neonatal respiratory distress syndrome (RDS). OBJECTIVE: To assess the potential of HFOV and VG to prevent BPD in the acute phase of RDS, by a randomised clinical study evaluating lung inflammation in premature infants. STUDY DESIGN: Forty infants (gestational age 25-32 weeks) with RDS were assigned to assist-control ventilation plus VG (Vt = 5 ml/kg) or HFOV (both with a Drager Babylog 8000 plus ventilator). Levels of interleukin (IL) 6, IL8 and tumour necrosis factor were determined in tracheal aspirate on days 1, 3 and 7 of life. RESULTS: In the HFOV group IL6 levels were significantly higher on day 3 (0.5 (0.2) vs assisted-control ventilation plus VG group 0.1 (0.2) ng/ml) and oxygen dependency was significantly longer (36 (23) vs assisted-control ventilation plus VG group 19 (11) days). CONCLUSION: VG ventilation is an effective lung-protective strategy to be used in acute RDS, inducing a lower expression of early inflammation markers than HFOV. Whether the use of this initial ventilatory strategy contributes to the prevention of BPD requires further studies. PMID- 17405871 TI - Fetal cortisol response to intrauterine microbial colonisation identified by the polymerase chain reaction and fetal inflammation. AB - OBJECTIVE: To determine the fetal cortisol response to intrauterine infection. STUDY DESIGN: 16s ribosomal RNA genes or the urease genes of Ureaplasma spp were identified by the polymerase chain reaction in intrauterine samples. Cord blood cortisol, interleukin 6 (IL6) and IL8 were measured in samples from 31 infants born at <32 weeks gestation. RESULTS: 13 infants (median gestation 30 weeks, birth weight 1350 g) had at least one positive intrauterine sample for microbial genes and 18 infants (31 weeks, 1320 g) did not. The cord blood cortisol concentration was significantly higher in fetuses exposed to intrauterine infection and significantly increased in fetuses/mothers presenting in preterm labour with intact membranes compared with infants delivered by elective prelabour caesarean section (p<0.05). The cord blood cortisol concentration was increased in the mothers with prelabour premature rupture of membranes but this was not significant compared with the caesarean section group. The cord blood cortisol concentration was significantly increased in the presence of chorioamnionitis or funisitis and was moderately correlated with cord blood IL6 (r = 0.64, p<0.01) and IL8 (r = 0.52, p<0.01). CONCLUSIONS: In this study, cord blood cortisol was increased in the colonised group compared with non-colonised infants. It is unclear if infants born following prelabour premature rupture of the membranes mount an adequate anti-inflammatory response. PMID- 17405872 TI - Building a pharmacological lexicon: small molecule discovery in academia. AB - Small molecules are powerful pharmacological tools to dissect biological events. There is now considerable interest in expanding efforts to identify and use small molecules targeting proteins encoded in the genomes of humans and pathogenic organisms. Integration of the principles of molecular pharmacology with contemporary high-throughput and high-content screening technologies is essential for the success of these discovery activities. We present some of the challenges and opportunities provided by the Molecular Library Screening Centers Network (MLSCN), which is a National Institutes of Health Roadmap Initiative. PMID- 17405873 TI - Crystal structure of the cysteine-rich domain of scavenger receptor MARCO reveals the presence of a basic and an acidic cluster that both contribute to ligand recognition. AB - MARCO is a trimeric class A scavenger receptor of macrophages and dendritic cells that recognizes polyanionic particles and pathogens. The distal, scavenger receptor cysteine-rich (SRCR) domain of the extracellular part of this receptor has been implicated in ligand binding. To provide a structural basis for understanding the ligand-binding mechanisms of MARCO, we have determined the crystal structure of the mouse MARCO SRCR domain. The recombinant SRCR domain purified as monomeric and dimeric forms, and their structures were determined at 1.78 and 1.77 A resolution, respectively. The monomer has a compact globular fold with a twisted five-stranded antiparallel beta-sheet and a long loop covering a single alpha-helix, whereas the dimer is formed via beta-strand swapping of two monomers, thus containing a large eight-stranded beta-sheet. Calculation of the surface electrostatic potential revealed that the beta-sheet region with several arginines forms a basic cluster. Unexpectedly, an acidic cluster was found in the long loop region. In the monomer, the acidic cluster is involved in metal ion binding. Studies with cells expressing various SRCR domain mutants showed that all of the arginines of the basic cluster are involved in ligand binding, suggesting a cooperative binding mechanism. Ligand binding is also dependent on the acidic cluster and Ca2+ ions whose depletion appears to affect ligand binding at least by modulating the electrostatic potential or relative domain orientation. We propose that the SRCR domain dimerization can contribute to the recognition of large ligands by providing a means for the MARCO receptor oligomerization. PMID- 17405874 TI - Peroxisomal proliferator-activated receptor-alpha-dependent inhibition of endothelial cell proliferation and tumorigenesis. AB - The peroxisomal proliferator-activated nuclear receptor-alpha (PPARalpha), the target for most hypolipidemic drugs in current clinical use, regulates the transcription of genes involved in lipid metabolism and transport, and energy homeostasis. More recently, PPARalpha and its ligands have been implicated in inflammatory responses and the regulation of cell proliferation. PPARalpha also regulates the expression of Cyp4a fatty acid omega-hydroxylases and Cyp2c arachidonic acid epoxygenase genes. To study the role of the PPARalpha receptor and of its Cyp2c epoxygenase gene target in tumorigenesis, we treated mice injected with tumor cells with Wy-14,643, a PPARalpha-selective ligand. Compared with untreated controls, Wy-14643-treated animals showed marked reductions in tumor growth and vascularization, which were accompanied by decreases in the plasma levels of pro-angiogenic epoxygenase metabolites (EETs), hepatic EET biosynthesis, and Cyp2c epoxygenase expression. All these Wy-14643-induced responses were absent in PPARalpha(-/-) mice and are thus PPARalpha-mediated. Primary cultures of mouse lung endothelial cells treated with Wy-14643 showed reductions in cell proliferation and in the formation of capillary-like structures. These effects were absent in cells obtained from PPRAalpha(-/-) mice and reversed by the addition of EETs. These results identify important anti angiogenic and anti-tumorigenic roles for PPARalpha, characterize the contribution of its Cyp2c epoxygenases gene target to these responses, and suggest potential anti-cancer roles for this nuclear receptor and its ligands. PMID- 17405875 TI - Exoribonuclease R in Pseudomonas syringae is essential for growth at low temperature and plays a novel role in the 3' end processing of 16 and 5 S ribosomal RNA. AB - The (3'-->5') exoribonuclease RNase R interacts with the endoribonuclease RNase E in the degradosome of the cold-adapted bacterium Pseudomonas syringae Lz4W. We now present evidence that the RNase R is essential for growth of the organism at low temperature (4 degrees C). Mutants of P. syringae with inactivated rnr gene (encoding RNase R) are cold-sensitive and die upon incubation at 4 degrees C, a phenotype that can be complemented by expressing RNase R in trans. Overexpressing polyribonucleotide phosphorylase in the rnr mutant does not rescue the cold sensitivity. This is different from the situation in Escherichia coli, where rnr mutants show normal growth, but pnp (encoding polyribonucleotide phosphorylase) and rnr double mutants are nonviable. Interestingly, RNase R is not cold inducible in P. syringae. Remarkably, however, rnr mutants of P. syringae at low temperature (4 degrees C) accumulate 16 and 5 S ribosomal RNA (rRNA) that contain untrimmed extra ribonucleotide residues at the 3' ends. This suggests a novel role for RNase R in the rRNA 3' end processing. Unprocessed 16 S rRNA accumulates in the polysome population, which correlates with the inefficient protein synthesis ability of mutant. An additional role of RNase R in the turnover of transfer-messenger RNA was identified from our observation that the rnr mutant accumulates transfer-messenger RNA fragments in the bacterium at 4 degrees C. Taken together our results establish that the processive RNase R is crucial for RNA metabolism at low temperature in the cold-adapted Antarctic P. syringae. PMID- 17405876 TI - Inhibiting proteasomal degradation of microsomal triglyceride transfer protein prevents CCl4-induced steatosis. AB - Carbon tetrachloride (CCl(4)) interferes with triglyceride secretion and causes steatosis, fibrosis, and necrosis. In mice, CCl(4) decreased plasma triglyceride rich lipoproteins, increased cellular lipids, and reduced microsomal triglyceride transfer protein (MTP) without diminishing mRNA levels. Similarly, CCl(4) decreased apoB-lipoprotein production and MTP activity but had no effect on mRNA levels in primary enterocytes and colon carcinoma and hepatoma cells. CCl(4) did not affect MTP synthesis but induced post-translational degradation involving ubiquitinylation and proteasomes in McA-RH7777 cells. By contrast, MTP inhibitor increased cellular lipids without affecting MTP protein. MTP was covalently modified when cells were incubated with (14)CCl(4). This modification was prevented by the inhibition of P450 oxygenases, indicating that CCl(3)(.) generated by these enzymes targets MTP for degradation. To determine whether inhibition of proteolysis could prevent CCl(4) toxicity, mice were fed with CCl(4) with or without lactacystin. Lactacystin increased ubiquitinylated MTP and prevented lipid accumulation in tissues. Thus, CCl(4) induces post-translational degradation without affecting lipid transfer activity, whereas MTP antagonist inhibits lipid transfer activity without causing its destruction. These studies identify MTP as a major target of CCl(4) and its degradation as a novel mechanism involved in the onset of steatosis, suggesting that inhibition of proteolysis may prevent some forms of steatosis. PMID- 17405877 TI - Mechanisms for picrotoxinin and picrotin blocks of alpha2 homomeric glycine receptors. AB - Contrary to its effect on the gamma-aminobutyric acid type A and C receptors, picrotoxin antagonism of the alpha1 homomeric glycine receptors (GlyRs) has been shown to be non-use-dependent and nonselective between the picrotoxin components picrotoxinin and picrotin. Picrotoxin antagonism of the embryonic alpha2 homomeric GlyR is known to be use-dependent and reflects a channel-blocking mechanism, but the selectivity of picrotoxin antagonism of the embryonic alpha2 homomeric GlyRs between picrotoxinin and picrotin is unknown. Hence, we used the patch clamp recording technique in the outside-out configuration to investigate, at the single channel level, the mechanism of picrotin- and picrotoxinin-induced inhibition of currents, which were evoked by the activation of alpha2 homomeric GlyRs stably transfected into Chinese hamster ovary cells. Although both picrotoxinin and picrotin inhibited glycine-evoked outside-out currents, picrotin had a 30 times higher IC50 than picrotoxinin. Picrotin-evoked inhibition displayed voltage dependence, whereas picrotoxinin did not. Picrotoxinin and picrotin decreased the mean open time of the channel in a concentration-dependent manner, indicating that these picrotoxin components can bind to the receptor in its open state. When picrotin and glycine were co-applied, a large rebound current was observed at the end of the application. This rebound current was considerably smaller when picrotoxinin and glycine were co-applied. Both picrotin and picrotoxinin were unable to bind to the unbound conformation of the receptor, but both could be trapped at their binding site when the channel closed during glycine dissociation. Our data indicate that picrotoxinin and picrotin are not equivalent in blocking alpha2 homomeric GlyR. PMID- 17405878 TI - Crystal structure of human cytosolic 5'-nucleotidase II: insights into allosteric regulation and substrate recognition. AB - Cytosolic 5'-nucleotidase II catalyzes the dephosphorylation of 6-hydroxypurine nucleoside 5'-monophosphates and regulates the IMP and GMP pools within the cell. It possesses phosphotransferase activity and thereby also catalyzes the reverse reaction. Both reactions are allosterically activated by adenine-based nucleotides and 2,3-bisphosphoglycerate. We have solved structures of cytosolic 5'-nucleotidase II as native protein (2.2 Angstrom) and in complex with adenosine (1.5 Angstrom) and beryllium trifluoride (2.15 Angstrom) The tetrameric enzyme is structurally similar to enzymes of the haloacid dehalogenase (HAD) superfamily, including mitochondrial 5'(3')-deoxyribonucleotidase and cytosolic 5' nucleotidase III but possesses additional regulatory regions that contain two allosteric effector sites. At effector site 1 located near a subunit interface we modeled diadenosine tetraphosphate with one adenosine moiety in each subunit. This efficiently glues the tetramer subunits together in pairs. The model shows why diadenosine tetraphosphate but not diadenosine triphosphate activates the enzyme and supports a role for cN-II during apoptosis when the level of diadenosine tetraphosphate increases. We have also modeled 2,3 bisphosphoglycerate in effector site 1 using one phosphate site from each subunit. By comparing the structure of cytosolic 5'-nucleotidase II with that of mitochondrial 5'(3')-deoxyribonucleotidase in complex with dGMP, we identified residues involved in substrate recognition. PMID- 17405879 TI - Regulation of copper homeostasis by micro-RNA in Arabidopsis. AB - Major copper proteins in the cytoplasm of plant cells are plastocyanin, copper/zinc superoxide dismutase, and cytochrome c oxidase. Under copper limited conditions, expression of copper/zinc superoxide dismutase is down-regulated and the protein is replaced by iron superoxide dismutase in chloroplasts. We present evidence that a micro-RNA, miR398, mediates this regulation in Arabidopsis thaliana, by directing the degradation of copper/zinc superoxide dismutase mRNA when copper is limited. Sequence analysis indicated that the transcripts encoding cytosolic copper/zinc superoxide dismutase and COX5b-1, a subunit of the mitochondrial cytochrome c oxidase, are also targeted by miR398. This regulation via miR398 takes place in response to changes in a low range of copper levels (0.2-0.5 microM), indicating that miR398 is involved in a response to copper limitation. On the other hand, another major copper protein, plastocyanin, which is involved in photosynthetic electron flow and is essential in higher plants, was not regulated via miR398. We propose that miR398 is a key factor in copper homeostasis in plants and regulates the stability of mRNAs of major copper proteins under copper-limited conditions. PMID- 17405880 TI - Closed-state cross-linking of adjacent beta1 subunits in alpha1beta1 GABAa receptors via introduced 6' cysteines. AB - The pore structural changes associated with Cys-loop receptor gating are currently the subject of considerable interest. Several functional approaches have shown that surface exposure of pore-lining side chains does not change significantly during activation. However, a disulfide trapping study on alpha1(T6'C)beta1(T6'C) gamma-aminobutyric acid type A (GABA(A)) receptors (GABA(A)Rs), which showed that adjacent beta subunits cross-link in the open state only, concluded that channel gating is mediated by beta subunits contra rotating through a summed angle of approximately 120 degrees. Such a large rotation is not easily reconciled with other evidence. The present study initially sought to investigate an observation that appeared inconsistent with the rotation model: namely that alpha1(T6'C)beta1(T6'C) GABA(A)Rs expressed in HEK293 cells form 6' cysteine-mediated disulfide bonds in the closed state. On the basis of electrophysiological and Western blotting experiments, we conclude that adjacent beta(T6'C) subunits dimerise efficiently in the closed state via cross-links between their respective 6' cysteines and that this locks the channels closed. This questions the beta subunit contra-rotation model of activation and raises the question of how the closed state cross-links form. We propose that beta subunit 6' cysteines move into sufficiently close proximity for disulfide formation via relatively large amplitude random thermal motions that appear to be a unique feature of beta subunits. Because dimerized channels are locked closed, we conclude either that the spontaneous beta subunit movements or asymmetries in the movements of adjacent beta subunits during activation are essential for pore opening. Our results identify a novel feature of GABA(A)R gating that may be important for understanding its activation mechanism. PMID- 17405881 TI - Intersectin-1s regulates the mitochondrial apoptotic pathway in endothelial cells. AB - Intersectins (ITSNs) are multidomain adaptor proteins implicated in endocytosis, regulation of actin polymerization, and Ras/MAPK signaling. We have previously shown that ITSN-1s is required for caveolae fission and internalization in endothelial cells (ECs). In the present study, using small interfering RNA to knock down ITSN-1s protein expression, we demonstrate a novel role of ITSN-1s as a key antiapoptotic protein. Knockdown of ITSN-1s in ECs activated the mitochondrial pathway of apoptosis as determined by genomic DNA fragmentation, extensive mitochondrial fission, activation of the proapoptotic proteins BAK and BAX, and cytochrome c efflux from mitochondria. ITSN-1 knockdown acts as a proapoptotic signal that causes mitochondrial outer membrane permeabilization, dissipation of the mitochondrial membrane potential, and generation of reactive oxygen species. These effects were secondary to decreased activation of Erk1/2 and its direct activator MEK. Bcl-X(L) overexpression prevented BAX activation and the apoptotic ECs death induced by suppression of ITSN-1s. Our findings demonstrate a novel role of ITSN-1s as a negative regulator of the mitochondrial pathway-dependent apoptosis secondary to activation of the Erk1/2 survival signaling pathway. PMID- 17405882 TI - Mice deficient in heparan sulfate 6-O-sulfotransferase-1 exhibit defective heparan sulfate biosynthesis, abnormal placentation, and late embryonic lethality. AB - Heparan sulfate (HS) plays critical roles in a variety of developmental, physiological, and pathogenic processes due to its ability to interact in a structure-dependent manner with numerous growth factors that participate in cellular signaling. The divergent structures of HS glycosaminoglycans are the result of the coordinate actions of several N- and O-sulfotransferases, C5 epimerase, and 6-O-endosulfatases. We have shown that 6-O-sulfation of the glucosamine residues in HS are catalyzed by the sulfotransferases HS6ST-1, -2, and -3. To determine the biological and physiological importance of HS6ST-1, we now describe the creation of transgenic mice that lack this sulfotransferase. Most of our HS6ST-1-null mice died between embryonic day 15.5 and the perinatal stage, and those mice that survived were considerably smaller than their wild type littermates. Some of these HS6ST-1-null mice exhibited development abnormalities, and histochemical and molecular analyses of these mice revealed an approximately 50% reduction in the number of fetal microvessels in the labyrinthine zone of the placenta relative to that in the wild-type mice. Because we observed a modest reduction in VEGF-A mRNA and protein in the tissues of HS6ST 1-null mice, an HS-dependent defect in cytokine signaling probably contributes to increased embryonic lethality and decreased growth. Biochemical studies of the HS chains isolated from various organs of our HS6ST-1-null mice revealed a marked reduction of GlcNAc(6SO(4)) and HexA-GlcNSO(3)(6SO(4)) levels and a reduced ability to bind Wnt2. Thus, despite the presence of three closely related 6-O sulfotransferase genes in the mouse genome, HS6ST-1 is the primary one used in HS biosynthesis in most tissues. PMID- 17405883 TI - Influence of combined afferent stimulation and task-specific training following stroke: a pilot randomized controlled trial. AB - BACKGROUND: Reorganization of the human motor cortex can be induced by specific patterns of peripheral afferent stimulation. The potential for afferent stimulation to facilitate the functional recovery associated with conventional rehabilitative techniques has not previously been investigated. OBJECTIVE: The authors sought to determine whether combining appropriate afferent stimulation with task-specific training resulted in greater improvements than training alone in patients with impaired upper limb function in the subacute phase following stroke. METHOD: Twenty patients with hemiparesis due to stroke were allocated randomly to either a stimulation or control group. All received 9 sessions of task-specific physiotherapy training over 3 weeks. Prior to each training session, associative electrical stimulation of the motor point of 2 hand muscles was given in the stimulation group, whereas the control group received sham stimulation. Changes in dexterity were assessed using a grip-lift task, and standard measures of upper-limb function were made before and following the intervention. Corticospinal excitability was examined using transcranial magnetic stimulation. RESULTS: Both groups showed comparable improvements in functional measures of upper-limb function. Of the 20 patients, only 14 could perform the grip-lift task, which is an objective measure of dexterity. Patients in the stimulation group exhibited significantly greater improvements in this task than the control group. There was no significant change in corticospinal excitability in either group. CONCLUSION: This pilot study provides preliminary data suggesting that targeted afferent stimulation may facilitate the response to conventional rehabilitation in patients with hemiparesis due to stroke, but these results need to be confirmed in a larger scale study. PMID- 17405884 TI - Importance of sports in cartilage regeneration after autologous chondrocyte implantation: a prospective study with a 3-year follow-up. AB - BACKGROUND: There have been no data in the literature reporting the influence of sports on the outcome of autologous chondrocyte implantation in chondral defects of the knee. HYPOTHESIS: Sports can improve the result of autologous chondrocyte implantation in postoperative follow-up. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Between 1997 and 2003, 118 patients with symptomatic isolated chondral lesions of the knee underwent autologous chondrocyte implantation. According to the sports activity level before the start of symptoms, patients were assigned to 2 groups: group II with no or rare sports involvement (1-3 times/month); group I with regular (1-3 times/week) or competitive sports (4-7 times/week). All patients underwent clinical and magnetic resonance imaging evaluation preoperative and 6, 18, and 36 months after autologous chondrocyte implantation. RESULTS: Group I patients showed significantly better results (< .01) in the International Cartilage Repair Society and Cincinnati scores than group II patients. Preoperative evaluation revealed no correlation between the sports activity levels and the clinical scores (P > .05). However, from the sixth month on, correlation was statistically significant, increasing from 6 to 18 months, and from 18 to 36 months postoperatively. CONCLUSION: Physical training improves long-term results after autologous chondrocyte implantation of the knee and should be carried out for at least 2 years after surgery. PMID- 17405885 TI - Lumbar intervertebral disk degeneration in elite competitive swimmers: a case control study. AB - BACKGROUND: The majority of orthopaedic problems experienced by competitive swimmers are related to pain in the shoulder, low back, and knee. Three of 39 national swim team members were hampered in their performance due to lumbar disk herniation at an international competition in 2001. There has been no previous research into lumbar disk degeneration in elite competitive swimmers. HYPOTHESIS: Excessive competitive swimming activities accelerate lumbar disk degeneration. STUDY DESIGN: Case control study; Level of evidence, 3. METHODS: Fifty-six elite swimmers (high-load group, 35 men and 21 women; mean age, 19.6 years) and a control group of 38 university recreational level swimmers (low-load group, 24 men and 14 women; mean age, 21.1 years) were evaluated for lumbar disk degeneration using magnetic resonance imaging. We compared the prevalence of disk degeneration and the disk level between the 2 groups and further investigated the relationship among their symptoms, swimming styles, and disk degeneration. RESULTS: Thirty-eight (68%) elite swimmers and 11 (29%) controls had degenerated disks at various disk levels, and the prevalence was significantly greater in the elite swimmers (P = .0002). Comparison between the 2 groups of the prevalence of disk degeneration at each level revealed that the disk level of L5-S1 was significantly more frequently degenerated in the high-load group (P = .026). There was no significant relationship observed among the variables of low back pain symptoms, swimming strokes, and disk degeneration. CONCLUSION: Excessive competitive swimming activities might exaggerate lumbar intervertebral disk degeneration, especially in the L5-S1 intervertebral segment. PMID- 17405886 TI - Comparison of preseason, midseason, and postseason neurocognitive scores in uninjured collegiate football players. AB - BACKGROUND: College football players sustain an average of 3 subconcussive blows to the head per game. Concussions correlate with decreases in standardized neurocognitive test scores. It is not known whether repetitive, subconcussive microtrauma associated with participation in a full season of collision sport affects neurocognitive test scores. HYPOTHESIS: No difference exists between preseason, midseason, and postseason Standardized Assessment of Concussion (SAC) and Immediate Post-Concussion Assessment and Cognitive Testing (ImPACT) scores when collegiate football players sustain subconcussive microtrauma from forceful, repetitive contact activity. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Fifty-eight members of a Division III collegiate football team who had no known concussion during the season voluntarily completed the SAC and ImPACT instruments preseason, midseason, and postseason. A repeated measures analysis of variance was used to compare the scores at the 3 time intervals (P < .05). RESULTS: No statistically significant decreases were found in overall SAC or ImPACT scores or in any of the domains or composites of the tests (P < .05) when preseason, midseason, and postseason scores were evaluated. CONCLUSIONS: ImPACT and SAC neurocognitive test scores are not significantly altered by a season of repetitive contact in collegiate football athletes who have not sustained a concussion. CLINICAL RELEVANCE: A diminution in SAC or ImPACT scores in concert with clinical symptoms and findings should be interpreted as evidence of a postconcussive event. PMID- 17405887 TI - Bruce Chabner awarded the Bob Pinedo Prize: a celebration of translational research and patient care excellence. PMID- 17405888 TI - The FDA in 2006: reasons for optimism. PMID- 17405889 TI - The Biomarkers Consortium: public and private sectors working in partnership to improve the public health. PMID- 17405890 TI - Achievements in systemic therapies in the pregenomic era in metastatic breast cancer. AB - Over the last decades, the introduction of several new agents into clinical practice has significantly improved disease control and obtained some, albeit rare, survival benefits in metastatic breast cancer (MBC). Despite these results, the choice of treatment for the majority of patients is still empirically based, since the only two predictive factors with level 1 evidence for clinical use are hormonal receptor status for endocrine therapy and HER-2 status for trastuzumab therapy. Important improvements in the endocrine therapy of both pre- and postmenopausal women with hormone-responsive disease have been achieved. For premenopausal women, ovarian function suppression with luteinizing hormone releasing hormone analogs combined with tamoxifen has become the standard treatment, although aromatase inhibitors plus ovarian function suppression are under evaluation. In postmenopausal patients, aromatase inhibitors have proved to be superior to standard endocrine therapies in either first- or second-line treatment and a novel antiestrogen compound, fulvestrant, has been introduced in clinical practice. Chemotherapy remains the treatment of choice for hormone unresponsive or resistant patients. Anthracyclines and taxanes have been used either alone or in combination as first-line chemotherapy, but with the more frequent use of these agents in the adjuvant setting, new standards are needed for first-line chemotherapy, and new and more efficacious treatments are required. In the subgroup of patients with tumors that overexpress HER-2, the use of trastuzumab alone or in combination with chemotherapy has modified the natural history of these tumors, even if only about one out of two patients obtains a clinical response. In this review we summarize the main achievements and the currently available treatment options for patients with MBC. PMID- 17405891 TI - Targeting the microtubules in breast cancer beyond taxanes: the epothilones. AB - Microtubule-targeting agents such as the taxanes are highly active against breast cancer and have become a cornerstone in the treatment of patients with early and advanced breast cancer. The natural epothilones and their analogs are a novel class of microtubule-stabilizing agents that bind tubulin and result in apoptotic cell death. Among this family of compounds, patupilone, ixabepilone, BMS-310705, ZK-EPO, and KOS-862 are in clinical development. Extensive preclinical studies have shown that epothilones are working through partially nonoverlapping mechanisms of action with taxanes. In the clinic, epothilones have been found in a series of phase I and phase II studies to be active even in patients who had recently progressed to taxanes. The toxicity profile of these agents consists mostly of sensory neuropathy, sometimes reversible. Neoadjuvant studies with epothilones have been conducted and a number of phase III studies in advanced breast cancer are either under way or have been recently completed. The results of these studies are eagerly awaited and it is anticipated that epothilones may become an important treatment option in patients with breast cancer. PMID- 17405892 TI - The emerging role of targeted therapy for hematologic malignancies: update on bortezomib and tipifarnib. AB - As therapy for hematologic malignancy evolves, new regimens and novel agents that target specific cellular processes allow a more optimistic prognosis for many patients. Bortezomib and tipifarnib are two new, targeted treatments for hematologic malignancies. Bortezomib, a proteasome inhibitor, has shown impressive efficacy in patients with relapsed multiple myeloma and as initial treatment, including before autologous stem cell transplantation. It has been studied as monotherapy and in combination with standard treatments such as dexamethasone, and with newer agents such as the immunomodulators thalidomide and lenalidomide; response is encouraging, even in patients who have relapsed after previously receiving components of a regimen as single agents. Bortezomib is generally well tolerated, including in combination with novel and conventional agents. Tipifarnib is a specific inhibitor of farnesyltransferase. Clinical trials in patients with high-risk acute leukemias and myelodysplastic syndromes have demonstrated good efficacy with tipifarnib. Continued investigation with these new, targeted treatments will further define their use as treatment options in patients with hematologic cancer. PMID- 17405893 TI - Medicinal cannabis does not influence the clinical pharmacokinetics of irinotecan and docetaxel. AB - OBJECTIVE: To date, data regarding the potential of cannabinoids to modulate cytochrome P450 isozyme 3A (CYP3A) activity are contradictory. Recently, a standardized medicinal cannabis product was introduced in The Netherlands. We anticipated an increased use of medicinal cannabis concurrent with anticancer drugs, and undertook a drug-interaction study to evaluate the effect of concomitant medicinal cannabis on the pharmacokinetics of irinotecan and docetaxel, both subject to CYP3A-mediated biotransformation. PATIENTS AND METHODS: Twenty-four cancer patients were treated with i.v. irinotecan (600 mg, n = 12) or docetaxel (180 mg, n = 12), followed 3 weeks later by the same drugs concomitant with medicinal cannabis (200 ml herbal tea, 1 g/l) for 15 consecutive days, starting 12 days before the second treatment. Blood samples were obtained up to 55 hours after dosing and analyzed for irinotecan and its metabolites (SN 38, SN-38G), respectively, or docetaxel. Pharmacokinetic analyses were performed during both treatments. Results are reported as the mean ratio (95% confidence interval [CI]) of the observed pharmacokinetic parameters with and without concomitant medicinal cannabis. RESULTS: Medicinal cannabis administration did not significantly influence exposure to and clearance of irinotecan (1.04; CI, 0.96-1.11 and 0.97; CI, 0.90-1.05, respectively) or docetaxel (1.11; CI, 0.94 1.28 and 0.95; CI, 0.82-1.08, respectively). CONCLUSION: Coadministration of medicinal cannabis, as herbal tea, in cancer patients treated with irinotecan or docetaxel does not significantly influence the plasma pharmacokinetics of these drugs. The evaluated variety of medicinal cannabis can be administered concomitantly with both anticancer agents without dose adjustments. PMID- 17405894 TI - Is molecular profiling ready for use in clinical decision making? AB - Molecular profiling, the classification of tissue or other specimens for diagnostic, prognostic, and predictive purposes based on multiple gene expression, is a technology that holds major promise for optimizing the management of patients with cancer. However, the use of these tests for clinical decision making presents many challenges to overcome. Assay development and data analysis in this field have been largely exploratory, and leave numerous possibilities for the introduction of bias. Standardization of profiles remains the exception. Classifier performance is usually overinterpreted by presenting the results as p-values or multiplicative effects (e.g., relative risks), while the absolute sensitivity and specificity of classification remain modest at best, especially when tested in large validation samples. Validation has often been done with suboptimal attention to methodology and protection from bias. The postulated classifier performance may be inflated compared to what these profiles can achieve. With the exception of breast cancer, we have little evidence about the incremental discrimination that molecular profiles can provide versus classic risk factors alone. Clinical trials have started to evaluate the utility of using molecular profiles for breast cancer management. Until we obtain data from these trials, the impact of these tests and the net benefit under real-life settings remain unknown. Optimal incorporation into clinical practice is not straightforward. Finally, cost-effectiveness is difficult to appreciate until these other challenges are addressed. Overall, molecular profiling is a fascinating and promising technology, but its incorporation into clinical decision making requires careful planning and robust evidence. PMID- 17405895 TI - Oral glutamine is effective for preventing oxaliplatin-induced neuropathy in colorectal cancer patients. AB - Oxaliplatin is effective in the treatment of metastatic colorectal cancer (MCRC) patients; however, severe neurotoxicity develops frequently. To assess the efficacy of oral glutamine for preventing neuropathy induced by oxaliplatin, a pilot study was performed. A total of 86 patients with MCRC treated at Taipei Veterans General Hospital were enrolled. Oxaliplatin (85 mg/m(2), days 1 and 15) plus weekly bolus 5-fluorouracil (5-FU; 500 mg/m(2)) and folinic acid (FA; 20 mg/m(2)) on days 1, 8, and 15 were given every 28 days as first-line treatment. Patients were randomized to receive (glutamine group; n = 42) or not receive (control group; n = 44) glutamine (15 g twice a day for seven consecutive days every 2 weeks starting on the day of oxaliplatin infusion). Efficacy of chemotherapy, neurological toxicity, and electrophysiological alterations were assessed. A lower percentage of grade 1-2 peripheral neuropathy was observed in the glutamine group (16.7% versus 38.6%) after two cycles of treatment, and a significantly lower incidence of grade 3-4 neuropathy was noted in the glutamine group after four cycles (4.8% versus 18.2%) and six cycles (11.9% versus 31.8%). By adding glutamine, interference with activities of daily living was lower (16.7% versus 40.9%), and need for oxaliplatin dose reduction was lower (7.1% versus 27.3%). There were no significant between-group differences in response to chemotherapy (52.4% versus 47.8%), electrophysiological abnormalities, grade 3-4 non-neurological toxicities (26.2% versus 22.8%), or survival. These data indi cate that oral glutamine significantly reduces the incidence and severity of peripheral neuropathy of MCRC patients receiving oxaliplatin without affecting response to chemotherapy and survival. PMID- 17405896 TI - A case of an alpha-fetoprotein-producing intrahepatic cholangiocarcinoma suggests probable cancer stem cell origin. AB - Recent evidence suggests that some cancers may originate from cancer stem cells, which may derive from carcinogenesis of normal stem cells. A hepatic progenitor cell population, which gives rise to hepatocytes and cholangiocytes, has been suggested in humans, though whether these cells can give rise to malignant tumors has not been confirmed. We report here a case of an alpha-fetoprotein (AFP) producing intrahepatic cholangiocarcinoma (ICC) in an 81-year-old woman with chronic hepatitis C viral infection, suggesting malignant transformation of hepatic stem cells as a mechanism for hepatic neoplasia. Abdominal computed tomography revealed a low-density mass with surrounding enhancement measuring 5 cm x 5 cm in segments IV and VIII of the liver. The preoperative serum levels of tumor markers were 1.7 ng/ml of carcinoembryonic antigen, 22 mAU/ml of protein induced by vitamin K absence or antagonist II, 43.4 U/ml of carbohydrate antigen 19-9, and 1,560 ng/ml of AFP. Following central bisegmentectomy of the liver, serum AFP levels decreased dramatically. Histologically, the tumor cells showed indistinct glandular structures with abundant fibrous stroma. Immunohistochemical analysis demonstrated that the neoplastic cells reacted strongly to antibodies against AFP and cytokeratin (CK) 7. In addition, cancer cells showed partially positive reaction to anti-CK14, a liver stem cell marker, and to anticluster designation (CD) 133, a hematopoietic stem cell marker, and negative reaction to antihepatocyte paraffin (HepPar) 1. These data may indicate that the tumor was derived from a normal liver stem cell that underwent oncogenic transformation. PMID- 17405897 TI - Second-generation epidermal growth factor receptor tyrosine kinase inhibitors in non-small cell lung cancer. AB - Inhibiting epidermal growth factor receptor (EGFR) signaling has proven to be an effective strategy for treating non-small cell lung cancer (NSCLC) patients and the first generation of agents developed for this purpose, gefitinib and erlotinib, stimulated a unique escalation in both biologic and clinical research within the field. Second-generation EGFR-targeted agents that aim to further improve patient outcomes are now in preclinical and clinical trials. This review discusses four promising agents that are currently being studied in NSCLC: EKB 569, HKI-272, CI-1033, and ZD6474. PMID- 17405898 TI - Optimal adjuvant therapy for non-small cell lung cancer--how to handle stage I disease. AB - The standard of care for resected stage II-IIIA non-small cell lung cancer (NSCLC) now includes adjuvant chemotherapy based on the results of three phase III studies using cisplatin-based regimens--the International Adjuvant Lung Trial, the National Cancer Institute of Canada JBR.10 trial, and the Adjuvant Navelbine International Trialist Association trial. The role of adjuvant chemotherapy for stage I disease remains controversial. A recent meta-analysis (the Lung Adjuvant Cisplatin Evaluation) showed potential harm with the addition of adjuvant cisplatin for stage IA disease and no survival benefit for this modality in stage IB disease. Updated results from the Cancer and Leukemia Group B 9633 trial, the only trial to focus exclusively on stage IB patients, no longer show a statistically significant survival benefit from adjuvant chemotherapy in this population, except for the subgroup of patients with larger tumors. It may be that trials have been underpowered to detect a small benefit for patients with stage IB disease, or there may really not be benefit to adding adjuvant therapy for this stage of disease. Additional markers, such as tumor size or the presence or absence of certain tumor proteins like ERCC1, may help to determine which patients with resected stage I NSCLC may benefit from adjuvant chemotherapy. Strategies such as inhibition of angiogenesis pathways and the epidermal growth factor receptor are under exploration. PMID- 17405899 TI - Safe administration of iodine-131 tositumomab after repeated infusion-related reactions to rituximab. AB - Infusion-related reactions during administration of monoclonal antibody therapy are often mild and unlikely to recur with subsequent treatment. If patients experience another severe reaction upon reattempting treatment, future treatments with the same agent are typically not pursued. It is unclear whether different monoclonal antibodies that bind the same tumor cell or antigen are likely to induce similar infusion reactions. Here, we report the case of a patient with repeated severe infusion reactions with rituximab who subsequently safely received treatment with iodine-131 tositumomab and discuss the relevant literature. PMID- 17405900 TI - Prognostic factors and risk-based therapy in pediatric acute myeloid leukemia. AB - Acute myeloid leukemia (AML) has posed significant therapeutic challenges to pediatric oncologists. Despite intensive therapy, half of the children with AML relapse and die from their disease. Efforts to identify risk factors in AML are directed toward defining populations who may benefit from alternative therapies. Patients at lower risk for relapse may benefit from treatment de-escalation, sparing them adverse side effects. Management of high-risk patients may prove more difficult, as the nearly myeloablative nature of AML therapy leaves little room for therapy escalation short of stem cell transplantation. This review evaluates prognostic factors in pediatric AML and discusses the feasibility of using these factors in risk-adapted therapy regimens. PMID- 17405901 TI - FDA drug approval summary: bevacizumab plus FOLFOX4 as second-line treatment of colorectal cancer. AB - On June 20, 2006, the U.S. Food and Drug Administration (FDA) approved bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA), administered in combination with FOLFOX4 (5-fluorouracil, leucovorin, and oxaliplatin) for the second-line treatment of metastatic carcinoma of the colon or rectum. Efficacy and safety were demonstrated in one Eastern Cooperative Oncology Group (ECOG) open-label, multicenter, randomized, three-arm, active-controlled trial enrolling 829 adult patients. Patients had received a fluoropyrimidine- and irinotecan based regimen as initial therapy for metastatic disease; or they had received prior adjuvant irinotecan-based chemotherapy and had recurred within 6 months of completing therapy. Treatments included bevacizumab, 10 mg/kg, as a 90-minute i.v. infusion on day 1, every 2 weeks, either alone or in combination with FOLFOX4, or FOLFOX4 alone. The bevacizumab monotherapy arm was closed to accrual after an interim efficacy analysis suggested a possibly shorter survival in that arm. Overall survival (OS), the primary study endpoint, was significantly longer for patients receiving bevacizumab in combination with FOLFOX4 than for those receiving FOLFOX4 alone. The objective response rate was significantly higher in the FOLFOX4 plus bevacizumab arm than in the FOLFOX4 alone arm. The duration of response was approximately 6 months for both treatment arms. Patients treated with the bevacizumab combination were also reported, based on investigator assessment, to have significantly longer progression-free survival. There were no new bevacizumab safety signals. The most serious, and sometimes fatal, bevacizumab toxicities are gastrointestinal perforation, wound-healing complications, hemorrhage, arterial thromboembolic events, hypertensive crisis, nephrotic syndrome, and congestive heart failure. PMID- 17405902 TI - Transforming growth factor beta is a critical regulator of adult human islet plasticity. AB - Tissue plasticity is well documented in the context of pancreatic regeneration and carcinogenesis, with recent reports implicating dedifferentiated islet cells both as endocrine progenitors and as the cell(s) of origin in pancreatic adenocarcinoma. Accordingly, it is noteworthy that accumulating evidence suggests that TGFbeta signaling is essential to pancreatic endocrine development and maintenance, whereas its loss is associated with the progression to pancreatic adenocarcinoma. The aim of this study was to examine the role of TGFbeta in an in vitro model of islet morphogenetic plasticity. Human islets were embedded in a collagen gel and cultured under conditions that induced transformation into duct like epithelial structures (DLS). Addition of TGFbeta caused a dose-dependent decrease in DLS formation. Although it was demonstrated that collagen-embedded islets secrete low levels of TGFbeta, antibody-mediated neutralization of this endogenously released TGFbeta improved DLS formation rates, suggesting local TGFbeta concentrations may in fact be higher. Time course studies indicated that TGFbeta signaling was associated with an increase in ERK and p38 MAPK phosphorylation, although inhibitor-based studies were consistent with an islet endocrine-stabilizing effect mediated by p38 alone. Localization of TGFbeta signaling molecules suggested that the action of TGFbeta is directly on the beta cell to inhibit apoptosis and thus stabilize endocrine phenotype. PMID- 17405903 TI - The angiostatic 16K human prolactin overcomes endothelial cell anergy and promotes leukocyte infiltration via nuclear factor-kappaB activation. AB - The 16-kDa N-terminal fragment of human prolactin (16K hPRL) is a potent angiostatic factor that inhibits tumor growth in mouse models. Using microarray experiments, we have dissected how the endothelial-cell genome responds to 16K hPRL treatment. We found 216 genes that show regulation by 16K hPRL, of which a large proportion turned out to be associated with the process of immunity. 16K hPRL induces expression of various chemokines and endothelial adhesion molecules. These expressions, under the control of nuclear factor-kappaB, result in an enhanced leukocyte-endothelial cell interaction. Furthermore, analysis of B16-F10 tumor tissues reveals a higher expression of adhesion molecules (intercellular adhesion molecule 1, vascular cell adhesion molecule 1, or E-selectin) in endothelial cells and a significantly higher number of infiltrated leukocytes within the tumor treated with 16K hPRL compared with the untreated ones. In conclusion, this study describes a new antitumor mechanism of 16K hPRL. Because cellular immunity against tumor cells is a crucial step in therapy, the discovery that treatment with 16K hPRL overcomes tumor-induced anergy may become important for therapeutic perspectives. PMID- 17405904 TI - Liver X receptor stimulates cholesterol efflux and inhibits expression of proinflammatory mediators in human airway smooth muscle cells. AB - Human (h) airway smooth muscle (ASM) cells are important mediators of the inflammatory process observed in asthma and other respiratory diseases. We show here that primary hASM cells express liver X receptor (LXR; alpha and beta subtypes), an oxysterol-activated nuclear receptor that controls expression of genes involved in lipid and cholesterol homeostasis, and inflammation. LXR was functional as determined by transient assays using LXR-responsive reporter genes and by analysis of mRNA and protein expression of endogenous LXR target genes in cells exposed to LXR agonists. LXR activation induced expression of the ATP binding cassette transporters ABCA1 and ABCG1 and increased efflux of cholesterol to apolipoprotein AI and high-density lipoprotein acceptors, pointing to a role for hASM cells in modulating cholesterol homeostasis in the airway. Under inflammatory conditions, hASM cells release a variety of chemokines and cytokines that contribute to inflammatory airway diseases. Activation of LXR inhibited the expression of multiple cytokines in response to proinflammatory mediators and blocked the release of both granulocyte macrophage colony-stimulating factor and granulocyte colony stimulating factor. LXR activation also inhibited proliferation of hASM cells and migration toward platelet-derived growth factor chemoattractant, two important processes that contribute to airway remodeling. Our findings reveal biological roles for LXR in ASM cells and suggest that modulation of LXR activity offers prospects for new therapeutic approaches in the treatment of asthma and other inflammatory respiratory diseases. PMID- 17405905 TI - Direct interactions with G alpha i and G betagamma mediate nongenomic signaling by estrogen receptor alpha . AB - Estrogen induces G protein-dependent nongenomic signaling in a variety of cell types via the activation of a plasma membrane-associated subpopulation of estrogen receptor alpha (ER alpha). Using pull-down experiments with purified recombinant proteins, we now demonstrate that ER alpha binds directly to G alpha i and G betagamma. Mutagenesis and the addition of blocking peptide reveals that this occurs via amino acids 251-260 and 271-595 of ER alpha, respectively. Studies of ER alpha complexed with heterotrimeric G proteins further show that estradiol causes the release of both G alpha i and G betagamma without stimulating GTP binding to G alpha i. Moreover, in COS-7 cells, the disruption of ER alpha-G alpha i interaction by deletion mutagenesis of ER alpha or expression of blocking peptide, as well as G betagamma sequestration with beta-adrenergic receptor kinase C terminus, prevents nongenomic responses to estradiol including src and erk activation. In endothelial cells, the disruption of ER alpha-G alpha i interaction prevents estradiol-induced nitric oxide synthase activation and the resulting attenuation of monocyte adhesion that contributes to estrogen-related cardiovascular protection. Thus, through direct interactions, ER alpha mediates a novel mechanism of G protein activation that provides greater diversity of function of both the steroid hormone receptor and G proteins. PMID- 17405906 TI - NF-kappaB2 mutation targets TRAF1 to induce lymphomagenesis. AB - The NF-kappaB2 gene is recurrently mutated in human lymphoid malignancies. However, a causal relationship between NF-kappaB2 mutation and lymphomagenesis has not been established. It is also unclear how the mutation may lead to lymphoid malignancies. We report the generation of transgenic mice with targeted expression of p80HT, a lymphoma-associated NF-kappaB2 mutant, in lymphocytes. The transgenic mice display a marked expansion of peripheral B cell populations and develop predominantly small B cell lymphomas. p80HT expression has no apparent effect on the proliferation of B cells, but renders them specifically resistant to apoptosis induced by cytokine deprivation and mitogenic stimulation. Lymphocytes and lymphoma cells from p80HT mice express high levels of TRAF1, an antiapoptotic protein also implicated in lymphoid malignancies. p80HT binds the TRAF1 promoter in vivo and activates TRAF1 transcription. Moreover, TRAF1 knockdown abrogates the antiapoptotic activity of p80HT and TRAF1 deficiency reestablishes B cell homeostasis in p80HT mice. These findings demonstrate NF kappaB2 mutation as an oncogenic event in vivo and suggest a molecular pathway for TRAF1 activation in the pathogenesis of lymphomas. PMID- 17405907 TI - Kinase domain mutations of BCR-ABL frequently precede imatinib-based therapy and give rise to relapse in patients with de novo Philadelphia-positive acute lymphoblastic leukemia (Ph+ ALL). AB - Acquired imatinib resistance in advanced Philadelphia-positive acute lymphoblastic leukemia (Ph(+) ALL) has been associated with mutations in the kinase domain (KD) of BCR-ABL. We examined the prevalence of KD mutations in newly diagnosed and imatinib-naive Ph(+) ALL patients and assessed their clinical relevance in the setting of uniform frontline therapy with imatinib in combination with chemotherapy. Patients enrolled in the German Multicenter Study Group for Adult Acute Lymphoblastic Leukemia (GMALL) trial ADE10 for newly diagnosed elderly Ph(+) ALL were retrospectively examined for the presence of BCR ABL KD mutations by denaturing high-performance liquid chromatography (D-HPLC), cDNA sequencing, and allele-specific polymerase chain reaction (PCR). A KD mutation was detected in a minor subpopulation of leukemic cells in 40% of newly diagnosed and imatinib-naive patients. At relapse, the dominant cell clone harbored an identical mutation in 90% of cases, the overall prevalence of mutations at relapse was 80%. P-loop mutations predominated and were not associated with an inferior hematologic or molecular remission rate or shorter remission duration compared with unmutated BCR-ABL. BCR-ABL mutations conferring high-level imatinib resistance are present in a substantial proportion of patients with de novo Ph(+) ALL and eventually give rise to relapse. This provides a rationale for the frontline use of kinase inhibitors active against these BCR-ABL mutants. PMID- 17405908 TI - Antigen-activated human T lymphocytes express cell-surface NKG2D ligands via an ATM/ATR-dependent mechanism and become susceptible to autologous NK- cell lysis. AB - Recent evidence indicates that natural killer (NK) cells can negatively regulate T-cell responses, but the mechanisms behind this phenomenon as a consequence of NK-T-cell interactions are poorly understood. We studied the interaction between the NKG2D receptor and its ligands (NKG2DLs), and asked whether T cells expressed NKG2DLs in response to superantigen, alloantigen, or a specific antigenic peptide, and if this rendered them susceptible to NK lysis. As evaluated by FACS, the major histocompatibility complex (MHC) class I chain-related protein A (MICA) was the ligand expressed earlier on both CD4(+) and CD8(+) T cells in 90% of the donors tested, while UL16-binding protein-1 (ULBP)1, ULBP2, and ULBP3 were induced at later times in 55%-75% of the donors. By carboxyfluorescein diacetate succinimidyl ester (CFSE) labeling, we observed that NKG2DLs were expressed mainly on T cells that had gone through at least one division. Real-time reverse transcription polymerase chain reaction confirmed the expression of all NKG2DLs, except ULBP4. In addition, T-cell activation stimulated phosphorylation of ataxia telangiectasia mutated (ATM), a kinase required for NKG2DLs expression after DNA damage, and ATM/Rad3-related kinase (ATR) inhibitors blocked MICA induction on T cells with a mechanism involving NF-kappaB. Finally, we demonstrated that activated T cells became susceptible to autologous NK lysis via NKG2D/NKG2DLs interaction and granule exocytosis, suggesting that NK lysis of T lymphocytes via NKG2D may be an additional mechanism to limit T-cell responses. PMID- 17405910 TI - Targeting bacterial endotoxin: two sides of a coin. AB - The term sepsis describes a potentially lethal clinical condition that develops as a result of a dysregulated host response to bacterial infection. The most common bacterial component implicated in initiating the septic syndrome is a cell wall molecule derived from Gram-negative bacteria, known as lipopolysaccharide (LPS) or endotoxin. Like all mammals, humans are equipped with an LPS-sensing machinery consisting, primarily, of LPS-binding protein (LBP), CD14, a glycosylphosphatidylinositol (GPI)-anchored monocyte differentiation antigen, and toll-like receptor 4 (TLR4), a signal-transducing integral membrane protein. Modest stimulation of TLR4 facilitates the elimination of invading microorganisms. Potent TLR4 stimulation, however, produces severe reactions in the host, often leading to multiple organ failure and death. The search for pharmaceuticals that reduce mortality in septic patients has been a painstaking process. Thus far, only a few compounds have been found to significantly reduce mortality rates. Perhaps one of the more promising therapeutic strategies currently pursued is based on the identification of synthetic or naturally occurring substances that neutralize LPS or inhibit LPS-mediated activation of host immune cells, such as monocytes and macrophages. Here, we describe a number of diverse molecular structures with a capacity to either enhance or blunt LPS induced monocyte activation. The underlying molecular mechanisms are discussed. PMID- 17405911 TI - Interaction between the Helicobacter pylori CagA and alpha-Pix in gastric epithelial AGS cells. AB - The gastric pathogen Helicobacter pylori (H. pylori) translocates the CagA protein into epithelial cells by a type IV secretion process. Upon translocation into the host cell cytosol, CagA undergoes tyrosine phosphorylation. Phosphorylation of CagA occurs within the C terminus of the protein and is mediated by members of the Src family of tyrosine kinases. Phosphorylation of CagA induces the dephosphorylation of as yet unidentified cellular proteins, rearrangements of the host cell actin cytoskeleton, and cell scattering. This article aims to determine the cellular protein that interacts with CagA. Gastric epithelial AGS cells were stimulated with CagA-positive H. pylori (NCTC11637, at a bacteria/cell ratio of 500:1) and cultured in antibiotic-free medium. Proteins were isolated from the cells with or without H. pylori infection. CagA interactive protein was determined by immunoprecipitation using anti-CagA antibody and proteomic analysis. We found that alpha-Pix interacts with CagA and alpha-Pix was constitutively expressed in AGS cells. Upon H. pylori stimulation, CagA was translocated into the cells and the expression of alpha-Pix (PAK interactive exchange factor) was increased in AGS cells time dependently. The interaction of alpha-Pix with CagA was increased by H. pylori infection in AGS cells. Phosphorylation of CagA induces the dephosphorylation of alpha-Pix in AGS cells. alpha-Pix is a family of PAK-binding proteins that strongly activates PAK (p21-activated tyrosine kinase). PAK regulates changes in gene expression and mediates actin cytoskeletal and cell morphological changes. The novel finding of this study is that phosphorylation of CagA induces the dephosphorylation of alpha Pix, which may modulate cytoskeletal changes of gastric epithelial cells through PAK. PMID- 17405912 TI - Expression of suppressors of cytokine signaling-3 in Helicobacter pylori-infected rat gastric mucosal RGM-1cells. AB - Our previous studies show that Helicobacter pylori (H. pylori) induces oxidative stress and the expression of proinflammatory cytokines in gastric epithelial cells. H. pylori induces the expression of molecular chaperones and proteins involved in protein-folding machinery as a defense mechanism against cellular stress. The suppressors of cytokine signaling (SOCS) are known as negative regulators of major immune signal pathways. The purpose of this article is to determine whether H. pylori in a Korean isolate (HP99) induces the expression of SOCS in rat gastric mucosal RGM-1 cells as a defense mechanism. As a result, HP99 induced SOCS-3 expression time-dependently in RGM-1 cells. SOCS-1 was not expressed while SOCS-2 expression was not changed by HP99 infection in RGM-1 cells. SOCS-3 might have a defensive role in H. pylori-infected gastric mucosal cells. Further study by manipulating SOCS-3 gene should be performed to investigate the physiological meaning of SOCS-3 induced by H. pylori in gastric mucosal cells. PMID- 17405913 TI - Role of proteinase-activated receptor-2 on cyclooxygenase-2 expression in H. pylori-infected gastric epithelial cells. AB - Proteinase-activated receptor-2 (PAR-2) belongs to a novel subfamily of G protein coupled receptors with seven-transmembrane domains. PAR-2 is activated by serine proteases, such as trypsin, mast cell tryptase, and allergic or bacterial proteases. The presence of trypsin has been shown in human stomach. Cyclooxygenase-2 (COX-2) is induced by inflammatory cytokines, growth factors, gastrin, and reactive oxygen species in gastric epithelial cells, which may lead to mutagenesis and subsequent metaplasia, dysplasia, and cancer formation. We investigated whether PAR-2 is activated in H. pylori (HP99)-infected cells, which is related to COX-2 induction in gastric epithelial cells. After treatment of H. pylori to AGS (gastric adenocarcinoma) cells at a bacteria/cell ratio of 100:1, we determine the expression and the activation of PAR-2 and the expression of COX 2. The same experiments were performed in the cells treated with PAR-2 agonist peptide. mRNA and protein expression of PAR-2 and COX-2 were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. PAR-2 activation was assessed by increase in intracellular calcium level. As a result, H. pylori induced the activation and expression of PAR-2 as well as COX-2 expression. PAR-2 agonist peptide augmented H. pylori-induced COX-2 expression in AGS cells. H. pylori induces COX-2 expression, which is mediated by both activation and expression of PAR-2 in gastric epithelial cells. PMID- 17405914 TI - Control of human herpes virus type 8-associated diseases by NK cells. AB - The "natural killer" (NK) cells preferentially kill targets lacking surface major histocompatibility complex class I (MHC-I) molecule expression. NK cells recognize these targets through membrane receptors, which can trigger activating or inhibitory signals for killing. Several tumors or virus-infected cells downregulate MHC-I expression as a mechanism to evade recognition and killing by cytotoxic T lymphocytes (CTL). They, however, become targets for NK cells cytotoxic activity. NK cell activity is reduced during disease progression in human immunodeficiency virus (HIV) infection, and in individuals with AIDS associated tumors linked with infection by the oncogenic human herpes virus type 8 (HHV8), including Kaposi's sarcoma (KS) and primary effusion lymphomas (PEL). We have demonstrated that AIDS-related KS (AIDS-KS) is characterized by an increased expression of inhibitory receptors by T lymphocytes, and that HIV-non infected patients with KS (classic KS, C-KS) have a substantial number of NK cells bearing these same receptors. NK cells from patients with C-KS are normally equipped with cytolytic molecules including granzyme A and perforin. However, the cytotoxic activity of NK cells is reduced in patients with C-KS, AIDS-KS, or PEL patients, who are all infected by the HHV8, and this correlates with disease severity. Moreover, we have found that HHV8-infected cell lines established from PELs have a reduced surface expression of MHC-I molecules and are sensitive to the lysis mediated by NK cells. Since PEL cells express the same HHV8 latency program as KS cells, these data point to MHC-I downregulation by HHV8 as a primary immune evasion mechanism against CTL responses, further reinforced by upregulation of inhibitory receptors on T and NK cells in the setting of HIV and/or HHV8 infection. Thus, studies on killing receptor regulation and signaling in T and NK cells may shed light on the pathogenesis of HHV8-associated tumors both in HIV-infected or -noninfected patients. PMID- 17405915 TI - Analysis of tissue distribution of TNF-alpha, TNF-alpha-receptors, and the activating TNF-alpha-converting enzyme suggests activation of the TNF-alpha system in the aging intervertebral disc. AB - We immunohistochemically analyzed the expression and localization of TNF-alpha, its receptors TNF-RI and -RII, and the TNF-alpha-activating enzyme TACE in human autopsy (n=63) and surgical (n=35) lumbar intervertebral disc samples. In parallel, the TNF-alpha-mRNA was quantified by reverse transcriptase-polymerase chain reaction (RT-PCR). All samples were morphometrically evaluated for the proportion of positively labeled cells in the different anatomical regions of the disc. We detected a significant and comparable expression of all four parameters beginning in young adult age (c. 18 years) and being most extensive in the nucleus pulposus. This level was slightly reduced in older age discs. The annulus fibrosus contained significantly less labeled cells. In accord, the number of TNF alpha-transcripts was elevated in most cases with immunohistochemical TNF-alpha expression. We provide clear evidence that TNF-alpha is expressed in discs of increasing age, which correlates with histomorphological signs of disc degeneration. In consequence, TNF-alpha seems to be activated (by the converting enzyme TACE) and biologically active through its receptors in human lumbar disc tissue. PMID- 17405916 TI - Upregulation of apolipoprotein B secretion, but not lipid, by tumor necrosis factor-alpha in rat hepatocyte cultures in the absence of extracellular fatty acids. AB - Tumor necrosis factor-alpha (TNF-alpha) plays a pivotal role in the host response to infection. Rapidly liberated to the bloodstream, TNF-alpha triggers the production of other cytokines and the acute-phase response. Hypertriglyceridemia is a sepsis hallmark associated with high plasma levels of very low-density lipoprotein (VLDL) particles, partly ascribed to increased hepatic production. The kinetics of the hepatocyte response, the cytokine/s responsible, and the underlying mechanisms are not fully elucidated. VLDL biogenesis is a complex, time-consuming process that depends on lipid availability and microsomal triglyceride transfer protein (MTP) activity for correct apolipoprotein B (apoB) lipidation. Studies were performed to define the direct effect of TNF-alpha on VLDL secretion rate and composition in rat hepatocytes cultured in conditions resembling the fed situation. Increases of 17-24% in the number of VLDL particles secreted and of 44-88% in the cellular levels of apoB mRNA were caused by 5, 20, or 100 ng/mL TNF-alpha in 8 h. Lipoprotein secretion returned to baseline levels in 16 h, whereas TNF-alpha-treated cells continued to exhibit higher apoB transcript levels. The mass of each lipid class in secreted VLDL and of MTP mRNA in cells was not affected by any of the tested TNF-alpha doses or treatment periods. These findings indicate that over a wide range of concentrations, TNF alpha was capable of inducing sustained upregulation of apoB mRNA expression and transient increase in secretion of its protein, but, apparently, VLDL triglyceride secretion was not a TNF-alpha target under conditions in which fatty acids were not extracellularly provided. PMID- 17405917 TI - Gene expression profiling of LPS-stimulated murine macrophages and role of the NF kappaB and PI3K/mTOR signaling pathways. AB - Lipopolysaccharide (LPS), a major component of the outer membrane of Gram negative bacteria, activates a broad spectrum of signaling pathways in immune cells. In this article, RAW264.7 cells have been stimulated for 4 h with 1 microg/mL of LPS in the presence or not of specific inhibitors of the NF-kappaB pathway (BAY 11-7082) and the PI3K pathway (LY294002). Gene expression profiles were characterized using the DNA microarray "Dual Chip Mouse Inflammation." This array monitors the expression of 233 genes encoding proteins playing a role in inflammation. Both signaling pathways exert an important role in the response to LPS, but they are not completely overlapping. For example, genes encoding the PAF receptor, PAI-1, PlA2 (group V), IL-13 receptor (alpha2), and GTP cyclohydrolase 1, were upregulated after LPS treatment, but this upregulation was counteracted by LY294002. The same was observed for BAY 11-7082: genes encoding the kit ligand, TLR2, or TNFRSF5 were mainly under the control of NF-kappaB. NF-kappaB plays an important role in the macrophage response to LPS, but we have also shown that the PI3K pathway partially contributes to it. Further experiments with the specific inhibitor of mTOR (rapamycin) will provide more information on the specific contribution of the PI3K/mTOR pathway in the inflammatory response in LPS-stimulated macrophages. PMID- 17405918 TI - Modification of proteins by cyclopentenone prostaglandins is differentially modulated by GSH in vitro. AB - Prostanoids with cyclopentenone structure (cyP) display a potent anti inflammatory and antiproliferative activity. CyP are reactive compounds, which may modulate cellular functions by multiple mechanisms, including the direct covalent modification of cysteine residues by Michael addition. This interaction displays selectivity since only a subset of cellular proteins is modified by cyP. Several factors have been proposed to influence the selectivity and/or extent of cyP addition to proteins, including determinants related to protein and cyP structure, and levels of cellular thiols, such as glutathione (GSH). Here we have explored the ability of biotinylated cyP analogs to modify several recombinant proteins in vitro, and the influence of GSH in these effects. We have observed that protein modification by cyP is protein- and cyP-selective. Under our conditions, biotinylated 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)-B) was more efficient than biotinylated PGA(1) (PGA(1)-B) at forming adducts with components of the transcription factors NF-kappaB and activator protein-1 (AP-1). However, both biotinylated cyP were nearly equipotent at modifying human GSTP1-1. Interestingly, the presence of GSH differentially modulated the formation of protein-cyP adducts. Under our conditions, GSH reduced the incorporation of cyP into GST, but improved their binding to p50, more intensely in the case of PGA(1) B. These results evidence the importance of GSH-cyP and/or GSH-protein interactions for the selectivity of protein modification by cyP and suggest a complex role for GSH that may be related to its ability to prevent protein oxidation or induce conformational alterations. This may shed light on the factors involved in the pleiotropic effects of electrophiles with therapeutic potential. PMID- 17405919 TI - Signaling pathways involved in proteinase-activated receptor1-induced proinflammatory and profibrotic mediator release following lung injury. AB - The proteinase-activated receptors (PAR1 to 4) represent important novel therapeutic targets in fibroproliferative lung disorders, linking signals from coagulation proteinases to the production of proinflammatory and profibrotic mediators, in response to lung injury. This review focuses on the current understanding of cytokine and chemokine release downstream of PAR1 activation in the context of fibroproliferative lung disease, with an emphasis on the induction of the chemokine CCL2/MCP-1. PMID- 17405920 TI - Signaling pathway used by HSV-1 to induce NF-kappaB activation: possible role of herpes virus entry receptor A. AB - We have previously demonstrated that wild-type herpes simplex virus type 1 (HSV 1), as well as nonreplicating UV-inactivated HSV-1, promptly activates the nuclear factor-kappaB (NF-kappaB) in U937 monocytoid cells and that glycoprotein D (gD) of HSV-1 is sufficient by itself to exert a similar effect. We then investigated the signaling pathway used by HSV-1 to initiate NF-kappaB activation and, particularly, whether our observation could be related to the capability of HSV-1-gD to directly stimulate NF-kappaB through its interaction with the herpes virus entry receptor A (HveA). Here we report that: (a) co-cultivation of U937 cells with an adherent cell line expressing wild-type gD on its surface led to increased NF-kappaB activation, while co-cultivation with the same adherent cell line expressing a mutated form of gD, lacking the capability to bind HveA, did not cause the same effect; (b) exposure to UV-inactivated HSV-1 induced the activation of NF-kappaB in HveA-expressing U937 and THP-1 cells, but not in non HveA-expressing HEp-2 cells; and (c) activation of NF-kappaB in U937 and THP-1 cells exposed to soluble gD was inhibited by an antibody able to interfere with gD-HveA interaction. These results suggest that HSV-1-gD-HveA interaction initiates a signal transduction pathway leading to NF-kappaB activation. PMID- 17405921 TI - Melphalan reduces the severity of experimental colitis in mice by blocking tumor necrosis factor-alpha signaling pathway. AB - Melphalan is an alkylating agent, which is commonly used as an antineoplastic drug. Its cytostatic effect can be realized in humans in the dose range of 0.6 1.4 mg/kg body weight. However, previously it was shown that in the case of gradual dose decrease, the number of targets for alkylation was also reduced and the drug lost its cytostatic properties switching to cell growth modifier. It has been postulated that application of alkylating agents in such ultra-low concentrations (50- to 100-fold lower than cytostatic ones) may result in a beneficial effect in the therapy of diseases associated with mucosa inflammation. The aim of the article was to investigate the effect of ultra-low doses of melphalan in the murine experimental colitis induced by the replacement of drinking water with 5% solution of dextran sulphate sodium (DSS). Daily administration of melphalan (25 microg/kg body weight) markedly reduced the severity of DSS-colitis as determined by clinical and quantitative histological criteria. Both systemic and local anti-inflammatory effects of melphalan have been observed. The possible mechanisms of the beneficial effect of the drug have been discussed. PMID- 17405922 TI - Cellular prion protein signaling in serotonergic neuronal cells. AB - The cellular prion protein PrP(C) is the normal counterpart of the scrapie prion protein PrP(Sc), the main component of the infectious agent of transmissible spongiform encephalopathies (TSEs). It is a ubiquitous cell-surface glycoprotein, abundantly expressed in neurons, which constitute the targets of TSE pathogenesis. Taking advantage of the 1C11 neuroectodermal cell line, endowed with the capacity to convert into 1C11(5-HT) serotonergic or 1C11(NE) noradrenergic neuronal cells, allowed us to ascribe a signaling function to PrP(C). Antibody-mediated ligation of PrP(C) recruits transduction pathways, which involve nicotinamide adenine dinucleotide phosphate (NADPH) oxidase dependent reactive oxygen species production and target the extracellular regulated kinases ERK1/2. In fully differentiated cells only, these effectors are under the control of a PrP(C)-caveolin-Fyn platform, located on neuritic extensions. In addition to its proper signaling activity, PrP(C) modulates the agonist-induced response of the three serotonergic G protein-coupled receptors present on the 1C11(5-HT) differentiated cells. The impact of PrP(C) ligation on the receptor couplings depends on the receptor subtype and the pathway considered. The implementation of the PrP(C)-caveolin complex again is mandatory for PrP(C) to exert its action on 5-HT receptor signaling. Our current data argue that PrP(C) interferes with the intensities and/or dynamics of G protein activation by agonist-bound 5-HT receptors. By mobilizing transduction cascades controlling the cellular redox state and the ERK1/2 kinases and by altering 5-HT receptor-mediated intracellular response, PrP(C) takes part in the homeostasis of serotonergic neuronal cells. These findings may have implications for future research aiming at understanding the fate of serotonergic neurons in prion diseases. PMID- 17405923 TI - Strongly reduced number of parvalbumin-immunoreactive projection neurons in the mammillary bodies in schizophrenia: further evidence for limbic neuropathology. AB - The mammillary bodies (MB) are important relay nuclei within limbic and extralimbic connections. They are known to play important roles in memory formation and are affected in alcoholism and vitamin B1 deficiency. Their strategic position linking temporo-limbic to cortico-thalamic brain structures make the MB a candidate brain structure for alterations in schizophrenia. We studied 15 postmortem brains of schizophrenics and 15 matched control brains. Brain sections were stained either with Heidenhain-Woelcke, glutamic acid decarboxylase (GAD), calretinin, or parvalbumin. We determined the volumes of the MB and performed cell countings using stereological principles and a computerized image analysis system. The volumes of MB do not differ between schizophrenics and controls. However, in schizophrenia the number of neurons as well as the resulting neuronal densities was significantly reduced on both sides (on left side by 38.9%, on right side by 22%). No changes were seen in the number of GAD expressing or calretinin-containing neurons, whereas the number of parvalbumin immunoreactive MB neurons was reduced by more than 50% in schizophrenia. This cell loss (as a result of developmental malformation and/or neurodegeneration) points to a prominent involvement of the MB in the pathomorphology of schizophrenia. Parvalbumin-immunoreactive GABAergic interneurons have been reported to be diminished in schizophrenia. However, in the MB parvalbumin labels a subpopulation of glutamate/aspartate-containing neurons projecting mainly to the anterior thalamus. Thus, our data provide new evidence for impaired limbic circuits in schizophrenia. PMID- 17405924 TI - Alterations of synaptic turnover rate in aging may trigger senile plaque formation and neurodegeneration. AB - The changes of synaptic ultrastructure were investigated by morphometry in the frontal (FC) and temporal (TC) cortex of adult and aged monkeys, to assess the potential role of age-related synaptic deterioration in neurodegeneration. The average synaptic size (S), the synaptic numeric density (Nv: number of synapses/microm(3) of tissue), the synaptic surface density (Sv: overall area of synaptic junctional zones/microm(3) of tissue), and the number of synapses/neuron (Syn/Neur) were calculated. In FC, significant differences of Nv and Sv due to age were not revealed, while the S value was significantly increased in the aged animals. In TC, Sv did not change in relation to age, whereas Nv was significantly decreased and S significantly increased in aged monkeys. A percent distribution of S showed that the fraction of enlarged synapses (>0.20 microm(2)) was higher in TC than in FC, regardless of the age of the animals (21.3% versus 16.9% in adult and 33.9% versus 26.0% in aged monkeys, respectively). In aged animals, Syn/Neur was not significantly decreased in TC and not significantly increased in FC (4.4%). The above morphometric parameters account for the ongoing rearrangements of synaptic ultrastructure, reacting to the environmental stimuli. Our findings provide evidence of an age-related decline of synaptic plasticity in the brain of aged monkeys that is statistically significant in TC. According to current literature data on synaptic structural dynamics, this decay may represent an early and subtle alteration able to trigger the development of senile plaques and neurodegenerative events. PMID- 17405925 TI - Preservation of mitochondrial volume homeostasis at the early stages of age related synaptic deterioration. AB - A morphometric study on synaptic mitochondria was performed in the frontal (FC) and temporal (TC) cortex of adult and aged monkeys to seek ultrastructural alterations due to age. The overall volume covered by mitochondria (volume density: Vv), the number of mitochondria/microm(3) of tissue (numeric density: Nv), the average mitochondrial size (average volume: V), and the average mitochondrial shape (average length: Fmax) were calculated. Either in FC and TC, no significant age-related differences were revealed for any of the above mentioned morphometric parameters. Namely, in FC of aged monkeys, a decrease of Vv (2%) and Nv (6%) was observed, whereas V and Fmax were increased by 5% and 2%, respectively. In TC of aged animals, both Vv and Nv increased by 7%, V decreased by 2%, and Fmax increased by 1%. The above morphometric parameters account for changes in single aspects of mitochondrial ultrastructure; nonetheless, when considered together per experimental group, they provide information regarding the structural rearrangements occurring on discrete populations of organelles. Considering these assumptions, the present findings document a preservation of the mitochondrial volume homeostasis in the brain of aged monkeys. Because our data from a previous investigation on the same animals showed early signs of synaptic deterioration in FC and TC during aging, this seems to be in contrast with the results of the present study. However, the clear age-related preservation of the mitochondrial potential for structural dynamics may be interpreted as a reactive response to early signs of synaptic deterioration. PMID- 17405926 TI - Immunohistochemical evidence for impaired neuregulin-1 signaling in the prefrontal cortex in schizophrenia and in unipolar depression. AB - In the central nervous system (CNS), neuregulin-1 (NRG-1) proteins function in neuronal migration, differentiation, and survival of oligodendrocytes. The NRG-1 gene codes for at least 15 different isoforms, which may be classified on the basis of their molecular structure. At least two different haplotypes of the NRG 1 gene may be associated with schizophrenia. An abnormal expression pattern of NRG-1 mRNA was found in the prefrontal cortex of schizophrenic patients in comparison to controls. We here show that the NRG-1alpha isoform is significantly reduced in white matter of the prefrontal cortex in schizophrenia but not in affective disorder. In the prefrontal gray matter, the density of NRG-1alpha expressing neurons was reduced in individuals with schizophrenia and in unipolar patients. We studied brains of 22 schizophrenics, 12 patients with affective disorders (7 unipolar and 5 bipolar), and 22 matched controls. NRG-1alpha immunoreactive material was detected with a polyclonal antiserum against the synthetic peptide from alpha-type EGF-like domain of human NRG. The demonstrated decreased number of NRG-1 immunoreactive neurons in the brains of schizophrenics and patients with unipolar depression points to an important role of this NRG 1alpha splice variant in neuropsychiatric disorders. Reduced NRG-1alpha protein concentrations were found in brains of schizophrenics after Western blot analysis. The diminished expression of NRG-1alpha strongly supports an early neurodevelopmental component to schizophrenia. PMID- 17405927 TI - Dysregulation of GABAergic neurotransmission in mood disorders: a postmortem study. AB - Alterations of GABAergic neurotransmission are assumed to play a crucial role in the pathophysiology of mood disorders. Gamma-aminobutyric acid (GABA) acts via binding to A and B receptors, whereas the B receptor is G protein-coupled. Glutamic acid decarboxylase (GAD) is the key enzyme of GABA synthesis. Immunohistochemical staining of GAD 65/67-immunoreactive neurons was performed in dorsolateral prefrontal cortex, orbitofrontal cortex, anterior cingulate cortex, superior temporal cortex, hippocampus formation, and mediodorsal thalamus with consecutive determination of neuronal density in 20 brains of patients with mood disorders (P) and 19 controls (C). In the patients' group were 11 patients with bipolar disorder (BD) and 9 patients with major depressive disorder (MDD). The data were tested statistically using analysis of variance (ANOVA) and post hoc Tukey tests. ANOVA revealed significant differences among the groups (C, BD, MDD) in dorsolateral prefrontal cortex, orbitofrontal cortex, superior temporal cortex, and hippocampus. Post hoc tests demonstrated higher neuronal densities in unipolar patients compared with bipolar patients and controls in dorsolateral prefrontal cortex, superior temporal cortex, and hippocampus. In the orbitofrontal cortex, a higher neuronal density was found in bipolar and unipolar patients compared with controls. In mood disorder patients, dose equivalents of antidepressants given prior to death correlated positively with the neuronal density in superior temporal cortex and hippocampus. The current data on GAD 65/67 point to a dysregulation of the GABAergic system in mood disorders. Possibly, existing deficits of GABAergic neurotransmission will be compensated or overcompensated by antidepressants. Additionally, albeit speculative, an imbalance between GABA production and transport might be of relevance. PMID- 17405928 TI - Release of beta-amyloid from high-density platelets: implications for Alzheimer's disease pathology. AB - The main component of Alzheimer's disease (AD) senile plaques in the brain is amyloid-beta peptide (Abeta), a proteolytic fragment of the amyloid precursor protein (APP). Platelets contain both APP and Abeta and much evidence suggests that these cells may represent a useful tool to study both amyloidogenic and nonamyloidogenic pathways of APP processing. It has been demonstrated that platelets activated by physiological agonists, such as thrombin and collagen, specifically secrete Abeta ending at residue 40. To verify whether APP beta processing could be observed also in an in vitro system of highly concentrated platelets, we measured the Abeta released in the incubation media of 5 x 10(9) platelets/mL by enzyme-linked immunosorbent assay (ELISA). The activation status of platelets was investigated by ultrastructural analysis. We found that Abeta(40) levels were significantly higher in incubation media of 5 x 10(9)/mL platelets in comparison with 10(8)/mL platelets (normalized values), while Abeta(42) levels were not affected by cell density. The ultrastructural analysis showed platelets at different phases of activation: some platelets were at earlier stage, characterized by granule swelling and dilution, others had granules concentrated in a compact mass in the cell centers within constricted rings of circumferential microtubules (later stage). Normally concentrated cells had the characteristic morphology of resting platelets. Our data suggest that high-density platelets undergo activation likely by increased frequency of platelet-platelet collisions. This, in turn, determines the activation of APP beta-processing with consequent release of Abeta(40). Investigating the biochemical pathways triggering Abeta secretion in platelets might provide important information for developing tools to modulate this phenomenon in AD brains. PMID- 17405929 TI - The role of selenite on microglial migration. AB - Oxidative brain damage, such as excitotoxicity and stroke, leads to primary neuronal destruction. The primary damage is further potentiated by macrophages and microglial cells, which are attracted and invade into the zone of damage resulting in secondary neuronal death. Since the essential trace element selenium has anti-inflammatory properties, we analyzed the effects of selenium on these inflammatory cells. Here, we show that the essential trace element selenium abrogates the stress-induced migration of microglial cells. Thus, the antimigratory effects of selenium may attenuate the secondary cell death cascade by preventing microglial invasion. PMID- 17405930 TI - Altered subcellular distribution of the Alzheimer's amyloid precursor protein under stress conditions. AB - Altered metabolism of the Alzheimer's amyloid precursor protein (APP) appears to be a key event in the pathogenesis of Alzheimer's disease (AD), and both altered phosphorylation and oxidative stress appear to affect the production of the toxic Abeta fragment. Our results show that altered processing of APP was observed under conditions of stress induced by sodium azide in the presence of 2-deoxy-D glucose (2DG). As previously reported, the production of the secreted fragment of APP (sAPP) was inhibited. Using APP-GFP fusion proteins, we show that 2DG causes the accumulation/delay of APP in the endoplasmic reticulum (ER)/Golgi (G). The 751 isoform accumulated preferentially in the G, whereas the 695 isoform was blocked preferentially at the ER. This effect was augmented in the presence of sodium azide. APP subcellular distribution was also affected at the plasma membrane. The involvement of protein phosphorylation in APP subcellular localization was reinforced by the effect of drugs, such as phorbol 12-myristate 13-acetate (PMA), since APP was completely depleted from the membrane in the presence of 2DG and PMA. Thus, the hypothesis that APP is processed in a phosphorylation-dependent manner and that this may be of clinical relevance appears to hold true even under stress conditions. Our results provide evidence for a role of protein phosphorylation in APP sorting under stress conditions and contribute to the understanding of the molecular basis of AD. PMID- 17405931 TI - Differential distribution of Alzheimer's amyloid precursor protein family variants in human sperm. AB - The Alzheimer's amyloid precursor protein (APP) is a type I transmembrane glycoprotein with receptor-like characteristics that originates the Abeta peptide by proteolytic processing. Abeta is potentially cytotoxic and the major component of the cerebral amyloid plaques in Alzheimer's disease (AD) patients. APP is known to be ubiquitously expressed in mammalian cells, with a broad tissue distribution, and Abeta deposition has been reported to occur also in many cells outside the nervous system. Although many putative functions have been suggested for APP, its precise physiological role remains to be elucidated. As several results point to a role of chronic inflammation in AD pathogenesis and suggest that AD might be a systemic disorder, the importance of APP function in non neuronal cells/tissues has gained increased relevance. Previous studies have shown that amyloid precursor-like protein 2 (APLP2) is highly expressed in testis and sperm, but failed to unambiguously prove the presence of APP itself in mammalian sperm. The use of a battery of available antibodies that detect APP specific epitopes or epitopes shared with other APP family members, revealed quite distinct distributions in human sperm. Our results are consistent with previous observations of APLP2 in sperm and unequivocally demonstrate the presence of APP itself in human sperm, thus suggesting a putative role for this important protein in sperm function. PMID- 17405932 TI - The effect of repeated physical exercise on hippocampus and brain cortex in stressed rats. AB - Sensitivity of target cells to glucocorticoids is regulated by the expression of intracellular glucocorticoid receptor (GR), which mediates the effects of glucocorticoids. The level of GR and of its nuclear transporter protein 70 (Hsp70) were followed in hippocampus and brain cortex of adult Wistar rat males exposed to acute (immobilization, cold) and chronic (social isolation, isolation, and 15 min daily swimming) stress or their combinations. Changes in plasma levels of adenocorticotropic hormone and corticosterone were also studied. A significant decrease in cytosol GR and Hsp70 was observed after acute stress. Opposite to that, chronic stress led to negligible changes in both cytosol GR and Hsp70 levels. Isolation, as chronic psychosocial stressor, caused reduced responsiveness to novel acute stressors, judged by the cytosol GR and Hsp70 levels. This was not observed if chronic isolation was combined with 15 min daily swimming prior to acute exposure to immobilization. The data suggest that repeated physical exercise may, at least in some cases, diminish detrimental effects of chronic social isolation on limbic-hypothalamic-pituitary adrenocortical axis, as judged by the levels of GR and Hsp70 in the Wistar rat brain. PMID- 17405933 TI - Prion protein aggregation and neurotoxicity in cortical neurons. AB - Prion diseases are degenerative disorders of the central nervous system characterized by cerebral protein aggregation and deposition. A cellular glycoprotein, PrP(C) is converted in an altered isoform, PrP(Sc), that accumulates in the brain, and is believed to be responsible for the neuronal loss observed in prion diseases. The synthetic peptide PrP(106-126) shares many characteristics with PrP(Sc) and is largely used to explore the toxic mechanisms underlying prion diseases. In this article we analyzed the neurotoxic effects of PrP(106-126) in primary rat brain cortical neurons, correlating these results with the presence of amyloid plaques in cultures. Incubation of cells with PrP(106-126), 25 muM, for 2 days did not significantly decrease neuronal viability, although we have observed an increase of basal intracellular calcium levels, reactive oxygen species (ROS) formation, and lipid peroxidation. The presence of congophylic and thioflavin S-amyloid-positive plaques in cortical cultures was only observed after a 5-day-treatment period, correlating with a significant decrease of neuronal viability, as assessed by 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactate dehydrogenase (LDH) leakage. The data obtained support the idea that PrP(106-126) aggregates in vitro and that the aggregation state is important for its neurotoxicity but also suggest that this synthetic peptide, even when is not aggregated in vitro, can compromise cell homeostasis. PMID- 17405934 TI - Intensive remodeling of Purkinje cell spines after climbing fibers deafferentation does not involve MAPK and Akt activation. AB - Subtotal lesion of the inferior olive (IO) achieved by treating experimental animals with 3-acetylpyridine (3AP) induces partial Purkinje cells (PCs) deafferentation that leads to PC hyperactivity and new spine formation. Coincidentally, the olivary terminals belonging to the few survived olivary neurons undergo an extensive collateral sprouting resulting in reinnervation of the neighboring denervated PCs. We obtained chemical deafferentation of PCs in adult rats (body weight, 120-170 g; age, 35-40 days) by a single intraperitoneal injection of 3AP (65 mg/kg body weight), and as early as 3 days after 3AP treatment, important morphological changes could be observed on PCs. Mitogen activated protein kinase (MAPK) cascades and more specifically extracellular signal-regulated kinases 1/2 (ERK1/2) play a critical role in the signaling events underlying synaptic plasticity. For instance, long-term depression (LTD) in the adult hippocampus and long-term potentiation (LTP) in cerebellum both involve ERK activation. Since PCs deprived of their climbing fibers (CFs) afferents initiate an intensive remodeling of the spines and rapid recall of the remaining CFs, it prompted us to see whether the observed phenomena correlated with MAPK and Akt activation. Immunohistochemistry and Western blotting were done at various time points after 3AP application (from 24 h to 6 days), as the exact dynamics of CF loss is not precisely known. As judged by Western blotting, there was no increase of activated ERK in the cerebellum. However, immunohistochemistry revealed increased ERK phosphorylation in the "pinceaux" of basket cells in 3AP animals. Similarly, stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK), p38 MAPK, and Akt activation were also studied by means of Western blotting and immunohistochemistry. Upon 3AP treatment no changes in phosphorylation status could be seen in the different kinases subjected to analysis. Our results suggest that activation of MAPK and Akt cascades is not essential in this model of neuronal plasticity. PMID- 17405935 TI - Immunomorphological analysis of RAGE receptor expression and NF-kappaB activation in tissue samples from normal and degenerated intervertebral discs of various ages. AB - We immunohistochemically investigated the pattern of RAGE expression and NFkappaB translocation into the nucleus in 43 complete cross-sections of human lumbar intervertebral discs (neonatal-85 years) and compared this with the carboxymethyl lysine (CML) modification of proteins as a marker for oxidative stress. No significant expression of RAGE, no obvious activation of NF-kappaB, and no deposition of CML-modified proteins were seen in fetal, juvenile, and young adolescent discs (until age of 13 years). In adults, 25-50% of nucleus pulposus cells were labeled for RAGE and activated NF-kappaB, which correlated well with the occurrence and extent of CML staining in the pericellular matrix. In the annulus fibrosus significantly lower values were seen than in the nucleus pulposus. In consequence, we provide evidence for activation of the NF-kappaB system in intervertebral discs in vivo, which correlates with accumulated oxidative stress and increases in age and disc degeneration. Oxidative stress (as monitored by CML modifications) may lead to RAGE activation and NF-kappaB translocation. PMID- 17405936 TI - Amyloid precursor protein and Presenilin 1 interaction studied by FRET in human H4 cells. AB - The mayor pathologic hallmarks of Alzheimer's disease (AD) are senile plaque and neurofibrillary tangles. Senile plaque are primarily made up of deposits of amyloid-beta protein, a proteolytic product derived from the amyloid precursor protein (APP). APP is a transmembrane protein detected into the endoplasmic reticulum, in the Golgi apparatus, at the cell surface, recycled by endocytosis to endosomes, whose physiological function is unclear. Presenilins (PS), are a component of gamma-secretase complex that cleave alpha-CTFs (carboxy-terminal fragment), or beta-CTFs, leaving 40 or 42 amino acids amyloid-beta peptides and 58 or 56 amino acids intracellular domains (AICD). Where the amyloid-beta peptides is generated is not clear. The study of APP-PS interaction in specific cell compartments provides a good opportunity to light upon the molecular mechanisms regulating the activity of the "gamma-secretase complex," and where beta-amyloid is generated. In our study we used a biophysical assay of protein proximity: fluorescence resonance energy transfer (FRET), that can provide information about molecular interactions when two proteins are in the close proximity (<10 nm), to examine the subcellular localization and interaction between APP and PS1 in human neuroglioma cells (H4). Confocal microscopic analysis reveals extensive colocalization in different cells' compartment, and centrosomal or microtubule organizing center (MTOC) localization of APP and PS1, but not necessarily a close molecular interaction. We used FRET to determine if APP and PS1 interact at the cell centrosome. FRET data suggest a close interaction between APP and PS1 in subcellular compartments and at the centrosome of H4 cells. Using this approach we show that APP and PS1 are closely associated in the centrosomes of the H4 cell. PMID- 17405937 TI - Amino-terminally truncated prion protein PrP90-231 induces microglial activation in vitro. AB - The conversion of the prion protein (PrP) into a protease-resistant isoform (PrP(Res)) is considered the pathogenic event responsible for prion encephalopathies. Microglia activation accompanies PrP(Res) deposition representing an early event in the progression of these diseases. It is now believed that microglial cells play a worsening, if not causative, role in prion induced neuronal death, through the release of proinflammatory and neurotoxic molecules. Indeed, in vitro observations have demonstrated that PrP(Res) and the synthetic prion fragment PrP106-126 induce neuronal death by activating microglial to migrate in the lesion area and secrete cytokines. Recently, we and others have demonstrated that the recombinant peptide, corresponding to the protease-resistant portion of PrP encompassing the amino acids 90-231 (PrP90 231), when beta-structured, is toxic for neuronal cells, in vitro. Here we report that PrP90-231 induces activation of N9 microglial cells, characterized by cell proliferation arrest and increased secretion of different cytokines (RANTES, GCSF, and IL-12). Moreover, the treatment of N9 cells with PrP90-231 elicited inducible nitric oxide synthase (i-NOS) expression, nitric oxide release, and a delayed (15 min to 1 h of treatment) extracellular signal-regulated kinases 1/2 (ERK1/2) phosphorylation/activation. Although ERK1/2 is known to regulate proliferative and differentiative events, we show that its blockade, using the specific MEK inhibitor PD98059, did not prevent PrP90-231-induced inhibition of N9 cell proliferation. To our knowledge, this is the first evidence that a recombinant PrP(Res)-like peptide elicits microglial activation in vitro, thus representing a potentially important tool to develop possible therapeutic strategies to target prion-induced brain inflammation. PMID- 17405938 TI - Activation and endocytic internalization of melanocortin 3 receptor in neuronal cells. AB - Melanocortins play a central role in autonomic modulation of metabolism by acting through a family of highly homologous G protein-coupled receptors. Studies with gene knockout mice have implicated neural melanocortin receptors, MC3R and MC4R, in the etiology of obesity, insulin resistance, and salt-sensitive hypertension. In an attempt to better understand the mechanisms of function of these receptors, we expressed MC3R and MC4R in neuronal cells and demonstrated their co localization to several membrane regions. We now show that in cultured neuronal cells, MC3R localizes to lipid rafts and undergoes endocytic internalization upon activation by gamma-MSH through a protein kinase-sensitive pathway. The appearance of the internalized receptor in lysosomes suggests that it is subsequently degraded. The expression of protein kinase A regulatory subunits and of c-Jun and c-Fos was analyzed by either immunoblotting or real-time PCR. No discernable changes were observed in the expression levels of these protein kinase A and protein kinase C responsive genes. Immunohistochemical studies showed a robust expression of MC3R protein in brain nuclei with relevance to cardiovascular function and fluid homeostasis further supporting the notion that the physiological effects of melanocortins on the cardiovascular system arise from effects on the central nervous system. PMID- 17405939 TI - Biomarkers for cystic fibrosis: are we progressing? PMID- 17405940 TI - Hold the front page: smoking bans good for (most) workers' health. PMID- 17405941 TI - Update in cystic fibrosis 2006. PMID- 17405942 TI - Update in environmental and occupational medicine 2006. PMID- 17405943 TI - Antituberculosis drugs and hepatotoxicity. PMID- 17405944 TI - Complications at birth and subsequent wheeze: risk of attrition bias. PMID- 17405945 TI - Toward optimizing treatment for Mycobacterium avium complex lung disease. PMID- 17405946 TI - A new era in protein quantification in clinical laboratories: application of liquid chromatography-tandem mass spectrometry. PMID- 17405947 TI - Considerably reduced centrifugation time without increased hemolysis: evaluation of the new BD Vacutainer SSTTMII Advance. PMID- 17405948 TI - Liposomal amphotericin B interferes with the phosphorus assay on the Synchron LX 20 analyzer. PMID- 17405949 TI - Epigenetic analysis of RASSF1A gene in cell-free DNA in amniotic fluid. PMID- 17405950 TI - Comparison of calcitonin determinations by polyclonal and monoclonal IRMAs. PMID- 17405951 TI - Total or neonatal bilirubin assays in the Vitros 5,1 FS: hemoglobin interference, hemolysis, icterus index. PMID- 17405952 TI - Very high inhibin A concentration attributed to heterophilic antibody interference. PMID- 17405953 TI - Plasma total coenzyme Q9 (CoQ9) in the New Zealand population: reference interval and biological variation. PMID- 17405954 TI - Analytical variation in plasma renin activity: implications for the screening of primary aldosteronism. PMID- 17405955 TI - The impact of simultaneous measurement of testosterone and androstenedione in women with suspected androgen excess. PMID- 17405956 TI - The cover. Umbrellas in the Rain. PMID- 17405957 TI - A piece of my mind. A question of faith. PMID- 17405958 TI - Brain scans, genes provide addiction clues. PMID- 17405959 TI - A change of heart guidelines for women. PMID- 17405964 TI - Proton pump inhibitor therapy and hip fracture risk. PMID- 17405965 TI - Medicare's Hospital Compare performance measures and mortality rates. PMID- 17405966 TI - Medicare's Hospital Compare performance measures and mortality rates. PMID- 17405967 TI - High-altitude pulmonary edema and patent foramen ovale. PMID- 17405968 TI - High-altitude pulmonary edema and patent foramen ovale. PMID- 17405969 TI - Emergence of influenza B viruses with reduced sensitivity to neuraminidase inhibitors. AB - CONTEXT: Very little is known about the frequency of generation and transmissibility of influenza B viruses with reduced sensitivity to neuraminidase inhibitors. Furthermore, transmission of resistant virus, whether influenza A or B, has not been recognized to date. OBJECTIVE: To assess the prevalence and transmissibility of influenza B viruses with reduced sensitivity to neuraminidase inhibitors. DESIGN, SETTING, AND PATIENTS: Investigation of the neuraminidase inhibitor sensitivity of influenza B isolates from 74 children before and after oseltamivir therapy and from 348 untreated patients with influenza (including 66 adults) seen at 4 community hospitals in Japan during the 2004-2005 influenza season. Four hundred twenty-two viruses from untreated patients and 74 samples from patients after oseltamivir therapy were analyzed. MAIN OUTCOME MEASURE: Sialidase inhibition assay was used to test the drug sensitivities of influenza B viruses. The neuraminidase and hemagglutinin genes of viruses showing reduced sensitivity to neuraminidase inhibitors were sequenced to identify mutations that have the potential to confer reduced sensitivity to these drugs. RESULTS: In 1 (1.4%) of the 74 children who had received oseltamivir, we identified a variant with reduced drug sensitivity possessing a Gly402Ser neuraminidase substitution. We also identified variants with reduced sensitivity carrying an Asp198Asn, Ile222Thr, or Ser250Gly mutation in 7 (1.7%) of the 422 viruses from untreated patients. Review of the clinical and viral genetic information available on these 7 patients indicated that 4 were likely infected in a community setting, while the remaining 3 were probably infected through contact with siblings shedding the mutant viruses. CONCLUSIONS: In this population, influenza B viruses with reduced sensitivity to neuraminidase inhibitors do not arise as frequently as resistant influenza A viruses. However, they appear to be transmitted within communities and families, requiring continued close monitoring. PMID- 17405970 TI - Sumatriptan-naproxen for acute treatment of migraine: a randomized trial. AB - CONTEXT: Multiple pathogenic mechanisms may be involved in generating the migraine symptom complex, and multimechanism-targeted therapy may confer advantages over monotherapy. OBJECTIVE: To evaluate the efficacy and safety of a fixed-dose tablet containing sumatriptan succinate and naproxen sodium relative to efficacy and safety of each monotherapy and placebo for the acute treatment of migraine. DESIGN, SETTING, AND PARTICIPANTS: Two replicate, randomized, double blind, single-attack, parallel-group studies conducted among 1461 (study 1) and 1495 (study 2) patients at 118 US clinical centers who were diagnosed as having migraine and received study treatment for a moderate or severe migraine attack. INTERVENTIONS: Patients were randomized in a 1:1:1:1 ratio to receive a single tablet containing sumatriptan, 85 mg, and naproxen sodium, 500 mg; sumatriptan, 85 mg (monotherapy); naproxen sodium, 500 mg (monotherapy); or placebo, to be used after onset of a migraine with moderate to severe pain. MAIN OUTCOME MEASURES: Primary outcome measures included the percentages of patients with headache relief 2 hours after dosing, absence of photophobia, absence of phonophobia, and absence of nausea for the comparison between sumatriptan naproxen sodium and placebo, and the percentages of patients with sustained pain free response for the comparison between sumatriptan-naproxen sodium and each monotherapy. RESULTS: Sumatriptan-naproxen sodium was more effective than placebo for headache relief at 2 hours after dosing (study 1, 65% vs 28%; P<.001 and study 2, 57% vs 29%; P<.001), absence of photophobia at 2 hours (58% vs 26%; P<.001 and 50% vs 32%; P<.001), and absence of phonophobia at 2 hours (61% vs 38%; P<.001 and 56% vs 34%; P<.001). The absence of nausea 2 hours after dosing was higher with sumatriptan-naproxen sodium than placebo in study 1 (71% vs 65%; P = .007), but in study 2 rates of absence of nausea did not differ between sumatriptan-naproxen sodium and placebo (65% vs 64%; P = .71). For 2- to 24-hour sustained pain-free response, sumatriptan-naproxen sodium was superior at P<.01 (25% and 23% in studies 1 and 2, respectively) to sumatriptan monotherapy (16% and 14% in studies 1 and 2), naproxen sodium monotherapy (10% and 10% in studies 1 and 2), and placebo (8% and 7% in studies 1 and 2). The incidence of adverse events was similar between sumatriptan-naproxen sodium and sumatriptan monotherapy. CONCLUSION: Sumatriptan, 85 mg, plus naproxen sodium, 500 mg, as a single tablet for acute treatment of migraine resulted in more favorable clinical benefits compared with either monotherapy, with an acceptable and well-tolerated adverse effect profile. TRIAL REGISTRATION: clinicaltrials.gov Identifiers: NCT00434083 (study 1); NCT00433732 (study 2). PMID- 17405971 TI - Statin use and sepsis events [corrected] in patients with chronic kidney disease. AB - CONTEXT: Patients with chronic kidney disease are at high risk for sepsis and sepsis-related mortality. OBJECTIVE: To assess whether statin use is associated with a reduction in sepsis events [corrected] in dialysis patients. DESIGN, SETTING, AND PATIENTS: National prospective cohort study that enrolled 1041 incident dialysis patients at 81 US not-for-profit outpatient dialysis clinics from October 1995 to June 1998, with follow-up to January 2005. Statin use was determined by medical record review. Rates of sepsis events [corrected] between statin users and control patients were compared using multivariate regression models, with adjustment for potential confounders in the overall cohort and in a subcohort in which control patients were matched to statin users according to their likelihood (propensity) to have been prescribed a statin. MAIN OUTCOME MEASURE: Sepsis events [corrected] were determined through hospital records from the United States Renal Data System (mean follow-up, 3.4 years). RESULTS: There were 303 sepsis events [corrected] Rates of sepsis events [corrected] were significantly lower in patients receiving statins (crude incidence rate, 41/1000 patient-years) than in those not receiving statins (crude incidence rate, 110/1000 patient-years) (P<.001). With adjustment for demographics and dialysis modality, statin users were substantially less likely to be subsequently hospitalized for sepsis (incidence rate ratio, 0.41; 95% confidence interval [CI], 0.25-0.68). Further adjustment for comorbidities and laboratory values continued to show this protective association (incidence rate ratio, 0.38; 95% CI, 0.21-0.67). In the propensity-matched subcohort, statin use was even more protective (incidence rate ratio, 0.24; 95% CI, 0.11-0.49). CONCLUSIONS: Use of statins was strongly and independently associated with a reduction in the risk of sepsis events [corrected] in patients who had chronic kidney disease and were receiving dialysis. Randomized trials of statins in patients with chronic kidney disease should examine the prevention of sepsis as a potentially important benefit. PMID- 17405972 TI - Postmenopausal hormone therapy and risk of cardiovascular disease by age and years since menopause. AB - CONTEXT: The timing of initiation of hormone therapy may influence its effect on cardiovascular disease. OBJECTIVE: To explore whether the effects of hormone therapy on risk of cardiovascular disease vary by age or years since menopause began. DESIGN, SETTING, AND PARTICIPANTS: Secondary analysis of the Women's Health Initiative (WHI) randomized controlled trials of hormone therapy in which 10,739 postmenopausal women who had undergone a hysterectomy were randomized to conjugated equine estrogens (CEE) or placebo and 16,608 postmenopausal women who had not had a hysterectomy were randomized to CEE plus medroxyprogesterone acetate (CEE + MPA) or placebo. Women aged 50 to 79 years were recruited to the study from 40 US clinical centers between September 1993 and October 1998. MAIN OUTCOME MEASURES: Statistical test for trend of the effect of hormone therapy on coronary heart disease (CHD) and stroke across categories of age and years since menopause in the combined trials. RESULTS: In the combined trials, there were 396 cases of CHD and 327 cases of stroke in the hormone therapy group vs 370 [corrected] cases of CHD and 239 cases of stroke in the placebo group. For women with less than 10 years since menopause began, the hazard ratio (HR) for CHD was 0.76 (95% confidence interval [CI], 0.50-1.16); 10 to 19 years, 1.10 (95% CI, 0.84-1.45); and 20 or more years, 1.28 (95% CI, 1.03-1.58) (P for trend = .02). The estimated absolute excess risk for CHD for women within 10 years of menopause was -6 per 10,000 person-years; for women 10 to 19 years since menopause began, 4 per 10,000 person-years; and for women 20 or more years from menopause onset, 17 per 10,000 person-years. For the age group of 50 to 59 years, the HR for CHD was 0.93 (95% CI, 0.65-1.33) and the absolute excess risk was -2 per 10,000 person years; 60 to 69 years, 0.98 (95% CI, 0.79-1.21) and -1 per 10,000 person-years; and 70 to 79 years, 1.26 (95% CI, 1.00-1.59) and 19 per 10,000 person-years (P for trend = .16). Hormone therapy increased the risk of stroke (HR, 1.32; 95% CI, 1.12-1.56). Risk did not vary significantly by age or time since menopause. There was a nonsignificant tendency for the effects of hormone therapy on total mortality to be more favorable in younger than older women (HR of 0.70 for 50-59 years; 1.05 for 60-69 years, and 1.14 for 70-79 years; P for trend = .06). CONCLUSIONS: Women who initiated hormone therapy closer to menopause tended to have reduced CHD risk compared with the increase in CHD risk among women more distant from menopause, but this trend test did not meet our criterion for statistical significance. A similar nonsignificant trend was observed for total mortality but the risk of stroke was elevated regardless of years since menopause. These data should be considered in regard to the short-term treatment of menopausal symptoms. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00000611. PMID- 17405973 TI - Does this adult patient have septic arthritis? AB - CONTEXT: In patients who present with an acutely painful and swollen joint, prompt identification and treatment of septic arthritis can substantially reduce morbidity and mortality. OBJECTIVE: To review the accuracy and precision of the clinical evaluation for the diagnosis of nongonococcal bacterial arthritis. DATA SOURCES: Structured PubMed and EMBASE searches (1966 through January 2007), limited to human, English-language articles and using the following Medical Subject Headings terms: arthritis, infectious, physical examination, medical history taking, diagnostic tests, and sensitivity and specificity. STUDY SELECTION: Studies were included if they contained original data on the accuracy or precision of historical items, physical examination, serum, or synovial fluid laboratory data for diagnosing septic arthritis. DATA EXTRACTION: Three authors independently abstracted data from the included studies. DATA SYNTHESIS: Fourteen studies involving 6242 patients, of whom 653 met the gold standard for the diagnosis of septic arthritis, satisfied all inclusion criteria. Two studies examined risk factors and found that age, diabetes mellitus, rheumatoid arthritis, joint surgery, hip or knee prosthesis, skin infection, and human immunodeficiency virus type 1 infection significantly increase the probability of septic arthritis. Joint pain (sensitivity, 85%; 95% confidence interval [CI], 78% 90%), a history of joint swelling (sensitivity, 78%; 95% CI, 71%-85%), and fever (sensitivity, 57%; 95% CI, 52%-62%) are the only findings that occur in more than 50% of patients. Sweats (sensitivity, 27%; 95% CI, 20%-34%) and rigors (sensitivity, 19%; 95% CI, 15%-24%) are less common findings in septic arthritis. Of all laboratory findings readily available to the clinician, the 2 most powerful were the synovial fluid white blood cell (WBC) count and percentage of polymorphonuclear cells from arthrocentesis. The summary likelihood ratio (LR) increased as the synovial fluid WBC count increased (for counts <25,000/microL: LR, 0.32; 95% CI, 0.23-0.43; for counts > or =25,000/microL: LR, 2.9; 95% CI, 2.5 3.4; for counts >50,000/microL: LR, 7.7; 95% CI, 5.7-11.0; and for counts >100,000/microL: LR, 28.0; 95% CI, 12.0-66.0). On the same synovial fluid sample, a polymorphonuclear cell count of at least 90% suggests septic arthritis with an LR of 3.4 (95% CI, 2.8-4.2), while a polymorphonuclear cell count of less than 90% lowers the likelihood (LR, 0.34; 95% CI, 0.25-0.47). CONCLUSIONS: Clinical findings identify patients with peripheral, monoarticular arthritis who might have septic arthritis. However, the synovial WBC and percentage of polymorphonuclear cells from arthrocentesis are required to assess the likelihood of septic arthritis before the Gram stain and culture test results are known. PMID- 17405974 TI - Fetomaternal cell trafficking and the stem cell debate: gender matters. PMID- 17405975 TI - News about influenza B drug resistance that cannot be ignored. PMID- 17405976 TI - JAMA patient page. Septic arthritis. PMID- 17405977 TI - Mast-cell sarcoma of the tibia. AB - The mast-cell sarcoma of a bone is described here for the first time. The tumour presented in a 4-year-old boy, with pain, oedema and deformation of his right lower leg. Radiological findings revealed a destructive tumourous mass. Histopathological examination showed the tumour to be composed of large, atypical cells, with hyperchromatic oval and polygonal nuclei. The cytoplasm around them was eosinophilic with many basophilic and toluidine-blue-positive granules. These atypical mast cells were positive for chloroacetate esterase, c-kit, tryptase and negative for myeloperoxidase. The primary disease quickly progressed to mast-cell leukaemia, and despite intensive chemotherapy the patient died 18 months after first symptoms. PMID- 17405978 TI - Papillary renal cell carcinoma within a renal oncocytoma: case report of an incidental finding of a tumour within a tumour. AB - The most common renal tumours are clear cell, papillary, chromophobe and collecting duct renal cell carcinomas (RCCs), and benign oncocytomas and angiomyolipomas. Tumours with hybrid features between some of these entities have been recognised; in particular, tumours with features of both chromophobe RCC and oncocytoma. Case reports describing one distinct type of primary renal tumour actually within another are very rare. The incidental finding of a papillary RCC located in an oncocytoma in a nephrectomy specimen from a 75-year-old man is described. Morphological criteria for each tumour type were completely satisfied and fluorescence in situ hybridisation detected the expected number of copies of chromosome 7 in the cells of each tumour type. The cells in the papillary tumour contained three copies, whereas the oncocytoma cells contained only two per nucleus. To our knowledge, this is the first report of a papillary RCC being identified within an oncocytoma. PMID- 17405979 TI - Metastatic retroperitoneal epithelioid angiomyolipoma. AB - Angiomyolipoma (AML) is a rare benign neoplasm that usually arises in the kidneys, but may rarely originate in sites such as the retroperitoneum, liver and bone. It is characterised by an intimate admixture of blood vessels, smooth muscle and fat. This multiphasic composition allows for its recognition on ultrasound and CT scan examination. Metastases are exceedingly uncommon, and only one other case of metastatic extrarenal AML exists in the literature. Histological variants of AML such as epithelioid angiomyolipoma (EAML) are considered to be locally aggressive. A unique case of an EAML of the retroperitoneum that metastasised to the liver and bone in an 80-year-old woman is described. PMID- 17405980 TI - Infection-associated haemophagocytic syndrome associated with recurrent acute myeloid leukaemia/myelodysplastic syndrome: an autopsy case. PMID- 17405981 TI - Kikuchi's disease displaying a t(2:16) chromosomal translocation. AB - Kikuchi's disease is a rare self-limiting lymphoproliferative condition of unknown aetiology, characterised by acute or subacute necrotising lymphadenitis. It is a benign condition that can mimic malignant lymphoma. In this report, a case of Kikuchi's disease associated with a chromosomal abnormality is described. This is the first report in the literature of such a case and it highlights an important learning point; benign lymphoproliferative conditions can be associated with chromosomal abnormalities that are more typically associated with malignant lymphoproliferative conditions such as malignant lymphoma. The report illustrates the necessity for interpreting cytogenetic data in the relevant clinical and histopathological context in a multidisciplinary setting to avoid misdiagnosis and inappropriate treatment. PMID- 17405982 TI - Pseudo-pseudohypercalcaemia, apparent primary hyperparathyroidism and Waldenstrom's macroglobulinaemia. AB - An apparent primary hyperparathyroidism was reported due to pseudo pseudohypercalcaemia in a 74-year-old man with Waldenstrom's macroglobulinaemia. It is important to recognise artificially elevated serum calcium levels so as to avoid erroneous diagnosis, unnecessary investigations and potentially inappropriate treatment. PMID- 17405983 TI - A distinctive vulval fibroma of so-called prepubertal type in a postmenopausal patient. AB - The case of a 65-year-old female patient who has had a large right vulval mass for over 30 years is reported here. The mass was excised, but the hypocellular mesenchymal tumour was difficult to classify histologically. On further specialist assessment, it was found to be identical to the recently recognised distinctive prepubertal vulval fibroma. These are very rarely reported in children, and till now, has never been reported in an adult. PMID- 17405984 TI - Human papilloma virus in squamous carcinoma of the head and neck: a study of cases in south east Scotland. AB - Several studies have found human papillomavirus virus (HPV) in tissue from head and neck squamous cell carcinomas (HNSCCs), although the number of positive cases varies greatly from study to study. The extent and molecular epidemiology of HPV in HNSCC were assessed within cases drawn from southeast Scotland by performing broad-spectrum, real-time HPV polymerase chain reaction (PCR) on DNA extracted from 100 cases of HNSCC in formalin-fixed, paraffin wax-embedded material. All HPV-positive specimens were genotyped and sampled by laser capture microdissection. Pure samples of tumour, and, where possible, dysplastic and normal epithelium were then submitted for further HPV PCR and genotyping to investigate the sensitivity of the technique in small tissue samples. 10 of 100 cases tested positive for HPV, with 8 of these being derived from Waldeyer's ring. HPV DNA was found in adjacent epithelium in two of four cases where this was available. These findings confirm that HPV is likely to be involved in a subset of HNSCC in this population and that successful amplification of HPV nucleic acid is possible even using small amounts of paraffin wax-embedded tissue. PMID- 17405985 TI - Copper:caeruloplasmin ratio. AB - Investigation of copper status can be a diagnostic challenge. The non caeruloplasmin-bound copper (NCC) has deficiencies; accordingly, the copper:caeruloplasmin ratio has been suggested as an alternative index of copper status. A reference interval for this index was derived. In addition to making the interpretation of copper easier, the copper:caeruloplasmin ratio should also enable adjustment for relatively high caeruloplasmin concentrations without recourse to producing gender- and age-derived intervals. The copper:caeruloplasmin ratio has weaknesses similar to those identified for NCC in that immunological methods used for caeruloplasmin can cross react with apocaeruloplasmin and there is no standardised method for caeruloplasmin. Caeruloplasmin assays also have uncertainty from precision, bias and specificity and, accordingly, method-related differences may have a large effect on the copper:caeruloplasmin ratio in a manner similar to the NCC. PMID- 17405986 TI - Primary cardiac post-transplantation lymphoproliferative disorder--T cell type: a case report and review of the literature. PMID- 17405987 TI - A novel rapid and sensitive BRCA1/2 mutation detection method based on pooled DGGE and targeted sequencing. PMID- 17405988 TI - Adjuvant therapy for breast cancer: we are solving the puzzle, but pieces are still missing. PMID- 17405989 TI - Prostate-specific antigen levels, prostate-specific antigen kinetics, and prostate cancer prognosis: a tocsin calling for prospective studies. PMID- 17405990 TI - Users fear that lymphoma drugs will disappear. PMID- 17405991 TI - Zebrafish take the stage in cancer research. PMID- 17405992 TI - Debate over institutional review boards continues as alternative options emerge. PMID- 17405993 TI - Taming a mutinous mutant: an errant receptor becomes a prime cancer target. PMID- 17405994 TI - Stat bite: State spending of tobacco settlement. PMID- 17405995 TI - Polychemotherapy for early breast cancer: results from the international adjuvant breast cancer chemotherapy randomized trial. AB - BACKGROUND: Survival of patients with early-stage breast cancer is improved following treatment with single-modality tamoxifen, ovarian ablation or suppression, or chemotherapy. The Adjuvant Breast Cancer Trials were designed to ascertain any additional benefits of combined treatment. METHODS: The Adjuvant Breast Cancer Chemotherapy Trial was a randomized phase III trial in which patients with early-stage breast cancer who were receiving prolonged (5 years) tamoxifen treatment, with or without ovarian ablation or suppression, were randomly assigned to standard chemotherapy versus none. Trial endpoints included relapse-free and overall survival. Hazard ratios (HRs) were derived from Cox models, and all statistical tests were two-sided. RESULTS: Between 1992 and 2000, 1991 patients between the ages of 26 and 81 years were randomly assigned (987 to chemotherapy, 1004 to no chemotherapy) from 106 UK and 16 non-UK centers. Nine hundred seven (92%) patients received chemotherapy as allocated (87% received cyclophosphamide, methotrexate, and 5-fluorouracil; 11% received anthracycline containing regimens). A total of 244 of the 619 premenopausal patients received elective ovarian ablation or suppression. Chemotherapy improved relapse-free survival (relapse in the chemotherapy group versus no-chemotherapy group, 298 events versus 332 events, HR = 0.86, 95% confidence interval [CI] = 0.73 to 1.01; P = .06) and overall survival (death from any cause in the chemotherapy group versus no-chemotherapy group, 243 events versus 282 events, HR = 0.83, 95% CI = 0.70 to 0.99; P = .03) after adjustment for nodal status, estrogen receptor status, and age. Subgroup analyses showed that the benefit of chemotherapy was greatest in younger women (<50 years) and in particular for premenopausal women not receiving ovarian ablation or suppression. CONCLUSION: Modest yet sustainable benefits for chemoendocrine therapy occur in women with breast cancer. However, the full impact on overall survival may not emerge for several years. PMID- 17405996 TI - Ovarian ablation or suppression in premenopausal early breast cancer: results from the international adjuvant breast cancer ovarian ablation or suppression randomized trial. AB - BACKGROUND: Substantial survival benefits exist for patients with early-stage breast cancer who undergo treatment with single-modality tamoxifen, ovarian ablation or suppression, or chemotherapy. To determine whether additional benefits exist with combined treatment, the Adjuvant Breast Cancer (ABC) Trials were undertaken. METHODS: The ABC Ovarian Ablation or Suppression Trial randomly assigned pre- and perimenopausal patients with early-stage breast cancer who were receiving prolonged (5 years) tamoxifen treatment with or without chemotherapy to ovarian ablation or suppression (by oophorectomy, ovarian irradiation, or treatment with luteinizing hormone-releasing hormone agonist) versus no ovarian ablation or suppression. Trial endpoints included relapse-free and overall survival. Hazard ratios (HRs) were derived from Cox models, and all statistical tests were two-sided. RESULTS: Between 1993 and 2000, 2144 (1063 ovarian ablation or suppression, 1081 no ovarian ablation or suppression) patients were randomly assigned. A total of 942 (89%) received ovarian ablation or suppression as allocated. Overall, no evidence of a benefit for ovarian ablation or suppression was observed for relapse-free survival (relapse in the ovarian ablation/suppression versus no ovarian ablation/suppression group, 290 events versus 306 events, HR = 0.95, 95% confidence interval [CI] = 0.81 to 1.12; P = .56) or overall survival (death from any cause in the ovarian ablation or suppression versus no ovarian ablation/suppression group, 215 events versus 230 events, HR = 0.94, 95% CI = 0.78 to 1.13; P = .44), nor were differences seen after adjustment for age, nodal status, or estrogen receptor (ER) status. CONCLUSION: Overall, no added effect of ovarian ablation or suppression was seen on relapse-free survival or overall survival of premenopausal women who were treated for early-stage breast cancer. However, the role of ovarian ablation or suppression in young (<40 years) women with ER-positive tumors, especially those not receiving chemotherapy, requires further study. PMID- 17405997 TI - Prostate-specific antigen levels as a predictor of lethal prostate cancer. AB - BACKGROUND: Rates of long-term survival among patients with untreated localized prostate cancer are high. To avoid unnecessary treatment, tools are needed to identify the small proportion of patients who are destined to develop lethal prostate cancer. METHODS: To evaluate the accuracy of early changes in prostate specific antigen (PSA) levels as predictors of prostate cancer outcome, we assessed serial measurements of PSA level among 267 men with localized prostate cancer in a Scandinavian cohort of men who were diagnosed between 1989 and 1999 and who were managed by watchful waiting. We then 1) fitted individual regression lines to the PSA values assessed for each patient during the first 2 years of follow-up by using three different models, 2) evaluated early PSA curve characteristics as determinants of the cumulative incidence of lethal prostate cancer and calculated hazard ratios for baseline PSA value and rate of change in PSA level to prostate cancer outcome, and 3) plotted time-dependent receiver operating characteristic (ROC) curves. All P values are two-sided. RESULTS: During complete follow-up for a mean of 8.5 years, 34 patients (13%) died from prostate cancer, and 18 (7%) developed metastases but were still alive at end of follow-up. In a log-linear model, both PSA value at baseline (P = .05) and the rate of PSA change (P<.001) were associated with the development of lethal prostate cancer. In the ROC analysis, however, the accuracy of classifying the disease as either indolent or destined to progress was low, regardless of the cut point chosen for initial PSA level or rate of change in PSA level. CONCLUSIONS: Although baseline PSA value and rate of PSA change are prognostic factors for lethal prostate cancer, they are poor predictors of lethal prostate cancer among patients with localized prostate cancer who are managed by watchful waiting. PMID- 17405998 TI - Noncancer causes of death in survivors of testicular cancer. AB - BACKGROUND: Although modern treatments for testicular cancer are associated with increased survival, the long-term health effects of these treatments are unclear. We conducted a population-based study to quantify the long-term risks of mortality from noncancer causes among men with testicular cancer. METHODS: We identified 38,907 one-year survivors of testicular cancer within 14 population based cancer registries in North America and Europe (from 1943 through 2002). We used data from these registries to calculate standardized mortality ratios (SMRs) for noncancer deaths and to evaluate associations between histology, age at testicular cancer diagnosis, calendar year of diagnosis, and initial treatment and the risk of noncancer mortality. All statistical tests were two-sided. RESULTS: A total of 2942 deaths from all noncancer causes were reported after a median follow-up of 10 years, exceeding the expected number of deaths from all noncancer causes in the general population by 6% (SMR = 1.06, 95% confidence interval [CI] = 1.02 to 1.10); the noncancer standardized mortality ratios did not differ statistically significantly between patients diagnosed before and after 1975, when cisplatin-based chemotherapy came into widespread use. Compared with the general population, testicular cancer survivors had higher mortality from infections (SMR = 1.28, 95% CI = 1.12 to 1.47) and from digestive diseases (SMR = 1.44, 95% CI = 1.26 to 1.64). Mortality from all circulatory diseases was statistically significantly elevated in men diagnosed with testicular cancer before age 35 years (1.23, 95% CI = 1.09 to 1.39) but not in men diagnosed at older ages (SMR = 0.94; 95% CI = 0.89 to 1.00). Men treated with chemotherapy (with or without radiotherapy) in 1975 or later had higher mortality from all noncancer causes (SMR = 1.34, 95% CI = 1.15 to 1.55), all circulatory diseases (SMR = 1.58, 95% CI = 1.25 to 2.01), all infections (SMR = 2.48, 95% CI = 1.70 to 3.50), and all respiratory diseases (SMR = 2.53, 95% CI = 1.26 to 4.53). Testicular cancer patients who were younger than 35 years at diagnosis and were treated with radiotherapy alone in 1975 or later had higher mortality from all circulatory diseases (SMR = 1.70, 95% CI = 1.21 to 2.31) compared with the general population. CONCLUSION: Men who have survived for at least 1 year after being diagnosed with testicular cancer have a slightly higher risk of dying from noncancer causes, including infections, digestive diseases, and circulatory diseases, than the general population. Men treated with chemotherapy in 1975 or later may be at particularly high risk. PMID- 17405999 TI - Secondary chemoprevention of Barrett's esophagus with celecoxib: results of a randomized trial. AB - BACKGROUND: Barrett's esophagus is a premalignant condition that is a risk factor for the development of esophageal adenocarcinoma, a disease whose incidence is rapidly increasing. Because aspirin and other nonsteroidal anti-inflammatory drugs, such as celecoxib, may decrease the risk of developing esophageal cancer, we investigated the effect of long-term administration of celecoxib in patients with Barrett's esophagus with dysplasia. METHODS: Chemoprevention for Barrett's Esophagus Trial (CBET) is a phase IIb multicenter randomized placebo-controlled trial of celecoxib in patients with Barrett's esophagus and low- or high-grade dysplasia. Patients were randomly assigned to treatment with 200 mg of celecoxib or placebo, both administered orally twice daily, and then stratified by grade of dysplasia. The primary outcome was the change from baseline to 48 weeks of treatment in the proportion of biopsy samples with dysplasia between the celecoxib and placebo arms. Secondary and tertiary outcomes included evaluation of changes in histology and expression levels of relevant biomarkers. All statistical tests were two-sided. RESULTS: From April 1, 2000, through June 30, 2003, 222 patients were registered into CBET, and 100 of them with low- or high grade Barrett's dysplasia were randomly assigned to treatment (49 to celecoxib and 51 to placebo). After 48 weeks of treatment, no difference was observed in the median change in the proportion of biopsy samples with dysplasia or cancer between treatment groups in either the low-grade (median change with celecoxib = 0.09, interquartile range [IQR] = -0.32 to 0.14 and with placebo = -0.07, IQR = 0.26 to 0.12; P = .64) or high-grade (median change with celecoxib = 0.12, IQR = 0.31 to 0.55, and with placebo = 0.02, IQR = -0.24 to 0.28; P = .88) stratum. No statistically significant differences in total surface area of the Barrett's esophagus; in prostaglandin levels; in cyclooxygenase-1/2 mRNA levels; or in methylation of tumor suppressor genes p16, adenomatous polyposis coli, and E cadherin were found with celecoxib compared with placebo. CONCLUSIONS: Administration of 200 mg of celecoxib twice daily for 48 weeks of treatment does not appear to prevent progression of Barrett's dysplasia to cancer. PMID- 17406000 TI - Promotion of bladder cancer development and progression by androgen receptor signals. AB - BACKGROUND: Males have a higher incidence of bladder cancer than females, but the reason remains unknown. Unlike prostate cancer, human bladder cancer is not generally considered to be dependent on hormone activity. We investigated the possible involvement of androgens and the androgen receptor (AR) in bladder cancer. METHODS: We used N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) to induce bladder cancer in wild-type male and female mice, with and without castration in males, and in AR knockout (ARKO) male and female mice, with and without dihydrotestosterone (DHT) supplementation in males. We also treated human bladder cancer cell lines, including TCC-SUP and UMUC3, and mouse xenograft models established from these same lines with androgen deprivation therapy (antiandrogen treatment or castration), AR-small-interfering RNA (AR-siRNA), or the anti-AR molecule ASC-J9, which causes selective degradation of the AR. RESULTS: More than 92% of wild-type male and 42% of wild-type female mice treated with BBN eventually developed bladder cancer, whereas none of the male or female ARKO mice did. Treatment with BBN induced bladder cancer in 25% of ARKO mice supplemented with DHT and in 50% of castrated wild-type male mice. Androgen deprivation of AR positive human bladder cancer cells by androgen depletion in vitro or castration in mice and/or by treatment with the antiandrogen flutamide in vitro or in vivo, as well as AR knockdown by AR-siRNA or by ASC-J9, suppressed cell proliferation in vitro and xenograft tumor growth in vivo. CONCLUSIONS: Our findings implicate the involvement of both androgens and the AR in bladder cancer. Targeting AR and androgens may provide novel chemopreventive and therapeutic approaches for bladder cancer. PMID- 17406001 TI - Re: Rising incidence of small renal masses: a need to reassess treatment effect. PMID- 17406003 TI - Re: Age-related lobular involution and risk of breast cancer. PMID- 17406004 TI - Re: Breast cancer gene microarrays pass muster. PMID- 17406006 TI - Using probabilistic corrections to account for abstractor agreement in medical record reviews. AB - The quality of medical record abstracts is often characterized in a reliability substudy. These results usually indicate agreement, but not the extent to which lack of agreement affects associations observed in the complete data. In this study, medical records were reviewed and abstracted for patients diagnosed with stage I or stage II breast cancer between 1990 and 1994 at one of six US Cancer Research Network sites. For a subsample, interrater reliability data were available. The authors calculated conventional hazard ratios and 95% confidence intervals for the association of demographic, tumor, and treatment characteristics with recurrence rate. These conventional estimates of effect were compared with three sets of estimates and 95% simulation intervals that took account of the uncertainty assessed by lack of agreement in the reliability substudy. The rate of recurrence was associated with increasing cancer stage and with treatment modality but not with demographic characteristics. The hazard ratios and simulation intervals that took account of the reliability data showed that the simulation interval grew wider as the sources of uncertainty taken into account grew more complete, but the associations expected a priori remained readily apparent. While many investigators use reliability data only as a metric for data quality, a more thorough approach can also quantitatively depict the uncertainty in the observed associations. PMID- 17406007 TI - Cardiorespiratory fitness as a predictor of nonfatal cardiovascular events in asymptomatic women and men. AB - Prospective data relating cardiorespiratory fitness (CRF) with nonfatal cardiovascular disease (CVD) events are limited to studies in men or studies of combined fatal and nonfatal CVD endpoints. The authors examined the association between CRF and nonfatal CVD events in 20,728 men and 5,909 women without CVD at baseline. All participants performed a maximal treadmill exercise test and completed a follow-up health survey in the Aerobics Center Longitudinal Study (Dallas, Texas) between 1971 and 2004. There were 1,512 events in men and 159 events in women during an average follow-up of 10 years. Across incremental CRF groups, age- and examination year-adjusted event rates per 10,000 person-years were 107.9, 75.2, and 50.3 in men (p(trend) < 0.001) and 41.9, 27.7, and 20.8 in women (p(trend) = 0.002). After further adjustment for smoking, alcohol intake, family history of CVD, and abnormal exercise electrocardiogram responses, hazard ratios were 1.00 (referent), 0.82 (95% confidence interval (CI): 0.72, 0.94), and 0.61 (95% CI: 0.53, 0.71) in men, p(trend) < 0.001, and were 1.00 (referent), 0.74 (95% CI: 0.49, 1.13), and 0.63 (95% CI: 0.40, 0.98) in women, p(trend) = 0.05. After adjustment for other CVD predictors, the association remained significant in men but not in women. PMID- 17406008 TI - Prenatal organophosphate metabolite and organochlorine levels and performance on the Brazelton Neonatal Behavioral Assessment Scale in a multiethnic pregnancy cohort. AB - Prenatal exposures to organophosphate pesticides and polychlorinated biphenyls have been associated with abnormal neonatal behavior and/or primitive reflexes. In 1998-2002, the Mount Sinai Children's Environmental Health Center (New York City) investigated the effects of indoor pesticide use and exposure to polychlorinated biphenyls on pregnancy outcome and child neurodevelopment in an inner-city multiethnic cohort. The Brazelton Neonatal Behavioral Assessment Scale was administered before hospital discharge (n = 311). Maternal urine samples were analyzed for six dialkylphosphate metabolites and malathion dicarboxylic acid. A random subset of maternal peripheral blood samples from the entire cohort (n = 194) was analyzed for polychlorinated biphenyls and 1,1'-dichloro-2,2'-bis(4 chlorophenyl)ethylene. Malathion dicarboxylic acid levels above the limit of detection were associated with a 2.24-fold increase in the number of abnormal reflexes (95% confidence interval: 1.55, 3.24). Likewise, higher levels of total diethylphosphates and total dialkylphosphates were associated with an increase in abnormal reflexes, as was total dimethylphosphates after paraoxonase expression was considered. No adverse associations were found with polychlorinated biphenyl or 1,1'-dichloro-2,2'-bis(4-chlorophenyl)ethylene levels and any behavior. The authors uncovered additional evidence that prenatal levels of organophosphate pesticide metabolites are associated with anomalies in primitive reflexes, which are a critical marker of neurologic integrity. PMID- 17406009 TI - Fat flies expanded the hippo pathway: a matter of size control. AB - By simultaneously inhibiting cell proliferation while promoting apoptosis, the Hippo signaling pathway provides a robust mechanism to restrict organ size during Drosophila development. Despite impressive progress in revealing the key intracellular components of this growth-regulatory pathway, the nature of the signal that regulates Hippo signaling in vivo has remained elusive. Several studies now implicate the atypical cadherin protein Fat as a cell surface receptor for the Hippo signaling pathway, thus potentially linking the Hippo kinase cascade with the extracellular milieu. PMID- 17406010 TI - Performance measurement of nursing care: state of the science and the current consensus. AB - Concerns about nurse staffing in hospitals, nursing's influence on patient safety and health care outcomes, and nurses' work environment (e.g., equipment failures, documentation burden) have led to increased interest in measuring and reporting nursing's performance. This article reviews recent efforts and issues involved in identifying a set of nursing-sensitive performance measures. Sustaining and strengthening current efforts requires developing measures that address all the domains of nursing, addressing technical issues needed to analyze the impact of nursing on patient safety and health care outcomes, developing data systems that provide the information needed to implement the model system, regularly improving the set of endorsed standards to reflect the most current science and empirical evidence, and persuading all health care stakeholders that measurement and reporting nursing-sensitive standards make a difference in the care and quality that are delivered. Each of these tasks requires substantial development work and construction and maintenance of the infrastructure to sustain the performance measurement efforts. PMID- 17406011 TI - Nursing sensitive databases: their existence, challenges, and importance. AB - The National Quality Forum uses the term "nurse-sensitive" care to describe health care quality that is influenced by nursing. Identifying this type of care requires useful data and development of measures. Establishing a case for nursing sensitive performance measurement will facilitate a supportive climate for research and measure development. Interdisciplinary investigations will result in more adequate measures and wider acceptance of the measures in the provider community. This article summarizes existing data sources and recommendations for improving measures, data collection, and research on nurse-sensitive care. PMID- 17406012 TI - Specifying and standardizing performance measures for use at a national level: implications for nursing-sensitive care performance measures. AB - This article explicates a comprehensive process for identifying, specifying, testing, and implementing nationally standardized performance measures. A growing body of research ties nurse staffing to patient outcomes, reinforcing the important role of nursing in the delivery of safe, efficacious health care. The Joint Commission has developed a multistep process to identify robust sets of evidence-based measures that includes a review of the evidence, expert advisory panel guidance, use of established criteria, public input and comment, and multistage field testing before implementation. This proven approach to the establishment of performance metrics is applicable to the identification and implementation of nurse-sensitive workforce measures. These metrics can then be used by providers to support performance improvement activities by health care stakeholders to monitor nurse staffing-related issues, and by researchers to better understand the relationship between staffing effectiveness and patient safety, and the quality of health care. PMID- 17406013 TI - Nursing measures: implementation considerations: lessons learned from the field. AB - With today's focus on performance measurement, pay for quality, and other data driven initiatives in the health care industry, measurement developers and researchers interested in testing these metrics need to keep in mind the needs of those end users that will be tasked with collecting, submitting, and in many cases analyzing the information that flows from these measures. This article will highlight a number of key issues learned from the field and suggest opportunities for research into the design and deployment of these measures. PMID- 17406014 TI - The nursing practice environment: measurement and evidence. AB - Improvements in nurses' practice environments are essential to retain nurses and keep patients safe. The pace of improvements can accelerate if evidence is translated clearly for researchers, managers, and policymakers. This article evaluates the utility of published multidimensional instruments to measure the nursing practice environment. The assessment criteria are theoretical relevance, ease of use, and dissemination. This article also synthesizes the research that has used these instruments. Seven instruments and 54 studies are evaluated. The Practice Environment Scale of the Nursing Work Index (PES-NWI) is proposed as the most useful instrument. Its content, length, and dissemination best satisfy the set of criteria. Researchers should use the PES-NWI to generate consistent and comparable evidence; expand the content to reflect all conceptual domains; develop a short form; test the instrument in different care settings; expand the evidence of the practice environment's influence on patient outcomes; and test interventions for practice environment improvements. PMID- 17406015 TI - The impact of nursing interventions: overview of effective interventions, outcomes, measures, and priorities for future research. AB - The purpose of this article is to present findings from a review of published systematic/integrative reviews and meta-analyses on nursing interventions and patient outcomes in acute care settings. A literature search was conducted for the period 1999-2005, producing 4,000 systematic/integrative reviews and 500 meta analyses covering seven topics selected by the authors: elder care, caregivers, developmental care of neonates and infants, symptom management, pressure ulcer prevention/treatment, incontinence, and staffing. The association between nursing care interventions/processes and patient outcomes in acute care settings was found to be limited in the articles reviewed. The strongest evidence was for the use of patient risk-assessment tools and interventions implemented by nurses to prevent patient harm. We observed significant variation in methods to measure the effect of independent variables (nursing interventions) on patient outcomes. Results indicate the need for more research measuring the effect of specific nursing interventions that may impact acute care patient outcomes. PMID- 17406016 TI - Advancing the science in the measurement of health care quality influenced by nurses. AB - A robust set of quality measures is essential to provide consumers with a vehicleto evaluate nurses' contributions to the care of hospitalized patients, providers, and systems with a set of nursing processes and outcomes to guide quality improvement, and insurers with indicators to reward hospitals for high quality nursing services. The processes employed by the Nursing Care Performance Measures Steering Committee convened by the National Quality Forum (NQF) in 2004 resulted in the endorsement of 15 indicators of health care quality influenced by nurses and contributed to the identification of significant gaps in measurement and priority areas for future research. This critical review of the state of the science related to health care processes and outcomes that reflect nurses' contributions to the quality of care for hospitalized patients is intended to push the boundaries in the measurement of nursing performance. Specific recommendations for future research and measure development are presented. PMID- 17406017 TI - Information and communications technology in chronic disease care: what are the implications for payment? AB - New information and communication technology (ICT) has the potential to improve care for chronic conditions, deliver better health outcomes, and reduce direct and indirect costs. However, realizing these gains necessitates new forms of care delivery, through adoption of ICT, the specific choice of ICT, and changes to existing forms of health care delivery. Realizing these new forms of delivery requires appropriate payment policies. The authors review the literatures on health care payment and ICT in chronic care and then apply theoretical economic analysis to determine how ICT alters health care payment policy recommendations. Using diabetes as an example, the authors identify and illustrate the disease and technology features that determine the optimal form of payment. Overall, ICT shifts the optimal blend of fee-for-service and capitation toward greater capitation. Carve-outs for ICT-supported preventive care enable more high-powered payment while addressing selection concerns. PMID- 17406018 TI - Hospital financial condition and operational decisions related to the quality of hospital care. AB - Financial pressure mounted for hospitals nationwide during the late 1990s. Our study examines how this affected the quality of their operations in terms of organizational infrastructure and processes that support the delivery of care. Our sample consisted of community hospitals operating between 1995 and 2000. Financial pressure was measured based on changes in net patient revenues per adjusted patient day and the ratio of cash flow to total revenues. The authors examined effects on hospital investments in plant and equipment and on hospital standards compliance with selected Joint Commission on Accreditation of Healthcare Organization performance areas. The results suggest that increasing financial pressures did lead to cutbacks in these areas. These findings suggest the importance of looking broadly across hospital operations to identify factors that may contribute to poor patient outcomes. Given the findings of earlier studies, these results suggest that poor outcomes may in part result from deterioration in supporting infrastructure and organizational processes. PMID- 17406019 TI - Less is more in presenting quality information to consumers. AB - Much effort has been put into improving measures of health care quality. Although early research suggested that consumers made little use of quality reports, most reports were based on nonstandardized measures and were not user friendly. Information presentation approaches, however, will have a significant influence on what information is attended and used. The present research examines whether information presentation methods differentially influence consumers who differ in numeric skills. Results of three studies support the idea that "less is more" when presenting consumers with comparative performance information to make hospital choices. Results were particularly strong for those lower in numeracy, who had higher comprehension and made better choices when the information presentation format was designed to ease the cognitive burden and highlight the meaning of important information. These findings have important implications for the sponsors of comparative quality reports designed to inform consumer decision making in health care. PMID- 17406020 TI - Job satisfaction of nursing home administrators and turnover. AB - We know little about factors associated with job satisfaction and dissatisfaction for nursing home workers. In this investigation, the authors use data from a large sample of nursing home administrators (NHAs) to examine: (1) their levels of job satisfaction, (2) whether job satisfaction is associated with intent to leave, (3) whether job satisfaction is associated with turnover after 1 year, and (4) whether job satisfaction after 1 year varies for NHAs who left based on where they subsequently worked. Overall, NHAs were more satisfied with the job satisfaction subscales of: rewards, work skills, and workload but were less satisfied with work demands and coworkers. NHAs appeared particularly sensitive to work skills, with this area of job satisfaction being associated with intent to turnover and actual turnover. In general, the authors found a stronger association with job satisfaction and actual turnover than with intent to turnover. PMID- 17406021 TI - Financial protection afforded by employer-sponsored health insurance: current plan designs and high-deductible health plans. AB - This article provides the first national estimates of actuarial values and out-of pocket spending from the era of nonrestrictive managed care that began in the late 1990s. Employer plans paid about 84 percent of total medical expense for those with employer-sponsored coverage in 2004, about 1 percent less than in 2000, and high users faced potential out of pocket spending in the thousands of dollars when they received a portion of their care out of network. Since 2004, more employers have offered plans with higher deductibles coupled with employer funded personal accounts. These arrangements can result in low out of pocket costs for many employees, but high users will face substantially higher costs. Many employers adopting high-deductible plans are not contributing to personal accounts. Those who are concerned about higher out-of-pockets might consider income-related cost sharing, educational efforts to communicate the savings that can result from using in-network providers, and continued availability of managed care options that limit out-of-pocket spending. PMID- 17406022 TI - Testing the neutral fixation of hetero-oligomerism in the archaeal chaperonin CCT. AB - The evolutionary transition from homo-oligomerism to hetero-oligomerism in multimeric proteins and its contribution to function innovation and organism complexity remain to be investigated. Here, we undertake the challenge of contributing to this theoretical ground by investigating the hetero-oligomerism in the molecular chaperonin cytosolic chaperonin containing tailless complex polypeptide 1 (CCT) from archaea. CCT is amenable to this study because, in contrast to eukaryotic CCTs where sub-functionalization after gene duplication has been taken to completion, archaeal CCTs present no evidence for subunit functional specialization. Our analyses yield additional information to previous reports on archaeal CCT paralogy by identifying new duplication events. Analyses of selective constraints show that amino acid sites from 1 subunit have fixed slightly deleterious mutations at inter-subunit interfaces after gene duplication. These mutations have been followed by compensatory mutations in nearby regions of the same subunit and in the interface contact regions of its paralogous subunit. The strong selective constraints in these regions after speciation support the evolutionary entrapment of CCTs as hetero-oligomers. In addition, our results unveil different evolutionary dynamics depending on the degree of CCT hetero-oligomerism. Archaeal CCT protein complexes comprising 3 distinct classes of subunits present 2 evolutionary processes. First, slightly deleterious and compensatory mutations were fixed neutrally at inter-subunit regions. Second, sub-functionalization may have occurred at substrate-binding and adenosine triphosphate-binding regions after the 2nd gene duplication event took place. CCTs with 2 distinct types of subunits did not present evidence of sub functionalization. Our results provide the 1st in silico evidence for the neutral fixation of hetero-oligomerism in archaeal CCTs and provide information on the evolution of hetero-oligomerism toward sub-functionalization in archaeal CCTs. PMID- 17406023 TI - Two infA gene mutations independently originated from a mutator genotype in barley. AB - Cytoplasmic line 2 (CL2) is a chlorophyll mutant that was selected from a plastid mutator genotype in barley. The dynamics of greening and plastid development of CL2 first-leaf blade contrasts with that of monocots. Previous characterizations of CL2 suggested that this mutant has a delay of plastid gene translation during embryogenesis. We hypothesize that CL2 is a mutant in the infA gene, which encodes translation initiation factor 1 (IF1). Wild-type barley infA gene differs in some nucleotides from that in wheat, but the corresponding IF1 proteins are identical. However, infA from CL2 carries a point mutation, which leads to an amino acid change in IF1 residue 52. One CL2-like seedling selected from a new mutator pool also carries a point mutation in infA gene, this time leading to a change of the universally conserved amino acid residue 32. Both point mutations were T --> C substitutions. We sequenced the complementary DNA of the infA transcripts from the wild type and CL2 and found that the mutation was conserved at the mRNA level. Results strongly suggest that CL2 and CL2-like are infA gene mutants, this being the first time that a mutant phenotype is attributed to infA gene in a higher plant. PMID- 17406024 TI - The role of induced mutation in conversion of photoperiod dependence in cotton. AB - Wild cotton germplasm resources are largely underutilized because of photoperiod dependent flowering of "exotic" cottons. The objectives of this work were to explore the genome-wide effect of induced mutation in photoperiod-converted induced cotton mutants, estimating the genetic change between mutant and wild type cottons using simple sequence repeats (SSRs) as well as understand the pattern of SSR mutation in induced mutagenesis. Three groups of photoperiod converted radiomutants ((32)P) including their wild-type parental lines, A- and D genome diploids, and typically grown cotton cultivars were screened with 250 cotton SSR primer pairs. Forty SSRs revealed the same SSR mutation profile in, at least, 2 independent mutant lines that were different from the original wild types. Induced mutagenesis both increased and decreased the allele sizes of SSRs in mutants with the higher mutation rate in SSRs containing dinucleotide motifs. Genetic distance obtained based on 141 informative SSR alleles ranged from 0.09 to 0.60 in all studied cotton genotypes. Genetic distance within all photoperiod converted induced mutants was in a 0.09-0.25 range. The genetic distance among photoperiod-converted mutants and their originals ranged from 0.28 to 0.50, revealing significant modification of mutants from their original wild types. Typical Gossypium hirsutum cultivar, Namangan-77, revealed mutational pattern similar to induced radiomutants in 40 mutated SSR loci, implying possible pressure to these SSR loci not only in radiomutagenesis but also during common breeding process. Outcomes of the research should be useful in understanding the photoperiod-related mutations, and markers might help in mapping photoperiodic flowering genes in cotton. PMID- 17406025 TI - An extended microsatellite set for linkage mapping in the domestic dog. AB - The extremes of phenotype displayed by the domestic dog, as well as the largest number of naturally occurring inherited diseases in any mammalian species except man (>450), have generated a large interest in genomic linkage mapping in the species. Marker sets for linkage mapping should ideally show both high levels of polymorphism among the target group of animals and an even spacing of markers across the whole genome. Currently a microsatellite marker set known as Minimal Screening Set 2 (MSS2) is widely used. Here, we have extended this marker set by filling in gaps as noted from the marker positions in the CanFam genome assembly (1.0) and the 5000cR radiation hybrid (RH) map. An additional 183 markers have been positioned to increase the coverage of the MSS2 set wherever it contains a gap >9 mb or 1000(5000) RH units. We have called the marker set derived from the MSS2 set and these 183 markers, MSS3. The average physical spacing of markers in the complete 507 marker MSS3 set is 5 mb, whereas average heterozygosity of the 183 new markers on a panel of 10 dogs of differing breeds is 0.74. This marker group will allow genome-wide scans in the dog to be conducted at close to 5 cM resolution. PMID- 17406026 TI - MUC4 mucin potentiates pancreatic tumor cell proliferation, survival, and invasive properties and interferes with its interaction to extracellular matrix proteins. AB - MUC4, a transmembrane mucin, is aberrantly expressed in pancreatic adenocarcinomas while remaining undetectable in the normal pancreas. Recent studies have shown that the expression of MUC4 is associated with the progression of pancreatic cancer and is inversely correlated with the prognosis of pancreatic cancer patients. In the present study, we have examined the phenotypic and molecular consequences of MUC4 silencing with an aim of establishing the mechanistic basis for its observed role in the pathogenesis of pancreatic cancer. The silencing of MUC4 expression was achieved by stable expression of a MUC4 specific short hairpin RNA in CD18/HPAF, a highly metastatic pancreatic adenocarcinoma cell line. A significant decrease in MUC4 expression was detected in MUC4-knockdown (CD18/HPAF-siMUC4) cells compared with the parental and scrambled short interfering RNA-transfected (CD18/HPAF-Scr) control cells by immunoblot analysis and immunofluorescence confocal microscopy. Consistent with our previous observation, inhibition of MUC4 expression restrained the pancreatic tumor cell growth and metastasis as shown in an orthotopic mouse model. Our in vitro studies revealed that MUC4-associated increase in tumor cell growth resulted from both the enhanced proliferation and reduced cell death. Furthermore, MUC4 expression was also associated with significantly increased invasiveness (P < or = 0.05) and changes in actin organization. The presence of MUC4 on the cell surface was shown to interfere with the tumor cell-extracellular matrix interactions, in part, by inhibiting the integrin-mediated cell adhesion. An altered expression of growth- and metastasis-associated genes (LI-cadherin, CEACAM6, RAC1, AnnexinA1, thrombomodulin, epiregulin, S100A4, TP53, TP53BP, caspase-2, caspase-3, caspase-7, plakoglobin, and neuregulin-2) was also observed as a consequence of the silencing of MUC4. In conclusion, our study provides experimental evidence that supports the functional significance of MUC4 in pancreatic cancer progression and indicates a novel role for MUC4 in cancer cell signaling. PMID- 17406027 TI - Multiple molecular markers as predictors of colorectal cancer in patients with normal perioperative serum carcinoembryonic antigen levels. AB - PURPOSE: In this study, a high-sensitivity colorimetric membrane array method was used to detect circulating tumor cells (CTC) in the peripheral blood of colorectal cancer (CRC) patients with normal perioperative serum carcinoembryonic antigen (CEA) levels. This membrane array method was evaluated as a potential diagnostic and postoperative surveillance tool. STUDY DESIGN: Membrane arrays consisting of a panel of mRNA markers that include human telomerase reverse transcriptase, cytokeratin-19, cytokeratin-20, and CEA mRNA were used to detect CTCs in the peripheral blood of 157 postoperative CRC patients with normal perioperative serum CEA levels and in 80 healthy individuals. Digoxigenin-labeled cDNA were amplified by reverse transcription-PCR from the peripheral blood samples, which were then hybridized to the membrane array. The sensitivity, specificity, and accuracy of membrane arrays for the detection of CTCs were then calculated. RESULTS: Using the four markers in combination, expression of any three markers or all the four markers in this panel was significantly correlated with the clinicopathologic characteristics, including depth of tumor invasion, lymph node metastasis, tumor-node-metastasis stage, and postoperative relapse (all P < 0.05). The interval between the detection of all four positive molecular markers and subsequent elevated CEA ranged from 3 to 8 months (median 6 months). The expression of all four mRNA markers was an independent predictor for postoperative relapse. CRC patients with all four mRNA markers expression showed a significantly poorer survival rate than those with less than four positive markers. CONCLUSIONS: The constructed membrane array method was helpful in the early prediction of postoperative relapse in CRC patients with normal perioperative serum CEA levels. PMID- 17406028 TI - Down-regulation of CD9 expression during prostate carcinoma progression is associated with CD9 mRNA modifications. AB - PURPOSE: Cluster-of-differentiation antigen 9 (CD9) protein, a member of the tetraspanin family, has been implicated in carcinogenesis of various human tumors. Although decreased expression of the CD82 tetraspanin protein, a close CD9 relative, is associated with prostate cancer progression, CD9 expression has not been analyzed in this malignancy. EXPERIMENTAL DESIGN: CD9 expression in human prostatic adenocarcinoma was analyzed by immunohistochemistry on 167 primary tumors and 88 lymph node or bone metastases. CD9 cDNA was sequenced from two human prostate cancer cell lines, prostatic adenocarcinoma, high-grade prostatic intraepithelial neoplasia (PIN), and normal prostatic tissues. RESULTS: Although CD9 was detected in the epithelium of normal prostatic tissues, reduced or loss of CD9 expression within neoplastic cells was observed in 24% of 107 clinically localized primary adenocarcinomas, 85% of 60 clinically advanced primary adenocarcinomas, 85% of 65 lymph node metastases, and 65% of 23 bone metastases. Difference in CD9 expression between clinically localized and advanced diseases was highly significant (P < 1 x 10(-7)). Whereas there was no alteration of CD9 cDNA in normal tissues, all PC-3-derived cell lines, one PIN, and four prostatic adenocarcinomas harbored deletions in their CD9 cDNAs. Recurring CD9 point mutations were also found in PC-3M-LN4 cells, one PIN, and seven prostatic adenocarcinomas. CONCLUSIONS: CD9 expression is significantly reduced and even lost during prostate cancer progression. Moreover, deletions and mutations of the CD9 mRNA may be associated with loss of protein expression observed in tumor cells. Our data suggest that CD9 inactivation may play an important role in prostate cancer progression. PMID- 17406029 TI - Predicting gefitinib responsiveness in lung cancer by fluorescence in situ hybridization/chromogenic in situ hybridization analysis of EGFR and HER2 in biopsy and cytology specimens. AB - In non-small cell lung cancer (NSCLC), epidermal growth factor receptor (EGFR) mutational analysis is an excellent predictor of responsiveness to treatment with tyrosine kinase inhibitors, such as gefitinib. In up to 80% of NSCLCs, cytologic samples or endoscopic biopsies are the only specimens available for molecular analysis, but PCR amplification of DNA from small fixed and paraffin-embedded samples may create artifactual mutations. Fluorescence in situ hybridization (FISH) of EGFR and HER2 has been proposed as an alternative method of analysis. This project aimed to determine the optimal scoring method for FISH or chromogenic in situ hybridization (CISH) assays when analyzing small NSCLC samples to predict response. FISH or CISH analysis of EGFR and HER2 genes was done on 42 small samples derived from NSCLC patients treated with gefitinib. EGFR mutational analysis was done after quantity and quality controls of DNA. In seven of seven cases, a balanced increase in EGFR gene and chromosome 7 number was found to correlate with the presence of specific EGFR mutations. In addition, seven of seven cases with balanced EGFR/HER2 polysomy and two of three cases with balanced EGFR/HER2 trisomy responded to gefitinib (75% of responders). Instead, the EGFR mutations predicted only 7 of 12 (58%) of gefitinib-responsive patients. When only endoscopic biopsies or cytologic specimens are available, we propose using FISH/CISH for EGFR and HER2 as the test of choice for selecting patients for treatment with gefitinib and to consider as negative predictive factor the absence of EGFR/HER2 gene gain. PMID- 17406030 TI - MAP-ing glioma invasion: mitogen-activated protein kinase kinase 3 and p38 drive glioma invasion and progression and predict patient survival. AB - Although astrocytic brain tumors do not metastasize systemically, during tumorigenesis glioma cells adopt an invasive phenotype that is poorly targeted by conventional therapies; hence, glioma patients die of recurrence from the locally invasive tumor population. Our work is aimed at identifying and validating novel therapeutic targets and biomarkers in invasive human gliomas. Transcriptomes of invasive glioma cells relative to stationary cognates were produced from a three dimensional spheroid in vitro invasion assay by laser capture microdissection and whole human genome expression microarrays. Qualitative differential expression of candidate invasion genes was confirmed by quantitative reverse transcription-PCR, clinically by immunohistochemistry on tissue microarray, by immunoblotting on surgical specimens, and on two independent gene expression data sets of glial tumors. Cell-based assays and ex vivo brain slice invasion studies were used for functional validation. We identify mitogen-activated protein kinase (MAPK) kinase 3 (MKK3) as a key activator of p38 MAPK in glioma; MKK3 activation is strongly correlated with p38 activation in vitro and in vivo. We further report that these members of the MAPK family are strong promoters of tumor invasion, progression, and poor patient survival. Inhibition of either candidate leads to significantly reduced glioma invasiveness in vitro. Consistent with the concept of synthetic lethality, we show that inhibition of invasion by interference with these genes greatly sensitizes arrested glioma cells to cytotoxic therapies. Our findings therefore argue that interference with MKK3 signaling through a novel treatment combination of p38 inhibitor plus temozolomide heightens the vulnerability of glioma to chemotherapy. PMID- 17406031 TI - Tumor-stroma interactions in pancreatic ductal adenocarcinoma. AB - The host stromal response to an invasive epithelial carcinoma is frequently called a desmoplastic reaction (DR) and is a universal feature of pancreatic ductal adenocarcinoma (PDA). This DR is characterized by a complex interplay between the normal host epithelial cells, invading tumor cells, stromal fibroblasts, inflammatory cells, proliferating endothelial cells, an altered extracellular matrix, and growth factors activating oncogenic signaling pathways by autocrine and paracrine mechanisms. Hence, the tumor microenvironment is a dynamic process promoting tumor growth and invasion through mechanisms likely to include anoikis resistance, genomic instability, and drug resistance. Cell coculture models, murine models (xenograft and genetic), and gene expression profiling studies on human PDA biopsies have identified several key molecules, such as collagen type I, fibronectin, laminin, matrix metalloproteinases (MMP) and their inhibitors (tissue inhibitors of MMP), growth factors (transforming growth factor beta, platelet-derived growth factor, connective tissue growth factor, and hepatocyte growth factor), chemokines, and integrins as constituents of the DR. Despite these findings, it is unclear which molecular-cellular events initiate and drive desmoplasia in PDA. Accumulating evidence indicates that pancreatic stellate cells when activated switch to a myofibroblast phenotype that produces components of the extracellular matrix, MMPs, and tissue inhibitors of MMPs by activating the mitogen-activated protein kinase (extracellular signal regulated kinase 1/2) pathway. Based on current evidence, several therapeutic strategies are been evaluated on identified potential therapeutic targets. This review summarizes our current understanding of the mechanisms that potentially drive the DR in PDA and future possibilities for therapeutic targeting of this critical process. PMID- 17406032 TI - H2AX phosphorylation marks gemcitabine-induced stalled replication forks and their collapse upon S-phase checkpoint abrogation. AB - Gemcitabine is a nucleoside analogue that is incorporated into replicating DNA, resulting in partial chain termination and stalling of replication forks. The histone variant H2AX is phosphorylated on Ser(139) (gamma-H2AX) and forms nuclear foci at sites of DNA damage. Here, we characterize the concentration- and time dependent phosphorylation of H2AX in response to gemcitabine-induced stalled replication forks. The number of gamma-H2AX foci increased with time up to 2 to 6 h after exposure to gemcitabine, whereas longer exposures did not cause greater phosphorylation or increase cell death. The percentage of gamma-H2AX-positive cells increased with concentrations of gemcitabine up to 0.1 micromol/L, and gamma-H2AX was most evident in the S-phase fraction. Phosphorylation of ataxia telangiectasia mutated (ATM) on Ser(1981) was also associated with S-phase cells and colocalized in the nucleus with phosphorylated H2AX foci after gemcitabine exposure. Chemical inhibition of ATM, ATM- and Rad3-related, and DNA-dependent protein kinase blocked H2AX phosphorylation. H2AX and ATM phosphorylation were associated with inhibition of DNA synthesis, S-phase accumulation, and activation of the S-phase checkpoint pathway (Chk1/Cdc25A/cyclin-dependent kinase 2). Exposure of previously gemcitabine-treated cultures to the Chk1 inhibitor 7 hydroxystaurosporine (UCN-01) caused a 10-fold increase in H2AX phosphorylation, which was displayed as an even pan-nuclear staining. This increased phosphorylation was not due to apoptosis-induced DNA fragmentation and was associated with the S-phase fraction and decreased reproductive viability. Thus, H2AX becomes phosphorylated and forms nuclear foci in response to gemcitabine induced stalled replication forks, and this is greatly increased upon checkpoint abrogation. PMID- 17406033 TI - Activation of a novel ataxia-telangiectasia mutated and Rad3 related/checkpoint kinase 1-dependent prometaphase checkpoint in cancer cells by diallyl trisulfide, a promising cancer chemopreventive constituent of processed garlic. AB - Diallyl trisulfide (DATS), a cancer chemopreventive constituent of garlic, inhibits growth of cancer cells by interfering with cell cycle progression, but the mechanism is not fully understood. Here, we show the existence of a novel ataxia-telangiectasia mutated and Rad3 related (ATR)/checkpoint kinase 1 (Chk1) dependent checkpoint partially responsible for DATS-mediated prometaphase arrest in cancer cells, which is different from the recently described gamma irradiation induced mitotic exit checkpoint. The PC-3 human prostate cancer cells synchronized in prometaphase by nocodazole treatment and released to DATS containing medium remained arrested in prometaphase, whereas the cells released to normal medium exited mitosis and resumed cell cycle. The mitotic arrest was maintained even after 4 h of culture of DATS-treated cells (4-h treatment) in drug-free medium. The DATS-arrested mitotic cells exhibited accumulation of anaphase-promoting complex/cyclosome (APC/C) substrates cyclin A and cyclin B1 and hyperphosphorylation of securin, which was accompanied by increased phosphorylation of the APC/C regulatory subunits Cdc20 and Cdh1. The DATS mediated accumulation of cyclin B1 and hyperphosphorylation of securin, Cdc20, and Cdh1 were partially but markedly attenuated by knockdown of Chk1 or ATR protein. The U2OS osteosarcoma cells expressing doxycycline-inducible kinase dead ATR were significantly more resistant not only to DATS-mediated prometaphase arrest but also to the accumulation of cyclin B1 and hyperphosphorylation of securin, Cdc20, and Cdh1 compared with cells expressing wild-type ATR. However, securin protein knockdown failed to rescue cells from DATS-induced prometaphase arrest. In conclusion, the present study describes a novel signaling pathway involving ATR/Chk1 in the regulation of DATS-induced prometaphase arrest. PMID- 17406034 TI - The stroke epidemic. PMID- 17406035 TI - Outcomes after first-ever stroke. AB - OBJECTIVES: To determine the outcomes after first-ever stroke, including mortality, dependence, and recurrence. DESIGN: Retrospective study on a prospectively collected cohort. SETTING: Regional hospital, Hong Kong. PATIENTS: A cohort of 755 patients presented to our hospital from 1996 to 1998 with their first-ever stroke. MAIN OUTCOME MEASURES: Mortality and stroke recurrence rate at 30 days, 1 year, and 5 years from the onset of the stroke. Dependence in activity of daily living at 5 years from the onset of stroke. RESULTS: The mortality rate was 15.1% at 30 days, 22.5% at 1 year, and 39.7% at 5 years from the onset of the first-ever stroke. The rate of stroke recurrence was 0.9% at 30 days, 7.0% at 1 year, and 21.2% at 5 years from the onset of first-ever stroke. Among patients presenting with ischaemic strokes, 109 (20.6%) had a recurrence, of which 92 (84%) were ischaemic strokes and 17 (16%) were haemorrhagic. Among patients presenting with intracerebral haemorrhage, 25 (23.1%) had a recurrence, of which 12 (48%) were haemorrhagic strokes and 13 (52%) patients were ischaemic. After 5 years, 11% of the patients were dependent in terms of activity of daily living. CONCLUSIONS: The long-term prognosis after first-ever stroke is poor--5 years after their stroke, 39.7% of patients had died and 10.7% were dependent in terms of activity of daily living; 136 (21%) who survived at least 30 days after the initial stroke, had a recurrence within 5 years. PMID- 17406036 TI - Determinants of preference for elective caesarean section in Hong Kong Chinese pregnant women. AB - OBJECTIVE: To find the clinical and socio-demographic determinants for Hong Kong Chinese women who preferred elective caesarean section. DESIGN: Cross-sectional interview survey. SETTING: University teaching hospital, Hong Kong. PARTICIPANTS: A cohort of consecutive Hong Kong Chinese pregnant women (n=660) attending a government-funded obstetric unit catering deliveries in the New Territories in Hong Kong in 2002. MAIN OUTCOME MEASURES: The clinical and socio-demographic determinants of preference for elective caesarean section, in women who could have a trial of vaginal delivery. RESULTS: The overall prevalence for maternal preference for elective caesarean section was 16.7% (95% confidence interval, 13.8-19.6). The factors associated with preferring elective caesarean section were: previous elective caesarean section (odds ratio=7.6; 95% confidence interval, 2.0-28.7) and previous emergency caesarean section (3.8; 1.8-8.2). Among nulliparous women, the prevalence of preference for elective caesarean section was 16.8% (95% confidence interval, 13.0-20.6). Conception by in-vitro fertilisation was found to be significantly associated with preferring elective caesarean section in nulliparous women (odds ratio=5.2; 95% confidence interval, 1.0-26.4). CONCLUSION: Previous caesarean section and conception by in-vitro fertilisation were determinants for women preferring elective caesarean section. PMID- 17406037 TI - Opportunistic breast cancer screening in Hong Kong; a revisit of the Kwong Wah Hospital experience. AB - OBJECTIVE: To analyse the performance of opportunistic breast screening in local Well Women Clinics during the 5-year period from 1998 to 2002, with reference to international as well as our own earlier experience (1991 to 1993 to 1995). DESIGN: Prospective study. SETTING: Well Women Clinics in regional Hong Kong hospitals. PARTICIPANTS: Women attending the Well Women Clinics of the Tung Wah Group of Hospitals for breast cancer screening. MAIN OUTCOME MEASURES: All screening-detected breast cancers. RESULTS: After 46 637 screening mammograms and excluding palpable masses detected by the patients themselves, 232 women were detected with cancers, yielding a crude detection rate of 5.0 per 1000. Age range for cancer detection was 35 to 72 (median, 49) years. Clinic staff detected palpable lumps in 83 patients, constituting 36% of the detected cancers, of which 15 (6.5% of all detected cancers) were not identified by mammography. The cancer detection rate was 5.9 per 1000 in the age-group 40-49 years and 3.7 per 1000 in those 50 years or older. The cancer detection rate was 58.5 per 1000 in the high risk group (aged <40 years with positive family history). The minimal cancer detection rate was 2.2 per 1000, representing 45% of all cancers whose pathology was available to us. Ductal carcinoma in situ comprised 28% of all such cancers. Our recall rates were 4.6% for additional views only, 3.3% for ultrasound only, and 2.3% for both. Positive predictive values for abnormal mammograms and biopsies were 4.9% and 26.0%, respectively. CONCLUSION: Mammographic screening has been reported to reduce mortality up to 35% in western countries. However, data concerning Asian Chinese populations are meagre. Our Well Women Clinics pioneered large-scale self-referred breast screening in Hong Kong. Despite the lower incidence of breast cancers than in Occidental populations, our screening programme performed comparably to those in the West, and has improved over time. Our screening service for self-referred women detected breast cancers even at an early stage, which facilitated a better prognosis and more treatment options, whilst appearing to be highly acceptable to our community. Besides, it could provide training and expertise in breast radiology and mammography, which are essential prerequisites for establishing population screening. PMID- 17406038 TI - Assessing disease burden of respiratory disorders in Hong Kong children with hospital discharge data and linked laboratory data. AB - OBJECTIVES: To describe the pattern of respiratory disorders in the Hong Kong paediatric population admitted to government hospitals, and to assess the reliability of the diagnoses by linkage with laboratory data. METHODS: Discharge diagnoses for all admissions are recorded in a central computerised database, the Clinical Management System. These data were analysed for the inclusive period July 1997 to June 1999. Virology laboratory results from a single hospital were linked to the Clinical Management System diagnostic codes to examine discrepancies in coding specific viral aetiologies. RESULTS: A primary diagnosis of a respiratory disorder was noted in 37.5% (upper respiratory 30.1%, tonsillitis/pharyngitis 10.5%, croup/laryngitis 2.3%, acute otitis media 2.7%, bronchitis/chest infection 2.6%, bronchiolitis 10.2%, pneumonia 20.9%, influenza 4%, asthma and allergic rhinitis 16.5%), and a primary or secondary diagnosis in 42.5% of children younger than 15 years. The incidence rates of respiratory illness coded as bronchiolitis and influenza were respectively estimated to be 887-979 and 222-381 per 100,000 children under 5 years and 3551-3949 and 415-528 per 100,000 children under the age of 1 year. The percentage of respiratory associated admissions varied significantly by hospital and detailed analysis of data at one hospital highlighted important discrepancies between discharge diagnosis and laboratory results. CONCLUSIONS: These passive surveillance data provide general estimates of the disease burden for respiratory disorders in Hong Kong children. Active surveillance studies are required to provide more accurate estimates of the disease burden. Consideration should be given to enhance the Clinical Management System by routinely linking all laboratory data with discharge diagnosis information, by establishing sentinel surveillance hospitals and by assessing new strategies to standardise coding. PMID- 17406039 TI - Aortic dissection in an accident and emergency department in Hong Kong. AB - OBJECTIVES: To compare the clinical characteristics of aortic dissection in Hong Kong with the International Registry of Aortic Dissection, and to evaluate the sensitivity of the Accident and Emergency Department in diagnosing aortic dissection and its role in aortic dissection management. DESIGN: Retrospective observational study. SETTING: Regional public hospital, Hong Kong. PATIENTS: Newly diagnosed aortic dissection patients attending the Accident and Emergency Department from 2002 to 2005 inclusive. MAIN OUTCOME MEASURES: Correct diagnosis in Accident and Emergency Department, tertiary unit transfer, and in-patient mortality. RESULTS: Twenty-eight patients were found to have aortic dissection, with an estimated annual incidence of 2.1 per 100,000 inhabitants. The sensitivity of Accident and Emergency Department in diagnosing aortic dissection was 54%; 11% of the patients were diagnosed at postmortem examination. Compared to the International Registry of Aortic Dissection, the patients in this Tseung Kwan O Hospital study had less abrupt and less severe pain, less chest pain, and a lower proportion were operated on. Higher mortality was associated with age 70 years or older (odds ratio=6.4), female gender (21.0), known hypertension (3.8), systolic blood pressure below 100 mm Hg (6.0), aortic dissection not diagnosed in the Accident and Emergency Department (3.2), and the patient not reaching tertiary unit (33.8). The hourly cumulative mortality rate was 1.32%. The group of aortic dissections diagnosed in the Accident and Emergency Department had 55.1% more transfer to tertiary unit (95% confidence interval, 14.4-79.1%; P=0.006), 84.5 hours less transfer time (95.3-263.6 hours; P=0.232), and 27.2% lower mortality (12.6-58.6%; P=0.246). The yield rate of contrast computed tomography of thorax was 43%. CONCLUSIONS: Diagnosing aortic dissection in the Accident and Emergency Department enabled optimal disposition and lower in patient mortality. PMID- 17406040 TI - The skin prick test results of 977 patients suffering from chronic rhinitis in Hong Kong. AB - OBJECTIVES: To evaluate the prevalence of aetiological allergens identifiable by skin prick test alone in patients suffering from chronic rhinitis in Hong Kong, and also compare the clinical history and symptoms of skin prick test-positive versus skin prick test-negative patients. DESIGN: Prospective study. SETTING: Otorhinolaryngology clinic in Queen Mary Hospital of Hong Kong. PATIENTS: A total of 977 patients suffering from chronic rhinitis were recruited into the study. Skin prick test was performed with a panel of allergens including house dust mites, cockroach, cat, dog, moulds, and pollens. MAIN OUTCOME MEASURES: Skin prick test results and their correlation with symptoms. RESULTS: Of the 977 patients, 651 (67%) had positive skin prick test reactions. The commonest allergen was house dust mite which was positive in 63% of the 977 patients and 95% of those 651 skin prick test-positive patients. The other allergens were in order of cockroach (23%), cat (14%), dog (5%), pollen (4%), and mould (3%). Compared with skin prick test-negative patients, skin prick test-positive patients were more likely to have earlier age of onset of the chronic rhinitis, association with asthma, more severe symptom in the morning, more severe symptoms of itchy nose, sneezing, nasal discharge, itchy eye, and watery eye. CONCLUSIONS: Identifiable aeroallergens could be detected in 67% chronic rhinitis patients by skin prick test alone. House dust mites were the most prevalent causative allergen. There were significant differences of patterns of clinical history and symptoms severity between skin prick test-positive and skin prick test-negative patients. PMID- 17406041 TI - Breast pathology in complications associated with polyacrylamide hydrogel (PAAG) mammoplasty. AB - OBJECTIVE: To study the tissue pathology of breast lesions associated with hydrophilic polyacrylamide gel injection augmentation mammoplasty. DESIGN: Retrospective study. SETTING: Private anatomical pathology practice, Hong Kong. PATIENTS: Eight patients who underwent lumpectomy of the breast due to complications of hydrophilic polyacrylamide gel injection for augmentation mammoplasty. MAIN OUTCOME MEASURES: Identification of hydrophilic polyacrylamide gel in breast tissue and associated pathological changes. RESULTS: We reviewed the pathological changes in breast tissue associated with hydrophilic polyacrylamide gel injection in eight cases retrieved from our archive. Microscopically, the hydrophilic polyacrylamide gel appeared as pools of pale violet gelatinous material of variable size, between the interstices of connective tissue and fat cells. The larger pools were often surrounded by cellular reactions consisting of histiocytic cells and foreign body-type multinucleated giant cells. Inflammatory reaction featuring infiltration by lymphocytes and plasma cells in the adjacent breast tissue was observed in samples from four patients, and a sample from another patient showed acute inflammation with abscess formation. There was no evidence of abnormal cellular proliferation, atypia or malignant change in the stromal connective tissue or ductal-acinic epithelial components of the breast tissue. CONCLUSIONS: Hydrophilic polyacrylamide gel injection for augmentation mammoplasty can give rise to a breast lump and inflammation. Pathologically, this complication is associated with fibrosis, foreign body reaction, and inflammation. PMID- 17406042 TI - A report of four cases of caesarean scar pregnancy in a period of 12 months. AB - We report on four cases of caesarean scar pregnancy with different modes of treatment--expectant, surgical, systemic medical, and local medical. We attempt to explore the indications, and pros and cons of the various management modalities for caesarean scar pregnancy. PMID- 17406043 TI - Spontaneous basilar artery dissection. AB - Dissection of the basilar artery is rare. We report a 51-year-old man who developed acute pontine infarction associated with dissection of the distal basilar artery. There was no trauma or unaccustomed movement of the head and neck prior to the stroke. The dissection was diagnosed non-invasively by magnetic resonance imaging and magnetic resonance angiography. Cervicocerebral artery dissection is a common cause of stroke in young patients particularly when conventional cardiovascular risk factors are absent. Magnetic resonance angiography combined with magnetic resonance imaging is a useful diagnostic tool prior to invasive angiography. PMID- 17406044 TI - Supradiaphragmatic and transdiaphragmatic intrathoracic migration of a ventriculoperitoneal shunt catheter. AB - A hydrothorax following ventriculoperitoneal shunt catheter insertion is very rare and usually reported in children. Only about 25 cases have been described in the literature and very few have been adults. We report a 51-year-old woman with a massive hydrothorax and respiratory distress following both supradiaphragmatic and transdiaphragmatic migration of a ventriculoperitoneal shunt catheter into the pleural space. To our knowledge this is the first report of the simultaneous occurrence of two types of such migration in one patient. PMID- 17406045 TI - A Chinese family with familial paraganglioma syndrome due to succinate dehydrogenase deficiency. AB - We report the genetic characteristics of a family with familial paraganglioma syndrome. The index patient was diagnosed with carcinoid tumour of the bronchus at the age of 30 years then later diagnosed with bilateral phaeochromocytoma. His sister had bilateral carotid body tumours. Mutational analyses of succinate dehydrogenase B and SDHD on the index patient showed him to be heterozygous for the M1I mutation of the SDHD gene. A genetic analysis revealed that his sister also had succinate dehydrogenase deficiency with the same mutation. Pre symptomatic testing confirmed the genetic diagnosis, and led to a clinical diagnosis in an otherwise asymptomatic sibling. Comparison with other known cases of M1I mutation suggests that this is a founder mutation in the Chinese population. Genetic analysis of the succinate dehydrogenase genes can provide a specific diagnosis and allow for genetic screening of at-risk individuals. PMID- 17406046 TI - Acyclovir-induced nephrotoxicity in a patient with acute retinal necrosis. AB - A 50-year-old woman developed rapidly progressive acute renal failure on the first day after the administration of intravenous acyclovir for acute left retinal necrosis. Intravenous acyclovir was stopped and replaced with intravitreal injections of ganciclovir sodium (2 mg/0.05 mL) and foscarnet (1.2 mg/0.05 mL) 3 times per week for 4 weeks. Acyclovir-induced renal impairment can be reversed if recognised early and treated with careful, timely body fluid replacement. The necrotising retinitis responded well to intravitreal antiviral agents. No complications were seen at the 6 months' follow-up. Constant vigilance is essential for avoiding acute renal failure when treating ophthalmic conditions with intravenous acyclovir. Systematic monitoring of renal function, urine output, and characteristic symptoms like loin pain is warranted. PMID- 17406047 TI - A recalcitrant case of cicatricial pemphigoid. AB - A 57-year-old woman presented with a 1-year history of blisters and erosions on her oral mucosa with bilateral conjunctivitis and symblephara formation. A diagnosis of cicatricial pemphigoid was made based on the clinical features and immunohistological findings. A multidisciplinary team managed her with different topical and systemic immunosuppressive agents but she finally succumbed due to multi-organ failure secondary to sepsis. PMID- 17406048 TI - A man with painless haematuria and hypertension. PMID- 17406049 TI - A woman with a peri-orbital mass. PMID- 17406050 TI - Positive non-sentinel lymph nodes in breast cancer. PMID- 17406051 TI - Diagnosing gastro-intestinal tuberculosis: is the issue acid-fast bacilli? PMID- 17406052 TI - False-negatives in thyroid nodule aspiration cytology. PMID- 17406053 TI - Fine-needle aspiration cytology of the thyroid: screening or diagnostic test? PMID- 17406054 TI - Peritoneography in the diagnosis of peritoneal leaks in peritoneal dialysis. PMID- 17406055 TI - Role of receptor and nonreceptor protein tyrosine kinases in H2O2-induced PKB and ERK1/2 signaling. AB - Excessive generation of reactive oxygen species (ROS) has been implicated in the pathogenesis of many diseases, including atherosclerosis, hypertension, and vascular complications of diabetes. However, the precise mechanisms by which ROS contribute to the development of these diseases are not fully characterized. Hydrogen peroxide (H2O2), a ROS, has been shown to activate several signaling protein kinases, such as extracellular signal-regulated kinase (ERK)1/2 and protein kinase B (PKB) in different cell types, notably in vascular smooth muscle cells. Because these pathways regulate cellular mitogenesis, migration, proliferation, survival, and death responses, their aberrant activation has been suggested to be a potential mechanism of ROS-induced pathologies. The upstream elements responsible for H2O2-induced ERK1/2 and PKB activation remain poorly characterized, but a potential role of receptor and nonreceptor protein tyrosine kinases (PTKs) as triggers that initiate such events has been postulated. Therefore, the aim of this review is to highlight the involvement of receptor and nonreceptor PTKs in modulating H2O2-induced ERK1/2 and PKB signaling. PMID- 17406056 TI - Bax deficiency reduces infarct size and improves long-term function after myocardial infarction. AB - We have previously found that, following myocardial ischemia/reperfusion injury, isolated hearts from bax gene knockout mice [Bax(-/-)] exhibited higher cardioprotection than the wild-type. We here explore the effect of Bax(-/-), following myocardial infarction (MI) in vivo. Homozygotic Bax(-/-) and matched wild-type were studied. Mice underwent surgical ligation of the left anterior descending coronary artery (LAD). The progressive increase in left-ventricular end diastolic diameter, end systolic diameter, in Bax(-/-) was significantly smaller than in Bax(+/+) at 28 d following MI (p < 0.03) as seen by echocardiography. Concomitantly, fractional shortening was higher (35 +/- 4.1% and 27 +/- 2.5%, p < 0.001) and infarct size was smaller in Bax(-/-) compared to the wild-type at 28 days following MI (24 +/- 3.7 % and 37 +/- 3.3%, p < 0.001). Creatine kinase and lactate dehydrogenase release in serum were lower in Bax(-/-) than in Bax(+/+) 24 h following MI. Caspase 3 activity was elevated at 2 h after MI only in the wild-type, but reduced to baseline values at 1 and 28 d post-MI. Bax knockout mice hearts demonstrated reduced infarct size and improved myocardial function following permanent coronary artery occlusion. The Bax gene appears to play a significant role in the post-MI response that should be further investigated. PMID- 17406057 TI - Differential Signaling and Hypertrophic Responses in Cyclically Stretched vs Endothelin-1 Stimulated Neonatal Rat Cardiomyocytes. AB - Numerous neurohumoral factors such as endothelin (ET)-1 and angiotensin (Ang) II as well as the stretch stimulus act concertedly in the in vivo overloaded heart in inducing hypertrophy and failure. The primary culture of rat neonatal cardiomyocytes is the only in vitro model that allows the comparative analysis of growth responses and signaling events in response to different stimuli. In the present study, we examined stretched rat cardiomyocytes grown on flexible bottomed culture plates for hypertrophic growth responses (protein synthesis, protein/DNA ratio, and cell volume), F-actin filaments rearrangement (by confocal laser scanning microscopy), and for signaling events (activation of phospholipase C [PLC]-beta, protein kinase C [PKC], mitogenactivated protein [MAP] kinases) and compared these responses with ET-1 (10-8 M)-stimulated cells. Cyclic stretch for 48 h induced hypertrophic growth in cardiomyocytes indicated by increases in the rate of protein synthesis, cell volume, and diameter, which were less pronounced in comparison to stimulation by ET-1. During cyclic stretch, we observed disoriented F-actin, particularly stress-fibers whereas during ET-1 stimulation, Factins rearranged clearly in alignment with sarcomeres and fibers. The upstream part of signaling by cyclic stretch did not follow the PLCbeta-PKC cascade, which, in contrast, was strongly activated during ET-1 stimulation. Cyclic stretch and, to greater extent, ET-1 stimulated downstream signaling through ERK, p38 MAP kinase, and JNK pathways, but the involvement of tyrosine kinase and PI3 kinase-Akt signaling during cyclic stretch could not be proven. Taken together, our results demonstrate that both cyclic stretch and ET-1 induce hypertrophic responses in cardiomyocytes with different effects on organization of F-actin stress fibers in case of stretch. Furthermore, on the short-term basis, cyclical stretch, unlike ET-1, mediates its hypertrophic response not through activation of PLC-beta and PKC but more likely through integrin-linked pathways, which both lead to downstream activation of the MAP kinase family. PMID- 17406058 TI - Functions of serotonin in hypoxic pulmonary vascular remodeling. AB - In lung vasculature, reversible constriction of smooth muscle cells exists in response to acute decrease in oxygen levels (hypoxia). Progressive and irreversible structural remodeling that reduces blood vessel lumen takes place in response to chronic hypoxia and results in pulmonary hypertension. Several studies have shown a role of serotonin in regulating acute and chronic hypoxic responses. In this review the contribution of serotonin, its receptors and transporter in lung hypoxic responses is discussed. Hypoxic conditions modify plasma levels of serotonin, serotonin transporter activity, and expression of 5 HT1B and 5-HT2B receptors. These appear to be required for pulmonary vascular cell proliferation, which depends on the ratio between reactive oxygen species and nitric oxide. A heterozygous mutation was identified in the 5-HT2B receptor gene of a patient who developed pulmonary hypertension after fenfluramines anorexigen treatment. This C-terminus truncated 5-HT2B mutant receptor presents lower nitric oxide coupling, and higher cell proliferation capacity than the wild type receptor. Under low oxygen tension, cells increase the transcription of specific genes via stabilization of the transcription factor hypoxia-inducible factor (HIF)-1. Factors such as angiotensin II or thrombin that can also control HIF-1 pathway contribute to pulmonary vascular remodeling. The 5-HT2B receptor via phosphatidylinositol-3 kinase/Akt activates nuclear factor-kappaB, which is involved in the regulation of HIF-1 expression. Acontrol of HIF- 1 by 5-HT2B receptors explains why expression of pulmonary vascular remodeling factors, such as endothelin-1 or transforming growth factor-beta, which is HIF-1-alpha regulated, is not modified in hypoxic 5-HT2B receptor mutant mice. Understanding the detailed mechanisms involved in lung hypoxic responses may provide general insight into pulmonary hypertension pathogenesis. PMID- 17406059 TI - Influence of hyperthyroidism on the effect of adenosine transport blockade assessed by a novel method in guinea pig atria. AB - The aim of the present study was to investigate the effect of hyperthyroidism on the trans-sarcolemmal adenosine (Ado) flux via equilibrative and nitrobenzylthioinosine (NBTI)-sensitive nucleoside transporters (ENT1) in guinea pig atria, by assessing the change in the Ado concentration of the interstitial fluid ([Ado]ISF) under nucleoside transport blockade with NBTI. For the assessment, we applied our novel method, which estimates the change in [Ado]ISF utilizing the altered inotropic response to N6-cyclopentyladenosine (CPA), a relative stable selective agonist of A1 Ado receptors, by providing a relative index, the equivalent concentration of CPA. Our results show an interstitial Ado accumulation upon ENT1 blockade, which was more extensive in the hyperthyroid samples (CPA concentrations equieffective with the surplus [Ado]ISF were two to three times higher in hyperthyroid atria than in euthyroid ones, with regard to the negative inotropic effect of CPA and Ado). This suggests an enhanced Ado influx via ENT1 in hyperthyroid atria. It is concluded that hyperthyroidism does not alter the prevailing direction of the Ado transport, moreover intensifies the Ado influx in the guinea pig atrium. PMID- 17406060 TI - Endocardial endothelial cells stimulate proliferation and collagen synthesis of cardiac fibroblasts. AB - Given that vascular endothelial cells play an important role in the modulation of vascular structure and function, we hypothesized that endocardial endothelial cells (EECs) may have a modulator role in regulating the cardiac interstitial cells. Endocardial endothelial cells were isolated from freshly collected pig hearts and cardiac fibroblasts were isolated from 3- to 4-d-old Wistar rats. Fibroblasts were cultured in the presence or absence of conditioned medium from EECs. Proliferation of cardiac fibroblasts was measured by the incorporation of [3H]- Thymidine and collagen synthesis was assayed by the incorporation of [3H] Proline. To determine the involvement of signaling mediators, in separate experiments, cardiac fibroblasts were incubated with BQ123 (selective ETA receptor antagonist), PD142893 (nonselective ETA/ETB receptor antagonist), Bis indolylmaleimide (PKC inhibitor), PD 098059 (MEK inhibitor), or neutralizing anti transforming growth factor (TGF)-beta-antibody. Endocardial endothelium-derived factors endothelin (ET)-1, TGF-beta, and Angiotensin (Ang)-II in the conditioned medium were assayed by enzyme-linked immunosorbent assay using commercially available kits. We report here evidence that suggest that endocardial endothelial cells stimulate both proliferation and collagen synthesis of cardiac fibroblasts. The response seems to be mediated by endothelin through its ETA receptor. Our results also indicate that protein kinase C (PKC) and mitogen-activated protein kinase (MAPK) pathways are essential for the EEC-induced proliferation of cardiac fibroblasts. PMID- 17406061 TI - Expression of signaling molecules associated with apoptosis in human ischemic stroke tissue. AB - There is growing evidence that, because of the highly significant differences in gene activation/protein expression between animal models of stroke and stroke patients, the current treatment strategies based on animal stroke models have been unsuccessful. Therefore, it is imperative that the pathobiology of human stroke be studied. As a first step here, Western blotting and immunohistochemistry were employed to examine expression and tissue localization of key apoptotic proteins in infarct and peri-infarcted (penumbra) from grey and white matter in human postmortem tissue of 18 patients who died between 2 and 37 d after stroke caused by large vessel disease. The contralateral hemisphere was used as a control. JNK1, JNK2, and p53 were upregulated in the majority of samples, whereas Bcl-2, caspase-3, active caspase-3, phosphorylated p53 (p-p53), phosphorylated JNK1 (p-JNK1), and phosphorylated JNK2 (p-JNK2) were upregulated in approximately half of the samples. JNK1 expression was positively correlated with JNK2 expression in grey and white matter infarct and penumbra, whereas active caspase-3 levels were positively correlated with p-JNK2 levels in grey and white matter infarct. Using indirect immunoperoxidase staining of paraffin embedded sections, active caspase-3 was found in infarcted neurons that co localized with TUNEL-positive cells. p-JNK localization in the nuclei of TUNELpositive cells with the morphological appearance of neurons from infarct and penumbra was also demonstrated. The use of Kaplan Meier survival data demonstrated that the presence of Bcl-2 in penumbra of grey matter correlated significantly with shorter survival (p = 0.006). In conclusion, the present study has identified significantly altered expression of apoptotic proteins in human stroke tissue and shown that the presence of Bcl-2 in penumbra of grey matter has prognostic value. It is tempting to suggest that further studies of apoptotic proteins in human stroke may lead to identification of novel targets for drug discovery. PMID- 17406062 TI - Prevention of hypertension in DOCA-salt rats by an inhibitor of soluble epoxide hydrolase. AB - Cyclooxygenase and lipoxygenase metabolism of arachidonic acid produces compounds important in cardiovascular control. Further, arachidonic acid can be metabolised by cytochrome p450 to produce epoxyeicosatrienoic acids (EETs). These derivatives are inactivated by soluble epoxide hydrolase (sEH). The potential role of these EETs in hypertension and cardiac remodelling has been determined using the selective sEH inhibitor, N-adamantyl-N'-dodecylurea (ADU), in deoxycorticosterone acetate (DOCA)-salt hypertensive rats. Experiments were performed on male Wistar rats following uninephrectomy alone (UNX rats) or uninephrectomy with administration of DOCA (25 mg every fourth day subcutaneously) and 1% NaCl in drinking water (DOCA-salt rats). ADU (10 mg/kg/d subcutaneously) was administered for 2 wk starting 2 wk after surgery. Cardiovascular structure and function were determined using organ wet weights, histological analysis of collagen and inflammation, isolated heart and thoracic aortic ring preparations, and electrophysiological measurements. DOCA-salt hypertensive rats developed hypertension, hypertrophy, perivascular and interstitial fibrosis, endothelial dysfunction, and prolongation of the cardiac action potential duration within 4 wk. Administration of ADU prevented the further increase in systolic blood pressure and left-ventricular wet weight and normalized endothelial function. ADU treatment did not change inflammatory cell infiltration, collagen deposition, or cardiac action potential duration. EETs may be involved in the development of hypertension and endothelial dysfunction in DOCA-salt rats, but not in excessive collagen deposition or electrophysiological abnormalities. PMID- 17406063 TI - Cyclic GMP modulates the expression of Gi protein and adenylyl cyclase signaling in vascular smooth muscle cells. AB - We have recently shown that the nitric oxide (NO) donor, SNAP, decreased the expression of Gialpha proteins and associated functions in vascular smooth muscle cells. Because NO stimulates soluble guanylyl cyclase and increases the levels of guanosine 3?',5?'-cyclic monophosphate (cGMP), the present studies were undertaken to investigate whether cGMP can also modulate the expression of Gi proteins and associated adenylyl cyclase signaling. A10 vascular smooth muscle cells (VSMCs) and primary cultured cells from aorta of Sprague Dawley rats were used for these studies. The cells were treated with 8-bromoguanosine 3?',5?' cyclic monophosphate (8BrcGMP) for 24 h and the expression of Gialpha proteins was determined by immunobloting techniques. Adenylyl cyclase activity was determined by measuring [32P]cAMP formation for [alpha-32P]ATP. Treatment of cells with 8-BrcGMP (0.5 mM) decreased the expression of Gialpha-2 and Gialpha-3 by about 30-45%, which was restored towards control levels by KT5823, an inhibitor of protein kinase G. On the other hand, the levels of Gsalpha protein were not altered by this treatment. The decreased expression of Gialpha proteins by 8Br-cGMP treatment was reflected in decreased Gi functions. For example, the inhibition of forskolin (FSK)-stimulated adenylyl cyclase activity by low concentrations of GTPgammaS (receptor-independent Gi functions) was significantly decreased by 8Br-cGMP treatment. In addition, exposure of the cells to 8Br-cGMP also resulted in the attenuation of angiotensin (Ang) II- and C-ANP4-23 (a ring deleted analog of atrial natriuretic peptide [ANP])-mediated inhibition of adenylyl cyclase activity (receptor-dependant functions of Gi). On the other hand, Gsalpha-mediated stimulations of adenylyl cyclase by GTPgammaS, isoproterenol and FSK were significantly augmented in 8Br-cGMP-treated cells. These results indicate that 8Br-cGMP decreased the expression of Gialpha proteins and associated functions in VSMCs. From these studies, it can be suggested that 8Br-cGMP-induced decreased levels of Gi proteins and resultant increased levels of cAMP may be an additional mechanism through which cGMP regulates vascular tone and thereby blood pressure. PMID- 17406064 TI - Vasoactive peptides upregulate mRNA expression and secretion of vascular endothelial growth factor in human airway smooth muscle cells. AB - Airway remodeling and associated angiogenesis are documented features of asthma, of which the molecular mechanisms are not fully understood. Angiotensin (ANG)II and endothelin (ET)-1 are potent vasoconstricting circulatory hormones implicated in asthma. We investigated the effects of ANG II and ET-1 on human airway smooth muscle (ASM) cells proliferation and growth and examined the mRNA expression and release of the angiogenic peptide, vascular endothelial growth factor (VEGF). Serum deprived (48 h) human ASM cells were incubated with ANG II (100 nM) or ET-1 (10 nM) for 30 min, 1, 2, 4, 8, 16, and 24 h and the endogenous synthesis of VEGF was examined in relation to control cells receiving serum free culture medium. ET 1 induced time dependent DNA biosynthesis as determined by [3H]-thymidine incorporation assay. Using northern blot hybridization, we detected two mRNA species of 3.9 and 1.7 kb encoding VEGF in the cultured smooth muscle cells. Both ANG II and ET-1 induced the mRNA expression (two- to threefold) and secretion (1.8- to 2.8-fold) of VEGF reaching maximal levels between 4-8 h of incubation. Induced expression and release of VEGF declined after 8 h of ANG II incubation while levels remained elevated in the case of ET-1. The conditioned medium derived from ET-1-treated ASM cells induced [3H]-thymidine incorporation and cell number in porcine pulmonary artery endothelial as well as human umbilical vein endothelial cells. Moreover, the VEGF tyrosine kinase receptor inhibitor blocked the conditioned medium induced mitogenesis in endothelial cells. Our results suggest a potential role for ANG II and ET-1 in ASM cell growth and upregulation of VEGF that may participate in endothelial cell proliferation via paracrine mechanisms and thus causing pathological angiogenesis and vascular remodelling seen during asthma. PMID- 17406065 TI - Functional polymorphism and differential regulation of CYSLTR1 transcription in human airway smooth muscle and monocytes. AB - Cysteinyl leukotrienes play an important role in the pathophysiology of many inflammatory disorders, including asthma. The aim of this study was to characterize the mechanisms underlying transcriptional regulation of the human cysteinyl leukotriene receptor 1 (hCYSLTR1) gene. 5'RACE was performed on human airway smooth muscle (HASM) and peripheral blood mononuclear cells. A1128-bp region of the hCYSLTR1 main putative promoter was screened for polymorphisms by sequencing of 48 individuals. Luciferase reporter gene assays were performed using fragments of the core promoter (232 bp to 1128 bp) in HASM and THP1 cells. Three hCYSLTR1 transcripts were found, one representing 90% of all messenger RNA identified. The genomic location of the transcription start sites suggested there are two putative hCYSLTR1 promoters. The majority of the transcriptional activity of the main putative promoter was detected between -232 and -679 bp. Four singlenucleotide polymorphisms in strong linkage disequilibrium were found in the region studied: -561 (rs7066737), -642 (rs2806489), -781 (rs2637204), and -940 (rs321029), with three haplotypes observed. In THP1 cells, the G allele (-642) caused a twofold decrease in luciferase expression compared to the Aallele. These data suggest that the majority of hCYSLTR1 transcripts in HASM and monocytes arise from a single promoter located immediately upstream of the 5?' untranslated region, although rarer transcripts can also occur. This study also raises the possibility that cell-type-dependent differences in transcriptional activity caused by the presence of specific haplotypes within the main CYSLTR1 promoter may be a predictor of disease risk or treatment response. PMID- 17406066 TI - Molecular mechanisms in chronic obstructive pulmonary disease: potential targets for therapy. AB - Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease associated with progressive airflow obstruction. Tobacco smoking is the main risk factor worldwide. In contrast to asthma, antiinflammatory therapies are rather ineffective in improving chronic symptoms and reducing inflammation, lung function decline, and airway remodeling. Specific drugs that are directed against the remodeling and chronic inflammation, thereby preventing lung tissue damage and progressive lung function decline, must be developed. Experimental models and expression studies suggest that anti-vascular endothelial growth factor (VEGF) receptor strategies may be of use in patients with emphysema, whereas anti-HER1 directed strategies may be more useful in patients with pulmonary mucus hypersecretion, as seen in chronic bronchitis and asthma. Growth factors and cytokines including VEGF, fibroblast growth factors, transforming growth factor beta, tumor necrosis factor-alpha, CXCL1, CXCL8, and CCL2, and signal transduction proteins such as mitogen-activated protein kinase p38 and nuclear factor-kappaB, seem to be important pathogenetic molecules in COPD. Specific antagonists for these proteins may be effective for different inflammatory diseases. However, their efficacy for COPD therapy has not yet been demonstrated. Finally, other drugs such as retinoic acids may provide restoration of lung tissue structure. Such approaches, however, must await the first results of growth factor or cytokine antagonist therapy in chronic lung diseases. PMID- 17406067 TI - Cloning, expression, and promoter analysis of hepatic lipase derived from human hyperplastic adrenals: evidence for alternative mRNA splicing. AB - Human adrenals contain hepatic lipase (HL) activity, which is thought to facilitate the uptake of plasma cholesterol used in steroidogenesis. We show here that full-length HL mRNA is expressed in hyperplastic adrenals of patients with Cushing's disease. In addition, a splice variant that lacks exon-3 was detected in the human adrenals and hepatoma (HepG2) cells, but not in liver. In CAT reporter assays using human NCI-H295R adrenocortical cells, the HL(-685/+13) promoter region was transcriptionally active, and its activity was enhanced twofold by cAMP. In rat adrenals, the HL gene is exclusively transcribed from an alternative promoter within intron-2, resulting in a variant mRNA that lacks exons 1 and 2. By reverse-transcription PCR, we found no evidence for expression of such a variant mRNA in human adrenals, liver, or HepG2 cells. The presence of both full length mRNA and enzyme activity in human adrenals suggests that part of the HL activity is locally synthesized. PMID- 17406068 TI - Preparation and characterization of novel polymeric microcapsules for live cell encapsulation and therapy. AB - This article describes the preparation and in vitro characterization of novel genipin cross-linked alginate-chitosan (GCAC) microcapsules that have potential for live cell therapy applications. This microcapsule system, consisting of an alginate core with a covalently cross-linked chitosan membrane, was formed via ionotropic gelation between calcium ions and alginate, followed by chitosan coating by polyelectrolyte complexation and covalent cross-linking of chitosan by naturally derived genipin. Results showed that, using this design concept and the three-step procedure, spherical GCAC microcapsules with improved membrane strength, suppressed capsular swelling, and suitable permeability can be prepared. The suitability of this novel membrane formulation for live cell encapsulation was evaluated, using bacterial Lactobacillus plantarum 80 (pCBH1) (LP80) and mammalian HepG2 as model cells. Results showed that capsular integrity and bacterial cell viability were sustained 6 mo postencapsulation, suggesting the feasibility of using this microcapsule formulation for live bacterial cell encapsulation. The metabolic activity of the encapsulated HepG2 was also investigated. Results suggested the potential capacity of this GCAC microcapsule in cell therapy and the control of cell signaling; however, further research is required. PMID- 17406069 TI - Serum copper and zinc concentrations in healthy children aged 3-14 years in Greece. AB - Zinc (Zn) and copper (Cu) are essential trace elements in people, being required for functional activity of several enzyme systems. In this study, we determined Cu and Zn concentrations in the serum of 105 healthy children randomly selected, aged 3-14 yr, residing in a region of Greece (Thrace), and we investigated their association with children's gender, age, height, weight, and nutritional habits. The mean levels of Zn and Cu were 15.01 +/- 2.95 micromol/L and 26.18 +/- 5.47 micromol/L, respectively, with no significant difference between boys and girls. Asignificant positive correlation was found between age and Zn levels, and a negative one was found between age and Cu levels. Both Zn and Cu levels tended to increase with height, whereas Zn levels significantly decreased with increasing body mass indent (BMI). The consumption of meat, milk, and eggs were independent determinants for higher Zn levels, and the consumption of legumes and fruits were independent determinants for higher Cu levels. Asignificant negative correlation was found between Zn and Cu levels. In conclusion, our study, the first one evaluating the serum status of Cu and Zn in healthy Greek children, identified significant correlations of Zn and Cu levels with their age, height, BMI, and nutritional habits. PMID- 17406070 TI - Weak relationship between ionized and total magnesium in serum of patients requiring magnesium status. AB - Measurement and monitoring of magnesium (Mg) are important to prevent the development of serious and potentially fatal complications in critically ill patients. Although ion-selective electrodes are available and earlier reports suggest that free ionized magnesium (iMg2+) is the most useful test to estimate Mg status, most clinical laboratories still only measure total Mg. To compare the relationship among iMg2+, total Mg, and albumin in serum, samples were collected from 48 consecutive patients admitted to an intensive care unit or a primary health center. The mean serum level of iMg2+ in 44 patients was 0.53 mmol/L, the total Mg was 0.96 mmol/L, and the albumin was 34.93 g/L. The correlation between iMg2+ and total Mg in serum was r=0.585; the correlation between iMg2+ and albumin in serum was r=378; and the correlation between total Mg and albumin in serum was r=0.340. The mean percent iMg2+ in relation to total Mg in serum was calculated to be 55% in the patient samples. The important level of biologically active iMg2+ was not reflected upon analysis of total Mg in 25% of consecutive patients. This report shows that the correlation of iMg2+ and total Mg is weak, not only in critically ill patients but also in patients in whom Mg status is inquired as a whole. PMID- 17406071 TI - Plasma zinc, copper, leptin, and body composition are associated in elite female judo athletes. AB - This study evaluated levels of plasma zinc, copper, and leptin, body composition, and their relationship in nine elite female judo athletes under two different training conditions. Body composition and biochemical measurements (hematological indexes, plasma zinc, plasma copper, and plasma leptin) were analyzed 24 h after intense training and following a 5-d period without training (no-training). Plasma leptin and plasma zinc increased with no-training. Plasma zinc correlated negatively with percent fat mass (r= -0.62; r=0.05) and positively with plasma leptin (r=0.83; p=0.002) in the no-training condition. Plasma copper did not change during the study and correlated positively with plasma leptin (r=0.66; p=0.05) and with percent fat mass (r=0.80; p=0.007) after training. Percent fat mass was associated negatively with plasma zinc (r=-0.62; p=0.05) in the no training condition. Moreover, percent fat mass was negatively associated with the Zn/Cu plasma ratio under both training conditions (r< -0.78, p< 0.001). These results are consistent with the possible function of zinc as a lipid-mobilizing factor and of copper as a limiting factor in energy metabolism. PMID- 17406072 TI - Study of iron homeostasis following partial hepatectomy in rats with chronic aluminum intoxication. AB - Effects of both chronic aluminum (Al) exposure and partial hepatectomy on iron (Fe) homeostasis were studied. Male Wistar rats were intraperitoneally administered either 27 mg Al/kg body weight (as aluminum hydroxide) or the vehicle saline, three times a week for 3 mo. After this time, half of the rats of each group was sham operated (SH) and the other half was partially hepatectomized (PH). Animals of the four experimental groups (vehicle+SH [SH]; Al+SH; vehicle+PH [PH], and Al+PH) were killed 48 h after the surgical procedure. Serum, hepatic, and intestinal Al levels were found to be increased both for Al+SH and Al+PH. The serum Fe concentration and transferrin saturation percentage were significantly diminished in the rats of the Al+PH group, thus showing interaction between Al administration and PH. The 59Fe mucosal-to-serosal transport, studied in the intestinal loop in situ, was not affected by Al or PH. The malregulation of intestinal Fe absorption in Al exposure and/or PH when the serum Fe concentration was diminished could be the result of the increased lipid peroxidation (thiobarbituric acid-reactive substances [TBARS]) observed in this tissue. Mucosal TBARS were increased by Al exposure (+26%) and PH (+37%) and interaction between Al and PH was observed (+44%). These results show that when liver surgery is performed after prolonged Al exposure, it leads to impairment of Fe homeostasis. We underline the importance of the exposure to Al, a potentially toxic element, in the study of risk assessment in patients who must be submitted to major liver resection. PMID- 17406073 TI - Influence of aqueous extract of Hibiscus sabdariffa L. petal on cadmium toxicity in rats. AB - The effects of chronic exposure to cadmium (Cd) on some selected biochemical parameters, as well as the possible protective role of aqueous extracts of Hibiscus sabdariffa L petal were studied in 12-wk-old male Wistar albino rats. Exposure to Cd caused a significant increase in plasma Lalanine aminotransferases (ALT) only but with a corresponding decrease in liver L-alanine and L-aspartate aminotransferases (L-ALT, L-AST) when compared to the Cd-free control. Total superoxide dismutase activity was decreased in the liver, testis, and prostate of Cd-exposed rats, whereas malondialdehyde (MDA) concentrations were increased relative to the Cd-free control. The metal significantly increased prostatic acid phosphatase activity in the prostate, but decreased the body weight gain of the rats and organ/body weight ratio for prostate and testis compared to the Cd-free control. Pretreatment of rats with aqueous extract of H. sabdariffa resulted in significantly less hepatotoxicity than with Cd alone as measured by plasma ALT and liver ALT and AST activities. The extract also protected the rats against Cd induced liver, prostate, and testis lipoperoxidation as evidenced by significantly reduced MDA values in these organs, as well as reduced prostatic acid phosphatase activity in the prostate, when compared to the Cd-only exposed rats. Also, when compared to the organ/body weight ratios obtained from rats exposed to Cd alone the prostate and testis were protected by the extract as shown by enhanced prostate/body weight and testis/body weight ratios of Cd- and extract-treated rats. These data suggest that H. sabdarrifa L might be protective in Cd toxicity. PMID- 17406074 TI - Effect of an inhibitor of nitric oxide production on Cu-Zn/SOD and its cofactors in diabetic rats. AB - The aim of this study was to investigate the effect of an inhibitor of nitric oxide production, N(omega)-nitro-L-arginine methyl esther (L-NAME) on Cu-Zn/SOD (superoxide dismutase) enzyme activity and copper and zinc concentrations in diabetes-induced rats. The control group consisted of 12 male albino Sprague Dawley rats, 10-12 wk of age and weighing 300 g. Twenty-six albino Sprague-Dawley rats, 10-12 wk of age and weighing 315 g, constituted the experimental group. The experimental group was divided into two groups. The first group (n=12) constituted streptozotocininduced (55 mg/kg, intraperitoneally) diabetic rats and the second group (n=14) was administered L-NAME (1 mg/kg/d) after streptozotocin induction. For determination of Cu-Zn/SOD activity, spectrophotometry was used. Zinc and copper concentrations were determined by atomic absorption spectrophotometry. Results showed that Cu-Zn/SOD activity was increased significantly in both experimental groups compared to controls, and the increase in the second group was higher than in the first group (p<0.01, p<0.01, p<0.05). Plazma zinc concentration was increased in the second group when compared with controls (p<0.05). Plasma copper was decreased significantly in the second group compared to controls and the first group (p<0.001, p<0.001). Red cell copper concentration was decreased significantly in the first group compared to controls (p<0.05). This study showed that L-NAME administration has ensured an additive effect on the antioxidant defense system, which was proved by the increase in Cu Zn/SOD activity. This increase might have a protective effect against tissue damage in the acute period, with corresponding changes in zinc and copper concentrations. PMID- 17406075 TI - Nickel localization on tissues of hyperaccumulator species of phyllanthus L. (Euphorbiaceae) from ultramafic areas of Cuba. AB - Two species of perennial Phyllanthus (Euphorbiaceae) (Phyllanthus orbicularis and Phyllanthus discolor, both endemic to ultramafic areas of Cuba, and their natural hybrid, Phyllanthus xpallidus) were selected for metal localization microanalysis. Different plant tissues were analyzed by X-ray fluorescence, inductively coupled plasma-atomic emission spectroscopy, and scanning electron microscopy coupled with an energy-dispersive X-ray probe. All of the studied taxa are nickel (Ni) hyperaccumulators and significant concentrations of this element were found in different leaf and stem tissues. The highest Ni content was found in the laticifer tubes, whereas leaf epidermis Ni content resulted to be much more relevant in terms of total metal storage. Calcium and magnesium were found more evenly distributed in leaf and stem tissues. PMID- 17406076 TI - Rapid and direct determination of selenium, copper, and zinc in blood plasma by flow injection-inductively coupled plasma-mass spectrometry. AB - A flow injection-inductively coupled plasma-mass spectrometry (FIICP- MS) with a simple sample preparation procedure was developed for the determination of selenium, copper, and zinc in blood serum/plasma. A serum/plasma sample was filtered through a 0.45-mum membrane filter and diluted with a mixture of trace elements in a standard solution (9 : 1, v/v). Measurement of the reference serum sample confirmed the accuracy of our method for selenium, copper, and zinc concentration. In the case of blood plasma samples obtained from six healthy adult males, the selenium, copper, and zinc concentrations were similar to those of a typical healthy male in Japan. These results suggest that the sample preparative procedure coupled with FI-ICP-MS can be used for the routine determination of selenium, copper, and zinc in human blood serum/plasma. PMID- 17406078 TI - HIV testing: rationale for changing recommendations. AB - HIV testing is an important and effective strategy for preventing HIV infection. Infected individuals who know their HIV serostatus are less likely to engage in high-risk sexual behavior, and it is estimated that knowledge of HIV serostatus in unaware persons could reduce new infections by more than 30%. The availability of rapid testing for HIV expands testing opportunities. Expanded routine, voluntary, and opt-out screening in health care settings is needed to reduce the number of persons who are unaware of their HIV-infected status, get newly diagnosed patients into care, and reduce transmission of HIV infection. This article summarizes a presentation on revisions to Centers for Disease Control and Prevention HIV screening recommendations made by Robert S. Janssen, MD, at the 9th Annual Ryan White CARE Act Clinical Update in Washington, DC, in August 2006. The original presentation is available as a Webcast at www.iasusa.org. PMID- 17406077 TI - HIV infection in women: perinatal issues and cervical cancer surveillance. AB - Issues of HIV infection in women include perinatal care to prevent mother-child transmission and screening for cervical dysplasia. Antiretroviral therapy has been very successful in reducing perinatal transmission rates. Ongoing issues in this setting include absense of relevant pharmacokinetics data for new drugs and formulations, implementation of new resistance testing guidelines, and recent apparently conflicting findings on the potential role of protease inhibitor treatment in preterm delivery. Recent findings also include a similar low transmission rate with vaginal delivery and emergency cesarean delivery versus elective cesarean delivery in women on antiretroviral therapy with HIV viral load of less than 1000 copies/mL, and a low rate of postpartum morbidity in women undergoing elective cesarean delivery. Recent changes in recommendations for cervical cancer screening in the general population should not be applied to HIV infected women. However, the recent finding that HIV-infected women with CD4+ cell counts greater than 500/ul do not have a greater rate of squamous intraepithelial lesions than women without HIV infection suggests that the former can be followed less frequently if they have normal Pap tests. This article summarizes a presentation on HIV infection in women made by Carmen D. Zorrilla, MD, at the 9th Annual Ryan White CARE Act Clinical Update in Washington, DC, in August 2006. The original presentation is available as a Webcast at www.iasusa.org. PMID- 17406079 TI - Syphilis and other sexually transmitted diseases in HIV infection. AB - An ongoing resurgence of syphilis and continued transmission of other common sexually transmitted diseases (STDs) in HIV-infected patients is fueled by a number of factors, including "prevention burnout" resulting from fatigue with long-term, safer-sex behavior, use of recretional drugs (notably methamphetamine), and false sense of security associated with HIV serosorting and elevated CD4+ cell count. Annual screening for common STDs is recommended for HIV infected patients. Issues in syphilis and herpes simplex virus-2 (HSV-2) diagnosis and treatment are discussed. New problems are briefly reviewed, which include the increased reporting of lymphogranuloma venereum and the increased frequency of fluoroquinolone-resistant gonorrhea. The recently revised Centers for Disease Control and Prevention guidelines for treatment of syphilis, HSV-2 infection, chlamydial infection, and gonorrhea are summarized. This article summarizes a presentation on syphilis and other STDs made by Jeanne Marrazzo, MD, MPH, at the International AIDS Society-USA Continuing Medical Education course in New York, in October 2006. The original presentation is available as a Webcast at www.iasusa.org. PMID- 17406080 TI - Modeling synaptic transmission of the tripartite synapse. AB - The tripartite synapse denotes the junction of a pre- and postsynaptic neuron modulated by a synaptic astrocyte. Enhanced transmission probability and frequency of the postsynaptic current-events are among the significant effects of the astrocyte on the synapse as experimentally characterized by several groups. In this paper we provide a mathematical framework for the relevant synaptic interactions between neurons and astrocytes that can account quantitatively for both the astrocytic effects on the synaptic transmission and the spontaneous postsynaptic events. Inferred from experiments, the model assumes that glutamate released by the astrocytes in response to synaptic activity regulates store operated calcium in the presynaptic terminal. This source of calcium is distinct from voltage-gated calcium influx and accounts for the long timescale of facilitation at the synapse seen in correlation with calcium activity in the astrocytes. Our model predicts the inter-event interval distribution of spontaneous current activity mediated by a synaptic astrocyte and provides an additional insight into a novel mechanism for plasticity in which a low fidelity synapse gets transformed into a high fidelity synapse via astrocytic coupling. PMID- 17406081 TI - Speckled microtubules improve tracking in motor-protein gliding assays. AB - Gliding assays of motor proteins such as kinesin, dynein and myosin are commonly carried out with fluorescently labeled microtubules or filamentous actin. In this paper, we show that speckled microtubules (MTs), prepared by copolymerizing 98% unlabeled tubulin with 2% rhodamine-labeled tubulin, can be localized to +/-7.4 nm (24 measurements) in images acquired every 125 ms. If the speckled MTs move at about 800 nm s(-1), ten images are sufficient to determine their velocity to a precision of +/-6.8 nm s(-1) (6 microtubules, 24 measurements). This velocity precision is four-fold better than manual methods for measuring the gliding velocity of uniformly labeled MTs by end-point localization. The improved velocity precision will permit the determination of velocity-force curves when one, two and three kinesin motors pull a single load in vitro. PMID- 17406082 TI - Reactive boundary conditions for stochastic simulations of reaction-diffusion processes. AB - Many cellular and subcellular biological processes can be described in terms of diffusing and chemically reacting species (e.g. enzymes). Such reaction-diffusion processes can be mathematically modelled using either deterministic partial differential equations or stochastic simulation algorithms. The latter provide a more detailed and precise picture, and several stochastic simulation algorithms have been proposed in recent years. Such models typically give the same description of the reaction-diffusion processes far from the boundary of the simulated domain, but the behaviour close to a reactive boundary (e.g. a membrane with receptors) is unfortunately model-dependent. In this paper, we study four different approaches to stochastic modelling of reaction-diffusion problems and show the correct choice of the boundary condition for each model. The reactive boundary is treated as partially reflective, which means that some molecules hitting the boundary are adsorbed (e.g. bound to the receptor) and some molecules are reflected. The probability that the molecule is adsorbed rather than reflected depends on the reactivity of the boundary (e.g. on the rate constant of the adsorbing chemical reaction and on the number of available receptors), and on the stochastic model used. This dependence is derived for each model. PMID- 17406083 TI - Positive feedback, stochasticity and genetic competence. AB - A single gene, regulating its own expression via a positive feedback loop, constitutes a common motif in gene regulatory networks and signalling cascades. Recent experiments on the development of competence in the bacterial population B. subtilis show that the autoregulatory genetic module by itself can give rise to two types of cellular states. The states correspond to the low and high expression states of the master regulator ComK. The high expression state is attained when the ComK protein level exceeds a threshold value leading to a full activation of the autostimulatory loop. Stochasticity in gene expression drives the transitions between the two stable states. In this paper, we explain the appearance of bimodal protein distributions in B. subtilis cell population in the framework of three possible scenarios. In two of the cases, bistability provides the basis for binary gene expression. In the third case, the system is monostable in a deterministic description and stochasticity in gene expression is solely responsible for the appearance of the two expression states. PMID- 17406084 TI - Surface waves of Min-proteins. AB - In the bacterium Escherichia coli, the Min-proteins show pronounced pole-to-pole oscillations. They are functional for suppressing cell division at the cell ends, leaving the center as the only possible site for division. Analyzing different models of Min-protein dynamics in a bacterial geometry, we find waves on the cytoplasmic membrane. Interestingly, the surface wave solutions of different models belong to different symmetry classes. We suggest that experiments on Min protein surface waves in vitro are helpful in distinguishing between different classes of models of Min-protein dynamics. PMID- 17406085 TI - Propagation of extrinsic perturbation in a negatively auto-regulated pathway. AB - The apparent precision of the output in multi-step biochemical pathways in the face of external and intrinsic perturbations is non-obvious and conceptually difficult. Using a simple three-step negatively auto-regulated model pathway, we show that the effect of perturbation at different steps of the pathway and its transmission through the network is dependent on the context (i.e., the position) of the particular reaction step in relation to the topology of the regulatory network, stoichiometry of reactions, type of nonlinearity involved in the reactions and also on the intrinsic dynamical state of the pathway variables. We delineate the qualitative and quantitative changes in the pathway dynamics for constant ('bias') and random external perturbations acting on the pathway steps locally or globally to all steps. We show that constant perturbation induces qualitative change in dynamics, whereas random fluctuations cause significant quantitative variations in the concentrations of the different variables. Thus, the dynamic response of multi-step biochemical pathways to external perturbation depends on their biochemical, topological and dynamical features. PMID- 17406086 TI - Comment on elastic network models and proteins. AB - Elastic network models have been used to study the properties of coarse grained models of proteins and larger biomolecular complexes. In this comment, we point out that it is important to build rotational symmetry, as well as translational symmetry, into these models that are designed to describe the rigidity, and the associated low-frequency deformations. This leads to strong restrictions on what form of interactions can be used. In particular, the only allowed two-center harmonic interactions are those corresponding to Hooke springs. Additional complexity can be introduced if required by using three-center harmonic interactions. PMID- 17406088 TI - Sexual conflict in plants. PMID- 17406089 TI - Microsatellite DNA typing for assessment of genetic variability in Tharparkar breed of Indian zebu (Bos indicus) cattle, a major breed of Rajasthan. AB - The present study estimates genetic variability with a set of 25 microsatellite markers in a random sample of 50 animals of Tharparkar breed of Indian zebu (Bos indicus) cattle. Tharparkar is a dual-purpose breed, valued for its milk as well as draught utility, and is adapted to the inhospitable Thar desert conditions of Rajasthan typified by summer temperature hovering above 50 degrees C, sparse rainfall and vegetation, and scarcity of even drinking water. The observed number of alleles ranged from 4 (ETH3, ILSTS030, INRA5, INRA63 and MM8) to 11 (HEL9 and ILSTS034), with allelic diversity (average number of observed alleles per locus) of 6.20. Observed and expected heterozygosity ranged from 0.25 (INRA63) to 0.77 (ETH10), and from 0.51 (HEL5 and HAUT27) to 0.88 (HEL9) respectively. Wide range of genetic variability supported the utility of these microsatellite loci in measurement of genetic diversity indices in other Indian cattle breeds too. Various average genetic variability measures, namely allele diversity (6.20), observed heterozygosity (0.57), expected heterozygosity (0.67) and mean polymorphism information content (0.60) values showed substantial within-breed genetic variability in this major breed of Rajasthan, despite accumulated inbreeding as reflected by high average inbreeding coefficient (F(IS) = 0.39). The Tharparkar population has not experienced a bottleneck in the recent past. PMID- 17406090 TI - Cloning, characterization and promoter analysis of common carp hairy/Enhancer-of split-related gene, her6. AB - Some members of hairy/Enhancer-of-split-related gene (HES) family have important effects on axial mesoderm segmentation and the establishment and maintenance of the somite fringe. In fishes, the her6 gene, a member of the HES family, is the homologue of hes1 in mammals and chicken. In this study, the her6 gene and its full-length cDNA from the common carp (Cyprinus carpio) were isolated and characterized. The genomic sequence of common carp her6 is approximately 1.7 kb, with four exons and three introns, and the full-length cDNA of 1314 bp encodes a putative polypeptide of 271 amino acids. To analyse the promoter sequence of common carp her6, sequences of various lengths upstream from the transcription initiation site of her6 were fused to enhanced green fluorescent protein gene (eGFP) and introduced into zebrafish embryos by microinjection to generate transgenic embryos. Our results show that the upstream sequence of 500 bp can direct highly efficient and tissue-specific expression of eGFP in zebrafish embryos, whereas a fragment of 200 bp containing the TATA box and a partial suppressor of hairless paired site sequence (SPS) is not sufficient to drive eGFP expression in zebrafish embryos. PMID- 17406091 TI - Recent integrations of mammalian Hmg retropseudogenes. AB - We propose that select retropseudogenes of the high mobility group nonhistone chromosomal protein genes have recently integrated into mammalian genomes on the basis of the high sequence identity of the copies to the cDNA sequences derived from the original genes. These include the Hmg1 gene family in mice and the Hmgn2 family in humans. We investigated orthologous loci of several strains and species of Mus for presence or absence of apparently young Hmg1 retropseudogenes. Three of four analysed elements were specific to Mus musculus, two of which were not fixed, indicative of recent evolutionary origins. Additionally, we datamined a presumptive subfamily (Hmgz) of mouse Hmg1, but only identified one true element in the GenBank database, which is not consistent with a separate subfamily status. Two of four analysed Hmgn2 retropseudogenes were specific for the human genome, whereas a third was identified in human, chimpanzee and gorilla genomes, and a fourth additionally found in orangutan but absent in African green monkey. Flanking target-site duplications were consistent with LINE integration sites supporting LINE machinery for their mechanism of amplification. The human Hmgn2 retropseudogenes were full length, whereas the mouse Hmg1 elements were either full length or 3'-truncated at specific positions, most plausibly the result of use of alternative polyadenylation sites. The nature of their recent amplification success in relation to other retropseudogenes is unclear, although availability of a large number of transcripts during gametogenesis may be a reason. It is apparent that retropseudogenes continue to shape mammalian genomes, and may provide insight into the process of retrotransposition, as well as offer potential use as phylogenetic markers. PMID- 17406092 TI - Sample size computation for association studies using case-parents design. PMID- 17406093 TI - Computational identification of novel PR-1-type genes in Oryza sativa. PMID- 17406094 TI - Genetic characterization of Zanskari breed of horse. PMID- 17406095 TI - Assessment of genetic diversity of Lycoris longituba (Amaryllidaceae) detected by RAPDs. PMID- 17406096 TI - Reduced larval feeding rate is a strong evolutionary correlate of rapid development in Drosophila melanogaster. PMID- 17406097 TI - The c.IVS1+1G>A mutation in the GJB2 gene is prevalent and large deletions involving the GJB6 gene are not present in the Turkish population. PMID- 17406098 TI - Single-nucleotide polymorphisms in the B7H3 gene are not associated with human autoimmune myasthenia gravis. PMID- 17406099 TI - Accumulation of hns mutations specifically in stationary phase in an E. coli strain carrying an impaired rpoS locus. PMID- 17406100 TI - Abnormal timing of cytokinesis in microsporogenesis in Brachiaria humidicola (Poaceae: Paniceae). PMID- 17406101 TI - Gene action and morphological characteristics of pink flower and pink filament mutants in cotton (Gossypium hirsutum L.). PMID- 17406102 TI - Improvement of the sensitivity and resolution of PCR-SSCP analysis with optimized primer concentrations in PCR products. PMID- 17406103 TI - Antioxidative response mechanisms in halophytes: their role in stress defence. AB - Normal growth and development of plants is greatly dependent on the capacity to overcome environmental stresses. Environmental stress conditions like high salinity, drought, high incident light and low or high temperature cause major crop losses worldwide. A common denominator in all these adverse conditions is the production of reactive oxygen species (ROS) within different cellular compartments of the plant cell. Plants have developed robust mechanisms including enzymatic or nonenzymatic scavenging pathways to counter the deleterious effects of ROS production. There are a number of general reviews on oxidative stress in plants and few on the role of ROS scavengers during stress conditions. Here we review the regulation of antioxidant enzymes during salt stress in halophytes, especially mangroves. We conclude that (i) antioxidant enzymes protect halophytes from deleterious ROS production during salt stress, and (ii) genetic information from mangroves and other halophytes would be helpful in defining the roles of individual isoforms. This information would be critical in using the appropriate genes for oxidative stress defence for genetic engineering of enhanced stress tolerance in crop systems. PMID- 17406104 TI - Cognitive predictors of improvements in adults' spoken word recognition six months after cochlear implant activation. AB - This study investigated whether cognitive measures obtained prior to cochlear implant surgery activation could predict improvements in spoken word recognition in adult cochlear implant recipients 6 months after activation. In addition to noncognitive factors identified by previous studies (i.e. younger age, shorter duration of hearing loss), the present results indicated that improvement in spoken word recognition was associated with higher verbal learning scores and better verbal working memory. Contrary to expectation, neither general cognitive ability nor processing speed was significantly correlated with outcome at 6 months. Multiple regression analyses revealed that a combination of verbal learning scores and lip-reading skill accounted for nearly 72% of the individual differences in improvement in spoken word recognition (i.e. the variance in spoken word recognition scores at 6 months that remained unexplained after controlling for baseline spoken word recognition scores). These findings have relevance for research on auditory processing with cochlear implants as well as implications for clinical interventions. PMID- 17406105 TI - Testing a method for quantifying the output of implantable middle ear hearing devices. AB - This report describes tests of a standard practice for quantifying the performance of implantable middle ear hearing devices (also known as implantable hearing aids). The standard and these tests were initiated by the Food and Drug Administration of the United States Government. The tests involved measurements on two hearing devices, one commercially available and the other home built, that were implanted into ears removed from human cadavers. The tests were conducted to investigate the utility of the practice and its outcome measures: the equivalent ear canal sound pressure transfer function that relates electrically driven middle ear velocities to the equivalent sound pressure needed to produce those velocities, and the maximum effective ear canal sound pressure. The practice calls for measurements in cadaveric ears in order to account for the varied anatomy and function of different human middle ears. PMID- 17406106 TI - Management of complex, surgically intractable intracranial aneurysms: the option for intentional reconstruction of aneurysm neck followed by endovascular coiling. AB - BACKGROUND: A limited series of patients with aneurysm were reviewed retrospectively to analyze strategies for integrating microsurgical and endovascular techniques in the management of complex, surgically intractable aneurysms. METHODS: Four patients were managed in Fujita Health University with a multimodality approach: intentional reconstruction of the aneurysm neck followed by endovascular coiling. RESULTS: A total of 5 aneurysms were treated, of which 3 were large or giant in size, and 3 were fusiform or multilobulated. Complete angiographic obliteration was confirmed in 4 aneurysms (80%). All patients had a good outcome (Glasgow Outcome Scale score 5; mean follow-up, 64 months). CONCLUSION: As for complex, surgically intractable aneurysms, the intentional reconstruction of the aneurysm neck followed by endovascular coiling should be considered more often. PMID- 17406107 TI - Serum asymmetric dimethylarginine negatively correlates with intima-media thickness in early-onset atherosclerosis. AB - BACKGROUND: Asymmetric dimethylarginine (ADMA) assumes a significant role in atherosclerosis by inhibiting the endothelial nitric oxide synthase (eNOS). Moreover, ADMA inhibits the inducible NOS (iNOS), the isoform that triggers atherosclerosis via peroxynitrite formation. Therefore, we investigated whether ADMA is a risk or protective factor in the atherosclerotic process. Intima-media thickness (IMT) of the common carotid artery, a surrogate for vascular diseases, was chosen as the outcome variable of interest. METHODS: Sixty patients younger than 55 years having at least 30% stenosis of the internal carotid artery and 30 age- and gender-matched controls were recruited at a community-based neurosonological laboratory. We investigated relatively young patients to circumvent the confounding effect age has in the development of atherosclerosis. RESULTS: The IMT showed a negative correlation with ADMA upon analysis of the pooled data (Spearman correlation coefficient -0.300, p = 0.0041) and the atherosclerotic stratum (Spearman correlation coefficient -0.323, p = 0.012). A multiple linear regression model containing all determinant factors of IMT previously identified by simple regression was used to further quantify the relationship between IMT and ADMA. The negative association between IMT and ADMA remained statistically significant (beta: -0.510, CI: -0.894, -0.127; p = 0.010), furthermore it was even stronger in the atherosclerotic stratum (beta: -0.67, CI: -1.16, -0.18; p = 0.008). CONCLUSIONS: A minimal increase in ADMA concentration may be protective by inhibiting iNOS but not eNOS in states where iNOS is induced, e.g. inflammation accompanying atherosclerosis. PMID- 17406108 TI - Paraoxonase 2 gene C311S polymorphism is associated with a risk of large vessel disease stroke in a Polish population. AB - BACKGROUND: Oxidative stress plays a role in atherosclerosis. Human paraoxonase (PON) gene products exhibit antioxidant properties. We studied the significance of the Q192R and M55L polymorphisms of the PON1 gene and the C311S polymorphism of the PON2 gene in different etiologies of ischemic stroke. METHODS: One hundred and thirty-six patients with large vessel disease (LVD) stroke, 140 with small vessel disease stroke, 272 with cardioembolic stroke, and their age- and sex matched controls were included. PON genotypes were evaluated by PCR-RFLP analyses. RESULTS: The distribution of PON1 polymorphisms was similar in each stroke group and in the respective controls. Genotypes with the C allele of the PON2 gene C311S polymorphism were overrepresented in LVD stroke patients as compared with their controls, both in univariate and multivariate (dominant model: OR = 1.58, 95% CI: 1.006-2.48) analyses. CONCLUSION: The genotype with the C allele of the PON2 gene is a risk factor for LVD stroke in a Polish population. PMID- 17406109 TI - Coping strategies as determinants of quality of life in stroke patients: a longitudinal study. AB - BACKGROUND: Quality of life (QoL) is reduced for stroke patients and coping strategies have been suggested as determinants of QoL. Thus far the relationship between coping and QoL has only been examined in small-scale cross-sectional designs. Therefore, the current study set out to examine this relationship in a longitudinal setting. METHODS: Stroke patients who were discharged home were interviewed at 4 different time points; just before discharge (T1), and 2 months (T2), 5 months (T3) and 9-12 months after discharge (T4). QoL was measured by the EQ-5D index score and the SF-36 utility score and coping expressed in terms of tenacious goal pursuit and flexible goal adjustment. Modified Rankin scale was assessed as a measure of general functioning. RESULTS: Eighty stroke patients were included. Coping was not predictive of QoL at T1 and T2 but rather at T3 and T4. At T4 both coping strategies determined the levels of QoL as measured with the EQ-5D index score; higher levels of tenacious goal pursuit as well as flexible goal adjustment were associated with higher levels of QoL. This regression model explained 44% of the variance. CONCLUSIONS: Coping is a powerful determinant of QoL, but only more than 5 months after discharge; before this time QoL is mainly determined by general functioning. Both coping strategies were important determinants of QoL. PMID- 17406110 TI - Cognitive disorders in acute stroke: prevalence and clinical determinants. AB - BACKGROUND: Although cognitive impairment early after stroke is a powerful predictor of long-term functional dependence and dementia, little is known about the characteristics and determinants of cognitive dysfunction in acute stroke. METHODS: We administered a neuropsychological examination covering 7 cognitive domains to 190 patients within 3 weeks after a first stroke. We also assembled lesion characteristics, clinical factors at admission, demographic characteristics and vascular risk factors. Multivariate logistic regression adjusted for age, gender and education was performed to examine determinants of acute cognitive impairment. RESULTS: Overall, 74% of patients with a cortical stroke, 46% with a subcortical stroke and 43% with an infratentorial stroke demonstrated acute cognitive impairment. Disorders in executive functioning (39%) and visual perception/construction (38%) were the most common. The prevalence and severity of deficits in executive functioning, language, verbal memory and abstract reasoning was more pronounced following left compared to right cortical stroke (all p < 0.05). Intracerebral haemorrhage (OR = 5.6; 95% CI = 1.2-25.4) and cortical involvement of the stroke (OR = 3.6; 95%, CI = 1.3-9.9) were independent determinants of acute cognitive impairment, whereas premorbid moderate alcohol consumption exerted a protective effect (OR = 0.4; 95% CI = 0.1 1.1). CONCLUSIONS: Cognitive impairment is common in the first weeks after stroke, with executive and perceptual disorders being the most frequent. Intracerebral haemorrhage, cortical involvement of the lesion and premorbid moderate alcohol consumption are independently associated with acute cognitive impairment. PMID- 17406111 TI - Quality of life 6 months after acute stroke: impact of initial treatment in a stroke unit and general medical wards. AB - BACKGROUND AND PURPOSE: Few studies have assessed the influence of the organization of stroke care on health-related quality of life (HRQoL). The present study compared HRQoL 6 months after stroke between subjects who had been treated in an acute stroke unit (SU) and those treated in general medical wards (GMW). METHODS: In total 550 subjects > or = 60 years of age with acute stroke were prospectively allocated according to date of birth (day of the month) to treatment in an SU with short length of stay or GMWs. In the present study, 325 survivors at 5 days with Scandinavian stroke scale (SSS) orientation score > 5 and SSS speech score > 5 were included. We assessed HRQoL with the Short Form 36 (SF-36) questionnaire 6 months after stroke onset and estimated utility values from the SF-36 scores. RESULTS: Of the 325 eligible subjects, 158 were allocated to an SU and 167 to GMWs. Among 296 survivors at 6 months, 115 subjects (79% of survivors) in the SU group completed the questionnaire and 121 (80%) in the GMW group. There was no statistically significant difference between the treatment groups on any of the 8 SF-36 scales or the 2 component summary scales. After integrating death in the comparison of HRQoL, there was still no difference in outcome between the groups. CONCLUSION: An acute SU with a short length of stay, offering early treatment and rehabilitation, could not show an improvement in the HRQoL of stroke patients > or = 60 years 6 months after stroke compared with initial treatment in GMWs. PMID- 17406112 TI - Is transcranial Doppler for the detection of venous-to-arterial circulation shunts reproducible? AB - BACKGROUND: We investigated the reproducibility of contrast transcranial Doppler (TCD), a safe non-invasive test for investigation of venous-to-arterial circulation shunts (v-aCS), usually patent foramen ovale, in young stroke patients. We also investigated whether microbubble contrast was reproducible and whether the addition of blood to agitated saline contrast affected the number of microbubbles produced. METHODS: TCD investigation for v-aCS was repeated in 42 patients using a standardised protocol (i) by the same investigator and (ii) by a different investigator. Agitated saline was produced by mixing saline and 1 ml of air between two 10-ml syringes. The effect of adding blood and increasing the number of agitations was evaluated by microscopy examination using a haemocytometer to assess bubble numbers and sizes. RESULTS: TCD: no difference was found in the highest microbubble count for the same investigator and between different investigators (p > 0.05). Reproducibility for the detection of v-aCS consistent with a patent foramen ovale was also good (kappa values >0.8). Contrast: both the number of contrast mixes before injection and the presence of blood significantly increased the number of bubbles counted. On average, 18 agitations produced 1.86 (95% CI 1.62-2.13) times more bubbles than 6 agitations. Mixtures with blood produced on average 3.8 times more bubbles (3.08-4.69). The size of the bubbles was similar for all mixtures. CONCLUSIONS: Contrast TCD is reproducible and reliable for the detection of v-aCS. The addition of blood and 18 mixes rather than 6 significantly increased the number of microbubbles produced and may increase the effectiveness of microbubble contrast. PMID- 17406113 TI - C-reactive protein level in severe stenosis of cerebral arteries. AB - BACKGROUND: C-reactive protein (CRP) as a non-specific inflammatory biomarker has been demonstrated to actively contribute to all stages of atherogenesis. The aim of this study was to investigate the relation between CRP concentrations and the severity of stenosis of cerebral arteries. METHODS: Lipid parameters and CRP levels were measured in the sera of 119 patients with stenosis of the extracranial cerebral arteries established by angiography and compared with the control group, with a normal appearance of the cerebral arteries on ultrasound examination. CRP concentrations were determined by a high-sensitivity assay. RESULTS: CRP concentrations showed a statistically significant difference in the group of patients with cerebrovascular stenosis of more than 70% compared to the control group (median value 3.4 vs. 1.5 mg/l in the control group, p < 0.05). At logistic regression analysis, CRP was significantly associated with stenosis of more than 70%. CONCLUSION: In addition to traditional biochemical risk markers, an elevated CRP level measured on automated analysers using a high-sensitivity assay may be useful in the detection of patients with severe stenosis of the cerebral arteries. PMID- 17406114 TI - Knowledge and understanding of disease process, risk factors and treatment modalities in patients with a recent TIA or minor ischemic stroke. AB - BACKGROUND: Patients with acute stroke often have a striking lack of knowledge of causes, warning signs, and risk factors. Lack of knowledge may lead to inappropriate secondary prevention behavior. We investigated the knowledge of patients with a TIA or minor stroke about specific aspects of their disease 3 months after the event. METHODS: Patients with a TIA or minor stroke who participated in a randomized controlled trial of the effect of health education by an individualized multimedia computer program (IMCP) were included. All patients received information about their disease from their treating neurologist and half of the patients received extra information through the IMCP. The patients' knowledge was tested after 3 months by means of a questionnaire that contained items on pathogenesis, warning signs, vascular diseases, risk factors, lifestyle and treatment. The highest possible score was 71 points. RESULTS: The 57 patients had a mean total score of 41.2 points (SD 10.4) of the maximum 71. Only 15 (26%) correctly identified the brain as the affected organ in stroke and TIA, and only 21 (37%) could give a correct description of a TIA or stroke. In contrast, 80-90% of the patients identified hypertension and/or obesity as vascular risk factors. Knowledge of various treatment modalities of hypertension, hypercholesterolemia and obesity was moderate to high (40-91% adequate responses). CONCLUSION: The vast majority of patients with TIA or stroke lack specific knowledge about their disease, but they do have a reasonable knowledge of general vascular risk factors and treatment. This suggests that counseling by neurologists of patients with a TIA or stroke can be improved. PMID- 17406115 TI - Duplex scanning diagnosis of internal carotid artery dissections. A case control study. AB - BACKGROUND: The reliability of duplex scanning (DS) for the diagnosis of internal carotid artery dissections (ICAD) is not clear. METHODS: Nine DS signs known to be suggestive for the diagnosis of ICAD were compared between 70 patients with ICAD and 70 matched patients without dissection. RESULTS: Visible internal tapering occlusion, regular eccentric narrowing channel, ectasia beyond the carotid bulb, resistive index asymmetry, blood flow slowdown, ophthalmic artery blood flow inversion, and biphasic flow are more frequent in cases than in controls (p < 0.001). Atheroma plaques were absent in 80% of ICAD. When DS direct signs and hemodynamic signs were studied, sensitivity was 90% and specificity 60%. CONCLUSION: Diagnosis of ICAD by DS could be improved if direct signs were combined with hemodynamic signs, giving a high sensitivity and a rather good specificity. PMID- 17406116 TI - Fibromuscular dysplasia may herald symptomatic recurrence of cervical artery dissection. AB - BACKGROUND: The prevalence of fibromuscular dysplasia (FMD) in patients with cervical artery dissection (CAD) is unknown. Our objectives were to assess the risk of CAD recurring as a stroke or a transient ischemic attack and the association of these events with FMD. METHODS: We prospectively included and followed 103 consecutive patients who had been admitted for a CAD. The median follow-up was 4 years (range 4 months to 10 years). The main criteria for inclusion were a mural hematoma demonstrated by cervical magnetic resonance imaging and/or signs suggesting CAD on 2 other investigations. FMD was diagnosed on the so-called string of beads pattern by digital subtraction angiography. RESULTS: Five patients had CAD recurrence (60% occurred late). Four of these 5 patients had FMD. In 4 patients, CAD recurrence involved another cervical artery. CONCLUSION: The rate of symptomatic CAD recurrence was 1% per year and was often related to FMD. PMID- 17406117 TI - Prognostic markers in gastrointestinal stromal tumors--we are not there yet. PMID- 17406118 TI - The cyclin-dependent kinase inhibitor, p27, has no correlation with the malignant potential of GIST. AB - BACKGROUND: Tumor size and mitotic activity are characteristically associated with the malignant potential and prognosis of gastrointestinal stromal tumors (GIST). However, since neither small tumor size nor low mitotic activity can rule out malignancy, additional factors that may predict malignant behavior have been suggested. AIM: To evaluate the correlation between the cyclin-dependent kinase inhibitor (CDI), p27kip1, expression and the malignant potential of GIST. METHODS: Serial sections were evaluated by immunohistochemistry after staining with antibodies against p27/Kip1 and Ki-67 in surgical material obtained from 36 patients with GIST. p27kip1 staining intensity and the percentage of positive cells were investigated for association with the malignancy risk, pathological features, overall survival, and disease-free survival. Histologic grade was assigned by spindle vs. epithelioid cell histology, mucosal invasion, tumor cell necrosis or atypia and the number of mitoses. RESULTS: In the multivariate model, p27kip1 expression was not significantly associated with any of the variables examined except Kit protein-CD117 (r = 0.37, p = 0.03). Comparative evaluation of p27kip1 expression in GIST cells revealed higher levels of p27kip1 in patients who died compared to those who survived (2.04 +/- 0.54 vs. 1.3 +/- 0.99, p = 0.1). CONCLUSION: The CDI p27kip1 was not associated with malignancy of GISTs and did not serve as a predictor of survival. PMID- 17406119 TI - Small poly-L-lysines improve cationic lipid-mediated gene transfer in vascular cells in vitro and in vivo. AB - The potential of two small poly-L-lysines (sPLLs), low molecular weight sPLL (LMW L) containing 7-30 lysine residues and L18 with 18 lysine repeats, to enhance the efficiency of liposome-mediated gene transfer (GT) with cationic lipid DOCSPER [1,3-dioleoyloxy-2-(N(5)-carbamoyl-spermine)-propane] in vascular smooth muscle cells (SMCs) was investigated. Dynamic light scattering was used for determination of particle size. Confocal microscopy was applied for colocalization studies of sPLLs and plasmid DNA inside cells. GT was performed in proliferating and quiescent primary porcine SMCs in vitro and in vivo in porcine femoral arteries. At low ionic strength, sPLLs formed small complexes with DNA (50-100 nm). At high ionic strength, large complexes (>1 microm) were observed without any significant differences in particle size between lipoplexes (DOCSPER/DNA) and lipopolyplexes (DOCSPER/sPLL/DNA). Both sPLLs were colocalized with DNA inside cells 24 h after transfection, protecting DNA against degradation. DOCSPER/sPLL/DNA formulations enhanced GT in vitro up to 5-fold, in a porcine model using local periadventitial application up to 1.5-fold. Both sPLLs significantly increased liposome-mediated GT. Poly-L-lysine L18 was superior to LMW-L since it enabled maximal GT at a 10-fold lower concentration. Thus, sPLLs may serve as enhancers for GT applications in SMCs in vitro and in vivo using local delivery. PMID- 17406120 TI - Dexamethasone treatment and ICAM-1 deficiency impair VEGF-induced angiogenesis in adult brain. AB - BACKGROUND: Infusion of exogenous vascular endothelial growth factor (VEGF) into adult brain at doses above 60 ng/day induces dramatic angiogenesis accompanied by vascular leak and inflammation. Blood vessels formed by this treatment are dilated and tortuous, exhibiting a pathological morphology. Pathological VEGF induced angiogenesis is preceded by vascular leak and inflammation, which have been proposed to mediate subsequent angiogenesis. METHODS: To test this hypothesis, we infused VEGF into the brains of adult rats to induce pathological angiogenesis. Some of these rats were treated with dexamethasone, a potent anti inflammatory glucocorticoid, to inhibit inflammation and edema. RESULTS: We demonstrate that inhibition of inflammation by treatment with dexamethasone significantly attenuated VEGF-induced pathological angiogenesis. To present converging evidence that inflammation may be important in this angiogenic process, we also demonstrate that mice genetically deficient in the inflammatory mediator intercellular adhesion molecule-1 have attenuated VEGF-induced angiogenesis. These same mice showed normal amounts of physiological angiogenesis in response to enriched environments, however, suggesting that a generalized reduction in vascular plasticity could not account for their poor angiogenic response to VEGF. CONCLUSIONS: Taken together, the data from these experiments suggest that the inflammation which occurs before or during VEGF-induced pathological brain angiogenesis plays a contributory role in the pathological angiogenic process. PMID- 17406121 TI - Membrane cholesterol depletion with beta-cyclodextrin impairs pressure-induced contraction and calcium signalling in isolated skeletal muscle arterioles. AB - OBJECTIVE: Given evidence for clustering of signalling molecules and ion channels in cholesterol-rich membrane domains, the involvement of such structures in arteriolar smooth muscle mechanotransduction was examined. METHOD: To determine the contribution of smooth muscle cholesterol-rich membrane domains to the myogenic response, isolated arterioles were exposed to the cholesterol-depleting agent beta-cyclodextrin (1-10 mM) in the absence and presence of excess exogenous cholesterol. RESULTS: beta-Cyclodextrin significantly impaired pressure-induced vasoconstriction, while excess cholesterol attenuated this effect. Impaired myogenic constriction was evident in de-endothelialized vessels, indicating an action at the level of smooth muscle. beta-Cyclodextrin treatment uncoupled increases in intracellular Ca(2+) from myogenic constriction and depleted intracellular Ca(2+) stores consistent with a loss of connectivity between plasma membrane and sarcoplasmic reticulum signalling. However, beta-cyclodextrin treated arterioles showed unaltered constrictor responses to KCl and phenylephrine. Electron microscopy verified that beta-cyclodextrin caused a decrease in caveolae, while confirmation of smooth muscle containing caveolae was obtained by immunostaining for caveolin-1. Viability of beta-cyclodextrin-treated arterioles was confirmed by agonist sensitivity and propidium iodide nuclear staining. CONCLUSION: The data suggest that smooth muscle cholesterol-rich membrane domains contribute to the myogenic response. Further studies are required to determine whether this relates to specific mechanosensory events or generalized alterations in membrane function. PMID- 17406122 TI - Effect of renal ischemia/reperfusion on gene expression of a pH-sensitive K+ channel. AB - BACKGROUND: Sodium reabsorption depends on the Na/K/ATPase activity coupled to basolateral K+ recycling through K+ channels. ATP depletion reduces pump activity and increases K+ leak resulting in transport dysfunction. Kir4.1 is a pH sensitive K+ channel expressed in the basolateral membrane of distal tubules. In this study, we evaluated whether Kir4.1 is also expressed in proximal tubules (PTs) and whether renal ischemia alters Kir4.1 mRNA expression levels. METHODS: The presence of Kir4.1 mRNA was evaluated in PTs microdissected from collagenase treated rat kidneys. Kir4.1 expression levels were estimated in the renal cortex by multiplex RT-PCR after 30 or 60 min of renal ischemia followed by 1, 24, 48 or 72 h of reperfusion. RESULTS: The PCR product obtained from isolated tubules was sequenced and showed approximately 98% homology with rat Kir4.1 cDNA. Ischemia/reperfusion for 30 min induced a time-dependent reduction in Kir4.1 mRNA expression in parallel with plasma creatinine, however recovery was delayed after 60 min of ischemia, remaining reduced after 72 h of reperfusion when plasma creatinine was already normalized. CONCLUSION: Kir4.1 mRNA expression was decreased by renal ischemia. The ischemia-induced cellular K+ loss may be minimized by Kir4.1 downregulation and may contribute to the mechanism by which cellular acidification induces cell protection against ATP depletion. PMID- 17406123 TI - Novel PHEX mutation associated with hypophosphatemic rickets. AB - BACKGROUND: X-linked hypophosphatemia (XLH) is the most prevalent heritable form of rickets. It is a dominantly inherited disorder, characterized by renal phosphate wasting, abnormal vitamin D and PTH metabolism, and defective bone mineralization. Inactivating mutations in the gene encoding PHEX (phosphate regulating gene with homologies to endopeptidases on the X chromosome) have been found to be associated with XLH. METHODS: We report about a 54-year-old male patient who exhibited the typical features of XLH, and in whom mutational analysis using PCR and sequencing was performed. Additionally, extensive laboratory and radiological investigations were carried out. RESULTS: A 1-bp deletion in exon 2 of the PHEX gene was detected (177delC), which, to the best of our knowledge, has not been reported yet. This deletion results in a premature stop codon (C59X), suggesting a truncation of the PHEX protein. Furthermore, elevated FGF23 and PTH levels as well as an increased axial bone mineral density score were measured. CONCLUSIONS: We present a male patient with XLH, who harbors a novel mutation in the PHEX gene, which might be the cause for his disease. Our data support previous findings and therefore contribute to the decipherment of the pathogenetic pathways of XLH. PMID- 17406124 TI - P1,P4-diadenosine tetraphosphate (Ap4A) inhibits proximal tubular reabsorption of sodium in rats. AB - BACKGROUND/AIMS: P1,P4-diadenosine tetraphosphate (Ap4A) is a vasoactive dinucleotide possessing natriuretic activity. It is unclear, however, which part of the nephron is the target site of action for Ap4A. METHODS: We evaluated the tubular sites of Ap4A action using the lithium clearance technique. RESULTS: Ap4A at a priming dose of 2 micromol/kg with subsequent infusion at 20 nmol/kg/min increased fractional water and sodium excretion 2.5- and 5.6-fold, respectively. Moreover, Ap4A increased lithium clearance 1.9-fold and fractional lithium excretion 2.8-fold. Fractional water and sodium excretion from distal nephron segments was not significantly affected by Ap4A. CONCLUSION: These results suggest that Ap4A induces natriuresis mainly through inhibition of proximal tubular reabsorption of sodium. PMID- 17406125 TI - Adjuvant radiotherapy after radical prostatectomy: indications, results and side effects. AB - BACKGROUND: Within 5 years following radical prostatectomy, between 15 and 60% of patients with pT3 prostate carcinomas show an increasing prostate-specific antigen (PSA) level as a sign of local and/or systemic tumor progression. Apart from a large number of retrospective investigations, available results are present only from three randomized studies which have either been completely published or are only in abstract form. RESULTS: For pT3 prostate carcinomas the data from the three randomized studies agree, showing an around 20% reduced biochemical progression rate after 4-5 years. With these data the results of numerous retrospective studies have been confirmed. The majority of the authors use total doses of 60 Gy with single doses of 2 Gy. From one randomized study an increased local control rate is proposed as the basis for the extended freedom from biochemical progression. The rate of acute and late side effects after three dimensional radiotherapy with 60 Gy is very small and the rate of severe side effects is below 2%. The data for pT2 prostate carcinomas with positive margins are worse. Here controversy exists, and further investigations are required. In principle, however, adjuvant radiotherapy seems reasonable also for pT2 carcinomas with positive margins (determined by bNED - no biochemical evidence of disease). CONCLUSIONS: The effectiveness of adjuvant radiotherapy for patients with pT3 tumors and positive margins with and without detectable PSA levels is discussed. A survival advantage has not been demonstrated to date. For patients with positive margins in organ-limited prostate carcinomas (pT2 R1) randomized studies are recommended. It is unclear whether adjuvant radiotherapy is superior to radiotherapy for PSA levels increasing from the undetectable range after radical prostatectomy. To answer this question randomized studies are needed. PMID- 17406126 TI - Trans-obturator tape for incontinence: a 3-year follow-up. AB - AIM: To assess operative and functional outcomes, including morbidity, after using the trans-obturator tape for stress incontinence. PATIENTS AND METHODS: The first 24 consecutive patients undergoing trans-obturator tape insertion in a single centre were included in this retrospective study. All patients were female with a mean age of 63 (range 40-83) years. Fifteen patients (62.5%) suffered from pure stress incontinence, and 9 patients (37.5%) had mixed incontinence. Of the latter, 2 patients also had nocturnal enuresis. Each patient was followed up for between 3 and 12 months postoperatively and again at 36 months. The patients were assessed by clinical examination, ultrasound for residual urinary volume, and subjective satisfaction which was assessed at 3 and 12 months. At 36 months, all patients completed a validated incontinence questionnaire (International Consultation on Incontinence Questionnaire: ICIQ) which assessed female lower urinary tract symptoms and their impact on the quality of life. RESULTS: All operations were performed under epidural anaesthesia in day surgery, and the mean operative time was 20 (range 15-38) min. Nineteen patients (79.2%) showed significant improvement postoperatively, with 16 of those (66.6%) being completely cured of their incontinence. The remaining 5 patients (20.8%) were considered to have had failed procedures. There were no vascular, bladder, or urethral injuries. One patient had perforation of the vaginal fornix, and 3 patients developed vaginal erosion. CONCLUSIONS: The trans-obturator tape is a safe and effective treatment for stress incontinence and has a low morbidity after a 3-year follow-up period; however, it should only be performed by clinicians with the relevant surgical expertise and experience in treating female incontinence. PMID- 17406127 TI - Analysis of risk factors for urinary incontinence after radical prostatectomy. AB - INTRODUCTION: Our aim was to identify the independent risk factors associated with urinary incontinence after radical retropubic prostatectomy (RRP). MATERIALS AND METHODS: Using univariate and multivariate analyses, we examined several pre- and perioperative factors. One hundred and sixty-six patients were divided into three groups: patients who were immediately continent after catheter removal (group I), patients who became continent later (group II) and incontinent patients (group III). RESULTS: There were 34 patients (20.5%) in group I, 111 (66.9%) in group II, and 21 (12.6%) in group III. The multivariate analysis between the continent and incontinent patients proved that the postoperatively measured total length of the posterior urethra (strongly associated with length of the sphincter, length of the urethral stump and the presence of anastomotic stricture) was the independent risk factor for permanent incontinence or delayed continence following RRP. The age of patients per se represented a risk factor only for delayed continence, but not for permanent incontinence. CONCLUSIONS: Postoperatively measured shorter posterior urethral length results in an increased risk of urinary incontinence and delays continence after RRP. It seems that older age only delays reaching continence. PMID- 17406128 TI - Is there an impact of postoperative urethral and periurethral anatomical features in post-radical retropubic prostatectomy incontinence? AB - OBJECTIVES: To determine the anatomical changes in urethral and periurethral tissues after radical retropubic prostatectomy (RRP) by magnetic resonance imaging (MRI) and correlate these changes to continence. MATERIALS AND METHODS: 14 patients with grade 2-5 incontinence and 16 continent (grade 1) patients who had undergone non-nerve-sparing RRP for clinically localized prostatic carcinoma were evaluated. The sphincteric urethral length, transverse and craniocaudal diameters of both levator ani muscles and the volume of periurethral fibrosis were determined by abdominopelvic MRI. Continent (group 1) and incontinent (group 2) groups were compared in terms of sphincteric urethral length, transverse and craniocaudal diameters of levator ani muscles, volume of periurethral fibrosis, prostate volume, age and pathological stage. RESULTS: Among all the evaluated parameters, a statistically significant correlation was found only between the volume of periurethral fibrosis and the number of postoperative operations for urethral strictures in the incontinent group (p = 0.013, r = 0.859). CONCLUSIONS: Our results suggest that the postoperative sphincteric urethral length is not the sole factor in regaining continence after RRP as mentioned in the literature. PMID- 17406129 TI - Sixteen years of experience with stone management in horseshoe kidneys. AB - INTRODUCTION: Horseshoe kidney is the commonest congenital renal fusion anomaly, and is often complicated by urolithiasis. We focus on our 16 years of experience with stone management in horseshoe kidneys. MATERIALS AND METHODS: We reviewed the progress of 44 patients treated between 1987 and 2002. Shock wave lithotripsy (SWL) was used in 25 patients; the average stone surface area was 91 (range 10 1,600) mm2 and average follow-up was 36.5 (range 1-91) months. 19 patients underwent percutaneous nephrolithotomy (PCNL); the average stone surface area was 197 (range 6-2,400) mm2. Follow-up data are available for 8 patients and the average follow-up was 42.3 (range 3-144) months. RESULTS: In the SWL group the 3 month stone-free rate (SFR) was only 31%. In the PCNL group the SFR was 75% on the postoperative day-1 KUB. Complications occurred in 9 patients. CONCLUSIONS: Stone management in horseshoe kidneys is challenging: PCNL produces a higher SFR with minimal major complications and failed access. PCNL thus appears to be the preferred management option in patients with urolithiasis in horseshoe kidneys. PMID- 17406130 TI - Comparisons of urodynamic findings and voiding habits in patients with concomitant benign prostatic hyperplasia and detrusor overactivity presenting with or without the symptom of urgency. AB - AIMS: The urodynamic findings and voiding habits in patients with concomitant clinical benign prostatic hyperplasia (BPH) and detrusor overactivity (DO) presenting with or without the symptom of urgency were compared. MATERIALS AND METHODS: 84 BPH patients with an urodynamic diagnosis of DO by conventional cystometry were included in the study. The patients were grouped according to the presence or absence of the symptom of urgency. The urodynamic findings, urinary diary and clinical information were analyzed. RESULTS: Among the 84 BPH-DO patients, 52 reported the symptom of urgency while 32 did not. There were no significant differences in mean age, International Prostate Symptom Score and flow rate between the two groups. Patients without urgency had a higher incidence of terminal DO and abnormal bladder sensation. The occurrence of unfelt phasic DO was also significantly higher in this group. Sphincter electromyography showed conscious and subconscious sphincter contractions associated with DO. The urinary diary showed lower 24-hour urinary output, smaller bladder functional capacity and average voided volume in the BPH-DO patients without urgency. CONCLUSIONS: BPH patients with DO may neglect the symptom of urgency due to abnormal bladder sensation, or negate the symptom by subconscious sphincter contraction to abort the overactivity. Some may avoid the symptom by drinking less fluid and emptying the bladder at a smaller volume. PMID- 17406131 TI - Is ultrasound of the retroperitoneum a valuable staging method in selecting testicular cancer patients for primary retroperitoneal lymph node dissection? AB - OBJECTIVE: In nonseminomatous testicular cancer patients with normal serum tumor markers and no distant metastasis, postorchiectomy surgery is a valid treatment option if the disease extension into the retroperitoneum is not advanced. We assessed the ability of ultrasound (US) to exclude the presence of bulky retroperitoneal disease. MATERIALS AND METHODS: One hundred and forty testicular cancer patients underwent US and computed tomography (CT) of the retroperitoneum. US results were analyzed using three cutoffs: 5 cm (conventional staging), 3 cm (based on the minimal sonographical dimension of actual bulky disease) and 0 cm ('clean retroperitoneum' or any detectable nodes), and were compared with CT data using the 5-cm cutoff ('gold standard'). RESULTS: The sensitivity, specificity, overall accuracy, positive and negative predictive values of US in detecting of bulky retroperitoneum for the 5-cm cutoff were 83, 96, 93, 88 and 94%, for the 3 cm cutoff 100, 91, 94, 80 and 100%, and for the 0-cm cutoff 100, 66, 74, 49 and 100%, respectively. CONCLUSIONS: In stage I and IIA/B marker-negative nonseminomas if the treatment strategy is surgery, US may facilitate the selection process; the report of a clean retroperitoneum safely excludes the presence of bulky disease and may be an indication for lymphadenectomy, although in case of positive findings a CT should be performed. PMID- 17406132 TI - Citrate in expressed prostatic secretions has the feasibility to be used as a useful indicator for the diagnosis of category IIIB prostatitis. AB - INTRODUCTION: Normal human prostatic secretion is remarkably rich in citrate (EPS citrate). Prostate inflammation will result in reduction of EPS-citrate level. So EPS-citrate was determined to investigate whether EPS-citrate level was also decreased in category IIIB prostatitis and whether it is feasible for EPS-citrate to be used as a diagnostic marker for category IIIB prostatitis. MATERIALS AND METHODS: 21 category II outpatients, 25 category IIIA outpatients, 25 category IIIB outpatients and 21 normal controls without any prostatic diseases were included. All category II, IIIA and IIIB outpatients had not received any management. The EPS-citrate was determined by high-performance liquid chromatography. RESULTS: The mean EPS-citrate values were 3.32 +/- 0.79, 3.41 +/- 0.88, 4.37 +/- 0.77 and 8.55 +/- 1.20 mg/ml in groups II, IIIA, IIIB and normal controls, respectively. Compared to normal controls, the EPS-citrate levels were significantly decreased in category II, IIIA and IIIB patients. Furthermore, there was no overlap of EPS-citrate values between category IIIA and IIIB patients and the normal control group. CONCLUSIONS: The EPS-citrate level is decreased in category IIIB prostatitis and it is feasible for EPS-citrate to be used as a useful diagnostic indicator for category IIIB prostatitis. PMID- 17406133 TI - Adult male circumcision outcomes: experience in a developing country setting. AB - INTRODUCTION: We examined male circumcision outcomes among young adults in an African setting. MATERIALS AND METHODS: Participants were healthy, sexually active, uncircumcised, HIV-seronegative males aged 18-24 years. The main outcomes measured included complications, healing, satisfaction and resumption of activities. RESULTS: Of 1,475 procedures, 26 (1.8%) were associated with 27 adverse events, most commonly wound disruption/delayed healing (0.6%), wound infection (0.4%), and bleeding (0.3%). Adverse events per clinician averaged 3.8 and 2.1% for procedures 1-100 and 101-200, respectively, and <1% for procedures 201-300, 301-400 and >400, respectively (p < 0.001). Participants resumed normal general activities after a median of 1 postoperative day and 93% with regular employment resumed working within 1 week. After 30 days, 99% of participants reported being very satisfied. After 90 days, 65% reported having had sex, 45% reported that their partners had expressed an opinion, 92% of whom were very satisfied with the outcome. CONCLUSIONS: Safe and acceptable adult male circumcision services can be delivered in developing country settings. PMID- 17406134 TI - Dorsal double-layer dartos flap for preventing fistulae formation in the Snodgrass technique. AB - INTRODUCTION: The Snodgrass technique and its modifications have become a preferred method for all varieties of hypospadias in the past decade. However, fistula is the most common complication of this technique. The aim of this study was to investigate the importance of the single and double flap to prevent fistula formation in the Snodgrass procedure. MATERIALS AND METHODS: Tubularized incised plate urethroplasty, using a single or the double flap, was undertaken in 74 consecutive boys (median age 6.6 years old, range 1-15) within the last 4 years. In the first 29 patients (group 1), a dorsolateral flap was rotated laterally for covering the neourethra and in the remaining 45 patients (group 2) the neourethra was covered with dorsal double dartos flaps. RESULT: In group 1, fistula in 4 patients and partial glanular dehiscence in 1 patient were detected. There was no fistula formation in group 2. CONCLUSION: For preventing fistula formation, urethral covering by a well-vascularized dorsal double-layer dartos flap should be the basic part of the Snodgrass procedure. PMID- 17406135 TI - Value of urinary flow patterns in the follow-up of children who underwent Snodgrass operation. AB - The urinary flow patterns and the Toguri nomogram were compared in the intermediate functional results of the tubularized-incised plate urethroplasty (TIPU) to repair distal and midpenile hypospadias by using uroflowmetry. 28 children who were toilet trained, were able to void volitionally, and had no fistulas following hypospadias repair were eligible for the study. The study did not include children who had persistent fistula, meatal stenosis or urethral stricture, and did not return for follow-up. The mean age was 8.4 years and the mean follow-up period was 18 months. The urinary flow pattern, maximum (Q(max)) and average flow rate (Q(ave)) were measured; the results were expressed as percentiles and compared to the Toguri values from normal children. The Q(max) and Q(ave) were considered normal if they were in >25th percentile, equivocally obstructed in the 5-25th percentile and obstructed if <5th percentile. The flow pattern was classified as bell ring shape, plateau or intermittent. According to the Toguri nomogram, 22 of 28 patients (78.5%) were considered normal, 4 patients (14.2%) as equivocally obstructed, and 2 patients (7.1%) as obstructed group. A normal bell-shaped flow curve was obtained in 23 (82.1%) of the children. 4 patients (14.2%) had a plateau flow pattern. Only 1 of the patients had an intermittent shape flow curve. The flow pattern was normal bell-shaped for all of the patients, except 1, with Q(max) above the 25th percentile according to the Toguri nomogram. Of children with Q(max) below the 5th percentile, both of them had a plateau flow pattern and were found to have an asymptomatic meatal stenosis, which was improved with urethral dilatation. However, of the 4 patients with Q(max) between 5 and 25 percentiles, 2 had a plateau flow pattern and the others had a bell-shaped flow pattern. The flow patterns of the 2 patients determined as obstructive by the Toguri nomogram were plateau-shaped. TIPU provides satisfactory functional results for distal and midpenile hypospadias; uroflowmetry is an important noninvasive tool to evaluate this technique. There are no studies in the literature which only used flow patterns for the evaluation of urination for follow-up after the hypospadias repair. Our study showed that the evaluation of obstruction according to the Toguri nomogram may not be necessary in patients with a normal bell-shaped flow pattern in uroflowmetry. PMID- 17406136 TI - Healing of the urethral plate after deep incision: does catheterization change the course of this process? AB - BACKGROUND: We aimed to evaluate the sequence of healing process as well as possible effects of stent placement on the healing process after deep urethral plate incision. METHODS: A deep urethral plate incision was done at the 12 o'clock position. After that, in the first group (n = 14) the anterior urethra was stented with a silicon catheter. Animals in the second group (n = 14) underwent the same incision procedure, however no stent was placed after this operation. All animals in both groups were again divided into three groups with respect to the follow-up period (7-14 and 21 days). Partial penectomy was performed in all subgroups and histopathologic evaluation performed. RESULTS: In the first group after 7 days, limited neovascularization and granulation tissue formation could be noted far away from the epithelial lumen. Evaluation of these specimens during the long-term follow-up (21 days) demonstrated an almost completely healed tissue with a remarkable neovascularization and well-developed granulation tissue. In the second group during 14-21 days, evaluation progression of tissue healing along with increasing vessel formation and re-epithelialization were demonstrated. Although the incision edges did show evident approximation, no sign of fibrosis could be demonstrated in these specimens. CONCLUSION: We may say that tissue healing with a desired and complete re-epithelialization could be achieved without inserting a catheter. Prevention of re-approximation along with the limited urinary extravasations to the subepithelial area might be responsible for tissue protection that will limit the long-term aforementioned adverse effects of the procedure. PMID- 17406137 TI - A new antireflux technique: mucosal flap barrier. AB - INTRODUCTION: In this study, our aim was to evaluate the effectiveness of the mucosal flap barrier (MFB) upon the incompetent ureteral orifice of an experimentally produced rabbit reflux model. The rabbit reflux model was done in the first stage of the study. This method is a new and original procedure which is different from Bischoff's and Witherington's osteoplasty techniques. MATERIALS AND METHODS: We performed the mucosal flap procedure in eight 2-2.5 kg male New Zealand rabbits with experimentally produced refluxing ureters. The first stage of our study was to create refluxing ureters. Due to the upward and inward 'J' fishhook course of the intravesical ureter of rabbits, we performed two deep 12- and 4-o'clock incisions by suturing the edges of the intravesical ureter apart with catgut sutures. Additionally, to stop the tilting action of the trigonum during micturition, we performed two parallel incisions between the ureteral meatus. These parallel incisions were the authors' modifications for the rabbit reflux model and which gives long-lasting reflux for the rabbit. We checked the reflux at the end of the 3rd month and performed our new MFB antireflux procedure. This mucosal flap made a cover on the incompetent ureteral orifice. This technique is completely different from the osteoplasty techniques described by Bischoff and Witherington. RESULTS: Four months after the MFB application, we performed intravenous urography, cystometry and cinefluorography. There was no obstruction or reflux. Two rabbits were sacrificed at the 4th month. Both flaps were fully intact without any fixation at the medial side causing obstruction. At present, 3 rabbits are still alive. We did cinefluorography and cystometry at the 15th month and there was no obstruction or reflux. We are trying to keep them alive till the end of their natural lifespan. CONCLUSIONS: During the formation of the rabbit reflux model, we observed that spontaneous reflux even at high intravesical pressures did not have high percentages as previously mentioned in the literature. But it is possible and enough to produce an experimental reflux model by using meatotomy and interureteric incisions. Moreover, in rabbits with vesicoureteral regurgitation produced by ureteral meatotomy and interureteral incisions, reflux can be eliminated the MFB procedure. This method is an easy procedure for small bladders or dilated ureters after any kind of open reimplantation applications and it could be used by pneumovesical applications in the future. PMID- 17406138 TI - Is second-line enuretic alarm therapy after unsuccessful pharmacotherapy superior to first-line therapy in the treatment of monosymptomatic nocturnal enuresis? AB - INTRODUCTION: We aimed at comparing the success rates of primary enuretic alarm therapy with those of secondary alarm therapy after failed pharmacotherapy in the treatment of monosymptomatic nocturnal enuresis (MNE). PATIENTS AND METHODS: We randomly applied enuretic alarm therapy in 35 MNE patients (group 1) and desmopressin therapy in 49 MNE patients (group 2). The success and rebound rates after 3 and 6 months were determined. We also applied enuretic alarm therapy as a secondary treatment in 19 group 2 patients with complete rebound after 6 months (group 3). The success rates of patients who have received primary and secondary enuretic alarm therapy were compared. RESULTS: The success rates for groups 1 and 2 were 82.65 and 81.63%, respectively (p = 0.885), at 3 months and 54.28 and 26.53%, respectively (p = 0.007), at 6 months. The success rates in group 3 were 84.21 and 52.63%, respectively, at 3 and 6 months. When these success rates were compared between groups 1 and 3, no statistically significant difference was found (p = 1.000). CONCLUSION: Prior pharmacotherapy did not increase success rates of alarm therapy in our MNE patients. PMID- 17406139 TI - A review of the natural progression of medullary sponge kidney and a novel grading system based on intravenous urography findings. AB - PURPOSE: Medullary sponge kidney (MSK) is a developmental abnormality of the medullary pyramids of the kidney, characterised by cystic dilatations of the collecting ducts. We investigated a cohort of patients with MSK to gain further information about its presentation, clinical course and treatment required. We devised a grading system based upon findings at diagnostic intravenous urography (IVU) and established a relationship between severity of IVU findings and severity of disease. MATERIALS AND METHODS: The clinical notes and imaging of 29 patients with MSK were analysed. The severity of IVU findings was classified as follows: grade 1 (one calyx, unilateral), grade 2 (one calyx, bilateral), grade 3 (more than one calyx, unilateral) and grade 4 (more than one calyx, bilateral). RESULTS: The age range at diagnosis was 12-69 (mean 39), mean follow-up period was 12.7 years. Increasing grade of IVU findings correlated with more frequent symptomatic stone episodes (grade 1: 0.09 episodes per patient per year; grade 4: 0.34). Higher grade was also related to the number of hospital admissions (grade 1: 0.182 per patient per year; grade 4: 0.282) and the number of procedures required (either surgery or extracorporeal shock wave lithotripsy; grade 1: 0.0 interventions per patient per year; grade 4: 0.24). CONCLUSIONS: MSK presents over a wide age range and can cause long periods of intermittent episodes. Patients can be graded using a novel system based on IVU findings, which correlates with severity of disease. Management strategies such as intensity of follow-up can be modified using this grading. PMID- 17406140 TI - Upregulation of heme oxygenase and collagen type III in the rat bladder after partial bladder outlet obstruction. AB - The objective of the study was to evaluate possible changes of the gene expression and localization of the enzymes, heme oxygenase and nitric oxide synthase (NOS), with reference to increase of collagen type III in response to the partial obstruction of the bladder. Following initial obstruction, whole rat bladders were removed for real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Real-time RT-PCR demonstrated significantly enhanced expression of HO (p < 0.01) and collagen type III (p < 0.001) gene on postoperative day 14. Enhanced expression of NOS gene was seen only on postoperative day 4 (p < 0.01). Immunohistochemistry revealed that immunoreactivity to HO-1 had much in common in neural cells and fibers, although immunoreactivity to HO-2 and iNOS was relatively weak. This study suggested gene expression of HO, especially HO-1, was more dramatically changed than NOS, and was upregulated simultaneously with increase of collagen type III after obstruction. HO systems could be involved in the pathogenesis of bladder dysfunction related to increase of collagen type III after obstruction. PMID- 17406141 TI - Juvenile granulosa cell tumor of the epididymis. AB - We report the first case of a juvenile granulosa cell tumor of the epididymis in a young man. Clinical and histological findings are presented and the clinical significance of the case is discussed. PMID- 17406142 TI - Cutaneonephrobronchial fistula: an unusual presentation of an inadequately treated perinephric abscess. AB - We report a case of cutaneonephrobronchial fistula secondary to an inadequately treated perinephric abscess associated with a staghorn calculus. The patient, a 72-year-old lady, was initially treated for a superficial flank abscess by incision and drainage. She returned 4 months later with a persistent discharging flank sinus, at which time the above complex pathology was diagnosed. We review similar cases in the literature and discuss pathophysiology and current management of perinephric abscesses. PMID- 17406143 TI - De novo renal cell carcinoma in a kidney allograft 13 years after transplantation: a case report and review of the literature. AB - De novo carcinoma of the renal transplant is a rare but disastrous clinical entity. We report such a tumor developing 13 years after transplantation and describe its clinical presentation, diagnostic approach and therapy. The importance of a surveillance program allowing early detection of tumor developing in the renal transplant is emphasized. PMID- 17406144 TI - Metastatic primitive neuroectodermal tumor to the kidney. AB - Primitive neuroectodermal tumors of the kidney are rare neoplasms that may mimic renal cell carcinoma, especially in the presence of locally advanced or metastatic disease. Although several cases have been identified as primary renal tumors, this is the first report of metastatic spread of a primitive neuroectodermal tumor to the kidney. PMID- 17406145 TI - Primary alveolar soft part sarcoma of chest wall: a case report and review of the literature. PMID- 17406146 TI - Multivitamin use in relation to self-reported body mass index and weight loss attempts. AB - CONTEXT: There are scant data on patterns of multivitamin use among US adults in terms of body mass index (BMI) or whether one is trying to lose weight. OBJECTIVE: To examine multivitamin use and beliefs about multivitamin use among adults according to BMI and to determine whether use by body weight differs if one is trying to lose weight. DESIGN: Cross-sectional multivariate analysis of the HealthStyles consumer survey. The final analytic sample consisted of 2239 women and 1532 men. MAIN OUTCOME MEASURES: Prevalence and odds of multivitamin use by demographic and behavioral characteristics including BMI, use by weight loss intent, and among users, reasons for use. RESULTS: 63.7% of women and 52.9% of men reported multivitamin use (taking 1 or more multivitamin per week). Obese women were less likely than normal-weight women to use multivitamins; no differences according to BMI category were detected for men. Among women who were not trying to lose weight, obese women were less likely than normal-weight women to use multivitamins (odds ratio = 0.63, CI 0.41-0.98). Assessment of reasons for use found that compared among women not trying to lose weight, those trying to lose weight were more likely to report multivitamin use because It is important for my health. CONCLUSIONS: This descriptive analysis adds to the limited literature on multivitamin use according to both body weight and attempting to lose weight. Multivitamin use was common and decreased with increasing BMI. This may be because fewer obese people consider vitamins "important for their health". PMID- 17406147 TI - The heart and soul of the American Medical Association. PMID- 17406148 TI - Ewing's sarcoma metastasis to the gastric wall in a 72-year-old patient. PMID- 17406149 TI - Thromboendarterectomy as treatment in the antiphospholipid syndrome. AB - Antiphospholipid antibody syndrome (APS) may lead to pulmonary hypertension, a lethal complication of chronic pulmonary thromboembolism. There are few viable treatment options for pulmonary hypertension, but pulmonary thromboendarterectomy (PTE) has been used to successfully treat this condition in many patient populations. Following is a description of a patient with APS and systemic lupus erythematosus (SLE) who presented with severe pulmonary hypertension and underwent lifesaving surgery with pulmonary thromboendarterectomy. Successful intervention with thromboendarterectomy for the treatment of severe pulmonary hypertension is possible in patients with the antiphospholipid antibody syndrome, and surgical referral of these patients is warranted. PMID- 17406150 TI - Hypocitraturia despite potassium citrate tablet supplementation. AB - Citrate supplementation is widely used in the prevention of recurrent nephrolithiasis with hypocitraturia. Potassium citrate is the most commonly used citrate agent for this indication. In patients with chronic diarrheal syndromes, the absorption of potassium citrate can be affected. We describe a patient who presented with recurrent nephrolithiasis and chronic diarrhea and was found to have severe hypocitraturia despite citrate supplementation with potassium citrate tablets, likely due to inadequate gastrointestinal absorption of citrate from the slow-release wax-matrix tablets. PMID- 17406151 TI - The high cost of money-driven medicine. PMID- 17406152 TI - A decade (or so) of the medical Internet: are we getting anywhere? PMID- 17406153 TI - Massive primary hepatic tuberculoma mimicking hepatocellular carcinoma in an immunocompetent host. AB - Tuberculoma of the liver is rare in an immunocompetent individual. We report a 26 year-old man with upper abdominal pain, abnormal liver function, and raised inflammatory markers. Abdominal computed tomography (CT) scan revealed a mixed attenuation lesion measuring 6 x 5 cm occupying most of the left lobe of the liver. Subsequent histology and culture confirmed tuberculous abscess. Following antituberculous therapy, repeat CT scan revealed complete resolution of the initial findings. This case illustrates the diagnostic difficulties of hepatic tuberculosis (TB) and the importance of considering TB in patients with hepatic lesions. PMID- 17406154 TI - The effects of attention-deficit/hyperactivity disorder on employment and household income. AB - CONTEXT: Many children with attention-deficit/hyperactivity disorder (ADHD) continue to exhibit symptoms of the disorder into adolescence and adulthood. Although ADHD may have a profound impact on activities of daily living, including educational achievement and work performance, limited research exists on ADHD's impact on individual income loss and overall economic effect. OBJECTIVES: Evaluate ADHD's impact on individual employment and income, and quantify costs of ADHD on workforce productivity for the US population. DESIGN: Two telephone surveys were conducted between April 18, 2003, and May 11, 2003, to collect demographic, educational, employment, and income information. PARTICIPANTS: Two groups of adults aged 18-64 years were interviewed: those diagnosed with ADHD (n = 500) derived from a national list of mail-paneled members who identified themselves or a household member as having been diagnosed with ADHD, and an age- and gender-matched control group (n = 501) derived from a random digital-dialing sample of a national cross-section not diagnosed with ADHD. RESULTS: Statistically fewer subjects in the ADHD group achieved academic milestones beyond some high school (P < .05). In addition, fewer subjects with ADHD were employed full time (34%) compared with controls (59%; P < .001). Except for the subgroup of subjects aged 18-24 years, average household incomes were significantly lower among individuals with ADHD compared with controls, regardless of academic achievement or personal characteristics. On the basis of these findings, loss of workforce productivity associated with ADHD was estimated between $67 billion and $116 billion. CONCLUSIONS: Decreased individual income among adults with ADHD contributes to substantial loss in US workforce productivity. PMID- 17406156 TI - Is national health insurance coming to the United States? PMID- 17406157 TI - The medical home: its time has come! PMID- 17406158 TI - Differences between the perceived and actual age of overweight onset in children and adolescents. AB - OBJECTIVE: Little is known about whether children or their parents can accurately recall the age at which they became overweight. DESIGN, SUBJECTS AND MAIN OUTCOME MEASURES: We interviewed 64 overweight children (7-18 years old) about their weight history and compared reported age of overweight onset with actual onset, as determined by the age at which the child's measured BMI first exceeded the 95th percentile. RESULTS: Only 28% of children reported overweight onset within 1 year of actual overweight onset. Reported overweight onset age (7.6 +/- 2.5 y) and actual onset age (5.3 +/- 2.5 y; P < .001) were not significantly correlated (r2 = .03, P = .22). Children who became overweight before 8 years of age tended to report becoming overweight at a later age than actual onset, whereas children who became overweight after 8 years of age tended to report becoming overweight at an earlier age than actual onset (P < .001), with 27% of children either underreporting or overreporting their overweight onset by at least 5 years. Similar results were found when analyzing parent reports of their children's overweight onset. CONCLUSIONS: Reported and actual overweight onset ages were uncorrelated in our sample, suggesting that families may not be cognizant of children's growth trajectories. Greater efforts should be made to help parents and children understand and track growth patterns with the aim of preventing excessive weight gain. PMID- 17406159 TI - Case 13: a man with progressive headache and confusion. PMID- 17406161 TI - Should decisions about your hiring, reappointment, tenure, or promotion use the impact factor score as a proxy indicator of the impact of your scholarship? PMID- 17406160 TI - IBS--review and what's new. AB - Irritable bowel syndrome (IBS) is a highly prevalent gastrointestinal motility disorder broadly characterized by abdominal pain/discomfort associated with altered bowel habits. The chronic and bothersome nature of IBS symptoms often negatively affects patient quality of life and activity level and places a substantial economic burden on patients and the healthcare system. Advances in research have led to a greater understanding of the underlying pathophysiology of IBS, particularly regarding the role serotonin plays in the gastrointestinal tract; the development of stepwise, symptom-based diagnostic strategies that allow for a diagnosis of IBS to be made without the need for extensive laboratory testing; and the development of treatment options targeting underlying pathophysiologic mechanisms that provide relief of the multiple symptoms associated with IBS. This review highlights recent advances in research and discusses how these findings can be applied to daily clinical practice. PMID- 17406162 TI - A new aggressive approach to screening and early intervention to prevent death from coronary artery disease. PMID- 17406163 TI - Device-guided breathing to lower blood pressure: case report and clinical overview. AB - The belief that breathing exercises may provide health benefits has been shared by many cultures for centuries. A case study illustrates one such FDA-approved intervention, its performance over time, and the day-to-day home blood pressure (BP) variation in response to treatment. The device used by this patient (which interactively entrains slowed and deep breathing) has been studied in 7 clinical trials. Routine use of the device (RESPeRATE, InterCure Inc., Fort Lee, New Jersey; www.resperate.com/MD) significantly lowered home and office BPs without adverse effects, when used alone, with lifestyle modifications, or adjunctively to antihypertensive drugs. The proposed physiological mechanism(s), the technology that guides slowed breathing, pooled clinical research outcomes, and recommendations regarding this modality in clinical practice are also reviewed. Device-guided paced breathing may offer an effective, simple, and new nonpharmacologic option for treating high BP without additional side effects, but like all lifestyle modifications, must be practiced consistently to provide benefits. PMID- 17406164 TI - The hospitalist movement 10 years later: life as a Swiss army knife. PMID- 17406165 TI - The reality of "traditional Chinese" medicines. PMID- 17406166 TI - Case files from Stanford University Medical Center: Drug resistance testing in previously untreated patients with HIV--knowing what to look for and choosing appropriate therapy. PMID- 17406167 TI - A comparison of the lidocaine patch 5% vs naproxen 500 mg twice daily for the relief of pain associated with carpal tunnel syndrome: a 6-week, randomized, parallel-group study. AB - OBJECTIVES: Carpal tunnel syndrome (CTS) is a common entrapment neuropathy caused by median nerve compression. This pilot clinical trial was designed to compare the safety and effectiveness of the lidocaine patch 5% to that of naproxen 500 mg twice daily for the treatment of neuropathic pain associated with CTS. METHODS: In this 6-week, randomized, parallel-group, open-label, multicenter study, participants from 2 practice sites, aged 18 to 75 years with clinical/electrodiagnostic evidence of CTS, were randomized to receive up to 3 lidocaine 5% patches every 24 hours or naproxen 500 mg twice daily for 6 weeks. Outcome assessments included mean changes between baseline and Week 6 average pain intensity (Brief Pain Inventory [BPI]: Question 5, Average Pain Intensity [API]), an Investigator Clinical Global Impression of Improvement (CGI-I) over the course of the treatment period and a comparison of patient satisfaction (Clinical Global Assessment of Treatment [CGAT]). RESULTS: One hundred patients were randomized in this study, 52 in the lidocaine patch 5% group and 48 in the naproxen 500 mg twice daily group. Significant reductions in API scores were observed between baseline and Week 6 for both lidocaine patch 5% (P < .0001) and naproxen 500 mg twice daily (P = .0004); however, there were no statistically significant differences between treatments (P = .083). There was a significant (P = .016) difference in the CGI-I for lidocaine patch 5% (51.1%) compared with naproxen 500 mg twice daily (24.3%). Whereas 71.8% of the lidocaine patch 5% patients reported being "satisfied" to "very satisfied" with the treatment, only 63.2% of naproxen 500 mg twice daily patients reported likewise, although the difference was not statistically significant. Both treatments were well tolerated. Two patients reported treatment-related adverse events in the lidocaine patch 5% group and 6 in the naproxen 500 mg twice daily group, all of which were considered mild or moderate in severity. CONCLUSIONS: This study demonstrates that the lidocaine patch 5% is effective in significantly relieving the pain associated with CTS and is well tolerated. The patch may offer patients an effective, nonsystemic, noninvasive treatment for the management of their symptoms. Further controlled studies are warranted. PMID- 17406168 TI - Multiple penile schwannomas. AB - The occurrence of penile schwannoma is very rare. A 41-year-old man presented with multiple penile tumors and pain on erection. The largest tumor causing pain was excised. Pathology was characteristic of benign schwannoma. We recommend that penile schwannomas be excised if the tumors cause pain or are malignant. PMID- 17406169 TI - The future of emergency care in the United States. PMID- 17406170 TI - Time to reform physician relicensure. PMID- 17406171 TI - Predictors of bipolar disorder risk among patients currently treated for major depression. AB - OBJECTIVE: The primary objective of this study was to assess the rate of bipolar disorder (BPD) risk among patients unsuccessfully treated for major depression and to identify predictors of bipolarity. PATIENTS, DESIGN, AND SETTING: Psychiatrists from community and private practice settings sequentially selected patients with unipolar depression who exhibited nonresponse to at least 1 antidepressant (AD) trial. MAIN OUTCOME MEASURES: Patients self-reported their demographics, family history, comorbid health status, and legal problems. Current depression symptoms were assessed via the Centers for Epidemiologic Studies- Depression (CES-D) scale. Bipolar screening was performed using the Mood Disorder Questionnaire (MDQ). A psychiatrist recorded patient history and current and prior AD medication use. RESULTS: Of 602 patients enrolled, 18.6% screened positive on the MDQ. This rate was not affected by the number of prior AD failures or patient demographics. A prior history of BPD was reported by 12.3% of patients of which the psychiatrist was not aware or did not report, and 41.2% of these patients were MDQ+. Stepwise logistic regression identified 5 variables associated with bipolar disorder risk: the CES-D item "people were unfriendly," comorbid anxiety, initial depression diagnosis within 5 years, family history of BPD, and legal problems. In the subset of patients with complete data for the 5 variables (n = 483), 41.3% of the patients endorsing any 3 or more risk factors (n = 109) were MDQ+. CONCLUSIONS: Almost 20% of patients with AD nonresponsive unipolar depression screened positive for BPD and the number of past medication trials had no effect on bipolarity. This suggests that clinicians should carefully screen for BPD in patients who have failed at least 1 antidepressant. Comorbid anxiety, feelings of people being unfriendly, recent depression diagnosis, BPD family history, and legal problems may prove useful indicators of BPD risk among patients who have failed at least 1 antidepressant. Future studies are needed to confirm that these risk factors are useful clinical variables to screen for bipolarity. PMID- 17406172 TI - Diet variety based on macronutrient intake and its relationship with body mass index. AB - CONTEXT: Increased dietary variety has been associated with increased body weight; however, diet variety is not measured using a standardized methodology. OBJECTIVE: We developed a new dietary variety score (DVS) based on food macronutrient content, and determined the relationship between DVS and measures of adiposity. DESIGN: Cross-sectional, observation study. SETTING: University of Alabama at Birmingham (UAB), EatRight Weight Management Program. PATIENTS: Study subjects (n = 74) were former participants of UAB's EatRight Weight Management Program who completed the program at least 1 year ago. MAIN OUTCOME MEASURES: Unique items from 4-day food records were converted to macronutrient categories using the diabetic exchange. Foods were categorized as a source of carbohydrate if containing > or = 5 g carbohydrate and > or = 20 calories; fat if containing > or = 5 g fat; protein if containing > or = 7 g protein. Height and weight were measured; BMI (kg/m2) was calculated. Dual energy X-ray absorptiometry measured body fat. Generalized linear modeling in SAS (Ver.9) determined relationships between adiposity and DVS. RESULTS: On average, participants weighed 92.7 -/+ 30.9 kg (BMI = 32.5 kg/m2). Men and women's body fat were 37.4 -/+ 6.4% and 47.3 /+ 6.4%, respectively. Separate linear regression models containing terms for sex and DVS showed significant relationships between macronutrient DVS and BMI. In a multivariate model controlling for each of the macronutrient DVS, BMI was not related to fat DVS, but was found to be positively related to protein DVS (beta = 0.87, P = .04). Sex modified the relationship between carbohydrate DVS and BMI, with an inverse relationship between BMI and carbohydrate DVS among women, and a positive relationship among men. Percent body and trunk fat were not related to DVS for either gender. CONCLUSIONS: The new DVS, based on macronutrient content of foods, had significant associations with BMI. These dietary variety scores may provide another way to evaluate the impact of consuming a variety of food types on energy intake and BMI. PMID- 17406173 TI - What are patients looking for? PMID- 17406174 TI - Grand rounds in gastroenterology from Baylor College of Medicine. Upper gastrointestinal bleeding in the ICU. PMID- 17406175 TI - Boosting the odds of surviving cardiac arrest. PMID- 17406178 TI - We need to do a better job of preparing for the next big disaster. PMID- 17406177 TI - Rethinking global health training in North America. PMID- 17406176 TI - Comparative neurocognitive effects of 5 psychotropic anticonvulsants and lithium. AB - CONTEXT: Many of the new antiepileptic drugs have psychiatric indications, and most are prescribed by psychiatrists for patients with mood disorders, even when a specific indication is absent. Epileptic drugs as a whole, even the newer ones, are known to affect cognition, sometimes in untoward ways. Research on the neurocognitive effects of antiepileptic drugs, however, has been done exclusively in normal volunteers and in patients with seizure disorders. METHOD: A naturalistic, cross-sectional study was conducted on patients who were taking 1 of 5 different antiepileptic drugs or lithium (LIT). Cognitive status was measured by a computerized neurocognitive screening battery, CNS Vital Signs (CNSVS). SUBJECTS: One hundred fifty-nine patients with bipolar disorder, aged 18 70 years, were treated with carbamazepine (CBZ) (N = 16), lamotrigine (LMTG) (N = 38), oxcarbazepine (OCBZ) (N = 19), topiramate (TPM) (N = 19), and valproic acid (VPA) (N = 37); 30 bipolar patients were treated with LIT. RESULTS: Significant group differences were detected in tests of memory, psychomotor speed, processing speed, reaction time, cognitive flexibility, and attention. Rank-order analysis indicated superiority for LMTG (1.8) followed by OCBZ (2.1), LIT(3.3), TPM (4.3), VPA (4.5), and CBZ (5.0). CONCLUSION: The relative neurocognitive effects of the various psychotropic antiepileptic drugs in patients with bipolar disorder were concordant with those described in the seminal literature in normal volunteers and patients with epilepsy. LMTG and OCBZ had the least neurotoxicity, and TPM, VPA, and CBZ had the most. LIT effects on neurocognition were intermediate. Choosing a mood-stabilizing drug with minimal neurocognitive effects may enhance patient compliance over the long term. PMID- 17406179 TI - Quality consortia: what they do. PMID- 17406180 TI - Institute for quality in laboratory medicine series--controversies in laboratory medicine: nursing and the laboratory: relationship issues that affect quality care. PMID- 17406182 TI - The epidemic of chronic kidney disease. PMID- 17406181 TI - Tackling COPD: a multicomponent disease driven by inflammation. AB - In recent years, research has revealed more about the factors underlying the pathogenesis of chronic obstructive pulmonary disease (COPD). In particular, inflammation in the lungs leads to the structural changes observed in COPD, while extrapulmonary symptoms and comorbidities may be systemic manifestations of these inflammatory processes. A new multicomponent disease model is proposed that takes into account all elements that should be considered in treatment decisions. Current monotherapies act on different aspects of COPD and may not address all components. A combination of a long-acting beta2 agonist and an inhaled corticosteroid has complementary effects, addressing a wider range of components of COPD. This combination appears to have greater clinical benefits than either agent alone in reducing the frequency of exacerbations, reducing the number of hospitalizations, and potentially promoting survival. Minimizing the burden of COPD within--and potentially outside--the lung means treating patients early and addressing as many disease components as possible. PMID- 17406183 TI - Evaluation and treatment of hepatitis C in patients with coexisting HIV infection. PMID- 17406184 TI - Hidrocystomas--a brief review. AB - Hidrocystomas, eccrine and apocrine, are rare cystic lesions that form benign tumors of the sweat glands. In this clinical review, we present a "classic" case of hidrocystoma and review the main epidemiologic, histologic, and clinical features. We also review the effectiveness of experimental treatment methods and present information about associated syndromes and differential diagnosis, focusing especially on hidrocystomas' resemblance to basal cell carcinoma. PMID- 17406185 TI - Trends in childhood anemia in a Massachusetts Health Maintenance Organization, 1987-2001. AB - CONTEXT: An important indicator of nutritional status within a pediatric population is the anemia prevalence rate. Limited national data are available regarding trends in anemia prevalence among non-low-income children. OBJECTIVE: To determine the prevalence of anemia over time among children enrolled in a health maintenance organization (HMO). DESIGN: Trend analysis, adjusted for clinical site, age group, sex, race/ethnicity, Medicaid insurance, and testing frequency within each year, and accounting for repeated observations of individual children across different years. SETTING: Massachusetts HMO. PARTICIPANTS: 72,729 children aged 6-59.9 months seen at 126,695 well-child visits from 1987 to 2001. MAIN OUTCOME: Anemia prevalence by hemoglobin level. RESULTS: The observed prevalence of anemia was 8.9%, and was higher among younger children, blacks, boys, and those with Medicaid insurance. While the unadjusted anemia rate among children tested increased from 9.9% in 1987 to 11% in 2001, the proportion of children who were tested for anemia declined from 55% to 45% during the study period. After adjustment for demographics and testing frequency, the odds of anemia decreased over time (odds ratio 0.84, 95% confidence interval 0.76 0.93, per decade). Predicted anemia prevalence among those tested decreased from 8.9% in 1987 to 7.1% in 2001. CONCLUSIONS: Anemia was less prevalent in this HMO population than has been reported in low-income children. After adjustment for testing frequency, the odds of anemia declined over time. As testing practices change to target children at higher risk, trend analyses should account for differences in screening rates over time. The computerized medical records of large health systems may serve as a valuable tool for nutritional surveillance among non low-income populations. PMID- 17406186 TI - Shouldn't we be making better use of the best available health measurement technologies? PMID- 17406187 TI - Systemic hypertension, headache, and ocular hemodynamics: a new hypothesis. AB - The association between systemic hypertension and headache remains controversial and its pathophysiologic basis is uncertain. A rather characteristic early morning pulsating headache is commonly seen in hypertensive patients, and a recent meta-analysis supports the link between these 2 entities. Epidemiologic evidence has paradoxically suggested a negative association between hypertension and headache. Unpredictable clinical association between severe hypertension and headache indicates that another cranial perfusion-related variable exerts a critical role. Neuroanatomically, head and neck pain primarily involves the ophthalmic division of the trigeminal nerve (V1). A link between systemic hypertension, pulsatile choroidal blood flow (CBF), and intraocular pressure (IOP) has been established. I propose that a trait ocular sympathetic hypofunction permits rapid episodic ocular choroidal overperfusion that stretches the ocular globe in the cohort of hypertensive patients with headache. Rapid distension of the pain-sensitive corneoscleral envelope can stimulate corneoscleral and iridial pain-sensitive V1 nerve endings and generate headache. Ocular tamponade function physiologically limits choroidal overperfusion. A higher basal IOP in some patients with moderate-to-severe hypertension may dampen pulsatile CBF and account for the negative epidemiologic link between sustained systemic hypertension and headache. Besides activation of the baroreceptor reflex, the association of hypalgesia with hypertension probably involves activation of the vasopressin-endorphin adaptive system consequent to mechanical stimulation of V1. The analogy between hypertensive headache and angle-closure glaucoma is rather limited because typical ocular and visual signs and symptoms of angle-closure glaucoma are not seen in hypertension-related headache. Hypertensive crises, including those associated with pheochromocytoma, are not accompanied by attacks of angle-closure glaucoma. Glaucoma is not associated with ocular choroidal congestion, but with reduced pulsatile CBF. The predisposition to develop angle-closure glaucoma is theoretically not associated with ocular autonomic hypofunction and should be conceptually dissociated from this hypothesis. The hypothesis can be evaluated by establishing significant circadian elevations of blood pressure, including nondipping nighttime pattern as well as circadian and periheadache measurements of IOP in patients with attacks of hypertension-related headache. PMID- 17406188 TI - Self management of fatal familial insomnia. Part 1: what is FFI? AB - CONTEXT: Fatal familial insomnia (FFI) is a genetically transmitted neurodegenerative prion disease that incurs great suffering and has neither a treatment nor a cure. The clinical literature is devoid of management plans (other than palliative). Part 1 of this article reviews the sparse literature about FFI, including case descriptions. Part 2 of this paper describes the efforts of 1 patient (with the rapid-course Met-Met subtype) to contend with his devastating symptoms and improve the quality of his life. DESIGN: Interventions were based on the premise that some symptoms may be secondary to insomnia and not a direct result of the disease itself. Strategies (derived by trial and error) were devised to induce sleep and increase alertness. Interventions included vitamin supplementation, narcoleptics, anesthesia, stimulants, sensory deprivation, exercise, light entrainment, growth hormone, and electroconvulsive therapy. RESULTS: The patient exceeded the average survival time by nearly 1 year, and during this time (when most patients are totally incapacitated), he was able to write a book and to successfully drive hundreds of miles. CONCLUSION: Methods to induce sleep may extend and enhance life during the disease, although they do not prevent death. It is hoped that some of his methods might inspire further clinical studies. PMID- 17406189 TI - Self-management of fatal familial insomnia. Part 2: case report. AB - CONTEXT: Fatal familial insomnia (FFI) is a genetically transmitted neurodegenerative prion disease that incurs great suffering and has neither a treatment nor cure. The clinical literature is devoid of management plans (other than palliative). Part 1 of this article reviews the sparse literature about FFI, including case descriptions. Part 2 describes the efforts of one patient (with the rapid-course Met-Met subtype) who contended with his devastating symptoms and improved the quality of his life. DESIGN: Interventions were based on the premise that some symptoms may be secondary to insomnia and not a direct result of the disease itself. Strategies (derived by trial and error) were devised to induce sleep and increase alertness. Interventions included vitamin supplementation, narcoleptics, anesthesia, stimulants, sensory deprivation, exercise, light entrainment, growth hormone, and electroconvulsive therapy (ECT). RESULTS: The patient exceeded the average survival time by nearly 1 year, and during this time (when most patients are totally incapacitated), he was able to write a book and to successfully drive hundreds of miles. CONCLUSION: Methods to induce sleep may extend and enhance life during the disease course, although they do not prevent death. It is hoped that some of his methods will inspire further clinical studies. PMID- 17406190 TI - The store front, Fountain House, and the rehabilitation alliance. PMID- 17406191 TI - Pursuing Ponce's dream: enabling the "square-wave life curve". PMID- 17406192 TI - Health information technology: is the time past for "watchful waiting?". PMID- 17406193 TI - Lyme neuroborreliosis presenting as the syndrome of inappropriate antidiuretic hormone secretion. AB - We describe a case of a patient presenting with the syndrome of inappropriate hormone secretion (SIADH) caused by Lyme neuroborreliosis. PMID- 17406194 TI - The problem with gastric varices. PMID- 17406195 TI - Hope in the neonatal intensive care nursery: values, ethics, and the injury of continued existence. PMID- 17406196 TI - Our patients are sicker--posting grades on the classroom, office, or hospital door. PMID- 17406197 TI - Ten guidelines for culturally and linguistically appropriate care. PMID- 17406198 TI - Is "first do no harm" a lost concept in medical education. PMID- 17406199 TI - Attitudes toward the autopsy--an 8-state survey. AB - CONTEXT: National autopsy rates have declined for several decades, and the reasons for such decline remain contentious. OBJECTIVE: To elicit the opinions of one group of crucial decision makers as to the reasons for this decline and possible modes of reversal. DESIGN: A 2-part survey, composed of multiple choice questions and questions requesting specific data on autopsy rates and costs. SETTING: Illinois, Iowa, Louisiana, Minnesota, Nebraska, North Dakota, South Dakota, and Wisconsin. PARTICIPANTS: Hospital administrators within the 8 states. MAIN OUTCOME MEASURES: Six-point survey scale relating to reasons for autopsy decline and possible remedial measures, as well as estimates of autopsy rates and costs. RESULTS: The response rate was 43% and the median autopsy rate was 2.4% (mean 6.1%). The median cost of autopsy was estimated at $852 (mean $1275). Larger hospitals were associated with higher autopsy rates than smaller hospitals (9.6% vs 4.0%), and teaching hospitals had a significantly higher autopsy rate than nonteaching institutions (11.4% vs 3.8%). Autopsy rates also varied by type of hospital control, with federal government hospitals having the highest autopsy rate at 15.1%. Sixty-six percent of all respondents agreed that current autopsy rates were adequate. Of the respondents, the highest percent (86%) agreed that improved diagnostics contributed to the decline in autopsies, and the highest percent (78%) agreed that direct payment to pathologists for autopsies under the physician fee schedule might lead to an increase in autopsies. CONCLUSIONS: Our data support the conclusion that the decline in autopsy performance is multifactorial, although the variable that dominates in this analysis is the contentious perception that improved diagnostic technology renders the autopsy redundant. The rate of autopsy is conditional, at least in part, on individual hospital characteristics such as large hospital size, teaching status, and federal ownership. Three underlying factors may explain these associations: resources, mission, and case mix. An important factor in declining autopsy rates appears to be the changing economic landscape, with its increased focus on cost control within both the public and private healthcare sectors. PMID- 17406200 TI - Hypertensive retinopathy associated with use of the ephedra-free weight-loss herbal supplement Hydroxycut. AB - The use of performance-enhancing and weight-loss supplements is prevalent in the United States, and over the past decade, there has been growing concern with regard to the safety and efficacy of these products. It is well documented that ephedra-based products are associated with adverse reactions, including serious cardiovascular and neurologic injuries. With new restrictions placed on such products, companies are now marketing caffeine-based ephedra-free herbal supplements. Less is known about the potential side effects of these products. We present the case of a 42-year-old, previously healthy man who developed malignant hypertension and hypertensive retinopathy while taking Hydroxycut, a caffeine based ephedra-free supplement. To our knowledge, this is the first documented case of hypertensive retinopathy associated with the use of Hydroxycut. Given the lack of investigative studies in regard to their safety and efficacy, judicious care should be taken with the use of all herbal supplements, including those designated as ephedra-free. PMID- 17406201 TI - Nothing to hide: transparency in healthcare. PMID- 17406202 TI - Ethics and anomalous (alternative) medical practices. PMID- 17406203 TI - Construction of semi-randomized gene libraries with weighted oligonucleotide synthesis and PCR. AB - Randomized gene libraries may be constructed and screened to find novel candidates with particular functions, and the applications can range widely, from protein engineering to selecting new microRNAs. Here we describe a technique to construct gene libraries using semi-randomized weighted oligonucleotide synthesis and end-to-end ligation. This method makes it possible to search the combinatorial space around a particular nucleotide sequence for a greater number of positions than is possible with fully randomized oligonucleotides. As an alternative to full cassette construction, library mutations can also be introduced through 'round-the-world PCR' approaches. Construction of a randomized gene cassette and cloning can typically be achieved in 2 weeks. Therefore, these are rapid and convenient methods to generate successive generations of libraries for iterative selection and optimization. PMID- 17406204 TI - Live-cell assay to detect antigen-specific CD4+ T-cell responses by CD154 expression. AB - This protocol details a method to identify CD4+ T cells that respond to antigens. The method relies on detection of CD154, a costimulatory cell surface protein that is expressed by CD4+ T cells upon activation, and can be used to purify live CD4+ T cells of diverse function. To detect CD154, fluorescently labeled antibodies are cultured with cell samples, peptides (or whole antigens) and monensin during a 6- to 24-h stimulation period. (Note that the assay is not compatible with brefeldin A.) After stimulation, cells are stained with any other antibodies of interest and then are analyzed by flow cytometry or purified by cell sorting. Unlike other assays, this method allows simultaneous assessment of other cell phenotypes or functions, is compatible with downstream RNA-based assays and preserves cell viability. This protocol can be completed in 9 h. PMID- 17406205 TI - T-maze alternation in the rodent. AB - This protocol details a method for using a T-maze to assess the cognitive ability of rodents. The T-maze is an elevated or enclosed apparatus in the form of a T placed horizontally. Animals are started from the base of the T and allowed to choose one of the goal arms abutting the other end of the stem. If two trials are given in quick succession, on the second trial the rodent tends to choose the arm not visited before, reflecting memory of the first choice. This is called 'spontaneous alternation'. This tendency can be reinforced by making the animal hungry and rewarding it with a preferred food if it alternates. Both spontaneous and rewarded alternation are very sensitive to dysfunction of the hippocampus, but other brain structures are also involved. Each trial should be completed in under 2 min, but the total number of trials required will vary according to statistical and scientific requirements. PMID- 17406206 TI - Appetitive position discrimination in the T-maze. AB - This protocol details a method to perform appetitively motivated tasks in rodents to test cognitive ability. When testing cognition in animals, the simplest paradigms can potentially yield quick results with minimal investment from the experimenter. Although appetitively motivated tasks are generally learnt more slowly than aversively motivated ones, they may be essential for distinguishing the effects of a treatment on learning from its effects on aversive motivation per se. For example, if a treatment improves learning in both types of paradigm, this is better evidence that it affects cognition rather than sensorimotor processes. Rats and mice easily learn position discriminations in a T-maze, especially if multiple cues, such as different objects and floor textures in the goal arms, are provided. To start, the rodent is placed in the maze and it chooses an arm. This Trial 1, however, is the only one on which this arm will be rewarded. From now on, it must always choose the other arm. The rule is simple: for example, always turn left into the arm with diagonal black stripes on the walls and gravel glued to the floor. High levels of correct responding can be achieved within 20-40 trials. The test may therefore be particularly useful with animals of low cognitive ability, such as transgenic mice derived from some 129 or SJL strains. Once the animals are habituated, each trial should take approximately 1 min. Thus, to test ten animals for 40 trials would take around 7 h. PMID- 17406207 TI - Tricine-SDS-PAGE. AB - Tricine-SDS-PAGE is commonly used to separate proteins in the mass range 1-100 kDa. It is the preferred electrophoretic system for the resolution of proteins smaller than 30 kDa. The concentrations of acrylamide used in the gels are lower than in other electrophoretic systems. These lower concentrations facilitate electroblotting, which is particularly crucial for hydrophobic proteins. Tricine SDS-PAGE is also used preferentially for doubled SDS-PAGE (dSDS-PAGE), a proteomic tool used to isolate extremely hydrophobic proteins for mass spectrometric identification, and it offers advantages for resolution of the second dimension after blue-native PAGE (BN-PAGE) and clear-native PAGE (CN PAGE). Here I describe a protocol for Tricine-SDS-PAGE, which includes efficient methods for Coomassie blue or silver staining and electroblotting, thereby increasing the versatility of the approach. This protocol can be completed in 1-2 d. PMID- 17406208 TI - The comet assay: a method to measure DNA damage in individual cells. AB - We present a procedure for the comet assay, a gel electrophoresis-based method that can be used to measure DNA damage in individual eukaryotic cells. It is versatile, relatively simple to perform and sensitive. Although most investigations make use of its ability to measure DNA single-strand breaks, modifications to the method allow detection of DNA double-strand breaks, cross links, base damage and apoptotic nuclei. The limit of sensitivity is approximately 50 strand breaks per diploid mammalian cell. DNA damage and its repair in single-cell suspensions prepared from yeast, protozoa, plants, invertebrates and mammals can also be studied using this assay. Originally developed to measure variation in DNA damage and repair capacity within a population of mammalian cells, applications of the comet assay now range from human and sentinel animal biomonitoring (e.g., DNA damage in earthworms crawling through toxic waste sites) to measurement of DNA damage in specific genomic sequences. This protocol can be completed in fewer than 24 h. PMID- 17406209 TI - Identifying DNA sequences recognized by a transcription factor using a bacterial one-hybrid system. AB - Bacterial-based interaction trap systems provide a powerful method to identify interacting macromolecules. When carried out in the context of a genetic selection, interacting pairs can be rapidly isolated from large combinatorial libraries. This technology has been adapted to allow the identification of DNA binding sequences for a transcription factor (TF) from a large randomized library. This procedure uses a library of randomized binding sites upstream of a cocistronic HIS3-URA3 reporter cassette. The URA3 reporter allows self-activating sequences to be removed from the library through counter-selection. The HIS3 reporter allows sequences that are recognized by a TF to be isolated from the library, where transcriptional activation is mediated by fusion of the TF to the alpha-subunit of RNA polymerase. This technology can be used to characterize monomeric, homodimeric and heterodimeric DNA-binding domains and, once a suitable library is constructed, binding sites can be identified in approximately 10 d. The bacterial one-hybrid system allows larger libraries to be searched than the corresponding yeast one-hybrid system and, unlike SELEX, it does not require purification of the TF(s). The complexity of the binding site libraries that can be searched using the bacterial system is, however, more limited than SELEX, and some eukaryotic factors may not express or fold efficiently in the bacterial system. PMID- 17406210 TI - In-gel stable isotope labeling for relative quantification using mass spectrometry. AB - Although differences in protein staining intensity can often be visualized by difference gel electrophoresis, abundant proteins can obscure less abundant proteins, and quantification of post-translational modifications is difficult. We present a protocol for quantifying changes in the abundance of a specific protein or changes in specific modifications of a protein using in-gel stable isotope labeling. In this protocol protein extracts from any source treated under two experimental conditions are resolved in two separate lanes by gel electrophoresis. Parallel gel regions of interest are reacted separately with either light or heavy isotope-labeled reagents, and the gel slices are then combined and digested with proteases. The resulting peptides are then analyzed by liquid chromatography/mass spectrometry (LC/MS) to determine relative abundance of light- and heavy-isotope lysine-containing peptide pairs and analyzed by LC/MS/MS for identification of sequence and modifications. This protocol should take approximately 24-26 h to complete, including the incubation time for proteolytic digestion. Additional time will be needed for data analysis and interpretation. PMID- 17406211 TI - Quantification of DNA methylation in electrofluidics chips (Bio-COBRA). AB - Alterations of normal gene expression patterns are a hallmark of human cancers. It is now clear that the dysregulation of epigenetic modifications of the DNA and surrounding histones contributes to aberrant gene silencing, thus being major participants not only in the progression but also the initiation of the disease phenotype. The best-studied epigenetic modification is DNA methylation, which converts cytosine to 5-methylcytosine. Aberrant hypermethylation of the promoter is frequently observed in cancer and is generally associated with gene silencing. Currently, accurate and reproducible quantification of DNA methylation remains challenging. Here, we describe Bio-COBRA, a modified protocol for Combined Bisulfite Restriction Analysis (COBRA), that incorporates an electrophoresis step in microfluidics chips. Microfluidics technology involves the handling of small amounts of liquid in miniaturized systems. In the life sciences, microfluidics usually entails the scaling down of at least one application, such as electrophoresis, to chip format, which often results in increased efficiency and reliability. Bio-COBRA provides a platform for the rapid and quantitative assessment of DNA methylation patterns in large sample sets. Its sensitivity and reproducibility also makes it a tool for the analysis of DNA methylation in clinical samples. The Bio-COBRA assay can be performed on 12 samples in less than 1 h. If the protocol is started at the DNA isolation step, however, approximately 48 h would be required to complete the entire procedure. PMID- 17406212 TI - Biomolecular screening of formylpeptide receptor ligands with a sensitive, quantitative, high-throughput flow cytometry platform. AB - The formylpeptide receptor (FPR) family of G protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. Here we describe a high-throughput flow cytometry screening approach that has successfully identified multiple families of previously unknown FPR ligands. The assay detects active structures that block the binding of a fluorescent ligand to membrane FPR of intact cells, thus detecting both agonists and antagonists. It is homogeneous in that assay reagents are added in sequence and the wells are subsequently analyzed without intervening wash steps. Microplate wells are routinely processed at a rate of 40 wells per minute, requiring a volume of only 2 microl to be sampled from each. This screening approach has recently been extended to identify a high-affinity, selective agonist for the intracellular estrogen-binding G protein-coupled receptor GPR30. With the development of appropriate assay reagents, it may be generally adaptable to a wide range of receptors. The total time required for the assay ranges between 1.5 and 2.5 h. The time required for flow cytometry analysis of a 96-well plate at the end of the procedure is less than 2.5 min. By comparison, manual processing of 96 samples will typically require 40-50 min, and a fast commercial automated sampler processes 96-well plates in less than 15 min, requiring the aspiration of 22 microl per sample for an analysis volume of 2 microl. PMID- 17406213 TI - Unrestrictive identification of post-translational modifications through peptide mass spectrometry. AB - Proteins are post-translationally modified in vivo as part of cellular regulation and signaling, and undergo further chemical modifications during laboratory processing. Even relatively simple protein samples may carry a wide range of modifications. Peptide tandem mass spectrometry provides a way to study these events. We present a protocol for computational identification of post translational modifications (PTMs) and the sites where they occur. The protocol performs an unrestrictive search, and requires no prior knowledge of what modifications are present in the sample. We present a largely automated procedure for PTM discovery, and provide a guide for analysis of PTM annotations. This protocol requires you to type out several commands, so you may wish to enlist the help of a colleague familiar with the computer's command-line interface. A typical MS run of up to 25,000 scans can be searched and analyzed in 3 h. PMID- 17406214 TI - Labeling of immune cells for in vivo imaging using magnetofluorescent nanoparticles. AB - Observation of immune and stem cells in their native microenvironments requires the development of imaging agents to allow their in vivo tracking. We describe here the synthesis of magnetofluorescent nanoparticles for cell labeling in vitro and for multimodality imaging of administered cells in vivo. MION-47, a prototype monocrystalline iron oxide nanoparticle, was first converted to an intermediate bearing a fluorochrome and amine groups, then reacted with either HIV-Tat peptide or protamine to yield a nanoparticle with membrane-translocating properties. We describe how to assess optimal cell labeling with tests of cell phenotype and function. Synthesis of magnetofluorescent nanoparticles and cell-labeling optimization can be realized in 48 h, whereas nanoparticle uptakes and retention studies may generally take up to 120 h. Labeled cells can be detected by magnetic resonance imaging, fluorescence reflectance imaging, fluorescence-mediated tomography, confocal microscopy and flow cytometry, and can be purified based on their fluorescent or magnetic properties. The present protocol focuses on T-cell labeling but can be used for labeling a variety of circulating cells. PMID- 17406215 TI - Cysteine engineering of polyproteins for single-molecule force spectroscopy. AB - Single-molecule methods such as force spectroscopy give experimental access to the mechanical properties of protein molecules. So far, owing to the limitations of recombinant construction of polyproteins, experimental access has been limited to mostly the N-to-C terminal direction of force application. This protocol gives a fast and simple alternative to current recombinant strategies for preparing polyproteins. We describe in detail the method to construct polyproteins with precisely controlled linkage topologies, based on the pairwise introduction of cysteines into protein structure and subsequent polymerization in solution. Stretching such constructed polyproteins in an atomic force microscope allows mechanical force application to a single protein structure via two precisely controlled amino acid positions in the functional three-dimensional protein structure. The capability for site-directed force application can provide valuable information about both protein structure and directional protein mechanics. This protocol should be applicable to almost any protein that can be point mutated. Given correct setup of all necessary reagents, this protocol can be accomplished in fewer than 10 d. PMID- 17406216 TI - The gelatin sponge-chorioallantoic membrane assay. AB - Here we present a method for the quantification of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane (CAM) based on the implantation of a gelatin sponge on the top of the growing CAM on day 8 of development. After implantation, the sponge is treated with a stimulator of blood vessel formation in the absence or presence of an angiogenesis inhibitor. On day 12, blood vessels that are growing into the sponge are counted at macroscopic and microscopic levels. The estimated timeline for carrying out this protocol is 10 d. The presence of a vascular network in the CAM requires a careful analysis to distinguish new capillaries from pre-existing ones. This limitation does not occur in the avascular cornea assay, which may also take advantage of different genetic backgrounds when carried out in transgenic or knockout mice. Nevertheless, the gelatin sponge-CAM assay is simple, inexpensive and suitable for large-scale screening. PMID- 17406217 TI - Reverse MAPPIT detects disruptors of protein-protein interactions in human cells. AB - Reverse mammalian protein-protein interaction trap (MAPPIT) is a mammalian reverse two-hybrid technology. The method is adapted from the forward MAPPIT technique, a two-hybrid complementation system in which the interaction between a bait-fusion protein and a prey-fusion protein restores ligand-dependent cytokine receptor signaling. In the reverse mode described in detail here, a positive readout is generated on disruption of the designated protein-protein interactions. Reverse MAPPIT functions in intact human cells, facilitating simultaneous analysis of disruption, toxicity and permeability of the tested compounds, making it particularly suitable for screening for molecules that target therapeutically interesting protein-protein interactions or for mapping the interaction interface between proteins. The total handling time of a typical reverse MAPPIT experiment is approximately 9 h and is spread over 4-5 d. PMID- 17406218 TI - Immunocytochemical techniques for whole-mount in situ protein localization in plants. AB - As the field of plant molecular biology is swiftly advancing, a need has been created for methods that allow rapid and reliable in situ localization of proteins in plant cells. Here we describe a whole-mount 'immunolocalization' technique for various plant tissues, including roots, hypocotyls, cotyledons, young primary leaves and embryos of Arabidopsis thaliana and other species. The detailed protocol, recommended controls and troubleshooting are presented, along with examples of applications. The protocol consists of five main procedures: tissue fixation, tissue permeation, blocking, primary and secondary antibody incubation. Notably, the first procedure (tissue fixation) includes several steps (4-12) that are absolutely necessary for protein localization in hypocotyls, cotyledons and young primary leaves but should be omitted for other tissues. The protocol is usually done in 3 days, but could also be completed in 2 days. PMID- 17406219 TI - Immunocytochemical technique for protein localization in sections of plant tissues. AB - There is a growing demand for methods that allow rapid and reliable in situ localization of proteins in plant cells. The immunocytochemistry protocol presented here can be used routinely to observe protein localization patterns in tissue sections of various plant species. This protocol is especially suitable for plant species with more-complex tissue architecture (such as maize, Zea mays), which makes it difficult to use an easier whole-mount procedure for protein localization. To facilitate the antibody-antigen reaction, it is necessary to include a wax-embedding and tissue-sectioning step. The protocol consists of the following procedures: chemical fixation of tissue, dehydration, wax embedding, sectioning, dewaxing, rehydration, blocking and antibody incubation. The detailed protocol, recommended controls and troubleshooting are presented here, along with examples of applications. PMID- 17406220 TI - Radiolabeling of HYNIC-annexin V with technetium-99m for in vivo imaging of apoptosis. AB - Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of the pathophysiology and treatment of these and numerous other human disorders. Here we describe the most used method for labeling annexin V, a protein with a high affinity for apoptotic cells in vitro, with technetium-99m (99mTc) as a radionuclide imaging agent that can localize and non-invasively quantify apoptosis in vivo when coupled with single-photon emission tomography. In this method, annexin V is first attached to the bifunctional chelator molecule hydrazino nicotinate (HYNIC). Once prepared, HYNIC-annexin V can be labeled with 99mTc, a widely available gamma-radiation-emitting radionuclide, for intravenous injection in as little as 30 min without the need for specialized reagents or equipment. PMID- 17406221 TI - Azido ruthenium: a new photoreactive probe for calcium-binding proteins. AB - Ca2+, involved in almost all processes of cell life, mediates its activity through reversible interaction with specific binding sites in proteins. Although several Ca2+-dependent activities are known, many of the proteins responsible remain unidentified. Here we describe the synthesis, purification, characterization and potential uses of a new Ca2+-like reagent, azido ruthenium (AzRu), which can be photoactivated. AzRu strongly inhibits Ca2+-dependent activities. AzRu can be used to probe proteins in solution or embedded in membranes. AzRu has no effect on Ca2+-independent or Mg2+-dependent activity. After exposure to ultraviolet irradiation, AzRu binds covalently and specifically to Ca2+-binding proteins, thus providing a new approach for identifying and purifying Ca2+-binding proteins, for characterizing their Ca2+-binding sites and for exploring previously unknown Ca2+-dependent processes. In this protocol we also include a description of the preparation of [103Ru]AzRu, which can be used for labeling Ca2+-binding sites in proteins and identifying previously unknown Ca2+-binding proteins. The preparation of AzRu takes approximately 2-3 days. PMID- 17406222 TI - Burrowing in rodents: a sensitive method for detecting behavioral dysfunction. AB - Virtually all rodents display burrowing behavior, yet measurement of this behavior has not yet been standardized or formalized. Previously, parameters such as the latency to burrow and the complexity of the burrow systems in substrate filled boxes in the laboratory or naturalistic outdoor environments have been assessed. We describe here a simple protocol that can quantitatively measure burrowing in laboratory rodents, using a simple apparatus that can be placed in the home cage. The test is very cheap to run and requires minimal experimenter training, yet seems sensitive to a variety of treatments, such as the early stages of prion disease in mice, mouse strain differences, lesions of the hippocampus and prefrontal cortex in mice, also effects of lipopolysaccharide and IL-1beta in rats. Other species such as hamsters, gerbils and Egyptian spiny mice also burrow in this apparatus, and with suitable size modification probably almost any burrowing animal could be tested in it. The simplicity, sensitivity and robustness of burrowing make it ideal for assessing genetically modified animals, which in most cases would be mice. The test is run from late afternoon until the next morning, but only two measurements need to be taken. PMID- 17406223 TI - Digging and marble burying in mice: simple methods for in vivo identification of biological impacts. AB - Mice exhibit various species-typical behaviors such as digging and burrowing. They dig in the ground to find food, to hoard food, to create a refuge from predators or cold and to make a safe nursery area for the young. In the laboratory, mice dig vigorously in deep bedding such as wood chips. This behavior is sensitive to strain differences and drugs. For example, the effects of anxiolytics and 5-HT-active compounds, including those used clinically for obsessive-compulsive disorder (OCD), can be detected. Digging can be quantified by manual timing. Alternatively, the bedding can be covered with glass marbles and the number buried can be counted after a set time. These behaviors can be assessed using very little specialized equipment, and results can be obtained from ten animals in about an hour. Species-typical behaviors may be sensitive to a wide variety of treatments, and their simplicity and ability to yield robust quantitative data might be particularly useful in assessing genetically modified mice, even in laboratories not primarily oriented to behavioral work. PMID- 17406224 TI - Production of cloned mice by somatic cell nuclear transfer. AB - Although it has now been 10 years since the first cloned mammals were generated from somatic cells using nuclear transfer (NT), the success rate for producing live offspring by cloning remains < 5%. Nevertheless, the techniques have potential as important tools for future research in basic biology. We have been able to develop a stable NT method in the mouse, in which donor nuclei are directly injected into the oocyte using a piezo-actuated micromanipulator. Although manipulation of the piezo unit is complex, once mastered it is of great help not only in NT experiments but also in almost all other forms of micromanipulation. In addition to this technique, embryonic stem (ES) cell lines established from somatic cell nuclei by NT can be generated relatively easily from a variety of mouse genotypes and cell types. Such NT-ES cells can be used not only for experimental models of human therapeutic cloning but also as a backup of the donor cell's genome. Our most recent protocols for mouse cloning, as described here, will allow the production of cloned mice in > or = 3 months. PMID- 17406225 TI - Quantitative proteome analysis using isotope-coded affinity tags and mass spectrometry. AB - A main objective of proteomics research is to systematically identify and quantify proteins in a given proteome (cells, subcellular fractions, protein complexes, tissues or body fluids). Protein labeling with isotope-coded affinity tags (ICAT) followed by tandem mass spectrometry allows sequence identification and accurate quantification of proteins in complex mixtures, and has been applied to the analysis of global protein expression changes, protein changes in subcellular fractions, components of protein complexes, protein secretion and body fluids. This protocol describes protein-sample labeling with ICAT reagents, chromatographic fractionation of the ICAT-labeled tryptic peptides, and protein identification and quantification using tandem mass spectrometry. The method is suitable for both large-scale analysis of complex samples including whole proteomes and small-scale analysis of subproteomes, and allows quantitative analysis of proteins, including those that are difficult to analyze by gel-based proteomics technology. PMID- 17406226 TI - In-cell NMR for protein-protein interactions (STINT-NMR). AB - We describe an in-cell NMR-based method for mapping the structural interactions (STINT-NMR) that underlie protein-protein complex formation. This method entails sequentially expressing two (or more) proteins within a single bacterial cell in a time-controlled manner and monitoring their interactions using in-cell NMR spectroscopy. The resulting NMR data provide a complete titration of the interaction and define structural details of the interacting surfaces at atomic resolution. Unlike the case where interacting proteins are simultaneously overexpressed in the labeled medium, in STINT-NMR the spectral complexity is minimized because only the target protein is labeled with NMR-active nuclei, which leaves the interactor protein(s) cryptic. This method can be combined with genetic and molecular screens to provide a structural foundation for proteomic studies. The protocol takes 4 d from the initial transformation of the bacterial cells to the acquisition of the NMR spectra. PMID- 17406227 TI - mini-Tn7 insertion in bacteria with single attTn7 sites: example Pseudomonas aeruginosa. AB - Broad host-range mini-Tn7 vectors facilitate integration of single-copy genes into bacterial chromosomes at a neutral, naturally evolved site. Here we present a protocol for employing the mini-Tn7 system in bacteria with single attTn7 sites, using the example Pseudomonas aeruginosa. The procedure involves, first, cloning of the genes of interest into an appropriate mini-Tn7 vector; second, co transfer of the recombinant mini-Tn7 vector and a helper plasmid encoding the Tn7 site-specific transposition pathway into P. aeruginosa by either transformation or conjugation, followed by selection of insertion-containing strains; third, PCR verification of mini-Tn7 insertions; and last, optional Flp-mediated excision of the antibiotic-resistance selection marker present on the chromosomally integrated mini-Tn7 element. From start to verification of the insertion events, the procedure takes as little as 4 d and is very efficient, yielding several thousand transformants per microgram of input DNA or conjugation mixture. In contrast to existing chromosome integration systems, which are mostly based on species-specific phage or more-or-less randomly integrating transposons, the mini Tn7 system is characterized by its ready adaptability to various bacterial hosts, its site specificity and its efficiency. Vectors have been developed for gene complementation, construction of gene fusions, regulated gene expression and reporter gene tagging. PMID- 17406228 TI - mini-Tn7 insertion in bacteria with multiple glmS-linked attTn7 sites: example Burkholderia mallei ATCC 23344. AB - The mini-Tn7 vectors are universally applicable in gram-negative bacteria and thereby facilitate the manipulation of many organisms for which few genetic systems are available. These vectors, when provided with only the Tn7 site specific transposition machinery, insert site and orientation specifically in the bacterial chromosome at an attTn7 site downstream of the essential glmS gene. A few bacteria, including Burkholderia spp., contain multiple glmS genes and therefore several attTn7 sites. Here we provide a protocol for application of the mini-Tn7 system in B. mallei as an example of bacteria with multiple glmS sites. The procedure involves, first, cloning of the genes of interest into an appropriate mini-Tn7 vector; second, co-transfer of the recombinant mini-Tn7 vector and a helper plasmid encoding the Tn7 site-specific transposition pathway into B. mallei by conjugation, followed by selection of insertion-containing strains; and last, PCR verification of mini-Tn7 insertions. B. mallei possesses two glmS genes on chromosome 1 and Tn7 transposes to both sites, although transposition to attTn7-1 associated with glmS1 occurs in more than 90% of the clones examined. Transposition is efficient and the whole procedure from start to verification of insertion events can be done in less than 5 d. This first chromosome integration system in B. mallei provides an important contribution to the genetic tools emerging for Burkholderia spp. Vectors are available for gene complementation and expression, and gene fusion analyses. PMID- 17406229 TI - mini-Tn7 insertion in bacteria with secondary, non-glmS-linked attTn7 sites: example Proteus mirabilis HI4320. AB - We previously constructed a series of mini-Tn7 chromosome integration vectors that, when provided only with the site-specific transposition machinery, generally transpose to a naturally evolved, neutral attTn7 site that is located 25-bp downstream of the glmS gene. Here we provide a protocol for application of the mini-Tn7 system in Proteus mirabilis as an example of a bacterium with a secondary attTn7 site that is not linked to glmS but, in this case, located in the carAB operon. The procedure involves, first, cloning of the genes of interest into an appropriate mini-Tn7 vector; second, co-transfer of the recombinant mini Tn7 vector and a helper plasmid encoding the Tn7 site-specific transposition pathway into P. mirabilis by transformation, followed by selection of insertion containing strains; third, PCR verification of mini-Tn7 insertions; and last, optional Flp-mediated excision of the antibiotic-resistance selection marker present on the chromosomally integrated mini-Tn7 element. When transposon containing cells are selected on rich medium, insertions occur at both attTn7 sites with equal efficiency and frequency. Because carA mutants are arginine and pyrimidine auxotrophs, single-site insertions at the glmS attTn7 sites can be obtained by selection on minimal medium. From start to verification of the insertion events, the whole procedure takes 5 d. This chromosome integration system in P. mirabilis provides an important tool for animal and biofilm studies based on this bacterium. Vectors are available for gene complementation and expression, gene fusion analyses and tagging with a green fluorescent protein (GFP)-encoding reporter gene. PMID- 17406230 TI - Protocol for the fast chromatin immunoprecipitation (ChIP) method. AB - Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations (ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein-DNA interactions in vivo and to estimate the density of proteins at specific sites genome-wide. We have introduced several improvements to the traditional ChIP assay, which simplify the procedure, greatly reducing the time and labor required to complete the assay. The simplicity of the method yields highly reproducible results. Our improvements facilitate the probing of multiple proteins in a single experiment, which allows for the simultaneous monitoring of many genomic events. This method is particularly useful in kinetic studies where multiple samples are processed at the same time. Starting with sheared chromatin, PCR-ready DNA can be isolated from 16-24 ChIP samples in 4-6 h using the fast method. PMID- 17406231 TI - Isothermal titration calorimetry to determine association constants for high affinity ligands. AB - An important goal in drug development is to engineer inhibitors and ligands that have high binding affinities for their target molecules. In optimizing these interactions, the precise determination of the binding affinity becomes progressively difficult once it approaches and surpasses the nanomolar level. Isothermal titration calorimetry (ITC) can be used to determine the complete binding thermodynamics of a ligand down to the picomolar range by using an experimental mode called displacement titration. In a displacement titration, the association constant of a high-affinity ligand that cannot be measured directly is artificially lowered to a measurable level by premixing the protein with a weaker competitive ligand. To perform this protocol, two titrations must be carried out: a direct titration of the weak ligand to the target macromolecule and a displacement titration of the high-affinity ligand to the weak ligand target macromolecule complex. This protocol takes approximately 5 h. PMID- 17406232 TI - In vivo protein biotinylation and sample preparation for the proteomic identification of organ- and disease-specific antigens accessible from the vasculature. AB - Targeted delivery of bioactive molecules to diseased organs or tissues by means of binding molecules specific to markers of diseases represents a promising area of pharmaceutical intervention. The availability of markers of pathology, ideally accessible from the vasculature, is crucial for such strategies. To this aim, here we present a protocol based on terminal perfusion of mice with a reactive ester derivate of biotin that enables the covalent modification of proteins readily accessible from the bloodstream. Biotinylated proteins from total organ or tissue extracts are (i) purified on streptavidin resin in the presence of strong detergents, (ii) digested on the resin and (iii) subjected to proteomic analysis. This technology is applicable to comparative proteomic investigations of differentially expressed, accessible proteins in numerous animal models having different physiological and pathological processes. PMID- 17406233 TI - A direct approach to quantification of the cellular uptake of cell-penetrating peptides using MALDI-TOF mass spectrometry. AB - This protocol allows the accurate quantification of cell-penetrating peptide (CPP) cellular uptake by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Quantification is based on the use of an internal standard with same chemical structure as the analyte but labeled with a stable isotope. The analyte and the standard can both be obtained by standard solid-phase peptide synthesis using commercially available amino acids. They are functionalized by biotin to allow their easy purification before MALDI-TOF MS analysis. The method allows determination of the amount of intact internalized peptide and the identification of potential intracellular digests. It can be used to simultaneously compare the uptake of several peptides, and can also be applied to the quantification of peptidic cargoes and the study of their intracellular stability. It is therefore a potent tool to study the mechanisms of CPPs internalization and to select new carriers for drug delivery. This protocol will take approximately 5 hours for the analysis of 12 samples (not including the time for cell incubation with peptides). PMID- 17406234 TI - Reconstruction of human mammary tissues in a mouse model. AB - Establishing a model system that more accurately recapitulates both normal and neoplastic breast epithelial development in rodents is central to studying human breast carcinogenesis. However, the inability of human breast epithelial cells to colonize mouse mammary fat pads is problematic. Considering that the human breast is a more fibrous tissue than is the adipose-rich stroma of the murine mammary gland, our group sought to bypass the effects of the rodent microenvironment through incorporation of human stromal fibroblasts. We have been successful in reproducibly recreating functionally normal breast tissues from reduction mammoplasty tissues, in what we term the human-in-mouse (HIM) model. Here we describe our relatively simple and inexpensive techniques for generating this orthotopic xenograft model. Whether the model is to be applied for understanding normal human breast development or tumorigenesis, investigators with minimal animal surgery skills, basic cell culture techniques and access to human breast tissue will be able to generate humanized mouse glands within 3 months. Clearing the mouse of its endogenous epithelium with subsequent stromal humanization takes 1 month. The subsequent implantation of co-mixed human epithelial cells and stromal cells occurs 2 weeks after humanization, so investigators should expect to observe the desired outgrowths 2 months afterward. As a whole, this model system has the potential to improve the understanding of crosstalk between tissue stroma and the epithelium as well as factors involved in breast stem cell biology tumor initiation and progression. PMID- 17406235 TI - High-throughput assay for determining specificity and affinity of protein-DNA binding interactions. AB - Limited information exists for the binding specificities of many important transcription factors. To address this, we have previously developed a microwell based assay for directly measuring the affinity of DNA-protein binding interactions. We describe here the detailed protocol for determining sequence specificities of DNA-binding proteins using this assay. The described method is rapid; after preparation of the reagents, the assay can be run in a single day, and its throughput can be increased further by automation. The method is quantitative but requires prior knowledge of one high-affinity binding site for the protein of interest. The protocol can be adapted for determining the effect of protein modifications and protein-protein interactions on DNA-binding specificity, and for engineering proteins with new DNA-binding specificities. In addition, the method is suitable for high-throughput screening to identify proteins or small molecules that modulate protein-DNA binding interactions. PMID- 17406236 TI - One-step analysis of protein complexes in microliters of cell lysate using indirect immunolabeling & fluorescence cross-correlation spectroscopy. AB - This protocol describes a 'mix-and-measure' procedure for the analysis of interactions of endogenous proteins in microliters of crude cell lysates. The proteins of interest are labeled by indirect immunofluorescence through simple addition of all primary and secondary antibodies to the lysate. Detection is based on fluorescence cross-correlation spectroscopy. Due to the minimal number of handling steps for sample preparation and the need of only microliters of sample, the approach enables the parallel and miniaturized analysis of protein protein interactions. No heterologous expression of proteins with detection tags is required. For this reason, the cellular processes leading to protein-protein interactions are not skewed by overexpression of individual components. This makes the approach particularly suitable for the parallel monitoring of interactions in signaling networks. Additionally, the approach enables the screening and titration of compounds interfering with interactions, especially for those interactions based on signaling-dependent post-translational modifications. This protocol can be completed in approximately 22 h, including a 16-h incubation phase. PMID- 17406237 TI - The counting Stroop: a cognitive interference task. AB - The counting Stroop is a validated Stroop task variant. Initially designed as a functional magnetic resonance imaging (fMRI) task for identifying brain regions subserving cognition and attention (dorsal anterior midcingulate cortex (daMCC) and dorsolateral prefrontal cortex (DLPFC)), it has been used to study cognition in healthy volunteers and to identify functional brain abnormalities in neuropsychiatric disorders, such as attention deficit hyperactivity disorder (ADHD). During the counting Stroop, subjects report by button-press the number of words (one to four) appearing on the screen, regardless of word meaning. Neutral word control trials contain single semantic category common animals (e.g., 'dog' written three times), while interference trials contain number words that are incongruent with the correct response (e.g., 'two' written four times). The counting Stroop can be completed in approximately 20 min per subject and can be used offline (behavioral performance) or with fMRI, positron emission tomography, event-related potentials, magnetoencephalography or intracranial recordings. PMID- 17406238 TI - Design and cloning of lentiviral vectors expressing small interfering RNAs. AB - RNA interference (RNAi) has emerged as a powerful technique to downregulate gene expression. The use of polIII promoters to express small hairpin RNAs (shRNAs), combined with the versatility and robustness of lentiviral vector-mediated gene delivery to a wide range of cell types offers the possibility of long-term downregulation of specific target genes both in vitro and in vivo. The use of silencing lentivectors allows for a rapid and convenient way of establishing cell lines (or transgenic mice) that stably express shRNAs for analysis of phenotypes produced by knockdown of a gene product. Here we present two possible protocols describing the design and cloning of silencing lentiviral vectors. These protocols can be completed in less than 3 weeks. PMID- 17406239 TI - Production and purification of lentiviral vectors. AB - Lentiviral vectors offer unique versatility and robustness as vehicles for gene delivery. They can transduce a wide range of cell types and integrate into the host genome in both dividing and post-mitotic cells, resulting in long-term expression of the transgene both in vitro and in vivo. This protocol describes how lentiviral vectors can be produced, purified and titrated. High titer suspensions can be routinely prepared with relative ease: a low-titer (10(6) viral particles/ml) unpurified preparation can be obtained 3 d after transfecting cells with lentiviral vector and packaging plasmids; a high-titer (10(9) viral particles/ml) purified preparation requires 2 more days. PMID- 17406240 TI - Preparation of aptamer-linked gold nanoparticle purple aggregates for colorimetric sensing of analytes. AB - Aptamers are single-stranded DNA or RNA molecules that can bind target molecules with high affinity and specificity. The conformation of an aptamer usually changes upon binding to its target analyte, and this property has been used in a wide variety of sensing applications, including detection based on fluorescence intensity, polarization, energy transfer, electrochemistry or color change. Colorimetric sensors are particularly important because they minimize or eliminate the necessity of using expensive and complicated instruments. Among the many colorimetric sensing strategies, metallic nanoparticle-based detection is desirable because of the high extinction coefficients and strong distance dependent optical properties of the nanoparticles. Here, we describe a protocol for the preparation of aptamer-linked gold nanoparticle purple aggregates that undergo fast disassembly into red dispersed nanoparticles upon binding of target analytes. This method has proved to be generally applicable for colorimetric sensing of a broad range of analytes. The time range for the entire protocol is approximately 5 d, including synthesis and functionalization of nanoparticles, preparation of nanoparticle aggregates and sensing. PMID- 17406241 TI - Colony filtration blotting for screening soluble expression in Escherichia coli. AB - We have developed a screen for detecting E. coli colonies that produce soluble recombinant target proteins at the colony level: the colony filtration (CoFi) blot. Colonies are transferred, induced and lysed on a filter membrane that can separate soluble proteins from inclusion bodies. Upon lysis, the soluble proteins diffuse through the filter membrane and are captured on a nitrocellulose membrane. The nitrocellulose membrane is incubated with antibodies or probes specific for the target protein and are then developed. In the resulting image, colonies expressing soluble protein can easily be identified. This protocol can be used to screen thousands of constructs in a matter of days, making it very suitable for expression libraries. The protocol is robust and flexible with regard to lysis conditions, induction temperatures and strains. The method requires only standard laboratory equipment and is based on immunochemicals used for western blotting. The following protocol describes the screening of a DNA library with detection done using chemiluminescence. Depending on induction temperature, the whole procedure can be performed in <2 d. PMID- 17406242 TI - Rapid functional assays of intracellular Ca2+ channels. AB - Functional assays of intracellular Ca2+ channels, such as the inositol 1,4,5 trisphosphate receptor (IP3R), have generally used 45Ca2+-flux assays, fluorescent indicators loaded within either the cytosol or the endoplasmic reticulum (ER) of single cells, or electrophysiological analyses. None of these methods is readily applicable to rapid, high-throughput quantitative analyses. Here we provide a detailed protocol for high-throughput functional analysis of native and recombinant IP3Rs. A low-affinity Ca2+ indicator (mag-fluo-4) trapped within the ER of permeabilized cells is shown to report changes in luminal free Ca2+ concentration reliably. An automated fluorescence plate reader allows rapid measurement of Ca2+ release from intracellular stores mediated by IP3R. The method can be readily adapted to other cell types or to the analysis of other intracellular Ca2+ channels. This protocol can be completed in 2-3 h. PMID- 17406243 TI - COBRA: combined binary ratio labeling of nucleic-acid probes for multi-color fluorescence in situ hybridization karyotyping. AB - Combined binary ratio labeling (COBRA) is designed to increase the multiplicity of fluorescence in situ hybridization (FISH)--i.e., the number of targets that can be distinguished simultaneously. In principle, chemical (ULS), enzymatic (nick translation or random priming) or PCR-based labeling procedures of probes can be used. The method was originally designed to label chromosome-painting probes, but has also been used for probe sets specific for subtelomeric regions. COBRA imaging requires a digital fluorescence microscope equipped for sequential excitation and recording of color images. Staining of all 24 human chromosomes is accomplished with only four fluorochromes, compared with five for methods based on combinatorial labeling. The COBRA procedure takes approximately 6 h laboratory work, 2-3 d incubation and a few hours imaging. The method is routinely applied in research (cultured cells from human or mouse origin) or to support clinical diagnosis, such as postnatal and perinatal genetic testing and in solid tumors. PMID- 17406244 TI - The fluorescence protease protection (FPP) assay to determine protein localization and membrane topology. AB - Correct localization and topology are crucial for the cellular function of a protein. To determine the topology of membrane proteins, a new technique, called the fluorescence protease protection (FPP) assay, can be applied. This assay uses the restricted proteolytic digestibility of GFP-tagged transmembrane proteins to indicate their intramembrane orientation. The sole requirements for FPP are the expression of GFP fusion proteins and the selective permeabilization of the plasma membrane, which permits a wide range of cell types and organelles to be investigated. The FPP assay can be carried out in a straightforward manner to obtain reliable results within minutes. Here we provide a step-by-step protocol for the assay. As an example, we use FPP to determine which terminus of an endoplasmic reticulum (ER) transmembrane protein is lumenal and which one is facing the cytosol. PMID- 17406245 TI - Site-specific protein labeling by Sfp phosphopantetheinyl transferase. AB - Sfp phosphopantetheinyl transferase covalently attaches small-molecule probes including biotin and various organic fluorophores to a specific serine residue in the peptidyl carrier protein (PCP) or a short 11-residue peptide tag ybbR through a phosphopantetheinyl linker. We describe here a protocol for site-specific protein labeling by Sfp-catalyzed protein post-translational modification that includes (i) expression and purification of Sfp, (ii) synthesis of small-molecule probe-CoA conjugates, (iii) construction of target protein fusions with PCP or the ybbR tag, (iv) labeling PCP- or ybbR-tagged target protein fusions in cell lysates and on live cell surfaces and (v) imaging fluorophore-labeled cell surface receptors by fluorescence microscopy. To follow this protocol, we advise that you allow 3 d for the expression and purification of Sfp phosphopantetheinyl transferase, 1 d for the synthesis and purification of the small-molecule probe CoA conjugates as the substrates of Sfp, 3 d for the cloning of target protein genes as fusions to the PCP or the ybbR tag in the appropriate plasmids and another 3 d for transfecting cell lines with the plasmids and the expression of PCP- or ybbR-tagged proteins. Labeling of the PCP- or the ybbR-tagged proteins in cell lysates or on cell surfaces should require only 15-30 min. PMID- 17406246 TI - Rapid generation of knockdown transgenic mice by silencing lentiviral vectors. AB - Lentiviral vectors are potent gene delivery vehicles that enable stable expression of transgenes in both dividing and postmitotic cells, including preimplantation embryos. We have developed lentiviral vectors carrying silencing cassettes consisting of an RNA polymerase III promoter expressing short hairpin RNAs. Transgenic mice can be generated rapidly by transduction of early embryos with lentiviral silencing vectors, resulting in mice with downregulated target genes. We describe two alternative early embryo transduction protocols (removal of zona pellucida and subzonal microinjection). These methodologies offer the possibility of large-scale generation of knockdown transgenic mice for functional genomic studies and enable the production of transgenic mice in 7 weeks. PMID- 17406247 TI - The emotional counting Stroop: a task for assessing emotional interference during brain imaging. AB - The emotional counting Stroop (ecStroop) is an emotional variant of the counting Stroop. Both of these tasks require a motor response instead of a spoken response for the purpose of minimizing head movement during functional MRI (fMRI). During this task, subjects report, by button press, the number of words (1-4) that appear on a screen, regardless of word meaning. Neutral word-control trials contain common words (e.g., 'cabinet' written three times), while interference trials contain emotional words (e.g., 'murder' written three times). The degree to which this task represents a true 'Stroop' interference task, in the sense that emotional words will increase motor-response times compared with neutral words, depends upon the subjects of the study and the words that are presented. Much research on the emotional Stroop task demonstrates that interference effects are observed in psychopathological groups in response to words that are specific to their disorder, and in normal subjects when the words are related to current concerns endorsed by them. The ecStroop task described here will produce reaction time-interference effects that are comparable to the traditional color-naming emotional Stroop. This protocol can be completed in approximately 20 min per subject. The protocol described here employs neutral words and emotional words that include general-negative words, as well as words specific to combat-related trauma. However, this protocol is amenable to any emotional word lists. PMID- 17406248 TI - The Dimensional Change Card Sort (DCCS): a method of assessing executive function in children. AB - The dimensional change card sort (DCCS) is an easily administered and widely used measure of executive function that is suitable for use with participants across a wide range of ages. In the standard version, children are required to sort a series of bivalent test cards, first according to one dimension (e.g., color), and then according to the other (e.g., shape). Most 3-year-olds perseverate during the post-switch phase, exhibiting a pattern of inflexibility similar to that seen in patients with prefrontal cortical damage. By 5 years of age, most children switch when instructed to do so. Performance on the DCCS provides an index of the development of executive function, and it is impaired in children with disorders such as attention-deficit/hyperactivity disorder (ADHD) and autism. We describe the protocol for the standard version (duration = 5 min) and the more challenging border version (duration = 5 min), which may be used with children as old as 7 years. PMID- 17406249 TI - RIP-Chip: the isolation and identification of mRNAs, microRNAs and protein components of ribonucleoprotein complexes from cell extracts. AB - RNA targets of multitargeted RNA-binding proteins (RBPs) can be studied by various methods including mobility shift assays, iterative in vitro selection techniques and computational approaches. These techniques, however, cannot be used to identify the cellular context within which mRNAs associate, nor can they be used to elucidate the dynamic composition of RNAs in ribonucleoprotein (RNP) complexes in response to physiological stimuli. But by combining biochemical and genomics procedures to isolate and identify RNAs associated with RNA-binding proteins, information regarding RNA-protein and RNA-RNA interactions can be examined more directly within a cellular context. Several protocols--including the yeast three-hybrid system and immunoprecipitations that use physical or chemical cross-linking--have been developed to address this issue. Cross-linking procedures in general, however, are limited by inefficiency and sequence biases. The approach outlined here, termed RNP immunoprecipitation-microarray (RIP-Chip), allows the identification of discrete subsets of RNAs associated with multi targeted RNA-binding proteins and provides information regarding changes in the intracellular composition of mRNPs in response to physical, chemical or developmental inducements of living systems. Thus, RIP-Chip can be used to identify subsets of RNAs that have related functions and are potentially co regulated, as well as proteins that are associated with them in RNP complexes. Using RIP-Chip, the identification and/or quantification of RNAs in RNP complexes can be accomplished within a few hours or days depending on the RNA detection method used. PMID- 17406250 TI - The Multi-Source Interference Task: an fMRI task that reliably activates the cingulo-frontal-parietal cognitive/attention network. AB - In this protocol we describe how to perform the Multi-Source Interference Task (MSIT), a validated functional magnetic resonance imaging (fMRI) task that reliably and robustly activates the cingulo-frontal-parietal cognitive/attention network (CFP network) within individual subjects. The MSIT can be used to (i) identify the cognitive/attention network in normal volunteers and (ii) test its integrity in people with neuropsychiatric disorders. It is simple to perform, can be completed in less than 15 min and is not language specific, making it appropriate for children, adults and the elderly. Since its validation, over 100 adults have performed the task. The MSIT produces a robust and temporally stable reaction time interference effect (range 200-350 ms), and single runs of the MSIT have produced CFP network activation in approximately 95% of tested subjects. The robust, reliable and temporally stable neuroimaging and performance data make the MSIT a useful task with which to study normal human cognition and psychiatric pathophysiology. PMID- 17406251 TI - Conjugation of chelating agents to proteins and radiolabeling with trivalent metallic isotopes. AB - Peptides and proteins may be tagged with metallic elements in order to use them as imaging reporters or for other applications. The polypeptide of interest is first conjugated to a suitable chelating agent that forms stable complexes with the element of interest. This conjugation step is undertaken either in aqueous or in non-aqueous conditions depending on the solubility of the substrate. For polypeptides of greater than approximately 10 kDa in size, this is normally done in aqueous medium. Most commonly the chelators are reacted with lysine amino groups. The protein is first desalted into a suitable buffer at pH 8-9 and a molar excess of a bifunctional chelating agent is added. After a suitable period of incubation, excess, unreacted or hydrolyzed chelator is removed and the protein conjugate is desalted into an acidic buffer. The conjugate can then be tagged by addition of a suitable metal salt followed, if necessary, by removal of unchelated metal. As described in the protocol that follows, the entire conjugation, purification and labeling procedure takes about 2 d. PMID- 17406252 TI - A method for C-terminal sequence analysis in the proteomic era (proteins cleaved with cyanogen bromide). AB - There is growing interest in the overall study of post-translational modifications (PTMs) of proteins. Beside phosphorylation and glycosylation, truncations of the nascent polypeptide chain at the N or C termini are by far the most common types of PTMs found in proteins. However, little attention has been paid to the development of approaches that allow a systematic analysis of these proteolytic processing events. Here we present a protocol that allows the identification of the C-terminal sequences of proteins. A peptide mixture is generated by cleavage of the protein with cyanogen bromide and is incubated with carboxypeptidase Y. The enzyme is only able to act on the C-terminal fragment, because this is the only peptide without a homoserine lactone residue at its C terminus. The resulting fragments, forming a peptide ladder, are analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The entire protocol, including the CNBr cleavage, takes 21 h and can be applied to proteins purified either by SDS-PAGE or by 2D PAGE or in solution. PMID- 17406253 TI - The bio-barcode assay for the detection of protein and nucleic acid targets using DTT-induced ligand exchange. AB - The recently developed bio-barcode assay for the detection of nucleic acid and protein targets without PCR has been shown to be extraordinarily sensitive, showing high sensitivity for both nucleic acid and protein targets. Two types of particles are used in the assay: (i) a magnetic microparticle with recognition elements for the target of interest; and (ii) a gold nanoparticle (Au-NP) with a second recognition agent (which can form a sandwich around the target in conjunction with the magnetic particle) and hundreds of thiolated single-strand oligonucleotide barcodes. After reaction with the analyte, a magnetic field is used to localize and collect the sandwich structures, and a DTT solution at elevated temperature is used to release the barcode strands. The barcode strands can be identified on a microarray via scanometric detection or in situ if the barcodes carry with them a detectable marker. The recent modification to the original bio-barcode assay method, utilizing DTT, has streamlined and simplified probe preparation and greatly enhanced the quantitative capabilities of the assay. Here we report the detailed methods for performing the ligand exchange bio barcode assay for both nucleic acid and protein detection. In total, reagent synthesis, probe preparation and detection require 4 d. PMID- 17406254 TI - Bioluminescence resonance energy transfer (BRET) for the real-time detection of protein-protein interactions. AB - A substantial range of protein-protein interactions can be readily monitored in real time using bioluminescence resonance energy transfer (BRET). The procedure involves heterologous coexpression of fusion proteins, which link proteins of interest to a bioluminescent donor enzyme or acceptor fluorophore. Energy transfer between these proteins is then detected. This protocol encompasses BRET1, BRET2 and the recently described eBRET, including selection of the donor, acceptor and substrate combination, fusion construct generation and validation, cell culture, fluorescence and luminescence detection, BRET detection and data analysis. The protocol is particularly suited to studying protein-protein interactions in live cells (adherent or in suspension), but cell extracts and purified proteins can also be used. Furthermore, although the procedure is illustrated with references to mammalian cell culture conditions, this protocol can be readily used for bacterial or plant studies. Once fusion proteins are generated and validated, the procedure typically takes 48-72 h depending on cell culture requirements. PMID- 17406255 TI - High-efficiency transfection and drug selection of genetically transformed blood stages of the rodent malaria parasite Plasmodium berghei. AB - This protocol describes a method of genetic transformation for the rodent malaria parasite Plasmodium berghei with a high transfection efficiency of 10(-3)-10(-4). It provides methods for: (i) in vitro cultivation and purification of the schizont stage;(ii) transfection of DNA constructs containing drug-selectable markers into schizonts using the nonviral Nucleofector technology; and (iii) injection of transfected parasites into mice and subsequent selection of mutants by drug treatment in vivo. Drug selection is described for two (antimalarial) drugs, pyrimethamine and WR92210. The drug-selectable markers currently in use are the pyrimethamine-resistant dihydrofolate reductase (dhfr) gene of Plasmodium or Toxoplasma gondii and the DHFR gene of humans that confer resistance to pyrimethamine and WR92210, respectively. This protocol enables the generation of transformed parasites within 10-15 d. Genetic modification of P. berghei is widely used to investigate gene function in Plasmodium, and this protocol for high-efficiency transformation will enable the application of large-scale functional genomics approaches. PMID- 17406256 TI - Physiological 3D tissue model of the airway wall and mucosa. AB - This protocol describes the setup, maintenance and characteristics of a tissue engineered model of the human bronchial mucosa that can be used for basic physiology and pathophysiology studies. The model includes a well-differentiated epithelium with functional cilia, mucus secretion and subepithelial fibroblasts within type I collagen. The tissue is created within porous polymeric wells to prevent gel contraction and allow culture at the air-liquid interface. It requires at least 2 wk to be established and can be maintained thereafter for over 4 wk, with tissue differentiation moving towards a more physiologically relevant phenotype with increasing time in culture. Over time, the extracellular matrix also remodels, depositing proteins such as types III and IV collagen and fibronectin. Because it recapitulates many key anatomical and functional features of the airway wall, this model is well suited for a wide range of studies, including those on airway remodeling, transepithelial transport and inflammatory cell interactions with the mucosa. The entire protocol takes 4-6 wk, including cell expansion, depending on the extent of ciliogenesis desired. PMID- 17406257 TI - In vitro measurement of cell death with the annexin A5 affinity assay. AB - One of the hallmarks of cell death is the cell surface-expression of phosphatidylserine. Expression of phosphatidylserine at the cell surface can be measured in vitro with the phosphatidylserine-binding protein annexin A5 conjugated to fluorochromes. This measurement can be made by flow cytometry or by confocal scanning-laser microscopy. The annexin A5 affinity assay comprises the incubation of cells stimulated to execute cell death with fluorescence-labeled annexin A5 and propidium iodide. Living cells are annexin A5-negative and propidium iodide negative, cells in the early phases of cell death are annexin A5 positive-and propidium iodide-negative, and secondary necrotic cells are annexin A5-positive and propidium iodide-positive. The entire procedure takes about 30 minutes for flow cytometry and 45 minutes for confocal scanning-laser microscopy. Various precautions and considerations are discussed further in the protocol described here. PMID- 17406258 TI - Locating potential enhancer elements by comparative genomics using the EEL software. AB - This protocol describes the use of Enhancer Element Locator (EEL), a computer program that was designed to locate distal enhancer elements in long mammalian sequences. EEL will predict the location and structure of conserved enhancers after being provided with two orthologous DNA sequences and binding specificity matrices for the transcription factors (TFs) that are expected to contribute to the function of the enhancers to be identified. The freely available EEL software can analyze two 1-Mb sequences with 100 TF motifs in about 15 min on a modern Windows, Linux or Mac computer. The output provides several hypotheses about enhancer location and structure for further evaluation by an expert on enhancer function. PMID- 17406259 TI - Determining protein kinase substrate specificity by parallel solution-phase assay of large numbers of peptide substrates. AB - We describe here a protocol for determining the activity of protein kinases on a large set of peptide substrates. Biotin-tagged peptides are arrayed in multiwell plates and incubated in solution with the kinase of interest and radiolabeled ATP. Reactions are then spotted simultaneously onto a streptavidin membrane, which is washed, dried, and analyzed by autoradiography or phosphor imaging. Differences in the extent of radiolabel incorporation into the various peptide substrates provide a measure of the sequence specificity of the kinase. This approach is a faster, more sensitive, and more generally applicable method for determining kinase phosphorylation motifs than older peptide library screening approaches based on Edman sequencing. The procedure is readily adaptable to other applications that require parallel processing of many kinase reactions, such as screening for small molecule inhibitors. In the format described here, preparation of stock plates prior to running the reactions will require about 4 days. Afterwards, the protocol takes approximately 6 hours to perform. PMID- 17406260 TI - Targeted bulk-loading of fluorescent indicators for two-photon brain imaging in vivo. AB - One of the challenges for modern neuroscience is to understand the rules of concerted neuronal function in vivo. This question can be addressed using noninvasive high-resolution imaging techniques like two-photon microscopy. This protocol describes a versatile approach for in vivo two-photon calcium imaging of neural networks, stained with membrane-permeant fluorescent-indicator dyes. It is based on a targeted pressure ejection of the dye into the tissue of interest and can be used for a large spectrum of indicator dyes, including Oregon Green 488 BAPTA-1 acetoxymethyl ester and Fura-2 acetoxymethyl ester. Through the use of dye mixtures and multicolor imaging, this technique allows the visualization of distinct neurons and glial cells up to 500 microm below the brain surface. It is suitable for staining the brain tissue of various different species (e.g., mouse, rat, cat and zebrafish) at all developmental stages. When combined with brain microendoscopy, it allows the monitoring of intracellular calcium signals in awake, behaving animals. The total time required to carry out the protocol, including dissection and cell staining, is approximately 2 h. Thereafter, imaging experiments might be performed for at least 6 h. PMID- 17406261 TI - Gas chromatography mass spectrometry-based metabolite profiling in plants. AB - The concept of metabolite profiling has been around for decades, but technical innovations are now enabling it to be carried out on a large scale with respect to the number of both metabolites measured and experiments carried out. Here we provide a detailed protocol for gas chromatography mass spectrometry (GC-MS) based metabolite profiling that offers a good balance of sensitivity and reliability, being considerably more sensitive than NMR and more robust than liquid chromatography-linked mass spectrometry. We summarize all steps from collecting plant material and sample handling to derivatization procedures, instrumentation settings and evaluating the resultant chromatograms. We also define the contribution of GC-MS-based metabolite profiling to the fields of diagnostics, gene annotation and systems biology. Using the protocol described here facilitates routine determination of the relative levels of 300-500 analytes of polar and nonpolar extracts in approximately 400 experimental samples per week per machine. PMID- 17406262 TI - Glycome mapping on DNA sequencing equipment. AB - Here we provide a detailed protocol for the analysis of protein-linked glycans on DNA sequencing equipment. This protocol satisfies the glyco-analytical needs of many projects and can form the basis of 'glycomics' studies, in which robustness, high throughput, high sensitivity and reliable quantification are of paramount importance. The protocol routinely resolves isobaric glycan stereoisomers, which is much more difficult by mass spectrometry (MS). Earlier methods made use of polyacrylamide gel-based sequencers, but we have now adapted the technique to multicapillary DNA sequencers, which represent the state of the art today. In addition, we have integrated an option for HPLC-based fractionation of highly anionic 8-amino-1,3,6-pyrenetrisulfonic acid (APTS)-labeled glycans before rapid capillary electrophoretic profiling. This option facilitates either two dimensional profiling of complex glycan mixtures and exoglycosidase sequencing, or MS analysis of particular compounds of interest rather than of the total pool of glycans in a sample. PMID- 17406263 TI - Generation of T-cell receptor retrogenic mice. AB - T-cell receptor (TCR) transgenic (Tg) mice have revolutionized our understanding of many aspects of T-cell biology. Whereas they provide an almost unlimited source of T cells with a single specificity, breeding them onto different backgrounds and/or new knockout/knock-in mouse models is often time-consuming (6 months to several years), which can make the process costly and can significantly delay research. This protocol describes a new method for expressing defined TCR alpha and TCR-beta proteins from a single 2A peptide-linked multicistronic retroviral vector in mice, using retrovirus-mediated stem cell gene transfer. We refer to these as 'retrogenic' (Rg) mice ('retro' from retrovirus and 'genic' from Tg) to avoid confusion with traditional transgenic mice. We have successfully used this approach to express over 50 different TCRs on several different mouse backgrounds in as little as 6 weeks. PMID- 17406264 TI - Blue native PAGE. AB - Blue native PAGE (BN-PAGE) can be used for one-step isolation of protein complexes from biological membranes and total cell and tissue homogenates. It can also be used to determine native protein masses and oligomeric states and to identify physiological protein-protein interactions. Native complexes are recovered from gels by electroelution or diffusion and are used for 2D crystallization and electron microscopy or analyzed by in-gel activity assays or by native electroblotting and immunodetection. In this protocol, we describe methodology to perform BN-PAGE followed by (i) native extraction or native electroblotting of separated proteins, or (ii) a second dimension of tricine-SDS PAGE or modified BN-PAGE, or (iii) a second dimension of isoelectric focusing (IEF) followed by a third dimension of tricine-SDS-PAGE for the separation of subunits of complexes. These protocols for 2D and 3D PAGE can be completed in 2 and 3 days. PMID- 17406265 TI - In vivo multimodal imaging of transplanted pancreatic islets. AB - Interest is increasing in the transplantation of pancreatic islets as a means to achieve insulin independence in individuals with type I diabetes. The success of this approach is hampered by the absence of methods to follow the fate of transplanted islets non-invasively. In vivo imaging seems to be the most appropriate technique to achieve this goal in small animals and eventually in humans. Here we describe a protocol for labeling and subsequent imaging of transplanted islets in vivo using magnetic resonance imaging (MRI) and optical imaging. The whole series of experiments can be carried out in roughly 48 h. We believe that our approach can significantly advance the current ability to determine islet distribution, and possibly survival, after transplantation. This information would be essential not only for the long-term monitoring of graft function but also for the design of improved transplantation and immunomodulatory methods. PMID- 17406266 TI - Silencing gene expression by targeting chromosomal DNA with antigene peptide nucleic acids and duplex RNAs. AB - The value of recognizing cellular RNA sequences by short interfering RNAs (siRNAs) in mammalian cells is widely appreciated, but what might be learned if it were also possible to recognize chromosomal DNA? Recognition of chromosomal DNA would have many applications, such as inhibiting gene expression, activating gene expression, introducing mutations, and probing chromosome structure and function. We have shown that antigene peptide nucleic acids (agPNAs) and antigene duplex RNAs (agRNAs) block gene expression and probe chromosomal DNA. Here we describe a protocol for designing antigene agents and introducing them into cells. This protocol can also be used to silence expression with PNAs or siRNAs that target mRNA. From preparation of oligomers to analysis of data, experiments with agPNAs and agRNAs require approximately 14 d and 9 d, respectively. PMID- 17406267 TI - Normal human keratinocyte assay to predict the human irritancy response. AB - The potential to cause human skin irritation responses is part of the required information for the registration of new chemicals and for chemicals to conform to the new EU Chemical regulations under Registration, Evaluation, and Authorisation of Chemicals (REACH). The Fund for Replacement of Animals in Medical Experiments (FRAME) has developed an in vitro assay for potential human irritant responses via the skin, employing human passage 1-3 primary keratinocytes. The assay is based on two assessments: a direct assay of keratinocytes and an assay of the effect on macrophages of the mediators released by the keratinocytes. Keratinocyte activity is measured by the resazurin assay. Mediator activity is measured by assaying the release of nitric oxide (NO) following macrophage stimulation. It is also possible to measure release of interleukin-1alpha (IL 1alpha). Additional mediators are also released, but the precise range and composition of all these mediators are not clear at present. From time of exposure, this assay can be completed in 3-7 d; the exact timing is dependent on the rate and degree of recovery. PMID- 17406268 TI - The 'reverse capture' autoantibody microarray: a native antigen-based platform for autoantibody profiling. AB - We have previously reported the development and the use of a 'reverse capture' autoantibody microarray for studies of antigen-autoantibody profiling. We developed the 'reverse capture' autoantibody microarray to allow the user to characterize and to compare autoantibody profiles. Based on the dual-antibody sandwich immunoassay of ELISA, our 'reverse capture' protocol facilitates the detection of autoimmunity to native host antigens. Our method has the advantage over traditional protein arrays of being able to detect autoimmunity to epitopes found on the post-translational modifications (PTMs) of native antigens. The first step of this method is to immobilize native antigens onto the monoclonal antibodies spotted on the array surface. Using the antigens captured by the microarray as 'baits,' we then incubate the array with differentially labeled IgG from test and control samples, and perform a two-slide dye-swap to normalize for dye effects. In this protocol we present a detailed description of the 'reverse capture' autoantibody microarray, a method that can be completed in 9-10 h over 1 2 d. PMID- 17406269 TI - Photoconductive stimulation of neurons cultured on silicon wafers. AB - Photoconductive stimulation allows the noninvasive depolarization of neurons cultured on a silicon wafer. This technique relies on a beam of light to target a cell of interest while applying a voltage bias across the silicon wafer. The targeted cell is excited with minimal physiological manipulation, and, therefore, long-term modulation of activity patterns and investigations of biochemical mechanisms sensitive to physiological perturbations are possible. Ideologically similar to transistor-based neuronal interfaces, the photoconductive-stimulation method has the advantage of being able to extracellularly excite any neuron in a network regardless of its spatial position on the silicon substrate. This protocol can be easily implemented on a conventional reflected-light fluorescence microscope using materials and resources that are readily available. Time requirements are comparable to standard cell-culture and electrophysiology techniques. When combined with fluorescence imaging of various molecular probes, activity-dependent cellular processes can be dynamically monitored. PMID- 17406270 TI - Real-time in vivo imaging of transgenic bioluminescent blood stages of rodent malaria parasites in mice. AB - This protocol describes a methodology for imaging the sequestration of infected erythrocytes of the rodent malaria parasite Plasmodium berghei in the bodies of live mice or in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of infected erythrocytes is performed by measuring bioluminescence produced by the enzymatic reaction between luciferase and its substrate luciferin, which is injected into the mice several minutes prior to imaging. The bioluminescence signal is detected by an intensified charge-coupled device (I-CCD) photon-counting video camera. Sequestration of infected erythrocytes is imaged during short-term infections with synchronous parasite development or during ongoing infections. With this technology, sequestration patterns of the schizont stage can be quantitatively analyzed within 1-2 d after infection. Real-time in vivo imaging of infected erythrocytes will provide increased insights into the dynamics of sequestration and its role in pathology, and can be used to evaluate strategies that prevent sequestration. PMID- 17406271 TI - A method for parental RNA interference in the wasp Nasonia vitripennis. AB - The wasp Nasonia vitripennis is emerging as a useful model organism in which to address a variety of biological questions, due, in part, to its ease of laboratory use, unique aspects of its biology and the sequencing of its genome. In order to take full advantage of the potential of this organism, methods for manipulating gene function are needed. To this end, a protocol for parental RNA interference (pRNAi) in N. vitripennis is described. pRNAi entails injecting pupae with double-stranded RNA, allowing the injected wasps to eclose and examining the progeny for developmental defects. This basic protocol is described in the context of the life cycle of N. vitripennis. This technique has been useful in elucidating the function of most, although not all, genes tested to date, and has potential applications beyond embryonic patterning. pRNAi experiments in Nasonia can be completed in as little as 2 weeks. PMID- 17406272 TI - The ecology of childhood overweight: a 12-year longitudinal analysis. AB - OBJECTIVE: To investigate ecological correlates of the development of overweight in a multisite study sample of children followed from age 2 to 12. DESIGN: Longitudinal examination of covariates of overweight status throughout childhood, with covariates drawn from three ecological levels: sociocultural or demographic, quality of the child's home environment, and proximal child experience that could directly affect the balance between energy intake and energy expenditure. SUBJECTS: A total of 960 children participating in a long-term longitudinal study provided growth data at least once; 653 of the children had complete data on covariates. MEASUREMENTS: Height and weight measured seven times between ages 2 and 12 were converted to a body mass index (BMI) and entered into a latent transition analysis to identify patterns of overweight across childhood. Ecological correlates measured longitudinally included demographic characteristics obtained by maternal report, home environment quality obtained by observation and maternal report, and proximal child experience factors obtained by observation, maternal report and child report. RESULTS: Four patterns of overweight were found: never overweight, overweight beginning at preschool age, overweight beginning in elementary school, and return to normal weight after being overweight at preschool age. The weight status groups differed on home environment quality and proximal child experience factors but not on demographics. Children overweight at preschool had less sensitive mothers than never overweight children. Children overweight at school age had fewer opportunities for productive activity at home than did never overweight children. School-age overweight children also watched the most TV after school. Multivariate logistic regression analyses further indicated the significance to children's weight status of proximal child experience variables. Less physically active children and those who watched more television after school were more likely to become overweight. Results did not vary by child sex. CONCLUSION: The results support the idea that childhood overweight is multiply determined. The one potentially important and changeable factor identified as a target for intervention centers on how children spend their time, especially their after school time. Children who are more physically active and spend less time watching TV after school are less likely to become overweight by age 12. PMID- 17406273 TI - Developmental changes in adiposity in toddlers and preschoolers in the GENESIS study and associations with the ACE I/D polymorphism. AB - OBJECTIVES: To investigate the relationship between the angiotensin I-converting enzyme 1 (ACE) I/D polymorphism and adiposity-related phenotypes in a large cohort of toddlers and preschoolers. METHODS: Body composition measurements and DNA samples were obtained from 2102 Greek children aged 1-6 years, as part of a large-scale epidemiological study (GENESIS). All children were genotyped for the ACE I/D polymorphism and gender- and age-stratified statistical analyses were performed. RESULTS: In girls aged 4-6 years, the D-allele was associated with higher measurements of body mass index (BMI) (P=0.018), waist (P=0.001) and upper arm (P=0.013) circumferences, genotype accounting for 2.5, 4 and 3% of the phenotypic variance, respectively. In boys, the D-allele showed strong associations with lower BMI (P=0.001) at the age of 1-2 years that explained 17% of the phenotypic variance and with larger suprailiac skinfold (P=0.008) at 3-4 years old that explained 2% of the variance. No other significant associations between the ACE I/D polymorphism and adiposity-related phenotypes were found. In girls, the age at which significant associations were revealed coincided with the age at which BMI was observed to increase after its developmental nadir, but this feature of the association was not observed in boys. CONCLUSIONS: The ACE I/D polymorphism is associated with developmental and physiological changes in adiposity-related traits during early childhood in a gender- and age-specific manner. PMID- 17406275 TI - Differentiation of mouse embryonic stem cells to insulin-producing cells. AB - Here, we describe a basic protocol for the in vitro differentiation of mouse embryonic stem (ES) cells into insulin-producing cells. The three-step protocol comprises (i) the formation of embryoid bodies, (ii) the spontaneous differentiation of embryoid bodies into progenitor cells of ecto-, meso- and endodermal lineages, and (iii) the induction of differentiation of early progenitors into the pancreatic lineage. Differentiated cells can be obtained within approximately 33 d. Differentiation induction by growth and extracellular matrix factors, including laminin, nicotinamide and insulin, leads to the formation of ES-derived progeny that resembles cells committed to the pancreatic lineage. During differentiation, transcript levels of genes expressed in early pancreatic cells are upregulated. Continued differentiation results in the development of C-peptide/insulin-positive islet-like clusters that release insulin upon glucose stimulation. Differentiated ES cells that overexpress the pancreatic developmental control gene Pax4 develop insulin-secretory granules and reveal functional properties with respect to the pancreas-specific ATP-modulated K+ channel and the normalization of glycemia of streptozotocin-treated diabetic mice. PMID- 17406276 TI - Rational design and in vitro and in vivo delivery of Dicer substrate siRNA. AB - RNA interference is a powerful tool for target-specific knockdown of gene expression. The triggers for this process are duplex small interfering RNAs (siRNAs) of 21-25 nt with 2-bp 3' overhangs produced in cells by the RNase III family member Dicer. We have observed that short RNAs that are long enough to serve as Dicer substrates (D-siRNA) can often evoke more potent RNA interference than the corresponding 21-nt siRNAs; this is probably a consequence of the physical handoff of the Dicer-produced siRNAs to the RNA-induced silencing complex. Here we describe the design parameters for D-siRNAs and a protocol for in vitro and in vivo intraperitoneal delivery of D-siRNAs and siRNAs to macrophages. siRNA delivery and transfection and analysis of macrophages in vivo can be accomplished within 36 h. PMID- 17406277 TI - Southern blotting. AB - This protocol describes a basic method to perform the Southern blot. Blotting allows the detection of specific molecules among a mixture separated by gel electrophoresis. Molecules are transferred from the gel to a porous membrane by capillary action using absorbent paper to soak solution through the gel and the membrane. For DNA, specific sequences are detected in the membrane by molecular hybridization with labeled nucleic acid probes. The original method, on which this protocol is based, used labeled RNAs to detect specific DNA fragments in genomic DNA that had been digested with restriction endonucleases. This protocol can be completed in 1-5 d and is inexpensive to carry out, as it requires only basic laboratory equipment. PMID- 17406278 TI - Localized transfection with magnetic beads coated with PCR products and other nucleic acids. AB - The bead transfection method involves binding nucleic acids onto 3-microm diameter paramagnetic beads, treating the beads with transfection reagent, and using them as scaffolds to direct transfection to individual cells or regions in a population. Typically, PCR products are used because they can be conveniently generated using biotinylated primers and can introduce site-directed mutations, without the need for cloning or plasmid purification. However, the method can be adapted to transfect plasmid DNA or RNA. The magnetic properties of the beads allows magnets to direct the loci of transfection in cell culture; magnetic arrays are built in cell culture chambers to allow multiple parallel transfections on the same microscope coverslip. The PCR reaction and transfection can be carried out in 1 d, and transfection results can be viewed in 24-48 h. PMID- 17406279 TI - Blind patch clamp recordings in embryonic and adult mammalian brain slices. AB - To obtain electrophysiological recordings in brain slices, sophisticated and expensive pieces of equipment can be used. However, costly microscope equipment with infrared differential interference contrast optics is not always necessary or even desirable. For instance, obtaining a randomized unbiased sample in a given preparation would be better accomplished if cells were not directly visualized before recording. In addition, some preparations require thick slices, and direct visualization is not possible. Here we describe a protocol for the 'blind patch clamp method' that we developed several years ago to perform electrophysiological recordings in mammalian brain slices using a standard patch clamp amplifier, dissecting microscope and recording chamber. Overall, it takes approximately 3-4 h to set up this procedure. PMID- 17406280 TI - A lectin microarray approach for the rapid analysis of bacterial glycans. AB - Rapid evaluation of microbial cell-surface carbohydrates is essential to understanding the mechanisms by which bacteria use glycans to establish pathogenic or symbiotic relationships. Microbial glycan analysis is complicated both by the vast diversity of possible carbohydrate structures and by their dynamic nature. Bacteria can rapidly alter their glycan coats by switching the genes that are involved on and off in a phase-variable manner. Currently, there is a lack of appropriate tools for studying dynamic carbohydrate alterations. Here, we present a lectin microarray protocol for the high-throughput evaluation of cell-surface microbial sugars. The binding patterns of fluorescent bacteria to these arrays provide a simple means to fingerprint bacteria based on their surface carbohydrates. In addition, this method provides a rapid, parallel evaluation of glycans from multiple bacterial samples, allowing dynamic changes in carbohydrate structures to be studied. The entire procedure takes approximately 12 h but the printing of the microarray can be performed in advance. PMID- 17406281 TI - An autonomous fueled machine that replicates catalytic nucleic acid templates for the amplified optical analysis of DNA. AB - Here we describe a protocol for the amplified detection of a target DNA using a DNA/FokI-based replicating cutting machine. The protocol is based on the design of a sensing hairpin oligonucleotide that is opened upon hybridization with the analyte DNA. The endonuclease FokI binds to the double-stranded complex and cleaves it to a "cutter" unit. The "cutter" unit reacts with a fuel oligonucleotide to generate and amplify the signal. The fuel molecule is an oligonucleotide in a hairpin configuration with a fluorophore/quencher pair attached to the 5' and 3' ends. Formation of the duplex between the cutter and the fuel leads to the scission of the duplex by FokI, leading to a second, replicated "cutter", a fluorescent waste product, and to the regeneration of the original "cutter" unit. The autonomous replication of the "cutter" unit, as a result of the primary recognition of the analyte DNA, leads to the amplified fluorescent detection of the analyte DNA with a sensitivity limit of 1 x 10(-14) M. The operation of the machine and the sensing process are monitored by the fluorescence generated by the waste product. Here we apply the protocol, which takes about 2 h to complete, to analyze a Tay-Sachs genetic disorder mutant DNA. PMID- 17406282 TI - Analysis of locomotor activity rhythms in Drosophila. AB - The genetic, molecular and anatomical dissection of the circadian clock in Drosophila and other higher organisms relies on the quantification of rhythmic phenotypes. Here, we introduce the methods currently in use in our laboratories for the analysis of fly locomotor activity rhythms. This phenotype provides a relatively simple, automated, efficient, reliable and robust output for the circadian clock. Thus it is not surprising that it is the preferred readout for measuring rhythmicity under a variety of conditions for most fly clock laboratories. The procedure requires at least 10 days of data collection and several days for analysis. In this protocol we advise on fly maintenance and on experimental design when studying the genetics of behavioral traits. We describe the setup for studying locomotor activity rhythms in the fruit fly and we introduce the statistical methods in use in our laboratories for the analysis of periodic data. PMID- 17406283 TI - A microarray-based protocol for monitoring the growth of yeast overexpression strains. AB - Gene overexpression can be used to investigate the biological pathways that are important in the response to a small molecule or other perturbation. To facilitate the use of gene overexpression in the study of small-molecule mechanisms, we developed a microarray-based protocol for monitoring the growth of a pool of yeast strains, each overexpressing a different protein. In this protocol, yeast harboring a set of approximately 3,900 galactose-inducible overexpression plasmids are grown in the absence or presence of a small molecule for multiple generations. The plasmids are then extracted from the two populations, processed and labeled in such a manner that their relative concentrations can be determined by competitive hybridization to a microarray. Although this protocol was developed for monitoring a specific set of overexpression plasmids, it could presumably be adapted to monitor yeast that have been transformed with any set of plasmids for which the gene inserts have been spotted, or otherwise arrayed, in a microarray format. This protocol can be completed in approximately 15 hours of hands-on time over the course of several days. PMID- 17406284 TI - Immunoprecipitation of mRNA-protein complexes. AB - Immunoprecipitation of mRNA-protein complexes is a method that can be used to study RNA binding protein (RBP)-RNA interactions. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. It can also be used as a second, independent method to verify RBP-mRNA interactions discovered through more universal screening techniques. We describe the immunoprecipitation protocol in practical detail and discuss variations of the method as well as issues associated with it. The procedure takes three days to complete. PMID- 17406285 TI - The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol chloroform extraction: twenty-something years on. AB - Since its introduction, the 'single-step' method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years. PMID- 17406286 TI - Laser-capture microdissection. AB - Deciphering the cellular and molecular interactions that drive disease within the tissue microenvironment holds promise for discovering drug targets of the future. In order to recapitulate the in vivo interactions thorough molecular analysis, one must be able to analyze specific cell populations within the context of their heterogeneous tissue microecology. Laser-capture microdissection (LCM) is a method to procure subpopulations of tissue cells under direct microscopic visualization. LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, RNA transcript profiling, cDNA library generation, proteomics discovery and signal-pathway profiling. Herein we provide a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1-1.5 h. PMID- 17406287 TI - Tagging recombinant proteins with a Sel-tag for purification, labeling with electrophilic compounds or radiolabeling with 11C. AB - Selenocysteine (Sec; U in one-letter code) is the twenty-first naturally occurring amino acid, with a selenium atom that gives this cysteine (Cys) homolog unique biochemical properties, including a high nucleophilicity and significant reactivity with electrophilic agents. This can be used in biotechnological Sec dependent applications. Here, we describe how Sec can be introduced into a carboxy-terminal tetrapeptide motif (-Gly-Cys-Sec-Gly-COOH, known as a Sel-tag) for recombinant proteins by tailoring the encoding gene to become compatible with the Escherichia coli selenoprotein synthesis machinery. We also describe how the Sel-tag can be used as a basis for efficient one-step protein purification, rapid Sec-targeting protein labeling with electrophilic compounds, or radiolabeling with the positron emitter 11C. PMID- 17406288 TI - Selection by flow-sorting of genetically transformed, GFP-expressing blood stages of the rodent malaria parasite, Plasmodium berghei. AB - This protocol describes a methodology for the genetic transformation of the rodent malaria parasite Plasmodium berghei and the subsequent selection of transformed parasites expressing green fluorescent protein (GFP) by flow-sorting. It provides methods for: transfection of the schizont stage with DNA constructs that contain gfp as the selectable marker; selection of fluorescent mutants by flow-sorting; and injection of flow-sorted, GFP-expressing parasites into mice and the subsequent collection of transformed parasites. The use of two different promoters for the expression of GFP is described; these two promoters require slightly different procedures for the selection of mutants. The protocol enables the collection of transformed parasites within 10-12 days after transfection. The genetic modification of P. berghei is widely used to investigate gene function in Plasmodium sp. The application of flow-sorting to the selection of transformed parasites increases the possibilities of parasite mutagenesis, by effectively expanding the range of selectable markers. PMID- 17406289 TI - High-throughput plasmid cDNA library screening. AB - Libraries of cDNA clones are valuable resources for analyzing the expression, structure and regulation of genes, and for studying protein functions and interactions. Full-length cDNA clones provide information about intron and exon structures, splice junctions, and 5' and 3' untranslated regions (UTRs). Open reading frames (ORFs) derived from cDNA clones can be used to generate constructs allowing the expression of both wild-type proteins and proteins tagged at their amino or carboxy terminus. Thus, obtaining full-length cDNA clones and sequences for most or all genes in an organism is essential for understanding genome functions. EST sequencing samples cDNA libraries at random, an approach that is most useful at the beginning of large-scale screening projects. As projects progress towards completion, however, the probability of identifying unique cDNAs by EST sequencing diminishes, resulting in poor recovery of rare transcripts. Here we describe an adapted, high-throughput protocol intended for the recovery of specific, full-length clones from plasmid cDNA libraries in 5 d. PMID- 17406290 TI - Enhanced delivery of cell-penetrating peptide-peptide nucleic acid conjugates by endosomal disruption. AB - Improvement of cellular uptake and cellular localization is still one of the main obstacles to the development of antisense-antigene therapeutics, including peptide nucleic acid (PNA). Cell-penetrating peptides (CPPs) such as Tat peptide and polyarginine have been widely used to improve the cellular uptake of PNA and other antisense agents. Cellular uptake of most CPP conjugates occurs mainly through endocytotic pathways, and most CPP conjugate is retained in the endosomal compartments of the cell. Several methods to induce endosome disruption have been shown to improve the bioavailability of CPP conjugates to the cytosol and/or nucleus by facilitating escape from the endosomal compartments. Here we describe protocols for the delivery of CPP-PNA conjugates to adherent cultured cells using photodynamic treatment (photochemical internalization), Ca2+ treatment or chloroquine treatment to potentiate the antisense effects of CPP-PNA conjugates through increased release of CPP conjugates into the cytoplasm. This protocol, consisting of CPP-mediated delivery assisted by an endosome-disruption agent, allows the delivery of the CPP-PNA conjugates to the nucleus and/or cytosol of cultured cells. The endosome-disruption treatment improves the nuclear antisense effects of CPP-PNA conjugates by up to two orders of magnitude using 24-hour delivery. PMID- 17406291 TI - Isolation of specific RNA-binding proteins using the streptomycin-binding RNA aptamer. AB - Here we report a simple and cheap one-step affinity purification protocol for isolating RNAs or proteins that interact with selected functional RNAs. The streptomycin-binding aptamer, termed 'StreptoTag,' is embedded in or fused to either end of any RNA of interest. The resulting hybrid RNA can then be immobilized on a streptomycin affinity matrix. When a complex protein mixture or total cellular lysate is applied to the matrix, subsequent elution with free streptomycin allows efficient recovery of specific ribonucleoprotein or RNA-RNA complexes. The method was successfully used to purify yeast and phage RNA-binding proteins and group II intron, viral and bacterial noncoding RNA (ncRNA)-binding proteins. The selective enrichment of bacterial mRNAs that bind ncRNAs has also been demonstrated. Once the affinity matrix, the RNA construct and the protein extracts have been prepared, the experimental procedure can be performed in 1-2 h. PMID- 17406292 TI - Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method. AB - Collective efforts of several laboratories in the past two decades have resulted in the development of various methods for Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana. Among these, the floral dip method is the most facile protocol and widely used for producing transgenic Arabidopsis plants. In this method, transformation of female gametes is accomplished by simply dipping developing Arabidopsis inflorescences for a few seconds into a 5% sucrose solution containing 0.01-0.05% (vol/vol) Silwet L-77 and resuspended Agrobacterium cells carrying the genes to be transferred. Treated plants are allowed to set seed which are then plated on a selective medium to screen for transformants. A transformation frequency of at least 1% can be routinely obtained and a minimum of several hundred independent transgenic lines generated from just two pots of infiltrated plants (20-30 plants per pot) within 2-3 months. Here, we describe the protocol routinely used in our laboratory for the floral dip method for Arabidopsis transformation. Transgenic Arabidopsis plants can be obtained in approximately 3 months. PMID- 17406293 TI - Two-photon targeted patching (TPTP) in vivo. AB - Two-photon-excited fluorescence laser-scanning microscopy (2PLSM) has provided a wealth of information about the spatiotemporal properties of biological processes at the single cell and population level. Because such nonlinear optical methods allow for imaging deep within biological tissue, 2PLSM can be combined with patch clamp techniques to obtain electrophysiological recordings from specific fluorescently labeled cells in vivo. Here a protocol referred to as two-photon targeted patching (TPTP) describes a method that may be used to record from cells in the intact animal labeled by virtually any type of fluorophore. We target neurons that have been optically and genetically identified using green fluorescent protein (GFP) expressed under the control of a specific promoter. TPTP when combined with genetic approaches therefore permits electrophysiological recordings from specified neurons and their compartments, including dendrites. This technique may be repeated in the same preparation many times over the course of several hours and is equally applicable to non-neuronal cell types. PMID- 17406294 TI - Reverse engineering cellular networks. AB - We describe a computational protocol for the ARACNE algorithm, an information theoretic method for identifying transcriptional interactions between gene products using microarray expression profile data. Similar to other algorithms, ARACNE predicts potential functional associations among genes, or novel functions for uncharacterized genes, by identifying statistical dependencies between gene products. However, based on biochemical validation, literature searches and DNA binding site enrichment analysis, ARACNE has also proven effective in identifying bona fide transcriptional targets, even in complex mammalian networks. Thus we envision that predictions made by ARACNE, especially when supplemented with prior knowledge or additional data sources, can provide appropriate hypotheses for the further investigation of cellular networks. While the examples in this protocol use only gene expression profile data, the algorithm's theoretical basis readily extends to a variety of other high-throughput measurements, such as pathway specific or genome-wide proteomics, microRNA and metabolomics data. As these data become readily available, we expect that ARACNE might prove increasingly useful in elucidating the underlying interaction models. For a microarray data set containing approximately 10,000 probes, reconstructing the network around a single probe completes in several minutes using a desktop computer with a Pentium 4 processor. Reconstructing a genome-wide network generally requires a computational cluster, especially if the recommended bootstrapping procedure is used. PMID- 17406295 TI - The JAM Test and its daughter P-JAM: simple tests of DNA fragmentation to measure cell death and stasis. AB - Cytotoxic T lymphocytes, and other death-inducing agents, have at least two different ways of killing their targets: drilling holes in the target cell membrane, or triggering the targets to commit suicide. The JAM Test is a method that measures the DNA fragmentation that accompanies cell suicide. We label target cells with radioactive DNA-precursor nucleotides and harvest them onto fiberglass filters, which trap large pieces of DNA but pass smaller fragments of apoptotic cells. As a general measure of apoptosis, the JAM Test described here is faster (can be completed in 4 h [or less if labeling is done the night before]), more quantitative, easier, more sensitive, more flexible and cheaper than most other current assays of apoptosis. The P-JAM, also discussed, additionally allows for assessment of death in cells that don't fragment their DNA, and allows for assays of agents that induce cell stasis rather than death. PMID- 17406296 TI - NMR: prediction of protein flexibility. AB - We present a protocol for predicting protein flexibility from NMR chemical shifts. The protocol consists of (i) ensuring that the chemical shift assignments are correctly referenced or, if not, performing a reference correction using information derived from the chemical shift index, (ii) calculating the random coil index (RCI), and (iii) predicting the expected root mean square fluctuations (RMSFs) and order parameters (S2) of the protein from the RCI. The key advantages of this protocol over existing methods for studying protein dynamics are that (i) it does not require prior knowledge of a protein's tertiary structure, (ii) it is not sensitive to the protein's overall tumbling and (iii) it does not require additional NMR measurements beyond the standard experiments for backbone assignments. When chemical shift assignments are available, protein flexibility parameters, such as S2 and RMSF, can be calculated within 1-2 h using a spreadsheet program. PMID- 17406297 TI - Identification and detection of RNA-RNA interactions using the yeast RNA hybrid system. AB - To test RNA-RNA interactions in cells, we developed a yeast RNA hybrid system derived from the yeast three-hybrid system. In this setup, the activation of a reporter gene (HIS3 or lacZ) is dependent on the interaction of two RNAs. One ('RNA X') is fused to MS2 RNA, forming the bait, which binds to a fusion protein composed of the MS2 coat and the LexA proteins. The second ('RNA Y') is fused to an RNA-based transcriptional activator (m26-11), forming the prey. If prey (RNA Y) binds to bait (RNA X), the m26-11 transcriptional activator is tethered to the promoter of the reporter genes. This protocol describes how to use this RNA hybrid system. In addition to testing RNA-RNA interactions, it can also be used to screen RNA libraries to identify new interaction partners or for mutational analysis of two known interaction partners. PMID- 17406298 TI - High Ca2+-phosphate transfection efficiency in low-density neuronal cultures. AB - This protocol describes a high-efficiency Ca2+-phosphate transfection method with low cell toxicity. The Ca2+-phosphate transfection method is widely used in transfecting neurons because of its low cell toxicity and simplicity in use, but the efficiency is typically low (approximately 1-5%). To solve this problem we have developed a new Ca2+-phosphate transfection protocol that increases the efficiency by 10-fold (< or = 60%), while maintaining low cell toxicity. First, it is critical to have gentle mixing of the DNA-Ca2+ solution with phosphate buffer to form a homogeneous snowlike precipitate (particle size 1-3 microm). Second, the precipitate should be dissolved using a slightly acidic culture medium to reduce cell toxicity. The high efficiency of this new protocol makes it possible to transfect single autaptic neurons as well as mature neurons (15-82 days in vitro) for gene functional analysis. The total time required for the protocol is 2-4 h (including 45 min-3 h incubation time). PMID- 17406299 TI - Construction of diverse adeno-associated viral libraries for directed evolution of enhanced gene delivery vehicles. AB - Rational design of improved gene delivery vehicles is a challenging and potentially time-consuming process. As an alternative approach, directed evolution can provide a rapid and efficient means for identifying novel proteins with improved function. Here we describe a methodology for generating very large, random adeno-associated viral (AAV) libraries that can be selected for a desired function. First, the AAV2 cap gene is amplified in an error-prone PCR reaction and further diversified through a staggered extension process. The resulting PCR product is then cloned into pSub2 to generate a diverse (>10(6)) AAV2 plasmid library. Finally, the AAV2 plasmid library is used to package a diverse pool of mutant AAV2 virions, such that particles are composed of a mutant AAV genome surrounded by the capsid proteins encoded in that genome, which can be used for functional screening and evolution. This procedure can be performed in approximately 2 weeks. PMID- 17406300 TI - Preparation of N-succinimidyl 3-[*I]iodobenzoate: an agent for the indirect radioiodination of proteins. AB - A procedure for the synthesis of N-succinimidyl 3-iodobenzoate labeled with any iodine isotope ([*I]SIB), which is an agent used in the radioiodination of proteins and peptides, from its tin precursor N-succinimidyl 3-(tri-n butylstannyl)benzoate (STB) is described. Also included are protocols for the synthesis of an unlabeled standard of SIB and the tin precursor. Radioiododestannylation of STB using tert-butylhydroperoxide as the oxidant gives [*I]SIB in 80% radiochemical yields. The total time for the synthesis of [*I]SIB from STB is approximately 95 min. Use of [*I]SIB yields radioiodinated proteins that are considerably more stable in vivo than those radioiodinated by the direct electrophilic method. PMID- 17406301 TI - Profluorescent protein fragments for fast bimolecular fluorescence complementation in vitro. AB - Here, we present a protocol for isolating the large N-terminal fragment of enhanced green fluorescent protein (EGFP) with a preformed chromophore. By itself, the chromophore-containing EGFP fragment exhibits very weak fluorescence, but it rapidly becomes brightly fluorescent upon complementation with the corresponding small, C-terminal EGFP fragment. Each EGFP fragment is cloned and overexpressed in E. coli as a fusion with self-splitting intein. After solubilizing and refolding these fusions from inclusion bodies, both EGFP fragments are cleaved from intein and purified using chitin columns. When these EGFP fragments are linked with the two complementary oligonucleotides and combined in equimolar amounts, fluorescence develops within a few minutes. The isolation of profluorescent protein fragments from recombinant E. coli cells requires approximately 3 d, and their conjugation to oligonucleotides requires 1 4 h. PMID- 17406302 TI - Multiplex amplification of ancient DNA. AB - This method is designed to assemble long, continuous DNA sequences using minimal amounts of fragmented ancient DNA as template. This is achieved by a two-step approach. In the first step, multiple fragments are simultaneously amplified in a single multiplex reaction. Subsequently, each of the generated fragments is amplified individually using a single primer pair, in a standard simplex (monoplex) PCR. The ability to amplify multiple fragments simultaneously in the first step allows the generation of large amounts of sequence from rare template DNA, whereas the second nested step increases specificity and decreases amplification of contaminating DNA. In contrast to current protocols using many template-consuming simplex PCRs, the method described allows amplification of several kilobases of sequence in just one reaction. It thus combines optimal template usage with a high specificity and can be performed within a day. PMID- 17406303 TI - Chromatin immunoprecipitation and microarray-based analysis of protein location. AB - Genome-wide location analysis, also known as ChIP-Chip, combines chromatin immunoprecipitation and DNA microarray analysis to identify protein-DNA interactions that occur in living cells. Protein-DNA interactions are captured in vivo by chemical crosslinking. Cell lysis, DNA fragmentation and immunoaffinity purification of the desired protein will co-purify DNA fragments that are associated with that protein. The enriched DNA population is then labeled, combined with a differentially labeled reference sample and applied to DNA microarrays to detect enriched signals. Various computational and bioinformatic approaches are then applied to normalize the enriched and reference channels, to connect signals to the portions of the genome that are represented on the DNA microarrays, to provide confidence metrics and to generate maps of protein-genome occupancy. Here, we describe the experimental protocols that we use from crosslinking of cells to hybridization of labeled material, together with insights into the aspects of these protocols that influence the results. These protocols require approximately 1 week to complete once sufficient numbers of cells have been obtained, and have been used to produce robust, high-quality ChIP chip results in many different cell and tissue types. PMID- 17406304 TI - Isotope labeling strategies for the study of high-molecular-weight proteins by solution NMR spectroscopy. AB - The development of isotope labeling methodology has had a significant impact on NMR studies of high-molecular-weight proteins and macromolecular complexes. Here we review some of this methodology that has been developed and used in our laboratory. In particular, experimental protocols are described for the production of highly deuterated, uniformly 15N- and 13C-labeled samples of large proteins, with optional incorporation of selective isotope labels into methyl groups of isoleucine, leucine and valine residues. Various types of methyl labeling schemes are assessed, and the utility of different methyl labeling strategies is highlighted for studies ranging from protein structure determination to the investigation of side-chain dynamics. In the case of malate synthase G (MSG), the time frame of the whole preparation, including the protein refolding step, is about 70 h. PMID- 17406305 TI - Isolating and engineering human antibodies using yeast surface display. AB - This protocol describes the process of isolating and engineering antibodies or proteins for increased affinity and stability using yeast surface display. Single chain antibody fragments (scFvs) are first isolated from an existing nonimmune human library displayed on the yeast surface using magnetic-activated cell sorting selection followed by selection using flow cytometry. This enriched population is then mutagenized, and successive rounds of random mutagenesis and flow cytometry selection are done to attain desired scFv properties through directed evolution. Labeling strategies for weakly binding scFvs are also described, as well as procedures for characterizing and 'titrating' scFv clones displayed on yeast. The ultimate result of following this protocol is a panel of scFvs with increased stability and affinity for an antigen of interest. PMID- 17406306 TI - Sample extraction techniques for enhanced proteomic analysis of plant tissues. AB - Major improvements in proteomic techniques in recent years have led to an increase in their application in all biological fields, including plant sciences. For all proteomic approaches, protein extraction and sample preparation are of utmost importance for optimal results; however, extraction of proteins from plant tissues represents a great challenge. Plant tissues usually contain relatively low amounts of proteins and high concentrations of proteases and compounds that potentially can limit tissue disintegration and interfere with subsequent protein separation and identification. An effective protein extraction protocol must also be adaptable to the great variation in the sets of secondary metabolites and potentially contaminating compounds that occurs between tissues (e.g., leaves, roots, fruit, seeds and stems) and between species. Here we present two basic protein extraction protocols that have successfully been used with diverse plant tissues, including recalcitrant tissues. The first method is based on phenol extraction coupled with ammonium acetate precipitation, and the second is based on trichloroacetic acid (TCA) precipitation. Both extraction protocols can be completed within 2 d. PMID- 17406307 TI - Subcellular imaging in the live mouse. AB - Fluorescent proteins are available in multiple colors and have properties such as intrinsic brightness and high quantum yield that make them optimally suited for in vivo imaging with subcellular resolution in the live mouse. In this protocol, cancer cells in live mice are labeled with green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in the cytoplasm. GFP nuclear labeling is effected by linkage of GFP to histone H2B, and a retroviral vector is used for cytoplasmic labeling with RFP. Double-labeled cells are injected by various methods. High-resolution imaging systems with microscopic optics, in combination with reversible skin flaps over various organs, enable the imaging of dual-color labeled cells at the subcellular level in live animals. The double transfection and selection procedures described here take 6-8 weeks. Cancer cell trafficking, deformation, extravasation, mitosis and cell death can be imaged with clarity. PMID- 17406308 TI - Cross-species chromosome painting. AB - Comparative genomics is an important and expanding field of research, and the genome-wide comparison of the chromosome constitution of different species makes a major contribution to this field. Cross-species chromosome painting is a powerful technique for establishing chromosome homology maps, defining the sites of chromosome fusions and fissions, investigating chromosome rearrangements during evolution and constructing ancestral karyotypes. Here the protocol for cross-species chromosome painting is presented. It includes sections on cell culture and metaphase preparation, labeling of chromosome-specific DNA, fluorescent in situ hybridization (chromosome painting) and image analysis. Cell culture and metaphase preparation can take between 1 and 2 wk depending on the cell culture. Labeling of chromosome-specific DNA is completed in 1 d. Fluorescent in situ hybridization can be completed in a maximum of 4 d. PMID- 17406309 TI - PCR-based accurate synthesis of long DNA sequences. AB - Here we describe a simple and rapid method for assembly and PCR-based accurate synthesis (PAS) of long DNA sequences. The PAS protocol involves the following five steps: (i) design of the DNA sequence to be synthesized and of 60-bp overlapping oligonucleotides to cover the entire DNA sequence; (ii) purification of the oligonucleotides by PAGE; (iii) first PCR, to synthesize DNA fragments of 400-500 bp in length using 10 inner (template) and two outer (primer) oligonucleotides; (iv) second PCR, to assemble the products of the first PCR into the full-length DNA sequence; and (v) cloning and verification of the synthetic DNA by sequencing and, if needed, error correction using an overlap-extension PCR technique. This method, which takes approximately 1 wk, is suitable for synthesizing diverse types of long DNA molecule. We have successfully synthesized DNA fragments from 0.5 to 12.0 kb, with high G+C content, repetitive sequences or complex secondary structures. The PAS protocol therefore provides a simple, rapid, reliable and relatively inexpensive method for synthesizing long, accurate DNA sequences. PMID- 17406310 TI - Labeling of aliphatic carboxylic acids using [11C]carbon monoxide. AB - Here we present a protocol for labeling aliphatic carboxylic acids with the positron-emitting radionuclide 11C (t(1/2) = 20.4 min) at the carboxyl position using [11C]carbon monoxide via photoinitiated free radical-mediated carbonylation. A solution of an alkyl iodide in a homogenous binary organic solvent-water mixture is introduced into a high-pressure photochemical reactor containing [11C]carbon monoxide. Then the reactor contents are pressurized to 40 MPa and irradiated with ultraviolet light for 6 min. The labeled product is purified using HPLC. All manipulations with radioactivity including the labeling synthesis are carried out on an automated Synthia system. In a typical case, 3.19 GBq of purified [1-(11)C]1,10-decanedicarboxylic acid (with a specific radioactivity of 188 GBq/micromol) can be obtained within 35 min after the end of a 10-microAh bombardment. Compared to previous labeling methods, this protocol is compatible with a wider range of functional groups, utilizes less-sensitive precursors, and is less subject to isotopic dilution. PMID- 17406311 TI - Generation of cells expressing improved doxycycline-regulated reverse transcriptional transactivator rtTA2S-M2. AB - Tet-on cell lines engineered to stably express doxycycline (Dox)-regulated reverse transcriptional transactivator (rtTA) have many applications in biomedical research and biotechnology. Unfortunately, construction and maintenance of such cells often proves to be costly, labor intensive and ineffective. Moreover, the Tet-on clones generated using standard methodology were often unstable and frequently displayed significantly changed physiological properties compared with their parental cells. Here we describe an optimized protocol for generation of Tet-on cells. The protocol is based on the use of a recently developed pN1p beta actin-rtTA2S-M2-IRES-EGFP vector (where IRES is an internal ribosome entry site) and permits relatively inexpensive construction of many Tet-on clones with essentially 100% efficiency. The method is well suited for 'difficult' cell lines displaying genetic instability and high levels of epigenetic silencing. The constructed Tet-on cells remain stable with time in the absence of any selection agents, are easy to monitor and preserve the characteristics of parental cells. The protocol can be completed in 2 months. PMID- 17406312 TI - State-of-the-art two-dimensional gel electrophoresis: a key tool of proteomics research. AB - Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) is the most popular and versatile method of protein separation among a rapidly growing array of proteomics technologies. Based on two distinct procedures, it combines isoelectric focusing (IEF), which separates proteins according to their isoelectric point (pI), and SDS-PAGE, which separates them further according to their molecular mass. At present, 2D-PAGE is capable of simultaneously detecting and quantifying up to several thousand protein spots in the same gel image. Here we provide comprehensive step-by-step instructions for the application of a standardized 2D-PAGE protocol to a sample of human plasma or cerebrospinal fluid (CSF). The method can be easily adapted to any type of sample. This four-day protocol provides detailed information on how to apply complex biological fluids to an immobilized dry strip gel, cast home-made gradient acrylamide gels, run the gels, and perform standard staining methods. A troubleshooting guide is also included. PMID- 17406313 TI - Preparation and use of Coppersensor-1, a synthetic fluorophore for live-cell copper imaging. AB - Coppersensor-1 (CS1) is a small-molecule, membrane-permeable fluorescent dye for imaging labile copper pools in biological samples, including live cells. This probe, comprising a boron dipyrromethene (BODIPY) chromophore coupled to a thioether-rich receptor, has a picomolar affinity for Cu+ with high selectivity over competing cellular metal ions. CS1 fluorescence increases up to 10-fold on binding to Cu+. In this protocol we describe the synthesis of CS1 and how to use this chemical tool to investigate intracellular levels of labile copper in cultured cells. The preparation of CS1 is anticipated to take 4-5 d, and imaging assays can be performed in 1-2 d with cultured cells. PMID- 17406314 TI - Alignment and calibration of a focal neurotransmitter uncaging system. AB - Photolysis of caged compounds is a powerful tool for studying subcellular physiological functions. Here we describe protocols for the alignment and calibration of a focal uncaging system. We also report procedures for convenient quantitative calibration of uncaging. Using these methods, we can achieve submicron lateral resolution of photolysis and probe biological function in spines, the smallest signaling compartments of neurons. Initially, the entire alignment procedure takes 4-6 h to perform; periodic fine-tuning of the system takes 1-2 h. PMID- 17406315 TI - A rat model for neural circuitry abnormalities in schizophrenia. AB - Animal models for complex brain disorders, such as schizophrenia, are essential for the interpretation of postmortem findings. These models allow empirical testing of hypotheses regarding the role of genetic and environmental factors, the pathophysiological mechanisms and brain circuits that are responsible for specific neural abnormalities and their associated behavioral impairment, and the effectiveness of therapeutic treatments relative to these diseases. Recently, we developed a rodent model for neural circuitry abnormalities in discrete corticolimbic subregions of subjects with major psychoses. According to our protocol, the GABA-A receptor antagonist picrotoxin is stereotaxically infused in the basolateral amygdala to mimic a GABA defect in this region that is postulated to occur in these disorders. This protocol has been tested with a number of acute and chronic time schedules. Following picrotoxin administration in the basolateral amygdala, changes in GABAergic neurons and/or terminals in hippocampal regions CA2/3 are observed, similar to those seen in major psychoses, as well as a marked reduction in GABA-receptor-mediated currents in pyramidal neurons of this region. This has established the construct and predictive validity of this model for studying limbic-lobe circuitry abnormalities. We propose that this modeling strategy may provide a valid alternative to isomorphic models of these diseases. PMID- 17406316 TI - A guinea pig model of acute and chronic asthma using permanently instrumented and unrestrained animals. AB - To investigate mechanisms underlying allergen-induced asthmatic reactions, airway hyperresponsiveness and remodeling, we have developed a guinea pig model of acute and chronic asthma using unanesthetized, unrestrained animals. To measure airway function, ovalbumin (IgE)-sensitized animals are permanently instrumented with a balloon-catheter, which is implanted inside the pleural cavity and exposed at the neck of the animal. Via an external cannula, the balloon-catheter is connected to a pressure transducer, an amplifier, an A/D converter and a computer system, enabling on-line measurement of pleural pressure (P(pl))-closely correlating with airway resistance-for prolonged periods of time. Using aerosol inhalations, the method has been successfully applied to measure ovalbumin-induced early and late asthmatic reactions and airway hyperresponsiveness. Because airway function can be monitored repeatedly, intra-individual comparisons of airway responses (e.g., to study drug effects) are feasible. Moreover, this model is suitable to investigate chronic asthma and airway remodeling, which occurs after repeated allergen challenges. The protocol for establishing this model takes about 4 weeks. PMID- 17406317 TI - Morris water maze: procedures for assessing spatial and related forms of learning and memory. AB - The Morris water maze (MWM) is a test of spatial learning for rodents that relies on distal cues to navigate from start locations around the perimeter of an open swimming arena to locate a submerged escape platform. Spatial learning is assessed across repeated trials and reference memory is determined by preference for the platform area when the platform is absent. Reversal and shift trials enhance the detection of spatial impairments. Trial-dependent, latent and discrimination learning can be assessed using modifications of the basic protocol. Search-to-platform area determines the degree of reliance on spatial versus non-spatial strategies. Cued trials determine whether performance factors that are unrelated to place learning are present. Escape from water is relatively immune from activity or body mass differences, making it ideal for many experimental models. The MWM has proven to be a robust and reliable test that is strongly correlated with hippocampal synaptic plasticity and NMDA receptor function. We present protocols for performing variants of the MWM test, from which results can be obtained from individual animals in as few as 6 days. PMID- 17406318 TI - Flow cytometry after bromodeoxyuridine labeling to measure S and G2+M phase durations plus doubling times in vitro and in vivo. AB - This protocol describes methods for calculating the proliferative parameters of cell populations. The basis of the technique is to label cells, either in vitro or in vivo, with halogenated thymidine analogs, such as bromodeoxyuridine (BrdU). Bivariate DNA-BrdU flow cytometry is used to analyze the BrdU-labeled and unlabeled cells. The enumeration of specific cohorts of cells that either have or have not divided in the interval between labeling and cell/tissue sampling permits the calculation of the potential doubling time (T(pot)) of the population, plus the durations of DNA synthesis (T(S)) and the G2+M phase (T(G2+M)) of the cell cycle. The method provides information that is not otherwise available, namely inhibition of DNA synthesis and the separate evaluation of cell-cycle effects in BrdU-labeled and unlabeled subpopulations. Ethanol-fixed samples take 1 d to prepare and stain, and reliable parameter estimates might be obtained from measurements made at a single time point after labeling. PMID- 17406319 TI - Isolation of DNA from bacterial samples of the human gastrointestinal tract. AB - The human gastrointestinal (GI) tract contains a complex microbial community that develops in time and space. The most widely used approaches to study microbial diversity and activity are all based on the analysis of nucleic acids, DNA, rRNA and mRNA. Here, we present a DNA isolation protocol that is suitable for a wide variety of GI tract samples, including biopsies with minute amounts of material. The protocol is set up in such a way that sampling can be performed outside the laboratory, which offers possibilities for implementation in large intervention studies. The DNA isolation is based on mechanical disruption, followed by isolation of nucleic acids using phenol:chloroform:isoamylalcohol extraction. In addition, it includes an alternative DNA isolation protocol that is based on a commercial kit. These protocols have all been successfully used in our laboratory, resulting in isolation of DNA of sufficient quality for microbial diversity studies. Depending on the number of samples and sample type, the whole procedure will take approximately 2.5-4 hours. PMID- 17406320 TI - Cervical duct cannulation in sheep for collection of afferent lymph dendritic cells from head tissues. AB - Pseudo-afferent cervical lymph-duct cannulation in a sheep model allows large amounts of lymph cells to be collected under physiological conditions, carrying immune signaling information from the head tissues, including oro-nasal mucosae. Importantly, large quantities of dendritic cells (DCs) of several subtypes are obtained (up to 8 million per overnight collection), as well as many other trafficking leukocytes. The technique includes three steps: removal of all head lymph nodes on one side (2 h), catheterization of cervical lymph ducts after 2 months (2-3 h) and collection/purification of lymph-cell subsets (4 h). The approach is challenging (1 in 3 success rate) but fruitful, and can be used to study DC subsets under immunomodulation, in order to assess lymph-cell subset dynamic changes and antigen transportation from oro-nasal tissues. This protocol is directed to experienced postdoctoral researchers. PMID- 17406321 TI - Automated serum peptide profiling. AB - Blood is a convenient source of biomarkers. Readily obtainable, it immerses most tissues in the body and is therefore likely to contain cell-derived proteins and peptides that may provide information about various biological processes. Serum proteome and peptidome profiling--using mass spectrometry (MS), for example--may thus show a functional correlate of biological events and disorders. To this end, serum peptides must be enriched and interfering substances removed: a step that should be automated to a degree, reproducible and free of bias if it is to generate a test with any future diagnostic potential. The current protocol allows simultaneous analysis of large numbers of peptides using reversed-phase, magnetic particle-assisted sample processing with a matrix-assisted laser desorption/ionization-time of flight MS readout. It may be used for diagnostic or predictive purposes, specifically as an in vitro readout of proteolytic activities with qualitative and quantitative product analysis, and enables profiling of 96 samples in less than 27 h. PMID- 17406322 TI - Clinical and empirical applications of the Rey-Osterrieth Complex Figure Test. AB - The Rey-Osterrieth Complex Figure Test (ROCF), which was developed by Rey in 1941 and standardized by Osterrieth in 1944, is a widely used neuropsychological test for the evaluation of visuospatial constructional ability and visual memory. Recently, the ROCF has been a useful tool for measuring executive function that is mediated by the prefrontal lobe. The ROCF consists of three test conditions: Copy, Immediate Recall and Delayed Recall. At the first step, subjects are given the ROCF stimulus card, and then asked to draw the same figure. Subsequently, they are instructed to draw what they remembered. Then, after a delay of 30 min, they are required to draw the same figure once again. The anticipated results vary according to the scoring system used, but commonly include scores related to location, accuracy and organization. Each condition of the ROCF takes 10 min to complete and the overall time of completion is about 30 min. PMID- 17406323 TI - Construction of marker-free transplastomic tobacco using the Cre-loxP site specific recombination system. AB - Incorporation of a selectable marker gene in the plastid genome is essential to uniformly alter the thousands of genome copies in a tobacco cell. When transformation is accomplished, however, the marker gene becomes undesirable. Here we describe plastid transformation vectors, the method of plastid transformation using tobacco leaves and alternative protocols for marker gene excision with the P1 bacteriophage Cre-loxP site-specific recombination system. Plastid vectors carry a marker gene flanked with directly oriented loxP sites and a gene of interest, which are introduced into plastids by the biolistic process. The transforming DNA integrates into the plastid genome by homologous recombination via plastid targeting sequences. Marker gene excision is accomplished by a plastid-targeted Cre protein expressed from a nuclear gene. Expression may be from an integrated gene introduced by Agrobacterium transformation (Transformation Protocol), by pollination (Pollination Protocol) or from a transient, non-integrated T-DNA (Transient Protocol). Transplastomic plants are obtained in about 3 months, yielding seed after 2 months. The time required to remove the plastid marker and nuclear genes and to obtain seed takes 10-16 months, depending on which protocol is used. PMID- 17406324 TI - Visualization of F-actin and G-actin equilibrium using fluorescence resonance energy transfer (FRET) in cultured cells and neurons in slices. AB - The plasticity of excitatory synapses has conventionally been studied from a functional perspective. Recent advances in neuronal imaging techniques have made it possible to study another aspect, the plasticity of the synaptic structure. This takes place at the dendritic spines, where most excitatory synapses are located. Actin is the most abundant cytoskeletal component in dendritic spines, and thus the most plausible site of regulation. The mechanism by which actin is regulated has not been characterized because of the lack of a specific method for detection of the polymerization status of actin in such a small subcellular structure. Here we describe an optical approach that allows us to monitor F-actin and G-actin equilibrium in living cells through the use of two-photon microscopy to observe fluorescence resonance energy transfer (FRET) between actin monomers. Our protocol provides the first direct method for looking at the dynamic equilibrium between F-actin and G-actin in intact cells. PMID- 17406325 TI - A protocol for PAIR: PNA-assisted identification of RNA binding proteins in living cells. AB - All aspects of RNA metabolism are regulated by RNA-binding proteins (RBPs) that directly associate with the RNA. Some aspects of RNA biology such as RNA abundance can be readily assessed using standard hybridization technologies. However, identification of RBPs that specifically associate with selected RNAs has been more difficult, particularly when attempting to assess this in live cells. The peptide nucleic acid (PNA)-assisted identification of RBPs (PAIR) technology has recently been developed to overcome this issue. The PAIR technology uses a cell membrane-penetrating peptide (CPP) to efficiently deliver into the cell its linked PNA oligomer that complements the target mRNA sequence. The PNA will then anneal to its target mRNA in the living cell, and then covalently couple to the mRNA-RBP complexes subsequent to an ultraviolet (UV) cross-linking step. The resulting PNA-RNA-RBP complex can be isolated using sense oligonucleotide magnetic beads, and the RBPs can then be identified by mass spectrometry (MS). This procedure can usually be completed within 3 d. The use of the PAIR procedure promises to provide insight into the dynamics of RNA processing, transport, degradation and translation in live cells. PMID- 17406326 TI - Color-coded fluorescence imaging of tumor-host interactions. AB - Fluorescent proteins have the properties of being very bright with high quantum yield and are available in many colors. Tumor-host models consist of transgenic mice expressing green fluorescent protein (GFP) in essentially all cells and tissues or expressing GFP selectively in specific tissues such as blood vessels. Particularly useful are the corresponding nude mice transgenic for GFP expression, as they can accept human tumors. When tumor cells expressing red fluorescent protein are implanted in mice expressing GFP, various types of tumor host interactions can be observed, including those involving host blood vessels, lymphocytes, tumor-associated fibroblasts, macrophages, dendritic cells and others. The 'color-coded' tumor-host models enable imaging and therefore a deeper understanding of the host cells involved and their function in tumor progression. Approximately 4-8 weeks are needed for these procedures. PMID- 17406327 TI - Housing, husbandry and handling of rodents for behavioral experiments. AB - Most animals used in research are rodents, mainly mice because of their predominance in genetics and molecular biology. This article attempts to provide an introduction to mice and rats: health considerations (of the experimenter); choice of species, age, strain and sex; housing and environmental enrichment; and animal identification, handling and dosing. These considerations apply to animal work in general; the rest of the article focuses on the preliminary aspects of behavioral testing, including a protocol for an open field test. This procedure is traditionally associated with activity measurements, and although automated versions are readily available these days, the latter are expensive and may be unavailable in many non-behavioral departments. Moreover, particularly when testing novel genetically modified animals or pharmacological agents, there is no substitute for direct visual observation to detect abnormal signs in the animals: for example, ptosis, piloerection, tremor, ataxia or exophthalmos. The open field test can be adapted in several ways: to assess general behavior and activity (similar to a primary screen in the pharmaceutical industry) or to measure memory (habituation) or anxiety. PMID- 17406328 TI - Chromophore-assisted light inactivation (CALI) using the phototoxic fluorescent protein KillerRed. AB - The phototoxic red fluorescent GFP-like protein KillerRed has recently been described. The phototoxicity of KillerRed exceeds that of EGFP by at least 1,000 fold, making it the first fully genetically encoded photosensitizer. KillerRed opens up new possibilities for precise light-induced cell killing and target protein inactivation. Because KillerRed is encoded by a gene, it can be expressed in a spatially and temporally regulated manner, under a chosen promoter, and fused with the desired protein of interest or localization signal. Here we provide a protocol for target protein inactivation in cell culture using KillerRed. As KillerRed is a new tool, the protocol focuses on aspects that will allow users to maximize the potential of this protein, guiding the design of chimeric constructs, recommended control experiments and preferred illumination parameters. The protocol, which describes target protein visualization and subsequent inactivation, is a 2- or 3-d procedure. PMID- 17406329 TI - Isolation of RNA from bacterial samples of the human gastrointestinal tract. AB - The human gastrointestinal (GI) tract contains a complex microbial community that consists of numerous uncultured microbes. Therefore, nucleic-acid-based approaches have been introduced to study microbial diversity and activity, and these depend on the proper isolation of DNA, rRNA and mRNA. Here, we present an RNA isolation protocol that is suitable for a wide variety of GI tract samples. The procedure for isolating DNA from GI tract samples is described in another Nature Protocols article. One of the benefits of our RNA isolation protocol is that sampling can be performed outside the laboratory, which offers possibilities for implementation in large intervention studies. The RNA isolation is based on mechanical disruption, followed by isolation of nucleic acids using phenol:chloroform:isoamylalcohol extraction and removal of DNA. In our laboratory, this protocol has resulted in the isolation of rRNA and mRNA of sufficient quality and quantity for microbial diversity and activity studies. Depending on the number of samples, the sample type and the quenching procedure chosen, the whole procedure can be performed within 2.5-4 h. PMID- 17406330 TI - Cell tracking using a photoconvertible fluorescent protein. AB - The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confocal laser scanning microscopy to track labeled cells in a pattern of interest in zebrafish embryos. This technique allows the visualization of cell movements and the tracing of neuronal shapes. We provide illustrative examples of expression by mRNA injection, mosaic expression by DNA injection, and the creation of permanent transgenic fish with the UAS-Gal4 system to visualize morphogenetic processes such as neurulation, placode formation and navigation of early commissural axons in the hindbrain. The procedure can be adapted to other photoconvertible and reversible fluorescent molecules, including KikGR and Dronpa; these molecules can be used in combination with two-photon confocal microscopy to specifically highlight cells buried in tissues. The total time needed to carry out the protocol involving transient expression of Kaede by injection of mRNA or DNA, photoconversion and imaging is 2-8 d. PMID- 17406331 TI - Assay of protein kinases using radiolabeled ATP: a protocol. AB - Protein kinase activity results in the incorporation of radiolabeled phosphate from [gamma-32P]ATP into a peptide or protein substrate. The measurement of the amount of radioactivity incorporated into a substrate as a function of time and enzyme concentration allows enzyme activity to be quantified. The activity is expressed as a 'unit', where 1 unit corresponds to the amount of protein kinase that catalyzes the incorporation of 1 nanomole of phosphate into the standard substrate in 1 minute. Specific activity is defined as units of activity per milligram protein. The assay format described here is quick, simple, inexpensive, sensitive and accurate, provides a direct measurement of activity and remains the 'gold standard' for the quantification of protein kinase activity. Up to 40 samples can be assayed manually at one time, and the assay takes one person less than 1 hour to complete. PMID- 17406332 TI - Conjugation of DOTA-like chelating agents to peptides and radiolabeling with trivalent metallic isotopes. AB - Peptides can be labeled with various trivalent radiometals for imaging or targeted radionuclide-therapy applications. The peptide is first conjugated to a chelating agent that is able to form stable complexes with the radionuclide of interest. This conjugation step can be carried out as part of the solid-phase peptide synthesis, or it can be undertaken in the solution phase after synthesis and purification of the peptide. The latter route, described here, involves reacting a molar excess of the activated tri-tert-butyl ester-derivatized chelator with a designated free amino group of a peptide analog, in which all other reactive amines are protected, in the presence of a coupling agent. The conjugate molecule is then purified prior to deprotection and further purification by HPLC. The product can be radiolabeled by addition of a suitable metal salt, followed, if necessary, by removal of the unchelated metal. The entire process of conjugation, purification and radiolabeling should take approximately 12.5 h. PMID- 17406333 TI - Biolistic transfection of neuronal cultures using a hand-held gene gun. AB - Biolistic transfection is a technique in which subcellular-sized particles coated with DNA are accelerated to high velocity to propel them into cells. This method is applicable to tissues, cells and organelles, and can be used for both in vitro and in vivo transformations; with the right equipment, it is simple, rapid and efficient. Here we provide a detailed protocol for biolistic transfection of plasmids into cultured human embryonic kidney (HEK) 293 cells and organotypic brain slices using a hand-held gene gun. There are three major steps: (i) coating microcarriers with DNA, (ii) transferring the microcarriers into a cartridge to make a 'bullet', and (iii) firing the DNA-coated microcarriers into cells using a pulse of helium gas. The method can be readily adapted to other cell types and tissues. The protocol can be completed in 1-2 h. PMID- 17406334 TI - Isolation and purification of afferent lymph dendritic cells that drain the skin of cattle. AB - Dendritic cells (DCs) are central to the induction of immune responses and are a pivotal control point that determines the outcome of infectious challenge. Cannulation of afferent lymphatic vessels allows the isolation of large numbers of lymph DCs. First, lymph nodes that are draining the skin are surgically removed (takes approximately 1 h). Over a period of 6-8 weeks, afferent lymphatic vessels re-anastomose with the efferent duct, forming larger 'pseudoafferent' lymphatic vessels that can be surgically cannulated. Surgical cannulation takes 2 h to perform; daily maintenance of the catheter requires 30 min. Isolation of lymph cells requires 1 h and an additional 60-180 min to enrich or purify the DCs. The lymph can be harvested for up to 1 month, with relatively constant cell numbers and subset distribution throughout this period. This technique, although technically demanding, facilitates studies of DCs and other cells that traffic in the lymph in both the steady state and following antigenic exposure. PMID- 17406335 TI - An ultrastructural readout of fluorescence recovery after photobleaching using correlative light and electron microscopy. AB - Fluorescence recovery after photobleaching (FRAP) provides an important quantitative readout of the mobility of fluorescently tagged structures in live tissue. Here we present a protocol for visualizing FRAP signal at the ultrastructural level, permitting the nature of recovered fluorescence signal to be studied at greater resolution than afforded by conventional light microscopy. Specifically we use FRAP, fixation, photoconversion and correlative light and electron microscopy (CLEM) to examine the ultrastructural organization of mobile FM1-43-labeled vesicles in synapses of cultured hippocampal neurons. At photobleached synapses, the FRAP signal can be visualized as photoconverted electron-dense vesicles. The combination of FRAP and CLEM provides a powerful tool for examining the specific localization of imported vesicles in relation to synaptic architecture. Moreover, with the increasing availability of photoconvertible fluorophores, this approach should be readily applicable to other systems where an ultrastructural characterization of FRAP signal is desirable. After cultures are prepared and ready to use, this protocol takes 2-3 days. PMID- 17406336 TI - Dendrimer-like DNA-based fluorescence nanobarcodes. AB - A major challenge in clinical diagnostics and environmental analysis is the difficulty in rapid and sensitive detection of multiple target molecules simultaneously (i.e., multiplexed detections). Our group has designed and synthesized a dendrimer-like DNA (DL-DNA) that is multivalent and anisotropic; using this unique DNA structure, we have developed a fluorescence-tagged nanobarcode system for multiplex detection. This nanobarcode system allows the rapid and sensitive detection of multiple pathogens simultaneously using the ratios of two different fluorescent dyes, green and red, with which different DL DNAs are labeled. The key step of our nanobarcode model lies in the monodisperse preparation of DL-DNA. Two methods, solution phase and solid phase, are presented here. With slight modifications, this platform technology can also be extended to the multiplexed detection of RNA and proteins. This protocol can be completed in 2-5 d. PMID- 17406337 TI - Studying the uptake of cell-penetrating peptides. AB - More than a decade ago, it was discovered that cationic peptides could traverse the cellular plasma membrane without specific transporter proteins or membrane damage. Subsequently, it was found that these peptides, known as cell-penetrating peptides (CPPs), were also capable of delivering cargos into cells, hence the great potential of these vectors was acknowledged. Today, many different research groups are working with CPPs, which necessitates efforts to develop unified assays enabling the comparison of data. Here we contribute three protocols for evaluation of CPPs which, if used in conjunction, provide complementary data about the amount and mechanism of uptake (fluorometric analysis and confocal microscopy, respectively), as well as the extent of degradation (HPLC analysis of cell lysates). All three protocols are based on the use of fluorescently labeled peptides and can be performed on the same workday. PMID- 17406338 TI - Production of recombinant protein in Pichia pastoris by fermentation. AB - This protocol is applicable to recombinant protein expression by small-scale fermentation using the Pichia pastoris expression system. P. pastoris has the capacity to produce large quantities of protein with eukaryotic processing. Expression is controlled by a methanol-inducible promoter, which allows a biomass generation phase before protein production is initiated. The target protein is secreted directly into a protein-free mineral salt medium, and is relatively easy to purify. The protocol is readily interfaced with expanded bed adsorption for immediate capture and purification of recombinant protein. The setting up of the bioreactor plus the fermentation itself takes 1 wk. Making the master and user seed lots takes approximately 2 wk for each individual clone. PMID- 17406339 TI - A double-reporter splicing assay for determining splicing efficiency in mammalian cells. AB - Changes in alternative splicing patterns can result from both inherited and acquired defects, and they are increasingly recognized as causes of human diseases. Hence, improvements in the understanding of alternative splicing regulation may provide opportunities for restoring productive patterns of splicing. The identification of factors (such as proteins, nucleic acids or small molecules) that modulate the splicing pattern would be facilitated by systems with which many samples can be screened. The absence of reliable systems prompted us to develop an assay system based on dual enzymatic activities. Two distinct signals derived from spliced and unspliced RNA are measured, providing the basis for a robust, rapid and convenient assay for investigating splicing. This protocol describes how to use this system; the time required for lysing the cells and recording enzymatic activity is about 2 hours. PMID- 17406340 TI - Multiplexed absolute quantification for proteomics using concatenated signature peptides encoded by QconCAT genes. AB - An important area of proteomics involves the need for quantification, whether relative or absolute. Many methods now exist for relative quantification, but to support biomarker proteomics and systems biology, absolute quantification rather than relative quantification is required. Absolute quantification usually involves the concomitant mass spectrometric determination of signature proteotypic peptides and stable isotope-labeled analogs. However, the availability of standard labeled signature peptides in accurately known amounts is a limitation to the widespread adoption of this approach. We describe the design and synthesis of artificial QconCAT proteins that are concatamers of tryptic peptides for several proteins. This protocol details the methods for the design, expression, labeling, purification, characterization and use of the QconCATs in the absolute quantification of complex protein mixtures. The total time required to complete this protocol (from the receipt of the QconCAT expression plasmid to the absolute quantification of the set of proteins encoded by the QconCAT protein in an analyte sample) is approximately 29 d. PMID- 17406341 TI - Single- and dual-parameter FRET kinase probes based on pleckstrin. AB - Here we describe protocols for preparing and using fluorescent probes that respond to conformational changes by altered Foerster resonance energy transfer (FRET) efficiencies upon phosphorylation or, in principle, other posttranslational modifications (PTMs). The sensor protein, a truncated version of pleckstrin, is sandwiched between short-wavelength-excitation green fluorescent protein (GFP2) and yellow fluorescent protein (EYFP). As a result of complex conformational changes of the protein upon phosphorylation, the introduction of a second PTM consensus sequence bestows sensitivity to a second modification and yields a dual-parameter probe. The first phase of the protocol lays out the cloning strategy for single- and dual-parameter FRET sensors, including the construction of a versatile platform into which different consensus sequences may be inserted to create diverse probes. Protocols for fluorescence microscopy of the probes in living cells and image processing are also described. Probe preparation takes 7 d; microscopy and image processing take 2 h. PMID- 17406342 TI - Prostate cancer gene therapy clinical trials. AB - Despite recent advances in early detection and treatment, prostate cancer is still the second leading cause of cancer death in men in the United States, and approximately 27,000 men will die from it this year. Better treatments are needed for aggressive forms of localized disease and hormone-refractory metastatic disease. Recently, several gene therapy strategies have generated provocative results in early-stage clinical trials, raising the possibility that gene therapy may have the potential to affect both localized and metastatic disease. Much work lies ahead. Nevertheless, for the time being, these studies provide hope that gene therapy may someday earn a place in the management of prostate cancer. PMID- 17406343 TI - Safety and efficacy of a lentiviral vector containing three anti-HIV genes--CCR5 ribozyme, tat-rev siRNA, and TAR decoy--in SCID-hu mouse-derived T cells. AB - Gene therapeutic strategies show promise in controlling human immunodeficiency virus (HIV) infection and in restoring immunological function. A number of efficacious anti-HIV gene constructs have been described so far, including small interfering RNAs (siRNAs), RNA decoys, transdominant proteins, and ribozymes, each with a different mode of action. However, as HIV is prone to generating escape mutants, the use of a single anti-HIV construct would not be adequate to afford long range-viral protection. On this basis, a combination of highly potent anti-HIV genes--namely, a short hairpin siRNA (shRNA) targeting rev and tat, a transactivation response (TAR) decoy, and a CCR5 ribozyme--have been inserted into a third-generation lentiviral vector. Our recent in vitro studies with this construct, Triple-R, established its efficacy in both T-cell lines and CD34 cell derived macrophages. In this study, we have evaluated this combinatorial vector in vivo. Vector-transduced CD34 cells were injected into severe combined immunodeficiency (SCID)-hu mouse thy/liv grafts to determine their capacity to give rise to T cells. Our results show that phenotypically normal transgenic T cells are generated that are able to resist HIV-1 infection when challenged in vitro. These important attributes of this combinatorial vector show its promise as an excellent candidate for use in human clinical trials. PMID- 17406344 TI - Thermosensitive hydrogel PEG-PLGA-PEG enhances engraftment of muscle-derived stem cells and promotes healing in diabetic wound. AB - Regenerating new tissue using cell transplantation has relied on successful cell engraftment in the host; however, cell engraftment into the diabetic skin wound is not as successful as in many other tissues. We used a biodegradable and biocompatible triblock co-polymer poly(ethylene glycol-b-[DL-lactic acid-co glycolic acid]-b-ethylene glycol) (PEG-PLGA-PEG), which forms a thermosensitive hydrogel, as a wound dressing and scaffold. We found that the thermosensitive hydrogel increased the engraftment of muscle-derived stem cells (MDSCs) by 20- to 30-fold until day 20, when the wound was completely closed in a db/db genetically diabetic mouse model. At day 9, 30% of the transplanted MDSCs were found to remain, and 15% remained at day 20 after transplantation. The increased engraftment resulted in enhanced wound healing, as indicated by the wound closure rate, epithelium migration, and collagen deposition. Using MDSCs stably expressing beta-gal and immunofluorescence, we found that 25% of MDSCs differentiated into fibroblasts, 10% into myofibroblasts, and 10% into endothelial cells. We conclude that using the thermosensitive hydrogel as a scaffold increased the engraftment of MDSCs, which leads to improved diabetic wound healing, possibly by retaining the cells at the wound site for longer. PMID- 17406345 TI - Improving transcriptional termination of self-inactivating gamma-retroviral and lentiviral vectors. AB - Adverse events relating to insertional mutagenesis have reinforced the interest in self-inactivating (SIN) gamma-retroviral and lentiviral vectors without enhancer-promoter sequences in the U3 region of the long terminal repeats. However, SIN vectors suffer from leaky transcriptional termination, increasing the probability of read-through into cellular genes. To improve 3' end processing, we incorporated seven upstream polyadenylation enhancer elements (or upstream sequence elements, USEs) derived from viral or cellular genes into the 3' U3 region of gamma-retroviral and lentiviral SIN vectors. A 100-base-pair sequence representing a recombinant direct repeat of the USE derived from simian virus 40 (2xSV USE) gave the best results, improving both titer and gene expression. In both gamma-retroviral and lentiviral SIN vectors, the 2xSV USE partially substituted for effects provided by the much larger post transcriptional regulatory element derived from woodchuck hepatitis virus (wPRE). By northern blot and reporter assays, we found that the 2xSV USE greatly improved proper messenger RNA (mRNA) processing at the retroviral termination signal. Importantly, the 2xSV USE was superior to the wPRE in suppressing transcriptional read-through, improving not only vector efficiency but potentially also biosafety. PMID- 17406346 TI - Metabolic basis of sexual dimorphism in PKU mice after genome-targeted PAH gene therapy. AB - We have previously reported a transgene delivery system based on phiBT1 bacteriophage integrase that results in targeted insertion of transgenes into mammalian genomes, and its use in the delivery of murine phenylalanine hydroxylase (PAH) complementary DNA (cDNA) into the hepatocytes of male phenylketonuria (PKU) mice, leading to a complete and permanent correction of their hyperphenylalaninemic phenotype. In this study, we report only partial phenotypic correction in female PKU mice, even though hepatic PAH activities in both sexes after gene treatment were similar. Daily injections of tetrahydrobiopterin (BH4), an essential co-factor for phenylalanine hydroxylation, in the gene-treated females led to complete correction of their PKU phenotype. After gonadectomy, serum phenylalanine levels in the gene-treated females were reduced to normal, whereas those in the gene-treated males remained unchanged. The sterile gene-treated PKU mice were subjected to daily sex hormone injections. Whereas the estradiol-treated sterile males developed hyperphenylalaninemia, the dihydrotestosterone-treated sterile females remained normal phenylalaninemic. The results indicate that it is estrogen that suppresses the steady-state levels of BH4 in mouse hepatocytes that became limiting, which is the underlying mechanism for the observed sexual dimorphism in PKU mice after PAH gene treatment. Livers of the PAH gene-corrected PKU mice also appeared normal and without apparent pathologies. PMID- 17406347 TI - Have we been guilty of ageism in the primary treatment of breast cancer? PMID- 17406348 TI - Brief smoking cessation advice from practice nurses during routine cervical smear tests appointments: a cluster randomised controlled trial assessing feasibility, acceptability and potential effectiveness. AB - The aim of this study is to assess the potential effectiveness, acceptability and feasibility of a brief smoking cessation intervention delivered as part of cervical screening. A cluster randomised controlled trial was conducted with clinic week as the unit of randomisation, comparing a group (n=121) receiving brief smoking cessation advice supplemented with written information given by practice nurses during cervical smear test appointments, with a group (n=121) not receiving this advice. Outcomes were intention to stop smoking (potential effectiveness); intention to attend for future cervical screening (acceptability); duration of intervention (feasibility). 172/242 (71%) and 153/242 (63%) participants completed 2-week and 10-week follow-ups, respectively. Compared to women in the control group, those in the intervention group had higher intentions to stop smoking at 2-weeks (adjusted mean difference 0.51, 95% CI: -0.02 to 1.03, P=0.06) and 10-weeks (adjusted mean difference 0.80, 95% CI 0.10 to 1.50, P=0.03). The two groups had similarly high intentions to attend for future screening. Consultations in the intervention arm took a mean of 4.98 min (95% CI: 3.69 to 6.27; P<0.001) longer than the control arm. In conclusion, brief smoking cessation advice given by practice nurses as part of cervical screening seems acceptable, feasible and potentially effective. Evidence is lacking on the effectiveness and cost effectiveness of this intervention in achieving biochemically validated smoking cessation. PMID- 17406349 TI - Worldwide variation in the relative importance of hepatitis B and hepatitis C viruses in hepatocellular carcinoma: a systematic review. AB - We combined information published worldwide on the seroprevalence of hepatitis B surface antigen (HbsAg) and antibodies against hepatitis C virus (anti-HCV) in 27 881 hepatocellular carcinomas (HCCs) from 90 studies. A predominance of HBsAg was found in HCCs from most Asian, African and Latin American countries, but anti-HCV predominated in Japan, Pakistan, Mongolia and Egypt. Anti-HCV was found more often than HBsAg in Europe and the United States. PMID- 17406350 TI - Predicted trends in long-term breast cancer survival in England and Wales. AB - Trends in long-term relative survival from breast cancer are examined for women diagnosed in England and Wales up to 2001, using both period and hybrid approaches. Large improvements in long-term survival are predicted. Women with breast cancer still experience persistent excess mortality up to at least 20 years after diagnosis. PMID- 17406351 TI - Meat consumption and risk of breast cancer in the UK Women's Cohort Study. AB - We performed a survival analysis to assess the effect of meat consumption and meat type on the risk of breast cancer in the UK Women's Cohort Study. Between 1995 and 1998 a cohort of 35 372 women was recruited, aged between 35 and 69 years with a wide range of dietary intakes, assessed by a 217-item food frequency questionnaire. Hazard ratios (HRs) were estimated using Cox regression adjusted for known confounders. High consumption of total meat compared with none was associated with premenopausal breast cancer, HR=1.20 (95% CI: 0.86-1.68), and high non-processed meat intake compared with none, HR=1.20 (95% CI: 0.86-1.68). Larger effect sizes were found in postmenopausal women for all meat types, with significant associations with total, processed and red meat consumption. Processed meat showed the strongest HR=1.64 (95% CI: 1.14-2.37) for high consumption compared with none. Women, both pre- and postmenopausal, who consumed the most meat had the highest risk of breast cancer. PMID- 17406354 TI - The p53 codon 72 proline allele is endowed with enhanced cell-death inducing potential in cancer cells exposed to hypoxia. AB - The preferential retention of the arginine allele at the p53 codon 72 locus is commonly observed in tumours from arginine/proline heterozygotes. Considering that cancer cells are harboured in a hypoxic environment in vivo, we here tested the hypothesis that the p53 codon 72 proline allele confers a survival disadvantage in presence of hypoxia. Here, we show that the transient transfection of the proline allele in p53 null cancer cells exposed to low oxygen tension or to the hypoxia-mimetic drug Desferoxamine induces a higher amount of cell death than the arginine allele. Accordingly, proline allele transiently transfected cell lines express lower levels of hypoxia pro-survival genes (HIF 1alpha, carbonic anhydrase IX, vascular endothelial growth factor, heme oxygenase I, hepatocyte growth factor receptor, vascular endothelial growth factor receptor 2), compared to those transiently transfected with the arginine allele. Further, we report that the exposure of the arginine/proline heterozygote MCF-7 breast cancer cell line to cytotoxic concentration of Desferoxamine for several weeks, gives raise to hypoxia-resistant clones, carrying the arginine, but not the proline allele. These data indicate that the p53 codon 72 proline allele is less permissive for the growth of cancer cells in a hypoxic environment, and suggest that the preferential retention of the arginine allele in the tumour tissues of arginine/proline heterozygous patients may depend upon its lowered capacity to induce cell death in a hypoxic tumour environment. PMID- 17406355 TI - Familial risk for gastric carcinoma: an updated study from Sweden. AB - Reliable data on familial risks are important for clinical counselling and cancer genetics. However, the estimates of familial risk of gastric cancer vary widely. We examined the risk of familial gastric cancer using the updated Swedish Family Cancer Database with 5358 patients among offspring and 36,486 patients among parents. There were 133 families with one parent and one offspring diagnosed with gastric cancer, and 20 families with two affected offspring. Familial standardised incidence ratios (SIRs) were 1.63 and 2.93 when parents and siblings presented with gastric cancer, respectively. The high sibling risk was owing to cancer in the corpus (SIR 7.28). The SIR for cardia cancer was 1.54 when parents were diagnosed with any gastric cancer. Cardia cancer associated with oesophageal cancer, particularly with oesophageal adenocarcinoma. Among specific histologies, signet ring cancer showed an increase. A few associations were noted for discordant sites, including the urinary bladder and the endometrium. H. pylori infection, although not measured in the present study, is probably an important risk factor for the high sibling risk of corpus cancer. Familial clustering of cardia cancer is independent of H. pylori infection, and may have a genetic basis. The familial association of cardia cancer with oesophageal adenocarcinoma may provide aetiological clues. PMID- 17406356 TI - Predicting gene promoter methylation in non-small-cell lung cancer by evaluating sputum and serum. AB - The use of 5-methylcytosine demethylating agents in conjunction with inhibitors of histone deacetylation may offer a new therapeutic strategy for lung cancer. Monitoring the efficacy of gene demethylating treatment directly within the tumour may be difficult due to tumour location. This study determined the positive and negative predictive values of sputum and serum for detecting gene methylation in primary lung cancer. A panel of eight genes was evaluated by comparing methylation detected in the primary tumour biopsy to serum and sputum obtained from 72 patients with Stage III lung cancer. The prevalence for methylation of the eight genes in sputum (21-43%) approximated to that seen in tumours, but was 0.7-4.3-fold greater than detected in serum. Sputum was superior to serum in classifying the methylation status of genes in the tumour biopsy. The positive predictive value of the top four genes (p16, DAPK, PAX5 beta, and GATA5) was 44-72% with a negative predictive value for these genes > or =70%. The highest specificity was seen for the p16 gene, and this was associated with a odds ratio of six for methylation in the tumour when this gene was methylated in sputum. In contrast, for serum, the individual sensitivity for all genes was 6 27%. Evaluating the combined effect of methylation of at least one of the four most significant genes in sputum increased the positive predictive value to 86%. These studies demonstrate that sputum can be used effectively as a surrogate for tumour tissue to predict the methylation status of advanced lung cancer where biopsy is not feasible. PMID- 17406357 TI - PTHrP increases transcriptional activity of the integrin subunit alpha5. AB - Increasing evidence is emerging highlighting the role of parathyroid hormone related protein (PTHrP) during metastasis by regulating cell adhesion. The current study demonstrated that modulation of PTHrP expression by PTHrP overexpression and small interfering RNA-induced silencing resulted in changes in cell adhesion and integrin expression. RNA interference of endogenous PTHrP caused a significant reduction in cell adhesion of a breast cancer cell line to collagen type I, fibronectin and laminin (P<0.05) and of a colon cancer cell to collagen type I and fibronectin (P<0.05). Overexpression of PTHrP induced a significant increase in cell adhesion of colon (P<0.0001) and breast (P<0.05) cancer cells to the same extracellular matrix proteins. These PTHrP-mediated effects were attributed to changes in integrin expression as the differences in adhesion profile correlated with the integrin expression profile. In an attempt to elucidate the mechanism whereby PTHrP regulates integrin expression, promoter activity of the integrin alpha5 subunit was analysed and significant increases in transcriptional activity were observed in PTHrP overexpressing cells (P<0.0001), which was dependent on nuclear localisation. These results indicate that modulation of cell adhesion is a normal physiological action of PTHrP, mediated by increasing integrin gene transcription. PMID- 17406358 TI - Systematic review, including meta-analyses, on the management of locally advanced pancreatic cancer using radiation/combined modality therapy. AB - There is no consensus on the management of locally advanced pancreatic cancer, with either chemotherapy or combined modality approaches being employed (Maheshwari and Moser, 2005). No published meta-analysis (Fung et al, 2003; Banu et al, 2005; Liang, 2005; Bria et al, 2006; Milella et al, 2006) has included randomised controlled trials employing radiation therapy. The aim of this systematic review was to compare the following: (i) chemoradiation followed by chemotherapy (combined modality therapy) vs best supportive care (ii) radiotherapy vs chemoradiation (iii) radiotherapy vs combined modality therapy (iv) chemotherapy vs combined modality therapy (v) 5FU-based combined modality treatment vs another-agent-based combined modality therapy. Relevant randomised controlled trials were identified by searching databases, trial registers and conference proceedings. The primary end point was overall survival and secondary end points were progression-free survival/time-to-progression, response rate and adverse events. Survival data were summarised using hazard ratio (HR) and response-rate/adverse-event data with relative risk. Eleven trials involving 794 patients met the inclusion criteria. Length of survival with chemoradiation was increased compared with radiotherapy alone (two trials, 168 patients, HR 0.69; 95% confidence interval (CI) 0.51-0.94), but chemoradiation followed by chemotherapy did not lead to a survival advantage over chemotherapy alone (two trials, 134 patients, HR 0.79; CI 0.32-1.95). Meta-analyses could not be performed for the other comparisons. A survival benefit was demonstrated for chemoradiation over radiotherapy alone. Chemoradiation followed by chemotherapy did not demonstrate any survival advantage over chemotherapy alone, but important clinical differences cannot be ruled out due to the wide CI. PMID- 17406359 TI - Mitogenic growth signalling, DNA replication licensing, and survival are linked in prostate cancer. AB - Activation of mitogen/extracellular-signal-regulated kinase kinase 5/extracellular signal-regulated kinase-5 (MEK5/ERK5) growth signalling is coupled to increased cell proliferation in prostate cancer (PCa). Dysregulation of the DNA replication licensing pathway, a critical step in growth control downstream of transduction signalling pathways, is associated with development of PCa. In this study we have investigated linkages between the MEK5/ERK5 pathway and DNA replication licensing during prostate carcinogenesis. The effects of increased MEK5/ERK5 signalling on the expression of replication licensing factors Mcm2 and geminin and the proliferation marker Ki67 were studied in an ecdysone inducible system expressing a constitutively activated mutant of MEK5 in EcR293 cells and in stable ERK5 over-expressing PC3 clones. In parallel, expression of these biomarkers in PCa biopsy specimens (n=58) was studied and compared to clinicopathological parameters. In both in vitro systems induction of MEK5 expression resulted in increased levels of phosphorylated ERK5 and Mcm2, geminin and Ki67 proteins. In PCa specimens average Mcm2 expression was greater than Ki67 and geminin expression (median labelling index (LI) 36.7, 18.1, and 3.4% respectively), consistent with their differential expression according to growth status (P<0.0001). Mcm2, geminin and Ki67 expression were significantly associated with Gleason grade (P=0.0002, P=0.0003, P=0.004); however there was no link with T or M stage. There was a significant relationship between increasing ERK5 expression and increasing Mcm2 (P=0.003) and Ki67 (P=0.009) expression, with non-significant trends seen with increasing MEK5 expression. There were significant associations between Gleason grade and the number of cells traversing G1 phase (Ki67(LI)-geminin(LI); (P=0.001)), with high ERK5 levels associated with both an increase in replication licensed but non-cycling cells (Mcm2(LI) Ki67(LI); (P=0.01)) and accelerated cell cycle progression (geminin(LI)/Ki67(LI); (P= 0.005)), all indicative of a shift towards increasing proliferative potential. While Mcm2 and Ki67 were both prognostic factors on univariate analysis, only Mcm2 remained an independent prognostic marker on multivariate analysis. Taken together, our data show that induction of MEK5/ERK5 signalling is linked to activation of the DNA replication licensing pathway in PCa, and that the strong prognostic value of MCM proteins may result from their function as relay stations coupling growth regulatory pathways to genome duplication. PMID- 17406360 TI - Comorbidity and survival of Danish breast cancer patients from 1995 to 2005. AB - Comorbid diseases can affect breast cancer prognosis. We conducted a population based study of Danish women diagnosed with a first primary breast cancer from 1995 to 2005 (n=9300), using hospital discharge registry data to quantify comorbidities by Charlson score. We examined the influence of comorbidities on survival, and quantified their impact on relative mortality rates. The prevalence of patients with a Charlson score='0' fell from 86 to 81%, with an increase in those with Charlson score='1-2' from 13 to 16%, and score='3+' from 1 to 2%. One- and five-year survival for patients with Charlson score='0' and '1-2' was better for those diagnosed in 1998-2000 than in 1995-1997. Overall, patients diagnosed in 2001-2004 (mortality ratio (MR)=0.80, 95% CI=0.68-0.95) and 1998-2000 (MR=0.92, 95% CI=0.78-1.09) had lower 1-year age-adjusted mortality compared to those diagnosed in 1995-1997 (reference period). Patients with Charlson scores '1 2' and '3+' had higher age-adjusted 1-year mortality than those with a Charlson score='0' in each time period (2001-2004: MR('1-2')=1.76, 95% CI=1.35-2.30, and MR('3+')=3.78, 95% CI=2.51-5.68; and 1998-2000: MR('1-2')=1.60, 95% CI=1.36-1.88 and MR('3+')=2.34, 95% CI=1.65-3.33). Similar findings were observed for 5-year age-adjusted mortality. Additional analyses, adjusted for stage, indicated that confounding by stage could not explain these findings. Despite continued improvements in breast cancer survival, we found a trend of poorer survival among breast cancer patients with severe comorbidities even after adjusting for age and stage. Such poorer survival is an important public health concern and can be expected to worsen as the population ages. PMID- 17406361 TI - Tumour M2-PK as a stool marker for colorectal cancer: comparative analysis in a large sample of unselected older adults vs colorectal cancer patients. AB - Stool testing based on tumour-derived markers might offer a promising approach for non-invasive colorectal cancer (CRC) screening. The aim of this study was to estimate the potential of a new test for faecal tumour M2-PK to discriminate patients with CRC from a large sample of unselected older adults. Faecal tumour M2-PK concentrations were determined in 65 CRC patients and in a population-based sample of 917 older adults (median age: 65 and 62 years, respectively). Sensitivity and specificity of the test were calculated at different cutoff values, and receiver-operating characteristic curves (ROC) were constructed to visualise the discriminatory power of the test. The median (interquartile range) faecal tumour M2-PK concentration was 8.6 U ml(-1) (2.8-18.0) among CRC patients and <2 U ml(-1) (<2-3.2; P<0.0001) in the population sample. At a cutoff value of 4 U ml(-1), sensitivity (95% confidence interval) was 85% (65-96%) for colon cancer and 56% (41-74%) for rectum cancer. Specificity (95% confidence interval) was estimated to be 79% (76-81%). Given the comparatively high sensitivity of the tumour M2-PK stool test (especially for colon cancer) and its simple analysis, the potential use of the test for early detection of CRC merits further investigation. Possibilities to enhance specificity of the test should be explored. PMID- 17406362 TI - Post-treatment parenthood in Hodgkin's lymphoma survivors. AB - Attempted and achieved post-treatment parenthood, with or without use of assisted reproduction techniques (ARTs), was assessed in Hodgkin's lymphoma survivors treated from 1971-1998, aged below 50 (females) or 65 (males) at diagnosis, aged 18 to 75 at survey. Four treatment groups were constructed: radiotherapy only, low -, medium - and high gonadotoxic chemotherapy (with or without radiotherapy in the three chemotherapy groups). Using Kaplan-Meier estimates, log-rank tests and Cox regression analyses, factors influencing post-treatment parenthood were investigated, with birth of the first child after treatment as the end point. Forty-five per cent (120/269) of males and 50% (91/184) of females reported attempted post-treatment parenthood. Of these, 76 (63%) males and 68 (75%) females had a child without use of ARTs. In addition 10 males and one female achieved post-treatment parenthood with use of ARTs. Treatment group was significantly associated with post-treatment parenthood, with highest probabilities after radiotherapy only and low gonadotoxic chemotherapy. In univariate analyses, age at diagnosis was a significant factor related to post treatment parenthood in females. PMID- 17406363 TI - Helicobacter pylori eradication for low-grade gastric mucosa-associated lymphoid tissue lymphoma is more successful in inducing remission in distal compared to proximal disease. AB - A series of studies has shown that Helicobacter pylori eradication induces remission in most patients with low-grade gastric mucosa-associated lymphoid tissue (MALT) lymphoma. However, there have been few reports about the effect of bacterial treatment on the gastric MALT lymphoma in Korea, a well-known H. pylori endemic area. A total of 111 H. pylori-infected patients were prospectively enrolled in Seoul National University Hospital and 99 among them were completely followed up according to our protocol. After H. pylori eradication, tumoural response was evaluated by endoscopy and histopathology every 2-3 months till complete remission (CR) and every 6 months after achieving CR. Median follow-up period was 41 months (range, 11-125 months). Helicobacter pylori was successfully eradicated in all 99 patients and CR was obtained in 84 (84.8%) of 99 patients. The median time to reach CR was 3 months and 94% of CR is in continuous complete remission. Five patients with CR relapsed after 10-22 months without the evidence of H. pylori reinfection. Cumulative recurrence rate was 2.3, 7.7 and 9.3% at 1, 2 and 3 years, respectively. Tumours were mainly located in distal stomach (67.7%) and tumours in distal stomach were associated with more favourable response than those in proximal stomach (P=0.001). Majority of patients with low grade gastric MALT lymphoma treated by exclusive H. pylori eradication have a favourable long-term outcome, offering a real chance of cure. Tumour location could be a predictive factor for remission following H. pylori eradication. PMID- 17406364 TI - Exploiting nanotechnology to target cancer. AB - Nanotechnology is increasingly finding use in the management of cancer. Nanoscale devices have impacted cancer biology at three levels: early detection using, for example, nanocantilevers or nanoparticles; tumour imaging using radiocontrast nanoparticles or quantum dots; and drug delivery using nanovectors and hybrid nanoparticles. This review addresses some of the major milestones in the integration of nanotechnology and cancer biology, and the future of nanoscale approaches for cancer management. PMID- 17406365 TI - Co-culturing human prostate carcinoma cells with hepatocytes leads to increased expression of E-cadherin. AB - Metastasis is a multi-step process wherein tumour cells detach from the primary mass, migrate through barrier matrices, gain access to conduits to disseminate, and subsequently survive and proliferate in an ectopic site. During the initial invasion stage, prostate carcinoma cells undergo epithelial-mesenchymal-like transition with gain of autocrine signalling and loss of E-cadherin, hallmarks that appear to enable invasion and dissemination. However, some metastases express E-cadherin, and we found close connections between prostate carcinoma cells and hepatocytes in a liver microtissue bioreactor. We hypothesise that phenotypic plasticity occurs late in prostate cancer progression at the site of ectopic seeding. Immunofluorescence staining for E-cadherin in co-cultures of hepatocytes and DU-145 prostate cancer cells revealed E-cadherin upregulation at peripheral sites of contact by day 2 of co-culture; E-cadherin expression also increased in PC-3 cells in co-culture. These carcinoma cells bound to hepatocytes in an E-cadherin-dependent manner. Although the signals by which the hepatocytes elicited E-cadherin expression remain undetermined, it appeared related to downregulation of epidermal growth factor receptor (EGFR) signalling. Inhibition of autocrine EGFR signalling increased E-cadherin expression and cell-cell heterotypic adhesion; further, expression of a downregulation-resistant EGFR variant prevented E-cadherin upregulation. These findings were supported by finding E-cadherin and catenins but not activated EGFR in human prostate metastases to the liver. We conclude that the term epithelial-mesenchymal transition only summarises the transient downregulation of E-cadherin for invasion with re-expression of E-cadherin being a physiological consequence of metastatic seeding. PMID- 17406366 TI - The prognostic role of WHO classification, urinary 5-hydroxyindoleacetic acid and liver function tests in metastatic neuroendocrine carcinomas of the gastroenteropancreatic tract. AB - The World Health Organisation (WHO) classification (2000) is widely used to classify neuroendocrine carcinomas (NECs), yet its prognostic value needs to be confirmed. In this study, patients with metastatic NECs (n=119) were classified according to WHO guidelines into well differentiated and poorly differentiated (WDNECs and PDNECs). Histological differentiation based on WHO criteria had the highest impact on overall survival (OS) (PDNECs : WDNECs hazard ratio (HR)=4.02, P=0.02); however, PDNECs represented only a small percentage of patients (8%). In a WDNEC-restricted analysis, abnormal liver function tests (LFTs) and elevated urinary 5-hydroxyindoleacetic acid (u5HIAA) were independent prognostic factors for survival (HR=2.65, P=0.006 and HR=2.51, P=0.003, respectively) and were used to create a WDNEC-specific prognostic model (low risk=both normal, intermediate risk=one of them abnormal, high risk=both abnormal). Low-risk WDNECs had the most favourable prognosis (median OS, mOS 8.1 years), which was significantly better compared to both intermediate-risk and high-risk WDNECs (mOS 3.2 and 1.4 years, with P=0.01 and P<0.001, respectively). High-risk WDNECs displayed the shortest OS (1.3 years), which was similar to that of PDNECs (P=0.572). This analysis supports the prognostic value of WHO classification for metastatic NECs arising from the gastroenteropancreatic tract; however, risk stratification using readily available u5HIAA and LFTs may be necessary for the heterogeneous group of WDNECs. PMID- 17406367 TI - Telomere uncapping by the G-quadruplex ligand RHPS4 inhibits clonogenic tumour cell growth in vitro and in vivo consistent with a cancer stem cell targeting mechanism. AB - The pentacyclic acridinium methosulfate salt RHPS4 induces the 3'single-stranded guanine-rich telomeric overhang to fold into a G-quadruplex structure. Stabilisation of the latter is incompatible with an attachment of telomerase to the telomere and thus G-quadruplex ligands can effectively inhibit both the catalytic and capping functions of telomerase. In this study, we examined mechanisms underlying telomere uncapping by RHPS4 in uterus carcinoma cells (UXF1138L) with short telomeres and compared the susceptibility of bulk and clonogenic cancer cells to the G-quadruplex ligand. We show that treatment of UXF1138L cells with RHPS4 leads to the displacement of the telomerase catalytic subunit (hTERT) from the nucleus, induction of telomere-initiated DNA-damage signalling and chromosome fusions. We further report that RHPS4 is more potent against cancer cells that grow as colonies in soft agar than cells growing as monolayers. Human cord blood and HEK293T embryonic kidney cell colony forming units, however, were more resistant to RHPS4. RHPS4-treated UXF1138L xenografts had a decreased clonogenicity, showed loss of nuclear hTERT expression and an induction of mitotic abnormalities compared with controls. Although single-agent RHPS4 had limited in vivo efficacy, a combination of RHPS4 with the mitotic spindle poison Taxol caused tumour remissions and further enhancement of telomere dysfunction. PMID- 17406368 TI - Poorly differentiated and anaplastic thyroid carcinomas: chromosomal and oligo array profile of five new cell lines. AB - Information on gene alterations associated to poorly differentiated (PDTC) and anaplastic thyroid carcinomas (ATC) is scarce. Using human cancer cell lines as a tool for gene discovery, we performed a cytogenetic and oligo-array analysis in five new cell lines derived from two PDTC and three ATC. In PDTC we evidenced, as important, the involvement of the MAPK/ERK kinase pathway, and downregulation of a group of suppressor genes that include E-cadherin. In ATC, downregulation of a specific group of oncosuppressor genes was also observed. Our ATC cell lines presented chromosomal markers of gene amplification, and we were able to identify for the first time the nature of the involved amplicon target genes. We found that the main molecular differences between the two cell line types were related to signal transduction pathways, cell adhesion and motility process. TaqMan experiments performed for five amplicon target genes and for two genes, which allowed a clear distinction between ATC and PDTC: CDH13 and PLAU corroborated array results, not only in the cell lines, but also in an additional set of primary 14 PDTC and three ATC. We suggest that our findings may represent new tools for the development of more effective therapies to the hitherto untreatable ATC. PMID- 17406369 TI - Can an evidence-based intervention reduce the rate of bloodstream infections in patients with venous catheters? PMID- 17406370 TI - Can antiheart autoantibodies predict disease risk in asymptomatic relatives of patients with dilated cardiomyopathy? PMID- 17406371 TI - Can dehydroepiandrosterone or testosterone replacement effectively treat the symptoms of aging? PMID- 17406372 TI - Is it worthwhile performing a PPI trial in primary-care patients with GERD symptoms? PMID- 17406373 TI - Is biliopancreatic diversion with duodenal switch the best surgical treatment for super-obese patients? PMID- 17406374 TI - Ultrasonographic findings in shoulders of able-bodied, paraplegic and tetraplegic subjects. AB - STUDY DESIGN: Cross-sectional. OBJECTIVES: To evaluate the association between spinal cord injury (SCI) and ultrasonographic findings in shoulders. SETTING: Randomly selected patients of Kapyla Rehabilitation Centre and able-bodied citizens of Helsinki, Finland. METHODS: Ultrasonography of the shoulder was performed on able-bodied, tetraplegic and paraplegic subjects. Subjects with SCI were selected from patients of Kapyla Rehabilitation Centre, who were injured more than 3 months before the examination. RESULTS: Findings of the glenohumeral joint differed most clearly between the groups. Significant association between SCI level and oedema of the glenohumeral joint was noted in multiple regression analysis, where other background factors were also considered. CONCLUSION: Paraplegic and tetraplegic subjects are prone to glenohumeral changes that can be verified with ultrasonography. PMID- 17406375 TI - The anterior spinal artery origin: a microanatomical study. AB - BACKGROUND: Detailed knowledge of the pattern of origin of the anterior spinal artery is critical as surgical and endovascular procedures involving the area of the ventral medulla and the vertebrobasilar junction are commonplace. We conducted a detailed microanatomical study to elucidate the site and pattern of origin of this critically important artery. METHOD: Nine adult cadaveric heads (18 sides) were examined after injection of colored silicon. In every specimen, the site of origin of the rami of the anterior spinal artery, their course, branching pattern and anastomoses, external diameters, and the distance from neighbor critical vessels were recorded. The dissections were performed with the aid of both the surgical microscope and a 0 degree endoscope. FINDINGS: The pattern and site of origin of the anterior spinal artery show great variability. Also the distance of the origin of the two rami (right and left) forming the anterior spinal artery from the vertex of the vertebrobasilar junction and from the origin of the posterior inferior cerebellar artery is highly variable. CONCLUSIONS: Knowledge of the different pattern of origin and course of the proximal portion of the anterior spinal artery is critically important when planning and executing endovascular and surgical procedures involving the distal vertebral artery, the vertebrobasilar junction and the ventral medulla. On the basis of our and other authors' findings, we propose an overall classification of the pattern of origin and distribution of the proximal anterior spinal artery, which has clinical repercussions. PMID- 17406376 TI - Immunomodulatory effect of the purine nucleoside inosine following spinal cord contusion injury in rat. AB - STUDY DESIGN: In vivo study using a moderate spinal cord contusion injury (SCI) model in adult rat. OBJECTIVE: To assess the immunomodulatory effects of the purine nucleoside inosine on macrophage/microglia activation at and near the lesion site and in white matter areas remote from the injury epicenter. SETTING: Department of Cell and Developmental Biology, SUNY Upstate Medical University, Syracuse, NY, USA. METHODS: Animals (N=56) were injured using a moderate SCI at T9-T10 spinal level and were divided into three groups, depending on treatment paradigm. Rats received either intraperitoneal or subcutaneous injections of inosine (N=28) or vehicle (N=28). Spinal cord tissue was processed for ED-1 immunoreactivity and the volume fraction of ED-1(+) profiles was calculated using the Cavalieri method and unbiased stereology. RESULTS: The volume fraction of ED 1(+) profiles within gray and lateral white matter regions at and around the lesion site was significantly reduced only following a twice daily-6 week treatment course, compared with vehicle controls, and white matter areas remote from the lesion were unaffected by all inosine treatment paradigms. CONCLUSIONS: Continued subcutaneous delivery of inosine, beginning 15-min post-SCI and persisting throughout the survival period of 6 weeks exerted immunomodulatory effects at and around the lesion site. PMID- 17406377 TI - Subacute progressive ascending myelopathy following spinal cord injury: MRI appearances and clinical presentation. AB - STUDY DESIGN: Retrospective Case Review. OBJECTIVES: To describe the clinical presentation and course of patients with magnetic resonance imaging (MRI) features of subacute progressive ascending myelopathy (SPAM). A rare complication of spinal cord injury. SETTING: National Spinal Injuries Centre, Stoke Mandeville Hospital, UK. MATERIALS AND METHODS: A retrospective review of the case notes and MRI studies of 11 cases with typical MRI features of ascending myelopathy presenting to a tertiary Spinal Injuries centre over a 15-year period. RESULTS: Eleven patients were identified with MRI features typical of SPAM, a median of 13 days (mean 24, range 4-86 days) following cord injury. The median number of cord segments involved above the initial insult was 6 (mean 6.2, range 4-11). MRI appearances include extension four or more segments cephalad to the initial cord injured segment, cord expansion and increased intramedullary T2 signal with a rim of cord sparing peripherally. Cord changes and neurological deficits improved over time but did not return to the initial injured level on MRI. One patient died (mortality 9%). CONCLUSION: SPAM is a rare cause of neurological deterioration following spinal cord injury but may remain subclinical. SPAM most commonly presents as neurological deterioration but may present with shoulder pain, respiratory deterioration or remain subclinical. There are characteristic MRI appearances. It can be fatal. PMID- 17406378 TI - Cold pressor test in tetraplegia and paraplegia suggests an independent role of the thoracic spinal cord in the hemodynamic responses to cold. AB - BACKGROUND: Cold application to the hand (CAH) is associated in healthy people with increase in heart rate (HR) and blood pressure (BP). OBJECTIVE: To study hemodynamic responses to CAH in humans following spinal cord injuries of various levels, and examine the effect of spinal cord integrity on the cold pressor response. DESIGN: An experimental controlled study. SETTING: The spinal research laboratory, Loewenstein Hospital, Raanana, Israel. SUBJECTS: Thirteen healthy subjects, 10 patients with traumatic T(4-6) paraplegia and 11 patients with traumatic C(4-7) tetraplegia. MAIN OUTCOME MEASURES: HR, BP, HR and BP spectral components (low frequency, LF; high frequency, HF; LF/HF), cerebral blood flow velocity (CBFV) and cerebrovascular resistance index (CVRi). METHODS: The outcome measures of the three subject groups monitored for HR, BP and CBFV were compared from 5 min before to 5 min after 40-150 s of CAH. The recorded signals were digitized online and analyzed offline in both the time and frequency domains. RESULTS: During CAH, HR and CVRi increased significantly in all subject groups (P<0.001), and BP in control subjects and in the tetraplegia group (P<0.01). BP increase was not statistically significant in paraplegia, and CBFV, HR LF, HR HF and BP LF did not change significantly during CAH in any group. CONCLUSIONS: The CAH effect in tetraplegia and the suppressed BP increase in paraplegia, supported by the other findings, suggest a contribution of an independent thoracic spinal mechanism to the cold pressor response. PMID- 17406379 TI - Clinical correlates of fatigue in spinal cord injury. AB - STUDY DESIGN: Retrospective chart review. OBJECTIVES: To determine the prevalence of fatigue in an outpatient spinal cord injury population and to examine the clinical variables contributing to that fatigue. SETTING: GF Strong Rehabilitation Centre, Vancouver, British Columbia, Canada. METHODS: Medical charts of 76 individuals admitted to the GF Strong Outpatient SCI Program between December 2004 and December 2005 were reviewed. Data collected included information on clinical characteristics, demographics and Fatigue Severity Scale (FSS) scores. Multivariable analysis was completed to determine the independent association between these variables and fatigue severity. RESULTS: A total of 57% (95% confidence interval (CI)=45-67%) of the sample were found to have fatigue severe enough to interfere with function. People that were admitted for medical reasons; had pain, spasticity, incomplete injuries, and/or were on more that one medication with a known side effect of fatigue had significantly higher FSS scores. Multivariable analysis indicated incomplete injury was the only statistically significant predictor of a higher FSS scores; pain approached significance (P=0.07, CI=-0.09, 2.06). Together these variables account for 18% of the variance in FSS scores in this sample. CONCLUSION: Fatigue among individuals with spinal cord injury who are seeking outpatient rehabilitation is very common. The severity of fatigue was greater for individuals with incomplete lesions. Pain was also a potentially important covariate of fatigue. Further research is required to determine what else contributes to fatigue severity beyond these clinical variables as only minimal variance was accounted for in our model. PMID- 17406380 TI - Posterior spinal shortening for paraparesis following vertebral collapse due to osteoporosis. AB - STUDY DESIGN: Retrospective case series. OBJECTIVES: To assess the efficacy of posterior spinal shortening for paraparetic patients following vertebral collapse owing to osteoporosis, especially on instrumentation loosening. SETTING: Department of orthopaedic surgery, Jichi Medical University Hospital and Omiya Medical Center in Japan. METHODS: The clinical records and radiographs of 13 patients with paraparesis following vertebral collapse owing to osteoporosis treated with posterior spinal shortening were retrospectively reviewed to evaluate the usefulness of this method. Assessment of the clinical course was done by direct examination in all cases. Ambulatory ability was divided into four categories. RESULTS: Upon final observation, nine cases were able to walk with a cane or crutch, one case remained in gait training, two cases remained unable to stand and one case with urinary incontinence improved in urinary function. In one case, paralysis deteriorated. Vertebral compression fracture of the end vertebrae that were fixed occurred in three cases complicated with rheumatoid arthritis. CONCLUSION: The posterior spinal shortening can be a choice for treating delayed paraparesis following vertebral collapse owing to osteoporosis. PMID- 17406381 TI - Anemia in patients with chronic kidney disease: defining the optimal hemoglobin target. PMID- 17406382 TI - Does adjunctive lamotrigine provide improved control of primary generalized tonic clonic seizures in children? PMID- 17406383 TI - Which second-line treatment regimen should be used following relapse of platinum sensitive ovarian cancer? PMID- 17406384 TI - Should patients with osteoarthritis be treated with COX2 inhibitors rather than traditional NSAIDs? PMID- 17406385 TI - Does treatment with DMARDs decrease the risk of cardiovascular disease in patients with RA? PMID- 17406386 TI - Is hysterectomy associated with an increase in the symptoms of urinary incontinence? PMID- 17406388 TI - Selection and characterization of large collections of peptide aptamers through optimized yeast two-hybrid procedures. AB - Peptide aptamers are combinatorial proteins that specifically bind intracellular proteins and modulate their function. They are powerful tools to study protein function within complex regulatory networks and to guide small-molecule drug discovery. Here we describe methodological improvements that enhance the yeast two-hybrid selection and characterization of large collections of peptide aptamers. We provide a detailed protocol to perform high-efficiency transformation of peptide aptamer libraries, in-depth validation experiments of the bait proteins, high-efficiency mating to screen large numbers of peptide aptamers and streamlined confirmation of the positive clones. We also describe yeast two-hybrid mating assays, which can be used to determine the specificity of the selected aptamers, map their binding sites on target proteins and provide structural insights on their target-binding surface. Overall, 12 weeks are required to perform the protocols. The improvements on the yeast two-hybrid method can be also usefully applied to the screening of cDNA libraries to identify protein interactions. PMID- 17406387 TI - Measuring calcium signaling using genetically targetable fluorescent indicators. AB - Genetically encoded Ca2+ indicators allow researchers to quantitatively measure Ca2+ dynamics in a variety of experimental systems. This protocol summarizes the indicators that are available, and highlights those that are most appropriate for a number of experimental conditions, such as measuring Ca2+ in specific organelles and localizations in mammalian tissue-culture cells. The protocol itself focuses on the use of a cameleon, which is a fluorescence resonance-energy transfer (FRET)-based indicator comprising two fluorescent proteins and two Ca2+ responsive elements (a variant of calmodulin (CaM) and a CaM-binding peptide). This protocol details how to set up and conduct a Ca2+-imaging experiment, accomplish offline data processing (such as background correction) and convert the observed FRET ratio changes to Ca2+ concentrations. Additionally, we highlight some of the challenges in observing organellar Ca2+ and the alternative strategies researchers can employ for effectively calibrating the genetically encoded Ca2+ indicators in these locations. Setting up and conducting an initial calibration of the microscope system is estimated to take approximately 1 week, assuming that all the component parts are readily available. Cell culture and transfection is estimated to take approximately 3 d (from the time of plating cells on imaging dishes). An experiment and calibration will probably take a few hours. Finally, the offline data workup can take approximately 1 d depending on the extent of analysis. PMID- 17406389 TI - Long-term microfluidic cultures of myotube microarrays for high-throughput focal stimulation. AB - We have developed a microfluidic cell culture method that allows for the formation of linear isolated myotubes organized in a parallel microarray. Attachment and spreading of cells are confined within microtracks of cell adherent proteins separated by a protein-repellent coating. Signaling molecules or other molecules of interest can be focally delivered to the myotubes using heterogeneous microfluidic streams. We have used the method to focally deliver agrin (a molecule implicated as a postsynaptic organizer), which leads to localized acetylcholine receptor clustering. These techniques can be modified to accommodate other cell types and can be adapted to virtually any bioactive molecule such as signaling factors or drugs. This protocol features two major techniques that can be utilized simultaneously or independently to (i) micropattern cells using surface chemical modification and (ii) use a microfluidic platform for culturing and focal stimulation of cells with molecules of interest. Device design, fabrication and assembly can be completed in 3 days. PMID- 17406390 TI - Genetic transformation of tobacco NT1 cells with Agrobacterium tumefaciens. AB - This protocol is used to produce stably transformed tobacco (Nicotiana tabacum) NT1 cell lines, using Agrobacterium tumefaciens-mediated DNA delivery of a binary vector containing a gene encoding hepatitis B surface antigen and a gene encoding the kanamycin selection marker. The NT1 cultures, at the appropriate stage of growth, are inoculated with A. tumefaciens containing the binary vector. A 3-day cocultivation period follows, after which the cultures are rinsed and placed on solid selective medium. Transformed colonies ('calli') appear in approximately 4 weeks; they are subcultured until adequate material is obtained for analysis of antigen production. 'Elite' lines are selected based on antigen expression and growth characteristics. The time required for the procedure from preparation of the plant cell materials to callus development is approximately 5 weeks. Growth of selected calli to sufficient quantities for antigen screening may require 4-6 weeks beyond the initial selection. Creation of the plasmid constructs, transformation of the A. tumefaciens line, and ELISA and Bradford assays to assess protein production require additional time. PMID- 17406391 TI - Sulforhodamine B colorimetric assay for cytotoxicity screening. AB - The sulforhodamine B (SRB) assay is used for cell density determination, based on the measurement of cellular protein content. The method described here has been optimized for the toxicity screening of compounds to adherent cells in a 96-well format. After an incubation period, cell monolayers are fixed with 10% (wt/vol) trichloroacetic acid and stained for 30 min, after which the excess dye is removed by washing repeatedly with 1% (vol/vol) acetic acid. The protein-bound dye is dissolved in 10 mM Tris base solution for OD determination at 510 nm using a microplate reader. The results are linear over a 20-fold range of cell numbers and the sensitivity is comparable to those of fluorometric methods. The method not only allows a large number of samples to be tested within a few days, but also requires only simple equipment and inexpensive reagents. The SRB assay is therefore an efficient and highly cost-effective method for screening. PMID- 17406392 TI - Assessing nest building in mice. AB - For small rodents, nests are important in heat conservation as well as reproduction and shelter. Nesting is easily measured in the home cages of mice, particularly with the advent of pressed cotton materials. The mice first shred the tightly packed material, then arrange it into a nest. Published studies have often used materials such as hay, twine or tissues, sometimes preshredded, and have assigned scores of the quality of the resulting nest with rather rudimentary rating scales; e.g., 0, no nest; 1, flat nest; 2, nest covering the mouse. The protocol described here uses pressed cotton squares and a definitive 5-point nest rating scale. Any unshredded material left after a bout of nesting can also be weighed, providing a semi-independent objective assay of nesting ability. Nesting has been shown to be sensitive to brain lesions, pharmacological agents and genetic mutations. This is a simple, cheap and easily done test that, along with other tests of species-typical behavior, is a sensitive assay for identifying previously unknown behavioral phenotypes. The test needs to be done overnight, but it should take no more than 5 minutes to set up plus 1 minute to assess one nest and weigh the untorn residue. PMID- 17406393 TI - Generation of peptide-MHC class I complexes through UV-mediated ligand exchange. AB - Major histocompatibility complex (MHC) class I molecules present peptide ligands on the cell surface for recognition by appropriate cytotoxic T cells. MHC-bound peptides are critical for the stability of the MHC complex, and standard strategies for the production of recombinant MHC complexes are based on in vitro refolding reactions with specific peptides. This strategy is not amenable to high throughput production of vast collections of MHC molecules. We have developed conditional MHC ligands that form stable complexes with MHC molecules but can be cleaved upon UV irradiation. The resulting empty, peptide-receptive MHC molecules can be charged with epitopes of choice under native conditions. Here we describe in-depth procedures for the high-throughput production of peptide-MHC (pMHC) complexes by MHC exchange, the analysis of peptide exchange efficiency by ELISA and the parallel production of MHC tetramers for T-cell detection. The production of the conditional pMHC complex by an in vitro refolding reaction can be achieved within 2 weeks, and the actual high-throughput MHC peptide exchange and subsequent MHC tetramer formation require less than a day. PMID- 17406394 TI - Transgenesis in fish: efficient selection of transgenic fish by co-injection with a fluorescent reporter construct. AB - Small fish are a popular laboratory model for studying gene expression and function by transgenesis. If, however, the transgenes are not readily detectable by visual inspection, a large number of embryos must be injected, raised and screened to identify positive founder fish. Here, we describe a strategy to efficiently generate and preselect transgenic lines harbouring any transgene of interest. Co-injection of a selectable reporter construct (e.g., GFP), together with the transgene of interest on a separate plasmid using the I-SceI meganuclease approach, results in co-distribution of the two plasmids. The quality of GFP expression within the F0 generation therefore reflects the quality of injection and allows efficient and reliable selection of founder fish that are also positive for the second transgene of interest. In our experience, a large fraction (up to 50%) of GFP-positive fish will also be transgenic for the second transgene, thus providing a rapid (within 3-4 months) and efficient way to establish transgenic lines for any gene of interest in medaka and zebrafish. PMID- 17406395 TI - Stromal cell-dependent growth of B-1 B cell progenitors in the absence of direct contact. AB - This protocol describes a transwell culture system in which stromal cells support the growth and differentiation of B cell progenitors in the absence of direct contact. In this system, a confluent layer of S17 stromal cells pre-established in 0.4 microm transwells is placed over wells seeded with purified B cell progenitors. The stromal cell-derived factors and additional cytokines added to the culture medium support the differentiation of the progenitors in the lower chamber. B-1 B cell progenitors seeded in the presence of thymic stromal lymphopoietin undergo significant expansion and differentiation in this culture system. Since the expanded B-1 B lineage cells are not contaminated with stromal cells, no additional purification steps are required before subsequent phenotypic, functional or genetic analyses of these lymphoid cells are performed. Once the transwell cultures and B cell progenitors are available, cultures can be initiated in less than an hour. The overall procedure, however, takes approximately 10 h when the initiation of the S17 transwell cultures and the isolation of the B cell progenitors steps are included. PMID- 17406396 TI - Production of chimeras by aggregation of embryonic stem cells with diploid or tetraploid mouse embryos. AB - The production of mouse chimeras is a common step in the establishment of genetically modified animal strains. Chimeras also provide a powerful experimental tool for following cell behavior during both prenatal and postnatal development. This protocol outlines a simple and economical technique for the production of large numbers of mouse chimeras using traditional diploid morula<- >diploid embryonic stem (ES) cell aggregations. Additional steps are included to describe the procedures necessary to produce specialized tetraploid chimeras using tetraploid morula<-->diploid ES cell aggregations. This increasingly popular form of chimera produces embryos of nearly complete ES cell derivation that can be used to speed transgenic production or ask developmental questions. Using this protocol, mouse chimeras can be generated and transferred to pseudopregnant surrogate mothers in a 5-d period. PMID- 17406397 TI - 220-plex microRNA expression profile of a single cell. AB - Here we describe a protocol for the detection of the microRNA (miRNA) expression profile of a single cell by stem-looped real-time PCR, which is specific to mature miRNAs. A single cell is first lysed by heat treatment without further purification. Then, 220 known miRNAs are reverse transcribed into corresponding cDNAs by stem-looped primers. This is followed by an initial PCR step to amplify the cDNAs and generate enough material to permit separate multiplex detection. The diluted initial PCR product is used as a template to check individual miRNA expression by real-time PCR. This sensitive technique permits miRNA expression profiling from a single cell, and allows analysis of a few cells from early embryos as well as individual cells (such as stem cells). It can also be used when only nanogram amounts of rare samples are available. The protocol can be completed in 7 d. PMID- 17406398 TI - Creating self-illuminating quantum dot conjugates. AB - Semiconductor quantum dots are inorganic fluorescent nanocrystals that, because of their unique optical properties compared with those of organic fluorophores, have become popular as fluorescent imaging probes. Although external light excitation is typically required for imaging with quantum dots, a new type of quantum dot conjugate has been reported that can luminesce with no need for external excitation. These self-illuminating quantum dot conjugates can be prepared by coupling of commercially available carboxylate-presenting quantum dots to the light-emitting protein Renilla luciferase. When the conjugates are exposed to the luciferase's substrate coelenterazine, the energy released by substrate catabolism is transferred to the quantum dots through bioluminescence resonance energy transfer, leading to quantum dot light emission. This protocol describes step-by-step procedures for the preparation and characterization of these self-illuminating quantum dot conjugates. The preparation process is relatively simple and can be done in less than 2 hours. The availability of self illuminating quantum dot conjugates will provide many new possibilities for in vivo imaging and detection, such as monitoring of in vivo cell trafficking, multiplex bioluminescence imaging and new quantum dot-based biosensors. PMID- 17406399 TI - Preparation of organotypic hippocampal slice cultures for long-term live imaging. AB - This protocol details a method to establish organotypic slice cultures from mouse hippocampus, which can be maintained for several months. The cultures are based on the interface method, which does not require special equipment, is easy to execute and yields slice cultures that can be imaged repeatedly--from when they are isolated at postnatal day 6-9, and up to 6 months in vitro. The preserved tissue architecture facilitates the analysis of defined hippocampal synapses, cells and entire projections. Monitoring of defined cellular and molecular components in the slices can be achieved by preparing slices from transgenic mice or by introducing transgenes through transfection or viral vectors. This protocol can be completed in 3 h. PMID- 17406400 TI - Multiplex-fluorescence in situ hybridization for chromosome karyotyping. AB - Multiplex-fluorescence in situ hybridization (M-FISH) was initially developed to stain human chromosomes--the 22 autosomes and X and Y sex chromosomes--with uniquely distinctive colors to facilitate karyotyping. The characteristic spectral signatures of all different combinations of fluorochromes are determined by multichannel image-analysis methods. Advantages of M-FISH include rapid analysis of metaphase spreads, even in complex cases with multiple chromosomal rearrangements, and identification of marker chromosomes. The M-FISH technology has been extended to other species, such as the mouse. Furthermore, in addition to painting probes, the method has been used with a variety of region-specific probes. M-FISH has even recently been used for 3D studies to analyze the distribution of human chromosomes in intact and preserved interphase nuclei. Hence, M-FISH has evolved into an essential tool for both clinical diagnostics and basic research. In this protocol, we describe how to use M-FISH to karyotype chromosomes, a procedure that takes approximately 14 d if new M-FISH probes have to be generated and 3 d if the M-FISH probes are ready to use. PMID- 17406401 TI - Determining the role of specific signaling molecules during lymphocyte development in vivo: instant transgenesis. AB - A common method of determining the role of specific signaling molecules during lymphocyte development is to generate a transgenic mouse. This procedure, while informative, is time consuming, expensive and ultimately does not guarantee a defined answer. Here we present a protocol in which the in vivo effects of a gene of interest on both B and T lymphocyte development may be determined simultaneously in a relatively short time period. This is achieved by introducing a defined gene, such as a wild-type or mutated signaling molecule, into a lymphoid progenitor population by retroviral infection. The retrovirus generates a bicistronic message encoding the gene of interest and GFP, thus enabling identification of retrovirally transduced cells in subsequent lymphocyte lineages. The cells are then introduced into mice deficient for recombinase activating gene 1 (Rag-/- mice), thus allowing the development of donor-derived B and T lymphocytes in vivo. Using this technique, results can be obtained within 3 8 weeks. PMID- 17406402 TI - Using drug-discrimination techniques to study the abuse-related effects of psychoactive drugs in rats. AB - Drug-discrimination (DD) techniques can be used to study abuse-related effects by establishing the interoceptive effects of a training drug (e.g., cocaine) as a cue for performing a specific operant response (e.g., lever pressing reinforced by food). During training with this protocol, pressing one lever is reinforced when the training drug is injected before the start of the session, and responding on a second lever is reinforced when vehicle is injected before the session. Lever choice during test sessions can then be used as an indication of whether a novel drug has effects similar to the training drug, or whether a potential therapeutic alters the effects of the training drug. Although training can be lengthy (up to several months), the pharmacological specificity of DD procedures make them a perfect complement to other techniques used to study drug abuse phenomena, such as intravenous self-administration and conditioned place preference procedures. PMID- 17406403 TI - A radioactive uptake assay to measure ion transport across ion channel-containing liposomes. AB - Here we describe a procedure for incorporating ion channels into lipid vesicles (liposomes) and functional characterization of the channel population by assaying radioactive isotope uptake into these proteoliposomes. The technique as described will work only for potassium channels but can be easily modified, as suggested in the text, for other ion channels and transporters. Purified ion channel proteins in detergent micelles are combined with solubilized lipids. Detergent is subsequently removed from protein-lipid complexes by gel filtration or dialysis into high potassium (high [K+]) buffer. After freezing-thawing and sonication, the resultant larger liposomes are passed over another gel-filtration column to exchange an extraliposomal high [K+] to a low [K+] buffer, thus establishing a large K+ gradient across the liposomal membrane. Trace 86Rb is then added to the extraliposomal space and the reaction begins. If the ion channel is permeable to K+, the K+ inside exits the liposomes down its concentration gradient and the 86Rb outside accumulates in the intraliposomal space until equilibrium is reached. The reaction time course is monitored by measurement of accumulated 86Rb after removal of external 86Rb over an ion-exchange column. The 86Rb flux assay takes 2-5 hours depending on the reaction rate and the number of desired time points. PMID- 17406404 TI - Expanded-bed adsorption immobilized-metal affinity chromatography. AB - The protocol describes a method for capture of secreted hexahistidine-tagged proteins using expanded-bed adsorption immobilized-metal affinity chromatography. The starting material for the procedure is any crude feedstock that contains a histidine (His)-tagged target protein. The protocol is exemplified using unclarified broth from Pichia pastoris fermentation as feedstock. The protocol can be used for laboratory studies or as part of a process for production of recombinant biotherapeutics to standards of good manufacturing practice. It takes approximately 5 h to purify proteins from 10 liters of feedstock and a further 5 6 h to sterilize and regenerate the column. PMID- 17406405 TI - Long-term live imaging of neuronal circuits in organotypic hippocampal slice cultures. AB - This protocol details a method for imaging organotypic slice cultures from the mouse hippocampus. The cultures are based on the interface method, which does not require special equipment, is easy to execute, and yields slice cultures that can be imaged repeatedly after they are isolated on postnatal day 6-9 and for up to 6 months in vitro. The preserved tissue architecture facilitates the analysis of defined hippocampal synapses, cells and entire projections. Time-lapse imaging is based on transgenes expressed in the mice, or on constructs introduced through transfection or viral vectors; it can reveal processes that develop over time periods ranging from seconds to months. Imaging can be repeated at least eight times without detectable morphological damage to neurons. Subsequent to imaging, the slices can be processed for immunocytochemistry or electron microscopy, to collect further information about the structures that have been imaged. This protocol can be completed in 35 min. PMID- 17406406 TI - Cerasome as an infusible and cell-friendly gene carrier: synthesis of cerasome forming lipids and transfection using cerasome. AB - Sonication of a pre-agitated aqueous solution of cationic lipid having a (EtO)3SiCH2CH2CH2 group on the quarternized ammonium nitrogen results in partially silica- or ceramic-coated liposome (cerasome), which can be used as an excellent transfection agent. Non-silylated reference lipid, which may represent cationic lipids that are used in conventional lipofection experiments, form a compact liposome, which undergoes DNA-induced fusion to provide transfection irrelevant and larger (100-300 nm), more toxic particles. The surface-rigidified cerasome is infusible and the monomeric cerasome complex of DNA is of viral size (approximately 70 nm) and exhibits a remarkable transfection performance with a 10(2)-10(3)-fold higher efficiency (relative to the non-silylated reference lipid), minimized cytotoxicity and serum compatibility. The cerasome lipid is obtained by the reaction of 3-bromopropyltriethoxysilane with a tertiary amine derivative of the lipid. Preparation of an aqueous cerasome solution takes 1-2 h. The cerasome-DNA complex and the transfection takes about 3 d to complete. PMID- 17406407 TI - Dendritic patch-clamp recording. AB - The patch-clamp technique allows investigation of the electrical excitability of neurons and the functional properties and densities of ion channels. Most patch clamp recordings from neurons have been made from the soma, the largest structure of individual neurons, while their dendrites, which form the majority of the surface area and receive most of the synaptic input, have been relatively neglected. This protocol describes techniques for recording from the dendrites of neurons in brain slices under direct visual control. Although the basic technique is similar to that used for somatic patching, we describe refinements and optimizations of slice quality, microscope optics, setup stability and electrode approach that are required for maximizing the success rate for dendritic recordings. Using this approach, all configurations of the patch-clamp technique (cell-attached, inside-out, whole-cell, outside-out and perforated patch) can be achieved, even for relatively distal dendrites, and simultaneous multiple electrode dendritic recordings are also possible. The protocol--from the beginning of slice preparation to the end of the first successful recording--can be completed in 3 h. PMID- 17406408 TI - Using phiC31 integrase to make transgenic Xenopus laevis embryos. AB - Bacteriophage phiC31 produces the enzyme integrase that allows the insertion of the phage genome into its bacterial host. This enzyme recognizes a specific DNA sequence in the phage (attP) and a different sequence in the bacterium (attB). Recombination between these sites leads to integration in a reaction that requires no accessory factors. Seminal studies by the Calos laboratory demonstrated that the phiC31 integrase was capable of integrating plasmid with an attB site into mammalian genomes at sites that approximated the attP site. We describe the use of attB-containing plasmids with insulated reporter genes for the successful integration of DNA into Xenopus embryos. The method offers a way to produce transgenic embryos without manipulation of sperm nuclei using microinjection methods that are standard for experiments in Xenopus laevis. The method aims to allow the non-mosaic controlled expression of new genetic material in the injected embryo and compares favorably with the time that is normally taken to analyze embryos injected with mRNAs, plasmids, morpholinos or oligonucleotides. PMID- 17406409 TI - Capping of CdSe-ZnS quantum dots with DHLA and subsequent conjugation with proteins. AB - We provide a detailed protocol for designing water-soluble CdSe-ZnS quantum dots (QDs) based on cap exchange of the native hydrophobic shell with dihydrolipoic acid (DHLA) ligands, and the preparation of functional QD bioconjugates for use in immunoassays. Our conjugation strategy is based on non-covalent self-assembly between DHLA-capped QDs and protein appended with either an electrostatic attachment domain (namely, the basic leucine zipper) or a polyhistidine tag. These bioconjugates combine the properties of the QD and attached biomolecule to create structures with desirable luminescent and biologically specific properties. This method also allows the preparation of mixed surface conjugates, which results in the conjugates gaining multiple biological activities. Conjugation of DHLA-capped QDs to maltose binding protein (MBP), the immunoglobulin-G-binding beta2 domain of streptococcal protein G (PG) and avidin will be described. MBP and PG were modified by genetic fusion with either a charged leucine zipper or a polyhistidine interaction domain. PMID- 17406410 TI - In vivo single-cell electroporation for transfer of DNA and macromolecules. AB - Single-cell electroporation allows transfection of plasmid DNA or macromolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and combinations thereof. Experienced users can electroporate roughly 40 tadpoles per hour. The technique can be adapted for use with other charged transfer materials and in other systems and tissues where cells can be targeted with a micropipette. Under visual guidance, an electrode filled with transfer material is placed in a cell body-rich area of the tadpole brain and a train of voltage pulses applied, which electroporates a nearby cell. We show examples of successfully electroporated single cells, instances of common problems and troubleshooting suggestions. Single-cell electroporation is an affordable method to fluorescently label and genetically manipulate individual cells. This powerful technique enables observation of single cells in an otherwise normal environment. PMID- 17406411 TI - Assaying RNA chaperone activity in vivo in bacteria using a ribozyme folding trap. AB - Here, we report an assay to evaluate the intracellular RNA chaperone activity of a protein of interest in vivo in bacterial cells. The method is based on self splicing of the group I intron, which is located in the thymidylate synthase (td) gene of phage T4. A previously described td mutant (tdSH1) has significantly impaired splicing due to formation of splicing-incompetent alternative structures. In this procedure, overexpression of RNA chaperones in the presence of the td mutant SH1 is used to evaluate whether the putative RNA chaperone is able to rescue the incorrectly folded group I intron. The ability of the RNA chaperone to assist during folding is measured indirectly by assessing the difference between the splicing efficiencies of the td mutant in the absence and in the presence of the RNA chaperone. This procedure can be completed in 5-6 d, not including the time needed to clone the putative RNA chaperone. PMID- 17406412 TI - Design and implementation of bimolecular fluorescence complementation (BiFC) assays for the visualization of protein interactions in living cells. AB - Bimolecular fluorescence complementation (BiFC) analysis enables direct visualization of protein interactions in living cells. The BiFC assay is based on the discoveries that two non-fluorescent fragments of a fluorescent protein can form a fluorescent complex and that the association of the fragments can be facilitated when they are fused to two proteins that interact with each other. BiFC must be confirmed by parallel analysis of proteins in which the interaction interface has been mutated. It is not necessary for the interaction partners to juxtapose the fragments within a specific distance of each other because they can associate when they are tethered to a complex with flexible linkers. It is also not necessary for the interaction partners to form a complex with a long half life or a high occupancy since the fragments can associate in a transient complex and un-associated fusion proteins do not interfere with detection of the complex. Many interactions can be visualized when the fusion proteins are expressed at levels comparable to their endogenous counterparts. The BiFC assay has been used for the visualization of interactions between many types of proteins in different subcellular locations and in different cell types and organisms. It is technically straightforward and can be performed using a regular fluorescence microscope and standard molecular biology and cell culture reagents. PMID- 17406413 TI - Immunocytochemistry and quantification of protein colocalization in cultured neurons. AB - This protocol details a method to quantify the distribution of protein and colocalization in neuronal cultures. To that end, this protocol includes itemized steps and considerations for performing immunocytochemistry, acquiring fluorescence images and quantifying multichannel fluorescence images. Success in quantifying immunostained neurons relies on the accessibility of the proteins of interest, the sensitivity and specificity of antibodies, the signal-to-noise ratio of the collected image and the sensitivity of the quantification method. In contrast to other commonly employed methods for quantification, the protocol detailed here requires manual selection of punctae and subtraction of background selected for each neurite. This approach reliably and uniquely allows for detection of proteins in low signal-to-noise ratio images, which are characteristic of developing neurons. Thus, this method serves an important niche in image analysis poorly addressed by alternative published methods. In general, immunocytochemistry requires 3.5-7 h, and one triple-immunostained neuron can be quantified in 1.5 h. PMID- 17406414 TI - Evaluating the biological relevance of putative enhancers using Tol2 transposon mediated transgenesis in zebrafish. AB - Evaluating the biological relevance of the myriad putative regulatory noncoding sequences in vertebrate genomes represents a huge challenge. Functional analyses in vivo have typically relied on costly and labor-intensive transgenic strategies in mice. Transgenesis has also been applied in nonrodent vertebrates, such as zebrafish, but until recently these efforts have been hampered by significant mosaicism and poor rates of germline transmission. We have developed a transgenic strategy in zebrafish based on the Tol2 transposon, a mobile element that was recently identified in another teleost, Medaka. This method takes advantage of the increased efficiency of genome integration that is afforded by this intact DNA transposon, activity that is mediated by the corresponding transposase protein. The approach described in this protocol uses a universal vector system that permits rapid incorporation of DNA that is tagged with sequence targets for site-specific recombination. To evaluate the regulatory potential of a candidate sequence, the desired interval is PCR-amplified using sequence-specific primers that are flanked by the requisite target sites for cloning, and recombined into a universal expression plasmid (pGW_cfosEGFP). Purified recombinant DNAs are then injected into 1-2-cell zebrafish embryos and the resulting reporter expression patterns are analyzed at desired timepoints during development. This system is amenable to large-scale application, facilitating rapid functional analysis of noncoding sequences from both mammalian and teleost species. PMID- 17406415 TI - Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study 'recognition memory'. AB - Rats and mice have a tendency to interact more with a novel object than with a familiar object. This tendency has been used by behavioral pharmacologists and neuroscientists to study learning and memory. A popular protocol for such research is the object-recognition task. Animals are first placed in an apparatus and allowed to explore an object. After a prescribed interval, the animal is returned to the apparatus, which now contains the familiar object and a novel object. Object recognition is distinguished by more time spent interacting with the novel object. Although the exact processes that underlie this 'recognition memory' requires further elucidation, this method has been used to study mutant mice, aging deficits, early developmental influences, nootropic manipulations, teratological drug exposure and novelty seeking. PMID- 17406416 TI - High efficiency germ-line transformation of mosquitoes. AB - The ability to manipulate the mosquito genome through germ-line transformation provides us with a powerful tool for investigating gene structure and function. It is also a valuable method for the development of novel approaches to combating the spread of mosquito-vectored diseases. To date, germ-line transformation has been demonstrated in several mosquito species. Transgenes are introduced into pre blastocyst mosquito embryos using microinjection techniques that take a few hours, and progeny are screened for the presence of a marker gene. The microinjection protocol presented here can be applied to most mosquitoes and contains several improvements over other published methods that increase the survival of injected embryos and, therefore, the number of transformants. Transgenic lines can be established in approximately 1 month using this technique. PMID- 17406417 TI - A general user interface for prediction servers of proteins' post-translational modification sites. AB - Post-translational modifications (PTMs) of proteins play essential roles in governing the functions and dynamics of proteins and are implicated in many cellular processes. Several types of PTMs have been investigated through computational approaches, including phosphorylation, sumoylation, palmitoylation, and lysine and arginine methylation, among others. Because the large diversity in the user interfaces (UIs) of different prediction servers for PTMs could possibly hinder experimental biologists in using these servers, we propose to develop a protocol for a unified UI for PTM prediction servers, based on our own work and that of other groups on PTM site prediction. By following this protocol, tool developers can provide a uniform UI regardless of the PTM types and the underlying computational algorithms. With such uniformity in the UI, experimental biologists would be able to use any PTM prediction server compliant with this protocol once they had learned to use one of them. It takes a typical PTM prediction server compliant with this unified UI several minutes to calculate the prediction results for a protein 1,000 amino acids in length. PMID- 17406418 TI - Microcontact printing of axon guidance molecules for generation of graded patterns. AB - Microcontact printing (microCP) of proteins has been successfully used for patterning surfaces in various contexts. Here we describe a simple 'lift-off' method to print precise patterns of axon guidance molecules, which are used as substrate for growing chick retinal ganglion cell (RGC) axons. Briefly, the etched pattern of a silicon master is transferred to a protein-coated silicone cuboid (made from polydimethylsiloxane, PDMS), which is then used as a stamp on a glass coverslip. RGC explants are placed adjacent to the pattern and cultured overnight. Fluorescent labeling of the printed proteins allows the quantitative analysis of the interaction of axons and growth cones with single protein dots and of the overall outgrowth and guidance rate in variously designed patterns. Patterned substrates can be produced in 3-4 h and are stable for up to one week at 4 degrees C; the entire protocol can be completed in 3 d. PMID- 17406419 TI - Design, construction and in vitro testing of zinc finger nucleases. AB - Zinc finger nucleases (ZFNs) are hybrid proteins that have been developed as targetable cleavage reagents for double-stranded DNA, both in vitro and in vivo. This protocol describes the design and construction of new DNA-binding domains comprised of zinc fingers (ZFs) directed at selected DNA sequences. Because the ZFNs must dimerize to cut DNA, they are designed in pairs for any new site. The first step is choosing a DNA segment of interest and searching it for sequences that can be recognized by combinations of existing ZFs. The second step is the construction of coding sequences for the selected ZF sets. Third, these coding sequences are linked to that of the nonspecific cleavage domain from the FokI restriction endonuclease in a cloning vector of choice. Finally, the ZFNs are expressed in Escherichia coli, partially purified, and tested in vitro for cleavage of the target sequences to which they were designed. If all goes smoothly, design, construction and cloning can be completed in about two weeks, with expression and testing completed in one additional week. PMID- 17406420 TI - Quantification of starch in plant tissues. AB - This protocol describes a simple means of measuring the starch content of plant tissues by solubilizing the starch, converting it quantitatively to glucose and assaying the glucose. Plant tissue must initially be frozen rapidly to stop metabolism, then extracted to remove free glucose. Starch is solubilized by heating, then digested to glucose by adding glucan hydrolases. Glucose is assayed enzymatically. The method is more sensitive and accurate than iodine-based protocols, and is suitable for tissues that have a wide range of starch contents. Measurements on multiple samples can be completed within a day. PMID- 17406422 TI - Two-dimensional difference gel electrophoresis. AB - Two-dimensional difference gel electrophoresis (2D DIGE) is a modified form of 2D electrophoresis (2DE) that allows one to compare two or three protein samples simultaneously on the same gel. The proteins in each sample are covalently tagged with different color fluorescent dyes that are designed to have no effect on the relative migration of proteins during electrophoresis. Proteins that are common to the samples appear as 'spots' with a fixed ratio of fluorescent signals, whereas proteins that differ between the samples have different fluorescence ratios. With the appropriate imaging system, DIGE is capable of reliably detecting as little as 0.5 fmol of protein, and protein differences down to +/- 15%, over a >10,000-fold protein concentration range. DIGE combined with digital image analysis therefore greatly improves the statistical assessment of proteome variation. Here we describe a protocol for conducting DIGE experiments, which takes 2-3 d to complete. PMID- 17406421 TI - Evolving proteins in mammalian cells using somatic hypermutation. AB - We describe a new method to mutate target genes through somatic hypermutation (SHM) and to evolve proteins directly in living mammalian cells. Target genes are expressed under the control of an inducible promoter in a B-cell line that hypermutates its immunoglobulin (Ig) V genes constitutively. Mutations can be introduced into the target gene through SHM upon transcription. Mutant genes are then expressed and selected or screened for desired properties in cells. Identified cells are subjected to another round of mutation and selection or screening. This process can be iterated easily for numerous rounds, and multiple reinforcing mutations can be accumulated to produce desirable phenotypes. This approach bypasses labor-intensive in vitro mutagenesis and samples a large protein sequence space. In this protocol a monomeric red fluorescent protein (mRFP1.2) was evolved in Ramos cells to afford a mutant (mPlum) with far-red emission. This method can be adapted to evolve other eukaryotic proteins and to be used in other cells able to perform SHM. For each round of evolution, it takes approximately 1 d to mutate the target gene, approximately 0.5-1 d to select or screen, and 2-4 d to propagate the cells for the next round depending on how many cells are collected. PMID- 17406423 TI - Non-SELEX: selection of aptamers without intermediate amplification of candidate oligonucleotides. AB - Aptamers are typically selected from libraries of random DNA (or RNA) sequences through systematic evolution of ligands by exponential enrichment (SELEX), which involves several rounds of alternating steps of partitioning of candidate oligonucleotides and their PCR amplification. Here we describe a protocol for non SELEX selection of aptamers--a process that involves repetitive steps of partitioning with no amplification between them. Non-equilibrium capillary electrophoresis of equilibrium mixtures (NECEEM), which is a highly efficient affinity method, is used for partitioning. NECEEM also facilitates monitoring of bulk affinity of enriched libraries at every step of partitioning and screening of individual clones for their affinity to the target. NECEEM allows all clones to be screened prior to sequencing, so that only clones with suitable binding parameters are sequenced. The entire protocol can be completed in 1 wk, whereas conventional SELEX protocols take several weeks even in a specialized industrial facility. PMID- 17406424 TI - Developing multiplexed SNP assays with special reference to degraded DNA templates. AB - This protocol describes a single nucleotide polymorphism (SNP) genotyping strategy for highly degraded DNA, using a two-stage multiplex whereby multiple fragments are first amplified in a single exponential reaction and the products of this PCR are added to a linear single-base-extension reaction. It utilizes the analytical power of a capillary electrophoresis system to simultaneously type all the target sites. The protocol is specifically written for use with severely fragmented templates, typical of ancient DNA, and can be adapted to widely used detection platforms. The addition of the single-phase genotyping step avoids the need for the re-amplification and cloning of PCR products, while providing its own controls for the detection of contamination and allelic drop-out. This protocol can facilitate the routine analysis of up to 52 SNP markers (haploid or diploid) in 96 samples in a single day, and is recommended for the authentication of data in all areas of DNA research (population and medical genetics, forensics, ancient DNA). PMID- 17406425 TI - Micropatterned cell cultures on elastic membranes as an in vitro model of myocardium. AB - We describe here a new in vitro protocol for structuring cardiac cell cultures to mimic important aspects of the in vivo ventricular myocardial phenotype by controlling the location and mechanical environment of cultured cells. Microlithography is used to engineer microstructured silicon metal wafers. Those are used to fabricate either microgrooved silicone membranes or silicone molds for microfluidic application of extracellular matrix proteins onto elastic membranes (involving flow control at micrometer resolution). The physically or microfluidically structured membranes serve as a cell culture growth substrate that supports cell alignment and allows the application of stretch. The latter is achieved with a stretching device that can deliver isotropic or anisotropic stretch. Neonatal ventricular cardiomyocytes, grown on these micropatterned membranes, develop an in vivo-like morphology with regular sarcomeric patterns. The entire process from fabrication of the micropatterned silicon metal wafers to casting of silicone molds, microfluidic patterning and cell isolation and seeding takes approximately 7 days. PMID- 17406426 TI - Using tRNA-linked molecular beacons to image cytoplasmic mRNAs in live cells. AB - Imaging products of gene expression in live cells will provide unique insights into the biology of cells. Molecular beacons make attractive probes for imaging mRNA in live cells as they can report the presence of an RNA target by turning on the fluorescence of a quenched fluorophore. However, when oligonucleotide probes are introduced into cells, they are rapidly sequestered in the nucleus, making the detection of cytoplasmic mRNAs difficult. We have shown that if a molecular beacon is linked to a tRNA, it stays in the cytoplasm and permits detection of cytoplasmic mRNAs. Here we describe two methods of linking molecular beacons to tRNA and show how the joint molecules can be used for imaging an mRNA that is normally present in the cytoplasm in live cultured cells. This protocol should take a total of 4 d to complete. PMID- 17406427 TI - BrdU assay for neurogenesis in rodents. AB - Neurogenesis within the adult central nervous system is demonstrated using an exogenous cell tracer, 5'-bromo-2'-deoxyuridine (BrdU), in combination with endogenous neuronal markers. Specific primary antibodies raised against these markers are widely available and their visualization is possible with the use of fluorescently tagged secondary antibodies. BrdU is a thymidine analog that incorporates into dividing cells during DNA synthesis. Once incorporated into the new DNA, BrdU will remain in place and be passed down to daughter cells following division. Typically, BrdU is injected intraperitoneally. Different survival times required by the desired experimental time-line will yield data on specific phases of neurogenesis: proliferation, differentiation and maturation. One of the drawbacks of using BrdU is the dependence on a stressful injection procedure and uncertain penetration of the targeted cells with a uniform concentration of the compound. Thus, for experiments requiring measurements of cell proliferation, Ki67 can be used as an acceptable alternative. The protocol takes 3-5 d, allowing for sectioning and staining. PMID- 17406428 TI - Methylation-specific digital karyotyping. AB - Epigenetic alterations, including DNA methylation and histone modifications, are known to regulate various physiological and pathological processes. In mammalian cells, DNA methylation occurs at cytosines of CpG dinucleotides. Several methods have been developed for the genome-wide analysis of methylation patterns. However, none of these are quantitative or sequence-based, and the identification of the exact location of the methylated CpG is difficult. In this protocol, we describe a recently developed method--methylation-specific digital karyotyping (MSDK)--that enables comprehensive and unbiased genome-wide DNA methylation analysis. Using a combination of a methylation-sensitive mapping enzyme (for example, AscI) and a fragmenting enzyme (for example, NlaIII), short sequence tags can be obtained and uniquely mapped to genome location. The number of tags in an MSDK library reflects the methylation status of the mapping enzyme sites. Generation of MSDK libraries can be completed in 7-10 days, whereas sequencing and data analysis requires an additional 3-4 weeks. PMID- 17406429 TI - Generation of transgenic birds with replication-deficient lentiviruses. AB - Birds are of great interest as an animal model in biological research and for commercial applications as a bioreactor. Effective methods for manipulating the avian genome would accelerate progress in fields such as developmental biology and behavioral neurobiology, which traditionally have relied on birds as model systems for biological research. Here we describe a simple and effective protocol for producing transgenic birds using lentiviral vectors that can be used to achieve tissue-specific transgene expression at high levels. The time allotted for the procedure depends upon the species of bird; adult transgenic quails can be generated in approximately 5 months. PMID- 17406430 TI - Production and characterization of adeno-associated viral vectors. AB - The adeno-associated virus (AAV) is one of the most promising viral vectors for human gene therapy. As with any potential therapeutic system, a thorough understanding of it at the in vitro and in vivo levels is required. Over the years, numerous methods have been developed to better characterize AAV vectors. These methods have paved the way to a better understanding of the vector and, ultimately, its use in clinical applications. This review provides an up-to-date, detailed description of essential methods such as production, purification and titering and their application to characterize current AAV vectors for preclinical and clinical use. PMID- 17406431 TI - Whole-body imaging with fluorescent proteins. AB - The intrinsic brightness of fluorescent proteins has been taken advantage of to develop a technology of whole-body imaging of tumors and gene expression in mouse internal organs. Stable transformation with fluorescent protein genes can be effected using retroviral vectors containing a selectable marker such as neomycin resistance. The cells that stably express fluorescent proteins can then be transplanted into appropriate mouse models. For whole-body imaging, nude mice are very appropriate. If wild-type mice are used, then hair must be removed by shaving or depilation. The instruments used can range from a simple LED flashlight and appropriate excitation and emission filters to sophisticated equipment such as the Olympus OV100 with a wide range of magnification, enabling both macroimaging and microimaging. It is crucial that proper filters be used such that background autofluorescence is minimal. Fluorescent protein-based imaging technology can be used for whole-body imaging of fluorescent cells on essentially all organs. The timeline for these experiments varies from 2 days to 2 months. PMID- 17406432 TI - Preparation of organotypic hippocampal slice cultures: interface method. AB - This protocol describes a method for making and culturing rat hippocampal organotypic slices on membrane inserts. Supplementary videos are included to demonstrate visually the different steps of the procedure. Cultured hippocampal slices has been increasingly used as a model for synaptic studies of the brain as they allow examination of mid to long term manipulations in a preparation where the gross cytoarchitecture of the hippocampus is preserved. Combining techniques such as molecular biology, electrophysiology and immunohistochemistry to study physiological or pathological processes can easily be applied to organotypic slices. The technique described here can be used to make organotypic slices from other parts of the brain, other rodent species and from a range of ages. This protocol can be completed in 3 h. PMID- 17406433 TI - Protein profiling by the combination of two independent mass spectrometry techniques. AB - Protein profiling in the high-throughput mode is a most useful technique that allows formation of reference databases for cells and tissues and performance of comparative proteomics. In the proposed protocol protein extraction from tissues is followed by 2D gel electrophoresis (2DE) with subsequent in-gel digestion and identification of soluble proteins by two individual mass spectrometric techniques, tandem matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and nano-liquid chromatography (nano-LC)-MS/MS. The proposed combined use of these two MS approaches leads to a very high identification rate of well separated protein spots from a gel. In the first step 2DE separates high abundance proteins (those visualized by nonsensitive Coomassie blue staining) that are subsequently picked, digested and aliquoted for MS applications. Protein samples not identified by MALDI-MS or MS/MS (77% of all spots) are finally unambiguously identified by nano-LC-MS/MS (total identification rate 94%). This protocol can be completed in 6 weeks. PMID- 17406434 TI - Carbon fiber technique for the investigation of single-cell mechanics in intact cardiac myocytes. AB - This protocol describes a method for attaching single isolated cardiac myocytes to carbon fibers for mechanical manipulation and measurement. This method relies on cell-adhesive carbon fibers that attach easily to the cell membrane without causing damage, and is thus applicable to intact myocytes. To connect the carbon fiber to micromanipulators, a fiber holder with glass capillaries must first be fabricated. After connection of the fibers to the micromanipulators, firm attachment is easily established by gently pressing the fiber tip onto the cell membrane. Unlike other methods, this technique does not require vast technical expertise, and therefore greatly facilitates experiments. This method enables detection of the effect of drugs, genetic defects or the expression of exogenous proteins on both active and passive properties of cardiac myocytes. In combination with other experimental procedures, this technique can also be applied to the study of mechano-transduction. This protocol can be completed in 3.5 h. PMID- 17406435 TI - Analysis of apoptosis by propidium iodide staining and flow cytometry. AB - Since its introduction, the propidium iodide (PI) flow cytometric assay has been widely used for the evaluation of apoptosis in different experimental models. It is based on the principle that apoptotic cells, among other typical features, are characterized by DNA fragmentation and, consequently, loss of nuclear DNA content. Use of a fluorochrome, such as PI, that is capable of binding and labeling DNA makes it possible to obtain a rapid (the protocol can be completed in about 2 h) and precise evaluation of cellular DNA content by flow cytometric analysis, and subsequent identification of hypodiploid cells. The original protocol enhanced the capacity for a rapid, quantitative measure of cell apoptosis. For this reason, since its publication, the PI assay has been widely used, as demonstrated by the large number of citations of the original paper and/or the continuous use of the method in many laboratories. PMID- 17406436 TI - In situ hybridization for mRNA detection in Arabidopsis tissue sections. AB - Plant biology is currently confronted with an overflow of expression profile data provided by high-throughput microarray transcription analyses. However, the tissue and cellular resolution of these techniques is limited. Thus, it is still necessary to examine the expression pattern of selected candidate genes at a cellular level. Here we present an in situ mRNA hybridization method that is routinely used in the analysis of gene expression patterns. The protocol is optimized for mRNA localizations in sectioned tissue of Arabidopsis seedlings including embryos, roots, hypocotyls, young primary leaves and flowers. The detailed protocol, recommended controls and troubleshooting are presented along with examples of application. The total time for the process is 10 days. PMID- 17406437 TI - The lineaging of fluorescently-labeled Caenorhabditis elegans embryos with StarryNite and AceTree. AB - Lineage analysis of Caenorhabditis elegans is a powerful tool for characterizing developmental phenotypes and embryonic gene-expression patterns. We present a detailed protocol for the lineaging of embryos by computational analysis of 4D images of embryos that ubiquitously express histone-GFP (green fluorescent protein) fusion proteins through the 350 cell stage followed by manual editing. We describe how to optimize imaging settings for this purpose, the use of the lineage-extraction software, StarryNite, and the lineage-editing software, AceTree. In addition, we describe a useful polymer bead mounting technique for C. elegans embryos that has several advantages compared with the standard agar pad mounting technique. The protocol requires about 1 h of user time spread over 2 days to generate the raw lineage, and an additional 2 or 4 h to edit the lineage to the 194- or 350-cell stage, respectively. PMID- 17406438 TI - Methods for MAG3 conjugation and 99mTc radiolabeling of biomolecules. AB - The chelator mercaptoacetyltriglycine (MAG3) forms a single stable chelate with technetium-99m (99mTc) oxotechnetate. The bifunctional N-hydroxysuccinimidyl ester of mercaptoacetyltriglycine with S-acetyl protection of the sulfhydryl group may be used to conjugate MAG3 to primary amine functionalized biomolecules for the purpose of radiolabeling with 99mTc for gamma detection or single photon emission computed tomography imaging (SPECT). We report here an improved MAG3 conjugation and 99mTc radiolabeling method capable of generating high radiochemical yield and high specific radioactivity. Post-labeling purification will not be needed if the protocol is followed as presented. Apart from the preparation of reagents, the conjugation and purification requires about 4 h, while the labeling with 99mTc requires about an additional 30 min. PMID- 17406439 TI - Serial analysis of binding elements for human transcription factors. AB - Serial analysis of binding elements (SABE) is a method that can be used to identify the genome-wide location of transcription factor binding sites in human or other mammalian cells. In this method, a specific antibody targeting a DNA binding transcription factor of interest is used to pull down the transcription factor and its bound DNA elements through chromatin immunoprecipitation (ChIP). ChIP DNA fragments are further enriched by subtractive hybridization against non enriched DNA using representational difference analysis (RDA) and analyzed through the generation of sequence tags similar to serial analysis of gene expression (SAGE). The SABE method circumvents the need for microarrays and is able to identify immunoprecipitated loci in an unbiased manner. The combination of ChIP, RDA and SAGE-type methods has advantages over other similar strategies in reducing the level of intrinsic noise sequences that are typically present in ChIP samples from human cells. This protocol takes about 2 weeks to complete. PMID- 17406440 TI - Tissue-specific and cell type-specific RNA interference in vivo. AB - RNA interference (RNAi) is an efficient method for silencing genes in cultured cells. Here we describe a simple RNAi approach for silencing genes in a cell type specific and tissue-specific way in vivo. The approach, which mimics the means by which naturally occurring 'microRNA's are generated, uses a tissue-specific polymerase II promoter to drive the expression of a short hairpin RNA (shRNA) directed against the gene target. The shRNA is cleaved by ubiquitously expressed endonucleases to form an active small interfering RNA of about 22 nt. As a proof of principle, it has been shown that expression of a shRNA directed against the transcription factor Wilms tumor 1 in transgenic mice reduces that protein specifically in nurse cells in the testis. Our transgenic RNAi approach offers a cost-effective means of rapidly (within months) addressing the function(s) of genes of interest in a wide variety of specific cell types and tissues in mice in vivo. PMID- 17406441 TI - An ex vivo method for rapid generation of monoclonal antibodies (ADLib system). AB - Here, we describe a protocol for using the ADLib (Autonomously Diversifying Library) system to rapidly generate specific monoclonal antibodies using DT40, a chicken B-cell line that undergoes constitutive gene conversion at both light- and heavy-chain immunoglobulin loci. We previously developed the ADLib system on the basis of our finding that gene conversion in DT40 cells was enhanced by treatment of the cells with a histone deacetylase inhibitor, trichostatin A (TSA). TSA treatment evolves a diversified library of DT40 cells (ADLib), in which each cell has different surface IgM specificity. Antigen-specific DT40 cells are selected from ADLib using antigen-conjugated magnetic beads, and their specificity can be examined by various immunological assays, using culture supernatant containing secreted IgM. The whole process from selection to screening can be completed in about 1 week. Thus, the ADLib system will accelerate biological studies, including drug discovery and design. PMID- 17406442 TI - Intracellular cytokine optimization and standard operating procedure. AB - We describe here a method for optimizing the use of polychromatic flow cytometry (with up to 17 fluorochromes simultaneously) in surface and intracellular staining of human T lymphocytes. We will highlight and discuss how to procedurally optimize key steps in the experimental process before an intracellular cytokine staining assay protocol is finalized. These include but are not limited to the titration of monoclonal antibodies, use of a dead-cell discriminator and 'dump' channel, selection of a cytokine secretion inhibitor, selection of fixation and permeabilization reagents, and inclusion of compensation controls. Building on this basic protocol, we then establish a polychromatic assay designed to detect five separate functions of T lymphocytes (production of three cytokines and one chemokine, and degranulation) while simultaneously identifying multiple surface markers on the responding cells. PMID- 17406443 TI - Diolistic labeling of neuronal cultures and intact tissue using a hand-held gene gun. AB - Diolistic labeling is a highly efficient method for introducing dyes into cells using biolistic techniques. The use of lipophilic carbocyanine dyes, combined with particle-mediated biolistic delivery using a hand-held gene gun, allows non toxic labeling of multiple cells in both living and fixed tissue. The technique is rapid (labeled cells can be visualized in minutes) and technically undemanding. Here, we provide a detailed protocol for diolistic labeling of cultured human embryonic kidney 293 cells and whole brain using a hand-held gene gun. There are four major steps: (i) coating gold microcarriers with one or more dyes; (ii) transferring the microcarriers into a cartridge to make a bullet; (iii) preparation of cells or intact tissue; and (iv) firing the microcarriers into cells or tissue. The method can be readily adapted to other cell types and tissues. This protocol can be completed in less than 1 h. PMID- 17406444 TI - Quality assurance for polychromatic flow cytometry. AB - This protocol outlines a three-part quality assurance program to optimize, calibrate and monitor flow cytometers used to measure cells labeled with five or more fluorochromes (a practice known as polychromatic flow cytometry). The initial steps of this program (system optimization) ensure that the instrument's lasers, mirrors and filters are optimally configured for the generation and transmission of multiple fluorescent signals. To determine the sensitivity and dynamic range of each fluorescence detector, the system is then calibrated by measuring fluorescence over a range of photomultiplier tube (PMT) voltages by determining the PMT voltage range and linearity (Steps 2-10) and validating the PMT voltage (Steps 11-17). Finally, to ensure consistent performance, we provide procedures to monitor the precision, accuracy and sensitivity of fluorescence measurements over time. All three aspects of this program should be performed upon installation, or whenever changes occur along the flow cytometer's optical path. However, only a few of these procedures need to be carried out on a routine basis. PMID- 17406445 TI - Analysis of translation initiation using a translation control reporter system. AB - The study of translational control has become increasingly important, as aberrant translation has been linked to the etiology of human diseases. Nevertheless, a convenient research tool to measure and quantify cellular translational activity has not been developed to date. Here we present a translation control reporter system (TCRS) for straightforward and accurate analysis of cellular translational activity. Our method relies on the expression of two unique reporter peptides from a single messenger RNA transcript. Using TCRS-expressing cell lines, changes in initiation of translation have been detected in response to translationally active drugs. Accordingly, TCRS may promote the discovery of novel agents that modulate translation. TCRS may also be used in the identification of signal transduction pathways that impinge on translation control. Furthermore, the modular design allows the exchange of regulatory cassettes for the examination of other putative cis-regulatory mRNA elements. The time required for the procedure depends on whether transient TCRS expression is used or stable TCRS-expressing cell lines have to be produced and will range from 5 to 14 d, respectively. PMID- 17406446 TI - Two-step metal affinity purification of double-tagged (NusA-His6) fusion proteins. AB - The present purification protocol applies to target proteins that are fused to a double tag, such as NusA-His6, through a linker that includes a protease recognition sequence. It involves two steps of immobilized metal ion affinity chromatography (IMAC). NusA stabilizes the passenger protein during translation, whereas the His-tag enables affinity purification of the fusion. The eluate resulting from the first IMAC is buffer-exchanged to remove the imidazole and to achieve optimal conditions for the enzymatic cleavage performed by a His-tagged recombinant protease. The digested sample is loaded directly for a second IMAC step and the target protein is selectively recovered in the flow-through. The resin binds residual non-digested fusion protein, double-tagged moiety, protease and any contaminant that bound the affinity resin and was eluted from the first IMAC. The purity of the target protein usually makes a further purification step unnecessary for most of the lab applications. It takes less than 5 hours to purify the protein from a 5 g pellet. PMID- 17406447 TI - Negative detection of biomolecules separated in polyacrylamide electrophoresis gels. AB - Here we describe the protocols for negative or reverse detection of proteins, nucleic acids and lipopolysaccharides separated in polyacrylamide electrophoresis gels. These protocols are based on the selective synthesis and precipitation of a white imidazole-zinc complex in the gel, which is absent from those zones where biomolecules are located. These methods are highly sensitive (1-10 ng of biomolecules per band), very cheap as they use inexpensive, common laboratory reagents (imidazole and a Zn II salt), rapid (less than 20 min after gel washing), robust and simple (two steps). Reverse-stained biomolecules are reversibly fixed in the gel. After brief incubation in a zinc chelating agent, biomolecules can be recovered from the gel with the same efficiency as from unstained gels. In consequence, they are procedures of choice for micropreparative applications. References covering typical applications are included. PMID- 17406448 TI - In vivo electroporation in the embryonic mouse central nervous system. AB - This protocol describes a basic method for in vivo electroporation in the nervous system of embryonic mice. Delivery of electric pulses following microinjection of DNA into the brain ventricle or the spinal cord central canal enables efficient transfection of genes into the nervous system. Transfection is facilitated by forceps-type electrodes, which hold the uterus and/or the yolk sac containing the embryo. More than ten embryos in a single pregnant mouse can be operated on within 30 min. More than 90% of operated embryos survive and more than 90% of these survivors express the transfected genes appropriately. Gene expression in neurons persists for a long time, even at postnatal stages, after electroporation. Thus, this method could be used to analyze roles of genes not only in embryonic development but also in higher order function of the nervous system, such as learning. PMID- 17406449 TI - Quantification of mRNA using real-time RT-PCR. AB - The real-time reverse transcription polymerase chain reaction (RT-qPCR) addresses the evident requirement for quantitative data analysis in molecular medicine, biotechnology, microbiology and diagnostics and has become the method of choice for the quantification of mRNA. Although it is often described as a "gold" standard, it is far from being a standard assay. The significant problems caused by variability of RNA templates, assay designs and protocols, as well as inappropriate data normalization and inconsistent data analysis, are widely known but also widely disregarded. As a first step towards standardization, we describe a series of RT-qPCR protocols that illustrate the essential technical steps required to generate quantitative data that are reliable and reproducible. We would like to emphasize, however, that RT-qPCR data constitute only a snapshot of information regarding the quantity of a given transcript in a cell or tissue. Any assessment of the biological consequences of variable mRNA levels must include additional information regarding regulatory RNAs, protein levels and protein activity. The entire protocol described here, encompassing all stages from initial assay design to reliable qPCR data analysis, requires approximately 15 h. PMID- 17406450 TI - Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol. AB - The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay has been modified for real-time, quantitative PCR analysis. Here, we describe cost-effective procedures for detection of telomerase activity using a fluorescent-based assay as well as by using real-time PCR. These modified TRAP assays can be accomplished within 4 h (from lysis of samples to analysis of telomerase products). PMID- 17406451 TI - Quantification of cerebral amyloid angiopathy and parenchymal amyloid plaques with Congo red histochemical stain. AB - In the current protocol, we describe the Congo red staining method and a method for separately quantifying vascular and parenchymal amyloid deposits in brain tissue sections. Congo red staining detects amyloid deposits in brain tissue of amyloid precursor protein transgenic mice and human Alzheimer's tissue. It detects compacted amyloid in a beta-sheet secondary structure and labels amyloid in both the brain parenchyma (amyloid plaques) and blood vessels. Congophilic amyloid in blood vessels is called cerebral amyloid angiopathy (CAA). To date, analysis of CAA has largely used a severity rating scale, including both qualitative and quantitative characteristics. Here, we describe a simple method for quantifying total Congophilic staining and resolution of this staining into the parenchymal and vascular components based on morphological criteria. It is becoming increasingly important to separately quantify various components of the Alzheimer's pathology, given the advancement of amyloid-lowering therapies into clinical trials. The entire procedure for the Congo red staining can be performed at room temperature (20-25 degrees C) in a fume hood. The staining protocol should take 1 h 30 min including time for coverslipping slides. Time required for image analysis depends greatly on the number of samples being analyzed and the software being used. In our hands, 30 images can be collected per hour and quantified in a further 2 h. PMID- 17406452 TI - A rodent model of myocardial infarction for testing the efficacy of cells and polymers for myocardial reconstruction. AB - We have developed a robust rat model of myocardial infarction (MI). Here we describe the step-by-step protocol for creating an ischemia-reperfusion rat model of MI. We also describe how to deliver therapeutic injections of mesenchymal stem cells (MSCs) together with fibrin, to show an application of this model. In addition, to confirm the presence of fibrin and cells in the infarct, visualization of MSCs and fibrin by histological techniques are also described. The ischemia-reperfusion MI model can be modified and generalized for use with various injectable polymers, cell types, drugs, DNA and combinations thereof. The model can be created in 7 days or less, depending on the timing of therapeutic intervention. PMID- 17406453 TI - Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE): quantitative RNA structure analysis at single nucleotide resolution. AB - Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) interrogates local backbone flexibility in RNA at single-nucleotide resolution under diverse solution environments. Flexible RNA nucleotides preferentially sample local conformations that enhance the nucleophilic reactivity of 2'-hydroxyl groups toward electrophiles, such as N-methylisatoic anhydride (NMIA). Modified sites are detected as stops in an optimized primer extension reaction, followed by electrophoretic fragment separation. SHAPE chemistry scores local nucleotide flexibility at all four ribonucleotides in a single experiment and discriminates between base-paired versus unconstrained or flexible residues with a dynamic range of 20-fold or greater. Quantitative SHAPE reactivity information can be used to establish the secondary structure of an RNA, to improve the accuracy of structure prediction algorithms, to monitor structural differences between related RNAs or a single RNA in different states, and to detect ligand binding sites. SHAPE chemistry rarely needs significant optimization and requires two days to complete for an RNA of 100-200 nucleotides. PMID- 17406454 TI - RNAi screens in Caenorhabditis elegans in a 96-well liquid format and their application to the systematic identification of genetic interactions. AB - We describe a protocol for performing RNA interference (RNAi) screens in Caenorhabditis elegans in liquid culture in 96-well plates. The procedure allows a single researcher to set-up and score RNAi experiments at approximately 2,000 genes per day. By comparing RNAi phenotypes between wild-type worms and worms carrying a defined genetic mutation, we have used this protocol to identify synthetic lethal interactions between genes systematically. We also describe how the protocol can be adapted to target two genes simultaneously by combinatorial RNAi. PMID- 17406455 TI - Standardized reagents and protocols for engineering zinc finger nucleases by modular assembly. AB - Engineered zinc finger nucleases can stimulate gene targeting at specific genomic loci in insect, plant and human cells. Although several platforms for constructing artificial zinc finger arrays using "modular assembly" have been described, standardized reagents and protocols that permit rapid, cross-platform "mixing-and-matching" of the various zinc finger modules are not available. Here we describe a comprehensive, publicly available archive of plasmids encoding more than 140 well-characterized zinc finger modules together with complementary web based software (termed ZiFiT) for identifying potential zinc finger target sites in a gene of interest. Our reagents have been standardized on a single platform, enabling facile mixing-and-matching of modules and transfer of assembled arrays to expression vectors without the need for specialized knowledge of zinc finger sequences or complicated oligonucleotide design. We also describe a bacterial cell-based reporter assay for rapidly screening the DNA-binding activities of assembled multi-finger arrays. This protocol can be completed in approximately 24 26 d. PMID- 17406456 TI - An isothermal primer extension method for whole genome amplification of fresh and degraded DNA: applications in comparative genomic hybridization, genotyping and mutation screening. AB - We describe a protocol that uses a bioinformatically optimized primer in an isothermal whole genome amplification (WGA) reaction. Overnight incubation at 37 degrees C efficiently generates several hundred- to several thousand-fold increases in input DNA. The amplified product retains reasonably faithful quantitative representation of unamplified whole genomic DNA (gDNA). We provide protocols for applying this isothermal primer extension WGA protocol in three different techniques of genomic analysis: comparative genomic hybridization (CGH), genotyping at simple tandem repeat (STR) loci and screening for single base mutations in a common monogenic disorder, beta-thalassemia. gDNA extracted from formalin-fixed paraffin-embedded (FFPE) tissues can also be amplified with this protocol. PMID- 17406457 TI - Xenopus laevis transgenesis by sperm nuclear injection. AB - The stable integration of transgenes into embryos of the frog Xenopus laevis is achieved using the procedure described here. Linear DNA containing the transgene is incorporated randomly into sperm nuclei that have had their membranes disrupted with detergent treatment. Microinjection of these nuclei into unfertilized eggs produces viable embryos that can be screened for activity of the transgene. The proportion of embryos that harbor the transgene varies from 10 to 40% of the total number of surviving embryos. Multiple copies of the transgene can integrate as a concatemer into the sperm genome, and more than one site of DNA integration might occur within resulting animals. Germ cell transmission of the transgene is routine and the procedure is well suited to the production of transgenic reporter frog lines. One day should be allocated for the preparation of the sperm nuclei, which are stored as aliquots for future use. The transgenesis reaction and egg injection take one morning. PMID- 17406458 TI - Genomic systematic evolution of ligands by exponential enrichment (Genomic SELEX) for the identification of protein-binding RNAs independent of their expression levels. AB - Genomic systematic evolution of ligands by exponential enrichment (Genomic SELEX) is an experimental procedure for the expression condition-independent identification of protein-binding RNAs. RNA libraries derived from genomic DNA are generated via random priming, PCR amplification and in vitro transcription. Libraries consist of genomic sequences of selected size, and fragments are flanked by constant sequences required for amplification and transcription. This RNA pool is then subjected to several rounds of selection and amplification to enrich for RNAs meeting the selection criteria. Various selection criteria are possible. Here we describe selection by affinity to a protein of interest. High affinity ligands can then be cloned and sequenced to allow their identification. With this method, protein-binding RNAs can be discovered, nucleic acid-protein interactions can be identified, and whole protein-nucleic acid networks can be defined. This method is also suitable for discovering novel genes, including non protein-coding RNAs, and it complements in silico approaches. It is better suited to detect protein-binding RNAs that are differentially expressed (and therefore absent from many tissues) and low-abundance RNAs than experimental procedures that start from the isolation of expressed RNAs. The protocol takes approximately 3 months to complete. PMID- 17406460 TI - Fluorometric measurement of nitrite/nitrate by 2,3-diaminonaphthalene. AB - We describe a step-by-step protocol for measuring the stable products of the nitric oxide (NO) pathway: nitrite, nitrite plus nitrate and nitrate. This described protocol is easy to apply and is about 50 times more sensitive than the commonly used Griess reaction or commercially available assay kits based on the Griess reaction. It also allows the study of minimal changes in the NO pathway. With this method, it takes about 3 h to analyze the above-mentioned stable products in culture supernatants or in various body fluids, and the method has a sensitive linear range of 0.02-10.0 microM. This restricted linear range suggests that the technique is useful for studying small changes of nitrite and nitrate, rather than for routine diagnostic measurements. PMID- 17406459 TI - Verification of automated peptide identifications from proteomic tandem mass spectra. AB - Shotgun proteomics yields tandem mass spectra of peptides that can be identified by database search algorithms. When only a few observed peptides suggest the presence of a protein, establishing the accuracy of the peptide identifications is necessary for accepting or rejecting the protein identification. In this protocol, we describe the properties of peptide identifications that can differentiate legitimately identified peptides from spurious ones. The chemistry of fragmentation, as embodied in the 'mobile proton' and 'pathways in competition' models, informs the process of confirming or rejecting each spectral match. Examples of ion-trap and tandem time-of-flight (TOF/TOF) mass spectra illustrate these principles of fragmentation. PMID- 17406461 TI - A rapid method for chloroplast isolation from the green alga Chlamydomonas reinhardtii. AB - This method has been developed to yield highly purified intact chloroplasts from Chlamydomonas reinhardtii. This procedure involves breaking cell-wall-deficient cells by passage through a narrow-bore syringe needle and purifying the intact chloroplasts by differential centrifugation and Percoll gradient centrifugation. This procedure can be completed in less than 3 h and is capable of generating relatively high yields of chloroplasts that should be useful for researchers studying the biochemistry and cell biology of C. reinhardtii chloroplasts. PMID- 17406462 TI - Isolation of Caenorhabditis elegans gene knockouts by PCR screening of chemically mutagenized libraries. AB - This protocol details methodologies to generate Caenorhabditis elegans deletion mutants by chemical mutagenesis and to detect them by PCR screening. Approximately, 600,000 worms are grown synchronously, mutagenized with ethyl methane sulfonate, divided in groups of 500 and allowed to self-fertilize for two generations. DNA is prepared from a fraction of each worm population, pooled into a 96-well plate, and screened by PCR with primers positioned 2.5-3.5 kb apart. Cultures containing deletion mutants are subdivided in small worm populations and tested again by PCR to identify positives. Single animals are then cloned from positive cultures, allowed to self-fertilize and identified by PCR genotyping. This method, which takes about a month, gives approximately a 50% chance of finding a deletion of interest larger than 500-600 bp. If a deletion cannot be found, the library can be pooled at lower complexity and screened for smaller deletions using an alternative PCR-based method. PMID- 17406463 TI - PEGylation of native disulfide bonds in proteins. AB - PEGylation has turned proteins into important new biopharmaceuticals. The fundamental problems with the existing approaches to PEGylation are inefficient conjugation and the formation of heterogeneous mixtures. This is because poly(ethylene glycol) (PEG) is usually conjugated to nucleophilic amine residues. Our PEGylation protocol solves these problems by exploiting the chemical reactivity of both of the sulfur atoms in the disulfide bond of many biologically relevant proteins. An accessible disulfide bond is mildly reduced to liberate the two cysteine sulfur atoms without disturbing the protein's tertiary structure. Site-specific PEGylation is achieved with a bis-thiol alkylating PEG reagent that sequentially undergoes conjugation to form a three-carbon bridge. The two sulfur atoms are re-linked with PEG selectively conjugated to the bridge. PEGylation of a protein can be completed in 24 h and purification of the PEG-protein conjugate in another 3 h. We have successfully applied this approach to PEGylation of cytokines, enzymes, antibody fragments and peptides, without destroying their tertiary structure or abolishing their biological activity. PMID- 17406464 TI - Identification of fungal cell wall mutants using susceptibility assays based on Calcofluor white and Congo red. AB - The fungal cell wall is an essential organelle and represents a considerable metabolic investment. Its macromolecular composition, molecular organization and thickness can vary greatly depending on environmental conditions. Its construction is also tightly controlled in space and time. Many genes are therefore involved in building the fungal cell wall. Here we present a simple approach for detecting these genes. The method is based on the observation that cell wall mutants are generally more sensitive to two related anionic dyes, Calcofluor white (CFW) and Congo red (CR), both of which interfere with the construction and stress response of the cell wall. CFW-based and CR-based susceptibility assays identify cell wall mutants not only in ascomycetous yeasts (such as Saccharomyces cerevisiae and Candida albicans) but also in mycelial ascomycetes (such as Aspergillus fumigatus and Aspergillus niger), basidiomycetous species (Cryptococcus neoformans) and probably also zygomycetous fungi. The protocol can be completed in 4-6 h (excluding the incubation time required for fungal growth). PMID- 17406465 TI - Recombinant protein purification by self-cleaving aggregation tag. AB - A simple technique is presented for non-chromatographic purification of recombinant proteins expressed in Escherichia coli. This method is based on a reversibly precipitating, self-cleaving purification tag. The tag is made up of two components: an elastin-like polypeptide (ELP), which reversibly self associates in high-salt buffers at temperatures above 30 degrees C; and an intein, which causes the ELP tag to self-cleave in response to a mild pH shift. Thus, a tripartite ELP-intein-target protein precursor can be purified by cycles of salt addition, heating and centrifugation. Once purified, intein-mediated self cleavage, followed by precipitation of the cleaved ELP tag, allows easy and effective isolation of the pure, native target protein without the need for chromatographic separations. Recoveries of 50-100 mg of cleaved, native target protein per liter of shake-flask culture have been achieved for over a dozen proteins, typically in 8-24 h depending on specific process parameters. PMID- 17406466 TI - Collection of lymph-borne dendritic cells in the rat. AB - Dendritic cells (DCs) are crucial in immune induction. Not only do they collect antigens in peripheral tissues, and transport and process them for presentation to lymphocytes in draining lymph nodes, but they also regulate the immune response by modulating T-cell differentiation. Intestinal and hepatic DCs migrating in lymph can be collected from rats under near-physiological conditions. Initially, the mesenteric or celiac lymph nodes are removed from young rats (30 min). The afferent and efferent lymph vessels subsequently heal, permitting DCs to enter the thoracic duct. After at least 6 wk, the duct is cannulated (40 min). Lymph can be collected for up to 48 h. DCs can subsequently be identified, enriched and sorted to high degrees of purity. This two-stage technique generates large numbers of immunologically relevant DCs under near physiological conditions. Lymph collection requires 2-3 h per animal over 6 wk. PMID- 17406467 TI - Estimation of baculovirus titer based on viable cell size. AB - In this paper, a simple and rapid protocol for determination of baculovirus titers based on increasing viable insect cell size/diameter following virus infection is presented. There are different methods available for determining virus titers such as plaque assays end-point dilution, quantitative real-time polymerase chain reaction and flow cytometry. However, most of these methods are time consuming and labor intensive. The titer estimation method presented here can be completed in approximately 28 h from start to finish. In this method, the Vi-CELL (Beckman Coulter) was used to measure cell diameter change over a range of virus dilutions, following infection. The cell diameter change data were used to compute the virus titer using a statistical method called the method of moments that we have described previously. PMID- 17406468 TI - Administration and interpretation of the Trail Making Test. AB - Measurement of cognitive functions is an increasingly important goal for clinicians and researchers. Many neuropsychological test batteries are comprehensive and require specialized training to administer and interpret. The Trail Making Test is an accessible neuropsychological instrument that provides the examiner with information on a wide range of cognitive skills and can be completed in 5-10 min. Its background, psychometric properties, administration procedures and interpretive guidelines are provided in this protocol. PMID- 17406469 TI - Determination of plant resistance to carbamate herbicidal compounds inhibiting cell division and early growth by seed and plantlets bioassays. AB - Herbicide-resistant plants can be generated by either traditional breeding procedures or genetic engineering. Analyses of plant responses to a newly developed herbicide or the tolerance level of a newly developed plant line to a given herbicide are based on various bioassays. Here, we describe several methods for quantitative measurements of plants' responses to propham application, as a model herbicide of the carbamate family. Dose-response assays include seed germination and analyses of shoot and root elongation on paper. To better reflect the natural interaction between the plant, the soil and the herbicide, a protocol for germination and root elongation on sand is described. Finally, a more sensitive bioassay is based on plant growth on agar medium. The described protocols are simple, reproducible and can be easily adopted for a variety of plant species and for various herbicides. Plants' response to a given herbicide can be determined within a few weeks. PMID- 17406470 TI - Micropatterning of living cells by laser-guided direct writing: application to fabrication of hepatic-endothelial sinusoid-like structures. AB - Here, we describe a simple protocol for the design and construction of a laser guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells "on the fly" with micrometer precision on arbitrary matrices, including soft gels such as Matrigel. By micropatterning endothelial cells on Matrigel, one can control the self-assembly of vascular structures and associated liver tissue. LGDW is therefore uniquely suited for studying the role of tissue architecture and mechanical properties at the single-cell resolution, and for studying the effects of heterotypic cell-cell interactions underlying processes such as liver morphogenesis, differentiation and angiogenesis. The total time required to carry out this protocol is typically 7 h. PMID- 17406471 TI - Chemical cleavage of mismatch (CCM) to locate base mismatches in heteroduplex DNA. AB - This protocol describes the use of the chemical cleavage of mismatch (CCM) method to assess whether a region of DNA contains mutations and to localize them. Compared with other mutation-detection techniques (such as single strand conformation polymorphism (SSCP) analysis, denaturing high-performance liquid chromatography (DHPLC) and denaturing gradient gel electrophoresis (DGGE)) that detect mutations in short DNA fragments and require highly specific melting temperatures, CCM has a higher diagnostic sensitivity suited to the detection of mutations in tumor genes, and can analyze amplicons < or = 2 kb in length. To detect mutations, PCR heteroduplexes are incubated with two mismatch-specific reagents. Hydroxylamine modifies unpaired cytosine and potassium permanganate modifies unpaired thymine. The samples are then incubated with piperidine, which cleaves the DNA backbone at the site of the modified mismatched base. Cleavage products are separated by electrophoresis, revealing the identity and location of the mutation. The CCM method can efficiently detect point mutations as well as insertions and deletions. This protocol can be completed in 10 h. PMID- 17406472 TI - Investigating mitotic spindle assembly and function in vitro using Xenopus laevis egg extracts. AB - Extracts from Xenopus laevis eggs provide a powerful system for the study of cell division processes in vitro through biochemical reconstitution and manipulation, and microscopic analysis. We provide protocols for the preparation of metaphase arrested extracts and in vitro assays to examine the following pathways of spindle assembly: 1) Sperm nuclei added to meiotic extracts, supporting the formation of half-spindles and bipolar spindle structures around unreplicated chromosomes; 2) sperm nuclei added to extracts that cycle through interphase and form spindles that are capable of undergoing anaphase and chromosome segregation; and 3) spindle formation around chromatin-coated beads. Finally, we describe methods to inhibit a specific protein by immunodepletion or addition of an inhibitor such as a dominant-negative construct. These techniques can be used to analyze the mitotic function of a given protein. It takes approximately 1.5 h to prepare the extract, 1-3 h for spindle-assembly experiments and an additional 1-3 h if immunodepletion is performed. PMID- 17406473 TI - Clonogenic assay of cells in vitro. AB - Clonogenic assay or colony formation assay is an in vitro cell survival assay based on the ability of a single cell to grow into a colony. The colony is defined to consist of at least 50 cells. The assay essentially tests every cell in the population for its ability to undergo "unlimited" division. Clonogenic assay is the method of choice to determine cell reproductive death after treatment with ionizing radiation, but can also be used to determine the effectiveness of other cytotoxic agents. Only a fraction of seeded cells retains the capacity to produce colonies. Before or after treatment, cells are seeded out in appropriate dilutions to form colonies in 1-3 weeks. Colonies are fixed with glutaraldehyde (6.0% v/v), stained with crystal violet (0.5% w/v) and counted using a stereomicroscope. A method for the analysis of radiation dose-survival curves is included. PMID- 17406474 TI - A modified protocol for rapid DNA isolation from plant tissues using cetyltrimethylammonium bromide. AB - We describe a modification of the DNA extraction method, in which cetyltrimethylammonium bromide (CTAB) is used to extract nucleic acids from plant tissues. In contrast to the original method, the modified CTAB procedure is faster, omits the selective precipitation and CsCl gradient steps, uses less expensive and toxic reagents, requires only inexpensive laboratory equipment and is more readily adapted to high-throughput DNA extraction. This protocol yields approximately 5-30 microg of total DNA from 200 mg of tissue fresh weight, depending on plant species and tissue source. It can be completed in as little as 5-6 h. PMID- 17406475 TI - Cloning strategy, production and purification of proteins with exopeptidase cleavable His-tags. AB - Here, we present a cloning strategy for the production of recombinant proteins tagged with a polyhistidine sequence that can be cleaved by the exopeptidase, DAPase. The method can be used with most commonly available vectors and results in the expression of a His-tag protein that can be purified in its native form regardless of its natural sequence. This approach takes advantage of the TAGZyme system for the removal of amino-terminal affinity tags. Tag removal is accomplished either with DAPase (a recombinant dipeptidyl peptidase) alone or in combination with two accessory enzymes, Qcyclase and pGAPase. The system has been used for the production of intracellular proteins in Escherichia coli and can be applied to other expression hosts for the production of secreted proteins or proteins that require post-translational modification. The production of human interleukin 1beta in E. coli is used as an example to illustrate this method. The complete protocol from initial PCR to the production of a detagged protein with its authentic N terminus can be performed within 5 days. PMID- 17406476 TI - Cell culture and infection system for hepatitis C virus. AB - Hepatitis C virus (HCV) infection causes chronic liver disease and is a worldwide health problem. Despite ever-increasing demand for knowledge on viral replication and pathogenesis, detailed analysis has been hampered by a lack of efficient viral culture systems. We isolated HCV genotype 2a strain JFH-1 from a patient with fulminant hepatitis. This strain replicates efficiently in Huh7 cells. Efficient replication and secretion of recombinant viral particles can be obtained in cell culture by transfection of in vitro-transcribed full-length JFH 1 RNA into Huh7 cells. JFH-1 virus generated in cell culture is infectious for both naive Huh7 cells and chimpanzees. The efficiency of viral production and infectivity of generated virus is substantially improved with permissive cell lines. This protocol describes how to use this system, which provides a powerful tool for studying viral life cycle and for the construction of antiviral strategies and the development of effective vaccines. Viral particles can be obtained in 12 days with this protocol. PMID- 17406477 TI - High-throughput screening of activity and enantioselectivity of esterases. AB - A procedure for the high-throughput screening of esterases is described. This includes enzyme expression in microtiter plates and the measurement of activity and enantioselectivity (E) of the esterase variants using acetates of secondary alcohols as model substrates. Acetic acid released is converted in an enzyme cascade leading to the stoichiometric formation of NADH, which is quantified in a spectrophotometer. The method allows screening of several thousand mutants per day and has already been successfully applied to identify an esterase mutant with an E>100 toward an important building block for organic synthesis. This protocol can also be used for lipases and possibly other hydrolases that are expressed in soluble form in conventional Escherichia coli strains. This protocol can be completed in 3-4 days. PMID- 17406478 TI - A method for the covalent capture and screening of diverse small molecules in a microarray format. AB - This protocol describes a robust method for the covalent capture of small molecules with diverse reactive functional groups in microarray format, and outlines a procedure for probing small-molecule microarrays (SMMs) with proteins of interest. A vapor-catalyzed, isocyanate-mediated surface immobilization scheme is used to attach bioactive small molecules, natural products and small molecules derived from diversity-oriented synthesis pathways. Additionally, an optimized methodology for screening SMMs with purified proteins and cellular lysates is described. Finally, a suggested model for data analysis that is compatible with commercially available software is provided. These procedures enable a platform capability for discovering novel interactions with potential applications to immunoglobulin profiling, comparative analysis of cellular states and ligand discovery. With the appropriate materials and experimental setup, the printing of SMMs can be completed in 14 hours over 3 days. Screening and data analysis requires 2 days. A detailed timeline is provided. PMID- 17406479 TI - DNA methylation: bisulphite modification and analysis. AB - DNA methylation is an important epigenetic modification of DNA in mammalian genomes. DNA methylation patterns are established early in development, modulated during tissue-specific differentiation and disrupted in many disease states, including cancer. To understand further the biological functions of these changes, accurate and reproducible methods are required to fully analyze the DNA methylation sequence. Here, we describe the 'gold-standard' bisulphite conversion protocol that can be used to re-sequence DNA from mammalian cells in order to determine and quantify the methylation state of a gene or genomic region at single-nucleotide resolution. The process of bisulphite treatment exploits the different sensitivities of cytosine and 5-methylcytosine (5-MeC) to deamination by bisulphite under acidic conditions--in which cytosine undergoes conversion to uracil, whereas 5-MeC remains unreactive. Bisulphite conversion of DNA, in either single tubes or in a 96-well format, can be performed in a minimum of 8 h and a maximum of 18 h, depending on the amount and quality of starting DNA. PMID- 17406480 TI - Flow cytometry and FISH to measure the average length of telomeres (flow FISH). AB - Telomeres have emerged as crucial cellular elements in aging and various diseases including cancer. To measure the average length of telomere repeats in cells, we describe our protocols that use fluorescent in situ hybridization (FISH) with labeled peptide nucleic acid (PNA) probes specific for telomere repeats in combination with fluorescence measurements by flow cytometry (flow FISH). Flow FISH analysis can be performed using commercially available flow cytometers, and has the unique advantage over other methods for measuring telomere length of providing multi-parameter information on the length of telomere repeats in thousands of individual cells. The accuracy and reproducibility of the measurements is augmented by the automation of most pipetting (aspiration and dispensing) steps, and by including an internal standard (control cells) with a known telomere length in every tube. The basic protocol for the analysis of nucleated blood cells from 22 different individuals takes about 12 h spread over 2-3 days. PMID- 17406481 TI - Synthesis of non-natural ManNAc analogs for the expression of thiols on cell surface sialic acids. AB - The sialic acid biosynthetic pathway in mammalian cells utilizes N-acetyl-D mannosamine (ManNAc) as a natural metabolic precursor and has the remarkable ability to biosynthetically process non-natural ManNAc analogs. Herein, we describe a recipe-style protocol for the synthesis of the novel peracetylated analog Ac5ManNTGc (1) that contains a pendant acetylthio- group and enables incorporation of thiol functionalities into the glycocalyx of living cells. We also describe the synthesis of the oxygen analog Ac5ManNGc (2), which serves as an appropriate control compound for biological experiments with 1. Both 1 and 2 were prepared from a reported, common intermediate 8, which is selectively acetylated at the hydroxyl groups. In contrast to previous methods, this synthetic approach introduces O-acetyl groups first, followed by N-acylation. Starting from the commercially available D-mannosamine hydrochloride (5), gram quantities of both 1 and 2 can be prepared over five steps in about 2-3 weeks. PMID- 17406482 TI - Strategies for site-specific protein biotinylation using in vitro, in vivo and cell-free systems: toward functional protein arrays. AB - This protocol details methodologies for the site-specific biotinylation of proteins using in vitro, in vivo and cell-free systems for the purpose of fabricating functional protein arrays. Biotinylation of recombinant proteins, in vitro as well as in vivo, relies on the chemoselective reaction between cysteine biotin and a reactive thioester group at the C-terminus of a protein generated via intein-mediated cleavage. The cell-free system utilizes low concentrations of biotin-conjugated puromycin. Unlike other approaches that require tedious and costly downstream steps of protein purification, C-terminal biotinylated proteins can be captured directly onto avidin-functionalized slides from a mixture of other cellular proteins to generate the corresponding protein array. These methods were designed to maintain the integrity and activity of proteins in a microarray format, which potentially allows simultaneous functional assays of thousands of proteins. Assuming that the target proteins have been cloned into the expression vector, transformation of bacterial strain and growth of starter culture would take approximately 2 days. Expression and in vitro protein purification and biotinylation will take approximately 3 days whereas the in vivo method would take approximately 2 days. The cell-free protein biotinylation strategy requires only 6-8 h. PMID- 17406483 TI - Time-lapse imaging of dendritic spines in vitro. AB - Dendritic spines are small protrusions present postsynaptically at approximately 90% of excitatory synapses in the brain. Spines undergo rapid spontaneous changes in shape that are thought to be important for alterations in synaptic connectivity underlying learning and memory. Visualization of these dynamic changes in spine morphology are especially challenging because of the small size of spines (approximately 1 microm). Here we describe a microscope system, based on a spinning-disk confocal microscope, suitable for imaging mature dendritic spines in brain slice preparations, with a time resolution of seconds. We discuss two commonly used in vitro brain slice preparations and methods for transfecting them. Preparation and transfection require approximately 1 d, after which slices must be cultured for at least 21 d to obtain spines of mature morphology. We also describe imaging and computer analysis routines for studying spine motility. These procedures require in the order of 2 to 4 h. PMID- 17406484 TI - Culturing hippocampal neurons. AB - We provide protocols for preparing low-density dissociated-cell cultures of hippocampal neurons from embryonic rats or mice. The neurons are cultured on polylysine-treated coverslips, which are suspended above an astrocyte feeder layer and maintained in serum-free medium. When cultured according to this protocol, hippocampal neurons become appropriately polarized, develop extensive axonal and dendritic arbors and form numerous, functional synaptic connections with one another. Hippocampal cultures have been used widely for visualizing the subcellular localization of endogenous or expressed proteins, for imaging protein trafficking and for defining the molecular mechanisms underlying the development of neuronal polarity, dendritic growth and synapse formation. Preparation of glial feeder cultures must begin 2 weeks in advance, and it takes 5 d to prepare coverslips as a substrate for neuronal growth. Dissecting the hippocampus and plating hippocampal neurons takes 2-3 h. PMID- 17406485 TI - Extraction and analysis of soluble inositol polyphosphates from yeast. AB - Soluble inositol polyphosphates are implicated in the regulation of many important cellular functions. This protocol to extract and separate inositol polyphosphates from Saccharomyces cerevisiae is divided into three steps: labeling of yeast, extraction of soluble inositol polyphosphates and chromatographic separation. Yeast cells are incubated with tritiated inositol, which is taken up and metabolized into different phosphorylated forms. Soluble inositol polyphosphates are then acid-extracted and fractionated by high performance liquid chromatography. The radioactivity of each fraction is determined by scintillation counting. This highly sensitive and reproducible method allows the accurate detection of subtle changes in the inositol polyphosphate profile and takes less than 48 h. It can easily be applied to other systems and we have included two adaptations of the protocol, one optimized for mammalian cells and the other for Arabidopsis thaliana. PMID- 17406486 TI - A fluorescence-based protocol for quantifying angiotensin-converting enzyme activity. AB - The determination of angiotensin-converting enzyme (ACE) activity represents a useful tool in the study of different health pathologies, such as hypertension. This protocol describes a fluorescent assay for measuring ACE activity in vitro with high precision and sensitivity. The method relies on the ability of ACE to hydrolyse the internally quenched fluorescent substrate o-aminobenzoylglycyl-p nitro-L-phenylalanyl-L-proline. The generation of the fluorescent product o aminobenzoylglycine can be continuously monitored, preferably using a microtiter plate fluorometer, though the use of a conventional cuvette fluorometer would also be possible. The method has important advantages with respect to other assays, because it involves only a one-step reagent, is easy to carry out and allows the analysis of an elevated number of samples in shorter times. It can be completed in one and a half hours. In addition, the fact that all reagents are commercially available allows the rapid introduction of the assay into the laboratory. PMID- 17406487 TI - Multiplex-terminal restriction fragment length polymorphism. AB - A novel method called "multiplex-terminal restriction fragment length polymorphism (M-TRFLP)" has been recently developed which can be used for simultaneous analysis of the community composition of two or more microbial taxa (up to four). This method can also be used for microbial diagnostic purposes. For M-TRFLP analysis, primers specific to different target genes are used for multiplex-PCR, with one primer for each target being labeled with a unique fluorescent dye at its 5' end. Restriction digestion of the amplified products followed by fragment size analysis on a DNA sequencer produces profiles for targeted genes, which can be distinguished from each other by the color of the terminal fragments imparted by the unique fluorescent dye used for primer labeling. In contrast to current protocols, M-TRFLP allows multiple communities or multiple targets (genes) data to be obtained in just one reaction and therefore saves time, cost and labor. This protocol can be completed in 5-8 h. PMID- 17406488 TI - Triflimide (HNTf2)-catalyzed aldehyde cross-aldol reaction using "super silyl" enol ethers. AB - The synthesis of the acetaldehyde-derived tris(trimethylsilyl)silyl (super silyl) enol ether is described, as well as its use in the high-yielding aldehyde cross aldol reaction. The super silyl enol ether shows unprecedented reactivity in giving the 1:1 adduct in very high yield. This reaction is catalyzed by 0.05 mol% of the Bronsted acid triflimide (HNTf2) and is complete within 15 min, making the protocol very attractive for large-scale synthesis. PMID- 17406489 TI - Identification of eukaryotic secreted and cell surface proteins using the yeast secretion trap screen. AB - Secreted and cell surface proteins play essential roles in numerous essential biological processes in eukaryotic organisms, but are often more difficult to isolate and identify than proteins that are localized in intracellular compartments. However, several high-throughput 'gene-trap' techniques have been developed to characterize these 'secretomes', including the yeast secretion trap (YST) screen. This method involves fusing cDNA libraries from the tissue or cell type of interest to a yeast (Saccharomyces cerevisiae) invertase reporter gene, transforming the resulting fusion library into an invertase-deficient yeast strain and plating the transformants on a medium containing sucrose as the sole carbon source. A yeast cell with a transgene encoding a secreted or cell surface protein can synthesize a secreted invertase fusion protein that can rescue the mutant, and the plasmid DNA can then be sequenced to identify the gene that encodes it. We describe a recently improved version of this screen, which allows the identification of genes encoding secreted proteins in 1-2 months. PMID- 17406490 TI - Measuring the optokinetic response of zebrafish larvae. AB - Our laboratory screens for visual mutants by examining larval eye movements in response to rotating illuminated stripes. This behavior, which is termed an optokinetic response (OKR), is a reflex that appears in zebrafish at the same time as the development of the visual system. The OKR can be accurately measured by 4 d post-fertilization, which is the age when larvae begin foraging for food. The OKR requires approximately 1 min per larva analyzed. After identifying fish with defective eye movements, we conduct secondary screens (such as histological analysis and electroretinography) to identify the subset of fish with disruptions in the function of the outer retina. This paper describes our protocol for the OKR. Our setup is simple to construct and the materials needed are inexpensive. This makes our system especially useful for new undergraduate and graduate students, as well as introductory science lecturers. PMID- 17406491 TI - Staining protocol for organotypic hippocampal slice cultures. AB - This protocol details a method to immunostain organotypic slice cultures from mouse hippocampus. The cultures are based on the interface method, which does not require special equipment, is easy to execute and yields slice cultures that can be imaged repeatedly, from the time of isolation at postnatal day 6-9 up to 6 months in vitro. The preserved tissue architecture facilitates the analysis of defined hippocampal synapses, cells and entire projections. Time-lapse imaging is based on transgenes expressed in the mice or on constructs introduced through transfection or viral vectors; it can reveal processes that develop over periods ranging from seconds to months. Subsequent to imaging, the slices can be processed for immunocytochemistry to collect further information about the imaged structures. This protocol can be completed in 3 d. PMID- 17406492 TI - High-throughput knockout screen in fission yeast. AB - We have designed the most efficient strategy to knock out genes in fission yeast Schizosaccharomyces pombe on a large scale. Our technique is based on knockout constructs that contain regions homologous to the target gene cloned into vectors carrying dominant drug-resistance markers. Most of the steps are carried out in a 96-well format, allowing simultaneous deletion of 96 genes in one batch. Based on our knockout technique, we designed a strategy for cloning knockout constructs for all predicted fission yeast genes, which is available in a form of a searchable database http://mendel.imp.ac.at/Pombe_deletion/. We validated this technique in a screen where we identified novel genes required for chromosome segregation during meiosis. Here, we present our protocol with detailed instructions. Using this protocol, one person can knock out 96 S. pombe genes in 8 days. PMID- 17406493 TI - A protocol for TILLING and Ecotilling in plants and animals. AB - We describe Targeting-Induced Local Lesions IN Genomes (TILLING), a reverse genetic strategy for the discovery and mapping of induced mutations. TILLING is suitable for essentially any organism that can be mutagenized. The TILLING procedure has also been adapted for the discovery and cataloguing of natural polymorphisms, a method called Ecotilling. To discover nucleotide changes within a particular gene, PCR is performed with gene-specific primers that are end labeled with fluorescent molecules. After PCR, samples are denatured and annealed to form heteroduplexes between polymorphic DNA strands. Mismatched base pairs in these heteroduplexes are cleaved by digestion with a single-strand specific nuclease. The resulting products are size-fractionated using denaturing polyacrylamide gel electrophoresis and visualized by fluorescence detection. The migration of cleaved products indicates the approximate location of nucleotide polymorphisms. Throughput is increased and costs are reduced by sample pooling, multi-well liquid handling and automated gel band mapping. Once genomic DNA samples have been obtained, pooled and arrayed, thousands of samples can be screened daily. PMID- 17406494 TI - IRAP and REMAP for retrotransposon-based genotyping and fingerprinting. AB - Retrotransposons can be used as markers because their integration creates new joints between genomic DNA and their conserved ends. To detect polymorphisms for retrotransposon insertion, marker systems generally rely on PCR amplification between these ends and some component of flanking genomic DNA. We have developed two methods, retrotransposon-microsatellite amplified polymorphism (REMAP) analysis and inter-retrotransposon amplified polymorphism (IRAP) analysis, that require neither restriction enzyme digestion nor ligation to generate the marker bands. The IRAP products are generated from two nearby retrotransposons using outward-facing primers. In REMAP, amplification between retrotransposons proximal to simple sequence repeats (microsatellites) produces the marker bands. Here, we describe protocols for the IRAP and REMAP techniques, including methods for PCR amplification with a single primer or with two primers and for agarose gel electrophoresis of the product using optimal electrophoresis buffers and conditions. This protocol can be completed in 1-2 d. PMID- 17406496 TI - Error-prone rolling circle amplification: the simplest random mutagenesis protocol. AB - A simple protocol to introduce random mutations, named error-prone rolling circle amplification (RCA), is described. A template plasmid is amplified by RCA in the presence of MnCl2 and used for transformation of a host strain to give a mutant library with three to four random point mutations per kilobase throughout the entire plasmid. The prime advantage of this method is its simplicity. This protocol requires neither the design of specific primers nor the exploration of thermal cycling conditions. It takes just 10 min to prepare the reaction mixture, followed by overnight incubation and transformation of a host strain. This method permits rapid preparation of randomly mutated plasmid libraries, and will enable the wider adoption of random mutagenesis. PMID- 17406495 TI - One-pot three-enzyme chemoenzymatic approach to the synthesis of sialosides containing natural and non-natural functionalities. AB - Chemoenzymatic synthesis, which combines the flexibility of chemical synthesis and the high selectivity of enzymatic synthesis, is a powerful approach to obtain complex carbohydrates. It is a preferred method for synthesizing sialic acid containing structures, including those with diverse naturally occurring and non natural sialic acid forms, different sialyl linkages and different glycans that link to the sialic acid. Starting from N-acetylmannosamine, mannose or their chemically or enzymatically modified derivatives, sialic acid aldolase-catalyzed condensation reaction leads to the formation of sialic acids and their derivatives. These compounds are subsequently activated by a CMP-sialic acid synthetase and transferred to a wide range of suitable acceptors by a suitable sialyltransferase for the formation of sialosides containing natural and non natural functionalities. The three-enzyme coupled synthesis of sialosides can be carried out in one pot without the isolation of intermediates. The time for synthesis is 4-18 h. Purification and characterization of the product can be completed within 2-3 d. PMID- 17406497 TI - A peroxidase-coupled continuous absorbance plate-reader assay for flavin monoamine oxidases, copper-containing amine oxidases and related enzymes. AB - This absorbance plate-reader-based assay is suitable for the examination of monoamine oxidase and copper amine oxidase activities versus numerous substrates. The assay is robust, continuous, rapid, highly quantitative, reasonably sensitive, inexpensive and suitable for automation. In the presence of a suitable amine substrate, amine oxidase enzymes generate hydrogen peroxide, which then drives the peroxidase-dependent oxidation of 4-aminoantipyrine. A subsequent interaction with vanillic acid generates stoichiometric amounts of a red quinoneimine dye, the appearance of which is monitored at 498 nm. An alternative procedure in which vanillic acid is replaced by 2,4-dichlorophenol enhances sensitivity but precludes the measurement of monoamine oxidases due to inhibition of these enzymes by dichlorophenol. Some substrates with low redox potentials, such as catecholamines, are not suitable for inclusion in this assay. A researcher familiar with the procedure can manually generate data for 30 full kinetic curves, composed of ten triplicate points, in 8 h. PMID- 17406498 TI - Immunofluorescence microscopy and immunogold EM for investigating fungal infection of plants. AB - Fungi are a diverse group of eukaryotic organisms whose activities are intricately linked to the lives of human beings. Their involvement in plant productivity, as agents of human diseases, as sources of medicines and enzymes and as model experimental organisms has necessitated the development of sensitive and specific techniques for tracking the organisms and their protein products. Techniques employing highly specific monoclonal antibodies have allowed the visualization of fungi in their natural environments and have facilitated the study of their antigens at the subcellular level. Here, we describe three such techniques, immunofluorescence (IF), immuno-enzymatic staining (IES) and immunoelectron microscopy (IEM), that have found widespread applicability in studies of fungal biology, and which can also be adapted for use in the study of other eukaryotic organisms. Results from the IF and IES procedures can be obtained within 4-5 h. Sample preparation for IEM takes approximately 4 days. Gold labeling and visualization of samples can be completed within 4 h. PMID- 17406499 TI - Collagen antibody-induced arthritis. AB - Collagen antibody-induced arthritis (CAIA) is a simple mouse model of rheumatoid arthritis that can be used to address questions of pathogenic mechanisms and to screen candidate therapeutic agents. Arthritis is stimulated by the administration of a cocktail of monoclonal antibodies that are directed to conserved auto-antigenic epitopes in collagen type II, followed by endotoxin. The antibody-induced arthritis model offers several key advantages over the classic collagen-induced arthritis (CIA) model. These include rapid disease onset, high uptake rate, synchronicity, and the capacity to use genetically modified mice, such as transgenics and knockouts. This protocol takes 1-2 weeks to be completed. PMID- 17406500 TI - Identification and analysis of essential Aspergillus nidulans genes using the heterokaryon rescue technique. AB - In the heterokaryon rescue technique, gene deletions are carried out using the pyrG nutritional marker to replace the coding region of target genes via homologous recombination in Aspergillus nidulans. If an essential gene is deleted, the null allele is maintained in spontaneously generated heterokaryons that consist of two genetically distinct types of nuclei. One nuclear type has the essential gene deleted but has a functional pyrG allele (pyrG+). The other has the wild-type allele of the essential gene but lacks a functional pyrG allele (pyrG-). Thus, a simple growth test applied to the uninucleate asexual spores formed from primary transformants can identify deletions of genes that are non essential from those that are essential and can only be propagated by heterokaryon rescue. The growth tests also enable the phenotype of the null allele to be defined. Diagnostic PCR can be used to confirm deletions at the molecular level. This technique is suitable for large-scale gene-deletion programs and can be completed within 3 weeks. PMID- 17406501 TI - Preparation and maintenance of single-cell micro-island cultures of basal forebrain neurons. AB - Micro-island cultures provide a simplified system for studying the expression of cellular phenotype, excitability, synapse formation and pre- and postsynaptic regulatory mechanisms without the usual problems that arise from complex interactions between large numbers of other cells. The technique relies on the ability to constrain the attachment and growth of either single or small groups of neurons to discrete (20-500 microm) 'islands' of cell-permissive substrate applied over a nonadherent background layer. Constrained in this way, neurons form large numbers of conventional synaptic and/or autaptic contacts that can be easily visualized, making them ideally suited for studying synaptic physiology using electrophysiological and/or high-resolution optical imaging techniques. The protocol described here requires approximately 2 h for preparation of the culture dishes and a further 3-4 h for isolation and plating out the cells. Once established, the cultures can be maintained for prolonged periods (>6 weeks) permitting manipulations to be made to their local environment and the effects on individually identified cells to be repeatedly monitored. PMID- 17406502 TI - Use of a commercially available reagent for the selective detection of homocysteine in plasma. AB - A procedure for the detection of homocysteine (Hcy) in blood plasma is described. A commercially available chromogen is added to the plasma sample. The plasma solution turns from yellow to blue upon heating for 4 min when a detectable threshold level of Hcy is present. Chromatographic separations and immunogenic materials are not needed. The protocol takes approximately 30 min. PMID- 17406503 TI - Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia. AB - This protocol describes the setup, maintenance and characteristics of models of epithelial Candida infections based on well-established three-dimensional organotypic tissues of human oral and vaginal mucosa. Infection experiments are highly reproducible and can be used for the direct analysis of pathogen epithelial cell interactions. This allows detailed investigations of Candida albicans wild type or mutant strain interaction with epithelial tissue or the evaluation of the host immune response using histological, biochemical and molecular methods. As such, the models can be utilized as a tool to investigate cellular interactions or protein and gene expression that are not complicated by non-epithelial factors. To study the impact of innate immunity or the antifungal activity of natural and non-natural compounds, the mucosal infection models can be supplemented with immune cells, antimicrobial agents or probiotic bacteria. The model requires at least 3 days to be established and can be maintained thereafter for 2-4 days. PMID- 17406504 TI - A protocol for preparing, characterizing and using three RNA-specific, live cell imaging probes: E36, E144 and F22. AB - This protocol outlines a methodology for the preparation and characterization of three RNA-specific fluorescent probes (E36, E144 and F22) and their use in live cell imaging. It describes a detailed procedure for their chemical synthesis and purification; serial product characterization and quality control tests, including measurements of their fluorescence properties in solution, measurement of RNA specificity and analysis of cellular toxicity; and live cell staining and counterstaining with Hoechst or DAPI. Preparation and application of these RNA imaging probes takes 1 week. PMID- 17406505 TI - Isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. AB - Isolation of transcriptionally active nuclei from plant tissues is a fundamental first step in many plant molecular biology protocols. Enriched nuclear fractions may be used in "run-on" assays to measure the rate of transcription for any given gene, adding additional resolution to assays of steady-state transcript accumulation such as RNA-gel blots, RT-PCR or microarrays. The protocols presented here streamline, adapt and optimize existing methods for use in Arabidopsis thaliana. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. This capability complements the immense body of steady-state transcript measurements and indirectly identifies instances where message turnover may have a critical and/or primary role in regulating gene expression levels. PMID- 17406506 TI - Using circular dichroism collected as a function of temperature to determine the thermodynamics of protein unfolding and binding interactions. AB - Circular dichroism (CD) is an excellent spectroscopic technique for following the unfolding and folding of proteins as a function of temperature. One of its principal applications is to determine the effects of mutations and ligands on protein and polypeptide stability. If the change in CD as a function of temperature is reversible, analysis of the data may be used to determined the van't Hoff enthalpy and entropy of unfolding, the midpoint of the unfolding transition and the free energy of unfolding. Binding constants of protein-protein and protein-ligand interactions may also be estimated from the unfolding curves. Analysis of CD spectra obtained as a function of temperature is also useful to determine whether a protein has unfolding intermediates. Measurement of the spectra of five folded proteins and their unfolding curves at a single wavelength requires approximately 8 h. PMID- 17406507 TI - A simple trogocytosis-based method to detect, quantify, characterize and purify antigen-specific live lymphocytes by flow cytometry, via their capture of membrane fragments from antigen-presenting cells. AB - We have developed a method exploiting the phenomenon of trogocytosis to detect lymphocytes reacting specifically with target cells by flow cytometry. Trogocytosis is a process by which lymphocytes capture fragments of the plasma membrane from the antigen-presenting cells (APCs) expressing their cognate antigen. For this method, a label (such as a fluorescent lipid or biotin) is first incorporated in the membrane of APCs. These labeled cells are then co cultured for a few hours with a population of cells containing the lymphocytes to be detected. After this period of stimulation, lymphocytes that have performed trogocytosis are identified by their acquisition of the label initially present on the APC membrane using flow cytometry. A major advantage of this method is its compatibility with the simultaneous detection of phenotypic and/or functional markers on the lymphocytes. Furthermore, cells can be recovered alive and active after detection of trogocytosis, and are therefore available for further characterization or even conceivably for therapeutic purposes. PMID- 17406508 TI - The mode-of-action by network identification (MNI) algorithm: a network biology approach for molecular target identification. AB - This protocol details the use of the mode-of-action by network identification (MNI) algorithm to identify the gene targets of a drug treatment based on gene expression data. Investigators might also use the MNI algorithm to identify the gene mediators of a disease or the physiological state of cells and tissues. The MNI algorithm uses a training data set of hundreds of expression profiles to construct a statistical model of gene-regulatory networks in a cell or tissue. The model describes combinatorial influences of genes on one another. The algorithm then uses the model to filter the expression profile of a particular experimental treatment and thereby distinguish the molecular targets or mediators of the treatment response from hundreds of additional genes that also exhibit expression changes. It takes approximately 1 h per run, although run time is significantly affected by the size of the genome and data set. PMID- 17406509 TI - 5' end cDNA amplification using classic RACE. AB - The 5' ends of transcripts provide important information about transcription initiation sites and the approximate locations of local cis-acting enhancer elements; it is therefore important to establish the 5' ends with some precision. RACE (rapid amplification of cDNA ends) PCR is useful for quickly obtaining full length cDNAs for mRNAs for which only part of the sequence is known and to identify alternative 5' or 3' ends of fully sequenced genes. The method consists of using PCR to amplify, from complex mixtures of cellular mRNA, the regions between the known parts of the sequence and non-specific tags appended to the ends of the cDNA. Whereas the poly(A) tail serves to provide such a tag at the 3' end of the mRNA, an artificial one needs to be generated at the 5' end, and various approaches have been described to address this step. The classical scheme for 5' RACE described here is simple, suffices in many instances in which RACE is needed and can be performed in 1-3 days. PMID- 17406510 TI - Analysis of mitochondrial function using phosphorescent oxygen-sensitive probes. AB - Mitochondrial dysfunction has been associated with a variety of currently marketed therapeutics and has also been implicated in many disease states. Alterations in the rate of oxygen consumption are an informative indicator of mitochondrial dysfunction, but the use of such assays has been limited by the constraints of traditional measurement approaches. Here, we present a high throughput, fluorescence-based methodology for the analysis of mitochondrial oxygen consumption using a phosphorescent oxygen-sensitive probe, standard microtitre plates and plate reader detection. The protocol describes the isolation of mitochondria from animal tissue, initial establishment and optimization of the oxygen consumption assay, subsequent screening of compounds for mitochondrial toxicity (uncoupling and inhibition), data analysis and generation of dose-response curves. It allows dozens of compounds (or hundreds of assay points) to be analyzed in a single day, and can be further up-scaled, automated and adapted for other enzyme- and cell-based screening applications. PMID- 17406511 TI - Single nucleotide polymorphism (SNP) allele frequency estimation in DNA pools using Pyrosequencing. AB - Identifying the genetic variation underlying complex disease requires analysis of many single nucleotide polymorphisms (SNPs) in a large number of samples. Several high-throughput SNP genotyping techniques are available; however, their cost promotes the use of association screening with pooled DNA. This protocol describes the estimation of SNP allele frequencies in pools of DNA using the quantitative sequencing method Pyrosequencing (PSQ). PSQ is a relatively recently described high-throughput method for genotyping, allele frequency estimation and DNA methylation analysis based on the detection of real-time pyrophosphate release during synthesis of the complementary strand to a PCR product. The protocol involves the following steps: (i) quantity and quality assessment of individual DNA samples; (ii) DNA pooling, which may be undertaken at the pre- or post-PCR stage; (iii) PCR amplification of PSQ template containing the variable sequence region of interest; and (iv) PSQ to determine the frequency of alleles at a particular SNP site. Once the quantity and quality of individual DNA samples has been assessed, the protocol usually requires a few days for setting up pre PCR pools, depending on sample number. After PCR amplification, preparation and analysis of PCR amplicon by PSQ takes 1 h per plate. PMID- 17406512 TI - A protocol for mosaic analysis with a repressible cell marker (MARCM) in Drosophila. AB - Mosaic analysis with a repressible cell marker (MARCM) is a genetic technique used in Drosophila to label single cells or multiple cells sharing a single progenitor. Labeled homozygous mutant cells can be generated in an otherwise unlabeled heterozygous animal. Mutant or wild-type labeled cells can also be made to express one or more transgenes. Major applications of MARCM include (i) lineage analysis, (ii) investigating gene function in single or small populations of cells and (iii) neuronal circuit tracing. Our laboratory uses MARCM primarily to label and genetically manipulate neurons; however, this protocol can be adapted to any cell of interest. The protocol involves generating two fly stocks with the necessary genetic elements for MARCM analysis and subsequently generating MARCM clones. Labeled clones can be followed in live and fixed tissues for high-resolution analysis of wild-type or genetically manipulated cells.NOTE: In the PDF version of this article initially published online, the first "FRT" and the "Mutation" labels in Figure 1b were transposed. In both the PDF and HTML versions, "mutant" was omitted from the label on the right, which should read "Labeled homozygous mutant daughter cell". The figure has been corrected in all versions of the article. PMID- 17406513 TI - Preparation of substituted anilines from nitro compounds by using supported gold catalysts. AB - A protocol for the chemoselective hydrogenation of nitro compounds to the corresponding anilines by means of supported gold catalysts is described. Nitro groups on different compounds--containing double bonds, carbonyl, nitrile or amide groups--have been successfully hydrogenated on supported gold nanoparticles (Au/TiO2 and Au/Fe2O3), using a batch reactor under H2 pressure. Unlike other noble metals, gold shows high chemoselectivity towards reduction of the nitro group, at near-complete conversion of the substrate. The total time to carry out this protocol strongly depends on the reaction step, which is a function of the activity of the catalyst and the nature of the substrate. PMID- 17406514 TI - Total synthesis of multi-kilobase DNA sequences from oligonucleotides. AB - A method for synthesizing DNA from 40-mer oligonucleotides, which we used to generate a 32-kb DNA fragment, is explained. DNA sequences are synthesized as approximately 500 bp fragments (synthons) in a two-step PCR reaction and cloned using ligation-independent cloning (LIC). Synthons are then assembled into longer full-length sequences in a stepwise manner. By initially synthesizing smaller fragments (synthons), the number of clones sequenced is low compared with synthesizing complete multi-kilobase DNA sequences in a single step. LIC eliminates the need for purification of fragments before cloning, making the process amenable to high-throughput operation and automation. Type IIs restriction enzymes allow seamless assembly of synthons without placing restrictions on the sequence being synthesized. Synthetic fragments are assembled in pairs to generate the final construct using vectors that allow selection of desired clones with two unique antibiotic resistance markers, and this eliminates the need for purification of fragments after digestion with restriction endonucleases. PMID- 17406515 TI - Single-tube reaction using peptide nucleic acid as both PCR clamp and sensor probe for the detection of rare mutations. AB - The detection of rare mutant DNA from a background of wild-type alleles usually requires laborious manipulations, such as restriction enzyme digestion and gel electrophoresis. Here, we describe a protocol for homogeneous detection of rare mutant DNA in a single tube. The protocol uses a peptide nucleic acid (PNA) as both PCR clamp and sensor probe. The PNA probe binds tightly to perfectly matched wild-type DNA template but not to mismatched mutant DNA sequences, which specifically inhibits the PCR amplification of wild-type alleles without interfering with the amplification of mutant DNA. A fluorescein tag (which undergoes fluorescence resonance energy transfer with the adjacent fluorophore of an anchor probe when both are annealed to the template DNA) also allows the PNA probe to generate unambiguous melting curves to detect mutant DNA during real time fluorescent monitoring. The whole assay takes about only 1 h. This protocol has been used for detecting mutant K-ras DNA and could be applied to the detection of other rare mutant DNAs. PMID- 17406516 TI - Highly purified selective isolation of eosinophils from human peripheral blood by negative immunomagnetic selection. AB - Eosinophils are a minority constituent in human peripheral blood. The study of eosinophils has been limited by difficulty in achieving sufficient cell number and purity. We describe a modified protocol for immunomagnetic cell separation for efficient isolation of human peripheral blood eosinophils. We employ a mixture of monoclonal antibodies (mAbs) directed against cell-surface antigens on human hematopoietic cells combined with secondary labeling with a colloidal suspension of magnetic dextran-iron particles for negative selection of eosinophils. Unwanted labeled cells are retained in the magnetized column, permitting high recovery (70%) and purity (>98%) of eosinophils while retaining cell viability. Eosinophils remain quiescent after isolation, and stimulation caused by cytokines upregulates (i) cell-cell or cell-extracellular matrix protein adhesion, (ii) secretion of bioactive mediators and (iii) cell-surface adhesion molecules. This method for purified isolation is accomplished in < or = 4 h and preserves eosinophils in a quiescent, viable state. PMID- 17406517 TI - Cell cycle synchronization of tobacco BY-2 cells. AB - Synchronization is a powerful technique for understanding cell cycle events. Here, we describe the procedure for synchronizing tobacco bright yellow 2 (BY-2) cell line, with which an exceptionally high level of synchrony can be achieved. It basically relies on an "arrest-and-release" strategy using aphidicolin, an inhibitor of DNA replication, and propyzamide, a plant-microtubule disruptant. In a single-step process using aphidicolin alone, a cell population with about 70% of the cells at mitosis can be achieved, whereas by a two-step method using the two inhibitors sequentially, the level of synchrony can reach over 90%. The method of choice depends not only on the peak mitotic cell proportion but also on the cell cycle stage that is targeted for analysis. Both procedures take about 1.5 days, and cell cycle progression can be observed from the S phase to the next G1 phase at about 12 h after a 24 h-period treatment with aphidicolin. PMID- 17406518 TI - Imaging the lateral diffusion of membrane molecules with quantum dots. AB - This protocol describes a sensitive approach to tracking the motion of membrane molecules such as lipids and proteins with molecular resolution in live cells. This technique makes use of fluorescent semiconductor nanocrystals, quantum dots (QDs), as a probe to detect membrane molecules of interest. The photostability and brightness of QDs allow them to be tracked at a single particle level for longer periods than previous fluorophores, such as fluorescent proteins and organic dyes. QDs are bound to the extracellular part of the object to be followed, and their movements can be recorded with a fluorescence microscope equipped with a spectral lamp and a sensitive cooled charge-coupled device camera. The experimental procedure described for neurons below takes about 45 min. This technique is applicable to various cultured cells. PMID- 17406519 TI - Immunofluorescent staining and imaging of the pupal and adult Drosophila visual system. AB - This immunofluorescence protocol can be used to assay cell morphology, cell positioning and subcellular localization of proteins in the fly eye at stages of development from early pupation to adult. The protocol includes the following procedures: collecting and developmentally staging Drosophila pupae, dissecting fly eyes at defined stages of development, immunostaining of retina and preparing visual system samples (i.e., retina and optic lobe) for confocal microscopy. It is supplemented with images of key dissection steps, guidelines for troubleshooting and examples of data obtained using these methods. Overall, this protocol takes up to 9 d to complete. The amount of hands-on time required on each day varies, ranging from 30 min to several hours depending on the number of stages and/or genotypes one wishes to study. PMID- 17406520 TI - Primary culture of ovarian surface epithelial cells and ascites-derived ovarian cancer cells from patients. AB - Our laboratory has refined the technique for isolating primary cultures of normal human ovarian surface epithelial (OSE) cells by combining two different protocols involving the enzymatic and mechanical removal of OSE cells from ovarian biopsies. A simple protocol of obtaining primary epithelial ovarian cancer (EOC) cells from the ascites fluid removed from patients with high-grade ovarian cancer is also described. These methods allow for the direct application of many molecular and cellular analyses of normal versus cancer cells isolated freshly from patients, with the added potential for retrospective analyses of archived cells and tissues. Thus, we have included optional steps for the immediate preparation of ascites-derived EOC cells to be used for subsequent cytological analyses. Initial isolation of OSE or EOC cells can be completed in 1 h, and primary cells are further expanded in culture for several weeks. PMID- 17406521 TI - A practical recipe for stable isotope labeling by amino acids in cell culture (SILAC). AB - Stable isotope labeling by amino acids in cell culture (SILAC) is a simple, robust, yet powerful approach in mass spectrometry (MS)-based quantitative proteomics. SILAC labels cellular proteomes through normal metabolic processes, incorporating non-radioactive, stable isotope-containing amino acids in newly synthesized proteins. Growth medium is prepared where natural ("light") amino acids are replaced by "heavy" SILAC amino acids. Cells grown in this medium incorporate the heavy amino acids after five cell doublings and SILAC amino acids have no effect on cell morphology or growth rates. When light and heavy cell populations are mixed, they remain distinguishable by MS, and protein abundances are determined from the relative MS signal intensities. SILAC provides accurate relative quantification without any chemical derivatization or manipulation and enables development of elegant functional assays in proteomics. In this protocol, we describe how to apply SILAC and the use of nano-scale liquid chromatography coupled to electrospray ionization mass spectrometry for protein identification and quantification. This procedure can be completed in 8 days. PMID- 17406522 TI - Advances in water treatment by adsorption technology. AB - Among various water purification and recycling technologies, adsorption is a fast, inexpensive and universal method. The development of low-cost adsorbents has led to the rapid growth of research interests in this field. The present protocol describes salient features of adsorption and details experimental methodologies for the development and characterization of low-cost adsorbents, water treatment and recycling using adsorption technology including batch processes and column operations. The protocol describes the development of inexpensive adsorbents from waste materials, which takes only 1-2 days, and an adsorption process taking 15-120 min for the removal of pollutants. The applications of batch and column processes are discussed, along with suggestions to make this technology more popular and applicable. PMID- 17406523 TI - Multiplex PCR-based reverse line blot hybridization assay (mPCR/RLB)--a practical epidemiological and diagnostic tool. AB - Combining multiplex PCR, sequentially, with reverse line blot hybridization (mPCR/RLB) is a convenient, objective way to identify up to 43 targets in 43 individual specimens simultaneously (using a 45-lane membrane format). It is more flexible and less expensive than DNA microarray. The number of targets is adequate for epidemiological and most clinical diagnostic applications; based on the same target (43) and specimen numbers (43), it is much more practical than conventional uniplex PCR (uPCR) and mPCR. We have used the protocol to identify and subtype bacteria, viruses and fungi and identify pathogens in clinical specimens; potentially, it could be used for many other applications, such as detection of mutations in, or identification of alleles of, eukaryotic genes. Development of each assay involves (i) careful primer and probe design, based on literature and sequence database searches, which are critical to success of the assay; and (ii) bench-top evaluation, using known samples, controls and dilution series, to confirm sensitivity, specificity and reproducibility. The assay takes about one and half working days to complete; about 4 h for the mPCR and 6 h for the RLB, including a total of 4 h 'hands-on' time. PMID- 17406524 TI - Preparation of primary cultured mesenteric artery smooth muscle cells for fluorescent imaging and physiological studies. AB - In this protocol, we describe a method for isolation and culture of smooth muscle cells derived from the adult rat (or mouse) superior mesenteric artery. Arterial myocytes are obtained by enzymatic dissociation and established in primary culture. The cultured cells retain expression of smooth muscle-specific alpha actin and physiological responses to agonists. Cultured arterial myocytes (prepared from wild-type or transgenic animals) provide a useful model for studying the regulation of a wide range of vascular smooth muscle responses at the cellular and subcellular levels. Plasmids, RNA interference and antisense oligodeoxynucleotides can be readily introduced into the cells to alter protein expression. Fluorescent dyes can also be introduced to visualize a variety of activities, some of which may be specific to vascular smooth muscle cells. This protocol requires about 3 h on each of 2 consecutive days to complete. PMID- 17406525 TI - Use of a lentivirus/VSV pseudotype virus for highly efficient genetic redirection of human peripheral blood lymphocytes. AB - Genetic redirection of lymphocytes that have been engineered to recognize antigens other than those originally programmed by their rearranged germlines is a potentially powerful immunotherapeutic tool. The rationale for the protocol described here is that many cancers and persistent or latent viruses have developed strikingly similar mechanisms of evading attack by host immunity that can often be overcome by redirection of host lymphocytes using chimeric T-cell receptor (chTCR) genes. However, for human peripheral blood lymphocytes (PBLs), this is generally regarded as a technically demanding procedure with unacceptably low efficiency using either contemporary transfection methods or retroviral transduction. One of the main difficulties with retroviruses is their reliance on rapidly dividing cells for integration of their genomes carrying the desired chTCR. Here we describe a highly efficient protocol that uses a lentivirus/vesicular stomatitis virus pseudotyped virus to engineer CD3/CD28 stimulated human peripheral blood cells (i.e., primarily T cells), with near 100% efficiency. PMID- 17406526 TI - Investigating macromolecules inside cultured and injected cells by in-cell NMR spectroscopy. AB - The noninvasive character of NMR spectroscopy, combined with the sensitivity of the chemical shift, makes it ideally suited to investigate the conformation, binding events and dynamics of macromolecules inside living cells. These 'in-cell NMR' experiments involve labeling the macromolecule of interest with a nonradioactive but NMR-active isotope (15N or 13C). Cellular samples are prepared either by selectively overexpressing the protein in suitable cells (e.g., bacterial cells grown on isotopically labeled media), or by injecting isotopically labeled proteins directly into either cells or cell extracts. Here we provide detailed protocols for in-cell NMR experiments in the prokaryotic organism Escherichia coli, as well as eukaryotic cells and extracts employing Xenopus laevis oocytes or egg extracts. In-cell NMR samples with proteins overexpressed in E. coli can be produced within 13-14 h. Preparing Xenopus oocyte samples for in-cell NMR experiments takes 6-14 h depending on the oocyte preparation scheme and the injection method used. PMID- 17406527 TI - Preparation and square wave electroporation of retinal explant cultures. AB - This protocol details organotypic cultures of developing mouse, monkey and human retinas, which can be maintained for up to 2 weeks. Intact retinas are placed on polycarbonate filters floating on explant culture medium and fed every day with previously prepared retinal conditioned medium. Developing mouse retinas from E12.5 to P12 have been successfully cultured using this protocol as well as retinas from the equivalent stages of human and monkey development. Although this protocol does not require any special equipment, it provides a relatively high throughput. Retinal explant cultures lend themselves to complex pharmacological and genetic manipulations that are currently not feasible in vivo. A detailed procedure for square wave electroporation of retinal explants is also included to provide a high-throughput means to alter gene expression in the developing retina. This protocol for the preparation of retinal conditioned explant medium requires 4 d. Other steps of this protocol can be completed in 2 h. PMID- 17406528 TI - Immunohistochemical staining of post-mortem adult human brain sections. AB - One of the challenges for modern neuroscience is to understand the basis of coordinated neuronal function and networking in the human brain. Some of these questions can be addressed using low- and high-resolution imaging techniques on post-mortem human brain tissue. We have established a versatile protocol for fixation of post-mortem adult human brain tissue, storage of the tissue in a human brain bank, and immunohistochemical analysis in order to understand human brain functions in normal controls and in neuropathological conditions. The brains are fixed by perfusion through the internal carotid and basilar arteries to enhance the penetration of fixative throughout the brain, then blocked, postfixed, cryoprotected, snap-frozen and stored at -80 degrees C. Sections are processed for immunohistochemical single- or double-label staining and conventional-, electron- or confocal laser scanning-microscopy analysis. The results gained using this tissue and protocol are vital for determining the localization of neurochemicals throughout the human brain and to document the changes that occur in neurological diseases. PMID- 17406529 TI - Determination of the folding of proteins as a function of denaturants, osmolytes or ligands using circular dichroism. AB - Circular dichroism (CD) is an excellent tool for examining the interactions and stability of proteins. This protocol covers methods to obtain and analyze circular dichroism spectra to measure changes in the folding of proteins as a function of denaturants, osmolytes or ligands. Applications include determination of the free energy of folding of a protein, the effects of mutations on protein stability and the estimation of binding constants for the interactions of proteins with other proteins, DNA or ligands, such as substrates or inhibitors. The experiments require 2-5 h. PMID- 17406530 TI - 3' end cDNA amplification using classic RACE. AB - Having knowledge of the entire 3' sequence of a cDNA is often important because the non-coding terminal region can contain signals that regulate the stability or subcellular localization of the mRNA. Also, some messages use alternative genomic sites for cleavage and polyadenylation that can alter the above properties, or change the encoded protein. Full-length cDNAs can be obtained from complex mixtures of cellular mRNA using rapid amplification of cDNA ends (RACE) PCR as long as part of the mRNA sequence is known; adding non-specific tags to the ends of the cDNA allows the regions between the known parts of the sequence and the ends to be amplified. In 3' RACE, the poly(A) tail functions as a non-specific tag at the 3' end of the mRNA. cDNA ends can be obtained in 1-3 days using this protocol. PMID- 17406531 TI - Sequence-specific amplification polymorphisms (SSAPs): a multi-locus approach for analyzing transposon insertions. AB - Multiple copies of transposable elements, inserted at random around the host genome, can be used as molecular markers. Sequence-specific amplification polymorphisms (SSAPs) amplify the region between a PCR primer site near the end of an element and an adjacent restriction site in the flanking genomic DNA. Each amplified insertion is revealed as a band on a sequencing gel or capillary electrophoresis, and a genomic DNA sample produces a characteristic fingerprint of bands. Here, we explain the SSAP marker development method, which can be performed for any species, with recommendations for experimental parameters in several plant species. It takes about 1-2 weeks to complete the whole SSAP marker procedure. PMID- 17406532 TI - Preparation of ready-to-use, storable and reconstituted type I collagen from rat tail tendon for tissue engineering applications. AB - Collagen is a widely investigated extracellular matrix material with extensive potentials in the field of tissue engineering. This protocol describes a method to prepare reconstituted collagen that can be ready-to-use, storable and suitable for further in vitro and in vivo investigations. Type I collagen was extracted from rat tail tendons and processed in acetic acid solution to obtain sterile soluble collagen. At first, crude collagen was dissolved in acetic acid, then frozen at -20 degrees C and lyophilized to obtain a sponge, which could be stored at -80 degrees C. Lyophilized collagen was then dispersed in acetic acid to obtain a sterile solution of collagen at targeted concentrations. The whole low cost process from the extraction to the final sterile solution takes around 2-3 weeks. The collagen solution, once neutralized, has the potential to be used to produce gels or scaffolds, to deposit thin films on supports and to develop drug delivery systems. PMID- 17406533 TI - Long-term primary culture of secretory cells of Bothrops jararaca venom gland for venom production in vitro. AB - This protocol details the optimal conditions to establish a long-term primary culture of secretory cells from the venom gland of the Bothrops jararaca snake. Furthermore, these conditions allow the production and secretion of venom into the culture medium. Snake venom is a rich source of active molecules and has been used for bioprospection studies. However, obtaining enough venom from snakes is a major obstacle. Secretory cells of venom glands are capable of producing active toxins. Therefore, a culture of secretory cells is a good in vitro system to acquire the venom of snakes without capturing the animal from the wild. The protocol described here provides a rapid (approximately 4 h) and reproducible means of producing sufficient amounts of snake venom for biological investigations. PMID- 17406534 TI - Restriction landmark genomic scanning. AB - Restriction landmark genomic scanning (RLGS) is a method to detect large numbers of restriction landmarks in a single experiment. It is based on the concept that restriction enzyme sites can serve as landmarks throughout a genome. RLGS uses direct end-labeling of the genomic DNA digested with a rare-cutting restriction enzyme and high-resolution two-dimensional electrophoresis. Compared with the conventional gene-detection technologies, such as Southern blot analysis and PCR, RLGS has the following advantages even though it needs specially designed instruments: high-efficiency scanning capacity, scanning extensibility by using alternate restriction enzyme combinations, applicability to any organism, a spot intensity that reflects the copy number of restriction landmarks, and the ability, by using a methylation-sensitive enzyme, to screen the methylated state of genomic DNA. The RLGS protocol can be accomplished in 5 days to 2 weeks. PMID- 17406535 TI - Identification of membrane proteins from mammalian cell/tissue using methanol facilitated solubilization and tryptic digestion coupled with 2D-LC-MS/MS. AB - The core prerequisites for an efficient proteome-scale analysis of mammalian membrane proteins are effective isolation, solubilization, digestion and multidimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS). This protocol is for analysis of the mammalian membrane proteome that relies on solubilization and tryptic digestion of membrane proteins in a buffer containing 60% (vol/vol) methanol. Tryptic digestion is followed by strong cation exchange (SCX) chromatography and reversed phase (RP) chromatography coupled online with MS/MS for protein identification. The use of a methanol-based buffer eliminates the need for reagents that interfere with chromatographic resolution and ionization of the peptides (e.g., detergents, chaotropes, inorganic salts). Sample losses are minimized because solubilization and digestion are carried out in a single tube avoiding any sample transfer or buffer exchange between these steps. This protocol is compatible with stable isotope labeling at the protein and peptide level, enabling identification and quantitation of integral membrane proteins. The entire procedure--beginning with isolated membrane fraction and finishing with MS data acquisition--takes 4-5 d. PMID- 17406536 TI - Biochemical assays for multiple activation states of protein kinase C. AB - This protocol describes biochemical procedures to monitor the activation of the protein kinase C (PKC) family using PKCdelta as the representative. The PKC family is composed of ten isoforms divided into cPKC, nPKC and aPKC groups, and their catalytic activity is regulated by multiple mechanisms. For example, PKCdelta in the nPKC group is activated by diacylglycerol as a second messenger in the receptor-coupled manner, through tyrosine phosphorylation and protein complex formation in stress-stimulated cells, and by the caspase-catalyzed cleavage during apoptosis. The isoform is immunoprecipitated from cultured cells, the protein kinase activity is measured by in vitro kinase assay and the tyrosine phosphorylation and protein complex formation are characterized by immunoblot, whereas the generation of the catalytic fragment is detected by immunoblot in the cell extract. The combination of these procedures is useful to evaluate the activation states of the PKC family in cells. This protocol can be completed in 3 5 d. PMID- 17406537 TI - A protocol for Agrobacterium-mediated transformation in rice. AB - Agrobacterium-mediated transformation of rice is an important method that has been widely adopted by many laboratories. However, because current approaches rely on culture systems, routine protocols have been established only in japonica rice, especially those varieties with higher regeneration potential. Some very efficient methods have been developed for japonica varieties that enable high throughput functional analysis in rice; however, many elite japonica, and most indica, varieties are difficult to regenerate, leading to low transformation efficiencies. Much effort has been devoted to improving transformation efficiency for all rice genotypes. Here, we describe an Agrobacterium-mediated rice transformation method that is applicable to easily cultured varieties in addition to elite japonica varieties that are more difficult to culture. Using this method, transgenic rice plants can be obtained in about 2-3 months with a transformation frequency of 30-50%, both in easily cultured varieties and recalcitrant elite japonica rice. PMID- 17406538 TI - Isolation of skin-derived precursors (SKPs) and differentiation and enrichment of their Schwann cell progeny. AB - This protocol describes methods of isolating skin-derived precursors (SKPs) from rodent and human skin, and for generating and enriching Schwann cells from rodent SKPs. SKPs are isolated as a population of non-adherent cells from the dermis that proliferate and self-renew as floating spheres in response to fibroblast growth factor 2 (FGF2) and epidermal growth factor (EGF). Their differentiation into Schwann cells and subsequent enrichment of these differentiated progeny involves culturing SKPs as adherent cells in the absence of FGF2 and EGF, but in the presence of neuregulins, and then mechanically isolating the Schwann cell colonies using cloning cylinders. Methods for expanding and characterizing these Schwann cells are provided. Generation of primary SKPs takes approximately 2 weeks, while differentiation of Schwann cells requires an additional 4-6 weeks. PMID- 17406539 TI - Electron cryotomography sample preparation using the Vitrobot. AB - Electron cryotomography is the highest-resolution structural technique currently available that can be applied to unique objects such as flexible large protein complexes, irregular viruses, organelles and small cells. Specimens are preserved in a near-native, 'frozen-hydrated' state by vitrification. The thickness of the vitreous ice must be optimized for each specimen, and gold fiducials are typically added to facilitate image alignment. Here, we describe in detail our protocols for electron cryotomography sample preparation including (i) introduction of fiducial markers into the sample and (ii) sample vitrification. Because we almost exclusively use an automated, climate-controlled plunge freezing device (the FEI Vitrobot) to vitrify our samples, we discuss its operation and parameters in detail. A session in which eight grids are prepared takes 1.5-2 h. PMID- 17406540 TI - An optimized protocol for protein purification in cultured mammalian cells using a tandem affinity purification approach. AB - This protocol describes a method that we developed to adapt the tandem affinity purification (TAP) approach for use in mammalian cells. The protocol involves fusing a protein of interest with a tandem tag consisting of two FLAG tags (FF) followed by two protein-A immunoglobulin G (IgG) binding domains (ZZ). The protocol improves upon previously published TAP approaches by employing FLAG in place of calmodulin binding peptide (CBP) with resulting higher recovery during purification. In addition, we use a bicistronic expression system that ensures recovery of stably transfected cell lines expressing easily detectable levels of the protein of interest. A method is also presented for generating cytoplasmic and nuclear extracts, which extends use of this protocol to identify protein protein interactions occurring specifically in the cytoplasm or nucleus. This protocol facilitates the preparation of partially purified recombinant protein and identification of protein-protein interactions in mammalian cell culture models. The protocol can be completed in 34 h. PMID- 17406541 TI - Assessing hoarding in mice. AB - Hoarding is a species-typical behavior shown by rodents, as well as other animals. By hoarding, the rodent secures a food supply for times of emergency (for example, when threatened by a predator) or for times of seasonal adversity such as winter. Scatter hoarding, as seen typically in squirrels and birds, involves placing small caches of food in hidden places, generally underground. Most rodents, however, hoard a supply of food in or near the home base--for example, in 'larders' near the sleeping quarters in a burrow. In the laboratory, measurement of hoarding involves simply weighing the food transported into the home cage from an external source, but the route to that source must be secure and animal-proof; for example, there should be no holes large enough to permit escape of a mouse, and no weak points that could be enlarged by gnawing. A suitable and easily constructed apparatus is described in the protocol. Hoarding has been shown to be sensitive to brain lesions and pharmacological agents, and is a suitable test for species-typical behavior in genetically modified mice. PMID- 17406542 TI - Quantitative multiplex degenerate PCR for human endogenous retrovirus expression profiling. AB - Expression of human endogenous retroviruses (HERV) has been recurrently observed during cellular differentiation or transformation processes in both cell culture and in vivo. Quantitative approaches that analyze variations in HERV transcription could therefore be valuable for cancer diagnosis. We have developed a quantitative assay combining multiplex degenerate PCR (MD-PCR) and a colorimetric Oligo Sorbent Array (OLISA). Quantification of the expression of these multifamily genes relies on the optimization of the amplification primer mix, that is, the primer degeneracy, the relative concentration of each primer and the total amount of primer. Amplification products of each of the nine studied HERV families are independently and specifically detected and quantified using the OLISA microarray. This method constitutes an improvement over previous pan-retrovirus amplification-based methods, which are mainly qualitative. Furthermore, as MD-PCR/OLISA simultaneously monitors several HERV families, it challenges single-family quantitative RT-PCR. Last, the protocol below provides general rules for the design of MD-PCR applications. Once primers have been designed and optimized, the procedure can be completed in 2 days. PMID- 17406543 TI - ChIP-on-chip protocol for genome-wide analysis of transcription factor binding in Drosophila melanogaster embryos. AB - This protocol describes a method to detect in vivo associations between proteins and DNA in developing Drosophila embryos. It combines formaldehyde crosslinking and immunoprecipitation of protein-bound sequences with genome-wide analysis using microarrays. After crosslinking, nuclei are enriched using differential centrifugation and the chromatin is sheared by sonication. Antibodies specifically recognizing wild-type protein or, alternatively, a genetically encoded epitope tag are used to enrich for specifically bound DNA sequences. After purification and polymerase chain reaction-based amplification, the samples are fluorescently labeled and hybridized to genomic tiling microarrays. This protocol has been successfully used to study different tissue-specific transcription factors, and is generally applicable to in vivo analysis of any DNA binding proteins in Drosophila embryos. The full protocol, including the collection of embryos and the collection of raw microarray data, can be completed within 10 days. PMID- 17406544 TI - In-gel digestion for mass spectrometric characterization of proteins and proteomes. AB - In-gel digestion of proteins isolated by gel electrophoresis is a cornerstone of mass spectrometry (MS)-driven proteomics. The 10-year-old recipe by Shevchenko et al. has been optimized to increase the speed and sensitivity of analysis. The protocol is for the in-gel digestion of both silver and Coomassie-stained protein spots or bands and can be followed by MALDI-MS or LC-MS/MS analysis to identify proteins at sensitivities better than a few femtomoles of protein starting material. PMID- 17406545 TI - Synthesis of a biocleavable polyrotaxane-plasmid DNA (pDNA) polyplex and its use for the rapid nonviral delivery of pDNA to cell nuclei. AB - This protocol provides a method for synthesizing a biocleavable polyrotaxane/plasmid DNA (pDNA) polyplex and for using it to deliver pDNA into cell nuclei. The biocleavable polyrotaxane is synthesized in four steps: (i) introduction of disulfide linkages at both terminals of PEG, (ii) preparation of an inclusion complex between disulfide-containing PEG and alpha-cyclodextrins (alpha-CDs), (iii) synthesis of polyrotaxane and (iv) modification of alpha-CDs in the polyrotaxane with dimethylethylenediamine. A polyplex of pDNA with the polyrotaxane is formed when the two compounds are dissolved together in a phosphate buffer. Subcellular localization of rhodamine-labeled pDNA in fluorescently labeled organelles is quantified by Z-series of confocal images captured by confocal laser scanning microscopy. Significant amounts of pDNA delivered to the nucleus can be expected as well as high transfection activity of the polyplex. This protocol can be completed in 23-32 d. PMID- 17406546 TI - Isolation of CD4+ T cells from murine lungs: a method to analyze ongoing immune responses in the lung. AB - The regulation of the cellular immune response in lung diseases is not yet fully understood. Isolating different subsets of immune cells directly from the lung is therefore an indispensable method of gaining detailed knowledge on the function of these cells in this organ. This protocol describes a method of isolating and magnetically labeling CD4+ lung T cells, which are then loaded and retained on the column while all other cells run through it (positive selection). The yield of this isolation is approximately 5 x 10(5) to 1.5 x 10(6) CD4+ cells from a murine lung. These cells can be further investigated by several methods such as flow cytometry, western blot analysis, RT-PCR, immunostaining and ELISA. In addition, lung CD4+ T cells alone or along with other immunologically important cells such as CD8+ T cells and T regulatory cells can be adoptively transferred into immune-deficient mice, and can influence important local parameters. This protocol can be completed in approximately 4 h 20 min. PMID- 17406547 TI - Using circular dichroism spectra to estimate protein secondary structure. AB - Circular dichroism (CD) is an excellent tool for rapid determination of the secondary structure and folding properties of proteins that have been obtained using recombinant techniques or purified from tissues. The most widely used applications of protein CD are to determine whether an expressed, purified protein is folded, or if a mutation affects its conformation or stability. In addition, it can be used to study protein interactions. This protocol details the basic steps of obtaining and interpreting CD data, and methods for analyzing spectra to estimate the secondary structural composition of proteins. CD has the advantage that measurements may be made on multiple samples containing < or =20 microg of proteins in physiological buffers in a few hours. However, it does not give the residue-specific information that can be obtained by x-ray crystallography or NMR. PMID- 17406549 TI - High resolution colonoscopy in live mice. AB - Endoscopy in humans is a powerful method for physicians to examine the gut for inflammatory or neoplastic changes. In medical and immunological research, animal models of intestinal diseases are established key tools to investigate the mucosal immune system, colitis and cancer development in the gut. Moreover, such models represent valid systems for testing of novel drugs. In the past, mice had to be killed in order to analyze colitis activity and tumor development. The following protocol describes a method to perform high resolution endoscopic monitoring of live mice. Mice developing colitis or colonic tumors are anesthetized and examined with a miniendoscope. The endoscope is introduced via the anus and the colon is carefully insufflated with an air pump. Endoscopic pictures obtained are of high quality and allow the monitoring and grading of tumors and inflammation. In addition, colonic biopsies can be taken. This protocol can be completed within 1 h. PMID- 17406548 TI - Analysis of the kinetics of folding of proteins and peptides using circular dichroism. AB - Circular dichroism (CD) is a useful spectroscopic technique for studying the secondary structure, folding and binding properties of proteins. This protocol covers how to use the intrinsic circular dichroic properties of proteins to follow their folding and unfolding as a function of time. Included are methods of obtaining data and for analyzing the folding and unfolding data to determine the rate constants and the order of the folding and unfolding reactions. The protocol focuses on the use of CD to follow folding when it is relatively slow, on the order of minutes to days. The methods for analyzing the data, however, can also be applied to data collected with a CD machine equipped with stopped-flow accessories in the range of milliseconds to seconds and folding analyzed by other spectroscopic methods including changes in absorption or fluorescence spectra as a function of time. PMID- 17406550 TI - Assays for in vitro monitoring of proliferation of human airway smooth muscle (ASM) and human pulmonary arterial vascular smooth muscle (VSM) cells. AB - Vascular and airway remodeling, which are characterized by airway smooth muscle (ASM) and pulmonary arterial vascular smooth muscle (VSM) proliferation, contribute to the pathology of asthma, pulmonary hypertension, restenosis and atherosclerosis. To evaluate the proliferation of VSM and ASM cells in response to mitogens, we perform a [3H]thymidine incorporation assay. The proliferation protocol takes approximately 48 h and includes stimulating cells synchronized in G0/G1 phase of the cell cycle with agonists, labeling cells with [3H]thymidine and examining levels of [3H]thymidine incorporation by scintillation counting. Although using radiolabeled [3H]thymidine incorporation is a limitation, the greatest benefit of the assay is providing reliable and statistically significant data. PMID- 17406551 TI - Cyanogen bromide-activated coupling: DNA catalytic chromatography purification of EcoRI endonuclease. AB - A method to purify enzymes utilizing their specific biological affinity and catalytic specificity is described. For this chromatographic technique, an enzyme binds immobilized substrate coupled to a column in the absence of a cofactor required for catalysis but permissive for substrate binding. After washing, the missing cofactor is added to the column mobile phase, and the enzyme converts substrate into product and elutes from the column. A single-step purification of EcoRI endonuclease using a sequence-specific DNA column (containing the GAATTC motif coupled to cyanogen bromide-activated Sepharose 4B) binds EcoRI in the absence of Mg2+ and elutes when Mg2+ is applied in a highly purified state. Although the method described is specific for EcoRI, it can be readily modified for the purification of DNA polymerases and other enzymes. Furthermore, many of the same materials are also used for transcription factor purification. This protocol can be completed within 4-6 d. PMID- 17406552 TI - Imaging synaptic vesicle exocytosis and endocytosis with FM dyes. AB - FM dyes have been used to label and then monitor synaptic vesicles, secretory granules and other endocytic structures in a variety of preparations. Here, we describe the general procedure for using FM dyes to study endosomal trafficking in general, and synaptic vesicle recycling in particular. The dye, dissolved in normal saline solution, is added to a chamber containing the preparation to be labeled. Stimulation evokes exocytosis, and compensatory endocytosis that follows traps FM dye inside the retrieved vesicles. The extracellular dye is then washed from the chamber, and labeled endocytic structures are examined with a fluorescence microscope. Fluorescence intensity provides a direct measure of the labeled vesicle number, a good measure of the amount of exocytosis. If the preparation is stimulated again, without dye in the chamber, dimming of the preparation provides a measure of exocytosis of labeled vesicles. With a synaptic preparation on hand, this protocol requires 1 day. PMID- 17406553 TI - Assays for in vitro monitoring of human airway smooth muscle (ASM) and human pulmonary arterial vascular smooth muscle (VSM) cell migration. AB - Migration of human pulmonary vascular smooth muscle (VSM) cells contributes to vascular remodeling in pulmonary arterial hypertension and atherosclerosis. Evidence also indicates that, in part, migration of airway smooth muscle (ASM) cells may contribute to airway remodeling associated with asthma. Here we describe migration of VSM and ASM cells in vitro using Transwell or Boyden chamber assays. Because dissecting signaling mechanisms regulating cell migration requires molecular approaches, our protocol also describes how to assess migration of transfected VSM and ASM cells. Transwell or Boyden chamber assays can be completed in approximately 8 h and include plating of serum-deprived VSM or ASM cell suspension on membrane precoated with collagen, migration of cells toward chemotactic gradient and visual (Transwell) or digital (Boyden chamber) analysis of membrane. Although the Transwell assay is easy, the Boyden chamber assay requires hands-on experience; however, both assays are reliable cell-based approaches providing valuable information on how chemotactic and inflammatory factors modulate VSM and ASM migration. PMID- 17406554 TI - Application of highly sensitive fluorescent dyes (CyDye DIGE Fluor saturation dyes) to laser microdissection and two-dimensional difference gel electrophoresis (2D-DIGE) for cancer proteomics. AB - Proteome data combined with histopathological information provides important, novel clues for understanding cancer biology and reveals candidates for tumor markers and therapeutic targets. We have established an application of a highly sensitive fluorescent dye (CyDye DIGE Fluor saturation dye), developed for two dimensional difference gel electrophoresis (2D-DIGE), to the labeling of proteins extracted from laser microdissected tissues. The use of the dye dramatically decreases the protein amount and, in turn, the number of cells required for 2D DIGE; the cells obtained from a 1 mm2 area of an 8-12 microm thick tissue section generate up to 5,000 protein spots in a large-format 2D gel. This protocol allows the execution of large-scale proteomics in a more efficient, accurate and reproducible way. The protocol can be used to examine a single sample in 5 d or to examine hundreds of samples in large-scale proteomics. PMID- 17406555 TI - Site-specific incorporation of non-natural amino acids into proteins in mammalian cells with an expanded genetic code. AB - We describe a detailed protocol for incorporating non-natural amino acids, 3-iodo L-tyrosine (IY) and p-benzoyl-L-phenylalanine (pBpa), into proteins in response to the amber codon (the UAG stop codon) in mammalian cells. These amino acids, IY and pBpa, are applicable for structure determination and the analysis of a network of protein-protein interactions, respectively. This method involves (i) the mutagenesis of the gene encoding the protein of interest to create an amber codon at the desired site, (ii) the expression in mammalian cells of the bacterial pair of an amber suppressor tRNA and an aminoacyl-tRNA synthetase specific to IY or pBpa and (iii) the supplementation of the growth medium with these amino acids. The amber mutant gene, together with these bacterial tRNA and synthetase genes, is introduced into mammalian cells. Culturing these cells for 16-40 h allows the expression of the full-length product from the mutant gene, which contains the non-natural amino acid at the introduced amber position. This method is implemented using the conventional tools for molecular biology and treating cultured mammalian cells. This protocol takes 5-6 d for plasmid construction and 3-4 d for incorporating the non-natural amino acids into proteins. PMID- 17406556 TI - Fabrication of cell-containing gel modules to assemble modular tissue-engineered constructs [corrected]. AB - This paper describes a protocol to encapsulate cells in sub-millimeter-sized cylindrical collagen modules and to assemble these modules into a tissue engineered construct within a continuous loop flow circuit. Modules are fabricated by gelling a solution of collagen, that contains suspended cells, within the lumen of a small-bore polyethylene tube. The tubing is then cut into short lengths using an automated cutter and gently vortexed to remove the cell containing collagen modules from the tubing lumen. Modules are then randomly assembled into a modular construct by pipetting a suspension of modules into a larger tube that is positioned within a continuous flow circuit. A range of cylinder aspect ratios are achievable; therefore, this method could potentially be used to create short discs or, alternatively, long threads of soft gels, with or without encapsulated cells, for a variety of tissue-engineering applications. Module fabrication requires 1 d and assembly of a modular construct requires 2 h. PMID- 17406557 TI - Studying vesicle cycling in presynaptic terminals using the genetically encoded probe synaptopHluorin. AB - Genetically encoded fluorescent probes have become indispensable tools in the biological sciences. Studies of synaptic vesicle recycling have been facilitated by a group of GFP-derived probes called pHluorins. These probes exploit changes in pH that accompany exocytosis and recapture of synaptic vesicles. Here we describe how these synaptic tracers can be used in rodent hippocampal neurons to monitor the synaptic vesicle cycle in real time and to obtain mechanistic insights about it. Synapses can be observed in living samples using a wide-field fluorescence microscope and a cooled charge-coupled device camera. A simple specimen chamber allows electrical stimulation of synapses to evoke exocytosis in a precisely controlled manner. We present protocols to measure various parameters of the synaptic vesicle cycle. This technique can be easily adapted to study different classes of synapses from wild-type and mutant mice. Once cultured neurons expressing synaptopHluorin are available, the whole procedure should take about 2 h. PMID- 17406558 TI - Adult mouse hematopoietic stem cells: purification and single-cell assays. AB - Mouse hematopoietic stem cells (HSCs) are the best-studied stem cells because functional assays for mouse HSCs were established earliest and purification techniques for mouse HSCs have progressed furthest. Here we describe our current protocols for the purification of CD34-/lowc-Kit+Sca-1+lineage marker- (CD34-KSL) cells, the HSC population making up approximately 0.005% of bone marrow cells in adult C557BL/6 mice. Purified HSCs have been characterized at cellular and molecular levels. Since clonal analysis is essential for the study of self renewal and lineage commitment in HSCs, here we present our single-cell colony assay and single-cell transplantation procedures. We also introduce our immunostaining procedures for small numbers of HSCs, which are useful for signal transduction analysis. The purification of CD34-KSL cells requires approximately 6 h. Initialization of single-cell culture requires approximately 1 h. Single cell transplantation requires approximately 6 h. Single-cell immunostaining requires approximately 2 d. PMID- 17406559 TI - Whole-body imaging of bacterial infection and antibiotic response. AB - We describe imaging of green fluorescent protein (GFP)-expressing bacteria from outside intact infected animals. This simple, non-intrusive technique can show in great detail the spatial-temporal behavior of the infectious process. The bacteria, expressing the GFP, are sufficiently bright as to be clearly visible from outside the infected animal and recorded with simple equipment. Introduced bacteria can be whole-body imaged in most mouse organs, including the peritoneal cavity, stomach, small intestine, and colon. This imaging technology affords a powerful approach to visualizing the infection process, determining the tissue specificity of infection, the spatial migration of the infectious agents and the response to antimicrobial agents. PMID- 17406560 TI - A simple method for quantitative determination of polysaccharides in fungal cell walls. AB - A simple and reliable method for quantitative determination of cell wall polymers in fungal cell with an s.e.m. of 5% is described. This protocol is based on the hydrolysis by sulfuric acid of beta-glucan, mannan, galactomannan and chitin present at different levels in the wall of yeasts and filamentous fungi into their corresponding monomers glucose, mannose, galactose and glucosamine. The released monosaccharides are subsequently separated and quantified by high performance ionic chromatography coupled to pulse amperometry detection, with a detection limit of 1.0 mug ml(-1). This procedure is well suited to screening a large collection of yeast mutants or to evaluating effects of environmental conditions on cell wall polysaccharide content. This procedure is also applicable to other fungal species, including Schizosaccharomyces pombe, Candida albicans and Aspergillus fumigatus. Results can be obtained in 3 d. PMID- 17406561 TI - Dehydroalanine-containing peptides: preparation from phenylselenocysteine and utility in convergent ligation strategies. AB - This protocol describes the methodology for the synthesis of dehydroalanine (Dha) containing peptides and illustrates their use in convergent ligation strategies for the preparation of peptide conjugates. A nonproteinogenic amino acid, Fmoc-Se phenylselenocysteine (SecPh), can be prepared in high yield over four synthetic steps and be conveniently incorporated into peptides by standard solid-phase peptide synthesis techniques. Globally deprotected peptides containing phenylselenocysteine can be converted to dehydrated peptides following a chemoselective, mild oxidation with hydrogen peroxide or sodium periodate (i.e., the phenylselenocysteine side chain is converted to that of Dha). Dha residues are electrophilic handles for the preparation of glycopeptides, lipopeptides or other peptide conjugates; one such transformation will be outlined here. The preparation of Dha-containing peptides, including the synthesis of SecPh, peptide elongation and oxidative treatment of phenylselenocysteine-containing peptides can be completed by one person in approximately 3-5 weeks. However, once SecPh is in hand, the time required for the preparation of peptides is significantly shorter and comparable to that for any peptide synthesis. PMID- 17406562 TI - Two-dimensional strandness-dependent electrophoresis. AB - Two-dimensional strandness-dependent electrophoresis (2D-SDE) separates nucleic acids in complex samples according to strandness, conformation and length. Under the non-denaturing conditions of the first electrophoretic step, single-stranded DNA, double-stranded DNA and RNA.DNA hybrids of similar length migrate at different rates. The second electrophoretic step is performed under denaturing conditions (7 mol l(-1) urea, 55 degrees C) so that all the molecules are single stranded and separate according to length only. 2D-SDE is useful for revealing important characteristics of complex nucleic acid samples in manipulations such as amplification, renaturation, cDNA synthesis and microarray hybridization. It can also be used to identify mispaired, nicked or damaged fragments in double stranded DNA. The protocol takes approximately 2 h and requires only basic skills, equipment and reagents. PMID- 17406563 TI - Mass spectrometric identification of N-linked glycopeptides using lectin-mediated affinity capture and glycosylation site-specific stable isotope tagging. AB - Protein post-translational modifications (PTMs), such as glycosylation and phosphorylation, are crucial for various signaling and regulatory events, and are therefore an important objective of proteomics research. We describe here a protocol for isotope-coded glycosylation site-specific tagging (IGOT), a method for the large-scale identification of N-linked glycoproteins from complex biological samples. The steps of this approach are: (1) lectin column-mediated affinity capture of glycopeptides generated by protease digestion of protein mixtures; (2) purification of the enriched glycopeptides by hydrophilic interaction chromatography (HIC); (3) peptide-N-glycanase-mediated incorporation of a stable isotope tag, 18O18O, specifically at the N-glycosylation site; and (4) identification of 18O-tagged peptides by liquid chromatography-coupled mass spectrometry (LC/MS)-based proteomics technology. The application of this protocol to the characterization of N-linked glycoproteins from crude extracts of the nematode Caenorhabditis elegans or mouse liver provides a list of hundreds to a thousand glycoproteins and their sites of glycosylation within a week. PMID- 17406564 TI - Contour-clamped homogeneous electric field electrophoresis of Staphylococcus aureus. AB - Contour-clamped homogeneous electric field (CHEF) electrophoresis is a technique of pulsed-field gel electrophoresis that enables the resolution of large fragments of DNA that cannot be resolved by conventional gel electrophoresis. The procedure involves the application of controlled electric fields that change direction at a predetermined angle to samples of DNA that have been embedded in an agarose gel matrix and digested with a restriction endonuclease. Adjustment of the electrophoresis conditions enables the separation of DNA fragments with lengths from 10 kilobases up to 9 megabases in a size-dependent manner in agarose gels. The banding patterns can be used for epidemiological typing, the separated DNA can be immobilized onto a membrane and used for genetic mapping, or individual fragments can be extracted and used for downstream genetic manipulations. The protocol requires specialized equipment and can be completed in a maximum of 7 days. PMID- 17406565 TI - In vitro assay for the quantitation of apoptosis-induced alterations of nuclear envelope permeability. AB - This protocol describes how to perform comparative measurements of the permeability of the nuclear envelope in adherent cells. The plasma membrane is permeabilized at low digitonin concentrations, leaving the nuclear membrane intact. These semi-permeabilized cells are incubated with cytosolic extracts prepared in advance and with a fluorescent reporter molecule whose molecular weight exceeds the size-exclusion limit of the nuclear envelope. Images are taken with a confocal microscope and subsequently analyzed using a custom-made software program that recognizes the nuclei automatically and calculates the mean nuclear fluorescence signal. Here, we measure the increase in nuclear permeability triggered by cytosolic extracts from cells dying by apoptosis. This method can be employed for the study of processes that affect the nucleocytoplasmic distribution of fluorescent molecules in cell populations. The large size of the samples means that subtle fluctuations in nuclear fluorescence can be detected with a high confidence level. Isolation of cell extracts takes 5-6 h, and the preparation and imaging of 15 or so specimens takes 4-5 h. PMID- 17406566 TI - Synthesis of light-activated antisense oligodeoxynucleotide. AB - The activity of a 20-mer antisense oligodeoxynucleotide (asODN) is transiently blocked by attaching a partially complementary sense strand (sODN) via a heterobifunctional photocleavable linker (PL). The asODN-PL-sODN conjugate forms a DNA hairpin-like structure that is considerably more stable than the corresponding asODN/sODN duplex. In conjugate form, the asODN is prevented from hybridizing to exogenous RNA or DNA molecules. Activity is restored after modest exposure to UV light (lambda approximately 365 nm). Here, we provide a detailed procedure for synthesizing photoactive asODNs in good yields. Synthesis, purification and analysis of the light-activated asODN can be completed within 1 2 weeks. PMID- 17406567 TI - Retrovirus-mediated single-cell gene knockout technique in adult newborn neurons in vivo. AB - Single-cell genetic manipulation in an intact brain environment is an informative approach to study molecular mechanism of adult neurogenesis. Here, we describe a protocol for retrovirus-mediated single-cell gene knockout in adult new neurons in vivo. A gene of interest is disrupted in adult floxed mice by a vector based on the Moloney murine leukemia retrovirus, expressing Cre recombinase. High-titer retrovirus is prepared by transfecting plasmids into the HEK293T cells and by concentrating the supernatant containing virus. The retrovirus is stereotaxically injected into the dentate gyrus. Cre recombinase is transduced and expressed in a small fraction of adult new neurons in an intact environment, and the gene knockout is highly efficient within the transduced neurons. Virus preparation takes 7 days, but virus injections take less than 1 h per mouse. By changing the survival time of the mice after the injection, one can analyze the effects on new neurons at different ages. PMID- 17406568 TI - Amplification of 5' end cDNA with 'new RACE'. AB - 'New RACE' (rapid amplification of cDNA ends) PCR is a method for obtaining full length cDNA for mRNA for which only part of the sequence is known. Starting with cellular mRNA, PCR is used to amplify regions between the known parts of the sequence and nonspecific tags at the ends of the cDNA. In 'new RACE', an anchor is ligated to the 5' end of the mRNA before reverse transcription, resulting in the selective production of full-length 5' cDNA ends. Although 'new RACE' can also be used to amplify 3' ends, only the protocol for obtaining 5' ends is presented here. This protocol can be completed in 1-3 days. PMID- 17406569 TI - Radiolabeling of TETA- and CB-TE2A-conjugated peptides with copper-64. AB - The number of radiopharmaceuticals containing copper radionuclides for diagnostic imaging and targeted radiotherapy has grown considerably over the past few decades. This expansion has created the need for protocols allowing for the efficient chelation of 64Cu to peptide-chelator conjugates. Step 1A of this protocol describes a (64)Cu-radiolabeling procedure for 1,4,8,11 tetraazacyclododecane-1,4,8,11-tetraacetic acid (TETA)-conjugated peptides. This reaction is facile and requires the incubation of 64CuCl2 in 0.1 M ammonium acetate buffer with the TETA-peptide for 30 min at room temperature (20-23 degrees C). Step 1B of this protocol describes the radiolabeling procedure for 4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CB-TE2A) conjugated peptides. The CB-TE2A-peptide can be labeled with 64Cu in 0.1 M ammonium acetate buffer in 2 h at 95 degrees C. In both cases, the conjugates can be radiolabeled with 64Cu at greater than 95% purity and with specific activities of 37-111 MBq microg(-1) (1-3 mCi microg(-1)). Both protocols are straightforward and can be completed within 3 h. PMID- 17406570 TI - PET imaging of herpes simplex virus type 1 thymidine kinase (HSV1-tk) or mutant HSV1-sr39tk reporter gene expression in mice and humans using [18F]FHBG. AB - The herpes simplex virus type 1 thymidine kinase (HSV1-tk) positron emission tomography (PET) reporter gene (PRG) or its mutant HSV1-sr39tk are used to investigate intracellular molecular events in cultured cells and to image intracellular molecular events and cell trafficking in living subjects. The expression of these PRGs can be imaged using 18F- or 124I-radiolabeled acycloguanosine or pyrimidine analog PET reporter probes (PRPs). This protocol describes the procedures for imaging HSV1-tk or HSV1-sr39tk PRG expression in living subjects with the acycloguanosine analog 9-4-[18F]fluoro-3 (hydroxymethyl)butyl]guanine ([18F]FHBG). [18F]FHBG is a high-affinity substrate for the HSV1-sr39TK enzyme with relatively low affinity for mammalian TK enzymes, resulting in improved detection sensitivity. Furthermore, [18F]FHBG is approved by the US Food and Drug Administration as an investigational new imaging agent and has been shown to detect HSV1-tk transgene expression in the liver tumors of patients. MicroPET imaging of each small animal can be completed in approximately 1.5 h, and each patient imaging session takes approximately 3 h. PMID- 17406571 TI - Thermal and high pressure intramolecular Diels-Alder reaction of vinylsulfonamides. AB - Vinylsulfonamides with a furan, carbocyclic, semicyclic or acyclic 1,3-diene moiety are synthetized via a domino elimination-amidation reaction of 2 chloroethanesulfonyl chloride. Intramolecular Diels-Alder reaction of these vinylsulfonamides with thermal (toluene, 110 degrees C) or high pressure (dichloromethane, 13 kbar) activation provides efficient access to a range of gamma- and delta-sultams following a 2-3 d long synthetic procedure. Enantiopure sultams are readily obtained from N-1-phenylethyl substituted vinylsulfonamides. PMID- 17406572 TI - A continuous method for the large-scale extraction of plasmid DNA by modified boiling lysis. AB - This protocol describes a streamlined method of plasmid DNA extraction by continual thermal lysis, a modification of the basic boiling lysis technique, to simplify the processing of large volumes of Escherichia coli cultures. Fermented bacteria are harvested using a hollow fiber-membrane module and pre-treated with lysozyme prior to passing through a thermal exchange coil set at 70 degrees C to lyse the cells, and into a juxtaposed cooling coil on ice. The lysed and cooled bacteria are subsequently separated from the lysate by centrifugation and plasmid DNA is precipitated from the supernatant for further purification. The use of peristaltic pumps and two heating coils at constant temperature without the use of centrifugation enable the lysis process to become constant and controllable, providing a flow-through protocol for cell lysis and plasmid DNA extraction. Large volumes of bacterial cultures (20 l) can be processed in 2 h, yielding approximately 100 mg plasmid DNA l(-1) culture, making this an attractive protocol for consistent and large-scale preparation of plasmid DNA. PMID- 17406573 TI - Mutation screening using fluorescence multiplex denaturing gradient gel electrophoresis (FMD): detecting mutations in the BRCA1 gene. AB - Fluorescent multiplex denaturing gradient gel electrophoresis (FMD) is a mutation screening technique designed to detect unknown as well as previously identified mutations. FMD constitutes a recent modification of the standard denaturing gradient gel electrophoresis (DGGE) technique, which combines multiplex PCR amplification of target DNA using fluorescently labeled primers with DGGE separation of the amplicon mixture, allowing immediate identification of sequence variants by wet gel scanning. FMD permits the simultaneous detection of small insertions, deletions and single nucleotide substitutions among multiple DNA fragments (up to 480 fragments) from 96 samples in parallel for each run. It increases output and reduces cost dramatically compared with classical DGGE, without sacrificing sensitivity and accuracy in detecting mutations. This protocol details an accurate, fast, nonradioactive and cost-effective way to screen the BRCA1 gene for mutations with high sensitivity, providing easily interpreted results. It may also be adapted to screen other target genes and/or used in large-scale epidemiological studies. PMID- 17406574 TI - Fusion PCR and gene targeting in Aspergillus nidulans. AB - We describe a rapid method for the production of fusion PCR products that can be used, generally without band purification, to transform Aspergillus nidulans. This technique can be used to replace genes; tag genes with fluorescent moeties or epitope tags; or replace endogenous promoters with regulatable promoters, by introducing an appropriate selective cassette (e.g., fluorescent protein + selectable marker). The relevant genomic fragments and cassette are first amplified separately by PCR using primers that produce overlapping ends. A second PCR using 'nested' primers fuses the fragments into a single molecule with all sequences in the desired order. This procedure allows a cassette to be amplified once, frozen and used subsequently in many fusion PCRs. Transformation of nonhomologous recombination deficient (nkuADelta) strains of A. nidulans with fusion PCR products results in high frequencies of accurate gene targeting. Fusion PCR takes less than 2 d. Protoplast formation and transformation takes less than 1 d. PMID- 17406575 TI - Single-strand conformation polymorphism (SSCP) for the analysis of genetic variation. AB - The accurate analysis of genetic variation has major implications in many areas of biomedical research, including the identification of infectious agents (such as parasites), the diagnosis of infections, and the detection of unknown or known disease-causing mutations. Mutation scanning methods, including PCR-coupled single-strand conformation polymorphism (SSCP), have significant advantages over many other nucleic acid techniques for the accurate analysis of allelic and mutational sequence variation. The present protocol describes the SSCP method of analysis, including all steps from the small-scale isolation of genomic DNA and PCR amplification of target sequences, through to the gel-based separation of amplicons and scanning for mutations by SSCP (either by the analysis of radiolabeled amplicons in mutation detection enhancement (MDE) gels or by non isotopic SSCP using precast GMA gels). The subsequent sequence analysis of polymorphic bands isolated from gels is also detailed. The SSCP protocol can readily detect point mutations for amplicon sizes of up to 450-500 bp, and usually takes 1-2 days to carry out. This user-friendly, low-cost, potentially high-throughput platform has demonstrated the utility to study a wide range of pathogens and diseases, and has the potential to be applied to any gene of any organism. PMID- 17406576 TI - Spectral karyotyping analysis of human and mouse chromosomes. AB - Classical banding methods provide basic information about the identities and structures of chromosomes on the basis of their unique banding patterns. Spectral karyotyping (SKY), and the related multiplex fluorescence in situ hybridization (M-FISH), are chromosome-specific multicolor FISH techniques that augment cytogenetic evaluations of malignant disease by providing additional information and improved characterization of aberrant chromosomes that contain DNA sequences not identifiable using conventional banding methods. SKY is based on cohybridization of combinatorially labeled chromosome-painting probes with unique fluorochrome signatures onto human or mouse metaphase chromosome preparations. Image acquisition and analysis use a specialized imaging system, combining Sagnac interferometer and CCD camera images to reconstruct spectral information at each pixel. Here we present a protocol for SKY analysis using commercially available SkyPaint probes, including procedures for metaphase chromosome preparation, slide pretreatment and probe hybridization and detection. SKY analysis requires approximately 6 d. PMID- 17406577 TI - Programmable reactivity-based one-pot oligosaccharide synthesis. AB - A detailed protocol is described for the application of a programmable one-pot oligosaccharide synthesis methodology to the synthesis of fucosyl GM1. This serves as a general example of the application of this method to the synthesis of any desired oligosaccharide. The method relies on a large database of relative reactivities for differentially protected tolyl thioglycoside donor molecules and a computer program to suggest the best order of addition for assembly of the oligosaccharide in optimal yield and with the fewest operations. The product is a protected form of the desired oligosaccharide isolated in 47% yield, which is then deprotected using standard procedures to provide fucosyl GM1 oligosaccharide (1) in 44% yield. The total time for synthesis of 1 from building blocks 3, 4 and 5 is approximately 4 d, whereas synthesis of the same compound by traditional stepwise procedures would take significantly longer. Protocols for the synthesis of thioglycoside building blocks 3 and 4 are also described. PMID- 17406578 TI - Off-wafer fabrication and surface modification of asymmetric 3D SU-8 microparticles. AB - This protocol details a method of fabricating off-wafer, multilayered, asymmetric microparticles from the epoxy-based photosensitive polymer SU-8. Also included are methods for altering the surface properties of SU-8 and introducing functional groups for further covalent coupling reactions. These techniques may be used to create microparticles for applications in drug delivery and tissue engineering. Microparticle design, fabrication and surface modification can be completed in 2 d. PMID- 17406579 TI - Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. AB - The spectrophotometric/microplate reader assay method for glutathione (GSH) involves oxidation of GSH by the sulfhydryl reagent 5,5'-dithio-bis(2 nitrobenzoic acid) (DTNB) to form the yellow derivative 5'-thio-2-nitrobenzoic acid (TNB), measurable at 412 nm. The glutathione disulfide (GSSG) formed can be recycled to GSH by glutathione reductase in the presence of NADPH. The assay is composed of two parts: the preparation of cell cytosolic/tissue extracts and the detection of total glutathione (GSH and GSSG). The method is simple, convenient, sensitive and accurate. The lowest detection for GSH and GSSG is 0.103 nM in a 96 well plate. This method is rapid and the whole procedure takes no longer than 15 min including reagent preparation. The method can assay GSH in whole blood, plasma, serum, lung lavage fluid, cerebrospinal fluid, urine, tissues and cell extracts and can be extended for drug discovery/pharmacology and toxicology protocols to study the effects of drugs and toxic compounds on glutathione metabolism. PMID- 17406580 TI - Stereotaxic gene delivery in the rodent brain. AB - Stereotaxic surgery has been an invaluable tool in systems neuroscience, applied in many experiments for the creation of site-targeted lesions, injection of anatomical tracers or implantation of electrodes or microdialysis probes. In this protocol, we describe stereotaxic surgery optimized for gene delivery by recombinant adeno-associated viruses and lentiviruses in mice and rats. This method allows the manipulation of gene expression in the rodent brain with excellent spatiotemporal control; essentially any brain region of choice can be targeted and cells (or a subpopulation of cells) in that region can be stably genetically altered at any postnatal developmental stage up to adulthood. Many aspects of the method, its versatility, ease of application and high reproducibility, make it an attractive approach for studying genetic, cellular and circuit functions in the brain. The entire protocol can be completed in 1-2 hours. PMID- 17406581 TI - Establishment of human tumor xenografts in immunodeficient mice. AB - Heterotransplantation of human cancer cells or tumor biopsies into immunodeficient rodents (xenograft models) has, for the past two decades, constituted the major preclinical screen for the development of novel cancer therapeutics. Despite limitations, these models have identified clinically efficacious agents, and remain the 'workhorse' of the pharmaceutical industry. However, if therapeutic approaches to treating tumors according to their molecular characteristics are to be achieved, additional new models of human cancer will be required to represent the genetic diversity that exists within tumor histologies. This protocol details a method for establishing xenografts from primary solid-tumor isolates or cells grown in culture. The procedure relies on immunodeficient mice to provide a host for the establishment of human xenografts. The procedure can be completed in 1-2 h with results being obtained in 1-4 months. PMID- 17406582 TI - Synthesis of 3,3'-bis(sulfonato)-4,4'-bis(chloroacetamido)azobenzene and cysteine cross-linking for photo-control of protein conformation and activity. AB - This protocol describes a procedure for the synthesis of 3,3'-bis(sulfonato)-4,4' bis(chloroacetamido)azobenzene (BSBCA), a water-soluble, thiol-reactive, photo switchable cross-linker. In addition, a protocol is outlined for installing the cross-linker in an intramolecular fashion onto proteins bearing two surface exposed Cys residues. BSBCA is designed to be used as an in vitro activity switch that operates by exerting temporal and reversible photo-control over alpha-helix content within synthetic peptides and recombinant proteins. Synthesis of the cross-linker requires approximately 4.5 d, and cross-linking can be performed in 10-12 h. PMID- 17406583 TI - Isolation of intact vacuoles from Arabidopsis rosette leaf-derived protoplasts. AB - Vacuoles are very prominent compartments within plant cells, and understanding of their function relies on knowledge of their content. Here, we present a simple vacuole purification protocol that was successfully used for large-scale isolation of vacuoles, free of significant contamination from other endomembrane compartments. This method is based on osmotic and thermal disruption of mesophyl derived Arabidopsis protoplasts, followed by a density gradient fractionation of the cellular content. The whole procedure, including protoplast isolation, takes approximately 6 h. PMID- 17406584 TI - In vivo imaging of single axons in the mouse spinal cord. AB - We provide a protocol that describes imaging of single fluorescently labeled axons in the spinal cord of living mice. This method takes advantage of transgenic mouse lines in which the thy1-promoter drives the expression of variants of the green fluorescent protein in a small percentage (less than 1%) of sensory neurons. As a consequence, single axons can be resolved in the surgically exposed dorsal column using wide-field epifluorescence microscopy. This approach allows direct observation of axonal degeneration and regeneration in mouse models of spinal cord pathology for several hours or repetitively over the course of several days. PMID- 17406585 TI - Virus and prokaryote enumeration from planktonic aquatic environments by epifluorescence microscopy with SYBR Green I. AB - Viruses are the most abundant biological entities in aquatic environments, typically exceeding the abundance of bacteria by an order of magnitude. The reliable enumeration of virus-like particles in marine microbiological investigations is a key measurement parameter. Although the size of typical marine viruses (20-200 nm) is too small to permit the resolution of details by light microscopy, such viruses can be visualized by epifluorescence microscopy if stained brightly. This can be achieved using the sensitive DNA dye SYBR Green I (Molecular Probes-Invitrogen). The method relies on simple vacuum filtration to capture viruses on a 0.02-microm aluminum oxide filter, and subsequent staining and mounting to prepare slides. Virus-like particles are brightly stained and easily observed for enumeration, and prokaryotic cells can easily be counted on the same slides. The protocol provides an inexpensive, rapid (30 min) and reliable technique for obtaining counts of viruses and prokaryotes simultaneously. PMID- 17406586 TI - The parallel rod floor test: a measure of ataxia in mice. AB - The parallel rod floor test is a new model of ataxia in mice. It allows the simultaneous measurement of ataxia and locomotor activity. This protocol is designed for researchers examining ethanol-induced motor incoordination in mice, but it should be applicable to other sedative/hypnotic drugs and to testing cerebellar mutant mice or mice with engineered genetic defects. This protocol takes 3 d, with the time per day depending on how many animals are tested. The test allows researchers to quantify differences in motor coordination among genotypes of mice that may differ in locomotor activity. Unlike many other methods for assessing incoordination, the parallel rod floor test yields similar patterns of genetic sensitivity across a range of variant forms of the apparatus. PMID- 17406587 TI - Synthesis of N-succinimidyl 4-guanidinomethyl-3-[*I]iodobenzoate: a radio iodination agent for labeling internalizing proteins and peptides. AB - This protocol describes a detailed procedure for the synthesis of N-succinimidyl 4-guanidinomethyl-3-[*I]iodobenzoate ([*I]SGMIB), an agent useful in the radio iodination of proteins, including monoclonal Abs, and peptides that undergo internalization after receptor or antigen binding. In this procedure, the tin precursor N-succinimidyl 4-[N1,N2-bis(tert-butyloxycarbonyl)guanidinomethyl]-3 (trimethylstannyl)benzoate (Boc-SGMTB, 3) was first radio-iodinated to [*I]Boc SGMIB, a derivative of [*I]SGMIB with the guanidine function protected with Boc groups. Treatment of [*I]Boc-SGMIB with trifluoroacetic acid delivered the final product. The total time for the synthesis and purification of [*I]Boc-SGMIB and its subsequent de-protection is approximately 140 min. PMID- 17406588 TI - Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. AB - Mitochondria participate in key metabolic reactions of the cell and regulate crucial signaling pathways including apoptosis. Although several approaches are available to study mitochondrial function in situ are available, investigating functional mitochondria that have been isolated from different tissues and from cultured cells offers still more unmatched advantages. This protocol illustrates a step-by-step procedure to obtain functional mitochondria with high yield from cells grown in culture, liver and muscle. The isolation procedures described here require 1-2 hours, depending on the source of the organelles. The polarographic analysis can be completed in 1 hour. PMID- 17406589 TI - Piezo-actuated mouse intracytoplasmic sperm injection (ICSI). AB - The mouse is a genetically tractable model organism widely used to study mammalian development and disease. However, mouse metaphase II (mII) oocytes are exquisitely sensitive and intracytoplasmic sperm injection (ICSI) with conventional pipettes generally kills them. This problem can be solved with piezo actuated micromanipulation, in which the piezo-electric effect (crystal deformation in response to an externally applied voltage) propels a microinjection needle tip forward in a precise and rapid movement. Piezo-actuated micromanipulation enhances the penetration of membranes and matrices, and mouse ICSI is a major application. Here we describe a comprehensive, step-by-step mouse piezo ICSI protocol for non-specialists that can be completed in 2-4 h. The protocol is a basic prelude to multiple applications, including nuclear transfer cloning, spermatid injection, blastocyst injection, mII transgenesis, and streamlining micromanipulation in primates and livestock. Moreover, piezo ICSI can be used to obtain offspring from 'dead' (non-motile) sperm, enabling trivial sperm freezing protocols for mouse strain storage and shipment. PMID- 17406590 TI - Synthesis and site-specific incorporation of a simple fluorescent pyrimidine. AB - We describe procedures for the synthesis of a fluorescent pyrimidine analog and its site-specific incorporation into a DNA oligomer. The 5'-protected and 3' activated nucleoside 4 is synthesized in three steps with an overall yield of 40%. Site-specific incorporation into a DNA oligomer occurs with greater than 88% coupling efficiency. This isosteric fluorescent DNA analog can be used to monitor denaturation of DNA duplexes via fluorescence and can positively detect the presence of abasic sites in DNA duplexes. The total time for synthesis of the phosphoramidite 4 is about 75 h, whereas the total time for site-specific incorporation of nucleoside 2 into an oligonucleotide and purification of the corresponding oligonucleotide is about 114 hours. PMID- 17406591 TI - Rapid preparation of the mitotic kinesin Eg5 inhibitor monastrol using controlled microwave-assisted synthesis. AB - We present here a protocol for the synthesis of the dihydropyrimidine (DHPM) derivative monastrol, which is known to be a specific mitotic kinesin Eg5 inhibitor. By applying controlled microwave heating under sealed-vessel conditions, the synthesis via the one-pot three-component Biginelli condensation can be performed in a shorter reaction time (30 min) compared with conventional heating methods that normally require several hours of reflux heating. For the purification of the crude target compound, two different methods are presented. The first protocol includes a simple precipitation/filtration step to provide monastrol in 76% isolated yield and high purity so that no recrystallization step is necessary. This can be ascribed to the microwave heating technology in which less side-product formation is typically one of the advantages. In an alternative purification step, column chromatography is performed, which provides the product in a slightly higher yield (86%). Monastrol synthesis can be conducted in approximately 2 h by employing the precipitation/filtration purification method. PMID- 17406592 TI - The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. AB - The elevated plus maze is a widely used behavioral assay for rodents and it has been validated to assess the anti-anxiety effects of pharmacological agents and steroid hormones, and to define brain regions and mechanisms underlying anxiety related behavior. Briefly, rats or mice are placed at the junction of the four arms of the maze, facing an open arm, and entries/duration in each arm are recorded by a video-tracking system and observer simultaneously for 5 min. Other ethological parameters (i.e., rears, head dips and stretched-attend postures) can also be observed. An increase in open arm activity (duration and/or entries) reflects anti-anxiety behavior. In our laboratory, rats or mice are exposed to the plus maze on one occasion; thus, results can be obtained in 5 min per rodent. PMID- 17406593 TI - In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. AB - The in vitro scratch assay is an easy, low-cost and well-developed method to measure cell migration in vitro. The basic steps involve creating a "scratch" in a cell monolayer, capturing the images at the beginning and at regular intervals during cell migration to close the scratch, and comparing the images to quantify the migration rate of the cells. Compared to other methods, the in vitro scratch assay is particularly suitable for studies on the effects of cell-matrix and cell cell interactions on cell migration, mimic cell migration during wound healing in vivo and are compatible with imaging of live cells during migration to monitor intracellular events if desired. Besides monitoring migration of homogenous cell populations, this method has also been adopted to measure migration of individual cells in the leading edge of the scratch. Not taking into account the time for transfection of cells, in vitro scratch assay per se usually takes from several hours to overnight. PMID- 17406595 TI - Long PCR Product Sequencing (LoPPS): a shotgun-based approach to sequence long PCR products. AB - Here we describe a practical procedure for sequencing long PCR products. The method relies on ultrasonic shearing of PCR products, resulting in fragments 700 1,000 nt long. Termini are subsequently repaired to obtain blunt ends and 3' A overhangs are added before TA cloning. A predetermined number of clones are sequenced using an insert-independent primer to obtain an overlapping contig covering the full length of the PCR product. This method is cost effective and enables the complete sequencing of any large PCR product in a high-throughput format. Processing of amplified DNA requires 3 h handling time prior to the ligation step, and the clone library is available 2 d later. The complete sequence information is obtained approximately 5 d after the PCR step, depending on the sequencing procedure adopted. PMID- 17406594 TI - Solid-phase extraction of N-linked glycopeptides. AB - Protein glycosylation is a common post-translational modification and has been increasingly recognized as one of the most prominent biochemical alterations associated with malignant transformation and tumorigenesis. N-linked glycosylation is prevalent in proteins on the extracellular membrane, and many clinical biomarkers and therapeutic targets are glycoproteins. Here, we describe a protocol for solid-phase extraction of N-linked glycopeptides and subsequent identification of N-linked glycosylation sites (N-glycosites) by tandem mass spectrometry. The method oxidizes the carbohydrates in glycopeptides into aldehydes, which can be immobilized on a solid support. The N-linked glycopeptides are then optionally labeled with a stable isotope using deuterium labeled succinic anhydride and the peptide moieties are released by peptide-N glycosidase. In a single analysis, the method identifies hundreds of N-linked glycoproteins, the site(s) of N-linked glycosylation and the relative quantity of the identified glycopeptides. PMID- 17406596 TI - Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co culture with human liver nonparenchymal cell lines. AB - This protocol describes a co-culture system for the in vitro differentiation of mouse embryonic stem cells into hepatocyte-like cells. Differentiation involves four steps: (i) formation of embryoid bodies (EB), (ii) induction of definitive endoderm from 2-d-old EBs, (iii) induction of hepatic progenitor cells and (iv) maturation into hepatocyte-like cells. Differentiation is completed by 16 d of culture. EBs are formed, and cells can be induced to differentiate into definitive endoderm by culture in Activin A and fibroblast growth factor 2 (FGF 2). Hepatic differentiation and maturation of cells is accomplished by withdrawal of Activin A and FGF-2 and by exposure to liver nonparenchymal cell-derived growth factors, a deleted variant of hepatocyte growth factor (dHGF) and dexamethasone. Approximately 70% of differentiated embryonic stem (ES) cells express albumin and can be recovered by albumin promoter-based cell sorting. The sorted cells produce albumin in culture and metabolize ammonia, lidocaine and diazepam at approximately two-thirds the rate of primary mouse hepatocytes. PMID- 17406597 TI - DNA base mismatch detection with bulky rhodium intercalators: synthesis and applications. AB - This protocol describes the syntheses and applications of two metallointercalators, Rh(bpy)2(chrysi)3+ and Rh(bpy)2(phzi)3+, that target single base mismatches in DNA. The complexes bind mismatched DNA sites specifically and, upon photoactivation, promote strand scission neighboring the mismatch. Owing to their high specificity and sequence context independence, targeting mismatches with these complexes offers an attractive alternative to current mismatch- and SNP-detection methodologies. This protocol also describes the synthesis of these complexes and their use in marking mismatched sites. Irradiation of 32P-labeled duplex DNA with either intercalator followed by denaturing PAGE allows the detection of mismatches in oligonucleotides. The protocol also outlines a method for efficient detection of single nucleotide polymorphisms (SNPs) in larger genes or plasmids. Pooled genes are denatured and re-annealed to form heteroduplexes; they are then incubated with either complex, irradiated and analyzed using capillary electrophoresis to probe for mismatches (SNP sites). The synthesis of the metallointercalators requires approximately 5-7 d. The mismatch- and SNP detection experiments each require approximately 3 d. PMID- 17406598 TI - Cytogenetic harvesting of commonly used tumor cell lines. AB - Tumor cell lines are widely used both as disease models and, increasingly, as genomic resources for the ascertainment of new cancer genes. Cytogenetic analysis remains a major route to uncovering the cancer genome. However, cancer cell lines vary inexplicably in their harvesting preferences, which must, therefore, be determined by trial and error. This article describes harvesting protocols optimized empirically for 550 commonly used, mainly human, cancer cell lines together with evidence-based procedures to assist in determining conditions for unlisted cell lines and subsidiary protocols for cytogenetic analysis using G banding and fluorescence in situ hybridization. PMID- 17406599 TI - A generic protocol for the expression and purification of recombinant proteins in Escherichia coli using a combinatorial His6-maltose binding protein fusion tag. AB - We describe a generic protocol for the overproduction and purification of recombinant proteins in Escherichia coli. The strategy utilizes a dual His6 maltose binding protein (HisMBP) affinity tag that can be removed from the target protein by digestion of the fusion protein at a designed site by tobacco etch virus protease. The MBP moiety serves to enhance the solubility and promote the proper folding of its fusion partners, and the polyhistidine tag facilitates its purification to homogeneity. This protocol is divided into three stages, each of which takes approximately 1 week to complete: (i) construction of a HisMBP fusion vector; (ii) a pilot experiment to assess the yield and solubility of the target protein; and (iii) the large-scale production and purification of the target protein. PMID- 17406600 TI - Reverse transfection on cell arrays for high content screening microscopy. AB - Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfection ready' siRNA arrays, each containing 384 samples, takes in total 7 h. Pre-fabricated siRNA arrays can be used without loss of transfection efficiency at least up to 15 months after printing. Different human cell lines that have been successfully transfected using the protocol are presented here. The present protocol has been applied to two genome-wide siRNA screens addressing mitosis and constitutive protein secretion. PMID- 17406601 TI - Analysis and interpretation of quadratic models of receptive fields. AB - In this protocol, we present a procedure to analyze and visualize models of neuronal input-output functions that have a quadratic, a linear and a constant term, to determine their overall behavior. The suggested interpretations are close to those given by physiological studies of neurons, making the proposed methods particularly suitable for the analysis of receptive fields resulting from physiological measurements or model simulations. PMID- 17406602 TI - Preparation of a copper-based fluorescent probe for nitric oxide and its use in mammalian cultured cells. AB - A procedure for the preparation of a copper(II) complex (CuFL) as a fluorescent nitric oxide (NO) detector is described. The fluorescein-based ligand FL can be synthesized in seven reaction steps (overall yield approximately 20%), typically requiring a total time of 9 days. The CuFL probe allows for the detection of NO produced in mammalian cultured cells. The detailed protocol for the use of CuFL for imaging NO in human neuroblastoma SK-N-SH cells takes a total time of approximately 26 h. This includes plating cells on six-well tissue culture plates or imaging dishes, treatment with CuFL, stimulation of NO synthases and imaging by fluorescence microscopy. PMID- 17406603 TI - Molecular PET imaging of HSV1-tk reporter gene expression using [18F]FEAU. AB - Non-invasive imaging of transgene expression requires the appropriate combination of a reporter gene and a reporter probe. [18F]FEAU positron emission tomography (PET) is used for the assessment of herpes simplex virus type-1 thymidine kinase gene expression. Hybrid AAV phage (termed AAVP) can be adapted to transduce mammalian cells by targeting to a specific receptor. We evaluated a targeted AAVP vector using [18F]FEAU PET. This protocol describes [18F]FEAU production and dosing, micro-PET imaging and image analysis. 2-Deoxy-2-trifluoromethanesulfonyl 1,3,5-tri-O-benzoyl-alpha-D-ribofuranose is radio-fluorinated, converted into its 1-bromo derivative and coupled with protected 5-ethyl uracil. The coupled product is hydrolyzed and purified using HPLC. Tumor-bearing animals targeted with either retroviral or AAVP vectors are anesthetized and injected with [18F]FEAU (0.1 mCi per mouse); this is followed 2 h after injection by imaging on a micro-PET. Production of [18F]FEAU requires approximately 3.5 h from the end of bombardment. PET imaging studies require 2-3 h (depending on the number of animals) after synthesis of [18F]FEAU. PMID- 17406604 TI - Substrate activity screening (SAS): a general procedure for the preparation and screening of a fragment-based non-peptidic protease substrate library for inhibitor discovery. AB - Substrate activity screening (SAS) is a fragment-based method for the rapid development of novel substrates and their conversion into non-peptidic inhibitors of Cys and Ser proteases. The method consists of three steps: (i) a library of N acyl aminocoumarins with diverse, low-molecular-weight N-acyl groups is screened to identify protease substrates using a simple fluorescence-based assay; (ii) the identified N-acyl aminocoumarin substrates are optimized by rapid analog synthesis and evaluation; and (iii) the optimized substrates are converted into inhibitors by direct replacement of the aminocoumarin with known mechanism-based pharmacophores. This protocol describes a general procedure for the solid-phase synthesis of a library of N-acyl aminocoumarin substrates and the screening procedure to identify weak binding substrates. PMID- 17406605 TI - Studying properties of neurotransmitter receptors by non-stationary noise analysis of spontaneous postsynaptic currents and agonist-evoked responses in outside-out patches. AB - Chemical synaptic transmission depends on neurotransmitter-gated ion channels concentrated in the postsynaptic membrane of specialized synaptic contacts. The functional characteristics of these neurotransmitter receptor channels are important for determining the properties of synaptic transmission. Whole-cell recording of postsynaptic currents (PSCs) and outside-out patch recording of transmitter-evoked currents are important tools for estimating the single-channel conductance and the number of receptors contributing to the PSC activated by a single transmitter quantum. When single-channel activity cannot be directly resolved, non-stationary noise analysis is a valuable tool for determining these parameters. Peak-scaled non-stationary noise analysis can be used to compensate for quantal variability in synaptic currents. Here, we present detailed protocols for conventional and peak-scaled non-stationary noise analysis of spontaneous PSCs and responses in outside-out patches. In addition, we include examples of computer code for individual functions used in the different stages of non stationary noise analysis. These analysis procedures require 3-8 h. PMID- 17406606 TI - Preparation of integrin alpha(v)beta3-targeting Ab 38C2 constructs. AB - This protocol describes the preparation of Ab constructs using agents that target cells expressing integrins alpha(v)beta3 and alpha(v)beta5, and the monoclonal aldolase Ab 38C2. The targeting agents are equipped with a diketone or vinylketone linker, and selectively react through the reactive Lys residues in the Ab binding sites to form 38C2 conjugates or chemically programmed 38C2 (i.e., cp38C2). The targeting agent possessing a diketone linker reacts with the Lys residues forming an enaminone derivative. By contrast, the vinylketone linker is used as the corresponding acetone adduct (i.e., a pro-vinylketone linker), and this pro-adapter undergoes a 38C2-catalyzed retro-aldol reaction to produce the vinylketone linker, which forms a Michael-type adduct with the Lys residues. The Ab construct formation is achieved in <1 h for the diketone compounds at ambient temperature, and in 2-16 h using the pro-vinylketone linker at 37 degrees C. The 38C2 constructs are retargeted to cells over-expressing integrins, and are potential candidates for immunotherapy. PMID- 17406607 TI - Labeling, detection and identification of newly synthesized proteomes with bioorthogonal non-canonical amino-acid tagging. AB - A major aim of proteomics is the identification of proteins in a given proteome at a given metabolic state. This protocol describes the step-by-step labeling, purification and detection of newly synthesized proteins in mammalian cells using the non-canonical amino acid azidohomoalanine (AHA). In this method, metabolic labeling of newly synthesized proteins with AHA endows them with the unique chemical functionality of the azide group. In the subsequent click chemistry tagging reaction, azide-labeled proteins are covalently coupled to an alkyne bearing affinity tag. After avidin-based affinity purification and on-resin trypsinization, the resulting peptide mixture is subjected to tandem mass spectrometry for identification. In combination with deuterated leucine-based metabolic colabeling, candidate proteins can be immediately validated. Bioorthogonal non-canonical amino-acid tagging can be combined with any subcellular fractionation, immunopurification or other proteomic method to identify specific subproteomes, thereby reducing sample complexity and enabling the identification of subtle changes in a proteome. This protocol can be completed in 5 days. PMID- 17406608 TI - A protocol for differential display of mRNA expression using either fluorescent or radioactive labeling. AB - Since its invention in the early 1990s, differential display (DD) has become one of the most commonly used techniques for identifying differentially expressed genes at the mRNA level. Unlike other genomic approaches, such as DNA microarrays, DD systematically detects changes in mRNA profiles among multiple samples being compared without the need of any prior knowledge of genomic information of the living organism being studied. Here, we present an optimized DD protocol with a fluorescent digital readout as well as traditional radioactive labeling. The resulting streamlined fluorescent DD process offers an unprecedented accuracy, sensitivity and throughput in comprehensive and quantitative analysis of eukaryotic gene expression. Results usually can be obtained within days using a limited number of primer combinations, but a comprehensive DD screen may take weeks or months to accomplish, depending on gene coverage required and the number of differentially expressed genes present within a biological system being compared. PMID- 17406609 TI - Heterotopic vascularized murine cardiac transplantation to study graft arteriopathy. AB - The development of microsurgical techniques has facilitated the establishment of fully vascularized cardiac transplantation models in small mammals. A particularly useful model that has evolved for the study of cardiac allograft vasculopathy (CAV) is a heterotopic (abdominal) vascularized murine cardiac transplantation model. Using this model has permitted the elucidation of genetic, immune and non-immune factors contributing to the development of this inexorable pathological condition, which compromises half of all human cardiac transplants. This protocol details methods for performing the transplant, histomorphometric assessment of the graft vasculature and functional evaluation of the transplanted heart. In experienced hands, the surgical procedure requires approximately 75 min to complete, and vasculopathy results are obtained at 2 months. This model entails a fully vascularized implantation technique in which the donor ascending aorta and pulmonary artery are sutured end-to-side to the recipient abdominal aorta and inferior vena cava, respectively. As this model reliably reproduces immunological and non-immunological features of CAV, investigators can thoroughly explore contributory mechanisms, diagnostic modalities and therapeutic approaches to its mitigation. PMID- 17406610 TI - A protocol for isolation and culture of human umbilical vein endothelial cells. AB - We describe a protocol for easy isolation and culture of human umbilical vein endothelial cells (HUVECs) to supply every researcher with a method that can be applied in cell biology laboratories with minimum equipment. Endothelial cells (ECs) are isolated from umbilical vein vascular wall by a collagenase treatment, then seeded on fibronectin-coated plates and cultured in a medium with Earles' salts and fetal calf serum (FCS), but without growth factor supplementation, for 7 days in a 37 degrees C-5% CO2 incubator. Cell confluency can be monitored by phase-contrast microscopy; ECs can be characterized using cell surface or intracellular markers and checked for contamination. Various protocols can be applied to HUVECs, from simple harvesting to a particular solubilization of proteins for proteomic analysis. PMID- 17406611 TI - A simple, robust and semi-automated parasite egg isolation protocol. AB - Large-scale parasite quantification is required for improving our understanding of the epidemiology and genetics of host-parasite interactions. We describe a protocol that uses a low-density salt solution for flotation and centrifugation of nematode eggs. Subsequently, sucrose flotation and precipitation are used to obtain clear egg preparations. Most traditional quantification protocols such as the McMaster technique are unsuited for the standardized processing of large numbers of samples and the analysis of large amounts of feces per sample. Consequently, they are suited only for small-scale surveys. Our protocol, which can be used to analyze up to 6 g of feces, results in clear egg preparations that are concentrated in wells of a microtiter plate and that are suited for digital recording and automated counting. Starting from a fecal suspension in the first flotation solution to a digital recording requires approximately 40 min per 24 samples. PMID- 17406612 TI - In vitro screening for molecules that affect virus capsid assembly (and other protein association reactions). AB - Protein self-assembly is critical for numerous biological processes. Yet, assembly is rarely targeted by therapeutic agents, in part because it is hard to identify molecules that interfere with protein-protein interactions. Here, we describe a simple fluorescence-based screen for self-association and its application to the assembly of hepatitis B virus capsids. These data are analyzed to identify kinetic and thermodynamic effects--both of which are critical for the viral lifecycle and for understanding the mechanism of assembly effectors. Suggestions are made for modification of this protocol so that it can be applied to other self-assembling systems. With manual pipetting, setting up a plate takes about 2 h, the initial reading takes 1 h and the end point reading the following day takes about 5 min. PMID- 17406613 TI - Acquisition and voxelwise analysis of multi-subject diffusion data with tract based spatial statistics. AB - There is much interest in using magnetic resonance diffusion imaging to provide information on anatomical connectivity in the brain by measuring the diffusion of water in white matter tracts. Among the measures, the most commonly derived from diffusion data is fractional anisotropy (FA), which quantifies local tract directionality and integrity. Many multi-subject imaging studies are using FA images to localize brain changes related to development, degeneration and disease. In a recent paper, we presented a new approach, tract-based spatial statistics (TBSS), which aims to solve crucial issues of cross-subject data alignment, allowing localized cross-subject statistical analysis. This works by transforming the data from the centers of the tracts that are consistent across a study's subjects into a common space. In this protocol, we describe the MRI data acquisition and analysis protocols required for TBSS studies of localized change in brain connectivity across multiple subjects. PMID- 17406614 TI - The chemoinvasion assay: a method to assess tumor and endothelial cell invasion and its modulation. AB - Invasive and metastatic cells, as well as endothelial cells, must cross basement membranes (BMs) in order to disseminate or to form new blood vessels. The chemoinvasion assay using the reconstituted BM Matrigel in Boyden blind-well chambers is a very rapid, easy, inexpensive and flexible test that can be used to quantify the invasive potential of most cell types; it can be applied to detect the migratory activity associated with matrix degradation and can also be adapted to study the selective degrading activity on different matrix substrates. Transwell inserts can also be used. Once the optimal experimental conditions are empirically determined for specific cellular models, the chemoinvasion assay can be used for the screening of inhibitors of invasiveness and angiogenesis, or to select for invasive cellular populations. This protocol can be completed in 9 h. PMID- 17406615 TI - Immuno-spin trapping analyses of DNA radicals. AB - Immuno-spin trapping is a highly sensitive method for detecting DNA radicals in biological systems. This technique involves three main steps: (i) in situ and real-time trapping of DNA radicals with the nitrone spin trap 5,5-dimethyl-1 pyrroline-N-oxide (DMPO), thus forming DMPO-DNA nitrone adducts (referred to here as nitrone adducts); (ii) purification of nitrone adducts; and (iii) analysis of nitrone adducts by heterogeneous immunoassays using Abs against DMPO. In experiments, DMPO is added prior to the formation of free radicals. It diffuses easily through all cell compartments and is present when DNA free radicals are formed as a result of oxidative damage. Due to its low toxicity, DMPO can be used in cells at high enough concentrations to out-compete the normal reactions of DNA radicals, thus ensuring a high yield of DNA nitrone adducts. Because both protein and DNA nitrone adducts are formed, it is important that the DNA be pure in order to avoid misinterpretations. Depending on the model under study, this protocol can be completed in as few as 6 h. PMID- 17406616 TI - Design and construction of targeted AAVP vectors for mammalian cell transduction. AB - Bacteriophage (phage) evolved as bacterial viruses, but can be adapted to transduce mammalian cells through ligand-directed targeting to a specific receptor. We have recently reported a new generation of hybrid prokaryotic eukaryotic vectors, which are chimeras of genetic cis-elements of recombinant adeno-associated virus and phage (termed AAVP). This protocol describes the design and construction of ligand-directed AAVP vectors, production of AAVP particles and the methodology to transduce mammalian cells in vitro and to target tissues in vivo after systemic administration. Targeted AAVP particles are made in a two-step process. First, a ligand peptide of choice is displayed on the coat protein to generate a targeted backbone phage vector. Then, a recombinant AAV carrying a mammalian transgene cassette is inserted into an intergenomic region. High-titer suspensions (approximately 10(10)-10(11) transducing units per microl) can be produced within 3 days after vector construction. Transgene expression by targeted AAVP usually reaches maximum levels within 1 week. PMID- 17406617 TI - Chemically induced mouse models of intestinal inflammation. AB - Animal models of intestinal inflammation are indispensable for our understanding of the pathogenesis of Crohn disease and ulcerative colitis, the two major forms of inflammatory bowel disease in humans. Here, we provide protocols for establishing murine 2,4,6-trinitro benzene sulfonic acid (TNBS)-, oxazolone- and both acute and chronic dextran sodium sulfate (DSS) colitis, the most widely used chemically induced models of intestinal inflammation. In the former two models, colitis is induced by intrarectal administration of the covalently reactive reagents TNBS/oxazolone, which are believed to induce a T-cell-mediated response against hapten-modified autologous proteins/luminal antigens. In the DSS model, mice are subjected several days to drinking water supplemented with DSS, which seems to be directly toxic to colonic epithelial cells of the basal crypts. The procedures for the hapten models of colitis and acute DSS colitis can be accomplished in about 2 weeks but the protocol for chronic DSS colitis takes about 2 months. PMID- 17406618 TI - Freeze-fracture electron microscopy. AB - The freeze-fracture technique consists of physically breaking apart (fracturing) a frozen biological sample; structural detail exposed by the fracture plane is then visualized by vacuum-deposition of platinum-carbon to make a replica for examination in the transmission electron microscope. The four key steps in making a freeze-fracture replica are (i) rapid freezing, (ii) fracturing, (iii) replication and (iv) replica cleaning. In routine protocols, a pretreatment step is carried out before freezing, typically comprising fixation in glutaraldehyde followed by cryoprotection with glycerol. An optional etching step, involving vacuum sublimation of ice, may be carried out after fracturing. Freeze fracture is unique among electron microscopic techniques in providing planar views of the internal organization of membranes. Deep etching of ultrarapidly frozen samples permits visualization of the surface structure of cells and their components. Images provided by freeze fracture and related techniques have profoundly shaped our understanding of the functional morphology of the cell. PMID- 17406619 TI - Construction and use of spotted large-insert clone DNA microarrays for the detection of genomic copy number changes. AB - Microarray-based comparative genomic hybridization has become a widespread method for the analysis of DNA copy number changes across the human genome. Initial methods for microarray construction using large-insert clones required the preparation of DNA from large-scale cultures. This rapidly became an expensive and time-consuming process when expanded to the number of clones needed for higher resolution arrays. To overcome this problem, several PCR-based strategies have been developed to enable array construction from small amounts of cloned DNA. Here, we describe the construction of microarrays composed of human-specific large-insert clones (40-200 kb) using a specific degenerate oligonucleotide PCR strategy. In addition, we also describe array hybridization using manual and automated procedures and methods for array analysis. The technology and protocols described in this article can easily be adapted for other species dependent on the availability of clone libraries. According to our protocols, the procedure will take approximately 3 days from labeling the DNA to scanning the hybridized slides. PMID- 17406620 TI - Data analysis of assorted serum peptidome profiles. AB - Discovery of biomarker patterns using proteomic techniques requires examination of large numbers of patient and control samples, followed by data mining of the molecular read-outs (e.g., mass spectra). Adequate signal processing and statistical analysis are critical for successful extraction of markers from these data sets. The protocol, specifically designed for use in conjunction with MALDI TOF-MS-based serum peptide profiling, is a data analysis pipeline, starting with transfer of raw spectra that are interpreted using signal processing algorithms to define suitable features (i.e., peptides). We describe an algorithm for minimal entropy-based peak alignment across samples. Peak lists obtained in this way, and containing all samples, all peptide features and their normalized MS-ion intensities, can be evaluated, and results validated, using common statistical methods. We recommend visual inspection of the spectra to confirm all results, and have written freely available software for viewing and color-coding of spectral overlays. PMID- 17406621 TI - Sequencing complete mitochondrial and plastid genomes. AB - Organelle genomics has become an increasingly important research field, with applications in molecular modeling, phylogeny, taxonomy, population genetics and biodiversity. Typically, research projects involve the determination and comparative analysis of complete mitochondrial and plastid genome sequences, either from closely related species or from a taxonomically broad range of organisms. Here, we describe two alternative organelle genome sequencing protocols. The "random genome sequencing" protocol is suited for the large majority of organelle genomes irrespective of their size. It involves DNA fragmentation by shearing (nebulization) and blunt-end cloning of the resulting fragments into pUC or BlueScript-type vectors. This protocol excels in randomness of clone libraries as well as in time and cost-effectiveness. The "long-PCR-based genome sequencing" protocol is specifically adapted for DNAs of low purity and quantity, and is particularly effective for small organelle genomes. Library construction by either protocol can be completed within 1 week. PMID- 17406622 TI - Synthesis and oligonucleotide incorporation of fluorescent cytosine analogue tC: a promising nucleic acid probe. AB - The tricyclic cytosine, tC, is a fluorescent base analogue with excellent properties for investigating intrinsic characteristics of nucleic acid as well as interactions between nucleic acids and other molecules. Its unique fluorescence properties and insignificant influence on overall structure and dynamics of nucleic acid after incorporation makes tC particularly interesting in fluorescence resonance energy transfer and anisotropy measurements. We here describe a straightforward synthesis of the standard monomer form of tC for DNA solid-phase synthesis, the tC phosphoramidite, and its subsequent incorporation into oligonucleotides. The total synthesis of the tC phosphoramidite takes approximately 8 days and its incorporation and the subsequent oligonucleotide purification an additional day. PMID- 17406623 TI - Parallel synthesis of peptide libraries using microwave irradiation. AB - The application of microwave irradiation to solid-phase peptide synthesis increases product purity and reduces reaction time. Parallel synthesis in 96-well polypropylene filter plates with microwave irradiation is an efficient method for the rapid generation of combinatorial peptide libraries in sufficient purity to assay the products directly for biological activity without HPLC purification. In this protocol, the solid-phase support is arrayed into each well of a 96-well plate, reagents are delivered using a multichannel pipette and a microwave reactor is used to complete peptide coupling reactions in 6 min and Fmoc-removal reactions in 4 min under temperature-controlled conditions. The microwave assisted parallel peptide synthesis protocol has been used to generate a library of difficult hexa-beta-peptides in 61% average initial purity (50% yield) and has been applied to the preparation of longer alpha- and beta-peptides. Using this protocol, a library of 96 different hexapeptides can be synthesized in 24 h (excluding characterization). PMID- 17406624 TI - The use of the Ugi four-component condensation. AB - This protocol describes a procedure for the Ugi four-component condensation. It describes the general mechanism as well as the effects of the nature of the components on the Ugi reaction. It also describes the effects of the reaction conditions on the reaction, along with special procedures and workup. The experimental procedure is exemplified by a description of the preparation of N cyclohexyl 2-[N-(2-chloroacetyl)-N-(4-chlorobenzyl)]amino-2-(4 chlorophenyl)acetamide, a typical Ugi product, that is subsequently used for the synthesis of a 2,5-diketopiperazine, an example of an important type of pharmaceutical compound. The experimental procedure is then extended to the synthesis of a 1,5-disubstituted tetrazole via Ugi four-component condensation. The protocol describes the preparation and characterization of the new 1 cyclohexyl-5-(1-phenylamino-2-methyl)propyltetrazole. The total time for the synthesis and isolation of the two example reactions in parallel is 3 d. PMID- 17406625 TI - Selenium modification of nucleic acids: preparation of phosphoroselenoate derivatives for crystallographic phasing of nucleic acid structures. AB - This protocol describes a simplified means of introducing an anomalously scattering atom into oligonucleotides by conventional solid-phase synthesis. Replacement of a nonbridging phosphate oxygen in the backbone with selenium is practically suitable for any nucleic acid. The resulting oligonucleotide P diastereomers can be separated using anion exchange HPLC to yield diastereomerically pure phosphoroselenoates (PSes). The total time for the synthesis and ion-exchange HPLC separation of pure PSe is approximately 60 h. PMID- 17406626 TI - Selenium modification of nucleic acids: preparation of oligonucleotides with incorporated 2'-SeMe-uridine for crystallographic phasing of nucleic acid structures. AB - This protocol describes a method for introducing an anomalously scattering atom into oligonucleotides at the 2'-position of uridine by conventional solid-phase synthesis. The 2'-SeMe ribose modification is particularly attractive for derivatization of RNA to facilitate crystal structure determination. The estimated time for the synthesis and HPLC purification of oligonucleotides with incorporated 2'-SeMe-uridine residues is approximately 46 h for 'trityl on' and approximately 32 h for 'trityl off' methods, respectively. PMID- 17406627 TI - Purification of mitochondrial and plastid DNA. AB - The size, structure and conformation of mitochondrial and plastid genomes differ dramatically among eukaryotes. Similarly, the yield and purity of extracted organelle DNA also vary, and are crucial factors for the success of restriction mapping and sequencing experiments. We describe here procedures for the purification of organelle DNA from a broad range of eukaryotes. By emphasizing the underlying principles, these procedures will facilitate the development of new species-specific protocols. The presented purification schemes involve either isolation of organelles and subsequent extraction of DNA from this subcellular fraction, or processing of whole-cell lysates followed by CsCl gradient centrifugation to separate nuclear and organelle DNAs according to their A + T content. We have successfully used the described procedures for organelle genome sequencing from diverse eukaryotes, including non-axenic protists. Procedures can be completed in 3-5 days, typically yielding a few micrograms of DNA-ample for sequencing complete genomes. PMID- 17406628 TI - Non-invasive measurement of bioelectric currents with a vibrating probe. AB - Small d.c. electrical signals have been detected in many biological systems and often serve important functions in cells and organs. For example, we have recently found that they play a far more important role in directing cell migration in wound healing than previously thought. Here, we describe the manufacture and use of a simplified ultrasensitive vibrating probe system for measuring extracellular electrical currents. This vibrating probe is an insulated, sharpened metal wire with a small platinum-black tip (10-30 microm), which can detect ionic currents in the microA cm(-2) range in physiological saline. The probe is vibrated at about 300 Hz by a piezoelectric bender. In the presence of an ionic current, the probe detects a voltage difference between the extremes of its movement. The basic, low-cost system we describe is readily adaptable to most laboratories interested in measuring physiological electric currents associated with wounds, developing embryos and other biological systems. PMID- 17406629 TI - Mixed-culture assays for analyzing neuronal synapse formation. AB - The assembly of synapses in the vertebrate central nervous system requires bidirectional signaling across the synaptic cleft that directs the differentiation of pre- and postsynaptic membrane domains. Biochemical and genetic studies have identified several adhesion and signaling molecules that localize to synapses and might participate in organizing synaptic structures. Understanding how individual proteins contribute to synaptic organization is complicated by the fact that there are significant numbers of separate signals that cooperate in this process. This protocol describes an assay system that permits examination of synaptogenic activities of individual cell-surface proteins in isolation. Besides the time needed for preparation and growth of primary neuronal cultures (6-14 days), the execution and analysis of the assay is rapid, requiring approximately 2 days. Using this assay, recent studies revealed that single synaptic adhesion complexes can direct a remarkable degree of synaptic differentiation and provided new insights into the cell biological mechanisms of synaptogenesis. PMID- 17406630 TI - Pulsed-field gel electrophoresis. AB - This protocol describes pulsed-field gel electrophoresis (PFGE), a method developed for separation of large DNA molecules. Whereas standard DNA gel electrophoresis commonly resolves fragments up to approximately 50 kb in size, PFGE fractionates DNA molecules up to 10 Mb. The mechanism driving these separations exploits the fact that very large DNA molecules unravel and "snake" through a gel matrix, and such electrophoretic trajectories are perturbed in a size-dependent manner by carefully oriented electrical pulses. PFGE has enabled the rapid genomic analysis of microbes and mammalian cells, and motivated development of large-insert cloning systems such as bacterial and yeast artificial chromosomes. As such, this protocol includes descriptions of two types of PFGE instrumentation (not commercially available), along with detailed instructions for their operation. Additionally, this protocol provides basic instructions for the preparation of intact chromosomal DNA from several types of organisms. PFGE takes 2-3 days, excluding sample preparation. PMID- 17406631 TI - MetaNetwork: a computational protocol for the genetic study of metabolic networks. AB - We here describe the MetaNetwork protocol to reconstruct metabolic networks using metabolite abundance data from segregating populations. MetaNetwork maps metabolite quantitative trait loci (mQTLs) underlying variation in metabolite abundance in individuals of a segregating population using a two-part model to account for the often observed spike in the distribution of metabolite abundance data. MetaNetwork predicts and visualizes potential associations between metabolites using correlations of mQTL profiles, rather than of abundance profiles. Simulation and permutation procedures are used to assess statistical significance. Analysis of about 20 metabolite mass peaks from a mass spectrometer takes a few minutes on a desktop computer. Analysis of 2,000 mass peaks will take up to 4 days. In addition, MetaNetwork is able to integrate high-throughput data from subsequent metabolomics, transcriptomics and proteomics experiments in conjunction with traditional phenotypic data. This way MetaNetwork will contribute to a better integration of such data into systems biology. PMID- 17406632 TI - Uptake and release protocol for assessing membrane binding and permeation by way of isothermal titration calorimetry. AB - The activity of many biomolecules and drugs crucially depends on whether they bind to biological membranes and whether they translocate to the opposite lipid leaflet and trans aqueous compartment. A general strategy to measure membrane binding and permeation is the uptake and release assay, which compares two apparent equilibrium situations established either by the addition or by the extraction of the solute of interest. Only solutes that permeate the membrane sufficiently fast do not show any dependence on the history of sample preparation. This strategy can be pursued for virtually all membrane-binding solutes, using any method suitable for detecting binding. Here, we present in detail one example that is particularly well developed, namely the nonspecific membrane partitioning and flip-flop of small, nonionic solutes as characterized by isothermal titration calorimetry. A complete set of experiments, including all sample preparation procedures, can typically be accomplished within 2 days. Analogous protocols for studying charged solutes, virtually water-insoluble, hydrophobic compounds or specific ligands are also considered. PMID- 17406633 TI - A protocol describing pharynx counts and a review of other assays of apoptotic cell death in the nematode worm Caenorhabditis elegans. AB - Studies of the nematode worm Caenorhabditis elegans have provided important insights into the genetics of programmed cell death (PCD), and revealed molecular mechanisms conserved from nematodes to humans. The organism continues to offer opportunities to investigate the processes of apoptosis under very well-defined conditions and at single-cell resolution in living animals. Here, a survey of the common methods used to study the process of PCD in C. elegans is described. Detailed instructions are provided for one standard method--the counting of extra cells of the anterior pharynx--a quantitative technique that can be used to detect even very subtle alterations in the progression of apoptotic cell death. PMID- 17406634 TI - Determining the stoichiometry and interactions of macromolecular assemblies from mass spectrometry. AB - The growing number of applications to determine the stoichiometry, interactions and even subunit architecture of protein complexes from mass spectra suggests that some general guidelines can now be proposed. In this protocol, we describe the necessary steps required to maintain interactions between subunits in the gas phase. We begin with the preparation of suitable solutions for electrospray (ES) and then consider the transmission of complexes through the various stages of the mass spectrometer until their detection. Subsequent steps are also described, including the dissociation of these complexes into multiple subcomplexes for generation of interaction networks. Throughout we highlight the critical experimental factors that determine success. Overall, we develop a generic protocol that can be carried out using commercially available ES mass spectrometers without extensive modification. PMID- 17406635 TI - Quantitative prediction of cellular metabolism with constraint-based models: the COBRA Toolbox. AB - The manner in which microorganisms utilize their metabolic processes can be predicted using constraint-based analysis of genome-scale metabolic networks. Herein, we present the constraint-based reconstruction and analysis toolbox, a software package running in the Matlab environment, which allows for quantitative prediction of cellular behavior using a constraint-based approach. Specifically, this software allows predictive computations of both steady-state and dynamic optimal growth behavior, the effects of gene deletions, comprehensive robustness analyses, sampling the range of possible cellular metabolic states and the determination of network modules. Functions enabling these calculations are included in the toolbox, allowing a user to input a genome-scale metabolic model distributed in Systems Biology Markup Language format and perform these calculations with just a few lines of code. The results are predictions of cellular behavior that have been verified as accurate in a growing body of research. After software installation, calculation time is minimal, allowing the user to focus on the interpretation of the computational results. PMID- 17406636 TI - Global single-cell cDNA amplification to provide a template for representative high-density oligonucleotide microarray analysis. AB - We describe here a protocol for the representative amplification of global mRNAs from typical single mammalian cells to provide a template for high-density oligonucleotide microarray analysis. A single cell is lysed in a tube without purification and first-strand cDNAs are synthesized using a poly(dT)-tailed primer. Unreacted primer is specifically eliminated by exonuclease treatment and second strands are generated with a second poly(dT)-tailed primer after poly(dA) tailing of the first-strand cDNAs. The cDNAs are split into four tubes, which are independently directionally amplified by PCR, and then recombined. The amplified products (approximately 100 ng) show superior representation and reproducibility of original gene expression, especially for genes expressed in more than 20 copies per cell, compared with those obtained by a conventional PCR protocol, and can effectively be used for quantitative PCR and EST analyses. The cDNAs are then subjected to another PCR amplification with primers bearing the T7 promoter sequence. The resultant cDNA products are gel purified, amplified by one final cycle and used for isothermal linear amplification by T7 RNA polymerase to synthesize cRNAs for microarray hybridization. This protocol yields cDNA templates sufficient for more than 80 microarray hybridizations from a single cell, and can be completed in 5-6 days. PMID- 17406637 TI - A real-time method of imaging glucose uptake in single, living mammalian cells. AB - This protocol details a method for monitoring glucose uptake into single, living mammalian cells using a fluorescent D-glucose derivative, 2-[N-(7-nitrobenz-2-oxa 1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), as a tracer. The specifically designed chamber and superfusion system for evaluating 2-NBDG uptake into cells in real time can be combined with other fluorescent methods such as Ca2+ imaging and the subsequent immunofluorescent classification of cells exhibiting divergent 2-NBDG uptake. The whole protocol, including immunocytochemistry, can be completed within 2 d (except for cell culture). The procedure for 2-NBDG synthesis is also presented. PMID- 17406639 TI - Mutation analysis of candidate genes within the 2q33.3 linkage area for familial early-onset generalised osteoarthritis. AB - In a genome-wide linkage scan of seven families with familial early-onset osteoarthritis (FOA), we mapped a FOA locus to a 5 cM region on chromosome 2q33.3 2q34 with a maximum LOD score of 6.05. To identify causal variants, 17 positional candidate genes and FRZB were sequenced for coding, splice sites, and 5' and 3' untranslated regions. The pathogenicity of possible disease-causing variants was evaluated using predicted effects on protein structure and function, splicing enhancers, degree of conservation and frequency in 790 unrelated subjects from the population-based Rotterdam study scored for the presence of radiographic signs of OA (ROA). Nine novel variants, identified in NRP2, XM_371590, ADAM23, IDH1, PIP5K3 and PTHR2, cosegregated with FOA, of which two were promising. IDH1 Y183C cosegregated in one family, involved a conserved amino-acid change and showed a damaging effect predicted by PolyPhen and SIFT. In the Rotterdam sample, carriers of IDH1 Y183C (0.02) had an increased but insignificant risk for generalised ROA. The second variant, NRP2 c.1938-21T>C cosegregated in three families. In the Rotterdam sample, carriers conferred an increased risk of 2.1 (95% confidence interval, 1.1-4.1, P=0.032) to have generalised ROA. Furthermore, two variants (NRP2 c.1938-21T>C and IDH1 c.933-28C>T) occurred together on the haplotypes segregating with FOA in two out seven families. This haplotype was rare in the Rotterdam sample (0.0013). Two promising variants in or near NRP2 and IDH1 may not be sufficient for the onset of FOA alone but might have a modulating role with FOA. Confirmation in other OA populations is required. PMID- 17406640 TI - Epidemiology and penetrance of Leber hereditary optic neuropathy in Finland. AB - We have performed an entire-population-based survey of the epidemiology and penetrance of Leber hereditary optic neuropathy (LHON) in Finland - a country that is among the best-studied genetic isolates in the world. During our long term clinical follow-up period since 1970, we have so far identified 36 LHON families in Finland, comprised of almost 1000 family members. Counting the unaffected family members has been possible thanks to accessible genealogical records, and this has improved the accuracy of our penetrance figures by minimizing the sample bias. Our results, although confirming some well-known features of LHON, indicate that the overall penetrance of LHON is lower than previously estimated, and that affected females have a higher incidence of affected offspring compared to the unaffected females. The prevalence of LHON in Finland is 1:50 000, and one in 9000 Finns is a carrier of one of the three LHON primary mutations. PMID- 17406641 TI - Specific combinations of HLA-DR2 and DR3 class II haplotypes contribute graded risk for disease susceptibility and autoantibodies in human SLE. AB - The human leukocyte antigen (HLA) Class II antigen presentation alleles DR and DQ are associated with susceptibility to systemic lupus erythematosus (SLE) and the production of lupus-related autoantibodies. Here, we explore the effect of different combinations of Class II risk haplotypes in a large, multi-center collection of 780 SLE families. Haplotypes bearing the DRB1*1501/DQB1*0602 (DR2) and DRB1*0301/DQB1*0201 (DR3) alleles were present in nearly two-thirds of SLE cases and were significantly associated with disease susceptibility in both family-based and case-control study designs. DR3-containing haplotypes conferred higher risk for disease than DR2, and individual homozygous for DR3 or compound heterozygous for DR3 and DR2 showed the highest risk profile. DR2 haplotypes were also found to be associated with antibodies to the nuclear antigen Sm, and, as previously observed, DR3 genotypes were associated with Ro and La autoantibodies. Interestingly, SLE cases and unaffected family members who were DR2/DR3 compound heterozygotes showed particularly strong risk of developing antibodies to Ro, and were enriched for La and Sm. These data provide convincing evidence that particular combinations of HLA Class II DR2 and DR3 haplotypes are key determinants of autoantibody production and disease susceptibility in human SLE. PMID- 17406642 TI - Inherited PAX6, NF1 and OTX2 mutations in a child with microphthalmia and aniridia. AB - A girl with aniridia, microphthalmia, microcephaly and cafe au lait macules was found to have mutations in PAX6, NF1 and OTX2. A novel PAX6 missense mutation (p.R38W) was inherited from her mother whose iris phenotype had not been evident because of ocular neurofibromatosis. Analysis of the NF1 gene in the proband, prompted by the mother's diagnosis and the presence of cafe au lait spots, revealed a nonsense mutation (p.R192X). Subsequently an OTX2 nonsense mutation (p.Y179X) was identified and shown to be inherited from her father who was initially diagnosed with Leber's congenital amaurosis. Since individual mutations in PAX6, OTX2 or NF1 can cause a variety of severe developmental defects, the proband's phenotype is surprisingly mild. This case shows that patients with complex phenotypes should not be eliminated from subsequent mutation analysis after one or even two mutations are found. PMID- 17406643 TI - Transmission ratio distortion and maternal effects confound the analysis of modulators of cystic fibrosis disease severity on 19q13. AB - Two entities localised within in a 5 Mb interval on 19q13, that is the transforming growth factor beta 1 (TGFbeta1) and the cystic fibrosis modifier 1, have been reported to modulate disease severity of cystic fibrosis (CF), albeit the designation of the risk allele for TGFbeta1 differs between studies. We have analysed genotyping data at seven microsatellite loci and four single nucleotide polymorphisms targeting the 19q13 area from 37 nuclear CF families with two affected offspring exhibiting extreme clinical phenotypes for indicators of transmission-ration distortion, maternal genetic or maternal non-genetic effects. Evidence for a transmission-ratio distortion was obtained at D19S112 (P=0.0304) near the recently characterised myotonic dystrophy locus myotonic dystrophy protein kinase (DMPK). Maternal and paternal genotype distributions were significantly different at rs1982073 (Leu10Pro at TGFbeta1) whereby all CF sibs heterozygous at rs1982073 inherited the Leu10 allele from their mother (P=0.000132) in our sibling panel. To ask whether the improved survival in CF over the last decades has any influence on TGFbeta1 allele frequencies, we analysed unrelated F508del homozygotes who were stratified by birth cohort. Sensitivity with respect to the survivor bias was reflected by significantly higher incidence of mild cystic fibrosis transmembrane conductance regulator mutation genotypes in the early born patient cohort (P=0.0169), and an allelic imbalance was also observed at TGFbeta1 (P=0.0664). In conclusion, the role of TGFbeta1 as a CF modulator, suggested from studies with a case-control setting, needs to be interpreted with caution unless family-based analysis is carried out to identify parental genetic and non-genetic effects. PMID- 17406644 TI - Glycerol kinase deficiency alters expression of genes involved in lipid metabolism, carbohydrate metabolism, and insulin signaling. AB - Glycerol kinase (GK) is at the interface of fat and carbohydrate metabolism and has been implicated in insulin resistance and type 2 diabetes mellitus. To define GK's role in insulin resistance, we examined gene expression in brown adipose tissue in a glycerol kinase knockout (KO) mouse model using microarray analysis. Global gene expression profiles of KO mice were distinct from wild type with 668 differentially expressed genes. These include genes involved in lipid metabolism, carbohydrate metabolism, insulin signaling, and insulin resistance. Real-time polymerase chain reaction analysis confirmed the differential expression of selected genes involved in lipid and carbohydrate metabolism. PathwayAssist analysis confirmed direct and indirect connections between glycerol kinase and genes in lipid metabolism, carbohydrate metabolism, insulin signaling, and insulin resistance. Network component analysis (NCA) showed that the transcription factors (TFs) PPAR-gamma, SREBP-1, SREBP-2, STAT3, STAT5, SP1, CEBPalpha, CREB, GR and PPAR-alpha have altered activity in the KO mice. NCA also revealed the individual contribution of these TFs on the expression of genes altered in the microarray data. This study elucidates the complex network of glycerol kinase and further confirms a possible role for glycerol kinase deficiency, a simple Mendelian disorder, in insulin resistance, and type 2 diabetes mellitus, a common complex genetic disorder. PMID- 17406645 TI - Drugs of abuse and the aging brain. AB - Substance abuse among older adults has received little attention in the past, presumably because this population has traditionally accounted for only a small percentage of the drug abuse problem in the United States. The aging of the baby boomer generation (born 1946-1964), however, will soon swell the ranks of older adults and dramatically alter the demography of American society. Several observations suggest that this expansion will likely be accompanied by a precipitous increase in the abuse of drugs, including prescription medications and illicit substances, among older adults. While it is now evident that the brain changes continuously across life, how drugs of abuse interact with these age-related changes remains unclear. The dynamic nature of brain function, however, suggests that substance abuse during older age may augment the risks and require unique considerations for diagnosis and treatment. In addition to describing current and projected prevalence estimates of substance abuse among older adults, the present review discusses how aging affects brain systems involved in drug abuse, and explores the potential impact of drug abuse on the aging brain. Future directions for substance abuse research among older adults will also be considered. PMID- 17406646 TI - 5-HTTLPR biases amygdala activity in response to masked facial expressions in major depression. AB - The amygdala is a key structure in a limbic circuit involved in the rapid and unconscious processing of facial emotions. Increased amygdala reactivity has been discussed in the context of major depression. Recent studies reported that amygdala activity during conscious emotion processing is modulated by a functional polymorphism in the serotonin transporter gene (5-HTTLPR) in healthy subjects. In the present study, amygdala reactivity to displays of emotional faces was measured by means of fMRI at 3T in 35 patients with major depression and 32 healthy controls. Conscious awareness of the emotional stimuli was prevented via backward-masking to investigate automatic emotion processing. All subjects were genotyped for the 5-HTTLPR polymorphism. Risk allele carriers (S or L(G)) demonstrated increased amygdala reactivity to masked emotional faces, which in turn was significantly correlated with life-time psychiatric hospitalization as an index of chronicity. This might indicate that genetic variations of the serotonin transporter could increase the risk for depression chronification via altering limbic neural activity on a preattentive level of emotion processing. PMID- 17406647 TI - Chronic unpredictable stress induces a cognitive deficit and anxiety-like behavior in rats that is prevented by chronic antidepressant drug treatment. AB - Chronic stress is a risk factor for the development of many psychopathological conditions in humans, including major depression and anxiety disorders. There is a high degree of comorbidity of depression and anxiety. Moreover, cognitive impairments associated with frontal lobe dysfunction, including deficits in cognitive set-shifting and behavioral flexibility, are increasingly recognized as major components of depression, anxiety disorders, and other stress-related psychiatric illnesses. To begin to understand the neurobiological mechanisms underlying the cognitive and emotional consequences of chronic stress, it is necessary to employ an animal model that exhibits similar effects. In the present study, a rat model of chronic unpredictable stress (CUS) consistently induced a cognitive impairment in extradimensional set shifting capability in an attentional set shifting test, suggesting an alteration in function of the medial prefrontal cortex. CUS also increased anxiety-like behavior on the elevated plus maze. Further, chronic treatment both with the selective norepinephrine reuptake blocker, desipramine (7.5 mg/kg/day), and the selective serotonin reuptake blocker, escitalopram (10 mg/kg/day), beginning 1 week before CUS treatment and continuing through the behavioral testing period, prevented the CUS-induced deficit in extradimensional set-shifting. Chronic desipramine treatment also prevented the CUS-induced increase in anxiety-like behavioral reactivity on the plus-maze, but escitalopram was less effective on this measure. Thus, CUS induced both cognitive and emotional disturbances that are similar to components of major depression and anxiety disorders. These effects were prevented by chronic treatment with antidepressant drugs, consistent also with clinical evidence that relapse of depressive episodes can be prevented by antidepressant drug treatment. PMID- 17406648 TI - Molecular genetics of the platelet serotonin system in first-degree relatives of patients with autism. AB - Elevated platelet serotonin (5-hydroxytryptamine, 5-HT) is found in a subset of children with autism and in some of their first-degree relatives. Indices of the platelet serotonin system, including whole blood 5-HT, 5-HT binding affinity for the serotonin transporter (K(m)), 5-HT uptake (V(max)), and lysergic acid diethylamide (LSD) receptor binding, were previously studied in 24 first-degree relatives of probands with autism, half of whom were selected for elevated whole blood 5-HT levels. All subjects were then genotyped for selected polymorphisms at the SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 loci. Previous studies allowed an a priori prediction of SLC6A4 haplotypes that separated the subjects into three groups that showed significantly different 5-HT binding affinity (K(m), p=0.005) and 5-HT uptake rate (V(max), p=0.046). Genotypes at four individual polymorphisms in SLC6A4 were not associated with platelet 5-HT indices. Haplotypes at SLC6A4 and individual genotypes of polymorphisms at SLC6A4, HTR7, HTR2A, ITGB3, and TPH1 showed no significant association with whole blood 5-HT. Haplotype analysis of two polymorphisms in TPH1 revealed a nominally significant association with whole blood 5-HT (p=0.046). These initial studies of indices of the 5-HT system with several single-nucleotide polymorphisms at loci in this system generate hypotheses for testing in other samples. PMID- 17406649 TI - Bilateral hippocampal volume increase in patients with bipolar disorder and short term lithium treatment. AB - Most previous magnetic resonance imaging (MRI) studies of patients with bipolar disorder (BD) report similar hippocampus (HC) volumes across patients and controls, but because patients studied were heterogeneous with respect to course of illness variables and medication status, the conclusions of these studies remain equivocal. Lithium (Li) is the reference-standard drug for BD and its role as an important agent in neuroprotection and neurogenesis has been documented in human and in animal studies. We compared the volume of the HC, hippocampal head (Hh), and body/tail (Hbt) in three groups with no history of medication use before entry into this study: (a) a group of patients treated with Li for 1-8 weeks and then scanned; (b) a group comprised of patients who were unmedicated at the time of scan; and (c) a group of patients treated with either valproic acid or lamotrigine. Healthy age- and sex-matched comparison subjects were also scanned. HC volumes did not differ between the unmedicated and healthy comparison groups. There was a bilateral increase in volumes of HC and Hh in the Li-treated group compared to the unmedicated group, an effect that was apparent even over a brief treatment period. Our study provides further confirmation that Li can exert structural effects on the HC, which are detectable in vivo. The study emphasizes the need to control for even brief exposure to medication in volumetric studies of the HC. PMID- 17406650 TI - First human evidence of d-amphetamine induced displacement of a D2/3 agonist radioligand: A [11C]-(+)-PHNO positron emission tomography study. AB - Imaging the competition between D(2/3) radioligands and endogenous dopamine is so far the only way to measure dopamine release in the living human brain. The dopamine D(2) receptor exists in a high (D(2)(high)) and a low-affinity state for dopamine. Under physiological conditions, dopamine is expected to bind to D(2)(high) only. [(11)C]-(+)-4-propyl-9-hydroxynaphthoxazine ((+)-PHNO) is the first D(2/3) agonist radioligand for positron emission tomography (PET) imaging in humans. Since [(11)C]-(+)-PHNO is expected to bind preferentially to D(2)(high), it should be particularly vulnerable to competition with endogenous dopamine. Nine healthy subjects participated in two PET scans, one after administration of d-amphetamine and one after placebo. [(11)C]-(+)-PHNO PET test re-test variability was determined in 11 healthy subjects. Binding potentials (BPs) were calculated for caudate, putamen, ventral striatum, and globus pallidus. d-Amphetamine led to a significant decrease of [(11)C]-(+)-PHNO BPs in caudate (-13.2%), putamen (-20.8%), and ventral striatum (-24.9%), but not in globus pallidus (-6.5%). d-Amphetamine-induced displacement correlated with serum d-amphetamine levels in all regions but caudate. This is the first report on competition between endogenous dopamine and a D(2/3) agonist radioligand in humans. [(11)C]-(+)-PHNO PET might be a superior measure for release of endogenous dopamine than PET employing conventional D(2/3) antagonist radioligands. PMID- 17406651 TI - Maximizing the adequacy of medication treatment in controlled trials and clinical practice: STAR(*)D measurement-based care. AB - The success of well-developed protocols has been limited in real-world practice, where even effective strategies have not been sufficient to meet patient needs in routine clinical care owing to Axis I and III comorbidities. The Sequenced Treatment Alternatives to Relieve Depression (STAR(*)D) trial required that antidepressant medication treatment be optimal regarding dose and duration, yet accommodate flexibility to ensure safety given the wide range of comorbid general medical and psychiatric disorders allowed in the trial. The objective of this study was to develop a measurement-based care (MBC) approach and an automated feedback system to ensure adequate and safe antidepressant treatment delivery suitable for both clinical research and routine practice. Ratings of depressive symptom severity and side-effect frequency, intensity, and burden were obtained at each treatment visit using the MBC system that (1) guided medication dose adjustments and treatment duration, (2) documented clinician adherence to treatment recommendations, and (3) provided prompt feedback to clinicians to enhance appropriate treatment decisions. Physician adherence to protocol-specific treatment was monitored based on measured symptoms and side-effect burden, and dose and duration of antidepressant at each critical decision point during the acute phase treatment of major depression. Feedback was provided at the point of care by the clinical coordinators, assisted by Web-based reports following each treatment visit. On the basis of the first treatment step with citalopram, over 85% of treatment encounters had appropriate fidelity to recommendations. Most deviations from treatment recommendations occurred late in treatment and were often justifiable. MBC proved to be feasible and effective in busy primary and psychiatric settings. This approach signals a paradigm shift toward the use of measurement-based clinical decisions, both at the point of care and following each visit, to deliver optimal pharmacotherapy for depression. PMID- 17406652 TI - Minocycline attenuates neuronal cell death and improves cognitive impairment in Alzheimer's disease models. AB - Minocycline is a semi-synthetic tetracycline antibiotic that effectively crosses the blood-brain barrier. Minocycline has been reported to have significant neuroprotective effects in models of cerebral ischemia, traumatic brain injury, amyotrophic lateral sclerosis, and Huntington's and Parkinson's diseases. In this study, we demonstrate that minocycline has neuroprotective effects in in vitro and in vivo Alzheimer's disease models. Minocycline was found to attenuate the increases in the phosphorylation of double-stranded RNA-dependent serine/threonine protein kinase, eukaryotic translation initiation factor-2 alpha and caspase 12 activation induced by amyloid beta peptide1-42 treatment in NGF differentiated PC 12 cells. In addition, increases in the phosphorylation of eukaryotic translation initiation factor-2 alpha were attenuated by administration of minocycline in Tg2576 mice, which harbor mutated human APP695 gene including the Swedish double mutation and amyloid beta peptide(1-42)-infused rats. We found that minocycline administration attenuated deficits in learning and memory in amyloid beta peptide(1-42)-infused rats. Increased phosphorylated state of eukaryotic translation initiation factor-2 alpha is observed in Alzheimer's disease patients' brains and may result in impairment of cognitive functions in Alzheimer's disease patients by decreasing the efficacy of de novo protein synthesis required for synaptic plasticity. On the basis of these results, minocycline may prove to be a good candidate as an effective therapeutic agent for Alzheimer's disease. PMID- 17406653 TI - Endocannabinoid hedonic hotspot for sensory pleasure: anandamide in nucleus accumbens shell enhances 'liking' of a sweet reward. AB - Cannabinoid drugs such as Delta9-THC are euphoric and rewarding, and also stimulate food intake in humans and animals. Little is known about how naturally occurring endogenous brain cannabinoids mediate pleasure from food or other natural sensory rewards. The taste reactivity paradigm measures effects of brain manipulations on affective orofacial reactions to intraorally administered pleasant and unpleasant tastes. Here we tested if anandamide microinjection into medial nucleus accumbens shell enhances these affective reactions to sweet and bitter tastes in rats. Anandamide doubled the number of positive 'liking' reactions elicited by intraoral sucrose, without altering negative 'disliking' reactions to bitter quinine. Anandamide microinjections produced Fos plumes of approximately 0.02-1 mm3 volume. Plume-based maps, integrated with behavioral data, identified the medial shell of accumbens as the anatomical hotspot responsible for hedonic amplification. Anandamide produced especially intense hedonic enhancement in a roughly 1.6 mm3 'hedonic hotspot' in dorsal medial shell, where anandamide also stimulated eating behavior. These results demonstrate that endocannabinoid signals within medial accumbens shell specifically amplify the positive hedonic impact of a natural reward (though identification of the receptor specificity of this effect will require future studies). Identification of an endocannabinoid hotspot for sensory pleasure gives insight into brain mechanisms of natural reward, and may be relevant to understanding the neural effects of cannabinoid drugs of abuse and therapeutic agents. PMID- 17406654 TI - Evaluation of reference regions for (R)-[(11)C]PK11195 studies in Alzheimer's disease and mild cognitive impairment. AB - Inflammation in Alzheimer's disease (AD) may be assessed using (R)-[(11)C]PK11195 and positron emission tomography. Data can be analyzed using the simplified reference tissue model, provided a suitable reference region is available. This study evaluates various reference regions for analyzing (R)-[(11)C]PK11195 scans in patients with mild cognitive impairment (MCI) and probable AD. Healthy subjects (n=10, 30+/-10 years and n=10, 70+/-6 years) and patients with MCI (n=10, 74+/-6 years) and probable AD (n=9, 71+/-6 years) were included. Subjects underwent a dynamic three-dimensional (R)-[(11)C]PK11195 scan including arterial sampling. Gray matter, white matter, total cerebellum and cerebrum, and cluster analysis were evaluated as reference regions. Both plasma input binding potentials of these reference regions (BP(PLASMA)) and corresponding reference region input binding potentials of a target region (BP(SRTM)) were evaluated. Simulations were performed to assess cluster analysis performance at 5% to 15% coefficient of variation noise levels. Reasonable correlations for BP(PLASMA) (R(2)=0.52 to 0.94) and BP(SRTM) (R(2)=0.59 to 0.76) were observed between results using anatomic regions and cluster analysis. For cerebellum white matter, cerebrum white matter, and total cerebrum a considerable number of unrealistic BP(SRTM) values were observed. Cluster analysis did not extract a valid reference region in 10% of the scans. Simulations showed that potentially cluster analysis suffers from negative bias in BP(PLASMA). Most anatomic regions outperformed cluster analysis in terms of absence of both scan rejection and bias. Total cerebellum is the optimal reference region in this patient category. PMID- 17406655 TI - TNF alpha and Fas mediate tissue damage and functional outcome after traumatic brain injury in mice. AB - Tumor necrosis factor-alpha (TNFalpha) and Fas are induced after traumatic brain injury (TBI); however, their functional roles are incompletely understood. Using controlled cortical impact (CCI) and mice deficient in TNFalpha, Fas, or both (TNFalpha/Fas-/-), we hypothesized that TNFalpha and Fas receptor mediate secondary TBI in a redundant manner. Compared with wild type (WT), TNFalpha/Fas-/ mice had improved motor performance from 1 to 4 days (P<0.05), improved spatial memory acquisition at 8 to 14 days (P<0.05), and decreased brain lesion size at 2 and 6 weeks after CCI (P<0.05). Protection in TNFalpha/Fas-/- mice from histopathological and motor deficits was reversed by reconstitution with recombinant TNFalpha before CCI, and TNFalpha-/- mice administered anti-Fas ligand antibodies had improved spatial memory acquisition versus similarly treated WT mice (P<0.05). Tumor necrosis factor-alpha/Fas-/- mice had decreased the numbers of cortical cells with plasmalemma damage at 6 h (P<0.05 versus WT), and reduced matrix metalloproteinase-9 activity in injured brain at 48 and 72 h after CCI. In immature mice subjected to CCI, genetic inhibition of TNFalpha and Fas conferred beneficial effects on histopathology and spatial memory acquisition in adulthood (both P<0.05 versus WT), suggesting that the beneficial effects of TNFalpha/Fas inhibition may be permanent. The data suggest that redundant signaling pathways initiated by TNFalpha and Fas play pivotal roles in the pathogenesis of TBI, and that biochemical mechanisms downstream of TNFalpha/Fas may be novel therapeutic targets to limit neurological sequelae in children and adults with severe TBI. PMID- 17406656 TI - Androgenic/estrogenic balance in the male rat cerebral circulation: metabolic enzymes and sex steroid receptors. AB - Tissues from males can be regulated by a balance of androgenic and estrogenic effects because of local metabolism of testosterone and expression of relevant steroid hormone receptors. As a critical first step to understanding sex hormone influences in the cerebral circulation of males, we investigated the presence of enzymes that metabolize testosterone to active products and their respective receptors. We found that cerebral blood vessels from male rats express 5alpha reductase type 2 and aromatase, enzymes responsible for conversion of testosterone into dihydrotestosterone (DHT) and 17beta-estradiol, respectively. Protein levels of these enzymes, however, were not modulated by long-term in vivo hormone treatment. We also showed the presence of receptors for both androgens (AR) and estrogens (ER) from male cerebral vessels. Western blot analysis showed bands corresponding to the full-length AR (110 kDa) and ERalpha (66 kDa). Long term in vivo treatment of orchiectomized rats with testosterone or DHT, but not estrogen, increased AR levels in cerebral vessels. In contrast, ERalpha protein levels were increased after in vivo treatment with estrogen but not testosterone. Fluorescent immunostaining revealed ERalpha, AR, and 5alpha-reductase type 2 in both the endothelial and smooth muscle layers of cerebral arteries, whereas aromatase staining was solely localized to the endothelium. Thus, cerebral vessels from males are target tissues for both androgens and estrogen. Furthermore, local metabolism of testosterone might balance opposing androgenic and estrogenic influences on cerebrovascular as well as brain function in males. PMID- 17406657 TI - Reduced HO-1 protein expression is associated with more severe neurodegeneration after transient ischemia induced by cortical compression in diabetic Goto Kakizaki rats. AB - Pronounced hyperglycemia provoked by extradural compression (EC) of the sensorimotor cortex was recently described in the non-insulin dependent Goto Kakizaki (GK) diabetic rat. Compared with control Wistar rats, GK rats exhibited more extensive brain damage after cortical ischemia at 48 h of reperfusion (Moreira et al, 2007). We hypothesized that the enhanced brain injury in GK rats could be caused by differential regulation of the heme degrading enzyme heme oxygenase (HO)-1, known to interact with the expression of other target genes implicated in antioxidant defense, inflammation and neurodegeneration, such as superoxide dismutase (SOD)-1, -2, inducible nitric oxide synthase (iNOS), and tumor necrosis factor-alpha (TNFalpha). At 48 h after ischemia, relative mRNA expression of such target genes was compared between ipsilateral (compressed) and contralateral (uncompressed) hemispheres of GK rats, along with baseline comparison of sham, uncompressed GK and Wistar rats. Immunohistochemistry was performed to detect cellular and regional localization of HO-1 at this time point. Baseline expression of HO-1, iNOS, and TNFalpha mRNA was increased in the cortex of sham GK rats. GK rats showed pronounced hyperglycemia during EC and transient attenuation of regional cerebral blood flow recovery. At 48 h after reperfusion, HO-1 mRNA expression was 7- to 8-fold higher in the ischemic cortex of both strains, being the most upregulated gene under study. Heme oxygenase-1 protein expression was significantly reduced in diabetic rats and was found in perilesional astrocytes and rare microglial cells, in both strains. The reduced HO-1 protein expression in GK rats at 48 h after reperfusion combined with more extensive neurodegeneration induced by EC, provides further in vivo evidence for a neuroprotective role of HO after brain ischemia. PMID- 17406658 TI - Cerebrovascular effects of hemodialysis in chronic kidney disease. AB - Patients with end-stage renal disease (ESRD) undergoing hemodialysis are known to suffer cognitive deficits and stroke of unknown etiology. It has been suspected that the treatment itself may contribute to the syndrome by unknown mechanisms, which we investigated in this study. End-stage renal disease patients on hemodialysis (n=19) or peritoneal dialysis (PD, n=5) were compared with 14 healthy controls. Subjects participated in magnetic resonance imaging (MRI) measurements of cerebral atrophy, cerebral blood flow (CBF) arterial spin labeled MRI (ASL-MRI), quantitative Doppler blood flow through the internal carotid artery, and cerebral oxymetry. The Doppler and oxymetry procedures were also performed at the beginning and end of a single hemodialysis session. End-stage renal disease patients on hemodialysis showed significant cerebral atrophy, associated with longer hemodialysis duration and cognitive deficits, including focal bilateral lesions in the caudate nucleus and midbrain. Cerebral oxygenation was extremely low before dialysis (rSO(2) 41+/-13, compared with 70+/-2 in controls, P<0.02) and improved only slightly after dialysis. Carotid blood flow was also very low at the start of dialysis (115+/-28 mL/sec, versus 193+/-56 in controls, P<0.005) but normalized at the end of the session (181 mL/sec). The PD patients showed intermediate values, between the hemodialysis and controls. Notably, duration of hemodialysis treatment predicted global gray-matter volume (r=-0.74), change of blood flow during dialysis (r=-0.65), and baseline rSO(2) (r=-0.65). The findings suggest that ESRD patients on hemodialysis suffer low CBF during the interdialytic cycle. Coupled with low cerebral oxygenation levels and atherosclerosis, this may contribute significantly to the etiology of the observed cerebral atrophy, cognitive deficits, and high stroke prevalence. PMID- 17406659 TI - Dynamic forcing of end-tidal carbon dioxide and oxygen applied to functional magnetic resonance imaging. AB - Investigations into the blood oxygenation level-dependent (BOLD) functional MRI signal have used respiratory challenges with the aim of probing cerebrovascular physiology. Such challenges have altered the inspired partial pressures of either carbon dioxide or oxygen, typically to a fixed and constant level (fixed inspired challenge (FIC)). The resulting end-tidal gas partial pressures then depend on the subject's metabolism and ventilatory responses. In contrast, dynamic end tidal forcing (DEF) rapidly and independently sets end-tidal oxygen and carbon dioxide to desired levels by altering the inspired gas partial pressures on a breath-by-breath basis using computer-controlled feedback. This study implements DEF in the MRI environment to map BOLD signal reactivity to CO(2). We performed BOLD (T2(*)) contrast FMRI in four healthy male volunteers, while using DEF to provide a cyclic normocapnic-hypercapnic challenge, with each cycle lasting 4 mins (PET(CO(2)) mean+/-s.d., from 40.9+/-1.8 to 46.4+/-1.6 mm Hg). This was compared with a traditional fixed-inspired (FI(CO(2))=5%) hypercapnic challenge (PET(CO(2)) mean+/-s.d., from 38.2+/-2.1 to 45.6+/-1.4 mm Hg). Dynamic end-tidal forcing achieved the desired target PET(CO(2)) for each subject while maintaining PET(O(2)) constant. As a result of CO(2)-induced increases in ventilation, the FIC showed a greater cyclic fluctuation in PET(O(2)). These were associated with spatially widespread fluctuations in BOLD signal that were eliminated largely by the control of PET(O(2)) during DEF. The DEF system can provide flexible, convenient, and physiologically well-controlled respiratory challenges in the MRI environment for mapping dynamic responses of the cerebrovasculature. PMID- 17406660 TI - Limitations on adaptation: evolutionary response to climatic change? PMID- 17406661 TI - Human expression patterns: genetic differences between populations. PMID- 17406663 TI - Towards woven logic from organic electronic fibres. AB - The use of organic polymers for electronic functions is mainly motivated by the low-end applications, where low cost rather than advanced performance is a driving force. Materials and processing methods must allow for cheap production. Printing of electronics using inkjets or classical printing methods has considerable potential to deliver this. Another technology that has been around for millennia is weaving using fibres. Integration of electronic functions within fabrics, with production methods fully compatible with textiles, is therefore of current interest, to enhance performance and extend functions of textiles. Standard polymer field-effect transistors require well defined insulator thickness and high voltage, so they have limited suitability for electronic textiles. Here we report a novel approach through the construction of wire electrochemical transistor (WECT) devices, and show that textile monofilaments with 10-100 mum diameters can be coated with continuous thin films of the conducting polythiophene poly(3,4-ethylenedioxythiophene), and used to create micro-scale WECTs on single fibres. We also demonstrate inverters and multiplexers for digital logic. This opens an avenue for three-dimensional polymer micro-electronics, where large-scale circuits can be designed and integrated directly into the three-dimensional structure of woven fibres. PMID- 17406662 TI - Quantitative trait loci linked to thalamus and cortex gray matter volumes in BXD recombinant inbred mice. AB - To investigate whether there are separate or shared genetic influences on the development of the thalamus and cerebral cortex, we identified quantitative trait loci (QTLs) for relevant structural volumes in BXD recombinant inbred (RI) strains of mice. In 34 BXD RI strains and two parental strains (C57BL/6J and DBA/2J), we measured the volumes of the entire thalamus and cortex gray matter using point counting and Cavalieri's rule. Heritability was calculated using analysis of variance (ANOVA), and QTL analysis was carried out using WebQTL (http://www.genenetwork.org). The heritability of thalamus volume was 36%, and three suggestive QTLs for thalamus volume were identified on chromosomes 10, 11 and 16. The heritability of cortical gray matter was 43%, and four suggestive QTLs for cortex gray matter volume were identified on chromosomes 2, 8, 16 and 19. The genetic correlation between thalamus and cortex gray matter volumes was 0.64. Also, a single QTL on chromosome 16 (D16Mit100) was identified for thalamus volume, cortex gray matter volume and Morris water maze search-time preference (r=0.71). These results suggest that there are separate and shared genetic influences on the development of the thalamus and cerebral cortex. PMID- 17406664 TI - Role of ubiquitination in IGF-1 receptor signaling and degradation. AB - BACKGROUND: The insulin-like growth factor 1 receptor (IGF-1R) plays numerous crucial roles in cancer biology. The majority of knowledge on IGF-1R signaling is concerned with its role in the activation of the canonical phosphatidyl inositol 3 kinase (PI3K)/Akt and MAPK/ERK pathways. However, the role of IGF-1R ubiquitination in modulating IGF-1R function is an area of current research. In light of this we sought to determine the relationship between IGF-1R phosphorylation, ubiquitination, and modulation of growth signals. METHODOLOGY: Wild type and mutant constructs of IGF-1R were transfected into IGF-1R null fibroblasts. IGF-1R autophosphorylation and ubiquitination were determined by immunoprecipitation and western blotting. IGF-1R degradation and stability was determined by cyclohexamide-chase assay in combination with lysosome and proteasome inhibitors. PRINCIPAL FINDINGS: IGF-1R autophosphorylation was found to be an absolute requirement for receptor ubiquitination. Deletion of C-terminal domain had minimal effect on IGF-1 induced receptor autophosphorylation, however, ubiquitination and ERK activation were completely abolished. Cells expressing kinase impaired IGF-1R, exhibited both receptor ubiquitination and ERK phosphorylation, however failed to activate Akt. While IGF-1R mutants with impaired PI3K/Akt signaling were degraded mainly by the proteasomes, the C terminal truncated one was exclusively degraded through the lysosomal pathway. CONCLUSIONS: Our data suggest important roles of ubiquitination in mediating IGF 1R signaling and degradation. Ubiquitination of IGF-1R requires receptor tyrosine kinase activity, but is not involved in Akt activation. In addition we show that the C-terminal domain of IGF-1R is a necessary requisite for ubiquitination and ERK phosphorylation as well as for proteasomal degradation of the receptor. PMID- 17406665 TI - Daytime naps, motor memory consolidation and regionally specific sleep spindles. AB - BACKGROUND: Increasing evidence demonstrates that motor-skill memories improve across a night of sleep, and that non-rapid eye movement (NREM) sleep commonly plays a role in orchestrating these consolidation enhancements. Here we show the benefit of a daytime nap on motor memory consolidation and its relationship not simply with global sleep-stage measures, but unique characteristics of sleep spindles at regionally specific locations; mapping to the corresponding memory representation. METHODOLOGY/PRINCIPAL FINDINGS: Two groups of subjects trained on a motor-skill task using their left hand - a paradigm known to result in overnight plastic changes in the contralateral, right motor cortex. Both groups trained in the morning and were tested 8 hr later, with one group obtaining a 60 90 minute intervening midday nap, while the other group remained awake. At testing, subjects that did not nap showed no significant performance improvement, yet those that did nap expressed a highly significant consolidation enhancement. Within the nap group, the amount of offline improvement showed a significant correlation with the global measure of stage-2 NREM sleep. However, topographical sleep spindle analysis revealed more precise correlations. Specifically, when spindle activity at the central electrode of the non-learning hemisphere (left) was subtracted from that in the learning hemisphere (right), representing the homeostatic difference following learning, strong positive relationships with offline memory improvement emerged-correlations that were not evident for either hemisphere alone. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that motor memories are dynamically facilitated across daytime naps, enhancements that are uniquely associated with electrophysiological events expressed at local, anatomically discrete locations of the brain. PMID- 17406666 TI - Mad3 KEN boxes mediate both Cdc20 and Mad3 turnover, and are critical for the spindle checkpoint. AB - Mitotic progression is controlled by proteolytic destruction of securin and cyclin. The mitotic E3 ubiquitin ligase, known as the anaphase promoting complex or cyclosome (APC/C), in partnership with its activators Cdc20p and Cdh1p, targets these proteins for degradation. In the presence of defective kinetochore microtubule interactions, APC/C(Cdc20) is inhibited by the spindle checkpoint, thereby delaying anaphase onset and providing more time for spindle assembly. Cdc20p interacts directly with Mad2p, and its levels are subject to careful regulation, but the precise mode(s) of APC/C( Cdc20) inhibition remain unclear. The mitotic checkpoint complex (MCC, consisting of Mad3p, Mad2p, Bub3p and Cdc20p in budding yeast) is a potent APC/C inhibitor. Here we focus on Mad3p and how it acts, in concert with Mad2p, to efficiently inhibit Cdc20p. We identify and analyse the function of two motifs in Mad3p, KEN30 and KEN296, which are conserved from yeast Mad3p to human BubR1. These KEN amino acid sequences resemble 'degron' signals that confer interaction with APC/C activators and target proteins for degradation. We show that both Mad3p KEN boxes are necessary for spindle checkpoint function. Mutation of KEN30 abolished MCC formation and stabilised Cdc20p in mitosis. In addition, mutation of Mad3-KEN30, APC/C subunits, or Cdh1p, stabilised Mad3p in G1, indicating that the N-terminal KEN box could be a Mad3p degron. To determine the significance of Mad3p turnover, we analysed the consequences of MAD3 overexpression and found that four-fold overproduction of Mad3p led to chromosome bi-orientation defects and significant chromosome loss during recovery from anti-microtubule drug induced checkpoint arrest. In conclusion, Mad3p KEN30 mediates interactions that regulate the proteolytic turnover of Cdc20p and Mad3p, and the levels of both of these proteins are critical for spindle checkpoint signaling and high fidelity chromosome segregation. PMID- 17406667 TI - Onset rivalry: brief presentation isolates an early independent phase of perceptual competition. AB - When the left and right eyes are simultaneously presented with different images, observers typically report exclusive awareness of only one image. This phenomenon is termed binocular rivalry, reflecting the fact that the dominant image alternates every few seconds in a cycle of perceptual competition that continues indefinitely. Despite the apparent continuity in perceptual switching, we now demonstrate that the initial "onset" period is fundamentally different to all subsequent rivalry epochs. Using brief intermittent presentations, rivalry dominance shows strong biases such that the same target is perceived with each successive stimulus onset. These biases remain consistent within any given location, but vary across the visual field in a distribution that is stable over multiple weeks but highly idiosyncratic across observers. If the presentation exceeds approximately 1sec at any location, however, the very different and much more balanced alternations of sustained binocular rivalry become apparent. These powerful onset biases are observed with brief intermittent presentations at a single location or with continual smooth motion of the targets. Periods of adaptation to one of the rivaling targets induced local switches in dominance to the non-adapted target. However, these effects were generally limited to the spatial site of adaptation and had less influence over each subsequent cycle of the target. We conclude that onset rivalry is independent of sustained rivalry and cannot be explained by local regions of monocular dominance or memory of past perceptual history, but rather reflects low-level, spatially localized factors that are stable over periods of weeks. These findings suggest that brief presentation paradigms are inappropriate for their current use in studies of the mechanisms underlying sustained rivalry. However, brief presentations are ideal for investigating early stages of perceptual competition. PMID- 17406668 TI - Guillain-Barre syndrome and preceding infection with campylobacter, influenza and Epstein-Barr virus in the general practice research database. AB - BACKGROUND: A number of infectious agents have previously been suggested as risk factors for the development of Guillain-Barre syndrome (GBS), but robust epidemiologic evidence for these associations is lacking. METHODS AND FINDINGS: We conducted a nested case-control study using data from the United Kingdom General Practice Research Database between 1991 and 2001. Controls were matched to cases on general practice clinic, sex, year of birth and date of outcome diagnosis in their matched case. We found positive associations between GBS and infection with Campylobacter, Epstein-Barr virus and influenza-like illness in the previous two months, as well as evidence of a protective effect of influenza vaccination. After correction for under-ascertainment of Campylobacter infection, the excess risk of GBS following Campylobacter enteritis was 60-fold and 20% of GBS cases were attributable to this pathogen. CONCLUSIONS: Our findings indicate a far greater excess risk of GBS among Campylobacter enteritis patients than previously reported by retrospective serological studies. In addition, they confirm previously suggested associations between infection due to Epstein-Barr virus infection and influenza-like illness and GBS. Finally, we report evidence of a protective effect of influenza vaccination on GBS risk, which may be mediated through protection against influenza disease, or result from a lower likelihood of vaccination among those with recent infection. Cohort studies of GBS incidence in this population would help to clarify the burden of GBS due to influenza, and any potential protective effect of influenza vaccination. PMID- 17406669 TI - Compartmental architecture and dynamics of hematopoiesis. AB - BACKGROUND: Blood cell formation is maintained by the replication of hematopoietic stem cells (HSC) that continuously feed downstream "compartments" where amplification and differentiation of cells occurs, giving rise to all blood lineages. Whereas HSC replicate slowly, committed cells replicate faster as they become more differentiated. METHODOLOGY/SIGNIFICANT FINDING: We propose a multi compartment model of hematopoiesis, designed on the principle of cell flow conservation under stationary conditions. Cells lost from one compartment due to differentiation are replaced by cells from the upstream compartment. We assume that there is a constant relationship between cell input and output in each compartment and fix the single parameter of the model using data available for granulocyte maturation. We predict that approximately 31 mitotic events separate the HSC from the mature cells observed in the circulation. Besides estimating the number of compartments, our model allows us to estimate the size of each compartment, the rate of cell replication within each compartment, the mean time a given cell type contributes to hematopoiesis, the amplification rate in each compartment, as well as the mean time separating stem-cell replication and mature blood-cell formation. CONCLUSIONS: Despite its simplicity, the model agrees with the limited in vivo data available and can make testable predictions. In particular, our prediction of the average lifetime of a PIG-A mutated clone agrees closely with the experimental results available for the PIG-A gene mutation in healthy adults. The present elucidation of the compartment structure and dynamics of hematopoiesis may prove insightful in further understanding a variety of hematopoietic disorders. PMID- 17406670 TI - Environmental enrichment effects on development of retinal ganglion cell dendritic stratification require retinal BDNF. AB - A well-known developmental event of retinal maturation is the progressive segregation of retinal ganglion cell (RGC) dendrites into a and b sublaminae of the inner plexiform layer (IPL), a morphological rearrangement crucial for the emergence of the ON and OFF pathways. The factors regulating this process are not known, although electrical activity has been demonstrated to play a role. Here we report that Environmental Enrichment (EE) accelerates the developmental segregation of RGC dendrites and prevents the effects exerted on it by dark rearing (DR). Development of RGC stratification was analyzed in a line of transgenic mice expressing plasma-membrane marker green fluorescent protein (GFP) under the control of Thy-1 promoter; we visualized the a and b sublaminae of the IPL by using an antibody selectively directed against a specific marker of cholinergic neurons. EE precociously increases Brain Derived Neurotrophic Factor (BDNF) in the retina, in parallel with the precocious segregation of RGC dendrites; in addition, EE counteracts retinal BDNF reduction in DR retinas and promotes a normal segregation of RGC dendrites. Blocking retinal BDNF by means of antisense oligos blocks EE effects on the maturation of RGC dendritic stratification. Thus, EE affects the development of RGC dendritic segregation and retinal BDNF is required for this effect to take place, suggesting that BDNF could play an important role in the emergence of the ON and OFF pathways. PMID- 17406671 TI - Glycosaminoglycan interactions in murine gammaherpesvirus-68 infection. AB - Glycosaminoglycans (GAGs) commonly participate in herpesvirus entry. They are thought to provide a reversible attachment to cells that promotes subsequent receptor binding. Murine gamma-herpesvirus-68 (MHV-68) infection of fibroblasts and epithelial cells is highly GAG-dependent. This is a function of the viral gp150, in that gp150-deficient mutants are much less GAG-dependent than wild type. Here we show that the major MHV-68 GAG-binding protein is not gp150 but gp70, a product of ORF4. Surprisingly, ORF4-deficient MHV-68 showed normal cell binding and was more sensitive than wild-type to inhibition by soluble heparin rather than less. Thus, the most obvious viral GAG interaction made little direct contribution to infection. Indeed, a large fraction of the virion gp70 had its GAG-binding domain removed by post-translational cleavage. ORF4 may therefore act mainly to absorb soluble GAGs and prevent them from engaging gp150 prematurely. In contrast to gp70, gp150 bound poorly to GAGs, implying that it provides little in the way of adhesion. We hypothesize that it acts instead as a GAG-sensitive switch that selectively activates MHV-68 entry at cell surfaces. PMID- 17406672 TI - Why are male social relationships complex in the Doubtful Sound bottlenose dolphin population? AB - BACKGROUND: Access to oestrus females tends to be the main driver of male sociality. This factor can lead to complex behavioural interactions between males and groups of males. Male bottlenose dolphins may form alliances to consort females and to compete with other males. In some populations these alliances may form temporary coalitions when competing for females. I examined the role of dyadic and group interactions in the association patterns of male bottlenose dolphins in Doubtful Sound, New Zealand. There is no apparent mating competition in this population and no consortship has been observed, yet agonistic interactions between males occur regularly. METHODOLOGY/PRINCIPAL FINDINGS: By comparing the network of male interactions in several social dimensions (affiliative, agonistic, and associative) I show that while agonistic interactions relate to dyadic association patterns, affiliative interactions seem to relate to group association patterns. Some evidence suggests that groups of males also formed temporary coalitions during agonistic interactions. While different groups of males had similar relationships with non-oestrus females, the time they spent with oestrus females and mothers of newborns differed greatly. CONCLUSIONS/SIGNIFICANCE: After considering several hypotheses, I propose that the evolution of these complex relationships was driven by sexual competition probably to out-compete other males for female choice. PMID- 17406673 TI - The transmissibility of highly pathogenic avian influenza in commercial poultry in industrialised countries. AB - BACKGROUND: With the increased occurrence of outbreaks of H5N1 worldwide there is concern that the virus could enter commercial poultry farms with severe economic consequences. METHODOLOGY/PRINCIPAL FINDINGS: We analyse data from four recent outbreaks of highly pathogenic avian influenza (HPAI) in commercial poultry to estimate the farm-to-farm reproductive number for HPAI. The reproductive number is a key measure of the transmissibility of HPAI at the farm level because it can be used to evaluate the effectiveness of the control measures. In these outbreaks the mean farm-to-farm reproductive number prior to controls ranged from 1.1 to 2.4, with the maximum farm-based reproductive number in the range 2.2 to 3.2. Enhanced bio-security, movement restrictions and prompt isolation of the infected farms in all four outbreaks substantially reduced the reproductive number, but it remained close to the threshold value 1 necessary to ensure the disease will be eradicated. CONCLUSIONS/SIGNIFICANCE: Our results show that depending on the particular situation in which an outbreak of avian influenza occurs, current controls might not be enough to eradicate the disease, and therefore a close monitoring of the outbreak is required. The method we used for estimating the reproductive number is straightforward to implement and can be used in real-time. It therefore can be a useful tool to inform policy decisions. PMID- 17406674 TI - Metabolic engineering of potato carotenoid content through tuber-specific overexpression of a bacterial mini-pathway. AB - BACKGROUND: Since the creation of "Golden Rice", biofortification of plant derived foods is a promising strategy for the alleviation of nutritional deficiencies. Potato is the most important staple food for mankind after the cereals rice, wheat and maize, and is extremely poor in provitamin A carotenoids. METHODOLOGY: We transformed potato with a mini-pathway of bacterial origin, driving the synthesis of beta-carotene (Provitamin A) from geranylgeranyl diphosphate. Three genes, encoding phytoene synthase (CrtB), phytoene desaturase (CrtI) and lycopene beta-cyclase (CrtY) from Erwinia, under tuber-specific or constitutive promoter control, were used. 86 independent transgenic lines, containing six different promoter/gene combinations, were produced and analyzed. Extensive regulatory effects on the expression of endogenous genes for carotenoid biosynthesis are observed in transgenic lines. Constitutive expression of the CrtY and/or CrtI genes interferes with the establishment of transgenosis and with the accumulation of leaf carotenoids. Expression of all three genes, under tuber specific promoter control, results in tubers with a deep yellow ("golden") phenotype without any adverse leaf phenotypes. In these tubers, carotenoids increase approx. 20-fold, to 114 mcg/g dry weight and beta-carotene 3600-fold, to 47 mcg/g dry weight. CONCLUSIONS: This is the highest carotenoid and beta carotene content reported for biofortified potato as well as for any of the four major staple foods (the next best event being "Golden Rice 2", with 31 mcg/g dry weight beta-carotene). Assuming a beta-carotene to retinol conversion of 6ratio1, this is sufficient to provide 50% of the Recommended Daily Allowance of Vitamin A with 250 gms (fresh weight) of "golden" potatoes. PMID- 17406675 TI - Affiliative behavior, ultrasonic communication and social reward are influenced by genetic variation in adolescent mice. AB - Social approach is crucial for establishing relationships among individuals. In rodents, social approach has been studied primarily within the context of behavioral phenomena related to sexual reproduction, such as mating, territory defense and parental care. However, many forms of social interaction occur before the onset of reproductive maturity, which suggests that some processes underlying social approach among juvenile animals are probably distinct from those in adults. We conducted a longitudinal study of social investigation (SI) in mice from two inbred strains to assess the extent to which genetic factors influence the motivation for young mice to approach one another. Early-adolescent C57BL/6J (B6) mice, tested 4-6 days after weaning, investigated former cage mates to a greater degree than BALB/cJ (BALB) mice, irrespective of the sex composition within an interacting pair. This strain difference was not due to variation in maternal care, the phenotypic characteristics of stimulus mice or sensitivity to the length of isolation prior to testing, nor was it attributable to a general difference in appetitive motivation. Ultrasonic vocalization (USV) production was positively correlated with the SI responses of mice from both strains. Interestingly, several USV characteristics segregated with the genetic background of young mice, including a higher average frequency and shorter duration for the USVs emitted by B6 mice. An assessment of conditioned place preference responses indicated that there was a strain-dependent difference in the rewarding nature of social contact. As adolescent mice aged, SI responses gradually became less sensitive to genetic background and more responsive to the particular sex of individuals within an interacting pair. We have thus identified a specific, genetic influence on the motivation of early-adolescent mice to approach one another. Consistent with classical theories of motivation, which propose a functional relationship between behavioral approach and reward, our findings indicate that reward is a proximal mechanism through which genetic factors affect social motivation during early adolescence. PMID- 17406676 TI - Genome sequencing shows that European isolates of Francisella tularensis subspecies tularensis are almost identical to US laboratory strain Schu S4. AB - BACKGROUND: Francisella tularensis causes tularaemia, a life-threatening zoonosis, and has potential as a biowarfare agent. F. tularensis subsp. tularensis, which causes the most severe form of tularaemia, is usually confined to North America. However, a handful of isolates from this subspecies was obtained in the 1980s from ticks and mites from Slovakia and Austria. Our aim was to uncover the origins of these enigmatic European isolates. METHODOLOGY/PRINCIPAL FINDINGS: We determined the complete genome sequence of FSC198, a European isolate of F. tularensis subsp. tularensis, by whole-genome shotgun sequencing and compared it to that of the North American laboratory strain Schu S4. Apparent differences between the two genomes were resolved by re sequencing discrepant loci in both strains. We found that the genome of FSC198 is almost identical to that of Schu S4, with only eight SNPs and three VNTR differences between the two sequences. Sequencing of these loci in two other European isolates of F. tularensis subsp. tularensis confirmed that all three European isolates are also closely related to, but distinct from Schu S4. CONCLUSIONS/SIGNIFICANCE: The data presented here suggest that the Schu S4 laboratory strain is the most likely source of the European isolates of F. tularensis subsp. tularensis and indicate that anthropogenic activities, such as movement of strains or animal vectors, account for the presence of these isolates in Europe. Given the highly pathogenic nature of this subspecies, the possibility that it has become established wild in the heartland of Europe carries significant public health implications. PMID- 17406677 TI - A study of T cell tolerance to the tumor-associated antigen MDM2: cytokines can restore antigen responsiveness, but not high avidity T cell function. AB - BACKGROUND: Most tumor-associated antigens (TAA) currently used for immunotherapy of cancer are also expressed in normal tissues, which may induce tolerance and impair T cell-mediated immunity. However, there is limited information about how physiological expression in normal tissues alters the function of TAA-specific T cells. METHODOLOGY/PRINCIPAL FINDINGS: We used a T cell receptor transgenic model to study how MDM2 expression in normal tissues affects the function of T cells specific for this TAA that is found at high levels in many different types of tumors. We found that some MDM2-specific T cells escaped thymic deletion and persisted in the peripheral T cell pool. When stimulated with antigen, these T cells readily initiated cell division but failed to proliferate and expand, which was associated with a high rate of apoptosis. Both IL-2 and IL-15 efficiently rescued T cell survival and antigen-specific T cell proliferation, while IL-7 and IL-21 were ineffective. Antigen-stimulated T cells showed impaired expression of the effector molecules CD43, granzyme-B and IFN-gamma, a defect that was completely restored when T cells were stimulated in the presence of IL-2. In contrast, IL-15 and IL-21 only restored the expression of CD43 and granzyme-B, but not IFN-gamma production. Finally, peptide titration experiments with IL-2 rescued T cells indicated that they were of lower avidity than non-tolerant control T cells expressing the same TCR. CONCLUSIONS/SIGNIFICANCE: These data indicate that cytokines can rescue the antigen-specific proliferation and effector function of MDM2-specific T cells, although this does not lead to the recovery of high avidity T cell function. This study sheds light on possible limitations of immunotherapy approaches that target widely expressed TAA, such as MDM2. PMID- 17406678 TI - Free-flight odor tracking in Drosophila is consistent with an optimal intermittent scale-free search. AB - During their trajectories in still air, fruit flies (Drosophila melanogaster) explore their landscape using a series of straight flight paths punctuated by rapid 90 degrees body-saccades [1]. Some saccades are triggered by visual expansion associated with collision avoidance. Yet many saccades are not triggered by visual cues, but rather appear spontaneously. Our analysis reveals that the control of these visually independent saccades and the flight intervals between them constitute an optimal scale-free active searching strategy. Two characteristics of mathematical optimality that are apparent during free-flight in Drosophila are inter-saccade interval lengths distributed according to an inverse square law, which does not vary across landscape scale, and 90 degrees saccade angles, which increase the likelihood that territory will be revisited and thereby reduce the likelihood that near-by targets will be missed. We also show that searching is intermittent, such that active searching phases randomly alternate with relocation phases. Behaviorally, this intermittency is reflected in frequently occurring short, slow speed inter-saccade intervals randomly alternating with rarer, longer, faster inter-saccade intervals. Searching patterns that scale similarly across orders of magnitude of length (i.e., scale free) have been revealed in animals as diverse as microzooplankton, bumblebees, albatrosses, and spider monkeys, but these do not appear to be optimised with respect to turning angle, whereas Drosophila free-flight search does. Also, intermittent searching patterns, such as those reported here for Drosophila, have been observed in foragers such as planktivorous fish and ground foraging birds. Our results with freely flying Drosophila may constitute the first reported example of searching behaviour that is both scale-free and intermittent. PMID- 17406679 TI - Multilevel deconstruction of the In vivo behavior of looped DNA-protein complexes. AB - Protein-DNA complexes with loops play a fundamental role in a wide variety of cellular processes, ranging from the regulation of DNA transcription to telomere maintenance. As ubiquitous as they are, their precise in vivo properties and their integration into the cellular function still remain largely unexplored. Here, we present a multilevel approach that efficiently connects in both directions molecular properties with cell physiology and use it to characterize the molecular properties of the looped DNA-lac repressor complex while functioning in vivo. The properties we uncover include the presence of two representative conformations of the complex, the stabilization of one conformation by DNA architectural proteins, and precise values of the underlying twisting elastic constants and bending free energies. Incorporation of all this molecular information into gene-regulation models reveals an unprecedented versatility of looped DNA-protein complexes at shaping the properties of gene expression. PMID- 17406680 TI - Extensive polycistronism and antisense transcription in the mammalian Hox clusters. AB - The Hox clusters play a crucial role in body patterning during animal development. They encode both Hox transcription factor and micro-RNA genes that are activated in a precise temporal and spatial sequence that follows their chromosomal order. These remarkable collinear properties confer functional unit status for Hox clusters. We developed the TranscriptView platform to establish high resolution transcriptional profiling and report here that transcription in the Hox clusters is far more complex than previously described in both human and mouse. Unannotated transcripts can represent up to 60% of the total transcriptional output of a cluster. In particular, we identified 14 non-coding Transcriptional Units antisense to Hox genes, 10 of which (70%) have a detectable mouse homolog. Most of these Transcriptional Units in both human and mouse present conserved sizeable sequences (>40 bp) overlapping Hox transcripts, suggesting that these Hox antisense transcripts are functional. Hox clusters also display at least seven polycistronic clusters, i.e., different genes being co transcribed on long isoforms (up to 30 kb). This work provides a reevaluated framework for understanding Hox gene function and dys-function. Such extensive transcriptions may provide a structural explanation for Hox clustering. PMID- 17406681 TI - Novelties in hybrid zones: crossroads between population genomic and ecological approaches. AB - BACKGROUND: Interspecific hybridization is widespread, occurring in a taxonomically diverse array of species. The Cyprinidae family, which displays more than 30% hybridization, is a good candidate for studies of processes underlying isolation and speciation, such as genetic exchange between previously isolated lineages. This is particularly relevant in the case of recent hybridization between an invasive species, Chondrostoma nasus nasus (from Eastern Europe), and C. toxostoma toxostoma (a threatened species endemic to southern France), in which bidirectional introgressive hybridization has been demonstrated. METHODOLOGY/PRINCIPAL FINDINGS: We studied 128 specimens from reference populations and 1495 hybrid zone specimens (two years of sampling and four stations), using five molecular markers (one mitochondrial gene, four nuclear introns), morphology (meristic and plastic characters) and life history traits (weight, size, coefficient of condition, sex, age, shoaling). We identified 65 hybrid combinations and visualized spatial and temporal changes in composition. The direction of mitochondrial introgression was density-dependent in favor of the rarer species and we demonstrate that the sexual selection hypothesis is a preponderant explanation in the asymmetry of introgression. Despite genomic evolution in the hybrid zone, convergence was observed for body shape and coefficient of condition, indicating changes in foraging behavior with respect to reference populations, reflecting strong environmental pressure. CONCLUSIONS/SIGNIFICANCE: The complex rules of hybrid zone dynamics are established very early in the contact zone. We propose "inheritance from the rare species" as a new evolutionary hypothesis for animal models. The endemic species was not assimilated by the invasive species. Survival rates for this species were highest in the middle of the river (the warmest part) due to a trade-off between food availability and fecundity. The environment-independent hybrid combination may result from nuclear-mitochondrial interactions involving the Tpi1b gene or a gene linked to this gene (Chromosome 16). This genomic region is also responsible for shoaling behavior in Danio rerio and is a promising zone for studies of changes in population dynamics and advances in integrated studies of hybrid zones. PMID- 17406682 TI - Modular mass spectrometric tool for analysis of composition and phosphorylation of protein complexes. AB - The combination of high accuracy, sensitivity and speed of single and multiple stage mass spectrometric analyses enables the collection of comprehensive sets of data containing detailed information about complex biological samples. To achieve these properties, we combined two high-performance matrix-assisted laser desorption ionization mass analyzers in one modular mass spectrometric tool, and applied this tool for dissecting the composition and post-translational modifications of protein complexes. As an example of this approach, we here present studies of the Saccharomyces cerevisiae anaphase-promoting complexes (APC) and elucidation of phosphorylation sites on its components. In general, the modular concept we describe could be useful for assembling mass spectrometers operating with both matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) ion sources into powerful mass spectrometric tools for the comprehensive analysis of complex biological samples. PMID- 17406684 TI - Design of chiral organocatalysts for practical asymmetric synthesis of amino acid derivatives. AB - A series of structurally rigid, chiral quaternary ammonium salts and several chiral sec-amine catalysts derived from commercially available (R)- or (S) binaphthol have been designed as new C(2)-symmetric chiral phase-transfer catalysts and chiral bifunctional amino-catalysts. These chiral organocatalysts have been successfully applied to the highly practical asymmetric synthesis of various amino acid derivatives. PMID- 17406683 TI - Evolutionary diversification of SPANX-N sperm protein gene structure and expression. AB - The sperm protein associated with nucleus in the X chromosome (SPANX) genes cluster at Xq27 in two subfamilies, SPANX-A/D and SPANX-N. SPANX-A/D is specific for hominoids and is fairly well characterized. The SPANX-N gave rise to SPANX A/D in the hominoid lineage approximately 7 MYA. Given the proposed role of SPANX genes in spermatogenesis, we have extended studies to SPANX-N gene evolution, variation, regulation of expression, and intra-sperm localization. By immunofluorescence analysis, SPANX-N proteins are localized in post-meiotic spermatids exclusively, like SPANX-A/D. But in contrast to SPANX-A/D, SPANX-N are found in all ejaculated spermatozoa rather than only in a subpopulation, are localized in the acrosome rather than in the nuclear envelope, and are expressed at a low level in several nongametogenic adult tissues as well as many cancers. Presence of a binding site for CTCF and its testis-specific paralogue BORIS in the SPANX promoters suggests, by analogy to MAGE-A1 and NY-ESO-1, that their activation in spermatogenesis is mediated by the programmed replacement of CTCF by BORIS. Based on the relative density of CpG, the more extended expression of SPANX-N compared to SPANX-A/D in nongametogenic tissues is likely attributed to differences in promoter methylation. Our findings suggest that the recent duplication of SPANX genes in hominoids was accompanied by different localization of SPANX-N proteins in post-meiotic sperm and additional expression in several nongonadal tissues. This suggests a corresponding functional diversification of SPANX gene families in hominoids. SPANX proteins thus provide unique targets to investigate their roles in the function of spermatozoa, selected malignancies, and for SPANX-N, in other tissues as well. PMID- 17406685 TI - Organometallic molecular materials: self-assembly through hydrogen bonding of an organoplatinum network structure with zeolite-like topology. AB - The complex [Pt(OH)(2)Me(2)(dpa)], dpa = di-2-pyridylamine, undergoes a remarkable form of self-assembly through hydrogen bonding in the solid state; there are cyclic tetramer units formed by head-to-head association between pairs of hydroxyl groups, and cyclic hexamer units formed by head-to-tail association between NH and OH groups; these building blocks then self-assemble to form a complex network structure containing large interconnected cavities, reminiscent of zeolites. PMID- 17406686 TI - A recipe for new organometallic polymers and oligomers? Synthesis and structure of an oligo- and a polymeric arrangement of P-S anions. AB - A route to organometallic polymers and oligomers is described using metal complexes with P/S-ligands as examples. PMID- 17406687 TI - Highly efficient P-N nickel(II) complexes for the dimerisation of ethylene. AB - New P-N ligands featuring a phosphino group and an iminophosphorane moiety were successfully employed in the nickel-catalysed dimerisation of ethylene. PMID- 17406688 TI - Straightforward construction of diarylmethane skeletons via aryne insertion into carbon-carbon sigma-bonds. AB - Two molar amounts of arynes were found to couple with nitriles via carbon-carbon sigma-bond cleavage, assembling diverse diarylmethane skeletons in a straightforward manner. PMID- 17406689 TI - The total synthesis of siphonazole, a structurally unusual bis-oxazole natural product. AB - The first synthesis of the unusual bis-oxazole natural product siphonazole is reported, both oxazole rings being constructed using rhodium carbene chemistry. PMID- 17406690 TI - A bimetallic N-heterocyclic carbene complex featuring a short Cr-Cr distance. AB - The alkylation of a chelating bis(carbene) complex of CrCl(2) yields an unusual bimetallic complex featuring a short Cr-Cr distance, novel ligand coordination, and CH(3) ligand exchange. PMID- 17406691 TI - The hexamethylpentalene dianion and other reagents for organometallic pentalene chemistry. AB - Novel permethylated pentalenide anions are reported which offer exciting new opportunities for the future development of organometallic pentalene chemistry. PMID- 17406692 TI - Bis(permethylpentalene)cerium--another ambiguity in lanthanide oxidation state. AB - Ce(eta(8)-C(8)Me(6))(2) exists in a valency close to Ce(iii) with some Ce(iv) and provides an example of the Kondo effect in a discrete molecule. PMID- 17406693 TI - Activity-based fingerprinting and inhibitor discovery of cysteine proteases in a microarray. AB - A panel of 20 peptide vinyl sulfone probes has been synthesized and used to generate activity-based fingerprinting profiles of cysteine proteases in both gel and microarray-based formats; the inhibitor fingerprints of representative small molecule inhibitors targeted against 4 cysteine proteases were also obtained, in high-throughput, using the same protein microarray platform. PMID- 17406694 TI - Sorption of nitrogen oxides in a nonporous crystal. AB - The uptake of various nitrogen oxides was studied with the well known nonporous p tert-butylcalix[4]arene under ambient conditions. PMID- 17406695 TI - A 'metallic tape' stabilized by an unprecedented (mu5 kappa2,kappa2,kappa2,kappa1,kappa1-) scorpionate binding mode. AB - The Janus scorpionate ligand, tris(mercaptothiadiazolyl)borate, exhibits extraordinary coordination capacity and versatility, binding from two up to five metal cations as demonstrated by its thallium(i) salt, a compound that serves as a model for metal-surface binding. PMID- 17406696 TI - A pcu-type metal-organic framework with spindle [Zn7(OH)8]6+ cluster as secondary building units. AB - The in situ solvothermal reaction of 9,10-dicyanoanthracene and ZnCl(2)/NaN(3) gave the complex, {[Zn(7)(OH)(8)(DTA)(3)].H(2)O}(n) () (DTA(2-) = 9,10 ditetrazolateanthracene), which presents a pcu-type topological framework formed by DTA(2-) bridging unprecedented heptanuclear spindle [Zn(7)(OH)(8)](6+) clusters as SBUs, and exhibits strong luminescent emission with long lifetime. PMID- 17406697 TI - Enantioselective oxidation of O-methyl-N-hydroxylamines using monoamine oxidase N as catalyst. AB - Enantioselective oxidation of racemic O-methyl-N-hydroxycyclohexylethylamine, using a variant of monoamine oxidase N (MAO-N) from Aspergillus niger, yields unreacted (R)-enantiomer (e.e. = 99%) together with the oxime exclusively in the (E)-configuration. PMID- 17406698 TI - Ligand flexibility and framework rearrangement in a new family of porous metal organic frameworks. AB - Ligand flexibility permits framework rearrangement upon evacuation and gas uptake in a new family of porous MOFs. PMID- 17406699 TI - Electropolymeric materials incorporating subsite structures related to iron-only hydrogenase: active ester functionalised poly(pyrroles) for covalent binding of {2Fe3S}-carbonyl/cyanide assemblies. AB - We report the assembly of the first electropolymeric materials incorporating catalytic diiron subsites related to those of the iron-only hydrogenases. PMID- 17406700 TI - Heteronuclear Mn-Ca/Sr complexes, and Ca/Sr EXAFS spectral comparisons with the oxygen-evolving complex of photosystem II. AB - Heterometallic Mn-Ca and Mn-Sr complexes have been prepared and employed as model complexes for Ca and Sr EXAFS spectral comparisons with the Oxygen-Evolving Complex (OEC) of Photosystem II (PS II); these have revealed similarities that support the presence of at least one O atom bridge between the Mn and Ca/Sr in the OEC. PMID- 17406701 TI - Sulfate separation by selective crystallization of a urea-functionalized metal organic framework. AB - Encapsulation of SO(4)(2-) into a Ni coordination framework functionalized with urea anion-binding groups allows selective separation of this strongly hydrophilic anion from a highly competitive aqueous environment. PMID- 17406702 TI - A multi-ion particle sensor. AB - The first sub-micron polyacrylic sensor containing two independent ion-sensing systems is shown, that uses a single excitation wavelength and separates signals by using quantum dot donors to form FRET pairs with other fluorophores. PMID- 17406703 TI - Study on high-efficiency fluorescent microcapsules doped with europium beta diketone complex by LbL self-assembly. AB - Rare-earth beta-diketone complex doped microcapsules with high efficiency fluorescence fabricated by the LbL technique based on electrostatic and charge dipole interactions are reported. PMID- 17406704 TI - Preparation of transparent oxyapatite ceramics by combined use of freeze-drying and spark-plasma sintering. AB - Lanthanum silicate oxyapatites, ion-conducting materials presenting a strong aversion against densification, have been obtained in the form of dense transparent ceramics, by combining the beneficial use of freeze-drying and spark plasma sintering methods. PMID- 17406705 TI - Self-organization of porphyrin units induced by magnetic field during sol-gel polymerization. AB - The use of a magnetic field as a controlling factor during the hydrolysis polycondensation of porphyrin precursors substituted by Si(OR)(3) groups, induces a self-organization of porphyrin moieties due to the stacking of these units in the hybrid material and this study also confirms the effect of the magnetic field in the nano- and micrometric organization during the kinetically controlled polycondensation process. PMID- 17406706 TI - Phosphine containing oligonucleotides for the development of metallodeoxyribozymes. AB - Novel transition metal catalysts based on oligonucleotides can be easily obtained by functionalization of 5-iodouridine with phosphine ligands, resulting in good asymmetric induction in palladium catalyzed allylic nucleophilic substitution. PMID- 17406707 TI - Partially-methylated amyloses as effective hosts for inclusion complex formation with polymeric guests. AB - Partially 2,3-O-methylated amyloses efficiently form inclusion complexes with polytetrahydrofuran and poly(epsilon-caprolactone) by simply mixing them in DMSO H(2)O (1 : 9) solution, in contrast to the case of the parent amylose in which the corresponding inclusion complexes are only slightly formed. PMID- 17406708 TI - Microwave energy: a versatile tool for the biosciences. AB - As the range of techniques for microwave heating has expanded, so have the areas in which it can have a profound impact. Two emerging areas are the application of microwave heating for the synthesis of peptides, peptoids, oligopeptides and carbohydrates and in the field of proteomics. PMID- 17406709 TI - Chiral dipeptide mimics possessing a fluoroolefin moiety: a relevant tool for conformational and medicinal studies. AB - The replacement of the amide bond in a peptide backbone is a promising strategy in peptidomimetic drug research. Over the various amide bond surrogates, the fluoroolefin moiety has been successfully developed as an effective mimic. Today, fluorine-containing compounds account for a large proportion of new active molecules in life sciences. The synthesis of fluoroolefin peptide mimics is not a trivial task and innovative approaches often need to be addressed, in particular for the stereocontrol of the double bond configuration and the chiral centres adjacent to the fluoroalkene. These fluorinated peptidomimetics have been synthesised and evaluated as metabolically stable and/or conformationally constrained analogs of enzyme inhibitors, and as tools for probing the function, structure, and binding process of receptors. PMID- 17406710 TI - Induction of cell death by photodynamic therapy with water-soluble lipid-membrane incorporated [60]fullerene. AB - Intracellular uptake of a lipid-membrane-incorporated C(60) with a cationic surface into HeLa cells was found to induce cell death under visible light irradiation in high efficiency. PMID- 17406711 TI - Asymmetric addition of 1-ethynylcyclohexene to both aromatic and heteroaromatic ketones catalyzed by a chiral Schiff base-zinc complex. AB - The evaluation of a chiral Schiff base ligand in the zinc-catalyzed asymmetric addition of 1-ethynylcyclohexene to both aromatic and heteroaromatic ketones is reported (with up to 83% enantioselectivity and up to 88% isolated yield). PMID- 17406712 TI - Syntheses and anti-HIV activities of (+/-)-norcarbovir and (+/-)-norabacavir. AB - Norcarbovir (1) and norabacavir (2), the desmethylene derivatives of anti-HIV agents carbovir and abacavir, were efficiently synthesized from a common intermediate . Their antitumor and antiviral activities were evaluated and the results indicate norabacavir showed comparable anti-HIV activity to that of abacavir. PMID- 17406713 TI - Efficient synthesis of brussalexin A, a remarkable phytoalexin from Brussels sprouts. AB - The synthesis of brussalexin A, the first phytoalexin containing an allyl thiolcarbamate group, and its selective inhibitory activity against fungal plant pathogens is described. PMID- 17406714 TI - Three-state photochromic switching in a silyl bridged diarylethene dimer. AB - The synthesis and photochemical characterization of bicomponent molecular switches based on covalently tethered dithienylethene photochromes is described. Both photochromic units undergo complete switching between the fully cyclised and fully non-cyclised states despite a significant level of electronic communication between the individual units and their proximity. PMID- 17406715 TI - Highly enantioselective and efficient synthesis of methyl (R)-o-chloromandelate with recombinant E. coli: toward practical and green access to clopidogrel. AB - Methyl (R)-o-chloromandelate ((R)-), which is an intermediate for a platelet aggregation inhibitor named clopidogrel, was obtained in >99% ee by the asymmetric reduction of methyl o-chlorobenzoylformate (2) (up to 1.0 M) with recombinant E. coli overproducing a versatile carbonyl reductase. PMID- 17406716 TI - Opioid ligands with mixed properties from substituted enantiomeric N-phenethyl-5 phenylmorphans. Synthesis of a micro-agonist delta-antagonist and delta-inverse agonists. AB - Enantiomeric N-phenethyl-m-hydroxyphenylmorphans with various substituents in the ortho, meta or para positions of the aromatic ring in the phenethylamine side chain (chloro, hydroxy, methoxy, nitro, methyl), as well as a pyridylethyl and a indolylethyl moiety on the nitrogen atom, were synthesized and their binding affinity to the mu-, delta-, and kappa-opioid receptors was examined. The higher affinity ligands were further examined in the [(35)S]GTPgammaS assay to study their function and efficacy. 3-((1R,5S)-(-)-2-(4-Nitrophenethyl)-2-aza bicyclo[3.3.1]nonan-5-yl)phenol ((-)-) was found to be a mu-agonist and delta antagonist in that functional assay and was about 50 fold more potent than morphine in vivo. 3-((1R,5S)-(-)-2-(4-Chlorophenethyl)-2-aza-bicyclo[3.3.1]nonan 5-yl)phenol ((-)-) and several other ligands displayed inverse agonist activity at the delta-opioid receptor. The absolute configuration of all of the reported compounds was established by chemical conversion of (-)- to 1R,5S-(-)-.HBr. PMID- 17406717 TI - Novel strategies for the synthesis of anthrapyran antibiotics: discovery of a new antitumor agent and total synthesis of (S)-espicufolin. AB - Two high-yielding strategies for the synthesis of 4H-anthra[1,2-b]pyran antibiotics have been developed giving access to novel antitumor agent (ED(50) 1.5 microm) and to (S)-espicufolin (3). A key step for the assembly of the tetracyclic 4H-anthra[1,2-b]pyran-4,7,12-trione skeleton is the nucleophilic addition of an aryl lithium species onto an aldehyde which allows the introduction of either an ynone or 1,3-diketo side chain, serving as precursors for an acid-catalysed cyclisation. PMID- 17406718 TI - Synthesis and radical coupling of pyridine-bridged pi-extended tetrathiafulvalene (TTF)-type donors and push-pull analogues. AB - A new family of pi-extended TTF analogues (3a-c) and D-pi-A chromophores (5a-c), in which the electroactive units (1,3-dithiole rings and 2,2-dicyanovinyl groups) are connected through a pyridine bridge with a meta substitution pattern, is reported. The redox behavior of these compounds has been investigated by cyclic voltammetry and theoretical calculations performed at the B3P86/6-31G** level. Unlike many pi-extended TTF derivatives, the 1,3-dithiole rings in compounds 3a-c do not behave independently and two oxidation processes are observed with an anodic separation ranging from 50 to 150 mV. Calculations show that electrons are equally extracted from both dithiole rings. A biradical structure is predicted for the dication state due to the near-degeneracy of the HOMO and HOMO - 1 orbitals. The presence of both donor (D) and acceptor (A) fragments in conjugates results in irreversible oxidation and reduction processes associated with the 1,3 dithiole ring and with the 2,2-dicyanovinyl moiety, respectively. An electrochemical-chemical-electrochemical (ECE) process takes place for all the compounds reported. The chemical process implies the dimerization of the radical cation for compounds 5 and the oligomerization of the biradical dication for compounds 3. The ECE process therefore generates new neutral dimeric (5) or oligomeric (3) species that incorporate the TTF vinylogue core. PMID- 17406719 TI - Dibenzodiaza-30-crown-10-appended bis(zinc porphyrin) tweezers: synthesis and crown-assisted chiroptical behaviour. AB - In our program for developing chirality manipulation systems, we synthesized bis(zinc porphyrin) 1, with a dibenzodiaza-30-crown-10 as a linker unit. Two structural features were examined. The aza-crown segment exhibited an intermolecular interaction with the zinc(ii) of the porphyrin, capable of causing aggregation to form spherical nanostructures, as inferred by concentration dependency of (1)H NMR as well as scanning electron microscopy (SEM) observation. We also consider the crown-based conformation flexibility, in which accommodated K(+) tunes the porphyrin orientation into the tweezers conformation, assisting chirality induction upon complexation with chiral diamine 2. The circular dichroism (CD) intensity change essentially reached a plateau at a [(1R,2R)-2] : [1] ratio of 2 : 1 for which a 45% enhancement in the amplitude of CD spectra was observed compared to the K(+)-free conditions. Use of the crown linker of is not limited to promoting chirality induction with diamines in the presence of K(+); chiroptical probing of unprotected amino acids (Lys, His, Trp, and Phe) using 1 was attained through liquid (1 in CH(2)Cl(2))-liquid (the amino acids in 1 N KOH) two-phase extraction. The amphiphilic properties of the crown segment, as well as the K(+)-assisted tweezers conformation, make it possible to explore a potent way to develop chirality sensors for amino acids in water. PMID- 17406720 TI - Ruthenium half-sandwich complexes as protein kinase inhibitors: derivatization of the pyridocarbazole pharmacophore ligand. AB - A general route to ruthenium pyridocarbazole half-sandwich complexes is presented and applied to the synthesis of sixteen new compounds, many of which have modulated protein kinase inhibition properties. For example, the incorporation of a fluorine into the pyridine moiety increases the binding affinity for glycogen synthase kinase 3 by almost one order of magnitude. These data are supplemented with cyclic voltammetry experiments and a protein co-crystallographic study. PMID- 17406721 TI - Investigation of the synthetic and mechanistic aspects of the highly stereoselective transformation of alpha-thioamides to alpha-thio-beta chloroacrylamides. AB - Treatment of a series of alpha-thioamides with N-chlorosuccinimide results in efficient transformation to the analogous alpha-thio-beta-chloroacrylamides. The mechanistic pathway has been established through isolation and characterisation of intermediate compounds. The scope of the transformation has been explored-aryl and alkylthio substituents, primary, secondary and tertiary amides can be employed. In most instances, the chloroacrylamides are formed exclusively as the Z-stereoisomer; however, with tertiary propanamides or with amides derived from butanoic or pentanoic acid a mixture of E- and Z-stereoisomers is formed. PMID- 17406722 TI - 2-Oxoalkyl caged oligonucleotides: one-electron reductive activation into emergence of ordinary hybridization property by hypoxic X-irradiation. AB - Ionizing radiation triggers the activation of caged oligodeoxynucleotides (ODNs) with a 2-oxoalkyl leaving group to give the corresponding normal uncaged strands. We designed and synthesized ODNs caged by a 2-oxopropyl group at a given thymine N(3) position (d(oxo)T) to evaluate their one-electron reduction characteristics. Upon hypoxic X-radiolysis in aqueous solution, the caged ODNs released the 2 oxopropyl group to produce the corresponding uncaged ODNs. Digestion by a restriction enzyme Swa I revealed that caged ODN pre-irradiated in hypoxia could form an ordinary duplex with its complementary strand. PMID- 17406723 TI - Facile synthesis of gamma-alkylidenebutenolides. AB - In this paper, a novel route to gamma-alkylidenebutenolides (gamma-AIBs) by way of stereoselective vinylogous aldol reaction of the unactivated butenolide in simple and general conditions is reported. PMID- 17406724 TI - Di-ionizable p-tert-butylcalix[4]arene-1,2-crown-3 ligands in cone and 1,2 alternate conformations: synthesis and metal ion extraction. AB - Novel di-ionizable p-tert-butylcalix[4]arene-1,2-crown-3 ligands in cone and 1,2 alternate conformations were prepared as potential metal ion extractants. Selective bridging of proximal hydroxyl groups of the calix[4]arene platform by a crown-3 polyether unit was achieved under Mitsunobu reaction conditions. In addition to the carboxylic acid group, the acidity tunable N-(X)sulfonyl oxyacetamide functions [OCH(2)C(O)NHSO(2)X] with X group variation from methyl to phenyl to p-nitrophenyl to trifluomethyl were used as the proton-ionizable groups. Conformations and regioselectivities of the new ligands were established by (1)H and (13)C NMR spectroscopy. Competitive solvent extractions of alkali metal cations and of alkaline earth metal cations from aqueous solutions into chloroform were performed, as were single species extractions of lead(ii) and mercury(ii). PMID- 17406725 TI - Solid-emissive fluorophores constructed by a non-planar heteropolycyclic structure with bulky substituents: synthesis and X-ray crystal structures. AB - Novel solid-emissive indeno[1,2-b]benzo[4,5-e]pyran-11-one-type fluorophores having non-planar structures with sterically hindered substituents (R = butyl, phenyl, and thienyl) have been designed and conveniently synthesized. The fluorescence quantum yields of in 1,4-dioxane were (Phi = 0.053) >> (Phi = 0.013) > (Phi = 0.003). On the other hand, the solid-state fluorescence quantum yields of the fluorophores were (Phi = 0.39) > (Phi = 0.15) > (Phi = 0.06). To elucidate the large differences in the quantum yields in solution and in the solid state and among the fluorophores , we performed time-resolved fluorescence spectroscopic measurements, semi-empirical molecular orbital calculations (AM1 and INDO/S), and X-ray crystallographic analyses of . The comparison of the values of the radiative and non-radiative rate constants determined by the time resolved spectroscopic measurements in solution and in the crystalline state supported that non-radiative decay is reduced by restriction of the rotation of the phenyl and thienyl rings in the solid state. In addition, the X-ray crystal structures demonstrated that, in and , the non-planar structure with sterically hindered substituents prevents the fluorophores from forming short pi-pi contacts and produces strong solid-state fluorescence. On the other hand, in the crystal of , the formation of continuous intermolecular CH[dot dot dot]S bonding between neighboring fluorophores was found to increase short pi-pi contacts and reduce the fluorescence intensity. PMID- 17406726 TI - Stereochemistry of Delta4 dehydrogenation catalyzed by an ivy (Hedera helix) Delta9 desaturase homolog. AB - The stereochemistry of palmitoyl-ACP Delta(4) desaturase-mediated dehydrogenation has been examined by tracking the fate of deuterium atoms located on stereospecifically monodeuterated substrates-(4S)- and (4R)-[4-(2)H(1)]-palmitoyl ACP and (5S)- and (5R)-[5-(2)H(1)]-palmitoyl-ACP. It was found that the introduction of the (Z)-double bond between C-4 and C-5 of a palmitoyl substrate occurs with pro-R enantioselectivity-a result which matches that obtained for a closely related homolog-castor stearoyl-ACP Delta(9) desaturase. These data show that despite the difference in regioselectivity between the two enzymes, the stereochemistry of hydrogen removal is conserved. PMID- 17406727 TI - On the stability of 2-aminoselenophene-3-carboxylates: potential dual-acting selenium-containing allosteric enhancers of A1 adenosine receptor binding. AB - Ethyl-2-amino-4,5,6,7-tetrahydro-1-benzoselenophene-3-carboxylate (4), has been prepared as a potential dual-acting selenium-containing allosteric enhancer of adenosine A(1)A receptor binding utilising a modified Gewald reaction. While preliminary testing indicated that 4 is a superior enhancer of A(1)AR binding than its thiophene counterpart, its instability under mildly acidic conditions is cause for concern. X-Ray crystallography, together with DFT calculations, provide evidence that the decomposition of 4 involves the ring-opening of selenophenium ion (12b) followed by the loss of elemental selenium through a radical chain process. PMID- 17406728 TI - An unprecedented tandem 1,3-dipolar cycloaddition-cheletropic elimination: a facial approach to novel push-pull olefins. AB - The interaction of 2-(phenylthiocarbamoyl) imidazolium inner salts with dimethyl acetylenedicarboxylate produced dimethyl 2-(imidazolin-2-ylidene)-3 thioxobutanedioates in moderate to good yields. The process involved a tandem reaction comprising a 1,3-dipolar cycloaddition and an unprecedented cheletropic elimination of the phenyl isonitrile from a 2-phenyliminodihydrothiophene moiety. NMR and X-ray diffraction studies confirmed that the 2-(imidazolin-2-ylidene)-3 thioxobutanedioates are novel push-pull olefins and have potential applications in nonlinear optical materials. PMID- 17406729 TI - Bicyclic proline analogues as organocatalysts for stereoselective aldol reactions: an in silico DFT study. AB - Density functional theory has been employed in investigating the efficiency of a series of bicyclic analogues of proline as stereoselective organocatalysts for the aldol reaction. Three classes of conformationally restricted proline analogues, as part of either a [2.2.1] or [2.1.1] bicyclic framework, have been studied. Transition states for the stereoselective C-C bond formation between enamines derived from [2.2.1] and [2.1.1] bicyclic amino acids and p nitrobenzaldehyde, leading to enantiomeric products, have been identified. Analysis of the transition state geometries revealed that the structural rigidity of catalysts, improved transition state organization as well as other weak interactions influence the relative stabilities of diastereomeric transition states and help contribute to the overall stereoselectivity in the aldol reaction. These bicyclic catalysts are predicted to be substantially more effective in improving the enantiomeric excess than the widely used organocatalyst proline. Enantiomeric excesses in the range 82-95% are predicted for these bicyclic catalysts when a sterically unbiased substrate such as p nitrobenzaldehyde is employed for the asymmetric aldol reaction. More interestingly, introduction of substituents, as simple as a methyl group, at the ortho position of the aryl aldehyde bring about an increase in the enantiomeric excess to values greater than 98%. The reasons behind the vital energy separation between diastereomeric transition states has been rationalized with the help of a number of weak interactions such as intramolecular hydrogen bonding and Coulombic interactions operating on the transition states. These predictions could have wider implications for the rational design of improved organocatalysts for stereoselective carbon-carbon bond-forming reactions. PMID- 17406730 TI - [Predictive value of history and physical examination for the diagnosis of community-acquired pneumonia in adults]. AB - BACKGROUND: Community-acquired pneumonia in adults is a serious health problem in the ambulatory care setting. AIM: To define clinical variables associated with the presence of pneumonia in adult patients presenting with fever or respiratory symptoms to the emergency department. MATERIAL AND METHODS: Prospective study carried out in the emergency department from the Catholic University Hospital in Santiago, Chile. Three hundred twenty-five patients (53+/-22 years) presenting fever or acute respiratory symptoms were included. After obtaining a clinical history and physical examination, the physician established a tentative diagnosis. Subsequently, a definitive diagnosis was made with the chest X rays. RESULTS: Thirty-four percent of the patients had pneumonia. The clinical diagnosis of pneumonia before X-ray examination was variable among emergency physicians (positive likelihood ratio: 1.5-4.8) and showed only moderate sensitivity (79%) and specificity (66%). The clinical variables significantly associated with the presence of pneumonia were: advanced age (over 75 years), cardiovascular disease, fever, chills, sputum production, orthopnea, altered mental status, cyanosis, dullness on percussion, bronchial breath sounds, crackles, any abnormal vital sign (heart rate>or=100 beats/min, respiratory rate>or=20 breaths/min or temperature>or=38 degrees C) and oxygen saturation below 90% breathing air. CONCLUSIONS: Clinical judgment prior to observation of chest X rays had moderate sensitivity and specificity for the diagnosis of pneumonia. There were no individual clinical findings, or combination of findings, that could confirm or exclude the diagnosis of pneumonia for a patient suspected of having this illness. PMID- 17406731 TI - [Extra anatomical revascularization and endovascular stent-grafting for thoracoabdominal aneurysm repair. Report of four cases]. AB - Surgical treatment of thoracoabdominal aneurysms is a big technical challenge with a high rate of complications and mortality. It requires a large exposure and transient interruption of vital organ perfusion during its repair. Endovascular repair is a less invasive alternative available over the last decade. We report four male patients aged 44 to 76 years, with thoracic aortic aneurysms and involvement of visceral aorta, treated with a two stage procedure. During the first stage, a retrograde revascularization of the superior mesenteric and renal arteries from the infrarenal aorta was done, associated in two cases to a concomitant repair of an infrarenal aortic aneurysm. In the second stage, an endovascular graft was placed through the femoral artery, from the segment proximal to the aneurysm to the infrarenal aorta, above the origin of the visceral artery reconstructions, excluding the aneurysm from circulation. In one patient, both stages were concomitant and in three the second stage was delayed. One patient presented a postoperative bleeding that required reintervention without adverse consequences. No patient died, presented paraplegia or deterioration of renal function. After follow up of 6 to 20 months, there is no evidence of aneurysm growth or complications derived from the procedure. PMID- 17406732 TI - [Nasal carriage of Streptococcus pneumoniae in elderly subjects according to vaccination status]. AB - BACKGROUND: S pneumoniae is the main cause of community-acquired pneumonia in the elderly, group that concentrates 95% of deaths. AIM: To assess the prevalence of nasal carriage of S pneumoniae in institutionalized elderly patients. MATERIAL AND METHODS: One hundred eighteen institutionalized subjects aged over 60 years (65 males) were enrolled. Since they were also participating in a controlled protocol related to the immunogenicity of an anti-pneumococcal vaccine, our investigation was also blind and randomized. According to randomization, they received pneumococcal or tetanic vaccine. Nasal swab cultures were taken at the beginning of the trial and two months after vaccination. According to recommended methods, we identified S pneumoniae, the serotypes and their antimicrobial susceptibility. RESULTS: In the first nasal sample, 16% of subjects were positive for S pneumoniae. The second sample was positive in 12%. Of the 33 isolated serotypes, 9.1% demonstrated intermediate resistance to penicillin and 3.3% were resistant to chloramphenicol. CONCLUSIONS: The study demonstrated a greater percentage of colonized patients than in the general population. The isolated serotypes are the same that cause invasive diseases in this age group, according to data of the Institute of Public Health of Chile. There were no differences in the percentage of colonization between subjects vaccinated against S pneumoniae and control groups, after two months of follow up. Isolated strains had a low resistance to penicillin. High level resistance was not observed. PMID- 17406733 TI - [Detection and typification of human papilloma virus in pre cancerous cervical lesions]. AB - BACKGROUND: The association of different genotypes of human papilloma virus (HPV) with cervical cancer is well known. However, there is little information about their association with pre-cancerous lesions. AIM: To assess the frequency of different HPV genotypes in pre cancerous cervical lesions. MATERIAL AND METHODS: A cervical sample was obtained by cytobrush in 15 women with low grade lesions and 40 women with high grade lesions, subjected to conization by loop electrical excision procedure (LEEP). Detection and typification of HPV was done by polymerase chain reaction and restriction fragment length polymorphism. RESULTS: All women were infected with HPV. Eighty five percent of samples were typified. A unique HPV subtype was found in 76% of women. Fourteen percent had an infection with multiple subtypes and in 10%, the viral genotype was not identified. The most common subtypes found were HPV 16, HPV 52 and HPV 53. CONCLUSIONS: There is a high rate of infection with HPV with a high oncogenic risk among these women. PMID- 17406734 TI - [Prevalence of metabolic syndrome in a sample of Chilean children consulting in an obesity clinic]. AB - BACKGROUND: In Chile between 1986 and 1998, the prevalence of obesity has increased from 4.6% to 24% in prepuberal and from 2.3% to 17% in puberal children. AIM: To assess the prevalence of metabolic syndrome (MS) in a sample of obese and overweight Chilean children. PATIENTS AND METHODS: Descriptive study in 489 children (273 females), aged from 6 to 16 years, consulting in an obesity program. Body mass index, Tanner puberal development, waist circumference, blood pressure, fasting blood glucose, insulin and lipid levels were measured. Insulin sensitivity was calculated using the mathematical models QUICKI and HOMA. MS was diagnosed when three or more of the following criteria were met: a waist circumference over percentile 90, a blood pressure over percentile 90, serum triglycerides over 110 mg/dl, HDL cholesterol of less than 40 mg/dl or a fasting blood glucose level over 100 mg/dl. RESULTS: MS was present in 4% of children with overweight and 30% of obese children. No differences in prevalence were observed for sex or puberal development. The most prevalent cardiovascular risk factors were abdominal obesity, present in 76% of the sample and high triclycerides levels in 39%. The less prevalent risk factor was a fasting blood glucose over 100 mg/dl in 3.7%. The risk for MS increased ten fold in children with severe obesity, compared with those with overweight. Those with abdominal obesity had a 17 times higher risk of MS. Basal insulin sensitivity was significantly associated with the number of cardiovascular risk factors and the presence of MS. CONCLUSIONS: In this sample of children, fat distribution and the severity of obesity were strong determinants of MS. Waist circumference had a higher impact on MS than body mass index. PMID- 17406735 TI - [Diagnosis of Helicobacter pylori infection in children based on stool antigen test]. AB - BACKGROUND: During infancy, preventive, diagnostic and therapeutic efforts for Helicobacter pylori infection should be made. AIM: To evaluate non-invasive diagnostic methods such as stool antigen test (HpSA) and serum anti-H pylori antibody detection (IgG e IgA), compared to endoscopy-based invasive methods (histology and urease test) for the diagnosis of Helicobacter pylori infection. PATIENTS AND METHODS: Thirty nine children (aged 3 to 14 years, 20 males) referred for upper gastrointestinal endoscopy, were studied. The gold standard to diagnose Helicobacter pylori infection was defined as a positive invasive diagnostic test (histology and/or urease test). Sensitivity (S), specificity (E) and positive (PPV) and negative (NPV) predictive values were obtained for HpSA and serum antibodies. RESULTS: Ten children (26%) were infected with H pylori. S, E, PPV and NPV for HpSA were 90, 100, 100 and 97%, respectively. The figures for serum IgG were 81, 97, 89 and 93%, respectively and for IgA, 90, 76, 36 and 96%, respectively. CONCLUSIONS: HpSA was sensitive and specific as a clinical and epidemiological tool to evaluate H pylori infection in children. Serology was not as accurate, but IgG had a better performance than IgA. PMID- 17406736 TI - [Etiological study of gynecomastia: results of a prospective study and recommendations]. AB - BACKGROUND: Gynecomastia can be physiological or pathological. A limited study of gynecomastia is recommended during puberty and in the elderly, ages in which gynecomastia is usually considered physiological. Other authors suggest that this condition should be studied when it is painful, rapidly growing, of recent onset, when its diameter is more than 4 cm and when is associated to testicular masses. AIM: To investigate the causes of gynecomastia and to evaluate the above mentioned criteria to exclude pathological conditions. MATERIAL AND METHODS: Prospective study of 117 patients aged 10 to 83 years, consulting for gynecomastia. All were subjected to a standardized study including a clinical examination and measurement of plasma estradiol and testosterone levels. RESULTS: Forty one percent of gynecomastias were considered pathological and the rest, physiological. Among pathological conditions, 18 patients had an endocrine etiology (hypogonadism in ten patients, estrogen secreting tumors in three, hyperestrogenism of unknown etiology in four and peripheral resistance to androgens in one), in 17 it was secondary to medications and in 13 it was secondary to other causes (idiopathic, pesticide exposure, alcoholism, diabetes or re feeding). In 79% of 86 patients of less than 20 years, the condition was physiological and in four of five elderly subjects, it was pathological. Thirty nine percent of pathological gynecomastias lacked the signs and symptoms that according to authors, should prompt a thorough study. CONCLUSIONS: All patients with gynecomastia should be studied with a complete medical history and the measurement of estradiol and testosterone levels. The criteria proposed to conduct minimal studies in gynecomastia, would miss a large volume of pathological conditions. PMID- 17406737 TI - [Effects of folic acid fortification in the rates of malformations at birth in Chile]. AB - BACKGROUND: The effects of folic acid fortification on neural tube defects is well known. Other reports show a beneficial effect of the fortification on orofacial clefts, urinary malformations and defects caused by limb reduction. AIM: To determine the changes in prevalence of congenital malformations after the start of flour folic acid fortification in Chile. MATERIAL AND METHODS: The rates of 22 malformations occurring in the maternity of the University of Chile Clinical Hospital and other Chilean hospitals participating in the Latin American Collaborative Study of Congenital Malformations (ECLAMC) were compared before and after the start of flour folic acid fortification. RESULTS: After the start of folic acid fortification a significant reduction in the rates of anencephalia, spina bifida and diaphragmatic hernia, was observed. The rates of all other malformations remained stable or increased. The rates of all malformations at the University of Chile Clinical Hospital had a steady increase until 2005 and were significantly higher than in the rest of hospitals participating in ECLAMC. CONCLUSIONS: Folic acid fortification was associated with an expected reduction in rates of spina bifida and anencephalia and an unexpected reduction in the rates of diaphragmatic hernia. PMID- 17406738 TI - [Distribution of fasting glucose, insulin, homeostasis model assessment (HOMA) insulin resistance (IR) and HOMA beta cell in children and adolescents from Maracaibo, Venezuela]. AB - BACKGROUND: The raising prevalence of obesity among children increases the risk of insulin resistance and its adverse metabolic consequences. AIM: To determine the distributions of fasting serum glucose, insulin, HOMA IR and HOMA beta cell in a representative sample of children and adolescents from Maracaibo-Venezuela. SUBJECTS AND METHODS: Fasting insulin and glucose were measured in 418 children and adolescents (191 boys and 227 girls) of 7, 9, 11, 13, 15 years of age. HOMA IR and HOMA beta cell were calculated. RESULTS: Insulin levels were lower in 7 and 9 year-old girls and 7 year-old boys compared with 11, 13 and 15 year-old girls and boys. Fasting glucose concentrations were similar in boys and girls. HOMA IR was lower in 7 year-old girls compared to 11, 13 and 15 years-old girls, whereas boys in every age showed similar values. HOMA beta cell was higher in 11 and 13 year-old girls. CONCLUSIONS: Our findings provide useful values to assess insulin resistance and ss-cell functioning in children and adolescents. PMID- 17406739 TI - [Pseudotumoral adenomyomatosis of the gallbladder diagnosed by ultrasound. Report of one case]. AB - Adenomyomatosis of the gallbladder wall is a frequent benign degenerative hyperplasia of unknown etiology. In some cases it looks like a malignant tumor. The diagnosis can be suspected during ultrasound examination. We report a 36 year old male that presented with nonspecific dyspepsia. The ultrasound showed a gallbladder mass with small cyst and echogenic spot inside that suggested the presence of Rokitansky-Ashoff sinuses, seen in adenomyomatosis. The patient was operated and the tumor was excised. The pathological examination of the surgical specimen confirmed the presence of gallbladder adenomyomatosis. PMID- 17406740 TI - [Molecular diagnosis and combined lipid lowering therapy of heterozygous familial hypercholesterolemia. Report of one case]. AB - Heterozygous familial hypercholesterolemia affects one every 400 individuals, is caused by mutations in the LDL receptor gene and is associated with premature coronary artery disease. Nowadays, LDL cholesterol can be efficiently reduced with the new therapies to reduce blood lipids. We report a female patient who consulted in 1975, when she was 46 years old, for severe hypercholesterolemia. In 2003, a sample of leukocyte DNA was obtained and the uncommon 1705+1G>A mutation of the LDL receptor gene was detected. No mutations in the apolipoprotein B gene were found. The patient was treated successfully with simvastatin 80 mg/day and ezetimibe 10 mg/day and LDL cholesterol levels were reduced below 200 mg/dl. PMID- 17406741 TI - [Excess respiratory diseases in children and elderly people in a community of Santiago with high particulate air pollution]. AB - BACKGROUND: The relationship between air pollution and health damage has been sufficiently documented. In station "R" of the air quality monitoring system, located in a community of Metropolitan Santiago (Cerro Navia), the Chilean standard of 150 microg/m3, averaged in 24 hours, for particles with a diameter of 10 micrometers or less (PM10), has been exceeded more days than in the rest of the city stations. AIM: To investigate if the population living near that station has a higher proportion of lower respiratory infections than the Metropolitan Region (MR) as a whole. MATERIAL AND METHODS: An outpatient clinic located near station "R" (Centro Albertz), was implemented as a sentinel center according to UNICEF methodology, used since 1992 by the Acute Respiratory Infections National Program. Daily information was collected between May and December 2004. Monitoring data included total number of consults by children less than 15 years old for lower respiratory tract infections, pneumonia, obstructive bronchitis syndrome in children and by adults over 64 years old for lower airway disease, chronic obstructive pulmonary disease (COPD), and pneumonia. Results were compared with those of the rest of MR. RESULTS: Compared with the MR, children from the sentinel clinic had a significantly higher proportion of consults for obstructive bronchial syndrome (20.1% and 26.4% respectively, in p<0.01) and pneumonia (1.3 and 2.7% respectively, p<0.01). In the elderly, the average consults for lower airway disease were 17% in the sentinel clinic and 12.2% in MR (p<0.04). CONCLUSIONS: Children and elderly subjects at the sentinel clinic had a significantly higher proportion of respiratory infections (pneumonia and obstructive bronchial syndrome in children and lower airway disease in the elderly) as compared to the Metropolitan Region. PMID- 17406742 TI - [The changing epidemiology of viral hepatitis in Chile]. AB - The social and sanitary changes that Chile is experiencing will change the epidemiologic profile of viral hepatitis. Virus A hepatitis will displace to older ages, and immunization plans with specific vaccines should be considered. The real prevalence of hepatitis B may be higher, due to an underreporting of the disease. The education and vaccination of high risk groups should be reinforced. E virus hepatitis requires more research in risk groups and in certain animal species consumed by humans. C virus hepatitis is the greatest challenge as it causes chronic liver disease and is the main cause for liver transplantation. PMID- 17406743 TI - [Trends and main features of Chilean mortality]. AB - BACKGROUND: Mortality data of a community allow to identify its severest diseases and health problems. AIM: To report epidemiologic data on Chilean mortality. MATERIAL AND METHODS: Information about mortality was obtained from the World Health Organization, Pan American Health Organization, the Chilean Institute of Statistics and Ministry of Health. RESULTS: A continuously declining trend in the risk of dying during the last 30 years was observed. Chile has the best situation among South American countries. Infant and general mortality rates fell in 89 and 39%, respectively, during the period 1970-2003. This reduction is observed in all Chilean geographical regions. At the county level, there are 11 counties that duplicate the figure of mortality of the 337 other counties. In the case of infant mortality, there are 91 counties that duplicate the national figure, indicating an important inequity. Mortality is significantly associated with population aging, availability and use of hospital beds. There is a lower level of association with the proportion of population living in urban areas and poverty levels. A projection until 2010 shows and increasing mortality caused by malignant tumors and diabetes and a declining trend for circulatory, respiratory and digestive diseases and accidental deaths. CONCLUSIONS: Mortality has been managed efficiently in Chile but future trends underscore the need for cancer and diabetes prevention and management programs. PMID- 17406744 TI - [The relationship between health care and teaching activities in the context of the Health Care Reform]. AB - The new laws of the health reform in Chile, modified the pre-existent relationship between teaching and health care activities. This relationship allows the training of health care professionals by Universities, in the clinical services of the National System of Health Services that is dependent of the Ministry of Health. The new law established new requirements for specialization and quality of care accreditation. Up to the moment, the relationship between Health Care Services and the Universities was based on harmonic and consensual agreements between both counterparts. With the new law, such relationship becomes more relative and is based on new norms. Therefore, it is of utmost importance to establish a regulated and equilibrated relationship between both institutions, to assure the best possible training of health care professionals, in terms of skills and competences, to adequately satisfy the health needs of the population. PMID- 17406745 TI - [Gene expression profiles for therapeutic decisions in breast cancer]. AB - In the early stages of breast cancer, surgery and radiotherapy arrest the local and regional disease. However 30% of patients will die as a consequence of tumor dissemination. Parameters with a relative predictive value are used to assess candidates to adjuvant therapy. Genomic analysis methods (cDNA microarray) identified a gene expression profile of greater aggressiveness in breast cancer. This knowledge generated the project "Translating Molecular Knowledge Into Early Breast Cancer Management" of the Breast International Group (BIG). This is a consortium of 32 research groups in which Chile participates through the Chilean Cooperative Oncological Research Group (GOCCHI). The project consists in three protocols. In the first, denominated MINDACT (Microarray for Node Negative Disease may Avoid Chemotherapy), 8000 women will be evaluated by gene expression profile and clinical pathological criteria to decide adjuvant therapy. Since a group of patients will not receive therapy, solely based on gene expression profile, GOCCHI is preparing a follow up protocol based on the expression of 22 genes. MINDACT is the first prospective effort to evaluate gene expression profiles in clinical practice. Our participation in this protocol, installs us in the first line of worldwide oncological investigation. PMID- 17406746 TI - [The archeology of San Vincente de Paul Hospital in Santiago, Chile]. AB - The San Vicente de Paul Hospital was the first Clinical Hospital of the University of Chile and was located at the same place of present School of Medicine. The School area contains several old buildings, which are probably remains of the San Vicente de Paul Hospital. After a careful study of the current plans of the Faculty of Medicine of the University of Chile and those of the San Vicente de Paul Hospital, and after checking measurements on the actual site, we were able to demonstrate that two and a half clinical rooms of the original building and some parts of the old laundry still remain intact. At present, these constructions are being used as storerooms, student's union offices, and other activities. We expect that this article may contribute to improve the knowledge of our roots by our own as well as by future generations and that it may inspire our authorities to take care and preserve this important patrimonial remains of our national medicine. PMID- 17406747 TI - [New Chilean law regulates scientific research on human genome and cloning]. PMID- 17406749 TI - [Inappropriate decontamination of gastrointestinal endoscope and acessories: is this the Brazilian reality?]. PMID- 17406750 TI - Microbiologic profile of flexible endoscope disinfection in two Brazilian hospitals. AB - BACKGROUND: [corrected] Endoscopes are routinely used in hospitals and clinics of the world and they can be potential sources of cross-infection when the decontamination process is unsuitable AIM: The routines of flexible endoscope (bronchoscopes, esophagogastroduodenoscopes and colonoscopes) disinfection procedures used in two Brazilian university hospitals were evaluated during a 3 year period METHODS: Aleatory samples from internal channels of endoscopes were collected after patient examination and after cleaning/disinfection procedures RESULTS: A contamination >3 log10 was achieved in samples recovered from endoscopes after patient examination. These samples yielded gram-negative bacilli (n = 142: 56%), gram-positive cocci (n = 43: 17%), yeast cells (n = 43: 17%), and gram-positive bacilli (n = 26: 10%). Approximately, 72 out of 149 samples (48.32%) collected after undergoing the cleaning and disinfection procedures disclosed gram-negative bacilli (n = 55: 61%), gram-positive cocci (n = 21: 23%), gram-positive bacilli (n = 8: 9%) and yeast cells (n = 6: 7%). Esophagogastroduodenoscopes and colonoscopes were the most frequently contaminated devices. Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterobacter spp, Serratia marcescens, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Staphylococcus coagulase negative, Micrococcus luteus, Candida albicans, C. tropicalis, C. glabrata, C. guilliermondii, Bacillus spp and Corynebacterium spp were predominantly identified CONCLUSION: Inappropriate cleaning and low times of disinfection were respectively the major factors associated with the presence of microorganisms in colonoscopes and esophagogastroduodenoscopes. By analyzing the identified germs, hospital disinfection was considered of either intermediate or poor level. After this investigation, both university centers improved their previous protocols for disinfection and conditions for reprocessing endoscopes. PMID- 17406751 TI - [Results of liver transplantation in patients with preoperative diagnosis of hepatocellular carcinoma]. AB - BACKGROUND: [corrected] Hepatocellular carcinoma is the most frequent malignant hepatic tumor in humans, and its association with cirrhosis makes the therapeutic approach still a challenge. Liver transplantation is the treatment of choice for cirrhotic patients with unresectable early hepatocellular carcinoma AIM: To evaluate the post-transplant outcome of a cohort of 15 cirrhotic patients with preoperative diagnosis of unresectable early hepatocellular carcinoma according the Milan criteria who underwent liver transplantation between September 1991 and December 2003 METHODS: We retrospectively reviewed the clinical data from 15 liver transplant recipients and the explanted livers were assessed for the efficacy of preoperative therapy. Patient survival and tumor recurrence were evaluated as primary outcome measures RESULTS: The mean age of the patients was 49.2 +/- 14.3 years and hepatitis C was the etiology of the underlying liver disease in 60%. Preoperative therapy, either chemoembolization or percutaneous ethanol injection, was performed in 12 (86%) patients. Complete necrosis of all tumoral lesions were observed in 5 of 12 patients (44,66%); all others had variable amounts of viable tumor in the explanted liver. Only 4 of the 15 (26.6%) explanted livers had microscopic vascular invasion. The median post-transplant follow-up was 33 months (range: 8-71 months) and no tumor recurrence was detected during this period. The only death was an early event not related to the tumor. The recurrence-free survival rates at 1 and 3 years were 93% CONCLUSION: Liver transplantation has emerged as a good alternative for cirrhotic patients with early hepatocellular carcinoma not amenable to curative resection, offering excellent recurrence-free survival rates. PMID- 17406752 TI - [Steatosis and non-alcoholic steatohepatitis in patients with chronic hepatitis due to hepatitis C virus infection]. AB - BACKGROUND: [corrected] Hepatic steatosis is a common finding in patients with hepatitis C, mainly virus C genotype 3. Steatosis in these cases might be associated with antiviral treatment response and with prognosis of chronic hepatitis. AIMS: To determine the presence of steatosis and non-alcoholic steatohepatitis in chronic hepatitis C and its correlation with genotype and hepatic fibrosis. PATIENTS AND METHODS: One hundred and twenty patients with chronic hepatitis C were retrospectively evaluated; genotype was done in 102 patients. All specimens were stained with hematoxylin-eosin, picrosirius and perls. Staging of hepatitis C was scored by Brazilian Classification and the diagnosis of non-alcoholic steatohepatitis by the American Association for the Study of Liver Diseases criteria RESULTS: Steatosis was found in 65 of 120 cases (54.2%); it was mild in 37/65 (56.9%), moderate in 12/65 (18.5%) and severe in 10/65 (15.4%). In relation to fibrosis, 80 of 120 patients had mild fibrosis F0 F2 (66.6%) and 40 (33.3%) had more advanced fibrosis (F3 or F4). Steatosis was associated with genotype 3 (76.7%) in comparison with other genotypes (49,0%). There were no relationship between steatosis and advanced fibrosis F3/F4 (52,5%) or mild fibrosis (54,4%). Non-alcoholic steatohepatitis was diagnosed in 8/120 cases (6.7%) and was significantly associated with hepatitis C virus genotype 3 and with advanced fibrosis (F3 and F4) CONCLUSIONS: The presence of steatosis and non-alcoholic steatohepatitis in chronic hepatitis C are associated to genotype 3; moreover non-alcoholic steatohepatitis was correlated with more advanced fibrosis. PMID- 17406753 TI - Evaluation of nutritional status of nonhospitalized patients with liver cirrhosis. AB - BACKGROUND: [corrected] Protein-calorie malnutrition is a frequent finding in patients with chronic liver disease, but investigations of nutritional status have been rarely performed in individuals seen at outpatient clinics AIM: To evaluate the nutritional status of patients with alcoholic and nonalcoholic cirrhosis, attended for the first time at a reference outpatient clinic for liver diseases. PATIENTS AND METHODS: A total of 300 consecutive patients attended at the outpatient clinics of a reference center for liver diseases were investigated. Anthropometric evaluation was performed by the usual parameters: triceps skinfold, arm circumference and arm muscle circumference. Biochemical parameters included creatinine/height index, serum albumin and lymphocytes count. The nutritional diagnosis was based on the PCM score proposed by Mendenhall et al. Food intake was retrospectively evaluated using 24-hour dietary recall data. RESULTS: About 71% of the patients studied were chronic alcohol abusers, whereas in 29% cirrhosis was of nonalcoholic etiology. Independently of the disease etiology 75.3% of the patients showed some degree of protein-calorie malnutrition, which was moderate or severe in 38.3% of them. More advanced protein-calorie malnutrition degrees were associated with lower energy and protein intake. The prevalence of moderate or severe protein-calorie malnutrition was higher in patients classified as Child-Pugh C than in patients classified as Child-Pugh A (21% x 58%, respectively). Regarding sexual differences, fat reserves, evaluated by triceps skinfold, were more depleted in females than in males (48.6% x 26.6%) regardless of the etiology of the cirrhosis, whereas muscle reserves (arm muscle circumference) were more depleted in males (43.4% x 13.4%) regardless of the etiology of cirrhosis. In contrast, cirrhosis of alcoholic etiology was determinant in reducing arm muscle circumference in females (20% x 9.1%) CONCLUSIONS: These data highlight the high prevalence of protein-calorie malnutrition occurring early in the natural history of the disease and accompanying functional hepatic deterioration. In addition, attention should be paid to the different gender patterns of response to protein-calorie malnutrition in these patients. PMID- 17406754 TI - Daily interferon induction regimen using different manufactured interferons (alpha-2A or alpha-2B) in combination with ribavirin for treatment of chronic hepatitis C: a prospective randomized study. AB - BACKGROUND: [corrected] Studies on hepatitis C virus kinetics showed that serum levels of interferon fall 48 h after drug administration, when viral load is increasing again. Previously to the availability of pegylated interferon, daily induction therapy with standard interferon was under evaluation. AIMS: To evaluate the safety and efficacy of interferon alpha daily induction regimen in combination with ribavirin. PATIENTS AND METHODS: A randomized trial including 93 patients with chronic hepatitis C was carried out. On satisfying all eligibility criteria, patients were randomly allocated to two different treatment groups: 44 individuals in treatment arm A: IFN 3 MU thrice weekly + ribavirin 1.0-1.2 g daily for 48 weeks (IFN TIW) and 49 individuals in treatment arm B: IFN 3 MU daily + ribavirin 1.0-1.2 g daily for 12 weeks followed by IFN 3 MU thrice weekly + ribavirin 1.0-1.2 g daily, until completion of 48 weeks of therapy (IFN QD). HCV genotyping was obtained in 85 subjects. A negative HCV-RNA 6 months after cessation of therapy was considered a sustained virological response RESULTS: Eighty three patients completed treatment, five dropped out (one from IFN TIW and four from IFN QD) and in five patients therapy was discontinued due to medical request (two from IFN TIW and three from IFN QD). There was no statistically significant difference between groups with respect to therapy interruption. The frequency of cirrhosis was 29%, similar in both groups. In the "intention to treat" analysis the overall sustained virological response was 39.8%. There was no significant difference in sustained virological response rate between both treatment strategies (36.4% IFN TIW vs 42.9% IFN QD). In the 83 patients who finished the trial, sustained virological response was 44.6%. Among subjects with HCV genotype-1, the sustained virological response was 42% (40.9% IFN TIW vs 42.9% IFN QD) and among patients with HCV genotype 2 or 3, the sustained virological response was 55.6% (50% IFN TIW vs 63.6% IFN QD) CONCLUSIONS: Combination therapy had an overall sustained virological response rate of 39.8% ("intention to treat analysis"). There was no difference with respect to sustained virological response rates between patients who used daily induction schedule compared to standard regimen. Adverse events, even more frequent in the daily induction group, did not interfere with the treatment strategies. PMID- 17406755 TI - [Ulcerations in Chagas' megacolon operated at urgency and electively]. AB - BACKGROUND: [corrected] The megacolon is a frequent disease in our emergencie hospital, and approached in the urgency by your complications as fecal impaction, volvulus and perforations. The ulcerations in the megacolons contribute as probable sites of perforations AIM: To compare the frequencies of stercoral ulceration in Chagas' megacolon operated at urgency, by volvulus or fecal impaction, and electively, aiming at a better surgical conduct in the urgency surgery METHODS: It was analyzed 356 anatomy-pathological exams from colon resection of operated patients due to Chagas' megacolon at urgency (102 cases; 29%) and electively (254 cases; 71%), from 1980 to 2000. The surgical urgency indications were attributed to volvulus (71 cases; 69,6%), fecal impaction (25 cases; 24,5%), perforated acute abdomen after rectal catheter or sigmoidoscopy (6 cases; 5,9%). The ulceration frequency was compared in both groups of resections, using chi-square RESULTS: The pathological anatomy - of surgery resection obtained at urgency surgeries, showed 26 cases of ulceration (25,5%) and in electively resections were verified 21 cases of ulceration (8,25%). The difference observed was statistically significant. The comparison among the groups of volvulus; fecal impaction and volvulus with fecal impaction, separately with electively surgery group evidenced significant differences in relation to volvulus and fecal impaction CONCLUSIONS: The higher frequency of ulcerations in the megacolon operated at urgency character emphasizes the needs of immediate resection of sigmoid colon, instead of conservative conduct of simple decompression colostomy, even in exploration laparotomy which the macroscopic examination of sigmoid does not show necrotic signs. This way, should prevent the occurrence of perforation in megacolon at mediate postoperative, with serious results. PMID- 17406756 TI - Bcl-2 expression in rectal cancer. AB - BACKGROUND: [corrected] Proteins involved in apoptosis process seem to play an important role in colorectal carcinogenesis AIM: To determine the prevalence of bcl-2 protein immunohistochemical expression and its relation with clinical and histopathological variables of rectal adenocarcinoma. PATIENTS AND METHODS: One hundred and thirty-two patients operated at "Hospital de Clinicas de Porto Alegre", Porto Alegre, RS, Brazil, between 1988 and 1999 were studied through immunohistochemical reaction using a monoclonal antibody anti-bcl-2 on formalin fixed, paraffin-embedded tissue samples RESULTS: The prevalence of bcl-2 protein was 29.5%. There was a significant increased number of positive bcl-2 cases among women as compared to men. There was no significant association between bcl-2 and age, tumour site, histological grade, mucin production, depth of invasion, lymphatic involvement, distant metastasis or stage, despite a trend showing decreased immunoreactivity to bcl-2 among poorly and moderately differentiated tumours, as well as disseminated disease CONCLUSIONS: Analysis of bcl-2 protein expression in tumour tissues, as well as other oncoproteins, may have a role in predict therapeutic response and prognosis of colorectal cancer. However, the potential use of bcl-2 protein assessment in the clinical set for management of rectal cancer remains to be determined. PMID- 17406757 TI - Prevalence of Helicobacter pylori infection in advanced gastric carcinoma. AB - BACKGROUND: [corrected] There is substantial evidence that infection with Helicobacter pylori plays a role in the development of gastric cancer and that it is rarely found in gastric biopsy of atrophic gastritis and gastric cancer. On advanced gastric tumors, the bacteria can be lost from the stomach. AIMS: To analyze the hypothesis that the prevalence of H.pylori in operated advanced gastric carcinomas and adjacent non-tumor tissues is high, comparing intestinal and diffuse tumors according to Lauren's classification METHODS: A prospective controlled study enrolled 56 patients from "Hospital Universitario", Federal University of Rio Grande do Norte, Natal, RN, Brazil, with advanced gastric cancer, treated from February 2000 to March 2003. Immediately after partial gastrectomy, the resected stomach was opened and several mucosal biopsy samples were taken from the gastric tumor and from the adjacent mucosa within 4 cm distance from the tumor margin. Tissue sections were stained with hematoxylin and eosin. Lauren's classification for gastric cancer was used, to analyse the prevalence of H. pylori in intestinal or diffuse carcinomas assessed by the urease rapid test, IgG by ELISA and Giemsa staining. H. pylori infected patients were treated with omeprazole, clarithromycin and amoxicillin for 7 days. Follow up endoscopy and serology were performed 6 months after treatment to determine successful eradication of H. pylori in non-tumor tissue. Thereafter, follow-up endoscopies were scheduled annually. Chi-square and MacNemar tests with 0.05 significance were used. RESULTS: Thirty-four tumors (60.7%) were intestinal-type and 22 (39.3%) diffuse type carcinomas. In adjacent non-tumor gastric mucosa, chronic gastritis were found in 53 cases (94.6%) and atrophic mucosa in 36 patients (64.3%). All the patients with atrophic mucosa were H. pylori positive. When examined by Giemsa and urease test, H. pylori positive rate in tumor tissue of intestinal type carcinomas was higher than that in diffuse carcinomas. In tumor tissues, 34 (60.7%) H. pylori-positive in gastric carcinomas were detected by Giemsa method. H. pylori was observed in 30 of 56 cases (53.5%) in tissues 4 cm adjacent to tumors. This difference was not significant. Eradication of H. pylori in non-tumor tissue of gastric remnant led to a complete negativity on the 12th postoperative month CONCLUSIONS: The data confirmed the hypothesis of a high prevalence of H. pylori in tumor tissue of gastric advanced carcinomas and in adjacent non-tumor mucosa of operated stomachs. The presence of H. pylori was predominant in the intestinal-type carcinoma. PMID- 17406758 TI - [Functional dyspepsia and depression as an associated factor]. AB - AIM: To assess the association between depression and functional dyspepsia. PATIENTS AND METHODS: Three hundred and forty eight dyspeptic patients were included in a cross-sectional study in the gastroenterology outpatient clinic of a University Hospital in southern Brazil (Pelotas, RS) within a 1-year period (from March, 2001 to March, 2002). The assessment of depression occurred after the diagnosis of dyspepsia. The presence (or not) of depression was verified in both functional and organic dyspeptic patients. In a second moment, the results were compared, an univariate analysis was used to describe the frequencies of the interest variables and a chi-square for the comparison between proportions of the categorical variables. The logistic regression technique was used to establish the odd of functional dyspeptic patients to present depression and to control the effect of other variables in the outcome variable RESULTS: Showed greater prevalence of depression among functional dyspepsia patients (30,4%), when compared to organic dyspepsia patients (11,2%). Women showed greater risk to present functional dyspepsia (OR: 1,74, IC 95%, 1,05-2,89) and in terms of age, the group with ages between 31 to 50 years (OR: 0,28 IC 95%, 0,13-0,54) and 51 to 60 years (OR: 0,41, IC 95%, 0,17-0,96) showed protection effect (subjects within this age groups have minor risk to present functional dyspepsia). After the multivariate analysis depressed patients showed three times greater comorbidity with functional dyspepsia when compared to non-depressed patients (OR 3, 13; IC 95%; 1, 71-5, 74). DISCUSSION: The adjustment for the gender, age and marital status variables confirmed the association between functional dyspepsia and depression. Results point the need to assess the presence of depression in functional dyspepsia patients and to establish specific treatment strategies for these patients. PMID- 17406759 TI - Suprapapillary needle puncture for common bile duct access: laboratory profile. AB - BACKGROUND: [corrected] Biliary cannulation to perform endoscopic retrograde cholangiopancreatography may be difficult due to technical reasons and often is necessary to perform papillotomy, where complications as pancreatitis and perforation may occur AIM: To show minimal complications by a new model of biliary access by means of the suprapapillary needle puncture and its laboratory profile. PATIENTS AND METHODS: After the approval of the protocol by the Scientific Ethics Committee of the institution a free and informed consent was signed by all patients participating in the study. From July 2003 to August 2004, fulfilling the inclusion and exclusion criteria, 30 patients were selected for endoscopic retrograde cholangiopancreatography, using the suprapapillary puncture technique. All patients remained hospitalized, fasting and with basal hydroelectrolytic replacement, were clinically followed up and samples for the determination of serum amylase, lipase and C-RP (C-reactive protein) were collected before and 4 h, 12 h and 24 h after the procedure and reevaluated 60 days after the procedure. Laboratory parameters were submitted to statistical study using analysis of variance for repeated measurements. Multiple comparisons were made based on Wald's statistics RESULTS: The technique was successful in 93.4% (28/30) of the patients. No statistically significant difference regarding to the laboratory profile were observed. Complications related to the technique of papillary puncture occurred in 1/28 patients by not using the guide wire and in 1/28 where mild hemorrhage after dilation of the papillary fistula occurred. Regarding complications related to therapeutic procedures, there were 2/28 retroduodenal perforations, with one (1/30) following unsuccessful puncture and another due to the passage of Dormia's basket through the dilated fistula path. All patients submitted to diagnostic puncture and evaluated 60 days after the procedure presented with the major duodenal papilla of normal aspect. The patients with dilation of the suprapapillary fistula showed the fistula continuing to drain clear bile CONCLUSION: Suprapapillary puncture allows investigative and therapeutic procedures without significant increases in amylase, lipase and C-RP. Patients submitted to diagnostic puncture present complete recovery of the papilla, while dilation of the fistula maintains it pervious later on, but without complications. PMID- 17406760 TI - Growth evaluation in infants with neonatal cholestasis. AB - BACKGROUND: [corrected] Chronic liver diseases in childhood often cause undernutrition and growth failure. To our knowledge, growth parameters in infants with neonatal cholestasis are not available AIM: To evaluate the nutritional status and growth pattern in infants with intrahepatic cholestasis and extrahepatic cholestasis. PATIENTS AND METHODS: One hundred forty-four patients with neonatal cholestasis were followed up at the Pediatric Gastroenterology Service of the Teaching Hospital, State University of Campinas, Campinas, SP, Brazil, in a 23-year period, from 1980 to 2003. The records of these patients were reviewed and patients were classified into two groups, according to their anatomical diagnosis: patients with intrahepatic cholestasis - group 1, and patients with extrahepatic cholestasis - group 2. Records of weight and height measurements were collected at 4 age stages of growth, in the first year of life: 1) from the time of the first medical visit to the age of 4 months (T1); 2) from the 5th to the 7th month (T2); 3) from the 8th to the 10th month (T3); and 4) from the 11th to the 13th month (T4). The weight-by-age and height-by-age Z scores were calculated for each patient at each stage. In order for the patient to be included in the study it was necessary to have the weight and/or height measurements at the 4 stages. Analyses of variance and Tukey's tests were used for statistical analysis. Repeated measurement analyses of variance of the weight by-age Z-score were performed in a 60-patient sample, including 29 patients from group 1 and 31 patients from group 2. The height-by-age data of 33 patients were recorded, 15 from group 1 and 18 from group 2 RESULTS: The mean weight-by-age Z scores of group 1 patients at the 4 age stages were: T1=-1.54; T2=-1.40; T3= 0.94; T4=-0.78. There was a significant difference between T2 X T3 and T1 X T4. The weight-by-age Z-scores for group 2 patients were :T1=-1.04; T2=-1.67; T3= 1.93 and T4=-1.77, with a significant difference between T1 X T2 and T1 X T4. The mean weight-by-age Z-scores also showed a significant difference between group 1 and group 2 at stages T3 and T4. The mean height-by-age Z-scores at the four stages in group 1 were: T1=-1.27; T2=-1.16; T3=-0.92 and T4=-0.22, with a significant difference between T3XT4 and T1XT4. The scores for group 2 patients were: T1=-0.93; T2=-1.89; T3=-2.26 and T4=-2.03, with a significant difference between T1XT2 and T1XT4. The mean height-by-age Z-scores also showed a significant difference between group 1 and group 2 at T3 and T4 CONCLUSION: The weight and height differences between the groups became significant from the 3rd measurement onward, with the most substantial deficit found in the extrahepatic group. In this group, there is evidence that the onset of weight and height deficit occurs between the first and second evaluation stages. PMID- 17406761 TI - [Functional, microbiological and morphological intestinal findings among human immunodeficiency virus infected children]. AB - BACKGROUND: [corrected] Gastrointestinal tract disorders are frequent among human immunodeficiency virus infected children, with important repercussions on nutrition and survival. Most studies related to this subject were restricted to adults, being less investigated the problem in the children. AIMS: To study intestinal digestion, absorption, microbiological and morphological findings among human immunodeficiency virus infected children. MATERIAL AND METHODS: Eleven human immunodeficiency virus infected children under 13 years old, belonging to clinical categories A, B or C, separated in two groups: five patients with current or recent episode of diarrhea and six patients without diarrhea in the last 30 days preceding entering in study. Investigation proposed: microbiological and morphological analysis of small intestine and rectum biopsy; stool exams for bacterium, parasite, rotavirus, Mycobacterium species and Cryptosporidium; D-xylose test RESULTS: All tested subjects (9/11) had low D xylose absorption (8,4 _ 24,4 mg d/L). Small intestinal mucosa histology findings were nonspecific, represented, in majority, of grade I/II enteropathy (6/10). Increased cellular infiltration of the chorion was observed in all specimens. Rectum histology alterations were also nonspecific, with chorion increased cellular infiltration. Mycobacterim avium intracellulare and Cryptosporidium were the solely microorganisms founded, both in stool CONCLUSIONS: Our study demonstrated high prevalence (100%) of intestinal malabsorption among human immunodeficiency virus infected children, despite the occurrence or not of diarrhea. It was not possible to establish relationships between the presence of microorganisms, intestinal malabsorption, intestinal morphologic findings and the occurrence or not of diarrhea. There was no correlation between D-xylose and intensity of villous atrophy. PMID- 17406762 TI - Effect of hyperthermia on experimental acute pancreatitis. AB - BACKGROUND: [corrected] Recent studies indicate that hyperthermia can change inflammatory mechanisms and protect experimental animals from deleterious effects of secretagogue-induced acute pancreatitis AIM: To evaluate the effects of hyperthermia post-treatment on cerulein-induced acute pancreatitis in rats METHODS: Twenty animals were divided in two groups: group I (n = 10), rats with cerulein-induced acute pancreatitis undergone hyperthermia, and group II (n = 10), animals with cerulein-induced acute pancreatitis that were kept normothermic. In all groups, amylase serum levels, histologic damage, vascular permeability and pancreatic water content were assessed. Acute pancreatitis was induced by administration of two cerulein injections (20 mcg/kg). A single dose of Evans' blue dye was administered along with the second dose of cerulein. All animals also received a subcutaneous injection of saline solution. After this process, animals undergone hyperthermia were heated in a cage with two 100 W lamps. Body temperature was increased to 39.5 degrees C and maintained at that level for 45 minutes. Normothermia rats were kept at room temperature in a second cage RESULTS: Control animals had typical edema, serum amylase activity and morphologic changes of this acute pancreatitis model. Hyperthermia post-treatment ameliorated the pancreatic edema, whereas the histologic damage and the serum amylase level remained unchanged CONCLUSIONS: The findings suggest a beneficial effect of the thermal stress on inflammatory edema in experimental acute pancreatitis. PMID- 17406763 TI - [Allogenic islet transplantation on the rat liver after allogenic nonparenchymal cells injection in the thymus]. AB - BACKGROUND: [corrected] The major indication for pancreas or islet transplantation is diabetes mellitus type I. This process has to supply the insulin necessity keeping glucose under control AIM: We studied allogenic islet transplantation on the rat liver, Wistar (RT1u) to Lewis (RT1(1)) as a recipient. Control group (n = 8) and nonparenchymal cell group (n = 8) respectively with injection of Hanks solution and nonparenchymal cells in the thymus before islet transplantation. MATERIAL AND METHODS: With the method of isolation and purification of the islets we obtained both in the control group 3.637 +/-783,3 islets with purity of 85 +/- 3,52% and nonparenchymal cell group 3.270 +/- 770 islets with purity of 84,25 +/- 2,76%. The nonparenchymal cells were retrieved from the liver and we obtained 2 x 106 cells. Diabetes was induced by i.v. streptozotocin RESULTS: Control group the transplantation of 3.637 +/- 783,3 islets in the rat liver normalized glucose test, 7,21 +/- 0,57 mmol/L in the 2nd postoperative day. Acute rejection came in the 6th postoperative day with significantly increase of glucose test in nonparenchymal cell group, the transplantation of 3.270 +/- 770 islets in the rat liver, almost normalized the glucose test was 17,95 +/- 5,33 mmol/L in the 2nd postoperative day. From the 4th postoperative day to 10th postoperative day. The glucose test increase significantly showing an early acute rejection CONCLUSION: The injection of nonparenchymal cells in the thymus before allogenic islet transplantation in the rat liver lead to an early acute rejection. PMID- 17406764 TI - [Endoscopic treatment of esophageal varices, using pretied loop made with polyamide thread]. AB - BACKGROUND: [corrected] The elastic band ligation is the method of choice for treatment of esophageal varices. The action mechanism is a mechanical varices compression with thromboses. Based on this concept we developed a ligature method using pretied loop made with polyamide thread for the treatment of esophageal varices. OBJECTIVE: The present study describes and evaluates the feasibility of the treatment of esophageal varices by the ligature method using pretied loop made with polyamide thread and analyzes the local changes of the ligations and the results, concerning safety, efficiency and complications of this procedure. PATIENTS AND METHODS: Between March, 1998 and May, 2000, 58 patients with esophageal varices were treated with pretied loop, made with polyamide thread (26 patients with schistosomiasis, 11 with alcoholic cirrhosis, 9 with hepatitis C, 5 with hepatitis B, 4 of unknown etiology, 2 with hepatitis B and C, and one with Budd-Chiari syndrome/ 42 men and 16 women/ average age of 47,67 +/- 13,12 years, range 16-74). A plastic tube was attached to the endoscope tip featured as an accessory working channel, allowing the pretied loop made with polyamide thread to be conducted to the esophagus facilitated by a flexible metallic tube, to perform the esophageal varices ligature. A total of 506 ligatures were done, distributed in 223 sections (average of 2,26 +/- 1,08 ligature, varying from 1 to 6 per section). The sessions were perform with the interval of 15 days, until the complete eradication of the esophageal varices. The ecoendoscopy was used as a complementary method to evaluate the varices eradication RESULTS: The esophageal varices were treated successfully in all patients. The complete eradication of varices was achieved in 47 (81,03%) patients. In 37 (63,79%) patients the ligatures resulted in pseudopolyps. It was not identified systemic complications or obits. The ecoendoscopy showed thrombosis in the pseudopolyps of 10 patients. The follow-up period was from 4 to 32 months and recurrence of the esophageal varices was observed in 9 (15,51%) patients. The average cost of each ligature was estimated in US$ 1,00. CONCLUSION: The study indicates that endoscopic treatment using polyamide thread ligature is a safe, efficient, accessible and low cost method for treatment of esophageal varices treatment, demonstrating a new aspect, that is the obliteration of varices without necrosis, by formation of pseudopolyps. PMID- 17406765 TI - [Gastroesophageal reflux disease: an overview]. AB - BACKGROUND: Gastroesophageal reflux disease, considered one of the most common digestive diseases in western countries, has been very much studied. The great number of publications in medical literature based upon this issue must be pointed out. However, some of its aspects remain controversial. AIMS: To emphasize important topics of the disease and to present a review on the theme. The following topics were reviewed: defi nition, prevalence, clinical complaints, indications and limitations of the major diagnostic methods and medical, surgical and endoscopic treatments. MATERIAL AND METHODS: We searched, by internet: selected review articles, consensus, guidelines and systematic reviews with meta analysis, published in the last 5 years (from 2000 to 2005), in the following data bases: Cochrane Central Register of Controlled Trials (The Cochrane Library) and PubMed. Among lots of papers which were found, we selected nine systematic reviews with meta-analysis, fi ve review articles, fi ve guidelines and four consensus, that dealt with the topics we had decided to review. It must be pointed out that, as it has been observed in our bibliographical references, not only the publications found were taken into account; other relevant ones (some of them published previously to the analyzed time period) were considered in the composition of this present paper. CONCLUSIONS: Among the 12 conclusions presented, two have to be emphasized: 1. the esophageal multichannel intraluminal impedance has largely contributed to the better knowledge of the gastroesophageal reflux disease and it suggests, when associated with pHmetry (esophageal impedance-pHmetry), as a new gold standard to the gastroesophageal reflux diagnosis and 2: among the gastroesophageal reflux disease symptoms some of them may be considered acid dependents and can usually get good responses to the proton pump inhibitors. Nevertheless, there are also some symptoms which are more dependent on the physical presence of the reflux rather than its acidity. Such symptoms, which do not depend so much on the acidity of the reflux, are not well controlled by the proton pump inhibitors. PMID- 17406766 TI - Polymorphisms in the vitamin D receptor gene are associated with muscle strength in men and women. AB - INTRODUCTION: Vitamin D receptor (VDR) polymorphisms have been associated with fracture risk and muscle strength, although evidence for the latter is limited and conflicting. METHODS: BsmI, TaqI and FokI VDR polymorphisms were genotyped in 253 men (54.9 +/- 10.2 yr) and 240 women (41.5 +/- 13.2 yr). Haplotypes were constructed for BsmI and TaqI. Handgrip, isometric (at 60 degrees , 120 degrees and 180 degrees joint angle) and eccentric torques (60 degrees /s) of knee extension and flexion were analysed using AN(C)OVA. Torque-velocity curves were constructed for concentric torques at 60 degrees /s, 180 degrees /s and 240 degrees /s and analysed using multivariate AN(C)OVA. Age, height and fat-free mass were included as covariates. RESULTS: Quadriceps isometric and concentric strength were higher in female f/f homozygotes compared to F allele carriers. Adjustment for confounding factors rendered results for quadriceps isometric strength at 120 degrees non-significant. No significant association was found with BsmI-TaqI haplotype in women. In contrast, male Bt/Bt homozygotes had higher isometric quadriceps strength at 150 degrees and higher concentric quadriceps strength than bT allele carriers without and with adjustment for confounding factors. No association was observed with FokI in men. In both genders, no interaction effect was present between BsmI-TaqI haplotype and FokI. CONCLUSIONS: Different VDR gene polymorphisms are associated with quadriceps strength in men and women. PMID- 17406767 TI - Serum from postmenopausal women directs differentiation of human clonal osteoprogenitor cells from an osteoblastic toward an adipocytic phenotype. AB - A consistent observation in osteoporosis is bone volume reduction accompanied by increased marrow adipose tissue. No single cause linking the two phenomena has yet been identified. In a human progenitor cell clone (hOP 7) derived from bone marrow, however, we have demonstrated that rabbit serum can direct differentiation away from an osteoblast lineage to one of adipocytes. We now report on whether human serum has a similar effect. Serum was collected from 10 pre- and 10 postmenopausal women and from the 10 postmenopausal women before and following 6-week hormone replacement therapy (HRT). hOP 7 cells were cultured with the various sera, and after 7-14 days adipocytogenesis was determined by oil red O staining and lipoprotein lipase (LPL) and glycerol 3-phosphate dehydrogenase (G3PDH) expression. Incubation with 10% premenopausal serum led to labeling of 10.9% of cells (P < 0.05) with oil red O, whereas application of 10% postmenopausal serum led to a much larger effect, 43.5% labeling (P < 0.001 with respect to premenopausal serum). Oil red O positivity was accompanied by loss of type I collagen expression and increased LPL and G3PDH expression. HRT did not reverse the adipocytogenic effect of postmenopausal serum. In conclusion, serum from postmenopausal women contains factors that steer hOP 7 bone progenitor cells toward an adipocytic phenotype, irrespective of HRT. The study suggests a role for serum factors in the development of fatty marrow in postmenopausal osteoporosis. PMID- 17406768 TI - Estrogen metabolism modulates bone density in men. AB - Estrogen is a critical hormone for bone homeostasis in men, but no information is available on the role of estrogen metabolism among men. The aim of this study was to evaluate the effect of estrogen hydroxylation on male bone mineral density (BMD). Participants consisted of 61 healthy Caucasian males (mean age 66.6 +/- 1.0 years). Urinary estrogen metabolites were measured by enzyme-linked immunosorbent assay, serum estradiol by ultrasensitive radioimmunoassay, sex hormone binding globulin by radioimmunoassay, and BMD of the lumbar spine and the proximal femur by dual-energy X-ray absorptiometry. Active estrogen metabolites, 16alpha-hydroxyestrone (16alphaOHE(1)) and estriol (E(3)), positively correlated with adjusted BMD in all regions of the proximal femur (all P < 0.05) but not at the lumbar spine, and those in the highest tertile of urinary 16alphaOHE(1 )had the highest BMD. Free estradiol index (FEI) also positively correlated with BMD of the total hip, femoral neck, and intertrochanter (all P < 0.05), while there was no correlation between BMD with inactive metabolites (2-hydroxyestrone and 2 methoxyestrone) and serum testosterone. Multiple regression analysis showed 16alphaOHE(1), FEI, and body mass index are important independent predictors of BMD in all regions of the proximal femur. Estrogen metabolism may modulate BMD in men. Increased urinary 16alphaOHE(1) and E(3) levels are associated with high BMD at the proximal femur, and 16alphaOHE(1) appears to be a major determinant of BMD among the metabolites evaluated. PMID- 17406769 TI - Exocytotic process as a novel model for mineralization by osteoblasts in vitro and in vivo determined by electron microscopic analysis. AB - The process of biomineralization has been examined during osteoblastic differentiation of bone marrow stroma cells (BMSCs) from embryonic chick in culture and in periosteum itself by a number of different techniques including transmission and scanning electron microscopy. In cell culture of BMSCs at days 20-25, crystals were accumulated extracellularly in the collagen matrix, resulting in large plate-like crystallites and noncollagen associated on the culture disk surface. In contrast, up to days 10-18, mainly intracellular mineralization was visible by numerous needle-like crystal structures in the cell cytoplasm and in vacuoles. After 20-30 days, the crystal content of these vacuoles is released, most probably by membrane fusion to the outside of the cells. Energy-dispersive X-ray analysis (EDX), electron spectroscopic imaging, and electron energy loss spectroscopy demonstrated that Ca, O, and P are located in the intra- and extracellular needle-like crystals. From EDX spectra a Ca/P ratio of 1.3 was estimated for the intracellular structures and a Ca/P ratio of 1.5, for the extracellular material (for comparison, the Ca/P ratio in tibiae is 1.6). X-ray diffraction and quantitative infrared spectral analyses also demonstrated an increase of crystalline bone apatite along the mineralization process. In addition to the finding in vitro, the presence of intracellular needle-like crystals in vacuoles could be demonstrated in vivo in osteoblastic cells of the periosteum in tibia of day 11. The results are in favor of a novel model for mineralization by osteoblasts, in which amorphous Ca/P material is directly secreted via an exocytotic process from vacuoles of the osteoblast, deposited extracellularly, propagated into the collagen fibril matrix, and matured to hydroxyapatite. PMID- 17406770 TI - Improvement of bone quality in gonad-intact middle-aged male rats by long-chain n 3 polyunsaturated fatty acid. AB - The effect of long-chain n-3 polyunsaturated fatty acid (PUFA) on bone measurements was evaluated in gonad-intact middle-aged male rats. Seven rats were killed on day 0 of dietary intervention to determine bone parameters at baseline. Experimental rats (7/group) were fed one of the following lipid treatments (g/kg diet): 167 g safflower oil + 33 g menhaden oil (N6+N3 diet, control), 200 g safflower oil (N6 diet), or 190 menhaden oil + 10 g corn oil (N3 diet). After 20 weeks of dietary treatment, all groups had lower values for peak load and ultimate stiffness in femurs compared to baseline values. Rats fed the N3 diet had the highest values for peak load, ultimate stiffness, and Young's modulus compared with those fed the N6 and control diets. Compared to baseline, all dietary treatment groups had significantly lower values for trabecular thickness and number in proximal tibia but higher values for trabecular separation and formation rate in proximal tibia and endocortical bone formation rate in tibial shaft. Compared with the control group, rats fed the N3 diet had lower values for formation rate, osteoclast number, and eroded surface in proximal tibia but higher values for periosteal mineral apposition and formation rates in tibia shaft. These findings indicate that a diet rich in long-chain n-3 PUFA mitigate aging-induced loss of bone integrity in intact middle-aged male rats through reducing bone turnover rate by suppressing both bone formation and resorption as a result of a larger net bone volume and modulating endocortical and cancellous bone compartments. PMID- 17406771 TI - Effects of the inoculation of Burkholderia vietnamensis and related endophytic diazotrophic bacteria on grain yield of rice. AB - During a survey of endophytic diazotrophic bacteria associated with different rice varieties in Tamilnadu, some "endophytes" were obtained. Thirteen bacterial isolates from surface-sterilized roots and shoots were obtained in pure culture, which produced indole acetic acid (IAA) and reduced acetylene to ethylene. Polymerase chain reaction (PCR) amplification confirmed the presence of nif-H gene in all the isolates. Morphological, biochemical, and molecular characteristics indicated that all of them belonged to the genus Burkholderia One of them, MGK3, was consistently more active in reducing acetylene, and 16S rDNA sequences of isolate MGK3 confirmed its identification as Burkholderia vietnamiensis. Colonization of rice root was confirmed by strain MGK3 marked with gusA gene. The inoculated roots showed a blue color, which was most intense at the points of lateral root emergence and at the root tip. Transverse sections of roots, 15 days after inoculation, revealed beta-glucuronidase (GUS) activity within many of the cortical intercellular spaces next to the stele and within the aerenchyma. Nitrogen fixation was quantified by using (15)N isotope dilution method with two different cultivars grown in pot and field experiments. Higher nitrogen fixation was observed in variety Ponni than in ADT-43, where nearly 42% (field) and 40% (pot) of the nitrogen was derived from the atmosphere (% Ndfa). Isolate MGK3 was used to inoculate rice seedlings in a comparison with four other diazotrophs, viz., Gluconacetobacter diazotrophicus LMG7603, Herbaspirillum seropedicae LMG6513, Azospirillum lipoferum 4B LMG4348, and B. vietnamiensis LMG10929. They were used to conduct two pot and four field inoculation experiments. MGK3 alone, and combined with other diazotrophs, performed best under both pot and field conditions: combined inoculation produced yield increases between 9.5 and 23.6%, while MGK3 alone increased yield by 5.6 to 12.16% over the uninoculated control treatment. PMID- 17406772 TI - Genetic diversity of Acacia seyal Del. rhizobial populations indigenous to Senegalese soils in relation to salinity and pH of the sampling sites. AB - The occurrence and the distribution of rhizobial populations naturally associated to Acacia seyal Del. were characterized in 42 soils from Senegal. The diversity of rhizobial genotypes, as characterized by polymerase chain reaction restriction fragment length polymorphism (RFLP) analysis of 16S-23S rDNA, performed on DNA extracted from 138 nodules resulted in 15 clusters. Results indicated the widespread occurrence of compatible rhizobia associated to A. seyal in various ecogeographic areas. However, the clustering of rhizobial populations based on intergenic spacer (IGS) RFLP profiles did not reflect their geographic origin. Four genera were discriminated on the basis of 16S rRNA gene sequences of the strains representative for the IGS-RFLP profiles. The majority of rhizobia associated to A. seyal were affiliated to Mesorhizobium and Sinorhizobium 64 and 29%, respectively, of the different IGS-RFLP profiles. Our results demonstrate the coexistence inside the nodule of plant-pathogenic non-N(2)-fixing Agrobacterium and Burkholderia strains, which induced the formation of ineffective nodules, with symbiotic rhizobia. Nodulation was recorded in saline soils and/or at low pH values or in alkaline soils, suggesting adaptability of natural rhizobial populations to major ecological environmental stress and their ability to establish symbiotic associations within these soil environments. These results contribute to the progressing research efforts to uncover the biodiversity of rhizobia and to improve nitrogen fixation in agroforestry systems in sub-Saharan Africa. PMID- 17406773 TI - Environmental stress determines the quality of bacterial lysate and its utilization efficiency in a simple microbial loop. AB - Heterotrophic bacteria provide the critical link in the microbial loop by converting dissolved organic matter (DOM) into particulate form. In this study, DOM was prepared from recently isolated estuarine bacterial strain Vibrio sp. (DSM14379) grown at different salinities [0.2%, 0.5%, 3%, 5%, or 10% (w/v)], washed, concentrated, and lysed by autoclaving. The corresponding lysate containing media were designated LM(0.2), LM(0.5), LM(3), LM(5), and LM(10). Vibrio sp. cells grown at different salinities had similar C/N/P ratios, but different C/S ratios, different trace element composition, and different 2D gel electrophoresis protein profiles. Pseudoalteromonas sp. (DSM06238) isolated from a similar environment was able to grow on all lysates, and its biomass production was dependent on lysate type. The highest growth rate and biomass production of Pseudoalteromonas sp. at saturation lysate concentrations were observed in LM(3). The biomass production at saturation lysate concentrations was about 3-fold higher as compared to LM(0.2) and LM(10). The initial respiration rate, intracellular adenosine triphosphate (ATP) levels, and (3)H-Leu and (3)H-TdR incorporation rates were lowest in LM(3). On the other hand, in LM(0.2) or LM(10) lysates the situation was reversed, the growth rates and biomass production were lowest, whereas (3)H-Leu and (3)H-TdR incorporation, respiration rates, as well as ATP levels, were highest. These results imply uncoupling of catabolism from growth in either high- or low-salinity lysates. The results also suggest that differences in organic carbon quality generated during Vibrio sp. growth at different NaCl concentrations were propagated through the simple microbial loop, which may have important ecological implications for higher trophic levels that depend on microbial grazing. PMID- 17406774 TI - Metabolic activity and phylogenetic diversity of reed (Phragmites australis) periphyton bacterial communities in a hungarian shallow soda lake. AB - In the present study, the species composition and potential metabolic activities of bacterial communities of reed Phragmites australis (Cav.) (Trin. ex Steudel) periphyton from Lake Velencei were studied by cultivation-based and metabolic fingerprinting methods. Serially diluted spring biofilm samples were used to test the community-level physiological profiling (CLPP) using BIOLOG microplates, and for plating onto different media. On the basis of their morphological, biochemical, and physiological test results, 173 strains were clustered by numerical analysis. Representatives of amplified ribosomal DNA restriction analysis (ARDRA) groups were identified by their 16S rDNA sequence comparison. Based on the results of the CLPP investigations, regional differences were detected among the utilized substrate numbers and types, parallel with the increase in incubation time. The phenotypic test results of the strains showed considerable variability with respect to the sampling sites and the media used for cultivation. The most frequently isolated strains were identified as members of genera Agrobacterium, Pseudomonas (P. anguilliseptica, P. marginalis, P. alcaligenes, P. fragi) with aerobic or facultative anaerobic respiratory metabolism, and the species Aeromonas sobria and A. veronii with strong facultative fermentative metabolism. Other strains were identified as Gram positive Arthrobacter, Bacillus, and Kocuria species. The rarely isolated strains were members of beta-Proteobacteria (Acidovorax, Delftia, Hydrogenophaga, and Rhodoferax), gamma-Proteobacteria (Psychrobacter and Shewanella), low G + C Gram positives (Brevibacillus, Paenibacillus, and Exiguobacterium) and high G + C Gram positives (Aureobacterium and Microbacterium). PMID- 17406775 TI - MiCA: a web-based tool for the analysis of microbial communities based on terminal-restriction fragment length polymorphisms of 16S and 18S rRNA genes. AB - A web-based resource, Microbial Community Analysis (MiCA), has been developed to facilitate studies on microbial community ecology that use analyses of terminal restriction fragment length polymorphisms (T-RFLP) of 16S and 18S rRNA genes. MiCA provides an intuitive web interface to access two specialized programs and a specially formatted database of 16S ribosomal RNA sequences. The first program performs virtual polymerase chain reaction (PCR) amplification of rRNA genes and restriction of the amplicons using primer sequences and restriction enzymes chosen by the user. This program, in silico PCR and Restriction (ISPaR), uses a binary encoding of DNA sequences to rapidly scan large numbers of sequences in databases searching for primer annealing and restriction sites while permitting the user to specify the number of mismatches in primer sequences. ISPaR supports multiple digests with up to three enzymes. The number of base pairs between the 5' and 3' primers and the proximal restriction sites can be reported, printed, or exported in various formats. The second program, APLAUS, infers a plausible community structure(s) based on T-RFLP data supplied by a user. APLAUS estimates the relative abundances of populations and reports a listing of phylotypes that are consistent with the empirical data. MiCA is accessible at http://mica.ibest.uidaho.edu/. PMID- 17406776 TI - Efficacy and safety of 1,000 mg effervescent aspirin: individual patient data meta-analysis of three trials in migraine headache and migraine accompanying symptoms. AB - Migraine is often associated with health consequences including impaired quality of life, and the cost of treating migraine headaches places a significant financial burden on patients who suffer from migraines. Nonsteroidal anti inflammatory drugs (NSAIDs) and triptans are commonly used for the treatment of acute migraine attacks. Aspirin is widely accepted as a treatment option for migraine pain relief and could provide an alternative not only for treatment of moderate migraine attacks, but also for severe migraine attacks. The efficacy and safety of 1,000 mg effervescent aspirin (eASA) was evaluated in comparison to 50 mg sumatriptan and placebo in an individual patient data meta-analysis of three randomized, placebo-controlled, single- dose migraine trials. Pain-relief at 2 h, pain-free at 2 h and sustained pain-free up to 24 h were calculated. For eASA, the response rates were 51.5 % (95 % CI: 46.6-56.5 %), 27.1 % (95 % CI: 22.6-31.4 %), and 23.5 % (95 % CI: 19.3-27.7 %). For sumatriptan, the response rates were 46.6 % (95% CI: 40.0-53.2 %), 29% (95 % CI: 23.0-34.9 %), and 22.2 % (95 % CI: 16.7-27.6 %). The corresponding rates for placebo were 33.9 % (95% CI: 29.1-38.6 %), 15.1 % (95 % CI: 11.5-18.7 %), and 14.6 % (95 % CI: 11.0-18.1 %). The treatment effect of eASA and sumatriptan were significantly different from placebo (p < 0.001), but differences between eASA and sumatriptan were not significant. The remission of accompanying symptoms and the subgroup analyses of patients with moderate or severe migraine pain at baseline revealed no significant differences between eASA and sumatriptan. Safety was evaluated based on the frequency of reported adverse events, and treatment with eASA was associated with lower incidence of adverse events than was with sumatriptan. This individual patient data meta-analysis provided evidence that eASA 1,000 mg is as effective as sumatriptan 50mg for the treatment of acute migraine attacks and has a better side effect profile. This is also true for patients with moderate as well as severe headache at baseline. Patients therefore should be advised to use eASA first for migraine attacks and use a triptan in case of no response. PMID- 17406777 TI - Can MR measurement of intracranial hydrodynamics and compliance differentiate which patient with idiopathic normal pressure hydrocephalus will improve following shunt insertion? AB - BACKGROUND: Between 10 and 90% of patients with normal pressure hydrocephalus (NPH) treated with a shunt will improve but they risk significant morbidity/mortality from this procedure. NPH is treated hydrodynamically and it has been assumed that a hydrodynamic difference must exist to differentiate which patient will respond. The purpose of this study is to see whether MRI hydrodynamics can differentiate which patients will improve post shunting. METHOD: Thirty-two patients with NPH underwent MRI with flow quantification measuring the degree of ventricular enlargement, sulcal compression, white matter disease, total blood inflow, sagittal sinus outflow, aqueduct stroke volume, relative compliance ratio and arteriovenous delay. Patients were followed up after shunt insertion to gauge the degree of improvement and were compared with 12 age-matched controls and 12 patients with Alzheimer's disease. FINDINGS: 63% of patients improved with insertion. The responders were identical to the non responders in all variables. The NPH patients were significantly different to the controls (e.g. Total blood inflow reduced 20%, sagittal sinus outflow reduced 35%, aqueduct stroke volume increased 210%, relative compliance ratio reduced 60% and arteriovenous delay reduced 57% with p = 0.007, 0.03, 0.04, 0.0002 and 0.0003 respectively. The patient's with Alzheimer's disease values were midway between the NPH and control patients. CONCLUSIONS: Significant hydrodynamic differences were noted between NPH and controls but these were unable to differentiate the responders from non-responders. The hydrodynamics of Alzheimer's disease makes exclusion of comorbidity from this disease difficult. PMID- 17406778 TI - Double phakomatosis; neurofibromatosis type-1 and tuberous sclerosis. AB - Neurocutaneous syndromes represent some of the most common inherited disorders of the nervous system. Neurofibromatosis type-1 (NF-1) and tuberous sclerosis are well described. Yet, the presentation of both syndromes in the same patient is quite rare. We performed a thorough review of the literature of such double phakomatosis including pattern of inheritance. Eleven cases were reported in the literature. In addition we report a young patient who presented with clinical picture suggestive of both NF-1 and tuberous sclerosis, and present a radiographic and histopathological description of the case. PMID- 17406779 TI - Foraminal L5-S1 disc herniation and conus medullaris syndrome: a vascular etiology? AB - A 56-year-old man presented with a conus medullaris syndrome lumbar magnetic resonance imaging showed only a right foraminal disc herniation at the L5-S1 level. Operative removal of the protrusion was followed by prompt neurological improvement. We postulate that the discrepancy between extent of compression shown by imaging and the neurological findings may reflect vascular compression of the artery of Desproges-Gotteron, an anatomical variant. PMID- 17406780 TI - Early post-operative ACTH and cortisol as predictors of remission in Cushing's disease. AB - AIM: To study the value of early (24 h) post-operative ACTH and serum cortisol as predictors of remission after transsphenoidal surgery in Cushing's disease. METHODS: We prospectively studied 44 patients who underwent transsphenoidal surgery for Cushing's disease between 1997 and 2005. The mean follow-up period of patients after surgery was 49 months (19-102 months). The predictive value of clinical characteristics, pre-operative hormonal studies, radiological, surgical and histological findings, and post-operative hormonal studies were analysed. For the post-operative hormonal study plasma ACTH and serum cortisol were determined at 8.00 a.m. the day after surgery. RESULTS: After surgery, Cushing's disease remitted in 39 patients (89%) and persisted in 5 patients (11%). Three patients relapsed during the follow-up period. Only three study variables were predictive of persistence of Cushing's disease after surgery: the non identification of the adenoma in histology (an adenoma was found in 87% of the patients in remission, and in 20% of treatment failures, p = 0.01), the early post-operative plasma ACTH (patients in remission: 2 pmol/L (1.1-10.8 pmol/L), treatment failures: 8.2 pmol/L (1.1-12 pmol/L), p = 0.019), and the early post-operative serum cortisol (patients in remission: 128.4 nmol/L (27.6-4644 nmol/L), treatment failures: 797 nmol/L (606-1037 nmol/L), p = 0.003). ROC curves indicated that plasma ACTH < or = 7.55 pmol/L distinguished patients in remission from treatment failures with 80% sensitivity and 97.4% specificity, and serum cortisol < or = 585 nmol/L with 100% sensitivity and 90% specificity. CONCLUSIONS: Twenty-four hours after transsesphenoidal surgery for Cushing's disease, and without glucocorticoids replacement, patients with serum cortisol concentrations higher than 585 nmol/L, and/or plasma ACTH higher than 7.55 pmol/L, and/or those in which an adenoma is not identified in the histological study, have a high risk of treatment failure. PMID- 17406781 TI - Surgical treatment of symptomatic cavernous malformations of the brainstem. AB - INTRODUCTION AND OBJECTIVES: Cavernous malformations (CM) at the level of the brainstem, continue to present a challenge in therapeutic terms and are an important source of controversy. Here we present our experience and the results obtained by adopting surgical treatment. MATERIALS AND METHODS: The results of a consecutive series of 17 patients were studied. The surgical intervention was designed after: 1. A neurological examination. 2. MRI and cerebral angiography. 3. Correlation with anatomical brainstem maps. The surgical intervention was approached from the most damaged zone or through a zone which was functionally least important. RESULTS: Complete extirpation was achieved in 15 patients without mortality. In a few patients the surgical intervention temporarily aggravated the prior lesion of the cranial nerves (2/17) or damage new sensory tracts (2/17). The functional post-operative recovery was good, in terms of consciousness (4/5), cranial nerves (11/17), the pyramidal tract (3/5) and the cerebellum (2/4). Of the patients that were operated, 14 of 17 returned to their professional activities. CONCLUSIONS: The results of surgery can surpass the morbidity-mortality of the natural history or treatment with radiosurgery. There is a clear consensus in recommending surgical intervention for CMs that are superficially located, in young patients and in those with a risk of further bleeding. It is probably best that the surgery is performed during the subacute period, when the MRI offers a clear image confirming the presence of the CM. PMID- 17406782 TI - Kinetics of salt-dependent unfolding of [2Fe-2S] ferredoxin of Halobacterium salinarum. AB - The [2Fe-2S] ferredoxin from the extreme haloarchaeon Halobacterium salinarum is stable in high (>1.5 M) salt concentration. At low salt concentration the protein exhibits partial unfolding. The kinetics of unfolding was studied in low salt and in presence of urea in order to investigate the role of salt ions on the stability of the protein. The urea dependent unfolding, monitored by fluorescence of the tryptophan residues and circular dichroism, suggests that the native protein is stable at neutral pH, is destabilized in both acidic and alkaline environment, and involves the formation of kinetic intermediate(s). In contrast, the unfolding kinetics in low salt exhibits enhanced rate of unfolding with increase in pH value and is a two state process without the formation of intermediate. The unfolding at neutral pH is salt concentration dependent and occurs in two stages. The first stage, involves an initial fast phase (indicative of the formation of a hydrophobic collapsed state) followed by a relatively slow phase, and is dependent on the type of cation and anion. The second stage is considerably slower, proceeds with an increase in fluorescence intensity and is largely independent of the nature of salt. Our results thus show that the native form of the haloarchaeal ferredoxin (in high salt concentration) unfolds in low salt concentration through an apparently hydrophobic collapsed form, which leads to a kinetic intermediate. This intermediate then unfolds further to the low salt form of the protein. PMID- 17406783 TI - Construction of a high-density and high-resolution human chromosome X array for comparative genomic hybridization analysis. AB - The human chromosome X is closely associated with congenital disorders and mental retardation (MR), because it contains a significantly higher number of genes than estimated from the proportion in the human genome. We constructed a high-density and high-resolution human chromosome X array (X-tiling array) for comparative genomic hybridization (CGH). The array contains a total of 1,001 bacterial artificial chromosome (BACs) throughout chromosome X except pseudoautosomal regions and two BACs specific for Y. In four hybridizations using DNA samples from healthy males, the ratio of each spotted DNA was scattered between -3SD and 3SD, corresponding to a log(2) ratio of -0.35 and 0.35, respectively. Using DNA samples from patients with known congenital disorders, our X-tiling array was proven to discriminate one-copy losses and gains together with their physical sizes, and also to estimate the percentage of a mosaicism in a patient with mos 45,X[13]/46,X,r(X)[7]. Furthermore, array-CGH in a patient with atypical Schinzel Giedion syndrome disclosed a 1.1-Mb duplication at Xq22.3 including a part of the IL1RAPL2 gene as a likely causative aberration. The results indicate our in-house X-tiling array to be useful for the identification of cryptic copy-number aberrations containing novel genes responsible for diseases such as congenital disorders and X-linked MR. PMID- 17406784 TI - Outcome of day-case inguinal hernia in elderly patients: how safe is it? AB - INTRODUCTION: Inguinal hernia repair is well established as a day-case procedure but little is reported about the outcome for elderly patients. METHODS: A retrospective study was undertaken to compare a study group of patients aged 70 and above with a controlled group of patients aged less than 65 who had day-case inguinal hernia repair (Lichtenstein repair) between January 1996 and December 2004. RESULTS: A total of 588 patients (282 in the elderly group and 306 in the control group) were studied. Although there were a larger number of higher-risk (ASA III) patients in the elderly group (P < 0.001), unplanned admission, postoperative symptoms, and satisfaction were comparable with those for the younger group. CONCLUSION: Day-case inguinal hernia repair can be safely offered to patients aged 70 and above. PMID- 17406785 TI - Inguinal hernia treatment with the Prolene Hernia System in a Dutch regional training hospital. AB - INTRODUCTION: In this retrospective study results from inguinal hernia repair with the Prolene Hernia System (PHS) in a regional training hospital were analysed. PATIENTS AND METHODS: One-hundred and seventy-eight primary inguinal hernias and thirty-nine recurrent hernias (initial non-mesh repair) were treated with the PHS. The primary endpoint was the recurrence rate. Secondary endpoints were short-term and long-term complications. Pain was evaluated by use of a visual analog scale (VAS, 0-100), and a short-form 36-item questionnaire was used to assess postoperation quality of life. All patients visited the outpatient clinic for a physical examination (100% follow-up). RESULTS: After a median follow-up of 32 months four patients were diagnosed with recurrent herniation (1.8%), three after primary hernia repair (1.6%) and one after recurrent hernia repair (2.6%). Three superficial wound infections (1.3%), three haematomas needing surgical evacuation (1.3%), and two lesions of the spermatic cord (0.9%) were diagnosed. Seven patients (3.2%) suffered from persistent pain (VAS > 40). Average VAS score was 13 (0-80) >24 months after surgery. CONCLUSION: In a regional training hospital, primary and recurrent inguinal hernias were treated with low recurrence and few complications by use of the PHS. PMID- 17406786 TI - Incarcerated Spigelian hernia mimicking obstructing colon carcinoma. AB - Spigelian hernia is a ventral abdominal hernia that only rarely causes incarceration or strangulation of the bowel. There are few reports in the surgical literature of colonic obstruction secondary to incarcerated Spigelian hernia. In this paper, we present a patient with an incarcerated sigmoid colon in a Spigelian hernia sac, mimicking on contrast enema an obstructing carcinoma. Accurate diagnosis was made pre-operatively by computed tomography (CT), and the hernia was repaired by polypropylene mesh in a tension-free manner. PMID- 17406787 TI - Inguinal hernia is a common complication in lower midline incision surgery. AB - Inguinal hernia is a known complication after radical retropubic prostatectomy (RRP). We have investigated whether other types of lower midline incision surgery in males increase the risk of inguinal hernia. Male patients operated with open prostatectomy for benign prostate hyperplasia (n = 95), pelvic lymph node dissection for staging of prostate cancer (n = 88), or cystectomy for bladder cancer (n = 76) were identified and were sent questionnaires in which they were asked about postoperative inguinal hernia morbidity. Two-hundred and seventy-one men operated with RRP had previously received a similar questionnaire. The answers were compared with those from a control group of 953 men who had not undergone surgery. Annual attributional hernia morbidity and Kaplan-Meier hernia free survival were calculated. The cumulative incidence of post-operative inguinal hernia and annual attributional hernia morbidity after the respective surgical procedures were clearly higher during the early years post-operation than for nonoperated patients. Inguinal hernia is a common postoperative complication in males after all the lower midline incision surgery investigated. PMID- 17406788 TI - Angiogenesis in primary central nervous system lymphoma (PCNSL). AB - Angiogenesis and angiogenic growth factors have a major role in the pathogenesis of malignancies. However, very little is known about the clinical and histopathological relevance of angiogenesis in primary central nervous system lymphoma (PCNSL). We investigated that expression of vascular endothelial growth factor (VEGF) of the lymphoma cells and microvessel density (MVD) were examined in 19 patients with PCNSL. Additionally, the presence of the blood-brain barrier (BBB) was examined using immunohistochemistry and the electron microscopy. MVD was significantly higher in nine cases with VEGF immunoreactivity (VEGF+) than in ten cases with negative immunoreactivity for VEGF (VEGF-) (P < 0.001). VEGF expression was significantly associated with a longer survival (P < 0.005). BBB markers were negative in angiogenic vessels of VEGF+. BBB markers were identified in vessels surrounding tumor cells and tight junctions were also preserved in the capillary endothelium surrounding tumor cells in VEGF-. Angiogenesis is associated with VEGF expression and an absent BBB in the vessels of PCNSL. The BBB may be preserved in lesions with lymphoma cell infiltration, especially in VEGF- PCNSL. VEGF may have a prognostic effect in PCNSL. PMID- 17406789 TI - Gliosarcoma with primitive neuroectodermal differentiation: case report and review of the literature. AB - The clinical and pathological features of a gliosarcoma with a primitive neuroectodermal component in a 52-year-old male are presented. To our knowledge, only three other cases of such an entity have been reported in the English literature. PMID- 17406790 TI - Choroid plexus acinar adenoma: a case report. AB - Mucus-secreting adenomas or acinar adenoma of the choroid plexus are very rare. We report the case of a 79-year-old male with a 3-year history of occipital headaches with vomiting, ataxia and cerebellar signs. He was first seen due to difficulty while walking. He was admitted to the hospital with significant tumor expansion and clinical deterioration. CT and MRI revealed obstructive hydrocephalus secondary to a large fourth ventricular cyst mass, which enhanced markedly on contrast administration. Pathological findings were consistent with an acinar choroid plexus adenoma. The tumor was attached to the ependymal lining and was strongly adhered to the walls and floor of the IV ventricle. Post operative bleeding complicated partial removal of this tumor. The patient died 6 h after surgery. PMID- 17406791 TI - Mitochondrial acyl carrier proteins in Arabidopsis thaliana are predominantly soluble matrix proteins and none can be confirmed as subunits of respiratory Complex I. AB - Arabidopsis mitochondria are predicted to contain three acyl carrier proteins (ACPs). These small proteins are involved in fatty acid and lipoic acid synthesis in other organisms and have been previously reported to be subunits of respiratory Complex I in mitochondria in mammals, fungi and plants. Recently, the mammalian mitochondrial ACP (mtACP) has been shown to be largely a soluble matrix protein but also to be minimally associated with Complex I (Cronan et al. 2005), consistent with its involvement in synthesis of lipoic acid for TCA cycle decarboxylating dehydrogenases in the matrix but contrary to earlier claims it was primarily a Complex I subunit. We have investigated the localization of the ACPs in Arabidopsis mitochondria. Evidence is presented that mtACP1 and mtACP2 dominate the ACP composition in Arabidopsis mitochondria, and both are present in the mitochondrial matrix rather than in the membrane. No significant amounts of mtACPs were detected in Complex I isolated by blue native gel electrophoresis, rather mtACPs were detected at low molecular mass in the soluble fraction, showing that in A. thaliana mtACPs are predominately free soluble matrix proteins. PMID- 17406792 TI - Transcriptional regulation of genes involved in the pathways of biosynthesis and supply of methyl units in response to powdery mildew attack and abiotic stresses in wheat. AB - From a library of 3,000 expression sequence tags (ESTs), derived from the epidermis of a diploid wheat (Triticum monococcum) inoculated with Blumeria graminis f. sp. tritici (Bgt), we cloned 23 cDNAs representing 12 genes that are involved in the pathways of biosynthesis and supply of methyl units. We studied the transcription of these genes to investigate how the methyl units are generated and regulated in response to Bgt infection and abiotic stresses in wheat. Expression of 5, 10-methylene-tetrahydrofolate reductase, methionine synthase, S-adenosylmethionine synthetase, and S-adenosylhomocystein hydrolase transcripts were highly induced at an early stage of infection. This induction was specific to the epidermis and linked to host cell wall apposition (CWA) formation, suggesting that the pathways for generation of methyl units are transcriptionally activated for the host defense response. Levels of S adenosylmethionine decarboxylase, caffeic acid 3-O-methyltransferase, 1 aminocyclopropane-1-carboxylate oxidase mRNA, but not phosphoethanolamine N methyltransferase and nicotianamine synthase mRNA, were up-regulated after infection and showed similar expression patterns to genes involved in the pathways of generation of methyl units, revealing possible routes of methyl transfer towards polyamine, lignin and ethylene biosynthesis rather than glycine betaine and nicotianamine in response to Bgt attack. After imposing various abiotic stresses, genes involved in the pathways of generation and supply of methyl units were also up-regulated differentially, suggesting that the generation of sufficient methyl units at an early stage might be crucial to the mitigation of multiple stresses. PMID- 17406793 TI - Identification of the ADP-glucose pyrophosphorylase isoforms essential for starch synthesis in the leaf and seed endosperm of rice (Oryza sativa L.). AB - ADP-glucose pyrophosphorylase (AGP) catalyzes the first committed step of starch biosynthesis in higher plants. To identify AGP isoforms essential for this biosynthetic process in sink and source tissues of rice plants, we analyzed the rice AGP gene family which consists of two genes, OsAGPS1 and OsAGPS2, encoding small subunits (SSU) and four genes, OsAGPL1, OsAGPL2, OsAGPL3 and OsAGPL4, encoding large subunits (LSU) of this enzyme heterotetrameric complex. Subcellular localization studies using green fluorescent protein (GFP) fusion constructs indicate that OsAGPS2a, the product of the leaf-preferential transcript of OsAGPS2, and OsAGPS1, OsAGPL1, OsAGPL3, and OsAGPL4 are plastid targeted isoforms. In contrast, two isoforms, SSU OsAGPS2b which is a product of a seed-specific transcript of OsAGPS2, and LSU OsAGPL2, are localized in the cytosol. Analysis of osagps2 and osagpl2 mutants revealed that a lesion of one of the two cytosolic isoforms, OsAGPL2 and OsAGPS2b, causes a shrunken endosperm due to a remarkable reduction in starch synthesis. In leaves, however, only the osagps2 mutant appears to severely reduce the transitory starch content. Interestingly, the osagps2 mutant was indistinguishable from wild type during vegetative plant growth. Western blot analysis of the osagp mutants and wild type plants demonstrated that OsAGPS2a is an SSU isoform mainly present in leaves, and that OsAGPS2b and OsAGPL2 are the major SSU and LSU isoforms, respectively, in the endosperm. Finally, we propose a spatiotemporal complex model of OsAGP SSU and LSU isoforms in leaves and in developing endosperm of rice plants. PMID- 17406794 TI - Cloning and characterization of a ribosomal protein L23a from Haemaphysalis qinghaiensis eggs by immuno screening of a cDNA expression library. AB - A primary cDNA library with a size of 1.34 x 10(6) PFU was constructed from Haemaphysalis qinghaiensis eggs and was immunoscreened with rabbit anti-H. qinghaiensis serum. One clone (Hq22, named following those clones obtained from adult Haemaphysalis qinghaiensis cDNA library which we constructed before) screened from the cDNA library was selected randomly for sequencing. The entire sequence of the clone was subsequently obtained using rapid amplification of the cDNA ends (RACE). A search of the cloned sequence against GenBank revealed that it related to ribosomal protein L23a (Rpl23a) and had a high percentage similarity to this protein from different species. Conserved domains for Rpl23a were also identified in the cloned sequence. Expression analysis by RT-PCR showed that this gene is expressed in salivary glands, midguts, other tissues and different developmental stages of H. qinghaiensis. Based on the H. qinghaiensis Rpl23a sequence, open reading frames (ORF) of Rpl23a of Heamaphysalis longicornis and Boophilus microplus were also cloned and were performed for comparison with Rpl23a of H. qinghaiensis and other organisms as well. Vaccine based on Rpl23a recombinant protein cannot protect sheep against H. qinghaiensis. PMID- 17406795 TI - The molecular basis of the Amblyomma americanum tick attachment phase. AB - Towards discovery of molecular signaling cascades that trigger and/or facilitate the tick attachment and formation of its feeding lesion, suppressive subtractive hybridization, high throughput sequencing and validation of differential expression by cDNA dot blot hybridization were performed on Amblyomma americanum ticks that had attained appetence and were exposed to feeding stimuli. This approach allowed for identification of 40 genes that are up regulated before ticks begin to penetrate the host skin. Based on BLAST and secondary structure homology searches as well as motif scan analyses, provisional identification was assigned to approximately 38% (15/40) of the identified genes that have been classified into 6 groups: Ligand binding (2 insulin-like growth-factor binding, lipocalin/histamine binding), immune responsive (tumor necrosis receptor associated factor 6, Microplusin-like antimicrobial), stress response proteins (Heat shock protein [HSP] 90, HSP40, 78 kDa glucose regulated protein [GRP78]), transporter polypeptides (ABC transporter and organic anion transporter polypeptide [contains Kazal-type serine proteinase inhibitor domain]) and enzymes/regulators (extracellular matrix metaloprotease inducer, chitinase), extracellular matrix-like proteins (tropoelastin, flagelliform silk protein). Sixty-two percent (25/40) of genes that did not show similarity to known proteins are classified as orphans. BLASTN homology search against the tick EST database revealed that 50% (20/40) of candidate genes are conserved in other ticks suggesting that molecular events underlying the A. americanum tick attachment phase may be conserved in other tick species. Consistent with the general assumption that genes that are up regulated in ticks before they started to penetrate host skin represented the tick's molecular preparedness to evade host defense during the attachment phase, real time RT-PCR analyses data demonstrated that the majority of the tested genes (9/11) were highly expressed during the first 24 h of feeding. Identification of genes in this study provides the framework for future studies to elucidate molecular signaling cascades that regulate early molecular events during the tick attachment phase. PMID- 17406796 TI - Significance of radiation therapy for adenocarcinomas of the esophagus, gastroesophageal junction and gastric cancer with special reference to the MAGIC trial. PMID- 17406797 TI - Austrian breast cancer patterns-of-care studies PCS93 and PCS01 versus PCS85 to identify changes in national practice. AB - PURPOSE: To determine the patterns of evaluation and treatment in Austrian breast cancer patients treated with surgery and radiotherapy in 1993 (PCS93) and 2001 (PCS01), and to compare these with the results of PCS85. MATERIAL AND METHODS: According to the evaluation process of the Austrian PCS85, ten randomly selected patient charts from every Austrian radiotherapy center from 1993 (n=100) as well as 2001 (n=120) were reviewed. The work-up included surgical and (histo)pathologic information, systemic therapy and detailed information on radiation therapy. RESULTS: Availability of histopathologic core data improved distinctly between 1985 and 2001. In 1985, treatment planning included fluoroscopic simulation and/or computed tomography in 55% of the cases as compared to 100% in 2001. The technical equipment in Austria changed distinctly. In 2001, 84% of the breast/chest wall fields and all regional node fields (+/- electrons) were treated by photons. Radiotherapy of the supraclavicular (77% vs. 28%), internal mammary (62% vs. 7.5%) and axillary nodes (51% vs. 5%) diminished from 1985 to 2001. A tumor bed boost was given in 34% of the patients in 1985 compared to 73% in 2001. The use of wedges (21% vs. 97%) and the number of port films (26% vs. 90%) increased substantially. CONCLUSION: Comparing both recent Austrian Patterns-of-Care surveys to the results obtained in 1985, an obvious increase in the quality level of histopathologic reporting as well as radiation treatment planning and delivery was found. PMID- 17406798 TI - Organ-sparing treatment of advanced bladder cancer: paclitaxel as a radiosensitizer. AB - BACKGROUND AND PURPOSE: Transurethral resection of bladder tumor (TUR-BT) and radiochemotherapy with cisplatin achieve high rates of bladder preservation and survival figures identical to radical cystectomy in muscle-invasive bladder cancers. The authors have investigated the potential use of paclitaxel in a radiochemotherapy protocol for patients with inoperable bladder carcinomas and mainly contraindications to cisplatin. PATIENTS AND METHODS: Between October 1997 to August 2004, 42 patients (median age 71 years) suffering from muscle-invasive (n=32) or recurrent (n=10) bladder cancers were treated with a paclitaxel containing radiochemotherapy (paclitaxel 25-35 mg/m(2) twice weekly) after TUR-BT (R0/1/2/x in n=18/4/14/3) or cystectomy with residual tumor (n=3). Five patients received additional cisplatin. Radiation treatment was administered to a total dose of 45-60 Gy. RESULTS: 76.2% completed the planned regimen. Adaptations of treatment were mainly required due to diarrhea. Grade 3/4 toxicities occurred in 15/1 patients. Severe renal toxicities did not occur. 28 patients underwent restaging TUR-BT 6 weeks after radiochemotherapy (complete remission/partial remission/progressive disease: n=24/3/1). Three patients developed a local recurrence and four distant metastases. Seven patients died from tumor, six of other reasons. CONCLUSION: Radiochemotherapy with paclitaxel was feasible and this bladder approach needs further investigation to evaluate whether paclitaxel could become a substitute for cisplatin. PMID- 17406799 TI - [Five year survival of patients with primary oral squamous cell carcinoma. Comparison of two treatment protocols in a prospective study]. AB - BACKGROUND AND PURPOSE: In recent years, different concepts for the treatment of oral squamous cell carcinomas (OSCC) have been developed; these include preoperative simultaneous neoadjuvant radiochemotherapy and one-stage surgery with tumor ablation and reconstruction. When considering long-term survival, there is substantial evidence that multimodality treatment based on a neoadjuvant radiochemotherapy is superior to adjuvant therapy concepts based on a surgical approach with postoperative irradiation. The aim of this study was to discuss the 5-year survival rate in a neoadjuvant and an adjuvant combination treatment in patients with primary OSCC. PATIENTS AND METHODS: This nonrandomized longitudinal study prospectively evaluates the long-term tumor-free survival in 128 patients with oral cancer. Two groups consisting of 74 neoadjuvantly and 54 primarily surgically treated patients were formed. 99 patients suffered from stage III and IV disease according to the UICC criteria. Long-term survival was estimated according to the Kaplan-Meier assumption. RESULTS: The neoadjuvant treatment increases the prospect of a long-term tumor-free survival. According to Kaplan Meier assumption the estimation for a 5-year tumor-free survival in OSCC in category T1 is 83.1% in neoadjuvant, and 70.1% in adjuvant treatment, in T2 79.6% and 57.7%, in T3 68.2% and 33.2%, in T4 51.4% and 30.5%, respectively. Significance (p<0.05) could be proven for T1 (p=0.002), T2 (p=0.028), and T4 (p<0.0001) tumors. The effectiveness of the preoperative radiochemotherapy was demonstrated in the pathohistological result of tumor-free resection specimens in 28 patients of the neoadjuvant treatment group (37.8%). On the other hand, four patients died during the preoperative combination therapy. 64.8% of the patients in the adjuvant and 71.6% in the neoadjuvant treatment group survived the observation period. CONCLUSION: Neoadjuvant therapy is highly effective and results in a better 5-year survival rate than adjuvant treatment. All patients with primary OSCC should be selected for such a treatment. Future therapy concepts should combine the effectiveness of a systemic treatment with the safety of a limited surgical resection of the tumor site to preserve as much function as possible. PMID- 17406800 TI - [Relevance of bisphosphonate long-term therapy in radiation therapy of endosteal jaw metastases]. AB - BACKGROUND AND PURPOSE: Bisphosphonates are used in the treatment of bone metastasis and malignant myeloma. They have been associated with osteonecrosis of the jaw (ONJ) since the year 2003. Etiology and pathogenesis of this clinical problem are not clear. The high rate of newly diagnosed cases is alarming. Based on the collective of two departments of oral and maxillofacial surgery, the results of therapy in the coincidence of systemic bisphosphonate treatment and local radiotherapy (RT) are analyzed. PATIENTS AND METHODS: In the course of 33 months (01/2003-09/2005), 63 synchronic or metachronic ONJs were seen in 42 patients as newly diagnosed episodes. Three of the 42 patients had an RT of jaw metastasis. RESULTS: Only one patient was followed over the whole treatment period. Under RT she developed a therapy-refractory ONJ similar to osteoradionecrosis at a dose of 40 Gy. She required a wide segmental osteotomy of the mandible resulting in a total loss of masticatory function. In 39 patients with 60 ONJs without RT not a single segmental resection of the mandible was necessary. CONCLUSION: Local RT in bisphosphonate long-term therapy seems to be a high risk constellation. As long as the pathogenesis of bisphosphonate-associated ONJ is unclear, patients subjected to RT of the head-and-neck area should be examined and dental findings should be treated. PMID- 17406801 TI - Radiosensitization dependent on p53 function in bronchial carcinoma cells by the isoflavone genistein and estradiol in vitro. AB - BACKGROUND AND PURPOSE: Simultaneous radiotherapy with chemotherapy is a standard treatment for inoperable non-small cell lung cancer (NSCLC), but the clinical outcome still remains poor. To further intensify treatment, substances need to be identified, which increase the effect of radiation on tumor cells without further enhancing toxicity to normal tissue. Hormones have a different toxicity profile than radiation or cytostatic drugs. As NSCLC often express estrogen receptors (ERs), the combination of genistein or estradiol and radiation in vitro was investigated. MATERIAL AND METHODS: A549 NSCLC cells with an inducible expression of a mutated TP53 and fibroblasts of a male donor (DF-18) were examined. ER expression was immunocytologically confirmed in all studied cell lines. Clonogenic survival was measured after incubation of the cells with genistein or estradiol (0.01 microM and 10 microM as maximum clinically applicable dose) and irradiation with different doses (0-4 Gy). The differentiation state of fibroblasts after combined therapy was analyzed. RESULTS: A549 cells expressing mutated TP53 were more radioresistant than TP53 wild-type cells. Incubation of nonfunctional TP53 cells with genistein or estradiol increased radiosensitivity in both tested concentrations. By contrast, radiosensitivity of A549 with wild type TP53 and DF-18 was not altered by hormonal incubation. In DF-18 radiation induced growth arrest that was not increased by additional hormonal incubation. CONCLUSION: NSCLC cells with nonfunctional TP53 might be sensitized against radiation by genistein or estradiol. As genistein is better tolerable than estradiol in patients, additional studies are warranted to assess potential gains of this combination therapy. PMID- 17406802 TI - [Dosimetric impact of image-guided translational isocenter correction for 3-D conformal radiotherapy of the prostate]. AB - BACKGROUND AND PURPOSE: Interfractional prostate motion during radiotherapy due to variation in rectal distension can have negative consequences. The authors investigated the dosimetric consequences of a linear translational position correction based on image guidance when a three-dimensional conformal treatment technique was used. MATERIAL AND METHODS: Planning CTs of seven patients with empty and distended rectum were analyzed. A reference plan for the planning target volume (PTV) and the boost were calculated on the CT dataset with the empty rectum with a standard four-field technique. The treatment plan was transferred to the CT with the distended rectum for an uncorrected setup (referenced to bony anatomy) and a corrected setup after position correction of the isocenter. The dosimetric consequences were analyzed. RESULTS: Organ motion decreased the coverage of the prostate by the 95% isodose during simulated single treatment fractions by up to -21.0 percentage points (%-p; boost plan) and by up to -14.9%-p for the seminal vesicles (PTV plan). The mean rectum dose increased by up to 18.3%-p (PTV plan). Linear translational correction (mean 6.4 +/- 3.4 mm, maximum 10.8 mm) increased the coverage of the prostate by the 95% isodose by up to 12.7%-p (boost plan), while the mean rectum dose was reduced by up to -8.9% p (PTV plan). For the complete treatment a reduction of complication probability of the rectum of approximately 5%-p was calculated. CONCLUSION: The use of an image guidance system with linear translational correction can improve radiation treatment accuracy for prostate cancer, if geometric changes are within certain limits. PMID- 17406803 TI - Helmet mold-based surface brachytherapy for homogeneous scalp treatment: a case report. AB - BACKGROUND/CASE REPORT: External-beam radiotherapy of complex-shaped areas is sometimes difficult to realize. In a patient with chronic lymphatic leukemia (CLL) infiltrates of the skin of the whole scalp, conventional external-beam radiotherapy with electrons or photons was not able to treat the target sufficiently. Thus, the authors developed a brachytherapy moulage technique using a customized helmet (polyethylene) with flexible interstitial plastic applicators and irradiated the target very homogeneously with a microselectron (192)Ir HDR (high-dose-rate) source (2.0 Gy daily fractionated; 36.0 Gy total dose related to the reference points in 3 mm focus depth). Technical difficulties, CT-supported three-dimensional conformal treatment planning, and verification with TLD probe measurements are described. The treatment was well tolerated and resulted in complete local remission of the CLL infiltrates within a follow-up of 30 months. CONCLUSION: The presented treatment of lymphoma infiltrates at the scalp by HDR moulage techniques is exceptional but safe and practicable to achieve local tumor control. PMID- 17406804 TI - Radiation recall pneumonitis induced by gemcitabine. AB - BACKGROUND: Radiation recall pneumonitis describes a very rare reaction in a previously irradiated area of pulmonary tissue after application of pharmacological agents. A case of recall pneumonitis induced by gemcitabine is reported. CASE REPORT: A 64-year-old female patient with metastasized esophageal carcinoma received simultaneous chemoradiotherapy of the upper mediastinum with 50.4 Gy and cisplatin/5-fluorouracil. 8 months later she was scheduled for salvage chemotherapy with gemcitabine (1,000 mg/m(2) days 1 and 8) and docetaxel (75 mg/m(2) day 8) due to locally progressive disease. After having received gemcitabine on day 1 of the second course, the patient developed dry cough, subfebrile temperatures and dyspnea within 48 h. A CT of the thorax revealed newly developed bilateral pulmonary ground-glass opacity corresponding to the previous radiation fields. Chemotherapy was stopped and systemic application of prednisolone was initiated. 2 months later symptoms had resolved with a control CT of the thorax showing complete regression of the pulmonary changes. CONCLUSION: Gemcitabine-induced recall pneumonitis is a rarely reported phenomenon and should be taken into account even after extended time interval to the previous radiotherapy. PMID- 17406805 TI - Expression of Toll-like receptor 4 in the brain in a rabbit experimental subarachnoid haemorrhage model. AB - OBJECTIVE: To investigate the expression of the Toll-like receptor (TLR) 4 in the brain after experimental subarachnoid haemorrhage (SAH) in rabbits. METHODS: A total of 52 rabbits were randomly divided into four groups: control group; day 3, day 5, and day 7 groups. Day 3, day 5, and day 7 groups were all SAH groups in which the rabbits were killed on day 3, 5, and 7, respectively. In SAH groups, autologous arterial blood was injected into cisterna magna twice on day 0 and day 2. Immunostaining and immunoblotting experiments were performed to detect the expression of TLR4 protein. Reverse-transcriptase polymerase chain reaction was used to analyze the presence and quantity of TLR4 mRNA. RESULTS: The expressions of TLR4 protein and mRNA were increased remarkably in SAH groups compared with the control group. The immunohistochemical staining demonstrated high level expression of TLR4 was present mainly in the endothelial cells of capillaries in the brain. CONCLUSION: Our results indicate that TLR4 expression is upregulated in the brain after experimental SAH. PMID- 17406806 TI - Expression analysis of the TAB2 protein in adult mouse tissues. AB - BACKGROUND: The Interleukin-1 (IL-1) signaling component TAK1 binding protein 2 (TAB2) plays a role in activating the NFkappaB and JNK signaling pathways. Additionally, TAB2 functions in the nucleus as a repressor of NFkappaB-mediated gene regulation. OBJECTIVE: To obtain insight into the function of TAB2 in the adult mouse, we analyzed the in vivo TAB2 expression pattern. MATERIALS AND METHODS: Cell lines and adult mouse tissues were analyzed for TAB2 protein expression and localization. RESULTS: Immunohistochemical staining for TAB2 protein revealed expression in the vascular endothelium of most tissues, hematopoietic cells and brain cells. While TAB2 is localized in both the nucleus and the cytoplasm in cell lines, cytoplasmic localization predominates in hematopoietic tissues in vivo. CONCLUSIONS: The TAB2 expression pattern shows striking similarities with previously reported IL-1 receptor expression and NFkappaB activation patterns, suggesting that TAB2 in vivo is playing a role in these signaling pathways. PMID- 17406807 TI - The inhibitory effects of H+ K+ ATPase inhibitors on human neutrophils in vitro: restoration by a K+ ionophore. AB - OBJECTIVE: This study investigated the ability of proton pump inhibitors (PPI), such as omeprazole and pantoprazole, to inhibit neutrophil migration, calcium mobilization and the mechanisms involved in this inhibition. METHODS: Neutrophils were incubated with different concentrations of omeprazole and pantoprazole for 30 min and stimulated to migrate with fMLP and IL-8. Treatment toxicity was assessed by MTT assay. The intracellular calcium levels were analyzed in neutrophils pre-treated with omeprazole and pre-loaded with FURA-2AM, when stimulated with fMLP. The activity of p38 MAP Kinase was evaluated by Western blot after treatment with omeprazole. RESULTS: Omeprazole is able to inhibit neutrophil chemotaxis to fMLP and IL-8. Pantoprazole demonstrated the same ability. This inhibitory effect was not due to a toxic effect of the proton pump inhibitors. Inhibition of v-ATPase by bafilomycin did not modify the ability of fMLP or IL-8 to induce neutrophil migration. Omeprazole was also able to decrease intracellular calcium availability. The addition of a potassium ionophore, nigericin, restored the migratory ability, as well as the intracellular calcium levels. The activity of p38 MAP Kinase was decreased in neutrophils pretreated with omeprazole. CONCLUSION: Proton pump inhibitors promote inhibition of H+ K+ ATPase in neutrophils, resulting in cationic flow disturbances through the cellular membrane that, consequently, inhibit migratory and intracellular events such as calcium influx and p38 MAP Kinase activation. PMID- 17406808 TI - A simple preparation method for mouse eosinophils and their responses to anti allergic drugs. AB - OBJECTIVE AND DESIGN: A simple method for preparing mouse eosinophils was established, and the characteristics of the eosinophils were assessed including their responses to anti-allergic drugs. MATERIALS OR SUBJECTS: Mouse eosinophils were prepared from peritoneal exudate cells of BALB/c mice primed and boosted with antigen ovalbumin (OVA). METHODS: Surface phenotypes, migration activities and leukotriene C(4) (LTC(4)) production abilities of these eosinophils were examined. In addition, the effects of anti-allergic drugs, oxatomide and tranilast, on generation of LTC(4) from mouse eosinophils were examined. RESULTS: Eosinophils of mice boosted with OVA were phenotypically and functionally identical with human eosinophils. Around 1 x 10(7) eosinophils were obtained from mouse peritoneal exudate. It was found that these mouse eosinophils enabled to migrate in response to eotaxin as well as platelet-activating factor (PAF), and generated LTC(4) by IL-5 stimulation. Moreover, it was revealed that clinically used anti-allergic drugs inhibited LTC(4)-production dose-dependently. CONCLUSIONS: The present study provides a convenient method to obtain fully functional mouse eosinophils that are useful for drug screening and for evaluating implications of eosinophils in allergic responses. PMID- 17406809 TI - Oxidized fat induces oxidative stress but has no effect on NF-kappaB-mediated proinflammatory gene transcription in porcine intestinal epithelial cells. AB - OBJECTIVE AND DESIGN: The effect of oxidized frying oils on PPARgamma which is a potent inhibitor of inflammatory responses in the intestine is currently unknown. Thus, the present study aimed to explore the effect of oxidized frying oil on PPARgamma DNA-binding and proinflammatory responses in intestinal epithelial cells. MATERIAL AND METHODS: 18 male pigs were fed two different diets containing either fresh fat or oxidized fat (n = 9 each). After 28 d, intestinal epithelial cells were isolated and analyzed for PPARgamma DNA-binding, NF-kappaB DNA-binding and NF-kappaB target gene expression. In addition, markers of lipid peroxidation and antioxidant status were determined. RESULTS: Feeding the oxidized fat slightly, but not significantly, activated PPARgamma DNA-binding in intestinal epithelial cells when compared to fresh fat. In addition, oxidized fat increased the concentration of TBARS and reduced the concentrations of alpha-tocopherol and activities of antioxidant enzymes relative to fresh fat (P < 0.05). No effect of the oxidized fat was observed on NF-kappaB DNA-binding and NF-kappaB target gene expression in intestinal epithelial cells. CONCLUSIONS: The present study suggests that moderate PPARgamma activation and induction of oxidative stress by oxidized frying oil have no implication for NF-kappaB-mediated proinflammatory gene expression in porcine intestinal epithelial cells. PMID- 17406810 TI - Regulation of cytokine synthesis in cardiac surgery: Role of extracorporeal circuit and humoral mediators in vivo and in vitro. AB - OBJECTIVE AND DESIGN: Cardiopulmonary bypass (CPB) impairs monocyte and neutrophil proliferation, cytokine synthesis, and antigen presentation. This study compares in vivo data with results from an extracorporeal circulation (ECC) model, distinguishing direct effects on cytokine synthesis from regulatory mechanisms. PATIENTS AND METHODS: Whole blood from 18 patients prior to, during and after CPB was stimulated with lipopolysaccharide (LPS). Tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and IL-8 levels were measured. Additionally, blood from 4 volunteers was circulated in an ECC model. Cytokine levels were measured before and during mock ECC. RESULTS: LPS-induced cytokine synthesis was reduced after CPB (TNF-alpha: 11 %; IL-6: 29 %; IL-8: 48 % of preoperative values, all p < 0.001). In mock ECC, cytokine production (except IL-8) was suppressed: TNF-alpha production was lowest 60 min after starting ECC, IL-6 synthesis was lowest at 90 min (33 % and 15 % vs. pre-ECC levels; both p < 0.001). Patient sera contained cytokine-inhibitory activity after CPB, an activity not found in mock ECC. CONCLUSIONS: (1) In patients, CPB induces early transient LPS hyporesponsiveness; (2) blood contact with foreign surfaces induces LPS hyporesponsiveness; (3) serum cytokine-inhibitory activities are released after CPB, but not in mock ECC. Impaired leukocyte function may explain increased susceptibility to infections after CPB. PMID- 17406811 TI - [A comparison between absorbable and non-resorbable scleral implants in deep sclerectomy (T-Flux and SK-Gel)]. AB - BACKGROUND: The intraocular pressure (IOP) lowering effects of deep sclerectomy (partially combined with phacoemulsification) with different scleral implants (T Flux- or SK-Gel) were investigated. PATIENTS AND METHODS: In a retrospective study, 72 patients with medically uncontrollable glaucoma underwent non penetrating deep sclerectomy. Of these, 54 patients received T-Flux implants and 18 SK-Gel implants. Examinations were carried out shortly before and after surgery, as well as after 12 months. RESULTS: Prior to surgery IOP was 18.4+/-5.5 mmHg (n=72) and 12 months after surgery it was 13.1+/-3.8 mmHg (n=65). The number of antiglaucomatous eyedrops used prior to surgery was 2.3+/-1.3 (n=72) and 12 months after surgery 0.2+/-0.6 mmHg (n=65). Secondary IOP-lowering surgery after 12 months was carried out on 15.3% of the operated eyes, and consecutive goniopunctures after 12 months were 25%. No significant differences were found between the two groups. CONCLUSIONS: The short- and mid-term IOP lowering effects in deep sclerectomy with scleral implants were quite satisfying no matter which implant was used. There was no difference in deep sclerectomy whether or not combined with cataract surgery. PMID- 17406812 TI - [Rate of reopening of macular holes following cataract operation]. AB - BACKGROUND: We set up this retrospective study to examine how different the functional and morphological results are following pars plana vitrectomy (ppV) for idiopathic macular holes in phakic eyes with later cataract surgery and in pseudophakic eyes, and to what extent cataract surgery can induce reopening of idiopathic macular holes after ppV. METHODS: A total of 189 eyes were vitrectomized because of idiopathic macular hole (37 pseudophakic, 152 phakic); 120 (22 pseudophakic, 98 phakic) of these eyes were examined at follow-up of an average of 19 months after vitrectomy. Cataract surgery was performed in 65 of the phakic eyes in this period. The functional and morphological results recorded for these 22 pseudophakic and 65 phakic eyes were evaluated. RESULTS: In the pseudophakic eyes preoperative visual acuity was 0.14, increasing to 0.20 postoperatively (p=0.16); in 1 case (4.5%) the macular hole was not closed at the time of the follow-up examination. Cataract extraction was performed in 65 of the phakic eyes an average of 10 months after ppV. The initial VA was 0.19 before ppV and increased to 0.37 (p<0.01) after cataract surgery; in 4 cases (6.2%) the macular hole was not closed by the time of the follow-up examination, but in each case this had already been noted when the cataract extraction was done. Reopening did not occur after cataract extraction in any of these cases. CONCLUSION: With a rate of 5.7% for reopening or persistence of macular holes, our results are comparable to those recorded in other studies. Cataract surgery following ppV does not influence the reopening rate of macular holes. PMID- 17406813 TI - Safety and efficacy of aspartame-based liquid versus sucrose-based liquids used for dilution in oral sodium phosphate solutions for colonoscopy preparations. AB - The aim of this study was to investigate whether an oral sodium phosphate solution (OSPS) mixed with aspartame-based clear liquids as the diluent would yield improved colon cleansing results compared to an OSPS mixed with sucrose based liquids as the diluent. Fifty-one patients undergoing colonoscopy were prospectively randomized into two groups to receive different OSPS colonoscopy preparations, with sucrose-based or aspartame-based liquids used as diluents. The primary end point was the quality of the colonoscopy preparation and secondary end points were serum electrolytes before and after preparations. No significant difference in colonoscopy preparation quality was seen between the two OSPS diluent groups (Mantel-Haenzel chi (2) = 0.795, P = 0.484). There were no significant differences in mean electrolyte shifts of sodium, potassium, blood urea nitrogen (BUN), creatinine (Cr), or BUN/Cr ratios between the two groups. There was a statistically significant increase in serum phosphorous in the aspartame-based group compared to the sucrose-based diluent group (P = 0.021). In conclusion, there was no clinically detectable difference in colonoscopy preparation quality between the two OSPS diluent groups. This study suggests that passive fluid transport by aquaporins may well be the major mediator of fluid shifts in the study subjects. This result suggests the potential importance of aquaporins and minimizes the importance of sodium glucose cotransporter SGLT1 in fluid and electrolyte transport in the human gastrointestinal tract. Aspartame or its constituent amino acids may enhance phosphate absorption across the human small intestine. PMID- 17406814 TI - Age and indication for referral to capsule endoscopy significantly affect small bowel transit times: the given database. AB - The purpose of this study was to examine the effect of age and selected indications for capsule endoscopy on small bowel transit times. Data on 67 clinical studies (790 subjects with different gastrointestinal pathologies [49.5% males; mean age, 51.9 +/- 18.33 years; range, 18-91 years] and 87 healthy volunteers) were retrieved from the company (Given Imaging, Ltd.)-sponsored database. All subjects swallowed the PillCam SB Capsule after a 12-hr fast. The capsule reached the cecum in all 877 participants. Indications for referral for capsule endoscopy were as follows: 372 obscure gastrointestinal bleeding, 96 suspected Crohn's disease, 65 celiac disease, 54 irritable bowel syndrome, and 116 familial adenomatous polyposis, intestinal lymphoma, or ulcerative colitis. One group consisted of patients <40 years old (n = 235), and the other patients 40 years old (n = 555). The younger group, volunteers, and Crohn's disease patients had significantly shorter small bowel transit times than the others (P < 0.001). Gastric emptying indirectly influenced capsule transit time. PMID- 17406815 TI - Biliary tract obstruction secondary to malignant lymphoma: experience at a referral center. AB - Lymphoma is a rare cause of biliary obstruction and, on cholangiography, may mimic other causes of obstructive jaundice. The optimum treatment for these patients is unclear. The aim of this study is to evaluate the incidence, clinical and imaging findings, management, and outcome of biliary obstruction caused by lymphoma. Our database was searched retrospectively for patients with biliary obstruction due to lymphoma between 1999 and 2005. Biliary obstruction secondary to lymphoma was found in 7 (0.6%) of 1123 patients with malignant biliary obstruction. One patient had benign biliary obstruction related to lymphoma. Of the eight patients (five male, three female; mean age, 34.50 +/- 17.93 years), four had Hodgkin's disease and four had non-Hodgkin's lymphoma. Biliary obstruction occurred as part of the initial or early presentation of lymphoma in two patients. The most common cause of obstruction was compression of the biliary tract by enlarged lymph nodes (six patients). Cholangiographic appearances were diverse: narrowing of the common bile duct (six patients), splayed and narrowed common bile duct (one patient), and multiple strictures and dilatations of the intrahepatic bile ducts (one patient). Biliary drainage was performed in all patients including endoscopic stent placement in six patients, nasobiliary drainage in one, and choledochoduodenostomy in one. Hyperbilirubinemia resolved in all but one of the patients with a stent; however, none could be maintained in a stent-free condition. Five patients died within 1 year after onset of jaundice. One of the surviving patients developed a late benign stricture at the site of the earlier lymphoma. We conclude that lymphoma should be considered in the differential diagnosis of obstructive jaundice, particularly in younger patients. We suggest that biliary drainage by the endoscopic or percutaneous route is necessary for the treatment of these patients. Late benign strictures may develop. Biliary obstruction is a sign of poor prognosis in lymphoma. PMID- 17406816 TI - Cyclo-oxygenase-independent inhibition of apoptosis and stimulation of proliferation by leptin in human colon cancer cells. AB - Obesity increases the risk of colon cancer. Hyperleptinemia is characteristic of obesity and leptin has been reported to be a colonic growth factor. We have examined the involvement of the cyclo-oxygenase (COX) pathways in the proliferation and anti-apoptotic effects of leptin. Leptin stimulated proliferation in HT-29 colon cancer cells: this was unaffected by inhibition of COX-1, COX-2, protein kinase C, or the epidermal growth factor receptor. Leptin did not increase COX-2 mRNA or COX-derived prostaglandin E2 production. Celecoxib induced apoptosis in a COX-independent manner. Leptin reduced both serum starvation- and celecoxib-induced apoptosis. Inhibition of ERK, p38 MAP kinase, and nuclear factor (NF)-kappaB abolished the growth-promoting and anti-apoptotic effects of leptin. Treatment of HT-29 cells with leptin stimulated phosphorylation of ERK and p38 MAP kinase and nuclear translocation of active NF kappaB. We conclude that leptin stimulates colon cancer proliferation via COX independent pathways and reduces celecoxib-induced apoptosis via ERK, p38 MAP kinase, and NF-kappaB pathways. PMID- 17406817 TI - A novel Von Hippel-Lindau case with germline mutation at codon 167 (CGG to TGG) having endocrine microadenomatosis of the pancreas. PMID- 17406818 TI - Epstein-Barr virus-positive inflammatory pseudotumor of the spleen with concomitant rectal cancer: a case report and review of the literature. PMID- 17406819 TI - A case of diarrhea, ataxia, and capsule endoscope retention. PMID- 17406820 TI - Clostridium difficile colitis in solid organ transplantation--a single-center experience. AB - Clostridium difficile (CD) is one of the most common causes of diarrhea in solid organ transplantation (SOT). Between 1996 and 2005, a total of 2474 solid organ transplants were performed at our institution, of which 43 patients developed CD associated diarrhea. There were 3 lung, 3 heart, 20 liver, 8 kidney-pancreas, 6 kidney, 1 composite tissue, and 2 multivisceral recipients. Onset of CD infection ranged from 5 to 2453 days posttransplant. All patients presented with abdominal pain and watery diarrhea. Toxins A and B were detected using rapid immunoassay or enzyme immunoassay. Treatment consisted of reduction of immunosuppression, fluid and electrolyte replacement, metronidazole (n=20), oral vancomycin (n=20), and a combination of metronidazole and vancomycin (n=2). Toxic megacolon was seen in five patients. Two of them had colonoscopic decompression, and the remaining three required colonic resection. One of these patients died due to multiorgan failure after cured CD enteritis. The remaining patients were discharged with well-functioning grafts and all are currently alive. CD colitis was a rare complication prior to 2000; 38 of the 43 cases occurred thereafter. We conclude that CD colitis represents a severe complication following SOT. Recently, a dramatic increase in the incidence of this complication has been observed. The development of life-threatening toxic megacolon must be considered in solid organ recipients. PMID- 17406821 TI - Unrecognized GERD symptoms are associated with excessive daytime sleepiness in patients undergoing sleep studies. AB - Sleep disturbances are commonly reported by patients who suffer from gastroesophageal reflux disease (GERD) but it is uncertain if GERD plays a role in patients with sleep disorders of undetermined origin. The prevalence of GERD in patients with sleep disorders of unknown etiology is uncertain; the aim of this study was to determine this prevalence. Three hundred eighty-five consecutive patients reporting to an outpatient clinic for evaluation of sleep disorders were assessed for their sleepiness in relation to reflux symptom intensity. Reflux symptoms that met the survey criteria for a diagnosis of GERD were present in 45 of the 385 subjects (12.8%). These subjects did not have a diagnosis of GERD and were not being treated. Multiple regression analysis showed that excessive sleepiness was associated with intensity of GERD symptoms. Patients with GERD had significantly higher Epworth sleep scores than patients without GERD (12.8 vs. 10.6; p=0.007), indicating more daytime sleepiness. We conclude that unrecognized and untreated GERD are present in many patients presenting with sleep disorders. Patients with GERD had significantly greater sleepiness. Further studies of the impact of GERD treatment in this population are necessary. PMID- 17406822 TI - Antiviral response of HCV genotype 1 to consensus interferon and ribavirin versus pegylated interferon and ribavirin. AB - Achieving an antiviral response at a reasonable cost is a challenge in the treatment of patients with chronic hepatitis C. A previous study indicated that consensus interferon with ribavirin had promising activity against hepatitis C virus (HCV) genotype 1. The objective of this study was to determine the virologic response with consensus interferon or pegylated interferon alpha-2b plus weight-ribavirin in patients chronically infected with HCV genotype 1. Intention-to-treat analysis showed response in 37% and 41% of subjects treated with consensus interferon/ribavirin or pegylated interferon/ribavirin, respectively, with response rates of 42% and 44% observed in analysis of the per protocol population, not a significant difference. Tolerability of the two treatment regimens was similar. In conclusion, both treatment regimens were safe and gave a similar antiviral response. It is possible that if consensus interferon is administered daily rather than three times weekly, eradication of HCV could be achieved in a larger proportion of patients infected with HCV genotype 1. PMID- 17406823 TI - A systematic review of provider knowledge of hepatitis C: is it enough for a complex disease? AB - As studies indicate that patients with hepatitis C face poor provider knowledge and even stigmatization, we conducted a systematic review of provider knowledge about and attitudes toward hepatitis C. We searched Medline for original studies between 1990 and 2005. Articles were abstracted to define target population, recruitment strategies, study design, and key findings. Twenty-six publications performed in nine countries were identified. Whereas studies demonstrated an understanding of the nature of hepatitis C, significant knowledge deficits existed related to natural history, diagnostic approaches, and treatment. The relevance of simple measures, such as vaccinations against hepatitis A and B, was underappreciated. While providers were aware of risk factors for the disease, there were substantial misperceptions, with 5%-20% of providers considering casual contact as a risk for disease acquisition. We conclude that while healthcare providers understand the nature of hepatitis C, important knowledge gaps persist, which may constitute barriers to appropriate therapy. PMID- 17406824 TI - Sequential observations show upregulation of TGF-beta1 at the early phase of chronic small bowel rejection in rats. AB - Although TGF-beta1 is related to chronic rejection in various solid organs, its role remains controversial in the setting of small bowel transplantation. The aim of this study is to investigate the distinct role of TGF-beta1 and its receptors in the setting of chronic small bowel rejection. Sequential protocol biopsies were performed every 2 weeks from week 2 till week 8 after transplantation and recipients were sacrificed at week 16 as the end point. Gene expression was evaluated by SYBR Green I real-time polymerase chain reaction and protein was determined by immunohistochemistry. The gene expressions of TGF-beta1 and Type I receptor were upregulated and attained statistical significance at 4, 6 and 8 weeks in allografts. Dynamic expressions of TGF-beta1 and Type I receptor mRNA shown by sequential protocol biopsies support the view that TGF-beta1 signaling is involved in the pathogenesis of chronic small bowel rejection, especially at the early phase. PMID- 17406825 TI - Factors prognostic of survival in patients awaiting liver transplantation for end stage liver disease. AB - In this study, we investigated possible independent predictive factors for survival, other than MELD score, in patients with cirrhosis. We reviewed the serum sodium, cholesterol, albumin, and platelet levels of 99 patients with cirrhosis and investigated the possible correlation of these parameters with survival period. We found that 77% and 81% of patients with cirrhosis were hypocholesterolemic and hypoalbuminemic, respectively. We noted that the survival time of 6 months in patients with serum sodium levels <125 mM at the time of admission to the study was 27% less than that in patients with sodium levels >130 mM. Patients with cirrhosis and serum sodium levels >130 mM survived for more than 1 year (95% CI). MELD scores of patients with serum sodium levels >130, between 125 and 129, and <125 mM were 15.8 to 19.9, 19.7 to 23.6, and 23.3 to 27.2, respectively (95% CI). In conclusion, we suggest that although all of these parameters are correlated with survival in patients with cirrhosis, the serum sodium level is the most accurate prognostic factor and a valid tool for predicting survival when considered in combination with the MELD score. PMID- 17406826 TI - Prognostic significance of hepatitis C virus RNA detection by transcription mediated amplification with negative polymerase chain reaction during therapy with peginterferon alpha and ribavirin. AB - The lower limit of detection of most polymerase chain reaction (PCR) assays for hepatitis C virus (HCV) RNA is 50 IU/ml, compared to 5 IU/ml for the transcription-mediated amplification (TMA) method. We retrospectively reviewed 57 patients to assess the predictive value of a positive TMA in the setting of a negative PCR during antiviral therapy. Patients were divided into (1) PCR-/TMA+ (discordant; n=21) and (2) PCR-/TMA-(concordant; n=36). Sustained virologic response (SVR) was decreased in the discordant group (48% vs. 75%; P=0.04). In discordant patients, SVR was more frequent in patients who had one positive TMA than in those who had two or more positive TMAs or one positive TMA and recurrent HCV RNA detectability by PCR during treatment (78% vs. 25%; P=0.03). Breakthrough occurred more frequently in discordant patients (24% vs. 3%; P=0.02). A positive TMA on two or more occasions in patients who have become PCR-negative on therapy indicates a high likelihood of treatment failure. PMID- 17406827 TI - Increased age, male gender, and cirrhosis, but not steatosis or a positive viral serology, negatively impact the life expectancy of patients who undergo liver biopsy. AB - Most survival studies of chronic liver disease avoid including more than one condition, often present in patients with liver disease; survival analysis of patients with liver disease in general was undertaken. Over a 9-year period, the survival experience of 365 patients who underwent liver biopsy, with a median follow-up of 3 years, was assessed. Log rank tests and Cox regression were used to evaluate risk factors. The Flemington-Harrington G(rho) family of tests compared the number of deaths expected in the U.S. population in general, adjusted for age, sex, and year of biopsy, to the observed number of deaths in the patients with cirrhosis and to the observed number of deaths in patients without cirrhosis. Twenty-two (6%) patients died. Cirrhosis (HR = 2.9; 95% c.i.: 1.2-6.7), male sex (HR = 2.7; 95% c.i.: 1.1-6.6), and an additional 20 years of age at biopsy (HR = 2.9; 95% c.i:. 1.4-6.2) each negatively impacted survival. Patients with cirrhosis experienced 4.58 times the number of expected deaths (p < 0.00001). Patients without cirrhosis experienced 1.66 times the number of expected deaths (p = 0.15). Steatosis and a positive viral serology did not increase the risk of death. Male gender, increased age, and cirrhosis increased the risk of death; increased steatosis and positive viral serologic studies did not. PMID- 17406828 TI - Fatigue and health-related quality of life (HRQL) in chronic hepatitis C virus infection. AB - In addition to chronic hepatitis, many individuals infected with hepatitis C virus (HCV) suffer from fatigue, which may compromise their health-related quality of life (HRQL). To assess systematically health-related quality of life (HRQL) in patients with chronic hepatitis C and to determine if any clinical, biochemical, virologic, demographic, and histologic features are associated with HRQL status. In this cross-sectional observational study, one hundred thirty patients with chronic HCV infection (HCV RNA positive by PCR) and 61 healthy controls were enrolled from a tertiary care teaching medical center. All patients and controls completed one generic HRQL questionnaire (MOS SF-36) and one liver disease specific instrument (Chronic Liver Disease Questionnaire, CLDQ). Ninety five HCV patients and all the controls also completed a fatigue questionnaire (Chronic Fatigue Screener, CFS) and had immunologic markers determined (Cryoglobulin, Soluble IL-2 receptors, Rheumatoid Factor). We compared the HRQL of HCV-infected patients to the controls and, using data from other studies, to the general population, patients with diabetes, and patients with chronic low back pain. Patients with chronic HCV had greater HRQL impairment than healthy controls and those with type II diabetes. Fatigue was the most important symptom with negative impact on HRQL. Sixty-one percent of HCV-infected patients reported fatigue-related loss of activity. Additionally, other factors associated with HRQL were gender and histologic cirrhosis. Chronic HCV infection has a profound negative impact on patients' HRQL. Disabling fatigue is the most important factor that contributes to loss of well-being in this relatively young group of patients. PMID- 17406829 TI - Factors influencing treatment efficacy of 24-week combination therapy with interferon alpha-2b plus ribavirin for chronic hepatitis C. AB - The efficacy of a combination therapy of interferon (IFN) alpha-2b plus ribavirin (RBV) in chronic hepatitis C, and the factors contributing to efficacy, were investigated. One hundred eighty-six cases were enrolled in this study and treated with a combination of IFN alpha-2b plus RBV. IFN alpha-2b was administered at 6-10 mega-units daily for 2-4 weeks and three times per week for 20-22 weeks, in combination with oral intake of RBV at 600 or 800 mg for 24 weeks. Rates of sustained virological response (SVR) were 34.9% in serogroup 1 (SG1) and 82.5% in SG2. SVR rates in cases with both drugs discontinued, reduced, and unchanged were 4.2%, 40%, and 42.7% in SG1 and 42.9%, 76.9%, and 91.9%, in SG2. In terms of the total RBV dose per kilogram body weight, SVR rates were 14.3% and 46.2% with <1600 mg, 36.2% and 91.7% with 1600-2000 mg, and 62.1% and 95% with > or =2000 mg in SG1 and SG2, respectively. Total doses of RBV per body weight and negative HCV RNA at 8 weeks were the significant factors contributing to SVR in SG1 cases. These results indicate that it was critical to tailor the doses of RBV and IFN to the individual. To improve the rate of SVR, it is necessary to make efforts, where possible, not to discontinue drug use or reduce the drug dose. PMID- 17406830 TI - Gastroprotective effects of DA-6034, a new flavonoid derivative, in various gastric mucosal damage models. AB - This study evaluated the gastroprotective activity of DA-6034 against various ulcerogens including ethanol, aspirin, indomethacin, stress, and acetic acid. The basic mechanisms of DA-6034 as a defensive factor such as mucus secretion and endogenous prostaglandin E(2) synthesis were determined. Rats with gastric lesions induced by ethanol-HCl, aspirin, indomethacin, and stress that had been pretreated with DA-6034 orally showed a statistically significant decrease or decreasing tendency of the gastric lesion. In acetic acid-induced gastric lesions, repeated oral administration of DA-6034 exhibited a U-shape activity in ulcer healing, with the maximum and minimum inhibition being observed at 30 and 10 mg/kg/day, respectively. DA-6034 also increased the mucus content in the gel layer as well as endogenous prostaglandin E(2) synthesis. These results suggest that DA-6034 prevents gastric mucosal injury, and these gastroprotective activities appear to be due to the increase in the gastric defensive systems. PMID- 17406831 TI - The role of chromoendoscopy in the surveillance of the duodenum of patients with familial adenomatous polyposis. AB - Adenomas of the duodenum have been described in patients with familial adenomatous polyposis (FAP). Patients with FAP are at high risk for the development of periampullary cancer. The aim of our study was to evaluate if endoscopic visualization of small polyps, often overlooked at standard endoscopic examination, was improved by chromoendoscopy. Ten patients with FAP and previous colectomy underwent upper gastrointestinal endoscopy. Two skilled endoscopists were involved for each endoscopy. Evaluation of number and diameter of polyps was made before and after staining. After staining we detected a larger number of duodenal polyps than found at the standard endoscopic examination, the difference being statistically significant. This result seems to suggest that chromoendoscopy may improve diagnostic yield of endoscopy. Further studies are needed to suggest the best surveillance program and the appropriate therapeutic modality for these patients. PMID- 17406832 TI - Nodular gastritis in a patient with myopericarditis. PMID- 17406833 TI - Split-liver transplant for two adults in a spanish hospital. PMID- 17406834 TI - Serum total sialic acid in differential diagnostics of jaundice caused by malignant and nonmalignant diseases: a ROC curve analysis. AB - The purpose of this study was to evaluate the usefulness of serum total sialic acid (SA) for discrimination of malignant and nonmalignant jaundice. Serum SA concentration and its ratio with total protein (SA/TP) were determined in 55 patients with jaundice: 25 malignant and 30 nonmalignant. SA was estimated by enzymatic method. Serum total SA and the ratio SA/TP were significantly higher in malignant than in nonmalignant jaundice. Diagnostic sensitivity of SA and the ratio of SA/TP in both types of jaundice reached the value of 95.8%. The specificity, positive and negative predictive values, and efficiency of SA and SA/TP were higher in malignant than in nonmalignant jaundice. Areas under ROC curves for SA and the ratio of SA/TP in malignant jaundice were higher than in nonmalignant, but there were not statistically significant differences. SA levels and the ratio of SA/TP do not have the ability to discriminate between these types of jaundice. PMID- 17406835 TI - Effect of alcohol consumption on leptin level in serum, adipose tissue, and gastric mucosa. AB - It has been reported that the stomach is a source of leptin, which is the product of the obese (ob) gene. In the present study, the effect of alcohol on leptin level in serum, gastric mucosa, and adipose tissue was studied to understand the relationship between appetite and alcohol consumption. Male Sprague-Dawley rats were administered 1 ml of 25% ethanol perorally. Leptin levels in the serum, gastric mucosa, and adipose tissue were measured. The serum leptin level was significantly decreased 3 and 6 hr after ethanol administration, although the gastric leptin level was not affected. The leptin level in the adipose tissue was significantly increased 3 hr after administration. We conclude that the decreased serum leptin level after ethanol administration might be due to suppression of leptin secretion from adipose tissue to the systemic circulation. These findings might be important for understanding the relationship between alcohol consumption and appetite. PMID- 17406836 TI - Drinking capacity and severity of dyspeptic symptoms during a water load test after Nissen fundoplication. AB - Up to 30% of subjects undergoing Nissen fundoplication develop dyspeptic symptoms. Previous studies using the water load test have shown that functional dyspepsia patients have a diminished drinking capacity due to increased visceral sensitivity. Here we investigate drinking capacity and severity of symptoms in 9 patients with dyspeptic symptoms after fundoplication, 9 patients without symptoms after surgery, 18 functional dyspepsia patients, and 18 controls. The water load test was performed by having subjects drinking room-temperature tap water at a predetermined rate of 15 ml/min. The presence and severity of symptoms during the test were evaluated every 5 min using a Likert scale. We found that patients with dyspeptic symptoms after fundoplication had a significantly lower drinking capacity and higher symptoms scores than controls (P<0.05) but values similar to those of functional dyspepsia patients (P>0.05). In contrast, drinking capacity in patients without symptoms after surgery was similar to that in controls (P>0.05). These findings suggest that, as in functional dyspepsia, severe dyspeptic symptoms after Nissen fundoplication are associated with an impaired drinking capacity, reflecting visceral hypersensitivity or impaired gastric accommodation or both. PMID- 17406837 TI - Lack of effect of acute enteral arginine infusion on whole-body and intestinal protein metabolism in humans. AB - Arginine is a conditionally essential amino acid and exerts anabolic effects. We studied the effects of enteral arginine on whole-body and duodenal protein metabolism. Eight healthy fasted volunteers received randomly a 5-hr enteral infusion of either arginine (Arg; 20 g) or an isonitrogenous amino acid mixture (AA) and an IV infusion of [13C]leucine. Duodenal biopsies were performed. Whole body protein turnover and duodenal protein synthesis (FSR) were calculated from GC/MS-assessed enrichment. The mRNA levels for major components of proteolytic pathways, ubiquitin, cathepsin D, and m-calpain, were evaluated by RT-PCR. Results were compared using paired Wilcoxon test. Endogenous, oxidative, and nonoxidative leucine fluxes were not different after Arg and AA infusions, respectively. Duodenal mucosal protein FSR (71% +/- 26% vs 81% +/- 30%/day) and mRNA levels of ubiquitin, cathepsin D, and m-calpain were also similar after Arg and AA infusions. We conclude that in healthy subjects, arginine infusion exerts no effect on whole-body and duodenal protein metabolism. Whether arginine might specifically affect these parameters in catabolic or inflammatory situations remains to be determined. PMID- 17406838 TI - Morphometric analysis of small-bowel mucosa in Turkish children with celiac disease and relationship with the clinical presentation and laboratory findings. AB - We aimed to analyze morphometric features of the small-bowel mucosa in children with celiac disease, to assess the diagnostic limit values of morphometric findings, and to examine the association of morphometric findings with the clinical presentation and laboratory findings. The study comprised 33 patients with celiac disease and 35 pediatric patients undergoing endoscopy for other causes. Biopsy specimens were reanalyzed for (1) intraepithelial lymphocytes, (2) goblet cells, (3) villous height, and (4) villous/crypt ratio. The morphometric parameters of the patients were compared with controls. Then celiac patients were divided into two groups according to the presence of total villous atrophy and clinical and laboratory findings were compared. Histologic examination revealed that goblet cells, villus height, and villous/crypt ratio were significantly lower and intraepithelial lymphocytes were significantly higher in celiac patients. Cutoff values for intraepithelial lymphocytes and goblet cells in celiac patients were 31/100 and 7.8/100 epithelial cells, respectively. Moreover, for villus height and villous/crypt ratio, cutoff values were 633 microm and 0.72, respectively. Serum folic acid and vitamin B(12) levels were significantly lower in patients with total villous atrophy and were positively correlated with the severity of villous atrophy. We suggest that morphologic examination and laboratory data are important for definitive diagnosis. Villous/crypt ratio is the most sensitive and specific parameter, and intraepithelial lymphocytes may be used along with villous/crypt ratio, especially in the early phase. Folic acid and vitamin B(12) levels are good indicators of villous atrophy. PMID- 17406839 TI - Pyogenic liver abscess presenting after malignant polypectomy. AB - Streptococcus bovis bacteremia has been linked to the presence of occult colon cancer since 1977. We present a case of pyogenic liver abscess and bacteremia with a different Streptococcus viridans 1 week after colonic adenocarcinoma was removed via polypectomy, discuss the likely etiology and review whether there is evidence to support looking for colon cancer in patients who present similarly but have not already undergone screening. PMID- 17406840 TI - Pericentral liver cell necrosis associated with the use of high-dose intravenous methylprednisolone. PMID- 17406841 TI - Mucormycosis presenting as recurrent gastric perforation in a patient with Crohn's disease on glucocorticoid, 6-mercaptopurine, and infliximab therapy. PMID- 17406842 TI - Polymeric formula has direct anti-inflammatory effects on enterocytes in an in vitro model of intestinal inflammation. AB - Exclusive enteral nutrition using polymeric formula (PF) is a well-established therapeutic option for active Crohn's disease; however, its mechanisms of action are unknown. We investigated the anti-inflammatory effects of PF in an in vitro model of epithelial cell inflammation. PF did not affect cell viability over a range of dilutions, but when PF was added to the culture medium the interleukin (IL)-8 response to proinflammatory stimuli was significantly reduced. This effect was due to PF acting directly on the cells as the IL-8 response was still reduced when PF was separated from the proinflammatory stimuli in a 2-compartment system. In the presence of PF, nuclear factor (NF)-kappaB nuclear migration was not inhibited; however, IkappaBalpha degradation was delayed. PF has direct anti inflammatory effects upon immortalized colonic enterocytes. Therefore PF may, in part, modulate gut inflammation by directly reducing the inflammatory response of the intestinal epithelium. PMID- 17406843 TI - Expression of angiotensin II receptor type 1 is reduced in advanced rat liver fibrosis. AB - In this study, we assessed the hypothesis that the expression of angiotensin II receptor type 1 (AGTR1) in liver tissue changes with increasing fibrosis, which would influence the antifibrotic efficacy of AGTR1 blockers. Rats were treated with candesartancilexetil (CAN) initiated 8 or 15 days after bile duct occlusion (BDO). Four weeks after BDO, AGTR1 mRNA and protein were decreased compared to those in sham-operated animals depending on the amount of fibrosis. Starting CAN early, but not late, reduced mRNA of profibrotic TGF-beta, MMP2, and Smad2. However, CAN had no significant effect on collagen I, fibrosis, or intrahepatic resistance. In conclusion, progression of liver fibrosis reduces AGTR1 expression. Therefore, in our model, antifibrotic effects of CAN are insufficient to improve fibrosis or intrahepatic resistance. However, if AGTR1 blockade is started early, a decrease in essential profibrotic molecules is achieved. Hence, early initiation of therapy with AGTR1 blockers may be crucial for the prevention of cirrhosis. PMID- 17406844 TI - [Coblations-tonsillectomy]. PMID- 17406845 TI - [Treatment of arterial hypertension]. AB - Arterial hypertension is the most common internal disease. Treatment is highly effective in lowering cardiovascular morbidity and mortality and is indicated based on total cardiovascular risk as assessed by all relevant risk factors. Target blood pressure is <140/90 mmHg, or with concomitant diabetes mellitus or renal insufficiency <130/80 mmHg. Lifestyle modifications are helpful, either alone or as an adjuvant to drug treatment, depending on the severity of the disease. First-line drugs are diuretics, calcium antagonists, ACE inhibitors, AT1 blockers and beta blockers. In most cases, combination therapy is appropriate. Possible treatment strategies include stepped care, initial low-dose combination therapy and sequential monotherapy. PMID- 17406846 TI - The attractiveness fragment--AFLP analysis of local adaptation and sexual selection in a caeliferan grasshopper, Chorthippus biguttulus. AB - Genetic variability among males is a necessary precondition for the evolution of female choice based on indirect genetic benefits. In addition to mutations and host-parasite cycles, migration of locally adapted individuals offers an explanation for the maintenance of genetic variability. In a previous study, conducting a reciprocal transplant experiment on a grasshopper, Chorthippus biguttulus, we found that environmental conditions significantly influenced not only body condition but also an important trait of male calling song, the amplitude of song. Although not significant, all other analysed physical and courtship song traits and attractiveness were superior in native than in transferred males. Thus, we concluded that local adaptation has a slight but consistent influence on a range of traits in our study populations, including male acoustic attractiveness. In our present study, we scanned male grasshoppers from the same two populations for amplification fragment length polymorphism (AFLP) loci connected with acoustic attractiveness to conspecific females. We found greater differences in allele frequencies between the two populations, for some loci, than are expected from a balance between drift and gene flow. These loci are potentially connected with locally adapted traits. We examined whether these alleles show the proposed genotype environment interaction by having different associations with attractiveness in the two populations. One locus was significantly related to sexual attractiveness; however, this was independent of the males' population affiliation. Future research on the evolution of female choice will benefit from knowledge of the underlying genetic architecture of male traits under intraspecific sexual selection, and the 'population genomics' approach can be a powerful tool for revealing this structure. PMID- 17406847 TI - Daytime light intensity affects seasonal timing via changes in the nocturnal melatonin levels. AB - Daytime light intensity can affect the photoperiodic regulation of the reproductive cycle in birds. The actual way by which light intensity information is transduced is, however, unknown. We postulate that transduction of the light intensity information is mediated by changes in the pattern of melatonin secretion. This study, therefore, investigated the effects of high and low daytime light intensities on the daily melatonin rhythm of Afro-tropical stonechats (Saxicola torquata axillaris) in which seasonal changes in daytime light intensity act as a zeitgeber of the circannual rhythms controlling annual reproduction and molt. Stonechats were subjected to light conditions simulated as closely as possible to native conditions near the equator. Photoperiod was held constant at 12.25 h of light and 11.75 h of darkness per day. At intervals of 2.5 to 3.5 weeks, daytime light intensity was changed from bright (12,000 lux at one and 2,000 lux at the other perch) to dim (1,600 lux at one and 250 lux at the other perch) and back to the original bright light. Daily plasma melatonin profiles showed that they were linked with changes in daytime light intensity: Nighttime peak and total nocturnal levels were altered when transitions between light conditions were made, and these changes were significant when light intensity was changed from dim to bright. We suggest that daytime light intensity could affect seasonal timing via changes in melatonin profiles. PMID- 17406848 TI - Cluster formation in liverwort-associated methylobacteria and its implications. AB - Pink-pigmented methylotropic bacteria of the genus Methylobacterium inhabit the surfaces of plant organs. In bryophytes, these methylobacteria enhance cell growth, but the nature of this plant-microbe interaction is largely unknown. In this study, methylobacteria were isolated from the upper surface of the free living thalli of the liverwort Marchantia polymorpha L. Identification of one strain by 16S ribosomal RNA (rRNA) gene-targeted polymerase chain reaction (PCR) and other data show that these microbes represent an undescribed species of the genus Methylobacterium (Methylobacterium sp.). The growth-promoting activity of these wild-type methylobacteria was tested and compared with that of the type strain Methylobacterium mesophilicum. Both types of methylobacteria stimulated surface expansion of isolated gemmae from Marchantia polymorpha by about 350%. When suspended in water, the liverwort-associated bacteria (Methylobacterium sp.) formed dense clusters of up to 600 cells. In liquid cultures of Methylobacterium mesophilicum, single cells were observed, but no clustering occurred. We suggest that the liverwort-associated methylobacteria are co-evolved symbionts of the plants: Cluster formation may be a behavior that enhances the survival of the epiphytic microbes during periods of drought of these desiccation-tolerant lower plants. PMID- 17406849 TI - Bizarre tubercles on the vertebrae of Eocene fossil birds indicate an avian disease without modern counterpart. AB - Remains of fossil birds with numerous bony tubercles on the cervical vertebrae are reported from the Middle Eocene of Messel in Germany and the Late Eocene of the Quercy fissure fillings in France. These structures, which are unknown from extant birds and other vertebrates, were previously described for an avian skeleton from Messel but considered a singular feature of this specimen. The new fossils are from a different species of uncertain phylogenetic affinities and show that tuberculated vertebrae have a wider taxonomic, temporal, and geographic distribution. In contrast to previous assumptions, they are no ontogenetic feature and arise from the vertebral surface. It is concluded that they are most likely of pathologic origin and the first record of a Paleogene avian disease. Their regular and symmetrical arrangement over most of the external vertebral surface indicates a systemic disorder caused by factors that do not affect extant birds, such as especially high-dosed phytohormones or extinct pathogens. PMID- 17406850 TI - Territorial meadow pipit males (Anthus pratensis; Passeriformes) become more aggressive in female presence. AB - Although mate guarding as prevention of extra-pair copulation is common among birds, evidence for aggressive behaviour involving physical contact related to mate guarding in passerines is scarce and cases of the presence of one partner directly influencing the aggressiveness of the other are lacking. We investigated the intra-specific territorial behaviour of male meadow pipits (Anthus pratensis; Passeriformes: Motacillidae) at the beginning of the breeding season by placing a pipit model accompanied by an intra-specific song playback in the territory of socially paired males and compared the responses of males whose mates were physically present during trials with those whose females were out of sight. The level of aggression of males was significantly higher in the presence of the female; half of the males in this group physically attacked the model (the most intense and risky aggressive behaviour). Physical attacks did not occur among males whose female was absent during the trial; response to the playback by most of these males was only weak. This pattern may be related to the prevention of extra-pair copulation; if the risks involved in the conflict are outweighed by potential loss of paternity, such aggressive mate guarding may pay off. The apparently overlooked effect on the territorial behaviour of a partner's passive physical presence during conflict should be further evaluated because it may be important for the design and interpretation of results of behavioural experiments. PMID- 17406851 TI - Accumulation of additive effects generates a strong photoperiod sensitivity in the extremely late-heading rice cultivar 'Nona Bokra'. AB - Many rice cultivars that originated from lower-latitude regions exhibit a strong photoperiod sensitivity (PS) and show extremely late heading under long-day conditions. Under natural day-length conditions during the cropping season in Japan, the indica rice cultivar 'Nona Bokra' from India showed extremely late heading (202 days to heading) compared to the japonica cultivar 'Koshihikari' (105 days), from Japan. To elucidate the genetic factors associated with such extremely late heading, we performed quantitative trait locus (QTL) analyses of heading date using an F(2) population and seven advanced backcross progeny (one BC(1)F(2) and six BC(2)F(2)) derived from a cross between 'Nona Bokra' and 'Koshihikari'. The analyses revealed 12 QTLs on seven chromosomes. The 'Nona Bokra' alleles of all QTLs contributed to an increase in heading date. Digenic interactions were rarely observed between QTLs. Based on the genetic parameters of the QTLs, such as additive effects and percentage of phenotypic variance explained, these 12 QTLs are likely generate a large proportion of the phenotypic variation observed in the heading dates between 'Nona Bokra' and 'Koshihikari'. Comparison of chromosomal locations between heading date QTLs detected in this study and QTLs previously identified in 'Nipponbare' x 'Kasalath' populations revealed that eight of the heading date QTLs were recognized nearby the Hd1, Hd2, Hd3a, Hd4, Hd5, Hd6, Hd9, and Hd13. These results suggest that the strong PS in 'Nona Bokra' was generated mainly by the accumulation of additive effects of particular alleles at previously identified QTLs. PMID- 17406852 TI - The BioBreeding rat diabetes model is infected with Ljungan virus. PMID- 17406853 TI - Improved vascular engraftment and function of autotransplanted pancreatic islets as a result of partial pancreatectomy in the mouse and rat. AB - AIMS/HYPOTHESIS: The few patients subjected to autotransplantation of pancreatic islets after pancreatectomy usually become normoglycaemic after using islets from the resected organ only, whereas allogeneic recipients usually require at least two grafts to retain normoglycaemia. Previous experimental studies have demonstrated that islets transplanted to non-pancreatectomised recipients acquire a markedly decreased blood vessel density, which leads to a hypoxic microenvironment. The aim of the present study was to test the hypothesis that autotransplanted islets have better vascular engraftment and function as a result of the pancreatic surgery involved. MATERIALS AND METHODS: In the present study, athymic mice and inbred rats were subjected to a 60% pancreatectomy and transplanted with human or rat islets, respectively, 4 days later. Control animals underwent sham surgery. Blood flow, oxygen tension, vascular density and endocrine volume in the islet grafts were measured 1 month after transplantation. Separate grafts were used for perfusion experiments and for assessment of beta cell proliferation and endocrine cellular apoptosis at different time periods after transplantation. RESULTS: Islet grafts in partially pancreatectomised recipients had an increased blood flow, oxygen tension, blood vessel density and endocrine mass 1 month post-transplantation compared with control animals. They also exhibited increased insulin release in perfusion experiments performed 1 month post-transplantation, and decreased cellular apoptosis early after transplantation. CONCLUSIONS/INTERPRETATION: The present study shows that the pancreatectomy procedure itself has beneficial effects on the engraftment of transplanted human and rat islets. Our results provide an additional explanation, besides diminished immunological responses, of the much better outcome of islet autotransplantations compared with allogeneic transplantations in the clinic. PMID- 17406854 TI - Relationship between increased relative birthweight and infections during pregnancy in children with a high-risk diabetes HLA genotype. AB - AIMS/HYPOTHESIS: Children with high-risk type 1 diabetes HLA genotype have increased risk of high relative birthweight (HrBW), while cord blood islet autoantibodies decrease the risk. As gestational infections may affect offspring type 1 diabetes risk, the aims were to test whether: (1) children of mothers reporting gestational infections have increased HrBW; (2) gestational infections explain islet autoantibody reduction of HrBW; and (3) gestational infections affect the association between HLA and HrBW. SUBJECTS AND METHODS: HLA genotypes and autoantibodies to glutamic acid decarboxylase, insulinoma-associated protein 2 and insulin were determined in cord blood of children born to non-diabetic mothers in the Diabetes Prediction in Skane (DiPiS) study. Mothers reported gestational infections when the child was 2 months old. RESULTS: Fever or gastroenteritis during pregnancy was reported by 2,848/19,756 mothers (14%); 339 in more than one trimester. Children whose mothers reported infections had increased risk of HrBW (p = 0.0003), particularly in the absence of cord blood islet autoantibodies (interaction between HrBW, islet autoantibodies and infections, p = 0.0005). The effect on HrBW by high-risk HLA-DQ2/8 was aggravated by infections in more than one trimester (odds ratio [OR] = 5.24; p = 0.003) (interaction; p = 0.022). When infections were reported, cord blood islet autoantibodies decreased HrBW (OR = 0.34; p = 0.0002). CONCLUSIONS/INTERPRETATION: This study revealed that: (1) gestational fever, gastroenteritis, or both, increased the risk of HrBW; (2) cord blood islet autoantibodies decreased the risk of HrBW only in combination with infections; and (3) infections aggravated the association between HLA-DQ2/8 and HrBW. These data suggest an interaction between HLA, gestational infections, islet autoantibodies and fetal growth. PMID- 17406855 TI - [Indications and alternatives for arthroplasty in young patients]. AB - Due to the significant risk of aseptic implant loosening, the indications for endoprosthetic treatment of young adults must be assessed critically. All conservative and surgical therapy options must always be considered cautiously. Even advanced osteoarthritis of the upper extremities (shoulder, elbow and wrist joints) can often be treated with joint sparing, non-endoprosthetic therapy that leads to good clinical results and sufficient joint function. Total joint replacement is often inevitable for regaining an acceptable gait and adequate mobility in cases of advanced joint destruction of the lower extremities (hip and knee joints). Arthrodesis of the upper ankle joint in cases of isolated osteoarthritis remains a valid therapeutic option. Replacement of the upper ankle joint should be considered in cases of bilateral affliction as well as in the event of additional osteoarthritis of the adjacent joints of the lower ankle and the tarsus. PMID- 17406856 TI - Dual effect of organic acids as a function of external pH in Oenococcus oeni. AB - In this study we analyzed under various pH conditions including low pH, the effects of L-malic acid and citric acid, combined or not, on the growth, the proton motive force components and the transcription level of selected genes of the heterolactic bacterium Oenococcus oeni. It is shown here that L-malate enhanced the growth yield at pH equal or below 4.5 while the presence of citrate in media led to a complete and unexpected inhibition of the growth at pH 3.2. Nevertheless, whatever the growth conditions, both L-malate and citrate participated in the enhancement of the transmembrane pH gradient, whereas the membrane potential decreased with the pH. These results suggested that it was not citrate that was directly responsible for the inhibition observed in cultures done at low pH, but probably its end products. This was confirmed since, in media containing L-malate, the addition of acetate substantially impaired the growth rate of the bacterium and slightly the membrane potential and pH gradient. Finally, study of the expression of genes involved in the metabolism of organic acids showed that at pH 4.5 and 3.2 the presence of L-malate led to an increased amount of mRNA of mleP encoding a malate transporter. PMID- 17406857 TI - Methylphenidate reduces impulsive behaviour in juvenile Wistar rats, but not in adult Wistar, SHR and WKY rats. AB - RATIONALE: Impulsivity is a core symptom of attention deficit/hyperactivity disorder (ADHD). The spontaneously hypertensive rats (SHR) is a strain commonly used as an animal model of ADHD. However, there is no clear evidence that psychostimulants, which are used for treatment of ADHD, reduce impulsivity in SHR. Because ADHD mainly affects children, it may be relevant to study psychostimulants on juvenile animals. OBJECTIVES: Using tolerance to delay of reward as index of impulsivity, the effects of methylphenidate were assessed in adult SHR, Wistar Kyoto (WKY) and Wistar rats and in juvenile Wistar rats. MATERIALS AND METHODS: Animals were trained in a T-maze to choose between a small but-immediate and a large-but-delayed reward. Adult SHR, WKY and Wistar rats were compared for their ability to tolerate a 15-s delay. The effect of methylphenidate on the tolerance to a 30-s delay was studied in adult rats of the three strains and in juvenile (4.5 to 6.5-week-old) Wistar rats. RESULTS: In adult rats, the waiting ability was lower in SHR than in control strains. Waiting ability was improved by methylphenidate (3 and 5 mg/kg) in juveniles, but not by methylphenidate (3 mg/kg) in adults. CONCLUSIONS: These data support the idea that SHR are more impulsive than control strains. However, at the dose studied, methylphenidate fails to improve tolerance to delay in adult rats whatever the strain used. The reduction of impulsivity induced by methylphenidate in juvenile Wistar rats indicates that juvenile animals may be suitable for testing the therapeutic potential of drugs intended to the treatment of ADHD in children. PMID- 17406858 TI - Memantine produces modest reductions in heroin-induced subjective responses in human research volunteers. AB - RATIONALE: Previous studies have demonstrated an interaction between opioids and noncompetitive antagonists at N-methyl-D: -aspartate (NMDA) receptors, but few studies have examined the utility of these medications for treating opioid dependence. OBJECTIVE: In this 8-week inpatient study, participants were maintained on the low-affinity, noncompetitive NMDA receptor antagonist memantine (0, 30, and 60 mg per day, PO) and under each maintenance dose condition, the effects of intranasal heroin (0, 12.5, and 50 mg, IN) were examined. METHODS: During the first week after admission to the hospital, participants were detoxified from heroin. All of the volunteers received all of the memantine and heroin dose combinations. Participants (N = 8) first sampled a dose of heroin and $20. During a subsequent choice session, participants could self-administer heroin and/or money. Responses, which consisted of finger presses on a computer mouse, were made under a modified progressive ratio schedule (PR 50, 100, 200, 400, 800, 1,200, 1,600, 2,000, 2,400, and 2,800) during a ten-trial self administration task. Subjective, performance, and physiological effects were measured repeatedly during laboratory sessions. RESULTS: Memantine produced modest reductions in subjective ratings of drug quality, liking, willingness to pay for the drug, and craving for heroin. However, memantine produced few changes in the reinforcing effects of heroin. CONCLUSIONS: These data demonstrate that memantine was well tolerated and modestly effective in reducing the subjective but not the reinforcing effects of heroin. Although it is unlikely that memantine will be useful as a stand-alone maintenance medication for opioid dependence, it may have some utility as an adjunct treatment medication. PMID- 17406859 TI - Reversal of PCP-induced learning and memory deficits in the Morris' water maze by sertindole and other antipsychotics. AB - RATIONALE: In humans, the N-methyl-D-aspartate antagonist phencyclidine (PCP) induces behavioral changes that mimic schizophrenia symptoms, including positive and negative symptoms as well as cognitive deficits. In clinic, the cognitive deficits are closely associated with functional outcome. Thus, improvement of cognition may have high impact on patients' daily life. OBJECTIVE: In the present study, three second-generation antipsychotics (sertindole, risperidone, and clozapine) as well as the classical antipsychotic haloperidol were tested for the ability to reverse PCP-induced cognitive deficits in the Morris' water maze. RESULTS: The second-generation antipsychotics reversed the PCP-induced cognitive impairment: sertindole (0.63-2.5 mg/kg, s.c.), risperidone (0.04 mg/kg, s.c.; whereas 0.08 and 0.16 mg/kg were without significant effect), and clozapine (0.63 mg/kg, s.c.; while 1.3 mg/kg was without significant effect). The significant effect of sertindole was observed from day 2 onwards, while clozapine and risperidone only had significant effect at day 3. The classical antipsychotic haloperidol (0.010-0.020 mg/kg, s.c.) was ineffective. No compounds influenced swimming speed at the doses used, indicating that motor function was preserved. CONCLUSION: These results confirm that repeated PCP administration induces marked cognitive deficits. Further, second-generation antipsychotics like sertindole, clozapine, and risperidone within a certain, often narrow, dose range are able to reverse the impairment and thus might improve cognitive deficits in schizophrenic patients, whereas classical compounds like haloperidol lack this effect. The receptor mechanisms involved in the reversal of PCP's disruptive effect are discussed and likely include a delicate balance between effects on dopamine D(2), 5-HT(2A/6), alpha-adrenergic, muscarinic, and histaminergic H(1) receptors. PMID- 17406860 TI - The effects of self-reinnervation of cat medial and lateral gastrocnemius muscles on hindlimb kinematics in slope walking. AB - The aim of this study was to investigate the effects of self-reinnervation of the medial (MG) and lateral gastrocnemius (LG) muscles on joint kinematics of the whole hindlimb during overground walking on surfaces of varying slope in the cat. Hindlimb kinematics were assessed (1) with little or no activity in MG and LG (short-term effects of self-reinnervation), and (2) after motor function of these muscles was presumably recovered but their proprioceptive feedback permanently disrupted (long-term effects of self-reinnervation). The stance phase was examined in three walking conditions: downslope (-50%, i.e. -26.6 degrees ), level (0%) and upslope (+50%, +26.6 degrees ). Measurements were performed prior to and at consecutive time points (between 1 and 57 weeks) after transecting and immediately suturing MG and LG nerves. It was found that MG-LG self-reinnervation did not significantly change hip height and hindlimb orientation in any of the three walking conditions. Substantial short-term effects were observed in the ankle joint (e.g., increased flexion in early stance) as well as in metatarsophalangeal and knee joints, leading to altered interjoint coordination. Hindlimb kinematics in level and upslope walking progressed back towards baseline within 14-19 weeks. Thus in these two conditions the cats were walking without any detectable kinematic deficits, despite the absence of length feedback from two major ankle extensors. This was verified in a decerebrate preparation for four of the five cats. In contrast, ankle joint kinematics as well as interjoint coordination in downslope walking gradually progressed towards, but never reached their baseline patterns. The short-term effects can be explained by both mechanical and neural factors that are affected by the functional elimination of MG and LG. Permanent changes in kinematics during downslope walking indicate the importance of proprioceptive feedback from the MG and LG muscles in regulating locomotor activity of ankle extensors. Full recovery of hindlimb kinematics during level and upslope walking suggests that the proprioceptive loss is compensated by other sensory sources (e.g. cutaneous receptors) or altered central drive. PMID- 17406861 TI - Wild type and H43Y variant of human TRIM5alpha show similar anti-human immunodeficiency virus type 1 activity both in vivo and in vitro. AB - Polymorphisms in human genes have been shown to affect the rate of disease progression to acquired immune deficiency syndrome in human immunodeficiency virus type 1 (HIV-1)-infected individuals. Recently, tripartite motif 5alpha (TRIM5alpha) was identified as a factor that confers resistance to HIV-1 infection in Old World monkey cells. Subsequently, Sawyer et al. (Curr Biol 16:95 100, 2006) reported a single nucleotide polymorphism (H43Y) in the human TRIM5alpha gene and TRIM5alpha protein with 43Y was found to lose its ability to restrict HIV-1. In the present study, we reevaluated effects of this allele on in vitro anti-HIV-1 activity as well as on HIV-1 disease progression in European and Asian cohorts of HIV-1-infected individuals. Our epidemiological and molecular biological findings clearly indicate H43Y has a very minor effect on anti-HIV-1 activity of TRIM5alpha, suggesting that this allele is immaterial, at least in HIV-1-infected Europeans and Asians. PMID- 17406862 TI - Molecular characterization of MHC class II in a nonmodel anuran species, the fire bellied toad Bombina bombina. AB - While the anuran Xenopus comprises one of the best characterized nonmammalian taxa regarding the major histocompatibility complex (MHC), the organization of this gene complex has never been studied in other anurans, and information on amphibian MHC (other than Xenopus) is generally very scarce. Here, we describe the characterization of the first MHC class II B cDNA sequences from a nonmodel anuran species, the European fire-bellied toad (Bombina bombina). We isolated two transcript sequences differing substantially in amino acid composition and length within the beta2 domain. To investigate the variability of the peptide binding region in this species, we sequenced a 158-bp large fragment from wild B. bombina (n = 20) and identified eight distinct alleles. All substitutions but one were nonsynonymous, and many of the highly polymorphic sites corresponded with amino acid positions known to be involved in antigen binding. The level of variation we found in B. bombina was similar compared to that previously found in a comparable sample of a wild urodelan species, Ambystoma tigrinum, and to that found in Xenopus laevis. Based on the cDNA data and the individual's allelic diversity, we conclude that Bombina possesses at least two class II B loci. With our new beta1 primers, we were able to generate sequences in other species of anurans. We provide here a first phylogenetic analysis of this gene in amphibians. PMID- 17406863 TI - TM1 and TM2: two mutant alleles that constitute a genetic trait controlling thymocyte development. AB - Transgene insertion is instrumental to identifying genes with defined physiological functions. In this paper, we show that mice homozygous for either TM1 or TM2, two mutant alleles with distinct transgene insertions, exhibited embryo lethality, suggesting that these two alleles play essential roles in embryogenesis. Interestingly, although hemizygous TM1 or TM2 alone did not have obvious alteration in thymocyte development, together, they exhibited a compound effect on thymocyte development, blocking the development from CD4 and CD8 double negative to double-positive stage of T cells. TM1 and TM2 mutations were mapped to chromosome regions 7E-F1 and 11B5-C, respectively, where we could not identify any known gene that was implicated in a similar function. Thus, TM1 and TM2 represent two novel alleles that define a genetic trait controlling thymocyte development. PMID- 17406864 TI - Development of yeast cells displaying Candida antarctica lipase B and their application to ester synthesis reaction. AB - We isolated the lipase B from Candida antarctica CBS 6678 (CALB CBS6678) and successfully constructed CALB-displaying yeast whole-cell biocatalysts using the Flo1p short (FS) anchor system. For the display of CALB on a yeast cell surface, the newly isolated CALB CBS6678 exhibited higher hydrolytic and ester synthesis activities than the well-known CALB, which is registered in GenBank (Z30645). A protease accessibility assay using papain as a protease showed that a large part of CALB, approximately 75%, was localized on an easily accessible part of the yeast cell surface. A comparison of the lipase hydrolytic activities of yeast whole cells displaying only mature CALB (CALB) and those displaying mature CALB with a Pro region (ProCALB) revealed that mature CALB is preferable for yeast cell surface display using the Flo1p anchor system. Lyophilized yeast whole cells displaying CALB were applied to an ester synthesis reaction at 60 degrees C using adipic acid and n-butanol as substrates. The amount of dibutyl adipate (DBA) produced increased with the reaction time until 144 h. This indicated that CALB displayed on the yeast cell surface retained activity under the reaction conditions. PMID- 17406865 TI - Quantitation, regional vulnerability, and kinetic modeling of brain glucose metabolism in mild Alzheimer's disease. AB - PURPOSE: To examine CMRglc measures and corresponding glucose transport (K1 and k2) and phosphorylation (k3) rates in the medial temporal lobe (MTL, comprising the hippocampus and amygdala) and posterior cingulate cortex (PCC) in mild Alzheimer's disease (AD). METHODS: Dynamic FDG PET with arterial blood sampling was performed in seven mild AD patients (age 68+/-8 years, four females, median MMSE 23) and six normal (NL) elderly (age 69+/-9 years, three females, median MMSE 30). Absolute CMRglc (micromol/100 g/min) was calculated from MRI-defined regions of interest using multiparametric analysis with individually fitted kinetic rate constants, Gjedde-Patlak plot, and Sokoloff's autoradiographic method with population-based rate constants. Relative ROI/pons CMRglc (unitless) was also examined. RESULTS: With all methods, AD patients showed significant CMRglc reductions in the hippocampus and PCC, and a trend towards reduced parietotemporal CMRglc, as compared with NL. Significant k3 reductions were found in the hippocampus, PCC and amygdala. K1 reductions were restricted to the hippocampus. Relative CMRglc had the largest effect sizes in separating AD from NL. However, the magnitude of CMRglc reductions was 1.2- to 1.9-fold greater with absolute than with relative measures. CONCLUSION: CMRglc reductions are most prominent in the MTL and PCC in mild AD, as detected with both absolute and relative CMRglc measures. Results are discussed in terms of clinical and pharmaceutical applicability. PMID- 17406866 TI - The new EANM paediatric dosage card. AB - INTRODUCTION: In a recent publication, Jacobs et al. proposed the use of three tracer-dependent dosage cards for paediatric nuclear medicine. MATERIALS AND METHODS: Based upon this work, the EANM dosimetry and paediatrics committees introduce a condensed and revised version of this dosage card for major nuclear medicine paediatric diagnostic procedures, replacing the previous card by Piepsz et al. and including a set of minimum activities. RESULTS: The activities to be administered result in weight-independent effective doses to the children. In addition, the introduction of minimum activities guarantees a minimum standard of image quality throughout Europe and avoids a variety of administered activities in children of the same weight in different countries, which was the case when using the previous EANM dosage card. PMID- 17406867 TI - Alemtuzumab plus CHOP as front-line chemotherapy for patients with peripheral T cell lymphomas: a phase II study. AB - PURPOSE: The present study was conducted to evaluate the safety and efficacy of alemtuzumab plus CHOP chemotherapy for patients with peripheral T-cell lymphomas (PTCLs). PATIENTS AND METHODS: Twenty patients with newly diagnosed PTCLs were enrolled. The treatment consisted of classical CHOP plus alemtuzumab (10 mg i.v. on day 1 and 20 mg i.v. on day 2 in the first cycle, then 30 mg i.v. on day 1 in the subsequent cycles) based on 3-week intervals. RESULTS: Thirteen complete responses (65.0%) and three partial responses (15.0%) were confirmed, giving an overall response rate of 80.0%. The estimated event-free survival at 1 year was 43.3%. The most severe hematologic adverse event was neutropenia, which occurred with a grade-4 intensity in 18 patients (90.0%). Also, febrile neutropenia was observed in 11 patients (55.0%). Five patients (25%) experienced cytomegalovirus (CMV) reactivation, while three patients developed CMV diseases, such as pneumonitis or retinitis. There were two treatment-related deaths. Based on the high incidence of the adverse infectious and hematologic events, the current study was closed after 20 of the planned 43 patients had been enrolled. CONCLUSION: The alemtuzumab plus CHOP chemotherapy seemed to produce active antitumor activity in terms of the complete response rates in patients with PTCLs. However, since high infectious and hematologic toxicities were observed, careful monitoring and early treatment are needed to prevent treatment-related mortality. PMID- 17406868 TI - Genetic linkage of UGT1A7 and UGT1A9 polymorphisms to UGT1A1*6 is associated with reduced activity for SN-38 in Japanese patients with cancer. AB - PURPOSE: The phenotypic effects of UGT1A7 and UGT1A9 genetic polymorphisms on the in vivo pharmacokinetics of irinotecan were examined. METHODS: Eighty-four Japanese patients with cancer who received irinotecan-based chemotherapy were enrolled. Polymorphisms present in UGT1A7 (T to G transversion at -57 and UGT1A7*2 to *9), UGT1A9 (9 or 10 repeat of T at -118 [-118(T)9 or 10] and UGT1A9*2 to *5), and UGT1A1 (UGT1A1*6, UGT1A1*27, and UGT1A1*28) were analyzed for all patients. Pharmacokinetics of irinotecan were examined in 52 patients. RESULTS: The most frequent haplotype (haplotype I, 56.7%, 95% CI 53.1-60.4) consisted of polymorphisms related to normal catalytic or transcriptional activity [T at -57 and *1 of UGT1A7, -118(T)10 of UGT1A9, and UGT1A1*1]. The second most frequent haplotype (haplotype II, 15.0%, 95% CI 12.4-18.3) consisted of polymorphisms related to reduced catalytic or transcriptional activity [-57T > G and *3 of UGT1A7 and -118(T)9 of UGT1A9 linked to UGT1A1*6]. The AUC(SN 38)/AUC(SN-38G) ratios in three patients homozygous for haplotype II were significantly higher than those in 20 patients with I/I diplotype (P = 0.011). Neither of these patients had UGT1A1*28. CONCLUSION: Genetic linkage of UGT1A7 and UGT1A9 polymorphisms to UGT1A1*6, related to reduced catalytic and transcriptional activities of UGTs, is associated with the decreased glucuronosyltransferase activity for SN-38 in Japanese patients with cancer. PMID- 17406869 TI - The Aspergillus nidulans pkcA gene is involved in polarized growth, morphogenesis and maintenance of cell wall integrity. AB - The protein kinase C (PKC) family participates in maintaining integrity and growth of fungal cell walls. However, the precise molecular role of these proteins in the filamentous fungi remains unknown. In this work, pkcA, the gene encoding the PKC homolog in the filamentous fungus Aspergillus nidulans, was cloned and its function analyzed using a conditional alcA-PKC mutant strain. Repression of pkcA expression resulted in increased conidial swelling, decreased rates of hyphal growth, changes in the ultrastructure of the cell wall and increased sensitivity to antifungal agents. These results suggest that the protein encoded by pkcA is involved in key aspects of cell morphogenesis and cell wall integrity. PMID- 17406870 TI - Seronegative Lyme arthritis. AB - We present a 10-year-old girl who had been diagnosed with juvenile idiopathic arthritis 5 years before and who experienced a flare of arthritis affecting one knee while she was off medication for almost 3 years. Seronegative Lyme arthritis had to be diagnosed based on the detection of Borrelia burgdorferi DNA in synovial fluid. No humoral immune response to Borrelia burgdorferi was detectable before, at the time of diagnosis and up to 3 years later. PMID- 17406871 TI - A strong constitutive gene expression system derived from ibAGP1 promoter and its transit peptide. AB - To develop a strong constitutive gene expression system, the activities of ibAGP1 promoter and its transit peptide were investigated using transgenic Arabidopsis and a GUS reporter gene. The ibAGP1 promoter directed GUS expression in almost entire tissues including rosette leaf, inflorescence stem, inflorescence, cauline leaf and root, suggesting that the ibAGP1 promoter is a constitutive promoter. GUS expression mediated by ibAGP1 promoter was weaker than that by CaMV35S promoter in all tissue types, but when GUS protein was targeted to plastids with the aid of the ibAGP1 transit peptide, GUS levels increased to higher levels in lamina, petiole and cauline leaf compared to those produced by CaMV35S promoter. The enhancing effect of ibAGP1 transit peptide on the accumulation of foreign protein was tissue-specific; accumulation was high in lamina and inflorescence, but low in root and primary inflorescence stem. The transit peptide effect in the leaves was maintained highly regardless of developmental stages of plants. The ibAGP1 promoter and its transit peptide also directed strong GUS gene expression in transiently expressed tobacco leaves. These results suggest that the ibAGP1 promoter and its transit peptide are a strong constitutive foreign gene expression system for transgenesis of dicot plants. PMID- 17406872 TI - Arabinogalactan-proteins stimulate the organogenesis of guard cell protoplasts derived callus in sugar beet. AB - Arabinogalactan proteins (AGPs) represent a class of proteoglycans implicated in the development and differentiation of cells and tissues both in planta and in vitro. Here we report that AGP-rich extracts isolated from media of embryogenic and non-embryogenic suspension cultures of sugar beet (Beta vulgaris L.) are able to enhance the organogenesis of guard protoplast-derived callus and to increase the number of shoots formed, in comparison to control cultures. Immunocytochemical detection of carbohydrate antigens in the extracts revealed the presence of epitopes that typify both AGP and pectin, the latter being frequently bound to AGPs or, in some cases, even contributing to the polysaccharide structure of proteoglycan molecules. The most abundant epitopes proved to be those recognized by the JIM13, LM2, and MAC207 antibodies, whereas some others could be found only in relatively small or trace amounts--these included epitopes recognized by JIM16, JIM5, and LM6. Surprisingly, the JIM4- and JIM8-binding epitopes that are expressed in the course of in vitro morphogenetic processes of many species could not be detected at all in sugar beet AGPs. This is the first report of the improvement of sugar beet protoplast-derived callus organogenesis by exogenous AGP-rich extracts, an achievement that will have great impact on the biotechnological applications of protoplast technology in this species. PMID- 17406873 TI - Analysis of remote asymmetric somatic hybrids between common wheat and Arabidopsis thaliana. AB - Callus-derived protoplasts of common wheat (Triticum aestivum L. cv. Hesheng 3) irradiated with ultraviolet light were fused by using the PEG method with cell suspension-derived protoplasts of Arabidopsis thaliana. Regenerated calli and green plants resembling that of wheat were obtained. The hybrid nature of putative calli and plants were confirmed by isozyme, random amplified polymorphic DNA and genomic in situ hybridization (GISH) analyses. GISH results indicated that 1 approximately 3 small chromosome fragments of A. thaliana were found introgression into the terminals of wheat chromosomes, forming highly asymmetric hybrids. Cytoplasmic genome tests did not show any cytoplasmic genetic materials from A. thaliana. However, variations from the normal wheat cytoplasmic genome were found, indicating recombination or rearrangement occurred during the process of somatic hybridization. The chromosome elimination in the asymmetric somatic hybridization of remote phylogenetic relationship was discussed. A miniature inverted-repeat transposable element related sequence was found by chance in the hybrids which might accompany and impact the process of somatic hybridization. PMID- 17406874 TI - Interruption of pacemaker signals is mediated by GABAergic inhibition of the pacemaker nucleus in the African electric fish Gymnarchus niloticus. AB - The wave-type African weakly electric fish Gymnarchus niloticus produces electric organ discharges (EODs) from an electric organ in the tail that is driven by a pacemaker complex in the medulla, which consists of a pacemaker nucleus, two lateral relay nuclei and a medial relay nucleus. The prepacemaker nucleus (PPn) in the area of the dorsal posterior nucleus of the thalamus projects exclusively to the pacemaker nucleus and is responsible for EOD interruption behavior. The goal of the present study is to test the existence of inhibition of the pacemaker nucleus by the PPn. Immunohistochemical results showed clear anti-GABA immunoreactive labeling of fibers and terminals in the pacemaker nucleus, but no apparent anti-glycine immunoreactivity anywhere in the pacemaker complex. GABA injection into the pacemaker nucleus could induce EOD interruptions that are comparable to the interruptions induced by glutamate injection into the PPn. Application of the GABA(A) receptor blocker bicuculline methiodide reversibly eliminated the effects of stimulation of the PPn. Thus the EOD interruption behavior in Gymnarchus is mediated through GABAergic inhibition of the pacemaker nucleus by the PPn. PMID- 17406875 TI - Localized amyloidosis in endometrioid carcinoma of the uterus: a rare association. AB - BACKGROUND: Amyloidosis is associated with infections, chronic disorders, hematological malignancies and endocrine neoplasms. Only rarely has it been associated with other tumors. Amyloidosis of the endometrium is rare and has only been reported as part of a systemic disease. CASE REPORT: A 65-year-old postmenopausal female presented with irregular vaginal bleeding. Ultrasonography revealed a growth in the uterine cavity and enlarged internal and external iliac lymph nodes. Total hysterectomy with Bilateral salpingo-ophorectomy and dissection of pelvic fascia with external and internal iliac lymph nodes was done. The histologic diagnosis was that of a well-differentiated endometrioid adenocarcinoma: villoglandular type. Additionally, there were intratumoral nodular deposits of amyloid which were positive with Congo red showing apple green birefringence on polarization. CONCLUSION: This is an unusual case of amyloidosis associated with endometrioid carcinoma of the uterus. PMID- 17406876 TI - Uterine prolapse complicating pregnancy. AB - BACKGROUND: Uterine prolapse is extremely rare during pregnancy. However in some cases significant complications may develop. We report a case of uterine prolapse which developed during pregnancy. Our case was managed conservatively and there were no fetal or maternal complications. Postnatally the uterine prolapse recovered spontaneously. Early recognition and close follow-up during pregnancy is essential. Successful pregnancy outcome requires individualized treatment but bed rest should always be considered. PMID- 17406877 TI - Neu-Laxova syndrome, grossly appearing normal on 20 weeks ultrasonographic scan, that manifested late in pregnancy: a case report. AB - INTRODUCTION: Neu-Laxova syndrome (NLS) is a very rare and extremely lethal syndrome almost always inherited by autosomal recessive pattern. CASE PRESENTATION: A 21-year-old G2 P1 L0 Turkish woman was first seen at 18 weeks of gestation in antenatal policlinic. On ultrasound examination performed at 20 weeks of gestation for excluding gross abnormalities showed no abnormality and biometry of the fetus was concordant with date of gestation. She stopped follow up after then. The woman were presented to emergency unit with a complaint of absent fetal movements at 38 weeks of gestation. Multiple abnormalities consistent with NLS have been seen on sonography. Nonvisible gastric cavity was also seen on sonography. Postmortem examination by genetics specialists confirmed the diagnosis of NLS. CONCLUSION: Neu-Laxova syndrome must be monitored continuously from the beginning of gestation to third trimester via serial ultrasonographies that is focused on previously described features of syndrome especially for consanguineous couples with or without a previous affected fetus. PMID- 17406878 TI - Galactocele as a changing axillary lump in a pregnant woman. AB - PURPOSE: We report a galactocele as changing axillary lumps in both axillae. MATERIALS AND METHODS: 36-year-old woman visited to our hospital for her axillary lump. The diagnosis could be made under ultrasound examination and her recent pregnancy history. However, sonography-guided aspiration was done for confirmation. RESULT: We could confirm the galactocele arising in axillary accessory breast tissue. CONCLUSION: Clinical information indicating that such a palpable mass is associated with pregnancy as well as its sonography features along with adjunctive diagnostic aspiration, can help to correctly diagnose this rare entity. PMID- 17406879 TI - A late post-traumatic diaphragmatic hernia revealed during pregnancy by post partum respiratory distress. AB - INTRODUCTION: Diaphragmatic hernia during pregnancy is uncommon and is usually traumatic in origin, epigastric pain, and vomiting could be the initial symptoms of herniation of gastrointestinal contents, with a risk of strangulation and ischaemia, leading to respiratory distress due to collapse of the lung. METHODS: Case report. RESULTS: A 27-year-old woman, with undiagnosed traumatic diaphragmatic hernia who presented, at 32 weeks' gestation, epigastric pain, vomiting and tachycardia, immediate post-partum course was complicated by respiratory failure. A chest X-ray showed an air fluid level in the left lung which was wrongly diagnosed as an hydropneumothorax, in front of respiratory symptoms exacerbation, an inappropriate thoracic drainage tube was accidentally placed into the herniated stomach leading to perforation of this last. An emergency laparotomy discovered a 2/3 of the stomach, transverse colon and greater omentum herniated in the left hemithorax through a defect of the left hemidiaphragm. CONCLUSION: The diagnosis should then be considered early, and chest radiography with a nasogastric tube is the first technique to prefer and may be helpful to confirm the diagnosis. PMID- 17406880 TI - Subtraction of background damage in PFGE experiments on DNA fragment-size distributions. AB - The non-random distribution of DNA breakage in pulsed-field gel electrophoresis (PFGE) experiments poses a problem of proper subtraction of the background damage to obtain a fragment-size distribution due to radiation only. As been pointed out by various authors, a naive bin-to-bin subtraction of the background signal will not result in the right DNA mass distribution histogram, and may even result in negative values. Previous more systematic subtraction methods have been based mainly on random breakage, appropriate for low-LET radiation but problematic for high LET. Moreover, an investigation is needed whether the background breakage itself is random or non-random. Previously a new generalized formalism based on stochastic processes for the subtraction of the background damage in PFGE experiments for any LET and any background was proposed, and as now applied it to a set of PFGE data for Fe ions. We developed a Monte Carlo algorithm to compare the naive subtraction procedure in artificial data sets to the result produced by the new formalism. The simulated data corresponded to various cases, involving non-random (high-LET) or random radiation breakage and random or non-random background breakage. The formalism systematically gives better results than naive bin-by-bin subtraction in all these artificial data sets. PMID- 17406881 TI - Identification and sequence analysis of discordant phenotypes between AmpFlSTR SGM Plus and PowerPlex 16. AB - During duplicate analysis of buccal swabs from 1,377 individuals with 2 commercial short tandem repeat (STR) kits, we observed 8 discordant phenotypes with SGM Plus (SGM, second generation multiplex) for the STRs THO1 (2), vWA (4) and D18S51 (2), and 1 discrepancy with PowerPlex 16 for D18S51. One individual even showed two discrepancies (vWA and THO1) for SGM Plus. In each case, the difference observed was due to the non-amplification or allele dropout of the second allele in a heterozygous genotype. Sequence analysis revealed each time the presence of a mutation that probably coincided with the primer-binding site. Primer-binding site mutations for vWA and D18S51 have been reported previously, while the mutation for THO1 (C-to-T substitution at position 1286 of GenBank sequence D00269) is reported here for the first time. While the frequency of these silent alleles remains low (0.58% in our study), it is suggested that appropriate measures should be taken for database comparisons and that allelic dropout should be further investigated by sequence analysis and be reported to the forensic community. PMID- 17406882 TI - Subtype lesions of neovascular age-related macular degeneration in Chinese patients. AB - PURPOSE: To identify the subtype frequency and clinical features of neovascular age-related macular degeneration (AMD) in Chinese patients. METHODS: From January 2003 to August 2006, we investigated prospectively 155 newly diagnosed patients with presumed neovascular AMD. Fundus fluorescein angiography (FFA) and indocyanine green angiography (ICGA) were performed in both eyes of all patients. Subtype frequency and clinical features were recorded according to their angiograms. RESULTS: Three subtypes of lesion were noted, which were polypoidal choroidal vasculopathy (PCV), retinal angiomatous proliferation (RAP) and mixed lesions. Of the 155 patients, 105 (67.7%) had choroidal neovascularization (CNV) of the typical type seen in AMD, 38 (24.5%) had PCV and seven (4.5%) had RAP. In five (3.2%) additional cases, mixed lesions were noted. In 38 cases (47 eyes) with PCV, the rates of subfoveal, juxtafoveal and extrafoveal lesion were respectively 29.8% (14 eyes), 8.5% (four eyes), and 61.7% (29 eyes), compared with 75.6%, 14.6% and 9.8% for CNV lesion (P < 0.01). The percentage of subfoveal lesion in PCV group was significantly lower than that in the CNV group (P < 0.01). The location of the RAP lesion was subfoveal in two (28.6%) eyes, juxtafoveal in three (42.9%) eyes and extrafoveal in two (28.6%) eyes. The five eyes with mixed lesions were all PCV coexisting with CNV at the same eye, and in all of the five cases, CNV was subfoveal while PCV was extrafoveal. CONCLUSIONS: In this hospital-based study, PCV accounts for 24.5% of neovascular AMD and is the most common subtype, RAP is less frequent (4.5%), and mixed lesions are much less common in Chinese patients. PCV is least likely to involve the fovea in neovascular AMD. PMID- 17406883 TI - The economic burden of diabetic retinopathy in Germany in 2002. AB - BACKGROUND: The aims of our study were to describe the costs associated with diabetic retinopathy (DR), and to evaluate its economic impact in Germany. METHODS: Forty-one German ophthalmologists, randomly selected from a physicians' database in Germany, provided information on adult Type 1 and Type 2 diabetic patients with DR (n = 207). This information included socio-demographics, clinical characteristics and resource use during the year 2002. National-level cost estimates were calculated, based on these results and the prevalence data on DR in Germany. RESULTS: This study found that costs associated with DR tend to increase as DR progresses, being highest in patients with proliferative DR and lowest in patients with mild, non-proliferative DR. The German statutory health insurance (Gesetzliche Krankenversicherung, GKV) covered two-thirds of the total costs paid by all the payers. The total cost of DR from a societal perspective was calculated at 3.51 euros bn for the year 2002, and from the GKV perspective amounted to 2.23 euros bn. CONCLUSIONS: This study is the first comprehensive study to provide estimates of costs associated with DR in Germany. These costs were estimated to account for approximately 1.5% of the total health-care expenditure in 2002. PMID- 17406884 TI - Glycoxidized particles mimic lipofuscin accumulation in aging eyes: a new age related macular degeneration model in rabbits. AB - PURPOSE: The biogenesis of drusen, a hallmark of age-related macular degeneration (AMD), is still unclear. Lipofuscin, which extensively accumulates with age in RPE cells, is hardly soluble, derived in part from oxidation byproducts of the photoreceptor outer segments. The purpose of the current study is to develop a new AMD model in rabbits using glycoxidized particles as imitation lipofuscin, and determine whether accumulation of lipofuscin as insoluble material may play a role in drusen biogenesis and other pathogenesis of AMD. METHODS: To mimic the accumulation of insoluble lipofuscin, glycoxidized microspheres (glycox-MS) were made through a glycoxidation process with albumin and glycolaldehyde, alpha hydroxy aldehyde. As a control, microspheres made with glutaraldehyde (cMS) and soluble glycoxidized (glycox-) albumin were prepared. Each material was implanted into the subretinal space in rabbits. The implanted area was assessed by funduscopy, fluorescein angiography, histology, and transmission electron microscopy (TEM). RESULTS: Compared with control microspheres, glycox-MS stagnated for a prolonged period in the cytoplasm of RPE cells. Eyes implanted with glycox-MS produced drusen-like deposits at a significantly higher frequency, when compared with the controls. Glycox-MS were observed at the margin of or beneath the drusen-like deposits in all cases. In some eyes with glycox-MS, late onset sub-RPE choroidal neovascularization was observed, while control groups did not have these findings. CONCLUSIONS: These results suggest that the accumulation of indigestible granules such as lipofuscin in RPE or subsequent depositions toward Bruch's membrane may play a role in drusen biogenesis as a trigger of inflammation or via other mechanisms. This model of AMD may be useful to elucidate drusen biogenesis and pathogenesis of AMD. PMID- 17406885 TI - Long-term effects of radial optic neurotomy for central retinal vein occlusion consecutive interventional case series. AB - PURPOSE: To investigate the long-term (minimum 24 months follow-up) clinical results of radial optic neurotomy (RON) following a pars plana vitrectomy (PPV) with internal limiting membrane peeling as treatment for central retinal vein occlusion (CRVO). METHODS: Interventional case series of 14 consecutive patients (14 eyes) with CRVO who were treated with a PPV combined with RON within 1 year of diagnosis. RESULTS: Median baseline visual acuity (VA) was 1.05 logMAR (approximately 0.09 Snellen) in the affected eye. The follow-up period ranged from 24 to 48 months postoperatively, median 30 months. At the 24-month follow-up examination, median VA was 1.005 logMAR in the affected eye-a significant improvement (p = 0.013). Six patients (43%) gained 1 or more lines of VA (mean VA gain = 1.7 lines), while the VA of four patients (29%) improved by 3 or more lines. The eyes with nonischemic CRVO demonstrated a significantly higher improvement in VA (p = 0.0007) than the eyes with ischemic CRVO. CONCLUSION: With RON clinically relevant improvements on a long-term basis seem achievable. Patients with nonischemic CRVO may respond more favorably than patients with ischemic CRVO. PMID- 17406886 TI - Endurance training improves post-exercise cardiac autonomic modulation in obese women with and without type 2 diabetes. AB - Obesity and type 2 diabetes (T2D) are associated with abnormal cardiovascular autonomic function and increased risk for cardiac complications, especially after exercise. Since improvements at rest are not always observed after training, we investigated changes in resting and post-exercise autonomic function in obese women with and without T2D after16-week of walking training. Heart rate (HR) variability (HRV) and baroreflex sensitivity (BRS) were measured at rest and 20 min after a 20 min bout of treadmill exercise at 65% VO(2) peak in obese women with (n = 8) and without T2D (n = 12) before and after training. HRV was analyzed by frequency-domain [high- (HF) power and low-frequency (LF)] and BRS by the sequence method. Exercise training induced similar significant changes in VO(2) peak, resting systolic blood pressure (SBP) and post-exercise autonomic function in both groups. Training increased VO(2 )peak (6%; P < 0.01) and decreased resting SBP (8%; P < 0.001). Increased post-exercise HR recovery (5%; P < 0.001), HF power (14%; P < 0.05), LF power (14%; P < 0.05) and BRS (86%; P < 0.001) were also observed. Resting autonomic function and post-exercise SBP were not altered after training. In conclusion, endurance training reduced blood pressure without changes in HRV and BRS at rest, but training increased HRV and BRS during the recovery of acute endurance exercise indicating an improved post-exercise autonomic modulation of HR, which was similar in obese women with and without T2D. PMID- 17406888 TI - Effects of fasting and carbohydrate consumption on voluntary resting apnea duration. AB - Breath holding is normally terminated due to the urge to breathe, mainly caused by the increasing carbon dioxide level. It was recently shown that a combination of 18 h of carbohydrate-free diet and prolonged exercise prior to breath holding lowered the respiratory exchange ratio (RER) and end-expiratory PO(2 )at maximal breath-hold break-point (MBP). Current hypothesis: fasting will result in longer breath-hold duration than will fasting followed by carbohydrate intake. It was also hypothesized that breath-holds during carbohydrate supplementation would be ended at a higher alveolar PO(2). Ten male non-divers performed multiple breath holds either to the first diaphragmatic contraction (PBP), or to MBP. The breath holds were performed during normal diet (control; C), twice during fasting (F14 h, F18 h), followed by post fasting carbohydrate consumption (PFCC) and a repetition of the breath-hold testing 1-2 h after ingestion of carbohydrates. Duration, RER, end-tidal PO(2) and PCO(2), SaO(2) and blood glucose were determined. RER and blood glucose increased after PFCC compared with fasting and control conditions (P < 0.001). PBP breath-hold duration increased from 129 +/- 34 s at C to 148 +/- 33 s at F18 h, and was reduced during PFCC to 122 +/- 30 s (P < 0.001). End-tidal PO(2) was higher at PFCC compared to F18 h (10.4 +/- 1.2 vs. 9.5 +/- 1.2 kPa at PBP, P < 0.05). Similar trends in breath-hold duration and physiology were seen in breath-holds that were terminated at MBP. Dietary restriction can affect breath-hold duration. The lower O(2) level at breakpoint during fasting suggests that breath holding may be less safe during fasting; the increased risk may be mitigated by ingestion of carbohydrates before breath holding. PMID- 17406887 TI - Vertical jump performance after 90 days bed rest with and without flywheel resistive exercise, including a 180 days follow-up. AB - Muscle atrophy and neuromuscular de-conditioning occur in response to space flight and bed-rest. In this study, we investigated the efficacy of flywheel training to conserve jumping power and height during 90 days bed rest. Twenty four young healthy men underwent strict bed-rest (-6 degrees head down tilt) for 90 days. Eight participants were assigned to a flywheel group (FW) and 16 to a control group (Ctrl). The ground reaction force was measured during vertical jump tests twice during baseline data collection, and on day 4, 7, 14, 90 and 180 of recovery. In half of the participants, jump tests were also performed within minutes after re-ambulation and on four more occasions during the first 2 days of recovery. Jump height was reduced from 40.6 cm (SD 6.1 cm) during the first baseline measurement to 27.6 cm (SD 5.6 cm) on day 4 of recovery in Ctrl, but only from 38.6 cm (SD 3.9 cm) to 34.4 cm (SD 6.5 cm) in FW (P < 0.001). At the same time, peak power was reduced from 47.4 W/kg (SD 8.0 W/kg) to 34.5 W/kg in Ctrl, but only from 46.2 W/kg (6.0 W/kg) to 42.2 W/kg SD 4.6 W/kg) in FW (P < 0.001). Jump height and peak power were completely recovered after 163 and 140 days in Ctrl, respectively, and after 72 and 18 days in FW (regression analysis). In conclusion, flywheel exercise could effectively offset neuromuscular de conditioning during bed-rest, and led to full recovery at an earlier stage. These findings nourish the hope that adequate training paradigms can fully sustain neuromuscular function under microgravity conditions. PMID- 17406889 TI - Affine differential geometry analysis of human arm movements. AB - Humans interact with their environment through sensory information and motor actions. These interactions may be understood via the underlying geometry of both perception and action. While the motor space is typically considered by default to be Euclidean, persistent behavioral observations point to a different underlying geometric structure. These observed regularities include the "two thirds power law", which connects path curvature with velocity, and "local isochrony", which prescribes the relation between movement time and its extent. Starting with these empirical observations, we have developed a mathematical framework based on differential geometry, Lie group theory and Cartan's moving frame method for the analysis of human hand trajectories. We also use this method to identify possible motion primitives, i.e., elementary building blocks from which more complicated movements are constructed. We show that a natural geometric description of continuous repetitive hand trajectories is not Euclidean but equi-affine. Specifically, equi-affine velocity is piecewise constant along movement segments, and movement execution time for a given segment is proportional to its equi-affine arc-length. Using this mathematical framework, we then analyze experimentally recorded drawing movements. To examine movement segmentation and classification, the two fundamental equi-affine differential invariants-equi-affine arc-length and curvature are calculated for the recorded movements. We also discuss the possible role of conic sections, i.e., curves with constant equi-affine curvature, as motor primitives and focus in more detail on parabolas, the equi-affine geodesics. Finally, we explore possible schemes for the internal neural coding of motor commands by showing that the equi-affine framework is compatible with the common model of population coding of the hand velocity vector when combined with a simple assumption on its dynamics. We then discuss several alternative explanations for the role that the equi-affine metric may play in internal representations of motion perception and production. PMID- 17406890 TI - Histological analysis of direct somatic embryogenesis in Arabidopsis thaliana (L.) Heynh. AB - In Arabidopsis the in vitro culture of immature zygotic embryos (IZEs) at a late stage of development, on the solid medium containing synthetic auxin, leads to formation of somatic embryos via direct somatic embryogenesis (DSE). The presented results provide evidence that in IZE cells competent for DSE are located in the protodermis and subprotodermis of the adaxial side of cotyledons and somatic embryos displayed a single- or multicellular origin. Transgenic Arabidopsis lines expressing the GUS reporter gene, driven by the DR5 and LEC2 promoters, were used to analyse the distribution of auxin to mark embryogenic cells in cultured explants and develop somatic embryos. The analysis showed that at the start of the culture auxin was accumulated in all explant tissues, but from the fourth day onwards its location shifted to the protodermis and subprotodermis of the explant cotyledons. In globular somatic embryos auxin was detected in all cells, with a higher concentration in the protodermis, and in the heart stage its activity was mainly displayed in the shoot, root pole and cotyledon primordia. The embryogenic nature of dividing protodermal and subprotodermal cells accumulating auxin was confirmed by high expression of promoter activity of LEC2 in these cells. Analysis of symplasmic tracer (CFDA) distribution indicated symplasmic isolation between tissues engaged in DSE and other parts of an explant. Symplasmic isolation of somatic embryos from the explant was also detected. PMID- 17406891 TI - Molecular cytogenetic characterization of t(14;19)(q32;p13), a new recurrent translocation in B cell malignancies. AB - Translocations involving an immunoglobulin (IG) locus are a recurring theme in B cell neoplasia. The rearrangements lead to the joining of an IG gene with a (proto)oncogene, whereby the latter comes under the influence of transcription stimulating sequences in the constitutively active IG locus resulting in deregulation of the oncogene and neoplastic growth. We present here three cases of B cell neoplasia that showed a t(14;19)(q32;p13) by karyotypic analysis. Detailed molecular cytogenetic characterization of the breakpoints on chromosomes 14 and 19 in the two cases from which extra material was available, demonstrated the involvement of the immunoglobulin heavy-chain (IGH@)-variable region on chromosome 14 in both and, in one case, that the breakpoint was within the BRD4 gene on chromosome 19. Against the background of what one knows about IGH@ involvement in lymphatic malignancies, it is difficult to envisage a fusion gene with qualitatively altered protein product as the crucial pathogenetic outcome of the translocation. In spite of the fact that we found BRD4 split by the t(14;19)(q32;p13) in one of the two informative cases, we cannot be sure that this was the pathogenetically relevant target gene. Other pathogenetic possibilities could be deregulation of the neighboring NOTCH3 and/or ABHD9 genes, located distal to BRD4 in 19p13. PMID- 17406892 TI - The spectrum of persistence of testicular blastema and ectopic testicular parenchyma: a possible result of focal delay in gonadal development. AB - Sex-cord formation and organization are important steps in testicular development and depend on adequate interactions between mesenchymal cells, pre-Sertoli cells, and germ cells. These elements form the testicular blastema, the precursor of the testicular parenchyma, morphologically characterized by poorly organized sex cords and mesenchymal components. Here, we study two uncommon testicular lesions, unrelated to other gonadal anomalies. In the first group, we describe the features of persistence of testicular blastema in three fetal autopsy cases, discussing its possible pathogenesis and clinical importance. In the second, we analyze 11 cases of ectopic testicular parenchyma in the tunica albuginea, an uncommon benign condition of uncertain clinical significance, whose main differential diagnosis is gonadal dysgenesis. Based on their similar topography within the testis, and on their possibly shared embryological origin, we propose that both lesions may represent the two extremes of a maldevelopmental spectrum resulting from a focal delay in testicular development. PMID- 17406893 TI - How often does cervical involvement upstage patients with non-myoinvasive (otherwise stage 1A) endometrioid adenocarcinoma of the endometrium? PMID- 17406894 TI - Tissue preparation for MOHS' frozen sections: a comparison of three techniques. AB - Accurate, efficient frozen section analysis is important for tumor control. A few studies address the technical issues. More are needed, especially as new technologies become available. The objective of this study is to compare the efficiency of three techniques of flattening tissue for microscopically oriented histologic surgery (MOHS): conventional frozen sectioning, Cryocup, and CryoHist. Conventional chuck/heat sink-frozen section preparation were compared with Cryocup and CryoHist to determine the most efficient technique to examine 100% of the surgical margin of 4-cm diameter, full thickness, fresh autopsy cylinders of anterior abdominal skin, which were marked on their deep and peripheral margins. The specimens were frozen sectioned at 5 microm until all the marking dye was gone from the deep surface, and 95% of the perimeter epidermis could be seen. The conventional chuck required an average of 304 micrometers to clear the deep margin and four fifths did not contain 95% of the epidermal margin. The Cryocup required an average of 284 microm to examine the deep margin and 95% of the epidermal margin. The CryoHist required an average of 104 microm to examine the deep margin and 95% of the epidermal border. The new techniques improve the efficiency and presumably the accuracy of tumor margin analysis. PMID- 17406897 TI - Catalase, Bax and p53 expression in the visual system of the crab Ucides cordatus following exposure to ultraviolet radiation. AB - In invertebrates, a few studies have suggested apoptosis as the mechanism of choice to protect the retina after exposure to ultraviolet (UV) radiation. We demonstrated previously, by electron microscopy, that the retina and lamina ganglionaris (or lamina) cells of the crab Ucides cordatus displayed subcellular signs of apoptosis after exposure to UVB and UVC. Here, we first ascertained, by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) technique, that UV irradiation indeed produced the previously reported results. We next tested, in the visual system of U. cordatus, whether the expression (as analyzed by immunohistochemistry and observed with laser scanning microscopy) and levels (as examined by Western blotting) of catalase, Bax, and p53 were affected by the same dose of UV radiation as that used previously. Our data revealed that the intensity of catalase, Bax, and p53 labeling was stronger in irradiated retina and lamina cells than in non-irradiated retina and lamina. However, no significant difference was observed in the concentrations of these proteins isolated from the whole optic lobe. The results thus suggest that UVB and UVC induce apoptosis in the crustacean retina and lamina by increasing catalase expression and activating the Bax- and p53-mediated apoptosis pathways. PMID- 17406896 TI - Leptin downregulates heat shock protein-70 (HSP-70) gene expression in chicken liver and hypothalamus. AB - Heat shock protein (HSP)-70 is expressed in normal and stressed cells but is highly stress-inducible. Although leptin has long been suggested to be involved in the regulation of stress response, its interaction with the HSP-70 gene is still unknown, under both unstressed and stressed conditions. The present study has aimed to investigate the effect of leptin on HSP-70 gene expression in normal chicken liver, hypothalamus, and muscle. Continuous infusion of recombinant chicken leptin (8 mug/kg per hour) at a constant rate of 3 ml/h for 6 h in 3-week old broiler chickens significantly (P < 0.05) decreased food intake and HSP-70 mRNA levels in liver and hypothalamus, but not in muscle. In an attempt to discriminate between the effect of leptin and of leptin-reduced food intake on HSP-70 gene expression, we also evaluated the effect of food deprivation on the same cellular responses in two broiler chicken lines genetically selected for low (LL) or high (FL) abdominal fat pad size. Food deprivation for 16 h did not affect HSP-70 gene expression in any of the studied tissues indicating that the effect of leptin was independent of the inhibition of food intake. Regardless of the nutritional status, HSP-70 mRNA levels were significantly (P < 0.05) higher in the hypothalamus of FL compared with LL chickens consistent with higher mRNA levels for hypothalamic corticotropin-releasing factor. To assess, whether the effects of leptin were direct or indirect, we carried out in vitro studies. Leptin treatments did not affect HSP-70 mRNA levels in a leghorn male hepatoma cell line or quail myoblast cell line suggesting that the effect of leptin on HSP 70 gene expression is mediated through the central nervous system. Furthermore, HSP-70 gene expression was gender-dependent with significantly (P < 0.05) higher levels in male than in female chickens. PMID- 17406895 TI - The threat of avian influenza a (H5N1): part II: Clues to pathogenicity and pathology. AB - Among emerging and re-emerging infectious diseases, influenza constitutes one of the major threats to mankind. In this review series epidemiologic, virologic and pathologic concerns raised by infections of humans with avian influenza virus A/H5N1 are discussed. The second part focuses on experimental and clinical results, which give insights in the pathogenic mechanisms of H5N1 infection in humans. H5N1 is poorly transmitted to humans. However, H5N1-induced disease is very severe. More information on the role entry barriers, H5N1 target cells and on H5N1-induced modulation of the host immune response is needed to learn more about the determinants of H5N1 pathogenicity. PMID- 17406898 TI - A proteomic approach to the investigation of early events involved in the activation of vascular smooth muscle cells. AB - Vascular smooth muscle cells (VSMC) are mature cells that maintain great plasticity. This distinctive quality is the basis of the migration and proliferation of VSMC in cardiovascular diseases. We have investigated, via a proteomic approach, the molecular changes that promote VSMC switching from a quiescent to an activated-proliferating phenotype. In particular, we focus on the modulation in tyrosine phosphorylation that occurs in cell activation by serum or by single growth factors, such as insulin-like growth factor 1 (IGF-1) or platelet-derived growth factor (PDGF-BB). A comparison of profiles from two dimensional polyacrylamide gel electrophoresis analysis of quiescent and activated-proliferating VSMC has revealed a number of differences in protein expression. Several differentially expressed proteins have been identified by mass spectrometry, and their changes during the time course of tyrosine phosphorylation have been documented from time zero up to 48 h after stimulus. The tyrosine-phosphorylation level generally decreases within a few minutes of stimulation, followed by a rapid dramatic recovery of some chaperones and redox enzymes, but no significant recovery for glucose metabolism enzymes. With respect to cytoskeleton components, no remarkable fluctuations have been detected at the earliest time points, except for those relating to alpha-actin, which displays an impressive decrease. A comparison of the early stages of cell stimulation after serum or after single growth factor administration has revealed important differences in the phosphorylation of chaperones, thereby suggesting their crucial role in VSMC activation. PMID- 17406899 TI - Expression of adhesion molecules during normal pregnancy. AB - We review the available information regarding the role of adhesive molecules as potential participants in the complex events of fertilization, embryogenesis, implantation and placentation. Studies that specifically relate to the expression and modulation of adhesive molecules in fertilization, embryogenesis, and implantation have been identified in the literature and by Medline searches. Cell cell and cell-extracellular matrix interactions play a critical role in various developmental processes and in the cascade of events that lead to implantation and to the normal development of the fetus during pregnancy. Adhesion molecules influence, directly or indirectly, numerous aspects of cell behaviour, cell migration, cell growth, cell survival, cell proliferation, angiogenesis, invasion and metastasis. PMID- 17406900 TI - Expression of PU.1 and terminal differentiation of alveolar macrophages in newborn rats. AB - PU.1, which is a transcription factor, promotes the terminal differentiation of alveolar macrophages (AMs). Its expression is regulated by granulocyte/macrophage colony-stimulating factor (GM-CSF). In this study of AMs in newborn rats, we performed immunohistochemical staining, acid phosphatase staining, reverse transcriptase polymerase chain reaction (RT-PCR), quantitative real-time PCR, cytokine assay, and electron microscopy. AMs at 3 and 7 days after birth had a large foamy appearance with an intracytoplasmic accumulation of surfactants. Weak expression of PU.1 was observed in the nuclei. AMs at 15 days after birth were smaller, and PU.1 expression had increased. Ultrastructurally, AMs at 1 day after birth had a smooth surface and abundant lamellar structures in the cytoplasm, whereas AMs at 56 days after birth were characterized by (1) abundant microvillar projections on the cell surface, and (2) well-developed lysosomes and a few lamellar structures in the cytoplasm. Acid phosphatase activity and the expression of mannose receptor, scavenger receptor, and GM-CSF receptor alpha were enhanced in AMs with time after birth. These results suggest that AMs are initially immature, and that their terminal differentiation starts after birth concomitantly with an increased expression of PU.1. PMID- 17406901 TI - Formation of giant spicules in the deep-sea hexactinellid Monorhaphis chuni (Schulze 1904): electron-microscopic and biochemical studies. AB - The siliceous sponge Monorhaphis chuni (Hexactinellida) synthesizes the largest biosilica structures on earth (3 m). Scanning electron microscopy has shown that these spicules are regularly composed of concentrically arranged lamellae (width: 3-10 mum). Between 400 and 600 lamellae have been counted in one giant basal spicule. An axial canal (diameter: ~2 mum) is located in the center of the spicules; it harbors the axial filament and is surrounded by an axial cylinder (100-150 mum) of electron-dense homogeneous silica. During dissolution of the spicules with hydrofluoric acid, the axial filament is first released followed by the release of a proteinaceous tubule. Two major proteins (150 kDa and 35 kDa) have been visualized, together with a 24-kDa protein that cross-reacts with antibodies against silicatein. The spicules are surrounded by a collagen net, and the existence of a hexactinellidan collagen gene has been demonstrated by cloning it from Aphrocallistes vastus. During the axial growth of the spicules, silicatein or the silicatein-related protein is proposed to become associated with the surface of the spicules and to be finally internalized through the apical opening to associate with the axial filament. Based on the data gathered here, we suggest that, in the Hexactinellida, the growth of the spicules is mediated by silicatein or by a silicatein-related protein, with the orientation of biosilica deposition being controlled by lectin and collagen. PMID- 17406902 TI - Study of tauopathies by comparing Drosophila and human tau in Drosophila. AB - The microtubule-binding protein tau has been investigated for its contribution to various neurodegenerative disorders. However, the findings from transgenic studies, using the same tau transgene, vary widely among different laboratories. Here, we have investigated the potential mechanisms underlying tauopathies by comparing Drosophila (d-tau) and human (h-tau) tau in a Drosophila model. Overexpression of a single copy of either tau isoform in the retina results in a similar rough eye phenotype. However, co-expression of Par-1 with d-tau leads to lethality, whereas co-expression of Par-1 with h-tau has little effect on the rough eye phenotype. We have found analogous results by comparing larval proteomes. Through genetic screening and proteomic analysis, we have identified some important potential modifiers and tau-associated proteins. These results suggest that the two tau genes differ significantly. This comparison between species-specific isoforms may help to clarify whether the homologous tau genes are conserved. PMID- 17406903 TI - Neuroimmune connections in jejunal and ileal Peyer's patches at various bovine ages: potential sites for prion neuroinvasion. AB - During preclinical stages of cattle orally infected with bovine spongiform encephalopathy (BSE), the responsible agent is confined to ileal Peyer's patches (IPP), namely in nerve fibers and in lymph follicles, before reaching the peripheral and central nervous systems. No infectivity has been reported in other bovine lymphoid organs, including jejunal Peyer's patches (JPP). To determine the potential sites for prion neuroinvasion in IPP, we analyzed the mucosal innervation and the interface between nerve fibers and follicular dendritic cells (FDC), two dramatic influences on neuroinvasion. Bovine IPP were studied at three ages, viz., newborn calves, calves less than 12 months old, and bovines older than 24 months, and the parameters obtained were compared with those of JPP. No differences in innervation patterns between IPP and JPP were found. The major difference observed was that, in calves of less than 12 months, IPP were the major mucosal-associated lymphoid organ that possessed a large number of follicles with extended FDC networks. Using a panel of antibodies, we showed that PP in 24-month-old bovines were highly innervated at various strategic sites assumed to be involved in the invasion and replication of the BSE pathogen: the suprafollicular dome, T cell area, and germinal centers. In PP in calves of less than 12 months old, no nerve fibers positive for the neurofilament markers NF-L (70 kDa) and NF-H (200 kDa) were observed in contact with FDC. Thus, in view of the proportion of these protein subunits present in neurofilaments, the innervation of the germinal centers can be said to be an age-dependent dynamic process. This variation in innervation might influence the path of neuroinvasion and, thus, the susceptibility of bovines to the BSE agent. PMID- 17406904 TI - Photosynthesis, chlorophyll fluorescence and spectral reflectance in Sphagnum moss at varying water contents. AB - Moss samples from the Fluxnet-Canada western peatland flux station in the Boreal Region of Alberta were measured in the laboratory to obtain the net photosynthesis rate and chlorophyll fluorescence of the moss under controlled environmental conditions, including the regulation of moss water content, simultaneously with measurements of moss spectral reflectance. One objective was to test whether the photochemical reflectance index (PRI) detected changes in moss photosynthetic light-use efficiency that were consistent with short-term (minutes to hours) changes in xanthophyll cycle pigments and associated changes in non-photochemical quenching (NPQ), as recorded by chlorophyll fluorescence. The rate of net photosynthesis was strongly inhibited by water content at values exceeding approximately 9 (fresh weight/dry weight) and declined as the water content fell below values of approximately 8. Chlorophyll fluorescence measurements of maximum photosystem II efficiency generally remained high until the water content was reduced from the maximum of about 20 to values of approximately 10-11, and then declined with further reductions in moss water content. A significant linear decline in NPQ was observed as moss water content was reduced from maximum to low water content values. There was a strong negative correlation between changes in NPQ and PRI. These data suggest that PRI measurements are a good proxy for short-term shifts in photosynthetic activity in Sphagnum moss. A second objective was to test how accurately the water band index (WBI, ratio of reflectance at 900 and 970 nm) recorded changes in moss water content during controlled laboratory studies. Strong linear relationships occurred between changes in moss water content and the WBI, although the slopes of the linear relationships were significantly different among sample replicates. Therefore, WBI appeared to be a useful tool to determine sample-specific water content without destructive measurements. PMID- 17406905 TI - Effects of nutritional restriction on nitrogen and carbon stable isotopes in growing seabirds. AB - When using stable isotopes as dietary tracers it is essential to consider effects of nutritional state on isotopic fractionation. While starvation is known to induce enrichment of (15)N in body tissues, effects of moderate food restriction on isotope signatures have rarely been tested. We conducted two experiments to investigate effects of a 50-55% reduction in food intake on delta(15)N and delta(13)C values in blood cells and whole blood of tufted puffin chicks, a species that exhibits a variety of adaptive responses to nutritional deficits. We found that blood from puffin chicks fed ad libitum became enriched in (15)N and (13)C compared to food-restricted chicks. Our results show that (15)N enrichment is not always associated with food deprivation and argue effects of growth on diet-tissue fractionation of nitrogen stable isotopes (Delta(15)N) need to be considered in stable isotope studies. The decrease in delta(13)C of whole blood and blood cells in restricted birds is likely due to incorporation of carbon from (13)C-depleted lipids into proteins. Effects of nutritional restriction on delta(15)N and delta(13)C values were relatively small in both experiments (delta(15)N: 0.77 and 0.41 per thousand, delta(13)C: 0.20 and 0.25 per thousand) compared to effects of ecological processes, indicating physiological effects do not preclude the use of carbon and nitrogen stable isotopes in studies of seabird ecology. Nevertheless, our results demonstrate that physiological processes affect nitrogen and carbon stable isotopes in growing birds and we caution isotope ecologists to consider these effects to avoid drawing spurious conclusions. PMID- 17406906 TI - Stage-dependent patterns of drought tolerance and gas exchange vary between sexes in the alpine willow, Salix glauca. AB - Females and males of sexually dimorphic species have distinct resource demands due to differential allocation to reproduction. Sexual allocation theory predicts that functional traits will diverge between sexes to support these demands. However, such dimorphism may be masked by the impact of current reproduction on source-sink interactions between vegetative and reproductive organs. We ask whether natural selection has led to genetic dimorphism in homologous physiological traits between sexes of the dioecious willow shrub, Salix glauca. In a common garden experiment we compared physiological responses to drought stress by male and female ramets in the absence of confounding demands from reproductive structures. Ramets experienced similar pre-dawn leaf water status (Psi(l)) as parental genets in flower within the natural population, indicating that experimental dry-down mirrored environmental conditions in nature. Male and female ramets achieved similar instantaneous water use efficiency, based on the ratio of carbon gain to water loss, under wet and dry conditions. However, female ramets experienced greater water stress (i.e., more negative Psi(l)) than males under dry conditions. Lower Psi(l) for female ramets may partly reflect the maintenance of conductance under drought; males, in contrast, maintain Psi(l) under drought by reducing conductance. Differences between sexes in terms of conductance and leaf water status of the vegetative ramets were absent in a concomitant comparison of parental flowering plants. Our results show (1) genetic divergence in physiology between sexes of S. glauca occurs in the absence of gender-specific reproductive sinks, (2) males are the more physiologically plastic sex with respect to water use, and (3) paradoxically, divergence in water relations between sexes is not detectable at sexual maturity under natural conditions. PMID- 17406907 TI - Mycorrhizal status of Eucalyptus plantations in south China and implications for management. AB - The aim of this study is to assess the mycorrhizal status of Eucalyptus plantations in south China and to determine the need for inoculation. In four provinces in south China, 155 plantations were sampled for sporocarps of ectomycorrhizal (ECM) fungi, spores of arbuscular mycorrhizal (AM) fungi, and mycorrhizas over 2 years. This study revealed a low above-ground diversity of ECM fungi consisting of 15 taxa fruiting beneath Eucalyptus plantations. The most common ECM genera were Scleroderma and Pisolithus, but they were infrequent. A total of 21 AM fungi, mostly Glomus species, were recognized from spores collected from eucalypt plantations. Four Glomus species were frequently present in soils, but spore density and relative abundance of AM fungi were generally low. Eucalypt roots from all plantation sites were poorly colonized by either ECM fungi or AM fungi. A bioassay with E. urophylla as a bait host, using soils collected from 11 eucalypt plantations, confirmed low levels of inoculum of both ECM and AM fungi in field soil. This is the first integrated study on the mycorrhizal status of eucalypt plantations in China. Findings from this research can be used to encourage adoption of mycorrhizal technology by eucalypt nurseries in the region. The potential of using spores of compatible ECM fungi or collections for forest nurseries is discussed. PMID- 17406908 TI - Surgical treatment strategies and outcome in patients with breast cancer metastatic to the spine: a review of 87 patients. AB - Aggressive surgical management of spinal metastatic disease can provide improvement of neurological function and significant pain relief. However, there is limited literature analyzing such management as is pertains to individual histopathology of the primary tumor, which may be linked to overall prognosis for the patient. In this study, clinical outcomes were reviewed for patients undergoing spinal surgery for metastatic breast cancer. Respective review was done to identify all patients with breast cancer over an eight-year period at a major cancer center and then to select those with symptomatic spinal metastatic disease who underwent spinal surgery. Pre- and postoperative pain levels (visual analog scale [VAS]), analgesic medication usage, and modified Frankel grade scores were compared on all patients who underwent surgery. Univariate and multivariate analyses were used to assess risks for complications. A total of 16,977 patients were diagnosed with breast cancer, and 479 patients (2.8%) were diagnosed with spinal metastases from breast cancer. Of these patients, 87 patients (18%) underwent 125 spinal surgeries. Of the 76 patients (87%) who were ambulatory preoperatively, the majority (98%) were still ambulatory. Of the 11 patients (13%) who were nonambulatory preoperatively, four patients were alive at 3 months postoperatively, three of which (75%) regained ambulation. The preoperative median VAS of six was significantly reduced to a median score of two at the time of discharge and at 3, 6, and 12 months postoperatively (P < 0.001 for all time points). A total of 39% of patients experienced complications; 87% were early (within 30 days of surgery), and 13% were late. Early major surgical complications were significantly greater when five or more levels were instrumented. In patients with spinal metastases specifically from breast cancer, aggressive surgical management provides significant pain relief and preservation or improvement of neurological function with an acceptably low rate of complications. PMID- 17406909 TI - Torsion of a hyperstimulated ovary during pregnancy: a potentially difficult diagnosis. AB - Ovarian torsion in a pregnant infertility patient is an important and difficult diagnosis to make. We present two cases of torsed hyperstimulated ovaries where grey scale and Doppler sonographic evaluation were inconclusive. In one case, magnetic resonance imaging (MRI) was used to make the diagnosis. In the second case, physical findings observed during the ultrasound exam were diagnostic. We think these two cases are good teaching examples of how to approach this difficult diagnosis and emphasize that MRI and physical exam skills are useful adjuncts to ultrasound in the evaluation of a pregnant patient. PMID- 17406910 TI - CT features of descending necrotizing mediastinitis--a pictorial essay. AB - Descending necrotizing mediastinitis (DNM) is a relatively rare condition caused by downward spread of neck infections into the mediastinum. This infection previously had a much worse prognosis. In recent years, prompt computer topography (CT) diagnosis has been recommended. CT scan provides the earliest means of detecting DNM for optimal management and early surgical intervention. This paper provides an illustrated summary of our extensive clinical experience with DNM, involving 36 documented cases with CT over a 5-year period. PMID- 17406911 TI - Endovascular repair of a pseudo-aneurysm of the common femoral artery after revision total hip arthroplasty. AB - Vascular injuries with acute arterial haemorrhage are rare but the infrequency of these complications may make their diagnosis and treatment challenging for surgeons unfamiliar with their management. In the past, surgery or coil embolisation has been used to treat these lesions; we describe the case of a pseudo-aneurysm of the common femoral artery developed during a total hip arthroplasty, promptly managed with an endovascular stent graft. PMID- 17406912 TI - Choroid plexitis in a case of systemic nocardiosis. AB - Although rare, primary choroid plexitis can occur as an early presentation of a central nervous system (CNS) infection most commonly with cryptococcosis, tuberculosis, and nocardiosis. In the appropriate clinical setting, an enlarged, intensely enhancing choroid plexus should raise suspicion for choroid plexitis. It is important to recognize this entity early as aggressive diagnostic and therapeutic intervention may be necessary. We review the existing literature and present a case of infectious choroid plexitis in a patient with systemic nocardiosis; computed tomography and magnetic resonance imaging demonstrated the characteristic findings of choroid plexitis, which later developed into a parenchymal abscess. PMID- 17406913 TI - CT findings of normal and inflamed appendix in groin hernia. AB - Acute appendicitis with the vermiform appendix located in a groin hernia is a rare condition. The preoperative diagnosis is important to decrease morbidity. We describe the computed tomography imaging characteristics of three cases of normal and inflamed appendices in inguinal and femoral hernias. We provide a review of the literature and consider the implications for surgical management. PMID- 17406914 TI - Renal medullary carcinoma: unsuspected diagnosis at stone protocol CT. AB - Renal medullary carcinoma (RMC) is an aggressive neoplasm occurring almost exclusively in adolescents and young adults with sickle cell (SC) hemoglobinopathies, usually sickle cell trait (SCT) or hemoglobin SC disease. The most common presentations are hematuria and flank or abdominal pain. It is a highly malignant tumor, and responses to chemotherapy are rare and transient resulting in a dismal prognosis. A high level of suspicion is necessary when evaluating at risk patients presenting with hematuria or flank pain, as currently it appears that only early diagnosis could potentially alter the outcome of this disease. We report a case of RMC in a young male patient with SCT, who presented to the emergency department with low back pain and microscopic hematuria, clinically mimicking acute obstructing urolithiasis. Our case emphasizes the need to consider alternate diagnoses when evaluating computed tomography scans for acute flank pain. PMID- 17406915 TI - Genome-wide expression profiling in Geobacter sulfurreducens: identification of Fur and RpoS transcription regulatory sites in a relGsu mutant. AB - Rel(Gsu) is the single Geobacter sulfurreducens homolog of RelA and SpoT proteins found in many organisms. These proteins are involved in the regulation of levels of guanosine 3', 5' bispyrophosphate, ppGpp, a molecule that signals slow growth and stress response under nutrient limitation in bacteria. We used information obtained from genome-wide expression profiling of the rel(Gsu) deletion mutant to identify putative regulatory sites involved in transcription networks modulated by Rel(Gsu) or ppGpp. Differential gene expression in the rel(Gsu) deletion mutant, as compared to the wild type, was available from two growth conditions, steady state chemostat cultures and stationary phase batch cultures. Hierarchical clustering analysis of these two datasets identified several groups of operons that are likely co-regulated. Using a search for conserved motifs in the upstream regions of these co-regulated operons, we identified sequences similar to Fur- and RpoS-regulated sites. These findings suggest that Fur- and RpoS-dependent gene expression in G. sulfurreducens is affected by Rel(Gsu)-mediated signaling. PMID- 17406916 TI - Optimal biocatalyst loading in a fixed bed. AB - The optimal distribution of biocatalyst in a fixed bed operating at steady state was determined to minimize the length of the bed for a fixed conversion of 95%. The distribution in terms of the biocatalyst loading on an inert support depends upon the axial distance from the bed entrance (continuous solution) as well as a set of dimensionless parameters that reflect the bed geometry, the bulk flow, reaction kinetics and diffusional characteristics. A mathematical model of the system with the following features was used to obtain the results: (1) convective mass transfer and dispersion in the bulk phase; (2) mass transfer from the bulk phase to the surface of the catalyst particle; and (3) simultaneous diffusion and chemical reaction in the catalyst particle with Michaelis-Menton kinetics and a reliable diffusion model (Zhao and DeLancey in Biotechnol Bioeng 64:434-441, 1999, 2000). The solution to the mathematical model was obtained with Mathematica utilizing the Runge Kutta 4-5 method. The dimensionless length resulting from the continuous solution was compared with the optimal length restricted to a uniform or constant cell loading across the entire bed. The maximum difference in the dimensionless length between the continuous and uniform solutions was determined to be 6.5%. The model was applied to published conversion data for the continuous production of ethanol that included cell loading (Taylor et al. in Biotechnol Prog 15:740-751, 2002). The data indicated a minimum production cost at a catalyst loading within 10% of the optimum predicted by the mathematical model. The production rate versus cell loading in most cases displayed a sufficiently broad optimum that the same (non-optimal) rate could be obtained at a significantly smaller loading such as a rate at 80% loading being equal to the rate at 20% loading. These results are particularly important because of the renewed interest in ethanol production (Novozymes and BBI International, Fuel ethanol: a technological evolution, 2004). PMID- 17406917 TI - Prospective coding in event representation. AB - A perceived event such as a visual stimulus in the external world and a to-be produced event such as an intentional action are subserved by event representations. Event representations do not only contain information about present states but also about past and future states. Here we focus on the role of representing future states in event perception and generation (i.e., prospective coding). Relevant theoretical issues and paradigms are discussed. We suggest that the predictive power of the motor system may be exploited for prospective coding not only in producing but also in perceiving events. Predicting is more advantageous than simply reacting. Perceptual prediction allows us to select appropriate responses ahead of the realization of an (anticipated) event and therefore, it is indispensable to flexibly and timely adapt to new situations and thus, successfully interact with our physical and social environment. PMID- 17406918 TI - The symbol detachment problem. AB - In situated and embodied approaches it is commonly assumed that the dynamics of sensorimotor engagement between an adaptive agent and its environment are crucial in understanding natural cognition. This perspective permits to address the symbol grounding problem, since the aboutness of any mental state arising during agent-environment engagement is guaranteed by their continuous coupling. However, cognitive agents are also able to formulate representations that are detached from the current state of affairs, such as expectations and goals. Moreover, they can act on their representations before--or instead of--acting directly on the environment, for example building the plan of a bridge and not directly the bridge. On the basis of representations, actions such as planning, remembering or imagining are possible that are disengaged from the current sensorimotor cycle, and often functional to future-oriented conducts. A new problem thus has to be acknowledged, the symbol detachment problem: how and why do situated agents develop representations that are detached from their current sensorimotor interaction, but nevertheless preserve grounding and aboutness? How do cognitive agents progressively acquire a range of capabilities permitting them to deal not only with the current situation but also with alternative, in particular future states of affairs? How do they develop the capability of acting on their representations instead of acting directly on the world? In a theoretical and developmental perspective, we propose that anticipation plays a crucial role in the detachment process: anticipatory representations, originally detached from the sensorimotor cycle for the sake of action control, are successively exapted for bootstrapping increasingly complex cognitive capabilities. PMID- 17406919 TI - Amisulpride in the short-term treatment of depressive and physical symptoms in cancer patients during chemotherapies. AB - INTRODUCTION: Amisulpride is a substituted benzamide that, at low doses, selectively blocks D2 and D3 presynaptic dopamine receptors, enhancing dopaminergic transmission in frontal cortex and limbic areas. Many clinical studies versus placebo, tricyclic antidepressants and selective serotonin reuptake inhibitors showed amisulpride antidepressant effect, supporting its safety and rapid onset of action. In oncological population, depression is quite frequent and difficult to treat because of the particular sensitivity of cancer patients to the antidepressants' side effects. GOALS OF WORK: The aims of this study were to evaluate efficacy, safety and tolerability of low doses of amisulpride (50 mg) in oncological, depressed patients during chemotheraphy. MATERIALS AND METHODS: One hundred six consecutive cancer outpatients with depressive symptoms were treated in a prospective, intention to treat, 4-week study, and were evaluated in single-blind with Montgomery Asberg rating scale for depression (MADRS), clinical global impression (CGI) and dosage record treatment emergent symptom scale (DOTES) to assess side effects of treatment. MAIN RESULTS: After 4 weeks of treatment, scores of MADRS and CGI significantly improved (p < 0.002; p < 0.001, respectively), with a reduction of depressive symptoms concerning both emotional (such as apparent sadness, reported sadness, inner tension, etc.) and physical cluster (such as lack of appetite, reduction in weight, tiredness and insomnia) with good tolerability (only two patients dropped out). CONCLUSIONS: This study is the first trial on the use of amisulpride in a cohort of oncological, depressed patients during chemotherapy. Amisulpride demonstrated high efficacy and safety. Controlled studies are needed to confirm these preliminary data. PMID- 17406921 TI - [Foudroyant course of a therapy resistent Wegener's granulomatosis with negative c-ANCA]. AB - BACKGROUND: Wegener's Granulomatosis is a rare systemic disease characterized by the triad of necrotizing granulomas of blood vessels, upper respiratory tract, and kidneys. Diagnosis depends on clinical symptoms (ACR-criteria), detection of serum circulating antineutrophil cytoplasmic antibodies (c-ANCA) and bioptical histopathology. CASE-REPORT: A 34-year-old patient presented with intraoral ulcerating, necrotizing inflammations of the palate and the gingiva. Chest radiograph (nodules, infiltrates), urin analysis (red blood cell casts) being inconspicuous Wegener's granulomatosis was diagnosed by histopathology of an intraoral incisional biopsy. Conventional therapy with corticosteroids and cyclophosphamide relapsed while the disease was taking a progressive course (pulmonary infiltrates, necrotizing vasculitides all over the common integument). The patient died of cardiac decompensation three months after diagnosis. c-ANCAs were not present at any time of disease. CONCLUSIONS: 1. Symptoms of the head and neck are manifest in up to 95% of Wegener's granulomatosis. So knowledge of the disease is important for dentists and oral and maxillofacial surgeons in spite of its low prevalence. 2. c-ANCA-titers do not reflect disease activity. Negative c ANCAs do not exclude Wegener's granulomatosis. 3. Wegener's granulomatosis can take a progressive, therapy resistent course. PMID- 17406922 TI - [Surgical management of obstructive sleep apnea]. AB - INTRODUCTION: Obstructive sleep apnea (OSA) is a common disorder in adults, however it also occurs in neonates with severe mandibular hypoplasia. The clinical significance of OSA results from hypoxemia and sleep fragmentation due to a collapse of the upper airway. Recently OSA has received much attention due to its potentially serious consequences on cardiac function. Besides daytime sleepiness and fatigue life expectancy is significantly shortened by a higher risk for apoplectic insults and heart failures. This paper reviews common surgical procedures with an emphasis on jaw advancement procedures. MATERIAL AND METHODS: In six adults (mean age 46,9 ys; 2 female, 4 male; BMI averaged 28kg/m(2)) suffering from OSA with a history of a long-lasting unsuccessful continuous positive airway pressure (CPAP) therapy a bimaxillary advancement was performed. The respiratory disturbance was preoperatively evaluated by polysomnography; daytime sleepiness was recorded according to the 'Epworth Sleepiness Scale'. Controls were performed six months after therapy. All patients had a regular intermaxillary relation. Bimaxillary advancement was planned and performed according to the principles of cranio-maxillofacial corrective surgery. To widen the posterior airway space much attention was focused on the advancement of the mandible by 10mm. RESULTS: The apnea/hypopnea index decreased from preoperatively 37/h to 4/h after treatment. The minimal peripheral oxygen saturation increased from 78% to 89%, whereas the average oxygen saturation raised from 92 to 95%. The arousal index decreased from 24/h to 9/h. Periods of snoring were reduced from 29,5% to 6,9%. According to the 'Epworth Sleepiness Scale' the patients' estimation improved from a level of 11 to 4. Sleep had a recuperating effect. All surgical procedures were uneventful. CONCLUSION: Standard treatment of OSA is the CPAP-therapy. In some patients, however, CPAP therapy cannot be applied, fails or is not well accepted. A great variety of surgical procedures for the treatment of OSA have been described. Bimaxillary advancement in adults and osteodistraction in neonates are safe and successful surgical techniques for an immediate improvement in OSA. Osteodistraction treatment can help to avoid tracheotomy in neonates. PMID- 17406924 TI - Leucine and calcium regulate fat metabolism and energy partitioning in murine adipocytes and muscle cells. AB - Dietary calcium modulation of adiposity is mediated, in part, by suppression of calcitriol, while the additional effect of dairy products is mediated by additional components; these include the high concentration of leucine, a key factor in the regulation of muscle protein turnover. We investigated the effect of leucine, calcitriol and calcium on energy metabolism in murine adipocytes and muscle cells and on energy partitioning between adipocytes and skeletal muscle. Leucine induced a marked increase in fatty acid oxidation in C2C12 muscle cells (P<0.001) and decreased FAS expression by 66% (P<0.001) in 3T3-L1 adipocytes. Calcitriol decreased muscle cell fatty acid oxidation by 37% (P<0.001) and increased adipocyte FAS gene expression by threefold (P<0.05); these effects were partially reversed by either leucine or calcium channel antagonism with nifedipine. Co-culture of muscle cells with adipocytes or incubation with 48-h adipocyte conditioned medium decreased muscle fatty acid oxidation by 62% (P<0.001), but treating adipocytes with leucine and/or nifedipine attenuated this effect. Leucine, nifedipine and calcitriol also modulated adiponectin production and thereby exerted additional indirect effects on fatty acid oxidation in C2C12 myotubes. Adiponectin increased IL-15 and IL-6 release by myotubes and partially reversed the inhibitory effects of calcitriol. Comparable effects of leucine, calcitriol and adiponectin were found in myotubes treated with conditioned medium derived from adipocytes or co-cultured with adipocytes. These data suggest that leucine and nifedipine promote energy partitioning from adipocytes to muscle cells, resulting in decreased energy storage in adipocytes and increasing fatty acid utilization in muscle. PMID- 17406925 TI - Rapid desensitization of lipolysis in the visceral and subcutaneous adipocytes of rats. AB - In adipocytes, short and long term stimulation of beta adrenergic receptors (beta AR) induces the desensitization to catecholamines, leading to a decrease in the intracellular accumulation of cAMP, but the roles played by this in lipolysis is not clear. In this study, we assessed the catecholamine-induced desensitization of lipolysis and compared this in adipocytes isolated from visceral and subcutaneous fat tissues of rats. When adipocytes were pretreated with isoproterenol (ISO), the norepinephrine (NE)-induced lipolysis was significantly reduced dose- and time-dependently. A similar reduction of the lipolytic response was also found in NE-, dobutamine-, terbutaline- or BRL37344-induced lipolysis. The ISO- and each beta AR agonist-induced lipolysis in the visceral fat was not only higher than in the subcutaneous fat, but also markedly reduced by ISO- or NE pretreatment. These results showed that short-term treatment of three subtypes of beta AR by each agonist induces a rapid reduction in the lipolytic response to beta AR stimulation. This suggests some common mechanism for the rapid desensitization of beta AR-agonist-induced lipolysis, in contrast with previous reports on the characteristics of beta AR subtypes. In addition, the regional difference of adipose tissue not only in inducing lipolysis but also in rapid desensitization was also apparent. PMID- 17406926 TI - Effect of interesterification of palmitic acid-rich triacylglycerol on postprandial lipid and factor VII response. AB - The process of interesterification results in changes in triacylglycerol (TAG) structure and is used to increase the melting point of dietary fats. The acute health effects of this process on palmitic acid-rich fats are uncertain with regard to postprandial lipemia, insulin and factor VII activated (FVIIa) concentrations. Two randomized crossover trials in healthy male subjects compared the effects of meals containing 50 g fat [interesterified palm oil (IPO) versus native palm oil (NPO); n=20, and IPO versus high-oleic sunflower oil (HOS); n=18], on postprandial changes in lipids, glucose, insulin, chylomicron composition and FVIIa. Compared with NPO, IPO decreased postprandial TAG and insulin concentrations. Both NPO and IPO increased FVIIa concentrations postprandially; mean increases at 6 h were 21 and 19%, respectively. Compared with HOS, IPO decreased postprandial TAG (47% lower incremental area under the curve) and reduced the postprandial increase in FVIIa concentration by 64% at 6 h; no significant differences in hepatic and total lipase activities or insulin concentrations were noted. All three test meals increased postprandial leukocyte counts (average 26% at 6 h). The fatty acid composition of the chylomicron TAG was similar to the test fats following all test meals. It is concluded that interesterification of palm oil does not result in adverse changes in postprandial lipids, insulin or FVIIa compared to high oleate and native palm oils. PMID- 17406927 TI - Stearidonic acid increases the red blood cell and heart eicosapentaenoic acid content in dogs. AB - Plant sources of omega-3 fatty acids (FA) are needed that can materially raise tissue levels of long-chain omega-3 FA [i.e., eicosapentaenoic acid (EPA; 20:5n 3) and docosahexaenoic acid (DHA; 20:6n-3)]. Stearidonic acid (SDA; 18:4n-3) is the delta-6 desaturase product of alpha-linolenic acid (ALA; 18:3n-3), and when fed to humans, increases red blood cell (RBC) content of EPA to a greater extent than does ALA. This study was undertaken to determine the dose-dependence and time course of the increase in the EPA and DHA content of the heart and RBC in dogs. Adult male Beagles were fed 21, 64, or 193 mg/kg of SDA in in their food daily for up to 12 weeks. Positive and negative controls were given EPA (43 mg/kg) or high oleic acid sunflower oil, respectively. The baseline EPA content of RBC was 0.38 +/- 0.03% which increased (P<0.01) in a dose-dependent manner, with the high dose of SDA and EPA achieving levels of 1.33 +/- 0.26 and 1.55. +/- 0.28%, respectively. In the heart, the content of EPA rose from 0.06 +/- 0.01 to 1.24 +/- 0.22% in the EPA group and to 0.81 +/- 0.32% in the high SDA group (both P<0.01). In both tissues, DHA did not change. Compared to dietary EPA, SDA was 20 23% as efficient in raising tissue EPA levels. In conclusion, SDA supplementation increased the EPA content of RBC and heart and may have utility as a plant-based source of omega-3 FA. PMID- 17406928 TI - Longitudinal assessment of erythrocyte fatty acid composition throughout pregnancy and post partum. AB - Transfer of fatty acids from mother to fetus during pregnancy is a requirement for optimal fetal growth. We report a longitudinal study of full maternal erythrocyte fatty acid profile assessed at each trimester of pregnancy [mean 12.5 (range 8-14), 26.1 (24-28) and 35.5 (33-38) weeks' gestation] and in the post partum period [18.1 (12-26) weeks]. The study recruited healthy women (n=47) from routine antenatal clinics at the Princess Royal Maternity Unit, Glasgow, Scotland. There were increases in 16:1n7 (22%, p=0.0005), 24:1n9 (13%, p=0.0032), 22:5n6 (25%, p=0.0003), 18:3n3 (41%, p=0.0007) and 22:6n3 (20%, p=0.0005) concentrations during pregnancy. The greatest increases took place between gestations at sampling of 12.5 and 26.1 weeks. The change in 16:1n7 concentration between gestations at sampling of 12.5 and 35.3 weeks was negatively associated with maternal booking body mass index (r=-0.40, p=0.006). The change in 22:6n3 concentration was correlated with the change in 24:1n9 (r=0.70, p<0.001). In samples taken four months post partum, 14:0 concentration was lower (29%, p=0.0002) and 24:0 concentration (15%, p=0.0009) and n6/n3 ratio (11%, p=0.0019) were higher than at a gestation at sampling of 12.5 weeks. In conclusion, several fatty acids are specifically mobilised during pregnancy. The correlation between maternal 22:6n3 and 24:1n9 suggests that mobilisation of these fatty acids may be coordinated. The inverse relationship between 16:1n7 and maternal central obesity warrants further investigation. PMID- 17406930 TI - Dynamic features of the rumen metabolism of linoleic acid, linolenic acid and linseed oil measured in vitro. AB - The lipid quality of ruminant products is largely determined by the extent of rumen microbial biohydrogenation (BH) of polyunsaturated fatty acids (FAs) and the substances formed thereby. In vitro batch incubations with mixed rumen bacteria were tracked over 24 h to characterize the profiles and kinetics of the BH products from three lipid sources: pure linoleic acid (c9,c12-18:2), pure linolenic acid (c9,c12,c15-18:3) and linseed oil (mainly c9,c12,c15-18:3 in triacylglycerols). After 24 h of incubation biohydrogenation was more complete for c9,c12-18:2, which gave mainly stearic acid (18:0), than for c9,c12,c15-18:3, which yielded mainly trans-18:1 FAs. This suggests inhibition of the final BH step (18:1 to 18:0). Incubations of c9,c12-18:2 resulted in high levels of carbon 10- and 12-desaturated 18:1, t10,c12- and c9,t11-CLAs. Incubations of c9,c12,c15 18:3 resulted in high levels of t11-18:1, carbon 13- and 15-desaturated 18:1 as well as t11,c15-18:2 and 11,13-CLAs. A comparative study of linolenic acid and linseed oil kinetics revealed that the BH process was not significantly slowed by the esterification of polyunsaturated FAs, but may have been limited by the isomerization step in which the cis12 double bond goes to the trans11 position. The disappearance rates of c9,c12-18:2 and c9,c12,c15-18:3 ranged from 23.6 to 44.6%/h. The wide variety of BH intermediates found here underlines the large number of possible BH pathways. These data help provide a basis for dynamic approaches to BH processes. PMID- 17406929 TI - N-3 polyunsaturated fatty acids and atopy in Korean preschoolers. AB - Atopy is a growing problem for Korean children. Since eicosapentaenoic acid is a precursor of less active inflammatory eicosanoids, n-3 polyunsaturated fatty acids (PUFA) may have a protective effect on atopy. This study was undertaken to determine whether n-3 PUFA in red blood cells (RBC) is lower in atopic than in non-atopic preschoolers. Three hundred and eight Korean children aged 4-6 years were enrolled. Total RBC fatty acid composition was measured by gas chromatography. The prevalence of atopic dermatitis, allergic rhinitis, or asthma was 29%. Total RBC n-3 PUFA were lower in preschoolers with atopy than controls (9.8 +/- 1.2 vs. 11.4 +/- 1.6%; P<0.05), while n-6 PUFA (33.0 +/- 1.4 vs. 32.2 +/ 1.0%; P<0.05) and n-6/n-3 PUFA ratio (3.4 +/- 0.6 vs. 2.8 +/- 0.5; P<0.05) were greater. The following factors were also associated with an increase in atopy: higher saturated fatty acids (39.6 +/- 1.4 vs. 40.6 +/- 1.9; P<0.05) and arachidonic acid (15.3 +/- 1.6 vs. 16.0 +/- 2.9; P<0.05), and lower total PUFA (43.8 +/- 0.7 vs. 42.8 +/- 1.4; P<0.05) and omega-3 index (EPA + DHA; 9.1 +/- 0.8 vs. 7.8 +/- 0.5; P<0.05) in RBC. Maternal history of atopy was a significant (P<0.05) risk factor, while lactation was not. The results suggest that a reduced content of n-3 PUFA in the RBC membrane could play a role in early children atopy. PMID- 17406931 TI - Total lipids of Sarda sheep meat that include the fatty acid and alkenyl composition and the CLA and trans-18:1 isomers. AB - The total lipids of the longissimus dorsi muscle were analyzed from commercial adult Sarda sheep in Sardina taken from local abattoirs, and in the subsequent year from three local farms in the Sassari region that provided some information on the amount and type of supplements fed to the pasture-fed sheep. The complete lipid analysis of sheep meat included the fatty acids from O-acyl and N-acyl lipids, including the trans- and conjugated linoleic acid (CLA) isomers and the alk-1-enyl ethers from the plasmalogenic lipids. This analysis required the use of a combination of acid- and base-catalyzed methylation procedures, the former to quantitate the O-acyl, N-acyl and alkenyl ethers, and the latter to determine the content of CLA isomers and their metabolites. A combination of gas chromatographic and silver-ion separation techniques was necessary to quantitate all of the meat lipid constituents, which included a prior separation of the trans-octadecenoic acids (18:1) and a separation of fatty acid methyl esters and the dimethylacetals (DMAs) from the acyl and alk-1-enyl ethers, respectively. The alk-1-enyl moieties of the DMAs were analyzed as their stable cyclic acetals. In general, about half of the meat lipids were triacylglycerols, even though excess fat was trimmed from the meat. The higher fat content in the meat appears to be related to the older age of these animals. The variation in the trans-18:1 and CLA isomer profiles of the Sarda sheep obtained from the abattoirs was much greater than in the profiles from the sheep from the three selected farms. Higher levels of 10t-18:1, 7t9c-18:2, 9t11c-18:2 and 10t12c-18:2 were observed in the commercial sheep meat, which reflected the poorer quality diets of these sheep compared to those from the three farms, which consistently showed higher levels of 11t-18:1, 9c11t-18:2 and 11t13c-18:2. In the second study, sheep were provided with supplements during the spring and summer grazing season, which contributed to higher levels of 11t-18:1 and 9c11t-18:2. The farm that provided a small amount of supplements during the spring had the better lipid profile at both time periods. The polyunsaturated fatty acid (PUFA) content was higher in the meat from Sarda sheep from the three farms than in the meat from those sheep obtained from commercial slaughter operations. The plasmalogenic lipid content ranged from 2 to 3% of total lipids, the alk-1-enyl ethers consisted mainly of saturated and monounsaturated moieties, and the trans-18:1 profile was similar to that of the FA. The n-6 (6-8%) and n-3 PUFA (2-3%) contents, the n-6/n-3 ratio (3:1), as well as the saturated fatty acid (SFA) content (42-45%) and the SFA to PUFA ratio (4:1 to 5:1) of the Sarda sheep from the three farms were comparable to sheep meat lipids found in similar commercial operations in Europe. Inclusion of small amounts of supplements for the grazing Sarda sheep resulted in improved quality of sheep meat lipids. PMID- 17406932 TI - Comparison of the structures of triacylglycerols from native and transgenic medium-chain fatty acid-enriched rape seed oil by liquid chromatography- atmospheric pressure chemical ionization ion-trap mass spectrometry (LC-APCI ITMS). AB - The sn position of fatty acids in seed oil lipids affects physiological function in pharmaceutical and dietary applications. In this study the composition of acyl chain substituents in the sn positions of glycerol backbones in triacylglycerols (TAG) have been compared. TAG from native and transgenic medium-chain fatty acid enriched rape seed oil were analyzed by reversed-phase high performance liquid chromatography coupled with online atmospheric-pressure chemical ionization ion trap mass spectrometry. The transformation of summer rape with thioesterase and 3 ketoacyl-[ACP]-synthase genes of Cuphea lanceolata led to increased expression of 1.5% (w/w) caprylic acid (8:0), 6.7% (w/w) capric acid (10:0), 0.9% (w/w) lauric acid (12:0), and 0.2% (w/w) myristic acid (14:0). In contrast, linoleic (18:2n6) and alpha-linolenic acid (18:3n3) levels decreased compared with the original seed oil. The TAG sn position distribution of fatty acids was also modified. The original oil included eleven unique TAG species whereas the transgenic oil contained sixty. Twenty species were common to both oils. The transgenic oil included trioctadecenoyl-glycerol (18:1/18:1/18:1) and trioctadecatrienoyl glycerol (18:3/18:3/18:3) whereas the native oil included only the latter. The transgenic TAG were dominated by combinations of caprylic, capric, lauric, myrisitic, palmitic (16:0), stearic (18:0), oleic (18:1n9), linoleic, arachidic (20:0), behenic (22:0), and lignoceric acids (24:0), which accounted for 52% of the total fat. In the original TAG palmitic, stearic, oleic, and linoleic acids accounted for 50% of the total fat. Medium-chain triacylglycerols with capric and lauric acids combined with stearic, oleic, linoleic, alpha-linolenic, arachidic, and gondoic acids (20:1n9) accounted for 25% of the transgenic oil. The medium chain fatty acids were mainly integrated into the sn-1/3 position combined with the essential linoleic and alpha-linolenic acids at the sn-2 position. Eight species contained caprylic, capric, and lauric acids in the sn-2 position. The appearance of new TAG in the transgenic oil illustrates the extensive effect of genetic modification on fat metabolism by transformed plants and offers interesting possibilities for improved enteral applications. PMID- 17406933 TI - Two new monogalactosyl diacylglycerols from brown alga Sargassum thunbergii. AB - Two new monogalactosyl diacylglycerols (MGDGs) along with two known glycolipids were isolated from the moderate polar fraction of the methanolic extract of the brown alga Sargassum thunbergii by using reversed silica flash chromatography. Two new MGDGs were identified as (2S)-1-O-(5Z,8Z,11Z,14Z,17Z-eicosapentaenoyl)-2 O-(9Z,12Z,15Z-octadecatrienoyl)-3-O-beta-D-galactopyranosyl-sn-glycerol (1) and (2S)-1-O-(9Z,12Z,15Z-octadecatrienoyl)-2-O-(6Z,9Z,12Z,15Z-octadecatetraenoyl)-3-O beta-D-galactopyranosyl-sn-glycerol (2) by FAB tandem mass spectrometry, NMR techniques, and specific enzyme-catalyzed hydrolysis of the sn-1 fatty acyl linkage. The regiochemical attachment of the acyl chains in the glycerol moiety was established by 2D NMR correlations and confirmed by enzymatic hydrolysis. PMID- 17406934 TI - Pomegranate juice may be a potential addition to anti-Behcet armamentarium: a hypothesis. AB - Behcet's disease (BD) is a systemic vasculitis. We speculate that local or systemic administration of the pomegranate juice may be useful in the treatment of BD. Immunomodulatory, antioxidants and antibacterial characteristics of pomegranate juice may rationalize this hypothesis. PMID- 17406938 TI - Aesthetic surgery and its future. PMID- 17406939 TI - Shortscar face-lift with the RoundBlock SMAS treatment: a younger face for all. AB - BACKGROUND: Performing a face-lift for smokers, bald people, and those looking for a quick recovery is always a challenge. The submucosal aponeurotic system (SMAS) plication technique with limited undermining provides great suspension and has proved to be a safe and fast way to perform a face-lift surgery. METHODS: The procedure is a suspension technique for face-lifting based on a purse-string suture around the ear that allows all the soft tissues of the face and neck to be elevated and anchored in the strong structures of the cranial muscles and aponeurosis. This allows a harmonic volumetric repositioning and results in almost no dead space because the circular plication diminishes the surgical undermining when stretched. A total of 225 face-lifts were performed. RESULTS: Because this procedure offers quick recovery and long-lasting results with minimal complications, patients become fearless about face-lift surgery. CONCLUSIONS: The RoundBlock SMAS treatment is a suspension method for face-lifts that is safe and fast, results in reduced scars and no dead space, requires no drains, and has a quick recovery time. PMID- 17406940 TI - Investigation of TXNIP (thioredoxin-interacting protein) and TRX (thioredoxin) genes for growth-related traits in pigs. AB - It is well known that TRX and its endogenous inhibitor TXNIP help sustain the cellular reduction/oxidation balance in response to various stresses and both play a crucial role in cell proliferation and growth. Five SNPs were found in TXNIP and these allowed us to map it by linkage to SSC4. Three of the SNPs were used for association analyses in three commercial pig populations (Duroc, Hampshire, and synthetic line) with more than 1200 animals. Both the single marker and haplotype analyses revealed significant effects of TXNIP on hot carcass weight, test daily gain, and lifetime daily gain. TRX was mapped on SSC1 but no significant associations with growth-related traits were found in the synthetic pig line in which the SNP was informative. The expression levels of TXNIP and TRX were then detected in two groups (fast growth and slow growth, respectively) with different genetic backgrounds for growth. Compared with the slow-growth group, TXNIP expression was significantly lower in the fast-growth group, whereas a marked increase in TRX expression was observed in fast-growth group. Our findings suggest that TXNIP has effects on growth-related traits in pigs and further investigations will be necessary to elucidate the underlying mechanisms involved. PMID- 17406941 TI - Association of HLA class II genes with idiopathic pulmonary arterial hypertension in Koreans. AB - The aim of this study was to compare the frequency of the HLA-DRB1 and HLA-DQB1 alleles in Korean patients with idiopathic pulmonary arterial hypertension (IPAH) and in normal controls and to determine any association that may exist between clinical characteristics of IPAH and specific HLA alleles. IPAH patients seen between October 1998 and September 2001 were retrospectively assessed, and 19 patients and 193 controls were HLA typed at the HLA-DRB1 and DQB1 loci. Clinical characteristics and hemodynamic parameters were reviewed. The patients with IPAH had a significantly higher frequency of the HLA-DRB1*0406 allele (18% vs. 6%, p = 0.004) and the HLA-DQB1*0302 allele (24% vs. 12%, p = 0.034), as well as a significantly higher frequency of haplotype DRB1*0406-DQB1*0302 (p = 0.0006). All 6 patients with haplotype DRB1*0406-DQB1*0302 (H+ group) were women, compared with 8 of the 13 patients lacking the DRB1*0406-DQB1*0302 haplotype (H- group), but without statistical significance. Three of 19 patients showed a positive short-term hemodynamic response to NO inhalation, all 3 of whom had the DRB1*0406 DQB1*0302 haplotype. There were no other significant differences in clinical characteristics and hemodynamic parameters between the H+ and H- groups. We conclude from this study that the HLA-DRB1*0406-DQB1*0302 haplotype is associated with IPAH in Korean patients. These results suggest that certain clinical characteristics of IPAH may be controlled in part by patients' HLA alleles. PMID- 17406943 TI - Sjogren's syndrome with multiple bullae caused by lymphocytic interstitial pneumonia. PMID- 17406942 TI - Inoculation route-dependent and allergen-specific suppressive effects of bacille Calmette-Guerin vaccination on asthmatic reactions in BALB/c mice. AB - Intranasal bacille Calmette-Guerin (BCG) vaccination causes greater suppression of ovalbumin-induced airway eosinophilia in mice than does subcutaneous vaccination. Coadministration of ovalbumin with interleukin (IL)-18 induces an ovalbumin-specific Th1 immune reaction. The purpose of this study was to examine whether the suppressive effect of BCG is dependent on the inoculation method, using various murine asthma models. Female BALB/c mice (n = 7 per group) were immunized with BCG subcutaneously or intranasally, then sensitized with ovalbumin or Dermatophagoides farinae either immediately or one week later. After provocation with one of the allergens, the mice were tested by methacholine bronchial challenge, and analyses of the inflammatory cell numbers in the airways and cytokine levels in the supernatant of concanavalin A-stimulated splenocytes were conducted. Overall, the airway responses to the allergens were significantly lower and the interferon (IFN)-gamma level was significantly higher in BCG treated mice than in untreated mice, and the number of airway eosinophils was significantly related to the IFN-gamma/IL-5 ratio (r = -0.444, p < 0.001). Subcutaneous BCG inoculation tended to have a greater suppressive effect on the development of airway hyperresponsiveness and eosinophilia than did intranasal inoculation. Concurrent BCG vaccination and D. farinae sensitization one week before ovalbumin sensitization tended to have a greater suppressive effect on airway responsiveness to methacholine induced by D. farinae aerosols than did that induced by ovalbumin aerosols. Subcutaneous BCG inoculation suppressed asthmatic reactions more remarkably than did intranasal inoculation, and concurrent BCG vaccination and allergen sensitization induced allergen-specific suppression of asthmatic reactions. PMID- 17406944 TI - Determination of the necessary total protein substitution requirements in patients with advanced stage ovarian cancer and ascites, undergoing debulking surgery. Correlation with plasma proteins. AB - BACKGROUND: Ascites is common in patients with advanced ovarian carcinoma (AOC). Its drainage during surgery affects plasma proteins. We sought to correlate the volume of ascites with the oncological parameters of the disease, calculate its drainage effect on plasma proteins, and determine the necessary substitution requirements and a clinical way of achieving that. PATIENTS AND METHODS: We evaluated 138 patients with AOC and ascites who underwent primary cytoreductive surgery. Intraoperatively found ascites and its postoperative production were evaluated. Its drainage effect on plasma proteins and the substitution requirements were determined using a mathematic formula. Human albumin(HA) and fresh frozen plasma (FFP) were used to cover these requirements. RESULTS: The intraoperative ascites was found to correlate only with the stage of the disease, while its postoperative production correlated with the residual disease. Optimally debulked patients had a mean ascites production of 128 mL on postoperative day 1 compared with 668 mL of the suboptimally debulked. This production required 3 and 5-7 days, respectively, to drop <50 mL. Plasma proteins fell on their minimum level (88.9 versus 80.8%) on the second postoperative day. The protein deficit was calculated to be 379 and 691 g/day, respectively. This deficit was substituted by administering 2 HA and 2 FFP for 3-6 days. CONCLUSIONS: Ascites drainage affects the postoperative homeostasis of plasma proteins. A mean acute drop of 12-20% is monitored on postoperative day 2. This deficit can be managed with HA and FFP for a minimum of 3 days. PMID- 17406945 TI - Pelvic exenteration affords safe and durable treatment for locally advanced rectal carcinoma. AB - BACKGROUND: Treatment of locally advanced rectal carcinoma (LARC) often involves exenterative surgery, which can be associated with high perioperative morbidity and mortality. To assist in patient selection for radical surgery, we sought to determine clinicopathologic factors influencing recurrence and disease-free survival (DFS) of LARC. METHODS: Consecutive patients with LARC undergoing exenterative surgery were retrospectively identified in our institutional database. Factors evaluated included age, sex, primary versus recurrent tumors, neoadjuvant or adjuvant chemoradiotherapy, resection margin status, recurrence, time to recurrence, and survival. The primary outcome was DFS. Secondary outcomes were overall survival and perioperative morbidity. RESULTS: A total of 72 patients were identified; median age was 52 years, and median follow-up time was 30 months. The overall complication rate was 43%; rates were similar among the patients with primary (47%) or recurrent (37%) LARC. Primary or recurrent tumor status was the only factor significantly predictive of outcome after exenteration. Local recurrence rates were lower in the primary group (primary 22%, recurrent 52%, P = .05). A significant difference in 5-year DFS was found between primary and recurrent tumor (52% vs. 13%; P < .01). The median time to recurrence was longer in the patients with primary LARC (17 months vs. 8 months; P < .01). CONCLUSIONS: The complication rates for pelvic exenteration remain high, but the morbidity can typically be managed without a clinically important increase in hospitalization. In primary LARC, an aggressive surgical approach provides most patients 5-year DFS. Select patients with recurrent LARC will also benefit from pelvic exenteration. PMID- 17406946 TI - Retroperitoneal sarcomas--an SOS to colleagues in Europe. PMID- 17406947 TI - Factors influencing the volume-outcome relationship in gastrectomies: a population-based study. AB - BACKGROUND: A relationship between hospital procedural volume and patient outcomes has been observed in gastrectomies for primary gastric cancer, but modifiable factors influencing this relationship are not well elaborated. METHODS: We performed a population-based study of 1864 patients undergoing gastrectomy for primary gastric cancers at 214 hospitals. Hospitals were stratified as high-, intermediate-, or low-volume centers. Multivariate models were constructed to evaluate the effect of institutional procedural volume and other hospital- and patient-specific factors on the risk of in-hospital mortality, adverse events, and failure to rescue, defined as mortality after an adverse event. RESULTS: High-volume centers attained an in-hospital mortality rate of 1.0% and failure-to-rescue rate of .7%, both less than one-fifth of that seen at intermediate- and low-volume centers, although adverse event rates were similar across the three volume tiers. In multivariate modeling, treatment at a high-volume hospital decreased the odds of mortality (odds ratio [OR], .22; 95% confidence interval [95% CI], .05-.89), whereas treatment at an institution with a high ratio of licensed vocational nurses per bed increased the odds of mortality (OR, 1.96; 95% CI, 1.04-3.75). Being treated at a hospital with a greater than median number of critical care beds decreased odds of mortality (OR, .46; 95% CI, .25-.81) and failure to rescue (OR, .53; 95% CI, .29-.97). CONCLUSIONS: Undergoing gastrectomy at a high-volume center is associated with lower in-hospital mortality. However, improving the rates of mortality after adverse events and reevaluating nurse staffing ratios may provide avenues by which lower-volume centers can improve mortality rates. PMID- 17406948 TI - Heterogenic loss of BRCA in breast cancer: the "two-hit" hypothesis takes a hit. PMID- 17406949 TI - Differential expression and prognostic implications of the CCN family members WISP-1, WISP-2, and WISP-3 in human breast cancer. AB - BACKGROUND: The CCN family has three Wnt-inducted secreted proteins named WISP-1, WISP-2 and WISP-3. These molecules are known to play a diverse role in cells, but their role in cancer cells remains controversial. METHODS: In this study, we analyzed the expression of the three WISP molecules at the mRNA and protein levels in a cohort of 122 human breast tumors and 32 normal breast tissues, and we correlated these findings with patients' clinical outcomes. RESULTS: WISP-1 transcripts were found in lower levels in node-positive tumors compared with node negative tumors (P < .05); were lower in patients with a moderate (P = .01) and poor Nottingham Prognostic Index prognosis (P < .05) compared with good prognostic groups; were of significantly lower level in grade 3 differentiated tumors (P < .05) compared with grade 1; and were of lower levels in patients who developed metastasis and died from breast cancer-related causes (P < .05 in both comparisons). Almost the reverse was found to be true for WISP-2, which had greater levels of expression in node-positive tumors (P = .0043); higher levels in both moderate and poor prognostic groups compared with the good prognostic group (both P < .05); greater level in both grade 2 and 3 when compared with grade 1 (both P < .05); and higher levels in patients who went on to develop metastases (P < .01). WISP-3 transcript levels showed no statistically significant differences between groups. CONCLUSIONS: WISPs may play important but contrasting roles in breast cancer. WISP-1 seems to act as a tumor suppressor and WISP-2 as a factor that stimulates aggressiveness; WISP-3 has no definable beneficial or detrimental role. PMID- 17406950 TI - High serum vascular endothelial growth factor levels predict poor prognosis after radiofrequency ablation of hepatocellular carcinoma: importance of tumor biomarker in ablative therapies. AB - BACKGROUND: Radiofrequency ablation (RFA) is a recently developed treatment for hepatocellular carcinoma (HCC). Thus far, the prognostic impact of tumor biomarkers has not been evaluated in this treatment. High serum level of vascular endothelial growth factor (VEGF) has been shown to predict microscopic vascular invasion and metastasis in HCC. This study investigated the prognostic significance of pre-treatment serum VEGF level in patients with HCC undergoing RFA treatment. METHODS: Serum VEGF levels were measured using enzyme-linked immunosorbent assay in 120 patients with HCC undergoing RFA, and in 15 healthy controls. Serum VEGF levels were correlated with clinicopathological features of the HCC patients. The prognostic significance of serum VEGF levels was assessed by univariate and multivariate analyses. RESULTS: The median serum VEGF level in the HCC patients was 240 pg/mL (range 17-1162), significantly higher than that of healthy controls (p = .024). The serum VEGF levels were significantly correlated with platelet counts (r = .487, p < .001) but not other clinicopathological features. Patients with serum VEGF level > 240 pg/mL had worse overall and recurrence-free survival compared with those with serum VEGF level > 240 pg/mL (p = .005 and .002, respectively). By multivariate analysis, serum VEGF level was a significant prognostic factor of both overall and recurrence-free survival. CONCLUSIONS: High pre-treatment serum VEGF levels predict poor prognosis after RFA of HCC. This study highlights the importance of tumor biomarker as a prognostic predictor in ablative therapy for HCC, which has an intrinsic problem of unavailability of histopathological prognostic features. PMID- 17406951 TI - Patterns of first-recurrence and post-recurrence survival in patients with primary cutaneous melanoma after sentinel lymph node biopsy. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) has become well accepted in management of patients with primary cutaneous melanoma. An understanding of the pattern of recurrence after SLNB is helpful in coordinating a rational plan of follow-up in these patients. We sought to determine the site and timing of initial recurrence and post-recurrence survival after SLNB. METHODS: Stage I/II melanoma patients who underwent SLNB during 1991-2004 were identified from a prospective single-institution database. Site and date of first recurrence after SLNB were recorded. Patterns of recurrence after SLNB and post-recurrence survival were analyzed. RESULTS: One thousand and forty-six patients underwent SLNB. The sentinel lymph node (SLN) was positive in 164 patients (16%). Median follow-up was 36 months for survivors. Median and 3-year relapse-free survival for SLN-positive patients were 41 months and 56%, and for SLN-negative patients were not reached and 87%, respectively (P < .0001). Of the SLN-positive patients, 47% experienced recurrence, compared with 14% SLN-negative patients. The pattern of recurrence stratified by SLN status was similar between the two groups (P = NS). After recurrence, the site of recurrence was the only significant prognostic factor influencing survival (P < .0001). CONCLUSIONS: Although SLN-positive patients experience recurrence far earlier and more frequently than SLN-negative patients, the pattern of recurrence is similar. After recurrence, its site is the primary determinant of survival. PMID- 17406952 TI - Strategies for coping in a complex world: adherence behavior among older adults with chronic illness. AB - BACKGROUND: Increasing numbers of medicines increase nonadherence. Little is known about how older adults manage multiple medicines for multiple illnesses. OBJECTIVES: To explore how older adults with multiple illnesses make choices about medicines. DESIGN: Semistructured interviews with older adults taking several medications. Accounts of respondents' medicine-taking behavior were collected. PARTICIPANTS: Twenty community-dwelling seniors with health insurance, in Eastern Massachusetts, aged 67-90, (4-12 medicines, 3-9 comorbidities). APPROACH: Qualitative analysis using constant comparison to explain real choices made about medicines in the past ("historical") and hypothetical ("future") choices. RESULTS: Respondents reported both past ("historical") choices and hypothetical ("future") choices between medicines. Although people discussed effectiveness and future risk of the disease when prompted to prioritize their medicines (future choices), key factors leading to nonadherence (historical choices) were costs and side effects. Specific choices were generally dominated by 1 factor, and respondents rarely reported making explicit trade-offs between different factors. Factors affecting 1 choice were not necessarily the same as those affecting another choice in the same person. There was no evidence of "adherent" personalities. CONCLUSION: Prescribing a new medicine, a change in provider or copayment can provoke new choices about both new and existing medications in older adults with multiple morbidities. PMID- 17406953 TI - Effects of anticholinergics and beta-agonists in COPD. PMID- 17406955 TI - Visits for depression by elderly patients. PMID- 17406957 TI - Obesity among veterans. PMID- 17406959 TI - Patterns of communication through interpreters. PMID- 17406960 TI - Association between ADHD and smoking in adolescence: shared genetic, environmental and psychopathological factors. AB - The present study aimed to examine the extent to which the co-occurrence of ADHD and smoking in adolescents could be attributed to common genetic, environmental and psychopathological factors. Data are from an ongoing prospective study of the outcome of early risk factors. At age 15 years, 305 adolescents completed self report questionnaires measuring tobacco consumption and deviant peer affiliations. Lifetime psychiatric diagnoses were obtained using standardized interviews. DNA was genotyped for the dopamine D4 receptor (DRD4) gene exon III polymorphism. Adolescents with a lifetime diagnosis of ADHD displayed significantly higher smoking activity than non-ADHD controls. A major component of this association could be accounted for by deviant peer affiliations and the comorbidity with oppositional-defiant and conduct disorder, while a minor part was attributable to DRD4 in males but not in females. These findings suggest that the association of ADHD with smoking relies on risk factors shared by the two behaviors. PMID- 17406961 TI - Modelling the roles of MAO and SSAO in glucose transport. AB - Amine oxidase substrates such as benzylamine and methylamine have been shown to stimulate glucose uptake by increasing the recruitment of the glucose transporter GLUT4 from vesicles within the cell to the cell surface. Inhibition of this effect by the presence of semicarbazide and catalase led to the suggestion that the process is mediated by the H(2)O(2) produced in the oxidation of these amines. Tyramine, which is a substrate for both MAO and SSAO, can also stimulate this process and in that case both MAO and SSAO inhibitors attenuate the effect. Benzylamine does not occur physiologically and tyramine is normally present in only very low amounts. We have suggested that adrenaline, which also stimulates glucose metabolism through adrenoceptors, may act as the physiological substrate for GLUT4 recruitment. It is a substrate for MAO but not SSAO. However, oxidation of adrenaline by MAO releases both H(2)O(2) and methylamine for further oxidation by SSAO. In order to gain a fuller understanding of this process we have performed simulation studies that may be used to assess the contributions of the amine oxidases to the process under a variety of conditions. The results are consistent with the experimentally observed behaviour. This approach not only helps to establish the feasibility of this process but also allows behaviour prediction and the identification of further experimental approaches. PMID- 17406962 TI - Ceruloplasmin and what it might do. AB - The blue coloured plasma protein ceruloplasmin binds up to 95% of circulating copper, and has several possible functions. It has been proposed to function in copper transport, oxidation of organic amines, iron(II) oxidation and the regulation of cellular iron levels, and catechols, radical scavenging and other antioxidant processes. This account will consider the relative importance of these multiple functions in terms of the physiological roles of ceruloplasmin. PMID- 17406963 TI - Hydrazine and amphetamine binding to amine oxidases: old drugs with new prospects. AB - Tranylcypromine (TCP), an amphetamine, is a reversible inhibitor of copper containing amine oxidases. We have solved the structure of the complex of TCP with the amine oxidase from E. coli (ECAO) and shown that only the (+)-enantiomer of TCP binds. Kinetic studies on 2-phenylethylamine and TCP binding to wild-type ECAO and mutational variants fully support the model in which binding of the protonated amine is the first step in the catalytic cycle. Hydrazines are irreversible inhibitors of copper-containing amine oxidases. Binding of hydrazines leads to an adduct ("Adduct 1") with a chromophore at 430 nm which converts at higher pH to another adduct ("Adduct 2") with a chromophore at 520 nm. We have determined the structures of Adduct 1 and 2 for 2-hydrazinopyridine reacted with ECAO. It has been found that Adduct 1 corresponds to the hydrazone and azo tautomers whilst Adduct 2 corresponds to the azo tautomer coordinated to the active site copper. The implications of these results in developing more specific drugs are discussed. PMID- 17406964 TI - Monoallelic expression of MAO-A in skin fibroblasts. AB - X chromosome inactivation in mammalian females occurs early in embryonic development and renders most genes on the inactive X chromosome transcriptionally silenced. As a consequence, females will display an X chromosomal parent-of origin mosaicism with regard to which parental allele that is expressed. Some genes however, escape inactivation and will therefore be expressed from both alleles. In this study we have investigated if the X-linked MAO-A gene have bi- or mono-allelic expression. This information would indicate whether or not MAO-A gene dosage could potentially explain the observed gender differences that show functional connections to the serotonin system, such as aggression and impulsiveness. To investigate the X inactivation status of MAO-A we have used primary clonal cell cultures, on which allelic expression was assessed with RFLP analysis. Our results show that the MAO-A gene has mono-allelic expression in these cells. This could have important implications for understanding traits that display gender differences. PMID- 17406965 TI - Semicarbazide-sensitive amine oxidase substrates fail to induce insulin-like effects in fat cells from AOC3 knockout mice. AB - Substrates of semicarbazide-sensitive amine oxidases (SSAO) stimulate glucose transport in adipocytes. To definitively demonstrate the involvement of SSAO in this insulin-like effect, glucose transport has been studied in fat cells from mice with a targeted deletion of AOC3, a gene encoding a SSAO called vascular adhesion protein-1. SSAO activity was present in white adipose tissues of wild type (WT) but was absent in AOC3KO mice. The SSAO-substrates benzylamine and methylamine were unable to stimulate hexose transport in adipocytes isolated from AOC3KO mice while they were active in WT adipocytes, especially in combination with vanadate. Impairment of amine-dependent glucose uptake was also observed with tyramine while there was no change in insulin responsiveness. These observations prove that the effects of exogenous or biogenic amines on glucose transport are not receptor-mediated but are oxidation-dependent. They also confirm that the major SSAO form expressed in mouse adipocytes is encoded by the AOC3 gene. PMID- 17406966 TI - Systematization of the vesical and uterovaginal efferences of the female inferior hypogastric plexus (pelvic): applications to pelvic surgery on women patients. AB - OBJECTIVE: To locate and describe the various efferences of the plexus in order to make it easier to avoid nerve lesions during pelvic surgery on women patients through a better anatomical knowledge of the inferior hypogastric plexus (IHP). MATERIALS AND METHODS: We dissected 27 formalin embalmed female anatomical subjects, none of which bore any stigmata of subumbilical surgery. The dissection was always performed using the same technique: identification of the inferior hypogastric plexus, whose posterior superior angle follows on from the hypogastric nerve and whose top, which is anterior and inferior, is located exactly at the ureter's point of entry into the base of the parametrium, underneath the posterior layer of the broad ligament. RESULTS: The IHP is located at the level of the posterior floor of the pelvis, opposite to the sacral concavity. Its top, which is anterior inferior, is at the point of contact with the ureter at its entry into the posterior layer of the broad ligament. The uterovaginal, vesical and rectal efferences originate in the paracervix. Three efferent nerves branch, two of them from its top and the third from its inferior edge: (1) A vaginal nerve, medial to the ureter, follows the uterine artery and divides into two groups: anterior thin, heading for the vagina and the uterus; posterior, voluminous, heading in a superior rectal direction (=superior rectal nerve). (2) A vesical nerve, lateral to the ureter, divides into two groups, lateral and medial. (3) The inferior rectal nerve emerges from the inferior edge of the IHP, between the fourth sacral root and the ureter's point of entry into the base of the parametrium. CONCLUSION: The ureter is the crucial point of reference for the IHP and its efferences and acts as a real guide for identifying the anterior inferior angle or top of the IHP, the origin of the vaginal nerve, the level of the ureterovesical junction and the division of the vesical nerve into its two medial and lateral branches. Dissecting underneath and inside the ureter and the uterine artery involves a risk of lesion of the vaginal nerve and its uterovaginal branches. Further forward, between the intersection and the ureterovesical junction, dissecting and/or coagulating under the ureter involves a risk of lesions to the vesical nerve, which are likely to explain the phenomena of denervation of the anterior floor encountered after certain hysterectomies and/or surgical treatments of vesicoureteral reflux. PMID- 17406967 TI - Anomalous hepatic arterial anatomy discovered during pancreaticoduodenectomy. AB - INTRODUCTION: A number of variations in hepatic arterial anatomy have been described. Anomalous arterial supply is of particular relevance to the hepatobiliary, pancreatic or liver transplant surgeon. CASE REPORT: We describe the case of a 57-year-old gentleman who presented with painless obstructive jaundice and was found to have a mass in the head of the pancreas. At pylorus preserving pancreaticoduodenectomy a distal quadrifurcation of the hepatic artery was discovered. It gave rise to the right hepatic artery, left hepatic artery, intermediate branch and a right accessory hepatic artery. The right accessory hepatic artery gave rise to the gastroduodenal artery. CONCLUSION: The surgeon and interventional radiologist need to be aware of the potential for such aberrant anatomy in order to avoid potentially disastrous complications. PMID- 17406968 TI - Stability of Nmi protein is controlled by its association with Tip60. AB - Tip60 exerts diverse biological functions through mechanisms that are either dependent or independent on its intrinsic histone acetyltransferase activity. In the present study, we identified Nmi (N-Myc and STATs Interactor) as a novel binding partner for Tip60 by a yeast two-hybrid screen. The association of Tip60 with Nmi was further confirmed by coimmunoprecipitation in mammalian cells. The zinc finger domain of Tip60 interacts with the NID repeats of Nmi, a region essential for the cytoplamic localization and homo- and heterodimerization of Nmi. We further showed that Nmi is an unstable protein and is targeted for proteasome-mediated degradation. The stability of Nmi can be enhanced by its association with Tip60, a process that is dependent on histone acetyltransferase activity of Tip60. The stabilization of Nmi by Tip60 is in part mediated by the translocation of Tip60 into cytoplasm to form distinct large cytoplasmic speckles. Our finding that Tip60 stabilizes Nmi through the formation of distinct cytoplasmic speckles provides a new mechanism to modulate Nmi-mediated functions. PMID- 17406969 TI - Effects of enhanced UV-B radiation on plant chemistry: nutritional consequences for a specialist and generalist lagomorph. AB - Ultraviolet-B (UV-B) radiation has been increasing in temperate latitudes in recent decades and is expected to continue rising for some time. Enhanced UV-B radiation can change plant chemistry, yet the effects of these changes on mammalian herbivores are unknown. To examine the influence of enhanced UV-B radiation on nutrition of a specialist and generalist hindgut fermenter, we measured nutritional and chemical constituents of three common North American range plants, big sagebrush (Artemisia tridentata), yarrow (Achillea millefolium), and bluebunch wheatgrass (Pseudoregneria spicata), and how these changes influenced in vitro dry matter digestibility and in vivo digestibility by pygmy rabbits (Brachylagus idahoensis) and eastern cottontails (Sylvilagus floridanus). Forages were irradiated for 3 mo with ambient (1x) or supplemental (1.6x) UV-B radiation representing a 15% ozone depletion for Pullman, WA, USA. Enhanced UV-B radiation had minimal effects on the nutritional content and the tannin-binding capacity of forages. Similarly, the terpene concentration in sagebrush and yarrow was not affected by higher UV-B irradiances. Flavonoid compounds increased in sagebrush but decreased in yarrow. Rabbit preference and intake was not affected by treatment levels for any forage species and no differences were found between treatments for dry matter, fiber, protein digestibility, and apparent digestible energy. PMID- 17406970 TI - Using system dynamics modeling to understand the impact of social change initiatives. AB - Community psychologists have a long history of interest in understanding social systems and how to bring about enduring positive change in these systems. However, the methods that community psychologists use to anticipate and evaluate the changes that result from system change efforts are less well developed. In the current paper, we introduce readers to system dynamics modeling, an action research approach to studying complex systems and the consequences of system change. We illustrate this approach by describing a system dynamics model of educational reform. We provide readers with an introduction to system dynamics modeling, as well as describe the strengths and limitations of the approach for application to community psychology. PMID- 17406971 TI - The system concept and systemic change: implications for community psychology. PMID- 17406972 TI - Personality and temperament correlates of pain catastrophizing in young adolescents. AB - Pain catastrophizing is generally viewed as an important cognitive factor underlying chronic pain. The present study examined personality and temperament correlates of pain catastrophizing in a sample of young adolescents (N = 132). Participants completed the Pain Catastrophizing Scale for Children, as well as scales for measuring sensitivity of the behavioral inhibition and behavioral activation systems (BIS-BAS), and various reactive and regulative temperament traits. Results demonstrated that BIS, reactive temperament traits (fear and anger-frustration), and perceptual sensitivity were positively related to pain catastrophizing, whereas regulative traits (attention control, inhibitory control) were negatively associated with this cognitive factor. Further, regression analyses demonstrated that only BIS and the temperamental traits of fear and perceptual sensitivity accounted for a unique proportion of the variance in adolescents' pain catastrophizing scores. PMID- 17406973 TI - Parent-assisted friendship training for children with autism spectrum disorders: effects of psychotropic medication. AB - Twenty-five 6 to 13-year-old children with autism spectrum disorders, who were high functioning, were given 12 weeks of parent-assisted children's friendship training. Thirteen were prescribed various psychotropic medications by physicians in the community prior to treatment (medicated) while 12 were not (unmedicated). Two parent-rated and three teacher-rated social measures served as outcome variables. Results revealed that unmedicated subjects had greater positive change on three of these five measures when compared to children in the medicated group. It was hypothesized that being prescribed psychotropic medication was a marker for refractory psychosocial treatment response by children with autism spectrum disorders. PMID- 17406974 TI - BRCA1 disease-associated haplotypes in Singapore Malay women with early-onset breast/ovarian cancer. PMID- 17406975 TI - Gender differences in the effects of prenatal stress on brain development and behaviour. AB - An increased incidence of anxiety, depression and attention deficits in children has been linked to psychological stress during pregnancy. Subjection of a pregnant rat to stress at a time when the foetal limbic and hypothalamic pituitary adrenal (HPA) axes develop results in anxiogenic and depressive behaviour and learning and attention deficits in the offspring, which depend on its gender, intensity and timing of the maternal stress and behaviour being tested. Maternal stress increases corticosterone levels in the foetal brain, decreases foetal testosterone and brain aromatase activity in males, and alters brain catecholamine activity to that in females. Learning deficits, reductions in hippocampal neurogenesis, LTP and dendritic spine density in the prefrontal cortex are more readily seen in prenatally-stressed males, while anxiety, depression and increased response of the HPA axis to stress are more prevalent in females. Genders may differ in the sensitivity of developing brain areas to stress hormones. PMID- 17406976 TI - Angiotensin receptor type 1 blockade in astroglia decreases hypoxia-induced cell damage and TNF alpha release. AB - The present study investigated the role of angiotensin receptors (AT-R) in the survival and inflammatory response of astroglia upon hypoxic injury. Exposure of rat astroglial primary cultures (APC) to hypoxic conditions (HC) led to decreased viability of the cells and to a 3.5-fold increase in TNF-alpha release. AT-R type1 (AT1-R) antagonist losartan and its metabolite EXP3174 decrease the LDH release (by 36 +/- 9%; 45 +/- 6%) from APC under HC. Losartan diminished TNF alpha release (by 40 +/- 15%) and the number of TUNEL-cells by 204 +/- 38% under HC, alone and together with angiotensin II (ATII), while EXP3174 was dependent on ATII for its effect on TNF-alpha. The AT2-R antagonist, PD123.319, did not influence the release of LDH and TNF-alpha under normoxic (NC) and HC. These data suggest that AT1-R may decrease the susceptibility of astrocytes to hypoxic injury and their propensity to release TNF-alpha. AT1-R antagonists may therefore be of therapeutic value during hypoxia-associated neurodegeneration. PMID- 17406977 TI - Beta-amyloid toxicity in embryonic rat astrocytes. AB - The senile plaques of Alzheimer's disease contain a high concentration of beta amyloid (betaA) protein, which may affect the glial population in the septal nucleus, an area of increased risk in AD. BetaA toxicity was measured in septal glia, via a dose-response experiment, by quantifying the effects of three different doses (0.1, 1, and 10 microM) of betaA on cell survival. Astrocytes from embryonic day-16 rats were grown in serum-free media in a single layer culture. Cells were treated on day in vitro (DIV)1 and survival was determined on DIV3 to ascertain which concentration was most toxic. In a separate set of experiments, an attempt was made to protect glial cells from the degenerative effects of betaA, with treatments of growth factors and estrogen. BetaA (10 microM) treatment was administered on DIV1, on DIV2 the cells were treated with estrogen (EST, 10 nM), insulin-like growth factors (IGF1 and IGF2, each 10 ng/ml), basic fibroblast growth factor (bFGF, 5 ng/ml) or nerve growth factor (NGF, 100 ng/ml), and on DIV3 the cells were visualized and quantified by fluorescence microscopy with DAPI (4,6-diamidino-2-phenylindole). In addition to dose-response and glial protection, experiments were also conducted to determine whether toxic effects were due to apoptosis. Our results suggest that the survival of glial populations is significantly affected in all three concentrations (0.1, 1.0, and 10 microM) of betaA. Glial protection was evident in the presence of NGF, for it showed the significantly highest survival rate relative to the betaA treatment alone. Furthermore, toxic effects of betaA appear to be due primarily to apoptosis. Significant reversal of betaA-induced apoptosis was seen with bFGF and IGF1. PMID- 17406978 TI - Neuroprotective effects of natural products: interaction with intracellular kinases, amyloid peptides and a possible role for transthyretin. AB - Various studies reported on the neuroprotective effects of natural products, particularly polyphenols, widely present in food and beverages. For example, we have shown that resveratrol, a polyphenol contained present in red wine and other foods, activates the phosphorylation of protein kinase C (PKC), this effect being involved in its neuroprotective action against Ass-induced toxicity. Moreover, tea-derived catechin gallate esters inhibit the formation Ass oligomers/fibrils, suggesting that this action likely contributes to their neuroprotective effects. Interestingly, the effects of polyphenols may be attributable, at least in part, to the presence of specific binding sites. Autoradiographic studies revealed that these binding sites are particularly enriched in choroids plexus in the rat brain. Interestingly, the choroid plexus secretes transthyretin, a protein that has been shown to prevent Abeta aggregation and that may be critical to the maintenance of normal learning capacities in aging. Taken together, these data suggest that polyphenols target multiple enzymes/proteins leading to their neuroprotective actions. PMID- 17406979 TI - Life-long hippocampal neurogenesis: environmental, pharmacological and neurochemical modulations. AB - It is now well documented that active neurogenesis does exist throughout the life span in the brain of various species including human. Two discrete brain regions contain progenitor cells that are capable of differentiating into neurons or glia, the subventricular zone and the dentate gyrus of the hippocampal formation. Recent studies have shown that neurogenesis can be modulated by a variety of factors, including stress and neurohormones, growth factors, neurotransmitters, drugs of abuse, and also strokes and traumatic brain injuries. In particular, the hippocampal neurogenesis may play a role in neuroadaptation associated with pathologies, such as cognitive disorders and depression. The increased neurogenesis at sites of injury may represent an attempt by the central nervous system to regenerate after damage. We herein review the most significant data on hippocampal neurogenesis in brain under various pathological conditions, with a special attention to mood disorders including depression and addiction. PMID- 17406980 TI - Fas antigen and sporadic Alzheimer's disease in Southern Italy: evaluation of two polymorphisms in the TNFRSF6 gene. AB - Tumor Necrosis Factor Receptor Super Family 6 gene (TNFRSF6), also known as FAS, encodes the Fas antigen, a cell surface receptor mediating cell apoptosis, situated on chromosome 10q located near the region of linkage to sporadic Alzheimer's disease (sAD). FAS levels have been reported elevated in the brain of AD patients. Due to both positional and pathobiological criteria, the association of the FAS antigen with this pathology is of great interest. We have tested two SNPs in the FAS gene in 223 Italian patients with non-familial AD from Southern Italy (Calabria region) and 211 healthy control subjects. No significant differences in allelic and genotypic distributions were found between cases and controls, or late and early-onset AD patients, thus suggesting that these polymorphisms do not represent an AD risk factor in our population. PMID- 17406981 TI - Regional distribution of human trypsinogen 4 in human brain at mRNA and protein level. AB - Gene PRSS3 on chromosome 9 of the human genome encodes, due to alternative splicing, both mesotrypsinogen and trypsinogen 4. Mesotrypsinogen has long been known as a minor component of trypsinogens expressed in human pancreas, while the mRNA for trypsinogen 4 has recently been identified in brain and other human tissues. We measured the amount of trypsinogen 4 mRNA and the quantity of the protein as well in 17 selected areas of the human brain. Our data suggest that human trypsinogen 4 is widely but unevenly distributed in the human brain. By immunohistochemistry, here we show that this protease is localized in neurons and glial cells, predominantly in astrocytes. In addition to cellular immunoreactivity, human trypsinogen 4 immunopositive dots were detected in the extracellular matrix, supporting the view that human trypsinogen 4 might be released from the cells under special conditions. PMID- 17406982 TI - Effect of electroacupuncture on neurotrophin expression in cat spinal cord after partial dorsal rhizotomy. AB - Neuroplasticity of the spinal cord following electroacupuncture (EA) has been demonstrated although little is known about the possible underlying mechanism. This study evaluated the effect of EA on expression of neurotrophins in the lamina II of the spinal cord, in cats subjected to dorsal rhizotomy. Cats received bilateral removal of L1-L5 and L7-S2 dorsal root ganglia (DRG, L6 DRG spared) and unilateral EA. They were sacrificed 7 days after surgery, and the L6 spinal segment removed and processed by immunohistochemistry and in situ hybridization histochemistry, to demonstrate the expression of neurotrophins. Significantly greater numbers of nerve growth factor (NGF) and neurotrophin-3 (NT 3) positive neurons, brain-derived neurotrophic factor (BDNF) immunoreactive varicosities and NT-3 positive neurons and glial cells were observed in lamina II on the acupunctured (left) side, compared to the non-acupunctured, contralateral side. Greater number of neurons expressing NGF mRNA was also observed on the acupunctured side. No signal for mRNA to BDNF and NT-3 was detected. The above findings demonstrate that EA can increase the expression of endogenous NGF at both the mRNA and protein level, and BDNF and NT-3 at the protein level. It is postulated that EA may promote the plasticity of the spinal cord by inducing increased expression of neurotrophins. PMID- 17406983 TI - Class III beta-tubulin and gamma-tubulin are co-expressed and form complexes in human glioblastoma cells. AB - We have previously shown that the neuronal-associated class III beta-tubulin isotype and the centrosome-associated gamma-tubulin are aberrantly expressed in astrocytic gliomas (Cell Motil Cytoskeleton 2003, 55:77-96; J Neuropathol Exp Neurol 2006, 65:455-467). Here we determined the expression, distribution and interaction of betaIII-tubulin and gamma-tubulin in diffuse-type astrocytic gliomas (grades II-IV) (n = 17) and the human glioblastoma cell line T98G. By immunohistochemistry and immunofluorescence microscopy, betaIII-tubulin and gamma tubulin were co-distributed in anaplastic astrocytomas and glioblastomas and to a lesser extent, in low-grade diffuse astrocytomas (P < 0.05). In T98G glioblastoma cells betaIII-tubulin was associated with microtubules whereas gamma-tubulin exhibited striking diffuse cytoplasmic staining in addition to its expectant centrosome-associated pericentriolar distribution. Treatment with different anti microtubule drugs revealed that betaIII-tubulin was not associated with insoluble gamma-tubulin aggregates. On the other hand, immunoprecipitation experiments unveiled that both tubulins formed complexes in soluble cytoplasmic pools, where substantial amounts of these proteins were located. We suggest that aberrant expression and interactions of betaIII-tubulin and gamma-tubulin may be linked to malignant changes in glial cells. PMID- 17406984 TI - Basic fibroblast growth factor prevents the memory impairment induced by gastrin releasing peptide receptor antagonism in area CA1 of the rat hippocampus. AB - Increasing evidence indicates that the gastrin-releasing peptide receptor (GRPR) is implicated in regulating synaptic plasticity and memory formation in the hippocampus and other brain areas. However, the molecular mechanisms underlying the memory-impairing effects of GRPR antagonism have remained unclear. Here we report that basic fibroblast growth factor (bFGF/FGF-2) rescues the memory impairment induced by GRPR antagonism in the rat dorsal hippocampus. The GRPR antagonist [D-Tpi(6), Leu(13) psi(CH(2)NH)-Leu(14)] bombesin (6-14) (RC-3095) at 1.0 microg impaired, whereas bFGF at 0.25 microg enhanced, 24 h retention of inhibitory avoidance (IA) when infused immediately after training into the CA1 hippocampal area in male rats. Coinfusion with an otherwise ineffective dose of bFGF blocked the memory-impairing effect of RC-3095. These findings suggest that the memory-impairing effects of GRPR antagonists might be partially mediated by an inhibition in the function and/or expression of neuronal bFGF or diminished activation of intracellular protein kinase pathways associated with bFGF signaling. PMID- 17406985 TI - Heavy metals modulate glutamatergic system in human platelets. AB - Research strategies have been developed to characterize parameters in peripheral tissues that might easily be measured in humans as surrogate markers of damage, dysfunction or interactions involving neural targets of toxicants. The similarities between platelet and neuron may even be clinically important, as a number of biochemical markers show parallel changes in the central nervous system (CNS) and platelets. The purpose of our research was to investigate the effect of Hg(2+), Pb(2+) and Cd(2+) on the [(3)H]-glutamate binding and [(3)H]-glutamate uptake in human platelets. The involvement of oxidative stress in the modulation of glutamatergic system induced by heavy metals was also investigated. The present study clearly demonstrates that Hg(2+), Cd(2+), and Pb(2+) inhibited [(3)H]-glutamate uptake in human platelets. Hg(2+) inhibited [(3)H]-glutamate binding, while Cd(2+) and Pb(2+) stimulated [(3)H]-glutamate binding in human platelets. Hg(2+), Cd(2+) and Pb(2+) increased lipid peroxidation levels and reactive oxygen species (ROS) measurement in platelets. The present limited results could suggest that glutamatergic system may be used as a potential biomarker for neurotoxic action of heavy metals in humans. PMID- 17406986 TI - Establishment of a novel orthotopic xenograft model of human gallbladder carcinoma. AB - OBJECTIVE: Gallbladder cancer is characterized by high morbidity and mortality. An appropriate human xenograft animal model could serve as a research tool to investigate new therapeutic strategies. SUMMARY BACKGROUND DATA: To date, the few reports describing a xenograft animal model showed significant limitations. We improved a murine orthotopic human xenotransplantation model by implanting human gallbladder carcinoma cells directly into the lumen of the gallbladder. METHODS: Mz-ChA-1 cells were orthotopically injected into the gallbladder of Severe Combined Immune Deficiency (SCID) beige mice inducing the growth of solid tumors. The natural course of the disease, tumor growth, and metastases were analyzed. The cytotoxic drug gemcitabine was tested in vitro and in vitro. RESULTS: All animals revealed solid tumors in the inoculated area with liver infiltration. The median tumor volume in the untreated group was significantly higher than in the gemcitabine-treated group. Immunohistochemical staining revealed expression of human cytokeratin 7 and cytokeratin 8. To analyze tumor cell proliferation, the tumors were stained for the antigen Ki-67, and labeling indices were calculated for both groups. Animals receiving gemcitabine treatment showed significantly lower mean labeling indices. In vitro investigation revealed a significant reduction of DNA synthesis. DNA fragmentation, as a measure of apoptosis, was elevated by roughly 20% within 24 h of treatment. With this, we successfully established an orthotopic xenotransplant animal model and investigated the in vitro and in vivo effects of gemcitabine in human xenografted Mz-ChA-1 gallbladder adenocarcinoma. CONCLUSION: This model resembles the clinical situation as closely as possible and offers a relevant option for the preclinical testing of new therapeutic strategies. PMID- 17406987 TI - Summative software evaluation of a therapeutic guideline assistance system for empiric antimicrobial therapy in ICU. AB - OBJECTIVE: While developing the patient data management system ICUData in close cooperation with the software company (IMESO GmbH, Huttenberg, Germany), a therapeutic guideline assistance system for empiric antimicrobial therapy in ICU (called "Antibiotic Wizard") could be introduced and integrated into the existing software. After its introduction into clinical routine, the first version was to be tested, checked for usability and compared to other software products with the help of the IsoMetrics(s) inventory (based on the EN ISO 9241-10 for computer assisted workflows). METHODS: Half a year after introducing the "Antibiotic Wizard" in the ICUs, 40 physicians from different specialties at different levels of training were surveyed in order to detect deficiencies in the use of the program. The results of these surveys were compared to surveys on the word processing software Word for Windows (WinWord) from Microsoft, the hospital information system IS-H*MED from SAP (online and paper surveys) and the administrative program, SAP R/3 HR, also from SAP. RESULTS: Reliabilities (Cronbach's Alpha) of the subscales ranged from satisfactory (alpha > 0.70) to good (alpha > 0.80), except for "Controllability" (alpha = 0.663) and "Error tolerance" (alpha = 0.693). Medians for individual subscales ranged between 3.04 ("Error tolerance") and 3.96 ("Suitability for learning"). The "Antibiotic Wizard" showed significantly better results compared to both IS-H*MED and SAP R/3 HR in the subscales of "Suitability for the task", "Self-descriptiveness" and "Suitability for learning". In contrast, "Self-descriptiveness" "Controllability" and "Error tolerance" were significantly worse compared to WinWord. CONCLUSIONS: In generally, the usability of the "Antibiotic Wizard" was deemed good. Some weaknesses were found in the fields of "Error tolerance" and "Controllability". These problems will be corrected in future versions. PMID- 17406988 TI - Use of radio frequency identification (RFID) tags in bedside monitoring of endotracheal tube position. AB - Improper positioning of the endotracheal tube during intubation poses a serious health risk to patients. In one prospective study of 219 critically ill patients, 14% required endotracheal tube repositioning after intubation [Brunel et al. Chest 1989; 96: 1043-1045] While a variety of techniques are used to confirm proper tube placement, a chest X-ray is usually employed for definitive verification. Radio frequency identification (RFID) technology, in which an RFID reader emits and receives a signal from an RFID tag, may be useful in evaluating endotracheal tube position. RFID technology has already been approved for use in humans as a safe and effective tool in a variety of applications. The use of handheld RFID detectors and RFID tag-labeled endotracheal tubes could allow for easy and accurate bedside monitoring of endotracheal tube position, once initial proper placement is confirmed. PMID- 17406989 TI - Incidence of esophageal and gastric cancers among Hispanics, non-Hispanic whites and non-Hispanic blacks in the United States: subsite and histology differences. AB - OBJECTIVE: We examined subsite- and histology-specific esophageal and gastric cancer incidence patterns among Hispanics/Latinos and compared them with non Hispanic whites and non-Hispanic blacks. METHODS: Data on newly diagnosed esophageal and gastric cancers for 1998-2002 were obtained from 37 population based central cancer registries, representing 66% of the Hispanic population in the United States. Age-adjusted incidence rates (2000 US) were computed by race/ethnicity, sex, anatomic subsite, and histology. The differences in incidence rates between Hispanics and non-Hispanics were examined using the two tailed z-statistic. RESULTS: Squamous cell carcinoma accounted for 50% and 57% of esophageal cancers among Hispanic men and women, respectively, while adenocarcinoma accounted for 43% among Hispanic men and 35% among Hispanic women. The incidence rate of squamous cell carcinoma was 48% higher among Hispanic men (2.94 per 100,000) than non-Hispanic white men (1.99 per 100,000) but about 70% lower among Hispanics than non-Hispanic blacks, for both men and women. In contrast, the incidence rates of esophageal adenocarcinoma were lower among Hispanics than non-Hispanic whites (58% lower for men and 33% for women) but higher than non-Hispanic blacks (70% higher for men and 64% for women). Cardia adenocarcinoma accounted for 10-15% of gastric cancers among Hispanics, and the incidence rate among Hispanic men (2.42 per 100,000) was 33% lower than the rate of non-Hispanic white men (3.62 per 100,000) but 37% higher than that of non Hispanic black men. The rate among Hispanic women (0.86 per 100,000), however, was 20% higher than that of non-Hispanic white women (0.72 per 100,000) and 51% higher than for non-Hispanic black women. Gastric non-cardia cancer accounted for approximately 50% of gastric cancers among Hispanics (8.32 per 100,000 for men and 4.90 per 100,000 for women), and the rates were almost two times higher than for non-Hispanic whites (2.95 per 100,000 for men and 1.72 per 100,000 for women) but about the same as the non-Hispanic blacks. CONCLUSION: Subsite- and histology specific incidence rates of esophageal and gastric cancers among Hispanics/Latinos differ from non-Hispanics. The incidence rates of gastric non cardia cancer are almost two times higher among Hispanics than non-Hispanic whites, both men and women. The rates of gastric cardia cancer are lower among Hispanics than non-Hispanic whites for men but higher for women. The rates of esophageal and gastric cardia adenocarcinomas are higher among Hispanics than non Hispanic blacks. PMID- 17406990 TI - Nucleolus and chromosome relationships at pachynema in four Scarabaeoidea (Coleoptera) species with various combinations of NOR and sex chromosomes. AB - Nucleolus organizer regions (NORs) and nucleolus locations were studied after silver staining in spermatocytes at pachynema from four beetle species selected for their various combinations of sex chromosomes. Their karyotypic formulae were: 18,neoXY (Dorcus parallelipipedus); 25,X (Passalus unicornis) and 20,Xyp (Cetonia aurata and Protaecia (Potosia) opaca). NORs were located in the short arms of a unique acrocentric autosome pair in the first three and in intercalary position in a sub-metacentric autosome pair in the last species. Silver staining gave remarkably more consistent results in pachytene than in mitotic spreads, enabling the detection of both NORs and nucleoli, and also better results in embryo than in spermatogonial metaphases. At pachynema the NORs were elongated, roughly in proportion to the number of nucleoli, which always remained associated with NOR. Nucleoli were not recurrently associated with sex chromosomes, except in P. unicornis, at late pachynema. In C. aurata and P. opaca the sex body was recurrently associated with acrocentric short arms and metacentric telomeres, respectively. Even in these simple situations, with NORs located in a single autosome pair, the number of nucleoli and their relationships with sex chromosomes varied strongly from species to species. These variations appear to be largely determined by the chromosome rearrangements which have occurred during evolution, which makes extrapolations and generalizations quite hazardous. In D. parallelipipedus pachytene cells a quasi-systematic and transient fusion between the terminal heterochromatin of two sub-metacentrics was detected. Other chromosome bivalents could also be occasionally associated, but not the NOR carrier one. A strong enhancement of DAPI or quinacrine mustard staining was observed at the fusion point. PMID- 17406991 TI - Integrated cytogenetic BAC map of the genome of the gray, short-tailed opossum, Monodelphis domestica. AB - The generation of high-quality genome assemblies for numerous species is advancing at a rapid pace. As the number of genome assemblies increases, so does our ability to investigate genome relationships and their contributions to unraveling complex biological, evolutionary, and biomedical processes. A key process in the generation of a genome assembly is to determine and verify the precise physical location and order of the large sequence blocks (scaffolds) that result from the assembly. For organisms of relatively recent common ancestry this process may be achieved largely through comparative sequence alignment. However, as the evolutionary distance between species lengthens, the use of comparative sequence alignment becomes increasingly less reliable. Simultaneous cytogenetic mapping, using multicolor fluorescence in-situ hybridization (FISH) analysis, offers an alternative means to define the cytogenetic location and relative order of DNA sequences, thereby anchoring the genome sequence to the karyotype. In this article we report the molecular cytogenetic locations of 415 bacterial artificial chromosome (BAC) clones that served to anchor sequence scaffolds of the gray, short-tailed opossum (Monodelphis domestica) to its karyotype, which enabled accurate integration of these regions into the genome assembly. PMID- 17406992 TI - A new platform linking chromosomal and sequence information. AB - We have tested whether a direct correlation of sequence information and staining properties of chromosomes is possible and whether this combined information can be used to precisely map any position on the chromosome. Despite huge differences of compaction between the naked DNA and the DNA packed in chromosomes we found a striking correlation when visualizing the GGCC density on both levels. Software was developed that allows one to superimpose chromosomal fluorescence intensity profiles generated by chromolysin A3 (CMA3) staining with GGCC density extracted from the Ensembl database. Thus, any position along the chromosome can be defined in megabase pairs (Mb) besides the cytoband information, enabling direct alignment of chromosomal information with the sequence data. The mapping tool was validated using 13 different BAC clones, resulting in a mean difference from Ensembl data of 2 Mb (ranging from 0.79 to 3.57 Mb). Our results indicate that the sequence density information and information gained with sequence-specific fluorochromes are superimposable. Thus, the visualized GGCC motif density along the chromosome (sequence bands) provides a unique platform for comparing different types of genomic information. PMID- 17406993 TI - DNA content of the B chromosomes in grasshopper Podisma kanoi Storozh. (Orthoptera, Acrididae). AB - A DNA library derived from the B chromosome of Podisma kanoi was obtained by chromosome microdissection. A total of 153 DNA clones were isolated from the microdissected DNA library. Twenty of them were sequenced. A comparison of B chromosome DNA sequences with sequences of other species from the DDBJ/GenBank/EMBL database ( http://www.ddbj.nig.ac.jp/ ) was performed. Different patterns of signals were observed after FISH with labeled cloned DNA fragments. FISH signals with cloned DNA fragments painted either whole Bs or their different regions. Some clones also gave signals in pericentromeric regions of A chromosomes. Other cloned DNA fragments gave only background-like signals on A and B chromosomes. Comparative FISH analysis of B chromosomes in Podisma kanoi and P. sapporensis with DNA probes derived from the Bs of these species revealed homologous DNA that was confined within pericentromeric and telemetric regions of the B chromosome in P. kanoi. In contrast to the B chromosomes in P. sapporensis containing large regions enriched with rDNA, only a small cluster of rDNA was detected in one of the examined B chromosomes in P. kanoi. The data strongly suggest an independent origin of B chromosomes in two closely related Podisma species. PMID- 17406994 TI - The polycomb group protein SUZ12 regulates histone H3 lysine 9 methylation and HP1 alpha distribution. AB - Regulation of histone methylation is critical for proper gene expression and chromosome function. Suppressor of Zeste 12 (SUZ12) is a requisite member of the EED/EZH2 histone methyltransferase complexes, and is required for full activity of these complexes in vitro. In mammals and flies, SUZ12/Su(z)12 is necessary for trimethylation of histone H3 on lysine 27 (H3K27me3) on facultative heterochromatin. However, Su(z)12 is unique among Polycomb Group Proteins in that Su(z)12 mutant flies exhibit gross defects in position effect variegation, suggesting a role for Su(z)12 in constitutive heterochromatin formation. We investigated the role of Suz12 in constitutive heterochromatin and discovered that Suz12 is required for histone H3 lysine 9 tri-methylation (H3K9me3) in differentiated but not undifferentiated mouse embryonic stem cells. Knockdown of SUZ12 in human cells caused a reduction in H3K27me3 and H3K9me3, and altered the distribution of HP1 alpha. In contrast, EZH2 knockdown caused loss of H3K27me3 but not H3K9me3, indicating that SUZ12 regulates H3-K9 methylation in an EZH2 independent fashion. This work uncovers a role for SUZ12 in H3-K9 methylation. PMID- 17406995 TI - Monitoring and assessment of toxic metals in Gulf War oil spill contaminated soil using laser-induced breakdown spectroscopy. AB - Laser-induced breakdown spectroscopy (LIBS) was applied for the detection of toxic metals in oil spill contaminated soil (OSCS). The OSCS samples were collected from Khursania Saudi Arabia along the coast of Persian Gulf exposed to oil spills in 1991 Gulf war. Environmentally important elements like Aluminum Magnesium, Calcium, Chromium, Titanium, Strontium, Iron, Barium, Sodium, potassium, Zirconium and Vanadium from the contaminated soil have been detected. Optimal experimental conditions for analysis were investigated. The LIBS system was calibrated using standard samples containing these trace elements. The results obtained using Laser-Induced Breakdown Spectroscopy (LIBS) were compared with the results obtained using Inductively Coupled Plasma Emission Spectroscopy (ICP). The concentrations of some elements (Ba and Cr) were found higher than permissible safe limits. Health risks associated with exposure to such toxic elements are also discussed. PMID- 17406996 TI - Status of insecticide contamination of soil and water in Haryana, India. AB - Twelve samples each of soil and ground water were collected from paddy-wheat, paddy-cotton, sugarcane fields and tube wells from same or near by fields around Hisar, Haryana, India during 2002-2003 to monitor pesticide residues. Residues were estimated by GC-ECD and GC-NPD systems equipped with capillary columns for organochlorine, synthetic pyrethroid and organophosphate insecticides. In soil, HCH (0.002-0.051 microg g(-1)), DDT (0.001-0.066 microg g(-1)), endosulfan (0.002 0.039 microg g(-1)) and chlordane (0.0002-0.019 microg g(-1)) among organochlorines, cypermethrin (0.001-0.035 microg g(-1)) and fenvalerate (0.001 0.022 microg g(-1)) among synthetic pyrethroids and chlorpyriphos (0.002-0.172 microg g(-1)), malathion (0.002-0.008 microg g(-1)), quinalphos (0.001-0.010 microg g(-1)) among organophosphates were detected. Dominant contaminants were DDT, cypermethrin and chlorpyriphos from the respective groups. In water samples, HCH, DDT, endosulfan and cypermethrin residues were observed frequently. Only chlorpyriphos among organophosphates was detected in 10 samples. On consideration of tube well water for drinking purpose, about 80% samples were found to contain residues above the regulatory limits. PMID- 17406997 TI - Accumulations of total metal in dominant shrimp species (Palaemon adspersus, Palaemon serratus, Parapenaeus longirostris) and bottom surface sediments obtained from the Northern Inner Shelf of the Sea of Marmara. AB - The aim of this study is to determine the total metal (aluminum, copper, manganese, nickel, lead, zinc, cadmium, iron, mercury) contents of dominant shrimp species and sediments present at 1-50 m depths of the Northern Inner Shelf of the Sea of Marmara. Shrimp and sediment samples were collected from four regions (Buyukcekmece, Silivri, Tekirdag, Sarkoy) and from different depths (1, 5, 10, 20, 30, 50 m) at each region in September and November of 2003. Three shrimp species [Palaemon adspersus (Rathke, 1937), Palaemon serratus (Pennant, 1777), Parapenaeus longirostris (H. Lucas, 1846)] were identified to be dominant as a result of the examination on the obtained samples. Heavy metal contents of these three species were determined and the results were compared with the acceptable treshold values of the Seafood Standards and also with available literatures. The Cu contents were found to be higher than the treshold limits in all samples except P. longirostris from Silivri, also the Cd contents in all samples except P. adspersus from Buyukcekmece, the Zn contents only in P. adspersus and P. longirostris from Tekirdag, and the Pb contents in all species from all regions. These high values are the indicator of industrial pollution. There is not any data in the Seafood Standards about Mn, Ni, Fe and Al contents in shrimp species. The values of these metal contents were given in the present study. The examination of total metal distributions in bottom surface sediment samples in the Northern Inner Shelf of the Sea of Marmara showed that the determined values were higher than the shale average at some depths of examined stations. PMID- 17406998 TI - Structure-toxicity relationships of thiono and seleno phosphoramidate compounds: new type of acetylcholinesterase inhibitors. AB - Novel thiono and seleno phosphoramidate compounds with the general formula (X)(Y)P(C(6)H(5))(2); (X = NMe(2) & Y = S, 1a; X = NEt(2) & Y = S, 2a; X = NMe(CH(2)Ph) & Y = S, 3a; X = NH(CH(2)Ph) & Y = S, 4a; X = NEt(CH(2)Ph) & Y = S, 5a; X = N(C(Me)(3)) (CH(2)Ph) & Y = S, 6a; X = N(CH(2)Ph)(2) & Y = S, 7a; X = NMe(2) & Y = Se, 1b; X = NEt(2) & Y = Se, 2b; X = NMe(CH(2)Ph) & Y = Se, 3b; X = NH (CH(2)Ph) & Y = Se, 4b; X = NEt(CH(2)Ph) & Y = Se, 5b; X = N(C(Me)(3))(CH(2)Ph) & Y = Se, 6b and X = N(CH(2)Ph)(2) & Y = Se, 7b) were prepared and characterized by (1)H, (31)P and( 13)C NMR and IR spectroscopy and elemental analysis. (31)P chemical shift of thiono and seleno derivatives didn't show significant different because of their little difference in electronegativity sulfur and selenium. Hydrophobic parameter of compounds was determined by measurement of octanol-water partition coefficient by shake-flask technique. Determination of human erythrocyte acetylcholinesterase (hAChE) activity was carried out according to the Ellman's modified kinetic method. IC(50 )values of the selected thiono and seleno compounds varied from 3.4 to 0.11 and 9.9 to 5.1 mM, respectively. The seleno compounds show lower affinities for hAChE relative to the thino compounds. These results demonstrate that hydrophobic and electronic factors of the organophosphorus compounds play a key role on the inhibitory potency. PMID- 17406999 TI - A simple approach to the synthesis of highly functionalized 3-alkylidene-2,3 dihydro-1H-pyrrole-2-ol derivatives and related pyrroles. AB - The reaction of various primary amines with acetylenic esters at ambient temperature in THF/H(2)O (50:50) produced related enaminones. Subsequently reaction of these in situ prepared enaminones with dibenzoylacetylene leads to 3 alkylidene-2,3-dihydro-1H-pyrrol-2-ol derivatives in good yields. The reaction of these products with various alcohols in the presence of a catalytic amount of HCl produced highly functionalized pyrroles in nearly quantitative yields. PMID- 17407000 TI - The Streptomyces violaceusniger clade: a home for Streptomycetes with rugose ornamented spores. AB - The taxonomic status of 16 strains received as Streptomyces hygroscopicus, Streptomyces melanosporofaciens, Streptomyces sparsogenes, Streptomyces sporoclivatus and Streptomyces violaceusniger was evaluated in a polyphasic study. Eleven of the organisms formed a distinct clade in the Streptomyces 16S rRNA gene tree with the type strains of Streptomyces asiaticus, Streptomyces cangkringensis, Streptomyces indonesiensis, Streptomyces javensis, Streptomyces malaysiensis, Streptomyces rhizosphaericus, Streptomyces yatensis and Streptomyces yogyakartensis, the members of this group produced rugose ornamented spores in spiral spore chains. The eleven strains were assigned to three established and four novel species, namely Streptomyces albiflaviniger sp. nov., Streptomyces demainii sp. nov., Streptomyces geldanamycininus sp. nov., Streptomyces griseiniger sp. nov., and Streptomyces hygroscopicus, Streptomyces melanosporofaciens and Streptomyces violaceusniger. It is also proposed that S. sporoclivatus becomes a subjective synonym of S. melanosporofaciens. S. sparsogenes NRRL 2940(T), which produced ridged ornamented spores in spiral spore chains, formed a distinct phyletic line in the Streptomyces 16S rRNA gene tree and was readily distinguished from the other strains using a range of phenotypic properties. S. violaceusniger strains NRRL 8097, NRRL B-5799, NRRL 2834 and ISP 5182 fell outside the S. violaceusniger 16S rRNA gene clade and formed either smooth or ridged ornamented spores in either flexuous or spiral spore chains. These organisms were distinguished from one another and from their closest phylogenetic neighbors and were considered to merit species status as Streptomyces auratus sp. nov., Streptomyces phaeoluteichromatogenes sp. nov., Streptomyces phaeogriseichromatogenes sp. nov., and Streptomyces phaeoluteigriseus sp. nov., respectively. PMID- 17407001 TI - Low citrulline may not be diagnostic of ornithine transcarbamylase deficiency: a case report. AB - A newborn boy with family history of severe ornithine transcarbamylase (OTC) deficiency was investigated prospectively and managed aggressively at birth based on an existing protocol for at risk neonates. Undetectable citrulline levels at birth suggested that the infant was affected; however, normal plasma glutamine and urine orotic acid levels confused the diagnosis to some extent. Mutation testing confirmed that the patient did not have OTC deficiency. Thus the low plasma citrulline level did not validate our initial biochemical suspicion of OTC deficiency, and this highlights the importance of considering all available clinical, biochemical and molecular evidence in determining disease status. PMID- 17407002 TI - Serum lipid and lipoprotein profile of patients with glycogen storage disease types I, III and IX. AB - With current dietary therapy, life expectancy in glycogen storage disease (GSD) has improved considerably and more children reach adulthood. Notwithstanding intensive dietary therapy, moderate to severe hyperlipidaemia is still observed frequently. There is limited information about the type and extent of hyperlipidaemia. We studied the lipid profile in 20 patients, aged 8-54 years, of the three (types I, III and IX) most common forms of adult GSD. Hyperlipidaemia was shown to be type-specific, affecting predominantly patients with GSD type Ia, who showed marked combined hypercholesterolaemia and hypertriglyceridaemia. By contrast, a heterogeneous distribution of HDL was found in patients with GSD I and III. There was no significant difference in Apo Al and Apo B concentrations between groups. In addition, mass measurements of the fractions of VLDL1, VLDL2 and IDL were raised in all patients with GSD Ia by comparison with all other patients with GSD. Patients with GSD type Ia have lipid concentrations and individual mass measurements that are consistent with ranges found in patients who have a significant risk of atherosclerosis. Accumulated evidence, however, suggest GSD type Ia patients do not have an increased risk of atherosclerotic cardiovascular disease (CVD) but the reason remains unknown. Intervention to reduce their lipid levels could therefore be on the basis of seeking to prevent the risk of pancreatitis rather than that of CVD. PMID- 17407003 TI - Pathogenicity of fungal isolates (Ascomycota: Hypocreales) against Peregrinus maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae), vectors of corn diseases. AB - Preliminary screening assays were carried out on 17 isolates from five fungal species Beauveria bassiana, Lecanicillium muscarium, Metarhizium anisopliae, Isaria farinosa, and I. fumosorosea. The three most effective isolates against Peregrinus maidis (Hemiptera: Delphacidae) were B. bassiana CEP 147, CEP 150, and CEP 189. There were no consistent differences found in males and females regarding fungal susceptibility. However, more females than males were proportionally infected. There was not a correlation between the percentage of conidial germination and the percentage of mortality caused by fungal infection in any of the treatments. Only B. bassiana CEP 147, which caused a cumulative mortality of 69.8 +/- 6.4% after 7 days post-inoculation, was selected to be assayed against adults of P. maidis, Delphacodes kuscheli (Hemiptera: Delphacidae), and Dalbulus maidis (Hemiptera: Cicadellidae). In pathogenicity tests significant differences were observed among treatments. After 2 weeks post inoculation, both D. kuscheli (cumulative mortality of 73.3 +/- 9.0%) and P. maidis (cumulative mortality of 68.6 +/- 6.7%) were significantly more susceptible than D. maidis (cumulative mortality of 49.9 +/- 9.7%) to the selected isolate. PMID- 17407004 TI - Phylogeny and biogeography of Rhabdochona Railliet, 1916 (Nematoda: Rhabdochonidae) species from the Americas. AB - A phylogenetic analysis of 40 species of Rhabdochona Railliet, 1916, including all 21 valid species in the Americas, resulted in 1733 equally most parsimonious trees and indicates that Rhabdochona is arguably monophyletic. Species from the Americas do not form a monophyletic group, since each of the six clades of Rhabdochona includes species from the Americas and species from other continents. The synapomorphies defining each clade stem from the morphology of the left spicule. Teeth number was consistent in one clade only, suggesting that this character, while useful for taxonomic purposes, is not indicative of phylogeny. Species of Rhabdochona associated with certain host groups, such as salmonids, catostomids and goodeids, do not always form monophyletic assemblages, nor do species associated with smaller discrete areas, such as the Mesa Central of Mexico. This indicates widespread host-switching rather than co-speciation as the main phenomenon in the evolution of this group, at least in the species from the Americas. Phylogenetic patterns reveal an ancient origin for the group that probably pre-dates current continental configurations. PMID- 17407007 TI - [Localization of lacrimal drainage system obstruction by diaphanoscopy]. AB - Endonasal dacryocystorhinostomy is the standard treatment for obstruction of the lacrimal drainage system. Localization of the stenosis can be difficult intraoperatively, especially in chronic sclerosing dacryocystitis or re operations. We present a technique of intraoperative diaphanoscopic localization of the stenosis with the use of a special small light fiber that is inserting through the lacrimal points down to the stenosis. The lighted fiber illuminates the area, which has to be exposed endonasally and drain to the nose. This little addition contributes to a meticulous and straightforward operative technique. PMID- 17407008 TI - [Automation in surgery: a systematical approach]. AB - Surgical assistance systems permit a misalignment from the purely manual to an assisted activity of the surgeon (automation). Automation defines a system, that partly or totally fulfils function, those was carried out before totally or partly by the user. The organization of surgical assistance systems following application (planning, simulation, intraoperative navigation and visualization) or technical configuration of the system (manipulator, robot) is not suitable for a description of the interaction between user (surgeon) and the system. The available work has the goal of providing a classification for the degree of the automation of surgical interventions and describing by examples. The presented classification orients itself at pre-working from the Human-Factors-Sciences. As a condition for an automation of a surgical intervention applies that an assumption of a task, which was alone assigned so far to the surgeon takes place via the system. For both reference objects (humans and machine) the condition passively or actively comes into consideration. Besides can be classified according to which functions are taken over during a selected function division by humans and/or the surgical assistance system. Three functional areas were differentiated: "information acquisition and -analysis", "decision making and action planning" as well as "execution of the surgical action". From this results a classification of pre- and intraoperative surgical assist systems in six categories, which represent different automation degrees. The classification pattern is described and illustrated on the basis of surgical of examples. PMID- 17407010 TI - [Conflict of liability for medical malpractice and legal obligation for economic medical treatment]. PMID- 17407011 TI - [Non-syndromic hereditary hearing impairment]. AB - Hearing impairment is the most common sensorineural disorder in humans. Approximately one of thousand new-borns is affected by severe to profound deafness at birth or during early childhood. Genetic causes account for around half of these cases of prelingual hearing impairment and the remainder are attributed to environmental factors. Genetic causes of hearing impairment in combination with a syndrome as Usher, Pendred are distinguished from non syndromic hearing impairment. In the last years a tremendous growth in the localisation and identification of genes for non-syndromic hereditary hearing impairment has evolved. It has become clear that these conditions are genetically extremely heterogeneous. Approximately 120 different gene loci associated with non syndromic hearing impairment have been identified. Presently 54 gene loci associated with autosomal dominant mode of inheritance and 67 gene loci with autosomal recessive mode of inheritance have been identified; 7 are X-chromosome linked and 4 mitochondrial. Of these, 19 genes have been characterised for autosomal dominant (DFNA), 20 for autosomal recessive (DFNB), and 2 for X-linked (DFN) disorders. These genes encode proteins of diverse functions, including transcription factors, cytoskeletal and extracellular matrix components, and ion channels. Despite this heterogeneity, up to 50 % of prelingual recessive non syndromic deafness can be attributed to mutations in the GJB2 gene (Connexin-26, gap-junction protein). However, the diversity of genes and genetic loci implicated in hearing loss illustrates the complexity of the genetic basis of hearing. Knowing the gene and the function of its products helps understanding the mechanisms of hearing. PMID- 17407013 TI - Bleeding tendency as a first symptom in children with congenital biliary dilatation. AB - AIM OF THE STUDY: Although a bleeding tendency as a first symptom is a critical condition in congenital biliary dilatation (CBD), the clinical details of this symptom remain unclear. We assessed this condition in children with CBD in this paper. MATERIALS AND METHODS: Sixty-five children with CBD were treated at our institute between 1983 and 2004. The children, initially presenting with bleeding manifestations such as intracranial hemorrhage and bloody stools, were defined as the bleeding group, and the remaining children with digestive symptoms such as abdominal pain and vomiting were defined as the digestive group. The clinical features were compared between these two groups. RESULTS: In 6 of the 65 cases, bleeding manifestations were noted (9.2 %). All six had cystic-type choledochal dilatation. The mean age of the bleeding group was significantly younger than that of the digestive group, and bleeding was more frequent, especially in infants less than 12 months of age. In a laboratory study, the bleeding group showed a more prolonged blood coagulation time than the digestive group did. Serum amylase and lipase levels in the bleeding group were almost normal, while those in the digestive group were significantly higher. The direct bilirubin level in the bleeding group was significantly higher than that in the digestive group. CONCLUSIONS: Disturbed blood coagulation due to vitamin K deficiency related to cholestasis results in a bleeding tendency in children with CBD. Therefore, pediatric surgeons should be aware of this rare but critical condition which can be prevented by rapid and precise treatment with vitamin K supplementation. PMID- 17407014 TI - Comparison between prenatally diagnosed choledochal cyst and type-1 cystic biliary atresia by CD56-immunostaining using liver biopsy specimens. AB - Because it is difficult to distinguish preoperatively between a prenatally diagnosed choledochal cyst (CC) and type-1 cystic biliary atresia (BA) by ultrasound scanning or magnetic resonance imaging, some mode of discriminating between the 2 entities is required. The aim of this study was to investigate the immunohistological differences between prenatally diagnosed CC and type-1 cystic BA, using liver biopsy specimens immunostained for CD56. Five children with prenatally diagnosed CC and two children with prenatally diagnosed type-1 cystic BA were identified by fetal ultrasonography between 1985 and 2004. The control group included two children who were operated on at an earlier period due to postnatally diagnosed BA. Liver wedge biopsy in the right lobe was performed at the time of the radical operation. Histological findings of the CD56-stained liver biopsy specimens were classified into 4 categories each, with particular focus on staining distribution and intensity. The staining distribution was classified according to the scale 0 = no staining; 1 = some staining of bile ducts/ductules but staining in less than one-third of portal tracts; 2 = staining in one-third to less than two-thirds of portal tracts; and 3 = staining in more than two-thirds of portal tracts. Staining intensity was scored as follows: 0 = no staining, 1 = weak staining, 2 = moderate staining, and 3 = strong staining. The staining intensity and distribution in the CC group was zero in all 5 cases. The type-1 cystic BA group consisted of patients with scale 1 or 3 staining distribution and score 1 or 2 staining intensity. In the control group, staining distribution was 1 or 3, and staining intensity was 1 or 3. These results indicate that CD56-positive biliary duct cells are present in prenatally diagnosed type-1 cystic BA. The authors suggest that exploratory laparotomy might be avoided and, instead, immunohistological examination using liver biopsy specimens may be a reliable test for the differential diagnosis of CC and type-1 cystic BA in prenatally diagnosed neonates. PMID- 17407015 TI - Hepatic fibrosis with choledochal cyst in infants and children - an immunohistochemical assessment. AB - METHODS: The study population was divided into 4 groups: 12 infants with choledochal cyst, aged 4 m to 12 m, were classified as the infant choledochal cyst (ICC) group; 36 children, aged 1 y to 14 y, were classified as the children with choledochal cyst (CCC) group; while 18 patients, aged 2 m to 5 m, with biliary atresia (BA) were included as positive controls; and 14 infants, aged 1 d to 3 y, who died from non-liver diseases served as negative controls (CON). Liver specimens were examined using H&E sections to score fibrosis by means of Ohkuma's classification, and immunohistochemical sections were evaluated by counting the cells positive for cytokeratin (CK) and human leukocyte antigen-DR (HLA-DR) to discover the pathogenic factors of fibrosis. RESULTS: Most ICC patients had clinical biliary obstruction. The liver fibrosis score was highest in the BA group (2.9 +/- 0.7). The fibrosis score in the ICC group was higher (2.5 +/- 0.9) than that of the CCC (1.5 +/- 1.2; p < 0.05) and of the CON (0.1 +/- 0.4; p < 0.01) groups. The densities of CK-positive cells were 164 +/- 80/HP, 253 +/- 165/HP, 70 +/- 57/HP and 23 +/- 12/HP in the BA, ICC, CCC and CON groups, respectively, and differed significantly (p < 0.01) with the exception of the ICC vs. the BA group. The densities of HLA-DR positive cells were 130 +/- 72/HP, 98 +/- 54/HP, 96 +/- 50/HP and 36 +/- 13/HP in the portal area in the BA, ICC, CCC and CON groups, respectively. The density was lowest in the CON group (p < 0.01). CONCLUSION: In patients with choledochal cyst, liver fibrosis is more common and severe in infants than in children. Obstruction of the bile duct and proliferation of bile duct cells were the main pathogenic factors for fibrosis, while HLA-DR mediating immuno-injury may play a limited role. PMID- 17407016 TI - Late results of a modified new surgical procedure for the treatment of portal hypertension. AB - PURPOSE: The aim of this study is to report on the late results of the modified surgical procedure "subtotal splenectomy with omentopexy and partial splenic attachment to the parietal peritoneum" for the treatment of portal hypertension in children. METHODS: Over a period of twenty years (1984 - 2004), two modifications have been performed of the procedure of "partial splenectomy and partial splenic attachment for the treatment of portal hypertension", which were reported in the Zeitschrift fur Kinderchirurgie [ ], in 1990. The modifications were due to the recurrence of splenomegaly in five cases, with signs of hypersplenism in three of them after ten years. RESULTS: The follow-up ranges between one and fourteen years. All the patients are in good health, with good growth and development, except for one case with liver failure due to liver cirrhosis. No mortality occurred in this group of patients. Severe bleeding occurred in only one case after four years. Treatment consisted of sclerotherapy and ligation of the splenic artery. Splenomegaly was found in five cases, with hypersplenism in three of them. Splenic artery embolisation in two cases and ligation in one case produced good results. Howell-Jolly bodies were negative in all cases and radionuclide scan showed satisfactory to good results in most cases. Good results were also obtained in the studies of immunoglobulins and complements in most cases, with two exceptions of reduced IGM in one case and C4 in the other. CONCLUSIONS: Subtotal splenectomy with omentopexy and partial splenic attachment is a good alternative to other operative procedures for the treatment of portal hypertension. With this method, the precipitation and increase in collateral circulation leads to rapid improvement. PMID- 17407017 TI - Insights into infant neuroblastomas based on an analysis of neuroblastomas detected by mass screening at 6 months of age in Japan. AB - BACKGROUND/PURPOSE: Mass screening (MS) for neuroblastoma (NB) at 6 months of age in Japan was discontinued in 2004. We have previously reported that the majority of NB detected by MS showed a good prognosis, with only a few cases demonstrating an unfavorable outcome (J Pediatr Surg 2002, Cancer 2001). This study aims to provide insights into infant NB by assessing the details of the clinical courses in patients treated with a standard regimen and the biological features of such cases using highly sensitive methods at one institution in Japan. METHODS: In 76 NB detected through MS treated at Kyushu University Hospital, the clinical features and MYCN amplification, 1p deletion, 17q gain, the expression level of TRKA using FISH and the quantitative PCR were analyzed. RESULTS: Of these 76 persons with NB treated at one institution, 97 % are still alive, while 2 cases died from other diseases. Three patients experienced a recurrence after complete remission (CR), and 2 patients demonstrated refractory disease since the initial diagnosis. Two of the 3 NB patients with recurrence have demonstrated a 2nd CR, while one case still has multiple active diseases. Regarding the findings of highly sensitive biological analyses, 5/74 (7 %) showed MYCN amplification, 2/24 (8 %) cases had a 1p deletion, 3/33 (9 %) cases had a 17q gain, 5/50 (10 %) cases had diploidy, 1/25 (4 %) cases had a low expression of TRKA, and 2/76 (3 %) cases had an unfavorable histology. Of the 76 NB, 13 tumors (17 %) had one or more unfavorable factors (UF). Of the 5 refractory NB, 1 case had 3 UF, 1 case had 2 UF, 1 case had 1 UF, and 2 cases had no UF. As a result, 60 % of the refractory NB had one or more UF. CONCLUSIONS: Of the NB detected by MS at one institution in Japan, 17 % had one or more unfavorable factors (UF) and might have a higher risk of recurrence than the patients with no UF, although the unfavorable biology of several refractory cases is still unclear even after highly sensitive analyses. At least one-fifth of the NB cases detected by MS are anticipated cases. In infantile neuroblastomas, it may therefore be most important to analyze biologically prognostic factors using highly sensitive methods followed by immediate surgical intervention. Since the MS program has been discontinued in Japan, it will be necessary in future to assess the mortality and characteristics of NB detected clinically. PMID- 17407018 TI - Roux-en-Y feeding jejunostomy in infants and children. AB - Roux-en-Y jejunostomy (RYJ) permits enteral feeding in children unable to tolerate oral or intragastric feeds. It avoids many of the complications of nasojejunal and gastrojejunal tubes. Here we report our experience of intubated RYJ. By retrospective review of patient notes from 1998 - 2003, thirty-five children were identified. The median age was seven months (1 month - 16 years) and median follow-up was twenty-five months (1 - 55 months). There were no early postoperative complications or deaths. There were eighteen (51 %) late complications: peristomal leak (6), peristomal infection (2), bilious vomiting (5), tube displacement (3), abscess (1) and jejuno-colic fistula (1). Five children progressed to full oral feeds and had the RYJ resected. Fourteen of the twenty-eight children still alive remain fed by RYJ. Nine infants subsequently underwent fundoplication and gastrostomy with RYJ resection. Seven children died during the study period. RYJ is a straightforward procedure which can be performed safely in the face of poor nutrition and significant comorbidity. We recommend RYJ as a medium-term measure for enteral feeding. PMID- 17407019 TI - Neural crest neuroblasts can colonise aganglionic and ganglionic gut in vivo. AB - INTRODUCTION: Neural crest (NC) cells differentiate IN VITRO into neuroblasts, precursors of the enteric nervous system (ENS), when stimulated by specific agents. We developed a study aimed at establishing whether NC-derived neuroblasts can survive and colonise IN VIVO when injected into a recipient mouse gut. MATERIALS AND METHODS: The neuroblast precursors of the ENS were obtained from the vagal portion of the neural tubes of 296 CD-1 and GTROSA26 mouse embryos. The embryonic cells of GTROSA26 mice are identifiable through beta-galactosidase activity which allows recognition by blue staining. The host used in this study was the DOM/+ mouse, an animal model for Hirschsprung's disease (aganglionic megacolon). DOM/+ mouse pups (n = 43) received NC-derived cells inoculated into the seromuscular layer of the gut (33/43) or directly into the peritoneal abdominal cavity (10/43). RESULTS: All DOM/+ mice survived the procedure and were sacrificed after 7 or 14 days. Histochemical staining detected implanted cells in all mice. These showed specific myenteric colonisation into the aganglionic and ganglionic gut. CONCLUSION: The striking result of this study was the specific tropism of the injected NC-derived cells to target sites under the action of unknown chemotactic agents. This experimental procedure might represent a possible treatment option for specific forms of human ENS anomaly such as total intestinal aganglionosis. PMID- 17407020 TI - Experience and outcomes of nephroblastoma in Johannesburg, 1998 - 2003. AB - INTRODUCTION: Outcomes for children with cancer in the developing world are compromised by the difficulties for patients in accessing health services and by competition for resources between oncological services and the myriad other health problems of emerging nations. The purpose of this study is to document and analyse our experience and the outcomes of children with nephroblastoma over recent years. METHODS: This is a retrospective review of all patients who underwent combined oncological and surgical treatment for nephroblastoma in the Paediatric Oncology Unit between 1998 and 2003. RESULTS: Sixty-three patients were treated for Wilms' tumour; the mean age was 3 years 8 months (range 4 months to 11 years). The majority of children presented with an abdominal swelling or mass. Preoperative chemotherapy was given in forty-six cases (73 %). The tumour stage distribution was 11/63 stage I (17 %), 11/63 stage II (17 %), 21/63 stage III (33 %), 16/63 stage IV (25 %) and 4/63 stage V (6 %). Postoperative chemotherapy and radiotherapy was given according to the SIOP protocol. During the study period, thirteen patients (21 %) died (7 cancer-specific, 2 postoperative, 4 sepsis related), thirteen (21 %) were lost to follow-up and thirty-seven (59 %) are free of disease with a mean follow-up period of 3.67 years. Children with stage I and stage II had a disease-free survival at 4 years of 89 %. However, those with stage III, IV and V disease had 4-year survival of 66.75 % (p = 0.07). Overall, four-year post-nephrectomy survival was 76 %. CONCLUSION: Outcomes for children with cancer have improved dramatically over recent years; however, in the developing world, the scarcity of hospital resources and the overwhelming burden of non-cancer diseases can mean that oncological treatment is extremely challenging. In our society, children tend to present with nephroblastoma at an advanced stage; however, treatment by dedicated, multidisciplinary teams can achieve good results. PMID- 17407021 TI - Bone formation using human demineralised bone matrix (Grafton) for the treatment of bone cysts in children. AB - Bone cysts, in particular solitary bone cysts, are the most frequent cause of pathological fractures in children. However, there is still a great variety of regimens used to treat these lesions. Since demineralised bone matrix (DBM) is commercially available, we aimed to use this material for the consolidation of bones diagnosed as fragile because of cyst formation. Each of the 7 bone cysts as well as one enchondroma filled with DBM showed a continuous decrease in bone transparency over a period of two years (mean 8 months). A significant decrease in bone transparency and simultaneous cortical remodelling was radiographically detected in these cases as the specific hallmark of an initiated graft incorporation after 3 to 4 months. It was demonstrated that it is possible to heal children within an acceptable period of time using DBM to fill the cystic lesion. DBM appears to be a reasonable and beneficial alternative for the treatment of bone cysts offering both osteoinduction and osteoconductive features. PMID- 17407022 TI - Heterotopic rectal gastric mucosa: a rare cause of lower gastrointestinal bleeding in children. Case report and review of pediatric literature. AB - The authors report a very unusual case of intermittent and painful rectal bleeding occurring in a two-year-old female. At proctosigmoidoscopy, a bluish mucosal fold was found in the rectal ampulla. Histological examination showed the presence of heterotopic gastric mucosa islands intermingled with normal rectal mucosa. Surgical transanal excision of the lesion was undertaken due to the persistence of rectal bleeding with anemia, notwithstanding a three-month course of pharmacological therapy with a proton pump inhibitor. At a one-year follow-up, the child is well and symptom-free. The international pediatric literature was reviewed. PMID- 17407023 TI - Hydatid cyst presenting as a soft-tissue calf mass in a child. AB - Hydatid disease caused by Echinococcus granulosus often manifests as a slow growing cystic mass; it mainly affects the liver or lung and rarely other parts of the body such as the brain, heart, spleen, peritoneal cavity, or bone. Hydatid cysts of the musculoskeletal system are rare. Since the intramuscular hydatid cyst closely resembles a soft-tissue tumor on clinical examination, the preoperative radiological diagnosis is very important for the identification of the lesion prior to surgery. We describe a rare case of primary intramuscular hydatidosis in a child, together with its clinical presentation and radiological and histological findings. The treatment principles for hydatid cysts are also discussed. PMID- 17407024 TI - Traumatic rupture of hydatid cyst with unusual localization without liver involvement: a case report. AB - Primary hydatid disease of the intestinal mesentery is rare. We report on a 12 year-old male patient who presented with abdominal pain and urticaria lesions after blunt abdominal trauma. At laparotomy the cyst was found to be ruptured and was totally removed from the surrounding mesenteric tissue. The postoperative course was uneventful. This case is the first child reported with traumatic rupture of a hydatid cyst in a mesenteric location. PMID- 17407025 TI - Topical application of isosorbide dinitrate in patients with persistent constipation after pull-through surgery for Hirschsprung's disease. AB - AIMS: The aim of this study was to evaluate the anal manometric changes and the clinical effects after topical application of isosorbide dinitrate (ISDN) in patients with persistent constipation after pull-through surgery for Hirschsprung's disease (HD). METHODS: We studied 3 children (2 males and 1 female), aged 2, 3 and 5 years respectively, who had undergone the Soave-Boley surgical procedure for HD and who suffered from persistent constipation after operation. We performed a pre- and postoperative anorectal manometry study and we applied ISDN paste (1 mg/kg two times daily) in the anal region for three weeks. All patients were followed-up and re-evaluated at 1, 3, and 6 months. RESULTS: All patients showed an improvement of symptoms, with an average of 4 spontaneous evacuations per week. Prior to the topical treatment, the medium pressure was 115.6 mmHg (range 102 - 130 mmHg), the maximum pressure was 160 mmHg (range 145 - 175 mmHg), and the medium length of the high pressure zone was 1.8 cm (range 1.5 2.0 cm). At the 6 month follow-up, the medium pressure was 57.3 mmHg (range 52 - 61 mmHg, a decrease of 54.4 %), the maximum pressure was 98 mmHg (range 88 - 107 mmHg; a decrease of 38.7 %), and the medium length of the high pressure zone was 1.6 cm (range 1.4 - 1.8 cm; a decrease of 11.1 %). CONCLUSIONS: Topical treatment with ISDN is a valid therapeutic alternative to an anal myotomy in patients with persistent constipation after pull-through surgery for HD. However, a greater number of cases and a longer follow-up are necessary to confirm the validity of our experience. PMID- 17407026 TI - Conjoined epigastric heteropagus twins: excision of a parasitic twin from the anterior abdominal wall of her sibling. AB - Epigastric heteropagus twins (EHT) are an exceedingly rare form of asymmetric conjoined twins in whom the dependent twin (parasite) is attached to the right or left upper abdomen of the dominant part (autosite). Such a case observed at our institution with 34 month follow-up is presented here and the surgical technique described. A magnetic resonance imaging (MRI)-supported surgical separation of the parasite with successful closure of the abdominal wall defect of the autosite was performed. Follow-up studies showed an autosite which was alive and in optimal health. A comprehensive review including data from English and non English literature is presented. PMID- 17407027 TI - Sex differences in clinical leptospirosis in Germany: 1997-2005. AB - BACKGROUND: Although the majority of patients with reported leptospirosis are male, several seroepidemiologic studies demonstrated that leptospirosis seroprevalences are at least identical between both sexes. To study the effect of sex on the severity--and possibly, recognition--of leptospirosis, we conducted a sex-specific analysis of the clinical manifestations of the disease among patients with reported leptospirosis in Germany during 1997-2005. METHODS: Data on demographic characteristics, symptoms, place of infection and place of residence when infection occurred, possible exposure risks, infecting serovars, and mortality were evaluated for patients with laboratory-confirmed leptospirosis reported in Germany during 1997-2005. Mantel-Haenszel odds ratios (OR(MH)), adjusted for age, were used to determine the association between sex and the frequency of reported symptoms. RESULTS: Among 338 patients with leptospirosis (263 male patients and 75 female patients) reported in Germany during 1997-2005, male patients were more likely than female patients to be hospitalized (OR, 2.6; P<.01) and to exhibit symptoms of severe leptospirosis with jaundice (OR(MH), 3.7; P<.01), renal impairment (OR(MH), 3.4; P<.01), and hemorrhage (OR(MH), 7.8; P<.05). These sex-related differences were not associated with differences in exposure risks, presumptive infecting serogroups, or health-seeking behavior. CONCLUSIONS: Our results indicate that male sex is associated with a higher severity of clinical leptospirosis. Reports on male predominance in leptospirosis may thus reflect sex-related variability in the incidence of severe disease, rather than different infection rates. In clinical practice, sex differences in the manifestation of leptospirosis may cause systematic underinvestigation and undertreatment of the disease in female patients. PMID- 17407028 TI - Acute hepatitis with severe cholestasis and prolonged clinical course due to hepatitis A virus Ia and Ib coinfection. AB - BACKGROUND: Acute viral hepatitis due to hepatitis A virus is a self-limited illness that infrequently has a severe clinical course. METHODS: We analyzed the virological characteristics of acute hepatitis A in a patient with a severe clinical presentation (peak total and conjugated bilirubin levels, 65.5 mg/dL and 40.1 mg/dL, respectively) and a course of disease that lasted 7 months. RESULTS: Hepatitis A virus sequencing revealed coinfection with 2 subgenotypes of hepatitis A virus (Ia and Ib) as etiological factors of the illness. CONCLUSIONS: Hepatitis A virus Ia and Ib coinfection may have accounted for the prolonged and severe course of illness. PMID- 17407029 TI - An outbreak of Pneumocystis jiroveci pneumonia with 1 predominant genotype among renal transplant recipients: interhuman transmission or a common environmental source? AB - BACKGROUND: An outbreak of Pneumocystis jiroveci pneumonia (PCP) occurred among renal transplant recipients attending the outpatient department at the Leiden University Medical Centre (Leiden, The Netherlands) from 1 March 2005 through 1 February 2006. Clinical, epidemiological, and molecular data were analyzed to trace the outbreak's origin. METHODS: Renal transplant recipients with a clinical suspected diagnosis of PCP were included in the study. The diagnosis had to be confirmed by direct microscopy or real-time polymerase chain reaction of the dihydropteroate synthase gene in a bronchoalveolar fluid specimen. To detect contacts between patients, a transmission map was constructed. A case-control analysis was performed to asses whether infection was associated with certain wardrooms. Genotyping of Pneumocystis isolates was performed by sequence analysis of the internal transcribed spacer (ITS) number 1 and 2 gene regions. RESULTS: Twenty-two confirmed PCP cases were identified; approximately 0-1 would have been expected over the same time period. No risk factor was predominantly present, and standard immunosuppressive regimens had not changed. Liver transplant recipients who used the same outpatient facilities had not acquired PCP. The transmission map findings were compatible with interhuman transmission on multiple occasions. The case-control study did not point to wardrooms as a common source. Genotyping by sequencing of the ITS1 and ITS2 gene regions revealed type Ne in 12 of 16 successfully typed samples. Genotype Ne was found in only 2 of 12 reference samples. CONCLUSIONS: The clinical data and genotyping results are compatible with either interhuman transmission or an environmental source of infection. More complex models may account for PCP clusters. PMID- 17407030 TI - Transmission of Pneumocystis species among renal transplant recipients. PMID- 17407031 TI - An outbreak of severe respiratory tract infection due to human metapneumovirus in a long-term care facility. AB - BACKGROUND: Human metapneumovirus (hMPV) is a newly described paramyxovirus that is mainly associated with bronchiolitis in children. We sought to describe the epidemiological, virological, and histopathological findings associated with a large outbreak of hMPV infection in a long-term care facility. METHODS: An investigation of the outbreak was performed by public health authorities, who used standardized questionnaires to collect relevant clinical information from all residents of the facility. Nasopharyngeal samples were obtained from a subset of patients who had influenza-like illnesses for testing by viral culture and reverse-transcriptase polymerase chain reaction. Lung tissue samples from a patient whose case was fatal were available for molecular, histopathological, and immunohistochemical testing. RESULTS: A total of 96 (27%) of 364 residents of a long-term care facility presented with respiratory or constitutional symptoms between 1 January 2006 and 15 February 2006. The attack rate in the most affected ward was 72% (31 of 43 patients), which included 4 of the 6 polymerase chain reaction-confirmed cases of hMPV infection. In contrast, viral culture results were positive for hMPV in only 2 of the 5 polymerase chain reaction-positive samples tested. The most reported diagnosis was an upper respiratory tract infection or an influenza-like illness, although 21% of residents in 1 of the 3 wards that had confirmed cases of hMPV infection had lower respiratory tract infections. The fatality rate was 50% (3 of 6 patients) among confirmed cases and 9.4% (9 of 96 patients) among patients with possible cases. A patient with a fatal case had histopathological findings that confirmed the presence of hMPV RNA and proteins in the bronchiolar epithelium of affected lobes. Phylogenetic analysis revealed the presence of 2 distinct strains of hMPV circulating simultaneously on different wards. CONCLUSION: hMPV can be associated with important outbreaks of acute respiratory tract infection in elderly institutionalized persons. PMID- 17407032 TI - Respiratory tract infection due to human metapneumovirus among elderly patients. PMID- 17407033 TI - Coexistence of hepatitis B surface antigen (HBsAg) and heterologous subtype specific antibodies to HBsAg among patients with chronic hepatitis B virus infection. AB - BACKGROUND: The coexistence of hepatitis B surface antigen (HBsAg) and antibodies to HBsAg (anti-HBs) in patients with chronic hepatitis B virus (HBV) infection has been explained by the presence of viral escape mutants. Yet, no systematic analysis of such patients has been performed. We analyzed both the HBV strains and the nature of anti-HBs in such patients. METHODS: Four hundred eleven patients with chronic HBV infection were tested for the presence of anti-HBs. The sequences of the HBsAg coding region were analyzed. Anti-HBs were purified and examined in commercial assays alone and with 3 different HBsAg subtypes. RESULTS: Twenty patients had positive results for anti-HBs. This serological status remained stable for 12 months (as tested thus far). Amino acid substitutions and/or variations on HBsAg were found in 13 patients, and the HBV isolates from 4 others were wild types. Importantly, no significant difference in the occurrence of amino acid substitutions within the HBsAg was found in HBV isolates from patients with and without anti-HBs. Purified immunoglobulin fractions from serum samples from patients were reactive to HBsAg but had a lower specific activity, compared with those taken from immunized persons. Anti-HBs in patients were directed to the HBsAg subtypes other than the coexisting one. No circulating immune complex could be detected in these patients. CONCLUSION: HBsAg and anti HBs with an unmatched specificity coexisted in 4.9% of patients. The presence of anti-HBs was not associated with the appearance of specific HBV mutants in patients with chronic infection. Apparently, the presence of anti-HBs in patients with chronic HBV infection did not lead to a selection of HBV escape mutants. PMID- 17407034 TI - The enigma of concurrent hepatitis B surface antigen (HBsAg) and antibodies to HBsAg. PMID- 17407035 TI - Anticapsular serum antibody concentration and protection against pneumococcal colonization among children vaccinated with 7-valent pneumococcal conjugate vaccine. AB - BACKGROUND: Pneumococcal conjugate vaccines prevent invasive and noninvasive disease due to infection with vaccine serotypes. Pneumococcal conjugate vaccines also prevent nasopharyngeal acquisition of vaccine serotypes, although the mechanism is incompletely understood. METHODS: An efficacy trial of a 7-valent pneumococcal conjugate vaccine was conducted on the Navajo and White Mountain Apache reservations, located in the Southwestern United States; group C meningococcal conjugate vaccine was the control vaccine. Infants were randomized to receive 7-valent pneumococcal conjugate vaccine or group C meningococcal conjugate vaccine at 2, 4, 6, and 12 months of age. Immunogenicity and nasopharyngeal colonization studies were nested in the efficacy trial. We analyzed the correlation between serotype-specific serum IgG concentration at 7 and 13 months of age and nasopharyngeal acquisition of disease at 12 and 18 months of age, respectively. We adjusted for potential confounders using multivariate logistic regression. RESULTS: Among 203 subjects, we observed 60 acquisitions of vaccine-type pneumococci, including 19 acquisitions of serotype 19F (31.7%), and 17 acquisitions of serotype 23F (28.3%). Among recipients of 7 valent pneumococcal conjugate vaccine, increased serotype-specific serum IgG was associated with a reduction in nasopharyngeal acquisition of serotype 23F (relative risk, 0.53; 95% confidence interval, 0.31-0.93) but was not associated with a reduction in acquisition of serotype 19F (relative risk, 1.07; 95% confidence interval, 0.57-2.03). Among group C meningococcal conjugate vaccine recipients, serotype-specific serum IgG was not associated with a reduction in nasopharyngeal acquisition for either serotype. CONCLUSION: An increase in serum antibody concentration was associated with reduced acquisition of serotype 23F pneumococcus (but not with reduced acquisition of serotype 19F pneumococcus) among recipients of 7-valent pneumococcal conjugate vaccine. Differences in antibody concentration, in the functional characteristics of antibody, or in antibody kinetics during infancy may account for differences in carriage protection. PMID- 17407036 TI - Impact of prior antiplatelet therapy on risk of embolism in infective endocarditis. AB - BACKGROUND: Embolism is a dreaded complication of infective endocarditis (IE). Currently, antimicrobial therapy is the only medical intervention proven to decrease the risk of embolism associated with IE. We hypothesized that, because platelet aggregation is operative in the pathogenesis of vegetation formation, embolism associated with IE should occur less frequently among patients who have received prior, continuous daily antiplatelet therapy for noninfectious reasons. METHODS: We studied a retrospective cohort of adult patients with a diagnosis of IE who presented to the Mayo Clinic (Rochester, MN) during 1980-1998. The cohort was divided into 2 groups on the basis of whether they had received continuous daily antiplatelet therapy for at least 6 months prior to the time of hospitalization for IE. Antiplatelet therapy included aspirin, dipyridamole, clopidogrel, ticlopidine, or any of combination of these agents. The primary end point was a symptomatic embolic event that occurred prior to or during hospitalization. Multivariable logistic regression was used to assess the impact of continuous daily antiplatelet therapy on risk of symptomatic emboli associated with IE. RESULTS: One hundred forty-seven (24.5%) of 600 patients experienced a symptomatic embolic event; the most common embolic manifestation was stroke (in 48.2% of patients). Embolic events occurred significantly less often among those who had received prior, continuous daily antiplatelet therapy (12.0% of patients who had received therapy vs. 27.8% patients who had not receive therapy; P<.001). After adjustment for several covariates known to influence both risk of embolism and propensity for antiplatelet use, the adjusted odds ratio for a symptomatic embolic event was 0.36 (95% confidence interval, 0.19-0.68; P=.002) for patients receiving continuous daily antiplatelet therapy. CONCLUSIONS: The risk of symptomatic emboli associated with IE was reduced in patients who received continuous daily antiplatelet therapy before onset of IE. PMID- 17407037 TI - Outcomes attributable to neonatal candidiasis. AB - BACKGROUND: The incidence of candidiasis has increased in neonatal intensive care units, and invasive candidiasis is associated with significant morbidity and mortality. However, few data exist on outcomes directly attributable to neonatal candidiasis. METHODS: We estimated the incidence of systemic candidiasis in hospitalized neonates within the United States and determined the attributable mortality, length of hospital stay, and associated costs. We used the 2003 Kid's Inpatient Database from the Healthcare Cost and Utilization Project. Systemic candidiasis and comorbidities were defined by International Classification of Diseases, Ninth Revision, Clinical Modification codes. Neonates with uncomplicated births and neonates who died within the first 3 days of life were excluded. We used propensity score methods to balance covariates between the neonates with and neonates without candidiasis. Attributable outcomes were calculated between propensity score-matched neonates with and neonates without candidiasis. Because of the known confounding effect of birth weight, we performed separate propensity score analyses for extremely low birth weight (ELBW) neonates (i.e., neonates weighing < 1000 g). RESULTS: The overall incidence of invasive candidiasis in neonates is 15 cases per 10,000 neonatal admissions (95% confidence interval [CI], 13-16 cases per 10,000 neonatal admissions). ELBW neonates with invasive candidiasis were 2 times more likely to die (odds ratio, 2.2; 95% CI, 1.4-3.5) than propensity-matched ELBW neonates without candidiasis. The propensity score-adjusted mortality rate attributable to candidiasis among ELBW neonates was 11.9%. Candidiasis in ELBW infants was not associated with an increase in length of hospital stay but was associated with a mean increase in total charges of $39,045 (95% CI, $1374-$76,715). Among infants with a birth weight > or = 1000 g, those who had candidiasis did not experience a significant increase in mortality, compared with infants without candidiasis. However, the propensity score-adjusted length of stay and charges attributable to candidiasis among neonates with a birth weight > or = 1000 g were 16 days (95% CI, 8-24 days) and $122,302 (95% CI, $80,457-$164,148), respectively. CONCLUSIONS: Invasive candidiasis is associated with a significantly increased risk of death and excess hospital charges in ELBW neonates and with excess hospital stay and excess hospital charges in neonates with a birth weight > or = 1000 g. PMID- 17407038 TI - Diabetes mellitus and pyogenic liver abscess: risk and prognosis. AB - BACKGROUND: Pyogenic liver abscess (PLA) is a rare, life-threatening disease with an increasing rate of incidence. Case reports from East Asia suggest that diabetes mellitus is an important risk factor, but formal evidence is limited. METHODS: We performed a case-control study with participants drawn from the entire population of Denmark. Cases of PLA were defined as occurring in all patients who received a first-time diagnosis of PLA on hospital discharge between 1977 and 2002, as identified in the nationwide Danish National Patient Registry. Fifty sex- and age-matched population control subjects were selected for each patient with PLA. We computed the relative risk of PLA associated with diabetes using conditional logistic regression and controlling for major potential confounders. We further examined whether diabetes increased the relative risk of death until 30 days after hospital discharge among patients with PLA. RESULTS: We identified 1448 patients who experienced a first hospitalization for PLA during the study period (median age, 64 years; male sex, 54.2%). Persons with diabetes had a 3.6-fold increased risk of experiencing PLA, compared with population control subjects (adjusted relative risk, 3.6; 95% confidence interval, 2.9-4.5]. In addition, patients with PLA who had diabetes had a higher 30-day postdischarge mortality rate, compared with patients with PLA who did not have diabetes (24.8% vs. 18.0%). After controlling for other prognostic factors, the relative risk of death for patients with PLA and diabetes was 1.3 (95% confidence interval, 0.9 2.1). CONCLUSIONS: Diabetes is a strong, potentially modifiable risk factor for PLA. PLA is associated with a similarly poor prognosis for patients with diabetes and for other patients. PMID- 17407039 TI - The early history of coccidioidomycosis: 1892-1945. AB - Coccidioidomycosis was first discovered by a medical student in Argentina in 1892, and knowledge about the infection mostly arose from observations of clinicians and scientists in California, primarily at Stanford University Medical Center. Some discoveries came by chance. Many others arose from careful epidemiologic and clinical investigations in the San Joaquin Valley during the 1930s, when people migrated there from the "Dust Bowl" of the Midwest, and during the 1940s, when the events of World War II brought military recruits, prisoners of war, and persons of Japanese descent to camps and other areas of endemicity. Especially impressive were the contributions of Charles E. Smith, who tirelessly studied this disease throughout his professional career. PMID- 17407040 TI - The rationale for revising the Clinical and Laboratory Standards Institute vancomycin minimal inhibitory concentration interpretive criteria for Staphylococcus aureus. AB - The Clinical and Laboratory Standards Institute (formerly, the NCCLS) established the susceptibility and resistance breakpoints for minimal inhibitory concentration (MIC) and disk diffusion testing of vancomycin against isolates of Staphylococcus aureus > 20 years ago. The disk diffusion breakpoints were modified in 1998 when it was recognized that vancomycin-intermediate S. aureus strains were not detected by this method. In 2006, the vancomycin MIC breakpoints for S. aureus were lowered (from < or = 4 microg/mL to < or = 2 microg/mL for "susceptible," from 8-16 microg/mL to 4-8 microg/mL for "intermediate," and from > or = 32 microg/mL to > or = 16 microg/mL for "resistant") to increase detection of heterogeneously resistant isolates of S. aureus. This decision reflected a growing amount of microbiological and clinical data indicating that isolates of S. aureus are less likely to respond to vancomycin therapy when the vancomycin MICs are > or = 4 microg/mL. PMID- 17407041 TI - Evaluation of polymerase chain reaction and culture for diagnosis of pertussis in the control of a county-wide outbreak focused among adolescents and adults. AB - During a large pertussis outbreak, culture and polymerase chain reaction (PCR) were used to identify 149 case patients; of these case patients, 79 had positive PCR and culture results, 59 had positive PCR results and negative culture results, 11 had negative PCR results and positive culture results (10 PCR negative, culture-positive specimens were collected < or = 14 days after illness onset). PCR and culture of samples obtained < or = 2 weeks after illness onset and PCR of samples obtained > 2 weeks after illness onset proved to be most diagnostically useful. PMID- 17407042 TI - Cellulitis in a patient with leukemia. PMID- 17407043 TI - Syphilis and HIV infection: an update. AB - The striking increase in the prevalence of concordant human immunodeficiency virus (HIV) infection and syphilis observed by clinicians and public health officers over the past decade has renewed interest in the subject. Although the effect of HIV infection on the natural history of syphilis has been known for a long time, it was not until recently that several studies documented that syphilis may also impact the course of HIV infection. Despite an improved understanding of the interaction of these 2 conditions, many controversies still exist. In this article, we focus on the most recent literature describing the epidemiology, clinical manifestations, and treatment of syphilis in the context of HIV infection. PMID- 17407044 TI - Relationship between human T lymphotropic virus (HTLV) type 1/2 viral burden and clinical and treatment parameters among patients with HIV type 1 and HTLV-1/2 coinfection. AB - BACKGROUND: Human T lymphotropic virus types 1 (HTLV-1) and 2 (HTLV-2) are frequent copathogens among individuals infected with human immunodeficiency virus type 1 (HIV-1). The long-term effects of coinfection are unknown, and little information exists regarding how levels of HTLV-1/2 viral burden are affected by antiretroviral medications. METHODS: Factors associated with HTLV-1/2 viral burden were examined in patients with HIV-HTLV-1/2 coinfection. A total of 72 subjects were evaluated. The variables analyzed included HTLV-1/2 proviral load, HTLV-1/2 tax/rex mRNA expression, HIV load, HTLV-1/2 viral antigen detection in peripheral blood mononuclear cell (PBMC) cultures, T cell subsets, demographic variables (age, race, sex, and reported use of injection drugs), and administration of highly active antiretroviral therapy. RESULTS: An HTLV-1/2 proviral DNA copy number >20,000 copies/10(6) PBMCs was significantly associated with the following variables: (1) a positive HTLV-1 Western blot test result, (2) a positive HTLV-1/2 PBMC culture result, (3) a positive tax/rex mRNA result, (4) an HIV load <10,000 copies/mL, and (5) higher CD4 cell counts among subjects with HIV-HTLV-1 coinfection. There was no correlation between HTLV-1/2 proviral copy number or HTLV-1/2 tax/rex mRNA detection and administration of antiretroviral therapy. CONCLUSIONS: HTLV-1/2 proviral burden was significantly higher among patients with HIV-HTLV-1 coinfection than among patients with HIV-HTLV-2 coinfection. Highly active antiretroviral therapy may be of limited value in controlling virus expression of HTLV-1/2 in patients with HIV-HTLV-1/2 coinfection. PMID- 17407045 TI - Adherence to antiretroviral therapy and virologic suppression among HIV-infected persons receiving care in private clinics in Mumbai, India. AB - BACKGROUND: Adherence to antiretroviral therapy (ART) and correlates of adherence and virologic suppression among human immunodeficiency virus (HIV)-infected persons receiving ART in private, outpatient clinics in India is unknown. METHODS: Between December 2004 and April 2005, persons receiving ART at 3 private clinics in Mumbai, India, were interviewed regarding HIV care and adherence to ART. Physicians also completed a survey for each participant. Quantitative HIV-1 RNA level was determined for 200 participants. RESULTS: Of 279 participants, 73% reported > or = 95% adherence to ART. Adherence was positively associated with age > or = 50 years (adjusted odds ratio [aOR], 3.90), presence of comorbid conditions (aOR, 1.92), medication self-efficacy (aOR, 4.01), absence of pain in the past month (aOR, 2.14), and support from family and friends (aOR, 2.57). Lack of reminders from family members to take medication (aOR, 0.27) was negatively associated with adherence. Of 200 participants, 127 (63.5%) had virologic suppression (RNA level, < 400 copies/mL). Independent correlates of suppression were a regimen containing > or = 3 ART drugs (aOR, 5.52), first ART regimen (aOR, 3.28), adherence to therapy > or = 95% (aOR, 5.70), female sex (aOR, 3.19), and a physical component score > or = 50 (aOR, 1.07). CONCLUSION: Self-reported adherence to ART in a sample of patients attending Mumbai's private clinics was relatively high. However, the fact that a detectable viral level was found in nearly 40% of patients suggests that second-line ART regimens, as well as an emphasis on adherence and appropriate ART regimens in India, is needed. PMID- 17407046 TI - Treatment outcomes stratified by baseline immunological status among young children receiving nonnucleoside reverse-transcriptase inhibitor-based antiretroviral therapy in resource-limited settings. AB - A study of 568 children aged <5 years who commenced nonnucleoside reverse transcriptase inhibitor-based antiretroviral therapy in resource-limited settings revealed good early outcomes. After 12 months of antiretroviral therapy, survival probability was 0.89 (95% confidence interval, 0.86-0.92), with no significant difference among children stratified on the basis of baseline immunological levels; 62% attained a CD4 cell percentage >25%, and 7% continued to have a CD4 cell percentage <15%. PMID- 17407048 TI - Propensity-matched analysis of appropriate empirical antibiotic treatment. PMID- 17407049 TI - Difficulty in the rapid diagnosis of avian influenza A infection: Thailand experience. PMID- 17407050 TI - Rapidly fatal gas-forming pyogenic psoas abscess caused by Klebsiella pneumoniae. PMID- 17407051 TI - Emergence of fluoroquinolone only-resistant Escherichia coli in a community hospital. PMID- 17407053 TI - Comparison of novel clinically applicable methodology for sensitive diagnostics of cartilage degeneration. AB - In order efficiently to target therapies intending to stop or reverse degenerative processes of articular cartilage, it would be crucial to diagnose osteoarthritis (OA) earlier and more sensitively than is possible with the existing clinical methods. Unfortunately, current clinical methods for OA diagnostics are insensitive for detecting the early degenerative changes, e.g., arising from collagen network damage or proteoglycan depletion. We have recently investigated several novel quantitative biophysical methods, including ultrasound indentation, quantitative ultrasound techniques and magnetic resonance imaging, for diagnosing the degenerative changes of articular cartilage, typical for OA. In this study, the combined results of these novel diagnostic methods were compared with histological (Mankin score, MS), compositional (proteoglycan, collagen and water content) and mechanical (dynamic and equilibrium moduli) reference measurements of the same bovine cartilage samples. Receiver operating characteristics (ROC) analysis was conducted to judge the diagnostic performance of each technique. Indentation and ultrasound techniques provided the most sensitive measures to differentiate samples of intact appearance (MS=0) from early (13) degeneration. Furthermore, these techniques were good predictors of tissue composition and mechanical properties. The specificity and sensitivity analyses revealed that the mechano-acoustic methods, when further developed for in vivo use, may provide more sensitive probes for OA diagnostics than the prevailing qualitative X-ray and arthroscopic techniques. Noninvasive quantitative MRI measurements showed slightly lower diagnostic performance than mechano-acoustic techniques. The compared methods could possibly also be used for the quantitative monitoring of success of cartilage repair. PMID- 17407054 TI - Biodiversity and the survival of autochthonous fermented products. PMID- 17407055 TI - Bioinformatic analysis of outer membrane proteome of Neisseria meningitidis and Neisseria lactamica. AB - Two-dimensional electrophoresis (isoelectric focusing/SDS-PAGE) and Western blotting techniques were used to analyze and compare common and/or specific outer membrane proteins and antigens from Neisseria meningitidis and Neisseria lactamica. Bioinformatic image analyses of proteome and immunoproteome maps indicated the presence of numerous proteins and several antigens shared by N. meningitidis and N. lactamica, although the inter-strain variation in the maps was of similar magnitude to the inter-species variation, and digital comparison of the maps did not reveal proteins found to be identical by MALDI-TOF fingerprinting analysis. PorA and RmpM, two relevant outer-membrane antigens, manifested as various spots at several different positions. While some of these were common to all the strains analyzed, others were exclusive to N. meningitidis and their electrophoretic mobilities were different than expected. One such spot, with a molecular mass of 19 kDa, may be the C-terminal fragment of RmpM (RmpM Cter). The results demonstrate that computer-driven analysis based exclusively on spot positions in the proteome or immunoproteome maps is not a reliable approach to predict the identity of proteins or antigens; rather, other identification techniques are necessary to obtain accurate comparisons. PMID- 17407056 TI - Influence of age of aggregates and prokaryotic abundance on glucose and leucine uptake by heterotrophic marine prokaryotes. AB - The kinetics of glucose and leucine uptake in attached and free-living prokaryotes in two types of microcosms with different nutrient qualities were compared. Microcosm type M1, derived from unaltered seawater, and microcosm type M2, from phytoplankton cultures, clearly expressed different kinetic parameters (Vmax/cell and K' m). In aggregates with low cell densities (M1 microcosm), the attached prokaryotes benefited from attachment as reflected in the higher potential uptake rates, while in aggregates with high cell densities (M2 microcosm) differences in the potential uptake rates of attached and free-living prokaryotes were not evident. The aging process and the chemical changes in aggregates of M2 microcosms were followed for 15-20 days. The results showed that as the aggregates aged and prokaryotic abundance increased, attached prokaryotes decreased their potential uptake rate and their K' m for substrate. This suggests an adaptive response by attached prokaryotes when aggregates undergo quantitative and qualitative impoverishment. PMID- 17407057 TI - Microscopic and transcriptome analyses of early colonization of tomato roots by Trichoderma harzianum. AB - The capacity of the fungus Trichoderma harzianum CECT 2413 to colonize roots and stimulate plant growth was analyzed. Tobacco seedlings (Nicotiana benthamiana) transferred to Petri dishes inoculated with T. harzianum conidia showed increased plant fresh weight (140%) and foliar area (300%), as well as the proliferation of secondary roots (300%) and true leaves (140%). The interaction between strain CECT 2413 and the tomato-root system was also studied during the early stages of root colonization by the fungus. When T. harzianum conidia were inoculated into the liquid medium of hydroponically grown tomato plants (Lycopersicum esculentum), profuse adhesion of hyphae to the plant roots as well as colonization of the root epidermis and cortex were observed. Confocal microscopy of a T. harzianum transformant that expressed the green fluorescent protein (GFP) revealed intercellular hyphal growth and the formation of plant-induced papilla like hyphal tips. Analysis of the T. harzianum-tomato interaction in soil indicated that the contact between T. harzianum and the roots persisted over a long period of time. This interaction was characterized by the presence of yeast like cells, a novel and previously undescribed developmental change. To study the molecular mechanism underlying fungal ability to colonize the tomato-root system, the T. harzianum transcriptome was analyzed during the early stages of the plant fungus interaction. The expression of fungal genes related to redox reactions, lipid metabolism, detoxification, and sugar or amino-acid transport increased when T. harzianum colonized tomato roots. These observations are similar to those regarding the interactions of mycorrhiza and pathogenic fungi with plants. PMID- 17407058 TI - Characterization of a sulfide-oxidizing biofilm developed in a packed-column reactor. AB - The potential of microbial mats to develop sulfide-oxidizing biofims was explored. A bioreactor specially designed for the treatment of sulfide-containing effluents was inoculated with a microbial-mat sample, and a complex microbial biofilm with sulfide-oxidation activity developed. The microbial composition of the biofilm was studied by pigment, microscopy, and 16S rRNA gene analyses. Purple sulfur bacteria and diatoms were observed by microscopy, chlorophyll a and bacteriochlorophyll a were detected in the pigment analysis, and high genetic diversity was found in the 16S rRNA gene library. Specialized anaerobic sulfur oxidizers (i.e., phototrophic purple and green sulfur bacteria) dominated the library. Aerobic phototrophs (diatoms) also developed and the oxygen produced allowed the growth of aerobic sulfide oxidizers, such as Thiomicrospira-like spp. Cyanobacteria, which are significant organisms in natural microbial mats, did not develop in the reactor but unexpected uncultured members from the Epsilonproteobacteria developed profusely. Moreover, a variety of more minor organisms, such as members of the Cytophaga-Flavobacterium-Bacteroides (CFB) and purple non-sulfur bacteria (Roseospirillum sp.), were also present. The results showed that a complex community with high genetic and metabolic diversity, including many uncultured organisms, can develop in a laboratory-scale reactor. PMID- 17407059 TI - UV-radiation-induced formation of DNA bipyrimidine photoproducts in Bacillus subtilis endospores and their repair during germination. AB - The spore photoproduct (SP) is the main DNA lesion after UV-C irradiation, and its repair is crucial for the resistance of spores to UV. The aims of the present study were to assess the formation and repair of bipyrimidine photoproducts in spore DNA of various Bacillus subtilis strains using a sensitive HPLC tandem mass spectrometry assay. Strains deficient in nucleotide excision repair, spore photoproduct lyase, homologous recombination (recA), and with wild-type repair capability were investigated. Additionally, one strain deficient in the formation of major small, acid-soluble spore proteins (SASPs) was tested. In all SASP wild type strains, UV-C irradiation generated almost exclusively SP (>95 %) but also a few by-photoproducts. In the major SASP-deficient strain, SP and by-photoproducts were generated in equal quantities. The status time of 60 min, >75% of the SP was repaired in wild-type strains and in the SASP-deficient strain, while half of the photoinduced SP was removed in the recA-deficient strain. SP-lyase-deficient spores repaired 20% of the SP produced. Thus, SP lyase, with respect to nucleotide excision repair, has a remarkable impact on the removal of SP upon spore germination. PMID- 17407061 TI - Detection of the aminoglycosidestreptothricin resistance gene cluster ant(6)-sat4 aph(3 ')-III in commensal viridans group streptococci. AB - High-level aminoglycoside resistance was assessed in 190 commensal erythromycin resistant alpha-hemolytic streptococcal strains. Of these, seven were also aminoglycoside-resistant: one Streptococcus mitis strain was resistant to high levels of kanamycin and carried the aph(3 ')-III gene, four S. mitis strains were resistant to high levels of streptomycin and lacked aminoglycoside-modifying enzymes, and two S. oralis strains that were resistant to high levels of kanamycin and streptomycin harbored both the aph(3 ')-III and the ant(6) genes. The two S. oralis strains also carried the ant(6)-sat4- aph(3 ' ')-III aminoglycoside-streptothricin resistance gene cluster, but it was not contained in a Tn5405-like structure. The presence of this resistance gene cluster in commensal streptococci suggests an exchange of resistance genes between these bacteria and enterococci or staphylococci. PMID- 17407060 TI - Necrotoxigenic Escherichia coli from sheep and goats produce a new type of cytotoxic necrotizing factor (CNF3) associated with the eae and ehxA genes. AB - Fecal samples from sheep and goats were screened by tissue-culture assays and PCR for the presence of necrotoxigenic Escherichia coli (NTEC) producing cytotoxic necrotizing factors (CNFs). Of the 18 NTEC strains assayed, four were positive for the cnf1 gene while 14 strains were negative for the cnf1 and cnf2 genes. All of the NTEC strains had the eae gene and most of them also carried the ehxA gene. Moreover, all the cnf1- cnf2- NTEC strains were negative for several virulence markers associated with CNF1+ or CNF2+ strains. The cnf gene present in one of these strains was sequenced and analysis of the gene product revealed a new type of CNF, which was named CNF3 (and the coding gene cnf3). Oligonucleotide primers were designed to PCR-amplify a fragment of cnf3. The results showed that all strains examined in this study, except one cnf1+strain, were cnf3+. The association of cnf3 with eae and ehxA suggests that cnf3+ NTEC strains might be pathogenic for humans. PMID- 17407062 TI - The recently proposed species Aeromonas sharmana sp. nov., isolate GPTSA-6T, is not a member of the genus Aeromonas. AB - A new species of the genus Aeromonas, Aeromonas sharmana sp. nov., was recently described on the basis of a single isolate, strain GPTSA-6T, obtained from a warm spring in India. The description of this new species included biochemical characterization, antibiotic susceptibility, cellular fatty-acid profiles, and 16S rRNA gene sequencing, but not DNADNA hybridization data. In the present article, phylogenetic analysis (branch distances in the tree and nucleotide signatures) of the 16S rRNA of isolate GPTSA-6T, together with certain phenotypic characteristics of the isolate reported in the earlier description, clearly indicate that this microorganism does not belong to the genus Aeromonas as known to date, although it falls within the radiation of the family Aeromonadaceae. Emendation from the List of Prokaryotic Names with Standing in Nomenclature is consequently proposed. PMID- 17407063 TI - The evolution of the Science Citation Index. PMID- 17407064 TI - [Germ line mutations in Chinese kindreds with von Hippel-Lindau syndrome]. AB - OBJECTIVE: To investigate the germ line mutations in Chinese kindreds with von Hippel-Lindau syndrome (VHL) and to explore its role in clinical management. METHODS: The clinical and familial data were reviewed from 6 Chinese kindreds with VHL, of which the VHL germ line mutation in 21 members was analysed by polymerase chain reaction and sequencing analysis. RESULTS: Among 6 kindreds, there were 5 type I and 1 type IIA. The germ line mutations consisted of 4 missense mutations, 1 nonsense mutation and 1 deletion, of which 4 mutations existed in exon 1, 1 in exon 2 and 1 in exon 3. Of the 21 members who volunteered for genetic analysis, 14 members presented the VHL germ line mutations, including 10 affected patients, 1 suspected patients and 3 carriers. CONCLUSION: The germ line mutations in Chinese kindreds with VHL could be dominant in exon 1. It plays an important role in early detection of asymptomatic patients and the carriers, in the diagnosis of VHL and the clinical screening for members in the VHL families. PMID- 17407065 TI - [Cloning and characterization of down-regulated genes in neural tube defects of golden hamster induced by hyperthermia]. AB - OBJECTIVE: To clone down-regulated genes in neural tube defects of golden hamster induced by hyperthermia and to explore the molecular mechanism. METHODS: A reversed subtractive cDNA library was constructed using suppression subtractive hybridization. Clones with inserts were selected by combination of alpha complementary phenomenon and colony PCR. Then sequence and homology analysis of the inserts were made. And mRNA expression conditions were confirmed by Northern blot. RESULTS: The reversed subtractive cDNA library was successfully constructed. Fifteen recombinant clones were sequenced and recognized homologous to known genes. The results of Northern blot showed that all these genes were down-regulated in defected neural tubes induced by hyperthermia compared to normal developed neural tubes. CONCLUSION: Some important genes are identified which might be involved in neural tube defects induced by hyperthermia. PMID- 17407066 TI - [Analysis of chromosome aberrations in the cell derived from primary cell culture of laryngeal carcinoma and the Hep-2 cell line]. AB - OBJECTIVE: To search for characteristic chromosome changes in primary laryngeal squamous cell carcinoma (LSCC) and Hep-2 cell line and to realize the relationship between the cytogenetic abnormality and the pathogenetic mechanism in LSCC. METHODS: The fresh resulted samples of LSCC were analyzed with an improved primary cell culture for chromosome preparation and G-banding technique. Hep-2 cell line was analyzed by high resolution banding technique. Molecular cytogenetics analysis was made by chromosome 6 painting probe. RESULTS: Four primary LSCC succeeded in primary cell culture and obtained metaphases, one was tetraploid, the other three were triploid. The chromosome mode of Hep-2 cell line was from 68 to 75 and fifteen marker chromosomes were found. The most structural abnormalities of chromosome in primary LSCC and HEP-2 cell line were unbalance translocation, terminal deletion and isochromosome. The complicate aberration in chromosome 6 was common in LSCC and Hep-2. CONCLUSION: 6q-, I(5p), 17p-, 5q- are considered as characteristic chomosome changs in LSCC. Fluorescence in situ hybridization (FISH) may enhance the ability of detecting complicated chromosome rearrangements and marker chromosomes, which could provide more value data to verify the chromosome characteristic aberration in LSCC. PMID- 17407067 TI - [A novel mutation of the alpha-L-iduronidase gene in a patient with mucopolysaccharidosis type I]. AB - OBJECTIVE: To investigate the molecular genetic mechanism of a Chinese patient with mucopolysaccharidosis type I (MPS I). METHODS: PCR-sequencing analysis was applied to detect the mutations in exons in alpha-L-iduronidase gene (IDUA) of the patient. Restriction fragment length polymorphism (RFLP) and allele-specific oligonucleotide hybridization (ASO) were used to confirm the identified mutations. PCR amplified DNA samples from 50 normal individuals were sequenced to demonstrate that the newly identified mutation was not polymorphism. RESULTS: The patient was compound heterozygous for a previously reported nonsense mutation Q60X (178C > T) in exon 2, inherited from the mother, and a newly detected missense mutation D203N (607G > A) in exon 6 from the father. The newly identified mutation D203N was not found in PCR amplified products from 50 normal individuals, indicating that it was not polymorphism. CONCLUSION: The two identified mutations may be the cause resulting in patient's clinical phenotype. PMID- 17407068 TI - [The clinical application of whole chromosome painting probes in preimplantation genetic diagnosis for translocation carriers]. AB - OBJECTIVE: To make preimplantation genetic diagnosis (PGD) for female translocation carriers by analyzing first polar bodies (1PBs) with whole chromosome painting probe (WCP). METHODS: WCP was used in fluorescence in situ hybridization (FISH) analysis of 1PBs for four female Robertsonian carriers presented for PGD with 45 XX, der(13;14)(q10;q10) karyotype. All the patients underwent ovarian stimulation and during 6 h after oocyte retrieval 1PBs were biopsied and WCP were used in FISH. On day 3 after fertilization embryos diagnosed as normal or balanced were transferred. RESULTS: A total of 61 oocytes were collected in 4 PGD cycles. Of the 54 matured oocytes, 50 were biopsied and 45 were fixed successfully. Results were obtained in 40 1PBs. Overall, 74.1% (40/54) oocytes were diagnosed. The fertilization rate and good embryo rate were 64.8% (35/54) and 65.7% (23/35) respectively. Two clinical pregnancies were obtained. One patient delivered a normal female baby with karyotype 46, XX in June 2006. For another patient, the fetus spontaneously aborted at 9th week of pregnancy with karyotype of 45, X confirmed by amniotic villus diagnosis. CONCLUSION: WCP can differentiate normal, balanced and unbalanced oocytes accurately and can be used as an efficient PGD method for female carriers of translocation. PMID- 17407069 TI - [The study of the correlation between the clinical phenotype of spinal muscular atrophy and SMN2 gene copy number]. AB - OBJECTIVE: To detect the correlation between the clinical phenotype of spinal muscular atrophy (SMA) and survival motor neuron gene (SMN2) copy number. METHODS: The SMN2 gene copy numbers of 57 different types of SMA were detected by real-time fluorescence quantitative PCR method with TaqMan technique. RESULTS: Average SMN2 copy number was 1.017 +/- 0.090, 2.019+/- 0.080, 3.104+/- 0.170 in predicting one, two, three copy numbers, respectively, and CV was 8.9%, 3.9%, 5.4%, respectively. Average SMN2 copy number was 1.926+/- 0.460, 2.508+/- 0.460, 2.876+/- 0.270, in type I, II and III SMA, respectively. The SMN2 gene copy number in type II and III SMA were higher than that of type I SMA (P < 0.01). The SMN2 gene copy number in type III SMA was higher than that of type II SMA (P < 0.01). 85.72% of type I SMA patients usually had 2 SMN2 copies; 40% and 60% of type II SMA patients had 2 and 3 SMN2 copies, respectively; 82% of type III SMA patients had 3 SMN2 copies. CONCLUSION: There is significant correlation between the change of SMA clinical phenotype and SMN2 cope number. The distributions of the SMN2 gene copy number are various in different types of SMA patients. All types of SMA patients have at least one copy SMN2. The SMN2 gene copy numbers in type II, III SMA are higher than that of type I. All of these findings suggest that the severity of SMA patients depend on the change of the SMN2 copy numbers. PMID- 17407070 TI - [A study on selective killing effect of Hsv-tk/GCV driven by human telomerase catalytic subunit promoter on human lung cancer cell A549]. AB - OBJECTIVE: To study selective killing effect of herpes simplex virus-thymidine kinase/ganciclovir (Hsv-tk/GCV) driven by human telomerase catalytic subunit (hTERT) promoter on lung cancer cell line A549 in vitro. METHODS: (1) Expression plasmids of Hsv-tk gene driven by hTERT promoter and sv40 promoter respectively (pGL3-hTp-tk and pGL3-sv40-tk) were transfected into telomerase-positive human lung adenocarcinoma cell line A549 and telomerase-negative fetal lung fibroblast cell line MRC-5. Reverse transcription-PCR was performed to detect expression of tk gene in above transfected cell lines; (2) Inhibition effect on proliferation of above transfected cell lines treated with GCV was investigated with MTT method; (3) Influence of GCV on apoptosis and cell cycle of above transfected cell lines was investigated with flow cytometry. RESULTS: (1) tk mRNA expression was detected in both A549 and MRC-5 transfected with pGL3-sv40-tk, also in A549 transfected with pGL3-hTp-tk, but not in pGL3-hTp-tk transfected MRC-5; (2) GCV showed significant inhibition effects on proliferation of pGL3-sv40-tk transfected A549 and MRC-5 in vitro, also on that of pGL3-hTp-tk transfected A549, but not on that of pGL3-hTp-tk transfected MRC-5; (3) Treated with GCV, apoptosis index (AI) of pGL3-sv40-tk transfected A549 and MRC-5 as well as pGL3 hTp-tk transfected A549 (21.58%, 9.35% and 23.19% respectively) increased significantly, compared with A549, MRC-5 transfected with pGL3-hTp (0.78% and 0.55% respectively) and A549, MRC-5 without plasmid transfection as blank control (2.17% and 0.60% respectively); GCV had no influence on AI of pGL3-hTp-tk transfected MRC-5 (0.88%). CONCLUSION: tk gene driven by hTERT promoter could express selectively in lung cancer cell A549. Hsv-tk/GCV driven by hTERT promoter could selectively inhibit proliferation of lung cancer cell. PMID- 17407071 TI - [Association of aldosterone synthase gene -344 T/C polymorphism with early renal damage in Han nationality with essential hypertension]. AB - OBJECTIVE: To investigate whether the -344T/C polymorphism of aldosterone synthase gene is associated with early renal damage in Han nationality with essential hypertension in Shandong province. METHODS: Plasma aldosterone concentration and urinary albumin excretion were measured with radioimmunoassays in 225 patients with essential hypertension, and hypertensives were classified as hypertension with normal albuminuria or hypertension with microalbuminuria according to urinary albumin excretion during 24 hours. -344T/C polymorphism of aldosterone synthase gene was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in controls and hypertensives. RESULTS: No significant differences were found in genotype distribution among groups of control, primary hypertension with normal albuminuria and hypertension with microalbuminuria. The C allele frequency in hypertension with microal buminuria group was significantly higher than that in control and hypertension with normal albuminuria group (P < 0.05). In hypertensive patients, plasma aldosterone concentration and urinary albumin excretion of TC+CC genotypes were significantly higher than that of TT genotype ( P< 0.05). CONCLUSION: These results suggest that -344T/C polymorphism of aldosterone synthase gene may be associated with early renal damage in Han nationality with essential hypertension, C allele may be a genetic factor susceptible to renal damage in hypertensives. PMID- 17407072 TI - [The genetic and clinical characteristics of transcription factor 1 gene mutations in Chinese diabetes]. AB - OBJECTIVE: To investigate the genetic and clinical features of mutations and sequence variations of the transcription factor 1 gene (TCF1, HNF-1A) in Chinese with familial early-onset and/or multiplex diabetes mellitus. METHODS: All ten exons of the TCF1 gene were screened, including exon and intron junctions, by direct sequencing method in 341 unrelated Chinese subjects, including 80 healthy controls and 261 probands of early-onset and/or multiplex diabetes pedigrees. RESULTS: Five mutations were found in all. Four of the 5 different TCF1 mutations were newly identified novel mutations(T82M, Q130H, G253G, P353fsdelACGGGCCTGGAGC), mean body mass index of mutation carriers was 21.9 kg/m (2), and insulin secretion was impaired in the mutation carriers. In this study, the maturity-onset diabetes of the young type III (MODY3) only accounted for 3% of Chinese early-onset diabetes. Moreover, eleven substitutions were identified in 261 probands. Of them, three variants IVS1-8 (G-->A), IVS1 -128 (T-->G ) and IVS2+21 (G-->A) were not observed in 80 healthy controls and one of them IVS1-8 (G-->A) was not reported previously and the two promoter variants co-segregated with diabetes. CONCLUSION: TCF1 gene is not a common cause of early-onset and/or multiplex diabetes among Chinese patients. PMID- 17407073 TI - [The role of insulin-like growth factor-2 gene differentially methylated regions in TCDD-induced malformation]. AB - OBJECTIVE: To study the effects of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) on the development of fetal rats and to explore the relationship between TCDD induced abnormal development in rats and the expression and the methylation of insulin-like growth factor 2 gene (Igf2). METHODS: A single dose of 10 microg/kg TCDD was given to gestation day (GD) 10 pregnant rats by gavage. On GD20, the fetuses were taken out and examined. The crown-rump length, the body weight and the placental weight were measured. The expression of Igf2 in liver was detected by real-time quantitative reverse transcription (RT-PCR) and Western blot. The methylation of Igf2 differentially methylated regions (DMRs) in liver was analyzed by a methylation-sensitive restriction enzyme Hpa II PCR assay and a bisulfite-modified DNA sequencing procedure. RESULTS: In the treatment group, 12.2% of the fetuses were either dead or absorbed, and 11.6% of them were malformed. For the live fetuses, their crown-rump length, body weight and placental weight were significantly lower than those of the control group. The relative amount of Igf2 mRNA in the treated livers and the control livers was 0.77 +/- 0.11 and 0.27+/- 0.15, respectively. The number was significantly higher in the treatment group than in the control group (P < 0.01). Western blot also showed a remarkable up regulation of Igf2 protein in liver after treatment. The two groups showed no difference in the methylation status of Igf2 DMR1 in liver. The DMR2 Igf2 was significantly hypomethylated in the treated livers than in the control livers. CONCLUSION: Exposure to TCDD in pregnancy can lead fetal rats to death, absorption, malformation and intrauterine growth retardation (IUGR). The TCDD led abnormal development in rats may be associated with the hypomethylated DMR2 of Igf2 and the up regulation of Igf2 in liver. PMID- 17407074 TI - [Study on the mitochondrial DNA variation in patients with type 2 diabetes mellitus]. AB - OBJECTIVE: To explore the relationship between type 2 diabetes mellitus (T2DM) and the mutations in the fragment of mitochondrial DNA (mtDNA) from nucleotides 3153 to 3551, which have shown high frequency of point mutation. METHODS: One hundred and ninety-one normal controls and 222 patients with T2DM were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), T-A cloning sequencing and denatured high performance liquid chromatography (DHPLC) techniques. RESULTS: The prevalence of mtDNA mutations in the patient group (24.32%) was significantly higher than that in the control group (7.33%) (P < 0.05). Three novel mutations of A3209T, T3253G and A3467C were found, and C3497T was first reported in DM. Onset age, body mass index, fasting blood glucose, HbA1C, high density lipoprotein-cholesterol and diabetic nephropathy could be related to occurrence of mtDNA mutations (P < 0.05). CONCLUSION: Mitochondrial DNA mutations might implicate T2DM in Wenzhou population, which should play an important role in the pathogenesis of T2DM. PMID- 17407075 TI - [Association of L-selectin gene polymorphism with susceptibility to coronary heart disease]. AB - OBJECTIVE: To investigate the possible association between L-selectin gene P213S polymorphism and coronary heart disease (CHD) in Chinese population. METHODS: In total 212 CHD patients diagnosed by angiography and 230 healthy controls were studied. The genotype and allele frequencies of L-selectin gene polymorphism were assayed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The frequency of the L-selectin gene 213P allele in CHD patients was significantly higher than that in the control group (77.59% vs 69.35%, P=0.006). Compared with the SS genotype, PP homozygote had a significantly increased CHD risk (OR=2.70 and OR=2.15 using unadjusted and adjusted Logistic regression models, respectively). No association was found between the severity of CHD and the Lselectin gene P213S polymorphism CONCLUSION: Our findings suggest that L-selectin gene 213P mutant allele might contribute to susceptibility of Chinese individuals to contract CHD. PMID- 17407076 TI - [Analysis of ATP binding cassette A1 gene R219K polymorphism in patients with endogenous hypertriglyceridemia in Chinese population]. AB - OBJECTIVE: To investigate the variations of ATP binding cassette A1 (ABCA1) gene and its relation to endogenous hypertriglyceridemia (HTG) in Chinese population. METHODS: A total of three hundred and nine subjects (one hundred and nine endogenous hypertriglyceridemics and 200 healthy controls) from a population of Chinese Han nationality in Chengdu area were studied using restriction fragment length polymorphism (RFLP) amplified by polymerase chain reaction (PCR). RESULTS: The frequency of K allele at R219K site 0.472 and 0.436 in normal and HTG group, respectively. No significant difference between normal control and HTG group was found in both allele and genotype frequencies. In both normal and HTG groups, subjects with genotype KK had a higher serum mean concentration of high density lipoprotein-cholesterol (HDL-C) when compared with those with genotype RR, respectively (1.48+/-0.45 mmol/L vs 1.27+/-0.29 mmol/L, P<0.05; 1.07+/-0.30 mmol/L vs 0.87+/-0.19 mmol/L, P<0.05). In normal group, subjects with genotype RK had a lower triglyceride (TG) level compared with those with genotype RR (1.22+/ 0.37 mmol/L vs 1.41+/-0.84 mmol/L, P<0.05). In addition, the subjects carrying K allele in HTG group had a decreased total cholesterol (TC)/HDL-C ratio compared with those with genotype RR (KK vs RK vs RR: 4.82+/-1.28 vs 5.42+/-1.62 vs 6.33+/ 1.70, P<0.05). CONCLUSION: These results suggest that R219K polymorphism in ABCA1 gene is not only associated with serum HDL-C and TG levels in healthy Chinese subjects in Chengdu area, but also with HDL-C level and TC/HDL-C ratio in subjects with endogenous HTG. PMID- 17407077 TI - [Identification of novel genes related to glioma by oligonucleotide microarray]. AB - OBJECTIVE: To identify the genes differentially expressed in development of human glioma, and to study the expression of some genes in different grade gliomas. METHODS: Oligonucleotide microarray (including 218 genes related to neural system development) was adopted and hybridized with probes which were prepared from the total RNAs of glioma specimens and normal brain tissues. Differentially expressed genes between the normal tissues and glioma tissues were assayed after scanning oligonuceltide microarray with ScanArray 4000, and some of these genes such as smad1, Hmp19 and TRIP3 were verified by real-time quantitative PCR(real-time-Q PCR) method. RESULTS: In comparison with the genes in the normal brain tissue, 5 down-regulated and 5 up-regulated genes in glioma specimens were revealed by means of microarrays, and the expression of smad1, Hmp19 and TRIP3 were verified by real-time-Q-PCR assay. CONCLUSION: Multiple genes play important roles in development of glioma. cDNA microarray technology is a powerful technique in screening for differentially expressed genes between glioma tissues and normal brain tissues. This study is helpful for judgement of invasion and prognosis of gliomas, and provides more target genes for targeted therapy. PMID- 17407078 TI - [Analysis of Kell blood group system using polymerase chain reaction-restriction fragment-single strand conformation polymorphism combined with heteroduplex in Chinese]. AB - OBJECTIVE: To investigate the polymorphism of Kell blood group system in Chinese and to find a suitable method for large scale screening. METHODS: An analysis method of polymerase chain reaction-restriction fragment-single strand conformation polymorphism (PCR-RF-SSCP) combined with heteroduplex was established to detect abnormal sample in KEL exon 7-9 area, then sequencing was used to find out the mutation site. RESULTS: Two mutations were found from 500 samples: 966G > A mutation in exon 9 and C > A mutation in 67th site of intron 7, both with no amino acid change. The mutation rate was 4/1000. No mutation was found as missed in using PCR-RF-SSCP combined with heteroduplex. CONCLUSION: PCR RF-SSCP combined with heteroduplex is confirmed as an effective, economical and simple method, it is quite suitable for large scale population screening study with unclear gene background and unavailable positive controls. Since there is special polymorphism for Kell blood group system in Chinese, further study is needed. PMID- 17407079 TI - [Loss of heterozygosity in the region of chromosome 4p15.1-4q12 in nasopharyngeal carcinoma patients with familial history]. AB - OBJECTIVE: To analyze the allelic loss of heterozygosity (LOH) in the region of chromosome 4p15.1-4q12 in nasopharyngeal carcinoma (NPC) patients with family history. METHODS: Tumor cells and lymphocytes were obtained from paraffin embedded biopsied tissue section by microdissection. LOH detections were carried out on 25 NPC patients with family history by PCR-based microsatellite polymorphism analysis using 7 pairs of microsatellite markers primers. The microsatellite loci located in 4p15.1-4q12 region. Genescan software was used to analyse LOH at each locus. RESULTS: Ninety-two percent of NPC cases (23/25) with family history was showed at least one microsatellite marker of LOH. Higher frequencies of LOH were found at three loci: D4S238 (56%), D4S350 (50%), D4S1547 (50%). The minimal common region of deletion might be defined between D4S350 and D4S1547. CONCLUSION: The higher incidence of LOH at D4S350 and D4S1547 suggests that there may be a potential tumor suppressor gene located in the two regions. PMID- 17407080 TI - [Detection of alpha thalassemia using real-time PCR and dissociation curve analysis]. AB - OBJECTIVE: To establish an automatic, high throughput, quick detection method of alpha thalassemia. METHODS: The genotypes of -alpha(4.2) and -alpha(3.7) were detected by two real-time fluorescence PCRs using SYBR-Green1 (SYBR-PCR) with the analysis of dissociation curve (DC) and melting temperature (Tm). The PCR products were recombined into T-vector and the correct cloning was selected as positive control. Sensitivities were gained from a serious dilution of the recombinant as SYBR-PCR template, from which detection deadlines for two kinds of alleles could be determined. Totally 110 samples were detected by this technique. RESULTS: The length of product of -alpha(4.2) and -alpha(3.7) were 1.65 kb and 1.9 kb respectively, and the Tm were (81.5+/-0.5) degrees C and (82.5+/-0.5) degrees C respectively. The detection deadline were 9 x 10(2 ) copies and 4.3 x 10(2) copies respectively. The sensitivity of the technique was much higher than that of regular PCR plus gel electrophoresis method, and the detection results of the technique were the same as that of multiplex PCR. CONCLUSION: The genotypes of -alpha(4.2), -alpha(3.7), non-deletion alpha alpha/and --(SEA) can be sensitively, exactly diagnosed by the real-time PCR with SYBR-Green1 combined with the dissociation curve analysis. The assay is automatic, low-cost, high throughput, and easy for quality control without fluorescence probe. PMID- 17407081 TI - [Non-invasive prenatal genetic diagnosis using multiple displacement amplification]. AB - OBJECTIVE: To investigate the feasibility of multiple displacement amplification (MDA) to apply in the non-invasive prenatal genetic diagnosis of Duchenne muscular dystrophy (DMD). METHODS: Maternal blood was obtained from 20 pregnant women at 7 to 25 weeks of gestation. After the discontinuous density gradient centrifugation with Percoll, the fetal nucleated red blood cells (NRBCs) were stained with Kleihauer test. All positive NRBCs were collected by micromanipulator and then performed with MDA. Sex and short tandern repeat (STR) analysis were determind from a small aliquot of the reaction. The origin of NRBCs was verified and prenatal diagnosis of DMD was made at the same time. RESULTS: The product length of MDA was >15 kb, while primer extension preamplification (PEP) is only about 1 kb. We completed non-invasive prenatal genetic diagnosis of 6 fetus at high risk of DMD using MDA. The results were all coincident with amniotic fluid control. CONCLUSION: The MDA method which provides a highly uniform representation across the genome, representing the entire genome with minimal amplification bias, shows good application prospects. PMID- 17407082 TI - [Application of real-time fluorescence quantitative PCR accompanied with comparison of Delta CT for diagnosis of Down's syndrome from a single cell]. AB - OBJECTIVE: To evaluate the use of real-time fluorescence quantitative PCR (FQ PCR) accompanied with comparison of Delta CT as a method for diagnosis of Down's syndrome from a single cell. METHODS: Single lymphocyte was isolated from the peripheral or umbilical cord blood samples of 22 clinically diagnosed Down's syndrome patients or fetus and 40 normal controls by micromanipulation techniques. Primer extension preamplification (PEP) and real-time FQ-PCR were used to amplify the S100B and DCSR1 located on chromosome 21, and GAPDH located on chromosome 12. Two pairs of DeltaCT values were compared between the two groups. The ratios of S100B/GAPDH and DSCR1/GAPDH products were calculated in trisomy 21 group. RESULTS: The DeltaCT values of Down's syndrome patients were significantly lower than those of normal controls. The difference between the two groups was statistically significant (P<0.01). The ratios of S100B/GAPDH and DSCR1/GAPDH products for trisomy 21 were 1.891(1.563-2.287) and 1.840 (1.562 2.168), respectively. CONCLUSION: Real-time FQ-PCR is a reliabe method that may provide a new way for non-invasive prenatal diagnosis and preimplantation genetic diagnosis for Down's syndrome. PMID- 17407083 TI - [Study on the vertical transmission of Y chromosome microdeletions from father to son]. AB - OBJECTIVE: To analyze the Y chromosome microdeletions in the family of the infertile male and to study on the vertical transmission of Y chromosome microdeletions from father to son. METHODS: The peripheral blood of infertile patients' family male members was extracted and analyzed with modified multiplex PCR. The infertile family tree was drawn according to the results. RESULTS: Two cases in twelve investigated families had azoospermia factor (AZFc) microdeletion heredity. The others had no heredity. CONCLUSION: AZFc microdeletion of the Y chromosome can be transmitted to the male offspring naturally,and the same deletion can result in different phenotypes in different individuals. PMID- 17407084 TI - [Relationship between polymorphisms of interleukin 10 promoter and serum levels of lipoprotein in the Chinese Han population]. AB - OBJECTIVE: To study the relationship between polymorphisms of interleukin 10 (IL10QX) promoter and serum levels of lipoprotein in the healthy Chinese Han population. METHODS: PCR restriction fragment length polymorphism assay was used to detect the distribution of genotypes of IL10 -592,-819,-1082 in 200 healthy Chinese Han subjects. Serum levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and very low-density lipoprotein (VLDL) in all subjects were measured to analyze the relationship with the polymorphisms of IL10 promoter. RESULTS: Comparing with AA genotype, the group with GA genotype at IL10 promoter -1082 position had a significant elevation of serum HDL-C level [(1.514+/-0.501) mmol/L vs. (1.261+/-0.346) mmol/L, t=-2.225, P=0.028] and a lower serum TG level[(1.701+/-1.836) mmol/L vs. (0.981+/-0.314) mmol/L,Z=-2.096,P=0.036]. The TG, TC, HDL-C, LDL-C and VLDL levels did not show any statistically significant differences among different genotypes (CC, AA, CA) of the IL10 -592, as well as the genotypes (TT, TA, AA) ofIL10 -819 (P>0.05). CONCLUSION: The results suggest that in the Chinese Han population, the polymorphism at position -1082 in the promoter region of IL10 gene may be associated with the serum HDL-C level and TG level. PMID- 17407085 TI - [Screening for tetrahydrobiopterin metabolic disorders and related gene analysis among the patients with motor disturbance and mental retardation]. AB - OBJECTIVE: To study the incidence of various enzyme deficiency in tetrahydrobiopterin (BH4) metabolism and the related gene mutation among the patients with motor disturbance and mental retardation. METHODS: One hundred patients with unknown motor disturbance and mental retardation were referred to this study. All patients were performed by phenylalanine (Phe) and BH4 loading test, urinary pterin analysis and dihydropteridine reductase (DHPR) activity. Some patients received the dopa treatment for diagnosis of dopa-responsive dystonia (DRD). The analysis of GTP cyclohydrolase 1 gene (GCH1) mutation for DRD patients and the analysis of 6-pyruvoyl tetrahydropterin synthase (PTS) gene mutations for PTS deficient patients were done under the consent from their parents. RESULTS: Seventy of 100 patients had normal basic blood Phe levels, six (6%) patients were diagnosed as DRD. Thirty patients had hyperphenylalaninemia (HPA), eight (8%) were diagnosed as PTS deficiency and 22(22%) were diagnosed as phenylalanine hydroxylase (PAH) deficiency. All patients had normal DHPR activity. The mutation IVS5+3insT of GCH1 was found in 2 patients with DRD. Seven kinds of PTS mutations were found in 8 patients with PTS deficiency, and 75% of the mutations were 259C-->T,286G-->A and 155A-->G. CONCLUSION: Some patients with unknown motor disturbance and mental retardation may suffer from BH4 metabolism related diseases. Theses patients are necessary to be screened for such kind of diseases in order to confirm the diagnosis. PMID- 17407086 TI - [Study on relationship between polymorphism sites of TIM4 and allergic asthma in the population of Han nationality from Hubei province of China]. AB - OBJECTIVE: To investigate two single nucleotide polymorphism sites of T cell immunoglobulin domain and mucin domain protein-4 (TIM4) and to detect their relationship with allergic asthma in a population of Hans from Hubei province of China. METHODS: The polymorphisms (8570G > A and 11515C > A) were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 145 cases of allergic asthma and 130 healthy controls. The genotype and allele frequencies were calculated and analyzed. RESULTS: The genotype frequencies of GG, GA and AA in 8570G > A polymorphism were 0.985, 0.015 and 0 respectively in the healthy population and 0.931, 0.069 and 0 respectively in the allergic asthma population. There was significant difference in genotype and allele frequencies between the allergic asthma patients and control subjects (P=0. 030, P=0.032). The polymorphism of 11515C>o A was not detected. CONCLUSION: The polymorphism of 8570G > A in TIM4 may be associated with allergic asthma in the population of Han nationality from Hubei province of China. PMID- 17407087 TI - [Meiotic segregation results of male reciprocal chromosome translocations]. AB - OBJECTIVE: To analyze the meiotic segregation results of male reciprocal chromosome translocation by fluorescence in situ hybridization (FISH). METHODS: Multi-color FISH using 3 combined probes located in any 3 chromosome segments on both sides of two breakpoints was performed on the de-condensed sperm head to analyze the sperm chromosomal contents and segregation patterns. RESULTS: Four male reciprocal translocation carriers were included in the study, with the karyotypes of 46, XY, t(2;18) (p16; q23); 46, XY, t(4;6) (q34;q21); 46, XY, t(8;13) (q23;q21) and 46, XY, t(4;5) (4q31;5q13), respectively. The results showed that 4 carriers had different proportions of various segregated spermatozoa. The spermatozoa of alternate, adjacent-1, adjacent-2, 3:1, non disjunction in meiosis II, and 4:0 or diploidy accounted for 27.1%-49.4%, 26.9% 37.6%, 2.7%-15.7%, 8.6%-32.7%, 0.2%-1.9%, and 0.1%-0.4%, respectively. CONCLUSION: For each-reciprocal translocation carrier seems to have a particular meiotic segregation results, FISH analysis on sperm head should be done for each carrier in order to provide an accurate genetic counseling. PMID- 17407088 TI - [ssociation of HLA class I and II alleles with generalized vitiligo in Chinese Hans in north China]. AB - OBJECTIVE: To investigate the association of HLA class I and II alleles with generalized vitiligo in ethnic Han Chinese in north China. METHODS: By employing polymerase chain reaction sequence-specific primer (PCR-SSP) procedure 34 generalized vitiligo patients in north China were studied for HLA I and II alleles and were compared with 102 healthy controls. RESULTS: The allelic frequencies of HLA-A*30, Cw*06, DRB1*07, and DQB1*0201 were increased significantly in generalized vitiligo and especially in the patients without family history compared with the controls. CONCLUSION: These alleles positively associated with generalized vitiligo in Chinese Han patients in north China, might provide clues to reveal the susceptibility gene(s) of vitiligo in Chinese and as well as the immunnogenetic mechanisms of disease. PMID- 17407089 TI - [Clinical characteristics and spastin gene mutation analysis on an autosomal dominant kindred with hereditary spastic paraplegia]. AB - OBJECTIVE: To investigate the clinical characteristics and analyze spastin gene mutation on a kindred with hereditary spastic paraplegia (HSP). METHODS: All family members were studied through clinical examinations. The proband and another two patients in this kindred were subjected to electromyography (EMG) examinations. The proband was subjected to thoracic MRI examination too. Mutation analysis of spastin gene was screened by polymerase chain reaction combined with DNA sequencing in the proband and his father. RESULTS: All patients in the kindred manifested as classical HSP. Thoracic MRI revealed atrophies of the spinal cord in the proband. No abnormal spastin gene mutation was detected in these two patients. CONCLUSION: This kindred has typical clinical manifestations of HSP. The pathogenesis has no association with mutation of the exons of spastin gene. PMID- 17407090 TI - [Familial and genetic study in a large Chinese kindred with hereditary nonpolyposis colorectal cancer]. AB - OBJECTIVE: Hereditary nonpolyposis colorectal cancer (HNPCC) is one of the most common hereditary colon cancer syndromes accounting for 1%-5% of all colorectal cancer cases. Germline mutations in at least five genes coding for DNA mismatch repair (MMR) proteins are associated with the clinical phenotype of HNPCC. More than 400 MMR mutations have been identified in HNPCC patients, and about 40% of mutations affect MSH2 gene including nucleotide substitutions, deletions, and insertions. Only a few mutations have been reported in Chinese families. METHODS: A Chinese family with HNPCC was collected and peripheral blood of individuals from the family was obtained. Mutation analysis was performed on genomic DNA. RESULTS: The family fulfilled Amsterdam criteria I, and 17 people out of 31 were diagnosed as malignant tumor for 21 times. Twelve people (70.6%) had rectal cancer, and the onset age was young with an average of 42.9 years old. Right side colon cancer was common in the family. A novel duplication mutation of four nucleotides in exon 7 MSH2 (MSH2: c.1215_1218dupCCGA) was found, which result in a premature stop 10 codons downstream in MSH2 (p.L407fsX417) in the family. Site specific PCR was applied to the pre-symptomatic diagnosis. CONCLUSION: This novel genomic mutation MSH2 was confirmed to be pathogenic, and polymerase chain reaction with modified primer was successfully applied to the pre-symptomatic diagnosis. These data expand the spectrum MSH2 mutations causing HNPCC. PMID- 17407091 TI - [Association of polymorphisms of 1772 (C-->T) and 1790 (G-->A) in HIF1A gene with hypoxia adaptation in high altitude in Sherpas]. AB - OBJECTIVE: To investigate the association of single nucleotide polymorphisms (SNPs) of 1772 (C-->T) and 1790 (G-->A) in exon 12 of hypoxia-inducible factor 1, alpha subunit gene (HIF1A) with hypoxia adaptation in high altitude in Sherpas. METHODS: The blood samples were chosen from 148 Sherpas in Tibet high altitude and 90 Han nationality healthy people in Guangdong province, and from which genomic DNA was extracted. The single nucleotide polymorphisms of 1772(C-->T) and 1790(G-->A) in exon 12 of HIF1A gene were examined by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). RESULTS: The genotype frequency of HIF1A gene 1790 (G-->A) in Sherpas and Han nationality was 57.43% versus 75.56% in GG genotype, 37.84% versus 21.11% in GA genotype and 4.73% versus 3.33% in AA genotype. GG genotype frequency in Sherpas was lower than that in Han nationality (P<0.01), while GA genotype frequency in Sherpas was higher than that in Han nationality (P<0.01). No significant difference in CC, CT and TT genotype frequency of 1772(C-->T) was shown between two groups respectively. The total frequency of CC + GA, CT + AA, TT + GA and TT + AA in Sherpas was higher than that in Han nationality. CONCLUSION: Polymorphisms of HIF1A gene 1790 (G- >A) are associated with hypoxia adaptation in high altitude in Sherpas. GA and AA genotype may be benefit to hypoxia adaptation, and it is worthy of deep-going investigation. PMID- 17407092 TI - Orthogonal natural atomic orbitals form an appropriate one-electron basis for expanding CASSCF wave functions into localized bonding schemes and their weights. AB - Localized bonding schemes and their weights have been obtained for the pi electron system of nitrone by expanding complete active space self-consistent field wave functions into a set of Slater determinants composed of orthogonal natural atomic orbitals (NAOs) of Weinhold and Landis (Valency and Bonding: A Natural Bond Orbital Donor-Acceptor Perspective, 2005). Thus, the derived bonding schemes are close to orthogonal valence bond structures. The calculated sequence of bonding scheme weights accords with the sequence of genuine resonance structure weights derived previously by Ohanessian and Hiberty (Chem Phys Lett 1987, 137, 437), who employed nonorthogonal atomic orbitals. This accord supports the notion that NAOs form an appropriate orthogonal one-electron basis for expanding complete active space self-consistent field wave functions into meaningful bonding schemes and their weights. PMID- 17407093 TI - Can a physics-based, all-atom potential find a protein's native structure among misfolded structures? I. Large scale AMBER benchmarking. AB - Recent work has shown that physics-based, all-atom energy functions (AMBER, CHARMM, OPLS-AA) and local minimization, when used in scoring, are able to discriminate among native and decoy structures. Yet, there have been only few instances reported of the successful use of physics based potentials in the actual refinement of protein models from a starting conformation to one that ends in structures, which are closer to the native state. An energy function that has a global minimum energy in the protein's native state and a good correlation between energy and native-likeness should be able to drive model structures closer to their native structure during a conformational search. Here, the possible reasons for the discrepancy between the scoring and refinement results for the case of AMBER potential are examined. When the conformational search via molecular dynamics is driven by the AMBER potential for a large set of 150 nonhomologous proteins and their associated decoys, often the native minimum does not appear to be the lowest free energy state. Ways of correcting the potential function in order to make it more suitable for protein model refinement are proposed. PMID- 17407094 TI - Joint neighbors approximation of macromolecular solvent accessible surface area. AB - A new method for approximate analytical calculations of solvent accessible surface area (SASA) for arbitrary molecules and their gradients with respect to their atomic coordinates was developed. This method is based on the recursive procedure of pairwise joining of neighboring atoms. Unlike other available methods of approximate SASA calculations, the method has no empirical parameters, and therefore can be used with comparable accuracy in calculations of SASA in folded and unfolded conformations of macromolecules of any chemical nature. As shown by tests with globular proteins in folded conformations, average errors in absolute atomic surface area is around 1 A2, while for unfolded protein conformations it varies from 1.65 to 1.87 A2. Computational times of the method are comparable with those by GETAREA, one of the fastest exact analytical methods available today. PMID- 17407095 TI - Dynamic modelling in a study of surgical error management. AB - We demonstrate the use of dynamic longitudinal models to investigate error management in cardiac surgery. Case study data were collected from a multicentre study of the neonatal arterial switch operation (ASO). Information on two types of negative events, or 'errors', observed during surgery, major and minor events, was extracted from case studies. Each event was judged to be recovered from (compensated) or not (uncompensated). The aim of the study was to model compensation given the occurrence of past events within a case. Two models were developed, one for the probability of compensating for a major event and a second model for the probability of compensating for a minor event. Analyses based on dynamic logistic regression models suggest that the total number of preceding minor events, irrespective of compensation status, is negatively related with the ability to compensate for major events. The alternative use of random effects models is investigated for comparison purposes. PMID- 17407096 TI - Interaction of HLA-B27 homodimers with KIR3DL1 and KIR3DL2, unlike HLA-B27 heterotrimers, is independent of the sequence of bound peptide. AB - HLA-B27 can form beta-2 microglobulin (beta2m)-associated heterotrimers (HLA-B27) and beta2m-free homodimers (B27(2)). Here, we study the role of complexed peptide in the interaction of these forms of B27 with the killer cell immunoglobulin (Ig) like receptors KIR3DL1 and KIR3DL2 and with Ig-like transcripts LILRB1 and LILRB2. HLA-B27 tetramers complexed with three of five different naturally processed self peptides and three of seven pathogen-derived epitopes bound to KIR3DL1-expressing transfectants and NK cells. Heterotrimeric complexes containing peptides with charged amino acids at position 8 did not bind to KIR3DL1; however, studies with analogue peptides demonstrated that these are not the only peptide residues involved in binding. KIR3DL1 ligation by HLA-B27 inhibited NK cell IFN-gamma production in a peptide-dependent fashion. B27 but not HLA-A2, B7 or B57 heavy chains formed homodimers in the presence of peptide epitopes. B27(2) bound to KIR3DL1, KIR3DL2 and LILRB2 but not LILRB1. KIR3DL2 ligation by B27(2) inhibited NK and T cell IFN-gamma production. By contrast with HLA heterotrimers, B27(2) binding to KIR did not depend on the sequence of the bound peptide. Differences in KIR binding to classical HLA and B27(2) could be involved in the pathogenesis of spondyloarthritis. PMID- 17407097 TI - A role for plasmacytoid dendritic cells in the rapid IL-18-dependent activation of NK cells following HSV-1 infection. AB - Natural killer (NK) cells play a crucial role in the initial response to viral infections but the mechanisms controlling their activation are unclear. We show a rapid and transient activation of NK cells that results in the production of IFN gamma immediately following infection with herpes simplex virus type 1 (HSV-1). Activation of NK cells leading to synthesis of IFN-gamma was not mediated by a direct interaction with virus but required the presence of additional cell types and was largely dependent on the cytokine IL-18, but not IL-12. HSV-1-induced IFN gamma expression by NK cells in vitro was impaired in spleen cultures depleted of CD11c(+) cells. Conversely, coculture of NK cells with virus-exposed conventional DC or plasmacytoid (p)DC restored the production of IFN-gamma, indicating that multiple DC subsets could mediate NK cell activation. While conventional DC populations stimulated NK cells independently of IL-18, they were less effective than pDC in promoting NK cell IFN-gamma expression. In contrast, the potent stimulation of NK cells by pDC was dependent on IL-18 as pDC from IL-18-deficient mice only activated a similar proportion of NK cells as conventional DC. These data identify IL-18 as a crucial factor for pDC-mediated NK cell regulation. PMID- 17407098 TI - NADPH oxidase of human dendritic cells: role in Candida albicans killing and regulation by interferons, dectin-1 and CD206. AB - Human monocyte-derived DC express the enzyme NADPH oxidase, responsible for ROS production. We show that Candida albicans did not activate NADPH oxidase in DC, and was poorly killed by these cells. However, Candida-killing activity increased upon DC stimulation with the NADPH oxidase activator PMA and was further enhanced by DC treatment with IFN-alpha or IFN-gamma. This fungicidal activity took place at high DC-to-Candida ratio, but decreased at low DC-to-yeast ratio, when Candida inhibited the NADPH oxidase by contrasting the assembly of the enzyme on DC plasma membrane. The NADPH oxidase inhibitor diphenyliodonium chloride abrogated the PMA-dependent DC candidacidal capacity. Engagement of beta-glucan receptor dectin-1 induced NADPH oxidase activation in DC that was depressed by mannose binding receptor CD206 co-stimulation. Candida was internalized by DC through mannose-binding receptors, but not through dectin-1, thus explaining why Candida did not elicit NADPH oxidase activity. Our results indicate that NADPH oxidase is involved in DC Candida-killing activity, which is increased by IFN. However, Candida escapes the oxidative damage by inhibiting NADPH oxidase and by entering DC through receptors not involved in NADPH oxidase activation. PMID- 17407099 TI - Mature DC from skin and skin-draining LN retain the ability to acquire and efficiently present targeted antigen. AB - Skin-draining LN contain several phenotypically distinguishable DC populations, which may be immature or mature. Mature DC are generally considered to have lost the capacity to acquire and present newly encountered Ag. Using antibody opsonized liposomes as Ag carriers, we show that mature DC purified from skin explants are able to efficiently capture liposomes, process Ag encapsulated within them and activate Ag-specific CD4(+) T cells. Explant DC from mice with Langerhans cells (LC) expressing the primate diphtheria toxin receptor that were exposed to diphtheria toxin in vivo presented Ag as well as explant DC from wild type mice, indicating that LC are not required and dermal DC are probably responsible for this presentation. We further show that all DC subtypes from LN that capture opsonized Ag are capable of cross-presenting it to CD8(+) T cells. Induction of additional maturation in vivo by LPS or treatment with double stranded RNA did not alter the Ag presentation capacity of the skin or LN DC subtypes. These results suggest that mature DC present in skin-draining LN may play an important role in the induction of primary and/or secondary immune responses against Ag delivered to the LN that they take up by receptor-mediated endocytosis. PMID- 17407100 TI - Hereditary complement C7 deficiency in nine families: subtotal C7 deficiency revisited. AB - Deficiencies in terminal complement components, including the component C7, are uncommon and associated with an increased risk of recurrent systemic neisserial infection. A total of 22 molecular defects have been reported in the C7 gene with both complete (C7Q0) and subtotal (C7SD) C7 deficiencies. In this study we report the molecular basis of nine new cases of C7 deficiencies that were characterized by exon-specific sequence analysis. Seven different C7 gene mutations were identified corresponding to small deletions (n=2), splice site changes (n=1) and single base pair substitutions leading to nonsense (n=1) or missense (n=3) mutations. Altogether, three changes of the C7 gene (G357R, R499S and 5' splice donor site of intron 16) account for half of the molecular defects which emphasize that a restricted number of molecular abnormalities are involved in this deficiency. We identified two patients with combined C7Q0/C7SD(R499S) and established the C7SD(R499S) frequency at about 1% in normal Caucasian population. We demonstrated that C7(R499S) mutant protein is retained in the endoplasmic reticulum whereas the wild-type C7 is located in the Golgi apparatus. Our results provide evidence that R499S represents a loss-of-function polymorphism of C7 due to a defective folding of the protein. PMID- 17407101 TI - Blockade of TREM-2 exacerbates experimental autoimmune encephalomyelitis. AB - Triggering receptor expressed on myeloid cells (TREM-2) is a membrane receptor associated with DAP12 that is expressed primarily in myeloid cells, including dendritic cells and microglia, and promotes fusion of osteoclast precursors into multinucleated cells. A rare autosomal recessive condition, Nasu-Hakola disease (NHD) is associated with loss-of-function mutations in DAP12 and TREM-2. The brain pathology observed in NHD patients suggests that disruption of the TREM 2/DAP12 pathway leads to neurodegeneration with demyelination and axonal loss. In this study, we have characterized TREM-2 protein expression on microglia using a newly produced monoclonal antibody directed against the mouse TREM-2 receptor. We report that TREM-2 expression is up-regulated in the spinal cord during both the early inflammatory and chronic phases of myelin oligodendrocyte glycoprotein (MOG)(35-55)peptide-induced experimental autoimmune encaphalomyelitis (EAE). We also demonstrate that TREM-2 is highly expressed on microglial cells in the central nervous system (CNS) during EAE and that blockade of TREM-2 during the effector phase of EAE results in disease exacerbation with more diffuse CNS inflammatory infiltrates and demyelination in the brain parenchyma. These results demonstrate a critical role for TREM-2 during inflammatory responses in the CNS. PMID- 17407102 TI - GITR/GITRL: more than an effector T cell co-stimulatory system. AB - Glucocorticoid-induced TNFR-related protein (GITR) is a member of the TNFR superfamily, expressed in several cells and tissues including T lymphocytes, NK cells and antigen-presenting cells (APC). GITR activation, upon interaction with its ligand (GITRL), functions as a co-activating signal. GITRL is mainly expressed on APC and GITR/GITRL interaction is important for the development of immune response. This review summarizes recent results about the GITR/GITRL system, focusing on the interplay between APC, effector and regulatory T cells. PMID- 17407103 TI - Hierarchical assembly of TiO2 nanoparticles on WS2 nanotubes achieved through multifunctional polymeric ligands. PMID- 17407104 TI - The role of patient personality in the identification of depression in older primary care patients. AB - BACKGROUND: Our aim was to evaluate whether personality factors significantly contribute to the identification of depression in older primary care patients, even after controlling for depressive symptoms. METHODS: We examined the association between personality factors and the identification of depression among 318 older adults who participated in the Spectrum study. RESULTS: High neuroticism (unadjusted Odds Ratio (OR) 2.36, 95% Confidence Interval (CI) [1.42, 3.93]) and low extraversion (adjusted OR 2.24, CI [1.26, 4.00]) were associated with physician identification of depression. Persons with high conscientiousness were less likely to be identified as depressed by the doctor (adjusted OR 0.45, CI [0.22, 0.91]). CONCLUSION: Personality factors influence the identification of depression among older persons in primary care over and above the relationship of depressive symptoms with physician identification. Knowledge of personality may influence the diagnosis and treatment of depression in primary care. PMID- 17407105 TI - Ensemble-docking approach on BACE-1: pharmacophore perception and guidelines for drug design. AB - Beta-secretase (BACE-1), a key enzyme in the etiopathogenesis and progression of Alzheimer disease, is the focus of medicinal chemistry efforts both in the pharmaceutical industry and in academia. Despite the availability of diverse peptidomimetic BACE-1 inhibitors, nonpeptidic compounds suitable for oral delivery and transport across the blood brain barrier are in great demand. Herein, a number of active and structurally diverse inhibitors were selected and subjected to an ensemble-docking process into five BACE-1 X-ray structures. The calculated bioactive conformations of these inhibitors allowed us to build an exhaustive pharmacophore model, which captures both the common geometric and electronic features essential for enzyme inhibition. The model is intended to aid the rational design of new BACE-1 inhibitors. Furthermore, a comparison of BACE/cathepsin D X-ray structures was made to provide guidelines for the design of BACE-selective inhibitors. PMID- 17407106 TI - The role of interstitial brachytherapy with salvage surgery for the management of recurrent head and neck cancers. AB - BACKGROUND: The role of interstitial brachytherapy (IBT) in the salvage surgical management of regional lymphatic metastases for head and neck cancer has not been defined to date. To further explore its therapeutic benefit in the management of head and neck squamous cell carcinoma, the authors reviewed their experience utilizing IBT at the M. D. Anderson Cancer Center. METHODS: A retrospective chart review of all patients who were received IBT for cervical recurrences of head and neck cancer was undertaken. Twenty-two patients were identified; all patients underwent neck dissection and intraoperative placement of afterloading catheters as part of the salvage treatment. Soft tissue reconstruction was performed when necessary. The duration and dosage of salvage brachytherapy was reviewed, and the overall disease-free survival rates were determined. RESULTS: All patients had been treated with external beam radiation therapy prior to recurrence, with an average dose of 65 grays (Gy), and 46% of patients had undergone prior neck dissection. The median time to regional recurrence after definitive treatment was 30 months. Soft tissue coverage of the surgical bed with a muscle flap, most commonly a pectoralis major flap, was performed in 19 of 22 patients. The median IBT dose was 60 Gy over a total duration of 4 days. Postoperative complications were few, and there were no perioperative deaths. Recurrences in the reirradiated necks occurred in 27% of patients. CONCLUSIONS: Regional recurrences are common after multimodality treatment for head and neck cancer and are challenging to manage. Although its role has not been defined clearly, salvage neck dissection with IBT is beneficial in the management of recurrent lymphatic metastases of upper aerodigestive tract carcinomas. PMID- 17407107 TI - Depression and chronic medical illnesses in Asian older adults: the role of subjective health and functional status. AB - OBJECTIVE: Depression in elderly is reportedly associated with a number of specific chronic illnesses. Whether each of these co-morbid associations results uniquely from disease-specific psychobiological responses or is mediated by non specific factors like subjective health and functional status is unclear. METHOD: Analysis of data of 2,611 community-dwelling Chinese aged 55 and older, including depressive symptoms defined by Geriatric Depression Scale score >or= 5 and self reports of specific chronic illnesses. RESULTS: The prevalence of depressive symptoms was 13.3%, lower in those without chronic illness (7.5%), and higher in those with illnesses (13.2-24.2%). Crude Odds Ratios (OR) were significantly elevated for hypertension, eye disorders, diabetes, arthritis, ischemic heart disease, asthma/COPD, stroke, osteoporosis, heart failure, thyroid problem, and gastric problem. In multivariable analyses, only asthma/COPD [OR:2.85, 95% Confidence Intervals (CI): 1.36, 5.98], gastric problem (OR:2.64, 95% CI: 1.11, 6.29), arthritis (OR:1.87, 95% CI: 1.02, 3.42) and heart failure (OR:2.11, 95% CI: 0.98, 4.58) remained independently associated with depressive symptoms, after adjusting for comorbidities, subjective health and functional status, cognitive functioning, smoking, alcohol, psychosocial and demographic variables. CONCLUSION: Most comorbid associations of depressive symptoms with specific chronic illnesses are explained by accompanying poor self-reported health and functional status, but some illnesses probably have a direct psychobiological basis. PMID- 17407108 TI - Oligospiroketals as novel molecular rods. AB - A modular approach for the synthesis of molecular rods based on oligospiroketals has been developed. The strategy relies on different terminal and intermediate segments, which are joined by ketal formation between ketones and diols. For this purpose it was necessary to develop a new ketalization method to circumvent some problems related with the established methods. The terminal segments are either derived from 4-piperidinone or from 4-oxocyclohexane carboxylic acid whereas the intermediate segments rest on pentaerythritol and cyclohexane-1,4-dione. A series of trispiro (14-18), hexaspiro (19) and nonaspiro (20) compounds have been prepared and characterized. From these we realized that it is imperative to use solubility enhancing groups if more than seven rings are joined. PMID- 17407109 TI - Relaxometric and modelling studies of the binding of a lipophilic Gd-AAZTA complex to fatted and defatted human serum albumin. AB - A new lipophilic gadolinium chelate consisting of a long aliphatic chain bound to the AAZTA coordination cage (Gd-AAZTAC17) has been synthesised. It possesses two coordinated water molecules (q=2) in fast exchange with the solvent (tau298(M) = 67 ns), which yields a relaxivity of 10.2 mM(-1) s(-1). At concentrations greater than 0.1 mM, it forms micelles (average diameter 5.5 nm) characterised by a relaxivity of approximately 30 mM(-1) s(-1) at 20 MHz and 298 K. The latter value appears to be "quenched" by magnetic interactions among the Gd(III) ions on the surface of the micelle that cause a decrease in the electronic relaxation time. A relaxivity of 41 mM(-1) s(-1) was recorded for this micellar system when 98 % of the Gd(III) ions were replaced by diamagnetic Y(III). Gd-AAZTAC17 exhibits a better affinity for fatted human serum albumin (HSA) than for defatted HSA, whereas the relaxivities of the supramolecular adducts are reversed. The relaxivity shown by Gd-AAZTAC17/defatted HSA ({r b(1) (20 MHz, 298 K)=84 mM(-1) s(-1)) is by far the highest relaxivity reported so far for non-covalent paramagnetic adducts with slow-moving substrates. As shown by molecular docking calculations, the gadolinium complex enters a hydrophobic pocket present in fatted HSA more extensively than the corresponding adduct with defatted HSA. Interestingly, no marked difference was observed in either the relaxation enhancement or the binding affinity between fatted and defatted HSA when the binding titrations were carried out at a Gd-AAZTAC17 concentration higher than its critical micellar concentration (cmc). This behaviour has been attributed to the formation of an association between the negatively charged micelle of the lipophilic metal complexes and the positive residues on the surface of the protein. PMID- 17407110 TI - A smart surface in a microfluidic chip for controlled protein separation. AB - The smart surface created in a microfluidic chip has shown the capability of adsorbing and releasing proteins under electrical control. The inner surface of the chip channel was first coated by a thin layer of Au through sputtering and was subsequently modified with loosely packed self-assembled monolayers (SAMs) of thiols with terminal carboxylic or amino groups. Upon application of an external electric potential to the gold substrate, reversible conformational transformation between "bent" and "straight" states for the anchored mercapto chains could be modulated, through the electrostatic effect between the ionized terminal groups and the charged gold substrate. Thus, a hydrophobic or hydrophilic channel surface was established and could be reversibly switched electrochemically. Accordingly, the microchips prepared in this way can reversibly and selectively adsorb and release differently charged proteins under electrical control. Two model proteins, avidin and streptavidin, were demonstrated to be readily adsorbed by the smart chips under negative and positive potential, respectively. Also, more than 90 % of the adsorbed proteins could be released upon an electrical command. Furthermore, these chips were applied to the controlled separation of avidin and streptavidin mixtures with 1:1 and 1:1000 molar ratios. Under specific applied potentials, the chips adsorbed a certain protein from the mixture whereas the other protein was allowed to flow out, after which the adsorbed protein could be released by switching the applied potential. Thus, two eluted protein fractions were obtained and the separation of the two proteins was achieved. For the former mixture, each eluted fraction contained up to approximately 80-90 % avidin or streptavidin. For the latter mixture, the resulting separation efficiency indicated that the molar ratio of avidin and streptavidin could be increased from 1:1000 to about 32:1 after five run separations. PMID- 17407111 TI - Highly interpenetrated supramolecular networks supported by N...I halogen bonding. AB - Halogen bonding (XB) has been used to assemble tetrakis(4-pyridyl)pentaerythritol (tetradentate XB acceptor) with different alpha,omega-diiodoperfluoroalkanes (bidentate XB donors) or tetrakis(4-iodotetrafluorophenyl)pentaerythritol (tetradentate XB donor). The remarkable linearity of the XB formed, the rodlike character of alpha,omega-diiodoperfluoroalkanes and the mutual complementarities of pentaerythritol partners, translate the three-dimensional character of the XB acceptor into open primary networks, which interpenetrate to avoid the presence of voids and to ensure segregation of the modules. Two-dimensional (2D) square 4(4) layers (sql) with fourfold and fivefold interpenetration, as well as an eightfold diamondoid network (dia) of class Ia and a remarkable tenfold dia network of class IIIa, have been obtained. PMID- 17407112 TI - A comparison of phosphaferrocene and phospharuthenocene ligands in Rh+-catalysed enamide hydrogenation reactions: superior performance of the phospharuthenocene. AB - Enantiopure Cp*-substituted 3,4-dimethyl-5-phenylphosphametallocene-2-methanols (M=Fe, Ru) have been prepared from the corresponding 2-carboxy-(-) menthylphospholide anion and elaborated into 2-CH(2)PPh(2) phosphametallocenes (13: M=Fe; 14: M=Ru) and 2-CH(2)PtBuR substituted phospharuthenocenes (R=tBu, Me). The crystal structures of complexes [Rh(1,5-cod)(eta(2)-L)](+)BF(4)(-) (L=13, 14) reveal significantly different aryl group configurations. Comparative studies of the hydrogenation of para-substituted N-acetylcinnamate esters with these pre-catalysts show a superior performance for the phospharuthenocene derivative in terms of both rate and enantioselectivity. PMID- 17407113 TI - Organogels of 8-quinolinol/metal(II)-chelate derivatives that show electron- and light-emitting properties. AB - 8-quinolinol/copper(II)-, palladium(II)-, and platinum(II)-chelate-based organogelators (1 M) and their nongelling reference compounds (2 M) were synthesized. Complexes 1 M could gelate various organic solvents at very low concentrations. Electron microscope measurements gave visual images of well developed fibrous structures characteristic of low-molecular-weight organogels. UV/Vis and FTIR spectroscopy revealed that the good gelation ability of 1 M arises from the pi-pi interactions of the chelate moieties and the hydrogen-bond interactions among the amide groups. Very interestingly, field emission performances of the nanofibers prepared from the 1 M gels are evidently different depending on the electronic states of the three kinds of central metals. In addition, the 1 Pt gel shows unique thermo- and solvatochromism of visible and phosphorescent color in response to a sol-gel phase transition. Furthermore, the 1 Pt gel possesses an attractive ability to inhibit dioxygen quenching of excited triplet states, which increases the phosphorescence quantum yield of this gel. This effect is attributed to the isolation effect of the phosphorescent chelate moiety from the dioxygen-containing solution phase. PMID- 17407114 TI - Microwave effect in the fast synthesis of microporous materials: which stage between nucleation and crystal growth is accelerated by microwave irradiation? AB - Microporous materials, such as silicalite-1 and VSB-5 molecular sieves, have been synthesized by both microwave irradiation (MW) and conventional electric heating (CE). The accelerated syntheses by microwave irradiation can be quantitatively investigated by various heating modes conducted in two steps such as MW-MW, MW CE, CE-MW, and CE-CE (in the order of nucleation-crystal growth). In the case of synthesis by MW-CE or CE-MW, the heating modes were changed for the second step just after the appearance of X-ray diffraction peaks in the first step. We have quantitatively demonstrated that the microwave irradiation accelerates not only the nucleation but also crystal growth. However, the contribution to decrease the synthesis time by microwave irradiation is larger in the nucleation stage than in the step of crystal growth. The crystal size increases in the order of MW-MW30 x 10(9)/L, constitutional symptoms, circulating blasts > or = 1%, platelet count <100 x 10(9)/L, absolute monocyte count > or = 1 x 10(9)/L) identified all but platelet count as being significant. The Mayo PSS, which is based on the 4 CBC-derived parameters (ie, hemoglobin, platelet, leukocyte, and monocyte counts), displayed a better hazard ratio profile compared with the other 2 PSSs in identifying long-lived patients as well as delineating intermediate-risk disease category. The latter effect was even more pronounced in patients younger than age 60 years. CONCLUSIONS: The Mayo PSS for survival in PMF is an objective CBC-derived prognostic model that might be superior in its performance over that of either the Dupriez and Cervantes PSSs. PMID- 17407135 TI - Proportion of long-term event-free survivors and lifetime of adult patients not cured after a standard acute lymphoblastic leukemia therapeutic program: adult acute lymphoblastic leukemia-94 trial. AB - BACKGROUND: In adult acute lymphoblastic leukemia, treatment results generally are expressed in terms of overall survival or disease-free survival at 3 years. In this investigation, the authors attempted to express the results in terms of the proportion of long-term disease-free survivors and in terms of lifetime in patients who developed recurrent disease or died. METHODS: Univariate and multivariate analyses were used to assess the influence of different covariates on the 2 result criteria in 922 participants in the Adult Acute Lymphoblastic Leukemia-94 multicenter trial. RESULTS: The proportion of long-term survivors was 21.5% (95% confidence interval [95% CI], 18.1-25.4%) and was higher in women than in men. The proportion decreased with increasing age, white blood cell count, and lactate dehydrogenase level. The lowest proportion was observed in patients ages 44 years to 55 years (11.4%; 95% CI, 7-17.9%) and in patients with the t(9;22) BCR-ABL karyotype (13.4%; 95% CI, 8.8-19.8%), and the highest proportion was observed in patients with the t(4;11) MLL-AF4 karyotype (31.3%; 95% CI, 18.2 48.3%). The mean expected lifetime of patients who were not cured was 11.4 months (95% CI, 9.1-14.1 months). It was longer in men than in women and was shorter with increasing age, performance status, hemoglobin level, and white blood cell count. CONCLUSIONS: The results of this study highlighted and specified the importance of some classic prognostic factors in patients with acute lymphoblastic leukemia. PMID- 17407136 TI - Most human Alu and murine B1 repeats are unique. AB - Alus and B1s are short interspersed repeat elements (SINEs) indirectly derived from the 7SL RNA gene. While most researchers recognize that there exists extensive variability between individual elements, the extent of this variability has never been systematically tested. We examined all Alu elements over 200 nucleotides and all B1 elements over 100 nucleotides in the human and mouse genomes, and analyzed the number of copies of each element at various stringencies from 22 nucleotides to full length. Over 98% of 923,277 Alus and 365,377 B1s examined were unique when queried at full length. When the criterion was reduced to half the length of the repeat, 97% of the Alus and 73% of the B1s were still found to be a single copy. All single and multi-copy sequences have been mapped and documented. Access to the data is possible using the AluPlus website http://www.ibr.hawaii.edu. PMID- 17407137 TI - Intracystic bleomycin therapy for craniopharyngioma in children: the Canadian experience. AB - BACKGROUND: Surgical removal and radiation therapy are associated with significant risk for morbidity in the pediatric population with craniopharyngioma. Intracystic therapies have been utilized in some centers to potentially decrease morbidity associated with cystic craniopharyngioma. The aim of the study was to review the Canadian experience with intracystic bleomycin therapy (ICB). METHODS: All centers in the Canadian Pediatric Brain Tumor Consortium (CPBTC) were invited to participate in a retrospective review of this treatment. A questionnaire was sent to each center. The data were analyzed at British Columbia's Children's Hospital. RESULTS: In all, 6 of 17 centers utilized ICB and submitted data. A total of 17 of 19 patients with the intention to treat received bleomycin. Twelve were treated at the time of diagnosis, and 5 at the time of recurrence. Five patients achieved a complete response, 6 achieved a partial response, and 5 achieved a minor response to bleomycin. One patient was stable for 2.8 years. At the time of last follow-up, 8 patients have not required further intervention. Complications included transient symptomatic peritumoral edema (2 patients), precocious puberty (1 patient) and panhypopituitarism (2 patients). The median follow-up was 4 years (range, 0.5-10.2 years). The median progression-free survival was 1.8 years (range, 0.3-6.1 years). One patient died of a massive infarct secondary to radiation-induced moyamoya syndrome. CONCLUSIONS: ICB was found to be well tolerated in this group of children. ICB may be a feasible and effective therapy for certain children with craniopharyngioma. Bleomycin may delay the need for aggressive surgery or radiation therapy for several years. Prospective multiinstitutional clinical trials are required to further evaluate the feasibility, effectiveness, and dose schedules of this treatment. PMID- 17407138 TI - Risk factors for malignancy in Japanese renal transplant recipients. AB - BACKGROUND: Among recipients of renal transplants, the incidences of renal cancer and gastrointestinal cancer are higher and that of skin cancer is much lower in Japan than in Europe and North America. METHODS: The risk factors for the development of malignant tumors were examined in Japanese recipients of renal transplants. A total of 556 patients underwent renal transplantation at the Department of Urology, Osaka University Faculty of Medicine between March 1, 1965, and April 31, 2004. Of these patients, 366 were retrospectively studied in whom risk factors potentially related to the development of malignancy could be evaluated on the basis of medical records. The incidence of malignancy, survival rate, and risk factors for malignancy were examined. RESULTS: The overall incidence of malignancy was 6.8% (25/366 patients). Six of the 25 patients with malignancy died of cancer, but there was no correlation between the occurrence of malignancy and the survival rate (P = .8058, log-rank test). A Cox proportional hazards model identified treatment with tacrolimus (hazard ratio [HR] = 4.376; 95% confidence interval [CI]: 1.647-11.627; P = .0031) and age at transplantation (HR = 1.562; 95% CI: 1.089-2.240; P = .0155) as risk factors for malignancy. CONCLUSIONS: The results of multivariate analysis suggested that age at transplantation and the use of tacrolimus were independent risk factors for the development of malignancy in recipients of renal transplants. PMID- 17407139 TI - A role for Tbx2 in the regulation of the alpha2(1) collagen gene in human fibroblasts. AB - The T-box gene family encodes highly conserved transcription factors that play important roles in embryonic development and have been implicated in carcinogenesis. One member of the family, Tbx2, is generally regarded as a transcriptional repressor but appears to be capable of functioning as an activator depending on the cellular context. This study shows that Tbx2 is expressed in normal human fibroblasts but is drastically reduced in several transformed fibroblast cell lines. This pattern of Tbx2 expression correlated with that observed for the human alpha2(1) collagen gene (COL1A2). Interestingly, stable expression of transfected Tbx2 in transformed fibroblast cell lines further reduces expression of the human endogenous COL1A2 gene. This ability of Tbx2 to repress the human COL1A2 gene was confirmed in luciferase reporter assays and shown to be independent of the consensus T-box binding element. PMID- 17407140 TI - Overexpression of integrin beta1 inhibits proliferation of hepatocellular carcinoma cell SMMC-7721 through preventing Skp2-dependent degradation of p27 via PI3K pathway. AB - Integrins may play important roles in many cellular events, such as cell proliferation, differentiation, and apoptosis. We showed previously that overexpression of integrin beta1 inhibits cell proliferation in SMMC-7721 cells. Here we reported that one of the cyclin-dependent kinase (CDK) inhibitors, p27(Kip1) was involved in proliferation-inhibition induced by overexpression of integrin beta1. Overexpression of integrin beta1 upregulated p27(Kip1) at the protein level, but not mRNA level. The knock-down of p27(Kip1) expression restored cell growth in integrin beta1-overexpressing cells. Cycloheximide (Chx) treatment and pulse-chase experiments revealed that overexpression of integrin beta1 prolonged the half-life of p27(Kip1) by inhibiting its degradation. Proteasome inhibitor (MG132) treatment of the cells indicated that proteasome mediated degradation of p27, and Skp2-dependent degradation might be prevented. Overexpression of integrin beta1 decreased Skp2 at mRNA level, which was regulated by cell adhesion and the subsequent adhesion-dependent signaling. Overexpression of integrin beta1 reduced cell adhesion, accordingly, inactivated the phosphoinositide 3-kinase (PI3K) signaling. PI3K inhibitor LY294002 upregulated p27(Kip1) at post-translational level and downregulate Skp2 at mRNA level, which could mimic the effects of integrin beta1 overexpression on p27(Kip1) and Skp2. Together, these results suggested that overexpression of integrin beta1 inhibited cell proliferation by preventing the Skp2-dependent degradation of p27(Kip1) via PI3K pathway. PMID- 17407141 TI - Intracoronary delivery of autologous bone marrow mononuclear cells radiolabeled by 18F-fluoro-deoxy-glucose: tissue distribution and impact on post-infarct swine hearts. AB - Intracoronary injection of the bone marrow-derived mononuclear cells (MNCs) is emerging as a potentially novel therapy for ischemic heart failure. This study was aimed at assessing the efficacy of intracoronary MNC delivery in the myocardium. The in vivo distribution and myocardial homing of intracoronarily delivered MNCs in experimental Chinese swine with acute myocardial infarction (AMI) created by occlusion of left anterior descending (LAD) coronary artery for 90 min. MNCs radiolabeled with 18F-fluoro-deoxy-glucose (18F-FDG) were delivered using a coronary catheter into the infarct-related coronary artery 1 week after AMI. Dual-nuclide single photon emission computed tomography (SPECT) revealed that 1 h after cell infusion, 6.8 +/- 1.8% of 18F-FDG-labeled MNCs occurred in the infarcted myocardium with the remaining activity found primarily in the liver and spleen. In the heart, MNCs were detected predominantly in the under-perfused myocardium. The infused cells retained in the hearts at a rate highly correlated with the under-perfused lesional sizes. Pathological examination further demonstrated that 6 weeks after infusion, compared to controls, the hearts receiving MNCs exhibited less fibrosis and inflammatory infiltrate, more viable tissue, and higher vascular density. Cardiac function was significantly improved in the MNC-infused hearts. Thus, 18F-FDG labeling and dual-nuclide SPECT imaging is capable of monitoring in vivo distribution and homing of MNCs after intracoronary infusion. MNC coronary delivery may improve cardiac function and positive ventricular remodeling in the heart with AMI. PMID- 17407142 TI - Mesothelioma in blood related subjects: report of 11 clusters among 1954 Italy cases and review of the literature. AB - BACKGROUND: Malignant mesothelioma is a sporadic tumor related to asbestos. Its occurrence in blood relatives raises the question of potential contribution of predisposing factors. METHODS: The study analyses the features of mesothelioma in blood relatives that might explain the disease clustering. Data sources of familial clusters were three population-based Mesothelioma Registries in Italy (Veneto and Apulia Regions, Brescia province; 1978-2005) and Medline, Toxline, and Oshline/Hseline databases for a review of the literature (1968-2006). RESULTS: Eleven clusters (22 cases) were identified among 1954 Italy mesothelioma cases, and 51 clusters (120 cases) were extracted from 33 studies. The proportion of Italy familial cases was 1.4 per 100 mesothelioma cases; the ratio between the number of familial clusters and the number of non-familial mesothelioma cases was 1:148. The mesothelioma profile in consanguineous is the same as in non consanguineous subjects (male prevalence; pleural site; age at diagnosis >50 years; asbestos exposure). Most clusters occurred in asbestos workers (shipyard, asbestos-cement production/processing, and insulation) and household-exposed blood relatives. Others were related to asbestos-cement factory pollution, asbestos-in-place, and handling asbestos-contaminated textiles. Two clusters were without any known exposure. Cancer family history revealed lung cancer cases in eight clusters. CONCLUSIONS: Available data support asbestos exposure as the main risk factor in mesothelioma cases among blood relatives. Our finding of a low proportion of familial cases would not suggest the influence of a large genetic component for mesothelioma in blood relatives. PMID- 17407143 TI - A brief biography of Bruce Merrifield: his life and legacy. PMID- 17407144 TI - Ascochlorin suppresses oxLDL-induced MMP-9 expression by inhibiting the MEK/ERK signaling pathway in human THP-1 macrophages. AB - The critical initiating event in atherogenesis involves the invasion of monocytes through the endothelial walls of arteries and the transformation of monocytes from macrophages into foam cells. Human THP-1 monocytic cells can be induced to differentiate into macrophages by phorbol myristate acetate (PMA) and can then be converted into foam cells by exposure to oxidized low-density lipoprotein (oxLDL). Also, during a chronic inflammatory response, monocytes/macrophages produce the 92-kDa matrix metalloproteinase-9 (MMP-9) that may contribute to the extravasation, migration, and tissue remolding capacities of the phagocytic cells. Here, we investigate the effect of ascochlorin (ASC), a prenylphenol antiviral compound from the fungus Ascochyta viciae, on oxLDL-induced MMP-9 expression and activity in human THP-1 macrophages. ASC reduced oxLDL-induced MMP 9 expression and activity in a time-dependent and dose-dependent manner. Also, an analysis of MMP-9 activity using pharmacologic inhibitors showed that ASC inhibits MMP-9 activity via the extracellular signal-regulated kinase 1 and kinase 2 pathways. Our results suggest that ASC may be useful as a potent clinical antiatherogenic agent, a topic of considerable interest in the biological chemistry of chemotherapeutic agents. PMID- 17407145 TI - Pesticide illness among flight attendants due to aircraft disinsection. AB - BACKGROUND: Aircraft "disinsection" is the application of pesticides inside an aircraft to kill insects that may be on board. Over a 1-year period, California's tracking system received 17 reports of illness involving flight attendants exposed to pesticides following disinsection. METHODS: Interviews, work process observations, and a records review were conducted. Illness reports were evaluated according to the case definition established by the National Institute for Occupational Safety and Health. RESULTS: Twelve cases met the definition for work related pesticide illness. Eleven cases were attributed to the "Residual" method of disinsection, i.e., application of a solution of permethrin (2.2% w/w), solvents (0.8%), and a surfactant (1.4%); the method of disinsection could not be determined for one case. CONCLUSIONS: The aerosol application of a pesticide in the confined space of an aircraft cabin poses a hazard to flight attendants. Nontoxic alternative methods, such as air curtains, should be used to minimize disease vector importation via aircraft cabins. Employers should mitigate flight attendant pesticide exposure in the interim. PMID- 17407146 TI - Nested case-control study of occupational chemical exposures and prostate cancer in aerospace and radiation workers. AB - BACKGROUND: To date, little is known about the potential contributions of occupational exposure to chemicals to the etiology of prostate cancer. Previous studies examining associations suffered from limitations including the reliance on mortality data and inadequate exposure assessment. METHODS: We conducted a nested case-control study of 362 cases and 1,805 matched controls to examine the association between occupational chemical exposures and prostate cancer incidence. Workers were employed between 1950 and 1992 at a nuclear energy and rocket engine-testing facility in Southern California. We obtained cancer incidence data from the California Cancer Registry and seven other state cancer registries. Data from company records were used to construct a job exposure matrix (JEM) for occupational exposures to hydrazine, trichloroethylene (TCE), polycyclic aromatic hydrocarbons (PAHs), benzene and mineral oil. Associations between chemical exposures and prostate cancer incidence were assessed in conditional logistic regression models. RESULTS: With adjustment for occupational confounders, including socioeconomic status, occupational physical activity, and exposure to the other chemicals evaluated, the odds ratio for low/moderate TCE exposure was 1.3; 95%CI = 0.8 to 2.1, and for high TCE exposure was 2.1; 95%CI = 1.2 to 3.9. Furthermore, we noted a positive trend between increasing levels of TCE exposure and prostate cancer (P-value for trend = 0.02). CONCLUSION: Our results suggest that high levels of TCE exposure are associated with prostate cancer among workers in our study population. PMID- 17407147 TI - Floodwater exposure and the related health symptoms among firefighters in New Orleans, Louisiana 2005. AB - BACKGROUND: Concerns over increased reports of physical health symptoms thought to be related to floodwater exposure among New Orleans firefighters prompted a health hazard evaluation of firefighters following Hurricane Katrina. METHODS: A questionnaire assessing health symptoms possibly related to the response to Hurricane Katrina was administered to all New Orleans Fire Department (NOFD) personnel within 3 months of the disaster. Descriptive statistics were compiled and prevalence ratios (PR) were estimated for covariates using generalized linear models with Log link and Poisson distribution. RESULTS: Of the 525 firefighters who completed the questionnaire (77% participation), 201 (38%) reported one or more new-onset respiratory symptoms, such as sinus congestion (145 [28%]), throat irritation (92 [17%]), and cough (124 [24%]). Skin rash was reported by 258 (49%) of respondents, 414 (79%) reported skin contact with floodwater, and 165 (32%) reported contact with floodwater on multiple days. In multivariate analyses adjusting for age, gender, and smoking, firefighters who had floodwater contact with skin and either nose/mouth or eyes (224, 44%) had an increased rate of new onset upper respiratory symptoms (PR = 1.9; 95% confidence interval [CI], 1.1, 3.1), and skin rash (PR = 2.1; 95% CI, 1.4, 3.2) compared to those not exposed to the floodwater. CONCLUSIONS: Response workers involved with floodwater should minimize direct skin and mucosal contact with floodwater if possible through the use of appropriate personal protective equipment, such as goggles, safety glasses with side shields, or full-face shields. PMID- 17407148 TI - Musculoskeletal symptoms among poultry processing workers and a community comparison group: Black women in low-wage jobs in the rural South. AB - BACKGROUND: Poultry processing is characterized by rapid line speed and extreme division of labor. Morbidity associated with this work has been reported by scientists, journalists and workers in this fast growing industry. METHODS: Cross sectional data from baseline measures of a cohort of black women employed in poultry processing (n = 291) and a community comparison group (n = 299) in rural North Carolina were used to evaluate musculoskeletal symptom reports and to explore factors associated with those reports. Recruitment of participants and collection of data were performed by women in the community circumventing the need to involve employers. RESULTS: Significant differences in musculoskeletal symptom prevalence were observed between women employed in poultry processing and those of similar economic status employed elsewhere in the same area of rural northeastern North Carolina. After adjusting for other factors independently associated with symptoms among these women including age, depression, and perceived isometric load at work, the prevalence of upper extremity and neck symptoms was 2.4 (95% CI 1.7, 3.2) times higher among women working in poultry processing. CONCLUSIONS: The findings add to the documentation of occupational health concerns among vulnerable workers employed in poultry processing in our state; in this economically depressed area of rural northeastern North Carolina poultry processing is the largest single employer of women. On a larger scale, the potential magnitude of upper extremity morbidity among women employed in poultry processing should be viewed with the knowledge that poultry processing is a growing industry in the U.S. with work done largely by blacks and immigrants. PMID- 17407149 TI - Hand preferences on unimanual and bimanual tasks in white-faced capuchins (Cebus capucinus). AB - This study examined hand preference in white-faced capuchins on a unimanual task and on a coordinated bimanual task. For the unimanual task, handedness was assessed by observing simple reaching for small grains. For the bimanual task, tubes lined with chocolate paste inside were presented to the capuchins. The hand and the finger(s) used to remove chocolate paste were recorded. Seven individuals out of eight in the reaching task and 12 out of 13 in the tube task exhibited a hand preference. Moreover, test-retest correlations showed stability in hand use across time for the coordinated bimanual task. We found no significant differences in strength of hand preference between sexes. Finally, as noted in other primate species, the capuchins were more lateralized in the bimanual task compared to the unimanual task. PMID- 17407150 TI - Age-dependent telomere-shortening is repressed by phosphorylated alpha-tocopherol together with cellular longevity and intracellular oxidative-stress reduction in human brain microvascular endotheliocytes. AB - Cellular life-span of neonatal human brain microvascular endotheliocytes (HBME) was estimated by population doubling levels (PDLs) for serial subcultivations until spontaneous proliferation stoppage, and was 2.4-fold longer for continuous administration with the 6-O-phosphorylated derivative (TocP) of alpha-tocopherol (Toc), being bio-available owing to its water-solubility, or TocP plus 2-O phosphorylated ascorbate (Asc2P), and 1.3-fold longer with Asc2P, at a dose of 150 microM, than for the non-administered control. Enlarged cell diameters indicative of cellular aging were repressed for TocP-administered cells as analyzed with a channelizer. Age-dependent shortening of telomeric DNA length (291 bp/PDL) was slowed markedly for TocP (165 bp/PDL) or TocP plus Asc2P, but slightly for Asc2P. Telomerase activity as assessed by the PCR-based TRAP method was detectable slightly at younger ages but no longer at middle ages for the non administered cells, but, for TocP-administered cells, was intensely detected at younger ages and appreciably until middle ages. Intracellular TocP amounts were not changed age-dependently in contrast to a marked decrease in Toc which accrued from TocP esterolysis. This may be partly attributed to age-dependent changes in the lipid peroxidation product acrolein (ACR), which was abundant at older ages in non-administered cells, but scarcely in TocP-administered cells. Furthermore, intracellular reactive oxygen species (ROS) such as H(2)O(2) and hydroperoxides as detected using the redox indicator CDCFH-DA was less abundant in TocP administered cells than in non-administered cells. Thus the telomeric-DNA retention, concurrently with retained telomerase activity, was shown to be correlated with cellular longevity, and may be supported by diminished oxidative stress, in hydrophobic microenvironment, which can be achieved by TocP rather than AscP. PMID- 17407151 TI - Ergonomics and regulatory politics: the Washington State case. AB - BACKGROUND: Every year in the State of Washington more than 50,000 workers experience a work related musculoskeletal disorder (WMSD), making up more than 30% of all worker compensation cases. In 2000, the Washington State Department of Labor and Industries (L&I) adopted a workplace ergonomics rule requiring employers to reduce worker exposure to hazards that cause or contribute to WMSDs. In 2003, the ergonomics rule was repealed by a margin of 53.5-46.5 in a statewide voter initiative. METHODS: The official rulemaking record of approximately 100,000 pages, along with supplementary published and unpublished material, was reviewed. The relationship between scientific deliberation and the public policy process in adopting and repealing the ergonomics rule was assessed and described. The deliberative features of the regulatory, judicial, legislative, and ballot processes were compared. RESULTS AND CONCLUSIONS: The ergonomics rule was successful in the regulatory and legal arenas where the process was most transparent and open to public involvement, differing views could be presented fully, and decision makers were expected to explain their decisions in light of the record. The rule fared most poorly in the legislature and at the ballot box when these features were lost and where considered deliberation was replaced by unconstrained political conflict. Additional checks and balances are needed. PMID- 17407152 TI - Studies on the cell treatment conditions to elicit lipolytic responses from 3T3 L1 adipocytes to TCDD, 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - Wasting syndrome is one of the hallmark symptoms of poisoning by TCDD (=dioxin), which is associated with the massive loss of adipose tissue and serum hyperlipidemia in vivo. Yet, the most widely used in vitro cell model 3T3-L1 adipocyte has not been useful for studying such an action of TCDD because of the difficulty of inducing their mature adipocytes to respond to TCDD to go through lipolysis. Here, we made efforts to find the right cell culture and treatment conditions to induce mature 3T3-L1 adipocytes to go through lipolysis, which is defined as events leading to reduction of lipids in adipocytes. The optimum condition was found to require 7-day differentiated adipocytes being subjected to DMEM medium containing TCDD (but without insulin) for 5 day incubation with two medium changes (the same composition) on incubation days 2 and 4. After 24 h, the early effect of TCDD on adipocytes was predominantly on inflammation, particularly induction of COX-2 and KC (IL-8), which is accompanied by upregulation of C/EBPbeta and delta. The sign of TCDD-induced lipolysis starts slowly and by incubation day 3, a few markers showed modestly significant changes. By day 5 of incubation, however, many markers show highly significant signs of lipolytic changes. Although this process could take place without exogenous macrophages or their cytokines, addition of exogenous TNFalpha considerably synergized this action of TCDD. In conclusion, under a right condition, 3T3-L1 adipocytes were found to respond to TCDD to go through lipolysis. The early trigger of such a response appears to be activation of COX 2, which is amplified by TNFalpha. PMID- 17407153 TI - Expression of cyclin D3 through Sp1 sites by histone deacetylase inhibitors is mediated with protein kinase C-delta (PKC-delta) signal pathway. AB - The histone deacetylase (HDAC) inhibitors are an exciting new class of drugs that are targeted as anti-cancer agents. These compounds can induce growth arrest, apoptosis, and/or terminal differentiation in a variety of cancers. The inhibition of HDACs shifts toward hyper-acetylation, thereby driving transcriptional activation. In present study, HDAC inhibitor apicidin was used to elucidate the effect on expression of cell cycle related proteins and the molecular mechanism for transcriptional regulation of cyclin D3 in response to HDAC inhibitors in human colon cancer cells. We found that apicidin increases the transcriptional activity of cyclin D3 gene, which results in accumulation of cyclin D3 mRNA and protein. Apicidin-induced cyclin D3 expression is mediated by Sp1 sites within the cyclin D3 promoter. Apicidin-mediated cyclin D3 expression is attenuated by rottlerin, a specific protein kinase C-delta (PKC-delta) inhibitor, but not mitogen-activated protein kinases (MAPKs) inhibitors. Furthermore, suppression of PKC-delta expression by transfection with its siRNA prominently attenuated apicidin-induced cyclin D3 expression. These results indicate that the cyclin D3 induction caused by apicidin was associated with PKC delta signaling pathway not MAPKs signaling pathways. Taken together, these results suggest that the activation of cyclin D3 transcription by HDAC inhibitor apicidin was mediated through Sp1 sites and pointed to the possible participation of PKC-delta. PMID- 17407154 TI - The transferrin receptor and the tetraspanin web molecules CD9, CD81, and CD9P-1 are differentially sorted into exosomes after TPA treatment of K562 cells. AB - Here we show that treatment of K562 cells with the phorbol ester TPA induces the down-modulation of various surface antigens. Among them, the transferrin receptor (TfR), the tetraspanin CD81, and a CD81-associated protein, CD9P-1, were unique in that their expression levels were lower after 24 h incubation than after 3 h. We demonstrated that like the TfR, CD81 was internalized at early times, and was less synthesized at latter times. Despite the association of a fraction of the TfR with CD81, these two molecules were subjected to different fates. TPA increased targeting of CD81 and CD9P-1 into exosomes but strongly reduced the localization of the TfR in these vesicles. Using this model we have shown that a fraction of CD81 and CD9P-1 in exosomes comes from a surface pool and that these molecules remain associated in exosomes. However, CD9P-1 could be targeted to exosomes in the absence of CD81 and of another tetraspanin, CD9. The targeting of CD9 into exosomes did not require palmitoylation of the protein. J. Cell. Biochem. 102: 650-664, 2007. (c) 2007 Wiley-Liss, Inc. PMID- 17407155 TI - Syndrome-causing mutations of the BLM gene in persons in the Bloom's Syndrome Registry. AB - Bloom syndrome (BS) is caused by homozygous or compound heterozygous mutations in the RecQ DNA helicase gene BLM. Since the molecular isolation of BLM, characterization of BS-causing mutations has been carried out systematically using samples stored in the Bloom's Syndrome Registry. In a survey of 134 persons with BS from the Registry, 64 different mutations were identified in 125 of them, 54 that cause premature protein-translation termination and 10 missense mutations. In 102 of the 125 persons in whom at least one BLM mutation was identified, the mutation was recurrent, that is, it was shared by two or more persons with BS; 19 of the 64 different mutations were recurrent. Ethnic affiliations of the persons who carry recurrent mutations indicate that the majority of such persons inherit their BLM mutation identical-by-descent from a recent common ancestor, a founder. The presence of widespread founder mutations in persons with BS points to population genetic processes that repeatedly and pervasively generate mutations that recur in unrelated persons. PMID- 17407156 TI - Tumor microenvironment. PMID- 17407157 TI - Antiresorptive agents and osteoclast apoptosis. AB - Antiresorptive agents have proven to be effective therapies for the treatment of bone diseases associated with excessive osteoclast activity. Decreased osteoclast formation, inhibition of osteoclast actions, and reduced osteoclast survival represent mechanisms by which antiresorptive agents could act. The goals of this article are to present the evidence that antiresorptive agents can decrease osteoclast survival through apoptosis, to review the mechanisms by which they are thought to activate the apoptotic process, and to consider whether the actions on apoptosis fully account for the antiresorptive effects. As background, the apoptotic process will be briefly summarized together with the evidence that factors that promote osteoclast survival affect steps in the process. Following this, therapeutic agents that are both antiresorptive and can stimulate osteoclast apoptosis will be discussed. Other bone therapeutic agents that are either antiresorptive or apoptotic, but not both, will be described. Finally, newer antiresorptive compounds that elicit apoptosis and could represent potential therapeutic agents will be noted. PMID- 17407158 TI - Regulation of matrix Gla protein by parathyroid hormone in MC3T3-E1 osteoblast like cells involves protein kinase A and extracellular signal-regulated kinase pathways. AB - Inhibition of osteoblast-mediated mineralization is one of the major catabolic effects of parathyroid hormone (PTH) on bone. Previously, we showed that PTH induces matrix gamma-carboxyglutamic acid (Gla) protein (MGP) expression and established that this induction is critical for PTH-mediated inhibition of osteoblast mineralization. In the present study, we focus on the mechanism through which PTH regulates MGP expression in osteoblastic MC3T3-E1 cells. Following transient transfection of these cells with a -748 bp murine MGP promoter-luciferase construct (pMGP-luc), PTH (10 (-7) M) induced promoter activity in a time-dependent manner with a maximal four- to six fold induction seen 6 h after PTH treatment. Both H-89 (PKA inhibitor) and U0126 (MEK inhibitor), suppressed PTH induction of MGP promoter activity as well as the MGP mRNA level. In addition, forskolin (PKA activator) stimulated MGP promoter activity and mRNA levels confirming that PKA is one of the signaling molecules required for regulation of MGP by PTH. Co-transfection of MC3T3-E1 cells with pMGP-luc and MEK(SP), a plasmid encoding the constitutively active form of MEK, led to a dose-dependent increase in MGP promoter activity. Both MGP promoter activity and MGP mRNA level were not affected by the protein kinase C (PKC) inhibitor, GF109203X. However, phorbol 12-myristate 13-acetate (PMA), a selective PKC activator induced MGP mRNA expression through activation of extracellular signal-regulated kinase (ERK). Taken together, these results indicate that PTH regulates MGP via both PKA- and ERK-dependent pathways. PMID- 17407159 TI - Elevated plasma tissue inhibitor of metalloproteinase-1 level predicts decreased response and survival in metastatic breast cancer. AB - BACKGROUND: Tissue inhibitors of metalloproteinase (TIMPs) have at least 2 different functions. They inhibit the catalytic activity of matrix metalloproteinases, and they act as growth factors. METHODS: Pretreatment ethylenediamine tetracetic acid plasma TIMP-1 was assayed from 251 patients who were enrolled in a Phase III, second-line, hormone therapy trial, and from a control group of 50 healthy, postmenopausal women by using the TIMP-1 enzyme linked immunosorbent assay. RESULTS: The plasma TIMP-1 levels from the postmenopausal control group (n = 50 women) were 201 +/- 86 ng/mL mean +/- standard deviation (range, 49-455 ng/mL). The upper limit of normal was defined as the mean +/- 2 standard deviations of the control group (373 ng/mL). Patient pretreatment plasma TIMP-1 levels ranged from 70 ng/mL to 982 ng/mL. Plasma TIMP 1 was elevated above the mean + 2 standard deviations of the control group (373 ng/mL) in 19 patients (7.6%). In univariate analysis, patients who had elevated versus normal plasma TIMP-1 levels had a reduced clinical benefit rate (CBR) (16% vs 42%; P = .03). The time to progression (TTP) (84 days vs 174 days; P < .0001) and overall survival (141 days vs 860 days; P = .0001) also were significantly shorter in patients who had elevated TIMP-1 levels. TTP and overall survival also were significantly shorter in patients who had higher TIMP-1 plasma levels when it was analyzed as a continuous variable. In multivariate analysis, elevated plasma TIMP-1 level remained a prognostic factor for reduced overall survival (P < .0001) along with elevated serum HER-2/neu (P < .0001) and the presence of visceral metastases (P = .008). CONCLUSIONS: Elevated pretreatment plasma levels of TIMP-1 predicted a decreased response to second-line hormone therapy and reduced survival in women with metastatic breast cancer. PMID- 17407160 TI - Principles of nanostructure design with protein building blocks. AB - Currently there is increasing interest in nanostructures and their design. Nanostructure design involves the ability to predictably manipulate the properties of the self-assembly of autonomous units. Autonomous units have preferred conformational states. The units can be synthetic material science based or derived from functional biological macromolecules. Autonomous biological building blocks with available structures provide an extremely rich and useful resource for design. For proteins, the structural databases contain large libraries of protein molecules and their building blocks with a range of shapes, surfaces, and chemical properties. The introduction of engineered synthetic residues or short peptides into these can expand the available chemical space and enhance the desired properties. Here we focus on the principles of nanostructure design with protein building blocks. PMID- 17407161 TI - Structural and functional characterization of binding sites in metallocarboxypeptidases based on Optimal Docking Area analysis. AB - The metallocarboxypeptidases (MCPs) belonging to the clan MC were studied by the Optimal Docking Area (ODA) method to evaluate protein-protein binding sites and to provide a basis for the identification of binding partners for this class of enzymes. The ODA method identifies surface patches with optimal desolvation energy based on the selection of low-energy docking regions, generated from a set of surface points around the protein. With few exceptions, the ODA method identified surface patches with a significant low-energy docking surface for all the MCPs with known three-dimensional structure. Overall, in 14 out of 24 cases, the detected ODA patches were correctly located (i.e. more than 50% of the predicted residues were in known protein-protein binding sites), yielding a global success rate of 58%. More specifically, the success rate increased up to 80% on the ODA patches detected for the catalytic domains of the M14A subfamily, independently on the partner. Interestingly, the ODA residues on the catalytic domain were correctly located in the interface with the N-terminal pro domain in all MCPs. The spatial distribution of the ODA patches for the different members of the family is in relation to the origin and function of the particular MCP, which allowed distinguishing between them. In good agreement with the experimentally characterized protein interfaces, the total average surface area of the theoretically derived ODA patches for the catalytic domain of MCPs is around 1700 A2 and their content in hydrophobic residues is about 40%. As a particular case, the average surface area of the ODA patches in MCPs of crop insect pests is about twice that of the MCPs of vertebrates, which might be related to their particular function. We recognized two binding regions for the catalytic domain of the MCPs, one of them accounting for nearly all the known intermolecular interactions made up by the enzymes. Protein inhibitors seem to have evolved to dock on this subset of ODA patches, evoking the binding mode of the N-terminal pro domains. The second binding region detected, for which no ligands have been identified so far, seems to be related to the acquisition/maintenance of the native structure of the peptidase. Overall, the ODA method has been successful in identifying low-energy docking areas in a set of structurally and functionally related proteins, suggesting that it can be easily extended to other families in the search for protein-protein binding sites and for their functional significance. PMID- 17407162 TI - Scoring function accuracy for membrane protein structure prediction. AB - We perform a systematic examination of the ability of several different high resolution, atomic-detail scoring functions to discriminate native conformations of loops in membrane proteins from non-native but physically reasonable, or "decoy," conformations. Decoys constructed from changing a loop conformation while keeping the remainder of the protein fixed are a challenging test of energy function accuracy. Nevertheless, the best of the energy functions we examined recognized the native structure as lowest in energy around half the time, and consistently chose it as a low-energy structure. This suggests that the best of present energy functions, even without a representation of the lipid bilayer, are of sufficient accuracy to give reasonable confidence in predictions of membrane protein structure. We also constructed homology models for each structure, using other known structures in the same protein family as templates. Homology models were constructed using several scoring functions and modeling programs, but with a comparable sampling effort for each procedure. Our results indicate that the quality of sequence alignment is probably the most important factor in model accuracy for sequence identity from 20-40%; one can expect a reasonably accurate model for membrane proteins when sequence identity is greater than 30%, in agreement with previous studies. Most errors are localized in loop regions, which tend to be found outside the lipid bilayer. For the most discriminative energy functions, it appears that errors are most likely due to lack of sufficient sampling, although it should be stressed that present energy functions are still far from perfectly reliable. PMID- 17407163 TI - RNASEL Arg462Gln polymorphism and prostate cancer in PLCO. AB - BACKGROUND: The Gln allele of the Arg462Gln polymorphism in RNASEL results in a threefold decrease in enzymatic activity, a reported deficiency in apoptotic response, and has been associated with prostate cancer in some high-risk family studies. The relationship of this variant to sporadic prostate cancer remains uncertain. METHODS: We conducted a nested case-control study of 1,317 prostate cancer cases and 1,842 controls from the screening arm of the prostate, lung, colorectal, and ovarian (PLCO) cancer screening trial. Conditional logistic regression was used to evaluate the association between the RNASEL Arg462Gln polymorphism and prostate cancer. RESULTS: No statistically significant association was observed between the Arg462Gln polymorphism and prostate cancer (compared to Arg/Arg, Gln/Arg: OR= 0.99 95% CI 0.84-1.16; Gln/Gln: OR= 0.95 95% CI 0.74-1.21), although slight non-significant differences in risk were observed among men with the Gln/Gln genotype by stage and grade. CONCLUSIONS: These results suggest that the RNASEL Gln/Gln genotype does not play an important role in the etiology of prostate cancer in the general population. PMID- 17407164 TI - Computational analysis of the S. cerevisiae proteome reveals the function and cellular localization of the least and most amyloidogenic proteins. AB - Protein sequences have evolved to optimize biological function that usually requires a well-defined three-dimensional structure and a monomeric (or oligomeric) state. These two requirements may be in conflict as the propensity for beta-sheet structure, which is one of the two most common regular conformations of the polypeptide chain in folded proteins, favors also the formation of ordered aggregates of multiple copies of the same protein (fibril, i.e., polymeric state). Such beta-aggregation is typical of amyloid diseases that include Alzheimer's, Parkinson's, and type II diabetes as well as the spongiform encephalopathies. Here, an analytical model previously developed for evaluating the amyloidogenic potential of polypeptides is applied to the proteome of the budding yeast (Saccharomyces cerevisiae). The model is based on the physicochemical properties that are relevant for beta-aggregation and requires only the protein sequence as input. It is shown that beta-aggregation prone proteins in yeast are accrued in molecular transport, protein biosynthesis, and cell wall organization processes while they are underrepresented in ribosome biogenesis, RNA metabolism, and vitamin metabolism. Furthermore, beta-aggregation prone proteins are much more abundant in the cell wall, endoplasmic reticulum, and plasma membrane than in the nucleolus, ribosome, and nucleus. Thus, this study indicates that evolution has not only prevented the selection of amyloidogenic sequences in cellular compartments characterized by a high concentration of unfolded proteins but also tried to exploit the beta-aggregated state for certain functions (e.g. molecular transport) and in well-confined cellular environments or organelles to protect the rest of the cell from toxic (pre-)fibrillar species. PMID- 17407165 TI - Dynamic alpha-helices: conformations that do not conform. AB - Structural transitions are important for the stability and function of proteins, but these phenomena are poorly understood. An extensive analysis of Protein Data Bank entries reveals 103 regions in proteins with a tendency to transform from helical to nonhelical conformation and vice versa. We find that these dynamic helices, unlike other helices, are depleted in hydrophobic residues. Furthermore, the dynamic helices have higher surface accessibility and conformational mobility (P-value = 3.35e-07) than the rigid helices. Contact analyses show that these transitions result from protein-ligand, protein-nucleic acid, and crystal contacts. The immediate structural environment differs quantitatively (P-value = 0.003) as well as qualitatively in the two alternate conformations. Often, dynamic helix experiences more contacts in its helical conformation than in the nonhelical counterpart (P-value = 0.001). There is differential preference for the type of short contacts observed in two conformational states. We also demonstrate that the regions in protein that can undergo such large conformational transitions can be predicted with a reasonable accuracy using logistic regression model of supervised learning. Our findings have implications in understanding the molecular basis of structural transitions that are coupled with binding and are important for the function and stability of the protein. Based on our observations, we propose that several functionally relevant regions on the protein surface can switch over their conformation from coil to helix and vice-versa, to regulate the recognition and binding of their partner and hence these may work as "molecular switches" in the proteins to regulate certain biological process. Our results supports the idea that protein structure-function paradigm should transform from static to a highly dynamic one. PMID- 17407166 TI - A model of restriction endonuclease MvaI in complex with DNA: a template for interpretation of experimental data and a guide for specificity engineering. AB - R.MvaI is a Type II restriction enzyme (REase), which specifically recognizes the pentanucleotide DNA sequence 5'-CCWGG-3' (W indicates A or T). It belongs to a family of enzymes, which recognize related sequences, including 5'-CCSGG-3' (S indicates G or C) in the case of R.BcnI, or 5'-CCNGG-3' (where N indicates any nucleoside) in the case of R.ScrFI. REases from this family hydrolyze the phosphodiester bond in the DNA between the 2nd and 3rd base in both strands, thereby generating a double strand break with 5'-protruding single nucleotides. So far, no crystal structures of REases with similar cleavage patterns have been solved. Characterization of sequence-structure-function relationships in this family would facilitate understanding of evolution of sequence specificity among REases and could aid in engineering of enzymes with new specificities. However, sequences of R.MvaI or its homologs show no significant similarity to any proteins with known structures, thus precluding straightforward comparative modeling. We used a fold recognition approach to identify a remote relationship between R.MvaI and the structure of DNA repair enzyme MutH, which belongs to the PD-(D/E)XK superfamily together with many other REases. We constructed a homology model of R.MvaI and used it to predict functionally important amino acid residues and the mode of interaction with the DNA. In particular, we predict that only one active site of R.MvaI interacts with the DNA target at a time, and the cleavage of both strands (5'-CCAGG-3' and 5'-CCTGG-3') is achieved by two independent catalytic events. The model is in good agreement with the available experimental data and will serve as a template for further analyses of R.MvaI, R.BcnI, R.ScrFI and other related enzymes. PMID- 17407167 TI - The impact of polyadenylation signals on plasmid nuclease-resistance and transgene expression. AB - BACKGROUND: Efficient delivery and expression of plasmids (pDNA) is a major concern in gene therapy and DNA vaccination using non-viral vectors. Besides the use of adjuvants, the pDNA vector itself can be designed to maximize survival in nuclease-rich environments. Homopurine-rich tracts in polyadenylation sequences have been previously shown to be especially important in pDNA resistance. METHODOLOGY: The effect of modifications in the poly A sequence of a model pDNA vector (pVAX1GFP) on nuclease resistance and transgene expression was investigated. Four poly A sequences were studied: bovine growth hormone (BGH), mutant BGH, SV40 and a synthetic poly A. Plasmid resistance (half-life) was assessed through in vitro incubations with mammalian nucleases. The impact in transgene expression was studied by quantifying pDNA, mRNA, and GFP expression in CHO, hybridoma and HeLa cells. RESULTS AND CONCLUSIONS: In vitro and cell culture studies indicate that plasmids containing the SV40 and the synthetic poly A sequences present significant improvements in nuclease resistance (up to two-fold increase in half-life). However, RT-PCR analysis demonstrated that significant reduction in mRNA steady-state levels were responsible for a decrease in transgene expression and detected transfection level of CHO and hybridoma cells when using the more resistant plasmids. Interestingly, transfection of HeLa cells demonstrated that both poly A efficiency and plasmid resistance interfere significantly in transgene expression. The results strongly suggest that the choice of the poly A is important, not only for mRNA maturation/stability, but also for pDNA resistance, and should thus be taken into consideration in the design and evaluation of pDNA vectors. PMID- 17407168 TI - The effect of chronic benzodiazepine use on cognitive functioning in older persons: good, bad or indifferent? AB - OBJECTIVE: This study investigates the effects of benzodiazepine (BZ) use on cognitive performance in elderly persons in a longitudinal design. STUDY DESIGN AND SETTING: Data were obtained from the Longitudinal Aging Study Amsterdam (LASA), in the Netherlands. 2,105 respondents (>/=62 years of age) were included and had repeated measurements over a period of 9 years. For all BZs the type, dosage, frequency and duration of use was measured. The equivalent of a dose of diazepam was determined with regard to type and dosage and a cumulative dosage was calculated. General cognitive functioning was measured with the Mini-Mental State Examination, information processing speed was measured with the coding task, fluid intelligence with Raven's Coloured Progressive Matrices and episodic memory with the Auditory Verbal Learning Test. Multilevel analyses were used to investigate the relationship between BZ use and cognitive decline. RESULTS: A negative effect of BZ use on cognitive performance was found. However, the effect sizes were very small. CONCLUSION: This study suggests that both duration and cumulative exposure to BZ has a small negative effect on the long-term cognitive functioning of elderly people in the community. PMID- 17407169 TI - What is elder abuse--who decides? AB - OBJECTIVES: To test the hypothesis that family and professional caregivers have different views about what constitutes elder abuse. DESIGN: A vignette was given to family and professional carers. They were asked to rate 13 management strategies for behavioural difficulties in a person with dementia on a Likert scale ranging from good idea to abusive. Some of the strategies were abusive according to the Department of Health's 'No Secrets' definition. SETTING AND PARTICIPANTS: Family carers were recruited as part of a study of Alzheimer's disease. We recruited professionals working in several clinical disciplines and settings from a mental health service for older people. RESULTS: Seventy-four family and 38 professional carers completed questionnaires. The only abusive strategy that significantly more professionals than family carers identified correctly was preventing someone moving by putting a table over their lap (33(86.8%) vs 35(47.3%), p < 0.0005). In contrast, significantly more family carers identified that the neglect item of accepting someone was not clean was abusive (21(28.4%) vs 3(7.9%), p = 0.009). CONCLUSIONS: Professionals and carers reported significantly different views from each other and guidelines about what constituted elder abuse. This may be because abuse remains unacknowledged if people feel there are no better management options, and reporting leads only to punitive action for the carer perhaps coupled with institutionalisation for the person with dementia. Successful guidelines require societal agreement about what constitutes abuse and that prevention leads to better outcomes. PMID- 17407170 TI - Caffeine and 35% carbon dioxide challenge tests in panic disorder. AB - Our aim was to compare the demographic and clinical features of panic disorder (PD) patients with agoraphobia-DSM-IV-who had a panic attack after both an oral caffeine and the 35% carbon dioxide (CO2) challenge tests (responsive group) and compare them with PD patients who did not have a panic attack after both tests (non-responsive group). We examined 83 PD patients submitted to a 35% CO2 test and to an oral caffeine (480 mg) intake within 1 week interval. A panic attack was induced in 51 (61.4%) patients during the CO2 test (chi2=31.67, df=1, p<0.001) and in 38 (45.8%) patients during the caffeine test (chi2=18.28, df=1, p=0.023). All patients who had a panic attack during the caffeine test also had a panic attack during the CO2 test (n=38)-responsive group. The responsive had more (chi2=24.55, df=1, p=0.008) respiratory PD subtype, disorder started earlier (Mann-Whitney, p<0.001) had a higher familial prevalence of PD (chi2=20.34, df=1, p=0.019), less previous alcohol abuse (chi2=23.42, df=1, p<0.001), and had more previous depressive episodes (chi2=27.35, df=1, p<0.001). Our data suggest that there is an association between respiratory PD subtype and hyperreactivity to challenge tests: CO2 and oral caffeine. PMID- 17407171 TI - Caregiver psychological characteristics predict discontinuation of care for disabled elderly at home. AB - OBJECTIVE: This study sought to determine the predictors of discontinuation of care for the disabled elderly at home using multivariate regression analysis. METHODS: In January to February 2005, using a self-administered questionnaire, data were collected from 193 caregivers of frail elderly listed on a roster for utilization of day service or short stay service from two Home Visit Nursing Care Stations. Family caregivers were defined as co-resident family members who provided a minimum of 1 h of daily care for at least 3 months. Multiple stepwise logistic regression analysis was performed to detect predictors of a discontinuation for the disabled elderly at home. RESULTS: The mean age of caregivers was around 59.0 years, accounting for about 80% of the women among caregivers. Of those caregivers, the proportion of desire to institutionalization (high DI) (56.8%) was higher than that of desire to care at home (low DI) (43.2%). Among the following three factors selected by multivariate stepwise logistic regression analysis, the strength of the relationship was stronger in frequent mood swings and irritability as carerecipient characteristic (adjusted OR = 5.93; 95% CI, 2.09-16.8) than in no advanced or skilled care (adjusted OR = 3.13; 95% CI, 1.41-7.14) and a high caregiver burden (adjusted OR = 1.12; 95% CI, 1.03-1.23) as caregiver's one. CONCLUSIONS: Our results suggest that the carerecipients' psychological characteristics may be more strongly related to the decision to continue care for disabled elderly at home than that of the caregivers'. PMID- 17407174 TI - Design, synthesis, and evaluation of tripeptidic promoieties targeting the intestinal peptide transporter hPEPT1. AB - The human intestinal proton coupled di/tri-peptide transporter hPEPT1 promotes the oral bioavailability of several drug compounds. The strategy behind the present work is that by linking a suitable di- or tripeptidic promoiety to a drug substance, by a hydrolysable ester bond, it may give rise to a prodrug that targets hPEPT1. 29 tripeptides were designed based on known structural requirements for substrates binding hPEPT1. Serine, homoserine, or threonine was incorporated in the tripeptide as hydroxy group donors in order for them to be linked to carboxylic drug substances. Optimisation of the promoiety included a study of 14 unnatural tripeptides whose diversity was expressed by VolSurf descriptors. A total of 29 tripeptides was synthesised by solid phase peptide synthesis and a standard Fmoc protocol. The affinity of the tripeptides to hPEPT1 was determined by measuring the inhibition of [(14)C]Gly-Sar in mature Caco-2 cell monolayers which resulted in K(i) values ranging from 0.22 to 25 mM or above. Translocation through the intestinal membrane, mediated by hPEPT1, was measured by recording the membrane potential relative to that induced by the known substrate Gly-Sar. The change in membrane potential is caused by influx of protons due to the co-transport of substrates and protons by hPEPT1 and is, as such, an indication of translocation. A K(i) value of 0.30 mM combined with efficient translocation indicated that H-Phe-Ser-Ala-OH is a suitable lead promoiety for targeted hPEPT1 prodrug design. PMID- 17407175 TI - The role of private providers in treating child diarrhoea in Latin America. AB - Diarrhoeal disease, a leading cause of child mortality, disproportionately affects children in low-income countries - where private and non-governmental providers are often an important source of health care. We use 10 Living Standards Measurement Surveys from Latin America to model the choice of care for child diarrhoea in the private sector compared to the public sector. A total of 36.8% of children in the combined data set saw a private provider rather than a public one when taken for treatment. Each additional quintile of household economic status is associated with an increase of 6.5 percentage points in the probability that a child with diarrhoea is taken to a private provider (p<0.001). However, treatments provided in the private sector are manifestly of worse quality than in the public sector. A total of 33.0% of children visiting a public provider received Oral Rehydration Solution, compared to 13.7% of those visiting a private provider. Conversely, children treated by a private provider are more likely to receive drugs, most commonly unnecessary antibiotics. Ironically, when it comes to treatment for child diarrhoea, wealthier and better educated households in Latin America are paying for treatment in the private sector that is ineffective in comparison with treatments that are commonly and inexpensively available. PMID- 17407176 TI - Caregiver preference for rivastigmine patch relative to capsules for treatment of probable Alzheimer's disease. AB - BACKGROUND: Family caregivers comprise a critical component in the care of Alzheimer's disease (AD) patients. Among their many tasks, caregivers are responsible for administering and managing medications. Effective interventions incorporate the needs of both the AD patient and the caregiver, and understanding treatment preferences may maximize intervention effectiveness. Transdermal patches may offer advantages over conventional oral formulations. METHODS: A 24 week randomized controlled trial compared the rivastigmine patch to the rivastigmine capsule and placebo in patients with probable AD. At baseline and Weeks 8 and 24, the AD Caregiver Preference Questionnaire (ADCPQ) was used to evaluate caregiver expectations, preferences and satisfaction with treatment. Double-dummy treatment blinding ensured that caregiver preference for the patch or capsule was not confounded by perceptions of efficacy or tolerability. Reasons for preference were also elicited. The analytic sample included caregivers who completed the ADCPQ at Weeks 8 and/or 24. RESULTS: One thousand and fifty-nine caregivers completed the ADCPQ. More than 70% of caregivers preferred the rivastigmine patch to the capsule. The patch was significantly preferred to the capsule with respect to ease of following the schedule and ease of use. Caregivers indicated greater satisfaction overall, greater satisfaction with administration, and less interference with daily life with the patch versus the capsule (all p 100-fold of wild-type (WT) levels but fell to background within 4 weeks post-injection. In immunomodulated transposon-treated MPS I mice plasma IDUA persisted for over 3 months at up to 100-fold WT activity in one-third of MPS I mice, which was sufficient to reverse lysosomal pathology in the liver and, partially, in distant organs. Histological and immunohistochemical examination of liver sections in IDUA transposon-treated WT mice revealed inflammation 10 days post-injection consisting predominantly of mononuclear cells, some of which were CD4- or CD8-positive. CONCLUSIONS: Our results demonstrate the feasibility of achieving prolonged expression of lysosomal enzymes in the liver and reversing MPS disease in adult mice with a single dose of therapeutic SB transposons. PMID- 17407191 TI - Virus recognition by specific natural antibodies and complement results in MHC I cross-presentation. AB - Natural antibodies (NAb) and complement (C') are important regulators of immune system activation. We have shown previously that the galactosyl-alpha1,3 galactosyl (Gal alpha1,3Gal) xenoantigen and the similar ABO histo-blood group antigens are transferred onto virus from the producer cell, resulting in sensitisation of the virus to the respective NAb in a C'-dependent manner. Here we show that measles virus (Mv) that expresses Gal alpha1,3Gal termini can drive the proliferation of human T cells in the presence of serum and autologous DC, whereas without such targets, measles, as expected, suppress T cell reactivity. The use of affinity-purified NAb to Gal alpha1,3Gal and rabbit C' demonstrated the components in human serum responsible for this effect. Proteasome inhibition and blocking of antigen presentation showed that the increased T cell proliferation was mediated by MHC class I cross-presentation of immune complexes. These results lend further support to the idea that polymorphic carbohydrates of the Gal alpha1,3Gal/ABO type serve as important targets for NAb and C' and that their expression on virus has influenced their evolution by contributing to protection against viral transmission within as well as between species. The adjuvance effect of this recognition, acting as a bridge between the natural innate and adaptive immune systems, also has important implications for vaccine development. PMID- 17407192 TI - NF-kappaB-regulated suppression of T-bet in T cells represses Th1 immune responses in pregnancy. AB - The molecular mechanisms that suppress Th1 immune responses in pregnancy are unknown. We assessed the expression of the Th1 cytokine transcription factor T bet. We isolated PBMC and T cells from non-pregnant and pregnant women and demonstrated that T-bet is specifically down-regulated in pregnancy under basal and stimulated conditions. Low levels of T-bet protein were detected in the nuclear fraction of unstimulated PBMC from non-pregnant, but not pregnant women. Nuclear levels of T-bet increased in response to PMA/ionomycin in PBMC from non pregnant, but not pregnant women. T-bet expression was greater in whole cell lysates of stimulated CD3(+) T cells from non-pregnant relative to pregnant women. NF-kappaB is specifically down-regulated in T cells in pregnant women, resulting in suppressed expression of Th1 cytokines IL-2, IFN-gamma and TNF alpha. In this study, down-regulation of NF-kappaB also resulted in diminished expression of T-bet. PMA induces NF-kappaB translocation, T-bet expression and IL 2, IFN-gamma and TNF-alpha production. Conversely, pre-incubation with SN50, and NF-kappaB oligodeoxyribonucleotide decoys suppressed PMA-induced NF-kappaB translocation and gene transcription, respectively, resulting in diminished T-bet expression and Th1 cytokine production. Therefore, maintenance of the cytokine environment for pregnancy success is mediated via strict regulation of Th1 immune responses, more specifically through control of NF-kappaB and T-bet transcription. PMID- 17407193 TI - Inhibition of NKG2D receptor function by antibody therapy attenuates transfer induced colitis in SCID mice. AB - A role for the activating NK-receptor NKG2D has been indicated in several autoimmune diseases in humans and in animal models of type 1 diabetes and multiple sclerosis, and treatment with monoclonal antibodies to NKG2D attenuated disease severity in these models. In an adoptive transfer-induced model of colitis, we found a significantly higher frequency of CD4(+)NKG2D(+) cells in blood, mesenteric lymph nodes, colon, and spleen from colitic mice compared to BALB/c donor-mice. We, therefore, wanted to study the effect of anti-NKG2D antibody (CX5) treatment initiated either before onset of colitis, when the colitis was mild, or when severe colitis was established. CX5 treatment decreased the detectable levels of cell-surface NKG2D and prophylactic administration of CX5 attenuated the development of colitis significantly, whereas a more moderate reduction in the severity of disease was observed after CX5 administration to mildly colitic animals. CX5 did not attenuate severe colitis. We conclude that the frequency of CD4(+)NKG2D(+) cells increase during development of experimental colitis. NKG2D may play a role in the early stages of colitis in this model, since early administration of CX5 attenuated disease severity. PMID- 17407194 TI - PPARgamma-dependent regulation of human macrophages in phagocytosis of apoptotic cells. AB - Macrophages acquire their capacity for efficient phagocytosis of apoptotic cells during their differentiation from monocytes. The peroxisome proliferator activated receptor gamma (PPARgamma) is highly up-regulated during this maturation program. We report that addition of PPARgamma antagonist during differentiation of human monocytes to macrophages significantly reduced the capacity of macrophages to engulf apoptotic neutrophils, but did not influence phagocytosis of opsonized bacteria. Macrophage-specific deletion of PPARgamma in mice also resulted in decreased uptake of apoptotic cells. The antagonist acted in a dose-dependent manner during the differentiation of human macrophages and could also reverse the previously observed augmentation of phagocytosis by glucocorticoids. Blocking activation of PPARgamma led to down-regulation of molecular elements (CD36, AXL, TG2 and PTX3) of the engulfment process. Inhibition of PPARgamma-dependent gene expression did not block the anti inflammatory effect of apoptotic neutrophils or synthetic glucocorticoid, but significantly decreased production of IL-10 induced by LPS. Our results suggest that during differentiation of macrophages natural ligands of PPARgamma are formed, regulating the expression of genes responsible for effective clearance of apoptotic cells and macrophage-mediated inflammatory responses. PMID- 17407195 TI - CD4-mediated functional activation of human CD4+CD25+ regulatory T cells. AB - Naturally occurring CD4(+)CD25(+)FoxP3(+) regulatory T cells (CD25(+) Tregs) constitute a specialized population of T cells that is essential for the maintenance of peripheral self-tolerance. The immune regulatory function of CD25(+) Tregs depends upon their activation. We found that anti-CD4 antibodies activate the suppressive function of human CD25(+) Tregs in a dose-dependent manner. We demonstrate that CD4-activated CD25(+) Tregs suppress the proliferation of CD4(+) and CD8(+) T cells, their IL-2 and IFN-gamma production as well as the capacity of CD8(+) T cells to re-express CD25. By contrast, anti CD4 stimulation did not induce suppressive activity in conventional CD4(+) T cells. These results identify CD4 as a trigger for the suppressive function of CD25(+) Tregs and suggest a possible CD4-mediated exploitation of these cells. PMID- 17407196 TI - T1/ST2 expression on Th2 cells negatively regulates allergic pulmonary inflammation. AB - The transmembrane form of T1/ST2 (ST2) is a specific marker on murine Th2 cells that have been generated in vitro, or isolated from sites of allergic type 2 inflammation. Despite the association of ST2 with Th2 cells, to date no obligate role for ST2 in type 2 responses in vivo has been described. We have specifically addressed the role of ST2 on T cells by generation of ST2(-/-) mice crossed with ovalbumin (OVA) T cell receptor-transgenic mice. OVA-specific ST2(-/-) cells had normal cytokine responses to T cell activation after in vitro Th2 differentiation, but OVA stimulation of IL-5 was increased. Transfer of OVA specific ST2(-/-) Th2 cells into BALB/c mice caused exacerbated pulmonary inflammation with occluded airways, elevated airway hyper-responsiveness and increased susceptibility to methacholine challenge that was associated with mortalities of recipient mice. The increased pulmonary inflammation in OVA specific ST2(-/-) Th2 cell recipients was associated with selective differences in pulmonary levels of Th2 cytokines compared with OVA-specific ST2(+) Th2 cell recipients. Recipients of OVA-specific ST2(-/-) Th2 cells had a significant increase in eosinophils and a significant reduction in F4/80(hi) macrophages in the lungs. This is the first demonstration of a role for ST2 expression on Th2 cells down-regulating pulmonary inflammation. These data have major implications for the targeting of ST2 as a therapy for allergic airway disorders. PMID- 17407197 TI - Development of a method for the quantification of the molar gold concentration in tumour cells exposed to gold-containing drugs. AB - The knowledge of the cellular molar concentration of a drug is an extremely important parameter for the discussion and interpretation of its efficacy and bioavailability. Concerning metal complexes, electrothermal atomic absorption spectroscopy (ETAAS) offers a valuable analytical tool. However, matrix effects often hamper proper quantification of the metal concentration in biological tissues. This paper describes the development of an ETAAS method for the quantification of the molar gold concentration in HT-29 colon carcinoma cells. ETAAS analytical conditions were optimised and a factor was developed which allows the calculation of the molar cellular gold concentration from the measured gold per cellular biomass value. The method was used to quantify the gold content in HT-29 cells after exposure to the gold drug auranofin. Results indicated a strong cellular uptake of auranofin (compared to other metal anticancer drugs), which significantly correlated with the antiproliferative effects triggered by this agent. PMID- 17407198 TI - Bayesian generalized monotonic functional mixed models for the effects of radiation dose histograms on normal tissue complications. AB - When treating cancer patients with radiation therapy, the normal tissue in an organ close to the tumour usually receives some dose of radiation. The dose is not of the same intensity throughout the organ. This radiation can cause normal tissue complications, so for treatment planning purposes, it is important to understand the relationship between the distribution of dose intensities in the organ and the occurrence of complications. One general summary measure of the dose effect is obtained by integrating a weighting function (w(d)) over the dose distribution. The weighting function w(d) should be monotone for biological reasons. Because the true shape of w(d) is not known, we estimate it non parametrically subject to the monotonicity constraint. In our approach w(d) is written as a weighted sum of monotone basis functions. The weights in this sum are formulated as a mixture of point mass at zero and a Gamma random variable. A key feature of our method is that it allows for flat regions through the use of this mixture prior. The model is estimated using a Markov Chain Monte Carlo algorithm. We illustrate our method with data from a head and neck cancer study in which the irradiation of the parotid gland results in loss of saliva flow. PMID- 17407199 TI - Stoichiometry of biological nitrogen transformations in wetlands and other ecosystems. AB - The stoichiometric equations of ammonification, nitrification and denitrification have demonstrated that the nitrogen cycle in nature is rather complicated. The mechanisms of biological nitrogen transformations are very important for analysis, design, operation and optimal control of natural ecosystems or engineered systems for nitrogen removal, and accurate stoichiometric equations can help in the maintenance of these environments. In this study, the new stoichiometric equations of intermediate nitrification, and heterotrophic and autotrophic denitrification with sulfur as the electron donor have been developed and discussed. The parameter values of f(s) (the fraction of electron donor coupled to cell synthesis) in stoichiometric equations of nitrification and denitrification are calculated according to experimental results implied in previously reported stoichiometric equations. Some new stoichiometric relationships of nitrification and denitrification, such as the O(2) demand for nitrifications, chemical oxygen demand/N ratios and the yield coefficients for denitrifications have been established. The pathway steps of nitrification and denitrification have been discussed. PMID- 17407203 TI - Editorial: perspectives on milk--a special kind of food. PMID- 17407205 TI - Report: EuroBioFund first annual forum in Helsinki. PMID- 17407206 TI - Focus on: New Zealand. PMID- 17407207 TI - Reversing symptoms in autistic disorder. PMID- 17407208 TI - Freely accessible results of industrial genome study. PMID- 17407209 TI - Development of a stem cell tool box. PMID- 17407210 TI - Putting microbes to work: dairy fermentation, cell factories and bioactive peptides. Part I: overview. AB - A variety of milk-derived biologically active peptides have been shown to exert both functional and physiological roles in vitro and in vivo, and because of this are of particular interest for food science and nutrition applications. Biological activities associated with such peptides include immunomodulatory, antibacterial, anti-hypertensive and opioid-like properties. Milk proteins are recognized as a primary source of bioactive peptides, which can be encrypted within the amino acid sequence of dairy proteins, requiring proteolysis for release and activation. Fermentation of milk proteins using the proteolytic systems of lactic acid bacteria (LAB) is an attractive approach for generation of functional foods enriched in bioactive peptides given the low cost and positive nutritional image associated with fermented milk drinks and yoghurt. In this review, we discuss the exploitation of such fermentation towards the development of functional foods conferring specific health benefits to the consumer beyond basic nutrition. In particular, in Part I, we focus on the release of encrypted bioactive peptides from a range of food protein sources, as well as the use of LAB as cell factories for the de novo generation of bioactivities. PMID- 17407211 TI - Putting microbes to work: dairy fermentation, cell factories and bioactive peptides. Part II: bioactive peptide functions. AB - A variety of milk-derived biologically active peptides have been shown to exert both functional and physiological roles in vitro and in vivo, and because of this are of particular interest for food science and nutrition applications. Biological activities associated with such peptides include immunomodulatory, antibacterial, anti-hypertensive and opioid-like properties. Milk proteins are recognized as a primary source of bioactive peptides, which can be encrypted within the amino acid sequence of dairy proteins, requiring proteolysis for release and activation. Fermentation of milk proteins using the proteolytic systems of lactic acid bacteria is an attractive approach for generation of functional foods enriched in bioactive peptides given the low cost and positive nutritional image associated with fermented milk drinks and yoghurt. In Part II of this review, we focus on examples of milk-derived bioactive peptides and their associated health benefits, to illustrate the potential of this area for the design and improvement of future functional foods. PMID- 17407213 TI - "Keep on truckin'" or "It's got you in this little vacuum": race-based perceptions in decision-making for total knee arthroplasty. AB - OBJECTIVE: Ethnic variation in the utilization of total knee arthroplasty (TKA) has been well documented, with African Americans showing significantly lower utilization rates. These differences are not explained by the prevalence or severity of knee osteoarthritis (OA). Patient preferences may contribute to differences in utilization since TKA is an elective surgery. We conducted a qualitative analysis of decision-making factors influencing ethnic preferences for TKA in patients with knee OA. METHODS: Six focus group discussions were conducted with participants being divided according to race: 2 African American groups, 2 Hispanic groups, and 2 Caucasian groups. Participants were all patients with knee OA attending the same treatment facility. A total of 37 patients participated, 14 male and 23 female. Group discussions were led by a race concordant facilitator. A grounded theory approach was used to analyze verbatim transcripts of the discussions. RESULTS: Thirty distinct categories of information emerged in the analysis of the data. Four of these categories showed variation among ethnic groups and included explanations of illness, changes in lifestyle, trust and skepticism, and paying for surgery. CONCLUSION: Patient attitudes and beliefs about TKA vary among ethnic groups. Differences in explanations of illness, changes in lifestyle, trust and skepticism, and paying for surgery are topics for discussion in the medical encounter. There is a need for open doctor-patient communication around individual experiences and beliefs in an effort to enhance decision-making for TKA. PMID- 17407214 TI - Prevention of glucocorticoid induced osteoporosis with alendronate or alfacalcidol: relations of change in bone mineral density, bone markers, and calcium homeostasis. AB - OBJECTIVE: To explore the relation of changes in measures of bone turnover and changes in bone mineral density (BMD) of the lumbar spine and total hip over 18 months in a double-blinded, randomized trial, comparing the effect of alfacalcidol (101 patients) versus alendronate (100 patients) on BMD in patients who recently started treatment with glucocorticoids for various rheumatic diseases. METHODS: Associations between changes in serum procollagen type I C propeptide (P1CP), fasting urine N-terminal telopeptide of type I collagen (NTx), serum calcium, parathyroid hormone (PTH), osteocalcin, and change from baseline in BMD over 18 months were explored with regression and correlation analyses. RESULTS: In both treatment groups, there was a statistically significant decrease in NTx. In the alfacalcidol group there was also a significant increase in P1CP and osteocalcin, in contrast to the alendronate group, but BMD in the alfacalcidol decreased versus an increase in the alendronate group (p < 0.001). In neither treatment group were changes in biochemical measures correlated with the change in BMD, with the exception of a negative correlation in the alendronate group between changes in total hip BMD and NTx. Use of alendronate resulted in an increased PTH in 27 patients, but the increase in BMD of these patients was not statistically significantly different compared to patients taking alendronate with normal PTH levels. CONCLUSION: Changes in BMD were not associated with changes in bone measures, with the exception of NTx in the alendronate group. For the patient taking glucocorticoids in clinical practice, the value of serial assessment of bone markers is low; changes in markers are no substitute for changes in BMD. ClinicalTrials.gov number: NCT00138983. PMID- 17407215 TI - A combination of 6 months of treatment with pyridostigmine and triweekly exercise fails to improve insulin-like growth factor-I levels in fibromyalgia, despite improvement in the acute growth hormone response to exercise. AB - OBJECTIVE: People with fibromyalgia (FM) often have low insulin-like growth factor-I (IGF-I) levels and a suboptimal growth hormone (GH) response to acute exercise. As previous work had demonstrated a normalization of the acute GH response to exercise with the use of pyridostigmine (PYD), we tested the hypothesis that 6 months of PYD therapy plus supervised exercise would increase IGF-I levels. METHODS: Subjects with primary FM were randomized into 4 groups: (1) PYD/exercise; (2) PYD/diet recall; (3) placebo/exercise; and (4) placebo/diet recall. The dosing of PYD was 60 mg tid for 6 months. Resting IGF-I levels were measured at baseline and after 6 months of treatment. In addition the acute GH response to exercise at VO2 max was measured at baseline and after treatment. RESULTS: A total of 165 FM subjects (mean age 49.5 yrs, 5 male) were entered and 154 (93.3%) completed the study. Six months of therapy (PYD plus exercise or exercise alone) failed to improve the IGF-I levels. The use of PYD 1 hour prior to exercise improved the acute GH response (4.54 ng/dl) compared to placebo (1.74 ng/dl) (p = 0.001) at the end of the 6-month trial. The acute GH response to exercise at baseline did not correlate with IGF-I, age, depression, medications, estrogen status, or obesity. CONCLUSION: A combination of triweekly supervised exercise plus the daily use of PYD for 6 months failed to increase IGF-I levels in patients with FM, despite the confirmation that PYD normalizes the acute GH response to strenuous aerobic exercise. PMID- 17407216 TI - Toll-like receptor in salivary glands from patients with Sjogren's syndrome: functional analysis by human salivary gland cell line. AB - OBJECTIVE: We investigated expression of toll-like receptor (TLR) in labial salivary glands of patients with Sjogren's syndrome (SS) and functional TLR expression in the cultured salivary gland cell line. METHODS: Expression of TLR2, TLR3, TLR4, and myeloid differentiation factor 88 (MyD88) in labial salivary glands was examined by immunohistochemistry. Human salivary gland (HSG) cell-line cells were cultured with TLR ligands [peptidoglycan, poly (I:C) and lipopolysaccharide], and CD54 expression and interleukin 6 (IL-6) production was studied. Phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N terminal kinase (JNK), p38, and Akt was examined by Western blotting. Activation of nuclear factor-kappaB (NF-kappaB) p65 in HSG cells was studied by NF-kappaB p65 nuclear translocation by microscopic immunofluorescence or chemiluminescent electrophoretic mobility shift assay and detection of NF-kappaB p65 phosphorylation. RESULTS: TLR2, TLR3, TLR4, and MyD88 were more strongly expressed in the labial salivary glands of SS patients (n =12) than in control subjects (n = 4), and were found in salivary-infiltrating mononuclear cells as well as acinar cells and ductal epithelial cells. In cultured HSG cells, a similar expression pattern was observed, and TLR ligands stimulated CD54 expression and IL-6 production. TLR ligands induced phosphorylation of ERK, JNK, and p38 in HSG cells, but not Akt phosphorylation or activation of NF-kappaB p65. CONCLUSION: Although the putative ligands remain to be determined, our study indicated the activation of the TLR-mediated immune response in SS, and suggested that the TLR effect is mediated through the mitogen-activated protein kinase pathway. PMID- 17407217 TI - Soluble CD154 is not associated with atherosclerosis in systemic lupus erythematosus. AB - OBJECTIVE: Soluble CD154 (sCD154) is involved in the pathogenesis of systemic lupus erythematosus (SLE), as well as in the initiation and progression of atherosclerotic lesions. We determined the association of sCD154 with coronary calcium and carotid plaque at the baseline visit of the Lupus Atherosclerosis Prevention Study. METHODS: Serum samples were assayed for soluble CD154 by ELISA. Coronary calcium was measured by helical computed tomography. Carotid duplex was performed to measure carotid plaque. RESULTS: sCD154 was measured in 183 patients with SLE. Patients had a mean age of 48.8 +/- 10.5 yrs, and 92% were female. Ethnicity included 61% Caucasian, 34% African American, 2% Asian, and 2% Hispanic. sCD154 was not associated with carotid plaque (p = 0.45) nor with coronary calcium (p = 0.43). Indeed, those with carotid plaque had a trend toward lower levels of sCD154 (474 +/- 29.2 vs 526 +/- 5 pg/ml; p = 0.45). CONCLUSION: sCD154 is not associated with subclinical measures of atherosclerosis in SLE, including carotid plaque and coronary calcium. PMID- 17407218 TI - Measurement of antinuclear antibodies by multiplex immunoassay: a prospective, multicenter clinical evaluation. AB - OBJECTIVE: We conducted a prospective, multicenter evaluation of autoantibody testing by multiplex immunoassay in patients with known or suspected connective tissue diseases (CTD). We evaluated agreement between multiplex immunoassay and enzyme immunoassay (EIA) and assessed the diagnostic utility of autoantibody profiles. METHODS: Samples from 908 patients with suspected CTD seen in rheumatology clinics were collected prospectively at 3 tertiary care centers. Diagnoses were established according to recognized classification criteria. Tests for autoantibodies were obtained by multiplex immunoassay and by EIA. The results of the multiplex immunoassay were analyzed using a previously validated interpretative algorithm, MDSS (Medical Decision Support Software), that suggests possible disease associations based on the pattern of results for the autoantibodies. RESULTS: The median patient age was 49.7 years; 83% were female. The most common diagnoses were rheumatoid arthritis in 352 patients and systemic lupus erythematosus (SLE) in 332 patients. Agreement between multiplex and EIA testing ranged from a high of 99% (95% CI 98% to 100%) for Jo-1 to a low of 79% (95% CI 76% to 82%) for antinuclear antibodies. The MDSS algorithm suggested an appropriate disease association in 75% to 100% of patients with SLE. The results varied depending on the disease and the autoantibodies present. CONCLUSION: These results suggest that patterns of autoantibodies detected by multiplex immunoassay testing, when analyzed by an interpretative algorithm, are useful in the evaluation of patients with CTD in situations of high disease prevalence. Further testing is necessary to determine its utility in settings of low disease prevalence. PMID- 17407219 TI - Incidence of postpartum thrombosis and preterm delivery in women with antiphospholipid antibodies and recurrent pregnancy loss. AB - OBJECTIVE: To determine the frequency of preterm deliveries and postpartum thrombotic events (TE) in pregnancies resulting in live birth in women with antiphospholipid antibodies (aPL) and a history of recurrent pregnancy loss (RPL) but without prior TE. METHODS: We reviewed the pregnancy outcomes of women referred to our clinic with a history of RPL. Prepregnancy investigation of RPL included history of TE and aPL positivity (anticardiolipin IgG and lupus anticoagulant). We recorded use of anticoagulation therapy during and after pregnancy, obstetric outcome, gestational age at delivery, and postpartum course. Included in our study were women with unexplained RPL with no history of TE attending our clinic who subsequently had pregnancies that resulted in a live birth. RESULTS: Over a 5-year period, 260 women with RPL and no history of TE had a live birth at our clinic. Eighty-seven (33.5%) were positive for aPL and 173 (66.5%) were negative for aPL. Twenty-four percent of deliveries in the aPL positive group occurred before 37 weeks' gestation compared to 9.8% of deliveries in the aPL-negative group (p = 0.004; 95% CI 0.052-0.234). There were no antepartum TE in either group. One woman in the aPL-positive group (1.1%) had a deep vein thrombosis 3.5 weeks postpartum while receiving prophylactic anticoagulant therapy, compared to none in the aPL-negative group. CONCLUSION: A significantly higher proportion of aPL-positive patients had preterm deliveries compared to aPL-negative patients, but pregnancy-related TE was infrequent: 99.0% of aPL-positive women with a history of RPL and no prior TE who had a live birth at our clinic had an uneventful pregnancy, delivery, and postpartum course. PMID- 17407220 TI - Consequences of rheumatoid arthritis for performance of social roles--a literature review. AB - OBJECTIVE: To obtain quantitative estimates of restrictions in participation, i.e., the performance of social roles, in patients with rheumatoid arthritis (RA). METHODS: Participation categories were selected from the International Classification of Functioning, Disability and Health (ICF) (preliminary) Comprehensive Core Set for RA. A literature search was performed utilizing PubMed and PsychInfo. Articles were included if: (1) performance in at least one of the participation categories was described; (2) patients with RA were compared to a healthy reference population or their performance over time was described; (3) published between 1995 and 2005; and (4) written in English. RESULTS: Seven participation categories were selected from the Comprehensive Core Set for RA, resulting in 50 articles included in the review. Almost all studies focused on remunerative employment (n = 30), recreation and leisure (n = 17), or both (n = 3). RA patients had an increased risk of being without a paid job compared to well adjusted reference groups (absolute difference 4% to 28%, odds ratios 1.2 to 3.4). Restrictions in employment occurred already within the early phase of RA and varied greatly among studies. Two years after diagnosis, disability benefits increased up to roughly 30% in some European cohorts. In the category of recreation and leisure most studies focused on socializing (n = 16). Patients with longstanding RA experienced a decrease in socializing (range, Cohen's d, 0.46 to -1.0), but changes over time were minor. CONCLUSION: RA patients experience restrictions in the performance of remunerative employment and in recreation and leisure (socializing). Due to the lack of studies, no conclusions on other ICF categories describing social roles could be made. PMID- 17407221 TI - Determining rheumatologists' accuracy at assessing functional disability in rheumatoid arthritis patients using the Health Assessment Questionnaire Disability Index. AB - OBJECTIVE: To test rheumatologists' accuracy in determining functional disability of their patients with rheumatoid arthritis (RA). METHODS: We used the Health Assessment Questionnaire-Disability Index (HAQ-DI) as our guide at assessing functional disability in patients with RA. Included were male and female patients, 18 to 65 years of age, diagnosed with RA. Demographic data collected included the patients' age, disease duration, rheumatoid factor (RF) status, presence of rheumatoid nodules, absence or presence of erosive disease, and class and stage of their disease. The primary endpoint was the mean difference in the patients' HAQ-DI scores versus that of the physicians' (mHAQ-Diff). Secondary endpoints were the mean difference in pain assessment scale (mPAS-Diff) score; and assessing to see if the physicians' HAQ-DI was altered by the patients' age, disease duration, RF status, rheumatoid nodules, absence or presence of erosive disease, and class or stage of the patient's RA. RESULTS: A total of 223 patients (139 female, 84 male) were evaluated. The mHAQ-Diff score was statistically significant at -0.3 (p = 0.03) with the rheumatologists more often overestimating the degree of functional disability in their RA patients. The mPAS-Diff score was 0.16, but this was not significant (p = 0.53). There was no significant difference between the scores based on sex, presence or absence of RF, erosions, or rheumatoid nodules. However, the rheumatologists' estimated HAQ and PAS did seem to be more accurate in patients with lower class and stage of their RA. CONCLUSION: Our results indicate that there is a clear difference between patients' and rheumatologists' assessment of patients' functional disability in RA, with the rheumatologists significantly overestimating the degree of this disability. Although the rheumatologists' accuracy at determining the amount of their patients' functional disability was poor overall, they were somewhat more accurate in patients with RA having less severe disease. PMID- 17407222 TI - Macrophage migration inhibitory factor upregulates angiogenic factors and correlates with clinical measures in rheumatoid arthritis. AB - OBJECTIVE: To investigate the relationship between macrophage migration inhibitory factor (MIF) levels and clinical measures in rheumatoid arthritis (RA), and the potential for regulation of angiogenesis in RA. METHODS: Serum and synovial fluid (SF) levels of MIF and vascular endothelial growth factor (VEGF) in patients with RA were determined by sandwich ELISA, and the relationships among MIF, VEGF, and RA clinical measures were analyzed. RA synovial fibroblasts were cultured with recombinant human MIF (rhMIF) and the production of VEGF and interleukin 8 (IL-8) were measured in the conditioned media. The angiogenic effect of MIF was examined using established measures of angiogenesis in vitro. RESULTS: Erythrocyte sedimentation rate, C-reactive protein, and the daily dosage of oral prednisolone were correlated with SF levels of MIF. The SF levels of MIF were found to be higher in patients with bony erosion than in those without (69.2 +/- 11.4 ng/ml vs 44.0 +/- 6.2 ng/ml; p = 0.045). MIF levels had good correlation with VEGF levels (r = 0.52, p < 0.001 in sera, and r = 0.6, p < 0.001 in SF). Production of the angiogenic factors VEGF and IL-8 was enhanced in cultured RA synovial fibroblasts stimulated by rhMIF. Endothelial tube formation was augmented when the endothelial cells were cultured with the conditioned media from rhMIF-pretreated SF mononuclear cells, and this phenomenon was reversed by anti-VEGF antibody. CONCLUSION: SF MIF may reflect the clinical activity in patients with RA, and rhMIF induces the angiogenic factors in RA synovial fibroblasts. These results suggest that MIF may be an important cytokine in the perpetuation of the angiogenic and inflammatory processes in patients with RA. PMID- 17407223 TI - Elective orthopedic surgery and perioperative DMARD management: many questions, fewer answers, and some opinions... PMID- 17407224 TI - All patients with rheumatoid arthritis should receive corticosteroids as part of their management. PMID- 17407225 TI - Studying the benefit/risk ratio of glucocorticoids in rheumatoid arthritis. PMID- 17407226 TI - The need to take a fresh look at criteria for hypermobility. PMID- 17407227 TI - Spinal mobility measures in spondyloarthritis: application of the OMERACT filter. PMID- 17407228 TI - Considering cardiovascular mortality in patients with rheumatoid arthritis from a different perspective: a role for autonomic dysregulation and obstructive sleep apnea. PMID- 17407229 TI - Cytochrome P450 polymorphism as a predictor of ovarian toxicity to pulse cyclophosphamide in systemic lupus erythematosus. AB - OBJECTIVE: Ovarian toxicity is a major concern with cyclophosphamide (CYC) therapy. CYC is a prodrug that is activated by cytochrome P450 (CYP) enzymes to its active metabolites that are responsible for ovarian toxicity. The amount of active metabolites produced depends on polymorphism in CYP 450 genes. We studied the association of CYP2C19 and CYP2B6 genetic polymorphism with ovarian toxicity in patients with systemic lupus erythematosus (SLE) treated with CYC. METHODS: Thirty-five patients with SLE who had exposure to CYC were genotyped for variant alleles of CYP2B6 and CYP2C19. Ovarian toxicity included ovarian insufficiency, defined as lack of menses for 4 months, and ovarian failure (premature menopause) as amenorrhea lasting > 12 months before the age of 45 years. RESULTS: The mean age at start of CYC was 24.5 + 8.5 years and the cumulative dose of CYC received was 9.3 +/- 2.8 g. At the time of study the median followup after CYC treatment was 3 (1-6) years. A total of 17 patients developed ovarian toxicity, of whom 11 patients had ovarian insufficiency and 6 had premature menopause. The frequencies of variant alleles CYP2B6*5 and CYP2C19*2 were 8.5% and 21%, respectively. Patients who were homozygous or heterozygous for variant allele CYP2C19*2 had a significantly lower risk of developing ovarian toxicity when compared to patients with wild-type allele CYP2C19*1 (3/13 vs 14/22; OR 0.136, 95% CI 0.028-0.653; p < 0.01). No association was seen with CYP2B6 polymorphism. CONCLUSION: Presence of the variant allele CYP2C19*2 is associated with lower risk of ovarian toxicity in Indian patients treated with CYC. PMID- 17407230 TI - Anti-dsDNA antibody assay: high specificity and sensitivity with a filtration radioassay in comparison to low specificity with the standard ELISA. AB - OBJECTIVE: To evaluate whether a new fluid-phase filtration radioassay possesses both high sensitivity and specificity compared with the currently used ELISA and Farr assays. METHODS: Sequential sera (25 samples) from 9 patients with systemic lupus erythematosus (SLE), sera from 20 patients with SLE possessing anti-dsDNA antibodies by the Crithidia assay, 75 patients with rheumatoid arthritis possessing rheumatoid factors, 50 healthy control subjects, 767 from patients with type 1 diabetes, and a commercial standard serum sample were tested for anti dsDNA antibodies with the 3 different assays. RESULTS: Of serial dilutions of a standard anti-dsDNA antibody sample, only the highest positive sample (50 IU/ml) in the ELISA and the highest 2 positive samples (50 and 25 IU/ml) in the Farr assay were above the normal range. In contrast, all dilutions (to 2.5 IU/ml) of the standard anti-dsDNA antibody sample were above the normal range in the filtration radioassay. Using the values of 50 healthy control subjects in each assay to define the normal range, all 25 sequential sera from 9 patients with SLE were positive. In addition, 20/20 of the SLE individual sera, 2/75 (2.7%) of the RA sera, and 12/767 (1.6%) of the diabetes sera were positive (signal above normal range) in the filtration radioassay. The SLE sera were further examined in 2 additional assays, ELISA and Farr assay, and both assays were less sensitive and specific compared with the filtration radioassay. CONCLUSION: The fluid-phase filtration radioassay demonstrated high sensitivity and specificity for the detection of anti-dsDNA antibodies in SLE, with the standard ELISA exhibiting lower specificity. We suggest that testing for anti-dsDNA antibodies can be improved using a fluid-phase filtration radioassay in comparison to commercial assays. PMID- 17407231 TI - Monocyte chemoattractant protein-1: plasma concentrations and A(-2518)G promoter polymorphism of its gene in systemic lupus erythematosus. AB - OBJECTIVE: To determine (1) whether the A(-2518)G polymorphism of CCL-2, the gene encoding monocyte chemoattractant protein-1 (MCP-1), is associated with disease, MCP-1 concentration, nephritis, or coronary artery calcification (CAC) in systemic lupus erythematosus (SLE); and (2) whether MCP-1 and homocysteine (Hcy) concentrations are correlated. METHODS: Statistical tests were applied to determine the relationships between CCL-2 A(-2518)G genotypes, plasma MCP-1 concentrations, and clinical variables in Caucasian and African American patients with SLE and controls. RESULTS: The CCL-2 (-2518)G allele was not significantly associated with SLE in the whole study sample (p = 0.07). Among Caucasians, but not African Americans, G allele carriers had significantly increased risk of SLE (OR 4.2, 95% CI 1.8-9.6, p < 0.0001). Genotype was not associated with nephritis, CAC, or MCP-1 concentrations when all patients or all controls were considered; however, among recently diagnosed patients, G allele carriers had significantly higher MCP-1 concentrations than AA homozygotes (p = 0.02). SLE patients had higher MCP-1 concentrations than controls (p < 0.0001), African American patients had higher concentrations than Caucasian patients (p = 0.006), and patients with nephritis had higher concentrations than those without nephritis (p = 0.02). Although not associated with CAC, MCP-1 concentrations were significantly positively correlated with Hcy. CONCLUSION. CCL-2 A(-2518)G genotype is a significant risk factor for SLE among Caucasians but not African Americans, suggesting that genetically mandated differences in MCP-1 expression contribute to SLE etiology in the former. The positive correlation between MCP-1 and Hcy concentrations is consistent with the hypothesis that active inflammation and hyperhomocysteinemia are etiologically linked. PMID- 17407232 TI - Analysis of gene expression patterns in systemic sclerosis fibroblasts using RNA arbitrarily primed-polymerase chain reaction for differential display. AB - OBJECTIVE: To identify genes that are differentially expressed in systemic sclerosis (SSc) fibroblasts of clinically involved and noninvolved skin compared to normal dermal fibroblasts, using RNA arbitrarily primed-polymerase chain reaction (RAP-PCR) for differential display. METHODS: We examined 12 fibroblast cultures derived from clinically involved skin, 3 fibroblast cultures from noninvolved skin, and 4 fibroblast cultures from healthy skin. After extraction of total RNA, the first step of RAP-PCR was performed using different arbitrary 10-12-base primers for first-strand cDNA synthesis. Second-strand synthesis was achieved by cycling using different arbitrary 10-base primers, followed by sequence analysis of the amplified fingerprint products. The resulting sequences were aligned to the GenBank database using Blast Search. Confirmation of differential expression was performed with specific primers using real-time PCR. RESULTS: Using 8 different primer combinations, in total 48 cDNA were differentially expressed between SSc and healthy dermal fibroblasts. Sequence analysis identified distinct PCR products, which were overexpressed in SSc as highly homologous to gene segments of gremlin protein, lysyl oxidase, c-cbl proto oncogene, an estrogen-responsive element, fibronectin, and collagen type XIIa1 precursor. CONCLUSION: Our results show that RAP-PCR is a suitable method to identify differentially expressed genes in SSc fibroblasts. Further, we identified genes that have not yet been described in the pathophysiology of SSc and that may be involved in matrix synthesis and cellular interaction. PMID- 17407234 TI - Acitretin-related ossification. PMID- 17407233 TI - Epidemiology of general joint hypermobility and basis for the proposed criteria for benign joint hypermobility syndrome: review of the literature. AB - OBJECTIVE: This literature review of generalized joint hypermobility (GJH) syndromes discusses information regarding sex-, age-, and race-related factors from publications that specifically document validated GJH criteria. METHODS: We present an analysis of criterion-referenced connections that identify similarities among major and minor clinical criteria that identify both GJH and benign joint hypermobility syndrome (BJHS). In our search, we found considerable empirical evidence that supports an increased prevalence of hypermobility among children, women, and certain racial groups. Two commonly used clinical assessment tools, the Carter and Wilkinson criteria (>or= 3 positive tests out of 5) and the Beighton method (>or= 4 positive tests out of 9), are the sources of these data. BJHS is diagnosed through a set of major and minor criteria - a combination of symptoms and objective findings -- that include arthralgia, back pain, spondylosis, spondylolysis/spondylolisthesis, joint dislocation/subluxation, soft tissue rheumatism, marfanoid habitus, abnormal skin, eye signs, varicose veins or hernia or uterine/rectal prolapse. RESULTS: Clinically, there is some evidence that arthralgia, the proposed BJHS major criterion, is a major component of alleged hypermobility-related problems. In contrasting, there is no clear evidence that proposed BJHS minor diagnostic criteria are associated with hypermobility-related problems. An empirical correlation between hypermobility and osteoarthritis is possible, but so far unproven. There are no randomized controlled studies regarding effects of existing treatments. CONCLUSION: Generalized hypermobility is both sex- and age-related. Racial differences are also identifiable. The existence of BJHS can be accepted using present criteria. PMID- 17407235 TI - Summary findings of a systematic review of the ultrasound assessment of synovitis. AB - This report presents the results of a recent systematic review performed by the OMERACT Ultrasound Group on the metric properties of ultrasound for the detection of synovitis in inflammatory arthritis. Reviews were conducted for the hand, wrist, elbow, shoulder, knee, ankle, and foot; most reports were related to the hand and knee, and the most common disease process was rheumatoid arthritis. The review highlights the current gaps in the literature, including a lack of reliability data with respect to intra-occasion and intra- and inter-reader reliability. Current work by our group is addressing these issues. PMID- 17407236 TI - The OMERACT Ultrasound Group: status of current activities and research directions. AB - Ultrasound (US) is a relatively new imaging modality in rheumatology that offers great potential as a diagnostic and management tool. In 2004, an OMERACT Ultrasound Special Interest Group was formed to address the metric qualities of US as a potential outcome measure. A preliminary systematic review highlighted the deficiencies in the literature, particularly with regard to the reliability of interpreting and acquiring images; as a consequence, a number of exercises were proposed to address these issues. This report describes a series of iterative studies that have resulted in improved intra- and inter-reader reliability for detecting and scoring synovitis from both static and real-time images of the hand joints of patients with rheumatoid arthritis. The reliability of acquiring images was also enhanced using standardized positions. Future studies will assess the value of US in clinical trials. PMID- 17407237 TI - The OMERACT Magnetic Resonance Imaging Inflammatory Arthritis Group - advances and priorities. AB - This article updates the work and research priorities of the OMERACT working group on magnetic resonance imaging (MRI) in inflammatory arthritis, as presented to the OMERACT 8 meeting in Malta in May 2006. This work focused on testing the reliability of dedicated extremity MRI in rheumatoid arthritis and on the initial steps in the development of an MRI score for peripheral psoriatic arthritis. PMID- 17407238 TI - A multireader reliability study comparing conventional high-field magnetic resonance imaging with extremity low-field MRI in rheumatoid arthritis. AB - The use of extremity low-field magnetic resonance imaging (E-MRI) is increasing, but relatively few data exist on its reproducibility and accuracy in comparison with high-field MRI, especially for multiple readers. The aim of this multireader exercise of rheumatoid arthritis wrist and metacarpophalangeal joints was to assess the intermachine (high vs low-field) agreement and to assess the interreader agreement on high and low-field images. Study findings suggested that E-MRI performs similarly to conventional high-field MRI regarding assessment of bone erosions. However, for synovitis and bone edema, considerable intermachine and interreader variability was found. Further studies are needed before recommendations on multireader E-MRI assessment of these pathologies can be given. PMID- 17407239 TI - A multicenter reliability study of extremity-magnetic resonance imaging in the longitudinal evaluation of rheumatoid arthritis. AB - There are limited data on the reliability of extremity magnetic resonance imaging (E-MRI) in the longitudinal evaluation of rheumatoid arthritis (RA). Our aim was to assess the interreader reliability of the OMERACT RA MRI score in the assessment of change in disease activity and bone erosion scores using 0.2 T E MRI hand and wrist images from 2 timepoints, evaluated by 3 readers at different international centers. The intraclass correlation coefficients and smallest detectable difference results for the change scores were generally good for erosions and synovitis, but were not acceptable for bone edema. Overall, E-MRI demonstrated ability to detect change comparable to that reported for high-field MRI for erosion and synovitis. PMID- 17407240 TI - Developing a magnetic resonance imaging scoring system for peripheral psoriatic arthritis. AB - We describe the first steps in developing an OMERACT magnetic resonance imaging (MRI) scoring system for peripheral psoriatic arthritis (PsA). A preexisting MRI dataset (finger joints) from 10 patients with PsA was scored by 4 readers for bone erosion, bone edema, synovitis, tendinopathy, and extracapsular features of inflammation (including enthesitis) according to specified criteria. Scoring reliability between readers was moderate to high for bone edema and erosion, but lower for soft tissue inflammation. Measures to improve reliability for future exercises will include reviewing definitions of pathological features and prior reader calibration. PMID- 17407241 TI - Scoring inflammatory activity of the spine by magnetic resonance imaging in ankylosing spondylitis: a multireader experiment. AB - OBJECTIVE: Magnetic resonance imaging (MRI) of the spine is increasingly important in the assessment of inflammatory activity in clinical trials with patients with ankylosing spondylitis (AS). We investigated feasibility, inter reader reliability, sensitivity to change, and discriminatory ability of 3 different scoring methods for MRI activity and change in activity of the spine in patients with AS. METHODS: Thirty sets of spinal MRI at baseline and after 24 weeks of followup, derived from a randomized clinical trial comparing a tumor necrosis factor (TNF)-blocking drug (n = 20) with placebo (n = 10) and selected to cover a wide range of activity at baseline and change in activity, were presented electronically in a partial latin-square design to 9 experienced readers from different countries (Europe, Canada). Readers scored each set of MRI 3 times, using 3 different methods including the Ankylosing Spondylitis spine Magnetic Resonance Imaging-activity [ASspiMRI-a, grading activity (0-6) per vertebral unit in 23 units]; the Berlin modification of the ASspiMRI-a; and the Spondyloarthritis Research Consortium of Canada (SPARCC) scoring system, which scores the 6 vertebral units considered by the reader as the most abnormal, with additional scores for "depth" and "intensity." Both the order of the methods used by each reader and the timepoints (before/after treatment) were randomized. Feasibility of each scoring system was evaluated by measuring the mean time needed to score each set of MRI, and inter-reader reliability was evaluated by smallest detectable change (SDC) and by intraclass correlation coefficients (ICC) for all readers together and for all possible reader pairs separately. Sensitivity to change was investigated by calculating Guyatt's effect size on change scores. Discriminatory ability was assessed using Z-scores (Mann-Whitney test) comparing change in score between patients treated with TNF-blocking drug and placebo. RESULTS: The mean time to score one set of MRI was shortest for the Berlin method. SDC was lowest for the Berlin method and highest for SPARCC. Overall inter-reader ICC per method were between 0.49 and 0.77 for scoring activity status, and between 0.46 and 0.72 for scoring activity change. ICC for all possible reader pairs showed much more fluctuation per method, with lowest observed values of about 0.05 (very low agreement) and highest observed values over 0.90 (excellent agreement). In general, ICC for SPARCC were consistently higher than for other systems. Sensitivity to change differed per reader, and was more consistent with SPARCC than with the other methods, but was in general excellent for all 3 methods. Discrimination between groups (TNF-blocker vs placebo) assessed by Z-scores was good and comparable among methods. CONCLUSION: This experiment demonstrates the feasibility of multiple-reader MRI scoring exercises for method comparison, provides evidence for the feasibility, reliability, sensitivity to change, and discriminatory capacity of all 3 tested scoring systems to be used in assessing spinal activity on MRI in patients with AS in clinical trials. On the basis of these results it is not possible to prioritize one of the 3 methods. PMID- 17407242 TI - Is there a preferred method for scoring activity of the spine by magnetic resonance imaging in ankylosing spondylitis? AB - This report summarizes the discussion during a module update at OMERACT 8 on scoring methods for activity in the spine on magnetic resonance imaging. The conclusion was that the 3 available scoring methods are all very good with respect to discrimination and feasibility: the Ankylosing Spondylitis spine MRI score for activity (ASspiMRI-a), the Berlin method (a modification of the ASspiMRI-a), and the Spondyloarthritis Research Consortium of Canada Magnetic Resonance Imaging Index for Assessment of Spinal Inflammation in AS (SPARCC). All 3 methods were judged to be similar with respect to responsiveness and discrimination, although the differences in between-reader intraclass correlation coefficients (ICC) were judged to be relevant (the SPARCC method provided consistently higher ICC). The Berlin and SPARCC methods were preferred most frequently. The development of a new method combining the best elements of all methods is an additional possibility. PMID- 17407243 TI - Computer based methods for measurement of joint space width: update of an ongoing OMERACT project. AB - Computer-based methods of measuring joint space width (JSW) could potentially have advantages over scoring joint space narrowing, with regard to increased standardization, sensitivity, and reproducibility. In an early exercise, 4 different methods showed good agreement on measured change in JSW over time in the small joints of the hands and feet. Despite differences in measurement values between methods, measurement of within-joint change over time showed no systematic differences. The within-method variation was small, with intra operator variation being smaller than inter-operator variation. Although this initial study was limited in terms of the number of patients and timepoints (total 10), the number of joints was relatively high (340 joints), so the results were considered strong evidence supporting the validity of computer-based JSW measurements to continue the study of the potential value of JSW by comparison of measurements to manual scoring of joint space narrowing using the COBRA trial images. PMID- 17407244 TI - Repair in rheumatoid arthritis, current status. Report of a workshop at OMERACT 8. AB - Repair of structural damage in rheumatoid arthritis has drawn much attention with newly available effective treatments. A workshop was held at OMERACT 8 to update current knowledge on the validity of the concept of repair and on the assessment of repair. In preparation for the workshop several studies were performed and the results were presented. This was followed by a discussion and voting on statements on various aspects of repair. A majority of participants agreed that results of the new studies strengthen the validity of the concept of repair, and that repair can be assessed on radiographs. There was less agreement on the best means of measurement and there was a plea for more extensive reporting of data, i.e., not limited to sum scores of all joints together. The conclusions of the workshop mean a big step forward in the acceptance and assessment of repair. PMID- 17407245 TI - Seasonal variations in onset of Wegener's granulomatosis: increased in summer? PMID- 17407246 TI - Proceedings of the International Consensus Conference on Outcome Measures in Rheumatology Clinical Trails, OMERACT 8. May 10-14, 2006. St. Julian's Bay, Malta. PMID- 17407247 TI - Renal amyloidosis in rheumatoid arthritis. PMID- 17407248 TI - Preanalytical biases in measurement of matrix metalloproteinases and their tissue inhibitors in peripheral blood. PMID- 17407250 TI - Interleukin 1 receptor antagonist therapy-induced thrombocytopenia in adult onset Still's disease. PMID- 17407252 TI - Systemic lupus erythematosus with major organ involvement in a patient with systemic-onset juvenile idiopathic arthritis. PMID- 17407253 TI - The impact of IL-1Ra and MBL gene polymorphisms on joint damage after 5 and 10 years in patients with early rheumatoid arthritis. PMID- 17407254 TI - Obituary for an organization. PMID- 17407255 TI - Ivory vertebra and palmoplantar pustulosis. PMID- 17407256 TI - Antineutrophil cytoplasmic antibodies in nonsystemic vasculitic neuropathy. PMID- 17407257 TI - Refractory Wegener's meningitis treated with rituximab. PMID- 17407258 TI - Folic acid-conjugated protein cages of a plant virus: a novel delivery platform for doxorubicin. AB - The protein cage of a plant virus may provide a template for monodispersed nanosized systems for drug delivery. Using the Hibiscus chlorotic ringspot virus (HCRSV) as a model plant virus, we have prepared nanosized protein cages (30 nm) capable of encapsulating the anticancer drug, doxorubicin. The technique utilized the simultaneous encapsulation of a polyprotic acid of mw 200 kDa to produce an encapsulation efficiency for doxorubicin of about 7.5%. Folic acid was conjugated onto the capsids to impart cancer-targeting capability. The resultant nanosized systems improved the uptake and cytotoxicity of doxorubicin in the ovarian cancer cells, OVCAR-3, with statistical significance. Plant virus capsids may therefore provide viable templates for targeted drug delivery in cancer chemotherapy. PMID- 17407259 TI - Silicon nanowire arrays for label-free detection of DNA. AB - Arrays of highly ordered n-type silicon nanowires (SiNW) are fabricated using complementary metal-oxide semiconductor (CMOS) compatible technology, and their applications in biosensors are investigated. Peptide nucleic acid (PNA) capture probe-functionalized SiNW arrays show a concentration-dependent resistance change upon hybridization to complementary target DNA that is linear over a large dynamic range with a detection limit of 10 fM. As with other SiNW biosensing devices, the sensing mechanism can be understood in terms of the change in charge density at the SiNW surface after hybridization, the so-called "field effect". The SiNW array biosensor discriminates satisfactorily against mismatched target DNA. It is also able to monitor directly the DNA hybridization event in situ and in real time. The SiNW array biosensor described here is ultrasensitive, non radioactive, and more importantly, label-free, and is of particular importance to the development of gene expression profiling tools and point-of-care applications. PMID- 17407260 TI - A novel function for the N-terminal nucleophile hydrolase fold demonstrated by the structure of an archaeal inosine monophosphate cyclohydrolase. AB - Inosine 5'-monophosphate (IMP) cyclohydrolase catalyzes the cyclization of 5 formaminoimidazole-4-carboxamide ribonucleotide (FAICAR) to IMP in the final step of de novo purine biosynthesis. Two major types of this enzyme have been discovered to date: PurH in Bacteria and Eukarya and PurO in Archaea. The structure of the MTH1020 gene product from Methanothermobacter thermoautotrophicus was previously solved without functional annotation but shows high amino acid sequence similarity to other PurOs. We determined the crystal structure of the MTH1020 gene product in complex with either IMP or 5 aminoimidazole-4-carboxamide ribonucleotide (AICAR) at 2.0 and 2.6 A resolution, respectively. On the basis of the sequence analysis, ligand-bound structures, and biochemical data, MTH1020 is confirmed as an archaeal IMP cyclohydrolase, thus designated as MthPurO. MthPurO has a four-layered alphabeta betaalpha core structure, showing an N-terminal nucleophile (NTN) hydrolase fold. The active site is located at the deep pocket between two central beta-sheets and contains residues strictly conserved within PurOs. Comparisons of the two types of IMP cyclohydrolase, PurO and PurH, revealed that there are no similarities in sequence, structure, or the active site architecture, suggesting that they are evolutionarily not related to each other. The MjR31K mutant of PurO from Methanocaldococcus jannaschii showed 76% decreased activity and the MjE102Q mutation completely abolished enzymatic activity, suggesting that these highly conserved residues play critical roles in catalysis. Interestingly, green fluorescent protein (GFP), which has no structural homology to either PurO or PurH but catalyzes a similar intramolecular cyclohydrolase reaction required for chromophore maturation, utilizes Arg96 and Glu222 in a mechanism analogous to that of PurO. PMID- 17407261 TI - Determination of the three-dimensional structure of the Mrf2-DNA complex using paramagnetic spin labeling. AB - Understanding the mechanism of protein-DNA interactions at the molecular level is one of the main focuses in structural and molecular biological investigations. At present, NMR spectroscopy is the only approach that can provide atomic details of protein-DNA recognition in solution. However, determining the structures of protein-DNA complexes using NMR spectroscopy has been dependent on the observation of intermolecular nuclear Overhauser effects (NOE) and their assignments, which are difficult to obtain in many cases. In this study, we have shown that intermolecular distance constraints derived from a single spin-label in combination with docking calculations have defined many specific contacts of the complex between the AT-rich interaction domain (ARID) of Mrf2 and its target DNA. Mrf2 contacts DNA mainly using the two flexible loops, L1 and L2. While the L1 loop contacts the phosphate backbone, L2 and several residues in the adjacent helices interact with AT base pairs in the major groove of DNA. Despite the structural diversity in the ARID family of DNA-binding proteins, Mrf2 maintains contacts with DNA similar to those observed in the homologous Dri-DNA complex. PMID- 17407262 TI - Fatty acids covalently bound to alpha-hemolysin of Escherichia coli are involved in the molten globule conformation: implication of disordered regions in binding promiscuity. AB - Alpha-hemolysin (HlyA) is a pore-forming toxin secreted by pathogenic strains of Escherichia coli. The toxin is synthesized as a protoxin, ProHlyA, which is matured in the cytosol to the active form by acylation at two internal lysines, K563 and K689 (HlyA). It is widely known that the presence of fatty acids is crucial for the hemolytic and cytotoxic effects of the toxin. However, no detailed physicochemical characterization of the structural changes produced by fatty acids in the soluble protein prior to membrane binding has been carried out to date. The effects of chemical denaturants, the ANS binding parameters (Kd and n) and the sensitivity to proteases were compared between the acylated and unacylated protein forms HlyA and ProHlyA. Our results are consistent with a molten globular form of the acylated protein. Moreover, because molten globule proteins are intrinsically disordered proteins, using disorder prediction analyses, we show that HlyA contains 9 regions composed of 10-30 natively disordered amino acids. We propose that this conformation induced by covalently bound fatty acids might provide HlyA with the ability to bind to a variety of molecules during its action mechanism. PMID- 17407263 TI - A unique insert of leucyl-tRNA synthetase is required for aminoacylation and not amino acid editing. AB - Leucyl-tRNA synthetase (LeuRS) is a class I enzyme, which houses its aminoacylation active site in a canonical core that is defined by a Rossmann nucleotide binding fold. In addition, many LeuRSs bear a unique polypeptide insert comprised of about 50 amino acids located just upstream of the conserved KMSKS sequence. The role of this leucine-specific domain (LS-domain) remains undefined. We hypothesized that this domain may be important for substrate recognition in aminoacylation and/or amino acid editing. We carried out a series of deletion mutations and chimeric swaps within the leucine-specific domain of Escherichia coli. Our results support that the leucine-specific domain is critical for aminoacylation but not required for editing activity. Kinetic analysis determined that deletion of the LS-domain primarily impacts kcat. Because of its proximity to the aminoacylation active site, we propose that this domain interacts with the tRNA during amino acid activation and/or tRNA aminoacylation. Although the leucine-specific domain does not appear to be important to the editing complex, it remains possible that it aids the dynamic translocation process that moves tRNA from the aminoacylation to the editing complex. PMID- 17407264 TI - Human DNA ligase IV and the ligase IV/XRCC4 complex: analysis of nick ligation fidelity. AB - In addition to linking nicked/fragmented DNA molecules back into a contiguous duplex, DNA ligases also have the capacity to influence the accuracy of DNA repair pathways via their tolerance/intolerance of nicks containing mismatched base pairs. Although human DNA ligase I (Okazaki fragment processing) and the human DNA ligase III/XRCC1 complex (general DNA repair) have been shown to be relatively intolerant of nicks containing mismatched base pairs, the human DNA ligase IV/XRCC4 complex has not been studied in this regard. Ligase IV/XRCC4 is the sole DNA ligase involved in the repair of double strand breaks (DSBs) via the non-homologous end joining (NHEJ) pathway. During the repair of DSBs generated by chemical/physical damage as well as the repair of the programmed DSB intermediates of V(D)J recombination, there are scenarios where, at least conceptually, a capacity for ligating nicks containing mismatched base pairs would appear to be advantageous. Herein we examine whether ligase IV/XRCC4 can contribute a mismatched nick ligation activity to NHEJ. Toward this end, we (i) describe an E. coli-based coexpression system that provides relatively high yields of the ligase IV/XRCC4 complex, (ii) describe a unique rate-limiting step, which has bearing on how the complex is assayed, (iii) specifically analyze how XRCC4 influences ligase IV catalysis and substrate specificity, and (iv) probe the mismatch tolerance/intolerance of DNA ligase IV/XRCC4 via quantitative in vitro kinetic analyses. Analogous to most other DNA ligases, ligase IV/XRCC4 is shown to be fairly intolerant of nicks containing mismatched base pairs. These results are discussed in light of the biological roles of NHEJ. PMID- 17407265 TI - E. coli multidrug transporter MdfA is a monomer. AB - MdfA is a 410-residue-long secondary multidrug transporter from E. coli. Cells expressing MdfA from a multicopy plasmid exhibit resistance against a diverse group of toxic compounds, including neutral and cationic ones, because of active multidrug export. As a prerequisite for high-resolution structural studies and a better understanding of the mechanism of substrate recognition and translocation by MdfA, we investigated its biochemical properties and overall structural characteristics. To this end, we purified the beta-dodecyl maltopyranoside (DDM) solubilized protein using a 6-His tag and metal affinity chromatography, and size exclusion chromatography (SE-HPLC). Purified MdfA was analyzed for its DDM and phospholipid (PL) content, and tetraphenylphosphonium (TPP+)-binding activity. The results are consistent with MdfA being an active monomer in DDM solution. Furthermore, an investigation of two-dimensional crystals by electron crystallography and 3D reconstruction lent support to the notion that MdfA may also be monomeric in reconstituted proteoliposomes. PMID- 17407266 TI - Identification of acceptor substrate binding subsites +2 and +3 in the amylomaltase from Thermus thermophilus HB8. AB - Glycoside hydrolase family 77 (GH77) belongs to the alpha-amylase superfamily (Clan H) together with GH13 and GH70. GH77 enzymes are amylomaltases or 4-alpha glucanotransferases, involved in maltose metabolism in microorganisms and in starch biosynthesis in plants. Here we characterized the amylomaltase from the hyperthermophilic bacterium Thermus thermophilus HB8 (Tt AMase). Site-directed mutagenesis of the active site residues (Asp293, nucleophile; Glu340, general acid/base catalyst; Asp395, transition state stabilizer) shows that GH77 Tt AMase and GH13 enzymes share the same catalytic machinery. Quantification of the enzyme's transglycosylation and hydrolytic activities revealed that Tt AMase is among the most efficient 4-alpha-glucanotransferases in the alpha-amylase superfamily. The active site contains at least seven substrate binding sites, subsites -2 and +3 favoring substrate binding and subsites -3 and +2 not, in contrast to several GH13 enzymes in which subsite +2 contributes to oligosaccharide binding. A model of a maltoheptaose (G7) substrate bound to the enzyme was used to probe the details of the interactions of the substrate with the protein at acceptor subsites +2 and +3 by site-directed mutagenesis. Substitution of the fully conserved Asp249 with a Ser in subsite +2 reduced the activity 23-fold (for G7 as a substrate) to 385-fold (for maltotriose). Similar mutations reduced the activity of alpha-amylases only up to 10-fold. Thus, the characteristics of acceptor subsite +2 represent a main difference between GH13 amylases and GH77 amylomaltases. PMID- 17407267 TI - Equilibrium and kinetics of the folding and unfolding of canine milk lysozyme. AB - The equilibrium and kinetics of canine milk lysozyme folding/unfolding were studied by peptide and aromatic circular dichroism and tryptophan fluorescence spectroscopy. The Ca2+-free apo form of the protein exhibited a three-state equilibrium unfolding, in which the molten globule state is well populated as an unfolding intermediate. A rigorous analysis of holo protein unfolding, including the data from the kinetic refolding experiments, revealed that the holo protein also underwent three-state unfolding with the same molten globule intermediate. Although the observed kinetic refolding curves of both forms were single exponential, a burst-phase change in the peptide ellipticity was observed in both forms, and the burst-phase intermediates of both forms were identical to each other with respect to their stability, indicating that the intermediate does not bind Ca2+. This intermediate was also shown to be identical to the molten globule state observed at equilibrium. The phi-value analysis, based on the effect of Ca2+ on the folding and unfolding rate constants, showed that the Ca2+-binding site was not yet organized in the transition state of folding. A comparison of the result with that previously reported for alpha-lactalbumin indicated that the folding initiation site is different between canine milk lysozyme and alpha lactalbumin, and hence, the folding pathways must be different between the two proteins. These results thus provide an example of the phenomenon wherein proteins that are very homologous to each other take different folding pathways. It is also shown that the native state of the apo form is composed of at least two species that interconvert. PMID- 17407268 TI - Electrostatic and hydrophobic interactions governing the interaction and binding of beta-lactoglobulin to membranes. AB - The role of electrostatic and hydrophobic interactions in the binding and penetration of beta-lactoglobulin (betaLG) to preformed lipid membranes was studied using various phospholipid micelles and vesicles. Zwitterionic lysophospholipid micelles are able to induce the beta-sheet to alpha-helix transition, as judged by circular dichroism (CD), but the degree of transition is dramatically below and the amount of lipid required above that for anionic phospholipids with equivalent hydrocarbon chains. Anionic phospholipids with short hydrocarbon chains induce only low alpha-helical content in betaLG as compared to phospholipids with the same head group but longer hydrocarbon chains. These results suggest that both electrostatic and hydrophobic interactions are indispensable in betaLG-lipid interaction. Furthermore, air-water interface monolayer surface pressure and fluorescence anisotropy studies reveal that the membrane insertion of betaLG strongly depends on the nature of phospholipids, given the identical headgroup, particularly lipid packing. These results are supported by urea denaturation and acrylamide fluorescence quenching tests and by the FTIR-ATR polarization results for betaLG in multilayers on a surface. Under the same experimental conditions, the membrane binding and insertion of betaLG as well as the stability of the betaLG-lipid complexes can be enhanced by lowering the pH. Collectively, electrostatic interactions play a crucial role in all the processes involved in the betaLG-lipid interaction, while the presence of hydrophobic interaction remains necessary. Finally, betaLG biological function in the transport of fatty acids was tested by demonstrating the release of 2-AS from a 2-AS-betaLG complex on binding to lipids. PMID- 17407269 TI - The C-terminal domain of the archaeal leucyl-tRNA synthetase prevents misediting of isoleucyl-tRNA(Ile). AB - In the archaeal leucyl-tRNA synthetase (LeuRS), the C-terminal domain recognizes the long variable arm of tRNA(Leu) for aminoacylation, and the so-called editing domain deacylates incorrectly formed Ile-tRNA(Leu). We previously reported, for Pyrococcus horikoshii LeuRS, that a deletion mutant lacking the C-terminal domain (LeuRS_delta(811-967)) retains normal editing activity, but has severely reduced aminoacylation activity. In this study, we found that LeuRS_delta(811-967), but not the wild-type LeuRS, exhibited surprisingly robust deacylation activity against Ile-tRNA(Ile), correctly formed by isoleucyl-tRNA synthetase ("misediting"). Structural superposition of tRNA(Ile) onto the LeuRS x tRNA(Leu) complex indicated that Ile911, Lys912, and Glu913 of the LeuRS C-terminal domain clash with U20 of tRNA(Ile), which is bulged out as compared to the corresponding nucleotide of tRNA(Leu). The deletion of amino acid residues 911-913 of LeuRS enhanced the Ile-tRNA(Ile) deacylation activity, without affecting the Ile tRNA(Leu) deacylation activity. These results demonstrate that the clashing between U20 of tRNA(Ile) and residues 911-913 of the LeuRS C-terminal domain is the structural mechanism that prevents misediting. In contrast, the deletion of the C-terminal domains of the isoleucyl- and valyl-tRNA synthetases impaired both the aminoacylation (Ile-tRNA(Ile) and Val-tRNA(Val) formation, respectively) and editing (Val-tRNA(Ile) and Thr-tRNA(Val) deacylation, respectively) activities, and did not cause misediting (Val-tRNA(Val) and Thr-tRNA(Thr) deacylation, respectively) activity. Thus, the requirement of the C-terminal domain for misediting prevention is unique to LeuRS, which does not recognize the anticodon of the cognate tRNA, unlike the common aminoacyl-tRNA synthetases. PMID- 17407270 TI - Oligosilane chain-length dependence of electron transfer of zinc porphyrin oligosilane-fullerene molecules. AB - A new series of zinc porphyrin-fullerenes bridged by flexible oligosilane chains ZnP-[Sin]-C60 (n = 1-5) was synthesized, and the photophysical properties of these molecules were investigated using steady-state and time-resolved spectroscopic methods. The spectral observations can be well explained by assuming the coexistence of extended conformers and folded conformers, that is, the observed emissions are from the extended conformers while the folded conformers form very short lifetime nonfluorescent excited-state charge-transfer (CT) complexes. Time-resolved transient absorption spectra suggest the generation of intramolecular radical-ion pairs that have sub-microsecond lifetimes. With the number of silicon atoms of the bridged oligosilane, the lifetimes of the radical ion pairs do not vary regularly, indicating that intramolecular collision of the radical-cation moiety with the radical-anion moiety controls the charge recombination rate. The attenuation factor of the electron transfer of the silicon chain was evaluated by the bridge-length dependence of charge-separation rate to be 0.16 A-1 on the basis of the oligosilane chain-length dependence of fluorescence lifetimes. This is the first evaluation of the attenuation factor for the one-dimensional Si-Si chain to the best of our knowledge. PMID- 17407271 TI - Quenching of I(2P1/2) by O3 and O(3P). AB - Oxygen-iodine lasers that utilize electrical or microwave discharges to produce singlet oxygen are currently being developed. The discharge generators differ from conventional chemical singlet oxygen generators in that they produce significant amounts of atomic oxygen. Post-discharge chemistry includes channels that lead to the formation of ozone. Consequently, removal of I(2P1/2) by O atoms and O3 may impact the efficiency of discharge driven iodine lasers. In the present study, we have measured the rate constants for quenching of I(2P1/2) by O(3P) atoms and O3 using pulsed laser photolysis techniques. The rate constant for quenching by O3, (1.8 +/- 0.4) x 10(-12) cm3 s-1, was found to be a factor of 5 smaller than the literature value. The rate constant for quenching by O(3P) was (1.2 +/- 0.2) x 10(-11) cm3 s-1. PMID- 17407272 TI - Mid- and near-infrared spectra of conformers of H-Pro-Trp-OH. AB - We present near- and mid-infrared-UV double resonance spectra of the natural dipeptide H-Pro-Trp-OH. Two conformers are present in the supersonic expansion: a stretched conformer with fully extended backbone and a folded conformer with an OH...OCpep hydrogen bond. Both conformers are stabilized by dispersion interaction between indole ring and peptide backbone and a NHpep/Nproline contact. The vibrational and conformational assignment is supported by DFT and MP2 calculations. An adequate description of the energetic order of different conformers requires the explicit inclusion of dispersion and geometry optimization at the MP2 level. We will address the very sensitivity of the observed conformations to the structure of the end groups. PMID- 17407273 TI - Density functional theory study of the reaction mechanism and energetics of the reduction of hydrogen peroxide by ebselen, ebselen diselenide, and ebselen selenol. AB - Density functional theory calculations at the B3LYP/6-311++G(3df,3pd)//B3LYP/6 31G(d,p) level have been performed to elucidate the mechanism and reaction energetics for the reduction of hydrogen peroxide by ebselen, ebselen diselenide, ebselen selenol, and their sulfur analogues. The effects of solvation have been included with the CPCM model, and in the case of the selenol anion reaction, diffuse functions were used on heavy atoms for the geometry optimizations and thermochemical calculations. The topology of the electron density in each system was investigated using the quantum theory of atoms in molecules, and a detailed interpretation of the electronic charge and population data as well as the atomic energies is presented. Reaction free energy barriers for the oxidation of ebselen, ebselen diselenide, and ebselen selenol are 36.8, 38.4, and 32.5 kcal/mol, respectively, in good qualitative agreement with experiment. It is demonstrated that the oxidized selenium atom is significantly destabilized in all cases and that the exothermicity of the reactions is attributed to the peroxide oxygen atoms via reduction. The lower barrier to oxidation exhibited by the selenol is largely due to entropic effects in the reactant complex. PMID- 17407274 TI - New trans-platinum drugs with phosphines and amines as carrier ligands induce apoptosis in tumor cells resistant to cisplatin. AB - Cisplatin resistance observed in some human tumors has prompted research in platinum derivatives that can circumvent this effect. Despite initial works reporting lack of activity of trans-platinum derivatives, complexes with the general formula PtCl2(L)(L') exhibit cytotoxic activity in cisplatin-sensitive and -resistant cell lines. Here we reported the chemical and biological properties of seven platinum complexes with PPh3 or PMe2Ph in trans to several amines. They show important antitumoral properties in tumor cell lines. Among the compounds, those with a replacement of an ammine ligand in the inactive trans-DDP by a phosphine ligand have an important enhancement of their cytotoxic activity. In SKOV3, no G1 nor G2/M accumulation was observed after treatments, and apoptosis was launched probably by a mechanism independent of classical checkpoints activation. Our data indicate that our compounds are not cross resistant with cisplatin and might be promising agents in the treatment of tumors unresponsive to cisplatin. PMID- 17407275 TI - Structure-activity relationship of uridine 5'-diphosphoglucose analogues as agonists of the human P2Y14 receptor. AB - UDP-glucose (UDPG) and derivatives are naturally occurring agonists of the Gi protein-coupled P2Y14 receptor, which occurs in the immune system. We synthesized and characterized pharmacologically novel analogues of UDPG modified on the nucleobase, ribose, and glucose moieties, as the basis for designing novel ligands in conjunction with modeling. The recombinant human P2Y14 receptor expressed in COS-7 cells was coupled to phospholipase C through an engineered Galpha-q/i protein. Most modifications of the uracil or ribose moieties abolished activity; this is among the least permissive P2Y receptors. However, a 2 thiouracil modification in 15 (EC50 49 +/- 2 nM) enhanced the potency of UDPG (but not UDP-glucuronic acid) by 7-fold. 4-Thio analogue 13 was equipotent to UDPG, but S-alkylation was detrimental. Compound 15 was docked in a rhodposin based receptor homology model, which correctly predicted potent agonism of UDP fructose, UDP-mannose, and UDP-inositol. The hexose moiety of UDPG interacts with multiple H-bonding and charged residues and provides a fertile region for agonist modification. PMID- 17407276 TI - Pharmacological properties of bivalent ligands containing butorphan linked to nalbuphine, naltrexone, and naloxone at mu, delta, and kappa opioid receptors. AB - Our investigation of bivalent ligands at mu, delta, and kappa opioid receptors is focused on the preparation of ligands containing kappa agonist and mu agonist/antagonist pharmacophores at one end joined by a chain containing the mu antagonist pharmacophores (naltrexone, naloxone, or nalbuphine) at the other end. These ligands were evaluated in vitro by their binding affinity at mu, delta, and kappa opioid receptors and their relative efficacy in the [35S]GTPgammaS assay. PMID- 17407277 TI - Synthesis and pharmacological evaluation of a selected library of new potential anti-inflammatory agents bearing the gamma-hydroxybutenolide scaffold: a new class of inhibitors of prostanoid production through the selective modulation of microsomal prostaglandin E synthase-1 expression. AB - As a part of our drug discovery effort, recently we clarified the molecular basis of phospholipase A2 (PLA2) inactivation by petrosaspongiolide M (PM), an interesting metabolite belonging to a marine sesterterpene family, containing in its structural architecture a gamma-hydroxybutenolide moiety and showing potent anti-inflammatory activity. In the attempt to expand structural diversity as well as to simplify crucial synthetic features of the parent compound, we decided to develop a selected library based on the densely functionalized gamma hydroxybutenolide scaffold. The synthesized products were tested for their ability to inhibit PLA2 enzymes as well as to modulate the expression of inducible cyclooxygenase 2 (COX-2) and microsomal prostaglandin E synthase 1 (mPGES-1), two key enzymes highly involved in the inflammatory event, in order to discover new promising anti-inflammatory agents with better pharmacological profiles. This led us to the discovery of a promising inhibitor (4e) of prostanoid production acting by in vitro and in vivo selective modulation of microsomal prostaglandin E synthase 1 expression. PMID- 17407279 TI - Molecular properties investigation of a substituted aromatic mannich base: dynamic and static models. AB - An analysis of the hydrogen bridge of a Mannich base-type compound [3,5,6 trimethyl-2(N,N-dimethylaminomethyl)phenol, TMM] was performed according to the Car-Parrinello molecular dynamics (CPMD) scheme. A classical treatment of nuclei coupled with a first-principle potential energy surface was obtained from molecular dynamics simulation. Dipole moment values were collected during CPMD trajectory acquisition and subsequently used for the data analysis. The vibrational features and the intramolecular hydrogen-bond properties in the gas phase and solid state of the TMM compound were analyzed on the basis of widely used approaches: Fourier transformation of the autocorrelation function of both the atomic velocities and dipole moments. In addition, the time evolution of the structural parameters related to the hydrogen bond was carried out. The optimally localized Wannier functions served to describe the electronic structure of the Mannich base studied. The second part of the TMM compound study was performed in vacuo on the basis of density functional theory and second-order Moller-Plesset perturbation theory. The potential energy functions were used to solve the 1-D vibrational Schroedinger equation for the proton motion. This enabled a prediction of the anharmonic vibrational levels of the intramolecular hydrogen bond. The description of the electron density topology of the TMM molecule was carried out using the atoms in molecules theoretical framework. The computational results were further compared with the infrared spectra in the solid state. PMID- 17407280 TI - Ensemble feature selection: consistent descriptor subsets for multiple QSAR models. AB - Selecting a small subset of descriptors from a large pool to build a predictive quantitative structure-activity relationship (QSAR) model is an important step in the QSAR modeling process. In general, subset selection is very hard to solve, even approximately, with guaranteed performance bounds. Traditional approaches employ deterministic or stochastic methods to obtain a descriptor subset that leads to an optimal model of a single type (such as linear regression or a neural network). With the development of ensemble modeling approaches, multiple models of differing types are individually developed resulting in different descriptor subsets for each model type. However, it is advantageous, from the point of view of developing interpretable QSAR models, to have a single set of descriptors that can be used for different model types. In this paper, we describe an approach to the selection of a single, optimal, subset of descriptors for multiple model types. We apply this approach to three data sets, covering both regression and classification, and show that the constraint of forcing different model types to use the same set of descriptors does not lead to a significant loss in predictive ability for the individual models considered. In addition, interpretations of the individual models developed using this approach indicate that they encode similar structure-activity trends. PMID- 17407281 TI - From molecular to biological structure and back. AB - A comparative analysis of the topological structure of molecules and molecular biology networks revealed both similarity and differences in the methods used, as well as in the essential features of the two types of systems. Molecular graphs are static and, due to the limitations in atomic valence, show neither power distribution of vertex degrees nor "small-world" properties, which are typical for dynamic evolutionary networks. Areas of mutual benefits from an exchange of methods and ideas are outlined for the two fields. More specifically, chemical graph theory might make use of some new descriptors of network structure. Of interest for quantitative structure-property relationship/quantitative structure activity relationship and drug design might be the conclusion that descriptors based on distributions of vertex degrees, distances, and subgraphs seem to be more relevant to biological information than the single-number descriptors. The network concepts of centrality, clustering, and cliques provide a basis for similar studies in theoretical chemistry. The need of dynamic theory of molecular topology is advocated. PMID- 17407282 TI - Rare earth arenedisulfonate metal-organic frameworks: an approach toward polyhedral diversity and variety of functional compounds. AB - Eight 2D and 3D metal-organic framework (MOF) rare earth naphthalenedisulfonates have been obtained. The different geometry of the naphthalenedisulfonic acids used as connectors [(1,5-NDS) and (2,6-NDS)] gives rise to the three new structure types. In Ln(OH)(1,5-NDS)H2O, LnPF-1 (lanthanide polymeric framework; Ln=La, Nd, Pr, Sm and Eu), the lanthanide ion is octacoordinated. Its 3D structure is formed by (Ln2O14)-S-(Ln2O14) infinite chains, connected through complete NDS connectors. LnPF-2 (Ln=Nd), with the same empirical formula as the former, and the lanthanide in octa- and nonacoordination, owns an arrangement of sulfonate bridges and neodymium polyhedra that gives rise to a 2D structure. [Ln5(2,6-NDS)3(OH)9(H2O)4](H2O)2, LnPF-3 (Ln=Nd, Eu), demonstrates that it is possible to obtain a 3D structure with (2,6-NDS), when a greater Ln/connector ratio is employed. It is worth pointing out the existence, in this latter family of compounds, of a mu5-OH group, whose hydrogen atom is very close to one-sixth Ln atom (distance Ln...H=2.09 A). The materials, with high thermal stability, act as active and selective bifunctional heterogeneous catalysts in oxidation of linalool yielding cyclic hydroxy ethers. The absence of any 3D Nd-Nd magnetic interaction is explained due to the inner nature of 4f orbitals of Nd3+, which do not favor the magnetic exchange. The influence of the polymeric frame matrix results in a better photoluminescence efficiency for NdPF-1. PMID- 17407283 TI - The curium aqua ion. AB - The coordination environment of the hydrated Cm3+ ion is probed both in the solid state and in solution. The analysis of single-crystal X-ray diffraction data from [Cm(H2O)9](CF3SO3)3 determines that the Cm species is surrounded by nine coordinating waters with a tricapped-trigonal-prismatic geometry involving six short Cm-O distances at 2.453(1) A and three longer Cm-O distances at 2.545(1) A. The Cm nona-aqua triflate is isostructural with the series of lanthanide and actinide [R(H2O)9](CF3SO3)3 (R=La-Lu, Pu) compounds. A similar nona-aqua geometry is seen for the coordination environment of Cm in aqueous solution, as probed by high-energy X-ray scattering and extended X-ray absorption fine structure spectroscopy, although the splitting in the first coordination shell is increased from 0.092(2) in the solid to 0.16(2) A in solution. This increase in splitting of the Cm-water distances in the first coordination sphere is discussed in terms of its potential relevance to the previously observed decrease in coordinating waters with decreasing ionic radius about the f-ion in solution. PMID- 17407284 TI - DNA three-way junction with a dinuclear iron(II) supramolecular helicate at the center: a NMR structural study. AB - A tetracationic supramolecular helicate, [Fe2L3]4+ (L = C25H20N4), with a triple helical architecture is found to induce the formation of a three-way junction (3WJ) of deoxyribonucleotides with the helicate located in the center of the junction. NMR spectroscopic studies of the interaction between the M enantiomer of the helicate and two different oligonucleotides, [5'-d(TATGGTACCATA)]2 and [5' d(CGTACG)]2, show that, in each case, the 2-fold symmetry of the helicate is lifted, while the 3-fold symmetry around the helicate axis is retained. The 1:3 helicate/DNA stoichiometry estimated from 1D NMR spectra supports a molecular model of a three-way junction composed of three strands. Three separate double helical arms of the three-way junction are chemically identical giving rise to one set of proton resonances. The NOE contacts between the helicate and DNA unambiguously show that the helicate is fitted into the center of the three-way junction experiencing a hydrophobic 3-fold symmetric environment. Close stacking interactions between the ligand phenyl groups and the nucleotide bases are demonstrated through unusually large downfield shifts (1-2 ppm) of the phenyl protons. The unprecedented 3WJ arrangement observed in solution has also been found to exist in the crystal structure of the helicate adduct of [d(CGTACG)2] (Angew. Chem., Int. Ed. 2006, 45, 1227). PMID- 17407286 TI - Metal-organic framework based on a trinickel secondary building unit exhibiting gas-sorption hysteresis. AB - An interpenetrating microporous metal-organic framework based on a mu3 oxotrinickel basic carboxylate secondary building unit has been synthesized and structurally characterized. It exhibits pronounced gas-sorption hysteresis and selective adsorption of carbon dioxide over methane. PMID- 17407285 TI - X-ray structural characterization of imidazolylcobalamin and histidinylcobalamin: cobalamin models for aquacobalamin bound to the B12 transporter protein transcobalamin. AB - The X-ray structures of imidazolylcobalamin (ImCbl) and histidinylcobalamin (HisCbl) are reported. These structures are of interest given that the recent structures of human and bovine transcobalamin prepared in their holo forms from aquacobalamin show a histidine residue of the metalloprotein bound at the beta axial site of the cobalamin (Wuerges, J. et al. Proc. Natl. Acad. Sci. U.S.A. 2006, 103, 4386-4391). The beta-axial Co-N bond distances for ImCbl and HisCbl are 1.94(1) and 1.951(7) A, respectively. The alpha-axial Co-N bond distances to the 5,6-dimethylbenzimidazole are 2.01(1) and 1.979(8) A for ImCbl and HisCbl, respectively, and are typical for cobalamins with weak sigma-donor ligands at the beta-axial site. The corrin fold angles of 11.8(3) degrees (ImCbl) and 12.0(3) degrees (HisCbl) are smaller than those typically observed for cobalamins. PMID- 17407287 TI - Neutral organic mixed-valence compounds: synthesis and all-optical evaluation of electron-transfer parameters. AB - In this paper we present the synthesis as well as a detailed study of the electrochemical and photophysical properties of a series of neutral organic mixed valence (MV) compounds, 1-7, in which different amine donor centers are connected to perchlorinated triarylmethyl radical units by various spacers. We show that this new class of compounds are excellent model systems for the investigation of electron transfer due to their uncharged character and, consequently, their excellent solubility, particularly in nonpolar solvents. A detailed band shape analysis of the intervalence charge-transfer (IV-CT) bands in the context of Jortner's theory allowed the electron-transfer parameters (inner vibrational reorganization energy lambdav, outer solvent reorganization energy lambdao, and the difference in the free energy between the diabatic ground and excited states, DeltaG degrees degrees , as well as the averaged molecular vibrational mode v) to be extracted independently. In this way we were able to analyze the solvatochromic behavior of the IV-CT bands by evaluating the contribution of each parameter. By comparison of different compounds, we were also able to assign specific molecular moieties to changes in vv. For this class of molecules, we also demonstrate that the adiabatic dipole moment difference Deltamicroab and, consequently, the electronic coupling V12 can be evaluated directly from the absorption spectra by a new variant of the solvatochromic method. Furthermore, an investigation of the electrochemistry of compounds 1-7 by cyclic voltammetry as well as spectroelectrochemistry shows that, not only in the neutral MV compounds but also in their oxidized forms, a charge transfer can be optically induced but with exchanged donor-acceptor functionalities of the redox centers. PMID- 17407288 TI - Structural analysis of charge discrimination in the binding of inhibitors to human carbonic anhydrases I and II. AB - Despite the similarity in the active site pockets of carbonic anhydrase (CA) isozymes I and II, the binding affinities of benzenesulfonamide inhibitors are invariably higher with CA II as compared to CA I. To explore the structural basis of this molecular recognition phenomenon, we have designed and synthesized simple benzenesulfonamide inhibitors substituted at the para position with positively charged, negatively charged, and neutral functional groups, and we have determined the affinities and X-ray crystal structures of their enzyme complexes. The para-substituents are designed to bind in the midsection of the 15 A deep active site cleft, where interactions with enzyme residues and solvent molecules are possible. We find that a para-substituted positively charged amino group is more poorly tolerated in the active site of CA I compared with CA II. In contrast, a para-substituted negatively charged carboxylate substituent is tolerated equally well in the active sites of both CA isozymes. Notably, enzyme inhibitor affinity increases upon neutralization of inhibitor charged groups by amidation or esterification. These results inform the design of short molecular linkers connecting the benzenesulfonamide group and a para-substituted tail group in "two-prong" CA inhibitors: an optimal linker segment will be electronically neutral, yet capable of engaging in at least some hydrogen bond interactions with protein residues and/or solvent. Microcalorimetric data reveal that inhibitor binding to CA I is enthalpically less favorable and entropically more favorable than inhibitor binding to CA II. This contrasting behavior may arise in part from differences in active site desolvation and the conformational entropy of inhibitor binding to each isozyme active site. PMID- 17407289 TI - Uniformly aligned full-length membrane proteins in liquid crystalline bilayers for structural characterization. PMID- 17407290 TI - A meta effect in organic photochemistry? The case of SN1 reactions in methoxyphenyl derivatives. AB - The photochemistry of isomeric methoxyphenyl chlorides and phosphates has been examined in different solvents (and in the presence of benzene) and found to involve the triplet state. With the chlorides, C-Cl bond homolysis occurs in cyclohexane and is superseded by heterolysis in polar media, while the phosphate group is detached (heterolytically) only in polar solvents. Under such conditions, the isomeric triplet methoxyphenyl cations are the first formed intermediates from both precursors, but intersystem crossing (isc) to the singlets can take place. Solvent addition (forming the acetanilide in MeCN, the ethers in alcohols, overall a SN1 solvolysis) is a diagnostic reaction for the singlet cation, as reduction and trapping by benzene are for the corresponding triplet. Solvolysis is most important with the meta isomer, for which the singlet is calculated (UB3LYP/6-31 g(d)) to be the ground state of the cation (DeltaE = 4 kcal/mol) and isc is efficient (kisc ca. 1 x 108 s-1), and occurs to some extent with the para isomer (isoenergetic spin states, kisc ca. 1.7 x 106 s-1). The triplet is the ground state with the ortho isomer, and in that case isc does not compete, although trapping by benzene is slow because of the hindering of C1 by the substituent. The position of the substituent thus determines the energetic order of the cation spin states, in particular through the selective stabilization of the singlet by the m-methoxy group, a novel case of "meta effect". PMID- 17407291 TI - Trapping metal-organic framework nanocrystals: an in-situ time-resolved light scattering study on the crystal growth of MOF-5 in solution. PMID- 17407292 TI - A polyoxometalate-assisted electrochemical method for silicon nanostructures preparation: from quantum dots to nanowires. PMID- 17407295 TI - Photosensitization via dye coordination: a new strategy to synthesize metal nitrosyls that release NO under visible light. PMID- 17407296 TI - Extraordinarily high Zr4+ ion conducting solid. PMID- 17407297 TI - Oxidation of M4Si4 (M = Na, K) to clathrates by HCl or H2O. PMID- 17407298 TI - Highly stereoselective formal [3 + 3] cycloaddition of enals and azomethine imines catalyzed by N-heterocyclic carbenes. PMID- 17407299 TI - Cross-metathesis between alpha-methylene-gamma-butyrolactone and olefins: a dramatic additive effect. AB - [reaction: see text] Olefin cross-metathesis between alpha-methylene-gamma butyrolactone and terminal olefins is described. Moderate to excellent yields of alpha-alkylidene-gamma-butyrolactones were obtained with high E-stereoselectivity in the presence of low catalyst loading in refluxing CH2Cl2. In addition, the use of various additives was found to have a dramatic effect on the efficiency of the cross-metathesis (CM) process by circumventing the formation of the isomerized byproduct. PMID- 17407300 TI - Palladium-catalyzed annulation of aryl heterocycles with strained alkenes. AB - [reaction: see text] An annulation reaction proceeding by the intermolecular addition of an arylpalladium(II) halide across a strained alkene, followed by an intramolecular C-H functionalization of a pendant heterocycle is described. A variety of polycyclic heterocycles have been prepared from readily accessible haloaryl heterocycles by annulation with a range of strained alkene partners. PMID- 17407301 TI - Intramolecular 1,8- versus 1,6-hydrogen atom transfer between pyranose units in a (1-->4)-disaccharide model promoted by alkoxyl radicals. Conformational and stereochemical requirements. AB - [reaction: see text] The stereochemical and conformational factors controlling the intramolecular hydrogen atom transfer (HAT) reaction between the two pyranose units in a (1-->4)-disaccharide when promoted by a primary 6-O-yl radical are studied. Models with alpha-d-Glcp-(1-->4)-beta-d-Glcp or alpha-l-Rhamp-(1-->4) alpha-d-Galp skeletons lead exclusively to the abstraction of H-C-5' and the formation, through a nine-membered transition state, of a 1,3,5-trioxocane ring system in a stable boat-chair conformation. Notwithstanding, derivatives of alpha l-Rhamp-(1-->4)-alpha-d-Glcp abstract exclusively H-C-1' through a seven-membered transition state and therefore lead to an interglycosidic spiro ortho ester. PMID- 17407302 TI - Formal synthesis of (+/-)-platensimycin. AB - [reaction: see text] Reductive alkylation of 5-methoxy-1-tetralone (6) with 2,3 dibromopropene gave an equilibrium mixture of bicyclic diones 7 (51%) and 8 (35%). Radical cyclization of 7 afforded tricyclic dione 5 (84%), which was reduced, cyclized, and dehydrated to give tetracyclic alkene 13 in 63% yield. Allylic oxidation of 13 with SeO2 and activated MnO2 afforded enone 2 in 85% yield, thereby completing a short formal synthesis of (+/-)-platensimycin. PMID- 17407303 TI - Cross metathesis of alpha-methylene lactones II: gamma- and delta-lactones. AB - [reaction: see text] The cross metathesis reactivities of alpha-methylene-gamma butyrolactone and an alpha-methylene-delta-lactone have been investigated. alpha Methylene-gamma-butyrolactone undergoes rapid and efficient olefin isomerization in the presence of second-generation metathesis catalysts. However, cross metathesis can be achieved with the additive 2,6-dichlorobenzoquinone. In contrast, the alpha-methylene-delta-lactone neither isomerizes nor couples under similar conditions. PMID- 17407304 TI - Primary and secondary metabolite composition of kernels from three cultivars of Portuguese chestnut (Castanea sativa Mill.) at different stages of industrial transformation. AB - Chestnut (Castanea sativa) is an important basic food in rural diets and a major starch crop used in a similar way to potatoes. Chestnuts are a fundamental economic resource in the "chestnut regions" not only for the fruit but also for the chestnut wood. Chestnuts have become increasingly important with respect to human health, for example, as an alternative gluten-free flour source. Chestnuts are also a rich source of other beneficial compounds, but there have been few studies on the composition during processing. In this study, we analyzed the chemical composition of three Portuguese cultivars at different stages of industrial processing. The chestnut cultivars were Longal, Judia, and Martainha. All three cultivars had high moisture contents but were low in ash, crude fat, and crude protein contents, with high starch and low fiber contents. The free amino acid contents, including various essential amino acids, varied depending on the cultivar. All three cultivars also had a significant content of polyphenolics with gallic acid; ellagic acid was predominant among hydrolyzable and condensed tannins. Many of these compounds are known to exert significant positive effects on human health. The one-way analysis of variance for fresh chestnut shows significant differences among the three cultivars for most of the studied parameters. The same statistical analysis applied to each one of the two cultivars (Judia and Longal) sampled for the four processing steps analyzed indicates a significant effect of this factor in practically all of the constituents. On the other hand, the two-way analysis of variance shows that, besides the residual, the processing step and the interaction cultivar x processing step were the factors that more contributed for the total variation observed in the constituents analyzed, while the contribution of cultivar was much less significant. PMID- 17407305 TI - Toward metrological traceability for DNA fragment ratios in GM quantification. 1. Effect of DNA extraction methods on the quantitative determination of Bt176 corn by real-time PCR. AB - An international CCQM-P60 pilot study involving eight national metrological institutes was organized to investigate if the quantification of genetically modified (GM) corn powder by real-time PCR was affected by the DNA extraction method applied. Four commonly used extraction methods were compared for the extraction of DNA from a GM Bt176 corn powder. The CTAB-based method yielded the highest DNA template quantity and quality. A difference in the 260 nm/230 nm absorbance ratio was observed among the different extraction methods. Real-time amplification of sequences specific for endogenous genes zein and hmg as well as transgenic sequences within the cryIA(b) gene and a fragment covering the junction between the transformed DNA and the plant genome were used to determine the GM percentage. The detection of the transgenic gene was affected by the quantity and quality of template used for the PCR reaction. The Bt176 percentages measured on diluted or purified templates were statistically different depending on the extraction method applied. PMID- 17407306 TI - Toward metrological traceability for DNA fragment ratios in GM quantification. 2. Systematic study of parameters influencing the quantitative determination of MON 810 corn by real-time PCR. AB - This paper is part of a set of three papers investigating metrological traceability of the quantification of DNA fragments as, for instance, used for quantification of genetic modifications. This paper evaluates the possible impact of several factors on results of real-time Polymerase Chain Reaction (PCR) measurements. It was found that the particle size of the powder samples does not have an influence, whereas the nature of the calibrant (plasmidic or genomic DNA) has a significant effect. Moreover, two real-time PCR detection methods (construct-specific and event-specific) for MON 810 corn were compared. The results obtained in a specifically designed interlaboratory study revealed a significant influence of the DNA extraction method on measurement results when the MON 810 construct-specific real-time PCR detection method was applied. Statistical analyses confirmed the importance of validating DNA extraction methods in conjunction with real-time PCR methods. PMID- 17407307 TI - Toward metrological traceability for DNA fragment ratios in GM quantification. 3. Suitability of DNA calibrants studied with a MON 810 corn model. AB - The quantification of GMOs by real-time PCR relies on an external calibrant. In this paper the suitability of two DNA calibrants, genomic DNA from plant leaves and plasmidic DNA, was investigated. The PCR efficiencies, the correlation coefficients of the calibration curves, and the ratios between PCR efficiencies of transgenic and endogenous sequences were compared for both calibrants using 59 data sets produced by 43 laboratories. There were no significant differences between plasmidic and genomic DNA except for the PCR efficiencies of the calibration curves for the transgene of the construct-specific real-time PCR method. In the GM system investigated, PCR efficiencies of plasmidic calibrants were slightly closer to the PCR efficiencies observed for the unknowns than those of the genomic DNA calibrant. Therefore, plasmidic DNA was the more suitable calibrant for the PCR measurements on genomic DNA extracted from MON 810 seeds. It is shown that plasmidic DNA is an appropriate choice for the calibration of measurements of MON 810 corn with respect to the DNA copy number ratio. PMID- 17407308 TI - Synthesis of gluten-forming polypeptides. 1. Biosynthesis of gliadins and glutenin subunits. AB - Five winter wheat cultivars--GK Othalom (HMW-GS composition 2*, 7+8, 5+10), Ukrainka (1, 7+8, 5+10), Palotas (2*, 7+9, 5+10), Kodmon (2*, 7+8, 5+10), and Csongrad (2*, 7+9, 2+12)--grown in Hungary and harvested in the year 2005 were studied. The biosynthesis of gluten-forming polypeptides was followed starting at the 12th day after anthesis to the 53rd. Fresh kernel weight, moisture, and dry matter content of fresh kernels and gliadin and glutenin contents were determined. Gliadin components, total amounts of HMW and LMW polypeptides, and individual HMW polypeptides were determined using a RP-HPLC technique. Although considerable quantitative differences were observed concerning the content of total protein, gliadin, glutenin, and individual gluten-forming polypeptides, the character of accumulation of protein components--determined on the basis protein mass/kernel--was the same for the all of the cultivars studied and could be presented by a sigmoid curve. Small quantities of the gliadin and glutenin monomers may be detected in early stages of kernel development, but the bulk of these proteins is synthesized in later stages of development. It is generally suggested by specialists that the formation and accumulation of glutenin polymers starts later than the synthesis of monomers. Experimental data presented in this paper confirm this suggestion and show that in the first phase of protein synthesis the monomers are in "free" form; polymeric glutenin is detected only later. HMW glutenin subunits are synthesized synchronously, and quantitatively the polypeptides coded by chromosomes D and B dominate. PMID- 17407309 TI - Characterization of Vitis vinifera L. Cv. Carmenere grape and wine proanthocyanidins. AB - A formal compositional study of the proanthocyanidins of Vitis vinifera L. cv. Carmenere was conducted in this work. We first characterized the polymeric proanthocyanidins of Carmenere skins, seeds, and wines. In addition, the wine astringency was analyzed and compared with Cabernet Sauvignon. Although Carmenere wines had a higher proanthocyanidin concentration and mean degree of polymerization than Cabernet Sauvignon wines, the former wines were perceived as less astringent. The low seed/skin proportion in Carmenere wines as compared to other varieties, as evidenced by the reduced number of seeds per berry and the higher amount of epigallocatechin subunits of Carmenere wine proanthocyanidins, could explain this apparent paradox. PMID- 17407310 TI - Influence of soil type on the mobility and bioavailability of chelated zinc. AB - The objective of this study was to compare the distribution, mobility, and relative effectiveness of Zn from Zn-amino acids (Zn-AA) and Zn-DTPA-HEDTA-EDTA (Zn-CH) (DTPA, diethylenetriaminepentaacetate; HEDTA, N-2-hydroxyethyl ethylenedinitrilotriacetate; and EDTA, ethylenedinitrilotetraacetate) sources by applying different Zn levels to weakly acidic and neutral soils in laboratory (incubation and soil column studies) and greenhouse conditions. The experiments were carried out for 60 days in incubation and column experiments and for 45 days in a greenhouse experiment. The zinc soil behavior was evaluated by DTPA-TEA and Mehlich-3 extractions and sequential speciation. The incubation experiment showed that the highest concentrations of available Zn in weakly acidic soil occurred with Zn-AA treatments, whereas in the neutral soil Zn-CH treatments produced the highest quantities of available Zn. The column experiment showed that in neutral soil, with slow to moderate permeability in the Ap and Bt horizons, only Zn-CH significantly increased the mobility of Zn through the column with respect to the control and the Zn-AA source: 31% of the Zn applied as synthetic chelate was leached from the column. The greenhouse experiment showed that, at different rates of Zn application, the Zn carriers increased Zn uptake by maize (Zea mays L.). The use of applied Zn by maize, or Zn utilization, was greatest when the Zn treatments were Zn-CH (3.3%) at 20 mg kg-1 and Zn-CH (4.9%) at 10 mg kg-1, in weakly acidic and neutral soils, respectively. PMID- 17407311 TI - Breast cancer prevention by green tea catechins and black tea theaflavins in the C3(1) SV40 T,t antigen transgenic mouse model is accompanied by increased apoptosis and a decrease in oxidative DNA adducts. AB - Tea consumption is associated with a reduced risk of mammary cancer as reflected by epidemiological studies and experiments in carcinogen-induced rodent models of mammary carcinogenesis. We tested the hypothesis that green tea catechins (GTC) or theaflavins from black tea (BTT) interfere with mammary carcinogenesis in C3(1) SV40 T,t antigen transgenic multiple mammary adenocarcinoma (TAg) mice and that GTC/BTT affect tumor survival or oxidation status. TAg mice received GTC/BTT (0.05%) in drinking water for their lifetime. As compared to control mice, they survived longer and had smaller tumors. On microscopic inspection, the size of the largest tumor per mouse was decreased by 40-42% (p<0.01). GTC (0.01%) and BTT (0.05%) increased levels of cleaved caspase 3 in tumor tissue by 67 and 38%, respectively (p<0.05), intimating increased apoptosis. Tumor levels of the malondialdehyde-DNA adduct M1dG in mice receiving GTC or BTT (0.05%) were reduced by 78 (p<0.001) or 63% (p<0.05), respectively, as compared to controls. The results render the exploration of the breast cancer chemopreventive properties of tea preparations in humans worthwhile. PMID- 17407312 TI - Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit. AB - Polyphenol oxidase and peroxidase were extracted from two different varieties of strawberry fruit (Fragaria x ananassa D, cv. 'Elsanta' and Fragaria vesca L, cv. 'Madame Moutot') and characterized using reliable spectrophotometric methods. In all cases, the enzymes followed Michaelis-Menten kinetics, showing different values of peroxidase kinetics parameters between the two cultivars: Km = 50.68 +/ 2.42 mM ('Elsanta') and 18.18 +/- 8.79 mM ('Madame Moutot') mM and Vmax = 0.14 +/- 0.03 U/g ('Elsanta') and 0.05 +/- 0.01 U/g ('Madame Moutot'). The physiological pH of fruit at the red ripe stage negatively affected the expression of both oxidases, except polyphenol oxidase from 'Madame Moutot' that showed the highest residual activity (68% of the maximum). Peroxidase from both cultivars was much more thermolable as compared with PPO, losing over 60% of relative activity already after 60 min of incubation at 40 degrees C. The POD activation energy was much lower than the PPO activation energy (DeltaE = 97.5 and 57.8 kJ mol-1 for 'Elsanta' and 'Madame Moutot', respectively). Results obtained from d-glucose and d-fructose inhibition tests evidenced a decreasing course of PPO and POD activities from both cultivars as the sugar concentration in the assay medium increased. Changes in CIE L*, a*, b*, chroma, and hue angle values were taken as a browning index of the samples during storage at 4 degrees C. A decrease in L* was evident in both cultivars but more marked in 'Elsanta'. PPO and POD activities from cv. 'Elsanta' were very well-correlated with the parameter L* (r2=0.86 and 0.89, respectively) and hue angle (r2=0.85 and 0.93, respectively). According to these results, the browning of the fruit seemed to be in relation to both oxidase activities. PMID- 17407313 TI - Influence of thermal treatment applied to Fe(III) polyhydroxy cation intercalated vermiculite on the adsorption of atrazine. AB - Intercalation of vermiculite with Fe(III) polyhydroxy cations at 1:1 and 2:1 [OH ]/[Fe(III)] molar ratios increases the affinity of the clay mineral toward atrazine in comparison with potassium saturated vermiculite. The present paper describes the effects of thermal treatments applied to Fe(III) polyhydroxy cations modified vermiculite on the adsorption properties of the clay mineral. Only small changes in the textural characteristics were observed for the materials intercalated with either 1:1 or 2:1 [OH-]/[Fe(III)] molar ratios treated at 100 and 250 degrees C. In comparison with potassium saturated vermiculite, or intercalated vermiculite treated at 100 degrees C, a significant enhancement in the adsorption of atrazine was observed for the materials treated at 250 and 400 degrees C, which removed more than 95.8 and 99.5% of the herbicide initially present in a 50.0 microg L-1 aqueous solution, respectively. In comparison with potassium saturated vermiculite and intercalated vermiculite treated at 100 degrees C, a lower desorption degree of preadsorbed atrazine was observed for both intercalated materials treated at 250 and 400 degrees C. These findings suggest that the thermal treatment produced modified vermiculite materials with a high adsorption capacity and high affinity toward atrazine, with potential application in the removal of this herbicide, as well as other triazines, from aqueous medium. PMID- 17407314 TI - Determination of olive oil oxidative status by selected ion flow tube mass spectrometry. AB - The emergence of primary and secondary oxidation products in New Zealand extra virgin olive oil during accelerated thermal oxidation was measured and correlated with the concentrations of 13 headspace volatile compounds measured by selected ion flow tube mass spectrometry (SIFT-MS). SIFT-MS is a mass spectrometric technique that permits qualitative and absolute quantitative measurements to be made from whole air, headspace, or breath samples in real-time down to several parts per billion (ppb). It is well-suited to high-throughput analysis of headspace samples. Propanal, hexanal, and acetone were found at high concentrations in a rancid standard oil, while propanal, acetone, and acetic acid showed marked increases with oxidation time for the oils used in this study. A partial least-squares (PLS) regression model was constructed, which allowed the prediction of peroxide values (PV) for three separate oxidized oils. Sensory rancidity was also measured, although the correlations of headspace volatile compounds with sensory rancidity score were less satisfactory, and too few results were available for the construction of a PLS regression model. A fast (approximately 1 min), reliable method for prediction of olive oil PVs by SIFT-MS was developed. PMID- 17407315 TI - Variation in fatty acid composition of Artemia salina nauplii enriched with microalgae and baker's yeast for use in larviculture. AB - The high content of the essential fatty acids in some microalgae and baker's yeast has made them excellent diets for boosting the fatty acid content of livefood Artemia. The influences of baker's yeast (Saccharomyces cerevisiae) and three microalgae, viz., Chlorella salina, Chaetoceros calcitrans, and Nannochloropsis salina, were tested as diet components in marine livefeed brine shrimp Artemia salina nauplii to improve the polyunsaturated fatty acid (PUFA) composition. Artemia nauplii submerged in these diets for four different enrichment intervals (3, 6, 8, and 24 h) were found to incorporate essential fatty acids, and the percentage composition of different fatty acids was measured in the enriched Artemia nauplii and enrichment diets. N. salina produced higher levels of arachidonic acid (AA, 20:4n6, 9.50%), eicosapentaenoic acid (EPA, 20:5n3, 25.80%), and docosahexaenoic acid (DHA, 22:6n3, 4.18%) as compared to other diets. The total PUFA content of the enriched Artemia by N. salina increased by 56.50% with enrichment periods up to 8 h, followed by a significant reduction in the final 24 h. N. salina yielded Artemia nauplii with considerable EPA (8.05%), AA (14.15%), and DHA (1.85%) after 8 h of enrichment, which are significantly higher levels than in nauplii fed with the other three diets (p = 0.05). The DHA/EPA values in Artemia enriched for 6 h by N. salina and C. calcitrans were found to be, respectively, 88.46 and 25% higher than freshly hatched Artemia. Artemia enriched by C. salina and baker's yeast exhibited a reduction in PUFA content even at 6 h of enrichment. Significant relative decreases in DHA, EPA, and total PUFA in Artemia enriched with all of the diets were apparent, with a corresponding increase in the total saturated fatty acid content (26.95 +/- 9.75%) in the final stages (24 h) of enrichment (p = 0.05). PMID- 17407316 TI - Application of isotope dilution gas chromatography-mass spectrometry in analysis of organochlorine pesticide residues in ginseng root. AB - A highly accurate and precise method based on isotope dilution gas chromatography mass spectrometry was developed for the determination of five matrix-bound organochlorine pesticides, namely, hexachlorobenzene and hexachlorocyclohexanes (alpha-, beta-, delta-, and gamma- isomers), in a reference sample of Panax gingseng. Identification of the analytes was confirmed under selective ion monitoring mode by the presence of two dominant ion fragments within the specific time windows (+/-1% of the relative retention time with respect to the calibration standards) and matching of relative ion intensities of the concerned ions in samples and calibration standards (within +/-5%). Quantification was based on the measurement of concentration ratios of the natural and isotopic analogues in the sample and calibration blends. To circumvent the tedious iterative process of exact isotope matching that is often used in isotope dilution mass spectrometry analysis, a single-point calibration procedure was adopted with the isotopic amount ratios in the sample and calibration blends close to unity (0.9-1.1). Under the described approach, intraday and interday repeatability of replicate analyses of organochlorine pesticides in the ginseng root sample were below 1.4%. The expanded relative uncertainty ranging from 4.0 to 6.5% at a coverage factor of 2 was significantly lower than those of conventional gas chromatographic methods using other calibration techniques (internal or external standards). A deviation of less than 2.0% from the certified values was achieved when applying the developed method to determine hexachlorobenzene, alpha-, and beta-hexachlorocyclohexane in a certified reference material (CRM), BCR-CRM 115. Because of the unavailability of relevant CRMs of herbal origins, the concerned ginseng root sample, after verification of the "true values" of the concerned organochlorine pesticides by the valid primary method, is suitable for serving as an in-house reference material for quality assurance and method validation purposes. PMID- 17407317 TI - Lignin modification during Eucalyptus globulus kraft pulping followed by totally chlorine-free bleaching: a two-dimensional nuclear magnetic resonance, Fourier transform infrared, and pyrolysis-gas chromatography/mass spectrometry study. AB - Chemical modification of eucalypt lignin was investigated during kraft pulping and chlorine-free bleaching by comparing milled wood lignin, kraft lignin, and pulp enzymatic residual lignins. The syringyl-to-guaiacyl ratio (S/G) from analytical pyrolysis slightly changed during pulping and bleaching (S/G, 3-4) but was higher in the kraft lignin. Semiquantitative heteronuclear single quantum correlation (HSQC) nuclear magnetic resonance (NMR) showed that the relative amount of beta-O-4' (around 80% side chains) and resinol type substructures (15%) was slightly modified during pulping and oxygen delignification. However, a decrease of resinol substructures (to only 6%) was found after alkaline peroxide bleaching. The relative amount of surviving linkages in the highly phenolic kraft lignin was dramatically modified; resinols were predominant. Oxygen delignification did not change interunit linkages, but a relative increase of oxidized units was found in the HSQC aromatic region, in agreement with the small increase of pyrolysis markers with oxidized side chains. NMR heteronuclear multiple bond correlations showed that the oxidized units after oxygen delignification bore conjugated ketone groups. PMID- 17407318 TI - Effect of ionic detergents, nonionic detergents, and chaotropic agents on polyphenol oxidase activity from dormant saffron (Crocus sativus L.) corms. AB - Polyphenol oxidase (PPO; EC 1.14.18.1) catalyzes the hydroxylation of monophenols to o-diphenols (cresolase activity) and the oxidation of o-diphenols to o quinones (catecholase activity), leading to browning in plants and produce. Further interest in the enzyme has been triggered by the active role that it plays in plant defense systems. PPO can be found in latent forms and is activated in vitro by various agents including urea, detergents, and proteases. The activation of PPO from several sources by sodium dodecyl sulfate (SDS) has been extensively investigated, but reports on the effect of other detergents or on the differential effect of detergents on each of PPO's activities are scarce. In addition, investigations on the enzyme in other plant parts besides fruits and vegetables are also scarce. Here, the effect of various detergents and chaotropic agents on PPO from dormant saffron (Crocus sativus L.) corm extract was investigated. SDS and sarkosyl activated the cresolase activity, while only SDS activated the catecholase activity. All other detergents tested, in milli- or micromolar concentrations, inhibited the cresolase activity but barely affected the catecholase activity. In contrast, urea and guanidine-HCl drastically inhibited the catecholase activity but moderately inhibited the cresolase activity. The same effects were obtained on the partially purified enzyme. Results identified a PPO, present in dormant corms, which was activated only by anionic detergents and was inhibited by other reputed activating agents such as urea. Results also emphasized the differences in structure and accessibility of the active sites for cresolase and catecholase activities. PMID- 17407319 TI - Preparation of partially decaffeinated instant green tea. AB - The caffeine level of instant tea extracted from decaffeinated leaf tea with 4.0 mg g-1 caffeine is commonly above 10.0 mg g-1, the maximum limit of caffeine for decaffeinated instant tea. Further removal of caffeine by active carbon (AC) from the green tea extract was investigated. It showed that the removal of caffeine from the tea extract solutions depended on the treatment time and tea extract concentration while the ethanol concentration and pH had little effect on the removal of caffeine. According to the removal of caffeine and the ratio of total catechins to caffeine in the tested samples, the optimum decaffeination conditions were determined to be as follows: tea extract concentration 15-30 g L 1 for common tea extract but higher for partially decaffeinated tea leaf extract; ratio of tea solution to AC, 100 mL:4 g; treatment time, 4 h; and natural tea extract pH. Instant tea powder extracted from partially decaffeinated leaf tea with a caffeine level of 4.03 mg g-1 and further decaffeinated by AC had a caffeine level of 7.81 mg g-1, which was 31% lower than that without AC treatment. PMID- 17407320 TI - Norflurazon mobility, dissipation, activity, and persistence in a sandy soil as influenced by formulation. AB - Five ethylcellulose (EC) microencapsulated formulations (MEFs) of norflurazon were prepared and applied in soil to study their mobility, dissipation, activity, and persistence. The results show that the release into water of norflurazon from EC microspheres was retarded when compared with that of commercial herbicide. The mobility of norflurazon from MEFs into soil columns has been greatly diminished in comparison with that of its current commercial formulation (CF). Norflurazon distribution at different depths in the soil was higher in the upper ring (up to 50% of the initial application). In contrast, the residues from commercial norflurazon along the complete soil column were only about 2%. Degradation and bioassay experiments showed that the MEFs had greater persistence (t1/2 values were 7.72 and 30.83 weeks for CF and MEFs, respectively) and herbicidal activity than the commercial formulation. The use of these formulations can be advantageous, because they can minimize the risk of groundwater contamination and permit herbicide use at reduced rates, maintaining the desired concentrations of herbicide in the topsoil layer for longer periods of weed control. PMID- 17407321 TI - Differentiation of young red wines based on chemometrics of minor polyphenolic constituents. AB - For reasons of examining any possible discrimination of young red wines based on selected, minor polyphenols, a survey was carried out including 35 samples originating from three Hellenic native and three international Vitis vinifera cultivars from various regions of Greece. All samples were experimental wines vinified and stored under identical conditions, in an effort to minimize the effect of different winemaking technologies as well as aging. The polyphenols analyzed belonged to two categories: benzoic acid derivatives, including gallic acid, protocatechuic acid, vanillic acid, and syringic acid, and stilbenes, including astringin, piceid, and resveratrol (all trans isomers). Data handling employing discriminant analysis (DA) yielded very satisfactory categorization of samples in terms of both cultivar and geographical region of origin. This outcome was discussed with regard to the value of certain minor polyphenols that could serve as characteristic indices for discrimination of varietal red wines, after appropriate implementation of chemometrics. PMID- 17407322 TI - D-amino acid formation in sterilized alkali-treated olives. AB - The occurrence of d-amino acids in commercial ripe olives, a well-known sterilized alkali-treated product, was investigated by high-performance liquid chromatography (HPLC) with precolumn automatic derivatization. Absolute amounts of D-amino acids were in total 18.6-38.2 mg/100 g edible portion. The major D amino acids were D-aspartic acid, D-glutamic acid, D-serine, and D-leucine. Furthermore, to evaluate the effects of sterilization time and olive pH on amino acid racemization, a simulated processing of green ripe olives was carried out. Serine (both free and bound form) was the most-racemized amino acid after heat treatment. Sterilization (15-35 min at 121 degrees C) increased the racemization values of both free and protein-bound amino acids, although in case of protein bound phenylalanine the increase was not statistically significant. With an increase of pH from 8 to 10 units, the racemization values of all amino acids increased significantly, except for free forms of aspartic and glutamic acids. In general, the effects of the sterilization time and olive pH on total concentration (L + D enantiomers) of each amino acid were also significant. PMID- 17407323 TI - Preparation and characterization of inclusion complex of iprodione and beta cyclodextrin to improve fungicidal activity. AB - The effect of beta-cyclodextrin (beta-CD) on the improvement of the fungicidal activity of iprodione has been investigated. The inclusion complexation of beta CD with iprodione has been prepared and characterized by integrating some analytical techniques (such as electrospray ionization-mass spectrometry, differential scanning calorimetry, thermogravimetry, x-ray diffraction, and scanning electron microscopy) and molecular simulation methods. The beta CD/iprodione inclusion complex has exhibited different spectroscopic features and properties from iprodione. The stoichiometric ratio and stability constant describing the extent of formation of inclusion complexes have been determined by phase solubility studies. The calculated apparent stability constant of the iprodione/beta-CD complex was 407.5 M-1. The obtained inclusion complexes were found to significantly improve the water solubility of iprodione, and there is a 4.7-fold increase in the presence of 13 mM beta-CD as compared with the solubility of iprodione in deionized water in the absence of beta-CD. The bioassay demonstrated that the complex displayed over two-fold increase of the fungicidal activity. In addition, the possible structure of the beta-CD/iprodione complex was proposed according to the results of the molecular dynamic simulation. The present study provided useful information for a more rational application of iprodione, diminishing the use of organic solvents and increasing its efficacy. PMID- 17407324 TI - MiaB, a bifunctional radical-S-adenosylmethionine enzyme involved in the thiolation and methylation of tRNA, contains two essential [4Fe-4S] clusters. AB - The radical-S-adenosylmethionine (radical-AdoMet) enzyme MiaB catalyzes the posttranscriptional methylthiolation of N-6-isopentenyladenosine in tRNAs. Spectroscopic and analytical studies of the reconstituted wild-type and C150/154/157A triple variant forms of Thermotoga maritima MiaB have revealed the presence of two distinct [4Fe-4S](2+,1+) clusters in the protein. One is coordinated by the three conserved cysteines in the radical-AdoMet motif (Cys150, Cys154, and Cys157) as previously reported, and the other, here observed for the first time, is proposed to be coordinated by the three N-terminal conserved cysteines (Cys10, Cys46, and Cys79). The two [4Fe-4S]2+ clusters have similar UV visible absorption, resonance Raman, and Mossbauer properties but differ in terms of redox properties and the EPR properties of the reduced [4Fe-4S]1+ clusters. Reconstituted forms of MiaB containing two [4Fe-4S] clusters are more active than previously reported. Comparison of MiaB with other radical-AdoMet enzymes involved in thiolation reactions, such as biotin synthase and lipoate synthase, is discussed as well as a possible role of the second cluster as a sacrificial S donor in the MiaB-catalyzed reaction. PMID- 17407325 TI - Neighboring group participation in the transition state of human purine nucleoside phosphorylase. AB - The X-ray crystal structures of human purine nucleoside phosphorylase (PNP) with bound inosine or transition-state analogues show His257 within hydrogen bonding distance of the 5'-hydroxyl. The mutants His257Phe, His257Gly, and His257Asp exhibited greatly decreased affinity for Immucillin-H (ImmH), binding this mimic of an early transition state as much as 370-fold (Km/Ki) less tightly than native PNP. In contrast, these mutants bound DADMe-ImmH, a mimic of a late transition state, nearly as well as the native enzyme. These results indicate that His257 serves an important role in the early stages of transition-state formation. Whereas mutation of His257 resulted in little variation in the PNP x DADMe-ImmH x SO4 structures, His257Phe x ImmH x PO4 showed distortion at the 5'-hydroxyl, indicating the importance of H-bonding in positioning this group during progression to the transition state. Binding isotope effect (BIE) and kinetic isotope effect (KIE) studies of the remote 5'-(3)H for the arsenolysis of inosine with native PNP revealed a BIE of 1.5% and an unexpectedly large intrinsic KIE of 4.6%. This result is interpreted as a moderate electronic distortion toward the transition state in the Michaelis complex with continued development of a similar distortion at the transition state. The mutants His257Phe, His257Gly, and His257Asp altered the 5'-(3)H intrinsic KIE to -3, -14, and 7%, respectively, while the BIEs contributed 2, 2, and -2%, respectively. These surprising results establish that forces in the Michaelis complex, reported by the BIEs, can be reversed or enhanced at the transition state. PMID- 17407326 TI - Conformation of an endogenous ligand in a membrane bilayer for the macrophage scavenger receptor CD36. AB - Phagocytic removal of aged or oxidatively damaged cells and macromolecules is an indispensable homeostatic function of the innate immune system. A structurally conserved family of oxidized phospholipids that serve as endogenous high-affinity ligands for the macrophage scavenger receptor CD36 (oxPC(CD36)) was recently identified. Enriched within atherosclerotic plaque and senescent cell membranes, oxPC(CD36) promote the uptake of oxidized lipoproteins and cell membranes by macrophages when present at only a few molecules per particle. How macrophages recognize oxPC(CD36) within cellular membranes and lipoprotein surfaces remains unknown. Herein, we deduce the conformation of oxPC(CD36) near the hydrophobic hydrophilic interface within membrane bilayers by determining multiple critical internuclear distances using nuclear Overhauser enhancement spectroscopy. The molecular model reveals a unique conformation for oxPC(CD36) within bilayers whereby the distal end of the sn-2 acyl chain harboring the structurally conserved CD36 recognition motif protrudes into the aqueous phase. The remarkable conformation elucidated for oxPC(CD36) produces a surface accessible phagocytic "eat me signal" to facilitate senescent cell and oxidized lipoprotein recognition by the scavenger receptor CD36 as part of its immune surveillance function. PMID- 17407327 TI - Crystal structures of human carboxylesterase 1 in covalent complexes with the chemical warfare agents soman and tabun. AB - The organophosphorus nerve agents sarin, soman, tabun, and VX exert their toxic effects by inhibiting the action of human acetylcholinesterase, a member of the serine hydrolase superfamily of enzymes. The current treatments for nerve agent exposure must be administered quickly to be effective, and they often do not eliminate long-term toxic side effects associated with organophosphate poisoning. Thus, there is significant need for effective prophylactic methods to protect at risk personnel from nerve agent exposure, and protein-based approaches have emerged as promising candidates. We present the 2.7 A resolution crystal structures of the serine hydrolase human carboxylesterase 1 (hCE1), a broad spectrum drug metabolism enzyme, in covalent acyl-enzyme intermediate complexes with the chemical weapons soman and tabun. The structures reveal that hCE1 binds stereoselectively to these nerve agents; for example, hCE1 appears to react preferentially with the 10(4)-fold more lethal PS stereoisomer of soman relative to the PR form. In addition, structural features of the hCE1 active site indicate that the enzyme may be resistant to dead-end organophosphate aging reactions that permanently inactivate other serine hydrolases. Taken together, these data provide important structural details toward the goal of engineering hCE1 into an organophosphate hydrolase and protein-based therapeutic for nerve agent exposure. PMID- 17407328 TI - Covalent adducts arising from the decomposition products of lipid hydroperoxides in the presence of cytochrome c. AB - Polyunsaturated fatty acids can be converted to lipid hydroperoxides through nonenzymatic and enzymatic pathways. The prototypic omega-6 lipid hydroperoxide 13-hydroperoxy-octadecadienoic acid (13-HPODE) homolytically decomposes to form highly reactive alpha,beta-unsaturated aldehydes, such as 9,12-dioxo-10(E) dodecenoic acid (DODE), 4-oxo-2(E)-nonenal (ONE), 4,5-epoxy-2(E)-decenal (EDE), and 4-hydroxy-2(E)-nonenal (HNE), that can form covalent adducts with DNA. Both 4 oxo-2(E)-nonenal and 4-hydroxy-2(E)-nonenal can also modify proteins to form products that can potentially serve as biomarkers of lipid hydroperoxide-mediated macromolecule damage. In this study, cytochrome c was used to identify and individually characterize the modification sites for each of these aldehydes and also determine the most abundant adduct formed following the decomposition of 13 HPODE. The adducts were characterized by ESI-TOF/MS analysis of the intact proteins and by a combination of ESI-ion-trap/MSn and quadrupole-TOF/MS/MS analysis of the tryptic and chymotryptic peptides. The major adducts included an HNE-His Michael adduct on H33, EDE-Lys adducts on K7 and K8, ONE-Lys ketoamide adducts on K5, K7, and K8, an apparent ONE-Lys Michael adduct on K5, and DODE-Lys carboxyl ketoamide adducts on K86 and K87. DODE was the most reactive aldehyde toward cytochrome c. The major adduct from this reaction was analogous to the most abundant adduct resulting from the decomposition of 13-HPODE in the presence of cytochrome c. PMID- 17407329 TI - Human cytochrome p450 enzyme specificity for the bioactivation of estragole and related alkenylbenzenes. AB - Human cytochrome P450 enzymes involved in the bioactivation of estragole to its proximate carcinogen 1'-hydroxyestragole were identified and compared to the enzymes of importance for 1'-hydroxylation of the related alkenylbenzenes methyleugenol and safrole. Incubations with Supersomes revealed that all enzymes tested, except P450 2C8, are intrinsically able to 1'-hydroxylate estragole. Experiments with Gentest microsomes, expressing P450 enzymes to roughly average liver levels, indicated that P450 1A2, 2A6, 2C19, 2D6, and 2E1 might contribute to estragole 1'-hydroxylation in the human liver. Especially P450 1A2 is an important enzyme based on the correlation between P450 1A2 activity and estragole 1'-hydroxylation in human liver microsomal samples and inhibition of estragole 1' hydroxylation by the P450 1A2 inhibitor alpha-naphthoflavone. Kinetic studies revealed that, at physiologically relevant concentrations of estragole, P450 1A2 and 2A6 are the most important enzymes for bioactivation in the human liver showing enzyme efficiencies (kcat/Km) of, respectively, 59 and 341 min-1 mM-1. Only at relatively high estragole concentrations, P450 2C19, 2D6, and 2E1 might contribute to some extent. Comparison to results from similar studies for safrole and methyleugenol revealed that competitive interactions between estragole and methyleugenol 1'-hydroxylation and between estragole and safrole 1'-hydroxylation are to be expected because of the involvement of, respectively, P450 1A2 and P450 2A6 in the bioactivation of these compounds. Furthermore, poor metabolizer phenotypes in P450 2A6 might diminish the chances on bioactivation of estragole and safrole, whereas lifestyle factors increasing P450 1A2 activities such as cigarette smoking and consumption of charbroiled food might increase those chances for estragole and methyleugenol. PMID- 17407330 TI - Atomic-scale friction on diamond: a comparison of different sliding directions on (001) and (111) surfaces using MD and AFM. AB - Atomic force microscopy (AFM) experiments and molecular dynamics (MD) simulations were conducted to examine single-asperity friction as a function of load, surface orientation, and sliding direction on individual crystalline grains of diamond in the wearless regime. Experimental and simulation conditions were designed to correspond as closely as state-of-the-art techniques allow. Both hydrogen terminated diamond (111)(1 x 1)-H and the dimer row-reconstructed diamond (001)(2 x 1)-H surfaces were examined. The MD simulations used H-terminated diamond tips with both flat- and curved-end geometries, and the AFM experiments used two spherical, hydrogenated amorphous carbon tips. The AFM measurements showed higher adhesion and friction forces for (001) vs (111) surfaces. However, the increased friction forces can be entirely attributed to increased contact area induced by higher adhesion. Thus, no difference in the intrinsic resistance to friction (i.e., in the interfacial shear strength) is observed. Similarly, the MD results show no significant difference in friction between the two diamond surfaces, except for the specific case of sliding at high pressures along the dimer row direction on the (001) surface. The origin of this effect is discussed. The experimentally observed dependence of friction on load fits closely with the continuum Maugis-Dugdale model for contact area, consistent with the occurrence of single-asperity interfacial friction (friction proportional to contact area with a constant shear strength). In contrast, the simulations showed a nearly linear dependence of the friction on load. This difference may arise from the limits of applicability of continuum mechanics at small scales, because the contact areas in the MD simulations are significantly smaller than the AFM experiments. Regardless of scale, both the AFM and MD results show that nanoscale tribological behavior deviates dramatically from the established macroscopic behavior of diamond, which is highly dependent on orientation. PMID- 17407331 TI - Thermal restructuring of fractal clusters: the case of a strawberry-like core shell polymer colloid. AB - Thermal restructuring of fractal styrene-acrylate copolymer clusters dispersed in water has been investigated experimentally in the temperature range between 313 and 363 K. The particles constituting the clusters are of strawberry-like core shell structure with a soft core and a rigid shell grafted on the core polymer chains. Due to the incomplete coverage of the core, the rather soft core may "flow out" through the open areas of the shell, leading to coalescence with the neighboring particles. The clusters were generated under diffusion-limited cluster aggregation conditions, and the restructuring kinetics was monitored by small-angle light scattering. Two sets of thermal restructuring experiments have been performed at various temperatures: (1) restructuring of growing clusters during aggregation and (2) restructuring of preformed clusters in the absence of aggregation. It is found that restructuring occurs only at temperature values above 323 K. In the absence of aggregation, restructuring leads to an increase of the fractal dimension and a decrease of the radius of gyration of the clusters. At sufficiently long times, both quantities reach a plateau value due to the presence of the grafted rigid shell, which constrains the coalescence of the soft core. A simple model, based on coalescence theory of liquid droplets and accounting for the incomplete coalescence and its dependence on temperature, has been developed to interpret the restructuring kinetics in the absence of aggregation. It is found that the proposed model can represent the measured experimental data well. PMID- 17407332 TI - Modeling the structure formation of particulate langmuir films: the effect of polydispersity. AB - Two-dimensional molecular dynamics computer simulation has been developed to model the compression of Langmuir films composed of spherical nanoparticles with arbitrary size distribution. We demonstrate that the usual assumption in the determination of interparticle potentials from the surface pressure vs area isotherms (i.e., monodisperse particles in perfect hexagonal order) leads to a systematic overestimation of the characteristic length of the interaction. On the basis of the results of the simulation, we propose a correction method to improve the traditional way of determining the interparticle potentials. We use the corrected particle-particle interactions to explore the correlation between the broadness of the size distribution and several structural parameters (decay length of pair-correlation function, global orientational order parameter, mean, and standard deviation of number of neighbors). Due to the uniaxial compression and the stiffness of the particulate layer, the surface pressure is not a scalar field. We investigate the effect of polydispersity on the anisotropy and the fluctuation of the surface pressure tensor in Langmuir films during uniaxial compression. PMID- 17407333 TI - Synthesis of size-controlled and shaped copper nanoparticles. AB - The synthesis of stable, monodisperse, shaped copper nanoparticles has been difficult, partially because of copper's propensity for oxidation. This article reports the findings of an investigation of a synthetic route for the synthesis of size-controllable and potentially shape-controllable molecularly capped copper nanoparticles. The approach involved the manipulation of reaction temperature for the synthesis of copper nanoparticles in organic solvents in the presence of amine and acid capping agents. By manipulating the reaction temperature, this route has been demonstrated for the production of copper nanoparticles ranging from 5 to 25 nm. The size dependence of the melting temperature of copper nanoparticles, especially for surface melting, is believed to play an important role in interparticle coalescence, leading to size growth as the reaction temperature is increased. Control of the reaction temperature and capping molecules has also been demonstrated to produce copper nanoparticles with different shapes such as rods and cubes. The previously proposed combination of the selective formation of a seed precursor and a selective growth direction due to the preferential adsorption of capping agents on certain nanocrystal facets is believed to be responsible for shape formation by kinetically controlling the growth rates of crystal facets. The nanoparticles are characterized using TEM, XRD, and UV-visible techniques. A mechanistic consideration of the size control and shape formation is also discussed. PMID- 17407334 TI - Fluorescence from an azobenzene-containing diblock copolymer micelle in solution. AB - We report the observation of unusual fluorescence emission from an azobenzene containing polymer micellar solution. An amphiphilic diblock copolymer composed of the hydrophilic quaternized poly(4-vinyl pyridine) (QP4VP) and a hydrophobic liquid crystalline polymethacrylate bearing azobenzene side groups (PAzoMA) is nonfluorescent in molecularly dissolved state in N,N-dimethyl formamide (DMF) but becomes fluorescent as a result of the micellization upon addition of water, which confines azobenzene groups into the core region of micellar aggregates. Experimental results suggest that the micellization-enhanced fluorescence was caused by a slowdown, due to the confinement effect, in the rate of the trans-to cis photoisomerization that is the main nonradiative relaxation process for excited azobenzene groups in the trans form. Furthermore, it was found that the fluorescence intensity of aqueous micellar solution is sensitive to changes in pH (reversible fluorescence variation) and to illumination (irreversible fluorescence variation). The results indicate that a subtle change in the state of polymer micellar association may alter the confining state of azobenzene groups responsible for the fluorescence emission. PMID- 17407335 TI - Nanotemplating for two-dimensional molecular imprinting. AB - A new 2D molecular imprinting technique based on nanotemplating and soft lithography techniques is reported. This technique allows the creation of target specific synthetic recognition sites on different substrates using a uniquely oriented and immobilized template and the attachment of a molecularly imprinted polymer on a substrate. The molecularly imprinted polymer was characterized by AFM, fluorescence microscopy, and ATR-FTIR. We evaluated the rebinding ability of the sites with theophylline (the target molecule). The selectivity of the molecularly imprinted polymer was determined for the theophylline-caffeine couple. The molecularly imprinted polymer exhibited selectivity for theophylline, as revealed by competitive rebinding experiments. Fluorescence microscopy experiments provided complementary proof of the selectivity of the molecularly imprinted polymer surfaces toward theophylline. These selective molecularly imprinted polymers have the potential for chemical sensor applications. Because of its 2D nature, this novel chemical sensor technology can be integrated with many existing high-sensitivity multichannel detection technologies. PMID- 17407336 TI - Proton activity in nafion films: probing exchangeable protons with methylene blue. AB - A simple approach to monitor the H(+) activity of a proton-exchange membrane (Nafion) is introduced by incorporating methylene blue as an indicator dye. The dye exhibits characteristics absorption maxima at 665 and 745 nm corresponding to its singly and doubly protonated forms, respectively. The apparent proton activity of Nafion as monitored from the appearance of doubly protonated methylene blue absorption is equivalent to 1.2 M H2SO4. By monitoring the spectral changes associated with the protonation equilibrium of the dye, it is possible to probe the rate and the exchangeable proton sites within the Nafion film. For the Nafion 117 film, we estimate the total exchangeable proton sites to be 2.5 x 10(19) sites/cm(2) or 4.2 x 10(-5) mols/cm(2). The equilibrium constant for the H(+)/Na(+) exchange for the bound sites is determined to be 2.2. The feasibility of methylene blue as a probe to monitor proton activity during the operation of a direct methanol fuel cell has been explored. PMID- 17407337 TI - Cell adhesion properties on photochemically functionalized diamond. AB - The biocompatibility of diamond was investigated with a view toward correlating surface chemistry and topography with cellular adhesion and growth. The adhesion properties of normal human dermal fibroblast (NHDF) cells on microcrystalline and ultrananocrystalline diamond (UNCD) surfaces were measured using atomic force microscopy. Cell adhesion forces increased by several times on the hydrogenated diamond surfaces after UV irradiation of the surfaces in air or after functionalization with undecylenic acid. A direct correlation between initial cell adhesion forces and the subsequent cell growth was observed. Cell adhesion forces were observed to be strongest on UV-treated UNCD, and cell growth experiments showed that UNCD was intrinsically more biocompatible than microcrystalline diamond surfaces. The surface carboxylic acid groups on the functionalized diamond surface provide tethering sites for laminin to support the growth of neuron cells. Finally, using capillary injection, a surface gradient of polyethylene glycol could be assembled on top of the diamond surface for the construction of a cell gradient. PMID- 17407338 TI - Study of confined 5-aza[5]helicene in ytterbium(III) bis(2-ethylhexyl) sulfosuccinate reversed micelles. AB - Some relevant physicochemical properties of 5-aza[5]helicene (H5) in solutions of ytterbium bis(2-ethylhexyl) sulfosuccinate (Yb(DEHSS)3) reversed micelles have been investigated by UV-vis-NIR, photoluminescence, and FT-IR techniques with the aim of emphasizing the role played by specific Yb(III)/H5 interactions and confinement effects as driving forces of its binding to reversed micelles, preferential solubilization site, and local photophysical properties. It has been found that the binding strength of 5-aza[5]helicene to reversed micelles, triggered by steric and orientational constrains as well as the water content, is mainly regulated by its interaction with the Yb(III) counterion. Moreover, when H5 is entrapped in Yb(DEHSS)3 reversed micelles, the combined action of this interaction and of confinement effects leads to marked changes of its photophysical properties with respect to those of H5 molecularly dispersed in apolar medium. The influence of the entrapment of finite amounts of H5 on the reversed micelle structure was investigated by SAXS. The analysis of experimental results brings to the hypothesis that H5 is preferentially solubilized and opportunely oriented in the micellar palisade layer and that its insertion causes an unidimensional growth of reversed micelles. From an analysis of WAXS spectra of H5/Yb(DEHSS)3 composites, obtained by complete evaporation of the volatile components of the H5/water/Yb(DEHSS)3/n-heptane solutions, it was ascertained that also on these systems H5 is dispersed molecularly or in a quite amorphous state in the surfactant liquid crystals without forming a separate crystalline nanophase. PMID- 17407339 TI - L-alanine in a droplet of water: a density-functional molecular dynamics study. AB - We report the results of a Born-Oppenheimer molecular dynamics study on an L alanine amino acid in neutral aqueous solution. The whole system, the L-alanine zwitterion and 50 water molecules, was treated quantum mechanically. We found that the hydrophobic side chain (R = CH3) defines the trajectory path of the molecule. Initially fully hydrated in an isolated droplet of water, the amino acid moves to the droplet's surface, exposing its hydrophobic methyl group and alpha-hydrogen out of the water. The structure of an L-alanine with the methyl group exposed to the water surface was found to be energetically favorable compared to a fully hydrated molecule. The dynamic behavior of the system suggests that the first hydration shell of the amino acid is localized around carboxylate (CO2-) and ammonium (NH3+) functional groups; it is highly ordered and quite rigid. In contrast, the hydration shell around the side chain is much less structured, suggesting a modest influence of the methyl group on the structure of water. The number of water molecules in the first hydration shell of an alanine molecule is constantly changing; the average number was found to equal 7. The molecular dynamics results show that L-alanine in water does not have a preferred conformation, as all three of the molecule's functional sites (i.e., CH3, NH3+, CO2-) perform rotational movements around the C(alpha)-site bond. PMID- 17407340 TI - Mossbauer characterization and in situ monitoring of thermal decomposition of potassium ferrate(VI), K2FeO4 in static air conditions. AB - Solid orthorhombic crystals of potassium ferrate(VI) (K(2)FeO(4)) of a high chemical purity (>99.0%) were characterized by low-temperature (1.5-5 K), high temperature (463-863 K), and in-field (1.5 K/3 T) Mossbauer spectroscopy. Potassium ferrate(VI) reveals a Neel magnetic transition temperature (TN) of approximately 3.8 K and a saturation hyperfine magnetic field of 13.8 T at 1.5 K. Spectral line intensities recorded below TN in an external magnetic field of 3 T manifest a perfect antiferromagnetic ordering. For the in situ monitoring of the thermal behavior of K(2)FeO(4), high-temperature Mossbauer data were combined with those obtained from thermogravimetry, differential scanning calorimetry, and variable-temperature X-ray diffraction measurements. Such in situ approach allowed the identification of the reaction products and intermediates and yielded the first experimental evidence for the participation of CO2 in the decomposition process. As the primary conversion products, KFeO(2) and two potassium oxides in equivalent molar ratio, KO2 and K(2)O, were suggested. However, the KO2 phase is detectable with difficulty as it reacts very quickly with CO2 from air resulting in the formation of K(2)CO(3). The presented decomposition model is consistent with thermogravimetric data giving the mass loss of 8.0%, which corresponds to the participation of 1/6 mol of CO2 and liberation of 3/4 mol of O2 per 1 mol of K(2)FeO(4) (K(2)FeO(4) + 1/6CO2 --> KFeO(2) + 1/3K(2)O + 1/6K(2)CO(3) + 3/4O2). An explanation of the multistage reaction mechanism has an important practical impact for the optimization of the solid-state synthesis of potassium ferrate(VI). PMID- 17407341 TI - Immobilization of hexokinase onto chitosan decorated particles. AB - Chitosan (CH) decorated polystyrene (PS) particles were synthesized within complexes of CH, a polycation under acid conditions, and tiny amounts of sodium dodecylsulfate (SDS). Particle characterization was performed by means of dynamic light scattering, zeta potential measurements, and transmission electron microscopy. All dispersions were stable in the ionic strength of 2.0 mol L-1 NaCl during 2 months. The outstanding colloidal stability was attributed to the presence of a hydrated CH layer around the particles. CH decorated PS particles were attached to atomic force microscopy cantilevers and probed against Si wafers in water and in NaCl 0.01 mol/L. The mean thickness of CH layer amounted to 35 +/ 11 and 16 +/- 6 nm, when the medium was water and NaCl 0.01 mol/L, respectively. Adsorption isotherm of hexokinase (HK) onto PS/CH particles studied by means of spectrophotometry showed three regions: an initial step; adsorption plateau and multilayer formation. Enzymatic activity of free HK and immobilized HK was monitored by means of spectrophotometry as a function of storing time and reuse. After 3 days, storing HK free in solution dramatically lost its catalytic properties. On the contrary, HK-covered PS/CH particles retained enzymatic activity over 1 month. Moreover, HK-covered PS/CH particles could be reused in the determination of glucose two times consecutively, without losing activity. These interesting findings were discussed in light of the role of water in enzyme conformation. PMID- 17407342 TI - Multidimensional potentials of mean force from biased experiments along a single coordinate. AB - External biasing forces are often applied to enhance sampling in regions of phase space which would otherwise be rarely observed. While the typical goal of these experiments is to calculate the potential of mean force (PMF) along the biasing coordinate, here I present a method to construct PMFs in multiple dimensions and along arbitary alternative degrees of freedom. A protocol for multidimensional PMF reconstruction from nonequilibrium single-molecule pulling experiments is introduced and tested on a series of two-dimensional potential surfaces with varying levels of correlation. Reconstruction accuracy and convergence from several methods--this new protocol, equilibrium umbrella sampling, and free diffusion--are compared, and nonequilibrium pulling is found to be the most efficient. To facilitate the use of this method, the source code for this analysis is made freely available. PMID- 17407343 TI - Direct evidence of layer-by-layer assembly of polyelectrolyte multilayers on soft and porous temperature-sensitive PNiPAM microgel using fluorescence correlation spectroscopy. AB - We describe the layer-by-layer assembly of polyelectrolyte multilayers on soft and porous temperature-sensitive poly(N-isopropylacrylamide) (PNiPAM) microgel. Microgels are not hard and rigid but rather are soft and porous particles, and polyelectrolytes not only interdigitate with each other during multilayer formation but also with the microgel. Because of this difference, there could be concerns about the feasibility of the layer-by-layer technique on these systems. The argument is that the layer being deposited is stripping the underlying layer instead of anchoring to the latter, and common methods of characterizing film growth on particles such as zeta-potentials will still show "successful" charge reversal. To address this issue, we used two differently labeled polyelectrolytes during the deposition. Because of the small size of the microgel (400 nm) studied, we cannot distinguish between polyelectrolytes adsorbed on or in the microgel. However, with fluorescence correlation spectroscopy, we can clearly distinguish between free labeled polyelectrolytes and those that are bound to the microgel. Dual-color correlation confirms the presence of both polyelectrolytes bound to the same particle while fluorescence imaging (on a dry sample) provides the visual proof. PMID- 17407344 TI - Bimodal swelling responses in microgel thin films. AB - A series of studies on microgel thin films is described, wherein quartz crystal microgravimetry (QCM), surface plasmon resonance (SPR), and atomic force microscopy (AFM) have been used to probe the properties of microstructured polymer thin films as a function of film architecture and solution pH. Thin films composed of pNIPAm-co-AAc microgels were constructed by using spin-coating layer by-layer (scLbL) assembly with poly(allylamine hydrochloride) (PAH) as a polycationic "glue". Our findings suggest that the interaction between the negatively charged microgels and the positively charged PAH has a significant impact on the pH responsivity of the film. These effects are observable in both the optical and mechanical behaviors of the films. The most significant changes in behavior are observed when the motional resistance of a quartz oscillator is monitored via QCM experiments. Slight changes to the film architecture and alternating the pH of the environment significantly changes the QCM and SPR responses, suggesting a pH-dependent swelling that is dependent on both particle swelling and polyelectrolyte de-complexation. Together, these studies allow for a deeper understanding of the morphological changes that take place in environmentally responsive microgel-based thin films. PMID- 17407345 TI - High-pressure studies of cyclohexane to 40 GPa. AB - We present data from two room temperature synchrotron X-ray powder diffraction studies of cyclohexane up to approximately 40 and approximately 20 GPa. In the first experiment, pressure cycling was employed wherein pressure was varied up to approximately 16 GPa, reduced to 3.5 GPa, and then raised again to 40 GPa. Initially, the sample was found to be in the monoclinic phase (P12(1)/n1) at approximately 8.4 GPa. Beyond this pressure, the sample adopted triclinic unit cell symmetry (P1) which remained so even when the pressure was reduced to 3.5 GPa, indicating significant hysteresis and metastability. In the second experiment, pressure was more slowly varied, and the monoclinic unit cell structure (P12(1)/n1) was observed at lower pressures up to approximately 7 GPa, above which a phase transformation into the P1 triclinic unit cell symmetry occurred. Thus, the pressure onset of the triclinic phase may be dependent upon the pressurizing conditions. High-pressure Raman data that further emphasize a phase transition (probably into phase VI) around 10 GPa are also presented. We also have further evidence for a phase VII, which is probably triclinic. PMID- 17407346 TI - Kinetics of the electron self-exchange and electron-transfer reactions of the (trimethylammonio)methylferrocene host-guest complex with cucurbit[7]uril in aqueous solution. AB - The electron self-exchange rate constants for the (trimethylammonio)methylferrocene(+/2+) couple (FcTMA+/2+) have been measured in the absence and presence of the cucurbit[7]uril (CB[7]) host molecule in aqueous solution, using 1H NMR line-broadening experiments. The very strong binding of the ferrocene to CB[7] results in slow exchange of the guest on the NMR time scale, such that resonances for both the free and bound forms of the reduced ferrocene can be observed. The extents of line broadening in the resonances of the two forms of the guest in the presence of the FcTMA2+ species can be monitored independently, allowing for the determination of the rate constants for the possible self-exchange pathways involving the bound and free forms of both the oxidized and reduced members of the redox couple. The encapsulation of both the reduced and oxidized forms of the ferrocene increases the rate constant (25 degrees C) from (2.1+/-0.1)x10(6) M-1 s-1 (for FcTMA+/2+) to (6.7+/-0.7)x10(6) M( 1) s(-1) (for {FcTMA.CB[7]}+/2+), whereas inclusion of the reduced form only decreases the rate constant to (6+/-1)x10(5) M(-1) s(-1). The changes in the exchange rate constants upon inclusion of the reactants are related to the effects of CB[7] acting as an outer, second-coordination sphere and are compared to those observed previously for the electron-exchange process in the presence of beta-cyclodextrin and p-sulfonated calix[6]arene hosts. The binding of FcTMA+ and hydroxymethylferrocene to CB[7] significantly reduces the rate constants for their oxidations by the bis(2,6-pyridinedicarboxylato)cobaltate(III) ion (which does not bind to CB[7]) as a result of reduced thermodynamic driving forces and steric hindrance to close approach of the oxidant to the encapsulated ferrocenes. PMID- 17407347 TI - Side-chain and backbone ordering in homopolymers. AB - In order to study the relation between backbone and side-chain ordering in proteins, we have performed multicanonical simulations of deka-peptide chains with various side groups. Glu(10), Gln(10), Asp(10), Asn(10), and Lys(10) were selected to cover a wide variety of possible interactions between the side chains of the monomers. All homopolymers undergo helix-coil transitions. We found that peptides with long side chains that are capable of hydrogen bonding, i.e., Glu(10), and Gln(10), exhibit a second transition at lower temperatures connected with side-chain ordering. This occurs in the gas phase as well as in solvent, although the character of the side-chain structure is different in each case. However, in polymers with short side chains capable of hydrogen bonding, i.e., Asp(10) and Asn(10), side-chain ordering takes place over a wide temperature range and exhibits no phase transition-like character. Moreover, non-backbone hydrogen bonds show enhanced formation and fluctuations already at the helix-coil transition temperature, indicating competition between side-chain and backbone hydrogen bond formation. Again, these results are qualitatively independent of the environment. Side-chain ordering in Lys(10), whose side groups are long and polar, also takes place over a wide temperature range and exhibits no phase transition-like character in both environments. Reasons for the observed chain length threshold and consequences from these results for protein folding are discussed. PMID- 17407348 TI - Improved non-chromatographic purification of a recombinant protein by cationic elastin-like polypeptides. AB - This paper reports an improvement in the purification of thioredoxin (Trx) expressed from E. coli by inverse transition cycling (ITC) using cationic elastin like polypeptides (ELPs). Two ELP libraries having 2% and 5% lysine residues and molecular weights ranging from 4 to 61.1 kDa showed greater salt sensitivity in their inverse transition behavior than purely aliphatic ELPs. Expression yield of Trx-ELP fusions was an unpredictable function of guest residue composition, but reducing the molecular weight of the ELP tag generally increased Trx yield. A cationic 4.3 kDa ELP is the shortest ELP used to purify any protein by ITC to date. A 15.9 kDa ELP with a guest residue composition of K:V:F of 1:7:1 was found to be the optimal cationic tag to purify Trx, as it provided 50% greater Trx yield and only required one-fifth the added NaCl for purification of Trx as compared to previously used aliphatic ELP tags. PMID- 17407349 TI - Binding of the environmental pollutant naphthol to bovine serum albumin. AB - The interactions between naphthol and bovine serum albumin (BSA) were investigated by spectroscopy. Our results prove the formation of complex between naphthol and BSA. Hydrophobic interaction dominates in the association reaction. The isomers stack with the aromatic residues in their binding sites with different geometries. Effects of BSA on the excited-state proton transfer and fluorescence spectra of the isomers indicate the different characters of their binding sites. 1-Naphthol inserts deeply into a hydrophobic cavity whereas 2 naphthol is in a basic environment on the surface of BSA. Naphthol statically quenches the fluorescence of BSA in a concentration-dependent manner positively deviating from the linear Stern-Volmer equation. Naphthol binds near Trp-134 in the subdomain IA of the native BSA and is accessible to Trp-212 when BSA is unfolded by naphthol. The folding pattern of the main chain is altered at high naphthol concentration as revealed by the change in the secondary structure. The binding of 1-naphthol is more cooperative than that of 2-naphthol. The extent of cooperativity was estimated by the Hill equation. PMID- 17407350 TI - Hydration of gluten: a dielectric, calorimetric, and fourier transform infrared study. AB - The nature of the hydration of proteins and the subsequent implications for functionality is a matter of importance in both pharmaceutical and food applications. Most published studies rely on the use of one technique and attempt to characterize the system. Few studies have used combinations of techniques. In this paper we report on the use of infrared, dielectric, and calorimetric methods to examine the hydration process of wheat gluten. This has been the subject of considerable study by other techniques and has been well characterized by our group. Results show that in both the infrared and dielectric measurements there is a change in behavior at about 35% water content. This is also the water content below which lowering the temperature of the sample does not result in ice formation. We suggest that at this water content the protein amide groups are fully hydrated, and beyond this point addition of water results in protein dilution rather than further hydration. PMID- 17407351 TI - Strongylophorine-26, an inhibitor of cancer cell Invasion: SAR revealed by synthesis of analogues. AB - The absolute configuration of strongylophorine-26 (1) was determined to be 4S, 5R, 8R, 9S, 10S, 13S, 14S by single-crystal X-ray diffraction analysis of the derivative 7 prepared from the co-occurring metabolite strongylophorine-8 (4) and chemical interconversion to the bislactone 8. Synthetic analogues (+)- and (-)-3 have been prepared in order to explore the structure-activity relationship for the anti-invasion pharmacophore of stronglylophorine-26. These studies revealed the unanticipated importance of the A ring lactone moiety for the anti-invasion activity of 1. PMID- 17407352 TI - Oligomeric benzylsulfonium salts: facile benzylation via high-load ROMP reagents. AB - The development of high-load, oligomeric benzylsulfonium salts, generated via ring-opening metathesis polymerization, and their utility in facile benzylations of various nucleophiles is reported. These oligomeric sulfonium salts exist as free-flowing powders and are stable at room temperature. After the benzylation event, purification is attained via simple dry load/filtration, followed by solvent removal to deliver products in excellent yield and purity. PMID- 17407353 TI - Asymmetric thio-Michael/nucleophilic addition domino reaction with chiral N sulfinimines. AB - Optically active N-sulfinimines underwent stereoselective Michael/nucleophilic addition domino reaction triggered by magnesium thiolate to give alpha phenylthiomethyl-beta-(N-sulfinylamino) esters in high diastereomeric excess. The adducts were readily converted into optically active alpha-methylene-beta-(N sulfinylamino)esters so that this reaction provides a useful asymmetric aza Baylis-Hillman-equivalent method. PMID- 17407354 TI - An improved understanding of the reaction of bis(bromomethyl)quinoxaline 1-N oxides with amines using substituent effects. AB - The reaction of bis(bromomethyl)quinoxaline N-oxides with amines is interesting from a reaction mechanism perspective and due to the reported biological activity of compounds in this general class. The complex mechanism of this reaction (particularly in the case of primary amines) is complicated further when C6 or C7 substituted mono-N-oxides are considered. In this study, the synthesis and subsequent characterization of a series of 2,3-bis(bromomethyl)quinoxaline 1-N oxides is reported. Experimental and computational evidence is used to show that the observed product ratios from the reaction with diethylamine reflect the influence of both the C6/C7 substituent and the N-oxide functional group on the initial nucleophilic substitution reaction. PMID- 17407355 TI - Spectroscopic and kinetic evidence for an accumulating intermediate in an SNV reaction with amine nucleophiles. Reaction of methyl beta-methylthio-alpha nitrocinnamate with piperidine and morpholine. AB - A spectroscopic and kinetic study of the reaction of methyl beta-methylthio-alpha nitrocinnamate (4-SMe) with morpholine, piperidine, and hydroxide ion in 50% DMSO/50% water (v/v) at 20 degrees C is reported. The reactions of 4-SMe with piperidine in a pH range from 10.12 to 11.66 and those with morpholine at pH 12.0 are characterized by two kinetic processes when monitored at lambdamax (364 nm) of the substrate, but by only one process when monitored at lambdamax (388) nm of the product. The rate constants obtained at 388 nm were the same as those determined for the slower of the two processes at 364 nm. These rate constants refer to product formation, whereas the faster process observed at 364 nm is associated with the loss of reactant to form an intermediate. In contrast, for the reaction of 4-SMe with morpholine at pH 8.62 the rates of product formation and disappearance of the substrate were the same, i.e., there is no accumulation of an intermediate. Likewise, the reaction of 4-SMe with OH- did not yield a detectable intermediate. The factors that allow the accumulation of intermediates in certain SNV reactions but not in others are discussed in detail, and structure reactivity comparisons are made with reactions of piperidine and morpholine with other highly activated vinylic substrates. PMID- 17407356 TI - Indium-mediated synthesis of heterobiaryls. AB - The palladium-mediated coupling reaction between triorganoindium reagents and organic electrophiles is extended to the synthesis of heteroaromatic compounds. Both electron-rich and electron-poor heterocycles can act as the organic electrophile or as the organoindium derivative. PMID- 17407357 TI - Homoallyl-cyclopropylcarbinyl cation manifold. Trimethylsilyl versus aryl stabilization. AB - A series of E- and Z-1-aryl-5-trimethylsilyl-3-buten-1-yl trifluoroacetates were solvolyzed in CD3CO2D, and rates of reaction as well as products derived from these reactions were determined. Hammett plots showed a break, which was indicative of a mechanistic change from a kC process when the most electron donating substituents were attached to the aryl group to a kDelta process involving formation of cyclized beta-silyl carbocation intermediates for electron withdrawing groups. In the case of p-CH3O substitution (a kC extreme), the cationic intermediate captures solvent (95%) or loses a proton (5%). In the case of m-CF3 substitution (a kDelta extreme), the beta-silyl cation intermediate desilylates to give vinylcyclopropane products. Substituents with intermediate electronic properties give more complex product mixtures. Solvolysis of pure Z trifluoroacetate (p-CH3) gives small amounts of E-trifluoroacetate (p-CH3) along with the E-substitution product. This isomerization suggests that the cyclized beta-silyl cation can isomerize and then reopen to a classical aryl-stabilized cation. By way of contrast, B3LYP/6-31G* computational studies show only cyclized beta-silyl cations as energy minima. Open kC cations are higher-energy nonminimum energy structures. PMID- 17407358 TI - Synthesis of liquid crystalline 4H-benzo[1,2,4]thiadiazines and generation of persistent radicals. AB - Four substituted 4H-benzo[1,2,4]thiadiazines 2 were prepared by condensation of the appropriate anilines and benzonitriles followed by oxidative cyclization. The preparation of three fluorinated derivatives 2b-2d proceeded smoothly, while the synthesis of 2a was problematic, presumably due to the relatively high electron density of the benzene ring. The four-ring derivatives 2c and 2d exhibited liquid crystalline properties (2c: Cr 95 SmA 158 I and 2d: Cr 142 SmA 212 I). 4H Benzo[1,2,4]thiadiazines 2 were oxidized with AgO to generate the corresponding persistent radicals 1 (g=2.0057). The stability of the radicals followed the order 1b approximately 1d>1c>1a, and the two fluorinated radicals 1b and 1d were isolated as crude solids. The lower stability of 1c is presumably due to the presence of the reactive benzylic CH position, and 1a lacks the stabilizing effect of the three fluorine atoms. ESR spectra for 1 were simulated using DFT derived hfcc as the starting point. PMID- 17407359 TI - Green and efficient synthesis of bidentate Schiff base Ru catalysts for olefin metathesis. AB - A simple, green synthetic protocol to bidentate Schiff base substituted ruthenium carbene complexes is described. These complexes are potent catalysts for olefin metathesis, but the original synthetic route employs the Schiff base ligands in the form of highly toxic thallium(I) salts. The new protocol involves a two-step, quasi-one-pot approach and silver(I) carbonate as base. The target ruthenium carbene complexes are obtained in yields that are similar to, or better than, those of the original protocol. PMID- 17407360 TI - Regioselective alkenylation of aromatic ketones with alkenylboronates using a RuH2(CO)(PPh3)3 catalyst via carbon-hydrogen bond cleavage. AB - The ruthenium-catalyzed alkenylation of C-H bonds with alkenylboronates has been explored for a series of aromatic ketones. The coupling reaction of pivalophenone (1) with 2-isopropenyl-5,5-dimethyl[1,3,2]dioxaborinane (2) gave the corresponding isopropenylation product in 73% yield. In the case of the reaction of a sterically congested alkenylboronate, such as 2-methylpropenylboronate (8), the yield was decreased slightly. When beta-styrylboronates were used, the corresponding coupling products were obtained in good yields. The reaction of acetophenone with alpha-styrylboronate afforded the corresponding 1:1 coupling product, exclusively. PMID- 17407361 TI - Combinatorial discovery of reusable noncovalent supports for enzyme immobilization and nonaqueous catalysis. AB - A simple and effective method is described for the preparation of enzyme containing materials that possess excellent catalytic activity, mechanical strength, and reusability. Uniform spherical beads were produced via the colyophilization of alpha-chymotrypsin with the support materials, leaving the active enzyme entrapped within the porous "ice-templated" support matrix. The composites were assayed for catalytic activity by monitoring a nonaqueous transesterification reaction. The mechanical strength for each composite was measured using a compression assay. Initial screens identified a set of six support materials that contributed favorably to either the enzyme activity or to the mechanical strength of the composite. A design of experiments (DoE) methodology was employed to screen 80 combinations of these six "base" materials. A model representing this formulation space was constructed which could be used to predict both the catalytic activity and mechanical strength with reasonable accuracy for any combination of the six base component materials. This model was used to predict optimized materials with an enzyme activity that was 50 times greater than that of the free enzyme. The model was also used to set a minimum acceptable mechanical stability for these composites, and the resulting materials were shown to be reusable for at least ten reaction cycles. PMID- 17407362 TI - Toward self-assembled ferroelectric random access memories: hard-wired switching capacitor arrays with almost Tb/in.(2) densities. AB - We report on the successful fabrication of arrays of switchable nanocapacitors made by harnessing the self-assembly of materials. The structures are composed of arrays of 20-40 nm diameter Pt nanowires, spaced 50-100 nm apart, electrodeposited through nanoporous alumina onto a thin film lower electrode on a silicon wafer. A thin film ferroelectric (both barium titanate (BTO) and lead zirconium titanate (PZT)) has been deposited on top of the nanowire array, followed by the deposition of thin film upper electrodes. The PZT nanocapacitors exhibit hysteresis loops with substantial remnant polarizations, while although the switching performance was inferior, the low-field characteristics of the BTO nanocapacitors show dielectric behavior comparable to conventional thin film heterostructures. While registration is not sufficient for commercial RAM production, this is nevertheless an embryonic form of the highest density hard wired FRAM capacitor array reported to date and compares favorably with atomic force microscopy read-write densities. PMID- 17407363 TI - Pharmacologic management of glaucoma in childhood. AB - An evidence-based review of the drugs available for the medical management of childhood glaucoma is presented; almost all of the drugs are not licensed for use in children. Despite this, most topical drugs are safe; however, there are some significant exceptions, such as brimonidine, which may cause apnea, among other life-threatening adverse events, in infants. Broad families of drugs are available including topical adrenoceptor antagonists, topical and systemic carbonic anhydrase inhibitors, prostaglandin analogs, adrenoceptor agonists, parasympathomimetics, and combination preparations. These drugs help to reduce intraocular pressure by reducing aqueous production or increasing the outflow facility. The variety of anti-ocular hypertensive medications for childhood glaucoma has increased in recent years. The vast majority of data on these medications are from adult studies but each year more experience of their use in pediatric glaucoma is gained. In general, topical treatment is well tolerated; however, the prescribing clinician and carers should be aware of potential adverse effects and how they may present. PMID- 17407364 TI - Emerging treatments and gene expression profiling in high-risk medulloblastoma. AB - The past decades have seen an increase in the survival rates of patients with standard-risk medulloblastoma. Efforts have, therefore, been focused on obtaining better results in the treatment of patients with high-risk tumors. In addition to consolidated therapies, novel approaches such as small molecules, monoclonal antibodies, and antiangiogenic therapies that aim to improve outcomes and quality of life are now available through new breakthroughs in the molecular biology of medulloblastoma. The advent of innovative anticancer drugs tested in brain tumors has important consequences for personalized therapy. Gene expression profiling of medulloblastoma can be used to identify the genes and signaling transduction pathways that are crucial for the tumorigenesis process, thereby revealing both new targets for therapy and sensitive/resistance phenotypes. The interpretation of microarray data for new treatments of patients with high-risk medulloblastoma, as well as other poor prognosis tumors, should be developed through a consensus multidisciplinary approach involving oncologists, neurosurgeons, radiotherapists, biotechnologists, bioinformaticists, and other professionals. PMID- 17407365 TI - Serotonin reuptake inhibitor-induced perinatal complications. AB - There are a growing number of concerns about the utilization of serotonin reuptake inhibitors (SRIs) in late pregnancy and the onset of perinatal complications. This review aimed to analyze and summarize the studies evaluating the risk of perinatal complications (such as low birth weight, preterm delivery, withdrawal or toxic phenomena, and other detrimental events/poor neonatal outcomes) related to maternal SRI use in late pregnancy. A computerized search of MEDLINE (1966-January 2007) and PsycINFO (1974-January 2007) databases was performed. Articles describing perinatal complications after late in utero exposure to SRIs were selected and also reviewed for additional references. Fifty studies met the inclusion criteria. Exposure to SRIs late in pregnancy is clearly associated with an increased risk of infants developing a constellation of symptoms, including CNS and respiratory effects, often requiring close infant observation and supportive or specific treatment in intensive care units. Such symptoms are not always due to toxic or withdrawal reactions. Indeed, some evidence suggests that SRIs may interfere with the physiology of the respiratory system and parasympathetic activity in neonates. Of the most methodologically relevant studies reviewed, 50% have been published in the last 3 years. Hence, it is possible that further concerning data will become available in the future. For these reasons, the opportunity of tapering and discontinuing SRIs in late pregnancy should be taken into consideration, although to date the evidence to support such a clinical decision is preliminary. PMID- 17407366 TI - Immunomodulatory effects of macrolide antibiotics in respiratory disease: therapeutic implications for asthma and cystic fibrosis. AB - The macrolide antibiotics are a family of related 14- or 15-membered lactone ring antibiotics. There has been recent interest in the beneficial effects of these drugs as immune modulators in respiratory conditions in children. Cystic fibrosis (CF) and asthma, both of which occur in childhood, have an underlying inflammatory component and are associated with significant morbidity. The pathogenesis of both conditions is poorly understood but several molecular mechanisms have been suggested. In CF, these mechanisms broadly involve altered chloride transport and alteration of the airway surface liquid with disordered neutrophilic inflammation. There is much evidence for a proinflammatory propensity in CF immune effector and epithelial cells and many studies indicate that macrolides modulate these inflammatory processes. Recent studies have confirmed a clinical improvement in CF following treatment with macrolides, but the exact mechanisms by which they work are unknown. Asthma is likely to represent several different phenotypes but in all of these, airway obstruction, bronchial hyperresponsiveness, and inflammation are central processes. Results from trials using macrolides have suggested an improvement in clinical outcome. The putative mechanisms of macrolide immunomodulatory action include improvement of the primary defense mechanisms, inhibition of the bacteria-epithelial cell interaction, modulation of the signaling pathway and chemokine release, and direct neutrophil effects. Putative mechanisms of phenotypic modulation have also been proposed involving interactions with nitric oxide, endothelin-1, and bronchoconstriction, endothelial growth factors and airway remodeling, and bioactive phospholipids in both CF and asthma. Further characterization of these effects and development of targeted designer drugs will further expand our therapeutic repertoire and lead to improved quality and quantity of life for patients with CF and asthma. PMID- 17407367 TI - Pharmacokinetics of cefprozil in plasma and middle ear fluid: in children undergoing treatment for acute otitis media. AB - BACKGROUND: Despite the wide-scale use of cefprozil for acute otitis media (AOM), there are only limited data available regarding the pharmacokinetic profile of this agent in the pediatric population. OBJECTIVE: To characterize the plasma and middle ear fluid (MEF) pharmacokinetic profile of cefprozil in pediatric patients with AOM. METHODS: Pharmacokinetic sampling was obtained as part of a phase IV, multicenter, open-label study of children with AOM receiving cefprozil suspension 15 mg/kg twice daily. A single blood sample was obtained 4-6 days after the initiation of cefprozil therapy and a simultaneous MEF sample was obtained by tympanocentesis when clinically indicated. Cefprozil concentrations in both matrices were determined using a validated high-performance liquid chromatography methodology. A composite profile of cefprozil concentration data in each matrix was constructed and values for the pharmacokinetic parameters were obtained using conventional modeling techniques. RESULTS: Plasma concentrations were obtained in 53 children aged 6-48 months. In this population the maximum concentration (C(max)) in plasma was 9.18 microg/mL, the time to C(max) (t(max)) was 1.5 hours, and the terminal elimination half-life (t((1/2))(beta)) was 0.98 hours. Simultaneous plasma and MEF concentration data were available in 22 children. In this subset the C(max) in plasma was 8.2 microg/mL, the t(max) was 1.9 hours, and the t((1/2))(beta) was 1.02 hours; the corresponding MEF C(max) was 2.4 microg/mL, the t(max) was 3.5 hours, and the t((1/2))(beta) was 1.23 hours. Cefprozil MEF penetration as assessed using the ratio of the area under the concentration-time curves from the two matrices was 28%. Moreover, concentrations in MEF approximated 1 microg/mL 6 hours' post-dose. CONCLUSIONS: The plasma profile of cefprozil in the current analysis is consistent with previously reported values in children receiving the 15 mg/kg twice daily dose. MEF penetration and the duration of drug exposure at the site of infection support the clinical utility of this agent for organisms with minimum inhibitory concentrations (MIC) of < or =1 microg/mL. However, these results also predict higher clinical failure when using this dose of cefprozil against penicillin-non susceptible Streptococcus pneumoniae or Haemophilus influenzae because of typically higher MIC values for these organisms. PMID- 17407368 TI - Generalized seizures following topical lidocaine administration during circumcision: establishing causation. AB - We report a case of neonatal seizures after lidocaine administration for circumcision. A 3-month-old male infant received an overdose as evidenced by toxic lidocaine levels and developed generalized seizures shortly after. Back extrapolation of the serum lidocaine concentration to time zero was used to determine the administered dose. The Naranjo scale was used to determine causation; probable causation was defined. Particular care must be taken to administer an appropriate dose of local anesthetics in infants to avoid life threatening seizures. PMID- 17407369 TI - Lisdexamfetamine. AB - Lisdexamfetamine is an amphetamine prodrug, comprising an l-lysine amino acid covalently bonded to dextroamphetamine (d-amphetamine). Lisdexamfetamine is approved in the US for the treatment of attention-deficit hyperactivity disorder in children aged 6-12 years. Lisdexamfetamine is a therapeutically inactive molecule. After oral ingestion, lisdexamfetamine is hydrolyzed to l-lysine, a naturally occurring essential amino acid, and active d-amphetamine, which is responsible for the activity of the drug. In a well designed pharmacodynamic study in adult stimulant abusers, 50 or 100 mg doses of oral lisdexamfetamine had less likability than d-amphetamine 40 mg, suggesting a reduced abuse potential. Through rate-limited hydrolysis in the body, l-lysine is cleaved, gradually releasing pharmacologically active d-amphetamine. The pharmacokinetics of lisdexamfetamine suggest a reduced potential for abuse. In two well designed trials in children aged 6-12 years with attention-deficit hyperactivity disorder (ADHD), the efficacy of lisdexamfetamine was superior to that of placebo in improving symptoms associated with ADHD. Adverse events with lisdexamfetamine were, in general, mild to moderate in severity and consistent with those commonly reported with amphetamine. PMID- 17407380 TI - Understanding the evolutionary fate of finite populations: the dynamics of mutational effects. AB - The most consistent result in more than two decades of experimental evolution is that the fitness of populations adapting to a constant environment does not increase indefinitely, but reaches a plateau. Using experimental evolution with bacteriophage, we show here that the converse is also true. In populations small enough such that drift overwhelms selection and causes fitness to decrease, fitness declines down to a plateau. We demonstrate theoretically that both of these phenomena must be due either to changes in the ratio of beneficial to deleterious mutations, the size of mutational effects, or both. We use mutation accumulation experiments and molecular data from experimental evolution to show that the most significant change in mutational effects is a drastic increase in the rate of beneficial mutation as fitness decreases. In contrast, the size of mutational effects changes little even as organismal fitness changes over several orders of magnitude. These findings have significant implications for the dynamics of adaptation. PMID- 17407381 TI - Intramolecular and intermolecular interactions of protein kinase B define its activation in vivo. AB - Protein kinase B (PKB/Akt) is a pivotal regulator of diverse metabolic, phenotypic, and antiapoptotic cellular controls and has been shown to be a key player in cancer progression. Here, using fluorescent reporters, we shown in cells that, contrary to in vitro analyses, 3-phosphoinositide-dependent protein kinase 1 (PDK1) is complexed to its substrate, PKB. The use of Forster resonance energy transfer detected by both frequency domain and two-photon time domain fluorescence lifetime imaging microscopy has lead to novel in vivo findings. The preactivation complex of PKB and PDK1 is maintained in an inactive state through a PKB intramolecular interaction between its pleckstrin homology (PH) and kinase domains, in a "PH-in" conformer. This domain-domain interaction prevents the PKB activation loop from being phosphorylated by PDK1. The interactive regions for this intramolecular PKB interaction were predicted through molecular modeling and tested through mutagenesis, supporting the derived model. Physiologically, agonist-induced phosphorylation of PKB by PDK1 occurs coincident to plasma membrane recruitment, and we further shown here that this process is associated with a conformational change in PKB at the membrane, producing a "PH-out" conformer and enabling PDK1 access the activation loop. The active, phosphorylated, "PH-out" conformer can dissociate from the membrane and retain this conformation to phosphorylate substrates distal to the membrane. These in vivo studies provide a new model for the mechanism of activation of PKB. This study takes a crucial widely studied regulator (physiology and pathology) and addresses the fundamental question of the dynamic in vivo behaviour of PKB with a detailed molecular mechanism. This has important implications not only in extending our understanding of this oncogenic protein kinase but also in opening up distinct opportunities for therapeutic intervention. PMID- 17407382 TI - Survey sequencing and comparative analysis of the elephant shark (Callorhinchus milii) genome. AB - Owing to their phylogenetic position, cartilaginous fishes (sharks, rays, skates, and chimaeras) provide a critical reference for our understanding of vertebrate genome evolution. The relatively small genome of the elephant shark, Callorhinchus milii, a chimaera, makes it an attractive model cartilaginous fish genome for whole-genome sequencing and comparative analysis. Here, the authors describe survey sequencing (1.4x coverage) and comparative analysis of the elephant shark genome, one of the first cartilaginous fish genomes to be sequenced to this depth. Repetitive sequences, represented mainly by a novel family of short interspersed element-like and long interspersed element-like sequences, account for about 28% of the elephant shark genome. Fragments of approximately 15,000 elephant shark genes reveal specific examples of genes that have been lost differentially during the evolution of tetrapod and teleost fish lineages. Interestingly, the degree of conserved synteny and conserved sequences between the human and elephant shark genomes are higher than that between human and teleost fish genomes. Elephant shark contains putative four Hox clusters indicating that, unlike teleost fish genomes, the elephant shark genome has not experienced an additional whole-genome duplication. These findings underscore the importance of the elephant shark as a critical reference vertebrate genome for comparative analysis of the human and other vertebrate genomes. This study also demonstrates that a survey-sequencing approach can be applied productively for comparative analysis of distantly related vertebrate genomes. PMID- 17407383 TI - Forest elephant crisis in the Congo Basin. AB - Debate over repealing the ivory trade ban dominates conferences of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). Resolving this controversy requires accurate estimates of elephant population trends and rates of illegal killing. Most African savannah elephant populations are well known; however, the status of forest elephants, perhaps a distinct species, in the vast Congo Basin is unclear. We assessed population status and incidence of poaching from line-transect and reconnaissance surveys conducted on foot in sites throughout the Congo Basin. Results indicate that the abundance and range of forest elephants are threatened from poaching that is most intense close to roads. The probability of elephant presence increased with distance to roads, whereas that of human signs declined. At all distances from roads, the probability of elephant occurrence was always higher inside, compared to outside, protected areas, whereas that of humans was always lower. Inside protected areas, forest elephant density was correlated with the size of remote forest core, but not with size of protected area. Forest elephants must be prioritised in elephant management planning at the continental scale. PMID- 17407384 TI - Addressing the global neglect of childhood hearing impairment in developing countries. PMID- 17407385 TI - Interrupted time-series analysis of regulations to reduce paracetamol (acetaminophen) poisoning. AB - BACKGROUND: Paracetamol (acetaminophen) poisoning is the leading cause of acute liver failure in Great Britain and the United States. Successful interventions to reduced harm from paracetamol poisoning are needed. To achieve this, the government of the United Kingdom introduced legislation in 1998 limiting the pack size of paracetamol sold in shops. Several studies have reported recent decreases in fatal poisonings involving paracetamol. We use interrupted time-series analysis to evaluate whether the recent fall in the number of paracetamol deaths is different to trends in fatal poisoning involving aspirin, paracetamol compounds, antidepressants, or nondrug poisoning suicide. METHODS AND FINDINGS: We calculated directly age-standardised mortality rates for paracetamol poisoning in England and Wales from 1993 to 2004. We used an ordinary least-squares regression model divided into pre- and postintervention segments at 1999. The model included a term for autocorrelation within the time series. We tested for changes in the level and slope between the pre- and postintervention segments. To assess whether observed changes in the time series were unique to paracetamol, we compared against poisoning deaths involving compound paracetamol (not covered by the regulations), aspirin, antidepressants, and nonpoisoning suicide deaths. We did this comparison by calculating a ratio of each comparison series with paracetamol and applying a segmented regression model to the ratios. No change in the ratio level or slope indicated no difference compared to the control series. There were about 2,200 deaths involving paracetamol. The age-standardised mortality rate rose from 8.1 per million in 1993 to 8.8 per million in 1997, subsequently falling to about 5.3 per million in 2004. After the regulations were introduced, deaths dropped by 2.69 per million (p = 0.003). Trends in the age standardised mortality rate for paracetamol compounds, aspirin, and antidepressants were broadly similar to paracetamol, increasing until 1997 and then declining. Nondrug poisoning suicide also declined during the study period, but was highest in 1993. The segmented regression models showed that the age standardised mortality rate for compound paracetamol dropped less after the regulations (p = 0.012) but declined more rapidly afterward (p = 0.031). However, age-standardised rates for aspirin and antidepressants fell in a similar way to paracetamol after the regulations. Nondrug poisoning suicide declined at a similar rate to paracetamol after the regulations were introduced. CONCLUSIONS: Introduction of regulations to limit availability of paracetamol coincided with a decrease in paracetamol-poisoning mortality. However, fatal poisoning involving aspirin, antidepressants, and to a lesser degree, paracetamol compounds, also showed similar trends. This raises the question whether the decline in paracetamol deaths was due to the regulations or was part of a wider trend in decreasing drug-poisoning mortality. We found little evidence to support the hypothesis that the 1998 regulations limiting pack size resulted in a greater reduction in poisoning deaths involving paracetamol than occurred for other drugs or nondrug poisoning suicide. PMID- 17407386 TI - Gene expression signatures that predict radiation exposure in mice and humans. AB - BACKGROUND: The capacity to assess environmental inputs to biological phenotypes is limited by methods that can accurately and quantitatively measure these contributions. One such example can be seen in the context of exposure to ionizing radiation. METHODS AND FINDINGS: We have made use of gene expression analysis of peripheral blood (PB) mononuclear cells to develop expression profiles that accurately reflect prior radiation exposure. We demonstrate that expression profiles can be developed that not only predict radiation exposure in mice but also distinguish the level of radiation exposure, ranging from 50 cGy to 1,000 cGy. Likewise, a molecular signature of radiation response developed solely from irradiated human patient samples can predict and distinguish irradiated human PB samples from nonirradiated samples with an accuracy of 90%, sensitivity of 85%, and specificity of 94%. We further demonstrate that a radiation profile developed in the mouse can correctly distinguish PB samples from irradiated and nonirradiated human patients with an accuracy of 77%, sensitivity of 82%, and specificity of 75%. Taken together, these data demonstrate that molecular profiles can be generated that are highly predictive of different levels of radiation exposure in mice and humans. CONCLUSIONS: We suggest that this approach, with additional refinement, could provide a method to assess the effects of various environmental inputs into biological phenotypes as well as providing a more practical application of a rapid molecular screening test for the diagnosis of radiation exposure. PMID- 17407387 TI - Macrosomia and hyperinsulinaemic hypoglycaemia in patients with heterozygous mutations in the HNF4A gene. AB - BACKGROUND: Macrosomia is associated with considerable neonatal and maternal morbidity. Factors that predict macrosomia are poorly understood. The increased rate of macrosomia in the offspring of pregnant women with diabetes and in congenital hyperinsulinaemia is mediated by increased foetal insulin secretion. We assessed the in utero and neonatal role of two key regulators of pancreatic insulin secretion by studying birthweight and the incidence of neonatal hypoglycaemia in patients with heterozygous mutations in the maturity-onset diabetes of the young (MODY) genes HNF4A (encoding HNF-4alpha) and HNF1A/TCF1 (encoding HNF-1alpha), and the effect of pancreatic deletion of Hnf4a on foetal and neonatal insulin secretion in mice. METHODS AND FINDINGS: We examined birthweight and hypoglycaemia in 108 patients from families with diabetes due to HNF4A mutations, and 134 patients from families with HNF1A mutations. Birthweight was increased by a median of 790 g in HNF4A-mutation carriers compared to non mutation family members (p < 0.001); 56% (30/54) of HNF4A-mutation carriers were macrosomic compared with 13% (7/54) of non-mutation family members (p < 0.001). Transient hypoglycaemia was reported in 8/54 infants with heterozygous HNF4A mutations, but was reported in none of 54 non-mutation carriers (p = 0.003). There was documented hyperinsulinaemia in three cases. Birthweight and prevalence of neonatal hypoglycaemia were not increased in HNF1A-mutation carriers. Mice with pancreatic beta-cell deletion of Hnf4a had hyperinsulinaemia in utero and hyperinsulinaemic hypoglycaemia at birth. CONCLUSIONS: HNF4A mutations are associated with a considerable increase in birthweight and macrosomia, and are a novel cause of neonatal hypoglycaemia. This study establishes a key role for HNF4A in determining foetal birthweight, and uncovers an unanticipated feature of the natural history of HNF4A-deficient diabetes, with hyperinsulinaemia at birth evolving to decreased insulin secretion and diabetes later in life. PMID- 17407388 TI - Type 2 diabetes: hypoinsulinism, hyperinsulinism, or both? PMID- 17407389 TI - Does restricting pack size of paracetamol (acetaminophen) reduce suicides? PMID- 17407390 TI - Effects of a step-therapy program for angiotensin receptor blockers on antihypertensive medication utilization patterns and cost of drug therapy. AB - BACKGROUND: Step therapy for angiotensin receptor blockers (ARBs) requiring prior use of angiotensin-converting enzyme inhibitors (ACEIs) is a common cost containment intervention in managed care. OBJECTIVE: This study was designed to assess the effectiveness of the step-therapy intervention for ARBs, including ARB/hydrochlorthiazide (HCTZ) combinations, as measured by prescription use patterns and antihypertensive drug ingredient costs. METHODS: Rejected and paid pharmacy claims data were evaluated from 3 health plans with a total membership of approximately 1 million. These plans had implemented a step-therapy intervention for ARBs from May 1, 2001, through February 28, 2003. Patients in the intervention group who had experienced a claim rejection for an ARB within the first 6 months of program implementation (i.e., had had no ACEI [or ACEI/HCTZ combination] or ARB [or ARB/HCTZ] claim in the preceding 3 months) were followed for 1 year after the ARB claim rejection. The rate of initiation of ARB versus ACEI and other outcomes was compared with similar data from a health plan with approximately 2 million members that did not have a step-therapy intervention for ARBs (comparison group). Mean and median total antihypertensive drug ingredient costs per patient and per day of therapy over 12 months were analyzed for the intervention and comparison groups. One pharmacy benefit manager administered the pharmacy benefits for the intervention and comparison health plans during the entire study period from May 1, 2001, through February 28, 2004, and the drug formulary was similar for all health plans. RESULTS: In the step-therapy health plans, before the criterion for 15 months of continuous eligibility was applied, there were 8,904 patients (approximately 0.9% of health plan members) who either attempted and were rejected for an ARB or who newly started ACEI therapy, compared with 44,788 patients (approximately 2.2% of members in the comparison health plan) who newly started ARB or ACEI therapy without the step-therapy intervention. After the eligibility criterion was applied, there were 6,758 intervention health plan members (0.7% of members) and 33,709 comparison health plan members (1.7% of members) in the 2 study groups. In addition to the smaller proportion of total members affected by the intervention in the ARB step-therapy health plans, a smaller proportion of ARB/ACEI patients attempted to obtain an ARB (1,296/6,758 or 19.2%) compared with the health plan without step therapy (8,697/33,709 or 25.8%, P <0.001). Of the 1,296 patients who attempted to obtain an ARB and were rejected in the step-therapy group, 578 patients (44.6%) went through the prior-authorization process and received an ARB as initial therapy, 632 patients (48.8%) received other antihypertensive therapy, and 86 patients (6.6%) did not receive any antihypertensive therapy within the 12-month follow-up period. In the 12 months of follow-up, 51.1% (323/632) of patients in the intervention group who received other antihypertensives as index therapy switched to or added an ARB, and 1,234 of total ACE/ARB patients (n = 6,758, 18.3%) received ARB therapy in the health plan with step therapy compared with 10,498 of 33,709 total ACEI/ARB patients (31.1%) who received ARB therapy in the health plan without step therapy. The mean antihypertensive drug cost per patient was lower in the intervention group ($370.00) than in the comparison group ($445.12; P <0.001), and the average cost per day of antihypertensive drug therapy was 12.8% lower in the step-therapy group ($0.82) than in the comparison group ($0.94). Unadjusted annual cost savings were $75.12 per patient, and ordinary least squares regression analysis showed that the ARB step-therapy intervention was associated with $43.91 in antihypertensive drug cost savings per patient over 12 months. CONCLUSIONS: Within 12 months of follow-up, a step-therapy intervention for ARBs was associated with an 18% ratio of ARB users to total ACEI/ARB users compared with a 31% ratio in a comparison health plan without the ARB step-therapy intervention. Approximately 45% of patients who did not receive an ARB as a result of the step-therapy intervention had either switched to or added an ARB within 12 months of the intervention, and almost 7% of patients did not receive any antihypertensive therapy. Antihypertensive drug cost was about 13% lower for the ACEI/ARB patients in the intervention group, creating approximately $368,000 in savings in 1 year or $0.03 per member per month across the 1 million health plan members. PMID- 17407391 TI - Cost-effectiveness of four immunomodulatory therapies for relapsing-remitting multiple sclerosis: a Markov model based on long-term clinical data. AB - BACKGROUND: Before the introduction of the immunomodulatory therapies for multiple sclerosis (MS), treatment options for MS consisted of symptomatic management (physical therapy and pharmacological treatment for symptom management). Symptomatic management for MS has been supplemented in the past decade by 2 new classes of immunomodulatory therapies that have been approved as first-line treatments for relapsing-remitting multiple sclerosis (RRMS): subcutaneous glatiramer acetate (SC GA) and 3 beta-interferons: intramuscular interferon beta-1a (IM IFNbeta-1a), SC IFNbeta-1a, and SC IFNbeta-1b. OBJECTIVE: To estimate the economic outcomes of 5 treatment strategies: symptom management alone, symptom management combined with SC GA, IM IFNbeta1-a, SC IFNbeta1-a, or SC IFNbeta1-b in patients diagnosed with RRMS. METHODS: A literature-based Markov model was developed to assess the cost-effectiveness of 5 treatment strategies for managing a hypothetical cohort of patients diagnosed with RRMS in the United States--4 immunomodulatory drug therapies and symptom management alone. Health states were based on the Kurtzke Expanded Disability Status Scale (EDSS), a widely accepted scale for assessing RRMS (higher EDSS scores = increased disease severity). Baseline relapse and disease progression transition probabilities for symptom management were obtained from natural history studies. Treatment effects of the immunomodulatory therapies were estimated by applying a percentage reduction to the symptom management transition probabilities for relapse (27% reduction) and disease progression (30% reduction). Transition probabilities were subsequently adjusted to account for (1) the effects of neutralizing antibodies, specifically on relapse rates by assuming no additional therapy benefits after the second year of continuous therapy, and (2) treatment discontinuation. Therapy specific data were obtained from clinical trials and long-term follow-up observational studies. Transitions among health states occurred in 1-month cycles for the lifetime of a patient. Costs (2005 US$) and outcomes were discounted at 3% annually. RESULTS: The incremental cost per quality-adjusted life-year for the 4 immunomodulatory therapies is $258,465, $303,968, $416,301, and $310,691 for SC GA, IM IFNbeta-1a, SC IFNbeta-1a, and SC IFNbeta-1b, respectively, compared with symptom management alone. Sensitivity analyses demonstrated that results were sensitive to changes in utilities, disease progression rates, time horizon, and immunomodulatory therapy cost. CONCLUSIONS: The pharmacoeconomic model determined that SC GA was the best strategy of the 4 immunomodulatory therapies used to manage MS and resulted in better outcomes than symptom management alone. Sensitivity analyses indicated that the model was sensitive to changes in a number of key parameters, and thus changes in these key parameters would likely influence the estimated cost-effectiveness results. Head-to-head randomized clinical trials comparing the immunomodulatory therapies for the treatment of MS are necessary to validate the projections from the pharmacoeconomic analyses, particularly since the results available today from the clinical trials do not account adequately for treatment dropouts. PMID- 17407392 TI - Actuarial analysis of private payer administrative claims data for women with endometriosis. AB - BACKGROUND: Endometriosis is a painful, chronic disease affecting 5.5 million women and girls in the United States and Canada and millions more worldwide. The usual age range of women diagnosed with endometriosis is 20 to 45 years. Endometriosis has an estimated prevalence of 10% among women of reproductive age, although estimates of prevalence vary greatly. Endometriosis is the most common gynecological cause of chronic pelvic pain, but published information on its associated medical care costs is scarce. OBJECTIVE: The aim of this study was to determine (1) the prevalence of endometriosis in the United States, (2) the amount of health care services used by women coded with endometriosis in a commercial medical claims database during 1999 to 2003, and (3) the endometriosis related costs for 2003, the most recent data available at the time the study was performed. METHODS: This study was a retrospective review of administrative data for commercial payers, which included enrollment, eligibility, and claims payment data contained in the Medstat Marketscan database for approximately 4 million commercial insurance members. All claims and membership data were extracted for each woman aged 18 to 55 years who had at least 1 medical or hospital claim with a diagnosis code for endometriosis (International Classification of Diseases, Ninth Revision, Clinical Modification [ICD-9-CM] codes 617.00-617.99) for 1999 through 2003. Claims data from 1999 through 2003 were used to determine prevalence and health care resource utilization (i.e., annual admission rate, annual surgical rate, distribution of endometriosis-related surgeries, and prevalence of comorbid conditions). The cost analysis was based on claims from 2003 only. Cost was defined as the payer-allowed charge, which equals the net payer cost plus member cost share. RESULTS: The prevalence of women with medical claims (inpatient and/or outpatient) containing ICD-9-CM codes for endometriosis was 1.1% for the age band of 30 to 39 years and 0.7% over the entire age span of 18 to 55 years. The medical costs per patient per month (PPPM) for women with endometriosis were 63% greater ($706 PPPM) than those of the average woman per member per month ($433) in 2003; inpatient hospital costs accounted for 32% of total direct medical costs. Between 1999 and 2003, these women with endometriosis who were identified by either inpatient and/or outpatient claims had high rates of hospital admission (53% for any reason; 38% for an endometriosis-related reason) and a high annual surgical procedure rate (64%). Additionally, women with endometriosis frequently suffered from comorbid conditions, and these conditions were associated with greater PPPM costs of 15% to 50% for women with an endometriosis diagnosis code, depending on the condition. Interstitial cystitis was associated with 50% greater cost ($1,061 PPPM); depression, 41% ($997 PPPM); migraine, 40% ($988 PPPM); irritable bowel syndrome, 34% ($943 PPPM); chronic fatigue syndrome, 29% ($913 PPPM); abdominal pain, 20% ($846 PPPM); and infertility, 15% ($813 PPPM). CONCLUSIONS: Women with endometriosis have a high hospital admission rate and surgical procedure rate and a high incidence of comorbid conditions. Consequently, these women incur total medical costs that are, on average, 63% higher than medical costs for the average woman in a commercially insured group. PMID- 17407393 TI - Assessing step-therapy programs:a step in the right direction. PMID- 17407394 TI - Medication therapy management programs: when will the outcomes come out? PMID- 17407395 TI - Expert opinion regarding clinical and other outcome considerations in the formulary review of immune globulin. PMID- 17407396 TI - Outcomes of sword swallowing and pharmaceutical step-therapy interventions. PMID- 17407397 TI - Pharmacoeconomic modeling of drug therapies for multiple sclerosis--are we building houses on sand? PMID- 17407398 TI - Manufacturer response to AMCP Format dossier requests. PMID- 17407399 TI - Correction necessary regarding CMS guidance for coverage of new multiple-source brand drugs. PMID- 17407400 TI - Interplanetary transfer of photosynthesis: an experimental demonstration of a selective dispersal filter in planetary island biogeography. AB - We launched a cryptoendolithic habitat, made of a gneissic impactite inoculated with Chroococcidiopsis sp., into Earth orbit. After orbiting the Earth for 16 days, the rock entered the Earth's atmosphere and was recovered in Kazakhstan. The heat of entry ablated and heated the rock to a temperature well above the upper temperature limit for life to below the depth at which light levels are insufficient for photosynthetic organisms ( approximately 5 mm), thus killing all of its photosynthetic inhabitants. This experiment shows that atmospheric transit acts as a strong biogeographical dispersal filter to the interplanetary transfer of photosynthesis. Following atmospheric entry we found that a transparent, glassy fusion crust had formed on the outside of the rock. Re-inoculated Chroococcidiopsis grew preferentially under the fusion crust in the relatively unaltered gneiss beneath. Organisms under the fusion grew approximately twice as fast as the organisms on the control rock. Thus, the biologically destructive effects of atmospheric transit can generate entirely novel and improved endolithic habitats for organisms on the destination planetary body that survive the dispersal filter. The experiment advances our understanding of how island biogeography works on the interplanetary scale. PMID- 17407401 TI - On biogenicity criteria for endolithic microborings on early Earth and beyond. AB - Micron-sized cavities created by the actions of rock-etching microorganisms known as euendoliths are explored as a biosignature for life on early Earth and perhaps Mars. Rock-dwelling organisms can tolerate extreme environmental stresses and are excellent candidates for the colonization of early Earth and planetary surfaces. Here, we give a brief overview of the fossil record of euendoliths in both sedimentary and volcanic rocks. We then review the current understanding of the controls upon the distribution of euendolithic microborings and use these to propose three lines of approach for testing their biogenicity: first, a geological setting that demonstrates a syngenetic origin for the euendolithic microborings; second, microboring morphologies and distributions that are suggestive of biogenic behavior and distinct from ambient inclusion trails; and third, elemental and isotopic evidence suggestive of biological processing. We use these criteria and the fossil record of terrestrial euendoliths to outline potential environments and techniques to search for endolithic microborings on Mars. PMID- 17407403 TI - A reappraisal of the habitability of planets around M dwarf stars. AB - Stable, hydrogen-burning, M dwarf stars make up about 75% of all stars in the Galaxy. They are extremely long-lived, and because they are much smaller in mass than the Sun (between 0.5 and 0.08 M(Sun)), their temperature and stellar luminosity are low and peaked in the red. We have re-examined what is known at present about the potential for a terrestrial planet forming within, or migrating into, the classic liquid-surface-water habitable zone close to an M dwarf star. Observations of protoplanetary disks suggest that planet-building materials are common around M dwarfs, but N-body simulations differ in their estimations of the likelihood of potentially habitable, wet planets that reside within their habitable zones, which are only about one-fifth to 1/50th of the width of that for a G star. Particularly in light of the claimed detection of the planets with masses as small as 5.5 and 7.5 M(Earth) orbiting M stars, there seems no reason to exclude the possibility of terrestrial planets. Tidally locked synchronous rotation within the narrow habitable zone does not necessarily lead to atmospheric collapse, and active stellar flaring may not be as much of an evolutionarily disadvantageous factor as has previously been supposed. We conclude that M dwarf stars may indeed be viable hosts for planets on which the origin and evolution of life can occur. A number of planetary processes such as cessation of geothermal activity or thermal and nonthermal atmospheric loss processes may limit the duration of planetary habitability to periods far shorter than the extreme lifetime of the M dwarf star. Nevertheless, it makes sense to include M dwarf stars in programs that seek to find habitable worlds and evidence of life. This paper presents the summary conclusions of an interdisciplinary workshop (http://mstars.seti.org) sponsored by the NASA Astrobiology Institute and convened at the SETI Institute. PMID- 17407404 TI - High-resolution simulations of the final assembly of Earth-like planets. 2. Water delivery and planetary habitability. AB - The water content and habitability of terrestrial planets are determined during their final assembly, from perhaps 100 1,000-km "planetary embryos " and a swarm of billions of 1-10-km "planetesimals. " During this process, we assume that water-rich material is accreted by terrestrial planets via impacts of water-rich bodies that originate in the outer asteroid region. We present analysis of water delivery and planetary habitability in five high-resolution simulations containing about 10 times more particles than in previous simulations. These simulations formed 15 terrestrial planets from 0.4 to 2.6 Earth masses, including five planets in the habitable zone. Every planet from each simulation accreted at least the Earth's current water budget; most accreted several times that amount (assuming no impact depletion). Each planet accreted at least five water-rich embryos and planetesimals from the past 2.5 astronomical units; most accreted 10 20 water-rich bodies. We present a new model for water delivery to terrestrial planets in dynamically calm systems, with low-eccentricity or low-mass giant planets-such systems may be very common in the Galaxy. We suggest that water is accreted in comparable amounts from a few planetary embryos in a " hit or miss " way and from millions of planetesimals in a statistically robust process. Variations in water content are likely to be caused by fluctuations in the number of water-rich embryos accreted, as well as from systematic effects, such as planetary mass and location, and giant planet properties. PMID- 17407405 TI - M stars as targets for terrestrial exoplanet searches and biosignature detection. AB - The changing view of planets orbiting low mass stars, M stars, as potentially hospitable worlds for life and its remote detection was motivated by several factors, including the demonstration of viable atmospheres and oceans on tidally locked planets, normal incidence of dust disks, including debris disks, detection of planets with masses in the 5-20 M() range, and predictions of unusually strong spectral biosignatures. We present a critical discussion of M star properties that are relevant for the long- and short-term thermal, dynamical, geological, and environmental stability of conventional liquid water habitable zone (HZ) M star planets, and the advantages and disadvantages of M stars as targets in searches for terrestrial HZ planets using various detection techniques. Biological viability seems supported by unmatched very long-term stability conferred by tidal locking, small HZ size, an apparent short-fall of gas giant planet perturbers, immunity to large astrosphere compressions, and several other factors, assuming incidence and evolutionary rate of life benefit from lack of variability. Tectonic regulation of climate and dynamo generation of a protective magnetic field, especially for a planet in synchronous rotation, are important unresolved questions that must await improved geodynamic models, though they both probably impose constraints on the planet mass. M star HZ terrestrial planets must survive a number of early trials in order to enjoy their many Gyr of stability. Their formation may be jeopardized by an insufficient initial disk supply of solids, resulting in the formation of objects too small and/or dry for habitability. The small empirical gas giant fraction for M stars reduces the risk of formation suppression or orbit disruption from either migrating or nonmigrating giant planets, but effects of perturbations from lower mass planets in these systems are uncertain. During the first approximately 1 Gyr, atmospheric retention is at peril because of intense and frequent stellar flares and sporadic energetic particle events, and impact erosion, both enhanced, the former dramatically, for M star HZ semimajor axes. Loss of atmosphere by interactions with energetic particles is likely unless the planetary magnetic moment is sufficiently large. For the smallest stellar masses a period of high planetary surface temperature, while the parent star approaches the main sequence, must be endured. The formation and retention of a thick atmosphere and a strong magnetic field as buffers for a sufficiently massive planet emerge as prerequisites for an M star planet to enter a long period of stability with its habitability intact. However, the star will then be subjected to short-term fluctuations with consequences including frequent unpredictable variation in atmospheric chemistry and surficial radiation field. After a review of evidence concerning disks and planets associated with M stars, we evaluate M stars as targets for future HZ planet search programs. Strong advantages of M stars for most approaches to HZ detection are offset by their faintness, leading to severe constraints due to accessible sample size, stellar crowding (transits), or angular size of the HZ (direct imaging). Gravitational lensing is unlikely to detect HZ M star planets because the HZ size decreases with mass faster than the Einstein ring size to which the method is sensitive. M star Earth-twin planets are predicted to exhibit surprisingly strong bands of nitrous oxide, methyl chloride, and methane, and work on signatures for other climate categories is summarized. The rest of the paper is devoted to an examination of evidence and implications of the unusual radiation and particle environments for atmospheric chemistry and surface radiation doses, and is summarized in the Synopsis. We conclude that attempts at remote sensing of biosignatures and nonbiological markers from M star planets are important, not as tests of any quantitative theories or rational arguments, but instead because they offer an inspection of the residues from a Gyr-long biochemistry experiment in the presence of extreme environmental fluctuations. A detection or repeated nondetections could provide a unique opportunity to partially answer a fundamental and recurrent question about the relation between stability and complexity, one that is not addressed by remote detection from a planet orbiting a solar-like star, and can only be studied on Earth using restricted microbial systems in serial evolution experiments or in artificial life simulations. This proposal requires a planet that has retained its atmosphere and a water supply. The discussion given here suggests that observations of M star exoplanets can decide this latter question with only slight modifications to plans already in place for direct imaging terrestrial exoplanet missions. PMID- 17407406 TI - Coronal mass ejection (CME) activity of low mass M stars as an important factor for the habitability of terrestrial exoplanets. I. CME impact on expected magnetospheres of Earth-like exoplanets in close-in habitable zones. AB - Low mass M- and K-type stars are much more numerous in the solar neighborhood than solar-like G-type stars. Therefore, some of them may appear as interesting candidates for the target star lists of terrestrial exoplanet (i.e., planets with mass, radius, and internal parameters identical to Earth) search programs like Darwin (ESA) or the Terrestrial Planet Finder Coronagraph/Inferometer (NASA). The higher level of stellar activity of low mass M stars, as compared to solar-like G stars, as well as the closer orbital distances of their habitable zones (HZs), means that terrestrial-type exoplanets within HZs of these stars are more influenced by stellar activity than one would expect for a planet in an HZ of a solar-like star. Here we examine the influences of stellar coronal mass ejection (CME) activity on planetary environments and the role CMEs may play in the definition of habitability criterion for the terrestrial type exoplanets near M stars. We pay attention to the fact that exoplanets within HZs that are in close proximity to low mass M stars may become tidally locked, which, in turn, can result in relatively weak intrinsic planetary magnetic moments. Taking into account existing observational data and models that involve the Sun and related hypothetical parameters of extrasolar CMEs (density, velocity, size, and occurrence rate), we show that Earth-like exoplanets within close-in HZs should experience a continuous CME exposure over long periods of time. This fact, together with small magnetic moments of tidally locked exoplanets, may result in little or no magnetospheric protection of planetary atmospheres from a dense flow of CME plasma. Magnetospheric standoff distances of weakly magnetized Earth-like exoplanets at orbital distances 30 to < 40), and morbidly obese (BMI, >/= 40). MAIN OUTCOME MEASURES: Prevalence of obesity (compared with the age- and sex-matched adult Australian population); associations between obesity and morbid obesity in cardiac patients and adverse postoperative outcomes. RESULTS: 30.4% of patients had a BMI > 30 (28.6% obese, 1.8% morbidly obese) compared with an expected prevalence of 21.2%. Morbid obesity was associated with prolonged ventilation (adjusted odds ratio [OR], 2.4; 95% CI, 1.6-3.7), readmission to intensive care (adjusted OR, 2.2; 95% CI, 1.2-4.1), and length of stay > 14 days (adjusted OR, 2.1; 95% CI, 1.4-3.3). Both obesity and morbid obesity were associated with renal failure (adjusted ORs, 1.4 [95% CI, 1.1-1.7] and 2.9 [95% CI, 1.7-4.9], respectively) and deep sternal wound infection (adjusted ORs, 2.4 [95% CI, 1.5 3.8] and 7.2 [95% CI, 2.8-18.7], respectively). CONCLUSIONS: Obesity is 1.4 times more prevalent in patients having coronary artery bypass grafting or heart valve surgery in Victoria compared with the general adult Australian population. Both obesity and morbid obesity are associated with early morbidity, but not mortality, after operation. PMID- 17407432 TI - Australian general practice and pandemic influenza: models of clinical practice in an established pandemic. AB - To minimise the health impact of pandemic influenza, general practice will need to provide influenza-related and non-influenza primary health care, as well as contribute to the public health goal of disease control. Through interviews and workshops with general practitioners, nurses and policy leaders between March and July 2006, and literature analysis, we identified potential models of general practice in an established pandemic, and assessed their strengths and weaknesses. Three possible clinical models were identified: a default model of no change to service delivery; a streamed services model, where general practices reorganise themselves to take on either influenza-specific care or other clinical services; and a staff-determined mixed model, where staff move between different types of services. No single model or set of strategies meets the needs of all general practices to deliver and sustain the essential functions of primary health care during an established pandemic. Governments, general practice and the relevant peak professional bodies should decide before a pandemic on the suite of measures needed to support the models most suitable in their regions. Effective participation by general practice in a pandemic requires supplementary infrastructure support, changes to financial and staffing patterns, a review of legislation on medicolegal implications during an emergency, and intensive collaboration between general practices. PMID- 17407433 TI - Late-term abortion: what can be learned from Royal Women's Hospital v Medical Practitioners Board of Victoria? AB - In 2001, the Medical Practitioners Board of Victoria received a complaint from an Australian Government Senator regarding a late-term abortion carried out in February 2000 at the Royal Women's Hospital, Melbourne. Five years later, the complaint of professional misconduct was finally dismissed by the Board as being frivolous and vexatious. The action highlights a number of deficiencies in the way medical practitioner boards deal with complaints against medical practitioners; in particular, the Board's lack of discretion to deal with complaints lacking substance. Early mediation of the dispute between the Royal Women's Hospital and the Medical Practitioners Board could have avoided a great deal of suffering and expense. As a result of this case, it is likely that the Victorian Medical Practitioners Board will be given additional powers in the future to deal with complaints without merit. PMID- 17407434 TI - Maternal mortality and psychiatric morbidity in the perinatal period: challenges and opportunities for prevention in the Australian setting. AB - Maternal mortality associated with psychiatric illness in the perinatal period (pregnancy to the end of the first year postpartum) has until recently been under reported in Australia due to limitations in the scope of the data collection and methods of detection. The recent United Kingdom report Why mothers die 2000-2002 identified psychiatric illness as the leading cause of maternal death in the UK. Findings from the last three reports on maternal deaths in Australia (covering the period 1994-2002) suggest that maternal psychiatric illness is one of the leading causes of maternal death, with the majority of suicides occurring by violent means. Such findings strengthen the case for routine perinatal psychosocial screening programs, with clear referral guidelines and assertive perinatal treatment of significant maternal psychiatric morbidity. Data linkage studies are needed to measure the full extent of maternal mortality associated with psychiatric illness in Australia. PMID- 17407435 TI - Sezary syndrome and otitis externa. PMID- 17407436 TI - Putting results of a clinical trial into perspective. PMID- 17407437 TI - Accidental paracetamol poisoning. PMID- 17407438 TI - What may underlie recurrent purpura fulminans? AB - A woman presenting with recurrent purpura fulminans was eventually found to have inflammatory bowel disease. We suggest the inflammatory state resulted in a deficiency of functional protein C. PMID- 17407439 TI - Off-label use of medicines: consensus recommendations for evaluating appropriateness. PMID- 17407440 TI - Oocyte cryopreservation as an adjunct to the assisted reproductive technologies. PMID- 17407441 TI - Addressing the health costs of the Iraq war: the role of health organisations. PMID- 17407442 TI - Clinical paradigms revisited. PMID- 17407443 TI - The absence of many voices in protest. PMID- 17407444 TI - Snake-venom-derived Factor IX-binding protein specifically blocks the gamma carboxyglutamic acid-rich-domain-mediated membrane binding of human Factors IX and X. AB - A potent anticoagulant protein, IX-bp (Factor IX binding protein), has been isolated from the venom of Trimeresurus flavoviridis (habu snake) and is known to bind specifically to the Gla (gamma-carboxyglutamic acid-rich) domain of Factor IX. To evaluate the molecular basis for its anticoagulation activity, we assessed its interactions with various clotting factors. We found that the anticoagulation activity is primarily due to binding to the Gla domains of Factors IX and X, thus preventing these factors from recognizing phosphatidylserine on the plasma membrane. The present study suggests that ligands that bind to the Gla domains of Factors IX and X may have the potential to become novel anticoagulants. PMID- 17407445 TI - A structural insight into the inhibition of human and Leishmania donovani ornithine decarboxylases by 1-amino-oxy-3-aminopropane. AB - The critical role of polyamines in key processes such as cell growth, differentiation and macromolecular synthesis makes the enzymes involved in their synthesis potential targets in the treatment of certain types of cancer and parasitic diseases. Here we present a study on the inhibition of human and Leishmania donovani ODC (ornithine decarboxylase), the first committed enzyme in the polyamine biosynthesis pathway, by APA (1-amino-oxy-3-aminopropane). The present study shows APA to be a potent inhibitor of both human and L. donovani ODC with a K(i) value of around 1.0 nM. We also show that L. donovani ODC binds the substrate, the co-enzyme pyridoxal 5'-phosphate and the irreversible inhibitor alpha-difluoromethylornithine (a curative agent of West African sleeping sickness) with less affinity than human ODC. We have also determined the three-dimensional structure of human ODC in complex with APA, which revealed the mode of the inhibitor binding to the enzyme. In contrast with earlier reports, the structure showed no indication of oxime formation between APA and PLP (pyridoxal 5'-phosphate). Homology modelling suggests a similar mode of binding of APA to L. donovani ODC. A comparison of the ODC-APA-PLP structure with earlier ODC structures also shows that the protease-sensitive loop (residues 158-168) undergoes a large conformational change and covers the active site of the protein. The understanding of the structural mode of APA binding may constitute the basis for the development of more specific inhibitors of L. donovani ODC. PMID- 17407446 TI - Activation of NADPH oxidase by transforming growth factor-beta in hepatocytes mediates up-regulation of epidermal growth factor receptor ligands through a nuclear factor-kappaB-dependent mechanism. AB - The TGF-beta (transforming growth factor-beta) induces survival signals in foetal rat hepatocytes through transactivation of EGFR (epidermal growth factor receptor). The molecular mechanism is not completely understood, but both activation of the TACE (tumour necrosis factor alpha-converting enzyme)/ADAM17 (a disintegrin and metalloproteinase 17; one of the metalloproteases involved in shedding of the EGFR ligands) and up-regulation of TGF-alpha and HB-EGF (heparin binding epidermal growth factor-like growth factor) appear to be involved. In the present study, we have analysed the molecular mechanisms that mediate up regulation of the EGFR ligands by TGF-beta in foetal rat hepatocytes. The potential involvement of ROS (reactive oxygen species), an early signal induced by TGF-beta, and the existence of an amplification loop triggered by initial activation of the EGFR, have been studied. Results indicate that DPI (diphenyleneiodonium) and apocynin, two NOX (NADPH oxidase) inhibitors, and SB431542, an inhibitor of the TbetaR-I (TGF-beta receptor I), block up-regulation of EGFR ligands and Akt activation. Different members of the NOX family of genes are expressed in hepatocytes, included nox1, nox2 and nox4. TGF-beta up-regulates nox4 and increases the levels of Rac1 protein, a known regulator of both Nox1 and Nox2, in a TbetaR-I-dependent manner. TGF-beta mediates activation of the nuclear factor-kappaB pathway, which is inhibited by DPI and is required for up regulation of TGF-alpha and HB-EGF. In contrast, EGFR activation is not required for TGF-beta-induced up-regulation of those ligands. Considering previous work that has established the role of ROS in apoptosis induced by TGF-beta in hepatocytes, the results of the present study indicate that ROS might mediate both pro- and anti-apoptotic signals in TGF-beta-treated cells. PMID- 17407447 TI - Enzymatic cleavage specificity of the proalpha1(V) chain processing analysed by site-directed mutagenesis. AB - The proteolytic processing of procollagen V is complex and depends on the activity of several enzymes among which the BMP-1 (bone morphogenetic protein 1)/tolloid metalloproteinase and the furin-like proprotein convertases. Few of these processing interactions could have been predicted by analysing the presence of conserved consensus sequences in the proalpha1(V) chain. In the present study we opted for a cell approach that allows a straightforward identification of processing interactions. A construct encompassing the complete N-terminal end of the proalpha1(V) chain, referred to as Nalpha1, was recombinantly expressed to be used for enzymatic assays and for antibody production. Structural analysis showed that Nalpha1 is a monomer composed of a compact globule and an extended tail, which correspond respectively to the non-collagenous Nalpha1 subdomains, TSPN-1 (thrombospondin-1 N-terminal domain-like) and the variable region. Nalpha1 was efficiently cleaved by BMP-1 indicating that the triple helix is not required for enzyme activity. By mutating residues flanking the cleavage site, we showed that the aspartate residue at position P2' is essential for BMP-1 activity. BMP-1 activity at the C-terminal end of the procollagen V was assessed by generating a furin double mutant (R1584A/R1585A). We showed that, in absence of furin activity, BMP-1 is capable of processing the C-propeptide even though less efficiently than furin. Altogether, our results provide new relevant information on this complex and poorly understood mechanism of enzymatic processing in procollagen V function. PMID- 17407448 TI - The emergence of the mind: a borderline of human viability? PMID- 17407449 TI - Prognostic tests in babies: do they always help? PMID- 17407450 TI - Progress in predicting outcomes for extremely low birth weight infants: baby steps. PMID- 17407451 TI - New guidelines for newborn resuscitation. AB - The new guidelines from the International Liaison Committee on Resuscitation and American Heart Association/American Academy of Pediatrics for newborn resuscitation underline that efficient ventilation is the key to a successful resuscitation of the newly born infant. Compared with the former guidelines published in 1999, the major changes are (i) less emphasis on using supplemental oxygen when initiating resuscitation, (ii) no need for routine intrapartum oropharyngeal and nasopharyngeal suctioning for vigorous infants born to mothers with meconium staining of amniotic fluid, (iii) occlusive wrapping of very low birth weight infants <28 weeks to reduce heat loss is recommended, (iv) preference for the intravenous versus endotracheal route for adrenaline and (v) more emphasis on parental autonomy at the threshold of viability. A number of gaps in newborn resuscitation have been identified and discussed. CONCLUSION: The new guidelines for newborn resuscitation are more evidence-based than previously ones. However, still there is a need for further research and modifications. PMID- 17407452 TI - Understanding diagnostic tests 1: sensitivity, specificity and predictive values. AB - The usefulness of diagnostic tests, that is their ability to detect a person with disease or exclude a person without disease, is usually described by terms such as sensitivity, specificity, positive predictive value and negative predictive value. In this article, the first of the series, a simple, practical explanation of these concepts is provided and their use and misuse discussed. It is explained that while sensitivity and specificity are important measures of the diagnostic accuracy of a test, they are of no practical use when it comes to helping the clinician estimate the probability of disease in individual patients. Predictive values may be used to estimate probability of disease but both positive predictive value and negative predictive value vary according to disease prevalence. It would therefore be wrong for predictive values determined for one population to be applied to another population with a different prevalence of disease. CONCLUSION: Sensitivity and specificity are important measures of the diagnostic accuracy of a test but cannot be used to estimate the probability of disease in an individual patient. Positive and negative predictive values provide estimates of probability of disease but both parameters vary according to disease prevalence. PMID- 17407453 TI - Neurodevelopmental outcome in extremely low birth weight infants: what is the minimum age for reliable developmental prognosis? AB - AIM: We present a longitudinal study on the neurodevelopmental outcome in preterm infants with extremely low birth weight <1000 g (ELBW) to answer the question at which age a developmental prognosis can be given. METHODS: A group of 129 ELBW, median birth weight: 794 g (SD 123 g), gestational age: 27.0 weeks (SD 2.0 weeks), born between 1993 and 1998, were followed up to the age between 6 and 10 years (mean 8.5 years [SD 1.7 years]) and evaluated by neurodevelopmental and psychometric tests. The status of children without cerebral palsy was ranked into categories of major, minor and no developmental impairments. RESULTS: At the time of the last follow-up examination 17% of the children showed a major impairment including 9% cerebral palsy, 42% a minor impairment and 41% were normally developed. The longitudinal analysis of cases without cerebral palsy reveals that an assessment 'at term' can only give the correct developmental prognosis in 49% of the cases. At the corrected age of 12 months the prognosis is correct in 59% of the cases, whereas at the corrected age of 3 years 70% proves to be right. Diagnosis of cerebral palsy could be confirmed at the corrected age of 2 years with sufficient reliability. CONCLUSION: The neurodevelopmental evaluation of former preterm infants with a birth weight <1000 g demands a follow-up period of at least 6 years in order to make reliable statements. We are doubtful that follow-up testing completed prior to this age can yield reliable results. PMID- 17407454 TI - The psychological impact of genital anomalies on the parents of affected children. AB - BACKGROUND: There is scarce information on how parents cope with children with genital anomalies. Participants & methods: Twenty-six parents of 25 children with a median age of 0.5 years (r, 5 days-10.8 years) were recruited through the Scottish Genital Anomaly Network and had a quantitative assessment of parenting stress and coping patterns; a qualitative assessment by a semi-structured interview was also performed in19 parents. RESULTS: In five parents, the total stress score was above the 85th centile, denoting clinical levels of stress. Three parents showed reduced coping pattern scores for social support, self esteem and psychological stability and three showed a reduction in utilization of communication with medical professionals. The scores did not correlate with each other or the extent of genital anomaly in the child. Semi-structured interview analysis revealed parents' need for more knowledge about the imminent surgery, post-operative care and their desire for written information that could complement the time-restricted contact with the clinical team. CONCLUSION: In the majority of cases, parents did not display abnormal levels of stress or coping on quantitative assessment. The semi-structured interview provided further information about the parents' level of coping and potential for stress and highlighted the need for more effective exchange of clinical information at a critical period of the parent-child relationship. PMID- 17407455 TI - Ocular complications at the limits of viability. AB - AIM: To evaluate the incidence of retinopathy of prematurity (ROP) and other ocular morbidities in extremely premature infants. METHODS: A retrospective analysis of the prevalence and nature of ocular abnormalities in a cohort of 22 extremely pre-term infants born <25 + 0 weeks of estimated gestational age (GA) was performed. RESULTS: The children were grouped according to the observed disorder: 13 out of 22 (59%) neonates with mild ophthalmologic findings (ROP < or = stage II) [Group 1], 5 out of 22 (23%) infants with ROP stage III or more (Group 2) and 4 out of 22 (18%) neonates with severe ocular morbidity (congenital cataract, microphthalmia, partial optic nerve atrophy and corneal perforation due to an ulcer with lens protrusion), partly combined with ROP > or = stage III (three of four). One child of 22 (5%) needed laser therapy. Out of 22 admitted infants, 20 (91%) were discharged alive. CONCLUSION: The high rate of ocular morbidity besides ROP in extremely pre-term infants is noteworthy. Mechanisms influencing the postnatal development of the eye, especially their relation to the grade of prematurity and neonatological therapeutical strategies, require further investigations. PMID- 17407456 TI - Females born large for gestational age have a doubled risk of giving birth to large for gestational age infants. AB - AIM: To analyse if females born large for gestational age (LGA) have an increased risk to give birth to LGA infants and to study anthropometric characteristics in macrosomic infants of females born LGA. METHODS: The investigation was performed as an intergenerational retrospective study of women born between 1973 and 1983, who delivered their first infant between 1989 and 1999. Birth characteristics of 47,783 females, included in the Swedish Birth Register both as newborns and mothers were analysed. LGA was defined as >2 SD in either birth weight or length for gestational age. The infants were divided into three subgroups: born tall only, born heavy only and born both tall and heavy for gestational age. Multiple logistic and linear regression analyses were performed. RESULTS: Females, born LGA with regard to length or weight, had a two-fold (adjusted OR 1.96, 95% Cl 1.54-2.48) increased risk to give birth to an LGA infant. Females, born LGA concerning weight only, had a 2.6 (adjusted OR 2.63, 95%, 1.85-3.75) fold increased risk of having an LGA offspring heavy only and no elevated risk of giving birth to an offspring that was tall only, compared to females born not LGA. In addition, maternal obesity was associated with a 2.5 (adjusted OR 2.56, 95%, 2.20-2.98) fold increased risk of having an LGA newborn, compared to mothers with normal weight. CONCLUSION: Females, born LGA, have an increased risk to give birth to LGA infants, compared to mothers born not LGA. Maternal overweight increases this risk even further. PMID- 17407457 TI - Insulin-like growth factor 2 (IGF2) and IGF2 receptor gene variants are associated with fetal growth. AB - AIM: Normal variation in size at birth is a result of the interaction between fetal genetic factors and the maternal uterine environment. It is, however, unclear how genetic factors contribute to fetal growth. The insulin-like growth factor (IGF) system regulates uterine, placental and fetal development, thereby partially controlling the rate of fetal growth. The aim of this study was to investigate the associations between the neonatal birth weight and the genotypes of polymorphic loci in the IGF2 and IGF2 receptor (IGF2R) genes. METHODS: We determined the genotypes of two polymorphic loci in the IGF2 gene and four loci in the IGF2R gene in 884 pairs of normal Japanese mothers and their neonates, and compared the genotypes with the birth weight converted into standard deviation scores (SDSs) according to sex, parity and gestational weeks at delivery. RESULTS: There was a significant difference in birth weight SDSs among the three neonatal +3123/ApaI genotypes of the IGF2 gene; AA, AG and GG. There was also a significant difference in birth weight among the three neonatal c.901C > G genotypes of the IGF2R gene; CC, CG and GG. CONCLUSION: These findings indicate that both IGF2 and IGF2R gene variants are associated with fetal growth. PMID- 17407458 TI - The effect of the odour of mother's milk on breastfeeding behaviour of premature neonates. AB - AIM: To assess the effects of exposure to the odour of mother's milk on breastfeeding behaviour of premature neonates. METHODS: Thirteen preterm infants born at 30-33 weeks gestational age were tested. Seven infants were randomly assigned to the milk-odour condition, 6 to the water-control condition. During week 35 post-conceptual age, each baby was exposed to the appropriate odour stimulus for 120 sec. on 5 consecutive days immediately prior to a breastfeeding attempt. The breastfeeding bout following the final odour exposure session, and a second breastfeeding session shortly before the baby left the hospital, were analysed. Babies were weighed before and after each feeding session. RESULTS: During each breastfeeding session, babies in the milk-odour condition displayed longer sucking bouts and more bursts composed of >7 sucking movements, and also consumed more milk than the Control infants. The time spent in the hospital was significantly less for the milk-odour condition (median = 43 days vs. 55.5 days for Controls). CONCLUSION: Brief exposure to the odour of mother's milk prior to early breastfeeding trials had a positive effect on sucking behaviour and milk ingestion of preterm babies, which in turn resulted in a shortened stay in the hospital. PMID- 17407459 TI - Preliminary evidence of a sensitive period for olfactory learning by human newborns. AB - AIM: To test the hypothesis that a brief window of time immediately after delivery may be a particularly sensitive period for olfactory learning by human neonates. METHODS: Fifty-five vaginally delivered newborns were exposed to an odorant for 30 min beginning 4-37 min after birth (Early exposure) or 12-h post partum (Late exposure). Several days later, newborns' head orientation responses to the exposure odour versus an unfamiliar odour or an odourless control stimulus were tested. RESULTS: Infants in the Early exposure group spent significantly more time oriented towards the familiar scent rather than a novel odour (Z = 2.869; n = 28; p < 0.01), or an odourless stimulus (Z = 2.550; n = 28; p < 0.01). Infants in the Late exposure condition did not respond differentially to the exposure odour versus a novel odour (Z = 1.105; n = 27, p = 0.27), and spent more time oriented towards an odourless stimulus than to the exposure odour (Z = 2.042; n = 27, p < 0.05). CONCLUSION: Infants in the Early exposure group, but not in the Late exposure group, became familiar with the exposure odour and retained a memory trace of it during the test trials. PMID- 17407460 TI - Prenatal smoking exposure and psychiatric symptoms in adolescence. AB - AIM: Explore associations between smoking in pregnancy and psychiatric symptoms in the adolescent offspring. DESIGN/SUBJECTS: A prospective population based follow-up of 84 adolescents at 14 years of age, of whom 32 of the mothers reported smoking during pregnancy. MAIN OUTCOME MEASURES: The Achenbach System of Empirically Based Assessment (ASEBA), ADHD-Rating Scale IV, Autism Spectrum Screening Questionnaire (ASSQ), Children's Global Assessment Scale (CGAS), estimated IQ based on four subscales of WISC-III. RESULTS: Adolescents who were born by smokers had significantly more rule-breaking and aggressive behaviour, externalizing and total problems on the ASEBA than adolescents of non-smokers (p < 0.01), when reported by mothers, fathers and teachers. ADHD symptoms were reported more frequently (p < 0.05), and mothers also reported more internalizing symptoms (p < 0.05) and social problems (p < 0.001). The ASSQ sum score was higher (p < 0.001), and overall function as measured by the CGAS was lower (p < 0.01) for the smoking-exposed group. Associations were still present after controlling for possible confounding factors. CONCLUSION: Adolescents exposed to prenatal smoking had higher scores for both externalizing and internalizing psychiatric symptoms, which could not be explained by a broad range of possible psychosocial confounders. Thus, smoking in pregnancy may be a marker for increased risk of psychiatric symptoms in the offspring. PMID- 17407461 TI - The effects of smoking in pregnancy on factors influencing fetal growth. AB - AIM: To evaluate the influence of maternal smoking during pregnancy on factors influencing fetal growth. METHODS: Thirty newborns of smoking mothers were prospectively compared with 60 newborns of non-smoking mothers. Pre-albumin, albumin, triglycerides, glucose, insulin, insulin-like growth factor I, IGF binding protein 3, pH, lactic acid, erythropoietin and hemoglobin concentrations were measured in umbilical cord blood. RESULTS: Infants of smoking mothers had a significantly lower birth weight (3418 +/- 533 vs. 3863 +/- 503 g; p < 0.001), length (50.5 +/- 2,6 vs. 52.3 +/- 1.9 cm; p < 0.001) and head circumference (34.6 +/- 1.8 vs. 35.8 +/- 1.1 cm; p < 0.001) than controls. They also had significantly lower insulin (3.2 (2.0-4.9) vs. 5.8 (4.6-7.1) mU/L; p = 0.008), insulin-like growth factor I (54.4 +/- 32.5 vs. 93.8 +/- 54.5 microg/L; p = 0.001) and IGF binding protein 3 (1664 +/- 432 vs. 1943 +/- 421 microg/L; p = 0.01) concentrations, than controls. Infants of smoking mothers also had significantly higher hemoglobin (167 +/- 14 vs. 157 +/- 13 g/L; p = 0.002) and erythropoietin (42.3 (25.1-72.4) vs. 26.3 (21.9-30.9) U/L; p = 0.03) than controls, but not pH or lactate concentrations. There was no significant difference in pre-albumin, albumin, triglycerides and glucose concentrations. CONCLUSIONS: Smoking during pregnancy causes symmetrical fetal growth impairment, possibly due to decreased oxygen transport to the fetus and decreased concentrations of fetal insulin, insulin-like growth factor I and IGF binding protein 3. PMID- 17407462 TI - Breast milk levels of zinc and omega-6 polyunsaturated fatty acids and growth of healthy Chinese infants. AB - AIM: To examine the concentrations of zinc and omega-6 polyunsaturated fatty acids (omega-6 PUFAs) in breast milk, the impact of zinc on omega-6 PUFA metabolism, and the growth rate of infants. METHODS: Forty-one mother-term infant pairs from a rural area of northern Beijing, China, who were 1 month (n = 18, group I) and 3 months (n = 23, group II) old and exclusively breastfed, were studied. The dietary records and the concentrations of zinc and omega-6 PUFAs in the milk of lactating women and the increase in weight and length of their infants during 1 and 3 postnatal months were analysed. RESULTS: The dietary intakes of mothers in the two groups were the same, i.e. high in carbohydrate and low in fat, protein and energy. The maternal zinc intake was 7.5mg/d and thus reached only 34.6% of the current Recommended Nutrient Intake (RNI). The levels of zinc and arachidonic acid (AA, C20:4 omega-6) in the milk of group I were significantly higher than those in group II. Furthermore, significant positive correlations were found between the concentrations of zinc and AA in the breast milk and between the level of milk AA and weight gain. CONCLUSION: Zinc may be a co-factor and essential for essential fatty acids (EFA) metabolism. Thus suboptimal zinc intake may cause EFA imbalance. Further studies of Chinese rural mother-infant pairs are necessary to determine whether zinc supplementation should be recommended when lactation exceeds 3 months. PMID- 17407463 TI - Iron status at 1 and 6 years versus developmental scores at 6 years in a well nourished affluent population. AB - AIM: To examine the association between iron status at 1 and 6 years with development at 6 years. METHODS: In a longitudinal study of children (n = 77), iron status was measured at 1 and 6 years and the Icelandic Developmental Inventory, which evaluates children's motor and verbal development, was filled in by mothers near the children's sixth birthday. RESULTS: Children, iron-deficient at 1 year (n = 10), had lower fine motor development scores at 6 years than non iron-deficient (n = 56) (46.7 +/- 4.1 vs. 49.3 +/- 2.0; p = 0.011). Fine motor scores were also lower in children with depleted iron stores at 1 year (n = 26) than non-iron-depleted children (n = 40) (48.0 +/- 3.3 vs. 49.5 +/- 1.8; p = 0.045). Multiple regression analyses, with iron status indices at 6 years, showed that mean corpuscular volume along with male gender predicted significantly positively for expression (adj. R(2)= 0.15; p = 0.018; n = 73), while regression analyses, including iron status at 1 and 6 years, showed that haemoglobin at 6 years was positively associated with gross motor (adj. R(2)= 0.05; p = 0.038; n = 63). CONCLUSIONS: In an affluent society, iron deficiency and depleted iron stores at 1 year may contribute to worse fine motor developmental scores at 6 years, while low mean corpuscular volume and haemoglobin at 6 years might affect subsequent expression and gross motor scores negatively. PMID- 17407464 TI - Health and nutritional status of children of adolescent mothers: experience from a diarrhoeal disease hospital in Bangladesh. AB - AIM: The study aimed at assessing clinical and nutritional features and socioeconomic characteristics of the first birth-order children (1-48 months) of adolescent mothers. METHODS: Five hundred and thirty-nine first birth-order children of both sexes, aged 1-48 month(s) were studied. All study children had adolescent mothers aged < or =19 years (when attending hospital), who attended (as a patient) the Dhaka hospital of ICDDR, B during 2000-2005. A similar group of children (n = 540) of mothers aged 25-29 years (when attending hospital) constituted the comparison group. RESULTS: Malnutrition indicated by underweight [OR 2.3, 95% CI 1.7-3.1, p < 0.001], stunting [OR 2.1, 95% CI 1.5-2.8, p < 0.001], wasting [OR 1.8, 95% CI 1.3-2.7, p = 0.001], infancy (<12 months old) [OR 2.8, 95% CI 2.1-3.9, p < 0.001], duration of hospitalization (> or =48 h) [OR 1.6, 95% CI 1.2-2.2, p = 0.001], DPT immunization [OR 1.8, 95% CI 1.3-2.5, p = 0.001] and maternal illiteracy (no formal schooling) [OR 1.5, 95% CI 1.1-2.0, p = 0.007] were significantly associated with children of adolescent mothers, after adjusting for co-variates in the logistic regression analysis. Similar results were also observed when different indices of malnutrition (stunting, underweight or wasting) were added separately to the different models. CONCLUSION: Children of adolescent mothers are likely to be more malnourished, have lesser opportunities for DPT immunization and have longer duration of hospitalization. Adolescent mothers were also more likely to be illiterate. Therefore, the development of preventive and therapeutic strategies will be required to reduce morbidity and improve the health and nutrition status of both children and their adolescent mothers. PMID- 17407465 TI - Characteristics of infants admitted to hospital for persistent colic, and comparison with healthy infants. AB - AIM: To describe clinical characteristics of infants with colic admitted to hospital because of ongoing excessive crying (colic). METHODS: Characteristics of 104 infants admitted to hospital because of severe excessive crying (cases) were compared to those of 100 healthy thriving controls randomly selected from records of well baby clinics. RESULTS: Half of the cases were reported to cry excessively from the day of birth, and feeding changes had been recommended in 77%. A medical cause of excessive crying was identified in none of the cases. Almost all infants showed a rapid transition to normal crying behaviour during admission; none cried for >3 h per day. Mean gestational age and mean Apgar scores were slightly lower in cases than in controls. There was a trend of positive family history of atopy being more common in controls (45%) than in cases (38%, p = 0.14). Feeding problems were more common in cases (71%) than in controls (36%, 95% CI for difference 21.6%-46.9%). Pregnancy or birth complications were much more common in cases (85%) than in controls (37%; 95% CI 35%-58%). CONCLUSIONS: Infants with severe excessive crying show normalization of crying behaviour during hospital admission, and are unlikely to have medical causes for their colic. The most important risk factor for excessive crying was a complicated pregnancy or birth, suggesting that this might predispose parents to regard normal crying behaviour as excessive. PMID- 17407466 TI - Thyroid function in children with sepsis and septic shock. AB - AIM: A prospective study was conducted to determine thyroid hormone levels and their relationship to survival in children with septic shock and sepsis. METHODS: We estimated thyroid hormone levels (T3, T4, TSH, fT3 and fT4) in children with septic shock and compared with those in children with sepsis. RESULTS: Twenty four children (13 boys) with septic shock and 25 children (14 boys) with sepsis were enrolled. The median T3, T4, fT3, fT4 and TSH (95% confidence interval) were 40 (40-40.23) ng/dL, 4.45 (1.9-6.03) microg/dL, 1.85 (1.2-2.37) pg/mL, 0.77 (0.57 0.95) ng/dL, 0.51 (0.26-1.15) microIU/mL, respectively in children with septic shock group compared with 130 (98.28-163.48) ng/dL, 9.3 (7.66-10.63) microg/dL, 3.2 (3-4.27) pg/mL, 1.3 (1.1-1.4) ng/dL, 2.85 (1.07-3.61) microIU/mL, respectively, in children with sepsis. Children with septic shock who died (n = 12) had higher TSH levels compared to those who survived (p = 0.04). There was no difference in hormone levels between children with catecholamine responsive and catecholamine resistant septic shock. CONCLUSION: Children with septic shock had lower levels of T3, T4, fT3, fT4 and TSH compared to those with sepsis. Findings of our study suggest that derangement of thyroid functions in children is not an important factor contributing to the severity of septic shock. PMID- 17407467 TI - Five-years growth hormone (GH) treatment in adults with Prader-Willi syndrome. AB - AIM: We have previously shown that 1 year of growth hormone (GH) treatment to adults with Prader-Willi syndrome (PWS) has beneficial effects on body composition. The aim of the present observational study was to re-evaluate our cohort, with focus on long-term GH treatment. METHODS: Seven men and seven women, median age 31 years, were available for follow-up for 6 years. Nine were on GH treatment for 5 years. Body composition was measured with Dual Energy X-ray absorptiometry (DXA). RESULTS: In six GH treated patients with genetically verified PWS there was a substantial increase in lean body mass of 5 kg (p = 0.031) and a concomitant, however, non-significant, decrease in body fat of 5% (p = 0.156). The changes in the genetically verified patients without GH treatment were small and unsystematic. No compliance problems were reported. Only one non GH-treated woman developed overt diabetes. CONCLUSION: Despite inherent behavioural problems it was possible to continue GH injections for 5 years with sustained favourable effects on body composition without clinically, significant side effects. PMID- 17407468 TI - Age is of influence on midazolam requirements in a paediatric intensive care unit. AB - AIM: To test that age is of influence on midazolam requirements during prolonged mechanical ventilation in critically ill children. METHODS: Retrospective observational study of children (28 days-18 year) admitted between January 1st 2002 and January 1st 2005 who needed controlled mechanical ventilation for 5 days and initial sedation with midazolam were included. Exclusion criteria were psychomotor retardation, therapeutic use of midazolam, ventilator weaning within 5 days, kidney or liver failure. RESULTS: A total of 1186 children were admitted, of which 58 children were included. The children were divided into three age groups: 28 days-1 year (n = 28), 1-4 years (n = 16) and older than 4 years (n = 14). Within 2 days the children age 1-4 years received the maximum midazolam dosage (0.3 mg/kg/h). In addition, the mean total dose of midazolam was higher at all days for this age group. At day 5 none of the children between 1 and 4 years could be sedated with midazolam alone. CONCLUSIONS: Our data showed that children between 1 and 4 years needed higher doses of midazolam as compared to children who were younger and older. Furthermore, we observed that midazolam alone is a poor sedative for all age groups. The influence of and mechanisms for possible age related effects on midazolam requirements remain to be elucidated, as well as the position of midazolam as a first line drug for PICU sedation. PMID- 17407469 TI - Child behaviour after anaesthesia: association of socioeconomic factors and child behaviour checklist to the Post-Hospital Behaviour Questionnaire. AB - AIM: In the process of validation of the Swedish translation of the Post-Hospital Behaviour Questionnaire (PHBQ) to assess its relation to the Child Behaviour Checklist (CBCL) and to describe its relation to sociodemographic factors. METHOD: Three hundred and forty children 2-13 years studied in connection with elective procedures which included anaesthesia. Parents completed the CBCL forms before and after hospitalization as well as the PHBQ 2 weeks after hospitalization. RESULTS: In multivariate analysis of PHBQ total score independent risk factors were: age <5 (OR 2.4; CI 1.4-4.0), living in a one parent family (OR 4.4; CI: 1.6-12.6) and not living in a rural area (OR 1.6; CI: 1.0-2.6). The correlation between the total scores for PHBQ and CBCL for children aged 2-4 was moderate: (r = 0.38; p < 0.005). For children aged 4-7 there were significant correlations between PHBQ sleep anxiety and CBCL Other problems (r = 0.4; p < 0.01), PHBQ eating disturbances and CBCL total score (r = 0.3; p < 0.01), though none of these significant correlations were observed for children above 7 years of age. CONCLUSION: There is an association between PHBQ and CBCL which is weaker for older children. The reason for this might be that PHBQ is more sensitive in the younger age group and to minor changes in behaviour. Children younger than 5 years of age or living in a one parent family or not living in rural areas appear to have higher incidence of problematic behaviour in a 2 week follow up after anaesthesia. PMID- 17407470 TI - New reference values for the Alberta Infant Motor Scale need to be established. AB - AIM: The Alberta Infant Motor Scale (AIMS) is an infant developmental test, which can be used to evaluate motor performance from birth to independent walking. Between 1990 and 1992 Piper and Darrah determined reference values in a cohort in Canada. To our knowledge no study has been carried out to determine whether the Canadian data are representative for other countries. In the present study we aimed to establish whether the AIMS test needs new reference values for Dutch children. METHODS: Motor performance of 100 Dutch children, aged 0-12 months, was measured using the AIMS test. RESULTS: The mean percentile score of the Dutch children was 28.8 (+/-22.9, range 1-85). The percentile scores of the group were significantly lower than scores of the Canadian norm population (p < 0.001), whereby 75% of the Dutch children scored below the 50th percentile. These lower scores were not be explained by sex, racial differences or congenital disorders and were seen in all age groups. CONCLUSION: We conclude that new reference values on the AIMS test for the age group of 0-12 months need to be established for Dutch children. It is recommended that the need for new normative data is also determined in all other European countries. PMID- 17407471 TI - Parent-dependent barriers to varicella immunization in Israel: the importance of adequate information. AB - AIM: To examine the parent-dependent barriers to varicella immunization in Israel. METHODS: Data were collected by questionnaire from parents of children aged 1 to 18 years who attended an outpatient paediatric centre in five cities in Israel from March to May 2003. RESULTS: A total of 1474 parents completed the questionnaire. A history of chicken pox was reported in 850 children (57.8%), mostly (83%) between ages 1-6 years. Of the 624 children without a history of chicken pox, 213 (34.1%) were immunized against varicella. Immunization rates were significantly lower in families with lower parental education and in patients from cities with a lower socioeconomic ranking (p < 0.05). The main reasons for not being vaccinated (49%) were related to insufficient information about the vaccine itself and the vaccination process. These reasons were more common in families with lower parental education and from cities with lower socioeconomic ranking (p < 0.01). Other reasons included fear of adverse effects (12%) and waning immunity (6%), preference of natural illness over immunization (12%), and financial limitations (5%). CONCLUSIONS: Promoting parental knowledge about the varicella vaccine appears to be a key factor in improving the immunization rate, especially in families of lower education, and from cities with low socioeconomic ranking. PMID- 17407472 TI - A comparative study on bacterial cultures of urine samples obtained by clean-void technique versus urethral catheterization. AB - Contamination during urine collection causes difficulty in diagnosing infantile urinary tract infection (UTI). Though considered a gold-standard, suprapubic aspiration is traumatic and not always successful. Catheterization and clean void technique were often preferred but their relative usefulness has not been compared. OBJECTIVES: To compare the culture results of clean void urine (CVU) and catheter urine (CathU) from children below 2-years old known to have no UTI. We tested whether the false-positive rates of CVU were significantly different from that of CathU. METHODS: Paired CVU and CathU samples were collected from asymptomatic children admitted for micturiting cystourethrogram, and tested for leucocytes and nitrite, and bacterial culture using standard laboratory methods. RESULTS: Culture results for 98 children (82 boys, 16 girls; aged 6 +/- 4.3 months) were analysed. Analysis by presence/absence of growths showed good agreement between CVU and CathU for boys (Kappa 0.42) but poor for girls (Kappa 0.18). When analysed by colony counts, agreement was poor with CVU yielding more counts than CathU (Kappa 0.1 for boys and 0.04 for girls). If all mixed growth results were considered as contaminants, the false positive rates for CathU and CVU were similar whether the cut-off was defined as 10(3), 10(4) or 10(5)/mL. If mixed growth was believed to cause UTI, CathU produced less false-positive rates than CVU, though both rates were unacceptably high. CONCLUSION: In uncircumcized boys, both CVU and CathU were prone to contamination. Though the contaminating bacterial counts were lower in CathU culture, the false-positive rates were high with the lower cut-off CFUs. Contrary to previous recommendations, CathU should be interpreted similar to CVU to avoid false positive diagnosis of UTI. PMID- 17407473 TI - Intercountry adoptees in out-of-home care: a national cohort study. AB - AIM: To investigate risks of placements in out-of-home care for non-European adoptees. METHODS: Intercountry adoptees born outside Europe 1973-1984 (n = 16,522) were compared with same age peers from the majority population (n = 1,026,523) using national demographic and child welfare registers. Multivariate analyses were performed using logistic regression models, and odds ratios (OR) for different forms of out-of-home care placements were calculated. RESULTS: After adjustments for socio-demographic background variables, the OR:s for placements of intercountry adoptees in residential care from age 10 were 5.1 (95% CI 4.6-5.8) and 3.0 (95% CI 2.6-3.6) for placements in foster care from age 10. For placements in all forms of out-of home care up to age 10, the odds were on par with the majority population. Higher child age at adoption, origin from Latin America, single parent adoption and maternal age above 35 at birth of the child were identified as significant predictors of out-of-home care from age 10. CONCLUSION: Intercountry adoptees emerge as a risk group for placements in out-of home care during adolescence, especially for entries into residential care (in Sweden usually triggered by persistent behaviour problems). PMID- 17407474 TI - Childhood deaths due to electrocution in Adana, Turkey. AB - AIM: This retrospective study was conducted to determine the general features of childhood deaths due to electrocution, and draw the attention of community to these preventable deaths. METHODS: The autopsy records of the Morgue of the Adana Branch of the Turkish Forensic Medicine Council were used. Thirty-seven cases of childhood (under 18) electrocution during the period 1999-2004 were analyzed. RESULTS: Electrocution deaths comprised 0.8% of all the medico-legal autopsy cases (4515) during these 6 years. Thirty-one (83.8%) of the victims were male and only six (16.2%) were female. They were between 18 months and 18 years of age, and the mean age was 11.35, and all deaths were recorded as accidental. CONCLUSION: Medical units, and paramedical and educational centres should play important roles in formulating public health strategies to tackle preventable deaths such as electrocution. PMID- 17407475 TI - Medical problems in adolescents with myelomeningocele (MMC): an inventory of the Swedish MMC population born during 1986-1989. AB - AIM: To describe the prevalence of myelomeningocele (MMC) and the medical needs of adolescents, 15-18 years, with MMC in Sweden, at a time when they are on the threshold of adulthood, leaving paediatrics. METHODS: In a retrospective study, we identified all adolescents with MMC, born during 1986-1989 and living in Sweden on July 1, 2004. An inventory was agreed upon with questions concerning their medical problems and need for medical care. RESULTS: There were 175 persons 15-18 years of age, born with MMC or lipoMMC (prevalence 3.8 per 10,000). Hydrocephalus was seen in 86%, 31% had been operated because of tethered cord syndrome, and 6% for Chiari malformation symptoms. The majority had motor impairments. Clean intermittent catheterisation for bladder emptying was used by 85%, and 59% used enemas on a regular basis because of the neurogenic bowel dysfunction. Renal dysfunction was seen in 1.7% of the adolescents. CONCLUSION: Lifelong follow-up by many specialists, among others neurologists and neurosurgeons, urotherapists and urologists, orthopaedic surgeons and orthotists, is necessary for individuals with MMC. The complex medical situation, often in combination with cognitive difficulties, makes it necessary to coordinate medical services for this increasing group of adults with multiple impairments. PMID- 17407476 TI - Early cardiac involvement in children carrying the A3243G mtDNA mutation. AB - The phenotypic spectrum of the mitochondrial A3243G DKA mutation is highly variable, particularly when occuring in childhood. In contrast to the classical presentation in adulthood (MELAS syndrome; mitochondria! myopathy, encephalopathy, lactic acidosis and stroke-like episodes) children show a different pattern of symptoms, often without the typical encephalopathy or psychomotor regression. We present six children carrying the A3243G mtDNA mutation with a heteroplasmy above 50 % in muscle tissue. The age of diagnosis ranged from 2 weeks up to 14.5 years. The clinical presentation was rather non specific including muscle weakness, developmental delay and epilepsy. In this small pediatric group we detected presymptomatic cardiac involvement in five out of six children already at an early stage of disease. The cardiac pathology included cardiomyopathy and biventricular hypertrophy with rhythm disturbances (for example long QT-syndrome). The observed cardiac changes do not always increase the risk of cardiac deterioration; however, two of our patients died early on. CONCLUSION: We hypothesize that the A3243G mutation might be underdiagnosed, as patients could suffer from an unexplained cardiac death before the diagnosis is made. We advise performing regular repeated ECGs and echocardiography in all children carrying a A3243G mtDNA mutation independently from the presence of cardiac symptoms. PMID- 17407477 TI - Do wet diapers induce arousals in sleeping infants? AB - AIM: To find out whether simulated bladder voiding was able to induce arousals in sleeping infants. METHODS: Polygraphic recordings were performed in 34 infants and voiding was simulated by administering water into the diaper. RESULTS: Heart rate, respiratory frequency and electroencephalogram frequency did not change significantly during this procedure. Furthermore, simulated voiding was unable to cause an awakening or to induce body movements in sleeping infants. CONCLUSION: Simulated voiding was unable to induce arousals. PMID- 17407478 TI - Somatic and psychiatric co-morbidity in children with attention deficit hyperactivity disorder. AB - AIM: To describe the prevalence of somatic and psychiatric co-morbidity in children diagnosed with ADHD and other behavioural problems compared to this prevalence in children seen at the outpatient department without either of these conditions. METHODS: A retrospective controlled case study was conducted in 369 children. All children with ADHD were diagnosed by a clinical psychologist in a hospital setting according to the DSM IV classification. Co-morbidity was determined by pediatricians. RESULTS: Somatic co-morbidity was seen in 94 % of the children. However, there was no significant difference in the prevalence of somatic co-morbidity in patients with ADHD nor in patients with behavioural problems other than ADHD when compared with the control group. Only two differences slight were observed. In the ADHD group and the group with behavioural problems motor impairment was seen more often and in the control group constipation was diagnosed more frequently. CONCLUSION: Except for motor impairment, somatic co-morbidity of any kind does not seem to occur more frequently in children with ADHD. PMID- 17407479 TI - Investigation of a dizzy child. PMID- 17407480 TI - Infective endocarditis with hypocalcaemia and facial dymorphism in an adolescent girl. PMID- 17407481 TI - Successful therapy of paediatric ethylene glycol poisoning: a case report and annual survey by a regional poison centre. AB - Severe poisoning with ethylene glycol, often used as antifreeze, is a rare, life threatening event. Neurological symptoms are accompanied by metabolic acidosis with elevated anion gap and osmotic gap. We report on a 7-year-old boy suffering from severe ethylene glycol intoxication. Early diagnosis based on typical clinical signs and rapid initiation of specific therapy with ethanol resulted in complete and rapid recovery without haemodialysis becoming necessary. While one laboratory initially had not been able to detect ethylene glycol in the patient's blood, an ethylene glycol serum level of 3900 mg/L was measured by a second laboratory, the highest value ever reported in the scientific literature for paediatric cases. Ethylene glycol poisoning is verified by quantitative glycol analysis in serum, but only few laboratories are able to perform this investigation in emergency cases. Therefore, in the beginning diagnosis has to be based on patient's history and clinical signs. Every substantial suspicion of ethylene glycol poisoning has to be treated with an antidote (fomepizole or ethanol) immediately. CONCLUSION: Ethylene glycol poisoning is rare in the paediatric age group in our series of glycol poisonings. One should keep in mind, that early diagnosis and treatment due to clinical signs is important and haemodialysis usually is not necessary. PMID- 17407482 TI - Complete unilateral leg duplication with ipsilateral renal agenesis. AB - A case of a 5-month-old infant with complete duplication of the right leg and ipsilateral renal agenesis is presented. There was also a duplicated scrotum, posterior urethral valves with hydronephrosis, abnormal pelvic bones, and a ventricular septal defect. Complete caudal duplication is exceedingly rare. Associated defects are common. Several aetiological theories have been proposed. Among them, an underlying polytopic (acro-renal) developmental field defect has been suggested. It best explains the specific findings in our patient. However, the aetiology of caudal duplication syndromes may be multifactorial. In the light of twinning theories, associated duplication of hindgut derivates should be suspected in similar cases. Further research is needed. The motor-skeletal functional outcome after leg duplication surgery is mostly favourable. CONCLUSION: A case of complete unilateral leg duplication with ipsilateral renal agenesis is presented. It appears to support the theory of a polytopic developmental field defect. PMID- 17407483 TI - Haemolysis alters plasma flecainide levels in newborn infants. AB - Refractory neonatal supra-ventricular tachycardia may require the use of flecainide as anti-arrhythmic. Close control of plasma levels is mandatory due to pro-arrhythmic complications. However, inadvertent hemolysis during blood sampling may cause an increase in measured plasma flecainide concentrations. We conclude, therefore, that dosing of the drug should be always done with caution, and in case of suspected haemolysis plasma levels should be repeated with a new blood sample. PMID- 17407484 TI - Temporary henna tattoo is unsafe in atopic children. AB - Temporary henna tattoos have become increasingly popular as a safe alternative to permanent tattoos among American and European children and teenagers during the summer holidays. Currently, temporary henna tattoos contain not only henna, but also other additives such as para-phemylenediamine (PPD), which is considered to be the chemical agent that most frequently causes skin reactions associated with the use of commercial black henna. In this report, we describe an 11-year-old boy who applied a temporary black henna tattoo on his right arm during the summer holidays in Greece and developed a severe contact dermatitis at the tattoo site with residual hypopigmentation. He had no previous history of contact dermatitis, however he did suffer from seasonal allergic rhinitis and atopic dermatitis. Patch testing revealed a strong reaction to PPD, a substance commonly contained in temporary henna tattoo preparations. CONCLUSION: Henna tattoos are an increasing problem worldwide since they carry an increased risk of severe skin reactions; therefore we suggest that the use of temporary henna tattoos in children be discouraged. PMID- 17407485 TI - Mycoplasma respiratory tract infection complicated by Stevens-Johnson syndrome and surgical emphysema. PMID- 17407486 TI - Comments on 'Lead neurotoxicity in children: is prenatal exposure more important than postnatal exposure?'. PMID- 17407488 TI - Hypoglycaemia in an infant with severe pulmonary hypertension after prenatal exposure to nimesulide: coincidence or side effect? PMID- 17407489 TI - Is early identification of asymptomatic infants with 'mild' CFTR genotypes clinically useful? PMID- 17407491 TI - Ghrelin: a link between energy homeostasis and drug abuse? PMID- 17407492 TI - Ghrelin administration into tegmental areas stimulates locomotor activity and increases extracellular concentration of dopamine in the nucleus accumbens. AB - Ghrelin stimulates appetite, increases food intake and causes adiposity by mechanisms that include direct actions on the brain. Previously, we showed that intracerebroventricular administration of ghrelin has stimulatory and dopamine enhancing properties. These effects of ghrelin are mediated via central nicotine receptors, suggesting that ghrelin can activate the acetylcholine-dopamine reward link. This reward link consists of cholinergic input from the laterodorsal tegmental area (LDTg) to the mesolimbic dopamine system that originates in the ventral tegmental area (VTA) and projects to the nucleus accumbens. Given that growth hormone secretagogue receptors (GHSR-1A) are expressed in the VTA and LDTg, brain areas involved in reward, the present series of experiments were undertaken to examine the hypothesis that these regions may mediate the stimulatory and dopamine-enhancing effects of ghrelin, by means of locomotor activity and in vivo microdialysis in freely moving mice. We found that local administration of ghrelin into the VTA (1 microg in 1 microl) induced an increase in locomotor activity and in the extracellular concentration of accumbal dopamine. In addition, local administration of ghrelin into the LDTg (1 microg in 1 microl) caused a locomotor stimulation and an increase in the extracellular levels of accumbal dopamine. Taken together, this indicates that ghrelin might, via activation of GHSR-1A in the VTA and LDTg, stimulate the acetylcholine dopamine reward link, implicating that ghrelin is a part of the neurochemical overlap between the reward systems and those that regulate energy balance. PMID- 17407493 TI - Alcohol administration acutely inhibits ghrelin secretion in an experiment involving psychosocial stress. AB - The appetite-regulating hormones leptin and ghrelin are altered in alcoholism and influence the hypothalamic-pituitary-adrenal system. We investigated whether acute ethanol ingestion and stress exposure affect ghrelin secretion. Nine healthy male volunteers were exposed to a standardized laboratory stressor involving public speaking on 2 days. On the first day they ingested 0.6 g/kg ethanol and on the second a placebo drink 50 minutes before the stressor. Plasma ghrelin, cortisol, glucose, and insulin were measured at baseline and in eight subsequent samples obtained up to 120 minutes after drinking (75 minutes after stress onset). The stress test induced a transient and significant rise in cortisol, which was not altered by prior alcohol administration. No significant change of ghrelin, insulin or glucose levels was observed after the stressor. Ghrelin declined significantly within 15 minutes after alcohol drinking, fell to a minimum of 66% of baseline at 75 minutes and remained at that level until the last sample at 120 minutes. No significant ghrelin changes were observed during placebo experiments. Insulin and glucose were not significantly influenced by stress or by alcohol. We conclude that alcohol drinking acutely attenuates circulating ghrelin levels. This effect is more pronounced than would be expected from the calories ingested with alcohol, as compared with a prior report where liquid meals of different caloric content were administered. We could not observe a stress effect on ghrelin, which does not support a role for ghrelin in stress induced anorexia. PMID- 17407494 TI - Correlation between serum ghrelin levels and cocaine-seeking behaviour triggered by cocaine-associated conditioned stimuli in rats. AB - Ghrelin is a brain-gut peptide with growth hormone-releasing and appetite inducing activities. A growing body of evidence suggests that ghrelin may affect the central reward system and modulate the activity of the mesolimbic system. Recent clinical studies also showed a significant positive correlation between plasma ghrelin levels and craving in alcoholics. Accordingly, the present study investigated the potential role of serum ghrelin levels in the reinstatement of cocaine-seeking behaviour triggered by cocaine-associated cues. In addition, serum corticosterone levels were determined in the light of evidence suggesting that corticosterone plays a modulatory role in cocaine-seeking behaviour. Male Lister Hooded rats under a restricted diet regime were first trained to intravenously self-administer cocaine under a fixed ratio-1 schedule of reinforcement. Conditioned stimuli (CS: tone and cue-light on for 5 seconds) were presented contingently with cocaine delivery. Once a stable baseline of cocaine self-administration was observed, lever presses were extinguished to less than 30% of baseline rates by removing both cocaine and CS. Reinstatement of responding was then induced by re-exposure to cocaine-associated CS. Blood samples for the enzyme immunoassay determination of serum ghrelin and the radioimmunoassay determination of serum corticosterone levels were collected 30 minutes before the beginning of reinstatement sessions. Rats significantly reinstated their responding when exposed to CS. A positive and significant correlation was observed between ghrelin levels (r = 0.64; P < 0.05), but not corticosterone (r = 0.37; NS), and the increased active lever presses only in animals exposed to CS. These findings suggest a potential role of ghrelin in the modulation of cue-triggered reinstatement of cocaine-seeking behaviour. PMID- 17407495 TI - Region-specific down-regulation of Crhr1 gene expression in alcohol-preferring msP rats following ad lib access to alcohol. AB - Corticotropin-releasing hormone 1 receptors (CRH-R1) mediate increased behavioral sensitivity to stress and excessive alcohol self-administration following a history of dependence. It was recently demonstrated that the genetically selected alcohol-preferring msP rat line replicates many characteristics of the post dependent state, due to an innate up-regulation of the Crhr1 transcript in several limbic areas related to alcohol drinking motivation. Here, we examined whether voluntary alcohol consumption might be able to down-regulate Crhr1 transcript levels in msP rats in brain areas where elevated expression previously has been shown. Within central and medial amygdala (CeA, MeA), as well as the Nc. Accumbens, 2 weeks'ad lib access to alcohol led to a highly significant down regulation of the Crhr1 transcript. Alcohol-induced Crhr1 down-regulation was not seen in cingulate cortex. These data support that recruitment of CRH-R1 signaling within components of the extended amygdala drives excessive alcohol intake, and that alcohol is voluntarily consumed in part for its ability to reduce CRH-R1 activity in this region. PMID- 17407496 TI - Evidence for the role of dopamine D3 receptors in oral operant alcohol self administration and reinstatement of alcohol-seeking behavior in mice. AB - The present study examined the effects of the acute intraperitoneal (i.p.) administration of the selective dopamine (DA) D(3) receptor antagonist SB-277011A (10, 20 or 30 mg/kg i.p.) on the oral operant self-administration of alcohol in male C57BL/6N mice. These effects were compared with those of naltrexone (0.5, 1 and 2 mg/kg i.p.) and acamprosate (100, 200 and 400 mg/kg i.p.). Compared with vehicle, the acute administration of SB-277011A (10 or 20 mg/kg) did not significantly alter the operant self-administration of alcohol, whereas the 30 mg/kg dose significantly reduced alcohol intake (g/kg), the number of reinforcers, and the number of active lever presses. The oral self-administration of alcohol was not significantly altered by the acute administration of either naltrexone or acamprosate, compared with vehicle-treated mice. SB-277011A, naltrexone and acamprosate were also tested in a model of drug/cue-triggered reinstatement of alcohol-seeking behavior. In this model, neither naltrexone (2 mg/kg) nor acamprosate (400 mg/kg) prevented relapse to alcohol-seeking behavior. In contrast, SB-277011A significantly reduced reinstatement of alcohol seeking in a dose-dependent manner. Provided these results can be extrapolated to humans, they suggest that selective DA D(3) receptor antagonists may be useful in the pharmacotherapeutic management of alcohol intake and prevention of relapse to alcohol-seeking behavior. PMID- 17407497 TI - Difference in magnitude of psychostimulant-induced extracellular norepinephrine in the ventral tegmental area contributes to discrepant prefrontal dopamine outflow. AB - The dopamine (DA) efflux in the medial prefrontal cortex (mPFC) can be modulated by the interaction between afferent norepinephrine (NE) and somatodendritic DA in the ventral tegmental area (VTA). However, it is unclear how locally administered amphetamine (AMPH) or cocaine in the VTA results in discrepant response of DA efflux in the mPFC. In this study, intra-VTA infusion of AMPH (1000 microM) or cocaine (200 microM) in anesthetized rats was employed to study the concurrent profile of extracellular DA level in the VTA and mPFC. In addition, the extracellular NE levels during the intra-VTA infusion of these two psychostimulants were analyzed to compare their effects on prefrontal DA efflux. During the intra-VTA infusion of AMPH, both extracellular DA and NE increased significantly in the VTA (270 +/- 12% and 819 +/- 40%, respectively). Meanwhile, the DA efflux in the mPFC elevated significantly. During the intra-VTA infusion of cocaine, the extracellular DA and NE in the VTA also increased (271 +/- 21% and 150 +/- 15%, respectively). However, the DA efflux decreased significantly in the mPFC. Noteworthy, the increase of extracellular NE in the VTA was much more robust via AMPH infusion, as compared with cocaine. It is suggested that AMPH and cocaine enhance the extracellular NE concentrations in the VTA in different magnitudes, which in turn contribute to discrepant profiles of distal DA efflux in the mPFC. PMID- 17407498 TI - Differential effects of ethanol on IFN-gamma- and TNF-alpha-producing splenic T lymphocytes in a murine model of gram-negative pneumonia. AB - The incidence of bacterial pneumonia is increased in alcoholic patients. Alcohol consumption has been shown to impair cytokine production. Tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) are critical for host defense against Klebsiella pneumoniae (K. pneumoniae). In order to examine the influence of alcohol on the immune response to infection, we investigated the frequency of TNF-alpha and IFN-gamma produced by splenic T-lymphocytes in a murine model of gram-negative pneumonia initiated after 8 days of alcohol treatment. Thirty-two Balb/c mice were pretreated with ethanol (3 mg/g body weight) or saline intraperitoneally over 8 days. On day 7 half of each group was administered K. pneumoniae. Mice were sacrificed 24 hours later to excise lungs and liver for histological assessment and spleens for cell isolation. IFN-gamma- and TNF-alpha producing CD4(+) and CD8(+) lymphocytes were determined by FACS analysis. In mice with Klebsiella infection, the percentages of IFN-gamma-producing CD4(+) (P < 0.01) and CD8(+) (P < 0.01) were significantly decreased, the percentages of TNF alpha-producing CD4(+) (P = 0.01) and CD8(+) (P < 0.01) T cells were significantly elevated after alcohol treatment compared with mice with saline treatment. The histological assessment showed an aggravation of K. pneumoniae induced pneumonia in alcohol-treated mice. Alcohol differentially affects IFN gamma and TNF-alpha production in Klebsiella-infected mice. Both effects obviously led to a weakened immune response as seen by increased histological damage. This suggests a role of T cells in the increased susceptibility of the alcoholic host to nosocomial infection due to inadequate cytokine response. PMID- 17407499 TI - The effects of p-chloroamphetamine, methamphetamine and 3,4 methylenedioxymethamphetamine (ecstasy) on the gene expression of cytoskeletal proteins in the rat brain. AB - Repeated administration of beta-phenylalkylamines is known to produce neuronal changes in the central and peripheral nervous systems of mammals. It is suggested that various components of the cytoskeleton undergo profound alterations after amphetamine use and misuse, contributing to behavioral changes and neurotoxicity. Here we studied the expression of microtubule-associated protein 2 (MAP2) and beta-actin after repeated intraperitoneal applications with equimolar doses of p chloroamphetamine (PCA), methamphetamine (METH) and 3,4 methylenedioxymethamphetamine (MDMA) in the brain of male Wistar rats. Effective (molecular) pharmacological doses (ED) were derived and used for the calculation of (molecular) pharmacological indices (PI). Besides clear but different dose response curves on the toxicity of the drugs, in situ hybridization and Western blot analysis revealed that repeated administration of these compounds resulted in different substance- and dose-dependent changes in MAP2 gene expression, e.g. in the frontoparietal somatosensoric cortex. In contrast, the expression of beta actin was not influenced by any of the compounds at the dose levels tested. Lethal doses were determined with 2.1 (PCA), >5.1 (METH) and 8.4 mg/kg/day (MDMA). Linear and non-linear repeat-dose lethality was observed for MDMA and PCA, respectively, whereas METH was non-lethal in the dose range used. Values for ED(MAP2) were 0.3, 0.52 and >16.8 mg/kg/day, and therefore those for PI(MAP2) were 20, 4, and 0.5 for METH, PCA and MDMA, respectively. Although the results on mortality did not reflect changes in MAP2 gene expression, they suggest a remarkable difference for those amphetamines without substituents or with a halogen atom at the paraposition of the benzene ring, such as METH or PCA, when compared with MDMA-like substances. PMID- 17407500 TI - Phosphatidylethanol as a sensitive and specific biomarker: comparison with gamma glutamyl transpeptidase, mean corpuscular volume and carbohydrate-deficient transferrin. AB - Phosphatidylethanol (PEth), a direct ethanol metabolite, is detectable in blood for more than 2 weeks after sustained ethanol intake. Our aim was to assess the usefulness of PEth [comparing sensitivity, specificity and the area under the curve (AUC)] as compared with carbohydrate-deficient transferrin (CDT), gamma glutamyl transpeptidase (GGT) and mean corpuscular volume (MCV), calculating the results from sober patients against those from alcohol-dependent patients during withdrawal. Fifty-six alcohol-dependent patients (ICD-10 F 10.25) in detoxification, age 43 years, GGT 81 U/l, MCV 96.4 fl, %CDT 4.2, 1400 g ethanol intake in the last 7 days (median), were included in the study. Over the time of 1 year, 52 samples from 35 sober forensic psychiatric addicted in-patients [age 34 years, GGT 16 U/l, MCV 91 fl, CDT 0.5 (median)] in a closed ward were drawn and used for comparison . PEth was measured in heparinized whole blood with a high-performance liquid chromatography method. GGT, MCV and %CDT were measured using routine methods. A receiver operating characteristic curve analysis was carried out, with 'current drinking status' (sober/drinking) as the state variable and PEth, MCV, GGT and CDT as test variables. The resulting AUC was 0.974 (P < 0.0001, confidence interval 0.932-1.016) for PEth. At a cut-off of 0.36 micromol/l, the sensitivity was 94.5% and specificity 100%. The AUC for CDT, GGT and MCV were 0.931, 0.894 and 0.883, respectively. A significant Spearman's rank correlation was found between PEth and GGT (r = 0.739), CDT (r = 0.643), MVC (r = 0.639) and grams of ethanol consumed in the last 7 days (r = 0.802). Our data suggest that PEth has potential to be a sensitive and specific biomarker, having been found in previous studies to indicate longer lasting intake of higher amounts of alcohol. PMID- 17407501 TI - The validity of the laboratory marker combinations DOVER and QUVER to detect physician's diagnosis of at-risk drinking. AB - Especially in situations where it might be favorable for the patient to dissimulate the existing alcohol problem, 'objective' laboratory tests can be helpful. In this study we report validation of the two combinations DOVER (DOctor VERified) and QUVER (QUestionnarie VERified) of the biological markers percent carbohydrate-deficient transferrin (%CDT) and gamma-glutamyl-transferase (gamma GT) to detect patients that have been identified by their physicians with at-risk drinking behavior. Fifty-eight general practitioners (GPs) participated at two study sites in South-West Germany. Patients filled in a questionnaire that included the alcohol use disorders identification test (AUDIT) and gave a blood sample. The GP recorded his/her assessment about the presence of an alcohol related disorder in the patient. Receiver operating characteristics (ROC) analyses of the marker combinations DOVER and QUVER were performed. A total of 2940 patients participated in the study, of which 2496 completed data sets that could be used for further analysis. The area under the curve (AUC) of 79.5% for DOVER and 77.2% (QUVER) are in a higher range than the values for gamma%CDT (75.7%) or gamma-GT (72.5%) and %CDT (64.5%) and suggest superiority of the proposed marker combinations. Cross-validation results were almost identical with 76.6% and 73.3% for DOVER and QUVER, respectively. Our analysis demonstrated that the combination of the markers gamma-GT and %CDT with the physician's judgement of the condition as reference was superior to the use of single markers. PMID- 17407502 TI - Association between the BclI glucocorticoid receptor polymorphism and smoking in a sample of patients with obstructive airway disease. AB - Glucocorticoids seem to mediate the effect of stimulant drugs such as nicotine. Several studies have pointed to an association between the BclI polymorphism in the glucocorticoid receptor gene and increased glucocorticoid effects. We analysed the association of smoking behaviour and the BclI polymorphism using a case-control design within the framework of a larger pharmacogenetic study. A total of 327 Caucasian patients with asthma or chronic obstructive pulmonary disease from 39 German general practices gave informed consent to take part in the study. They filled in questionnaires concerning their smoking behaviour and were genotyped for the BclI polymorphism. The genotype frequencies for non smokers (n = 251; CC, 0.42; CG, 0.46; GG, 0.12) as well as for smokers (n = 76; CC, 0.29; CG, 0.55; GG, 0.16) were consistent with the Hardy-Weinberg equilibrium. The proportion of smokers was significantly lower among carriers of the CC-genotype (22/127 = 17%) compared with carriers of the G-allele (54/200 = 27%; chi2 = 4.08; P = 0.04). Within the group of smokers, the proportion of heavy smokers (> 19 cigarettes/day; median) was reduced in C-homozygous patients when compared with carriers of the G-allele (7/22 = 32% versus 31/54 = 57%; chi2 = 4.09; P = 0.04). Stepwise logistic regression analysis also pointed to an association between the CC-genotype and a reduced probability of being a smoker (odds ratio = 0.55; 95% confidence interval = 0.30-1.00; P = 0.05) controlling for other predictors. In summary, this study provides evidence that the BclI polymorphism might play a role in the maintenance and severity of nicotine dependence. PMID- 17407503 TI - Significant impact of MTHFR C677T polymorphism on plasma homovanillic acid (HVA) levels among alcohol-dependent patients. AB - The enzyme 5,10-methylenetetrahydrofolate reductase (MTHFR) synthesizes 5 methyltetrahydrofolate. It plays a critical role in homocysteine metabolism. A high impact of MTHFR C677T polymorphism on plasma homocysteine levels has been observed among alcoholics. Recent studies indicate that homocysteine has toxic effects on dopaminergic neurons. Thus it lowers levels of homovanillic acid (HVA) in the striatal region in rats. Alcoholics had significantly lower plasma HVA concentrations compared with healthy controls. Aim of this study is to elucidate whether HVA plasma levels in alcoholics are influenced by MTHFR C677T polymorphism. A total of 142 alcohol-dependent patients and 101 healthy controls were examined regarding plasma HVA concentration and MTHFR C677T genotype. Blood samples of alcoholics were obtained after a minimum of 22 days of abstinence. Among alcohol-dependent patients MTHFR C677T polymorphism was significantly associated with plasma HVA levels: carriers of MTHFR C677T T-allele had significantly lower HVA plasma levels compared with homozygote carriers of C allele: 11.9 ng/ml versus 14.4 ng/ml (chi2: 5.39; P = 0.02). In healthy control subjects plasma HVA levels did not differ significantly between MTHFR C677T T allele carriers and homozygote carriers of C-allele: 15.1 ng/ml versus 15.3 ng/ml (chi2: 0.04; P = 0.82). The data suggest an influence of MTHFR C677T polymorphism on plasma HVA among alcohol-dependent patients. This might be due to neurotoxic effects of homocysteine on the dopaminergic system or direct impairment of monoamine metabolism. Future studies should try to elucidate whether this effect is reversible during alcohol abstinence. PMID- 17407504 TI - Dopamine receptor genes (DRD2, DRD3 and DRD4) and gene-gene interactions associated with smoking-related behaviors. AB - Cigarette smoking, like many addictive behaviors, has a genetic component, and the dopamine D2-like receptor genes (DRD2, DRD3 and DRD4) are candidates for contributing to these behaviors. Phenotypic information concerning smoking related behaviors from a nationally representative sample of research volunteers was analyzed for association with polymorphisms in these genes. Genotype status at the DRD2 intron 2 simple tandem repeat was related to cigarettes per day (P = 0.035) and heaviness of smoking index (P = 0.049). The presence of the glycine allele at the S9G polymorphism of the DRD3 gene was associated with frequency/quantity measures of smoking [log-transformed time to first cigarette (P = 0.031) and heaviness of smoking index (P = 0.035)]. There was a trend for DRD4 long alleles of the variable number of tandem repeats polymorphism to be associated with reduced severity of three withdrawal symptoms [desire/craving (P = 0.054); anger/irritability (P = 0.10); and trouble sleeping (P = 0.068)]. Interactions between genotypes at all three genes were associated with nervousness (P = 0.020) and trouble sleeping (P = 0.015). An interaction between DRD2 and DRD3 was found for trouble concentrating (P = 0.020). These relationships present possible dopamine-related responses to nicotine that warrant further study. PMID- 17407505 TI - Reduced fMRI activation of an occipital area in recently detoxified alcohol dependent patients in a visual and acoustic stimulation paradigm. AB - Chronic alcohol consumption is associated with neural damage that manifests in deficits in information processing. Previous studies evaluated higher cognitive functions such as working memory, but basic sensory information processing circuits have never been investigated before. Therefore, we applied a simple visual and acoustic stimulation paradigm in this functional magnetic resonance imaging (fMRI) pilot study. Nine recently detoxified male alcohol-dependent patients and nine healthy volunteers were presented a well-established 6-Hz checkerboard and auditory stimuli in the form of drumbeats in a block-design fMRI paradigm. During visual and acoustic stimulation, alcoholics and controls activated widespread occipital and temporal brain areas, as well as parts of the dorsolateral prefrontal cortex and thalamus. In a comparison of the stimulation induced activation of alcoholics and controls, the alcoholics showed a significantly lower blood oxygen level dependent (BOLD) signal in an extended bilateral occipital area (P < 0.001) as compared with healthy controls. In no region was the BOLD signal significantly higher in the alcohol-dependent subjects compared with controls. The reason for the new finding of a highly significant lower activation of the occipital cortex is unclear. It is in line with studies of neuropsychological tests in recently detoxified alcohol-dependent patients that also reported deficits in visual abilities. Attention deficits or a persisting neuronal alteration in the first weeks of alcohol abstinence may have contributed to this result. PMID- 17407506 TI - Gray matter volume abnormalities and externalizing symptoms in subjects at high risk for alcohol dependence. AB - Reduced right amygdala volumes have been reported in young, alcohol-naive subjects at high risk (HR) for alcohol dependence. The differences in brain morphometry have been associated with an excess of externalizing behaviors in these subjects. This may reflect a neurobiological vulnerability to alcohol dependence. Existing Magnetic Resonance Imaging (MRI) studies on these subjects have examined only a few, pre-selected brain regions using the manual regions of interest (ROI) approach. MRI of HR subjects (n = 20) and age, sex, and handedness matched low-risk (LR) subjects (n = 21) were analyzed using optimized voxel-based morphometry and ROI approach. The externalizing symptoms of these subjects and their fathers were measured using the Semi-Structured Assessment for the Genetics of Alcoholism. HR subjects had significantly smaller volumes of superior frontal, cingulate and parahippocampal gyri, amygdala, thalamus and cerebellum. These gray matter volumes correlated negatively with externalizing symptoms scores. Subjects at HR for alcoholism have reduced volumes of critical areas of brain gray matter, which are associated with increased externalizing symptoms. These represent key endophenotypes of alcoholism. PMID- 17407508 TI - Laparoscopic urological training programmes: the need for a consensus on minimum standards. PMID- 17407509 TI - Day-case urological surgery: are we improving? PMID- 17407510 TI - What activities are safe with kidney stones? A review of occupational and travel advice in the UK. PMID- 17407511 TI - Overactive bladder syndrome in older people. AB - The overactive bladder symptom complex (OAB) is the commonest cause of urinary incontinence in older people, and is usually due to underlying detrusor overactivity, and as such is a treatable condition. Older people are a heterogeneous group, which includes fit community-dwelling individuals and those with significant medical comorbidity; thus the requirements of care for this group are many and varied. The International Continence Society definition of the frail elderly, those aged >65 years with continence problems, who by virtue of comorbidity are house-bound or living in an institution, is clearly not applicable to all. However, many conditions begin to appear in later life and practitioners need to be aware of the need to manage these, and their treatment, when dealing with older people. Studies of medication for OAB have included the elderly and there is evidence of an equivalent benefit in younger people. The impact of treatment on the cognitively impaired and those receiving acetylcholinesterase inhibitors is discussed. PMID- 17407512 TI - Evaluation and significance of circulating epithelial cells in patients with hormone-refractory prostate cancer. AB - OBJECTIVE: To determine the feasibility of using flow cytometry fluorescence activated cell sorting (FACS) analysis for detecting circulating epithelial cells (CECs) in patients with hormone-refractory prostate cancer (HRPC), and to determine whether CECs can be used to predict survival in these patients. PATIENTS AND METHODS: Several prognostic models that include routinely used clinical and laboratory variables for predicting survival in men with HRPC have been reported; the presence of CECs measured by reverse transcriptase-polymerase chain reaction for prostate-specific antigen (PSA) in patients with HRPC is an independent prognostic factor for survival. CECs detected by FACS analysis correlate with advanced stage and poor survival outcome. A retrospective study was conducted to assess the presence of CECs by FACS analysis in metastatic HRPC patients initiating systemic chemotherapy with a taxane-based regimen. The association between clinical variables previously described and the presence of CECs along with the effect of the magnitude of CECs on survival was calculated, in 41 patients with HRPC, all of whom had peripheral blood collected for FACS analysis. RESULTS: Except for four patients, all those with metastatic HRPC had detectable CECs. Among these patients, the number of CECs/mL was correlated with age, serum PSA level and serum alkaline phosphatase (ALP). Higher serum levels of PSA and ALP predicted a poor survival outcome. Similarly, patients with < or =1.8 CECs/mL had a significantly longer survival than those with more CECs/mL (P = 0.02). With a median follow-up of 15.4 months, the median overall survival for all patients was 18.4 months. CONCLUSIONS: The presence of more CECs in patients with metastatic HRPC was associated with a poorer survival outcome; levels of > or =1.8 CECs/mL were associated with a shorter survival in patients with metastatic HRPC. PMID- 17407513 TI - Differences and commonalities in the management of locally advanced prostate cancer: results from a survey of oncologists and urologists in the UK. AB - OBJECTIVE: To determine the management practices used by UK oncologists and urologists for patients with locally advanced (non-metastatic) prostate cancer. METHODS: Using a postal questionnaire, 155 practising specialist oncologists and urologists were surveyed in the UK. Their views were sought on a multidisciplinary approach to the management of locally advanced prostate cancer and their current management practices. RESULTS: Over half of respondents recognized the need for both oncologists and urologists to take the lead in management decisions, but almost as many still expected the sole responsibility to lie within their own speciality. Radical radiotherapy (RT) was considered the current optimum treatment by most respondents, but 22% of urologists thought that radical prostatectomy is optimal. Most responders would use luteinizing hormone releasing hormone agonists as neoadjuvant and adjuvant to RT but there was significant variation in the favoured duration of treatment of these drugs, and in the dose of RT. CONCLUSION: This survey suggests that there are still wide variations in the management practices for locally advanced prostate cancer in the UK, and between urologists and oncologists. Improved consensus guidelines are required. PMID- 17407514 TI - Characteristics of incidental prostatic adenocarcinoma in contemporary radical cystoprostatectomy specimens. AB - OBJECTIVES: To investigate the relationship between prostate-specific antigen (PSA) level and tumour volume for incidental adenocarcinoma of the prostate found in cystoprostatectomy (CP) specimens, and to analyse the incidence of clinically significant prostate cancers in CP specimens and the biochemical recurrence of incidental prostate cancers on short-term follow up. PATIENTS AND METHODS: Complete data from 97 of 105 prostates from CP specimens were available. Prostates were thoroughly analysed and sectioned at 2 mm intervals. PSA levels and the findings at digital rectal examination before surgery were obtained prospectively. None of the patients had any evidence of prostate cancer before CP. RESULTS: Incidental prostate cancer was detected in 58 of 97 (60%) of the CP specimens; of these, 31 (53%) were significant according to the definition of Stamey et al. There was a weak correlation between tumour volume and PSA level, weighted solely by the four larger-volume cancers. The median PSA level for patients with and without prostate cancer was not significantly different (3.1 vs 1.1 ng/mL, P = 0.06). The follow-up of the 35 patients alive with prostate cancer showed four PSA recurrences (PSA >0.02 ng/mL) with one distant metastasis after a median follow-up of 3 years. None of the patients with insignificant tumours developed biochemical recurrence. CONCLUSIONS: The weak correlation between PSA level and tumour volume in these patients supports the argument that PSA is largely produced by benign prostatic hyperplasia and is therefore a poor screening tool for asymptomatic healthy men. Most incidental prostate cancers in CP specimens are significant, contrary to previous analyses, but have little practical importance in terms of oncological outcome. PMID- 17407515 TI - Small renal masses: the value of contrast-enhanced colour Doppler imaging. AB - OBJECTIVE: To assess the value of a microbubble-based ultrasonographic contrast agent for enhancing blood vessels in colour Doppler imaging (CDI) of small renal masses. PATIENTS AND METHODS: Fifty-one patients with small renal masses (< 3 cm in diameter) had prospective CDI before and after intravenous administration of the contrast agent Levovist (Schering, Berlin, Germany). The degree of tumour vascularity was subjectively graded from 0 to IV (indicating an increasing vessel count). Furthermore, peak systolic velocity (PSV), resistive index, and pulsatility index were measured. The CDI findings were then compared with those obtained at histopathological examination. RESULTS: Intra- and/or peritumoral vessels were detected in 26 lesions (51%) by unenhanced CDI and in 48 by enhanced CDI (94%; P = 0.006, McNemar test). Higher grades of tumour vascularity (grade III and IV) were more common in malignant renal masses (P < 0.01). There were PSVs of >80 cm/s only in malignant lesions. Based on receiver operating characteristic analysis, enhanced CDI (area under the curve 0.789) was more accurate than unenhanced CDI (0.576) for differentiating benign from malignant renal masses (P < 0.004). CONCLUSION: Enhanced CDI is better than unenhanced CDI for detecting tumour vascularity, and for discriminating between benign and malignant small renal masses. PMID- 17407516 TI - Blood loss and postoperative complications associated with transurethral resection of the prostate after pretreatment with dutasteride. AB - OBJECTIVE: To determine whether pretreatment with dutasteride, a dual 5alpha reductase inhibitor (5ARI), reduces surgical blood loss or postoperative complications in patients with benign prostatic hyperplasia (BPH) who undergo transurethral resection of the prostate (TURP). PATIENTS AND METHODS: This double blind, randomized, placebo-controlled, multicentre study comprised 214 patients with BPH. Placebo was compared with dutasteride 0.5 mg/day 2 weeks before and after TURP, or 4 weeks before and 2 weeks after TURP. Surgical blood loss was measured using a haemoglobin photometer (HemoCue AB, Angelholm, Sweden) and postoperative adverse events were recorded. Microvessel density (MVD) was calculated by immunostaining and light microscopy of the prostatic chips. RESULTS: Although dutasteride reduced serum dihydrotestosterone (DHT) by 86-89% in 2-4 weeks, and intraprostatic DHT was approximately 10 times lower than in the placebo group, the (adjusted) mean haemoglobin (Hb) loss during surgery was 2.15 2.55 g Hb/g resectate with no significant difference in blood loss between the groups either during or after TURP. Clot retention occurred in 6-11% and urinary incontinence in 14-15% of patients during the 14 weeks after TURP, with no difference between the groups. The MVD at TURP was also similar for all groups. CONCLUSION: There were no significant reductions in blood loss during or after TURP or complications afterward with dutasteride compared with placebo, despite significant suppression of intraprostatic DHT. Blood loss and transfusion rates in the placebo group were lower than those previously reported in studies where there was a beneficial effect of a 5ARI, relative to placebo, on bleeding during TURP. PMID- 17407517 TI - Laparoscopic distal ureterectomy and anastomosis for management of low-risk upper urinary tract transitional cell carcinoma: preliminary results. AB - OBJECTIVE: To determine the surgical feasibility and early oncological outcomes of laparoscopic distal ureterectomy in patients with low-grade upper urinary tract transitional cell carcinoma (UUT-TCC). PATIENTS AND METHODS: We retrospectively reviewed patients treated laparoscopically with conservative management for a UUT-TCC between 2001 and 2005. We collected data on gender, age, mode of diagnosis, smoking, history of bladder cancer, complications, tumour site, size, stage, grade, hospital stay, recurrence and progression. RESULTS: Data were analysed for six patients with a mean (range) age of 68.5 (54-76) years. Four patients had a diagnostic ureteroscopy with biopsy. The operative duration was 173.3 (120-240) min, the estimated blood loss was 75 (50-200) mL and the length of ureteric resection was 5.23 cm. Two patients required a psoas hitch. JJ stents were maintained for 25.8 (15-30) days. The hospital stay was 6 (5-8) days. There were minor complications in three patients after surgery. The follow-up was 32 (17-46) months. The tumour size was 1.7 (0.8-2.6) cm. There were low-grade tumours in four patients and pTa in five. All patients are alive and free of disease; there were no anastomotic strictures. Two patients developed a recurrence, one in the ipsilateral renal pelvis and one in the bladder. CONCLUSION: Laparoscopic distal ureterectomy with direct re-implantation is technically feasible for low-risk UUT-TCC (i.e. low-grade, noninvasive), in the properly selected patient. Early oncological outcomes are promising but strict surveillance protocols must be followed. PMID- 17407518 TI - Laparoscopic inguinal hernia repair during laparoscopic radical prostatectomy. AB - OBJECTIVE: To describe our experience of simultaneous laparoscopic radical prostatectomy (LRP) and inguinal hernia repair (LIHR) with a non-absorbable mesh, as there are few reports of simultaneous herniorrhaphy during LRP. PATIENTS AND METHODS: Forty patients who had simultaneous LIHR and LRP were retrospectively reviewed. All operations were completed via antegrade techniques using a non absorbable mesh for the LIHR, as the results with absorbable mesh were disappointing. RESULTS: In all, 48 clinically apparent hernias were repaired in 40 patients (mean age 60 years). Of these, 13 were left-sided, 23 right-sided, and six bilateral; 19 were direct, 14 indirect, two pantaloon, three femoral, and in 10 the type was not recorded. The mean operative duration was 172 min and the mean hospital stay was 1.5 days. Two patients had a urine leak after surgery, which resolved with no further intervention, and two developed a pelvic lymphocele, one at 4 months and the other at 2 months after surgery. Two patients required urinary catheter re-insertion for retention after surgical catheter removal at 9 and 10 days after surgery, respectively. One patient developed a deep venous thrombosis 19 days after surgery. Of the 40 patients, 36 (90%) were followed for a mean of 10 months; none had a hernia recurrence on the repaired side, while two developed a new symptomatic contralateral hernia. CONCLUSIONS: LIHR is a successful and reliable way to treat symptomatic patients who are treated surgically for prostate cancer. PMID- 17407519 TI - Magnetic resonance imaging in the evaluation of pelvi-ureteric junction obstruction: an all-in-one approach. AB - OBJECTIVE: To study the ability of comprehensive magnetic resonance imaging (MRI) to replace multiple imaging methods in the evaluation of patients with pelvi ureteric junction obstruction (PUJO). PATIENTS AND METHODS: Between May 2003 and November 2005, 46 consecutive patients (22 male and 24 females; mean age 31.6 years) with symptomatic primary PUJO were included. All had comprehensive MRI, including MR urography (MRU), dynamic MRI and MR angiography (MRA). Morphological results of MRU were compared with that of renal ultrasonography or intravenous urography, while the anatomical results of MRA were compared with the operative findings. A correlation between MR clearance and radioisotope clearance was done using linear regression analysis. RESULTS: MRU showed the morphology of the collecting system in all patients, and the ureter below the PUJ in 31 of 46 (67%), but renal stones were missed in three of 10 patients. MRA showed crossing vessels in 22 patients (48%). There was a strong correlation between MR clearance and radioisotope clearance (r = 0.823, P < 0.001). From the MRI results, 35 patients (19 with crossing vessels and 16 with marked hydronephrosis) had pyeloplasty and 11 had an endopyelotomy. Findings during pyeloplasty showed one false-negative and one false-positive result of the preoperative MRI. Therefore, the sensitivity, specificity and accuracy of MRA were 95%, 94% and 94%, respectively. The outcome was successful in 44 (96%) patients. One failure after pyeloplasty was managed with endopyelotomy and the other was treated with pyeloplasty after endopyelotomy. CONCLUSION: Comprehensive MRI is a valuable and accurate single-imaging method for evaluating patients with PUJO. PMID- 17407520 TI - Influence of blood sampling on protein profiling and pattern analysis using matrix-assisted laser desorption/ionisation mass spectrometry. AB - OBJECTIVE: To describe the influence of blood sampling/sampling tubes on mass spectrometric and clustering results, and on clinical blood variables, in blood samples collected from healthy volunteers and patients with prostate cancer. PATIENTS, SUBJECTS AND METHODS: Two venous blood samples were taken from 12 healthy volunteers and 12 patients with localized prostate cancer. Two blood samples were taken from each participant using two different venepuncture systems (group A and group B). The Kolmogorov-Smirnov test was used to identify the peaks distinguishing the different groups. In a 10-fold cross-validation study, decision trees for identifying discriminatory peaks that separate the benign from the malignant were constructed. RESULTS: The decision tree separated samples measured by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) from healthy volunteers from those of patients with prostate cancer, with a sensitivity of 93.6% and a specificity of 91.6%. Of special interest is that one peak at 6941 m/z was produced during blood sample preparation and had a very powerful influence on the results of the classification. CONCLUSION: The results clearly showed that blood-sampling systems have a great influence on the recorded MALDI MS traces, and thus can markedly influence and confound the results of the MS analysis, whereas clinical variables might remain unchanged. MS profiling is a promising method of marker discovery, but as it could be shown well-designed studies are critical to allow proper interpretation for the identification of key variables as well as for the clinical use. PMID- 17407521 TI - The role of the urothelium in mediating bladder responses to isoprenaline. AB - OBJECTIVES: To investigate whether the responses of the pig bladder to isoprenaline (a nonselective beta-adrenoceptor agonist) are influenced by the presence of an intact urothelium and whether any influence might be attributed to the release of nitric oxide (NO), since stimulation of beta-adrenoceptors induces a direct relaxation of detrusor smooth muscle and beta-adrenoceptors are also present on the urothelium. MATERIAL AND METHODS: Paired (in the presence or absence of urothelium) longitudinal strips of pig bladder dome were set up in tissue baths and the developed tension recorded. Relaxation responses to isoprenaline were examined after pre-contraction with carbachol. The inhibitory effects of isoprenaline were examined by comparing responses to carbachol in the absence and presence of isoprenaline. To examine a possible role for NO, similar experiments were performed in the presence of the NO synthase inhibitor N(G) nitro-L-arginine (L-NNA). RESULTS: In the presence of the urothelium, both the potency (pEC(50)) and the maximum contractile responses to carbachol were depressed. In relaxation experiments, isoprenaline relaxed carbachol pre contracted tissues by approximately 75%, and the potency and maximum relaxation were similar in the absence and presence of the urothelium. In the inhibition experiments, the presence of isoprenaline caused rightward parallel shifts of the concentration-response curves to carbachol, but isoprenaline did not influence the maximum contractions. In the presence of the urothelium there was a greater shift with 0.1 microm isoprenaline than in denuded tissues. Incubation with L-NNA did not affect the influence of the urothelium on responses to isoprenaline in any experimental group. CONCLUSIONS: The relaxation responses of the bladder to isoprenaline do not appear to involve the urothelium or NO release in vitro. However, contractile responses to carbachol were inhibited in the presence of an intact urothelium, and this might reflect the release of an inhibitory factor other than NO. PMID- 17407522 TI - The effect of maturation and age on oestrogen-induced functional hypertrophy of the female rabbit bladder. AB - OBJECTIVE: To evaluate the effect of maturation and ageing on oestrogen-induced functional hypertrophy of the female rabbit bladder. MATERIALS AND METHODS: Twenty female rabbits were separated into two groups of 10 each by age, young (immature) and old rabbits and each age group was subdivided into three subgroups. The rabbits in subgroup 1 were controls, subgroup 2 were ovariectomized (Ovx) and subgroup 3 were Ovx and received 17-beta oestradiol (1 mg/kg/day) by a subcutaneous slow-release tablet implant. After 15 days of treatment, the rabbits were killed, the bladder was excised, and the body and base separated; two full-thickness longitudinal strips from the ventral surface of the bladder body, and one full-thickness strip from the base, were prepared for contractile studies. The contractile responses to electrical-field stimulation, carbachol, ATP and KCl were determined for both the bladder body and base strips. In addition, full-thickness strips of bladder body and base were fixed in formalin for histological and immunohistological studies. RESULTS: Ovx plus oestradiol resulted in significant increases in bladder weight and responses to all forms of stimulation in young and old rabbits (except for the response to KCl). Vascular density and the smooth muscle (SM)/collagen ratio significantly increased after oestradiol replacement. Interestingly, the increase in vascular density was greater in the young than in the old rabbits. CONCLUSIONS: The present study shows that oestrogen supplementation mediates a functional hypertrophy characterized by increased contractile responses to all forms of stimulation in both young and old rabbits. The increased contractile responses might be explained by the increases in vascular density and SM/collagen ratio. PMID- 17407523 TI - Slow progress in prostate cancer. PMID- 17407525 TI - Nurse-led flexible cystoscopy: experience from one UK centre. PMID- 17407526 TI - What are the risks of second cancer formation after radiotherapy to the prostate? PMID- 17407527 TI - Urethral stricture assessment: a prospective study evaluating urethral ultrasonography and conventional radiological studies. PMID- 17407528 TI - A prospective analysis of the diagnostic yield resulting from the attendance of 4020 patients at a protocol-driven haematuria clinic. PMID- 17407529 TI - Different brain effects during chronic and acute sacral neuromodulation in urge incontinent patients with implanted neurostimulators. PMID- 17407530 TI - Renal transplant-recipient surgery. PMID- 17407531 TI - Thellungiella: an Arabidopsis-related model plant adapted to cold temperatures. AB - Thellungiella salsuginea, a wild crucifer that grows in subarctic Canada and is closely related to Arabidopsis thaliana, was examined for its suitability as a model plant for studies of tolerance to cold and freezing temperatures. Thellungiella completed its life cycle at 5 degrees C, demonstrating that temperature-sensitive processes such as seed germination and the production of pollen and seeds were resistant to cold temperatures. Moreover, the plant exhibited dramatically different vegetative and flowering phenotypes in response to growth at cold temperature and shifts to cold temperature. Northern analyses showed that genes induced by cold in Arabidopsis, including CBF1, the transcriptional activator for the cold-regulated (COR) genes COR15a and COR47, were also expressed in Thellungiella. Freezing tolerance, assayed by the regrowth of intact plants, increased from -13.0 to -18.5 degrees C after cold treatment. The plants lacked endogenous ice nucleation or anti-freeze activity, indicating a potential for supercooling. As a close relative to Arabidopsis, Thellungiella exhibits extreme cold tolerance and should be an important model system in the elucidation of stress tolerance mechanisms. PMID- 17407532 TI - Evaporative enrichment and time lags between delta18O of leaf water and organic pools in a pine stand. AB - Understanding ecosystem water fluxes has gained increasing attention, as climate scenarios predict a drier environment for many parts of the world. Evaporative enrichment of (18)O (Delta(18)O) of leaf water and subsequent enrichment of plant organic matter can be used to characterize environmental and physiological factors that control evaporation, based on a recently established mechanistic model. In a Pinus sylvestris forest, we measured the dynamics of oxygen isotopic composition (delta(18)O) every 6 h for 4 d in atmospheric water vapour, xylem sap, leaf water and water-soluble organic matter in current (N) and previous year (N-1) needles, phloem sap, together with leaf gas exchange for pooled N and N-1 needles, and relevant micrometeorological variables. Leaf water delta(18)O showed strong diel periodicity, while delta(18)O in atmospheric water vapour and in xylem sap showed little variation. The Delta(18)O was consistently lower for N than for N-1 needles, possibly related to phenological stage. Modelled leaf water Delta(18)O showed good agreement with measured values when applying a non-steady state evaporative enrichment model including a Peclet effect. We determined the time lags between delta(18)O signals from leaf water to water-soluble foliar organic matter and to phloem sap at different locations down the trunk, which clearly demonstrated the relevance of considering these time-lag effects for carbon transport, source-sink and carbon flux partitioning studies. PMID- 17407533 TI - Guard-cell apoplastic sucrose concentration--a link between leaf photosynthesis and stomatal aperture size in the apoplastic phloem loader Vicia faba L. AB - In broad bean (Vicia faba L.), an apoplastic phloem loader, the sucrose concentration increases up to approximately 2 mM in the leaf apoplast and up to approximately 150 mM in the guard-cell apoplast during the photoperiod. This high concentration in the guard-cell apoplast results from transpiration and is sufficient osmotically to reduce stomatal aperture size by up to 3 microm or approximately 25% of the maximum aperture size. In this paper, we investigated a parallel and required role for high bulk-leaf apoplastic sucrose concentration, which correlates with high photosynthesis rate. An empirically determined combination of lowered light intensity and lowered CO(2) concentration reduced the photosynthesis rate to nominally one-fifth of the control value without a significant change in transpiration. This reduction in photosynthesis caused the sucrose concentration in the leaf apoplast--the immediate source pool for guard cells--to decrease by 70% (to 0.4 mM). In turn, sucrose concentration in the guard-cell apoplast decreased by approximately 80% (to approximately 40 mM). These results complete the required evidence for a non-exclusive, transpiration linked, photosynthesis-dependent passive mechanism for the modulation of stomatal aperture size. In an ancillary investigation, hexoses in the bulk-leaf apoplast decreased when photosynthesis was lowered, but their concentrations in the guard cell apoplast of control plants indicated that their osmotic contribution was negligible. PMID- 17407534 TI - Impacts of tree height on leaf hydraulic architecture and stomatal control in Douglas-fir. AB - This study investigated the mechanisms involved in the regulation of stomatal closure in Douglas-fir and evaluated the potential impact of compensatory adjustments in response to increasing tree height upon these mechanisms. In the laboratory, we measured leaf hydraulic conductance (K(leaf)) as leaf water potential (Psi(l)) declined for comparison with in situ diurnal patterns of stomatal conductance (g(s)) and Psi(l) in Douglas-fir across a height gradient, allowing us to infer linkages between diurnal changes in K(leaf) and g(s). A recently developed timed rehydration technique was used in conjunction with data from pressure-volume curves to develop hydraulic vulnerability curves for needles attached to small twigs. Laboratory-measured K(leaf) declined with increasing leaf water stress and was substantially reduced at Psi(l) values of -1.34, -1.45, -1.56 and -1.92 MPa for foliage sampled at mean heights of approximately 20, 35, 44 and 55 m, respectively. In situ g(s) measurements showed that stomatal closure was initiated at Psi(l) values of -1.21, -1.36, -1.74 and -1.86 MPa along the height gradient, which was highly correlated with Psi(l) values at loss of K(leaf). Cryogenic scanning electron microscopy (SEM) images showed that relative abundances of embolized tracheids in the central vein increased with increasing leaf water stress. Leaf embolism appeared to be coupled to changes in g(s) and might perform a vital function in stomatal regulation of plant water status and water transport in conifers. The observed trends in g(s) and K(leaf) in response to changes in Psi(l) along a height gradient suggest that the foliage at the tops of tall trees is capable of maintaining stomatal conductance at more negative Psi(l). This adaptation may allow taller trees to continue to photosynthesize during periods of greater water stress. PMID- 17407535 TI - Measurement of stem respiration of sycamore (Platanus occidentalis L.) trees involves internal and external fluxes of CO2 and possible transport of CO2 from roots. AB - CO(2) released by respiring cells in tree stems can either diffuse to the atmosphere or dissolve in xylem sap. In this study, the internal and external fluxes of CO(2) released from respiring stems of five sycamore (Platanus occidentalis L.) trees were calculated. Mean rates of stem respiration were highest in mid-afternoon and lowest at night, and were positively correlated with air temperature. Over a 24 h period, on average 34% of the CO(2) released by respiring cells in the measured stem segment remained within the tree. CO(2) efflux to the atmosphere consisted of similar proportions of CO(2) derived from local respiring cells (55%) and CO(2) that had been transported in the xylem (45%), indicating that CO(2) efflux does not accurately estimate respiration. A portion of the efflux of transported CO(2) appeared to have originated in the root system. A modification of the method for calculating stem respiration based on internal and external fluxes of CO(2) was developed to separate efflux due to local respiration from efflux of transported CO(2). PMID- 17407536 TI - Trade-off between root porosity and mechanical strength in species with different types of aerenchyma. AB - The objective of this work was to study the existence of a trade-off between aerenchyma formation and root mechanical strength. To this end, relationships among root anatomical traits and mechanical properties were analysed in plant species with contrasting root structural types: Paspalidium geminatum (graminaceous type), Cyperus eragrostis (cyperaceous type), Rumex crispus (Rumex type) and Plantago lanceolata (Apium type). Variations in anatomical traits and mechanical strength were assessed as a function of root diameter by exposing plants to 0, 7, 15 and 30 d of control and flooded conditions. For each species, the proportion of root cortex was positively associated with the increment of root diameter, contributing to the increase in root porosity under both control and flooded conditions. Moreover, cell lysis produced an additional increase in root porosity in most species under flooded conditions (except R. crispus). Both structural types that presented a uniseriate layer (epidermis) to cope with compression (Rumex and Apium types) were progressively weakened as root porosity increased. This effect was significant even when the increment of root porosity was solely because of increased root diameter (R. crispus), as when both processes (root diameter and cell lysis) added porosity to the roots (P. lanceolata). Conversely, structural types that presented a multiseriate ring of cells in the outer cortex (graminaceous and cyperaceous types) maintained mechanical strength over the whole range of porosity, in spite of lysogenic processes registered in the inner cortex. In conclusion, our study demonstrates a strong trade-off between aerenchyma formation and mechanical strength in root structural types that lacked a multiseriate ring of tissue for mechanical protection in the outer cortex. The results suggest that this ring of tissue plays a significant role in maintaining the mechanical strength of roots when flooding induces the generation of additional aerenchyma tissue in the root cortex. PMID- 17407537 TI - Regulation of brassinosteroid responses by phytochrome B in rice. AB - Plant growth and development are coordinately controlled by environmental signals and internal factors. Light signals, mediated by phytochromes, regulate photomorphogenesis by interacting with endogenous programmes that involve multiple phytohormones. Brassinosteroids (BRs) are a group of growth-promoting phytohormones with a crucial role in the light-dependent development of plants. However, the interaction between light-signalling pathways and BR signalling is not well understood. Here, we examined the responses of lamina joint inclination and coleoptile elongation to exogenous brassinolide (BL) under light or dark conditions. Both responses were more pronounced under darkness, implying that BR signalling is inhibited by light. To elucidate which phytochrome is involved in this interaction, we isolated rice phytochrome-deficient mutants (osphyA, osphyB and osphyC) from a T-DNA insertional population. Whereas the osphyA and osphyC knockout mutants did not differ from the wild-type plants in their BL responses, osphyB mutants were more sensitive. In addition, RT-PCR analysis revealed enhanced expression of BR-inducible genes and decreased transcript levels of BR biosynthetic genes in osphyB plants. These results suggest that Phytochrome B acts as a negative regulator of BL-regulated growth and development processes in rice. PMID- 17407538 TI - Variations in 13C discrimination during CO2 exchange by Picea sitchensis branches in the field. AB - We report diurnal variations in (13)C discrimination ((13)Delta) of Picea sitchensis (Bong.) Carr. branches measured in the field using a branch chamber technique. The observations were compared to predicted (13)Delta based on concurrent measurements of branch gas exchange. Observed (13)Delta values were described well by the classical model of (13)Delta including isotope effects during photorespiration, day respiration and CO(2) transfer through a series of resistances to the sites of carboxylation. A simplified linear of model (13)Delta did not capture the observed diurnal variability. At dawn and dusk, we measured very high (13)Delta values that were not predicted by either of the said models. Exploring the sensitivity of (13)Delta to possible respiratory isotope effects, we conclude that isotopic disequilibria between the gross fluxes of photosynthesis and day respiration can explain the high observed (13)Delta values during net photosynthetic gas exchange. Based on the classical model, a revised formulation incorporating an isotopically distinct substrate for day respiration was able to account well for the high observed dawn and dusk (13)Delta values. PMID- 17407539 TI - Characterization and expression analysis of genes encoding alpha and beta carbonic anhydrases in Arabidopsis. AB - Carbonic anhydrases (CAs) are Zn-containing metalloenzymes that catalyse the reversible hydration of CO(2). We investigated the alphaCA and betaCA families in Arabidopsis, which contain eight alphaCA (At alphaCA1-8) and six betaCA genes (At betaCA1-6). Analyses of expressed sequence tags (ESTs) from The Arabidopsis Information Resource (TAIR) database indicate that all the betaCA encoding sequences, but only three of the At alphaCA, are expressed. Using semi quantitative PCR experiments, functional CA genes were more strongly expressed in green tissue, but strong expression was also found in roots for betaCA3, betaCA6 and alphaCA2. Two alphaCA genes were shown to respond to the CO(2) environment, while the others were unresponsive. Using the green fluorescent reporter protein gene fused with cDNA sequences coding for betaCAs, we provided evidence that betaCAs were targeted to specific subcellular compartments: betaCA1 and betaCA5 were targeted to the chloroplast, betaCA2 and betaCA3 to the cytosol, betaCA4 to the plasma membrane and betaCA6 to the mitochondria. The targeting and the pattern of gene expression suggest that CA isoforms play specific roles in subcellular compartments, tissues and organs. The data indicate that other CA isoforms than the well-characterized betaCA1 may contribute to the CO(2) transfer in the cell to the catalytic site of ribulose 1.5-bisphosphate carboxylase/oxygenase (Rubisco). PMID- 17407540 TI - Transcriptional profiling of hexaploid wheat (Triticum aestivum L.) roots identifies novel, dehydration-responsive genes. AB - We used a long-oligonucleotide microarray to identify transcripts that increased or decreased in abundance in roots of dehydration-tolerant hexaploid bread wheat, in response to withholding of water. We observed that the major classes of dehydration-responsive genes (e.g. osmoprotectants, compatible solutes, proteases, glycosyltransferases/hydrolases, signal transducers components, ion transporters) were generally similar to those observed previously in other species and osmotic stresses. More specifically, we highlighted increases in transcript expression for specific genes including those putatively related to the synthesis of asparagine, trehalose, oligopeptide transporters, metal-binding proteins, the gamma-aminobutyric acid (GABA) shunt and transcription factors. Conversely, we noted a decrease in transcript abundance for diverse classes of glutathione and sulphur-related enzymes, specific amino acids, as well as MATE efflux carrier proteins. From these data, we identified a novel, dehydration induced putative AP2/ERF transcription factor, which we predict to function as a transcriptional repressor. We also identified a dehydration-induced 'little protein' (LitP; predicted mass: 8 kDa) that is highly conserved across spermatophytes. Using qRT-PCR, we compared the expression patterns of selected genes between two related wheat genotypes that differed in their susceptibility to dehydration, and confirmed that these novel genes were highly inducible by water limitation in both genotypes, although the magnitude of induction differed. PMID- 17407541 TI - Phototropic bending of non-elongating and radially growing woody stems results from asymmetrical xylem formation. AB - Active phototropic bending of non-elongating and radially growing portion of stems (woody stems) has not been previously documented, whereas negative gravitropic bending is well known. We found phototropic bending in woody stems and searched for the underlying mechanism. We inclined 1-year-old Quercus crispula Blume seedlings and unilaterally illuminated them from a horizontal direction perpendicular to ('normal' illumination) or parallel to ('parallel' illumination) the inclination azimuth. With normal illumination, active phototropic bending and xylem formation could be evaluated separately from the negative gravitropic response and vertical deflection resulting from the weight of the seedlings. One-year-old stems with normal illumination bent significantly, with asymmetrical xylem formation towards the illuminated upper surface and side of the stem, whereas those with parallel illumination showed non-significant lateral bending, with asymmetrical xylem formation only on the upper side. A mechanical model was built on the assumption that a bending moment resulted from the asymmetrical xylem formation during phototropic bending of the woody stems. The model fitted the relationship between the observed spatial distributions of the xylem and the observed lateral bending, and thus supported the hypothesis that phototropic bending of woody stems results from asymmetrical xylem formation, as such occurs during gravitropism. PMID- 17407542 TI - Isoprene synthase expression and protein levels are reduced under elevated O3 but not under elevated CO2 (FACE) in field-grown aspen trees. AB - Emission of hydrocarbons by trees has a crucial role in the oxidizing potential of the atmosphere. In particular, isoprene oxidation leads to the formation of tropospheric ozone and other secondary pollutants. It is expected that changes in the composition of the atmosphere will influence the emission rate of isoprene, which may in turn feedback on the accumulation of pollutants and greenhouse gases. We investigated the isoprene synthase (ISPS) gene expression and the ISPS protein levels in aspen trees exposed to elevated ozone (O(3)) and/or elevated carbon dioxide (CO(2)) in field-grown trees at the Aspen Free-Air Carbon Dioxide Enrichment (FACE) experimental site. Elevated O(3) reduced ISPS mRNA and the amount of ISPS protein in aspen leaves, whereas elevated CO(2) had no significant effect. Aspen clones with different O(3) sensitivity showed different levels of inhibition under elevated O(3) conditions. The drop in ISPS protein levels induced a drop in the isoprene emission rate under elevated O(3). However, the data indicated that other mechanisms also contributed to the observed strong inhibition of isoprene emission under elevated O(3). PMID- 17407543 TI - The relationship between isoprene emission rate and dark respiration rate in white poplar (Populus alba L.) leaves. AB - In past studies, it was hypothesized that reductions in chloroplast isoprene emissions at high atmospheric CO(2) concentrations were caused by competition between cytosolic and mitochondrial processes for the same substrate, possibly phosphoenolpyruvate (PEP). We conducted field and laboratory experiments using leaves of white poplar (Populus alba L.) to identify whether an inverse relationship occurs between the dark respiration rate (a mitochondrial process) and the isoprene emission rate. Field experiments that were carried out in a free air CO(2)-enriched (FACE) facility showed no clear effect of elevated CO(2) on either isoprene emission rate or respiration rate by leaves. In young, not yet fully expanded leaves, low isoprene emission and high dark respiration rates were measured in both ambient and elevated CO(2). In these leaves, isoprene emission was inversely correlated with dark respiration. It is possible to interpret from these results that, in young leaves, high rates of growth respiration compete with isoprene biosynthesis for the same substrate. However, it is also possible that the negative correlation reflects the contrasting reductions in growth respiration and increases in expression of the enzyme isoprene synthase at this final stage of leaf maturation. In contrast to our observations on young leaves, respiration rate and isoprene emission rate were positively correlated in older, fully expanded leaves (8 and 11 from apex). A positive correlation was also found between respiration rate and isoprene emission rate when these parameters were modulated using different ozone exposure, growth light intensity, growth temperature and exposure to different leaf temperatures in laboratory experiments. These data show that competition for substrate between isoprene biosynthesis and leaf respiration does not determine the rate of isoprene emission in most circumstances that affect both processes. A negative correlation was observed across all experiments between isoprene emission rate and the activity of phosphoenolpyruvate carboxylase (PEPc), a cytosolic enzyme that competes with isoprene biosynthesis for substrate. The cytosolic metabolite, PEP, occurs at a metabolic branch point from which substrate flows into three processes: (1) the production of pyruvate for mitochondrial respiration, (2) the production of oxaloacetate (OAA) by PEPc for anabolic support of mitochondrial respiration and (3) transport into the chloroplast to support chloroplastic demands for pyruvate, including isoprenoid biosynthesis. The results of our observations suggest that only the second process competes for substrate with isoprenoid synthesis, while the partitioning of PEP between mitochondrial respiration and chloroplast isoprenoid biosynthesis is controlled in a way that retains balance in substrate demand. PMID- 17407544 TI - Absolute quantitative real-time polymerase chain reaction for the measurement of human papillomavirus E7 mRNA in cervical cytobrush specimens. AB - BACKGROUND: Few reports of the utilization of an accurate, cost-effective means for measuring HPV oncogene transcripts have been published. Several papers have reported the use of relative quantitation or more expensive Taqman methods. Here, we report a method of absolute quantitative real-time PCR utilizing SYBR-green fluorescence for the measurement of HPV E7 expression in cervical cytobrush specimens. RESULTS: The construction of a standard curve based on the serial dilution of an E7-containing plasmid was the key for being able to accurately compare measurements between cervical samples. The assay was highly reproducible with an overall coefficient of variation of 10.4%. CONCLUSION: The use of highly reproducible and accurate SYBR-based real-time polymerase chain reaction (PCR) assays instead of performing Taqman-type assays allows low-cost, high-throughput analysis of viral mRNA expression. The development of such assays will help in refining the current screening programs for HPV-related carcinomas. PMID- 17407545 TI - Solvent exposure and malignant lymphoma: a population-based case-control study in Germany. AB - AIMS: To analyze the relationship between exposure to chlorinated and aromatic organic solvents and malignant lymphoma in a multi-centre, population-based case control study. METHODS: Male and female patients with malignant lymphoma (n = 710) between 18 and 80 years of age were prospectively recruited in six study regions in Germany (Ludwigshafen/Upper Palatinate, Heidelberg/Rhine-Neckar County, Wurzburg/Lower Frankonia, Hamburg, Bielefeld/Gutersloh, and Munich). For each newly recruited lymphoma case, a gender, region and age-matched (+/- 1 year of birth) population control was drawn from the population registers. In a structured personal interview, we elicited a complete occupational history, including every occupational period that lasted at least one year. On the basis of job task-specific supplementary questionnaires, a trained occupational physician assessed the exposure to chlorinated hydrocarbons (trichloroethylene, tetrachloroethylene, dichloromethane, carbon tetrachloride) and aromatic hydrocarbons (benzene, toluene, xylene, styrene). Odds ratios (OR) and 95% confidence intervals (CI) were calculated using conditional logistic regression analysis, adjusted for smoking (in pack years) and alcohol consumption. To increase the statistical power, patients with specific lymphoma subentities were additionally compared with the entire control group using unconditional logistic regression analysis. RESULTS: We observed a statistically significant association between high exposure to chlorinated hydrocarbons and malignant lymphoma (Odds ratio = 2.1; 95% confidence interval 1.1-4.3). In the analysis of lymphoma subentities, a pronounced risk elevation was found for follicular lymphoma and marginal zone lymphoma. When specific substances were considered, the association between trichloroethylene and malignant lymphoma was of borderline statistical significance. Aromatic hydrocarbons were not significantly associated with the lymphoma diagnosis. CONCLUSION: In accordance with the literature, this data point to a potential etiologic role of chlorinated hydrocarbons (particularly trichloroethylene) and malignant lymphoma. Chlorinated hydrocarbons might affect specific lymphoma subentities differentially. Our study does not support a strong association between aromatic hydrocarbons (benzene, toluene, xylene, or styrene) and the diagnosis of a malignant lymphoma. PMID- 17407546 TI - Evolutionary hierarchies of conserved blocks in 5'-noncoding sequences of dicot rbcS genes. AB - BACKGROUND: Evolutionary processes in gene regulatory regions are major determinants of organismal evolution, but exceptionally challenging to study. We explored the possibilities of evolutionary analysis of phylogenetic footprints in 5'-noncoding sequences (NCS) from 27 ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) genes, from three dicot families (Brassicaceae, Fabaceae and Solanaceae). RESULTS: Sequences of up to 400 bp encompassing proximal promoter and 5'-untranslated regions were analyzed. We conducted phylogenetic footprinting by several alternative methods: generalized Lempel-Ziv complexity (CLZ), multiple alignments with DIALIGN and ALIGN-M, and the MOTIF SAMPLER Gibbs sampling algorithm. These tools collectively defined 36 conserved blocks of mean length 12.8 bp. On average, 12.5 blocks were found in each 5'-NCS. The blocks occurred in arrays whose relative order was absolutely conserved, confirming the existence of 'conserved modular arrays' in promoters. Identities of half of the blocks confirmed past rbcS research, including versions of the I-box, G-box, and GT-1 sites such as Box II. Over 90% of blocks overlapped DNase-protected regions in tomato 5'-NCS. Regions characterized by low CLZ in sliding-window analyses were also frequently associated with DNase-protection. Blocks could be assigned to evolutionary hierarchies based on taxonomic distribution and estimated age. Lineage divergence dates implied that 13 blocks found in all three plant families were of Cretaceous antiquity, while other family-specific blocks were much younger. Blocks were also dated by formation of multigene families, using genome and coding sequence information. Dendrograms of evolutionary relations of the 5' NCS were produced by several methods, including: cluster analysis using pairwise CLZ values; evolutionary trees of DIALIGN sequence alignments; and cladistic analysis of conserved blocks. CONCLUSION: Dicot 5'-NCS contain conserved modular arrays of recurrent sequence blocks, which are coincident with functional elements. These blocks are amenable to evolutionary interpretation as hierarchies in which ancient, taxonomically widespread blocks can be distinguished from more recent, taxon-specific ones. PMID- 17407547 TI - Porcine transcriptome analysis based on 97 non-normalized cDNA libraries and assembly of 1,021,891 expressed sequence tags. AB - BACKGROUND: Knowledge of the structure of gene expression is essential for mammalian transcriptomics research. We analyzed a collection of more than one million porcine expressed sequence tags (ESTs), of which two-thirds were generated in the Sino-Danish Pig Genome Project and one-third are from public databases. The Sino-Danish ESTs were generated from one normalized and 97 non normalized cDNA libraries representing 35 different tissues and three developmental stages. RESULTS: Using the Distiller package, the ESTs were assembled to roughly 48,000 contigs and 73,000 singletons, of which approximately 25% have a high confidence match to UniProt. Approximately 6,000 new porcine gene clusters were identified. Expression analysis based on the non-normalized libraries resulted in the following findings. The distribution of cluster sizes is scaling invariant. Brain and testes are among the tissues with the greatest number of different expressed genes, whereas tissues with more specialized function, such as developing liver, have fewer expressed genes. There are at least 65 high confidence housekeeping gene candidates and 876 cDNA library specific gene candidates. We identified differential expression of genes between different tissues, in particular brain/spinal cord, and found patterns of correlation between genes that share expression in pairs of libraries. Finally, there was remarkable agreement in expression between specialized tissues according to Gene Ontology categories. CONCLUSION: This EST collection, the largest to date in pig, represents an essential resource for annotation, comparative genomics, assembly of the pig genome sequence, and further porcine transcription studies. PMID- 17407548 TI - The regulation of cyclin D1 degradation: roles in cancer development and the potential for therapeutic invention. AB - Cyclin D1 is an important regulator of cell cycle progression and can function as a transcriptionl co-regulator. The overexpression of cyclin D1 has been linked to the development and progression of cancer. Deregulated cyclin D1 degradation appears to be responsible for the increased levels of cyclin D1 in several cancers. Recent findings have identified novel mechanisms involved in the regulation of cyclin D1 stability. A number of therapeutic agents have been shown to induce cyclin D1 degradation. The therapeutic ablation of cyclin D1 may be useful for the prevention and treatment of cancer. In this review, current knowledge on the regulation of cyclin D1 degradation is discussed. Novel insights into cyclin D1 degradation are also discussed in the context of ablative therapy. A number of unresolved questions regarding the regulation of cellular cyclin D1 levels are also addressed. PMID- 17407549 TI - Chromatin remodeling agent trichostatin A: a key-factor in the hepatic differentiation of human mesenchymal stem cells derived of adult bone marrow. AB - BACKGROUND: The capability of human mesenchymal stem cells (hMSC) derived of adult bone marrow to undergo in vitro hepatic differentiation was investigated. RESULTS: Exposure of hMSC to a cocktail of hepatogenic factors [(fibroblast growth factor-4 (FGF-4), hepatocyte growth factor (HGF), insulin-transferrin sodium-selenite (ITS) and dexamethasone)] failed to induce hepatic differentiation. Sequential exposure to these factors (FGF-4, followed by HGF, followed by HGF+ITS+dexamethasone), however, resembling the order of secretion during liver embryogenesis, induced both glycogen-storage and cytokeratin (CK)18 expression. Additional exposure of the cells to trichostatin A (TSA) considerably improved endodermal differentiation, as evidenced by acquisition of an epithelial morphology, chronological expression of hepatic proteins, including hepatocyte nuclear factor (HNF)-3beta, alpha-fetoprotein (AFP), CK18, albumin (ALB), HNF1alpha, multidrug resistance-associated protein (MRP)2 and CCAAT-enhancer binding protein (C/EBP)alpha, and functional maturation, i.e. upregulated ALB secretion, urea production and inducible cytochrome P450 (CYP)-dependent activity. CONCLUSION: hMSC are able to undergo mesenchymal-to-epithelial transition. TSA is hereby essential to promote differentiation of hMSC towards functional hepatocyte-like cells. PMID- 17407550 TI - Sickness absence poses a threat to the Swedish Welfare State: a cross-sectional study of sickness absence and self-reported illness. AB - BACKGROUND: The increasing cost of public social sickness insurance poses a serious economic threat to the Swedish welfare state. In recent years, expenditures for social insurance in general, as well as social sickness insurance in particular, have risen steeply in Sweden. This cross-sectional study analyzed the association between sickness absence (SA) and self-reported reduced working capacity due to a longstanding illness (>3 months), as well between SA and a number of other health problems. METHODS: Self-reported data on longstanding illness and resultant reduced working capacity, socioeconomic factors, working environment, psychosomatic complaints, anxiety, and general health were obtained for 22,281 employed (paid) persons aged 25 to 64 years. These data were retrieved from the Swedish Living Conditions Survey for 1995 to 2002. National civic registration numbers, replaced with serial numbers to ensure anonymity, were used to link these data to individual-level SA records from the National Social Insurance Board. A logistic regression model was used to estimate the odds ratio of the main outcome variable for the three levels of the SA variable (0-28, 29-90, >90 days/year). RESULTS: There was an obvious increasing gradient in length of SA and increasing odds of reporting reduced working capacity. Odds ratios ranged from 3.5 to 19.0; i.e., those with more than ninety days of SA had 19.0 times higher odds of reporting reduced working capacity than those with 0-28 days of SA a year. This very strong association changed less than 10% after adjusting for demographic, socioeconomic, and working environment characteristics. A total of 48.7% of persons on sick leave > or = 29 days reported no longstanding illness and reduced working capacity. Of these persons, about 43% reported one or more other health problem. CONCLUSION: We confirmed that longstanding illness that results in self-reported reduced working capacity is an important variable related to length of SA, even after taking important confounders into consideration. We found a little less than half of those on sick leave reported no reduced working capacity due to longstanding illness, and some of these reported no other health problems. However, it is possible that some respondents had health problems not captured in the survey. PMID- 17407551 TI - Intra- and extra-familial adverse childhood experiences and a history of childhood psychosomatic disorders among Japanese university students. AB - BACKGROUND: Japan has been witnessing a considerable increase in the number of children with psychosomatic disorders. The purpose of this study is to examine the relationship between the risk of psychosomatic disorder in adolescents and intra- and extra-familial adverse childhood experiences (ACEs). METHODS: A retrospective cohort study of 1592 Japanese university students (52% male, mean age 19.9 years) who completed a survey about intra- and extra-familial ACEs and the incidence of childhood psychosomatic disorders. Intra-familial ACEs included domestic violence, physical violence, emotional abuse, illness in household, parental divorce, no parental affection, and dysfunctional family. Extra-familial ACEs included physical violence or negative recognition by teachers, being bullied in elementary or junior high school, or sexual violence. RESULTS: The frequency of psychosomatic disorders among the respondents was 14.8%. Among the 7 intra-familial ACEs, emotional abuse (relative risk, RR = 1.9) and illness in household (RR = 1.7) increased the risk of psychosomatic disorders. Estimates of the relative risk for the 5 extra-familial ACEs were statistically significant and ranged from 1.5 for being bullied in elementary school or physical violence from teachers to 2.4. Students who had 3 or more intra-familial ACEs and 2 or more extra-familial ACEs had a 3.0 relative risk for psychosomatic disorder. CONCLUSION: These results suggest that intra- and extra-familial ACEs are associated with the development of psychosomatic disorders. Therefore, sufficient evaluation of ACEs should be performed in adolescent patients with psychosomatic disorder. PMID- 17407552 TI - A TATA binding protein regulatory network that governs transcription complex assembly. AB - BACKGROUND: Eukaryotic genes are controlled by proteins that assemble stepwise into a transcription complex. How the individual biochemically defined assembly steps are coordinated and applied throughout a genome is largely unknown. Here, we model and experimentally test a portion of the assembly process involving the regulation of the TATA binding protein (TBP) throughout the yeast genome. RESULTS: Biochemical knowledge was used to formulate a series of coupled TBP regulatory reactions involving TFIID, SAGA, NC2, Mot1, and promoter DNA. The reactions were then linked to basic segments of the transcription cycle and modeled computationally. A single framework was employed, allowing the contribution of specific steps to vary from gene to gene. Promoter binding and transcriptional output were measured genome-wide using ChIP-chip and expression microarray assays. Mutagenesis was used to test the framework by shutting down specific parts of the network. CONCLUSION: The model accounts for the regulation of TBP at most transcriptionally active promoters and provides a conceptual tool for interpreting genome-wide data sets. The findings further demonstrate the interconnections of TBP regulation on a genome-wide scale. PMID- 17407553 TI - Infection of hepatitis B virus in extrahepatic endothelial tissues mediated by endothelial progenitor cells. AB - BACKGROUND: Hepatitis B virus (HBV) replication has been reported to be involved in many extrahepatic viral disorders; however, the mechanism by which HBV is trans-infected into extrahepatic tissues such as HBV associated myocarditis remains largely unknown. RESULTS: In this study, we showed that human cord blood endothelial progenitor cells (EPCs), but not human umbilical vein endothelial cells (HUVECs) could be effectively infected by uptake of HBV in vitro. Exposure of EPCs with HBV resulted in HBV DNA and viral particles were detected in EPCs at day 3 after HBV challenge, which were peaked around day 7 and declined in 3 weeks. Consistently, HBV envelope surface and core antigens were first detected in EPCs at day 3 after virus challenge and were retained to be detectable for 3 weeks. In contrast, HBV covalently closed circular DNA was not detected in EPCs at any time after virus challenge. Intravenous transplantation of HBV-treated EPCs into myocardial infarction and acute renal ischemia mouse model resulted in incorporation of HBV into injured heart, lung, and renal capillary endothelial tissues. CONCLUSION: These results strongly support that EPCs serve as virus carrier mediating HBV trans-infection into the injured endothelial tissues. The findings might provide a novel mechanism for HBV-associated myocarditis and other HBV-related extrahepatic diseases as well. PMID- 17407554 TI - A new method to transfect the hypoblast of the chick embryo reveals conservation of the regulation of an Otx2 enhancer between mouse and chick extraembryonic endoderm. AB - BACKGROUND: The mouse anterior visceral endoderm (AVE) and the chick hypoblast are thought to have homologous roles in the early stages of neural induction and primitive streak formation. In mouse, many regulatory elements directing gene expression to the AVE have been identified. However, there is no technique to introduce DNA into the chick hypoblast that would enable a comparison of their activity and this has hampered a direct comparison of the regulation of gene expression in the mouse and chick extraembryonic endoderm. RESULTS: Here we describe a new method to introduce DNA into the chick hypoblast, using lipofectamine-mediated transfection. We show that the hypoblast can be easily transfected and that it starts to express a luciferase reporter within 2 hours of transfection. The validity of technique is tested by following the movement and fate of hypoblast cells, which reveals their translocation to the anterior germinal crescent. We then introduce a vector containing GFP driven by the mouse VEcis-Otx2 enhancer (which directs gene expression to the mouse AVE) and we detect activity in the hypoblast. CONCLUSION: The new technique for delivering expression constructs to the chick hypoblast will enable studies on gene activity and regulation to be performed in this tissue, which has proved difficult to transfect by electroporation. Our findings also reveal that regulatory elements that direct gene expression to the mouse AVE are active in chick hypoblast, supporting the idea that these two tissues have homologous functions. PMID- 17407555 TI - Diagnosis and treatment of malaria in peripheral health facilities in Uganda: findings from an area of low transmission in south-western Uganda. AB - BACKGROUND: Early recognition of symptoms and signs perceived as malaria are important for effective case management, as few laboratories are available at peripheral health facilities. The validity and reliability of clinical signs and symptoms used by health workers to diagnose malaria were assessed in an area of low transmission in south-western Uganda. METHODS: The study had two components: 1) passive case detection where all patients attending the out patient clinic with a febrile illness were included and 2) a longitudinal active malaria case detection survey was conducted in selected villages. A malaria case was defined as any slide-confirmed parasitaemia in a person with an axillary temperature > or = 37.5 degrees C or a history of fever within the last 24 hrs and no signs suggestive of other diseases. RESULTS: Cases of malaria were significantly more likely to report joint pains, headache, vomiting and abdominal pains. However, due to the low prevalence of malaria, the predictive values of these individual signs alone, or in combination, were poor. Only 24.8% of 1627 patients had malaria according to case definition and > 75% of patients were unnecessarily treated for malaria and few slide negative cases received alternative treatment. CONCLUSION: In low-transmission areas, more attention needs to be paid to differential diagnosis of febrile illnesses In view of suggested changes in anti malarial drug policy, introducing costly artemisinin combination therapy accurate, rapid diagnostic tools are necessary to target treatment to people in need. PMID- 17407556 TI - IL-1beta induces murine airway 5-HT2A receptor hyperresponsiveness via a non transcriptional MAPK-dependent mechanism. AB - BACKGROUND: Interleukin 1 beta (IL-1beta) is found in bronchoalveolar lavage fluids from asthmatic patients and plays an important role in normal immunoregulatory processes but also in pathophysiological inflammatory responses. The present study was designed to investigate if IL-1beta could be involved in the development of airway hyperresponsiveness and if transcriptional mechanisms, epithelium contractile factors and mitogen-activated protein kinase (MAPK) pathways are involved in IL-1beta effect. METHODS: The effect of IL-1beta on 5 hydroxytryptamine (5-HT) induced bronchoconstriction was evaluated in an in-vitro model for assessment of long-term effects of inflammatory mediators on the airway smooth muscle. Murine tracheal segments were cultured up to 8 days in the absence or presence of IL-1beta with subsequent evaluation in a myograph system, along with mRNA quantification, focusing on the role of the epithelium, acetylcholine release, transcriptional mechanisms and MAPK activity. RESULTS: During control conditions, 5-HT induced a relatively weak contraction. Presence of IL-1beta increased this response in a time- and concentration-dependent way. The increased concentration-effect curves could be shifted rightwards in a parallel manner by ketanserin, a selective 5-HT2A receptor antagonist, indicating that the responses are mediated by 5-HT2A receptors. The mRNA levels of 5-HT2A receptors were not changed as a consequence of the IL-1beta treatment and actinomycin D, a general transcriptional inhibitor, failed to affect the contractile response, suggesting a non-transcriptional mechanism behind this phenomenon. Neither the removal of the epithelium nor the addition of atropine affected the IL-1beta induced enhancement of 5-HT2A receptor-mediated contractile response. Application of inhibitors for c-Jun N-terminal kinase (JNK), p38 and extracellular signal regulated kinase 1 and 2 (ERK1/2) showed that the signaling pathways for JNK and ERK1/2 dominated only in cultured segments (control) whereas JNK and p38 dominated in segments treated with IL-1beta. CONCLUSION: IL-1beta induces murine airway hyperresponsiveness, via a non-transcriptional up-regulation of 5-HT2A receptor-mediated contractile response. The increase of 5-HT contraction is unrelated to epithelial and cholinergic factors, but is dependent on IL-1beta induced changes of MAPK pathways. The fact that IL-1beta can alter airway responses to contractile agents such as 5-HT, via alteration of the intracellular MAPK signal transduction pathways, might provide a new concept for future treatment of asthma. PMID- 17407557 TI - The effects of alcohol consumption, psychological distress and smoking status on emergency department presentations in New South Wales, Australia. AB - BACKGROUND: Despite clear links between risky alcohol consumption, mental health problems and smoking with increased morbidity and mortality, there is inconclusive evidence about how these risk factors combine and if they are associated with increased attendance at emergency departments. This paper examines the population-level associations and interactions between alcohol consumption, psychological distress and smoking status with having presented to an emergency department in the last 12 months. METHODS: This study uses data from a representative sample of 34,974 participants aged 16 years and over from the New South Wales Population Health Survey, administered between 2002 and 2004. Statistical analysis included univariate statistics, cross-tabulations, and the estimation of prevalence rate ratios using Cox's proportional hazard regression model. RESULTS: Results show that high-risk alcohol consumption, high psychological distress and current smoking were all significantly and independently associated with a greater likelihood of presenting to an emergency department in the last year. Presenting to an emergency department was found to be three times more likely for women aged 30 to 59 years with all three risk factors and ten times more likely for women aged 60 years or more who reported high risk alcohol consumption and high psychological distress than women of these age groups without these risk factors. For persons aged 16 to 29 years, having high-risk alcohol consumption and being a current smoker doubles the risk of presenting to an emergency department. CONCLUSION: The combination of being a high-risk consumer of alcohol, having high psychological distress, and being a current smoker are associated with increased presentations to emergency departments, independent of age and sex. Further research is needed to enhance recognition of and intervention for these symptoms in an emergency department setting in order to improve patient health and reduce future re-presentations to emergency departments. PMID- 17407558 TI - Contribution of oncoproteomics to cancer biomarker discovery. AB - Oncoproteomics is the study of proteins and their interactions in a cancer cell by proteomic technologies. Proteomic research first came to the fore with the introduction of two-dimensional gel electrophoresis. At the turn of the century, proteomics has been increasingly applied to cancer research with the wide-spread introduction of mass spectrometry and proteinchip. There is an intense interest in applying proteomics to foster an improved understanding of cancer pathogenesis, develop new tumor biomarkers for diagnosis, and early detection using proteomic portrait of samples. Oncoproteomics has the potential to revolutionize clinical practice, including cancer diagnosis and screening based on proteomic platforms as a complement to histopathology, individualized selection of therapeutic combinations that target the entire cancer-specific protein network, real-time assessment of therapeutic efficacy and toxicity, and rational modulation of therapy based on changes in the cancer protein network associated with prognosis and drug resistance. Besides, oncoproteomics is also applied to the discovery of new therapeutic targets and to the study of drug effects. In pace with the successful completion of the Human Genome Project, the wave of proteomics has raised the curtain on the postgenome era. The study of oncoproteomics provides mankind with a better understanding of neoplasia. In this article, the discovery of cancer biomarkers in recent years is reviewed. The challenges ahead and perspectives of oncoproteomics for biomarkers development are also addressed. With a wealth of information that can be applied to a broad spectrum of biomarker research projects, this review serves as a reference for biomarker researchers, scientists working in proteomics and bioinformatics, oncologists, pharmaceutical scientists, biochemists, biologists, and chemists. PMID- 17407559 TI - Potential cost savings with terrestrial rabies control. AB - BACKGROUND: The cost-benefit of raccoon rabies control strategies such as oral rabies vaccination (ORV) are under evaluation. As an initial quantification of the potential cost savings for a control program, the collection of selected rabies cost data was pilot tested for five counties in New York State (NYS) in a three-year period. METHODS: Rabies costs reported to NYS from the study counties were computerized and linked to a human rabies exposure database. Consolidated costs by county and year were averaged and compared. RESULTS: Reported rabies associated costs for all rabies variants totalled $2.1 million, for human rabies postexposure prophylaxes (PEP) (90.9%), animal specimen preparation/shipment to laboratory (4.7%), and pet vaccination clinics (4.4%). The proportion that may be attributed to raccoon rabies control was 37% ($784,529). Average costs associated with the raccoon variant varied across counties from $440 to $1,885 per PEP, $14 to $44 per specimen, and $0.33 to $15 per pet vaccinated. CONCLUSION: Rabies costs vary widely by county in New York State, and were associated with human population size and methods used by counties to estimate costs. Rabies cost variability must be considered in developing estimates of possible ORV-related cost savings. Costs of PEPs and specimen preparation/shipments, as well as the costs of pet vaccination provided by this study may be valuable for development of more realistic scenarios in economic modelling of ORV costs versus benefits. PMID- 17407560 TI - Using a summary measure for multiple quality indicators in primary care: the Summary QUality InDex (SQUID). AB - BACKGROUND: Assessing the quality of primary care is becoming a priority in national healthcare agendas. Audit and feedback on healthcare quality performance indicators can help improve the quality of care provided. In some instances, fewer numbers of more comprehensive indicators may be preferable. This paper describes the use of the Summary Quality Index (SQUID) in tracking quality of care among patients and primary care practices that use an electronic medical record (EMR). All practices are part of the Practice Partner Research Network, representing over 100 ambulatory care practices throughout the United States. METHODS: The SQUID is comprised of 36 process and outcome measures, all of which are obtained from the EMR. This paper describes algorithms for the SQUID calculations, various statistical properties, and use of the SQUID within the context of a multi-practice quality improvement (QI) project. RESULTS: At any given time point, the patient-level SQUID reflects the proportion of recommended care received, while the practice-level SQUID reflects the average proportion of recommended care received by that practice's patients. Using quarterly reports, practice- and patient-level SQUIDs are provided routinely to practices within the network. The SQUID is responsive, exhibiting highly significant (p < 0.0001) increases during a major QI initiative, and its internal consistency is excellent (Cronbach's alpha = 0.93). Feedback from physicians has been extremely positive, providing a high degree of face validity. CONCLUSION: The SQUID algorithm is feasible and straightforward, and provides a useful QI tool. Its statistical properties and clear interpretation make it appealing to providers, health plans, and researchers. PMID- 17407561 TI - High abundance synovial fluid proteome: distinct profiles in health and osteoarthritis. AB - The development of increasingly high-throughput and sensitive mass spectroscopy based proteomic techniques provides new opportunities to examine the physiology and pathophysiology of many biologic fluids and tissues. The purpose of this study was to determine protein expression profiles of high-abundance synovial fluid (SF) proteins in health and in the prevalent joint disease osteoarthritis (OA). A cross-sectional study of 62 patients with early OA (n = 21), patients with late OA (n = 21), and control individuals (n = 20) was conducted. SF proteins were separated by using one-dimensional PAGE, and the in-gel digested proteins were analyzed by electrospray ionization tandem mass spectrometry. A total of 362 spots were examined and 135 high-abundance SF proteins were identified as being expressed across all three study cohorts. A total of 135 SF proteins were identified. Eighteen proteins were found to be significantly differentially expressed between control individuals and OA patients. Two subsets of OA that are not dependent on disease duration were identified using unsupervised analysis of the data. Several novel SF proteins were also identified. Our analyses demonstrate no disease duration-dependent differences in abundant protein composition of SF in OA, and we clearly identified two previously unappreciated yet distinct subsets of protein profiles in this disease cohort. Additionally, our findings reveal novel abundant protein species in healthy SF whose functional contribution to SF physiology was not previously recognized. Finally, our studies identify candidate biomarkers for OA with potential for use as highly sensitive and specific tests for diagnostic purposes or for evaluating therapeutic response. PMID- 17407562 TI - Rosai-Dorfman disease: a previously unreported association with sickle cell disease. AB - BACKGROUND: Rosai-Dorfman Disease is an uncommon benign systemic histio proliferative disease. This is the first time the disease, although more common in people of African descent, is described in association with Sickle cell disease. CASE PRESENTATION: A Nigerian boy born started a complex medical history with post-natal anemia of unknown origin. Subsequently he was diagnosed with Sickle Cell Anemia (Hb SS). At age 3 during a routine review, he was noted to have generalised massive lymphadenopathy. He had further reoccurrences of this lymphadenopathy, but investigations did not reveal the cause until age five. At this point, because of the progressive lymph node enlargement, a biopsy was performed, and he was diagnosed with Rosai-Dorfman Disease. Since that time, the child has had further episodes of intermittent massive lymphadenopathy, particularly associated with Sickle Crisis. His medical history has been further complicated by development of complications from Sickle Cell Disease, cardiomyopathy and an autoimmune hemolytic anemia with multiple alloantibodies. CONCLUSION: This case for the first time presents the co-existence of two diseases, of increased prevalence in those of African descent, but to date not described in the literature to occur concurrently. PMID- 17407563 TI - Influenza pandemic: perception of risk and individual precautions in a general population. Cross sectional study. AB - BACKGROUND: An influenza pandemic may have considerable impact on health and societal functioning. The aim of this study was to explore people's reflections on the consequences of a pandemic. METHODS: Cross-sectional web-based survey of 1,168 Norwegians aged 16-82 years. The main outcome measures were answers to questions about a potential pandemic ("serious influenza epidemic"): statements about personal precautions including stockpiling Tamiflu, the perceived number of fatalities, the perceived effects of Tamiflu, the sources of information about influenza and trust in public information. RESULTS: While 80% of the respondents stated that they would be "careful about personal hygiene", only a few would stay away from work (2%), or move to an isolated place (4%). While 27% of respondents were uncertain about the number of fatalities during an influenza pandemic, 48% thought it would be lower than the estimate of Norwegian health authorities (0.05%-1%) and only 3% higher. At least half of the respondents thought that Tamiflu might reduce the mortality risk, but less than 1% had personally purchased the drug. The great majority had received their information from the mass media, and only 9% directly from health authorities. Still the majority (65%) trusted information from the authorities, and only 9% reported overt distrust. CONCLUSION: In Norway, considerable proportions of people seem to consider the mortality risk during a pandemic less than health authorities do. Most people seem to be prepared to take some, but not especially disruptive, precautions. PMID- 17407564 TI - Relatedness between the two-component lantibiotics lacticin 3147 and staphylococcin C55 based on structure, genetics and biological activity. AB - BACKGROUND: Two component lantibiotics, such as the plasmid-encoded lacticin 3147 produced by Lactococcus lactis DPC3147 and staphylococcin C55 produced by Staphylococcus aureus C55, represent an emerging subgroup of bacteriocins. These two bacteriocins are particularly closely related, exhibiting 86% (LtnA1 and C55alpha) and 55% (LtnA2 and C55beta) identity in their component peptides. The aim of this study was to investigate, for the first time for any two component bacteriocins, the significance of the relatedness between these two systems. RESULTS: So close is this relatedness that the hybrid peptide pairs LtnA1:C55beta and C55alpha:LtnA2 were found to have activities in the single nanomolar range, comparing well with the native pairings. To determine whether this flexibility extended to the associated post-translational modification/processing machinery, the staphylococcin C55 structural genes were directly substituted for their lacticin 3147 counterparts in the ltn operon on the large conjugative lactococcal plasmid pMRC01. It was established that the lacticin LtnA1 post-translational and processing machinery could produce functionally active C55alpha, but not C55beta. In order to investigate in closer detail the significance of the differences between LtnA1 and C55alpha, three residues in LtnA1 were replaced with the equivalent residues in C55alpha. Surprisingly, one such mutant LtnA1-Leu21Ala was not produced. This may be significant given the positioning of this residue in a putative lipid II binding loop. CONCLUSION: It is apparent, despite sharing striking similarities in terms of structure and activity, that these two complex bacteriocins display some highly dedicated features particular to either system. PMID- 17407565 TI - Conformational study of the protegrin-1 (PG-1) dimer interaction with lipid bilayers and its effect. AB - BACKGROUND: Protegrin-1 (PG-1) is known as a potent antibiotic peptide; it prevents infection via an attack on the membrane surface of invading microorganisms. In the membrane, the peptide forms a pore/channel through oligomerization of multiple subunits. Recent experimental and computational studies have increasingly unraveled the molecular-level mechanisms underlying the interactions of the PG-1 beta-sheet motifs with the membrane. The PG-1 dimer is important for the formation of oligomers, ordered aggregates, and for membrane damaging effects. Yet, experimentally, different dimeric behavior has been observed depending on the environment: antiparallel in the micelle environment, and parallel in the POPC bilayer. The experimental structure of the PG-1 dimer is currently unavailable. RESULTS: Although the beta-sheet structures of the PG-1 dimer are less stable in the bulk water environment, the dimer interface is retained by two intermolecular hydrogen bonds. The formation of the dimer in the water environment implies that the pathway of the dimer invasion into the membrane can originate from the bulk region. In the initial contact with the membrane, both the antiparallel and parallel beta-sheet conformations of the PG-1 dimer are well preserved at the amphipathic interface of the lipid bilayer. These beta-sheet structures illustrate the conformations of PG-1 dimer in the early stage of the membrane attack. Here we observed that the activity of PG-1 beta sheets on the bilayer surface is strongly correlated with the dimer conformation. Our long-term goal is to provide a detailed mechanism of the membrane-disrupting effects by PG-1 beta-sheets which are able to attack the membrane and eventually assemble into the ordered aggregates. CONCLUSION: In order to understand the dimeric effects leading to membrane damage, extensive molecular dynamics (MD) simulations were performed for the beta-sheets of the PG-1 dimer in explicit water, salt, and lipid bilayers composed of POPC lipids. Here, we studied PG-1 dimers when organized into a beta-sheet motif with antiparallel and parallel beta sheet arrangements in an NCCN packing mode. We focus on the conformations of PG-1 dimers in the lipid bilayer, and on the correlation between the conformations and the membrane disruption effects by PG-1 dimers. We investigate equilibrium structures of the PG-1 dimers in different environments in the early stage of the dimer invasion. The dimer interface of the antiparallel beta-sheets is more stable than the parallel beta-sheets, similar to the experimental observation in micelle environments. However, we only observe membrane disruption effects by the parallel beta-sheets of the PG-1 dimer. This indicates that the parallel beta sheets interact with the lipids with the beta-sheet plane lying obliquely to the bilayer surface, increasing the surface pressure in the initial insertion into the lipid bilayer. Recent experimental observation verified that parallel PG-1 dimer is biologically more active to insert into the POPC lipid bilayer. PMID- 17407566 TI - Maximum occlusal force and medial mandibular flexure in relation to vertical facial pattern: a cross-sectional study. AB - BACKGROUND: Vertical facial pattern may be related to the direction of pull of the masticatory muscles, yet its effect on occlusal force and elastic deformation of the mandible still is unclear. This study tested whether the variation in vertical facial pattern is related to the variation in maximum occlusal force (MOF) and medial mandibular flexure (MMF) in 51 fully-dentate adults. METHODS: Data from cephalometric analysis according to the method of Ricketts were used to divide the subjects into three groups: Dolichofacial (n = 6), Mesofacial (n = 10) and Brachyfacial (n = 35). Bilateral MOF was measured using a cross-arch force transducer placed in the first molar region. For MMF, impressions of the mandibular occlusal surface were made in rest (R) and in maximum opening (O) positions. The impressions were scanned, and reference points were selected on the occlusal surface of the contralateral first molars. MMF was calculated by subtracting the intermolar distance in O from the intermolar distance in R. Data were analysed by ANCOVA (fixed factors: facial pattern, sex; covariate: body mass index (BMI); alpha = 0.05). RESULTS: No significant difference of MOF or MMF was found among the three facial patterns (P = 0.62 and P = 0.72, respectively). BMI was not a significant covariate for MOF or MMF (P > 0.05). Sex was a significant factor only for MOF (P = 0.007); males had higher MOF values than females. CONCLUSION: These results suggest that MOF and MMF did not vary as a function of vertical facial pattern in this Brazilian sample. PMID- 17407567 TI - Respiratory symptoms among infants at risk for asthma: association with surfactant protein A haplotypes. AB - BACKGROUND: We examined the association between single nucleotide polymorphisms (SNPs) in loci encoding surfactant protein A (SFTPA) and risk of wheeze and persistent cough during the first year of life among a cohort of infants at risk for developing asthma. METHODS: Between September 1996 and December 1998, mothers of newborn infants were invited to participate if they had an older child with clinician-diagnosed asthma. Each mother was given a standardized questionnaire within 4 months of her infant's birth. Infant respiratory symptoms were collected during quarterly telephone interviews at 6, 9 and 12 months of age. Due to the association of SFTPA polymorphisms and race/ethnicity, analyses were restricted to 221 white infants for whom whole blood and respiratory data were available. Ordered logistic regression models were used to examine the association between respiratory symptom frequency and SFTPA haplotypes. RESULTS: The 6A allele haplotype of SFTPA1, with an estimated frequency of 6% among our study infants, was associated with an increased risk of persistent cough (OR 3.69, 95% CI 1.71, 7.98) and wheeze (OR 4.72, 95% CI 2.20, 10.11). The 6A/1A haplotype of SFTPA, found among approximately 5% of the infants, was associated with an increased risk of persistent cough (OR 3.20, 95% CI 1.39, 7.36) and wheeze (OR 3.25, 95% CI 1.43, 7.37). CONCLUSION: Polymorphisms within SFTPA loci may be associated with wheeze and persistent cough in white infants at risk for asthma. These associations require replication and exploration in other ethnic/racial groups. PMID- 17407568 TI - Mature habitats associated with genetic divergence despite strong dispersal ability in an arthropod. AB - BACKGROUND: Populations may be bound by contemporary gene flow, selective sweeps, and extinction-recolonization processes. Indeed, existing molecular estimates indicate that species with low levels of gene flow are rare. However, strong priority effects and local selective regimes may hinder gene flow (despite dispersal) sending populations on independent evolutionary trajectories. In this scenario (the monopolization hypothesis), population differentiation will increase with time and genealogical evidence should yield ample private haplotypes. Cyclical parthenogens (e.g. rotifers and cladocerans such as Daphnia) have an increased capacity for rapid local adaptation and priority effects because sexual reproduction is followed by multiple generations of clonal selection and massive egg bank formation. We aimed to better understand the history of population differentiation and ongoing gene flow in Daphnia rosea s.l., by comparing population and regional divergences in mature unglaciated areas and younger previously glaciated areas. We also examined the timing and paths of colonization of previously-glaciated areas to assess the dispersal limitations of D. rosea s.l. We used DNA sequence variation (84 populations and >400 individuals) at the mitochondrial ND2 and nuclear HSP90 loci from Holarctic populations for our genetic analyses. RESULTS: The genetic evidence indicated pronounced historical structure. Holarctic mtDNA phylogenies of D. rosea s.l. revealed three geographically restricted and divergent clades: European, Siberian and Japanese/American. The Japanese/American clade showed marked population genetic structure (FST > 0.8) that was weakly associated with geographic distance, and a high proportion of private haplotypes. Populations from older unglaciated habitats (i.e., Japan) showed higher DNA sequence divergences than populations from presumed younger habitats (i.e. non-Beringian North America) with nDNA and with mtDNA. Mismatch analyses of mtDNA and nDNA were consistent with a single rapid post-glacial expansion of D. rosea that covered most of the New World. CONCLUSION: Our evidence agrees with negligible gene flow after founding, and the accumulation of genetic divergence with habitat age. Existing direct evidence and our mismatch analyses indicate that the pronounced population differentiation is unlikely to be due to dispersal limitation. Instead, priority effects and local selection regimes may play a role in limiting gene flow. The results challenge the notion that lacustrine populations of cladocerans are generally unified by contemporary gene flow. PMID- 17407570 TI - Accuracy of self-reported height measurements in parents and its effect on mid parental target height calculation. AB - BACKGROUND: Clinical determination of mid-parental height is an important part of the assessment of a child's growth, however our clinical impression has been that parents cannot be relied upon to accurately report their own heights. Therefore, we conducted this study to assess the accuracy of parental height self-reporting and its effect on calculated mid-parental target height for children presenting to a pediatric endocrinology office. METHODS: All parents bringing their children for an initial evaluation to a pediatric endocrinology clinic over a period of nine months were questioned and then measured by a pediatric endocrinologist. Parents were blinded to the study. Mid-parental target heights, based on reported and actual height were compared. RESULTS: There were 241 families: 98 fathers and 217 mothers in our study. Mean measured paternal height was 173.2 cm, self reported 174.9 cm (p < 0.0001), partner reported 177 cm (p = 0.0004). Only 50% of fathers and 58% of mothers reported their height within +/- 2 cm of their measured height, while 15% of fathers and 12% of mothers were inaccurate by more than 4 cm. Mean measured maternal height was 160.6 cm, self-reported 161.1 cm (NS), partner reported 161.7 cm (NS). Inaccuracy of height self-report had a small but significant effect on the mean MPTH (0.4 cm, p = 0.045). Analysis showed that only 70% of MPTH calculated by reported heights fell within +/- 2 cm of MPTH calculated using measured heights, 24% being in +/- 2-4 cm range, and 6% were inaccurate by more than 4 cm. CONCLUSION: There is a significant difference in paternal measured versus reported heights with an overall trend for fathers to overestimate their own height. A large subset of parents makes a substantial error in their height self-report, which leads to erroneous MPTH. Inaccuracy is even greater when one parent reports the other parent's height. When a child's growth is in question, measured rather than reported parental heights should be obtained. PMID- 17407569 TI - The high-resolution NMR structure of the R21A Spc-SH3:P41 complex: understanding the determinants of binding affinity by comparison with Abl-SH3. AB - BACKGROUND: SH3 domains are small protein modules of 60-85 amino acids that bind to short proline-rich sequences with moderate-to-low affinity and specificity. Interactions with SH3 domains play a crucial role in regulation of many cellular processes (some are related to cancer and AIDS) and have thus been interesting targets in drug design. The decapeptide APSYSPPPPP (p41) binds with relatively high affinity to the SH3 domain of the Abl tyrosine kinase (Abl-SH3), while it has a 100 times lower affinity for the alpha-spectrin SH3 domain (Spc-SH3). RESULTS: Here we present the high-resolution structure of the complex between the R21A mutant of Spc-SH3 and p41 derived from NMR data. Thermodynamic parameters of binding of p41 to both WT and R21A Spc-SH3 were measured by a combination of isothermal titration and differential scanning calorimetry. Mutation of arginine 21 to alanine in Spc-SH3 increases 3- to 4-fold the binding affinity for p41 due to elimination at the binding-site interface of the steric clash produced by the longer arginine side chain. Amide hydrogen-deuterium experiments on the free and p41-bound R21A Spc-SH3 domain indicate that binding elicits a strong reduction in the conformational flexibility of the domain. Despite the great differences in the thermodynamic magnitudes of binding, the structure of the R21A Spc-SH3:P41 complex is remarkably similar to that of the Abl-SH3:P41 complex, with only few differences in protein-ligand contacts at the specificity pocket. Using empirical methods for the prediction of binding energetics based on solvent-accessible surface area calculations, the differences in experimental energetics of binding between the two complexes could not be properly explained only on the basis of the structural differences observed between the complexes. We suggest that the experimental differences in binding energetics can be at least partially ascribed to the absence in the R21A Spc-SH3:P41 complex of several buried water molecules, which have been proposed previously to contribute largely to the highly negative enthalpy and entropy of binding in the Abl-SH3:P41 complex. CONCLUSION: Based on a deep structural and thermodynamic analysis of a low and high affinity complex of two different SH3 domains with the same ligand p41, we underline the importance of taking into account in any effective strategy of rational design of ligands, factors different from the direct protein-ligand interactions, such as the mediation of interactions by water molecules or the existence of cooperative conformational effects induced by binding. PMID- 17407571 TI - A mitochondrial mutator plasmid that causes senescence under dietary restricted conditions. AB - BACKGROUND: Calorie or dietary restriction extends life span in a wide range of organisms including the filamentous fungus Podospora anserina. Under dietary restricted conditions, P. anserina isolates are several-fold longer lived. This is however not the case in isolates that carry one of the pAL2-1 homologous mitochondrial plasmids. RESULTS: We show that the pAL2-1 homologues act as 'insertional mutators' of the mitochondrial genome, which may explain their negative effect on life span extension. Sequencing revealed at least fourteen unique plasmid integration sites, of which twelve were located within the mitochondrial genome and two within copies of the plasmid itself. The plasmids were able to integrate in their entirety, via a non-homologous mode of recombination. Some of the integrated plasmid copies were truncated, which probably resulted from secondary, post-integrative, recombination processes. Integration sites were predominantly located within and surrounding the region containing the mitochondrial rDNA loci. CONCLUSION: We propose a model for the mechanism of integration, based on innate modes of mtDNA recombination, and discuss its possible link with the plasmid's negative effect on dietary restriction mediated life span extension. PMID- 17407572 TI - DNA microarray data integration by ortholog gene analysis reveals potential molecular mechanisms of estrogen-dependent growth of human uterine fibroids. AB - BACKGROUND: Uterine fibroids or leiomyoma are a common benign smooth muscle tumor. The tumor growth is well known to be estrogen-dependent. However, the molecular mechanisms of its estrogen-dependency is not well understood. METHODS: Differentially expressed genes in human uterine fibroids were either retrieved from published papers or from our own statistical analysis of downloaded array data. Probes for the same genes on different Affymetrix chips were mapped based on probe comparison information provided by Affymetrix. Genes identified by two or three array studies were submitted for ortholog analysis. Human and rat ortholog genes were identified by using ortholog gene databases, HomoloGene and TOGA and were confirmed by synteny analysis with MultiContigView tool in the Ensembl genome browser. RESULTS: By integrated analysis of three recently published DNA microarray studies with human tissue, thirty-eight genes were found to be differentially expressed in the same direction in fibroid compared to adjacent uterine myometrium by at least two research groups. Among these genes, twelve with rat orthologs were identified as estrogen-regulated from our array study investigating uterine expression in ovariectomized rats treated with estrogen. Functional and pathway analyses of the twelve genes suggested multiple molecular mechanisms for estrogen-dependent cell survival and tumor growth. Firstly, estrogen increased expression of the anti-apoptotic PCP4 gene and suppressed the expression of growth inhibitory receptors PTGER3 and TGFBR2. Secondly, estrogen may antagonize PPARgamma signaling, thought to inhibit fibroid growth and survival, at two points in the PPAR pathway: 1) through increased ANXA1 gene expression which can inhibit phospholipase A2 activity and in turn decrease arachidonic acid synthesis, and 2) by decreasing L-PGDS expression which would reduce synthesis of PGJ2, an endogenous ligand for PPARgamma. Lastly, estrogen affects retinoic acid (RA) synthesis and mobilization by regulating expression of CRABP2 and ALDH1A1. RA has been shown to play a significant role in the development of uterine fibroids in an animal model. CONCLUSION: Integrated analysis of multiple array datasets revealed twelve human and rat ortholog genes that were differentially expressed in human uterine fibroids and transcriptionally responsive to estrogen in the rat uterus. Functional and pathway analysis of these genes suggest multiple potential molecular mechanisms for the poorly understood estrogen-dependent growth of uterine fibroids. Fully understanding the exact molecular interactions among these gene products requires further study to validate their roles in uterine fibroids. This work provides new avenues of study which could influence the future direction of therapeutic intervention for the disease. PMID- 17407573 TI - Improved residue contact prediction using support vector machines and a large feature set. AB - BACKGROUND: Predicting protein residue-residue contacts is an important 2D prediction task. It is useful for ab initio structure prediction and understanding protein folding. In spite of steady progress over the past decade, contact prediction remains still largely unsolved. RESULTS: Here we develop a new contact map predictor (SVMcon) that uses support vector machines to predict medium- and long-range contacts. SVMcon integrates profiles, secondary structure, relative solvent accessibility, contact potentials, and other useful features. On the same test data set, SVMcon's accuracy is 4% higher than the latest version of the CMAPpro contact map predictor. SVMcon recently participated in the seventh edition of the Critical Assessment of Techniques for Protein Structure Prediction (CASP7) experiment and was evaluated along with seven other contact map predictors. SVMcon was ranked as one of the top predictors, yielding the second best coverage and accuracy for contacts with sequence separation > or = 12 on 13 de novo domains. CONCLUSION: We describe SVMcon, a new contact map predictor that uses SVMs and a large set of informative features. SVMcon yields good performance on medium- to long-range contact predictions and can be modularly incorporated into a structure prediction pipeline. PMID- 17407574 TI - Evidence for glycosylation on a DNA-binding protein of Salmonella enterica. AB - BACKGROUND: All organisms living under aerobic atmosphere have powerful mechanisms that confer their macromolecules protection against oxygen reactive species. Microorganisms have developed biomolecule-protecting systems in response to starvation and/or oxidative stress, such as DNA biocrystallization with Dps (DNA-binding protein from starved cells). Dps is a protein that is produced in large amounts when the bacterial cell faces harm, which results in DNA protection. In this work, we evaluated the glycosylation in the Dps extracted from Salmonella enterica serovar Typhimurium. This Dps was purified from the crude extract as an 18-kDa protein, by means of affinity chromatography on an immobilized jacalin column. RESULTS: The N-terminal sequencing of the jacalin bound protein revealed 100% identity with the Dps of S. enterica serovar Typhimurium. Methyl-alpha-galactopyranoside inhibited the binding of Dps to jacalin in an enzyme-linked lectin assay, suggesting that the carbohydrate recognition domain (CRD) of jacalin is involved in the interaction with Dps. Furthermore, monosaccharide compositional analysis showed that Dps contained mannose, glucose, and an unknown sugar residue. Finally, jacalin-binding Dps was detected in larger amounts during the bacterial earlier growth periods, whereas high detection of total Dps was verified throughout the bacterial growth period. CONCLUSION: Taken together, these results indicate that Dps undergoes post translational modifications in the pre- and early stationary phases of bacterial growth. There is also evidence that a small mannose-containing oligosaccharide is linked to this bacterial protein. PMID- 17407575 TI - Matrix-comparative genomic hybridization from multicenter formalin-fixed paraffin embedded colorectal cancer tissue blocks. AB - BACKGROUND: The identification of genomic signatures of colorectal cancer for risk stratification requires the study of large series of cancer patients with an extensive clinical follow-up. Multicentric clinical studies represent an ideal source of well documented archived material for this type of analyses. METHODS: To verify if this material is technically suitable to perform matrix-CGH, we performed a pilot study using macrodissected 29 formalin-fixed, paraffin-embedded tissue samples collected within the framework of the EORTC-GI/PETACC-2 trial for colorectal cancer. The scientific aim was to identify prognostic genomic signatures differentiating locally restricted (UICC stages II-III) from systemically advanced (UICC stage IV) colorectal tumours. RESULTS: The majority of archived tissue samples collected in the different centers was suitable to perform matrix-CGH. 5/7 advanced tumours displayed 13q-gain and 18q-loss. In locally restricted tumours, only 6/12 tumours showed a gain on 13q and 7/12 tumours showed a loss on 18q. Interphase-FISH and high-resolution array-mapping of the gain on 13q confirmed the validity of the array-data and narrowed the chromosomal interval containing potential oncogenes. CONCLUSION: Archival, paraffin-embedded tissue samples collected in multicentric clinical trials are suitable for matrix-CGH analyses and allow the identification of prognostic signatures and aberrations harbouring potential new oncogenes. PMID- 17407576 TI - Mismatch repair deficiencies transforming stem cells into cancer stem cells and therapeutic implications. AB - For the exceptional self-renewal capacity, regulated cell proliferation and differential potential to a wide variety of cell types, the stem cells must maintain the intact genome. The cells under continuous exogenous and endogenous genotoxic stress accumulate DNA errors, drive proliferative expansion and transform into cancer stem cells with a heterogeneous population of tumor cells. These cells are a common phenomenon for the hematological malignancies and solid tumors. In response to DNA damage, the complex cellular mechanisms including cell cycle arrest, transcription induction and DNA repair are activated. The cells when exposed to cytotoxic agents, the apoptosis lead to cell death. However, the absence of repair machinery makes the cells resistant to tumor sensitizing agents and result in malignant transformation. Mismatch repair gene defects are recently identified in hematopoietic malignancies, leukemia and lymphoma cell lines. This review emphasizes the importance of MMR systems in maintaining the stem cell functioning and its therapeutic implications in the eradication of cancer stem cells and differentiated tumor cells as well. The understanding of the biological functions of mismatch repair in the stem cells and its malignant counterparts could help in developing an effective novel therapies leaving residual non tumorigenic population of cells resulting in potential cancer cures. PMID- 17407577 TI - Zerumbone induced apoptosis in liver cancer cells via modulation of Bax/Bcl-2 ratio. AB - BACKGROUND: Zerumbone is a cytotoxic component isolated from Zingiber zerumbet Smith, a herbal plant which is also known as lempoyang. This new anticancer bioactive compound from Z. zerumbet was investigated for its activity and mechanism in human liver cancer cell lines. RESULTS: Zerumbone significantly showed an antiproliferative activity upon HepG2 cells with an IC50 of 3.45 +/- 0.026 microg/ml. Zerumbone was also found to inhibit the proliferation of non malignant Chang Liver and MDBK cell lines. However the IC50 obtained was higher compared to the IC50 for HepG2 cells (> 10 microg/ml). The extent of DNA fragmentation was evaluated by the Tdt-mediated dUTP nick end labelling assay which showed that, zerumbone significantly increased apoptosis in HepG2 cells in a time-course manner. In detail, the apoptotic process triggered by zerumbone involved the up-regulation of pro-apoptotic Bax protein and the suppression of anti-apoptotic Bcl-2 protein expression. The changes that occurred in the levels of this antagonistic proteins Bax/Bcl-2, was independent of p53 since zerumbone did not affect the levels of p53 although this protein exists in a functional form. Western blotting analysis for Bax protein was further confirmed qualitatively with an immunoassay that showed the distribution of Bax protein in zerumbone-treated cells. CONCLUSION: Therefore, zerumbone was found to induce the apoptotic process in HepG2 cells through the up and down regulation of Bax/Bcl-2 protein independently of functional p53 activity. PMID- 17407578 TI - Characteristics of beta-lactamases and their genes (blaA and blaB) in Yersinia intermedia and Y. frederiksenii. AB - BACKGROUND: The presence of beta-lactamases in Y. enterocolitica has been reported to vary with serovars, biovars and geographical origin of the isolates. An understanding of the beta-lactamases in other related species is important for an overall perception of antibiotic resistance in yersiniae. The objective of this work was to study the characteristics of beta-lactamases and their genes in strains of Y. intermedia and Y. frederiksenii, isolated from clinical and non clinical sources in India. RESULTS: The enzymes, Bla-A (a constitutive class A penicillinase) and Bla-B (an inducible class C cephalosporinase) were found to be present in all the clinical and non-clinical strains of Y. intermedia and Y. frederiksenii by double disc diffusion method. The results showed differential expression of Bla-A as indicated by presence/absence of synergy whereas expression of Bla-B was quite consistent. The presence of these enzymes was also reflected in the high minimum inhibitory concentrations, MIC50 (126-1024 mg/L) and MIC90 (256-1024 mg/L) of beta-lactam antibiotics against these species. Restriction fragment length polymorphism (RFLP) revealed heterogeneity in both blaA and blaB genes of Y. intermedia and Y. frederiksenii. The blaA gene of Y. intermedia shared significant sequence identity (87-96%) with blaA of Y. enterocolitica biovars 1A, 1B and 4. The sequence identity of blaA of Y. frederiksenii with these biovars was 77-79%. The sequence identity of blaB gene of Y. intermedia and Y. frederiksenii was more (85%) with that of Y. enterocolitica biovars 1A, 1B and 2 compared to other species viz., Y. bercovieri, Y. aldovae and Y. ruckeri. Isoelectric focusing data further revealed that both Y. intermedia and Y. frederiksenii produced Bla-A (pI 8.7) and "Bla-B like" (pI 5.5-7.1) enzymes. CONCLUSION: Both Y. intermedia and Y. frederiksenii showed presence of blaA and blaB genes and unequivocal expression of the two beta lactamases. Limited heterogeneity was detected in blaA and blaB genes as judged by PCR-RFLP. Phylogenetic relationships showed that the two species shared a high degree of identity in their bla genes. This is the first study reporting characteristics of beta-lactamases and their genes in strains of Y. intermedia and Y. frederiksenii isolated from Asian region. PMID- 17407579 TI - Cross-species analysis of gene expression in non-model mammals: reproducibility of hybridization on high density oligonucleotide microarrays. AB - BACKGROUND: Gene expression profiles of non-model mammals may provide valuable data for biomedical and evolutionary studies. However, due to lack of sequence information of other species, DNA microarrays are currently restricted to humans and a few model species. This limitation may be overcome by using arrays developed for a given species to analyse gene expression in a related one, an approach known as "cross-species analysis". In spite of its potential usefulness, the accuracy and reproducibility of the gene expression measures obtained in this way are still open to doubt. The present study examines whether or not hybridization values from cross-species analyses are as reproducible as those from same-species analyses when using Affymetrix oligonucleotide microarrays. RESULTS: The reproducibility of the probe data obtained hybridizing deer, Old World primates, and human RNA samples to Affymetrix human GeneChip U133 Plus 2.0 was compared. The results show that cross-species hybridization affected neither the distribution of the hybridization reproducibility among different categories, nor the reproducibility values of the individual probes. Our analyses also show that a 0.5% of the probes analysed in the U133 plus 2.0 GeneChip are significantly associated to un-reproducible hybridizations. Such probes-called in the text un-reproducible probe sequences- do not increase in number in cross species analyses. CONCLUSION: Our study demonstrates that cross-species analyses do not significantly affect hybridization reproducibility of GeneChips, at least within the range of the mammal species analysed here. The differences in reproducibility between same-species and cross-species analyses observed in previous studies were probably caused by the analytical methods used to calculate the gene expression measures. Together with previous observations on the accuracy of GeneChips for cross-species analysis, our analyses demonstrate that cross species hybridizations may provide useful gene expression data. However, the reproducibility and accuracy of these measures largely depends on the use of appropriated algorithms to derive the gene expression data from the probe data. Also, the identification of probes associated to un-reproducible hybridizations useless for gene expression analyses- in the studied GeneChip, stress the need of a re-evaluation of the probes' performance. PMID- 17407580 TI - The reliability of a 10-test package for patients with prolonged back and neck pain: could an examiner without formal medical education be used without loss of quality? A methodological study. AB - BACKGROUND: In the rehabilitation of patients with prolonged back and neck pain, the physical impairment should be assessed. Previous research has exclusively engaged medically educated examiners, mostly physiotherapists. However, less biased evaluations of efforts at rehabilitation might be achieved by personnel standing outside the treatment work itself. Therefore, if medically untrained examiners could be used without cost to the quality, this might produce a better evaluation at defensible cost and could also be useful in a research context. The aim of this study was to answer the question: given a 10-test package for patients with prolonged back and neck pain, could an examiner without formal medical education be used without loss of quality? Five of the ten tests required the examiner to keep a firm hold against the foundation of those parts of the participant's body that were not supposed to move during the test. METHODS: Examination by an experienced physiotherapist (A) in performing the package was compared with that by a research assistant (B) without formal medical education. The reliability, including inter- and intra-rater reliability, was assessed. In the inter-rater reliability study, 50 participants (30 patients + 20 healthy subjects) were tested once each by A and B. In the intra-rater reliability study, the 20 healthy subjects were tested twice by A or B. One-way ANOVA intra-class correlation coefficient (ICC) was calculated and its possible systematic error was determined using a t-test. RESULTS: All five tests that required no manual fixation had acceptable reliability (ICC > .60 and no indication of systematic error). Only one of the five tests that required fixation had acceptable reliability. The difference (five vs. one) was significant (p = .01). CONCLUSION: In a 10-test package for patients with prolonged back and neck pain, an examiner without formal medical education could be used without loss of quality, at least for the five tests requiring no manual fixation. To make our results more generalizable and their implications more searching, a similar study should be conducted with two or more examiners with and without formal medical education, and the intra-rater reliability study should also include patients and involve more participants. PMID- 17407581 TI - Are passive smoking, air pollution and obesity a greater mortality risk than major radiation incidents? AB - BACKGROUND: Following a nuclear incident, the communication and perception of radiation risk becomes a (perhaps the) major public health issue. In response to such incidents it is therefore crucial to communicate radiation health risks in the context of other more common environmental and lifestyle risk factors. This study compares the risk of mortality from past radiation exposures (to people who survived the Hiroshima and Nagasaki atomic bombs and those exposed after the Chernobyl accident) with risks arising from air pollution, obesity and passive and active smoking. METHODS: A comparative assessment of mortality risks from ionising radiation was carried out by estimating radiation risks for realistic exposure scenarios and assessing those risks in comparison with risks from air pollution, obesity and passive and active smoking. RESULTS: The mortality risk to populations exposed to radiation from the Chernobyl accident may be no higher than that for other more common risk factors such as air pollution or passive smoking. Radiation exposures experienced by the most exposed group of survivors of Hiroshima and Nagasaki led to an average loss of life expectancy significantly lower than that caused by severe obesity or active smoking. CONCLUSION: Population-averaged risks from exposures following major radiation incidents are clearly significant, but may be no greater than those from other much more common environmental and lifestyle factors. This comparative analysis, whilst highlighting inevitable uncertainties in risk quantification and comparison, helps place the potential consequences of radiation exposures in the context of other public health risks. PMID- 17407582 TI - Food safety in hospital: knowledge, attitudes and practices of nursing staff of two hospitals in Sicily, Italy. AB - BACKGROUND: Food hygiene in hospital poses peculiar problems, particularly given the presence of patients who could be more vulnerable than healthy subjects to microbiological and nutritional risks. Moreover, in nosocomial outbreaks of infectious intestinal disease, the mortality risk has been proved to be significantly higher than the community outbreaks and highest for foodborne outbreaks. On the other hand, the common involvement in the role of food handlers of nurses or domestic staff, not specifically trained about food hygiene and HACCP, may represent a further cause of concern. The purpose of this study was to evaluate knowledge, attitudes, and practices concerning food safety of the nursing staff of two hospitals in Palermo, Italy. Association with some demographic and work-related determinants was also investigated. METHODS: The survey was conducted, by using a semi-structured questionnaire, in March-November 2005 in an acute general hospital and a paediatric hospital, where nursing staff is routinely involved in food service functions. RESULTS: Overall, 401 nurses (279, 37.1%, of the General Hospital and 122, 53.5%, of the Paediatric Hospital, respectively) answered. Among the respondents there was a generalized lack of knowledge about etiologic agents and food vehicles associated to foodborne diseases and proper temperatures of storage of hot and cold ready to eat foods. A general positive attitude towards temperature control and using clothing and gloves, when handling food, was shared by the respondents nurses, but questions about cross-contamination, refreezing and handling unwrapped food with cuts or abrasions on hands were frequently answered incorrectly. The practice section performed better, though sharing of utensils for raw and uncooked foods and thawing of frozen foods at room temperatures proved to be widely frequent among the respondents. Age, gender, educational level and length of service were inconsistently associated with the answer pattern. More than 80% of the respondent nurses did not attend any educational course on food hygiene. Those who attended at least one training course fared significantly better about some knowledge issues, but no difference was detected in both the attitude and practice sections. CONCLUSION: Results strongly emphasize the need for a safer management of catering in the hospitals, where non professional food handlers, like nursing or domestic staff, are involved in food service functions. PMID- 17407583 TI - Factors associated with dog ownership and contact with dogs in a UK community. AB - BACKGROUND: Dogs are popular pets in many countries. Identifying differences between those who own dogs or have contact with dogs, and those who do not, is useful to those interested in the human-animal bond, human health and for provision of veterinary services. This census-based, epidemiological study aimed to investigate factors associated with dog ownership and contact with dogs, in a semi-rural community of 1278 households in Cheshire, UK. RESULTS: Twenty-four percent of households were identified as dog-owning and 52% owned a pet of some type. Multivariable logistic regression suggested that households were more likely to own a dog if they had more occupants (five or more); if they had an adult female household member; or if they owned a horse. The age structure of the households was also associated with dog ownership, with households containing older children (between six and 19 years of age) and young adults (between 20 and 29 years of age), more likely to own dogs. We also found that dog owning households were more likely to be multi-dog households than single-dog if they also owned a cat or a bird, or if the household contained a person of 20-29 years old. Dog owners reported increased contact with dogs, other than their own, compared to those that did not own dogs and this contact appeared to be mainly through walking. CONCLUSION: Some household types are more likely to own a dog than others. This study supports the suggestion that dogs are more common in families who have older children (6-19 years), as has been generally observed in other countries. Dog owners are also more likely to have contact with dogs other than their own, compared with those not owning a dog. PMID- 17407584 TI - Rapamycin-induced inhibition of HTLV-I LTR activity is rescued by c-Myb. AB - BACKGROUND: Rapamycin is an immunosuppressive which represses translation of transcripts harbouring a polypyrimidine motif downstream of the mRNA cap site through the mammalian target of rapamycin complex. It inhibits the abnormal autologous proliferation of T-cell clones containing a transcriptionally active human T-lymphotropic virus, type I (HTLV-I) provirus, generated from infected subjects. We showed previously that this effect is independent of the polypyrimidine motifs in the viral long terminal repeat (LTR) R region suggesting that HTLV-I transcription, and not translation, is being affected. Here we studied whether rapamycin is having an effect on a specific transcription factor pathway. Further, we investigated whether mRNAs encoding transcription factors involved in HTLV-I transcriptional activation, specifically CREB, Ets and c-Myb, are implicated in the rapamycin-sensitivity of the HTLV-I LTR. RESULTS: An in vitro analysis of the role of SRE- and NF-kappaB-mediated transcription highlighted the latter as rapamycin sensitive. Over-expression of c-Myb reversed the rapamycin effect. CONCLUSION: The sensitivity of HTLV-I transcription to rapamycin may be effected through an NF-kappaB-pathway associated with the rapamycin-sensitive mTORC1 cellular signalling network. PMID- 17407585 TI - Microdecompression for lumbar synovial cysts: an independent assessment of long term outcomes. AB - BACKGROUND: Outcomes of surgical intervention for lumbar synovial cysts have been evaluated in the short and intermediate term. Concerns regarding cyst recurrence, the development of late instability at the involved level, and instability/stenosis at adjacent levels (when concomitant) fusion is performed suggest that long term follow-up is needed. This study aims to fill that void. METHODS: Forty-six patients operated by a single surgeon not involved in the study were followed up long term at an average of 9.7 years (range 5 to 22 years) post-operatively. All patients underwent decompression (+/- concomitant arthrodesis in the presence of associated degenerative spondylolisthesis) using the operative microscope for magnification/illumination. Outcomes were assessed using a customized questionnaire evaluating: relief of pain/claudicant symptoms, numbness/parasthesias, and weakness; as well as late onset low back pain, new radicular symptoms, need for additional surgery, and patient satisfaction. Outcomes in patients with or without fusion were compared as well. RESULTS: 87% of patients noted resolution of their pre-operative pain, numbness, and weakness. 28% of patients developed late onset low back pain. 17% developed late onset radicular symptoms in a new nerve root distribution. 15% required subsequent additional surgery. 89% of patients were satisfied with the surgical outcome. No differences were found for any outcome measure between patients undergoing concomitant fusion and those undergoing decompression alone using the two-sample t-test. CONCLUSION: This study provides outcome data at an average of nearly ten years post-operative. This information should allow surgeons to provide realistic expectations for their patients regarding outcomes and should enhance the informed consent and surgical decision-making process. PMID- 17407586 TI - Some facts and thoughts: p73 as a tumor suppressor gene in the network of tumor suppressors. AB - The question of whether p73 is a tumor suppressor gene, is not yet answered with full confidence. The lack of spontaneous tumor formation in p73 null mice and infrequent p73 mutations seen in a variety of cancers analyzed would straightaway negate its role as a primary tumor suppressor gene. However, accumulating evidence suggest that p73 gene and its target genes are hypermethylated in the cancer of lymphoid origin. Here I discuss some facts and thoughts that support the idea that p73 could still be a tumor suppressor gene. The tumor suppressor network in which p73 appears to be a participant involves E2F1, JunB, INK4a/p16, ARF/p19, p57kip2 and BRCA1. Knock out of each gene in E2F-1-p73-JunB-p16INK4a network of tumor suppressor proteins result in lymphoma/leukemia formation. Further, I tried to explain why lymphomas are not seen in p73 null mice and why p73 gene is not prone to frequent mutation. PMID- 17407587 TI - Irbesartan for the treatment of hypertension in patients with the metabolic syndrome: a sub analysis of the Treat to Target post authorization survey. Prospective observational, two armed study in 14,200 patients. AB - OBJECTIVES: The metabolic syndrome is a cluster of cardiovascular risk factors leading to an increased risk for the subsequent development of diabetes and cardiovascular morbidity and mortality. Blocking the renin-angiotensin system has been shown to prevent cardiovascular disease and delay the onset of diabetes. Irbesartan is an angiotensin receptor blocker (ARB) which has been shown to possess peroxisome proliferator-activated receptor gamma (PPARgamma) activating properties, and to have a favorable metabolic profile. Current discussion is whether the addition of small doses of hydrochlorothiazide changes this profile. Therefore the efficacy, safety and metabolic profile of Irbesartan either as monotherapy or in combination therapy was assessed in patients with the metabolic syndrome in a large observational cohort in primary care. RESEARCH DESIGN AND METHODS: Multicenter, prospective, two-armed, post authorization study over 9 months in 14,200 patients with uncontrolled hypertension with and without the metabolic syndrome (doctors' diagnosis based on the Adult Treatment Panel III criteria 2001). Blood pressure was measured sphygmomanometrically and cardiovascular risk factors making up the criteria for the metabolic syndrome were assessed. MAIN OUTCOME MEASURES: Systolic (SBP) and diastolic (DBP) blood pressure reduction, response, and normalization (systolic and diastolic), changes in fasting glucose, waist circumference (abdominal obesity), serum triglycerides and HDL cholesterol as well as the proportion of patients fulfilling the criteria for the metabolic syndrome. Number and nature of adverse events (AEs). RESULTS: After 9 month the use of Irbesartan in monotherapy resulted in a significant reduction of blood pressure (SBP: -26.3 +/- 10.1 mmHg/DBP-13.0 +/- 6.6 mmHg, both p < 0.0001) in patients with the metabolic syndrome. This was accompanied by a reduction in cardiovascular risk factors: HDL cholesterol (+3.6 +/- 7.2 mg/dl in men, +3.8 +/- 6.5 mg/dl in women, both p < 0.0001), serum triglycerides (-28.6 +/ 52.1 mg/dl, p < 0.0001), fasting blood glucose (-8.4 +/- 25.1 mg/dl, p < 0.0001) and waist circumference (-2.4 +/- 11.9 cm in men, -1.2 +/- 14.2 in women, both p < 0.0001) were significantly improved. Irbesartan combination therapy (12.5 mg HCTZ) in patients with the metabolic syndrome: blood pressure reduction (SBP: 27.5 +/- 10.1 mmHg/DBP: -14.1 +/- 6.6 mmHg, both p < 0.0001), improvement in HDL cholesterol (+4.0 +/- 6.8 mg/dl in men, +3.4 +/- 6.8 in women, both p < 0.0001), triglycerides (-34.1 +/- 52.6 mg/dl, p < 0.0001), fasting blood glucose (-10.0 +/ 24.7, p < 0.0001) and waist circumference (-3.2 +/- 12.7 cm in men, -1.7 +/- 14.4 in women, both p < 0.0001). Tolerability was excellent: only 0.6% of patients experienced an AE. CONCLUSION: There was a significant improvement in blood pressure and metabolic risk factors as a result of Irbesartan treatment. There was no evidence of a difference between monotherapy and combination therapy with regard to the cardiovascular risk profile. PMID- 17407592 TI - CAMbase--a XML-based bibliographical database on Complementary and Alternative Medicine (CAM). AB - The term "Complementary and Alternative Medicine (CAM)" covers a variety of approaches to medical theory and practice, which are not commonly accepted by representatives of conventional medicine. In the past two decades, these approaches have been studied in various areas of medicine. Although there appears to be a growing number of scientific publications on CAM, the complete spectrum of complementary therapies still requires more information about published evidence. A majority of these research publications are still not listed in electronic bibliographical databases such as MEDLINE. However, with a growing demand by patients for such therapies, physicians increasingly need an overview of scientific publications on CAM. Bearing this in mind, CAMbase, a bibliographical database on CAM was launched in order to close this gap. It can be accessed online free of charge or additional costs. The user can peruse more than 80,000 records from over 30 journals and periodicals on CAM, which are stored in CAMbase. A special search engine performing syntactical and semantical analysis of textual phrases allows the user quickly to find relevant bibliographical information on CAM. Between August 2003 and July 2006, 43,299 search queries, an average of 38 search queries per day, were registered focussing on CAM topics such as acupuncture, cancer or general safety aspects. Analysis of the requests led to the conclusion that CAMbase is not only used by scientists and researchers but also by physicians and patients who want to find out more about CAM. Closely related to this effort is our aim to establish a modern library center on Complementary Medicine which offers the complete spectrum of a modern digital library including a document delivery-service for physicians, therapists, scientists and researchers. PMID- 17407591 TI - Palliative care provision for patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a major cause of disability, morbidity and mortality in old age. Patients with advanced stage COPD are most likely to be admitted three to four times per year with acute exacerbations of COPD (AECOPD) which are costly to manage. The adverse events of AECOPD are associated with poor quality of life, severe physical disability, loneliness, and depression and anxiety symptoms. Currently there is a lack of palliative care provision for patients with advanced stage COPD compared with cancer patients despite having poor prognosis, intolerable dyspnoea, lower levels of self efficacy, greater disability, poor quality of life and higher levels of anxiety and depression. These symptoms affect patients' quality of life and can be a source of concern for family and carers as most patients are likely to be housebound and may be in need of continuous support and care. Evidence of palliative care provision for cancer patients indicate that it improves quality of life and reduces health care costs. The reasons why COPD patients do not receive palliative care are complex. This partly may relate to prognostic accuracy of patients' survival which poses a challenge for healthcare professionals, including general practitioners for patients with advanced stage COPD, as they are less likely to engage in end-of-life care planning in contrast with terminal disease like cancer. Furthermore there is a lack of resources which constraints for the wider availability of the palliative care programmes in the health care system. Potential barriers may include unwillingness of patients to discuss advance care planning and end-of-life care with their general practitioners, lack of time, increased workload, and fear of uncertainty of the information to provide about the prognosis of the disease and also lack of appropriate tools to guide general practitioners when to refer patients for palliative care. COPD is a chronic incurable disease; those in an advanced stage of the disease pursuing intensive medical treatment may also benefit from the simultaneous holistic care approach of palliative care services, medical services and social services to improve quality of end of life care. PMID- 17407589 TI - Deafblindness in French Canadians from Quebec: a predominant founder mutation in the USH1C gene provides the first genetic link with the Acadian population. AB - BACKGROUND: Usher syndrome type 1 (USH1) is the leading cause of deafblindness. In most populations, many private mutations are distributed across the five known USH1 genes. We investigated patients from the French Canadian population of Quebec (approximately 6 million people) that descends from about 8,500 French settlers who colonized the St Lawrence River valley between 1608 and 1759. We hypothesized that founder mutations in USH1 genes exist in this population. RESULTS: We have genetically characterized 15 patients from different regions of Quebec who were clinically diagnosed as USH1. Of these cases, 60% carried mutations of the USH1C gene, a genetic subtype that is rare outside the Acadian population. We have discovered a founder effect of the c.216G>A mutation, which has previously been designated the 'Acadian allele' because it accounts for virtually all Acadian USH1 cases. It represents 40% of disease alleles in Quebec, and a carrier of c.216G>A was identified in the general population. Mutations in other genes, except CDH23, are very rare. CONCLUSION: Based on our findings, approximately 0.5% of congenitally deaf children in Quebec are at risk of developing retinal degeneration due to homozygosity for c.216G>A. Although the Acadians and French Canadians from Quebec are descended from French ancestors, they have always been considered genetically distinct. The genetic conditions common in Quebec are generally not found in Acadians, or they are due to different mutations. Our results, however, show that carriers of the c.216G>A allele haplotype belonged to the early founders of both the Acadian and the Quebec population. PMID- 17407588 TI - Squamous cell carcinoma of the prostate: long-term survival after combined chemo radiation. AB - BACKGROUND: Carcinoma of the prostate gland is the most frequent malignant tumour affecting male population. While the large majority of tumours is represented by adenocarcinoma, pure squamous cell carcinoma comprises only 0.5-1% of all prostate neoplastic lesions. It is characterised by a high degree of malignancy, commonly metastasising to the bone (mainly with osteolytic lesions), liver and lungs with a median survival time of 14 months. Several therapeutic approaches have been employed in the effort to treat prostate pure squamous cell carcinoma, including radical surgery, radiotherapy, chemotherapy and hormonal therapy. All of them mostly failed to gain a significant survival benefit. CASE REPORT: We herein report on a case of pure squamous cell carcinoma of the prostate approached with combined-modality treatment, with the administration of 3 courses of cisplatin 75 mg/m(2) on day 1 and continuous infusion 5-fluorouracil 750 mg/m2 on day 1 to 5 and, subsequently, radiotherapy, with the delivery of a total dose of 46 Gy to the whole pelvis, with additional boost doses of 20 Gy to the prostatic bed and adjunctive 6 Gy to the prostate gland (72 Gy in total). The patient remained free of disease for 5 years, finally experiencing local relapse and, subsequently, dying of acute renal failure due to bilateral uretero-hydro nephrosis. In addition, we provide a complete overview of all reported cases available within the medical literature. CONCLUSION: Since it remains questionable which should be the most appropriate therapeutic approach towards prostate pure squamous cell carcinoma, our report demonstrates that a prolonged disease control, with a consistent survival time, may be achieved by the combination of an effective local treatment such as radiotherapy with systemic infusion of chemotherapeutic drugs. PMID- 17407590 TI - Understanding LTP in pain pathways. AB - Long-term potentiation (LTP) at synapses of nociceptive nerve fibres is a proposed cellular mechanism underlying some forms of hyperalgesia. In this review fundamental properties of LTP in nociceptive pathways are described. The following topics are specifically addressed: A concise definition of LTP is given and a differentiation is made between LTP and "central sensitisation". How to (and how not to) measure and how to induce LTP in pain pathways is specified. The signal transduction pathways leading to LTP at C-fibre synapses are highlighted and means of how to preempt and how to reverse LTP are delineated. The potential functional roles of LTP are evaluated at the cellular level and at the behavioural level in experimental animals. Finally, the impact of LTP on the perception of pain in human subjects is discussed. PMID- 17407593 TI - Molecular genetics of nicotine dependence and abstinence: whole genome association using 520,000 SNPs. AB - BACKGROUND: Classical genetic studies indicate that nicotine dependence is a substantially heritable complex disorder. Genetic vulnerabilities to nicotine dependence largely overlap with genetic vulnerabilities to dependence on other addictive substances. Successful abstinence from nicotine displays substantial heritable components as well. Some of the heritability for the ability to quit smoking appears to overlap with the genetics of nicotine dependence and some does not. We now report genome wide association studies of nicotine dependent individuals who were successful in abstaining from cigarette smoking, nicotine dependent individuals who were not successful in abstaining and ethnically matched control subjects free from substantial lifetime use of any addictive substance. RESULTS: These data, and their comparison with data that we have previously obtained from comparisons of four other substance dependent vs control samples support two main ideas: 1) Single nucleotide polymorphisms (SNPs) whose allele frequencies distinguish nicotine-dependent from control individuals identify a set of genes that overlaps significantly with the set of genes that contain markers whose allelic frequencies distinguish the four other substance dependent vs control groups (p < 0.018). 2) SNPs whose allelic frequencies distinguish successful vs unsuccessful abstainers cluster in small genomic regions in ways that are highly unlikely to be due to chance (Monte Carlo p < 0.00001). CONCLUSION: These clustered SNPs nominate candidate genes for successful abstinence from smoking that are implicated in interesting functions: cell adhesion, enzymes, transcriptional regulators, neurotransmitters and receptors and regulation of DNA, RNA and proteins. As these observations are replicated, they will provide an increasingly-strong basis for understanding mechanisms of successful abstinence, for identifying individuals more or less likely to succeed in smoking cessation efforts and for tailoring therapies so that genotypes can help match smokers with the treatments that are most likely to benefit them. PMID- 17407594 TI - relocating job wise? A mathematical model separates quantitatively the cytostatic and cytotoxic effects of a HER2 tyrosine kinase inhibitor. AB - BACKGROUND: Oncogene signaling is known to deregulate cell proliferation resulting in uncontrolled growth and cellular transformation. Gene amplification and/or somatic mutations of the HER2/Neu (ErbB2) proto-oncogene occur in approximately 20% of breast cancers. A therapeutic strategy that has been used to block HER2 function is the small molecule tyrosine kinase inhibitor lapatinib. Using human mammary epithelial cells that overexpress HER2, we determined the anti-proliferative effect of lapatinib through measuring the total cell number and analyzing the cell cycle distribution. A mathematical model was used to interpret the experimental data. RESULTS: The model suggests that lapatinib acts as expected by slowing the transition through G1 phase. However, the experimental data indicated a previously unreported late cytotoxic effect, which was incorporated into the model. Both effects depend on the dosage of the drug, which shows saturation kinetics. CONCLUSION: The model separates quantitatively the cytostatic and cytotoxic effects of lapatinib and may have implications for preclinical studies with other anti-oncogene therapies. PMID- 17407595 TI - Neither MRI, CT nor US is superior to diagnose tumors in the salivary glands--an extended case study. AB - OBJECTIVES: Ultrasonography (US), computed tomography (CT) and magnetic resonance imaging (MRI) are the most common radiological procedures for the diagnosis of tumor-like lesions of the salivary glands. The aim of the present study was to determine whether MRI or CT provide additional information besides that delivered by US. STUDY DESIGN/METHODS: 109 patients with a tumor-like lesion of the salivary glands underwent surgery. MRI and CT were arranged in 73 and in 40 patients respectively, whereas all 109 patients were prospectively diagnosed by US. The results of CT, MRI and US were compared with the histological outcome. Furthermore, the recent rise in the number of CT and MRI studies was investigated. RESULTS: On CT and MRI, there was no rise in the percentage of malignant tumors or advanced surgical procedures. In respect of the radiological assessment of the lesion (benign/malignant) and the correct diagnosis, CT, MRI and US were comparable in terms of sensitivity, specificity and accuracy. No significant difference was found in the Chi-square test (p > 0.05). CONCLUSION: The evaluation of the preoperative results of CT, MRI and US revealed no advantage for CT or MRI; these procedures are only required in specific cases. An update or revision of the current preoperative diagnostic management is deemed necessary. PMID- 17407596 TI - From genes to functional classes in the study of biological systems. AB - BACKGROUND: With the popularization of high-throughput techniques, the need for procedures that help in the biological interpretation of results has increased enormously. Recently, new procedures inspired in systems biology criteria have started to be developed. RESULTS: Here we present FatiScan, a web-based program which implements a threshold-independent test for the functional interpretation of large-scale experiments that does not depend on the pre-selection of genes based on the multiple application of independent tests to each gene. The test implemented aims to directly test the behaviour of blocks of functionally related genes, instead of focusing on single genes. In addition, the test does not depend on the type of the data used for obtaining significance values, and consequently different types of biologically informative terms (gene ontology, pathways, functional motifs, transcription factor binding sites or regulatory sites from CisRed) can be applied to different classes of genome-scale studies. We exemplify its application in microarray gene expression, evolution and interactomics. CONCLUSION: Methods for gene set enrichment which, in addition, are independent from the original data and experimental design constitute a promising alternative for the functional profiling of genome-scale experiments. A web server that performs the test described and other similar ones can be found at: http://www.babelomics.org. PMID- 17407597 TI - De novo identification of LTR retrotransposons in eukaryotic genomes. AB - BACKGROUND: LTR retrotransposons are a class of mobile genetic elements containing two similar long terminal repeats (LTRs). Currently, LTR retrotransposons are annotated in eukaryotic genomes mainly through the conventional homology searching approach. Hence, it is limited to annotating known elements. RESULTS: In this paper, we report a de novo computational method that can identify new LTR retrotransposons without relying on a library of known elements. Specifically, our method identifies intact LTR retrotransposons by using an approximate string matching technique and protein domain analysis. In addition, it identifies partially deleted or solo LTRs using profile Hidden Markov Models (pHMMs). As a result, this method can de novo identify all types of LTR retrotransposons. We tested this method on the two pairs of eukaryotic genomes, C. elegans vs. C. briggsae and D. melanogaster vs. D. pseudoobscura. LTR retrotransposons in C. elegans and D. melanogaster have been intensively studied using conventional annotation methods. Comparing with previous work, we identified new intact LTR retroelements and new putative families, which may imply that there may still be new retroelements that are left to be discovered even in well-studied organisms. To assess the sensitivity and accuracy of our method, we compared our results with a previously published method, LTR_STRUC, which predominantly identifies full-length LTR retrotransposons. In summary, both methods identified comparable number of intact LTR retroelements. But our method can identify nearly all known elements in C. elegans, while LTR_STRUCT missed about 1/3 of them. Our method also identified more known LTR retroelements than LTR_STRUCT in the D. melanogaster genome. We also identified some LTR retroelements in the other two genomes, C. briggsae and D. pseudoobscura, which have not been completely finished. In contrast, the conventional method failed to identify those elements. Finally, the phylogenetic and chromosomal distributions of the identified elements are discussed. CONCLUSION: We report a novel method for de novo identification of LTR retrotransposons in eukaryotic genomes with favorable performance over the existing methods. PMID- 17407598 TI - Comparative proteomic analysis of cell lines and scrapings of the human intestinal epithelium. AB - BACKGROUND: In vitro models are indispensable study objects in the fields of cell and molecular biology, with advantages such as accessibility, homogeneity of the cell population, reproducibility, and growth rate. The Caco-2 cell line, originating from a colon carcinoma, is a widely used in vitro model for small intestinal epithelium. Cancer cells have an altered metabolism, making it difficult to infer their representativity for the tissue from which they are derived. This study was designed to compare the protein expression pattern of Caco-2 cells with the patterns of intestinal epithelial cells from human small and large intestine. HT-29 intestinal cells, Hep G2 liver cells and TE 671 muscle cells were included too, the latter two as negative controls. RESULTS: Two dimensional gel electrophoresis was performed on each tissue and cell line protein sample. Principal component and cluster analysis revealed that global expression of intestinal epithelial scrapings differed from that of intestinal epithelial cell lines. Since all cultured cell lines clustered together, this finding was ascribed to an adaptation of cells to culture conditions and their tumor origin, and responsible proteins were identified by mass spectrometry. When investigating the profiles of Caco-2 cells and small intestinal cells in detail, a considerable overlap was observed. CONCLUSION: Numerous proteins showed a similar expression in Caco-2 cells, HT-29 cells, and both the intestinal scrapings, of which some appear to be characteristic to human intestinal epithelium in vivo. In addition, several biologically significant proteins are expressed at comparable levels in Caco-2 cells and small intestinal scrapings, indicating the usability of this in vitro model. Caco-2 cells, however, appear to over-express as well as under-express certain proteins, which needs to be considered by scientists using this cell line. Hence, care should be taken to prevent misinterpretation of in vitro obtained findings when translating them to the in vivo situation. PMID- 17407599 TI - A recessive genetic screen for host factors required for retroviral infection in a library of insertionally mutated Blm-deficient embryonic stem cells. AB - BACKGROUND: Host factors required for retroviral infection are potential targets for the modulation of diseases caused by retroviruses. During the retroviral life cycle, numerous cellular factors interact with the virus and play an essential role in infection. Cultured embryonic stem (ES) cells are susceptible to retroviral infection, therefore providing access to all of the genes required for this process to take place. In order to identify the host factors involved in retroviral infection, we designed and implemented a scheme for identifying ES cells that are resistant to retroviral infection and subsequent cloning of the mutated gene. RESULTS: A library of mutant ES cells was established by genome wide insertional mutagenesis in Blm-deficient ES cells, and a screen was performed by superinfection of the library at high multiplicity with a recombinant retrovirus carrying a positive and negative selection cassette. Stringent negative selection was then used to exclude the infected ES cells. We successfully recovered five independent clones of ES cells that are resistant to retroviral infection. Analysis of the mutations in these clones revealed four different homozygous and one compound heterozygous mutation in the mCat-1 locus, which confirms that mCat-1 is the ecotropic murine leukemia virus receptor in ES cells. CONCLUSION: We have demonstrated the feasibility and reliability of this recessive genetic approach to identifying critical genes required for retroviral infection in ES cells; the approach provides a unique opportunity to recover other cellular factors required for retroviral infection. The resulting insertionally mutated Blm-deficient ES cell library might also provide access to essential host cell components that are required for infection and replication for other types of virus. PMID- 17407600 TI - Enhanced NF kappa B and AP-1 transcriptional activity associated with antiestrogen resistant breast cancer. AB - BACKGROUND: Signaling pathways that converge on two different transcription factor complexes, NFkappaB and AP-1, have been identified in estrogen receptor (ER)-positive breast cancers resistant to the antiestrogen, tamoxifen. METHODS: Two cell line models of tamoxifen-resistant ER-positive breast cancer, MCF7/HER2 and BT474, showing increased AP-1 and NFkappaB DNA-binding and transcriptional activities, were studied to compare tamoxifen effects on NFkappaB and AP-1 regulated reporter genes relative to tamoxifen-sensitive MCF7 cells. The model cell lines were treated with the IKK inhibitor parthenolide (PA) or the proteasome inhibitor bortezomib (PS341), alone and in combination with tamoxifen. Expression microarray data available from 54 UCSF node-negative ER-positive breast cancer cases with known clinical outcome were used to search for potential genes signifying upregulated NFkappaB and AP-1 transcriptional activity in association with tamoxifen resistance. The association of these genes with patient outcome was further evaluated using node-negative ER-positive breast cancer cases identified from three other published data sets (Rotterdam, n = 209; Amsterdam, n = 68; Basel, n = 108), each having different patient age and adjuvant tamoxifen treatment characteristics. RESULTS: Doses of parthenolide and bortezomib capable of sensitizing the two endocrine resistant breast cancer models to tamoxifen were capable of suppressing NFkappaB and AP-1 regulated gene expression in combination with tamoxifen and also increased ER recruitment of the transcriptional co-repressor, NCoR. Transcript profiles from the UCSF breast cancer cases revealed three NFkappaB and AP-1 upregulated genes--cyclin D1, uPA and VEGF--capable of dichotomizing node-negative ER-positive cases into early and late relapsing subsets despite adjuvant tamoxfien therapy and most prognostic for younger age cases. Across the four independent sets of node-negative ER-positive breast cancer cases (UCSF, Rotterdam, Amsterdam, Basel), high expression of all three NFkappaB and AP-1 upregulated genes was associated with earliest metastatic relapse. CONCLUSION: Altogether, these findings implicate increased NFkappaB and AP-1 transcriptional responses with tamoxifen resistant breast cancer and early metastatic relapse, especially in younger patients. These findings also suggest that agents capable of preventing NFkappaB and AP-1 gene activation may prove useful in restoring the endocrine responsiveness of such high-risk ER-positive breast cancers. PMID- 17407601 TI - Dependence of paracentric inversion rate on tract length. AB - BACKGROUND: We develop a Bayesian method based on MCMC for estimating the relative rates of pericentric and paracentric inversions from marker data from two species. The method also allows estimation of the distribution of inversion tract lengths. RESULTS: We apply the method to data from Drosophila melanogaster and D. yakuba. We find that pericentric inversions occur at a much lower rate compared to paracentric inversions. The average paracentric inversion tract length is approx. 4.8 Mb with small inversions being more frequent than large inversions. If the two breakpoints defining a paracentric inversion tract are uniformly and independently distributed over chromosome arms there will be more short tract-length inversions than long; we find an even greater preponderance of short tract lengths than this would predict. Thus there appears to be a correlation between the positions of breakpoints which favors shorter tract lengths. CONCLUSION: The method developed in this paper provides the first statistical estimator for estimating the distribution of inversion tract lengths from marker data. Application of this method for a number of data sets may help elucidate the relationship between the length of an inversion and the chance that it will get accepted. PMID- 17407602 TI - Evolution and medicine: the long reach of "Dr. Darwin". AB - In this review we consider the new science of Darwinian medicine. While it has often been said that evolutionary theory is the glue that holds the disparate branches of biological inquiry together and gives them direction and purpose, the links to biomedical inquiry have only recently been articulated in a coherent manner. Our aim in this review is to make clear first of all, how evolutionary theory is relevant to medicine; and secondly, how the biomedical sciences have enriched our understanding of evolutionary processes. We will conclude our review with some observations of the philosophical significance of this interplay between evolutionary theory and the biomedical sciences. PMID- 17407603 TI - Human sterile alpha motif domain 9, a novel gene identified as down-regulated in aggressive fibromatosis, is absent in the mouse. AB - BACKGROUND: Neoplasia can be driven by mutations resulting in dysregulation of transcription. In the mesenchymal neoplasm, aggressive fibromatosis, subtractive hybridization identified sterile alpha motif domain 9 (SAMD9) as a substantially down regulated gene in neoplasia. SAMD9 was recently found to be mutated in normophosphatemic familial tumoral calcinosis. In this study, we studied the gene structure and function of SAMD9, and its paralogous gene, SAMD9L, and examined these in a variety of species. RESULTS: SAMD9 is located on human chromosome 7q21.2 with a paralogous gene sterile alpha motif domain 9 like (SAMD9L) in the head-to-tail orientation. Although both genes are present in a variety of species, the orthologue for SAMD9 is lost in the mouse lineage due to a unique genomic rearrangement. Both SAMD9 and SAMD9L are ubiquitously expressed in human tissues. SAMD9 is expressed at a lower level in a variety of neoplasms associated with beta-catenin stabilization, such as aggressive fibromatosis, breast, and colon cancers. SAMD9 and SAMD9L contain an amino-terminal SAM domain, but the remainder of the predicted protein structure does not exhibit substantial homology to other known protein motifs. The putative protein product of SAMD9 localizes to the cytoplasm. In vitro data shows that SAMD9 negatively regulates cell proliferation. Over expression of SAMD9 in the colon cancer cell line, SW480, reduces the volume of tumors formed when transplanted into immune deficient mice. CONCLUSION: SAMD9 and SAMD9L are a novel family of genes, which play a role regulating cell proliferation and suppressing the neoplastic phenotype. This is the first report as far as we know about a human gene that exists in rat, but is lost in mouse, due to a mouse specific rearrangement, resulting in the loss of the SAMD9 gene. PMID- 17407604 TI - Imprint cytology on microcalcifications excised by vacuum-assisted breast biopsy: a rapid preliminary diagnosis. AB - BACKGROUND: To evaluate imprint cytology in the context of specimens with microcalcifications derived from Vacuum-Assisted Breast Biopsy (VABB). PATIENTS AND METHODS: A total of 93 women with microcalcifications BI-RADS 3 and 4 underwent VABB and imprint samples were examined. VABB was performed on Fischer's table using 11-gauge Mammotome vacuum probes. A mammogram of the cores after the procedure confirmed the excision of microcalcifications. For the application of imprint cytology, the cores with microcalcifications confirmed by mammogram were gently rolled against glass microscope slides and thus imprint smears were made. For rapid preliminary diagnosis Diff-Quick stain, modified Papanicolaou stain and May Grunwald Giemsa were used. Afterwards, the core was dipped into a CytoRich Red Collection fluid for a few seconds in order to obtain samples with the use of the specimen wash. After the completion of cytological procedures, the core was prepared for routine histological study. The pathologist was blind to the preliminary cytological results. The cytological and pathological diagnoses were comparatively evaluated. RESULTS: According to the pathological examination, 73 lesions were benign, 15 lesions were carcinomas (12 ductal carcinomas in situ, 3 invasive ductal carcinomas), and 5 lesions were precursor: 3 cases of atypical ductal hyperplasia (ADH) and 2 cases of lobular neoplasia (LN). The observed sensitivity and specificity of the cytological imprints for cancer were 100% (one sided, 97.5% CI: 78.2%-100%). Only one case of ADH could be detected by imprint cytology. Neither of the two LN cases was detected by the imprints. The imprints were uninformative in 11 out of 93 cases (11.8%). There was no uninformative case among women with malignancy. CONCLUSION: Imprint cytology provides a rapid, accurate preliminary diagnosis in a few minutes. This method might contribute to the diagnosis of early breast cancer and possibly attenuates patients' anxiety. PMID- 17407605 TI - Doctor and practice characteristics associated with differences in patient evaluations of general practice. AB - BACKGROUND: Variation in patients' evaluation due to general practitioner (GP) and practice factors may provide information useful in a quality improvement context. However, the extent to which differences in patients' evaluation of the GPs are associated with differences in GP and practice characteristics must also be ascertained in order to facilitate comparison of adjusted patient evaluations between GPs. The aim of this study was to determine such associations in a setting where GPs serve a list of patients and act as gatekeepers. METHODS: We carried out a patient evaluation survey among voluntarily participating GPs using the EUROPEP questionnaire, which produced 28,260 patient evaluations (response rate 77.3%) of 365 GPs. In our analyses we compared the prevalence of positive evaluations in groups of GPs. RESULTS: Our principal finding was a negative association between the GP's age and the evaluation of all aspects, except accessibility. We also found an association between the way the practice was organised and the patients' evaluation of accessibility, with GPs in single handed practices getting far the most positive evaluations. Long weekly working hours were associated with more positive evaluations of all dimensions except accessibility, whereas more than 0.5 full-time employees per GP, a higher number of listed patients per GP and working in a training practice were associated with negative evaluation of accessibility. CONCLUSION: GP characteristics are mainly associated with patients' experience of interpersonal aspects of care, while practice characteristics are associated with evaluation of accessibility. These differences need to be accounted for when comparing patient evaluations of different practices. PMID- 17407606 TI - ASH structure alignment package: sensitivity and selectivity in domain classification. AB - BACKGROUND: Structure alignment methods offer the possibility of measuring distant evolutionary relationships between proteins that are not visible by sequence-based analysis. However, the question of how structural differences and similarities ought to be quantified in this regard remains open. In this study we construct a training set of sequence-unique CATH and SCOP domains, from which we develop a scoring function that can reliably identify domains with the same CATH topology and SCOP fold classification. The score is implemented in the ASH structure alignment package, for which the source code and a web service are freely available from the PDBj website http://www.pdbj.org/ASH/. RESULTS: The new ASH score shows increased selectivity and sensitivity compared with values reported for several popular programs using the same test set of 4,298,905 structure pairs, yielding an area of .96 under the receiver operating characteristic (ROC) curve. In addition, weak sequence homologies between similar domains are revealed that could not be detected by BLAST sequence alignment. Also, a subset of domain pairs is identified that exhibit high similarity, even though their CATH and SCOP classification differs. Finally, we show that the ranking of alignment programs based solely on geometric measures depends on the choice of the quality measure. CONCLUSION: ASH shows high selectivity and sensitivity with regard to domain classification, an important step in defining distantly related protein sequence families. Moreover, the CPU cost per alignment is competitive with the fastest programs, making ASH a practical option for large scale structure classification studies. PMID- 17407607 TI - Thoracic myelopathy caused by ossification of ligamentum flavum of which fluorosis as an etiology factor. PMID- 17407608 TI - Process and formulation variables in the preparation of injectable and biodegradable magnetic microspheres. AB - The aim of this study was to prepare biodegradable sustained release magnetite microspheres sized between 1 to 2 microm. The microspheres with or without magnetic materials were prepared by a W/O/W double emulsion solvent evaporation technique using poly(lactide-co-glycolide) (PLGA) as the biodegradable matrix forming polymer. Effects of manufacturing and formulation variables on particle size were investigated with non-magnetic microspheres. Microsphere size could be controlled by modification of homogenization speed, PLGA concentration in the oil phase, oil phase volume, solvent composition, and polyvinyl alcohol (PVA) concentration in the outer water phase. Most influential were the agitation velocity and all parameters that influence the kinematic viscosity of oil and outer water phase, specifically the type and concentration of the oil phase. The magnetic component yielding homogeneous magnetic microspheres consisted of magnetite nanoparticles of 8 nm diameter stabilized with a polyethylene glycole/polyacrylic acid (PEG/PAA) coating and a saturation magnetization of 47.8 emu/g. Non-magnetic and magnetic microspheres had very similar size, morphology, and size distribution, as shown by scanning electron microscopy. The optimized conditions yielded microspheres with 13.7 weight% of magnetite and an average diameter of 1.37 microm. Such biodegradable magnetic microspheres seem appropriate for vascular administration followed by magnetic drug targeting. PMID- 17407610 TI - Exacerbations of chronic obstructive pulmonary disease: when are antibiotics indicated? A systematic review. AB - BACKGROUND: For decades, there is an unresolved debate about adequate prescription of antibiotics for patients suffering from exacerbations of chronic obstructive pulmonary disease (COPD). The aim of this systematic review was to analyse randomised controlled trials investigating the clinical benefit of antibiotics for COPD exacerbations. METHODS: We conducted a systematic review of randomised, placebo-controlled trials assessing the effects of antibiotics on clinically relevant outcomes in patients with an exacerbation. We searched bibliographic databases, scrutinized reference lists and conference proceedings and asked the pharmaceutical industry for unpublished data. We used fixed-effects models to pool results. The primary outcome was treatment failure of COPD exacerbation treatment. RESULTS: We included 13 trials (1557 patients) of moderate to good quality. For the effects of antibiotics on treatment failure there was much heterogeneity across all trials (I(2) = 75%) [corrected] Meta regression revealed severity of exacerbation as significant explanation for this heterogeneity (p = 0.038) [corrected] Antibiotics did not reduce treatment failures in outpatients with mild to moderate exacerbations (pooled odds ratio 1.81, 95% CI 0.55-1.18, I(2) = 13%) [corrected] Inpatients with severe exacerbations had a substantial benefit on treatment failure rates (pooled odds ratio of 0.25, 95% CI 0.16-0.39, I2 = 0%; number-needed to treat of 4, 95% CI 3 5) and on mortality (pooled odds ratio of 0.20, 95% CI 0.06-0.62, I2 = 0%; number needed to treat of 14, 95% CI 12-30). CONCLUSION: Antibiotics effectively reduce treatment failure and mortality rates in COPD patients with severe exacerbations. For patients with mild to moderate exacerbations, antibiotics may not be generally indicated and further research is needed to guide antibiotic prescription in these patients. PMID- 17407609 TI - Identification and characterization of insect-specific proteins by genome data analysis. AB - BACKGROUND: Insects constitute the vast majority of known species with their importance including biodiversity, agricultural, and human health concerns. It is likely that the successful adaptation of the Insecta clade depends on specific components in its proteome that give rise to specialized features. However, proteome determination is an intensive undertaking. Here we present results from a computational method that uses genome analysis to characterize insect and eukaryote proteomes as an approximation complementary to experimental approaches. RESULTS: Homologs in common to Drosophila melanogaster, Anopheles gambiae, Bombyx mori, Tribolium castaneum, and Apis mellifera were compared to the complete genomes of three non-insect eukaryotes (opisthokonts) Homo sapiens, Caenorhabditis elegans and Saccharomyces cerevisiae. This operation yielded 154 groups of orthologous proteins in Drosophila to be insect-specific homologs; 466 groups were determined to be common to eukaryotes (represented by three opisthokonts). ESTs from the hemimetabolous insect Locust migratoria were also considered in order to approximate their corresponding genes in the insect specific homologs. Stress and stimulus response proteins were found to constitute a higher fraction in the insect-specific homologs than in the homologs common to eukaryotes. CONCLUSION: The significant representation of stress response and stimulus response proteins in proteins determined to be insect-specific, along with specific cuticle and pheromone/odorant binding proteins, suggest that communication and adaptation to environments may distinguish insect evolution relative to other eukaryotes. The tendency for low Ka/Ks ratios in the insect specific protein set suggests purifying selection pressure. The generally larger number of paralogs in the insect-specific proteins may indicate adaptation to environment changes. Instances in our insect-specific protein set have been arrived at through experiments reported in the literature, supporting the accuracy of our approach. PMID- 17407611 TI - A spatio-temporal mining approach towards summarizing and analyzing protein folding trajectories. AB - Understanding the protein folding mechanism remains a grand challenge in structural biology. In the past several years, computational theories in molecular dynamics have been employed to shed light on the folding process. Coupled with high computing power and large scale storage, researchers now can computationally simulate the protein folding process in atomistic details at femtosecond temporal resolution. Such simulation often produces a large number of folding trajectories, each consisting of a series of 3D conformations of the protein under study. As a result, effectively managing and analyzing such trajectories is becoming increasingly important. In this article, we present a spatio-temporal mining approach to analyze protein folding trajectories. It exploits the simplicity of contact maps, while also integrating 3D structural information in the analysis. It characterizes the dynamic folding process by first identifying spatio-temporal association patterns in contact maps, then studying how such patterns evolve along a folding trajectory. We demonstrate that such patterns can be leveraged to summarize folding trajectories, and to facilitate the detection and ordering of important folding events along a folding path. We also show that such patterns can be used to identify a consensus partial folding pathway across multiple folding trajectories. Furthermore, we argue that such patterns can capture both local and global structural topology in a 3D protein conformation, thereby facilitating effective structural comparison amongst conformations. We apply this approach to analyze the folding trajectories of two small synthetic proteins-BBA5 and GSGS (or Beta3S). We show that this approach is promising towards addressing the above issues, namely, folding trajectory summarization, folding events detection and ordering, and consensus partial folding pathway identification across trajectories. PMID- 17407613 TI - Atopic dermatitis: insights from linkage overlap and disease co-morbidity. AB - Atopic dermatitis (AD) is a strongly heritable, chronic relapsing dermatosis that frequently co-occurs with other atopic phenotypes including asthma and allergic rhinitis (the so-called atopic triad disorders). However, despite high levels of co-morbidity, relatively low levels of genomic co-incidence have been observed between atopic triad disorders. Conversely, current mapping data have revealed a striking pattern of co-localisation between AD disease loci and those mapped using another chronic dermatological disease - psoriasis. In this review, we examine the evidence for co-localisation between AD and a range of atopic, infectious, inflammatory and autoimmune diseases, and consider the implications of these data for the AD disease concept and future research in the field. PMID- 17407612 TI - Factors associated with physiotherapy provision in a population of elderly nursing home residents; a cross sectional study. AB - BACKGROUND: Although physiotherapy (PT) plays an important role in improving activities of daily living (ADL functioning) and discharge rates, it is unclear how many nursing home residents receive treatment. Furthermore, there is a lack of insight into the determinants that influence the decision for treatment. In this study, we investigated how many nursing home residents receive PT. In addition, we analysed the factors that contribute to the variation in the provision of PT both between nursing homes and between residents. METHODS: A random sample of 600 elderly residents was taken from a random sample of 15 nursing homes. Residents had to be admitted for rehabilitation or for long-term care. Data were collected through interviews with the nursing home physician and the physiotherapist. Multilevel analysis was used to define the variation in the provision of PT and the factors that are associated with the question whether a resident receives PT or not. Furthermore the amount of PT provided was analysed and the factors that are associated with this. RESULTS: On average 69% of the residents received PT. The percentage of patients receiving treatment differed significantly across nursing homes, and especially the number of physiotherapists available, explained this difference between nursing homes. Residents admitted to a somatic ward for rehabilitation, and male residents in general, were most likely to receive PT. Residents who were treated by a physiotherapist received on average 55 minutes (sd 41) treatment a week. Residents admitted for rehabilitation received more PT a week, as were residents with a status after a total hip replacement. CONCLUSION: PT is most likely to be provided to residents on a somatic ward, recently admitted for rehabilitation to a nursing home, which has a relatively large number of physiotherapists. This suggests a potential under-use of PT for long-term residents with cognitive problems. It is recommended that physiotherapists reconsider which residents may benefit from treatment. This may require a shift in the focus of physiotherapists from 'recovery and discharge' to 'quality of life and well-being'. PMID- 17407614 TI - A longitudinal study of personality and major depression in a population-based sample of male twins. AB - BACKGROUND: The relationship between personality and psychiatric illness is complex. It is not clear whether one directly causes the other. METHOD: In a population-based sample of male twins (n=3030), we attempted to predict major depression (MD) from neuroticism (N) and extraversion (E) and vice versa, to evaluate the causal, scar, state, and prodromal hypotheses. In a longitudinal, structural equation twin model, we decomposed the covariation between N and MD into (a) genetic and environmental factors that are common to both traits, as well as specific to each one and (b) direct causal effects of N at time 1 on subsequent MD, as well as between MD and subsequent N. RESULTS: E was negatively correlated with lifetime and one-year prevalence of MD. N predicted the new onset of MD, and was predicted by both current and past MD. It did not predict the time to onset of MD. All of the covariation between N and MD was due to additive genetic and individual-specific environmental factors shared by both traits and a direct causal path between MD and N assessed later. No genetic factors were unique to either trait. CONCLUSIONS: In men, N may be a vulnerability factor for MD but does not cause it directly. However, MD may have a direct causal effect on N. The genetic overlap between N and MD in men may be greater than in women. PMID- 17407615 TI - Severe life events predict specific patterns of change in cognitive biases in major depression. AB - BACKGROUND: A long-standing debate concerns whether dysfunctional cognitive processes and content play a causal role in the etiology of depression or more simply represent correlates of the disorder. There has been insufficient appreciation in this debate of specific predictions afforded by cognitive theory in relation to major life stress and changes in cognition over time. In this paper we present a novel perspective for investigating the etiological relevance of cognitive factors in depression. We hypothesize that individuals who experienced a severe life event prior to the onset of major depression will exhibit greater changes in dysfunctional attitudes over the course of the episode than will individuals without a severe life event. METHOD: Fifty-three participants diagnosed with major depression were assessed longitudinally, approximately 1 year apart, with state-of-the-art measures of life stress and dysfunctional attitudes. RESULTS: Depressed individuals with a severe life event prior to episode onset exhibited greater changes in cognitive biases over time than did depressed individuals without a prior severe event. These results were especially pronounced for individuals who no longer met diagnostic criteria for major depression at the second assessment. CONCLUSIONS: Specific patterns of change in cognitive biases over the course of depression as a function of major life stress support the etiological relevance of cognition in major depression. PMID- 17407616 TI - Body mass index in middle life and future risk of hospital admission for psychoses or depression: findings from the Renfrew/Paisley study. AB - BACKGROUND: There is evidence that greater body mass index (BMI) protects against depression, schizophrenia and suicide. However, there is a need for prospective studies. METHOD: We examined the association of BMI with future hospital admissions for psychoses or depression/anxiety disorders in a large prospective study of 7036 men and 8327 women. Weight and height were measured at baseline (1972-76) when participants were aged 45-64. Follow-up was for a median of 29 years. RESULTS: Greater BMI and obesity were associated with a reduced risk of hospital admission for psychoses and depression/anxiety in both genders, with the magnitude of these associations being the same for males and females. With adjustment for age, sex, smoking and social class, a 1 standard deviation (s.d.) greater BMI at baseline was associated with a rate ratio of 0.91 [95% confidence interval (CI) 0.82-1.01] for psychoses and 0.87 (95% CI 0.77-0.98) for depression/anxiety. Further adjustment for baseline psychological distress and total cholesterol did not alter these associations. CONCLUSIONS: Our findings add to the growing body of evidence that suggests that greater BMI is associated with a reduced risk of major psychiatric outcomes. Long-term follow-up of participants in randomized controlled trials of interventions that effectively result in weight loss and the use of genetic variants that are functionally related to obesity as instrumental variables could help to elucidate whether these associations are causal. PMID- 17407617 TI - Prevalence of depression in older people in England and Wales: the MRC CFA Study. AB - BACKGROUND: Depression in old age is an important public health problem. The aims of this study were to report the prevalence of depression in the Medical Research Council Cognitive Function and Ageing Study (MRC CFAS), a community-based, cohort. METHOD: Following screening of 13 004 people aged 65 and over from a population base, a stratified random subsample of 2640 participants received the Geriatric Mental State (GMS) examination and were diagnosed using the Automated Geriatric Examination for Computer-Assisted Taxonomy (AGECAT) algorithm. RESULTS: The prevalence of depression was 8.7% [95% confidence interval (CI) 7.3-10.2], increasing to 9.7% if subjects with concurrent dementia were included. Depression was more common in women (10.4%) than men (6.5%) and was associated with functional disability, co-morbid medical disorder, and social deprivation. Prevalence remained high into old age, but after adjustment for other associated factors, it was lower in the older age groups. CONCLUSIONS: The prevalence of depression in the elderly is high and remains high into old age, perhaps due to increased functional disability. PMID- 17407618 TI - Work stress precipitates depression and anxiety in young, working women and men. AB - BACKGROUND: Rates of depression have been rising, as have rates of work stress. We tested the influence of work stress on diagnosed depression and anxiety in young working adults. METHOD: Participants were enrolled in the Dunedin study, a 1972-1973 longitudinal birth cohort assessed most recently in 2004-2005, at age 32 (n=972, 96% of 1015 cohort members still alive). Work stress (psychological job demands, work decision latitude, low work social support, physical work demands) was ascertained by interview. Major depressive disorder (MDD) and generalized anxiety disorder (GAD) were ascertained using the Diagnostic Interview Schedule (DIS) and diagnosed according to DSM-IV criteria. RESULTS: Participants exposed to high psychological job demands (excessive workload, extreme time pressures) had a twofold risk of MDD or GAD compared to those with low job demands. Relative risks (RRs) adjusting for all work characteristics were: 1.90 [95% confidence interval (CI) 1.22-2.98] in women, and 2.00 (95% CI 1.13-3.56) in men. Analyses ruled out the possibility that the association between work stress and disorder resulted from study members' socio-economic position, a personality tendency to report negatively, or a history of psychiatric disorder prior to labour-market entry. Prospective longitudinal analyses showed that high-demand jobs were associated with the onset of new depression and anxiety disorder in individuals without any pre-job history of diagnosis or treatment for either disorder. CONCLUSIONS: Work stress appears to precipitate diagnosable depression and anxiety in previously healthy young workers. Helping workers cope with work stress or reducing work stress levels could prevent the occurrence of clinically significant depression and anxiety. PMID- 17407620 TI - Psychological distress, physical illness and risk of myocardial infarction in the Caerphilly study. AB - BACKGROUND: Studies have found associations between psychological distress (PD) and increased risk of myocardial infarction (MI). However, it is not clear whether the relationship reflects the subtle influence of pre-existing illness on both PD and MI. This study examines the association between PD and MI in a prospective epidemiological study of 1864 middle-aged men to examine if the association is explained by existing illness. METHOD: This study was a prospective cohort study modelling the association between PD, measured using the 30-item General Health Questionnaire (GHQ) and non-fatal myocardial infarction (NFMI) and fatal/non-fatal myocardial infarction (FNFMI). The relationship was modelled in a series of logistic regression models adjusted for age, then cigarette smoking, then social position, and finally for all sociodemographic characteristics, coronary heart disease (CHD) risk factors, and baseline CHD. RESULTS: PD was associated with a 70% and 68% increased risk of NFMI and FNFMI in fully adjusted analysis. However, PD was not associated with an increased risk of NFMI and FNFMI in analyses excluding those with baseline CHD. Further, being psychologically distressed and physically ill was associated with a greater than twofold risk of NFMI and FNFMI, 2.37 (95% CI 1.33-4.20) and 2.33 (95% CI 1.32 4.12) respectively. CONCLUSIONS: This study suggests that PD is a moderator of the increased risk of MI associated with existing physical illness. PD in men who are physically ill is a marker of an underlying chronic physical illness. The prospective association of PD with MI is not independent of baseline physical illness. PMID- 17407619 TI - Serious psychiatric outcome of subjects prenatally exposed to diethylstilboestrol in the E3N cohort study. AB - BACKGROUND: Prenatal exposure to diethylstilboestrol (DES) may induce neurodevelopmental disturbances potentially mediating an increased risk of psychiatric disorders in exposed subjects. Most findings of an increased prevalence of psychiatric disorders in men and women prenatally exposed to DES are not easy to interpret because of selection biases. METHOD: Information on hormonal treatment during pregnancy and on offspring's medical outcome was collected from women participating in the Etude Epidemiologique de femmes de la Mutuelle Generale de l'Education Nationale (E3N) prospective cohort who completed consecutive postal questionnaires on a range of medical events since 1990. Information on hormonal treatment during pregnancy was collected in 1992 and on offspring's medical outcome in 2004. The psychiatric outcome of subjects prenatally exposed to DES was compared to that of their unexposed siblings. RESULTS: A total of 1352 mothers with DES treatment for at least one pregnancy provided information on 1680 exposed children and 1447 unexposed siblings. After adjustment for duration of follow-up, educational level, history of obstetric complication, prenatal exposure to progestagen drugs or other hormones and parental history of psychiatric hospitalization, no association was found between prenatal exposure to DES and occurrence of strictly defined serious psychiatric outcome (suicide or psychiatric hospitalization) [adjusted odds ratio (OR) 0.8, 95% confidence interval (CI) 0.5-1.2], or of broadly defined serious psychiatric outcome (same events plus psychiatric or psychological consultation) (adjusted OR 1.0, 95% CI 0.8-1.2). CONCLUSIONS: These findings suggest that the impact of prenatal DES exposure on foetal brain development, if any, is unlikely to increase the risk of serious psychiatric disorders. PMID- 17407621 TI - Psychometric properties of DSM assessments of illicit drug abuse and dependence: results from the National Epidemiologic Survey on Alcohol and Related Conditions (NESARC). AB - BACKGROUND: DSM-IV criteria for illicit drug abuse and dependence are largely based on criteria developed for alcohol use disorders and there is a lack of research evidence on the psychometric properties of these symptoms when applied to illicit drugs. METHOD: This study utilizes data on abuse/dependence criteria for cannabis, cocaine, stimulants, sedatives, tranquilizers, opiates, hallucinogens and inhalants from the National Epidemiological Survey on Alcohol and Related Conditions (NESARC, n=43 093). Analyses included factor analysis to explore the dimensionality of illicit drug abuse and dependence criteria, calculation of item difficulty and discrimination within an item response framework and a descriptive analysis of 'diagnostic orphans': individuals meeting criteria for 1-2 dependence symptoms but not abuse. Rates of psychiatric disorders were compared across groups. RESULTS: Results favor a uni-dimensional construct for abuse/dependence on each of the eight drug classes. Factor loadings, item difficulty and discrimination were remarkably consistent across drug categories. For each drug category, between 29% and 51% of all individuals meeting criteria for at least one symptom did not receive a formal diagnosis of either abuse or dependence and were therefore classified as 'orphans'. Mean rates of disorder in these individuals suggested that illicit drug use disorders may be more adequately described along a spectrum of severity. CONCLUSIONS: While there were remarkable similarities across categories of illicit drugs, consideration of item difficulty suggested that some alterations to DSM regarding the relevant severity of specific abuse and dependence criteria may be warranted. PMID- 17407622 TI - Hepatitis B virus transmission in The Netherlands: a population-based, hierarchical case-control study in a very low-incidence country. AB - We report the first population-based case-control study on acute hepatitis B in a very low-incidence country. A case was a Netherlands resident, notified between May 1999 and July 2000 with symptoms and serology compatible with acute hepatitis B. Population controls were randomly selected, with oversampling from men and persons aged 20-39 years. Risk factors were studied using logistical regression, distinguishing confounders and mediators through hierarchical analysis. Participants were 120 cases and 3948 controls. The risk of acute hepatitis B was increased in men who have sex with men, with reporting to have had more than two partners in the past 6 months the only significant risk. In children, adult females and heterosexual males, having parents born in a hepatitis B endemic country was a significant risk. For adult females and heterosexual males, this was largely explained by having a foreign partner. For children this was partly explained by parenteral exposures abroad. PMID- 17407623 TI - Transferability of economic evaluations: approaches and factors to consider when using results from one geographic area for another. AB - BACKGROUND: Geographic transferability of economic evaluation data from one country to another has the potential to make a more efficient use of national and international evaluation resources. However, inappropriate transferability of economic data can provide misleading results and lead to an inefficient use of scarce health care resources. OBJECTIVES: The objective of this study was to review, summarize and categorize the literature on: (i) factors affecting the geographic transferability of economic evaluation data; and (ii) approaches which have either been proposed or used for transferability. METHODS: A systematic literature review on transferability was conducted. Electronic databases, hand searching and bibliographic searching techniques were utilized. Inclusion criteria for the review included conceptual or empirical papers with mention of factors affecting, or approaches for, transferability of economic evaluation data across geographic locations. Exclusion criteria included papers published prior to 1966, non-English language papers, pure science studies and animal studies. Three databases were involved in the primary search: Ovid MEDLINE, EMBASE, and CINAHL. In addition to the primary search, the Heath Economic Evaluation Database (OHE HEED), the NHS EED database and the EconLit databases were searched. Transferability factors were classified into major and minor categories, a classification of alternative transferability approaches was developed, and the number of empirical studies was catalogued according to this classification. RESULTS: There is a substantial amount of literature on factors potentially affecting transferability. Based on these papers we identified 77 factors and subsequently developed a classification system which grouped these factors into five broad categories based on characteristics of the patient, the disease, the provider, the health care system and methodological conventions. Another 40 studies were identified which attempted to transfer economic evaluation data from one country to another and these were classified according to the sources for clinical efficacy, resource utilization and unit cost data. CONCLUSIONS: There is strong evidence indicating that transferability of economic evaluation data is a difficult and complex task. Approaches which have been used for transferability suggest that, at a minimum, there is a need for country-specific substitution of practice pattern data as well as unit cost data. A limitation of this review relates to the lack of empirical studies which prevents stronger conclusions regarding which transferability factors are most important to consider and under which circumstances. PMID- 17407626 TI - Escitalopram in obsessive-compulsive disorder: a randomized, placebo-controlled, paroxetine-referenced, fixed-dose, 24-week study. AB - OBJECTIVE: A randomized, placebo controlled fixed-dose trial was undertaken to determine the efficacy and tolerability of escitalopram in obsessive-compulsive disorder (OCD), using paroxetine as the active reference. RESEARCH DESIGN AND METHODS: A total of 466 adults with OCD from specialized clinical centres, psychiatric hospital departments, psychiatric practices, or general practice were randomized to one of four treatment groups: escitalopram 10 mg/day (n = 116), escitalopram 20 mg/day (n = 116), paroxetine 40 mg/day (n = 119), or placebo (n = 115) for 24 weeks. The primary efficacy endpoint was the mean change in the Yale Brown Obsessive-Compulsive Scale (Y-BOCS) total score from baseline to week 12. Secondary efficacy endpoints included remission (defined as Y-BOCS total score < or =10), NIMH-OCS, and CGI-S and CGI-I scores at weeks 12 and 24. Tolerability was based on the incidence of adverse events, and on changes in vital signs (blood pressure and pulse). Main outcome measures; RESULTS: Escitalopram 20 mg/day was superior to placebo on the primary and all secondary outcome endpoints, including remission. Escitalopram 10 mg/day and paroxetine 40 mg/day were also effective on the primary scale as well as some other outcome measures. In the escitalopram 20 mg/day group, the improvement in Y-BOCS total score was significantly better than in the placebo group as early as week 6. The most common AEs in the active treatment groups were nausea (19-27%), headache (17 22%), and fatigue (12-19%). More paroxetine-treated patients withdrew due to adverse events than escitalopram- or placebo-treated patients. CONCLUSION: Given that escitalopram 20 mg/day was associated with an earlier onset, higher response and remission rates, improved functioning, and better tolerability than the reference drug, escitalopram deserves to be considered as one of the first-line agents in the pharmacotherapy of OCD for longer-term treatment periods. PMID- 17407624 TI - Comparing the fixed combination brimonidine-timolol versus fixed combination dorzolamide-timolol in patients with elevated intraocular pressure. AB - PURPOSE: To evaluate the efficacy of fixed combination brimonidine-timolol (FCBT) versus fixed combination dorzolamide-timolol (FCDT) given twice daily in patients with primary open angle glaucoma (POAG) or ocular hypertension (OH). DESIGN: Prospective, multicentre, masked-observer, crossover comparison. PARTICIPANTS: Sixteen patients with POAG and 14 with OH. METHODS: The participants of the study were washed out from their previous medication and randomized to fixed FCBT or FCDT for the first 4-week treatment period. Subjects then were washed for 4 weeks and started on the opposite medication for the second 4-week period. Intraocular pressure (IOP) was measured with a Goldmann applanation tonometer at 8:00 a.m., 12:00 noon and 4:00 p.m. at each baseline and at the end of each treatment period. Unsolicited ocular adverse events were also recorded. MAIN OUTCOME MEASURES: Comparison of the IOP lowering effect of FCBT and FCDT. RESULTS: The baseline mean diurnal IOP for all 30 subjects (30 eyes) was 22.9 +/- 1.6 mmHg. Both fixed combinations significantly reduced IOP compared with baseline (p < 0.00001). The mean diurnal IOP following 4 weeks of therapy was 15.0 +/- 2.1 mmHg for FCBT and 15.4 +/- 2.1 mmHg for FCDT (p = 0.510). The mean diurnal IOP reduction was 7.8 +/- 1.9 mmHg for FCBT and 7.4 +/- 1.8 mmHg for FCDT (p = 0.430). Overall, 14 subjects complained about ocular adverse events: two only for FCBT, seven only for FCDT and five for both drugs. Although there was no significant difference between the number of subjects that reported ocular adverse events with FCBT (n = 7) and FCDT (n = 12) (p = 0.359), FCDT caused more ocular stinging upon instillation (n = 9) than FCBT (n = 1) (p = 0.027). CONCLUSION: This study suggests that FCBT and FCDT, each given twice daily, have similar efficacy in patients with POAG or OH. PMID- 17407625 TI - Comparison of bisacodyl and sodium picosulphate in the treatment of chronic constipation. AB - BACKGROUND: Chronic constipation is a widespread condition. Although laxatives are generally accepted as being effective treatments, few studies have made formal comparisons of their efficacy and safety in chronic use. OBJECTIVE: To compare the safety and efficacy of bisacodyl and sodium picosulphate in the treatment of chronic constipation over a 4-week period. METHODS: Patients with chronic constipation (N = 144), recruited from out-patient clinics, were analysed for safety and efficacy in this open-label, randomised, parallel-group study. Patients were treated daily for 4 weeks (bisacodyl, 5-10 mg daily: 70 patients; sodium picosulphate, 5-10 mg daily: 74 patients). Primary efficacy criteria consisted of the number of bowel movements and stool consistency. Secondary efficacy criteria were straining at stool and physicians' global efficacy assessment. Safety assessments included adverse event monitoring, tolerability and changes in laboratory parameters. RESULTS: Both treatments were equally effective in treating chronic constipation, providing sustained improvement in symptoms. Compared to baseline, there were significant (p < 0.001) improvements in stool frequency and consistency and in the occurrence of straining at 14 and 28 days for both treatment groups. Based on the physicians' global assessment, a significant improvement was observed in 74.6% (bisacodyl) and 79.2% (sodium picosulphate) of patients. Neither treatment had significant effects on serum electrolytes. There was a trend for better tolerability in patients receiving bisacodyl treatment based on the number of drug-related adverse events (bisacodyl: 7; sodium picosulphate: 14, two patients withdrawn). CONCLUSIONS: Bisacodyl and sodium picosulphate are equally well tolerated and effective in the treatment of chronic constipation over a 4-week period. PMID- 17407627 TI - Dual cholesterol inhibition with ezetimibe/simvastatin in pre-treated hypercholesterolaemic patients with coronary heart disease or diabetes mellitus: prospective observational cohort studies in clinical practice. AB - OBJECTIVE: Patients with primary hypercholesterolaemia and concomitant coronary heart disease (CHD) and/or diabetes mellitus (DM), who are at particularly high risk of cardiovascular events such as stroke or myocardial infarction, benefit from aggressive lipid lowering strategies. The present studies investigated the incremental efficacy and safety of dual cholesterol inhibition with ezetimibe/simvastatin in such high-risk patients pre-treated with statins but not reaching the 100 mg/dL (2.6 mmol/L) low density cholesterol (LDL-C) cholesterol threshold in the primary care setting. METHODS: Two open-label, prospective, non randomised, observational studies (study 1 with n = 19 194 patients, predominantly with CHD; study 2 with n = 19 484 patients, predominantly with DM). Patients received--almost all after statin pre-treatment--ezetimibe 10 mg plus simvastatin 10 mg (study 1: 15%, study 2: 16%), 20 mg (in 68% each), 40 mg (12%/10%) or 80 mg (1%/1%) as fixed dose combinations over 3 months (dosage at investigator's discretion). RESULTS: Mean LDL-C was reduced by 28%/27% (study 1/ study 2) compared with baseline values (on statin monotherapy). Mean total cholesterol was decreased by 22% in each study, mean triglycerides by 16/17%, and mean high density cholesterol (HDL-C) was increased by 9/10%. Adverse events were reported in 0.3% and 0.2% of patients, respectively. CONCLUSION: Dual cholesterol inhibition with ezetimibe/simvastatin was effective and well tolerated under real practice conditions in high-risk patients with CHD and/or DM. PMID- 17407628 TI - Asthma control in patients with asthma and allergic rhinitis receiving add-on montelukast therapy for 12 months: a retrospective observational study. AB - BACKGROUND: Montelukast, a potent leukotriene receptor antagonist, is approved for treatment of both asthma and allergic rhinitis (AR). No studies to date have examined whether montelukast can improve asthma control over a long period of time in patients with seasonal AR and asthma. OBJECTIVE: To evaluate asthma control and use of asthma-related medical resources by patients with inadequately controlled mild to moderate persistent asthma and seasonal AR who required addition of montelukast as part of routine care. METHODS: This multicenter, 24 month, pre-post retrospective observational study included patients receiving current inhaled corticosteroid (ICS) therapy (alone or in combination with long acting beta-agonist [LABA]), who received add-on treatment with montelukast for 12 consecutive months. The incidence of asthma attacks, defined as emergency department visit, hospitalization, or oral corticosteroid use for asthma, was compared for the year before and the year after addition of montelukast to therapy. RESULTS: For the 696 patients from Italy, Poland, and Spain who were included in the analyses, the proportion of patients experiencing an asthma attack declined from 31.5% in the year before to 10.1% (p < 0.001) the year after addition of montelukast to therapy. Proportions of patients with an asthma related emergency room visit, hospitalization, and oral corticosteroid use declined from 18.7% to 3.9%, from 5.2% to 1.4%, and from 17.5% to 5.9% (all p < 0.01), respectively. The incidence of these outcomes declined in all three countries, regardless of baseline asthma severity or asthma therapy (ICS alone or ICS + LABA). Important study limitations include the possibility of selection bias or missing medical chart data in this retrospective study design. Also noteworthy is the inclusion of only those patients who remained persistent with montelukast therapy. Therefore, the results of the study are relevant for patients who remain persistent with montelukast therapy. CONCLUSIONS: Addition of montelukast to current ICS therapy improved long-term asthma control and resulted in substantial reductions in asthma-related resource use by patients with mild or moderate persistent asthma and concomitant seasonal AR who were persistent with montelukast therapy in this retrospective observational study. PMID- 17407629 TI - Interpreting score differences in the SF-36 Vitality scale: using clinical conditions and functional outcomes to define the minimally important difference. AB - OBJECTIVE: To propose the minimally important difference (MID) for the SF-36 Vitality (VT) scale by evaluating the association of score differences with clinical conditions and functional outcomes. METHODS: Analyses were performed on data from the Medical Outcomes Study (n = 3445). The first analyses regressed VT scores (0-100 scale) on chronic conditions that cause fatigue in order to determine the impact of each condition on VT. The second set of analyses examined the relationship between baseline VT scores and other outcomes at baseline, 1 year, and 7-year follow-up. RESULTS: VT scores were significantly reduced in patients with anemia [5 points (95% CI 2-9 points)], CHF [6 (3-9) points], and COPD [6 (3-9) points]. Decreases in VT score were significantly associated with increased odds of negative outcomes, including inability to work due to health at baseline [OR (5 points) = 1.27 (95% CI 1.24-1.31), OR (10 points) = 1.62 (1.54 1.71)], job loss at 1 year [OR (5) = 1.13 (1.08-1.19), OR (10) = 1.28 (1.17 1.41)], hospitalization at 1 year [OR (5) = 1.08 (1.05-1.11), OR (10) = 1.17 (1.10-1.23)], short-term mortality [0-18 months-Hazard Ratio (HR) (5) = 1.10 1.71, HR (10) = 1.21-2.39, depending on VT level] and long-term mortality [19+ months-HR (5) = 1.05-1.31, HR (10) = 1.10-1.54]. The mortality risk increase was largest at low VT levels. CONCLUSIONS: VT decrements of 5-10 points were seen for diseases known to cause fatigue. Further, differences of 5-10 points in the VT score were associated with significant increased risk of negative outcomes. We recommend an MID of 5 points for analyses of groups with VT scores below average. For follow-up of individual patients, we recommend a 10-point difference as important. PMID- 17407630 TI - Safety and efficacy of newly formulated selegiline orally disintegrating tablets as an adjunct to levodopa in the management of 'off' episodes in patients with Parkinson's disease. AB - OBJECTIVE: Patients receiving levodopa for Parkinson's disease experience motor fluctuations and immobility ('off' episodes) between doses. This study assessed adjunctive Zelapar (selegiline orally disintegrating tablet (ODT)) for managing off episodes and for long-term safety. METHODS: This open-label extension evaluated long-term safety, efficacy, and tolerability of adjunctive selegiline ODT 2.5 mg in patients who completed either of two large phase 3 double-blind studies. The study was to end after 12 months but was amended to be open-ended. Investigators could increase levodopa doses and introduce controlled-release formulations of levodopa or dopamine agonists if warranted. Additionally, results of a small randomized trial of open-label selegiline ODT 1.25 mg in comparison to conventional selegiline was added only to the safety analysis. Efficacy variables included changes in daily off time and Patient's Global Impression of Improvement (PGI-I) and Clinical Global Impressions Severity of Disease (CGI-S) ratings. Safety assessments included adverse events and oropharyngeal findings. RESULTS: This study enrolled 254 patients: 248 from the large phase 3 studies (efficacy analysis) and an additional six from the prior open-label comparison (safety analysis) in order to evaluate a larger population for safety purposes. Mean reduction from baseline in daily off time was 9.4% (1.6 h) for patients previously given selegiline ODT, 6.0% (1.2 h) for those switched from placebo, and 8.1% (1.4 h) overall. PGI-I and CGI-S ratings indicated little or no change from baseline. Treatment-related adverse events occurred in 132 (52%) patients. No severe oral irritations were attributed to selegiline ODT or prompted discontinuation. CONCLUSIONS: Long-term selegiline ODT 2.5 mg/day was effective, safe, and well tolerated in patients with Parkinson's disease experiencing off episodes during levodopa therapy. PMID- 17407631 TI - Dihydroergotamine nasal spray for relief of refractory headache: a retrospective chart review. AB - OBJECTIVE: To assess the efficacy and tolerability of dihydroergotamine (DHE) nasal spray for headache treatment in patients refractory to triptans. RESEARCH DESIGN AND METHODS: In this single-center, retrospective study, charts were reviewed to identify patients who received their first prescription for 2.0 mg DHE nasal spray between 2000 and 2004. Patients who failed previous treatments with one or more triptan formulations were considered refractory to triptan treatment and were included in the study. Headache severity was assessed by the patient at the center using a visual analog scale (VAS) of 1 to 10 (10 being most severe) at baseline and 4 weeks after initiating DHE. The responses to DHE were evaluated and categorized as complete response (headache symptoms resolved), partial response (> or = 50% reduction in VAS), or unresponsive (< 50% reduction in VAS). Four weeks after DHE use, any adverse event (AE) that occurred during DHE use was reported by the patient at the center. MAIN OUTCOME MEASURES: The efficacy of DHE was determined by headache severity reductions. Tolerability was assessed in terms of AE frequency. RESULTS: A total of 97 patients met the study criteria; 13 patients were lost to follow-up. Thirty-three patients (34.0%) reported a complete response to DHE treatment, 13 (13.4%) experienced a partial response, and 38 (39.2%) were unresponsive. Seven of 97 patients (7.2%) reported AEs (e.g., nasal congestion, dysphoria) while using DHE. CONCLUSIONS: This retrospective chart review included patients who failed triptan therapy for treatment of headaches. We found that 47% of patients experienced partial to complete response to DHE treatment. Study limitations included the retrospective design, the small sample size, and the use of patient recollection to evaluate the efficacy and tolerability of DHE. Randomized, double-blind, controlled trials are warranted. PMID- 17407632 TI - In-hospital treatment of hyperglycemia: effects of intensified subcutaneous insulin treatment. AB - BACKGROUND: Hyperglycemia is common in hospitalized patients; however, glycemic control obtained during hospitalization is often suboptimal. No methods for achievement of proper glycemic control in this population have been validated in the in-hospital setting. AIMS: To study the effect of a novel intensive subcutaneous insulin protocol on the quality of in-hospital glycemic control. METHODS: Included in this prospective controlled study were all diabetic patients admitted to the internal medicine departments in a tertiary medical center during a 1-year period. The study was divided into pre-intervention (n = 94), intervention (n = 102) and post-intervention (n = 79) periods. During the intervention period all hospitalized diabetic patients with blood glucose > 200 mg/dL were treated with an intensive multi-injection protocol consisting of two or four times daily regular/NPH insulin injections. RESULTS: Mean glucose level throughout hospitalization was 178.7 +/- 47 mg/dL in the intervention period versus 198.8 +/- 60 mg/dL in the pre-intervention period (p < 0.05). During the intervention period, the difference between mean admission and discharge day glucose levels was 43 mg/dL in patients treated with four times daily insulin injections, in contrast to no change noted in the other treatment groups. During the post-intervention period the rate of implementation of the intensive protocol by the internal medicine teams declined to 47.5%, in contrast to a 78.4% implementation rate during the intervention period. This decline was associated with deterioration of glycemic control. CONCLUSIONS: The use of intensified insulin regimen improved the glycemic control of hospitalized diabetic patients. Successful incorporation of such intensive protocols into daily medical routines requires close involvement and continuous physician guidance by the hospital diabetes team. PMID- 17407633 TI - Efficacy and safety of ezetimibe co-administered with atorvastatin in untreated patients with primary hypercholesterolaemia and coronary heart disease. AB - OBJECTIVE: Combination of ezetimibe (EZE) with a statin represents an attractive strategy for cholesterol-lowering treatment, as it inhibits the two main sources of cholesterol: absorption from the intestine (inhibited by EZE) and endogenous biosynthesis (inhibited by statins). RESEARCH DESIGN AND METHODS: This multicentre, double-blind, placebo-controlled study randomised a total of 148 men and women with primary hypercholesterolaemia and coronary heart disease (CHD) to receive treatment for 6 weeks with either EZE 10 mg + atorvastatin 10 mg (EZE + ATV; n = 72) or placebo/atorvastatin 10 mg (ATV; n = 76). The primary efficacy variable was the mean percentage change in low-density lipoprotein cholesterol (LDL-C) from baseline to study endpoint. RESULTS: At 6 weeks, EZE + ATV provided a significantly greater adjusted mean change from baseline in LDL-C compared with ATV monotherapy (-50.5% vs. -36.5%; p < 0.0001), equating to an additional 14.1% reduction (95% CI -17.90, -10.19) in LDL-C. A significantly higher proportion of patients on EZE + ATV achieved the new Joint British Societies (JBS 2) recommended LDL-C goal of < 2 mmol/L (62% vs. 12% with ATV alone; p < 0.0001) and the JBS 2 minimum treatment standard of < 3 mmol/L (93% vs. 79% with ATV alone). Patients receiving EZE + ATV were 12 times more likely to reach LDL-C targets (odds ratio 12.1; 95% CI 5.8, 25.1; p < 0.0001) compared with patients receiving ATV monotherapy. Clinical chemistry profiles and the incidence of adverse events were similar in both groups. CONCLUSIONS: Adding EZE to ATV monotherapy represents an attractive and well-tolerated treatment option to bring patients at high risk of CHD to the aggressive LDL-C goals recommended by recent treatment guidelines. PMID- 17407634 TI - Onset of efficacy of tolterodine extended release in patients with overactive bladder. AB - OBJECTIVE: To assess the onset of efficacy of tolterodine extended release (ER) in patients with overactive bladder (OAB). RESEARCH DESIGN AND METHODS: A post hoc analysis was conducted using 3-day bladder diary data from a 12-week, multicenter, prospective, open-label study of tolterodine ER (4 mg qd) in patients (aged > or = 18 years) with urinary frequency (> or = 8 micturitions/24 h) and urgency (strong and sudden desire to urinate) with or without urgency urinary incontinence (UUI). MAIN OUTCOME MEASURES: Changes in micturition frequency, urgency, and UUI episodes/24 h were evaluated for treatment Days 5, 6, and 7. The percentages of patients who achieved normal micturition frequency (< 8/day) and 50%, 70%, 90%, and 100% reductions in urgency and UUI episodes (i.e., responders) were determined at Days 5, 6, and 7. Week 12 data are presented as a referent for the magnitude of treatment efficacy during Week 1. RESULTS: This analysis included 698 patients. On Day 5, there were significant reductions in all three diary variables (all p < 0.0001), and improvements continued on Days 6 and 7. More than half of the patients reported > or = 50% reductions in urgency or UUI episodes on Day 5. Responder rates for all three symptoms increased through Week 12. CONCLUSIONS: Patients with OAB experienced significant reductions in OAB symptoms as early as Day 5 of treatment with tolterodine ER. These data extend the findings of a previous analysis, in which all 3 days of the bladder diary were pooled, that demonstrated improvements in micturition frequency, urgency episodes, and UUI episodes in patients with OAB after 1 week of treatment with tolterodine ER. Limitations are that efficacy was not assessed before Day 5, this was a post hoc analysis, and the study was not placebo controlled. PMID- 17407635 TI - Clinical practice and recent recommendations in hypertension management- reporting a gap in a global survey of 1259 primary care physicians in 17 countries. AB - OBJECTIVE: High blood pressure (BP) is a leading risk factor for cardiovascular morbidity and mortality. Effective antihypertensive pharmacotherapy is available but recognition and proper management of hypertension and BP goal achievement is still poor. Therefore, it was hypothesized that physicians' attitude towards high BP, as well as patients' perception and knowledge, may influence actual management of hypertension. RESEARCH DESIGN AND METHODS: Telephone interviews were carried out with a random sample of 1259 primary care physicians in 17 countries worldwide from 12 December 2005 to 13 January 2006 using a central computer assisted telephone interview methodology (CATI). RESULTS: (1) Physicians believed that 62 +/- 21% of their patients had their BP controlled. (2) They were mostly in line with guideline recommended BP goals and 96% were aware of the elevated cardiovascular risk of hypertension, but 41% aimed to reduce BP to acceptable levels only. (3) Physicians indicated that in 41% of patients monotherapy controls BP and 71% would escalate to combination therapy after monotherapy failure. (4) 54% regard hypertension management as difficult. (5) Physicians estimated that between 60 and 70% of patients know their BP goal but thought that there was still room for improvement of hypertension management on the patient side. CONCLUSION: Although many effective treatment options for arterial hypertension exist, BP goal achievement worldwide is suboptimal, leaving patients at an unnecessary cardiovascular risk. An increase in patients' awareness and compliance together with an increased adherence of physicians to current guidelines should help in reducing the long term cardiovascular consequences of hypertension. PMID- 17407636 TI - A double-blind study evaluating the long-term safety of varenicline for smoking cessation. AB - OBJECTIVE: We assessed the safety of long-term varenicline administration for smoking cessation. METHODS: In this randomized, double-blind, multicenter trial, eligible adult smokers (18-75 years) who smoked an average of > or =10 cigarettes/day were randomized to either varenicline 1 mg twice daily (BID) or placebo for 52 weeks. Subjects made weekly clinic visits until week 8, and then every 4 weeks until week 52, with a follow-up visit at week 53. The target quit date was the morning of the week 1 clinic visit. Brief counseling was provided at each visit, and vital signs, adverse events (AEs), and smoking status were documented. Other laboratory measures were collected at specified visits. RESULTS: A total of 251 subjects were randomized to varenicline and 126 to placebo. Approximately half of the subjects in each arm completed the study (53.8% varenicline; 46.8% placebo). Treatment-emergent AEs were observed in 96.4% of varenicline- and 82.5% of placebo-treated subjects during the study. Common varenicline-associated AEs were nausea (40.2%), abnormal dreams (22.7%), and insomnia (19.1%). Most AEs were considered mild or moderate in intensity. AEs leading to discontinuation of varenicline treatment included nausea (7.6%), insomnia (3.2%), and abnormal dreams (2.4%). A single varenicline-related serious AE, bilateral subcapsular cataracts, was observed. At week 52, 7-day point prevalence abstinence rates were 36.7% (varenicline) and 7.9% (placebo). CONCLUSIONS: Varenicline 1 mg BID can be safely administered for up to 1 year. Varenicline was also a more effective smoking cessation aid than placebo throughout the study, supporting both its short- (12-week) and long-term (52 week) efficacy. PMID- 17407637 TI - The impact of anemia on quality of life and healthcare resource utilization in patients with HIV/AIDS receiving antiretroviral therapy. AB - OBJECTIVE: To understand the impact of anemia on health-related quality of life (HRQOL) and healthcare resource utilization (HRU) in patients with HIV/AIDS. METHODS: In 2003, adults with HIV/AIDS were recruited through 114 US STD/HIV clinics to complete self-administered questionnaires. Adults reporting anemia as a side-effect of antiretroviral therapy (ART) during the past month were considered anemic. HRU was evaluated by emergency room (ER) visits, days of hospitalization, and healthcare provider visits in the past 6 months. The SF-8 was used to measure HRQOL. Linear regression was used to evaluate independent effects of anemia on HRQOL and HRU controlling for demographics, years since HIV diagnosis, HIV viral load, CD4 + count, and ART use. RESULTS: 2044 patients were enrolled, with 498 (24%) experiencing anemia. Anemic patients had lower SF-8 summary scores (mental: 38.2 +/- 11.6 vs. 42.9 +/- 11.9, p < 0.001; physical: 40.1 +/- 10.0 vs. 45.5 +/- 10.4, p < 0.001) than nonanemic patients. HRU in the anemic group was significantly higher (p < 0.05), with respect to percentage of patients visiting ER, mean number of ER visits, mean number of days hospitalized, and mean number of total visits to providers. In linear regression models, anemic patients had mental and physical HRQOL scores more than 4 points lower than nonanemic patients (p < 0.001 for both). Additionally, anemic patients had 2.7 more visits to healthcare providers (p < 0.001). LIMITATIONS: Data were cross sectional, self-reported by patients, and did not include clinical measures of anemia. CONCLUSION: Based on this survey, self-reported anemia appears to be associated with worsened HRQOL and greater HRU among HIV/AIDS patients using ART. PMID- 17407638 TI - Relationship of nasal congestion with sleep, mood, and productivity. AB - OBJECTIVE: To explore the impact of nasal congestion alone relative to a full set of allergic rhinitis (AR) symptoms on sleep, fatigue, daytime somnolence, and work and school productivity in a 15-day prospective, naturalistic study. RESEARCH DESIGN AND METHODS: Patients (N = 404) received a clinical exam to confirm congestion and assess its possible causes, including confirmed allergic rhinitis. They completed a battery of patient-reported outcomes (PROs) that assess the impact of nasal congestion and morning AR symptoms on patients' reports of sleep, daytime sleepiness, fatigue, and work, school, and activity impairment. Data were analyzed using multiple regression. Each PRO was regressed separately on congestion and morning AR symptoms, controlling for patient demographics. RESULTS: Nasal congestion has a significant (p < 0.05), negative impact on patients' lives. Nasal congestion alone had only a slightly smaller negative impact on sleep adequacy relative to AR symptoms more broadly (congestion: beta = 0.137-0.534; AR: beta = 0.123-0.642). Congestion increases the likelihood of sleep problems, fatigue, shortness of breath, headache, and daytime somnolence. CONCLUSIONS: A single congestion item by itself is a statistically and substantively significant predictor of patient-relevant outcomes. Although the sample was not randomly drawn from clinics or physician offices, the consistency and strength of the findings suggest the salience of this single symptom for patients' experiences. PMID- 17407639 TI - Solifenacin treatment for overactive bladder in black patients: patient-reported symptom bother and health-related quality of life outcomes. AB - OBJECTIVE: Health-related quality of life (HRQoL) data for black patients receiving overactive bladder (OAB) treatment have not been previously reported. This study presents patient-reported outcomes, measured by symptom bother and HRQoL, in black patients participating in an open-label study of solifenacin succinate. Results are presented, as are those from the full study population. METHODS: In the 12 week, VESIcare Open-Label Trial (VOLT), patients received solifenacin 5 mg or 10 mg once daily according to an individualized, flexible dosing regimen. A post-hoc analysis assessed solifenacin efficacy and safety in blacks (n = 274). Three patient derived indices served as study endpoints. The Patient Perception of Bladder Condition (PPBC) scale assessed overall symptom bother, a visual analog scale (VAS) recorded individual symptom bother, the Overactive Bladder Questionnaire (OAB-q) measured OAB related HRQoL. RESULTS: Blacks reported significant reductions in bladder related problems based on PPBC scores (p < 0.001) and improvements in all OAB-q subscales (symptom severity, coping, concern, sleep, social, and HRQoL; p < 0.001). Based on VAS ratings, significant improvements were reported for urinary urgency, urge incontinence, frequency, and nocturia (p < 0.001 for change from baseline). Although this study was not placebo-controlled and statistical comparisons were not made, results were similar in the full study population. In total, 46% of black patients experienced adverse events (mostly anticholinergic) and 7.6% discontinued treatment as a result. CONCLUSIONS: Solifenacin treatment was perceived as offering relief from symptom bother and improving HRQoL in the black cohort from VOLT. These results are similar to those in the full VOLT population. PMID- 17407640 TI - The weekly cost of nausea and vomiting of pregnancy for women calling the Toronto Motherisk Program. AB - BACKGROUND: Nausea with or without vomiting of pregnancy (NVP) is the most common medical condition in pregnancy. NVP, even with mild symptoms, is associated with costs to society, patients, and the health care system. OBJECTIVE: The main objective of this study was to estimate the total direct and indirect costs per woman-week associated with the onset of NVP in Canada from the perspective of society. METHODS: A cost of illness study was performed to estimate the cost per woman-week associated with the onset of NVP in Canada, stratified according to the severity of NVP. Data were collected from 139 pregnant women, who called the Motherisk Program at the Hospital for Sick Children in Toronto. Results are reported in 2005 Canadian dollars. RESULTS: From the perspective of society, the total cost per woman-week was $132 ($114 indirect and $18 direct costs), $355 ($271 indirect and $84 direct costs), and $653 ($494 indirect and $159 direct costs) for women with mild, moderate, and severe NVP, respectively. Costs increased with increasing severity of NVP. CONCLUSIONS: Nausea and vomiting of pregnancy imposes an economic burden, particularly with respect to productivity losses. Limitations of the study could be potential recall bias, the unavailability of household income and follow-up interviews. PMID- 17407641 TI - Efficacy of a paracetamol and caffeine combination in the treatment of the key symptoms of primary dysmenorrhoea. AB - OBJECTIVE: Primary dysmenorrhoea is characterised by pain, cramping and backache at the time of menses. Despite the high prevalence of dysmenorrhoea, few sufficiently powered, placebo-controlled studies have examined the efficacy of over the counter analgesics in this condition. Furthermore, even fewer studies have directly examined the efficacy of analgesics on specific dysmenorrhoea symptoms. Research design and main outcome measures: This was a single-dose, placebo-controlled, double blind, crossover study carried out in 320 women with moderate-to-severe dysmenorrhoea pain. At 2 h following dosing, 1 g paracetamol plus 130 mg caffeine led to significantly greater pain relief compared to 1 g paracetamol alone (p < 0.05), 130 mg caffeine alone (p < 0.01) or placebo (p < 0.01). The combination was also significantly more effective in relieving abdominal cramping and backache compared to the other treatment arms. No major treatment related adverse events were reported during this study. CONCLUSIONS: When taken at recommended doses, both paracetamol and the combination of paracetamol and caffeine are safe and effective treatments for primary dysmenorrhoea. Consistent with results from other acute pain states, caffeine acts as an analgesic adjuvant and enhances the efficacy of paracetamol. PMID- 17407642 TI - The economic implications of the racial and ethnic disparities in the use of selective serotonin reuptake inhibitors. AB - OBJECTIVES: Previous studies have examined racial and ethnic disparities in the use of selective serotonin reuptake inhibitors (SSRI). This study aims to examine the economic implications of these disparities. RESEARCH DESIGN AND METHODS: In this retrospective observational study, the study sample was adult survey respondents with a diagnosis of depression from the Medical Expenditure Panel Survey (2002-2003). SSRI use was measured as the number of times when SSRIs were obtained. The racial and ethnic disparities in SSRI use were examined employing a negative binomial model. The economic implications of disparities were explored using a linear regression with SSRI use as an independent variable. Interaction terms between the variable for SSRI use and dummy variables for racial and ethnic groups were included to explore whether the relationships between SSRI use and health expenditures differ across racial and ethnic groups. RESULTS: The mean number of times of SSRI use was higher for non-Hispanic whites than non-Hispanic blacks (3.02 vs. 1.79; p < 0.05) and Hispanic whites (3.02 vs. 1.68; p < 0.05). These differences were still significant after adjusting for covariates (p < 0.05). In the multivariate analysis, each time of SSRI use was associated with health expenditures of $301 higher. Neither dummy variables for racial and ethnic groups nor the interaction terms between these dummy variables and the variable for SSRI use were significant. CONCLUSIONS: The lower use of SSRIs among minorities compared to non-Hispanic whites is associated with lower health expenditures among minorities. SSRI may be a proxy for improved access to health care due to under-treatment of depression in general. The main limitation of this study is that its observational nature does not allow the researchers to determine whether the association between SSRI use and the increase in health expenditures is a causal effect. PMID- 17407643 TI - A retrospective comparison of clinical outcomes and satisfaction using reconstituted recombinant gonadotropins (rFSH) or cartridge rFSH with a pen device in donor oocyte cycles. AB - BACKGROUND: Use of recombinant follicle stimulating hormone (rFSH) in a cartridge pen device offers obvious benefits for donor oocyte cycles including the administration of fewer and more patient-friendly injectable medications. METHODS: In a University-based IVF program, a total of 98 oocyte donor cycles using rFSH either reconstituted or as a pen device given to 118 recipients (eight split cycles) were retrospectively reviewed. Following discontinuation of oral contraceptive, controlled ovarian hyperstimulation was begun using either reconstituted rFSH (n = 19) or rFSH with a cartridge pen device (n = 79) (150-300 IU qd). GnRH-antagonists (Ganirelix, Organon) and an additional 75 IU of rFSH/day were begun when lead follicles were 13-14 mm in greatest diameter. The primary endpoints analyzed included cycle stimulation characteristics for each donor group and donor medication tolerance assessment with respect to each rFSH formulation, while secondary outcome measures included clinical pregnancy and implantation rates. RESULTS: Oocyte donors using the pen required significantly less rFSH (2734 IU vs. 3276 IU, p < 0.05) and scored significantly higher medication tolerance scores (3.9 +/- 0.4 vs. 3.1 +/- 0.6, p < 0.05). No other differences in cycle stimulation for oocyte donors and clinical outcomes for recipients were seen including pregnancy rates (pen, 77% vs. reconstituted, 55%, p - NS) and clinical pregnancy rates (61% vs. 45%, p - NS). However, significantly greater implantation rates (57% vs. 31%, p < 0.01) occurred in the pen group. CONCLUSION: Compared to reconstituted formulations, the pen device results in lower gonadotropin requirements and provides a simplified dosing method with better tolerance. PMID- 17407644 TI - Modelling the cost-utility of bio-electric stimulation therapy compared to standard care in the treatment of elderly patients with chronic non-healing wounds in the UK. AB - OBJECTIVE: To estimate the cost-utility of bio-electric stimulation therapy (Posifect) compared to standard care in elderly patients with chronic, non healing wounds of > 6 months duration, from the perspective of the National Health Service (NHS) in the UK. METHODS: Clinical and resource use data from a 16 week clinical evaluation of bio-electric stimulation therapy among patients who had recalcitrant wounds were combined with utility data obtained from a standard gamble analysis to construct a 16 week Markov model. The model considers the decision by a clinician to continue with a patient's previous care plan or treat with bio-electric stimulation therapy. Unit resource costs at 2005/2006 prices were applied to the resource utilisation estimates within the model, enabling the cost-utility of bio-electric stimulation therapy compared to standard care to be estimated. The acquisition cost of Posifect had not been decided at the time of performing this study. Hence, the base case analysis used a cost of 50 pounds per dressing. RESULTS: 33% of all wounds are expected to heal within 16 weeks after the start of bio-electric stimulation therapy. Consequently, using bio-electric stimulation therapy is expected to lead to a 51% decrease in the number of domiciliary clinician visits, from 4.7 to 2.3 per week. The model also showed that using bio-electric stimulation therapy instead of patients' standard care is expected to reduce the NHS cost of managing them by 16% from 2287 pounds (95% CI: 1838 pounds; 2735 pounds) to 1921 pounds (95% CI: 1609 pounds; 2233 pounds) and result in a health gain of 0.023 QALYs over 16 weeks. Hence, bio-electric stimulation therapy was found to be a dominant treatment. Sensitivity analyses demonstrated that the cost-utility of using bio-electric stimulation therapy relative to standard care is very sensitive to the acquisition cost of the therapy, the acquisition cost of patients' drugs and the number of clinician visits and less sensitive to utility values and the acquisition cost of other dressings. CONCLUSION: Within the limitations of the model, bio-electric stimulation therapy is expected to afford the NHS a cost-effective dressing compared to standard care in the management of chronic non-healing wounds of > 6 months duration. Bio-electric stimulation therapy's acquisition cost is expected to be offset by a reduction in the requirement for domiciliary clinician visits, leading to a release of NHS resources for use elsewhere in the system, thereby generating an increase in NHS efficiency. PMID- 17407645 TI - Designs of RADIANCE 1 and 2: carotid ultrasound studies comparing the effects of torcetrapib/atorvastatin with atorvastatin alone on atherosclerosis. AB - OBJECTIVE: The RADIANCE studies were designed to assess the effects of torcetrapib/atorvastatin (T/A) compared with atorvastatin alone on slowing atherosclerotic progression in patients with heterozygous familial hypercholesterolemia (RADIANCE 1) or mixed hyperlipidemia (RADIANCE 2), as measured by change in carotid intima-media thickness (CIMT). RESEARCH DESIGN AND METHODS: RADIANCE 1 and 2 were randomized, double-blind, controlled trials with a duration of 2 years. In both studies, eligible subjects began treatment with atorvastatin during a run-in period and were titrated to target LDL-C levels defined by NCEP ATP III guidelines. Subjects then proceeded to a double-blind randomized treatment period where they received one of two regimens: (i) fixed combination T/A (torcetrapib dose, 60 mg), or (ii) atorvastatin alone. In both regimens, the dose of atorvastatin was established during the run-in period (20 80 mg, RADIANCE 1; 10-80 mg RADIANCE 2). B-mode ultrasonography was performed in duplicate at baseline and at end of study, and every 6 months in between. MAIN OUTCOME MEASURES: The primary efficacy measure in both studies was the annualized rate of change in maximum CIMT of 12 pre-defined carotid segments. Further outcome measures included lipid and safety assessments. CURRENT STATUS: The number of subjects randomized was 904 in RADIANCE 1 and 752 in RADIANCE 2. Results are anticipated in 2007. PMID- 17407646 TI - An economic assessment of analogue basal-bolus insulin versus human basal-bolus insulin in subjects with type 1 diabetes in the UK. AB - BACKGROUND: A recent study demonstrated that treatment of type 1 diabetes with an analogue basal-bolus insulin regimen was associated with improved glycaemic control (HbA(1c) -0.22% points, p < 0.001), reduced risk of hypoglycaemic events (-21%, p = 0.036) and reduction in body mass index (-0.30 kg/m(2), p < 0.001) compared to a human basal-bolus regimen after 18 weeks. METHODS: A published and validated computer simulation model was used to project long-term economic and clinical outcomes in a simulated cohort of type 1 diabetes patients treated with either insulin detemir plus insulin aspart (analogue) or Neutral Protamine Hagedorn plus human soluble insulin (human), in a UK setting. Probabilities of complications and HbA(1c)-dependent adjustments were derived from major clinical and epidemiological studies. Complication and treatment costs were projected over patient lifetimes from a National Health Service perspective. Costs and clinical benefits were discounted at 3.5% annually. RESULTS: Quality-adjusted life expectancy (QALE) was 0.66 quality-adjusted life years (QALY) higher in the analogue insulin versus the human insulin group (mean +/- SD) (7.65 +/- 0.09 versus 6.99 +/- 0.08). Direct lifetime costs were 1654 pounds greater with analogue versus human insulin treatment (40,876 pounds +/- 1119 versus 39,222 pounds+/- 1141), producing an incremental cost effectiveness ratio (ICER) of 2500 pounds per QALY gained. Sensitivity analyses showed the results were robust under a range of plausible scenarios. CONCLUSIONS: Treatment with analogue insulin was associated with a decreased incidence of long-term complications and improved QALE, but slightly higher treatment costs compared to human insulin therapy. Analogue insulin treatment had an ICER within the range generally considered to represent good value for money in the UK. PMID- 17407648 TI - Progress in the treatment of type 2 diabetes: new pharmacologic approaches to improve glycemic control. AB - BACKGROUND: Type 2 diabetes mellitus (T2DM) is a leading cause of morbidity and mortality that places a substantial economic and health burden on the public. Successful management of T2DM requires strict control of glycemia as well as other risk factors to prevent disease progression. Despite the availability of multiple classes of oral antidiabetic drugs and insulin, the majority of patients fail to attain or maintain tight glycemic control over time, raising their risk of serious microvascular and macrovascular complications. SCOPE: This review briefly outlines current standards of diabetes treatment and explores several new and investigational approaches. It is based on MEDLINE literature searches (1966 August 2006) and on abstracts from the American Diabetes Association Scientific Sessions (2002-2006) and the European Association for the Study of Diabetes Annual Meetings (1998-2006). Articles concerning basic science, preclinical, and clinical trial results were selected for this review based on their originality and relevance. FINDINGS: Medical professional societies and other specialist groups have proposed a series of practical steps to enable more patients with T2DM to reach treatment goals. Among their most important recommendations is a call for new drugs to stabilize or reverse the progressive pancreatic islet-cell dysfunction that characterizes the disease. New modalities, such as incretin mimetics and DPP-4 inhibitors, are now emerging from clinical development and will provide patients with more treatment options. CONCLUSIONS: It appears likely that early and aggressive treatment with multiple drug combinations will become more common in the management of T2DM. The new treatment modalities discussed here offer hope for improved outcomes and for meeting the considerable public health challenges posed by this complex condition. However, long-term studies are needed to determine durability of treatment effects, as well as the ultimate role of these new agents in the management of patients with T2DM. PMID- 17407649 TI - Inhibition of DPP-4: a new therapeutic approach for the treatment of type 2 diabetes. AB - BACKGROUND: Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) are hormones secreted by the enteroendocrine cells of the gut in response to the ingestion of nutrients. These incretin hormones, so called because they increase insulin secretion, are key modulators of pancreatic islet hormone secretion and, thus, glucose homeostasis. The glucoregulatory effects of incretins are the basis for new therapies currently being developed for the treatment of type 2 diabetes mellitus (T2DM). Drugs that inhibit dipeptidyl peptidase-4 (DPP-4), a ubiquitous enzyme that rapidly inactivates both GLP-1 and GIP, increase active levels of these hormones and, in doing so, improve islet function and glycemic control in T2DM. SCOPE: In this review, we briefly describe (1) the role of pancreatic islet dysfunction in the onset and progression of T2DM, (2) the rationale for developing drugs that enhance incretin activity, (3) the evidence that inhibition of DPP-4 is effective in ameliorating islet dysfunction and improving glycemic control in T2DM, (4) the efficacy, safety, and tolerability of DPP-4 inhibitors as monotherapy and in combination with other antidiabetic agents, and (5) the potential utility of DPP-4 inhibitors relative to existing oral antidiabetic agents and newer antidiabetic drugs in the pipeline. The review is based upon MEDLINE literature searches (1966-August 2006) and abstracts and presentations from the American Diabetes Association Scientific Sessions (2002-2006) and the European Association for the Study of Diabetes Annual Meetings (1998-2006). Basic science, preclinical, and clinical studies and review articles published in the English language were evaluated and selected based upon consideration of their originality, relevance, and frequency of citation. FINDINGS: DPP-4 inhibitors are a new class of antidiabetogenic drugs that provide comparable efficacy to current treatments. They are effective as monotherapy in patients inadequately controlled with diet and exercise and as add on therapy in combination with metformin, thiazolidinediones, and insulin. The DPP-4 inhibitors are well tolerated, carry a low risk of producing hypoglycemia, and are weight neutral. The long-term durability of effect on glycemic control and beta-cell morphology and function remain to be established. CONCLUSIONS: Islet cell dysfunction is central to the pathogenesis of T2DM. Incretin-based therapies, including GLP 1 analogues and DPP-4 inhibitors, have been shown to restore glucose homeostasis and improve glycemic control. The DPP-4 inhibitors, which can be used as monotherapy or in combination with other antidiabetic drugs, are a promising new treatment option, especially for patients with early-stage T2DM and more severe hyperglycemia. PMID- 17407650 TI - Pancreatic islet dysfunction in type 2 diabetes: a rational target for incretin based therapies. AB - BACKGROUND: Insulin resistance alone does not result in the development of type 2 diabetes; progressive dysfunction of pancreatic islet alpha and beta cells, which results in inadequate control of hyperglycemia, must be present for the disease to develop. Because of these defects, meal-stimulated insulin secretion from beta cells is reduced and fails to meet the demands of the insulin-resistant state; in addition, glucagon production by alpha cells, which normally maintains hepatic glucose production during fasting periods, is not suppressed. This increased glucagon secretion leads to inappropriate levels of hepatic glucose output in the post-prandial state and consequently to hyperglycemia. SCOPE: This review will examine the pathophysiologic processes of type 2 diabetes and provide an overview of some of the new and emerging treatments targeting pancreatic islet dysfunction. A MEDLINE search was performed for literature published in the English language from 1966-August 2006. Abstracts and presentations from the American Diabetes Association Scientific Sessions (2002-2006) and the European Association for the Study of Diabetes Annual Meetings (1998-2006) were also searched for relevant studies. FINDINGS: Key factors in maintaining the normal balance between insulin and glucagon levels are the incretins--glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP). Intestinal absorption of glucose stimulates secretion of these hormones, which act to increase insulin and decrease glucagon secretion. Studies demonstrating that incretin activity is impaired in type 2 diabetes have led to investigations into incretin-based therapies such as incretin mimetics (analogues of GLP-1) and inhibitors of the enzyme dipeptidyl peptidase-4 (DPP-4), which inactivates native incretins. CONCLUSIONS: Pancreatic islet dysfunction is a rational target for the treatment of type 2 diabetes. Incretin mimetics and DPP-4 inhibitors have been shown to improve glucose tolerance and may also hold the potential for improving overall pancreatic islet health. It should be noted, however, that the long-term effect of these agents on glycemic control has not yet been established, and their potential impact on beta-cell function in humans remains an area of active investigation. PMID- 17407652 TI - Self-care and the English National Health Service. PMID- 17407651 TI - Oral antidiabetic agents in type 2 diabetes. AB - BACKGROUND: Oral antidiabetic agents differ with regard to mechanisms of action, hemoglobin A(1c)-lowering efficacy, safety, and tolerability. Traditional agents consist of those that enhance insulin secretion (i.e., sulfonylureas and glinides), those that enhance insulin sensitivity (i.e., metformin and the thiazolidinediones), and those that inhibit intestinal carbohydrate absorption (i.e., the alpha-glucosidase inhibitors). New oral agents include the dipeptidyl peptidase-4 (DPP-4) inhibitors, which potentiate the activity of the incretin glucagon-like peptide 1 and enhance glucose-dependent insulin secretion. SCOPE: We review the characteristics of the traditional oral agents and these newer additions to the pharmaceutical armamentarium. Abstracts and original clinical and preclinical reports in the English language were identified for review based on MEDLINE literature searches (1970-2006) and abstract collections from major diabetes meetings. CONCLUSIONS: Traditional oral agents provide significant treatment benefits for diabetic patients, including reduction in risk of microvascular complications. However, most patients with type 2 diabetes do not achieve target glycemic levels with traditional therapies, and these agents are also associated with hypoglycemia, weight gain, and poor tolerability. Oral DPP-4 inhibitors offer the potential for significant improvement in glycemic control without hypoglycemia or weight gain, although long-term durability of glycemic control (>52 weeks) has not been established. PMID- 17407653 TI - Corruption in health services. PMID- 17407654 TI - The effect of telephone reminders on attendance in respiratory outpatient clinics. AB - INTRODUCTION: Patient non-attendance is an area of concern for all health care providers. A randomized controlled trial was undertaken to investigate whether reminder telephone calls improved attendance at respiratory outpatient clinics in the English National Health Service (NHS). METHODS: Patients were randomly allocated into one of two groups, either telephone reminder group or usual care. The telephone reminder group received a reminder telephone call between 9 am and 5 pm during the week prior to their appointment. Attendance and demographic information (age, sex, diagnosis and home postcode) were recorded. RESULTS: A total of 504 patients were recruited, 258 patients were allocated to the control group and 246 patients were allocated to the telephone reminder group. Fifty eight percent of the patients allocated to the telephone reminder group were not contactable. Within the telephone reminder group, of the 104 patients who could be contacted, 86% attended. There was a significant 15% increase in attendance in the contacted group (n = 104) when compared both with the control group (71%, n = 258) and with the patients who could not be contacted (68%, n = 142) (P = 0.007; P = 0.004). It was estimated that the cost of telephoning 200 patients could be offset by preventing one non-attendance. CONCLUSION: Routine telephoning of outpatients should become standard practice if reducing non-attendance is thought to be desirable, but general practitioner (GP) referral letters and hospital records of current hospital outpatients need to include an up-to-date telephone number. Consideration should be given to 'out-of-hours' reminder calls to maximize the contact rate. PMID- 17407655 TI - How should cost-effectiveness analysis be used in health technology coverage decisions? Evidence from the National Institute for Health and Clinical Excellence approach. AB - BACKGROUND: In the National Health Service in England and Wales, technology coverage decisions are taken by the National Institute for Health and Clinical Excellence (NICE). The intention formally to apply cost-effectiveness analysis to the decision-making process distinguishes NICE from most other bodies making similar policy recommendations. We carried out a case study of the NICE Appraisals Committee to explore the influence and use of economic evaluation in the decision-making process. METHODS: Qualitative case study methodology. This involved analysis of all relevant secondary sources, observations of Appraisals Committee deliberations and interviews with a cross-section of Committee members. FINDINGS: Economic evaluation is integrated into the Committee's work. There are two main ways in which the use of economic analysis is understood by Committee members: an ordinal approach, whereby cost-effectiveness is only considered if the technology has passed a clinical effectiveness hurdle; and a framework approach, whereby the economic evaluation and model provide a structure for considering the decision problem and the evidence. These two approaches appear to operate simultaneously but are, in essence, inconsistent. CONCLUSIONS: The NICE 'experiment' has seen cost-effectiveness analysis move to the centre-ground of UK national policy deliberations regarding technology coverage. However, our case study implies that there may be room for further refinement of the appraisal process in order to resolve the observed tension between two different ways of incorporating cost-effectiveness analysis in NICE's decision-making. PMID- 17407656 TI - Prioritizing health technologies in a Primary Care Trust. AB - BACKGROUND: In the English National Health Service (NHS), Primary Care Trusts (PCTs) are responsible for commissioning health-care services on behalf of their populations. As resources are finite, decisions are required as to which services best fulfil population needs. Evidence on effectiveness varies in quality and availability. Nevertheless, decisions still have to be made. METHODS: We report the development and pilot application of a multi-criteria prioritization mechanism in an English PCT, capable of accommodating a wide variety of evidence to rank six service developments. RESULTS: The mechanism proved valuable in assisting prioritization decisions and feedback was positive. Two community-based interventions were expected to save money in the long term and were ranked at the top of the list. Based on weighted benefit score and cost, two preventive programmes were ranked third and fourth. Finally, two National Institute for Health and Clinical Excellence (NICE)-approved interventions were ranked fifth and sixth. Sensitivity analysis revealed overlap in benefit scores for some of the interventions, representing diversity of opinion among the scoring panel. CONCLUSION: The method appears to be a practical approach to prioritization for commissioners of health care, but the pilot also revealed interesting divergences in relative priority between nationally mandated service developments and local health-care priorities. PMID- 17407657 TI - Open-ended questions in surveys of patients' satisfaction with family doctors. AB - OBJECTIVE: To compare replies to open-ended and closed questions about patient satisfaction with family doctors. METHODS: Two centres of primary health care in Bialystok in northeast Poland were chosen. A self-administered questionnaire was mailed to 1000 people (500 from each centre) aged 18 years and over, randomly selected from the practices. Possible responses to the one closed question were: very good, good, bad, very bad or difficult to say. Replies to two open-ended questions were categorized as positive, neutral, negative or ambivalent. RESULTS: The response rate was 57.9%. There were some discrepancies between the closed question response and the open-ended question replies. Some of those who replied good or very good to the closed question expressed negative views in response to the two open-ended questions (14.0% and 12.4%). CONCLUSIONS: Answers to open ended questions add value to a patient satisfaction survey by providing information that answers to closed questions may not elicite. PMID- 17407658 TI - Inequality in the face of death? Public expenditure on health care for different socioeconomic groups in the last year of life. AB - OBJECTIVE: To investigate the association between public expenditure on health care in the last year of life and individual socioeconomic status in Sweden. METHODS: Population-based study of public expenditure using linked registers for all 16,617 deaths among Stockholm County Council residents in 2002 (population 1.8 million). Age-standardized, total and per capita spend were calculated by income categories, age and specialty. Multivariate analysis examined the association between socioeconomic status and public expenditure. RESULTS: County council expenditure on health care in the last year of life rose with increasing income of the deceased person. Median per capita expenditure increased from 55,417 Swedish Kronor (SEK) (US$ 7542) in the lowest income group to SEK 94,678 (US$ 12,887) in the highest. Total age-standardized spend increased by 60% across the same interval (80,227 [95% confidence interval (CI) 79,946-80,497] to SEK 127,344 [95% CI 126,969-127,719]). Expenditure decreased with increasing age over 65 years in all income groups. Higher income was independently associated with greater total public health spend in multivariate analysis, adjusting for age, sex, health-care utilization and major diagnostic groups. CONCLUSIONS: There is inequality in public expenditure on health care at the end of life across socioeconomic groups in Stockholm. This phenomenon merits attention within Sweden, and beyond, in countries with less comprehensive welfare systems. PMID- 17407659 TI - Is the metaphor of 'barriers to change' useful in understanding implementation? Evidence from general medical practice. AB - OBJECTIVES: To investigate how general medical practices in the UK react to bureaucratic initiatives, such as National Health Service (NHS) National Service Frameworks (NSFs), and to explore the value of the metaphor of 'barriers to change' for understanding this. METHODS: Interviews, non-participant observation and documentary analysis within case studies of four practices in northern England. RESULTS: The practices had not actively implemented NSFs. At interview, various 'barriers' that had prevented implementation were listed, including the complexity of the documents and lack of time. Observation suggested that these barriers were constructions used by the participants to make sense of the situation in which they found themselves. CONCLUSION: The metaphor of 'removing barriers to change' was of limited use in a context where non-implementation of policy was an emergent property of underlying organizational realities, likely to be modifiable only if these realities were addressed. PMID- 17407660 TI - How useful are Cochrane reviews in identifying research needs? AB - OBJECTIVES: To determine the extent to which reports of Cochrane reviews recommend the need for further research and, if so, the extent to which they make suggestions regarding that research. METHODS: We examined all 2535 reviews in Issue 4, 2005 of The Cochrane Library. Each review was categorized on the basis of whether a suggestion was included about specific interventions, participants, or outcome measures that should be included in future research. We also identified the frequency with which reviews conclude that no more research is needed or feasible, noted the need for further systematic reviewing, and refered to a relevant ongoing or planned study. We also report the number of studies listed in the 'Ongoing Studies' section in each review. RESULTS: Only 3.2% of reviews suggested explicitly that no more research is needed or feasible. In 82.0% of reviews, suggestions were made as to the specific interventions that need evaluating, in 30.2% the appropriate participants were suggested, and in 51.9% outcome measures were suggested. Suggestions for all three domains were made in 16.9% of the reviews. While 11.6% did not include a specific suggestion about any of these domains, 21.2% of reviews mention a relevant ongoing or planned study in one or both of the 'Implications for Research' and the 'Ongoing Studies' sections. CONCLUSIONS: Most Cochrane reviews identify residual uncertainty and are a rich source of suggestions for further health-care research. PMID- 17407661 TI - Is the British National Health Service equitable? The evidence on socioeconomic differences in utilization. AB - Is the British National Health Service (NHS) equitable? This paper considers one part of the answer to this: the utilization of the NHS by different socioeconomic groups (SEGs). It reviews recent evidence from studies on NHS utilization as a whole based on household surveys (macro-studies) and from studies of the utilization of particular services in particular areas (micro-studies). The principal conclusion from the majority of these studies is that, while the distribution of use of general practitioners (GPs) is broadly equitable, that for specialist treatment is pro-rich. Recent micro-studies of cardiac surgery, elective surgery, cancer care, preventive care and chronic care support the findings of an earlier review that use of services was higher relative to need among higher SEGs. PMID- 17407662 TI - Shifting care from hospitals to the community: a review of the evidence on quality and efficiency. AB - OBJECTIVES: A key objective in many health-care systems is to shift specialist services from acute hospitals to the community and so bring care closer to home for patients. Our aim was to review published research into the effectiveness of strategies for achieving this objective. METHODS: We conducted a 'scoping' review and qualitative data synthesis of four strategies: transfer of services from hospital to primary care; relocation of hospital services to primary care; joint working between primary and acute care; and interventions to alter the referral behaviour of primary care practitioners. RESULTS: One hundred and nineteen studies were identified and data systematically extracted. The findings suggest that transferring hospital services to primary care, and interventions that change the referral behaviour of primary care practitioners generally reduced outpatient activity but also risked reducing quality. Savings in cost were offset by increases in overall service volume and loss of economies of scale. Relocating specialists to primary care, and joint working between primary and acute care, improved access without jeopardizing quality. However, outpatient activity was rarely reduced and costs were generally increased due to loss of economies of scale. CONCLUSIONS: Our findings suggest that the policy may be effective in improving access to specialist care for patients and reducing demand on acute hospitals. There is a risk, however, that the quality of care may decline and costs may increase. PMID- 17407664 TI - Commoditization of the international teleradiology market. AB - Telemedicine improves access to medical care. However, telemedicine will also increase market volatility because of its ability to stimulate price competition and the insidious way it shifts liability for providing medical services. To cope with increased volatility, other economic sectors have evolved commodities markets by making greater use of standardized forward/future contracts. In the past, the need for medical services to be produced and consumed locally and a lack of an objective definition for medical quality, prohibited the use of forward contracts for health-care services. However, telemedicine, and the increasing use of statistical definitions of medical quality now make standardized forward contracts for health-care services conceivable. Commoditization of teleradiology would offer several advantages including increasing market transparency, a mechanism for ensuring medical quality, and a means for bringing capital into the health-care sector. To reap the benefits of a commodities market in teleradiology, the key will be for market stakeholders to overcome their fear of the unknown in order to organize a central exchange. PMID- 17407666 TI - Myth: early detection is good for everyone. PMID- 17407667 TI - What is the effectiveness of old-age mental health services? PMID- 17407669 TI - Eight Mondays: a parliamentary group ends the silence on population. PMID- 17407670 TI - Legal interpretation of the Abortion Act 1967: the role of nurses in surgical induced abortion. PMID- 17407671 TI - Development of UPCASH (Update in Contraception and Sexual Health) Flexible Continuing Medical Education. PMID- 17407672 TI - Can nurses legally perform surgical induced abortion? PMID- 17407673 TI - Migraine in pregnancy and lactation: a clinical review. PMID- 17407674 TI - Chlamydia screening and treatment in patients undergoing evacuation of retained products of conception or vaginal termination of pregnancy: an audit of the Royal College of Obstetricians and Gynaecologists' Guidelines in practice. AB - OBJECTIVE: To assess compliance with the Royal College of Obstetricians and Gynaecologists Guidelines regarding screening for and treatment of Chlamydia trachomatis before vaginal termination of pregnancy (VTOP) and surgical evacuation of retained products of conception (ERPC). METHODS: Case notes of women undergoing VTOP and ERPC in the authors' department were examined for evidence of the presence of C. trachomatis infection and identification of chlamydia risk factors, and for evidence of curative treatment for women with confirmed infection or prophylactic treatment for women at risk. RESULTS: In the VTOP group (n = 30) screening and treatment was carried out satisfactorily, aided by an assessment proforma. In the ERPC group (n = 30) there were weaknesses in the identification and treatment of patients at risk. CONCLUSIONS: We conclude that the use of a proforma helps to achieve effective screening. Adequate risk assessment of women undergoing ERPC seems to be difficult to achieve and so universal administration of prophylactic antibiotics might be a safe and efficient option until the National Chlamydia Screening Programme is established in our area. PMID- 17407676 TI - Inequalities in the provision of sexual health information for young people. AB - BACKGROUND: Sexual health has been emphasised in national and regional strategies as a target for health and social well-being. In Northern Ireland (NI), the Sexual Health Promotion Strategy concentrates on reducing the incidence of sexually transmitted infections (STIs), reducing the number of unplanned births to teenage mothers, providing appropriate, effective and equitable sexual health information, and facilitating access to sexual health services. This article reports on a study carried out within NI and explores young people's knowledge and sources of sexual health information. METHODS: School pupils aged 14-18 years (n = 414) participated in the study and a self-administered questionnaire was used to collect the data. RESULTS: Whilst approximately half of the respondents reported being sexually active, only 68.2% always used some form of contraception. In fact, 40.8% of sexually active females had used the 'morning after pill', with 37.5% of these respondents using this method more than once. The results also indicated that students receive varying amounts of sexual health information from schools resulting in inequalities with regard to sources of information. Students from a Roman Catholic religious background were more likely to receive information on sexual health from informal sources such as friends, books/magazines or television/radio than from within the school environment compared with their Protestant counterparts. CONCLUSIONS: The provision of standard and accurate information appropriate to the target population is necessary in order to reduce the increasing rates of STIs and help the Government reach their target of halving the teenage pregnancy rate by the year 2010. PMID- 17407677 TI - "Nine out of ten patients survived". PMID- 17407678 TI - Sex workers' accounts of condom use: implications for condom production, promotion and health policy. AB - OBJECTIVES: To explore sex workers' accounts of condom use and their recommendations about how condoms might be improved. METHODS: In-depth interviews were conducted with 22 female sex workers in sex work premises in London, UK and Amsterdam, The Netherlands. RESULTS: The consistent and effective use of condoms was dependent upon client selection, sex worker control of the condom, communication skills and on condom- and sex-related skills. The design of facilities, the way the encounter was structured and alarm call systems were key to generating an environment in which sex worker control of the interaction was feasible. A wide range of practices used for 'safer sex', including the use of simulated vaginal sex, skills in fitting the condom in a sexually arousing way, checking the condom placement during intercourse and holding onto the condom during withdrawal, were described but awareness of such practices was piecemeal. Several sex workers said that particular care is needed when using condoms in men with a small penis and pointed out to the authors that a smaller condom would be useful. CONCLUSIONS: The sex workers told us about the importance of environmental factors and a range of sex- and condom-related skills in 'safer sex'. Environmental 'safety' features could be addressed through a licensing system for sex work premises. Communication, condom- and sex-related skills should be more broadly disseminated through health promotion initiatives with sex workers. The issue of condom fit deserves further attention. PMID- 17407680 TI - Improving the interface between family planning services and genitourinary medicine in Portsmouth, UK: a 'virtual corridor'. AB - BACKGROUND AND METHODS: In locations where the genitourinary medicine (GUM) department and the family planning services (FPS) are on separate sites, studies have shown a low rate of attendance of clients referred between the sites. We developed a coupon system to allow clients referred from one site to be seen without a wait in the open-access clinic at the other site. Data from the first 5 months were collected. RESULTS: A total of 59 clients were referred from the FPS to GUM during the time period studied. Of these, 54 (91.5%) attended the GUM clinic. The majority (67%) were referred with symptoms suggestive of a sexually transmitted infection (STI), while other reasons for referral included contacts of STI, high-risk behaviour, and following a sexual assault. Some 40% of clients were seen within 30 minutes of their referral. Of clients referred from GUM to FPS, 12/18 (67%) attended the clinic. This is significantly lower than the attendance of clients referred to GUM (p = 0.016). For GUM to FPS referrals, 63% were seen within 30 minutes of their referral. Seven clients were referred for consideration of intrauterine device insertion for emergency contraception but only four attended. Similarly, of the four clients referred who were not using any contraception only one attended. DISCUSSION AND CONCLUSIONS: We believe the coupon system helped increase attendance rates of clients referred between the two clinics, especially by eliminating the wait in the second clinic. The lower rate of attendance in those referred for contraception is of concern. PMID- 17407681 TI - Is 3alpha-androstenol pheromone related to menstrual synchrony? AB - BACKGROUND AND METHODOLOGY: The ovarian cycles of females living and interacting together may synchronise due to pheromones released from axillary secretary glands, the highest concentration of which is produced in the mid-follicular phase, prior to ovulation. The objective of this study was to find evidence for menstrual synchrony in a group of female students living together and to obtain a correlation between the ability to smell the putative pheromone, 5alpha-androst 16-en-3alpha-ol (3alpha-androstenol), found in apocrine secretions and menstrual synchrony. This cross-sectional study involved 88 students who completed a standard questionnaire and whose sense of smell was measured using ten varying thresholds. The menstrual history, friendship scale and menstrual hygiene score was determined for the participants. RESULTS: A total of 59.1% of the subjects studied were found to have menstrual synchrony. There was no significant association between menstrual synchrony and smelling threshold. However, a significant correlation was found between menstrual synchrony and personal hygiene score (p<0.01). CONCLUSIONS: The phenomenon of menstrual synchrony may be related to various factors. The results failed to demonstrate any significant difference between synchronised and non-synchronised subjects in detecting the steroid by sense of smell. However, the odours associated with menstrual blood or vaginal discharge might have an affect on menstrual synchrony. PMID- 17407682 TI - Linking with Mtwara, Tanzania: a personal view on skill sharing for sexual health. PMID- 17407683 TI - Finding your way through Research Ethics Committees. PMID- 17407684 TI - Fat atrophy at the site of a subdermal contraceptive implant. PMID- 17407685 TI - Ectopic pregnancy with Implanon. PMID- 17407690 TI - Everything you ever wanted to know about sex toys but were too afraid to ask... PMID- 17407686 TI - Implanon failure or a natural event? PMID- 17407691 TI - The priest doctors. PMID- 17407692 TI - Pressed for time: providing the Standard Days Method and oral contraceptives in India. PMID- 17407694 TI - Increase in IUD expulsions. PMID- 17407695 TI - Oral contraceptives and cancer. PMID- 17407697 TI - Menstrual migraine. PMID- 17407698 TI - Conflicting terminology in the context of migraine. PMID- 17407700 TI - Cytology sampling using brushes. PMID- 17407701 TI - A neuromuscular syndrome caused by abnormalities in reproductive hormones. PMID- 17407702 TI - [Stem cells for treating liver diseases: the current situation and possibilities in the near future]. PMID- 17407703 TI - [Human fetal liver nonparenchymal mesenchymal stem cells differentiate into functional hepatocyte-like cells in vitro]. AB - OBJECTIVE: To induce nonparenchymal mesenchymal stem cells (NPMSCs) differentiating into functional hepatocyte-like cells in vitro, and to identify the molecular biology and functional characteristics of those hepatocyte-like cells. METHODS: Human NPMSCs were isolated and cultured with cell culture technique. NPMSCs were induced (on 1% Matrigel as a matrix and then submitted to 2.5 mmol/L AZA pretreatment for 10-12 h), by adding HGF 10 microg/L + FGF4 10microg/L + HGM into the culture medium. The characteristics of proliferation and growth of human NPMSCs were studied with methyl thiazolyl tetrazolium (MTT). The phenotypes of NPMSCs were identified by flow cytometry, immunohistochemistry and RT-PCR. Albumin (Alb) levels in culture supernatants were determined with ELISA. Staining for glycogen of undifferentiated NPMSCs and NPMSCs derivated hepatocyte-like cells was conducted with periodic acid-Schiff (PAS) test. RESULTS: Growth and division of adherent cells obtained from fetal livers were good and the amount of NPMSCs resourced from each fetus could be amplified to 109 cells after 10 serial subcultivations. The phenotype of NPMSCs was CD166 positive and CD34 negative. The shape of NPMSCs plated on Matrigel with FGF4 and HGF changed from long fusiform to polygonal or round on days 21-28. The rate of cell rounding was 40% and the ratio of dikaryocytes was 5%. Immunohistochemical and RT PCR detection showed that undifferentiated NPMSCs expressed few alpha fetoprotein (AFP) and AFP mRNA, and did not express any of the liver-specific transcription factors or cytoplasmic markers. Many cells in early induction expressed GATA4, AFP and CK18 proteins and their mRNAs, and their expressions were reduced at the late induction, but the expressions of Alb, CK18, GST-and hepatocyte transcription factor HNF1increased gradually. The ratio of Alb and CK18 positive cells was 60%. Undifferentiated NPMSCs did not produce Alb. Alb production by induced NPMSCs increased in a time-dependent manner. Glycogen storage was first seen on day 14, and maximum levels were seen after day 28. CONCLUSIONS: There are MSCs among nonparenchymal cells of fetal livers. A high ratio of hepatocyte-like cells was obtained under our induction condition. NPMSCs differentiate firstly into hepatocyte precursors, and then differentiate into mature hepatocytes and hepatocyte-like cells with positive hepatocyte markers. The induced NPMSCs have hepatocyte specific functional features. PMID- 17407704 TI - [Tracing magnetically labeled mesenchymal stem cells transplanted into rat livers by MRI]. AB - OBJECTIVE: To trace magnetically labeled MSCs transplanted into the rat livers by magnetic resonance imaging (MRI). METHODS: Feridex and DAPI labeled rat mesenchymal stem cells (MSCs) were injected via portal veins into carbon tetrachloride treated rats. MRI was performed with a clinical 1.5 T MRI machine immediately before the MSCs injection and at h 1, d 3, d 7, and d 14 after the injection, and then the signal-to-noise ratio (SNR) was measured. MRI findings were compared with the liver histopathologies after the slides were stained with fluorescence dye and Prussian blue. RESULTS: The SNR for liver was 1.10+/-0.26 at hour 1, 8.18+/-1.55 at day 3, 11.08+/-1.30 at day 7, and 14.15+/-1.02 at day 14 respectively. Within 7 days after the MSCs transplantation, the SNRs of the livers were significantly lower than those before the transplantation (P less than 0.05). Histologically, the blue fluorescent particles under the fluorescence microscopy matched in distribution with the iron particles on the Prussian blue stained slides. CONCLUSION: The magnetically labeled MSCs transplanted into livers give rise to an obvious signal decrease, and can be tracked with a 1.5 T clinical MRI machine for up to 7 days after MSCs transplantation. PMID- 17407705 TI - [Human bone marrow derived multipotent adult progenitor cells differentiating into hepatocyte-like cells after being induced by co-culturing with human hepatocyte line L02]. AB - OBJECTIVE: To investigate the possibility of marrow derived multipotent adult progenitor cells (MAPCs) differentiating into hepatocytes by co-culturing with human hepatocyte line L02, and to evaluate the potential use of MAPCs in tissue engineering either experimentally or clinically. METHODS: (1) Co-culturing without cell-to-cell contact: MAPCs and L02 hepatocytes were spread on coverslips separately (both with a cell density of 1x10(5)/ml), and then they were put in a culture dish (10 cm). The expressions of Alb, AFP, CK18, and CK19 in MAPCs were detected by immunocytochemistry at different time points. A separate culture of L02 hepatocytes served as a positive control and a separate culture of MAPCs served as a negative control. (2) Co-culturing with cell-to-cell contact: MAPCs labeled with CFSE were mixed with L02 hepatocytes (both with a cell density of 1x10(4)/ml), and then the mixed cells were seeded on specific dishes for detection by laser scanning confocal microscope (LSCM). Five days later, the cells were double-stained with SABC-Cy3. The expressions of Alb, AFP, CK18 in MAPCs were observed under LSCM. Similarly, separately cultured L02 hepatocytes served as a positive control and separately cultured MAPCs served as a negative control. RESULTS: (1) Results of co-culturing without cell-to-cell contact: On the first day, the MAPCs expressed a high level of AFP. Then AFP expression tapered daily and there was hardly any expression of AFP on day 7. The expression of Alb was very weak on day 1, but increased significantly by day 3, reached its peak on day 5, and still maintained a high level on day 7. The initial expression of CK18 appeared on day 5 and reached a higher level on day 7. The expression of CK19 was always negative. The positive control cells had a high expression of Alb and CK18, while there was a weak expression of AFP and a negative expression of CK19. The negative control cells had no expressions for the four markers. (2) Results of co-culturing with cell-to-cell contact: On day 5, there were three colors of fluorescence under LSCM: yellow cells were MAPCs differentiating into hepatocytes; green cells were undifferentiated MAPCs; red cells were L02 hepatocytes. The result showed that Alb and CK18 were expressed in many cells and AFP appeared in only a few cells. CONCLUSION: Human MAPCs can be induced to differentiate into mature hepatocyte-like cells by co-culturing with L02 hepatocytes, either with or without cell-to-cell contact, but the former way may be more effective. PMID- 17407706 TI - [The relationship between the genotype of hepatitis B virus and clinical and liver pathological features of infected patients in the Zhoushan Islands, China]. AB - OBJECTIVE: To investigate the relationship between the genotypes of hepatitis B virus and the clinical and liver pathological features of patients with chronic hepatitis in the Zhoushan Islands. METHODS: One hundred eighty HBV DNA positive chronic hepatitis patients with HBV markers were enrolled in this study. They were at least second generation Zhoushan Island residents. One hundred forty seven of them were males and 33 were females with an average age of 39.0+/-11.3. Among the 180 patients, 17 had ASC, 57 had mild CHB, 48 moderate CHB, 9 severe CHB, 6 SHB, 39 LC, and 4 had HCC. The genotypes of their serum HBV were detected by using PCR integrated with Tagman MGB probe technology, and their serum HBV markers, HBV DNA and liver functions were also examined. Out of 180 patients, 129 accepted a liver biopsy. A pathological evaluation was then performed. RESULTS: HBVs of genotype C, 135 cases (75.0%), of B, 40 cases (22.2%), and of B+C, 5 cases (2.8%) were found among these 180 patients. No genotype A or D HBV were found. The proportions of genotype C virus were 7/17, 86/114, 34/39, 6/6 in ASC, CHB, LC and SHB patients. In the hepatocellular carcinoma patients, there were 2 each of genotype B and C. Among the 99 patients with genotype C HBV, 84 cases (84.8%) showed moderate and severe inflammation histologically in their livers and among the 30 patients with B, 7 cases (23.3%) showed moderate to severe inflammation in their livers (z = 6.47, P less than 0.01). The proportion of genotype C HBV was significantly different from that of genotype B HBV in those that showed moderate and severe (S3-4) liver fibrosis. In patients infected with genotype C HBV who had moderate and severe liver pathological changes, their clinical manifestations reflected better the histological alterations of their livers. CONCLUSION: Genotypes C, B and B+C HBV were found in CHB patients in the Zhoushan Islands of China, and type C was the predominant one. The liver pathological damage level of genotype C HBV infected patients is more serious than that of genotype B. PMID- 17407707 TI - [Changes of peripheral plasmacytoid dendritic cells and lymphocyte subsets in chronic hepatitis B patients treated with consensus interferon alpha]. AB - OBJECTIVE: To study the changes of peripheral plasmacytoid dendritic cells and lymphocyte subsets in patients with chronic hepatitis B (CHB) during consensus interferon (CIFN) treatment. METHODS: Twenty-three patients with CHB were treated with CIFN for 24 weeks and followed up for another 24 weeks. Peripheral plasmacytoid dendritic cells and lymphocyte subsets were measured throughout the treatment and follow-up periods. RESULTS: After CIFN treatment, 43.5% of the patients had virological and biochemical responses. The percentage and absolute number of peripheral plasmacytoid dendritic cells decreased significantly (P less than 0.05), the number of CD3+ cells, CD4+ T cells, B cells and the ratio of CD4+/CD8+ cells decreased also (P less than 0.05), but the number of CD8+ T cells and NK cells increased (P less than 0.05). CONCLUSIONS: Consistent virological and biochemical responses can be seen in some patients with CHB virus infection after CIFN treatment, and the percentage and number of their peripheral plasmacytoid dendritic cells greatly decreased, but the number of CD8+ T cells and NK cells increased. PMID- 17407708 TI - [Genetic polymorphisms of MxA protein and eIF-2a-reg2 and their responses to interferon treatment in patients with chronic hepatitis B]. AB - OBJECTIVES: To identify the host single nucleotide polymorphisms (SNP) of myxovirus resistance A (MxA) protein and eukaryote initiation factor 2alfa regulatory region 2(eIF-2a-reg2) and to predict interferon (IFN) treatment responses in patients with chronic hepatitis B. METHODS: Two hundred sixty-two patients with chronic hepatitis B (CHB) were treated with interferon alfa (IFN ) for 12 months. Six months later the therapeutic effectiveness was evaluated. All the patients had signed a formal consent form. The patients were grouped into a sustained response (SR) group and a non-sustained response (NSR) group according to their responses to the IFNa treatment. Single nucleotide polymorphisms of the antiviral protein MxA promoter -88,-123 and protein kinase(PKR) activated eIF-2a reg2 sites were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and were compared with the responsiveness to IFN treatment of these CHB patients. RESULTS: Among the 262 patients, 212 (80.9%) were non-sustained responders to IFNa and 50 (19.1%) were sustained responders. The rate of sustained responders with GT heterozygote at MxA promoter -88 was higher than that of the GG genotype (OR: 5.3, 95% CI: 2.46-11.43, P less than 0.01) and also higher than that of the TT genotype (OR: 4.1, 95% CI: 1.86-9.09, P less than 0.01). There were no statistically significant differences in IFN therapeutic effectiveness among the patients with different genotypes at MxA promoter -123, eIF-2a-reg2 and haplotypes made by MxA promoter -88 G/T, and -123 C/A alleles (P more than 0.05). CONCLUSION: Patients with GT genotype at MxA promoter -88 responded well to IFN treatment. SNP as a potential marker could be used to predict IFN treatment responses of patients with chronic hepatitis B. PMID- 17407709 TI - [Effects of pioglitazone on the morphology and the expression of connective tissue growth factor of transforming growth factor beta-induced rat hepatic stellate cells in vitro]. AB - OBJECTIVES: To observe the effects of pioglitazone on morphological changes and connective tissue growth factor (CTGF) expression of the transforming growth factor beta (TGF b)-induced rat hepatic stellate cells (HSCs) in vitro, and to investigate the anti-fibrotic mechanism of pioglitazone. METHODS: Cultured rat HSCs were divided into a no-treatment control group, a TGF b-treated group, and a TGFb plus different dosage pioglitazone-treated group. The morphological changes of the cultured HSCs were observed. The expression of CTGF was assessed by immunohistochemistry and flow cytometry. The level of collagen type III in the culture supernatant was measured by ELISA. RESULTS: TGFb induced morphological changes, and increased the expressions of CTGF and collagen type III of the HSCs (P less than 0.05). Pioglitazone prevented the TGFb induced morphological changes of the HSCs. The expression of CTGF and the levels of collagen type III in the pioglitazone group were lower than the TGF b-treated group (P less than 0.05). This prevention effect was dose-dependent (P less than 0.05). CONCLUSION: Pioglitazone blocks the excretion of CTGF and collagen type III of cultured HSCs, preventing the development of liver fibrosis. PMID- 17407710 TI - [Construction and identification of matrix metalloproteinase inhibitor-1 siRNA eukaryotic expression vectors]. AB - OBJECTIVE: To construct TIMP-1 siRNA eukaryotic expression vectors and evaluate their effect on TIMP-1 mRNA expression in hepatic stellate cells. METHODS: The combinant lone DNA with cutting sites of BamH I and Xho I enzyme according to the sequences of 447-465, 552-540 TIMP-1 of rats and nonspecific sequence were selected and cloned to pGEM-T vector and sub-cloned to pRNAT-U6.2. They were then identified by double enzyme digestion analysis and DNA sequencing. Three plasmids were transfected into T6 separately through an oligofectamine package. TIMP-1 mRNA expression was evaluated by RT-PCR. RESULTS: Targeting sequences of TIMP-1 siRNA eukaryotic expression vectors were correct. TIMP-1 mRNA expression was significantly reduced by transfecting them into the T6. CONCLUSION: We successfully constructed two TIMP-1 siRNA eukaryotic expression vectors and the transfected cells can significantly suppress the TIMP-1 expression. PMID- 17407711 TI - [Reversal of adriamycin resistance of hepatocellular carcinoma by targeting it with recombined adenovirus carrying antisense multidrug resistance gene 1 RNA]. AB - OBJECTIVE: To investigate if an adenovirus vector carrying antisense multidrug resistance gene 1 (MDR1) could reverse multidrug resistance (MDR) of HepG2/ adriamycin (ADM) cells in tumors transplanted in athymic mice. METHODS: An adenovirus vector carrying AFP promoter and antisense MDR1 was constructed. HepG2 MDR cells (HepG2/ADM) were induced by graded resistance to ADM and were subcutaneously inoculated into athymic mice to construct the transplanted tumor. After adeno-asmdr1 was injected, the volume of the transplanted tumor and the apoptotic body in the xenograft tumor cells were observed and reverse transcriptase polymerase chain reaction was employed to investigate the expression of the mdr1-mRNA from the mouse transplanted tumor cells. RESULTS: Following injection with adeno-asmdr1, the tumor volumes in this mice group did not increase. However the tumor volume in the PBS plus ADM group did increase significantly (P less than 0.05). In the tumor xenograft cells, mdr1 mRNA in the xenografts was assessed by RT-PCR and found to be reduced at week 1, and at week 4 in the ADM+asmdr1 group, but it was stable in the ADM group. It was only 20% in the ADM+asmdr1 group compared to the ADM group at the 4th week. Evidence of apoptosis was observed in the tumor xenograft cells treated with adeno-asmdr1, but there was rarely any apoptosis in the group treated with ADM and PBS. CONCLUSION: Adenovirus carrying antisense mdr1 RNA can partially reverse the MDR of HepG2/ADM cells and inhibit tumor growth by down-regulating mdr1 mRNA resulting in tumor cell apoptosis. PMID- 17407712 TI - [A study on the preventive effect of Lycium barbarum polysaccharide on the development of alcoholic fatty liver in rats and its possible mechanisms]. AB - OBJECTIVE: To study the preventive effects of Lycium barbarum polysaccharide (LBP) on the development of alcoholic fatty liver (AFL) in rats and its possible mechanisms. METHODS: One hundred twenty-five Wistar rats were randomly divided into 4 groups: a blank control group, with distilled water intragastric infusion (GI); an alcohol group, with alcohol GI; a 5% LBP plus alcohol GI group; and a 10% LBP plus alcohol GI group. Liver pathologic changes were studied together with the activity of serum ALT, AST, GGT, the activity of liver SOD, GSH-PX and the content of liver MDA, GSH, H2O2; CYP2E1 gene and protein expressions were also detected. RESULTS: At the end of ten weeks, the activity of serum AST [(132.3+/-25.7) U/L, (127.5+/-29.1)U/L] and GGT [(1.9+/-0.5)U/L, (1.8+/-0.7)U/L] of the two LBP groups were all significantly lower than those of the alcohol group [serum AST (245.7+/-32.1) and GGT (4.4+/-0.6)]. At the end of ten weeks, the content of liver MDA [(5.1+/-0.3) nmol/mg, (5.1+/-0.4) nmol/mg] and H2O2 [(135.4+/-23.5) mmol/g, (132.6+/-31.8) mmol/g] of the two LBP groups were significantly lower than those of the alcohol group [MDA (14.5+/-3.2) nmol/mgprot) and H2O2 (328.5+/-45.6)]. The activity of SOD [(206.7+/-13.2)U/L, (203.2+/-18.8)U/L], GSH-PX [(13.5+/-1.4)U/mg/min, (13.6+/-1.5)U/mg/min] and the content of GSH [(65.1+/-11.0)mg/g, (66.6+/-11.1) mg/g] of the two LBP groups were all significantly higher than those of the alcohol group [SOD (116.5+/ 13.6)U/mg/min, GSH-PX (7.2+/-1.6)U/mg/min and the content of GSH (30.5+/ 10.7)mg/g] (P less than 0.01). At the end of five weeks, levels of CYP2E1 gene and protein expression of the two LBP groups were 0.39+/-0.04, 0.40+/-0.06 and 3.49+/-0.36, 3.29+/-0.30 respectively. At the end of ten weeks, levels of CYP2E1 gene and protein expression of the two LBP groups were 0.41+/-0.05, 0.42+/-0.08 and 3.58+/-0.30, 3.36+/-0.37 respectively. They were all significantly lower than those of the alcohol group [the gene expression (5 week: 0.74+/-0.05, 10 week: 1.02+/-0.08) and the protein expression (5 week: 5.63+/-0.44, 10 week: 7.90+/ 0.26)]. There were no typical alcoholic fatty liver pathologic changes observed in the two LBP groups. CONCLUSION: LBP can effectively prevent AFL. This may be due to its effects in inhibiting the hepatocyte CYP2E1 expression and prevention of lipid peroxidation. PMID- 17407713 TI - [The protective role of adiponectin in Con A-induced mouse liver injury]. AB - OBJECTIVE: To evaluate the role of adiponectin in regulating tumor necrosis factor alpha (TNF alpha) production and preventing fulminant autoimmunological damage of hepatocytes following concanavalin A (Con A) injection into mice. METHODS: Three days after recombinant plasmids pAA-neo-mAd were injected into the mice via the tail veins, Con A was injected into the mice. Mice transfected with empty pAA-neo vector served as controls. The serum levels of alanine aminotransferase (ALT), TNF alpha and adiponectin were detected, and histological examination of livers was carried out at different time points after the Con A injection. All results were subjected to statistical analyses. RESULTS: Histological examinations showed that the damage in livers of mice with high serum adiponectin levels was milder than that of the controls. The serum levels of ALT and TNF alpha were both lower than those of the controls (P less than 0.01, respectively). Statistical analyses showed the serum levels of ALT was negatively related to the levels of adiponectin in the sera (r=-0.5034). CONCLUSION: Adiponectin is effective in protecting hepatocytes from Con A-induced immunological injury. The mechanism of this protective effect may be caused by inhibiting the synthesis and/or release of TNF alpha. PMID- 17407714 TI - [Clinical and pathological analysis of 100 cases of drug-induced liver injury]. AB - OBJECTIVE: To study the clinical and pathological features of drug-induced liver injury (DILI). METHODS: Liver specimens were obtained through needle biopsies from 100 patients with DILI. The histological preparations of the specimens were stained with haematoxylin eosin, several histochemistry methods, and immunohistochemistry stains. The pathological changes of the livers were analyzed together with the patients's clinical data. The patients were divided into two groups, an acute DILI group (n=39) and a chronic DILI group (n=61), based on their clinical courses and histological changes in their livers. In the chronic DILI group, the clinical courses were longer than 6 months and/or fibrosis or cirrhosis occurred in their liver tissues. RESULTS: Among our cases the leading cause of DILI was Chinese herb medicine, accounting for 21% of the 100 cases; steroids induced cases were 11% of the total. 78% of the patients presented elevated serum transaminases and/or jaundice. The degree of transaminases elevation and the frequency of jaundice happening in the acute group were significantly higher than those in the chronic group (P less than 0.05). The histopathological liver changes in these DILI cases included: (1) necrosis commonly occurred in acinar zone 3, (2) abundant neutrophil and/or eosinophil infiltrations, (3) hepatocytic and/or canalicular cholestasis with little or no inflammation, (4) microvesicular steatosis mixed with macrovesicular steatosis, and (5) presentation of epitheloid cell granuloma. There were no significant differences in liver histopathology between the acute and the chronic DILI groups, except that the fibrosis and the ductular proliferation were different. CONCLUSION: DILI has become a notable liver disease in mainland China, and the use of Chinese herbal medicine must be improved, standardized and regulated more closely. PMID- 17407715 TI - [Gene therapy of hepatic stem cells in treating liver diseases]. PMID- 17407716 TI - [The advancement and expectations in using hepatocytes in bioartificial livers]. PMID- 17407717 TI - [Stem cells and liver diseases]. PMID- 17407718 TI - [A study of liver cell transformation from bone marrow hematopoietic stem cells]. PMID- 17407719 TI - [Overexpression of HCCR in hepatocellular carcinoma cells and its clinical significance]. PMID- 17407720 TI - [A clinical study of thymosin alpha1 as an auxiliary in treating spontaneous peritonitis in patients with liver cirrhosis]. PMID- 17407721 TI - [A study on the role of cyclooxygenase-2 in lipopolysaccharide mediated fulminate hepatic failure]. PMID- 17407722 TI - [A study of the association between PPARr2 gene Pro12Ala polymorphism and NAFLD]. PMID- 17407723 TI - [Changes of liver fibrosis markers in nonalcoholic fatty liver disease patients]. PMID- 17407724 TI - [A 72 week follow-up study of chronic hepatitis C patients treated with peginterferon alfa-2b and ribavirin]. PMID- 17407725 TI - [Detection of hepatitis B virus cccDNA in liver tissues of patients with hepatocellular carcinoma]. PMID- 17407726 TI - [The repopulation of human L02 hepatocytes in tolerant rats treated with retrorsine]. PMID- 17407727 TI - [Transcatheter hepatic arterial chemoembolization followed by three-dimensional conformal radiotherapy for the treatment of primary hepatic cancer with portal vein tumor thrombus]. PMID- 17407728 TI - [CD4+CD25+Treg cell and its function in liver diseases: a review]. PMID- 17407729 TI - Patient Positioning 101. Time to rethink patient access in the back of ambulances. PMID- 17407732 TI - Practice what you preach. PMID- 17407733 TI - You get what you pay for. PMID- 17407734 TI - Thanks for the high. PMID- 17407735 TI - Not just trainers. PMID- 17407737 TI - Drug abuse scandals. Thieves emphasize need for proper controls. PMID- 17407738 TI - Spring-Man. What are your jittery patients going to do next? PMID- 17407739 TI - A grim surprise. "I refuse to admit that I am more than 52, even if that makes my son illegitimate."-Lady Astor. PMID- 17407740 TI - Bug Jumping. MRSA isn't just a hospital infection anymore. PMID- 17407741 TI - The break-up. Just how bad is EMS for your marriage? PMID- 17407743 TI - The risky side of response. Increasing fatalities underscore the need for reconciled safety practices of ground & air ambulances. PMID- 17407744 TI - Improving CHF care. A new algorithm for prehospital treatment. PMID- 17407745 TI - Snakebites. Prehospital assessment & treatment of envenomations. PMID- 17407746 TI - Clown down. PMID- 17407747 TI - Virtual biomedical universities and e-learning. AB - In this special issue on virtual biomedical universities and e-learning we will make a survey on the principal existing teaching applications of ICT used in medical Schools around the world. In the following we identify five types of research and experiments in this field of medical e-learning and virtual medical universities. The topics of this special issue goes from educational computer program to create and simulate virtual patients with a wide variety of medical conditions in different clinical settings and over different time frames to using distance learning in developed and developing countries program training medical informatics of clinicians. We also present the necessity of good indexing and research tools for training resources together with workflows to manage the multiple source content of virtual campus or universities and the virtual digital video resources. A special attention is given to training new generations of clinicians in ICT tools and methods to be used in clinical settings as well as in medical schools. PMID- 17407748 TI - Balancing selection. PMID- 17407749 TI - On the use of the word 'epigenetic'. AB - Over the past few years we have seen an odd change, or extension, in the use of the word 'epigenetic' when describing matters of gene regulation in eukaryotes. Although it may generally be that it is not worth arguing over definitions, this is true only insofar as the participants in the discussion know what each other means. I believe the altered use of the term carries baggage from the standard definition that can have misleading implications. Here I wish to probe our use of language in this way, and to show how such a discussion leads to some more general considerations concerning gene regulation. PMID- 17407750 TI - The auditory cortex. PMID- 17407751 TI - Mate recognition: should fly stay or should fly go? AB - Recent studies have shown that male fruit flies use close-range olfactory cues to assess the status of potential mating partners. The presence of a volatile, male derived pheromone can suppress the default male behavioral state of courting females. PMID- 17407752 TI - Animal behavior: tolerant primates cooperate best. AB - Chimpanzees and bonobos show different cooperative tendencies when the prize is easily monopolizable food: bonobos cooperate more than their less socially tolerant relatives. PMID- 17407753 TI - Intracellular motility: myosin and tropomyosin in actin cable flow. AB - A new study has found that retrograde flow of budding yeast actin cables is facilitated by myosin II but is inhibited by a specific tropomyosin isoform (Tpm2p). Budding yeast therefore contains a minimal component system for elucidating the mechanistic details of retrograde actin flow. PMID- 17407754 TI - Animal communication: big talkers and small talk. AB - Vocal tract resonances, known as formants, are important perceptual cues for the identification of human speech and animal calls. A recent study shows that monkeys can also use formants to determine the age and size of the monkey producing a call. PMID- 17407755 TI - Microtubule organization: cell shape is destiny. AB - A simple self-assembly pathway generates cytoplasmic microtubule bundles that can locate the cell center and guide spindle assembly in fission yeast. The cylindrical cell shape automatically corrects spindle orientation errors, rendering a checkpoint unnecessary. PMID- 17407756 TI - Population biology: wild origins of a model yeast. AB - Yeast is a superb laboratory model organism, but little is known about its natural lifestyle. Recent studies of wild yeast are beginning to reveal details of Saccharomyces population structure and evolution that challenge assumptions about speciation and dispersal in microbes. PMID- 17407757 TI - Biomechanics: stable running. AB - Runners need dynamic stability to maintain their gait despite uneven terrain and other disturbances. Research on guinea fowl is providing welcome new insight into how this difficult problem is solved. PMID- 17407758 TI - Membrane trafficking: three steps to fusion. AB - Membrane fusion involves the action of members of the SNARE protein family as well as Sec1/Munc18 (SM) proteins, which have been found to interact with SNAREs in three distinct ways. Recent work has established that Munc18-1 directly stimulates fusion and possibly uses all three modes of SNARE interaction. PMID- 17407759 TI - Capillary electrophoretic determination of selenocyanate and selenium and tellurium oxyanions in bacterial cultures. AB - A simple capillary zone electrophoretic method for the determination of biospherically important oxyanions of selenium (Se) and tellurium and another Se containing anion, selenocyanate, has been developed. The method uses direct UV absorption detection. Time course experiments with time slices as short as 6 min are possible. This method's detection limits and linear range compare well with other methods involving samples containing complex biological matrices. The metalloid-containing anions examined were selenocyanate, selenite, selenate, tellurite, and tellurate. We applied this method to live cultures of two different bacteria in two different growth media in time course experiments following the changes in metalloid-containing anion concentrations. The results show that this method is a useful means of following the biological processing of these analytes in bacterial cultures. PMID- 17407760 TI - Use of metallochromic dyes and potentiometric pH-meter titration to detect binding of divalent cations to "Good's" buffers: 4-morpholinepropanesulfonic acid (Mops) does not bind Mg2+. PMID- 17407761 TI - Beta cell cytoprotection using lentiviral vector-based iNOS-specific shRNA delivery. AB - Cytokine-induced beta cell pathophysiology is characterised by the induction of iNOS expression. Inhibition of iNOS expression protects beta cells from cytokine mediated destruction. The development of vector-based shRNA strategies capable of stably suppressing iNOS expression may provide a novel platform to protect beta cells from cytokine toxicity. In this report the utility of lentiviral shRNA vectors to silence iNOS expression was evaluated with respect to insulinoma cell viability, the induction of iNOS expression and the accumulation of nitrite in a cytokine-induced beta cell toxicity model. Here, we report for the first time on the use of lentiviral vector-based shRNA delivery to efficiently suppress the IL 1beta-mediated induction of iNOS expression, the accumulation of nitrite and provide significant protection against the cytotoxic effects of IL-1beta exposure. Moreover, non-specific knockdown of endogenous beta cell nNOS did not occur. PMID- 17407762 TI - Structural requirements for furin-induced cleavage and activation of Shiga toxin. AB - Shiga toxin has a protease-sensitive site in the disulfide loop region of the A chain. Cleavage of this site by furin is essential for rapid intoxication of cells by Shiga toxin. We have here investigated whether in addition to the Arg-X X-Arg sequence, there are other structural requirements in the disulfide loop region for furin cleavage. A toxin mutant (Shiga-2D toxin) still containing the consensus motif for cleavage by furin, but lacking ten amino acids in the disulfide loop, was generated. Trypsin was able to cleave Shiga-2D toxin in vitro, demonstrating that the protease-sensitive region is intact. However, Shiga 2D toxin was not efficiently cleaved by furin either in vitro or in vivo. Furthermore, unless it was precleaved with trypsin, Shiga-2D toxin was much less toxic than wild type Shiga toxin in LoVo cells expressing functional furin. In contrast, LoVo/neo cells lacking functional furin were unable to activate both wild type Shiga toxin and Shiga-2D toxin. In conclusion, an extended loop structure is required for furin-induced cleavage of Shiga toxin. PMID- 17407763 TI - Transcriptional profiling of stress response in cultured porcine islets. AB - Cell-based diabetes therapy may be achieved through xenotransplantation of adult porcine islets, but tissue quality and immunoreactivity barriers need to be overcome. Early identification and exclusion of irreversibly stressed and dying islets may improve transplant outcomes. We used oligonucleotide microarray and quantitative RT-PCR to identify molecular markers of physiological and immunological stress in porcine islets cultured under stress conditions of elevated glucose (16.7 mM), inflammatory cytokine addition (IL-1beta, TNF-alpha, and IFN-gamma), or both, for 48 h. Hyperglycemic conditions were associated with increased thioredoxin interacting protein and metabolic process mRNAs, as observed in rodent and primate species. Cytokine treatment increased expression of JAK-STAT pathway components, oxidative stress (transglutaminase 2), and beta cell dysfunction genes. Transglutaminase 2 induction is unique to porcine islets. Biomarkers involved in hyperglycemia and islet inflammation may serve as novel targets for improving and monitoring isolated porcine islet function and viability. PMID- 17407764 TI - Crystal structure of the C-terminal three-helix bundle subdomain of C. elegans Hsp70. AB - Hsp70 chaperones are composed of two domains; the 40 kDa N-terminal nucleotide binding domain (NDB) and the 30 kDa C-terminal substrate-binding domain (SBD). Structures of the SBD from Escherichia coli homologues DnaK and HscA show it can be further divided into an 18 kDa beta-sandwich subdomain, which forms the hydrophobic binding pocket, and a 10 kDa C-terminal three-helix bundle that forms a lid over the binding pocket. Across prokaryotes and eukaryotes, the NBD and beta-sandwich subdomain are well conserved in both sequence and structure. The C terminal subdomain is, however, more evolutionary variable and the only eukaryotic structure from rat Hsc70 revealed a diverged helix-loop-helix fold. We have solved the crystal structure of the C-terminal 10 kDa subdomain from Caenorhabditis elegans Hsp70 which forms a helical-bundle similar to the prokaryotic homologues. This provides the first confirmation of the structural conservation of this subdomain in eukaryotes. Comparison with the rat structure reveals a domain-swap dimerisation mechanism; however, the C. elegans subdomain exists exclusively as a monomer in solution in agreement with the hypothesis that regions out with the C-terminal subdomain are necessary for Hsp70 self association. PMID- 17407765 TI - Microarray multiplex assay for the simultaneous detection and discrimination of hepatitis B, hepatitis C, and human immunodeficiency type-1 viruses in human blood samples. AB - Hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type-1 (HIV-1) are transfusion-transmitted human pathogens that have a major impact on blood safety and public health worldwide. We developed a microarray multiplex assay for the simultaneous detection and discrimination of these three viruses. The microarray consists of 16 oligonucleotide probes, immobilized on a silylated glass slide. Amplicons from multiplex PCR were labeled with Cy-5 and hybridized to the microarray. The assay detected 1 International Unit (IU), 10 IU, 20 IU of HBV, HCV, and HIV-1, respectively, in a single multiplex reaction. The assay also detected and discriminated the presence of two or three of these viruses in a single sample. Our data represent a proof-of concept for the possible use of highly sensitive multiplex microarray assay to screen and confirm the presence of these viruses in blood donors and patients. PMID- 17407766 TI - The somatotopic organization of trigeminal premotoneurons in the cat brainstem. AB - This study was performed to complement the results of prior intracellular recording and labeling studies by investigating the general distribution pattern of trigeminal premotoneurons in the cat brainstem using the retrograde tracing methods. The results of the present study reconfirmed the presence of premotoneurons in the trigeminal principal and oral nuclei following horseradish peroxidase injections into the jaw-opening (JO) or jaw-closing (JC) nucleus. Furthermore, we found that labeled cells from the JO nucleus and JC nucleus located in the reticular regions surrounding the trigeminal motor nucleus (Vmo; Vmo shell region) were arranged in a topographic fashion, while those in the parabrachial nucleus, supratrigeminal nucleus, lateral reticular formation caudal to the shell region and raphe nuclei were intermingled with each other. The labeling in the individual nuclei was bilateral with an ipsilateral predominance to each injection site, with the exception of the mesencephalic trigeminal nucleus, where the labeling was ipsilateral to the injection site in the JC nucleus. These results, combined with the data of the previous intracellular tracing studies, indicate that based on the presence of somatotopic organization, premotoneurons can be largely divided into two groups; those projecting to either the JO or the JC nucleus and those projecting to the two nuclei, and we offer the suggestion that roles of premotoneurons for jaw movements differ among the individual nuclei. PMID- 17407767 TI - Changes in NMDA receptor-induced cyclic nucleotide synthesis regulate the age dependent increase in PDE4A expression in primary cortical cultures. AB - NMDA receptor-induced cAMP and cGMP are selectively hydrolyzed by PDE4 and PDE2, respectively, in rat primary cerebral cortical and hippocampal cultures. Because cAMP levels regulate the expression of PDE4 in rat primary cortical cultures, we examined the manner in which NMDA receptor activity regulates the age-dependent increase in the expression of PDE4A observed in vivo and in vitro. Inhibiting the activity of NR2B subunit with ifenprodil blocked NMDA receptor-induced cGMP synthesis and increased NMDA receptor-induced cAMP levels in a manner that reduced PDE4 activity. Therefore, NR1/NR2B receptor-induced cGMP signaling is involved in an acute cross-talk regulation of NR1/NR2A receptor-induced cAMP levels, mediated by PDE4. Chronic inhibition of NMDA receptor activity with MK 801 reduced PDE4A1 and PDE4A5 expression and activity in a time-dependent manner; this effect was reversed by adding the PKA activator dbr-cAMP. Inhibiting GABA receptors with bicuculline increased NMDA receptor-induced cAMP synthesis and PDE4A expression in cultures treated between DIV 16 and DIV 21 but not in cultures treated between DIV 8 and DIV 13. This effect was due to a high tone of NMDA receptor-induced cGMP in younger cultures, which negatively regulated the expression of PDE4A by a PKG-mediated process. The present results are consistent with behavioral data showing that both PDE4 and PDE2 are involved in NMDA receptor-mediated memory processes. PMID- 17407768 TI - Proline induces alterations on nucleotide hydrolysis in synaptosomes from cerebral cortex of rats. AB - In the present study we investigated the in vivo (acute and chronic) and in vitro effects of proline on NTPDase and 5'-nucleotidase activities in synaptosomes obtained from cerebral cortex of rats. For acute administration, 29-day-old rats received one subcutaneous injection of proline (18.2 micromol/g body weight) or an equivalent volume of 0.9% saline solution (control) and were killed 1 h later. For chronic treatment, buffered proline was injected subcutaneously into rats twice a day at 10 h intervals from the 6th to the 28th day of age. Rats were killed 12 h after the last injection. Results showed that acute and chronic proline administration provoked a reduction (25%) of ATP hydrolysis, but did not alter ADP and AMP hydrolysis. We also verified the in vitro effect of proline (3.0 microM-1.0 mM) on nucleotide hydrolysis in synaptosomes from cerebral cortex of rats. In contrast to the in vivo studies, it was not observed any statistically significant alteration on ATP, ADP and AMP hydrolysis. In conclusion, according to our results, it seems reasonable to postulate that proline administration alters the hydrolysis of ATP and probably affects the responses mediated by adenine nucleotides in the central nervous system of proline treated rats. PMID- 17407769 TI - Synthesis of galactose-mimicking 1H-(1,2,3-triazol-1-yl)-mannosides as selective galectin-3 and 9N inhibitors. AB - 1H-[1,2,3]-Triazol-1-yl mannosides have been synthesized as inhibitors for the beta-galactoside-binding family of galectin proteins. Easier synthetic access to C1 in mannose, as compared to C3 in galactose, for attachment of affinity enhancing triazoles rendered a synthetic advantage. The best mannose-derived inhibitor for galectin-9N, 4-benzylaminocarbonyl-1H-[1,2,3]-triazol-1-yl beta-D mannopyranoside, had a Kd value of 540 microM, which compares favorably with its galactoside counterpart (Kd=670 microM) and with LacNAc (Kd=500 microM). PMID- 17407770 TI - Total synthesis of cleistetroside-2, partially acetylated dodecanyl tetrarhamnoside derivative isolated from Cleistopholis patens and Cleistopholis glauca. AB - The total synthesis of a partially acetylated dodecanyl tetrarhamnoside derivative, cleistetroside-2, which was isolated from Cleistopholis patens and Cleistopholis glauca and showed significant in vitro antibacterial activity against the Gram-positive bacteria, was achieved for the first time. PMID- 17407771 TI - From lactose towards a novel galactosylated cyclooctenone. AB - Lactose was converted into a disaccharide precursor incorporating an allyl vinyl ether substructure within the glucose residue by vinylation, regioselective silylation, tosylation and subsequent elimination. The thermal Claisen rearrangement led to a novel galactosylated cyclooctenone exhibiting a boat-chair conformation. PMID- 17407772 TI - Metal complexation of chitosan and its glutaraldehyde cross-linked derivative. AB - The physicochemical characterization of metal complexed with chitosan (CS) and its glutaraldehyde cross-linked derivative (CSGA) was investigated. Seven metal ions from chromium through zinc of the first row of the transition metals were selected for complexation. Structural features pertinent to where and how metals bind into both polymers are our main interest. Studies using solid-state NMR spectroscopy and XRPD (X-ray powder diffraction) supported by ESR spectroscopy, ICP-OES (inductively couple plasma-optical emission spectroscopy) and far-FTIR spectroscopy for metal interaction with nitrogen sites at C-2 of the metal polymer complexes were performed. Theoretical calculations of the metal-polymer ratio, the approximate charges on nitrogen for both amine and imino-linker, and the proton affinity between an alcohol group from the polymer and an amino/imino group are reported. A helical coiled chitosan model and a 2C1L (two-chitosans with one linker) model are proposed here. The metal uptake mechanism for both polymers is concluded to be absorption within the polymers, rather than adsorption on the polymer surface. PMID- 17407773 TI - Synthesis and anti-inflammatory activity of N-phthalimidomethyl 2,3-dideoxy- and 2,3-unsaturated glycosides. AB - 3,4,6-Tri-O-acetyl-D-galactal, 3,4,6-tri-O-acetyl-D-glucal and 3,6,2',3',4'6' hexa-O-acetyl-D-lactal were reacted with N-hydroxymethylphthalimide and boron trifluoride etherate to produce the corresponding phthalimidomethyl unsaturated glycosides via Ferrier rearrangement. When the galactal derivative was used, a non-Ferrier rearrangement product was also isolated as a minor product under classical Ferrier conditions. Phthalimidomethyl deoxy glycosides were readily prepared by hydrogenation of the unsaturated glycosides. Following deacetylation, the anti-inflammatory activities of these compounds were tested on mice and three were found to possess potent activity compared to hydrocortisone sodium succinate (HSS). PMID- 17407774 TI - Synthesis of 4-O-glycosylated 1-deoxynojirimycin derivatives as disaccharide mimics-based inhibitors of human beta-glucocerebrosidase. AB - Examples of a new type of inhibitor of human beta-glucocerebrosidase based on imino-disaccharides as glycosylceramide mimetics have been synthesized by way of the glycosylation of 1-deoxynojirimycin derivatives with 2,3,4,6-tetra-O-acetyl alpha-D-glucopyranosyl bromide. PMID- 17407775 TI - Effective chemoenzymatic synthesis of p-aminophenyl glycosides of sialyl N acetyllactosaminide and analysis of their interactions with lectins. AB - A convenient chemoenzymatic procedure for the synthesis of p-aminophenyl glycosides of sialyl N-acetyllactosaminide has been developed from p-nitrophenyl N-acetyl-beta-D-glucosaminide as starting material through three steps: synthesis of p-nitrophenyl N-acetyllactosaminide with beta-D-galactosidase, chemical reduction of the p-nitrophenyl group, and sialylation with sialyltransferase. The p-aminophenyl glycosides were then successfully biotin-labeled through the coupling with N-(+)-biotinyl-6-aminohexanoic acid to afford biotinylated oligosaccharides with an aminohexanosyl group and phenyl group as the spacers between the biotin and glycan. Furthermore, the biotin-labeled sugars were shown to be useful for immobilization and assay of the carbohydrate-lectin interactions by an optical biosensor based on surface plasmon resonance. PMID- 17407776 TI - Inner speech slips exhibit lexical bias, but not the phonemic similarity effect. AB - Inner speech, that little voice that people often hear inside their heads while thinking, is a form of mental imagery. The properties of inner speech errors can be used to investigate the nature of inner speech, just as overt slips are informative about overt speech production. Overt slips tend to create words (lexical bias) and involve similar exchanging phonemes (phonemic similarity effect). We examined these effects in inner and overt speech via a tongue-twister recitation task. While lexical bias was present in both inner and overt speech errors, the phonemic similarity effect was evident only for overt errors, producing a significant overtness by similarity interaction. We propose that inner speech is impoverished at lower (featural) levels, but robust at higher (phonemic) levels. PMID- 17407777 TI - Neuroprotective effect of L-kynurenine sulfate administered before focal cerebral ischemia in mice and global cerebral ischemia in gerbils. AB - Excessive stimulation of N-methyl-D-aspartate (NMDA) receptors during ischemia contributes to apoptotic and excitotoxic nerve cell death. Kynurenic acid is a selective antagonist at the glycine co-agonist site of the NMDA receptor complex at low concentration, and it is a broad-spectrum excitatory amino acid receptor blocker at high concentration. Kynurenic acid provides neuroprotection in animal models of cerebral ischemia only at very high doses as it hardly crosses the blood-brain barrier. The neuroprotective effect of L-kynurenine sulfate, a precursor of kynurenic acid, was therefore studied because L-kynurenine readily crosses the blood-brain barrier. L-kynurenine sulfate was administered 15 min before permanent focal cerebral ischemia produced by electrocoagulation of the distal middle cerebral artery in mice. L-kynurenine sulfate induced a small decrease in the surface area of the brain infarction (10%, P<0.05) at 30 mg/kg i.p., and it caused strong reductions in infarct size (24-25%, P<0.01) at 100 and 300 mg/kg i.p. Treatment of gerbils with L-kynurenine sulfate at 300 mg/kg i.p. 2 h before a 3-min bilateral carotid occlusion decreased (40%, P<0.01) the pyramidal cell loss in the CA1 area of the hippocampus. Furthermore, L-kynurenine sulfate inhibited the ischemia-induced hypermotility (77%, P<0.001), and decreased (50%, P<0.01) the ischemia-induced deterioration of spontaneous alternation, a measure of spatial memory, without altering the rectal temperature. In conclusion, the administration of L-kynurenine can elevate the brain concentration of kynurenic acid to neuroprotective levels, suggesting the potential clinical usefulness of L-kynurenine for the prevention of neuronal loss. PMID- 17407778 TI - Familial aggregation of human susceptibility to co- and multiple helminth infections in a population from the Poyang Lake region, China. AB - Human helminthiases are common in China, especially in rural areas where sanitation conditions are poor. Co- and multiple infections with helminths are strikingly frequent. A cross-sectional parasitological and questionnaire survey was carried out in a population of 3205 individuals belonging to 498 families from five villages in the Poyang Lake region, Jiangxi Province, China, to assess their helminth infection status and to collect information on risk factors for infection. The prevalences for Ascaris lumbricoides, Schistosoma japonicum and Trichuris trichiura were 30.9%, 15.7% and 47%, respectively. Hookworm infection prevalence was low (0.7%). A significant association was observed between A. lumbricoides and T. trichiura infection, and also between S. japonicum and T. trichiura infection. Variance components analysis was undertaken to investigate the aggregation of S. japonicum and the soil-transmitted helminths, A. lumbricoides and T. trichiura. While A. lumbricoides was found to aggregate only at a household level, T. trichiura was shown to cluster predominantly in families. Both genetic and household effects were found to be important in determining the risk of infection with S. japonicum. Variance components analysis for A. lumbricoides/T. trichiura co-infections indicated a significant domestic environmental effect, attributable for 32.7% of the co-infection risk. Aggregation of S. japonicum/T. trichiura co-infection was also observed at a household level. The risk of infection with multiple helminth species, although mainly environmentally influenced, was also shown to have significant involvement of genetic and household components. The results of this study indicate that a shared household is a major contributing risk factor for helminth co-infections and emphasises the need for increased standards of sanitation and hygiene to prevent parasite transmission. Further, the results suggest that susceptibility to one helminth infection is not completely independent of another, and that there exist common genetic factors underlying infection with multiple helminth species. PMID- 17407779 TI - Microwave-Accelerated Surface Plasmon-Coupled Directional Luminescence: application to fast and sensitive assays in buffer, human serum and whole blood. AB - The applicability of a new technique, Microwave-Accelerated Surface Plasmon Coupled Luminescence (MA-SPCL) for fast and sensitive bioassays in buffer, serum and whole blood using quantum dots as luminescence reporters is demonstrated. In this regard, a model bioassay based on the well-known interactions of biotin and streptavidin is used. Using MA-SPCL, the bioassay was kinetically completed within 1 min with the use of low power microwave heating as compared to the identical bioassay which took in excess of 30 min to reach >95% completion at room temperature, a 30-fold increase in assay kinetics. The luminescence emission from the quantum dots was coupled to surface plasmons of the gold film, enabling the detection of the luminescence emission in a highly directional fashion as compared to the normal isotropic emission, for enhanced sensitivity and detection. The combined effect of microwaves for faster assay kinetics, with surface plasmon-coupled luminescence for sensitive luminescence measurements, has also made possible the demonstration of the use of the MA-SPCL technique for assays run in complex media, such as human serum and whole blood, where the same assay could not be performed at room temperature due to the coagulation of blood. In the MA-SPCL assay run in serum and whole blood, the luminescence intensity from 33 nM quantum dots was 75% and 20% that of the luminescence intensity from the assay run in buffer, with a signal to noise ratio of 12.5 and 3, respectively. PMID- 17407780 TI - Impaired p38 MAPK/HSP27 signaling underlies aging-related failure in opioid mediated cardioprotection. AB - Cardioprotection and preconditioning mediated via G-protein-coupled receptors may be lost or impaired with advancing age, limiting ischemic tolerance and the ability to pharmacologically protect older hearts from ischemic injury. Our preliminary findings indicated a loss of delta-opioid receptor-mediated protection in aged vs. young mouse hearts, which may involve alterations in protective kinase signaling. In the present study, we tested the hypothesis that aging-related loss of opioid-triggered cardioprotection involves failure to activate p38 MAPK and its distal signaling targets. Langendorff-perfused hearts from young (10-14 weeks) or aged (24-26 months) C57 mice underwent 25-min ischemia and 45-min reperfusion in the presence or absence of 1 micromol/l DPDPE (delta-opioid agonist) or 1 micromol/l anisomycin (activator of p38 MAPK), and functional recovery and protein activation/phosphorylation were assessed. Contractile recovery was similar in untreated young and aged hearts (50+/-2% and 53+/-5%, respectively), and was enhanced by DPDPE in young hearts only (67+/-3%). Immunoblot analysis revealed that DPDPE comparably activated or phosphorylated GRK2, Akt, ERK1/2 and p70S6 kinase in young and aged hearts, whereas aging abrogated the stimulatory effects of DPDPE on p38 MAPK and HSP27. Treatment with anisomycin elicited comparable activation of p38 MAPK and HSP27 in both young and aged hearts, coupled with a pronounced and equivalent cardioprotection in the two groups (73+/-3% and 77+/-2%, respectively), an effect abolished by the p38 MAPK inhibitor, SB203580. These data indicate that aging-related loss of delta-opioid mediated cardioprotection involves failure to activate p38 MAPK and HSP27. Direct targeting of this pathway elicits comparable protection in both age groups. PMID- 17407781 TI - Binding of ATP to TK1-like enzymes is associated with a conformational change in the quaternary structure. AB - Human thymidine kinase 1 (hTK1) and structurally related TKs from other organisms catalyze the initial phosphorylation step in the thymidine salvage pathway. Though ATP is known to be the preferred phosphoryl donor for TK1-like enzymes, its exact binding mode and effect on the oligomeric state has not been analyzed. Here we report the structures of hTK1 and of the Thermotoga maritima thymidine kinase (TmTK) in complex with the bisubstrate inhibitor TP4A. The TmTK-TP4A structure reveals that the adenosine moiety of ATP binds at the subunit interface of the homotetrameric enzyme and that the majority of the ATP-enzyme interactions occur between the phosphate groups and the P-loop. In the hTK1 structure the adenosine group of TP4A exhibited no electron density. This difference between hTK1 and TmTK is rationalized by a difference in the conformation of their quaternary structure. A more open conformation, as seen in the TmTK-TP4A complex structure, is required to provide space for the adenosine moiety. Our analysis supports the formation of an analogous open conformation in hTK1 upon ATP binding. PMID- 17407782 TI - The extracellular chaperone clusterin potently inhibits human lysozyme amyloid formation by interacting with prefibrillar species. AB - We have studied the effects of the extracellular molecular chaperone, clusterin, on the in vitro aggregation of mutational variants of human lysozyme, including one associated with familial amyloid disease. The aggregation of the amyloidogenic variant I56T is inhibited significantly at clusterin to lysozyme ratios as low as 1:80 (i.e. one clusterin molecule per 80 lysozyme molecules). Experiments indicate that under the conditions where inhibition of aggregation occurs, clusterin does not bind detectably to the native or fibrillar states of lysozyme, or to the monomeric transient intermediate known to be a key species in the aggregation reaction. Rather, it seems to interact with oligomeric species that are present at low concentrations during the lag (nucleation) phase of the aggregation reaction. This behavior suggests that clusterin, and perhaps other extracellular chaperones, could have a key role in curtailing the potentially pathogenic effects of the misfolding and aggregation of proteins that, like lysozyme, are secreted into the extracellular environment. PMID- 17407783 TI - Persistence of hyperbaric oxygen-induced oxidative effects after exposure in rat brain cortex tissue. AB - Hyperbaric oxygen (HBO) causes oxidative stress in several organs and tissues. Due to its high rate of blood flow and oxygen consumption, the brain is one of the most sensitive organs to this effect. Many studies have reported oxidative effects of HBO, but there is no comprehensive data about how long this effect persists. The aim of this study was to elucidate the duration of HBO-induced oxidative/antioxidant action. Male Sprague-Dawley rats were divided into 5 groups. Except for the controls, the animals were subjected to 100% oxygen for 2 h at 3 atm and differed from each other by the time to dissection after exposure that began at 30, 60, 90, or 120 min. Thiobarbituric acid-reactive substances (TBARS), as well as superoxide dismutase (SOD) and glutathione peroxidase (GSH Px) activity was determined in brain cortex tissue. Additionally, nitrite-nitrate (NO(x)) concentrations were measured. All measured parameters were found to be significantly increased 30 min after exposure. SOD and GSH-Px levels persisted at significantly high levels for 60 min. In conclusion, the oxidative effect of HBO was shown to persist only for 1 h. Further studies should be performed to elucidate the possible molecular interactions during this period. PMID- 17407784 TI - Adenoviruses increase endothelial cell proliferation, migration, and tube formation: partial reversal by the focal adhesion kinase inhibitor, FRNK. AB - During the course of examining the feasibility of using an adenoviral vector to deliver a potential anti-angiogenic agent to endothelial cells, we discovered that adenoviruses, themselves, have pro-angiogenic activities. Thus, an adenoviral vector containing a green fluorescent protein transgene (Ad-GFP) stimulated the growth, migration, tube formation, and phosphorylation of focal adhesion kinase (FAK) of human lung microvascular endothelial cells. However, adenovirus-mediated endothelial cell mitogenesis, tube formation, and FAK phosphorylation were completely reduced and migration was partially reversed by the addition of a Fak-Related Non-Kinase (FRNK) transgene to the vector. Because FRNK inhibits focal adhesion kinase (FAK) activity, this suggests that the adenoviral effects on endothelial cells are in part mediated through FAK. These data, as well as data obtained in other laboratories, suggest that adenoviruses should be used with caution in cancer gene therapy due to potential pro angiogenic effects. However, some of these untoward effects may be modulated by concurrent use of a FAK inhibitor. PMID- 17407786 TI - Vitamin A and contaminant concentrations in surf scoters (Melanitta perspicillata) wintering on the Pacific coast of British Columbia, Canada. AB - Surf scoters are part of a community of sea ducks on the western coast of North America that have shown signs of long-term, unexplained declines in breeding bird numbers. Substantial numbers of scoters winter in the major harbours on the west coast, after breeding in the west-central northern boreal forest. To address the potential for contaminants to impact the health and survival of those birds, we investigated the condition and contamination of surf scoters during the winters of 1998-2001 at four foraging locations in the Strait of Georgia region of the Pacific coast of Canada. Vitamin A status was evaluated in liver and plasma samples collected from adults and juveniles, as part of a larger assessment of tissue contamination, body condition and biomarker responses. Individuals collected from a relatively contaminated site, Howe Sound, showed consistently low hepatic concentrations of retinol and retinyl palmitate forms of vitamin A, and gender-specific associations of retinyl palmitate with hepatic EROD activity. The relationship of hepatic retinol to retinyl palmitate was not constant across geographic locations, and a clear, linear relationship between the two forms of vitamin A was only evident in birds from the relatively uncontaminated site. This study also identified strong positive relationships between vitamin A and tissue burdens of cadmium and zinc. The positive association between hepatic retinyl palmitate and renal cadmium is similar to one observed in laboratory rats, in which a mechanism of interference with the controlled release of retinol from the liver was suggested. PMID- 17407785 TI - AL1-dependent repression of transcription enhances expression of Tomato golden mosaic virus AL2 and AL3. AB - Studies using Nicotiana benthamiana protoplasts have determined that repression of upstream transcription by AL1 protein enhances AL2 and AL3 expression in Tomato golden mosaic virus (TGMV). Mutations resulting in the inability of TGMV AL1 protein to associate with its cognate binding site, result in a decrease in both AL2 and AL3 expression. Reduced expression correlates with an increase in transcription from the AL62 start site, and decreased transcription from downstream initiation sites (AL1935 and AL1629) present within the AL1 coding region. The results demonstrate that, in a tobacco protoplast system, repression of AL62 transcription, regulated through binding of AL1 protein to sequences in the origin of replication, is required prior to AL2 and AL3 gene expression from the AL1935 and AL1629 viral transcripts. This provides a mechanism to regulate expression of AL2, which is involved in suppression of host defense responses and is required for late gene expression. PMID- 17407787 TI - The characteristics of rhizosphere microbes associated with plants in arsenic contaminated soils from cattle dip sites. AB - Soil microorganisms and plants were studied in samples of arsenic-contaminated soil from two cattle dip sites. The aim was to delineate the parameters that will determine the feasibility of future remediation by growing arsenic-accumulating plants, including the identity and characteristics of some rhizosphere soil microbes. The soil samples contained high total, but low soluble arsenic concentrations which, together with other properties, resembled the previously reported characteristics of dip-site soils from this region of rural Australia. A glasshouse trial demonstrated that dip-site rhizosphere microbes promoted arsenic accumulation by the grass Agrostis tenuis on contaminated dip-site soil without inhibition of growth. The arsenic content of the shoots was increased by 45%. We studied the colonization of roots of dip-site plants by mycorrhizal fungi and tentatively identified six genera of other fungi present in the soil samples. Two plant species growing at the sites, Kikuyu grass (the most abundant plant) and Rainbow fern, exhibited mixed infections of their roots by endomycorrhizal fungi (tentatively identified as Acaulospora and Gigaspora) and by soil-born pathogens. Five rhizosphere bacteria were identified to genus level and we determined the effect of arsenic on their growth. The two most prevalent strains differed greatly in their growth sensitivity to arsenate; Arthrobacter sp. being the most sensitive while Ochrobactrum sp. exhibited exceptional resistance to arsenate. Of the other, less prevalent strains, two were Bacillus spp. and the last, Serratia sp., was the most resistant to arsenite. These findings show the importance of understanding plant-soil microbe interactions for developing future strategies aimed at a phytoremediation-based approach to removing arsenic from soil at dip sites. PMID- 17407788 TI - Prothrombin complex concentrate (Octaplex) in patients requiring immediate reversal of oral anticoagulation. AB - INTRODUCTION: Rapid reversal of anticoagulant effect from the use of vitamin K antagonists (VKA) is essential when acute bleeding or emergency surgery occurs. Prothrombin complex concentrates (PCCs) produce a more rapid effect with a better clinical outcome, and do not cause volume overload as compared with fresh-frozen plasma (FFP). Octaplex is a modern, double virus safeguarded PCC with balanced content of vitamin K-dependent coagulation factors, which ensures fast onset of action and efficacious treatment, i.e. rapid correction of international normalized ratio (INR). MATERIALS AND METHODS: The main purpose of this study was to demonstrate that Octaplex, when individually dosed, efficiently corrects INR to pre-determined levels in patients under oral anticoagulation who have bleeding complications or are undergoing invasive procedures. To measure the efficacy response, the INR achieved after PCC application per patient was calculated as geometric mean of three measurements within 1 h post-infusion. RESULTS: Sixty patients received a median total Octaplex dose of 41.1 (15.3-83.3) IU/kg body weight (bw). Of 56 patients evaluable in terms of efficacy, 51 (91%) showed a response as pre-defined in the protocol and in 52 (93%) the INR decreased to a value below 1.4 within one hour after dosing. The median INR declined from 2.8 (1.5-9.5) to 1.1 (1.0-1.9) within 10 min. All prothrombin complex coagulation factors recovered in parallel. Three patients had minor adverse drug reactions. One patient showed a non-symptomatic parvovirus B19 seroconversion. No thrombotic side effects were observed. CONCLUSIONS: Octaplex is efficacious and safe in immediate correction of dosage-dependent INR in patients with VKA-related deficiency of prothrombin complex coagulation factors. PMID- 17407789 TI - Enhanced fibrinolysis in fingerstick blood samples: a possible role for matrix metalloproteinase-9. PMID- 17407790 TI - Sexually dimorphic effects of acute nicotine administration on arousal and visual spatial ability in non-smoking human volunteers. AB - The effect of an acute administration of nicotine on arousal and visual-spatial ability in healthy non-smoking participants was investigated. Healthy adult volunteers with a mean age of 19.98 years received a transdermal nicotine or placebo patch prior to completing a water-level task and two mental rotation tasks while concurrent psychophysiological recordings were taken. Nicotine administration showed a sexually dimorphic effect on arousal (skin conductance level and heart rate). Evidence of superior performance in males compared to females was found in reaction time and accuracy measures for the visual-spatial tasks. However, performance reflected the interaction between sex and nicotine. Nicotine slowed reaction times in the mental rotation tasks more extensively in females than males. Nicotine also reduced confidence in performance during the water-level task in males, but not in females. The effects of nicotine on visual spatial ability may reflect the interactive effects of sex and changes in arousal levels induced by nicotine administration. PMID- 17407791 TI - Baclofen alters flash-evoked potentials in Long-Evans rats. AB - This experiment examined the effects of the GABA-B agonist baclofen on flash evoked potentials (FEPs) recorded from both the visual cortex (VC) and superior colliculus (SC) of chronically implanted male Long-Evans rats. FEPs were recorded at 5, 25, 45, and 65 min following intraperitoneal injections of saline, and of 1.25, 2.5, 5.0, and 10.0 mg/kg baclofen on separate days. In the VC, the amplitude of components P(23), P(37), N(55), N(150), and P(242) increased, while the amplitude of components N(31) and P(48) decreased following baclofen administration. P(88) was unchanged. In the SC, components P(28), N(49), N(55), and N(59) were reduced in amplitude, while P(39) was unaffected by baclofen. These effects on amplitudes were dose- and time-dependent. Many peak latencies in the VC and SC were altered by baclofen, although there was no obvious pattern of change, with some decreasing, a few increasing, and others unchanged. Body temperature was recorded in a separate group of animals, with both the 5.0 and 10.0 mg/kg doses of baclofen producing significant hypothermia. The 10.0 mg/kg dose of baclofen resulted in a significant decrease in movement during the recording sessions, but not in subsequent open field observations. The results show the involvement of GABA-B receptors in the production/modulation of the various components of FEPs. PMID- 17407792 TI - Parent-of-origin effect and genomic imprinting of the HTR2A receptor gene T102C polymorphism in psychosis. AB - Evidence that HTR2A receptor gene may be subject to genomic imprinting prompted us to examine a collection of family trios for evidence of an association between the HTR2A T102C polymorphism and psychosis in schizophrenia or bipolar disorder. We also tested for the possibility of imprinting by employing quantitative RT-PCR to measure the relative expression of post-mortem brain mRNA for each allele in 45 subjects who were heterozygous for the T102C polymorphism. We found that the ratio of C102 to 102T allele mRNA expression was the same in major psychoses and healthy controls. There was no genetic association between HTR2A T102C with either schizophrenia or bipolar disorder under the assumption of a parent-of origin effect, and these data together essentially exclude imprinting at this locus as a potential explanation for the complex inheritance observed in major psychoses. PMID- 17407793 TI - Methyl-CpG-binding domain (MBD) proteins in plants. AB - Cytosine methylation is the most prevalent epigenetic modification of plant nuclear DNA, which occurs in symmetrical CpG or CpNpG as well as in non symmetrical contexts. Intensive studies demonstrated the central role played by cytosine methylation in genome organization, gene expression and in plant growth and development. However, the way by which the methyl group is interpreted into a functional state has only recently begun to be explored with the isolation and characterization of methylated DNA binding proteins capable of binding 5 methylcytosine. These proteins belong to an evolutionary conserved protein family initially described in animals termed methyl-CpG-binding domain (MBD) proteins. Here, we highlight recent advances and present new prospects concerning plant MBD proteins and their possible role in controlling chromatin structure mediated by CpG methylation. PMID- 17407794 TI - Clinical and angiographic features of large left main coronary artery aneurysms. AB - Coronary artery aneurysms are defined as coronary dilatations which exceed the diameter of normal adjacent segments by 1.5 times. Left main coronary artery (LMCA) is the least frequently involved artery with a prevalence of 0.1%. Majority of coronary artery aneurysms are atherosclerotic in origin. Their size and clinical presentation is variable. A number of complications have been reported to occur during the course of the disease including thrombosis and distal embolization, myocardial ischemia and/or infarction, dissection, vasospasm, calcification, fistulization and very rarely rupture. Large aneurysms of LMCA represent a potentially fatal condition even without concomitant atherosclerotic coronary disease. Because of rarity of coronary artery aneurysms, it is difficult to standardize treatment. In this article, we presented 4 cases of large LMCA aneurysms with various clinical and angiographic features. PMID- 17407795 TI - Sustained benefit of tadalafil in patients with pulmonary arterial hypertension with prior response to sildenafil: a case series of 12 patients. PMID- 17407796 TI - On the waiting list for possible coronary revascularisation. Symptoms relief during the first year and association between quality of life and the very long term mortality risk. AB - AIM: To describe: a/ the improvement in quality of life (QoL) among patients on the waiting list for coronary revascularisation and b/ the association between QoL and very long-term mortality. PATIENTS: All patients on the waiting list for possible coronary revascularisation in western Sweden during one week in September 1990. METHODS: QoL was assessed at the start of the survey and one year later among patients who both were and were not revascularised. Survival data were gathered for the subsequent 14 years. RESULTS: From the start, 883 patients were evaluated in the survey. Among patients who were revascularised, an improvement was seen in all the aspects of QoL that were studied during the first year as compared with patients who were not revascularised, in whom only minor changes in QoL were seen during the first year. After one year, there were seven aspects of QoL which were significantly associated with the risk of death during the subsequent 14 years, when adjusting for age, sex, previous history and extent of coronary artery disease. They were: tiredness (OR=1.4), weakness (OR=1.5), lack of energy (OR=1.5), inability to react (OR=1.7), use of sedatives (OR=3.2), dyspnea when dressing (OR=2.1) and chest pain when dressing (OR=1.9). CONCLUSION: Among patients on the waiting list for possible coronary revascularisation, there was a marked improvement in QoL among those who were revascularised. In a variety of aspects of QoL, an association with the very long-term risk of death was observed. PMID- 17407797 TI - Bacterial diversity of soils assessed by DGGE, T-RFLP and SSCP fingerprints of PCR-amplified 16S rRNA gene fragments: do the different methods provide similar results? AB - Bacterial communities of four arable soils--pelosol, gley, para brown soil, and podsol brown soil--were analysed by fingerprinting of 16S rRNA gene fragments amplified from total DNA of four replicate samples for each soil type. Fingerprints were generated in parallel by denaturing gradient gel electrophoresis (DGGE), terminal restriction fragment length polymorphism (T RFLP), and single strand conformation polymorphism (SSCP) to test whether these commonly applied techniques are interchangeable. PCR amplicons could be separated with all three methods resulting in complex ribotype patterns. Although the fragments amplified comprised different variable regions and lengths, DGGE, T RFLP and SSCP analyses led to similar findings: (a) a clustering of fingerprints which correlated with soil physico-chemical properties, (b) little variability between the four replicates of the same soil, (c) the patterns of the two brown soils were more similar to each other than to those of the other two soils, and (d) the fingerprints of the different soil types revealed significant differences in a permutation test, which was recently developed for this purpose. PMID- 17407798 TI - Growth characteristics of microorganisms on commercially available animal-free alternatives to tryptic soy medium. AB - The growth characteristics of a range of bacteria and fungi that are routinely used in quality control practices were compared for two representative vegetable based tryptic soy formulations. All of the representative microorganisms grew well on the vegetable-based media and the media provided suitable recoveries of the organisms following simulated storage. Subtle phenotypic changes were observed between cultures grown on different media, but these did not significantly change the strain identification. PMID- 17407799 TI - Activity ratios of 137Cs, 90Sr and 239+240Pu in environmental samples. AB - Both global and Chernobyl fallout have resulted in environmental contamination with radionuclides such as 137Cs, 90Sr and 239+240Pu. In environmental samples, 137Cs and 239+240Pu can be divided into the contributions of either source, if also the isotopes 134Cs and 238Pu are measurable, based on the known isotopic ratios in global and Chernobyl fallout. No analogous method is available for 90Sr. The activity ratios of Sr to Cs and Pu, respectively, are known for the actual fallout mainly from air filter measurements; but due to the high mobility of Sr in the environment, compared to Cs and Pu, these ratios generally do not hold for the inventory many years after deposition. In this paper we suggest a method to identify the mean contributions of global and Chernobyl fallout to total Sr in soil, sediment and cryoconite samples from Alpine and pre-Alpine regions of Austria, based on a statistical evaluation of Sr/Cs/Pu radionuclide activity ratios. Results are given for Sr:Cs, Sr:Pu and Cs:Pu ratios. Comparison with fallout data shows a strong depletion of Sr against Cs and Pu. PMID- 17407800 TI - Interferon-gamma induced disruption of GABAergic inhibition in the spinal dorsal horn in vivo. AB - The proinflammatory cytokine interferon-gamma (IFN-gamma), which can be present in elevated levels in the central nervous system during pathological conditions, may be involved in the generation of persistent pain states by inducing neuronal hyperexcitability. The aim of the present study was to examine whether loss of dorsal horn GABAergic inhibition may underlie this IFN-gamma-mediated neuronal hyperexcitability. Repetitive intrathecal injections of recombinant rat IFN-gamma (1000 U) or control buffer were administered to rats every second day for eight days. Electrophysiological recordings from lumbar dorsal horn neurons (n=46) were performed under halothane anaesthesia. Cellular responses were recorded before, during and after microiontophoretic application of the GABA antagonist bicuculline. In control animals, all cellular responses studied were significantly enhanced in the presence of bicuculline, including increased spontaneous activity, enhanced responses to innocuous and noxious mechanical stimulation and reduced paired-pulse depression. In contrast, in IFN-gamma treated animals, bicuculline ejection had little or no facilitating effect on neuronal responses and instead a significant proportion of neurons displayed reduced responses. Seventy-four percent of cells from IFN-gamma treated animals showed a reduction in the response to noxious stimulation and 47% of the cells showed increased rather than reduced paired-pulse depression in the presence of bicuculline, thus suggesting IFN-gamma-induced excitatory actions by GABA. These findings show that the prolonged presence of increased levels of IFN-gamma in the central nervous system may contribute to the generation of central sensitization and persistent pain by reducing inhibitory tone in the dorsal horn. This implies a potential link between disinhibition and cytokine action in the spinal cord. PMID- 17407801 TI - Comment on Diatchenko et al. Catechol-O-methyltransferase gene polymorphisms are associated with multiple pain-evoking stimuli. Pain 2006;125:216-24. PMID- 17407802 TI - Alcohol use as a behavioural sign of disinhibition: evidence from J.A. Gray's model of personality. AB - Based on Gray's Reinforcement Sensitivity Theory, the influence of behavioural disinhibition upon alcohol consumption was studied. A sample of undergraduates answered different questionnaires related to the Behavioural Inhibition System and Behavioural Activation System. In relation to alcohol use, three aspects of alcohol consumption were assessed: frequency, quantity of alcohol intake and the age at first drink. From a series of correlation and regression analyses, we found that both high scores on BAS-related scales and low scores on those scales related to the BIS were jointly associated with current alcohol-taking habits. Additionally, the Sensitivity to Reward (SR) scale (BAS) was negatively correlated with, and a predictor of, the onset age of alcohol use. We conclude by proposing that research on alcohol use can benefit from this well-grounded theory of the neuropsychology of the individual differences. PMID- 17407803 TI - Androgen therapy in women: what we think we know. AB - To date there is no clinical indication for the use of testosterone in women and long-term safety data is lacking. This minireview examines androgen therapy in women. Issues relating to research into sexual function and androgens in women are discussed. Androgen physiology is described and studies examining endogenous androgens in women cited. A Cochrane review found that adding testosterone to a hormone therapy regimen has beneficial effects on sexual function in postmenopausal women and subsequent studies have supported the role of testosterone in sexual function and well-being. However studies on safety and efficacy of testosterone beyond 24 weeks are lacking. PMID- 17407804 TI - Differential effects of typical and atypical antipsychotics on brain myelination in schizophrenia. AB - CONTEXT: Imaging and post-mortem studies provide converging evidence that patients with schizophrenia have a dysregulated developmental trajectory of frontal lobe myelination even in adulthood. Atypical antipsychotics have been shown to have a wide spectrum of efficacy across multiple psychiatric diseases and to be particularly efficacious in treatment resistant cases of disorders such as schizophrenia. OBJECTIVE: To test the a priori hypothesis that antipsychotic medications may differentially impact frontal lobe myelination in patients with schizophrenia. DESIGN, SETTING, AND PARTICIPANTS: Participants ranged in age from 18-35 years, were all male, and were recruited by a single group of investigators using the same criteria. Two cohorts of subjects with schizophrenia early in their disease who were treated either with oral risperidone (Ris) or fluphenazine decanoate (Fd) were imaged in conjunction with cohorts of healthy controls. Each cohort was imaged using a different MRI instrument using identical imaging sequences. MAIN OUTCOME MEASURE: MRI measures of frontal lobe white matter volume. RESULTS: We estimated differences due to differences in the MRI instruments used in the two studies in the two healthy control groups matched to the patient samples, adjusting for age and other covariates. We then statistically removed those differences (which we assumed were due to instrument effects) from the data in the schizophrenia samples by subtraction. Relative to the differences seen in controls, the two groups of schizophrenic patients differed in their pattern of frontal lobe structure with the Ris-treated group having significantly larger white matter volume than the Fd group. CONCLUSIONS: The results suggest that the choice of antipsychotic treatment may differentially impact brain myelination in adults with schizophrenia. Prospective studies are needed to confirm this finding. MRI can be used to dissect subtle differences in brain tissue characteristics and thus could help clarify the effect of pharmacologic treatments on neurodevelopmental and pathologic processes in vivo. PMID- 17407805 TI - No association evidence between schizophrenia and dystrobrevin-binding protein 1 (DTNBP1) in Taiwanese families. AB - Several linkage studies have shown significant linkage of schizophrenia to chromosome 6p region, which includes the positional candidate genes, Dystrobrevin binding protein 1 (DTNBP1). The aim was to examine the association evidence of the candidate gene in 693 Taiwanese families with at least two affected siblings of schizophrenia. We genotyped nine SNPs of this gene with average intermarker distance of 17 kb. Intermarker linkage disequilibrium was calculated with GOLD. Single locus and haplotype association analyses were performed with TRANSMIT program. We found no significant association between schizophrenia and DTNBP1 either through single locus or haplotype analyses. We failed to replicate the association evidence between DTNBP1 and schizophrenia and this gene may not play a major role in the etiology of schizophrenia in this Taiwanese family sample. PMID- 17407806 TI - Metacognition within narratives of schizophrenia: associations with multiple domains of neurocognition. AB - Research has suggested many with schizophrenia experience impairments in metacognition, or difficulties apprehending their own thoughts and the thoughts of others, and that those deficits are not reducible to a single symptom or cognitive impairment. While links between metacognition and more severe levels of symptoms have emerged, less clear is whether there are consistent associations between metacognition and other neurocognitive capacities. Accordingly the current study sought to examine whether different patterns of metacognition deficits have different neurocognitive correlates. Narratives were gathered from 69 adults with schizophrenia spectrum disorder using the Indiana Psychiatric Illness Interview along with a symptom interview and neurocognitive battery including subtests of the Wechsler Adult Intelligence Scale III, Wechsler Memory Scale III and the Wisconsin Card Sorting Test. Metacognitive capacity within the narrative interview was assessed using the Metacognition Assessment Scale and participants were divided based on those scores into three groups: minimal self reflectivity/not decentered (n=25); basic self-reflectivity/not decentered (n=33); and basic self-reflectivity/decentered (n=11). Basic self-reflectivity refers to the ability to distinguish one's own thoughts and feelings while decentered refers to the ability to see others as having independent perspectives and relationships with one another. MANOVA and ANOVA comparing groups revealed that the participants lacking basic self-reflectivity had significantly poorer working memory and more symptoms of disorganization, while participants able to see others as having independent perspectives and relationships demonstrated better visual memory. Results suggest different deficits in metacognition may be linked to different neurocognitive capacities. PMID- 17407807 TI - Evidence that the inhibitory effects of guanidinoacetate on the activities of the respiratory chain, Na+,K+-ATPase and creatine kinase can be differentially prevented by taurine and vitamins E and C administration in rat striatum in vivo. AB - Guanidinoacetate methyltransferase (GAMT) deficiency is an inherited neurometabolic disorder biochemically characterized by tissue accumulation of guanidinoacetate (GAA) and depletion of creatine. Affected patients present epilepsy and mental retardation whose etiopathogeny is unclear. In a previous study we showed that instrastriatal administration of GAA caused a reduction of Na(+),K(+)-ATPase and creatine kinase (CK) activities, as well as an increase in TBARS (an index of lipid peroxidation). In the present study we investigated the in vitro and in vivo effects of GAA on glucose uptake from [U-(14)C] acetate (citric acid cycle activity) and on the activities of complexes II, II-III, III and IV of the respiratory chain in striatum of rats. Results showed that 50 and 100 microM GAA (in vitro studies) and GAA administration (in vivo studies) significantly inhibited complexes II and II-III, respectively, but did not alter complexes III and IV, as well as CO(2) production. We also studied the influence of taurine or vitamins E and C on the inhibitory effects caused by intrastriatal administration of GAA on complexes II and II-III, Na(+),K(+)-ATPase and CK activities, and on TBARS in rat striatum. Pre-treatment with taurine and vitamins E and C revealed that taurine prevents the effects of intrastriatal administration of GAA on the inhibition of complex II, complex II-III, and Na(+),K(+)-ATPase activities. Vitamins E and C prevent the effects of intrastriatal administration of GAA on the inhibition of CK and Na(+),K(+)-ATPase activities, and on the increase of TBARS. The data suggest that GAA in vivo and in vitro treatment disturbs important parameters of striatum energy metabolism and that oxidative damage may be mediating these effects. It is presumed that defects in striatum bioenergetics might be involved in the pathophysiology of striatum damage characteristic of patients with GAMT-deficiency. PMID- 17407809 TI - Solid lipid microparticles containing the sunscreen agent, octyl dimethylaminobenzoate: effect of the vehicle. AB - Solid lipid microparticles (SLMs) loaded with the sunscreen agent, octyl dimethylaminobenzoate (ODAB), were prepared in order to achieve enhanced sunscreen photostability. The microparticles were produced by the melt dispersion technique using glyceryl behenate as lipidic material and poloxamer 188 as the emulsifier. The obtained SLMs showed proper features in terms of morphology, size distribution (1.67-15.81 microm) and ODAB loading (16.15+/-0.11%, w/w). The sunscreen release from the SLMs was slower than its dissolution rate and the photodecomposition of ODAB was markedly decreased (>51.3%) by encapsulation into the lipid microparticles. The efficacy of the SLM carrier system was also evaluated after their introduction in model topical formulations (i.e., hydrogel and oil-in-water emulsion). Further in vitro release measurements, performed using Franz diffusion cells with polycarbonate membranes, indicated that the retention capacity of the microparticles was lost after their incorporation into the emulsion, whereas it was retained in the hydrogel. Moreover, the SLMs achieved a reduction of the sunscreen photodegradation in the hydrogel vehicle (the ODAB loss decreased from 87.4% to 59.1%), whereas no significant photoprotective effect was observed in the emulsion. Therefore, the efficacy of the ODAB-loaded SLMs was markedly affected by the vehicle. PMID- 17407808 TI - Improvement in memory and static balance with abstinence in alcoholic men and women: selective relations with change in brain structure. AB - We investigated whether changes in memory or static balance in chronic alcoholics, occurring with abstinence or relapse, are associated with changes in lateral and fourth ventricular volume. Alcoholics meeting DSM-IV criteria for Alcohol Dependence (n=15) and non-alcoholic controls (n=26) were examined twice at a mean interval of 2 years with standard Wechsler Abbreviated Scale of Intelligence (WASI), Wechsler Memory Scale-Revised (WMS-R) tests, an ataxia battery, and structural MRI. At study entry, alcoholics had been abstinent on average for over 4 months and achieved lower scores than controls on WASI General IQ Index, WMS-R General Memory Index, and the ataxia battery. The 10 alcoholics who maintained sobriety at retest did not differ at study entry in socio demographic measures, alcohol use, or WASI and WMS-R summary scores from the five relapsers. At follow-up, abstainers improved more than controls on the WMS-R General Memory Index. Ataxia tended to improve in abstainers relative to controls. Associations were observed between memory and lateral ventricular volume change and between ataxia and fourth ventricular volume change in alcoholics but not in the controls. Both memory and ataxia can improve with sustained sobriety, and brain-behavior associations suggest selective brain structural substrates for the changes observed. PMID- 17407810 TI - Porous hydroxyapatite tablets as carriers for low-dosed drugs. AB - The present study evaluated an innovative technique for the manufacturing of low dosed tablets. Tablets containing hydroxyapatite and a pore forming agent (50% (w/w) Avicel PH 200/20, 37.5% and 50% corn starch/37.5% sorbitol) were manufactured by direct compression followed by sintering. The influence of pore forming agent (type and concentration), sinter temperature and sinter time on tablet properties was investigated. Sintering (1250 degrees C) revealed tablets with an acceptable friability (<1%). Using 50% (w/w) Avicel PH 200 as pore forming agent resulted in tablets combining the highest porosity (50%) and the highest median pore diameter (5 microm). Aqueous drug solutions (metoprolol tartrate, riboflavin sodium phosphate) were spiked on the tablet surface. The maximum volume of drug solution absorbed was limited (2x100 microl), revealing that these porous carriers were ideal for low dosed formulations. Drug release from the tablets was slow, independent of the drug. To accelerate drug release, tablets were manufactured using a modified gelcasting technique yielding tablets with a median pore size of 60 and 80 microm. Release from these tablets was drastically increased indicating that the permeability of the tablets was influenced by the pore size, shape and connectivity of the porous network. Changing and controlling these parameters made it possible to obtain drug delivery systems providing different drug delivery behaviour. PMID- 17407811 TI - Novel pyridyl-fused 3-amino chroman derivatives with dual action at serotonin transporter and 5-HT1A receptor. AB - Structural modifications of the initial lead, 3-aminochroman (4), led to the identification of a novel series of pyridyl-fused amino chroman derivatives (5-8) and the structural isomers (9-12). The compounds described were evaluated for dual 5-HT transporter inhibitory and 5-HT(1A) receptor activities. The design strategy, synthesis, and in vitro biological characterization for these novel compounds are described. PMID- 17407812 TI - PTP1B inhibitor Ertiprotafib is also a potent inhibitor of IkappaB kinase beta (IKK-beta). AB - Ertiprotafib was developed as an inhibitor of PTP1B for the treatment of type 2 diabetes. It normalized the plasma glucose and insulin levels in diabetic animal models, and progressed to a phase II clinical trial. Multiple in vivo targets of Ertiprotafib, in addition to PTP1B inhibition, have been suggested. In this study, Ertiprotafib was also shown to be a potent inhibitor of IkappaB kinase beta (IKK-beta), with an IC(50) of 400nM. PMID- 17407813 TI - Synthesis and antitubercular activity of phenothiazines with reduced binding to dopamine and serotonin receptors. AB - Analogs of the psychotropic phenothiazines were synthesized and examined as antitubercular agents against Mycobacterium tuberculosis H37Rv. The compounds were subsequently counter-screened for binding to the dopaminergic-receptor subtypes D1, D2, D3 and the serotonergic-receptor subtypes 5-HT(1A), 5-HT(2A), and 5-HT(2C). The most active compounds showed MICs from 2 to 4 microg/mL and had overall reduced binding to the dopamine and serotonin receptors compared to chlorpromazine and trifluoperazine. PMID- 17407814 TI - Elongated patellae at the final stage of Osgood-Schlatter disease: a radiographic study. AB - Aetiology of Osgood-Schlatter disease (OSD) is still unknown. Relative length of the patellar articular surface with the length of the patella, relative height of the tibial tubercle with the sagittal diameter of the tibia, Insall-Salvati, Blackburne-Peel, and Caton-Deschamps indexes, as well as Grelsamer-morphology type of the patella were measured from preoperative plain X-rays in 82 knees of 20-year-old males with OSD and in 87 knees of 20-year-old male controls with normal MRI findings of the knee. Seventy-eight of the OSD patients had separate ossicles. Their mean patellar morphology index was 1.44 and that of the controls was 1.28 (p<0.001), indicating significant lengthening of the patellar body among the OSD group. OSD patients had also significantly more often Grelsamer type II (elongated patellae) than the controls. Tibial tuberosity was significantly higher among OSD patients, but was not correlated with the lengthening of the patella. An increased patellar height among OSD patients was shown by the Blackburne-Peel and Caton-Deschamps indexes (p<0.001) and the Insall-Salvati index (p=0.018). OSD patients exhibit elongated patellae and patellar tendons which may result from long-standing tension of the extensor apparatus during growth spurt, when femoral growth exceeds that of the anterior structures of the knee. PMID- 17407815 TI - Tricyclic isoxazolines: identification of R226161 as a potential new antidepressant that combines potent serotonin reuptake inhibition and alpha2 adrenoceptor antagonism. AB - In previous articles we have described the discovery of a new series of tricyclic isoxazolines combining central serotonin (5-HT) reuptake inhibition with alpha(2) adrenoceptor antagonistic activity. We report now on the synthesis, the in vitro binding potency and the primary in vivo activity of six enantiomers within this series, one of which was selected for further pharmacological evaluation and assigned as R226161. Some additional in vivo studies in rats are described with this compound, which proved to be centrally and orally active as a combined 5-HT reuptake inhibitor and alpha(2)-adrenoceptor antagonist. PMID- 17407816 TI - Synthesis and bioassay evaluation of 1-(4-substitutedideneaminooxymethyl)-phenyl 3-(2,6-difluorobenzoyl)ureas. AB - A variety of novel 1-(4-substitutedideneaminooxymethyl)-phenyl-3-(2,6 difluorobenzoyl)ureas were designed and synthesized by the reaction of 4 substitutedideneaminooxymethyl aniline with 2,6-difluorobenzoyl isocyanates in good yields. The title compounds were soluble in most organic solvents, which should make them easier to use. The preliminary bioassay showed that some of the title compounds show excellent insecticidal activity against Mythimna separata at the dosage of 25 mg kg(-1) and moderate insecticidal activity against Nephotettix cincticeps at the dosage of 500 mg kg(-1). Toxicity assays indicated that these title compounds cause in M. separata and N. cincticeps such symptoms of toxicity as discolouration, and weight loss, and cessation of feeding and lethal. One title compound exhibited acaricidal activity against Tetranychus urticae. PMID- 17407817 TI - Novel series of substituted biphenylmethyl urea derivatives as MCH-R1 antagonists for the treatment of obesity. AB - We have designed and synthesized two novel series of MCH-R1 antagonists based on a substituted biphenylmethyl urea core. SAR was explored, suggesting that optimal binding with the receptor was achieved when the biphenylmethyl group and the linker were substituted on the same nitrogen of the urea moiety. Compound 1-(3' cyano-4-biphenylmethyl)-3-(2-hydroxy-1,1-dimethylethyl)-1-{2-[1-(4-methylbenzyl) 4-piperidinyl]ethyl}urea 2t showed the best antagonist binding activity to the MCH-R1 with a 43 nM K(i). PMID- 17407818 TI - An exogenous marker: a novel approach for the characterization of oxidative stress. AB - Oxidative stress (OS) and its consequences which promote alterations in biomolecules, to tissue damage and to the development of pathological conditions, continue to attract many investigators. The identification of reliable biomarker is essential for the characterization of OS and possibly for early discovery of OS-associated diseases. The aim of the present study was to offer a new concept in the development of novel probes for OS, based on the design, synthesis, and utilization of exogenous markers, as alternative to the search for endogenous markers. This article describes: (a) the synthesis of such a marker, linoleoyl tyrosine 2-deoxyguanosyl ester (LTG), constructed from three endogenous subunits: linoleic acid, tyrosine, and 2'-deoxyguanosine, representing the three major groups from which the body is composed, unsaturated fatty acids (USFA), proteins, and DNA, respectively, all bound covalently and (b) the development of analytical tools (LC/MS/MS) to enable the identification of the different LTG oxidized products formed under OS by exposure of LTG to different reactive oxygen species (ROS) such as, copper ions and hypochlorous acid. PMID- 17407819 TI - A double catgrip mixed L and D mini protein only 20 residues long. AB - Stereochemistry limits but also defines proteins, as conformational constructs stereospecific for poly-L structure. Employed as a variable in sequence, stereochemistry could make proteins customizable in the letters of L and D amino acid alphabet. In proof of concept, we previously demonstrated stereochemical reengineering of canonical beta-hairpins as bracelet and boat shaped molecules. Illustrating the prospect for functional design, a 20-residue four-stranded mini beta protein is now customized stereochemically as a canoe shaped molecule. A conformational construct of four side by side hydrogen-bonded strands in alternately (l)beta, (d)beta conformation, joined via Type-II/II' beta-turns, is planned to be preponderantly apolar in beta-sheet favoring residues, interspersing two ion pairs, and suitably L and D in sequence. Synthesis followed by MD, NMR, CD, and MALDI-MS studies established the molecule as a canoe shaped fold in water, demonstrable in affinity of alkali and alkaline-earth metal ions as expected given its catgrip like elements. Another success in accomplishing a synthetic miniprotein complex in stereochemistry and stereospecific in conformation, exceptionally small yet functional in metal ion affinity, affirms the value in combined L and D alphabet in programming molecular shapes and functions stereochemically. PMID- 17407820 TI - Effect of low-level helium-neon laser therapy on histological and ultrastructural features of immobilized rabbit articular cartilage. AB - The present study investigates whether low-level helium-neon laser therapy can increase histological parameters of immobilized articular cartilage in rabbits or not. Twenty five rabbits were divided into three groups: the experiment group, which received low-level helium-neon laser therapy with 13J/cm(2) three times a week after immobilization of their right knees; the control group which did not receive laser therapy after immobilization of their knees; and the normal group which received neither immobilization nor laser therapy. Histological and electron microscopic examinations were performed at 4 and 7 weeks after immobilization. Depth of the chondrocyte filopodia in four-week immobilized experiment group, and depth of articular cartilage in seven-week immobilized experiment group were significantly higher than those of relevant control groups (exact Fisher test, p=0.001; student's t-test, p=0.031, respectively). The surfaces of articular cartilages of the experiment group were relatively smooth, while those of the control group were unsmooth. It is therefore concluded that low-level helium-neon laser therapy had significantly increased the depth of the chondrocyte filopodia in four-week immobilized femoral articular cartilage and the depth of articular cartilage in seven-week immobilized knee in comparison with control immobilized articular cartilage. PMID- 17407821 TI - Gene expression and characterization of 2-keto-3-deoxygluconate kinase, a key enzyme in the modified Entner-Doudoroff pathway of the aerobic and acidophilic hyperthermophile Sulfolobus tokodaii. AB - 2-Keto-3-deoxygluconate kinase (KDGK) catalyzes the ATP-dependent phosphorylation of 2-keto-3-deoxygluconate, a key intermediate in the modified (semi phosphorylative) Entner-Doudoroff (ED) glucose metabolic pathway. We identified the gene (ORF ID: ST2478) encoding KDGK in the hyperthermophilic archaeon Sulfolobus tokodaii based on the structure of a gene cluster in a genomic database and functionally expressed it in Escherichia coli. The expressed protein was purified from crude extract by heat treatment and two conventional column chromatography steps, and the partial amino acid sequence in the N-terminal region of the purified enzyme (MAKLIT) was identical to that obtained from the gene sequence. The purified enzyme was extremely thermostable and retained full activity after heating at 80 degrees C for 1 h. The enzyme utilized ATP or GTP, but not ADP or AMP, as a phosphoryl donor and 2-keto-3-deoxy-D-gluconate or 2 keto-D-gluconate as a phosphoryl acceptor. Divalent cations including Mg(2+), Co(2+), Ni(2+), Zn(2+) or Mn(2+) were required for activity, and the apparent Km values for KDG and ATP at 50 degrees C were 0.027 mM and 0.057 mM, respectively. The presence of KDGK means that the hyperthermophilic archaeon S. tokodaii metabolizes glucose via both modified (semi-phosphorylative) and non phosphorylative ED pathways. PMID- 17407822 TI - High-risk behaviors in women who use crack: knowledge of HIV serostatus and risk behavior. AB - PURPOSE: To determine whether crack cocaine-using women who are aware of their HIV serostatus have made modifications in risk behaviors, we compared known HIV positive (HIV+) and HIV-negative (HIV-) users with respect to sexual risk behaviors, prevalence of sexually transmitted infections (STIs) and vaginitis, and correlates of unprotected sex. METHODS: We used a cross-sectional design with street outreach, recruitment, and interviews of sexually active crack cocaine using women. Women received testing for HIV, STIs, and vaginitis. RESULTS: Sixty one HIV+ and 117 HIV- women were enrolled. HIV+ women were significantly more likely to be African-American. There were no significant differences in drug use, types of sexual partners, number of paying partners, attitudes regarding condoms, or STI diagnoses. HIV+ women were less likely to engage in unprotected sex compared with HIV- women (56% vs. 75%, adjusted odds ratio [AOR], 0.36; 95% confidence interval [CI], 0.13-0.99). Among HIV+ women, unprotected sex was negatively associated with stronger beliefs regarding the protective value of condoms (AOR, 0.07; 95% CI, 0.01-0.67) and concurrent injection-drug use (AOR, 0.19; 95% CI, 0.04-0.99). CONCLUSIONS: Although the majority of crack using HIV+ crack using women in this sample continued to engage in high-risk sexual activities, they were less likely to do so than HIV- women. Interventions targeting this population are needed. PMID- 17407823 TI - Presentation duration and false recall for semantic and phonological associates. AB - Two experiments examined false recall for lists of semantically and phonologically associated words as a function of presentation duration. Veridical recall increased with long exposure durations for all lists. For semantically associated lists, false recall increased from 20-250 ms, then decreased. There was a high level of false recall with 20 ms durations for phonologically associated lists (47 and 44% for Experiments 1 and 2, respectively), which declined as duration increased. In Experiment 2, for lists presented at 20 and 50 ms rates, false recall given zero correct recall was observed frequently, suggesting that conscious recollection of studied words was not necessary for phonological false memory. Differences between phonologically and semantically associated lists were consistent with a feature integration model based on automatic initial processing of phonetic features of words. PMID- 17407824 TI - The neural correlates of sign versus word production. AB - The production of sign language involves two large articulators (the hands) moving through space and contacting the body. In contrast, speech production requires small movements of the tongue and vocal tract with no observable spatial contrasts. Nonetheless, both language types exhibit a sublexical layer of structure with similar properties (e.g., segments, syllables, feature hierarchies). To investigate which neural areas are involved in modality independent language production and which are tied specifically to the input output mechanisms of signed and spoken language, we reanalyzed PET data collected from 29 deaf signers and 64 hearing speakers who participated in a series of separate studies. Participants were asked to overtly name concrete objects from distinct semantic categories in either American Sign Language (ASL) or in English. The baseline task required participants to judge the orientation of unknown faces (overtly responding 'yes'/'no' for upright/inverted). A random effects analysis revealed that left mesial temporal cortex and the left inferior frontal gyrus were equally involved in both speech and sign production, suggesting a modality-independent role for these regions in lexical access. Within the left parietal lobe, two regions were more active for sign than for speech: the supramarginal gyrus (peak coordinates: -60, -35, +27) and the superior parietal lobule (peak coordinates: -26, -51, +54). Activation in these regions may be linked to modality-specific output parameters of sign language. Specifically, activation within left SMG may reflect aspects of phonological processing in ASL (e.g., selection of hand configuration and place of articulation features), whereas activation within SPL may reflect proprioceptive monitoring of motoric output. PMID- 17407825 TI - Capsaicin-induced thermal hyperalgesia and sensitization in the human trigeminal nociceptive pathway: an fMRI study. AB - The aim of this study was to differentiate the processing of nociceptive information, matched for pain intensity, from capsaicin-induced hyperalgesic vs. control skin at multiple levels in the trigeminal nociceptive pathway. Using an event-related fMRI approach, 12 male subjects underwent three functional scans beginning 1 h after topical application of capsaicin to a defined location on the maxillary skin, when pain from capsaicin application had completely subsided. Brush and two levels of painful heat (low-Thermal-1 and high-Thermal-2) were applied to the site of capsaicin application and to the mirror image region on the opposite side. Temperatures for each side were set to evoke perceptually matched pain (mean temperatures [capsaicin/control]: Thermal-1=38.4/42.8 degrees C; Thermal-2=44.9/47.8 degrees C). We found differences in activation patterns following stimuli to treated and untreated sides in sensory circuits across all stimulus conditions. Across the trigeminal nociceptive pathway, Thermal-2 stimulation of hyperalgesic skin evoked greater activation in trigeminal ganglion and nucleus, thalamus, and somatosensory cortex than the control side. Thus, trigeminal nociceptive regions showed increased activation in the context of perceptually equal pain levels. Beyond these regions, contrast analyses of capsaicin vs. control skin stimulation indicated significant changes in bilateral dorsolateral prefrontal cortex and amygdala. The involvement of these emotion related regions suggests that they may be highly sensitive to context, such as prior experience (application of capsaicin) and the specific pain mechanism (hyperalgesic vs. normal skin). PMID- 17407826 TI - Totally laparoscopic aortohepatic bypass for aortic debranching during endovascular thoracoabdominal aneurysm repair. AB - INTRODUCTION: Endovascular grafting of the aorta is gaining widespread acceptance for treating aortic aneurysms. Para-renal aneurysms or thoraco-abdominal aneurysms may be a relative contra-indication for endovascular aneurysm repair (EVAR) unless visceral vessels can be debranched. REPORT: We describe a case of thoraco-abdominal aneurysm extending from the descending thoracic aorta to the level of coeliac artery. A totally laparoscopic retrograde aorto-hepatic bypass was performed in conjunction with endograft exclusion of the aneurysm and coverage of the coeliac artery ostium. DISCUSSION: Laparoscopic debranching of visceral vessels extends the indications of EVAR. PMID- 17407827 TI - Rupture of abdominal aortic aneurysms. What matters most: geometry or blood pressure? PMID- 17407828 TI - Energy and distribution in subterranean rodents: sympatry between two species of the genus Ctenomys. AB - The low basal metabolic rate (BMR) observed in subterranean rodents, compared to that of surface-dwelling species of comparable size, has been proposed to be an adaptation to underground life. Two main hypotheses have been proposed to explain this finding, the cost of burrowing and the thermal stress. The former states that the low BMR is due to the high cost of extending the tunnel system whereas the other relates it to the possibility of overheating in burrows where evaporative and convective heat exchange are restricted. Additionally, both hypotheses related the energetics of subterranean rodent with spatial distribution. The genus Ctenomys is an excellent model to evaluate the cost of burrowing or thermal stress, since they are widely distributed, with members differing markedly in body mass. The aim of this study was to assess digging and basal energetics in two Ctenomys species that live in sympatry in a coastal grassland, but differ in their microspatial distribution by soil preference. We used the obtained energetic data to test both energy-distribution hypotheses. We measured BMR and digging metabolic rate (DMR) through open flow respirometry in two species exposed to soft and hard soils. In brief, DMR in Ctenomys talarum (100-170 g), as in Ctenomys australis (250-600 g), was unaffected by soil hardness. Within thermoneutral zone of each species, DMR/RMR quotient was lower in the smaller species. Our data did not support the thermal stress hypothesis, but the cost of burrowing hypothesis was not rejected. Other alternative hypotheses are proposed to explain the distribution of C. talarum and C. australis. PMID- 17407829 TI - The response of digestive proteases to abrupt salinity decrease in the euryhaline sparid Sparus aurata L. AB - The response of the digestive proteases to abrupt salinity change was studied in juvenile gilthead sea bream (Sparus aurata) for 15 days after transfer from 33 per thousand to 21 per thousand. Salinity decrease affected significantly neither the activity of total acid proteases in stomach, nor the activities of total alkaline proteases and major serine proteases--trypsin and chymotrypsin--in the alkaline part of the intestine. The activity of the major proteases was significantly different between the alkaline segments of the intestine, with the posterior intestine presenting the highest activities followed by the pyloric caeca. This distribution pattern remained unaffected by salinity decrease. Notably, salinity change led to significant alterations in elastase and carboxypeptidase activity. The changes were more prominent in the upper part of the intestine (pyloric caeca and anterior intestine) than in the posterior intestine. In pyloric caeca significant alteration of carboxypeptidase A and B activities was observed, elastase changes were confined to anterior intestine together with alterations in carboxypeptidase B activity, while in posterior intestine the changes were restricted to carboxypeptidase A activity. The results are discussed in relation to the osmoregulatory action of the intestinal segments and dietary protein digestion. PMID- 17407830 TI - Cardiomyocyte-restricted over-expression of C-type natriuretic peptide prevents cardiac hypertrophy induced by myocardial infarction in mice. AB - OBJECTIVE: Infused C-type natriuretic peptide (CNP) was recently found to play a cardioprotective role in preventing myocardial ischaemia/reperfusion (I/R) injury and improving cardiac remodelling after myocardial infarction (MI) in rats. Our study aimed to investigate the effect of cardiomyocyte-specific CNP over expression on I/R injury and MI in transgenic mice. METHODS AND RESULTS: We generated transgenic (TG) mice over-expressing CNP in cardiomyocytes. Elevated CNP expression on RNA and protein levels was demonstrated by RNase-protection assay and radioimmunoassay. Male TG mice and age-matched wild-type (WT) littermates were subjected to 1-hour global myocardial ischaemia and 23 h of reperfusion or permanent ligation of the coronary artery for 3 weeks. Infarct size did not differ between the WT and TG groups in mice subjected to I/R. In mice that underwent permanent ligation of coronary arteries, both left and right ventricular hypertrophy were prevented by CNP over-expression 3 weeks post-MI. Histological analysis revealed less necrosis, muscular degeneration and inflammation in infarcted TG mice. Impairment of cardiac function was less pronounced in transgenic animals than in the wild-type controls. CONCLUSIONS: Over-expression of CNP in cardiomyocytes does not affect I/R-induced infarct size but prevents cardiac hypertrophy induced by MI. Therefore, CNP may represent a potent therapeutic target for the treatment of patients with cardiac hypertrophy induced by myocardial infarction or other aetiology. PMID- 17407831 TI - Sustiva: revised drug-interaction information. AB - The FDA added new drug-interaction warnings to the prescribing information for Sustiva (efavirenz). PMID- 17407832 TI - Prisoner death rate very high immediately after release. AB - During the first two weeks after release, prisoners in Washington State had 129 times the death rate from drug overdose, compared to other state residents- probably because they did not know how much less drug they could tolerate, after taking little or none of it in prison. Cardiovascular disease, homicide, suicide, cancer, and traffic accidents also caused excessive deaths. PMID- 17407833 TI - CTS - Israel and Ireland, encountering the same problem. PMID- 17407834 TI - Epithelial responses to oxidative stress in chronic obstructive pulmonary disease: lessons from expression profiling. PMID- 17407835 TI - Extending the good diet, good health paradigm: modulation of breast cancer resistance protein (BCRP) by flavonoids. PMID- 17407836 TI - [Comment on drainage insertion study in otitis media]. PMID- 17407837 TI - [Cancer in Denmark]. PMID- 17407838 TI - Comprehensive diagnosis of Rett's syndrome relying on genetic, epigenetic and expression evidence of deficiency of the methyl-CpG-binding protein 2 gene: study of a cohort of Israeli patients. AB - BACKGROUND: Despite advances in the characterisation of mutations in the MECP2 coding region, a small proportion of classic RTT cases remain without recognisable mutations. OBJECTIVE AND METHODS: To identify previously unknown mutations, a quantitative assay was established, providing estimates of MECP2_e1 and MECP2_e2 expression levels in peripheral blood. A systematic analysis of an Israeli cohort of 82 patients with classic and atypical RTT is presented, including sequence analysis of the MECP2-coding region, MLPA, XCI and quantitative expression assays. RESULTS AND CONCLUSION: A novel mis-sense mutation at ca 453C-->T (pD151E), resulting in a change of a conserved residue at the methyl-binding domain, and a rare GT deletion of intron 1 donor splice site are reported. It is shown that various MECP2 mutations had distinct effects on MECP2 expression levels in peripheral blood. The most significant (p<0.001) reduction in the expression of both MECP2 isoforms was related to the presence of the intron 1 donor splice-site mutation. Using quantitative expression assays, it was shown that several patients with classic and atypical RTT with no mutation findings had significantly lower MECP2 expression levels. Further research on these patients may disclose still elusive non-coding regulatory MECP2 mutations. PMID- 17407839 TI - Systematic experimental based modeling of a rotary piezoelectric ultrasonic motor. AB - In this paper, a new method for equivalent circuit modeling of a traveling wave ultrasonic motor is presented. The free stator of the motor is modeled by an equivalent circuit containing complex circuit elements. A systematic approach for identifying the elements of the equivalent circuit is suggested. The Levenberg Marquardt parameter estimation algorithm is used to model the alteration of the admittance after placing the rotor on the stator. Thereafter, theoretical assessments and experimental measurements are used to account for the speed reduction that is caused by placing the rotor on the stator and applying the load torque. Finally, the effects of temperature changes and the resultant response of the motor are computed. Results of the experiments and measurements are used to verify and validate the precision of the new modeling method. PMID- 17407840 TI - A new mechanism design of electro-magnetic actuator for a micro-positioner. AB - This paper proposes a new type of non-conventional electro-magnetic actuator (EMA) for micro-positioning. It is a repulsive magnetic system consisting of a motion pad and two active coils. The motion of the pad is caused by the repelling force between the coils and the permanent magnets on the pad. First, the dynamic model is derived and analyzed. Next, an adaptive sliding mode controller is developed to deal with the unknown parameters with the objective of precision positioning. The experimental results demonstrate satisfactory performances including regulation accuracy and control stiffness for the system. PMID- 17407841 TI - Solution of nonlinear optimal control problems using a semi-exhaustive search. AB - The solution of nonlinear optimal control problems is a challenging problem. The recommended practice is to use different methods and then to implement the best solution found. To support this objective, the performance of a semi-exhaustive search method that uses a different approach is presented in this paper. The performance is compared against an Iterative Dynamic Programming (IDP) algorithm on five example problems. The semi-exhaustive search provided a smooth convergence to the optimal solution and required a significantly reduced computational time as compared with the IDP algorithm. PMID- 17407845 TI - Re: a "Perfect storm". PMID- 17407846 TI - "Whipple's disease". PMID- 17407847 TI - Prices of generic intraocular lenses likely to see gradual decline. PMID- 17407848 TI - Is your advertising a liability risk? PMID- 17407849 TI - Dr. Albert Schweitzer: moral guardian. PMID- 17407850 TI - Pollen to deliver drugs. PMID- 17407851 TI - Let's stop talking - our patients are waiting. PMID- 17407852 TI - Outbreak news. Rift Valley fever, United Republic of Tanzania. PMID- 17407853 TI - Elimination of measles in the Republic of Korea, 2001-2006. PMID- 17407854 TI - It's tough to make predictions... PMID- 17407855 TI - Statin wars following coronary revascularization - evidence-based clinical practice? PMID- 17407856 TI - [Comment on Butterwegge: Continuous monitoring of status sub partu--discontinuing fetal pulse oximetry? Z Geburtshilfe Neonatol 2006; 210: 119-120. ]. PMID- 17407857 TI - Consumer tolerance for inaccuracy in physician performance ratings: one size fits none. AB - Health plans increasingly use physician performance ratings, but some physicians are concerned that measurement inaccuracies may jeopardize their reputations and livelihoods. Absent from the debate thus far are consumer views about how accurate physician ratings need to be for various uses. Consumer tolerance for inaccuracy in physician performance ratings varies widely, according to a new national study by the Center for Studying Health System Change (HSC). At least one-third of adults have a low tolerance for inaccuracy (5 percent or less), but more than one of every five adults would tolerate ratings that were 20 percent-50 percent inaccurate. Consumers' relatively higher tolerance for inaccuracy when used for public reporting and tiered networks may speed these uses of physician performance ratings by health plans. However, consumers' lower tolerance for inaccurate ratings when choosing their own physicians and paying physicians for performance may hinder such uses. PMID- 17407858 TI - A passion for public health. PMID- 17407859 TI - North Carolina shares a model for dealing with disaster. PMID- 17407860 TI - Fire departments add pet oxygen masks to equipment checklists. PMID- 17407861 TI - Metabolic syndrome and CRP. PMID- 17407863 TI - Comment on "The directionality of acoustic T-phase signals from small magnitude submarine earthquakes" [J. Acoust. Soc. Am. 119, 3669-3675 (2006)]. AB - In a recent paper, Chapman and Marrett [J. Acoust. Soc. Am. 119, 3669-3675 (2006)] examined the tertiary (T-) waves associated with three subduction-related earthquakes within the South Fiji Basin. In that paper it is argued that acoustic energy is radiated into the sound channel by downslope propagation along abyssal seamounts and ridges that lie distant to the epicenter. A reexamination of the travel-time constraints indicates that this interpretation is not well supported. Rather, the propagation model that is described would require the high-amplitude T-wave components to be sourced well to the east of the region identified, along a relatively flat-lying seafloor. PMID- 17407864 TI - A pseudo-inverse algorithm for simultaneous measurements using multiple acoustical sources. AB - Simultaneous multiple acoustical sources measurement (SMASM) has been proposed for more effective and reliable identification of acoustical systems under critical conditions [N. Xiang and M. R. Schroeder, J. Acoust. Soc. Am. 113, 2754 2761 (2003); N. Xiang, J. N. Daigle, and M. Kleiner, J. Acoust. Soc. Am. 117, 1889-1894 (2005)]. This paper presents a pseudo-inverse algorithm for the SMASM correlation technique as an alternative way of extracting impulse responses of acoustical channels. Simulations and room acoustics experiments are carried out and the results prove the feasibility of the proposed algorithm. PMID- 17407865 TI - Characteristics of penalty mode scattering by rigid splices in lined ducts. AB - In lined ducts, incident modes are scattered by axially and circumferentially nonuniform impedance. Experiments and numerical calculations have proved that this mode scattering can reduce the liner performance in some cases. This paper is devoted to the characterization of the penalty mode scattering excited by hard walled splices which often exist in lined ducts. It is shown that, in the range of small splice angles, the transmission loss may decrease sharply with increasing splice angle when one mode, which is near cut-off or has high azimuthal order, is incident. When the incident sound field is composed of several acoustical modes, the phase interferences of incident modes are important for the penalty mode scattering. The effects of other parameters, e.g., liner length, mode quasiresonance on the penalty mode scattering are also presented. PMID- 17407866 TI - Finite element analysis of guided waves in fluid-filled corrugated pipes. AB - Free wave propagation in fluid-filled corrugated pipes is analyzed using finite element methods in combination with a wave-based approach. By combining discretized models with a wave-based approach, complex mechanism of wave motion in the three-dimensional waveguide is fully included. The pipes are treated as waveguides having periodic properties in the direction of wave propagation. The analysis of these guided waves leads to dispersion curves which show the strong frequency-dependency of the different wave modes. The method also allows the inclusion of coupling between fluid-borne and structure-borne wave modes which occur at the acoustic-structure interface. Phase and group velocities of the wave modes are derived in postprocessing steps. Additionally, the energy ratio of the fluid-domain and solid-domain vibrational energies is computed. Finally, linear damping models are included in order to explore wave mode attenuation. PMID- 17407867 TI - Radiation force and shear motions in inhomogeneous media. AB - An action of radiation force induced by ultrasonic beam in waterlike media such as biological tissues (where the shear modulus is small as compared to the bulk compressibility) is considered. A new, nondissipative mechanism of generation of shear displacement due to a smooth (nonreflecting) medium inhomogeneity is suggested, and the corresponding medium displacement is evaluated. It is shown that a linear primary acoustic field in nondissipative, isotropic elastic medium cannot excite a nonpotential radiation force and, hence, a shear motion, whereas even smooth inhomogeneity makes this effect possible. An example is considered showing that the generated displacement pulse can be significantly longer than the primary ultrasound pulse. It is noted that, unlike the dissipative effect, the nondissipative action on a localized inhomogeneity (such as a lesion in a tissue) changes its sign along the beam axis, thus stretching or compressing the focus area. PMID- 17407868 TI - Noncontact transportation in water using ultrasonic traveling waves. AB - A noncontact transport experiment in water using ultrasonic traveling waves was investigated. Acrylic, aluminum, and brass discs were used as test objects. Traveling waves were generated using two ultrasonic transducers attached at the ends of a vibrating plate. One side was used as the wave-source side and the other side was used as the wave-receiving side. Acrylic plates cemented to the sides of the vibrating plate formed a tank to hold water. Object transportation was accomplished by adding a small amount of water to the vibrating structure. The transport velocity of floating objects in water is faster than for floating transport in air because of buoyancy. The transport velocity of an object depends on water height. The minimum value of the velocity occurs when the disc thickness is equal to the water height. The transport velocity increases as the height of water increases. For very shallow depths, the largest velocity is obtained when cavitation-induced streaming occurs. PMID- 17407869 TI - Experiments on sound generation in corrugated pipes with flow. AB - The article reports acoustic measurements on short corrugated pipes with flow. Such pipes might generate high sound levels associated with length resonances. One of the main objectives of the study was to estimate the location of the effective sources by studying the energy flow through the pipes. It was found that a short section of corrugations will only produce sound effectively when placed at the inflow end, while for fully corrugated pipes, the sound-producing regions are located around the pressure maxima of the observed standing waves. It was further found that the net energy flow is in the upstream direction for nearly the complete length of pipe. PMID- 17407870 TI - Vortex sound due to a flexible boundary backed by a cavity in a low mach number mean flow. AB - Low frequency sound radiated due to the unsteady motion of an inviscid vortex in the proximity of a flexible membrane backed by an airtight cavity on an otherwise rigid plane is investigated theoretically. Results show that both monopole and dipole are created but the latter is important only when the vortex is traversing over the membrane. The monopole results from the membrane vibration and the dipole from the transverse motion of the vortex. It is also found that these sound fields tend to counteract each other. The increase in the mean flow speed in general results in a stronger acoustic power radiation, but sound attenuation may be possible if the membrane-cavity system is weak compared with the mean flow momentum. PMID- 17407871 TI - A computational method of evaluating noncompact sound based on vortex sound theory. AB - A numerical investigation is made of the production of sound by turbulence interacting with a noncompact body. The problem is formulated in the frequency domain by extending the theory of vortex sound proposed by Howe. The anomalous "numerical" generation of sound by the sudden termination of Lighthill's stress tensor at the outer boundary of a finite computational domain is avoided by identification of "scattered" sound sources that generate sound principally by interaction with the solid surface. It is argued that the boundary element method is the most efficient means of computing the aeroacoustic Green's function for the problem, because it requires a minimum of CPU time, is not prone to numerical errors such as dispersion and dissipation during propagation, and the radiation condition is easily applied at the outer boundary. The method is applied to the problem of sound generation by high Reynolds number flow past a circular cylinder. The "scattered" sources are shown to be confined to the vicinity of the cylinder surface. At low frequencies the radiation has a dipole-like directivity in agreement with the compact approximation. However, the directivity is quite different at high frequencies, where our noncompact method predicts a more complicated "leaf-like" radiation pattern. PMID- 17407872 TI - Multipath pulse shapes in shallow water: theory and simulation. AB - In shallow water propagation the steeper ray angles are weakened most by boundary losses. Regarding the sound intensity as a continuous function of angle it can be converted into a function of travel time to reveal the multipath pulse shape received from a remote source (one-way path) or a target (two-way path). The closed-form isovelocity pulse shape is extended here to the case of upward or downward refraction. The envelope of the earliest arrivals is roughly trapezoidal with a delayed peak corresponding to the slowest, near horizontal refracted paths. The tail of the pulse falls off exponentially (linearly in decibels) with a decay constant that depends only on the bottom reflection properties and water depth, irrespective of travel time, a useful property for geoacoustic inversion and for sonar design. The nontrivial analytical problem of inverting explicit functions of angle into explicit functions of time is solved by numerical interpolation. Thus exact solutions can be calculated numerically. Explicit closed-form approximations are given for one-way paths. Two-way paths are calculated by numerical convolution. Using the wave model C-SNAP in several broadband cases of interest it is demonstrated that these solutions correspond roughly to a depth average of multipath arrivals. PMID- 17407873 TI - Propagation in an elastic wedge using the virtual source technique. AB - The virtual source technique, which is based on the boundary integral method, provides the means to impose boundary conditions on arbitrarily shaped boundaries by replacing them by a collection of sources whose amplitudes are determined from the boundary conditions. In this paper the virtual source technique is used to model propagation of waves in a range-dependent ocean overlying an elastic bottom with arbitrarily shaped ocean-bottom interface. The method is applied to propagation in an elastic Pekeris waveguide, an acoustic wedge, and an elastic wedge. In the case of propagation in an elastic Pekeris waveguide, the results agree very well with those obtained from the wavenumber integral technique, as they do with the solution of the parabolic equation (PE) technique in the case of propagation in an acoustic wedge. The results for propagation in an elastic wedge qualitatively agree with those obtained from an elastic PE solution. PMID- 17407874 TI - Gray whale target strength measurements and the analysis of the backscattered response. AB - One of the current Integrated Marine Mammal Monitoring and Protection System (IMPAS) directions is concentrated on the design and development of the active sonar modality representing just one component of the global system. The active sonar was designed, built, and tested during the Marine Mammal Active Sonar Test (MAST 04), producing whale detections and whale tracks. The experiment was conducted in January 2004 off the coast of California. One of the objectives of the current work is to distinguish whale backscattered responses from the ones generated by the environmental clutter in a waveguide. Furthermore, the work aims to identify and analyze the target signature features that are necessary for enhanced active sonar detection and classification of marine mammals. Over the years there have been very few documented attempts to capture and analyze the backscattering response of whales using an active sonar system. Nevertheless, whales, mostly owing to their size, their motion, and the aspect dependence of their backscattered field, possess desirable properties that help distinguish their scattered response from clutter and other environment related false alarms. As an initial step, data collected during the MAST 04 experiment are presented, and gray whale target strength measurements are obtained. Results are compared to the previously published whale target strengths. Additionally, an investigation is conducted in an effort to provide whale feature identification points suitable for automated detection and classification, as means of relating gray whale active acoustic signatures to their inherent characteristics and their motion. PMID- 17407875 TI - Miniaturized mandrel-based fiber optic hydrophone. AB - Fiber optic hydrophones typically use a long sensor fiber wrapped around an air filled mandrel to achieve high responsivity. Minimum mandrel diameter is determined by fiber reliability considerations. A miniature hydrophone, with diameter of about a third that of previous hydrophones, is reported. Such a hydrophone has applications for very thin towed arrays. Small diameter fiber with high strength Bragg gratings written through the coating are key enablers. PMID- 17407876 TI - Aberrations in materials with random inhomogeneities. AB - Materials that consist of a random microstructure can affect ultrasonic measurements--reducing signal strength, increasing noise, and reducing measurement accuracy--through scattering and aberration of the acoustic field. To account for these adverse effects a phase screen model, alongside the stochastic wave equation, has been developed. This approach allows the field and study aberrations to be modeled from a statistical point of view. Experimental evidence of aberration and statistical properties of the measured acoustic field are shown. A measured correlation function of the acoustic field is interlinked to mean crystallite size by using a theoretical coherence function that can be mainly described by the correlation length and wave velocity variation of microstructure. The estimation of the mean crystallite size using this technique would provide some insight into material characterization. PMID- 17407877 TI - Reproducible cavitation activity in water-particle suspensions. AB - The study of cavitation inception in liquids rarely yields reproducible data, unless special control is taken on the cleanliness of the experimental environment. In this paper, an experimental technique is demonstrated which allows repeatable measurements of cavitation activity in liquid-particle suspensions. In addition, the method is noninvasive: cavitation bubbles are generated using a shock-wave generator, and they are photographed using a digital camera. The cavitation activity is obtained after suitable image processing steps. From these measurements, the importance of the particle's surface structure and its chemical composition is revealed, with polystyrene and polyamide particles generating the highest yields. Further findings are that cavitation nuclei become depleted with an increasing number of experiments, and the existence of nuclei with varying negative pressure thresholds. Finally, a decrease of the cavitation yield is achieved by prepressurization of the suspension-indicating that the cavitation nuclei are gaseous. PMID- 17407878 TI - Parallel capillary-tube-based extension of thermoacoustic theory for random porous media. AB - Thermoacoustic theory is extended to stacks made of random bulk media. Characteristics of the porous stack such as the tortuosity and dynamic shape factors are introduced into the thermoacoustic wave equation in the low reduced frequency approximation. Basic thermoacoustic equations for a bulk porous medium are formulated analogously to the equations for a single pore. Use of different dynamic shape factors for the viscous and thermal effects is adopted and scaling using the dynamic shape factors and tortuosity is demonstrated. Comparisons of the calculated and experimentally derived thermoacoustic properties of reticulated vitreous carbon and aluminum foam show good agreement. A consistent mathematical model of sound propagation in a random porous medium with an imposed temperature is developed. This treatment leads to an expression for the coefficient of the temperature gradient in terms of scaled cylindrical thermoviscous functions. PMID- 17407879 TI - Near-field ultrasound tomography. AB - In this paper, a near-field tomographic solution is introduced to solve the imaging problem of fluid objects assumed to be weakly heterogeneous (Born approximation) and excited by spherical waves. The solution to the forward problem is based on the Huygens-Fresnel principle which describes the scattered field as the result of the interference scheme of all the secondary spherical waves. From the derivation of the scattered field, a new Fourier transform that has been called the elliptical Fourier transform is defined: It differs from the standard Fourier transform in that instead of a plane wave decomposition, a harmonic ellipsoidal wave decomposition is obtained. Based on this spectral analysis, a near-field Radon transform is designed that complements the "far field tools" published in diffraction tomography literature. Then, assuming that the measuring distance is greater than one wavelength, the feasibility of reconstructing either the impedance or the velocity maps of an acoustical (perfect fluid) model is demonstrated. Numerical simulations were performed which confirmed the validity of the theory presented here; a theory which has many potential applications in future wave theory research. PMID- 17407880 TI - Acoustic power calibration of high-intensity focused ultrasound transducers using a radiation force technique. AB - To address the challenges associated with measuring the ultrasonic power from high-intensity focused ultrasound transducers via radiation force, a technique based on pulsed measurements was developed and analyzed. Two focused ultrasound transducers were characterized in terms of an effective duty factor, which was then used to calculate the power during the pulse at high applied power levels. Two absorbing target designs were used, and both gave comparable results and displayed no damage and minimal temperature rise if placed near the transducer and away from the focus. The method yielded reproducible results up to the maximum pulse power generated of approximately 230 W, thus allowing the radiated power to be calibrated in terms of the peak-to-peak voltage applied to the transducer. PMID- 17407881 TI - Elastic waves excited by a plane source on the surface of a multilayered medium. AB - Elastic waves excited by a plane piezoelectric source with an arbitrary shape on the surface of a multilayered medium have been studied for the first time in this paper. On the basis of Abzo-zena [Geophys. J. R. Astron. Soc. 58, 91-105 (1979)] and Menke [Geophys. J. R. Astron. Soc. 59, 315-323 (1979)], the propagator matrix for the elastic wave field in multilayered medium is extended from two- to three dimensional (3D) space. 3D elastic wave field is investigated and the displacement-stress response for the boundary conditions is obtained. The propagation of elastic wave in multilayered media is analyzed in 3D space in the frequency domain. The P-SV and SH modes corresponding to the poles are studied. The excitation and propagation of the modes are analyzed further. It is found that the propagation velocities of the P-SV and SH modes do not depend on the propagation azimuth theta in the plane parallel to the free surface of the multilayered medium while the displacement amplitudes are strongly dependent on the azimuth theta. The directional distribution functions of the modes are independent of the medium parameters and the modes and dependent on the shape and excitation fashion of the source. Finally, as an example, the displacement fields of the P-SV and SH modes excited by a rectangle source are analyzed. The displacement representation and numerical results of the directivity distribution functions for the P-SV and SH modes are obtained. PMID- 17407882 TI - Simulation of guided waves in complex piping geometries using the elastodynamic finite integration technique. AB - Although many technologies exist for inspecting piping systems, they are most successful on straight pipes and are often unable to accommodate the added complexities of pipe elbows, bends, twists, and branches, particularly if the region of interest is inaccessible. This paper presents a numerical technique based on the elastodynamic finite integration technique for simulating guided elastic wave propagation in piping systems. Comparisons show agreement between experimental and simulated data, and guided wave interaction with flaws, focusing, and propagation in pipe bends are presented. These examples demonstrate the ability of the simulation method to be used to study elastic wave propagation in piping systems which include three-dimensional pipe bends, and suggest its potential as a design tool for designing pipe inspection hardware and ultrasonic signal processing algorithms. PMID- 17407883 TI - A moving zone of quiet for narrowband noise in a one-dimensional duct using virtual sensing. AB - A frequent problem in active noise control is that the zone of quiet created at the error sensor tends to be very small. This means that the error sensor generally needs to be located close to an observer's ear, which might not always be a convenient or feasible solution. Virtual sensing is a method that can move the zone of quiet away from the error sensor to a desired location that is spatially fixed. This method has been investigated previously, and has shown potential to improve the performance of an active noise control system. However, it is very likely that the desired location of the zone of quiet is not spatially fixed. An active noise control system incorporating a virtual sensing method thus has to be able to create a moving zone of quiet that tracks the observer's ears. This paper presents a method for creating a moving zone of quiet based on the LMS virtual microphone technique. To illustrate the proposed method, it is implemented in an acoustic duct and narrowband control results are presented. These results show that a moving zone of quiet was effectively created inside the duct for narrowband noise. PMID- 17407884 TI - Improved precision in measurements of acoustic impedance spectra using resonance free calibration loads and controlled error distribution. AB - Resonances and/or singularities during measurement and calibration often limit the precision of acoustic impedance spectra. This paper reviews and compares several established techniques, and describes a technique that incorporates three features that considerably improve precision. The first feature is to minimize problems due to resonances by calibrating the instrument using up to three different acoustic reference impedances that do not themselves exhibit resonances. The second involves using multiple pressure transducers to reduce the effects of measurement singularities. The third involves iteratively tailoring the spectrum of the stimulus signal to control the distribution of errors across the particular measured impedance spectrum. Examples are given of the performance of the technique on simple cylindrical waveguides. PMID- 17407885 TI - Calibration sphere for low-frequency parametric sonars. AB - The problem of calibrating parametric sonar systems at low difference frequencies used in backscattering applications is addressed. A particular parametric sonar is considered: the Simrad TOPAS PS18 Parametric Sub-bottom Profiler. This generates difference-frequency signals in the band 0.5-6 kHz. A standard target is specified according to optimization conditions based on maximizing the target strength consistent with the target strength being independent of orientation and the target being physically manageable. The second condition is expressed as the target having an immersion weight less than 200 N. The result is a 280-mm-diam sphere of aluminum. Its target strength varies from -43.4 dB at 0.5 kHz to -20.2 dB at 6 kHz. Maximum excursions in target strength over the frequency band due to uncertainty in material properties of the sphere are of order +/-0.1 dB. Maximum excursions in target strength due to variations in mass density and sound speed of the immersion medium are larger, but can be eliminated by attention to the hydrographic conditions. The results are also applicable to the standard-target calibration of conventional sonars operating at low-kilohertz frequencies. PMID- 17407886 TI - Imaging with unfocused regions of focused ultrasound beams. AB - This article gives an analytical, computational, and experimental treatment of the spatial resolution encoded in unfocused regions of focused ultrasound beams. This topic is important in diagnostic ultrasound since ultrasound array systems are limited to a single transmit focal point per acoustic transmission, hence there is a loss of spatial resolution away from the transmit focus, even with the use of dynamic receive focusing. It is demonstrated that the spatial bandwidth of a Gaussian-apodized beam is approximately constant with depth, which means that there is just as much encoded spatial resolution away from the transmit focus as there is in the focal region. The practical application of this principle is discussed, an algorithm for retrospectively focusing signals from unfocused regions of fixed-focus beams is presented, and a quantitative comparison between the authors' methods and dynamic-receive beamforming is provided. PMID- 17407887 TI - Spectrum analysis of seismic surface waves and its applications in seismic landmine detection. AB - In geophysics, spectrum analysis of surface waves (SASW) refers to a noninvasive method for soil characterization. However, the term spectrum analysis can be used in a wider sense to mean a method for determining and identifying various modes of seismic surface waves and their properties such as velocity, polarization, etc. Surface waves travel along the free boundary of a medium and can be easily detected with a transducer placed on the free surface of the boundary. A new method based on vector processing of space-time data obtained from an array of triaxial sensors is proposed to produce high-resolution, multimodal spectra from surface waves. Then individual modes can be identified in the spectrum and reconstructed in the space-time domain; also, reflected waves can be separated easily from forward waves in the spectrum domain. This new SASW method can be used for detecting and locating landmines by analyzing the reflected waves for resonance. Processing examples are presented for numerically generated data, experimental data collected in a laboratory setting, and field data. PMID- 17407888 TI - Separation of components from impulses in reassigned spectrograms. AB - Two computational methods for pruning a reassigned spectrogram to show only quasisinusoidal components, or only impulses, or both, are presented mathematically and provided with step-by-step algorithms. Both methods compute the second-order mixed partial derivative of the short-time Fourier transform phase, and rely on the conditions that components and impulses are each well represented by reassigned spectrographic points possessing particular values of this derivative. This use of the mixed second-order derivative was introduced by Nelson [J. Acoust. Soc. Am. 110, 2575-2592 (2001)] but here our goals are to completely describe the computation of this derivative in a way that highlights the relations to the two most influential methods of computing a reassigned spectrogram, and also to demonstrate the utility of this technique for plotting spectrograms showing line components or impulses while excluding most other points. When applied to speech signals, vocal tract resonances (formants) or glottal pulsations can be effectively isolated in expanded views of the phonation process. PMID- 17407889 TI - A Bayesian method to estimate the depth and the range of phonating sperm whales using a single hydrophone. AB - Some bioacousticians have used a single hydrophone to calculate the depth/range of phonating diving animals. The standard one-hydrophone localization method uses multipath transmissions (direct path, sea surface, and seafloor reflections) of the animal phonations as a substitute for a vertical hydrophone array. The standard method requires three multipath transmissions per phonation. Bioacousticians who study foraging sperm whales usually do not have the required amount of multipath transmissions. However, they usually detect accurately (using shallow hydrophones towed by research vessels) direct path transmissions and sea surface reflections of sperm whale phonations (clicks). Sperm whales emit a few thousand clicks per foraging dive, therefore researchers have this number of direct path transmissions and this number of sea surface reflections per dive. The author describes a Bayesian method to combine the information contained in those acoustic data plus visual observations. The author's tests using synthetic data show that the accurate estimation of the depth/range of sperm whales is possible using a single hydrophone and without using any seafloor reflections. This method could be used to study the behavior of sperm whales using a single hydrophone in any location no matter what the depth, the relief, or the constitution of the seafloor might be. PMID- 17407890 TI - Toward bias minimization in acoustic feedback cancellation systems. AB - A novel technique for bias suppression within acoustic feedback cancellation systems is proposed. This is achieved based on the use of all-pass filters in the forward part of the hearing aid. The poles of these filters are made time varying, which results in a frequency response with constant magnitude and varying phase. This is a desired feature of the proposed approach, since the results from human psychoacoustics show that the human ear is not sensitive to moderate phase perturbations. The derivation of the proposed algorithms for the time variation of the location of the poles of all pass filters is based on a rigorous analysis of the phenomenon of bias in acoustic systems. Practical issues, such as the dependence of the steady-state error on the order of the all pass filter, the number of varying poles, and their standard deviation are examined and strategies for the variation of the poles are introduced. Results obtained from a simulated hearing aid are provided to support the analysis. The quality of the processed audio signals is evaluated through subjective tests. PMID- 17407891 TI - A full vectorial contrast source inversion scheme for three-dimensional acoustic imaging of both compressibility and density profiles. AB - Imaging the two acoustic medium parameters density and compressibility requires the use of both the acoustic pressure and velocity wave fields, described via integral equations. Imaging is based on solving for the unknown medium parameters using known measured scattered wave fields, and it is difficult to solve this ill posed inverse problem directly using a conjugate gradient inversion scheme. Here, a contrast source inversion method is used in which the contrast sources, defined via the product of changes in compressibility and density with the pressure and velocity wave fields, respectively, are computed iteratively. After each update of the contrast sources, an update of the medium parameters is obtained. Total variation as multiplicative regularization is used to minimize blurring in the reconstructed contrasts. The method successfully reconstructed three-dimensional contrast profiles based on changes in both density and compressibility, using synthetic data both with and without 50% white noise. The results were compared with imaging based only on the pressure wave field, where speed of sound profiles were solely based on changes in compressibility. It was found that the results improved significantly by using the full vectorial method when changes in speed of sound depended on changes in both compressibility and density. PMID- 17407892 TI - Statistically optimized near field acoustic holography using an array of pressure velocity probes. AB - Statistically optimized near field acoustic holography (SONAH) differs from conventional near field acoustic holography (NAH) by avoiding spatial Fourier transforms; the processing is done directly in the spatial domain. The main advantage of SONAH compared with NAH is that the usual requirement of a measurement aperture that extends well beyond the source can be relaxed. Both NAH and SONAH are based on the assumption that all sources are on one side of the measurement plane whereas the other side is source free. An extension of the SONAH procedure based on measurement with a double layer array of pressure microphones has been suggested. The double layer technique makes it possible to distinguish between sources on the two sides of the array and thus suppress the influence of extraneous noise coming from the "wrong" side. It has also recently been demonstrated that there are significant advantages in NAH based on an array of acoustic particle velocity transducers (in a single layer) compared with NAH based on an array of pressure microphones. This investigation combines the two ideas and examines SONAH based on an array of pressure-velocity intensity probes through computer simulations as well as experimentally. PMID- 17407893 TI - Nearfield binaural synthesis and ambisonics. AB - Ambisonic encodings can be rendered binaurally, as well as for speaker arrays. This process is developed for general high-order Ambisonic encodings of soundfields containing near as well as far sources. For sufficently near sources an error is identified, resulting from the limited field of validity of the freefield harmonic expansion. A modified expansion is derived that can render such sources correctly. PMID- 17407894 TI - Allen-Fahey and related experiments support the predominance of cochlear slow wave otoacoustic emissions. AB - Originally proposed as a method for measuring the power gain of the cochlear amplifier, Allen-Fahey experiments compare intracochlear distortion products and ear-canal otoacoustic emissions (OAEs) under tightly controlled conditions. In this paper Allen-Fahey experiments are shown to place significant constraints on the dominant mode of reverse energy propagation within the cochlea. Existing Allen-Fahey experiments are reviewed and shown to contradict the predictions of compression-wave OAE models recently proposed in the literature. In compression wave models, distortion products propagate from their site of generation to the stapes via longitudinal compression waves in the cochlear fluids (fast waves); in transverse traveling-wave models, by contrast, distortion products propagate primarily via pressure-difference waves whose velocity and other characteristics depend on the mechanical properties of the cochlear partition (slow waves). Compression-wave models predict that the distortion-product OAEs (DPOAEs) measured in the Allen-Fahey paradigm increase at close primary-frequency ratios (or remain constant in the hypothetical absence of tuned suppression). The behavior observed experimentally is just the opposite-a pronounced decrease in DPOAE amplitude at close ratios. Since neither compression-wave nor simple conceptual "hybrid-wave" models can account for the experimental results--whereas slow-wave models can, via systematic changes in distortion-source directionality arising from wave-interference effects--Allen-Fahey and related experiments provide compelling evidence against the predominance of compression-wave OAEs in mammalian cochlear mechanics. PMID- 17407895 TI - Examination of bone-conducted transmission from sound field excitation measured by thresholds, ear-canal sound pressure, and skull vibrations. AB - Bone conduction (BC) relative to air conduction (AC) sound field sensitivity is here defined as the perceived difference between a sound field transmitted to the ear by BC and by AC. Previous investigations of BC-AC sound field sensitivity have used different estimation methods and report estimates that vary by up to 20 dB at some frequencies. In this study, the BC-AC sound field sensitivity was investigated by hearing threshold shifts, ear canal sound pressure measurements, and skull bone vibrations measured with an accelerometer. The vibration measurement produced valid estimates at 400 Hz and below, the threshold shifts produced valid estimates at 500 Hz and above, while the ear canal sound pressure measurements were found erroneous for estimating the BC-AC sound field sensitivity. The BC-AC sound field sensitivity is proposed, by combining the present result with others, as frequency independent at 50 to 60 dB at frequencies up to 900 Hz. At higher frequencies, it is frequency dependent with minima of 40 to 50 dB at 2 and 8 kHz, and a maximum of 50 to 60 dB at 4 kHz. The BC-AC sound field sensitivity is the theoretical limit of maximum attenuation achievable with ordinary hearing protection devices. PMID- 17407896 TI - Bias due to noise in otoacoustic emission measurements. AB - Measurements of otoacoustic emission (OAE) magnitude are often made at low signal/noise ratios (SNRs) where measurement noise generates bias and variability errors that have led to the misinterpretation of OAE data. To gain an understanding for these errors and their effects, a two part investigation was carried out. First, the nature of OAE measurement noise was investigated using human data from 50 stimulus-frequency OAE experiments involving medial olivocochlear reflex (MOCR) activation. The noise was found to be reasonably approximated by circular Gaussian noise. Furthermore, when bias errors were taken into account, measurement variability was not found to be affected by MOCR activation as had been previously reported. Second, to quantify the errors circular Gaussian noise produces for different methods of OAE magnitude estimation for distortion-product, stimulus-frequency, and spontaneous OAEs, simulated OAE measurements were analyzed via four different magnitude estimation methods and compared. At low SNRs (below -6 dB), estimators involving Rice probability density functions produced less biased estimates of OAE magnitudes than conventional estimation methods, and less total rms error-particularly for spontaneous OAEs. They also enabled the calculation of probability density functions for OAE magnitudes from experimental data. PMID- 17407897 TI - Modeling binaural loudness. AB - A survey of data on the perception of binaurally presented sounds indicates that loudness summation across ears is less than perfect; a diotic sound is less than twice as loud as the same sound presented monaurally. The loudness model proposed by Moore et al. [J. Audio Eng. Soc. 45, 224-240 (1997)] determines the loudness of binaural stimuli by a simple summation of loudness across ears. It is described here how the model can be modified so as to give more accurate predictions of the loudness of binaurally presented sounds, including cases where the sounds at the two ears differ in level, frequency or both. The modification is based on the idea that there are inhibitory interactions between the internal representations of the signals at the two ears, such that a signal at the left ear inhibits (reduces) the loudness evoked by a signal at the right ear, and vice versa. The inhibition is assumed to spread across frequency channels. The modified model gives reasonably accurate predictions of a variety of data on the loudness of binaural stimuli, including data obtained using loudness scaling and loudness matching procedures. PMID- 17407898 TI - Hearing loss from interrupted, intermittent, and time varying Gaussian noise exposures: the applicability of the equal energy hypothesis. AB - Eight groups of chinchillas (N=74) were exposed to various equivalent energy [100 or 106 dB(A) sound pressure level (SPL)] noise exposure paradigms. Six groups received an interrupted, intermittent, time varying (IITV) Gaussian noise exposure that lasted 8 h/d, 5 d/week for 3 weeks. The exposures modeled an idealized workweek. At each level, three different temporal patterns of Gaussian IITV noise were used. The 100 dB(A) IITV exposure had a dB range of 90-108 dB SPL while the range of the 106 dB(A) IITV exposure was 80-115 dB SPL. Two reference groups were exposed to a uniform 100 or 106 dB(A) SPL noise, 24 h/d for 5 days. Each reference group and the three corresponding IITV groups comprised a set of equivalent energy exposures. Evoked potentials were used to estimate hearing thresholds and surface preparation histology quantified sensory cell populations. All six groups exposed to the IITV noise showed threshold toughening effects of up to 40 dB. All IITV exposures produced hearing and sensory cell loss that was similar to their respective equivalent energy reference group. These results indicate that for Gaussian noise the equal energy hypothesis for noise-induced hearing loss is an acceptable unifying principle. PMID- 17407899 TI - Across-frequency pitch discrimination interference between complex tones containing resolved harmonics. AB - Pitch discrimination interference (PDI) refers to an impairment in the ability to discriminate changes in the fundamental frequency (F0) of a target harmonic complex, caused by another harmonic complex (the interferer) presented simultaneously in a remote spectral region. So far, PDI has been demonstrated for target complexes filtered into a higher spectral region than the interferer and containing no peripherally resolved harmonics in their passband. Here, it is shown that PDI also occurs when the target harmonic complex contains resolved harmonics in its passband (experiment 1). PDI was also observed when the target was filtered into a lower spectral region than that of the interferer (experiment 2), revealing that differences in relative harmonic dominance and pitch salience between the simultaneous target and the interferer, as confirmed using pitch matches (experiment 3), do not entirely explain PDI. When the target was in the higher spectral region, and the F0 separation between the target and the interferer was around 7% or 10%, dramatic PDI effects were observed despite the relatively large FO separation between the two sequential targets (14%-20%). Overall, the results suggest that PDI is more general than previously thought, and is not limited to targets consisting only of unresolved harmonics. PMID- 17407900 TI - Illusory percepts from auditory adaptation. AB - Phenomena resembling tinnitus and Zwicker phantom tone are seen to result from an auditory gain adaptation mechanism that attempts to make full use of a fixed capacity channel. In the case of tinnitus, the gain adaptation enhances internal noise of a frequency band otherwise silent due to damage. This generates a percept of a phantom sound as a consequence of hearing loss. In the case of Zwicker tone, a frequency band is temporarily silent during the presentation of a notched broadband sound, resulting in a percept of a tone at the notched frequency. The model suggests a link between tinnitus and the Zwicker tone percept, in that it predicts different results for normal and tinnitus subjects due to a loss of instantaneous nonlinear compression. Listening experiments on 44 subjects show that tinnitus subjects (11 of 44) are significantly more likely to hear the Zwicker tone. This psychoacoustic experiment establishes the first empirical link between the Zwicker tone percept and tinnitus. Together with the modeling results, this supports the hypothesis that the phantom percept is a consequence of a central adaptation mechanism confronted with a degraded sensory apparatus. PMID- 17407901 TI - Threshold and channel interaction in cochlear implant users: evaluation of the tripolar electrode configuration. AB - The efficacy of cochlear implants is limited by spatial and temporal interactions among channels. This study explores the spatially restricted tripolar electrode configuration and compares it to bipolar and monopolar stimulation. Measures of threshold and channel interaction were obtained from nine subjects implanted with the Clarion HiFocus-I electrode array. Stimuli were biphasic pulses delivered at 1020 pulses/s. Threshold increased from monopolar to bipolar to tripolar stimulation and was most variable across channels with the tripolar configuration. Channel interaction, quantified by the shift in threshold between single- and two-channel stimulation, occurred for all three configurations but was largest for the monopolar and simultaneous conditions. The threshold shifts with simultaneous tripolar stimulation were slightly smaller than with bipolar and were not as strongly affected by the timing of the two channel stimulation as was monopolar. The subjects' performances on clinical speech tests were correlated with channel-to-channel variability in tripolar threshold, such that greater variability was related to poorer performance. The data suggest that tripolar channels with high thresholds may reveal cochlear regions of low neuron survival or poor electrode placement. PMID- 17407902 TI - Quantifying the effects of fast-acting compression on the envelope of speech. AB - Fast-acting compression has several effects on the envelope of speech signals, especially when a target and a background are mixed together. Three measures of the envelope are described, which can be used to quantify these changes: (1) Within-signal modulation correlation or coherence, the degree of correlation (or coherence) of the envelope (on a dB scale) of a single source across different frequency regions, which is reduced by fast-acting compression; (2) fidelity of envelope shape, the degree to which the envelope shape of the target speech in different frequency channels is preserved following compression; and (3) across signal modulation correlation or coherence (ASMC), the extent to which the target and background acquire a common component of modulation when they are compressed together, which becomes greater in absolute value (more negative) when the target and background are compressed together. Values of these measures are presented and compared with intelligibility scores obtained using stimuli processed (with a noise-vocoder) so as to preserve mainly envelope cues in a limited number of frequency channels. The results suggest that the dominant factor affecting intelligibility is ASMC. PMID- 17407903 TI - Word-internal versus word-peripheral consonantal duration patterns in three languages. AB - Segmental duration patterns have long been used to support the proposal that syllables are basic speech planning units, but production experiments almost always confound syllable and word boundaries. The current study tried to remedy this problem by comparing word-internal and word-peripheral consonantal duration patterns. Stress and sequencing were used to vary the nominal location of word internal boundaries in American English productions of disyllabic nonsense words with medial consonant sequences. The word-internal patterns were compared to those that occurred at the edges of words, where boundary location was held constant and only stress and sequence order were varied. The English patterns were then compared to patterns from Russian and Finnish. All three languages showed similar effects of stress and sequencing on consonantal duration, but an independent effect of syllable position was observed only in English and only at a word boundary. English also showed stronger effects of stress and sequencing across a word boundary than within a word. Finnish showed the opposite pattern, whereas Russian showed little difference between word-internal and word peripheral patterns. Overall, the results suggest that the suprasegmental units of motor planning are language-specific and that the word may be more a relevant planning unit in English. PMID- 17407904 TI - Noise estimation in voice signals using short-term cepstral analysis. AB - Cepstral-based estimation is used to provide a baseline estimate of the noise level in the logarithmic spectrum for voiced speech. A theoretical description of cepstral processing of voiced speech containing aspiration noise, together with supporting empirical data, is provided in order to illustrate the nature of the noise baseline estimation process. Taking the Fourier transform of the liftered (filtered in the cepstral domain) cepstrum produces a noise baseline estimate. It is shown that Fourier transforming the low-pass liftered cepstrum is comparable to applying a moving average (MA) filter to the logarithmic spectrum and hence the baseline receives contributions from the glottal source excited vocal tract and the noise excited vocal tract. Because the estimation process resembles the action of a MA filter, the resulting noise baseline is determined by the harmonic resolution (as determined by the temporal analysis window length) and the glottal source spectral tilt. On selecting an appropriate temporal analysis window length the estimated baseline is shown to lie halfway between the glottal excited vocal tract and the noise excited vocal tract. This information is employed in a new harmonics-to-noise (HNR) estimation technique, which is shown to provide accurate HNR estimates when tested on synthetically generated voice signals. PMID- 17407905 TI - Effect of spectral frequency range and separation on the perception of asynchronous speech. AB - The use of across-frequency timing cues and the effect of disrupting these cues were examined across the frequency spectrum by introducing between-band asynchronies to pairs of narrowband temporal speech patterns. Sentence intelligibility by normal-hearing listeners fell when as little as 12.5 ms of asynchrony was introduced and was reduced to floor values by 100 ms. Disruptions to across-frequency timing had similar effects in the low-, mid-, and high frequency regions, but band pairs having wider frequency separation were less disrupted by small amounts of asynchrony. In experiment 2, it was found that the disruptive influence of asynchrony on adjacent band pairs did not result from disruptions to the complex patterns present in overlapping excitation. The results of experiment 3 suggest that the processing of speech patterns may take place using mechanisms having different sensitivities to exact timing, similar to the dual mechanisms proposed for within- and across-channel gap detection. Preservation of relative timing can be critical to intelligibility. While the use of across-frequency timing cues appears similar across the spectrum, it may differ based on frequency separation. This difference appears to involve a greater reliance on exact timing during the processing of speech energy at proximate frequencies. PMID- 17407906 TI - Effects of stimulus variability and adult aging on adaptation to time-compressed speech. AB - With as few as 10-20 sentences of exposure, listeners are able to adapt to speech that is highly distorted compared to that which is encountered in everyday conversation. The current study examines the extent to which adaptation to time compressed speech can be impeded by disrupting the continuity of the exposure sentences, and whether this differs between young and older adult listeners when they are equated for starting accuracy. In separate sessions conducted one week apart, the degree of adaptation was assessed in four exposure conditions, all of which involved exposure to the same number of time-compressed sentences. A continuous exposure condition involved presentation of the time-compressed sentences without interruption. Two alternation conditions alternated time compressed speech and uncompressed speech by single sentences or groups of four sentences. A fourth condition presented sentences that were separated by a period of silence but no uncompressed speech. For all conditions, neither young nor older adults' overall level of learning was influenced by disruptions to the exposure sentences. In addition, participants' performance showed reliable improvement across the first and subsequent sessions. These results support robust learning mechanisms in speech perception that remain functional throughout the lifespan. PMID- 17407907 TI - Evaluation of companding-based spectral enhancement using simulated cochlear implant processing. AB - This study tested a time-domain spectral enhancement algorithm that was recently proposed by Turicchia and Sarpeshkar [IEEE Trans. Speech Audio Proc. 13, 243-253 (2005)]. The algorithm uses a filter bank, with each filter channel comprising broadly tuned amplitude compression, followed by more narrowly tuned expansion (companding). Normal-hearing listeners were tested in their ability to recognize sentences processed through a noise-excited envelope vocoder that simulates aspects of cochlear-implant processing. The sentences were presented in a steady background noise at signal-to-noise ratios of 0, 3, and 6 dB and were either passed directly through an envelope vocoder, or were first processed by the companding algorithm. Using an eight-channel envelope vocoder, companding produced small but significant improvements in speech reception. Parametric variations of the companding algorithm showed that the improvement in intelligibility was robust to changes in filter tuning, whereas decreases in the time constants resulted in a decrease in intelligibility. Companding continued to provide a benefit when the number of vocoder frequency channels was increased to sixteen. When integrated within a sixteen-channel cochlear-implant simulator, companding also led to significant improvements in sentence recognition. Thus, companding may represent a readily implementable way to provide some speech recognition benefits to current cochlear-implant users. PMID- 17407908 TI - Sinusoidal modeling for nonstationary voiced speech based on a local vector transform. AB - A voiced speech signal can be expressed as a sum of sinusoidal components of which instantaneous frequency and amplitude continuously vary with time. Determining these parameters from the input, the time-varying characteristics are crucial error sources for the algorithms, which assume their stationarity within a local analysis segment. To overcome this problem, a new method is proposed, local vector transform (LVT), which can determine instantaneous frequency and amplitude for nonstationary sinusoids. The method does not assume the local stationarity. The effectiveness of LVT was examined in parameter determination for synthesized and naturally uttered speech signals. The instantaneous frequency for the first harmonic component was determined with an accuracy almost equal to that of the time-corrected instantaneous frequency method and higher accuracy than that of spectral peak-picking, autocorrelation, and cepstrum. The instantaneous amplitude was also determined accurately by LVT while considerable errors were left in the other algorithms. The signal reconstructed from the determined parameters by LVT agreed well with the corresponding component of voiced speech. These results suggest that the method is effective for analyzing time-varying voiced speech signals. PMID- 17407909 TI - Sound pressure level and spectral balance linearity and symmetry in the messa di voce of female classical singers. AB - The messa di voce, in its pure form a crescendo and decrescendo on one note, has been revered for centuries in classical singing, but the pedagogical assumptions of linearity and symmetry have received little critical assessment, especially across a wide fundamental frequency (F0) range. Five trained female classical singers performed a total of 318 messe di voce across their musical F0 range to identify its acoustical characteristics and the influence of F0. Sound pressure level (SPL) range was generally greater during crescendo at higher F0's and during decrescendo at lower FO's. Change in SPL during the messa di voce was predominantly nonlinear, and the shape of the SPL traces differed greatly between crescendo and decrescendo. Nonlinearity in SPL change was not related to SPL range but did show a F0 influence in decrescendo. Change in spectral balance (0-2 vs. 2-4 kHz) with respect to SPL change showed markedly more symmetry than linearity, so that changes in the mode of phonation during the messa di voce were dependent upon SPL regardless of whether the singer was in crescendo or decrescendo. Perceptual and physiological implications are discussed. PMID- 17407910 TI - Gaussian mixture model classification of odontocetes in the Southern California Bight and the Gulf of California. AB - A method for the automatic classification of free-ranging delphinid vocalizations is presented. The vocalizations of short-beaked and long-beaked common (Delphinus delphis and Delphinus capensis), Pacific white-sided (Lagenorhynchus obliquidens), and bottlenose (Tursiops truncatus) dolphins were recorded in a pelagic environment of the Southern California Bight and the Gulf of California over a period of 4 years. Cepstral feature vectors are extracted from call data which contain simultaneous overlapping whistles, burst-pulses, and clicks from a single species. These features are grouped into multisecond segments. A portion of the data is used to train Gaussian mixture models of varying orders for each species. The remaining call data are used to test the performance of the models. Species are predicted based upon probabilistic measures of model similarity with test segment groups having durations between 1 and 25 s. For this data set, 256 mixture Gaussian mixture models and segments of at least 10 s of call data resulted in the best classification results. The classifier predicts the species of groups with 67%-75% accuracy depending upon the partitioning of the training and test data. PMID- 17407911 TI - Echo-intensity compensation in echolocating bats (Pipistrellus abramus) during flight measured by a telemetry microphone. AB - An onboard microphone (Telemike) was developed to examine changes in the basic characteristics of echolocation sounds of small frequency-modulated echolocating bats, Pipistrellus abramus. Using a dual high-speed video camera system, spatiotemporal observations of echolocation characteristics were conducted on bats during a landing flight task in the laboratory. The Telemike allowed us to observe emitted pulses and returning echoes to which the flying bats listened during flight, and the acoustic parameters could be precisely measured without traditional problems such as the directional properties of the recording microphone and the emitted pulse, or traveling loss of the sound in the air. Pulse intensity in bats intending to land exhibited a marked decrease by 30 dB within 2 m of the target wall, and the reduction rate was approximately 6.5 dB per halving of distance. The intensity of echoes returning from the target wall indicated a nearly constant intensity (-42.6 +/- 5.5 dB weaker than the pulse emitted in search phase) within a target distance of 2 m. These findings provide direct evidence that bats adjust pulse intensity to compensate for changes in echo intensity to maintain a constant intensity of the echo returned from the approaching target at an optimal range. PMID- 17407912 TI - Nonlinear acoustics in the pant hoots of common chimpanzees (Pan troglodytes): vocalizing at the edge. AB - Common chimpanzee (Pan troglodytes) "pant hoots" are multi-call events that build from quiet, consistently harmonic introductory sounds to loud, screamlike "climax" calls with acoustic irregularities known as "nonlinear phenomena" (NLP). Two possible functions of NLP in climax calls are to increase direct auditory impact on listeners and to signal physical condition. These possibilities were addressed by comparing climax calls from 12 wild chimpanzee males with "screams" and pant hoot "introduction" calls from the same individuals. Climax calls that included NLP were found to have higher fundamental frequencies (F0s) than introduction or climax calls that were purely harmonic. NLP onsets within climax calls were also specifically associated with local F0 maxima, suggesting vocalizers are vibrating their vocal folds at the upper limits of stability. Furthermore, climax calls showed far fewer NLP than did screams recorded from the same individuals, while showing equivalent or higher F0 values. Overall, the results are consistent with the hypothesis that the relative prevalence of NLP is a signal of physical condition, with callers "vocalizing at the edge" of regular, stable production while producing few NLP. The results are discussed in light of the initial hypotheses. PMID- 17407913 TI - The perception of complex tones by a false killer whale (Pseudorca crassidens). AB - Complex tonal whistles are frequently produced by some odontocete species. However, no experimental evidence exists regarding the detection of complex tones or the discrimination of harmonic frequencies by a marine mammal. The objectives of this investigation were to examine the ability of a false killer whale to discriminate pure tones from complex tones and to determine the minimum intensity level of a harmonic tone required for the whale to make the discrimination. The study was conducted with a go/no-go modified staircase procedure. The different stimuli were complex tones with a fundamental frequency of 5 kHz with one to five harmonic frequencies. The results from this complex tone discrimination task demonstrated: (1) that the false killer whale was able to discriminate a 5 kHz pure tone from a complex tone with up to five harmonics, and (2) that discrimination thresholds or minimum intensity levels exist for each harmonic combination measured. These results indicate that both frequency level and harmonic content may have contributed to the false killer whale's discrimination of complex tones. PMID- 17407914 TI - Bottlenose dolphin (Tursiops truncatus) steady-state evoked responses to multiple simultaneous sinusoidal amplitude modulated tones. AB - Auditory steady-state evoked potentials were measured in a bottlenose dolphin (Tursiops truncatus) in response to single and multiple sinusoidal amplitude modulated (SAM) tones. Tests were conducted in air using a "jawphone" sound projector. Evoked potentials were recorded noninvasively using surface electrodes embedded in suction cups. Sound stimuli consisted of SAM tones with 1, 2, 3, or 4 carrier frequencies (10, 20, 30, 40 kHz), each with a unique modulation frequency. Stimulus sound pressure levels were varied in 5-dB steps from approximately 120 to 60-75 dB re 1 microPa, depending on frequency. Evoked potentials followed the temporal envelope of each stimulus, resulting in spectral components at each unique modulation frequency. Spectral analysis was used to evaluate the response amplitude for each carrier as a function of stimulus level. There were no significant differences between thresholds obtained with single and multiple stimuli at 10, 30, and 40 kHz. At 20 kHz, thresholds obtained with three components were higher than those obtained with four components, possibly revealing interactions between stimuli with less than one octave frequency separation. The use of multiple SAM stimuli may offer substantial advantages for studies of marine mammal hearing, where testing time and access to subjects are typically limited. PMID- 17407915 TI - Comodulation detection differences in the hooded crow (Corvus corone cornix), with direct comparison to human subjects. AB - Envelope modulations have been shown important in determining the effectiveness of masking noises. For example, the threshold for detecting a signal flanked by maskers is lower if the maskers and the signal are modulated with different envelopes, rather than the same envelope (comodulation). This threshold change is called the comodulation detection difference (CDD). CDDs were studied in two wild caught hooded crows, using a 1.5 kHz signal and two maskers at 0.9 and 2.1 kHz, presented at an overall level of 55 dB SPL (re 20 microPa). For direct comparison with human psychophysics, three human subjects were tested in the same setup. CDDs averaged 15 dB for the two crow subjects and 11 dB for the human subjects. The species difference between average CDDs was insignificant. The significance of the CDD effect in a natural setting is discussed. PMID- 17407916 TI - Singular spectrum analysis applied to ultrasonic detection and imaging of brachytherapy seeds. AB - Ultrasound-guided brachytherapy using titanium-shelled radioactive seeds is a popular, effective means of treating prostate cancer. Unfortunately, implantation using needles inserted transperitoneally causes gland movement and distortion, which often results in seed misplacement and dosimetry errors. If actual seed locations could be determined in the operating room, then corrections to dosimetry errors could be made immediately. However, seed specularity, shadowing, and tissue clutter make imaging seeds difficult using conventional ultrasound. Singular spectrum analysis (SSA) shows promise for reliably imaging radioactive seeds implanted in the prostate and enabling additional corrective implantations to be made in the operating room. SSA utilizes eigenvalues derived from the diagonalized correlation matrix of envelope-detected radio-frequency echo signals to yield a P value indicative of the likelihood of a seed-specific repetitive signal. We demonstrated the potential of SSA for seed detection and imaging and illustrated the trade-off considerations for optimization of SSA in clinical applications using simulations assessing performance as a function of different levels of noise and the presence of repetitive signals with various repetition periods; experiments in an ideal scattering environment; and experiments using seeds implanted in beef. PMID- 17407917 TI - Interaural time difference processing of broadband and narrow-band noise by inexperienced listeners. AB - Recent functional magnetic resonance imaging (fMRI) data might be interpreted as being in disagreement with existing psychophysical data regarding the laterality of broadband noise stimuli presented with large interaural time differences (ITDs). This study investigated the possibility that lateral judgments made by inexperienced listeners who did not receive feedback might be different than those reported for experienced listeners, especially when the ITD is longer than that occurring in nature, and therefore data from inexperienced listeners presented unnaturally long ITDs for the first time might be more consistent with the possible interpretation of the fMRI results. The results from this study using inexperienced listeners were not basically different from those reported in the literature based on experienced listeners, suggesting a possible difference does exist between inferences drawn from fTMRI data and human psychophysical results. PMID- 17407918 TI - Acoustic micro-Doppler radar for human gait imaging. AB - A portable acoustic micro-Doppler radar system for the acquisition of human gait signatures in indoor and outdoor environments is reported. Signals from an accelerometer attached to the leg support the identification of the components in the measured micro-Doppler signature. The acoustic micro-Doppler system described in this paper is simpler and offers advantages over the widely used electromagnetic wave micro-Doppler radars. PMID- 17407919 TI - Ultrasonic wave generation due to human footsteps on the ground. AB - Human footsteps generate broadband frequency vibrations in the ground/floor and sound in the air from a few Hertz up to ultrasonic frequencies due to striking and sliding contacts between a foot and the ground/floor. The high-frequency (above 1 kHz) vibrations from footsteps were detected on a building floor, but were not detected on the outdoor ground, even at 1 m from a walker. This paper presents results of ultrasound registration from footsteps on the ground at greater distances. Results are based on sound measurements in air, since the sound absorption in air is less than vibration absorption in the ground. PMID- 17407920 TI - Acoustic measures of low-frequency noise in extended high-frequency audiometry. AB - A very high signal-to-noise ratio is required for equipment designed for extended high-frequency audiometry because listeners with almost no hearing ability in the extended high-frequency range may have normal hearing sensitivity in the lower frequencies. Two commercially available systems designed for pure-tone audiometry were evaluated both in the conventional and extended high-frequency range. Unwanted lower frequency signals greater than the noise floor occurred predominantly at presentation levels of approximately 110 dB SPL or higher. Test tones in the extended high-frequency range should be restricted to levels that are not associated with lower frequency noise. PMID- 17407921 TI - What is the problem? The challenge of providing effective teachers for all children. AB - Richard Murnane and Jennifer Steele argue that if the United States is to equip its young people with the skills essential in the new economy, high-quality teachers are more important than ever. In recent years, the demand for effective teachers has increased as enrollments have risen, class sizes have fallen, and a large share of the teacher workforce has begun to retire. Women and minorities have more career options than ever before, making it increasingly difficult to attract and retain the many effective teachers who are needed. Moreover, schools are limited in their ability to identify and reward the most effective teachers. Perhaps the most urgent problem facing American education, say Murnane and Steele, is the unequal distribution of high-quality teachers. Poor children and children of color are disproportionately assigned to teachers with the least preparation and the weakest academic backgrounds. Teacher turnover is high in schools that serve large shares of poor or nonwhite students because the work is difficult, and the teachers who undertake it are often the least equipped to succeed. Murnane and Steele point out that in response to these challenges, policymakers have proposed a variety of policy instruments to increase the supply of effective teachers and distribute those teachers more equitably across schools. Such proposals include across-the-board pay increases, more flexible pay structures such as pay-for-performance, and reduced restrictions on who is allowed to teach. Several of these proposals are already being implemented, but their effectiveness remains largely unknown. To measure how well these policies attract effective teachers to the profession and to the schools that need them most, rigorous evaluations are essential. Murnane and Steele also note that policymakers may benefit from looking beyond U.S. borders to understand how teacher labor markets work in other countries. Although policies rooted in one nation's culture cannot be easily and quickly transplanted into another, it is important to understand what challenges other countries face, what policies they are using, and how well those policies are working to enhance teacher quality and improve student achievement. PMID- 17407922 TI - The effect of certification and preparation on teacher quality. AB - To improve the quality of the teacher workforce, some states have tightened teacher preparation and certification requirements while others have eased requirements and introduced "alternative" ways of being certified to attract more people to teaching. Donald Boyd, Daniel Goldhaber, Hamilton Lankford, and James Wyckoff evaluate these seemingly contradictory strategies by examining how preparation and certification requirements affect student achivement. If strong requirements improve student outcomes and deter relatively few potential teachers, the authors say, then they may well be good policy. But if they have little effect on student achievement, if they seriously deter potential teachers, or if schools are able to identify applicants who will produce good student outcomes, then easing requirements becomes a more attractive policy. In reviewing research on these issues, the authors find that evidence is often insufficient to draw conclusions. They do find that highly selective alternative route programs can produce effective teachers who perform about the same as teachers from traditional routes after two years on the job. And they find that teachers who score well on certification exams can improve student outcomes somewhat. Limited evidence suggests that certification requirements can diminish the pool of applicants, but there is no evidence on how they affect student outcomes. And the authors find that schools have a limited ability to identify attributes in prospective teachers that allow them to improve student achievement. The authors conclude that the research evidence is simply too thin to have serious implications for policy. Given the enormous investment in teacher preparation and certification and given the possibility that these requirements may worsen student outcomes, the lack of convincing evidence is disturbing. The authors urge researchers and policymakers to work together to move to a more informed position where good resource decisions can be made. PMID- 17407923 TI - Pay, working conditions, and teacher quality. AB - Eric Hanushek and Steven Rivkin examine how salary and working conditions affect the quality of instruction in the classroom. The wages of teachers relative to those of other college graduates have fallen steadily since 1940. Today, average wages differ little, however, between urban and suburban districts. In some metropolitan areas urban districts pay more, while in others, suburban districts pay more. But working conditions in urban and suburban districts differ substantially, with urban teachers reporting far less administrator and parental support, worse materials, and greater student problems. Difficult working conditions may drive much of the difference in turnover of teachers and the transfer of teachers across schools. Using rich data from Texas public schools, the authors describe in detail what happens when teachers move from school to school. They examine how salaries and student characteristics change when teachers move and also whether turnover affects teacher quality and student achievement. They note that both wages and student characteristics affect teachers' choices and result in a sorting of teachers across schools, but they find little evidence that teacher transitions are detrimental to student learning. The extent to which variations in salaries and working conditions translate into differences in the quality of instruction depends importantly on the effectiveness of school personnel policies in hiring and retaining the most effective teachers and on constraints on both entry into the profession and the firing of low performers. The authors conclude that overall salary increases for teachers would be both expensive and ineffective. The best way to improve the quality of instruction would be to lower barriers to becoming a teacher, such as certification, and to link compensation and career advancement more closely with teachers' ability to raise student performance. PMID- 17407924 TI - Using performance-based pay to improve the quality of teachers. AB - Tying teachers' pay to their classroom performance should, says Victor Lavy, improve the current educational system both by clarifying teaching goals and by attracting and retaining the most productive teachers. But implementing pay for performance poses many practical challenges, because measuring individual teachers' performance is difficult. Lavy reviews evidence on individual and school-based incentive programs implemented in recent years both in the United States and abroad. Lavy himself evaluated two carefully designed programs in Israel and found significant gains in student and teacher performance. He observes that research evidence suggests, although not conclusively, that pay-for performance incentives can improve teachers' performance, although they can also lead to unintended and undesired consequences, such as teachers' directing their efforts exclusively to rewarded activities. Lavy also offers general guidelines for designing effective programs. He emphasizes that the system must measure true performance in a way that minimizes random variation as well as undesired and unintended consequences. It must align performance with ultimate outcomes and must be monitored closely to discourage gaming if not outright fraud in measured output. Goals should be attainable. Incentives should balance individual rewards with school incentives, fostering a cooperative culture but not at the expense of free riding. All teachers should be eligible for the incentive offered, but only a subset of teachers should be rewarded in practice. If too many teachers are rewarded, teachers may not need to exert much extra effort to benefit. Many of the practical challenges faced by performance-related pay, Lavy says, can be addressed through careful design of the system. He emphasizes that setting up a performance-related pay system that works is not a one-time task. Even with the best preparation, initial implementation is likely to be problematic. But if the effort is seen as ongoing, it should be possible to make progress gradually in developing incentives that motivate the desired teaching behaviors and that will be perceived by teachers as fair and accurate. PMID- 17407925 TI - Learning in the teaching workforce. AB - The U.S. educational system invests heavily, in both time and money, in continuing education for teachers. In this article Heather Hill examines the effectiveness of two forms of teacher learning-graduate coursework and professional development. She focuses first on graduate education. Almost half of all teachers have a master's degree. Many states allow graduate coursework to count toward recertification requirements. Some districts require teachers to complete a master's degree within several years of hiring, and many others reward it with salary increases. Education reformers often recommend requiring master's degrees. But much graduate coursework appears to be of low intellectual quality and disconnected from classroom practice. Most research finds no link between teachers' graduate degrees and student learning unless the degree is in the teacher's primary teaching field. Hill then examines professional development. Most workshops, institutes, and study groups appear to be brief, superficial, and of marginal use in improving teaching. But it does not have to be this way, says Hill. Professional development can enhance teaching and learning if it has three characteristics. It must last several days or longer; it must focus on subject matter-specific instruction; and it must be aligned with the instructional goals and curriculum materials in teachers' schools. Such high-quality programs do exist. But they are a tiny fraction of the nation's offerings. One problem, says Hill, is that researchers rarely evaluate carefully either local professional development or its effect on student learning. Most evaluations simply ask participants to self-report. Lacking reliable evaluations, how are teachers and district officials to choose effective programs? Clearly, much more rigorous studies are needed. To make continuing education effective, school districts should encourage teachers to take graduate coursework that is more tightly aligned with their primary teaching assignment. And districts should select professional development programs based on evidence of their effectiveness. Finally, central planners must ensure that items on the menu of offerings closely align with district standards, curriculum materials, and assessments. PMID- 17407926 TI - The challenges of staffing urban schools with effective teachers. AB - Brian Jacob examines challenges faced by urban districts in staffing their schools with effective teachers. He emphasizes that the problem is far from uniform. Teacher shortages are more severe in certain subjects and grades than others, and differ dramatically from one school to another. The Chicago public schools, for example, regularly receive roughly ten applicants for each teaching position. But many applicants are interested in specific schools, and district officials struggle to find candidates for highly impoverished schools. Urban districts' difficulty in attracting and hiring teachers, says Jacob, means that urban teachers are less highly qualified than their suburban counterparts with respect to characteristics such as experience, educational background, and teaching certification. But they may not thus be less effective teachers. Jacob cites recent studies that have found that many teacher characteristics bear surprisingly little relationship to student outcomes. Policies to enhance teacher quality must thus be evaluated in terms of their effect on student achievement, not in terms of conventional teacher characteristics. Jacob then discusses how supply and demand contribute to urban teacher shortages. Supply factors involve wages, working conditions, and geographic proximity between teacher candidates and schools. Urban districts have tried various strategies to increase the supply of teacher candidates (including salary increases and targeted bonuses) and to improve retention rates (including mentoring programs). But there is little rigorous research evidence on the effectiveness of these strategies. Demand also has a role in urban teacher shortages. Administrators in urban schools may not recognize or value high-quality teachers. Human resource departments restrict district officials from making job offers until late in the hiring season, after many candidates have accepted positions elsewhere. Jacob argues that urban districts must improve hiring practices and also reevaluate policies for teacher tenure so that ineffective teachers can be dismissed. PMID- 17407927 TI - Recruiting and retaining high-quality teachers in rural areas. AB - In examining recruitment and retention of teachers in rural areas, David Monk begins by noting the numerous possible characteristics of rural communities- small size, sparse settlement, distance from population concentrations, and an economic reliance on agricultural industries that are increasingly using seasonal and immigrant workers to minimize labor costs. Many, though not all, rural areas, he says, are seriously impoverished. Classes in rural schools are relatively small, and teachers tend to report satisfaction with their work environments and relatively few problems with discipline. But teacher turnover is often high, and hiring can be difficult. Monk observes that rural schools have a below-average share of highly trained teachers. Compensation in rural schools tends to be low, perhaps because of a lower fiscal capacity in rural areas, thus complicating efforts to attract and retain teachers. Several student characteristics, including relatively large shares of students with special needs and with limited English skills and lower shares of students attending college, can also make it difficult to recruit and retain high-quality teachers. Other challenges include meeting the needs of highly mobile children of low-income migrant farm workers. With respect to public policy, Monk asserts a need to focus on a subcategory of what might be called hard-to-staff rural schools rather than to develop a blanket set of policies for all rural schools. In particular, he recommends a focus on such indicators as low teacher qualifications, teaching in fields far removed from the area of training, difficulty in hiring, high turnover, a lack of diversity among teachers in the school, and the presence of migrant farm workers' children. Successful efforts to stimulate economic growth in these areas would be highly beneficial. He also calls attention to the potential for modern telecommunication and computing technologies to offset some of the drawbacks associated with teaching in rural areas. PMID- 17407928 TI - Teachers unions and student performance: help or hindrance? AB - Randall Eberts explores the role of teachers unions in public education. He focuses particularly on how collective bargaining agreements shape the delivery of educational services, how unions affect both student achievement and the cost of providing quality education, and how they support educational reform efforts. Eberts's synthesis of the empirical research concludes that union bargaining raises teachers' compensation, improves their working conditions, and enhances their employment security-while also raising the cost of providing public education by upwards of 15 percent. The effect of unions on student performance is mixed. Students of average ability who attend school in union districts perform better on standardized tests, whereas low-achieving and high-achieving students perform worse. However, the overall gain in achievement does not make up for the higher cost. Of late, unions have begun to be more supportive of school reform, moving from an adversarial bargaining model to a more collaborative one in which teachers and administrators share common goals and hold joint responsibility. Yet unions' desire to participate in reform does not match their fervor to organize in the 1960s and 1970s. While national union leadership has talked about reform, local affiliates have initiated most of the reform efforts, pioneering reforms such as accountability and incentive pay. In Eberts's view, one reason that unions have been slow to embrace reform efforts is the lack of consensus on their effectiveness. He argues that many reforms have been too narrowly focused; rather, effective schools result from well-designed systems and processes. In principle, adopting standards that help teachers focus on lessons they want students to learn, aligning their teaching to the lessons, and devising measurements that demonstrate that students are responding to these lessons can improve teaching as long as the public, policymakers, and school administrators acknowledge the complexity of the learning process and the broad outcomes that society desires. PMID- 17407929 TI - Teacher labor markets in developed countries. AB - Helen Ladd takes a comparative look at policies that the world's industrialized countries are using to assure a supply of high-quality teachers. Her survey puts U.S. educational policies and practices into international perspective. Ladd begins by examining teacher salaries-an obvious, but costly, policy tool. She finds, perhaps surprisingly, that students in countries with high teacher salaries do not in general perform better on international tests than those in countries with lower salaries. Ladd does find, however, that the share of underqualified teachers in a country is closely related to salary. In high-salary countries like Germany, Japan, and Korea, for example, only 4 percent of teachers are underqualified, as against more than 10 percent in the United States, where teacher salaries, Ladd notes, are low relative to those in other industrialized countries. Teacher shortages also appear to stem from policies that make salaries uniform across academic subject areas and across geographic regions. Shortages are especially common in math and science, in large cities, and in rural areas. Among the policy strategies proposed to deal with such shortages is to pay teachers different salaries according to their subject area. Many countries are also experimenting with financial incentive packages, including bonuses and loans, for teachers in specific subjects or geographic areas. Ladd notes that many developed countries are trying to attract teachers by providing alternative routes into teaching, often through special programs in traditional teacher training institutions and through adult education or distance learning programs. To reduce attrition among new teachers, many developed countries have also been using formal induction or mentoring programs as a way to improve new teachers' chances of success. Ladd highlights the need to look beyond a single policy, such as higher salaries, in favor of broad packages that address teacher preparation and certification, working conditions, the challenges facing new teachers, and the distribution of teachers across geographic areas. PMID- 17407930 TI - Teacher labor markets in developing countries. AB - Emiliana Vegas surveys strategies used by the world's developing countries to fill their classrooms with qualified teachers. With their low quality of education and wide gaps in student outcomes, schools in developing countries strongly resemble hard-to-staff urban U.S. schools. Their experience with reform may thus provide insights for U.S. policymakers. Severe budget constraints and a lack of teacher training capacity have pushed developing nations to try a wide variety of reforms, including using part-time or assistant teachers, experimenting with pay incentives, and using school-based management. The strategy of hiring teachers with less than full credentials has had mixed results. One successful program in India hired young women who lacked teaching certificates to teach basic literacy and numeracy skills to children whose skills were seriously lagging. After two years, student learning increased, with the highest gains among the least able students. As in the United States, says Vegas, teaching quality and student achievement in the developing world are sensitive to teacher compensation. As average teacher salaries in Chile more than doubled over the past decade, higher-quality students entered teacher education programs. And when Brazil increased educational funding and distributed resources more equitably, school enrollment increased and the gap in student test scores narrowed. Experiments with performance-based pay have had mixed results. In Bolivia a bonus for teaching in rural areas failed to produce higher-quality teachers. And in Mexico a system to reward teachers for improved student outcomes failed to change teacher performance. But Vegas explains that the design of teacher incentives is critical. Effective incentive schemes must be tightly coupled with desired behaviors and generous enough to give teachers a reason to make the extra effort. School-based management reforms give decisionmaking authority to the schools. Such reforms in Central America have reduced teacher absenteeism, increased teacher work hours, increased homework assignments, and improved parent-teacher relationships. These changes, says Vegas, are especially promising in schools where educational quality is low. PMID- 17407931 TI - Navigation-guided transfer of cartilage defect geometry for arthroscopic autologous chondrocyte transplantation. AB - Cartilage repair with autologous chondrocyte transplantation shows intriguing results. Chondrogenic transplants generally must be inserted into cartilage defects via arthrotomy. The following study showed that arthroscopically-guided navigation could detect and precisely measure the cartilage defect sizes of different geometries. The new cartilage defect-managing module allowed for the precise transfer of navigated cartilage defect geometries for exact size preparation of the tissue engineering scaffolds. Therefore, navigation can help accomplish chondrocyte transplantation arthroscopically. PMID- 17407932 TI - Computer-assisted dynamic total knee arthroplasty using Whiteside's line for alignment. AB - Computer-assisted navigation enables surgeons to measure and assess knee behavior during surgery, allowing real-time monitoring of the knee ranging from extension to flexion. Documenting passive motion before and after total knee arthroplasty (TKA) enabled the authors to analyze knee kinematics. Passive knee motion cannot predict full weight bearing knee behavior, the "crucial moment of truth" for TKA. This study supports individually adapting the femoral component rotation using the Whiteside line. Clinical follow-up alone will assess the possible benefits. In a study of 71 patients who underwent computer-assisted TKA, the authors assessed the actual rotational femoral alignment and dynamic long leg track hip knee-ankle angle from 0 degrees, 30 degrees, 60 degrees, and 90 degrees of knee flexion before and after TKA. PMID- 17407933 TI - Future of double-bundle anterior cruciate ligament (ACL) reconstruction: incorporation of ACL anatomic data into the navigation system. AB - This study compares anatomy and tunnel placement during double-bundle procedures. Double-bundle anterior cruciate ligament (ACL) reconstruction reproduces both the anteromedial and posterolateral bundles. Thirty-six knees from cadavers were used for anatomic study, and 38 navigated double-bundle ACL reconstructions were evaluated for tunnel placements. With the exception of femoral tunnels of anteromedial bundles, all tunnel placements were in accordance with the anatomic measurements of the native ACL. The investigators concluded that incorporating ACL anatomic information into navigation systems would enhance double-bundle procedures. PMID- 17407934 TI - Current status of navigated cup replacement in dysplastic cases. AB - From the authors' experiences, several challenges exist in creating accurate cup positioning in patients with dysplastic hip due to anatomic abnormalities. B. Braun Aesculap (Tuttlingen, Germany) developed an advanced OrthoPilot software module for dysplastic cup positioning that has several advantages over the normal cup software as a solution to cup inaccuracies. The following study is a preliminary report on the new software version, which provides more specific functionality than the previous software system, a precondition or successful dysplastic cup navigation. This new functionality increase usefulness of navigation and the safety of the procedure. PMID- 17407935 TI - Navigated minimally invasive unicompartmental knee arthroplasty. AB - Unicompartmental knee arthroplasty (UKA) is an alternative procedure to high tibial osteotomy. This study assessed the procedure using computer navigation to improve implantation accuracy and presents early radiological results of a group of patients implanted with the univation UKA (B. Braun Aesculap, Tuttlingen, Germany) with navigation instrumentation and a minimally invasive approach. The authors concluded that navigated implantation of a UKA using a nonimage-based system improved radiologic accuracy implantation without significant inconvenience and minimal change in the conventional operating technique. PMID- 17407936 TI - Anterior cruciate ligament (ACL) tunnel placement: a radiographic comparison between navigated versus manual ACL reconstruction. AB - As participation in sports has increased, so have anterior cruciate ligament (ACL) reconstructions. The procedure is commonly associated with inaccurate tunnel placements, even when performed by experienced surgeons. Computer-assisted surgery may assist in eliminating inconsistent graft tunnels by increasing precision and providing navigation feedback of the surgical field. This study aimed to compare the accuracy of tunnel placements between computer-navigated and manual ACL reconstructions using radiography. Results suggest that the two methods have comparable accuracy but that the use of additional imaging techniques is recommended during tunnel placement evaluation. PMID- 17407937 TI - Navigation of short-stem implants. AB - Short-stem prostheses with modular necks were implanted using a modified Watson Jones approach in 55 cases from November 2004 to May 2006. Twenty-eight cases were navigated using the OrthoPilot navigation system (B. Braun Aesculap, Tuttlingen, Germany). Primary stability, ease of minimal invasive implantation, and restoration of the biomechanics were evaluated. Short-term results showed a good functional outcome and a low complication rate without any dislocations. Apart from cup navigation, the navigation system helps to restore biomechanics in terms of center of rotation, leg length, and offset by advising surgeons on the modular neck offering best fit predicting the safe range of motion (ROM) reliably. PMID- 17407938 TI - High tibial osteotomy: does navigation improve results? AB - Between January 2003 and March 2006, the authors performed 67 open-wedge high tibial osteotomies (HTO) for treatment of genu varum and osteoarthritis. Through comparison of two groups, the study sought to answer whether computed tomography (CT)-free navigation is feasible for HTO and will provide a more accurate correction angle without giving rise to additional complications. The results showed a significantly higher accuracy in achieving the proper leg axis correction (P< .016) when HTO was performed with navigation. Navigation in HTO, which allows intraoperative calculation of the leg axis, seems to be a reliable and safe procedure. Knowing that clinical results of HTO in varus deformity of the leg axis are closely connected to correct postoperative valgus alignment and avoiding overcorrection and undercorrection, the study lead to the conclusion that the use of navigation will contribute to better clinical outcomes. PMID- 17407939 TI - Navigated anterior cruciate ligament reconstruction: correlation between computer data and radiographic measurements. AB - The goal of this study was to prove the reliability of computer-aided navigation for the surgical reconstruction of the anterior cruciate ligament (ACL) using an arthroscopic procedure. The study involved 50 patients preceded by a learning curve period. Computer-recorded data were compared with specific radiography measurements of the frontal and anteroposterior views. The radiographs were used to measure the position of the tibial tunnel in the frontal and sagittal plane, the angulation of the tibial tunnel in the frontal and sagittal plane, and the position of the femoral tunnel in the lateral condyle. PMID- 17407940 TI - Navigation as a predictor of soft tissue release during 90 cases of computer assisted total knee arthroplasty. AB - Soft tissue balance during total knee arthroplasty (TKA) procedures may be challenging to achieve in knees with severe valgus or varus deformities. The purpose of this study was to evaluate the role of navigation in predicting soft tissue release during 90 computer-assisted TKA procedures performed within a 2 year period. Fourteen soft tissue releases were performed, 13 of which were for genu varum, including 9 deep medial collateral ligaments and 4 deep and superficial medial collateral ligaments, and 1 for genu valgum deformity. Forty one cases were intraoperatively overcorrectable (45.5%), and 20 cases were reducible to a neutral axis. The results proved that navigation can predict the need to perform a soft tissue release during computer-assisted TKA. PMID- 17407941 TI - Minimally invasive orthopedic surgery: first results in navigated total hip arthroplasty. AB - Minimally invasive hip approaches decrease soft tissue damage but reduce the view of the operating field. The combination of minimally invasive approaches with navigation techniques can resolve this conflict. A modified anterolateral approach was used for minimally invasive implantation of hip endoprostheses, in combination with a navigation technique. A newly developed femoral clamp was used in the study, which allowed a secure fixation of the navigation reference frame without compromising the visibility of the small incision. The results included short skin incisions and hip joint muscle reservation, with only a moderate increase in operating times. PMID- 17407942 TI - Minimally invasive computer-assisted total knee arthroplasty through a subvastus approach. AB - Computer-assisted total knee arthroplasty (TKA) has traditionally been performed through an anterior midline incision approximately 16 cm long, using a capsular incision that separates the interval between the rectus femoris and vastus medialis muscles. The incision disrupts the suprapatellar pouch and frequently leads to adhesions and difficulty in fast and complete flexion recovery despite the wide exposure. The minimally invasive surgery subvastus approach with computer-assisted soft tissue balancing using OrthoPilot TKA (B. Braun Aesculap, Tuttlingen, Germany) not only provides optimal alignment and balancing of the components but also results in maximal and faster flexion recovery and reduces the need for postoperative narcotic medication. PMID- 17407943 TI - Functional results of navigated minimally invasive and conventional total knee arthroplasty: a comparison in bilateral cases. AB - This prospective study aimed to compare the clinical and radiologic results achieved using navigation-assisted minimally invasive surgery (NA-MIS) and conventional techniques in 42 patients who underwent bilateral total knee arthroplasty (TKA), with a minimum 1-year follow-up. Patients who underwent NA MIS TKA showed better Hospital for Special Surgery (HSS) Scores and Western Ontario MacMaster (WOMAC) total scores for up to 6 months postoperatively than patients who underwent conventional TKA, and lower WOMAC pain score for up to 9 months postoperatively. Range of motion (ROM) was comparable in both groups at all times. More patients preferred the NA-MIS procedure to the conventional procedure. Radiologic results for the NA-MIS group showed fewer outliers than did results for the conventional group, but not in mean values. PMID- 17407944 TI - Rotational alignment of femoral components in total knee arthroplasty: nonimage based navigation system versus conventional technique. AB - A randomized prospective study was undertaken to compare the accuracy of rotational alignment of the femoral component in total knee arthroplasty using a nonimage-based navigation system and a conventional method. The authors randomly used two methods and implants in 55 patients and measured the angles between posterior condylar lines and transepicondylar axes on computed tomography images before and after surgery. Analysis showed no significant differences between the two techniques in mean errors (P>.05), but the navigation system showed higher accuracy than the 3 degrees external rotation method by reducing outliers. PMID- 17407945 TI - Blood loss after navigation-assisted minimally invasive total knee arthroplasty. AB - The following study evaluated blood loss in patients undergoing navigation assisted minimally invasive total knee arthroplasty. The study included 100 patients divided into two groups. Group A consisted of patients with an immediate drain release, and group B consisted of patients with a 1-hour delayed drain release after surgery. The surgeon used a mini-midvastus approach with the OrthoPilot navigation system. The mean blood drainage was 465 mL in group A and 409 mL in group B. The mean hemoglobin loss was 2.59 g/dL in group A and 2.43 g/dL in group B. PMID- 17407946 TI - [Anterior skull base reconstruction]. AB - The reasons for transcranial operative revision of anterior cranial fossa are cerebrospinal fluid (CSF) leakage, pneumocephalus, recurrent meningitis and eventually other causes. The trauma is the most common etiology of previously mentioned pathologic statuses, rarely it can also be long-lasting intracranial hypertension. We retrospectively analyzed our 61 patients, operatively treated at Department of Neurosurgery in Brno-Bohunice between 1997 and 2005, at whom the cranionasal communication must have been treated by means of transcranial neurosurgical repair of anterior fossa skull base. Each year approximately 10-15 operations of this kind were performed. The main goal of operative solution was the water-tight closure of dural defect and cranial base by means of direct dural suture duroplasty with periostal flap, fascial flap or other graft. Each case solution was dependent on actual operative appearance of the operative field. The majority of transcranial repairs of frontal cerebral fossas were performed acutely as a part of frontobasal injury operation with haematoma, calvar impressive fractures and open cerebral injury. The minority of repairs is formed by elective operations for posttraumatic rhinnorhea and meningitis. In our announcement we also mention possible conservative treatment. PMID- 17407947 TI - [Surgeries for blunt and penetrating thoracic injuries]. AB - INTRODUCTION: Up to 50% deaths due to fatal injuries result from thoracic injuries. Approximately 10-15 % of the injuries require thoracotomy. AIM OF THE STUDY: Indications and outcome of the surgery were assessed, based on the restrospective analysis data from subjects operated for blunt and penetrating thoracic injuries. SUBJECTS, METHODOLOGY AND RESULTS: 85 patients were operated for blunt or penetrating thoracic injuries during the period from 2000 to May, 2006. Urgent procedures for blunt thoracic injuries were performed in 13 subjects, and for penetrating injuries in 52 subjects. Immediate thoracotomy was indicated in 4 subjects suffering from stab injuries of the heart. 16 subjects were operated consequently for posttraumatic intrathoracic complications in blunt thoracic injuries. 4 subjects exited early postoperatively (4.7 %). CONCLUSION: The aim of urgent surgical interventions is to manage the major bleeding source (heart or large vascular injury), lungs, air leakage (tracheal or bronchial injuries) and diaphragmatic injuries. Irreversible hypovolemic shock in penetrating thoracic injuries and serious related injuries in blunt traumas resulted in peroperative and postoperative mortality. PMID- 17407948 TI - [Chylothorax. A case review and literature overview]. AB - Chylothorax is a rare disorder, predominantly of traumatic (iatrogenic) or tumorous aetiology. Its symptomatology complies with other types of fluidothorax. Long-term chylous depletion may result in malnutrition and other metabolic and immunological symptoms. Infectious complications are rare. The authors present a literature overview and their own case-review of chylothorax resulting from the suprarenal aortic aneurysm surgery. The conservative management failed and the complication in the 75-year old patient was managed using pleuroabrasion and talk pleurodesis. PMID- 17407949 TI - [Vascular surgery or endovascular treatment?]. PMID- 17407950 TI - [Autovenous ilicofemoral bypass, 21 years later --a case-review]. AB - We present the case of patient with autovenous ilicofemoral bypass after 21 years. The indication was traumatic demage of left iliac artery during car accident. The arterial reconstruction was patent for more than two decades with gradual arterialisation of graft. Excessive dilatation has caused the late occlusion of bypass. Ischaemia of the left limb was treated by crossover ilico femoral bypass. PMID- 17407951 TI - [Gastric carcinoid--a case review]. AB - Gastric carcinoid is a rare GIT endocrine tumor. The disorder is mostly asymptomatic, Clinically, it may present with pains, bleeding or obstruction. Onset of a carcinoid syndrom is a serious complication. The diagnosis of the disease is based on endoscopy and the tumor biopsy. Its therapy includes endoscopic or surgical resection. The resection type depends on the tumor size and its histological type. PMID- 17407952 TI - [Surgical-anatomical study as a part of operative treatment of primary hyperparathyroidism]. AB - Surgical- anatomical studies improve experience of a surgeon and technique of the parathyroids preparation. The authors conducted a study aimed at standardization of the parathyroidectomy procedure and collection of the parathyroids from cadaverous donors for allotransplantation. The total of 101 cadavers (51 females, 50 males) were sectioned. Parathyroid regions were closely preparated, the glands were collected and histologically verified. Anatomical differences of this region were recorded. The parathyroid tissue was confirmed microscopically in 75%. Mostly, the parathyroids were mistaken for lymphonodes, a thyroid or thymus tissue. Location of inferior parathyroids was more variable (28% of inferior parathyroids is located within the thymus tissue), compared to the superior ones. Based on the study, systematic anatomical preparation in the cervical region, including a. thyroidea inferior and n. laryngeus recurrens and around the thymus cervical procesi is recommended in parathyroid procedures, where preoperative location examinations were inconclusive. Complete thymectomy with exenteration of the anterior mediastinal adipose tissue is fundamental for preparations in the mediastinum. PMID- 17407954 TI - [Treatment of diaphyseal forearm fractures with locking intramedullary nail]. AB - INTRODUCTION: The aim of this study is to evaluate the results after treating diaphyseal fractures of the forearm with an inter-locking intramedullary nail. METHODS: Between 2003 and 2005, 45 patients with 68 fractures were treated using the ForeSight intramedullary nail (Smith&Nephew, Memphis, USA). The average patient age was 34.12 years. In the postoperative period, early and late complications were evaluated through radiological and functional methods. RESULTS: The average length of time to demonstrated bone healing was 13.2 weeks. 3 cases of prolonged healing were observed, and pseudoarthrosis formation did not occur. Assessment of function according to Anderson gave the following results: full range of movement in 86.9% of patients; mild restriction of movement in 10.9%; and severe restriction of movement in 2.2% of patients. Postoperative complications included: I superficial infection, 2 cases of incomplete radioulnar synostosis; and one case of compartment syndrome. CONCLUSIONS: Upon comparing the techniques of using plates with those of nailing in the treatment of forearm fractures, we have achieved comparable results with nailing, and we therefore regard it as advantageous foremost for treating open, serial and grossly comminuted fractures of the forearm bones. PMID- 17407953 TI - [The "inspection jejunostomy" after complex reconstructions of the central bile ducts]. AB - INTRODUCTION: Hutson and Russell described in 1984 a surgical technique, where a modified Roux-en-Y hepaticojejunostomy with the afferent limb brought up as jejunostomy after complex reconstructions of the central bile duct was performed [4]. It facilitates endoscopic retrograde access to the biliary tree for control examinations and biopsies in complicated reconstructive procedures after bile duct resection. We report our experience with this operative procedure. METHODS: After having performed complicated bile duct resection, the reconstruction was performed as a modified Roux-en-Y hepaticojejunostomy with the 20 cm afferent limb brought up as terminal jejunostomy in the right upper abdominal quadrant. Postoperative follow-up comprised endoscopic and radiologic controls of the biliary tree every 3 months for one year; ileostomy resection was performed one year later if the postoperative course was undisturbed. RESULTS: From 03/1995 to 07/2006 we performed this operation in 25 patients (mean age 62 yrs.). Indications were previous lesions of the common bile duct after laparoscopic (n = 10) or open cholecystectomy (n = 5), common bile duct resections in cholangiocellular carcinoma and gallbladder carcinoma with unclear intraoperative safety margins (n = 4), malignant granulosa cell tumour and simultaneous cholangiocellular carcinoma, focal nodal hyperplasia, Mirizzi-syndrome, cystadenoma of the pancreas head, cyst of ecchinococus granulosos and one patient with intrahepatic recurrent stone formation after orthotopic liver transplantation. The endoscopic and radiologic (cholangiography) diagnostic procedures--performed every 3 months postoperatively--were uneventful. CONCLUSIONS: The modified Roux-en-Y hepaticojejunostomy with the afferent limb brought up as jejunostomy permits good control and intervention possibilities in complicated bile duct surgery after bile duct lesions, tumor resection with unclear resectional margins and in recurrent intrahepatic stone formation. PMID- 17407955 TI - [Consequennce of nontreatment scapholunate instability of the wrist]. AB - Authors evaluate the results of patients with acute rupture of the scapholunate (SL) ligament, demonstrated by arthroscopy, in this study. One group of patients was treated with reinsertion of SL ligament by the help of Mitek anchor. In the other group there were patients with diagnosed total rupture of SL ligament which rejected the reinsertion of the ligament. Authors evaluate results of both groups 24 months after the injury. On the basis of clinical and X-ray results, even after such short time, reasonably worse results for the group without performed reinsertion were determined. Even X-ray documented arthrotic changes of scapholunate advanced collapse (SLAC) type were present. PMID- 17407956 TI - [Historical review of opinions on the nature of varicose veins and leg ulcers and their treatment]. AB - The development and changes of opinions concerning the nature and treatment od varicose veins and varicose ulcers are summarized in this article covering a long period from the Greek antiquity till the 20th century. Whereas in the ancient times (Hippokrates, Celsus, Galen) the meanings did not contradict the contemporaneous knowledge, curious opinions dominated from the 11th till the half of the 19th century, obviously influenced by the humoral theory of Avicenna. The official teaching claimed that black bile and other bad humours collected in varicose veins. Bandaging was thought to push back these humours into the body and cause madness or other serious illness and, therefore, was refused. On the contrary, ulcers were considered to be a favourable condition because their function was to drain the bad humours. The fundamentals of modern surgical treatment of varicose veins were presented by Madelung, Trendelenburg and Babcock at the end of the 19th and the beginning of the 20th century. Trendelenburg deserves acknoledgement of having introduced physiological and pathophysiological reasoning into the field of phlebology. The conception of the role of incompetent calf perforators for the development of leg ulcers advocated by Cockett and others represents a derailment on the way of evidence based medicine. The substance of this theory, although supported by most autors till the present time, have been repeatedly confuted by exact hemodynamic measurements. Evidence based medicine whose basement was laid by Trendelenburg more than 100 years ago cuts a difficult way through the field of phlebology. PMID- 17407957 TI - [Spontanneous intramural hematoma of the duodenojejunal junction mistaken for acute pancreatitis]. AB - Anticoagulation therapy may be complicated by spontaneous onset of an intramural hematoma of the duodenum and small intestine. Other causes were published only as case reviews. Radiological literature describes typical findings, which are fundamental for the diagnostics. However, only a fraction of patients present with them. This article describes a case of a female patient, treated for stomach pains. Based on her history, laboratory findings, ultrasound and CT findings, the condition was mistaken for acute pancreatitis and the pancreatic body necrosis with pseudocysts, drained under the CT control. The correct diagnosis was established a year later. Surgical revision indicated for a cystoid relaps and difficulties with intestinal passage revealed an old intramural hematoma within the duodenojejunal junction wall, the hematoma was evacuated and gastroenteroanastomosis performed. PMID- 17407958 TI - [Use of "Cypher" sirolimus-eluting stents in coronary heart disease patients with different types of coronary arterial lesions]. AB - The purpose of our study was to assess the immediate and late results of treatment with Cypher drug-eluting stents (Cordis, Johnson & Johnson, USA) in patients with coronary heart disease (CHD). This was a prospective study that included 738 patients who had been implanted Cypher stents in May 2002 to March 2006. The patients' mean age was 56 +/- 9 years; there were 87% of males. The patients were randomly included into the study and they underwent coronary stenting in the routine laboratory setting. A control group comprised 162 patients who had undergone Velocity or Sonic nondrug-eluting stents of the same firm, which had the similar structure. The groups did not differ in clinical characteristics. 827 stenoses in the eluting stent group and 225 stenoses in the control group were subject to revascularization. The immediate cure rate was 95 and 94%, respectively. The total number of events (myocardial infarction, emergency coronary bypass surgery, subacute occlusion of a stented segment) was 2.3% in the eluting stent group and 2.4% in the control group. A repeated examination 1 year after surgery was made in 482 and 119 patients in the drug eluting and nondrug-eluting groups, respectively. During the follow-up, one patient died of a extracardiac cause and 3 (0.6%) patients underwent coronary bypass surgery in the nondrug-eluting stent group; there were no deaths and 2 (1.6%) patients had coronary bypass surgery in the control group. In the eluting stent group, there were fewer cases of repeated endovascular procedures of target stenosis revascularization than in the control group (3.7% versus 11.7%; p < 0.0005). In the eluting stent group, the total number of unfavorable cardiovascular events was significantly less than that in the control group and it amounted to 3.3% as compared with 15.9% in the non-eluting stent group; p < 0.0005. Cardiovascular event-free survival was significantly higher in the eluting stent group: 92% versus 77% in the non-eluting stent group (p < 0.0005). PMID- 17407959 TI - [Comparative analysis of the X-ray morphology of coronary atherosclerosis and the results of endovascular interventions performed in the subacute period and 6 months after prior myocardial infarction]. AB - The study was undertaken to analyze the morphology of coronary lesion and the angiographic results of interventions made in the subacute period and 6 months after prior myocardial infarction. It included 593 patients with and without prior Q-wave myocardial infarction. The X-ray morphological parameters of the coronary bed were analyzed in the patients who had undergone invasive studies within a month (n = 362) and 6 months (n = 231) after sustained myocardial infarction. In both groups, the successful intervention rate was also estimated during endovascular treatment. The study showed that the number of stenoses with the complicated morphology (type B) was significantly more 6 months after sustained myocardial infarction than that in the subacute period. The number of occluded segments with the characteristics that were unfavorable for endovascular intervention (bridge collaterals, extended occlusions) was also significantly more in the late periods. In the subacute period, the angiographic success rate of endovascular recanalizations of chronic occlusions was 83.4%, which was significantly higher than that (62.6%) 6 months following infarction. The findings have led us to the conclusion that it is necessary to make routine coronary angiography in all postinfarction patients during their hospital stay in order to define the potentialities of early revascularization. PMID- 17407960 TI - [The comparison of glioblastoma and its surrounding edema by MRI of brain]. AB - In the article authors consider reasons, leading to the formation of edema around the glioblastoma by analysis of perifocal edema size, glioblastoma size and necrosis zone correlation. According their data, the size of glioblastoma edema, revealed by MRI, varies widely. Practically, in third of patients the size of edema is less than the glioblastoma size, in third--their sizes are equal, in 40% the size of edema exceeds the size of glioblastoma. Relying on received data it is found out that the formation of edema around the glioblastoma is not the consequence of tumor compression on adjoining brain tissue and is determined by transmissivity of blood-brain barrier disturbance as the result of vasoactive substances secretion by blastoma. PMID- 17407962 TI - [Differential magnetic resonance diagnosis of central lung cancer and acute pneumonia]. AB - The paper analyzes the authors' own data of chest magnetic resonance imaging (MRI) in 86 patients with verified central lung cancer and acute pneumonia. The MRI signs of lung cancer are systematized in exo-, endo-, and peribronchial forms of growth. The additional capacities of contrast enhancement are analyzed. The MRI semiotics of acute pneumonia has been developed. The differential diagnostic criteria for recognizing central lung cancer and acute pneumonia have been also elaborated. PMID- 17407961 TI - [The thyroid volume and methods determination]. AB - PURPOSE: to work-out method of the determination the volume of functioning thyroid tissue on the base of experimental investigations and to appreciate its accuracy. To analyze the clinical results of determination of the volume of the thyroid volume for different clinical groups with the use I123. MATERIAL AND METHODS: experimental investigations of the phantoms of different sizes were conducted, and also there were conducted the investigations of the phantoms with different diameters spheres which were inside the phantoms. On the base of conducted investigations there was suggested a method of the determination of the volume functioning thyroid tissue paying attention to the organ sizes and the microtexture of the picture. Summary error of this method with taking into consideration different factors was 7.02%. There were examined 36 patients with different kinds of thyroid diseases and patients of conditionally control group. Every patient was made planar method and SPECT with the use I123. RESULTS: the experimental investigations showed that for the determination of the volume of functioning tissue according SPECT data it should use the variable level of cut out of the background during analyzing the data paying attention to the organ sizes. The clinical investigations pointed out that there were trustworthy differences between signs indices among different clinical groups. Some signs are specifically for cancer, adenoma, Graves' disease. The most volume of nonfunctioning thyroid tissue is at cancer patients. CONCLUSION: there was worked out the method of determination of volume functioning thyroid tissue according to SPECT data. On the base of the received results there were offered integral indices which can be used for differential diagnose of the thyroid diseases. PMID- 17407963 TI - [Capacities of contrast enhancement magnetic resonance imaging versus X-ray and ultrasound studies in the detection of enlarged axillary lymph nodes in breast diseases]. AB - The authors conclude comparative data in the opportunity of detecting lymphadenopathy in patients with mammary gland pathology by X-ray ultrasound and MRI. High descriptiveness of MRI with contrast enchancement is marked in revealing lymphadenopathy of auxiliary lymph nodes. PMID- 17407964 TI - [Magnetic resonance imaging in the differential diagnosis of chronic endometritis: capacities and methodic features]. AB - By examining 83 females aged 17-48 years by magnetic resonance imaging (MRI), the authors conclude that the specificity and sensitivity of the technique without MR contrast agents in detecting chronic endometritis (CE) and chronic metroendometritis (CME) are 75.3 and 95.9%, respectively. On the MRI scans, hypertrophic and atrophic forms of CE have rather specific MR signs and appear as changes not only in the functional layer of the endometrium and transitional area, but also in the proper myometrium in CME. This all permits evaluating the degree of uterine wall involvement in the pathological process. Overall, the MRI criteria proposed by the authors can identify the signs of a chronic inflammatory process and its sequels and make a differential diagnosis this condition with female genital diseases to a high accuracy. PMID- 17407965 TI - [Local application of dimethyl sulfoxide at different concentrations to the prevention of radiation-induced damages in patient with cancer of the cervix uteri]. AB - OBJECTIVE: to study the efficacy of dimethyl sulfoxide ((DMSO) at different concentrations in preventing radiation-induced rectal and urinary bladder damages in patients with cervix uteri cancer (CUC). MATERIAL AND METHODS: combined radiation therapy (RT) was performed in 807 patients with CUC. In the control group (n = 221), RT was made, without applying radio-modified agents. An hour prior to a session of intracavitary irradiation, 10% DMSO solution was instilled into the rectum and urinary bladder in 113 patients and applications of metronidazole (MN) dissolved in 100% DSMO were made in 473 patients. Teleradiotherapy was performed, by using megavolt irradiation sources in the conventional fractionation mode; the total focal dose (TFD) was increased up to 40-46 Gy. Intracavitary irradiation was carried out on "AGAT-V" and "AGAT-VU" devices once weekly; the single focal dose in point A was 7 Gy; TFD was 49-56 Gy. RESULTS: 10% DMSO instillations reduced the incidence of late radiation-induced damages to the rectum and urinary bladder. In the control group, the incidence of these conditions was 19.0 and 9.5%, respectively; with the use of 10% DMSO, that was 8.8 and 7.1%. Applications of MN dissolved in 100% DMSO reduced the incidence of late radiation-induced damages to 1.7%. CONCLUSION: Local application of DMSO is a method for preventing late radiation-induced damages to the rectum and urinary bladder in patients with CUC. When the concentration of DMSO is increased, its preventive effect increases. PMID- 17407966 TI - [Solid pseudopapillary tumors of the pancreas: two cases]. PMID- 17407967 TI - [Chondromyxoid fibroma of the heel bone]. PMID- 17407968 TI - Milk-derived proteins and peptides of potential therapeutic and nutritive value. AB - Milk and colostrum are rich in proteins and peptides which play a crucial role in development of the immune system in mammalian offspring. Immunotropic properties of these compounds prompted investigators to search for their utility in prevention and therapy of various disorders in humans. The following constituents of milk are of particular interest: 1) Lactoferrin (LF)--exhibits antibacterial, antifungal, antiviral, antiparasite and antitumor activities. It is protective with regard to intestinal epithelium, promotes bone growth and accelerates recovery of the immune system function in immunocompromised animal; 2) A Proline Rich Polypeptide (PRP) shows a variety of immunotropic functions, including promotion of T-cell maturation and inhibition'of autoimmune disorders. PRP was recently found to improve or stabilize the Instrumental Activity of Daily Living status in Alzheimer's disease patients. 3) Casein--has been protective in experimental bacteremia by eliciting myelopoiesis. Casein hydrolyzates were also protective in diabetic animals, reduced the tumor growth and diminished colicky symptoms in infants. Casein-derived peptides have been found to have antihypertensive effects. Glycomacropeptide (GMP)--a peptide derived from kappa casein, exhibits antibacterial and antithrombotic activities. 4) Alpha lactalbumin (LA)--demonstrates antiviral, antitumor and anti-stress properties. LA-enriched diets were anxiolytic, lowered blood pressure in rats, prevented diarrhea and led to a better weight gain in malnourished children. 5) Lysozyme- is effective in treatment of periodentitis and prevention of tooth decay. Milk enriched in lysozyme was used in feeding premature infants suffering from concomitant diseases. 6) Lactoperoxidase--shows antibacterial properties. In conclusion, milk-derived proteins and peptides are bio-accessible and safe for the prevention and treatment of numerous disorders in humans. PMID- 17407969 TI - Antitumor and immunomodulatory effect of coumarin and 7-hydroxycoumarin against Sarcoma 180 in mice. AB - The antitumor effect of peroral treatment with coumarin and its main metabolite in humans 7-hydroxycoumarin (7-OHC) against Sarcoma 180 in mice was studied. Both agents inhibited tumor growth and increased survival time of tumor-bearing animals. The antitumor effect was better when coumarins were administered prior to tumor inoculation suggesting that the immunomodulatory potential of coumarins might exceed their well-known direct cytostatic activity. The additive effect of coumarins in combination with a suboptimal LPS dose in tumor growth inhibition was demonstrated. Coumarin treatment enhanced the macrophage migration activity in the presence and absence of LPS and increased nitric oxide release. In vitro, coumarins induced IL-12 in murine macrophages and additively increased the LPS induced IL-12 release. These data indicate that the immunomodulatory activity of coumarins contributes to their direct cytostatic effect and demonstrate their potential to combine as immunostimulators with other antitumor agents. PMID- 17407970 TI - Dip1 inhibits growth and gene transcription in MCF-7 breast cancer cells. AB - In previous studies we identified a novel gene Dipl, also designated CCNDBP1, which encodes a 42kDa helix-loop-helix (HLH) nuclear protein. Although this protein was originally identified by its ability to bind to cyclin D1 its precise biochemical functions are not known. In the present study we carried out mechanistic studies on Dip1 focusing on the human breast cancer cell line MCF-7. We found that overexpression of Dip1 in MCF-7 cells inhibited colony formation and cell proliferation. Reporter assays in MCF-7 cells indicated that Dip1 strongly inhibited the transcriptional activities of the cyclin D1, c-fos, NF kappaB, SRE and p21cP1 promoters. Furthermore studies with truncated and mutant forms of the cyclin D1 promoter suggest that Dip1 does not act on specific transcriptional elements. Assays with mutant and truncated forms of Dip1 indicated that both the LXXLL motif and the HLH domain play important, but not exclusive, roles in these inhibitory effects. Dip1 co-immunoprecipitated with the histone deacetylase (HDAC) proteins HDAC1 and HDAC3. Nevertheless Dip1 markedly inhibited the stimulation of cyclin D1 promoter activity obtained with trichostatin A [1], an inhibitor of HDAC. Taken together these findings suggest that Dip1 functions as a general repressor of transcription. Although the precise mechanism by which Dip1 inhibits gene transcription and the growth of MCF-7 cells remain to be determined, the present results suggest that Dip1 is a candidate tumor suppressor gene. PMID- 17407971 TI - A nanoemulsion of an anti-oxidant synergy formulation reduces tumor growth rate in neuroblastoma-bearing nude mice. AB - Neuroblastoma, the most common form of childhood cancer, may arise from a biochemical block of cellular differentiation and a resultant continuation of a proliferative state. Neuroblastoma often spontaneously reverts by undergoing partial differentiation and ultimate degeneration and may be associated with the generation of reactive oxygen species (ROS). We have recently reported in neuroblastoma cell culture studies that an anti-oxidant synergy formulation (ASF) can induce differentiation and buffer neuronal degeneration and oxidative stress in cultured cortical neurons and in central nervous system tissue of apolipoprotein E-deficient mice. The objective of the present study was to investigate whether a subcutaneous injection and/or transdermal application of a nanoemulsion preparation of ASF would reduce tumor growth rate in a neuroblastoma xenograph mouse model. The results indicate that whereas suspensions of ASF were ineffective in decreasing tumor growth rate in the neuroblastoma mouse model, tumor growth rate was similarly reduced an average 65% by either subcutaneous injection or transdermal application of an ASF nanoemulsion preparation to the tumor. In conclusion, the data suggest that subcutaneous and/or transdermal application of an ASF nanoemulsion preparation is effective in reducing tumor growth rate in this neuroblastoma mouse model. PMID- 17407972 TI - Differential protein expression patterns obtained by mass spectrometry can aid in the diagnosis of Hodgkin's disease. AB - More than 90% of patients with cancer, if diagnosed early, can be promptly treated; however diagnosis usually occurs after cancer cells have metastasized. Recent technological advances in mass spectrometry challenges the field of machine learning to model such high dimensional datasets for clinical diagnosis and prognosis. Here we use support vector machines recursive feature elimination to hunt for protein expression patterns in the serum mass spectra of Hodgkin's disease (HD) patients and control subjects (CS) that could aid in diagnosing-the disease. Based on eight selected features, support vector machines was able to correctly classify among all CS and HD patients based on the leave-one-out. We also correctly classified an independent dataset, acquired from the same samples, with the previously generated SVM model. PMID- 17407973 TI - American ginseng (Panax quinquefolius L.) extract alters mitogen-activated protein kinase cell signaling and inhibits proliferation of MCF-7 cells. AB - Ginseng has been shown to inhibit cancer cell proliferation and tumor growth, however the mechanisms underlying this inhibition have yet to be elucidated. An inhibitory effect of hot water-extracted American ginseng (Panax quinquefolius L.) root on cell proliferation was demonstrated using MCF-7 human breast cancer cells treated with a wide concentration range of the ginseng extract (GE) for 6 days. The effects of GE were concentration-dependent with an IC50 of 0.49 microg/microl and the minimum exposure time to elicit an inhibitory response was 24 hours. Using an antibody microarray, it was determined that several key cell survival proteins were altered in GE-treated cells, including several members of the mitogen-activated protein kinase (MAPK) family. A GE-induced decrease in phospho-MEK1/2 and -ERK1/2 and an increase in phospho-Raf-1 were observed and verified using Western blot analysis. Furthermore, mRNA and protein expression of the Raf-1 kinase inhibitor protein (RKIP) was shown to be transiently, yet significantly, upregulated following GE treatment. These results suggest that American ginseng may act to inhibit breast cancer cell proliferation by increasing the expression of RKIP, resulting in inhibition of the MAPK pathway. This novel mechanism has implications in the potential prevention and treatment of breast cancer. PMID- 17407974 TI - Somatic tissues degeneration. PMID- 17407975 TI - Spontaneous unusual expression of frequency of chromosome aberrations and common fragile in human lymphocytes of colorectal cancer patients induced by Aphidicolin. AB - Chromosomal fragile sites are distributed all over the human genome. Aphidicolin mediated expression frequency of common fragile sites and other chromosomal changes were evaluated in prometaphase/metaphase chromosomes obtained from peripheral blood lymphocytes of colorectal cancer patients. The present study reveals first time high incidence i.e. 6 % of aphidicolin induced chromosome breaks / gaps designated as "common fragile sites" in cell population of clinically diagnosed patients of colorectal cancer patients in Nepalese population. These chromosomal changes including structural and numerical were compare to clinically healthy normal individual of same sex / age groups, act as controls for statistical analysis. The frequency of chromosomal aberration in cancer patients were significantly higher (p<0.001) when compare to normal individuals. The increased genetics instability probably either due to nutritional factor i.e. lack of folic acid component in diet--an essential component required for DNA synthesis or unknown environmental factor for such genetic disorder. The present study indicates aphidicolin high frequency of induced chromosome aberrations and "common fragile sites" because of late replication of DNA in mitosis in colorectal cancer patients suggesting these sites could be used as suitable marker for determining genetic predisposition in cancer patients. PMID- 17407976 TI - Outcomes of dead on arrival children: is it time to teach everybody PBLS? PMID- 17407977 TI - Cancers in infancy: percent distribution and incidence rates. AB - Cancer occurring in infants often has clinical and biological properties that are different from those of the same histologic type of cancer occurring in older children. The histologic distribution of cancers in infants and that in older children are also different. We collected infant cases diagnosed as having cancer from the database of the Cancer Registry in our Medical Center between 1995 and 2004. One hundred and twenty infants (66 males and 54 females), including 17 neonates, were diagnosed with cancer over this 10-year period. The top five cancer subtypes were: leukemia in 34 infants (28.3%), retinoblastoma in 19 (15.8%), neuroblastoma in 16 (13.3%), germ cell tumor in 15 (12.5%), and brain tumor in 14 (11.7%). The overall disease-free survival rate was 54. 2% (65/120) with a median follow-up duration of 7.6 years for the survivors. From the cancer registry in our hospital during the same period, 1995-2004, infant cases accounted for 9.5% of the total 1,269 children with cancer diagnosed at the age younger than 15 years. The percent distribution of the major types of cancer was different in the infant and the childhood group. According to the Taiwan Pediatric Oncology Group's data analysis, the infant cancer incidence rate in Taiwan is 207.6 per million infants for years 1995-2004. Our study confirmed that the prognosis is very poor for infant leukemia and rhabdoid tumor, while that is good for embryonal tumors and germ cell tumors occurring in the infancy. PMID- 17407978 TI - Predictors of successful resuscitation in non-trauma dead-on-arrival children. AB - BACKGROUND: Dead-on-arrival (DOA) children pose difficult challenges in resuscitation because of the very low survival rate. In this study, we aimed to analyze the factors that may be related to predicting successful cardiopulmonary resuscitation (CPR). METHODS: We reviewed the hospital records of 120 DOA children aged younger than 18 years who had been admitted to the emergency department (ED) from 2000 to 2004 and analyzed related factors that may have influenced initial CPR in the non-trauma DOA children. Survival analysis was used to compare differences in survival rate between the non-trauma and trauma DOA children. Receiver operating characteristic (ROC) analysis was used to determine the predictive in-hospital CPR duration related to success of initial CPR. RESULTS: We found the initial cardiac rhythm (P = 0.007), pre-hospital basic life support (BLS) (P < 0.001), mode of transportation (P = 0.019), the period from scene to hospital (P=0.025) and the duration of pre-hospital BLS (P = 0.003) were the significant factors related to initial successful CPR in non-trauma DOA children. Based on the ROC analysis, the cutoff value of in-hospital CPR duration was 23 minutes in non-trauma DOA children. CONCLUSIONS: We found that in-hospital CPR should be performed for at least 23 minutes in nontrauma DOA children for spontaneous circulation to return. PMID- 17407979 TI - Factors affecting the non-compliance for follow-up in very low birth weight children. AB - BACKGROUND: Long-term longitudinal follow up of very low birth weight (VLBW, birth weight < or =1500 g) infants is important. This study assessed the factors affecting non-compliance for follow-up at different ages in VLBW children. METHODS: The demographic data, morbidity and neurodevelopmental data of 321 VLBW infants were analyzed between those returned and not returned. RESULTS: The majority of the children who did not attend the follow-up clinic were caused by inaccessibility, and refusal by the parents. The non-compliant children had higher rate of multiple births, short paternal education years, mother being a housewife, and lower incidence of first child, emergent caesarian section and small for gestational age. They did not differ much from the children returned in the occurrence of neonatal diseases. No significant difference of the growth and neurodevelopmental status at the previous visit could be observed between children of the two groups. When data of different follow-up visits were compared, it was noted that the most common reason for not attending the follow up visits was inaccessibility for the 6- and 12-month visits and refusal by the parents for other visits. Multivariate logistic regression analysis revealed that variable(s) increasing the non-compliance were none for the 6-month visit, severe retinopathy of prematurity for the 12-month visit, short duration of paternal education for the 18-, 24- and 36-month visits and multiple births for the 36 month visit. CONCLUSIONS: These results showed that the reasons and the affecting variables for not attending the follow-up assessment were different for different age groups. PMID- 17407980 TI - Penile length of normal boys in Taiwan. AB - OBJECTIVE: Micropenis is usually defined as a penis that has a stretched length less than 2.5 standard deviations below the mean value. The purpose of this study was to establish a database and set referable standards of penile length for Taiwanese boys. METHODS: A total number of 2126 boys (156 male newborns, 1198 male infants under 2 years old, and 772 boys older than 2 years old) were included in this study. We excluded those boys with congenital anomaly, frankly genital anomaly and congenital heart disease. Both stretched and flaccid penile lengths were measured for comparison. RESULTS: Our data revealed that the average penile length increased with chronologic age (about 3 cm in neonates, 4 cm when 1 year old, and near 5 cm when 5 years old). But it also revealed that Taiwanese boys have slightly shorter stretched penile length after newborn period till 5 years old when compared with Caucasian boys (all P values < 0.05). CONCLUSIONS: Normal stretched penile length varied between different ethic groups, and maybe body size contribute more or less to smaller penile size in Taiwanese boys compared to Caucasian boys in light of general knowledge that Caucasian is taller than Chinese (including Taiwanese) in average. PMID- 17407982 TI - Clinical manifestations of chromosome 21 interstitial deletion: report of four cases. AB - Chromosome 21 is the smallest human chromosome; it contains 1.7% of the human genome, or about 54,000 kb. Multiple abnormalities have been observed in patients with a deletion of the long arm of chromosome 21. Phenotypic variability between cases involving different deletions of 21q have been described. We report four cases of deletion (21)(q11.2-q22.1). Three of the reported cases are familial, involving a mother and her two sons, and one is de novo. All cases had mild mental retardation, antimongolian slants, long philtrum, and protruding tongue as well as open mouth. Hypertonia was noted when each was young. Arachnoid cyst, cerebral dysfunction, thalassemia minor, and growth hormone deficiency were found in the de novo case. This is the first report of chromosomal deletion (21)(q11.2 q22.1) in the Taiwanese population. PMID- 17407981 TI - Juvenile recurrent parotitis. AB - BACKGROUND: Juvenile recurrent parotitis (JRP) is a rare, recurrent non obstructive, nonsuppurative parotid inflammation in young children with a multifactorial etiology. METHODS: The records of 12 children with recurrent parotitis were retrospectively reviewed. RESULTS: The age of onset ranged from 3 to 8 years (mean, 5.1 years). Each attack lasted an average of 3.5 days. The major clinical manifestations included fever (75%), swelling (100%), and pain (100%). One girl also had xerostomia and keratoconjunctivitis sicca. Her biopsy specimen from a minor salivary gland was consistent with juvenile Sjiigren's syndrome. Six patients had positive antinuclear antibodies (4 with a speckled pattern and 2 each with a homogeneous or nucleolar pattern). Bilateral sialography was performed in 9 children. The results in 4 were normal, 2 had unilateral punctate sialectasis, and 3 had bilateral sialectasis. After sialography, the frequency of recurrences significantly decreased from 5.11 to 0.56 per year (P < 0.05). CONCLUSIONS: Bilateral sialography is useful for the diagnosis of JRP, but it also appears to decrease the frequency of recurrences. PMID- 17407983 TI - Holocarboxylase synthetase deficiency: report of one case. AB - Holocarboxylase synthetase (HCS) is an enzyme that catalyzes biotin incorporation into carboxylases, and its deficiency causes biotin-responsive multiple carboxylase deficiency. We report a patient who had his first episode at 32 months of age. The main clinical findings were a characteristic rash, projectile vomiting, progressive consciousness loss, organophosphate order, and hypotension. Laboratory examinations showed metabolic acidosis with ketolactic acidosis, hyperammonemia, and urine organic acid profile suggestive of a biotin utilization abnormality consistent with multiple carboxylase deficiency. Nucleotide sequence analysis of the biotinidase gene of the patient revealed negative finding, however, analysis of HCS gene found a homozygous 1809C->T (R508W) mutation. R508W is a rare mutation in Taiwanese HCS deficiency patients, which is associated with the late-onset phenotype. The patient responded dramatically to biotin, and has remained normal growth and development during more than three years of follow-up. Therefore, a high index of suspicion for timely diagnosis and treatment could prevent severe complications. PMID- 17407984 TI - Short stature in patients with 45,X/46,XY mosaicism: report of three cases. AB - Chromosomal analysis is routinely considered in all girls--but not boys-of short stature to screen for Turner syndrome. We report three patients of short stature (body height < 3rd percentile) with 45,X/46,XY mosaicism karyotype. They were brought to our outpatient department at the ages of 10, 13, and 18 years, respectively. Two of them had some stigmata of Turner syndrome. Two were treated with growth hormone. In the first patient, body height increased by 20 cm in 3 years 7 months. The second case increased by 6.5 cm in 8 months. We suggest that boys with unexplained short stature should be screened with chromosomal analysis, and growth hormone treatment should be offered to boys of short stature and 45,X/46,XY mosaicism before the epiphyseal plates close. PMID- 17407985 TI - [Intestinal occlusion in a patient bearing a ventriculo-peritoneal shunt]. PMID- 17407986 TI - [To improve the quality of the studies is to improve the quality of the journal]. PMID- 17407987 TI - Indomethacin reduces short-circuit current and oxygen consumption in normal and chronically hypoxic rat colon. AB - Chronic hypobaric hypoxia is a physiological environmental stressor. While its effects on most major organ systems have been extensively studied, few works have addressed hypoxia-induced changes in intestinal transport. The effects of cyclooxygenase blockade with indomethacin on short-circuit current (Isc) and oxygen consumption (QO2) of the distal colonic epithelium of control rats and rats submitted to hypoxia for 10 days at 0.52 atm were studied. Isolated mucosae were mounted in an Ussing chamber modified for measuring QO2 while preserving transepithelial vectorial transport. Amiloride was added to the mucosal hemichamber to block a sodium component of Isc present in hypoxic rats. In this condition, basal Isc did not differ between the hypoxic and the control group, but QO2 was higher in the former. Indomethacin (30 micromol/L) reduced Isc to the same extent in both groups, but QO2 reduction was larger in the hypoxic group. Pharmacological blockade of chloride secretion and a low-chloride solution abolished the indomethacin-induced reductions of Isc in both groups, and the reduction of QO2 in controls, and attenuated but did not suppress the QO2 reduction in the hypoxic group. Linear regression analysis of QO2 changes versus Isc changes yielded a significant correlation for both groups, with regression lines with the same slope, but a higher position in bypoxic animals. Results suggest that spontaneously releasedprostaglandins are equally important for maintaining colonic chloride secretion in hypoxic as in normoxic rats, but that, in the former, indomethacin has an additional effect on QO2 which is unrelated to ion transport. PMID- 17407988 TI - [Molecular characterization of hepatitis E virus in three acute liver failure cases in children in Argentina]. AB - Strains of hepatitis E virus (HEV) isolated from Argentinian patients with sporadic hepatitis, as well as from swine from Argentina, belong to genotype 3. HEV genotype 3 variants have been described associated with acute liver failure (ALF) in adults from Japan and the United Kingdom. In Argentina, 30% of ALF in adults and children are of unknown aetiology. To study if HEV could be an aetiological agent associated with ALF in children, serum and/or fecal samples fJom 35 children (mean age: 6 years, 20 female, 15 male) were analyzed during 2003 and 2004. HEV RNA was detected by RT-nested PCR with primers designed within ORF 1 and ORF 2 regions. HEV RNA could be detected in three cases. Two were 12 year-old boys fom Buenos Aires province and the third was a 3-year-old girl from Corrientes province. Sequence analysis indicates that the three isolates are distinct from each other but all belong to genotype 3, exhibiting a close relationship with swine and human strains fJom sporadic cases of HEV, previously reported in Argentina. This data suggests a potential link between ALF and HEVin children in Argentina and indicates the need for the determination of HEV status in the differential diagnosis in ALE Further studies would aid in determining the true impact of this infection in Argentina and the potential benefits of a vaccine against HEV presently in phase III trials. PMID- 17407989 TI - [Liver evaluation with multidetector CT. Angiotomography, volume determination and virtual hepatectomy]. AB - AIM: to show a new technique of presurgical liver tumor evaluation using multidetector computed tomography (MDCT), determining the usefulness of an giographic reconstructions, presurgical virtual hepatectomy and 3D liver volume determination, in correlation with surgery findings. METHODS: twenty patients with primary or secondary liver tumors were evaluated with MDCT and then operated on. Dual-phase CT was performed in all patients on a 4-row multidetector CT scanner (Mx8000; Philips Medical Systems) after mechanical injection of 120 ml of iodinated contrast medium. Scanning was performed using a detector configuration of 4 x 2.5 mm. Images were sent to a workstation and they were analysed with the surgeons. The 3D volumes of each lesion, of the total liver and of the segments to be resected were calculated. Vascular reconstructions and virtual hepatectomy were also performed. Correlation of the liver volume between MDCT and surgery was calculated using the Bland and Altman method. RESULTS: virtual liver segmentation allowed to perform the surgery in 100% of the patients in one time and there were no complications. The correlation coefficient was 0.83 (CI 95%: -132.08, 159.78). CONCLUSIONS: presurgical liver hepatectomy is a new application tool of MDCT The angiographic findings and the volume determination are useful to determine the surgical techniquefor each patient and this information allows the surgeons to know if the remnant liver will be enough for the patients to avoid a post surgical liver insufficiency. PMID- 17407990 TI - Decrease in prevalence of peptic ulcer and gastric adenocarcinoma at the Policlinico Peruano Japones, Lima, Peru, between the years 1985 and 2002. Analysis of 31,446 patients. AB - AIM: To determine the prevalence of peptic ulcer and gastric adenocarcinoma in a population of middle and high socio-economic status in Lima, Peru, between 1985 and 2002. MATERIAL AND METHODS: The prevalence of gastroduodenal ulcer and gastric adenocarcinoma was determined after studying by esophagogastroduodenoscopy 31,446 patients at the Policlinico Peruano Japonds (PPJ) between 1985 and 2002. All patients had chronic upper gastrointestinal symptoms and had endoscopically proven diagnosis of active peptic ulcer or histological diagnosis of gastric adenocarcinoma. RESULTS: Analysis of PPJ population identified a decrease in the prevalence of gastric ulcer and duodenal ulcer from 3.15% and 5.05% respectively in 1985, to 1.62% and 2.00% respectively in 2002. Gastric adenocarcinoma prevalence also decreased from 3.19% in 1988 to 0.92% in 2002. The prevalences of gastric ulcer (OR = 1.49, 95% IC 1.26-1.77, p < 0.001), duodenal ulcer (OR = 1.32, 95% IC 1.15-1.51, p < 0.001) and gastric adenocarcinoma (OR = 1.53, 95% IC 1.25-1.86, p < 0.001), decreased in the last eight years. CONCLUSIONS: Over the study period, there was a significant decrease in the prevalence of peptic ulcer and gastric adenocarcinoma at the Policlinico Peruano Japones, where patients from middle and high socio economic status are attended. PMID- 17407991 TI - [Abdominal bleeding due to spontaneous mesenteric vein rupture]. AB - Hepatic cirrhosis is the leading cause of portal hypertension and is usually associated with the development ofsplacnic varices. Variceal intraabdominal rupture is a rare cause of hemoperitoneum. A case of spontaneous mesenteric vein rupture is reported. CLINICAL CASE: 43-year-old man with hepatic cirrhosis (Child Pugh C stage) and previous bleeding of esophageal varices, admitted to hospital because of orthostatic hypotension episodes (positive tilt test) and abdominal distention. Laboratory: anemia, low platelet count and abnormal coagulation tests. CT scan showed abdominal fluid. Exploratory laparotomy was performed and 3 liters of blood were found into the abdominal cavity. The mesenteric vein had a ruptured variceal dilatation with intermitent jet bleeding. No organ abnormalities were found. Repair of the dilated vein was performed. The patient evolved well and was dismissed from hospital ten days after admittance. OBJECTIVE: To present a case in which a rare cause of intraabdominal bleeding, usually associated with high mortality rate, is follawed by a good clinical evolution. PMID- 17407992 TI - [New evidence in the management of digestive bleeding due to duodenal ulcer]. PMID- 17407993 TI - Access to and satisfaction with care comparing patients with and without serious mental illness. AB - OBJECTIVES: We compared perceived access to and satisfaction with health care between patients diagnosed with serious mental illness (SMI: schizophrenia or bipolar disorder) and among those with no SMI diagnosis. METHOD: We conducted a national, cross-sectional study of VA patients in Fiscal Year (FY) 1999 (N = 7,187) who completed the VA's Large Health Survey of Veteran Enrollees (LHSV) section on access and satisfaction and either received a diagnosis of schizophrenia or bipolar disorder, or did not and were randomly selected from the general non-SMI VA patient population (non-SMI group). We compared the probability of perceived poor access and dissatisfaction using multivariable logistic regression adjusting for patient covariates. RESULTS: Compared to non SMI patients, patients diagnosed with bipolar disorder were more likely to report difficulty in receiving care they needed (adjusted OR = 1.36,p < .05) or seeing a specialist (adjusted OR = 1.44, p < .001). Patients diagnosed with schizophrenia were more likely to report dissatisfaction, including thoroughness by their provider (adjusted OR = 1.37, p < .001) and the provider's explanation of problems (adjusted OR = 1.54, p < .001) compared to non-SMI patients. CONCLUSIONS: Patients diagnosed with bipolar disorder reported greater problems with access to health care, while those diagnosed with schizophrenia were less satisfied with the process of care. PMID- 17407994 TI - The accurate diagnosis of early-onset dementia. AB - Early-onset dementia (EOD, < 65 years at onset) is a relatively common and frequently misdiagnosed condition. One reason for misdiagnosis is that EOD has a more varied differential diagnosis than late-onset dementia (LOD). For example, Alzheimer's disease (AD), the preponderant LOD, makes up only about one-third of EODs; the rest are due to vascular dementias, frontotemporal lobar degenerations, traumatic head injury, alcohol-related dementia, and a great many other conditions. Another reason for misdiagnosis is that early-onset AD may have predominant cognitive deficits other than memory loss and a potential familial inheritance with spastic paraparesis, seizures, or myoclonus. A third reason is that EOD often presents with neuropsychiatric features out-of-proportion to any cognitive deficits. Despite these obstacles, it is important to accurately diagnose EODs, particularly because they differ in management and course. Clinicians can successfully diagnose most EODs with careful cognitive and family histories, mental status and neurological examinations, and neuroimaging. PMID- 17407995 TI - Depression and smoking across 25 years of the Normative Aging Study. AB - OBJECTIVE: The majority of past findings indicate that smokers are more likely than non-smokers to report depressive symptoms and that depression may act as an impediment to smoking cessation. The aim of the present study is to examine the stability of the relationship between depressive symptoms and smoking status and to determine whether the presence of depressive symptoms predicts continued smoking. METHODS: Subjects were initially healthy men (n = 2208) from the Veterans Administration Normative Aging Study, an ongoing cohort of older men who have been re-assessed every 3-5 years for a period of 25 years. Depressive symptoms measures employed were the Cornell Medical Index, the MMPI-2 Content Depression Scale, and the Center for Epidemiological Studies Depression Scale. RESULTS: Depression scores were higher among continuing smokers compared to never and former smokers and those who quit after entering the study. None of the three depressive symptoms measures were associated with decreased likelihood of smoking cessation. The change in MMPI-2 depression scores observed in a 4-year follow-up was the same among those who quit and those who remained smokers. CONCLUSIONS: Higher prevalence of depressive symptoms among male smokers is stable across time and is likely to contribute to higher morbidity and mortality among these smokers. However, presence of depressive symptoms did not have a significant impact on smoking cessation. PMID- 17407996 TI - Occult PTSD with panic attacks in a patient post-TIA: case report. AB - Affective disorders are common sequelae of cerebrovascular events. A myriad of evidence demonstrates that clinically significant depression can often follow a stroke. However, less is known about the extent to which anxiety disorders present after these experiences, and in particular, post-traumatic stress disorder (PTSD) with panic attacks. To our knowledge, the association between transient ischemic attacks (TIAs) and PTSD with panic attacks has not been described in the literature. Here we describe a patient with depression and PTSD with panic attacks following a transient ischemic attack. Many non-psychiatric physicians learn about post-stroke depression so they are equipped to screen for it. However, to many physicians, PTSD with panic attacks usually conjures the image of an individual who experienced either trauma or rape. Therefore, it is important to increase the awareness of such complications after TIAs. Increased awareness of these TIA sequelae has important implications for detection by non psychiatric healthcare providers. Furthermore, prompt recognition and treatment has important implications for patient quality of life. PMID- 17407997 TI - Frequency of attendance at religious services, cardiovascular disease, metabolic risk factors and dietary intake in Americans: an age-stratified exploratory analysis. AB - BACKGROUND: Few data have been published on the association of attendance at religious services with cardiovascular morbidity and dietary and metabolic risk factors in representative samples of populations despite a known inverse association with mortality and smoking. OBJECTIVE: To test the null hypothesis that frequency of attendance at religious services is unrelated to prevalence or levels of cardiovascular disease, dietary and metabolic risk factors. DESIGN: Cross-sectional survey of a large national sample. PARTICIPANTS: American men and women aged 20 years and over with complete data in the Third National Health and Nutrition Examination Survey (N = 14,192). MEASUREMENTS: Self-reported frequency of attendance at religious services, history of doctor-diagnosed diseases, food intake frequency, 24-hour dietary intake, health status, socio-demographic variables and measured serum lipids and body mass index. RESULTS: Weekly attenders were significantly less likely to report stroke, even after adjusting for multiple variables only in African American women OR = 0.35, 95% CI 0.19 0.66, p < 0.01. No association was seen for heart attack or diabetes. Fish intake at least weekly was more common in weekly attenders, significantly so only in African American women (odds ratio 1.24, 95% CI 1.01-1.58, p < 0.05) and in older Mexican American men (odds ratio 2.57, 95% CI 1.45-2.57, p < 0.01). In linear regression analyses, no significant independent associations were seen between attendance frequency and serum lipid levels or dietary intake of energy, or fat in g and % of kcal. CONCLUSION: Hypotheses generated by these analyses are that in African American women stroke is less prevalent and weekly fish intake more prevalent among weekly attenders than others and that there are no significant independent associations of serum lipids, dietary intake, prevalent CHD, or diabetes with frequency of attendance of religious services. Independent testing of these hypotheses in other samples is needed. PMID- 17407998 TI - Case report of 3 patients with severe mental illness and chronic hepatitis C virus infection treated with interferon-alpha. AB - We report on three patients with hepatitis C virus infection, severe mental illness, and substance use disorders that we treated successfully with interferon and ribavirin. This population has historically been refused such treatment on the grounds that they were unlikely to be adherent to treatment or would experience untoward psychiatric side effects. These case reports add support to the growing body of evidence that persons with severe mental illness and substance use disorders can be successfully treated for hepatitis C with careful monitoring and psychiatric oversight. PMID- 17407999 TI - A cohort study to assess the impact of depression on patients with kidney disease. AB - AIMS: This study assessed the impact of depression on mortality and quality of life of patients with kidney disease. MATERIAL: This study was conducted at Hospital Sao Lucas of Pontificia Universidade Catolica of Rio Grande do Sul, Porto Alegre, Brazil, with a cohort of 125 patients seen from 1996 to 2004. (mean follow-up = 77.6 months). Patients undergoing renal replacement therapy were asked to fill in the Beck Depression Inventory (BDI). At the completion of follow up, 72 patients were alive, and 56 answered the BDI and the Medical Outcome Study Short Form General Health Survey (SF-36) to assess quality of life. Main outcome measures were mortality and quality of life. RESULTS: Initial analysis showed that mortality was associated with age, treatment modality--continuous ambulatory peritoneal dialysis (CAPD) or hemodialysis (hd)--and depression. When controlling for age, gender, depression, and treatment modality in a multiple regression analysis, transplantation (tx), gender and age was associated with lower mortality. However, levels of depression at the end of follow-up were strongly associated with quality of life. CONCLUSIONS: Depression was not associated with mortality of patients with kidney disease when controlling for gender, age, and treatment modality. Transplantation was the main factor associated with lower mortality. Depression was, however, a strong predictor of quality of life. Number of depressive symptoms was directly associated with lower quality of life in most of the items of the SF-36. PMID- 17408000 TI - Quality of life and associated factors among adults with epilepsy in Nigeria. AB - OBJECTIVE: Epilepsy is a common condition worldwide and has been observed to affect quality of life (QOL). Though, much has been written on this subject among western populations, little research has been done in developing countries of Africa including Nigeria. The study aims to identify factors associated with quality of life among adult epilepsy patients in this environment. METHOD: Respondents were evaluated using the 10-item Quality of Life in Epilepsy Inventory (QOLIE-10), the 30-item General Health Questionnaire (GHQ-30), the modified Mini Mental State Examination (mMMSE),and the Hospital Anxiety and Depression Scale (HADS). Diagnosis of epilepsy was based on clinical and electroencephalographic findings. RESULTS: The mean age of the 51 respondents was 27.7 years (SD = 9.7). Thirteen (25.5%) had an average of 2 seizure episodes in the month preceding the interview, while 37 (72.5%) have had the condition for more than 5 years. Factors that were significantly associated with overall quality of life included being female (p < 0.05), seizure frequency (p < 0.01), using more than 1 anti-epileptic drugs (AEDs) (p < 0.01), GHQ-30 score (p < 0.01), high anxiety score (p < 0.001), and high depression score (p < 0.01). Multiple regression analysis showed that depressive symptoms were the single most important factor explaining low QOL. Other factors were GHQ-30 score, seizure frequency, and being a woman. CONCLUSION: Controlling seizures and paying attention to the psychological needs of adult epileptics will have a positive effect on the QOL among Nigerian epileptics. PMID- 17408001 TI - Correlates of quality of life in primary care patients with hypertension. AB - BACKGROUND: Hypertension has been associated with lower levels of quality of life (QoL). However, the specific correlates of lower QoL in this patient population have remained largely unclear. METHODS: A cross-sectional design of 1,125 primary care patients with hypertension. We evaluated demographics, health status, subjective health and mental health, health behaviors, health beliefs, knowledge of hypertension treatment, satisfaction with medical care, and quality of medical care as potential predictors of QoL and perceived QoL in the hypothetical absence of hypertension. RESULTS: Worse financial status, poorer blood pressure control, worse subjective health, mental distress, lack of hypertension diet, and irregular hypertension care were all associated with worse QoL. Worse financial status, poorer blood pressure control, higher body mass index, mental distress, and following a hypertension diet were associated with better QoL in the hypothetical absence of hypertension. CONCLUSIONS: In addition to taking into consideration well known determinants of QoL, such as financial status, health status, and mental health, physicians need to be aware of the potential tradeoff between following medical recommendations that advocate for a strict diet and the impact these have on patients' QoL. Physicians also need to be aware of the effect of imparting information regarding imbalanced blood pressure on perceived QoL. PMID- 17408002 TI - Co-inoculation of an antibiotic-producing bacterium and a lytic enzyme-producing bacterium for the biocontrol of tomato wilt caused by Fusarium oxysporum f. sp. lycopersici. AB - The antifungal compound 2,4-diacetylphloroglucinol-producing bacterium, Pseudomonas fluorescens strain LRB3W1, inhibits the growth of Fusarium oxysporum f. sp. lycopersici, and controls Fusarium wilt of tomato caused by F. oxysporum f. sp. lycopersici. On the other hand, Serratia marcescens strain B2, which produces cell wall-degrading enzyme chitinases, did not inhibit fungal growth and the suppressive effect of strain B2 against tomato Fusarium wilt was less than that of strain LRB3W1. Combined inoculation of strain LRB3W1 with strain B2 was more effective than treatment with strain LRB3W1 alone. When 2,4 diacetylphloroglucinol and the chitinolytic enzymes were applied in combination, a synergistic inhibitory effect against the pathogen was observed. It was possible that bacteria which produce cell wall-degrading enzymes enhanced the biocontrol effect of the antibiotic-producing bacterium against tomato Fusarium wilt. PMID- 17408003 TI - Antibacterial activity of stilbene oligomers against vancomycin-resistant Enterococci (VRE) and methicillin-resistant Staphylococcus aureus (MRSA) and their synergism with antibiotics. AB - Two resveratrol trimers, gnemonol B isolated from Gnetum gnemon and gnetin E obtained from the Gnetum species, were found to exhibit strong antibacterial activities against vancomycin-resistant Enterococci (VRE) and methicillin resistant Staphylococcus aureus (MRSA). The MIC values of gnemonol B against five strains of VRE and nine strains of MRSA were 12.5 and 6.25 microg/ml, respectively. The MIC values of gnetin E against five strains of VRE and nine strains of MRSA ranged from 12.5 to 25microg/ml. These compounds also showed synergistic effects when used in combination with commercially available antibiotics according to the evaluation method using FIC indices. These findings suggested that the application of the test compounds alone or in combination with antibiotics might be useful in controlling and treating VRE and MRSA infections PMID- 17408004 TI - Antibacterial activity of carbon-coated zinc oxide particles. AB - Particles of ZnO coated with carbon (ZnOCC) were prepared and evaluated for their antibacterial activity. ZnO powder and poly(vinyl alcohol) (PVA) (polymerization degree: 2,000-95,000) were mixed at a mass ratio (ZnO/PVA) of 1, and then heated at 500-650 degree C for 3 h under argon gas with a flow rate of 50ml/min. Carbon deposited on the ZnOCC surface was amorphous as revealed by X-ray diffraction studies. The ZnOCC particles maintained their shape in water, even under agitation. The antibacterial activity of ZnOCC powder against Staphylococcus aureus was evaluated quantitatively by measuring the change in the electrical conductivity of the growth medium caused by bacterial metabolism (conductimetric assay). The conductivity curves obtained were analyzed using the growth inhibition kinetic model proposed by Takahashi for calorimetric evaluation, allowing the estimation of the antibacterial efficacy and kinetic parameters of ZnOCC. In a previous study, when ZnO was immobilized on materials, such as activated carbon, the amount of ZnO immobilized was approximately 10-50%, and the antibacterial activity markedly decreased compared to that of the original ZnO. On the other hand, the ZnOCC particles prepared in this study contained approximately 95% ZnO and possessed antibacterial activity similar to that of pure ZnO. The carbon-coating treatment could maintain the antibacterial efficacy of the ZnO and may be useful in the develop-ment of multifunctional antimicrobial materials. PMID- 17408005 TI - Properties of bacterial corrosion of stainless steel and its inhibition by protamine coating. AB - We investigated characteristics of the corrosion of stainless steel specimens by bacteria and the effects of using antimicrobial coating on the surface for inhibiting corrosion. Bacillus sp. 2-A and Staphylococcus sp. 2-1 cells adhered tightly to a stainless steel SUS304 specimen, formed a microcolony or biofilm, and had highly corrosive activities. Microbially influenced corrosion (MC) was observed under or around adhering cells. However, dead cells were markedly less active than viable cells not only in corroding the specimen but also in adhering to its surface. The culture supernatant was not able to induce the corrosion of SUS304 effectively. A protamine coating on the specimen killed bacterial cells only on its surface, interfered with cell adhesion, and inhibited MC. From these results, adhesion of viable cells to the surface of a SUS304 specimen led to the outbreak of MC. Protamine was also found to be an effective substance tested for protecting the specimen from both cell adhesion and surface MC. We suggest that a protamine coating can be applied as a convenient and inexpensive corrosion prevention method. PMID- 17408006 TI - Automatic mapping of viable microbial cells distributed in the surface layer of cotton fabrics. AB - Viable microbial cells distributed in a 130 microim thick surface layer of cotton fabrics were stained with a fluorescent glucose, 2- [N- (7-nitrobenz-2-oxa-1,3 diazol-4-yl) amino] -2-deoxy-D-glucose (2-NBDG), and automatically mapped with an ultra-deep focusing range microscope (UDF) system. The software of the UDF system was upgraded and the number of Candida albicans cells could be counted at a higher precision than before. Bacterial cells of Pseudomonas fluorescens, Serratia marcescens, and Citrobacter freundii, which were smaller than 1-2 microm, were successfully mapped for the first time. These results indicate the practical importance of the present method in the evaluation of the antibacterial properties of fabrics and the efficacy of washing. PMID- 17408007 TI - Construction of Thermokill Database R8100, an expanded version of a microbial thermal death database developed on the basis of information in research papers published from 1981 to 2000. AB - Following the former version of the database on the thermal death of microorganisms, ThermoKill Database R9100, we constructed an expanded version, ThermoKill Database R8100. This database includes the thermal death data together with as many as 70 items of experimental conditions described in research papers which appeared in 23 academic journals published worldwide from 1981 to 2000. The novel version contains 7,454 records as data sets, 12,261 D values, and 2,038 z values. Some statistical data based on the data input in the database are presented. The significance of this novel database in practical use is then discussed. PMID- 17408008 TI - Characteristics of children in residential treatment in New York State. AB - This study addresses three questions about the population of children and families served in the highest level of care in the child welfare system in New York State residential treatment centers (RTCs): (1) How prevalent are emotional and behavioral problems in the youth entering RTCs? (2) Has the proportion of youth with such problems increased compared to 10 years ago? (3) Are there identifiable subgroups of youth entering RTCs? One-fourth of RTC admissions in fiscal year 2001 were randomly selected from a representative sample of 16 RTCs. The study completed a standardized data collection instrument based on a review of agency records, and included information that was known at the time each child was admitted. The results show significant increases compared to t10 years earlier in the proportion of youth with mental health problems and juvenile justice backgrounds. The findings suggest that youth who traditionally have been served by other systems of care are now being served in the child welfare system. The increased treatment needs of these youth and the heterogeneity of the RTC population have important implications for policies, programs, and practice. PMID- 17408009 TI - Birthfamilies as permanency resources for children in long-term foster care. AB - Provisions of the Adoption and Safe Families Act of 1997 mandated shorter time frames for making permanency decisions and facilitating adoption. Yet for many children, foster care continued to be a significant portion of their life experiences. This project explored the potential permanency option of birthfamilies and extended kin for children who languished in foster care while being free for adoption. Eighteen children achieved permanent placement with their birthfamilies. In addition, staff found that although many families could not provide permanent placements, they could offer appropriate relationships with the children. This project team recommends viewing family relationships as an integral component where placement is one option on a continuum that includes letters, phone calls, and visits. PMID- 17408010 TI - Organizational constructs as predictors of effectiveness in child welfare interventions. AB - Organizational context, including line worker characteristics and service settings, may help explain the equivocal findings of intervention studies in the field of child welfare. Yet organizational context has been largely ignored in studies of child welfare interventions. The purpose of this article is to expound upon the likely role of the organizational context in explaining service effectiveness in child welfare. Several bodies of literature within child welfare and human service organization and administration are reviewed and synthesized. A conceptual framework that can be used to guide future child welfare research is then proposed. PMID- 17408011 TI - A clinical consultation model for child welfare supervisors. AB - This article presents findings from a consultation project conducted by faculty from six schools of social work with approximately 150 child welfare supervisors over a two-year period. The purpose of the program was to assist supervisors with their roles as educators, mentors, and coaches for casework staff, specifically in relationship to case practice decisions. The consultation model, the development of the curriculum, the project implementation, and the results of the initial assessment are described. PMID- 17408012 TI - Improving child welfare performance: retrospective and prospective approaches. AB - Some of the key outcome measures used in the first federal Child and Family Service Reviews rely on retrospective cohorts and exclude key portions of the population from the analysis. Most discussions of this issue have focused on the extent to which retrospective measurement is a valid basis on which to judge states' performance (Courtney, Needell, & Wulczyn, 2003). The analyses presented here suggest that in some instances the relative or comparative results of retrospective and prospective measurements exhibit few differences. On the other hand, the analyses also make clear that retrospective measurements have two serious deficiencies in relation to improving performance. First, they are likely to identify the wrong issues, and, second, even when they identify the correct issues they fail to provide information needed to improve performance. This article suggests some practical ways in which the information currently available to child welfare agencies can be used to correct these problems. PMID- 17408013 TI - Domestic violence screening and service acceptance among adult victims in a dependency court setting. AB - Many child welfare systems are unable to effectively identify and address co occurring domestic violence and child maltreatment. In response, the Dependency Court Intervention Program for Family Violence implemented a protocol to identify indicators of domestic violence in families involved with child protection proceedings. This article highlights data that demonstrate the ability of an outreach and screening process to identify adult victims of domestic violence in dependency court and to offer them appropriate intervention services. PMID- 17408014 TI - [Critical evaluation of the oral glucose tolerance test for the diagnosis of insulin resistance in patients with polycystic ovary syndrome]. AB - Oral glucose tolerance test (OGTT) is the most commonly used method to evaluate insulin resistance (IR) in the clinical practice. Our objective was to evaluate the diagnostic utility of fasting tests compared with OGTT tests in women with PCO, and the ability of fasting tests to detect postprandial hyperglycemia. One hundred fourteen women with PCO and 29 normal women were evaluated by a 2 hours OGTT. Fasting plasma insulin (INS) and glucose were measured during the test. GLU:INS ratio (r) (fasting glucose/fasting insulin), HOMA (homeostatic model assessment), QUICKI (quantitative insulin sensitivity check index) (fasting tests), as well as the AUCI (area under the curve of INS) and ISI composite (ISI) (insulin sensitivity index) (OGTT tests), were determined. A significant correlation between fasting tests and OGTT tests was found. Normal fasting tests with abnormal OGTT tests were found in 9 patients. No patient with fasting insulin levels less than 9.9 Ul/ml were IR, and all women with fasting insulin levels over 18.4 UI/ml were classified as having IR. We found glucose levels 120 min post glucose load (G 120) > or = 140 mg/dl in 14 patients (12.2%). Fasting glucose and insulin levels and the fasting tests, were poor predictors of impaired glucose tolerance (IGT) and type 2 diabetes (DBT 2). Thus, fasting tests are useful in the diagnostic of IR in PCO patients. OGTT is necessary when the fasting insulin levels range between 9.9 and 18.4 Ul/ml. Women with PCO should undergo periodic screening for abnormal glucose tolerance. PMID- 17408015 TI - [Survey of risk factors and renal disease in first-degree relatives of dialysis patients]. AB - BACKGROUND: It has been established that first-degree relatives of patients with chronic kidney disease (CKD) have a higher CKD risk than the overall population. This paper deals with the relative frequency of CKD markers and cardiovascular (CV) risk factors within first-degree relatives of ESRD patients in Argentina. METHODS: 810 family members volunteered to participate; of them 668 over 18 ys. old. Trained nurses interviewed them and completed a questionnaire dealing with family history of renal and cardiovascular disease. Blood pressure, urine and blood analysis and anthropometric data were collected. Selected parameters were: smoking habit, presence of high blood pressure, diabetes, hypercholesterolemia, high plasma creatinine and creatinine clearance estimated by MDRD formula, proteinuria and microalbuminuria. In pediatric population, weight and blood pressure parameters were evaluated as percentiles. CKD were classified in stage (National Kidney Foundation). RESULTS: The relative frequencies were: CKD: 29.6%; proteinuria: 13.9%; microalbuminuria: 8.7%. The prevalence values found for main CV risks factors, adjusted by sex and age, were: high blood pressure= 41.8%; overweight/obesity by BMI= 62.1%, hypercholesterolemia= 42.9% and hyperglycemia= 5.2%. Smoking habit was present in 34.8%. IN CONCLUSION: Prevalence of overweight/obesity, hypertension and hypercholesterolemia in first-degree relatives of ESRD patients is higher than previously communicated in studies of national reference populations. Prevalence of CKD is high, estimated as three fold higher than for a general population as reported in poblational studies. These results support the fact that first-degree relatives of ESRD patients, as has been established elsewhere, constitute a population at high risk for developing ESRD. PMID- 17408016 TI - [Mild cognitive impairment. Survey of attitudes of specialists and general physicians. mild]. AB - Mild cognitive impairment (MCI) refers to persons who are slightly cognitively impaired for age but do not meet the criteria for dementia. MCI has been related to a pre-dementia stage of Alzheimer's disease (AD). However, other possible diagnoses such as cerebro-vascular disease, frontotemporal dementia or normal aging have been considered. Diagnosis, etiology and conversion to dementia are a source of ambiguity in MCI. The aim was to evaluate the opinion of experts on dementia and of general practitioners concerning MCI. A total of 24 experts from Argentina and Brazil (16 neurologists and 8 psychiatrists) and 30 general practitioners agreed to reply to a questionnaire on MCI (adapted from Dubois inventory, 2003). Of these, 92% of experts considered MCI as an ambiguous entity, not necessarily as a "pre-dementia" stage; 63% confirmed a tendency to worsen over the time and 83% of experts decided to initiate treatment using cholinesterase inhibitors, memantine and vitamin E. The opinion on MCI was that a priori it is not only an Alzheimer disease pre-dementia stage, but most of them consider the treatment against AD. MCI is a heterogeneous entity that should be classified as an open category and making it necessary to standardize definitions and design diagnosis guides to better understand Alzheimer disease pre-dementia stage. PMID- 17408017 TI - Ketoconazole therapy: an efficacious alternative to achieve eucortisolism in patients with Cushing's syndrome. AB - Cushing's syndrome (CS) is a serious condition requiring drug management in diverse clinical settings. Fifty four patients (44 females, 10 males) with CS, aged 14-63, received ketoconazole (KTZ) prior to surgery (n= 27), as complementary therapy after surgery and/or radiotherapy (n= 16), or as primary treatment (n= 11). It was given at a 600 (500 - 600) mg/day (median - Cl195) maintenance dose for periods ranging from 15 days to 13 years. Clinical signs, hepatic enzymes and urinary free cortisol (UFC) were evaluated before and during KTZ treatment. UFC normalised or decreased to subnormal values in 85% of the patients, in 5 to 150 days after starting treatment; although failing to normalise, UFC decreased to 12-48% of pre-treatment values in the remaining patients. Clinical signs improved throughout. Side effects were adrenal insufficiency (18.5%), reversible hepatic toxicity (11%), allergic skin rash (5.5%) and gastric intolerance (3.7%); in 11% of patients, an "escape phenomenon" was observed. Twenty-four out of the total (44.4%) were treated for prolonged periods, from one up to 13 years. In conclusion, this study confirms that KTZ is an effective and generally well tolerated treatment for CS particularly: a) shortly before surgery, b) because of persistent hypercortisolism after surgery or awaiting the results of radiotherapy, c) as a reasonable option in patients with CS of unknown aetiology and, d) as long-term therapy in any case of unsolved hypercortisolism after failure of current treatments. PMID- 17408018 TI - Genotypes of vitamin D and estrogen receptors in pre and perimenopausal women from Cordoba, Argentina. AB - The aim of this study was to determine the frequency of vitamin D receptor and estrogen receptor genotypes and their relationship with the lumbar spine or femoral neck bone mineral density in healthy pre and perimenopausal women from Cordoba (Argentina) and adjacent areas. Genotypes were assessed by restriction fragment length polymorphism-polymerase chain reaction technique. Bsm I and Fok I for vitamin D receptor gene and Xbal and Pvull for estrogen receptor gene were used as restrictases. Two hundred and ten healthy pre and perimenopausal women were recruited and analyzed by age. Calcemia and serum parathyroid hormone did not change, but serum P and beta-CrossLaps decreased with age. Femoral neck bone mineral density decreased significantly after 30 years old. Vitamin D receptor and estrogen receptor genotype frequencies were similar to those from other Caucasian women. No association between vitamin D receptor and estrogen receptor genotypes with the lumbar spine or femoral neck bone mineral density has been detected. Analysis of interaction between vitamin D receptor and estrogen receptor genes using covariates such as age, height and body mass index did not show any influence of the combination of those genotypes on bone mineral density. Lifestyle, smoking and alcohol intake had no effect on lumbar spine and femoral neck bone mineral density. To conclude, these data do not support the hypothesis that vitamin D receptor and estrogen receptor genotypes influence on lumbar spine and femoral neck bone mineral density in healthy pre and perimenopausal women from this area of Argentina. PMID- 17408019 TI - [Embolism predictors of infective endocarditis]. AB - The embolic event (EE) increases the morbidity and mortality of infective endocarditis (IE). Prevalence of EE ranges between 22% and 50%, death rates being up to 25% of patients. EE may occur prior to diagnosis, during treatment or afterwards. The objective of this study was to evaluate the demographic, clinical, microbiological, echocardiographic and therapeutic characteristics in patients suffering from IE (with or without emboli) in order to determine predictors for EE. A descriptive study based on observations of patient population diagnosed with IE was conducted at the Hospital Italiano of La Plata during the period March 1996 - December 2004. Fifty-three patients with IE were analyzed (35 without EE and 18 with EE) in retrospect. We found that the presence of vegetations in the transthoracic (TTE) and/or transesophagic (TEE) echocardiographies at the time of diagnosis, the size > or = 10 mm and the compromise of the native mitral valve were the variables that showed significant statistical association with EE to be considered as predictors. The size _ 10 mm was the only variable associated with EE in the logistic regression analysis. During the elective antibiotic treatment, there was a reduction in EE, without their being present from the second week onwards. PMID- 17408020 TI - [Reversion of the immunological eclipse and therapeutic vaccination against cancer in an experimental model]. AB - Although animals can be prophylactically immunized against the growth of tumor implants, most of the attempts to use immunotherapy to cause the regression of animal and human tumors once they become established have been unsuccessful. To understand the nature of this refractoriness we have studied a methylcholanthrene induced and strongly immunogenic murine fibrosarcoma. In our model, the onset of this refractoriness was associated with the beginning of an immunosuppressive state known as "immunological eclipse" characterized by a loss of the antitumor immune response when tumor grows beyond a critical size. This immunological eclipse was accompanied by the emergence of a systemic inflammatory condition. Treatment of tumor-bearing mice with a single dose of a synthetic corticosteroid, dexamethasone (DX), reduced significantly all parameters of systemic inflammation and simultaneously reversed the immunological eclipse. The reversion of the eclipse upon DX treatment was not curative itself, but allowed an immunological therapy based in dendritic cells pulsed with tumor antigens, which was itself absolutely ineffective, to exert a significant inhibitory effect against an established growing tumor. The two-step schedule using an anti-inflammatory treatment to reverse the immunological eclipse plus a dendritic cell-based vaccination strategy aimed to stimulate the antitumor immune response, could serve eventually as a model of immunotherapy against animal and human tumors. PMID- 17408021 TI - [Reactive hemophagocytic syndrome in critical care patients. Report of 4 cases]. AB - Reactive hemophagocytic syndrome or hemophagocytic lymphohistiocytosis comprises a variety of disorders, many of them associated with infection. It is characterized by hemophagocytosis, with cytopenia involving at least two cellular lines, increase in cytokines and serum ferritin. The clinical course resembles sepsis, sharing similar physiopathological features. We propose that hemophagocytic syndrome is an underdiagnosed entity in the critical care setting; simple tests aid to identify which patients should undergo diagnostic procedures. We discuss current therapeutic approaches. PMID- 17408022 TI - [Severe osteoporosis with vertebral crushes in juvenile dermatomyositis. Effect of oral alendronate therapy]. AB - Glucocorticoids are used for the treatment of inflammatory and autoimmune diseases, cancer, and in prevention of organ rejects. A frequent secondary effect of longterm treatment with corticoids is the loss of bone mass, caused by several mechanisms: decrease in the intestinal calcium absorption, increase of the renal calcium excretion at the distal renal tubule, suppressive effect on the osteoblast and also in apoptosis of osteoclasts, inhibition in local production of IGF I (Insulin-like growth factor) and IGFBPs (binding IGF I proteins necessary for bone metabolism), and decrease on osteocalcin production. Longterm treatment with corticoids is associated with osteoporosis and vertebral fractures. To improve this condition, treatment with bisphosphonates has been proposed. We present here a clinical case of a girl with dermatomyositis and severe osteoporosis with vertebral crushes, who responded well to oral bisphophonate treatment. PMID- 17408023 TI - [A prevalent genetic variety of UDP-glycuronosyl transferase predicts high risk of irinotecan toxicity]. AB - The advances in genetics and molecular biology have raised new areas in medicine, such as pharmacogenomics, which tries to predict drug responses and toxicities based on the individual genetic variability, describing the so called: pharmacogenomic syndromes. Oncology would find this development extremely useful because of the severe toxicity of chemotherapy. There are a lot of genetic loci under investigation for their potential in predicting drug toxicity, but only three of them have showed clinical usefulness up to now. In particular, quantification of the number of thymine-adenine (TA) dinucleotics in the promoter region of the UDP-glucuronosyl-transferase 1A1 enzime (TA indel) proved to be capable of predicting severe neutropenia in patients exposed to intermediate or high doses of irinotecan. Herein we report a case of a patient with small cell lung cancer who suffered severe hematological and gastrointestinal toxicity after being treated with relatively low doses (65 mg/m(2)) of irinotecan and whose leucocyte DNA analysis showed the presence of seven TA repetitions in both alleles. This case is an example of the clinical applicability and the utility of the test as a toxicity predictor. We also discuss the clinical decisions that may be taken with these patients. PMID- 17408024 TI - [Tuberculous Spondylodiscitis]. PMID- 17408025 TI - [Acquired hepatocerebral degeneration]. PMID- 17408026 TI - [The role of dendritic cells in the infection by HIV and HCV]. AB - Dendritic cells are most important as antigen presenting cells during the induction of an effective immune response. Therefore, it is important to study their role during the generation of persistent or chronic viral infections, such as HIV or HCV infection. In this review we shall describe the phenotypic and functional characteristics of the different classes of dendritic cells and of their membrane receptors. Their participation in defence or facilitation mechanisms involved in the immune response against these viruses will be discussed. It is important to take this knowledge into account when trying to design therapeutic strategies for protection or reconstruction of the immune system that may be altered as a consequence of infection with HIV or HCV. PMID- 17408027 TI - [High altitude pulmonary edema. An experiment of nature to study the underlying mechanisms of hypoxic pulmonary hypertension and pulmonary edema in humans]. AB - High altitude constitutes an exciting natural laboratory for medical research. Over the past decade, it has become clear that the results of high-altitude research may have important implications not only for the understanding of diseases in the millions of people living permanently at high altitude, but also for the treatment of hypoxemia-related disease states in patients living at low altitude. High-altitude pulmonary edema (HAPE) is a life-threatening condition occurring in predisposed, but otherwise healthy subjects, and, therefore, allows to study underlying mechanisms of pulmonary edema in humans, in the absence of confounding factors. Over the past decade, evidence has accumulated that HAPE results from the conjunction of two major defects, augmented alveolar fluid flooding resulting from exaggerated hypoxic pulmonary hypertension, and impaired alveolar fluid clearance related to defective respiratory transepithelial sodium transport. Here, after a brief presentation of the clinical features of HAPE, we review this novel concept. We provide experimental evidence for the novel concept that impaired pulmonary endothelial and epithelial nitric oxide synthesis and/or bioavailability may represent the central underlying defect predisposing to exaggerated hypoxic pulmonary vasoconstriction and alveolar fluid flooding. We demonstrate that exaggerated pulmonary hypertension, while possibly a condition sine qua non, may not be sufficient to cause HAPE, and how defective alveolar fluid clearance may represent a second important pathogenic mechanism. Finally, we outline how this insight gained from studies in HAPE may be translated into the management of hypoxemia related disease states in general. PMID- 17408028 TI - [HBV and HCV co-infections in HIV positive patients in the "HAART era": new challenges]. AB - Co-infections with HIV and HCV/HBV are frequently found due to the similar routes of transmission (sexual, parenteral and vertical). Since the introduction of highly active antiretroviral therapy (HAART) there has been a notably decrease in patients morbidity and mortality, nevertheless with the prolonged survival, many of these patients are at risk of developing chronic complication, secondary to the infection of hepatotropic viruses. End stage liver disease is one of the main causes of morbid-mortality among HIV patients in developed countries. Nowadays there are new available therapies, diagnostic and follow up techniques for HBV and HCV, what provides a better control of both co-infections. PMID- 17408029 TI - [2006 Nobel prize in physiology or medicine. The silence of genes]. PMID- 17408030 TI - [Difficulties for the detection of multiple genotypes of hepatitis C virus]. PMID- 17408031 TI - [Treatment in the time of cholera]. PMID- 17408032 TI - [Smelling under mechanical assistance]. PMID- 17408033 TI - [Occupational exposure to hepatitis C virus]. PMID- 17408034 TI - [The "mushroom" on the cheek]. PMID- 17408036 TI - [Functional gastrointestinal disorders: nutrition tips instead of medication]. PMID- 17408035 TI - [IQWIG Preliminary Report: Diuretics are the best antihypertensives. How does it go on with the high blood pressure therapy? (interview)]. PMID- 17408037 TI - [How psychological stress affects digestion. Disordered communication between the abdomen- and head brain]. PMID- 17408038 TI - [Medical care of circumcised women. How genital mutilation injures body and soul (interview)]. PMID- 17408039 TI - [Lung treated instead of buttocks]. PMID- 17408040 TI - [By what means does the rheumatic patient with gastrointestinal risks fare best?]. PMID- 17408041 TI - [Is the therapy compliance or personality of the patient decisive?]. PMID- 17408042 TI - [Wound care and minor surgery. What the family physician can do]. PMID- 17408043 TI - [Soft tissue infections treatable in the doctor's office]. AB - A good number of infections requiring only minor surgery can be treated in the GP's office, simply and adequately. Of importance, however, is the physician's ability to recognize when a patient needs to be referred to a specialist. Provided the principles listed here are adhered to, treatment is usually successful. All other courses indicate a multifactorial pathology and belong in the hands of an experienced specialist. PMID- 17408044 TI - [Diagnosis and treatment of warts, corns, and clavi]. AB - The term keratosis covers a variety of growths such as warts and pressure-induced callosities. Warts develop in the absence of pressure, and often clear up again after a certain period, usually without treatment. Clavi can develop as a result of wearing poorly fitting shoes, but also deformed toes or other anatomical deformities affecting the foot. The resulting non-anatomical stressing leads to the formation of pressure callosities and even to pressure sores. PMID- 17408045 TI - [Diagnosis and treatment of paronychia]. AB - Paronychia is a purulent infection of the lateral fingertip, and represents the most common infectious disease affecting the hand.The typical germ is Staphylococcus aureus, while a mixture of bacteria or pathogens of a different kind are usually seen in patients with immunodeficiency. Treatment ranges from bathing the finger in an antiseptic solution to lancing the abscess. PMID- 17408046 TI - [Management of acute wounds]. AB - Uncomplicated wounds can be treated immediately in the general practitioner's office. Management by a specialist is necessary only in the case of wounds involving the eyes, nose or mouth, or in the presence of other unusual features. PMID- 17408047 TI - [My child is so restless]. AB - Hyperkinetic or attention deficit hyperactivity disorders (ADHD) are characterized by three symptoms: attention deficit, hyperactivity and impulsiveness. For some patients, intensive, continuous counselling or behaviour therapy leads to adequate success. If this is not effective, drug treatment using stimulants such as methylphenidate or the selective norepinephrine reuptake inhibitor atomoxetine is indicated. PMID- 17408048 TI - [Emergency checklist: CS distortion]. PMID- 17408049 TI - [Inflammation of the glans penis]. PMID- 17408050 TI - [From the choice to the accounting -- tips about IGeLn]. PMID- 17408051 TI - The influence of micronutrients on cognitive function and performance. AB - There has been much media speculation (often sensationalist and conflicting) regarding the potential influence of micronutrients on cognitive function and performance. Our aim was to identify the micronutrients specifically implicated in cognitive function and to review the literature to identify original sources underlying the media coverage. Literature searches were carried out to identify recent clinical trials, reviews, editorials and meetings describing the biochemical and physiological role of individual micronutrients. No attempt was made to grade the evidence. The searches confirmed that the water-soluble vitamins (B group and C), together with the minerals, calcium, magnesium and zinc, are most relevant to cognitive performance. Clinical evidence revealed that marginal deficiencies of one or more of these micronutrients are not uncommon, even in the developed countries, and that such deficiencies may affect cognitive performance, especially in vulnerable groups such as the elderly and those individuals who are exposed to occupational pressures and a stressful lifestyle. PMID- 17408052 TI - Comparison of the nutritional values of follow-on formulae available in Italy. AB - If breast milk is not available during complementary feeding, 'follow-on' formula milk ensures a balanced intake of nutrients and is undoubtedly more suitable for infants than cows' milk. Formula milk provides a lower intake of proteins and saturated fats and a higher concentration of polyunsaturated fats, iron and zinc than cows' milk. In particular, delaying the introduction of cows' milk until the end of the first year of life, or even until after the second year, appears to play a fundamental role in the prevention of iron deficiency anaemia. It may also prevent excess weight gain and obesity and the development of immunoallergic symptoms, and may be important for neurobehavioural development. More than 30 varieties of follow-on formula milk are currently available in Italy alone. This review outlines the nutritional differences between breast, formula and cows' milks, and compares different brands of formula milk. PMID- 17408053 TI - Obstructive sleep apnoea, cigarette smoking and serum testosterone levels in a male sleep clinic cohort. AB - This study evaluated the impact of obstructive sleep apnoea (OSA) and smoking on total serum testosterone levels in 96 men (mean age 43.3 years; range 25 - 60 years) attending a sleep clinic. Fifty-five men (57.3%) had OSA, defined as an apnoea-hypopnoea index of > or = 15 events/h, recorded during overnight polysomnography, and 42 (43.8%) were current smokers. Mean serum total testosterone levels were significantly lower in OSA subjects (3.4 ng/ml) than in non-OSA subjects (3.9 ng/ml), whereas no significant difference was observed between current smokers and nonsmokers. In a multiple linear regression analysis, serum testosterone was negatively correlated with body mass index and the apnoea hypopnoea index, but not with age and pack-years of smoking. Our results support previous observations regarding testosterone levels in men with OSA, but, contrary to some earlier reports, there was no positive relationship between smoking and total testosterone in the present cohort. PMID- 17408054 TI - Alcohol and acute myocardial infarction. AB - A number of studies from different countries and several large-scale meta analyses have reported reduced coronary heart disease rates among those regularly consuming mild to moderate amounts of alcohol compared with those abstaining from alcohol. In contrast, various studies have also reported that heavy alcohol consumption promotes the progression of atherosclerosis and that binge drinking might trigger embolic stroke and acute myocardial infarction. We discuss the association between alcohol consumption and acute myocardial infarction on the basis of evidence from literature published recently. Alcohol consumption has both favourable and unfavourable effects on metabolism, lipid profile, blood coagulation and fibrinolysis, blood pressure and vascular tone depending on the amount of alcohol consumed and the way that it is drunk (i.e. drinking habits). We conclude that it is extremely important to warn people of the risks associated with binge drinking and to encourage them to remain within the recommended safe limits for alcohol consumption. PMID- 17408055 TI - Effect of nicardipine on haemodynamic and bispectral index changes following endotracheal intubation. AB - We investigated the effect of IV nicardipine on haemodynamic and bispectral index responses to the induction of general anaesthesia and intubation. Forty patients were randomly allocated to two groups of 20 to receive normal saline or nicardipine 15 microg/kg IV 30 s after induction. Ninety seconds later, tracheal intubation was performed. Systolic blood pressure, heart rate and bispectral index were measured at baseline, 1 min after induction, pre-intubation, and every minute until 5 min after endotracheal intubation. Rate-pressure product values were calculated. In the nicardipine group, systolic blood pressure decreased compared with the control group, and heart rate increased comparedwith the control group. Bispectral index and rate-pressure product showed no differences between the two groups. In conclusion, the administration of 15 microg/kg nicardipine IV does not affect anaesthetic depth in response to the induction of general anaesthesia and intubation. PMID- 17408056 TI - The effects of calcium channel blockers are not related to their chemical structure in the collar model of the rabbit. AB - Placing a silicone collar around the rabbit carotid artery induces intimal thickening, an early stage in atherosclerosis and restenosis. We investigatedwhethertreatment with oral pranidipine, a new potent, long-lasting dihydropyridine calcium channel blocker (CCB), inhibited collar-induced intimal thickening in addition to the changes in vascular reactivity usually observed in this model. Pranidipine treatment did not inhibit collar-induced intimal thickening. Placing the collar around the carotid artery resulted in the characteristic changes in vascular reactivity, such as increased sensitivity to 5 hydroxytryptamine. Treatment with Nomega-nitro-L-arginine (100 microM) and pranidipine, however, did not affect collar-induced changes in vascular reactivity. From results of this and previous studies, we conclude that pranidipine does not prevent collar-induced intimal thickening or collar-induced changes in vascular reactivity. Not all CCBs prevent collar-induced intimal thickening, suggesting that the effects of these agents are not related to their chemical structure and/or their calcium channel-blocking actions. PMID- 17408057 TI - Barrier function of the gut and multiple organ dysfunction after cardiac surgery. AB - We studied the influence of sequential involvement of the gastrointestinal (GI) tract on the development of multiple organ dysfunction (MOD) after cardiopulmonary bypass (CPB). One hundred and forty-six patients undergoing elective cardiac surgery were included in this prospective observational study. Standardized oral inert-sugar tests (sucrose, lactulose, mannitol, sucralose) were performed before and after CPB in different patients. Enzyme-linked immunosorbent assay of plasma levels of endotoxin core antibodies (EndoCAb) were performed peri-operatively. The functional mucosal surface was calculated from the amount of mannitol absorbed from the GI tract. Lower urine concentrations of absorbed mannitol were observed pre-operatively in patients developing MOD. In binary logistic regression this was an independent parameter. Decreased plasma concentrations of EndoCAb after surgery were seen in every patient, but were more significant in patients developing MOD. A reduced pre-operative functional mucosal surface may predict the early occurrence of MOD after surgery. PMID- 17408058 TI - Effects of Astragalus polysaccharides and astragalosides on the phagocytosis of Mycobacterium tuberculosis by macrophages. AB - The herb Astragalus membranaceus is used in traditional Chinese medicine to boost immunity. This study investigated the effects of Astragalus polysaccharides (APS) and astragalosides (AS) on the phagocytosis of Mycobacterium tuberculosis by macrophages. Peritoneal macrophages were obtained by peritoneal lavage from mice stimulated by starch gravy culture medium and cultured with M. tuberculosis and varying concentrations of APS and AS. Phagocytotic activity was measured using a real-time polymerase chain reaction assay to detect M. tuberculosis DNA. Levels of interleukin-1beta, interleukin-6 and tumour necrosis factor-a secreted by activated macrophages in the culture supernatant were determined using an enzyme linked immunosorbent assay. Macrophage phagocytotic activity and secreted cytokine levels were significantly increased after treatment with APS and AS. This study provides evidence that APS and AS have strong promoting effects on the phagocytosis of M. tuberculosis by macrophages and the secretion of interleukin lbeta, interleukin-6 and tumour necrosis factor-alpha by activated macrophages. PMID- 17408059 TI - Evaluation of driver stress in a motor-vehicle driving simulator using a biochemical marker. AB - The purpose of this study was to evaluate salivary amylase activity (sAMY) as an indicator of the acute psychological effects of driving. The influence of the operation of a device not directly associated with driving was also estimated using a car navigation system. The psychological effects of driving were examined using sAMY analysis, oculomotor angle and subjective evaluation with a questionnaire, and the methods were compared. The change in sAMY over time was analysed before and during driving. The results indicate that the psychological effect of driving-induced stress, a condition that cannot be easily detected or recognized by a subjective evaluation, is quickly quantified using sAMY. Moreover, the results suggest that the operation of a non-driving-related device may also reduce the driver's capacity to concentrate on driving. PMID- 17408060 TI - Use of tissue Doppler imaging following coronary artery bypass surgery. AB - This study assessed the effect of coronary artery bypass grafting (CABG) on myocardial systolic functions using tissue Doppler imaging (TDI). Fourteen patients (three women and 11 men) who had undergone isolated coronary bypass surgery were included in the study. Their mean age was 61 +/- 8 years. TDI systolic velocity measures were obtained from four different sites on the left ventricular wall (anterior, septal, lateral and inferior) at the papillary muscle level in the parasternal short axis view before CABG, and then at 1 and 6 weeks post-operatively. There were significant increases in the myocardial wall velocities at all left ventricular sites 1 week after CABG. This increase persisted to week 6 after CABG, but the velocities were lower than week 1 values. We conclude that the ischaemic myocardium responded to surgical revascularization with marked increases in myocardial segmental systolic velocities in the early post-operative period. PMID- 17408061 TI - Nifedipine, a calcium-channel blocker, inhibits advanced glycation end-product induced expression of monocyte chemoattractant protein-1 in human cultured mesangial cells. AB - The interaction between advanced glycation end-products (AGEs) and their receptors mediates the progressive alteration in renal architecture and loss of renal function in diabetic nephropathy. This study investigated whether nifedipine, a widely used anti-hypertensive drug, suppresses expression of monocyte chemoattractant protein-1 (MCP-1), a chemokine that mediates the recruitment of monocytes to inflammatory sites, in AGE-exposed human cultured mesangial cells. Cells were treated with 100 microg/ml AGE-bovine serum albumin (BSA) or non-glycated BSA in the presence or absence of 1 microM nifedipine or 50 nM diphenylene iodonium, an inhibitor of reduced nicotinamide-adenine dinucleotide phosphate oxidase, for 4 or 24 h. Expression of MCP-1 mRNA was measured using a semi-quantitative reverse transcription-polymerase chain reaction; MCP-1 protein production was measured using an enzyme-linked immunosorbent assay. AGEs significantly increased both MCP-1 mRNA expression and protein production in mesangial cells; this increase was blocked by both nifedipine and diphenylene iodonium. These results suggest that nifedipine could play a protective role against early diabetic nephropathy by suppressing MCP-1 overexpression via blockade of AGE signalling in mesangial cells. PMID- 17408062 TI - Arterial stenosis detected by digital subtraction angiography and its relationship with the Oxfordshire Community Stroke Project classification. AB - The aim of this study was to assess extra-and intracranial arterial stenosis in first-ever stroke patients and to analyse the relationship between arterial stenosis and the Oxfordshire Community Stroke Project (OCSP) classification. Chinese patients (n = 817) were classified according to OCSP criteria, and arterial abnormalities were evaluated in 342 of the patients by digital subtraction angiography. Prevalence of significant stenosis in the extracranial internal carotid artery was highest among total anterior circulation infarct patients and lowest among patients with lacunar infarct. There was no evidence that the frequency of patients with intracranial arterial significant stenosis was different among OCSP subtypes. The results suggest that extracranial internal carotid artery stenosis was significantly related to OCSP subtype and that patients with total anterior circulation infarct were more likely to have extracranial internal carotid artery stenosis than patients having partial anterior circulation infarct, posterior circulation infarct or lacunar infarct. PMID- 17408063 TI - Hypocapnic alkalosis enhances oxidant-induced apoptosis of human alveolar epithelial type II cells. AB - Apoptosis of alveolar epithelial type II (AEC-II) cells induced by reactive oxygen species (ROS) contributes to extensive alveolar damage during acute lung injury. Hypercapnic acidosis and hypocapnic alkalosis are known to modulate ROS mediated lung damage. This study assessed the effects of acid-base balance disturbances on hydrogen peroxide (H2O2)-induced apoptosis of the AEC-II-like human cell line A549, which was cultured under different conditions of pH and CO2 tension (normal pH and CO2, hypercapnic acidosis, metabolic acidosis, hypocapnic alkalosis and metabolic alkalosis). H2O2-induced apoptosis was assessed by a dye uptake bioassay and induction of caspase activity, which were quantified using analytical digital photomicroscopy. Acidosis or alkalosis of the culture medium alone did not induce A549 cell apoptosis. Hypocapnic alkalosis significantly increased H2O2-induced apoptosis and caspase activation of A549 cells. Metabolic alkalosis non-significantly increased H2O2-induced A549 cell apoptosis and caspase activation. These data suggest that hypocapnic alkalosis intensifies oxidative-induced apoptosis of alveolar epithelial cells. PMID- 17408064 TI - Apolipoprotein E and first-ever ischaemic stroke in Greek hospitalized patients. AB - The association between apolipoprotein E (ApoE) polymorphism and stroke is still controversial. This study investigated the potential association between ApoE genotypes and stroke subtypes, and risk factors for ischaemic stroke in Greek patients hospitalized with their first-ever ischaemic stroke. One hundred patients (70 men and 30 women; mean age +/- SD 60.7 +/-9.8 years) were included in the study. The control group comprised 96 age- and sex-matched healthy volunteers. Cerebral infarction was classified as atherothrombotic, cardioembolic or lacunar small-vessel stroke. The three common ApoE alleles (E2, E3 and E4) were determined using the semi-nested polymerase chain reaction. No significant difference in the ApoE alleles was found between patients and controls. Similarly, there was no significant association between ApoE alleles and stroke subtypes, common risk factors for ischaemic stroke and neck vessel stenosis. Although the sample size was small, these results do not support a role for ApoE polymorphism in the pathogenesis of ischaemic stroke. PMID- 17408065 TI - Atrial fibrillation after off-pump coronary artery surgery: a prospective, matched study. AB - The effect of cardiopulmonary bypass and myocardial ischaemia on the occurrence of atrial fibrillation (AF) after coronary artery bypass graft (CABG) was studied in 136 patients undergoing off-pump CABG who were matched for age and number of distal anastomoses with 136 patients undergoing on-pump CABG. Possible risk factors for post-operative new-onset AF were recorded. AF occurred in 64 (24%) of the 267 patients for whom data could be analysed. AF occurred in 29 patients (22%) in the off-pump group versus 35 (26%) in the on-pump group, but this difference was not statistically significant. On univariate analysis, age and length of hospital stay were significant risk factors for the occurrence of AF. In a multivariate analysis that included operative technique, age was found to be the only significant risk factor. In conclusion, the occurrence of AF after CABG does not depend on the type of operation. PMID- 17408066 TI - Ocular bleeding related to warfarin anticoagulation in patients with mechanical heart valve and atrial fibrillation. AB - We investigated the incidence of ocular bleeding in patients taking anticoagulant therapy (warfarin) and its association with other related factors. We examined 210 patients taking warfarin and 210 gender- and age-matched controls for ocular bleeding. Patients and controls were examined by external ocular examination and fundoscopic examination. The incidence of ocular bleeding was 11.4% in patients and 3.8% in controls. It was five times higher in patients with hypertension than in other patients. The incidence of ocular bleeding was higher in older than in younger patients. No association was found between ocular bleeding and factors such as gender, international normalized ratio, duration of warfarin therapy, concomitant aspirin use and diabetes mellitus. Thus, warfarin therapy increases the frequency of ocular bleeding. The higher incidence of ocular bleeding in the patients with hypertension and advanced age should be kept in mind and this subgroup of patients taking warfarin should be monitored closely. PMID- 17408067 TI - Use of a geographic information system in an epidemiological study of measles in Istanbul. AB - A geographic information system was used in an epidemiological study of measles in Istanbul, Turkey. The aim was to obtain more reliable results and to increase the facilities for spatial monitoring in epidemiological studies. The Gaziosmanpasa district of Istanbul was selected as it experiences the highest level of migration, and currently has the highest population growth rate of the city's districts. The epidemic distribution of measles was investigated on the basis of examination of 320 children. The number, geographical distribution, complications and time of measles outbreaks were analysed and represented in thematic maps (a small selection is shown in this paper). The results were evaluated and, in view of these results, new vaccination plans were arranged. PMID- 17408068 TI - The frequency of symptoms in transient ischaemic attack: analysis of Nanjing Stroke Register Program data. AB - The clinical characteristics of 172 patients experiencing single or multiple (two or more) transient ischaemic attacks (TIAs) were investigated. Risk factors, clinical manifestations and vascular examinations were compared. Intracranial and extra-cranial large-artery disease (stenosis > or = 50%) occurred more frequently in multiple-TIA patients than in single-TIA patients, and weakness and atrial fibrillation were observed more frequently in single TIA than in multiple TIA patients. The type of aetiological examination performed in TIA patients should depend on the frequency of attacks. PMID- 17408069 TI - Progressive supranuclear palsy-like parkinsonism resulting from occupational exposure to lead sulphate batteries. AB - A 53-year-old man who had worked for 17 years manufacturing car batteries, with overt exposure to lead, developed a clinical picture initially characterized by signs of parkinsonism, followed by atypical signs such as loss of memory, reduction of eye movement, dysarthria, chorea-like dyskinesia and sexual impotence. The diagnosis of atypical parkinsonism was eventually changed to progressive supranuclear palsy-like parkinsonism. The patient was treated with various anti-Parkinson's disease drugs, including levodopa, with modest improvement. The symptoms deteriorated progressively, leading to permanent occupational disability with noticeable limitation of daily activities. Toxicological studies revealed abnormally high blood levels of lead. Discontinuation of lead exposure was followed first by clinical stabilization and then steady improvement. This case confirms recent reports that link exposure to lead and its compounds with degenerative diseases of the central nervous system, such as Parkinson's disease. PMID- 17408070 TI - Estrogen receptors--known and unknown biological functions. AB - The present literature review includes the data available from 1995 to 2005 about the biological functions of estrogen receptors (ERs) and their role in tumorigenesis. The domain organisation,and cell and tissue localization of two estrogen receptors are described. The article reviews the well known function of these receptors as transcription factors and their less familiar extracellular "non-genome" effects. Because of their stimulating effect on cell proliferation estrogens are supposed to actively participate in the genesis and development of some malignant conditions. We draw particular attention to the significance these estrogen receptors have for the onset and development of tumors outside the reproductive system. Knowledge of the receptor mechanisms of estrogenic action may enhance the diagnostic and therapeutic opportunities to treat a wide range of neoplastic lesions. PMID- 17408071 TI - Current knowledge of Aronia melanocarpa as a medicinal plant. AB - Aronia melanocarpa, native to eastern North America, has become popular in Eastern Europe and Russia. Aronia melanocarpa fruits are one of the richest plant sources of phenolic substances, mainly anthocyanins--glycosides of cyanidin. Anthocyanins are water soluble pigments accounting for the dark blue and even black color of the fruits. Administered orally they can be absorbed as intact glycosides. Aronia melanocarpa fruit juice and anthocyanins derived from the fruits have been studied intensively for the last 15 years. Most of the effects of Aronia melanocarpa anthocyanins are due to their high antioxidative activity. Our investigations have demonstrated a remarkable hepatoprotective, a very good gastroprotective and a pronounced anti-inflammatory effect of Aronia melanocarpa fruit juice in rats as well as a bacteriostatic activity in vitro against Staphylococcus aureus and Escherichia coli and an antiviral activity against type A influenza virus. Research of other authors has demonstrated that Aronia melanocarpa anthocyanins can normalize the carbohydrate metabolism in diabetic patients and in streptozotocin-diabetic rats, have an in vitro antimutagenic activity and exhibit a distinct immunomodulatory activity in human lymphocyte cultures and in patients with breast cancer, suppress the growth of human HT-29 colon cancer cells, inhibit the N-nitrosamine formation in rats and decrease the toxicity and cumulation of cadmium in liver and kidneys. Currently, there are no data in literature about any unwanted and toxic effects of Aronia melanocarpa fruits, juice and extracts. PMID- 17408072 TI - Correlation of frequency of exacerbations with the BODE index in COPD patients. AB - BACKGROUND: In recent research B. Celli et al. (N Engl J Med 2004;350:1005-12) proposed a complex grading system (which they designated by BODE) for predicting the mortality risk in chronic obstructive pulmonary disease (COPD). OBJECTIVE: The aim of the study was to compare the BODE index in COPD patients with frequent exacerbations and in those with infrequent exacerbations. PATIENTS AND METHODS: A 1-year retrospective study was performed in 76 patients with COPD of mean (Sx) age of 59.1 (8.6) years, forced expiratory volume in 1 second % predicted (FEV1%) = 35 (13)%. Exacerbations were ascertained retrospectively by patient interview and from records of treatment such as healthcare databases; the effect of frequent or infrequent exacerbations (> or < 2.3 per year) on the BODE index was examined. RESULTS: A total of 178 exacerbations were recorded. Exacerbation frequency was correlated significantly to BODE index (r = 0.36; P = 0.002). There was a statistically significant difference in the BODE index score (5.6 +/- 2.5 vs 4.1 +/- 1.5; P = 0.002) between the frequent and infrequent exacerbators. CONCLUSION: COPD patients who experienced frequent exacerbations in a previous year have significantly higher BODE score than those who experience infrequent exacerbations. PMID- 17408073 TI - Assessment of the effect of palliative radiotherapy for cancer patients with intracranial metastases using EORTC-QOL-C30 questionnaire. AB - Assessment of quality of life (QOL) is of crucial importance when assigning palliative radiotherapy of brain metastases in cancer patients. AIM: To investigate the influence of whole brain radiotherapy on brain symptoms in patients with cerebral metastases; to evaluate their quality of life before and after radiation therapy using the EORTC-QOL-C30 questionnaire, as well as its influence on patients' survival. PATIENTS AND METHODS: Sixty-five patients with various locations of the primary tumor and brain metastases were included in this study. All of them underwent radiotherapy with cobalt unit. The realized dose of the whole brain was above 30 Gy: 10 x 3 Gy or 15 x 2 Gy. The patients filled in the EORTC-QOL-C30 questionnaire before radiation, at the end of the radiotherapeutic course and a month after it. Clinical characteristics of patients before and after radiotherapy were compared and assessed. All patients were treated with radiotherapy and concurrent corticosteroid treatment. RESULTS: The mean age of the patients was 53 +/- 7.8 years and the median survival was 6.6 months for lung cancer patients and 9.8 months for breast cancer patients. Each EORTC-QOL-C30 questionnaire assessed the patients in three main aspects: functional aspects, general symptoms and global health. Improvement was reported (p < 0.001) for functional indicators and health related quality of life (HRQOL). Several symptoms did not change significantly--financial difficulties, dyspnea and diarrhea. CONCLUSIONS: Whole brain radiation of cancer patients with cerebral metastases is very well tolerated. Side effects are frequently met and can be compensated by applying steroids. The assessment of quality of life gives information on patients' improvement which is more substantial in functioning, symptoms and global health. This study is a precondition for future investigations of the effect of whole brain radiation on cancer patients' quality of life. PMID- 17408074 TI - Transient ischaemic attacks and minor strokes (analyses and clinically significant comparisons). AB - The transient ischaemic attacks (TIA) and minor strokes are independent predictors of disabling strokes with a high medical and social value. PURPOSE: Analysis and comparison of the data from the clinical monitoring of TIA and minor stroke patients in correlation with the different duration of the transient neurological deficit. PATIENTS AND METHODS: 234 patients were monitored clinically in the Clinic of Cerebrovascular Diseases, University Hospital "St. George"-Plovdiv between 2002 and 2004. Clinical data were collected for 79 patients who met the clinical criteria for TIA and 155 patients who met the clinical criteria for minor stroke. Our protocol included medical history, cardiac and neurological examinations, assessment of cerebrovascular risk factors and laboratory tests. The instrumental assessment included CT scan, MRI and Doppler examination. The data were processed using descriptive statistics, non parametric methods and charts. RESULTS: The comparative analysis between the TIA and minor stroke patients shows a significant difference only in the number of registered conductive disturbances, which are more frequent in the TIA patients. In the TIA group the significantly more frequent features are acute onset of the neurological deficit, significantly more frequent normal CT scan images or lacunar infarctions findings. In the minor stroke group the significantly more frequent features are the subacute onset, more frequent CT findings of vascular encephalopathy or CT scans revealing one large ischaemic zone. CONCLUSIONS: The differences may be explained with the dominant pathogenetic mechanisms in each of the conditions: microembolisation of extracranial vascular origin in TIA and local thrombosis or cardioembolisation in minor stroke. Previous vascular damage in minor stroke patients is more evident. PMID- 17408075 TI - A pharmacoeconomic comparison of monotherapy with Tegretol, Finlepsin and Trileptal (preliminary data). AB - AIM: Pharmacoeconomics comprises a system of relatively new methods that inform policy makers about the costs and benefits of different therapies so that limited health care resources may be allocated efficiently. The purpose of this study was to perform a pharmacoeconomic comparison of three of the most widely used antiepileptic drugs in Bulgaria: carbamazepine (two different products: Tegretol and Finlepsin) and oxcarbazepine (Trileptal). PATIENTS AND METHODS: The inclusion criteria for entering the study were: women or men of age with newly detected and clinically diagnosed epilepsy,monotherapy treatment with the studied drugs, ability to keep personal records. The follow-up visits were conducted in three month periods, each visit including a thorough evaluation of effectiveness, costs and quality of life. Effectiveness was assessed as seizure reduction percentage and time till new seizure. Costs included direct and indirect medical and nonmedical costs, related to epilepsy or its treatment. The quality of life of the patient was assessed with Quality of Life in Epilepsy Inventory 31 (QOLIE)- an evaluation instrument accepted for use in clinical practice. The statistical methods used included: descriptive analysis, Pearson correlation and Kaplan-Meyer analyses. RESULTS: The mean seizure reduction was 86.05% for Finlepsin, 83.31% for Tegretol and 66.67% for Trileptal. There were no seizure-free patients on Trileptal, while almost 60% of Tegretol and nearly 50% in Finlepsin patients remained seizure-free for a period of one year. CONCLUSIONS: The quality of life of all patients was high, indicating good efficacy and safety as assessed by patients. There was a significant difference in terms of costs in the Trileptal group compared with the other two groups --it incurred higher annual costs as well as higher cost-per-QALY value. PMID- 17408076 TI - Therapeutic algorithm in cases with congenital diaphragmatic hernia of the Bochdalek type. AB - The PURPOSE of the present study was to establish the treatment results of the applied approach for preoperative stabilization through mechanical ventilation and delayed surgical intervention as opposed to emergency surgical treatment in children with congenital diaphragmatic hernia. PATIENTS AND METHODS: Twenty-seven children have been treated (nine girls and 18 boys, twenty-four of them- newborns) over a ten-year period. The criteria for successful stabilization are arterial saturation of SaO2 90%, PaO2 = 90-100 mm Hg, PaCO2 40 mm Hg, pH 7.35. RESULTS: Twenty-five of the children were with left-sided hernia (92.59%) and only two (7.41%) with right-sided hernia. The stabilization period ranged from 1 to 4 days, mean 1.48 +/- 0.18 (Sx = 0.81). Preoperatively, in eighteen newborns we used conventional mechanical ventilation; in one child only we used high frequency oscillating ventilation. In three children we failed to achieve stabilization and after worsening of their condition they died before they could be operated. In all children, postoperative mechanical ventilation was applied from 3 to 16 days, mean 7 +/- 0.65 (Sx = 3.16). The mortality was 59.26%. CONCLUSIONS: The approach used for delayed surgical treatment has significant advantages compared to the urgent correction of the congenital diaphragmatic hernia. It provides opportunities for stabilization of the gas exchange and pulmonary circulation but still mortality remains high. New therapeutic techniques such as antenatal intervention or pulmonary transplantation are probably needed to save the lives of such patients. PMID- 17408077 TI - Bcl-2/Bax ratio, mitochondrial membranes and aerobic enzyme activity in cardiomyocytes of rats after submaximal training. AB - INTRODUCTION: Mitochondria are an active and continuous source of reactive oxygen species (ROS) during respiration. The ROS increased production during endurance training is a result of an augmented electron transport through the respiratory chains, making in this way the mitochondria a potential target for oxidative damage. The Bcl-2 protein family plays a central role in the transition of apoptotic signals towards the mitochondria in stress-induced apoptosis. AIM: The present work studied the effect of endurance training on the expression of the apoptotic proteins Bcl-2 and Bax in rat cardiomyocytes, as well as the concomitant changes in the ultrastructure of the mitochondria and activity of some enzymes residing there. MATERIAL AND METHODS: Two groups of male Wistar rats were used. One was the control and the other was trained on treadmill with submaximal loading for eight weeks. At the end of the trial, samples of the myocardium of all the experimental animals were obtained. Immunohistochemical reactions for Bcl-2 and Bax and enzymehistochemical reactions for succinate dehydrogenase and NADH2-cytochrome C-reductase were done. The results were analyzed using specialized software. Transmission electron microscopical study was carried out too. RESULTS: In the myocardium of the trained animals the expression of Bcl-2 and Bcl-2/Bax ratio were significantly higher compared to the controls. The mitochondria had intact outer and inner membranes, with no signs of swelling. Mitochondria with denser packed cristae were found predominantly. No significant differences were found in the activity of the investigated enzymes in the cardiomyocytes of the animals from both groups. CONCLUSIONS: In the myocardium of the experimental animals endurance training for eight weeks does not lead to activation of apoptotic processes via the mitochondrial pathway. This type of exercise training could be used for cardioprotection in order to elevate apoptotic threshold of cardiomyocytes. PMID- 17408078 TI - Protective effect of Aronia melanocarpa fruit juice pretreatment in a model of carbon tetrachloride-induced hepatotoxicity in rats. AB - The main active ingredients of Aronia melanocarpa fruit juice (AMFJ) are phenolic substances, mainly flavonoids from the anthocyanin subclass. AIM: The aim of the present study was to investigate the effect of AMFJ applied as pretreatment in a model of carbon tetrachloride (CCl4)-induced hepatotoxicity in rats. MATERIAL AND METHODS: AMFJ was given orally to rats for 2 days at doses of 5, 10 and 20 ml/kg either alone or as pretreatment before the oral application of CCl4 (0.2 ml/kg, 2 days). The plasma activities of aspartate transaminase (AST) and alanine transaminase (ALT) were measured as markers of the liver cell damage. The levels of malondialdehyde (MDA), a lipid peroxidation marker, were determined in rat liver and plasma. RESULTS: Administration of CCl4 caused an elevation of plasma AST and ALT activities. It also induced an elevation of MDA levels in rat liver and plasma. AMFJ applied alone in the tested doses did not cause any significant changes in the measured enzyme activities and in MDA levels. AMFJ applied as pretreatment prevented the CCl4-induced increase of AST and ALT activities, and also prevented the elevation of plasma and liver MDA levels. CONCLUSIONS: AMFJ showed a protective effect in a model of CCl4-induced hepatotoxicity in rats. This effect might be due to the antioxidant activity of its active ingredients. PMID- 17408079 TI - A study of post-operative sensitivity in class I and class II restorations with self-etching adhesive and nanofilled composite. AB - AIM: The aim of the study was to determine the type, intensity and duration of post operative sensitivity (POS) in class I and II cavity restorations with self etching adhesive and nanofilled composite. PATIENTS AND METHODS: The clinical study included 34 patients, who received a total of 76 restorations by undergraduate dental students. The restorations were made in medium and deep cavities. Self-etching adhesive Adper Promp L-Pop and nanocomposite Filtek Supreme (3MESPE Dental products USA) were placed. The type, the magnitude and the duration of postoperative sensitivity were examined on days 1, 3, 5, 7, 14 and 30. Intensity of sensitivity perceptions were recorded on a visual analogue scale using the method of self-observation. Parametric and non-parametric analyses were employed at a significance level of P < 0.05. RESULTS: Pressure-dependent POS was present in most of the cases (15.78% +/- 4.209), followed by discomfort (9.21% +/ 3.340) and pressure- and cold-induced pain (1.33% +/- 1.147). A statistically significant difference was found between days 1 and 3, and between days 5 and 30, as well as between days 7 and 30 (Wilcoxon Signed Ranks Test, P < 0.05). Rapidly disappearing POS (within 1 week) had the highest percentage: 75% +/- 9.934. CONCLUSIONS: Following restoration of class I and II cavities with self-etching adhesive and nanofilled composite POS was established in 26.3% of the cases. PMID- 17408080 TI - Raw materials. AB - Industrial fermentations need raw materials that fulfill the requirements of the organism (suitable carbon and nitrogen source, minerals and specific nutrients) and that are available in a high quantity and quality. This contribution gives a comprehensive overview, including the new trends and progress of recent years. The use of feedstock based on several raw materials such as sugar, starch, inulin and lignocellulose is discussed. Biomass-based raw materials are by far the most applied feedstocks for fermentation. However, there are also raw materials for fermentations derived from the petrochemical industry. These substrates are especially hydrocarbons, alcohols and carboxylic acids. Some applications are given in this chapter. PMID- 17408081 TI - Screening systems. AB - Enzyme screening technology has undergone massive developments in recent years, particularly in the area of high-throughput screening and microarray methods. Screening consists of testing each sample of a sample library individually for the targeted reaction. This requires enzyme assays that accurately test relevant parameters of the reaction, such as catalytic turnover with a given substrate and selectivity parameters such as enantio- and regioselectivity. Enzyme assays also play an important role outside of enzyme screening, in particular for drug screening, medical diagnostics, and in the area of cellular and tissue imaging. In the 1990s, methods for high-throughput screening of enzyme activities were perceived as a critical bottleneck. As illustrated partly in this chapter, a large repertoire of efficient screening strategies are available today that allow testing of almost any reaction with high-throughput. PMID- 17408082 TI - Industrial enzymes. PMID- 17408083 TI - Building blocks. AB - This contribution illustrates the versatility of fundamental approaches in industrial biotransformations. The applicability of biotechnology in organic synthesis on an industrial scale is discussed, followed by an overview of historical development and future progress. This chapter depicts three different approaches for the use of biocatalysts in production processes: non-chiral synthesis, asymmetric synthesis, and racemic and dynamic resolution. Applications for whole cells and isolated enzymes as catalysts are introduced. Finally, critical but optimistic conclusions are given. PMID- 17408084 TI - Biorefineries--multi product processes. AB - The development of biorefineries represents the key for access to an integrated production of food, feed, chemicals, materials, goods, and fuels of the future [1]. Biorefineries combine the necessary technologies of the biogenic raw materials with those of intermediates and final products. The main focus is directed at the precursors carbohydrates, lignin, oils, and proteins and the combination between biotechnological and chemical conversion of substances. Currently the lignocellulosic feedstock biorefinery, green biorefinery, whole corn biorefinery, and the so-called two-platform concept are favored in research, development, and industrial implementation. PMID- 17408085 TI - Enabling technologies: fermentation and downstream processing. AB - Efficient parallel tools for bioprocess design, consequent application of the concepts for metabolic process analysis as well as innovative downstream processing techniques are enabling technologies for new industrial bioprocesses from an engineering point of view. Basic principles, state-of-the-art techniques and cutting-edge technologies are briefly reviewed. Emphasis is on parallel bioreactors for bioprocess design, biochemical systems characterization and metabolic control analysis, as well as on preparative chromatography, affinity filtration and protein crystallization on a process scale. PMID- 17408086 TI - Future aspects of bioprocess monitoring. AB - Nature has the impressive ability to efficiently and precisely control biological processes by applying highly evolved principles and using minimal space and relatively simple building blocks. The challenge is to transfer these principles into technically applicable and precise analytical systems that can be used for many applications. This article summarizes some of the new approaches in sensor technology and control strategies for different bioprocesses such as fermentations, biotransformations, and downstream processes. It focuses on bio- and chemosensors, optical sensors, DNA and protein chip technology, software sensors, and modern aspects of data evaluation for improved process monitoring and control. PMID- 17408087 TI - Deaths: leading causes for 2003. AB - OBJECTIVES: This report presents final 2003 data on the 10 leading causes of death in the United States by age, race, sex, and Hispanic origin. Leading causes of infant, neonatal, and postneonatal death are also presented. This report supplements the annual report of final mortality statistics. METHODS: Data in this report are based on information from all death certificates filed in the 50 states and the District of Columbia in 2003. Causes of death classified by the International Classification of Diseases, Tenth Revision (ICD-10) are ranked according to the number of deaths assigned to rankable causes. RESULTS: In 2003, the 10 leading causes of death were (in rank order): Diseases of heart; Malignant neoplasms; Cerebrovascular diseases; Chronic lower respiratory diseases; Accidents (unintentional injuries); Diabetes mellitus; Influenza and pneumonia; Alzheimer's disease; Nephritis, nephrotic syndrome and nephrosis; and Septicemia and accounted for about 78 percent of all deaths occurring in the United States. Differences in the ranking are evident by age, sex, race, and Hispanic origin. Leading causes of infant death for 2003 were (in rank order): Congenital malformations, deformations and chromosomal abnormalities; Disorders related to short gestation and low birth weight, not elsewhere classified; Sudden infant death syndrome; Newborn affected by maternal complications of pregnancy; Newborn affected by complications of placenta, cord and membranes; Accidents (unintentional injuries); Respiratory distress of newborn; Bacterial sepsis of newborn; Neonatal hemorrhage; and Diseases of the circulatory system. Important variation in the leading causes of infant death is noted for the neonatal and postneonatal periods. PMID- 17408088 TI - [Receptors of the serpentine type and heterotrimeric G-proteins of the yeasts: structural-functional organization and molecular action mechanisms]. AB - The signal systems of the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, coupled to heterotrimeric G-proteins and sensitive to pheromones and alimentary molecules, are prototypes of hormonal signal systems of the higher vertebrate animals and are widely used in studies on molecular mechanisms of their functioning. This review summarizes and analyzes data on structural functional organization of the first two components of these systems - receptors of the serpentine type and heterotrimeric G-proteins; mechanisms of functional coupling of receptors and G-proteins both between each other and to other signal proteins are discussed. It has been shown that at the early stages of evolution of signal systems, at the yeast level, various models of transduction of signals into the cell were tested; many of them differ essentially from the classic model of the three-component, G-protein-coupled signal system of the higher vertebrates. PMID- 17408089 TI - [Development of respiratory function in perinatal ontogenesis]. AB - In development of respiratory function in rats, mice, and other representatives of placental animals there exists the general plan of formation of rhythm: from single contraction of respiratory musculature to formation of bursts and complexes alternating periodically with pauses and apnea intervals and subsequent rhythm stabilization. These peculiarities are closely connected with the states of sleep and consciousness. A concept is put forward about a certain sequence of functional maturation and ways of regulation of activity of the respiratory rhythm central pacemaker. At the first stage the autogenic rhythmical activity is determined by pacemaker properties of a part of neurons of the medulla rostral ventrolateral part. It is not ruled out that the first respiratory discharges in spinal cord ventral roots might have been a manifestation of the nervous network rhythmogenic properties. The direct sensitivity of central neurons to chemical composition if the medium and to some neutomodulators serves as the first regulatory mechanism. Somewhat later, inhibitory control is established from supramedullary structures, with an increase of role of peripheral receptors in regulation of respiration. PMID- 17408090 TI - [Activation of cardiac contractivity of the edible snail H. pomatia under effect of thrombin. Study of role of cAMP]. AB - Thrombin acts on mammalian cells through specific, the so-called protease activated receptors (PARs). The thrombin action is mediated via three out of four known types of these receptors PAR(1, 3, 4). Mammalian thrombin receptors, apart from performance of other functions, control cardiac and vascular contractility. It is not known whether receptors of such kind exist in invertebrate animals. In the present work we have showed for the first time that thrombin in the concentration range of 0.01-1 units/ml increases amplitude of contractions of the isolated heart ventricle of the edible snail Helix pomatia. Its effect is reproduced by peptide ligands of receptors PAR1 and PAR4 that have sequences Ser Phe-Leu-Leu-Arg-Asn (SFLLRN) and Glu-Tyr-Pro-Gly-Lys-Phe (QYPGKF), respectively. A potent activati of cardiac contractivity of H. pomatia is serotonin. A comparative study of mechanisms of action of serotonin and thrombin on the edible snail heart was carried out. cAMP participates in transduction of signal from serotonin receptors. On the membrane preparation from the H. pomatia heart, it was shown that thrombin and peptide ligands PAR(1, 4), unlike serotonin, did not increase adenylyl cyclase activity. Thus, mechanism of activation of cardiac contractivity of H. pomatia by thrombin differs from the action mechanism of serotonin. It is suggested that molluscs have receptors homologous to protease activated mammalian receptors. PMID- 17408091 TI - [Peculiarities of dopamine receptors on the membrane of spinal cord multipolar neurons of the brook lamprey Lampetra planeri]. AB - On isolated multiporal neurons of spinal cord of amniocoete larva of the brook lamprey Lampetra planeri, by the patch-clamp method in configuration "the whole cell", a modulating effect of dopamine on potential-activated Na+ currents was studied. Application of dopamine (10 microM) was shown to produce a complex action on the sodium current amplitude. In some cases a decrease of the amplitude, on average, by 13.5 +/- 2.2% was found, while in others--an increase, on average, by 8.6 +/- 6.1%. The modulation dopamine effect was not accompanied by any changes either of the threshold of the current appearance or of resistance of neuronal cell membranes. Pharmacological analysis with use of dopamine agonist has shown that the agonist of D1-receptors (-)-SKF-38393 (10 microM) decreases the Na+ current amplitude, whereas the agonist of D2-receptors (-)-quinpirole (10 microM) can produce in different cells both an increase, by 30.7 +/- 17.0 %, and a decrease, by 13.2 +/- 3.1%, of the Na+ current amplitude. The obtained data indicate the existence of D1- and D2-receptors on the membrane of multipolar spinal neurons of the amniocoete larva of the brook lamprey. Study of action of antagonists has shown that the antagonist of D1-receptors (+)-SCH-23390 (10 microM) does not affect action of the agonist of D1-receptors (-)-SKF-38393 (10 microM); the antagonist of D2-receptors (-)-sulpiride (10 microM) blocks completely effects both of the agonist of D1-receptors (-)-SKF-38393 (10 microM) and of the agonist of D2-receptors (-)-quinpirole (10 microM). The antagonist of D1-receptors (+)-SCH-23390 (10 microM) produced no effect on action of the agonist of D1-receptors (-)-SKF-38393 (10 microM). The obtained data indicate peculiarities of dopamine receptors of Cyclostomata as compared with those in mammals. PMID- 17408092 TI - [Electroencephalographic analysis of manifestations of latent forms of paroxysmal syndrome in the wakefulness-sleep cycle in rats]. AB - An electroencephalographic study of the brain activity in the wakefulness-sleep cycle was carried out on rats of Krushinskii-Molodkina line (KM) with hereditary predisposition to audiogenic convulsions and on Wistar rats that were insensitive to the convulsiogenic sound effect, but with epileptiform manifestations appearing on the background of cadmium intoxication and administration of kainic acid into the caudate nucleus head. There were revealed several EEG patterns whose presence was an indicator of formation of disorders of the CNS activity of the paroxysmal character in the animals. It has been established that in the phase of the rat rapid-wave sleep, a high representation of episodes with predominance of a-diapason EEG oscillations can be considered a specific non paroxysmal abnormality due to the presence of convulsive syndrome in these animals. It was shown the long steady decrease of sensitivity of KM rats to the convulsiogenic sound effect, which appeared after multiple audiogenic generalized tonicoclonic convulsive attacks, correlated with a decrease of the degree of theta-diapason oscillations and with an increase of representation of alpha diapason waves on EEG in the state of the animal quiet consciousness. A role of disintegration in activity of the ascending activating brain systems in the animal and human paroxysmal syndromes is discussed. PMID- 17408093 TI - [Sex-dependent differences in parameters of a long-term pain caused by inflammatory focus in prenatally stressed newborn rats]. AB - In experiments on the 7-day-old female and male Long-Evans rat pups, for the first time, there was studied effect of prenatal (immobilization) stress on dynamics of nociceptive behavioral response caused by an inflammatory focus. The nociceptive sensitivity was evaluated for 1 h by the number of 7-day-old organized at the spinal level in response to injection of formalin (10%, 10 microl) to the posterior leg sole. Control rat pups were not submitted to any prenatal stress; in these animals the response in the formalin test was found to be represented by one phase. It the prenatally stressed rat pups the studied patterns were organized into two phases characteristic of the definitive type of response. At the period between them (during interphase), the nociceptive behavior was absent. At the second, tonic phase the number of flexes+shakes in the prenatally stressed males was statistically significantly higher than in the prenatally stressed females, which indicates a sensitization of the neurons involved in the tonic pain chains in male individuals. Thus, the data obtained on prenatally stressed animals confirm the previous data about immaturity of the mechanisms mediating the second phase of response in the formalin test in the 7 day-old rat pups. An important fact is revealed which indicates that in the prenatally stressed rat pups of the same age the second phase of response is already obvious. Mechanisms underlying the behavioral response caused by the inflammatory focus in the formalin test in the number flexes + shakes old stressed rat pups are characterized by sexual dimorphism: the pain sensitivity in males at the second phase of response is statistically significantly higher than in females. PMID- 17408094 TI - [Monoamine- and peptide-containing elements in the nemertine digestive tract]. AB - Detailed comparative-histological data on localization and morphological peculiarities of catecholamine-, serotonin-, neurotensin- and FMRFamide containing elements in the nemertine digestive system are presented for the first time. Using fluorescent histochemistry and immunohistochemistry, pharynx, esophagus, and midgut were studied in five species from three genera of the White Sea nemertines. Described in various nemertine species are intra- and subepithelial cells of the open and closed type, containing biologically active substances. Their processes are distributed in basi- and subepithelial nerve plexuses and can make contact with lumen of the digestive tract. Species specificity in localization of the cells containing certain biologically active substances is noticed both along the length of the tract and with respect to its epithelial layer. Peculiarities and common regularities in distribution are discussed, as well as a possible function of monoamine- and peptide-containing elements in the digestive tract of the studied nemertines and other invertebrates. PMID- 17408095 TI - [Periventircular and central nuclei of the diencephalon pretectal area of the sturgeons]. AB - Cytoarchitectonics of periventricular and central nuclei of the pretectal area was studied in four species of the sturgeons: the great sturgeon Huso huso, L., the Russian sturgeon Acipenser guldenstadti persicus n. kurensis, Belyaeff, the starred sturgeon Ac. stellatus, Pall., and the barbel sturgeon Ac. nudiventris, Lov.; this pretectum part has a similar structure. Study of these parts of the pretectal area was carried out by methods of Nissl and Bielshowskii modified by Viktorov. In this part of the pretectal area, nine nuclear structures were described, eight of them--nuclear; these are ventral periventricular pretectal nucleus and its dorsal component, dorsal periventricular pretectal nucleus, nucleus of medial longitudinal bundle, subcomissural organ, medial and lateral intercalate nuclei, and central and posterior pretectal nuclei. The main attention has been paid to the issue of the evolutional progression of this part of the pretectal area in the sturgeons as compared with other Actinopterygii. PMID- 17408096 TI - [Outer nuclei of the diencephalon pretectal area of the sturgeons]. AB - Cytoarchitectonics of outer nuclei of the pretectal area was studied in four species of the sturgeons: the great sturgeon Huso huso, L., the Russian sturgeon Acipenser guldenstadti persicus n. kurensis, Belyaeff, the starred sturgeon Ac. stellatus, Pall., and the barbel sturgeon Ac. nudiventris, Lov. Morphological organization of these structures was studied by methods of Nissl and Bielshowskii modified by Viktorov. In this part of the pretectal area, five nuclear structures were described: parvicellular and magnocellular outer pretectal nuclei, lamellar pretectal nucleus, and dorsal pretectal nucleus, as well as accessory optic nucleus. Comparison of the obtained results with literature data has revealed a sufficiently high level of differentiation of this pretectum part of the sturgeons. Besides, a general conclusion is presented on organization of their entire pretectal area. PMID- 17408097 TI - [The level of metabolic activity (cytochrome oxidase) as an index of functional significance of tectofugal and thalamofugal channels of the reptilian visual system]. AB - Using histochemical determination of activity of the mitochondrial oxidative enzyme cytochrome oxidase (CO) in brain structures, metabolic activity both in turtles and in lizards has been shown to be higher in centers of the tectofugal channel (the tectal stratum griseum centrale, SGC; nucleus pretectalis ventralis, Ptv; thalamic nucleus rotundus, Rot; telencephalic visual area of the anterior dorsal ventricular ridge, Advr) than in the thalamofugal channel centers (the thalamic nucleus geniculatus lateralis pars dorsalis, GLd; cortex dorsolateralis, Cxdl; and pallial thickening, Path) of the visual system. Some interspecies differences in distribution of the CO activity in the tectal, thalamic, and telencephalic visual centers between terrestrial and pond turtles and lizards were revealed. The obtained data confirm the idea on the dominating role of the tectofugal channel over the thalamofugal channel of the visual system in information processing and organization of the day-to-day behavior of reptiles. PMID- 17408098 TI - [Transmural gradient of recovery of excitability of the left ventricle myocardium of the chicken Gallus domesticus]. PMID- 17408099 TI - [Preservation of level of parasympathetic effect on human heart under conditions of acute hypoxia as an index of high hypoxic resistance. Wavelet-analysis]. PMID- 17408100 TI - [Search for evolutionary ancient, relict regulatory peptides]. AB - Among numerous regulatory peptide (RP) it is possible to presumably indicate the relict, evolutionary ancient RP. They combine three features: formation from non specialized proteins-precursors, a comparatively high resistance to action of proteases in the organism media, and maximal simplicity of their structure. The examples of them are glyprolines--a recently identified RP family, as well as tuftsin. Several other praline-containing RP in terminal sites also seem to belong to the evolutionary ancient RP. The proposed approach to studies on the RP evolution is additional to those used traditionally in this problem. PMID- 17408101 TI - [Risk factors for birth injuries]. AB - OBJECTIVE: To identify risk factors associated with birth trauma. SETTING: Servicio de Neonatologia, Hospital General "Dr. Manuel Gea Gonzalez", Secretaria de Salud. DESIGN: Case-control, prolective study. PATIENTS: There were 129 cases and 134 controls. MEASURES: We recorded the following variables: a) maternal and delivery: age, weight, height, prenatal care, pre-existing disease or gestational disease, mode of delivery, anesthetic management during labor, use of external maneuvers or forceps; b) newborn: birth weight, gestational age, academic degree of attendant physician at delivery, and type of birth injury. RESULTS: The independent risk factors associated to birth injury were: for ecchymoses; general anesthesia (OR 13.7, 95% CI = 3 - 62.6), breech presentation (OR 6.4, 95% IC 95% = 1.4 - 27.9) and gestational age < or = 32 weeks (OR 6.4, 95% CI = 1.3 - 31.1); for lacerations, vaginal dystocic delivery or cesarean section (OR 19, 95% CI = 4.4 - 81.1) and use of external maneuvers (OR 5.6, 95% CI = 1.5 - 21.6); for cephalhematoma maternal height < or = 1.54 m (OR 7.4, 95% CI = 2.3 - 23.7) and external maneuvers (OR 7.2, 95% CI = 2.3 - 23.7); for caput succedaneum, external maneuvers (OR 3.4, 95% CI = 1.5-7.7) and maternal age < or = 19 or > or = 36 years (OR 3.0, 95% CI = 1.4 - 6.4). CONCLUSIONS: Risk factors associated with birth injuries identified in this study involved maternal conditions, neonatal conditions and mechanism of delivery. PMID- 17408102 TI - [Thalidomide-associated bradycardia in patients with hematologic diseases: a single institution experience]. AB - Thalidomide, an immunomodulatory and antiangiogenic agent, is useful in the treatment of some hematologic and oncologic diseases. Up to 6.8% of thalidomide treated patients present bradycardia. Herein the incidence of thalidomide associated bradycardia in patients with hematologic diseases treated in a single institution is reported. In a 34-month period, 33 patients with different hematologic diseases (multiple myeloma [MM], 20; myelodysplastic syndrome, eight; Waldenstrom macroglobulinemia, two; non-Hodgkin's lymphoma, two; malignant histiocytosis, one) were treated with thalidomide. Of them, five (15.1%) had bradycardia, all with MM. Bradycardia was detected with a daily thalidomide dose ranging from 100 to 300 mg and the time patients received thalidomide before cardiac event went from one to 18 months. In all affected cases the electrocardiogram showed sinus bradycardia with cardiac frequency between 32 to 48 beats per minute. Time to normal cardiac beat recovery ranged from 12 to 21 days after thalidomide discontinuation. There were no fatalities due to thalidomide-associated bradycardia. It is concluded that: a) thalidomide associated bradycardia was detected only in patients with MM, b) herein the incidence of bradycardia was higher as compared with other series, and c) in patients with MM thalidomide therapy must be prescribed with caution particularly in those with cardiovascular diseases of any etiology. PMID- 17408103 TI - [Validation of a clinimetric scale for the diagnosis for depression in patients with diabetes mellitus type 2, in primary health care]. AB - BACKGROUND: The prevalence of depression in patients with type 2 diabetes mellitus (DM2) is of up to 49.3% in primary care clinics. Nevertheless, medical doctors only recognize only 30% of these cases. Depression is associated with poor glycemic control, increase of diabetes complications, deterioration in patient's quality of life, and increase in demand and resources to provide care. The objective was to design and validate a clinimetric scale for the diagnosis of depression (CSDD in patients with DM2, in primary care units. PATIENTS AND METHODS: The study was conducted on 528 DM2 patients in Family Medicine Unit No. 10 of the Instituto Mexicano del Seguro Social (Mexican Social Security Institute), during 2003. A diagnostic test design was employed, with the golden standard consisting of the composite international diagnostic interview. Samples were constructed around consecutive cases. Depression and its degrees were the dependent variables. Absolute and relative frequencies were calculated, along with the Kappa index, sensibility, specificity, positive predictive values (PPV) and negative predictive values (NPV) and ROC curves. RESULTS: The CSDD presented a concordance between observers of 0.7739. The best cut-off point in the ROC curves for diagnosis of depression was 6, which obtained a sensibility of 95.3%, a specificity of 96.8%, a PPV of 92.2%, and a NPV of 98.1%. CONCLUSIONS: The CSDD is a consistent and valid instrument and easy to use for the diagnosis of depression in patients with DM2 in primary care clinic. PMID- 17408104 TI - [Kikuchi-Fujimoto disease (histiocytic necrotizing lymphadenopathy) clinicopathologic and immunohistochemical study of 14 cases and its differential diagnosis with other reactive and neoplastic necrotizing lymphadenopathies]. AB - Kikuchi-Fujimoto disease also known as histiocytic necrotizing lymphadenopaty (HNL) is a rare entity, originally described in Japanese population, although currently it has been described all over the world. It is more frequent in young women and it is usually located in cervical lymph nodes. We report 14 cases of HNL in Mexican population, their clinicopathological and immunohistochemical study as well as a comparative study with other necrotizing lymphadenopaties due to B or T-cell lymphomas, tuberculosis, Epstein Barr virus infection, and non specific necrosis. In our study we found that there was more expression of the immunomarkers CD68, MPO, CD123 and antikerat in OSCAR in the cases of HNL in contrast with the lesser or even null expression of the same markers in the necrotized lymph nodes of the comparative study group. PMID- 17408105 TI - [Co-morbidity and mortality during the first year of life in children with jejunoileal atresia]. AB - OBJECTIVE: To identify the co-morbidity and mortality during the first year of life in children with jejunoileal atresia. DESIGN: Descriptive, comparative cohort. SITE: Hospital de Pediatria, Centro Medico Nacional Siglo XXI, Instituto Mexicano del Seguro Social. POPULATION: Seventy children with jejunoileal atresia. MEASURES: Following variables were registered: gestational age, birth weight, sex, type and site of atresia, other congenital defects, surgical management, number of surgeries, duration of fasting, re-hospitalizations, weight and height at 6 months and one year of life, co-morbidity and mortality. RESULTS: Intestinal atresia types I and II were present in 27% of the patients respectively and type IIIa in 24%. Co-morbidity in the neonatal period was 64.2%, being the most common conditions sepsis (47.1%) and anastomotic stricture (18.5%). During the first year of life co-morbidity was 51.4%, presented as functional intestinal obstruction (21.4%). Mortality was 11.4% (n = 8), the main causes of death were sepsis (n = 6) and liver failure (n = 2). The mortality rate according to the type of atresia was 44.4% for type IV, 33.3% for type IIIb and 1.3% for type I. CONCLUSIONS: Co-morbidity during the first year of life in children with jejunoileal atresia is high, and related to infections and anastomosis inherent problems. Mortality is higher for intestinal atresia type IV. PMID- 17408106 TI - The Janus Kinase 2 (JAK2) V617F mutation in hematological malignancies in Mexico. AB - A new mutation (V617F) affecting the JAK2 gene has been recently described as acquired in patients with myeloproliferative disorders and other myeloid malignancies. Using an amplification refractory mutation system, we investigated this mutation in 70 Mexican mestizo patients with hematological malignancies: 28 cases of acute lymphoblastic leukemia, 17 cases of Ph1-positive chronic myelogenous leukemia, 8 patients with acute myelogenous leukemia, 6 patients with chronic lymphocytic leukemia, 6 patients with polycythemia vera (PV), two patients with essential thrombocythemia (ET), one patient with hypereosinophilic syndrome one patient with primary myelofibrosis (MF) and one patient with chronic myelomonocytic leukemia. The mutation was identified in 4 of 6 patients with PV, in one of 2 patients with ET and in the patient with MF. Our data add to the observation that the JAK2 V617F mutation seems to be rather uncommon in myeloid malignancies other than the classic BCR/ABL negative MPD. PMID- 17408107 TI - [The effect of MTHFR polymorphisms, pregnancy and first intercourse on cervical cancer in a population from the Northeastern Mexico]. AB - OBJECTIVE: To investigate the possible association among MTHFR polymorfhisms, environmental factors and cervical cancer (CC) in the Mexican population. METHODS: Seventy patients with CC and 89 control women were questioned about clinical data and their 677 and 1298 genotypes of MTHFR gene were analized. RESULTS: Multipregnancies (0-2 vs. > or = 3, OR 2.1), an early age of first intercourse (IVS) (17 < or = vs. > or = 18 years, OR 4.3) or both factors (OR 3.5) were significantly associated with CC. MTHFR 677, 1298 polymorphisms and their combinations were not different between cases and controls. However, a significant association between pregnancies, IVS and MTHFR polymorphisms (presence of 1298C allele or 677TT genotype) was observed. The 1298C allele plus multipregnancies and IVS < or = 17 years, or both factors, increased 4.3, 5.3, and 11.8 times the risk for CC, respectively, while 677TT genotype changed the risk 2.0, 1.9, and 4.2 times, respectively. CONCLUSION: The 1298C allele increases the risk of CC strongly in women with multipregnancies and early age of IVS, while 677TT genotype has a lower risk without becoming a protection factor. PMID- 17408108 TI - [Chlorination and bacterial contamination. Hospitals with troubled waters]. AB - OBJECTIVE: To know the quality of the water from hospitals and the risks that poor chlorination implies. METHODS: We analyzed 90 water specimens from 15 hospitals (9 from the private and 6 from the public sector). Specimens were obtained from three areas (hospitalization ward, operating room, and kitchen) in two visits. RESULTS: By microbiologic analysis, we found 30 (33.3%) contaminated specimens. By chemical analysis, we found only 49 (54.5%) specimens with adequate chlorination (> or = 1 mg/L). Of the 30 contaminated specimens, only 7 (23.3%) had adequate chlorination. On the other hand, of the 60 specimens without contamination, 42 (79%) had adequate chlorination (chi2 = 17.561, p < 0.001). The significant difference was sustained when the criterion for appropriate chlorination was established at > or = 0.5 mg/L. CONCLUSIONS: Hospitals require guidelines for water chlorination and testing to maintain it at > or = 0.5 mg/L in every point of use. PMID- 17408109 TI - [Ultrastructural study of platelets patients with paroxysmal nocturnal hemoglobinuria]. AB - INTRODUCTION: Paroxysmal nocturnal hemoglobinuria (PNH) is a hemolytic anemia characterized by intravascular hemolysis, cytopenias and venous thrombosis. Previous studies in patients with PNH have shown platelet abnormalities; however, their association with the clinical development of the sickness has not still been determined. MATERIAL AND METHODS: In this study, we compared the morphology and distribution pattern of actin, myosin, tubulin and p-selectin in resting and activated platelets from 22 PNH patients and healthy donors by transmission electron microscopy and immunofluorescence. RESULTS: The PNH platelet ultrastructure of resting and activated with different agonists (ADP, collagen and thrombin) showed morphological changes which suggested the presence of circulating platelets. The developed structures during the adhesion process (filopodia and lamellipodia formation), as well as the pattern distribution of actin, myosin, tubulin and p-selectin in PNH platelets were not modified in relation to control platelets. CONCLUSION: Morphological changes in resting platelets were related with p-selectin expression suggesting its determination as thrombosis indicator. PMID- 17408110 TI - [Glycemic, insulinemic index, glycemic load of soy beverage with low and high content of carbohydrates]. AB - Consumption of soy has increased in Western countries due to the benefits on health and the attitude of the people to consume natural products as alternative to the use of pharmacological therapies. However, there is no evidence whether the consumption of 25 g of soy protein as recommended by the Food and Drug Administration has some effect on glucose absorption and consequently on insulin secretion. The aim of the present study was to determine glycemic index (GI), insulinemic index (InIn), and glycemic load (GL) of several soy beverages containing low or high concentration of carbohydrates, and compare them with other foods such as peanuts, whole milk, soluble fiber and a mixed meal on GI and InIn. The results showed that soy beverages had low or moderate GI, depending of the presence of other compounds like carbohydrates and fiber. Consumption of soy beverages with low concentration of carbohydrates produced the lowest insulin secretion. Therefore, these products can be recommended in obese and diabetic patients. Finally soy beverages should contain low maltodextrins concentration and be added of soluble fiber. PMID- 17408111 TI - [Mesenchymal stem cell; history, biology and clinical application]. AB - In the last years, stem cells have drawn the attention of various sectors of society for many reasons. From the basic point of view, stem cells represent an ideal model to study cell differentiation and self-renewal mechanisms. However, their potential in cell therapy and regenerative medicine has triggered the increasing amount of knowledge in this area. Mesenchymal stem cells belong to the select group of adult stem cells. They have differentiation potential towards mesenchymal tissues such as bone, cartilage, stroma and fat. Recently, both in vivo and in vitro reports have shown a greater plasticity of mesenchymal stem cells, showing not only a mesenchymal cell fate but also those leading to endothelial, nervous and muscular lineages. For these reasons, the study of mesenchymal stem cells has gained great interest and many articles have been published. In the present review, we have presented a global vision of this topic, including its history, biologic features, sources, isolation methods and an overview on their clinical application. PMID- 17408112 TI - [Contributions of pharmacogenomics to enhance the efficiency of randomized clinical trials]. AB - One of the greatest advances of the modern medicine has been the report of the complete sequence of the human genome. This has brought as a consequence an evolution in the design of the clinical research, in special of the randomized clinical trials (RCTs). The pharmacogenomics, a powerful tool for the prediction of pharmacological effects based on the genotype of the studied subjects, promises to be very useful next years for the development of the pharmaceutical industry. With the present integration of the pharmacogenomical methods to the investigation and development of new medicines it may start a new era in the medical prescription producing more individualized therapies, reduction of adverse events in the patients and in addition a faster development of new medicines in a more cost-effective way. Nevertheless new methodological, ethical and social challenges appear that will have to be solved simultaneously, to allow a legal use of the vast information generated by the genetic information. PMID- 17408113 TI - Editorial overview: cardioprotective drugs for myocardial ischemic injury--a therapeutic area at risk. PMID- 17408114 TI - Cardiovascular developments: patent highlights July to December 2006. AB - This overview provides a follow-up to a previously published paper on patent highlights of cardiovascular developments for January to June 2006. A period of six months could not have altered the fundamental preliminaries of the developmental challenges which cardiovascular disorders present to the pharmaceutical and biotechnology industry, and it was not expected that scientific insight would make quantum leaps during this brief interval. What makes such a repeat analysis nevertheless highly meaningful, beyond providing just another set of recent patent references, is the objective of validation- investigating whether the patterns that were observed during the initial review of the first half of 2006 actually represent established modalities of innovative industry efforts in the cardiovascular field. This objective was met by replicating the previous search of Thomson Scientific's Investigational Drugs database, and by using the same categorization of identified intellectual property disclosures. PMID- 17408115 TI - Apolipoprotein AI mimetic peptides: possible new agents for the treatment of atherosclerosis. AB - There is increasing evidence that therapeutic agents for raising HDL would be a useful addition to the current treatment approach for preventing coronary heart disease (CHD), especially considering the fact that therapies for lowering LDLs are not fully adequate for preventing CHD. The recent unraveling of some of the complexities of HDL metabolism has led to the identification of new key proteins involved in HDL metabolism, thus giving new hope and ideas for drug targets. This review focuses on apolipoprotein AI mimetic peptides, which are currently being explored as therapeutic agents for a new treatment strategy known as acute HDL therapy. PMID- 17408116 TI - The pharmacogenetics of antiplatelet drugs. AB - Platelets play a pivotal role in coronary heart disease and atherothrombotic events, and thus antiplatelet therapy is commonly used to treat cardiovascular disease. Unfortunately, the benefits of this approach are limited, and a large proportion of treated patients will suffer a new thrombotic event, either within the first few weeks following stent implantation or in the follow-up period after myocardial infarction. This lack of clinical benefit may be related to a heterogeneous response to antiplatelet drugs among individuals. Recently, the field of pharmacogenetics has tried to identify gene variants in order to explain tile observed heterogeneity in patient responses to drugs. This review explores the genetic influences on antiplatelet drug efficacy by analyzing the modulating role of different genetic polymorphisms on individual responses to antiplatelet therapy. PMID- 17408117 TI - In vivo cardiovascular assays for drug discovery: evolution of the drug-eluting stent. AB - Percutaneous intervention using balloon angioplasty accompanied by stent implantation has become the predominant procedure to treat occlusive coronary and peripheral vascular disease. Unfortunately, restenosis associated with intimal hyperplasia and arterial remodeling at the stented site occurs within 6 months in 20 to 30% of cases. To address this problem, the concept of utilizing a stent as the vehicle to deliver agents locally and limit the overexuberant tissue response related to its placement has been developed. Targeting excess arterial wall smooth muscle cell proliferation, preclinical studies have demonstrated the efficacy of two drugs, paclitaxel and rapamycin, in both in vitro and in vivo animal studies. Early, as well as large, randomized clinical studies using polymer-coated, drug-eluting stents have clearly demonstrated a significant and dramatic efficacy in reducing restenosis rates and improving clinical outcomes compared with the use of the bare stent for revascularization procedures. Despite the low incidence of late thrombosis associated with the rapamycin- and paclitaxel-eluting stents, some concerns remain (such as the need for sustained anticoagulant therapy), providing the impetus for developing coated stents that promote rather than inhibit endothelial healing in order to limit the restenotic response. PMID- 17408118 TI - Phosphodiesterase type 5 inhibitors in the treatment and prevention of high altitude pulmonary edema. AB - The combination of a tendency to affect otherwise fit and healthy individuals, and a characteristically rapid progression to death within hours of the first symptoms, renders high altitude pulmonary edema (HAPE) a particularly devastating illness in travelers to high altitudes. The alveolar edema and ventilation:perfusion mismatch initiate a catastrophic downward spiral of worsening alveolar hypoxia. This reviewo discusses the rationale for the use of phosphodiesterase (PDE)5 inhibitors in HAPE, compares the pharmacokinetic properties of the available agents, and appraises the relevant experimental evidence. Although this class of drugs shows promise in high altitude medicine, further research is necessary to determine the efficacy and safety of PDE5 inhibitors as a treatment for established PMID- 17408119 TI - The detection of myocardial ischemia in end-stage renal disease. AB - The outcome of acute coronary syndromes is usually poor in patients with end stage renal disease (ESRD), and the early detection of myocardial ischemia is therefore essential in this group. However, the diagnosis of ischemina is often problematic in patients with ESRD, as cardiac symptoms and electrocardiography are not reliable predictors in this context. Silent ischemia is common and resting ECG is often abnormal in patients with ESRD. Therefore, alternatives to these predictors have been sought, and this review explores the role of myocardial perfusion imaging, dobutamine stress echocardiography and biochemical markers for use in the diagnosis of ischemia and risk stratification in patients with ESRD. PMID- 17408120 TI - Vascular calcification in chronic kidney failure: role of vitamin D receptor. AB - Chronic kidney disease (CKD) patients encounter an increased risk of cardiovascular disease and mortality compared with healthy individuals, most likely due to the presence of severe atherosclerosis and accelerated vascular calcification. Vascular calcification is an active, regulated process resulting from an imbalance between losses in inhibitory factors and gains in inducing factors present in cells and the blood circulation. However, exactly which inhibitory and inducing factors are involved remains unknown. The vitamin D receptor (VDR) is a nuclear receptor present in over 30 different tissues. Several VDR activators (VDRAs), including paricalcitol and calcitriol, are currently available for the treatment of secondary hyperparathyroidism in patients with CKD. Recent clinical observations demonstrate that VDRA therapy provides survival benefits for CKD patients in the order of paricalcitol > calcitriol > no VDRA therapy, independent of serum parathyroid hormone, phosphorus and calcium levels. The survival benefit of VDRAs seems contradictory to the perception that VDRAs, due to their potential impact of increasing serum phosphorus and calcium, may cause calcification in vessels. A review of the current literature shows that inconsistent data exist regarding the role of VDRAs in vascular calcification. A possible explanation is that the VDR may be involved in regulating several different pathways as an endocrine, paracrine and/or autocrine factor, and different VDRAs may have differential effects on the endocrine versus the paracrine/autocrine aspect. PMID- 17408121 TI - Drug evaluation: PB-127, a novel contrast agent for the detection of myocardial perfusion. AB - Point Biomedical Corp is developing PB-127 (CardioSphere) as an ultrasound imaging agent for assessing myocardial perfusion in patients with coronary artery disease. In January 2006, the company submitted an NDA for the agent; this was accepted for review in February 2006, with a Prescription Drug User Fee Act review and action date of late October 2006. PB-127 is effective for intermittent harmonic power Doppler imaging and may be the first contrast agent to receive FDA approval for myocardial perfusion imaging. PMID- 17408122 TI - Drug evaluation: rivaroxaban, an oral, direct inhibitor of activated factor X. AB - Bayer AG and Ortho-McNeil Pharmaceutical Inc are developing rivaroxaban, an oral Factor Xa inhibitor, for the potential prevention and treatment of thrombosis. In December 2005 and December 2006, phase III trials were initiated for the prevention of venous thromboembolism (VTE) following major orthopedic surgery and in patients with atrial fibrillation, respectively. By January 2007, a phase III trial for the prevention of VTE following total knee replacement surgery had commenced. In November 2006, a phase II trial for the reduction of VTE in patients with acute coronary syndrome was initiated. PMID- 17408123 TI - You, your public health department and the next flu pandemic. PMID- 17408124 TI - No time for practice management resolutions. PMID- 17408125 TI - Radiating courtesy and professionalism. PMID- 17408127 TI - EHRs fix everything--and nine other myths. PMID- 17408128 TI - Helping your patients avoid counterfeit medicines. PMID- 17408129 TI - Information from your family doctor. Avoiding counterfeit drugs. PMID- 17408130 TI - A moonlighting position can help you hone your medical skills while boosting your income. Is moonlighting right for you? PMID- 17408131 TI - A refresher on coding consultations. Here's a guide to the sometimes confusing documentation requirements for consultations. PMID- 17408132 TI - Vaccine administration: making the process more efficient in your practice. PMID- 17408133 TI - Dealing with medical junk mail. PMID- 17408134 TI - [What does saliva represent to the dentist?]. AB - Despite the undoubtedly large impact of saliva on the patient's oral health, little is known about its exact properties and role. More research has been put into serum, when compared to saliva, although that is changing. More and more researchers are interested in the role of certain proteins or electrolytes in an oral disease process. Besides the purely laboratory research, practitioners are also interested in the bacterial colonisation of the oral fluids, especially in assessing a patient's caries risk. The use of bacterial chair-side tests illustrates this aspect of progression in oral health care. For the dental professional, saliva is an important ally in preventing dental caries and mucositis. The lubrication properties and antibacterial/antiviral function of saliva play an important role in the protection of the oral cavity. These aspects may be suppressed in some medically compromised patients. In those cases, the dental professional is faced with a challenge to solve the oral health problems. PMID- 17408135 TI - [The pathology of salivary glands. Tumors of the salivary glands]. AB - The management of benign and malignant neoplasms of the salivary glands requires precise knowledge of tumor histogenesis and classification as well as surgical skills. Pleomorphic adenoma and Whartin's tumor are the most frequent tumors in parotid glands while the probability for malignant tumors is higher in other glands, especially in sublingual and minor salivary glands. Those malignant salivary glands tumors are rare and necessitate multidisciplinar staging and management in close collaboration with the pathologist and the radiation oncologist. PMID- 17408136 TI - [Systemic pathology and the salivary glands]. AB - Various systemic diseases can affect salivary glands. After a short anatomical and semiological recall, we describe the pathologies according to their possible salivary demonstrations, and we propose in a table a useful memorandum in daily practice. PMID- 17408137 TI - [Sjogren syndrome: a review of the literature and a case report]. AB - Sjogren syndrome is one of the most prevalent autoimmune diseases in which the body's immune system mistakenly attacks its own moisture producing glands. Although Sjogren syndrome occurs in all age groups in both women and men, women in their fourties are the most affected. Sjogren's syndrome can occur alone or in the presence of another connective tissue disease, respectively called primary and secundary Sjogren syndrome. When two of the three clinical hallmarks: keratoconjunctivitis sicca, xerostomia or connective tissue disease are present, Sjogren 's syndrome should be considered. To confirm the diagnosis of Sjogren's syndrome several tests are required. e.g. blood tests, ophthalmologic tests and oral tests. Rheumatologists have the primary responsibility for managing Sjogren's syndrome. Other specialists can treat the related symptoms. The incidence of lymphoma is higher in patients with Sjogren's syndrome than in the general population. Therefore patients must be monitored carefully for the development of related autoimmune diseases, lymphoma and other complications. Sjogren's syndrome is serious but generally not fatal if complications are diagnosed and treated early. PMID- 17408138 TI - [The current approach to hyper-sialorrhea]. AB - Drooling (sialorrhea or excessive salivation) is a common problem in neurologically impaired children and in adults who have Parkinson-disease. It is most commonly caused by poor oral and facial muscle control (aetiology) and worsened by some contributing factors. Treatment is best managed by a clinical team (speech pathologists, occupational therapists, dentists, otorhinolaryngologists) Treatment options range from conservative to more aggressive measures such as medication (anticholinergic medication, botulinum toxin type A injections in the glands), radiation and surgical therapy. PMID- 17408139 TI - [Saliva substitutes]. AB - Patients suffering from xerostomia lack oral comfort which reduces their ability to take up food properly and impairs their speech. Furthermore they suffer easily from caries and oral yeast infections. If it is impossible to stimulate a minimal salivary gland activity, salivary substitutes can be prescribed. Unfortunately, not all functions of saliva can be replaced by artificial products satisfactorily. Artificial saliva lacks a satisfactory efficacy with regards to lubrication, antimicrobial action and protection against demineralization. This can explain why these products are not widely known and their demand is very low. The optimal saliva substitute is not yet found. PMID- 17408140 TI - What counts? An ongoing question. PMID- 17408141 TI - Consensus offers real chance for reform. PMID- 17408142 TI - What does Canon Law say about the quality of sponsored works? PMID- 17408143 TI - The man and his work. Celebrating a life of prayer, study, and constant striving to be open to the spirit of truth. PMID- 17408144 TI - What is "prudential personalism"? Why does it matter? Fr. O'Rourke helped retrieve a vital ethical method in Catholic theology. PMID- 17408145 TI - Healing professions and Healing institutions. Frs. O'Rourke and Ashley have called for a "re-personalization" of health care. PMID- 17408146 TI - "Doing" ethics in an ecclesial context. What is health care ethics's connection to an understanding of the Catholic church? PMID- 17408147 TI - Fr. O'Rourke and beginning-of-life issues. His contributions are both foundational and practical. PMID- 17408148 TI - The ethics of end-of-life issues. Fr. O'Rourke has made wide-ranging contributions to thinking on the topic. PMID- 17408149 TI - Looking forward. A younger ethicist reflects on Fr. O'Rourke's contribution to Catholic health care ethics. PMID- 17408150 TI - Spiritual caregiving by health care professionals. Physicians, nurses, and other also have important contributions to make. PMID- 17408151 TI - Making access a priority. Ethics has a vital role in fostering collaboration in health care. PMID- 17408152 TI - What are the prospects for health care reform? PMID- 17408153 TI - Uniting to promote language access. PMID- 17408154 TI - Groundbreaking Delaware physicians: doctor Henry H. Stroud. PMID- 17408155 TI - So easy to forget. PMID- 17408156 TI - The Canadian Interventional Radiology Association. PMID- 17408157 TI - Conclusions and recommendations from the position paper on interventional radiology in Canada. PMID- 17408158 TI - Focal chronic pancreatitis mimicking pancreatic head carcinoma: are there suggestive features on ultrasound? AB - OBJECTIVE: Painless jaundice is one of the most common presentations of pancreatic head cancer. Chronic pancreatitis can also occasionally present with a mass or mass-like process in the pancreatic head, with the subsequent development of jaundice. In this retrospective review, we evaluate the clinical and imaging features of 22 patients presenting with painless jaundice, initially thought to have pancreatic head cancer and ultimately proven to have chronic focal pancreatitis, to determine whether there are any features on cross-sectional imaging to suggest the correct diagnosis. METHODS: Patients (n = 22) were identified from the medical and imaging records of more than 400 patients with an original diagnosis of pancreatic cancer who were seen at our institution from 1995 to 2003. Of the patients, 17 were men and 5 were women (age range 25 to 82 years, mean age 54 years). RESULTS: Initial ultrasound showed a large, hypoechoic, well-defined mass in the pancreatic head, varying in size from 3 to 7.5 cm; 14 of 22 masses were > 5 cm in maximal diameter. Diagnosis of focal pancreatitis was proven by surgical pathology in 14 cases and suggested by percutaneous biopsy in 3 cases; for all patients, prolonged imaging follow-up (at least 1 year) showed no disease progression or evidence of malignancy. The mean follow-up in our study group was 31 months (range 12 to 72 months). CONCLUSION: Male sex, large size of the pancreatic head mass (mean diameter 5.5 cm), and lack of atrophy of the pancreatic body and tail were the only features associated with chronic inflammation rather than neoplasm as an explanation for a pancreatic head mass. Chronic pancreatitis should be considered in the differential diagnosis of focal pancreatic masses, even in the absence of supporting clinical evidence. PMID- 17408159 TI - Computed tomography evaluation of traumatic rupture of the thoracic aorta: an outcome study. AB - OBJECTIVE: [corrected] To assess the long-term outcome of blunt trauma patients with suspected thoracic aortic or great vessel injury that was evaluated with contrast-enhanced chest computed tomography (CT). METHODS: We studied the outcome of 278 consecutive patients who received contrast-enhanced CT for blunt chest trauma with computerized searches of the regional trauma database, hospital medical records, universal government medical coverage plan billing records, and regional vital statistics databases. Data retrieved included patient demographics, mechanism of injury, status of the aorta and proximal great vessels at contrast-enhanced CT, hospital discharge diagnoses, and outpatient procedural billings with specific attention to aortic or great vessel injury. Median follow up was 615 days following the traumatic event. RESULTS: Six subjects demonstrated direct signs of aortic or proximal great vessel injury on contrast-enhanced chest CT, as follows: aortic pseudoaneurysm and intimal flap (n = 4), carotid artery dissection (n = 1), and aortic dissection (n = 1). All were surgically treated, except the patient with aortic dissection, who was treated medically. In the other subjects, contrast-enhanced CT was negative (n = 230) or showed isolated mediastinal hematoma (n = 42). The computerized searches of the medical databases showed that none of these 272 subjects had procedures for, or died from, aortic or great vessel injury during the follow-up period. CONCLUSION: Computerized searches of medical databases found no evidence of missed thoracic aortic or proximal great vessel injury in blunt trauma patients who were evaluated with contrast-enhanced chest CT. PMID- 17408160 TI - Recognizing and reporting vertebral fractures: reducing the risk of future osteoporotic fractures. AB - OBJECTIVE: Given the increasing evidence that vertebral fractures are underdiagnosed and not acted on, Osteoporosis Canada and the Canadian Association of Radiologists initiated a project to develop and publish a set of recommendations to promote and facilitate the diagnosis and reporting of vertebral fractures. OPTIONS: The identification of spinal fractures is not uniform. More than 65% of vertebral fractures cause no symptoms. It is also apparent that vertebral fractures are inadequately recognized when the opportunity for diagnosis arises fortuitously. It is to patients' benefit that radiologists report vertebral fractures evident on a chest or other radiograph, no matter how incidental to the immediate clinical indication for the examination. OUTCOMES: The present recommendations can help to close the gap in care in recognizing and treating vertebral fractures, to prevent future fractures and thus reduce the burden of osteoporosis-related morbidity and mortality, as well as fracture-related costs to the health care system. EVIDENCE: Several studies indicate that a gap exists in regard to the diagnosis of vertebral fractures and the clinical response following such diagnosis. All recommendations presented here are based on consensus. VALUES: These recommendations were developed by a multidisciplinary working group under the auspices of the Scientific Advisory Council of Osteoporosis Canada and the Canadian Association of Radiologists. BENEFITS, HARM, AND COSTS: Prevalent vertebral fractures have important clinical implications in terms of future fracture risk. Recognizing and reporting fractures incidental to radiologic examinations done for other reasons has the potential to reduce health care costs by initiating further steps in osteoporosis diagnosis and appropriate therapy. RECOMMENDATIONS: Physicians should be aware of the importance of vertebral fracture diagnosis in assessing future osteoporotic fracture risk. Vertebral fractures incidental to radiologic examinations done for other reasons should be identified and reported. Vertebral fractures should be assessed from lateral spinal or chest radiographs according to the semiquantitative method of Genant and colleagues. Grade II and Grade III fractures as classified by this method should be given the greatest emphasis. Semiquantitative fracture recognition should include the recognition of changes such as loss of vertebral end-plate parallelism, cortical interruptions, and quantitative changes in the anterior, midbody, and posterior heights of vertebral bodies. When spine radiographs are performed to assess the presence of vertebral fractures, anteroposterior examinations may assist in the initial evaluation. The standard follow-up need only consist of single lateral views of the thoracic and lumbar spine that include T4 to L4 vertebrae. The radiographic technique described in this paper, or a technique of comparable efficacy, should be used. Dual X-ray absorptiometry examinations that include lateral spinal morphological assessments (vertebral fracture assessment) may contribute to fracture recognition. Educational material about the clinical importance of vertebral fracture recognition as a potential indicator of future osteoporotic fracture risk with its associated morbidity and mortality should be directed to all physicians. VALIDATION: Recommendations were based on consensus opinion. PMID- 17408161 TI - Age-predicted values for lumbar spine, proximal femur, and whole-body bone mineral density: results from a population of normal children aged 3 to 18 years. AB - We measured areal bone mineral density (BMD) with dual-energy X-ray absorptiometry (DXA) at the lumbar spine and the proximal femur and for the total body in 179 subjects (91 girls and 88 boys) with no known disorders that might affect calcium metabolism. Results are also reported for lumbar spine bone mineral content (BMC) and for the derived variable, bone mineral apparent density (BMAD). Expected-for-age values for each variable were derived for boys and girls by using an expression that represented the sum of a steady increase due to growth plus a rapid increase associated with puberty. Normal ranges were derived by assuming that at least 95% of children would be included within 1.96 population standard deviations (SD) of the expected-for-age value. The normal range for lumbar spine BMD derived from our population of children was compared with previously published normal ranges based on results obtained from different bone densitometers in diverse geographic locations. The extent of agreement between the various normal ranges indicates that the derived expressions can be used for reporting routine spine, femur, and whole-body BMD measurements in children and adolescents. The greatest difference in expected-for-age values among the various studies was that arising from intermanufacturer variability. The application of published conversion factors derived from DXA measurements in adults did not account fully for these differences, especially in younger children. PMID- 17408162 TI - Whole-body bone mineral content, lean body mass, and fat mass measured by dual energy X-ray absorptiometry in a population of normal Canadian children and adolescents. AB - Measurements of body composition have evident value in evaluating growing children and adolescents, and dual-energy X-ray absorptiometry (DXA) is a tool that provides accurate measurements of whole-body bone mineral content (WBBMC), lean body mass (LBM), and fat mass (FM). To interpret such measurements in the context of ill health, normative values must be available. Such information could be expected to be regionally specific because of differences in ethnic, dietary, and physical activity determinants. In this study, DXA was performed with Hologic densitometers in normal girls (n = 91) and boys (n = 88) between 3 and 18 years of age. The derivation of normal ranges is presented for boys and girls. The correlation of the sum of WBBMC, LBM, and FM with directly measured body weight was almost perfect (r > 0.997). As expected, FM and body mass index correlated strongly. The normal values for WBBMC, LBM, and FM from this study are compared with other Canadian data and with published normative data from Argentina and the Netherlands, all of which use different densitometers. The results of this study allow the calculation of z scores for each facet of body composition and facilitate the use of DXA to report routine evaluations of body composition in children and adolescents. PMID- 17408163 TI - Answer to case of the month #115. Sclerosing encapsulating peritonitis with calcification. PMID- 17408164 TI - Answer to case of the month #116. Aortocaval fistula. PMID- 17408165 TI - [Lithiasis today]. PMID- 17408166 TI - [Research study on renal targeted chemotherapy with magnetic harpoons and intravenous administration of ferro-carbon nanoparticles]. AB - OBJECTIVES: The use of nanoparticules for drug transport is one of the topics with priority interest within the field of biomedical research. Our objective is to show the initial results of an innovative method to focalize drug carrier ferro-carbon nanoparticules to solid organs. We obtained and characterized various types of ferrous magnetic nanoparticules and studied their behaviour in vitro and in vivo in laboratory animals with intrarenal magnetic targets laparoscopically implanted. METHODS: Using a plasma arch we obtained ferro-carbon nanoparticules with the ability to absorb and deliver doxorubicin, showing an excellent behaviour in in vitro rheological studies. Under general anesthesia and control we inserted a gold covered magnetic microharpoon in the left kidney of New Zealand rabbits. At the same time we injected intravenously different doses of various types of nanoparticules. The animals were sacrified ofter pre established times and pathologic studies of their kidneys, spleens, livers, lungs and bone marrow were carried out. RESULTS: After selection of the most adequafe nanoparticules for our purposes, we ascertained significant differences in the distribution of nanoparticules in postmortem studies, with accumulation in the magnetic target and surrounding renal parenchyma. Nevertheless, the reticuloendothelial system retains a great amount of the injected dose. CONCLUSIONS: Although our magnetic focalization system is effective, nanoparticule temporary protection systems should be tested to allow us avoid the action of the immune system. PMID- 17408167 TI - [Diagnosis and management of patients with overactive bladder syndrome in urology clinics and general practitioner clinics in Spain]. AB - OBJECTIVES: Despite its high prevalence and its important impact on patient's life, overactive bladder is a disorder poorly known and not usually tackled in daily clinical practice. The aim of this study is to know the main procedures and techniques used by both urologists and general practitioners to diagnose overactive bladder in usual clinical practice in Spain. METHODS: A standardized questionnaire was administered to 748 physicians specialized in urology and to 696 primary core physicians or general practitioners. The questionnaire collected information about the management of patients who attended surgery with urinary symptoms suggesting overactive bladder. RESULTS: A total of 64.8% of urologists argued they followed a standardized protocol in order to diagnose patients suffering from overactive bladder. Concerning primary care physicians, 58.4% of them admitted not referring patients with urinary symptoms suggesting overactive bladder to specialist. Up to 77.8% of urologists confirmed that referred patients from primary care had no previous exam or diagnosis. Health history, physical examination and urinalysis were the usual diagnosis procedures in clinical practice conditions for both specialities. Other instruments such as the micturition diary or the questionnaires to assess symptoms or patients' quality of life are still little used for the diagnosis and management of overactive bladder. CONCLUSIONS: Although overactive bladder is a condition that causes a high impairment in the quality of life (QoL) and daily activities of those patients suffering from it, it still remains poorly tackled by doctors. Therefore, it is important to define strategies to detect its symptoms in the clinical practice. PMID- 17408168 TI - [Results of the surgical treatment of benign prostatic hyperplasia in geriatric patients]. AB - OBJECTIVES: To evaluate the effect of age in morbidity-mortality and outcomes of the surgical treatment of benign prostatic hyperplasia (BPH). METHOD: We performed a retrospective analysis of the medical records of 305 patients over 70 years undergoing surgery for BPH between 1999-2003, grouped in four categories depending on ages 70-80 years and over 80, and type of surgery (transurethral resection or open prostatectomy). The variables evaluated were: American society of anesthesiologists (ASA) operative risk classification, surgical indication, preoperative factors, postoperative outcome and follow-up. Statistical analysis was carried out with the Anova and chi-square tests (p = 0.05). RESULTS: Open prostatectomy was performed in 59.1% of the cases and transurethral resection in the other 40.9%. 98.4% of the patients showed some degree of comorbidity. Indwelling transurethral catheter was the main clinical feature of patients undergoing surgery over 80 years of age; the main characteristic in patients between 70-80 years was the existence of severe symptoms. Urological complications appeared more frequently than general complications (14% vs. 10.1%). The most frequent urological complications were mild, mainly inflammatory infectious diseases. Urological and general complications were more frequently observed in the open prostatectomy group, with no differences between ages. Patient satisfaction was high in the four groups (84.8%-96.2%). No case of perioperative mortality happened and only one patient died in the immediate post operative period. CONCLUSIONS: We think the differences observed cannot be attributed to chronological age as clinical risk factor. Therefore, these patients could benefit of a definitive surgical treatment which will improve their quality of life. PMID- 17408169 TI - [Intravesical therapy: Influence on superficial bladder cancer progression]. AB - OBJECTIVES: To review the influence of various intravesical treatments on superficial bladder cancer progression. METHODS: We retrospectively reviewed 473 superficial bladder neoplasias. Based on diagnosis and transurethral resection of bladder tumor (TUR BT), and depending on pathology, we proposed different treatment and follow-up schemes, with BCG in 139 cases and intravesical chemotherapy, mainly Mytomicin C, in 80. RESULTS: Overall, the use or not of bladder instillations did not show statistically significant differences in time to progression. The use of intravesical therapy and maintenance therapy was an independent risk factor for disease-free time to progression when compared with induction. CONCLUSIONS: The use of maintenance intravesical therapy with BCG resulted in a delayed progression of superficial bladder cancer, so that it seems the most effective treatment, mainly in moderate-high risk tumors. PMID- 17408170 TI - [Intravesical therapy: Influence on superficial bladder cancer recurrence]. AB - OBJECTIVES: To review the influence of various intravesical treatments on superficial bladder cancer recurrence. METHODS: We retrospectively reviewed 473 superficial bladder neoplasias. Based on diagnosis and transurethral resection of bladder tumor (TUR BT), and depending on pathology, we proposed different adjuvant treatment and follow-up schemes. RESULTS: The following factors were independent risk factors for a longer recurrence-free time: intravesical instillations, high dose BCG, and maintenance therapy in comparison to induction. CONCLUSIONS: The use of intravesical instillations independently increases the time to recurrence. It was demonstrated that recurrence was significantly later when high dose BCG and maintenance therapy were employed. PMID- 17408171 TI - [New technique for the repair of anterior pelvic floor compartment defects using a synthetic implant with biological coverage: approach, fixation and transobturator anchoring]. AB - Transobturator systems for anterior vaginal wall prolapse repair exemplify the current trend in pelvic floor surgery. They may be considered an approach and also a mesh fixation system, in opposition to free mesh cystocele repair where they work by the creation of fibrotic tissue after mesh implant (biological or synthetic). OBJECTIVES: To describe the elements of the Avaulta anterior system, its indications and the surgical technique to implant it and adequately adjust it. METHODS: The operation has five steps: (1) midline vaginal incision and mucosal dissection, (2) Obturator foramen identification, design and performance of 2 superior mini incisions in both the genitofemoral folds, and another two 3 cm below and 1-2 cm lateral to them, (3) Needle introduction and passage through the upper portion of the obturator foramen, parallel to the ischiopubic ramus, and once past needle charging with the arm of the implant, (4) Needle introduction and passage from the inferior incisions vertically through the inferior portion of the obturator foramen, directing the needle with bimanual control to the theoretical localization of the uterine.cervix, with connection and charging of the inferior arm of the mesh. (5) Tension free adjustment of the mesh and closure of the incisions. CONCLUSIONS: (1) It is a reproducible technique that adequately corrects the anterior vaginal compartment defects. (2) The design and technology of Avaulta aims to correct the anterior compartment defects, based on the principles of ideal mesh. PMID- 17408172 TI - [Importance of the neurophysiological evoked potential register during the implantation of a sacral neuromodulator]. AB - OBJECTIVES: Sacral neuromodulation is a new treatment option in patients with urge incontinence, urgency-frequency and urinary retention refractory to the pharmacological therapy. The electrophysiological monitoring provides direct and immediate feedback about the function of nerves. We evaluated the utility of the electrophysiological monitoring to assert the correct position of the lead at the third sacral root (S3). METHOD: We registered the sacral-vesicourethral and sacral plantar evoked potentials in two patients (a 55 years old man and a 37 year-old woman) during the implant of a sacral neuromodulator (Interstim, Medtronic, MN USA). RESULTS: We registered vesico-urethral and sacroplantar potentials in response to sacral stimulation. We found vesical evoked potentials after the stimulation of S3 and S4 and plantar responses after the stimulation of S2 and S3. The vesico-urethral evoked potentials had a latency of 25.10 mseg in our first case and 28.52 mseg in the second. The sacroplantar evoked potentials had a latency of 20.80 mseg and 22.46 mseg respectively. We observed the evoked potential register during the surgery prior to the visual motor and sensory responses with a lower intensity of stimulation (average 3.5 mAmp). The only sacral root in which we could find both responses was in S3. CONCLUSION: The electrophysiological monitoring was helpful in identifying the right position of the lead at the correct sacral root. It avoided the necessity of high stimulation intensity during the procedure. PMID- 17408173 TI - [Azoospermia should not be given as the result of vasectomy]. AB - OBJECTIVES: Vasectomy is a surgical method of male contraception. Azoospermia is offered as result of the technique and this is not always attained, resulting in legal matters. The purpose of this study is to know the number of semen samples needed to discharge a patient after intervention. To identify sperm count on semen analysis at time of discharge. METHODS: Retrospective study of men who underwent vasectomy in a 15-month period with a 2 year follow up. Consecutive semen analyses up to 5 samples were measured at 2 to 3 months interval in all men who had persistence of spermatozoa. RESULTS: 618 men were intervened, 106 did not bring semen to the laboratory (17%), 2 (0.39%) presented motile sperm and were considered a failure of the technique and excluded. 510 men completed controls. 316 (61.9%) were azoospermic in the first sperm analysis, 74 (14.5%) in the second, 27 (5.2%) in the third, 6 (1.2%) in the fourth and one (0,.%) in the fifth analysis. The remaining 86 men (16.8%) had persistence of immotile sperm in the ejaculate and were less than 100,000/ml. No pregnancy was reported during 2 years follow up or after. CONCLUSIONS: Five or more semen analysis can be made after the surgery. Persistence of immotile sperm in the ejaculate is frequent and may exist for a long period afterwards. Immotile sperm count of 100,000/ml or less should be accepted as result of the procedure. The patient should be informed about the fact that persistent immotile sperm can be found in his semen. In the informed consent azoospermia should not be a concern as it is frequent to find immotile sperm in the ejaculate and this is an acceptable issue. As with other contraceptive methods, vasectomy should be offered as a safe method although clearly stating that the possibilities of failure do exist. PMID- 17408174 TI - [Retroperitoneal laparoscopic lymphadenectomy for stage I non seminomatous testicular cancer]. AB - OBJECTIVES: We report a series of patients with stage I non seminomatous testicular cancer in whom laparoscopic retroperitoneal lymphadenectomy was carried out. METHODS: Between March 1993 and January 2004 100 and patients with the diagnosis of stage I non seminomatous testicular cancer underwent laparoscopic retroperitoneal lymphadenectomy (LRPL). All operations were performed by the same surgeon (O.A.C). Clinical data were collected prospectively in a database. Patients' demographic data, operative time, estimated blood loss, hospital stay, operative and post operative complications, convention rate and follow-up were all analyzed. RESULTS: 111 LRPL were performed in 111 patients with a mean age of 29.3 years (range 15-44 years). 61 procedures were right-LRPL and 50 left-LRPL. Mean operative time was 138 minutes (range 60-300 minutes). Introoperative complications appeared in 10 cases (9%), 9 vascular complications (8.1%) and 1 duodenal lesion (0.9%). Conversion to open surgery was necessary in three patients (2.7%). Mean hospital stay was 43.3 hours (range 24-120). 5 patients (4.5%) had tumor recurrence, with a mean follow-up of 30 months (range 24-94 months). Recurrences were localized in the retroperitoneum, mediastinum and lungs. Recurrence time varied between four and 64 months. CONCLUSIONS: LRPL offers oncological results equivalent to open surgery, with the advantage of being minimally invasive and its consequent decrease in morbidity. The laparoscopic retroperitoneal lymphadenectomy is a good option for the management of patients with stage 1 testicular cancer. PMID- 17408175 TI - [Bechet's disease. Case report]. AB - OBJECTIVE: To report one case of a rare disease. METHODS: 39-year-old male patient presenting with the suspicion of sexually transmitted disease. We show several pictures of a clinical case diagnosed and treated in our hospital. We performed a bibliographic review on the disease and present its etiology, diagnosis and treatment. RESULTS: We concluded it was Bechet's disease because of the past medical history, clinical presentation, and diagnostic tests (positive pathergy test). CONCLUSIONS: Although it is a rare disease, we should always think of the diagnosis of Bechet's disease in a patient with genital ulcers non responsive to treatment. PMID- 17408176 TI - [Goldenhar syndrome associated to posterior urethral valves]. AB - OBJECTIVE: To report a case with Goldenhar syndrome with posterior urethral valves. METHODS: Goldenhar syndrome (oculoauriculovertebral dysplasia) is a condition featuring the following triad of anomalies: ocular abnormalities (epibulbar dermoids, coloboma), otic anomalies (low set ears, auricular appendage) and/or vertebral anomalies. Multiple malformations, including congenital heart, brain and renal disease. RESULTS: Posterior urethral valves should be treated with primary valve ablation. CONCLUSIONS: It is necessary to perform a careful evaluations of general malformations, especially renal malformations. PMID- 17408177 TI - [Adult's mesoblastic nephroma. Report of a case with aggressive course]. AB - OBJECTIVE: Mesoblastic nephroma is a rare renal neoplasia mainly diagnosed in the first three months of life; there is an adult type with pathologic similarities but it has its own features. METHODS: We report the case of a 71-year-old female patient with the diagnosis of adult mesoblastic nephroma, the clinical outcome of which was ominous. RESULTS: The presence of epithelial elements with tubular conformation surrounded by a spindle cell component is greatly useful to perform the differential diagnosis between this entity and others of greater clinical significance. CONCLUSIONS: Although scarce, there are cases in the bibliography similar to ours, with an aggressive behaviour that maybe require a more aggressive treatment, and not the conservative one traditionally used for these tumors. PMID- 17408178 TI - [MRI for the diagnosis of Leydig cell testicular tumors]. AB - OBJECTIVE: To determine the diagnostic usefulness of MRI in the diagnosis of Leydig cell testicular tumor. METHODS/RESULTS: Male patient consulting for infertility. Testicular ultrasound and MRI were performed. Testicular MRI showed a hypointense lesion in T2 powered sequences with intense, homogeneous enhance after contrast administration. CONCLUSIONS: MRI is considered an excellent diagnostic test for the management of patients with testicular diseases of difficult diagnosis on ultrasound, contributing to define the lesion, a Leydig cell testicular tumor in this case. PMID- 17408179 TI - [Follicular cystitis. Case report and bibliographic review]. AB - OBJECTIVE: To report one case of chronic follicular cystitis and to perform a bibliographic review on this pathology. CASE REPORT: We report a case of a 70 year-old woman with an irritative voiding syndrome over an eight-month period, with several previous episodes of urinary tract infection treated by her family doctor. Bullous lesions were found in the bladder mucosa on cystoscopy. Histological tests showed lymphoid follicles at the level of the bladder mucosa, leading to the diagnosis of follicular cystitis. RESULTS: Medical treatment was carried out with ciprofloxacin, vitamin A, and prednisone, which led to symptom remission. CONCLUSIONS: Follicular cystitis is an entity that belongs to the group of chronic cystopathies; it is a non-specific chronic inflammatory disease characterized by the presence of large number of plasmatic cells and lymphocytes in lymphoid follicles within the bladder mucosa and submucosa; pathologic study is necessary for the final diagnosis of this entity. PMID- 17408180 TI - [Metachronic testicular tumor secondary to invasive bladder carcinoma]. AB - OBJECTIVE: To report a rare case of testicular metastasis secondary to an infiltrative transitional cell carcinoma years after radical surgery. METHODS: CASE REPORT: 71-year-old male patient with history of infiltrative bladder tumor, status post radical cystoprostatectomy with urethrectomy. Seven years after surgery he presents with pain and swelling in the right testicle. Inguinal orchyectomy was carried out with the pathologic diagnosis of testicular metastasis of a high-grade transitional cell carcinoma. We perform a bibliographic review. RESULTS: The patient is disease-free twelve months after orchyectomy. CONCLUSIONS: Apart from leukemia and lymphoma, testicular metastases are extremely rare. The existence of testicular metastases means a metastatic dissemination to other organs, therefore chemotherapy could improve prognosis. PMID- 17408181 TI - [Acute renal failure secondary to the transurethral resection of the prostate syndrome]. AB - OBJECTIVE: Complications of transurethral resection of the prostate (TURP) related to hypotonicity and hypervolemia were well described in the literature. However acute renal failure, known to be a clinical presentation of some TURP syndromes was less discussed. METHODS: We report a case of oliguric acute renal failure as a major complication after TURP, with a discussion of a possible mechanism for the disorder. RESULTS: The mechanism by which renal failure developed in our patient is not entirely clear, but most likely is due to hemolysis. Other factors such as hemodynamic alterations, hypotension and rabdomyolisis are also suspected based on analysed data. CONCLUSION: We believe that a detailed evaluation before and after TURP will allow to reduce the incidence of this abnormality and to prevent its occurrence. PMID- 17408182 TI - [Anterior urethral rupture without involvement of cavernous bodies]. PMID- 17408183 TI - [Renal artery aneurysm]. PMID- 17408184 TI - A rare complication associated to a suprapubic cystostomy: catheter knotting. AB - We report a rare case of spontaneous catheter knotting in the setting of a suprapubic cystostomy and also review previous reports and the rationale behind this event. PMID- 17408185 TI - Secondary testicular plasmocytoma. AB - OBJECTIVE: [corrected] To report a rare case of secondary testicular plasmocytoma in the context of a testicular mass. METHOD: We introduced a patient with plasmocytoma and a testicular infiltration of his systemic affection. RESULT: An inguinal orquiectomy was performed to treat the testicular infiltration and no recurrence of the process was seen after 6 months of follow up. CONCLUSIONS: Plasmacytoma is a plasma cell tumor that involves bone marrow or extramedullary sites. Testicular infiltration by plasma cells such as plasmacytoma happens in only 2% of cases of all plasma cell neoplasm PMID- 17408186 TI - Methylmercury exposure and health effects in humans: a worldwide concern. AB - The paper builds on existing literature, highlighting current understanding and identifying unresolved issues about MeHg exposure, health effects, and risk assessment, and concludes with a consensus statement. Methylmercury is a potent toxin, bioaccumulated and concentrated through the aquatic food chain, placing at risk people, throughout the globe and across the socioeconomic spectrum, who consume predatory fish or for whom fish is a dietary mainstay. Methylmercury developmental neurotoxicity has constituted the basis for risk assessments and public health policies. Despite gaps in our knowledge on new bioindicators of exposure, factors that influence MeHg uptake and toxicity, toxicokinetics, neurologic and cardiovascular effects in adult populations, and the nutritional benefits and risks from the large number of marine and freshwater fish and fish eating species, the panel concluded that to preserve human health, all efforts need to be made to reduce and eliminate sources of exposure. PMID- 17408187 TI - Effects of environmental methylmercury on the health of wild birds, mammals, and fish. AB - Wild piscivorous fish, mammals, and birds may be at risk for elevated dietary methylmercury intake and toxicity. In controlled feeding studies, the consumption of diets that contained Hg (as methylmercury) at environmentally realistic concentrations resulted in a range of toxic effects in fish, birds, and mammals, including behavioral, neurochemical, hormonal, and reproductive changes. Limited field-based studies, especially with certain wild piscivorous bird species, e.g., the common loon, corroborated laboratory-based results, demonstrating significant relations between methylmercury exposure and various indicators of methylmercury toxicity, including reproductive impairment. Potential population effects in fish and wildlife resulting from dietary methylmercury exposure are expected to vary as a function of species life history, as well as regional differences in fish-Hg concentrations, which, in turn, are influenced by differences in Hg deposition and environmental methylation rates. However, population modeling suggests that reductions in Hg emissions could have substantial benefits for some common loon populations that are currently experiencing elevated methylmercury exposure. Predicted benefits would be mediated primarily through improved hatching success and development of hatchlings to maturity as Hg concentrations in prey fish decline. Other piscivorous species may also benefit from decreased Hg exposure but have not been as extensively studied as the common loon. PMID- 17408188 TI - A synthesis of progress and uncertainties in attributing the sources of mercury in deposition. AB - A panel of international experts was convened in Madison, Wisconsin, in 2005, as part of the 8th International Conference on Mercury as a Global Pollutant. Our charge was to address the state of science pertinent to source attribution, specifically our key question was: "For a given location, can we ascertain with confidence the relative contributions of local, regional, and global sources, and of natural versus anthropogenic emissions to mercury deposition?" The panel synthesized new research pertinent to this question published over the past decade, with emphasis on four major research topics: long-term anthropogenic change, current emission and deposition trends, chemical transformations and cycling, and modeling and uncertainty. Within each topic, the panel drew a series of conclusions, which are presented in this paper. These conclusions led us to concur that the answer to our question is a "qualified yes," with the qualification being dependent upon the level of uncertainty one is willing to accept. We agreed that the uncertainty is strongly dependent upon scale and that our question as stated is answerable with greater confidence both very near and very far from major point sources, assuming that the "global pool" is a recognizable "source." Many regions of interest from an ecosystem-exposure standpoint lie in between, where source attribution carries the greatest degree of uncertainty. PMID- 17408189 TI - Recovery of mercury-contaminated fisheries. AB - In this paper, we synthesize available information on the links between changes in ecosystem loading of inorganic mercury (Hg) and levels of methylmercury (MeHg) in fish. Although it is widely hypothesized that increased Hg load to aquatic ecosystems leads to increases in MeHg in fish, there is limited quantitative data to test this hypothesis. Here we examine the available evidence from a range of sources: studies of ecosystems contaminated by industrial discharges, observations of fish MeHg responses to changes in atmospheric load, studies over space and environmental gradients, and experimental manipulations. A summary of the current understanding of the main processes involved in the transport and transformation from Hg load to MeHg in fish is provided. The role of Hg loading is discussed in context with other factors affecting Hg cycling and bioaccumulation in relation to timing and magnitude of response in fish MeHg. The main conclusion drawn is that changes in Hg loading (increase or decrease) will yield a response in fish MeHg but that the timing and magnitude of the response will vary depending of ecosystem-specific variables and the form of the Hg loaded. PMID- 17408190 TI - Socioeconomic consequences of mercury use and pollution. AB - In the past, human activities often resulted in mercury releases to the biosphere with little consideration of undesirable consequences for the health of humans and wildlife. This paper outlines the pathways through which humans and wildlife are exposed to mercury. Fish consumption is the major route of exposure to methylmercury. Humans can also receive toxic doses of mercury through inhalation of elevated concentrations of gaseous elemental mercury. We propose that any effective strategy for reducing mercury exposures requires an examination of the complete life cycle of mercury. This paper examines the life cycle of mercury from a global perspective and then identifies several approaches to measuring the benefits of reducing mercury exposure, policy options for reducing Hg emissions, possible exposure reduction mechanisms, and issues associated with mercury risk assessment and communication for different populations. PMID- 17408191 TI - The Madison declaration on mercury pollution. AB - This declaration summarizes the scientific and technical conclusions presented by four expert panels in their critical synthesis manuscripts and in plenary sessions at the Eighth International Conference on Mercury as a Global Pollutant, convened in Madison, Wisconsin, USA, on 6-11 August 2006. The 1150 registered participants in this conference constituted a diverse, multinational body of scientific and technical expertise on environmental mercury pollution. This declaration conveys the panels' principal findings and their consensus conclusions on key policy-relevant questions concerning atmospheric sources of mercury, methylmercury exposure and its effects on humans and wildlife, socioeconomic consequences of mercury pollution, and recovery of mercury contaminated fisheries. PMID- 17408192 TI - Epidemiological transitions and the changing face of medical geology. PMID- 17408193 TI - Iodine deficiency: an ancient problem in a modern world. PMID- 17408194 TI - Natural minerogenic dust and human health. PMID- 17408195 TI - Global impacts of geogenic arsenic: a medical geology research case. PMID- 17408196 TI - Blackfoot disease in Taiwan: its link with inorganic arsenic exposure from drinking water. PMID- 17408197 TI - Radon: sources, health risks, and hazard mapping. PMID- 17408198 TI - Selenium deficiency and endemic heart failure in China: a case study of biogeochemistry for human health. PMID- 17408199 TI - Selenium geochemistry and health. PMID- 17408200 TI - Health effects of toxic organic substances from coal: toward "panendemic" nephropathy. PMID- 17408201 TI - Health impacts of coal: facts and fallacies. PMID- 17408202 TI - Metal biology: aspects of beneficial effects. PMID- 17408203 TI - Mobilization of mercury and methylmercury from forest soils after a severe storm fell event. PMID- 17408204 TI - Medical geology: an opportunity for the future. PMID- 17408205 TI - PCPs need to step up their care of patients with CKD. PMID- 17408206 TI - Research underscores the value of routine HIV screening, but barriers remain. AB - HIV screening offers good value, but obstacles remain, That's the message of at least one new study that provides a solid foundation for new guidelines, recommending that HIV screening be integrated into routine care. However, huge obstacles--including lack of funding and legislative barriers--continue to stand in the way of the kind of broad screening practices that policy experts agree are needed. PMID- 17408207 TI - Clinic uses chronic care model to tackle diabetes. AB - The chronic care model establishes a path to improvement--even at the practice level. It's a highly esteemed model for chronic care management, but even large organizations struggle to implement all of the elements that comprise the chronic care model. However, it can be done at the practice level. See how one Seattle based clinic used it to produce an array of improvements in diabetes care. PMID- 17408208 TI - Unique approach offers guidance to patients with low-prevalence/ high-cost conditions. AB - A unique intervention delivers behavioral health support to patients with less prevalent, but high-cost conditions. Not nearly as many patients suffer from Crohn's disease or rheumatoid arthritis as diabetes, but Newark, NJ-based Horizon Blue Cross and Blue Shield believes it can nonetheless improve care for these types of patients-and recoup an ROI--by providing them with an intervention that emphasizes psychosocial support. PMID- 17408209 TI - Liposomes as an ocular delivery system for acetazolamide: in vitro and in vivo studies. AB - The purpose of this study was to formulate topically effective controlled release ophthalmic acetazolamide liposomal formulations. Reverse-phase evaporation and lipid film hydration methods were used for the preparation of reverse-phase evaporation (REVs) and multilamellar (MLVs) acetazolamide liposomes consisting of egg phosphatidylcholine (PC) and cholesterol (CH) in the molar ratios of (7:2), (7:4), (7:6), and (7:7) with or without stearylamine (SA) or dicetyl phosphate (DP) as positive and negative charge inducers, respectively. The prepared liposomes were evaluated for their entrapment efficiency and in vitro release. Multilamellar liposomes entrapped greater amounts of drug than REVs liposomes. Drug loading was increased by increasing CH content as well as by inclusion of SA. Drug release rate showed an order of negatively charged > neutral > positively charged liposomes, which is the reverse of the data of drug loading efficiency. Physical stability study indicated that approximately 89%, 77%, and 69% of acetazolamide was retained in positive, negative, and neutral MLVs liposomal formulations up to a period of 3 months at 4 degrees C. The intraocular pressure (IOP)-lowering activity of selected acetazolamide liposomal formulations was determined and compared with that of plain liposomes and acetazolamide solution. Multilamellar acetazolamide liposomes revealed more prolonged effect than REVs liposomes. The positively charged and neutral liposomes exhibited greater lowering in IOP and a more prolonged effect than the negatively charged ones. The positive multilamellar liposomes composed of PC:CH:SA (7:4:1) molar ratio showed the maximal response, which reached a value of -7.8 +/- 1.04 mmHg after 3 hours of topical administration. PMID- 17408210 TI - Elucidation of spheroid formation with and without the extrusion step. AB - Spheroid formation mechanisms were investigated using extrusion-spheronization (ES) and rotary processing (RP). Using ES (cross-hatch), ES (teardrop), and RP (teardrop), spheroids with similar mass median diameter (MMD) and span were produced using equivalent formulation and spheronization conditions. During spheronization, the teardrop-studded rotating frictional surface, with increased peripheral tip speed and duration, produced spheroids of equivalent MMD and span to those produced by the cross-hatch rotating frictional plate surface. The roundness of these spheroids was also similar. RP required less water to produce spheroids of MMD similar to that of spheroids produced by ES. However, these RP spheroids were less spherical. Image analysis of 625 spheroids per batch indicated that the size distribution of RP spheroids had significantly greater SD, positive skewness, and kurtosis. Morphological examination of time-sampled spheroids produced by ES indicated that spheroid formation occurred predominantly by attrition and layering, while RP spheroids were formed by nucleation, agglomeration, layering, and coalescence. RP produced spheroids with higher crushing strength than that of ES-produced spheroids. The amount of moisture lost during spheronization for spheroids produced by ES had minimal influence on their eventual size. Differences in process and formulation parameters, in addition to size distribution and observed morphological changes, enabled a greater understanding of spheroid formation and methods to optimize spheroid production. PMID- 17408211 TI - Gastroretentive drug delivery system of carbamazepine: formulation optimization using simplex lattice design: a technical note. PMID- 17408212 TI - Eudragit-coated pectin microspheres of 5-fluorouracil for colon targeting. AB - An objective of the present investigation was to prepare and evaluate Eudragit coated pectin microspheres for colon targeting of 5-fluorouracil (FU). Pectin microspheres were prepared by emulsion dehydration method using different ratios of FU and pectin (1:3 to 1:6), stirring speeds (500-2000 rpm) and emulsifier concentrations (0.75%-1.5% wt/vol). The yield of preparation and the encapsulation efficiencies were high for all pectin microspheres. Microspheres prepared by using drug:polymer ratio 1:4, stirring speed 1000 rpm, and 1.25% wt/vol concentration of emulsifying agent were selected as an optimized formulation. Eudragit-coating of pectin microspheres was performed by oil-in-oil solvent evaporation method using coat:core ratio (5:1). Pectin microspheres and Eudragit-coated pectin microspheres were evaluated for surface morphology, particle size and size distribution, swellability, percentage drug entrapment, and in vitro drug release in simulated gastrointestinal fluids (SGF). The in vitro drug release study of optimized formulation was also performed in simulated colonic fluid in the presence of 2% rat cecal content. Organ distribution study in albino rats was performed to establish the targeting potential of optimized formulation in the colon. The release profile of FU from Eudragit-coated pectin microspheres was pH dependent. In acidic medium, the release rate was much slower; however, the drug was released quickly at pH 7.4. It is concluded from the present investigation that Eudragit-coated pectin microspheres are promising controlled release carriers for colon-targeted delivery of FU. PMID- 17408213 TI - Improving the tablet characteristics and dissolution profile of ibuprofen by using a novel coprocessed superdisintegrant: a technical note. PMID- 17408214 TI - Formulation and evaluation of ketorolac tromethamine-loaded albumin microspheres for potential intramuscular administration. AB - The objective of this work was to prepare and evaluate ketorolac tromethamine loaded albumin microspheres using a factorial design. Albumin microspheres were prepared by emulsion cross-linking method. Selected formulations were characterized for their entrapment efficiency, particle size, surface morphology, and release behavior. Analysis of variance (ANOVA) for entrapment efficiency indicated that entrapment efficiency is best fitted to a response surface linear model. From the statistical analysis it was observed that as the drug:polymer (D:P) ratio and volume of glutaraldehyde increased, there was a significant increase in the encapsulation efficiency. Scanning electron microscopy of the microspheres revealed a spherical, nonporous and uniform appearance, with a smooth surface. Based on the entrapment efficiency and physical appearance, 9 formulations were selected for release study. The maximum particle size observed was below 40 microm. The release pattern was biphasic, characterized by an initial burst effect followed by a slow release. All selected microspheres, except those having less polymer proportion (D:P ratio is 1:1), exhibited a prolonged release for almost 24 hours. On comparing r (2) values for Higuchi and Peppas kinetic models, different batches of microspheres showed Fickian, non Fickian, and diffusion kinetics. The release mechanism was regulated by D:P ratio and amount of cross-linking agent. From the experimental data obtained with respect to particle size and extent of drug release, it could be concluded that the prepared microspheres are useful for once-a-day intramuscular administration of ketorolac tromethamine. PMID- 17408215 TI - Development of a system for simultaneous dissolution studies and magnetic resonance imaging of water transport in hydrodynamically balanced systems: a technical note. PMID- 17408216 TI - Degradation kinetics and mechanism of RH1, a new anti-tumor agent: a technical note. PMID- 17408217 TI - Suppression of agglomeration of ciprofloxacin-loaded human serum albumin nanoparticles. AB - The present study is aimed at developing and exploring the use of pectin in suppression of agglomeration of ciprofloxacin-loaded human serum albumin (HSA) nanoparticles. The HSA-pectin nanoparticles loaded with ciprofloxacin were prepared by the pH-coacervation method, and various physicochemical parameters such as particle size, morphology, zeta-potential, electrolyte-induced flocculation, pH-dependent zeta-potential, drug loading, in vitro drug release, and stability of nanoparticles, were evaluated. The size of the HSA-pectin nanoparticles (F3) was found to be 180 to 290 nm. The HSA nanoparticles were modified with pectin when the critical flocculation concentration of nanoparticles in Na(2)SO(4) solution was increased from 0.3 M to 0.9 M. The isoelectric points of the formed nanoparticles were found to be relatively lower between pH values 3 and 6. Pectin may be used as a pharmaceutical additive for the suppression of particle agglomeration in HSA nanoparticles, and the effect may be attributed to the pectin segments present on the surface of nanoparticles. PMID- 17408218 TI - Transdermal therapeutic system of carvedilol: effect of hydrophilic and hydrophobic matrix on in vitro and in vivo characteristics. AB - The purpose of this research was to develop a matrix-type transdermal therapeutic system containing carvedilol with different ratios of hydrophilic and hydrophobic polymeric combinations by the solvent evaporation technique. The physicochemical compatibility of the drug and the polymers was studied by infrared spectroscopy and differential scanning calorimetry. The results suggested no physicochemical incompatibility between the drug and the polymers. In vitro permeation studies were performed by using Franz diffusion cells. The results followed Higuchi kinetics (r = 0.9953-0.9979), and the mechanism of release was diffusion mediated. Based on physicochemical and in vitro skin permeation studies, patches coded as F3 (ethyl cellulose:polyvinylpyrrolidone, 7.5:2.5) and F6 (Eudragit RL:Eudragit RS, 8:2) were chosen for further in vivo studies. The bioavailability studies in rats indicated that the carvedilol transdermal patches provided steady state plasma concentrations with minimal fluctuations and improved bioavailability of 71% (for F3) and 62% (for F6) in comparison with oral administration. The antihypertensive activity of the patches in comparison with that of oral carvedilol was studied using methyl prednisolone acetate-induced hypertensive rats. It was observed that both the patches significantly controlled hypertension from the first hour (P < .05). The developed transdermal patches increase the efficacy of carvedilol for the therapy of hypertension. PMID- 17408219 TI - Influence of operational variables on properties of piroxicam pellets prepared by extrusion-spheronization: a technical note. PMID- 17408220 TI - Preparation and characterization of polyvinyl alcohol-gelatin hydrogel membranes for biomedical applications. AB - The purpose of this research was to design and develop hydrogels by esterification of polyvinyl alcohol (PVA) with gelatin. The membranes were characterized by Fourier Transform Infrared (FTIR) spectroscopy, x-ray diffraction (XRD), and differential scanning calorimetry. The viscosity of the esterified product (as solution) was compared with the mixture of PVA and gelatin of the same composition. The mechanical properties of the hydrogels were characterized by tensile tests. Swelling behavior and hemocompatibility of the membrane were also evaluated. The diffusion coefficient of salicylic acid (SA), when the receptor compartment contained Ringer's solution, through the membrane was determined. SA was used as a model drug. FTIR spectra of the membranes indicated complete esterification of the free carboxylic groups of gelatin. XRD studies indicated that the crystallinity of the membranes was mainly due to gelatin. The comparison of viscosity indicated an increase in segment density within the molecular coil. The membrane had sufficient strength and water-holding capacity. Hemocompatibility suggested that the hydrogel could be tried as wound dressing and as an implantable drug delivery system. The diffusion coefficient of SA through the membrane was found to be 1.32 x 10(-5) cm(2)/s. The experimental results indicated that the hydrogel could be tried for various biomedical applications. PMID- 17408221 TI - Formulation, evaluation, and comparison of bilayered and multilayered mucoadhesive buccal devices of propranolol hydrochloride. AB - The purpose of this research work was to establish mucoadhesive buccal devices of propranolol hydrochloride (PRH) in the forms of bilayered and multilayered tablets. The tablets were prepared using sodium carboxymethylcellulose (SCMC) and Carbopol-934 (CP) as bioadhesive polymers to impart mucoadhesion and ethyl cellulose (EC) to act as an impermeable backing layer. Buccal devices were evaluated by different parameters such as weight uniformity, content uniformity, thickness, hardness, surface pH, swelling index, ex vivo mucoadhesive strength, ex vivo mucoadhesion time, in vitro drug release, and in vitro drug permeation. As compared with bilayered tablets, multilayered tablets showed slow release rate of drug with improved ex vivo bioadhesive strength and enhanced ex vivo mucoadhesion time. The mechanism of drug release was found to be non-Fickian diffusion (value of n between 0.5 and 1.0) for both the buccal devices. The stability of drug in both the optimized buccal devices was tested for 6 hours in natural human saliva; both the buccal devices were found to be stable in natural human saliva. The present study concludes that mucoadhesive buccal devices of PRH can be a good way to bypass the extensive hepatic first-pass metabolism and to improve the bioavailability of PRH. PMID- 17408222 TI - Application of thermal effusivity as a process analytical technology tool for monitoring and control of the roller compaction process. AB - The aim of this study was to examine the relationship between physical characteristics of compacted ribbons and their thermal effusivity in an attempt to evaluate the feasibility of using effusivity for in-process monitoring of roller compaction. In this study, thermal effusivity, solid fraction, tensile strength, and Young's modulus of ribbons of microcrystalline cellulose (MCC), anhydrous lactose, and placebo (PBO) formulations containing various ratios of MCC to anhydrous lactose (75:20, 55:40, 40:55, and 20:75) were determined at various compaction pressures (25-150 bars). The effusivity-square root of solid fraction relationship was linear for MCC and all the PBO formulations but was a second-order polynomial function for lactose. This could be due to the predominant deformation of lactose by brittle fracture, which might have significantly increased the number and size of contact points between particles, causing a change in thermal conductivity along with a density change. The effusivity-tensile strength and effusivity-Young's modulus relationships were best described by logarithmic functions for MCC but were linear for lactose up to a compaction pressure of 65 bars. There were similar relationships for effusivity with tensile strength and Young's modulus for all PBO formulations except PBO IV, which might have been due to the deformation of lactose, the largest component in this formulation. Strong correlations between effusivity and physical properties of ribbons were established. Although these correlations were formulation dependent, they demonstrate the possibility of using effusivity as a tool in monitoring roller compaction. PMID- 17408223 TI - Preparation, characterization, and in vitro and in vivo evaluation of lovastatin solid lipid nanoparticles. AB - The purpose of this research was to study whether the bioavailability of lovastatin could be improved by administering lovastatin solid lipid nanoparticles (SLN) duodenally to rats. Lovastatin SLN were developed using triglycerides by hot homogenization followed by ultrasonication. Particle size and zeta potential were measured by photon correlation spectroscopy. The solid state of the drug in the SLN and lipid modification were characterized. Bioavailability studies were conducted in male Wistar rats after intraduodenal administration of lovastatin suspension and SLN. Stable lovastatin SLN having a mean size range of 60 to 119 nm and a zeta potential range of -16 to -21 mV were developed. More than 99% of the lovastatin was entrapped in the SLN. Lovastatin was dispersed in an amorphous state, and triglycerides were in beta(1) form in the SLN. In vitro stability studies showed the slow release and stability of lovastatin SLN. The relative bioavailabilities of lovastatin and lovastatin hydroxy acid of SLN were increased by ~173% and 324%, respectively, compared with the reference lovastatin suspension. PMID- 17408224 TI - Effect of selected non-ionic surfactants on the flow behavior of aqueous veegum suspensions. AB - The aim of this work was to investigate the influence of some non-ionic surfactants, Tween 80 and Brij 98, on the viscosity and flow behavior of a commercial montmorillonite clay, Veegum Granules. The effect of different concentrations of the surfactants on the shear stress-shear rate rheograms of hydrated concentrated clay suspensions was determined by shear viscometry. The addition of either surfactant increased the plastic viscosity and the yield stress of the suspensions. Furthermore, both surfactants altered the thixotropy of the suspensions to an extent that depended on both the surfactant concentration and the time of equilibration of the surfactant and Veegum. Brij 98 had a greater and more rapid effect. It is proposed that the surfactant polar head-groups anchor at the tetrahedral sheet surface, leaving the alkyl chains extending away from the edges and faces. Consequently, the alkyl chains undergo hydrophobic interactions that facilitate the association between the platelets and increase the physical structure within the suspension. Stereochemical differences between the polar groups may lead to differences in the way the surfactants associate with the tetrahedral sheet and hence their ultimate effect on the rheological behavior. There is a significant interaction between these surfactants and montmorillonite clays, and the rheological changes that occur could have a major impact on any pharmaceutical formulation that uses these ingredients. PMID- 17408225 TI - Development of a peptide-containing chewing gum as a sustained release antiplaque antimicrobial delivery system. AB - The objective of this study was to characterize the stability of KSL-W, an antimicrobial decapeptide shown to inhibit the growth of oral bacterial strains associated with caries development and plaque formation, and its potential as an antiplaque agent in a chewing gum formulation. KSL-W formulations with or without the commercial antibacterial agent cetylpyridinium chloride (CPC) were prepared. The release of KSL-W from the gums was assessed in vitro using a chewing gum apparatus and in vivo by a chew-out method. A reverse-phase high-performance liquid chromatography method was developed for assaying KSL-W. Raw material stability and temperature and pH effects on the stability of KSL-W solutions and interactions of KSL-W with tooth-like material, hydroxyapatite discs, were investigated. KSL-W was most stable in acidic aqueous solutions and underwent rapid hydrolysis in base. It was stable to enzymatic degradation in human saliva for 1 hour but was degraded by pancreatic serine proteases. KSL-W readily adsorbed to hydroxyapatite, suggesting that it will also adsorb to the teeth when delivered to the oral cavity. The inclusion of CPC caused a large increase in the rate and extent of KSL-W released from the gums. The gum formulations displayed promising in vitro/in vivo release profiles, wherein as much as 90% of the KSL-W was released in a sustained manner within 30 minutes in vivo. These results suggest that KSL-W possesses the stability, adsorption, and release characteristics necessary for local delivery to the oral cavity in a chewing gum formulation, thereby serving as a novel antiplaque agent. PMID- 17408226 TI - Comparison of a laboratory and a production coating spray gun with respect to scale-up. AB - A laboratory spray gun and a production spray gun were investigated in a scale-up study. Two Schlick spray guns, which are equipped with a new antibearding cap, were used in this study. The influence of the atomization air pressure, spray gun to tablet bed distance, polymer solution viscosity, and spray rate were analyzed in a statistical design of experiments. The 2 spray guns were compared with respect to the spray width and height, droplet size, droplet velocity, and spray density. The droplet size, velocity, and spray density were measured with a Phase Doppler Particle Analyzer. A successful scale-up of the atomization is accomplished if similar droplet sizes, droplet velocities, and spray densities are achieved in the production scale as in the laboratory scale. This study gives basic information for the scale-up of the settings from the laboratory spray gun to the production spray gun. Both spray guns are highly comparable with respect to the droplet size and velocity. The scale-up of the droplet size should be performed by an adjustment of the atomization air pressure. The scale-up of the droplet velocity should be performed by an adjustment of the spray gun to tablet bed distance. The presented statistical model and surface plots are convenient and powerful tools for scaling up the spray settings if the spray gun is changed from laboratory spray gun to the production spray gun. PMID- 17408227 TI - Product Quality Research Institute evaluation of cascade impactor profiles of pharmaceutical aerosols, part 1: background for a statistical method. AB - The purpose of this article is 2-fold: (1) to document in the public domain the considerations that led to the development of a regulatory statistical test for comparison of aerodynamic particle size distribution (APSD) of aerosolized drug formulations, which was proposed in a US Food and Drug Administration (FDA) draft guidance for industry; and (2) to explain the background and process for evaluation of that test through a working group involving scientists from the FDA, industry, academia, and the US Pharmacopeia, under the umbrella of the Product Quality Research Institute (PQRI). The article and the referenced additional statistical information posted on the PQRI Web site explain the reasoning and methods used in the development of the APSD test, which is one of the key tests required for demonstrating in vitro equivalence of orally inhaled and nasal aerosol drug products. The article also describes the process by which stakeholders with different perspectives have worked collaboratively to evaluate properties of the test by drawing on statistical models, historical and practical information, and scientific reasoning. Overall, this article provides background information to accompany the companion article's discussion of the study's methods and results. PMID- 17408228 TI - Product Quality Research Institute evaluation of cascade impactor profiles of pharmaceutical aerosols: part 2--evaluation of a method for determining equivalence. AB - The purpose of this article is to present the thought process, methods, and interim results of a PQRI Working Group, which was charged with evaluating the chi-square ratio test as a potential method for determining in vitro equivalence of aerodynamic particle size distribution (APSD) profiles obtained from cascade impactor measurements. Because this test was designed with the intention of being used as a tool in regulatory review of drug applications, the capability of the test to detect differences in APSD profiles correctly and consistently was evaluated in a systematic way across a designed space of possible profiles. To establish a "base line," properties of the test in the simplest case of pairs of identical profiles were studied. Next, the test's performance was studied with pairs of profiles, where some difference was simulated in a systematic way on a single deposition site using realistic product profiles. The results obtained in these studies, which are presented in detail here, suggest that the chi-square ratio test in itself is not sufficient to determine equivalence of particle size distributions. This article, therefore, introduces the proposal to combine the chi-square ratio test with a test for impactor-sized mass based on Population Bioequivalence and describes methods for evaluating discrimination capabilities of the combined test. The approaches and results described in this article elucidate some of the capabilities and limitations of the original chi-square ratio test and provide rationale for development of additional tests capable of comparing APSD profiles of pharmaceutical aerosols. PMID- 17408229 TI - Formulation development and in vitro and in vivo evaluation of membrane-moderated transdermal systems of ampicillin sodium in ethanol: pH 4.7 buffer solvent system. AB - The objective of the present study was to develop membrane-moderated transdermal systems of ampicillin sodium and to evaluate them with respect to various in vitro and in vivo parameters. The membrane-type transdermal systems were prepared using a drug with various antinucleant polymers-hydroxypropyl methylcellulose (HPMC), methylcellulose (MC), cellulose acetate phthalate, chitosan, sodium alginate (SA), and sodium carboxymethylcellulose-in an ethanol: pH 4.7 buffer volatile system by the solvent evaporation technique with HPMC as the rate controlling membrane for all the systems. The swelling properties of the polymers were studied, and drug-polymer interaction studies were performed. The patches were subjected to various physicochemical studies, in vitro release studies, permeation studies, and skin irritation studies. The best patch among the formulations was selected for further in vivo studies. Compared to the other patches, SA exhibited the highest moisture content at 16%; a 21% moisture uptake was found with MC. The release and permeation of the drug from the SA patch was found to be the maximum. The in vivo study of the SA patch exhibited a peak plasma concentration C(max) of 126 microg/mL at T(max) 4 hours. Hence, it can be concluded that hydrophilic ampicillin sodium can be developed as a transdermal delivery system with SA that is an alternative to intravenous administration and has minimal adverse effects. PMID- 17408230 TI - Evaluation of microcrystalline cellulose prepared from sisal fibers as a tablet excipient: a technical note. PMID- 17408231 TI - Preparation and assessment of novel coprocessed superdisintegrant consisting of crospovidone and sodium starch glycolate: a technical note. PMID- 17408233 TI - Biomarker qualification pilot process at the US Food and Drug Administration. AB - New biomarkers of safety and efficacy are becoming powerful tools in drug development. Their application can be accelerated if a consensus can be reached about their qualification for regulatory applications. This consensus requires a review structure within the US Food and Drug Administration (FDA) that can evaluate qualification data for these biomarkers and determine whether these biomarkers can be qualified. A pilot process and corresponding Biomarker Qualification Review Team have been developed to test how the FDA can work on biomarker qualification. PMID- 17408232 TI - Dual dopamine/serotonin releasers as potential medications for stimulant and alcohol addictions. AB - We have advocated the idea of agonist therapy for treating cocaine addiction. This strategy involves administration of stimulant-like medications (eg, monoamine releasers) to alleviate withdrawal symptoms and prevent relapse. A major limitation of this approach is that many candidate medicines possess significant abuse potential because of activation of mesolimbic dopamine (DA) neurons in central nervous system reward circuits. Previous data suggest that serotonin (5-HT) neurons can provide an inhibitory influence over mesolimbic DA neurons. Thus, it might be predicted that the balance between DA and 5-HT transmission is important to consider when developing medications with reduced stimulant side effects. In this article, we discuss several issues related to the development of dual DA/5-HT releasers for the treatment of substance use disorders. First, we discuss evidence supporting the existence of a dual deficit in DA and 5-HT function during withdrawal from chronic cocaine or alcohol abuse. Then we summarize studies that have tested the hypothesis that 5-HT neurons can dampen the effects mediated by mesolimbic DA. For example, it has been shown that pharmacological manipulations that increase extracellular 5-HT attenuate stimulant effects produced by DA release, such as locomotor stimulation and self administration behavior. Finally, we discuss our recently published data about PAL-287 (naphthylisopropylamine), a novel non-amphetamine DA-/5-HT-releasing agent that suppresses cocaine self-administration but lacks positive reinforcing properties. It is concluded that DA/5-HT releasers might be useful therapeutic adjuncts for the treatment of cocaine and alcohol addiction, obesity, and even attention deficit disorder and depression. PMID- 17408234 TI - Key elements of bioanalytical method validation for small molecules. AB - Method validation is a process that demonstrates that a method will successfully meet or exceed the minimum standards recommended in the Food and Drug Administration (FDA) guidance for accuracy, precision, selectivity, sensitivity, reproducibility, and stability. This article discusses the validation of bioanalytical methods for small molecules with emphasis on chromatographic techniques. We present current thinking on validation requirements as described in the current FDA Guidance and subsequent 2006 Bioanalytical Methods Validation Workshop white paper. PMID- 17408235 TI - Vectors for airway gene delivery. AB - Delivery of genes to the airway epithelium for therapeutic purposes seemed easy at first, because the epithelial cells interface with the environment and are therefore accessible. However, problems encountered were more substantial than were originally expected. Nonviral systems may be preferred for long-term gene expression, for they can be dosed repeatedly. Two nonviral gene transfer systems have been in clinical trials, lipid-mediated gene transfer and DNA nanoparticles. Both have sufficient efficiency to be candidates for correction of the cystic fibrosis defect, and both can be dosed repeatedly. However, lipid-mediated gene transfer in the first generation provokes significant inflammatory toxicity, which may be engineered out by adjustments of the lipids, the plasmid CpG content, or both. Both lipid-mediated gene transfer and DNA nanoparticles in the first generation have short duration of expression, but reengineering of the plasmid DNA to contain mostly eukaryotic sequences may address this problem. Considerable advances in the understanding of the cellular uptake and expression of these agents and in their practical utility have occurred in the last few years; these advances are reviewed here. PMID- 17408236 TI - Peptide-guided gene delivery. AB - Although currently less efficient than their viral counterparts, nonviral vectors are under intense investigation as a safer alternative for gene therapy. For successful delivery, the nonviral vector must be able to overcome many barriers to protect DNA and specifically deliver it for efficient gene expression in target cells. The use of peptides as gene delivery vectors is advantageous over other nonviral agents in that they are able to achieve all of these goals. This review will focus on the application of peptides to mediate nonviral gene delivery. By examining the literature over the past 20 years, it becomes clear that no other class of biomolecules are simultaneously capable of DNA condensation, blocking metabolism, endosomal escape, nuclear localization, and receptor targeting. Based on virtually limitless diversity of peptide sequence and function information from nature, it is increasingly clear that peptide guided gene delivery is still in its infancy. PMID- 17408237 TI - A survey of population analysis methods and software for complex pharmacokinetic and pharmacodynamic models with examples. AB - An overview is provided of the present population analysis methods and an assessment of which software packages are most appropriate for various PK/PD modeling problems. Four PK/PD example problems were solved using the programs NONMEM VI beta version, PDx-MCPEM, S-ADAPT, MONOLIX, and WinBUGS, informally assessed for reasonable accuracy and stability in analyzing these problems. Also, for each program we describe their general interface, ease of use, and abilities. We conclude with discussing which algorithms and software are most suitable for which types of PK/PD problems. NONMEM FO method is accurate and fast with 2 compartment models, if intra-individual and interindividual variances are small. The NONMEM FOCE method is slower than FO, but gives accurate population values regardless of size of intra- and interindividual errors. However, if data are very sparse, the NONMEM FOCE method can lead to inaccurate values, while the Laplace method can provide more accurate results. The exact EM methods (performed using S-ADAPT, PDx-MCPEM, and MONOLIX) have greater stability in analyzing complex PK/PD models, and can provide accurate results with sparse or rich data. MCPEM methods perform more slowly than NONMEM FOCE for simple models, but perform more quickly and stably than NONMEM FOCE for complex models. WinBUGS provides accurate assessments of the population parameters, standard errors and 95% confidence intervals for all examples. Like the MCPEM methods, WinBUGS's efficiency increases relative to NONMEM when solving the complex PK/PD models. PMID- 17408238 TI - Informatics: the fuel for pharmacometric analysis. AB - The current informal practice of pharmacometrics as a combination art and science makes it hard to appreciate the role that informatics can and should play in the future of the discipline and to comprehend the gaps that exist because of its absence. The development of pharmacometric informatics has important implications for expediting decision making and for improving the reliability of decisions made in model-based development. We argue that well-defined informatics for pharmacometrics can lead to much needed improvements in the efficiency, effectiveness, and reliability of the pharmacometrics process. The purpose of this paper is to provide a description of the pervasive yet often poorly appreciated role of informatics in improving the process of data assembly, a critical task in the delivery of pharmacometric analysis results. First, we provide a brief description of the pharmacometric analysis process. Second, we describe the business processes required to create analysis-ready data sets for the pharmacometrician. Third, we describe selected informatic elements required to support the pharmacometrics and data assembly processes. Finally, we offer specific suggestions for performing a systematic analysis of existing challenges as an approach to defining the next generation of pharmacometric informatics. PMID- 17408240 TI - High-throughput system for analyzing ligand-induced cofactor recruitment by vitamin D receptor. AB - A high-throughput screening system for analyzing small molecule-induced coactivator (CoA) recruitment by the vitamin D receptor (VDR) has been developed. The vitamin D-induced protein-protein interactions between VDR and fluorophore (Cy3 or Cy5)-labeled TIF2 or SRC-1 were successfully detected by using a new HCHO fixing method of the protein complex on microplates. The results obtained from this screening of our synthetic vitamin D analogues suggest that the CoA recruiting activities play an important role in determining the biological activity of various vitamin D analogues and explain the discrepancies between the VDR binding affinity and their biological activity. PMID- 17408239 TI - Nonviral gene delivery: what we know and what is next. AB - Gene delivery using nonviral approaches has been extensively studied as a basic tool for intracellular gene transfer and gene therapy. In the past, the primary focus has been on application of physical, chemical, and biological principles to development of a safe and efficient method that delivers a transgene into target cells for appropriate expression. This review summarizes the current status of the most commonly used nonviral methods, with an emphasis on their mechanism of action for gene delivery, and their advantages and limitations for gene therapy applications. The technical aspects of each delivery system are also reviewed, with a focus on how to achieve optimal delivery efficiency. A brief discussion of future development and further improvement of the current systems is intended to stimulate new ideas and encourage rapid advancement in this new and promising field. PMID- 17408241 TI - Novel approach for the numerical characterization of molecular chirality. AB - The use of chiral compounds as pharmaceuticals and agrochemicals continues to increase, warranting numerical characterization of chirality in order to develop structure-activity relationship models involving these compounds. Enantiomers are identical in all scalar properties and, hence, are not differentiated by topological indices and 3-D descriptors. Three distinct measures of chirality were developed to discriminate diastereomers and enantiomers. The novel topological indices treat chirality as a continuous measure, and hence we prefer to call it the Relative Chirality Index (RCI). Application of RCI in developing SAR is illustrated with the repellency data for the diastereomers of picaridin and AI3-37220. PMID- 17408242 TI - Enzymatic spectrophotometric method for aflatoxin B detection based on acetylcholinesterase inhibition. AB - A new method for aflatoxin B (AFB) determination is proposed. The AFB determination is based on acetylcholinesterase (AChE) inhibition, and the AChE residual activity is determined using the colorimetric method (Ellman's method). Cholinesterases (ChEs) from various sources were tested using AFB1 as reference aflatoxin. AChE from electric eel has shown the highest sensitivity to AFB1, and it was chosen for the rest of the work. To select and optimize the analytical procedures, an investigation on the type of AChE inhibition by AFB1 was carried out. The AChE degree of inhibition by AFB1 was independent of the incubation time and the enzyme concentrations, showing the reversibility of the inhibition. This reversibility of the inhibition permits a rapid analysis of AFB1, requiring only 3 min. For the development of the AFB1 assay, the pH, the time of reaction, temperature, and substrate concentration were evaluated and optimized. The linear range of 10-60 ng mL-1 was determined. To evaluate the selectivity of this method, the cross-reactivity with other aflatoxins such as aflatoxin B2, aflatoxin G1, aflatoxin G2, and aflatoxin M1 was investigated. Finally, the suitability of the assay for AFB1 quantification in barley was evaluated. This study shows a new approach to detect aflatoxins based on enzyme inhibition and has advantages such as the ease of use, rapidity, and cost effectiveness. Thus, it could find a possible use as a screening method for this type of mycotoxins. PMID- 17408243 TI - Untargeted analysis of mass spectrometry data for elucidation of metabolites and function of enzymes. AB - A Matlab-based computer program termed Discovery of General Endo- and Xenobiotics (DoGEX) was developed, which uses wavelets and morphological analysis to process liquid chromatography-mass spectrometry (LC-MS) data. The output of the program is a list of integration areas as a function of retention time and molecular mass. A feature of the computer program is spectral filtering to facilitate the detection of chromatographic peaks with a particular isotopic ratio. The program DoGEX was used to automatically select oxidation products formed from felodipine (i.e., two chlorine atoms) and bromocriptine (one bromine atom) with cytochrome P450 3A4. The recognized isotope ratio can be changed to permit a natural or artificial mixture of isotopes to be monitored for selections. This computer program can be used to analyze LC-MS data for untargeted metabolic profiling experiments, e.g., to assign endogenous functions to newly characterized cytochrome P450 enzymes. In a representative example, an incubation of testosterone, NADPH, and a 1:1 16O2/18O2 mixture yielded products with M and M+2 ions resembling bromine doublets. Another use of the program is the subtraction of one set of tR, m/z data from another, e.g., in comparisons of changes in patterns during enzyme reactions. PMID- 17408244 TI - Elution-extrusion countercurrent chromatography: theory and concepts in metabolic analysis. AB - Elution-extrusion countercurrent chromatography (EECCC) takes full advantage of the liquid nature of the stationary phase in CCC by combining regular chromatographic elution with stationary-phase extrusion. EECCC is shown to be a three-stage process consisting of classical elution (I), sweeping elution (II), and extrusion (III). After only two column volumes of solvent, it rapidly yields a high-resolution chromatogram that covers an extended polarity range of solutes. As hydrophilicity/lipophilicity balance is a crucial discriminatory property of analytes in highly complex mixtures such as metabolomic samples, the precise determination of CCC distribution constants (KD) is vital to the analysis of metabolomes and other complex biological matrixes. This work builds the EECCC concept by performing a full theoretical treatment and providing equations for retention volumes, peak widths, resolution factors, and distribution constants. Experimental validation utilizes natural products standards that resemble the zero to infinity range of the polarity continuum. EECCC extends the "sweet spot" of high resolution in CCC and provides access to the otherwise practically unapproachable high-KD portion of the high-resolution chromatograms in CCC. Its improved capabilities of KD targeting make EECCC a promising tool for the specific analysis of "small" molecules in complex samples such as in metabolomic fingerprinting and footprinting. PMID- 17408245 TI - Monitoring cell adhesion by piezoresonators: impact of increasing oscillation amplitudes. AB - In recent years, the quartz crystal microbalance (QCM) has been established as a sensitive analytical tool to monitor the attachment and spreading of mammalian cells to in vitro surfaces. Due to its superior time resolution, the device is capable of reading even subtle differences in cell adhesion kinetics. However, thickness shear mode piezoresonators, which are the core component of the QCM approach, can be used not only as a sensor but also as an actuator when the oscillation amplitude of the crystal is increased so that molecular recognition at the solid-liquid interface is disturbed. In this study, we have addressed the impact of elevated lateral oscillation amplitudes on the adhesion kinetics of three mammalian cell lines. We used AT-cut piezoresonators with a fundamental resonance frequency of 5 MHz, and the analytical readout was performed by impedance analysis. Formation of stable cell-substrate contacts is retarded or entirely blocked when the lateral oscillation amplitude (in the center of the resonator) exceeds values higher than 20 nm. Shear oscillations of similar amplitude were, however, not sufficient to displace attached cells from the surface. Moreover, the experimental data prove that the normal QCM readout with oscillation amplitudes smaller than 1 nm is, indeed, non-invasive with respect to mammalian cells. PMID- 17408246 TI - Thermodynamic characterization of the protein-protein interaction in the heteromeric Bacillus subtilis pyridoxalphosphate synthase. AB - Two biosynthetic routes for the synthesis of pyridoxal 5'-phosphate (PLP), the biologically active compound of vitamin B6, have been characterized. The major pathway leads to direct formation of PLP from a pentasaccharide and a trisaccharide and is operative in plants, fungi, protozoa, and bacteria. This reaction is catalyzed by a single glutamine amidotransferase enzyme complex consisting of a pyridoxal synthase, termed Pdx1, and a glutaminase, termed Pdx2. In this complex, Pdx2 generates ammonia from L-glutamine and supplies it to Pdx1 for incorporation into PLP. The glutaminase activity of Pdx2 requires the presence of Pdx1 in a heteromeric complex, previously characterized by a crystallographic three-dimensional (3D) structure determination. Here, we give a thermodynamic description of complex formation of Bacillus subtilis PLP synthase in the absence or presence of L-glutamine. We show that L-glutamine directly affects the tightness of the protein complex, which exhibits dissociation constants of 6.9 and 0.3 microM in its absence and presence, respectively (at 25 degrees C). This result relates to the positioning of L-glutamine on the heterodimer interface as seen in the 3D structure. In an analysis of the temperature dependence of the enthalpy, negative heat capacity changes (deltaCp) agree with a protein interface governed by hydrophobic interactions. The measured heat capacity change is also a function of L-glutamine, with a negative deltaCp in the presence of L-glutamine and a more negative one in its absence. These findings suggest that L-glutamine not only affects the strength of complex formation but also determines the forces involved in complex formation, with regard to different relative contributions of hydrophobic and hydrophilic interactions. PMID- 17408247 TI - The reaction of n- and i-C4H5 radicals with acetylene. AB - In this article, we discuss the reactions of i-C4H5 and n-C4H5 with acetylene. Both have been proposed as possible cyclization steps, forming benzene or fulvene, in rich flames burning aliphatic fuels. The relevant parts of the potential energy surface were determined from rQCISD(T) calculations extrapolated to the infinite-basis-set limit. Using this information in a Rice-Ramsperger Kassel-Marcus-based master equation, we have calculated thermal rate coefficients and product distributions for both reactions as a function of temperature and pressure. The results are cast in forms that can be used in modeling, and the implications of the results for flame chemistry are discussed. PMID- 17408248 TI - Effect of the diamine nonleaving group in platinum-acridinylthiourea conjugates on DNA damage and cytotoxicity. AB - The following complexes of type [PtCl(R)(ACRAMTU)](NO3)2 (ACRAMTU = 1-[2-(acridin 9-ylamino)ethyl]-1,3-dimethylthiourea)), derived from prototype 1 (with R = ethane-1,2-diamine), were synthesized: 2 (with R = (1R,2R)-1,2 diaminocyclohexane), 3 (with R = propane-1,3-diamine), 4 (with R = N1,N1,N2,N2 tetramethylethane-1,2-diamine), and 5 (with R = 2,2'-bipyridine). The DNA sequence specificity of the conjugates and their antiproliferative potential in HL-60 and H460 cells were investigated. Conjugate 3 showed the strongest non cisplatin-type DNA damage in polymerase stop assays and superior cell kill efficacy in H460 lung cancer (IC50 = 70 nM). PMID- 17408250 TI - Proteomics of neuroendocrine secretory vesicles reveal distinct functional systems for biosynthesis and exocytosis of peptide hormones and neurotransmitters. AB - Regulated secretory vesicles produce, store, and secrete active peptide hormones and neurotransmitters that function in cell-cell communication. To gain knowledge of the protein systems involved in such secretory vesicle functions, we analyzed proteins in the soluble and membrane fractions of dense core secretory vesicles purified from neuroendocrine chromaffin cells. Soluble and membrane fractions of these vesicles were subjected to SDS-PAGE separation, and proteins from systematically sectioned gel lanes were identified by microcapillary LC-MS/MS (microLC-MS/MS) of tryptic peptides. The identified proteins revealed functional categories of prohormones, proteases, catecholamine neurotransmitter metabolism, protein folding, redox regulation, ATPases, calcium regulation, signaling components, exocytotic mechanisms, and related functions. Several novel secretory vesicle components involved in proteolysis were identified consisting of cathepsin B, cathepsin D, cystatin C, ubiquitin, and TIMP, as well carboxypeptidase E/H and proprotein convertases that are known to participate in prohormone processing. Significantly, the membrane fraction exclusively contained an extensive number of GTP nucleotide-binding proteins related to Rab, Rho, and Ras signaling molecules, together with SNARE-related proteins and annexins that are involved in trafficking and exocytosis of secretory vesicle components. Membranes also preferentially contained ATPases that regulate proton translocation. These results implicate membrane-specific functions for signaling and exocytosis that allow these secretory vesicles to produce, store, and secrete active peptide hormones and neurotransmitters released from adrenal medulla for the control of physiological functions in health and disease. In summary, this proteomic study illustrates secretory vesicle protein systems utilized for the production and secretion of regulatory factors that control neuroendocrine functions. PMID- 17408249 TI - Potent nonpeptide antagonists of the bradykinin B1 receptor: structure-activity relationship studies with novel diaminochroman carboxamides. AB - The bradykinin B1 receptor is induced following tissue injury and/or inflammation. Antagonists of this receptor have been studied as promising candidates for treatment of chronic pain. We have identified aryl sulfonamides containing a chiral chroman diamine moiety that are potent antagonists of the human B1 receptor. Our previously communicated lead, compound 2, served as a proof-of-concept molecule, but suffered from poor pharmacokinetic properties. With guidance from metabolic profiling, we performed structure-activity relationship studies and have identified potent analogs of 2. Variation of the sulfonamide moiety revealed a preference for 3- and 3,4-disubstituted aryl sulfonamides, while bulky secondary and tertiary amines were preferred at the benzylic amine position for potency at the B1 receptor. Modifying the beta-amino acid core of the molecule lead to the discovery of highly potent compounds with improved in vitro pharmacokinetic properties. The most potent analog at the human receptor, compound 38, was also active in a rabbit B1 receptor cellular assay. Furthermore, compound 38 displayed in vivo activity in two rabbit models, a pharmacodynamic model with a blood pressure readout and an efficacy model of inflammatory pain. PMID- 17408251 TI - Tuning J-aggregates of tetra(p-hydroxyphenyl)porphyrin by the headgroups of ionic surfactants in acidic nonionic micellar solution. AB - J-aggregates of the diacid form of tetra(p-hydroxyphenyl)porphyrin (THPP) were found to be stable in nonionic micellar solution in the presence of trace ionic surfactant with an oxyacid headgroup. The excitation energy of exciton coupling depends systematically on the headgroups of the ionic surfactant, by which strong and weak coupling can be accomplished in the J-aggregates. The J-aggregates have two strong exciton bands corresponding to the B- and Q-bands of the protonated monomers. The total fluorescence of THPP is quenched through aggregate formation. A strong and sharply peaked resonance light-scattering signal that suggests a delocalized excitonic state was observed just slightly to the red of the absorption maximum of the J-aggregates. The overall resonance Raman intensities appeared to be stronger in the aggregates than in the monomers. In the kinetics of aggregation induced by sodium dodecyl sulfate (SDS), no characteristics of autocatalyzed reactions were observed, and there was only a logarithmic phase that lasted only several seconds. PMID- 17408252 TI - Ab initio molecular dynamics study of carbon dioxide and bicarbonate hydration and the nucleophilic attack of hydroxide on CO2. AB - We apply ab initio molecular dynamics (AIMD) to study the hydration structures of the carbon dioxide molecule and the bicarbonate and carbonate anions in liquid water. We also compute the free energy change associated with the nucleophilic attack of the hydroxide ion on carbon dioxide. CO2 behaves like a hydrophobic species and exhibits weak interactions with water molecules. The bicarbonate and carbonate ions are strongly hydrated and coordinate to an average of 6.9 and 8.7 water molecules, respectively. The energetics for the reaction in the gas phase are investigated using density functional theory and second-order Moller-Plesset perturbation theory (MP2) in conjunction with high-quality basis sets. Using umbrella sampling techniques, we compute the standard state, aqueous phase free energy difference associated with the reaction CO2+OH--->HCO3- after correcting AIMD energies with MP2 results. Our predictions are in good agreement with experiments. The hydration structures along the reaction coordinate, which give rise to a predicted 9.7 kcal/mol standard state free energy barrier, are further analyzed. PMID- 17408253 TI - Are hydroxyl-containing biomolecules important in biosilicification? A model study. AB - To understand the roles of hydroxyl-containing biomolecules in biosilicification, theoretical and experimental studies of silica formation utilizing biological and model organic additives have been undertaken. However, the role of hydroxyl functionalized biomolecules in silica formation is still not fully understood. To address this problem, we performed a systematic in vitro study of silica formation in the presence of two proteins rich in hydroxyl-containing amino acids (native sericin proteins extracted from Bombyx mori and a recombinant sericin precursor peptide) and a range of small alkanediols. The data obtained suggest the following hypotheses for the role of hydroxyl-containing organic molecules in silica formation. In the first case, hydroxyl-containing organic molecules are not at all involved chemically in the formation of silica. Instead, they may be only assisting in rendering stability and solubility to the organic molecules found occluded in silica. The second possibility is that if we assume that hydroxyl-containing organic molecules affect silica formation, then the environments of silicic acid polymerization could be highly deficient in water to increase the effects of hydroxyl functional groups of proteins in silica formation. These results and their implications are discussed in the context of biosilicification and biomineralization. PMID- 17408254 TI - An anisotropic united atoms (AUA) potential for thiophenes. AB - The optimization of the parameters of the Lennard-Jones (LJ) 12-6 potential of the sulfur atom in thiophene has allowed the AUA 4 potential to be successfully extended to alkyl and polythiophenes. Monte Carlo Gibbs ensemble and grand canonical simulations combined with histogram reweighting techniques have been performed to investigate the resulting phase equilibrium and the critical region of different molecules of this family in order to test the proposed potential. Excellent agreement with experimental densities, enthalpies of vaporization, and saturation pressures has been obtained in most of the cases. In particular, the critical point of our model for thiophene has been located with a statistical precision of less than 0.1% and is within 1% of the experimental value. The calculation of the critical points has been made through a recently implemented methodology based on the calculation of a fourth order cumulant (Binder parameter) combined with the use of finite size scaling methods, allowing the critical points to be located in a straightforward and accurate way. PMID- 17408255 TI - Effect of temperature and water content on the shear viscosity of the ionic liquid 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide as studied by atomistic simulations. AB - Atomistic simulations are conducted to examine the dependence of the viscosity of 1-ethyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide on temperature and water content. A nonequilibrium molecular dynamics procedure is utilized along with an established fixed charge force field. It is found that the simulations quantitatively capture the temperature dependence of the viscosity as well as the drop in viscosity that occurs with increasing water content. Using mixture viscosity models, we show that the relative drop in viscosity with water content is actually less than that that would be predicted for an ideal system. This finding is at odds with the popular notion that small amounts of water cause an unusually large drop in the viscosity of ionic liquids. The simulations suggest that, due to preferential association of water with anions and the formation of water clusters, the excess molar volume is negative. This means that dissolved water is actually less effective at lowering the viscosity of these mixtures when compared to a solute obeying ideal mixing behavior. The use of a nonequilibrium simulation technique enables diffusive behavior to be observed on the time scale of the simulations, and standard equilibrium molecular dynamics resulted in sub diffusive behavior even over 2 ns of simulation time. PMID- 17408256 TI - Probing intermolecular interactions in water/ionic liquid mixtures by far infrared spectroscopy. AB - Far-infrared spectra in the range from 600 to 20 cm-1 of two hydrophilic (1-ethyl 3-methylimidazolium tetrafluoroborate and 1-butyl-3-methylimidazolium tetrafluoroborate) and one hydrophobic (1-butyl-3-methylimidazolium hexafluorophosphate) ionic liquids and their mixtures with water at different concentrations are reported. Shifts of the librational water bands depending on the nature of the anion are found to be related to the strength of the interaction between the water molecules and the anions. For both hydrophilic ionic liquids, the librational band is centered around 460 cm-1, whereas for the hydrophobic ionic liquid, it is shifted to 388 cm-1, indicating less hindered rotation of single water molecules. Multivariate curve resolution, paying special attention to the spectral range from 50 to 350 cm-1, was used to investigate the presence of different species with increasing water concentration. For both hydrophilic ionic liquids, a band located at 153 cm-1 was resolved into two different contributions. A small contribution at 202 cm-1 can be attributed to intermolecular interactions between water molecules forming dimers. The major contribution (centered at 148 cm-1) corresponds to water molecules that do not bond to each other via H-bonding. It is therefore assigned to a hindered translation arising from the stretching of the hydrogen bond between BF4- anions and water molecules. Formation of water dimers in the hydrophobic ionic liquid does not occur. Furthermore, the spectral contribution of the stretching of H bonds between water molecules and PF6- cannot be unambiguously detected, which indicates an extremely weak interaction between water molecules and this anion. PMID- 17408257 TI - Mechanisms for fluorescence depolarization in dendrimers. AB - We have investigated the fluorescence properties of dendrimers (Gn is the dendrimer generation number) containing four different luminophores, namely terphenyl (T), dansyl (D), stilbenyl (S), and eosin (E). In the case of T, the dendrimers contain a single p-terphenyl fluorescent unit as a core with appended sulfonimide branches of different size and n-octyl chains. In the cases of D and S, multiple fluorescent units are appended in the periphery of poly(propylene amine) dendritic structures. In the case of E, the investigated luminophore is noncovalently linked to the dendritic scaffold, but is encapsulated in cavities of a low luminescent dendrimer. Depending on the photophysical properties of the fluorescent units and the structures of the dendrimers, different mechanisms of fluorescence depolarization have been observed: (i) global rotation for GnT dendrimers; (ii) global rotation and local motions of the dansyl units at the periphery of GnD dendrimers; (iii) energy migration among stylbenyl units in G2S; and (iv) restricted motion when E is encapsulated inside a dendrimer, coupled to energy migration if the dendrimer hosts more than one eosin molecule. PMID- 17408258 TI - Computational studies of electron paramagnetic resonance parameters for paramagnetic molybdenum complexes. 1. Method validation on small and medium-sized systems. AB - A variety of density functional methods have been evaluated in the computation of electronic g-tensors and molybdenum hyperfine couplings for systems ranging from the Mo atom through MoIIIN, [MoVOCl4]-, and [MoVOF5]2- to two larger MoV complexes MoXLCl2 (X=O, S; L=tris(3,5-dimethylpyrazolyl)hydroborate anion). In particular, the influence of the molybdenum basis set and of various exchange correlation functionals with variable admixtures of Hartree-Fock exchange on the computed EPR parameters have been evaluated in detail. Careful basis-set studies have provided a moderate-sized 12s6p5d all-electron basis on molybdenum that gives hyperfine tensors in excellent agreement with much larger basis sets and that will be useful for calculations on larger systems. The best agreement with experimental data for both hyperfine and g-tensors is obtained with hybrid functionals containing approximately 30-40% Hartree-Fock exchange. Only for MoSLCl2 does increasing spin contamination with increasing exact-exchange admixture restrict the achievable computational accuracy. In all cases, spin orbit corrections to the hyperfine tensors are sizable and have to be included in accurate calculations. Scalar relativistic effects enhance the isotropic Mo hyperfine coupling by approximately 15-20%. Two-component g-tensor calculations with variational inclusion of spin-orbit coupling show that the Deltag parallel components in [MoVOCl4]- and [MoVOF5]2- depend on higher-order spin-orbit contributions and are thus described insufficiently by the usual second-order perturbation approaches. Computed orientations of g- and hyperfine tensors relative to each other and to the molecular framework for the MoXLCl2 complexes provide good agreement between theory and single-crystal electron paramagnetic resonance experiments. In these cases, the hyperfine tensor orientations are influenced only slightly by spin-orbit effects. PMID- 17408259 TI - Pressure-induced phase transitions in PbTiO3: a query for the polarization rotation theory. AB - Our first-principles computations show that the ground state of PbTiO3 under hydrostatic pressure transforms discontinuously from P4mm to R3c at 9 GPa. Spontaneous polarization decreases with increasing pressure so that the R3c phase transforms to the centrosymmetric Rc phase at around 27 GPa. The first-order phase transition between the tetragonal and rhombohedral phases is exceptional since there is no evidence for a bridging phase. The essential feature of the R3c and Rc phases is that they allow the oxygen octahedron to increase its volume VB at the expense of the cuboctahedral volume VA around a Pb ion. This is further supported by the fact that neither the R3m nor Cm phase, which keep the VA/VB ratio constant, is a ground state within the pressure range between 0 and 40 GPa. Thus, tetragonal strain is dominant up to 9 GPa, whereas at higher pressures, efficient compression through oxygen octahedra tilting plays the central role for PbTiO3. Previously predicted pressure induced colossal enhancement of piezoelectricity in PbTiO3 corresponds to unstable Cm and R3m phases. This suggests that the phase instability, in contrast to the polarization rotation, is responsible for the large piezoelectric properties observed in systems like Pb(Zr,Ti)O3 in the vicinity of the morphotropic phase boundary. PMID- 17408260 TI - Wetting and electrowetting properties of carbon nanotube templated parylene films. AB - In this study, we compared the wetting and electrowetting properties of a planar parylene (poly(p-xylylene)) film to those of a nanostructured parylene film. To generate the nanostructured film, we used an aligned array of multiwalled carbon nanotubes as a template; a thin coating of parylene was deposited on the nanotube template to generate a parylene film with a nanoscale roughness structure. Static contact angle measurements indicated a very significant increase in the water contact angle from approximately 73 degrees for planar parylene to approximately 110 degrees for the nanotemplated parylene. In addition, we performed electrowetting experiments to dynamically tune the contact angle by application of electric potential. Interestingly, the flat parylene film showed contact angle saturation at an applied voltage of approximately 40 V, while the nanotemplated parylene film did not experience saturation in the contact angle response even for voltages up to 80 V. These results show that engineering a nanoscale roughness structure to a polymer film results in significant changes to the wetting and electrowetting properties of the polymer. PMID- 17408261 TI - Series of trinuclear NiIILnIIINiII complexes derived from 2,6 Di(acetoacetyl)pyridine: synthesis, structure, and magnetism. AB - Eighteen trinuclear NiII2LnIII complexes of 2,6-di(acetoacetyl)pyridine (H2L) (Ln=La-Lu except for Pm) were prepared by a "one-pot reaction" of H2L, Ni(NO3)2.6H2O, and Ln(NO3)3.nH2O in methanol. X-ray crystallographic studies indicate that two L2- ligands sandwich two NiII ions with the terminal 1,3 diketonate sites and one LnIII ion with the central 2,6-diacylpyridine site, forming the trinuclear [Ni2Ln(L)2] core of a linear NiLnNi structure. The terminal Ni assumes a six-coordinate geometry together with methanol or water molecules, and the central Ln assumes a 10-coordinate geometry together with two or three nitrate ions. The [Ni2Ln(L)2] core is essentially coplanar for large Ln ions (La, Ce, Pr, Nd) but shows a distortion with respect to the two L2- ligands for smaller Ln ions. Magnetic studies for the Ni2Ln complexes of diamagnetic LaIII and LuIII indicate an antiferromagnetic interaction between the terminal NiII ions. A magnetic analysis of the Ni2Gd complex based on the isotropic Heisenberg model indicates a ferromagnetic interaction between the adjacent NiII and GdIII ions and an antiferromagnetic interaction between the terminal NiII ions. The magnetic properties of other Ni2Ln complexes were studied on the basis of a numerical approach with the Ni2La complex and analogous Zn2Ln complexes, and they indicated that the NiII-LnIII interaction is weakly antiferromagnetic for Ln=Ce, Pr, and Nd and ferromagnetic for Ln=Gd, Tb, Dy, Ho, and Er. PMID- 17408262 TI - The origin of the surprising stabilities of highly charged self-assembled polymetallic complexes in solution. AB - The thermodynamics of electrochemical and complexation reactions involving the heterobimetallic triple-stranded helicates [MA(L5)3]n+ (M=Ru(II), Cr(III) and A=Ca(II), Lu(III)) reveal that solvation processes mask intramolecular intermetallic repulsions in solution, a phenomenon at the origin of the surprising stabilities of highly charged self-assembled polymetallic complexes in solution. A judicious combination of Born-Haber cycles and the Born equation restores the expected electrostatic trend in the gas phase, in which intermetallic interactions can be simply modeled using a standard Coulombic approach. Semiquantitative estimation and prediction of the contribution of the intermetallic repulsion to the total free energy of the formation of discrete polymetallic assemblies in solution become thus accessible. This point is crucial for programming stable metallosupramolecular architectures in solution. PMID- 17408263 TI - Organic-inorganic hybrids constructed of anderson-type polyoxoanions and oxalato bridged dinuclear copper complexes. AB - Two novel organic-inorganic hybrid compounds based on Anderson-type polyoxoanions, [Cu2(bpy)2(mu-ox)][Al(OH)7Mo6O17] (1) and [Cu2(bpy)2(mu ox)][Cr(OH)7Mo6O17] (2), have been synthesized and characterized by elemental analyses, IR, and X-ray powder diffraction. The crystal structures of 1 and 2 have been established by single-crystal X-ray diffraction, which reveals the presence of 1D chains constructed of alternating Anderson-type polyoxoanions and oxalato-bridged dinuclear copper complexes for both compounds and extensive hydrogen bonding that plays an important role in the formation of the 3D supramolecular network structures of 1 and 2. To elucidate the electronic properties and magnetic properties of the metal ions (Cu2+ or Cu2+ and Cr3+), EPR studies and magnetic susceptibility studies have been performed, respectively. The results are consistent with the structural feature of these compounds. PMID- 17408264 TI - Coexistence of five different copper(II)-phenanthroline species in the crystal packing of inorganic-metalorganic hybrids based on Keggin polyoxometalates and copper(II)-phenanthroline-oxalate complexes. AB - Reaction of in situ generated copper(II)-monosubstituted Keggin polyoxometalates and copper(II)-phenanthroline-oxalato complexes in ammonium or rubidium acetate buffers led to the formation of the hybrid inorganic-metalorganic compounds E4[Cu(phen)(H2O)4]2[Cu4(phen)4(H2O)4(ox)3]0.6[Cu2(phen)2(H2O)4(ox)]0.4[Cu(phen)(o )]0.8[{SiW11O39Cu(H2O)}2{Cu2(phen)2(ox)}].20H2O [E: Rb (1), NH4 (2)]. The two compounds have been characterized by means of elemental analysis, thermogravimetry, infrared and electron paramagnetic resonance spectroscopies, and magnetic susceptibility measurements, and their structures have been established by single-crystal X-ray diffraction. Both compounds are isostructural, and they contain a discrete bimolecular hybrid polyanion and several types of copper-phenanthroline complexes of variable nuclearity. The main structural features of these compounds are the presence of the new hybrid POM [{SiW11O39Cu(H2O)}2{Cu2(phen)2(mu-ox)}]10-, where the dinuclear copper-oxalato complex is sandwiched by two copper-monosubstituted POMs, and the coexistence of five different copper-phenanthroline species with nuclearities ranging from one to four. PMID- 17408265 TI - Synthesis and spectroscopic studies of cyclometalated Pt(II) complexes containing a functionalized cyclometalating ligand, 2-phenyl-6-(1H-pyrazol-3-yl)-pyridine. AB - Three new luminescent cyclometalated Pt(II) complexes, [Pt(L)Cl] (1), [Pt2(L-)2] (2), and [Pt(L)(PPh3)]ClO4 (3.ClO4) (where HL=2-phenyl-6-(1H-pyrazol-3-yl) pyridine), were synthesized and characterized by X-ray crystallography. HL represents a new class of C,N,Npyrazolyl cyclometalating ligands containing a Cphenyl, a Npyridyl, and a Npyrazolyl donor moiety, as well as a 1-pyrazolyl-NH, that can also be available for metal coordination and other chemical interactions. Complex 1 possesses intense intraligand transitions at 275-375 nm and moderately intense metal-to-ligand charge transfer (1MLCT) (dpi(Pt)-->pi*(L)) transition at 380-410 nm. The room temperature solid-state emission lambdamax of 1 occurs at 580 nm and is attributable to the 3MMLCT (dsigma*(Pt)-->pi*(L)) transition. It also displays strong phosphorescence in acetonitrile solutions at room temperature with an emission lambdamax at 514 nm, which can be tentatively assigned to the 3MLCT (pi*(L)-->dpi(Pt)) transition. Complex 1 can be deprotonated in organic solvents to yield a cycloplatinated dimer 2, which shows a relatively high room-temperature luminescent quantum yield of 0.59 in DMF (lambdamax=509 nm). Substitution of the ancillary chloro-ligand in 1 by triphenylphosphine yields 3, which also possesses a good room-temperature luminescent quantum yield of 0.52 in DMF (lambdamax=504 nm) and a better solubility in water. Complex 3 is synthesized to demonstrate the pH dependence of luminescent properties of this C,N,Npyrazolyl cyclometalated Pt(II) system. Such a pH response is ascribable to the protonation/deprotonation of the 1-pyrazolyl NH on the C,N,Npyrazolyl cyclometalating ligand. The pKa of the 1-pyrazolyl-NH in 3, measured in 1:2 (v/v) aqueous DMF solutions, is approximately 4.0. PMID- 17408266 TI - Single crystal 55Mn ENDOR of concanavalin a: detection of two Mn2+ sites with different 55Mn quadrupole tensors. AB - Concanavalin A is a member of the plant hemeagglutinin (or plant lectin) family that contains two metal binding sites; one, called S1, is occupied by Mn2+ and the other, S2, by Ca2+. 55Mn electron-nuclear double resonance (ENDOR) measurements were performed on a single crystal of concanavalin A at W-band (95 GHz, ~3.5 T) to determine the 55Mn nuclear quadrupole interaction in a protein binding site and its relation to structural parameters. Such measurements are easier at a high field because of the high sensitivity for size-limited samples and the reduction of second-order effects on the spectrum which simplifies spectral analysis. The analysis of the 55Mn ENDOR rotation patterns showed that two chemically inequivalent Mn2+ types are present at low temperatures, although the high-resolution X-ray structure reported only one site. Their quadrupole coupling constants, e2Qq/h, are significantly different; 10.7 +/- 0.6 MHz for Mand only -2.7 +/-0.6 MHz for M. The ENDOR data also refined the hyperfine coupling determined earlier by single-crystal EPR measurements, yielding a small but significant difference between the two: -262.5 MHz for M and -263.5 MHz for M. The principal z-axis for M is not aligned with any of the Mn-ligand directions, but is 25 off the Mn-asp10 direction, and its orientation is different than that of the zero-field splitting (ZFS) interaction. Because of the small quadrupole interaction of M the orientation dependence was very mild, leading to larger uncertainties in the asymmetry parameter. Nonetheless, there too z is not along the Mn-ligand bonds and is rotated 90 with respect to MnA. These results show, that similar to the ZFS, the quadrupolar interaction is highly sensitive to small differences in the coordination sphere of the Mn2+, and the resolution of the two types is in agreement with the earlier observation of a two-site conformational dynamic detected through the ZFS interaction, which is frozen out at low temperatures and averaged at room temperature. To account for the structural origin of the different e2Qq/h values, the electric field gradient tensor was calculated using the point-charge model. The calculations showed that a relatively small displacement of the oxygen ligand of asp10 can lead to differences on the order observed experimentally. PMID- 17408267 TI - Simultaneous measurement of orientational and spectral dynamics of single molecules in nanostructured host-guest materials. AB - Nanostructured host-guest materials are important for various applications in nanoscience, and therefore, a thorough understanding of the dynamics of the guest molecules within the host matrix is needed. To this aim we used single-molecule fluorescence techniques to simultaneously examine the spectral and the orientational behavior of single molecules in nanostructured porous host materials. Two types of host-guest systems have been investigated. First, oxazine 1 dye molecules were fixed rigidly in the channels of microporous AlPO4-5 crystals. Second, it was shown that terrylenediimide (TDI) dye molecules move in the mesoporous network of an uncalcined M41S thin film. In the first sample both spectral fluctuations ( approximately 5 nm) and rare spectral jumps (>10 nm) of the emission maximum were observed. However, the orientation of the emission dipole of the dye molecules remained constant. In contrast, the second system showed orientational dynamics as well as substantially more spectral dynamics. In this system the molecules were found to move between different regions in the host. The typical motion of the TDI molecules in the pores of M41S was not continuous but characterized by jumps between specific sites. Moreover, the spectral and orientational dynamics were correlated and arose directly from the different environments that were being explored by the mobile molecule. PMID- 17408268 TI - para-Hydrogen-induced polarization in heterogeneous hydrogenation reactions. AB - We demonstrate the creation and observation of para-hydrogen-induced polarization in heterogeneous hydrogenation reactions. Wilkinson's catalyst, RhCl(PPh3)3, supported on either modified silica gel or a polymer, is shown to hydrogenate styrene into ethylbenzene and to produce enhanced spin polarizations, observed through NMR, when the reaction was performed with H2 gas enriched in the para spin isomer. Furthermore, gaseous phase para-hydrogenation of propylene to propane with two catalysts, the Wilkinson's catalyst supported on modified silica gel and Rh(cod)(sulfos) (cod = cycloocta-1,5-diene; sulfos = O3S(C6H4)CH2C(CH2PPh2)3) supported on silica gel, demonstrates heterogeneous catalytic conversion resulting in large spin polarizations. These experiments serve as a direct verification of the mechanism of heterogeneous hydrogenation reactions involving immobilized metal complexes and can be potentially developed into a practical tool for producing catalyst-free fluids with highly polarized nuclear spins for a broad range of hyperpolarized NMR and MRI applications. PMID- 17408269 TI - Direct labeling of 5-methylcytosine and its applications. AB - Cytosine methylation is one of the most important epigenetic events, and much effort has been directed to develop a simple reaction for methylcytosine detection. In this paper, we describe the design of tag-attachable ligands for direct methylcytosine labeling and their application to fluorescent and electrochemical assays. The effect of the location of bipyridine substituents on the efficiency of osmium complexation at methylcytosine was initially investigated. As a result, a bipyridine derivative with a substituent at the C4 position showed efficient complexation at the methylcytosine residue of single stranded DNA in a reaction mixture containing potassium osmate and potassium hexacyanoferrate(III). On the basis of this result, a bipyridine derivative with a tag-attachable amino linker at the C4 position was synthesized. The efficiency of metal complex formation in the presence of the osmate and the synthetic ligand was clearly changed by the presence/absence of a methyl group at the C5 position of cytosine. The succinimidyl esters of functional labeling units were then attached to the bipyridine ligand fixed on the methylcytosine. These labels attached to methylcytosine enabled us to detect the target methylcytosine in DNA both fluorometrically and electrochemically. For example, we were able to fluorometrically obtain information on the methylation status at a specific site by means of fluorescence resonance energy transfer from a hybridized fluorescent DNA probe to a fluorescent label on methylcytosine. In addition, by the combination of electrochemically labeled methylcytosine and an electrode modified by probe DNAs, a methylcytosine-selective characteristic current signal was observed. This direct labeling of methylcytosine is a conceptually new methylation detection assay with many merits different from conventional assays. PMID- 17408270 TI - Membrane-grafted hyaluronan films: a well-defined model system of glycoconjugate cell coats. PMID- 17408271 TI - beta-Hydrogen Kinetic Effect. AB - A combined kinetic and DFT study of the uncatalyzed isomerization of cationic solvent complexes of the type cis-[Pt(R')(S)(PR3)2]+ (R' = linear and branched alkyls or aryls and S = solvents) to their trans isomers has shown that the reaction goes through the rate-determining dissociative loss of the weakly bonded molecule of the solvent and the interconversion of two geometrically distinct T shaped 14-electron three-coordinate intermediates. The Pt-S dissociation energy is strongly dependent on the coordinating properties of S and independent of the nature of R'. The energy barrier for the fluxional motion of [Pt(R')(PR3)2]+ is comparatively much lower ( approximately 8-21 kJ mol-1). The presence of beta hydrogens on the alkyl chain (R' = Et, Prn, and Bun) produces a great acceleration of the reaction rate. This accelerating effect has been defined as the beta-hydrogen kinetic effect, and it is a consequence of the stabilization of the transition state and of the cis-like three-coordinate [Pt(R')(PR3)2]+ intermediate through an incipient agostic interaction. The DFT optimization of [Pt(R')(PMe3)2]+ (R' = Et, Prn, and Bun) reproduces a classical dihapto Pt....eta2-HC agostic mode between the unsaturated metal and a dangling C-H bond. The value of the agostic stabilization energy (in the range of approximately 21 33 kJ mol-1) was estimated by both kinetic and computational data and resulted in being independent of the length of the hydrocarbon chain of the organic moiety. A better understanding of such interactions in elusive reaction intermediates is of primary importance in the control of reaction pathways, especially for alkane activation by metal complexes. PMID- 17408272 TI - A protein nanocarrier from charge-conversion polymer in response to endosomal pH. PMID- 17408273 TI - The acylation mechanism of CTX-M beta-lactamase at 0.88 a resolution. PMID- 17408274 TI - Highly diastereoselective synthesis of homoallylic alcohols bearing adjacent quaternary centers using substituted allylic zinc reagents. PMID- 17408275 TI - Preparation of 2-trialkylsiloxy- substituted 1,3-dienes and their diels alder/cross-coupling reactions. AB - [reaction: see text] 2-Triethylsiloxy-substituted 1,3-butadiene has been prepared in gram quantities from chloroprene via a simple synthetic procedure. Silatrane and catechol-substituted analogues of this main group element substituted diene were prepared by ligand exchange and characterized by X-ray crystallography in addition to standard spectroscopic techniques. Diels-Alder reactions of these dienes are reported as well as subsequent TBAF assisted/Pd-catalyzed Hiyama cross coupling reactions of those Diels-Alder adducts. PMID- 17408276 TI - Mechanism of rhodium-catalyzed carbene formation from diazo compounds. AB - [reaction: see text] A large and normal nitrogen-15 kinetic isotope effect of 1.035 +/- 0.003 provides direct support for the proposed mechanism for the rhodium-catalyzed carbene formation from diazo compounds, which involves the fast formation of a metal-diazo complex followed by rate-limiting extrusion of N2. The large magnitude of the KIE indicates extensive C-N bond fission in the transition state. PMID- 17408277 TI - Total synthesis of gymnorrhizol, an unprecedented 15-membered macrocyclic polydisulfide from the Chinese mangrove Bruguiera gymnorrhiza. AB - [reaction: see text] The total synthesis of gymnorrhizol, a naturally occurring macrocyclic polydisulfide with a new skeleton and a potent proteintyrosinephosphatase 1B inhibitor, was prepared in three steps, starting from (R)-1-bromo-3-chloroisopropanol and 1,3-dichloropropan-2-ol. PMID- 17408278 TI - Boronated enynes as versatile sources of stereodefined and skeletally diverse molecules. AB - [reaction: see text] The application of a one-pot palladium-catalyzed cycloisomerization of enynes 1/Diels-Alder cycloaddition/allylboration sequence efficiently generates tricyclic structures with complete control of the four stereogenic centers. Ruthenium and platinum catalysts perform distinct transformations providing other isomeric boron-substituted cyclic compounds. PMID- 17408279 TI - New three-component reaction: novel formation of a seven-membered ring by the unexpected reaction at the gamma-position of the beta-keto ester. AB - [reaction: see text] The novel three-component reaction of aromatic aldehydes, ethylenediamine, and beta-keto esters is described. In this reaction, beta-keto esters react at the gamma-position which is generally unreactive to produce the seven-membered ring compounds. Products have secondary amines and beta-enamino esters, which can serve in further functionalizations to produce molecular diversity. PMID- 17408280 TI - Diazo preparation via dehydrogenation of hydrazones with "activated" DMSO. AB - [reaction: see text] We report that "activated" dimethyl sulfoxide efficiently dehydrogenates hydrazones to the respective diazo species at -78 degrees C. Under optimized conditions, triethylamine hydrochloride is removed quantitatively by vacuum filtration to provide solutions of diazo compounds. Stable diazo species can be isolated in high yield, or alternatively, the direct treatment of these solutions with carboxylic acids provides esters. PMID- 17408281 TI - Function-oriented synthesis: studies aimed at the synthesis and mode of action of 1alpha-alkyldaphnane analogues. AB - [reaction: see text] An efficient synthetic route to the ABC tricyclic core of 1alpha-alkyldaphnanes has been developed. The conformational bias imparted by the C6-C9 oxo-bridge of BC-ring system 12 was used to elaborate the ABC-ring system precursor including the introduction of the beta-C5 hydroxyl group. A completely diastereoselective palladium-catalyzed enyne cyclization was then employed to establish the A-ring with a C1 appendage. PMID- 17408282 TI - An efficient oxidative lactonization of 1,4-diols catalyzed by Cp*Ru(PN) complexes. AB - [reaction: see text] An efficient oxidative lactonization of 1,4-diols in acetone is accomplished by the well-defined ruthenium catalyst, whose bifunctional nature underlies the high efficiency as well as unique chemo- and regioselectivity of the reaction which provides a rapid access to gamma-butyrolactones including flavor lactones hinokinin, and muricatacin. PMID- 17408283 TI - Synergistic effect between components of mixtures of cationic amphipaths in transfection of primary endothelial cells. AB - To date, the primary approach to improving the transfection properties of cationic lipids has been the synthesis of new kinds of cationic amphipaths. Recently, however, it was found that combining two cationic lipid derivatives having the same head group but tails of different chain lengths can provide another, and often superior, approach to higher transfection efficiency. For example, the combination of medium-chain and long-chain homologues of O ethylphosphatidylcholine transfected DNA into primary human umbilical artery endothelial cells (HUAECs) more than 30-fold more efficiently than did either compound separately. Here it is reported that this synergism of mixtures is not limited to O-ethylphosphatidylcholine homologues, but is also exhibited by other common cationic amphipathic transfection reagents; for example, combining DC-Chol (3beta-[N',N'-dimethylaminoethane)-carbamol] cholesterol), dimethylditetradecylammonium bromide, or DMTAP (1,2-dimyristoyl-3 trimethylammonium-propane) with EDOPC increased transfection significantly both in the absence and in the presence of serum. Furthermore, combining a poorer transfection agent-dimethyldioctadecylammonium bromide-with dimethylditetradecylammonium bromide increased transfection by about an order of magnitude with a maximum at an intermediate composition. Lack of synergy occurred with some mixtures, such as DMTAP and DOTAP (1,2-dioleoyl-3-trimethylammonium propane), in which case transfection activity was a linear function of composition both in the absence and presence of serum. Although the mechanism of enhanced transfection by mixtures is not fully understood, the existence of a number of optimal mixtures with diverse cationic compounds indicates that attention to mixture formulations can lead to greatly improved transfection by cationic amphipathic carriers. PMID- 17408284 TI - A new entry to polycyclic indole derivatives via intramolecular imino Diels-Alder reaction: observation of unexpected reaction. AB - A new, efficient, and highly diastereoselective synthesis of polycyclic indole derivatives through intramolecular imino Diels-Alder reaction of aminoanthraquinone with N-prenylated indole-2-carboxaldehydes in the presence of 20 mol % of triphenylphosphonium perchlorate (TPPP) is reported with extremely high cis selectivity in good yield. PMID- 17408285 TI - Ionic-liquid-promoted palladium-catalyzed multicomponent cyclocarbonylation of o iodoanilines and allenes to form methylene-2,3-dihydro-1H-quinolin-4-ones. AB - The palladium-catalyzed cyclocarbonylation reaction of o-iodoanilines with allenes and CO in 1-butyl-3-methylimidazolium hexafluorophosphate afforded 3 methylene-2,3-dihydro-1H-quinolin-4-ones in moderate to excellent yields under a low pressure (5 atm) of CO. The ionic liquid, as the solvent and promoter, enhances the efficiency of the cyclocarbonylation reaction. The recyclability of the system of ionic liquid/catalyst/ligand was also demonstrated. PMID- 17408286 TI - One-step synthesis of heterocyclic privileged medicinal scaffolds by a multicomponent reaction of malononitrile with aldehydes and thiols. AB - Heterocyclic privileged medicinal scaffolds involving pyridine, 1,4 dihydropyridine, chromeno[2,3-b]pyridine, and dihydro-1,4-dithiepine frameworks are prepared via a single-step multicomponent reaction of structurally diverse aldehydes with various thiols and malononitrile. Mechanistic studies of the synthetic pathway leading to pyridines reveal that 1,4-dihydropyridines undergo oxidation by the intermediate Knoevenagel adducts rather than by air oxygen. The use of o,o'-disubstituted aromatic aldehydes leads to the corresponding 1,4 dihydropyridines, whereas salicylic aldehydes result in chromeno[2,3-b]pyridines. Reactions of ethanedithiol as a thiol component produce dimeric pyridines with sterically unencumbered aldehydes, while o,o'-disubstituted aromatic aldehydes give dihydro-1,4-dithiepines. Thus, depending on the aldehyde and thiol types, diverse libraries of medicinally relevant compounds can be prepared by a simple one-step process involving no chromatography. PMID- 17408287 TI - Ozonolysis of unsaturated organotrifluoroborates. AB - Organotrifluoroborates are robust reagents capable of withstanding ozonolysis of remote alkenes, thus providing a new route to oxo-substituted organotrifluoroborates. The primary ozonides initially generated upon ozonolysis can be reduced with Zn/AcOH to afford the carbonyl compounds. Alternatively, capture of the carbonyl oxides with either an appropriate N-oxide or H2O easily gives the desired oxo-substituted organotrifluoroborates. Both unsaturated alkyltrifluoroborates and aryltrifluoroborates effectively participate in the reaction. The process provides oxo-functionalized organotrifluoroborates that cannot be prepared directly via either transmetalation or hydroboration protocols. PMID- 17408288 TI - Trisannelation of acrylates to 1,3,5-benzenetricarboxylates by a Pd(OAc)2/HPMoV/CeCl3/O2 system. AB - A new type of trisannelation reaction of acrylates through acetal formation was developed by Pd(OAc)2 combined with molybdovanadophosphoric acid (HPMo8V4) and Lewis acid under atmospheric dioxygen. Thus, the reaction of isobutyl acrylate in the presence of Pd(OAc)2, HPMo8V4, and CeCl3 under O2 (1 atm) in MeOH/AcOH afforded isobutyl 1,3,5-benzenetricarboxylate in fair yield. The reaction was found to proceed through the palladium-catalyzed acetalization of acrylate with methanol followed by the trisannelation reaction of the resulting acetal promoted by CeCl3. PMID- 17408289 TI - Intramolecular "hydroiminiumation and -amidiniumation" of alkenes: a convenient, flexible, and scalable route to cyclic iminium and imidazolinium salts. AB - Addition of a stoichiometric amount of HCl to alkenylaldimines, -formamidines, and -amidines results in the protonation of the sp2-nitrogen atom. The resulting alkenylaldiminium, -formamidinium, and -amidinium salts can be isolated and fully characterized, including single-crystal X-ray diffraction studies. Heating solutions of these salts induces ring closure cleanly and regioselectively via formal "exo" addition of the nitrogen-hydrogen bond to the pendent carbon-carbon double bond, affording the corresponding cyclic aldiminium, dihydroisoquinolinium, and imidazolinium salts. Of special interest, novel 4,4 disubstituted imidazolinium salts are accessible via this synthetic route. Similarly, addition of phosgene to alkenyl ureas and alkenyl amides, followed by gentle heating, cleanly affords C-chloro imidazolinium, and cyclic C-chloro iminium salts, respectively. Treatment of the latter with tetrakis(triphenylphosphine)palladium allows for the preparation of the first transition-metal complex bearing a cyclic arylaminocarbene as ligand. Deuterium labeling experiments suggest that the mechanism of the hydroiminiumation and amidiniumation reactions involves an intramolecular proton transfer to the double bond in the rate-determining step. This novel synthetic methodology gives access to a variety of N-heterocyclic carbene (NHC) and cyclic alkyl- and arylaminocarbene (CAAC) precursors. PMID- 17408290 TI - Hydrophilicity and the viscosity of interfacial water. AB - We measure the viscosity of nanometer-thick water films at the interface with an amorphous silica surface. We obtain viscosity values from three different measurements: friction force in a water meniscus formed between an oxide terminated W tip and the silica surface under ambient conditions; similar measurements for these interfaces under water; and the repulsive "drainage" force as the two surfaces approach at various speeds in water. In all three cases, we obtain effective viscosities that are approximately 10(6) times greater than that of bulk water for nanometer-scale interfacial separations. This enhanced viscosity is not observed when we degrade the hydrophilicity of the surface by terminating it with -H or -CH3. In view of recent results from other interfaces, we conclude that the criterion for the formation of a viscous interphase is the degree of hydrophilicity of the interfacial pair. PMID- 17408291 TI - Frictional drag and electrical manipulation of recombinant proteins in polymer supported membranes. AB - We establish a lipid monolayer supported by a polymer interface that offers advantages over conventional solid-supported membranes for determining the frictional drag at the membrane-protein interface as well as for electric field manipulation of membrane-anchored proteins. Polymer-supported monolayers with functional lipid anchors allow for the specific docking of His-tagged green fluorescent protein variants (His-EGFP and His-DsRed tetramer) onto the membrane surface at a defined surface density. In the first part, we measure the lateral diffusion coefficients of lipids and proteins and calculate the frictional drag at the protein-membrane interface. The second part deals with the electric field induced accumulation of recombinant proteins on a patterned surface. The mean drift velocity of proteins, which can be obtained analytically from the shape of the steady-state concentration gradient, can be controlled by tuning the interplay of electrophoresis and electroosmosis. The results demonstrate the potential of such molecular constructs for the local functionalization of solid substrates with membrane-associated proteins. PMID- 17408292 TI - A general route to prepare one- and three-dimensional carbon nanotube/metal nanoparticle composite nanostructures. AB - Adsorption of polyethyleneimine (PEI)-metal ion complexes onto the surfaces of carbon nanotubes (CNTs) and subsequent reduction of the metal ion leads to the fabrication of one-dimensional CNT/metal nanoparticle (CNT/M NP) heterogeneous nanostructures. Alternating adsorption of PEI-metal ion complexes and CNTs on substrates results in the formation of multilayered CNT films. After exposing the films to NaBH4, three-dimensional CNT composite films embedded with metal nanoparticles (NPs) are obtained. UV-visible spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy are used to characterize the film assembly. The resulting (CNT/M NP)n films inherit the properties from both the metal NPs and CNTs that exhibit unique performance in surface-enhanced Raman scattering (SERS) and electrocatalytic activities to the reduction of O2; as a result, they are more attractive compared to (CNT/polyelectrolyte)n and (NP/polyelectrolyte)n films because of their multifunctionality. PMID- 17408293 TI - Metal nanocrystals incorporated within pH-responsive microgel particles. AB - Cross-linked sterically stabilized latexes of approximately 250 nm diameter were synthesized by emulsion polymerization of 2-(diethylamino)ethyl methacrylate using a bifunctional oligo(propylene oxide)-based diacrylate cross-linker and a poly(ethylene oxide)-based macromonomer as the stabilizer at pH 9. These particles exhibit reversible swelling properties in water by adjusting the solution pH. At low pH, they exist as swollen microgels as a result of protonation of the tertiary amine units. Deswelling occurs above pH 7 [the effective pK(a) of poly(2-(diethylamino)ethyl methacrylate)], leading to the formation of the original compact latex particles. The swollen microgels can be used as nanoreactors: efficient impregnation with Pt nanoparticles can be achieved by incorporating precursor platinum compounds, followed by metal reduction. Dynamic light scattering was used to compare two methods of Pt nanoparticle impregnation with respect to the size and stability of the final Pt loaded microgel particles. In the first method, the H2PtCl6 precursor was added to hydrophobic latex particles at high pH, followed by metal reduction. In the second method, H2PtCl6 was added to hydrophilic swollen microgel particles at low pH, and then this metal salt was reduced in situ using NaBH4 and the pH was raised by the addition of base. Both the Pt salt-loaded (metalated) microgels and the final Pt nanoparticle-loaded microgels had well-defined structures that were independent of the synthesis route. Polymer-metal interactions were investigated by UV-visible absorption spectroscopy, which confirmed that the Pt salt was completely reduced to zero-valent Pt. Transmission electron microscopy and X-ray diffraction studies verified the formation of nanometer-sized Pt nanoparticles within these microgels, which can be used as recoverable colloidal catalyst supports for various organic reactions. PMID- 17408294 TI - Stability and state of aggregation of aqueous fibrinogen and dipalmitoylphosphatidylcholine lipid vesicles. AB - The stability and state of aggregation of aqueous fibrinogen (FB) and dipalmitoylphosphatidylcholine (DPPC) vesicles in water or buffer at 25 degrees C were studied with dynamic light scattering (DLS), UV-vis spectroturbidimetry (ST), and cryo-transmission electron microscopy (cryo-TEM). In water, when 1000 ppm (0.10 wt %) DPPC dispersions were prepared with a protocol including extensive sonication, they contained mostly vesicles and were quite clear, transparent, and stable for at least 30 days. FB mixtures with water (0.075 wt %) were quite unstable and biphasic. They formed large aggregates which eventually precipitated. The addition of DPPC vesicles into these unstable FB dispersions reversed FB aggregation and precipitation and produced stable translucent microdispersions. The inferred lipid/protein aggregates were limited in size, with average diameters ranging from 200 to 300 nm. In buffer, DPPC dispersions were also clear and quite stable, with average dispersed particles diameter of ca. 90 nm. FB dissolved in aqueous buffer and formed transparent and stable solutions. Adding salt to an aggregated FB dispersion in water reversed the aggregation. FB aggregated and redissolved in the presence of the citrate and after the citrate was removed. There was no effect of citrate (present in FB initially) in the FB aggregation or redissolution. FB molecules in buffer form dimers or higher aggregates. Their average aggregation number is 2, determined with Rayleigh scattering analysis of turbidity data. The average hydrodynamic diameter of FB solutions from DLS was 30 nm. Mixing a stable FB solution in buffer and a stable DPPC dispersion in buffer produced highly unstable mixtures, in which large aggregates precipitated. These results have implications in understanding the interactions of lipids and proteins in many biological applications and food processing applications. PMID- 17408295 TI - Formation and healing of micrometer-sized channel networks on highly mobile Au(111) surfaces. AB - Practical protocols are presented to reproducibly prepare micrometer-sized Au(111) substrates. Au(111) terraces of micrometer dimensions and atomic smoothness were crystallized by flame-annealing vacuum-deposited gold films on glass and on millimetric amorphous gold shots. Gold films and shots that were slowly cooled in a moderately applied stream of nitrogen gas exhibited large and stable crystal surfaces with Au(111) morphologies. Similarly, flame-annealed gold samples cooled with another protocol--in much rougher streams of nitrogen gas- produced morphologically unstable and highly mobile Au(111) layers. Within the first hour after preparation, however, rapid microscale restructuring in the layers produced complex morphologies of hexagonal channel networks and islands that were predominantly triangular. These channeled gold layers fused slowly in the following hours, with velocities of 0.01-0.2 A/s, as quantified by digital image correlation (DIC). Atomically smooth, stable, and predominantly triangular Au(111) terraces on the scale of micrometers were observed approximately 24 h after the sample preparations. PMID- 17408296 TI - Particle assembly on surface features (patterned surfaces). AB - Irreversible adsorption (deposition) of spherical particles on surface features of various shapes (collectors) was studied using the random sequential adsorption (RSA) model. The collectors in the form of linear line segments, semicircles, and circles were considered. Numerical simulation of the Monte Carlo type enabled one to determine particle configurations, the jamming coverage, and the end to end length of particle monolayers for various collector length (L) to particle size (d) ratio L = L/d. It was revealed that the jamming coverage for linear collectors Theta'(infinity) increases for L > 2 according to a linear dependence with respect to 1/L. For 2 > L > 1, a parabolic dependence of Theta'(infinity) on 1/L was predicted, characterized by the maximum value of Theta'(infinity) = 1.125 for L = 4/3. These dependencies allowed one to formulate an equation determining the length of nanostructures on surfaces if the averaged number of adsorbed particles is known. It was also predicted that the end to end length of the monolayer on a linear collector /L increased linearly with 1/L for L > 2. For 2 > L > 1 the dependence of /L on L was approximated by a polynomial expression, exhibiting a maximum of /L = 1.17 for L = 1.45. In the case of circular collectors, the jamming coverage was found to be substantially smaller for the same value of 1/L. It was demonstrated that the theoretical results are in agreement with our preliminary experimental data obtained for latex particles adsorbing on polyelectrolyte modified mica and on patterned surfaces obtained by a polymer-on-polymer stamping technique of gold covered silicon (Zheng et al. Langmuir 2002, 18, 4505). PMID- 17408297 TI - Amphiphilic perylenetretracarboxyl diimide dimer and its application in field effect transistor. AB - A novel perylene diimide (PDI) derivative with typical amphiphilic character, 2, was designed and prepared. The spectroscopic studies on this compound in solution revealed the face-to-face dimeric configuration and effective pi-pi interaction between the two perylene rings. This amphiphilic PDI derivative was fabricated into highly ordered films by Langmuir-Blodgett (LB) technique and fabricated into an organic field effect transistor (OFET), which shows carrier mobility around 0.05 cm(2) V(-1) s(-1) and current modulation of 10(3). This OFET performance is much better than that of monomeric PDI 1 and can be attributed to the unique face to-face structure of 2, which promotes the interactions between neighboring PDI ring in LB film as indicated by the pi-A isotherms and UV-vis absorption. PMID- 17408298 TI - Time lapse confocal microscopy studies of bacterial adhesion to self-assembled monolayers and confirmation of a novel approach to the thermodynamic model. AB - In this study, we use thermodynamic theory to develop a novel model that allows for the quantitative determination of the Gibbs free energy of adhesion for the initial bacterial attachment process. This model eliminates the need to calculate interfacial free energies and instead relies on easily measurable contact angles to determine DeltaG(adh). We experimentally verify our model using real-time observation of the initial attachment of Pseudomonas putida to methyl- and hydroxyl-terminated self-assembled monolayers. We also test the applicability of our model to a variety of experimental conditions using data available in the literature. We show that the initial attachment process is governed by dispersion forces and is accurately predicted by our model. Also, we find that our model is simple to apply and accurate for a variety of experimental conditions. PMID- 17408299 TI - Improving hydrophilicity and protein resistance of poly(vinylidene fluoride) membranes by blending with amphiphilic hyperbranched-star polymer. AB - To endow hydrophobic poly(vinylidene fluoride) (PVDF) membranes with reliable hydrophilicity and protein resistance, an amphiphilic hyperbranched-star polymer (HPE-g-MPEG) with about 12 hydrophilic arms in each molecule was synthesized by grafting methoxy poly(ethylene glycol) (MPEG) to the hyperbranched polyester (HPE) molecule using terephthaloyl chloride (TPC) as the coupling agent and blended with PVDF to fabricate porous membranes via phase inversion process. The chemical composition changes of the membrane surface were confirmed by X-ray photoelectron spectroscopy (XPS), and the membrane morphologies were measured by scanning electron microscopy (SEM). Water contact angle, static protein adsorption, and filtration experiments were used to evaluate the hydrophilicity and anti-fouling properties of the membranes. It was found that MPEG segments of HPE-g-MPEG enriched at the membrane surface substantially, while the water contact angle decreased as low as 49 degrees for the membrane with a HPE-g MPEG/PVDF ratio of 3/10. More importantly, the water contact angle of the blend membrane changed little after being leached continuously in water at 60 degrees C for 30 days, indicating a quite stable presence of HPE-g-MPEG in the blend membranes. Furthermore, the blend membranes showed lower static protein adsorption, higher water and protein solution fluxes, and better water flux recovery after cleaning than the pure PVDF membrane. PMID- 17408300 TI - Spontaneous formation of micrometer-size inorganic peapods. AB - We show that polyoxometalate (ammonium phosphomolybdate) Keggin in aqueous dispersions upon sonication spontaneously transforms into micrometer-sized, peapod-shaped structures. The formation of these peapods is preceded by the generation of spherical aggregates. The particles have been characterized experimentally by time-resolved dynamic light scattering (DLS), transmission electron microscopy (TEM), and scanning TEM with a high-angle annular dark field detector (STEM-HAADF) for energy-dispersive X-ray (STEM/EDX) elemental analyses. A pathway for the phenomenon is proposed. PMID- 17408301 TI - Nanocrystalline nanowires: III. Electrons. AB - Nanocrystalline nanowires (NCNW) are fragments of bulk crystals that are infinite in only one direction and typically have some rotational symmetry around this direction. Electron eigenstates belonging to the symmetry labels (k,m) (wavevector and rotational quantum number) are discussed. The rotational quantum number simplifies discussion of optical properties. For m not equal 0, the +/-m degeneracy allows orbital magnetism. The simplest sensible model which is more complex than a one-dimensional chain is solved. Methods are suggested for incorporating rotational symmetry into preexisting codes with three-dimensional translations. PMID- 17408302 TI - "Quantum coaxial cables" for solar energy harvesting. AB - Type II core-shell nanowires based on III-V and II-VI semiconductors are designed to provide the highly desirable but not readily available feature--efficient charge separation--and concurrently address the different material challenges specific for a few key renewable energy applications: including hydrogen generation via photoelectrochemical water splitting, dye-sensitized solar cells, and conventional solar cells. They also open up new avenues for studying novel physics and material sciences in reduced dimensionality of very unusual quasi-one dimensional systems. A first-principles density function theory within the local density approximation (LDA) is used for the electronic structure calculation and a valence-force-field method for the structural relaxation, and empirical corrections to the LDA errors are applied. PMID- 17408303 TI - Variables influencing interactions of untargeted quantum dot nanoparticles with skin cells and identification of biochemical modulators. AB - Skin cells (NHEK) take up untargeted quantum dots (QD) with surface polyethylene glycol (PEG), amines, and carboxylic acids, but the mechanisms are unknown. Time courses of QD-NHEK interactions were determined and effects of QD surface coating, temperature, culture medium supplements and inhibitors of the cell cycle and endocytosis identified. The magnitude of QD-NHEK interactions was coating dependent. Low-temperature or unsupplemented medium decreased QD-NHEK interactions. Biochemical inhibitors were identified that attenuate and potentiate QD-NHEK interactions. These results are important for understanding and controlling interactions of untargeted QD with cells. PMID- 17408305 TI - Drug-induced liver injury. AB - Drug-induced liver injury is a frequent cause of hepatic dysfunction. Reliably establishing whether the liver disease was caused by a drug requires the exclusion of other plausible causes and the search for a clinical drug signature. The drug signature consists of the pattern of liver test abnormality, the duration of latency to symptomatic presentation, the presence or absence of immune-mediated hypersensitivity and the response to drug withdrawal. Determination of causality also includes an evaluation of individual susceptibility to drug-induced liver injury. This susceptibility is governed by both genetic and environmental factors. Components of the drug signature in conjunction with certain risk factors have been incorporated into formal scoring systems that are predictive of the likelihood of drug-induced liver injury. The most validated scoring system is the Roussel-Uclaf causality assessment method, which nonetheless retains certain imperfections. Mitigating the potential for drug-induced liver injury is achieved by the identification of toxicity signals during clinical trials and the monitoring of liver tests in clinical practice. There are three signals of liver toxicity in clinical trials: (i) a statistically significant doubling (or more) in the incidence of serum alanine aminotransferase (ALT) elevation >3 x the upper limit of normal (ULN); (ii) any incidence of serum ALT elevation >8-10 x ULN; and (iii) any incidence of serum ALT elevation >3 x ULN accompanied by a serum bilirubin elevation >2 x ULN. Monitoring of liver tests in clinical practice has shown unconvincing efficacy, but where a benefit risk analysis would favour continued therapy, monthly monitoring may have some benefit compared with no monitoring at all. With rare exception, treatment of drug-induced liver injury is principally supportive. Drug toxicity is the most common cause of acute liver failure, defined as a prolonged prothrombin time (international normalised ratio > or =1.5) and any degree of mental alteration occurring <26 weeks after the onset of illness in a patient without pre-existing cirrhosis. A patient who meets these criteria must be evaluated for liver transplantation. The pathogenesis of drug-induced liver injury can be examined on the basis of the two principal patterns of injury. The hepatocellular pattern is characterised by a predominant rise in the level of transaminases and results from the demise of hepatocytes by means of either apoptosis or necrosis. The cholestatic pattern is characterised by a predominant rise of the serum alkaline phosphatase level and usually results from injury to the bile ductular cells either directly by the drug or its metabolite, or indirectly by an adaptive immune response. PMID- 17408306 TI - Systematic review of piperacillin-induced neutropenia. AB - Because penicillin agents are implicated in granulopoiesis inhibition, healthcare professionals frequently consider discontinuation of such therapy in patients with decreasing white blood cell counts. No systematic review to date has described piperacillin and the patient population at risk for this adverse drug reaction (ADR). This review sought to assess the occurrence of piperacillin induced neutropenia, describe characteristics of affected patients and assess the reporting modalities that most accurately classify this ADR. Case reports, cohort studies and clinical trials identified by comprehensive searches of PubMed and the US FDA Adverse Event Reporting System (AERS) database were reviewed for patient demographics, duration and dose of piperacillin or piperacillin tazobactam treatment and the occurrence of neutropenia. Causality assessments were performed. Six published case reports, three cohort studies, 178 clinical trials and two compilations of phase I-III trials were reviewed. Review of case reports was notable in that the duration of beta-lactam therapy prior to the noting of leukopenia always exceeded 15 days. No deaths were recorded in this group. Among 13,816 patients enrolled in non-neutropenic fever studies, the occurrence of piperacillin-induced neutropenia was rare: five patients (0.04%) developed neutropenia; none died. The demographics for this group were poorly documented. Through the AERS database, we identified 366 unique cases of piperacillin or piperacillin-tazobactam-induced haematological abnormalities, including neutropenia (n = 183, 50.0%), leukopenia, (n = 99, 27%), agranulocytosis (n = 58, 15.8%) and others. In 62 cases, patients received between 1 and 14 days of therapy (mean 7.7 + 4.1 days). Overall, there were 82 (22.4%) deaths. Reports of haematological ADRs among patients receiving piperacillin or piperacillin-tazobactam are rare. Report of neutropenia associated with piperacillin usage prior to 15 days of therapy is a novel finding that requires further evaluation. Current reporting methods poorly characterise patient groups at risk. PMID- 17408307 TI - Postmarketing analysis of medicines: methodology and value of the spanish case control study and surveillance system in preventing birth defects. AB - There are many surveillance systems of congenital defects all over the world; several of them have developed specific approaches to generate and test selected hypotheses regarding human teratogens. However, to the best of our knowledge, none of them have a permanent and systematised programme for the study of the risk and safety of drugs. The aim of this article is to describe the research programme on the potential effects of drugs in pregnancy followed by the Spanish Collaborative Study of Congenital Malformations (ECEMC), which is a permanent ongoing case-control study and surveillance system. The programme to analyse drugs includes a continuous and systematic study on the potential effects of medicines used during pregnancy. This programme has several characteristics that make it different from other current systems: (i) the collection of numerous datapoints (up to 312 per infant) in a case-control design; (ii) the use of a versatile and specific coding of birth defects; (iii) a specific programme for the continuous analysis of the potential effects of each type of drugs used during pregnancy that has been developed specifically for the ECEMC methodology, including its dysmorphological coding system. The description of the ECEMC's approach to surveillance of the effects of drug use during pregnancy may help researches in this area, particularly those using data from birth defects registries. PMID- 17408308 TI - Mometasone furoate nasal spray: a review of safety and systemic effects. AB - The development of corticosteroids that are delivered directly to the nasal mucosa has alleviated much of the concern about the systemic adverse effects associated with oral corticosteroid therapy. However, given the high potency of these drugs and their widespread use in the treatment of allergic rhinitis, it is important to ensure that intranasal corticosteroids have a favourable benefit risk ratio. One agent that typifies the systemic safety found in the majority of intranasal corticosteroids is mometasone furoate nasal spray, a potent and effective treatment for seasonal and perennial allergic rhinitis and nasal polyposis. Mometasone furoate does not reach high systemic concentrations or cause clinically significant adverse effects. Results from pharmacokinetic studies in adults and children suggest that systemic exposure to mometasone furoate after intranasal administration is negligible. This is probably because of the inherently low aqueous solubility of mometasone furoate, which allows only a small fraction of the drug to cross the nasal mucosa and enter the bloodstream, and because a large amount of the administered drug is swallowed and undergoes extensive first-pass metabolism. There is no clinical evidence that mometasone furoate nasal spray suppresses the function of the hypothalamus-pituitary-adrenal axis when the drug is administered at clinically relevant doses (100-200 microg/day); consequently, mometasone furoate nasal spray has not been associated with growth inhibition in children. The safety and tolerability of mometasone furoate nasal spray have been rigorously assessed in clinical trials involving approximately 4,500 patients, with epistaxis, headache and pharyngitis being the most common adverse effects associated with treatment in adolescents and adults. The clinical effectiveness of mometasone furoate nasal spray, coupled with its agreeable safety and tolerability profile, confirms its favourable benefit-risk ratio. PMID- 17408309 TI - Drug-induced yawning: a review of the French pharmacovigilance database. AB - OBJECTIVE: To review the reports with 'yawning' as an adverse drug reaction (ADR) reported into the French Pharmacovigilance Database. METHODS: All the observations with 'yawning' reported in the French Pharmacovigilance Database until December 2004 were reviewed. We recorded drug(s) involved, characteristics of patients (age, sex and underlying disease) and of ADR (seriousness, delay in occurrence, evolution, imputability). RESULTS: Twenty-eight reports were recorded between 1985 and December 2004. The sex ratio of the patients included in these reports was 1.5 and the mean age was 46.2 (2-78) years. Thirty-eight drugs were involved, mainly serotoninergic agents (serotonin reuptake inhibitors [12]), dopaminergic agents (levodopa [3], dopamine agonists [3], monoamine oxidase B inhibitor [1]), opioids (morphine [1], methadone [1], buprenorphine [1], dextromethorphan [1]), benzodiazepines (4) and sodium channel inhibitors (lidocaine [2], flecainide [1]). Four ADRs were rated 'serious' (leading to hospitalisation). Patient outcome was usually favourable after drug withdrawal. CONCLUSION: Despite its necessary methodological drawbacks (mainly under reporting), this study reveals that several drugs may induce yawning in humans. Our work also indicates that stimulation of central dopamine or serotonin receptors elicits yawning in humans. This study underlines the role of several drugs in yawning and shows that this ADR is not systematically listed in the summary product characteristic even when it can be explained by the pharmacodynamic properties of the drugs. PMID- 17408310 TI - Application of the bradford hill criteria to assess the causality of cisapride induced arrhythmia: a model for assessing causal association in pharmacovigilance. AB - INTRODUCTION: The Bradford Hill criteria are a widely used, useful tool for the assessment of biomedical causation. We have examined their application to pharmacovigilance using the example of cisapride-induced QTc interval prolongation/arrhythmia. METHODS: A literature search was conducted using MEDLINE, EMBASE, Reactions Weekly and regulatory websites to identify evidence for the association between cisapride and QTc interval prolongation/arrhythmia that had been published in the English language. Two hundred and five publications were identified as being potentially suitable for the study. After excluding irrelevant articles, studies on high-risk populations and review articles, 70 publications were assessed using the Bradford Hill criteria. These included 24 case reports, case series or spontaneous report summaries; eight epidemiological studies; 22 clinical studies; and 16 experimental (in vivo and in vitro) publications. RESULTS: The most compelling evidence for an association between cisapride use and QTc interval prolongation/arrhythmia came from case/spontaneous reports and biological plausibility. Considering the rare incidence of serious cardiac events, these criteria formed the basis for the strength of the association. The number of reports from different populations showed consistency. Specificity was supported by clinical and cardiographic characterisation of the events. There were temporal relationships between the events and the initiation of cisapride treatment, increases in the dosage and the receipt of interacting medications. The relationships between the adverse events and the latter two factors exhibited biological gradients. Experimental evidence could be found from biological models, as well as reports of positive dechallenge and/or rechallenge found in individual patients. Cisapride was found to bind the human ether-a-go-go-related gene (HERG) potassium channel, which provides a plausible mechanism for QTc interval prolongation/arrhythmia. Other QTc interval prolonging/arrhythmic drugs that also bind to HERG provided an analogy for cisapride causing QTc interval prolongation/arrhythmia via this mechanism. The evidence provided by clinical studies was inconsistent, and epidemiological studies failed to demonstrate an association. Nevertheless, this did not prevent the assessment of causation. DISCUSSION: This study showed how different types of evidence found in pharmacovigilance can be evaluated using the Bradford Hill criteria. Further work is required to examine how the criteria can be applied to different types of adverse events and how they may be applied to pharmacovigilance. PMID- 17408311 TI - Safety monitoring of a new pentavalent vaccine in the expanded programme on immunisation in Ghana. AB - BACKGROUND AND OBJECTIVE: Safety monitoring of vaccines used in expanded programmes on immunisation is important in all countries, including those with limited resources. As the rates of target diseases decrease, parents become less accepting of even minor common adverse events. Identification, detection, prevention and appropriate communication of adverse events following immunisation (AEFI) are therefore essential to preserve the integrity of immunisation programmes and protect public health. The objective of this study was to document the occurrence of common minor AEFI associated with a newly introduced pentavalent vaccine for routine immunisation in Ghana's expanded programme on immunisation. METHODS: A prospective descriptive study on AEFI associated with the administration of a pentavalent diphtheria, tetanus, pertussis, hepatitis B and Haemophilus influenzae type B (DTP-hepatitis B vaccine/Hib vaccine) vaccine that is part of the Expanded Programme on Immunisation was carried out in four locations in Accra, Ghana. These locations were the nation's premier teaching hospital (the Korle-Bu Teaching Hospital) two urban polyclinics (the Mamprobi and Ussher Town polyclinics) and a community immunisation centre (the Zongo Junction Immunisation Centre).A total of 406 infants were recruited for the study. Upon receipt of signed informed consent from the parents/guardians of the infants, the parents/guardians were supplied with a pink card that functioned as a pseudo diary for recording AEFI that occurred at home and for measuring and noting the sizes of any injection-site swellings that might occur. It also enabled each participant to obtain free medical care at the Department of Child Health, Korle Bu Teaching Hospital for the duration of the study (from September 2003 to December 2004) and until the child was 12 months old. Information about the occurrence of AEFI was actively solicited during each visit for immunisation and also at a visit 4 weeks after administration of the last dose of pentavalent vaccine, when participants were asked to report to the respective immunisation centres for the specific purpose of reporting any AEFI which might have occurred in the intervening period. These AEFI were analysed separately from those reported to the dedicated hospital unit at the Department of Child Health, Korle Bu Teaching Hospital, since the AEFI reported to that unit were all verified and recorded by trained physicians. RESULTS: Of the 406 infants, 368 completed the study, whereas 38 defaulted or were lost to follow-up. There were 104 attendances to report cases of suspected AEFI requiring physician attention at the Department of Child Health, Korle-Bu Teaching Hospital. These attendances were made by 74 patients who reported 190 events; notable among these were cough (26.3% of all AEFI reported to the hospital), fever (17.4%), common cold (12.1%), vomiting (7.4%) and diarrhoea (6.8%). Three of these visits involved AEFI that were classified as 'serious', since they required hospitalisation, but all three were considered to be unlikely to be related to vaccine administration. In addition, actively solicited information on AEFI following immunisation from 921 individual interviews with the parents/guardians of immunised infants during the follow-up visits resulted in reports of 259 events being reported, the most common, according to crude incidence rates, being fever (14.7%), common cold (3.8%), crying (3%) and cough (2.8%). CONCLUSION: The results of this study show agreement with safety studies on vaccines containing identical or similar antigens performed elsewhere and indicate the safety and tolerability of the pentavalent DTP-hepatitis B vaccine/Hib vaccine in Ghanaian children. PMID- 17408313 TI - Medication nonadherence: a national epidemic among America's seniors. PMID- 17408312 TI - The influence of primary care prescribing rates for new drugs on spontaneous reporting of adverse drug reactions. AB - INTRODUCTION: Adverse drug reaction (ADR) reporting makes a vital contribution to pharmacovigilance, although the factors that influence the reporting rate remain unclear. The aim of this study was to investigate whether the variation in the rate of reporting of suspected ADRs in different regions of Scotland was explained by differences in local prescribing practice and to quantify the extent of this influence. METHODS: Population and primary care prescribing data were obtained for ten geographical areas based on the 15 administrative regions of the National Health Service in Scotland. All reports of suspected ADRs received from within Scotland for 2000 and 2001 were available from the regional monitoring centre (Committee on Safety of Medicines, Scotland). The primary analysis was based on 14 medications that appeared in the 'top ten' list for the frequency of reported ADRs for either year. Reporting rates for each area were expressed both in terms of population (reports per million people) and in terms of estimated exposure to those medications in primary care (reports per 1000 prescriptions). For each analysis, the Pearson correlation coefficient between reporting and prescribing data was calculated using SPSS software. RESULTS: The 'top ten' medications accounted for 1715 of 2817 (60.9%, 95% CI 59.1, 62.7) ADR reports but only 2.2 million out of a total of 128 million primary care prescriptions (1.7%). Although there was a 3-fold geographical variation in the per-population ADR reporting rate, there was a close correlation between local reporting of ADRs and prescribing of the index medications (p = 0.66, p = 0.04, respectively). This implies that 44% of the observed variation in reporting rate can be attributed to variation in prescribing within the same population. DISCUSSION: Spontaneous ADR reporting in Scotland over the 2 years studied was highly concentrated on a small number of medications that were under intensive surveillance. Although there was a 3-fold variation in reporting rates from individual geographic areas when corrected for the size of the population, primary care prescribing data showed nearly half of this local variation in reporting rates could be explained by differences in prescribing. This study highlights the importance of considering prescribing practice when interpreting spontaneous ADR reporting data. PMID- 17408314 TI - Antiplatelet therapy in the treatment of atherothrombotic disease: considering the evidence. AB - Aspirin should be used to treat patients with acute myocardial infarction (MI) and continued indefinitely to reduce vascular death, nonfatal MI, and nonfatal stroke. Clopidogrel added to aspirin is beneficial in the treatment of patients with acute ST-elevation MI. Patients with unstable angina or non-ST-elevation MI should be treated with aspirin plus clopidogrel for at least 9 months to reduce the risk vascular death, nonfatal MI, and nonfatal stroke. Patients with prior MI should be treated indefinitely with aspirin and with clopidogrel if aspirin is contraindicated. Patients with ischemic stroke should be treated with either aspirin or clopidogrel indefinitely. Extended-release dipyridamole plus low-dose aspirin is more efficacious than low-dose aspirin but is associated with an insignificant increase in nonfatal MI and vascular death than low-dose aspirin. Clopidogrel is more effective than aspirin in reducing the risk of vascular death, nonfatal MI, and nonfatal stroke in patients with peripheral arterial disease. PMID- 17408315 TI - Early identification of cognitive deficits: preclinical Alzheimer's disease and mild cognitive impairment. AB - With the projected dramatic increase in the number of people who will be diagnosed with Alzheimer's disease (AD) in the coming years, interest is growing in identifying and treating adults at high risk for developing the disorder. Recent research suggests that individuals who will go on to receive a diagnosis of AD exhibit deficits in cognitive performance years beforehand. Those with mild cognitive impairment (MCI), for example, have characteristic cognitive deficits, such as memory loss, and convert to a diagnosis of AD at a faster rate than cognitively healthy controls. MCI has thus become a focus of research because it may help identify high-risk individuals for whom prophylactic treatments designed to slow the progress toward AD can be prescribed. After describing the diagnostic criteria and dementia outcomes associated with MCI, this article discusses several challenges to the study of cognitive impairment before the diagnosis of AD. PMID- 17408316 TI - Asymptomatic pigmented lesion on left ear. PMID- 17408317 TI - A biomechanical study of periacetabular defects and cement filling. AB - Periacetabular bone metastases cause severe pain and functional disability in cancer patients. Percutaneous acetabuloplasty (PCA) is a minimally invasive, image-guided procedure whereby cement is injected into lesion sites. Pain relief and functional restoration have been observed clinically; however, neither the biomechanical consequences of the lesions nor the effectiveness of the PCA technique are well understood. The objective of this study was to investigate how periacetabular lesion size, cortex involvement, and cement modulus affect pelvic bone stresses and strains under single-legged stance loading. Experiments were performed on a male cadaver pelvis under conditions of intact, periacetabular defect, and cement-filling with surface strains recorded at three strain gage locations. The experimental data were then employed to validate three-dimensional finite element models of the same pelvis, developed using computed tomography data. The models demonstrated that increases in cortical stresses were highest along the posterior column of the acetabulum, adjacent to the defect. Cortical stresses were more profoundly affected in the presence of transcortical defects, as compared to those involving only trabecular bone. Cement filling with a modulus of 2.2 GPa was shown to restore cortical stresses to near intact values, while a decrease in cement modulus due to inclusion of BaSO(4) reduced the restorative effect. Peak acetabular contact pressures increased less than 15% for all simulated defect conditions; however, the contact stresses were reduced to levels below intact in the presence of either cement filling. These results suggest that periacetabular defects may increase the vulnerability of the pelvis to fracture depending on size and cortical involvement and that PCA filling may lower the risk of periacetabular fractures. PMID- 17408318 TI - Deterministic material-based averaging theory model of collagen gel micromechanics. AB - Mechanics of collagen gels, like that of many tissues, is governed by events occurring on a length scale much smaller than the functional scale of the material. To deal with the challenge of incorporating deterministic micromechanics into a continuous macroscopic model, we have developed an averaging-theory-based modeling framework for collagen gels. The averaging volume, which is constructed around each integration point in a macroscopic finite-element model, is assumed to experience boundary deformations homogeneous with the macroscopic deformation field, and a micromechanical problem is solved to determine the average stress at the integration point. A two-dimensional version was implemented with the microstructure modeled as a network of nonlinear springs, and 500 segments were found to be sufficient to achieve statistical homogeneity. The method was then used to simulate the experiments of Tower et al. (Ann. Biomed. Eng., 30, pp. 1221-1233) who performed uniaxial extension of prealigned collagen gels. The simulation captured many qualitative features of the experiments, including a toe region and the realignment of the fibril network during extension. Finally, the method was applied to an idealized wound model based on the characterization measurements of Bowes et al. (Wound Repair Regen., 7, pp. 179-186). The model consisted of a strongly aligned "wound" region surrounded by a less strongly aligned "healthy" region. The alignment of the fibrils in the wound region led to reduced axial strains, and the alignment of the fibrils in the healthy region, combined with the greater effective stiffness of the wound region, caused rotation of the wound region during uniaxial stretch. Although the microscopic model in this study was relatively crude, the multiscale framework is general and could be employed in conjunction with any microstructural model. PMID- 17408319 TI - Effects of initial-fixed charge density on pH-sensitive hydrogels subjected to coupled pH and electric field stimuli: a meshless analysis. AB - In this paper, we study the effects of initial fixed-charge density on the response behavior of pH-sensitive hydrogels subjected to coupled stimuli, namely, solution pH and externally applied electric field. This is the first instance in which a coupled stimuli numerical analysis has been carried out for these polymer gels, which are used as active sensing/actuating elements in advanced biomicroelectromechanical systems devices. In this work, a chemo-electro mechanical formulation, termed the multi-effect-coupling pH-stimulus (MECpH) model, is first presented. This mathematical model takes into account the ionic species diffusion, electric potential coupling, and large mechanical deformation. In addition, a correlation between the diffusive hydrogen ions and fixed-charge groups on the hydrogel polymeric chains is established based on the Langmuir absorption isotherm, and incorporated accordingly into the MECpH model. To solve the resulting highly nonlinear and highly coupled partial differential equations of this mathematical model, the Hermite-Cloud method, a novel true meshless technique, is employed. To demonstrate the accuracy and robustness the MECpH model, computed numerical results are compared with experimental data available from literature. Following this validation, several numerical studies are carried out to investigate the effects of initial fixed-charge density on the volumetric variations of these pH-stimulus-responsive hydrogels when immersed in buffered solutions. PMID- 17408320 TI - A biphasic model for micro-indentation of a hydrogel-based contact lens. AB - The stiffness and hydraulic permeability of soft contact lenses may influence its clinical performance, e.g., on-eye movement, fitting, and wettability, and may be related to the occurrence of complications; e.g., lesions. It is therefore important to determine these properties in the design of comfortable contact lenses. Micro-indentation provides a nondestructive means of measuring mechanical properties of soft, hydrated contact lenses. However, certain geometrical and material considerations must be taken into account when analyzing output force displacement (F-D) data. Rather than solely having a solid response, mechanical behavior of hydrogel contact lenses can be described as the coupled interaction between fluid transport through pores and solid matrix deformation. In addition, indentation of thin membranes ( approximately 100 microm) requires special consideration of boundary conditions at lens surfaces and at the indenter contact region. In this study, a biphasic finite element model was developed to simulate the micro-indentation of a hydrogel contact lens. The model accounts for a curved, thin hydrogel membrane supported on an impermeable mold. A time-varying boundary condition was implemented to model the contact interface between the impermeable spherical indenter and the lens. Parametric studies varying the indentation velocities and hydraulic permeability show F-D curves have a sensitive region outside of which the force response reaches asymptotic limits governed by either the solid matrix (slow indentation velocity, large permeability) or the fluid transport (high indentation velocity, low permeability). Using these results, biphasic properties (Young's modulus and hydraulic permeability) were estimated by fitting model results to F-D curves obtained at multiple indentation velocities (1.2 and 20 microm/s). Fitting to micro-indentation tests of Etafilcon A resulted in an estimated permeability range of 1.0 x 10(-15) to 5.0 x 10(-15) m(4)N s and Young's modulus range of 130 to 170 kPa. PMID- 17408321 TI - One-dimensional models of the human biliary system. AB - This paper studies two one-dimensional models to estimate the pressure drop in the normal human biliary system for Reynolds number up to 20. Excessive pressure drop during bile emptying and refilling may result in incomplete bile emptying, leading to stasis and subsequent formation of gallbladder stones. The models were developed following the group's previous work on the cystic duct using numerical simulations. Using these models, the effects of the biliary system geometry, elastic property of the cystic duct, and bile viscosity on the pressure drop can be studied more efficiently than with full numerical approaches. It was found that the maximum pressure drop occurs during bile emptying immediately after a meal, and is greatly influenced by the viscosity of the bile and the geometric configuration of the cystic duct, i.e., patients with more viscous bile or with a cystic duct containing more baffles or a longer length, have the greatest pressure drop. It is found that the most significant parameter is the diameter of the cystic duct; a 1% decrease in the diameter increases the pressure drop by up to 4.3%. The effects of the baffle height ratio and number of baffles on the pressure drop are reflected in the fact that these effectively change the equivalent diameter and length of the cystic duct. The effect of the Young's modulus on the pressure drop is important only if it is lower than 400 Pa; above this value, a rigid-walled model gives a good estimate of the pressure drop in the system for the parameters studied. PMID- 17408322 TI - Deposition of particles on ocular tissues and formation of Krukenberg spindle, hyphema, and hypopyon. AB - Eye diseases, such as Krukenberg's spindle, hyphema, and hypopyon, are related to the deposition of specific particles such as pigmentary cells, leukocytes, and erythrocytes. These particles are circulated by the aqueous humor (AH) and tend to deposit in regions of low velocities or high resistance. In the present paper, numerical simulations are reported of the AH flow and particle transport, and the particle concentration predictions are qualitatively compared to clinical images. The particle concentration distributions provide an understanding of the likely sources of deposition and the origin of the deposited particles. Pigmentary cells are seen to concentrate in a vertical band on the corneal surface consistent with clinical observations of Krukenberg's spindle. Leukocytes and erythrocytes are seen to collect at the bottom of the anterior chamber similar to the observations made for hypopyon and hyphema. These results confirm the potential of using numerical calculations in order to obtain a better understanding of the particle transport and deposition patterns in the anterior chamber of the eye. PMID- 17408323 TI - Numerical simulation of the urine flow in a stented ureter. AB - When a stent is implanted in a blocked ureter, the urine passing from the kidney to the bladder must traverse a very complicated flow path. That path consists of two parallel passages, one of which is the bore of the stent and the other is the annular space between the external surface of the stent and the inner wall of the ureter. The flow path is further complicated by the presence of numerous pass through holes that are deployed along the length of the stent. These holes allow urine to pass between the annulus and the bore. Further complexity in the pattern of the urine flow occurs because the coiled "pig tails," which hold the stent in place, contain multiple ports for fluid ingress and egress. The fluid flow in a stented ureter has been quantitatively analyzed here for the first time using numerical simulation. The numerical solutions obtained here fully reveal the details of the urine flow throughout the entire stented ureter. It was found that in the absence of blockages, most of the pass-through holes are inactive. Furthermore, only the port in each coiled pig tail that is nearest the stent proper is actively involved in the urine flow. Only in the presence of blockages, which may occur due to encrustation or biofouling, are the numerous pass-through holes activated. The numerical simulations are able to track the urine flow through the pass-through holes as well as adjacent to the blockages. The simulations are also able to provide highly accurate results for the kidney-to bladder urine flow rate. The simulation method presented here constitutes a powerful new tool for rational design of ureteral stents in the future. PMID- 17408324 TI - Application of a microstructural constitutive model of the pulmonary artery to patient-specific studies: validation and effect of orthotropy. AB - We applied a statistical mechanics based microstructural model of pulmonary artery mechanics, developed from our previous studies of rats with pulmonary arterial hypertension (PAH), to patient-specific clinical studies of children with PAH. Our previous animal studies provoked the hypothesis that increased cross-linking density of the molecular chains may be one biological remodeling mechanism by which the PA stiffens in PAH. This study appears to further confirm this hypothesis since varying molecular cross-linking density in the model allows us to simulate the changes in the P-D loops between normotensive and hypertensive conditions reasonably well. The model was combined with patient-specific three dimensional vascular anatomy to obtain detailed information on the topography of stresses and strains within the proximal branches of the pulmonary vasculature. The effect of orthotropy on stressstrain within the main and branch PAs obtained from a patient was explored. This initial study also puts forward important questions that need to be considered before combining the microstructural model with complex patient-specific vascular geometries. PMID- 17408325 TI - Pulsatile blood flow and oxygen transport past a circular cylinder. AB - The fundamental study of blood flow past a circular cylinder filled with an oxygen source is investigated as a building block for an artificial lung. The Casson constitutive equation is used to describe the shear-thinning and yield stress properties of blood. The presence of hemoglobin is also considered. Far from the cylinder, a pulsatile blood flow in the x direction is prescribed, represented by a time periodic (sinusoidal) component superimposed on a steady velocity. The dimensionless parameters of interest for the characterization of the flow and transport are the steady Reynolds number (Re), Womersley parameter (alpha), pulsation amplitude (A), and the Schmidt number (Sc). The Hill equation is used to describe the saturation curve of hemoglobin with oxygen. Two different feed-gas mixtures were considered: pure O(2) and air. The flow and concentration fields were computed for Re=5, 10, and 40, 0< or =A< or =0.75, alpha=0.25, 0.4, and Schmidt number, Sc=1000. The Casson fluid properties result in reduced recirculations (when present) downstream of the cylinder as compared to a Newtonian fluid. These vortices oscillate in size and strength as A and alpha are varied. Hemoglobin enhances mass transport and is especially important for an air feed which is dominated by oxyhemoglobin dispersion near the cylinder. For a pure O(2) feed, oxygen transport in the plasma dominates near the cylinder. Maximum oxygen transport is achieved by operating near steady flow (small A) for both feed-gas mixtures. The time averaged Sherwood number, Sh, is found to be largely influenced by the steady Reynolds number, increasing as Re increases and decreasing with A. Little change is observed with varying alpha for the ranges investigated. The effect of pulsatility on Sh is greater at larger Re. Increasing Re aids transport, but yields a higher cylinder drag force and shear stresses on the cylinder surface which are potentially undesirable. PMID- 17408326 TI - Hip actuations can be used to control bifurcations and chaos in a passive dynamic walking model. AB - We explored how hip joint actuation can be used to control locomotive bifurcations and chaos in a passive dynamic walking model that negotiated a slightly sloped surface (gamma<0.019 rad). With no hip actuation, our passive dynamic walking model was capable of producing a chaotic locomotive pattern when the ramp angle was 0.01839 rad10%. Abnormalities of the long arm of chromosome 3, del(7)(q31q35), trisomy 8, del(11)(q14q23), del(12p) and 20q- could be segregated from their respective IPSS cytogenetic categories and used to develop new cytogenetic subgroups. Clinical parameters, FAB/WHO classification, IPSS score and standard or revised cytogenetic categories were statistically relevant for overall survival (OS) and progression-free intervals (PFI) and were included within five distinct multivariate models compared by the Akaike Information Criterion. To predict OS, the best models included age, WHO classification and standard or revised IPSS cytogenetic categories; to predict PFI, the best model included the same variables and revised cytogenetic categories. In conclusion, (i) the WHO classification was associated with a more homogeneous cytogenetic pattern than the FAB classification, (ii) WHO classification and standard/revised IPSS cytogenetic categories were much more effective than IPSS for predicting MDS clinical outcome, (iii) revised cytogenetic subgroups predicted PFI more effectively than standard categories. PMID- 17408459 TI - Regulation of angiogenesis in the bone marrow of myelodysplastic syndromes transforming to overt leukaemia. AB - To investigate the regulatory mechanisms of angiogenesis in the development of myelodysplastic syndromes (MDS) and its progression to overt leukaemia (OL), bone marrow samples from control, paired samples from MDS patients before and after transformation to OL (MDS --> OL) and de novo acute myeloid leukaemia (AML) were analysed. Immunohistochemical staining showed a significant increase of bone marrow microvascular density (MVD) in MDS and de novo AML compared with controls. Surprisingly, in MDS, MVD significantly decreased upon transformation to OL, which was also significantly lower than the MVD of de novo AML. This evidence was strengthened by the pattern of angiogenic mediator gene expression, confirming the importance of various angiogenic mediators including vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), tumour necrosis factor alpha (TNFalpha), hepatocyte growth factor (HGF) and the angiopoietin family of mediators (Ang-1 and Ang-2) as well as the receptors for angiogenic mediators, such as VEGF receptor 2 (VEGFR2) and the tyrosine kinase receptor, TIE2. By contrast, the anti-angiogenic mediator, transforming growth factor-beta (TGFbeta) exhibited significantly higher expression in the bone marrow of MDS --> OL, indicating the importance of this cytokine as the suppressive factor of angiogenesis in MDS. These findings indicate that the bone marrow microenvironment in MDS --> OL and de novo AML differs remarkably, suggesting the different efficacy of anti-angiogenic therapy between de novo AML and leukaemia secondary to MDS. PMID- 17408460 TI - Stage I/II follicular lymphoma: spread of bcl-2/IgH+ cells in blood and bone marrow from primary site of disease and possibility of clearance after involved field radiotherapy. AB - Stage I/IIA follicular lymphoma (FL) is considered a localised disease that can be adequately treated with radiotherapy alone. Bone marrow (BM) and peripheral blood (PB) involvement in FL was investigated by polymerase chain reaction (PCR) in a series of 24 consecutive patients with histologically revised diagnosis and treated with involved field radiotherapy. Despite the limited stage, Bcl-2/IgH+ cells were found at diagnosis in PB and/or BM of 16 patients (66.6%). After treatment, in 9/15 Bcl-2/IgH positive evaluable patients, a disappearance of Bcl 2/IgH+ cells was observed, which persisted after a median follow-up of 43.5 months (range 11-70) in all but one patient. Quantitative PCR demonstrated the feasibility of clearing PB and BM Bcl-2+ cells after local irradiation of the primary site of the disease only when the basal number of lymphoma cells was <1:100 000. Patients with Bcl-2/IgH+ cells at diagnosis or after treatment had a higher likelihood of relapse. Thus, despite a negative BM biopsy, the majority of localised FL Bcl-2/IgH+ cells were found in the PB and BM. Lymphoma cells can reversibly spread from the affected lymph node to PB and BM and, in a proportion of cases, durably disappear after irradiation. The possibility of a persistent lymphoma cell clearance is proportional to the amount of cells detected at presentation by quantitative PCR. PMID- 17408461 TI - Establishment of a novel childhood acute myeloid leukaemia cell line, KOPM-88, containing partial tandem duplication of the MLL gene and an in vivo model for childhood acute myeloid leukaemia using NOD/SCID mice. AB - MLL gene rearrangement is common in both adult and childhood acute myeloid leukaemia (AML), and its role in oncogenesis has been investigated. While over 50 translocated-partner genes have been identified so far, few studies have detailed the molecular mechanism of partial tandem duplication (PTD) of the MLL gene. The prognostic impact and contribution to leukaemogenesis of MLL-PTD, especially in childhood cases, remain unknown. We have established a novel cell line containing MLL-PTD derived from an 11-year-old patient with AML and designated as KOPM-88. KOPM-88 cells exhibited certain characteristics associated with the myeloid lineage including abundant primary granules in the cytoplasm and the expression of myeloperoxidase. The cell growth of KOPM-88 was cytokine independent but was accelerated by granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. MLL-PTD of exon 2 to exon 6 and exon 2 to exon 8 was revealed using Southern blotting, fluorescence in situ hybridisation, and reverse transcription polymerase chain reaction/DNA sequencing. Furthermore, non-obese diabetic/severe combined immunodeficient mice inoculated with KOPM-88 cells exhibited leukaemic infiltrations in the bone marrow and hemiparalysis because of compression myelopathy. This is the first report of an in vivo animal model exhibiting the systemic involvement of childhood AML containing MLL-PTD. KOPM-88 cells and our murine model may be useful for investigating the pathogenesis of childhood AML associated with MLL gene rearrangement. PMID- 17408462 TI - Cl-IB-MECA inhibits human erythropoiesis. AB - Candidate drugs are being sought for the suppression of human erythropoiesis. Cl IB-MECA [2-chloro-N6-(3-iodobenzyl)-adenosine-5'-N-methyluronamide] is a derivative of adenosine that inhibits the growth of leukaemic cell lines. To determine the effects of Cl-IB-MECA upon erythropoiesis, studies were performed by using an ex vivo culture system of primary human CD34+ cells. Cl-IB-MECA suppressed erythroblast growth and maturation at doses >/=50 mumol/l through a mechanism of cell cycle inhibition and accumulation of cells in the G1/G0 phase. These findings demonstrate that Cl-IB-MECA inhibits human erythropoiesis, and suggest that further consideration of this drug is warranted for patients with erythrocytosis or polycythemia syndromes. PMID- 17408463 TI - The expression of the novel cytotoxic protein granzyme M by large granular lymphocytic leukaemias of both T-cell and NK-cell lineage: an unexpected finding with implications regarding the pathobiology of these disorders. AB - Granzyme M (GrM) is a novel cytotoxic protein normally exclusively expressed by natural killer (NK)-cells and cytotoxic T-cells with innate immune function. As most T-cell granular lymphocytic leukaemias (T-LGL) are thought to be derived from the adaptive immune system it was predicted that T-LGL would be GrM negative. Contrary to this hypothesis, bone marrow biopsy immunohistochemistry revealed that GrM was frequently expressed in both T-LGL (16 / 18) and NK-LGL (6 / 9). These unexpected results suggest commonality between T- and NK-LGL, providing further support to the notion that T-LGL is a disorder of dysregulated, chronically stimulated, adaptive cytotoxic T-cells. PMID- 17408464 TI - Prognostic value of serum free light chain ratio at diagnosis in multiple myeloma. AB - The prognostic value of baseline serum free light chain ratio (sFLCR) was investigated in 94 multiple myeloma (MM) patients. sFLCR was calculated as kappa/lambda or lambda/kappa, depending on the patients' dominating monoclonal light chain. Median baseline sFLCR was 3.57 in kappa-MM patients, 45.09 in lambda MM. 'High' sFLCR (> or = the observed median value for kappa- and lambda-MM respectively) correlated with elevated serum creatinine and lactate dehydrogenase, extensive marrow infiltration and light chain type MM. The 5-year disease-specific survival was 82% and 30% in patients with sFLCR lower than and equal or greater than the median, respectively (P = 0.0001). sFLCR was an independent prognostic factor. PMID- 17408465 TI - Anaemia characterises patients with myelofibrosis harbouring Mpl mutation. AB - The clinical and haematological phenotype of patients with myelofibrosis harbouring MPL(W515L/K) mutation has not been thoroughly investigated. Of 217 myelofibrosis subjects, 18 (8.2%) had an MPL mutation, four of which (22%) co existed with JAK2(V617F) mutation. When compared with MPL wild-type patients, irrespective of JAK2(V617F) status, those with MPL(W515L/K), were more frequently female, were older (61 years vs. 57 years; P = 0.02), presented with more severe anaemia (haemoglobin, 101 g/l vs. 121 g/l; P = 0.002) and were more likely to require regular transfusional support (P = 0.012). These data indicate that MPL mutation in myelofibrosis characterises patients with more severe anaemic phenotype. PMID- 17408466 TI - Second transplant with two unrelated cord blood units for early graft failure after haematopoietic stem cell transplantation. AB - Graft failure (GF) can be a fatal complication following haematopoietic stem cell transplantation (HSCT). We report four patients who developed early GF after unrelated HSCT and who subsequently received a double unrelated cord blood transplant (dUCBT) after reduced-intensity conditioning, at a median 15 d after the decision to perform a second transplant. Neutrophil recovery was observed in all four patients between day +15 and +31 with full donor chimaerism of one unit. Acute GVHD grades II-IV was observed in three patients. Three are alive, between 12 and 25 months after dUCBT. In conclusion, dUCBT is a promising procedure to treat early GF. PMID- 17408467 TI - Iron deprivation blocks multilineage haematopoietic differentiation by inhibiting induction of p21(WAF1/CIP1). AB - Iron is required for the differentiation of HL-60 cells along the monocyte lineage in vitro, reflecting a requirement for iron in the transcriptional induction of the p21(WAF1/CIP1) gene. To determine if the same requirement holds true for differentiation in other cell lineages and for primary human CD34(+) bone marrow precursor cells, we induced granulocyte differentiation by treating HL-60 cells with dimethyl sulphoxide, and erythroid or megakaryocytic differentiation by treating K562 cells with butyrate or phorbol myristate acetate, respectively. Nitro blue tetrazolium reduction, expression of haem, or expression of CD41 was used to assess granulocytic, erythroid, or megakaryocytic differentiation respectively. Purified CD34(+) cells were cultured with granulocyte/macrophage-colony stimulating factor and stem cell factor to induce myelomonocytic differentiation. Iron deprivation was induced by desferrioxamine. p21(WAF1/CIP1) antisense oligonucleotides were used to inhibit p21 expression. Iron deprivation blocked p21 induction as judged by real-time polymerase chain reaction assays. In addition, both iron deprivation and p21 antisense blocked CD34(+) cell differentiation. These observations were not explained by induction of widespread apoptosis under conditions of iron deprivation. We concluded that both iron and functional p21(WAF1/CIP1) are required for in vitro differentiation of human haematopoietic precursors along multiple cell lineages. PMID- 17408468 TI - Genetic polymorphisms associated with priapism in sickle cell disease. AB - Priapism occurs in 30-45% of male patients with sickle cell disease (SCD), but the possible influence of genetic risk factors on the incidence of priapism is not well understood. We examined genetic polymorphisms in 199 unrelated, adult (>18 years), male patients with Hb SS and Hb Sbeta(0)-thalassaemia, 83 (42%) of whom reported a history of priapism. Candidate genes for association with priapism were identified based on their involvement in adhesion, coagulation, inflammation and cell signalling. Additionally, we examined genes involved in nitric oxide biology (NOS2, NOS3, SLC4A1), as well as polymorphisms in the klotho (KL) gene, which has previously been associated with priapism. Strong evidence of association was found for single nucleotide polymorphisms in transforming growth factor-beta receptor, type III (TGFBR3) (rs7526590; P = 0.00058), aquaporin (AQP1) (rs10244884; P = 0.00068), integrin alphav (ITGAV) (rs3768780; P = 0.00090), and the A1 subunit of coagulation factor XIII (F13A1) (hcv1860621; P = 0.00156). Associations with TGFBR3, AQP1, and ITGAV remained significant after adjusting for multiple testing, using the Benjamini-Hochberg procedure. Our data suggest that genes involved in the TGFbeta pathway, coagulation, cell adhesion and cell hydration pathways may be important in risk for priapism. PMID- 17408469 TI - Lenalidomide (Revlimid), in combination with cyclophosphamide and dexamethasone (RCD), is an effective and tolerated regimen for myeloma patients. PMID- 17408470 TI - Acute hepatitis C virus infection assessment among chronic hemodialysis patients in the Southwest Parana State, Brazil. AB - BACKGROUND: Chronic hemodialysis patients are at higher risk for acquiring hepatitis C virus (HCV). The prevalence varies among different countries and hemodialysis centers. Although guidelines for a comprehensive infection control program exist, the nosocomial transmission still accounts for the new cases of infection. The aim of this study was analyze the follow up of newly acquired acute hepatitis C cases, during the period from January 2002 to May 2005, in the Hemodialysis Center, located in the Southwest region of Parana State, Brazil and to analyze the effectiveness of the measures to restrain the appearance of new cases of acute hepatitis C. METHODS: Patients were analyzed monthly with anti-HCV tests and ALT measurements. Patients with ALT elevations were monitored for possible acute hepatitis C. RESULTS: During this period, 32 new cases were identified with acute hepatitis C virus infection. Blood screening showed variable ALT levels preceding the anti-HCV seroconversion. HCV RNA viremia by PCR analysis was intermittently and even negative in some cases. Ten out of 32 patients received 1 mcg/kg dose of pegylated interferon alfa-2b treatment for 24 weeks. All dialysis personnel were re-trained to strictly follow the regulations and recommendations regarding infection control, proper methods to clean and disinfect equipment were reviewed and HCV-positive patients were isolated. CONCLUSION: Laboratory tests results showed variable ALT preceding anti-HCV seroconversion and intermittent viremia. The applied recommendations contributed importantly to restrain the appearance of new cases of acute hepatitis C in this center and the last case was diagnosed in May 2004. PMID- 17408471 TI - The atherogenic and metabolic impact of non-HDL cholesterol versus other lipid sub-components among non-diabetic and diabetic Saudis. AB - BACKGROUND: Several trials from different populations have reported that non-high density lipoprotein cholesterol (non-HDL-C) has more predictive power than low density lipoprotein cholesterol (LDL-C) in detecting coronary heart disease (CHD) and none in any Arab community whose propensity to develop CHD is higher compared to other ethnicities. This study aims to determine and compare the impact of non HDL-C versus other lipid parameters, in predicting coronary heart disease among diabetic versus non-diabetic adult Saudis and identify the lipid parameters which make a significant contribution in the development of coronary heart disease, diabetes mellitus, and metabolic syndrome. 733 adult Saudis were recruited and divided into groups of diabetics and non-diabetics. Each participant completed a questionnaire, underwent physical exam including 12-L ECG, and submitted a fasting blood sample where glucose and lipid parameters were analyzed using routine procedures. RESULTS: 462 subjects (age 45.03 +/- 11.52; BMI 28.91 +/- 6.07) were classified non-diabetics while the remaining 271 (age 52.73 +/- 11.45, BMI 30.15 +/- 6.62) were diabetics. 99 out of 465 (21.3%) of non-diabetics had CHD and 114 out of 271 (52.5%) in the diabetics. Non-HDL cholesterol was the best predictor among the non-diabetics (odds-ratio 2.89, CI 1.10-7.58, p-0.03). Total cholesterol was the highest single predictor for the development of CHD among the lipids (odds-ratio 1.36, CI 0.68-2.71, p-0.39) but HDL-cholesterol although small was significant (odds-ratio 0.52, CI 0.27-0.99, p-0.05). CONCLUSION: This study supports the use of non-HDL cholesterol as the more practical and reliable target for lipid lowering therapy among the Saudi population. PMID- 17408472 TI - Interventions aimed at reducing problems in adult patients discharged from hospital to home: a systematic meta-review. AB - BACKGROUND: Many patients encounter a variety of problems after discharge from hospital and many discharge (planning and support) interventions have been developed and studied. These primary studies have already been synthesized in several literature reviews with conflicting conclusions. We therefore set out a systematic review of the reviews examining discharge interventions. The objective was to synthesize the evidence presented in literature on the effectiveness of interventions aimed to reduce post-discharge problems in adults discharged home from an acute general care hospital. METHODS: A comprehensive search of seventeen literature databases and twenty-five websites was performed for the period 1994 2004 to find relevant reviews. A three-stage inclusion process consisting of initial sifting, checking full-text papers on inclusion criteria, and methodological assessment, was performed independently by two reviewers. Data on effects were synthesized by use of narrative and tabular methods. RESULTS: Fifteen systematic reviews met our inclusion criteria. All reviews had to deal with considerable heterogeneity in interventions, populations and outcomes, making synthesizing and pooling difficult. Although a statistical significant effect was occasionally found, most review authors reached no firm conclusions that the discharge interventions they studied were effective. We found limited evidence that some interventions may improve knowledge of patients, may help in keeping patients at home or may reduce readmissions to hospital. Interventions that combine discharge planning and discharge support tend to lead to the greatest effects. There is little evidence that discharge interventions have an impact on length of stay, discharge destination or dependency at discharge. We found no evidence that discharge interventions have a positive impact on the physical status of patients after discharge, on health care use after discharge, or on costs. CONCLUSION: Based on fifteen high quality systematic reviews, there is some evidence that some interventions may have a positive impact, particularly those with educational components and those that combine pre-discharge and post discharge interventions. However, on the whole there is only limited summarized evidence that discharge planning and discharge support interventions have a positive impact on patient status at hospital discharge, on patient functioning after discharge, on health care use after discharge, or on costs. PMID- 17408473 TI - Jack-of-all-trades, master of none: postgraduate perspectives on interdisciplinary health research in Australia. AB - BACKGROUND: Interdisciplinary health research is increasingly perceived as an expectation of research institutions and funding bodies within Australia. However, little consideration has been given to the extent to which this re orientation has produced a new type of researcher--an interdisciplinary health researcher. DISCUSSION: As cross-enrolled postgraduate research students, we assert that we do not have an intellectual home. Rather, we must forge a virtual intellectual home through the process of bridging disciplines. In this paper we explain that this virtual home affords us the role of 'interlockers' in future health research. The interlocker role privileges a breadth of understandings across disciplines, rather than a depth in one. SUMMARY: We conclude by reiterating that there is an undeniable need for interdisciplinary health research, and that the roles and actions of interdisciplinary health researchers need to be better understood and catered for. We therefore call for increased consideration and discussion concerning the future roles and capacities of interdisciplinary health researchers such as ourselves. PMID- 17408474 TI - On homology searches by protein Blast and the characterization of the age of genes. AB - BACKGROUND: It has been shown in a variety of organisms, including mammals, that genes that appeared recently in evolution, for example orphan genes, evolve faster than older genes. Low functional constraints at the time of origin of novel genes may explain these results. However, this observation has been recently attributed to an artifact caused by the inability of Blast to detect the fastest genes in different eukaryotic genomes. Distinguishing between these two possible explanations would be of great importance for any studies dealing with the taxon distribution of proteins and the origin of novel genes. RESULTS: Here we used simulations of protein sequences to examine the capacity of Blast to detect proteins of diverse evolutionary rates in the different species of an eukaryotic phylogenetic tree that included metazoans, fungi and plants. We simulated the evolution of protein genes with the same evolutionary rates than those observed in functional mammalian genes and with among-site rate heterogeneity. Under these conditions, we found that only a very small percentage of simulated ancestral eukaryotic proteins was affected by the Blast artifact. We show that the good detectability of Blast is due to the heterogeneity of protein evolutionary rates at different sites, since only a small conserved motif in a sequence suffices to detect its homologues. Our results indicate that Blast, at least when applied within eukaryotes, only misses homologues of extremely fast evolving sequences, which are rare in the mammalian genome, as well as sequences evolving homogeneously or pseudogenes. CONCLUSION: Although great care should be exercised in the recognition of remote homologues, most functional mammalian genes can be detected in eukaryotic genomes by Blast. That is, the majority of functional mammalian genes are not as fast as for not being detected in other metazoans, fungi or plants, if they had been present in these organisms. Thus, the correlation previously found between age and rate seems not to be due to a pure Blast artifact, at least for mammals. This may have important implications to understand the mechanisms by which novel genes originate. PMID- 17408476 TI - Infection rates associated with epidural indwelling catheters for seven days or longer: systematic review and meta-analysis. AB - BACKGROUND: To determine infection rate with use of epidural catheters in place for seven days or more. METHODS: Systematic review and pooled analysis of observational studies. RESULTS: Twelve studies with 4,628 patients (median 197 patients) provided information, of which nine (4,334 patients) were published after 1990. Eight studies (3,893 patients) were retrospective, and four studies (735 patients) prospective. Electronic searches identified three studies and searching reference lists nine.There were 257 catheter-related infections in total, of which 211 were superficial and 57 deep, giving rates of 6.1%, 4.6% and 1.2% respectively. Ten of the 12 studies had deep infection rates of 2% or less. The incidence of deep infection was 1 per 2391 days of treatment, or 0.4 per 1000 catheter treatment days. In nine studies (1503 patients), predominantly in cancer, and with average catheter duration of 74 days, the deep infection rate was 2.8%. The proportion of patients with infection of any type was higher in cancer patients with longer catheter duration. Limited numbers of events meant that no reliable estimate of the impact of prospective and retrospective design could be made. There appeared to be a relationship between catheter duration and infection rate from this and other recent estimates. Four of 57 (7%) patients with deep infection died. CONCLUSION: The best estimate is that one person in 35 with an epidural catheter in place for 74 days for relief of cancer pain can be expected to have a deep epidural infection, and that about 1 in 500 may die of infection-related causes. This is a most uncertain estimate given the limited nature of the evidence. PMID- 17408475 TI - Tandem gene arrays in Trypanosoma brucei: comparative phylogenomic analysis of duplicate sequence variation. AB - BACKGROUND: The genome sequence of the protistan parasite Trypanosoma brucei contains many tandem gene arrays. Gene duplicates are created through tandem duplication and are expressed through polycistronic transcription, suggesting that the primary purpose of long, tandem arrays is to increase gene dosage in an environment where individual gene promoters are absent. This report presents the first account of the tandem gene arrays in the T. brucei genome, employing several related genome sequences to establish how variation is created and removed. RESULTS: A systematic survey of tandem gene arrays showed that substantial sequence variation existed across the genome; variation from different regions of an array often produced inconsistent phylogenetic affinities. Phylogenetic relationships of gene duplicates were consistent with concerted evolution being a widespread homogenising force. However, tandem duplicates were not usually identical; therefore, any homogenising effect was coincident with divergence among duplicates. Allelic gene conversion was detected using various criteria and was apparently able to both remove and introduce sequence variation. Tandem arrays containing structural heterogeneity demonstrated how sequence homogenisation and differentiation can occur within a single locus. CONCLUSION: The use of multiple genome sequences in a comparative analysis of tandem gene arrays identified substantial sequence variation among gene duplicates. The distribution of sequence variation is determined by a dynamic balance of conservative and innovative evolutionary forces. Gene trees from various species showed that intraspecific duplicates evolve in concert, perhaps through frequent gene conversion, although this does not prevent sequence divergence, especially where structural heterogeneity physically separates a duplicate from its neighbours. In describing dynamics of sequence variation that have consequences beyond gene dosage, this survey provides a basis for uncovering the hidden functionality within tandem gene arrays in trypanosomatids. PMID- 17408477 TI - Replacement of connexin43 by connexin26 in transgenic mice leads to dysfunctional reproductive organs and slowed ventricular conduction in the heart. AB - BACKGROUND: In order to further distinguish unique from general functions of connexin43, we have generated mice in which the coding region of connexin43 was replaced by that of connexin26. RESULTS: Heterozygous mothers showed impaired mammary gland development responsible for decreased lactation and early postnatal death of the pups which could be partially rescued by wild type foster mothers. Only about 17% of the homozygous connexin43 knock-in connexin26 mice instead of 25% expected according to Mendelian inheritance, were born and only 6% survived to day 21 post partum and longer. Neonatal and adult connexin43 knock-in connexin26 mice exhibited slowed ventricular conduction in their hearts, i.e. similar but delayed electrophysiological abnormalities as connexin43 deficient mice. Furthermore, connexin43 knock-in connexin26 male and female mice were infertile and exhibited hypotrophic gonads. In testes, tubuli seminiferi were developed and spermatogonia as well as some primary spermatocytes were present, but further differentiated stages of spermatogenesis were absent. Ovaries of female connexin43 knock-in connexin26 mice revealed only few follicles and the maturation of follicles was completely impaired. CONCLUSION: The impaired gametogenesis of homozygous males and females can explain their infertility. PMID- 17408478 TI - Problem eating behaviors related to social factors and body weight in preschool children: A longitudinal study. AB - BACKGROUND: Despite the increasing prevalence of overweight/obesity and its association to eating patterns in adolescents and adults, little is known about the relationship between problematic eating behaviours and body weight in the preschool years within the context of various social factors. This research aims to analyze the relationship between social factors, mothers' perceptions of their child's eating behaviour (picky eating and overeating), and body weight in preschool years, in a population-based cohort of preschoolers from Quebec (Canada). METHODS: Analyses were performed on 1498 children from the Longitudinal Study of Child Development in Quebec, a representative sample of children born in 1998 in the Canadian province of Quebec. Eating behaviours (picky eating and overeating) were derived from questionnaires at 2.5, 3.5, and 4.5 years of age. BMI was calculated from children's measured height and weight at 4.5 years. Children's sex and birth weight, mothers' age, immigrant status, smoking status during pregnancy, and education level, family type, annual household income and income sufficiency, the number of overweight/obese parents, children's day-care attendance, and food insufficiency were part of the analysis. Multivariate logistic regressions were used to determine odds ratios for different body weight profiles (underweight, normal weight, at risk of overweight, overweight), and one way analysis-of-variances (ANOVA) allowed for group comparisons of means. RESULTS: The proportion of children reported for each eating behaviour category remained quite stable across the years studied. Picky eating and overeating related to body weight among 4.5-year-old children, even when social and parental factors were accounted for in multivariate analysis. Picky eaters were twice as likely to be underweight at 4.5 years as children who were never picky eaters. Adjusted odds ratios revealed overeaters were 6 times more likely to be overweight at 4.5 years than were children who were never overeaters. CONCLUSION: Given the association between eating behaviours and bodyweight among 4.5-year-old children, particularly among those from less educated, lower income families and younger mothers, health professionals should target parents of children at risk of overweight/obesity and underweight with focussed messages and strategies for the management of emerging problematic eating behaviours. PMID- 17408479 TI - The Edinburgh Postnatal Depression Scale (EPDS): translation and validation study of the Iranian version. AB - BACKGROUND: The Edinburgh Postnatal Depression Scale (EPDS) is a widely used instrument to measure postnatal depression. This study aimed to translate and to test the reliability and validity of the EPDS in Iran. METHODS: The English language version of the EPDS was translated into Persian (Iranian language) and was used in this study. The questionnaire was administered to a consecutive sample of 100 women with normal (n = 50) and caesarean section (n = 50) deliveries at two points in time: 6 to 8 weeks and 12 to 14 weeks after delivery. Statistical analysis was performed to test the reliability and validity of the EPDS. RESULTS: Overall 22% of women at time 1 and 18% at time 2 reported experiencing postpartum depression. In general, the Iranian version of the EPDS was found to be acceptable to almost all women. Cronbach's alpha coefficient (to test reliability) was found to be 0.77 at time 1 and 0.86 at time 2. In addition, test-rest reliability was performed and the intraclass correlation coefficient was found to be 0.80. Validity as performed using known groups comparison showed satisfactory results. The questionnaire discriminated well between sub-groups of women differing in mode of delivery in the expected direction. The factor analysis indicated a three-factor structure that jointly accounted for 58% of the variance. CONCLUSION: This preliminary validation study of the Iranian version of the EPDS proved that it is an acceptable, reliable and valid measure of postnatal depression. It seems that the EPDS not only measures postpartum depression but also may be measuring something more. PMID- 17408480 TI - Genomic and expression analysis of multiple Sry loci from a single Rattus norvegicus Y chromosome. AB - BACKGROUND: Sry is a gene known to be essential for testis determination but is also transcribed in adult male tissues. The laboratory rat, Rattus norvegicus, has multiple Y chromosome copies of Sry while most mammals have only a single copy. DNA sequence comparisons with other rodents with multiple Sry copies are inconsistent in divergence patterns and functionality of the multiple copies. To address hypotheses of divergence, gene conversion and functional constraints, we sequenced Sry loci from a single R. norvegicus Y chromosome from the Spontaneously Hypertensive Rat strain (SHR) and analyzed DNA sequences for homology among copies. Next, to determine whether all copies of Sry are expressed, we developed a modification of the fluorescent marked capillary electrophoresis method to generate three different sized amplification products to identify Sry copies. We applied this fragment analysis method to both genomic DNA and cDNA prepared from mRNA from testis and adrenal gland of adult male rats. RESULTS: Y chromosome fragments were amplified and sequenced using primers that included the entire Sry coding region and flanking sequences. The analysis of these sequences identified six Sry loci on the Y chromosome. These are paralogous copies consistent with a single phylogeny and the divergence between any two copies is less than 2%. All copies have a conserved reading frame and amino acid sequence consistent with function. Fragment analysis of genomic DNA showed close approximations of experimental with predicted values, validating the use of this method to identify proportions of each copy. Using the fragment analysis procedure with cDNA samples showed the Sry copies expressed were significantly different from the genomic distribution (testis p < 0.001, adrenal gland p < 0.001), and the testis and adrenal copy distribution in the transcripts were also significantly different from each other (p < 0.001). Total Sry transcript expression, analyzed by real-time PCR, showed significantly higher levels of Sry in testis than adrenal gland (p, 0.001). CONCLUSION: The SHR Y chromosome contains at least 6 full length copies of the Sry gene. These copies have a conserved coding region and conserved amino acid sequence. The pattern of divergence is not consistent with gene conversion as the mechanism for this conservation. Expression studies show multiple copies expressed in the adult male testis and adrenal glands, with tissue specific differences in expression patterns. Both the DNA sequence analysis and RNA transcript expression analysis are consistent with more than one copy having function and selection preventing divergence although we have no functional evidence. PMID- 17408481 TI - Gene expression profiles in rat brain disclose CNS signature genes and regional patterns of functional specialisation. AB - BACKGROUND: The mammalian brain is divided into distinct regions with structural and neurophysiological differences. As a result, gene expression is likely to vary between regions in relation to their cellular composition and neuronal function. In order to improve our knowledge and understanding of regional patterns of gene expression in the CNS, we have generated a global map of gene expression in selected regions of the adult rat brain (frontomedial-, temporal- and occipital cortex, hippocampus, striatum and cerebellum; both right and left sides) as well as in three major non-neural tissues (spleen, liver and kidney) using the Applied Biosystems Rat Genome Survey Microarray. RESULTS: By unsupervised hierarchical clustering, we found that the transcriptome within a region was highly conserved among individual rats and that there were no systematic differences between the two hemispheres (right versus left side). Further, we identified distinct sets of genes showing significant regional enrichment. Functional annotation of each of these gene sets clearly reflected several important physiological features of the region in question, including synaptic transmission within the cortex, neurogenesis in hippocampus and G protein-mediated signalling in striatum. In addition, we were able to reveal potentially new regional features, such as mRNA transcription- and neurogenesis annotated activities in cerebellum and differential use of glutamate signalling between regions. Finally, we determined a set of 'CNS-signature' genes that uncover characteristics of several common neuronal processes in the CNS, with marked over-representation of specific features of synaptic transmission, ion transport and cell communication, as well as numerous novel unclassified genes. CONCLUSION: We have generated a global map of gene expression in the rat brain and used this to determine functional processes and pathways that have a regional preference or ubiquitous distribution within the CNS, respectively. The existence of shared specialised neuronal activities in CNS is interesting in a context of potential functional redundancy, and future studies should further explore the overall characteristics of CNS-specific versus region-specific gene profiles in the brain. PMID- 17408482 TI - Amelogenesis imperfecta. AB - Amelogenesis imperfecta (AI) represents a group of developmental conditions, genomic in origin, which affect the structure and clinical appearance of enamel of all or nearly all the teeth in a more or less equal manner, and which may be associated with morphologic or biochemical changes elsewhere in the body. The prevalence varies from 1:700 to 1:14,000, according to the populations studied. The enamel may be hypoplastic, hypomineralised or both and teeth affected may be discoloured, sensitive or prone to disintegration. AI exists in isolation or associated with other abnormalities in syndromes. It may show autosomal dominant, autosomal recessive, sex-linked and sporadic inheritance patterns. In families with an X-linked form it has been shown that the disorder may result from mutations in the amelogenin gene, AMELX. The enamelin gene, ENAM, is implicated in the pathogenesis of the dominant forms of AI. Autosomal recessive AI has been reported in families with known consanguinity. Diagnosis is based on the family history, pedigree plotting and meticulous clinical observation. Genetic diagnosis is presently only a research tool. The condition presents problems of socialisation, function and discomfort but may be managed by early vigorous intervention, both preventively and restoratively, with treatment continued throughout childhood and into adult life. In infancy, the primary dentition may be protected by the use of preformed metal crowns on posterior teeth. The longer term care involves either crowns or, more frequently these days, adhesive, plastic restorations. PMID- 17408483 TI - Melatonin the "light of night" in human biology and adolescent idiopathic scoliosis. AB - Melatonin "the light of night" is secreted from the pineal gland principally at night. The hormone is involved in sleep regulation, as well as in a number of other cyclical bodily activities and circadian rhythm in humans. Melatonin is exclusively involved in signalling the 'time of day' and 'time of year' (hence considered to help both clock and calendar functions) to all tissues and is thus considered to be the body's chronological pacemaker or 'Zeitgeber'. The last decades melatonin has been used as a therapeutic chemical in a large spectrum of diseases, mainly in sleep disturbances and tumours and may play a role in the biologic regulation of mood, affective disorders, cardiovascular system, reproduction and aging. There are few papers regarding melatonin and its role in adolescent idiopathic scoliosis (AIS). Melatonin may play a role in the pathogenesis of scoliosis (neuroendocrine hypothesis) but at present, the data available cannot clearly support this hypothesis. Uncertainties and doubts still surround the role of melatonin in human physiology and pathophysiology and future research is needed. PMID- 17408484 TI - Quantifying geocode location error using GIS methods. AB - BACKGROUND: The Metropolitan Atlanta Congenital Defects Program (MACDP) collects maternal address information at the time of delivery for infants and fetuses with birth defects. These addresses have been geocoded by two independent agencies: (1) the Georgia Division of Public Health Office of Health Information and Policy (OHIP) and (2) a commercial vendor. Geographic information system (GIS) methods were used to quantify uncertainty in the two sets of geocodes using orthoimagery and tax parcel datasets. METHODS: We sampled 599 infants and fetuses with birth defects delivered during 1994-2002 with maternal residence in either Fulton or Gwinnett County. Tax parcel datasets were obtained from the tax assessor's offices of Fulton and Gwinnett County. High-resolution orthoimagery for these counties was acquired from the U.S. Geological Survey. For each of the 599 addresses we attempted to locate the tax parcel corresponding to the maternal address. If the tax parcel was identified the distance and the angle between the geocode and the residence were calculated. We used simulated data to characterize the impact of geocode location error. In each county 5,000 geocodes were generated and assigned their corresponding Census 2000 tract. Each geocode was then displaced at a random angle by a random distance drawn from the distribution of observed geocode location errors. The census tract of the displaced geocode was determined. We repeated this process 5,000 times and report the percentage of geocodes that resolved into incorrect census tracts. RESULTS: Median location error was less than 100 meters for both OHIP and commercial vendor geocodes; the distribution of angles appeared uniform. Median location error was approximately 35% larger in Gwinnett (a suburban county) relative to Fulton (a county with urban and suburban areas). Location error occasionally caused the simulated geocodes to be displaced into incorrect census tracts; the median percentage of geocodes resolving into incorrect census tracts ranged between 4.5% and 5.3%, depending upon the county and geocoding agency. CONCLUSION: Geocode location uncertainty can be estimated using tax parcel databases in a GIS. This approach is a viable alternative to global positioning system field validation of geocodes. PMID- 17408485 TI - Nitrogen fixation in eukaryotes--new models for symbiosis. AB - BACKGROUND: Nitrogen, a component of many bio-molecules, is essential for growth and development of all organisms. Most nitrogen exists in the atmosphere, and utilisation of this source is important as a means of avoiding nitrogen starvation. However, the ability to fix atmospheric nitrogen via the nitrogenase enzyme complex is restricted to some bacteria. Eukaryotic organisms are only able to obtain fixed nitrogen through their symbiotic interactions with nitrogen fixing prokaryotes. These symbioses involve a variety of host organisms, including animals, plants, fungi and protists. RESULTS: We have compared the morphological, physiological and molecular characteristics of nitrogen fixing symbiotic associations of bacteria and their diverse hosts. Special features of the interaction, e.g. vertical transmission of symbionts, grade of dependency of partners and physiological modifications have been considered in terms of extent of co-evolution and adaptation. Our findings are that, despite many adaptations enabling a beneficial partnership, most symbioses for molecular nitrogen fixation involve facultative interactions. However, some interactions, among them endosymbioses between cyanobacteria and diatoms, show characteristics that reveal a more obligate status of co-evolution. CONCLUSION: Our review emphasises that molecular nitrogen fixation, a driving force for interactions and co-evolution of different species, is a widespread phenomenon involving many different organisms and ecosystems. The diverse grades of symbioses, ranging from loose associations to highly specific intracellular interactions, might themselves reflect the range of potential evolutionary fates for symbiotic partnerships. These include the extreme evolutionary modifications and adaptations that have accompanied the formation of organelles in eukaryotic cells: plastids and mitochondria. However, age and extensive adaptation of plastids and mitochondria complicate the investigation of processes involved in the transition of symbionts to organelles. Extant lineages of symbiotic associations for nitrogen fixation show diverse grades of adaptation and co-evolution, thereby representing different stages of symbiont-host interaction. In particular cyanobacterial associations with protists, like the Rhopalodia gibba-spheroid body symbiosis, could serve as important model systems for the investigation of the complex mechanisms underlying organelle evolution. PMID- 17408486 TI - Integration of Arabidopsis thaliana stress-related transcript profiles, promoter structures, and cell-specific expression. AB - BACKGROUND: Arabidopsis thaliana transcript profiles indicate effects of abiotic and biotic stresses and tissue-specific and cell-specific gene expression. Organizing these datasets could reveal the structure and mechanisms of responses and crosstalk between pathways, and in which cells the plants perceive, signal, respond to, and integrate environmental inputs. RESULTS: We clustered Arabidopsis transcript profiles for various treatments, including abiotic, biotic, and chemical stresses. Ubiquitous stress responses in Arabidopsis, similar to those of fungi and animals, employ genes in pathways related to mitogen-activated protein kinases, Snf1-related kinases, vesicle transport, mitochondrial functions, and the transcription machinery. Induced responses to stresses are attributed to genes whose promoters are characterized by a small number of regulatory motifs, although secondary motifs were also apparent. Most genes that are downregulated by stresses exhibited distinct tissue-specific expression patterns and appear to be under developmental regulation. The abscisic acid dependent transcriptome is delineated in the cluster structure, whereas functions that are dependent on reactive oxygen species are widely distributed, indicating that evolutionary pressures confer distinct responses to different stresses in time and space. Cell lineages in roots express stress-responsive genes at different levels. Intersections of stress-responsive and cell-specific profiles identified cell lineages affected by abiotic stress. CONCLUSION: By analyzing the stress-dependent expression profile, we define a common stress transcriptome that apparently represents universal cell-level stress responses. Combining stress dependent and tissue-specific and cell-specific expression profiles, and Arabidopsis 5'-regulatory DNA sequences, we confirm known stress-related 5' cis elements on a genome-wide scale, identify secondary motifs, and place the stress response within the context of tissues and cell lineages in the Arabidopsis root. PMID- 17408487 TI - Genetic microheterogeneity and phenotypic variation of Helicobacter pylori arginase in clinical isolates. AB - BACKGROUND: Clinical isolates of the gastric pathogen Helicobacter pylori display a high level of genetic macro- and microheterogeneity, featuring a panmictic, rather than clonal structure. The ability of H. pylori to survive the stomach acid is due, in part, to the arginase-urease enzyme system. Arginase (RocF) hydrolyzes L-arginine to L-ornithine and urea, and urease hydrolyzes urea to carbon dioxide and ammonium, which can neutralize acid. RESULTS: The degree of variation in arginase was explored at the DNA sequence, enzyme activity and protein expression levels. To this end, arginase activity was measured from 73 minimally-passaged clinical isolates and six laboratory-adapted strains of H. pylori. The rocF gene from 21 of the strains was cloned into genetically stable E. coli and the enzyme activities measured. Arginase activity was found to substantially vary (>100-fold) in both different H. pylori strains and in the E. coli model. Western blot analysis revealed a positive correlation between activity and amount of protein expressed in most H. pylori strains. Several H. pylori strains featured altered arginase activity upon in vitro passage. Pairwise alignments of the 21 rocF genes plus strain J99 revealed extensive microheterogeneity in the promoter region and 3' end of the rocF coding region. Amino acid S232, which was I232 in the arginase-negative clinical strain A2, was critical for arginase activity. CONCLUSION: These studies demonstrated that H. pylori arginase exhibits extensive genotypic and phenotypic variation which may be used to understand mechanisms of microheterogeneity in H. pylori. PMID- 17408488 TI - Types of psychosocial job demands and adverse events due to dental mismanagement: a cross sectional study. AB - BACKGROUND: A harsh work environment including psychosocial job demands might cause adverse events due to medical mismanagement, but the association has not been explored. The purpose of the study was to investigate whether some types of psychosocial job demands are associated with adverse events due to dental mismanagement experienced by general dental practitioners. METHODS: A self administered questionnaire was mailed to members of a local branch of the Japan dental association. A total of 261 dental practitioners responded anonymously (response rate 53%). Psychosocial job demands were measured by a Japanese version of the Copenhagen Psychosocial Questionnaire, which comprises five sub-scales: quantitative demands, cognitive demands, emotional demands, demands for hiding emotions, and sensorial demands. The outcome was defined according to whether the respondent's patients experienced one of the following adverse events due to dental mismanagement at least once during the previous one year: dropping of dental instrument or broken injection needle, soft tissue or nerve injury, accidental bleeding, loss of a tooth root into the maxillary sinus, and emphysema. Associations between each demand index and experience of adverse events were examined by logistic regression analyses adjusting for potential confounders. RESULTS: Emotional demands and sensorial demands were significantly associated with the experience of adverse events (odds ratio = 3.9 for each). Other than the indices, male gender, younger age, practice alone, many dental chairs (five or more), and many patients (30 or more per day) were the risks. Working hours per week and number of paramedical staff had no significant associations. CONCLUSION: Emotional and sensorial job demands are a potential target for the reduction of adverse events due to dental mismanagement. PMID- 17408489 TI - The gender specific frequency of risk factor and CHD diagnoses prior to incident MI: a community study. AB - BACKGROUND: CHD is a chronic disease often present years prior to incident AMI. Earlier recognition of CHD may be associated with higher levels of recognition and treatment of CHD risk factors that may delay incident AMI. To assess timing of CHD and CHD risk factor diagnoses prior to incident AMI. METHODS: This is a 10 year population based medical record review study that included all medical care providers in Olmsted County, Minnesota for all women and a sample of men residing in Olmsted County, MN with confirmed incident AMI between 1995 and 2000. RESULTS: All medical care for the 10 years prior to incident AMI was reviewed for 150 women and 148 men (38% sample) in Olmsted County, MN. On average, women were older than men at the time of incident AMI (74.7 versus 65.9 years, p < 0.0001). 30.4% of the men and 52.0% of the women received diagnoses of CHD prior to incident AMI (p = 0.0002). Unrecognized and untreated CHD risk factors were present in both men (45% of men 5 years prior to AMI) and women (22% of women 5 years prior to first AMI), more common in men and those without a diagnosis of CHD prior to incident AMI (p < 0.0001). CONCLUSION: A CHD diagnosis prior to incident AMI is associated with higher rates of recognition and treatment of CHD risk factors suggesting that diagnosing CHD prior to AMI enhances opportunities to lower the risk of future CHD events. PMID- 17408490 TI - Colony size measurement of the yeast gene deletion strains for functional genomics. AB - BACKGROUND: Numerous functional genomics approaches have been developed to study the model organism yeast, Saccharomyces cerevisiae, with the aim of systematically understanding the biology of the cell. Some of these techniques are based on yeast growth differences under different conditions, such as those generated by gene mutations, chemicals or both. Manual inspection of the yeast colonies that are grown under different conditions is often used as a method to detect such growth differences. RESULTS: Here, we developed a computerized image analysis system called Growth Detector (GD), to automatically acquire quantitative and comparative information for yeast colony growth. GD offers great convenience and accuracy over the currently used manual growth measurement method. It distinguishes true yeast colonies in a digital image and provides an accurate coordinate oriented map of the colony areas. Some post-processing calculations are also conducted. Using GD, we successfully detected a genetic linkage between the molecular activity of the plant-derived antifungal compound berberine and gene expression components, among other cellular processes. A novel association for the yeast mek1 gene with DNA damage repair was also identified by GD and confirmed by a plasmid repair assay. The results demonstrate the usefulness of GD for yeast functional genomics research. CONCLUSION: GD offers significant improvement over the manual inspection method to detect relative yeast colony size differences. The speed and accuracy associated with GD makes it an ideal choice for large-scale functional genomics investigations. PMID- 17408491 TI - A molecular phylogeny of Dorylus army ants provides evidence for multiple evolutionary transitions in foraging niche. AB - BACKGROUND: Army ants are the prime arthropod predators in tropical forests, with huge colonies and an evolutionary derived nomadic life style. Five of the six recognized subgenera of Old World Dorylus army ants forage in the soil, whereas some species of the sixth subgenus (Anomma) forage in the leaf-litter and some as conspicuous swarm raiders on the forest floor and in the lower vegetation (the infamous driver ants). Here we use a combination of nuclear and mitochondrial DNA sequences to reconstruct the phylogeny of the Dorylus s.l. army ants and to infer the evolutionary transitions in foraging niche and associated morphological adaptations. RESULTS: Underground foraging is basal and gave rise to leaf-litter foraging. Leaf-litter foraging in turn gave rise to two derived conditions: true surface foraging (the driver ants) and a reversal to subterranean foraging (a clade with most of the extant Dorylus s.s. species). This means that neither the subgenus Anomma nor Dorylus s.s. is monophyletic, and that one of the Dorylus s.s. lineages adopted subterranean foraging secondarily. We show that this latter group evolved a series of morphological adaptations to underground foraging that are remarkably convergent to the basal state. CONCLUSION: The evolutionary transitions in foraging niche were more complex than previously thought, but our comparative analysis of worker morphology lends strong support to the contention that particular foraging niches have selected for very specific worker morphologies. The surprising reversal to underground foraging is therefore a striking example of convergent morphological evolution. PMID- 17408492 TI - Contribution for new genetic markers of rheumatoid arthritis activity and severity: sequencing of the tumor necrosis factor-alpha gene promoter. AB - The objective of this study was to assess whether clinical measures of rheumatoid arthritis activity and severity were influenced by tumor necrosis factor-alpha (TNF-alpha) promoter genotype/haplotype markers. Each patient's disease activity was assessed by the disease activity score using 28 joint counts (DAS28) and functional capacity by the Health Assessment Questionnaire (HAQ) score. Systemic manifestations, radiological damage evaluated by the Sharp/van der Heijde (SvdH) score, disease-modifying anti-rheumatic drug use, joint surgeries, and work disability were also assessed. The promoter region of the TNF-alpha gene, between nucleotides -1,318 and +49, was sequenced using an automated platform. Five hundred fifty-four patients were evaluated and genotyped for 10 single-nucleotide polymorphism (SNP) markers, but 5 of these markers were excluded due to failure to fall within Hardy-Weinberg equilibrium or to monomorphism. Patients with more than 10 years of disease duration (DD) presented significant associations between the -857 SNP and systemic manifestations, as well as joint surgeries. Associations were also found between the -308 SNP and work disability in patients with more than 2 years of DD and radiological damage in patients with less than 10 years of DD. A borderline effect was found between the -238 SNP and HAQ score and radiological damage in patients with 2 to 10 years of DD. An association was also found between haplotypes and the SvdH score for those with more than 10 years of DD. An association was found between some TNF-alpha promoter SNPs and systemic manifestations, radiological progression, HAQ score, work disability, and joint surgeries, particularly in some classes of DD and between haplotypes and radiological progression for those with more than 10 years of DD. PMID- 17408493 TI - Risk factor studies of age-at-onset in a sample ascertained for Parkinson disease affected sibling pairs: a cautionary tale. AB - An association between exposure to a risk factor and age-at-onset of disease may reflect an effect on the rate of disease occurrence or an acceleration of the disease process. The difference in age-at-onset arising from case-only studies, however, may also reflect secular trends in the prevalence of exposure to the risk factor. Comparisons of age-at-onset associated with risk factors are commonly performed in case series enrolled for genetic linkage analysis of late onset diseases. We describe how the results of age-at-onset studies of environmental risk factors reflect the underlying structure of the source population, rather than an association with age-at-onset, by contrasting the effects of coffee drinking and cigarette smoking on Parkinson disease age-at onset with the effects on age-at-enrollment in a population based study sample. Despite earlier evidence to suggest a protective association of coffee drinking and cigarette smoking with Parkinson disease risk, the age-at-onset results are comparable to the patterns observed in the population sample, and thus a causal inference from the age-at-onset effect may not be justified. Protective effects of multivitamin use on PD age-at-onset are also shown to be subject to a bias from the relationship between age and multivitamin initiation. Case-only studies of age-at-onset must be performed with an appreciation for the association between risk factors and age and ageing in the source population. PMID- 17408494 TI - Cost-effective design of economic instruments in nutrition policy. AB - This paper addresses the potential for using economic regulation, e.g. taxes or subsidies, as instruments to combat the increasing problems of inappropriate diets, leading to health problems such as obesity, diabetes 2, cardiovascular diseases etc. in most countries. Such policy measures may be considered as alternatives or supplements to other regulation instruments, including information campaigns, bans or enhancement of technological solutions to the problems of obesity or related diseases. 7 different food tax and subsidy instruments or combinations of instruments are analysed quantitatively. The analyses demonstrate that the average cost-effectiveness with regard to changing the intake of selected nutritional variables can be improved by 10-30 per cent if taxes/subsidies are targeted against these nutrients, compared with targeting selected food categories. Finally, the paper raises a range of issues, which need to be investigated further, before firm conclusions about the suitability of economic instruments in nutrition policy can be drawn. PMID- 17408495 TI - Selective reconstitution of liver cholesterol biosynthesis promotes lung maturation but does not prevent neonatal lethality in Dhcr7 null mice. AB - BACKGROUND: Targeted disruption of the murine 3beta-hydroxysterol-Delta7 reductase gene (Dhcr7), an animal model of Smith-Lemli-Opitz syndrome, leads to loss of cholesterol synthesis and neonatal death that can be partially rescued by transgenic replacement of DHCR7 expression in brain during embryogenesis. To gain further insight into the role of non-brain tissue cholesterol deficiency in the pathophysiology, we tested whether the lethal phenotype could be abrogated by selective transgenic complementation with DHCR7 expression in the liver. RESULTS: We generated mice that carried a liver-specific human DHCR7 transgene whose expression was driven by the human apolipoprotein E (ApoE) promoter and its associated liver-specific enhancer. These mice were then crossed with Dhcr7+/- mutants to generate Dhcr7-/- mice bearing a human DHCR7 transgene. Robust hepatic transgene expression resulted in significant improvement of cholesterol homeostasis with cholesterol concentrations increasing to 80~90 % of normal levels in liver and lung. Significantly, cholesterol deficiency in brain was not altered. Although late gestational lung sacculation defect reported previously was significantly improved, there was no parallel increase in postnatal survival in the transgenic mutant mice. CONCLUSION: The reconstitution of DHCR7 function selectively in liver induced a significant improvement of cholesterol homeostasis in non-brain tissues, but failed to rescue the neonatal lethality of Dhcr7 null mice. These results provided further evidence that CNS defects caused by Dhcr7 null likely play a major role in the lethal pathogenesis of Dhcr7-/- mice, with the peripheral organs contributing the morbidity. PMID- 17408496 TI - Evaluation of toxicity of the mycotoxin citrinin using yeast ORF DNA microarray and Oligo DNA microarray. AB - BACKGROUND: Mycotoxins are fungal secondary metabolites commonly present in feed and food, and are widely regarded as hazardous contaminants. Citrinin, one of the very well known mycotoxins that was first isolated from Penicillium citrinum, is produced by more than 10 kinds of fungi, and is possibly spread all over the world. However, the information on the action mechanism of the toxin is limited. Thus, we investigated the citrinin-induced genomic response for evaluating its toxicity. RESULTS: Citrinin inhibited growth of yeast cells at a concentration higher than 100 ppm. We monitored the citrinin-induced mRNA expression profiles in yeast using the ORF DNA microarray and Oligo DNA microarray, and the expression profiles were compared with those of the other stress-inducing agents. Results obtained from both microarray experiments clustered together, but were different from those of the mycotoxin patulin. The oxidative stress response genes--AADs, FLR1, OYE3, GRE2, and MET17--were significantly induced. In the functional category, expression of genes involved in "metabolism", "cell rescue, defense and virulence", and "energy" were significantly activated. In the category of "metabolism", genes involved in the glutathione synthesis pathway were activated, and in the category of "cell rescue, defense and virulence", the ABC transporter genes were induced. To alleviate the induced stress, these cells might pump out the citrinin after modification with glutathione. While, the citrinin treatment did not induce the genes involved in the DNA repair. CONCLUSION: Results from both microarray studies suggest that citrinin treatment induced oxidative stress in yeast cells. The genotoxicity was less severe than the patulin, suggesting that citrinin is less toxic than patulin. The reproducibility of the expression profiles was much better with the Oligo DNA microarray. However, the Oligo DNA microarray did not completely overcome cross hybridization. PMID- 17408497 TI - Bursaries, writing grants and fellowships: a strategy to develop research capacity in primary health care. AB - BACKGROUND: General practitioners and other primary health care professionals are often the first point of contact for patients requiring health care. Identifying, understanding and linking current evidence to best practice can be challenging and requires at least a basic understanding of research principles and methodologies. However, not all primary health care professionals are trained in research or have research experience. With the aim of enhancing research skills and developing a research culture in primary health care, University Departments of General Practice and Rural Health have been supported since 2000 by the Australian Government funded 'Primary Health Care Research Evaluation and Development (PHCRED) Strategy'. A small grant funding scheme to support primary health care practitioners was implemented through the PHCRED program at Flinders University in South Australia between 2002 and 2005. The scheme incorporated academic mentors and three types of funding support: bursaries, writing grants and research fellowships. This article describes outcomes of the funding scheme and contributes to the debate surrounding the effectiveness of funding schemes as a means of building research capacity. METHODS: Funding recipients who had completed their research were invited to participate in a semi-structured 40 minute telephone interview. Feedback was sought on acquisition of research skills, publication outcomes, development of research capacity, confidence and interest in research, and perception of research. Data were also collected on demographics, research topics, and time needed to complete planned activities. RESULTS: The funding scheme supported 24 bursaries, 11 writing grants, and three research fellows. Nearly half (47%) of all grant recipients were allied health professionals, followed by general practitioners (21%). The majority (70%) were novice and early career researchers. Eighty-nine percent of the grant recipients were interviewed. Capacity, confidence, and level of research skills in ten core areas were generally considered to have improved as a result of the award. More than half (53%) had presented their research and 32% had published or submitted an article in a peer-reviewed journal. CONCLUSION: A small grant and mentoring scheme through a University Department can effectively enhance research skills, confidence, output, and interest in research of primary health care practitioners. PMID- 17408498 TI - Synthesis of the oxysterol, 24(S), 25-epoxycholesterol, parallels cholesterol production and may protect against cellular accumulation of newly-synthesized cholesterol. AB - AIM: The effects of 24(S),25-epoxycholesterol (24,25EC) on aspects of cholesterol homeostasis is well-documented. When added to cells, 24,25EC decreases cholesterol synthesis and up-regulates cholesterol efflux genes, including ABCA1. Synthesis of 24,25EC occurs in a shunt of the mevalonate pathway which also produces cholesterol. Therefore, 24,25EC synthesis should be subject to the same negative feedback regulation as cholesterol synthesis. To date, no role has been ascribed to 24,25EC in light of the fact that increased accumulation of cholesterol should decrease formation of this oxysterol through feedback inhibition. This leads to the intriguing paradox: why inhibit production of an apparently important regulator of cholesterol homeostasis when it is needed most? METHODS: We used a combination of pharmacological and genetic approaches in Chinese Hamster Ovary cell-lines to investigate this paradox. Endogenous synthesis of 24,25EC was manipulated using partial inhibition of the enzyme, Oxidosqualene Cyclase. Changes in cholesterol and 24,25EC synthesis were determined using metabolic labelling with [1-14C]-acetate, thin-layer chromatography and phosphorimaging. Transcriptional effects mediated via SREBP and LXR were analysed by luciferase reporter assays. RESULTS: We showed that cholesterol addition to cells lead to a rapid and preferential inhibition of 24,25EC synthesis. Addition of 24,25EC resulted in parallel inhibition of 24,25EC and cholesterol synthesis. Furthermore, we used a variety of approaches to examine the relationship between cholesterol and 24,25EC synthesis, including cell-lines with different rates of cholesterol synthesis, varying cholesterol synthetic rates by pre-treatment with a statin, or lipoprotein cholesterol loading of macrophages. In all cases, we showed that 24,25EC synthesis faithfully tracked cholesterol synthesis. Moreover, changes in 24,25EC synthesis exerted downstream effects, reducing SREBP transcriptional activity whilst increasing ABCA1 and LXR transcriptional activity. CONCLUSION: Our results show that 24,25EC synthesis parallels cholesterol synthesis, consistent with this oxysterol functioning as a safety valve to protect against the accumulation of newly synthesised cholesterol (as opposed to exogenously-derived cholesterol). Considering that 24,25EC is capable of being produced in all cholesterogenic cells, we propose that production of 24,25EC may represent a ubiquitous defence mechanism. PMID- 17408499 TI - Simultaneous clustering of gene expression data with clinical chemistry and pathological evaluations reveals phenotypic prototypes. AB - BACKGROUND: Commonly employed clustering methods for analysis of gene expression data do not directly incorporate phenotypic data about the samples. Furthermore, clustering of samples with known phenotypes is typically performed in an informal fashion. The inability of clustering algorithms to incorporate biological data in the grouping process can limit proper interpretation of the data and its underlying biology. RESULTS: We present a more formal approach, the modk prototypes algorithm, for clustering biological samples based on simultaneously considering microarray gene expression data and classes of known phenotypic variables such as clinical chemistry evaluations and histopathologic observations. The strategy involves constructing an objective function with the sum of the squared Euclidean distances for numeric microarray and clinical chemistry data and simple matching for histopathology categorical values in order to measure dissimilarity of the samples. Separate weighting terms are used for microarray, clinical chemistry and histopathology measurements to control the influence of each data domain on the clustering of the samples. The dynamic validity index for numeric data was modified with a category utility measure for determining the number of clusters in the data sets. A cluster's prototype, formed from the mean of the values for numeric features and the mode of the categorical values of all the samples in the group, is representative of the phenotype of the cluster members. The approach is shown to work well with a simulated mixed data set and two real data examples containing numeric and categorical data types. One from a heart disease study and another from acetaminophen (an analgesic) exposure in rat liver that causes centrilobular necrosis. CONCLUSION: The modk-prototypes algorithm partitioned the simulated data into clusters with samples in their respective class group and the heart disease samples into two groups (sick and buff denoting samples having pain type representative of angina and non-angina respectively) with an accuracy of 79%. This is on par with, or better than, the assignment accuracy of the heart disease samples by several well-known and successful clustering algorithms. Following modk-prototypes clustering of the acetaminophen-exposed samples, informative genes from the cluster prototypes were identified that are descriptive of, and phenotypically anchored to, levels of necrosis of the centrilobular region of the rat liver. The biological processes cell growth and/or maintenance, amine metabolism, and stress response were shown to discern between no and moderate levels of acetaminophen-induced centrilobular necrosis. The use of well-known and traditional measurements directly in the clustering provides some guarantee that the resulting clusters will be meaningfully interpretable. PMID- 17408500 TI - In silico pathway reconstruction: Iron-sulfur cluster biogenesis in Saccharomyces cerevisiae. AB - BACKGROUND: Current advances in genomics, proteomics and other areas of molecular biology make the identification and reconstruction of novel pathways an emerging area of great interest. One such class of pathways is involved in the biogenesis of Iron-Sulfur Clusters (ISC). RESULTS: Our goal is the development of a new approach based on the use and combination of mathematical, theoretical and computational methods to identify the topology of a target network. In this approach, mathematical models play a central role for the evaluation of the alternative network structures that arise from literature data-mining, phylogenetic profiling, structural methods, and human curation. As a test case, we reconstruct the topology of the reaction and regulatory network for the mitochondrial ISC biogenesis pathway in S. cerevisiae. Predictions regarding how proteins act in ISC biogenesis are validated by comparison with published experimental results. For example, the predicted role of Arh1 and Yah1 and some of the interactions we predict for Grx5 both matches experimental evidence. A putative role for frataxin in directly regulating mitochondrial iron import is discarded from our analysis, which agrees with also published experimental results. Additionally, we propose a number of experiments for testing other predictions and further improve the identification of the network structure. CONCLUSION: We propose and apply an iterative in silico procedure for predictive reconstruction of the network topology of metabolic pathways. The procedure combines structural bioinformatics tools and mathematical modeling techniques that allow the reconstruction of biochemical networks. Using the Iron Sulfur cluster biogenesis in S. cerevisiae as a test case we indicate how this procedure can be used to analyze and validate the network model against experimental results. Critical evaluation of the obtained results through this procedure allows devising new wet lab experiments to confirm its predictions or provide alternative explanations for further improving the models. PMID- 17408501 TI - Reconstructing gene-regulatory networks from time series, knock-out data, and prior knowledge. AB - BACKGROUND: Cellular processes are controlled by gene-regulatory networks. Several computational methods are currently used to learn the structure of gene regulatory networks from data. This study focusses on time series gene expression and gene knock-out data in order to identify the underlying network structure. We compare the performance of different network reconstruction methods using synthetic data generated from an ensemble of reference networks. Data requirements as well as optimal experiments for the reconstruction of gene regulatory networks are investigated. Additionally, the impact of prior knowledge on network reconstruction as well as the effect of unobserved cellular processes is studied. RESULTS: We identify linear Gaussian dynamic Bayesian networks and variable selection based on F-statistics as suitable methods for the reconstruction of gene-regulatory networks from time series data. Commonly used discrete dynamic Bayesian networks perform inferior and this result can be attributed to the inevitable information loss by discretization of expression data. It is shown that short time series generated under transcription factor knock-out are optimal experiments in order to reveal the structure of gene regulatory networks. Relative to the level of observational noise, we give estimates for the required amount of gene expression data in order to accurately reconstruct gene-regulatory networks. The benefit of using of prior knowledge within a Bayesian learning framework is found to be limited to conditions of small gene expression data size. Unobserved processes, like protein-protein interactions, induce dependencies between gene expression levels similar to direct transcriptional regulation. We show that these dependencies cannot be distinguished from transcription factor mediated gene regulation on the basis of gene expression data alone. CONCLUSION: Currently available data size and data quality make the reconstruction of gene networks from gene expression data a challenge. In this study, we identify an optimal type of experiment, requirements on the gene expression data quality and size as well as appropriate reconstruction methods in order to reverse engineer gene regulatory networks from time series data. PMID- 17408502 TI - Bioinformatics strategies for lipidomics analysis: characterization of obesity related hepatic steatosis. AB - BACKGROUND: Lipids are an important and highly diverse class of molecules having structural, energy storage and signaling roles. Modern analytical technologies afford screening of many lipid molecular species in parallel. One of the biggest challenges of lipidomics is elucidation of important pathobiological phenomena from the integration of the large amounts of new data becoming available. RESULTS: We present computational and informatics approaches to study lipid molecular profiles in the context of known metabolic pathways and established pathophysiological responses, utilizing information obtained from modern analytical technologies. In order to facilitate identification of lipids, we compute the scaffold of theoretically possible lipids based on known lipid building blocks such as polar head groups and fatty acids. Each compound entry is linked to the available information on lipid pathways and contains the information that can be utilized for its automated identification from high throughput UPLC/MS-based lipidomics experiments. The utility of our approach is demonstrated by its application to the lipidomic characterization of the fatty liver of the genetically obese insulin resistant ob/ob mouse model. We investigate the changes of correlation structure of the lipidome using multivariate analysis, as well as reconstruct the pathways for specific molecular species of interest using available lipidomic and gene expression data. CONCLUSION: The methodology presented herein facilitates identification and interpretation of high-throughput lipidomics data. In the context of the ob/ob mouse liver profiling, we have identified the parallel associations between the elevated triacylglycerol levels and the ceramides, as well as the putative activated ceramide-synthesis pathways. PMID- 17408503 TI - Systems biology by the rules: hybrid intelligent systems for pathway modeling and discovery. AB - BACKGROUND: Expert knowledge in journal articles is an important source of data for reconstructing biological pathways and creating new hypotheses. An important need for medical research is to integrate this data with high throughput sources to build useful models that span several scales. Researchers traditionally use mental models of pathways to integrate information and development new hypotheses. Unfortunately, the amount of information is often overwhelming and these are inadequate for predicting the dynamic response of complex pathways. Hierarchical computational models that allow exploration of semi-quantitative dynamics are useful systems biology tools for theoreticians, experimentalists and clinicians and may provide a means for cross-communication. RESULTS: A novel approach for biological pathway modeling based on hybrid intelligent systems or soft computing technologies is presented here. Intelligent hybrid systems, which refers to several related computing methods such as fuzzy logic, neural nets, genetic algorithms, and statistical analysis, has become ubiquitous in engineering applications for complex control system modeling and design. Biological pathways may be considered to be complex control systems, which medicine tries to manipulate to achieve desired results. Thus, hybrid intelligent systems may provide a useful tool for modeling biological system dynamics and computational exploration of new drug targets. A new modeling approach based on these methods is presented in the context of hedgehog regulation of the cell cycle in granule cells. Code and input files can be found at the Bionet website: www.chip.ord/~wbosl/Software/Bionet. CONCLUSION: This paper presents the algorithmic methods needed for modeling complicated biochemical dynamics using rule-based models to represent expert knowledge in the context of cell cycle regulation and tumor growth. A notable feature of this modeling approach is that it allows biologists to build complex models from their knowledge base without the need to translate that knowledge into mathematical form. Dynamics on several levels, from molecular pathways to tissue growth, are seamlessly integrated. A number of common network motifs are examined and used to build a model of hedgehog regulation of the cell cycle in cerebellar neurons, which is believed to play a key role in the etiology of medulloblastoma, a devastating childhood brain cancer. PMID- 17408504 TI - Dynamics of in silico leukocyte rolling, activation, and adhesion. AB - BACKGROUND: We present a multilevel, agent based, in silico model that represents the dynamics of rolling, activation, and adhesion of individual leukocytes in vitro. Object-oriented software components were designed, verified, plugged together, and then operated in ways that represent the molecular and cellular mechanisms believed responsible for leukocyte rolling and adhesion. The result is an in silico analogue of an experimental in vitro system. The experimentally measured, phenotypic attributes of the analogue were compared and contrasted to those of leukocytes in vitro from three different experimental conditions. RESULTS: The individual in silico dynamics of "rolling" on simulated P-selectin, and separately on simulated VCAM-1, were an acceptable match to individual in vitro distance-time and velocity-time measurements. The analogues are also able to represent the transition from rolling to adhesion on P-selectin and VCAM-1 in the presence of GRO-alpha chemokine. The individual in silico and in vitro behavioral similarities translated successfully to population level measures. These behavioral similarities were enabled in part by subdividing the functionality of the analogue's surface into 600 independent, "cell"-controlled, equally capable modules of comparable functionality. CONCLUSION: The overlap in phenotypic attributes of our analogue with those of leukocytes in vitro confirm the considerable potential of our model for studying the key events that determine the behavioral outcome of individual leukocytes during rolling, activation, and adhesion. Our results provide an important foundation and framework for future in silico research into plausible causal links between well documented, subcellular molecular level events and the variety of systemic phenotypic attributes that distinguish normal leukocyte adhesion from abnormal disease-associated adhesion. PMID- 17408505 TI - Evolutionary conservation and over-representation of functionally enriched network patterns in the yeast regulatory network. AB - BACKGROUND: Localized network patterns are assumed to represent an optimal design principle in different biological networks. A widely used method for identifying functional components in biological networks is looking for network motifs - over represented network patterns. A number of recent studies have undermined the claim that these over-represented patterns are indicative of optimal design principles and question whether localized network patterns are indeed of functional significance. This paper examines the functional significance of regulatory network patterns via their biological annotation and evolutionary conservation. RESULTS: We enumerate all 3-node network patterns in the regulatory network of the yeast S. cerevisiae and examine the biological GO annotation and evolutionary conservation of their constituent genes. Specific 3-node patterns are found to be functionally enriched in different exogenous cellular conditions and thus may represent significant functional components. These functionally enriched patterns are composed mainly of recently evolved genes suggesting that there is no evolutionary pressure acting to preserve such functionally enriched patterns. No correlation is found between over-representation of network patterns and functional enrichment. CONCLUSION: The findings of functional enrichment support the view that network patterns constitute an important design principle in regulatory networks. However, the wildly used method of over-representation for detecting motifs is not suitable for identifying functionally enriched patterns. PMID- 17408506 TI - Predicting the connectivity of primate cortical networks from topological and spatial node properties. AB - BACKGROUND: The organization of the connectivity between mammalian cortical areas has become a major subject of study, because of its important role in scaffolding the macroscopic aspects of animal behavior and intelligence. In this study we present a computational reconstruction approach to the problem of network organization, by considering the topological and spatial features of each area in the primate cerebral cortex as subsidy for the reconstruction of the global cortical network connectivity. Starting with all areas being disconnected, pairs of areas with similar sets of features are linked together, in an attempt to recover the original network structure. RESULTS: Inferring primate cortical connectivity from the properties of the nodes, remarkably good reconstructions of the global network organization could be obtained, with the topological features allowing slightly superior accuracy to the spatial ones. Analogous reconstruction attempts for the C. elegans neuronal network resulted in substantially poorer recovery, indicating that cortical area interconnections are relatively stronger related to the considered topological and spatial properties than neuronal projections in the nematode. CONCLUSION: The close relationship between area based features and global connectivity may hint on developmental rules and constraints for cortical networks. Particularly, differences between the predictions from topological and spatial properties, together with the poorer recovery resulting from spatial properties, indicate that the organization of cortical networks is not entirely determined by spatial constraints. PMID- 17408507 TI - An in silico model of the ubiquitin-proteasome system that incorporates normal homeostasis and age-related decline. AB - BACKGROUND: The ubiquitin-proteasome system is responsible for homeostatic degradation of intact protein substrates as well as the elimination of damaged or misfolded proteins that might otherwise aggregate. During ageing there is a decline in proteasome activity and an increase in aggregated proteins. Many neurodegenerative diseases are characterised by the presence of distinctive ubiquitin-positive inclusion bodies in affected regions of the brain. These inclusions consist of insoluble, unfolded, ubiquitinated polypeptides that fail to be targeted and degraded by the proteasome. We are using a systems biology approach to try and determine the primary event in the decline in proteolytic capacity with age and whether there is in fact a vicious cycle of inhibition, with accumulating aggregates further inhibiting proteolysis, prompting accumulation of aggregates and so on. A stochastic model of the ubiquitin proteasome system has been developed using the Systems Biology Mark-up Language (SBML). Simulations are carried out on the BASIS (Biology of Ageing e-Science Integration and Simulation) system and the model output is compared to experimental data wherein levels of ubiquitin and ubiquitinated substrates are monitored in cultured cells under various conditions. The model can be used to predict the effects of different experimental procedures such as inhibition of the proteasome or shutting down the enzyme cascade responsible for ubiquitin conjugation. RESULTS: The model output shows good agreement with experimental data under a number of different conditions. However, our model predicts that monomeric ubiquitin pools are always depleted under conditions of proteasome inhibition, whereas experimental data show that monomeric pools were depleted in IMR-90 cells but not in ts20 cells, suggesting that cell lines vary in their ability to replenish ubiquitin pools and there is the need to incorporate ubiquitin turnover into the model. Sensitivity analysis of the model revealed which parameters have an important effect on protein turnover and aggregation kinetics. CONCLUSION: We have developed a model of the ubiquitin-proteasome system using an iterative approach of model building and validation against experimental data. Using SBML to encode the model ensures that it can be easily modified and extended as more data become available. Important aspects to be included in subsequent models are details of ubiquitin turnover, models of autophagy, the inclusion of a pool of short-lived proteins and further details of the aggregation process. PMID- 17408508 TI - Effect of carbon source perturbations on transcriptional regulation of metabolic fluxes in Saccharomyces cerevisiae. AB - BACKGROUND: Control effective flux (CEF) of a reaction is the weighted sum of all fluxes through that reaction, derived from elementary flux modes (EFM) of a metabolic network. Change in CEFs under different environmental conditions has earlier been proven to be correlated with the corresponding changes in the transcriptome. Here we use this to investigate the degree of transcriptional regulation of fluxes in the metabolism of Saccharomyces cerevisiae. We do this by quantifying correlations between changes in CEFs and changes in transcript levels for shifts in carbon source, i.e. between the fermentative carbon source glucose and nonfermentative carbon sources like ethanol, acetate, and lactate. The CEF analysis is based on a simple stoichiometric model that includes reactions of the central carbon metabolism and the amino acid metabolism. RESULTS: The effect of the carbon shift on the metabolic fluxes was investigated for both batch and chemostat cultures. For growth on glucose in batch (respiro-fermentative) cultures, EFMs with no by-product formation were removed from the analysis of the CEFs, whereas those including any by-products (ethanol, glycerol, acetate, succinate) were omitted in the analysis of growth on glucose in chemostat (respiratory) cultures. This resulted in improved correlations between CEF changes and transcript levels. A regression correlation coefficient of 0.60 was obtained between CEF changes and gene expression changes in the central carbon metabolism for the analysis of 5 different perturbations. Out of 45 data points there were no more than 6 data points deviating from the correlation. Additionally, up- or down-regulation of at least 75% of the genes were in qualitative agreement with the CEF changes for all perturbations studied. CONCLUSION: The analysis indicates that changes in carbon source are associated with a high degree of hierarchical regulation of metabolic fluxes in the central carbon metabolism as the change in fluxes are correlating directly with the change in transcript levels of genes encoding their corresponding enzymes. For amino acid biosynthesis there was, however, not found to exist a similar correlation, and this may point to either post-transcriptional and/or metabolic regulation, or be due to the absence of a direct perturbation on the amino acid pathways in these experiments. PMID- 17408509 TI - Structural and functional analysis of cellular networks with CellNetAnalyzer. AB - BACKGROUND: Mathematical modelling of cellular networks is an integral part of Systems Biology and requires appropriate software tools. An important class of methods in Systems Biology deals with structural or topological (parameter-free) analysis of cellular networks. So far, software tools providing such methods for both mass-flow (metabolic) as well as signal-flow (signalling and regulatory) networks are lacking. RESULTS: Herein we introduce CellNetAnalyzer, a toolbox for MATLAB facilitating, in an interactive and visual manner, a comprehensive structural analysis of metabolic, signalling and regulatory networks. The particular strengths of CellNetAnalyzer are methods for functional network analysis, i.e. for characterising functional states, for detecting functional dependencies, for identifying intervention strategies, or for giving qualitative predictions on the effects of perturbations. CellNetAnalyzer extends its predecessor FluxAnalyzer (originally developed for metabolic network and pathway analysis) by a new modelling framework for examining signal-flow networks. Two of the novel methods implemented in CellNetAnalyzer are discussed in more detail regarding algorithmic issues and applications: the computation and analysis (i) of shortest positive and shortest negative paths and circuits in interaction graphs and (ii) of minimal intervention sets in logical networks. CONCLUSION: CellNetAnalyzer provides a single suite to perform structural and qualitative analysis of both mass-flow- and signal-flow-based cellular networks in a user friendly environment. It provides a large toolbox with various, partially unique, functions and algorithms for functional network analysis. CellNetAnalyzer is freely available for academic use. PMID- 17408510 TI - Accurate, precise modeling of cell proliferation kinetics from time-lapse imaging and automated image analysis of agar yeast culture arrays. AB - BACKGROUND: Genome-wide mutant strain collections have increased demand for high throughput cellular phenotyping (HTCP). For example, investigators use HTCP to investigate interactions between gene deletion mutations and additional chemical or genetic perturbations by assessing differences in cell proliferation among the collection of 5000 S. cerevisiae gene deletion strains. Such studies have thus far been predominantly qualitative, using agar cell arrays to subjectively score growth differences. Quantitative systems level analysis of gene interactions would be enabled by more precise HTCP methods, such as kinetic analysis of cell proliferation in liquid culture by optical density. However, requirements for processing liquid cultures make them relatively cumbersome and low throughput compared to agar. To improve HTCP performance and advance capabilities for quantifying interactions, YeastXtract software was developed for automated analysis of cell array images. RESULTS: YeastXtract software was developed for kinetic growth curve analysis of spotted agar cultures. The accuracy and precision for image analysis of agar culture arrays was comparable to OD measurements of liquid cultures. Using YeastXtract, image intensity vs. biomass of spot cultures was linearly correlated over two orders of magnitude. Thus cell proliferation could be measured over about seven generations, including four to five generations of relatively constant exponential phase growth. Spot area normalization reduced the variation in measurements of total growth efficiency. A growth model, based on the logistic function, increased precision and accuracy of maximum specific rate measurements, compared to empirical methods. The logistic function model was also more robust against data sparseness, meaning that less data was required to obtain accurate, precise, quantitative growth phenotypes. CONCLUSION: Microbial cultures spotted onto agar media are widely used for genotype-phenotype analysis, however quantitative HTCP methods capable of measuring kinetic growth rates have not been available previously. YeastXtract provides objective, automated, quantitative, image analysis of agar cell culture arrays. Fitting the resulting data to a logistic equation-based growth model yields robust, accurate growth rate information. These methods allow the incorporation of imaging and automated image analysis of cell arrays, grown on solid agar media, into HTCP-driven experimental approaches, such as global, quantitative analysis of gene interaction networks. PMID- 17408511 TI - Qualitative networks: a symbolic approach to analyze biological signaling networks. AB - BACKGROUND: A central goal of Systems Biology is to model and analyze biological signaling pathways that interact with one another to form complex networks. Here we introduce Qualitative networks, an extension of Boolean networks. With this framework, we use formal verification methods to check whether a model is consistent with the laboratory experimental observations on which it is based. If the model does not conform to the data, we suggest a revised model and the new hypotheses are tested in-silico. RESULTS: We consider networks in which elements range over a small finite domain allowing more flexibility than Boolean values, and add target functions that allow to model a rich set of behaviors. We propose a symbolic algorithm for analyzing the steady state of these networks, allowing us to scale up to a system consisting of 144 elements and state spaces of approximately 10(86) states. We illustrate the usefulness of this approach through a model of the interaction between the Notch and the Wnt signaling pathways in mammalian skin, and its extensive analysis. CONCLUSION: We introduce an approach for constructing computational models of biological systems that extends the framework of Boolean networks and uses formal verification methods for the analysis of the model. This approach can scale to multicellular models of complex pathways, and is therefore a useful tool for the analysis of complex biological systems. The hypotheses formulated during in-silico testing suggest new avenues to explore experimentally. Hence, this approach has the potential to efficiently complement experimental studies in biology. PMID- 17408512 TI - METANNOGEN: compiling features of biochemical reactions needed for the reconstruction of metabolic networks. AB - BACKGROUND: One central goal of computational systems biology is the mathematical modelling of complex metabolic reaction networks. The first and most time consuming step in the development of such models consists in the stoichiometric reconstruction of the network, i. e. compilation of all metabolites, reactions and transport processes relevant to the considered network and their assignment to the various cellular compartments. Therefore an information system is required to collect and manage data from different databases and scientific literature in order to generate a metabolic network of biochemical reactions that can be subjected to further computational analyses. RESULTS: The computer program METANNOGEN facilitates the reconstruction of metabolic networks. It uses the well known database of biochemical reactions KEGG of biochemical reactions as primary information source from which biochemical reactions relevant to the considered network can be selected, edited and stored in a separate, user-defined database. Reactions not contained in KEGG can be entered manually into the system. To aid the decision whether or not a reaction selected from KEGG belongs to the considered network METANNOGEN contains information of SWISSPROT and ENSEMBL and provides Web links to a number of important information sources like METACYC, BRENDA, NIST, and REACTOME. If a reaction is reported to occur in more than one cellular compartment, a corresponding number of reactions is generated each referring to one specific compartment. Transport processes of metabolites are entered like chemical reactions where reactants and products have different compartment attributes. The list of compartmentalized biochemical reactions and membrane transport processes compiled by means of METANNOGEN can be exported as an SBML file for further computational analysis. METANNOGEN is highly customizable with respect to the content of the SBML output file, additional data fields, the graphical input form, highlighting of project specific search terms and dynamically generated Web-links. CONCLUSION: METANNOGEN is a flexible tool to manage information for the design of metabolic networks. The program requires Java Runtime Environment 1.4 or higher and about 100 MB of free RAM and about 200 MB of free HD space. It does not require installation and can be directly Java webstarted from http://3d-alignment.eu/metannogen/. PMID- 17408513 TI - Stochastic synchronization of genetic oscillator networks. AB - BACKGROUND: The study of synchronization among genetic oscillators is essential for the understanding of the rhythmic phenomena of living organisms at both molecular and cellular levels. Genetic networks are intrinsically noisy due to natural random intra- and inter-cellular fluctuations. Therefore, it is important to study the effects of noise perturbation on the synchronous dynamics of genetic oscillators. From the synthetic biology viewpoint, it is also important to implement biological systems that minimizing the negative influence of the perturbations. RESULTS: In this paper, based on systems biology approach, we provide a general theoretical result on the synchronization of genetic oscillators with stochastic perturbations. By exploiting the specific properties of many genetic oscillator models, we provide an easy-verified sufficient condition for the stochastic synchronization of coupled genetic oscillators, based on the Lur'e system approach in control theory. A design principle for minimizing the influence of noise is also presented. To demonstrate the effectiveness of our theoretical results, a population of coupled repressillators is adopted as a numerical example. CONCLUSION: In summary, we present an efficient theoretical method for analyzing the synchronization of genetic oscillator networks, which is helpful for understanding and testing the synchronization phenomena in biological organisms. Besides, the results are actually applicable to general oscillator networks. PMID- 17408514 TI - Synthetic tetracycline-inducible regulatory networks: computer-aided design of dynamic phenotypes. AB - BACKGROUND: Tightly regulated gene networks, precisely controlling the expression of protein molecules, have received considerable interest by the biomedical community due to their promising applications. Among the most well studied inducible transcription systems are the tetracycline regulatory expression systems based on the tetracycline resistance operon of Escherichia coli, Tet-Off (tTA) and Tet-On (rtTA). Despite their initial success and improved designs, limitations still persist, such as low inducer sensitivity. Instead of looking at these networks statically, and simply changing or mutating the promoter and operator regions with trial and error, a systematic investigation of the dynamic behavior of the network can result in rational design of regulatory gene expression systems. Sophisticated algorithms can accurately capture the dynamical behavior of gene networks. With computer aided design, we aim to improve the synthesis of regulatory networks and propose new designs that enable tighter control of expression. RESULTS: In this paper we engineer novel networks by recombining existing genes or part of genes. We synthesize four novel regulatory networks based on the Tet-Off and Tet-On systems. We model all the known individual biomolecular interactions involved in transcription, translation, regulation and induction. With multiple time-scale stochastic-discrete and stochastic-continuous models we accurately capture the transient and steady state dynamics of these networks. Important biomolecular interactions are identified and the strength of the interactions engineered to satisfy design criteria. A set of clear design rules is developed and appropriate mutants of regulatory proteins and operator sites are proposed. CONCLUSION: The complexity of biomolecular interactions is accurately captured through computer simulations. Computer simulations allow us to look into the molecular level, portray the dynamic behavior of gene regulatory networks and rationally engineer novel ones with useful applications. We are able to propose, test and accept or reject design principles for each network. Guided by simulations, we develop a set of design principles for novel tetracycline-inducible networks. PMID- 17408515 TI - Identification of functional modules using network topology and high-throughput data. AB - BACKGROUND: With the advent of systems biology, biological knowledge is often represented today by networks. These include regulatory and metabolic networks, protein-protein interaction networks, and many others. At the same time, high throughput genomics and proteomics techniques generate very large data sets, which require sophisticated computational analysis. Usually, separate and different analysis methodologies are applied to each of the two data types. An integrated investigation of network and high-throughput information together can improve the quality of the analysis by accounting simultaneously for topological network properties alongside intrinsic features of the high-throughput data. RESULTS: We describe a novel algorithmic framework for this challenge. We first transform the high-throughput data into similarity values, (e.g., by computing pairwise similarity of gene expression patterns from microarray data). Then, given a network of genes or proteins and similarity values between some of them, we seek connected sub-networks (or modules) that manifest high similarity. We develop algorithms for this problem and evaluate their performance on the osmotic shock response network in S. cerevisiae and on the human cell cycle network. We demonstrate that focused, biologically meaningful and relevant functional modules are obtained. In comparison with extant algorithms, our approach has higher sensitivity and higher specificity. CONCLUSION: We have demonstrated that our method can accurately identify functional modules. Hence, it carries the promise to be highly useful in analysis of high throughput data. PMID- 17408516 TI - Decision tree modeling predicts effects of inhibiting contractility signaling on cell motility. AB - BACKGROUND: Computational models of cell signaling networks typically are aimed at capturing dynamics of molecular components to derive quantitative insights from prior experimental data, and to make predictions concerning altered dynamics under different conditions. However, signaling network models have rarely been used to predict how cell phenotypic behaviors result from the integrated operation of these networks. We recently developed a decision tree model for how EGF-induced fibroblast cell motility across two-dimensional fibronectin-coated surfaces depends on the integrated activation status of five key signaling nodes, including a proximal regulator of transcellular contractile force generation, MLC (myosin light chain) [Hautaniemi et al, Bioinformatics 21: 2027 {2005}], but we have not previously attempted predictions of new experimental effects from this model. RESULTS: In this new work, we construct an improved decision tree model for the combined influence of EGF and fibronectin on fibroblast cell migration based on a wider spectrum of experimental protein signaling and cell motility measurements, and directly test a significant and non-intuitive a priori prediction for the outcome of a targeted molecular intervention into the signaling network: that partially reducing activation of MLC would increase cell motility on moderately adhesive surfaces. This prediction was indeed confirmed experimentally: partial inhibition of the activating MLC kinase (MLCK) upstream using the pharmacologic agent ML-7 resulted in increased motility of NR6 fibroblasts. We further extended this exciting finding by showing that partial reduction of MLC activation similarly enhanced the transmigration of the human breast carcinoma cell line MDA-213 through a Matrigel barrier. CONCLUSION: These findings specifically highlight a central regulatory role for transcellular contractility in governing cell motility, while at the same time demonstrating the value of a decision tree approach to a systems "signal-response" model in discerning non-intuitive behavior arising from integrated operation a cell signaling network. PMID- 17408517 TI - Effects of three different conjugated linoleic acid preparations on insulin signalling, fat oxidation and mitochondrial function in rats fed a high-fat diet. AB - To investigate the effects of three different conjugated linoleic acid (CLA) preparations containing different ratios of CLA isomers on insulin signalling, fatty acid oxidation and mitochondrial function, Sprague-Dawley rats were fed a high-fat diet either unsupplemented or supplemented with one of three CLA preparations at 1 % of the diet for 8 weeks. The first CLA preparation contained approximately 30 % cis-9, trans-11 (c9, t11)-CLA isomer and 40 % trans-10, cis-12 (t10, c12)-CLA isomer (CLA-mix). The other two preparations were an 80:20 mix (c9, t11-CLA-mix) or a 10:90 mix of two CLA isomers (t10, c12-CLA-mix). Insulin resistance was decreased in all three supplemented groups based on the results of homeostasis model assessment and the revised quantitative insulin-sensitivity check index. The phosphorylation of insulin receptor substrate-1 on serine decreased in the livers of all three supplemented groups, while subsequent Akt phosphorylation increased only in the t10, c12-CLA-mix group. Both the c9, t11 CLA-mix and the t10, c12-CLA-mix increased the expression of hepatic adiponectin receptors R1 and 2, which are thought to enhance insulin sensitivity and fat oxidation. The c9, t11-CLA-mix increased protein and mRNA levels of PPAR alpha, acyl-CoA oxidase and uncoupling protein, which are involved in fatty acid oxidation and energy dissipation. The c9, t11-CLA-mix enhanced mitochondrial function and protection against oxidative stress by increasing the activities of cytochrome c oxidase, manganese-superoxide dismutase, glutathione peroxidase, and glutathione reductase and the level of GSH. In conclusion, all three CLA preparations reduced insulin resistance. Among them, the c9, t11-CLA-mix was the most effective based on the parameters reflecting insulin resistance and fat oxidation, and mitochondrial antioxidative enzyme activity in the liver. PMID- 17408518 TI - Dietary intake and a food-frequency instrument to evaluate a nutrition intervention for the Apache in Arizona. AB - OBJECTIVE: To demonstrate how dietary data collected from 24-hour dietary recalls were incorporated into the development of a quantitative food-frequency questionnaire (QFFQ) for the Apache in Arizona. DESIGN: A cross-sectional study was first conducted using 24-hour dietary recalls to identify foods for inclusion on a QFFQ that would be used to evaluate a nutrition intervention. SETTING: The White Mountain and San Carlos Apache reservations in East-central Arizona. SUBJECTS: The 24-hour dietary recalls were collected from a random sample of 53 adults (34 women and 19 men). RESULTS: A QFFQ was developed that included all foods reported by two or more respondents, plus traditional and seasonal foods. Portion size was assessed using familiar household units and culturally appropriate food models. The final instrument contains 155 foods. Frequency of consumption is assessed using eight categories ranking from 'never or less than once a month' to '2 or more times a day'. Nutrient intakes and the five major food sources of energy, fat and sugar are presented. CONCLUSION: The QFFQ developed is complete and up-to-date for assessing usual food and nutrient intake for the Apache in Arizona. The instrument will be used to evaluate a food store based nutrition intervention to reduce risk of chronic diseases. PMID- 17408519 TI - In appreciation of Sir Philip Randle: the glucose-fatty acid cycle. AB - The coordinated regulation of metabolic fuel selection is crucial to energy homeostasis. Philip Randle and his colleagues developed the fundamental concept of interplay between carbohydrate and lipid fuels in relation to the requirement for energy utilisation and storage. Their insight has fashioned current understanding of the regulation of metabolism in health and disease, as well as providing a springboard for research into the roles of lipid derivatives in insulin resistance and, at the transcriptional level, lipid-regulated nuclear hormone receptors. PMID- 17408520 TI - Arginine supplementation for neonatal necrotizing enterocolitis: are we ready? PMID- 17408521 TI - Hypocholesterolaemic effects of soya proteins: results of recent studies are predictable from the anderson meta-analysis data. AB - In 1995, Anderson et al. published a meta-analysis, derived from most of the clinical studies on soya proteins given to individuals with varying levels of cholesterolaemia that had been reported up to that time. The meta-analysis clearly indicated that cholesterolaemias were generally reduced by diets with soya given as a partial or total substitution of animal proteins, with final mean total and LDL-cholesterol reductions of 23.2 mg/dl and 21.7 mg/dl, respectively. These findings were recently strongly criticised, based on the evaluation of later studies, frequently involving individuals with normal or moderately elevated cholesterolaemias. In the present paper, these more recent studies were re-evaluated using a 'nomogram' prepared on the basis of the quartiles of initial cholesterol concentrations in the Anderson meta-analysis and their corresponding CI for net cholesterol change. The five studies belonging to the first quartile and thirteen out of the fourteen belonging to the second quartile gave results perfectly in line with the nomogram. Out of the fourteen studies belonging to the third quartile, ten agreed with the nomogram and two gave lower cholesterol reductions, whereas two gave higher reductions. Unfortunately, none of the recent studies belonged to the fourth quartile as treatment with statins or other lipid lowering drugs is now mandatory in the presence of very high cholesterol levels. The re-evaluation thus shows that the thirty-three studies published in the past 10 years are in agreement with the Anderson meta-analysis and confirm its validity. PMID- 17408522 TI - N-3 fatty acids, cancer and cachexia: a systematic review of the literature. AB - Use of n-3 fatty acids (FA) has been reported to be beneficial for cancer patients. We performed a systematic review of the literature in order to issue recommendations on the clinical use of n-3 FA in the cancer setting. A systematic search was performed in MEDLINE, EMBASE, Cochrane and Healthstar databases. We selected clinical trials or prospective observational studies including patients with cancer and life expectancy >2 months, in which enteral supplements with n-3 FA were administered. Parameters evaluated individually were clinical (nutritional status, tolerance, survival and hospital stays), biochemical (inflammatory mediators), and functional (functional status, appetite and quality of life (QoL)). Seventeen studies met the inclusion criteria; eight were of high quality. The panel of experts established the following evidence: (1) oral supplements with n-3 FA benefit patients with advanced cancer and weight loss, and are indicated in tumours of the upper digestive tract and pancreas; (2) the advantages observed were: increased weight and appetite, improved QoL, and reduced post-surgical morbidity; (3) there is no defined pattern for combining different n-3 FA, and it is recommended to administer > 1.5 g/day; and (4) better tolerance is obtained administering low-fat formulas for a period of at least 8 weeks. All the evidences were grade B but for 'length of treatment' and 'advantage of survival' it was grade C. Our findings suggest that administration of n-3 FA (EPA and DHA) in doses of at least 1.5 g/day for a prolonged period of time to patients with advanced cancer is associated with an improvement in clinical, biological and QoL parameters. PMID- 17408523 TI - Associations between dietary added sugar intake and micronutrient intake: a systematic review. AB - There is increasing concern that high intakes of added sugars might compromise intakes of micronutrients. The objectives of this systematic review were (1) to determine whether dietary added sugar intake was associated with micronutrient intakes, and if so, whether there was evidence of micronutrient dilution as a result of higher dietary added sugar intake and (2) if micronutrient dilution was present, to determine whether there was sufficiently robust evidence to support a threshold effect above which there was a significant decline in micronutrient intake or status relative to the recommended intakes. A systematic computerised literature search was undertaken, limited to studies written in English published from 1980 onwards and further studies identified through hand searching papers. Fifteen studies that assessed associations between intakes of added sugars or non milk extrinsic sugars and micronutrients were included. Overall, there are insufficient data and inconsistency between studies in relationships between added sugars and micronutrient intakes, with no clear evidence of micronutrient dilution or a threshold for a quantitative amount of added sugar intake for any of the micronutrients investigated. The current evidence base is considerably constrained by methodological issues. Further research is required to determine which food products high in added sugars might adversely affect micronutrient intakes by displacing other food items from the diet. Analyses should take into account the magnitude of any observed associations to determine their true biological significance. PMID- 17408524 TI - Acute effects of tea on fasting and non-fasting plasma total homocysteine concentrations in human subjects. AB - Plasma total homocysteine concentrations (tHcy) are a putative risk factor for CVD. Tea is a rich dietary source of polyphenols and caffeine, both of which may raise tHcy. However, it is possible that much of any effect is transitory and may be influenced by the consumption of food. Our objective was to investigate the acute effect of tea, at a dose representative of ordinary population intakes, on tHcy and to determine whether consumption of a meal influences the magnitude of any effect. Measurements of tHcy were performed in twenty participants at baseline and 3.5 h after drinking three cups of black tea or hot water (consumed at time 0, 1.5 and 3 h) with and without a meal: a total of four treatments administered in random order. Drinking tea resulted in an acute increase in tHcy (0-30 (95 % CI 0.04, 0.56) micromol/l, P=0.022). The meal resulted in an acute decrease in tHcy (-0.42 (95 % CI -0.68, -0.16) micromol/l, P=0.002). There was no interaction between tea and meal on tHcy (P=0.40); that is, the effect of tea on tHcy was not different in the fasting and non-fasting state. Our results suggest that drinking black tea can cause a small acute increase in tHcy and that this effect is not enhanced in the non-fasting state. Given that results of population studies have generally shown a negative association between tea intake and tHcy, the significance of these findings to CVD risk remains uncertain. PMID- 17408525 TI - The minor allele of the PPARgamma2 pro12Ala polymorphism is associated with lower postprandial TAG and insulin levels in non-obese healthy men. AB - The PPARgamma Pro12Ala polymorphism has been associated in several studies with a decreased risk of obesity, type 2 diabetes and insulin resistance. Weak hints are available about the influence of PPARgamma Pro12Ala on postprandial metabolism. In 708 men, aged 45 to 65 years the PPARgamma2 Pro12Ala genotypes were determined and postprandial TAG, insulin, glucose and NEFA after a standardized mixed fat meal and insulin and glucose after a glucose load (oral glucose tolerance test; OGTT) were assessed. Using the total sample, we did not find a significant impact of the genotype on the postprandial metabolism. In the subgroup with BMI <30 kg/m2, fasting and postprandial TAG and insulin levels as well as homeostasis model assessment of insulin resistance (HOMA) were significantly lower in the Ala12Ala group than in the Pro12Pro group after the mixed meal. In contrast, the groups did not differ in insulin levels and HOMA after the OGTT. To investigate if differences between a fat-containing meal and OGTT are caused by adiponectin, we examined a BMI- and age-matched subgroup. No differences were found between the genotypic groups. The effects of the PPARgamma2 polymorphism on insulin sensitivity are mediated by affluent dietary fat. We did not find evidence that adiponectin as a fatty-acid-dependent adipocyte factor is a causative factor for this phenomenon. PMID- 17408526 TI - Interactions between protein and vegetable oils in the maternal diet determine the programming of the insulin axis in the rat. AB - The available evidence suggests that metabolic control mechanisms are programmed early in life. Previous studies of pregnant rats fed low-protein diets have suggested that the vegetable oils used in the experimental diets influence the outcome. The present study investigated the offspring of female rats fed semi synthetic diets containing either 180 or 90g casein/kg with 70 g/kg (w/w) of either corn oil or soya oil during gestation. During lactation, the dams received stock diet, and the offspring were subsequently weaned onto the stock diet. The offspring of dams fed the low-protein diets were smaller at birth. At 25 weeks of age, the offspring were subjected to an oral glucose tolerance test. In the offspring of dams fed the diet containing soya oil, the area under the insulin curve was affected by the protein content of the maternal diet. There was no effect of protein on the area under the insulin curve in the offspring of dams fed the diet prepared with corn oil. There were no differences in plasma glucose concentrations. The levels of mRNA for acetyl-CoA carboxylase- in the livers of female offspring were affected by the protein and oil content of the maternal diet. The level of carnitine palmitoyl transferase mRNA was affected by the protein content of the maternal diet. The present study suggests that PUFA in the maternal diet can interact with protein metabolism to influence the development of the offspring. This may involve the higher content of alpha-linolenic acid in soya oil compared with corn oil. PMID- 17408527 TI - Undernutrition in early life and body composition of adolescent males from a birth cohort study. AB - The evidence for an association between poor nutrition in early life and subsequent obesity is inconclusive. In the present study, we investigated the associations between stunting, wasting and underweight at 2 and 4 years of age, and body composition in adolescence in male subjects studied since birth. The 1982 Pelotas Birth Cohort Study included all children born in maternity hospitals and living in the urban area of the city of Pelotas, Southern Brazil. All males born in 1982 were legally required to enlist in the army between January and April 2000. We were thus able to track 2250 subjects in 2000 (78.9% of the original cohort). Anthropometric measurements were collected in 1984 and 1986, and body composition was assessed in 2000. In the present analysis, we used as predictors the nutritional indices height-for-age, weight-for-height and weight for-age presented in six categories. Outcomes included fat, lean and body mass indices and fat:lean mass ratio, derived from anthropometric and bioimpedance measurements. ANOVA and linear regression were used in the analyses to adjust for confounding. All predictors were positively associated with fat and body mass indices. Height-for-age Z score at age 2 or 4 years was not associated with lean mass index, but all other predictors were associated. Fat:lean mass ratio was associated only with weight-for-height Z score. Our results suggest that undernutrition is not a risk factor for overweight and obesity in our population and may partially protect against fatness in adolescence. PMID- 17408528 TI - Absorption and metabolism of bioactive molecules after oral consumption of cooked edible heads of Cynara scolymus L. (cultivar Violetto di Provenza) in human subjects: a pilot study. AB - The current growing interest for natural antioxidants has led to a renewed scientific attention for artichoke, due not only to its nutritional value, but, overall, to its polyphenolic content, showing strong antioxidant properties. The major constituents of artichoke extracts are hydroxycinnamic acids such as chlorogenic acid, dicaffeoylquinic acids caffeic acid and ferulic acid, and flavonoids such as luteolin and apigenin glycosides. In vitro studies, using cultured rat hepatocytes, have shown its hepatoprotective functions and in vivo studies have shown the inhibition of cholesterol biosynthesis in human subjects. Several studies have shown the effect on animal models of artichoke extracts, while information on human bioavailability and metabolism of hydroxycinnamates derivatives is still lacking. Results showed a plasma maximum concentration of 6.4 (SD 1.8) ng/ml for chlorogenic acid after 1 h and its disappearance within 2 h (P< 0.05). Peak plasma concentrations of 19.5 (SD 6.9) ng/ml for total caffeic acid were reached within 1 h, while ferulic acid plasma concentrations showed a biphasic profile with 6.4 (SD1.5) ng/ml and 8.4 (SD4.6) ng/ml within 1 h and after 8 h respectively. We observed a significant increase of dihydrocaffeic acid and dihydroferulic acid total levels after 8 h (P<0.05). No circulating plasma levels of luteolin and apigenin were present. Our study confirms the bioavailability of metabolites of hydroxycinnamic acids after ingestion of cooked edible Cynara scolymus L. (cultivar Violetto di Provenza). PMID- 17408529 TI - Plasma concentrations of carotenoids and vitamin C are better correlated with dietary intake in normal weight than overweight and obese elderly subjects. AB - Carotenoid and vitamin C intakes, assessed by FFQ, have been positively associated with plasma concentrations in different populations. However, the influence of BMI on these associations has not been explored in detail. We explored in a cross-sectional study the relation between dietary carotenoid and vitamin C intakes, using a 135-item FFQ, with their plasma concentrations by BMI categories in 252 men and 293 women, 65 years and older. For men and women combined, significant (P < 0.05) Pearson correlations were observed between energy-adjusted dietary intakes and plasma concentrations (carotenoids adjusted for cholesterol) for: alpha-carotene 0.21, beta-carotene 0.19, lycopene 0.18, beta-cryptoxanthin 0.20 and vitamin C 0.36. Multiple linear regression analyses showed that the intake of carotenoids and vitamin C were significant predictors of their respective plasma concentration (P<0.01), and that BMI was inversely associated with plasma concentration of carotenoids (P< or =0.01) but not with plasma vitamin C. In addition, we observed significant interactions between BMI and the intakes of alpha-carotene and lutein + zeaxanthin, and to a lower extent beta-carotene, suggesting that these intakes in subjects with high BMI were not good predictors of their plasma concentration. The present data suggest that plasma carotenoids and vitamin C may be good markers of dietary intake in elderly subjects, but not so for alpha-carotene, beta-carotene and lutein + zeaxanthin in obese subjects. PMID- 17408530 TI - Black Lucques olives prevented bone loss caused by ovariectomy and talc granulomatosis in rats. AB - This study was conducted to determine whether olive fruits, rich in micronutrients, might improve bone loss in ovariectomized (OVX) rats (an experimental model of postmenopausal osteoporosis) and in OVX rats with granulomatosis inflammation (a model of senile osteoporosis). Six-month-old Wistar female rats underwent ovariectomy and were then immediately treated orally by substituting oil in the diet by 10 g/d green Lucques olives or 6 g/d black Lucques olives for each rat for 84 days. OVX rats and sham-operated controls received the same diet with oil. Three weeks before the end of the experiment, subcutaneous inflammation was provoked by injections of sterile magnesium silicate in half the animals in each group. In OVX rats, granulomatosis inflammation, characterized by a rise in inflammatory parameters such as fibrinogen, alpha1-acid glycoprotein, spleen weight and granulocyte level, and an impairment of oxidative status (as shown by a decrease in plasma antioxidant capacity, a higher rate of isoprostane excretion) elicited a bone loss in the whole femur and in the metaphyseal areas considered on their own. Whereas green olives had no effect on osteopenia, consumption of the black variety prevented bone loss in the whole femur and at cortical sites in those oestrogen-deficient animals with talc inflammation (diaphyseal bone mineral density: black olives and inflammation 0-2323 (SE 0.0026) v. ovariectomy and inflammation 0.2117 (SE 0.0030); P=0.027). This bone-sparing effect seemed to result from an improvement in the inflammatory and oxidative status. The present data show that black olives are able to prevent bone loss in an experimental model of senile osteoporosis (oestrogen-deficient rats in which a low-grade inflammation was induced by talc injection). PMID- 17408531 TI - Docosahexaenoic acid and human brain evolution: missing the forest for the trees- comments by Cunnane. PMID- 17408532 TI - Is docosahexaenoic acid a red herring for the aquatic diet?--Comments by Milligan and Bazinet. PMID- 17408533 TI - Recruitment of the serratus anterior as an accessory muscle of ventilation during graded exercise. AB - The role of the serratus anterior (SA) as an accessory muscle of ventilation and its physiologic significance under exercising conditions remains unclear. Recent investigations have utilized the measurement of SA as an analog for respiratory muscle oxygenation. The purpose of this investigation was to examine the action of the serratus anterior via surface electromyography (EMG) and near infrared spectroscopy (NIRS) during exercise while controlling for muscular effort not related to ventilation. Nine healthy volunteers (age = 24.4 +/- 0.5 years, VO2max = 3.416 +/- 0.35 l min(-1); VEpeak = 127.5 +/- 13.1 l min(-1); TVpeak = 2.844 +/- 0.226 l) completed a graded exercise test to volitional exhaustion on a cycle ergometer. The subjects' arms were folded and relaxed at the abdomen to minimize muscular effort resulting from scapular stabilization during pushing/forward flexion of the arms associated with cycle ergometry. VO2 and V were monitored breath-by-breath throughout exercise. EMG was recorded over the right SA, and a near infrared probe was placed over the left SA. No significant differences were observed throughout the graded exercise test for tissue oxygenation (StO2) (n = 6, F[1.532, 7.661] = 0.895, P > 0.05, eta2 = 0.15) or EMG (n = 9, F[1.594, 12.75] = 3.067, P > 0.05, eta2 = 0.27). Although the recruitment of the SA has been postulated to aid in ventilation in various postures and disease states, it is concluded that it shows little muscular effort in healthy subjects during upright cycling. Additional research is needed to conclude the pertinence of utilizing StO2 of the SA as an analog for respiratory muscle oxygenation. PMID- 17408534 TI - A systematic review and economic evaluation of epoetin alpha, epoetin beta and darbepoetin alpha in anaemia associated with cancer, especially that attributable to cancer treatment. AB - OBJECTIVES: To assess the effectiveness and cost-effectiveness of epoetin alpha, epoetin beta and darbepoetin alpha (referred to collectively in this report as epo) in anaemia associated with cancer, especially that attributable to cancer treatment. DATA SOURCES: Electronic databases were searched from 2000 (1996 in the case of darbepoetin alpha) to September 2004. REVIEW METHODS: Using a recently published Cochrane review as the starting point, a systematic review of recent randomised controlled trials (RCTs) comparing epo with best standard was conducted. Inclusion, quality assessment and data abstraction were undertaken in duplicate. Where possible, meta-analysis was employed. The economic assessment consisted of a systematic review of past economic evaluations, an assessment of economic models submitted by the manufacturers of the three epo agents and development of a new individual sampling model (the Birmingham epo model). RESULTS: In total 46 RCTs were included within this systematic review, 27 of which had been included in the Cochrane systematic review. All 46 trials compared epo plus supportive care for anaemia (including transfusions), with supportive care for anaemia (including transfusions), alone. Haematological response (defined as an improvement by 2 g/dl(-1)) had a relative risk of 3.4 [95% confidence interval (CI) 3.0 to 3.8, 22 RCTs] with a response rate for epo of 53%. The trial duration was most commonly 16-20 weeks. There was little statistical heterogeneity in the estimate of haematological response, and there were no important differences between the subgroups examined. Haemoglobin (Hb) change showed a weighted mean difference of 1.63 g/dl(-1) (95% CI 1.46 to 1.80) in favour of epo. Treatment with erythropoietin in patients with cancer-induced anaemia reduces the number of patients who receive a red blood cell transfusion (RBCT) by an estimated 18%. Health-related quality of life (HRQoL) data were analysed using vote counting and qualitative assessment and a positive effect was observed in favour of an improved HRQoL for patients on epo. Published information on side-effects was of poor quality. New trials provided further evidence of side-effects with epo, particularly thrombic events, but it is still unclear whether these could be accounted for by chance alone. The results of the previous Cochrane review had suggested a survival advantage for epo (HR 0.84, 95% CI 0.69 to 1.02), based on 19 RCTs. The update, based on 28 RCTs, suggests no difference (HR 1.03, 95% CI 0.88 to 1.21). Subgroup analysis suggested some explanations for this heterogeneity, but it is difficult to draw firm conclusions without access to the substantial amounts of missing or unpublished data, or more detailed results from some of the trials with heterogeneous patient populations. The conclusions are, however, broadly in line with those of a Food and Drug Administration (FDA) safety briefing, which recommended that patients with a haemoglobin above 12 g/dl(-1) should not be treated; the target rate of rise in Hb should not be too great, and further carefully conducted trials are required to determine which subgroups of patients may be harmed by the use of these products, in particular through the stimulation of tumour activity. Five published economic evaluations identified from the literature had inconsistent results, with estimates ranging from a cost per quality-adjusted life-year (QALY) under pound 10,000 through to epo being less effective and more costly than standard care. The more favourable evaluations assumed a survival advantage for epo. The three company models submitted each relied on assumed survival gains to achieve relatively low cost per QALY, from pound 13,000 to pound 28,000, but generated estimates from pound 84,000 to pound 159,000 per QALY when no survival gain was assumed. Each of these models relied on Hb levels alone driving utility, and each assumed gradual normalisation of Hb in the standard treatment arm after the end of treatment. The Birmingham epo model followed the company models in regard to the relationship between Hb levels and utility, and also assumed normalisation in the base case. With no survival gain, the incremental cost per QALY was pound 150,000, falling to pound 40,000 when the lower, more favourable, confidence interval for survival was used. CONCLUSIONS: Epo is effective in improving haematological response and reducing RBCT requirements, and appears to have a positive effect on HRQoL. The incidence of side-effects and effects on survival remains highly uncertain. However, if there is no impact on survival, it seems highly unlikely that epo would be considered a cost-effective use of healthcare resources. The main target for further research should be improving estimates of impact on survival, initially through more detailed secondary research, such as the individual patient data meta-analysis started by the Cochrane group. Further trials may be required, and have been recommended by the FDA, although many trials are in progress, completed but unreported or awaiting mature follow-up. The Birmingham epo model developed as part of this project contains new features that improve its flexibility in exploring different scenarios; further refinement and validation would therefore be of assistance. Finally, further research to resolve uncertainty about other parameters, particularly quality of life, adverse events, and the rate of normalisation, would also be beneficial. PMID- 17408535 TI - A systematic review and economic evaluation of statins for the prevention of coronary events. AB - OBJECTIVES: To evaluate the clinical effectiveness and cost-effectiveness of statins for the primary and secondary prevention of cardiovascular events in adults with, or at risk of, coronary heart disease (CHD). DATA SOURCES: Electronic databases were searched between November 2003 and April 2004. REVIEW METHODS: A review was undertaken to identify and evaluate all literature relating to the clinical and cost effectiveness of statins in the primary and secondary prevention of CHD and cardiovascular disease (CVD) in the UK. A Markov model was developed to explore the costs and health outcomes associated with a lifetime of statin treatment using a UK NHS perspective. RESULTS: Thirty-one randomised studies were identified that compared a statin with placebo or with another statin, and reported clinical outcomes. Meta-analysis of the available data from the placebo-controlled studies indicates that, in patients with, or at risk of, CVD, statin therapy is associated with a reduced relative risk of all cause mortality, cardiovascular mortality, CHD mortality and fatal myocardial infarction (MI), but not of fatal stroke. It is also associated with a reduced relative risk of morbidity [non-fatal stroke, non-fatal MI, transient ischaemic attack (TIA), unstable angina] and of coronary revascularisation. It is hardly possible, on the evidence available from the placebo-controlled trials, to differentiate between the clinical efficacy of atorvastatin, fluvastatin, pravastatin and simvastatin. However, there is some evidence from direct comparisons between statins to suggest that atorvastatin may be more effective than pravastatin in patients with symptomatic CHD. There is limited evidence for the effectiveness of statins in different subgroups. Statins are generally considered to be well tolerated and to have a good safety profile. This view is generally supported both by the evidence of the trials included in this review and by postmarketing surveillance data. Increases in creatine kinase and myopathy have been reported, but rhabdomyolysis and hepatotoxicity are rare. However, some patients may receive lipid-lowering therapy for as long as 50 years, and long term safety over such a timespan remains unknown. In secondary prevention of CHD, the incremental cost-effectiveness ratios (ICERs) increase with age varying between pound 10,000 and pound 17,000 per quality adjusted life year (QALY) for ages 45 and 85 respectively. Sensitivity analyses show these results are robust. In primary prevention of CHD there is substantial variation in ICERs by age and risk. The average ICERs weighted by risk range from pound 20,000 to pound 27,500 for men and from pound 21,000 to pound 57,000 for women. The results are sensitive to the cost of statins, discount rates and the modelling time frame. In the CVD analyses, which take into account the benefits of statins on reductions in stroke and TIA events, the average ICER weighted by risk level remains below pound 20,000 at CHD risk levels down to 0.5%. Limitations of the analyses include the requirement to extrapolate well beyond the timeframe of the trial period, and to extrapolate effectiveness results from higher risk primary prevention populations to the treatment of populations at much lower risk. Consequently, the results for the lower age bands and lower risks are subject to greater uncertainty and need to be treated with caution. CONCLUSIONS: There is evidence to suggest that statin therapy is associated with a statistically significant reduction in the risk of primary and secondary cardiovascular events. As the confidence intervals for each outcome in each prevention category overlap, it is not possible to differentiate, in terms of relative risk, between the effectiveness of statins in primary and secondary prevention. However, the absolute risk of CHD death/non-fatal MI is higher, and the number needed to treat to avoid such an event is consequently lower, in secondary than in primary prevention. The generalisability of these results is limited by the exclusion, in some studies, of patients who were hypersensitive to, intolerant of, or known to be unresponsive to, statins, or who were not adequately compliant with study medication during a placebo run-in phase. Consequently, the treatment effect may be reduced when statins are used in an unselected population. The results of the economic modelling show that statin therapy in secondary prevention is likely to be considered cost-effective. In primary prevention, the cost-effectiveness ratios are dependent on the level of CHD risk and age, but the results for the CVD analyses offer support for the more aggressive treatment recommendation issued by recent guidelines in UK. Evidence on clinical endpoints for rosuvastatin is awaited from on-going trials. The potential targeting of statins at low-risk populations is however associated with major uncertainties, particularly the likely uptake and long-term compliance to lifelong medication by asymptomatic younger patients. The targeting, assessment and monitoring of low risk patients in primary care would be a major resource implication for the NHS. These areas require further research. PMID- 17408536 TI - [Predictors of nosocomial infection in acute stroke. Relation with morbimortality and outcome]. AB - BACKGROUND AND OBJECTIVE: Stroke is a very important cause of mortality and disability. This study has the objective of identifying predictor factors and the clinical consequences of nosocomial infection in acute stroke. PATIENTS AND METHOD: We prospectively identified a consecutive cohort of patients who were admitted after an acute stroke. We used predefined diagnostic criteria by the World Health Organization and Sociedad Espanola de Neurologia for stroke, and by Centers for Disease Control and Sociedad Espanola de Enfermedades Infecciosas y Microbiologia Clinica for infection. RESULTS: 258 patients with acute stroke were included. 102 (39.5%) had at least one nosocomial infection -45.5% women; age (standard deviation) 78.2 (9.7) years-. The mean hospital stay was 14.9 days (8.4) in infection patients and 8.4 days (5.6) in no infection patients (p < 0.001). 31 patients died and 22 (71%) had at least one cause of infection. Using logistic regression analysis, the dysphagia (odds ratio [OR] = 12.7; 95% confidence interval [CI], 5.3-30.1; p < 0.001) is the strongest and independent predictor of nosocomial infection. Others factors are crural motor affectation (OR = 4.5; 95% CI, 1.7-12.3; p = 0.003), urinary incontinence (OR = 2.9; 95% CI, 1.3-6.4; p = 0.009) and diabetes mellitus (OR = 2.3; 95% CI, 1.1-4.7; p = 0.03). Baseline imbalance National Institutes of Health Stroke Scale (NIHSS) > 20 during the admission (OR = 17.3; 95% CI, 5.1-59.5; p < 0.001), mass effect diagnosticated on computerized axial tomography (OR = 4.4; 95% CI, 1.4-14; p = 0.012), poor neurological outcome during the first day (OR = 11.6; 95% CI, 3.6 37.2; p < 0.001), chest infection (OR = 5.7; 95% CI, 1.8-18.3; p = 0.003) and the hyperglucemia in admission (OR = 6; 95% CI, 1.5-25.6; p = 0.015) are the independient predictor factors that increased the likelihood for mortality in acute stroke. Baseline imbalance NIHSS > 20 (OR = 8.9; 95% CI, 2.7-29; p < 0.001), poor outcome neurological during the first day (OR = 8.1; 95% CI, 2.2 29.6%; p = 0.002) and the urinary incontinence (OR = 10.1; 95% CI, 5-20.6; p < 0.001) are the independient predictor factors that increased the likelihood of poor functional state in discharge. CONCLUSIONS: Dysphagia, crural motor affectation, urinary incontinence and diabetes mellitus are the independient predictor factors that increase the likelihood for nosocomial infection in acute stroke. The chest infection increases significantly the likelihood of mortality during the hospital stay. PMID- 17408537 TI - [Cancer mortality trends in Andalusia (Spain) between 1975 and 2003]. AB - BACKGROUND AND OBJECTIVE: To describe cancer mortality trends in Andalusia for cancer as a whole and for their main locations, by age and gender groups during 1975-2003. MATERIAL AND METHOD: Mortality rates age and gender-adjusted have been estimated through cancer mortality data coming from mortality and polpulation official statistics. Joinpoint regression analysis has been used in order to quantify trends and their points of change. RESULTS: Cancer as a whole has showed a descending trend among women and an ascending trend among men, for the period 1975-2003. Annual change rates were -0.45% and 0.42% for women and men respectively, and both of them, statistically significant (p < 0.05). By age, except for the elder group, a fall is noted, both among women and among men. Among women, breast cancer was the first cause of cancer death; in 1993 it appeared a statistically significant change from ascendent to descendent trend. Among men, lung cancer was the first mortality cause; in 1996 appeared a significant trend change and it began to go down. Colon and rectum cancer was the second one in magnitude, both among men and women, but far from the first one and with an ascendent trend, specially among men. CONCLUSIONS: Cancer mortality shows a descending trend among andalusian women, with a similar rate to the Spanish average. Among men, the trend is ascendent with a higher magnitude than the national average. PMID- 17408538 TI - [Measurement of NT-proBNP in elderly patients with acute dyspnea: diagnostic and pronostic value]. AB - BACKGROUND AND OBJECTIVE: The aim of our study was to establish weather N terminal fragment of brain-type natriuretic peptide (NT-proBNP) is useful in the differential diagnosis of dyspnea in elderly patients, and whether its determination has a prognostic value in heart failure (HF). PATIENTS AND METHOD: 79 patients admitted with acute dyspnea at the emergency department were included in our study. The mean age (standard deviation) was 77.56 (8.71) years. 67% had a diagnosis of HF based on clinical and echocardiagraphic criteria. A follow-up of 18 months was performed after discharge. RESULTS: Higher levels of NT-proBNP were found in patients with HF (6,833.54 pg/ml) than in patients with other causes of dyspnea (1,801.99 pg/ml) (p < 0.0001). A cut-off point of NT-proBNP > 730 pg/ml was related to higher rates of readmission due to HF over the next 18 months. CONCLUSIONS: NT-proBNP is a useful biomarker in the differential diagnosis of dyspnea in the elderly population. Its determination has a prognostic roll, stratifying the risk of readmission in HF patients. PMID- 17408539 TI - [Role of biologic therapy in systemic autoimmune diseases]. PMID- 17408540 TI - [Rituximab for treatment of patients with systemic autoimmune diseases]. AB - BACKGROUND AND OBJECTIVE: To assess the value of rituximab in systemic autoimmune diseases which are refractory to others treatments. PATIENTS AND METHOD: Prospective study on 12 patients -7 with systemic lupus erythematosus (SLE), 4 with Wegener's granulomatosis (WG), and 1 with overlapping connective disease and autoimmune thrombocytopenia-, controlled in a specialized unit of a tertiary hospital. Four weekly doses of rituximab, 2 biweekly doses of cyclophosphamide, and glucocorticoids were administered to all patients, and other immunosuppressants were also administered as considered necessary in each case. RESULTS: Mean follow up after treatment with rituximab was 12.8 moths for SLE patients and 12.3 for WG patients. In SLE patients, proteinuria was reduced below 1 g daily in 5 cases (83%), with a clear parallel improvement in the urinary sediment. Serositis was resolved in both cases. One patient required 3 treatment cycles to obtain an adequate response and another required a second cycle for relapse. Only one patient with WG had a favorable response. The patient treated for autoimmune thrombocytopenia had a favorable response, with no relapses, and creatine-kinase levels also tended to return to normal. There were 2 serious adverse events (terminal renal failure and serious colitis in a patient with SLE, and death of one patient with WG), that were not adjudicated directly to rituximab. Immunoglobulin levels did not change substantially. There were no infusion reactions or associated infections. CONCLUSIONS: Rituximab was useful in patients with SLE refractory to other immunosuppressants. On the contrary, its efficacy in WG was limited. The response of thrombocytopenia was complete and maintained. PMID- 17408541 TI - [Health, economy and freedom: forty years of economic growth, political transition and health conditions in Spain]. PMID- 17408542 TI - [Hereditary breast and ovarian cancer: primary and secondary prevention for BRCA1 and BRCA2 mutation carriers]. AB - Ten years after the identification of the breast and ovarian cancer predisposition genes, BRCA1 and BRCA2, an emerging literature has explored the efficacy of strategies for primary as well as secondary prevention of breast and ovarian cancer in BRCA1 and BRCA2 mutations carriers. The integration of magnetic resonance imaging for breast cancer screening in BRCA carriers has achieved a higher sensibility of the screening, and an early detection of breast cancer. However, we don't have yet enough level of evidence that magnetic resonance imaging could reduce mortality in BRCA carriers. Because of the limitations of screening for ovarian cancer, bilateral prophylactic salpingo-oophorectomy has been established as one of the most effective strategies to prevent ovarian cancer in BRCA1 and BRCA2 mutation carriers. This strategy also has a favorable effect on the reduction of risk of breast cancer if is performed before the age of 50. The management of this patients must be individualized and multidisciplinary. PMID- 17408543 TI - [Comparison of the SCORE and Framingham-REGICOR functions for cardiovascular risk assesment]. PMID- 17408544 TI - [Acute promyelocytic leukemia in a patient with colon adenocarcinoma treated with fluorouracil]. PMID- 17408545 TI - [Hospital medicine, quo vadis?]. PMID- 17408546 TI - [Meningitis as the only manifestation of acute Q fever]. PMID- 17408547 TI - [Chronic tophaceous gout]. PMID- 17408548 TI - [High-grade dysplasia as a risk factor of metachronous advanced colorectal neoplasms in patients with advanced adenomas]. AB - BACKGROUND: Patients with advanced adenomas (AA) have a high risk of developing advanced colorectal neoplasms. Therefore, shorter monitoring intervals have been recommended in this patient subgroup. High grade dysplasia (HGD) is the main marker of cancer transformation. However, its predictive value for developing advanced neoplams in patients with advanced adenoma is unknown. AIM: To investigate if HGD increases the risk for developing advanced neoplasms in patients with AA. METHODS: Between January 1996 and December 1997 every patient with an AA endoscopically resected were considered for inclusion. Patients with a history of colorectal cancer (CRC), inflammatory bowel disease, familial adenomatous polyposis or patients who met the Amsterdam criteria, and those without colonoscopic monitoring were excluded. We assessed the development of advanced neoplasms during the study period. RESULTS: 71 patients were included and classified into 2 groups, depending on the presence (n = 49) or lack (n = 22) of HGD in the initial colonoscopy. The probability of developing advanced neoplasms (log rank, p = 0.47; Breslow, p = 0.58) or AA with HGD (log rank, p = 0.47; Breslow, p = 0.53) in the study period was similar between both groups. The number of metachronic polyps (p = 0.67), adenomas (p = 0.73), AA (p = 0.93) and AA with HGD (p = 0.88) was also similar. CONCLUSION: The risk of developing advanced neoplasms is not different between AA with HGD and those with other characteristics of AA (villous pattern and larger than 1 cm). Therefore, changes in monitoring intervals are not warranted. PMID- 17408549 TI - Psychometric properties of the original Inflammatory Bowel Disease Questionnaire, a Spanish version. AB - BACKGROUND: The Inflammatory Bowel Disease Questionnaire (IBDQ) was developed by Guyatt et al. (1989) and it is the most widely used health-related quality of life instrument for patients with Crohn's disease (CD) and ulcerative colitis (UC). The aim of this study was to assess the psychometric properties of a Spanish version of the original IBDQ by examining the instrument's underlying factor structure, the internal and external validity, and the internal consistency reliability. MATERIAL AND METHOD: One hundred and forty seven patients (76 CD, 71 UC) completed the Spanish version of the IBDQ, the Hospital Anxiety and Depression Scale (HADS) and the Temperament and Character Inventory (TCI). Clinical activity was assessed by the CDAI and the Truelove-Witts index. RESULTS: The confirmatory factor analysis of the IBDQ failed to reproduce the original four-factor structure proposed by Guyatt et al. We found that the Spanish version of the IBDQ consists also of four underlying factors, but the content of each factor and the items included were slightly different. Psychometric testing of the IBDQ revealed that the questionnaire has an acceptable internal and external validity, and a high internal reliability. CONCLUSION: Although confirmatory factor analyses failed to reproduce the original psychometric structure of IBDQ, it seems that the Spanish version of this instrument proved to be valid and reliable for assessing health related quality of life in inflammatory bowel disease patients. PMID- 17408550 TI - Collagenous duodeno-ileo-colitis with transient IgG deficiency preceded by Yersinia enterocolitica intestinal infection: case report and review of literature. AB - A case-report of a man with chronic diarrhoea is presented. After an unsuccessful treatment of an intestinal yersioniosis, the diagnosis of collagenous intestinal disease affecting duodenum, ileum and colon was made. In addition, a IgG transient deficiency was observed. The literature about gastrointestinal involvement, concomintant infection by Yersinia and IgG deficiency in collagenous colitis is reviewed. PMID- 17408551 TI - [Hepatic angiomyolipoma in two patients infected with hepatitis C virus]. AB - AIM: The aim of this study is to present our experience with two cases of hepatic angiomyolioma in hepatitis C virus (HCV) positive patients, and to up-date the clinical manage, diagnostic and treatment of this entity. CLINICAL OBSERVATIONS: Both cases were presented in women in their 4-5th decade of life. Clinical presentation was with symptoms in one but incidental in the other. Both were HCV positive. Values of alpha-fetoprotein were normal. Radiological imaging was not diagnostic. Histopathological examination and immunohistochemical findings gave the diagnosis of angiomyolipoma. At time of diagnosis the size of tumours was 4.8 and 8 cm of diameter. Both cases were treated with surgery in order to definetly rule out malignancy. After 6 and 3 years of follow-up, there is no evidence of recurrence. DISCUSSION AND CONCLUSION: The hepatic angiomyolipoma is a rare benign tumour, mimicking other liver tumours. Although no patognomonic features, there are some radiological findings that point out to the diagnosis of angiomy olipoma. Nevertheless, definitive diagnosis is done by his tological and immunohistochemical findings (HMB-45). The hepatic angiomyolipoma consists of varing proportion of three elements, mature fat cells, smooth muscle cells and blood vessels. Although it is a benign tumour, the difficulty in ruling out malignancy, prompted surgical management. It is not described its relation with HCV virus, thus we consider our cases as an coincident finding. PMID- 17408552 TI - [Obstructive jaundice caused by duodenal amyloidosis succesfully treated with surgery]. AB - We present a case of obstructive jaundice due to duodenal amyloidosis presenting as a bulbar pseudotumor (amyloidoma). The duodenal and biliary obstruction were treated by double bypass, hepatojejunostomy and gastroenterostomy. Our case suggests that surgical palliative treatment may be effective, and that anastomosis is probably safe, allowing the patient a good quality of life. PMID- 17408553 TI - [Filgrastim in refractory Crohn's disease with an intra-abdominal abscess]. AB - The application of recombinant human granulocyte colony-stimulating factor (filgrastim) seems to be a safe, well tolerated and potentially effective therapy for active Crohn's disease. We report the case of an adolescent boy with Crohn's disease and intra-abdominal abscess associated who had a significant response to treatment with recombinant human granulocyte colony-stimulating factor after all standard treatments had failed. PMID- 17408554 TI - [Eosinophilic esophagitis. Review of current clinical and physiopathological concepts]. AB - Eosinophilc esophagitis (EE) is an emerging disease which is characterized by a dense infiltration of the esophagus by eosinophilic leukocytes. The main symptoms of this disease are dysphagia and frequent food impaction in esophagus, and they are due to a hypersentivity response to different foods or aeroallergens. Eosinophil accumulation in the esophageal epithelium is determined by local production of eosinophilotropic cytokines and chemokines, which have been well defined as a TH2-type hypersensitivity reaction in animal models of the disease. Esophageal epithelium, after T CD4+ lymphocytes stimulation, contains all the necessary cell types for the development of local immunoallergic responses. However, there is increasing data on the significant role that humoral immunity could play in the pathophysiology of EE, by means of the action of IgE over mast cells function. The high density of T CD8+ lymphocytes in inflammatory infiltrate suggests that a TH1-type reaction could also participate in the mechanism of the disease. Proteins contained in cytoplasmic granules of activated eosinophils and mast cells could act over neural and muscular components of the esophageal wall, triggering motor disturbances which can be measured by means of manometric recordings and justify the esophageal symptoms. This paper aims to review the newest clinical aspects of EE and the results of studies directed at investigating the pathophysiology of the disease. Furthermore, we carry out a critical review of available therapeutic options. PMID- 17408555 TI - [Pain originating from the abdominal wall: a forgotten diagnostic option]. AB - Chronic abdominal pain is a common clinical problem in primary care, and is usually referred to gastroenterologists or general surgeons. Although up to 20% of cases of idiopathic abdominal pain arise in structures of the abdominal wall, this is frequently overlooked as a possible cause. It includes pain arising from structures of the abdominal wall including skin, parietal peritoneum, cellular subcutaneous tissue, aponeuroses, abdominal muscles and somatosensorial innervation from lower dorsal roots. The diagnosis is based on anamnesis and physical examination. Carnett's sign is a simple maneuver that discriminates between parietal and visceral pain. Management with topical anesthesia is effective in a majority of patients and can help to confirm the diagnosis. PMID- 17408556 TI - [Multiple hepatic abscesses due to Gemella morbillorum]. PMID- 17408557 TI - [Twisted epiploic appendix. A neoplastic recurrence-metastasis simulator]. PMID- 17408558 TI - [Exceptional pancytopenia secondary to treatment with pegylated interferon and ribavirin]. PMID- 17408559 TI - [Ideas, research and ethics of scientific journals]. PMID- 17408560 TI - Santiago Ramon y Cajal at Clark University, 1899; his only visit to the United States. AB - In 1899, at the invitation of G. Stanley Hall, the great psychologist and President of Clark University, Santiago Ramon y Cajal, and four other European scientists of significant note, were invited to participate in the Decennial Celebration of Clark. Cajal, accompanied by his wife, arrived in Worcester, via New York, to much acclaim and praise in the local press. His three lectures, all delivered in French and illustrated with large color drawings made upon his arrival at Clark, were concerned with previously unpublished observations on the structure of the human cerebral cortex. The full text of these lectures and 31 illustrations (in black and white) were published, in English, in a large Decennial Volume prepared by Clark University. At the culmination of the Clark Celebration, Cajal, and the other invited attendees, received the honorary Doctor of Laws degree. Cajal, ever the scholar, visited many sites of interest in the Northeastern US prior to his return to Spain including Columbia, Harvard, and the University of New York. This paper details the events surrounding Cajal's visit to Clark University, his only visit to the United States. PMID- 17408561 TI - Stress and ageing interactions: a paradox in the context of shared etiological and physiopathological processes. AB - Gerontology has made considerable progress in the understanding of the mechanisms underlying the ageing process and age-related neurodegenerative disorders. However, ways to improve quality of life in the elderly remain to be elucidated. It is now clear that stress and the ageing process share a number of underlying mechanisms bound in a very close, if not indissociable, relationship. The ageing process is regulated by the factors underlying the ability to adjust to stress, whilst stress has an influence on the life span and the quality of ageing. In addition, the ability to cope with stress in adulthood predicts life expectancy and quality of life at senescence. The ageing process and stress also share several common mechanisms, particularly in relation to the energy factor. Stress consumes energy and ageing may be considered as a cost of the energy expended to deal with the stressors to which the body is exposed throughout its lifetime. This suggests that the ageing process is associated with and/or a consequence of a long-lasting activation of the major stress responsive systems. However, despite common features, the interaction between stress and the ageing process gives rise to some paradoxes. Stress can either diminish or exacerbate the ageing process just as the ageing process can worsen or counter the effects of stress. There has been little attempt to understand how ageing and stress might interact to promote "successful" or pathological ageing. A key factor in this respect is the individual's ability to adapt to stress. Viewed from this angle, the quality of life of aged subjects may be improved through therapy designed to improve the tolerance to stress. PMID- 17408562 TI - Molecular networks of brain and immunity. AB - Exciting complexity of natural phenomena can be based on rather simple biophysical principles. For example, the genetic code is based on a double-helix of DNA formed by planar geometry of weak hydrogen bounds. On the examples of cytokine networks, immune synapse, psychoneuroimmunology and systems biology, this review paper attempts to show how molecular networks both in brain and immunity can be studied using common principles of protein-protein interactions. PMID- 17408563 TI - Neurotransmitter receptor heteromers and their integrative role in 'local modules': the striatal spine module. AB - 'Local module' is a fundamental functional unit of the central nervous system that can be defined as the minimal portion of one or more neurons and/or one or more glial cells that operates as an independent integrative unit. This review focuses on the importance of neurotransmitter receptor heteromers for the operation of local modules. To illustrate this, we use the striatal spine module (SSM), comprised of the dendritic spine of the medium spiny neuron (MSN), its glutamatergic and dopaminergic terminals and astroglial processes. The SSM is found in the striatum, and although aspects such as neurotransmitters and receptors will be specific to the SSM, some general principles should apply to any local module in the brain. The analysis of some of the receptor heteromers in the SSM shows that receptor heteromerization is associated with particular elaborated functions in this local module. Adenosine A(2A) receptor-dopamine D(2) receptor-glutamate metabotropic mGlu(5) receptor heteromers are located adjacent to the glutamatergic synapse of the dendritic spine of the enkephalin MSN, and their cross-talk within the receptor heteromers helps to modulate postsynaptic plastic changes at the glutamatergic synapse. A(1) receptor-A(2A) receptor heteromers are found in the glutamatergic terminals and the molecular cross-talk between the two receptors in the heteromer helps to modulate glutamate release. Finally, dopamine D(2) receptor-non-alpha(7) nicotinic acetylcholine receptor heteromers, which are located in dopaminergic terminals, introduce the new concept of autoreceptor heteromer. PMID- 17408564 TI - Apoptotic neuronal death in Parkinson's disease: involvement of nitric oxide. AB - Apoptosis of nigral dopaminergic neurons by various mechanisms is an emerging phenomenon involved in the degeneration of dopaminergic neurons in Parkinson's disease (PD). Both extrinsic and intrinsic pathways seems to be involved in death of nigral neurons, intrinsic pathway however, seems to be more important due to the energy crisis. Apoptosis by intrinsic pathway is executed by several initiators and effector caspases, which have been found activated in PD patients, experimental models as well as in neuronal cultures. Nitric oxide (NO) seems to be a central molecule due to its ability to modulate both pro and antiapoptotic phenomenon. The review focuses on the diverse extrinsic and intrinsic factors, signaling pathways and their modulation by NO leading to the death of dopaminergic neurons. PMID- 17408565 TI - The sensory neuron and the triumph of Camillo Golgi. AB - While Golgi's concept of the sensory neuron provided sound reasons for his rejection of the polarity principles underlying the 'neuron doctrine', it is now apparent that his concern about recovery of function after injury and the vast modern findings of ephemerality of connexin-clustered connections in the cerebral cortex and elsewhere in the central nervous system, and credibly termed 'reticularist', has somewhat eclipsed the polarized neuron doctrine of reflex physiology with the "fixed and immutable" connections championed by Cajal. Although Golgi's view was not the result of incisive reasoning based on subsequently confirmed observation, both principles espoused by these combatant Nobel laureate partners have proven robustly operative in different spheres and time frames of neural activity that have vastly enhanced contemporary understanding of neural connectivity. PMID- 17408566 TI - On the role of receptor-receptor interactions and volume transmission in learning and memory. AB - Learning and memory seem to be inherent to a biological neural network. To emerge, they need an extensive functional connectivity, enabling a large repertoire of possible responses to stimuli, and sensitivity of the connectivity to activity, allowing for the selection of adaptive responses. According to the classical view about the organization of the CNS, the connectivity issue is realized by the huge amount of synaptic contacts each neuron establishes, while the adaptation of the network to specific tasks is obtained by mechanisms of activity-dependent synaptic plasticity. The discovery of direct receptor-receptor interactions at the level of the plasma membrane and the existence in the brain of two main modes of communication, the wiring transmission (such as the synaptic transmission) and the volume transmission (based on the diffusion of signals in the extracellular space), provided a broader view of the functional organization of the CNS with potential important consequences on the understanding of learning and memory processes. Owing to receptor-receptor interactions clusters of receptors, the receptor mosaics (RM), can be formed at the plasma membrane where they can work as collective functional units. As a consequence, the connections between the cells become themselves networks (molecular networks) able to adapt their function according to the stimuli they receive. Learning, therefore, can occur also at the level of RMs. Thus, memory formation seems not only to be a distributed process, but also to follow a hierarchical morpho-functional organization. Furthermore, the combination of the two different forms of transmission could allow processes of correlation and coordination to be established between networks and network elements without the need of additional physical connections, leading to a significant increase of the degrees of freedom available to the CNS for learning. PMID- 17408567 TI - Cajal: lessons on brain development. AB - In 1906, Santiago Ramon y Cajal was awarded the Nobel Prize in Physiology or Medicine in recognition of his work on the structure of the nervous system. At that time, almost all of Cajal's work was carried out using the Golgi method, a technique devised by the Italian scientist Camillo Golgi, with whom he shared this prize. Cajal introduced several modifications to the method developed by Golgi and, to avoid the problems encountered in staining myelinated neurons, part of his studies were carried out on embryos and very young animals (the "ontogenetic method"). In this way, Cajal begin to describe aspects of the development of the nervous system. Here, we review some of his wonderful discoveries (for example, the description of the axonal growth cone) from which he derived some of his main theories on the anatomy and physiology of the nervous system: the chemotactic hypothesis and the neuron doctrine. PMID- 17408568 TI - Fly MARCM and mouse MADM: genetic methods of labeling and manipulating single neurons. AB - The Golgi staining method has served neuroscience well for more than a century. In this assay I review recent progresses using genetic methods to recapitulate and extend the Golgi staining method. These methods enable new discoveries on organization and development of neuronal circuits in the fly and mouse brains. PMID- 17408569 TI - Two wrongs make a right: deficits in reversal learning after orbitofrontal damage are improved by amygdala ablation. PMID- 17408570 TI - Astrocytes take center stage in salt sensing. PMID- 17408571 TI - The broader, the better? Drosophila olfactory interneurons are found to respond to a wider range of odorants than their immediate sensory input. PMID- 17408572 TI - Pauses as neural code in the cerebellum. PMID- 17408573 TI - Melting the iceberg: contrast invariance in visual cortex. PMID- 17408574 TI - The scent of Drosophila sex. PMID- 17408575 TI - Molecular neuroanatomy's "Three Gs": a primer. PMID- 17408576 TI - Reversible silencing of neuronal excitability in behaving mice by a genetically targeted, ivermectin-gated Cl- channel. AB - Several genetic strategies for inhibiting neuronal function in mice have been described, but no system that directly suppresses membrane excitability and is triggered by a systemically administered drug, has been validated in awake behaving animals. We expressed unilaterally in mouse striatum a modified heteromeric ivermectin (IVM)-gated chloride channel from C. elegans (GluClalphabeta), systemically administered IVM, and then assessed amphetamine induced rotational behavior. Rotation was observed as early as 4 hr after a single intraperitoneal IVM injection (10 mg/kg), reached maximal levels by 12 hr, and was almost fully reversed by 4 days. Multiple cycles of silencing and recovery could be performed in a single animal. In striatal slice preparations from GluClalphabeta-expressing animals, IVM rapidly suppressed spiking. The two subunit GluCl/IVM system permits "intersectional" strategies designed to increase the cellular specificity of silencing in transgenic animals. PMID- 17408577 TI - Basolateral amygdala lesions abolish orbitofrontal-dependent reversal impairments. AB - Damage to orbitofrontal cortex (OFC) has long been associated with deficits in reversal learning. OFC damage also causes inflexible associative encoding in basolateral amygdala (ABL) during reversal learning. Here we provide a critical test of the hypothesis that the reversal deficit in OFC-lesioned rats is caused by this inflexible encoding in ABL. Rats with bilateral neurotoxic lesions of OFC, ABL, or both areas were tested on a series of two-odor go/no-go discrimination problems, followed by two serial reversals of the final problem. As expected, all groups acquired the initial problems at the same rate, and rats with OFC lesions were slower to acquire the reversals than sham controls. This impairment was abolished by accompanying ABL lesions, while ABL lesions alone had no effect on reversal learning. These results are consistent with the hypothesis that OFC facilitates cognitive flexibility by promoting updating of associative encoding in downstream brain areas. PMID- 17408578 TI - Glial Nax channels control lactate signaling to neurons for brain [Na+] sensing. AB - Sodium (Na) homeostasis is crucial for life, and Na levels in body fluids are constantly monitored in the brain. The subfornical organ (SFO) is the center of the sensing responsible for the control of salt-intake behavior, where Na(x) channels are expressed in specific glial cells as the Na-level sensor. Here, we show direct interaction between Na(x) channels and alpha subunits of Na(+)/K(+) ATPase, which brings about Na-dependent activation of the metabolic state of the glial cells. The metabolic enhancement leading to extensive lactate production was observed in the SFO of wild-type mice, but not of the Na(x)-knockout mice. Furthermore, lactate, as well as Na, stimulated the activity of GABAergic neurons in the SFO. These results suggest that the information on a physiological increase of the Na level in body fluids sensed by Na(x) in glial cells is transmitted to neurons by lactate as a mediator to regulate neural activities of the SFO. PMID- 17408579 TI - Distributed network actions by nicotine increase the threshold for spike-timing dependent plasticity in prefrontal cortex. AB - Nicotine enhances attention and working memory by activating nicotinic acetylcholine receptors (nAChRs). The prefrontal cortex (PFC) is critical for these cognitive functions and is also rich in nAChR expression. Specific cellular and synaptic mechanisms underlying nicotine's effects on cognition remain elusive. Here we show that nicotine exposure increases the threshold for synaptic spike-timing-dependent potentiation (STDP) in layer V pyramidal neurons of the mouse PFC. During coincident presynaptic and postsynaptic activity, nicotine reduces dendritic calcium signals associated with action potential propagation by enhancing GABAergic transmission. This results from a series of presynaptic actions involving different PFC interneurons and multiple nAChR subtypes. Pharmacological block of nAChRs or GABA(A) receptors prevented nicotine's actions and restored STDP, as did increasing dendritic calcium signals with stronger postsynaptic activity. Thus, by activating nAChRs distributed throughout the PFC neuronal network, nicotine affects PFC information processing and storage by increasing the amount of postsynaptic activity necessary to induce STDP. PMID- 17408580 TI - Excitatory interactions between olfactory processing channels in the Drosophila antennal lobe. AB - Each odorant receptor gene defines a unique type of olfactory receptor neuron (ORN) and a corresponding type of second-order neuron. Because each odor can activate multiple ORN types, information must ultimately be integrated across these processing channels to form a unified percept. Here, we show that, in Drosophila, integration begins at the level of second-order projection neurons (PNs). We genetically silence all the ORNs that normally express a particular odorant receptor and find that PNs postsynaptic to the silent glomerulus receive substantial lateral excitatory input from other glomeruli. Genetically confining odor-evoked ORN input to just one glomerulus reveals that most PNs postsynaptic to other glomeruli receive indirect excitatory input from the single ORN type that is active. Lateral connections between identified glomeruli vary in strength, and this pattern of connections is stereotyped across flies. Thus, a dense network of lateral connections distributes odor-evoked excitation between channels in the first brain region of the olfactory processing stream. PMID- 17408581 TI - A parturition-associated nonsynaptic coherent activity pattern in the developing hippocampus. AB - Correlated neuronal activity is instrumental in the formation of networks, but its emergence during maturation is poorly understood. We have used multibeam two photon calcium microscopy combined with targeted electrophysiological recordings in order to determine the development of population coherence from embryonic to postnatal stages in the hippocampus. At embryonic stages (E16-E19), synchronized activity is absent, and neurons are intrinsically active and generate L-type channel-mediated calcium spikes. At birth, small cell assemblies coupled by gap junctions spontaneously generate synchronous nonsynaptic calcium plateaus associated to recurrent burst discharges. The emergence of coherent calcium plateaus at birth is controlled by oxytocin, a maternal hormone initiating labour, and progressively shut down a few days later by the synapse-driven giant depolarizing potentials (GDPs) that synchronize the entire network. Therefore, in the developing hippocampus, delivery is an important signal that triggers the first coherent activity pattern, which is silenced by the emergence of synaptic transmission. PMID- 17408582 TI - Cerebellar LTD and pattern recognition by Purkinje cells. AB - Many theories of cerebellar function assume that long-term depression (LTD) of parallel fiber (PF) synapses enables Purkinje cells to learn to recognize PF activity patterns. We have studied the LTD-based recognition of PF patterns in a biophysically realistic Purkinje-cell model. With simple-spike firing as observed in vivo, the presentation of a pattern resulted in a burst of spikes followed by a pause. Surprisingly, the best criterion to distinguish learned patterns was the duration of this pause. Moreover, our simulations predicted that learned patterns elicited shorter pauses, thus increasing Purkinje-cell output. We tested this prediction in Purkinje-cell recordings both in vitro and in vivo. In vitro, we found a shortening of pauses when decreasing the number of active PFs or after inducing LTD. In vivo, we observed longer pauses in LTD-deficient mice. Our results suggest a novel form of neural coding in the cerebellar cortex. PMID- 17408583 TI - The emergence of contrast-invariant orientation tuning in simple cells of cat visual cortex. AB - Simple cells in primary visual cortex exhibit contrast-invariant orientation tuning, in seeming contradiction to feed-forward models that rely on lateral geniculate nucleus (LGN) input alone. Contrast invariance has therefore been thought to depend on the presence of intracortical lateral inhibition. In vivo intracellular recordings instead suggest that contrast invariance can be explained by three properties of the excitatory pathway. (1) Depolarizations evoked by orthogonal stimuli are determined by the amount of excitation a cell receives from the LGN, relative to the excitation it receives from other cortical cells. (2) Depolarizations evoked by preferred stimuli saturate at lower contrasts than the spike output of LGN relay cells. (3) Visual stimuli evoke contrast-dependent changes in trial-to-trial variability, which lead to contrast dependent changes in the relationship between membrane potential and spike rate. Thus, high-contrast, orthogonally oriented stimuli that evoke significant depolarizations evoke few spikes. Together these mechanisms, without lateral inhibition, can account for contrast-invariant stimulus selectivity. PMID- 17408584 TI - Encoding of illusory continuity in primary auditory cortex. AB - When interfering objects occlude a scene, the visual system restores the occluded information. Similarly, when a sound of interest (a "foreground" sound) is interrupted (occluded) by loud noise, the auditory system restores the occluded information. This process, called auditory induction, can be exploited to create a continuity illusion. When a segment of a foreground sound is deleted and loud noise fills the missing portion, listeners incorrectly report hearing the foreground continuing through the noise. Here we reveal the neurophysiological underpinnings of illusory continuity in single-neuron responses from awake macaque monkeys' primary auditory cortex (A1). A1 neurons represented the missing segment of occluded tonal foregrounds by responding to discontinuous foregrounds interrupted by intense noise as if they were responding to the complete foregrounds. By comparison, simulated peripheral responses represented only the noise and not the occluded foreground. The results reveal that many A1 single neuron responses closely follow the illusory percept. PMID- 17408585 TI - Learning-related human brain activations reflecting individual finances. AB - A basic tenet of microeconomics suggests that the subjective value of financial gains decreases with increasing assets of individuals ("marginal utility"). Using concepts from learning theory and microeconomics, we assessed the capacity of financial rewards to elicit behavioral and neuronal changes during reward predictive learning in participants with different financial backgrounds. Behavioral learning speed during both acquisition and extinction correlated negatively with the assets of the participants, irrespective of education and age. Correspondingly, response changes in midbrain and striatum measured with functional magnetic resonance imaging were slower during both acquisition and extinction with increasing assets and income of the participants. By contrast, asymptotic magnitudes of behavioral and neuronal responses after learning were unrelated to personal finances. The inverse relationship of behavioral and neuronal learning speed with personal finances is compatible with the general concept of decreasing marginal utility with increasing wealth. PMID- 17408586 TI - The clinical diagnosis of microbial keratitis. AB - PURPOSE: To evaluate the ability of ophthalmologists to predict the laboratory results of presumed microbial keratitis and to explore which findings may influence diagnostic prognostication. DESIGN: Prospective, cross-sectional study. METHODS: Fifteen ophthalmologists completed study forms at the initial presentation of patients with presumed microbial keratitis. After predicting the category of microbial recovery, clinicians submitted corneal scrapings for masked laboratory processing. The relative effects of ocular inflammatory signs on correct microbial diagnosis were explored with Poisson regression. RESULTS: Clinical examiners correctly predicted the presence or absence of microbial recovery in 79 (76%) of 104 ulcerative keratitis cases and successfully distinguished among bacterial, fungal, and amebic keratitis for 54 (73%) of 74 culture-positive infections, although only 31 (42%) were subcategorized properly. The positive predictive value of clinical diagnosis was 65% (95% confidence interval [CI], 43% to 84%) for 20 eyes with Pseudomonas keratitis cases, 48% (95% CI, 32% to 63%) for 38 other bacterial keratitis, 45% (95% CI, 17% to 77%) for 13 fungal keratitis, and 89% (95% CI, 52% to 100%) for nine Acanthamoeba keratitis cases. The recognition of Pseudomonas keratitis significantly improved by the occurrence of a larger infiltrate (P = .02), and correctly predicting Acanthamoeba keratitis was enhanced by observing a ring infiltrate (P < .001). Antimicrobial use before referral significantly attenuated clinical diagnosis (P = .03) and hampered microbial recovery (P = .004). CONCLUSIONS: Established Pseudomonas keratitis and Acanthamoeba keratitis can be suspected before laboratory confirmation, but overlapping inflammatory features and recent empiric antimicrobial treatment limits etiologic recognition of most microbial corneal infections. PMID- 17408587 TI - Electron and nuclear dynamics in many-electron atoms, molecules and chlorophyll protein complexes: a review. AB - It has been shown [V.A. Shuvalov, Quantum dynamics of electrons in many-electron atoms of biologically important compounds, Biochemistry (Mosc.) 68 (2003) 1333 1354; V.A. Shuvalov, Quantum dynamics of electrons in atoms of biologically important molecules, Uspekhi biologicheskoi khimii, (Pushchino) 44 (2004) 79-108] that the orbit angular momentum L of each electron in many-electron atoms is L=mVr=nPlanck's and similar to L for one-electron atom suggested by N. Bohr. It has been found that for an atom with N electrons the total electron energy equation E=-(Z(eff))(2)e(4)m/(2n(2)Planck's(2)N) is more appropriate for energy calculation than standard quantum mechanical expressions. It means that the value of L of each electron is independent of the presence of other electrons in an atom and correlates well to the properties of virtual photons emitted by the nucleus and creating a trap for electrons. The energies for elements of the 1st up to the 5th rows and their ions (total amount 240) of Mendeleev' Periodical table were calculated consistent with the experimental data (deviations in average were 5 x 10(-3)). The obtained equations can be used for electron dynamics calculations in molecules. For H(2) and H(2)(+) the interference of electron-photon orbits between the atoms determines the distances between the nuclei which are in agreement with the experimental values. The formation of resonance electron-photon orbit in molecules with the conjugated bonds, including chlorophyll-like molecules, appears to form a resonance trap for an electron with E values close to experimental data. Two mechanisms were suggested for non barrier primary charge separation in reaction centers (RCs) of photosynthetic bacteria and green plants by using the idea of electron-photon orbit interference between the two molecules. Both mechanisms are connected to formation of the exciplexes of chlorophyll-like molecules. The first one includes some nuclear motion before exciplex formation, the second one is related to the optical transition to a charge transfer state. PMID- 17408588 TI - Salt stress impact on the molecular structure and function of the photosynthetic apparatus--the protective role of polyamines. AB - In the present study the green alga Scenedesmus obliquus was used to assess the effects of high salinity (high NaCl-concentration) on the structure and function of the photosynthetic apparatus and the possibility for alleviation by exogenous putrescine (Put). Chlorophyll fluorescence data revealed the range of the changes induced in the photosynthetic apparatus by different NaCl concentrations, which altogether pointed towards an increased excitation pressure. At the same time, changes in the levels of endogenous polyamine concentrations, both in cell and in isolated thylakoid preparations were also evidenced. Certain polyamine changes (Put reduction) were correlated with changes in the structure and function of the photosynthetic apparatus, such as the increase in the functional size of the antenna and the reduction in the density of active photosystem II reaction centers. Thus, exogenously added Put was used to compensate for this stress condition and to adjust the above mentioned changes, so that to confer some kind of tolerance to the photosynthetic apparatus against enhanced NaCl-salinity and permit cell growth even in NaCl concentrations that under natural conditions would be toxic. PMID- 17408589 TI - Characterization of the conformational and orientational dynamics of ganglioside GM1 in a dipalmitoylphosphatidylcholine bilayer by molecular dynamics simulations. AB - The structure and dynamics of a single GM1 (Gal5-beta1,3-GalNAc4-beta1,4-(NeuAc3 alpha2,3)-Gal2-beta1,4-Glc1-beta1,1-Cer) embedded in a DPPC bilayer have been studied by MD simulations. Eleven simulations, each of 10 ns productive run, were performed with different initial conformations of GM1. Simulations of GM1-Os in water and of a DPPC bilayer were also performed to delineate the effects of the bilayer and GM1 on the conformational and orientational dynamics of each other. The conformation of the GM1 headgroup observed in the simulations is in agreement with those reported in literature; but the headgroup is restricted when embedded in the bilayer. NeuAc3 is the outermost saccharide towards the water phase. Glc1 and Gal2 prefer a parallel, and NeuAc3, GalNac4 and Gal5 prefer a perpendicular, orientation with respect to the bilayer normal. The overall characteristics of the bilayer are not affected by the presence of GM1; however, GM1 does influence the DPPC molecules in its immediate vicinity. The implications of these observations on the specific recognition and binding of GM1 embedded in a lipid bilayer by exogenous proteins as well as proteins embedded in lipids have been discussed. PMID- 17408590 TI - An investigation of self-ambivalence in obsessive-compulsive disorder. AB - Individuals vulnerable to obsessive-compulsive disorder (OCD) are hypothesized to have ambivalence about their self-worth, morality and lovability [Guidano, V., & Liotti, G. (1983). Cognitive processes and emotional disorders. New York: Guildford Press]. The purpose of this study was to examine whether self ambivalence was associated with OCD phenomena and beliefs relevant to OCD. It also examined whether patients with OCD had higher levels of self-ambivalence compared to non-clinical controls and patients with other anxiety disorders. Participants were 73 OCD patients, 50 patients with another anxiety disorder, 225 non-clinical undergraduate students and 43 non-clinical community controls. They completed measures of self-ambivalence, OCD phenomena, OCD-relevant beliefs, depression, anxiety and self-esteem. Self-ambivalence was significantly associated with OCD phenomena and OCD-relevant beliefs, after controlling for self-esteem, depression and anxiety. Further, OCD participants were significantly more ambivalent than the non-clinical groups, but did not differ from anxious controls. It was argued that these results provide a basis for extending the cognitive-behavioural model of OCD to include ambivalent self-perceptions as a component of the cognitive mechanisms relevant to the disorder. PMID- 17408591 TI - Regulation of human umbilical cord blood-derived multi-potent stem cells by autogenic osteoclast-based niche-like structure. AB - Stem cell niches provide the micro-environment for the development of stem cells. Under our culturing regimen, a kind of osteoclast-centralized structure supports the proliferation of MSCs, derived from human cord blood, once they reside on osteoclasts. MSCs in this structure expressed Oct4 which is a marker of embryonic stem cells. Floating daughter cells of MSCs colony showed abilities to differentiate into osteocyte, adipocyte, and neuronal progenitor cells. Compared with the easy senescence of MSCs without this niche-like structure in vitro, these results suggested that osteoclasts might play an important role the development and maintenance of Umbilical cord blood (UCB)-derived MSCs and might provide a means to expand UCB-MSCs in vitro, more easily, through a stem cell niche-like structure. PMID- 17408592 TI - GTP/GDP binding stabilizes bacterial cell division protein FtsZ against degradation by FtsH protease in vitro. AB - Factors contributing to the stability of bacterial cell division protein FtsZ remain unknown. In order to identify FtsZ-stabilizing factor(s), we exploited FtsH protease-based in vitro FtsZ degradation assay system. Whole cell lysate from an ftsH-null strain of Escherichia coli inhibited degradation of FtsZ by FtsH in vitro. However, activated charcoal-treated lysate did not inhibit degradation. The loss of ability of the activated charcoal-treated lysate to inhibit degradation of FtsZ was restored when it was replenished with GTP, but not when replenished with other NTPs or dNTPs. The lysate did not protect either FtsZ deletion mutants, which do not bind GTP, or FtsH substrates, sigma(32) and cI-108 proteins, against FtsH. GDP and GTPgammaS also stabilized FtsZ against FtsH. Neither GTP nor GDP inhibited proteolytic activity of FtsH per se. These observations demonstrate that binding of GTP/GDP ligands is responsible for the proteolytic stability of FtsZ against FtsH. PMID- 17408593 TI - Role of myofibril-inducing RNA in cardiac TnT expression in developing Mexican axolotl. AB - The Mexican axolotl, Ambystoma mexicanum, has been a useful animal model to study heart development and cardiac myofibrillogenesis. A naturally-occurring recessive mutant, gene "c", for cardiac non-function in the Mexican axolotl causes a failure of myofibrillogenesis due to a lack of tropomyosin expression in homozygous mutant (c/c) embryonic hearts. Myofibril-inducing RNA (MIR) rescues mutant hearts in vitro by promoting tropomyosin expression and myofibril formation thereafter. We have studied the effect of MIR on the expression of various isoforms of cardiac troponin T (cTnT), a component of the thin filament that binds with tropomyosin. Four alternatively spliced cTnT isoforms have been characterized from developing axolotl heart. The expression of various cTnT isoforms in normal, mutant, and mutant hearts corrected with MIR, is evaluated by real-time RT-PCR using isoform specific primer pairs; MIR affects the total transcription as well as the splicing of the cTnT in axolotl heart. PMID- 17408594 TI - Visfatin promotes angiogenesis by activation of extracellular signal-regulated kinase 1/2. AB - Adipose tissue is highly vascularized and requires the angiogenic properties for its mass growth. Visfatin has been recently characterized as a novel adipokine, which is preferentially produced by adipose tissue. In this study, we report that visfatin potently stimulates in vivo neovascularization in chick chorioallantoic membrane and mouse Matrigel plug. We also demonstrate that visfatin activates migration, invasion, and tube formation in human umbilical vein endothelial cells (HUVECs). Moreover, visfatin evokes activation of the extracellular signal regulated kinase 1/2 (ERK1/2) in endothelial cells, which is closely linked to angiogenesis. Inhibition of ERK activation markedly decreases visfatin-induced tube formation of HUVECs and visfatin-stimulated endothelial cell sprouting from rat aortic rings. Taken together, these results demonstrate that visfatin promotes angiogenesis via activation of mitogen-activated protein kinase ERK dependent pathway and suggest that visfatin may play important roles in various pathophysiological angiogenesis including adipose tissue angiogenesis. PMID- 17408595 TI - A molecular dynamics study of WPD-loop flexibility in PTP1B. AB - Protein tyrosine phosphatase 1B (PTP1B) is an important drug target for the treatment of type II diabetes and obesity. There are strong indications that a novel class of allosteric inhibitors act by preventing the closure of the WPD loop [C. Wiesmann, K.J. Barr, J. Kung, J. Zhu, D.A. Erlanson, W. Shen, B.J. Fahr, M. Zhong, L. Taylor, M. Randall, R.S. McDowell, S.K. Hansen, Allosteric inhibition of protein tyrosine phosphatase 1B, Nat. Struc. Mol. Biol. 11 (2004) 730-737.], which is absolutely essential for the catalytic activity of PTP1B. In this work, we develop force field parameters for one of these inhibitors (BB3), and subsequently utilise standard and targeted molecular dynamics simulations to perform a study of WPD-loop mobility in the presence of this inhibitor. We demonstrate that BB3 not only significantly reduces the flexibility of the WPD loop compared to both the apo-enzyme or the closed conformation complexed with phosphotyrosine, but that this is accompanied by reduced flexibility in a related region, the S-loop, further emphasising the possibility of manipulating this region when designing novel inhibitors for PTP1B. PMID- 17408596 TI - Acute blood-brain barrier permeabilization in rats after systemic Phoneutria nigriventer venom. AB - A highly controlled transport of substances at the interface between blood and brain characterizes the blood-brain barrier (BBB), fundamental for maintenance of the homeostasis of the cerebral milieu. In this study, we investigated the time course (15 min, 1, 2, and 5 h) of BBB opening induced by intravenous (i.v.) injection of Phoneutria nigriventer spider venom (PNV) using quantitative and morphological approaches on cerebellum and hippocampus vessels for assessment of BBB permeability. The results showed vasogenic edema and tracer extravasation faster and severalfold higher in hippocampus than in cerebellum. Reactive astrocytes with swollen perivascular end-feet processes were found only in cerebellum. An immediate and total degradation of laminin in capillaries occurred resulting in the disappearance of the basement membrane. In medium-sized vessels, this effect was less prominent. The changes were transient, with cerebellum in general presenting a faster recovery. However, at 5 h laminin was overexpressed, principally in hippocampus. The rapid and abrupt shift of laminin expression in capillaries (at 15 min) coincided with the immediate and severe signs of intoxication shown by the animals, but not with the peak of leakage of vessels and vasogenic edema, which occurred later (1-2 h). The findings suggest a complex regulatory mechanism, since the extension of BBB impairment caused by PNV depends on the region of the SNC, and on the vessels types. It is suggested that the components of the BBB (gliovascular unit) have a critical role in these differences. P. nigriventer venom can be a useful tool to explore the mechanisms of BBB. PMID- 17408597 TI - Phospholipid mediated plasticity in exocytosis observed in PC12 cells. AB - Membrane composition serves to identify intracellular compartments, signal cell death, as well as to alter a cell's electrical and physical properties. Here we use amperometry to show that supplementation with the phospholipids phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelin (SM), and phosphatidylserine (PS) can alter several aspects of exocytosis. Changes in the amperometric peak shape derived from individual exocytosing vesicles reveal that PC slows expulsion of neurotransmitter while PE accelerates expulsion of neurotransmitter. Amperometry data reveal a reduced amount of catecholamine released per event from PC-treated cells while electron micrographs indicate the vesicles in these cells are 50% larger than controls, thus providing evidence of pharmacological changes in vesicle concentration. Addition of SM appears to affect the rate of fusion pore expansion, indicated by slower peak rise times, but does not affect decay times or quantal size. Addition of PS results in a 1.7 fold increase in the number of events elicited by high-K(+) depolarization. Electron micrographs of PS-treated cells suggest that increased vesicle recruitment underlies enhanced secretion. We did not observe any effect of phosphatidylinositol (PI) treatment. Together these data suggest that differences in membrane composition affect exocytosis and might be involved in mechanisms of cell function controlling the dynamics of communication via exocytosis. PMID- 17408598 TI - Anticipation of affective images and event-related desynchronization (ERD) of alpha activity: an MEG study. AB - We investigated the event-related power decrease (event-related desynchronization: ERD) of the alpha bands associated with the anticipation of affective images. Participants (n=19) were presented with emotionally positive or negative images under different anticipatory conditions, and their brain responses were recorded using magnetoencephalography (MEG). In the Affective Cue conditions, the cue stimulus indicated the emotional valence (positive or negative) of the image. In the Null Cue condition, the cue stimulus did not include any information about the valence of the image, and in the No Cue condition, the affective image was presented without a preceding cue. The cues in the affective and null conditions were followed by emotional images. During the anticipation period for the affective image, the alpha ERD preceding an anticipated negative image was larger than that preceding an anticipated positive image; this effect had an occipital dominance. Furthermore, during the anticipation period, the lower-2-alpha ERD of the right frontal area showed the same result. These results demonstrate that anticipation of negative stimuli induced alpha ERD in both the visual and the right frontal cortex, indicating that top-down modulation may be provided by the right frontal cortex to the visual cortex. PMID- 17408599 TI - Effect of levetiracetam on molecular regulation of hippocampal glutamate and GABA transporters in rats with chronic seizures induced by amygdalar FeCl3 injection. AB - Enhancement of the glutamatergic excitatory synaptic transmission efficacy in the FeCl3 induced epilepsy model is associated with changes in the levels of glutamate and GABA transporter proteins. This study examined the effect of levetiracetam (LEV) on glutamate overflow and glutamate/GABA transporters expression in rats with epileptogenesis induced by the amygdalar injection of 1.0 microl of 100 mM FeCl3 (epileptic rat) and in control rats receiving amygdalar acidic saline injection (non-epileptic rat). In amygdalar acidic saline injected rats, 40 mM KCl-evoked glutamate overflow was significantly suppressed by both 32 and 100 microM LEV co-perfusion. In unilateral amygdalar FeCl3 injected rats, 32 microM LEV was ineffective, but the 100 microM LEV statistically suppressed glutamate overflow. Western blotting was employed to determine the hippocampal expression of glutamate/GABA transporters in epileptic or non-epileptic rats. The rats were treated for 14 days with 54 mg/kg LEV or vehicle intraperitoneally injection. Following 14 days of treatment, the ipsilateral hippocampus was removed for a Western blot analysis. In non-epileptic rats, the expression increased for all of the glutamate and GABA transporters (GLAST, GLT-1, EAAC-1, GAT-1 and GAT-3) while the glutamate transporter regulating protein (GTRAP3-18) decreased in comparison to those of normal rats that were treated with the vehicle. In epileptic rats receiving LEV, the EAAC-1 and GAT-3 levels increased while GTRAP3-18 (89%) decreased in comparison to those of the epileptic rats treated with the vehicle. GTRAP3-18 inhibitor regulates glutamate-binding affinity to EAAC-1. The anti-epileptic action of LEV may be partially due to a reduction of glutamate-induced excitotoxicity and an enhancement of the GABAergic inhibition as observed with the inhibitory effect on the 40 mM KCl-evoked glutamate overflow. These conclusions are supported by the increase in the expression of glial glutamate transporters (GLAST and GLT-1), and the increase in the expression of EAAC-1 and GAT-3 associated with a decrease in GTRAP3-18. The increased expression of EAAC-1 and the decreased expression of GTRAP3-18 in association with the up-regulation of GAT-3 due to such continual LEV administration was thus found to enhance GABA synthesis and reverse the transport of GABA both in non-epileptic and epileptic rats. The suppression of glutamate excitation and the enhancement of GABA inhibition in the rats with continual LEV administration is a result of the up-regulation of glutamate and GABA transporters with the down-regulation of GTRAP3-18. These observations together demonstrated the critical molecular mechanism of the anti-epileptic activity of LEV. PMID- 17408600 TI - The conformation of the C-glycosyl analogue of N-acetyl-lactosamine in the free state and bound to a toxic plant agglutinin and human adhesion/growth-regulatory galectin-1. AB - The conformational behavior of the C-glycoside analogue of N-acetyl-lactosamine, beta-C-Gal-(1-->4)-beta-GlcNAc-OMe, 1, has been studied using a combination of molecular mechanics calculations and NMR spectroscopy (J and NOE data). It is shown that the C-disaccharide populates three distinctive conformational families in solution, the major one being the anti-psi conformation. Of note, this conformation is only marginally populated for the O-disaccharide. Due to its conspicuous role in the regulation of adhesion, growth and tissue invasion of tumors and its avid binding to N-acetyl-lactosamine human, galectin-1 was tested as a receptor. This endogenous lectin recognizes a local minimum of 1, the syn PhiPsi conformer, and thus a conformational selection process is correlated with the molecular recognition event. PMID- 17408601 TI - Extracellular ATP induces assembly and activation of the myosin light chain phosphatase complex in endothelial cells. AB - OBJECTIVES: Extracellular ATP stabilizes the endothelial barrier and inactivates the contractile machinery of endothelial cells. This inactivation relies on dephosphorylation of the regulatory myosin light chain (MLC) due to an activation of the MLC phosphatase (MLCP). To date, activation and function of MLCP in endothelial cells are only partially understood. METHODS: Here, the mechanism of extracellular ATP-mediated activation of MLCP was analyzed in human endothelial cells from umbilical veins. Cells were transfected with the endogenous protein phosphatase 1 (PP1)-specific inhibitor-2 (I-2). RESULTS: Overexpression of I-2 led to inhibition of PP1 activity and abrogation of the ATP-induced dephosphorylation of MLC. This indicates that the PP1 catalytic subunit is the principal phosphatase catalyzing the MLC dephosphorylation induced by extracellular ATP. As demonstrated by immunoprecipitation analysis, extracellular ATP recruits the PP1delta catalytic subunit and the myosin phosphatase targeting subunit (MYPT1) to form a complex. ATP stimulated dephosphorylation of MYPT1 at the inhibitory phosphorylation sites threonine 850 and 696. However, extracellular ATP failed to stimulate MYPT1 dephosphorylation in I-2 overexpressing cells. CONCLUSIONS: The present study shows for the first time that, in endothelial cells, extracellular ATP causes activation of MLCP through recruitment of PP1delta and MYPT1 into a MLCP holoenzyme complex and PP1-mediated reduction of the inhibitory phosphorylation of MYPT1. PMID- 17408602 TI - Nitric oxide stimulates heme oxygenase-1 gene transcription via the Nrf2/ARE complex to promote vascular smooth muscle cell survival. AB - OBJECTIVE: Previous studies from our laboratory and others found that NO is a potent inducer of heme oxygenase-1 (HO-1) gene transcription in vascular smooth muscle cells (SMC), however, the mechanism responsible for the induction of HO-1 gene expression has not been elucidated. In the present study, we determined the signaling pathway responsible for the induction of HO-1 and its biological significance. METHODS: Cultured rat aortic SMC were exposed to nitrosative stress by treating cells with various NO donors or with inflammatory cytokines. RESULTS: Nitrosative stress stimulated an increase in HO-1 mRNA expression and promoter activity in vascular SMC. However, mutation of the antioxidant response element (ARE) in the HO-1 promoter or overexpression of a dominant-negative mutant of NF E2-related factor-2 (Nrf2) abrogated the activation by NO. Electromobility shift assays using an ARE probe detected a complex that was significantly increased in intensity by NO. In addition, the migration of this complex was retarded by using an antibody directed against Nrf2. NO also increased Nrf2 mRNA expression, total and nuclear Nrf2 levels, and the binding of Nrf2 to the HO-1 promoter. Finally, treatment of SMC with NO stimulated apoptosis that was increased by HO-1 inhibition. CONCLUSIONS: These results demonstrate that nitrosative stress induces HO-1 gene transcription through the activation of the Nrf2/ARE complex to counteract NO-induced apoptosis of vascular SMC. The capacity of nitrosative stress to activate Nrf2 and stimulate HO-1 gene transcription may represent a critical adaptive response to maintain cell viability at sites of vascular inflammation and atherosclerosis. PMID- 17408603 TI - CD4 ligation induces activation of protein kinase C zeta and phosphoinositide dependent-protein kinase-1, two kinases required for down-regulation of LFA-1 mediated adhesion. AB - We previously showed that CD4 binding induced a down-regulation of LFA-1 dependent-antigen-independent adhesion of T and B lymphocytes in a phosphatidylinositol-3-kinase (PI3K)-dependent manner. We now show in A201-CD4 (+) T cell lines, that anti-CD4 Ab increases activation of phosphoinositide dependent-protein-kinase 1 (PDK1) or PKC zeta, two main effectors down-stream from PI3K. CD4 binding also increases interactions between PI3K and activated PKCzeta and PDK1. Both events are dependent on CD4/p56Lck association, since they are not detected when p56Lck is unable to bind a truncated form of CD4 in transfected T cell lines. We also show using antisense oligonucleotides that both kinases are necessary for down-regulating LFA-1-dependent adhesion induced by CD4 signalling. We also suggest a role of PDK1 in the recruitment of the phosphatase SHP-2 in a multiprotein complex induced by anti-CD4 Ab. This study thus provides further insights into the mechanism underlying the CD4 triggered regulation of LFA-1-mediated adhesion. PMID- 17408604 TI - Molecular cytotoxic mechanisms of catecholic polychlorinated biphenyl metabolites in isolated rat hepatocytes. AB - Polychlorinated biphenyl (PCB) and PCB metabolites are highly lipophilic and accumulate easily in the lipid bilayer and fat deposits of the body. The molecular cytotoxic mechanisms of these metabolites are still not understood. The aim of the present study was to compare the cytotoxicity and toxicological properties of six dihydroxylated metabolites using isolated rat hepatocytes. All of the metabolites were more cytotoxic than 4-chlorobiphenyl (4-ClBP) and less cytotoxic than phenyl hydroquinone (PHQ). The order of cytotoxic effectiveness of catecholic metabolites expressed as LC(50) (2h) was 3',4'-diCl-2,3-diOH biphenyl>PHQ>4'-Cl-2,5-diOH-biphenyl, 4'-Cl-2,3-diOH-biphenyl>2',5'-diCl-3,4-diOH biphenyl>2',3'-diCl-3,4-diOH-biphenyl>3',4'-diCl-3,4-diOH-biphenyl>4'Cl-3,4-diOH biphenyl>4'-Cl-biphenyl; showing that the positions of hydroxyl and chlorine groups were important for their hepatotoxicity and that the two 2,3-diOH congeners were the most cytotoxic. Cytotoxicity for 3,4-diOH metabolites correlated with the number and position of chlorine atoms with the more chlorine atoms being more cytotoxic. The cytotoxic order of metabolites with two chlorine atoms being 2',5'>2',3'>3',4'. Borneol, an uridine diphosphate glucuronosyltransferases (UGT) inhibitor, increased the cytotoxicity of all tested metabolites; suggesting that glucuronidation was a major mechanism of elimination of these compounds. On the other hand entacapone, a catechol-O-methyl transferase (COMT) inhibitor, only increased the cytotoxicity of 3',4'-diCl-3,4 diOH-biphenyl, 3',4'-diCl-2,3-diOH-biphenyl and 4'-Cl-2,3-diOH-biphenyl. Hepatocyte GSH was depleted (oxidized and conjugated) by these metabolites before cytotoxicity ensued in a similar order of effectiveness to their cytotoxicity with PHQ being the most effective. Hepatocyte mitochondrial membrane potential also decreased before cytotoxicity ensued with a similar order of effectiveness as their cytotoxicity. These results suggest that catecholic cytotoxicity can be attributed to mitochondrial toxicity and oxidative stress. Semiquinone or benzoquinone species were also important in the cytotoxicity of catecholic metabolites. PMID- 17408605 TI - Monitoring of Gaucher patients with a novel chitotriosidase assay. AB - BACKGROUND: Chitotriosidase (CT) is a surrogate plasma marker for Gaucher disease. The enzyme is released by storage cells and is on average thousand fold elevated in serum of Gaucher patients. Plasma CT level is measured with the substrate 4-methylumbelliferyl (4MU)-chitotriose or 4MU-chitobiose. Given the limitations associated with the use of these substrates, a novel substrate, 4MU deoxychitobiose, has recently been conceived. METHODS: Chitotriosidase activity was measured with all three substrates in serum samples from 91 type 1 Gaucher patients. Glucocerebrosidase and chitotriosidase genotypes were determined as well as disease parameters. RESULTS: Chitotriosidase activity when measured with 4MU-deoxychitobiose gave higher values and was proportional to enzyme concentration over a much larger range as compared to the other two substrates. Patients that were carrier for the common CT mutation showed on average half the activity of those with wild type CT genotype. Plasma CT levels correlated best with combined liver and spleen volume: r=0.53 (p<0.001). CONCLUSIONS: The use of 4MU-deoxychitobiose as substrate renders a substantial improved CT activity assay and may further facilitate accurate laboratory monitoring of Gaucher patients. PMID- 17408606 TI - Prognostic significance of interleukin-6 measurement in the diagnosis of acute myocardial infarction in emergency department. AB - BACKGROUND: Markers of inflammation may predict both coronary artery disease (CAD) and adverse outcomes in patients with known CAD. Here, we investigated the role of interleukin-6 (IL-6) in the "triage" and risk assessment of patients admitted to emergency department (ED). METHODS: Serum IL-6 and high sensitivity C reactive protein (hs-CRP) levels were prospectively evaluated in 88 patients with a history of precordial chest pain or shortness of breath of recent onset (<6 h). RESULTS: Of the 88 patients, 21% were discharged from the ED with diagnosis of non-ischemic chest pain (NICP), 39% had a final diagnosis of unstable angina (UA) and 40% experienced an acute myocardial infarction (AMI). Median IL-6 (p<0.001) and hs-CRP (p<0.01) levels on admission were significantly increased in patients with AMI compared with patients with NICP or UA. IL-6 levels correlated with hs CRP (p<0.01). Multivariate analyses including known risk factors showed that elevated creatine kinase-MB (p<0.05) and IL-6 levels (p<0.01) were independently associated with a final diagnosis of AMI. Elevated IL-6 levels significantly predicted the risk of AMI (OR=2.47, p=0.006) in chest pain-enzyme negative patients. CONCLUSIONS: IL-6 may behave as an adjunctive diagnostic tool to assist in the risk assessment of enzyme-negative patients with precordial chest pain of recent onset. PMID- 17408607 TI - BH4 responsiveness associated to a PKU mutation with decreased binding affinity for the cofactor. AB - BACKGROUND: Tetrahydrobiopterin (BH4), cofactor of phenylalanine hydroxylase, can be used to treat a subset of phenylketonuria (PKU) patients as it results in a reduction in blood phenylalanine levels. The molecular basis of the response appears to be multifactorial. METHOD: A standard BH4 loading test (20 mg/kg) was performed. Genotyping was performed by DGGE and sequencing analysis. Expression analysis of the D129G mutation was performed in E. coli (expression as fusion protein MBP-PAH) and in a human hepatoma cell line with an N-terminal FLAG epitope. RESULTS: We report the positive response and long-term treatment of a patient functionally hemizygous for the D129G mutation in the phenylalanine hydroxylase gene. Expression in the prokaryotic system revealed partial activity and a decreased binding affinity for BH4 of the mutant protein. In the eukaryotic system the mutant protein shows reduced stability. CONCLUSION: The D129G mutation which confers a BH4-responsive phenotype, has a decreased binding affinity for BH4. PMID- 17408608 TI - Improving the comparability of immunoassays for prostate-specific antigen (PSA): progress and problems. AB - BACKGROUND: Commutability of immunoassay test results is an important objective for laboratory medicine. METHODS: PSA is a clinically important analyte for which, as a consequence of a number of national and international initiatives over the last decade, considerable progress has been made towards improving method comparability. However, results from different assays are still not interchangeable, a situation that is only likely to improve once broad recommendations can be made about the most clinically relevant antibody combinations. CONCLUSIONS: Universal implementation of such recommendations would almost certainly improve between-method agreement substantially, provided careful attention were paid to assay design and use of appropriately pure secondary standards ensured. PMID- 17408609 TI - Measurement of Elecsys NT-proBNP in serum, K2 EDTA and heparin plasma. AB - OBJECTIVE: There is controversial evidence on a matrix influence on the measurement of NT-proBNP. DESIGN AND METHODS: We compared results of Elecsys NT proBNP measurement on serum, K2 EDTA plasma and lithium heparin plasma. RESULTS: Samples collected in K2 EDTA showed a marginally significant underestimation when compared to serum and heparin, whereas no significant difference was observed between serum and heparin plasma. CONCLUSIONS: Serum, heparin and K2 EDTA plasma may be suitable for NT-proBNP measurement. PMID- 17408610 TI - False positives in plasma ammonia measurement and their clinical impact in a pediatric population. AB - OBJECTIVE: Plasma ammonia measurement is greatly influenced by pre-analytical conditions, which may lead to false positives. We wanted to evaluate the prevalence and clinical impact of plasma ammonia false positives in a pediatric population. DESIGN AND METHODS: Over a 28-month period, charts of patients with elevated ammonemia were retrospectively reviewed to identify false positives defined as elevated concentrations that subsequently normalized without plausible explanation for the elevations. RESULTS: 1880 Ammonia measurements were available in 479 patients. Elevated results that subsequently normalized were found in 86 patients. Forty-one (48%) of these patients had most likely falsely elevated ammonemia. Additional blood sampling and laboratory testing were the most frequent consequences of false positives. CONCLUSION: There is a high proportion of false positives among elevated plasma ammonia measurements. Capillary samples and delay between sampling and centrifugation are possible contributing factors. Clinical consequences of false positives were most often limited. PMID- 17408611 TI - Suspended sediment and sediment-associated contaminants in San Francisco Bay. AB - Water-quality managers desire information on the temporal and spatial variability of contaminant concentrations and the magnitudes of watershed and bed-sediment loads in San Francisco Bay. To help provide this information, the Regional Monitoring Program for Trace Substances in the San Francisco Estuary (RMP) takes advantage of the association of many contaminants with sediment particles by continuously measuring suspended-sediment concentration (SSC), which is an accurate, less costly, and more easily measured surrogate for several trace metals and organic contaminants. Continuous time series of SSC are collected at several sites in the Bay. Although semidiurnal and diurnal tidal fluctuations are present, most of the variability of SSC occurs at fortnightly, monthly, and semiannual tidal time scales. A seasonal cycle of sediment inflow, wind-wave resuspension, and winnowing of fine sediment also is observed. SSC and, thus, sediment-associated contaminants tend to be greater in shallower water, at the landward ends of the Bay, and in several localized estuarine turbidity maxima. Although understanding of sediment transport has improved in the first 10 years of the RMP, determining a simple mass budget of sediment or associated contaminants is confounded by uncertainties regarding sediment flux at boundaries, change in bed-sediment storage, and appropriate modeling techniques. Nevertheless, management of sediment-associated contaminants has improved greatly. Better understanding of sediment and sediment-associated contaminants in the Bay is of great interest to evaluate the value of control actions taken and the need for additional controls. PMID- 17408612 TI - Differential effects of Zn2+ on the kinetics and cocaine inhibition of dopamine transport by the human and rat dopamine transporters. AB - Zn2+ may play a major role in the modulation of neurotransmission because it modulates membrane receptors and channels. Recent literature has shown Zn2+ inhibits dopamine transport by the dopamine transporter (DAT), the main target of cocaine and some other drugs of abuse. Cocaine inhibits DAT and modulation of the DAT by Zn2+ may alter effects of cocaine on dopamine neurotransmission. This study investigates how Zn2+ changes DAT kinetics and its inhibition by cocaine. Steady-state and pre-steady-state kinetics of DAT activity were investigated using rotating disk electrode voltammetry. Values of KM and Vmax in hDAT and effects of cocaine match those in the literature. Zn2+ allosterically inhibited transport in the human DAT (hDAT) with a KI=7.9+/-0.42 microM. Removal of endogenous Zn2+ with penicillamine in hDAT increased transport values. In contrast, Zn2+ did not alter transport by rat DAT (rDAT), with KM and Vmax values of 1.2+/-0.49 microM and 15.7+/-2.57 pmol/(sx10(6) cells), respectively, and removal of Zn2+ did not increase dopamine transport values. Zn2+ allosterically reduced the inhibition by cocaine in hDAT. Results of pre-steady-state studies demonstrated that Zn2+ increases the second order binding rate constant for dopamine to hDAT (3.5 fold to 19.2x10(6) M-1 s-1 for hDAT). In rat striatal homogenates Zn2+ increased initial dopamine transport velocity and decreased cocaine inhibition providing evidence for differences in sensitivity to Zn2+ between the three different preparations. Modulation of the DAT by Zn2+ needs to be assessed further in development of cocaine antagonists. PMID- 17408613 TI - Hypophagic effect of the angiotensin AT1 receptor antagonist irbesartan in rats. AB - Recent experimental and clinical studies report beneficial metabolic effects of antihypertensive drugs interfering with angiotensin. Antagonists at the angiotensin AT(1) receptor can reduce blood glucose and triglyceride levels. So far, there is little evidence, however, that angiotensin AT(1) receptor antagonists can also affect food intake. Particularly unknown is if drugs of this class can have acute effects on short term feeding. To address this issue, the angiotensin AT(1) receptor antagonist irbesartan was studied in a one-hour feeding paradigm in rats. In this study, irbesartan was investigated in comparison with fenfluramine, an established satiating drug, and the angiotensin converting enzyme (ACE) inhibitor captopril. We found a significant reduction of one-hour food intake following 100-200 mg/kg (i.p.) irbesartan. The ACE inhibitor captopril (25-100 mg/kg i.p.) remained without effect on food intake and fenfluramine showed the expected hypophagic action starting at 1 mg/kg (i.p.). The hypophagic effect of irbesartan could not be attributed to sedation or any gross effect on motor activity as determined both upon feeding and independent activity experiments. Fenfluramine (1 mg/kg) and irbesartan (100 mg/kg) did not reduce the latency to feed, but similarly reduced the eating rate at the beginning of the test meal. In conclusion, the present study demonstrates a hypophagic effect of the angiotensin AT(1) receptor antagonist irbesartan that cannot be attributed to sedation or antidipsic effects of the drug. PMID- 17408614 TI - Reduction in methamphetamine induced sensitization and reinstatement after combined pergolide plus ondansetron treatment during withdrawal. AB - We have previously found the 5-HT3 receptor antagonist ondansetron to be useful in reducing cocaine self-administration and cocaine induced sensitization in rats when given during cocaine withdrawal. More recently we have found the combination of the dopamine agonist pergolide plus ondansetron, 3.5 h later, to reverse cocaine sensitization and associated changes in NMDA and AMPA receptors. Here we tested this drug combination in 1) a methamphetamine sensitization model and 2) a reinstatement model after intravenous methamphetamine self-administration using a nose-poke task. We found pergolide plus ondansetron given from days 3-7 of methamphetamine withdrawal to reverse methamphetamine induced sensitization and attenuate reinstatement. We hypothesize that pergolide may evoke a methamphetamine associated memory and that ondansetron can disrupt its reconsolidation. These data suggest that pergolide plus ondansetron treatment may be useful as a therapy to reduce relapse in methamphetamine abusers. PMID- 17408615 TI - Fis1, DLP1, and Pex11p coordinately regulate peroxisome morphogenesis. AB - Dynamin-like protein 1 (DLP1) and Pex11pbeta function in morphogenesis of peroxisomes. In the present work, we investigated whether Fis1 is involved in fission of peroxisomes. Endogenous Fis1 was morphologically detected in peroxisomes as well as mitochondria in wild-type CHO-K1 and DLP1-defective ZP121 cells. Subcellular fractionation studies also revealed the presence of Fis1 in peroxisomes. Peroxisomal Fis1 showed the same topology, i.e., C-tail anchored membrane protein, as the mitochondrial one. Furthermore, ectopic expression of FIS1 induced peroxisome proliferation in CHO-K1 cells, while the interference of FIS1 RNA resulted in tubulation of peroxisomes, hence reducing the number of peroxisomes. Fis1 interacted with Pex11pbeta, by direct binding apparently involving the C-terminal region of Pex11pbeta in the interaction. Pex11pbeta also interacted with each other, whereas the binding of Pex11pbeta to DLP1 was not detectable. Moreover, ternary complexes comprising Fis1, Pex11pbeta, and DLP1 were detected by chemical cross-linking. We also showed that the highly conserved N-terminal domain of Pex11pbeta was required for the homo-oligomerization of Pex11pbeta and indispensable for the peroxisome-proliferating activity. Taken together, these findings indicate that Fis1 plays important roles in peroxisome division and maintenance of peroxisome morphology in mammalian cells, possibly in a concerted manner with Pex11pbeta and DLP1. PMID- 17408616 TI - Influence of triamcinolone intravitreal injection on retinochoroidal healing processes. AB - To analyze the effects of triamcinolone intravitreal injection on the wound healing processes after argon laser retinal photocoagulation, wild type C57BL/6J mice, 8-12 weeks old underwent a standard argon laser photocoagulation protocol. After pentobarbital anesthesia and pupil dilatation, argon laser lesions were induced (50microm, 400mW, 0.05s). Two photocoagulation impacts created two disc diameters from the optic nerve in both eyes. The photocoagulated mice were divided into four groups: Group I (n=12), photocoagulation controls, did not receive any intravitreous injection. Group II (n=12), received an intravitreous injection of 1microl of balanced salt solution (BSS). Group III (n=12), received an intravitreous injection of 1microl containing 15microg of triamcinolone acetonide (TAAC) in BSS. Two mice from each of these three groups were sacrificed at 1, 3, 7, 14 days and 2 and 4 months after photocoagulation. Group IV (n=10) received 1.5, 3, 7.5, 15, or 30microg of TAAC and were all sacrificed on day 14. The enucleated eyes were subjected to systematic analysis of the cellular remodeling processes taking place within the laser lesion and its vicinity. To this purpose, specific antibodies against GFAP, von Willebrand factor, F4/80 and KI67 were used for the detection of astrocytes, activated Muller cells, vascular endothelial cells, infiltrating inflammatory cells and actively proliferating cells. TUNEL reaction was also carried out along with nuclear DAPI staining. Temporal and spatial observations of the created photocoagulation lesions demonstrate that 24h following the argon laser beam, a localized and well delineated affection of the RPE cells and choroid is observed in mice in Groups I and II. The inner retinal layers in these mice eyes are preserved while TUNEL positive (apoptotic) cells are observed at the retinal outer nuclear layer level. At this stage, intense staining with GFAP is associated with activated retinal astrocytes and Muller cells throughout the laser path. From day 3 after photocoagulation, dilated new choroidal capillaries are detected on the edges of the laser lesion. These processes are accompanied by infiltration of inflammatory cells and the presence of proliferating cells within the lesion site. Mice in Group III treated with 15microg/mul of triamcinolone showed a decreased number of infiltrating inflammatory cells and proliferating cells, which was not statistically significant compared to uninjected laser treated controls. The development of new choroidal capillaries on the edges of the laser lesion was also inhibited during the first 2 months after photocoagulation. However, on month 4 the growth of new vessels was observed in these mice treated with TAAC. Mice of Group IV did not show any development of new capillaries even with small doses. After argon laser photocoagulation of the mouse eye, intravitreal injection of triamcinolone markedly influenced the retina and choroid remodeling and healing processes. Triamcinolone is a powerful inhibitor of the formation of neovessels in this model. However, this inhibition is transient. These observations should provide a practical insight for the mode of TAAC use in patients with wet AMD. PMID- 17408617 TI - Temporal response properties of the primary and secondary rod-signaling pathways in normal and Gnat2 mutant mice. AB - Multiple signaling pathways have been proposed for rod vision in the mammalian retina. The primary and secondary rod pathways have been characterized in humans with the scotopic 15-Hz flicker electroretinogram (ERG). The purpose of this study was to determine whether the response properties of these pathways in the mouse are similar to those of humans. C57BL/6J and Gnat2(cpfl3) mutant mice lacking functional cones were used in these experiments. Standard ERG recording techniques were employed. Response functions were obtained for a range of flash intensities (-4.7logcd-s/m(2) to -0.2logcd-s/m(2)) and temporal modulation frequencies (1-30Hz). The mouse intensity-response functions to 15-Hz flickering stimuli possessed the same features as that of humans - a local amplitude minimum and a rapid phase change in the intensity region where the primary and secondary pathways are mutually inhibitory. However, the secondary pathway in the mouse did not achieve the same level of sensitivity as previously shown for humans, suggesting inter-species differences in post-receptoral signal processing. In Gnat2(cpfl3) mutant mice, the secondary pathway was completely abolished. Measurements of temporal acuity indicated that the primary and secondary rod pathways could mediate temporal frequencies as high as 30 and 50Hz, respectively. The response functions for mice are similar to those of humans, although the evidence suggests that the primary rod pathway dominates all rod-mediated signal processing in the mouse. Nevertheless, these results demonstrate the feasibility of measuring non-invasively the performance characteristics of the primary and secondary rod retinal pathways in the mouse and provide a mechanism for testing hypotheses about the action of disease where post-receptoral cells are differentially affected. PMID- 17408618 TI - A critical appraisal of the NXY-059 neuroprotection studies for acute stroke: a need for more rigorous testing of neuroprotective agents in animal models of stroke. AB - Neuroprotection represents a failed strategy to improve outcome after acute ischemic stroke (AIS). However, most neuroprotective drugs have been inadequately studied in animal stroke models, which led to the creation of the STAIR guidelines on preclinical and clinical testing of therapeutics for AIS. NXY-059, a free radical spin trap agent, was felt by many to have followed these criteria and it was recently shown to improve outcome in AIS patients in the SAINT I trial. However, the repeat, SAINT II trial was a neutral study, the results of which cast doubt on neuroprotection as a viable strategy for AIS. A critical analysis of the NXY-059 preclinical data, however, reveals several shortcomings that have not been addressed in the literature. This report contends that the preclinical evaluation of NXY-059 lacked strenuous testing and was not shown to reproducibly lead to robust protection in extended time windows in clinically relevant stroke models, at several different academic research laboratories. The clinical trials of NXY-059 were inadequately designed, in part, because of inappropriate treatment windows and inclusion of diverse stroke patients. Future neuroprotective agents need more rigorous testing in animal models of focal cerebral ischemia and appropriate evaluation in clinical studies that better match the preclinical data. PMID- 17408619 TI - Phytochelatins are synthesized by two vacuolar serine carboxypeptidases in Saccharomyces cerevisiae. AB - Phytochelatins (PCs) are cysteine-rich peptides that chelate heavy metal ions, thereby mediating heavy metal tolerance in plants, fission yeast, and Caenorhabditis elegans. They are synthesized from glutathione by PC synthase, a specific dipeptidyltransferase. While Saccharomyces cerevisiae synthesizes PCs upon exposure to heavy metal ions, the S. cerevisiae genome does not encode a PC synthase homologue. How PCs are synthesized in yeast is unclear. This study shows that the vacuolar serine carboxypeptidases CPY and CPC are responsible for PC synthesis in yeast. The finding of a PCS-like activity of these enzymes in vivo discloses another route for PC biosynthesis in eukaryotes. PMID- 17408620 TI - Expression of ATP binding cassette-transporter ABCG1 prevents cell death by transporting cytotoxic 7beta-hydroxycholesterol. AB - Oxysterols result from cholesterol by enzymatic or oxidative processes. Some exert cytotoxic effects leading to necrosis or apoptosis. Detoxification of these compounds mainly occurs in the liver and requires transport from peripheral tissues towards it. Some ATP-binding cassette transporters are involved in export of cytotoxic compounds. In the current study, we investigated whether ABC transporter family member G1 (ABCG1) may be involved in oxysterol transport, since its gene expression is highly responsive to oxysterol loading. TetOff HeLa cells stably expressing ABCG1 showed decreased mass uptake of 7beta hydroxycholesterol (7beta-HC) whereas that of other physiologically relevant oxysterols was unaffected. Application of 7beta-HC to ABCG1 expressing cells induced hyperpolarization of mitochondrial membrane potential and production of reactive oxygen species, indicating energy consumption by the ATP-binding cassette transporter when it is activated by its correct substrate. Our study points to detoxification as one of potential cellular functions of ABCG1. We assume that ABCG1 protects against 7beta-HC-induced cell death, an important role in prevention of neurodegenerative and cardiovascular disease. PMID- 17408621 TI - Cyclin B1/Cdk1 binds and phosphorylates Filamin A and regulates its ability to cross-link actin. AB - Substantial actin remodelling occurs prior to mitosis as cells alter their shape in preparation for cytokinesis. In mammalian cells, mitosis is initiated by a heterodimer of cyclin B1 and the cyclin dependent kinase 1 (Cdk1) serine/threonine kinase. In this report. we show that human cyclin B1 binds the actin cross-linking protein Filamin-A (FLNa). The proteins co-immunoprecipitate and co-localize in mitotic human cells. We find that cyclin B1/Cdk1 can phosphorylate FLNa in vitro and reduce its ability to gelate actin. We have also identified serine 1436 as one FLNa residue phosphorylated by cyclin B1/Cdk1 in vitro. Our results suggest a role for cyclin B1/Cdk1 in FLNa-dependent actin remodelling. PMID- 17408622 TI - Laparoscopically assisted creation of a neovagina using the Louisville modification. AB - OBJECTIVE: To report a modification of the laparoscopic Davydov method for creation of a neovagina. DESIGN: Case study. SETTING: University gynecology practice. PATIENT(S): Female with vaginal agenesis. INTERVENTION(S): Laparoscopically assisted creation of a neovagina using a modified technique to improve both structural and functional capacity. MAIN OUTCOME MEASURE(S): Patient morbidity and satisfaction. RESULT(S): The patient satisfactorily underwent creation of a neovagina using the laparoscopic modification of the Davydov technique via the Louisville method. CONCLUSION(S): This laparoscopic modification allows for structural maintenance and functional capacity without increasing postoperative morbidity. PMID- 17408623 TI - Incidental testicular cancers that subsequently developed in oligozoospermic and azoospermic patients: report of three cases. AB - OBJECTIVE: To examine whether infertile men with poor semen count subsequently developed testicular cancers and to describe their clinical presentation. DESIGN: We reviewed 460 male patients with abnormal semen counts between 1989 and 2004. SETTING: University hospital. PATIENT(S): Infertile men who developed testicular cancers after assisted reproductive technologies (ART). INTERVENTION(S): Description of patient characteristics: age at infertility, presentation, semen quality, and ART. MAIN OUTCOME MEASURE(S): The number of patients who subsequently developed testicular cancers and the period from ART to the development of clinical testicular cancers. RESULT(S): Of the 460 patients, 169 patients presented with mild oligozoospermia, 117 patients with severe oligozoospermia, and 174 patients with azoospermia. The follow-up periods were as follows: 1-192 months (median, 96.5 mo) for mild oligozoospermia, 1-156 months (median, 78.5 mo) for severe oligozoospermia, and 1-197 months (median, 99 mo) for azoospermia. We subsequently found three testicular cancers that had developed among severely oligozoospermic and azoospermic patients. The period from the claim of sterility to developing testicular cancers varied from 4 to 14 years (median, 9 y). CONCLUSION(S): These results indicate that severe semen abnormality may be a risk factor in developing testicular cancers. Self examination of the testes could be used as an alternative or supplement to physical examination and testicular ultrasound as part of the infertility workup, even after ART. PMID- 17408624 TI - Clinical outcome of cystectomy compared with unilateral salpingo-oophorectomy as fertility-sparing treatment of borderline ovarian tumors. AB - OBJECTIVE: To compare recurrence rates and fertility outcomes of patients with borderline ovarian tumors (BOTs) who underwent unilateral salpingo-oophorectomy with those of patients who underwent cystectomy only. DESIGN: Retrospective study. SETTING: Gynecologic oncology department of a tertiary center. PATIENT(S): Sixty-two patients with BOTs who underwent fertility-preserving surgery. INTERVENTION(S): Unilateral salpingo-oophorectomy (USO, n = 40) or cystectomy only (n = 22). MAIN OUTCOME MEASURE(S): Tumor recurrence rate, incidence of pregnancy. RESULT(S): All 62 patients were alive with no clinical evidence of disease after a mean follow-up of 88 months. There was no statistically significant difference in mean tumor recurrence rates between patients who had undergone cystectomy only and those who had undergone USO (22.7% and 27.5%, respectively). In the cystectomy-treated group, the disease-free interval was shortened (23.6 compared with 41 mo), but the difference was not significant. However, the mean follow-up period for the cystectomy group was significantly shorter than for the USO group. Of the 62 patients, 25 (40.3%) attained 38 pregnancies, resulting in 35 deliveries. CONCLUSION(S): Our results support previous findings that conservative surgery is an acceptable option for women with BOTs who wish to preserve fertility. Cystectomy, like oophorectomy, appears to be an adequate treatment, provided that the patient is willing to undergo careful and prolonged follow-up. PMID- 17408625 TI - External validation of a prediction model for an ongoing pregnancy after intrauterine insemination. AB - OBJECTIVE: To assess the accuracy of our recently developed prediction model in a prospective validation study to predict the outcome of intrauterine insemination (IUI). DESIGN: Descriptive prospective validation study. SETTING: Seven fertility centers in the Netherlands. PATIENT(S): Couples treated with IUI of whom the female partner had a regular cycle. INTERVENTION(S): Intrauterine insemination with or without controlled ovarian hyperstimulation. MAIN OUTCOME MEASURE(S): Ongoing pregnancy after intrauterine insemination. Performance of the prediction model was assessed with calibration and discriminative capacity. Calibration was assessed by comparing the predicted ongoing pregnancy rate with the observed ongoing pregnancy rate. Discriminative capacity was assessed with receiver operation characteristic (ROC) analysis. For daily practice, a score worksheet of the validated model was developed to estimate the chance of an ongoing pregnancy after IUI per treatment cycle. RESULT(S): We included 1,079 subfertile couples who underwent 4,244 cycles of IUI. There were 278 ongoing pregnancies, that is, an ongoing pregnancy rate of 6.6% per cycle. External validation of the model showed good calibration. The predicted probability never differed by more than 1.5% of the mean observed probability. The area under the ROC curve was 0.56 (95% confidence interval, 0.53-0.59) at external validation. CONCLUSION(S): The prediction model was able to make a good distinction between couples with a good pregnancy chance and those with a poor pregnancy chance after IUI. This model can help in deciding which couples will benefit from IUI and which couples will not. PMID- 17408626 TI - Ghrelin in vitro modulates vasoactive factors in human umbilical vein endothelial cells. AB - OBJECTIVE: To determine whether Ghrelin could affect prostaglandins (PGs) and nitric oxide synthesis in human umbilical vein endothelial cells (HUVEC). The effect of Ghrelin on endothelial cell proliferation was also evaluated. DESIGN: In vitro research report. SETTING: Third-level referral academic centers, including molecular and cellular biology laboratories. PATIENT(S): Human umbilical cords were obtained from healthy female volunteers at term of uncomplicated pregnancies. INTERVENTION(S): HUVEC were cultured with Ghrelin (from 10(-11) to 10(-7) M). After 24 hours supernatants were collected and HUVEC were treated for total RNA extraction. MAIN OUTCOME MEASURE(S): In the culture medium PGs release was evaluated by RIA. Prostaglandin-endoperoxide synthase 2 (COX2) and both the constitutive and the inducible isoforms of nitric oxide synthases (ECNOS and INOS) mRNA expressions were evaluated by retrotranscriptase polymerase chain reaction. Endothelial cell proliferation was evaluated by bromo deoxy-uridine incorporation and by cell counting. RESULT(S): Ghrelin negatively affected PGs release as well as COX2, ECNOS, and INOS mRNA expressions in HUVEC. Furthermore, Ghrelin increased bromo-deoxy-uridine incorporation in HUVEC without affecting cell counting. CONCLUSION(S): Our in vitro results allowed to hypothesize that Ghrelin could be involved in the modulation of vascular tone by affecting nitric oxide-related protein synthesis and PGs production in endothelial cells. PMID- 17408627 TI - Ejaculatory-duct calculus causing secondary obstruction and infertility. AB - OBJECTIVE: To report on a patient with ejaculatory-duct calculi that caused secondary obstruction and infertility. DESIGN: Case report. SETTING: Clinical. PATIENT: A 32-year-old man. INTERVENTIONS: Magnetic resonance imaging, vasography, transurethral resection of the veru, and a review of similar cases. RESULT(S): A 32-year-old man with ejaculatory-duct obstruction because of calculi that caused infertility is described. Diagnosis was via vasography and magnetic resonance imaging. Management of this condition involves transurethral veru resection. CONCLUSIONS: Calculus causing ejaculatory-duct obstruction and leading to infertility is a rarely reported phenomenon. Transurethral resection of the veru and calculi extraction is the optimal management. PMID- 17408628 TI - Effects of testosterone and estrogen treatment on lipolysis signaling pathways in subcutaneous adipose tissue of postmenopausal women. AB - OBJECTIVE: The aim of this study was to investigate the treatment effects of testosterone and estrogen on the expression of proteins and genes involved in adipocyte signal transduction to lipolysis in abdominal subcutaneous adipose tissue of postmenopausal women. DESIGN: An open, randomized clinical study with parallel group comparison. SETTING: Women's health clinical research unit and a research laboratory at a university hospital. PATIENT(S): Thirty-six healthy naturally postmenopausal women. INTERVENTION(S): The participants were randomly given testosterone undecanoate (40 mg every second day) or estradiol valerate (2 mg daily) for 3 months. MAIN OUTCOME MEASURE(S): Expression of proteins and genes involved in adipocyte signal transduction to lipolysis in abdominal subcutaneous adipose tissue, determined by quantitative real-time polymerase chain reaction and Western blot, respectively, and related to plasma glycerol before or during a euglycemic hyperinsulinemic clamp. RESULT(S): Testosterone treatment decreased the expression of hormone-sensitive lipase and increased the expression of phosphodiesterase-3B, whereas no effect of estrogen was observed. Testosterone induced changes in hormone-sensitive lipase expression correlated positively with corresponding changes in basal or clamp-induced plasma glycerol concentrations. CONCLUSION(S): Treatment with testosterone in postmenopausal women down-regulates hormone-sensitive lipase and up-regulates phosphodiesterase-3B expressions in abdominal subcutaneous adipose tissue in relation to changes in vivo of lipolytic activity, which may promote the accumulation of fat. PMID- 17408629 TI - DNA hypermethylation of MAL: a promising diagnostic biomarker for colorectal tumors. PMID- 17408630 TI - The forkhead transcription factor FoxO1 regulates proliferation and transdifferentiation of hepatic stellate cells. AB - BACKGROUND & AIMS: The Forkhead box gene, group O (FoxO) family of Forkhead transcription factors is phopsphorylated and inactivated by the phosphatidylinositol 3-kinase (PI3K)/AKT pathway and regulates a variety of cellular functions. Hepatic stellate cells (HSCs) play a crucial role in liver fibrosis. A fibrotic stimulus causes HSCs to transdifferentiate from a quiescent phenotype to a collagen-producing myofibroblast-like phenotype and to proliferate. METHODS: Mutation/deletion mutants of FoxO1 were introduced into primary rat, mouse, and immortalized human HSCs and assessed for activation, proliferation, and signal transduction. The role of FoxO1 in experimental liver fibrosis was assessed in FoxO1(+/-) and FoxO1(+/+) mice. RESULTS: Platelet derived growth factor (PDGF) or insulin phosphorylates FoxO1 and induces FoxO1 translocation from the nuclei to the cytosol via the PI3K/AKT pathway in HSCs. Constitutively active FoxO1 inhibits proliferation via cell cycle arrest at the G1 phase, whereas dominant-negative FoxO1 enhances proliferation of HSCs even in the presence of the PI3K inhibitor LY294002. In addition, the phosphorylation of FoxO1 is increased during transdifferentiation of HSCs. The transdifferentiation is also inhibited by constitutively active FoxO1 and is accelerated by dominant negative FoxO1. FoxO1 directly induces the expression of p27(kip1) and manganese superoxide dismutase (MnSOD). After bile duct ligation for 3 weeks, FoxO1(+/-) mice are more susceptible to liver fibrosis, consistent with our in vitro results. CONCLUSIONS: FoxO1 plays a crucial role in the transdifferentiation and proliferation of HSCs in liver fibrosis. Hyperinsulinemia inactivates FoxO1 in HSCs, resulting in HSC activation and may result in the fibrosis in nonalcoholic fatty liver disease. PMID- 17408631 TI - Precancerous lesions upon sporadic activation of beta-catenin in mice. AB - BACKGROUND & AIMS: Inappropriate activation of beta-catenin in adult tissues is associated with a wide variety of cancers, especially in the digestive tract. Classic transgenic and knockout murine models in which beta-catenin is activated in large fields of cells have provided experimental support in favor of a role for this molecule in tumorigenesis. However, these models do not reproduce the sporadic nature of the majority of human cancers, beginning with the activation of an oncogene at random in a single cell. METHODS: We used the "hit and run" strategy to generate a mouse model in which the expression of an activated form of beta-catenin occurs sporadically in vivo. RESULTS: Sporadic, multifocal lesions were observed in the stomach of 3% of mice aged 8 months and older. These lesions were associated with loss of Sonic hedgehog (Shh), and a causal relationship between beta-catenin activation and Shh inhibition was established in gastric cells in vitro. No lesion was detected in the intestine or in the liver. In addition, one third of female mutant mice developed benign perimammary papillomas. Mutant mice were also hypersensitive to chemically induced premalignant skin lesions. CONCLUSIONS: These results challenge the view that activation of beta-catenin induces malignant cancerogenesis, because they show in mice that sporadically activated beta-catenin is sufficient for tumor initiation, yet without further malignant progression, and that it sensitizes cells to environmental hits. This model represents a powerful tool to investigate the interplay between genetic and environmental factors in tumor progression. PMID- 17408632 TI - Alcohol-induced protein kinase Calpha phosphorylation of Munc18c in carbachol stimulated acini causes basolateral exocytosis. AB - BACKGROUND & AIMS: Acute or chronic alcohol treatment does little to the exocrine pancreas but predisposes the pancreas to postprandial cholinergic stimulation that triggers cellular events leading to pancreatitis. This alcohol-induced susceptibility mechanism of pancreatitis is unknown. METHODS: We employed alcohol treated dispersed rat pancreatic acini and alcohol diet-fed rats to examine the effects of submaximal carbachol-induced changes in exocytosis (FM1-43 epifluorescence imaging and electron microscopy), Munc18c cellular translocation (confocal microscopy and subcellular fractionation), and protein kinase C (PKC) alpha-induced phosphorylation in relation to pancreatitis. RESULTS: Acute low dose alcohol (20 mmol/L) in vitro exposure or chronic alcohol diet reduces postprandial cholinergic-stimulated amylase secretion from rat pancreatic acinar cells by blocking apical exocytosis and redirecting exocytosis to less efficient basolateral plasma membrane sites. This ectopic exocytosis is mediated by PKCalpha-induced phosphorylation of Munc18c, causing Munc18c displacement from the basolateral plasma membrane into the cytosol in which it undergoes proteolytic degradation; these processes can be blocked by PKCalpha inhibition. CONCLUSIONS: We conclude that sequential low-dose alcohol and postprandial cholinergic stimulation can induce PKCalpha-mediated Munc18c plasma membrane displacement. This relieves cognate SNARE proteins on zymogen granules and basolateral membrane to complex and consummate pathologic ectopic exocytosis at the basolateral surface. This change in vesicle trafficking may be related to the pathogenesis of pancreatitis. PMID- 17408633 TI - Rare CpG island methylator phenotype in ulcerative colitis-associated neoplasias. AB - BACKGROUND & AIMS: We previously reported that a high degree of age-related methylation was found in both the dysplastic and nondysplastic mucosa of patients with ulcerative colitis (UC). Whether this translates into hypermethylation in UC associated cancers (UC-Cs) is not known. METHODS: We evaluated the methylation status of 11 genes (MINT1, 2, 31, hMLH1, p16, p14, MGMT, HPP1, SFRP1, ERalpha, and LINE-1) in 48 UC-Cs, 21 UC-associated dysplasias, and 69 sporadic colorectal cancers (S-CRCs) using a quantitative bisulfite pyrosequencing analysis. RESULTS: Methylation levels in UC-Cs were lower than S-CRCs for all the genes except MGMT. A methylation index based on the average of Z-scores, for type C (cancer-specific genes: MINT1, MINT2, MINT31, hMLH1, p16, and p14) was -.97 in UC-Cs and .92 in S CRCs (P = .009). That of type A (age-related genes: HPP1, SFRP1, and ERalpha) was -1.97 in UC-Cs and 1.24 in S-CRCs (P < .001). We observed a significant difference in the incidence of CpG island methylator phenotype between UC-Cs and S-CRCs (8 of 48 [17%] and 26 of 69 [38%]; P = .022). UC-associated dysplasias had significantly higher methylation of type A gene than UC-Cs (Z-score: .07 and 1.97, respectively; P < .001). By contrast, global DNA methylation measured using a LINE-1 assay was significantly higher in UC-Cs than in S-CRCs (58.2% vs 51.0%, P < .001). CONCLUSIONS: DNA methylation alterations are uncommon in UC cancers. Given that both genetic and epigenetic changes are common in UC mucosa and dysplasias, we speculate that the genetic changes lead to a more aggressive clinical course than epigenetic changes. PMID- 17408634 TI - Does gender affect career satisfaction and advancement in gastroenterology? Results of an AGA institute-sponsored survey. AB - BACKGROUND & AIMS: Women comprise 19% of the American Gastroenterological Association (AGA) membership. We performed a prospective study to determine whether female gastroenterologists were less likely to achieve career advancement and satisfaction. METHODS: We administered an online survey to AGA members from 2004-2006. The survey contained questions regarding effects of gender on career advancement, satisfaction with career, promotional policies, and integration of family and career. RESULTS: A total of 457 individuals (response rate 9% after 2 major invitations) completed the survey, including 262 (57%) women (20% in private practice, 53% in academic careers, and 27% trainees) and 195 men (23% in private practice, 58% in academic careers, and 19% trainees). The male gastroenterologists were significantly older (P < .005) and in their careers for significantly more years (P = .002). There were no significant differences with respect to marital status, number of children, or number of hours worked between the genders. Men were more likely to achieve the rank of full professor (P = .035), and significantly more women reported that gender affected their career advancement (47% vs 9%; P < .001). Women in academic careers reported less satisfaction with their careers (P = .01) and perceived more difficulty in achieving promotion and tenure. Women were more likely to choose private practice careers because of part-time options (P = .025). Equal numbers of men and women in practice reported difficulty balancing work and family life. CONCLUSIONS: Significantly more female than male gastroenterologists perceive that gender has affected their career advancement. Female academic gastroenterologists reported less overall career satisfaction and promotion than male academic gastroenterologists. PMID- 17408635 TI - H-ras inhibits RhoA/ROCK leading to a decrease in the basal tone in the internal anal sphincter. AB - BACKGROUND & AIMS: The present studies evaluated the role of H-ras and its implications in the RhoA/Rho kinase (ROCK) pathway in regulating basal tone in the internal anal sphincter (IAS). METHODS: Studies were performed in the IAS from the wild-type (H-ras(+/+)) and knock-out (H-ras(-/-)) mice. The basal tone of smooth muscle strips was measured by isometric force transducers. Length of smooth muscle cells (SMC) isolated from the IAS in the basal state was determined by phase contrast microscopy. Experiments were repeated in the presence of Y 27632, a ROCK inhibitor. Involvement of the RhoA/ROCK machinery was analyzed by reverse-transcription polymerase chain reaction, Western blot, and immunocytochemistry. Reversal of H-ras knock-out effect was evaluated by transfection of SMCs with the constitutively activated (G12V) mutant. RESULTS: Basal tone of the H-ras(-/-) IAS was significantly higher and resistant to relaxation by Y 27632, compared with the H-ras(+/+) IAS. Similarly, the length of SMCs from H-ras(-/-) IAS was significantly shorter. Y 27632 eliminated this difference. RhoA immunoreactivity shifted from cytoplasm to plasma membrane in H ras(-/-) SMCs, a change typically associated with contraction. Further, SMCs from H-ras(-/-) mice exhibited higher levels of the contractile proteins ROCK II, phosphorylated-MYPT(1) and phosphorylated-MLC(20). Transfection with the G12V mutant increased the length of H-ras(-/-) cells. Conversely, the dominant negative H-ras (S17N) mutant decreased the length of H-ras(+/+) cells. CONCLUSIONS: H-ras negatively regulates basal tone in the IAS by inhibiting RhoA/Rho-kinase machinery. Studies may have significant relevance in the pathophysiology and therapy of certain anorectal motility disorders associated with the IAS dysfunction. PMID- 17408636 TI - Improving colorectal adenoma detection: technology or technique? PMID- 17408637 TI - A novel role of VIP in colonic motility function: induction of excitation transcription coupling in smooth muscle cells. AB - BACKGROUND & AIMS: Vasoactive intestinal polypeptide (VIP) relaxes smooth muscle by generation of cAMP and activation of protein kinase A (PKA). However, PKA activation also phosphorylates the transcription factor CREB. The aim of this study was to investigate whether the phosphorylation of CREB induces gene expression of the pore-forming alpha(1C) subunit of Ca(v)1.2 channels (L-type calcium channels), whose promoter has 2 binding sites for CREB. METHODS: The experiments were performed on primary cultures of human colonic circular smooth muscle cells and freshly obtained human and rat colonic circular muscle strips. RESULTS: The incubation of human colonic circular smooth muscle cells or muscle strips with VIP for 24 hours enhanced the expression of alpha(1C) protein and mRNA as well as the contractile response to acetylcholine and KCl. On the contrary, incubation of the muscle strips with VIP antagonist for 24 hours suppressed cell contractility. The incubation of the cells with VIP caused sustained generation of cAMP for 24 hours, but PKA activation and CREB phosphorylation were transient. The inhibition of PKA by H-89 or silencing of CREB gene with targeted RNAi blocked the transcription of alpha(1C). Progressive 5' deletions of halpha(1C)1b promoter and site-directed mutations of the 2 CREB binding cis-elements indicated that most of alpha(1C) transcription was mediated by the 5' cAMP response element. CONCLUSIONS: The excitation-transcription coupling stimulated by VIP induces expression of the Ca(v)1.2 channels. The influx of calcium through these channels is a critical step in excitation contraction coupling in smooth muscle cells. PMID- 17408638 TI - The forkhead box M1 transcription factor contributes to the development and growth of mouse colorectal cancer. AB - BACKGROUND & AIMS: In this study, we used Forkhead Box m1b (Foxm1b) transgenic mice and conditional Foxm1 knock-out mice to examine the role of Foxm1 in colon cancer development and proliferation. METHODS: To induce mouse colorectal cancer, we used a single intraperitoneal injection of azoxymethane (AOM) followed by three 1-week cycles of 2.5% dextran sodium sulfate (DSS) water, each cycle separated by 2 weeks. For these colon tumor studies, we used either Rosa26-Foxm1b transgenic mice that ubiquitously expressed the human Foxm1b complementary DNA or mice in which the Foxm1 fl/fl targeted allele was deleted in colonic epithelial cells using the gut-specific Villin-Cre recombinase transgene (Villin-Cre). Colorectal tumor number and bromodeoxyuridine labeling were determined in Rosa26 Foxm1b mice, Villin-Cre Foxm1-/-, mice and wild-type mice after 12 weeks of AOM/DDS exposure. We also used Foxm1 small interfering RNA-depleted human DLD1 and mouse CT26 colon cancer cell lines to examine DNA replication and anchorage independent growth. RESULTS: After 12 weeks of treatment with AOM/DSS, Rosa26 Foxm1b transgenic mice showed an increase in the number and size of colorectal tumors compared with wild-type mice. Likewise, a significant reduction in the development and growth of colorectal tumors was found in Villin-Cre Foxm1-/- mice compared with Foxm1 fl/fl mice after AOM/DSS treatment, which was associated with decreased expression of cyclin A2, cyclin B1, survivin, and T-cell factor 4 genes. Moreover, Foxm1-depleted colon cancer cell lines showed reduced DNA replication and anchorage-independent growth. CONCLUSIONS: These studies suggest that Foxm1 is critical for the proliferation and growth of colorectal cancer. PMID- 17408640 TI - Toll-like receptor 2 controls mucosal inflammation by regulating epithelial barrier function. AB - BACKGROUND & AIMS: Toll-like receptors (TLRs) represent a class of transmembrane pattern recognition receptors essential for microbial recognition and control of innate immune responses. Commensal bacteria play an important role in maintaining tolerance and active stability of the intestinal epithelial barrier by suppressing intestinal inflammation, yet the mechanisms of action are unknown. The aim of this study was to determine the functional relevance of TLR2 to control tight junction (TJ)-associated intestinal epithelial barrier integrity to balance mucosal homeostasis against inflammatory stress-induced damage. METHODS: TLR2 ligand (synthetic Pam(3)Cys-SK4 [PCSK])-induced activation of signaling cascades and TJ-associated distribution was assessed by using Western blotting and confocal microscopy combined with functional transfection and inhibitor studies in model intestinal epithelial cell (IEC) lines (IEC-6, Caco-2) or primary IEC cultured short-term ex vivo. DSS colitis was induced by standard protocol in wild-type, TLR2-/-, and MyD88-/- mice. Spontaneous apoptosis was assessed by terminal deoxinucleotidyl-transferase-mediated dUTP-biotin nick end labeling. RESULTS: Data from in vitro and ex vivo models of intestinal epithelial cells revealed that TLR2 stimulation effectively preserves TJ-associated barrier assembly against stress-induced damage through promotion of PI3K/Akt-mediated cell survival via MyD88. Furthermore, in vivo studies underscored that TLR2 mediated TJ regulation critically determines susceptibility to intestinal injury and inflammation. Inflammatory stress in mice deficient of TLR2 or MyD88 induced early TJ-associated disruption interrelated with anti-apoptotic failure of the intestinal epithelial barrier. Oral treatment of colitis with the TLR2 ligand PCSK significantly suppressed mucosal inflammation and apoptosis by efficiently restoring TJ-associated integrity of the intestinal epithelium in vivo. CONCLUSION: TLR2 may provide a target to pharmacologically modulate mucosal injury and intestinal inflammation. PMID- 17408641 TI - Periostin creates a tumor-supportive microenvironment in the pancreas by sustaining fibrogenic stellate cell activity. AB - BACKGROUND & AIMS: Pancreatic cancer creates desmoplasia by stimulating stellate cells (PSCs), thereby influencing tumor aggressiveness. The aim of this study was to analyze the impact of the PSC-specific matrix protein periostin on tumor responses to radiochemotherapy. METHODS: PSCs and cancer cells in primary and metastatic lesions of patients treated with or without neoadjuvant radiochemotherapy were evaluated by immunohistochemistry. Periostin messenger-RNA levels determined by quantitative reverse-transcription polymerase chain reaction were correlated to patient survival. Interactions between PSCs and cancer cells and the effects of periostin in modulating cellular responses under conditions of hypoxia, starvation, and radiochemotherapy were assessed by immunoblotting and by growth, clonogenicity, and invasion assays. RESULTS: Periostin messenger-RNA levels were elevated 42-fold in cancer, and patients with increased expression had a tendency toward shorter survival (19 vs 12 months; P = .14). Stromal cells were the only source of periostin in the pancreas and in metastatic sites. Cancer cell supernatants stimulated periostin secretion from PSCs. Recombinant periostin increased alpha-smooth muscle actin, periostin, collagen-1, fibronectin, and transforming growth factor-beta1 expression while decreasing PSC invasiveness. These effects were reversed by silencing periostin expression and secretion by small interfering RNA transfection. In cancer cells, periostin stimulated growth and conferred resistance to starvation and hypoxia. In addition, the periostin downstream target collagen-1 significantly increased chemoresistance. CONCLUSIONS: Once stimulated by cancer cells, PSCs remain active via an autocrine periostin loop even under radiotherapy and produce excessive extracellular matrix proteins, creating a tumor-supportive microenvironment. Increased periostin expression may therefore reflect a more aggressive tumor phenotype. PMID- 17408643 TI - Navigating the maze of hepatitis B treatments. PMID- 17408644 TI - TGF-betaRII rescues development of small intestinal epithelial cells in Elf3 deficient mice. AB - BACKGROUND & AIMS: ELF3, a member of the ETS transcription factor family, has been shown to transactivate the transforming growth factor beta type II receptor (TGF-betaRII) promoter. Previously we showed that Elf3-null mice have a defect in the small intestine caused by a failure of small intestinal epithelial cells to differentiate and that these cells produced significantly lower levels of Tgf betaRII. To prove that the defect observed in Elf3-null mice resulted from the lack of Elf3-dependent activation of Tgf-betaRII expression, we performed a genetic rescue. METHODS: We generated transgenic mice that express human TGF betaRII specifically in the intestinal epithelium under the control of the mouse A33 antigen promoter. Mice expressing the A33-TGF-betaRII transgene were mated with Elf3(+/-) mice, and double heterozygous offspring harboring both the transgene and one mutant Elf3 allele were intercrossed. RESULTS: The resultant A33-TGF-betaRII transgenic Elf3(-/-) pups displayed normal small intestinal morphology, while the characteristic abnormality was retained in all Elf3(-/-) mice that did not express the transgene. This phenotypic rescue shows for the first time in vivo that a single gene, Elf3, is the critical upstream regulator of Tgf-betaRII in mouse small intestinal epithelium. CONCLUSIONS: This has important implications for our understanding of tissue-specific gene regulation and further strengthens the potential clinical connection between ELF3 and colorectal cancer involving transforming growth factor beta insensitivity. PMID- 17408645 TI - Therapeutic doses of acetaminophen frequently cause elevated aminotransferases in healthy volunteers: is it significant? PMID- 17408646 TI - Serotonin-1A receptor gene HTR1A variation predicts interferon-induced depression in chronic hepatitis C. AB - BACKGROUND & AIMS: Interferon-induced depression is a major complication in antiviral therapy of chronic hepatitis C. Little is known about underlying mechanisms and reliable predictive factors associated with cytokine-induced depressive symptoms. METHODS: In a cohort of 139 hepatitis C-infected outpatients treated with interferon alfa-2b, we investigated the impact of functional gene variants of the cerebral serotonin (5-HT) signalling pathway previously implicated in depression risk. Depression was monitored using the Hospital Anxiety and Depression Scale (HADS). All patients were genotyped for functional variations in the 5-HT(1A) receptor (HTR1A), 5-HT transporter (SLC6A4, 5-HTT), and tryptophan hydoxylase-2 (TPH2). RESULTS: Homozygosity for the HTR1A-1019G variant significantly increased both incidence and severity of interferon-induced depression. Maximum increases in HADS depression scores during antiviral therapy correlated with HTR1A variation (P = .011). Clinically relevant depression was significantly associated with the HTR1A-1019G genotype (P = .017; OR, 2.95). 5 HTT and TPH2 variations did not contribute significantly to the prediction of interferon-induced depression by HTR1A (sensitivity, 35.9%; specificity, 84.0%). CONCLUSIONS: Our findings suggest an impact of allelic variation in 5-HT(1A) receptor expression on the development of interferon alfa-induced depression during antiviral treatment of chronic hepatitis C. Prediction models of interferon-induced depressive symptoms based on HTR1A variation offer a perspective for an antidepressant selective serotonin reuptake inhibitor prophylaxis in patients genetically at risk for interferon-induced depression. PMID- 17408647 TI - Transglutaminase 2 regulates mallory body inclusion formation and injury associated liver enlargement. AB - BACKGROUND & AIMS: Mallory body (MB) inclusions are a characteristic feature of several liver disorders and share similarities with cytoplasmic inclusions observed in neural diseases and myopathies. MBs consist primarily of keratins 8 and 18 (K8/K18), require a K8-greater-than-K18 ratio for their formation, and contain glutamine-lysine cross-links generated by transglutaminase (TG). We hypothesized that protein transamidation is essential for MB formation. METHODS: Because TG2 is the most abundant hepatocyte TG, we tested our hypothesis using TG2(-/-) and their wild-type counterpart mice fed 3,5-diethoxycarbonyl-1,4 dihydrocollidine (DDC), an established MB inducer. Keratin cross-linking was further examined using recombinant proteins or transgenic mice that overexpress K8 or K18. RESULTS: TG2(-/-) livers have markedly reduced TG2 activity as compared with TG2(+/+) livers. The DDC-fed TG2(-/-) mice have dramatic decreases in MB formation and liver hypertrophy response as contrasted with DDC-fed TG2(+/+) mice. Despite similar hepatocellular damage, TG2(-/-) mice had more gallstones, jaundice, and ductal proliferation than wild-type mice. Inhibition of MB formation in TG2(-/-) mice was associated with marked attenuation of ubiquitination and K8-containing protein cross-linking. MB formation and resolution paralleled the generation then disappearance of cross-linked K8, respectively. K8 is a preferential TG2 substrate when compared to K18, as examined in vitro or in DDC-fed transgenic mice that overexpress K8 or K18. CONCLUSIONS: We demonstrate an essential role for TG2 in determining injury mediated liver enlargement and the necessity of K8 and TG2 for generating cross linked keratins and MBs. The role of TG in inclusion formation might extend to nonkeratin intermediate filament protein-related diseases. PMID- 17408648 TI - Alterations in the brain-gut axis underlying visceral chemosensitivity in Nippostrongylus brasiliensis-infected mice. AB - BACKGROUND & AIMS: Visceral hypersensitivity, a hallmark of irritable bowel syndrome, is generally considered to be mechanosensitive in nature and mediated via spinal afferents. Both stress and inflammation are implicated in visceral hypersensitivity, but the underlying molecular mechanisms of visceral hypersensitivity are unknown. METHODS: Mice were infected with Nippostrongylus brasiliensis (Nb) larvae, exposed to environmental stress and the following separate studies performed 3-4 weeks later. Mesenteric afferent nerve activity was recorded in response to either ramp balloon distention (60 mm Hg), or to an intraluminal perfusion of hydrochloric acid (50 mmol/L), or to octreotide administration (2 micromol/L). Intraperitoneal injection of cholera toxin B-488 identified neurons projecting to the abdominal viscera. Fluorescent neurons in dorsal root and nodose ganglia were isolated using laser-capture microdissection. RNA was hybridized to Affymetrix Mouse whole genome arrays for analysis to evaluate the effects of stress and infection. RESULTS: In mice previously infected with Nb, there was no change in intestinal afferent mechanosensitivity, but there was an increase in chemosensitive responses to intraluminal hydrochloric acid when compared with control animals. Gene expression profiles in vagal but not spinal visceral sensory neurons were significantly altered in stressed Nb-infected mice. Decreased afferent responses to somatostatin receptor 2 stimulation correlated with lower expression of vagal somatostatin receptor 2 in stressed Nb-infected mice, confirming a link between molecular data and functional sequelae. CONCLUSIONS: Alterations in the intestinal brain-gut axis, in chemosensitivity but not mechanosensitivity, and through vagal rather than spinal pathways, are implicated in stress-induced postinflammatory visceral hypersensitivity. PMID- 17408649 TI - R-spondin1, a novel intestinotrophic mitogen, ameliorates experimental colitis in mice. AB - BACKGROUND & AIMS: R-spondin 1 (Rspo1) is a novel epithelial mitogen that stimulates the growth of mucosa in both the small and large intestine. METHODS: We investigated the therapeutic potential of Rspo1 in ameliorating experimental colitis induced by dextran sulfate sodium (DSS) or trinitrobenzene sulfonic acid (TNBS) as well as nonsteroidal anti-inflammatory drug-induced colitis in interleukin (IL)-10-deficient mice. RESULTS: Therapeutic administration of recombinant Rspo1 protein reduced the loss of body weight, diarrhea, and rectal bleeding in a mouse model of acute or chronic DSS-induced colitis. Histologic evaluation revealed that Rspo1 improved mucosal integrity in both villus and/or crypt compartments in the small intestine and colon by stimulating crypt cell growth and mucosal regeneration in DSS-treated mice. Moreover, Rspo1 significantly reduced DSS-induced myeloperoxidase activity and inhibited the overproduction of proinflammatory cytokines, including tumor necrosis factor alpha, IL-1alpha, IL-6, interferon-gamma, and granulocyte-macrophage colony stimulating factor, in mouse intestinal tissue, indicating that Rspo1 may reduce DSS-induced inflammation by preserving the mucosal barrier function. Likewise, Rspo1 therapy also alleviated TNBS-induced interstitial inflammation and mucosal erosion in the mouse colon. Furthermore, Rspo1 substantially decreased the histopathologic severity of chronic enterocolitis by repairing crypt epithelium and simultaneously suppressing inflammatory infiltration in piroxicam-exposed IL 10(-/-) mice. Endogenous Rspo1 protein was localized to villus epithelium and crypt Paneth cells in mouse small intestine. CONCLUSIONS: Our results show that Rspo1 may be clinically useful in the therapeutic treatment of inflammatory bowel disease by stimulating crypt cell growth, accelerating mucosal regeneration, and restoring intestinal architecture. PMID- 17408650 TI - Enteric glia regulate intestinal barrier function and inflammation via release of S-nitrosoglutathione. AB - BACKGROUND & AIMS: Barrier functions across epithelia and endothelia are essential for homeostatic tissue regulation. Astroglia interact with cerebral endothelia to maintain the blood-brain barrier. Whether similar interactions between astrocyte-like enteric glia and epithelia regulate intestinal barrier function is not known. METHODS: Fluorescent permeability markers were used to measure intestinal barrier function in vivo after conditional ablation of enteric glia in transgenic mice. Enteric glial cell regulation of epithelial barrier integrity then was modeled in vitro using coculture. Glial-derived barrier inducing factors were characterized using size-exclusion chromatography and mass spectrometry. Epithelial barrier integrity was assessed by transepithelial resistance readings and by quantitative measurement of tight-junction-associated protein expression by quantitative polymerase chain reaction and Western blot. RESULTS: We show that ablation of enteric glial cells in transgenic mice causes intestinal mucosal barrier dysfunction, resulting in inflammation. Glial-derived s-nitrosoglutathione (GSNO) was identified as a potent inducer of mucosal barrier function in vitro and in vivo and of attenuated tissue inflammation after ablation of enteric glia in transgenic mice. GSNO regulation of mucosal barrier function was associated directly with an increased expression of perijunctional F actin and tight-junction-associated proteins zonula occludens-1 and occludin. GSNO also significantly restored mucosal barrier function in colonic biopsy specimens from patients with Crohn's disease, a well-described inflammatory permeability disorder associated with enteric glial-cell disruption. CONCLUSIONS: Enteric glia therefore share the ability of astrocytes to regulate tight-junction integrity, and cellular interactions comparable with those maintaining blood brain barrier function also regulate epithelial permeability at mucosal surfaces. PMID- 17408651 TI - Sodium iodide symporter is expressed at the preneoplastic stages of liver carcinogenesis and in human cholangiocarcinoma. AB - BACKGROUND & AIMS: The ability of thyroid cells to take up iodide, which enables (131)I radiotherapy for thyroid cancer, is due to the expression of the sodium iodide symporter at their plasma membrane. Expression of this symporter has been found in some nonthyroid cancers. However, it is mostly accumulated in the cytoplasm, and its functionality has not been demonstrated. We have investigated sodium iodide symporter expression and functionality in human liver cancer, and in a diethylnitrosamine induced Wistar rat model of primary liver cancer at different stages of carcinogenesis. METHODS: Sodium iodide symporter mRNA and protein were studied in tissues from patients with hepatocellular- or cholangio carcinomas using reverse-transcription polymerase chain reaction, immunoblot, and immunohistochemistry. We studied the dynamics of hepatic iodine uptake in the animal model using nuclear imaging. RESULTS: Sodium iodide symporter expression showed up in all 20 cholangiocarcinomas, but in only 2 of the 26 hepatocellular carcinomas, investigated. It was also found in normal bile duct cells and in the ductular reaction present in cirrhotic tissues. It was located at the plasma membrane in 10 of 20 cholangiocarcinoma. In rat liver cancer, a functional sodium iodide symporter expression was triggered as from the early preneoplastic steps, and was amplified during clonal tumor cell expansion, allowing complete tumor suppression after (131)I radiotherapy. CONCLUSIONS: A significant proportion of human cholangiocarcinomas expresses membrane sodium iodide symporter, which may permit radioiodine therapy. Our data also suggest that (131)I acts on a crucial target for liver cancer development. PMID- 17408652 TI - Risk factors for mortality after surgery in patients with cirrhosis. AB - BACKGROUND & AIMS: Current methods of predicting risk of postoperative mortality in patients with cirrhosis are suboptimal. The utility of the Model for End-stage Liver Disease (MELD) in predicting mortality after surgery other than liver transplantation is unknown. The aim of this study was to determine the risk factors for postoperative mortality in patients with cirrhosis. METHODS: Patients with cirrhosis (N = 772) who underwent major digestive (n = 586), orthopedic (n = 107), or cardiovascular (n = 79) surgery were studied. Control groups of patients with cirrhosis included 303 undergoing minor surgical procedures and 562 ambulatory patients. Univariate and multivariable proportional hazards analyses were used to determine the relationship between risk factors and mortality. RESULTS: Patients undergoing major surgery were at increased risk for mortality up to 90 days postoperatively. By multivariable analysis, only MELD score, American Society of Anesthesiologists class, and age predicted mortality at 30 and 90 days, 1 year, and long-term, independently of type or year of surgery. Emergency surgery was the only independent predictor of duration of hospitalization postoperatively. Thirty-day mortality ranged from 5.7% (MELD score, <8) to more than 50% (MELD score, >20). The relationship between MELD score and mortality persisted throughout the 20-year postoperative period. CONCLUSIONS: MELD score, age, and American Society of Anesthesiologists class can quantify the risk of mortality postoperatively in patients with cirrhosis, independently of the procedure performed. These factors can be used in determining operative mortality risk and whether elective surgical procedures can be delayed until after liver transplantation. PMID- 17408653 TI - Biliary epithelial cell antibodies link adaptive and innate immune responses in primary sclerosing cholangitis. AB - BACKGROUND & AIMS: Primary sclerosing cholangitis (PSC) is an autoimmune liver disease with destruction of hepatic bile ducts. A high frequency of biliary epithelial cell antibodies (BEC-Ab) is present in PSC. Here, we studied the mechanisms and signaling pathways used by these Ab in causing BEC dysfunction. METHODS: Immunoassays were performed using freshly isolated BECs to study the signaling capacity of purified immunoglobulin (Ig) G and F(ab)'(2) fractions from 33 patients with PSC with anti-BEC-Ab. RESULTS: We provide evidence that stimulation of BECs with PSC IgG, but not control IgG, induced expression of Toll like receptor (TLR) 4 and TLR9 and specific phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 as well as the transcription factors ELK-1 and nuclear factor kappaB. A specific inhibitor of ERK1/2 abrogated phosphorylation of ELK-1 and protein expression of TLR4 but not TLR9 on BECs. TLR-expressing BECs, when further stimulated with lipopolysaccharide and CpG DNA, produced high levels of interleukin-1beta, interleukin-8, interferon gamma, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, and transforming growth factor beta. Bile ducts stained positively for TLR4 and TLR9 in 58% of liver specimens taken from patients with PSC with BEC-Ab, as compared with 14% in those without BEC-Ab and also less frequently in diseased control livers. CONCLUSIONS: Our data show that binding of PSC BEC-Ab initiates ERK1/2 signaling and up-regulation of TLR, which upon ligation induces BECs to produce cytokines/chemokines, leading to the possible recruitment of inflammatory cells. Thus, in PSC, BECs are not only targets of the immune attack but may also be active participants and mediators of their own destruction. BEC-Ab may be critical regulators of cholangitis in PSC. PMID- 17408655 TI - Opportunities and Challenges at NIDDK in Digestive Diseases Research. PMID- 17408654 TI - Pain in chronic pancreatitis: the role of reorganization in the central nervous system. AB - BACKGROUND & AIMS: In various chronic pain conditions cortical reorganization seems to play a role in the manifestations. The aim of this study was to investigate cortical reorganization in patients with pain caused by chronic pancreatitis. METHODS: Twelve healthy subjects and 10 patients with chronic pancreatitis were included. The esophagus, stomach, and duodenum were stimulated electrically at the pain threshold using a nasal endoscope. The electroencephalogram was recorded from 64 surface electrodes and event-related brain potentials (EPs) were obtained. RESULTS: As compared with healthy subjects, the patient group showed decreased latencies of the early EP components (N1, P < .001; P1, P = .02), which is thought to reflect the exogenous brain pain processing specifically. Source analysis showed that the dipolar activities corresponding to the early EPs were located consistently in the bilateral insula, in the anterior cingulate gyrus, and in the bilateral secondary somatosensory area. The bilateral insular dipoles were localized more medial in the patient group than in the healthy subjects after stimulation of all 3 gut segments (P < .01). There also were changes in the cingulate cortex where the neuronal source was more posterior in patients than in controls to stimulation of the esophagus (P < .05). CONCLUSIONS: The findings indicate that pain in chronic pancreatitis leads to changes in cortical projections of the nociceptive system. Such findings also have been described in somatic pain disorders, among them neuropathic pain. Taken together with the clinical data this suggests a neuropathic component in pancreatic pain, which may influence the approach to treatment. PMID- 17408657 TI - Enteric neuroscience: the dawn of a new discipline. PMID- 17408658 TI - Drug targets and molecular mechanisms of drug resistance in chronic hepatitis B. AB - Chronic hepatitis B continues to be a major cause of end-stage liver disease and hepatocellular carcinoma worldwide. Nucleos(t)ide analogues have proven to be effective in controlling the disease and perhaps decreasing the incidence of hepatocellular carcinoma. However, development of drug resistance is a major limitation to their long-term effectiveness. Understanding the mechanisms of drug resistance are important for designing new agents and devising strategies to manage and prevent the development of antiviral drug resistance. The development of resistance is determined by an interplay of viral, host, and drug characteristics Homology of the HBV polymerase to the human immunodeficiency virus-1 reverse transcriptase has allowed predictions to be made on the effect mutations have on HBV polymerase structure. In vitro functional studies provide complementary information. Several broad principles on the mechanism of resistance have emerged from these studies. First, most of the primary mutations cluster in the vicinity of the incoming nucleotide and act by directly affecting the position or stability of the bound substrate, template, or primer. In contrast, secondary mutations tend to occur away from the nucleotide-binding pocket. Finally, the structural and functional consequences of mutations are quite variable among the different agents. This paper reviews the key mutations and mechanisms associated with resistance to the nucleos(t)ide analogues approved for clinical use and discuss new targets for drug development. PMID- 17408659 TI - Lectin conjugates as potent, nonabsorbable CFTR inhibitors for reducing intestinal fluid secretion in cholera. AB - BACKGROUND & AIMS: Inhibitors of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel are predicted to prevent intestinal fluid secretion in cholera. We previously discovered low- affinity glycine hydrazide (GlyH) CFTR inhibitors that block CFTR at its external pore. The goal of this study was to develop potent CFTR inhibitors that are minimally absorbed and washed out of the intestinal lumen for application as antisecretory agents in cholera. METHODS: GlyH analogs (malonic hydrazides, MalH) were chemically conjugated to various lectins ("MalH-lectin") and purified. CFTR inhibition potency was measured by short-circuit current analysis, mechanism of action by patch-clamp, and antidiarrheal efficacy in closed-loop and suckling mouse models. RESULTS: By lectin conjugation, we improved CFTR inhibitory potency by approximately 100-fold (to 50 nmol/L) and retarded washout. High-affinity CFTR inhibition was abolished by MalH-lectin heat denaturation, protease digestion, or competition by mannose or unconjugated lectin. Patch-clamp analysis indicated CFTR inhibition by an external pore occlusion mechanism. Fluorescently labeled MalH-lectin remained membrane bound for >6 hours after washout, whereas washout occurred in a few minutes without the lectin. MalH-ConA and MalH-wheat (IC50 50 100 pmol) blocked cholera toxin-induced intestinal fluid secretion in closed intestinal loops in mice and greatly reduced mortality in a suckling mouse model of cholera. CONCLUSIONS: The high potency of MalH-lectin conjugates results from "anchoring" the CFTR-blocking MalH to cell surface carbohydrates by the lectin. The high-affinity, slow washout, and external site of action of the MalH-lectin conjugates support their further development as antisecretory drugs for enterotoxin-mediated secretory diarrheas. PMID- 17408660 TI - Long-term survival following endoscopic and surgical treatment of high-grade dysplasia in Barrett's esophagus. AB - BACKGROUND & AIMS: Photodynamic therapy (PDT) for high-grade dysplasia (HGD) in Barrett's esophagus is a Food and Drug Administration-approved alternative to esophagectomy. Critical information regarding overall survival of patients followed up long-term after these therapies is lacking. Our aim was to compare the long-term survival of patients treated with PDT with patients treated with esophagectomy. METHODS: We reviewed records of patients with HGD seen at our institution between 1994 and 2004. PDT was performed 48 hours following the intravenous administration of a photosensitizer using light at 630 nm. Esophagectomy was performed by either transhiatal or transthoracic approaches by experienced surgeons. We excluded all patients with evidence of cancer on biopsy specimens. Vital status and death date information was queried using an institutionally approved Internet research and location service. Statistical analysis was performed using Kaplan-Meier curves and Cox proportional hazards ratios. RESULTS: A total of 199 patients were identified. A total of 129 patients (65%) were treated with PDT and 70 (35%) with esophagectomy. Overall mortality in the PDT group was 9% (11/129) and in the surgery group was 8.5% (6/70) over a median follow-up period of 59 +/- 2.7 months for the PDT group and 61 +/- 5.8 months for the surgery group. Overall survival was similar between the 2 groups (Wilcoxon test = 0.0924; P = .76). Treatment modality was not a significant predictor of mortality on multivariate analysis. CONCLUSIONS: Overall mortality and long-term survival in patients with HGD treated with PDT appears to be comparable to that of patients treated with esophagectomy. PMID- 17408661 TI - Activation of Abl by Helicobacter pylori: a novel kinase for CagA and crucial mediator of host cell scattering. AB - BACKGROUND & AIMS: The pathogenesis of Helicobacter pylori (Hp)-associated diseases depends on a specialized type IV secretion system. This type IV secretion system injects the cytotoxin-associated gene A (CagA) effector protein into target cells where CagA becomes phosphorylated on tyrosine residues by Src. Src then is inactivated rapidly, suggesting the presence of another host tyrosine kinase to ensure constant CagA phosphorylation in sustained Hp infections. We aimed to identify this kinase. METHODS: By using the AGS gastric epithelial cell model, we performed a detailed functional characterization of Abl tyrosine kinase in signaling during Hp infections. RESULTS: We showed that Abl kinase is activated and a novel crucial mediator of Hp infections. First, Abl-specific inhibitors SKI-DV2-43 or STI571 (Gleevec, Novartis) and knockdown of c-Abl/Abl related gene Arg by small hairpin and interfering RNAs efficiently inhibit CagA phosphorylation and cell scattering. Second, during infection, Abl is activated rapidly by autophosphorylation at Y-412. Third, both Abl and Src phosphorylated Y 899, Y-918, and Y-972 of CagA. Fourth, we found that the Abl substrate CrkII is phosphorylated at Y-221 in vivo. Fifth, overexpression of kinase-dead Abl (K290M) blocked Hp-induced actin cytoskeletal rearrangements. We further showed that sustained activity of Abl is required to maintain CagA in a phosphorylated state. Moreover, phosphorylated CagA forms a physical complex with Abl and activated CrkII in vivo. CONCLUSIONS: We propose a model in which Hp has evolved a mechanism to use at least 2 tyrosine kinases, Abl and Src, for CagA phosphorylation and subsequent actin-cytoskeletal rearrangements leading to cell scattering and elongation. PMID- 17408662 TI - SCH 503034, a novel hepatitis C virus protease inhibitor, plus pegylated interferon alpha-2b for genotype 1 nonresponders. AB - BACKGROUND & AIMS: SCH 503034 is a novel and potent oral hepatitis C virus (HCV) protease inhibitor. In this phase Ib study, we assessed safety parameters and virologic response of combination of SCH 503034 plus pegylated (PEG) interferon (IFN) alpha-2b in patients with HCV genotype 1 infections who were previously nonresponders to PEG-IFN-alpha-2b +/- ribavirin therapy. METHODS: This was a multicenter, open-label, 2-dose level, 3-way crossover, randomized (to crossover sequence) study carried out in 3 medical centers in Europe. Adult patients received SCH 503034 200 mg (n = 14) or 400 mg (n = 12) 3 times daily orally and PEG-IFN-alpha-2b 1.5 microg/kg subcutaneously once each week. Patients received SCH 503034 as monotherapy for 1 week, PEG-IFN-alpha-2b as monotherapy for 2 weeks, and combination therapy for 2 weeks with washout periods between each treatment period. RESULTS: Combination therapy with SCH 503034 and PEG-IFN-alpha 2b was well tolerated, with no clinically significant changes in safety parameters. Mean maximum log(10) changes in HCV RNA were -2.45 +/- 0.22 and -2.88 +/- 0.22 for PEG-IFN-alpha-2b plus 200 mg and 400 mg SCH 503034, respectively, compared with -1.08 +/- 0.22 and -1.61 +/- 0.21 for SCH 503034 200 mg and 400 mg, respectively, and -1.08 +/- 0.22 and -1.26 +/- 0.20 for PEG-IFN-alpha-2b alone in the 200 mg and 400 mg SCH 503034 groups, respectively. CONCLUSIONS: SCH 503034 plus PEG-IFN-alpha-2b was well tolerated in patients with HCV genotype 1 nonresponders to PEG-IFN-alpha-2b +/- ribavirin. These preliminary results of antiviral activity of the combination suggest a potential new therapeutic option for this hard-to-treat, nonresponder patient population. PMID- 17408663 TI - Helminth infection enhances disease in a murine TH2 model of colitis. AB - BACKGROUND & AIMS: There is convincing evidence from animal and human studies that infection with parasitic helminths can alleviate the histopathology and symptoms of colitis. Here the ability of the rat tapeworm Hymenolepis diminuta to affect the course of oxazolone-induced colitis (a TH2 model) was assessed. METHODS: Mice were infected with H diminuta and 8 days later they received oxazolone (3 mg in 50% EtOH, intrarectal). On autopsy (3 or 7 days postoxazolone), disease severity was assessed by macroscopic clinical scores, histologic damage scores, myeloperoxidase and eosinophil peroxidase activity, and cytokine synthesis. RESULTS: As gauged by all markers of gut function, infection with H diminuta caused a significant exacerbation of oxazolone-induced colitis. Indeed, while mice receiving oxazolone only began to recover approximately 3-4 days posttreatment, the cotreated group continued to deteriorate. Helminth infection, independent of oxazolone administration, enhanced IL-4, IL-5, IL-10, and IL-13 production from in vitro stimulated immune cells and evoked increases in colonic eosinophil peroxidase of cotreated mice. Finally, while knockout of natural killer (NK) and NK-T cells by administration of a neutralizing NK1.1 antibody reduced the inflammation in oxazolone and oxazolone + H diminuta-treated animals, mice in the latter group still displayed significant colitis. CONCLUSIONS: We have shown that H diminuta infection is beneficial in other models of colitis. The current data is presented as a caveat to the position that parasitic helminths in general can be considered as a therapy for heterogeneous inflammatory disorders without careful analysis of the immunologic basis of the condition. PMID- 17408664 TI - Aberrant epigenetic modifications in hepatocarcinogenesis induced by hepatitis B virus X protein. AB - BACKGROUND & AIMS: The involvement of the hepatitis B virus X (HBx) protein in epigenetic modifications during hepatocarcinogenesis has not been previously characterized. The aim of the present study was to identify the involvement of HBx in regional hypermethylation and global hypomethylation during the formation of hepatocellular carcinoma (HCC). METHODS: Liver cell lines were transiently or stably transfected with an HBx-expressing vector. DNA methyltransferase (DNMT) promoter activity changes were examined by luciferase assay and chromatin immunoprecipitation. The methylation status of insulin-like growth factor binding protein-3 was examined by methyl-specific polymerase chain reaction and bisulfite sequencing. Global DNA methylation levels were examined using 5-methylcytosine dot blot and methylation-sensitive Southern blot analysis. HBx-mediated DNA methylation abnormalities were confirmed in patient HCC samples using methyl specific polymerase chain reaction and 5-methylcytosine dot blot analysis. RESULTS: HBx expression increased total DNMT activities by up-regulation of DNMT1, DNMT3A1, and DNMT3A2 and selectively promoted regional hypermethylation of specific tumor suppressor genes. HBx specifically repressed insulin-like growth factor-3 expression through de novo methylation via DNMT3A1 and DNMT3A2 and by inhibiting SP1 binding via recruiting methyl CpG binding protein 2 to the newly methylated SP1 binding element. HBx also induced global hypomethylation of satellite 2 repeat sequences by down-regulating DNMT3B. The prevalence of these specific methylation abnormalities by HBx was significantly correlated with HBx expression in HBV-infected HCC patients. CONCLUSIONS: Targeted deregulation of DNMTs by HBx promotes both specific regional hypermethylation and global hypomethylation. These epigenetic modulations by HBx may suggest a mechanism for epigenetic tumorigenesis during HBV-mediated hepatocarcinogenesis. PMID- 17408665 TI - Humoral immune response to tissue transglutaminase is related to epithelial cell proliferation in celiac disease. AB - BACKGROUND & AIMS: Tissue transglutaminase (tTG) autoantibodies are markers of celiac disease, and the enzyme is required for several crucial biological processes. The aim of this study was to determine whether these autoantibodies are involved in the pathogenesis of the mucosal lesion typical of celiac disease. METHODS: Using rhodamine-conjugated phalloidin staining, we evaluated whether tTG antibodies, both commercially available and cloned from patients with celiac disease, cause cytoskeletal changes in Caco-2, MCF7, and NIH 3T3 cells. We monitored cell levels of bromodeoxyuridine incorporation to determine whether tTG autoantibodies are able to induce NIH 3T3 fibroblasts and epithelial mucosal cells into S phase. RESULTS: Treatment with tTG antibodies caused a dose dependent increase of membrane ruffling in Caco-2, MCF7, and NIH 3T3 cells. It also dose-dependently induced G(0)-synchronized NIH 3T3 fibroblasts into S phase but did not affect the rate of apoptosis. Similarly, tTG antibodies induced S phase entry of epithelial cells in cultured intestinal biopsy specimens from patients with celiac disease. They did not affect biopsy specimens from patients without celiac disease. CONCLUSIONS: Our results suggest that tTG autoantibodies per se, by interacting with the extracellular membrane-bound transglutaminase, may play an important role in epithelial cell proliferation in celiac disease. PMID- 17408666 TI - Metabolism of cydiastatin 4 and analogues by enzymes associated with the midgut and haemolymph of Manduca sexta larvae. AB - The degradation of synthetic cydiastatin 4 (ARPYSFGL-amide) and cydiastatin 4 analogues cydiastatin 4alpha (PPPPPARPYSFGL-amide) and cydiastatin 4beta (PPPPPARPYSF[Acpc]L-amide) by enzymes associated with the midgut and/or haemolymph of the tobacco hawkmoth moth, Manduca sexta was investigated using reversed-phase high performance liquid chromatography (RP-HPLC) combined with matrix assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF MS). Cydiastatin 4 had an estimated half-life of c. 16.5min when incubated with midgut tissue in vitro and c. 2.5min with midgut lumen contents. Two degradation products were identified; cydiastatin(1-6), due to cleavage of the C-terminal di-peptide GL-amide, and cydiastatin(2-8), due to cleavage of the N-terminal A residue. Both cydiastatin 4alpha and cydiastatin 4beta had increased stability to gut and haemolymph enzymes, and full biological activity, but reduced potency compared to cydiastatin 4 when assayed on foregut peristalsis. The P-extended N-terminus of both analogues prevented hydrolysis by aminopeptidases and the replacement of the susceptible G residue with cyclopropylalanine ([Acpc]) counteracted carboxypeptidase activity. However, both analogues were susceptible to amidase-like activity giving an increase in one mass unit presumably due to the conversion of the C-terminal amide group to the free carboxylic acid. No metabolism of cydiastatin 4beta occurred when incubated with larval M. sexta haemolymph over a 90min period. PMID- 17408667 TI - Seasonal changes in cortisol and progesterone secretion in Common hamsters. AB - In this study, we investigated endocrine factors and behaviour in free-living Common hamsters (Cricetus cricetus) during reproductive and non-reproductive periods of the annual cycle. We applied a non-invasive method to gain information on seasonal changes in adrenocortical activity in male and female hamsters by analysing faecal glucocorticoid metabolite concentrations (FCM). In addition, plasma progesterone concentrations were monitored in females throughout the non hibernation season. The animals were live-trapped from spring emergence until the onset of hibernation in autumn. Reproductive status was determined at capture and blood and faecal samples were collected. During behavioural observations, agonistic and sexual interactions were recorded. FCM concentrations were significantly higher in males than in females during the reproductive period. In males, a pronounced increase in FCM during the reproductive period coincided with high frequencies of intrasexual aggression. In females, FCM levels remained relatively constant. Aggressive behaviour in females increased during the reproductive period, but was much less frequent than in males. Females, which successfully raised a second litter after a postpartum oestrus and concurrent lactation and gestation had lower FCM levels than individuals, which lost their second litter after parturition. As expected, plasma progesterone concentrations were low before and after the reproductive period. During gestation, levels peaked and remained elevated during lactation. The results of this field study provide insight in critical periods associated with reproduction in male and female Common hamsters. PMID- 17408668 TI - Timeliness of care for eclampsia and pre-eclampsia in Benin, Ecuador, and Jamaica. AB - OBJECTIVE: Cases meeting diagnostic criteria for severe pre-eclampsia or eclampsia were reviewed in three countries to determine timeliness and effectiveness of care. METHOD: Cases were retrospectively selected from 11 emergency obstetric care facilities and medical records reviewed by trained obstetricians. RESULT: Of 91 cases (Benin, 28; Ecuador, 25; Jamaica, 38), 74% were correctly treated with anticonvulsant and 77% with antihypertensive therapy. The median interval to treat eclampsia (anticonvulsant, 28 min; antihypertensive, 77 min) was shorter than for severe pre-eclampsia (anticonvulsant, 45 min; antihypertensive, 85 min). Two in three cases (65%) received anticonvulsant but only 41% received antihypertensive therapy within 60 min of diagnosis. While 74% of eclamptics had been delivered within 12 h, only 39% of severe pre-eclamptics were delivered within 24 h. CONCLUSION: Timeliness can be studied in developing countries. Its objective measurement is a first step towards improving this component of care. PMID- 17408669 TI - Risk factors for pelvic organ prolapse. AB - OBJECTIVES: To evaluate the risk factors for pelvic organ prolapse (POP) and to determine the relationship between these risk factors and stage or other components of POP. METHODS: 244 patients with primary POP and 314 women without POP were included. Age, parity, smoking, body mass index (BMI), menopause, and hormone replacement therapy (HRT) were investigated. RESULT: Independent risk factors for POP included age over 70, parity higher than 3, and menopause. Age, parity, menopause, and HRT were significantly associated with stage of POP. Genital hiatus (GH) and perineal body (PB) showed a significant positive and negative correlation with age and parity, respectively. Menopause and HRT were also associated with them. CONCLUSION: Age, parity and menopause are possible risk factors of POP and associated with the lengths of GH and PB in POP women. Further, these risk factors and HRT are significantly correlated with the severity of the disease. PMID- 17408670 TI - Birth outcomes of Asian-Indian-Americans. AB - OBJECTIVE: This study examines the maternal characteristics and birth outcomes of infants of U.S. resident Asian-Indian-American (AIA) mothers and compares those to infants of U.S. resident Whites and African-American (AA) mothers. METHODS: Single live births to U.S. resident mothers with race/ethnicity coded on birth certificate as AIA, non-Hispanic White, or non-Hispanic AA were drawn from NCHS 1995 to 2000 U.S. Linked Live Birth/Infant Death files. RESULTS: Compared to AAs or Whites, AIAs have the lowest percentage of births to teen or unmarried mothers and mothers with high parity for age or with low educational attainment. After taking these factors into account, AIA had the highest risk of LBW, small-for gestational age and term SGA births but a risk of infant death only slightly higher than Whites and far less than AAs. CONCLUSIONS: The birth outcomes of AIAs do not follow the paradigm that more impoverished minority populations should have greater proportions of low birth weight and preterm births and accordingly greater infant mortality rates. PMID- 17408671 TI - Nurses' requirements for information technology: a challenge for educators. PMID- 17408672 TI - The expression of caring within the perioperative dialogue: a hermeneutic study. AB - BACKGROUND AND AIM: This article, which is based on a study conducted in a perioperative context in Sweden, focuses on nurse anaesthetists' and theatre nurses' descriptions of caring and how caring becomes visible to patients in the perioperative dialogue. The perioperative dialogue is the pre-, intra- and postoperative meeting between the nurse and patient in connection with the latter's surgery. METHODS: Semi-structured interviews were conducted with 18 patients and 20 nurses, in which they described their experiences of perioperative dialogues. The interpretation process was based on Gadamer's philosophy of hermeneutics. FINDINGS: The findings indicate that, as a caring act, the vow involves: promising to allow the patient to be him/herself, promising to safeguard the welfare of the patient and promising to guide the patient through the operation as well as taking responsibility for that which I have promised. From the patient's perspective, the nurse was someone who: cares about me, takes me seriously, creates a calm atmosphere and to whom I can hand over responsibility. CONCLUSIONS: Four empirical assumptions were formulated in relation to the nurse's vow and how the nurse's caring became visible to the patients. PMID- 17408673 TI - A case for whole-parasite malaria vaccines. AB - Malaria causes morbidity in 300-500 million people each year and claims 2-3 millions lives annually, mostly children in sub-Saharan Africa. In 1983, the cloning of malaria antigens offered great promise for developing a viable subunit vaccine. However, an efficacious human vaccine is still not available. Immunological studies on how the host's immune system interacts with the parasite and studies on the pathogenic aspect of Plasmodium have found that several factors can impede protection by current vaccines. These findings suggest a novel approach needs to be considered. PMID- 17408674 TI - Can periods of static loading be used to enhance the resistance of the spine to cumulative compression? AB - Results of in vitro studies conducted on isolated bone specimens have indicated a higher tolerance to static load than exists when exposed to cyclic loading, when controlled for creep rate. If this difference in load tolerance exists, it may be exploited to extend the life of vertebral bone exposed to repetitive compression, and potentially alter the development of spinal injury. However, little work has been conducted on functional spinal units to determine if bone displays this characteristic within an intact joint. Additionally, static loading may result in load redistribution within the intervertebral disc forcing more of the compressive load towards the periphery of the endplate away from the nucleus. In order to examine these potential mechanisms, 218 osteoligamentous porcine functional spinal units were assigned to one of 15 loading scenarios. This involved one of three normalized peak load magnitudes (50%, 70% and 90% of estimated compressive tolerance) and one of five normalized static load applications (0%, 50%, 100%, 200% and 1000% of the total dynamic work duration). Load magnitude significantly altered the resistance to cumulative compression with decreased peak magnitudes corresponding to both increased cumulative load tolerance and increased height loss. Static load periods did not alter the resistance of the spinal unit to cumulative compression or impact the number of cycles tolerated to failure. The insertion of static load periods impacted the total survival time to failure, but only for the 1000% static load group, an exposure unlikely to occur for most in vivo exposures. The insertion of static load periods decreased the amount of height loss during testing which may play a protective role by allowing load redistribution within the vertebral bone and intervertebral disc. PMID- 17408675 TI - 3D finite element analyses of insertion of the Nucleus standard straight and the Contour electrode arrays into the human cochlea. AB - Previous experimental studies of insertion of the Nucleus standard straight and the Contour arrays into the scala tympani have reported that the electrode arrays cause damage to various cochlear structures. However, the level of insertion induced damage by these electrode arrays to cochlear structures (the spiral ligament, the basilar membrane and the osseous spiral lamina) has not been quantified. Although it has been suggested that rotation can overcome this resistance and prevent the basilar membrane from being pierced by the tip of the Nucleus standard straight array, there has not been any attempt to study the relationship between the rotation and the reduction of damage to the basilar membrane. In this study, 3D finite element analyses of insertions of the Nucleus standard straight array and the Contour array into the scala tympani have been undertaken. The perforation of the basilar membrane by the tip of the Nucleus standard straight array at the region of 11-14 mm from the round window appears to be compounded by the geometry of the spiral passage of the scala tympani. Anti clockwise rotations between 25 degrees and 90 degrees applied at the basal end of the electrode array (for the right cochlea) were shown to significantly reduce the contact stresses exerted by the tip on the basilar membrane which support the practice of applying small rotation partway through insertion of electrode array to minimize damage to the basilar membrane. Although the Contour array (with its stylet intact) is stiffer than the Nucleus standard straight array, a slight withdrawal of the stylet from the Contour array before insertion was found to significantly reduce damage by the electrode array to the spiral ligament and the basilar membrane. PMID- 17408676 TI - Model for spiral columns and stationary phase retention in synchronous coil planet centrifuges. AB - Stationary phase retention in a synchronous coil planet centrifuge or high-speed counter-current chromatography (CCC) relies on the interplay of hydrostatic (tangential and normal centrifugal) and hydrodynamic (Archimedean screw and mobile phase drag) forces. By offering a set of quantitative or semi-quantitative theoretical frameworks, this work has resolved fundamental questions such as "in the absence of mobile phase flow, how is the distribution of the two phases in a CCC column determined?" and "for Type-J CCC, do the helical and the spiral columns lead to similar performance?" PMID- 17408678 TI - Development of an affinity silica monolith containing alpha1-acid glycoprotein for chiral separations. AB - An affinity monolith based on silica and containing immobilized alpha(1)-acid glycoprotein (AGP) was developed and evaluated in terms of its binding, efficiency and selectivity in chiral separations. The results were compared with data obtained for the same protein when used as a chiral stationary phase with HPLC-grade silica particles or monoliths based on a copolymer of glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA). The surface coverage of AGP in the silica monolith was 18% higher than that obtained with silica particles and 61% higher than that measured for a GMA/EDMA monolith. The higher surface area of the silica monolith gave materials that contained 1.5- to 3.6 times more immobilized protein per unit volume when compared to silica particles or a GMA/EDMA monolith. The retention, efficiency and resolving power of the AGP silica monolith were evaluated by injecting two chiral analytes onto this column (i.e., R/S-warfarin and R/S-propranolol). In each case, the AGP silica monolith gave higher retention plus better resolution and efficiency than AGP columns containing silica particles or a GMA/EDMA monolith. The AGP silica monolith also gave lower back pressures and separation impedances than these other materials. It was concluded that silica monoliths can be valuable alternatives to silica particles or GMA/EDMA monoliths when used with AGP as a chiral stationary phase. PMID- 17408679 TI - High performance liquid chromatography/tandem mass spectrometry determination of biogenic amines in typical Piedmont cheeses. AB - The paper presents a new HPLC method, hyphenated with mass spectrometry detection, for the separation and determination of the biogenic amines that are most commonly present in cheese, namely cadaverine, histamine, spermidine, spermine, tyramine and tryptamine. The HPLC-MS/MS method is validated by comparison of the results with those obtained through a literature HPLC-UV determination, based on a pre-column dansyl chloride derivatisation step. The intercalibration is based on the statistical t-test for multiple samples that allows to compare simultaneously the results obtained with the two methods for more analytes and to decide, at a prefixed confidence level, if the two methods are inter-changeable. The new HPLC-MS/MS method, employed in selected reaction monitoring (SRM) mode, permits to achieve for standard solutions limit of detection (LOD) values ranging from 1.7 to 22.5 microg L(-1) and LOQ (limit of quantitation) values ranging from 5.6 to 68.2 microg L(-1). In order to apply the method in the analysis of cheeses, LOD and LOQ values have also been evaluated in "ricotta" cheese, in order to take as possible into account the matrix interference. In these conditions LODs range between 5.1 and 35.0 microg L(-1) and LOQs between 14.2 and 101.2 microg L(-1). The whole methodology, comprehensive of the homogenization-extraction process and HPLC-MS/MS analysis, has been applied in the analysis of three typical Piedmont (North-West Italy) cheeses, known as Toma Piemontese, Raschera and Castelmagno. PMID- 17408680 TI - Calibration, uncertainty, and recovery in the chromatographic sciences. AB - This paper reviews calibration-, uncertainty-, and recovery-related documents from 10 consensus-based organizations. The main points from each treatise are summarized. Also included is a critique of the various approaches, as well as recommendations for a statistically sound protocol that is more compatible with chromatographic data. PMID- 17408681 TI - Applications of counter-current chromatography in organic synthesis purification of heterocyclic derivatives of lapachol. AB - This work describes the application of counter-current chromatography (CCC) as a useful, fast and economic alternative for the isolation and purification of heterocyclic derivatives from lapachol and beta-lapachone, two naturally occurring compounds from Tabebuia species, and nor-beta-lapachone, a synthetic congener of lapachol. The discussed data comprise four examples of purification of synthetic reactions with different solvent systems - the mixture of the oxazole and the imidazole from beta-lapachone; the quinoxaline from nor-beta lapachone; and the purification of the N-oxides from the quinoxaline and the phenazine from nor-beta-lapachone from their respective not fully reacted substrates by means of aqueous reversed- and normal-phase elution modes and non aqueous solvent systems. Traditional purification of these reaction products by silica gel column chromatography demanded a large amount of solvent and time and, in some cases, serious degradation of the products occurred, leading to low yield of the reaction. High-speed counter-current chromatography (HSCCC) was used as an alternative to optimize the process and raise the yield of the reactions. PMID- 17408682 TI - Determination of beta-blockers and beta2-agonists in sewage by solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - A method using solid-phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the determination of 12 beta blockers and beta(2)-agonists in wastewater samples. Extraction of the drugs was effected by an Oasis MCX cartridge with a strong cation resin adsorbent. Matrix coextractives were removed from the SPE cartridge by methanol prior to the elution of the drugs with a mixture of dichloromethane, 2-propranol, and ammonium hydroxide. The extract was analyzed by LC-MS/MS with electrospray ionization operating in the positive mode. Recovery of the 12 compounds was in most cases better than 85% at the fortification levels of 500 and 50 ng/L, with standard deviations between 3 and 7%. Based on a concentration factor of 250, the method detection limits ranged from 6 to 11 ng/L for the target compounds. No degradation of these drugs in spiked sewage effluent samples was observed over a storage period of 7 days at 4 degrees C in the dark. Many beta-blockers were detected in the 14 wastewater samples collected from seven Canadian sewage treatment plants; those occurring at the highest concentrations were atenolol, acebutolol, sotalol, and metroprolol, with overall median concentrations of 1370, 339, 282, and 257 ng/L, respectively. However, there was only a small decrease in the median concentrations of these beta-blockers between the primary and final effluent, suggesting that these drugs are not easily removed by the sewage treatment processes. As a result of selective extraction and effective removal of coextractives, no matrix effect was observed for the samples during LC-MS/MS analyses. PMID- 17408683 TI - Modelling counter-current and dual counter-current chromatography using longitudinal mixing cell and eluting counter-current distribution models. AB - Two well known approaches are considered to analyze the processes of counter current and dual counter-current chromatography: the longitudinal mixing cell model and the Craig's counter-current distribution model. The cell model represents perfectly mixed, equally sized cells in series. The number of cells characterizes the rates of longitudinal mixing in the stationary and mobile phases. In the eluting counter-current distribution (CCD) model, the CCC process is considered as a continuous form of Craig's counter-current distribution. For a cascade of equilibrium stages theoretical elution profiles of the CCC process by using the CCD and cell model approaches have been compared. It is shown that in general, distribution functions of the CCD and cell models differ. It is established that the distribution of a solute between two solvent phases in the dual CCC process is determined by the extraction factor c, the total number of equilibrium stages n and the position of the sample inlet m by the equation Q(x)=(1-c(m))/(1-c(n+1)) with c=F(2)K(D)/F(1) (K(D), F(1), F(2) and Q(x) are the distribution constant, the phase flow-rates and the portion of solute eluted by the first phase, respectively). PMID- 17408684 TI - Prediction of solid-water-hydrocarbon contact angle. AB - The reliability of a recently developed solid-vapour and solid-liquid interfacial tension models has been investigated by applying them to predict liquid-vapour and liquid-liquid interfacial tension values. The impact of the geometrical molecular packing and the molecular orientations near the surface on the predicted values are discussed. The mutual solubility data are shown to be adequate for calculation of the interaction parameters in the solid-liquid model and a new equation, using this information, is developed for prediction of water hydrocarbon interfacial tension. The model has been applied to recent data on water-methane-n-decane and water-methane-cyclohexane-n-decane interfacial tensions at elevated temperature and pressure and its reliability demonstrated. It is shown that the solid-liquid interfacial tension model is solely adequate for predicting the contact angle by applying it to mercury-water-benzene and stearic acid-water-n-decane systems. PMID- 17408685 TI - The trouble with nouns and verbs in Greek fluent aphasia. AB - In the past verb retrieval problems were associated primarily with agrammatism and noun retrieval difficulties with fluent aphasia. With regards to fluent aphasia, so far in the literature, three distinct patterns of verb/noun dissociations have been described for individuals with fluent anomic aphasia in languages with different underlying forms; better verb retrieval, poorer verb retrieval and equal retrieval difficulties for verbs and nouns. Verbs and nouns in Greek are considered of similar morphological complexity thus it was predicted that anomic aphasic individuals would suffer from a non-dissociated impairment of verbs and nouns. Problems with verbs and/or nouns may arise at any stage in the process of lexical retrieval, i.e. lexical-semantic, lemma, lexeme or articulation. The aim of this research was to investigate verb and noun retrieval using a picture-naming task to explore any possible selective noun and/or verb comprehension or retrieval deficits in Greek individuals with anomic aphasia. The results revealed a significant verb/noun dichotomy with verbs significantly more difficult to retrieve than nouns. These findings lend support for the growing body of evidence showing a specific verb impairment in fluent anomic individuals as well as Broca's patients. Given the prevailing view, that anomic patients experience difficulty retrieving the morpho-phonological form of the target word, the results show that specific information of the grammatical category is also important during word form retrieval. LEARNER OUTCOMES: The reader will become familiar with (i) studies investigating grammatical word class breakdown in individuals with aphasia who speak different languages, (ii) the application of the serial model to word production breakdown in aphasia and (iii) the characteristics of verbs and nouns in Greek. It will be concluded that successful verb retrieval for fluent aphasic individuals who speak Greek is dependant on the retrieval of the morpho-phonological information of the target verb. PMID- 17408686 TI - Tracking the acquisition of orthographic skills in developing readers: masked priming effects. AB - A masked priming procedure was used to explore developmental changes in the tuning of lexical word recognition processes. Lexical tuning was assessed by examining the degree of masked form priming and used two different types of prime target lexical similarity: one letter different (e.g., rlay-->PLAY) and transposed letters (e.g., lpay-->PLAY). The performance of skilled adult readers was compared with that of developing readers in Grade 3. The same children were then tested again two years later, when they were in Grade 5. The skilled adult readers showed no form priming, indicating that their recognition mechanisms for these items had become finely tuned. In contrast, the Grade 3 readers showed substantial form priming effects for both measures of lexical similarity. When retested in Grade 5, the developing readers no longer showed significant one letter different priming, but transposed letter priming remained. In general, these results provide evidence for a transition from more broadly tuned to more finely tuned lexical recognition mechanisms and are interpreted in the context of models of word recognition. PMID- 17408687 TI - Generation of mutant leukaemia inhibitory factor (LIF)-IgG heavy chain fusion proteins as bivalent antagonists of LIF. AB - Two leukaemia inhibitory factor (LIF) mutants, designated MH35-BD and LIF05, have been shown to have a capacity to inhibit the biological activities of not only human LIF (hLIF) but also other interleukin-6 (IL-6) subfamily cytokines such as human oncostatin M (hOSM). These cytokines share the same receptor complex in which the glycoprotein 130 (gp130) subunit is a common constituent. However, at low concentrations and in their monomeric forms, such molecules have a relatively short plasma half-life due to rapid clearance from the kidneys. Here, to prolong their serum half-lives, we have used a multi-step polymerase chain reaction (PCR) to fuse each of the LIF05 and MH35-BD cDNA fragments to a sequence encoding the Fc portion, and the hinge region, of the human immunoglobulin G (hIgG) heavy chain. The linking was achieved through an oligomer encoding a thrombin-sensitive peptide linker thus generating MH35-BD:Fc and LIF05:Fc, respectively. Both Fc fusion constructs were expressed in insect cell Sf21 and the proteins were purified by two successive affinity chromatography steps using nickel nitrilotriacetic acid (Ni-NTA) agarose and protein A beads. The Ba/F3 cell-based proliferation assay was used to confirm that the proteins were biologically active. In addition, preliminary pharmacokinetics indicates that the Fc fusion constructs have a longer serum half-life compared to their non-fusion counterparts. PMID- 17408688 TI - Stress and high heart rate provoke ventricular tachycardia in mice expressing triadin. AB - Reduced function of the cardiac ryanodine receptor or calsequestrin causes catecholaminergic ventricular tachycardia (VT). These proteins regulate sarcoplasmic Ca(2+) release in close conjunction with two accessory proteins, triadin and junctin. Based on data from cardiomyocytes, we hypothesized that enhanced triadin expression could cause VT. We assessed arrhythmias and electrophysiological changes in vivo and in the beating heart in mice expressing junctin, triadin, or both proteins (TRDxJCN), and measured calcium transients in isolated ventricular cardiomyocytes. TRDxJCN mice were studied to compensate the down-regulation of junctin expression in triadin-expressing mice. Exercise or stress provoked repetitive VT in freely roaming TRDxJCN mice whenever heart rate increased above approximately 600 bpm (p<0.05 vs. the three other genotypes). TRDxJCN mice expressed total triadin 2.9-fold (p<0.05) and total junctin not different to wildtype (p=ns). Left ventricular systolic function was not different between lineages. beta-adrenoreceptor stimulation (orciprenaline 1.7 microM) provoked early-coupled ventricular ectopy and repetitive VT in isolated, Langendorff-perfused TRDxJCN hearts (p<0.05). Under conditions associated with VT (high pacing rate, catecholamine stimulation), action potential duration was shorter in TRDxJCN with VT than in the other genotypes and shorter than in TRDxJCN hearts without VT (p<0.05). Ca(2+) transient duration was prolonged in Indo1-loaded TRDxJCN cardiomyocytes under VT-provoking conditions. Action potential prolongation by mexiletine (2 microM or 4 microM) or clarithromycine (150 microM) suppressed VT. Expression of triadin provokes stress- and tachycardia-related ventricular arrhythmias in mice. An imbalance between prolonged intracellular calcium release and shortening of the ventricular action potential may contribute to genesis of arrhythmias in this model. PMID- 17408689 TI - Defining how ubiquitin receptors hHR23a and S5a bind polyubiquitin. AB - Ubiquitin receptors connect substrate ubiquitylation to proteasomal degradation. HHR23a binds proteasome subunit 5a (S5a) through a surface that also binds ubiquitin. We report that UIM2 of S5a binds preferentially to hHR23a over polyubiquitin, and we provide a model for the ternary complex that we expect represents one of the mechanisms used by the proteasome to capture ubiquitylated substrates. Furthermore, we demonstrate that hHR23a is surprisingly adept at sequestering the ubiquitin moieties of a polyubiquitin chain, and provide evidence that it and the ubiquitylated substrate are committed to each other after binding. PMID- 17408690 TI - Rapid folding and unfolding of Apaf-1 CARD. AB - Caspase recruitment domains (CARDs) are members of the death domain superfamily and contain six antiparallel helices in an alpha-helical Greek key topology. We have examined the equilibrium and kinetic folding of the CARD of Apaf-1 (apoptotic protease activating factor 1), which consists of 97 amino acid residues, at pH 6 and pH 8. The results showed that an apparent two state equilibrium mechanism is not adequate to describe the folding of Apaf-1 CARD at either pH, suggesting the presence of intermediates in equilibrium unfolding. Interestingly, the results showed that the secondary structure is less stable than the tertiary structure, based on the transition mid-points for unfolding. Single mixing and sequential mixing stopped-flow studies showed that Apaf-1 CARD folds and unfolds rapidly and suggest a folding mechanism that contains parallel channels with two unfolded conformations folding to the native conformation. Kinetic simulations show that a slow folding phase is described by a third conformation in the unfolded ensemble that interconverts with one or both unfolded species. Overall, the native ensemble is formed rapidly upon refolding. This is in contrast to other CARDs in which folding appears to be dominated by formation of kinetic traps. PMID- 17408691 TI - Complete sequence determination combined with analysis of transposition/site specific recombination events to explain genetic organization of IncP-7 TOL plasmid pWW53 and related mobile genetic elements. AB - Recent studies have indicated that the evolutionarily common catabolic gene clusters are loaded on structurally diverse toluene-catabolic (TOL) plasmids and their residing transposons. To elucidate the mechanisms supporting the diversification of catabolic plasmids and transposons, we determined here the complete 107,929 bp sequence of pWW53, a TOL plasmid from Pseudomonas putida MT53. pWW53 was found to belong to the IncP-7 incompatibility group that play important roles in the catabolism of several xenobiotics. pWW53 carried two distinct transposase-resolvase gene clusters (tnpAR modules), five short terminal inverted repeats (IRs), and three site-specific resolution (res) sites that are all typical of class II transposons. This organization of pWW53 suggested the four possible transposable regions, Tn4657 to Tn4660. The largest 86 kb region (Tn4657) spanned the three other regions, and Tn4657 and Tn4660 (62 kb) covered all of the 36 xyl genes for toluene catabolism. Our subsequent transposition experiments clarified that the three transposons, Tn4657 to Tn4659, indeed exhibit their transposability, and that pWW53 also generated another 37 kb toluene-catabolic transposon, Tn4656, which carried the two separated and inversely oriented segments of pWW53: the tnpRA-IR module of Tn4658 and a part of xyl gene clusters on Tn4657. The Tn4658 transposase was able to mediate the transposition of Tn4658, Tn4657, and Tn4656, while the Tn4659 transposase catalyzed only the transposition of Tn4659. Tn4656 was formed by the Tn4658 resolvase-mediated site-specific inversion between the two inversely oriented res sites on pWW53. These findings and comparison with other catabolic plasmids clearly indicate multiple copies of transposition-related genes and sites on one plasmid and their recombination activities contribute greatly to the diversification of plasmid structures as well as wide dissemination of the evolutionary common gene clusters in various plasmids. PMID- 17408692 TI - No association found between the promoter variants of ADRA1A and schizophrenia in the Chinese population. AB - Schizophrenia is a chronic psychiatry disorder with a strong genetic component. A recent association study of alpha(1A)-adrenoceptor gene (ADRA1A) involving an isolated Spanish population, focusing on the promoter region of the ADRA1A, genotyped eight single SNPs at the promoter region of ADRA1A and found that two SNPs, -563G/A and -9625G/A, were associated with schizophrenia and schizoaffective disorders. We were interested in the two positive sites reported and selected five variants among the promoter region of ADRA1A, namely -563G/A, 9625G/A, -2760C/A, -4155G/C and a new substitution we detected between -508bp and -530bp upstream of the translation initiation site. Our sample consisted of 480 schizophrenia and 480 control subjects. All recruits were Han Chinese in Shanghai origin. However, neither individual SNP nor any haplotype was associated with schizophrenia in our study. These results suggest that the variants among the promoter of ADRA1A gene are unlikely to play a major role in the susceptibility to schizophrenia in the Chinese population. PMID- 17408693 TI - Association of schizophrenia with DTNBP1 but not with DAO, DAOA, NRG1 and RGS4 nor their genetic interaction. AB - Recent reports indicate that DAO, DAOA, DTNBP1, NRG1 and RGS4 are some of the most-replicated genes implicated in susceptibility to schizophrenia. Also, the functions of these genes could converge in a common pathway of glutamate metabolism. The aim of this study was to evaluate if each of these genes, or their interaction, was associated with schizophrenia. A case-control study was conducted in 589 Spanish patients having a diagnosis of schizophrenia, and compared with 617 equivalent control subjects. Several single nucleotide polymorphisms (SNPs) in each gene were determined in all individuals. SNP and haplotype frequencies were compared between cases and controls. The interaction between different SNPs at the same, or at different gene, loci was analyzed by the multifactor dimensionality reduction (MDR) method. We found a new schizophrenia risk and protective haplotypes in intron VII of DTNBP1; one of the most important candidate genes for this disorder, to-date. However, no association was found between DAO, DAOA, NRG1 and RGS4 and schizophrenia. The hypothesis that gene-gene interaction in these five genes could increase the risk for the disorder was not confirmed in the present study. In summary, these results may provide further support for an association between the dysbindin gene (DTNBP1) and schizophrenia, but not between the disease and DAO, DAOA, NRG1 and RGS4 or with the interaction of these genes. In the light of recent data, these results need to be interpreted with caution and future analyses with dense genetic maps are awaited. PMID- 17408694 TI - Earlier onset and shorter survival of amyotrophic lateral sclerosis in Jewish patients of North African origin. A clue to modifying genetic factors? AB - Amyotrophic lateral sclerosis (ALS) is a multifactorial disease, with many genetic and environmental factors contributing to its outcome. The population of Israel is comprised of immigrants from all over the world as well as by Arabs. People with different ethnic backgrounds who live in the same environment provide a unique opportunity to analyze genetic and environmental influences on ALS. We performed a retrospective analysis of 374 sporadic ALS patients whose origin was European in 211, North African in 53, Oriental in 43, Balkan in 19, Arab in 9, and Yemenite in 7, comparing their age at disease onset, gender, disease form at onset, survival, smoking habits, cognitive dysfunction and apolipoprotein E genotype. Patients of North African origin were significantly younger and had a shorter duration of disease relative to their age compared to other ethnic groups, adjusted for age. The difference between the patient groups might be related to a genetic burden in North African patients and warrants further investigation. PMID- 17408695 TI - Stroke as the first manifestation of concealed cancer. AB - BACKGROUND AND PURPOSE: Trousseau's syndrome (TS) is defined as a malignancy related thromboembolism occurring in patients with an underlying or undiagnosed malignancy. Stroke seldom occurs as the first manifestation of a cancer. We investigated the clinical and radiological features of patients with TS. METHODS: We reviewed the clinical, pathologic, and radiological records of consecutive stroke patients, whose cancers were diagnosed at stroke presentation. Cancer related stroke was defined if no definite cause was confirmed and malignancy was detected within 6 months of first stroke onset without cancer-related treatment. All patients underwent brain diffusion-weighted MRI (DWI), MR angiography, and echocardiography. The sizes, numbers, and locations of all hyperintense lesions in the DWI were noted. RESULTS: Ten patients were finally analyzed. Histologically, cancers were often proven to adenocarcinomas (50%, 5/10) of advanced stage. Six of 10 patients tested (60%) had elevated D-dimer. Seven of the 10 patients (70%) showed bihemispheric anterior and posterior involvement. DWI features showed numerous small and medium or large lesions in multiple territories in 9 patients (90%). CONCLUSIONS: The authors emphasize that when presented with multiple bihemispheric infarctions on DWI and an unknown etiology, the neurologist must consider the existence of a concealed cancer. PMID- 17408696 TI - A new technique for the mapping of oxygen tension on the brain surface. AB - Most measurements of oxygen tension (PO(2)) in the brain have been performed using oxygen microelectrodes. However, the insertion of microelectrodes into the brain per se causes cortical injury and hence could lead to erroneous PO(2) measurements. The recently developed "quenching lifetime method" requires the injection of fluorescent chemicals into the blood circulation. To address this issue, we tested the feasibility of our O(2)-sensitive fluorescent membrane technique in the rat brain, and visualized the spatial distribution of PO(2) on the brain surface as epifluorescent microscopic patterns. An O(2)-quenching fluorescence dye, tris (1,10-phenanthroline) Ru(2+), was immobilized in a highly gas-permeable, thin silicone-rubber film formed on a microscope coverslip. Unlike the original method, which was intended for transparent rat mesenteric tissue, any change in the redox state in the brain tissue will influence the optical measurement of PO(2). Thus, in the present study, the O(2)-sensing membrane was further coated with a thin opaque silicone-rubber to minimize this type of influence. This new method enabled us to visualize the PO(2) gradient on the rat brain without causing cortical injuries. In an ischemia/reperfusion model using Pulsinelli's four-vessel occlusion rats, the changes in the PO(2) were highly heterogeneous during the ischemic period and this heterogeneity, both temporal and spatial, was higher in the off-arteriolar area than in the peri-arteriolar area. PMID- 17408697 TI - Boolean dynamics of Kauffman models with a scale-free network. AB - We studied the Boolean dynamics of the "quenched" Kauffman models with a directed scale-free network, comparing with that of the original directed random Kauffman networks and that of the directed exponential-fluctuation networks. We have numerically investigated the distributions of the state cycle lengths and its changes as the network size N and the average degree k of nodes increase. In the relatively small network (N approximately 150), the median, the mean value and the standard deviation grow exponentially with N in the directed scale-free and the directed exponential-fluctuation networks with k=2, where the function forms of the distributions are given as an almost exponential. We have found that for the relatively large N approximately 10(3) the growth of the median of the distribution over the attractor lengths asymptotically changes from algebraic type to exponential one as the average degree k goes to k=2. The result supports the existence of the transition at k(c)=2 derived in the annealed model. PMID- 17408698 TI - Effect of temperature on viral infection and its control: a mathematical approach. AB - The interrelationship of the host-virus-environment is of critical importance in determining the amount of virus production per insect and its consequences in the overall infection process in insect community. Lower temperature inhibits the virus replication and thus infection does not persist into the system. On the other hand, virus production per insect increases with temperature increase within certain limit, and this increase of virus production brings some unforeseen consequences in the infection dynamics in pest control. We deal the problem by applying non-vulnerability concept on the mathematical model of viral infection that linked up with the temperature-viral developmental model. Using Lyapunov-like function, we determine a range of temperature and show that the system would become endemic and remain in stable situation if temperature could be maintained in this prescribed range, whereas that range depends on other parameters of the system. PMID- 17408699 TI - Regional differences in the neuroprotective effect of minocycline in a mouse model of global forebrain ischemia. AB - We investigated the effect of minocycline on neuronal damage in the hippocampus and striatum in a mouse model of transient global forebrain ischemia. Male C57BL/6 mice were anesthetized with halothane and subjected to bilateral occlusion of the common carotid artery (BCCAO) for 30 min. Minocycline (90 mg/kg, i.p., qd) or saline was injected immediately after BCCAO and daily for the next two days (45 mg/kg, i.p., bid). In order to reduce the variability in ischemic neuronal damage, we applied selection criteria based on regional cerebral blood flow (rCBF), evaluated using laser Doppler flowmetry, and the plasticity of the posterior communicating artery (PcomA), evaluated using India ink solution. In animals with rCBF that was less than 15% of the baseline value and with a smaller PcomA, of diameter less than one-third that of the basilar artery, we consistently observed neuronal damage in the striatum and hippocampal subfields, including medial CA1, CA2, and CA4. When the effect of minocycline was assessed with cresyl violet staining, neuronal damage in the medial part of the CA1 subfield and the striatum was found to be significantly attenuated, although minocycline did not protect against neuronal damage in the remaining hippocampal subfields. Immunohistochemistry for NeuN, adenosine A1 receptor, and SCIP/Oct-6 confirmed the region-specific effect of minocycline in the hippocampus. In summary, our results suggest that minocycline protects neurons against global forebrain ischemia in a subregion-specific manner. PMID- 17408700 TI - The magic glue hyaluronan and its eraser hyaluronidase: a biological overview. AB - Hyaluronan (HA) is a multifunctional high molecular weight polysaccharide found throughout the animal kingdom, especially in the extracellular matrix (ECM) of soft connective tissues. HA is thought to participate in many biological processes, and its level is markedly elevated during embryogenesis, cell migration, wound healing, malignant transformation, and tissue turnover. The enzymes that degrade HA, hyaluronidases (HAases) are expressed both in prokaryotes and eukaryotes. These enzymes are known to be involved in physiological and pathological processes ranging from fertilization to aging. Hyaluronidase-mediated degradation of HA increases the permeability of connective tissues and decreases the viscosity of body fluids and is also involved in bacterial pathogenesis, the spread of toxins and venoms, acrosomal reaction/ovum fertilization, and cancer progression. Furthermore, these enzymes may promote direct contact between pathogens and the host cell surfaces. Depolymerization of HA also adversely affects the role of ECM and impairs its activity as a reservoir of growth factors, cytokines and various enzymes involved in signal transduction. Inhibition of HA degradation therefore may be crucial in reducing disease progression and spread of venom/toxins and bacterial pathogens. Hyaluronidase inhibitors are potent, ubiquitous regulating agents that are involved in maintaining the balance between the anabolism and catabolism of HA. Hyaluronidase inhibitors could also serve as contraceptives and anti-tumor agents and possibly have antibacterial and anti-venom/toxin activities. Additionally, these molecules can be used as pharmacological tools to study the physiological and pathophysiological role of HA and hyaluronidases. PMID- 17408701 TI - Triiodothyronine (T3)-mediated toxicity and induction of apoptosis in insulin producing INS-1 cells. AB - Thyroid hormones reduce glucose tolerance in humans and animals. This effect is related to a decrease of glucose-induced insulin secretion following a reduction of pancreatic beta cell mass due to beta cell loss. The aim of this study was to analyze in vitro the mechanisms underlying the effects of triiodothyronine (T(3)) on the cell viability and cell cycle caused by changes of cell death or proliferation rate of insulin-producing INS-1 cells. 72-h Exposure of INS-1 cells to increasing T(3) concentrations up to 500 microM resulted in a significant viability reduction. This T(3) toxicity was caused by an increased apoptotic cell death rate, which was accompanied by a decreased proliferation rate. Inhibitory effects of T(3) on glucose-induced insulin secretion were already seen after 24 h of incubation, indicating that the deleterious effects of T(3) were time dependent, changing from specific cellular dysfunctions to a severe and extended disturbance of the cellular survival program. Only T(3) concentrations higher than 250 microM were able to decrease cell viability and proliferation rate, to increase the rate of apoptosis and to reduce glucose-induced insulin secretion. These micromolar T(3) concentrations were significantly higher than the concentration range of T(3) receptor binding, indicating that other non-receptor mediated mechanisms beyond the receptor level must be responsible for the observed toxic effects of T(3) in vitro. PMID- 17408702 TI - Air pollution by carcinogenic PAHs and plasma levels of p53 and p21(WAF1) proteins. AB - We analyzed the effect of exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) in ambient air on the plasma levels of p53 and p21(WAF1) proteins among city policemen, bus drivers and controls in three European cities: Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). p53 and p21(WAF1) proteins are key regulators of the cell cycle and are accepted as universal markers of genotoxic stress and DNA damage. In total 204 exposed subjects (100 smokers, 104 nonsmokers) and 152 controls (54 smokers, 98 nonsmokers) were analyzed. Personal exposure to c-PAHs was evaluated using personal samplers during the working shift. The levels of p53 and p21(WAF1) proteins were assessed by ELISA assay. There were no differences between the levels of either protein between exposed and controls, or smokers and nonsmokers, in any city. However, we observed significant differences in p53 plasma levels in all subjects regardless of the exposure status between the individual cities (median values: 5, 31, 234pg/ml, p<0.001, for Prague, Kosice and Sofia, respectively). The levels correspond to the differences in exposure levels to c PAHs and benzo[a]pyrene (B[a]P) in the individual cities. A multiple linear regression analysis confirmed that c-PAHs exposure is a variable significantly affecting levels of both proteins in all locations. When all subjects were divided into the group exposed to below-median levels of c-PAHs and the group exposed to above-median levels of c-PAHs we found significantly higher p53, as well as p21(WAF1) levels in the above-median exposure group (p53, 167pg/ml versus 25pg/ml, p<0.001; p21(WAF1), 2690pg/ml versus 2600pg/ml, p<0.05). Among all subjects p53 plasma levels were positively correlated with p21(WAF1) levels, exposure to B[a]P, c-PAHs and levels of total DNA adducts; for p21(WAF1) levels we observed the positive correlation with cotinine, c-PAHs exposure, total and B[a]P-like DNA adduct levels. In conclusion our results suggest that p53 and p21(WAF1) proteins plasma levels may be useful biomarkers of c-PAHs environmental exposure. PMID- 17408703 TI - GSTM1, GSTT1 and CYP1A1 detoxification gene polymorphisms and susceptibility to smoking-related coronary artery disease: a case-only study. AB - Cigarette smoking is a powerful risk factor for coronary artery disease (CAD), leading to the formation of DNA alterations within blood vessels and heart. However, the degree of smoking-related atherosclerosis varies from individual to individual. Genetic polymorphisms of relevant xenobiotic metabolising enzymes may determine the susceptibility of an individual response to environmental toxicants. The purpose of this study was to test the hypothesis that the inheritance of polymorphic genes encoding cytochrome P450 1A1 (CYP1A1 MspI) and glutathione S-transferases (GSTM1(null) and GSTT1(null)) may be causally associated with the presence and severity of smoking-induced CAD. In a case-only design, 222 (179 male, 57.8+/-10.3 years) consecutive smoker patients who had undergone elective and diagnostic coronary angiography were recruited. We found a group (n=169) of smoker patients with significant CAD, defined as>50% reduction in diameter of at least one major vessel, and a group without obstructive CAD (n=53). No significant differences were observed in CYP1A1 genotypes frequencies between CAD and non-CAD smokers (p=0.1). Homozygous deletion of GSTM1 had a frequency of 58.6% among patients with CAD and 45.3% among those without CAD (p=0.08). The frequency of the GSTT1(null) genotype was 43.8% among the patients with CAD and 24.5% among CAD-free subjects (p=0.01). After adjustment for traditional risk factors, the presence of combined GSTM1(null)GSTT1(null) genotypes was significantly associated with an increased risk of CAD (OR=3.9; 95% CI: 1.3-11.4, p=0.01). Moreover, smokers with combined GSTM1(null)GSTT1(null) genotypes had significantly higher number of stenosed vessels than those with the positive genotype (2.3+/-0.9 versus 1.7+/-0.8, p=0.03). Our findings showed that smokers carrying GST deleted genotypes have an increased susceptibility to the smoking related coronary artery disease. Exploring gene-smoking effect provides an excellent model in order to understand gene-environment toxicants interaction and its implications to cardiovascular disease. PMID- 17408704 TI - Amygdala activation and facial expressions: explicit emotion discrimination versus implicit emotion processing. AB - Emotion recognition is essential for social interaction and communication and is a capacity in which the amygdala plays a central role. So far, neuroimaging results have been inconsistent as to whether the amygdala is more active during explicit or incidental facial emotion processing. In consideration of its functionality in fast automatic evaluation of stimuli and involvement in higher order conscious processing, we hypothesize a similar response to the emotional faces presented regardless of attentional focus. Using high field functional magnetic resonance imaging (fMRI) specifically optimized for ventral brain regions we show strong and robust amygdala activation for explicit and implicit processing of emotional facial expressions in 29 healthy subjects. Bilateral amygdala activation was, however, significantly greater when subjects were asked to recognize the emotion (explicit condition) than when required to discern the age (implicit condition). A significant correlation between amygdala activation and emotion recognition, but not age discrimination performance, emphasizes the amygdala's enhanced role during conscious emotion processing. PMID- 17408705 TI - Urine from stressed rats increases immobility in receptor rats forced to swim: role of 2-heptanone. AB - The present study was aimed to determine whether the urine from donor rats, which were physically stressed (UD-PS) by unavoidable electric footshocks, produces despair in receptor partner rats (RP) in the long-term. For each trial, an RP rat was placed during 10 min once per day for 21 days in a small non-movement restricting cage impregnated with the urine collected from a UD-PS rat. Control rats, free of stimulation, maintained their locomotion and immobility scores at basal values throughout the 21-day test. After 21 days of stressing experience [F(2,90)=15.22, P<0.0001] locomotion significantly increased in RP rats (r=0.938, P<0.01), whereas in the UD-PS group locomotion decreased (r=-0.606, P<0.05). The RP and UD-PS groups displayed the longest time of immobility [F(2,90)=8.83, P<0.001] in the forced-swim test (RP, r=0.886, P<0.05; UD-PS, r=0.962, P<0.001) compared with the control group (r=-0.307, NS). We conclude that the RP became similarly despaired as the UD-PS group through the action of 2-heptanone, a ketonic compound identified in UD-PS urine by HS-GC/MS techniques. This ketone was found to be increased [F(2,15)=3.50, P<0.05] from the 1st day of unavoidable electric footshocks, and to induce despair, an effect reverted [F(2,21)=16.5, P<0.0001] by imipramine (5.0 mg/kg) in another group of rats. PMID- 17408706 TI - Isolation and structure determination of triterpenes from Iris tectorum. AB - Four iridal-type triterpenoids, two of which were new compounds, have been isolated from rhizomes of Iris tectorum Maxim. Their structures were determined by 1D and 2D NMR spectroscopy and ESI-MS spectrometry. The compounds were identified as the iritectols A and B, and the known iridobelamal A and isoiridogermanal. The presence of epoxide and tetrahydrofuran functions are not common in previously isolated iridal-type triterpenoids. PMID- 17408707 TI - Triterpenoid saponins with N-acetyl sugar from the bark of Albizia procera. AB - Three (1,2,4) and one known (3) triterpenoid saponins were isolated from the bark of Albizia procera. The saponins were characterized as 3-O-[beta-D-xylopyranosyl (1-->2)-alpha-L-arabinopyranosyl-(1-->6)-2-acetamido-2-deoxy-beta-D glucopyranosyl] echinocystic acid (1), 3-O-[alpha-L-arabinopyranosyl-(1-->2)-beta D-fucopyranosyl-(1-->6)-2-acetamido-2-deoxy-beta-D-glucopyranosyl] echinocystic acid (2) and 3-O-[beta-D-xylopyranosyl-(1-->2)-alpha-L-arabinopyranosyl-(1-->6)-2 acetamido-2-deoxy-beta-D-glucopyranosyl] acacic acid lactone (4). Their structures were elucidated by 1D and 2D NMR experiments, FABMS as well as chemical means. Saponins 1 and 3 exhibited cytotoxicity against HEPG2 cell line with IC50 9.13 microg/ml and 10 microg/ml, respectively. PMID- 17408708 TI - The N-terminal region of the starch-branching enzyme from Phaseolus vulgaris L. is essential for optimal catalysis and structural stability. AB - Starch-branching enzymes (SBEs) play a pivotal role in determining the fine structure of starch by catalyzing the syntheses of alpha-1,6-branch points. They are the members of the alpha-amylase family and have four conserved regions in a central (beta/alpha)8 barrel, including the catalytic sites. Although the role of the catalytic barrel domain of an SBE is known, that of its N- and C-terminal regions remain unclear. We have previously shown that the C-terminal regions of the two SBE isozymes (designated as PvSBE1 and PvSBE2) from kidney bean (Phaseolus vulgaris L.) have different roles in branching enzyme activity. To understand the contribution of the N-terminal region to catalysis, six chimeric enzymes were constructed between PvSBE1 and PvSBE2. Only one enzyme (1Na/2Nb)-II, in which a portion of the N-terminal region of PvSBE2 was substituted by the corresponding region of PvSBE1, retained 6% of the PvSBE2 activity. The N terminal truncated form (DeltaN46-PvSBE2), lacking 46 N-terminal residues of PvSBE2, lost enzyme activity and stability to proteolysis. To investigate the possible function of this region, three residues (Asp-15, His-24, and Arg-28) among these 46 residues were subjected to site-directed mutagenesis. The purified mutant enzymes showed nearly the same K(m) values as PvSBE2 but had lower V(max) values and heat stabilities than PvSBE2. These results suggest that the N terminal region of the kidney bean SBE is essential for maximum enzyme activity and thermostability. PMID- 17408709 TI - Antiplasmodial hirsutinolides from Vernonia staehelinoides and their utilization towards a simplified pharmacophore. AB - The dichloromethane extract of the leaves of Vernonia staehelinoides Harv. (Asteraceae) showed in vitro activity (IC(50) approximately 3 microg/ml) against the chloroquine-sensitive (D10) and the chloroquine-resistant (K1) strains of Plasmodium falciparum. Through conventional chromatographic techniques and bioassay-guided fractionation two structurally-related hirsutinolides displaying in vitro antiplasmodial activity (IC(50) approximately 0.2 microg/ml against D10) were isolated and identified by spectroscopic data. Compounds 1, 8 alpha-(2 methylacryloyloxy)-3-oxo-1-desoxy-1,2-dehydrohirsutinolide-13-O-acetate, and 2, 8 alpha-(5'-acetoxysenecioyloxy)-3-oxo-1-desoxy-1,2-dehydrohirsutinolide-13-O acetate were found to be cytotoxic to mammalian Chinese Hamster Ovarian (CHO) cells at similar concentrations but proved to be attractive scaffolds for structure-activity relationship studies. Two main privileged substructures, a 2(5H)-furanone unit and a dihydrofuran-4-one unit, were identified as potential pharmacophores which may be responsible for the observed biological activity. Mucochloric and mucobromic acids were selected as appropriate 2(5H)-furanone substructures and these were shown to have comparable activity against the D10 and superior activity against the K1 strains relative to the hirsutinolide natural product. Mucochloric and mucobromic acids also show selective cytotoxicity to the malaria parasites compared to mammalian (CHO) cells in vitro. The antiplasmodial data obtained in respect of these two acids suggests that the 2(5H)-furanone substructure is a key pharmacophore in the observed antiplasmodial activity. PMID- 17408710 TI - Effect of medetomidine infusion on the anaesthetic requirements of desflurane in dogs. AB - The objective of this paper was to evaluate the effect of constant rate infusion of medetomidine on the anaesthetic requirements of desflurane in dogs. For this, six healthy dogs were studied. Measurements for baseline were taken in the awake, unsedated dogs, then each dog received intravenously (i.v.) three anaesthetic protocols: M (no medetomidine infusion), M0.5 (infusion of medetomidine at 0.5 microg/kg/h, i.v.) or M1 (infusion of medetomidine at 1 microg/kg/h, i.v.). All dogs were sedated with medetomidine (2 microg/kg, i.v.) and measurements repeated in 10 min. Induction of anaesthesia was delivered with propofol (3 mg/kg, i.v.) and maintained with desflurane for 90 min to achieve a defined surgical plane of anaesthesia in all cases. After tracheal intubation infusion of medetomidine was initiated and maintained until the end of anaesthesia. Cardiovascular, respiratory, arterial pH (pHa) and arterial blood gas tensions (PaO(2), PaCO(2)) variables were measured during the procedure. End tidal desflurane concentration (EtDES) was recorded throughout anaesthesia. Time to extubation, time to sternal recumbency and time to standing were also noted. Heart rate and respiratory rate were significantly decreased during sedation in all protocols compared to baseline values. Mean heart rate, mean arterial pressure, systolic arterial pressure, diastolic arterial pressure, respiratory rate, tidal volume, arterial oxygen saturation, end-tidal CO(2), pHa, PaO(2), and PaCO(2) during anaesthesia were similar for all protocols. EtDES for M (8.6 +/- 0.8%) was statistically higher than for M0.5 (7.6 +/- 0.5%) and M1 (7.3 +/- 0.7%) protocols. Infusion of medetomidine reduces desflurane concentration required to maintain anaesthesia in dogs. PMID- 17408711 TI - Comparison of the effects of firocoxib, carprofen and vedaprofen in a sodium urate crystal induced synovitis model of arthritis in dogs. AB - A randomized, placebo-controlled, four-period cross-over laboratory study involving eight dogs was conducted to confirm the effective analgesic dose of firocoxib, a selective COX-2 inhibitor, in a synovitis model of arthritis. Firocoxib was compared to vedaprofen and carprofen, and the effect, defined as a change in weight bearing measured via peak ground reaction, was evaluated at treatment dose levels. A lameness score on a five point scale was also assigned to the affected limb. Peak vertical ground reaction force was considered to be the most relevant measurement in this study. The firocoxib treatment group performed significantly better than placebo at the 3 h post-treatment time point and significantly better than placebo and carprofen at the 7 h post-treatment time point. Improvement in lameness score was also significantly better in the dogs treated with firocoxib than placebo and carprofen at both the 3 and 7 h post treatment time points. PMID- 17408712 TI - Molecular cloning of serine proteases from elapid snake venoms. AB - Serine proteases are widely distributed in viperid snake venoms, but rare in elapid snake venoms. Previously, we have identified a fibrinogenolytic enzyme termed OhS1 from the venom of Ophiophagus hannah. The results indicated that OhS1 might be a serine protease, but there was no structural evidence previously. In the present study, the primary structure of OhS1 was determined by protein sequencing, in combination with RT-PCR and 5'-RACE methods. OhS1 precursor is composed of an 18-amino acid signal peptide, a 6-amino acid putative activation peptide and 236-amino acid mature protein. OhS1 homologues from Naja atra and Bungarus multicinctus were also cloned and reported. These elapid venom serine proteases exhibited approximately 60% sequence identity with serine proteases from the snake venoms of the Viperidae and Colubridae family. Phylogenetic analysis indicated that snake venom serine protease might have a common ancestor. PMID- 17408713 TI - In vitro assembly of a prohead-like structure of the Rhodobacter capsulatus gene transfer agent. AB - The gene transfer agent (GTA) is a phage-like particle capable of exchanging double-stranded DNA fragments between cells of the photosynthetic bacterium Rhodobacter capsulatus. Here we show that the major capsid protein of GTA, expressed in E. coli, can be assembled into prohead-like structures in the presence of calcium ions in vitro. Transmission electron microscopy (TEM) of uranyl acetate staining material and thin sections of glutaraldehyde-fixed material demonstrates that these associates have spherical structures with diameters in the range of 27-35 nm. The analysis of scanning TEM images revealed particles of mass approximately 4.3 MDa, representing 101+/-11 copies of the monomeric subunit. The establishment of this simple and rapid method to form prohead-like particles permits the GTA system to be used for genome manipulation within the photosynthetic bacterium, for specific targeted drug delivery, and for the construction of biologically based distributed autonomous sensors for environmental monitoring. PMID- 17408714 TI - Pneumo- and neurotropism of avian origin Italian highly pathogenic avian influenza H7N1 isolates in experimentally infected mice. AB - An experimental infection of mice was performed in order to investigate the potential for interspecies transmission in mammals of Italian HPAI viruses of the H7N1 subtype. Three avian origin isolates were selected, two strains obtained from ostrich (one of which contained a PB2-627 Lysine residue) and one from a chicken. Following intranasal infection of mice, clinical signs and mortality were recorded in the experimental groups challenged with the two ostrich isolates, while only weight loss was observed in those receiving the chicken strain. Viruses were recovered to a varying extent from respiratory and nervous tissues of infected animals. These results suggest that HPAI viruses, other than H5N1 and H7N7, may have zoonotic implications, and support the consensus that AI infections in poultry are to be eradicated rather than contained. PMID- 17408715 TI - Serological detection systems for identification of cows shedding bovine foamy virus via milk. AB - The biology of foamy viruses, their mode of transmission and disease potential in their natural host and after interspecies transmission are largely unknown. To gain insights into the prevalence of bovine foamy virus (BFV) and its zoonotic potential, enzyme-linked immunosorbent assays (ELISAs) were established to determine antibody responses against Gag, Env, and the non-structural protein Bet in bovine serum and milk. In Polish cattle, strong Gag reactivity was most frequent (41.5%) and strongly associated with Bet antibodies, Env antibodies were less frequent. German cattle showed a low overall BFV antibody prevalence of 6.8%. Besides clearly BFV-positive animals, a substantial number of weakly reacting cattle were identified. BFV-specific antibodies were also detectable in milk. BFV was isolated from PBLs and milk cells of BFV-positive cattle but not from antibody-negative or weakly reacting animals. The implications of these findings for the potential interspecies transmission of BFV to humans will be discussed. PMID- 17408716 TI - Optimizing electrode positions and analysis strategies for multifocal VEP recordings by ROC analysis. AB - The multifocal visual evoked potential (mfVEP) is an important tool to test visual pathway function. The aim of this study was to optimize electrode positions in mfVEP recordings. For analysis we applied a receiver operating characteristic (ROC), a method that inherently corrects for multiple testing. We found that a combination of two perpendicular derivations-both straddling the inion-was the most effective recording setup. Adding more than two derivations did not significantly increase the sensitivity. Thus optimal mfVEP detection can be achieved with a fairly simple recording setup which may facilitate mfVEP recordings in basic research and clinical routine. PMID- 17408717 TI - Evidence for the existence of colour mechanisms producing unique hues as derived from a colour illusion based on spatio-chromatic interactions. AB - When its spatial frequency is high enough, a grid of grey horizontal strips presented on a coloured background may change its neutral colour. It was found that some background colours induce a strong illusion and some no illusion at all. The effect of the background colour on the illusion was studied for the spatial frequencies of 0.5, 2.5, 4, and 8 c/deg. Thirty chromaticities (evenly distributed across the colour gamut triangle) of the backgrounds in the equiluminant plane, and 24 in the ML plane (where S-contrast was zero), were tested. Five matches were made for each frequency and each background chromaticity. Viewing was binocular. For the low (0.5 c/deg) frequency strips, the backgrounds were found to induce the colour, if any, approximately complimentary to that of the background (i.e., chromatic simultaneous contrast). For the high (8 c/deg) frequency, most backgrounds induced only illusory colours close to unique hues (yellow, blue, and green), with a few backgrounds inducing a mixture of green with blue. Then, the method of adjustment was used to determine the unique hues for the same three observers. A remarkable similarity was found between unique hues and illusory loci, suggesting that the illusion is due to a difference in the spatial resolution of the post-receptor channels producing the unique hues. PMID- 17408718 TI - Attentional blink deficits observed in dyslexia depend on task demands. AB - The attentional blink (AB) refers to a deficit in the ability to identify a second target following a first target when both appear randomly within a rapid sequence of distractor items. The AB of five adults with dyslexia (ADys) was compared with that of a group of normal adult readers. Two tasks were completed which differed in the conceptual category of the target items (a red digit or letter) relative to the distractor items (all black digits). In the digit condition, all ADys cases showed a longer AB compared to the control group. In the letter condition, all participants showed improvement in accuracy compared to the digit condition, but three ADys cases continued to have a longer AB. The results suggest that (a) AB performance depends on task requirements, and (b) the attentional system is compromised in dyslexia. However, examination of individual case performance suggests that prolonged attentional dwell time is not a core deficit in dyslexia. The results also illustrate the limitations of group comparisons in small sample studies. PMID- 17408719 TI - Elucidation and short-term forecasting of microcystin concentrations in Lake Suwa (Japan) by means of artificial neural networks and evolutionary algorithms. AB - Non-supervised artificial neural networks (ANN) and hybrid evolutionary algorithms (EA) were applied to analyse and model 12 years of limnological time series data of the shallow hypertrophic Lake Suwa in Japan. The results have improved understanding of relationships between changing microcystin concentrations, Microcystis species abundances and annual rainfall intensity. The data analysis by non-supervised ANN revealed that total Microcystis abundance and extra-cellular microcystin concentrations in typical dry years are much higher than those in typical wet years. It also showed that high microcystin concentrations in dry years coincided with the dominance of the toxic Microcystis viridis whilst in typical wet years non-toxic Microcystis ichthyoblabe were dominant. Hybrid EA were used to discover rule sets to explain and forecast the occurrence of high microcystin concentrations in relation to water quality and climate conditions. The results facilitated early warning by 3-days-ahead forecasting of microcystin concentrations based on limnological and meteorological input data, achieving an r(2)=0.74 for testing. PMID- 17408720 TI - Biostability analysis for drinking water distribution systems. AB - The ability to limit regrowth in drinking water is referred to as biological stability and depends on the concentration of disinfectant residual and on the concentration of substrate required for the growth of microorganisms. The biostability curve, based on this fundamental concept of biological stability, is a graphical approach to study the two competing effects that determine bacterial regrowth in a distribution system: inactivation due to the presence of a disinfectant, and growth due to the presence of a substrate. Biostability curves are a practical, system specific approach for addressing the problem of bacterial regrowth in distribution systems. This paper presents a standardized algorithm for generating biostability curves and this will enable water utilities to incorporate this approach for their site-specific needs. Using data from pilot scale studies, it was found that this algorithm was applicable to control regrowth of HPC in chlorinated systems where AOC is the growth limiting substrate, and growth of AOB in chloraminated systems, where ammonia is the growth limiting substrate. PMID- 17408721 TI - Calculation of serum "total lipid" concentrations for the adjustment of persistent organohalogen toxicant measurements in human samples. AB - Persistent organohalogen toxicants such as 2,3,7,8-tetrachlorodibenzo-p-dioxin or polychlorinated biphenyls measured in human serum are often expressed on a lipid weight basis, most commonly by dividing the toxicants' concentration by the weight of total lipids in the sample. Therefore, the manner in which this lipid adjustment is calculated may influence the final reported result. Gravimetric total lipid assays have been used, but they are time-consuming and sometimes may be ill-defined. Consequently, alternative methods using enzymatic assays have been developed based on summing the individual lipid species measured. Recent reports, however, have suggested that significantly different total lipid results may be obtained when using alternative formulae in a summation approach. In this report, we summarize the results obtained from lipid measurements of nearly 900 samples made as part of a study of a group of older American men (mean age 62 years), and we compare our total lipid estimates obtained by using both our standard and "short" formula (the latter based on total cholesterol and triglycerides only) with results obtained using the recently proposed alternative formulae. Our findings indicate that both our long and short formulae provide similar estimates of serum total lipid concentrations, and that differences observed in lipid estimates when using the newer alternative summation methods may reflect differences in how the term "total lipid" is defined, especially with regard to the need to include the contribution of the weight of the cholesterol ester fatty acids in the calculation. PMID- 17408722 TI - Extended geometric method: a simple approach to derive adsorption rate constants of Langmuir-Freundlich kinetics. AB - A new and simple equation has been presented here for calculation of adsorption and desorption rate constants of Langmuir-Freundlich kinetic equation. The derivation of new equation is on the basis of extension and correction to the geometric method which has been presented by Kuan et al. [Kuan, W.-H., Lo, S.-L., Chang, C.M., Wang, M.K., 2000. A geometric approach to determine adsorption and desorption kinetic constants. Chemosphere 41, 1741-1747] for the kinetics of adsorption/desorption in aqueous solutions. The correction is to consider that the concentration of solute is not constant and changes as adsorption proceeds. The extension is that we applied Langmuir-Freundlich kinetic model instead of Langmuir kinetic model to consider the heterogeneity and therefore it is more applicable to the real systems. For solving Langmuir-Freundlich kinetic model, some geometric methods and also Taylor expansion were used and finally a simple and novel equation was derived (Eq. (20)) for calculation of adsorption rate constant. This new method was named "extended geometric method". The input data of the obtained equation can be simply derived from initial data of adsorption kinetics. Finally the adsorption of methyl orange onto granular activated carbon was carried out at dynamic and equilibrium conditions and the capabilities of extended geometric method were examined by the experimental data. PMID- 17408723 TI - Anaerobic biodegradation of 4-alkylphenols in a paddy soil microcosm supplemented with nitrate. AB - Anaerobic degradation of phenol, p-cresol, 4-n-propylphenol (n-PP), 4-i propylphenol (i-PP), 4-n-butylphenol (n-BP) and 4-sec-butylphenol (sec-BP) was observed in a paddy soil supplemented with nitrate. We detected the metabolites 4'-hydroxypropiophenone (HPP) from n-PP, 4-i-propenylphenol from i-PP, and 4-(1 butenyl)phenol and 4'-hydroxybutyrophenone (HBP) from n-BP. Compared with the original soils, Betaproteobacteria became predominant in the microcosm during the degradation of phenol and p-cresol whereas no remarkable change was observed in the community degrading propylphenols and butylphenols. The microcosm, however, did not degrade 4-t-butylphenol (t-BP), 4-t-octylphenol (t-OP) and 4-n octylphenol (n-OP). Paddy soil supplemented with sulfate or iron (III) as electron acceptors did not degrade phenol and 4-alkylphenols with the exception of the degradation of p-cresol in sulfate-reducing conditions. It was demonstrated for the first time that anaerobic microbial degradation of alkylphenols, in a paddy soil supplemented with nitrate as an electron acceptor, occurred via oxidation of the alpha carbon in the alkyl chain. PMID- 17408724 TI - PCBs and OCPs in human milk and selected foods from Luqiao and Pingqiao in Zhejiang, China. AB - This study was conducted to measure the levels of 23 PCB congeners and 6 organochlorine pesticides (OCPs) in human milk and three food types collected from Luqiao and Pingqiao in Zhejiang Province, China. An effort was also made to explore the potential health risk for the mothers and breast-fed infants living in these two localities. Luqiao was selected as the sampling site because it is the largest place for the disassembly of obsolete transformers and electrical waste in China. Pingqiao, located 100 km NW of Luqiao, is not known to be a place for any electronic or electrical waste and hence was chosen as the control site. Both localities are important agricultural places in the province. The organochlorines were measured in the samples using the GC-muECD technique. Micro EROD bioassay method was also used as a complement of the chemical analysis to estimate the TEQ levels of dioxin-like PCBs in human milk. The data showed that the human milk, rice, hen egg, and fish samples from Luqiao were more heavily contaminated with PCBs than those from Pingqiao, suggesting that the mothers and their breast-fed infants in Luqiao tended to receive greater exposure to PCBs than those living in Pingqiao. The OCP levels in the two localities were found comparable, suggesting that the major source of contamination with these pesticides was from their agricultural uses. Significant correlation (R2 = 0.87, P<0.001) of PCB TEQs was found between the bioassay and chemical analysis method, suggesting that micro-EROD is an effective method for comprehensive determination of TEQ levels in human milk. Comparison with literature data showed that the PCB levels in milk samples from Luqiao were significantly higher than those from localities in other Chinese provinces and comparable to those in developed or industrialized countries. PMID- 17408725 TI - Fibrinogen Novy Jicin and Praha II: cases of hereditary Aalpha 16 Arg-->Cys and Aalpha 16 Arg-->His dysfibrinogenemia. AB - INTRODUCTION: Various dysfibrinogenemias have been described worldwide. This paper describes two new cases of dysfibrinogenemia identified in the Czech Republic. MATERIALS AND METHODS: The proposita of fibrinogen Novy Jicin, a 12 year-old girl, presented with hemorrhagic complications, low Clauss fibrinogen level (0.3 g/l) and prolonged both thrombin (70.8 s) and reptilase (>180 s) time. Her mother and sister both presented with normal coagulation tests, normal fibrinogen level and reported no history of bleeding. The carriers of the fibrinogen Praha II were a 31-year-old man and his 11-year-old daughter. They both presented with low fibrinogen Clauss level (0.88 g/l) and prolonged thrombin and reptilase time. To identify the genetic mutation responsible for these dysfibrinogens, genomic DNA extracted from the blood was analyzed. The presence of the mutant chains in the circulation was determined by MALDI-TOF mass spectroscopy. Scanning electron micrographs of the patients' fibrin clots were obtained. RESULTS: The kinetics of fibrinopeptide release and fibrin polymerization were impaired for both fibrinogen Novy Jicin and Praha II. DNA sequencing showed heterogeneous fibrinogen Aalpha R16C mutation in the fibrinogen Novy Jicin case and heterogeneous fibrinogen Aalpha R16H in the fibrinogen Praha II case. The mutant chains were found to be expressed to the circulation by MALDI TOF mass spectroscopy. Scanning electron micrographs of the patient's fibrin clot were found to be abnormal. CONCLUSIONS: The case of dysfibrinogenemia Aalpha R16C fibrinogen Novy Jicin and the case of dysfibrinogenemia Aalpha R16H were found by routine coagulation testing and were genetically identified. PMID- 17408726 TI - Venous thromboembolism in ovarian cancer. AB - OBJECTIVE: To determine the incidence, time-course, and risk factors associated with the development of thromboembolism (VTE) in a population-based study of women with ovarian cancer. METHODS: Using the California Cancer Registry, cases diagnosed with ovarian cancer for a 6-year period were identified. These cases were linked with the California Patient Discharge Data Set to determine the incidence of VTE. Proportional hazards modeling was performed to analyze the strength of specified risk factors to predict development of VTE or death within 2 years. RESULTS: Among 13,031 cases with ovarian cancer, 5.2% were diagnosed with a VTE event within 24 months after diagnosis. The cumulative incidence varied from 1.4% among women with local stage disease to 6.7% among women with advanced disease. The person-time incidence rate of VTE decreased over time, with the highest rate noted during the first 3 months. In a multivariate model, significant risk factors for VTE included advancing age, increasing number of chronic comorbid conditions, more advanced cancer stage, invasive histology, and absence of any major surgery. For all stages of cancer, development of VTE within 2 years was a significant risk factor for decreased survival, and the magnitude of the risk was greatest among the cases diagnosed with localized disease (HR 4.7, 95% CI: 2.3-9.5). CONCLUSIONS: VTE occurred in a significant proportion of ovarian cancer patients and adversely impacted survival, particularly among cases with local or regional stage cancer. PMID- 17408727 TI - The role of cytoreductive surgery for colon cancer metastatic to the ovary. AB - OBJECTIVE: We sought to further elucidate the survival impact of cytoreductive surgery among patients with colon cancer metastatic to the ovary. METHODS: All women diagnosed with primary colon cancer metastatic to the ovary at a single institution from 1980 to 2005 were retrospectively identified. Survival analyses and comparisons were performed using Kaplan-Meier plots and the log rank test. RESULTS: A total of 39 patients with 40 cases of colon cancer metastatic to the ovary were identified. Patients with metastatic disease confined to the ovaries (n=11) had a median overall survival (OS) time of 61 months (range 15-120) compared to 17 months (range 0.5-73) for those with more extensive metastases (n=24) (p=0.0428). Patients undergoing optimal cytoreduction (residual < or =1 cm) had a median progression-free survival (PFS) of 11 months (range 0.5-120, n=26) compared to 2.5 months (range 0.5-12, n=9) for those receiving suboptimal cytoreduction (p=0.0001). Optimal cytoreduction was also associated with a significantly longer median OS (35 months, range 0.5-120) compared to suboptimal cytoreduction (median OS=7 months, range=0.5-17) (p<0.0001). The peri-operative mortality rate was 5%. Significant morbidity occurred in 10% of the cases. All major complications occurred in women with diffuse disease who underwent extensive cytoreductive surgery. CONCLUSIONS: The observation that optimal cytoreduction was associated with prolonged PFS and OS in both patients with localized ovarian and widespread metastases of colon cancer suggests a role for surgical management of metastatic colon cancer in women. PMID- 17408728 TI - Sparing of saphenous vein during inguinal lymphadenectomy for vulval malignancies. AB - OBJECTIVE: This work was set out to investigate the effect of saphenous vein preservation during inguinal lymphadenectomy for patients with vulval malignancies. METHODS: 64 patients with vulval malignancies were allocated into two groups depending on their clinical stages, with one of them (31 patients included) being subjected to sparing of saphenous vein and the other to saphenous vein ligated surgery while treated with inguinal lymphadenectomy. The operative time, blood loss, 5-year survival rate, short- and long-term postoperative complications, 5-year survival rate and groin recurrence were selected as the monitored parameters, through which the above two groups were compared with each other using t test, chi2 and life table analysis. RESULTS: (1) The median operative time for bilateral inguinal lymphadenectomy was 155 min (130-170 min) in the sparing group, compared to 140 min (120-170 min) in the excision group (P>0.05). The median intraoperative blood loss was 295 mL (100-450 mL) in the sparing group, and 270 mL (150-390 mL) in the excision group (P>0.05). (2) Short term lower extremity lymphedema occurred with 27 patients (43.5%) in the sparing group and 44 patients (66.7%) in the excision group (P<0.01). Still, short-term lower extremity phlebitis was observed with 7 patients (11.3%) in the sparing group while 17 developed phlebitis (25.8%) in the excision group (P<0.05). However, there was no statistical difference in postoperative fever, acute cellulites, seroma, or lymphocyst formation. (3) Long-term complication occurrence rate decreased by about 50% in patients subjected to saphenous vein sparing surgery compared with those to ligated surgery, while there was no remarkable difference between two groups in the occurrence rates of phlebitis and deep venous thrombosis (P>0.05). (4) The overall 5-year survival rate was 67.3%, with 66.7% and 68.0% for the excision group and the sparing group, respectively (P>0.05). CONCLUSION: The application of saphenous vein preservation technique during inguinal lymphadenectomy for patients with vulval malignancies could significantly decrease the occurrence rate of postoperative complications without compromising outcomes and should be widely put into clinical practice. PMID- 17408729 TI - Individual differences in novelty- and cocaine-induced locomotor activity as predictors of food-reinforced operant behavior in two outbred rat strains. AB - A goal of the current study was to determine if individual differences in cocaine induced locomotion, which has been shown in outbred Sprague-Dawley (SD) rats to be correlated with differential function of dopamine transporters, were also evident in Long-Evans (LE) rats. Another objective was to determine if differences in locomotion following exposure to novelty or cocaine predicted food reinforced behavior. Between-strain comparisons of open-field activity revealed similar effects of 10 mg/kg cocaine, although increases in rearing were prominent in LE rats. Both strains exhibited robust individual differences in cocaine induced locomotion, with nearly identical ambulatory behavior observed in low and high cocaine responders (LCRs and HCRs, respectively) from the two strains. In a cued-discrimination operant task, LE rats learned the contingency in fewer sessions, whereas SD rats obtained more food pellets at fixed ratio 10 and maintained higher progressive ratio (PR) breakpoints. HCRs from both strains also tended to maintain higher PR breakpoints; low and high responders to novelty (LR and HR, respectively) had no consistent differences in food-reinforced behavior. Overall, these studies suggest that wide individual differences in cocaine induced behavior are common to SD and LE strains and certain differences in food reinforced behavior are associated with HCRs compared to LCRs. PMID- 17408730 TI - The dopamine D3 receptor antagonist NGB 2904 increases spontaneous and amphetamine-stimulated locomotion. AB - The dopamine D3 receptor is believed to play an important role in regulation of rodent locomotor behavior, and has been proposed as a therapeutic target for substance abuse, psychotic disorders, and Parkinson's disease. One model of dopamine D3 receptor function, based on studies utilizing D3 receptor knockout mice and D3 receptor-preferring agonists, proposes that D3 receptor stimulation is inhibitory to psychostimulant-induced locomotion, in opposition to the effects of concurrent dopamine D1 and D2 receptor stimulation. Recent progress in medicinal chemistry has led to the development of highly-selective dopamine D3 receptor antagonists. In order to extend our understanding of D3 dopamine receptor's behavioral functions, we determined the effects of the highly selective dopamine D3 receptor antagonist NGB 2904 on amphetamine-stimulated and spontaneous locomotion in wild-type and dopamine D3 receptor knockout mice. NGB 2904 (26.0 microg/kg s.c.) enhanced amphetamine-stimulated locomotion in wild type mice, but had no measurable effect in dopamine D3 receptor knockout mice. Of a range of doses (0.026 microg-1.0 mg/kg) given acutely or once daily for seven days, the highest dose of NGB 2904 (1.0 mg/kg) stimulated spontaneous locomotion in wild-type mice, but was without measurable effect in dopamine D3 receptor knockout mice. These behavioral effects of NGB 2904 contrast with those described for other highly D3 receptor-selective antagonists, which have not previously demonstrated an effect on spontaneous locomotor activity. In combination, these data add to the behavioral profile of this novel D3 receptor ligand and provide further support for a role for dopamine D3 receptor inhibitory function in the modulation of rodent locomotion. PMID- 17408732 TI - Expression of the GM-CSF receptor in ovine spermatozoa: GM-CSF effect on sperm viability and motility of sperm subpopulations after the freezing-thawing process. AB - The granulocyte-macrophage colony stimulating factor (GM-CSF) is a pleiotropic cytokine capable of stimulating proliferation, maturation and function of haematopoietic cells. Receptors for this cytokine are composed of two subunits, alpha and beta, and are expressed in myeloid progenitors and mature mononuclear phagocytes, monocytes, eosinophils and neutrophils, as well as in other non haematopoietic cells. We have previously demonstrated that bull spermatozoa express functional GM-CSF receptors that signal for increased glucose and vitamin C uptake and enhance several parameters of sperm motility in the presence of glucose or fructose substrates. In this study, we have analyzed the expression of GM-CSF receptors in ovine spermatozoa and studied the effect of GM-CSF on sperm viability and motility after the freezing-thawing process. Immunolocalization and immunoblotting analyses demonstrated that ovine spermatozoa (Xisqueta race) expressed GM-CSF receptors. In addition, GM-CSF partially counteracted the impairing action of freezing/thawing on the percentage of total motility, as well as on the specific motility patterns of each of the separate, motile sperm subpopulations of ram ejaculates subjected to this protocol. These results suggest that GM-CSF can play a role in the resistance of ram spermatozoa to environmental thermal stress. PMID- 17408733 TI - Omissions and semantic errors in aphasic naming: is there a link? AB - Within a discrete two-stage model of lexicalization, semantic errors and errors of omission are assumed to be independent events. In contrast, cascading and interactive models allow for an influence of word form on lexical selection and thus for an inherent relationship in accounting for both error types. A group of 17 aphasic patients was assessed with a naming test controlling for semantic competition of the target items. Semantic errors were more frequent for targets with many competitors than for targets with few competitors while omissions were more frequent when few competitors were available. However, the overall sums of errors in both item groups were comparable. These results imply a common source of both error types and thus speak against a strictly serial model of naming. PMID- 17408734 TI - Elution of monomers from two conventional dental composite materials. AB - OBJECTIVE: The aim of this study was to determine the elution of monomers of two conventional resin composite materials after different polymerization and storage times. METHODS: Two resin composites (a hybrid and a flowable) were used. Four groups (n=10, diameter: 4.5mm, thickness: 2mm) of each material were fabricated, one for each polymerization time of: 0s, 20s, 40s, and 80s. The samples were stored in 1ml of 75% ethanol at room temperature, and the storage medium was renewed after 24h, 7 days, and 28 days. From the storage medium that was removed, samples were prepared and evaluated, with LC-MS/MS. RESULTS: Bisphenol A and UDMA were not detected in the samples. Regardless of the polymerization time, the material or the storage time, a higher amount of BisGMA was eluted compared to TEGDMA. The amount of monomer that was released from the polymerized samples of the hybrid resin composite (Tetric Ceram) was significantly higher (p<0.0001) compared to the flowable (Tetric Flow). No significant difference was found between samples polymerized for 20s compared to 40s concerning the elution of monomers. Only a polymerization time of 80s resulted in a decreased release of monomers. The release of TEGDMA decreased after 28 days; however, the elution of BisGMA remained at high levels. SIGNIFICANCE: The release of monomers remains at a high level for a long time (7-28 days) after polymerization. The 40s that are usually used for the polymerization of resin composites seems insufficient in order to prevent a high release of monomers. PMID- 17408735 TI - Effect of copper on the scope for growth of clams (Tapes philippinarum) from a farming area in the Northern Adriatic Sea. AB - Copper is currently the most common biocide in antifouling paints. Levels of this metal were measured in the water, particulate matter and sediments from a shellfish farming area in the Sacca di Goro (Northern Adriatic Sea) over one year. With respect to the 1980s, copper environmental level increased twofold. The release of copper from shellfish farmers' boats was also estimated to be > 250 kg Cu y(-1). Clams Tapes philippinarum were collected in the same area and seasonally exposed to a sublethal (10 microg Cu l(-1)) concentration of copper. Physiological traits were significantly affected by copper exposure (scope for growth declined as a result of reduced clearance rate, increased oxygen consumption and a generally lower absorption efficiency). The results of this study are cause for concern for shellfish farming activities at least in Northern Adriatic, where shellfish farming is a monoculture of T. philippinarum. A strict interpretation of the precautionary principle might suggest that more rigorous regulatory action to control copper inputs in the field would be justified. PMID- 17408736 TI - Guidance of glial cell migration and axonal growth on electrospun nanofibers of poly-epsilon-caprolactone and a collagen/poly-epsilon-caprolactone blend. AB - Our long-term goal is to develop an artificial implant as a conduit for axonal regeneration after peripheral nerve injury. In this study, biodegradable, aligned poly-epsilon-caprolactone (PCL) and collagen/PCL (C/PCL) nanofibers designed as guidance structures were produced by electrospinning and tested in cell culture assays. We compared fibers of 100% PCL with fibers consisting of a 25:75% C/PCL blend. To test their biocompatibility, assays of cell adhesion, survival, migration, effects on cell morphology, axonal growth and axonal guidance were performed. Both types of eletrospun fibers supported oriented neurite outgrowth and glial migration from dorsal root ganglia (DRG) explants. Schwann cell migration, neurite orientation, and process formation of Schwann cells, fibroblasts and olfactory ensheathing cells were improved on C/PCL fibers, when compared to pure PCL fibers. While the velocity of neurite elongation from DRG explants was higher on PCL fibers, analysis of isolated sensory neurons showed significantly better axonal guidance by the C/PCL material. The data demonstrate that electrospun fibers composed of a collagen and PCL blend represent a suitable substrate for supporting cell proliferation, process outgrowth and migration and as such would be a good material for artificial nerve implants. PMID- 17408737 TI - Chitosan microparticles encapsulating PEDF plasmid demonstrate efficacy in an orthotopic metastatic model of osteosarcoma. AB - The major stumbling block for most therapies against deep-seated disease, including tumours, is inefficient drug delivery. Such a concern is particularly important for osteosarcoma, the predominant form of bone cancer, and the largest cancer of its type in the paediatric age group. Pigment epithelium-derived factor (PEDF) is the most potent anti-angiogenic factor found endogenously in the body, with an increasing number of reports pointing to its direct antitumour activity. In this report, when a plasmid expressing PEDF (pPEDF) was encapsulated within two types of chitosan microparticles, anti-invasion and increased adhesion of the osteosarcoma cell line SaOS-2 was noted. Microparticles were formulated using two methods of complex coacervation and were approximately 400-600 nm in diameter. The plasmids were strongly attached to the particles which were polymorphic in shape as determined by electron microscopy. Preliminary experiments with the green fluorescent protein (GFP) reporter plasmid revealed that cells were efficiently transfected with the particles, with particles outlasting transfection with lipofectamine cationic liposomes at 5 days. In vivo, the better pPEDF microparticle resulted in a decrease in primary tumour growth, reduced bone lysis and reduced establishment of lung metastases in a clinically relevant orthotopic model of osteosarcoma. Thus, this new mode of localised gene delivery may hold promise for molecular therapy of osteosarcoma. PMID- 17408738 TI - Resilience, competence, and coping. PMID- 17408739 TI - Physicians' initial forensic impressions of hypothetical cases of pediatric traumatic brain injury. AB - OBJECTIVE: To describe physicians' initial forensic impressions of hypothetical cases of pediatric traumatic brain injury (TBI) and to compare the responses of pathologists and pediatricians. METHOD: A survey was administered to physicians who attended workshops on pediatric TBI; were members of two national internet list serves; and were members of the Section on Child Abuse and Neglect of the American Academy of Pediatrics (N=522) and the National Association of Medical Examiners (N=815). The survey included 16 hypothetical case scenarios depicting a pediatric TBI. Participants were asked to categorize their initial forensic impressions of each scenario on a seven-point scale from definitive unintentional to definitive inflicted injury. RESULTS: A total of 570 surveys were completed, including 465 of 1337 surveys (35%) distributed through the regular mail. In 8 of 16 hypothetical cases, a majority of respondents (range, 60-98%) and a majority of the more experienced respondents (range, 59-97%) categorized the hypothetical TBI as either unintentional or inflicted. Pathologists were less likely than the pediatric participants to view these scenarios as inflicted TBI. CONCLUSION: Certain features of TBI cases result in a majority of clinicians categorizing them as inflicted or intentional. In the absence of a confession of inflicted injury, witnessed or verifiable events, severe injuries without explanation or injuries that can be linked clearly to a perinatal period, clinicians were unable to agree on the etiology of the injury. Our results will be helpful to other researchers so that consistent, validated research definitions are used to classify pediatric TBI for research purposes. PMID- 17408740 TI - Characterization of the lactococcal group II intron target site in its native host. AB - The Lactococcus lactis group II intron (Ll.ltrB) retrohomes into the ltrB gene at high efficiency. To date, the critical DNA bases recognized in vivo by the Ll.ltrB ribonucleoprotein (RNP) have been exclusively elucidated in Escherichia coli. However, recent evidence indicates host-dependant differences in Ll.ltrB mobility, raising the possibility of limitations of the current model for RNP homing site recognition in the native L. lactis host. In this work, intron retargeting experiments in L. lactis have demonstrated that adherence to specific target site critical bases is not sufficient to predict success or failure of chromosomal invasion, as in E. coli. Accordingly, a quantitative real-time PCR (QPCR) assay was developed to test target site nucleotides previously demonstrated as critical for homing in E. coli, for relevance in its native host. This two-plasmid QPCR homing assay is highly sensitive and, unlike previous E. coli-based assays, resolves differential homing efficiencies in the absence of selection. As in E. coli, deviation from wild type at target site positions -23, 21, -20, -19, and +5 resulted in lower homing efficiencies in L. lactis. Furthermore, the same trends are observed when assaying select variants in Enterococcus faecalis. Our results suggest that these target site positions are critical in both E. coli and L. lactis. PMID- 17408741 TI - Characterization of Tn5386, a Tn916-related mobile element. AB - In recent work, we described excision of a large genomic region from Enterococcus faecium D344R resulting from the interaction of Tn916 and a related transposon designated Tn5386. In the present study, we present and analyze the complete sequence of Tn5386. Tn5386 is 29,451 bp in length. Fifteen of its 30 open reading frames are analogous to ORFs found in Tn916. Significant differences include a series of ORFs with homology to lantibiotic immunity genes in the same location where tetM is found in Tn916, insertion of a Group II intron and an ORF with similarities to previously described surface exposed collagen adhesion proteins. Our results indicate that Tn5386 falls within the Tn916 family of transposons, and in place of tetM encodes a novel region that may confer resistance to lantibiotics. PMID- 17408742 TI - Legal outcomes of all suspected neonaticides in Finland 1980-2000. AB - This nationwide study examined legal outcomes and possible psychiatric diagnoses of suspected cases of neonaticide. Neonaticide is commonly defined as the killing of a newborn on the day of its birth, and is considered to have not only a low prevalence but also a high level of concealed criminality. This hidden nature guided us to find out what the final legal outcomes of suspected neonaticide were. It was a comprehensive, retrospective, register-based study of all 44 cases of suspected neonaticide that occurred 1980-2000 as recorded by Statistics Finland. The 44 cases were ascribed to 40 suspects, three of whom died themselves during the offence. Twelve cases (27%) were eventually prosecuted and the accused convicted of neonaticide. Their mean sentence was 617 days (SD 216, range 300 1095 days). Fourteen offenders (35% of offenders) underwent a forensic psychiatric examination, out of which four (29%) were diagnosed with a psychotic disorder and 10 (71%) with a personality disorder. Six of the 14 women were not sentenced as criminally irresponsible and three of them were committed to involuntary hospital care. Nine cases (20% of cases) were still unsolved, and in eight (18%) cases the offence title had changed into something other than neonaticide. We concluded that since only 41% of suspected neonaticides completed the court process as neonaticides, the previous results from studies on neonaticide may present a subgroup of offenders, not the whole picture. Therefore, further discussion and research is needed to elucidate this perplexing, sad, and utterly redundant offence in modern society, to prevent it, and develop treatment programs for the offenders. PMID- 17408743 TI - Difference of T cell and B cell activation in two homologous proteins with similar antigenicity but great distinct immunogenicity. AB - The candidate particulate hepatitis E vaccine, HEV 239, has been shown to be an efficacious vaccine in primates, and clinical study to date shows it to be safe and immunogenic for humans. The antigenicity of HEV 239 is virtually identical to its N-terminal 26 amino acids truncated protein, E2, which is not particulate but soluble. However, HEV 239 is over 200 times more immunogenic than E2. In present study, several events underlying this dramatic immunogenicity difference have been addressed. (1) HEV 239 can efficiently evoke a vigorous and predominant T cell response while E2 cannot induce detectable T cell response; (2) the dominant T cell epitopes in HEV 239 are identified, and both are also contained integrally in E2; (3) priming mice with Th epitope peptide can partially rescue the weak immunogenicity of E2 in alum adjuvant and (4) HEV 239 but not E2 can induce significant antibody response in athymic mice, which indicates that HEV 239 can directly activate B cell more efficiently. These results contribute to a better understanding of the mechanisms involved in the significant high immunogenicity of particulate antigen and may provide knowledge for the rational design and development of future vaccines. PMID- 17408744 TI - Peptidomimics of antigen are present in variable region of heavy and light chains of anti-idiotypic antibody and function as surrogate antigen for perpetuation of immunological memory. AB - Peptide antigens composed of relevant B cell and T cell epitopes, capable of inducing protective immune response against the whole pathogen, are potentially safe, alternative vaccine antigens for prevention of wide range of diseases. Here, we show that short peptides derived from internal image sequences of anti idiotypic antibody (peptidomimics) can function as both B and T cell epitopes and perpetuate antigen specific immunological memory. We have sequenced the variable regions of heavy and light chains of the anti-idiotypic antibody specific to rinderpest virus hemagglutinin protein and predicted T cell epitopes in these sequences by an immuno-informatics approach. We have studied the interaction of these epitopes with MHC class I by in vitro assays and in silico analysis by molecular modeling of the idiopeptide-MHC complexes as well as antigen-derived peptide-MHC complexes. The functional capacity of anti-idiotypic antibody derived peptides to stimulate antigen specific T cells in vitro was tested. The ability of peptidomimics to proliferate the immune splenocytes in vitro was 10 times more when compared with that of a control peptide taken from the constant region of immunoglobulin. Similarly three- to fivefold more amounts of IL-2 and IFN-gamma were secreted by immune splenocytes in response to in vitro re-stimulation with peptidomimics. Further, we have provided evidence for the generation of antibodies against peptidomimics in memory response generated on antigen or anti idiotypic antibody immunizations. In summary, our experiments suggest that peptidomimics are generated in the body after antigen immunization and may have important roles in vivo in regulating antigen specific immunological memory. PMID- 17408745 TI - Munc18-2 regulates exocytotic membrane fusion positively interacting with syntaxin-3 in RBL-2H3 cells. AB - Recent studies have revealed that SNARE proteins are involved in exocytotic granular content release in mast cells as well as in neurotransmitter release in neural cells. However, the proteins that regulate the structure and activity of SNARE proteins in mast cells are not well understood. Munc18 is one such regulatory protein that plays a crucial role in neurotransmitter release. In this study, we investigated the role of Munc18 and its mechanism for regulating exocytotic release (degranulation) in rat basophilic leukemia cells (RBL-2H3). We obtained RBL-2H3 cells that express a low level of Munc18-2 and found that degranulation was remarkably inhibited in knockdown cells without any change in the expression level of syntaxins or Ca(2+) mobilization. We also observed the behavior of secretory granules in a single cell, and found no significant changes in their number and distribution in Munc18-2 knockdown cells. Using chimera proteins fused with fluorescent proteins, we demonstrated that Munc18-2 interacted with syntaxin-3, but not with syntaxin-4, in vivo. Interestingly, this interaction occurred not only on plasma membrane but also on secretory granules, suggesting that Munc18-2 may regulate granule-granule fusion as well as granule plasma membrane fusion. These observations suggest that Munc18-2 together with syntaxin-3 regulate degranulation positively during the process of membrane fusion between secretory granules and plasma membrane, rather than during processes that regulate the number or behavior of secretory granules. PMID- 17408746 TI - Effects of perinatal exposure to low-dose cadmium on thyroid hormone-related and sex hormone receptor gene expressions in brain of offspring. AB - Perinatal cadmium (Cd) exposure has been shown to alter behaviors and reduce learning ability of offspring. A few studies have shown that Cd reduced serum thyroid hormones (THs), which are important for brain development during the perinatal period. Brain specific genes, neurogranin (RC3) and myelin basic protein (BMP), are known to be regulated by TH through TH receptors (TR). It has been suggested that RC3 may play roles in memory and learning. In addition, Cd has been suggested to have estrogen-like activity. To evaluate the effects of perinatal low-dose exposure to Cd on thyroid hormone-related gene (RC3, TR-beta1, MBP, RAR-beta) and sex hormone receptor gene (ER-alpha, ER-beta and PgR) expressions in the brain and on behaviors of offspring, mice were administered with 10ppm Cd (from gestational day 1 to postnatal day 10) and/or 0.025% methimazole (MMI; anti-thyroid drug) (from gestational day 12 to postnatal day 10) in drinking water. Also, 0.1% MMI was administered as a positive control (high MMI group). RC3 mRNA expression was reduced in the female brain of combined exposure and high MMI groups and was negatively correlated with the activity in the open-field. ER-alpha, ER-beta and PgR mRNA expressions were decreased in male and female Cd, and female Cd+MMI groups, respectively; among these changes the reduced expression of PgR was opposite to estrogenic action. These results suggested that perinatal exposure to Cd disrupted the gene expressions of sex hormone receptors, which could not be considered to be a result of estrogenic action. Our study indicates that alteration in the gene expressions of RC3 and sex hormone receptors in the brain induced by perinatal Cd and MMI exposure might be one mechanism of developmental toxicity of Cd. PMID- 17408747 TI - Hanninen lecture: problems and prevention: research in developing countries and immigrant populations from developing countries. AB - The tradition of this excellent Triennial International Symposium, now in its Ninth gathering, has focused first on methods and later on effects in neurotoxicology. I respectfully suggest that it is time to add prevention to our agenda in order to focus not just on finding problems, but also on solving them. Otherwise our research runs the risk of making conditions worse for the participants because we inform them they have a problem but do not show them how to prevent that problem. Many barriers have been surmounted since the early days of this Symposium. Methods developed in industrialized nations have been modified and used in populations with limited education. Now, the primary barriers to effective collaborations are institutional. Perhaps the most challenging are companies that can bar a research study, or may welcome the opportunity to safeguard their workforce. A focus on prevention can increase the willingness of institutions, including companies, to accept and support research that examines neurotoxic effects of chemical exposures. This is because we are offering positive value, not simply uncovering a problem for them to fix. The cTRAIN software program is an illustration of a computer-based training method developed by neurotoxicologists that has been implemented in a variety of populations. This includes those from a developing country with limited education and those with college degrees. Training can provide prevention, can easily be appended to any neurotoxicology research study and is well within the skill set of the neurotoxicology community of scientists. PMID- 17408748 TI - Manganese induces cell swelling in cultured astrocytes. AB - Manganese in excess is neurotoxic and causes a CNS disorder that resembles Parkinson's disease (manganism). Manganese highly accumulates in astrocytes, which renders these cells more vulnerable to its toxicity. Consistent with this vulnerability, manganese has been shown to cause histopathological changes in astrocytes (Alzheimer type II change), generates oxidative stress and bring about mitochondrial dysfunction, including the induction of the mitochondrial permeability transition (mPT) in astrocytes. In addition to manganism, increased brain levels of manganese have been found in hepatic encephalopathy, a chronic neurological condition associated with liver dysfunction, wherein Alzheimer type II astrocytic changes are also observed. As low-grade brain edema, possibly secondary to astrocyte swelling, has been reported in hepatic encephalopathy, we hypothesized that manganese may contribute to such edema. We therefore exposed cultured astrocytes to manganese (Mn(3+)) acetate (25 and 50microM) for different time periods and examined for changes in cell volume. Manganese dose-dependently induced astrocyte swelling; such swelling was first observed at 12h (28%), which further increased (54%) at later time points (24-48h). Pretreatment of astrocyte cultures with antioxidants, including vitamin E, the spin trapping agent PBN, and the iron-chelating agent desferroximine, as well as the nitric oxide synthase inhibitor l-NAME, all significantly blocked (50-80%) astrocyte swelling caused by manganese, suggesting that oxidative/nitrosative stress is involved in the mechanism of such swelling. Cyclosporin A, an inhibitor of mPT also blocked (90%) manganese-induced astrocyte swelling. The data indicate that manganese exposure results in astrocyte swelling and such swelling, at least in part, may be caused by oxidative stress and/or mPT. Astrocyte swelling by manganese may represent an important aspect of manganese neurotoxicity, and may be a factor in low-grade brain edema associated with chronic hepatic encephalopathy. PMID- 17408749 TI - Cerium(IV)-mediated oxidation of flavonol with relevance to flavonol 2,4 dioxygenase. Direct evidence for spin delocalization in the flavonoxy radical. AB - The cerium(IV)-mediated oxidation of 3-hydroxy-4'-methylflavone (1) proceeds by H atom abstraction forming the flavonoxy radical (7), and the subsequent combination of its resonance forms leads to the 3-hydroxy-4'-methylflavone dehydro dimer (9). The above system serves as direct evidence for the intermediacy of the flavonoxy radical, its spin delocalization, and also indirect evidence for valence tautomerism as a key step on the substrate activation both in the quercetinase and its biomimic model system. PMID- 17408750 TI - Environmental factors and developmental outcomes in the lung. AB - The developing lung is highly susceptible to damage from exposure to environmental toxicants particularly due to the protracted maturation of the respiratory system, extending from the embryonic phase of development in utero through to adolescence. The functional organization of the lungs requires a coordinated ontogeny of critical developmental processes that include branching morphogenesis, cellular differentiation and proliferation, alveolarization, and maturation of the pulmonary immune, vasculature, and neural systems. Therefore, exposure to environmental pollutants during crucial periods of prenatal and/or postnatal development may determine the course of lung morphogenesis and maturation. Depending on the timing of exposure and pathobiological response of the affected tissue, exposure to environmental pollutants can potentially result in long-term alterations that affect the structure and function of the respiratory system. Besides an immature respiratory system at birth, children possess unique differences in their physiology and behavioral characteristics compared to adults that are believed to augment the vulnerability of their developing lungs to perturbations by environmental toxins. Furthermore, an interaction between genetic predisposition and increased opportunity for exposure to chemical and infectious disease increase the hazards and risks for infants and children. In this article, the evidence for perturbations of lung developmental processes by key ambient pollutants (environmental tobacco smoke [ETS], ozone, and particulate matter [PM]) are discussed in terms of biological factors that are intrinsic to infants and children and that influence exposure-related lung development and respiratory outcomes. PMID- 17408751 TI - Adenosinergic cardioprotection: multiple receptors, multiple pathways. AB - Adenosine, formed primarily via hydrolysis of 5'-AMP, has been historically dubbed a "retaliatory" metabolite due to enhanced local release and beneficial actions during cellular/metabolic stress. From a cardiovascular perspective, evidence indicates the adenosinergic system is essential in mediation of intrinsic protection (e.g., pre- and postconditioning) and determining myocardial resistance to insult. Modulation of adenosine and its receptors thus remains a promising, though as yet not well-realized, approach to amelioration of injury in ischemic-reperfused myocardium. Adenosine exerts effects through A(1), A(2A), A(2B), and A(3) adenosine receptor subtypes (A(1)AR, A(2A)AR, A(2B)AR, and A(3)AR), which are all expressed in myocardial and vascular cells, and couple to G proteins to trigger a range of responses (generally, but not always, beneficial). Adenosine can also enhance tolerance to injurious stimuli via receptor-independent metabolic effects. Given adenosines contribution to preconditioning, it is no surprise that postreceptor signaling typically mimics that associated with preconditioning. This involves activation/translocation of PKC, PI3 kinase, and MAPKs, with ultimate effects at the level of mitochondrial targets-the mitochondrial K(ATP) channel and/or the mitochondrial permeability transition pore (mPTP). Nonetheless, differences in cytoprotective signaling and actions of the different adenosine receptor subtypes have been recently revealed. Our understanding of adenosinergic cytoprotection continues to evolve, with roles for the A(2) subtypes emerging, together with evidence of essential receptor "cross-talk" in mediation of protection. This review focuses on current research into adenosine-mediated cardioprotection, highlighting recent findings which, together with a wealth of prior knowledge, may ultimately facilitate adenosinergic approaches to clinical cardiac protection. PMID- 17408752 TI - Anxiety disorders in a public mental health system: clinical characteristics and service use patterns. AB - BACKGROUND: Anxiety disorders are among the most common forms of psychiatric disorder, yet few investigations have examined the prevalence or service use of clients with anxiety disorders in the public mental health sector. METHODS: We examined demographics, clinical information, and service use in clients with anxiety disorders enrolled in San Diego County Adult and Older Adult Mental Health Services in fiscal 2002-2003. RESULTS: Almost 15% of the sample had a diagnosis of an anxiety disorder based on administrative billing data. Most anxiety disorder clients had additional psychiatric diagnoses, most commonly depression. Clients with both anxiety disorders and depression were more likely than those with anxiety or depression alone to use emergency psychiatric services and outpatient services than those with depression alone. Those with anxiety disorders alone used more outpatient services than those with depression alone. LIMITATION: Data were taken from an administrative database. CONCLUSIONS: Data indicate that anxiety disorders are not uncommon in public mental health settings and are associated with higher utilization of outpatient mental health services. PMID- 17408754 TI - The interaction of psychosocial adversity and biological risk in childhood aggression. AB - Childhood aggression has both biological and environmental underpinnings. However, the manner in which these factors interact to influence various types of aggression remains an important area of study. The current study examined the degree to which biological risk and psychosocial adversity, both alone and in combination, are associated with childhood aggression. Linear regression procedures were used to assess the extent to which biological risk status (low vs. high serotonergic responsivity, as measured by prolactin response to fenfluramine), magnitude of psychosocial risk, and the interaction of these factors predicted parent and teacher ratings of aggression and delinquency. After accounting for the independent contribution of biological and psychosocial risk, the interaction of biological and psychosocial risk was significantly associated with parent-rated aggression and marginally related to parent-rated delinquency. In contrast, no such interaction was observed for teacher-rated aggression. Findings suggest that individuals at biological risk for aggression may be particularly vulnerable to the impact of psychosocial adversity. PMID- 17408753 TI - The role of SOCS3 in modulating leukaemia inhibitory factor signalling during murine placental development. AB - Cytokines are an integral part of the adaptive and innate immune responses. The signalling pathways triggered by receptor engagement translate exposure to cytokine into a coordinated biological response. To contain these responses, the initiation, duration and magnitude of the signal is controlled at multiple levels. Suppressor of cytokine signalling (SOCS) proteins act in a negative feedback loop to inhibit signal transduction. Mice with a deletion of SOCS3 die at midgestion due to placental insufficiency. SOCS3-null placentae have increased numbers of mature trophoblast giant cells, disruption of the labyrinthine layer and a decrease in the spongiotrophoblast layer. Genetic crosses have revealed that the phenotype is due to dysregulation of signalling downstream of the leukaemia inhibitory factor (LIF) receptor alpha (LIFRalpha) and that the ligand responsible for this, LIF, is produced by embryonic tissues and acts in a paracrine fashion. These observations highlight the role of LIF as an extrinsic factor regulating trophoblast differentiation in vivo. The creation of mice with conditional deletion of SOCS3 in different tissues has also uncovered critical roles for SOCS3 in the regulation of IL-6, G-CSF and leptin signalling. PMID- 17408755 TI - Chronic foot shock induces hyperactive behaviors and accompanying pro- and anti inflammatory responses in mice. AB - Behavioral and accompanying physiological and immunological changes were investigated at various times during chronic irregular mild foot shock (CMFS) in adult male BALB/c mice. CMFS induced a significant hyperlocomotor activity in a familiar environment as well as increased consumption of chocolate milk (a favored drink) throughout the 5-week stress period. Unlike other chronic stress models, CMFS did not induce depressive-like behaviors. Hyperactivity was associated with transient elevations of pro-inflammatory cytokines (TNFalpha and IL-1beta) and IL-2 and more sustained (IL-10) or later (arginase activity) elevations in anti-inflammatory mediators in the spleen (serum levels below levels of detection) suggesting a transition from a pro-inflammatory state to an anti-inflammatory state during CMFS. Similar increases in brain levels of IL-2 and arginase activity were also detected and may contribute to CMFS-induced hyperactivity as both of these mediators have been shown to induce hyperactivity. To our knowledge, this is the first time that increased arginase activity has been documented during a stress paradigm. Altogether, the data indicate that CMFS induces behavioral changes distinct from other chronic stress models. CMFS is associated with multiple dynamic immunological changes, suggesting involvement of multiple factors in chronic stress-induced behavioral changes. PMID- 17408756 TI - Tracheobronchial foreign bodies in children. Experience at ORL clinic Tuzla, 1954 2004. AB - OBJECTIVE: To determine the clinical characteristics and the results of bronchoscopic treatment of children due to foreign body aspiration in a university hospital. SETTING: Department of Otorhinolaryngology and Maxillofacial Surgery, University Clinical Center Tuzla, Bosnia and Hercegovina. METHOD: The analysis of the cases of aspirated foreign bodies within the period from January 1954 to December 2004. The analyzed patients were the children up to 14 years of age. All cases underwent the bronchoscopy. Each patient was analyzed for age, sex, nature and location of the foreign body, results of bronchoscopic removal, complications of bronchoscopy and presence of foreign bodies in the airways. RESULTS: Six hundred and sixty-two children who underwent bronchoscopy for removal of foreign body in the airways were evaluated. From evaluated children 66.8% were boys, ages ranging from 9 months to 14 years. Foreign bodies were more frequent in children under 3 years of age (65.2%). Most of the foreign bodies removed were organic (87.1%) and more frequently found in the right bronchial tree (53%). CONCLUSIONS: More attention is necessary to the prevention of aspirations. Prevention of aspiration of foreign bodies is better than cure. Public awareness through mass media needs attention to prevent foreign body inhalation. PMID- 17408757 TI - Endoscopic management of recurrent congenital tracheoesophageal fistula: a review of techniques and results. AB - OBJECTIVE: Recurrent tracheoesophageal fistula (TEF) is a difficult problem in both diagnosis and management. Revision open repair with thoracotomy is challenging and has a significant associated morbidity. Because of the technical difficulty and the substantial morbidity, several authors have suggested and implemented endoscopic management. This paper reviews and describes the endoscopic techniques and management of recurrent TEF. METHODS: This retrospective study evaluates three patients who underwent endoscopic (bronchoscopic) management of recurrent TEF at Children's Memorial Hospital in Chicago, Illinois. Median follow-up is 48 months. A current literature review is presented with a synthesis of the data on techniques and results. RESULTS: Two of the three patients had successful bronchoscopic closure using fibrin adhesive. The mean number of procedures required was two. Including these patients, 62 patients with endoscopic closure of congenital TEF have been reported in the literature. The overall success rate is 60% with a mean of 2.1 procedures per patient. Of the obliterating agents described, tissue adhesive and fibrin adhesive have been employed most frequently with success rates of 48 and 55%, respectively. CONCLUSION: Endoscopic repair of recurrent TEF is an effective and safe alternative to second thoracotomy and open surgical repair. More than one endoscopic procedure is usually necessary for successful closure. PMID- 17408758 TI - D1 and D2 dopamine-receptor modulation of striatal glutamatergic signaling in striatal medium spiny neurons. AB - Dopamine shapes a wide variety of psychomotor functions. This is mainly accomplished by modulating cortical and thalamic glutamatergic signals impinging upon principal medium spiny neurons (MSNs) of the striatum. Several lines of evidence suggest that dopamine D1 receptor signaling enhances dendritic excitability and glutamatergic signaling in striatonigral MSNs, whereas D2 receptor signaling exerts the opposite effect in striatopallidal MSNs. The functional antagonism between these two major striatal dopamine receptors extends to the regulation of synaptic plasticity. Recent studies, using transgenic mice in which cells express D1 and D2 receptors, have uncovered unappreciated differences between MSNs that shape glutamatergic signaling and the influence of DA on synaptic plasticity. These studies have also shown that long-term alterations in dopamine signaling produce profound and cell-type-specific reshaping of corticostriatal connectivity and function. PMID- 17408759 TI - Dopamine neuron systems in the brain: an update. AB - The basic organization of the catecholamine-containing neuronal systems and their axonal projections in the brain was initially worked out using classical histofluorescence techniques during the 1960s and 1970s. The introduction of more versatile immunohistochemical methods, along with a range of highly sensitive tract-tracing techniques, has provided a progressively more detailed picture, making the dopamine system one of the best known, and most completely mapped, neurotransmitter systems in the brain. The purpose of the present review is to summarize our current knowledge of the diversity and neurochemical features of the nine dopamine-containing neuronal cell groups in the mammalian brain, their distinctive cellular properties, and their ability to regulate their dopaminergic transmitter machinery in response to altered functional demands and aging. PMID- 17408760 TI - Synergistic inhibition of HIV-1 infection by combinations of soluble polyanions with other potential microbicides. AB - Several polyanionic compounds with potential for use as topically applied microbicides to prevent HIV-1 sexual transmission, such as PRO 2000, are currently in phase III clinical efficacy trials. Microbicidal formulations may well comprise combinations of inhibitors to increase potency, reduce dose and minimize problems of HIV-1 resistance. We have therefore evaluated in vitro, the anti-HIV-1 activity of two leading polyanionic microbicides combined with other antiretroviral agents with microbicidal potential. Dextran sulfate (DS) and PRO 2000 were combined with the neutralizing antibody IgG1b12, the peptide-based fusion inhibitor T20, the CCR5 antagonist TAK779 and the cyanobacterial protein cyanovirin-N. Anti-HIV-1 activity was assessed in a single cycle replication assay using pseudoviruses carrying a luciferase reporter gene and the envelope glycoproteins from HIV-1 isolates JR-FL (R5) and HxB2 (X4), against both immortalized and primary CD4+ cell targets. The data were analyzed for synergy using Calcusyn software. Results indicate that PRO 2000 and DS can act synergistically with most inhibitors tested, although the degree of synergy depends on inhibitor concentration and combination. These data provide a rational basis for testing of microbicide combinations in vivo. PMID- 17408761 TI - An approach to evaluate the ability of rats to discriminate different levels of illumination in the plus maze test: effects of scopolamine. AB - Previous research has shown that the visual system is important for rats to establish the arm preference in the elevated plus maze (EPM), an animal model of anxiety. This study aims at evaluating whether a gradient of illumination between the enclosed arms of the maze (E/E(DeltaLux)) could be a reliable approach to detect drugs-induced harmful effect on visual discrimination of rats. Four EPM configurations with different E/E(DeltaLux) (8, 41 and 85lx) were used to demonstrate that as E/E(DeltaLux) increases, rats avoid to explore the light enclosed arm, which characterizes the animal ability to discriminate the most illuminated area within the protected environment of the maze. The establishment of either 41 or 85 E/E(DeltaLux) failed to alter the traditional spatial-temporal variables in the EPM. In addition, systemic treatment with midazolam (MDZ; 1.0mgkg(-1), a classical anxiolytic) induced anxiolysis in rats tested in 41 and 85 E/E(DeltaLux) EPM, with no change in the visual discrimination, when evaluated by the level of light enclosed arm exploration. Systemic treatment with scopolamine (SCP; 1.0, 2.0 and 8.0mgkg(-1)), a drug endowed with harmful properties upon the visual system, did not change either the open arm avoidance or the visual discrimination at the low doses, but induced increased light enclosed arm (visual discrimination deficit) and open arm exploration (anxiolytic like effect) at a higher dose. We propose that the incorporation of an E/E(DeltaLux) in the EPM may reinforce the predict validity of the test since it enables to evaluate whether a visual discrimination deficit can be confounded with an anxiolytic-like effect, thus establishing a false positive detection. PMID- 17408762 TI - Pre- and postnatal FGF-2 both facilitate recovery and alter cortical morphology following early medial prefrontal cortical injury. AB - Rats with either no treatment or administration of exogenous basic fibroblast growth factor (FGF-2) received bilateral medial prefrontal cortical (mPFC) aspiration or sham lesions at postnatal day 3 (P3). FGF-2 was administered either prenatally at embryonic day 15.5 (PreFGF) or, postnatally (PostFGF) for 7 consecutive days beginning 1 day following surgery. As adults, animals were tested behaviorally at spatial navigation (Morris water task), and skilled reaching (Whishaw tray reaching task). Early lesions of the mPFC produced a significant reduction in both brain weight and cortical thickness in adulthood. Behaviorally, mPFC lesions resulted in deficits in the water maze and reaching task. Both pre- and postnatal FGF-2 facilitated recovery in the spatial navigation task. In contrast, FGF-2 was only effective in reducing the deficits in skilled forelimb movements when the FGF was given postnatal (i.e., postsurgery). Prenatal FGF-2 increased brain weight in the lesion animals, whereas postnatal FGF-2 increased cortical thickness in the lesion animals. It thus appears that FGF-2 can facilitate recovery from perinatal cortical injury, whether it is given during the period of neurogeneration (prenatally) or after the injury, although the mechanism of action is likely different for the pre- and postnatal administration. PMID- 17408763 TI - Repeated measurements of learned irrelevance by a novel within-subject paradigm in humans. AB - Learned irrelevance (LIrr) refers to the retardation of classical conditioning following preexposure of the to-be-associated stimuli. Healthy volunteers have been tested on three occasions with a new LIrr paradigm avoiding methodological problems which afflict traditional paradigms. A significant LIrr effect was demonstrated on each occasion. Thus, the new paradigm enables repeated measurements of LIrr and might be useful in evaluating long-term effects of medication in psychiatric disorders exhibiting aberrant LIrr. PMID- 17408764 TI - Diabetic type II Goto-Kakizaki rats show progressively decreasing exploratory activity and learning impairments in fixed and progressive ratios of a lever press task. AB - Learning and memory impairments associated with diabetes have been reproduced in rodent models of diabetes type I, but few studies have been performed in spontaneously type II diabetic rodents. The study of type II diabetic rats such as the Goto-Kakizaki (GK) rat is of advantage when characterizing the development of cognitive impairments specifically caused by the progression of the disease and not by its treatment. We thus hypothesized that GK rats might display learning impairments when compared to non-diabetic Wistar rats. In the present study, we employed a lever-press task, a behavioural paradigm which allows the study of response-reinforcement learning, discrimination of a rewarding lever (using a two-choice positional discrimination task), and the ability to increase operant behaviour when requirements for reward increase (using a progressive ratio [PR]). In parallel, locomotor activity was compared between strains to assess exploratory activity and behavioural habituation to a novel environment. Diabetic GK rats made significantly less lever-presses with increasing fixed ratios and, throughout the sessions, a trend for increased selection errors was observed in these animals. In addition, a significant reduction in the maximum number of lever-presses made by GK rats was observed during the PR sessions. Locomotor activity of GK rats was higher on the first day of exploration but significantly decreased with familiarization to the environment. The present results suggest that the diabetic-like symptomatology in GK rats led to a reduction of exploratory activity and of lever-pressing during fixed and progressive ratio schedules, likely caused by learning impairments. PMID- 17408765 TI - Nuclear localization of an actin-related protein (ORF LmjF21.0230) in Leishmania+. PMID- 17408766 TI - Highly efficient stable transformation of bloodstream forms of Trypanosoma brucei. PMID- 17408767 TI - Thromboembolism in takotsubo syndrome: a case report. PMID- 17408768 TI - Infective endocarditis of aortic valve during pregnancy: a case report. AB - Infective endocarditis during pregnancy is uncommon but very serious. A 31-year old woman in the 36th week of second pregnancy was admitted to a hospital because of fever, weakness, chest pain, painful skin over her right leg and dyspnea. Transthoracic echocardiography showed aortic valve vegetation and severe aortic regurgitation. Transesophageal echocardiography revealed a 18 mmx6 mm mobile vegetation, attached to the right coronary cusp. Emergency cesarean section followed with a delivery of a healthy baby. Cardiopulmonary bypass with subsequent aortic replacement with bioprosthesis was initiated immediately after cesarean section. Early echocardiographic examination and 6 months after surgery revealed normal function of aortic valve bioprosthesis and normal LV function. Clinical recognition and early echocardiographic diagnosis followed urgent simultaneous cesarean section and aortic valve replacement was lifesaving for both mother and fetus. PMID- 17408769 TI - A successfully novel ICD implantation and medical treatment in a child with LQT syndrome and self-limiting ventricular fibrillation. AB - Beta-blocker is the first line drug therapy for congenital long QT syndrome. However, in some children this drug is ineffective. In a non-responder patient, Shimizu et al. used Mexiletine to suppress the ventricular arrhythmias, obtaining a good result. In the high risk patient, the ICD is necessary. However the implantation of a device in small children can have technical problems. We report a case of a child affected by long QT syndrome with recurrent episodes of syncope due to self-limiting torsade de point/ventricular fibrillation, successfully treated by an association of mexiletin and propanolol, and in whom an ICD was implanted with a new subcutaneous approach. PMID- 17408770 TI - Sulpiride induced torsade de pointes. AB - Antipsychotic agents are known to be associated with a long QT interval and torsade de pointes. We report a 69 year old female who suffered from a syncopal attack at a psychiatric hospital and was referred to our center. Torsade de pointes with a long QT interval (QTc=680 ms) was observed on the 12 lead ECG in the emergency department and intensive care unit. A careful drug history revealed that sulpiride was the culprit agent. After stopping the medication, the QT interval returned to normal (420 ms). The patients taking sulpiride should be closely monitored, especially when it is used in combination with other antidepressant agents. PMID- 17408771 TI - Uncommon presentation of asymmetric hypertrophic cardiomyopathy or something else. PMID- 17408772 TI - Unusual electrocardiographic presentation of typical atrioventricular nodal re entrant tachycardia. AB - The present case report illustrates an atypical electrocardiographic presentation of typical atrioventricular nodal re-entrant tachycardia. Persistent 2:1 AV block during tachycardia is evident and an spontaneous ventricular premature beat during tachycardia resulted in 1:1 AV conduction. PMID- 17408773 TI - Dexamethasone-eluting stents for the prevention of in-stent restenosis: evidence for a differential effect in insulin-dependent and non-insulin-dependent diabetic patients. AB - Diabetes mellitus (DM) is a strong predictor of in-stent restenosis. This may be due to a higher level of vascular inflammation. We hypothesized that diabetic patients will benefit from dexamethasone-eluting stents, since local inflammation and consequently neointimal growth are suppressed and no systemic side effects will occur. METHODS: 21 consecutive patients with DM with 32 lesions were treated with dexamethasone-eluting stents. Excluded were patients with triple vessel disease, bifurcation lesions, previous revascularization of the culprit vessel, and reference diameter smaller than 2.5 or larger than 3.75 mm. MACE (death, myocardial infarction, and revascularization) was counted at 12 months. At 6 months, angiographic follow-up was performed. RESULTS: Of the patients, 38% had insulin-dependent DM. Lesion type was type A/B1 in 56% and B2/C in 44%. Lesion length was 15.7+/-8.4 mm and the reference diameter was 2.83+/-0.53 mm. Event free survival at 12 months was 62%. Any revascularization procedure was performed in 33% and target lesion revascularization in 24% of the patients. At 6 months in stent late loss was 1.07+/-0.64 mm. Binary restenosis occurred in 28.1% of the lesions. The event-free survival in insulin-dependent DM was worse compared to non-insulin-dependent DM (92.1 vs. 37.8%; p<0.01). Patients with insulin dependent DM had higher in-stent late loss compared to non-insulin-dependent DM patients (1.44+/-0.83 vs. 0.83+/-0.51 mm; p<0.01). CONCLUSION: Treatment with dexamethasone-eluting stents in patients with DM is associated with a relatively high restenosis rate. Our data suggest a differential effect of dexamethasone eluting stents in insulin-dependent compared to non-insulin-dependent DM. PMID- 17408774 TI - Protracted presyncope and profound bradycardia due to lithium toxicity. PMID- 17408775 TI - The increase in P-wave dispersion is associated with the duration of disease in patients with Behcet's disease. AB - AIM: P-wave dispersion (PD) has been reported to be associated with inhomogeneous and discontinuous propagation of sinus impulses. Behcet's disease (BD) is a chronic multi-system disease presenting with recurrent oral and genital ulceration, and relapsing uveitis. Left ventricular diastolic dysfunction, ventricular arrhythmia and sudden cardiac death have been documented in Behcet's disease. The aim of this study was to evaluate myocardial involvement noninvasively in patients with BD by measuring PD. METHODS: Study population included 29 patients with BD (group I), (16 males, mean age: 60.4+/-10.1 years), and 45 normal (group II), (23 males, mean age: 61.2+/-12.0 years). The maximum and minimum P-wave duration (Pmax and Pmin) and PD were measured from the 12-lead surface electrocardiogram. Echocardiographic examination was also performed. RESULTS: There was no statistically significant difference between two groups in respect to age, gender, hypertension, diabetes mellitus, hyperlipidemia and cigarette smoking (p>0.05). Pmax and PD of patients with BD were found to be significantly higher than those of control subjects (116.2+/-14.0 ms vs. 102.0+/ 16.5 and 49.3+/-12.7 ms vs. 29.3+/-8.5 ms, respectively p<0.0001). Moreover, we found a significant positive correlation between PD with BD duration (r=0.78, p<0.0001). CONCLUSION: Pmax and PD are found to be greater in patients with BD than in controls and are related with the duration of the disease. PMID- 17408776 TI - Right ventricle injury during acute pulmonary embolism leads to its remodeling. AB - Right ventricular (RV) overload and hypoxia in acute pulmonary embolism (APE) may lead to RV myocardium injury reflected by elevated cardiac troponin levels. We studied 26 patients aged 57.2+/-17.8 years with first episode of APE. On admission troponin T (TnT) was measured. Transthoracic echocardiography was performed after 6 months of anticoagulation. Myocardial injury (TnT > or =0.03 ng/ml) was observed in 8 (30.8%) patients at the diagnosis. At follow up RV diastolic area tended to be larger in group with myocardial injury (25.0 (20.8 38.6) vs 18.4 (17.7-23.3) cm(2), p=0.06). Tricuspid annulus systolic velocity at tissue Doppler was lower in group with myocardial injury (0.12 (0.11-0.13) vs 0.15 (0.13-0.21) m/s, p=0.04), while no such a relationship was found for mitral annulus systolic velocity. TnT concentration correlated with RV diastolic area (r=0.61) and tricuspid annulus systolic velocity (r=-0.58) although not significantly (p=0.08 and p=0.09. respectively). Our data suggest that RV injury in acute phase of PE may lead to its remodeling. PMID- 17408777 TI - Reproducibility and safety of the incremental shuttle walking test for cardiac rehabilitation. AB - The incremental shuttle walking test (ISWT) is recommended to assess the exercise capacity of patients attending cardiac rehabilitation (CR) as a cost-effective alternative to the treadmill test. We used the Bland Altman limits of agreement method on 353 paired ISWTs to explore whether the recommended practice test is necessary. The mean increase in distance from the practice to the second walk was 29.5 m (95% CI 23.0, 36.0) p<0.001, confirming the need for the practice walk when undertaking the ISWT. PMID- 17408778 TI - Cardiovascular prognostic value of vascular involvement in Behcet's disease. AB - We measured carotid artery intima-media thickness (IMT), brachial artery flow mediated dilation (FMD), and coronary flow reserve (CFR) in 38 Behcet's disease (BD) patients without vascular involvement, 15 BD patients with vascular involvement, and 35 control subjects. BD patients with and without vascular involvement were similar regarding carotid IMT (0.52+/-0.14; 0.51+/-0.09; 0.46+/ 0.09; P=0.051), and brachial FMD (16.3+/-6.8; 16.5+/-9.1). CFR values were significantly impaired in BD patients compared to the controls; however the BD patients with and without vascular involvement were similar regarding CFR (2.66+/ 0.45; 2.62+/-0.48; 2.91+/-0.53, P=0.007 with and without vascular involvement and the controls, respectively). Endothelial and coronary microvascular functions are compromised in BD patients without vascular involvement as remarkably as that in patients with vascular involvement. PMID- 17408779 TI - Rheumatic mitral valve stenosis is associated with impaired flow-mediated dilatation. AB - It has been demonstrated that rheumatic mitral valve stenosis (RMVS) is associated with an increase in markers of endothelial dysfunction. It is not known whether this association indicates an impairment of flow-mediated dilatation (FMD) of the vascular endothelium. Thirty patients with RMVS and 30 healthy subjects were studied. FMD in patients with RMVS was significantly smaller than in healthy controls (11.9+/-0.4% vs 15.4+/-0.70%, p=0.003). The absolute change in brachial artery diameter in patients with RMVS was also significantly smaller than in healthy subjects (0.42+/-0.26 mm vs 0.64+/-0.32 mm, p<0.001). These findings suggest that vascular endothelial function is altered in patients with RMVS. PMID- 17408780 TI - Causes of ineligibility in randomized controlled trials and long-term mortality in patients with non-ST-segment elevation acute coronary syndromes. AB - PURPOSE: To determine the long-term mortality of patients with non-ST-segment elevation acute coronary syndromes (NSTEACS) that are eligible versus those not eligible in randomized controlled trials (RCT), and how each exclusion criteria is associated with outcome. METHODS: Common causes of exclusion in six published RCT on intravenous antithrombotic therapy were prospectively assessed in a cohort of 452 consecutive patients with NSTEACS that were followed for up to 3 years. RESULTS: Forty-one percent of patients had one or more exclusion criteria establishing the ineligible group. These patients were older, more likely to have coronary risk factors, ischemic ECG changes, heart failure at admission, higher creatinine levels and a lower ejection fraction than eligible patients. There were no differences between both groups in the antithrombotic treatment received or in the performance of revascularization procedures during hospitalization or in the prescription of antiplatelet treatment and beta-blockers at discharge. Cumulative 3-year mortality rate was 25% in ineligible patients compared to 9% in eligible patients (p<0.001). The hazard ratio (HR) of mortality was of 9.1 (95% CI: 4.5-18.7) for severe renal dysfunction; 6.0 (3.3-11.4) for concomitant non vascular diseases; 3.0 (1.6-5.5) for contraindications to anticoagulation; 2.5 (1.1-5.7) for heart failure; and 2.3 (1.1-4.6) for prior cerebrovascular disease. After adjusting for baseline differences, ineligible patients had a HR of total mortality of 1.88 (1.04-3.38), and of cardiac mortality of 2 (1.01-3.98). CONCLUSION: Patients with NSTEACS who are ineligible in RCT have a higher risk profile and a two-fold adjusted long-term mortality than eligible patients, especially those with comorbid conditions and those with contraindications to anticoagulation. PMID- 17408781 TI - Optimal fluoroscopic view selection for percutaneous coronary intervention by multislice computed tomography. AB - We present 2 cases of patients with stenotic and occlusive coronary lesions, which were detected by multislice computed tomography (MSCT) coronary angiography and treated with percutaneous coronary intervention (PCI) using CT-oriented optimal fluoroscopic views. Preprocedural MSCT allowed us to select the optimal fluoroscopic angle to visualize the target lesions, which provided least amount of foreshortening and minimal overlap of side branches during the PCI procedures. Given its three-dimensional nature, MSCT provides additional anatomical information in the evaluation of complex coronary lesions prior to PCI. PMID- 17408782 TI - A rare case of multiple right heart myxomas. AB - Primary tumours of the heart are rare. Myxomas are the most common primary cardiac tumours with an estimated incidence of 0.5 per million population per year. Myxomas are most commonly found in left atrium and are solitary in more than 90% cases. Familial myxomas are rarer and tend to be multiple. We report a rare case of multiple myxomas in the right heart which occurred sporadically, presenting with an episode of pre-syncope. PMID- 17408783 TI - A congenital absence of the pericardium diagnosed by echocardiography. AB - A 29-year-old man visited our office after experiencing palpitations at night. The electrocardiogram revealed premature ventricular contractions (PVCs), and he was admitted to our hospital for further tests. The echocardiography revealed paradoxical motion of the interventricular septum. After considering results from a previous cardiac catheter test, we suspected that the patient may have a congenital absence of the pericardium. A CT scan and magnetic resonance imaging were inconclusive, but through echocardiography and ECG performed during three positional changes we diagnosed a congenital absence of the pericardium. PMID- 17408784 TI - Anthropometric correlates of blood pressure in normotensive Pakistani subjects. AB - Obesity and hypertension are two major inter-related cardiovascular risk factors. Decrease in adiposity is one of the most effective preventive measures not only in decreasing the overall cardiovascular risk but also the blood pressure. This cross-sectional study measured the effect of various measures of adiposity on blood pressure in normal healthy subjects of Pakistani origin. 400 normotensive subjects (247 males and 153 females) were included in this study. Along with data on co-morbid conditions, two blood pressure readings and several anthropometric measurements were recorded. Age and gender specific analysis was done. Following the WHO cutoffs for Asians, about 52% of our sample population was found to be overweight or obese. Age was not associated with blood pressure indices in males; however it was strongly associated with all blood pressure indices in females. Greater Body Mass Index (BMI), Waist Circumference (WC) and Waist to Height Ratio (WHTR) were associated with higher Systolic and Diastolic Blood pressure. Increasing age was also associated with higher levels of BMI, WC and WHTR. Anthropometric variables however, were more strongly associated with blood pressure indices than age in this sample population. In conclusion, we found WC and WHTR to be strongly associated with blood pressure indices in normotensive Pakistani males. PMID- 17408785 TI - Tako-Tsubo cardiomyopathy characteristics in long-term follow-up. AB - Tako-Tsubo cardiomyopathy (TTC) is described as left ventricular (LV) dysfunction with the phenomenon of "apical ballooning", rapidly resolving, without coronary artery stenoses. METHODS: Fifteen patients with TTC and transthoracic echocardiography (TTE) at their admission, were reviewed (2001 to 2006). Follow up (F/U) TTE was performed in varying intervals. To compare diameters of posterior wall (PW), interventricular septum (IVS), left atrium (LA), LV in end diastole (LVED) and LV in end-systole (LVES) and valve insufficiencies, patients with comparable F/U are selected. RESULTS: Fourteen patients were female (mean age 69.6 years). Angiography demonstrated LV systolic dysfunction with mean ejection fraction (EF) of 31.3%. In the acute-phase (day 0 to day 3), TTE showed a mean EF of 35.7%, not significantly different from EF obtained in angiography. Short-term F/U was performed in 9 patients after median time-interval of 20 days with an increase to a mean EF of 58.8%. F/U in 2006 has been performed in 10 patients (median time-interval 18.7 months) and showed normal EF. No significant difference in diameters of LA, LVED and LVES could be obtained comparing baseline and long-term data (p=0.493, p=0.790 and p=0.275). PW and IVS were significantly thicker at baseline compared to TTE > or = day 62 of F/U (p=0.003 and p=0.026). At baseline mitral valve insufficiency (MI) was mild in 50.0% and moderate in 12.5%, mild and moderate tricuspid valve insufficiency (TI) was recognized in 50% (25% respectively). MI and TI were regredient in F/U. In three patients an intraventricular systolic flow acceleration could be detected in the acute phase. CONCLUSIONS: Characteristics of TTC, besides transient LV apical ballooning are also a significant change in LV wall thickness and reversible valve insufficiencies. PMID- 17408786 TI - Radiation dose exposure in multislice computed tomography of the coronaries in comparison with conventional coronary angiography. AB - BACKGROUND: Radiation dose exposure is increased in multislice spiral computed tomography (MSCT) compared to conventional coronary angiography (CXA). METHODS: Retrospective data analysis of 56 patients (66+/-8 years, 49 males, body surface area 1.98+/-0.18 m(2), heart rate 64+/-11 bpm) who underwent MSCT and CXA was performed (MSCT: 16-slice scanner, rotation time 0.375 s, 120 kV, ECG-pulsing; CXA: current technique system build in 2003). Ten patients with bypass grafts underwent bypass angiography in CXA and MSCT. To compare the radiation doses of both investigations, the effective dose (ED) was chosen as the analysis variable. RESULTS: The mean ED for MSCT was 9.76+/-1.84 mSv (n=46) for patients without bypass grafts; with calcium scoring the mean ED was 12.46+/-2.23 mSv (n=46). In comparison, the mean ED of CXA was 2.60+/-1.27 mSv (n=46) for patients without bypass grafts; with bypass grafts (n=10) the mean ED for MSCT was 12.95+/-1.75 mSv, for CXA of 6.27+/-4.04 mSv, respectively. In MSCT heart rates of 60 bpm (8.86+/-1.24 mSv versus 10.53+/-1.86 mSv). CONCLUSIONS: MSCT is still associated with a higher radiation dose exposure than CXA. The radiation dose relation is more favorable for MSCT than for CXA in patients with bypass grafts in comparison to patients without bypass grafts. This study emphasizes the importance of dose reduction techniques. PMID- 17408787 TI - Transient syncope, left bundle branch block and first degree atrioventricular block after "pill-in-the-pocket" administration. PMID- 17408788 TI - Renal cell carcinoma in the left ventricular outflow tract. PMID- 17408789 TI - A fluorescence assay to determine the viable biomass of microcosm dental plaque biofilms. AB - Dental plaque bacteria form complex and robust cell aggregates which cannot be counted accurately using epifluorescence microscopy. This causes a significant problem for quantifying their viability. The aim of the investigation was to develop a fluorescence assay to quantify the viable biomass of dental plaque biofilms. Using an artificial mouth system, microcosm plaques were grown under a range of fluoride and mineralizing conditions, and were treated with the oral antiseptics chlorhexidine (CHX) and Listerine. Plaques were harvested, made into suspension and stained in microtitre plates with a di-chromatic fluorescent stain (Live/Dead BacLight). The percentage of viable biomass was calculated from the regression data generated from a viability standard. The standard was constructed using different proportions of viable (green fluorescence) and non-viable (red fluorescence) plaque bacteria, and growth conditions for optimizing green fluorescence were investigated. The results from the assay showed that fluoride at 1000 and 3000 ppm promoted plaque viability by at least 15%, from approximately 45 to 60%, and at 5000 ppm to approximately 87% (P<0.05). Plaques treated with Listerine and CHX from d 0 yielded insufficient biomass to be tested for viability, however 14 d post-treatment, viability was comparable to untreated plaques (approximately 55%, P>0.05). Treatment with Listerine and CHX from d 3 reduced biomass but not viability. Development of this assay enabled viability of plaque bacteria which cannot be resolved with epifluorescence microscopy to be evaluated. It offers a rapid alternative to epifluorescence microscopy and could be applied to nonoral bacteria. PMID- 17408790 TI - Improved 16S rRNA-targeted probe set for analysis of sulfate-reducing bacteria by fluorescence in situ hybridization. AB - An updated dataset of in silico specificities for 54 previously published 16S rRNA-targeted oligonucleotides was assembled to provide guidance for reliable fluorescence in situ hybridization (FISH) analysis of sulfate-reducing bacteria. Additionally, six new FISH probes were developed for major deltaproteobacterial taxa, including a probe trio targeting most Deltaproteobacteria and Gemmatimonadetes. PMID- 17408791 TI - Effect of human embryonic stem cell-derived neuronal precursor cell transplantation into the cerebral infarct model of rat with exercise. AB - We analyzed the therapeutic effect of the transplantation of the human embryonic stem cell (NIH Code: MB01)-derived neuronal precursor (hES-NP) cell and post ischemic exercise in rats with the middle cerebral artery (MCA) infarct model. A cortical infarct was induced in 20 adult Sprague-Dawley rats by occlusion and reperfusion of the MCA. The rats were divided into four groups: hES-NP cell transplantation and exercise, transplantation only, exercise only, and Sham operated with no exercise. In the cell-transplanted group, hES-NP cells were transplanted by stereotactic inoculation into the ipsilateral basal ganglia 7 days after infarct. We evaluated the clinical recovery of deficit, the size of infarct and the survival, migration, and differentiation of the transplanted cells. The transplanted hES-NP cells survived robustly in the ischemic brains 3 weeks post transplant. The majority of migrating cells in the ischemic rats had a neuronal phenotype. The clinical scores of all of the experimental groups were better than those of the Sham-operated group. Whereas the exercise-only group showed continuous clinical improvement, the cell-transplanted groups manifested less improvement than the exercise-only group. Moreover, the cell-transplanted groups did not differ in clinical improvement according to postinfarct-exercise or not. The infarct size was significantly reduced in both the cell-transplanted groups and the post-ischemic exercise group, compared with the Sham-operated group; however, the reduction of infarct size was most prominent in the exercise only group. In our study, the inoculated site of the basal ganglia showed some damage induced by inoculation, such as loss of neuroglial cells, reactive gliosis and microcalcification, which was found in the Sham-operated group as well, and yet no inoculation-site injury has ever been reported. Our study revealed that stem cell transplantation can have a positive effect on behavioral recovery and reduction of infarct size, but the effect shown was no better than the effect of the exercise, which finding reconfirmed the importance of post-infarct rehabilitation. In addition, it was found that cell inoculation should be replaced by a noninvasive procedure. PMID- 17408792 TI - Different patterns of biofilm formation in Staphylococcus aureus under food related stress conditions. AB - Staphylococcus aureus and its biofilm formation are recognized as a serious clinical problem. S. aureus is also a food borne pathogen, and little is known regarding biofilm formation of food-related strains. We have studied biofilm formation of both food-related and clinical S. aureus strains grown under different stress conditions (temperature, sodium chloride, glucose and ethanol) relevant for food processing. Strong biofilm formers were identified among food related S. aureus strains, and biofilm formation was affected by environmental conditions relevant for the food industry. The results showed that temperatures suboptimal for growth increased the production of biofilm. The combined presence of sodium chloride and glucose enhanced the biofilm formation. Both temperature and osmolarity affected the expression of several biofilm associated genes (e.g. icaA and rbf). Variations in gene expression (e.g. icaA, agrA and sigB) between strains were also observed. Our results support the existence of both ica dependent and ica-independent mechanisms of biofilm production in S. aureus. The phenotypic and genotypic results showed highly diverse and complex patterns of biofilm formation in S. aureus. This clearly demonstrates that caution must be exercised before drawing general conclusions about gene expression in S. aureus in relation to regulation of biofilm formation. The results are relevant for food safety as they indicate that food processing conditions could promote biofilm formation by S. aureus. PMID- 17408793 TI - Germination of Bacillus cereus spores adhered to stainless steel. AB - Adhered spores of Bacillus cereus represent a significant part of the surface derived contamination in processing equipment used in the dairy industry. As germinated spores lose their resistance capacities instantaneously, efficient germination prior to a cleaning in place treatment could aid to the disinfecting effect of such a treatment. Therefore, spores of B. cereus ATCC 14579 and that of the environmental isolate B. cereus CMCC 3328 were assessed for their germination behaviour when adhered to a stainless steel surface. A mixture of l-alanine and inosine initiated germination of adhered spores efficiently, resulting in 3.2 decimal logarithms of germination. Notably, implementation of a germination inducing step prior to a representative cleaning in place procedure reduced the number of survivors with over 3 decimal log units, while an alkali treatment alone, as part of the cleaning in place procedure, did not show any effect on B. cereus spore viability. These results show that implementation of a germination step enhances the disinfection effect of currently used cleaning in place procedures. PMID- 17408794 TI - Incorporating metabolic flux ratios into constraint-based flux analysis by using artificial metabolites and converging ratio determinants. AB - One of the well-established approaches for the quantitative characterization of large-scale underdetermined metabolic network is constraint-based flux analysis, which quantifies intracellular metabolic fluxes to characterize the metabolic status. The system is typically underdetermined, and thus usually is solved by linear programming with the measured external fluxes as constraints. Thus, the intracellular flux distribution calculated may not represent the true values. (13)C-constrained flux analysis allows more accurate determination of internal fluxes, but is currently limited to relatively small metabolic networks due to the requirement of complicated mathematical formulation and limited parameters available. Here, we report a strategy of employing such partial information obtained from the (13)C-labeling experiments as additional constraints during the constraint-based flux analysis. A new methodology employing artificial metabolites and converging ratio determinants (CRDs) was developed for improving constraint-based flux analysis. The CRDs were determined based on the metabolic flux ratios obtained from (13)C-labeling experiments, and were incorporated into the mass balance equations for the artificial metabolites. These new mass balance equations were used as additional constraints during the constraint-based flux analysis with genome-scale E. coli metabolic model, which allowed more accurate determination of intracellular metabolic fluxes. PMID- 17408795 TI - Monoclonal antibody against non-dominant epitopes of HBV e Ag was raised by antigen-antibody co-immunization. AB - Detection of hepatitis B e antigen (HBeAg) in the sera of individuals infected with hepatitis B virus (HBV) can indicate both a high infectivity of the disease and a poor prognosis of disease treatment. Most of monoclonal antibodies raised against HBV e proteins interact with immuno-dominant epitopes, such as HBeAg beta. In order to raise antibodies against non-dominant epitopes of HBV e protein, in this study, mice were immunized with both recombinant HBeAg (rHBeAg) and an anti-HBeAg antibody (EWB) recognizing a dominant antigenic epitope of HBeAg (HBeAg-beta epitope). With this strategy, we successfully selected two monoclonal antibodies, S-29-3 and S-72-3. Both S-29-3 and S-72-3 bind to recombinant HBeAg with a high affinity. The epitope mapping assay determined that the S-73-2 recognizes the N-terminal of HBeAg (1-118 aa) and the S-29-3 recognizes the C-terminal of HBeAg (91-149 aa). Further experiment showed that these two antibodies could be formed a pair-Abs that is used in detecting native HBeAg from the sera of HBV patients. The conclusion is that the developed method is useful to raise mAb against non-dominant epitopes in given Ag. PMID- 17408796 TI - Performance of different small sample RNA amplification techniques for hybridization on Affymetrix GeneChips. AB - A key issue in RNA amplification techniques is the preservation of original transcript abundance, however popular high-grade RNA amplification methods lack sufficient validation regarding the potential bias of gene expression profiles. This study evaluated a double-round T7-based and a PCR-based amplification protocol, using the Affymetrix GeneChip platform. Both small sample methods performed excellently in terms of yield and reproducibility (r>0.99), and also the within-method concordance with respect to differential gene expression was as high as with standard single-round T7-based amplification. However, when comparing the overlap of all differentially expressed genes between standard and small sample methods, this was only moderate for the double-round T7 (48.7-55.0%) as well as for the PCR-based amplification protocol (51.9-58.0%). In contrast, the concordance for the top 100 genes with highest fold changes was significantly higher, indicating that both small sample methods generate reliable results when focusing on strongly regulated genes. PMID- 17408797 TI - Development of NaCl-tolerant line in Chrysanthemum morifolium Ramat. through shoot organogenesis of selected callus line. AB - Plants were regenerated successfully through shoot organogenesis of a NaCl selected callus line of Chrysanthemum morifolium Ramat. cv. Maghi Yellow (a salt sensitive cultivar), developed through stepwise increase in NaCl concentration (0 100mM) in the MS medium. The stepwise increase in NaCl concentration from a relatively low level to cytotoxic level was found to be a better way to isolate NaCl-tolerant callus line, since direct transfer of callus to high saline medium was detrimental to callus survival and growth. The selected callus line exhibited significant increase in superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) activities compared to control callus (grown in medium devoid of NaCl). Stability of salt tolerance character of the selected callus line was checked by growing the calli in NaCl free medium for 3 consecutive months followed by re-exposure to higher salinity stress (120mM NaCl). Among different growth regulator treatments, a combination of 5mgl(-1) TDZ (Thidiazuron) along with 0.25mgl(-1) NAA and 0.5mgl(-1) GA(3) was found to be the most effective for shoot organogenesis in selected callus line. The regeneration potential of the NaCl-tolerant callus ranged from 20.8% to 0% against 62.4% to 0% in control callus line. Under elevated stress condition (medium supplemented with 250mM NaCl), selected calli derived regenerants (S1 plants) exhibited significantly higher SOD and APX activities over both PC (positive control: control callus derived plants grown on MS medium devoid of NaCl) and NC (negative control: control callus derived plants subjected to 250mM NaCl stress) plants. In addition, the NC plants showed stunted growth, delayed root initiation, and had lesser number of roots as compared to S1 plants. Based on growth performance and antioxidant capacity, the S1 plants could be considered as NaCl-tolerant line showing all positive adaptive features towards the salinity stress. Further study on agronomic performance of these S1 plants under saline soil condition need to be undertaken to check the genetic stability of the induced salt-tolerance. PMID- 17408798 TI - Target-specific intracellular delivery of siRNA using degradable hyaluronic acid nanogels. AB - Novel hyaluronic acid (HA) nanogels physically encapsulating small interfering RNA (siRNA) were fabricated by an inverse water-in-oil emulsion method. Thiol conjugated HA dissolved in aqueous emulsion droplets was ultrasonically crosslinked via the formation of disulfide linkages to produce HA nanogels with a size distribution from 200 to 500 nm. Green fluorescence protein (GFP) siRNA was physically entrapped within the HA nanogels during the emulsion/crosslinking process. The HA/siRNA nanogels were readily taken up by HA receptor positive cells (HCT-116 cells) having HA-specific CD44 receptors on the surface. Release rates of siRNA from the HA nanogels could be modulated by changing the concentration of glutathione (GSH) in the buffer solution, indicating that the degradation/erosion of disulfide crosslinked HA nanogels, triggered by an intracellular reductive agent, controlled the release pattern of siRNA. When HA nanogels containing GFP siRNA were co-transfected with GFP plasmid/Lipofectamine to HCT-116 cells, a significant extent of GFP gene silencing was observed in both serum and non-serum conditions. The gene silencing effect was reduced in the presence of free HA in the transfection medium, revealing that HA nanogels were selectively taken up by HCT-116 cells via receptor mediated endocytosis. PMID- 17408799 TI - [Emotions and situations of communication in children and adolescents with frontal cerebral lesions]. AB - OBJECTIVE: Following cerebral lesions, especially right and frontal cerebral lesions, patients often have difficulties in social cognition. Emotional skills are often impaired. Adults with cerebral lesions have difficulty identifying facial emotional expressions and attributing emotions in situations of communication. However, few studies have involved children and adolescents with cerebral lesions. Our first goal was to determine whether the performance in emotional tasks of young patients with frontal cerebral lesions is as impaired as that of adults, and the second goal was to study the role of emotional cues in pragmatic aspects of language. METHOD: The study involved 7 children and adolescents with frontal cerebral lesions but no aphasia and 7 control subjects. We used two emotional tasks: one to assess the ability to identify facial emotional expressions, and one to assess the ability to attribute emotions in situations of communication. RESULTS: The control and experimental groups did not differ significantly in identifying and attributing emotions. A further analysis of responses revealed that the patients were more impaired than controls in attributing complex emotions. As well, the patients tended to produce more justifications based on irrelevant elements than the controls. CONCLUSION: Our first data have shown that the children and adolescents with frontal cerebral lesions retain the ability to analyse emotions in situations of communication but have subtle difficulty in analysing emotional cues in social situations. PMID- 17408800 TI - Strength-duration properties in diabetic polyneuropathy. AB - We sought to examine the changes in the strength-duration time constant (SDTC) of the median nerve in diabetic polyneuropathy. The SDTC is a measure of axonal excitability and depends on the biophysical properties of the axonal membrane. It may provide some information about Na(+) channel functioning. Forty-nine diabetic patients (30 men, 19 women; mean age, 54.6+/-9.4 years) and 15 age-matched healthy subjects (11 men, 4 women; mean age, 53.1+/-7.6 years) participated in the study. SDTC and rheobase values were 384.5+/-97.9 micros and 4.6+/-2.4 mA in patients and 313.8+/-46.7 micros and 6.0+/-2.6 mA in controls. The SDTC was found to be significantly different between the two groups (P=.009 for SDTC and P=.066 for rheobase). Fasting glucose and HbA1c levels of patients were 206.6+/-88.7 mg/dL and 8.8+/-2.4%, and these were not correlated with SDTC or rheobase (P<.05). We suggest that longer SDTC, indicating increased axonal excitability may develop in patients with polyneuropathy associated with uncontrolled diabetes. PMID- 17408801 TI - Role of insulin receptor substrate-4 in IGF-I-stimulated HEPG2 proliferation. AB - BACKGROUNDS/AIMS: Insulin receptor substrate-4 (IRS-4) is a scaffold protein that mediates the actions of insulin-like growth factor-I (IGF-I). Its expression increases dramatically after partial hepatectomy (a liver regeneration model). Herein, we report IRS-4 expression in a human hepatoblastoma cell line (HepG2) and IGF-I-dependent IRS-4 tyrosine phosphorylation. METHODS: The role of IRS-4 in HepG2 proliferation was established by RNA interference (siRNA). After 72h of transfection with IRS-4 siRNA, we observed a specific reduction in IRS-4 expression. RESULTS: Depletion of IRS-4 levels decreased ERK phosphorylation, p70S6K phosphorylation and IGF-I-stimulated cell proliferation. Changes in ERK phosphorylation in IRS-4-depleted cells were independent of ras/raf/MEK1/2- and PI3K/Akt-cascades. IRS-4 down-regulation abolished IGF-I-, TPA- and IGF-I plus TPA-stimulated ERK and p70S6K activities. Our results suggest that PKC-epsilon mediates the effect of IRS-4 on ERK activity. Moreover, decreased IRS-4 levels diminished FBS- and IGF-I-stimulated HepG2 growth and cause stress fiber disruption in HepG2 cell line. CONCLUSIONS: Collectively, our data suggest that IRS-4 plays an important role in HepG2 proliferation/differentiation and exerts its actions through ERK and p70S6K activation in a ras/raf/MEK1/2- and PI3Kinase/Akt-independent manner and in a PKC-dependent way. PMID- 17408802 TI - User-friendly software for the analysis of brain lesions (ABLe). AB - We previously developed a software package called ABLe (analysis of brain lesions) which characterizes brain lesions in terms of lesion volume and intersection with cytoarchitecture (e.g. Brodmann areas). Since our previous publication, there have been significant improvements to this software package which utilize methods standard to the neuroimaging community. These features include spatial normalization to the MNI template brain (standard of the international consortium for brain mapping), and use of the volume occupancy Talairach labels (VOTL) and automated anatomical labeling (AAL) atlases for full brain quantification of structures impacted by the lesion. Methods for multi subject studies including lesion-symptom mapping proposed by Bates et al. have been extended in ABLe to produce an exploratory analysis summarizing correlations between subjects with overlapping lesions and behavioral deficit. A subset of data from an ongoing traumatic head injury study correlating deficit with brain anatomy is used to demonstrate the power of this software package. PMID- 17408803 TI - Clinical policy: Critical issues in the evaluation and management of adult patients presenting to the emergency department with acute heart failure syndromes. PMID- 17408804 TI - Interoceptive "satiety" signals produced by leptin and CCK. AB - The present studies assessed the extent to which the adiposity signal leptin and the brain-gut hormone cholecystokinin (CCK), administered alone or in combination, give rise to interoceptive sensory cues like those that are produced by a low (1h) level of food deprivation. Rats were trained with cues arising from 1 to 24-h food deprivation as discriminative stimuli. For one group, 24-h food deprivation predicted the delivery of sucrose pellets, whereas 1-h food deprivation did not. Another group received the reversed deprivation level sucrose contingency. After asymptotic performance was achieved, the effects of leptin and CCK on food intake and on discrimination performance were tested under 24-h food deprivation. In Experiment 1a, leptin administered into the third cerebroventricle (i3vt) at 3.5 or 7.0 microg doses had little effect, compared to saline on food intake or discriminative responding. In Experiment 1b, leptin (7.0 microg, i3vt) combined with CCK-8 (2 microg/kg, i.p.) reduced food intake significantly, but the findings indicated that CCK-8 alone produces interoceptive discriminative cues more like those produced by 1- than 24-h food deprivation. Experiment 2a tested rats with i.p. leptin (0.3 and 0.5mg/kg). Although neither dose suppressed intake, the 0.3mg/kg dose produced interoceptive cues like 1-h food deprivation. Experiment 2b tested two doses of CCK-8 (2 and 4 mg/kg, i.p.) and found significant intake suppression and generalization of discrimination with both doses of CCK-8. These findings suggest a role for both leptin and CCK in the production of sensory consequences that correspond to "satiety". PMID- 17408805 TI - Conjugation of a novel Apaf-1 inhibitor to peptide-based cell-membrane transporters: effective methods to improve inhibition of mitochondria-mediated apoptosis. AB - We have identified a family of peptoids that inhibits in vitro the activity of the apoptosome, a macromolecular complex that activates mitochondrial-dependent apoptosis pathways. The analysis of peptide-based cell compatible delivery systems of the most active peptoid is presented. The active peptoid was then fused to cell penetrating peptides (CPP) as penetratin (PEN-peptoid) and HIV-1 TAT (TAT-peptoid). PEN-peptoid showed greater cell viability and as a consequence better efficiency as an apoptosis inhibitor than the TAT-peptoid. The intracellular trafficking of both inhibitors was studied by flow cytometry and confocal fluorescence microscopy. Finally, the influence of the cargo (peptoid) molecules on the conformational behavior of the CPP in buffers and in membrane mimetic environments was analyzed using circular dichroism (CD) spectroscopy. PMID- 17408806 TI - Lack of potentiation of bradykinin by angiotensin-(1-7) in a type 2 diabetes model: role of insulin. AB - Considering the growing importance of the interaction between components of kallikrein-kinin and renin-angiotensin systems in physiological and pathological processes, particularly in diabetes mellitus, the aim of the present study was to investigate the interaction between angiotensin-(1-7) (Ang-(1-7)) and bradykinin (BK), important components of these systems in an insulin resistance model of diabetes, and the effect of insulin on it. For this the response of mesenteric arterioles of anesthetized neonatal streptozotocin-induced (n-STZ) diabetic and control rats was evaluated using intravital microscopy. Though capable of potentiating BK in non-diabetic rats, Ang-(1-7) did not potentiate BK in n-STZ rats. Chronic but not acute insulin treatment restored the potentiation. This restorative effect of insulin was abolished by a K+ channel blocker (tetraethylammonium), by nitric oxide synthase inhibitor (N-nitro-L-arginine methyl ester) and by a cyclooxygenase inhibitor (indomethacin). On the other hand, Na(+)-,K(+)-ATPase inhibition (by ouabain) did not abolish the effect of insulin. There was no difference in mRNA and protein expression of B1 and B2 kinin receptor subtypes between n-STZ diabetic and control rats. Insulin treatment did not alter the kinin receptor expression. Our data allow us to conclude that diabetes impaired the interaction between BK and Ang-(1-7) and that insulin restores it. The restoring effect of insulin depends on membrane hyperpolarization, nitric oxide release and cyclooxygenease metabolites but not Na+K+-ATPase. Alteration of kinin receptor expression might not be involved in the restoring effect of insulin on the potentiation of BK by Ang-(1-7). PMID- 17408807 TI - Effects of ethanol on the remodeling of neutral lipids and phospholipids in brain mitochondria and microsomes. AB - We have analyzed the effects of ethanol in vitro on the remodeling of neutral lipids and phospholipids in mitochondria and microsomes isolated from chick brain. We used three different fatty acyl-CoAs of similar chain lengths but different degrees of unsaturation. Our results demonstrate the existence of active mechanisms for acyl-CoA transfer into neutral lipids and phospholipids in both mitochondria and microsomes. The profile of fatty acid incorporation was clearly different according to the membrane and lipid fraction in question. Thus, in mitochondrial lipids, the remodeling processes showed a clear preference for the saturated fatty acid whilst the polyunsaturated one was the preferred substrate for microsomal lipid acylation. With regard to the effects of ethanol in vitro, we were able to demonstrate that exposure of the membrane to ethanol led to an increase in the incorporation of polyunsaturated fatty acid into triacylglycerol (TG) in both mitochondria and microsomes, indicating that it directly stimulates the acylation of diacylglycerol (DG) to give TG. This effect may then contribute to the widely reported stimulation of TG biosynthesis in cases of both acute and chronic ethanol ingestion. It is noteworthy that the exposure of microsomes to ethanol in vitro also stimulated the incorporation of oleoyl-CoA into the aminophospholipids phosphatidylethanolamine (PE) and phosphatidylserine (PS). We also demonstrate that both mitochondria and microsomes synthesize fatty acid ethyl esters (FAEEs) from fatty acyl-CoA, although there is a clear difference in preference for the fatty acid used as substrate in the esterification of the alcohol. Thus, mitochondria were capable of forming FAEEs from the polyunsaturated fatty acid whilst in microsomes the saturated fatty acid was the preferred substrate. In both types of membrane, FAEE production was lowest with the monounsaturated fatty acyl-CoA. PMID- 17408808 TI - Psychosis of Alzheimer's disease: Gender differences in regional perfusion. AB - We sought to determine whether the presence of psychotic symptoms in patients with Alzheimer's disease is associated with abnormal regional cerebral function. Perfusion single photon emission computed tomography images from 51 AD patients with psychotic symptoms were compared to images of 52 AD patients without such symptoms. Group comparisons were made with a voxel-based method, Statistical Parametric Mapping. We found that perfusion was lower in female patients with psychotic symptoms in right infero-lateral prefrontal cortex and in inferior temporal regions compared to female patients without such symptoms. In contrast, perfusion was higher in male patients with psychotic symptoms in the right striatum compared to male patients without such symptoms. Comparison groups did not differ in age or in dementia severity, as estimated by the Mini-Mental State Examination (MMSE). These results support the role of right hemisphere prefrontal and lateral temporal cortex in the psychosis of AD in women but not in men, and raise the possibility that these dysfunctional processes have a gender-specific regional pathophysiology in AD. PMID- 17408809 TI - Evaluation of the anti-inflammatory and anti-nociceptive activities of novel synthesized melatonin analogues. AB - This study aimed at evaluation of the reactivity of melatonin (1) towards various chemical reagents to produce new melatonin analogues containing heterocyclic moieties which would provide basic pharmacological activities. The pyrrolo[1,2 a]indole derivatives 3, 5, 12, 14 and pyrido[1,2-a]indole derivatives 9a, b were synthesized via straightforward and efficient methods and their structures were established based on the analytical and spectral data. Also, this work was extended to study the potential role of the novel synthesized melatonin analogues 3, 5, 9a and 12 as anti-inflammatory and anti-nociceptive agents in comparison with melatonin. After s.c. administration all compounds induced significant anti inflammatory activity, inhibiting the paw oedema response compared with the control group at all time points in the test. Compound 5 has the strongest anti inflammatory activity which exceeds that of the parent reference, melatonin, followed by compounds 9a and 12, at the first 2h of administration. Effect of melatonin analogues on thermal pain, acetic acid-induced writhing and gastric lesions caused by indomethacin was also investigated. Compounds 5 and 12 were more potent as anti-nociceptive drugs; they are more potent in this respect than melatonin. PMID- 17408810 TI - QSAR analysis of 1,4-dihydro-4-oxo-1-(2-thiazolyl)-1,8-naphthyridines with anticancer activity. AB - In the present study a quantitative structure activity relationship (QSAR) analysis was applied to a series of 100 of 7- and 3-substituted 1,4-dihydro-4-oxo 1-(2-thiazolyl)-1,8-naphthyridine derivatives. The Chem-X (version 2000) software was used to develop 3D QSAR models. The steric and electrostatic interactions between a probe atom (H(+)) and a set of aligned molecules were assessed using the comparative molecular field analysis method. Statistically relevant models were derived for both electrostatic and steric fields. A 2D model over a restricted series of close structural analogs was derived as well. A number of conclusions on the relationship between the type and size of different substituents and the antitumor activity of the compounds were derived. PMID- 17408811 TI - N- and S-alpha-l-arabinopyranosyl[1,2,4]triazolo[3,4-b][1,3,4]thiadiazoles. First synthesis and biological evaluation. AB - First synthesis of N- and S-alpha-l-arabinopyranosyl[1,2,4]triazolo[3,4 b][1,3,4]thiadiazoles is described. Antimicrobial screening of two selected regioisomeric compounds against Aspergillus fumigatus, Penicillium italicum, Syncephalastrum racemosum, Candida albicans, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis and Escherichia coli are compared. PMID- 17408812 TI - Spectroscopic studies of DNA binding modes of cation-substituted anthrapyrazoles derived from emodin. AB - The DNA binding properties of three cation-substituted anthrapyrazole derivatives of emodin with calf thymus DNA were characterized by spectroscopic methods and the specific binding modes were elucidated. At low drug and high DNA concentrations, compound 1 with a mono-cationic amino side chain exhibited an intercalative binding mode, 2 with a much longer and more flexible di-cationic side chain exhibited an external binding mode, and 3 with a rigid di-cationic side chain exhibited both intercalative and external binding modes. The DNA binding mode of compounds was altered after structural modification. The molecular structure-DNA binding relationships found from this study may be useful for the design of anthrapyrazole derivatives with desired binding characteristics. PMID- 17408813 TI - Using the critical incident technique to explore student nurses' perceptions of language awareness. AB - Current evidence demonstrates that language awareness in nursing contributes to high quality patient care. Although language awareness amongst qualified nurses has been considered, there is a dearth of literature that examines how student nurses respond to language sensitivity in the healthcare setting. The critical incident technique was used to elicit the perceptions of student nurses regarding language awareness. Twenty students produced written descriptions of incidents that they had encountered during clinical practice and they subsequently reflected on these incidents. Data were analysed using framework analysis whereby meaning units were organised into themes and categories. Four distinct themes emerged from the analysis; namely, facilitating language choice, vulnerable groups, quality of care and student learning. Each theme was characterised by a number of subcategories. Student nurses identified positive and negative instances of language matters in health care that are driven by personal and organisational strategies and impact on patient care and student learning. Student nurses were conscious of the influence of language awareness on patient care and noted that structural and process elements relating to language awareness were in play in the healthcare setting. Moreover, they indicated that the consequent outcomes of these elements influenced the quality of care and student learning. PMID- 17408814 TI - Managerial perceptions of mentor, lecturer practitioner and link tutor roles. AB - Educating pre-registration nurses in clinical practice is a global issue. Within different countries problems exist in educating and supervising students in clinical practice and various models of clinical education are employed. In Wales, United Kingdom, this responsibility is divided between mentors, lecturer practitioners and link tutors. This paper reports on the third phase of a three phase study in Wales to explore differences between mentors, lecturer practitioners and link tutors, and how they work together to assist students to integrate theory and practice. Four focus group interviews of National Health Service managers and Higher Education managers (n=22) were conducted. Qualitative content analysis revealed four themes: role characteristics and competencies, role differences, role conflict, and future options. The findings suggest a theory-practice continuum along which mentors, lecturer practitioners and link tutors occupy different positions. The article explores these different positions and offers suggestions for future role development. PMID- 17408815 TI - A single DNA immunization in combination with electroporation prolongs the primary immune response and maintains immune memory for six months. AB - Recombinant protein vaccines and vaccines using killed or inactivated pathogens frequently require multiple vaccinations to induce protective immune responses which may be of relatively short duration. Furthermore, increasing concern regarding adverse local and systemic reactions to injected vaccines is driving the quest for vaccine formulations, which induce protective immunity following a single administration. Vaccine studies frequently evaluate immune responses and disease protection within a relatively short interval following primary and secondary immunizations and, therefore, fail to address the duration of immunological memory or protection. DNA vaccines offer a unique opportunity to enhance the duration of immune responses through their capacity for prolonged antigen expression. The route of DNA vaccination and the method of plasmid delivery are critical factors, which can determine transfection efficiency and the duration of vaccine antigen production. Studies were completed which demonstrated that a single intramuscular DNA vaccination, when combined with electroporation, significantly enhanced the onset and duration but not the magnitude of the primary antibody response. A secondary protein vaccination was performed 6 months after the primary DNA immunization. A significant (p < or = 0.0001) correlation was observed between both the magnitude (r2 = 0.40) and duration (r2 = 0.74) of the primary antibody response and the occurrence of a secondary antibody response. Therefore, an effective primary DNA vaccination has the potential to significantly prolong the duration of an antibody response and possibly reduce the frequency of revaccination. PMID- 17408816 TI - Unexplained sudden death, including sudden infant death syndrome (SIDS), in the first and second years of life: case definition and guidelines for collection, analysis, and presentation of immunization safety data. PMID- 17408817 TI - Long-term colonisation-inhibition studies to protect broilers against colonisation with Salmonella Enteritidis, using Salmonella Pathogenicity Island 1 and 2 mutants. AB - Mutants in the Salmonella Pathogenicity Island 1 (hilA and sipA) and 2 (ssrA) were tested for their potential to induce protection against infection by homologous virulent Salmonella Enteritidis challenge strain, administered 24h later, in chickens. Although they colonised the internal organs to a significantly lower degree compared to the wild type strain, both a sipA and a ssrA mutant persistently colonised the gut when inoculated to newly hatched chicks. After inoculation of 1-day-old chicks with a sipA or a ssrA mutant and subsequent challenge with a wild type Salmonella Enteritidis 24h later, a significant degree of resistance against caecal and internal organ colonisation by the challenge strain was found. The protection lasted for the full 6 weeks of study, but due to their persistence, the sipA and ssrA mutants are not useful to induce broiler protection. After inoculation of newly hatched chicks with a hilA mutant no positive cloacal swabs could be detected anymore at 4 weeks post inoculation and the hilA mutant was almost completely cleared from the gut. When newly hatched chicks were inoculated with a hilA mutant and challenged 24h later, the excretion of the virulent challenge strain was significantly reduced and the intestinal colonisation of the challenge strain was inhibited to a high level until the age of 9 days. Moreover, the hilA mutant exerted a significant profound inhibition of internal organ colonisation by the virulent challenge strain throughout the study period. The approach of vaccination with a hilA mutant strain can be a valuable basis for development of vaccine strains for broilers protection. PMID- 17408818 TI - Aerosol and subcutaneous measles vaccine: measles antibody responses 6 years after re-vaccination. AB - There are no large-scale data on the long-term persistence of measles antibody after vaccination by the aerosol route. We therefore followed-up South African schoolchildren 6 years after their re-vaccination with Edmonston-Zagreb (EZ) and Schwarz (SW) measles vaccine given by aerosol and subcutaneous routes. Measles antibody levels and the proportion of children who were seropositive at year 6 remained significantly higher in the Edmonston-Zagreb aerosol group compared to the groups that received Schwarz or Edmonston-Zagreb vaccine subcutaneously. Measles re-vaccination by aerosol evokes a stronger and much longer lasting antibody response than injected vaccine and should thus provide more durable protection against measles. PMID- 17408819 TI - Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity. AB - Ricin is a potent toxin associated with bioterrorism for which no vaccine or specific countermeasures are currently available. A stable, non-toxic and immunogenic recombinant ricin A-chain vaccine (RTA 1-33/44-198) has been developed by protein engineering. We identified optimal formulation conditions for this vaccine under which it remained stable and potent in storage for up to 18 months, and resisted multiple rounds of freeze-thawing without stabilizing co solvents. Reformulation from phosphate buffer to succinate buffer increased adherence of the protein to aluminum hydroxide adjuvant from 15 to 91%, with a concomitant increase of nearly threefold in effective antigenicity in a mouse model. Using Fourier-transform infrared spectroscopy, we examined the secondary structure of the protein while it was adhered to aluminum hydroxide. Adjuvant adsorption produced only a small apparent change in secondary structure, while significantly stabilizing the protein to thermal denaturation. The vaccine therefore may be safely stored in the presence of adjuvant. Our results suggest that optimization of adherence of a protein antigen to aluminum adjuvant can be a useful route to increasing both stability and effectiveness, and support a role for a "depot effect" of adjuvant. PMID- 17408820 TI - Indoor radon monitoring in Northern Iran using passive and active measurements. AB - In this work we present the results of a 2-year survey of indoor radon variations in four cities of Lahijan, Ardabil, Sar-Ein and Namin in North and Northwest Iran. We used both passive and active measurements by solid state nuclear track detectors (SSNTDs) with CR-39 polycarbonate and PRASSI Portable radon Gas Surveyor. A total of 1124 samplers in Lahijan, Ardabil, Sar-Ein and Namin were installed. Sampling frequency was seasonal and sampling locations were randomly chosen based on dwelling structures, floors, geological formations, elevation and temperature variation parameters. For quality assurance, 281 active measurements and double sampling were carried out. Based on our results and the results of previous surveys, Ardabil and Lahijan have the second and third highest radon concentration in Iran, respectively (Ramsar is first). The average radon concentration during the year in Lahijan, Ardabil, Sar-Ein and Namin were 163, 240, 160 and 144 Bq/m(3) with medians of 160, 168, 124 and 133 Bq/m(3), respectively. These concentrations give rise to annual effective doses of 3.43 mSv/y for Lahijan and 5.00 mSv/y for Ardabil. The maximum recorded concentration was 2386 Bq/m(3) during winter in Ardabil and the minimum concentration was 55 Bq/m(3) during spring in Lahijan. Relationships between radon concentration and building materials and room ventilation were also studied. The dosimetry calculations showed that these four cities could be categorized as average natural radiation zones. The correlation coefficients relating warm and cold season radon variation data were obtained. PMID- 17408821 TI - Antenatal taboos among Chinese women in Hong Kong. AB - OBJECTIVE: to identify the antenatal taboos commonly practised by pregnant Hong Kong Chinese women; to explore the health beliefs behind these taboos; and to examine how pregnant women perceived and reacted to the cultural tradition. DESIGN: general ethnography and in-depth interviews, followed by a quantitative self-reported survey. SETTING: Antenatal clinic of a university-affiliated hospital in Hong Kong. PARTICIPANTS: consecutive samples of 60 women for in-depth interviews, and 832 women for the survey. MEASUREMENTS: an inventory on the adherence and attitude towards antenatal taboos, and the Beck Depression Inventory that measures severity of depression. FINDINGS: antenatal taboos were still commonly observed by contemporary Chinese women. Miscarriage, fetus malformation and fetal ill-health were the key cultural fears that drove contemporary Chinese women to observe the traditional taboos. About one-quarter and one-tenth of the women, respectively, felt unhappy and disputed with their families about the taboos. These women had significantly higher levels of depression in late pregnancy and during childbirth. IMPLICATIONS FOR PRACTICE: health-care practitioners should be aware of the benefits and risks of traditional antenatal taboos on maternal health. Although some taboos can be socio-morally protective, the tension created by the observation of cultural tradition in modernity may impair maternal psychological well-being. Health-care providers in Western countries should be vigilant of the complex cultural tension faced by migrant Chinese mothers. PMID- 17408822 TI - A descriptive study of the use of the Internet by women seeking pregnancy-related information. AB - OBJECTIVE: to investigate whether pregnant Swedish women used the Internet to retrieve pregnancy-related information, how they perceived the reliability of the information, and whether they discussed this information with their midwife during antenatal care visits. DESIGN AND SETTING: a descriptive, cross-sectional design using waiting-room questionnaires to obtain information from pregnant women attending 11 antenatal clinics in a county in mid-Sweden during 2004. PARTICIPANTS: all Swedish-speaking women who visited the clinics during a 2-week period, and who were at least 32 weeks pregnant, were invited to participate. A total of 182 women, with a mean age of 31 years, participated in the study; the response rate 85%. FINDINGS: most (91%) of the women had access to the Internet and, to a great extent (84%), used it to retrieve information, most often in the early stages of their pregnancy. Fetal development and stages of childbirth were the two most often mentioned topics of interest. Most participants considered the information to be reliable, and the two most important criteria for judging the trustworthiness of web-based information were if the facts were consistent with information from other sources and if references were provided. Most (70%) of the women did not discuss the information they had retrieved from the Internet with their midwife, but more than half of them (55%) searched for information on topics brought up by the midwife. CONCLUSION: Swedish pregnant women often use the Internet to find information on various topics related to pregnancy, childbirth and the expected baby. They perceive the information to be reliable but rarely discuss it with their midwives. Antenatal care providers should be able to guide pregnant women to high-quality, web-based information and then take the opportunity to discuss this information with them during antenatal visits. PMID- 17408823 TI - Effect of chemophytostabilization practices on arbuscular mycorrhiza colonization of Deschampsia cespitosa ecotype Warynski at different soil depths. AB - The effects of chemophytostabilization practices on arbuscular mycorrhiza (AM) of Deschampsia cespitosa roots at different depths in soils highly contaminated with heavy metals were studied in field trials. Mycorrhizal parameters, including frequency of mycorrhization, intensity of root cortex colonization and arbuscule abundance were studied. Correlations between concentration of bioavailable Cd, Zn, Pb and Cu in soil and mycorrhizal parameters were estimated. An increase in AM colonization with increasing soil depth was observed in soils with spontaneously growing D. cespitosa. A positive effect of chemophytostabilization amendments (calcium phosphate, lignite) on AM colonization was found in the soil layers to which the amendments were applied. Negative correlation coefficients between mycorrhizal parameters and concentration of bioavailable Cd and Zn in soil were obtained. Our results demonstrated that chemophytostabilization practices enhance AM colonization in D. cespitosa roots, even in soils fertilized with high rates of phosphorus. PMID- 17408824 TI - Geochemical and isotopic constraints on groundwater-surface water interactions in a highly anthropized site. The Wolfen/Bitterfeld megasite (Mulde subcatchment, Germany). AB - The Bitterfeld/Wolfen region is a megasite with multiple contaminant sources from more than a century of industrial activity, which have a considerable impact on the environment. At present, the contaminated groundwater covers an area of about 25km(2) and poses a threat for the surrounding aquifers and the Mulde River. This study focuses on the Schachtgraben, a man-made channel in the Mulde Floodplain that collects the effluents of the industrial area. It aims to characterise the relationship between surface water (channels, rivers) and the groundwater in the shallow Quaternary aquifer. Waters are Ca-SO(4) type with TDS reaching 3.8gL(-1) in the industrial area. Stable isotopes (delta(18)O, delta(2)H) show that two of the rivers are recharged mainly by groundwater that can be divided into two groups. Strontium isotopes ((87)Sr/(86)Sr) designate different geochemical end members and enable the identification of mixing between natural and anthropogenic surface and groundwater. PMID- 17408825 TI - Evaluation of the health impact of lowering the formaldehyde occupational exposure limit for Quebec workers. AB - This study aimed at assessing the impact on irritating effects of lowering the current occupational exposure limit (OEL) for formaldehyde in occupational settings in the Province of Quebec, Canada, from a 2 ppm ceiling value to 1, 0.75 or 0.3 ppm. This was achieved through (i) a re-assessment of the exposure response relationship based on a pooled analysis of published controlled human studies on the incidence of the most sensitive effects related to acute formaldehyde exposure (irritation of the eyes, nose and throat) and (ii) application of this relationship to the data on current exposure to formaldehyde in industrial sectors of Quebec. Results show that the exposure-irritating effect relationship compiled by concentration ranges and by degree of severity was best described by quadratic regression. Considering the current distribution of formaldehyde exposure among the 143,491 Quebec workers concerned, eye irritation, the most sensitive effect, could affect 526 workers (0.367%) at a moderate degree and 50 workers (0.035%) at a severe degree. By reducing the OEL to 1, 0.75 and 0.3 ppm, the proportion of these effects estimated to be avoided would be 442/526 (84%), 526/526 (100%) and 526/526 (100%), respectively. Results for nose and throat irritation follow the same trend. The greatest gain would thus be obtained by respecting the current OEL; the additional gain was estimated to be negligible below 0.75 ppm. The level of 0.75 ppm can be considered as a safe level that allows protecting virtually all workers. PMID- 17408826 TI - Scaling-up a public health innovation: a comparative study of post-abortion care in Bolivia and Mexico. AB - Post-abortion care (PAC), an innovation for treating women with complications of unsafe abortion, has been introduced in public health systems around the world since the 1994 International Conference on Population and Development (ICPD). This article analyzes the process of scaling-up two of the three key elements of the original PAC model: providing prompt clinical treatment to women with abortion complications and offering post-abortion contraceptive counseling and methods in Bolivia and Mexico. The conceptual framework developed from this comparative analysis includes the environmental context for PAC scale-up; the major influences on start-up, expansion, and institutionalization of PAC; and the health, financial, and social impacts of institutionalization. Start-up in both Bolivia and Mexico was facilitated by innovative leaders or catalyzers who were committed to introducing PAC services into public health care settings, collaboration between international organizations and public health institutions, and financial resources. Important processes for successful PAC expansion included strengthening political commitment to PAC services through research, advocacy, and partnerships; improving health system capacity through training, supervision, and development of service guidelines; and facilitating health system access to essential technologies. Institutionalization of PAC has been more successful in Bolivia than Mexico, as measured by a series of proposed indicators. The positive health and financial impacts of PAC institutionalization have been partially measured in Bolivia and Mexico. Other hypotheses--that scaling-up PAC will significantly reduce maternal mortality and morbidity, decrease abortion-related stigma, and prepare the way for efforts to reform restrictive abortion laws and policies--have yet to be tested. PMID- 17408827 TI - Psychosocial resources and depression among chronically ill young adults: are males more vulnerable? AB - This population-based study examined the association between chronic illness and depression and the role of psychosocial resources (coping styles, locus of control (LOC) and social support) in this association, among young Finnish adults aged 32. Gender differences in these phenomena were also investigated. The study was based on questionnaire data from a Finnish cohort study. Participants with self-reported chronic illness (e.g. diabetes, asthma, migraine) were grouped together (n=257) and compared to healthy controls (n=664). The results showed that the chronically ill males were more depressed than healthy control males. They also used more emotion-focused coping, had a more external LOC and were less often married or cohabiting than healthy males. The association between chronic illness and depression among males attenuated when the effects of emotion-focused coping disposition and LOC were taken into account, indicating a possible mediating role for these resources. Among females no differences were found in depression or psychosocial resources between the chronically ill and healthy control groups. Psychosocial resources, especially LOC, explained the gender difference in the association between chronic illness and depression. Only a few buffering effects of psychosocial resources emerged: an active problem-solving coping disposition among the chronically ill males and perceived social support among the chronically ill females seemed to act as buffers against depression. The results indicated a significant gender disparity in the association between chronic illness and depression among young adults and emphasised the role of psychosocial resources in this context. With regard to prevention we suggest that, chronically ill young adult males should be recognised as a risk group for depression that would probably benefit from guidance in learning more active coping skills and maintaining a sense of personal control in facing chronic physical illness. PMID- 17408828 TI - Pointing with the left and right hands in congenitally blind children. AB - Congenitally blind and blindfolded sighted children at ages of 6, 8, 10 and 12 years performed a pointing task with their left and right index fingers at an array of three targets on a touch screen to immediate (0 s) and delayed (4 s) instructions. Accuracy was greater for immediate than delayed pointing and there was an effect of delay for the orientation of the main axis of the pointing distribution in both groups, indicating distinct spatial representations with development such as ego- and allocentric frames of reference, respectively. The pointing responses of the blind covered less surface area indicating better overall accuracy as compared to the sighted blindfolded. The hands differed for four of the six precision and accuracy parameters. The right hand performed better and seemed relatively contextually oriented, whereas the responses of the left hand were closer to the body and egocentrically oriented. The elongation of the scatter of the pointing responses was greater for the boys and more allocentrically oriented, indicating gender differences in spatial representation. The study provides a first evidence of ego- and allocentric spatial frames of reference in congenitally blind children and an ability to point at targets with the left and right hands in the total absence of vision. PMID- 17408829 TI - Dissociation of short- and long-term face memory: evidence from long-term recency effects in temporal lobe epilepsy. AB - We tested whether memory deficits in temporal lobe epilepsy (TLE) are better described by a single- or dual-store memory model. To this aim, we analyzed the influence of TLE and proactive interference (PI) on immediate and 24-h long-term recency effects during face recognition in 16 healthy participants and 18 right and 21 left non-surgical TLE patients. PI in healthy participants or TLE erased the long-term recency effect, but left the immediate recency effect unaffected. Although the immediate recency effect was still visible in right TLE patients, the number of detected recency items during immediate recognition was decreased in right TLE compared to left TLE. Right TLE was also related to decreased detection of pre-recency items during delayed recognition compared to left TLE, and decreased detection of pre-recency items during immediate recognition under PI. The results show that the temporal lobes are necessary for the long-term recency effect, but not for the immediate recency effect, and thus speak for a dissociation of short- and long-term memory for faces. Right TLE is related to more severe long-term memory deficits than left TLE and is also related to additional short-term memory deficits for faces. PMID- 17408830 TI - No association between the neuronal pentraxin II gene polymorphism and autism. AB - Autism (MIM 209850) is a neurodevelopmental disorder characterized by difficulties with verbal and non-verbal communication, impairments in reciprocal social interactions, and displays of stereotypic behaviors, interests and activities. Twin and family studies have indicated a robust role of genetic factors in the development of autism. Neuronal Pentraxin II (NPTX2) is located in chromosome 7q21.3-q22.1, where it is a candidate region for autism. NPTX2 promotes neuritic outgrowth and is suggested to mediate uptake of degraded synaptic material during synapse formation and remodeling. NPTX2 is also associated with the clustering of synaptic AMPA receptors. It was reported that glutamate systems including AMPA receptor was associated to the pathophysiology of autism. Thus, the NPTX2 gene is involved in neuritic outgrowth, synapse remodeling and the aggregation of neurotransmitter receptors at synapses. These functions play an important role in the mechanisms of learning and brain development. In the present study, we tested for the presence of the association of four single nucleotide polymorphisms (SNPs) of NPTX2 and haplotypes consisting of the SNPs with autism, between autistic patients (n=170) and normal controls (n=214) in a Japanese population. No significant difference was observed in the allele, genotype or haplotype frequencies between the patients and controls. Thus, the NPTX2 locus is not likely to play a major role in the development of autism. However, further studies with larger sample size and sequencing of NPTX2 gene are needed to exclude a role of NPTX2 gene in autism. PMID- 17408831 TI - Sugars, carbonyls and smoke. PMID- 17408832 TI - The bioactivity of processed garlic (Allium sativum L.) as shown in vitro and in vivo studies on rats. AB - Garlic (Allium sativum L.) is widely used as an obligatory part in many cooked dishes loosing during this process a certain part of its bioactivity. Antioxidant capacity measured by the ferric-reducing/antioxidant power (FRAP) method and by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical assay was the highest in raw and in a short time processed garlic samples by cooking. 70 Wistar rats were randomly divided into 10 diet groups, each of seven. They were named Control, NPG, PG1, PG2, PG3, Chol, Chol/NPG, Chol/PG1, Chol/PG2 and Chol/PG3. The rats of the Control group were fed basal diet (BD), which included wheat starch, casein, soybean oil, cellulose, mineral and vitamin mixtures. To the BD of the nine other groups were added 25mg of lyophilized non processed garlic equivalent of 500 mg non processed garlic/kg body weight (NPG), the same quantity of processed garlic for 20, 40 and 60 min for PG1, PG2 and PG3, respectively, 1% of cholesterol (Chol), 1% of cholesterol and 25mg/kg body weight of lyophilized non processed garlic (Chol/NPG), 1% of cholesterol and the same quantity of processed garlic for 20, 40 and 60 min for Chol/PG1, Chol/PG2 and Chol/PG3, respectively. The dose of 500 mg (25mg of lyophilized garlic/kg body weight) was chosen as the most effective (Banerjee, S.K., Maulik, M., Mancahanda, S.C., Dinda, A.K., Gupta, S.K., Maulik, S.K., 2002. Dose-dependent induction of endogenous antioxidants in rat heart by chronic administration of garlic. Life Sciences 70, pp. 1509-1518). Plasma lipid profile and the total antioxidant capacity in rats significantly differed in diet groups with addition of garlic samples cooked for a long time. In summary, garlic cooked for a short time preserves a high bioactivity of non processed garlic. The diet supplemented with these samples and cholesterol improved lipid indices, decreased fibrinogen and increased antioxidant activity in plasma of rats. Therefore, for preservation of garlic bioactivity optimal regime has to be used. PMID- 17408833 TI - A comparison of chemical, antioxidant and antimicrobial studies of cinnamon leaf and bark volatile oils, oleoresins and their constituents. AB - The antioxidant, antifungal and antibacterial potentials of volatile oils and oleoresin of Cinnamomum zeylanicum Blume (leaf and bark) were investigated in the present study. The oleoresins have shown excellent activity for the inhibition of primary and secondary oxidation products in mustard oil added at the concentration of 0.02% which were evaluated using peroxide, thiobarbituric acid, p-anisidine and carbonyl values. Moreover, it was further supported by other complementary antioxidant assays such as ferric thiocyanate method in linoleic acid system, reducing power, chelating and scavenging effects on 1,1'-diphenyl-2 picrylhydrazyl (DPPH) and hydroxyl radicals. In antimicrobial investigations, using inverted petriplate and food poison techniques, the leaf and bark volatile oils has been found to be highly effective against all the tested fungi except Aspergillus ochraceus. However, leaf oleoresin has shown inhibition only for Penicillium citrinum whereas bark oleoresin has caused complete mycelial zone inhibition for Aspergillus flavus and A. ochraceus along with Aspergillus niger, Aspergillus terreus, P. citrinum and Penicillium viridicatum at 6 microL. Using agar well diffusion method, leaf volatile oil and oleoresin have shown better results in comparison with bark volatile oil, oleoresin and commercial bactericide, i.e., ampicillin. Gas chromatographic-mass spectroscopy studies on leaf volatile oil and oleoresin resulted in the identification of 19 and 25 components, which accounts for the 99.4% and 97.1%, respectively of the total amount and the major component was eugenol with 87.3% and 87.2%, respectively. The analysis of cinnamon bark volatile oil showed the presence of 13 components accounting for 100% of the total amount. (E)-cinnamaldehyde was found as the major component along with delta-cadinene (0.9%), whereas its bark oleoresin showed the presence of 17 components accounting for 92.3% of the total amount. The major components were (E)-cinnamaldehyde (49.9%), along with several other components. PMID- 17408834 TI - Evaluation of estrogenic activity of organic biocides using ER-binding and YES assay. AB - Biocides have been used not only in everyday items such as clothes, kitchenware, daily necessities, and infant utensils, but also in cosmetics and wrapping papers for foodstuffs. Since there is a high possibility of exposure to biocides from such materials, their safety must be assessed adequately using a range of methods. We investigated the estrogenic activity of 20 organic biocides using two in vitro screen assays: estrogen receptor (ER) binding assay and yeast estrogen screen (YES) assay. Twelve of the biocides were positive in the ER-binding assay. Regardless of the presence or absence of rat S9Mix for metabolic activation, 4 chloro-3-methylphenol was positive in both ER-binding and YES assay. 4-Chloro-3,5 dimethylphenol was positive in the ER-binding assay and showed a pseudopositive manner in the YES assay that was observed the dose-independent estrogenic activity at only one dose point. Hiba oil was positive in the ER-binding assay but was positive in the YES assay only in the presence of rat S9Mix. These results suggest that ER-binding and YES assay could be adapted for evaluation of the endocrine-disrupting activity of biocides. The biocides found to be positive in vitro now require assessment by in vivo screening methods. PMID- 17408835 TI - Evaluation of ambrein and epicoprostanol for their antioxidant properties: protection against adriamycin-induced free radical toxicity. AB - Ambrein and epicoprostanol were evaluated for their antioxidant potential in vitro by chemiluminescence (CL), as well as in vivo using lipid peroxides and glutathione levels as indicators in liver tissue of rats treated with adriamycin (doxorubicin) a well known free radicals producing drug. In the in vitro test, the inhibition in CL by ambrein was dose dependent. Both the high concentrations of ambrein (20-40 microg/ml) inhibited CL response significantly (P<0.05 and P<0.01, respectively) when compared to control. Similarly two low concentrations (5-20 microg/ml) of epicoprostanol inhibited CL significantly (P<0.001 and P<0.01, respectively) in comparison of DMSO control. The high concentration (40 microg/ml) of epicoprostanol behaved exceptionally and caused an increase in CL response that was more than control and significantly (P<0.001) higher than both the low concentrations. In the in vivo studies adriamycin treatment significantly (P<0.05) increased malondialdehyde (MDA) and decreased non-protein sulfhydryl (NP SH) contents in the liver tissue of mice after 5 days treatment. Ambrein (25 and 50 mg/kg) treatment as a solo therapy at both the dose levels significantly (P<0.001) decreased MDA contents in the liver tissue. On the other hand, in the combined treatment the high dose effectively prevented any rise in MDA contents and it remained around the levels of ambrein alone. In the same experiment, adriamycin declined NP-SH contents significantly (P<0.001). Ambrein alone at both the dose levels caused a decline (P<0.01) in NP-SH contents when compared to adriamycin group. But in the combined treatment this decline in NP-SH was significantly (P<0.05) different from adriamycin alone. In the experiments dealing with epicoprostanol, adriamycin treatment increased MDA contents significantly (P<0.05) that declined significantly (P<0.001) with epicoprostanol (10- or 20mg/kg) treatment. In the same experiment co-treatment with adriamycin prevented any rise in MDA contents significantly (P<0.001) as it was observed in adriamycin alone group. Although, this treatment failed to prevent any decline in NP-SH contents either alone or in combination with adriamycin. Epicoprostanol itself had the comparative declining effect on the contents of NP-SH as seen in adriamycin group. From the results of our experiments it seems that ambrein at all concentrations behaves like antioxidant in in vitro studies but the same time it decreased NP-SH contents in vivo accompanied by a decline in MDA contents. Whereas, epicoprostanol at two low concentrations had a decline in CL indicating a possible antioxidant potential but the high concentration increased CL showing a tendency towards oxidant prospective. However, in animal studies it has shown a clear protection against adriamycin induced free radical damage. PMID- 17408836 TI - [Reconstruction of hard palate defects using a facial artery musculomucosal flap]. AB - The facial artery musculomucosal flap (FAMM) is usually used in reconstruction of defects in oral cavity, most frequent being the mouth floor. This flap has many advantages from its surgical procedure, its great reliability, its mucous tissue origin, its usable surface, its large axis of rotation and the little of after effects of the donor site. The hard palate defects with bucconasal communication can raise difficulties of reconstruction. We show, by a case report, the many advantages of FAMM flap in these reconstructions. PMID- 17408837 TI - [Chromic acid burns: systematic prevention of systemic toxicity]. AB - Chromic acid burns can lead to systemic toxicity by cutaneous absorption of the chrome seen surfaces more than 1% of the total body surface area. In order to illustrate the necessity of anticipate systematically this toxicity by a specific treatment, we describe the case of a patient with systemic toxicity in the least severe situation of chromic acid burn: the chromic acid was diluted to 0,02%, the burn was superficial second degree, both thermic and chemical, on the forearm, and extended only to 1% of the total body surface area. In spite of the specific treatment, our patient had a blood transfer of the chrome, however without any consequences on the renal and hepatic functions. He cicatrised in 2 weeks, and his blood and urinary chromium levels were normalised in 3 weeks. Without this specific early treatment, what would have been the consequences of a systemic toxicity even more important? PMID- 17408838 TI - [Tissular expansion and tuberous breasts: 10 cases report]. AB - INTRODUCTION: Tuberous breast is a deficiency of mammary development, principally in the lower lateral and medial quadrants. In types II and III of the Grolleau classification, we perform a two-stage correction by tissue expansion associated with an implant in order to avoid the double bubble deformity and the difficulty of medial infra-mammary fold reconstruction. MATERIALS AND METHODS: Between February 2003 and February 2006, ten patients have been treated by implantation of an anatomical expander associated with a vertical and horizontal Puckett plasty. The second operation consisted in the replacement of the expander by an anatomical prosthesis associated, if necessary, with a second Puckett plasty and lipofilling. RESULTS: There was no double bubble deformity. Three patients required a complementary lipofilling to improve the result of the lower medial fold. One prosthesis had to be repositioned and one expander infection was responsible of a three-month delay in the reconstruction procedure. CONCLUSION: Tuberous breast correction is a therapeutic challenge in quite psychologically distressed patients. In types II and III, tissue expansion associated with a Puckett plasty increase the volume of the lower pole and reduces the risk of the double bubble deformity. The implantation of an anatomical prosthesis gives more natural effect to the contour of the glandular lower pole. This therapeutic attitude gives the chance of achieving a completely satisfactory result. PMID- 17408839 TI - Protective effects of S+ ketamine and atropine against lethality and brain damage during soman-induced status epilepticus in guinea-pigs. AB - Soman poisoning is known to induce full-blown tonic-clonic seizures, status epilepticus (SE), seizure-related brain damage (SRBD) and lethality. Previous studies in guinea-pigs have shown that racemic ketamine (KET), with atropine sulfate (AS), is very effective in preventing death, stopping seizures and protecting sensitive brain areas when given up to 1h after a supra-lethal challenge of soman. The active ketamine isomer, S(+) ketamine (S-KET), is more potent than the racemic mixture and it also induces less side-effects. To confirm the efficacy of KET and to evaluate the potential of S-KET for delayed medical treatment of soman-induced SE, we studied different S-KET dose regimens using the same paradigm used with KET. Guinea-pigs received pyridostigmine (26 microg/kg, IM) 30min before soman (62 microg/kg, 2 LD(50), IM), followed by therapy consisting of atropine methyl nitrate (AMN) (4 mg/kg, IM) 1min following soman exposure. S-KET, with AS (10mg/kg), was then administered IM at different times after the onset of seizures, starting at 1h post-soman exposure. The protective efficacy of S-KET proved to be comparable to KET against lethality and SRBD, but at doses two to three times lower. As with KET, delaying treatment by 2h post poisoning greatly reduced efficacy. Conditions that may have led to an increased S-KET brain concentration (increased doses or number of injections, adjunct treatment with the oxime HI-6) did not prove to be beneficial. In summary, these observations confirm that ketamine, either racemic or S-KET, in association with AS and possibly other drugs, could be highly effective in the delayed treatment of severe soman intoxication. PMID- 17408840 TI - The role of thiols, dithiols, nutritional factors and interacting ligands in the toxicology of mercury. AB - Mercury has been a known as a toxic substance for centuries. Whilst the clinical features of acute mercury poisoning have been well described, chronic low dose exposure to mercury remains poorly characterised and its potential role in various chronic disease states remains controversial. Low molecular weight thiols, i.e. sulfhydryl containing molecules such as cysteine, are emerging as important factors in the transport and distribution of mercury throughout the body due to the phenomenon of "Molecular Mimicry" and its role in the molecular transport of mercury. Chelation agents such as the dithiols sodium 2,3 dimercaptopropanesulfate (DMPS) and meso-2,3-dimercaptosuccinic acid (DMSA) are the treatments of choice for mercury toxicity. Alpha-lipoic acid (ALA), a disulfide, and its metabolite dihydrolipoic acid (DHLA), a dithiol, have also been shown to have chelation properties when used in an appropriate manner. Whilst N-acetyl-cysteine (NAC) and glutathione (GSH) have been recommended in the treatment of mercury toxicity in the past, an examination of available evidence suggests these agents may in fact be counterproductive. Zinc and selenium have also been shown to exert protective effects against mercury toxicity, most likely mediated by induction of the metal binding proteins metallothionein and selenoprotein-P. Evidence suggests however that the co-administration of selenium and dithiol chelation agents during treatment may also be counter-productive. Finally, the issue of diagnostic testing for chronic, historical or low dose mercury poisoning is considered including an analysis of the influence of ligand interactions and nutritional factors upon the accuracy of "chelation challenge" tests. PMID- 17408841 TI - Type I interferon response in the central nervous system. AB - This review is dedicated to the influence of type I IFNs (also called IFN alpha/beta) in the central nervous system (CNS). Studies in mice with type I IFN receptor or IFN-beta gene deficiency have highlighted the importance of the type I IFN system against CNS viral infections and non-viral autoimmune disorders. Direct antiviral effects of type I IFNs appear to be crucial in limiting early spread of a number of viruses in CNS tissues. Type I IFNs have also proved to be beneficial in autoimmune disorders like multiple sclerosis or experimental autoimmune encephalitis, probably through immunomodulatory effects. Increasing efforts are done to characterize IFN expression and response in the CNS: to identify type I IFN producing cells, to decipher pathways leading to type I IFN expression in those cells, and to identify responding cells. However, reversible and irreversible damages consecutive to chronic exposure of the CNS to type I IFNs underline the importance of a tightly regulated type I IFN homeostasis in this organ. PMID- 17408842 TI - Post-transcriptional control of the interferon system. AB - The interferon (IFN) system is a well-controlled network of signaling, transcriptional, and post-transcriptional processes that orchestrate host defense against microbes. The IFN response comprises a multi-array of IFN-stimulated gene products that mediate a variety of biological processes designed to control infection and regulate specific immune responses. In this review, we focus on post-transcriptional mechanisms of gene regulation that occur during the course of IFN induction and during the response of cells to IFN. Post-transcriptional mechanisms involve different levels of regulation such as mRNA stability, alternative splicing, and translation. Such controls offer a fine tuning mechanism for efficient and rapid response and as a negative feedback control in IFN biosynthesis and response. PMID- 17408843 TI - Ordering events of biochemical evolution. AB - Metabolic pathways exhibit structures resulting from an evolutionary process. Pathways have been inherited through time with modification, from the earliest periods of life. It is possible to compare the structure of pathways as done in comparative anatomy, i.e. for inferring ancestral pathways or parts of it (ancestral enzymatic functions), using standard phylogenetic reconstruction. Thus a phylogenetic tree of pathways provides a relative ordering of the rise of enzymatic functions. It even becomes possible to order the birth of each complete pathway in time. This particular "DNA-free" conceptual approach to evolutionary biochemistry is reviewed, gathering all the justifications given for it. Then, the method of assigning a given pathway to a time span of biochemical development is revisited. The previous method used an implicit "clock" of metabolic development that is difficult to justify. We develop a new clock-free approach, using functional biochemical arguments. Results of the two methods are not significantly different; our method is just more precise. This suggests that the clock assumed in the first method does not provoke any important artefact in describing the development of biochemical evolution. It is just unnecessary to postulate it. As a result, most of the amino acid metabolic pathways develop forwards, confirming former models of amino acid catabolism evolution, but not those for amino acid anabolism. The order of appearance of sectors of universal cellular metabolism is: (1) amino acid catabolism, (2) amino acid anabolism and closure of the urea cycle, (3) glycolysis and glycogenesis, (4) closure of the pentose-phosphate cycle, (5) closure of the Krebs cycle and fatty acids metabolism, (6) closure of the Calvin cycle. PMID- 17408844 TI - The human 2'-5'oligoadenylate synthetase family: unique interferon-inducible enzymes catalyzing 2'-5' instead of 3'-5' phosphodiester bond formation. AB - The demonstration by Kerr and colleagues that double-stranded (ds) RNA inhibits drastically protein synthesis in cell-free systems prepared from interferon treated cells, suggested the existence of an interferon-induced enzyme, which is dependent on dsRNA. Consequently, two distinct dsRNA-dependent enzymes were discovered: a serine/threonine protein kinase that nowadays is referred to as PKR and a 2'-5'oligoadenylate synthetase (2'-5'OAS) that polymerizes ATP to 2'-5' linked oligomers of adenosine with the general formula pppA(2'p5'A)(n), n>or=1. The product is pppG2'p5'G when GTP is used as a substrate. Three distinct forms of 2'-5'OAS exist in human cells, small, medium, and large, which contain one, two, and three OAS units, respectively, and are encoded by distinct genes clustered on the 2'-5'OAS locus on human chromosome 12. OASL is an OAS like IFN induced protein encoded by a gene located about 8 Mb telomeric from the 2'-5'OAS locus. OASL is composed of one OAS unit fused at its C-terminus with two ubiquitin-like repeats. The human OASL is devoid of the typical 2'-5'OAS catalytic activity. In addition to these structural differences between the various OAS proteins, the three forms of 2'-5'OAS are characterized by different subcellular locations and enzymatic parameters. These findings illustrate the apparent structural and functional complexity of the human 2'-5'OAS family, and suggest that these proteins may have distinct roles in the cell. PMID- 17408845 TI - Down syndrome gene dosage imbalance on cerebellum development. AB - Down syndrome (DS) is a chromosomal disorder whereby genes on chromosome 21 are present in three copies. This gene copy imbalance is thought to be responsible for a number of debilitating conditions experienced by individuals with DS. Amongst these is a reduced cerebellar volume, or cerebellar hypoplasia, which is believed to contribute to the perturbation of fine motor control. Mouse models of DS (such as Ts65Dn, Ts1Cje, Tc1) exhibit a cerebellar phenotype similar to that of individuals with DS and which primarily manifests as a disruption of the density of the granule cell layer. Dissecting which of the three-copy genes are responsible for this phenotype (the primary gene dosage effect) has been a task undertaken by researchers working with various segmental trisomies and transgenic mice. It is generally agreed that, when expressed, three-copy genes of trisomic mice are expressed at around 1.5 times that of the same genes in euploid (wild type) mice. However, amongst these studies there does not appear to be a consensus on the nature and extent of differential expression of two-copy genes in trisomic mice-the secondary dosage effect. Much of this variation may have to do with the stage of development investigated and the nature and complexity of the tissue (i.e. whole brain versus the cerebellum). The recent discovery that trisomic granule cell precursors are less sensitive to sonic hedgehog-induced proliferation has opened up another avenue for the identification of three-copy genes responsible for the cerebellar phenotype. It is hoped that further investigation of this phenomenon, together with new mouse segmental trisomies and transgenics, will reveal the cause of the proliferation deficit and allow for potential treatment. PMID- 17408846 TI - The risk of shoulder dystocia related permanent fetal injury in relation to birth weight. AB - OBJECTIVE: To examine birth weight related risks of fetal injury in connection with shoulder dystocia. STUDY DESIGN: The investigation was based on a retrospective analysis of 316 fetal neurological injuries associated with deliveries complicated by arrest of the shoulders that occurred across the United States. RESULTS: The study revealed that the distribution of birthweights for the high risk shoulder dystocia population differs from the standard birthweight distribution. The relative difference per birthweight interval is used to adjust an assumed 1:1000 baseline risk of injury due to shoulder dystocia following vaginal deliveries. These adjusted risks show a need to consider new thresholds for elective cesarean delivery. CONCLUSIONS: Current North American and British guidelines, that set 5000 g as minimum estimated fetal weight limit for elective cesarean section in non-diabetic and 4500 g for diabetic gravidas, may expose some macrosomic fetuses to a high risk of permanent neurological damage. The authors present the opinion that the mother, having been informed of the risks of vaginal versus abdominal delivery, should be allowed to play an active role in the critical management decisions. PMID- 17408847 TI - Coliforms removal in full-scale activated sludge plants in India. AB - This paper investigates the removal of coliforms in full-scale activated sludge plants (ASP) operating in northern regions of India. Log2.2 and log2.4 removal were observed for total coliforms (TC) and fecal coliforms (FC), respectively. However, the effluent still contained a significant number of TC and FC which was greater than the permissible limit for unrestricted irrigation as prescribed by WHO. The observations also suggest that extended aeration (EA) process operating under high mixed liquor suspended solids (MLSS) and sludge retention time (SRT) is more efficient in the removal of coliforms. Further attempts have been made to establish the relationship between two key wastewater parameters, i.e. biochemical oxygen demand (BOD) and suspended solids (SS) with respect to fecal and TC. The relationships were observed to be linear with a good coefficient of correlation. The interrelationship of BOD and SS with coliforms manifest that improvement of the microbiological quality of wastewater could be linked with the removal of SS. Therefore, SS can serve as a regulatory tool in lieu of an explicit coliforms standard. PMID- 17408848 TI - Passerini-glazel feminizing genitoplasty: modifications in 17 years of experience with 82 cases. AB - OBJECTIVES: To describe modifications of Passerini-Glazel feminizing genitoplasty and report on long-term functional outcome. METHODS: Modifications include vaginal dissection and disconnection from the urethrovaginal sinus as the initial stage of the procedure; large dissection of the neurovascular bundle on both dorsal and lateral faces of the clitoris; plication of the skin around the reduced clitoris; and suturing the lateral edge of the proximal portion of the mucocutaneous plate with the labia majora's medial edge to a plane deeper than the subcutaneous tissue. These modifications reduce bleeding and operating time, better preserve clitoral sensitivity, form the clitoral prepuce, and create labia minora. RESULTS: Eighty-two patients underwent modified Passerini-Glazel feminizing genitoplasty. Mean operating time was 120min (range: 100-180). Forty six patients (46 of 82, 56%) were assessed at a mean follow-up of 5 yr (range: 2 9). There were no cases of clitoral vascularization defect or urethrovaginal fistula. The urethral meatus was never hypospadic. The vaginal introitus was large and elastic in all cases. Vaginal caliber at the internal suture line was as large as the vaginal introitus and the distal native vagina in 20 (43.5%) of the 46 girls. All mothers and patients reported satisfaction with external genital appearance. CONCLUSIONS: These long-term results suggest that our modifications of one-stage Passerini-Glazel feminizing genitoplasty facilitate the procedure and produce good cosmetic results. PMID- 17408849 TI - Segmental testicular infarction: conservative management is feasible and safe. AB - Segmental testicular infarction is a rare cause of acute scrotum. Its aetiology is not well defined and it can be clinically confused with a testicular tumour. Because the differential diagnosis between segmental testicular infarction and testicular tumour can be difficult, most authors in the past recommended surgery. Imaging plays an important role in the preoperative diagnosis, with a colour Doppler ultrasonography as the investigation of choice although magnetic resonance imaging (MRI) can be useful in doubtful cases. The objective of this retrospective study was to describe the radiologic findings and the outcome of conservative management in a single-centre experience of 19 cases of segmental testicular infarction. PMID- 17408850 TI - Renal cell carcinoma guideline. AB - OBJECTIVES: The European Association of Urology (EAU) Guideline Group for renal cell carcinoma (RCC) prepared this guideline to help urologists assess the evidence-based management of RCC and to incorporate the guideline recommendations into their clinical practice. METHODS: The recommendations provided in the current guideline are based on a systematic literature search using MedLine, the Cochrane Central Register of Controlled Trials, and publications and review articles. RESULTS: A limited number of prospective randomised studies are available with high-level evidence. Most publications concerning RCC are based on retrospective analyses, including some larger multicentre validation studies and well-designed controlled studies. CONCLUSIONS: It must be stressed that the current guideline contains information for the treatment of an individual patient according to a standardised general approach. Updated recommendations concerning diagnosis, treatment, and follow-up can improve the clinical handling of patients with RCC. PMID- 17408851 TI - Paradoxical fixed dilatation of the contralateral pupil as a false-localizing sign in intraparenchymal frontal hemorrhage. AB - Unilateral fixed pupillary dilatation represents an important clinical sign of transtentorial herniation of an ipsilateral mass lesion. Rarely the contralateral pupil is affected to produce a false-localizing sign. Two cases of this paradoxical contralateral fixed pupillary dilatation involving intra-axial lesions have been reported. We report a case of a 33-year-old man with a left frontal intraparenchymal hemorrhage who developed a false localizing fixed dilatation of his right pupil, which resolved after craniotomy and evacuation of the clot. The possible mechanisms of contralateral third nerve palsy are discussed. PMID- 17408852 TI - Meningitis caused by Capnocytophaga canimorsus: when to expect the unexpected. AB - In this article we review the available data concerning meningitis caused by Capnocytophaga canimorsus. The clinical presentation of this rare condition is described with the emphasis on associated conditions and management issues. Two additional cases, illustrating the difficulties in recognizing this rare disease, are presented. Reviewing a total of 28 reported cases, a preceding bite-incident by a cat or dog, or close contact with these animals, was described in the majority of cases (89%). Patients had a median age of 58 years; splenectomy and alcohol abuse were noted in, respectively, 18% and 25% of patients. Only in one case immune suppressive drug use was reported. The diagnosis C. canimorsus meningitis should be considered in healthy and immunocompromised adults, especially after splenectomy, who present with symptoms attributable to meningitis and a history of recent exposure to dogs or cats. The possibility of this condition has implications for both the diagnostic work-up and the treatment of the patient. PMID- 17408853 TI - Assessment of a new carbon tetrachloride destruction system based on a microwave plasma torch operating at atmospheric pressure. AB - A new system for destroying volatile organic waste based on a microwave plasma torch that operates at atmospheric pressure and is coupled to a reactor affording isolation of output gases and adjustment of the plasma discharge atmosphere is proposed. The system was assessed by using carbon tetrachloride as the target volatile organic compound (VOC) and argon as the main gas in a helium atmosphere. Under optimal conditions, a microwave power of less than 1000 W was found to reduce the CCl(4) concentration at the reactor outlet to the parts-per-billion level and hence to virtually completely destroy the VOC. With high argon flow rates and CCl(4) concentrations, the energy efficiency can reach levels in excess of 3000 g/kWh. Output gases and species in the plasma, which were identified by gas chromatography and light emission spectroscopy, respectively, were found to include no halogen-containing derivatives resulting from the potential cleavage of CCl(4). In fact, the main gaseous byproducts obtained were CO(2), NO and N(2)O, in addition to small traces of Cl(2), and the solid byproducts Cl(2)Cu and various derivatives depending on the particular reactor zone. PMID- 17408854 TI - Influence of delayed CSF storage on concentrations of phospho-tau protein (181), total tau protein and beta-amyloid (1-42). AB - It is generally accepted that cerebrospinal fluid (CSF) biomarkers such as tau protein, phosphorylated tau protein (threonine 181) and beta-amyloid (1-42) can facilitate early and differential diagnosis of Alzheimer's disease (AD). Since the respective concentrations can only be measured in a number of specialized centers, time to CSF specimen work-up has been considered as crucial for the stability of the respective biomarkers. When shipping of CSF samples is needed for biomarker measurement and immediate freezing of samples is not available, an overnight delay of up to 24h frequently occurs. Therefore, we investigated the potential impact of a 24h delayed freezing on CSF biomarker concentrations and compared it to 2h storage (room temperature, 20 degrees C) and an immediate freezing. First, storage at room temperature for 2h had only marginal, non significant effects on the concentrations of CSF total tau protein and phospho tau protein (181) compared to immediate freezing. Second, storage at room temperature for 24h did not significantly affect total tau protein or phospho-tau protein but beta-amyloid (1-42) concentrations which increased significantly compared to the samples frozen immediately. These results indicate that CSF samples for the evaluation of total tau and phospho-tau protein may be kept at room temperature for up to 24h whereas CSF samples for beta-amyloid (1-42) need to be frozen immediately. PMID- 17408855 TI - Spatial coincidence modulates the Colavita visual dominance effect. AB - The Colavita visual dominance effect refers to the phenomenon whereby participants presented with auditory, visual, or audiovisual stimuli in a speeded response task sometimes fail to respond to the auditory component of the bimodal targets. We conducted an experiment on the Colavita effect in which the auditory and visual components of the bimodal targets were presented from either the same or different positions (sides) at one of two eccentricities (13 degrees or 26 degrees ). Participants were presented with auditory, visual, and bimodal stimuli to which they had to respond by pressing an auditory response key, a visual response key, or both response keys, respectively. On bimodal trials, participants failed to respond to the auditory stimulus significantly more often than they failed to respond to the visual stimulus, resulting in a significant Colavita visual dominance effect. The Colavita effect was significantly larger when the stimuli were presented from the same position than when they were presented from different positions. These results provide the first empirical evidence that the Colavita effect is modulated by the spatial coincidence of the auditory and visual stimuli. PMID- 17408856 TI - D-serine relieves chronic lead exposure-impaired long-term potentiation in the CA1 region of the rat hippocampus in vitro. AB - Chronic lead-exposure produces long-lasting astroglial morphological and functional changes, which disturb the neuronal functions in the hippocampus. It has been shown that glia-derived D-serine is an essential signal for N-methyl-D aspartate receptor (NMDAR)-dependent synaptic plasticity in the hippocampal CA1 region. However, the relationship between d-serine and the chronic lead exposure induced deficit of synaptic plasticity is not clear. In the present study, the properties of D-serine on the chronic lead exposure-impaired synaptic plasticity in the rat hippocampal CA1 region were investigated with electrophysiological recording techniques in vitro. We found that 50 microM D-serine rescued the chronic lead exposure-induced deficit of long-term potentiation (LTP). However, this effect could be abolished by 7-chlorokynurenic acid (7-ClKY), which is a specific antagonist of the glycine-binding site of NMDARs. In contrast, D-serine had no effect on the NMDAR-independent LTP, which was induced in the mossy-CA3 synapses. In addition, we found that d-serine rescued the acute Pb(2+)-impaired NMDAR-mediated excitatory postsynaptic currents (EPSCs) partially. These findings demonstrate that d-serine relieves the chronic lead exposure-induced deficit of synaptic plasticity via NMDAR activation suggesting that administration of d serine may be a potential therapeutic intervention to treat chronic lead exposure impaired cognitive functions or affective disorders. PMID- 17408857 TI - Iron, copper, and iron regulatory protein 2 in Alzheimer's disease and related dementias. AB - Accumulating evidence implicates a role for altered iron and copper metabolism in the pathogenesis of neurodegenerative disorders such as Alzheimer's disease (AD). However, imbalances in the levels of the various forms of iron at different stages of AD have not been examined. In this pilot study we extracted and measured the levels of loosely bound, non-heme and total iron and copper in the frontal cortex and hippocampus of patients with mild-moderate AD (n=3), severe AD (n=8) and dementia with Lewy bodies (DLB, n=6), using graphite furnace atomic absorption spectrometry (GFAAS). Additionally, the expression of iron regulatory protein 2 (IRP2) was examined in relation to the pathological hallmarks of AD and DLB, amyloid plaques, neurofibrillary tangles (NFT), and Lewy bodies, by immunohistochemistry. We found significantly decreased loosely bound iron in the hippocampal white matter of mild-moderate and severe AD patients and a trend towards increased non-heme iron in the hippocampal gray matter of severe AD patients. Furthermore, decreased levels of total copper were seen in severe AD and DLB frontal cortex compared to controls, suggesting an imbalance in brain metal levels in both AD and DLB. The decrease in loosely bound iron in mild moderate AD patients may be associated with myelin breakdown seen in the beginning stages of AD and implicates that iron dysregulation is an early event in AD pathogenesis. PMID- 17408858 TI - The role of catastrophizing in sickle cell disease--the PiSCES project. AB - In several types of chronic pain, catastrophizing has been related to higher pain intensity, and health care utilization but it has not been explored extensively in sickle cell disease (SCD). The objective of the study was to identify the role of catastrophizing in SCD, specifically in relation to painful crises, non-crisis pain, and responses to pain. Two hundred and twenty SCD adults were enrolled in a prospective cohort study of pain and completed between 30 and 188 daily diaries in 6 months. The Catastrophizing subscale (CAT) of the Coping Strategy Questionnaire (CSQ) was administered at baseline and at study exit. Depression and quality of life were measured by the Patient Health Questionnaire and SF-36, respectively, at baseline. The CAT mean was 13.6 (SD=8.4) and higher CAT was correlated with greater depression severity (r=0.48; p<0.001) and poorer quality of life in all domains (r=-0.24 to -0.47; p<0.001). There was no significance difference between CAT mean baseline and exit scores, and the measures were strongly correlated within patients (r=0.69; p<0.001). No difference was found between higher and lower catastrophizers in intensity of pain, distress, interference, and health service utilization, both on crisis or non-crisis SCD related pain days, after controlling for depression. Adults with SCD had a higher mean catastrophizing score than found in studies of other chronic pain conditions that are not lifelong and life-threatening. CAT scores were not correlated with pain parameters or utilization. The role of catastrophizing in other conditions cannot be generalized to SCD. PMID- 17408860 TI - Improving the emission characteristics of a carbon nanotube cathode in an aging process. AB - Aging of the field emission performance of the printed carbon nanotubes (CNTs) cathode is studied. A continuous increase of the field emission current as well as the density of field emission sites under a constant voltage is observed. It is revealed that the resistant heating may play an important role in the activation of the potential emitters during the aging process. A technique of activating the printed CNT cathode with an aging process is suggested. F-N curves before and after the aging processes are analyzed and it is revealed that the effective emission area increases during the aging process. PMID- 17408861 TI - The effect of anticoagulants on cancer risk and survival: systematic review. AB - OBJECTIVES: Several in vitro and in vivo studies have shown that low molecular weight heparin and warfarin may directly inhibit tumour cell growth and prevent metastatic spread. However, the clinical evidence in support of an anti-cancer effect is less conclusive. We summarize the evidence from clinical studies that examine the effect of these anticoagulants on cancer development and briefly describe the current understanding of the potential mechanisms by which anticoagulants may exert an anti-cancer effect. METHODS: English-language articles reporting on warfarin, coumarin or low molecular weight heparin for the treatment or prevention of cancer were selected from PUBMED. All randomized clinical trials, case-control studies, cohort studies, and meta-analyses were retrieved. Detailed data review and abstraction was performed according to pre specified criteria. RESULTS: Of ninety-nine articles retrieved, 12 warfarin and 17 low molecular weight heparin articles were included in the review. We found no consistent evidence that warfarin may improve cancer survival, though there is indirect evidence that prolonged warfarin use may decrease the risk of urogenital cancer. Low molecular weight heparin may improve survival of patients with small cell lung cancer and those with advanced malignancy who have more favorable prognoses. CONCLUSION: Clinical evidence exists in support of an anti-neoplastic effect of anticoagulants. However, more research is needed to further define which cancer type and stage would most benefit from low molecular weight heparin, as well as to explore the role of warfarin in urogenital tumour development. PMID- 17408862 TI - Strain-specific differences in the effects of cyclosporin A and FK506 on the survival and regeneration of axotomized retinal ganglion cells in adult rats. AB - The immune response can influence neuronal viability and plasticity after injury, effects differing in strains of rats with different susceptibility to autoimmune disease. We assessed the effects of i.p. injections of cyclosporin A (CsA) or FK506 on adult retinal ganglion cell (RGC) survival and axonal regeneration into peripheral nerve (PN) autografted onto the cut optic nerve of rats resistant (Fischer F344) or vulnerable (Lewis) to autoimmune disease. Circulating and tissue CsA and FK506 levels were similar in both strains. Three weeks after autologous PN transplantation the number of viable beta-III tubulin-positive RGCs was significantly greater in CsA- and FK506-treated F344 rats compared with saline-injected controls. RGC survival in Lewis rats was not significantly altered. In F344 rats, retrograde labeling of RGCs revealed that CsA or FK506 treatment significantly increased the number of RGCs that regenerated an axon into a PN autograft; however these agents had no beneficial effect on axonal regeneration in Lewis rats. PN grafts in F344 rats also contained comparatively more pan-neurofilament immunoreactive axons. In both strains, 3 weeks after transplantation CsA or FK506 treatment resulted in increased retinal macrophage numbers, but only in F344 rats was this increase significant. At this time-point PN grafts in both strains contained many macrophages and some T cells. T cell numbers in Lewis rats were significantly greater than in F344 animals. The increased RGC axonal regeneration seen in CsA- or FK506-treated F344 but not Lewis rats shows that modulation of immune responses after neurotrauma has complex and not always predictable outcomes. PMID- 17408863 TI - Exploration of prostanoid receptor subtype regulating estradiol and prostaglandin E2 induction of spinophilin in developing preoptic area neurons. AB - The prostaglandin E2 (PGE2) mediates estradiol-induced masculinization of sexual behavior in the rat during a perinatal sensitive period. PGE2 induces formation of dendritic spines on preoptic area (POA) neurons and this synaptic pattern change is associated with the ability to express male sexual behavior as an adult. Whether PGE2 is released from astrocytes or neurons in the developing POA is unknown. To further understanding of how PGE2 induces dendritic spine formation at the cellular level, we have explored the PGE2 receptor subtype mediating this response. There are four receptors for PGE2, EP1, EP2, EP3 and EP4, each having unique but interacting signal transduction profiles. Treatment of newborn female rats with the EP receptor agonists iloprost, butaprost and sulprostone indicated that stimulation of both the EP2 and EP3 receptors significantly increased spinophilin, a protein whose levels positively correlate to the presence of dendritic spines and masculinization of the POA. Use of antisense oligonucleotides against the mRNA for each receptor reveals that either EP2 or EP3 receptor knockdown reduces spinophilin in PGE2- or estradiol-treated females, whereas reducing EP1 or EP4 receptor levels by the same means has a smaller but also significant effect. A developmental profile of EP receptor expression indicates EP1 in particular is elevated for the first few days of life, corresponding to the critical period for masculinization, whereas mRNA levels for the other three receptors remain relatively constant. PMID- 17408864 TI - Expression and immunolocalization of the plasma membrane monoamine transporter in the brain. AB - High affinity monoamine transporters efficiently terminate neurotransmission through synaptic reuptake of released neurotransmitter. We recently cloned and characterized a novel low-affinity, high capacity plasma membrane monoamine transporter (PMAT) that is strongly expressed in the human brain and efficiently transports 5-HT and dopamine (DA). In efforts to understand the physiological function of PMAT and its relevance in monoaminergic pathways, we cloned the PMAT homolog from the mouse brain, demonstrated its capability for transporting 5-HT and DA, and determined the regional and cellular localization of mouse plasma membrane monoamine transporter (mPMAT) in adult mouse brain by reverse transcription polymerase chain reaction, non-radioactive in situ hybridization, and immunohistochemical methods. Our results showed that mPMAT mRNA and protein are broadly expressed in the mouse brain and are particularly abundant in forebrain cortex, olfactory tubercle, hippocampus, cerebellum and epithelial cells of the choroid plexus. Dual-immunofluorescence histochemistry with established phenotypic markers microtubule-associated protein (MAP2) and glial fibrillary acidic protein (GFAP) revealed that mPMAT is expressed in neuronal cells but not in astrocytes. mPMAT is co-expressed in many brain regions with the high affinity 5-HT transporter (SERT) and the dopamine transporter (DAT), but is also found in certain sites that receive monoamine innervation but lack significant expression of SERT or DAT. These findings suggest that mPMAT is a widely distributed, neuronally-expressed transporter, which may support the role of 5-HT and DA uptake under certain conditions. PMID- 17408865 TI - Is the cortisol awakening rise a response to awakening? AB - A distinct rise in cortisol levels that occurs after morning awakening is increasingly used as an indicator of adrenocortical activity which is associated with different pathologies. Although it was previously assumed that the transition from sleep to wake is essential for the occurrence of the cortisol morning rise, this has never been tested. Here, we examined 16 healthy young men (20-33 yrs) between 2300 and 0800 h under sleep laboratory conditions. Serum cortisol and plasma adrenocorticotropin (ACTH) as well as salivary cortisol levels (after subjects were woken up at 0700 h) were repeatedly assessed. In a supplementary study condition, salivary cortisol levels in the first hour after awakening were measured at the subjects' home on two consecutive days. Comparison of pre- and post awakening measurements revealed significantly steeper increases in cortisol and ACTH after awakening. The rise in cortisol upon awakening under laboratory conditions did not significantly differ from that observed at home. We conclude that the cortisol increase after awakening is a response to morning awakening that is distinct from the circadian rise in hypothalamo-pituitary adrenal (HPA) activity in the morning hours. Although the cortisol awakening response is modulated by circadian influences, it primarily reflects phasic psychophysiological processes specific to the sleep-wake transition. PMID- 17408866 TI - Behavioral Activation System (BAS) sensitivity and reactivity to alcohol cues among hazardous drinkers. AB - Previous research has suggested that Behavioral Activation System (BAS) sensitivity may be associated with stronger appetitive responses to alcohol cues. This study was conducted to explore whether those with higher BAS sensitivity showed greater urge and affective responses to alcohol cues and whether different types of appetitive cues moderated the magnitude of these associations. One hundred eighty-eight hazardous drinkers (90 women) were exposed to the sight and smell of their favorite alcoholic beverage during a cue exposure procedure. Participants were asked to either lean towards the beverage (Cue Only) when signaled by tones, or lift the beverage towards them (Cue+Action). BAS sensitivity was significantly associated with baseline ratings of urge and affect, and was found to be a significant predictor of urge and affect reactivity; however significant interaction effects with cue type were not observed. Results provide further evidence for the influence of individual differences in reward responsiveness on alcohol use and abuse. PMID- 17408867 TI - Impact of positive and anxious mood on implicit alcohol-related cognitions in internally motivated undergraduate drinkers. AB - We explored the impact of musically induced positive and anxious mood on the implicit alcohol-related cognitions of 48 undergraduate students who drink either to enhance positive mood states (EM) or to cope with anxiety (CM-anxiety). Participants completed a post-mood induction computerized alcohol Stroop task that taps implicit alcohol-related cognitions. As hypothesized, CM-anxiety participants in the anxious (but not those in the positive) mood condition showed longer colour-naming latencies for alcohol (vs. non-alcohol) targets (i.e., an attentional bias toward alcohol-related stimuli). Also conforming to expectation was the finding that EM participants in the positive (but not those in the anxious) mood condition displayed longer colour-naming latencies for (i.e., an attentional bias toward) alcohol (vs. non-alcohol) target words. PMID- 17408868 TI - History of smoking and postcessation weight gain among weight loss surgery candidates. AB - Smoking cessation often results in weight gain. Although smoking cessation frequently is recommended to patients presenting for weight loss surgery (WLS), the relationship between smoking cessation and weight gain among WLS candidates is poorly understood. Thus, we sought to document the history and prevalence of smoking and smoking-related weight gain among WLS candidates. Subjects (N=67) presenting for bariatric surgery provided demographic information, were interviewed about smoking, and were weighed and measured prior to operation. Sixty-seven percent of patients reported a lifetime history of smoking, and 26.9% were current smokers. Among lifetime smokers who had attempted to quit, the average maximum amount of weight gained following smoking cessation was 28.1 lb, but there was wide variability in postcessation weight gain. These data suggest that smoking among candidates for bariatric surgery is prevalent and that previous cessation attempts were associated with considerable weight gain. Because patients often receive recommendations to quit smoking and lose weight prior to surgery, additional information on the impact of presurgical smoking cessation on long-tem weight control in this population is needed. PMID- 17408869 TI - The motivation to stay abstinent in ex-smokers: comparing the present with the past. AB - Little is known about the motivation of ex-smokers to stay abstinent. In the present study we argue that ex-smokers compare their present to their past when they still smoked to conclude whether they make good progress towards a satisfactory state of continued abstinence. These temporal comparisons are thought to be central in the motivation to stay abstinent in ex-smokers. The power of temporal comparisons to predict relapse was tested in two related samples of ex-smokers (N=152 and N=197), together with two other relevant psychological factors; positive outcome expectations of smoking and self-efficacy expectations. In the first sample of ex-smokers, only temporal comparisons predicted relapse after 2 months. In the second sample of ex-smokers, temporal comparisons mediated the relation between perceived positive outcomes of smoking and relapse after 6 months. In addition, in predicting relapse after 6 months, temporal comparisons interacted with self-efficacy. The present study suggests that temporal comparisons comprise the cognitive aspect of the motivation of ex smokers to stay abstinent. This conceptualization of the motivation in ex-smokers can be used in practice to prevent relapse. PMID- 17408870 TI - Personality risk profile for conduct disorder and substance use disorders in youth. AB - The five factor model of personality is a useful metric to describe personality profiles associated with maladaptive functioning. Using the NEO-Five Factor Inventory (NEO-FFI), we examined a conceptually based profile of high neuroticism, low agreeableness and low conscientiousness among 243 youth (aged 13 18 years) with varying degrees of conduct disorder (CD) and substance use disorders (SUD). Comparisons of the NEO-FFI personality dimensions between CD/SUD youth and adolescent siblings (N=173), and relations between the personality dimensions and behavioral indicators of conduct disorder and substance involvement were examined. Youth with CD and SUD had greater neuroticism, lower agreeableness, and lower conscientiousness than siblings of a similar age. The NEO-FFI scales predicted aggression and substance involvement for both probands and siblings in this cross-sectional investigation. These findings support the role for personality in models of the etiology and persistence of conduct disorder and substance use disorders. PMID- 17408871 TI - Acculturation, gender, and alcohol use among Mexican American college students. AB - Prior research with non-college samples of Mexican Americans has demonstrated that gender moderates the association between acculturation and alcohol use. We replicated this finding in a college student sample and attempted to account for the differential impact of acculturation on Mexican American men and women by examining the mediating effects of social context, family conflict and psychological functioning. Participants were 148 Mexican Americans (67% female; M age 23 years) from three state universities in California and Texas who completed self-report surveys. In multivariate analyses controlling for age, maternal education, living situation, and site, linguistic acculturation was associated with increased alcohol use and misuse among women but not men. Two social context variables (social facilitation and family drinking) mediated the association between acculturation and alcohol use (heavy drinking, past year alcohol use, and a composite drinking variable) among women. The findings highlight the importance of social context for understanding alcohol use by Latina college students and indicate directions for future research and intervention development. PMID- 17408872 TI - A potential therapeutic strategy for capillary hemangioma with new anti-vascular endothelial growth factor (anti-VEGF) agents. PMID- 17408873 TI - Thiazolidinediones: from antioxidant to neurotherapeutic? AB - A prevailing paradigm regarding the pathophysiology of mood disorders posits that these syndromes are possibly neurodegenerative. Alterations in interacting biological networks which subserve metabolism, inflammation, immune function, and stress response are hypothesized to mediate the neurotoxicity and allostasis associated with mood disorders. Preclinical evidence indicates that thiazolidinediones (TZDs) exert neurotherapeutic (e.g., neurotrophic) effects. Preliminary clinical evidence also suggests that TZDs may be salutary for mental disorders in which neurocognitive deficits are a central feature. We propose that TZDs constitute a potentially novel disease-modifying treatment avenue for mood disorders. PMID- 17408874 TI - Anemia of immobility: caused by adipocyte accumulation in bone marrow. AB - Anemia of chronic disease has long been used to classify a non-regenerative, low grade, chronic, normocytic, normochromic anemia that presents with no obvious etiology. Within this group, some patients have a chronic inflammatory condition that limits erythrocyte generation or access to iron stores. This specific type of anemia has been termed anemia of chronic inflammation. However, a substantial remainder of patients diagnosed with anemia of chronic disease present with no active inflammation. These include many clinical populations with reduced limb loading, such as spinal cord injured patients, astronauts, elderly people with limited mobility and experimental bed-rest subjects. In some populations with decreased mobility, accumulation of fat in the bone marrow has been demonstrated. We hypothesize that adipocyte accumulation in bone marrow both passively and actively impairs erythropoiesis and thus defines a new type of anemia called anemia of immobility. The non-specific umbrella term anemia of chronic disease thus becomes obsolete in favour of either the diagnosis of anemia of immobility or anemia of chronic inflammation according to the distinct mechanism involved. PMID- 17408875 TI - [Extra-uterine pelvic leiomyoma: diagnosis and practical management]. AB - OBJECTIVES: To analyze the possibilities of setting up a therapy for extra uterine pelvic leiomyomas. METHODS: Three cases of leiomyomas of the broad ligament, of the round ligament and of the ovary, and literature review. RESULTS: Little is known about physiopathology of extra-uterine leiomyoma. The diagnosis of extra-uterine leiomyoma is based on histopathological analysis, using standard histology, and immunohistochemistry with anti-desmin and anti smooth muscle actin antibodies. The main differential diagnoses are fibroma, fibrothecoma, ovarian fibrosarcoma, and gastrointestinal stromal tumors. To define criteria of malignancy, we use Bell's classification without being sure that the uterine and extra-uterine models are comparable. So there is a risk of ignoring a low grade leiomyosarcoma. Providing therapy depends on the clinicopathologic features: the so called "parasitic leiomyoma", a tumor developed at the expense of local smooth muscle cells, metastasis of a benign metastasizing leiomyoma or leiomyomatosis peritonealis disseminata. CONCLUSION: The extra-uterine leiomyoma has no precise nosologic status and no specific criteria of benignity; thus no precise evolution can be predicted. We must be extremely careful, and the issue of the monitoring and long-term therapy of patients must come up. PMID- 17408876 TI - [Carcinosarcoma of the ovary]. AB - Ovarian carcinosarcoma, also called malignant mixed mesodermal tumour, is a rare ovarian tumour representing less than two per cent of ovarian cancers. Carcinosarcoma is an aggressive tumour, which associates some epithelial elements (carcinoma) with a stromal component (sarcoma). This tumour can be found in the female genital tractus, mostly in the uterus. It can be found even more rarely on the ovaries. The initial stage of the disease at the diagnostic is considered as the only prognostic factor. There is no existing consensus concerning treatment. Nevertheless, surgical treatment is paramount for the survival of patients. Response rates to chemotherapy are about 20%. PMID- 17408877 TI - The impact of visual perception on equine learning. PMID- 17408878 TI - Equine learning behaviour: accounting for ecological constraints and relationships with humans in experimental design. PMID- 17408879 TI - Equine learning in a wider context--opportunities for integrative pluralism. PMID- 17408880 TI - Equine learning behaviour: limits of ability and ability limits of trainers. PMID- 17408881 TI - Learning about horses: what is equine learning all about? PMID- 17408882 TI - Genome-wide analysis of the auxin response factors (ARF) gene family in rice (Oryza sativa). AB - Auxin response factors (ARFs) are transcription factors that bind with specificity to TGTCTC-containing auxin response elements (AuxREs) found in promoters of primary/early auxin response genes and mediate responses to the plant hormone auxin. The ARF genes are represented by a large multigene family in plants. A comprehensive genome-wide analysis was carried out in this study to find all ARFs in Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa subsp. japonica), 23 and 25 ARF genes, named as AtARFs and OsARFs, were identified, respectively. Chromosomal locations of all OsARFs were presented and it was found that the duplication of OsARFs was associated with only the chromosomal block duplications but not local tandem duplications. A phylogenetic tree was generated from alignments of the full-length protein sequences of 25 OsARFs and 23 AtARFs to examine the phylogenetic relationships of rice and Arabidopsis ARF proteins. All 48 members of ARF gene families fell into three major classes, a total of 13 sister pairs, including 9 OsARF-OsARF, 2 AtARF-AtARF and 2 AtARF-OsARF sister pairs were formed, showing different orthologous relationships between AtARFs and OsARFs. EST analysis and RT-PCR assays demonstrated that 24 of all 25 OsARF genes were active and the transcript abundance of some OsARF genes was affected by auxin treatment or light- and dark-grown conditions. The outcome of the present study provides basic genomic information for the rice ARF gene family and will pave the way for elucidating the precise role of OsARFs in plant growth and development in the future. PMID- 17408883 TI - Functional characterization of the TMLH gene: promoter analysis, in situ hybridization, identification and mapping of alternative splicing variants. AB - Carnitine is a molecule with well-documented pleiotropic functions whose biosynthesis involves four catalytic steps. Here, we report a detailed analysis of the expression and transcriptional control of TMLH gene, which codifies for the first enzyme of carnitine biosynthesis. TMLH maps at the extreme end of Xq28, a chromosomal region of high genomic instability. By 5' and 3' RACE, we identified and mapped two alternative 5' TMLH first exons and seven alternative 3'-splice variants, which are spread over a genomic region of about 250 kb. While the two alternative 5' exons have different expression profiles, all the 3' alternative forms are ubiquitously expressed. Reporter assays revealed that the 3'-UTRs of each TMLH isoform might influence its own expression at post transcriptional level. In addition, we identified a highly conserved promoter region of TMLH. Functional analysis of this region showed the presence of a CpG island, whose methylation-status could control the level of TMLH transcription. Finally, by mRNA in situ hybridization, we found that TMLH expression is present at E12.5 dpc in the mouse liver, lung and brain, and is then maintained in the postnatal brain with a specific neuronal pattern. Collectively, our data highlight a tight transcriptional and post-transcriptional control of TMLH expression. PMID- 17408884 TI - Computational identification of microRNAs and their targets in Gossypium hirsutum expressed sequence tags. AB - MicroRNAs (miRNAs) are a class of non-coding RNAs that regulate gene post transcriptional expression in animals and plants. Comparatively genomic computational methods have been developed to predict new miRNAs in worms, humans, and Arabidopsis. Here we present an EST (Expressed Sequence Tags)--and GSS (Genomic Survey Sequences)-based combined approach for the detection of novel miRNAs in Gossypium hirsutum. This was initiated by using previously known miRNA sequences from Arabidopsis, rice and other plant species and an algorithm called miRNAassist to blast the databases of G. hirsutum EST and GSS. A total of 37 potential miRNAs were detected following a range of filtering criteria. Using these potential miRNAs sequences, we further blasted the publicly available mRNA database and detected 96 potential targets in G. hirsutum. According to the mRNA information provided by the National Center for Biotechnology Information (NCBI) (http://www.ncbi.nlm.nih.gov/), most of the miRNA targeted genes were predicted to encode transcription factors that regulate cell growth and development, signaling, and metabolism. So far, little is known about experimental or computational identification of miRNA in G. hirsutum species. These new miRNAs and their targets in G. hirsutum have been run through miRNAassist to yield data that may help us better understanding of the possible role of miRNAs in regulating the growth and development of G. hirsutum. PMID- 17408885 TI - Correlation of cellulase gene expression and cellulolytic activity throughout the gut of the termite Reticulitermes flavipes. AB - Termites have developed cellulose digestion capabilities that allow them to obtain energy and nutrition from nutritionally poor food sources, such as lignocellulosic plant material and residues derived from it (e.g., wood and humus). Lower termites, which are equipped with both endogenous (i.e., of termite origin) and symbiotic cellulases, feed primarily on wood and wood-related materials. This study investigated cellulase gene diversity, structure, and activity in the lower termite, Reticulitermes flavipes (Kollar). We initially used a metagenomics approach to identify four genes encoding one endogenous and three symbiotic cellulases, which we refer to as Cell-1, -2, -3 and -4. These four genes encode proteins that share significant sequence similarity with known endoglucanases, exoglucanases and xylanases. Phylogenetic analyses further supported these inferred relationships by showing that each of the four cellulase proteins clusters tightly with respective termite, protozoan or fungal cellulases. Gene structure studies revealed that Cell-1, -3 and -4 are intron free, while Cell-2 contains the first intron sequence to be identified from a termite symbiont cellulase. Quantitative real-time PCR (qRT-PCR) revealed that the endogenous Cell-1 gene is expressed exclusively in the salivary gland/foregut, whereas symbiotic Cell-2, -3, and -4 are highly expressed in the hindgut (where cellulolytic protists are harbored). Cellulase activity assays mapped the distribution pattern of endoglucanase, exoglucanase and xylanase activity throughout the R. flavipes digestive tract. Cellulase gene expression correlated well with the specific types of cellulolytic activities observed in each gut region (foregut+salivary gland, midgut and hindgut). These results suggest the presence of a single unified cellulose digestion system, whereby endogenous and symbiotic cellulases work sequentially and collaboratively across the entire digestive tract of R. flavipes. PMID- 17408886 TI - Direct observation of histone H2B-YFP fusion proteins and transport of their mRNA between conjugating Paramecia. AB - Cytoplasmic exchange between conjugating cells of Paramecium caudatum has been implicated by mating experiments using wild-type and behavioral mutant cells. To observe macromolecular transport between mating cells, we cloned and expressed the P. caudatum histone H2B gene as a fusion protein attached to an enhanced yellow fluorescent protein (YFP) named PcVenus. Significant fluorescent signals derived from histone H2B-PcVenus were detected throughout the macro- and micronuclei of transformant cells after microinjection of the expression vector. The normal growth and high mating reactivity of the transformants indicated that H2B-PcVenus functioned normally. Seven hours after a transformant cell expressing histone H2B-PcVenus was mated with an untransformed complementary mating-type cell, fluorescence derived from histone H2B-PcVenus was emitted from the macronuclei of the untransformed cell. About 48 h later, the fluorescent signal was detected not only in the macro- and micronuclei of untransformed cells but also in the macronuclear anlagen of both mating cells. This suggests that conjugant cells share parental histones during meiosis and subsequent DNA rearrangement. Single-cell RT-PCR analysis demonstrated the presence of H2B PcVenus mRNA in untransformed cells 15 and 24 h after conjugation. We concluded that at least the mRNA of histone H2B-PcVenus was transferred from the transformed, to the untransformed cell during conjugation. PMID- 17408887 TI - Silicateins, the major biosilica forming enzymes present in demosponges: protein analysis and phylogenetic relationship. AB - Silicateins are enzymes, which are restricted to sponges (phylum Porifera), that mediate the catalytic formation of biosilica from monomeric silicon compounds. The silicatein protein is compartmented in the sponges in the axial filaments which reside in the axial canals of the siliceous spicules. In the present study silicatein has been isolated from the freshwater sponge Lubomirskia baicalensis where it occurs in isoforms with sizes of 23 kDa, 24 kDa and 26 kDa. Since the larger protein is glycosylated we posit that it is a processed form of one of the smaller size forms. The silicatein isoforms are post-translationally modified by phosphorylation; at least four isoforms exist with pI's of 5.4, of 5.2, of 4.9 and of 4.7. Surprisingly silicatein not only mediates polymerization of silicate, but also displays proteolytic activity which is specific for cathepsin L enzymes, thus underscoring the high relationship of the silicateins to cathepsin L. The cDNAs from L. baicalensis for silicatein and cathepsin L, as well as the respective genes, were cloned. It was found that the five introns present in the sponge genes are highly conserved up to human cathepsin L. This analysis has been completed by sequencing of two silicatein genes (both for silicatein-alpha and beta) and of cathepsin L from another demosponge, Suberites domuncula. A comprehensive phylogenetic analysis with these new sequences shed new light upon the evolution of cathepsin L and silicatein families which occurred at the base of the metazoan phyla. It is concluded, that in parallel with the emergence of these enzymes at first the number of introns increased, especially in the coding region of the mature enzyme. Later in evolution the number of introns decreased again. We postulate that modification of the catalytic triad, especially of its first amino acid, is a suitable target for a chemical modulation of enzyme function of the silicateins/cathepsin L. PMID- 17408888 TI - Antiapoptotic activity of bovine herpesvirus type-1 (BHV-1) UL14 protein. AB - Viruses have evolved different strategies to interfere with apoptotic pathways in order to halt cellular responses to infection. One previous study showed that transient transfection of bovine herpesvirus type-1 (BHV-1) UL14 protein is efficient in protecting Madin Darby kidney (MDBK) and human chronic myelogenous leukemia (K562) cells from sorbitol-induced apoptosis. This protein corresponds to a putative protein of BHV-1, which shares aminoacid sequence with a part of the peptide-binding domain conserved in human heat shock protein (HSP70) family. The pBK-CMV-UL14 plasmid transfected MDBK cells treated with sorbitol did not show caspase-3 and caspase-9 activation with respect to non-transfected MDBK cells (UL14 negative). Furthermore, we report that the expression of the full length sequence of BHV-1 UL14 is evident after 7 h of infection of BHV-1 on MDBK cells which were then treated with sorbitol. These results indicate that UL14 gene product has important implications to enhance cell survival in response to apoptotic stimuli. PMID- 17408889 TI - Assessment of the variation in menopausal symptoms with age, education and working/non-working status in north-Indian sub population using menopause rating scale (MRS). AB - OBJECTIVE: To study the variation of the menopause rating scale (MRS) scores with age, working/non-working and educated/uneducated status in a cohort of north Indian subpopulation and to look for the possible reasons for the incurred variations. MRS is a well-known and validated instrument for assessing the frequency and intensity of menopausal symptoms. METHOD: A menopause clinic was organized in collaboration with a primary care centre (under the guidance of a gynecologist). A random sample of 208 women aged 35-65 years participated in the study. The MRS scale, a self-administered standardized questionnaire was applied with additional patient related information (age at menopause, level of education, working/non-working and exercising or not). RESULTS: The results were evaluated for psychological (P), somatic (S), and urogenital (U) symptoms. The average age at which menopause set in, in the cohort was found to be 48.7+/-2.3 years (46.4-51 years). Based on the average age at the menopause, the cohort was divided into peri (35-45), menopausal/early menopause (46-51) and the postmenopausal (52-65) groups. A significantly higher % of perimenopausal women (36%) showed a P score of > or =7; while a higher % of postmenopausal showed S score and U score > or =7 (>40%; p< or =0.001). Working women seem to suffer more from psychological symptoms whereas non-working women showed a greater incidence of somatic symptoms. Educated women showed a lower incidence of psychological and somatic symptoms. CONCLUSIONS: Present study indicates that age, level of education and working/non-working status (in a group of women with same socio cultural background) may also contribute to significant variations in menopausal symptoms. PMID- 17408890 TI - alpha-Cyclodextrin/oil beads: an innovative self-assembling system. AB - The aim of this work was to characterise a new type of particulate system, named beads, prepared by a straightforward technique starting from a mixture of alpha cyclodextrin aqueous solution and soybean oil without the use of any organic solvent or surface-active agent. Mechanisms involved in bead formation were also investigated. Optimal ratio between alpha-cyclodextrin (6%, w/w), soybean oil (19.6%, w/w) and water (74.4%, w/w) led to homogeneous bead size (1.6 mm) with a fabrication yield superior to 80% after a continuous external shaking during 2.5 days. After freeze-drying, oil and alpha-cyclodextrin contents were estimated at 80% (w/w) and 20% (w/w), respectively. X-ray diffraction studies revealed that beads presented a crystalline organisation and microscopic techniques showed that their inner structure was constituted by a matrix containing oily compartments. Beads offer interesting prospects for the microencapsulation of lipophilic and poorly stable molecules. Due to their semi-solid consistency and their ability to be freeze-dried, these beads have great potentialities for pharmaceutical (oral and topical routes) and cosmetic applications. PMID- 17408891 TI - Dimensional changes, gel layer evolution and drug release studies in hydrophilic matrices loaded with drugs of different solubility. AB - The objective of this investigation was to explore the effects of drug solubility on the evolution of matrix dimensions and gel layer's during drug release and investigate the relationship between these effects and the mechanism and the rate of drug release. Two hydrophilic swellable polymers Polyox (POL) and cross-linked Carbopol (CARB) were employed as carriers. Caffeine (CAF) and theophylline (THE), two drugs having similar chemical structure but different aqueous solubility, were used as model drugs. Both drug and polymer characteristics were found to influence the dimensional changes of matrices and the development of the gel layer formed around the glassy core. The dimensional expansion in CAF matrices was always more pronounced than the THE matrices. Also the CARB matrices demonstrated greater maximum expansion and lower drug release than the POL matrices, due to a smaller degree of erosion of CARB. The dimensions of CARB/CAF matrices, unlike all the other matrices studied, exhibited a biphasic increase at early times, which was attributed to the cross-linked structure of CARB and the high solubility of CAF. With both polymers, a thinner gel layer was developed in the matrices containing the less soluble THE compared to the CAF matrices. The thickness of the gel layer increased continuously with time in the CAF matrices whereas it increased initially and after reaching a maximum started to decrease in THE matrices. All formulations except those of CARB/THE exhibited burst release, which depended on drug and polymer characteristics. The gel layer thickness and erosion rate appeared to determine the rate of drug release from the CARB and POL formulations. The results clearly indicate that for these matrices gel thickness and fluctuation of gel thickness affect the release rate/h of drug proportionally. Analysis of the release kinetics indicated that CAF was released mainly through diffusion whereas, THE was released mainly through matrix erosion. PMID- 17408892 TI - Flavonoids, centaurein and centaureidin, from Bidens pilosa, stimulate IFN-gamma expression. AB - Bidens pilosa is used as an ethnical medicine for bacterial infection or immune modulation in Asia, America and Africa. Here, we employed an IFN-gamma promoter driven luciferase reporter construct and T cells to characterize immunomodulatory compounds from this plant based on a bioactivity-guided isolation principle. We found that PHA, a positive control, caused a six-fold increase in IFN-gamma promoter activity. In contrast, hot water crude extracts from Bidens pilosa and its butanol subfraction increased IFN-gamma promoter activity to two- and six fold, respectively. Finally, centaurein (EC(50)=75 microg/ml) and its aglycone, centaureidin (EC(50)=0.9 microg/ml), isolated from this butanol subfraction, augmented IFN-gamma promoter activity by approximately four-fold. Consistent with the role of centaurein or its aglycone in IFN-gamma regulation, we showed that centaurein induced the activity of NFAT and NFkappaB enhancers, located within the IFN-gamma promoter, in Jurkat cells. Overall, our results showed that centaurein regulated IFN-gamma transcription, probably via NFAT and NFkappaB in T cells. PMID- 17408893 TI - Immunomodulatory effect of bergenin and norbergenin against adjuvant-induced arthritis--a flow cytometric study. AB - Bergenin (1), a C-glycoside of 4-O-methyl gallic acid, isolated from rhizomes of Bergenia stracheyi (Saxifragaceae) and its O-demethylated derivative norbergenin (2), prepared from 1, are reported to show anti-arthritic activity through possible modulation of Th1/Th2 cytokine balance. Flow cytometric study showed that the oral administration of 1 and 2 at doses of 5, 10, 20, 40 and 80 mg/kg per oral dose inhibit the production of proinflammatory Th1 cytokines (IL-2, IFN gamma and TNF-alpha) while as potentiate anti-inflammatory Th2 cytokines (IL-4 and IL-5) in the peripheral blood of adjuvant-induced arthritic balb/c mice. This shows the potential Th1/Th2 cytokine balancing activity of 1 and 2 which is strongly correlated with their anti-arthritic activity. At similar dose levels, the effect of 2 was found to be more than that of 1. The oral LD(0) for 1 and 2 was more than 2000 mg/kg body weight of the mice. PMID- 17408894 TI - Antiulcerogenic activity of Zizyphus lotus (L.) extracts. AB - Oral administration of aqueous extracts of Zizyphus lotus root barks (50-200 mg/kg) leaves (50-200 mg/kg) and fruits (200-400 mg/kg) produced a significant (p<0.01) and dose dependent inhibition to the acute ulcer induced by HCl/ethanol solution. The methanolic (MeOH), ethyl acetate (EtOAc) and chloroformic (CHCl(3)) leaves extracts when administered orally at the dose of 200 mg/kg, exhibited a significant (p<0.01) inhibition of gastric lesions by 45%, 76% and 33%, respectively. Indeed, methanolic and ethyl acetate root barks extracts significantly reduced the gastric lesions by 47% and 41%, respectively. While the chloroformic root barks extract had no significant activity (19%). The effect of all extracts was compared with cimetidine (100 mg/kg, 62%) and omeprazole (30 mg/kg, 93%). Volume, pH and acidity of gastric juice were studied in pylorus ligated rats. Root barks (200 mg/kg, p<0.01), leaves (200 mg/kg, p<0.01) and fruits (400 mg/kg, p<0.05) aqueous extracts showed significant reduction of gastric juice secretion in pylorus ligated rats, whereas the other extracts did not show any significance. Thus, Zizyphus lotus extracts act essentially as cytoprotective agents, which support the antiulcer effect of this plant in the traditional medicine. PMID- 17408895 TI - [Principle, strengths and weaknesses of discrete choice modelling for eliciting public preferences for health care]. AB - Patients play an increasingly active role in the medical decision making process. Hence, methods for eliciting public preferences have become essential tools for enlightening the choices of health authorities. Discrete choice modelling is the most recently applied technique in the field of health. This article deals with the principle, the strengths and the weaknesses of this method. Often used in English-speaking countries, the discrete choice method has several advantages over other elicitation methods since it allows estimation of marginal utilities. Compliance rates are also higher and more information is provided. Considering these qualities, further research should be undertaken on discrete choice modelling in France. PMID- 17408896 TI - [Laboratory methods for the diagnosis of clinical forms of Lyme borreliosis]. AB - Methods used to diagnose Lyme borreliosis (LB) vary according to clinical presentations. A very good basis to clarify this nosological and clinical entity is the study published by the "European Concerted Action on Lyme Borreliosis" (EUCALB). In fact, only few studies were performed on cohorts of patients including all clinical forms of LB. For Erythema migrans, serology sensitivity is low (20% to 50%), while the sensitivity of culture or PCR reaches 50%. In early complicated forms, serology is more sensitive (70 to 90%) with the presence of concomitant IgG and IgM. Screening for antibodies in CSF is very useful for the diagnosis of neuroborreliosis. For this clinical form, culture or PCR sensitivity is disappointing (10 to 30%). In arthritis and acrodermatitis chronica atrophicans (ACA), IgG serology is 100% positive with very high titers; however IgM serology is only positive in 5 to 10% of the cases. In ACA, culture sensitivity ranges from 20 to 60% and PCR sensitivity from 60 to 90%. Specificity of antibodies, natural exposure to the etiologic agent, and cross-reactivity are critical for the final interpretation of serological assessment. Only the use of "serological profiles" allows the exploitation of detailed results (isotypes, intensity). In this approach, IgG avidity could be constructive. The western-blot is intended to confirm the specificity of antibodies found in screening methods (Elisa). PMID- 17408897 TI - [Lyme disease: basis for treatment strategy, primary preventive care and secondary preventive care]. AB - Lyme disease is the most common tick borne disease and is caused by Borrelia burgdorferi sensu lato. Ticks of the genus Ixodes are the vectors that transmit the infection to host mammals in endemic foci. Ixodes is infected by Borrelia at larval stage when it feeds on infected mammals. Man is an occasional host. The infection risk is linked to interaction between human and the natural environment. Strategies for prevention are closely related to the enzootic cycle of the Ixodes tick. Environmental measures to reduced tick density or host mammals are expensive, need to be repeated annually and cannot be applied to large areas. The primary prevention could be reduced to personal preventive measures such as reducing the amount of exposed skin and frequent checking for ticks. The risk of Lyme disease transmission after a tick bite is relatively low, and remains under 4%. The transmission rate depends on the duration of feeding. A rapide tick removal with fine tweezers or preferably special forceps and disinfection of the bite site appear to be the best technique. The absence of scientific evidence, and the risk of adverse events does not lead to recommending antimicrobial prophylaxis. Follow-up and educating the patients on the disease, clinical manifestation, and later primary prevention should be undertaken. PMID- 17408898 TI - The MRI of extraadrenal pheochromocytoma in the abdominal cavity. AB - The purpose of this study was to summarize the MR appearances of extraadrenal pheochromocytoma in the abdominal cavity and evaluate the capabilities of MRI in diagnosis of the tumor. Eleven consecutive patients with an extraadrenal pheochromocytoma in abdominal cavity who underwent preoperative 0.5 T (n=5) or 1.5 T (n=6) superconductor MRI and had a surgical resection were enrolled in the study. The MR scanning protocol included axial T(2)-weighted imaging with or without fat-suppressed sequences, axial and coronal uncontrast and contrast T(1) weighted sequences with or without fat suppression. The extraadrenal pheochromocytomas were found in retroperitoneum (n=5), the urinary bladder (n=1), the pelvis (n=1), the right prerenal area (n=1), the renal hilus (n=1), the left paramusculus psoas major (n=1) and liver (n=1). The mean maximal diameter of tumors was 55.9 mm (range 17.8-162.2 mm). The high signal intensity was seen on T(2)-weighted imaging in all tumors compared to muscle or liver, especially with fat suppression. The intratumoral septa and capsules were shown in 63.6% and 72.7% of cases, respectively, which had low signal intensity on T(2)-weighted imaging. These relative characteristics may be helpful for qualitative diagnosis of extraadrenal pheochromocytomas with MRI. Other usefulness of MRI was to locate the position, to decide the range of tumors and to show well the relationship between the tumor and near structures. PMID- 17408899 TI - Can independent coronal multiplanar reformatted images obtained using state-of the-art MDCT scanners be used for primary interpretation of MDCT of the abdomen and pelvis? A feasibility study. AB - PURPOSE: To evaluate if coronal reformatted images can be used for primary interpretation of MDCT of the abdomen and pelvis using 64-slice MDCT. MATERIALS AND METHODS: IRB approval was obtained. We reviewed MDCT studies of the abdomen and pelvis of 220 consecutive patients performed with 64 row MDCT with constant scanning parameters. Based on a 0.625mm raw data set, transverse images were reconstructed at 5mm and coronal images at 3mm using standard reconstruction algorithms. Reader familiarity was achieved by simultaneous evaluation of transverse and coronal reformats in an initial group of 20 separate cases for findings in consensus. Two subsequent phases of image analysis were then performed in two groups of 100 patients each. In the first phase two radiologists evaluated the added utility of simultaneous review of MDCT of transverse and coronal reformatted images over transverse images alone in 100 consecutive patients referred for MDCT of the abdomen and pelvis. In the second phase, the same radiologists evaluated whether coronal multiplanar reformats could be used for primary interpretation of MDCT of the abdomen and pelvis in a separate but similar cohort of 100 consecutive abdominopelvic MDCT studies. The number of lesion(s), their location, size of smallest lesion, presence of artifacts and likely diagnosis were noted at each image interpretation. Image quality and confidence for interpretation was evaluated using five-point and three-point scale, respectively. The time required for primary interpretation of coronal reformats and transverse images were recorded. Statistical analysis was performed using Wilcoxon signed rank test. RESULTS: Both readers detected additional findings (n=37, 35), respectively, on simultaneous review of transverse and coronal reformats as compared with transverse images alone (p<0.001). Excellent interobserver agreement was noted (r=0.94-0.96). Both readers detected additional findings (n=62, 53), respectively, on independent review of coronal reformats as compared with transverse images alone (p<0.001). Readers' confidence was also found to be higher on coronal evaluations as compared to axial images (p<0.01). There was good interobserver agreement between the two readers. CONCLUSION: Independent coronal multiplanar reformatted images obtained using state-of-the art MDCT scanners show promise as the preferred orientation and can be useful for primary interpretation of MDCT of the abdomen and pelvis. PMID- 17408900 TI - Evaluation of small (99.9% susceptible), H. influenzae (< or =0.03 microg/mL, >99.9%), and M. catarrhalis (< or =0.03 microg/mL, 100.0%). The garenoxacin potency versus the pneumococci was 16- to 32-fold greater than levofloxacin or ciprofloxacin and 2 fold superior to moxifloxacin (MIC(90), 0.12 microg/mL). Resistances to other classes of antimicrobials did not adversely influence garenoxacin MIC results. Ciprofloxacin- or levofloxacin-resistant (MIC, > or =4 microg/mL) S. pneumoniae had higher garenoxacin MIC(90) values (1 microg/mL), but 90.6% to 97.5% of strains remained susceptible. Strains of all 3 monitored pathogens with mutations in the quinolone resistance determining region (QRDR) had higher garenoxacin MIC results, with > or =3 to 4 QRDR mutations required to elevate garenoxacin MIC values to > or =2 microg/mL. In conclusion, garenoxacin possesses a potent activity against pneumococci, H. influenzae, and M. catarrhalis strains worldwide, at a level significantly greater than the available tested agents in the fluoroquinolone class (ciprofloxacin, levofloxacin, and moxifloxacin). Only 13 and 4 isolates (0.07% and 0.03%) of S. pneumoniae and H. influenzae, respectively, had a garenoxacin MIC at > or =2 microg/mL, thus, making this new "respiratory antipneumococcal" quinolone an attractive candidate for the therapy of contemporary CA-RTI (bronchitis, pneumonia, and sinusitis). PMID- 17408904 TI - Garenoxacin activity against isolates form patients hospitalized with community acquired pneumonia and multidrug-resistant Streptococcus pneumoniae. AB - Community-acquired pneumonia (CAP) continues to cause significant morbidity worldwide, and the principal bacterial pathogens (Streptococcus pneumoniae and Haemophilus influenzae) have acquired numerous resistance mechanisms over the last few decades. CAP treatment guidelines have suggested the use of broader spectrum agents, such as antipneumococcal fluoroquinolones as the therapy for at risk patient population. In this report, we studied 3087 CAP isolates from the SENTRY Antimicrobial Surveillance Program (1999-2005) worldwide and all respiratory tract infection (RTI) isolate population of pneumococci (14665 strains) grouped by antibiogram patterns against a new des-F(6)-quinolone, garenoxacin. Results indicated that garenoxacin was highly active against CAP isolates of S. pneumoniae (MIC(90), 0.06 microg/mL) and H. influenzae (MIC(90), < or =0.03 microg/mL). This garenoxacin potency was 8- to 32-fold greater than gatifloxacin, levofloxacin, and ciprofloxacin against the pneumococci and >99.9% of strains were inhibited at < or =1 microg/mL (proposed susceptible breakpoint). Garenoxacin MIC values were not affected by resistances among S. pneumoniae strains to penicillin or erythromycin; however, coresistances were high among the beta-lactams (penicillins and cephalosporins), macrolides, tetracyclines, and trimethoprim/sulfamethoxazole. Analysis of S. pneumoniae isolates with various antimicrobial resistance patterns to 6 drug classes demonstrated that garenoxacin was active against >99.9% (MIC, < or =1 microg/mL) of strains, and the most resistant pneumococci (6-drug resistance, 1051 strains or 7.2% of all isolates) were completely susceptible (100.0% at < or =1 microg/mL) to garenoxacin (MIC(90), 0.06 microg/mL). These results illustrate the high activity of garenoxacin against contemporary CAP isolates and especially against multidrug resistant (MDR) S. pneumoniae that have created therapeutic dilemmas for all RTI presentations. Garenoxacin appears to be a welcome addition to the CAP treatment options, particularly for the emerging MDR pneumococci strains. PMID- 17408905 TI - Demyelinating diseases and potential repair strategies. AB - Demyelination is associated with a number of neurological disorders including multiple sclerosis (MS), spinal cord injury and nerve compression. MS lesions often show axon loss and therefore reparative therapeutic goals include remyelination and neuroprotection of vulnerable axons. Experimental cellular transplantation has proven successful in a number of demyelination and injury models to remyelinate and improve functional outcome. Here we discuss the remyelination and neuroprotective potential of several myelin-forming cells types and their behavior in different demyelination and injury models. Better understanding of these models and current cell-based strategies for remyelination and neuroprotection offer exciting opportunities to develop strategies for clinical studies. PMID- 17408906 TI - Influence of thyroid hormones on maturation of rat cerebellar astrocytes. AB - Thyroid hormone influences brain maturation through interaction with nuclear receptors and regulation of gene expression. Their role on astrocyte maturation remains unclear. We have analyzed the role of thyroid hormone in rat cerebellar astrocyte maturation by comparing the sequential patterns of intermediate filament expression in normal and hypothyroid animals. During normal development astroglial cells sequentially express nestin, vimentin, and glial fibrillary acidic protein. Differentiated astrocytes appeared in the superior medullary vellum by postnatal day 2 and reached the white mater and internal granular layer by postnatal day 4. Intermediate filament marker expression was transiently lost from postnatal days 6 to 8 in anterior lobes, without an increased apoptosis. Vimentin expression was replaced by glial fibrillary acidic protein between postnatal days 10 and 32. The differentiated astrocytes were evenly distributed throughout the cerebellar slices, including the internal granular layer. Differences between normal and hypothyroid rats were observed starting from postnatal day 4, with lack of differentiated astrocytes in the internal granular layer. The transient decrease of astrocyte markers immunoreactivity in the anterior lobe did not take place in hypothyroid rats. The vimentin-glial fibrillary acidic protein transition was delayed and most differentiated astrocytes remained confined to the white matter. The results indicate that thyroid hormone deficiency induces a delay and a partial arrest of astrocyte differentiation. Astrocytes express thyroid hormone receptor alpha and beta subtypes suggesting that astrocytes are direct target cells of thyroid hormones. PMID- 17408907 TI - Prudent application of radiofrequency ablation in resectable colorectal liver metastasis. AB - Radiofrequency ablation (RFA) for liver metastasis of colorectal (H-CRC) origin is a well-documented technique in surgically unresectable disease. Overall recurrence figures appear inferior to resection but are based on a selection of patients with unresectable disease, often due to multiple localisations of extensive disease. Lesion based recurrence is probably more appropriate to predict results of RFA in surgically resectable H-CRC and figures may be good enough to consider RFA an alternative treatment in high risk patients. PMID- 17408908 TI - Chk1 frameshift mutation in sporadic and hereditary non-polyposis colorectal cancers with microsatellite instability. AB - AIM: Protein kinase Chk1 (hChk1) is essential in human cells for cell cycle arrest in response to DNA damage, and has been shown to play an important role in the G2/M checkpoint. The BRAF mutations have been suggested to be linked with defective mismatch repair in colorectal cancers. The aim of this study was to investigate whether a frameshift mutation within the Chk1 gene contribute to the development or progression of eastern sporadic and hereditary non-polyposis colorectal cancer (HNPCC) with microsatellite instability (MSI). METHODS: We analyzed MSI using the 6 microsatellite markers and a frameshift mutation in the BRAF gene and in poly(A)9 within the Chk1 gene in 51 sporadic colorectal cancer and 14 HNPCC specimens. RESULTS: Eleven of the 51 sporadic colorectal cancers and all of the 14 HNPCCs were MSI-positive. Chk1 frameshift mutations were observed in 2 and 3 sporadic colon cancers and HNPCC, respectively, whereas no BRAF mutations were detected in these samples. Interestingly, all cases with the Chk1 frameshift mutation had high-frequency MSI. CONCLUSION: These results suggest that the Chk1 gene is a target of genomic instability in MSI-positive colorectal cancers and that the Chk1 framshift mutations might be involved in colorectal tumourigenesis through a defect in response to DNA damage in a subset of sporadic colorectal cancers and HNPCCs. PMID- 17408909 TI - Evaluation of the risk of contamination of surgical personnel by vaporization of oxaliplatin during the intraoperative hyperthermic intraperitoneal chemotherapy (HIPEC). AB - AIMS: To evaluate if oxaliplatin is likely to vaporize under HIPEC conditions and to see if it could be a source of pulmonary contamination for surgeons. METHODS: Three oxaliplatin concentrations (230, 460 and 920 mg oxaliplatin/l), 3 heating temperatures (41, 43 and 45 degrees C) and 3 bubbling durations (30, 60 and 90 min) were tested. Drug vaporization was evaluated by using inductively coupled plasma mass spectrometry (ICP-MS) to analyze platinum concentrations in the trap solutions. RESULTS: At all concentrations of oxaliplatin solutions, heating temperatures and bubbling trap periods, the quantities of vaporized platinum were always insignificantly lower than 1 microg/l. CONCLUSIONS: The experimental risk of pulmonary contamination of hospital staff during HIPEC procedure appears to be negligible. However a monitoring study with an analysis of samples of the operating theatre and urine from surgical personnel should be carried out to confirm these conclusions. PMID- 17408910 TI - Prostate cancer screening strategies with re-screening interval determined by individual baseline prostate-specific antigen values are cost-effective. AB - AIMS: To determine whether prostate cancer screening strategies with re-screening interval determined by individual baseline prostate-specific antigen values are cost-effective. METHODS: Based on the results of an actual contemporary screening program, we established Markov decision analytic models of prostate cancer screening with personalized re-screening interval strategies using cutoff baseline PSA levels for biennial screening as well as a model of uniformly annual or biennial screening. These strategies were compared in terms of cumulative incidence of early cancer and cost-effectiveness. RESULTS: Early cancer detection rates were similar among all strategies. Personalized strategies were more cost effective compared to uniform screening strategies. If all participants with negative PSA results uniformly omit annual screening, it would be more costly but less effective (dominated). Contrary, annual screening for all participants would cost too much. These results were robust throughout sensitivity analysis incorporating every assumption in the models. CONCLUSIONS: This study adds important evidence that personalized rescreening strategies based on individual baseline PSA have advantages of cost-effectiveness against conventional uniform strategies. PMID- 17408911 TI - [Selection of medical students: a process influenced by history and culture]. AB - OBJECTIVES: Every medical school in the world has a procedure for selecting students for medical studies, intended to identify those who are capable of becoming doctors, according to specific expectations. The means of selection, however, vary significantly from one school to another and from one country to another. France has chosen to undertake this selection at the end of the first year of university studies. This method is costly in that it requires medical schools to organize a whole year of curriculum for thousands of students, fewer than 30% of whom will be chosen for further medical studies. It is also counterproductive for students, a significant proportion of whom are not admitted despite above-average results on their examinations. METHODS: This article examines the historical and cultural factors that have led to the current organization. Our study is based on an analysis of government archives dealing with medical education reforms from 1905-1970. RESULTS: We argue that the format of the current selection process is linked to ideas of freedom of access for students and of equal opportunity for admission to medical studies. CONCLUSION: We conclude that for any reform to be acceptable it must take into account the historical and cultural elements (for the most part stemming from the French Revolution) that form the basis of the current system. PMID- 17408912 TI - [Treatment of ANCA-associated vascularitides]. AB - Treatment for ANCA-associated vasculitides is now well defined, but must be adjusted for each patient according to the type of vasculitis, its precise form (e.g., limited versus systemic Wegener's granulomatosis) and severity, and patients' characteristics, such as age and renal function. The therapeutic decision must also take into account the risk of adverse events inherent to each treatment. The efficacy of adequate induction treatment has been demonstrated: more than 80% of patients now achieve remission. Relapse rates nonetheless remain high, especially in Wegener's granulomatosis. Patients with microscopic polyangiitis or Churg-Strauss syndrome with no poor prognostic factors can be treated with corticosteroids alone, with immunosuppressants added only in case of treatment failure. Patients with Wegener's granulomatosis or microscopic polyangiitis or Churg-Strauss syndrome and one or more poor prognostic factors must receive a combination of corticosteroids and immunosuppressants, mainly intravenous pulsed cyclophosphamide. Plasma exchange is indicated as an adjuvant therapy for patients with severe renal involvement. Once remission is achieved, maintenance therapy can replace cyclophosphamide by a less toxic immunosuppressive drug, such as azathioprine or methotrexate. For these latter patients, the optimal duration of induction therapy remains to be determined, but should not be shorter than 18 months. Conversely, there is no need to prescribe high-dose corticosteroids for months. Prednisone must be started at 1 mg/kg/d then rapidly tapered so that patients are not receiving more than 15 mg/d after 3 4 months of therapy. Biological therapies also appear to have a place in the therapeutic armamentarium for ANCA-associated systemic vasculitides, at least for patients whose disease is refractory to conventional therapy. However, the precise indications for anti-TNFalpha or anti-CD20 monoclonal antibodies and their optimal regimens (doses and durations) have not yet been defined. Anti-IL5, interferon-alpha and anti-IgE monoclonal antibodies might also be useful for Churg-Strauss syndrome. These biologics must be prescribed extremely cautiously and only in trial settings, especially in view of the adverse effects, few but severe, recently been reported with them. PMID- 17408913 TI - [Improving patient safety: decreasing adverse events associated with medical care]. AB - Awareness of the importance of what were previously called iatrogenic accidents is not new, but recent publications have demonstrated the frequency and severity of the accidents and incidents associated with care, which are now known simply as "adverse events". Research has helped us to understand the principal mechanisms underlying them and the circumstances that promote them. It shows that root causes, often linked to the organization of care, should be sought beneath the initial appearance of mistakes. Institutions providing health care must ascertain how to develop a new culture that makes it possible to improve patient safety by implementing new policies, that is, a group of several coordinated measures intended to decrease patient risk. These policies should use accepted techniques, such as reports and appropriate information management for events for which reporting is mandatory, but extended to medical accidents; critical activity analyses must also be used, for comparison with a standard, following the model used for evaluations of professional practices. New techniques are also necessary, such as operational feedback in the form of morbidity-mortality reviews and in-depth analyses of the most serious events. Institutions must establish indicators to prove the effectiveness of this new policy. PMID- 17408914 TI - [Pulmonary imaging in ANCA-associated vasculitides]. AB - Imaging of ANCA-associated vasculitides principally shows nonsystematized alveolar opacities, predominantly central, that suggest alveolar hemorrhage, as well as uni- or multifocal alveolar opacities of variable interpretation, and nodules, including cavitary nodules. Other signs, observed more rarely on imaging, are interstitial lung diseases, tracheobronchial involvement, and pulmonary hypertension. The principal pulmonary signs of Wegener's granulomatosis are nodules and masses, sometimes cavitary, and areas of airspace consolidation that may or may not suggest diffuse alveolar hemorrhage. Except when the latter is present, the lesions correspond to necrotizing granulomatosis or organized pneumonia. The most frequent pulmonary signs of Churg-Strauss syndrome on computed tomography are ground glass areas or parenchymal consolidation in dispersed bands or with a predominantly subpleural distribution that expresses eosinophilic interstitial and alveolar infiltration. Alveolar hemorrhage is the most common expression of microscopic polyangiitis. PMID- 17408915 TI - [Classification of systemic vasculatides]. AB - Vasculitides are defined by inflammation of blood vessel walls leading to vascular stenosis or occlusion, with various degrees of fibrinoid necrosis of the media and inflammatory infiltration, mainly neutrophilic and sometimes granulomatous. Various classifications of the vasculitides have been proposed. The classifications used most today are the 1990 American College of Rheumatology classification and the Chapel Hill nomenclature, published in 1994 . Only the latter distinguished between polyarteritis nodosa and microscopic polyangiitis and stressed the importance of antineutrophil cytoplasm autoantibodies (ANCA). In practice, primary systemic vasculitides are classified according to their clinical presentations, their precise histological features, and the size of the predominantly affected vessels. Some small-vessel vasculitides are associated with the presence of ANCA: 90% of patients with systemic Wegener's granulomatosis (mainly ANCA with cytoplasm labeling on indirect immunofluorescence and proteinase 3 specificity), 80% of the subjects with microscopic polyangiitis (mostly pANCA with myeloperoxidase specificity), and more than one third of those with Churg-Strauss syndrome (mostly pANCA). PMID- 17408916 TI - [Churg-Strauss syndrome]. AB - Churg-Strauss syndrome is a systemic and pulmonary vasculitis, defined by its association with severe asthma and with hypereosinophilia of the blood and tissues. The systemic vasculitis is a small-vessel vasculitis frequently associated with purpura, mononeuritis multiplex, and, more rarely, with rapidly progressive glomerulonephritis or diffuse alveolar hemorrhage. Its prevalence of 7 to 13 per million population makes it one of the rarest of the systemic vasculitides. Anti-MPO (antimyeloperoxidase) pANCA (ANCA with a perinuclear fluorescence pattern) is present in 35-40% of cases and appears to determine a subgroup of patients with a higher frequency of renal damage, alveolar hemorrhage, and central nervous system damage. Cardiac involvement is an important cause of morbidity and the leading cause of mortality in Churg-Strauss syndrome. Treatment is based on corticosteroid therapy and immunosuppressive drugs (cyclophosphamide and azathioprine) and is determined according to validated prognostic criteria (Five-Factor Score). Complete remission occurs in almost 90% of cases, and the 10-year survival rate has reached 79.4%. Relapses are frequent (25% of cases) and even after recovery from vasculitis, most patients (90%) still have asthma requiring corticosteroid treatment. PMID- 17408917 TI - [Nasogastric tube feeding in bulimia. Controlled study with follow-up at 3 months]. AB - OBJECTIVE: Few effective treatments are available for severe forms of bulimia nervosa, which are accompanied by malnutrition, anxiety, and depressive mood. We previously showed in an open study that nasogastric tube feeding (TF) reduced binges and purging in patients with anorexia nervosa. METHODS: This prospective randomized trial compared bulimia patients in two treatment groups: one group received TF at home, together with psychotherapy, nutritional counseling and a support group while the control group received only psychotherapy, nutritional counseling, and a support group. Patients in the first group underwent TF for 8 weeks (exclusively for 10 days and associated with meals thereafter). Assessment was based on clinical examination, laboratory results, and a variety of questionnaires (our in-house instrument for measuring binge and vomiting episodes, eating disorder inventory, Beck's depression inventory and the Hamilton rating scale for anxiety), all performed at the onset of treatment and at 8 days, 8 weeks (i.e., the end of TF), and 3 months after treatment began. RESULTS: Binges and vomiting disappeared faster and more frequently in TF patients than in the control group: 65% versus 29% (p<0.01). Three months later, these remained less frequent in the TF group than among controls (52% versus 33%, p=0.064). Nutritional status, depression, and anxiety improved more among the TF than control subjects (p<0.05). CONCLUSION: Tube feeding was effective in these patients with bulimia nervosa, reducing the number of binge and vomiting episodes and improving nutritional status and mood. PMID- 17408918 TI - Effects of benzo(e)pyrene and benzo(a)pyrene on P-glycoprotein-mediated transport in Caco-2 cell monolayer: a comparative approach. AB - The previous studies from our laboratory reported that benzo(a)pyrene (Bap) influenced efflux transport of rhodamine 123 (Rho-123) by induction of P glycoprotein (P-gp) in Caco-2 cells. The present study investigated whether induction of P-gp and the enhanced efflux transport of Rho-123 were caused by benzo(e)pyrene (Bep), which has a structure similar to Bap, but is not a carcinogenic compound. In Caco-2 monolayer exposed to 50 microM Bep for 72 h, the ratio of the apparent permeability coefficient (P(app)) of Rho-123 efflux increased significantly compared to that of the control monolayer. Similarly, a significant increase in expression of MDR1 mRNA and of P-gp at the protein level were detected by RT-PCR and by Western blot analysis, respectively, in Caco-2 cells exposed to Bep, compared to that of the control. Caco-2 cells exposed to Bep showed oxidative stress that was detected by fluorescence microscopy using aminophenyl fluorescein. However, the oxidative stress was weaker compared with that of Bap. The cellular GSH content was decreased to 80% or 59% of control cells, respectively, in Caco-2 cells exposed to either Bep or Bap. Our results further show that Bep or Bap-induced P-gp in Caco-2 cells might have been the result of oxidative stress rather than DNA damage. PMID- 17408920 TI - The two-component signal system in rice (Oryza sativa L.): a genome-wide study of cytokinin signal perception and transduction. AB - In this report we define the genes of two-component regulatory systems in rice through a comprehensive computational analysis of rice (Oryza sativa L.) genome sequence databases. Thirty-seven genes were identified, including 5 HKs (cytokinin-response histidine protein kinase) (OsHK1-4, OsHKL1), 5 HPs (histidine phosphotransfer proteins) (OsHP1-5), 15 type-A RRs (response regulators) (OsRR1 15), 7 type B RR genes (OsRR16-22), and 5 predicted pseudo-response regulators (OsPRR1-5). Protein motif organization, gene structure, phylogenetic analysis, chromosomal location, and comparative analysis between rice, maize, and Arabidopsis are described. Full-length cDNA clones of each gene were isolated from rice. Heterologous expression of each of the OsHKs in yeast mutants conferred histidine kinase function in a cytokinin-dependent manner. Nonconserved regions of individual cDNAs were used as probes in expression profiling experiments. This work provides a foundation for future functional dissection of the rice cytokinin two-component signaling pathway. PMID- 17408921 TI - Methylmercury toxicity and functional programming. AB - PURPOSE: Adverse health effects of developmental toxicants may induce abnormal functional programming that leads to lasting functional deficits. This notion is considered from epidemiological evidence using developmental methylmercury neurotoxicity as an example. MOST IMPORTANT FINDINGS: Accumulating evidence indicates that adverse effects may occur even at low-level methylmercury exposures from seafood and freshwater fish. Neurobehavioral outcomes are usually non-specific, and imprecise exposure assessment results in a bias toward the null. Essential nutrients may promote the development of certain brain functions, thereby causing confounding bias. The functional deficits caused by prenatal methylmercury exposure appear to be permanent, and their extent may depend on the joint effect of toxicants and nutrients. PRINCIPAL CONCLUSIONS: The lasting functional changes caused by neurodevelopmental methylmercury toxicity fit into the pattern of functional programming, with effects opposite to those linked to beneficial stimuli. PMID- 17408922 TI - Predictor factors for childhood obesity in a Spanish case-control study. AB - OBJECTIVE: The aim of this study was to identify and characterize factors associated with childhood obesity risk related to lifestyle and perinatal life influences (dietary pattern, physical activity, family history of obesity, breast feeding, sedentary behavior, and birth weight) in a case-control study design. METHODS: Cases were 185 obese (body mass index >97th percentile) children and adolescents (6-18 y old) from the Navarra region of Spain and control subjects were matched by sex and age. Anthropometric data were collected, and a personal interview about lifestyle parameters (dietary patterns, physical activity, family history of obesity, breast-feeding and sedentary behaviors) was performed with each participant. Conditional logistic regression was used to identify predictive factors of obesity. RESULTS: Leisure time physical activity proved to be a protector factor against obesity (odds ratio [OR] 0.94, 95% confidence interval [CI] 0.92-0.96), whereas family history of obesity (OR 4.18, 95% CI 2.20-8.62), watching television (OR 2.02, 95% CI 1.09-3.77), and sugar-sweetened beverage consumption (OR 1.74, 95% CI 1.05-2.89) were associated with a higher obesity risk. CONCLUSION: Our results suggest that leisure time physical activity, family history of obesity, watching television, and sugar-sweetened beverage consumption are important predictive variables for childhood obesity, whereas other factors including breast-feeding, birth weight, and time spent sleeping did not appear to play an important role in the development of childhood obesity. PMID- 17408923 TI - Prolonged survival with confirmed metastatic pulmonary ameloblastoma. AB - This report presents a living 94-year-old female with known metastatic pulmonary ameloblastoma diagnosed over 37 years ago. Initial treatment for the primary lesion of the anterior mandible was carried out 18 years before diagnosis of pulmonary lesions. Despite the presence of bilateral pulmonary metastases, the patient remains asymptomatic and received no treatment for these lesions. To our knowledge this case represents the longest survival time recorded after appearance of untreated metastatic disease. PMID- 17408924 TI - Single-dose and multi-dose clindamycin therapy fails to demonstrate efficacy in preventing infectious and inflammatory complications in third molar surgery. AB - The goal of this study was to evaluate the efficacy of single- and multi-dose (5 day) clindamycin therapy for the prevention of inflammatory complications in patients undergoing lower third molar surgical extraction with bone removal. Patients who qualified for the prospective, randomized, double-masked, placebo controlled trial were randomly divided into three groups: (1) single dose of oral clindamycin administered preoperatively (single-dose group); (2) clindamycin administered preoperatively with continued therapy for 5 days (5-day group); and (3) a placebo group. The following parameters were evaluated on the first, second and seventh days postsurgery: trismus, facial swelling, body temperature, lymphadenopathy, alveolar osteitis and subjective pain sensations. There were 86 patients (31 in the single-dose group, 28 in the 5-day group and 27 in the placebo group) enrolled in the study. There were no statistically significant differences in postoperative inflammatory complications in patients during the first and second days postsurgery. A statistically significant variation in body temperature was reported on the seventh day. Analysis of the postoperative analgesic intake did not show statistically significant differences between examined groups. Clindamycin applied in a single preoperative dose of 600 mg with or without subsequent 5-day therapy does not demonstrate efficacy in prophylaxis for postoperative inflammatory complications after third molar surgery. PMID- 17408925 TI - Alterations in L-arginine-nitric oxide-producing pathway affect antioxidative defense in the rat skin. PMID- 17408926 TI - Quorum-sensing-dependent switch to butanediol fermentation prevents lethal medium acidification in Aeromonas hydrophila AH-1N. AB - Aeromonas hydrophila AH-1N shows biphasic growth in glucose containing broth at 30 degrees C. The first phase is characterized by strong acidification, while the second phase is accompanied by an increase in medium pH. Disruption of AHL production by knockout of ahyI did not affect the first growth phase, but resulted in further acidification which completely blocked the second growth phase and even caused cell death. The block could be relieved by addition of 5 microM (or less) synthetic N-butanoyl-L-homoserine lactone (C4-HSL), the major AHL produced by A. hydrophila AH-1N. Further analysis revealed that, as shown previously for Serratia plymuthica, butanediol fermentation is regulated by AHL mediated quorum sensing in A. hydrophila AH-1N. These results indicate that A. hydrophila switches to butanediol fermentation to reduce medium acidification under certain conditions and to allow further growth if nutrient resources are not yet exhausted. PMID- 17408927 TI - Genetic and proteomic characterization of rifaximin resistance in Bifidobacterium infantis BI07. AB - Rifaximin resistance in the probiotic strain Bifidobacterium infantis BI07 was studied to assess the use of an antibiotic-probiotic combination for clinical management of intestinal disorders. A rifaximin-resistant mutant was selected and a 129 bp core region of the rpoB gene was sequenced and compared with the respective sequence of the sensitive clone. A miss-sense mutation of codon 513, producing the substitution of Gln with Arg in the protein sequence, was found. The involvement of metabolic changes associated with rifaximin resistance was also investigated by proteomic analysis performed with two-dimensional electrophoresis and mass spectrometry. The principal categories of proteins, whose expression levels varied as a consequence of rifaximin resistance, included chaperonins, regulatory factors and metabolic enzymes. The hypothesis of rifaximin inactivation by bacterial enzymatic activities was excluded, as neither structural modifications nor degradation derivates of the drug moiety was identified using liquid chromatography coupled with tandem mass spectrometry. PMID- 17408928 TI - A blood isolate of Neisseria meningitidis showing reduced susceptibility to quinolones in Hong Kong. PMID- 17408929 TI - Update on human prion disease. AB - The recognition that variant Creutzfeldt-Jakob disease (vCJD) is caused by the same prion strain as bovine spongiform encephalopathy in cattle has dramatically highlighted the need for a precise understanding of the molecular biology of human prion diseases. Detailed clinical, pathological and molecular data from a large number of human prion disease patients indicate that phenotypic diversity in human prion disease relates in part to the propagation of disease-related PrP isoforms with distinct physicochemical properties. Incubation periods of prion infection in humans can exceed 50 years and therefore it will be some years before the extent of any human vCJD epidemic can be predicted with confidence. PMID- 17408930 TI - Diffusion tensor imaging in dementia with Lewy bodies and Alzheimer's disease. AB - Dementia with Lewy bodies (DLB) is a common form of dementia, with fewer memory deficits, and more visuo-perceptual problems than Alzheimer's disease (AD). We hypothesized that there would be disease specific alterations revealed by diffusion tensor imaging with AD showing temporal lobe and DLB more parietal changes. We recruited 15 people with AD, 16 with DLB, and 15 healthy control subjects of similar age. They were scanned on a 1.5 T MRI system with diffusion tensor FLAIR imaging. Apparent diffusion coefficient (ADC) and fractional anisotropy (FA) maps were calculated, and data were analysed using pre-defined regions of interest (ROI) and also with SPM. We found a significant decrease in the FA map in a ROI in the parietal lobe (precuneus) of the DLB group. Using SPM we found increased ADC in the left temporal lobe of AD subjects compared to controls. There were no other significant differences between groups. We conclude that there are subtle changes visible with diffusion imaging in DLB and AD which may reflect disrupted connectivity and underlie observed perfusion changes in these disorders. PMID- 17408931 TI - Protein microarrays and quantum dot probes for early cancer detection. AB - We describe here a novel approach for detection of cancer markers using quantum dot protein microarrays. Both relatively new technologies; quantum dots and protein microarrays, offer very unique features that together allow detection of cancer markers in biological specimens (serum, plasma, body fluids) at pg/ml concentration. Quantum dots offer remarkable photostability and brightness. They do not exhibit photobleaching common to organic fluorophores. Moreover, the high emission amplitude for QDs results in a marked improvement in the signal to noise ratio of the final image. Protein microarrays allow highly parallel quantitation of specific proteins in a rapid, low-cost and low sample volume format. Furthermore the multiplexed assay enables detection of many proteins at once in one sample, making it a powerful tool for biomarker analysis and early cancer diagnostics. In a series of multiplexing experiments we investigated ability of the platform to detect six different cytokines in protein solution. We were able to detect TNF-alpha, IL-8, IL-6, MIP-1beta, IL-13 and IL-1beta down to picomolar concentration, demonstrating high sensitivity of the investigated detection system. We have also constructed and investigated two different models of quantum dot probes. One by conjugation of nanocrystals to antibody specific to the selected marker--IL-10, and the second by use of streptavidin coated quantum dots and biotinylated detector antibody. Comparison of those two models showed better performance of streptavidin QD-biotinylated detector antibody model. Data quantitated using custom designed computer program (CDAS) show that proposed methodology allows monitoring of changes in biomarker concentration in physiological range. PMID- 17408932 TI - Chitosan nanoparticles are compatible with respiratory epithelial cells in vitro. AB - The aim of this work was to evaluate the biocompatibility of novel respirable powder formulations of nanoparticles (NP) entrapped in mannitol microspheres using human respiratory epithelial cell lines. Microspheres formulated at NP:mannitol ratios of 10:90, 20:80 and 40:60 were evaluated using the Calu-3 and A549 cell lines. The MTT cell viability assay revealed an absence of overt toxicity to Calu-3 or A549 cells following exposure to the formulations containing <1.3mg NP/ml (equivalent to 0.87 mg NP/cm(2)) for up to 48 h. Transepithelial electrical resistance (TER) and solute permeability in Calu-3 cell layers were determined following exposure of the cells to the NP:mannitol 20:80 formulation. After administration of the formulation dissolved in serum free cell culture medium (1.3mg/ml NP suspension) to the cells, neither TER nor permeability were altered compared to untreated cell layers. Confocal microscopy did not reveal any NP internalisation under the conditions used in this study, although evidence of mucoadhesion was observed. All the data presented are encouraging with respect to the development of chitosan NP-containing microspheres for the pulmonary administration of therapeutic macromolecules. Not only do the formulations possess suitable aerodynamic characteristics and the capacity to encapsulate proteins as shown previously; they have now been shown to exhibit in vitro biocompatibility. PMID- 17408933 TI - Effective integration of systems biology, biomarkers, biosimulation and modelling in streamlining drug development. AB - The European Federation of Pharmaceutical Sciences (EUFEPS) has long established itself as leaders in the field of interdisciplinary meetings to discuss issues that face drug development. It's ever popular and well attended "Optimizing Drug Development" series has tackled numerous issues, most recent of which have been drug interactions, getting the dose right, candidate selection, and biomarkers (Lesko et al., 2000; Rolan et al., 2003; Stanski et al., 2005; Tucker et al., 2001). Over a course of 3 productive days, the meeting on "Effective Integration of Systems Biology, Biomarkers, Biosimulation and Modelling in Streamlining Drug Development", held in Basel, Switzerland was jointly sponsored by EUFEPS, European Biosimulation Network of Excellence (BioSim), American College of Clinical Pharmacology (ACCP), European Centre of Pharmaceutical Medicine (ECPM), and Swiss Society of Pharmaceutical Sciences (SGRW). The meeting was focused on emerging aspects related to the quantitative understanding of underlying pathways in drug discovery and clinical development, i.e. moving from an empirical to a model-based, quantitative drug development process. The objectives of the meeting were: (1) to highlight the current state of the art on biomarkers (as they relate to quantitative fingerprinting of disease), systems biology, modelling and simulation; (2) to illustrate the applications of these emerging tools in increasing the efficiency and productivity of new drug development by case examples; (3) to understand the gaps in the technology and organizational implementations in governance, and (4) allow an opportunity for cross disciplinary interaction, i.e., scientists with more theoretical and technical modelling and simulation expertise of the BioSim network and researchers experienced in applying modelling and simulation techniques in day-to-day drug development were drawn together. This report summarizes the outcome from this meeting. PMID- 17408934 TI - Methotrexate and erythro-9-(2-hydroxynon-3-yl) adenine therapy for rat adjuvant arthritis and the effect of methotrexate on in vivo purine metabolism. AB - The objectives were: (1) to test the association of methotrexate (MTX) efficacy in rat adjuvant arthritis (rat AA) with interference of purine biosynthesis and adenosine metabolism and (2) to test the efficacy of erythro-9-(2-hydroxynon-3 yl) adenine (EHNA), an inhibitor of adenosine deaminase, and the efficacy of aminoimidazolecarboxamide (AICA) riboside plus MTX in rat AA. Radiographic and histologic examinations of the hind limbs were measures of efficacy. Urinary excretions of AICA and adenosine were markers of AICA ribotide transformylase inhibition (i.e., blockage of purine biosynthesis) and interference with adenosine metabolism, respectively. AICA and adenosine excretions increased during the day of MTX dosing (treatment day) compared to the previous baseline day in animals responding well to MTX (i.e., low radiographic and histologic scores). Based on radiographic and histologic scores, adjuvant injected rats were separated into two disease categories (i.e., no/mild and moderate/severe). Only AICA excretion was significantly elevated on the treatment day in rat AA with no/mild disease (i.e., those responding well to MTX therapy). AICA (not adenosine) excretion was significantly correlated with the above scores. EHNA was not efficacious, even at toxic levels, while AICA riboside potentiated the efficacy of MTX. The data suggests that efficacious MTX therapy in rat AA (1) blocks purine biosynthesis; (2) increases in in vivo AICA levels. Also adenosine accumulation and blockage of adenosine deaminase (i.e., by EHNA) appear to be less critical to MTX efficacy. Increased levels of AICA metabolites may suppress the immune response in rat AA. PMID- 17408935 TI - Angiotensin converting enzyme inhibitor captopril modifies conditioned place preference induced by morphine and morphine withdrawal signs in rats. AB - Angiotensin II and angiotensin converting enzyme (ACE) inhibitors have analgesic, anticonvulsant and antidepressive effects and in some cases they can antagonize morphine. In the present study effects of angiotensin II and ACE inhibitor captopril administered intracerobroventricularily (icv) on conditioned place preference induced by morphine as well as on morphine withdrawal signs has been evaluated in rats. Icv canullas were implanted in anesthetized male rats. Rats were allowed to recover from the surgery and conditioned place preference was induced by morphine, and the time spent in morphine compartment was compared in saline, morphine, captopril and Ang II groups. Morphine withdrawal signs were compared in three other groups of rats: morphine alone, captopril+morphine and Ang II+morphine 4 days after morphine injections (three times in each day) with naloxone injection on 4th day. Results with rats conditioned place preference induced by morphine showed that icv captopril decreased significantly the time in morphine compartment (P<0.01) while Ang II had no effect. In morphine dependent rats captopril decreased some withdrawal signs after naloxone precipitation (P<0.05 and P<0.01). Ang II administration augmented some of withdrawal signs than in the morphine group (P<0.01 and P<0.001). In conclusion captopril reduced conditioned place preference induced by morphine and some withdrawal signs in morphine dependent rats. PMID- 17408936 TI - Cross-linking of chitosan and chitosan/poly(ethylene oxide) beads: a theoretical treatment. AB - The major aim of this study was to get deeper insight into the process of polymer cross-linking and the resulting structure of beads based on chitosan (CS) or chitosan/poly(ethylene oxide) (CS/PEO) semi-interpenetrating networks (semi-IPNs) as new carrier materials for oral drug delivery. Spherical hydrogels were prepared by a dropping method. The uptake kinetics of the cross-linking agent glyoxal into the beads were monitored and quantitatively described using Fick's second law of diffusion. High-resolution synchrotron infrared microspectroscopy (SIRM) was used to characterize the inner structures of the beads. Importantly, the diffusion of glyoxal through the hydrogels was found to be much slower than the cross-linking reaction and the mesh size of the created networks to be much larger than the hydrodynamic diameter of glyoxal. The presence of PEO chains slightly decreased the diffusivity of glyoxal due to obstruction effects. However, the cross-linking reaction was not affected. Interestingly, the polymers were homogeneously cross-linked throughout the beads, except for a thin outer shell showing an elevated cross-linking density. Thus, the obtained cross-linked hydrogel-based beads exhibit well-defined polymeric structures and offer an interesting potential as novel oral drug delivery systems. PMID- 17408937 TI - A model describing the effect of enzymatic degradation on drug release from collagen minirods. AB - A drug delivery system, named minirod, containing insoluble non-cross-linked collagen was prepared to investigate the release of model drug compounds. To characterise the complete drug release process properly, a mathematical model was developed. Previously, a mathematical model describing water penetration, matrix swelling and drug release by diffusion from dense collagen matrices has been introduced and tested. However, enzymatic matrix degradation influences the drug release as well. Based on experimental data, a model was developed which describes drug release by collagenolytic matrix degradation based on enzyme diffusion, adsorption and cleavage. Data for swelling, collagen degradation and FITC dextran release from insoluble equine collagen type I minirods were collected. Sorption studies demonstrated a tight sorption of collagenase on collagen surfaces that follows a Freundlich sorption isotherm and results in a degradation constant of 3.8x10(-5) mol/l for the minirods. The diffusion coefficients of FITC dextran 20 and 70 (3x10(-3) and 2.4x10(-3) cm2/h) in water were analyzed by fluorescence correlation spectroscopy (FCS). Using these data, the mathematical model was verified by two-dimensional simulations. The numerical results agreed well with the measurements. PMID- 17408938 TI - Cell-based fluorescence assay for evaluation of new-drugs potential for phospholipidosis in an early stage of drug development. AB - To evaluate new-drugs potential for phospholipidosis (PL), we developed a cell based fluorescence assay using a fluorescent-labeled phospholipid analogue (NBD PE). CHL/IU cells derived from newborn hamster lung were exposed to positive reference compounds (amiodarone, imipramine, chloroquine, propranolol, chlorpromazine and amantadine) in the presence of NBD-PE, and the level of PL, as indicated by accumulation of fluorescent inclusions in the cytoplasm, was evaluated using fluorescence microscopy and fluorometry. All positive reference compounds induced accumulation of fluorescent inclusions in a concentration dependent manner with an increase in fluorescence intensity. Fluorescence microscopically, the positive dose of test compound was determined as the concentration with a grade equivalent to or above that of 3.13 microM of amiodarone. Based on this criterion, 8 of 20 test compounds including PL-positive or -negative compounds were judged positive that were concurrent with the pathological results from rat toxicity studies. Furthermore, a positive criterion for fluorometry was decided as equivalent to or above 25% of maximum intensity induced by 1.56-25.0 microM amiodarone. In comparison of fluorometry methods with fluorescence microscopy method, 19 of 20 compounds were judged same. From these findings, we concluded that the assay developed in this study is a rapid and reliable method to predict new-drugs potential for PL at an early stage of drug development. PMID- 17408939 TI - Analysis of the locomotor activity of a nocturnal desert lizard (Reptilia: Gekkonidae: Teratoscincus scincus) under varying moonlight. AB - 1. This project seeks to identify determinants of the variation observed in the foraging behavior of predatory animals, especially in moonlight, using a lizard as a model. 2. Moonlight generally enhances the foraging efficiency of nocturnal visual predators and often depresses the locomotor activity of prey animals. Previous evidence has indicated for three different nocturnal species of smallish gecko lizards that they respond to moonlight by increasing their activity. 3. In this study some aspects of the foraging activity of the somewhat larger nocturnal psammophilous Teratoscincus scincus, observed near Repetek and Ashgabat, Turkmenistan, were significantly depressed by moonlight, while several confounding factors (sex, maturity, size, sand temperature, hour, prior handling and observer effect) were taken into account. 4. This behavioral difference may relate to the eye size of the various species. 5. Additionally, a novel method of analyzing foraging behavior shows that in this species the duration of moves increases the duration of subsequent stationary pauses. Measurement of locomotor speed, yielding an average speed of 220% of the maximum aerobic speed, indicates a need for these pauses. Secondarily, pause duration decreases the duration of subsequent moves, precluding escalation of move duration. 6. The results of this and related projects advocate the taking into account of physiological and environmental factors that may affect an animal's foraging behavior. PMID- 17408940 TI - Characterization of enterococci populations in livestock manure using BIOLOG. AB - The BIOLOG system was used to generate knowledge of enterococci populations found in fresh and dry manure of livestock (cattle (Bos taurus), horse (Equus caballus), and sheep (Ovis aires)). Six-hundred and forty Enterococcus isolates from the host sources were observed as a combined fresh manure unit and a combined dry manure unit, E. casseliflavus and E. mundtii were predominant in fresh manure (36% and 35%, respectively) as well as in dry manure (51% and 28%, respectively). The other species were found at a frequency of less than 10%. A chi-square test of the two most predominant Enterococcus sp. indicated that there were some significant differences among the frequency of E. casseliflavus and E. mundtii in cattle and sheep, but not horse. Despite these differences, these two species were overwhelmingly predominant among all three livestock sources. PMID- 17408941 TI - Complications of lung resection and exercise capacity: a meta-analysis. AB - RATIONALE: While exercise capacity, expressed as maximal oxygen consumption (VO2max), has been proposed to be the best predictor of postoperative cardiopulmonary complications after surgical resection in lung cancer patients, the literature remains controversial. The purpose of this study was to use the meta-analytic approach to determine if VO2max, expressed as either ml kg(-1) min( 1) or as a percentage of predicted, differed between patients who develop postoperative cardiopulmonary complications versus those that do not. METHODS: Studies were retrieved via (1) computerized literature searches, (2) cross referencing from retrieved articles, and (3) expert review of our reference list. Trials were included if they reported preoperative VO2max values (ml kg(-1) min( 1) or percentage of predicted) and had patients in which postoperative cardiopulmonary complications occurred. RESULTS: Fourteen studies representing a total of 955 men and women met our criteria for inclusion. Across all designs and categories, random-effects modeling demonstrated that patients without postoperative pulmonary complications had significantly higher levels of VO2max in ml kg(-1) min(-1) (mean difference=3.0, 95% confidence interval (CI), 1.9-4.0) as well as VO2max as a percentage of predicted (mean difference=8, 95% CI, 3.3 12.8). CONCLUSION: After a systematic review of the literature, we found that exercise capacity, expressed as VO2max, is lower in patients that develop clinically relevant complications after curative lung resection. These results are important for the practicing clinician because they answer the literature controversy on the usefulness of measuring preoperative exercise capacity and reinforce the current guidelines on decision making for lung resection. PMID- 17408942 TI - A study on engineering characteristics of asphalt concrete using filler with recycled waste lime. AB - This study focuses on determining the engineering characteristics of asphalt concrete using mineral fillers with recycled waste lime, which is a by-product of the production of soda ash (Na(2)CO(3)). The materials tested in this study were made using a 25%, 50%, 75%, and 100% mixing ratio based on the conventional mineral filler ratio to analyze the possibility of using recycled waste lime. The asphalt concretes, made of recycled waste lime, hydrated lime, and conventional asphalt concrete, were evaluated through their fundamental engineering properties such as Marshall stability, indirect tensile strength, resilient modulus, permanent deformation characteristics, moisture susceptibility, and fatigue resistance. The results indicate that the application of recycled waste lime as mineral filler improves the permanent deformation characteristics, stiffness and fatigue endurance of asphalt concrete at the wide range of temperatures. It was also determined that the mixtures with recycled waste lime showed higher resistance against stripping than conventional asphalt concrete. It was concluded from various test results that a waste lime can be used as mineral filler and, especially, can greatly improve the resistance of asphalt concrete to permanent deformation at high temperatures. PMID- 17408943 TI - Temporary stabilization of air pollution control residues using carbonation. AB - Carbonation presents a good prospect for stabilizing alkaline waste materials. The risk of metal leaching from carbonated waste was investigated in the present study; in particular, the effect of the carbonation process and leachate pH on the leaching toxicity of the alkaline air pollution control (APC) residues from municipal solid waste incinerator was evaluated. The pH varying test was conducted to characterize the leaching characteristics of the raw and carbonated residue over a broad range of pH. Partial least square modeling and thermodynamic modeling using Visual MINTEQ were applied to highlight the significant process parameters that controlled metal leaching from the carbonated residue. By lowering the pH to 8-11, the carbonation process reduced markedly the leaching toxicity of the alkaline APC residue; however, the treated APC residue showed similar potential risk of heavy metal release as the raw ash when subjected to an acid shock. The carbonated waste could, thereby, not be disposed of safely. Nonetheless, carbonation could be applied as a temporary stabilization process for heavy metals in APC residues in order to reduce the leaching risk during its transportation and storage before final disposal. PMID- 17408944 TI - Allocation and 'what-if' scenarios in life cycle assessment of waste management systems. AB - Many modern waste treatment processes and waste management systems are able to treat many different types of waste at the same time, and deliver a number of useful outputs (secondary materials, energy) as well. These systems are thus increasingly multi-functional. As such, in life cycle assessment studies, they create problems related to multi-functionality and allocation. Especially in LCAs of waste management systems, the solution in the form of system expansion or avoided burdens approach dominates the practice, and the partitioning approach plays a minor role. In this paper, we analyse the logic and problems of these two approaches. It appears that for the avoided burdens approach, the number of 'what if' assumptions is so large that LCAs on the same topic lead to quite diverging results. Since 'what-if' questions cannot be answered in an unambiguous way, such questions should preferably be left outside of a primarily scientific tool. The partitioning approach is not free from arbitrary choices as well, but, in contrast to the 'what-if' approaches, it does not claim to predict what happens or what would have happened. PMID- 17408945 TI - A living cell-based biosensor utilizing G-protein coupled receptors: principles and detection methods. AB - This study explores the feasibility of using a bullfrog fibroblast cell line (FT cells) expressing G protein coupled receptors (GPCRs) as the basis for a living cell-based biosensor. We have fabricated gold microelectrode arrays on a silicon dioxide substrate that supports long term, robust growth of the cells at room temperature and under ambient atmospheric conditions. Activation of an endogenous GPCR to ATP was monitored with an optical method that detects rises in intracellular calcium and with an electrochemical method that monitors the increased secretion of pre-loaded norepinephrine on a MEMS device. FT cells were also transfected to express reporter genes driven by several different promoters, raising the possibility that they could be modified genetically to express novel GPCRs as well. The ability to harness GPCRs for BioMEMS applications by using cells that are easy to grow on MEMS devices and to modify genetically opens the way for a new generation of devices based on these naturally selective and highly sensitive chemoreceptors. PMID- 17408946 TI - A highly sensitive biosensor with (Con A/HRP)n multilayer films based on layer-by layer technique for the detection of reduced thiols. AB - The bilayer of Con A/HRP through the biospecific affinity of concanavalin A (Con A) and glycoprotein horseradish peroxidase (HRP) was prepared on the surface of an Au electrode modified by the precursor film consisted of poly(allylamine hydrochloride) poly(sodium-p-styrene-sulfonate). Atomic force microscopy and electrochemical impedance spectroscopy were adopted to monitor the uniform layer by-layer assembly of the Con A/HRP bilayers. The amperometric measurement was based on the inhibition of reduced thiols and performed in the presence of the electron mediator hydroquinone in 0.2M phosphate buffer of pH 6.5 at an applied potential of -0.15 V versus Ag/AgCl. Under the optimal conditions, the biosensor presented a linear response for cysteine from 0.1 to 23.5 microM, with a detection limit of 0.02 microM. The biosensor demonstrated high stability and repeatability. A series of reduced thiols were detected by this inhibition biosensor and oxidized thiols showed no effect on the current response of the biosensor. PMID- 17408947 TI - DNA hybridization detection with organic thin film transistors: toward fast and disposable DNA microarray chips. AB - We demonstrate a novel DNA hybridization detection method with organic thin film transistors. DNA molecules are immobilized directly on the surface of organic semiconductors, producing an unambiguous doping-induced threshold voltage shift upon hybridization. With these shifts, single-stranded DNA (ssDNA) and double stranded DNA (dsDNA) are differentiated successfully. This method is expected to result in higher sensitivity than the main competitive technology, ISFET-based sensors because of the direct exposure of DNA molecules to sensitive layers. Factors that influence sensor sensitivity have been analyzed and optimum conditions have been determined using statistically designed experiments. Under the optimum conditions, the maximum difference between saturation current ratios caused by ssDNA and dsDNA reaches as high as 70%. In order to make DNA detection fast, we also demonstrate rapid on-chip electrically enhanced hybridization using the TFTs. These technologies together will enable the realization of disposable, rapid-turnaround tools for field-deployable genomic diagnosis. PMID- 17408948 TI - Highly sensitive lactate biosensor by engineering chitosan/PVI-Os/CNT/LOD network nanocomposite. AB - A novel chitosan/PVI-Os(polyvinylimidazole-Os)/CNT(carbon nanotube)/LOD (lactate oxidase) network nanocomposite was constructed on gold electrode for detection of lactate. The composite was nanoengineered by selected matched material components and optimized composition ratio to produce a superior lactate sensor. Positively charged chitosan and PVI-Os were used as the matrix and the mediator to immobilize the negatively charged LOD and to enhance the electron transfer, respectively. CNTs were introduced as the essential component in the composite for the network nanostructure. FESEM (field emission scan electron microscopy) and electrochemical characterization demonstrated that CNT behaved as a cross linker to network PVI and chitosan due to its nanoscaled and negative charged nature. This significantly improved the conductivity, stability and electroactivity for detection of lactate. The standard deviation of the sensor without CNT in the composite was greatly reduced from 19.6 to 4.9% by addition of CNTs. With optimized conditions the sensitivity and detection limit of the lactate sensor was 19.7 microA mM(-1)cm(-2) and 5 microM, respectively. The sensitivity was remarkably improved in comparison to the newly reported values of 0.15-3.85 microA mM(-1)cm(-2). This novel nanoengineering approach for selecting matched components to form a network nanostructure could be extended to other enzyme biosensors, and to have broad potential applications in diagnostics, life science and food analysis. PMID- 17408949 TI - Always all information in the delivery room--paternalism the other way around? PMID- 17408950 TI - Spinal anesthesia for cesarean section in a woman with Kartagener's syndrome and a twin pregnancy. AB - Kartagener's syndrome is an inherited disease characterized by a triad of symptoms: bronchiectasis, situs inversus and sinusitis resulting from defective cilial motility. There are few reports in the literature regarding the optimum anesthetic technique in patients with Kartagener's syndrome. The main anesthetic considerations are related to the respiratory system and increased risk of infection. We report the case of a woman with Kartagener's syndrome and a twin pregnancy conceived by in-vitro fertilization-embryo transfer, who underwent cesarean section under spinal anesthesia. Despite recurrent pulmonary problems, the twin pregnancy resulted in a successful outcome. This was facilitated by a close working relationship between the obstetrician, anesthesiologist and patient. PMID- 17408951 TI - Purification and characterization of extracellular xylanase from Streptomyces cyaneus SN32. AB - Streptomyces cyaneus SN32 was used in this study to produce extracellular xylanase, an important industrial enzyme used in pulp and paper industry. The enzyme was purified to homogeneity by ammonium sulfate precipitation followed by anion exchange chromatography using DEAE-Sepharose column, with 43.0% yield. The enzyme was found to be a monomer of 20.5 kDa as determined by SDS gel electrophoresis and has a pI of 8.5. The optimum pH and temperature for purified xylanase activity was 6.0 and 60-65 degrees C, respectively. The half-lives of xylanase at 50 and 65 degrees C were approximately 200 and 50 min, respectively. The xylanase exhibited K(m) and V(max) values of 11.1 mg/ml and 45.45 micromol/min/mg. The 15 residue N-terminal sequence of the enzyme was found to be 87% identical up to that of endoxylanases from Steptomyces sp. Based on the zymogram analysis, sequence similarity and other characteristics, it is proposed that the purified enzyme from S. cyaneus SN32 is an endoxylanase and belongs to Group 1 xylanases (low molecular weight - basic proteins). The purified enzyme was stable for more than 20 week at 4 degrees C. Easy purification from the fermentation broth and its high stability will be highly useful for industrial application of this endoxylanase. PMID- 17408952 TI - Comparison of Penicillium echinulatum and Trichoderma reesei cellulases in relation to their activity against various cellulosic substrates. AB - Penicillium echinulatum has been identified as a potential cellulase producer for bioconversion processes but its cellulase system has never been investigated in detail. In this work, the volumetric activities of P. echinulatum cellulases were determined against filter paper (0.27 U/mL), carboxymethylcellulose (1.53 U/mL), hydroxyethylcellulose (4.68 U/mL), birchwood xylan (3.16 U/mL), oat spelt xylan (3.29 U/mL), Sigmacell type 50 (0.10 U/mL), cellobiose (0.19 U/mL), and p nitrophenyl-glucopiranoside (0.31 U/mL). These values were then expressed in relation to the amount of protein and compared those of Trichoderma reesei cellulases (Celluclast 1.5L FG, Novozymes). Both enzyme complexes were shown to have similar total cellulase and xylanase activities. Analysis of substrate hydrolysates demonstrated that P. echinulatum enzymes have higher beta glucosidase activity than Celluclast 1.5L FG, while the latter appears to have greater cellobiohydrolase activity. Unlike Celluclast 1.5L FG, P. echinulatum cellulases had enough beta-glucosidase activity to remove most of the cellobiose produced in hydrolysis experiments. However, Celluclast 1.5L FG became more powerful than P. echinulatum cellulases when supplemented with exogenous beta glucosidase activity (Novozym 188). Both cellulase complexes displayed the same influence over the degree of polymerization of cellulose, revealing that hydrolyzes were carried out under the typical endo-exo synergism of fungal enzymes. PMID- 17408953 TI - Beta-substituted cyclohexanecarboxamide cathepsin K inhibitors: modification of the 1,2-disubstituted aromatic core. AB - Further SAR study around the central 1,2-disubstituted phenyl of the previously disclosed Cat K inhibitor (-)-1 has demonstrated that the solvent exposed P2-P3 linker can be replaced by various 5- or 6-membered heteroaromatic rings. While some potency loss was observed in the 6-membered heteroaromatic series (IC(50)=1 nM for pyridine-linked 4 vs 0.5 nM for phenyl-linked (+/-)-1), several inhibitors showed a significantly decreased shift in the bone resorption functional assay (10-fold for pyridine 4 vs 53-fold for (-)-1). Though this shift was not reduced in the 5-membered heteroaromatic series, potency against Cat K was significantly improved for thiazole 9 (IC(50)=0.2 nM) as was the pharmacokinetic profile of N methyl pyrazole 10 over our lead compound (-)-1. PMID- 17408954 TI - Anticipatory postural adjustments modify the movement-related potentials of upper extremity voluntary movement. AB - To elucidate the effect on movement-related potentials (MRPs) of anticipatory postural adjustments (APAs) accompanied by voluntary focal movement, we examined the MRPs of shoulder flexion movement under standing and sitting postural conditions in 12 normal subjects. MRPs were evaluated based on three components: readiness potential (RP), motor potential (MP), and movement-monitoring potential. APAs were observed in the activities of postural muscles including the biceps femoris and erector spinae muscles only under standing conditions. The amplitudes of the three MRP components were larger under standing conditions than under sitting conditions for all recorded electrode positions, and the RP and MP amplitudes at the vertex position, which lies over the supplementary motor area (SMA), showed a prominent increase under standing conditions with the highest statistical significance. These results suggest that a recruited neural process of the cortical area including the SMA may be necessary to generate voluntary movement accompanied by APA. PMID- 17408955 TI - alpha-Synuclein and its disease-related mutants interact differentially with the microtubule protein tau and associate with the actin cytoskeleton. AB - alpha-Synuclein is a primarily neuronal protein that is enriched at the pre synapse. alpha-Synuclein and the microtubule binding protein tau have been implicated in neurodegenerative diseases. alpha-Synuclein is known to associate with phospholipid vesicles, regulates dopamine metabolism and exhibits chaperone activity, but its main role remains largely unknown. Furthermore, knowledge on its interactions and post-translational modifications is essential for a molecular understanding of alpha-synucleinopathies. We investigated alpha synuclein mutations, causative for autosomal dominant forms of Parkinson's disease (A30P, A53T and E46K), and phosphorylation mutants at serine 129 (S129A and S129D) using fluorescently labelled alpha-synuclein, actin and tau. The investigation of colocalization, and protein-protein interactions by Forster resonance energy transfer and fluorescence lifetime imaging showed that alpha synuclein associates with the actin cytoskeleton and interacts with tau. The A30P mutation and cytoskeletal destabilization decreased this interaction. Given the concurrent loss of membrane binding by this mutation, we propose a membrane-bound functional complex with tau that might involve the actin cytoskeleton. PMID- 17408956 TI - The flexible interrelation between AOX respiratory pathway and photosynthesis in rice leaves. AB - Alternative respiratory pathway was investigated in rice seedlings grown under total darkness, light/dark cycle, or continuous light. The capacity of the alternative pathway was relatively higher in leaves that had longer light exposure. An analysis of rice AOX1 multigene family revealed that AOX1c, but not AOX1a and AOX1b, had a light-independent expression. The alternative oxidase (AOX) inhibitor, salicylhydroxamic acid (SHAM, 1mM), inhibited nearly 68% of the capacity of the alternative pathway in leaves grown under different light conditions. The plants grown under different light periods were treated with SHAM and then were exposed to illumination for 4h. The transition from dark to 4h of light stimulated the capacity of alternative pathway in etiolated rice seedlings and in those grown under light/dark cycle, whereas the capacity of the alternative pathway was constant in seedlings grown under continuous light with additional 4h of illumination. Etiolated leaves did not show any CO(2) fixation after 4h of illumination, and the increase in chlorophyll content was delayed by the SHAM pretreatment. When seedlings grown under light/dark cycle were moved from dark and exposed to 4h of light, increases in chlorophyll content and CO(2) fixation rate were reduced by SHAM. Although these parameters were stable in plants grown under continuous light, SHAM decreased CO(2) fixation rate but not the chlorophyll content. These results indicate that the role and regulation of AOX in light are determined by the developmental stage of plant photosynthetic apparatus. PMID- 17408957 TI - Differential expression of Arabidopsis sulfurtransferases under various growth conditions. AB - Sulfurtransferases (Str) comprise a group of enzymes widely distributed in archaea, eubacteria, and eukaryota which catalyse the transfer of a sulfur atom from suitable sulfur donors to nucleophilic sulfur acceptors. Neither the in vivo sulfur donors nor the acceptors of Str could be clearly identified in any of the organisms investigated so far. In Arabidopsis thaliana 20 Str proteins have been identified and grouped according to sequence homology. To investigate their respective in vivo function, Arabidopsis plants were grown in sterile hydroponic cultures at different sulfate (50, 500, and 1500 microM) and phosphate (0.1 and 1mM) concentrations, and in medium supplemented with 1mM thiosulfate. Northern blot analysis revealed the differential expression of the Str investigated. Thiosulfate Str activity was significantly increased at low sulfate concentrations in the medium. The Str mRNA levels were highly dependent on the developmental stage of the Arabidopsis plants. The expression of most Str analysed increased with progressing plant age in parallel with increasing 3 mercaptopyruvate and thiosulfate Str activities. The Str investigated were differentially expressed in a light/dark cycle whereas Str enzyme activities were not affected by the light conditions. The results indicate that each Str is regulated in a different way and plays an individual specific role in the plant metabolism. PMID- 17408958 TI - Salinity effects on polyphenol content and antioxidant activities in leaves of the halophyte Cakile maritima. AB - Cakile maritima is a local oilseed halophyte exhibiting potential for secondary metabolite production. In the present study, plant growth, leaf polyphenol content and antioxidant activity were comparatively analyzed in two C. maritima Tunisian accessions (Jerba and Tabarka, respectively sampled from arid and humid bioclimatic stages) under salt constraint. Three-week-old plants were subjected to 0, 100, and 400 mM NaCl for 28 days under glasshouse conditions. A significant variability in salt response was found between both accessions: while Tabarka growth (shoot biomass, leaf expansion) was significantly restricted at 100 and 400 mM NaCl, compared to the control, Jerba growth increased at 100mM before declining at 400 mM NaCl. The better behaviour of Jerba salt-challenged plants, compared to those of Tabarka, may be related to their higher polyphenol content (1.56- and 1.3-fold the control, at 100 and 400 mM NaCl respectively) and antioxidant activity (smaller IC(50) values for both 1,1-diphenyl-2 picrylhydrazyl and superoxide scavenging), associated with lower leaf MDA accumulation (ca. -66% of the control at 100mM NaCl). Taken together, our findings suggest that halophytes may be interesting for production of antioxidant compounds, and that the accession-dependent capacity to induce antioxidative mechanisms in response to salt, may result in a corresponding variability for growth sustainability. PMID- 17408959 TI - Complete regression of a thymoma to glucocorticoids, commenced for palliation of symptoms. AB - A patient is presented with Type B1 thymoma that showed complete regression to oral glucocorticoids commenced for palliation of symptoms. The patient has remained symptom free without radiological recurrence after 12 months. PMID- 17408960 TI - Comparison of on pump and off pump coronary surgery: risk factors for neurological outcome. AB - OBJECTIVE: Cerebrovascular accidents (CVA) are devastating complications after coronary artery bypass grafting (CABG). The reported incidence of neurological complications after conventional CABG (CCABG) is 3-6%. Off-pump coronary bypass grafting (OPCAB) has been associated in recent studies to a decreased morbidity and risk of perioperative stroke. Nevertheless, uncertainty still surrounds the relative benefits of OPCAB. We investigated whether, in our experience, OPCAB was associated with lower neurological morbidity than conventional CABG approach. METHODS: Eight thousand and two patients underwent isolated CABG at our institution between January 1998 and January 2005. OPCAB operation was performed on 1415 patients. Data were prospectively collected. A multiple logistic regression analysis was used to evaluate the influence of the two different surgical techniques on the neurological outcomes. RESULTS: Patients in the OPCAB group were significantly older (66.2 vs 63.5%, p<0.0001), had a higher incidence of renal injury (5.4 vs 2.4%, p<0.0001), and were more redo interventions (6.95 vs 1.53%, p<0.0001). The CCABG patients were more urgent at operation (5.46 vs 3.26, p=0.0007), were less hypertensive (57.6 vs 63% of the patients, p=0.0003) more diabetics (22 vs 20.6%, NS), and had an ejection fraction less than 0.40 (10.4 vs 9.6%, NS). CVA incidence was similar in both groups (Type I outcome: OPCAB=0.70% vs CCABG=0.68%, p=0.91; Type II outcome OPCAB=0.70% vs CCABG=0.83%, p=0.63). CONCLUSIONS: In our experience patients undergoing CCABG were not exposed to a greater risk of neurological adverse events when compared to OPCAB patients. PMID- 17408962 TI - Allograft aortic root replacement in complex prosthetic endocarditis. AB - OBJECTIVE: To evaluate perioperative and long-term results of complex prosthetic valve endocarditis treated by allograft aortic root replacement. METHODS: From April 1988 through February 2006, 41 patients diagnosed as prosthetic valve endocarditis (PVE) complicated by root abscess and/or periprosthetic leak, underwent fresh allograft valve replacement by root replacement. There were 37 males (89.7%) and the mean age was 51.5+/-13.7 years. The NYHA functional class was 3.0+/-0.1. Thirty-seven patients (90%) had a mechanical prosthesis and in 10 (25.6%) the PVE was recent (< 3 months). Ten patients (24.4%) underwent emergency valve replacement and four (9.8%) presented with chronic renal failure. The patients were followed for a mean of 54.3 (2-166) months and the end-points were death of the patient or allograft failure. RESULTS: There were two hospital deaths (4.8%), both in patients with perioperative low cardiac output. Nine patients had transient acute renal failure (22.0%) but none required dialysis. Three patients (7.3%) needed pacemaker for complete A-V block. Eight patients (19.5%) died late; two died of cardiac reasons, four of non-cardiac reasons (stroke-one; acute colecystitis-two; traffic accident-two) and two of unknown cause. Two patients needed reoperation due to allograft failure at 61 and 82 months. In no case was there evidence of recurrence of endocarditis during the follow-up. The 10-year survival was 79%. CONCLUSIONS: Allograft aortic root replacement in prosthetic endocarditis complicated by abscess and/or periprosthetic leakage carries low morbidity and mortality and, in this series, no recurrence of infection. In our experience, these results are superior to those obtained with other valvular substitutes. PMID- 17408963 TI - Quality of life research in head and neck cancer: a review of the current state of the science. AB - Quality of life (QOL) has become an increasingly important outcome measure for patient's undergoing treatment for a wide array of illnesses. QOL is a global construct that reflects a patient's general sense of well being. It is by definition multi-dimensional and reflective of the patient's point of view. Health related issues are among the many factors that may influence QOL. Since head and neck cancer (HNC) affects structures that are critical for normal functions such as speech and swallowing, and treatment may lead to deformities that adversely impact psychosocial functioning, there is particular interest in assessing QOL in this cohort of patients. In order to interpret the HN QOL literature, it is important to have an understanding of the significance and limitations of QOL assessments in the head and neck patient population as well as an appreciation for the currently available measurement tools. Unfortunately, the HNC QOL literature has many limitations including: small sample size, lack of prospective data and poor study design. None-the-less, important insights can be obtained by review of the current literature. First, it is important that QOL studies be reported in such a way as to provide clinically meaningful data to clinicians. Linking measurements with clinical benchmarks is one way to accomplish this goal. In addition, both general and HNC specific measures are needed in order to provide a comprehensive assessment of global health and tumor specific issues. In general, QOL declines immediately after therapy and returns toward baseline by 1 year. Several factors have been identified that may predict for worse QOL outcomes including: the presence of a feeding tube, co-morbid disease, tracheotomy, site and stage. Data correlating QOL with functional outcome and symptom burden fails to demonstrate a consistent relationship. This may be attributed to methodological issues in study design or the patient's ability to adapt to functional and symptom control problems. Whether routine use of QOL measures in the clinical setting is beneficial to patients has yet to be determined. Further studies are warranted as currently available instruments may not be valid for repeated clinical use. PMID- 17408964 TI - Association between plasma angiotensin-converting enzyme level and radiation pneumonitis. AB - Angiotensin-converting enzyme (ACE) plays an important role in pulmonary fibrosis and may be involved in the development of radiation-induced lung damage. The objective of this study was to evaluate the predictive value of plasma ACE in radiation pneumonitis (RP). Patients with stage I-III lung cancer were treated with radiotherapy with or without chemotherapy. ACE levels were measured using enzyme-linked immunosorbent assay before radiotherapy (pre-RT) and when a median dose of 45 Gy (Range: 40-48 Gy) was reached (during-RT). The primary end point was > or = grade 2 RP. Statistic significances were evaluated with independent T test and chi-square. Thirty-nine patients were enrolled in this study, among which 33.3% experienced > or = grade 2 RP. ACE levels, either pre-RT or during RT, were significantly lower in the RP group than in the non-RP group (P=0.02 and 0.03, respectively). Nine out of the 19 patients (47.4%) with pre-RT ACE levels < or = 462 ng/mL experienced RP, versus 3 of 19 (15.8%) patients with ACE levels > 462 ng/mL (P=0.04). This study suggested that plasma ACE as a predictive factor for radiation pneumonitis deserves further study. PMID- 17408965 TI - Adjuvant effects of crystal proteins from a Mexican strain of Bacillus thuringiensis on the mouse humoral response. AB - In this study, we determined the adjuvant effects of the crystal (Cry) proteins, p130, p98, and p64-62, on the immune response of mice to both sheep red blood cells (SRBC) and ovalbumin (OVA). The administration of p130, p98, and p64-62 Cry proteins to Balb/c mice induced a significant (p<0.01) increase in the production of anti-SRBC antibody-secreting cells (ASC). The p64-62 Cry proteins demonstrated the best ability to induce the production of IgA and IgG antibodies to SRBC (p<0.05), and IgM, IgA, and IgG antibodies to OVA (p<0.05). Additionally, Cry proteins did not produce any side effects associated with their administration to Balb/c mice. We suggest the potential use of the p64-62 Cry proteins as adjuvants for the administration of heterologous antigens. PMID- 17408966 TI - The effects of arginine on protein binding and elution in hydrophobic interaction and ion-exchange chromatography. AB - Arginine is effective in suppressing aggregation of proteins and may be beneficial to be included during purification processes. We have shown that arginine reduces non-specific protein binding in gel permeation chromatography and facilitates elution of antibodies from Protein-A columns. Here we have examined the effects of arginine on binding and elution of the proteins during hydrophobic interaction (HIC) and ion- exchange chromatographies (IEC) using recombinant monoclonal antibodies (mAbs) and human interleukin-6. In the case of HIC, the proteins were bound to a phenyl-Sepharose column in the presence of ammonium sulfate (AS) with or without arginine and eluted with a descending concentration of AS. While use of 1 M AS in the loading buffer resulted in complete binding of the mAb, inclusion of 1 M arginine in loading and equilibration buffer, only when using low-substituted phenyl-Sepharose, resulted in weaker binding of the proteins. While decreasing AS concentration to 0.75 M resulted in partial elution of the mAB, elution was facilitated with inclusion of 0.5-1 M arginine. In the case of IEC, arginine was included in the loading samples. Inclusion of arginine during binding to the IEC columns resulted in a greater recovery and less aggregation even when elution was done in the absence of arginine. These results indicate that arginine enhances elution of proteins bound to the resin, suggesting its effectiveness as a solvent for elution in HIC and IEC. PMID- 17408967 TI - Functional expression of the keratinolytic serine protease gene sfp2 from Streptomyces fradiae var. k11 in Pichia pastoris. AB - We report the initial characterization and expression of sfp2, a gene encoding a keratinolytic serine protease from Streptomyces fradiae var. k11. Recombinant SFP2 was expressed in and secreted from the yeast Pichia pastoris with a final yield of 78 mg/L (136.2 U/mL caseinolytic activity) after 25 h of induction. The recombinant enzyme was purified using by ammonium sulfate precipitation and gel filtration chromatography to electrophoretic homogeneity, which was appropriately glycosylated and had a molecular mass of 26.0 kDa. The purified recombinant SFP2 was characterized. The optimal pHs and temperatures of SFP2 for proteolysis of casein and keratin azure were pH 10.0, 60 degrees C, and pH 9.0, 55 degrees C, respectively. SFP2 activity was stable from pH 3.0 to pH 11.0. The enzyme activity was inhibited by Co(2+) and Cr(3+) and enhanced by Ni(2+) and Cu(2+). The K(m) of 0.45 mmol/L and V(max) of 19.84 mmol/min mg were calculated using N succinyl-Ala-Ala-Pro-Phe-pNA as a substrate. We tested the activity of SFP2 with soluble and insoluble substrates; SFP2 was more specific for keratinous substrates compared with proteinase K and other commercial proteases. PMID- 17408968 TI - High-level expression and purification of recombinant human catalase in Pichia pastoris. AB - Catalase is one of the antioxidant enzymes and is involved in many pathophysiologic processes and human diseases. This study focused on high-level expression and purification of recombinant catalase in Pichia pastoris. The cDNA encoding catalase was cloned by RT-PCR from Fetal liver of Homo sapiens. After PCR and construction of expression vector pPIC9K-CAT, human catalase was expressed highly in P. pastoris yeast SMD1168 and secreted into the culture medium. The secreted catalase was purified to a purity of 95% by ammonium sulfate fractionation, anionic exchange-chromatography, and Macro-prep Ceramic Hydroxyapatite with a overall yield of 60%. This study provides a new method for large-scale expression and purification of recombinant protein catalase. PMID- 17408969 TI - A size filtration approach to purify low affinity complexes for crystallization. AB - Low affinity protein complexes are difficult to isolate and handle in crystallization experiments. Size-exclusion chromatography often does not allow purification of the homogeneous complex. Here we used a size-filtration approach for the purification and concentration of the 19 microM affinity complex of yeast Rab-GTPase and its guanine nucleotide disassociation inhibitor (GDI). The homogeneous protein complex solution was crystallized and the structure was solved using the molecular replacement method. The resulting model of the low affinity unprenylated Rab-GDI complex should reflect a transient Rab-GDI complex when GDI is bound to the membrane-anchored Rab protein and is poised to extract Rab to cytosol. PMID- 17408970 TI - Molecular cloning and characterization of a putative lipopolysaccharide-induced TNF-alpha factor (LITAF) gene homologue from Zhikong scallop Chlamys farreri. AB - LPS-induced TNF-alpha factor (LITAF) is a novel transcriptional factor that was first discovered in LPS-stimulated human macrophage cell line THP-1. LITAF can bind to TNF-alpha promoter to regulate its expression. The first scallop LITAF (named as CfLITAF) was cloned from Zhikong scallop Chlamys farreri by Expressed Sequence Tag (EST) and Polymerase Chain Reaction (PCR) techniques. The cDNA of CfLITAF was of 1240 bp and consisted of a 5' untranslated region (UTR) of 112 bp, a 3' UTR of 678 bp and an open reading frame (ORF) of 450 bp encoding a polypeptide of 149 amino acids with an estimated molecular mass of 16.08 kDa and theoretical isoelectric point of 6.77. A typical conserved LITAF-domain was identified in CfLITAF by SMART analysis. Homology analysis of the deduced amino acid sequence of CfLITAF with other known sequences by using the BLAST program revealed that CfLITAF was homologous to the LITAF from human and rat (Identity = 46%), cattle, horse, mouse and chicken (Identity = 48%), western clawed frog (Identity=42%), and zebrafish (Identity = 50%). The mRNA expression of CfLITAF in different tissues including haemocytes, muscle, mantle, heart, gill and gonad, and the temporal expression in haemocytes challenged by LPS or peptidoglycan (PGN) were measured by Real-time RT-PCR. CfLITAF mRNA transcripts could be detected in all tissues examined and be up-regulated in haemocytes after LPS challenge. No significant changes were observed after PGN stimulation. All these data indicated the existence of LITAF in scallop and also provided clue on the presence of TNF-alpha-like molecules in invertebrates. PMID- 17408971 TI - Sexual dimorphism revealed in the structure of the mouse brain using three dimensional magnetic resonance imaging. AB - A large variety of sexual dimorphisms have been described in the brains of many vertebrate species, including humans. Naturally occurring sexual dimorphism has been implicated in the risk, progression and recovery from numerous neurological disorders, including head injury, multiple sclerosis and stroke. Genetically altered mice are a key tool in the study of structure-function relationships in the mammalian central nervous system and serve as models for human neuropsychiatric and neurological disorders. However, there are a limited number of quantitative three-dimensional analyses of the adult mouse brain structures. In order to address limitations in our knowledge of anatomical differences, a comprehensive study was undertaken using full 3D magnetic resonance imaging (MRI) to examine sexual dimorphisms in the C57BL/6J whole mouse brain. An expected difference in overall brain size between the sexes was found, where male brains were 2.5% larger in volume than female brains. Beyond the overall brain size differences in the sexes, the following significantly different regions were found: males were larger in the thalamus, primary motor cortex and posterior hippocampus, while females were larger in posterior hypothalamic area, entorhinal cortex and anterior hippocampus. Using high-definition 3D MRI on a normal inbred mouse strain, we have mapped in detail many sex-associated statistically significant differences in brain structures. PMID- 17408972 TI - Symmetrical event-related EEG/fMRI information fusion in a variational Bayesian framework. AB - In this work, we propose a symmetrical multimodal EEG/fMRI information fusion approach dedicated to the identification of event-related bioelectric and hemodynamic responses. Unlike existing, asymmetrical EEG/fMRI data fusion algorithms, we build a joint EEG/fMRI generative model that explicitly accounts for local coupling/uncoupling of bioelectric and hemodynamic activities, which are supposed to share a common substrate. Under a dedicated assumption of spatio temporal separability, the spatial profile of the common EEG/fMRI sources is introduced as an unknown hierarchical prior on both markers of cerebral activity. Thereby, a devoted Variational Bayesian (VB) learning scheme is derived to infer common EEG/fMRI sources from a joint EEG/fMRI dataset. This yields an estimate of the common spatial profile, which is built as a trade-off between information extracted from EEG and fMRI datasets. Furthermore, the spatial structure of the EEG/fMRI coupling/uncoupling is learned exclusively from the data. The proposed data generative model and devoted VBEM learning scheme thus provide an un supervised well-balanced approach for the fusion of EEG/fMRI information. We first demonstrate our approach on synthetic data. Results show that, in contrast to classical EEG/fMRI fusion approach, the method proved efficient and robust regardless of the EEG/fMRI discordance level. We apply the method on EEG/fMRI recordings from a patient with epilepsy, in order to identify brain areas involved during the generation of epileptic spikes. The results are validated using intracranial EEG measurements. PMID- 17408973 TI - Functional neuroimaging correlates of mental fatigue induced by cognition among chronic fatigue syndrome patients and controls. AB - The neural mechanisms underlying feelings of fatigue are poorly understood. The primary purpose of the study was to use functional magnetic resonance imaging (fMRI) to determine the association between feelings of mental fatigue and blood oxygen level dependent (BOLD) brain responses during a mentally fatiguing cognitive task. Healthy, non-fatigued controls and chronic fatigue syndrome (CFS) patients were included to determine the influence of chronic levels of fatigue on brain responses. We hypothesized that mental fatigue would be significantly related to brain activity during a fatiguing cognitive task but not during either a non-fatiguing motor (finger tapping) or cognitive (auditory monitoring) task. Patients (n=9) and controls (n=11) completed a finger tapping task, a simple auditory monitoring task and a challenging working memory task, designed to induce mental fatigue, while undergoing fMRI. Fatigue was measured prior to scanning and following each task during fMRI data collection. Results showed that mental fatigue was significantly related to brain activity during the fatiguing cognitive task but not the finger tapping or simple auditory monitoring tasks. Significant (p< or =0.005) positive relationships were found for cerebellar, temporal, cingulate and frontal regions. A significant (p=0.001) negative relationship was found for the left posterior parietal cortex. CFS participants did not differ from controls for either finger tapping or auditory monitoring tasks, but exhibited significantly greater activity in several cortical and subcortical regions during the fatiguing cognitive task. Our results suggest an association between subjective feelings of mental fatigue and brain responses during fatiguing cognition. PMID- 17408974 TI - Effects of motor fatigue on human brain activity, an fMRI study. AB - The main purpose of this study was to investigate effects of motor fatigue on brain activation in humans, using fMRI. First, we assessed brain activation that correlated with muscle activity during brief contractions at different force levels (force modulation). Second, a similar analysis was done for sustained contractions inducing motor fatigue. Third, we studied changes in brain activation due to motor fatigue over time. And fourth, we investigated cross-over effects of fatigue by comparing brain activation before and after the fatiguing condition during simple and high-order motor tasks (reaction time tasks). Several motor areas in the brain showed increased activity with increased muscle activity, both during force modulation and motor fatigue. Interestingly, the cerebellum showed a smaller increase in activation, during compensatory activation due to fatigue, while additional activation was found in the pre supplementary motor area and in a frontal area. During motor fatigue, there was a decrease in force production, an increase in force variability, and an increase in muscle activity. Brain areas comparable with the aforementioned areas also showed stronger activation over time. After fatigue, reaction time task performance remained the same (compared to before fatigue), while increased activation in orbitofrontal areas was found. Furthermore, there was a reduction in subjects' maximal voluntary contraction force, accompanied by a decrease in activation of the supplementary motor area (SMA). These results suggest that especially the activity in the SMA and frontal areas is affected by motor fatigue. PMID- 17408975 TI - Global phylogeography of the band-rumped storm-petrel (Oceanodroma castro; Procellariiformes: Hydrobatidae). AB - Factors shaping population differentiation in low latitude seabirds are not well understood. In this study, we examined global patterns of DNA sequence variation in the mitochondrial control region of the band-rumped storm-petrel (Oceanodroma castro), a highly pelagic seabird distributed across the sub-tropical and tropical Atlantic and Pacific Oceans. Despite previous classification as a single, monotypic species, fixed haplotype differences occurred between Atlantic and Pacific populations, and among all Pacific populations. In addition, Cape Verde and Galapagos birds formed distinct clades, estimated to have diverged from all other populations at least 150,000years ago. Azores hot season populations were also genetically distinct, lending support to previous phenotypic evidence that they be recognized as a separate species. Seasonal populations in Madeira probably represent separate genetic management units. The phylogeography of the band-rumped storm-petrel appears to have been shaped by both nonphysical barriers to gene flow and Pleistocene oceanographic conditions. Ancestral populations likely expanded through contiguous range expansion and infrequent long-distance colonization into their current breeding range. These findings suggest several possible revisions to the taxonomy of the band-rumped storm-petrel. PMID- 17408976 TI - Anatomy of the pectoralis minor tendon and its use in acromioclavicular joint reconstruction. AB - Many procedures described for operative management of acromioclavicular joint separations entail transfer of the coracoacromial ligament. We sought to describe the anatomy and morphology of the pectoralis minor tendon better, to assess its anatomic potential as a substitute for sacrificing the coracoacromial ligament, and to compare the ultimate tensile strength of the pectoralis minor with that of the coracoacromial ligament and detached coracoclavicular ligament. The morphology of the pectoralis minor tendon was carefully delineated and compared with that of the coracoacromial ligament, and 10 paired fresh-frozen cadaveric shoulders were tested to failure by applying a single uniaxial tensile load. Anatomic study of the pectoralis minor tendon confirmed its adequacy as a source of local autograft tissue in acromioclavicular joint reconstruction. We hypothesize that, in cases of acromioclavicular joint separation necessitating operative intervention, the use of the pectoralis minor tendon as a potential source of autograft tissue is anatomically feasible and it is slightly stronger than the coracoacromial ligament. PMID- 17408977 TI - The influence of medial reattachment of the torn cuff tendon for retracted rotator cuff tears. AB - We reattached the torn rotator cuff medial to the anatomic cuff insertion site if it was retracted. The purpose was to correlate the amount of medial reattachment of the cuff with shoulder function. We evaluated 63 shoulders in which repaired cuffs were followed with magnetic resonance imaging at a mean follow-up of 8 years. The amount of medial reattachment of the cuff tendon was determined by use of a T2-weighted oblique coronal view, which passed through the center of the humeral head, and was defined as the NCA angle (where N indicates the new cuff insertion point, C indicates the center of the humeral head, and A indicates the anatomic cuff insertion point). Theoretically, the more medially the cuff tendon was reattached, the greater the NCA angle. Neither the Japanese Orthopaedic Association shoulder score nor isometric strength of forward elevation was correlated with the NCA angle. The NCA angle was significantly correlated (P = .001) with the active forward elevation angle, which dramatically decreased at 30 degrees of the NCA angle, approximately 13 mm from the original cuff insertion point, assuming a humeral head radius of 25 mm. PMID- 17408978 TI - Acromioclavicular reconstruction using autogenous semitendinosus tendon graft: results of revision surgery in chronic cases. AB - Revision surgery of acromioclavicular dislocation is challenging owing to the altered anatomic relationships and the lack of stabilizing structures. In this study, an autogenous semitendinosus tendon graft was used for revision acromioclavicular stabilization, aiming at anatomic coracoclavicular reconstruction, as these patients had previously undergone a Weaver-Dunn procedure, which failed. Twelve patients were followed up clinically and radiographically for a mean of 49.5 months. The primary diagnosis was acromioclavicular joint dislocation Rockwood type III in 6, type IV in 4, and type V in 2 cases. At follow-up, the mean Constant score averaged 76.4 points. Pain relief was statistically significant (P < .01). Radiologic coracoclavicular distance and posterior displacement of the lateral clavicle in the Rockwood type IV cases decreased significantly (P < .01). We conclude that with this new technique of autogenous semitendinosus tendon graft replicating the anatomic ligamentous properties, good to excellent results can be achieved in revision cases of acromioclavicular reconstruction. PMID- 17408979 TI - Morphologic analysis of the distal humerus with special interest in elbow implant sizing and alignment. AB - This study determined the relationship between the medullary canal axis and the flexion-extension axis of the distal humerus as they relate to implant selection and design for elbow arthroplasty. Computed tomography scans of 40 fresh-frozen cadaveric specimens were analyzed with computer-aided design software. The anterior offset and cubital angle were measured between the 2 axes, and the cross sectional area and diameter were measured for the medullary canal at various intervals. The anterior offset of the flexion-extension axis from the medullary canal axis was proportional to the length of canal used to determine the stem axis. No correlation was established among the width of the articular surface, anterior-posterior canal curvature, and cubital angle. These findings suggest that modular implants that allow for the variability in the natural anterior bow and articular offset of the distal humerus may enhance proper restoration of the flexion-extension axis of the elbow. PMID- 17408980 TI - Bone mineral density in institutionalised patients with refractory epilepsy. AB - Bone mineral density (BMD) and content (BMC) were measured in 208 institutionalised patients with refractory epilepsy. BMD was in the osteoporotic range in 43% of males and 21% of females. BMC was reduced only in male patients. Apart from age and gender, no other factor or specific AED was particularly associated with reduced BMD or BMC. This study indicates the magnitude of the problem of bone disease in refractory epilepsy, in particular affecting young and male patients with chronic epilepsy. PMID- 17408981 TI - Long term response to steroid therapy in Rasmussen encephalitis. AB - Rasmussen encephalitis (RE) is a severe and progressive focal epilepsy of unknown origin that leads to deterioration of motor and cognitive function. In a previous study, we described positive effect of high doses of steroids during the first year after the onset of RE. The objective of this study was to evaluate this therapy at long term. We reviewed 11 patients (7 girls and 4 boys) with RE of the right hemisphere (7) and the left (4) at a follow-up of 9+/-2 years. Age at onset of RE ranged from 2 to 14 years. Six patients had no benefit from steroid therapy and underwent hemispherotomy. Five had significant reduction of seizure frequency with disappearance of epilepsia partialis continua, and improved motor function. Of these, two died of unexpected sudden death 5 and 7 years after seizure control. Two others with initial response experienced progressive recurrence of seizures 1 to 4 years after the end of steroid therapy and required hemispherotomy. Finally, only one patient exhibited total cessation of seizures with steroids for 3 years, but seizures progressively recurred although the frequency was moderate. Our data confirm that although steroid treatment can be useful when given early in the course of RE, long term relapse can occur among the good responders requiring delayed hemispheric disconnection. PMID- 17408982 TI - Pyridoxine induces non-specific EEG alterations in infants with therapy resistant seizures. AB - PURPOSE: In infants with frequent therapy resistant seizures (TRS-infants), clinical detection of pyridoxine-dependency (PD) or -responsiveness (PR) occurs by empirical intravenous (IV) pyridoxine administration during recording of the EEG. However, in undiagnosed TRS-infants it is still unclear to what extent EEG alterations by pyridoxine-IV are attributable to PD/PR or to non-specific responses. Before EEG alterations by pyridoxine-IV can be ascribed to PD/PR, these non-specific responses should be excluded first. METHODS: In 10 TRS-infants under 1 year of age, we determined the EEG effect by pyridoxine-IV on the EEG recording. RESULTS: After pyridoxine-IV administration, our data indicate declined (10-15%; p<0.05) EEG-amplitudes and total power (magnitude/frequency band) at frontal, central and centro-temporal electrodes. CONCLUSION: In TRS infants, pyridoxine-IV affects EEG-amplitude and -total power in a non-specific way, which does not identify PD/PR. PMID- 17408983 TI - Alleviation of osteoarthritis by calycosin-7-O-beta-D-glucopyranoside (CG) isolated from Astragali radix (AR) in rabbit osteoarthritis (OA) model. AB - OBJECTIVE: This work was undertaken to assess the protective effect of an isoflavonoid, calycosin-7-O-beta-D-glucopyranoside (CG), isolated from Astragali radix (AR) on the pathogenesis of osteoarthritis (OA)-like lesion in a rabbit model. METHODS: Nine rabbits underwent an anterior cruciate ligament and menisectomy transection (ACLMT) of the rear knee joints to induce OA-like lesion. They were randomly divided into three groups (n=6/group): a negative control group treated with 200 microl of 0.5% (v/v) dimethyl sulfoxide (DMSO), a positive control group treated with 200 microl of 100 microM piroxicam, and a test group treated with 100 microg/500 microl of CG, where the test agents were administered by injection once a week for 4 weeks starting from the third week. Rabbits were then sacrificed to observe the progression of OA-like lesion. The synovial fluid was analyzed for the amounts of total proteins, glycosaminoglycans (GAG) and prostaglandin E(2) (PGE(2)). In addition, histopathologic analyses were performed on the OA-like articular cartilage with or without therapeutic treatments. RESULTS: The total synovial fluid volume (P<0.05) was most strikingly reduced by the treatment with CG. Moreover, the CG treatment also significantly alleviated the OA-induced accumulation of prostaglandin (PG) (P<0.001) and total proteins (P<0.001) in the synovial fluid. The histopathologic analyses revealed that the CG treatment reduced the severity of the OA-like structural damages in the cartilage. However, the level of PGE(2), a pathologic inflammatory molecule, was not diminished by CG or piroxicam. CONCLUSION: These results indicate that the isoflavonoid CG isolated from AR significantly alleviated the pathologic changes in the OA-like rabbit knee joints. This suggests that CG from AR could be a promising treatment for the therapy of OA. PMID- 17408984 TI - Cross-cultural adaptation and validation of Singapore English and Chinese Versions of the Oxford Knee Score (OKS) in knee osteoarthritis patients undergoing total knee replacement. AB - OBJECTIVE: To cross-culturally adapt and validate Singapore English and Chinese versions of the Oxford Knee Score (OKS) in patients with knee osteoarthritis (OA) undergoing total knee replacement (TKR) surgery. METHODS: Singapore English and Chinese OKS versions were cross-culturally adapted from the source English version following standard guidelines (including cognitive debriefing), and validated by interviewing patients in English or Chinese using an identical, pretested questionnaire containing the OKS, Short Form 36, and EQ-5D. Reliability was assessed using Cronbach's alpha, dimensionality using principal component factor analysis and item-total correlations, convergent and divergent construct validity by assessing six and three a priori hypotheses, respectively. RESULTS: The Singapore English and Chinese OKS were well accepted by patients in pilot testing. When administered to a consecutive sample of 127 English and 131 Chinese speaking Singaporeans with knee OA (mean age 66 years, 83% female, mean duration of OA 6 years, Chinese:Malay:Indian:Others=78:7.9:11:3.1% for English version), Cronbach's alpha exceeded 0.8 and factor analysis yielded three factors for both versions. Hypothesized item-total correlations (Spearman's rho > or = 0.4) were observed for all items except limping, kneeling, and night knee pain in both versions. Convergent construct validity was supported by the presence of hypothesized moderate/strong correlations (rho=0.37-0.73) for six and five a priori hypotheses in English and Chinese versions, respectively. Divergent construct validity was supported by the presence of weak correlations (rho=0.09 0.30) for all three a priori hypotheses in both versions. CONCLUSION: Singapore English and Chinese OKS demonstrated good patient acceptability and psychometric properties (including construct validity) among multiethnic Asian patients with knee OA undergoing TKR. PMID- 17408985 TI - Evidence for JNK-dependent up-regulation of proteoglycan synthesis and for activation of JNK1 following cyclical mechanical stimulation in a human chondrocyte culture model. AB - OBJECTIVE: To examine the expression of mitogen-activated protein kinases (MAPKs) in human chondrocytes, to investigate whether selective activation of MAPKs is involved in up-regulation of proteoglycan (PG) synthesis following cyclical mechanical stimulation (MS), and to examine whether MS is associated with integrin-dependent or independent activation of MAPKs. METHODS: The C-28/I2 and C 20/A4 human chondrocyte cell lines were mechanically stimulated in monolayer cell culture. PG synthesis was assessed by [(35)S]-sulphate incorporation in the presence and absence of the p38 inhibitor SB203580, and the extracellular regulated kinase (ERK1/2) inhibitor PD98059. Kinase expression and activation were assessed by Western blotting using phosphorylation status-dependent and independent antibodies, and by kinase assays. The Jun N-terminal kinase (JNK) inhibitor SP600125 and the anti-beta(1) integrin (CD29) function-blocking antibody were used to assess JNK activation and integrin dependence, respectively. RESULTS: Increased PG synthesis following 3 h of cyclic MS was abolished by pretreatment with 10 microM SB203580, but was not affected by 50 microM PD98059. The kinases p38, ERK1/ERK2 and JNKs were expressed in both stimulated and unstimulated cells. Phosphorylated p38 was detected at various time points following 0.5, 1, 2 and 3 h MS in C-28/I2, but not detected in C 20/A4 cell lines. Phosphorylation of ERK1 and ERK2 was not significantly affected by MS. Phosphorylation of the 54 and 46 kDa JNKs increased following 0.5, 1, 2 and 3 h of MS, and following CO(2) deprivation. MS-induced JNK phosphorylation was inhibited by SB203580 at concentrations > or =5 microM and activation of JNK1 following MS was blocked by SP600125 and partially inhibited by anti-CD29. CONCLUSIONS: The data suggest JNK, rather than p38 or ERK dependent increases in PG synthesis, and selective, partially integrin-dependent, activation of JNK kinases in human chondrocyte cell lines following cyclical MS. JNK activation is also very sensitive to changes in CO(2)/pH in this chondrocyte culture model. PMID- 17408986 TI - Cerebrosides of baifuzi, a novel potential blocker of calcium-activated chloride channels in rat pulmonary artery smooth muscle cells. AB - Calcium-activated chloride channels (CaCCs) are crucial regulators of vascular tone by promoting a depolarizing influence on the resting membrane potential of vascular smooth muscle cells. However, the lack of a special blocker of CaCCs has limited the investigation of its functions for long time. Here, we report that CB is a novel potential blocker of I(Cl(Ca)) in rat pulmonary artery smooth muscle cells (PASMC). Cerebrosides (CB) were isolated from Baifuzi which is dried root tuber of the herb Typhonium giganteum Engl used for treatment of stroke in traditional medicine. Using the voltage-clamp technique, sustained Ca(2+) activated Cl(-) current (I(Cl(Ca))) was evoked by a K(+)-free pipette solution containing 500nM Ca(2+) which exhibited typical outwardly rectifying and voltage /time-dependence characterization. Data showed that CB played a distinct inhibitory role in modulating the CaCCs. Moreover, we investigated the kinetic effect of CB on I(Cl(Ca)) and found that it could slow the activation dynamics of the outward current, accelerate the decay of the inward tail current and change the time-dependence characterization. We conclude that CB is a novel potent blocker of CaCCs. The interaction between CB and CaCCs is discussed. PMID- 17408987 TI - Increased hormone levels in Tetrahymena after long-lasting starvation. AB - Tetrahymena contains vertebrate hormone-like materials. The level of one of these, insulin increased during starvation in a previous experiment. We hypothesized that other hormones are also influenced by starvation. To prove the hypothesis Tetrahymena pyriformis cultures were (1) starved for 24h; (2) starved for 24h and re-fed for 30min or (3) starved for 30min. Amount and localization of vertebrate-like hormones, produced by Tetrahymena, beta-endorphin, adrenocorticotropin (ACTH), serotonin, histamine, insulin and triiodothyronine (T(3)) were studied by immunocytochemical methods using flow cytometry and confocal microscopy. Long starvation elevated with 50% the hormone levels, while short starvation moderately elevated only the serotonin level in the cells. After short re-feeding endorphin and histamine returned to the basal level, ACTH and serotonin approached the basal level, however, remained significantly higher, while insulin and T(3) stood at the starvation level. The results show that such a stress as long starvation provokes the enhanced production of hormones which likely needed for tolerating the life-threatening effect of stress. PMID- 17408988 TI - Post-training reversible inactivation of the rat's basolateral amygdala interferes with hippocampus-dependent place avoidance memory in a time-dependent manner. AB - In this study, tetrodotoxin (TTX) inactivation was employed to evaluate the involvement of the rat's basolateral amygdala (BLA) in hippocampus-dependent spatial memory using a place avoidance learning task. Rats were trained in single 30 min session to avoid a 60 degrees segment of the stable circular (80-cm diameter) arena, entering which was punished by a mild shock. Bilateral injections of TTX or saline were made either immediately, 1 or 2h after training. Retention was tested 24h later in a 30 min extinction session. Retention was impaired when both BLA inactivated immediately or 1h after training, but not 2h after training. These data indicate that activity in the BLA, at least 60 min after training, is necessary for the post-training processing of a hippocampus dependent place avoidance memory. PMID- 17408989 TI - The feasibility of a percutaneous temporary pedal bypass. AB - A technique is described in which by subintimal recanalization it is possible to create a percutaneous distal bypass to the foot. This technique might offer new opportunities for patients with non-healing ulcers which are technically and/or medically inoperable. PMID- 17408990 TI - Residual venous reflux after superficial venous surgery does not predict ulcer recurrence. AB - OBJECTIVE: The aim of this study was to investigate the importance of venous reflux in ulcer recurrence following saphenous surgery. METHODS: Ulcerated legs (CEAP 5 and 6) with saphenous reflux were treated with superficial venous surgery plus compression as part of a clinical trial. Patients unfit for general anaesthesia (GA) underwent limited surgery under local anaesthesia (LA). Reflux in superficial and deep segments and venous refill times (VRTs) were assessed before surgery and 3-12 months post-operatively using duplex and digital photoplethysmography respectively. RESULTS: Of 185 patients treated with surgery, 15 failed to heal and 26 did not have a follow-up duplex. Within 3 years, 25 of the remaining 144 patients (17%) developed ulcer recurrence. Using a Cox regression model, the presence of residual venous reflux and change in reflux pattern were not found to be risk factors for ulcer recurrence (p=ns). LA was used in 4/25 patients who recurred compared to 28/119 who did not (p=0.60; Chi square test). For legs with recurrence, median VRT before surgery was 10.5s (range 5-29) compared to 11s (range 6-36) after surgery (p=0.097, Wilcoxon Signed Rank test). However, in legs without recurrence, median VRT increased from 10s (range 3-48) to 15s (range 4-48) after surgery (p<0.001). CONCLUSION: Residual reflux following saphenous surgery is not the most important predictor of venous ulcer recurrence. Poor venous function as demonstrated by VRT may be a better predictor of recurrence in these patients. PMID- 17408991 TI - Measurements of cerebrospinal fluid concentrations of S100beta protein during and after thoracic endovascular stent grafting. AB - OBJECTIVES: Thoracic endovascular aortic repair is associated with postoperative spinal cord ischemia in approximately 1 to 12.5% of all cases. S100beta is a protein that is released during acute damage of the central nervous system. This study was performed to determine the concentration of S100beta in cerebrospinal fluid during and after stenting of the thoracic aorta in patients at high risk for spinal cord ischemia. DESIGN: Prospective clinical study. MATERIALS AND METHODS: Eight patients who underwent elective thoracic aortic stent grafting underwent lumbar spinal fluid drainage. These patients were at high risk to develop spinal cord ischemia. METHODS: CSF samples for analysis of S100beta protein were drawn after induction of anesthesia, during stenting, once every hour the following six hours and 20 hours after repair. RESULTS: No significant increase in S100beta protein could be detected in CSF and no neurological deficits were detected postoperatively. CONCLUSIONS: The results of this study show us that there is no significant release of S100beta protein in CSF during stenting of the thoracic aorta in this subgroup of patients at high risk for spinal cord ischemia, consistent with clinical exam that there was no significant damage to the central nervous system. PMID- 17408992 TI - Is infrainguinal bypass grafting successful following failed angioplasty? AB - OBJECTIVES: Angioplasty is often used in the management of lower limb ischaemia and can reduce the need for infrainguinal bypass in some patients. There is an associated failure rate with this technique and bypass surgery is often used in this situation as a secondary limb salvage procedure. We aimed to evaluate the outcome of infrainguinal bypass grafting following failed attempt at angioplasty. METHODS: All cases of infrainguinal bypass at a single centre over a seven year period were identified and notes reviewed. Cases were divided into four groups according to their indication for surgery; acute ischaemia, chronic critical ischaemia, failed angioplasty and an 'other' group including aneurysmal disease and claudicants. The failed angioplasty group was compared with the other three groups. Survival analysis was performed using Kaplan Meier curves and groups compared in terms of long term patency and survival. RESULTS: Primary patency was 61.2% in the failed angioplasty group at 12 months compared with 60.6% in the other groups (P=1.11). There was also no significant difference in primary patency at 60 months (50% vs 40.6%, P=0.26). Survival at 12 months was also comparable between the groups (failed angioplasty group 74.2% compared with 77.3% in the other groups, P=0.662) as was 60 months survival (33.3% and 35.4% respectively, P=0.166). DISCUSSION: In this study, outcome of infrainguinal bypass following failed angioplasty was comparable to outcome of surgery performed for another indication. This paper supports the use of distal bypass surgery for limb salvage in cases where minimal access techniques have failed. PMID- 17408993 TI - Chronic venous disease in a cohort of healthy UK Asian men. AB - OBJECTIVES: This group has previously reported that UK Asians are significantly less likely to undergo surgery for lower limb venous disease than age and sex matched Caucasians. The aim of the present study was to estimate the prevalence of lower limb chronic venous disease (CVD) in the UK Asian male population. DESIGN: A prospective, epidemiological survey. MATERIALS AND METHODS: 100 unselected Asian men attending a local Mosque were assessed for the evidence of lower limb CVD, involving the collection of data on history and clinical signs and objective assessments of venous pathophysiology using lower limb venous ultrasonography and venous photoplethysmography (PPG). RESULTS: On clinical examination, 80 limbs (in 50 subjects) had clinical evidence of CVD, the majority of cases consisting of varicose veins (CEAP C2). No limbs had either healed or active ulceration (C5/6), and only 2 limbs had thread veins (C1). Eight subjects had had previous venous surgery. The venous refill time (vRT) measured by PPG was lower in limbs with CVD. On venous ultrasound, reflux was present in 73/200 limbs, affecting primarily the GSV system, with only 7 limbs having deep venous reflux. CONCLUSIONS: Present data strongly suggest that the low rates of superficial venous surgery in UK Asians is not because they are inherently less likely to develop CVD. PMID- 17408994 TI - Calf mortality as a welfare indicator on British cattle farms. AB - The objective of this study was to test calf mortality as an indicator of on-farm welfare and its use for welfare targeted surveillance. Calf mortality data were retrieved for three UK counties to estimate calf mortality rates at holding and county level. A selection criterion based on upper quartiles of calf mortality for the county of concern was defined. Its predictive ability was tested in a field study. The death risk of calves less than 6 months of age in 2002 was 1.76% in Inverness, 5.83% in Cheshire and 4.8% in Norfolk. Fifty-two welfare inspections matched by parish were conducted between October 2004 and January 2005. The positive predictive value was 26.92% and the negative predictive value was 65.38%. The addition of herd type, county and membership to an assurance scheme improved the predictive value. This study shows that calf mortality can be the starting point to design targeted welfare inspections in countries with centralized animal data recording systems. PMID- 17408995 TI - Bovine lymphotropic herpesvirus and non-responsive post-partum metritis in dairy herds in the UK. AB - Bovine lymphotropic herpesvirus (BLHV) was detected for the first time in the UK in December 2005 in a dairy herd suffering from chronic, non-responsive post partum metritis (NPPM). A small-scale investigation was undertaken in order to determine whether this was an isolated case. Samples of vaginal exudates or vaginal swabs were collected from cows in 13 UK dairy herds with a history of post-partum metritis that had not responded to standard treatment regimes for this condition. Cows in 9/13 herds and 1/13 herds were positive for BLHV and bovine herpesvirus-4, respectively, by pan-herpesvirus polymerase chain reaction. No consistent pattern of infectious agents or nutritional/metabolic factors commonly associated with post-partum metritis was observed at the times of sampling. The detection of BLHV in association with NPPM indicates that further work is warranted to determine the impact this virus has on cattle health. PMID- 17408996 TI - The effect of physical exercise on the daily rhythm of platelet aggregation and body temperature in horses. AB - The goal of this study was to investigate the influence of physical activity on the daily rhythm of platelet aggregation and body temperature in horses. Blood samples from 12 Thoroughbred horses, six sedentary animals and six athletes (studied both before and after a period of inactivity) were collected at 4h intervals for 48h via an intravenous cannula inserted into the jugular vein. Body temperature was recorded every 4h for 48h with a rectal probe. Platelet aggregation was measured with an aggregometer. Collagen was used to test the aggregation of the plasma samples. Statistical analysis of the data was performed by one-way repeated-measures analysis of variance (ANOVA) and by single cosinor method. Cosinor analysis identified the periodic parameters and their acrophases (expressed in hours) during the 2 days of monitoring. On each single day, there was a highly significant effect of time in all the horses, with P values <0.05. Temperature rhythms were unaffected by exercise. Platelet aggregation in exercising horses differed from the sedentary horses, and this difference disappeared after a 2-week period of rest. The results could be interpreted as indicating that physical exercise has an influence on the daily rhythm of platelet aggregation in horses. PMID- 17408997 TI - Protective effect of CpG-DNA against mastitis induced by Escherichia coli infection in a rat model. AB - A mastitis model in rats, induced by Escherichia coli infection, was established and the protective effect of Cytosine-phosphate-Guanosine (CpG)-DNA was determined. An E. coli suspension containing either 2 x 10(3) colony forming units (CFU)mL(-1)(EL group), 2 x 10(5)CFU mL(-1) (EH group), or (as controls) 100 microL phosphate buffer saline (CON group), was inoculated into the mammary glands 72 h after parturition. The rats were euthanased 24 h post-infection. The histopathological changes in mammary tissue in the EL group were mild, whereas the structural changes in the EH group were severe and polymorphonuclear leukocytes (PMNs) had accumulated in the mammary alveoli. Interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha and N-acetyl-beta-d-glucosaminidase (NAGase) were significantly increased in the mammary tissue from the EH group but not significantly changed in the EL group. On the basis of these findings, the potential protective effect of CpG-DNA on mammary glands was tested using a 2 x 10(5)CFU mL(-1) suspension. An intramuscular injection of either CpG-DNA (200 microg) or PBS (100 microL) was given immediately after parturition. At 72 h post partum, 2 x 10(5)CFU mL(-1)E. coli (100 microL) were inoculated into the mammary glands of all rats. At pre-infection (0 h), and 8, 16, 24, 48 and 72 h after inoculation six rats were euthanased. CpG-DNA induced more rapid migration of PMNs from the blood to mammary tissue at the initial stage of infection, stimulated the secretion of IL-6 and TNF-alpha at different time points, reduced viable E. coli in mammary tissues and decreased the activity of NAGase. CpG-DNA also promoted the expression of its specific receptor TLR-9 mRNA in mammary tissue. The study showed that CpG-DNA protected against E. coli mastitis in this rat model. PMID- 17408998 TI - Evolution of cortical metabolic abnormalities and their clinical correlates in Sturge-Weber syndrome. AB - BACKGROUND: The natural course of Sturge-Weber syndrome (SWS) is poorly understood, although neurological symptoms are often progressive. AIMS: To track longitudinal changes in brain glucose metabolism measured with positron emission tomography (PET) and their relation to clinical changes during the early course of SWS. METHODS: Fourteen children (age 3 months to 3.9 years at enrollment) with SWS and unilateral leptomeningeal angioma underwent two consecutive glucose metabolism PET scans with a mean follow-up time of 1.2 years. Longitudinal changes of the extent of cortical glucose hypometabolism on the angioma side were measured and correlated with age, clinical seizure frequency and hemiparesis. RESULTS: An increase in the size of the hypometabolic cortex was seen in 6 children, coinciding with an age-related increase in cortical glucose metabolism measured in unaffected contralateral cortex. These 6 patients were younger both at the initial (mean age 0.75 vs. 2.8 years; p<0.001) and the second scan (mean age 1.8 vs. 4.2 years; p=0.001) than those with no change in the extent of hypometabolic cortex (n=6). The area of cortical hypometabolism decreased in the two remaining children, and this was associated with resolution of an initial hemiparesis in one of them. Seizure frequency between the two scans was higher in children who showed progressive enlargement of cortical hypometabolism, as compared to those with no progression (p=0.008). CONCLUSIONS: In SWS, detrimental metabolic changes occur before 3 years of age coinciding with a sharp increase of developmentally regulated cerebral metabolic demand. Progressive hypometabolism is associated with high seizure frequency in these children. However, metabolic abnormalities may remain limited or even partially recover later in some children with well-controlled seizures. Metabolic recovery accompanied by neurological improvement suggests a window for therapeutic intervention in children with unilateral SWS. PMID- 17408999 TI - General parameter relations for the Shinnar-Le Roux pulse design algorithm. AB - The magnetization ripple amplitudes from a pulse designed by the Shinnar-Le Roux algorithm are a non-linear function of the Shinnar-Le Roux A and B polynomial ripples. In this paper, the method of Pauly et al. [J. Pauly, P. Le Roux, D. Nishimura, A. Macovski, Parameter relations for the Shinnar-Le Roux selective excitation pulse design algorithm, IEEE Transactions on Medical Imaging 10 (1991) 56-65.] has been extended to derive more general parameter relations. These relations can be used for cases outside the five classes considered by Pauly et al., in particular excitation pulses for flip angles that are not small or 90 degrees. Use of the new relations, together with an iterative procedure to obtain polynomials with the specified ripples from the Parks-McClellan algorithm, are shown to give simulated slice profiles that have the desired ripple amplitudes. PMID- 17409000 TI - Transcorneal tube extrusion in a child with a Baerveldt glaucoma drainage device. AB - We describe a clinical scenario in which a child was treated for glaucoma and aqueous misdirection with surgical placement of a tube shunt. The child then suffered extrusion of the tube through the cornea, a rare complication of such procedures. This complication has only been seen in children, and thus, young patients with tube shunts need careful and frequent monitoring, especially during the first year of the post tube shunt period to ensure that they have not developed this complication. PMID- 17409001 TI - Change in proptosis following extraocular muscle surgery: effects of muscle recession in thyroid-associated orbitopathy. AB - PURPOSE: To evaluate the effect of strabismus surgery on proptosis in patients with thyroid-associated orbitopathy. METHODS: The medical records of 22 consecutive patients with thyroid-associated orbitopathy undergoing strabismus surgery were reviewed. Data pertaining to the number of muscles operated on, amount of muscle recession, prior orbital decompression, and exophthalmometry were evaluated. RESULTS: Thirty-eight eyes in 22 patients with thyroid-associated orbitopathy were studied before and after strabismus surgery. The mean change in exophthalmometry following strabismus surgery in all eyes was +0.6 mm (p < 0.01). Eyes with prior decompression averaged a 0.9 mm increase following strabismus surgery (p < 0.01); those without decompression averaged a 0.2 mm decrease (p = 0.658). In eyes that underwent two rectus muscle recessions the increase in Hertel measurements averaged 1.2 mm; when only one muscle was recessed, the average increase was 0.2 mm. In the eyes with muscle recession < or =5 mm, the mean exophthalmometric increase was 0.7 mm. When a muscle recession of more than 5 mm was performed, the exophthalmometry showed a mean increase of 0.5 mm. CONCLUSIONS: Strabismus surgery on patients with thyroid-associated orbitopathy can worsen proptosis, especially in those with prior decompression. When planning for orbital decompression, the surgeon should consider this effect. Moreover, patients should be made aware of the possible changes to their appearance. PMID- 17409002 TI - Membrane potential and H2O2 production in duodenal mitochondria from broiler chickens (Gallus gallus domesticus) with low and high feed efficiency. AB - Increased hydrogen peroxide (H2O2) production was observed in duodenal mitochondria obtained from broiler chickens with low feed efficiency (FE). As a decrease in mitochondrial membrane potential (Deltapsi(m)) due to mild uncoupling of oxidative phosphorylation reduces reactive oxygen species production, this study was conducted to evaluate the effect of uncoupling on Deltapsi(m) and H2O2 production in duodenal mitochondria isolated from broilers with low (0.48+/-0.02) and high (0.68+/-0.01) FE. Membrane potential and H2O2 production were measured fluorometrically and in the presence of different levels of an uncoupler, carbonylcyanide-p-trifluoromethoxyphenylhydrazone (FCCP). The Deltapsi(m) was higher (PT). The four oldest patients had irreversible brain damage prior to diagnosis. The youngest child, diagnosed and started on betaine therapy as a newborn, is healthy at her present age of three years. We compared biochemical data among four groups: 16 control subjects, eight heterozygous parents, and five affected children (for the latter group, both before and during treatment with betaine anhydrous). Plasma amino acid concentrations were used to estimate changes in cerebral methionine uptake resulting from betaine therapy. In all affected children, treatment with betaine (534+/-222 mg/kg/day) increased plasma S-adenosylmethionine, improved markers of tissue methyltransferase activity, and resulted in a threefold increase of calculated brain methionine uptake. Betaine therapy did not normalize plasma total homocysteine, nor did it correct cerebral 5-methyltetrahydrofolate deficiency. We conclude that when the 5-methyltetrahydrofolate content of brain tissue is low, dietary betaine sufficient to increase brain methionine uptake may compensate for impaired cerebral methionine recycling. To effectively support the metabolic requirements of rapid brain growth, a large dose of betaine should be started early in life. PMID- 17409007 TI - Prevalence of feline immunodeficiency virus infection in domesticated and feral cats in eastern Australia. AB - Serum samples from 340 pet cats presented to three inner city clinics in Sydney Australia, 68 feral cats from two separate colonies in Sydney, and 329 cattery confined pedigree and domestic cats in eastern Australia, were collected over a 2 year period and tested for antibodies directed against feline immunodeficiency virus (FIV) using immunomigration (Agen FIV Rapid Immunomigration test) and enzyme-linked immunosorbent assay methods (Snap Combo feline leukaemia virus antigen/FIV antibody test kit, IDEXX Laboratories). Western blot analysis was performed on samples in which there was discrepancy between the results. Information regarding breed, age, gender, housing arrangement and health status were recorded for all pet and cattery-confined cats, while the estimated age and current physical condition were recorded for feral cats. The FIV prevalence in the two feral cat populations was 21% and 25%. The majority of FIV-positive cats were male (60-80%). The FIV prevalence in cattery-confined cats was nil. The prevalence of FIV in the pet cat sample population was 8% (27/340) with almost equal prevalence in 'healthy' (13/170) and 'systemically unwell' (14/170) cats. The age of FIV-positive pet cats ranged from 3 to 19 years; all FIV-positive cats were domestic shorthairs with outside access. The median age of FIV-positive pet cats (11 years) was significantly greater than the median age of FIV-negative pet cats (7.5 years: P<0.05). The prevalence of FIV infection in male pet cats (21/172; 12%) was three times that in female pet cats (6/168; 4%; P<0.05). With over 80% of this pet cat population given outside access and continued FIV infection present in the feral population, this study highlights the need to develop rapid, accurate and cost-effective diagnostic methods that are not subject to false positives created by concurrent vaccination against FIV. This is especially important in re-homing stray cats within animal shelters and monitoring the efficacy of the new vaccine, which has not been challenged against Australian strains. The absence of FIV within cattery-confined cats highlights the value in routine screening and indoor lifestyles. This study provides cogent baseline FIV prevalences in three cat subpopulations which can be used for appraising potential disease associations with FIV in Australia. PMID- 17409008 TI - Tumor necrosis factor is critical to control tuberculosis infection. AB - Tumor necrosis factor (TNF) is critical and non-redundant to control Mycobacterium tuberculosis infection and cannot be replaced by other proinflammatory cytokines. Overproduction of TNF may cause immunopathology, while TNF neutralization reactivates latent and chronic, controlled infection, which is relevant for the use of neutralizing TNF therapies in patients with rheumatoid arthritis. PMID- 17409009 TI - Field-based evidence for the linkage of pfcrt and pfdhfr drug-resistant malaria genotypes and clinical profiles of severe malaria in Niger. AB - Drug resistance has been shown to increase malaria mortality and morbidity in both community- and hospital-based studies. We investigated the association between two Plasmodium falciparum drug resistance-related molecular markers and clinical profiles of severe malaria in children hospitalised in Niger. PCR-RFLP analysis showed that the codon 108 mutation of the pfdhfr gene was positively linked to severe malarial anaemia. These findings are consistent with persistent parasite infection leading to unbalanced anaemia in young children. No significant relationship was found between the molecular markers and hypoglycaemia or hyperparasitaemia. Conversely, the pfcrt T76 mutation was found to be negatively associated with cerebral malaria and neurological symptoms, such as convulsions and coma. These results have implications for the strain-specific virulence hypothesis and for parasite fitness and evolution. Our findings are discussed in regard to the local malaria transmission level. PMID- 17409010 TI - TRAF activation of C/EBPbeta (NF-IL6) via p38 MAPK induces HIV-1 gene expression in monocytes/macrophages. AB - C/EBPbeta plays a pivotal role in activation of human immunodeficiency virus type 1 (HIV-1) in monocytes/macrophages. However, mechanisms for functional regulation of C/EBPbeta remain uncharacterized. Previous studies indicated that NF-kappaB activation by tumor necrosis factor (TNF) receptor family, which activates TNF receptor associated factor (TRAF), induces HIV-1 expression. We found that TRAF signals activate HIV-1 LTR with mutations of NF-kappaB sites in promonocytic cell line U937, suggesting existence of an alternative HIV-1 activating pathway. In this study, we have characterized the signal transduction pathway of TRAF other than that leading to NF-kappaB, using U937 cell line, and its subline, U1, which is chronically infected by HIV-1. We show that signals downstream of TRAF2 and TRAF5 activate p38 MAPK, which directly phosphorylates C/EBPbeta, and that activation of p38 MAPK potently activates C/EBPbeta-mediated induction of HIV-1 gene expression. We also show TRAF2 and TRAF5 are expressed in monocytes/macrophages of spleen samples from HIV-1 infected patients. Identification of TRAF-p38 MAPK-CEBPbeta pathway provides a new target for controlling reactivation of latent HIV-1 in monocytes/macrophages. PMID- 17409011 TI - Oligonucleotide array analysis of cyclic tension-responsive genes in human periodontal ligament fibroblasts. AB - Mechanical stress results in differential gene expression that is critical to convert the stimulus into biochemical signals. Under physiological stress such as occlusal force, human periodontal ligament fibroblasts (HPLF) are associated with homeostasis of periodontal tissues however the changes in response to mechanotransduction remain uncharacterized. We hypothesized that cyclic tension responsive (CT) genes may be used to identify a set of fundamental pathways of mechanotransduction. Our goal was to catalogue CT genes in cultured HPLF. HPLF were subjected to cyclic tension up to 16h, and total RNA was isolated from both tension-loaded and static HPLF. The oligonucleotide arrays analysis revealed significant changes of mRNA accumulation for 122 CT genes, and their kinetics were assigned by the K-means clustering methods. Ingenuity Pathway Analysis was completed for HPLF mechanotransduction using 50 CT genes. This analysis revealed that cyclic tension immediately down-regulated all nuclear transcription factors except v-fos FBJ murine osteosarcoma viral oncogene homolog (FOS) reacting as an early responsive gene. In turn, transcription factors such as tumor protein p53 binding protein 2 (TP53BP2), and extra-nuclear molecules such as adrenergic receptor beta2 (ADRB2) were up-regulated after 1-2h, which may result in fundamental HPLF functions to adapt to cyclic tension. Subsequent inhibition assays using Y27632, a pharmacologic inhibitor of Rho-associated kinase (ROCK), suggested that HPLF has both ROCK-dependent and ROCK-independent CT genes. Mechanical stress was found to effect the expression of numerous genes, in particular, expression of an early responsive gene; FOS initiates alteration of HPLF behaviors to control homeostasis of the periodontal ligament. PMID- 17409012 TI - Differential expression of matrix metalloproteinases and tissue-derived inhibitors of metalloproteinase in fetal and adult skins. AB - Matrix metalloproteinases and their tissue-derived inhibitors determine the architecture of the extracellular matrix. In early gestation, the amount and organization of extracellular matrix may be associated with scarless repair of fetal skin wounds. To elucidate the part of the mechanism(s) underlying the phenotypic transition from scarless to scar-forming healing observed during fetal gestation, the ontogeny of matrix metalloproteinase-2, -9, -14 and their tissue inhibitors was characterized in non-wounded fetal human skin with different gestational ages from 13 to 33 weeks and adult skin using reverse transcriptase polymerase chain reaction, immunohistochemical staining and western blot protocols. We showed that the levels of gene expressions for matrix metalloproteinase-2, -9, -14 and their endogenous inhibitors were significantly more in late gestational and adult skins than that in early gestational skin. Similar results were noted in terms of protein contents of these enzymes and inhibitors in fetal and adult skins. We concluded that the endogenous matrix metalloproteinase-2, -9, -14 and their endogenous inhibitors might be involved in skin development and in maintenance of cutaneous structure and function. Lower protein contents of tissue-derived inhibitor-1, -2 in early gestational skin might provide a predominantly antiscarring signal while higher protein expression of these two inhibitors might be associated with scar-forming healing in late gestational and adult skins. PMID- 17409013 TI - Drugs and drug resistance in African trypanosomiasis. AB - Despite the many decades of use of most of the current trypanocides, we know little of their mode of action. This may in part be because most of these will act on multiple targets once inside the cell, and they derive their selective action on the parasite from selective accumulation by the pathogen. Loss of this capacity for drug uptake by the trypanosome would thus be a major cause for drug resistance. We here discuss the use of current drugs against human and veterinary African trypanosomiasis, the prevalence, causes and mechanisms of drug resistance and new developments in trypanosomiasis therapy such as the introduction of nifurtimox and DB289. PMID- 17409014 TI - Evaluation of mutagenic activity in supply water at three sites in the state of Rio Grande do Sul, Brazil. AB - In this study, the Salmonella/microsome assay, using the micro-suspension method, was utilized to evaluate water for public supply at three sites in Rio Grande do Sul. The first site selected was in an area under industrial influence and the others were in non-industrial reference areas. Based on 40 L samples of raw water and/or after conventional treatment, compounds were extracted with XAD4 resins using natural and acidic pH, and the extracts were analyzed in the TA98 and TA100 strains with and without S9. Raw water extracts in the industrial region induced 27.4 revertants/L (rev/L) for TA100+S9 up to 226.3 rev/L for TA100-S9, both for acidic pH extracts. After conventional treatment the responses varied from 20.6 rev/L (TA98-S9) for natural pH extracts to 755.5 rev/L (TA98-S9) for acidic pH extracts. For acidic extracts obtained from reference site samples, the response, with metabolic activation only, ranged from negative to minimal. Direct mutagenic responses in acidic extracts may be elevated in treated extracts, compared with raw water extracts, influenced by the presence of by-products of the chlorination process. However, the mutagenicity observed in the treated water extracts in the industrial area increased and reflected mainly the combination of directly and indirectly acting compounds in the source waters, that are heavily influenced by anthropogenic factors. PMID- 17409015 TI - Accumulation of macrophages expressing MRP8 and MRP14 in skin lesions during Leishmania major infection in BALB/c and RAG-2 knockout mice. AB - Migration inhibitory factor-related protein 8 (MRP8) and MRP14 are expressed by myeloid cells and especially known as marker proteins of an immature and inflammatory subtype of macrophages. In this study, we immunohistochemically examined an accumulation of MRP8+ and MRP14+ macrophages in skin lesions during Leishmania major infection in susceptible BALB/c and RAG-2-/- mice. L. major infection caused the development of a nodular type of skin lesion at the infection site in mice and a massive accumulation of macrophages was observed in the lesions at four weeks after the infection. Immunohistochemical analyses showed MRP8+ and MRP14+ macrophages are predominant cell types in the skin lesions in both mouse strains. In contrast, F4/80+ cells, which correspond to mature macrophages, were rarely found in the skin lesions. These data suggest that the accumulation of inflammatory subtype of macrophages in BALB/c mice during L. major infection can be induced without acquired immune responses. PMID- 17409016 TI - Clinicians' perceptions about use of computerized protocols: a multicenter study. AB - PURPOSE: Implementation of evidence-based techniques, such as explicit computerized protocols, has achieved limited success among clinicians. In this study, we describe the development and validation of an instrument for assessing clinicians' perceptions about use of explicit computerized protocols. METHODS: Qualitative assessment of semi-structured interviews with clinicians gave rise to a cognitive model evaluating the factors that motivate clinicians to use explicit computerized protocols. Using these constructs we developed a 35-item instrument which was administered to 240 clinicians (132 nurses, 53 physicians and 55 respiratory therapists), in three health-care institutions. RESULTS: Factor analysis identified nine factors that accounted for 66% of the total variance cumulatively. Factors identified were: Beliefs regarding Self-Efficacy, Environmental Support, Role Relevance, Work Importance, Beliefs regarding Control, Attitude towards Information Quality, Social Pressure, Culture, and Behavioral Intention. The strongest predictor was Beliefs regarding Self Efficacy, which accounted for 26% of the total variance of intention to use explicit computerized protocols. Results supported the reliability and construct validity of the instrument. CONCLUSIONS: Clinicians' perceptions play a critical role in determining their intention to use explicit computerized protocols in routine clinical practice. Behavioral theories will help us understand factors predicting clinicians' intention to use explicit computerized protocols and recognize the implications of these factors in the design and implementation of these protocols. PMID- 17409017 TI - Hemagglutinin pseudotyped lentiviral particles: characterization of a new method for avian H5N1 influenza sero-diagnosis. AB - BACKGROUND: Highly pathogenic avian influenza (HPAI) H5N1 has spread globally in birds and infected over 270 humans with an apparently high mortality rate. Serologic studies to determine the extent of asymptomatic H5N1 infection in humans and other mammals and to investigate the immunogenicity of current H5N1 vaccine candidates have been hampered by the biosafety requirements needed for H5N1 micro-neutralization tests. OBJECTIVE: Development of a serodiagnostic tool for highly pathogenic influenza that reproduces H5N1 biology but can be used with less biohazard. STUDY DESIGN: We have generated and evaluated H5 hemagglutinin pseudotyped lentiviral particles encoding the luciferase reporter (H5pp). RESULTS: H5pp entry into target cells depends on alpha2-3 cell surface sialic acids and requires low pH for membrane fusion. H5pp infectivity is specifically neutralized by sera from patients and animals infected with H5N1 and correlates well with conventional microneutralization test. CONCLUSIONS: H5pp reproduce H5N1 influenza virus entry into target cells and potentially provides a high throughput and safe method for sero-epidemiology. PMID- 17409018 TI - Combination therapy with ribavirin and amantadine in renal transplant patients with chronic hepatitis C virus infection is not superior to ribavirin alone. AB - BACKGROUND: Standard treatment of chronic hepatitis C virus (HCV) infection based on interferon is not an option in renal transplant recipients due to the high risk of acute allograft rejection. OBJECTIVES: To assess efficacy and tolerability of combined treatment with ribavirin and amantadine regarding viral clearance, normalization of liver enzymes, and improvement of HCV-related hepatopathy and graft nephropathy in HCV-RNA-positive renal transplant patients. STUDY DESIGN: Prospective randomized controlled study comparing ribavirin, 1000 mg daily (n=7), versus ribavirin, 1000 mg, in combination with amantadine, 200 mg daily (n=8), for 12 months, versus no therapy (controls, n=26). Results were evaluated by intention-to-treat analysis. RESULTS: No relevant differences among treatment groups were found regarding liver enzymes, HCV viremia, liver histology and renal parameters. However, antiviral treatment was limited by anemia, resulting in premature withdrawal from therapy and requiring substitution with recombinant erythropoietin in most patients. The best predictor for tolerability of active treatment was a creatinine clearance rate>50 ml/min. CONCLUSIONS: Addition of amantadine to ribavirin seems not to be superior to ribavirin monotherapy in renal transplant patients with chronic replicating HCV infection. However, this may be explained in part by the poor tolerability of both ribavirin and amantadine in patients with impaired renal function, resulting in drop-outs and subtherapeutic drug dosage. PMID- 17409019 TI - Risperidone-induced reduction in JC viruria as a surrogate marker for efficacy against progressive multifocal leukoencephalopathy and hemorrhagic cystitis. PMID- 17409020 TI - Intrafamilial hepatitis E in France. PMID- 17409021 TI - Display, engineering, and applications of antigen-specific T cell receptors. AB - The use of T cell receptors (TCRs) as potential therapeutic agents provides an opportunity to target a greatly expanded array of antigens, compared to those now targeted with monoclonal antibodies. With the advent of new display technologies and TCR formats for in vitro engineering, it should be possible to generate high affinity TCRs against virtually any peptide antigen that is shown to bind to a major histocompatibility complex (MHC) molecule (e.g. peptides derived from viral antigens or from self proteins that are associated with the transformed phenotype). What remains, however, are challenges associated with effective targeting of very low numbers of cell surface antigens (pepMHC), fewer than the case for conventional monoclonal antibody-based therapies. This hurdle might be overcome with the attachment of more effective payloads for soluble TCR approaches, or by using TCR gene transfer into T cells that can then be adoptively transferred into patients. There is considerable work to be done on the physiological aspects of either approach, including pharmacokinetic studies in the case of soluble TCRs, and T cell trafficking, persistence, and autoreactivity studies in the case of adoptively transferred T cells. As with the field of monoclonal antibodies, it will take time to explore these issues, but the potential benefits of TCR-based therapies make these challenges worth the effort. PMID- 17409022 TI - Occurrence of Legionella in hot water systems of single-family residences in suburbs of two German cities with special reference to solar and district heating. AB - A total of 452 samples from hot water systems of randomly selected single family residences in the suburbs of two German cities were analysed for the occurrence of Legionella. Technical data were documented using a standardized questionnaire to evaluate possible factors promoting the growth of the bacterium in these small plumbing systems. All houses were supplied with treated groundwater from public water works. Drinking water quality was within the limits specified in the German regulations for drinking water and the water was not chlorinated. The results showed that plumbing systems in private houses that provided hot water from instantaneous water heaters were free of Legionella compared with a prevalence of 12% in houses with storage tanks and recirculating hot water where maximum counts of Legionella reached 100,000 CFU/100ml. The presence of L. pneumophila accounted for 93.9% of all Legionella positive specimens of which 71.8% belonged to serogroup 1. The volume of the storage tank, interrupting circulation for several hours daily and intermittently raising hot water temperatures to >60 degrees C had no influence on Legionella counts. Plumbing systems with copper pipes were more frequently contaminated than those made of synthetic materials or galvanized steel. An inhibitory effect due to copper was not present. Newly constructed systems (<2 years) were not colonized. The type of hot water preparation had a marked influence. More than 50% of all houses using district heating systems were colonized by Legionella. Their significantly lower hot water temperature is thought to be the key factor leading to intensified growth of Legionella. Although hot water systems using solar energy to supplement conventional hot water supplies operate at temperatures 3 degrees C lower than conventional systems, this technique does not seem to promote proliferation of the bacterium. Our data show convincingly that the temperature of the hot water is probably the most important or perhaps the only determinant factor for multiplication of Legionella. Water with a temperature below 46 degrees C was most frequently colonized and contained the highest concentrations of legionellae. It is evident that the same factors affecting colonization by Legionella in large buildings also exist in small residential water systems. If temperatures are low there is no difference between large and small systems and Legionella counts are high in both. Since private residences are an important source of community-acquired legionellosis, these findings emphasize the need for preventive control measures in small residential buildings. In some situations it may be necessary to install filtration devices at the point-of-use. PMID- 17409023 TI - Nursing interventions to improve the health of men with prostate cancer undergoing radiotherapy: a review. AB - The aim of this study was to investigate what nurses do to improve the health of men who are receiving radiotherapy treatment due to prostate cancer. The method was a literature review using a systematic approach. The Cochrane Library, Medline and CINAHL databases were used in a search that covered the period from January 1994 to April 2006. The screening of 200 abstracts resulted in 14 articles corresponding to the research question, which were assessed according to scientific quality. Two independent reviewers performed the screening and quality assessment process using specific protocols. Two themes emerged: nurse-led care related to radiotherapy treatment and patients' experiences of radiotherapy treatment. The results show that there is strong scientific support for nurse-led follow-up care aimed at assisting patients by means of providing information on how to manage side effects (evidence grade A). In addition, there is moderate scientific support for the need to ensure that this information is structured, objective and concrete and that it can be provided by means of audiotapes or over the phone (evidence grade B) as well as weak scientific support for reporting patients' experiences of radiotherapy treatment (evidence grade C). PMID- 17409024 TI - Supraventricular tachycardia and the struggle to be believed. AB - BACKGROUND: Little research exists examining patient experiences of life with supraventricular tachycardia (SVT). Realistic expectations of symptoms, patient management strategies, and treatment options are important components of patient education and treatment decisions. AIM: This descriptive qualitative study explored patients' experiences living with SVT. METHODS: Grounded theory methodology was used with semi-structured interviews of 25 SVT patients undergoing radiofrequency ablation treatment. RESULTS: The main core category that emerged from the data was the "struggle to be believed". The struggle or work of living with SVT entails: (1) living with the uncertainty of the occurrence and duration of an episode, (2) "covering up" to manage symptoms and appear normal, (3) searching for causative factors to prevent further episodes, and (4) experimenting with management techniques to shorten or end episodes. CONCLUSION: Clinical diagnosis and referral for ablation treatment were more difficult for women suggesting existence of a gender bias. As with many other chronic conditions, the illness trajectory work is formidable. Patients in this study with SVT, however, reported facing the additional difficulty of being believed. Disruptions in self-esteem and personal relationships are the effects of not being believed. Providers could avoid minimizing women's symptoms by listening seriously and offering realistic expectations; including insight into patient experiences and the trajectory of the dysrhythmia; and efficacy of drug and ablation treatments. PMID- 17409025 TI - The influence of antiepileptic drugs on cognition: a comparison of levetiracetam with topiramate. AB - Levetiracetam (LEV) and topiramate (TPM) are considered highly effective novel antiepileptic drugs (AEDs) in the treatment of focal epilepsies. To explore potential side effects, this study investigated their influence on cognitive functions comparatively by means of a standardized neuropsychological test battery assessing several cognitive domains. In this observational study, cognitive changes were explored in 30 consecutively recruited patients with focal epilepsy treated with LEV and in 21 patients treated with TPM, comparing functions assessed prior to gradual initiation and after reaching steady state of the individual target dosage. Before titration, patient groups did not differ significantly with respect to cognitive performance. Whereas the LEV group manifested no change in cognitive performance after AED titration, the TPM group worsened in the cognitive domains of cognitive speed and verbal fluency, as well as short-term memory. These findings suggest that TPM, unlike LEV, may impair frontal lobe functions. The lack of cognitive side effects related to LEV treatment may be relevant for treatment decisions. PMID- 17409026 TI - Testing homogeneity of the risk ratio in stratified noncompliance randomized trials. AB - The risk ratio (RR), defined as the ratio of probabilities of having an adverse event between an experimental treatment and a control treatment, is one of the most commonly used indices to measure the efficacy of an experimental treatment in a randomized clinical trial (RCT). When we want to obtain a summary estimator of the RR in stratified analysis, it is important that we can examine the assumption whether the underlying RR varies between strata. Although we can find a few publications on testing the homogeneity of RR, all these papers focus the ideal situation in which every patient complies with his/her assigned treatment. The research on testing the homogeneity of RR for a noncompliance RCT is limited. In this paper, we develop three simple test statistics for testing homogeneity of the RR under a noncompliance stratified RCT with large strata. To evaluate the performance of these statistics, we apply Monte Carlo simulation to calculate Type I error and power in a variety of situations. Based on the findings, we provide a general guideline of selecting a preferable test statistic under various situations. Finally, we include a large field trial studying the effect of a multifactor intervention program on the mortality of coronary heart disease (CHD) in middle-aged men to illustrate the practical use of these test statistics. PMID- 17409027 TI - Monitoring of titanium base alloys-biofluids interface. AB - Monitoring of the titanium, Ti-5Al-4V, Ti-6Al-4Fe implant materials--Ringer 1 and Ringer 2 solutions (of different pH values) interface for long term was studied in this work. In Ringer 1 solution (with high chloride ion content) all biomaterials present self-passivation. On Ti-6Al-4Fe alloy, the breakdown of the passive film was registered but at high pitting potential; pitting protection potential is very noble and can not be reached in human fluids. In Ringer 2 solution was obtained more electropositive corrosion potential values than in Ringer 1 solution; pitting corrosion of Ti-6Al-4Fe alloy is characterised by nobler breakdown and pitting protection potential values, therefore a better pitting corrosion resistance and tendency. Ion release increases in time, for the first 400-600 immersion hours and then tend to a constant level with very low values, non-dangerous for human body. All open circuit potentials oscillate around some electropositive values. The potential gradients calculated for extreme pH values have low values during 20,000 exposure hours and can not accelerate the corrosion. Atomic Force Microscopy images obtained after different exposure periods in Ringer 1 solution revealed that the roughness increased in time, suggesting a dynamic process at biomaterial-biofluid interface. X-ray Photoelectron spectra obtained after 2880 immersion hours in Ringer 2 solution show the existence of protective titanium dioxide TiO(2) and TiO and Ti(2)O(3) oxides both for titanium and Ti-5Al-4V alloy. Also, Al(2)O(3) oxide was detected. PMID- 17409028 TI - Exploring the roles of Mus81-Eme1/Mms4 at perturbed replication forks. AB - Cells of all living organisms have evolved complex mechanisms that serve to stabilise, repair and restart stalled, blocked and broken replication forks. The heterodimeric Mus81-Eme1/Mms4 structure-specific endonuclease appears to play an important role(s) in homologous recombination-mediated processing of such perturbed forks. This enzyme has been implicated in the cleavage of stalled and blocked replication forks to initiate recombination, as well as in the processing of recombination intermediates that result from repairing damaged forks. In this review we assess the biochemical and genetic evidence for the mitotic role of Mus81-Eme1/Mms4 at replication forks and in repairing post-replication DNA damage. Mus81 appears to act when replication is impeded by genotoxins or by impairment of the replication machinery, or when arrested replication forks are not adequately protected. We discuss how its action is regulated by the S-phase cell cycle checkpoint, depending on the nature of the stalled or damaged fork. We also present a new way in which Mus81 may limit crossing over during the repair of post-replication gaps, and explore Mus81's interplay with other components of the recombination machinery, including the RecQ helicases that also play important roles in processing replication and recombination intermediates. PMID- 17409029 TI - Increased glucose excursion in cystic fibrosis and its association with a worse clinical status. AB - BACKGROUND: Abnormal glucose tolerance is a frequent co-morbidity in cystic fibrosis patients (CF), and is associated with a worse prognosis. The objectives are to investigate (a) the relative contribution of insulinopenia and insulin resistance (IR) for glucose tolerance and (b) the association between various glucose parameters and CF clinical status. METHODS: Oral glucose tolerance tests were performed in 114 consecutive CF patients not known to be diabetic as well as 14 controls similar for age and BMI. RESULTS: Abnormal glucose tolerance was found in 40% of patients with CF: 28% had impaired glucose tolerance (IGT) and 12% had new cystic fibrosis related diabetes (CFRD). Compared to control subjects, all CF patients were characterized by an increased glucose excursion (AUC). While reduced early insulin release characterised CF, IGT and CFRD patients also present IR thus both mechanisms significantly contribute to glucose tolerance abnormalities. Increased glucose AUC and reduced early insulin release but not glucose tolerance categories were associated with a reduced pulmonary function (FEV(1)). CONCLUSION: In CF, early insulin secretion defect but also IR contribute to glucose intolerance. Early in the course of the disease, increased glucose AUC and reduced early insulin secretion are more closely associated with a worse clinical status than conventional glucose tolerance categories. PMID- 17409030 TI - AMP-activated protein kinase and chemotransduction in the carotid body. AB - AMP-activated protein kinase (AMPK) is a key component of a kinase cascade that regulates energy balance at the cellular level. Our recent research has raised the possibility that AMPK may also function to couple hypoxic inhibition of mitochondrial oxidative phosphorylation to O(2)-sensitive K(+) channel inhibition and hence underpin carotid body type I cell excitation. Thus, in addition to maintaining the cellular energy state AMPK may act as the primary metabolic sensor and effector of hypoxic chemotransduction in type I cells. These findings provide a unifying link between two previously separate theories pertaining to O(2)-sensing in the carotid body, namely the 'membrane hypothesis' and the 'mitochondrial hypothesis'. Furthermore, our data suggest that in addition to its effects at the cellular level the AMPK signalling cascade can mediate vital physiological mechanisms essential for meeting the metabolic needs of the whole organism. PMID- 17409031 TI - Development of a RP-HPLC method for screening potentially counterfeit anti diabetic drugs. AB - Pharmaceutical counterfeiting is becoming a serious problem in the world, especially in developing countries including China. Herein an isocratic reversed phase high performance liquid chromatography (RP-HPLC) method was developed for screening counterfeit medicines and adulterated dietary supplement products. The developed method could be employed to separate and determine simultaneously six anti-diabetic drugs (glipizide, gliclazide, glibenclamide, glimepiride, gliquidone, repaglinide) on an isocratic solvent system using an Alltima C18 column (5 microm, 150 mmx4.6 mm) with an isocratic mobile phase of methanol phosphate buffer (pH 3.0; 0.01 mol/L) (70:30, v/v), at a flow rate of 1.0 mL/min and at a wavelength of 230 nm. The proposed method was successfully applied to the analysis of medicinal and dietary supplement samples purchased from the local market in China. PMID- 17409032 TI - Development and validation of a liquid chromatography-mass spectrometry assay for the determination of pyronaridine in human urine. AB - A reliable method has been developed for the determination of pyronaridine in human urine using amodiaquine as an internal standard. Liquid-liquid extraction was used for sample preparation. Analysis was performed on a Shimadzu LCMS-2010 in single ion monitoring positive mode using atmospheric pressure chemical ionization (APCI) as an interface. The extracted ion for pyronaridine was m/z 518.20 and for amodiaquine was m/z 356.10. Chromatography was carried out using a Gemini 5 microm C18 3.0 mmx150 mm column using 2 mM perflurooctanoic acid and acetonitrile mixture as a mobile phase delivered at a flow rate of 0.5 mL/min. The mobile phase was delivered in gradient mode. The retention times of pyronaridine and amodiaquine were 9.1 and 8.1 min respectively, with a total run time of 14 min. The assay was linear over a range of 14.3-1425 ng/mL for pyronaridine (R2>or=0.992, weighted 1/Concentration). The analysis of quality control samples for pyronaridine at 28.5, 285, 684 and 1140 ng/mL demonstrated excellent precision with relative standard deviation of 5.1, 2.3, 3.9 and 9.2%, respectively (n=5). Recoveries at concentrations of 28.5, 285, 684 and 1140 ng/mL were all greater than 85%. This LC-MS method for the determination of pyronaridine in human urine has excellent specifications for sensitivity, reproducibility and accuracy and can reliably quantitate concentrations of pyronaridine in urine as low as 14.3 ng/mL. The method will be used to quantify pyronaridine in human urine for pharmacokinetic and drug safety studies. PMID- 17409033 TI - Determination of eprosartan in human plasma and urine by LC/MS/MS. AB - A protein precipitation, liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the determination of eprosartan in human plasma and urine. The solvent system also served as a protein precipitation reagent. The chromatographic separation was achieved on a CAPCELL PAK C18 column (50 mmx2.0 mm, 5 microm, Shiseido). A mobile phase was consisted of 0.5% formic acid in water and 0.5% formic acid in acetonitrile (72:28). Detection was by positive ion electrospray tandem mass spectrometry on a Sciex API3000. The standard curves, which ranged from 5 to 2000 ng/mL in human plasma and from 0.25 to 50 microg/mL in urine, were fitted to a 1/x weighted quadratic regression model. The method proved to be accurate, specific and sensitive enough to be successfully applied to a pharmacokinetic study. PMID- 17409034 TI - Analytical method for the quantitative determination of cyanuric acid as the degradation product of sodium dichloroisocyanurate in urine by liquid chromatography mass spectrometry. AB - A simple and selective analytical method for the quantitative determination of cyanuric acid, the degradation product of sodium dichloroisocyanurate (NaDCC), in human urine is reported herein. The sample preparation involved the use of diatomaceous earth extraction columns. Quantification was achieved by liquid chromatography mass spectrometry using negative ion electrospray with a cyano (CN) column. Between day relative standard deviation less than 10% (n=6) was obtained at the 5 mg L(-1) level. The assay was linear over the investigated range 0-20 mg L(-1) and the limit of detection (LOD) was confirmed to be 0.1 mg L(-1). The method was applied to monitoring levels of cyanuric acid in healthcare workers using disinfectants products containing NaDCC. PMID- 17409035 TI - An effective sample preparation approach for screening the anticancer compound piceatannol using HPLC coupled with UV and fluorescence detection. AB - Piceatannol, compared with the renowned resveratrol, is a better anticancer agent and a superior agent with other biological activities. However, as there are only few plants reported to contain minute quantity of piceatannol, the scarcity of sources greatly impedes the piceatannol-related researches. To explore new sources of piceatannol, we established a sample preparation approach for screening the piceatannol in plants using HPLC-UV-fluorescence detection. When the HPLC is coupled with UV and fluorescence detectors, the decrease of signals in interferences and increase of signal in piceatannol in the fluorescence chromatogram mark clearly the position of the piceatannol peak; ultimately, it allows identification without standards. In this study, we systematically evaluated the factors affecting the extraction efficiency of piceatannol in sample preparation. Of the sample preparation strategies studied, direct solvent extraction and liquid nitrogen treatment followed by solvent extraction gave satisfactory recoveries for both piceatannol and resveratrol. These approaches avoided time-consuming lyophilization procedure. In addition, all procedures must be done in the dark to avoid negative impact of irradiation from fluorescence light on the recoveries of piceatannol and resveratrol. With the present method, we re-examined the plants previously claimed to contain only resveratrol for their piceatannol contents. The species examined included Polygonum cuspidatum, Arachis hypogaea, Vitis thunbergii, and Ampelopsis brevipedunculaata. The results showed, for the first time, all these plants contain piceatannol. The finding implies that the resveratrol-containing plants may also contain piceatannol. The results demonstrate the feasibility of these sample preparation approaches and may further the understanding for the distribution of piceatannol in plants. PMID- 17409036 TI - Surgical removal of an intrauterine device perforating the sigmoid colon: a case report. AB - BACKGROUND: Intrauterine devices (IUDs) are among the most common contraceptive methods in the world, but have been associated with extrauterine dislocation and bowel perforation. We report a case of an IUD perforation of the sigmoid colon. CASE REPORT: A 35-year-old woman with complaints of abdominal pain, fever, and diarrhea underwent exploratory laparotomy. During the procedure, a left tubo ovarian abscess was found as well as an IUD extruding through the uterine cavity with the two wings of the device entirely in the lumen of the sigmoid colon. The patient had total abdominal hysterectomy with bilateral salpingo-oophorectomy with resection of the sigmoid colon. CONCLUSION: This is a report of a symptomatic perforation by an IUD into the sigmoid colon after a period of 10years in situ. PMID- 17409037 TI - Surgical treatment of renal cell carcinoma with right atrial thrombus: early experience and description of a simplified technique. AB - Operative management of patients presenting renal cell carcinoma's (RCC) with right atrial tumor thrombus extension is a technical challenge. It requires the use of cardiopulmonary bypass (CPB). The aim of this study was to report our early experience and to describe a simplified CPB technique. 5 consecutive patients underwent surgical resection by a joint cardiovascular and urological team. The ascending aorta was canulated. The venous drainage was achieved using a proximal canula inserted in the superior vena cava and a distal canula inserted in the IVC below the renal veins. Right atrium thrombus extension was extracted under normothermic CPB without cross clamping or cardioplegic arrest. A cavotomy was performed at the ostium of the renal vein and an endoluminal occlusion catheter was introduced. The thrombectomy and the radical nephrectomy were then performed. The occurrence of gaseous or tumor embolism, operative time, perioperative bleeding, and post-operative complications were assessed. Mean patients age was 62.9 years. Atrial and caval thrombectomy were achieved successfully in all patients. Mean operative time was 206 min. Mean CPB time was 62 min. Mean hospital stay was 18.8 days. One death occurred, due to respiratory failure. An asymptomatic early thrombosis of the IVC was diagnosed by CT scan in 1 patient. The four remaining patients were alive 6 months after the surgical procedure. Minimally invasive CPB technique can be used to treat intra atrial thrombus tumor extension arising from renal cell carcinoma. It can be performed safely with acceptable complications rate. PMID- 17409038 TI - Functional reconstruction of large anogenital defects with bilateral V-Y advancement posterior thigh fasciocutaneous flaps. AB - BACKGROUND: In patients with perianal extramammary Paget's disease, wide local excision is recommended. In the reconstruction of the resulting large defects not only the closure of the defect but also the preservation of anogenital function need to be considered. METHODS: We used bilateral V-Y advancement posterior thigh fasciocutaneous flaps for reconstructing three large anogenital defects, including the total anal canal, resulting from wide local excisions attributed to extramammary Paget's disease. A temporary anus was constructed at the same time to allow the wound of the reconstructed anus to heal. After the surgery, the course of bowel function was followed up through interviews and manometric examinations. RESULTS: All defects were successfully reconstructed with the flaps, and bowel function recovered satisfactorily. Manometric examinations were carried out in two cases; anal pressures were recorded regularly and used as parameters of the recovery of bowel function before closing the artificial anus. Eventually, the patients recovered the function of defecation. CONCLUSIONS: We recommend the bilateral V-Y advancement posterior thigh fasciocutaneous flaps together with manometric studies as one of the most effective techniques for managing large anogenital defects. The use of sensate flaps, the restoration of original structures and manometric studies are conducive to successful outcomes. PMID- 17409039 TI - 'What's in a name?'. PMID- 17409040 TI - Management of bilateral Achilles tendon rupture associated with ciprofloxacin: a review and case presentation. AB - Fluoroquinolone antibiotics are increasingly being recognized as a cause of Achilles tendinitis and rupture. We report the case of a 62-year old man who developed bilateral Achilles tendon rupture six days following commencement of ciprofloxacin. Tendon exploration and repair was accomplished with the use of a prosthetic substitute (Leeds-Keio ligament) but healing was complicated by left wound breakdown that was successfully repaired with a perforator-based fasciocutaneous flap. A review of the current literature on fluoroquinolone associated achilles ruptures and the various methods of tendon and soft tissue management of the primary or complicated injuries are discussed. PMID- 17409041 TI - Successful resurfacing of scars from previous deliberate self-harm using Integra dermal matrix substitute. AB - BACKGROUND: There are currently limited treatment options available to modify the appearance of scarring due to deliberate self-harm (DSH). The authors present two cases of Integra dermal matrix use for resurfacing scars due to past DSH. METHODS: By excising the scars and resurfacing with Integra, we present and describe a novel technique for treating the visible stigmata of previous DSH. RESULTS: No complications occurred, and patients noted improved appearance, skin quality and self-esteem following surgery. Both patients would recommend this treatment to others. CONCLUSIONS: Integra dermal matrix substitute may be used successfully to treat scarring from previous DSH. PMID- 17409042 TI - Effects of the interaction between lean tissue mass and estrogen receptor alpha gene polymorphism on bone mineral density in middle-aged and elderly Japanese. AB - Because both genetic and environmental factors influence bone mass, it is important to examine the effect of gene-environment interactions on bone mineral density (BMD) for the prevention of osteoporosis at an individual level. Estrogen receptor alpha (ER alpha) plays an important role in increasing BMD via mechanical strain and muscle mass is a reflection of the forces the muscle applies to the bone. The aim of this study is to investigate the effect of the interaction between lean tissue mass (LTM) and the ER alpha polymorphisms T-->C (PvuII) [dbSNP: rs2234693] and A-->G (XbaI) [dbSNP: rs9340799] on BMD in middle aged and elderly individuals. Subjects were 2209 community-dwelling Japanese men and women, ages 40 to 79 years. ER alpha polymorphisms in the first intron, T-->C and A-->G were identified and lumbar spine and femoral neck BMD and LTM were measured by dual-energy X-ray absorptiometry. Both T-->C and A-->G polymorphisms were divided into two genotype groups (TT vs. TC/CC; AA vs. AG/GG). In postmenopausal women, the effect of LTM on femoral neck BMD was significantly larger for those with the TC/CC genotype than for those with the TT genotype for the T-->C polymorphism, and larger for those with the AG/GG genotype than for those with the AA genotype for the A-->G polymorphism. This gene-LTM interaction was observed at the femoral neck, but not at the lumbar spine. For men and premenopausal women, no gene-LTM interaction was found. In conclusion, there was an interaction between LTM and the ER alpha T-->C and A-->G polymorphisms with respect to their effect on femoral neck BMD in postmenopausal women and those with the TC/CC and AG/GG genotypes had larger effects of LTM than those with TT and AA genotypes. PMID- 17409043 TI - Effects of statins on bone mineral density: a meta-analysis of clinical studies. AB - CONTEXT: Statins inhibit HMG-CoA reductase, preventing synthesis of mevalonate but also of isoprenoids, which affect osteoclast activity. Amino-bisphosphonates share this effect. In vitro and in vivo, statins show convincing anabolic and anti-resorptive bone effects. However, in a clinical meta-analysis (MA), they did not prevent hip fractures. OBJECTIVE AND DESIGN: Our meta-analysis studied the impact of statins on bone mineral density (BMD) at various sites and compared the effects of lipophilic and more hydrophilic statins. DATA SOURCES: Our PubMed and Embase queries using two keywords (statins, BMD) were updated to October 2006. DATA COLLECTION: Two readers independently collected BMDs from studies. DATA SYNTHESIS: Twenty-one studies, mostly observational (three randomized controlled trials and one pseudo-randomized study), were assessed. Two studies were excluded (no control groups). Three studies could not be analyzed. The sixteen studies analyzed mainly included postmenopausal osteopenic women (2971 patients under statins). Statins significantly increased BMD at total hip (TH) and femoral neck (FN). Effect sizes (ESs) were modest: 0.21 at TH (95% confidence interval [CI]: 0.16-0.25) and 0.20 at FN (CI: 0.08-0.28). Among women, statins acted similarly (ES: 0.20 for TH and 0.18 for FN; CI: 0.14-0.25 and 0.06-0.31 respectively); lipophilic statins (simvastatin, lovastatin) almost entirely caused this effect, at both TH (ES: 0.20; CI: 0.15-0.26) and FN (ES: 0.22; CI: 0.06-0.37). CONCLUSION: Our findings of modest but statistically significant beneficial effects of statins on hip BMD should promote large double-blind randomized controlled trials on their bone effects, in view of their major beneficial cardiovascular effects with excellent safety profile. PMID- 17409044 TI - Resident satisfaction with long-term care services. PMID- 17409045 TI - A review of satisfaction instruments used in long-term care settings. AB - We review studies in the published literature using satisfaction surveys in long term care settings. These settings include nursing homes, assisted living facilities, hospital long-term care units, and residential care homes. Rather than simply describe instruments in existence, we compare and contrast the format, content, and administration of these instruments. We identified 50 studies that have either developed or used satisfaction instruments in long-term care facilities. However, we also show that satisfaction instruments in the published literature vary greatly in numerous ways, including the number of questions they contain, the number of respondents included in the study, assessment of psychometric properties, and in the number of domains used. PMID- 17409046 TI - The role of consumer satisfaction in ensuring quality long-term care: opportunities and challenges. AB - Consumer-satisfaction information can play a valuable role as one component among a broader set of publicly- and privately led activities to improve the quality of long-term care (LTC). However, measuring and using consumer-satisfaction information in LTC is not without its challenges. We start by defining the ways in which we use the term "consumer satisfaction" in this article. We discuss current and planned uses of consumer-satisfaction information in assuring LTC quality. We end by describing some of the key opportunities for broader use of consumer-satisfaction information and accompanying challenges to be addressed to make more effective use of this information in the effort to improve LTC quality. PMID- 17409047 TI - The development of a CAHPS instrument for Nursing Home Residents (NHCAHPS). AB - We report on a federal initiative to develop a CAHPS (The Consumer Assessment of Healthcare Providers and Systems) survey to measure residents' experiences with quality-of-care and quality-of-life in nursing homes (known as NHCAHPS). We focus on how we created and tested questions for inclusion in the instrument and tested a possible cognitive screener to determine which residents could participate in a NHCAHPS interview. The major lessons learned were: (1) In contrast to other CAHPS surveys, ratings were more useful than reports because of the difficulty that residents had with summarizing over time and people; (2) consistent with other CAHPS surveys, the 0 to 10 response scale appeared to work well with nursing home residents for many of the quality-of-care questions; however, a different response scale was needed for many of the quality-of-life items; and (3) in contrast with typical survey methodology and other CAHPS surveys where explicit time reference periods are used, a non-specific present reference period in questions seemed to work best. PMID- 17409048 TI - Developing and testing a satisfaction survey for nursing home residents: the Ohio Experience. AB - Input from consumers has become an important part of quality improvement in long term care and for consumer decision-making. This paper documents the development of the Ohio Nursing Home Resident Satisfaction Survey (ONHRSS) through a partnership of state government, research, and industry experts. The instrument was tested and refined through two waves of data--a pretest phase and later with statewide data. Exploratory and confirmatory analyses with statewide data identified eight primary factors along with an underlying, secondary Global Satisfaction factor. Reliability of the domains ranged from .69 to .95. Recommendations for further refinement and testing of the instrument are discussed along with policy and practice implications. PMID- 17409049 TI - Implementation and results of the statewide ONHRSS. AB - In response to increased consumer demand, state governments are publishing more detailed information about the quality of nursing homes. In 2002, Ohio legislation mandated the publication of a Long-Term Care Consumer Guide that included information on nursing home resident and family satisfaction surveys. Using the 2002 Ohio Nursing Home Resident Satisfaction Survey as an example, this article addresses two research questions: (1) how can statewide resident face-to face satisfaction interviews be implemented to maximize participation of residents and facilities and produce reliable and valid results, and (2) how satisfied are residents with their nursing home experiences, overall and by level of cognitive function. PMID- 17409050 TI - The relationship between organizational factors and resident satisfaction with nursing home care and life. AB - We examined the relationships between nursing home (NH) resident satisfaction and NH organizational characteristics, while controlling for the effect of resident characteristics within facilities. We used a stratified, random sample of NHs (N = 72) from two states and a prescreened and randomized sample of 1496 residents. Data sources included resident interviews, an administrator survey, the Minimum Data Set (MDS), and the Online Survey, Certification and Reporting System (OSCAR). Using Hierarchical Linear Modeling (HLM) techniques, we found that non chain affiliation, certified nursing assistant staffing, and provision of a family council had significant positive effects on total resident satisfaction. The presence of a special care unit was associated with lower levels of satisfaction. PMID- 17409052 TI - Temporal disintegration in depersonalization disorder. AB - Distortions of the experience of time are central to some types of dissociative experiences. In this study, we investigated the relationship between a self report measure of temporal disintegration and symptoms of dissociation in depersonalization disorder (DPD). Fifty-two DPD and thirty non-clinical control participants were administered the Dissociative Experience Scale (DES) and Temporal Integration Inventory (TII). The DPD group had significantly higher TII scores than the control group. Within the DPD group, there was a significant positive correlation between DES total score and TII total score, and between TII time distinction subscale score and TII-agency subscale score. In the DPD group, TII scores were not associated with age of onset or duration of illness. Of the three dissociative domains of absorption, amnesia, and depersonalization/derealization, only absorption was a significant predictor of TII total and subscale scores by stepwise linear regression analyses. We conclude that the experience of temporal disintegration in DPD is not directly related to the core symptoms of depersonalization/derealization, but exists when the depersonalized experience involves more prominent absorption. PMID- 17409053 TI - Building a secure internal attachment: an intra-relational approach to ego strengthening and emotional processing with chronically traumatized clients. AB - In this paper, we introduce Intra-Relational AEDP (I-R) as an attachment-based experiential approach to trauma treatment. Integrating Accelerated Experiential Dynamic Psychotherapy (AEDP) (Fosha, 2000a,b, 2002, 2003) with ego-state methodology, intra-relational interventions specifically seeks to help clients by (1) fostering capacities for self-regulation through shared states of affective resonance between therapist, client, and dissociated self-states; (2) facilitating authentic, open internal dialogue between self-states which can alter engrained patterns of intra-psychic conflict and self-punishment; (3) developing abilities for self-reflection and emotional processing by co-mingling previously disowned affect and emotional memories with here and now experience; and (4) attending to positive affects evoked through experiences of transformation, self-compassion, and self-affirmation. Drawing from object relations and attachment theory, intra-relational interventions places particular emphasis on internal attachment relationships formed through interactions between the client's subjective selves (internal subjects) and reflective selves (internal objects). Through visual imagery, internal dialogue, and explicit relational techniques, intra-relational interventions aims to develop this subjective-reflective dyad's capacity for reciprocal attunement, resonance, and responsiveness. Such clinical strategies aim to foster healing and psychological integration between the client and heretofore disavowed aspects of self. PMID- 17409054 TI - Is peritraumatic dissociation a risk factor for PTSD? AB - In the literature, peritraumatic dissociation is frequently considered to be a risk factor for Posttraumatic Stress Disorder (PTSD). In the last few years, a large number of studies have investigated the connections between PTSD and peritraumatic dissociation. A meta-analysis was conducted, including 35 empirical studies that discuss the connections between peritraumatic dissociation and PTSD. Meta-analysis makes it possible to undertake a systematic integration of findings produced by primary studies of this kind to date. The average effect size was r=0.36, indicating a significant positive correlation between the two dimensions. In this article, the authors make a distinction between correlate and risk factor. Therefore, subgroup analyses of quasi-prospective and retrospective studies were undertaken with a view to establishing whether peritraumatic dissociation represents a risk factor for the development of PTSD following a traumatic event. The average effect size in the quasi-prospective studies was r=0.34, which was significant. From a methodological viewpoint, the results show that peritraumatic dissociation is a moderate risk factor for PTSD. Nonetheless, the conceptualization of peritraumatic dissociation in the framework of psychotraumatic stress syndromes--is it a predictor, a symptom, or something else entirely?--needs to be addressed by future research. PMID- 17409055 TI - Borderline personality disorder and dissociation. AB - This study examined the dissociative comorbidity reported by psychiatric inpatients with and without borderline personality disorder. The Dissociative Disorders Interview Schedule and Dissociative Experiences Scale were administered to 201 general adult psychiatric inpatients; of these, 110 also received the Structured Clinical Interview for DSM-IV-TR Dissociative Disorders. The patients were then divided into two groups--a group of 93 who met criteria for borderline personality disorder on the Dissociative Disorders Interview Schedule and 108 who did not. The two groups were then compared on dissociative symptoms and disorders. The inpatients with borderline personality disorder reported significantly more dissociative symptoms and disorders on all measures. On the Dissociative Disorders Interview Schedule, 59% of the borderline patients met criteria for a dissociative disorder compared with 22% of the non-borderline patients. Chronic, complex dissociative symptoms and disorders are common in borderline personality disorder. The ninth DSM-IV-TR criterion for borderline personality disorder does not adequately describe the dissociative comorbidity in the disorder. PMID- 17409056 TI - Researching dissociative identity disorder: practical suggestions and ethical implications. AB - This article's purpose is to aid researchers who are interested in conducting research with dissociative populations, but who may not have extensive experience in this area. Researching dissociative identity disorder (DID) comes with many challenges. In this article, the ethics and logistics involved in selecting and implementing research methodology with DID samples are considered. This advice is centered in the author's experience of conducting research with adults in North American hospital and community settings. Researchers in this area should design data collection procedures for maximum flexibility and participant comfort, while maintaining an acceptable level of scientific rigor. Training is essential so that both principal investigators and assistants are able to deal sensitively with trauma survivors. Psychology researchers have a unique ability to be aware of hierarchy and power in the experimental setting, and to consider these issues in order to convey respect for participants. PMID- 17409060 TI - Interphase chromosome arrangement in Arabidopsis thaliana is similar in differentiated and meristematic tissues and shows a transient mirror symmetry after nuclear division. AB - Whole-mount fluorescence in situ hybridization (FISH) was applied to Arabidopsis thaliana seedlings to determine the three-dimensional (3D) interphase chromosome territory (CT) arrangement and heterochromatin location within the positional context of entire tissues or in particular cell types of morphologically well preserved seedlings. The interphase chromosome arrangement was found to be similar between all inspected meristematic and differentiated root and shoot cells, indicating a lack of a gross reorganization during differentiation. The predominantly random CT arrangement (except for a more frequent association of the homologous chromosomes bearing a nucleolus organizer) and the peripheric location of centromeric heterochromatin were as previously observed for flow sorted nuclei, but centromeres tend to fuse more often in nonendoreduplicating cells and NORs in differentiated cells. After mitosis, sister nuclei revealed a symmetric arrangement of homologous CTs waning with the progress of the cell cycle or in the course of differentiation. Thus, the interphase chromosome arrangement in A. thaliana nuclei seems to be constrained mainly by morphological features such as nuclear shape, presence or absence of a nucleolus organizer on chromosomes, nucleolar volume, and/or endopolyploidy level. PMID- 17409061 TI - Nup96-dependent hybrid lethality occurs in a subset of species from the simulans clade of Drosophila. AB - The cross of Drosophila melanogaster females to D. simulans males typically produces lethal F(1) hybrid males. F(1) male lethality is suppressed when the D. simulans Lhr(1) hybrid rescue strain is used. Viability of these F(1) males carrying Lhr(1) is in turn substantially reduced when the hybrids are heterozygous for some mutant alleles of the D. melanogaster Nup96 gene. I show here that similar patterns of Nup96-dependent lethality occur when other hybrid rescue mutations are used to create F(1) males, demonstrating that Nup96 does not reduce hybrid viability by suppressing the Lhr(1) rescue effect. The penetrance of this Nup96-dependent lethality does not correlate with the penetrance of the F(1) hybrid rescue, arguing that these two phenomena reflect genetically independent processes. D. simulans, together with two additional sister species, forms a clade that speciated after the divergence of their common ancestor from D. melanogaster. I report here that Nup96(-) reduces F(1) viability in D. melanogaster hybrids with one of these sister species, D. sechellia, but not with the other, D. mauritiana. These results suggest that Nup96-dependent lethality evolved after the speciation of D. melanogaster from the common ancestor of the simulans clade and is caused by an interaction among Nup96, unknown gene(s) on the D. melanogaster X chromosome, and unknown autosomal gene(s), at least some of which have diverged in D. simulans and D. sechellia but not in D. mauritiana. The genetic properties of Nup96 are also discussed relative to other hybrid lethal genes. PMID- 17409062 TI - Suppression of the Schizosaccharomyces pombe cut12.1 cell-cycle defect by mutations in cdc25 and genes involved in transcriptional and translational control. AB - Cdc25 phosphatase primes entry to mitosis by removing the inhibitory phosphate that is transferred to mitosis promoting factor (MPF) by Wee1 related kinases. A positive feedback loop then boosts Cdc25 and represses Wee1 activities to drive full-scale MPF activation and commitment to mitosis. Dominant mutations in the Schizosaccharomyces pombe spindle pole body (SPB) component Cut12 enable cdc25.22 mutants to overcome a G2 arrest at 36 degrees and enter mitosis. The recessive temperature-sensitive cut12.1 mutation results in the formation of monopolar spindles in which the spindle pole marker Sad1 is enriched on the nonfunctional SPB at 36 degrees . We identified mutations at five loci that suppressed the lethality of the recessive cut12.1 mutation at 36 degrees and conferred lethality at 20 degrees . Three of the five mutations led to the formation of monopolar spindles at restrictive temperatures, affected cell size at commitment to mitosis, and generated multiple Sad1 foci at nuclear periphery. The five loci, tfb2.rt1, tfb5.rt5, pla1.rt3, rpl4301.rt4, and rot2.1, and multicopy suppressors, including tfb1(+) and dbp10(+), are involved in transcription, translation, or RNA processing, prompting us to establish that elevating Cdc25 levels with the dominant cdc25.d1 allele, suppressed cut12.1. Thus, rot mutants provide a further link between protein production and cell-cycle progression. PMID- 17409063 TI - The centromeric retrotransposons of rice are transcribed and differentially processed by RNA interference. AB - Retrotransposons consist of significant portions of many complex eukaryotic genomes and are often enriched in heterochromatin. The centromeric retrotransposon (CR) family in grass species is colonized in the centromeres and highly conserved among species that have been diverged for >50 MY. These unique characteristics have inspired scientists to speculate about the roles of CR elements in organization and function of centromeric chromatin. Here we report that the CRR (CR of rice) elements in rice are highly enriched in chromatin associated with H3K9me2, a hallmark for heterochromatin. CRR elements were transcribed in root, leaf, and panicle tissues, suggesting a constitutive transcription of this retrotransposon family. However, the overall transcription level was low and the CRR transcripts appeared to be derived from relatively few loci. The majority of the CRR transcripts had chimerical structures and contained only partial CRR sequences. We detected small RNAs (smRNAs) cognate to nonautonomous CRR1 (noaCRR1) and CRR1, but not CRR2 elements. This result was also confirmed by in silico analysis of rice smRNA sequences. These results suggest that different CRR subfamilies may play different roles in the RNAi mediated pathway for formation and maintenance of centromeric heterochromatin. PMID- 17409064 TI - Divergent regulatory OsMADS2 functions control size, shape and differentiation of the highly derived rice floret second-whorl organ. AB - Functional diversification of duplicated genes can contribute to the emergence of new organ morphologies. Model eudicot plants like Arabidopsis thaliana and Antirrhinum majus have a single PI/GLO gene that together with AP3/DEF regulate petal and stamen formation. Lodicules of grass flowers are morphologically distinct reduced organs occupying the position of petals in other flowers. They serve a distinct function in partial and transient flower opening to allow stamen emergence and cross-pollination. Grasses have duplicated PI/GLO-like genes and in rice (Oryza sativa) one these genes, OsMADS2, controls lodicule formation without affecting stamen development. In this study, we investigate the mechanistic roles played by OsMADS2. We ascribe a function for OsMADS2 in controlling cell division and differentiation along the proximal-distal axis. OsMADS2 is required to trigger parenchymatous and lodicule-specific vascular development while maintaining a small organ size. Our data implicate the developmentally late spatially restricted accumulation of OsMADS2 transcripts in the differentiating lodicule to control growth of these regions. The global architecture of transcripts regulated by OsMADS2 gives insights into the regulation of cell division and vascular differentiation that together can form this highly modified grass organ with important functions in floret opening and stamen emergence independent of the paralogous gene OsMADS4. PMID- 17409065 TI - Integrative mapping of Gossypium hirsutum L. by meiotic fluorescent in situ hybridization of a tandemly repetitive sequence (B77). AB - We determined the relative positions of the tandem-repeat molecular cytogenetic marker B77, translocation breakpoints, and telosome arms in Gossypium hirsutum cytogenetic stocks by fluorescence in situ hybridization (FISH) analysis of meiotic quadrivalents in 16 single and 2 double translocation heterozygotes and five monotelodisomics. Results delimited the B77 FISH locus to the right arm of the D-subgenome chromosome 14 (14R) and the short arm (14sh), respectively. By equating 14R with 14sh and 14L (left) with 14Lo (long), the findings established a unified nomenclature for the arms of chromosome 14. Previously reported chromosome 14 arm locations were confirmed for four of the five translocations involving chromosome 14, namely NT1L-14L (2780), NT2R-14R (2B-1), NT14L-23R (2777), and NT14R-24R (2781), whereas the location of breakpoint T6L-14L was not confirmed and was reassigned to arm 14R. When used as a probe on Southern blots, the B77 signal was associated with a terminus of the D-subgenome RFLP linkage group (LG) D04 by linkage analysis of an interspecific F(2) population, now known to be chromosome 20. However, additional codominant DNA marker information in the affected region excluded the B77 polymorphism detected by Southern blot hybridization from chromosome 20 and, indeed, from the remainder of the genome. PMID- 17409066 TI - Transcription of the 1.688 satellite DNA family is under the control of RNA interference machinery in Drosophila melanogaster ovaries. AB - Here we show that RNA interference (RNAi) machinery operates in Drosophila melanogaster 1.688 satellite transcription. Mutation in the spn-E gene, known to be involved in RNAi in the oocytes, causes an increase of satellite transcript abundance. Transcripts of both strands of 1.688 satellite repeats in germinal tissues were detected. The strength of the effects of the spn-E mutation differs for 1.688 satellite DNA subfamilies and is more pronounced for autosomal pericentromeric satellites compared to the X-linked centromeric ones. The spn E(1) mutation causes an increase of the H3-AcK9 mark and TAF1 (a component of the polymerase II transcriptional complex) occupancy in the chromatin of autosomal pericentromeric repeats. Thus, we revealed that RNAi operates in ovaries to maintain the silenced state of centromeric and pericentromeric 1.688 repeats. PMID- 17409067 TI - Stepping-stone spatial structure causes slow decay of linkage disequilibrium and shifts the site frequency spectrum. AB - The symmetric island model with D demes and equal migration rates is often chosen for the investigation of the consequences of population subdivision. Here we show that a stepping-stone model has a more pronounced effect on the genealogy of a sample. For samples from a small geographical region commonly used in genetic studies of humans and Drosophila, there is a shift of the frequency spectrum that decreases the number of low-frequency-derived alleles and skews the distribution of statistics of Tajima, Fu and Li, and Fay and Wu. Stepping-stone spatial structure also changes the two-locus sampling distribution and increases both linkage disequilibrium and the probability that two sites are perfectly correlated. This may cause a false prediction of cold spots of recombination and may confuse haplotype tests that compute probabilities on the basis of a homogeneously mixing population. PMID- 17409068 TI - toutvelu, a regulator of heparan sulfate proteoglycan biosynthesis, controls guidance cues for germ-cell migration. AB - The primitive embryonic gonad in Drosophila melanogaster is composed of germ cells and somatic gonadal precursor cells (SGPs). The assembly of a functional gonad involves a complex series of germ-cell migration events, which are thought to be guided by attractive and repulsive cues. Here, we demonstrate a novel role for toutvelu (ttv), a regulator of heparan sulfate proteoglycan biosynthesis during this process. Germline clonal analysis suggests that maternal deposition of ttv is required for proper germ-cell migration. Conversely, ectopic expression of ttv in early embryos results in severe germ-cell migration defects and inappropriate spreading of Hh protein. Moreover, overexpression of ttv in only the receiving cells, rather than in the sending cells, leads to phenotypic consequences. Finally, supporting the claim that the signaling molecule Hedgehog (Hh) may function as a chemoattractant to guide germ cells, errant germ cells are found localized near pockets containing high concentrations of Hh protein. PMID- 17409069 TI - A microsatellite-based, gene-rich linkage map reveals genome structure, function and evolution in Gossypium. AB - The mapping of functional genes plays an important role in studies of genome structure, function, and evolution, as well as allowing gene cloning and marker assisted selection to improve agriculturally important traits. Simple sequence repeats (SSRs) developed from expressed sequence tags (ESTs), EST-SSR (eSSR), can be employed as putative functional marker loci to easily tag corresponding functional genes. In this paper, 2218 eSSRs, 1554 from G. raimondii-derived and 754 from G. hirsutum-derived ESTs, were developed and used to screen polymorphisms to enhance our backbone genetic map in allotetraploid cotton. Of the 1554 G. raimondii-derived eSSRs, 744 eSSRs were able to successfully amplify polymorphisms between our two mapping parents, TM-1 and Hai7124, presenting a polymorphic rate of 47.9%. However, only a 23.9% (159/754) polymorphic rate was produced from G. hirsutum-derived eSSRs. No relationship was observed between the level of polymorphism, motif type, and tissue origin, but the polymorphism appeared to be correlated with repeat type. After integrating these new eSSRs, our enhanced genetic map consists of 1790 loci in 26 linkage groups and covers 3425.8 cM with an average intermarker distance of 1.91 cM. This microsatellite based, gene-rich linkage map contains 71.96% functional marker loci, of which 87.11% are eSSR loci. There were 132 duplicated loci bridging 13 homeologous At/Dt chromosome pairs. Two reciprocal translocations after polyploidization between A2 and A3, and between A4 and A5, chromosomes were further confirmed. A functional analysis of 975 ESTs producing 1122 eSSR loci tagged in the map revealed that 60% had clear BLASTX hits (<1e(-10)) to the Uniprot database and that 475 were associated mainly with genes belonging to the three major gene ontology categories of biological process, cellular component, and molecular function; many of the ESTs were associated with two or more category functions. The results presented here will provide new insights for future investigations of functional and evolutionary genomics, especially those associated with cotton fiber improvement. PMID- 17409070 TI - DDB2, DDB1A and DET1 exhibit complex interactions during Arabidopsis development. AB - Damaged DNA-binding proteins 1 and 2 (DDB1 and DDB2) are subunits of the damaged DNA-binding protein complex (DDB). DDB1 is also found in the same complex as DE ETIOLATED 1 (DET1), a negative regulator of light-mediated responses in plants. Arabidopsis has two DDB1 homologs, DDB1A and DDB1B. ddb1a single mutants have no visible phenotype while ddb1b mutants are lethal. We have identified a partial loss-of-function allele of DDB2. To understand the genetic interaction among DDB2, DDB1A, and DET1 during Arabidopsis light signaling, we generated single, double, and triple mutants. det1 ddb2 partially enhances the short hypocotyl and suppresses the high anthocyanin content of dark-grown det1 and suppresses the low chlorophyll content, early flowering time (days), and small rosette diameter of light-grown det1. No significant differences were observed between det1 ddb1a and det1 ddb1a ddb2 in rosette diameter, dark hypocotyl length, and anthocyanin content, suggesting that these are DDB1A-dependent phenotypes. In contrast, det1 ddb1a ddb2 showed higher chlorophyll content and later flowering time than det1 ddb1a, indicating that these are DDB1A-independent phenotypes. We propose that the DDB1A-dependent phenotypes indicate a competition between DDB2- and DET1 containing complexes for available DDB1A, while, for DDB1A-independent phenotypes, DDB1B is able to fulfill this role. PMID- 17409071 TI - The biologically relevant targets and binding affinity requirements for the function of the yeast actin-binding protein 1 Src-homology 3 domain vary with genetic context. AB - Many protein-protein interaction domains bind to multiple targets. However, little is known about how the interactions of a single domain with many proteins are controlled and modulated under varying cellular conditions. In this study, we investigated the in vivo effects of Abp1p SH3 domain mutants that incrementally reduce target-binding affinity in four different yeast mutant backgrounds in which Abp1p activity is essential for growth. Although the severity of the phenotypic defects observed generally increased as binding affinity was reduced, some genetic backgrounds (prk1 Delta and sla1 Delta) tolerated large affinity reductions while others (sac6 Delta and sla2 Delta) were much more sensitive to these reductions. To elucidate the mechanisms behind these observations, we determined that Ark1p is the most important Abp1p SH3 domain interactor in prk1 Delta cells, but that interactions with multiple targets, including Ark1p and Scp1p, are required in the sac6 Delta background. We establish that the Abp1p SH3 domain makes different, functionally important interactions under different genetic conditions, and these changes in function are reflected by changes in the binding affinity requirement of the domain. These data provide the first evidence of biological relevance for any Abp1p SH3 domain-mediated interaction. We also find that considerable reductions in binding affinity are tolerated by the cell with little effect on growth rate, even when the actin cytoskeletal morphology is significantly perturbed. PMID- 17409072 TI - Inorganic phosphate deprivation causes tRNA nuclear accumulation via retrograde transport in Saccharomyces cerevisiae. AB - Nuclear export of tRNA is an essential eukaryotic function, yet the one known yeast tRNA nuclear exporter, Los1, is nonessential. Moreover recent studies have shown that tRNAs can move retrograde from the cytosol to the nucleus by an undefined process. Therefore, additional gene products involved in tRNA nucleus cytosol dynamics have yet to be identified. Synthetic genetic array (SGA) analysis was employed to identify proteins involved in Los1-independent tRNA transport and in regulating tRNA nucleus-cytosol distribution. These studies uncovered synthetic interactions between los1Delta and pho88Delta involved in inorganic phopsphate uptake. Further analysis revealed that inorganic phosphate deprivation causes transient, temperature-dependent nuclear accumulation of mature cytoplasmic tRNA within nuclei via a Mtr10- and retrograde-dependent pathway, providing a novel connection between tRNA subcellular dynamics and phosphate availability. PMID- 17409074 TI - Genetic improvement of traits affected by interactions among individuals: Sib selection schemes. AB - Livestock populations are usually kept in groups. As a consequence, social interactions among individuals affect productivity, health, and welfare. Current selection methods (individual selection), however, ignore those interactions and yield suboptimal or in some cases even negative responses. In principle, selection between groups instead of individuals offers a solution, but has rarely been adopted in practice for two reasons. First, the relationship between group selection theory and common animal breeding concepts, such as the accuracy of selection, is unclear. Second, application of group selection requires keeping selection candidates in groups, which is often undesirable in practice. This work has two objectives. First, we derive expressions for the accuracy of individual and group selection, which provides a measurement of quality for those methods. Second, we investigate the opportunity to improve traits affected by interactions by using information on relatives kept in family groups, while keeping selection candidates individually. The accuracy of selection based on relatives is shown to be an analogy of the classical expression for traits not affected by interactions. Our results show that selection based on relatives offers good opportunities for effective genetic improvement of traits affected by interactions. PMID- 17409073 TI - LIN-61, one of two Caenorhabditis elegans malignant-brain-tumor-repeat-containing proteins, acts with the DRM and NuRD-like protein complexes in vulval development but not in certain other biological processes. AB - Vulval development in Caenorhabiditis elegans is inhibited by the redundant functions of the synthetic multivulva (synMuv) genes. At least 26 synMuv genes have been identified, many of which appear to act via transcriptional repression. Here we report the molecular identification of the class B synMuv gene lin-61, which encodes a protein composed of four malignant brain tumor (MBT) repeats. MBT repeats, domains of approximately 100 amino acids, have been found in multiple copies in a number of transcriptional repressors, including Polycomb-group proteins. MBT repeats are important for the transcriptional repression mediated by these proteins and in some cases have been shown to bind modified histones. C. elegans contains one other MBT-repeat-containing protein, MBTR-1. We demonstrate that a deletion allele of mbtr-1 does not cause a synMuv phenotype nor does mbtr 1 appear to act redundantly with or in opposition to lin-61. We further show that lin-61 is phenotypically and biochemically distinct from other class B synMuv genes. Our data indicate that while the class B synMuv genes act together to regulate vulval development, lin-61 functions separately from some class B synMuv proteins in other biological processes. PMID- 17409075 TI - Transcriptional interferences in cis natural antisense transcripts of humans and mice. AB - For a significant fraction of mRNAs, their expression is regulated by other RNAs, including cis natural antisense transcripts (cis-NATs) that are complementary mRNAs transcribed from opposite strands of DNA at the same genomic locus. The regulatory mechanism of mRNA expression by cis-NATs is unknown, although a few possible explanations have been proposed. To understand this regulatory mechanism, we conducted a large-scale analysis of the currently available data and examined how the overlapping arrangements of cis-NATs affect their expression level. Here, we show that for both human and mouse the expression level of cis NATs decreases as the length of the overlapping region increases. In particular, the proportions of the highly expressed cis-NATs in all cis-NATs examined were approximately 36 and 47% for human and mouse, respectively, when the overlapping region was <200 bp. However, both proportions decreased to virtually zero when the overlapping regions were >2000 bp in length. Moreover, the distribution of the expression level of cis-NATs changes according to different types of the overlapping pattern of cis-NATs in the genome. These results are consistent with the transcriptional collision model for the regulatory mechanism of gene expression by cis-NATs. PMID- 17409076 TI - A novel type of sequence variation: multiple-nucleotide length polymorphisms discovered in the bovine genome. AB - Three types of sequence variations--single-nucleotide polymorphisms (SNPs), insertions and deletions (indels), and short tandem repeats (STRs)--have been extensively reported in mammalian genomes. In this study, we discovered a novel type of sequence variation, i.e., multiple-nucleotide length polymorphisms (MNLPs) in bovine UCN3 (Urocortin 3) and its receptor CRHR2 (corticotropin releasing hormone receptor 2) genes. Both MNLPs featured involvement of multiple nucleotide length polymorphisms (5-18 bases), low sequence identity, and 1.7- to 11-fold changes in promoter activity between two alleles. Therefore, this novel genetic complexity would contribute significantly to the evolutionary, functional, and phenotypic complexity of genomes within or among species. PMID- 17409077 TI - The balance between the novel protein target of wingless and the Drosophila Rho associated kinase pathway regulates planar cell polarity in the Drosophila wing. AB - Planar cell polarity (PCP) signaling is mediated by the serpentine receptor Frizzled (Fz) and transduced by Dishevelled (Dsh). Wingless (Wg) signaling utilizes Drosophila Frizzled 2 (DFz2) as a receptor and also requires Dsh for transducing signals to regulate cell proliferation and differentiation in many developmental contexts. Distinct pathways are activated downstream of Dsh in Wg- and Fz-signaling pathways. Recently, a number of genes, which have essential roles as downstream components of PCP signaling, have been identified in Drosophila. They include the small GTPase RhoA/Rho1, its downstream effector Drosophila rho-associated kinase (Drok), and a number of genes such as inturned (in) and fuzzy (fy), whose biochemical functions are unclear. RhoA and Drok provide a link from Fz/Dsh signaling to the modulation of actin cytoskeleton. Here we report the identification of the novel gene target of wingless (tow) by enhancer trap screening. tow expression is negatively regulated by Wg signaling in wing imaginal discs, and the balance between tow and the Drok pathway regulates wing-hair morphogenesis. A loss-of-function mutation in tow does not result in a distinct phenotype. Genetic interaction and gain-of-function studies provide evidence that Tow acts downstream of Fz/Dsh and plays a role in restricting the number of hairs that wing cells form. PMID- 17409078 TI - An exact nonparametric method for inferring mosaic structure in sequence triplets. AB - Statistical tests for detecting mosaic structure or recombination among nucleotide sequences usually rely on identifying a pattern or a signal that would be unlikely to appear under clonal reproduction. Dozens of such tests have been described, but many are hampered by long running times, confounding of selection and recombination, and/or inability to isolate the mosaic-producing event. We introduce a test that is exact, nonparametric, rapidly computable, free of the infinite-sites assumption, able to distinguish between recombination and variation in mutation/fixation rates, and able to identify the breakpoints and sequences involved in the mosaic-producing event. Our test considers three sequences at a time: two parent sequences that may have recombined, with one or two breakpoints, to form the third sequence (the child sequence). Excess similarity of the child sequence to a candidate recombinant of the parents is a sign of recombination; we take the maximum value of this excess similarity as our test statistic Delta(m,n,b). We present a method for rapidly calculating the distribution of Delta(m,n,b) and demonstrate that it has comparable power to and a much improved running time over previous methods, especially in detecting recombination in large data sets. PMID- 17409079 TI - basA regulates cell wall organization and asexual/sexual sporulation ratio in Aspergillus nidulans. AB - Sphingolipid C4 hydroxylase catalyzes the conversion of dihydrosphingosine to phytosphingosine. In Saccharomyces cerevisiae, Sur2 is essential for sphingolipid C4 hydroxylation activity but not essential for normal growth. Here we demonstrate that the Aspergillus nidulans Sur2 homolog BasA is also required for phytosphingosine biosynthesis but is also essential for viability. We previously reported that a point missense mutation in basA resulted in aberrant cell wall thickening. Here our data suggest that accumulation of dihydrosphingosine is responsible for this phenotype. In addition, two different mutations in basA consistently accelerated the transition from asexual development to sexual development compared to the wild-type strain. The phenotype could be suppressed by exogenous addition of phytosphingosine. Northern analysis suggests that faster sexual development in the basA mutant might be due to a higher transcription level of ppoA and steA, genes demonstrated to coordinate a balance between asexual and sexual development in A. nidulans. Consistent with these findings, mutations in the ceramide-synthase-encoding genes barA and lagA also caused faster transition from asexual to sexual development, supporting the involvement of sphingolipid metabolism in fungal morphogenesis. PMID- 17409080 TI - Genetic modifiers of Drosophila palmitoyl-protein thioesterase 1-induced degeneration. AB - Infantile neuronal ceroid lipofuscinosis (INCL) is a pediatric neurodegenerative disease caused by mutations in the human CLN1 gene. CLN1 encodes palmitoyl protein thioesterase 1 (PPT1), suggesting an important role for the regulation of palmitoylation in normal neuronal function. To further elucidate Ppt1 function, we performed a gain-of-function modifier screen in Drosophila using a collection of enhancer-promoter transgenic lines to suppress or enhance the degeneration produced by overexpression of Ppt1 in the adult visual system. Modifier genes identified in our screen connect Ppt1 function to synaptic vesicle cycling, endo lysosomal trafficking, synaptic development, and activity-dependent remodeling of the synapse. Furthermore, several homologs of the modifying genes are known to be regulated by palmitoylation in other systems and may be in vivo substrates for Ppt1. Our results complement recent work on mouse Ppt1(-/-) cells that shows a reduction in synaptic vesicle pools in primary neuronal cultures and defects in endosomal trafficking in human fibroblasts. The pathways and processes implicated by our modifier loci shed light on the normal cellular function of Ppt1. A greater understanding of Ppt1 function in these cellular processes will provide valuable insight into the molecular etiology of the neuronal dysfunction underlying the disease. PMID- 17409081 TI - SFP genotyping from affymetrix arrays is robust but largely detects cis-acting expression regulators. AB - The recent development of Affymetrix chips designed from assembled EST sequences has spawned considerable interest in identifying single-feature polymorphisms (SFPs) from transcriptome data. SFPs are valuable genetic markers that potentially offer a physical link to the structural genes themselves. However, most current SFP prediction methodologies were developed for sequenced species although SFPs are particularly valuable for species with complex and unsequenced genomes. To establish the sensitivity and specificity of prediction, we explored four methods for identifying SFPs from experiments involving two tissues in two commercial barleys and their doubled-haploid progeny. The methods were compared in terms of numbers of SFPs predicted and their ability to identify known sequence polymorphisms in the features, to confirm existing SNP genotypes and to match existing maps and individual haplotypes. We identified >4000 separate SFPs that accurately predicted the SNP genotype of >98% of the doubled-haploid (DH) lines. They were highly enriched for features containing sequence polymorphisms but all methods uniformly identified a majority of SFPs ( approximately 64%) in features for which there was no sequence polymorphism while 5% mapped to different locations, indicating that "SFPs" mainly represent polymorphism in cis acting regulators. All methods are efficient and robust at predicting markers for gene mapping. PMID- 17409082 TI - A unified model for functional and statistical epistasis and its application in quantitative trait Loci analysis. AB - Interaction between genes, or epistasis, is found to be common and it is a key concept for understanding adaptation and evolution of natural populations, response to selection in breeding programs, and determination of complex disease. Currently, two independent classes of models are used to study epistasis. Statistical models focus on maintaining desired statistical properties for detection and estimation of genetic effects and for the decomposition of genetic variance using average effects of allele substitutions in populations as parameters. Functional models focus on the evolutionary consequences of the attributes of the genotype-phenotype map using natural effects of allele substitutions as parameters. Here we provide a new, general and unified model framework: the natural and orthogonal interactions (NOIA) model. NOIA implements tools for transforming genetic effects measured in one population to the ones of other populations (e.g., between two experimental designs for QTL) and parameters of statistical and functional epistasis into each other (thus enabling us to obtain functional estimates of QTL), as demonstrated numerically. We develop graphical interpretations of functional and statistical models as regressions of the genotypic values on the gene content, which illustrates the difference between the models--the constraint on the slope of the functional regression--and when the models are equivalent. Furthermore, we use our theoretical foundations to conceptually clarify functional and statistical epistasis, discuss the advantages of NOIA over previous theory, and stress the importance of linking functional and statistical models. PMID- 17409083 TI - Survival quantitative trait locus fine mapping by measuring and testing for Hardy Weinberg and linkage disequilibrium. AB - I show that fine-scale localization of a survival-related locus can be accomplished on the basis of deviations from Hardy-Weinberg equilibrium and linkage disequilibrium at closely linked marker loci. The method is based on chi(2)-tests and they can be performed for age-specific samples of alive (or dead) individuals, as for combined samples of alive and dead individuals. PMID- 17409084 TI - Temporal dynamics and linkage disequilibrium in natural Caenorhabditis elegans populations. AB - Caenorhabditis elegans is a major laboratory model system yet a newcomer to the field of population genetics, and relatively little is known of its biology in the wild. Recent studies of natural populations at a single time point revealed strong spatial population structure and suggested that these populations may be very dynamic. We have therefore studied several natural C. elegans populations over time and genotyped them at polymorphic microsatellite loci. While some populations appear to be genetically stable over the course of observation, others seem to go extinct, with full replacement of multilocus genotypes upon regrowth. The frequency of heterozygotes indicates that outcrossing occurs at a mean frequency of 1.7% and is variable between populations. However, in genetically stable populations, linkage disequilibrium between different chromosomes can be maintained over several years at a level much higher than expected from the heterozygote frequency. C. elegans seems to follow metapopulation dynamics, and the maintenance of linkage disequilibrium despite a low yet significant level of outcrossing suggests that selection may act against the progeny of outcrossings. PMID- 17409085 TI - Adaptation of a quantitative trait to a moving optimum. AB - We investigate adaptive evolution of a quantitative trait under stabilizing selection with a moving optimum. We characterize three regimes, depending on whether (1) the beneficial mutation rate, (2) the fixation time, or (3) the rate of environmental change is the limiting factor for adaptation. If the environment is rate limiting, mutations with a small phenotypic effect are preferred over large mutations, in contrast to standard theory. PMID- 17409086 TI - Thirty-one flavors of Drosophila rab proteins. AB - Rab proteins are small GTPases that play important roles in transport of vesicle cargo and recruitment, association of motor and other proteins with vesicles, and docking and fusion of vesicles at defined locations. In vertebrates, >75 Rab genes have been identified, some of which have been intensively studied for their roles in endosome and synaptic vesicle trafficking. Recent studies of the functions of certain Rab proteins have revealed specific roles in mediating developmental signal transduction. We have begun a systematic genetic study of the 33 Rab genes in Drosophila. Most of the fly proteins are clearly related to specific vertebrate proteins. We report here the creation of a set of transgenic fly lines that allow spatially and temporally regulated expression of Drosophila Rab proteins. We generated fluorescent protein-tagged wild-type, dominant negative, and constitutively active forms of 31 Drosophila Rab proteins. We describe Drosophila Rab expression patterns during embryogenesis, the subcellular localization of some Rab proteins, and comparisons of the localization of wild type, dominant-negative, and constitutively active forms of selected Rab proteins. The high evolutionary conservation and low redundancy of Drosophila Rab proteins make these transgenic lines a useful tool kit for investigating Rab functions in vivo. PMID- 17409087 TI - Involving undergraduates in the annotation and analysis of global gene expression studies: creation of a maize shoot apical meristem expression database. AB - Through a multi-university and interdisciplinary project we have involved undergraduate biology and computer science research students in the functional annotation of maize genes and the analysis of their microarray expression patterns. We have created a database to house the results of our functional annotation of >4400 genes identified as being differentially regulated in the maize shoot apical meristem (SAM). This database is located at http://sam.truman.edu and is now available for public use. The undergraduate students involved in constructing this unique SAM database received hands-on training in an intellectually challenging environment, which has prepared them for graduate and professional careers in biological sciences. We describe our experiences with this project as a model for effective research-based teaching of undergraduate biology and computer science students, as well as for a rich professional development experience for faculty at predominantly undergraduate institutions. PMID- 17409088 TI - Genomewide association analysis in diverse inbred mice: power and population structure. AB - The discovery of quantitative trait loci (QTL) in model organisms has relied heavily on the ability to perform controlled breeding to generate genotypic and phenotypic diversity. Recently, we and others have demonstrated the use of an existing set of diverse inbred mice (referred to here as the mouse diversity panel, MDP) as a QTL mapping population. The use of the MDP population has many advantages relative to traditional F(2) mapping populations, including increased phenotypic diversity, a higher recombination frequency, and the ability to collect genotype and phenotype data in community databases. However, these methods are complicated by population structure inherent in the MDP and the lack of an analytical framework to assess statistical power. To address these issues, we measured gene expression levels in hypothalamus across the MDP. We then mapped these phenotypes as quantitative traits with our association algorithm, resulting in a large set of expression QTL (eQTL). We utilized these eQTL, and specifically cis-eQTL, to develop a novel nonparametric method for association analysis in structured populations like the MDP. These eQTL data confirmed that the MDP is a suitable mapping population for QTL discovery and that eQTL results can serve as a gold standard for relative measures of statistical power. PMID- 17409089 TI - Variance of the parental genome contribution to inbred lines derived from biparental crosses. AB - The expectation of the parental genome contribution to inbred lines derived from biparental crosses or backcrosses is well known, but no theoretical results exist for its variance. Our objective was to derive the variance of the parental genome contribution to inbred lines developed by the single-seed descent or double haploid method from biparental crosses or backcrosses. We derived formulas and tabulated results for the variance of the parental genome contribution depending on the chromosome lengths and the mating scheme used for inbred line development. A normal approximation of the probability distribution function of the parental genome contribution fitted well the exact distribution obtained from computer simulations. We determined upper and lower quantiles of the parental genome contribution for model genomes of sugar beet, maize, and wheat using normal approximations. These can be employed to detect essentially derived varieties in the context of plant variety protection. Furthermore, we outlined the application of our results to predict the response to selection. Our results on the variance of the parental genome contribution can assist breeders and geneticists in the design of experiments or breeding programs by assessing the variation around the mean parental genome contribution for alternative crossing schemes. PMID- 17409090 TI - High frequency of mutations that expand the host range of an RNA virus. AB - The ability of a virus population to colonize a novel host is predicted to depend on the equilibrium frequency of potential colonists (i.e., genotypes capable of infecting the novel host) in the source population. In this study, we investigated the determinants of the equilibrium frequency of potential colonists in the RNA bacteriophage 6. We isolated 40 spontaneous mutants capable of infecting a novel Pseudomonas syringae host and sequenced their host attachment genes to identify the responsible mutations. We observed 16 different mutations in the host attachment gene and used a new statistical approach to estimate that 39 additional mutations were missed by our screen. Phenotypic and fitness assays confirmed that the proximate mechanism underlying host range expansion was an increase in the ability to attach to the novel host and that acquisition of this ability most often imposed a cost for growth rate on two standard hosts. Considered in a population genetic framework, our data suggest that host range mutations should exist in phage populations at an equilibrium frequency (3 x 10( 4)) that exceeds the phage mutation rate by more than two orders of magnitude. Thus, colonization of novel hosts is unlikely to be limited by an inability to produce appropriate mutations. PMID- 17409091 TI - The genetic basis of zinc tolerance in the metallophyte Arabidopsis halleri ssp. halleri (Brassicaceae): an analysis of quantitative trait loci. AB - The species Arabidopsis halleri, an emerging model for the study of heavy metal tolerance and accumulation in plants, has evolved a high level of constitutive zinc tolerance. Mapping of quantitative trait loci (QTL) was used to investigate the genetic architecture of zinc tolerance in this species. A first-generation backcross progeny of A. halleri ssp. halleri from a highly contaminated industrial site and its nontolerant relative A. lyrata ssp. petraea was produced and used for QTL mapping of zinc tolerance. A genetic map covering most of the A. halleri genome was constructed using 85 markers. Among these markers, 65 were anchored in A. thaliana and revealed high synteny with other Arabidopsis genomes. Three QTL of comparable magnitude on three different linkage groups were identified. At all QTL positions zinc tolerance was enhanced by A. halleri alleles, indicating directional selection for higher zinc tolerance in this species. The two-LOD support intervals associated with these QTL cover 24, 4, and 13 cM. The importance of each of these three regions is emphasized by their colocalization with HMA4, MTP1-A, and MTP1-B, respectively, three genes well known to be involved in metal homeostasis and tolerance in plants. PMID- 17409092 TI - Control of sigma virus multiplication by the ref(2)P gene of Drosophila melanogaster: an in vivo study of the PB1 domain of Ref(2)P. AB - Ref(2)P has been described as one of the Drosophila proteins that interacts with the sigma virus cycle. We generated alleles to identify critical residues involved in the restrictive (inhibiting viral multiplication) or permissive (allowing viral multiplication) character of Ref(2)P. We demonstrate that permissive alleles increase the ability of the sigma virus to infect Drosophila when compared to null alleles and we confirm that restrictive alleles decrease this capacity. Moreover, we have created alleles unfunctional in viral cycling while functional for Ref(2)P fly functions. This type of allele had never been observed before and shows that fly- and virus-related activities of Ref(2)P are separable. The viral status of Ref(2)P variants is determined by the amino terminal PB1 domain polymorphism. In addition, an isolated PB1 domain mimics virus-related functions even if it is similar to a loss of function toward fly related activities. The evolutionary tree of the Ref(2)P PB1 domain that we could build on the basis of the natural allele sequences is in agreement with an evolution of PB1 domain due to successive transient selection waves. PMID- 17409093 TI - Evolution of coadaptation in a subdivided population. AB - The interplay between population subdivision and epistasis is investigated by studying the fixation probability of a coadapted haplotype in a subdivided population. Analytical and simulation models are developed to study the evolutionary fate of two conditionally neutral mutations that interact epistatically to enhance fitness. We find that the fixation probability of a coadapted haplotype shows a marked increase when the population is genetically subdivided and subpopulations are loosely connected by migration. Moderate migration and isolation allow the propagation of the mutant alleles across subpopulations, while at the same time preserving the favorable allelic combination established within each subpopulation. Together they create the condition most favorable for the ultimate fixation of the coadapted haplotype. On the basis of the analytical and simulation results, we discuss the fundamental role of population subdivision and restricted gene flow in promoting the evolution of functionally integrated systems, with some implications for the shifting-balance theory of evolution. PMID- 17409094 TI - Evolution of DNA double-strand break repair by gene conversion: coevolution between a phage and a restriction-modification system. AB - The necessity to repair genome damage has been considered to be an immediate factor responsible for the origin of sex. Indeed, attack by a cellular restriction enzyme of invading DNA from several bacteriophages initiates recombinational repair by gene conversion if there is homologous DNA. In this work, we modeled the interaction between a bacteriophage and a bacterium carrying a restriction enzyme as antagonistic coevolution. We assume a locus on the bacteriophage genome has either a restriction-sensitive or a restriction resistant allele, and another locus determines whether it is recombination/repair proficient or defective. A restriction break can be repaired by a co-infecting phage genome if one of them is recombination/repair proficient. We define the fitness of phage (resistant/sensitive and repair-positive/-negative) genotypes and bacterial (restriction-positive/-negative) genotypes by assuming random encounter of the genotypes, with given probabilities of single and double infections, and the costs of resistance, repair, and restriction. Our results show the evolution of the repair allele depends on b(1)/b(0), the ratio of the burst size b(1) under damage to host cell physiology induced by an unrepaired double-strand break to the default burst size b(0). It was not until this effect was taken into account that the evolutionary advantage of DNA repair became apparent. PMID- 17409095 TI - Structural and biophysical coupling of heparin and activin binding to follistatin isoform functions. AB - Follistatin (FS) regulates transforming growth factor-beta superfamily ligands and is necessary for normal embryonic and ovarian follicle development. Follistatin is expressed as two splice variants (FS288 and FS315). Previous studies indicated differences in heparin binding between FS288 and FS315, potentially influencing the physiological functions and locations of these isoforms. We have determined the structure of the FS315-activin A complex and quantitatively compared heparin binding by the two isoforms. The FS315 complex structure shows that both isoforms inhibit activin similarly, but FS315 exhibits movements within follistatin domain 3 (FSD3) apparently linked to binding of the C-terminal extension. Surprisingly, the binding affinities of FS288 and FS315 for heparin are similar at lower ionic strengths with FS315 binding decreasing more sharply as a function of salt concentration. When bound to activin, FS315 binds heparin similarly to the FS288 isoform, consistent with the structure of the complex, in which the acidic residues of the C-terminal extension cannot interact with the heparin-binding site. Activin-induced binding of heparin is unique to the FS315 isoform and may stimulate clearance of FS315 complexes. PMID- 17409096 TI - Enhanced apoA-I-dependent cholesterol efflux by ABCA1 from sphingomyelin deficient Chinese hamster ovary cells. AB - ATP binding cassette protein A1 (ABCA1) plays a major role in cholesterol homeostasis and high density lipoprotein (HDL) metabolism. It is proposed that ABCA1 reorganizes the plasma membrane and generates more loosely packed domains that facilitate apoA-I-dependent cholesterol efflux. In this study, we examined the effects of the cellular sphingomyelin level on HDL formation by ABCA1 by using a Chinese hamster ovary-K1 mutant cell line, LY-A, which has a missense mutation in the ceramide transfer protein CERT. When LY-A cells were cultured in Nutridoma-BO medium and sphingomyelin content was reduced, apoA-I-dependent cholesterol efflux by ABCA1 from LY-A cells increased 1.65-fold compared with that from LY-A/CERT cells stably transfected with human CERT cDNA. Exogenously added sphingomyelin significantly reduced the apoA-I-dependent efflux of cholesterol from LY-A cells, confirming that the decrease in sphingomyelin content in the plasma membrane stimulates cholesterol efflux by ABCA1. The amount of cholesterol available to cold methyl-beta-cyclodextrin (MbetaCD) extraction from LY-A cells was increased by 40% by the expression of ABCA1 and was 1.6-fold higher than that from LY-A/CERT cells. This step in ABCA1 function, making cholesterol available to cold MbetaCD, was independent of apoA-I. These results suggest that the function of ABCA1 could be divided into two steps: (i) a flopping step to move phosphatidylcholine and cholesterol from the inner to outer leaflet of the plasma membrane, where cholesterol becomes available to cold MbetaCD extraction, and (ii) a loading step to load phosphatidylcholine and cholesterol onto apoA-I to generate HDL. PMID- 17409097 TI - A Bax/Bak-independent mechanism of cytochrome c release. AB - Bax and Bak are multidomain pro-apoptotic members of the Bcl-2 family of proteins that regulate mitochondria-mediated apoptosis by direct modulation of mitochondrial membrane permeability. Since double-knock-out mouse embryonic fibroblasts with deficiency of Bax and Bak are resistant to multiple apoptotic stimuli, Bax and Bak are considered to be an essential gateway for various apoptotic signals. Here we showed that the combination of calcium ionophore A23187 and arachidonic acid induced cytochrome c release and caspase-dependent death of double-knock-out mouse embryonic fibroblasts, indicating that other mechanisms of cytochrome c release exist. Furthermore, A23187/arachidonic acid (ArA)-induced caspase-dependent death was significantly suppressed by the treatment of several serine protease inhibitors including 4-(2 aminoethyl)benzenesulfonylfluoride and l-1-chloro-3-(4-tosylamido)-4-phenyl-2 butanone but not the overexpression of anti-apoptotic Bcl-2 family of proteins or the inhibition of mitochondrial membrane permeability transition. These results indicate that there are at least two mechanisms of cytochrome c release leading to caspase activation, a Bax/Bak-dependent mechanism and a Bax/Bak-independent, but serine protease(s)-dependent, mechanism. PMID- 17409098 TI - Substrate recognition and ubiquitination of SCFSkp2/Cks1 ubiquitin-protein isopeptide ligase. AB - p27, an important cell cycle regulator, blocks the G(1)/S transition in cells by binding and inhibiting Cdk2/cyclin A and Cdk2/cyclin E complexes (Cdk2/E). Ubiquitination and subsequent degradation play a critical role in regulating the levels of p27 during cell cycle progression. Here we provide evidence suggesting that both Cdk2/E and phosphorylation of Thr(187) on p27 are essential for the recognition of p27 by the SCF(Skp2/Cks1) complex, the ubiquitin-protein isopeptide ligase (E3). Cdk2/E provides a high affinity binding site, whereas the phosphorylated Thr(187) provides a low affinity binding site for the Skp2/Cks1 complex. Furthermore, binding of phosphorylated p27/Cdk2/E to the E3 complex showed positive cooperativity. Consistently, p27 is also ubiquitinated in a similarly cooperative manner. In the absence of p27, Cdk2/E and Cks1 increase Skp2 phosphorylation. This phosphorylation enhances Skp2 auto-ubiquitination, whereas p27 inhibits both phosphorylation and auto-ubiquitination of Skp2. PMID- 17409099 TI - Activating transcription factor-1-mediated hepatocyte growth factor-induced down regulation of thrombospondin-1 expression leads to thyroid cancer cell invasion. AB - Hepatocyte growth factor (HGF) plays a major role in the pathogenesis of a variety of human epithelial tumors including papillary carcinoma of the thyroid. Previous reports demonstrated that HGF, acting through the Met receptor, repressed thrombospondin-1 (TSP-1) expression. To study the mechanisms by which HGF down-regulated TSP-1 expression, we transiently transfected a panel of deleted human TSP-1 promoter reporter plasmids into papillary thyroid carcinoma cells. We identified a region between -1210 and -1123 bp relative to the transcription start site that is responsive to HGF treatment and harbors a cAMP responsive element (CRE) at position -1199 (TGACGTCC). Overexpression of various members of the CRE-binding protein family identified activating transcription factor-1 (ATF-1) as the transcription factor responsible for HGF-induced repression of TSP-1 promoter activity. This inhibition was associated with a concomitant increase in the abundance of nuclear ATF-1 protein. Gel shift and antibody supershift studies indicated that ATF-1 was involved in DNA binding to the TSP-1-CRE site. Finally, we utilized small hairpin RNA to target ATF-1 and showed that these small interfering RNA constructs significantly inhibited ATF-1 expression at both the RNA and the protein level. ATF-1 knockdown prevented HGF induced down-regulation of TSP-1 promoter activity and protein expression and also reduced HGF-dependent tumor cell invasion. Taken together, our results indicate that HGF-induced down-regulation of TSP-1 expression is mediated by the interaction of ATF-1 with the CRE binding site in the TSP-1 promoter and that this transcription factor plays a crucial role for tumor invasiveness in papillary carcinoma of the thyroid triggered by HGF. PMID- 17409100 TI - Presence of triploids among oak species. AB - BACKGROUND AND AIMS: Polyploids, organisms that have multiple sets of chromosomes, are common in certain plant and animal taxa. However, there are only a few reports of intraspecific ploidy variation within the genus Quercus. The aim of the study was to investigate the suspected ploidy level of two oaks that have unusual microsatellite banding patterns. METHODS: Polyploidy was investigated by using microsatellite analysis, stomata length measurements and nuclear DNA content estimation by flow cytometry. KEY RESULTS: Each putative triploid tree has patterns of microsatellite variation unexpected for diploid genomes, with up to three alleles at some loci, significantly longer stomata and 1.5 times more DNA per nucleus compared with diploids. CONCLUSIONS: To our knowledge, this report contains the first evidence for triploidy in Q. petraea and confirmation of this phenomenon in Q. robur. Regardless of the positive or negative aspects of the presence of triploid oaks in forest stands, it is of value to be able to screen for them. This study demonstrates that nuclear microsatellites and estimation of DNA content by flow cytometry can readily be used for this purpose. PMID- 17409101 TI - Pandemic H5N1 influenza. PMID- 17409102 TI - Electric breast pump use increases maternal milk volume in African nurseries. AB - Health experts worldwide recognize breastmilk as the superior infant food. Recommendations from the American Academy of Pediatrics and the World Health Organization (WHO) identify exclusive breastfeeding for the first half-year of life and continuation of breastfeeding into toddlerhood as offering maximum protection from illness, providing a substrate for immunological protection. Data from developed countries identify increasing morbidity and mortality rates for infants who have never received breastmilk in life and demonstrate that infants benefit from exclusive breastfeeding, especially, in areas of severe poverty. Preterm infants, most at risk for morbidity and mortality in developing countries, are identified as needing their own mother's milk for survival. Exclusive feeding of own mothers' milk (OMM) is associated with improved infant survival; however, inadequate maternal milk volume (MMV) often necessitates adding artificial feedings or exogenous substances to OMM. The objective of this study was to compare mean daily MMV for mothers of premature or sick infants in special care nurseries (SCN) using one of three methods of OMM expression: electric breast pump, non-electric pedal breast pump, and hand (manual) expression. We studied 65 mothers whose infants were cared for in two SCN in Africa (Kenya and Nigeria) and were unable to feed directly at the breast. In this randomized trial, mothers were randomly assigned to one of three milk expression groups at birth. MMV, the dependent variable, was measured for an average of 8.7 days. MMV for the electric and pedal pump and hand milk expression was 578 +/- 228 ml (n = 22), 463 +/- 302 ml (n = 24) and 323 +/- 199 ml (n = 19), respectively. Data were evaluated using a one-way ANOVA (p = 0.014). The Tukey revealed significant differences (p < 0.01) between electric breast pump expression and hand expression but not between the electric and pedal pump or the pedal pump and hand expression. Findings revealed greater MMV with electric breast pumps than hand-expression for mothers of infants in African nurseries. This data has important implications for international policy if exclusive OMM feeding is to be achieved for the vulnerable infant. Funded by West Virginia University Department of Research and Graduate Studies HSC Grant # 2U023U; Non monetary donations of breast pumps and breast pump kits were made by Medela (Medela, Inc., McHenry, IL, USA). PMID- 17409105 TI - Coronary plaque composition of culprit/target lesions according to the clinical presentation: comment. PMID- 17409106 TI - Prominent posterior mitral annular calcification causing embolic stroke and mimicking left atrial fibroma. PMID- 17409107 TI - Release of cardiac bio-markers during high mechanical index contrast-enhanced echocardiography in humans. AB - BACKGROUND: Recent experimental data have shown that the combined exposure of rodent hearts to high acoustic pressure and ultrasound contrast agents can induce vascular injury and cell damage. The aim of the present work was to test whether similar effects can be observed in humans. METHODS AND RESULTS: Twenty patients underwent simultaneous arterial and coronary sinus blood sampling during contrast enhanced echocardiography using Perfluorocarbon-enhanced Sonicated Dextrose Albumin. Control subjects were compared to groups of patients exposed to either high mechanical index (MI = 1.5) triggered second harmonic (1.3-2.6 MHz) imaging or low mechanical index (MI = 0.2) real-time power modulation imaging for 15 min. No significant changes arterio-venous differences in lactate, total creatine kinase (CK) and myoglobin occurred over time in the three groups. Similarly, the arterio-venous difference in CK-MB and troponin I remained stable over time in control and low-MI patients. By contrast, these two parameters progressively increased over time in the high-MI group (P < 0.05 vs. baseline and vs. controls). CONCLUSION: Our data suggest that high-MI contrast-enhanced echocardiography can cause subclinical release of cardiac bio-markers in humans, while low-MI real-time imaging appears to be safer. PMID- 17409108 TI - Monitoring of therapeutic effect in heart failure patients: a clinical application of 123I MIBG imaging? PMID- 17409109 TI - Evaluation of cardiac sympathetic nerve activity and left ventricular remodelling in patients with dilated cardiomyopathy on the treatment containing carvedilol. AB - AIMS: It has been reported that carvedilol improves cardiac sympathetic nerve activity (CSNA) in patients with dilated cardiomyopathy (DCM). However, the influence of carvedilol on cardiac (123)I-meta-iodobenzylguanidine (MIBG) scintigraphic findings and left ventricular (LV) remodelling has not been determined in DCM patients. METHODS AND RESULTS: In 30 patients with DCM and 10 normal controls, the delayed heart/mediastinum count (H/M) ratio, delayed total defect score (TDS), and washout rate (WR) were determined by (123)I-MIBG scintigraphy. In addition, the left ventricular end-diastolic volume (LVEDV), LV end-systolic volume (LVESV), and LV ejection fraction (LVEF) were calculated by echocardiography. In the DCM patients, the regional defect score index (RDSI), regional washout rate index (RWRI), and wall motion score index (WMSI) were also determined to evaluate regional adrenergic dysfunction and wall motion. Examinations were repeated in all DCM patients after standard treatment containing carvedilol at a dose of 10-20 mg/day (mean dose: 16 +/- 4 mg/day) for a mean of 12 +/- 1 months. Both the (123)I-MIBG scintigraphic and echocardiographic parameters were significantly worse in the DCM patients than the normal control subjects. After treatment, all of these parameters improved significantly in the DCM patients. There was a significant correlation between the changes of (123)I-MIBG findings and changes of the LVEDV and LVESV after treatment. Moreover, there was a significant correlation between changes of the WMSI and those of the RDSI or RWRI in DCM patients. CONCLUSION: Both (123)I-MIBG scintigraphic parameters and echocardiographic parameters were improved in the DCM patients. There was a significant correlation between the changes of (123)I MIBG scintigraphic and echocardiographic findings after treatment. These findings implicate that long-term, including carvedilol, therapy can improve both CSNA and LV remodelling in patients with DCM. PMID- 17409110 TI - Coronary artery perforation during primary PCI: an easily resolved case for a dramatic complication. PMID- 17409111 TI - High plasma cholesteryl ester transfer protein levels may favour reduced incidence of cardiovascular events in men with low triglycerides. AB - AIMS: High cholesteryl ester transfer protein (CETP) concentrations are associated with increased risk of cardiovascular disease (CVD) in subjects with high triglycerides. We determined the relationship of plasma CETP with incident CVD in a population with relatively low triglycerides. METHODS AND RESULTS: A nested case-control study was performed in men participating in the prospective PREVEND study, after exclusion of CVD, diabetes mellitus, and lipid-lowering drugs use at baseline. Plasma CETP was measured in 111 men who developed a cardiovascular event (cases) during follow-up and in 116 controls who remained free of CVD. Fasting total cholesterol (P < 0.001) and triglycerides (P < 0.001) were higher, HDL cholesterol was lower (P = 0.001), but CETP was similar in cases and controls (P = 0.39). Cox proportional hazards regression analysis showed that CVD risk tended to be lower with higher plasma CETP after adjustment for age and lipids (hazard ratio 0.84; 95% CI 0.69-1.03, P = 0.10). Plasma CETP was lower in cases than in controls (P = 0.05) with triglycerides < or = 1.38 mmol/L (median), but not with higher triglycerides. The age-adjusted hazard ratio for CVD was 0.46 (95% CI 0.24-0.90) in men with triglycerides < or = 1.38 mmol/L and CETP > 2.26 mg/L (median) compared with men with similarly low triglycerides and CETP < or = 2.26 mg/L. With higher triglycerides, the hazard ratio for CVD was similar in both CETP categories. CONCLUSION: Relatively high plasma CETP may favour reduced CVD risk in the context of low triglycerides. PMID- 17409112 TI - The proneural determinant MASH1 regulates forebrain Dlx1/2 expression through the I12b intergenic enhancer. AB - Establishment of neuronal networks is an extremely complex process involving the interaction of a diversity of neuronal cells. During mammalian development, these highly organized networks are formed through the differentiation of multipotent neuronal progenitors into multiple neuronal cell lineages. In the developing forebrain of mammals, the combined function of the Dlx1, Dlx2, Dlx5 and Dlx6 homeobox genes is necessary for the differentiation of the GABAergic interneurons born in the ventricular and subventricular zones of the ventral telencephalon, as well as for the migration of these neurons to the hippocampus, cerebral cortex and olfactory bulbs. The 437 bp I12b enhancer sequence in the intergenic region of the Dlx1/2 bigene cluster is involved in the forebrain regulation of Dlx1/2. Using DNase I footprinting, we identified six regions of I12b potentially bound by transcription factors. Mutagenesis of each binding site affected the expression of reporter constructs in transgenic mice. However, the effects of impairing protein-DNA interactions were not uniform across the forebrain Dlx1/2 expression domains, suggesting that distinct regulatory interactions are taking place in the different populations of neuronal precursors. Analyses of protein DNA interactions provide evidence of a direct role for MASH1 in Dlx1/2 regulation in the forebrain. DLX proteins play a crucial role in the maintenance of their own expression, as shown by transgenic and co-transfection experiments. These studies suggest that the seemingly continuous domains of Dlx gene expression in the telencephalon and diencephalon are in fact the combination of distinct cell populations within which different genetic regulatory interactions take place. PMID- 17409113 TI - tailup, a LIM-HD gene, and Iro-C cooperate in Drosophila dorsal mesothorax specification. AB - The LIM-HD gene tailup (tup; also known as islet) has been categorised as a prepattern gene that antagonises the formation of sensory bristles on the notum of Drosophila by downregulating the expression of the proneural achaete-scute genes. Here we show that tup has an earlier function in the development of the imaginal wing disc; namely, the specification of the notum territory. Absence of tup function causes cells of this anlage to upregulate different wing-hinge genes and to lose expression of some notum genes. Consistently, these cells differentiate hinge structures or modified notum cuticle. The LIM-HD co-factors Chip and Ssdp are also necessary for notum specification. This suggests that Tup acts in this process in a complex with Chip and Ssdp. Overexpression of tup, together with araucan, a 'pronotum' gene of the iroquois complex (Iro-C), synergistically reinforces the weak capacity of either gene, when overexpressed singly, to induce ectopic notum-like development. Whereas the Iro-C genes are activated in the notum anlage by EGFR signalling, tup is positively regulated by Dpp signalling. Our data support a model in which the EGFR and Dpp signalling pathways, with their respective downstream Iro-C and tup genes, converge and cooperate to commit cells to the notum developmental fate. PMID- 17409114 TI - The Mrj co-chaperone mediates keratin turnover and prevents the formation of toxic inclusion bodies in trophoblast cells of the placenta. AB - Defects in protein-folding and -degradation machinery have been identified as a major cause of intracellular protein aggregation and of aggregation-associated diseases. In general, it remains unclear how these aggregates are harmful to normal cellular function. We demonstrate here that, in the developing placenta of the mouse, the absence of the Mrj (Dnajb6) co-chaperone prevents proteasome degradation of keratin 18 (K18; Krt18) intermediate filaments, resulting in the formation of keratin inclusion bodies. These inclusions in chorionic trophoblast cells prevent chorioallantoic attachment during placental development. We show further that keratin-deficient embryos undergo chorioallantoic attachment and that, by genetically reducing keratin expression in Mrj(-/-) conceptuses, chorioallantoic attachment was rescued. Therefore, the chorioallantoic attachment phenotype in Mrj mutants is not due to a deficiency of the normal keratin cytoskeleton, but rather is cytotoxicity caused by keratin aggregates that disrupt chorion trophoblast cell organization and function. PMID- 17409115 TI - The F-actin-microtubule crosslinker Shot is a platform for Krasavietz-mediated translational regulation of midline axon repulsion. AB - Axon extension and guidance require a coordinated assembly of F-actin and microtubules as well as regulated translation. The molecular basis of how the translation of mRNAs encoding guidance proteins could be closely tied to the pace of cytoskeletal assembly is poorly understood. Previous studies have shown that the F-actin-microtubule crosslinker Short stop (Shot) is required for motor and sensory axon extension in the Drosophila embryo. Here, we provide biochemical and genetic evidence that Shot functions with a novel translation inhibitor, Krasavietz (Kra, Exba), to steer longitudinally directed CNS axons away from the midline. Kra binds directly to the C-terminus of Shot, and this interaction is required for the activity of Shot to support midline axon repulsion. shot and kra mutations lead to weak robo-like phenotypes, and synergistically affect midline avoidance of CNS axons. We also show that shot and kra dominantly enhance the frequency of midline crossovers in embryos heterozygous for slit or robo, and that in kra mutant embryos, some Robo-positive axons ectopically cross the midline that normally expresses the repellent Slit. Finally, we demonstrate that Kra also interacts with the translation initiation factor eIF2beta and inhibits translation in vitro. Together, these data suggest that Kra-mediated translational regulation plays important roles in midline axon repulsion and that Shot functions as a direct physical link between translational regulation and cytoskeleton reorganization. PMID- 17409116 TI - Protease nexin 1 and its receptor LRP modulate SHH signalling during cerebellar development. AB - Development of the postnatal cerebellum relies on the tight regulation of cell number by morphogens that control the balance between cell proliferation, survival and differentiation. Here, we analyze the role of the serine-protease inhibitor protease nexin 1 (PN-1; SERPINE2) in the proliferation and differentiation of cerebellar granular neuron precursors (CGNPs) via the modulation of their main mitogenic factor, sonic hedgehog (SHH). Our studies show that PN-1 interacts with low-density lipoprotein receptor-related proteins (LRPs) to antagonize SHH-induced CGNP proliferation and that it inhibits the activity of the SHH transcriptional target GLI1. The binding of PN-1 to LRPs interferes with SHH-induced cyclin D1 expression. CGNPs isolated from Pn-1-deficient mice exhibit enhanced basal proliferation rates due to overactivation of the SHH pathway and show higher sensitivity to exogenous SHH. In vivo, the Pn-1 deficiency alters the expression of SHH target genes. In addition, the onset of CGNP differentiation is delayed, which results in an enlarged outer external granular layer. Furthermore, the Pn-1 deficiency leads to an overproduction of CGNPs and to enlargement of the internal granular layer in a subset of cerebellar lobes during late development and adulthood. We propose that PN-1 contributes to shaping the cerebellum by promoting cell cycle exit. PMID- 17409117 TI - grp (chk1) replication-checkpoint mutations and DNA damage trigger a Chk2 dependent block at the Drosophila midblastula transition. AB - The 13 syncytial cleavage divisions that initiate Drosophila embryogenesis are under maternal genetic control. The switch to zygotic regulation of development at the midblastula transition (MBT) follows mitosis 13, when the cleavage divisions terminate, transcription increases and the blastoderm cellularizes. Embryos mutant for grp, which encodes Checkpoint kinase 1 (Chk1), are DNA replication-checkpoint defective and fail to cellularize, gastrulate or to initiate high-level zygotic transcription at the MBT. The mnk (also known as loki) gene encodes Checkpoint kinase 2 (Chk2), which functions in DNA-damage signal transduction. We show that mnk grp double-mutant embryos are replication checkpoint defective but cellularize, gastrulate and activate high levels of zygotic gene expression. We also show that grp mutant embryos accumulate DNA double-strand breaks and that DNA-damaging agents induce a mnk-dependent block to cellularization and zygotic gene expression. We conclude that the DNA-replication checkpoint maintains genome integrity during the cleavage divisions, and that checkpoint mutations lead to DNA damage that induces a novel Chk2-dependent block at the MBT. PMID- 17409118 TI - The regulation of genes and genomes by small RNAs. AB - A recent Keystone Symposium on 'MicroRNAs and siRNAs: Biological Functions and Mechanisms' was organized by David Bartel and Shiv Grewal (and was held in conjunction with 'RNAi for Target Validation and as a Therapeutic', organized by Stephen Friend and John Maraganore). The 'MicroRNAs and siRNAs' meeting brought together scientists working on diverse biological aspects of small regulatory RNAs, including microRNAs, small interfering RNAs (siRNAs) and Piwi-interacting RNAs (piRNAs and rasiRNAs). Among the themes discussed were the diversity of small regulatory RNAs and their developmental functions, their biogenesis, the identification of their regulatory targets, their mechanisms of action, and their roles in the elaboration of multicellular complexity. PMID- 17409119 TI - The role of NICE technology appraisal in NHS rationing. AB - OBJECTIVE: This article examines the role of National Institute of Health and Clinical Excellence (NICE) technology appraisal in detail, focussing on the process itself and the methods used to establish cost-effective practices for the National Health Service (NHS). AREAS OF AGREEMENT: Approaches to identifying both effective and cost-effective practices have become central to rationing decisions in the NHS. The establishment of the NICE, which produces guidance on what treatments should be provided by the NHS, represents the most visible approach to introducing economic considerations into these decisions. AREAS OF CONTROVERSY: The decisions over which activities will be displaced by NICE approved treatments are made at a local level, while the cost-effectiveness threshold used to evaluate technologies is set nationally. This may result in treatments being displaced which are more cost-effective than those being introduced. AREAS TO DEVELOP RESEARCH: The introduction of programmes looking at disinvestment opportunities to help aid local decision makers is a key step in improving the allocation of NHS resources and removing geographical inequalities. PMID- 17409120 TI - Prophylactic mastectomy: ethical issues. AB - INTRODUCTION: Why in particular should prophylactic mastectomy be seen more as an ethical concern than as a strictly medical question? In this article, four main explanations will be discussed. MAIN POINTS: First, a risky condition is not a disease and prevention does not improve well-being. The benefits are only statistical and make sense at the population level. Secondly, the cause of the risk is a genetic factor and some might argue about genetic 'exceptionalism'. Thirdly, there is no organ as, connected to femininity, sensuality, sexuality, adulthood and motherhood as the breast. Lastly, making tough and complex choices requires assistance from ethics. AREAS OF AGREEMENT: Among ethical principles, western countries often rely on autonomy. The physician has to deliver all the relevant information; based on this 'knowledge' and using their own values, patients will take a decision. AREA OF CONTROVERSY: In 1998 in France, national recommendations set a list of criteria to fulfil, reducing autonomy. EMERGING AREAS FOR DEVELOPING RESEARCH: It might be expected that this tough issue will be solved, thanks to the improvement of prevention and therapeutic efficacy. PMID- 17409121 TI - Experimental acute pancreatitis in PAP/HIP knock-out mice. AB - BACKGROUND AND AIMS: PAP/HIP was first reported as an additional pancreatic secretory protein expressed during the acute phase of pancreatitis. It was shown in vitro to be anti-apoptotic and anti-inflammatory. This study aims to look at whether PAP/HIP plays the same role in vivo. METHODS: A model of caerulein induced pancreatitis was used to compare the outcome of pancreatitis in PAP/HIP( /-) and wild-type mice. RESULTS: PAP/HIP(-/-) mice showed the normal phenotype at birth and normal postnatal development. Caerulein-induced pancreatic necrosis was, however, less severe in PAP/HIP(-/-) mice than in wild-type mice, as judged by lower amylasemia and lipasemia levels and smaller areas of necrosis. On the contrary, pancreas from PAP/HIP(-/-) mice was more sensitive to apoptosis, in agreement with the anti-apoptotic effect of PAP/HIP in vitro. Surprisingly, pancreatic inflammation was more extensive in PAP/HIP(-/-) mice, as judged from histological parameters, increased myeloperoxidase activity and increased pro inflammatory cytokine expression. This result, in apparent contradiction with the limited necrosis observed in these mice, is, however, in agreement with the anti inflammatory function previously reported in vitro for PAP/HIP. This is supported by the observation that activation of the STAT3/SOCS3 pathway was strongly decreased in the pancreas of PAP/HIP(-/-) mice and by the reversion of the apoptotic and inflammatory phenotypes upon administration of recombinant PAP/HIP to PAP/HIP(-/-) mice. CONCLUSION: The anti-apoptotic and anti-inflammatory functions described in vitro for PAP/HIP have physiological relevance in the pancreas in vivo during caerulein-induced pancreatitis. PMID- 17409122 TI - Is surveillance of the small bowel indicated for Lynch syndrome families? AB - BACKGROUND: Small bowel cancer (SBC) is one of the tumours associated with Lynch syndrome (LS). To advise on screening for this tumour it is paramount to be informed about the lifetime risk. The aim of this study was to calculate the lifetime risk of SBC in LS and to identify possible risk factors. METHODS: Clinical and pathological data were collected on 1496 proven or putative carriers of a mismatch repair gene mutation from 189 families. Kaplan-Meier survival analysis was used to calculate the lifetime risk and to assess potential risk factors. RESULTS: 28 (1.9%) of the 1496 (putative) mutation carriers were identified with SBC. The median age at diagnosis was 52 years (range 23-69 years). The lifetime risk of developing SBC was 4.2%. There was no difference in risk between males and females (log rank: p = 0.2470), or between MLH1 and MSH2 mutation carriers (log rank: p = 0.2754). SBC was not observed in MSH6 mutation carriers (n = 203). The previous occurrence of colorectal cancer and a family history of SBC did not increase the risk significantly. CONCLUSIONS: Approximately, one out of 25 mutation carriers will develop SBC during life. No specific risk factors were identified. The risk appeared to be too low to advise screening by means of an invasive burdensome procedure like double balloon enteroscopy. However, screening by a non-invasive procedure (videocapsule endoscopy) might be considered if future studies will show its cost effectiveness. In patients with unexplained abdominal complaints and/or unexplained iron deficiency anaemia SBC should be considered. PMID- 17409123 TI - The deleterious effects of bed rest on human skeletal muscle fibers are exacerbated by hypercortisolemia and ameliorated by dietary supplementation. AB - Prolonged inactivity associated with bed rest in a clinical setting or spaceflight is frequently associated with hypercortisolemia and inadequate caloric intake. Here, we determined the effect of 28 days of bed rest (BR); bed rest plus hypercortisolemia (BRHC); and bed rest plus essential amino acid (AA) and carbohydrate (CHO) supplement (BRAA) on the size and function of single slow- and fast-twitch muscle fibers. Supplementing meals, the BRAA group consumed 16.5 g essential amino acids and 30 g sucrose at 1100, 1600, and 2100 h, and the BRHC subjects received 5 daily doses of 10-15 mg of oral hydrocortisone sodium succinate throughout bed rest. Bed rest induced atrophy and loss of force (mN) and power (muN.FL.s(-1)) in single fibers was exacerbated by hypercortisolemia where soleus peak force declined by 23% in the type I fiber from a prevalue of 0.78 +/- 0.02 to 0.60 +/- 0.02 mN post bed rest (compared to a 7% decline with bed rest alone) and 27% in the type II fiber (1.10 +/- 0.08 vs. 0.81 +/- 0.05 mN). In the BRHC group, peak power dropped by 19, 15, and 11% in the soleus type I, and vastus lateralis (VL) type I and II fibers, respectively. The AA/CHO supplement protected against the bed rest-induced loss of peak force in the type I soleus and peak power in the VL type II fibers. These results provide evidence that an AA/CHO supplement might serve as a successful countermeasure to help preserve muscle function during periods of relative inactivity. PMID- 17409124 TI - Role of the scaffold protein RACK1 in apical expression of CFTR. AB - Previous studies from this laboratory demonstrated a role for protein kinase C (PKC)epsilon in the regulation of cAMP-dependent cystic fibrosis transmembrane regulator (CFTR) Cl channel function via binding of PKCepsilon to RACK1, a receptor for activated C kinase, and of RACK1 to human Na(+)/H(+) exchanger regulatory factor (NHERF1). In the present study, we investigated the role of RACK1 in regulating CFTR function in a Calu-3 airway epithelial cell line. Confocal microscopy and biotinylation of apical surface proteins demonstrate apical localization of RACK1 independent of actin. Mass spectrometric analysis of NHERF1 revealed copurification of tubulin, which, in in vitro binding assays, selectively binds to NHERF1, but not RACK1, via a PDZ1 domain. In binding and pulldown assays, we show direct binding of a PDZ2 domain to NHERF1, pulldown of endogenous NHERF1 by a PDZ2 domain, and inhibition of NHERF1-tubulin binding by a PDZ1 domain. Downregulation of RACK1 using double-stranded silencing RNA reduced the amount of RACK1 by 77.5% and apical expression of biotinylated CFTR by 87.4%. Expression of CFTR, NHERF1, and actin were not altered by treatment with siRACK1 or by nontargeting control silencing RNA, which, in addition, did not affect RACK1 expression. On the basis of these results, we model a RACK1 proteome consisting of PKCepsilon-RACK1-NHERF1-NHERF1-tubulin with a role in stable expression of CFTR in the apical plasma membrane of epithelial cells. PMID- 17409125 TI - Chemokine and chemoattractant receptor expression: post-transcriptional regulation. AB - The magnitude and character of the inflammatory process are determined in part via the trafficking of leukocytes into sites of injury and infection, and this process depends on proper control of the expression of genes encoding chemoattractant peptides and their receptors. Although these controls operate at multiple mechanistic levels, recent evidence indicates that post-transcriptional events governing the half-life of select mRNAs are important determinants. Adenine-uridine rich elements (AREs) located within 3' untranslated regions (UTRs) confer constitutive mRNA instability and in some cases, stabilization following stimulation by ligands of the Toll-IL-1 receptor (TIR) family. Although the importance of AREs in determining activity and mRNA half-life is well recognized, the mechanistic scope and diversity remain poorly understood. Using the mouse KC or CXCL1 gene as a model, we have demonstrated that the abundance of mRNA and protein produced during an inflammatory response depends on multiple mechanistically distinct AREs present in the 3' UTR of the mRNA. The mRNA encoding the receptor for N-terminal formyl-methionine-containing peptides is also unstable and subject to stabilization in response to TIR ligands. These two models can, however, be readily distinguished from one another on the basis of specific stimulus sensitivity and the signaling pathways, through which such stimuli couple to the control of mRNA decay. These models demonstrate the substantial diversity operative in the post-transcriptional regulation of inflammatory gene expression. PMID- 17409126 TI - Use of the QuantiFERON TB Gold test as part of a screening programme in patients with RA under consideration for treatment with anti-TNF-alpha agents: the Newcastle (UK) experience. PMID- 17409127 TI - Corticosteroid-induced lipodystrophy is associated with features of the metabolic syndrome. AB - OBJECTIVE: Familial and HIV-associated lipodystrophies are associated with metabolic disorders and elevated blood pressure. Whether corticosteroid-induced lipodystrophy (CIL) is also associated with features of the metabolic syndrome is unknown. METHODS: We conducted a prospective study in two French tertiary centres and enrolled all consecutive patients starting long-term (> or = 3 months), high dosage (> or = 20 mg/day) systemic corticosteroid therapy. Three investigators assessed the development of CIL from standardized head and neck photographs. Arterial blood pressure and fasting blood glucose levels were assessed at baseline and then every three months until month 12. Total, HDL- and LDL cholesterol and triglycerides were recorded at baseline, month 3 and month 12. RESULTS: Eighty-eight patients were enrolled (women: 75%, mean age: 57.4 +/- 17.9 yrs, mean baseline dosage of prednisolone: 56 +/- 15 mg/day). Sixty-four patients still received corticosteroids at month 12 (mean prednisolone dosage: 11 +/- 4 mg/day). In intention-to-treat analysis, the cumulative incidence rate of CIL at months 3 and 12 was 61 +/- 8% and 69 +/- 9%, respectively. Baseline characteristics were similar in patients who developed CIL and patients who did not develop CIL during follow-up except with regard to baseline body mass index, which was higher in patients who develop CIL (24.3 +/- 3.7 kg/m2 vs 21.4 +/- 3.2 kg/m2, P= 0.02). Blood pressure was significantly higher in CIL+ patients at month 9 (135/78 mmHg vs 127/73 mmHg) and month 12 (141/81 mmHg vs 128/72 mmHg) visits. Moreover, compared with CIL- patients, CIL+ patients had significantly higher plasma concentrations of fasting blood glucose, triglycerides and total cholesterol and lower HDL-cholesterol concentration during follow-up. CONCLUSIONS: CIL is associated with features of the metabolic syndrome and should then not be considered only as an aesthetic challenge. Further studies are required to test the relation between CIL and cardiovascular events. PMID- 17409128 TI - The cost-effectiveness of mycophenolate mofetil as firstline therapy in active lupus nephritis. AB - OBJECTIVES: Systemic lupus erythematosus (SLE) is an autoimmune disorder that can affect any system of the body. Involvement of the kidneys, lupus nephritis (LN), affects up to 50% of SLE patients during the course of their disease, and is characterized by periods of active disease (flares) and remission. For more severe nephritis, an induction course of immunosuppressive therapy is recommended. Options include intravenous cyclophosphamide (IVC) or mycophenolate mofetil (MMF), followed by a maintenance course, typically of azathioprine. The objective of this study is to determine which therapy results in better quality of life (QoL) for patients and which represents best value for money for finite health service resources. METHODS: A patient-level simulation model is developed to estimate the costs and quality-adjusted life-years (QALYs) of a patient treated with IVC or MMF for an induction period of six months. Efficacy, QoL, resource use and cost data are extracted from the literature and standard databases and supplemented with expert opinion where necessary. RESULTS: On average, the model predicts MMF to result in improved QoL compared with IVC. MMF is also less expensive than IVC, costing pound 1600 (euro 2400; US$ 3100) less over the period, based on 2005 NHS prices. The major determinant and cost driver of this result is the requirement for a day-case procedure to administer IVC. Sensitivity analysis shows an 81% probability that MMF will be cost-effective compared with IVC at a willingness to pay of pound 30,000 (euro 44,700; US$ 58,500) per QALY gained. CONCLUSION: MMF is likely to result in better QoL and be less expensive than IVC as induction therapy for LN. PMID- 17409129 TI - Leflunomide versus methotrexate in Wegener's granulomatosis. PMID- 17409130 TI - Resuscitation shoulder--a complication of cardiac arrest. PMID- 17409131 TI - Renal tubular acidosis associated with leflunomide. PMID- 17409132 TI - Mutations of the tissue non-specific alkaline phosphatase gene (TNAP) causing a non-lethal case of perinatal hypophosphatasia. PMID- 17409133 TI - Combined tumour necrosis factor-alpha and tumour necrosis factor receptor genotypes could predict rheumatoid arthritis patients' response to anti-TNF-alpha therapy and explain controversies of studies based on a single polymorphism. PMID- 17409134 TI - Staphylococcal toxic-shock-syndrome-toxin-1 as a risk factor for disease relapse in Wegener's granulomatosis. AB - OBJECTIVES: Nasal carriage of Staphylococcus aureus constitutes a risk factor for disease exacerbation in Wegener's granulomatosis (WG). We hypothesized that staphylococcal superantigens (SAg) are a determinant of S. aureus-related risk for disease relapse in WG. METHODS: In a retrospective longitudinal cohort study in 62 WG patients, we investigated the presence of the staphylococcal SAg genes sea, seb, sec, sed, see, tsst-1 and eta in S. aureus strains isolated from WG patients during an observation period of seven years. Subsequently, we assessed whether relapses of WG were associated with the presence of SAg-positive staphylococci. RESULTS: Of 1718 swab cultures analysed, 709 (41.2%) were S. aureus-positive. Fifty-one patients carried S. aureus, of whom 37 (72.5%) patients carried at least one SAg-positive S. aureus strain. Of the 709 S. aureus positive cultures, 326 (46%) contained at least one SAg gene. Except for see, all assessed SAg genes were detected. sea was found most frequently, followed by sec, tsst-1 and eta and finally, by sed and seb. Using a multivariate, time-dependent Cox regression analysis we found that the presence of S. aureus was associated with relapses of WG (RR 3.2; 95% CI 1.2-8.4). The risk for relapse was modulated by the presence and type of SAg, with tsst-1 being associated with an increased risk for relapse (RR 13.3, 95% CI 4.2-42.6). CONCLUSION: The risk for relapse of WG increases with the presence of tsst-1-positive S. aureus. Eradication of tsst 1-positive S. aureus in WG may show whether disease relapses can be prevented. PMID- 17409135 TI - Ankle/hindfoot arthrodesis in rheumatoid arthritis improves kinematics and kinetics of the knee and hip: a prospective gait analysis study. AB - OBJECTIVES: To evaluate the effects of ankle/hindfoot arthrodesis in rheumatoid arthritis (RA) patients on gait pattern of the knee and hip. METHODS: In this prospective follow-up study, 14 RA patients scheduled for ankle/hindfoot arthrodesis (talo-calcaneal, talo-navicular, calcaneo-cuboid and/or talo-crural joints) and 14 age- and sex-matched healthy controls were included. Three dimensional gait analyses of joint angles, moments and work were performed at the index operation and after 13 months of follow-up. Each patient underwent clinical assessments of pain while walking, overall evaluation of disease activity, Health Related Quality of Life Questionnaire (EQ-5D), activity limitations, maximum walking distance, difficulty with walking surface and gait abnormality. For comparisons of pre- vs post-operative conditions, Wilcoxon's matched pairs test and Friedman ANOVA by rank test were used. RESULTS: At follow-up after ankle/hindfoot fusion surgery, RA patients demonstrated a statistically significant improvement in mean range of joint motions, moments and work in the overlying joints such as the knee and hip. Moreover, there was significantly less pain, disease activity, activity limitation, difficulty with walking surface and gait abnormality. EQ-5D and maximum walking distance were also significantly improved at follow-up. CONCLUSIONS: Our results demonstrate that ankle/hindfoot arthrodesis in RA is an effective intervention to reduce pain and to improve Health Related Quality of Life and functional ability. Moreover, the overlying leg joints experience an improvement in joint motion, muscle-generated joint moments and work during walking. Three-dimensional gait analysis may assist future investigations of the effects of orthopaedic surgery on functional mobility in RA to prevent irreversible disablement. PMID- 17409136 TI - Identification of a novel C-terminal cleavage of Crimean-Congo hemorrhagic fever virus PreGN that leads to generation of an NSM protein. AB - The structural glycoproteins of Crimean-Congo hemorrhagic fever virus (CCHFV; genus Nairovirus, family Bunyaviridae) are derived through endoproteolytic cleavage of a 1,684-amino-acid M RNA segment-encoded polyprotein. This polyprotein is cotranslationally cleaved into the PreGN and PreGC precursors, which are then cleaved by SKI-1 and a SKI-1-like protease to generate the N termini of GN and GC, respectively. However, the resulting polypeptide defined by the N termini of GN and GC is predicted to be larger (58 kDa) than mature GN (37 kDa). By analogy to the topologically similar M segment-encoded polyproteins of viruses in the Orthobunyavirus genus, the C-terminal region of PreGN that contains four predicted transmembrane domains may also contain a nonstructural protein, NSM. To characterize potential PreGN C-terminal cleavage events, a panel of epitope-tagged PreGN truncation and internal deletion mutants was developed. These constructs allowed for the identification of a C-terminal endoproteolytic cleavage within, or very proximal to, the second predicted transmembrane domain following the GN ectodomain and the subsequent generation of a C-terminal fragment. Pulse-chase experiments showed that PreGN C-terminal cleavage occurred shortly after synthesis of the precursor and prior to generation of the GN glycoprotein. The resulting fragment trafficked to the Golgi compartment, the site of virus assembly. Development of an antiserum specific to the second cytoplasmic loop of PreGN allowed detection of cell-associated NSM proteins derived from transient expression of the complete CCHFV M segment and also in the context of virus infection. PMID- 17409137 TI - Lassa virus infection in experimentally infected marmosets: liver pathology and immunophenotypic alterations in target tissues. AB - Lassa virus causes thousands of deaths annually in western Africa and is considered a potential biological weapon. In an attempt to develop a small nonhuman primate model of Lassa fever, common marmosets were subcutaneously inoculated with Lassa virus strain Josiah. This inoculation resulted in a systemic disease with clinical and morphological features mirroring those in fatal human Lassa infection: fever, weight loss, high viremia and viral RNA load in tissues, elevated liver enzymes, and severe morbidity between days 15 and 20. The most prominent histopathology findings included multifocal hepatic necrosis with mild inflammation and hepatocyte proliferation, lymphoid depletion, and interstitial nephritis. Cellular aggregates in regions of hepatocellular necrosis were largely composed of HAM56-positive macrophages, devoid of CD3-positive and CD20-positive cells, and characterized by marked reductions in the intensity of HLA-DP, DQ, DR staining. A marked reduction in the major histocompatibility complex class II expression was also observed in the lymph nodes. Immunophenotypic alterations in spleen included reductions in overall numbers of CD20-positive and CD3-positive cells and the disruption of lymphoid follicular architecture. These findings identify the common marmoset as an appropriate model of human Lassa fever and present the first experimental evidence that replication of Lassa virus in tissues is associated with alterations that would be expected to impair adaptive immunity. PMID- 17409138 TI - Human T-cell leukemia virus type 1 integration target sites in the human genome: comparison with those of other retroviruses. AB - Retroviral integration into the host genome is not entirely random, and integration site preferences vary among different retroviruses. Human immunodeficiency virus (HIV) prefers to integrate within active genes, whereas murine leukemia virus (MLV) prefers to integrate near transcription start sites and CpG islands. On the other hand, integration of avian sarcoma-leukosis virus (ASLV) shows little preference either for genes, transcription start sites, or CpG islands. While host cellular factors play important roles in target site selection, the viral integrase is probably the major viral determinant. It is reasonable to hypothesize that retroviruses with similar integrases have similar preferences for target site selection. Although integration profiles are well defined for members of the lentivirus, spumaretrovirus, alpharetrovirus, and gammaretrovirus genera, no members of the deltaretroviruses, for example, human T cell leukemia virus type 1 (HTLV-1), have been evaluated. We have mapped 541 HTLV 1 integration sites in human HeLa cells and show that HTLV-1, like ASLV, does not specifically target transcription units and transcription start sites. Comparing the integration sites of HTLV-1 with those of ASLV, HIV, simian immunodeficiency virus, MLV, and foamy virus, we show that global and local integration site preferences correlate with the sequence/structure of virus-encoded integrases, supporting the idea that integrase is the major determinant of retroviral integration site selection. Our results suggest that the global integration profiles of other retroviruses could be predicted from phylogenetic comparisons of the integrase proteins. Our results show that retroviruses that engender different insertional mutagenesis risks can have similar integration profiles. PMID- 17409139 TI - MEK1/2 inhibitors block basal and transforming growth factor 1beta1-stimulated JC virus multiplication. AB - The multiplication of the human neurotropic polyomavirus JC virus (JCV) is regulated by cell membrane receptors and nuclear transcription factors. Signaling pathways also play a role in determining the extent to which JCV can productively infect cells. These data show that constitutively active MEK1 protein (CA-MEK1), overexpressed in cultures of human glia, supports a substantial increase in late JCV protein (Vp-1) synthesis. The specificity of this pathway was indicated by no significant enhancement of JCV multiplication through activation of other components of mitogen-activated protein kinase pathways such as p38, Jun N terminal protein kinase, and protein kinase A. Further evidence supporting the importance of signaling in JCV infection came from addition of transforming growth factor beta1 (TGF-beta1), which stimulated a 200% increase of Vp-1 expression. Specific MEK1/2 inhibitors, flavenoid PD98059 and U0126, decreased the basal and TGF-beta1-stimulated Vp-1 expression by 95% or more. TGF-beta1 is known to phosphorylate/activate Smad DNA binding proteins that could subsequently bind or increase binding to JCV promoter sequences, linking the effects of signaling with JCV transcriptional regulation. The effectiveness with which MEK1/2 inhibitors block JCV multiplication provides insight that may contribute to development of compounds directed against JCV. PMID- 17409140 TI - Direct comparison of antigen production and induction of apoptosis by canarypox virus- and modified vaccinia virus ankara-human immunodeficiency virus vaccine vectors. AB - Recombinant poxvirus vectors are undergoing intensive evaluation as vaccine candidates for a variety of infectious pathogens. Avipoxviruses, such as canarypox virus, are replication deficient in mammalian cells by virtue of a poorly understood species-specific restriction. Highly attenuated vaccinia virus strains such as modified vaccinia virus Ankara (MVA) are similarly unable to complete replication in most mammalian cells but have an abortive-late phenotype, in that the block to replication occurs post-virus-specific DNA replication. In this study, an identical expression cassette for human immunodeficiency virus gag, pro, and env coding sequences was placed in canarypox virus and MVA vector backbones in order to directly compare vector-borne expression and to analyze differences in vector-host cell interactions. Antigen production by recombinant MVA was shown to be greater than that from recombinant canarypox virus in the mammalian cell lines and in the primary human cells tested. This observation was primarily due to a longer duration of antigen production in recombinant MVA infected cells. Apoptosis induction was found to be more profound with the empty canarypox virus vector than with MVA. Remarkably, however, the inclusion of a gag/pro/env expression cassette altered the kinetics of apoptosis induction in recombinant MVA-infected cells to levels equal to those found in canarypox virus infected cells. Antigen production by MVA was noted to be greater in human dendritic cells and resulted in enhanced T-cell stimulation in an in vitro antigen presentation assay. These results reveal differences in poxvirus vector host cell interactions that should be relevant to their use as immunization vehicles. PMID- 17409141 TI - Base pairing between cis-acting sequences contributes to template switching during plus-strand DNA synthesis in human hepatitis B virus. AB - Hepadnaviruses utilize two template switches (primer translocation and circularization) during synthesis of plus-strand DNA to generate a relaxed circular (RC) DNA genome. In duck hepatitis B virus (DHBV) three cis-acting sequences, 3E, M, and 5E, contribute to both template switches through base pairing, 3E with the 3' portion of M and 5E with the 5' portion of M. Human hepatitis B virus (HBV) also contains multiple cis-acting sequences that contribute to the accumulation of RC DNA, but the mechanisms through which these sequences contribute were previously unknown. Three of the HBV cis-acting sequences (h3E, hM, and h5E) occupy positions equivalent to those of the DHBV 3E, M, and 5E. We present evidence that h3E and hM contribute to the synthesis of RC DNA through base pairing during both primer translocation and circularization. Mutations that disrupt predicted base pairing inhibit both template switches while mutations that restore the predicted base pairing restore function. Therefore, the h3E-hM base pairing appears to be a conserved requirement for template switching during plus-strand DNA synthesis of HBV and DHBV. Also, we show that base pairing is not sufficient to explain the mechanism of h3E and hM, as mutating sequences adjacent to the base pairing regions inhibited both template switches. Finally, we did not identify predicted base pairing between h5E and the hM region, indicating a possible difference between HBV and DHBV. The significance of these similarities and differences between HBV and DHBV will be discussed. PMID- 17409142 TI - Characterization of protein-protein interactions critical for poliovirus replication: analysis of 3AB and VPg binding to the RNA-dependent RNA polymerase. AB - Two critical interactions within the poliovirus RNA replication complex are those of the RNA-dependent RNA polymerase 3D with the viral proteins 3AB and VPg. 3AB is a membrane-binding protein responsible for the localization of the polymerase to the membranous vesicles at which replication occurs. VPg (a peptide comprising the 3B region of 3AB) is the 22-residue soluble product of 3AB cleavage and serves as the protein primer for RNA replication. The detailed interactions of these proteins with the RNA-dependent RNA polymerase 3D were analyzed to elucidate the precise roles of 3AB and VPg in the viral RNA replication complex. Using a membrane-based pull-down assay, we have identified a binding "hot-spot" spanning residues 100 to 104 in the 3B (VPg) region of 3AB which plays a critical role in mediating the interaction of 3AB with the polymerase. Isothermal titration calorimetry shows that the interaction of VPg with 3D is enthalpically driven, with a dissociation constant of 11 microM. Mutational analyses of VPg indicate that a subset of the residues important for 3AB-3D binding are also important for VPg-3D binding. Two residues in particular, P14 and R17, were shown to be absolutely critical for the binding interaction. This work provides the direct characterization of two binding interactions critical for the replication of this important class of viruses and identifies a conserved polymerase binding sequence responsible for targeting the polymerase. PMID- 17409143 TI - Association of vaccinia virus fusion regulatory proteins with the multicomponent entry/fusion complex. AB - The proteins encoded by the A56R and K2L genes of vaccinia virus form a heterodimer (A56/K2) and have a fusion regulatory role as deletion or mutation of either causes infected cells to form large syncytia spontaneously. Here, we showed that syncytia formation is dependent on proteins of the recently described entry fusion complex (EFC), which are also required for virus-cell fusion and low pH-triggered cell-cell fusion. This finding led us to consider that A56/K2 might prevent fusion by direct or indirect interaction with the EFC. To test this hypothesis, we made a panel of recombinant vaccinia viruses that have a tandem affinity purification tag attached to A56, K2, or the A28 EFC protein. Interaction between A56/K2 and the EFC was demonstrated by their copurification from detergent-treated lysates of infected cells and identification by mass spectrometry or Western blotting. In addition, a purified soluble transmembrane deleted form of A56/K2 was shown to interact with the EFC. Tagged A56 did not interact with the EFC in the absence of K2, nor did tagged K2 interact with the EFC in the absence of A56. The finding that both A56 and K2 are required for efficient binding to the EFC fits well with prior experiments showing that mutation of either A56 or K2 results in spontaneous fusion of infected cells. Because A56 and K2 are located on the surface of infected cells, they are in position to interact with the EFC of released progeny virions and prevent back fusion and syncytia formation. PMID- 17409144 TI - The ATM/ATR signaling effector Chk2 is targeted by Epstein-Barr virus nuclear antigen 3C to release the G2/M cell cycle block. AB - Epstein-Barr virus (EBV) infects most of the human population and persists in B lymphocytes for the lifetime of the host. The establishment of latent infection by EBV requires the expression of a unique repertoire of genes. The product of one of these viral genes, the EBV nuclear antigen 3C (EBNA3C), is essential for the growth transformation of primary B lymphocytes in vitro and can regulate the transcription of a number of viral and cellular genes important for the immortalization process. This study demonstrates an associated function of EBNA3C which involves the disruption of the G2/M cell cycle checkpoint. We show that EBNA3C-expressing lymphoblastoid cell lines treated with the drug nocodazole, which is known to block cells at the G2/M transition, did not show a G2/M specific checkpoint arrest. Analyses of the cell cycles of cells expressing EBNA3C demonstrated that the expression of this essential EBV nuclear antigen is capable of releasing the G2/M checkpoint arrest induced by nocodazole. This G2/M arrest in response to nocodazole was also abolished by caffeine, suggesting an involvement of the ATM/ATR signaling pathway in the regulation of this cell cycle checkpoint. Importantly, we show that the direct interaction of EBNA3C with Chk2, the ATM/ATR signaling effector, is responsible for the release of this nocodazole induced G2/M arrest and that this interaction leads to the serine 216 phosphorylation of Cdc25c, which is sequestered in the cytoplasm by 14-3-3. Overall, our data suggest that EBNA3C can directly regulate the G2/M component of the host cell cycle machinery, allowing for the release of the checkpoint block. PMID- 17409145 TI - In vivo study of the HC-TN strain of hepatitis C virus recovered from a patient with fulminant hepatitis: RNA transcripts of a molecular clone (pHC-TN) are infectious in chimpanzees but not in Huh7.5 cells. AB - Both viral and host factors are thought to influence the pathogenesis of hepatitis C virus (HCV) infection. We studied strain HC-TN (genotype 1a), which caused fulminant hepatic failure in a patient and, subsequently, severe hepatitis in a chimpanzee (CH1422), to analyze the relationship between disease severity, host immune response, viral evolution, and outcome. A second chimpanzee (CH1581) was infected from CH1422 plasma, and a third chimpanzee (CH1579) was infected from RNA transcripts of a consensus cDNA of HC-TN (pHC-TN). RNA transcripts of pHC-TN did not replicate in Huh7.5 cells, which were recently found to be susceptible to infection with another fulminant HCV strain (JFH1). The courses of viremia were similar in the three animals. However, CH1581 and CH1579 developed a less severe acute hepatitis than CH1422. CH1579 and CH1422 resolved the infection, whereas CH1581 became persistently infected. CH1579 and CH1581, despite their differing outcomes, both developed significant intrahepatic cellular immune responses, but not antibodies to the envelope glycoproteins or neutralizing antibodies, during the acute infection. We analyzed the polyprotein sequences of virus recovered at five and nine time points from CH1579 and CH1581, respectively, during the first year of follow-up. High mutation rates and high proportions of nonsynonymous mutations suggested immune pressure and positive selection in both animals. Changes were not selected until after the initial decrease in virus titers and after the development of immune responses and hepatitis. Subsequently, however, mutations emerged repeatedly in both animals. Overall, our results indicate that disease severity and outcome of acute HCV infection depend primarily on the host response. PMID- 17409146 TI - Crystal structures of major envelope proteins VP26 and VP28 from white spot syndrome virus shed light on their evolutionary relationship. AB - White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 A, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelope proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt beta-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core beta barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV. PMID- 17409147 TI - Therapy with minocycline aggravates experimental rabies in mice. AB - Minocycline is a tetracycline derivative with antiapoptotic and anti-inflammatory properties, and the drug has been shown to have beneficial effects in a variety of models of neurological disorders. The potentially neuroprotective role of minocycline was assessed in experimental in vitro and in vivo models of rabies virus infection. In this study, 5 nM minocycline did not improve the viability of embryonic mouse cortical and hippocampal neurons infected in vitro with the attenuated SAD-D29 strain of rabies virus, based on assessments using trypan blue exclusion. Two-day-old ICR mice were inoculated in the right hind limb thigh muscle with SAD-D29, and they received daily subcutaneous injections of either 50 mg/kg minocycline or vehicle (phosphate-buffered saline). Infected minocycline treated mice experienced an earlier onset of neurologic signs and greater mortality (83% versus 50%) than those receiving vehicle (log rank test, P=0.002 and P=0.003, respectively). Immunohistochemical analysis of rabies virus antigen distribution was performed at early time points and in moribund mice. There were greater numbers of infected neurons in the regional brain areas of minocycline treated mice than in vehicle-treated mice, which was significant in the CA1 region of the hippocampus. There was less apoptosis (P=0.01) and caspase 3 immunostaining (P=0.0008) in the midbrains of mice treated with minocycline than in mice treated with vehicle, consistent with a neuroprotective role of neuronal apoptosis that may have had a mild effect of inhibiting viral spread. Reduced infiltration of CD3+ T cells was observed in the pons/medulla of moribund mice that received minocycline therapy (P=0.008), suggesting that the anti inflammatory actions of minocycline may intensify the neurologic disease. These findings indicate that minocycline has important detrimental effects in the therapy of experimental rabies. Empirical therapy with minocycline should therefore be approached with caution in cases of human rabies and possibly other viral encephalitides until more experimental data become available. PMID- 17409148 TI - Bcl-2 blocks accretion or depletion of stored calcium but has no effect on the redistribution of IP3 receptor I mediated by glycoprotein E of herpes simplex virus 1. AB - We examined the status of stable, resting intracellular Ca2+ ([Ca2+]i) and the calcium that can be released from intracellular stores in HEp-2 or VAX-3 cells overexpressing Bcl-2 after infection with wild-type or mutant herpes simplex viruses. The mutants included viruses lacking ICP4 or ICP27 and known to induce apoptosis. We report the following. Stable Ca2+ levels decrease after infection with wild-type or mutant viruses in both HEp-2 and VAX-3 cells. The histamine sensitive calcium stores became depleted in wild-type and mutant virus-infected cells late in infection but increased significantly in DeltaICP4- or DeltaICP27 infected cells prior to depletion. In VAX-3 cells, the depletion in calcium stores did not take place as late as 24 h after infection, concomitant with lack of visually detectable cytopathic effects. Concurrent analyses showed that the amounts of IP3 Ca2+-receptor type I (IP3R-I) remained stable throughout infection, but the intensity of the signal increased and intracellular distribution changed dramatically in both HEp-2 and VAX-3 cells infected with the wild-type and all mutant viruses, except for the mutant lacking glycoprotein E (DeltagE). In transfected HEp-2 cells, gE and gI were more effective at augmenting the signal intensity and redistribution of IP3R-I than gE or gI alone. We conclude the following. (i) Depleted histamine-sensitive calcium stores correlate with appearance of cytopathic effects. (ii) Apoptosis, the calcium stores, and cytopathic effects are regulated by Bcl-2. (iii) The changes in the distribution of IP3R-I are mediated by the viral Fc receptor complex, but the redistribution is not related to changes in stored calcium. PMID- 17409149 TI - The cytomegalovirus-specific CD4+ T-cell response expands with age and markedly alters the CD4+ T-cell repertoire. AB - Immune function in the elderly is associated with a number of phenotypic and functional abnormalities, and this phenomenon of immune senescence is associated with increased susceptibility to infection. The immune response to pathogens frequently declines with age, but the CD8(+) T-cell response to cytomegalovirus (CMV) is unusual, as it demonstrates a significant expansion over time. Here we have documented the CD4(+) T-cell immune response to CMV in healthy donors of different ages. The magnitude of the CMV-specific CD4(+) T-cell immune response increases from a mean of 2.2% of the CD4(+) T-cell pool in donors below 50 years of age to 4.7% in donors aged over 65 years. In addition, CMV-specific CD4(+) T cells in elderly donors demonstrate decreased production of interleukin-2 and less dependence on costimulation. CMV seropositivity is associated with marked changes in the phenotype of the overall CD4(+) T-cell repertoire in healthy aged donors, including an increase in CD57(+) expression and a decrease in CD28 and CD27 expression, a phenotypic profile characteristic of immune senescence. This memory inflation of CMV-specific CD4(+) T cells contributes to evidence that CMV infection may be damaging to immune function in elderly individuals. PMID- 17409150 TI - Crystal structure of a monomeric form of severe acute respiratory syndrome coronavirus endonuclease nsp15 suggests a role for hexamerization as an allosteric switch. AB - Mature nonstructural protein-15 (nsp15) from the severe acute respiratory syndrome coronavirus (SARS-CoV) contains a novel uridylate-specific Mn2+ dependent endoribonuclease (NendoU). Structure studies of the full-length form of the obligate hexameric enzyme from two CoVs, SARS-CoV and murine hepatitis virus, and its monomeric homologue, XendoU from Xenopus laevis, combined with mutagenesis studies have implicated several residues in enzymatic activity and the N-terminal domain as the major determinant of hexamerization. However, the tight link between hexamerization and enzyme activity in NendoUs has remained an enigma. Here, we report the structure of a trimmed, monomeric form of SARS-CoV nsp15 (residues 28 to 335) determined to a resolution of 2.9 A. The catalytic loop (residues 234 to 249) with its two reactive histidines (His 234 and His 249) is dramatically flipped by approximately 120 degrees into the active site cleft. Furthermore, the catalytic nucleophile Lys 289 points in a diametrically opposite direction, a consequence of an outward displacement of the supporting loop (residues 276 to 295). In the full-length hexameric forms, these two loops are packed against each other and are stabilized by intimate intersubunit interactions. Our results support the hypothesis that absence of an adjacent monomer due to deletion of the hexamerization domain is the most likely cause for disruption of the active site, offering a structural basis for why only the hexameric form of this enzyme is active. PMID- 17409151 TI - The Kaposi's sarcoma-associated herpesvirus K5 E3 ubiquitin ligase modulates targets by multiple molecular mechanisms. AB - Kaposi's sarcoma-associated herpesvirus encodes two highly related membrane associated, RING-CH-containing (MARCH) family E3 ubiquitin ligases, K3 and K5, that can down regulate a variety of cell surface proteins through enhancement of their endocytosis and degradation. In this report we present data that while K5 modulation of major histocompatibility complex class I (MHC-I) closely mirrors the mechanisms used by K3, alternative molecular pathways are utilized by this E3 ligase in the down regulation of intercellular adhesion molecule 1 (ICAM-1) and B7.2. Internalization assays demonstrate that down regulation of each target can occur through increased endocytosis from the cell surface. However, mutation of a conserved tyrosine-based endocytosis motif in K5 resulted in a protein lacking the ability to direct an increased rate of MHC-I or ICAM-1 internalization but still able to down regulate B7.2 in a ubiquitin-dependent but endocytosis independent manner. Further, mutation of two acidic clusters abolished K5 mediated MHC-I degradation while only slightly decreasing ICAM-1 or B7.2 protein destruction. This same mutant abolished detectable ubiquitylation of all targets. These data indicate that while K5 can act as an E3 ubiquitin ligase to directly mediate cell surface molecule destruction, regulation of its targets occurs through multiple pathways, including ubiquitin-independent mechanisms. PMID- 17409152 TI - Indirect regulation of CD4 T-cell responses by tumor necrosis factor receptors in an acute viral infection. AB - Despite the well-recognized importance of CD4 T-cell help in the induction of antibody production and cytotoxic-T-lymphocyte responses, the regulation of CD4 T cell responses is not well understood. Using mice deficient for TNF receptor I (TNFR I) and/or TNFR II, we show that TNFR I and TNFR II play redundant roles in down regulating the expansion of CD4 T cells during an acute infection of mice with lymphocytic choriomeningitis virus (LCMV). Adoptive transfer experiments using T-cell-receptor transgenic CD4 T cells and studies with mixed bone marrow chimeras indicated that indirect effects and not direct effects on T cells mediated the suppressive function of TNF on CD4 T-cell expansion during the primary response. Further studies to characterize the indirect effects of TNF suggested a role for TNFRs in LCMV-induced deletion of CD11c(hi) dendritic cells in the spleen, which might be a mechanism to limit the duration of antigenic stimulation and CD4 T-cell expansion. Consequent to enhanced primary expansion, there was a substantial increase in the number of LCMV-specific memory CD4 T cells in the spleens of mice deficient for both TNFR I and TNFR II. In summary, our findings suggest that TNFRs down regulate CD4 T-cell responses during an acute LCMV infection by a non-T-cell autonomous mechanism. PMID- 17409153 TI - Formation of hepatitis B virus covalently closed circular DNA: removal of genome linked protein. AB - Hepatitis B virus (HBV) contains a small, partially double-stranded, relaxed circular (RC) DNA genome. RC DNA needs to be converted to covalently closed circular (CCC) DNA, which serves as the template for all viral RNA transcription. As a first step toward understanding how CCC DNA is formed, we analyzed the viral and host factors that may be involved in CCC DNA formation, using transient and stable DNA transfections of HBV and the related avian hepadnavirus, duck hepatitis B virus (DHBV). Our results show that HBV CCC DNA formed in hepatoma cells was derived predominantly from RC DNA with a precise junction sequence. In contrast to that of DHBV, HBV CCC DNA formation in cultured cells was accompanied by the accumulation of a RC DNA species from which the covalently attached viral reverse transcriptase (RT) protein was removed (protein-free or PF-RC DNA). Furthermore, whereas envelope deficiency led to increased CCC DNA formation in DHBV, it resulted mainly in increased PF-RC, but not CCC, DNA in HBV, suggesting that the envelope protein(s) may negatively regulate a step in CCC DNA formation that precedes deproteination in both HBV and DHBV. Interestingly, PF-RC DNA, in contrast to RT-linked RC DNA, contained, almost exclusively, mature plus-strand DNA, suggesting that the RT protein was removed preferentially from mature RC DNA. PMID- 17409154 TI - The late promoter of the human cytomegalovirus viral DNA polymerase processivity factor has an impact on delayed early and late viral gene products but not on viral DNA synthesis. AB - Transcription of the DNA polymerase processivity factor gene (UL44) of human cytomegalovirus initiates at three distinct start sites, which are differentially regulated during productive infection. Two of these start sites, the distal and proximal sites, are active at early times, and the middle start site is active at only late times after infection (F. Leach and E. S. Mocarski, J. Virol. 63:1783 1791, 1989). Compared to the wild type, UL44 gene expression was lower for recombinant viruses with the distal or the middle TATA element mutated. The transcripts initiating from the distal or middle start site facilitated late viral gene expression. The level of viral DNA synthesis was affected by mutation of the distal TATA element. In contrast, mutation of the middle TATA element did not affect the level of viral DNA synthesis, but it did affect significantly the level of late viral gene expression. Recombinant viruses with the distal or middle TATA element mutated grew more slowly than the wild type at both low and high multiplicities of infection. Reduced expression of the UL44 gene from the late middle viral promoter correlated with decreased late viral protein expression and decreased viral growth. PMID- 17409155 TI - Productive varicella-zoster virus infection of cultured intact human ganglia. AB - Varicella-zoster virus (VZV) is a species-specific herpesvirus which infects sensory ganglia. We have developed a model of infection of human intact explant dorsal root ganglia (DRG). Following exposure of DRG to VZV, viral antigens were detected in neurons and nonneuronal cells. Enveloped virions were visualized by transmission electron microscopy in neurons and nonneuronal cells and within the extracellular space. Moreover, rather than remaining highly cell associated during infection of cultured cells, such as fibroblasts, cell-free VZV was released from infected DRG. This model enables VZV infection of ganglionic cells to be studied in the context of intact DRG. PMID- 17409157 TI - Effective T-cell responses select human immunodeficiency virus mutants and slow disease progression. AB - The possession of some HLA class I molecules is associated with delayed progression to AIDS. The mechanism behind this beneficial effect is unclear. We tested the idea that cytotoxic T-cell responses restricted by advantageous HLA class I molecules impose stronger selection pressures than those restricted by other HLA class I alleles. As a measure of the selection pressure imposed by HLA class I alleles, we determined the extent of HLA class I-associated epitope variation in a cohort of European human immunodeficiency virus (HIV)-positive individuals (n=84). We validated our findings in a second, distinct cohort of African patients (n=516). We found that key HIV epitopes restricted by advantageous HLA molecules (B27, B57, and B51 in European patients and B5703, B5801, and B8101 in African patients) were more frequently mutated in individuals bearing the restricting HLA than in those who lacked the restricting HLA class I molecule. HLA alleles associated with clinical benefit restricted certain epitopes for which the consensus peptides were frequently recognized by the immune response despite the circulating virus's being highly polymorphic. We found a significant inverse correlation between the HLA-associated hazard of disease progression and the mean HLA-associated prevalence of mutations within epitopes (P=0.028; R2=0.34). We conclude that beneficial HLA class I alleles impose strong selection at key epitopes. This is revealed by the frequent association between effective T-cell responses and circulating viral escape mutants and the rarity of these variants in patients who lack these favorable HLA class I molecules, suggesting a significant pressure to revert. PMID- 17409158 TI - Biological and immunological relations among human parvovirus B19 genotypes 1 to 3. AB - The human parvovirus B19 is now divided into three genotypes: type 1 (prototype), type 2 (A6- and LaLi-like), and type 3 (V9-like). In overall DNA sequence, the three virus types differ by approximately 10%. The most striking DNA dissimilarity, of >20%, is observed within the p6 promoter region. Because of the scarcity of data on the biological activities and pathogenetic potentials of virus types 2 and 3, we examined the functional characteristics of these virus types. We found the activities of the three p6 promoters to be of equal strength and to be most active in B19-permissive cells. Virus type 2 capsid protein VP2, alone or together with VP1, was expressed with the baculovirus system and was shown to assemble into icosahedral parvovirus-like particles, which were reactive in the hemagglutination assay. Furthermore, sera containing DNA of any of the three B19 types were shown to hemagglutinate. The infectivities of these sera were examined in two B19-permissive cell lines. Reverse transcription-PCR revealed synthesis of spliced B19 mRNAs, and immunofluorescence verified the production of NS and VP proteins in the infected cells. All three genotypes showed similar functional characteristics in all experiments performed, showing that the three virus types indeed belong to the same species, i.e., human parvovirus B19. Additionally, the antibody activity in sera from B19 type 1- or type 2-infected subjects (long-term immunity) was examined with homo- and heterologous virus-like particles. Cross-reactivity of 100% was observed, indicating that the two B19 genotypes comprise a single serotype. PMID- 17409159 TI - The transcriptional repressor K-RBP modulates RTA-mediated transactivation and lytic replication of Kaposi's sarcoma-associated herpesvirus. AB - The replication and transcription activator (RTA) protein of Kaposi's sarcoma (KS)-associated herpesvirus (KSHV)/human herpesvirus 8 functions as the key regulator to induce KSHV lytic replication from latency through activation of the lytic cascade of KSHV. Elucidation of the host factors involved in RTA-mediated transcriptional activation is pivotal for understanding the transition between viral latency and lytic replication. KSHV-RTA binding protein (K-RBP) was previously isolated as a cellular RTA binding protein of unknown function. Sequence analysis showed that K-RBP contains a Kruppel-associated box (KRAB) at the N terminus and 12 adjacent zinc finger motifs. In similarity to other KRAB containing zinc finger proteins, K-RBP is a transcriptional repressor. Mutational analysis revealed that the KRAB domain is responsible for the transcriptional suppression activity of this protein and that the repression is histone deacetylase independent. K-RBP was found to repress RTA-mediated transactivation and interact with TIF1beta (transcription intermediary factor 1beta), a common corepressor of KRAB-containing protein, to synergize with K-RBP in repression. Overexpression and knockdown experiment results suggest that K-RBP is a suppressor of RTA-mediated KSHV reactivation. Our findings suggest that the KRAB containing zinc finger protein K-RBP can suppress RTA-mediated transactivation and KSHV lytic replication and that KSHV utilizes this protein as a regulator to maintain a balance between latency and lytic replication. PMID- 17409160 TI - Dissecting the neutralizing antibody specificities of broadly neutralizing sera from human immunodeficiency virus type 1-infected donors. AB - Attempts to elicit broadly neutralizing antibody responses by human immunodeficiency virus type 1 (HIV-1) vaccine antigens have been met with limited success. To better understand the requirements for cross-neutralization of HIV-1, we have characterized the neutralizing antibody specificities present in the sera of three asymptomatic individuals exhibiting broad neutralization. Two individuals were infected with clade B viruses and the third with a clade A virus. The broadly neutralizing activity could be exclusively assigned to the protein A-reactive immunoglobulin G (IgG) fraction of all three donor sera. Neutralization inhibition assays performed with a panel of linear peptides corresponding to the third hypervariable (V3) loop of gp120 failed to inhibit serum neutralization of a panel of HIV-1 viruses. The sera also failed to neutralize chimeric simian immunodeficiency virus (SIV) and HIV-2 viruses displaying highly conserved gp41-neutralizing epitopes, suggesting that antibodies directed against these epitopes likely do not account for the broad neutralizing activity observed. Polyclonal IgG was fractionated on recombinant monomeric clade B gp120, and the neutralization capacities of the gp120-depleted samples were compared to that of the original polyclonal IgG. We found that the gp120-binding antibody population mediated neutralization of some isolates, but not all. Overall, the data suggest that broad neutralization results from more than one specificity in the sera but that the number of these specificities is likely small. The most likely epitope recognized by the monomeric gp120 binding neutralizing fraction is the CD4 binding site, although other epitopes, such as the glycan shield, cannot be excluded. PMID- 17409161 TI - Herpes simplex virus latency-associated transcript sequence downstream of the promoter influences type-specific reactivation and viral neurotropism. AB - Herpes simplex virus (HSV) establishes latency in sensory nerve ganglia during acute infection and may later periodically reactivate to cause recurrent disease. HSV type 1 (HSV-1) reactivates more efficiently than HSV-2 from trigeminal ganglia while HSV-2 reactivates more efficiently than HSV-1 from lumbosacral dorsal root ganglia (DRG) to cause recurrent orofacial and genital herpes, respectively. In a previous study, a chimeric HSV-2 that expressed the latency associated transcript (LAT) from HSV-1 reactivated similarly to wild-type HSV-1, suggesting that the LAT influences the type-specific reactivation phenotype of HSV-2. To further define the LAT region essential for type-specific reactivation, we constructed additional chimeric HSV-2 viruses by replacing the HSV-2 LAT promoter (HSV2-LAT-P1) or 2.5 kb of the HSV-2 LAT sequence (HSV2-LAT-S1) with the corresponding regions from HSV-1. HSV2-LAT-S1 was impaired for reactivation in the guinea pig genital model, while its rescuant and HSV2-LAT-P1 reactivated with a wild-type HSV-2 phenotype. Moreover, recurrences of HSV-2-LAT-S1 were frequently fatal, in contrast to the relatively mild recurrences of the other viruses. During recurrences, HSV2-LAT-S1 DNA increased more in the sacral cord compared to its rescuant or HSV-2. Thus, the LAT sequence region, not the LAT promoter region, provides essential elements for type-specific reactivation of HSV-2 and also plays a role in viral neurotropism. HSV-1 DNA, as quantified by real-time PCR, was more abundant in the lumbar spinal cord, while HSV-2 DNA was more abundant in the sacral spinal cord, which may provide insights into the mechanism for type-specific reactivation and different patterns of central nervous system infection of HSV-1 and HSV-2. PMID- 17409162 TI - Chromatin profiling of Epstein-Barr virus latency control region. AB - Epstein-Barr virus (EBV) escapes host immunity by the reversible and epigenetic silencing of immunogenic viral genes. We previously presented evidence that a dynamic chromatin domain, which we have referred to as the latency control region (LCR), contributes to the reversible repression of EBNA2 and LMP1 gene transcription. We now explore the protein-DNA interaction profiles for a few known regulatory factors and histone modifications that regulate LCR structure and activity. A chromatin immunoprecipitation assay combined with real-time PCR analysis was used to analyze protein-DNA interactions at approximately 500-bp intervals across the first 60,000 bp of the EBV genome. We compared the binding patterns of EBNA1 with those of the origin recognition complex protein ORC2, the chromatin boundary factor CTCF, the linker histone H1, and several histone modifications. We analyzed three EBV-positive cell lines (MutuI, Raji, and LCL3459) with distinct transcription patterns reflecting different latency types. Our findings suggest that histone modification patterns within the LCR are complex but reflect differences in each latency type. The most striking finding was the identification of CTCF sites immediately upstream of the Qp, Cp, and EBER transcription initiation regions in all three cell types. In transient assays, CTCF facilitated EBNA1-dependent transcription activation of Cp, suggesting that CTCF coordinates interactions between different chromatin domains. We also found that histone H3 methyl K4 clustered with CTCF and EBNA1 at sites of active transcription or DNA replication initiation. Our findings support a model where CTCF delineates multiple domains within the LCR and regulates interactions between these domains that correlate with changes in gene expression. PMID- 17409163 TI - Location and role of free cysteinyl residues in the Sindbis virus E1 and E2 glycoproteins. AB - Sindbis virus is a single-stranded positive-sense RNA virus. It is composed of 240 copies of three structural proteins: E1, E2, and capsid. These proteins form a mature virus particle composed of two nested T=4 icosahedral shells. A complex network of disulfide bonds in the E1 and E2 glycoproteins is developed through a series of structural intermediates as virus maturation occurs (M. Mulvey and D. T. Brown, J. Virol. 68:805-812, 1994; M. Carleton et al., J. Virol. 71:1558-1566, 1997). To better understand the nature of this disulfide network, E1 and E2 cysteinyl residues were labeled with iodoacetamide in the native virus particle and analyzed by liquid chromatography-tandem mass spectrometry. This analysis identified cysteinyl residues of E1 and E2, which were found to be label accessible in the native virus particle, as well as those that were either label inaccessible or blocked by their involvement in disulfide bonds. Native virus particles alkylated with iodoacetamide demonstrated a 4-log decrease in viral infectivity. This suggests that the modification of free cysteinyl residues results in the loss of infectivity by destabilizing the virus particle or that a rearrangement of disulfide bonds, which is required for infectivity, is blocked by the modification. Although modification of these residues prevented infectivity, it did not alter the ability of virus to fuse cells after exposure to acidic pH; thus, modification of free cysteinyl residues biochemically separated the process of infection from the process of membrane fusion. PMID- 17409164 TI - Neutralizing antibody responses in acute human immunodeficiency virus type 1 subtype C infection. AB - The study of the evolution and specificities of neutralizing antibodies during the course of human immunodeficiency virus type 1 (HIV-1) infection may be important in the discovery of possible targets for vaccine design. In this study, we assessed the autologous and heterologous neutralization responses of 14 HIV-1 subtype C-infected individuals, using envelope clones obtained within the first 2 months postinfection. Our data show that potent but relatively strain-specific neutralizing antibodies develop within 3 to 12 months of HIV-1 infection. The magnitude of this response was associated with shorter V1-to-V5 envelope lengths and fewer glycosylation sites, particularly in the V1-V2 region. Anti-MPER antibodies were detected in 4 of 14 individuals within a year of infection, while antibodies to CD4-induced (CD4i) epitopes developed to high titers in 12 participants, in most cases before the development of autologous neutralizing antibodies. However, neither anti-MPER nor anti-CD4i antibody specificity conferred neutralization breadth. These data provide insights into the kinetics, potency, breadth, and epitope specificity of neutralizing antibody responses in acute HIV-1 subtype C infection. PMID- 17409165 TI - Activation of pedunculopontine tegmental PKA prevents GABAB receptor activation mediated rapid eye movement sleep suppression in the freely moving rat. AB - The pedunculopontine tegmental (PPT) GABAergic system plays a crucial role in the regulation of rapid eye movement (REM) sleep. I recently reported that the activation of PPT GABA(B) receptors suppressed REM sleep by inhibiting REM-on cells. One of the important mechanisms for GABA(B) receptor activation-mediated physiological action is the inhibition of the intracellular cAMP-dependent protein kinase A (cAMP-PKA) signaling pathway. Accordingly, I hypothesized that the PPT GABA(B) receptor activation-mediated REM sleep suppression effect could be mediated through inhibition of cAMP-PKA activation. To test this hypothesis, a GABA(B) receptor selective agonist, baclofen hydrochloride (baclofen), cAMP-PKA activator, Sp-adenosine 3',5'-cyclic monophosphothioate triethylamine (SpCAMPS), and vehicle control were microinjected into the PPT in selected combinations to determine effects on sleep-waking states of chronically instrumented, freely moving rats. Microinjection of SpCAMPS (1.5 nmol) induced REM sleep within a short latency (12.1 +/- 3.6 min) compared with vehicle control microinjection (60.0 +/- 6.5 min). On the contrary, microinjection of baclofen (1.5 nmol) suppressed REM sleep by delaying its appearance for approximately 183 min; however, the suppression of REM sleep by baclofen was prevented by a subsequent microinjection of SpCAMPS. These results provide evidence that the activation of cAMP-PKA within the PPT can successfully block the GABA(B) receptor activation mediated REM sleep suppression effect. These findings suggest that the PPT GABA(B) receptor activation-mediated REM sleep regulating mechanism involves inactivation of cAMP-PKA signaling in the freely moving rat. PMID- 17409166 TI - Self-tuning of neural circuits through short-term synaptic plasticity. AB - Numerous experimental data show that cortical networks of neurons are not silent in the absence of external inputs, but rather maintain a low spontaneous firing activity. This aspect of cortical networks is likely to be important for their computational function, but is hard to reproduce in models of cortical circuits of neurons because the low-activity regime is inherently unstable. Here we show through theoretical analysis and extensive computer simulations-that short-term synaptic plasticity endows models of cortical circuits with a remarkable stability in the low-activity regime. This short-term plasticity works as a homeostatic mechanism that stabilizes the overall activity level in spite of drastic changes in external inputs and internal circuit properties, while preserving reliable transient responses to signals. The contribution of synaptic dynamics to this stability can be predicted on the basis of general principles from control theory. PMID- 17409167 TI - Mathematical model of network dynamics governing mouse sleep-wake behavior. AB - Recent work in experimental neurophysiology has identified distinct neuronal populations in the rodent brain stem and hypothalamus that selectively promote wake and sleep. Mutual inhibition between these cell groups has suggested the conceptual model of a sleep-wake switch that controls transitions between wake and sleep while minimizing time spent in intermediate states. By combining wake- and sleep-active populations with populations governing transitions between different stages of sleep, a "sleep-wake network" of neuronal populations may be defined. To better understand the dynamics inherent in this network, we created a model sleep-wake network composed of coupled relaxation oscillation equations. Mathematical analysis of the deterministic model provides insight into the dynamics underlying state transitions and predicts mechanisms for each transition type. With the addition of noise, the simulated sleep-wake behavior generated by the model reproduces many qualitative and quantitative features of mouse sleep wake behavior. In particular, the existence of simulated brief awakenings is a unique feature of the model. In addition to capturing the experimentally observed qualitative difference between brief and sustained wake bouts, the model suggests distinct network mechanisms for the two types of wakefulness. Because circadian and other factors alter the fine architecture of sleep-wake behavior, this model provides a novel framework to explore dynamical principles that may underlie normal and pathologic sleep-wake physiology. PMID- 17409168 TI - Enhancement of visual responsiveness by spontaneous local network activity in vivo. AB - Spontaneous activity within local circuits affects the integrative properties of neurons and networks. We have previously shown that neocortical network activity exhibits a balance between excitatory and inhibitory synaptic potentials, and such activity has significant effects on synaptic transmission, action potential generation, and spike timing. However, whether such activity facilitates or reduces sensory responses has yet to be clearly determined. We examined this hypothesis in the primary visual cortex in vivo during slow oscillations in ketamine-xylazine anesthetized cats. We measured network activity (Up states) with extracellular recording, while simultaneously recording postsynaptic potentials (PSPs) and action potentials in nearby cells. Stimulating the receptive field revealed that spiking responses of both simple and complex cells were significantly enhanced (>2-fold) during network activity, as were spiking responses to intracellular injection of varying amplitude artificial conductance stimuli. Visually evoked PSPs were not significantly different in amplitude during network activity or quiescence; instead, spontaneous depolarization caused by network activity brought these evoked PSPs closer to firing threshold. Further examination revealed that visual responsiveness was gradually enhanced by progressive membrane potential depolarization. These spontaneous depolarizations enhanced responsiveness to stimuli of varying contrasts, resulting in an upward (multiplicative) scaling of the contrast response function. Our results suggest that small increases in ongoing balanced network activity that result in depolarization may provide a rapid and generalized mechanism to control the responsiveness (gain) of cortical neurons, such as occurs during shifts in spatial attention. PMID- 17409169 TI - Factors affecting the common modulation of bilateral motor unit discharge in human soleus muscles. AB - The purpose of this study was to determine the factors that influence the co modulation of motor unit discharge rate in soleus muscles of both legs during upright standing. Single motor units were recorded from the left and right soleus muscles under three experimental conditions: standing quietly with the eyes open and closed, standing with the eyes closed while vibration was applied to one Achilles tendon, and swaying voluntarily or producing variable low-force isometric contractions at a frequency of 0.05 Hz. Correlations in motor unit discharge rate between left and right soleus motor units were assessed using common drive analysis. The results showed that common drive to motoneurons of the two muscles did not differ between standing with the eyes open or closed, but there was an order effect with the second task having significantly lower common drive than the first. Common drive was also significantly lower when vibration was applied to one leg compared with when no vibration was applied. Common drive was higher as subjects swayed anteriorly as compared with when they swayed posteriorly. There were no significant differences in common drive across phases of the variable isometric force contraction. Common drive was higher during voluntary sway than during variable force production; both of these values were significantly lower than those derived from the quiet standing task. These results suggest that proprioceptive and sub-cortical inputs contribute to the co modulation of the firing rate of soleus motor unit pairs of the left and right leg during standing posture. PMID- 17409170 TI - Panulirus interruptus Ih-channel gene PIIH: modification of channel properties by alternative splicing and role in rhythmic activity. AB - We cloned 10 full-length variants of PIIH, the gene for I(h) from the spiny lobster, Panulirus interruptus, using reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). This gene shows a significant amount of alternative splicing in the S3-S4 and S4-S5 linkers, in the P-loop and the entire S6 transmembrane domain, in the cyclic nucleotide binding domain (CNBD), and near the 3' end of the gene. Functional expression of seven splice variants in Xenopus oocytes generated slowly activating hyperpolarization-activated inward currents, which were blocked by the I(h) channel blockers CsCl and ZD7288. The different splice variants had markedly varying activation kinetics and voltage dependence of activation. Bath application of 8-Br-cAMP shifted the V(1/2) to more positive potentials and accelerated the activation kinetics in an isoform-specific manner. Two variants containing a segment with an ER-retention motif in the S4-S5 loop did not produce currents in oocytes. Overexpression of one splice variant, PIIH AB(S)-I, in pyloric dilator (PD) neurons in the lobster stomatogastric ganglion produced an average threefold increase in I(h) without evoking a compensatory increase in I(A). The voltage for half-maximal activation of I(h) in PIIH AB(S)-I expressing PDs was shifted in the depolarizing direction by 9 mV, whereas the slope factor decreased by 3.8 mV. Moreover, its activation kinetics were significantly faster than in control PDs. PIIH AB(S)-I overexpression enhanced PD neuron rhythmic firing in an amplitude-dependent manner above a minimal threshold two- to threefold increase in amplitude. PMID- 17409171 TI - Cortical spreading depression--better understanding and more questions. Focus on "distinct vascular conduction with cortical spreading depression". PMID- 17409172 TI - Neurokinin receptor-expressing pre-botzinger complex neurons in neonatal mice studied in vitro. AB - Breathing in mammals depends on inspiratory-related neural activity generated in the pre-Botzinger complex (preBotC), where neurokinin receptor-expressing neurons (NKR(+)) have been hypothesized to play a critical rhythmogenic role. Currently, the extent to which the preBotC is populated by rhythmogenic NKR(+) neurons and whether neurons without neurokinin receptor expression (NKR(-)) share similar electrical properties with NKR(+) neurons are not well understood. These interrelated problems must be resolved to understand the widespread excitatory effects of neuropeptides and the mechanism of respiratory rhythmogenesis. We recorded and imaged inspiratory neurons in neonatal mouse slices that isolate the preBotC and generate respiratory motor output in vitro. Using tetramethylrhodamine conjugated to the endogenous NKR agonist substance P (TMR SP) to tag neurons that express NKRs, we show that early inspiratory neurons with small whole cell capacitance (C(M)) are 36% TMR-SP(+) and 64% TMR-SP(-). Also, late inspiratory neurons with large C(M) are 67% TMR-SP(+) and 33% are TMR-SP(-). Thus NKR(+) and NKR(-) neurons exhibit the same phenotypic properties, which suggests that they may share functional roles also. Substance P (SP) alone evoked a voltage-insensitive inward current (I(SP)) that reversed at -19 mV and was associated with an increase in membrane conductance in both NKR(+) and NKR(-) neurons. Gap junctions may be needed to confer SP sensitivity to neurons that appear to lack NKR expression. We propose that cell death in NKR(+) preBotC neurons, by targeted lesion or neurodegeneration, may impair breathing behavior by killing less than one half of the rhythmogenic preBotC neurons and a large number of respiratory premotoneurons. PMID- 17409173 TI - Activity-dependent depression of local excitatory connections in the CA1 region of mouse hippocampus. AB - The existence of recurrent excitatory synapses between pyramidal cells in the hippocampal CA1 region has been known for some time yet little is known about activity-dependent forms of plasticity at these synapses. Here we demonstrate that under certain experimental conditions, Schaffer collateral/commissural fiber stimulation can elicit robust polysynaptic excitatory postsynaptic potentials due to recurrent synaptic inputs onto CA1 pyramidal cells. In contrast to CA3 pyramidal cell inputs, recurrent synapses onto CA1 pyramidal cells exhibited robust paired-pulse depression and a sustained, but rapidly reversible, depression in response to low-frequency trains of Schaffer collateral fiber stimulation. Blocking GABA(B) receptors abolished paired-pulse depression but had little effect on low-frequency stimulation (LFS)-induced depression. Instead, LFS induced depression was significantly attenuated by an inhibitor of A1 type adenosine receptors. Blocking the postsynaptic effects of GABA(B) and A1 receptor activation on CA1 pyramidal cell excitability with an inhibitor of G-protein activated inwardly rectifying potassium channels had no effect on either paired pulse depression or LFS-induced depression. Thus activation of presynaptic GABA(B) and adenosine receptors appears to have an important role in activity dependent depression at recurrent synapses. Together, our results indicate that CA3-CA1 and CA1-CA1 synapses exhibit strikingly different forms of short-term synaptic plasticity and suggest that activity-dependent changes in recurrent synaptic transmission can transform the CA1 region from a sparsely connected recurrent network into a predominantly feedforward circuit. PMID- 17409174 TI - Impact of coordinate transformation uncertainty on human sensorimotor control. AB - Humans build representations of objects and their locations by integrating imperfect information from multiple perceptual modalities (e.g., visual, haptic). Because sensory information is specified in different frames of reference (i.e., eye- and body-centered), it must be remapped into a common coordinate frame before integration and storage in memory. Such transformations require an understanding of body articulation, which is estimated through noisy sensory data. Consequently, target information acquires additional coordinate transformation uncertainty (CTU) during remapping because of errors in joint angle sensing. As a result, CTU creates differences in the reliability of target information depending on the reference frame used for storage. This paper explores whether the brain represents and compensates for CTU when making grasping movements. To address this question, we varied eye position in the head, while participants reached to grasp a spatially fixed object, both when the object was in view and when it was occluded. Varying eye position changes CTU between eye and head, producing additional uncertainty in remapped information away from forward view. The results showed that people adjust their maximum grip aperture to compensate both for changes in visual information and for changes in CTU when the target is occluded. Moreover, the amount of compensation is predicted by a Bayesian model for location inference that uses eye-centered storage. PMID- 17409175 TI - Synaptic amplification versus bistability in motoneuron dendritic processing: a top-down modeling approach. AB - Motoneurons have been shown to exhibit both bistable firing and synaptic amplification. Both of these behaviors have generally been attributed to a single mechanism-dendritic plateau potentials based on L-type Ca(2+) conductances. However, our recent discovery of a fast-amplification mode calls this into question. Here we examine the possibility that two mechanisms underlie these behaviors, one being a slow-mode bistability mechanism (i.e., the L-type Ca(2+) conductance-based dendritic plateaus) and the other being a theoretical fast-mode amplification mechanism. A "top-down" motoneuron model that encapsulated these and other hypotheses was developed in which these mechanisms could be explored. The resulting final model simultaneously exhibits synaptic amplification, plateau potential formation, bistable firing patterns, and current-voltage (I-V) and frequency-current (F-I) hystereses. This model suggests that amplification and plateaus are mutually exclusive in the same dendrite/dendritic branch. Thus we predict that plateau generation does not occur in all dendritic branches. This could be readily accomplished by a large degree of variation in the density of L type Ca(2+) channels believed to underlie plateau formation in these cells with the added benefit of spreading plateau onset over a wider voltage range, as is observed experimentally. PMID- 17409176 TI - Interval coding. I. Burst interspike intervals as indicators of stimulus intensity. AB - Short interspike intervals such as those that occur during burst firing are hypothesized to be distinct features of the neural code. Although a number of correlations between the occurrence of burst events and aspects of the stimulus have been identified, the relationship between burst characteristics and information transfer is uncertain. Pyramidal cells in the electrosensory lobe of the weakly electric fish, Apteronotus leptorhynchus, respond to dynamic broadband electrosensory stimuli with bursts and isolated spikes. In the present study, we mimic synaptic input during sensory stimulation by direct stimulation of electrosensory pyramidal cells with broadband current in vitro. The pyramidal cells respond to this stimulus with burst interspike intervals (ISIs) that are reliably and precisely correlated with the intensity of stimulus upstrokes. We found burst ISIs must differ by a minimum of 2 ms to discriminate, with low error, differences in stimulus intensity. Based on these results, we define and quantify a candidate interval code for the processing of sensory input. Finally, we demonstrate that interval coding is restricted to short ISIs such as those generated in burst events and that the proposed interval code is distinct from rate and timing codes. PMID- 17409177 TI - Interval coding. II. Dendrite-dependent mechanisms. AB - The rich temporal structure of neural spike trains provides multiple dimensions to code dynamic stimuli. Popular examples are spike trains from sensory cells where bursts and isolated spikes can serve distinct coding roles. In contrast to analyses of neural coding, the cellular mechanics of burst mechanisms are typically elucidated from the neural response to static input. Bridging the mechanics of bursting with coding of dynamic stimuli is an important step in establishing theories of neural coding. Electrosensory lateral line lobe (ELL) pyramidal neurons respond to static inputs with a complex dendrite-dependent burst mechanism. Here we show that in response to dynamic broadband stimuli, these bursts lack some of the electrophysiological characteristics observed in response to static inputs. A simple leaky integrate-and-fire (LIF)-style model with a dendrite-dependent depolarizing afterpotential (DAP) is sufficient to match both the output statistics and coding performance of experimental spike trains. We use this model to investigate a simplification of interval coding where the burst interspike interval (ISI) codes for the scale of a canonical upstroke rather than a multidimensional stimulus feature. Using this stimulus reduction, we compute a quantization of the burst ISIs and the upstroke scale to show that the mutual information rate of the interval code is maximized at a moderate DAP amplitude. The combination of a reduced description of ELL pyramidal cell bursting and a simplification of the interval code increases the generality of ELL burst codes to other sensory modalities. PMID- 17409178 TI - Clinical epidemiological study of employees exposed to surfactant blend containing perfluorononanoic acid. AB - INTRODUCTION: An epidemiological study was conducted of a perfluorononanoic acid (PFNA) surfactant blend, to investigate whether clinical differences were apparent between employees who were potentially exposed to the surfactant and those who were not exposed. The surfactant blend, which is related to other previously studied perfluorinated materials, is used in the production of some high-performance polymers. METHODS: All 630 individuals employed at a polymer production facility using PFNA (CAS No 72968-38-8) at any time between 1 January 1989 and 1 July 2003 were included in the cohort. Plausibly related laboratory test results were abstracted from annual medical examination records, including liver enzyme function and blood lipids. Detailed work histories, available for all employees, provided the basis for determining exposure category. Thirty two clinical parameters were evaluated by exposure level at five points in time, determined to reflect changes in possible exposure intensity, as well as greatest number of records available. Annual cross-sectional analyses and longitudinal analyses that accounted for multiple measurements per person were conducted separately for men and women, by exposure groups. RESULTS: Differences by exposure group for all laboratory measures, adjusted for age and body mass index, were small and not clinically significant. Although some statistically significant pair-wise differences were observed, these observations were not consistent between men and women, or over the five analysis windows. For the seven outcome variables (liver enzymes and blood lipids) examined in separate longitudinal models, no significant increase or decrease was observed by unit increase in cumulative exposure intensity score. CONCLUSION: This is the first epidemiological study investigating the possible health effects in humans associated with exposure to PFNA blend. Based on laboratory measures assessed over more than a decade, no adverse clinical effects were detected from occupational exposure to PFNA blend. PMID- 17409179 TI - Long-term use of cellular phones and brain tumours: increased risk associated with use for > or =10 years. AB - AIM: To evaluate brain tumour risk among long-term users of cellular telephones. METHODS: Two cohort studies and 16 case-control studies on this topic were identified. Data were scrutinised for use of mobile phone for > or =10 years and ipsilateral exposure if presented. RESULTS: The cohort study was of limited value due to methodological shortcomings in the study. Of the 16 case-control studies, 11 gave results for > or =10 years' use or latency period. Most of these results were based on low numbers. An association with acoustic neuroma was found in four studies in the group with at least 10 years' use of a mobile phone. No risk was found in one study, but the tumour size was significantly larger among users. Six studies gave results for malignant brain tumours in that latency group. All gave increased odd ratios (OR), especially for ipsilateral exposure. In a meta analysis, ipsilateral cell phone use for acoustic neuroma was OR = 2.4 (95% CI 1.1 to 5.3) and OR = 2.0, (1.2 to 3.4) for glioma using a tumour latency period of > or =10 years. CONCLUSIONS: Results from present studies on use of mobile phones for > or =10 years give a consistent pattern of increased risk for acoustic neuroma and glioma. The risk is highest for ipsilateral exposure. PMID- 17409180 TI - The effect of individual counselling and education on work ability and disability pension: a prospective intervention study in the construction industry. AB - OBJECTIVES: To investigate the effectiveness of a counselling and education programme on work ability and work disability pension for employees in the construction industry. METHODS: Employees with a high disability risk of 38% or more in the following four years were included. Employees in the intervention group were either selected by an occupational physician or enrolled themselves. They received an assessment and individual programme focused on optimising work functioning, while the control group received care-as-usual. Data on work ability measured with the Work Ability Index (WAI) and work disability pensions were collected at baseline during a periodic occupational health examination and at 9, 18 and 26 months after the start of the intervention using a questionnaire. RESULTS: Most employees in both the intervention (n = 83) and control group (n = 209) were carpenters (43% and 37%) and bricklayers (7% and 15%). In the intervention group, 42% successfully completed the programme. Work ability in the intervention group was lower at baseline but showed an increase over time while work ability of the control group remained the same. The work ability in the intervention group improved slightly more (p = 0.09). No statistically significant differences in percentages of employees receiving a disability pension between the intervention and control group were found at 9 or 18 months and no differences in the age-adjusted percentages of employees receiving a disability pension were found between the groups at any measurement. CONCLUSIONS: The programme was slightly effective in improving the work ability but not in reducing work disability pensions. A more comprehensive multidisciplinary intervention programme might be necessary. PMID- 17409181 TI - Short-term effects of ozone air pollution on ischaemic stroke occurrence: a case crossover analysis from a 10-year population-based study in Dijon, France. AB - OBJECTIVE: To evaluate the association between air pollutants and the occurrence of acute stroke from 10-year population-based study. METHODS: The daily stroke count was obtained from Dijon Stroke Register between March 1994 and December 2004. The register recorded all first-ever strokes among residents of Dijon (150,000 inhabitants) in France, using standard diagnostic criteria. Pollutant concentrations (SO2, CO, NO2, O3 and PM10) were measured hourly. A bi-directional case-crossover design was used to examine the association between air pollutant and stroke onset. The conditional logistic regression model included the meteorological parameters (temperature, relative humidity), influenza epidemics and holidays. RESULTS: The authors collected 493 large artery infarcts, 397 small artery infarcts, 530 cardio-embolic infarcts, 67 undeterminate infarcts, 371 transient ischaemic attacks and 220 haemorrhagic strokes. For single-pollutant model and for a 10 mg/m(3) increase of O3 exposure, a positive association was observed only in men, over 40 years of age, between ischaemic stroke occurrence and O3 levels with 1-day lag, (OR 1.133, 95% CI 1.052 to 1.220) and 0-day lag (OR 1.058, 95% CI 0.987 to 1.134). No significant associations were found for haemorrhagic stroke. In two-pollutant models, the effects of O3 remained significant after each of the other pollutants were included in the model, in particular with PM10. A significant association was observed for ischaemic strokes of large arteries (p = 0.02) and for transient ischaemic attacks (p = 0.01). Moreover, the authors found an exposure-response relations between O3 exposure and ischaemic stroke (test for trend, p = 0.01). An increase in association in men with several cardiovascular risk factors (smoker, dyslipidemia and hypertension) was also observed. CONCLUSION: These observational data argue for an association between ischaemic stroke occurrence and O3 pollution levels; these results still need to be confirmed by other studies. PMID- 17409182 TI - The pro-inflammatory effects of low-toxicity low-solubility particles, nanoparticles and fine particles, on epithelial cells in vitro: the role of surface area. AB - OBJECTIVE: Rats exposed to high airborne mass concentrations of low-solubility low-toxicity particles (LSLTP) have been reported to develop lung disease such as fibrosis and lung cancer. These particles are regulated on a mass basis in occupational settings, but mass might not be the appropriate metric as animal studies have shown that nanoparticles (ultrafine particles) produce a stronger adverse effect than fine particles when delivered on an equal mass basis. METHODS: This study investigated whether the surface area is a better descriptor than mass of LSLTP of their ability to stimulate pro-inflammatory responses in vitro. In a human alveolar epithelial type II-like cell line, A549, we measured interleukin (IL)-8 mRNA, IL8 protein release and glutathione (GSH) depletion as markers of pro-inflammatory effects and oxidative stress after treatment with a range of LSLTP (fine and nanoparticles) and DQ12 quartz, a particle with a highly reactive surface. RESULTS: In all the assays, nanoparticle preparations of titanium dioxide (TiO2-np) and of carbon black (CB-np) produced much stronger pro inflammatory responses than the same mass dose of fine TiO2 and CB. The results of the GSH assay confirmed that oxidative stress was involved in the response to all the particles, and two ultra-fine metal dusts (cobalt and nickel) produced GSH depletion similar to TiO2-np, for similar surface-area dose. As expected, DQ12 quartz was more inflammatory than the low toxicity dusts, on both a mass and surface-area basis. CONCLUSION: Dose-response relationships observed in the in vitro assays appeared to be directly comparable with dose-response relationships in vivo when the doses were similarly standardised. Both sets of data suggested a threshold in dose measured as surface area of particles relative to the surface area of the exposed cells, at around 1-10 cm2/cm2. These findings are consistent with the hypothesis that surface area is a more appropriate dose metric than mass for the pro-inflammatory effects of LSLTP in vitro and in vivo, and consequently that the high surface area of nanoparticles is a key factor in their inflammogenicity. PMID- 17409183 TI - A simple diagnostic model for ruling out pneumoconiosis among construction workers. AB - BACKGROUND: Construction workers exposed to silica-containing dust are at risk of developing silicosis even at low exposure levels. Health surveillance among these workers is commonly advised but the exact diagnostic work-up is not specified and therefore may result in unnecessary chest x ray investigations. AIM: To develop a simple diagnostic model to estimate the probability of an individual worker having pneumoconiosis from questionnaire and spirometry results, in order to accurately rule out workers without pneumoconiosis. METHODS: The study was performed using cross-sectional data of 1291 Dutch natural stone and construction workers with potentially high quartz dust exposure. A multivariable logistic regression model was developed using chest x ray with ILO profusion category > or =1/1 as the reference standard. The model's calibration was evaluated with the Hosmer-Lemeshow test; the discriminative ability was determined by calculating the area under the receiver operating characteristic curve (ROC area). Internal validity of the final model was assessed by a bootstrapping procedure. For clinical application, the diagnostic model was transformed into an easy-to-use score chart. RESULTS: Age 40 years or older, current smoker, high-exposure job, working 15 years or longer in the construction industry, "feeling unhealthy" and FEV1 were independent predictors in the diagnostic model. The model showed good calibration (a non-significant Hosmer-Lemeshow test) and discriminative ability (ROC area 0.81, 95% CI 0.74 to 0.85). Internal validity was reasonable; the optimism corrected ROC area was 0.76. By using a cut-off point with a high negative predictive value the occupational physician can efficiently detect a large proportion of workers with a low probability of having pneumoconiosis and exclude them from unnecessary x ray investigations. CONCLUSIONS: This diagnostic model is an efficient and effective instrument to rule out pneumoconiosis among construction workers. Its use in health surveillance among these workers can reduce the number of redundant x ray investigations. PMID- 17409184 TI - Phase I safety and pharmacokinetic study of magnesium isoglycyrrhizinate after single and multiple intravenous doses in chinese healthy volunteers. AB - The safety and pharmacokinetics of magnesium isoglycyrrhizinate were assessed in healthy Chinese volunteers. In the single-dose format of this pharmacokinetic study, 100-, 200-, and 300-mg doses of magnesium isoglycyrrhizinate were given by intravenous infusion. The results indicated that the plasma levels were directly proportional to the administered dose, with the mean C(max) and AUC(0-72) ranging from approximately 28.79 to 99.28 mg x L(-1) and 448.68 to 1688.42 mg x h x L(-1) over the dose range. In the multiple-dose format of this pharmacokinetic study, 100 mg magnesium isoglycyrrhizinate was administrated once daily for 9 days. Moderate drug accumulation was noted, which was attributable to the drug's long terminal half-life of 19 to 31 hours. The distribution and elimination rate of magnesium isoglycyrrhizinate had no changes. It had a favorable pharmacokinetics and safety profile that enables the drug to be explored in future clinical studies that target patients with hepatic impairment. PMID- 17409185 TI - DNA demethylation in the Arabidopsis genome. AB - Cytosine DNA methylation is considered to be a stable epigenetic mark, but active demethylation has been observed in both plants and animals. In Arabidopsis thaliana, DNA glycosylases of the DEMETER (DME) family remove methylcytosines from DNA. Demethylation by DME is necessary for genomic imprinting, and demethylation by a related protein, REPRESSOR OF SILENCING1, prevents gene silencing in a transgenic background. However, the extent and function of demethylation by DEMETER-LIKE (DML) proteins in WT plants is not known. Using genome-tiling microarrays, we mapped DNA methylation in mutant and WT plants and identified 179 loci actively demethylated by DML enzymes. Mutations in DML genes lead to locus-specific DNA hypermethylation. Reintroducing WT DML genes restores most loci to the normal pattern of methylation, although at some loci, hypermethylated epialleles persist. Of loci demethylated by DML enzymes, >80% are near or overlap genes. Genic demethylation by DML enzymes primarily occurs at the 5' and 3' ends, a pattern opposite to the overall distribution of WT DNA methylation. Our results show that demethylation by DML DNA glycosylases edits the patterns of DNA methylation within the Arabidopsis genome to protect genes from potentially deleterious methylation. PMID- 17409186 TI - Prevalence of positive selection among nearly neutral amino acid replacements in Drosophila. AB - We have estimated the selective effects of amino acid replacements in natural populations by comparing levels of polymorphism in 91 genes in African populations of Drosophila melanogaster with their divergence from Drosophila simulans. The genes include about equal numbers whose level of expression in adults is greater in males, greater in females, or approximately equal in the sexes. Markov chain Monte Carlo methods were used to sample key parameters in the stationary distribution of polymorphism and divergence in a model in which the selective effect of each nonsynonymous mutation is regarded as a random sample from some underlying normal distribution whose mean may differ from one gene to the next. Our analysis suggests that approximately 95% of all nonsynonymous mutations that could contribute to polymorphism or divergence are deleterious, and that the average proportion of deleterious amino acid polymorphisms in samples is approximately 70%. On the other hand, approximately 95% of fixed differences between species are positively selected, although the scaled selection coefficient (N(e)s) is very small. We estimate that approximately 46% of amino acid replacements have N(e)s < 2, approximately 84% have N(e)s < 4, and approximately 99% have N(e)s < 7. Although positive selection among amino acid differences between species seems pervasive, most of the selective effects could be regarded as nearly neutral. There are significant differences in selection between sex-biased and unbiased genes, which relate primarily to the mean of the distributions of mutational effects and the fraction of slightly deleterious and weakly beneficial mutations that are fixed. PMID- 17409187 TI - Caspase 1 deficiency reduces inflammation-induced brain transcription. AB - The systemic inflammatory response syndrome (SIRS) is a life-threatening medical condition characterized by a severe and generalized inflammatory state that can lead to multiple organ failure and shock. The CNS regulates many features of SIRS such as fever, cardiovascular, and neuroendocrine responses. Central and systemic manifestations of SIRS can be induced by LPS or IL-1beta administration. The crucial role of IL-1beta in inflammation has been further highlighted by studies of mice lacking caspase 1 (casp1, also known as IL-1beta convertase), a protease that cleaves pro-IL-1beta into mature IL-1beta. Indeed, casp1 knockout (casp1(-/ )) mice survive lethal doses of LPS. The key role of IL-1beta in sickness behavior and its de novo expression in the CNS during inflammation led us to test the hypothesis that IL-1beta plays a major role modulating the brain transcriptome during SIRS. We show a gene-environment effect caused by LPS administration in casp1(-/-) mice. During SIRS, the expression of several genes, such as chemokines, GTPases, the metalloprotease ADAMTS1, IL-1RA, the inducible nitric oxide synthase, and cyclooxygenase-2, was differentially increased in casp1(-/-) mice. Our findings may contribute to the understanding of the molecular changes that take place within the CNS during sepsis and SIRS and the development of new therapies for these serious conditions. Our results indicate that those genes may also play a role in several neuropsychiatric conditions in which inflammation has been implicated and indicate that casp1 might be a potential therapeutic target for such disorders. PMID- 17409188 TI - Role for Msh5 in the regulation of Ig class switch recombination. AB - Ig class switch recombination (CSR) and somatic hypermutation serve to diversify antibody responses and are orchestrated by the activity of activation-induced cytidine deaminase and many proteins involved in DNA repair and genome surveillance. Msh5, a gene encoded in the central MHC class III region, and its obligate heterodimerization partner Msh4 have a critical role in regulating meiotic homologous recombination and have not been implicated in CSR. Here, we show that MRL/lpr mice carrying a congenic H-2(b/b) MHC interval exhibit several abnormalities regarding CSR, including a profound deficiency of IgG3 in most mice and long microhomologies at Ig switch (S) joints. We found that Msh5 is expressed at low levels on the H-2(b) haplotype and, importantly, a similar long S joint microhomology phenotype was observed in both Msh5 and Msh4-null mice. We also present evidence that genetic variation in MSH5 is associated with IgA deficiency and common variable immune deficiency (CVID) in humans. One of the human MSH5 alleles identified contains two nonsynonymous polymorphisms, and the variant protein encoded by this allele shows impaired binding to MSH4. Similar to the mice, Ig S joints from CVID and IgA deficiency patients carrying disease associated MSH5 alleles show increased donor/acceptor microhomology, involving pentameric DNA repeat sequences and lower mutation rates than controls. Our findings suggest that Msh4/5 heterodimers contribute to CSR and support a model whereby Msh4/5 promotes the resolution of DNA breaks with low or no terminal microhomology by a classical nonhomologous end-joining mechanism while possibly suppressing an alternative microhomology-mediated pathway. PMID- 17409189 TI - Clustering of peptidoglycan recognition protein-SA is required for sensing lysine type peptidoglycan in insects. AB - Recognition of lysine-type peptidoglycan by peptidoglycan recognition protein (PGRP)-SA provokes the activation of the Toll and prophenoloxidase pathways. Here we reveal that a soluble fragment of lysine-type peptidoglycan, a long glycan chain with short stem peptides, is a potent activator of the Drosophila Toll pathway and the prophenoloxidase activation cascade in the beetle Tenebrio molitor. Using this peptidoglycan fragment, we present biochemical evidence that clustering of PGRP-SA molecules on the peptidoglycan is required for the activation of the prophenoloxidase cascade. We subsequently highlight that the lysozyme-mediated partial digestion of highly cross-linked lysine-type peptidoglycan dramatically increases the binding of PGRP-SA, presumably by inducing clustering of PGRP-SA, which then recruits the Gram-negative bacteria binding protein 1 homologue and a modular serine protease containing low-density lipoprotein and complement control protein domains. The crucial role of lysozyme in the prophenoloxidase activation cascade is further confirmed in vivo by using a lysozyme inhibitor. Taken together, we propose a model whereby lysozyme presents a processed form of lysine-type peptidoglycan for clustering of PGRP-SA that recruits Gram-negative bacteria-binding protein 1 and the modular serine protease, which leads to the activation of both the Toll and prophenoloxidase pathways. PMID- 17409190 TI - Time-resolved infrared (TRIR) study on the formation and reactivity of organometallic methane and ethane complexes in room temperature solution. AB - We have used fast time-resolved infrared spectroscopy to characterize a series of organometallic methane and ethane complexes in solution at room temperature: W(CO)5(CH4) and M(eta5-C5R5)(CO)2(L) [where M = Mn or Re, R = H or CH3 (Re only); and L = CH4 or C2H6]. In all cases, the methane complexes are found to be short lived and significantly more reactive than the analogous n-heptane complexes. Re(Cp)(CO)2(CH4) and Re(Cp*)(CO)2(L) [Cp* = eta5-C5(CH3)(5) and L = CH4, C2H6] were found to be in rapid equilibrium with the alkyl hydride complexes. In the presence of CO, both alkane and alkyl hydride complexes decay at the same rate. We have used picosecond time-resolved infrared spectroscopy to directly monitor the photolysis of Re(Cp*)(CO)3 in scCH4 and demonstrated that the initially generated Re(Cp*)(CO)2(CH4) forms an equilibrium mixture of Re(Cp*)(CO)2(CH4)/Re(Cp*)(CO)2(CH3)H within the first few nanoseconds (tau = 2 ns). The ratio of alkane to alkyl hydride complexes varies in the order Re(Cp)(CO)2(C2H6):Re(Cp)(CO)2(C2H5)H > Re(Cp*)(CO)2(C2H6):Re(Cp*)(CO)2(C2H5)H approximately equal to Re(Cp)(CO)2(CH4):Re(Cp)(CO)2(CH3)H > Re(Cp*)(CO)2(CH4):Re(Cp*)(CO)2(CH3)H. Activation parameters for the reactions of the organometallic methane and ethane complexes with CO have been measured, and the DeltaH++ values represent lower limits for the CH4 binding enthalpies to the metal center of W-CH4 (30 kJ.mol(-1)), Mn-CH4 (39 kJ.mol(-1)), and Re-CH4 (51 kJ.mol(-1)) bonds in W(CO)5(CH4), Mn(Cp)(CO)2(CH4), and Re(Cp)(CO)2(CH4), respectively. PMID- 17409191 TI - Indian Hedgehog produced by postnatal chondrocytes is essential for maintaining a growth plate and trabecular bone. AB - Indian hedgehog (Ihh) is essential for chondrocyte and osteoblast proliferation/differentiation during prenatal endochondral bone formation. The early lethality of various Ihh-ablated mutant mice, however, prevented further analysis of its role in postnatal bone growth and development. In this study, we describe the generation and characterization of a mouse model in which the Ihh gene was successfully ablated from postnatal chondrocytes in a temporal/spatial specific manner; postnatal deletion of Ihh resulted in loss of columnar structure, premature vascular invasion, and formation of ectopic hypertrophic chondrocytes in the growth plate. Furthermore, destruction of the articular surface in long bones and premature fusion of growth plates of various endochondral bones was evident, resulting in dwarfism in mutant mice. More importantly, these mutant mice exhibited continuous loss of trabecular bone over time, which was accompanied by reduced Wnt signaling in the osteoblastic cells. These results demonstrate, for the first time, that postnatal chondrocyte-derived Ihh is essential for maintaining the growth plate and articular surface and is required for sustaining trabecular bone and skeletal growth. PMID- 17409192 TI - Genomewide suggestive linkage of opioid dependence to chromosome 14q. AB - The genetic predisposition to addiction to opioids and other substances is transmitted as a complex genetic trait, which investigators are attempting to characterize using genetic linkage and association. We now report a high-density genome-wide linkage study of opioid dependence. We ascertained 305 DSM-IV opioid dependent affected sibling pairs from an ethnically mixed population of methadone maintained subjects and genotyped their DNA using Affymetrix 10K v2 arrays. Analysis with MERLIN identified a region on chromosome 14q with a non-parametric lod (NPL) of 3.30. Secondary analyses indicated that this locus was relatively specific to the self-identified Puerto Rican subset, as the NPL increased from 3.30 to 5.00 (NPL(Caucasian) = 0.05 and NPL(African Amer.) = 0.15). The 14q peak encompasses the NRXN3 gene (neurexin 3), which was previously identified as a potential candidate gene for addiction. Secondary analyses also identified several regions with gender-specific NPL scores greater than 2.00. The most significant was a peak on (10q) that increased from 0.90 to 3.22 when only males were considered (NPL(female) = 0.05). Our linkage data suggest specific chromosomal loci for future fine-mapping genetic analysis and support the hypothesis that ethnic and gender specific genes underlie addiction susceptibility. PMID- 17409193 TI - Apoptotic mechanisms in mutant LRRK2-mediated cell death. AB - Mutations in the gene coding for leucine-rich repeat kinase 2 (LRRK2) cause autosomal-dominant Parkinson's disease. The pathological mutations have been associated with an increase of LRRK2 kinase activity, although its physiological substrates have not been identified yet. The data we report here demonstrate that disease-associated mutant LRRK2 cell toxicity is due to mitochondria-dependent apoptosis. Transient transfection of mutant LRRK2 leads to neuronal death with clear apoptotic signs. Soluble caspase inhibitors or the genetic ablation of Apaf1 protects cells from apoptotic death. Moreover, we explored the function of two protein domains in LRRK2 (LRR and WD40) and demonstrate that the lack of these protein domains has a protective effect on mitochondria dysfunctions induced by mutant LRRK2. PMID- 17409194 TI - Histones associated with downregulated genes are hypo-acetylated in Huntington's disease models. AB - Transcriptional dysregulation plays a major role in the pathology of Huntington's disease (HD). However, the mechanisms causing selective downregulation of genes remain unknown. Histones regulate chromatin structure and thereby control gene expression; recent studies have demonstrated a therapeutic role for histone deacetylase (HDAC) inhibitors in polyglutamine diseases. This study demonstrates that despite no change in overall acetylated histone levels, histone H3 is hypo acetylated at promoters of downregulated genes in R6/2 mice, ST14a and STHdh cells, as demonstrated by in vivo chromatin immunoprecipitation. In addition, HDAC inhibitor treatment increases association of acetylated histones with downregulated genes and corrects mRNA abnormalities. In contrast, there is a decrease in mRNA levels in wild-type cells following treatment with a histone acetyltransferase inhibitor. Although changes in histone acetylation correlate with decreased gene expression, histone hypo-acetylation may be a late event, as no hypo-acetylation is observed in 4-week-old R6/2 mice. Nevertheless, treatment with HDAC inhibitors corrects mRNA abnormalities through modification of histone proteins and may prove to be of therapeutic value in HD. PMID- 17409195 TI - This article was published online in error and has been removed by the publisher. PMID- 17409196 TI - Restriction landmark genome scanning identifies culture-induced DNA methylation instability in the human embryonic stem cell epigenome. AB - Widespread provision of human embryonic stem cells (hESCs) for therapeutic use, drug screening and disease modelling will require cell lines sustainable over long periods in culture. Since the short-term, in vitro culture of mammalian embryos can result in DNA methylation changes, the epigenetic stability of hESCs warrants investigation. Existing hESC lines have been derived and cultured under diverse conditions, providing the potential for programming differential changes into the epigenome that may result in inter-line variability over and above that inherited from the embryo. By examining the DNA methylation profiles of > 2000 genomic loci by Restriction Landmark Genome Scanning, we identified substantial inter-line epigenetic distance between six independently derived hESC lines. Lines were found to inherit further epigenetic changes over time in culture, with most changes arising in the earliest stages post-derivation. The loci affected varied between lines. The majority of culture-induced changes (82.3-87.5%) were stably inherited both within the undifferentiated cells and post-differentiation. Adapting a line to a serum-free culture system resulted in additional epigenetic instability. Overall 80.5% of the unstable loci uncovered in hESCs have been associated previously with an adult tumour phenotype. Our study shows that current methods of hESC propagation can rapidly programme stable and unpredictable epigenetic changes in the stem cell genome. This highlights the need for (i) novel screening strategies to determine the experimental utility and biosafety of hESCs and (ii) optimization and standardization of procedures for the derivation and culture of hESC lines that minimize culture-induced instability. PMID- 17409197 TI - Characterization and investigation of single nucleotide polymorphisms and a novel TLR2 mutation in the human TLR2 gene. AB - In the innate immune system, TLR2 plays a central role for the response to a wide variety of microbial and endogenous danger signals. A considerable number of genetic polymorphisms within the human TLR2 gene have been reported in non-coding and coding sequences. Except for the Arg753Gln variant, however, their clinical relevance is unclear and the assessment of the effects of amino acid substitutions on receptor function is lacking. In the present study, we have characterized all known single nucleotide polymorphisms (SNPs) of TLR2 for their functional relevance in transiently transfected HEK293 cells subsequently exposed to a specific stimulus. Among the known non-synonymous SNPs in the TLR2 coding sequence, four SNPs (Thr411Ile, Tyr715stop, Tyr715Lys and Arg753Gln) were found to be functionally relevant in our experimental setting. In addition, we identified a new mutation Arg447stop leading to a premature stop codon in the extracellular portion of the receptor. TLR2-specific stimulation of whole blood from two heterozygote donors of this mutation resulted in a reduced secretion of pro-inflammatory cytokines. Finally, we tested the prevalence of these functional genetic variants in 169 healthy individuals of Caucasian origin for the mutations in the extracellular domain and 106 individuals for the mutations in the intracellular domain of the receptor. Except for 10 heterozygote donors of the Arg753Gln variant determined to be prevalent in 9.4% of the tested individuals, none of the other SNPs was found in this population. PMID- 17409198 TI - Identification of polymorphic antioxidant response elements in the human genome. AB - Single nucleotide polymorphisms (SNPs) in transcription factor binding sites (TFBSs) may affect the binding of transcription factors, lead to differences in gene expression and phenotypes and therefore affect susceptibility to environmental exposure. We developed an integrated computational system for discovering functional SNPs in TFBSs in the human genome and predicting their impact on the expression of target genes. In this system, we (i) construct a position weight matrix (PWM) from a collection of experimentally discovered TFBSs; (ii) predict TFBSs in SNP sequences using the PWM and map SNPs to the upstream regions of genes; (iii) examine the evolutionary conservation of putative TFBSs by phylogenetic footprinting; (iv) prioritize candidate SNPs based on microarray expression profiles from tissues in which the transcription factor of interest is either deleted or over-expressed and (v) finally, analyze association of SNP genotypes with gene expression phenotypes. The application of our system has been tested to identify functional polymorphisms in the antioxidant response element (ARE), a cis-acting enhancer sequence found in the promoter region of many genes that encode antioxidant and Phase II detoxification enzymes/proteins. In response to oxidative stress, the transcription factor NRF2 (nuclear factor erythroid-derived 2-like 2) binds to AREs, mediating transcriptional activation of its responsive genes and modulating in vivo defense mechanisms against oxidative damage. Using our novel computational tools, we have identified a set of polymorphic AREs with functional evidence, showing the utility of our system to direct further experimental validation of genomic sequence variations that could be useful for identifying high-risk individuals. PMID- 17409199 TI - SOX9cre1, a cis-acting regulatory element located 1.1 Mb upstream of SOX9, mediates its enhancement through the SHH pathway. AB - SOX9 is a temporal and tissue-specific transcription factor involved in male sexual development and bone formation. Haploinsufficiency of SOX9 is known to cause campomelic dysplasia (CD). CD cases without SOX9 coding region mutations have been described in association with translocations that have breakpoints mapping as far as 932 kb upstream from the gene. These rearrangements suggest that position effects acting from a great distance regulate SOX9 gene expression. Studies of one such case (900 kb upstream to SOX9) have led to the delineation of a potential 2.1 kb cis-acting regulatory element 1.1 Mb upstream of SOX9, termed SOX9cre1. We investigated the role of this putative regulator in SOX9 expression. SOX9cre1 increases the activity of a minimal SOX9 promoter in reporter constructs in a dose-dependent and tissue-specific manner, consistent with an enhancer role. In silico studies identify a putative binding site within SOX9cre1 for GLI1, a downstream mediator of sonic hedgehog (SHH). Furthermore, the stimulation of primary human chondrocyte cells in culture with SHH increases endogenous SOX9 expression 3-fold. Electrophoresis mobility shift assay (EMSA) studies that demonstrate physical interactions between the GLI1 transcription factor and a putative binding site within SOX9cre1, as well as experiments in which reporter constructs are co-transfected with GLI1, suggest a direct interaction between GLI1 and SOX9cre1. GLI1-SOX9cre1 interactions are verified in chromatin immunoprecipitation experiments. These data support a direct molecular link between the Hh signaling pathway and SOX9 regulation, wherein SHH stimulates SOX9 through its mediator GLI1, and are consistent with a mechanism of SOX9 regulation through distal chromatin interactions. PMID- 17409200 TI - Triplet repeat mutation length gains correlate with cell-type specific vulnerability in Huntington disease brain. AB - Huntington disease is caused by the expansion of a CAG repeat encoding an extended glutamine tract in a protein called huntingtin. Here, we provide evidence supporting the hypothesis that somatic increases of mutation length play a role in the progressive nature and cell-selective aspects of HD pathogenesis. Results from micro-dissected tissue and individual laser-dissected cells obtained from human HD cases and knock-in HD mice indicate that the CAG repeat is unstable in all cell types tested although neurons tend to have longer mutation length gains than glia. Mutation length gains occur early in the disease process and continue to accumulate as the disease progresses. In keeping with observed patterns of cell loss, neuronal mutation length gains tend to be more prominent in the striatum than in the cortex of low-grade human HD cases, less so in more advanced cases. Interestingly, neuronal sub-populations of HD mice appear to have different propensities for mutation length gains; in particular, smaller mutation length gains occur in nitric oxide synthase-positive striatal interneurons (a relatively spared cell type in HD) compared with the pan-striatal neuronal population. More generally, the data demonstrate that neuronal changes in HD repeat length can be at least as great, if not greater, than those observed in the germline. The fact that significant CAG repeat length gains occur in non replicating cells also argues that processes such as inappropriate mismatch repair rather than DNA replication are involved in generating mutation instability in HD brain tissue. PMID- 17409201 TI - Evaluation of a commercial real-time PCR kit for detection of dengue virus in samples collected during an outbreak in Goiania, Central Brazil, in 2005. AB - In the past 2 decades, dengue has reemerged in Brazil as a significant public health problem. Clinicians demand a diagnostic test with high sensitivity that is applicable during the early symptomatic phase. We aimed to test two distinct molecular methods on samples from suspected dengue cases during an outbreak in Central Brazil. Acute-phase serum specimens from 254 patients suspected of having dengue were collected during 2005 in the city of Goiania, Central Brazil. Samples were blindly evaluated by real-time and multiplex PCR in addition to routine immunoglobulin M serology and virus culture. Overall, acute dengue was confirmed by serology, multiplex PCR, or virus isolation for 80% of patients (203/254). Another four patients presented real-time PCR-positive results as the unique marker of dengue. Higher real-time PCR positivity levels and viral loads were observed in the early symptomatic phase of disease (< or =5 days) than after this period. Multiplex and real-time PCR assays presented a high kappa agreement (0.85). According to multiplex PCR, 60 samples harbored dengue virus type 3 (DEN 3), 4 samples harbored DEN-2, and 1 sample displayed a pattern compatible with a double infection with DEN-2 and -3. The dengue virus real-time kit was found to be practical and adjustable for high throughput, to display the best performance in the early symptomatic phase of dengue cases, and to be valuable for confirming dengue diagnosis in a timely manner. PMID- 17409202 TI - Influence of disk preparation on detection of metallo-beta-lactamase-producing isolates by the combined disk assay. AB - The combined disk assay has been used for detection of metallo-beta-lactamase producing isolates. We have observed that the size of inhibition zones produced by many beta-lactam/metallo-beta-lactamase inhibitor (IMBL) combinations may differ depending on the way that the combined disks were prepared. Among the 10 beta-lactam/IMBL combinations tested, only the imipenem/EDTA combination produced similar results. PMID- 17409204 TI - Identification of an emerging pathogen, Mycobacterium massiliense, by rpoB sequencing of clinical isolates collected in the United States. AB - Mycobacterium massiliense is a rapidly growing mycobacterium that is indistinguishable from Mycobacterium chelonae/M. abscessus by partial 16S rRNA gene sequencing. We sequenced rpoB, sodA, and hsp65 genes from isolates previously identified as being M. chelonae/M. abscessus and identified M. massiliense from isolates from two patients with invasive disease representing the first reported cases in the United States. PMID- 17409205 TI - Frequent occult infection with Cytomegalovirus in cardiac transplant recipients despite antiviral prophylaxis. AB - Despite antiviral prophylaxis, a high percentage (over 90%) of heart transplant patients experience active cytomegalovirus (CMV) infection, diagnosed by detection of viral DNA in peripheral blood polymorphonuclear leukocytes within the first few months posttransplantation. Viral DNA was detected in mononuclear cells prior to detection in granulocytes from CMV-seropositive recipients (R+) receiving a heart from a CMV-seropositive donor (D+). Based on assessment of systemic infection in leukocyte populations, both R+ subgroups (R+/D- and R+/D+) experienced a greater infection burden than the R-/D+ subgroup, which was aggressively treated because of a higher risk of acute CMV disease. Despite widespread systemic infection in all at-risk patient subgroups, CMV DNA was rarely (< 3% of patients) detected in transplanted heart biopsy specimens. The R+ patients more frequently exceeded the 75th percentile of the CMV DNA copy number distribution in leukocytes (110 copies/10(5) polymorphonuclear leukocytes) than the R-/D+ subgroup. Therefore, active systemic CMV infection involving leukocytes is common in heart transplant recipients receiving prophylaxis to reduce acute disease. Infection of the transplanted organ is rare, suggesting that chronic vascular disease attributed to CMV may be driven by the consequences of systemic infection. PMID- 17409206 TI - WITHDRAWN: Molecular Characterization of VP4 and VP7 Genes of Nontypeable Strains Identifies a Novel P[28] Genotype in Porcine Rotaviruses. AB - This article has been retracted. PMID- 17409203 TI - Loss of bacterial diversity during antibiotic treatment of intubated patients colonized with Pseudomonas aeruginosa. AB - Management of airway infections caused by Pseudomonas aeruginosa is a serious clinical challenge, but little is known about the microbial ecology of airway infections in intubated patients. We analyzed bacterial diversity in endotracheal aspirates obtained from intubated patients colonized by P. aeruginosa by using 16S rRNA clone libraries and microarrays (PhyloChip) to determine changes in bacterial community compositions during antibiotic treatment. Bacterial 16S rRNA genes were absent from aspirates obtained from patients briefly intubated for elective surgery but were detected by PCR in samples from all patients intubated for longer periods. Sequencing of 16S rRNA clone libraries demonstrated the presence of many orally, nasally, and gastrointestinally associated bacteria, including known pathogens, in the lungs of patients colonized with P. aeruginosa. PhyloChip analysis detected the same organisms and many additional bacterial groups present at low abundance that were not detected in clone libraries. For each patient, both culture-independent methods showed that bacterial diversity decreased following the administration of antibiotics, and communities became dominated by a pulmonary pathogen. P. aeruginosa became the dominant species in six of seven patients studied, despite treatment of five of these six with antibiotics to which it was sensitive in vitro. Our data demonstrate that the loss of bacterial diversity under antibiotic selection is highly associated with the development of pneumonia in ventilated patients colonized with P. aeruginosa. Interestingly, PhyloChip analysis demonstrated reciprocal changes in abundance between P. aeruginosa and the class Bacilli, suggesting that these groups may compete for a similar ecological niche and suggesting possible mechanisms through which the loss of microbial diversity may directly contribute to pathogen selection and persistence. PMID- 17409207 TI - Epidemiologic distribution of the arginine catabolic mobile element among selected methicillin-resistant and methicillin-susceptible Staphylococcus aureus isolates. AB - We tested 214 Staphylococcus aureus isolates for the arcA locus of the arginine catabolic mobile element (ACME). All USA300 SCCmec IVa isolates, but no isolates containing other SCCmec subtypes, were arcA positive. arcA was also detected in selected methicillin-susceptible USA300 and methicillin-resistant USA100 isolates. DNA sequence analysis confirmed the integration of ACME in orfX. PMID- 17409208 TI - To test or not to test: a cost minimization analysis of susceptibility testing for patients with documented Candida glabrata fungemias. AB - This cost minimization analysis investigated the financial impact of the treatment of fungemias due to Candida glabrata from a hospital perspective using three competing alternatives: (i) performing in-house susceptibility testing on all C. glabrata isolates and changing patients to less expensive fluconazole therapy for isolates that test susceptible; (ii) susceptibility testing at outside laboratories with delayed deescalation to fluconazole if isolates test susceptible; and (iii) no routine susceptibility testing with full echinocandin treatment course. Sensitivity analyses and Monte Carlo simulation enhanced the robustness of the model through variation of all assumptions and costs. In the base case, the use of in-house testing displayed a cost advantage over the options of send-out testing and no susceptibility testing ($2,226 versus $2,410 versus $3,136, respectively). Sensitivity analyses determined that the cost of echinocandin therapy and the turnaround time for send-out testing had the potential to impact the base case model. The decision model indicated that in house susceptibility testing of C. glabrata isolates should result in lower overall treatment costs in patients with documented C. glabrata fungemias. PMID- 17409209 TI - Agreement between self- and clinician-collected specimen results for detection and typing of high-risk human papillomavirus in specimens from women in Gugulethu, South Africa. AB - We assessed the agreement in detection of high-risk human papillomavirus (HPV), as well as specific HPV types, between self- and clinician-obtained specimens for 450 women over 18 years of age attending a community health center in Gugulethu, South Africa. Both self-collected swabs and tampons had high agreement with clinician-obtained brushes when the Roche Reverse Line Blot Assay (RLBA) was used (for swabs, 86% concordance, with a kappa statistic [kappa] of 0.71; for tampons, 89% concordance, with kappa of 0.75). Agreement was lower, although still fair, with the Digene Hybrid Capture 2 test (HC2), with kappa higher for swabs than for tampons (for swabs, 81% concordance, with kappa of 0.61; for tampons, 82% concordance, with kappa of 0.55). Low-risk HPV types were nearly two times more common in self-collected specimens than in clinician-collected specimens tested by RLBA. All 15 women diagnosed with high-grade lesions by cytology tested positive for high-risk HPV with clinician-collected specimens tested by RLBA and HC2, while 11 out of 15 tested positive with self-collected specimens by HC2 and 5 out of 6 tested positive by RLBA. Self-collected specimens can provide valid specimens for HPV testing using nucleic acid amplification tests, although a few cytological abnormalities may be missed. PMID- 17409210 TI - Evaluation of real-time PCR laboratory-developed tests using analyte-specific reagents for cytomegalovirus quantification. AB - Viral load testing for cytomegalovirus (CMV) has become the standard for the diagnosis of infection and monitoring of therapy at many transplant centers. However, no viral load test has been approved by the FDA. Therefore, many laboratories rely on laboratory-developed assays. This study evaluated the performance characteristics of two real-time PCR tests developed using the artus CMV analyte-specific reagents (ASRs). One version is distributed by Abbott Molecular and the other by QIAGEN. For plasma specimens, the Abbott test had a limit of detection of 2.3 log10 copies/ml and a linear range up to at least 6.0 log10 copies/ml. Comparison of plasma viral loads using the Abbott test and the Roche Amplicor Monitor test showed a mean difference of -0.012 log10 copies/ml. In addition, the Abbott test viral loads correlated with the Digene Hybrid Capture assay ratios. Viral loads obtained from plasma specimens tested by the Abbott and QIAGEN tests were in very close agreement (mean difference, 0.144 log10 copies/ml). When the QIAGEN test was evaluated with the QIAGEN, MagNA Pure, and easyMAG extraction methods, the viral loads for all three methods were within 0.370 log10 copies/ml. Thus, there is good agreement between viral loads obtained by the different tests using the same extraction method or by the same test using different extraction methods. The availability of real-time PCR ASRs provides additional reagents that can be used for CMV viral load testing. PMID- 17409211 TI - Differences in clinical and molecular characteristics of skin and soft tissue methicillin-resistant Staphylococcus aureus isolates between two hospitals in Northern California. AB - Community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) skin and soft tissue infections (SSTI) are associated with SCCmec IV and Panton-Valentine leukocidin (PVL) genes. CO-MRSA epidemiologic studies suggest that genotypic variation exists within one geographic region. We compared MRSA genotypes and demographic and clinical characteristics of patients with CO-MRSA SSTI between two regional medical centers. We also examined factors associated with SCCmec IV and PVL carriage. A total of 279 MRSA SSTI isolates from 2000 to 2002 at San Francisco General Hospital (SFGH) and Stanford University Hospital (SUH) were genotyped by pulsed-field gel electrophoresis and PCR for SCCmec and PVL genes. Medical records were reviewed for clinical characteristics. Ninety-three percent and 69% of MRSA SSTI were caused by CO-MRSA at SFGH and SUH, respectively. Patients with CO-MRSA SSTI at SFGH were more likely to be nonwhite, younger, homeless, and have no previous exposure to health care (P < 0.01). SFGH CO-MRSA strains were more likely to carry SCCmec type IV and PVL genes (90% and 55%, respectively) than SUH strains (29% and 16%, respectively). In multivariate analyses, nonwhite ethnicity was associated with both SCCmec type IV and PVL carriage (odds ratio [OR] of 2.65 and 95% confidence interval [CI] of 1.19 to 5.95 and OR of 1.94 and 95% CI of 1.03 to 3.65, respectively). ST8:USA300:IV became the dominant clone at SFGH, but not at SUH, by 2002. Despite geographic proximity, CO-MRSA SSTI exhibited differing SCCmec types, PVL carriage, and clonal dynamics. CO-MRSA SSTI at SUH were more likely to represent feral isolates of nosocomial origin. Clinicians should assess for nosocomial and community risk factors, realizing that different populations with CO-MRSA SSTI may require separate antimicrobial strategies. PMID- 17409212 TI - Demonstration of presence of acanthamoeba mitochondrial DNA in brain tissue and cerebrospinal fluid by PCR in samples from a patient who died of granulomatous amebic encephalitis. PMID- 17409213 TI - Nosocomial outbreak of Corynebacterium striatum infection in patients with chronic obstructive pulmonary disease. AB - We describe an unusual cluster of Corynebacterium striatum infections in 21 patients with chronic obstructive pulmonary disease (COPD) admitted to a medium size respiratory unit. Eleven isolates from eight patients occurred simultaneously within a month. C. striatum is a potentially pathogenic microorganism with the ability to produce nosocomial infectious outbreaks and respiratory colonization in patients with advanced COPD. PMID- 17409214 TI - Serotype distribution and antimicrobial susceptibilities of nasopharyngeal isolates of Streptococcus pneumoniae from children hospitalized for acute respiratory illnesses in Hong Kong. AB - Five hundred nineteen Streptococcus pneumoniae isolates from nasopharyngeal aspirates of 3,157 children (age, <16 years) participating in a respiratory surveillance study in Hong Kong in 2005 and 2006 indicated that 64.9% and 37.2% of the isolates were not susceptible to penicillin and cefotaxime, respectively. The rate of potential coverage by the seven-valent conjugate vaccine was 72.3%, and the rate increased to 74.6% for serogroup-specific types. PMID- 17409215 TI - Addition of neuA, the gene encoding N-acylneuraminate cytidylyl transferase, increases the discriminatory ability of the consensus sequence-based scheme for typing Legionella pneumophila serogroup 1 strains. AB - The standard sequence-based method for the typing of Legionella pneumophila serogroup 1 strains was extended by using the gspA and neuA alleles. The use of neuA as a seventh allele for typing significantly increased the index of discrimination calculated for a panel of unrelated strains (from 0.932 to 0.963) and subdivided some known large common complexes (e.g., 1,4,3,1,1,1). This modification to the standard method is proposed as the method of choice in the epidemiological investigation of L. pneumophila infections. PMID- 17409216 TI - Multicenter evaluation of the new Abbott RealTime assays for quantitative detection of human immunodeficiency virus type 1 and hepatitis C virus RNA. AB - The analytical performances of the new Abbott RealTime hepatitis C virus (HCV) and human immunodeficiency virus type 1 viral load assays were compared at nine laboratories with different competitor assays. These included the Abbott LcX, Bayer Versant bDNA, Roche COBAS Amplicor, and Roche COBAS TaqMan assays. Two different protocols used during the testing period with and without a pre-m1000 RNA isolation spin were compared. The difference proved to be nonsignificant. A uracil-N-glycosylase (UNG) contamination control option in the HCV test for previous Roche COBAS Amplicor users was evaluated. It proved to decrease amplicon carryover by 100-fold independent of the amplicon input concentration. The protocol including UNG proved to overcome problems with false-positive negative controls. Comparison with other assays revealed only minor differences. The largest difference was observed between the Abbott HCV RealTime assay and the Roche COBAS Amplicor HCV Monitor version 2.0 assay. PMID- 17409217 TI - Metallo-beta-lactamase or extended-spectrum beta-lactamase: a wolf in sheep's clothing. AB - Diagnostic algorithms in commonly used automated bacterial identification systems fail to reliably identify a metallo-beta-lactamase in the Enterobacteriaceae. Misidentification as an extended-spectrum beta-lactamase may result in inappropriate dismissal of drugs such as aztreonam in favor of carbapenems, which may in turn select for a highly carabapenem resistant phenotype. PMID- 17409218 TI - Nuclear factor-kappaB accounts for the repressor effects of high stromal cell derived factor-1alpha levels on Tac1 expression in nontumorigenic breast cells. AB - Stromal cell-derived factor-1alpha (SDF-1alpha) is a CXC chemokine that interacts with CXCR4 receptor. Tac1 encodes peptides belonging to the tachykinins, including substance P. SDF-1alpha production is decreased in Tac1 knockdown breast cancer cells and is also reduced in these cancer cells following contact with bone marrow stroma when Tac1 expression is increased. Here, we report on the effects of relatively high and low SDF-1alpha levels on Tac1 expression in nontumorigenic breast cells MCF12A. Reporter gene assays, Northern analyses, and ELISA for substance P showed increased Tac1 expression at 20 and 50 ng/mL SDF 1alpha and reduced expression at 100 ng/mL. Omission of the untranslated region showed a dose-dependent effect of SDF-1alpha on reporter gene activity, suggesting that receptor desensitization cannot account for the suppressive effects at 100 ng/mL SDF-1alpha. Tac1 expression at high SDF-1alpha involves an intracellular signaling pathway that incorporates the activation of phosphatidylinositol 3-kinase-phosphoinositide-dependent kinase-1-AKT-nuclear factor-kappaB (NF-kappaB). The major repressive effect occurs via NF-kappaB located within exon 1. In summary, NF-kappaB is involved in the repression of Tac1 at higher levels of SDF-1alpha in MCF12A. These results are relevant to dysfunction of Tac1 in breast cancer cells and also provide insights on the behavior of breast cancer cells as they traverse across gradient changes of SDF 1alpha. PMID- 17409219 TI - Immune responses induced by replication-defective adenovirus expressing the C terminal portion of the Mycoplasma hyopneumoniae P97 adhesin. AB - Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, colonizes the respiratory cilia of affected swine, causing significant economic losses to swine production worldwide. Vaccination is the most cost-effective strategy for the control and prevention of this disease. The goal of this study was to design and evaluate a replication-defective recombinant adenovirus, rAdP97c, expressing the C-terminal portion of P97 adhesin (P97c), an important pathogenesis-associated protein of M. hyopneumoniae, as a new vaccine candidate against M. hyopneumoniae infection. P97c-specific immune responses were evaluated in BALB/c mice following intranasal and intramuscular inoculation with rAdP97c. Mice inoculated by both routes of immunization produced significant levels of specific immunoglobulin G (IgG) antibodies in the serum and in bronchoalveolar lavage fluids (BALs). Animals immunized intranasally also produced a significant level of P97c-specific IgA in BALs. Intramuscular inoculation of rAdP97c induced a systemic and mucosal Th1-type biased response, evidenced by the predominance of IgG2a in the serum and BALs, whereas intranasal inoculation resulted in a mixed Th1/Th2-type response (balanced levels of IgG1 and IgG2a) in both sytemic and mucosal compartments. P97c-specific antibodies were able to inhibit the growth of M. hyopneumoniae cells in vitro. These data suggest that rAdP97c vaccine may represent a new strategy for controlling infection by M. hyopneumoniae. PMID- 17409220 TI - Evaluation of the IMMY ALPHA Histoplasma antigen enzyme immunoassay for diagnosis of histoplasmosis marked by antigenuria. AB - The sensitivity of detection of antigenuria in patients positive by the MiraVista Diagnostics Histoplasma enzyme immunoassay (MVD EIA) was 44% with the IMMY ALPHA Histoplasma antigen EIA. The specificity was 84% with the IMMY EIA versus 98% with the MVD EIA. The correlation between assays for positive cases was weak (r(2) = 0.430). PMID- 17409221 TI - Rational peptide selection to detect human immunodeficiency virus type 1-specific T-cell responses under resource-limited conditions. AB - Understanding human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocyte responses is important for the development of vaccines and therapies. We describe a novel method for the rational selection of peptides that target stable regions of the HIV-1 genome, rich in epitopes specifically recognized by the study population. This method will be of particular use under resource/sample limited conditions. PMID- 17409222 TI - Experimental infection of neonatal foals with Rhodococcus equi triggers adult like gamma interferon induction. AB - Rhodococcus equi is a facultative intracellular pathogen that causes pneumonia in young foals but does not induce disease in immunocompetent adult horses. Clearance of R. equi depends mainly on gamma interferon (IFN-gamma) production by T lymphocytes, whereas the predominance of interleukin 4 (IL-4) is detrimental. Young foals, like neonates of many other species, are generally deficient in the ability to produce IFN-gamma. The objective of this study was to compare the cytokine profiles, as well as cell-mediated and antibody responses, of young foals to those of adult horses following intrabronchial challenge with R. equi. The lymphoproliferative responses of bronchial lymph node (BLN) cells to concanavalin A were significantly higher in foals than in adult horses. In contrast, adult horses had significantly higher lymphoproliferative responses to R. equi antigens than did foals. Infected foals had significantly lower IL-4 mRNA expression but significantly higher IFN-gamma expression and IFN-gamma/IL-4 ratio in R. equi-stimulated BLN lymphocytes than did infected adults. Infection with R. equi in foals resulted in a significant increase in the percentage of T lymphocytes and CD4(+) T lymphocytes in bronchoalveolar lavage fluid in association with a significant decrease in the percentage of these cell populations in BLNs. Infection of foals also resulted in a marked increase in serum immunoglobulin Ga (IgGa) and IgGb levels, resulting in concentrations in serum that were significantly higher than those of adult horses. This study demonstrates that the immune response to R. equi in foals is not biased toward IL 4 and is characterized by the predominant induction of IFN-gamma. PMID- 17409223 TI - Frequency, causes, and new challenges of indeterminate results in Western blot confirmatory testing for antibodies to human immunodeficiency virus. PMID- 17409224 TI - Global neuroinformatics: the International Neuroinformatics Coordinating Facility. PMID- 17409225 TI - Human peripersonal space: evidence from functional magnetic resonance imaging. PMID- 17409226 TI - Ketogenic diet metabolites reduce firing in central neurons by opening K(ATP) channels. AB - A low-carbohydrate ketogenic diet remains one of the most effective (but mysterious) treatments for severe pharmacoresistant epilepsy. We have tested for an acute effect of physiological ketone bodies on neuronal firing rates and excitability, to discover possible therapeutic mechanisms of the ketogenic diet. Physiological concentrations of ketone bodies (beta-hydroxybutyrate or acetoacetate) reduced the spontaneous firing rate of neurons in slices from rat or mouse substantia nigra pars reticulata. This region is thought to act as a "seizure gate," controlling seizure generalization. Consistent with an anticonvulsant role, the ketone body effect is larger for cells that fire more rapidly. The effect of ketone bodies was abolished by eliminating the metabolically sensitive K(ATP) channels pharmacologically or by gene knock-out. We propose that ketone bodies or glycolytic restriction treat epilepsy by augmenting a natural activity-limiting function served by K(ATP) channels in neurons. PMID- 17409227 TI - Removal of GABA within adult modulatory systems alters electrical coupling and allows expression of an embryonic-like network. AB - The maturation and operation of neural networks are known to depend on modulatory neurons. However, whether similar mechanisms may control both adult and developmental plasticity remains poorly investigated. To examine this issue, we have used the lobster stomatogastric nervous system (STNS) to investigate the ontogeny and role of GABAergic modulatory neurons projecting to small pattern generating networks. Using immunocytochemistry, we found that modulatory input neurons to the stomatogastric ganglion (STG) express GABA only after metamorphosis, a time that coincides with the developmental switch from a single to multiple pattern generating networks within the STNS. We demonstrate that blocking GABA synthesis with 3-mercapto-propionic acid within the adult modulatory neurons results in the reconfiguration of the distinct STG networks into a single network that generates a unified embryonic-like motor pattern. Using dye-coupling experiments, we also found that gap-junctional coupling is greater in embryos and GABA-deprived adults exhibiting the unified motor pattern compared with control adults. Furthermore, GABA was found to diminish directly the extent and strength of electrical coupling within adult STG networks. Together, these observations suggest the acquisition of a GABAergic phenotype by modulatory neurons after metamorphosis may induce the reconfiguration of the single embryonic network into multiple adult networks by directly decreasing electrical coupling. The findings also suggest that adult neural networks retain the ability to express typical embryonic characteristics, indicating that network ontogeny can be reversed and that changes in electrical coupling during development may allow the segregation of multiple distinct functional networks from a single large embryonic network. PMID- 17409228 TI - Beta-site amyloid precursor protein cleaving enzyme 1 levels become elevated in neurons around amyloid plaques: implications for Alzheimer's disease pathogenesis. AB - Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1) (beta-secretase) initiates generation of beta-amyloid (Abeta), which plays an early role in Alzheimer's disease (AD). BACE1 levels are increased in postmortem AD brain, suggesting BACE1 elevation promotes Abeta production and AD. Alternatively, the BACE1 increase may be an epiphenomenon of late-stage AD. To distinguish between these possibilities, we analyzed BACE1 elevation using a highly specific BACE1 antibody, BACE-Cat1, made in BACE1-/- mice, which mount a robust anti-BACE1 immune response. Previous BACE1 immunohistochemical studies lack consistent results because typical BACE1 antibodies produce nonspecific background, but BACE Cat1 immunolabels BACE1 only. BACE1 elevation was recapitulated in two amyloid precursor protein (APP) transgenic mouse lines. 5XFAD mice form amyloid plaques at young ages and exhibit neuron loss. In contrast, Tg2576 form plaques at a more advanced age and do not show cell death. These two mouse lines allow differentiation between early Abeta-induced events and late phenomena related to neuron death. BACE1 levels became elevated in parallel with amyloid burden in each APP transgenic, starting early in 5XFAD and late in Tg2576. The increase in BACE1 protein occurred without any change in BACE1 mRNA level, indicating a posttranscriptional mechanism. In APP transgenic and AD brains, high BACE1 levels were observed in an annulus around Abeta42-positive plaque cores and colocalized with neuronal proteins. These results demonstrate that amyloid plaques induce BACE1 in surrounding neurons at early stages of pathology before neuron death occurs. We conclude that BACE1 elevation is most likely triggered by the amyloid pathway and may drive a positive-feedback loop in AD. PMID- 17409229 TI - Accumulation of pathological tau species and memory loss in a conditional model of tauopathy. AB - Neurofibrillary tangles (NFTs) are a pathological hallmark of Alzheimer's disease and other tauopathies, but recent studies in a conditional mouse model of tauopathy (rTg4510) have suggested that NFT formation can be dissociated from memory loss and neurodegeneration. This suggests that NFTs are not the major neurotoxic tau species, at least during the early stages of pathogenesis. To identify other neurotoxic tau protein species, we performed biochemical analyses on brain tissues from the rTg4510 mouse model and then correlated the levels of these tau proteins with memory loss. We describe the identification and characterization of two forms of tau multimers (140 and 170 kDa), whose molecular weight suggests an oligomeric aggregate, that accumulate early in the pathogenic cascade in this mouse model. Similar tau multimers were detected in a second mouse model of tauopathy (JNPL3) and in tissue from patients with Alzheimer's disease and FTDP-17 (frontotemporal dementia and parkinsonism linked to chromosome 17). Moreover, levels of the tau multimers correlated consistently with memory loss at various ages in the rTg4510 mouse model. Our findings suggest that accumulation of early-stage aggregated tau species, before the formation of NFT, is associated with the development of functional deficits during the pathogenic progression of tauopathy. PMID- 17409230 TI - Subcellular arrangement of molecules for 2-arachidonoyl-glycerol-mediated retrograde signaling and its physiological contribution to synaptic modulation in the striatum. AB - Endogenous cannabinoids (endocannabinoids) mediate retrograde signals for short- and long-term suppression of transmitter release at synapses of striatal medium spiny (MS) neurons. An endocannabinoid, 2-arachidonoyl-glycerol (2-AG), is synthesized from diacylglycerol (DAG) after membrane depolarization and Gq coupled receptor activation. To understand 2-AG-mediated retrograde signaling in the striatum, we determined precise subcellular distributions of the synthetic enzyme of 2-AG, DAG lipase-alpha (DAGLalpha), and its upstream metabotropic glutamate receptor 5 (mGluR5) and muscarinic acetylcholine receptor 1 (M1). DAGLalpha, mGluR5, and M1 were all richly distributed on the somatodendritic surface of MS neurons, but their subcellular distributions were different. Although mGluR5 and DAGLalpha levels were highest in spines and accumulated in the perisynaptic region, M1 level was lowest in spines and was rather excluded from the mGluR5-rich perisynaptic region. These subcellular arrangements suggest that mGluR5 and M1 might differentially affect endocannabinoid-mediated, depolarization-induced suppression of inhibition (DSI) and depolarization-induced suppression of excitation (DSE) in MS neurons. Indeed, mGluR5 activation enhanced both DSI and DSE, whereas M1 activation enhanced DSI only. Importantly, DSI, DSE, and receptor-driven endocannabinoid-mediated suppression were all abolished by the DAG lipase inhibitor tetrahydrolipstatin, indicating 2-AG as the major endocannabinoid mediating retrograde suppression at excitatory and inhibitory synapses of MS neurons. Accordingly, CB1 cannabinoid receptor, the main target of 2-AG, was present at high levels on GABAergic axon terminals of MS neurons and parvalbumin-positive interneurons and at low levels on excitatory corticostriatal afferents. Thus, endocannabinoid signaling molecules are arranged to modulate the excitability of the MS neuron effectively depending on cortical activity and cholinergic tone as measured by mGluR5 and M1 receptors, respectively. PMID- 17409231 TI - Mice lacking protease nexin-1 show delayed structural and functional recovery after sciatic nerve crush. AB - Multiple molecular mechanisms influence nerve regeneration. Because serine proteases were shown to affect peripheral nerve regeneration, we performed nerve crush experiments to study synapse reinnervation in adult mice lacking the serpin protease nexin-1 (PN-1). PN-1 is a potent endogenous inhibitor of thrombin, trypsin, tissue plasminogen activators (tPAs), and urokinase plasminogen activators. Compared with the wild type, a significant delay in synapse reinnervation was detected in PN-1 knock-out (KO) animals, which was associated with both reduced proliferation and increased apoptosis of Schwann cells. Various factors known to affect Schwann cells were also altered. Fibrin deposits, tPA activity, mature BDNF, and the low-affinity p75 neurotrophin receptor were increased in injured sciatic nerves of mutant mice. To test whether the absence of PN-1 in Schwann cells or in the axon caused delay in reinnervation, PN-1 was overexpressed exclusively in the nerves of PN-1 KO mice. Neuronal PN-1 expression did not rescue the delayed reinnervation. The results suggest that Schwann cell derived PN-1 is crucial for proper reinnervation through its contribution to the autocrine control of proliferation and survival. Thus, the precise balance between distinct proteases and serpins such as PN-1 can modulate the overall impact on the kinetics of recovery. PMID- 17409232 TI - Prenatal exposure to elevated NT3 disrupts synaptic selectivity in the spinal cord. AB - Monosynaptic connections between muscle spindle (Ia) afferents and motoneurons (MNs), the central portion of the stretch reflex circuit, are highly specific, but the mechanisms underlying this specificity are primarily unknown. In this study, we report that embryonic overexpression of neurotrophin-3 (NT3) in muscles disrupts the development of these specific Ia-MN connections, using transgenic (mlc/NT3) mice that express elevated levels of NT3 in muscles during development. In mlc/NT3 mice, there is a substantial increase in the amplitudes of monosynaptic EPSPs evoked by Ia afferents in MNs as measured with extracellular recordings from ventral roots. Despite this increased functional projection of Ia afferents, there is no obvious change in the anatomical density of Ia projections into the ventral horn of the spinal cord. Intracellular recordings from MNs revealed a major disruption in the pattern of Ia-MN connections. In addition to the normal connections between Ia afferents and MNs supplying the same muscle, there were also strong monosynaptic inputs from Ia afferents supplying unrelated muscles, which explains the increase seen in extracellular recordings. There was also a large variability in the strength of Ia input to individual MNs, both from correct and incorrect Ia afferents. Postnatal muscular administration of NT3 did not cause these changes in connectivity. These results indicate that prenatal exposure to elevated levels of NT3 disrupts the normal mechanisms responsible for synaptic selectivity in the stretch reflex circuit. PMID- 17409233 TI - Specific alterations of extracellular endocannabinoid levels in the nucleus accumbens by ethanol, heroin, and cocaine self-administration. AB - Ethanol and opiate self-administration are sensitive to manipulations of cannabinoid CB1 receptor function and, from this, a role for the endogenous cannabinoid system in the modulation of drug reward has been hypothesized. However, direct in vivo evidence of drug-induced alterations in brain endocannabinoid (eCB) formation has been lacking. To address this issue, we explored the effect of drug self-administration on interstitial eCB levels in the nucleus accumbens (NAc) shell using in vivo microdialysis. Ethanol, heroin, and cocaine were compared because the rewarding properties of ethanol and heroin are reduced by CB1 receptor inactivation, whereas cocaine reward is less sensitive to these manipulations. Ethanol self-administration significantly increased dialysate 2-arachidonoylglycerol (2-AG) levels with no concomitant change in dialysate anandamide (AEA) concentrations. Conversely, heroin self-administration significantly increased dialysate AEA levels, and induced a subtle but significant decrease in dialysate 2-AG levels. In each case, the relative change in dialysate eCB content was significantly correlated with the amount of drug consumed. In contrast, cocaine self-administration did not alter dialysate levels of either AEA or 2-AG. Local infusion of the CB1 antagonist SR 141716A into the NAc significantly reduced ethanol, but not cocaine, self-administration. Together with our previous observation that intra-NAc SR 141716A reduces heroin self administration, these data provide novel in vivo support for an eCB involvement in the motivational properties of ethanol and heroin but not cocaine. Furthermore, the selective effects of ethanol and heroin on interstitial 2-AG and AEA provide new insight into the distinct neurochemical profiles produced by these two abused substances. PMID- 17409234 TI - HIV-1 Vpr causes neuronal apoptosis and in vivo neurodegeneration. AB - Despite the introduction of highly active antiretroviral therapy, dementia caused by human immunodeficiency virus-1 (HIV-1) infection remains a devastating and common neurological disorder. Although the mechanisms governing neurodegeneration during HIV-1 infection remain uncertain, the HIV-1 accessory protein, viral protein R (Vpr), has been proposed as a neurotoxic protein. Herein, we report that Vpr protein and transcript were present in the brains of HIV-infected persons. Moreover, soluble Vpr caused neuronal apoptosis, involving cytochrome c extravasation, p53 induction, and activation of caspase-9 while exerting a depressive effect on whole-cell currents in neurons (p < 0.05), which was inhibited by iberiotoxin. Vpr-activated glial cells secreted neurotoxins in a concentration-dependent manner (p < 0.001). Transgenic (Tg) mice expressing Vpr in brain monocytoid cells displayed the transgene principally in the basal ganglia (p < 0.05) and cerebral cortex (p < 0.01) compared with hindbrain expression. Vpr was released from cultured transgenic macrophages, which was cytotoxic to neurons and was blocked by anti-Vpr antibody (p < 0.05). Neuronal injury was observed in Tg animals compared with wild-type littermates, chiefly affecting GAD65 (p < 0.01) and vesicular acetylcholine transferase (p < 0.001) immunopositive neuronal populations in the basal ganglia. There was also a loss of subcortical synaptophysin (p < 0.001) immunoreactivity as well as an increase in activated caspase-3, which was accompanied by a hyperexcitable neurobehavioral phenotype (p < 0.05). Thus, HIV-1 Vpr caused neuronal death through convergent pathogenic mechanisms with ensuing in vivo neurodegeneration, yielding new insights into the mechanisms by which HIV-1 injures the nervous system. PMID- 17409235 TI - Pyrrolidine dithiocarbamate activates Akt and improves spatial learning in APP/PS1 mice without affecting beta-amyloid burden. AB - Pyrrolidine dithiocarbamate (PDTC) is a clinically tolerated inhibitor of nuclear factor-kappaB (NF-kappaB), antioxidant and antiinflammatory agent, which provides protection in brain ischemia models. In neonatal hypoxia-ischemia model, PDTC activates Akt and reduces activation of glycogen synthase kinase 3beta (GSK 3beta). Because chronic inflammation, oxidative stress, and increased GSK-3beta activity are features of Alzheimer's disease (AD) pathology, we tested whether PDTC reduces brain pathology and improves cognitive function in a transgenic animal model of AD. A 7 month oral treatment with PDTC prevented the decline in cognition in AD mice without altering beta-amyloid burden or gliosis. Moreover, marked oxidative stress and activation of NF-kappaB were not part of the brain pathology. Instead, the phosphorylated form of GSK-3beta was decreased in the AD mouse brain, and PDTC treatment increased the phosphorylation of Akt and GSK 3beta. Also, PDTC treatment increased the copper concentration in the brain. In addition, PDTC rescued cultured hippocampal neurons from the toxicity of oligomeric Abeta and reduced tau phosphorylation in the hippocampus of AD mice. Finally, astrocytic glutamate transporter GLT-1, known to be regulated by Akt pathway, was decreased in the transgenic AD mice but upregulated back to the wild type levels by PDTC treatment. Thus, PDTC may improve spatial learning in AD by interfering with Akt-GSK pathway both in neurons and astrocytes. Because PDTC is capable of transferring external Cu2+ into a cell, and, in turn, Cu2+ is able to activate Akt, we hypothesize that PDTC provides the beneficial effect in transgenic AD mice through Cu2+-activated Akt pathway. PMID- 17409236 TI - Telomere protection mechanisms change during neurogenesis and neuronal maturation: newly generated neurons are hypersensitive to telomere and DNA damage. AB - Telomeres are DNA-protein complexes at the ends of eukaryotic chromosomes that play an important role in maintaining the integrity of the genome. In proliferative stem cells and cancer cells, telomere length is maintained by telomerase, and telomere structure and functions are regulated by telomere associated proteins. We find that telomerase levels are high in embryonic cortical neural progenitor cells (NPCs) and low in newly generated neurons (NGNs) and mature neurons (MNs). In contrast, telomere repeat-binding factor 2 (TRF2) expression is undetectable in early brain development in vivo and in cultured NPCs and is expressed at progressively higher levels as NPCs cease proliferation and differentiate into postmitotic neurons. The telomere-disrupting agent telomestatin induces a DNA damage response and apoptosis in NGNs (which have low levels of TRF2 and telomerase), whereas NPCs (which have high levels of telomerase) and MNs (which have high levels of TRF2) are resistant to telomere damage. Overexpression of TRF2 in NGNs protects them against death induced by telomestatin and other DNA-damaging agents. Knockdown of TRF2 expression in MNs and knock-out of telomerase reverse transcriptase in NPCs increased their sensitivity to telomere- and DNA-damaging agents but did not affect the vulnerability of NGNs. These findings suggest that TRF2 and telomerase function as distinct telomere protection mechanisms during the processes of neurogenesis and neuronal maturation and that hypersensitivity of NGNs to telomere damage results from relative deficiencies of both telomerase and TRF2. PMID- 17409237 TI - The doublecortin-expressing population in the developing and adult brain contains multipotential precursors in addition to neuronal-lineage cells. AB - Doublecortin (DCX) has recently been promulgated as a selective marker of cells committed to the neuronal lineage in both the developing and the adult brain. To explore the potential of DCX-positive (DCX+) cells more stringently, these cells were isolated by flow cytometry from the brains of transgenic mice expressing green fluorescent protein under the control of the DCX promoter in embryonic, early postnatal, and adult animals. It was found that virtually all of the cells (99.9%) expressing high levels of DCX (DCX(high)) in the embryonic brain coexpressed the neuronal marker betaIII-tubulin and that this population contained no stem-like cells as demonstrated by lack of neurosphere formation in vitro. However, the DCX+ population from the early postnatal brain and the adult subventricular zone and hippocampus, which expressed low levels of DCX (DCX(low)), was enriched for neurosphere-forming cells, with only a small subpopulation of these cells coexpressing the neuronal markers betaIII-tubulin or microtubule-associated protein 2. Similarly, the DCX(low) population from embryonic day 14 (E14) brain contained neurosphere-forming cells. Only the postnatal cerebellum and adult olfactory bulb contained some DCX(high) cells, which were shown to be similar to the E14 DCX(high) cells in that they had no stem cell activity. Electrophysiological studies confirmed the heterogeneous nature of DCX+ cells, with some cells displaying characteristics of immature or mature neurons, whereas others showed no neuronal characteristics whatsoever. These results indicate that DCX(high) cells, regardless of location, are restricted to the neuronal lineage or are bone fide neurons, whereas some DCX(low) cells retain their multipotentiality. PMID- 17409238 TI - Triangulating a cognitive control network using diffusion-weighted magnetic resonance imaging (MRI) and functional MRI. AB - The ability to stop motor responses depends critically on the right inferior frontal cortex (IFC) and also engages a midbrain region consistent with the subthalamic nucleus (STN). Here we used diffusion-weighted imaging (DWI) tractography to show that the IFC and the STN region are connected via a white matter tract, which could underlie a "hyperdirect" pathway for basal ganglia control. Using a novel method of "triangulation" analysis of tractography data, we also found that both the IFC and the STN region are connected with the presupplementary motor area (preSMA). We hypothesized that the preSMA could play a conflict detection/resolution role within a network between the preSMA, the IFC, and the STN region. A second experiment tested this idea with functional magnetic resonance imaging (fMRI) using a conditional stop-signal paradigm, enabling examination of behavioral and neural signatures of conflict-induced slowing. The preSMA, IFC, and STN region were significantly activated the greater the conflict-induced slowing. Activation corresponded strongly with spatial foci predicted by the DWI tract analysis, as well as with foci activated by complete response inhibition. The results illustrate how tractography can reveal connections that are verifiable with fMRI. The results also demonstrate a three way functional-anatomical network in the right hemisphere that could either brake or completely stop responses. PMID- 17409239 TI - Plasma membrane Ca2+ ATPase 2 contributes to short-term synapse plasticity at the parallel fiber to Purkinje neuron synapse. AB - Plasma membrane Ca2+ ATPase 2 (PMCA2) is a fast, highly effective mechanism to control resting cytosolic Ca2+ and Ca2+ excursions in neurons and other excitable cells. The strong expression of PMCA2 in the cerebellum and the cerebellar behavioral deficits presented by PMCA2-/- knock-out mice all point to its importance for cerebellar circuit dynamics. Here, we provide direct functional evidence for the influence of presynaptic PMCA2-mediated Ca2+ extrusion for short term plasticity at cerebellar parallel fiber to Purkinje neuron synapses. Dramatic structural alterations to the Purkinje neurons in the absence of PMCA2 also suggest a strong influence of this fast PMCA2 isoform for development and maintenance of cerebellar function. PMID- 17409240 TI - Microstimulation of the midbrain tegmentum creates learning signals for saccade adaptation. AB - Error signals are vital to motor learning. However, we know little about pathways that transmit error signals for learning in voluntary movements. Here we show that microstimulation of the midbrain tegmentum can induce learning in saccadic eye movements in monkeys. Weak electrical stimuli delivered approximately 200 ms after saccades in one horizontal direction produced gradual and marked changes in saccade gain. The spatial and temporal characteristics of the produced changes were similar to those of adaptation induced by real visual error. When stimulation was applied after saccades in two different directions, endpoints of these saccades gradually shifted in the same direction in two dimensions. We conclude that microstimulation created powerful learning signals that dictate the direction of adaptive shift in movement endpoints. Our findings suggest that the error signals for saccade adaptation are conveyed in a pathway that courses through the midbrain tegmentum. PMID- 17409241 TI - Altered NMDA receptor trafficking in a yeast artificial chromosome transgenic mouse model of Huntington's disease. AB - Overactivation of NMDA receptors (NMDARs) is believed to play a role in degeneration of striatal medium-sized spiny neurons (MSNs) in Huntington's disease (HD). This hereditary disorder is caused by an expansion >35 in the polyglutamine (polyQ) region of the protein huntingtin (htt). Previous work has shown that NMDAR current, calcium signaling, and/or toxicity are enhanced in striatal MSNs in a variety of transgenic mice and cellular models of HD, but whether the enhancement is specific for MSNs or correlated with mutant htt (mhtt) polyQ length is not known. Furthermore, the mechanism underlying the increase in NMDAR activity has not been elucidated. Here we report polyQ length-dependent enhancement of peak NMDAR current density by mhtt in cultured MSNs, but not cortical neurons, from the yeast artificial chromosome (YAC) transgenic HD mouse model. We also observed a shift of NMDAR subunits NR1 and NR2B from internal pools to the plasma membrane and a significantly faster rate of NMDAR insertion to the surface in YAC72 MSNs. In comparing YAC72 with wild-type striatal tissue, subcellular fractionation revealed a relative enrichment of NR1 C2'-containing NMDARs in the vesicle/microsome-enriched fraction, and coimmunoprecipitation experiments demonstrated an increased proportion of NR1 C2' isoforms associated with NR2 subunits, which may contribute to faster forward trafficking of these receptors. Our results suggest that altered NMDAR trafficking may underlie potentiation of NMDAR-mediated current and toxicity in the YAC72 HD mouse model. This polyQ length-dependent, neuronal-specific change in NMDAR activity induced by mhtt may contribute to selective neuronal degeneration in HD. PMID- 17409242 TI - Electrophysiological properties of AMPA receptors are differentially modulated depending on the associated member of the TARP family. AB - The family of AMPA receptors is encoded by four genes that are differentially spliced to result in the flip or flop versions of the four subunits GluR1 to GluR4. GluR2 is further modified at the so-called Q/R site by posttranscriptional RNA editing. Delivery of AMPA receptors to the plasma membrane and synaptic trafficking are controlled by transmembrane AMPA receptor regulatory proteins (TARPs). Additionally, TARPs influence essential electrophysiological properties of AMPA receptor channels such as desensitization and agonist efficacies. Here, we compare the influence of all known TARPs (gamma2, gamma3, gamma4, and gamma8) on agonist-induced currents of the four AMPA receptor subunits, including flip and flop splice variants and editing variants. We show that, although agonist induced currents of all homomeric AMPA receptor subunits as well as all heteromeric combinations tested are significantly potentiated when coexpressed with members of the TARP family in Xenopus laevis oocytes, the extent of TARP mediated increase in agonist-induced responses is highly dependent on both the AMPA receptor subunit and the coexpressed TARP. Moreover, we demonstrate that the splice variant of the AMPA receptor plays a key role in determining the modulation of electrophysiological properties by associated TARPs. We furthermore present evidence that individual TARP-AMPA receptor interactions control the degree of desensitization of AMPA receptors. Consequently, because of their subunit-specific impact on the electrophysiological properties, TARPs play a major role as modulatory subunits of AMPA receptors and thus contribute to the functional diversity of AMPA receptors encountered in the CNS. PMID- 17409243 TI - Selective retrieval of abstract semantic knowledge in left prefrontal cortex. AB - Research into the representation and processing of conceptual knowledge has typically associated perceptual facts with sensory brain regions and executive retrieval mechanisms with the left prefrontal cortex. However, this dichotomy between knowledge content and retrieval processes leaves unanswered how the brain supports concepts less reliant on direct sensory experiences. We used neuroimaging methods to investigate whether an increased abstractness in semantic decisions, in contrast to increased response difficulty, is associated with increased left prefrontal activation. Participants were presented with concrete animal names and asked to verify sensory and abstract properties that corresponded only to the animal category. Candidate semantic regions were localized in left inferior, frontopolar, and dorsolateral prefrontal cortex in contrast to a pseudoword control. Activity in each of these prefrontal regions was associated with significantly increased activity for abstract relative to sensory semantic decisions, regardless of increased response difficulty and even when controlling for the response times of participants. These results suggest that more abstract, or verbally-mediated, semantic knowledge of concrete items, in contrast to more sensory-based properties, is specifically supported by the left prefrontal cortex. Semantic retrieval mechanisms may rely on abstract representations, likely coded through a verbal format, to mediate task demands when perceptual information is insufficient. PMID- 17409244 TI - Broca's area supports enhanced visuospatial cognition in orchestral musicians. AB - We provide neurobehavioral evidence supporting the transferable benefit of music training to alter brain function and enhance cognitive performance in a nonmusical visuospatial task in professional orchestral musicians. In particular, orchestral musicians' performance on a three-dimensional mental rotation (3DMR) task exhibited the behavioral profile normally only attained after significant practice, supporting the suggestion that these musicians already possessed well developed neural circuits to support 3DMR. Furthermore, functional magnetic resonance imaging revealed that only orchestral musicians showed significantly increased activation in Broca's area, in addition to the well known visuospatial network, which was activated in both musicians and nonmusicians who were matched on age, sex, and verbal intelligence. We interpret these functional neuroimaging findings to reflect preferential recruitment of Broca's area, part of the neural substrate supporting sight reading and motor-sequence organization underpinning musical performance, to subserve 3DMR in musicians. Our data, therefore, provide convergent behavioral and neurofunctional evidence supporting the suggestion that development of the sight-reading skills of musical performance alters brain circuit organization which, in turn, confers a wider cognitive benefit, in particular, to nonmusical visuospatial cognition in professional orchestral musicians. PMID- 17409245 TI - Timing-dependent modulation of associative plasticity by general network excitability in the human motor cortex. AB - Associative neuroplasticity, which encompasses the modification of synaptic strength by coactivation of two synaptic inputs, has been linked to learning processes. Because unlimited plasticity destabilizes neuronal networks, homeostatic rules were proposed and experimentally proven that control for the amount and direction of plasticity dependent on background network activity. Accordingly, low background activity would enhance facilitatory plasticity, whereas high background activity would inhibit it. However, the impact of background excitability on associative plasticity has not been studied so far in humans. Facilitatory associative plasticity was induced by paired associative stimulation (PAS) in the human motor cortex, whereas background activity was enhanced or diminished by transcranial direct current stimulation (tDCS). When applied before PAS, excitability-enhancing tDCS also boosted the efficacy of PAS, whereas excitability-diminishing tDCS turned it into inhibition. Thus, previous background activity does not influence associative plasticity homeostatically. When tDCS and PAS were applied simultaneously, now in accordance with homeostatic rules of neuroplasticity, reduced background activity resulted in a prolonged excitability enhancement by PAS, whereas enhanced background activity turned it into inhibition. We conclude that background network activity can influence associative plasticity homeostatically. However, only simultaneous modulation of both parameters is in accordance with homeostatic concepts. These findings might be of importance for the development of plasticity-inducing stimulation protocols supporting information processing in humans. PMID- 17409246 TI - Dopamine receptor activation modulates GABA neuron migration from the basal forebrain to the cerebral cortex. AB - GABA neurons of the cerebral cortex and other telencephalic structures are produced in the basal forebrain and migrate to their final destinations during the embryonic period. The embryonic basal forebrain is enriched in dopamine and its receptors, creating a favorable environment for dopamine to influence GABA neuron migration. However, whether dopamine receptor activation can influence GABA neuron migration is not known. We show that dopamine D1 receptor activation promotes and D2 receptor activation decreases GABA neuron migration from the medial and caudal ganglionic eminences to the cerebral cortex in slice preparations of embryonic mouse forebrain. Slice preparations from D1 or D2 receptor knock-out mouse embryos confirm the findings. In addition, D1 receptor electroporation into cells of the basal forebrain and pharmacological activation of the receptor promote migration of the electroporated cells to the cerebral cortex. Analysis of GABA neuron numbers in the cerebral wall of the dopamine receptor knock-out mouse embryos further confirmed the effects of dopamine receptor activation on GABA neuron migration. Finally, dopamine receptor activation mobilizes striatal neuronal cytoskeleton in a manner consistent with the effects on neuronal migration. These data show that impairing the physiological balance between D1 and D2 receptors can alter GABA neuron migration from the basal forebrain to the cerebral cortex. The intimate relationship between dopamine and GABA neuron development revealed here may offer novel insights into developmental disorders such as schizophrenia, attention deficit or autism, and fetal cocaine exposure, all of which are associated with dopamine and GABA imbalance. PMID- 17409247 TI - T-type and L-type Ca2+ conductances define and encode the bimodal firing pattern of vestibulocerebellar unipolar brush cells. AB - Cerebellar unipolar brush cells (UBCs) are glutamatergic interneurons that receive direct input from vestibular afferents in the form of a unique excitatory synapse on their dendritic brush. UBCs constitute independent relay lines for vestibular signals, and their inherent properties most likely determine how vestibular activity is encoded by the cerebellar cortex. We now demonstrate that UBCs are bimodal cells; they can either fire high-frequency bursts of action potentials when stimulated from hyperpolarized potentials or discharge tonically during sustained depolarizations. The two functional states can be triggered by physiological-like activity of the excitatory input and are encoded by distinct Ca2+-signaling systems. By combining complementary strategies, consisting of molecular and electrophysiological analysis and of ultrafast acousto-optical deflector-based two-photon imaging, we unraveled the identity and the subcellular localization of the Ca2+ conductances activating in each mode. Fast inactivating T-type Ca2+ channels produce low-threshold spikes, which trigger the high frequency bursts and generate powerful Ca2+ transients in the brush and, to a much lesser extent, in the soma. The tonic firing mode is encoded by a signalization system principally composed of L-type channels. Ca2+ influx during tonic firing produces a linear representation of the spike rate of the cell in the form of a widespread and sustained Ca2+ concentration increase and regulates cellular excitability via BK potassium channels. The bimodal firing pattern of UBCs may underlie different coding strategies of the vestibular input by the cerebellum, thus likely increasing the computational power of this structure. PMID- 17409248 TI - Brain oscillations control timing of single-neuron activity in humans. AB - A growing body of animal research suggests that neurons represent information not only in terms of their firing rates but also by varying the timing of spikes relative to neuronal oscillations. Although researchers have argued that this temporal coding is critical in human memory and perception, no supporting data from humans have been reported. This study provides the first analysis of the temporal relationship between brain oscillations and single-neuron activity in humans. Recording from 1924 neurons, we find that neuronal activity in various brain regions increases at specific phases of brain oscillations. Neurons in widespread brain regions were phase locked to oscillations in the theta- (4-8 Hz) and gamma- (30-90 Hz) frequency bands. In hippocampus, phase locking was prevalent in the delta- (1-4 Hz) and gamma-frequency bands. Individual neurons were phase locked to various phases of theta and delta oscillations, but they only were active at the trough of gamma oscillations. These findings provide support for the temporal-coding hypothesis in humans. Specifically, they indicate that theta and delta oscillations facilitate phase coding and that gamma oscillations help to decode combinations of simultaneously active neurons. PMID- 17409250 TI - Molecular nature of anomalous L-type calcium channels in mouse cerebellar granule cells. AB - Single-channel analysis revealed the existence of neuronal L-type Ca2+ channels (LTCCs) with fundamentally different gating properties; in addition to LTCCs resembling cardiac channels, LTCCs with anomalous gating were identified in a variety of neurons, including cerebellar granule cells. Anomalous LTCC gating is mainly characterized by long reopenings after repolarization following strong depolarizations or trains of action potentials. To elucidate the unknown molecular nature of anomalous LTCCs, we performed single-channel patch-clamp recordings from cerebellar granule cells of wild-type, Ca(v)1.3-/- and Ca(v)1.2DHP-/- [containing a mutation in the Ca(v)1.2 alpha1 subunit that eliminates dihydropyridine (DHP) sensitivity] mice. Quantitative reverse transcription-PCR revealed that Ca(v)1.2 accounts for 89% and Ca(v)1.3 for 11% of the LTCC transcripts in wild-type cerebellar granule cells, whereas Ca(v)1.1 and Ca(v)1.4 are expressed at insignificant levels. Anomalous LTCCs were observed in neurons of Ca(v)1.3-/- mice with a frequency not different from wild type. In the presence of the DHP agonist (+)-(S)-202-791, the typical prepulse-induced reopenings of anomalous LTCCs after repolarization were shorter in Ca(v)1.2DHP-/- neurons than in Ca(v)1.3-/- neurons. Reopenings in Ca(v)1.2DHP-/- neurons in the presence of the DHP agonist were similar to those in wild-type neurons in the absence of the agonist. These data show that Ca(v)1.2alpha1 subunits are the pore forming subunits of anomalous LTCCs in mouse cerebellar granule cells. Given the evidence that Ca(v)1.2 channels are specifically involved in sustained Ras-MAPK (mitogen-activated protein kinase)-dependent cAMP response element-binding protein phosphorylation and LTCC-dependent hippocampal long-term potentiation (LTP) (Moosmang et al., 2005), we discuss the hypothesis that anomalous rather than cardiac-type Ca(v)1.2 channels are specifically involved in LTCC-dependent and gene transcription-dependent LTP. PMID- 17409249 TI - GABAergic control of adult hippocampal neurogenesis in relation to behavior indicative of trait anxiety and depression states. AB - Stressful experiences in early life are known risk factors for anxiety and depressive illnesses, and they inhibit hippocampal neurogenesis and the expression of GABA(A) receptors in adulthood. Conversely, deficits in GABAergic neurotransmission and reduced neurogenesis are implicated in the etiology of pathological anxiety and diverse mood disorders. Mice that are heterozygous for the gamma2 subunit of GABA(A) receptors exhibit a modest functional deficit in mainly postsynaptic GABA(A) receptors that is associated with a behavioral, cognitive, and pharmacological phenotype indicative of heightened trait anxiety. Here we used cell type-specific and developmentally controlled inactivation of the gamma2 subunit gene to further analyze the mechanism and brain substrate underlying this phenotype. Heterozygous deletion of the gamma2 subunit induced selectively in immature neurons of the embryonic and adult forebrain resulted in reduced adult hippocampal neurogenesis associated with heightened behavioral inhibition to naturally aversive situations, including stressful situations known to be sensitive to antidepressant drug treatment. Reduced adult hippocampal neurogenesis was associated with normal cell proliferation, indicating a selective vulnerability of postmitotic immature neurons to modest functional deficits in gamma2 subunit-containing GABA(A) receptors. In contrast, a comparable forebrain-specific GABA(A) receptor deficit induced selectively in mature neurons during adolescence lacked neurogenic and behavioral consequences. These results suggest that modestly reduced GABA(A) receptor function in immature neurons of the developing and adult brain can serve as a common molecular substrate for deficits in adult neurogenesis and behavior indicative of anxious and depressive-like mood states. PMID- 17409251 TI - The cortical representation of objects rotating in depth. AB - The perception of motion provides valuable interpolations of the visual scene. This fundamental capacity of the visual system is evident in apparent rotation: by presenting only two images of an object rotated in space, a vivid illusion of a smooth apparent motion in three dimensions can be induced. The unseen interpolated rotation views are filled in by the visual system. In the present study, we identified the cortical network responsible for this filling-in process. We argue that cross talk between areas of the ventral and dorsal visual pathways promote the illusion of smooth apparent rotation. Most interestingly, the network represents the unseen object views. Using functional magnetic resonance adaptation, we are able to show that the cortical network selectively adapts to the illusory object views. Our findings provide strong evidence for cortical representations of three-dimensional rotating objects that are view invariant with respect to the rotation path. Furthermore, our results confirm psychophysical investigations that unseen interpolated rotation views can be primed by apparent motion. By applying functional magnetic resonance adaptation, we show for the first time cortical adaptation to unseen objects. Together, our neuroimaging study advances the understanding of the cortical mechanisms mediating the influence of motion on object processing. PMID- 17409252 TI - Both doublecortin and doublecortin-like kinase play a role in cortical interneuron migration. AB - Type I lissencephaly, a genetic disease characterized by disorganized cortical layers and gyral abnormalities, is associated with severe cognitive impairment and epilepsy. Two genes, LIS1 and doublecortin (DCX), have been shown to be responsible for a large proportion of cases of type I lissencephaly. Both genes encode microtubule-associated proteins that have been shown to be important for radial migration of cortical pyramidal neurons. To investigate whether DCX also plays a role in cortical interneuron migration, we inactivated DCX in the ganglionic eminence of rat embryonic day 17 brain slices using short hairpin RNA. We found that, when DCX expression was blocked, the migration of interneurons from the ganglionic eminence to the cerebral cortex was slowed but not absent, similar to what had previously been reported for radial neuronal migration. In addition, the processes of DCX-deficient migrating interneurons were more branched than their counterparts in control experiments. These effects were rescued by DCX overexpression, confirming the specificity to DCX inactivation. A similar delay in interneuron migration was observed when Doublecortin-like kinase (DCLK), a microtubule-associated protein related to DCX, was inactivated, although the morphology of the cells was not affected. The importance of these genes in interneuron migration was confirmed by our finding that the cortices of Dcx, Dclk, and Dcx/Dclk mutant mice contained a reduced number of such cells in the cortex and their distribution was different compared with wild-type controls. However, the defect was different for each group of mutant animals, suggesting that DCX and DCLK have distinct roles in cortical interneuron migration. PMID- 17409253 TI - Rac1 controls the formation of midline commissures and the competency of tangential migration in ventral telencephalic neurons. AB - Previous studies using dominant-mutant constructs have implicated Rac1 GTPase in neuritogenesis and neuronal migration. However, overexpression of dominant mutants generally blocks Rho-GTPase activity; thus, it may not reveal the specific or physiological functions of Rac1. To address this issue, we have applied a conditional gene-targeting strategy, using Foxg1-Cre mice to delete Rac1 in the ventricular zone (VZ) of telencephalon and Dlx5/6-Cre-IRES (internal ribosomal entry site)-EGFP (enhanced green fluorescent protein) (Dlx5/6-CIE) in the subventricular zone (SVZ) of ventral telencephalon, respectively. Surprisingly, the deletion of Rac1 in VZ progenitors did not prevent axonal outgrowth of telencephalic neurons. However, the anterior commissure was absent, and the corpus callosal as well as hippocampal commissural axons failed to cross the midline in Rac1/Foxg1-Cre knock-out embryos. The thalamocortical and corticothalamic axons also showed defasciculation or projection defects. These results suggest that Rac1 controls axon guidance rather than neuritogenesis. In addition, although Rac1/Foxg1-Cre knock-out embryos showed delayed radial migration of cortical projection neurons and severe impairment of tangential migration by the ventral telencephalon-derived interneurons, deletion of Rac1 in the SVZ by Dlx5/6-CIE mice produced no discernible defects in tangential migration. These contrasting effects of Rac1 deletion on tangential migration suggest that Rac1 is dispensable for cellular motility per se during neuronal migration. Together, these results underscore the challenge of deciphering the biological functions of Rac1, and Rho-GTPases in general, during mammalian brain development. Moreover, they indicate that Rac1 has a critical role in axon guidance and in acquisition of migratory competency during differentiation of the progenitors for the ventral telencephalon-derived interneurons. PMID- 17409254 TI - Choice of contact points during multidigit grasping: effect of predictability of object center of mass location. AB - It has been shown that when subjects can predict object properties [e.g., weight or center of mass (CM)], fingertip forces are appropriately scaled before the object is lifted, i.e., before somatosensory feedback can be processed. However, it is not known whether subjects, in addition to these anticipatory force mechanisms, exploit the ability to choose where digits can be placed to facilitate object manipulation. We addressed this question by asking subjects to reach and grasp an object whose CM was changed to the left, center, or right of the object in either a predictable or unpredictable manner. The only task requirement was to minimize object roll during lift. We hypothesized that subjects would modulate contact points but only when object CM location could be predicted. As expected, object roll was significantly smaller in the predictable condition. This experimental condition was also associated with statistically distinct spatial distributions of contact points as a function of object CM location but primarily when large torques had to be counteracted, i.e., for right and left CM locations. In contrast, when subjects could not anticipate CM location, a "default" distribution of contact points was used, this being statistically indistinguishable from that adopted for the center CM location in the predictable condition. We conclude that choice of contact points is integrated with anticipatory force control mechanisms to facilitate object manipulation. These results demonstrate that planning of digit placement is an important component of grasp control. PMID- 17409255 TI - A new code for contrast in the primate visual pathway. AB - We characterize a hitherto undocumented type of neuron present in the regions bordering the principal layers of the macaque lateral geniculate nucleus. Neurons of this type were distinguished by a high and unusually regular maintained discharge that was suppressed by spatiotemporal modulation of luminance or chromaticity within the receptive field. The response to any effective stimulus was a reduction in discharge, reminiscent of the "suppressed-by-contrast" cells of the cat retina. To a counterphase-modulated grating, the response was a phase insensitive suppression modulated at twice the stimulus frequency, implying a receptive field comprised of multiple mechanisms that generate rectifying responses. This distinctive nonlinearity makes the neurons well suited to computing a measure of contrast energy; such a signal might be important in regulating sensitivity early in visual cortex. PMID- 17409256 TI - Ipsilateral whiskers suppress experience-dependent plasticity in the barrel cortex. AB - Each cerebral hemisphere processes sensory input from both sides of the body, but the impact of this convergence on shaping and modifying receptive field properties remains controversial. Here we investigated the effect of chronic deprivation of ipsilateral sensory whiskers on receptive field plasticity in primary somatosensory cortex. In the absence of ipsilateral whiskers, cortical receptive fields were significantly larger than control after 1 week. Removal of all but a single whisker from one side of the face [single-whisker experience (SWE)] has been shown to result in the expansion of the cortical area responding to the spared whisker. We compared the effects of SWE in the presence (SWE unilateral) and absence (SWE-bilateral) of ipsilateral whiskers. SWE-bilateral deprivation results in a significant increase in neuronal responses to spared whisker stimulation both in its cognate barrel column and in adjacent, surrounding barrel columns compared with control and SWE-unilateral deprived animals. Surround receptive fields in deprived columns were maintained in SWE bilateral treated animals but depressed in SWE-unilateral animals. The increase in spared whisker responses was progressive with longer deprivation periods. These data show that ipsilateral whiskers can constrain receptive field size in the barrel cortex. PMID- 17409257 TI - Plexin-B2 controls the development of cerebellar granule cells. AB - Cerebellar granule cell progenitors proliferate postnatally in the upper part of the external granule cell layer (EGL) of the cerebellum. Postmitotic granule cells differentiate and migrate, tangentially in the EGL and then radially through the molecular and Purkinje cell layers. The molecular control of the transition between proliferation and differentiation in cerebellar granule cells is poorly understood. We show here that the transmembrane receptor Plexin-B2 is expressed by proliferating granule cell progenitors. To study Plexin-B2 function, we generated a targeted mutation of mouse Plexin-B2. Most Plexin-B2(-/-) mutants die at birth as a result of neural tube closure defects. Some mutants survive but their cerebellum cytoarchitecture is profoundly altered. This is correlated with a disorganization of the timing of granule cell proliferation and differentiation in the EGL. Many differentiated granule cells migrate inside the cerebellum and keep proliferating. These results reveal that Plexin-B2 controls the balance between proliferation and differentiation in granule cells. PMID- 17409258 TI - Activation of 5-HT1A receptors in the paragigantocellularis lateralis decreases shivering during cooling in the conscious piglet. AB - Activation of 5-HT(1A) receptors in the medullary raphe decreases sympathetic outflow to thermoregulatory mechanisms, including brown adipose tissue (BAT), thermogenesis, and peripheral vasoconstriction when these mechanisms are previously activated with leptin, prostaglandins, or cooling. These same mechanisms are also inhibited during rapid eye movement (REM) sleep. It is not known whether shivering is also modulated by medullary raphe neurons. We previously showed in the conscious piglet that activation of 5-HT(1A) receptors with 8-OH-DPAT (DPAT) in the paragigantocellularis lateralis (PGCL), a medullary region lateral to the midline raphe that contains 5-HT neurons, decreases heart rate, body temperature and muscle activity during non-rapid eye movement (NREM) sleep. We therefore hypothesized that activation of 5-HT(1A) receptors in the PGCL would also attenuate shivering and peripheral vasoconstriction during cooling. During REM sleep in a cool environment, shivering, carbon dioxide production, and body temperature decreased, and ear capillary blood flow and ear skin temperature increased. Shivering associated with rapid cooling was attenuated after dialysis of DPAT into the PGCL. In animals maintained in a continuously cool environment, dialysis of DPAT into the PGCL attenuated shivering and decreased body temperature, but there were no significant increases in ear capillary blood flow or ear skin temperature. We conclude that both naturally occurring REM sleep and exogenous activation of 5-HT(1A) receptors in the PGCL are associated with a suspension of shivering during cooling. Our data are consistent with the hypothesis that 5-HT neurons in the PGCL facilitate oscillating spinal motor circuits involved in shivering but are less involved in modulating sympathetically mediated thermoregulatory mechanisms. PMID- 17409259 TI - AMP does not induce torpor. AB - Torpor, a state characterized by a well-orchestrated reduction of metabolic rate and body temperature (T(b)), is employed for energetic savings by organisms throughout the animal kingdom. The nucleotide AMP has recently been purported to be a primary regulator of torpor in mice, as circulating AMP is elevated in the fasted state, and administration of AMP causes severe hypothermia. However, we have found that the characteristics and parameters of the hypothermia induced by AMP were dissimilar to those of fasting-induced torpor bouts in mice. Although administration of AMP induced hypothermia (minimum T(b) = 25.2 +/- 0.6 degrees C) similar to the depth of fasting-induced torpor (24.9 +/- 1.5 degrees C), ADP and ATP were equally effective in lowering T(b) (minimum T(b): 24.8 +/- 0.9 degrees C and 24.0 +/- 0.5 degrees C, respectively). The maximum rate of T(b) fall into hypothermia was significantly faster with injection of adenine nucleotides (AMP: 0.24 +/- 0.03; ADP: -0.24 +/- 0.02; ATP: -0.25 +/- 0.03 degrees C/min) than during fasting-induced torpor (-0.13 +/- 0.02 degrees C/min). Heart rate decreased from 755 +/- 15 to 268 +/- 17 beats per minute (bpm) within 1 min of AMP administration, unlike that observed during torpor (from 646 +/- 21 to 294 +/ 19 bpm over 35 min). Finally, the hypothermic effect of AMP was blunted with preadministration of an adenosine receptor blocker, suggesting that AMP action on T(b) is mediated via the adenosine receptor. These data suggest that injection of adenine nucleotides into mice induces a reversible hypothermic state that is unrelated to fasting-induced torpor. PMID- 17409260 TI - Renal responses to the kappa-opioid-receptor agonist U-50488H in conscious lambs. AB - In adult animals and humans, activation of kappa-opioid receptors results in a diuresis. The aim of the present study was to investigate whether kappa-opioids are also diuretic early in life and whether this is altered during postnatal maturation. Therefore, the renal effects of the kappa-opioid-receptor agonist U 50488H were measured in two separate age groups of conscious lambs at two stages of postnatal maturation (approximately 1 wk and approximately 6 wk) under physiological conditions. To evaluate whether the renal responses to U-50488H resulted from receptor-dependent effects, responses to U-50488H were also tested in the presence of the specific kappa-opioid-receptor antagonist 5' guanidinonaltrindole (GNTI). Urinary flow rate, free water clearance, and electrolyte excretions and clearances were measured for 30 min before and for 90 min after intravenous injection of U-50488H or vehicle. An increase in urinary flow rate accompanied by an increase in free water clearance occurred in response to administration of U-50488H but not vehicle. There were no effects of U-50488H on electrolyte excretions or clearances at either 1 or 6 wk of postnatal life. Although there were no effects of GNTI on any of the measured or calculated variables, the aforementioned diuretic response to U-50488H was abolished by pretreatment with GNTI in both age groups. We conclude that kappa-opioid receptors are diuretic early in life and that this response does not appear to be altered as postnatal maturation proceeds. Therefore, these data provide evidence that activation of kappa-opioid receptors early in life may lead to alterations in fluid balance. PMID- 17409261 TI - Phosphorylation of extracellular signal-regulated kinases in urinary bladder in rats with cyclophosphamide-induced cystitis. AB - Phosphorylated ERK expression has been demonstrated in the central and peripheral nervous system after various stimuli, including visceral stimulation. Changes in the activation (i.e., phosphorylation) of extracellular signal-regulated kinases (pERK) were examined in the urinary bladder after 4 h (acute), 48 h (intermediate), or chronic (10 day) cyclophosphamide (CYP) treatment. CYP-induced cystitis significantly (P < or = 0.01) increased pERK expression in the urinary bladder with intermediate (48 h) and chronic CYP treatment. Immunohistochemistry for pERK immunoreactivity revealed little pERK-IR in control or acute (4 h) CYP treated rat urinary bladders. However, pERK expression was significantly (P < or = 0.01) upregulated in the urothelium after 48 h or chronic CYP treatment. Whole mount preparations of urothelium/lamina propria or detrusor smooth muscle from control (noninflamed) rats showed no pERK-IR in PGP9.5-labeled nerve fibers in the suburothelial plexus. However, with CYP-treatment (48 h, chronic), a few pERK IR nerve fibers in the suburothelial plexus of whole mount preparations of bladder and at the serosal edge of urinary bladder sections were observed. pERK IR cells expressing the CD86 antigen were also observed in urinary bladder from CYP-treated rats (48 h, chronic). Treatment with the upstream inhibitor of ERK phosphorylation, U0126, significantly (P < or= 0.01) increased bladder capacity in CYP-treated rats (48 h). These studies suggest that therapies targeted at pERK pathways may improve urinary bladder function in CYP-treated rats. PMID- 17409262 TI - Role of midbrain in the control of breathing in anuran amphibians. AB - The present study was designed to explore systematically the midbrain of unanesthetized, decerebrate anuran amphibians (bullfrogs), using chemical and electrical stimulation and midbrain transections to identify sites capable of exciting and inhibiting breathing. Ventilation was measured as fictive motor output from the mandibular branch of the trigeminal nerve and the laryngeal branch of the vagus nerve. The results of our transection studies suggest that, under resting conditions, the net effect of inputs from sites within the rostral half of the midbrain is to increase fictive breathing frequency, whereas inputs from sites within the caudal half of the midbrain have no net effect on fictive breathing frequency but appear to act on the medullary central rhythm generator to produce episodic breathing. The results of our stimulation experiments indicate that the principal sites in the midbrain that are capable of exciting or inhibiting the fictive frequency of lung ventilation, and potentially clustering breaths into episodes, appear to be those primarily involved in visual and auditory integration, motor functions, and attentional state. PMID- 17409263 TI - Glucose supplements increase human muscle in vitro Na+-K+-ATPase activity during prolonged exercise. AB - Regulation of maximal Na(+)-K(+)-ATPase activity in vastus lateralis muscle was investigated in response to prolonged exercise with (G) and without (NG) oral glucose supplements. Fifteen untrained volunteers (14 males and 1 female) with a peak aerobic power (Vo(2)(peak)) of 44.8 +/- 1.9 ml.kg(-1).min(-1); mean +/- SE cycled at approximately 57% Vo(2)(peak) to fatigue during both NG (artificial sweeteners) and G (6.13 +/- 0.09% glucose) in randomized order. Consumption of beverage began at 30 min and continued every 15 min until fatigue. Time to fatigue was increased (P < 0.05) in G compared with NG (137 +/- 7 vs. 115 +/- 6 min). Maximal Na(+)-K(+)-ATPase activity (V(max)) as measured by the 3-O methylfluorescein phosphatase assay (nmol.mg(-1).h(-1)) was not different between conditions prior to exercise (85.2 +/- 3.3 or 86.0 +/- 3.9), at 30 min (91.4 +/- 4.7 vs. 91.9 +/- 4.1) and at fatigue (92.8 +/- 4.3 vs. 100 +/- 5.0) but was higher (P < 0.05) in G at 90 min (86.7 +/- 4.2 vs. 109 +/- 4.1). Na(+)-K(+) ATPase content (beta(max)) measured by the vanadate facilitated [(3)H]ouabain binding technique (pmol/g wet wt) although elevated (P < 0.05) by exercise (0<30, 90, and fatigue) was not different between NG and G. At 60 and 90 min of exercise, blood glucose was higher (P < 0.05) in G compared with NG. The G condition also resulted in higher (P < 0.05) serum insulin at similar time points to glucose and lower (P < 0.05) plasma epinephrine and norepinephrine at 90 min of exercise and at fatigue. These results suggest that G results in an increase in V(max) by mechanisms that are unclear. PMID- 17409264 TI - Spontaneous sleep and homeostatic sleep regulation in ghrelin knockout mice. AB - Ghrelin is well known for its feeding and growth hormone-releasing actions. It may also be involved in sleep regulation; intracerebroventricular administration and hypothalamic microinjections of ghrelin stimulate wakefulness in rats. Hypothalamic ghrelin, together with neuropeptide Y and orexin form a food intake regulatory circuit. We hypothesized that this circuit also promotes arousal. To further investigate the role of ghrelin in the regulation of sleep-wakefulness, we characterized spontaneous and homeostatic sleep regulation in ghrelin knockout (KO) and wild-type (WT) mice. Both groups of mice exhibited similar diurnal rhythms with more sleep and less wakefulness during the light period. In ghrelin KO mice, spontaneous wakefulness and rapid-eye-movement sleep (REMS) were slightly elevated, and non-rapid-eye-movement sleep (NREMS) was reduced. KO mice had more fragmented NREMS than WT mice, as indicated by the shorter and greater number of NREMS episodes. Six hours of sleep deprivation induced rebound increases in NREMS and REMS and biphasic changes in electroencephalographic slow wave activity (EEG SWA) in both genotypes. Ghrelin KO mice recovered from NREMS and REMS loss faster, and the delayed reduction in EEG SWA, occurring after sleep loss-enhanced increases in EEG SWA, was shorter-lasting compared with WT mice. These findings suggest that the basic sleep-wake regulatory mechanisms in ghrelin KO mice are not impaired and they are able to mount adequate rebound sleep in response to a homeostatic challenge. It is possible that redundancy in the arousal systems of the brain or activation of compensatory mechanisms during development allow for normal sleep-wake regulation in ghrelin KO mice. PMID- 17409265 TI - Effects of acute and chronic sleep loss on immune modulation of rats. AB - Sleep deprivation is now recognized as an increasingly common condition inherent to modern society, and one that in many ways, is detrimental to certain physiological systems, namely, immune function. Although sleep is now viewed by a significant body of researchers as being essential for the proper working of a host of defense systems, the consequences of a lack of sleep on immune function remains to be fully comprehended. The aim of the current study was to investigate how paradoxical sleep deprivation (PSD) for 24 and 96 h and sleep restriction (SR) for 21 days by the modified multiple-platform method, and their respective 24-h recovery periods, affect immune activation in rats. To this end, we assessed circulating white blood cell counts, lymphocyte count within immune organs, as well as Ig and complement production. The data revealed that PSD for 96 h increased complement C3 and corticosterone concentration in relation to the control group. In contrast, the spleen weight, total leukocytes, and lymphocytes decreased during SR for 21 days when compared with the control group, although production of a certain class of immunoglobulin, the IgM, did increase. After recovery sleep, lymphocyte count in axillary lymph nodes grew when rats had rebound sleep after PSD for 24 h, neutrophils increased after PSD 96 h and lymphocytes numbers were higher after SR 21 days. Such alterations during sleep deprivation suggest only minor alterations of nonspecific immune parameters during acute PSD, and a significant impairment in cellular response during chronic SR. PMID- 17409266 TI - Differential body weight and feeding responses to high-fat diets in rats and mice lacking cholecystokinin 1 receptors. AB - Prior data demonstrated differential roles for cholecystokinin (CCK)1 receptors in maintaining energy balance in rats and mice. CCK1 receptor deficiency results in hyperphagia and obesity of Otsuka Long-Evans Tokushima Fatty (OLETF) rats but not in mice. To ascertain the role of CCK1 receptors in high-fat-diet (HFD) induced obesity, we compared alterations in food intake, body weight, fat mass, plasma glucose, and leptin levels, and patterns of hypothalamic gene expression in OLETF rats and mice lacking CCK1 receptors in response to a 10-wk exposure to HFD. Compared with Long-Evans Tokushima Otsuka (LETO) control rats, OLETF rats on HFD had sustained overconsumption over the 10-wk period. High fat feeding resulted in greater increases in body weight and plasma leptin levels in OLETF than in LETO rats. In situ hybridization determinations revealed that, while HFD reduced neuropeptide Y (NPY) mRNA expression in both the arcuate nucleus (Arc) and the dorsomedial hypothalamus (DMH) of LETO rats, HFD resulted in decreased NPY expression in the Arc but not in the DMH of OLETF rats. In contrast to these results in OLETF rats, HFD increased food intake and induced obesity to an equal degree in both wild-type and CCK1 receptor(-/-) mice. NPY gene expression was decreased in the Arc in response to HFD, but was not detectable in the DMH in both wild-type and CCK1 receptor(-/-) mice. Together, these data provide further evidence for differential roles of CCK1 receptors in the controls of food intake and body weight in rats and mice. PMID- 17409267 TI - Alternative splicing due to an intronic SNP in HMSD generates a novel minor histocompatibility antigen. AB - Here we report the identification of a novel human leukocyte antigen (HLA)-B44 restricted minor histocompatibility antigen (mHA) with expression limited to hematopoietic cells. cDNA expression cloning studies demonstrated that the cytotoxic T lymphocyte (CTL) epitope of interest was encoded by a novel allelic splice variant of HMSD, hereafter designated as HMSD-v. The immunogenicity of the epitope was generated by differential protein expression due to alternative splicing, which was completely controlled by 1 intronic single-nucleotide polymorphism located in the consensus 5' splice site adjacent to an exon. Both HMSD-v and HMSD transcripts were selectively expressed at higher levels in mature dendritic cells and primary leukemia cells, especially those of myeloid lineage. Engraftment of mHA(+) myeloid leukemia stem cells in nonobese diabetic/severe combined immunodeficient (NOD/SCID)/gammac(null) mice was completely inhibited by in vitro preincubation with the mHA-specific CTL clone, suggesting that this mHA is expressed on leukemic stem cells. The patient from whom the CTL clone was isolated demonstrated a significant increase of the mHA-specific T cells in posttransplantation peripheral blood, whereas mHA-specific T cells were undetectable in pretransplantation peripheral blood and in peripheral blood from his donor. These findings suggest that the HMSD-v-encoded mHA (designated ACC-6) could serve as a target antigen for immunotherapy against hematologic malignancies. PMID- 17409268 TI - Basophils are essential initiators of a novel type of chronic allergic inflammation. AB - Basophils represent less than 1% of peripheral blood leukocytes and have often been considered as minor and possibly redundant circulating mast cells. The recent finding that basophils readily generate large quantities of T helper 2 (Th2) cytokines such as IL-4 provided new insights into the possible role of basophils in allergic disorders and immunity to pathogens. However, in-depth studies on basophils, particularly their functions in vivo, have been hampered by the lack of appropriate animal models, such as mutant animals deficient only in basophils. Here, we established a mAb that reacted with mouse basophils and depleted them when administered in vivo. The mAb treatment of mice did not show any significant effect on classical allergic reactions such as passive cutaneous anaphylaxis and contact hypersensitivity. In contrast, it completely abolished the development of IgE-mediated chronic allergic dermatitis that is characterized by massive eosinophil infiltration, even though basophils accounted for only approximately 2% of the infiltrates. The treatment during the progression of the dermatitis resulted in drastic reduction in numbers of infiltrating eosinophils and neutrophils, concomitantly with elimination of basophils from the skin lesions. Thus, basophils play a pivotal role in the development of IgE-mediated chronic allergic inflammation, as an initiator rather than as an effector. PMID- 17409269 TI - Sickle cell trait and the risk of venous thromboembolism among blacks. AB - People with sickle cell disease have a chronically activated coagulation system and display hemostatic perturbations, but it is unknown whether they experience an increased risk of venous thromboembolism. We conducted a case-control study of venous thromboembolism that included 515 hospitalized black patients and 555 black controls obtained from medical clinics. All subjects were assayed for hemoglobin S and hemoglobin C genotypes. The prevalence of the S allele was 0.070 and 0.032 for case patients and controls, respectively (P < .001). The odds that a patient had sickle cell trait were approximately twice that of a control, indicating that the risk of venous thromboembolism is increased approximately 2 fold among blacks with sickle cell trait compared with those with the wild-type genotype (odds ratio = 1.8 with 95% confidence interval, 1.2-2.9). The odds ratio for pulmonary embolism and sickle cell trait was higher, 3.9 (2.2-6.9). The prevalence of sickle cell disease was also increased among case patients compared with controls. We conclude that sickle cell trait is a risk factor for venous thromboembolism and that the proportion of venous thromboembolism among blacks attributable to the mutation is approximately 7%. PMID- 17409270 TI - Natural killer cell-derived human granzyme H induces an alternative, caspase independent cell-death program. AB - Granzyme H (GzmH) belongs to a family of 5 human serine proteases that are expressed by cytotoxic immune effector cells. Although GzmH is most closely related to the caspase-activating granzyme B (GzmB), neither a natural substrate nor a role in immune defense reactions has been demonstrated for this orphan granzyme. In rodents, multiple related genes exist, but none of these can be regarded as functional homologs. Here we show that host cells are efficiently killed by GzmH after perforin and streptolysin O-mediated delivery into the cytosol. Dying cells show typical hallmarks of programmed cell death, such as mitochondrial depolarization, reactive oxygen species (ROS) generation, DNA degradation, and chromatin condensation. Contrary to GzmB, cell death by GzmH does not involve the activation of executioner caspases, the cleavage of Bid or inhibitor of caspase-activated DNase (ICAD), or the release of cytochrome c. The high expression levels of GzmH in naive natural killer (NK) cells and its potent killing ability strongly support the role of the protease in triggering an alternative cell-death pathway in innate immunity. PMID- 17409271 TI - Possible bioactivation pathways of lamotrigine. AB - The anticonvulsant lamotrigine is associated with idiosyncratic drug reactions, especially skin rashes. Most idiosyncratic reactions are believed to be caused by reactive metabolites. Previous studies have found evidence that an arene oxide is formed in rats; however, when we incubated radiolabeled lamotrigine with rat liver microsomes virtually no covalent binding was detected, and the expected downstream phenolic metabolites are not observed in humans. Rare cases of agranulocytosis have been associated with lamotrigine therapy, and we found that lamotrigine is oxidized to two different N-chloro products by HOCl. The more reactive N-chloro metabolite forms an adduct with N-acetylhistidine, and covalent binding was observed when radiolabeled lamotrigine was incubated with myeloperoxidase/H(2)O(2)/Cl(-). Another lamotrigine metabolite is an N-oxide. If this N-oxide were sulfated, it might be sufficiently reactive to bind to protein. The synthetic N-sulfate reacted with N-acetylserine; however, no covalent binding was detected when the radiolabeled N-oxide was incubated with sulfotransferase. We also investigated the possibility that lamotrigine might be oxidized to a free radical by other peroxidases or oxidized by other enzymes such as prostaglandin H synthase or tyrosinase, but no evidence of oxidation was found, and lamotrigine did not cause any detectable increase in lipid peroxidation in vivo. In view of the virtual lack of covalent binding to hepatic microsomes and the lack of any other likely pathway leading to metabolic activation in the skin, it is possible that the parent drug rather than a reactive metabolite causes lamotrigine-induced skin rashes. PMID- 17409272 TI - Nateglinide and mitiglinide, but not sulfonylureas, induce insulin secretion through a mechanism mediated by calcium release from endoplasmic reticulum. AB - Nateglinide and mitiglinide (glinides) are characterized as rapid-onset and short acting insulinotropic agents. Although both compounds do not have a sulfonylurea structure, it has been postulated that insulin secretion is preceded by their binding to Kir6.2/SUR1 complex, and a mechanism of insulin secretion of glinides has been accounted for by this pathway. However, we hypothesized the involvement of additional mechanisms of insulin secretion enhanced by glinides, and we analyzed the pattern of time course of insulin secretion from MIN6 cells with the existence of agents that have specific pharmacologic actions. Dose-dependent effects of tolbutamide, glibenclamide, nateglinide, and mitiglinide were observed. Insulin secretion induced by 3 microM tolbutamide and 1 nM glibenclamide was completely inhibited by 10 microM diazoxide and 3 microM verapamil, although the latter half-component of insulin secretion profile induced by 3 microM nateglinide or 30 nM mitiglinide remained with the existence of those agents. Glinides enhanced insulin secretion even in Ca2+-depleted medium, and its pattern of secretion was same as the pattern with existence of verapamil. The latter half was suppressed by 1 microM dantrolene, and concomitant addition of verapamil and dantrolene completely suppressed the entire pattern of insulin secretion enhanced by nateglinide. Thus, we conclude that glinide action is demonstrated through two pathways, dependently and independently, from the pathway through K(ATP) channels. We also demonstrated that the latter pathway involves the intracellular calcium release from endoplasmic reticulum via ryanodine receptor activation. PMID- 17409273 TI - Chronic treatment with carvedilol improves Ca(2+)-dependent ATP consumption in triton X-skinned fiber preparations of human myocardium. AB - Evidence is given that beta-blocker treatment differentially influences gene expression and up-regulation of beta(1)-adrenoceptors in human myocardium. Here, we investigate whether long-term treatment with carvedilol or metoprolol may functionally alter myofibrillar function in end-stage human heart failure. Investigations were performed in Triton X (1%, 4 degrees C, 20 h)-skinned fiber preparations of explanted hearts from patients undergoing heart transplantation due to idiopathic dilative cardiomyopathy. Five patients were not on beta adrenoceptor blocker treatment (DCM_NBB), and 5 patients received either carvedilol (DCM_CAR) or metoprolol (DCM_MET). Nonfailing (NF) donor hearts (n = 5), which could not be transplanted due to technical reasons, were investigated for comparison. Ca(2+)-dependent tension (DT) development and actomyosin-ATPase activity (MYO) were measured and tension-dependent ATP consumption was calculated by the ratio of DT and MYO ("tension cost"). In addition, we measured the phosphorylation of troponin I (TNI) by back phosphorylation. Maximal DT and TNI phosphorylation were reduced, with myofibrillar Ca(2+) sensitivity of DT and MYO as well as tension cost being increased in DCM_NBB compared with NF. Metoprolol treatment restored TNI phosphorylation, decreased Ca(2+) sensitivity of tension development and of myosin-ATPase activity, but did not alter the tension dependent ATP consumption. Carvedilol treatment improved maximal DT and significantly decreased tension-dependent ATP consumption without altering myofibrillar Ca(2+) sensitivity. TNI dephosphorylation was increased in patients treated with carvedilol. In conclusion, chronic beta-adrenoceptor blockade functionally alters myofibrillar function. The more economic cross-bridge cycling in patients under carvedilol treatment may provide an explanation for the efficacy of carvedilol in the treatment of chronic heart failure patients. PMID- 17409274 TI - Development of a Transcreener kinase assay for protein kinase A and demonstration of concordance of data with a filter-binding assay format. AB - A Transcreener kinase fluorescence polarization (FP) assay has been developed for the serine/threonine kinase protein kinase A (PKA). The PKA Transcreener kinase assay is an homogenous, competitive antibody-based FP assay that uses Far Red Alexa Fluor 633-labeled adenosine 5' disphosphate (ADP) tracer and mouse monoclonal anti-ADP antibody. The Transcreener PKA assay was validated with both known PKA inhibitors and library compounds. The Transcreener PKA assay is resistant to low-wavelength (or common) fluorescent interference from small molecule library compounds and generates IC50 results comparable with current radioactive filter-binding assay. PMID- 17409275 TI - Cellular prion protein promotes proliferation and G1/S transition of human gastric cancer cells SGC7901 and AGS. AB - The function of cellular prion protein (PrP(C)), the essential protein for the pathogenesis and transmission of prion diseases, is still largely unknown. The putative roles of PrP(C) are thought to be related to cell signaling, survival, and differentiation. In a previous study, we showed that PrP(C) was overexpressed in gastric cancer tissues. In the present report, we show that ectopic expression of PrP(C) could promote tumorigenesis, proliferation, and G1/S transition in gastric cancer cells. Furthermore, CyclinD1, a protein related to cell cycle, was shown to be significantly up-regulated by PrP(C) at both mRNA and protein levels. PI3K/Akt pathway mediated above PrP(C) signal since PrP(C) increased the expression of phosphorylated Akt, and the specific inhibitor of Akt, LY294002, could markedly suppress growth of SGC7901 and transactivation of CyclinD1 induced by PrP(C). Octapeptide repeat region played a vital role in this function, as deletion of this region abolished or reduced these effects. Collectively, this study demonstrates that overexpression of PrP(C) might promote the tumorigenesis and proliferation of gastric cancer cells at least partially through activation of PI3K/Akt pathway and subsequent transcriptional activation of CyclinD1 to regulate the G1/S phase transition, in which octapeptide repeat region might be an indispensable region. PMID- 17409277 TI - Downregulation of renal TonEBP in hypokalemic rats. AB - Hypokalemia causes a significant decrease in the tonicity of the renal medullary interstitium in association with reduced expression of sodium transporters in the distal tubule. We asked whether hypokalemia caused downregulation of the tonicity responsive enhancer binding protein (TonEBP) transcriptional activator in the renal medulla due to the reduced tonicity. We found that the abundance of TonEBP decreased significantly in the outer and inner medullas of hypokalemic rats. Underlying mechanisms appeared different in the two regions because the abundance of TonEBP mRNA was lower in the outer medulla but unchanged in the inner medulla. Immunohistochemical examination of TonEBP revealed cell type-specific differences. TonEBP expression decreased dramatically in the outer and inner medullary collecting ducts, thick ascending limbs, and interstitial cells. In the descending and ascending thin limbs, TonEBP abundance decreased modestly. In the outer medulla, TonEBP shifted to the cytoplasm in the descending thin limbs. As expected, transcription of aldose reductase, a target of TonEBP, was decreased since the abundance of mRNA and protein was reduced. Downregulation of TonEBP appeared to have also contributed to reduced expression of aquaporin-2 and UT-A urea transporters in the renal medulla. In cultured cells, expression and activity of TonEBP were not affected by reduced potassium concentrations in the medium. These data support the view that medullary tonicity regulates expression and nuclear distribution of TonEBP in the renal medulla in cell type-specific manners. PMID- 17409276 TI - Crucial role of Rho-nuclear factor-kappaB axis in angiotensin II-induced renal injury. AB - This study was performed to determine the effectiveness of the Rho kinase inhibitor and NF-kappaB inhibitor in renal injury of ANG II-infused hypertensive rats. Male Sprague-Dawley rats, maintained on a normal diet, received either a sham operation (n = 7) or continuous ANG II infusion (120 ng/min) subcutaneously via minipumps. The ANG II-infused rats were further subdivided into three subgroups (n = 7 each) to receive one of the following treatments during the entire period: vehicle, Rho kinase inhibitor (fasudil; 3 mg.kg(-1).day(-1) ip), or NF-kappaB inhibitor (parthenolide; 1 mg.kg(-1).day(-1) ip). After 12 days of ANG II infusion, systolic blood pressure (BP; 208 +/- 7 vs. 136 +/- 3 mmHg), Rho kinase activity, NF-kappaB activity, renal ANG II contents (160 +/- 25 vs. 84 +/- 14 pg/g), monocytic chemotactic protein (MCP) 1 mRNA, interstitial macrophage infiltration, transforming growth factor-beta1 (TGF-beta1) mRNA, interstitial collagen-positive area, urinary protein excretion (43 +/- 6 vs. 11 +/- 2 mg/day), and urinary albumin excretion were significantly enhanced compared with the Sham group. While fasudil or parthenolide did not alter systolic BP (222 +/- and 190 +/- 21, respectively), both treatments completely blocked ANG II-induced enhancement of NF-kappaB activity, renal ANG II contents (103 +/- 11 and 116 +/- 21 pg/g, respectively), MCP1 mRNA, interstitial macrophage infiltration, TGF beta1 mRNA, interstitial collagen-positive area, urinary protein excretion (28 +/ 6 and 23 +/- 3 mg/day, respectively), and urinary albumin excretion. Importantly, parthenolide did not alter ANG II-induced Rho kinase activation although fasudil abolished ANG II-induced Rho kinase activation. These data indicate that the Rho-NF-kappaB axis plays crucial roles in the development of ANG II-induced renal injury independently from BP regulation. PMID- 17409278 TI - Activated protein C ameliorates LPS-induced acute kidney injury and downregulates renal INOS and angiotensin 2. AB - Endothelial dysfunction contributes significantly to acute renal failure (ARF) during inflammatory diseases including septic shock. Previous studies have shown that activated protein C (APC) exhibits anti-inflammatory properties and modulates endothelial function. Therefore, we investigated the effect of APC on ARF in a rat model of endotoxemia. Rats subjected to lipopolysaccharide (LPS) treatment exhibited ARF as illustrated by markedly reduced peritubular capillary flow and increased serum blood urea nitrogen (BUN) levels. Using quantitative two photon intravital microscopy, we observed that at 3 h post-LPS treatment, rat APC (0.1 mg/kg iv bolus) significantly improved peritubular capillary flow [288 +/- 15 microm/s (LPS) vs. 734 +/- 59 microm/s (LPS+APC), P = 0.0009, n = 6], and reduced leukocyte adhesion (P = 0.003) and rolling (P = 0.01) compared with the LPS-treated group. Additional experiments demonstrated that APC treatment significantly improved renal blood flow and reduced serum BUN levels compared with 24-h post-LPS treatment. Biochemical analysis revealed that APC downregulated inducible nitric oxide synthase (iNOS) mRNA levels and NO by products in the kidney. In addition, APC modulated the renin-angiotensin system by reducing mRNA expression levels of angiotensin-converting enzyme-1 (ACE1), angiotensinogen, and increasing ACE2 mRNA levels in the kidney. Furthermore, APC significantly reduced ANG II levels in the kidney compared with the LPS-treated group. Taken together, these data suggest that APC can suppress LPS-induced ARF by modulating factors involved in vascular inflammation, including downregulation of renal iNOS and ANG II systems. Furthermore, the data suggest a potential therapeutic role for APC in the treatment of ARF. PMID- 17409280 TI - Kinase signaling pathways regulate fine tuning of transporter activity. PMID- 17409279 TI - Thyroid hormone deficiency alters expression of acid-base transporters in rat kidney. AB - Hypothyroidism in humans is associated with incomplete distal renal tubular acidosis, presenting as the inability to respond appropriately to an acid challenge by excreting less acid. Here, we induced hypothyroidism in rats with methimazole (HYPO) and in one group substituted with l-thyroxine (EU). After 4 wk, acid-base status was similar in both groups. However, after 24 h acid loading with NH(4)Cl HYPO rats displayed a more pronounced metabolic acidosis. The expression of the Na(+)/H(+) exchanger NHE3, the Na(+)-phosphate cotransporter NaPi-IIa, and the B2 subunit of the vacuolar H(+)-ATPase was reduced in the brush border membrane of the proximal tubule of the HYPO group, paralleled by a lower abundance of the Na(+)/HCO(3)(-) cotransporter NBCe1 and a higher expression of the acid-secretory type A intercalated cell-specific Cl(-)/HCO(3)(-) exchanger AE1. In contrast to control conditions, the expression of NBCe1 was increased in the HYPO group during metabolic acidosis. In addition, net acid excretion was similar in both groups. The relative number of type A intercalated cells was increased in the connecting tubule and cortical collecting duct of the HYPO group during acidosis. Thus thyroid hormones modulate the renal response to an acid challenge and alter the expression of several key acid-base transporters. Mild hypothyroidism is associated only with a very mild defect in renal acid handling, which appears to be mainly located in the proximal tubule and is compensated by the distal nephron. PMID- 17409281 TI - Compromised renal microvascular reactivity of angiotensin type 1 double null mice. AB - Angiotensin type 1A (AT(1A)) and 1B (AT(1B)) receptor deletion (AT1DKO) results in renal microvascular disease, tubulointerstitial injury, and reduced blood pressure. To test the hypothesis that renal preglomerular responses to angiotensin (ANG) II are mediated by AT(1A) and AT(1B) receptors, experiments were performed in AT1DKO mice using the in vitro blood perfused juxtamedullary nephron technique. Kidneys were harvested from AT1DKO and wild-type (WT) mice and bathed with ANG II (1-100 nM), norepinephrine (NE; 100-1,000 nM), or acetylcholine (ACh; 10 microM). Baseline diameters of afferent (19.5 +/- 0.7 and 13.9 +/- 0.7 microm, n = 17 and 16) and efferent (15.5 +/- 2.1 and 10.8 +/- 1.0 microm, n = 4 and 7) arterioles of AT1DKO were significantly larger than WT. Afferent and efferent arteriolar responses to ANG II, 100, and 300 nM NE were absent in AT1DKO; although significant constriction to 1 microM NE was observed ( 17 +/- 5 and -23 +/- 6%, respectively). Afferent arterioles of WT mice dilated significantly in response to ACh (15.1 +/- 0.6 to 17.0 +/- 1.2 microm, n = 6); however, arterioles from AT1DKO tended to contract (19.9 +/- 1.2 to 17.8 +/- 1.6 microm; n = 6, P = 0.06). In summary, loss of ANG II-induced contraction, reduced vasoconstriction to NE, and endothelial cell dysfunction contribute to the renal vascular phenotype of AT1DKO mice. We conclude that ANG II signaling via the AT(1) receptor plays a pivotal role in basal renal microvascular tone and effectiveness to respond to vasoconstrictor and vasodilator agonists. PMID- 17409282 TI - NHE3 phosphorylation at serines 552 and 605 does not directly affect NHE3 activity. AB - Direct phosphorylation of sodium hydrogen exchanger type 3 (NHE3) is a well established physiological phenomenon; however, the exact role of NHE3 phosphorylation in its regulation remains unclear. The objective of this study was to evaluate whether NHE3 phosphorylation at serines 552 and 605 is physiologically regulated in vivo and, if so, whether changes in phosphorylation at these sites are tightly coupled to changes in transport activity. To this end, we directly compared PKA-induced NHE3 inhibition with site-specific changes in NHE3 phosphorylation in vivo and in vitro. In vivo, PKA was activated using an intravenous infusion of parathyroid hormone in Sprague-Dawley rats. In vitro, PKA was activated directly in opossum kidney (OKP) cells using forskolin and IBMX. NHE3 activity was assayed in microvillar membrane vesicles in the rat model and by (22)Na uptake in the OKP cell model. In both cases, NHE3 phosphorylation at serines 552 and 605 was determined using previously characterized monoclonal phosphospecific antibodies directed to these sites. In vivo, we found dramatic changes in NHE3 phosphorylation at serines 552 and 605 with PKA activation but no corresponding alteration in NHE3 activity. This dissociation between NHE3 phosphorylation and activity was further verified in OKP cells in which phosphorylation clearly preceded transport inhibition. We conclude that although phosphorylation of NHE3 at serines 552 and 605 is regulated by PKA both in vivo and in vitro, phosphorylation of these sites does not directly alter Na(+)/H(+) exchange activity. PMID- 17409283 TI - Localization of broadly selective equilibrative and concentrative nucleoside transporters, hENT1 and hCNT3, in human kidney. AB - Nucleoside transporters in kidney mediate renal reabsorption and secretion of nucleosides. Using RT-PCR, we demonstrated mRNAs encoding hENT1, hENT2, hCNT1, hCNT2, and hCNT3 in both cortex and medulla. Immunoblotting with crude membrane preparations revealed abundant hENT1 and hCNT3 in both cortex and medulla, and little, if any, hENT2, hCNT1, or hCNT2, indicating that the latter were either absent or below limits of detection of immunoassays. hENT1 immunostaining was observed on apical surfaces of proximal tubules and on both apical and basal surfaces of thick ascending loops of Henle and collecting ducts. Prominent hCNT3 immunostaining was observed on apical surfaces of proximal tubules and thick ascending loops of Henle in addition to some cytoplasmic staining. Equilibrium binding of [(3)H]nitrobenzylmercaptopurine ribonucleoside (NBMPR), a high affinity inhibitor of hENT1, to brush-border membrane vesicles from cortex confirmed the presence of hENT1 on apical surfaces of proximal tubules. Uptake of [(3)H]uridine by polarized renal proximal tubule cells exhibited a sodium dependent component that was inhibited by thymidine and inosine as well as a sodium-independent component that was partially inhibited by NBMPR and completely inhibited by dilazep, indicating high levels of hENT1 and hCNT3 and low levels of hENT2 activities. The presence of 1) transcripts for hENT1/2 and hCNT1/2/3 and the hENT1 and hCNT3 proteins in human kidneys and 2) hENT1, hENT2, and hCNT3 activities in cultured proximal tubule cells suggest involvement of hENT1, hCNT3, and possibly also hENT2 in renal handling of nucleosides and nucleoside drugs. PMID- 17409284 TI - Nigrostriatal damage preferentially decreases a subpopulation of alpha6beta2* nAChRs in mouse, monkey, and Parkinson's disease striatum. AB - Parkinson's disease is a neurodegenerative movement disorder characterized by a loss of substantia nigra dopamine neurons, and corresponding declines in molecular components present on striatal dopaminergic nerve terminals. These include the alpha6beta2(*) nicotinic acetylcholine receptors (nAChRs), which are localized exclusively on dopamine terminals in striatum ((*)denotes the presence of possible additional subunits). In this study, we used a novel alpha-conotoxin MII (alpha-CtxMII) analog E11A to further investigate alpha6beta2(*) nAChR subtypes in mouse, monkey, and human striatum. Receptor competition studies with (125)I-alpha-CtxMII showed that E11A inhibition curves were biphasic, suggesting the presence of two distinct alpha6beta2(*) nAChR subtypes. These include a very high (femtomolar) and a high (picomolar) affinity site, with approximately 40% of the sites in the very high affinity form. It is noteworthy that only the high affinity form was detected in alpha4 nAChR-null mutant mice. Because (125)I-alpha CtxMII binds primarily to alpha6alpha4beta2beta3 and alpha6beta2beta3 nAChR subtypes in mouse striatum, these data suggest that the population lost in the alpha4 knockout mice was the alpha6alpha4beta2beta3 subtype. We next investigated the effect of nigrostriatal lesioning on these two striatal alpha6beta2(*) populations in two animal models and in Parkinson's disease. There was a preferential loss of the very high affinity subtype in striatum of mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), monkeys treated with MPTP, and patients with Parkinson's disease. These data suggest that dopaminergic terminals expressing the alpha6alpha4beta2beta3 population are selectively vulnerable to nigrostriatal damage. This latter nAChR subtype, identified with alpha-CtxMII E11A, may therefore provide a unique marker for dopaminergic terminals particularly sensitive to nigrostriatal degeneration in Parkinson's disease. PMID- 17409285 TI - Trapping of a nonpeptide ligand by the extracellular domains of the gonadotropin releasing hormone receptor results in insurmountable antagonism. AB - Drugs that exhibit insurmountable antagonism are proposed to provide improved clinical efficacy through extended receptor blockade. Long-term suppression of the gonadotropin-releasing hormone receptor (GnRHR) is an important therapeutic approach for a number of sex hormone-dependent diseases. In this study, we describe the mechanism and structural components required for insurmountable activity of a GnRHR antagonist. TAK-013 behaves as an insurmountable antagonist at the human receptor (hGnRHR) but as a surmountable antagonist at the macaque receptor (mGnRHR). Mutation of the eight residues that differ between hGnRHR and mGnRHR identified Ser-203 and Leu-300 in extracellular loops (ECL) 2 and 3 of hGnRHR as essential for the insurmountability of TAK-013. Substitution of the corresponding residues in mGnRHR with Ser and Leu (mGnRHR-P203S/V300L) converts TAK-013 to an insurmountable antagonist. In addition, mutation of Met-24 to Leu in the amino terminus of hGnRHR also ablates the insurmountable antagonism of TAK 013. The mechanism of insurmountability of TAK-013 was determined to be governed by its rate of dissociation from the receptor. Although the association rates of TAK-013 to hGnRHR, mGnRHR, and mGnRHR-P203S/V300L do not differ, the dissociation rate half-life correlates closely with the degree of insurmountability observed (169, 9, and 55 min, respectively). Taken together, these data suggest a model of the GnRHR in which ECL2, ECL3, and the amino terminus engage with TAK-013 upon its binding to the transmembrane region of the receptor. These additional interactions form a "trap door" above TAK-013, restricting its dissociation and thus resulting in its insurmountability. PMID- 17409286 TI - Notch signaling in development and cancer. AB - Notch is an evolutionarily conserved local cell signaling mechanism that participates in a variety of cellular processes: cell fate specification, differentiation, proliferation, apoptosis, adhesion, epithelial-mesenchymal transition, migration, and angiogenesis. These processes can be subverted in Notch-mediated pathological situations. In the first part of this review, we will discuss the role of Notch in vertebrate central nervous system development, somitogenesis, cardiovascular and endocrine development, with attention to the mechanisms by which Notch regulates cell fate specification and patterning in these tissues. In the second part, we will review the molecular aspects of Notch mediated neoplasias, where Notch can act as an oncogene or as a tumor suppressor. From all these studies, it becomes evident that the outcome of Notch signaling is strictly context-dependent and differences in the strength, timing, cell type, and context of the signal may affect the final outcome. It is essential to understand how Notch integrates inputs from other signaling pathways and how specificity is achieved, because this knowledge may be relevant for future therapeutic applications. PMID- 17409287 TI - Tumor cell-organ microenvironment interactions in the pathogenesis of cancer metastasis. AB - The process of cancer metastasis is sequential and selective and contains stochastic elements. The growth of metastases represents the endpoint of many lethal events that few tumor cells can survive. Primary tumors consist of multiple subpopulations of cells with heterogeneous metastatic properties, and the outcome of metastasis depends on the interplay of tumor cells with various host factors. The findings that different metastases can originate from different progenitor cells account for the biological diversity that exists among various metastases. Even within a solitary metastasis of proven clonal origin, however, heterogeneity of biological characteristics can develop rapidly. The pathogenesis of metastasis depends on multiple interactions of metastatic cells with favorable host homeostatic mechanisms. Interruption of one or more of these interactions can lead to the inhibition or eradication of cancer metastasis. For many years, all of our efforts to treat cancer have concentrated on the inhibition or destruction of tumor cells. Strategies both to treat tumor cells (such as chemotherapy and immunotherapy) and to modulate the host microenvironment (including the tumor vasculature) should offer additional approaches for cancer treatment. The recent advances in our understanding of the biological basis of cancer metastasis present unprecedented possibilities for translating basic research to the clinical reality of cancer treatment. PMID- 17409288 TI - The role of alpha-cell dysregulation in fasting and postprandial hyperglycemia in type 2 diabetes and therapeutic implications. AB - The hyperglycemic activity of pancreatic extracts was encountered some 80 yr ago during efforts to optimize methods for the purification of insulin. The hyperglycemic substance was named "glucagon," and it was subsequently determined that glucagon is a 29-amino acid peptide synthesized and released from pancreatic alpha-cells. This article begins with a brief overview of the discovery of glucagon and the contributions that somatostatin and a sensitive and selective assay for pancreatic (vs. gut) glucagon made to understanding the physiological and pathophysiological roles of glucagon. Studies utilizing these tools to establish the function of glucagon in normal nutrient homeostasis and to document a relative glucagon excess in type 2 diabetes mellitus (T2DM) and precursors thereof are then discussed. The evidence that glucagon excess contributes to the development and maintenance of fasting hyperglycemia and that failure to suppress glucagon secretion contributes to postprandial hyperglycemia is then reviewed. Although key human studies are emphasized, salient animal studies highlighting the importance of glucagon in normal and defective glucoregulation are also described. The past eight decades of research in this area have led to development of new therapeutic approaches to treating T2DM that have been shown to, or are expected to, improve glycemic control in patients with T2DM in part by improving alpha-cell function or by blocking glucagon action. Accordingly, this review ends with a discussion of the status and therapeutic potential of glucagon receptor antagonists, alpha-cell selective somatostatin agonists, glucagon-like peptide-1 agonists, and dipeptidyl peptidase-IV inhibitors. Our overall conclusions are that there is considerable evidence that relative hyperglucagonemia contributes to fasting and postprandial hyperglycemia in patients with T2DM, and there are several new and emerging pharmacotherapies that may improve glycemic control in part by ameliorating the hyperglycemic effects of this relative glucagon excess. PMID- 17409289 TI - Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law. PMID- 17409290 TI - Counterpoint: airway smooth muscle is not useful. PMID- 17409293 TI - Comments on point: counterpoint: "airway smooth muscle is/is not useful". PMID- 17409294 TI - Comments on point: counterpoint: "the muscle metaboreflex does/does not restore blood flow to contracting muscles". PMID- 17409295 TI - Commentary on viewpoint "Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law". PMID- 17409296 TI - Commentary on viewpoint "Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law". PMID- 17409297 TI - Commentary on viewpoint "Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law". PMID- 17409298 TI - Commentary on viewpoint "Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law". PMID- 17409299 TI - Commentary on viewpoint "Heliox, nitrox, and trimix diving; hyperbaric oxygen treatment; and a flaw in Henry's law. PMID- 17409301 TI - Short-acting NO donor and decompression sickness in humans. PMID- 17409303 TI - Are neuroepithelial bodies a part of pulmonary slowly adapting receptors? PMID- 17409305 TI - Oxidative stress causes renal dopamine D1 receptor dysfunction and hypertension via mechanisms that involve nuclear factor-kappaB and protein kinase C. AB - Renal dopamine, via activation of D1 receptors, plays a role in maintaining sodium homeostasis and BP. There exists a defect in renal D1 receptor function in hypertension, diabetes, and aging, conditions that are associated with oxidative stress. However, the exact underlying mechanism of the oxidative stress-mediated impaired D1 receptor signaling and hypertension is not known. The effect of oxidative stress on renal D1 receptor function was investigated in healthy animals. Male Sprague-Dawley rats received tap water (vehicle) and 30 mM L buthionine sulfoximine (BSO), an oxidant, with and without 1 mM tempol for 2 wk. Compared with vehicle, BSO treatment caused oxidative stress and increase in BP, which was accompanied by defective D1 receptor G-protein coupling and loss of natriuretic response to SKF38393. BSO treatment also increased NF-kappaB nuclear translocation, protein kinase C (PKC) activity and expression, G-protein-coupled receptor kinase-2 (GRK-2) membranous translocation, and D1 receptor serine phosphorylation. In BSO-treated rats' supplementation of tempol decreased oxidative stress, normalized BP, and restored D1 receptor G-protein coupling and natriuretic response to SKF38393. Tempol also normalized NF-kappaB translocation, PKC activity and expression, GRK-2 sequestration, and D1 receptor serine phosphorylation. In conclusion, these results show that oxidative stress activates NF-kappaB, causing an increase in PKC activity, which leads to GRK-2 translocation and subsequent D1 receptor hyper-serine phosphorylation and uncoupling. The functional consequence of this phenomenon was the inability of SKF38393 to inhibit Na/K-ATPase activity and promote sodium excretion, which may have contributed to increase in BP. Tempol reduced oxidative stress and thereby restored D1 receptor function and normalized BP. PMID- 17409306 TI - Progressive exercise for anabolism in kidney disease (PEAK): a randomized, controlled trial of resistance training during hemodialysis. AB - Skeletal muscle wasting is common and insidious in patients who receive maintenance hemodialysis treatment for the management of ESRD. The objective of this study was to determine whether 12 wk of high-intensity, progressive resistance training (PRT) administered during routine hemodialysis treatment could improve skeletal muscle quantity and quality versus usual care. Forty-nine patients (62.6 +/- 14.2 yr; 0.3 to 16.7 yr on dialysis) were recruited from the outpatient hemodialysis unit of the St. George Public Hospital (Sydney, Australia). Patients were randomized to PRT + usual care (n = 24) or usual care control only (n = 25). The PRT group performed two sets of 10 exercises at a high intensity (15 to 17/20 on the Borg Scale) using free weights, three times per week for 12 wk during routine hemodialysis treatment. Primary outcomes included thigh muscle quantity (cross-sectional area [CSA]) and quality (intramuscular lipid content via attenuation) evaluated by computed tomography scan. Secondary outcomes included muscle strength, exercise capacity, body circumference measures, proinflammatory cytokine C-reactive protein, and quality of life. There was no statistically significant difference in muscle CSA change between groups. However, there were statistically significant improvements in muscle attenuation, muscle strength, mid-thigh and mid-arm circumference, body weight, and C-reactive protein in the PRT group relative to the nonexercising control group. These findings suggest that patients with ESRD can improve skeletal muscle quality and derive other health-related adaptations solely by engaging in a 12-wk high intensity PRT regimen during routine hemodialysis treatment sessions. Longer training durations or more sensitive analysis techniques may be required to document alterations in muscle CSA. PMID- 17409307 TI - Diurnal blood pressure changes one year after kidney transplantation: relationship to allograft function, histology, and resistive index. AB - Loss of circadian BP change has been linked to target organ damage and accelerated kidney function loss in hypertensive patients with and without chronic kidney disease. Ambulatory BP-derived data from 119 consecutive kidney transplant recipients who presented for the first annual evaluation were examined in relation to allograft function, histology, and ultrasound findings. A total of 101 (85%) patients were receiving antihypertensive medications (median 2), and 85 (71%) achieved target awake average systolic BP (SBP) of <135 mmHg. A day-night change in SBP by 10% or more (dippers) was detected in 29 (24%). Dipping status was associated with younger recipient age, lack of diabetes, low chronic vascular score, and low resistive index. Nondippers and reverse dippers had lower GFR compared with dippers (P = 0.04). For every 10% nocturnal drop in SBP, GFR increased by 4.6 ml/min per 1.73 m(2) (R = 0.3, P = 0.003). Nondippers and reverse dippers were equally common in recipients with normal histology and in those with pathologic findings on surveillance biopsy. On multivariate analysis, percentage of nocturnal fall in SBP and elevated resistive index independently correlated with GFR. This study indicates that lack of nocturnal fall in SBP is related to poor allograft function, high chronic vascular score, and high resistive index irrespective of allograft fibrosis. Further studies are needed to determine whether restoration of normal BP pattern will confer better allograft outcome. PMID- 17409308 TI - Weekly enzyme replacement therapy may slow decline of renal function in patients with Fabry disease who are on long-term biweekly dosing. AB - This study was performed to determine whether adult male patients with Fabry disease who demonstrate a continuing decline in renal function despite 2 to 4 yr of conventionally dosed agalsidase alfa therapy (0.2 mg/kg every other week [EOW]) show an improved slope of decline with weekly administration using the same dosage. Eleven (27%) of 41 adult male patients with Fabry disease who participated in long-term agalsidase alfa clinical trials and who had demonstrated a slope of decline in estimated GFR (eGFR) of > or =5 ml/min per 1.73 m(2)/yr while receiving long-term treatment with agalsidase alfa at the currently recommended dosage of 0.2 mg/kg, infused EOW, were enrolled in this open-label, prospective study. Patients were switched from EOW to weekly infusions and followed for an additional 24 mo. Before switching to weekly dosing, eGFR was 53.7 +/- 6.3 ml/min per 1.73 m(2) (mean +/- SEM), and mean rate of change in eGFR was -8.0 +/- 0.8 ml/min per 1.73 m(2)/yr. During the 24-mo follow-up period after switching to weekly dosing, the mean rate of change in eGFR was observed to slow to -3.3 +/- 1.4 ml/min/1.73 m(2)/yr (P = 0.01 versus EOW). After switching to weekly dosing, three patients demonstrated an improvement in eGFR and six patients demonstrated a slowing in the rate of eGFR decline; only two patients failed to improve their eGFR slope. A multiple regression model confirmed that the weekly infusion regimen was the strongest explanatory variable for the change in eGFR (P = 0.0008), with a weaker contribution from the concomitant use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers (P = 0.02). These results suggest that weekly infusions of agalsidase alfa at a dosage of 0.2 mg/kg may be beneficial in the subgroup of patients who have Fabry disease and whose kidney function continues to decline after 2 to 4 yr or more of standard EOW dosing. PMID- 17409309 TI - High NPHP1 and NPHP6 mutation rate in patients with Joubert syndrome and nephronophthisis: potential epistatic effect of NPHP6 and AHI1 mutations in patients with NPHP1 mutations. AB - Joubert syndrome (JS) is an autosomal recessive disorder that is described in patients with cerebellar ataxia, mental retardation, hypotonia, and neonatal respiratory dysregulation. Kidney involvement (nephronophthisis or cystic renal dysplasia) is associated with JS in one fourth of known cases. Mutations in three genes--AHI1, NPHP1, and NPHP6--have been identified in patients with JS. However, because NPHP1 mutations usually cause isolated nephronophthisis, the factors that predispose to the development of neurologic involvement are poorly understood. In an attempt to identify such genetic determinants, a cohort of 28 families with nephronophthisis and at least one JS-related neurologic symptom were screened for mutations in AHI1, NPHP1, and NPHP6 genes. NPHP1 and NPHP6 homozygous or compound heterozygous mutations were found in 13 (46%) and six (21%) unrelated patients, respectively. Two of the 13 patients with NPHP1 mutations carried either a heterozygous truncating mutation in NPHP6 or a heterozygous missense mutation in AHI1. Furthermore, five patients with NPHP1 mutations carried the AHI1 variant R830W, which was predicted to be "possibly damaging" and was found with significantly higher frequency than in healthy control subjects and in patients with NPHP1 mutations without neurologic symptoms (five of 26 versus four of 276 and three of 152 alleles; P < 0.001 and P < 0.002, respectively). In contrast to the variable neurologic and milder retinal phenotype of patients with NPHP1 mutations, patients with NPHP6 mutations presented with a more severe neurologic and retinal phenotype. In conclusion, NPHP1 and NPHP6 are major genes of nephronophthisis associated with JS. Epistatic effects that are provided by heterozygous NPHP6 and AHI1 mutations and variants may contribute to the appearance of extrarenal symptoms in patients with NPHP1 mutations. PMID- 17409310 TI - Distal renal tubular acidosis in mice lacking the AE1 (band3) Cl-/HCO3- exchanger (slc4a1). AB - Mutations in the human gene that encodes the AE1 Cl(-)/HCO(3)(-) exchanger (SLC4A1) cause autosomal recessive and dominant forms of distal renal tubular acidosis (dRTA). A mouse model that lacks AE1/slc4a1 (slc4a1-/-) exhibited dRTA characterized by spontaneous hyperchloremic metabolic acidosis with low net acid excretion and, inappropriately, alkaline urine without bicarbonaturia. Basolateral Cl(-)/HCO(3)(-) exchange activity in acid-secretory intercalated cells of isolated superfused slc4a1-/- medullary collecting duct was reduced, but alternate bicarbonate transport pathways were upregulated. Homozygous mice had nephrocalcinosis associated with hypercalciuria, hyperphosphaturia, and hypocitraturia. A severe urinary concentration defect in slc4a1-/- mice was accompanied by dysregulated expression and localization of the aquaporin-2 water channel. Mice that were heterozygous for the AE1-deficient allele had no apparent defect. Thus, the slc4a1-/- mouse is the first genetic model of complete dRTA and demonstrates that the AE1/slc4a1 Cl(-)/HCO(3)(-) exchanger is required for maintenance of normal acid-base homeostasis by distal renal regeneration of bicarbonate in the mouse as well as in humans. PMID- 17409311 TI - Sodium-hydrogen exchanger regulatory factor-1 interacts with mouse urate transporter 1 to regulate renal proximal tubule uric acid transport. AB - Sodium-hydrogen exchanger regulatory factor-1-deficient (NHERF-1(-/-)) mice demonstrate increases in the urinary excretion of phosphate, calcium, and uric acid associated with interstitial deposition of calcium in the papilla of the kidney. These studies examine the role of NHERF-1 in the tubular reabsorption of uric acid and regulation of mouse urate transporter 1 (mURAT1), a newly described transporter that is responsible for the renal tubular reabsorption of uric acid. In primary cultures of mouse renal proximal tubule cells, uric acid uptake was significantly lower in NHERF-1(-/-) cells compared with wild-type cells over a large range of uric acid concentrations in the media. Western immunoblotting revealed a 56 +/- 6% decrease in the brush border membrane (BBM) expression of mURAT1 in NHERF-1(-/-) compared with wild-type control kidneys (P < 0.05). Confocal microscopy confirmed the reduced apical membrane expression of mURAT1 in NHERF-1(-/-) kidneys and demonstrated mislocalization of mURAT1 to intracellular vesicular structures. Para-aminohippurate significantly inhibited uric acid uptake in wild-type cells (41 +/- 2%) compared with NHERF-1(-/-) cells (8.2 +/- 3%). Infection of NHERF-1(-/-) cells with adenovirus-green fluorescence protein NHERF-1 resulted in significantly higher rates of uric acid transport (15.4 +/- 1.1 pmol/microg protein per 30 min) compared with null cells that were infected with control adenovirus-green fluorescence protein (7.9 +/- 0.3) and restoration of the inhibitory effect of para-aminohippurate (% inhibition 34 +/- 4%). These findings indicate that NHERF-1 exerts a significant effect on the renal tubular reabsorption of uric acid in the mouse by modulating the BBM abundance of mURAT1 and possibly other BBM uric acid transporters. PMID- 17409312 TI - Sustained, long-term renal stabilization after 54 months of agalsidase beta therapy in patients with Fabry disease. AB - Fabry disease, an inherited deficiency of the lysosomal enzyme alpha galactosidase A, causes progressive intralysosomal accumulation of globotriaosylceramide (GL-3) and premature death from renal, cardiac, and cerebrovascular manifestations. To determine the long-term safety and efficacy of recombinant human alpha-galactosidase A, an open-label, phase III extension study was conducted, involving 58 patients who had classic Fabry disease and completed a 20-wk, double-blind, randomized, placebo-controlled, phase III study of agalsidase beta and were transitioned to an extension trial to receive biweekly 1 mg/kg agalsidase beta for up to an additional 54 mo. GL-3 accumulation was evaluated in the capillary endothelia of the skin, kidney, and heart. Renal function was assessed. By month 54, all patients with optional kidney biopsies (n = 8) maintained complete GL-3 clearance in renal capillary endothelial cells and multiple cell types. Continued, complete clearance of skin (31 of 36) and heart (six of eight) capillary endothelium was demonstrated. Mean plasma GL-3 levels remained decreased in the normal range. Median serum creatinine and estimated GFR remained stable (normal) in patients with renal data at month 54 (n = 41). Six patients had renal disease progression; most (four of six) were older than 40 yr and had significant proteinuria at baseline and evidence of sclerotic glomeruli pretreatment. Adverse events were generally mild and unrelated to treatment. The most common treatment-related adverse events were infusion-associated reactions, which decreased over time. Long-term agalsidase beta therapy stabilizes renal function in patients without renal involvement at baseline, maintains reduction of plasma GL-3, and sustains GL-3 clearance in capillary endothelial cells and multiple renal cell types. PMID- 17409313 TI - Uric acid heralds ischemic tissue injury to mobilize endothelial progenitor cells. AB - Understanding the nature of endogenous mechanisms for mobilization of stem/progenitor cells is predicated on the identification of injury-induced substances that are released from a damaged organ and capable of producing a distant effect. Although different substances that mobilize endothelial progenitor cells (EPCs) have been proposed, their potential to signal injury and afford postischemic renoprotection and repair remains obscure. Uric acid (UA) is consistently overproduced by ischemic tissues and has been shown to exert immunomodulatory functions. It was hypothesized that UA and/or its precursors might serve as injury signals that are capable of mobilizing EPCs in acute renal ischemia. Indeed, FVB/NJ mice that were subjected to acute renal ischemia showed a transient surge in UA level in the peripheral blood. Single-dose treatment with UA, as well as acute hyperuricemia induced by the inhibition of uricase, caused a robust mobilization of EPCs, whereas administration of adenosine or inosine seemed to lack this effect. Moreover, pretreatment of mice with a single dose of UA afforded significant renoprotection against ischemic injury. In animals with chronic hyperuricemia (induced by continuous 2-wk treatment with a uricase inhibitor oxonic acid), EPC mobilization was blunted and renoprotective effects were absent. In conclusion, acute elevation of UA acts as "physiologic," fast acting endogenous mediator of EPC mobilization and renoprotection, consistent with its novel function in pharmacologic preconditioning. Both of these actions are lacking in mice with chronic hyperuricemia. In summary, a transient surge in UA concentration may serve as a universal herald of tissue injury to accelerate the recruitment of EPCs. PMID- 17409314 TI - Dimethylarginine dimethylaminohydrolase prevents progression of renal dysfunction by inhibiting loss of peritubular capillaries and tubulointerstitial fibrosis in a rat model of chronic kidney disease. AB - Asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, is mainly degraded by dimethylarginine dimethylaminohydrolase (DDAH). It was recently reported that reduced DDAH expression could contribute to ADMA accumulation and subsequent elevation of BP in an experimental model of chronic kidney disease (CKD). ADMA is a strong predictor of the progression of CKD as well. However, a role for the ADMA-DDAH in the pathogenesis of CKD remains to be elucidated. This study investigated the effects of DDAH-elicited ADMA lowering on renal function and pathology in a rat remnant kidney model. Four weeks after five sixths subtotal nephrectomy (Nx), the rats were given tail-vein injections of recombinant adenovirus vector encoding DDAH-I (Adv-DDAH) or control vector expressing bacterial beta-galactosidase (Adv-LZ) or orally administered 20 mg/kg per d hydralazine (Hyz), which served as a BP control model. In comparison with Adv-LZ or Hyz administration, Adv-DDAH decreased plasma levels of ADMA and inhibited the deterioration of renal dysfunction. Plasma levels of ADMA were associated with decreased number of peritubular capillaries, increased tubulointerstitial fibrosis, and proteinuria levels in Nx rats. These changes were progressed in Adv-LZ-or Hyz-treated Nx rats, which were ameliorated by DDAH overexpression. In addition, semiquantitative reverse transcriptase-PCR and immunohistochemistry for TGF-beta revealed that Adv-DDAH inhibited upregulation of TGF-beta expression in Nx rats. These data suggest that ADMA may be involved in peritubular capillary loss and tubulointerstitial fibrosis, thereby contributing to the progression of CKD. Substitution of DDAH protein or enhancement of its activity may become a novel therapeutic strategy for the treatment of CKD. PMID- 17409315 TI - Neutralization of macrophage-stimulating protein ameliorates renal injury in anti thy 1 glomerulonephritis. AB - Macrophage-stimulating protein (MSP) is a scatter factor that causes cell proliferation and migration, and receptor origin nantaise (RON) is its receptor. RON is expressed in macrophages and mesangial cells, and MSP is produced by renal tubular cells. This study investigated whether MSP/RON participate in the pathogenesis of anti-Thy 1 nephritis, a glomerular disease that is characterized by invasion of circulating monocytes into glomeruli and migration and proliferation of mesangial cells. In vivo, renal function and histopathology were studied in rats that had anti-Thy 1 disease and were untreated and treated with a neutralizing anti-MSP antibody. In vitro, whether monocytes express RON and whether MSP has a chemotactic effect on monocytes were studied. In vivo, in anti Thy 1 disease, MSP was expressed de novo in glomeruli, and neutralization of MSP attenuated the rise in serum creatinine and proteinuria, stopped glomerular neutrophil and monocyte influx, protected from glomerular injury, and lessened mesangial cell overgrowth. In vitro, unstimulated monocytes did not express RON, but the stimulation with LPS induced de novo RON expression. LPS-stimulated monocytes were attracted by MSP. These results demonstrate a pathogenic role of the MSP/RON system in anti-Thy 1 nephritis. PMID- 17409316 TI - Urinary angiotensinogen as a marker of intrarenal angiotensin II activity associated with deterioration of renal function in patients with chronic kidney disease. AB - In chronic kidney disease (CKD), enhanced intrarenal angiotensin II (AngII) is involved in deterioration of renal function, but it is difficult to measure it. For assessment of the potential of urinary angiotensinogen as a marker of intrarenal AngII activity, the correlation of plasma and urinary renin angiotensin system components, including angiotensinogen, with deterioration of renal function was investigated in 80 patients who had CKD and were not treated with AngII blocking agents. Changes that were induced by 14 d of losartan treatment (25 mg/d) were also measured in 28 patients. Angiotensinogen was measured by RIA of AngI after incubation with renin. Urinary angiotensinogen levels were greater in patients with low estimated GFR and elevated urinary protein and type IV collagen and correlated with renal AngII and type I collagen immunostaining intensities. The risk for deterioration of renal function (i.e., estimated GFR decline of >2.5 ml/min per yr) during a mean follow-up period of 23 mo (maximum 43 mo) was associated with urinary angiotensinogen of >3.0 nmol AngI equivalent per 1 g of urinary creatinine (AngI Eq/g Cre) at enrollment (hazard ratio 3.52). The event-free survival for deterioration of renal function was better in patients with urinary angiotensinogen <3.0 nmol AngI Eq/g Cre than those >3.0 nmol AngI Eq/g Cre. Losartan reduced urinary and plasma angiotensinogen, urinary protein and type IV collagen, and systolic BP, despite concomitant increases in plasma renin and AngII. These data suggest that urinary angiotensinogen is a potentially suitable marker of intrarenal AngII activity associated with increased risk for deterioration of renal function in patients with CKD. PMID- 17409317 TI - Albuminuria is a target for renoprotective therapy independent from blood pressure in patients with type 2 diabetic nephropathy: post hoc analysis from the Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) trial. AB - Albuminuria reduction could be renoprotective in hypertensive patients with diabetic nephropathy. However, the current use of renin-angiotensin-system intervention is targeted to BP only. Therefore, this study investigated the adequacy of this approach in 1428 patients with hypertension and diabetic nephropathy from the placebo-controlled Reduction of Endpoints in NIDDM with the Angiotensin II Antagonist Losartan (RENAAL) study. Investigated were the extent of discordance in treatment effects on systolic BP (SBP) and albuminuria and its association with renal outcome in a multivariate Cox model. Among patients with a reduced SBP during treatment, a lack of albuminuria reduction was observed in 37, 26, and 51% (total, losartan, and placebo, respectively) at month 6. SBP or albuminuria reduction was associated with a lower risk for ESRD, whereas combined SBP and albuminuria reduction was associated with the lowest risk for events. Across all categories of SBP change, a progressively lower ESRD hazard ratio was observed with a larger albuminuria reduction. A lower residual level of albuminuria was also associated with lower ESRD risk. In conclusion, changes in albuminuria are not concordant in a substantial proportion of patients when titrated for BP. Meanwhile, the ESRD risk showed a clear dependence on albuminuria reduction. The ESRD risk also showed dependence on the residual level of albuminuria, even in patients who reached the current SBP target. Antihypertensive treatment that is aimed at improving renal outcomes in patients with diabetic nephropathy may therefore require a dual strategy, targeting both SBP and albuminuria reduction. PMID- 17409318 TI - Competition between 24:5n-3 and ALA for Delta 6 desaturase may limit the accumulation of DHA in HepG2 cell membranes. AB - The use of Delta 6 desaturase (D6D) twice in the conversion of alpha-linolenic acid (ALA; 18:3n-3) to docosahexaenoic acid (DHA; 22:6n-3) suggests that this enzyme may play a key regulatory role in the synthesis and accumulation of DHA from ALA. We examined this using an in vitro model of fatty acid metabolism to measure the accumulation of the long-chain metabolites of ALA in HepG2 cell phospholipids. The accumulation of ALA, eicosapentaenoic acid (20:5n-3), docosapentaenoic acid (22:5n-3), and 24:5n-3 in cell phospholipids was linearly related to the concentration of supplemented ALA over the range tested (1.8-72 microM). The accumulation of the post-D6D products of 22:5n-3, 24:6n-3 and DHA, in cell phospholipids was saturated at concentrations of >18 microM ALA. Supplementation of HepG2 cells with preformed DHA revealed that, although the accumulation of DHA in cell phospholipids approached saturation, the level of DHA in cell phospholipids was significantly greater compared with the accumulation of DHA from ALA, indicating that the accumulation of DHA from ALA was not limited by incorporation. The parallel pattern of accumulation of 24:6n-3 and DHA in response to increasing concentrations of ALA suggests that the competition between 24:5n-3 and ALA for D6D may contribute to the limited accumulation of DHA in cell membranes. PMID- 17409319 TI - Benefits with risks--Bush's tax-based health care proposals. PMID- 17409320 TI - Making motherhood safe in developing countries. PMID- 17409321 TI - Influence of computer-aided detection on performance of screening mammography. AB - BACKGROUND: Computer-aided detection identifies suspicious findings on mammograms to assist radiologists. Since the Food and Drug Administration approved the technology in 1998, it has been disseminated into practice, but its effect on the accuracy of interpretation is unclear. METHODS: We determined the association between the use of computer-aided detection at mammography facilities and the performance of screening mammography from 1998 through 2002 at 43 facilities in three states. We had complete data for 222,135 women (a total of 429,345 mammograms), including 2351 women who received a diagnosis of breast cancer within 1 year after screening. We calculated the specificity, sensitivity, and positive predictive value of screening mammography with and without computer aided detection, as well as the rates of biopsy and breast-cancer detection and the overall accuracy, measured as the area under the receiver-operating characteristic (ROC) curve. RESULTS: Seven facilities (16%) implemented computer aided detection during the study period. Diagnostic specificity decreased from 90.2% before implementation to 87.2% after implementation (P<0.001), the positive predictive value decreased from 4.1% to 3.2% (P=0.01), and the rate of biopsy increased by 19.7% (P<0.001). The increase in sensitivity from 80.4% before implementation of computer-aided detection to 84.0% after implementation was not significant (P=0.32). The change in the cancer-detection rate (including invasive breast cancers and ductal carcinomas in situ) was not significant (4.15 cases per 1000 screening mammograms before implementation and 4.20 cases after implementation, P=0.90). Analyses of data from all 43 facilities showed that the use of computer-aided detection was associated with significantly lower overall accuracy than was nonuse (area under the ROC curve, 0.871 vs. 0.919; P=0.005). CONCLUSIONS: The use of computer-aided detection is associated with reduced accuracy of interpretation of screening mammograms. The increased rate of biopsy with the use of computer-aided detection is not clearly associated with improved detection of invasive breast cancer. PMID- 17409322 TI - Invariant natural killer T cells in asthma and chronic obstructive pulmonary disease. AB - BACKGROUND: The number of type 2 helper CD4+ T cells is increased in the airways of persons with asthma. Whether the majority of these cells are class II major histocompatibility-complex-restricted cells or are among the recently identified CD1d-restricted invariant natural killer T cells is a matter of controversy. We studied the frequency of invariant natural killer T cells in the airways of subjects with mild or moderately severe asthma to investigate the possibility of an association between the number of invariant natural killer T cells in the airway and disease severity. We also studied whether an increased number of these cells is a feature of chronic obstructive pulmonary disease (COPD). METHODS: We enumerated invariant natural killer T cells by flow cytometry with the use of CD1d tetramers loaded with alpha-galactosylceramide and antibodies specific to the invariant natural killer T-cell receptor in samples of bronchoalveolar-lavage fluid, induced sputum, and bronchial-biopsy specimens obtained from subjects with mild or moderately severe asthma, subjects with COPD, and healthy control subjects. Real-time polymerase-chain-reaction analysis was performed on bronchoalveolar-lavage cells for evidence of gene expression of the invariant natural killer T-cell receptor. RESULTS: Fewer than 2% of the T cells obtained from all subjects on airway biopsy, bronchoalveolar lavage, and sputum induction were invariant natural killer T cells, with no significant differences among the three groups of subjects. No expression of messenger RNA for the invariant natural killer T-cell-receptor domains Valpha24 and Vbeta11 was detected in bronchoalveolar-lavage cells from subjects with asthma. CONCLUSIONS: Invariant natural killer T cells are found in low numbers in the airways of subjects with asthma, subjects with COPD, and controls. PMID- 17409323 TI - Vitamins and perinatal outcomes among HIV-negative women in Tanzania. AB - BACKGROUND: Prematurity and low birth weight are associated with high perinatal and infant mortality, especially in developing countries. Maternal micronutrient deficiencies may contribute to these adverse outcomes. METHODS: In a double-blind trial in Dar es Salaam, Tanzania, we randomly assigned 8468 pregnant women (gestational age of fetus, 12 to 27 weeks) who were negative for human immunodeficiency virus infection to receive daily multivitamins (including multiples of the recommended dietary allowance) or placebo. All the women received prenatal supplemental iron and folic acid. The primary outcomes were low birth weight (<2500 g), prematurity, and fetal death. RESULTS: The incidence of low birth weight was 7.8% among the infants in the multivitamin group and 9.4% among those in the placebo group (relative risk, 0.82; 95% confidence interval [CI], 0.70 to 0.95; P=0.01). The mean difference in birth weight between the groups was modest (67 g, P<0.001). The rates of prematurity were 16.9% in the multivitamin group and 16.7% in the placebo group (relative risk, 1.01; 95% CI, 0.91 to 1.11; P=0.87), and the rates of fetal death were 4.3% and 5.0%, respectively (relative risk, 0.87; 95% CI, 0.72 to 1.05; P=0.15). Supplementation reduced both the risk of a birth size that was small for gestational age (<10th percentile; 10.7% in the multivitamin group vs. 13.6% in the placebo group; relative risk, 0.77; 95% CI, 0.68 to 0.87; P<0.001) and the risk of maternal anemia (hemoglobin level, <11 g per deciliter; relative risk, 0.88; 95% CI, 0.80 to 0.97; P=0.01), although the difference in the mean hemoglobin levels between the groups was small (0.2 g per deciliter, P<0.001). CONCLUSIONS: Multivitamin supplementation reduced the incidence of low birth weight and small-for gestational-age births but had no significant effects on prematurity or fetal death. Multivitamins should be considered for all pregnant women in developing countries. (ClinicalTrials.gov number, NCT00197548 [ClinicalTrials.gov].). PMID- 17409324 TI - Mutations in VANGL1 associated with neural-tube defects. AB - Neural-tube defects such as anencephaly and spina bifida constitute a group of common congenital malformations caused by complex genetic and environmental factors. We have identified three mutations in the VANGL1 gene in patients with familial types (V239I and R274Q) and a sporadic type (M328T) of the disease, including a spontaneous mutation (V239I) appearing in a familial setting. In a protein-protein interaction assay V239I abolished interaction of VANGL1 protein with its binding partners, disheveled-1, -2, and -3. These findings implicate VANGL1 as a risk factor in human neural-tube defects. PMID- 17409325 TI - Clinical practice. Prophylaxis for thromboembolism in hospitalized medical patients. PMID- 17409327 TI - Images in clinical medicine. A medical mystery--skin discolorations in a bank employee. PMID- 17409326 TI - Viral hepatitis in HIV infection. PMID- 17409328 TI - Clinical problem-solving. Building a diagnosis from the ground up--a 49-year-old man came to the clinic with a 1-week history of suprapubic pain and fever. PMID- 17409329 TI - Breast imaging and computer-aided detection. PMID- 17409330 TI - Natural killer T cells in asthma--toward increased understanding. PMID- 17409331 TI - Diagnosis of ventilator-associated pneumonia. PMID- 17409332 TI - Lapatinib plus capecitabine in breast cancer. PMID- 17409333 TI - Biomarkers for prediction of cardiovascular events. PMID- 17409334 TI - Door-to-balloon time in acute myocardial infarction. PMID- 17409335 TI - Whipple's disease. PMID- 17409336 TI - Multiple-triazole-resistant aspergillosis. PMID- 17409337 TI - Retraction: Hussain HM, Hotopf M, Oyebode F. Atypical antipsychotic drugs and Alzheimer's disease. N Engl J Med 2007;356:416. PMID- 17409338 TI - Approach to the growth hormone-deficient child during transition to adulthood. AB - The observation that some adults with childhood-onset GH deficiency have low bone mineral density, low lean body mass, diminished quality of life, abnormal lipids, and impaired cardiac function, all of which may improve after treatment with GH, has prompted pediatric endocrinologists to reevaluate the practice of discontinuing GH in all patients after attainment of final adult height. The treatment of adolescents to prevent the metabolic complications of GH deficiency is an emerging practice. Studies addressing the evaluation and care of adolescents during this period and the benefits of GH in this setting are conflicting. Our approach in determining which adolescents to retest, when and how to test for persistent GH deficiency, and which subjects to treat is discussed in the context of available clinical data. PMID- 17409339 TI - Transient hypophysitis after cytotoxic T lymphocyte-associated antigen 4 (CTLA4) blockade. PMID- 17409340 TI - Clinical translation of "a diabetes outcome progression trial": ADOPT appropriate combination oral therapies in type 2 diabetes. PMID- 17409341 TI - Of the fit and the fat: mitochondrial abnormalities and type 2 diabetes mellitus. PMID- 17409342 TI - Thiazolidinediones: new evidence of bone loss. PMID- 17409343 TI - Ruling out a suspect: the role of beta-catenin mutation in benign parathyroid neoplasia. PMID- 17409344 TI - Vitamin K supplementation during oral anticoagulation: concerns. PMID- 17409346 TI - Acute thrombocytopenia caused by sensitivity to the glucuronide conjugate of acetaminophen. PMID- 17409347 TI - Imatinib increases the intracellular concentration of nilotinib, which may explain the observed synergy between these drugs. PMID- 17409348 TI - Survival of genetic subtypes of relapsed myeloma may be modulated by secondary events. PMID- 17409349 TI - The serum-free light chain assay cannot replace 24-hour urine protein estimation in patients with plasma cell dyscrasias. PMID- 17409351 TI - Par1b promotes hepatic-type lumen polarity in Madin Darby canine kidney cells via myosin II- and E-cadherin-dependent signaling. AB - Kidney-derived Madin Darby canine kidney (MDCK) cells form lumina at their apices, and target luminal proteins to an intracellular vacuolar apical compartment (VAC) when prevented from polarizing. Hepatocytes, by contrast, organize their luminal surfaces (the bile canaliculi; BC) between their lateral membranes, and, when nonpolarized, they display an intracellular luminal compartment that is distinct from the VACs of MDCK cells. Overexpression of the serine/threonine kinase Par1b/EMK1/MARK2 induces BC-like lateral lumina and a hepatic-type intracellular luminal compartment in MDCK cells, suggesting a role for Par1b in the branching decision between kidney- and hepatic-type epithelial phenotypes. Here, we report that Par1b promotes lateral lumen polarity in MDCK cells independently of Ca(2+)-mediated cell-cell adhesion by inhibiting myosin II in a rho kinase-dependent manner. Polarization was inhibited by E-cadherin depletion but promoted by an adhesion-defective E-cadherin mutant. By contrast, apical surface formation in control MDCK cells required Ca(2+)-dependent cell cell adhesion, but it occurred in the absence of E-cadherin. We propose that E cadherin, when in an adhesion-incompetent state at the lateral domain, serves as targeting patch for the establishment of lateral luminal surfaces. E-cadherin depletion also reverted the hepatic-type intracellular luminal compartment in Par1b-MDCK cells to VACs characteristic of control MDCK cells, indicating a novel link between E-cadherin and luminal protein targeting. PMID- 17409352 TI - FAK is required for TGFbeta-induced JNK phosphorylation in fibroblasts: implications for acquisition of a matrix-remodeling phenotype. AB - Transforming growth factor beta (TGFbeta) plays a critical role in connective tissue remodeling by fibroblasts during development, tissue repair, and fibrosis. We investigated the molecular pathways in the transmission of TGFbeta signals that lead to features of connective tissue remodeling, namely formation of an alpha-smooth muscle actin (alpha-SMA) cytoskeleton, matrix contraction, and expression of profibrotic genes. TGFbeta causes the activation of focal adhesion kinase (FAK), leading to JNK phosphorylation. TGFbeta induces JNK-dependent actin stress fiber formation, matrix contraction, and expression of profibrotic genes in fak+/+, but not fak-/-, fibroblasts. Overexpression of MEKK1, a kinase acting upstream of JNK, rescues TGFbeta responsiveness of JNK-dependent transcripts and actin stress fiber formation in FAK-deficient fibroblasts. Thus we propose a FAK MEKK1-JNK pathway in the transmission of TGFbeta signals leading to the control of alpha-SMA cytoskeleton reorganization, matrix contraction, and profibrotic gene expression and hence to the physiological and pathological effects of TGFbeta on connective tissue remodeling by fibroblasts. PMID- 17409353 TI - The activity of Pax3 and Zic1 regulates three distinct cell fates at the neural plate border. AB - In Xenopus, the neural plate border gives rise to at least three cell populations: the neural crest, the preplacodal ectoderm, and the hatching gland. To understand the molecular mechanisms that regulate the formation of these lineages, we have analyzed the role of two transcription factors, Pax3 and Zic1, which are among the earliest genes activated in response to neural plate border inducing signals. At the end of gastrulation, Pax3 and Zic1 are coexpressed in the neural crest forming region. In addition, Pax3 is expressed in progenitors of the hatching gland, and Zic1 is detected in the preplacodal ectoderm. Using gain of function and knockdown approaches in whole embryos and animal explants, we demonstrate that Pax3 and Zic1 are necessary and sufficient to promote hatching gland and preplacodal fates, respectively, whereas their combined activity is essential to specify the neural crest. Moreover, we show that by manipulating the levels of Pax3 and Zic1 it is possible to shift fates among these cells. These findings provide novel information on the mechanisms regulating cell fate decisions at the neural plate border. PMID- 17409354 TI - The peroxiredoxin Tpx1 is essential as a H2O2 scavenger during aerobic growth in fission yeast. AB - Peroxiredoxins are known to interact with hydrogen peroxide (H(2)O(2)) and to participate in oxidant scavenging, redox signal transduction, and heat-shock responses. The two-cysteine peroxiredoxin Tpx1 of Schizosaccharomyces pombe has been characterized as the H(2)O(2) sensor that transduces the redox signal to the transcription factor Pap1. Here, we show that Tpx1 is essential for aerobic, but not anaerobic, growth. We demonstrate that Tpx1 has an exquisite sensitivity for its substrate, which explains its participation in maintaining low steady-state levels of H(2)O(2). We also show in vitro and in vivo that inactivation of Tpx1 by oxidation of its catalytic cysteine to a sulfinic acid is always preceded by a sulfinic acid form in a covalently linked dimer, which may be important for understanding the kinetics of Tpx1 inactivation. Furthermore, we provide evidence that a strain expressing Tpx1.C169S, lacking the resolving cysteine, can sustain aerobic growth, and we show that small reductants can modulate the activity of the mutant protein in vitro, probably by supplying a thiol group to substitute for cysteine 169. PMID- 17409355 TI - Active Arf6 recruits ARNO/cytohesin GEFs to the PM by binding their PH domains. AB - ARNO is a soluble guanine nucleotide exchange factor (GEF) for the Arf family of GTPases. Although in biochemical assays ARNO prefers Arf1 over Arf6 as a substrate, its localization in cells at the plasma membrane (PM) suggests an interaction with Arf6. In this study, we found that ARNO activated Arf1 in HeLa and COS-7 cells resulting in the recruitment of Arf1 on to dynamic PM ruffles. By contrast, Arf6 was activated less by ARNO than EFA6, a canonical Arf6 GEF. Remarkably, Arf6 in its GTP-bound form recruited ARNO to the PM and the two proteins could be immunoprecipitated. ARNO binding to Arf6 was not mediated through the catalytic Sec7 domain, but via the pleckstrin homology (PH) domain. Active Arf6 also bound the PH domain of Grp1, another ARNO family member. This interaction was direct and required both inositol phospholipids and GTP. We propose a model of sequential Arf activation at the PM whereby Arf6-GTP recruits ARNO family GEFs for further activation of other Arf isoforms. PMID- 17409356 TI - Mitotic chromosome biorientation in fission yeast is enhanced by dynein and a minus-end-directed, kinesin-like protein. AB - Chromosome biorientation, the attachment of sister kinetochores to sister spindle poles, is vitally important for accurate chromosome segregation. We have studied this process by following the congression of pole-proximal kinetochores and their subsequent anaphase segregation in fission yeast cells that carry deletions in any or all of this organism's minus end-directed, microtubule-dependent motors: two related kinesin 14s (Pkl1p and Klp2p) and dynein. None of these deletions abolished biorientation, but fewer chromosomes segregated normally without Pkl1p, and to a lesser degree without dynein, than in wild-type cells. In the absence of Pkl1p, which normally localizes to the spindle and its poles, the checkpoint that monitors chromosome biorientation was defective, leading to frequent precocious anaphase. Ultrastructural analysis of mutant mitotic spindles suggests that Pkl1p contributes to error-free biorientation by promoting normal spindle pole organization, whereas dynein helps to anchor a focused bundle of spindle microtubules at the pole. PMID- 17409358 TI - Cost-effectiveness acceptability curves in the dock: case not proven? PMID- 17409357 TI - Receptor complexes cotransported via polarized endocytic pathways form clusters with distinct organizations. AB - Previously, FRET confocal microscopy has shown that polymeric IgA-receptor (pIgA R) is distributed in a clustered manner in apical endosomes. To test whether different membrane-bound components form clusters during membrane trafficking, live-cell quantitative FRET was used to characterize the organization of pIgA-R and transferrin receptor (TFR) in endocytic membranes of polarized MDCK cells upon internalization of donor- and acceptor-labeled ligands. We show that pIgA-R and TFR complexes form increasingly organized clusters during cotransport from basolateral to perinuclear endosomes. The organization of these receptor clusters in basolateral versus perinuclear/apical endosomes is significantly different; the former showing a mixed random/clustered distribution while the latter highly organized clusters. Our results indicate that although both perinuclear and apical endosomes comprise pIgA-R and TFR clusters, their E% levels are significantly different suggesting that these receptors are packed into clusters in a distinct manner. The quantitative FRET-based assay presented here suggests that different receptor complexes form clusters, with diverse levels of organization, while being cotransported via the polarized endocytic pathways. PMID- 17409359 TI - The acceptability of acceptability curves: comments on Groot Koerkamp and others. PMID- 17409360 TI - Medical decision making and electronic publishing. PMID- 17409361 TI - Limitations of acceptability curves for presenting uncertainty in cost effectiveness analysis. AB - Clinical journals increasingly illustrate uncertainty about the cost and effect of health care interventions using cost-effectiveness acceptability curves (CEACs). CEACs present the probability that each competing alternative is optimal for a range of values of the cost-effectiveness threshold. The objective of this article is to demonstrate the limitations of CEACs for presenting uncertainty in cost-effectiveness analyses. These limitations arise because the CEAC is unable to distinguish dramatically different joint distributions of incremental cost and effect. A CEAC is not sensitive to any change of the incremental joint distribution in the upper left and lower right quadrants of the cost effectiveness plane; neither is it sensitive to radial shift of the incremental joint distribution in the upper right and lower left quadrants. As a result, CEACs are ambiguous to risk-averse policy makers, inhibit integration with risk attitude, hamper synthesis with other evidence or opinions, and are unhelpful to assess the need for more research. Moreover, CEACs may mislead policy makers and can incorrectly suggest medical importance. Both for guiding immediate decisions and for prioritizing future research, these considerable drawbacks of CEACs should make us rethink their use in communicating uncertainty. As opposed to CEACs, confidence and credible intervals do not conflate magnitude and precision of the net benefit of health care interventions. Therefore, they allow (in)formal synthesis of study results with risk attitude and other evidence or opinions. Presenting the value of information in addition to these intervals allows policy makers to evaluate the need for more empirical research. PMID- 17409362 TI - Value of information on preference heterogeneity and individualized care. AB - BACKGROUND: Cost-effectiveness analysis traditionally focuses on identifying when treatments are cost-effective based on their average benefits and costs in the population. However, there may be considerable value in identifying when treatments are cost-effective for individual patients given their preferences or other personal attributes. OBJECTIVES: To present a theoretical framework to assess the potential value of identifying cost-effective treatments for individual patients given their preferences and to compare the value of individualized treatment decisions with the value of treatment decisions based on traditional population-level cost-effectiveness analysis. Methods . The authors calculate the expected value of individualized care (EVIC), which represents the potential value of providing physicians information on the preferences of individual patients, such as quality-of-life (QOL) weights, so as to make individualized treatment decisions. They also show how EVIC varies with insurance structures that do not internalize relative costs of treatments. They illustrate this theory using an example in which physicians make treatment choices for 65 year-old prostate cancer patients. RESULTS: The value of identifying cost effective treatments at the individual level for 65-year-old prostate cancer patients in the United States is about $70 million annually. This is more than 100 times the $0.7 million annual value of identifying the cost-effective treatment on average for this population. However, failure to internalize costs almost eliminates the value of individualized care. CONCLUSIONS: The value of individualizing care can be far greater than the value of improved decision making at the group level. However, this can vary immensely with insurance. EVIC can provide a guide as to when the high value of individualized care may make population-level decision making especially at risk of providing poor guidance for coverage decisions. Future studies of the value of individualized care should also consider baseline levels of individualization of care. PMID- 17409363 TI - Efficiency, equity, and budgetary policies: informing decisions using mathematical programming. AB - The standard decision rules of cost-effectiveness analysis either require the decision maker to set a threshold willingness to pay for additional health care or to set an overall fixed budget. In practice, neither are generally taken, but instead an arbitrary decision rule is followed that may not be consistent with the overall budget, lead to an allocation of resources that is less than optimal, and is unable to identify the program that should be displaced at the margin. Recent work has shown how mathematical programming can be used as a generalization of the standard decision rules. The authors extend the use of mathematical programming, first to incorporate more complex budgetary rules about when expenditure can be incurred, and show the opportunity loss, in terms of health benefit forgone, of each budgetary policy. Second, the authors demonstrate that indivisibility in a patient population can be regarded as essentially a concern for horizontal equity and represent this and other equity concerns as constraints in the program. Third, the authors estimate the different opportunity costs of a range of equity concerns applied to particular patient populations, and when imposed on all patient populations. They apply this framework of analysis to a realistic and policy-relevant problem. PMID- 17409364 TI - Costs of rheumatoid arthritis: new estimates from the human capital method and comparison to the willingness-to-pay method. AB - BACKGROUND: Individuals' valuation of changes in health states in monetary terms have been measured by examining changes in the direct and indirect costs of disease and by the willingness-to-pay (WTP) methodology. METHODS: In 2002, a 2 part study was conducted in Quebec. In one part of the study, 121 rheumatoid arthritis (RA) patients from the McGill University Health Centre were mailed the Stanford Cost Assessment Questionnaire, which enabled the elicitation of direct costs and indirect costs, according to the friction cost and the human capital methods. The other part was a phone survey conducted in a representative sample of the general population and in the same sample of patients, aiming to elicit the societal WTP for a complete cure of RA in the context of 2 different scenarios: a public coverage or private insurance. These estimates were then compared. RESULTS: Estimates of the cost of illness of RA ranged from 11,717 to 28,498 Canadian Dollars (CAD) depending on the method. These estimates are higher than those previously published in Canada from the 1990s, which is partly due to the recent and costly biological therapies and to a change in the measurement of productivity losses. These estimates are somewhat lower than the societal WTP elicited from the WTP survey, that is, 26,717 and 36,817 CAD per RA case, depending on the public or private health insurance context in which the cure would be available. CONCLUSION: Given that neither method is ideal, data from both methods would provide an important sensitivity analysis when monetary estimates of health state changes are required. PMID- 17409365 TI - Decision to implant a cardioverter defibrillator after myocardial infarction: the role of ejection fraction v. other risk factor markers. AB - BACKGROUND: The Multicenter Automatic Defibrillator Implantation Trial showed that in post-myocardial infarction patients with a left ventricular ejection fraction (EF) 0.30, an implantable cardioverter defibrillator (ICD) resulted in a 31% relative reduction in the risk of death when compared with a conventional therapy group. Whether further refinement in risk estimation could be achieved with additional clinical testing to qualify patients for primary prevention with ICDs remains problematic. METHODS: The authors analyzed Cardiac Arrhythmia Suppression Trial registry data to estimate sensitivity and specificity of EF, ventricular premature frequency, and nonsustained ventricular tachycardia for predicting death. They combined the results with similar data from the literature and used summarizing receiver operating characteristic (meta-ROC) curves to estimate overall operational values for sensitivity and specificity for each clinical test. They estimated aggregate values for prior probability to project risks when tests were used singly and in combination. RESULTS: The authors used arrhythmia markers and heart rate variability to further stratify low-EF patients (prior risk = 20.3%); proportionately, 20.4% were predicted at high risk (>30%) and 40.5% at low risk (<10%). When heart rate variability is normal, those at high risk reduced proportionately to 9.2%, and those at low risk increased to 51.6%. CONCLUSIONS: The combined use of noninvasive markers for arrhythmia substrate and altered autonomic tone can improve risk stratification in low EF without optimal beta-block therapy, whereas for those with optimal beta-block therapy, markers for arrhythmia substrate alone work. Ancillary use of electrophysiologic stimulation can improve results. PMID- 17409366 TI - Cost-effectiveness of echocardiography to identify intracardiac thrombus among patients with first stroke or transient ischemic attack. AB - BACKGROUND AND PURPOSE: Echocardiography to select stroke patients for targeted treatments, such as anticoagulation (AC), to reduce recurrent stroke risk is controversial. The authors' objective was to evaluate the cost-effectiveness of imaging strategies that use transthoracic (TTE) and transesophageal (TEE) echocardiography for identifying intracardiac thrombus in new stroke patients. METHODS: Model-based cost-effectiveness analysis of 7 echocardiographic imaging strategies and 2 nontesting strategies with model parameters based on systematic evidence review related to effectiveness of echocardiography in newly diagnosed ischemic stroke patients (white males aged 65 years in base case). Primary outcome was cost per quality-adjusted life year (QALY). RESULTS: All strategies containing TTE were dominated by others and were eliminated from the analysis. Assuming that AC reduces recurrent stroke risk from intracardiac thrombus by 43% over 1 year, TEE generated a cost per QALY of $137,000 (relative to standard treatment) among patients with 5% thrombus prevalence. Cost per QALY dropped to $50,000 in patients with at least 15% intracardiac thrombus prevalence, or, if an 86% relative risk reduction with AC is assumed, in patients with thrombus prevalence of at least 6%. Probabilistic analyses indicate considerable uncertainty around the cost-effectiveness of echocardiography across a wide range of intracardiac thrombus prevalence (pretest probability). CONCLUSIONS: Current evidence on cost-effectiveness is insufficient to justify widespread use of echocardiography in stroke patients. Additional research on recurrent stroke risk in patients with intracardiac thrombus and on the efficacy of AC in reducing that risk may contribute to a better understanding of the circumstances under which echocardiography will be cost-effective. PMID- 17409367 TI - A real options approach to watchful waiting: theory and an illustration. AB - Watchful waiting is a form of clinical management under which immediate curative treatment is not given. Instead, the patient undergoes a period of observation during which periodic tests monitor the progression of the illness. Hitherto, little attention has been given to how such patient management should be modeled from an economic perspective. Watchful waiting preserves an "option" to start treatment some time in the future, and evaluating a watchful waiting regime therefore has close analogies with pricing a financial option in the derivatives market. This article demonstrates how the methods used to price financial options can be used to decide when to pursue a watchful waiting strategy for a particular patient. The principles of option pricing are illustrated with the example of abdominal aortic aneurysm. A simple trinomial model of disease progression is used, in which patients are periodically monitored, and their health state can remain unchanged, deteriorate, or improve. Backward induction is used to solve the model at each period, with optimal treatment recommendations depending on the current health state. At very low levels of expected net benefits, the patient is discharged. At high levels, the patient is treated immediately. At intermediate levels, watchful waiting continues. The authors argue that option pricing methods offer important insights into the evaluation of a watchful waiting strategy. The methods also have potential applications in other domains of medical care. PMID- 17409368 TI - Patients derogate physicians who use a computer-assisted diagnostic aid. AB - OBJECTIVE: To ascertain whether a physician who uses a computer-assisted diagnostic support system (DSS) would be rated less capable than a physician who does not. METHOD: Students assumed the role of a patient with a possible ankle fracture (experiment 1) or a possible deep vein thrombosis (experiment 2). They read a scenario that described an interaction with a physician who used no DSS, one who used an unspecified DSS, or one who used a DSS developed at a prestigious medical center. Participants were then asked to rate the interaction on 5 criteria, the most important of which was the diagnostic ability of the physician. In experiment 3, 74 patients in the waiting room of a clinic were randomly assigned to the same 3 types of groups as used in experiment 1. In experiment 4, 131 3rd- and 4th-year medical students read a scenario of a physician-patient interaction and were randomly assigned to 1 of 4 groups: the physician used no DSS, heeded the recommendation of a DSS, defied a recommendation of a DSS by treating in a less aggressive manner, or defied a recommendation of a DSS by treating in a more aggressive manner . RESULTS: The participants always deemed the physician who used no decision aid to have the highest diagnostic ability. CONCLUSION: Patients may surmise that a physician who uses a DSS is not as capable as a physician who makes the diagnosis with no assistance from a DSS. PMID- 17409369 TI - The influence of irrelevant anchors on the judgments and choices of doctors and patients. AB - BACKGROUND: Little research has examined how anchor numbers affect choice, despite several decades of research showing that judgments typically and robustly assimilate toward irrelevant anchors. METHODS: In one experiment, HIV-positive patients (N = 99) judged the chances that sexual partners would become infected with HIV after sex using a defective condom and then indicated their choices of remedial action. In a second experiment, Iowa physicians (N =191) rated the chances that hypothetical patients had a pulmonary embolism and then formulated a treatment plan. RESULTS: Irrelevant anchor numbers dramatically affected judgments by HIV-infected patients of the chances of HIV infection after a condom broke during sex (43% v. 64% in the low- and high-anchor conditions, respectively) and judgments by doctors of the chances of pulmonary embolism (23% v. 53%, respectively). Despite large anchoring effects in judgement, treatment choices did not differ between low-and high-anchor conditions. Accountability did not reduce the anchoring bias in the doctors' judgments. DISCUSSION: The practical implications of anchoring for risk judgments are potentially large, but the bias may be less relevant to treatment choices. The findings suggest that the theoretical underpinnings of the anchoring bias may be more complex than previously thought. PMID- 17409370 TI - Correction: interpretation of random effects meta-analysis in decision models. PMID- 17409371 TI - The Stroke Outcomes and Neuroimaging of Intracranial Atherosclerosis (SONIA) trial. AB - BACKGROUND: Transcranial Doppler ultrasound (TCD) and magnetic resonance angiography (MRA) can identify intracranial atherosclerosis but have not been rigorously validated against the gold standard, catheter angiography. The WASID trial (Warfarin Aspirin Symptomatic Intracranial Disease) required performance of angiography to verify the presence of intracranial stenosis, allowing for prospective evaluation of TCD and MRA. The aims of Stroke Outcomes and Neuroimaging of Intracranial Atherosclerosis (SONIA) trial were to define abnormalities on TCD/MRA to see how well they identify 50 to 99% intracranial stenosis of large proximal arteries on catheter angiography. STUDY DESIGN: SONIA standardized the performance and interpretation of TCD, MRA, and angiography. Study-wide cutpoints defining positive TCD/MRA were used. Hard copy TCD/MRA were centrally read, blind to the results of angiography. RESULTS: SONIA enrolled 407 patients at 46 sites in the United States. For prospectively tested noninvasive test cutpoints, positive predictive values (PPVs) and negative predictive values (NPVs) were TCD, PPV 36% (95% CI: 27 to 46); NPV, 86% (95% CI: 81 to 89); MRA, PPV 59% (95% CI: 54 to 65); NPV, 91% (95% CI: 89 to 93). For cutpoints modified to maximize PPV, they were TCD, PPV 50% (95% CI: 36 to 64), NPV 85% (95% CI: 81 to 88); MRA PPV 66% (95% CI: 58 to 73), NPV 87% (95% CI: 85 to 89). For each test, a characteristic performance curve showing how the predictive values vary with a changing test cutpoint was obtained. CONCLUSIONS: Both transcranial Doppler ultrasound and magnetic resonance angiography noninvasively identify 50 to 99% intracranial large vessel stenoses with substantial negative predictive value. The Stroke Outcomes and Neuroimaging of Intracranial Atherosclerosis trial methods allow transcranial Doppler ultrasound and magnetic resonance angiography to reliably exclude the presence of intracranial stenosis. Abnormal findings on transcranial Doppler ultrasound or magnetic resonance angiography require a confirmatory test such as angiography to reliably identify stenosis. PMID- 17409372 TI - Modulation of human arterial tone during pregnancy: the effect of the bioactive metabolite sphingosine-1-phosphate. AB - Sphingosine-1-phosphate (S1P) is a potent bioactive lipid that has been implicated in cardiovascular disease. The objective of the present study was to determine the vasoactive effects and underlying mechanisms of S1P on adult human maternal arteries. The isometric tensions of the omental and myometrial arteries isolated from normal pregnant women at term were assessed in response to incremental doses of S1P in the presence or absence of the nitric oxide (NO) synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). The putative involvement of Rho-associated kinases (ROCKs) in intact arteries and in those permeabilized with alpha-toxin, to study agonist-dependent calcium-sensitization, was assessed with the inhibitor Y27632. Real-time RT-PCR established the presence of mRNA encoding the S1P receptors (S1P(1) to (3)), previously known as endothelial differentiation gene receptors (EDG1, 3 and 5), in both artery types. S1P induced a dose-dependent increase in the isometric tension of all the arteries. Y27632 reduced constriction due to S1P in intact arteries and reduced S1P-induced sensitization of contraction to submaximal activating Ca(2+) in permeabilized arteries. L-NAME also modulated S1P vasoactive responses in a tissue-specific manner. Two subgroups of omental arteries were identified, one of which utilizes the NO pathway. In myometrial arteries, S1P evoked oscillatory constrictions, whereas pretreatment with L-NAME resulted in only tonic constrictions of unaltered peak magnitude. The prominent vasoactive actions of S1P in the maternal arteries of pregnant women are modulated by inhibitors of ROCKs and NO bioavailability. The subtle tissue-specific functional differences in the modulation of S1P actions by NO have important implications for vascular tone regulation by this bioactive circulatory metabolite during pregnancy. PMID- 17409373 TI - Constitutive steroidogenesis in ovine large luteal cells may be mediated by tonically active protein kinase A. AB - The mechanisms responsible for the increased basal rates of progesterone secretion from large steroidogenic luteal cells (LLC) relative to small steroidogenic luteal cells (SLC) have not been clearly defined. To determine if protein kinase A (PKA) is tonically active in LLC, the adenylate cyclase activator forskolin and a specific PKA inhibitor (PKI) were utilized in a 2 x 2 factorial treatment with each steroidogenic cell type. Progesterone and cAMP production were quantified after the different treatments. In addition, the effects of the treatments on the concentrations and relative phosphorylation status of the steroidogenic acute regulatory (STAR) protein in the two cell types were determined as a measure of PKA activity. Treatment with PKI blocked forskolin-induced increases in progesterone secretion by SLC without affecting the production of cAMP. The treatment of LLC with PKI significantly decreased basal progesterone secretion in the presence or absence of forskolin, indicating that the high level of steroidogenesis in this cell type requires PKA activity. There were no differences in the steady-state concentrations of STAR protein in either cell type after treatment. However, the percentage of relative STAR phosphorylation was higher in the LLC than in SLC, and PKI treatment significantly decreased the phosphorylation of STAR in the LLC. The relative phosphorylation status of STAR and the concentrations of progesterone in the media were significantly correlated with the treatments in both cell types. The amount of progesterone secreted per picogram of cAMP was higher in the LLC than in the SLC, and this was accompanied by a significant increase in the ratio of relative STAR phosphorylation to the steady-state concentration of STAR protein. These data are compatible with the theory that LLC are constitutively steroidogenic, partly because they have tonically active PKA. In addition, the phosphorylation of STAR appears to be a primary activity of PKA in both types of ovine steroidogenic luteal cells. PMID- 17409374 TI - Actions of epidermal growth factor receptor/mitogen-activated protein kinase and protein kinase C signaling in granulosa cells from Gallus gallus are dependent upon stage of differentiation. AB - Studies in both mammalian and nonmammalian ovarian model systems have demonstrated that activation of the mitogen-activated protein kinase (MAPK) and protein kinase C (PKC) signaling pathways modulates steroid biosynthesis during follicle development, yet the collective evidence for facilitory versus inhibitory roles of these pathways is inconsistent. The present studies in the hen ovary describe the changing role of MAPK and PKC signaling in the regulation of steroidogenic acute regulatory protein (STAR) expression and progesterone production in undifferentiated granulosa cells collected from prehierarchal follicles prior to follicle selection versus differentiated granulosa from preovulatory follicles subsequent to selection. Treatment of undifferentiated granulosa cells with a selective epidermal growth factor receptor (EGFR) and ERBB4 receptor tyrosine kinase inhibitor (AG1478) both augments FSH receptor (Fshr) mRNA expression and initiates progesterone production. Conversely, selective inhibitors of both EGFR/ERBB4 and MAPK activity attenuate steroidogenesis in differentiated granulosa cells subsequent to follicle selection. In addition, inhibition of PKC signaling with GF109203X augments FSH induced Fshr mRNA plus STAR protein expression and initiates progesterone synthesis in undifferentiated granulosa cells, but inhibits both gonadotropin induced STAR expression and progesterone production in differentiated granulosa. Granulosa cells from the most recently selected (9- to 12-mm) follicle represent a stage of transition as inhibition of MAPK signaling promotes, while inhibition of PKC signaling blocks gonadotropin-induced progesterone production. Collectively, these data describe stage-of-development-related changes in cell signaling whereby the differentiation-inhibiting actions of MAPK and PKC signaling in prehierarchal follicle granulosa cells undergo a transition at the time of follicle selection to become obligatory for gonadotropin-stimulated progesterone production in differentiated granulosa from preovulatory follicles. PMID- 17409375 TI - Impaired progesterone production in Nr5a2+/- mice leads to a reduction in female reproductive function. AB - NR5A2 is an orphan nuclear receptor involved in cholesterol metabolism and embryogenesis. The high level of expression of NR5A2 in the ovary and its involvement in the regulation of steroidogenic gene expression also suggest a role for this transcription factor in female reproductive function. In vivo evidence for a role for NR5A2 in fertility, however, is still lacking. In order to address this possibility, we used Nr5a2+/- mice to demonstrate that heterozygosity for a null mutation of Nr5a2 leads to a decreased fertility in females. Our results indicate that although Nr5a2+/- mice display normal follicular development, ovulation, and estrogen production, they exhibit altered luteal function. More specifically, we show that the reduced reproductive ability of Nr5a2+/- females arises from a reduction in circulating progesterone concentrations and can be rescued by exogenous progesterone supplementation. This study therefore provides the first in vivo evidence for a role of NR5A2 in reproductive function and steroidogenesis. PMID- 17409376 TI - Evaluation of chromosomal risk following intracytoplasmic sperm injection in the mouse. AB - To investigate whether cytogenetic risks occur using the mouse intracytoplasmic sperm injection (ICSI) technique, the incidence of chromosome aberrations was compared in one-cell embryos produced by ICSI technique and those by conventional in vitro fertilization (IVF) technique. Spermatozoa were incubated in TYH medium for 1.5-2 h before IVF insemination. For the ICSI technique, spermatozoa were incubated in five different media: TYH, Hepes-buffered TYH (H-TYH), modified CZB (mCZB), Hepes-buffered mCZB (H-mCZB), and PB1 for 0.5 h, 2-2.5 h, and 6 h before injection into metaphase II oocytes. The incidence of IVF embryos with structural chromosome aberrations was 2%, whereas the occurrence of structural chromosome aberrations in ICSI embryos was dependent on the kind of medium and sperm incubation time. When spermatozoa were incubated in TYH medium for 2 h or more, the aberration rates in the resultant ICSI embryos (4%) were not significantly different from that of IVF embryos. However, there was a significant increase in aberration rates in ICSI embryos derived from spermatozoa that were incubated in other culture conditions (6%-28%). In addition, a time-dependent increase in aberration rates was found in ICSI embryos when H-TYH, H-mCZB, and PB1 were used for sperm incubation. There was no significant difference in incidence of aneuploidy between IVF and ICSI embryos. The chromosome analysis results of one cell embryos were reflected by the performance of postimplantation embryo development. The causal mechanism of chromosome damage in ICSI embryos was discussed in relation to the plasma membrane cholesterol, the acrosome, and in vitro aging of spermatozoa. PMID- 17409377 TI - Vitellogenin and its messenger RNA during ovarian development in the female blue crab, Callinectes sapidus: gene expression, synthesis, transport, and cleavage. AB - Blue crab vitellogenin (VTG) cDNA encodes a precursor that, together with two other Brachyuran VTGs, forms a distinctive cluster within a phylogenetic tree of crustacean VTGs. Using quantitative RT-PCR, we found that VTG was primarily expressed in the hepatopancreas of a vitellogenic female, with minor expression in the ovary. VTG expression in the hepatopancreas correlated with ovarian growth, with a remarkable 8000-fold increase in expression from stage 3 to 4 of ovarian development. In contrast, the VTG levels in the hepatopancreas and hemolymph decreased in stage 4. Western blot analysis and N-terminal sequencing revealed that vitellin is composed of three subunits of approximately 78.5 kDa, 119.42 kDa, and 87.9 kDa. The processing pathway for VTG includes an initial hepatopancreatic cleavage of the primary precursor into approximately 78.5-kDa and 207.3-kDa subunits, both of which are found in the hemolymph. A second cleavage in the ovary splits the approximately 207.3-kDa subunit into approximately 119.4-kDa and approximately 87.9-kDa subunits. The hemolymph VTG profiles of mated and unmated females during ovarian development indicate that early vitellogenesis and ovarian development do not require mating, which may be essential for later stages, as VTG decreased to the basal level at stage 4 in the unmated group but remained high in the mated females. Our results encompass comprehensive overall temporal and spatial aspects of vitellogenesis, which may reflect the reproductive physiology of the female blue crab, e.g., single mating and anecdysis in adulthood. PMID- 17409378 TI - Effects of detergents on the redistribution of gangliosides and GPI-anchored proteins in brain tissue sections. AB - Gangliosides and glycosylphosphatidylinositol (GPI)-anchored proteins contain lipid tails that tether them to the outer side of the cell membrane. This mode of association with the cell membrane enables them to take part in the organization of lipid rafts, but it also permits gangliosides and GPI-anchored proteins to be actively released from one cell and inserted into the membrane of another cell. Recently, we reported that under conditions of lipid raft isolation, Triton X-100 causes significant redistribution of both gangliosides and GPI-anchored proteins. Aiming to find a less disruptive detergent, we evaluated the effects of CHAPS, Saponin, deoxycholic acid, Trappsol, Tween 20, Triton X-100, Brij 96V, Brij 98, and SDS on brain tissue sections. At room temperature, all detergents (1% concentration) extracted significant amounts of both gangliosides and Thy-1. At 4C, the extraction was weaker, but Triton X-100, CHAPS, and deoxycholic acid caused significant redistribution of GD1a and Thy-1 from gray matter into the white matter. Both redistribution and extraction were significantly augmented when sections were incubated with detergents in the presence of primary antibodies. Of the nine tested detergents, none is the ideal choice. However, Brij 96V appears to be able to sufficiently reveal myelin epitopes while causing the least amount of artifacts. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 17409379 TI - Proteome analysis of microdissected formalin-fixed and paraffin-embedded tissue specimens. AB - Targeted proteomics research, based on the enrichment of disease-relevant proteins from isolated cell populations selected from high-quality tissue specimens, offers great potential for the identification of diagnostic, prognostic, and predictive biological markers for use in the clinical setting and during preclinical testing and clinical trials, as well as for the discovery and validation of new protein drug targets. Formalin-fixed and paraffin-embedded (FFPE) tissue collections, with attached clinical and outcome information, are invaluable resources for conducting retrospective protein biomarker investigations and performing translational studies of cancer and other diseases. Combined capillary isoelectric focusing/nano-reversed-phase liquid chromatography separations equipped with nano-electrospray ionization-tandem mass spectrometry are employed for the studies of proteins extracted from microdissected FFPE glioblastoma tissues using a heat-induced antigen retrieval (AR) technique. A total of 14,478 distinct peptides are identified, leading to the identification of 2733 non-redundant SwissProt protein entries. Eighty-three percent of identified FFPE tissue proteins overlap with those obtained from the pellet fraction of fresh-frozen tissue of the same patient. This large degree of protein overlapping is attributed to the application of detergent-based protein extraction in both the cell pellet preparation protocol and the AR technique. PMID- 17409380 TI - Upregulation of vascular endothelial growth factor receptors Flt-1 and Flk-1 following acute spinal cord contusion in rats. AB - To investigate the possible role of vascular endothelial growth factor (VEGF) in the injured spinal cord, we analyzed the distribution and time course of the two tyrosine kinase receptors for VEGF, Flt-1 and Flk-1, in the rat spinal cord following contusion injury using a weight-drop impactor. The semi-quantitative RT PCR analysis of Flt-1 and Flk-1 in the spinal cord showed slight upregulation of these receptors following spinal cord injury. Although mRNAs for Flt-1 and Flk-1 were constitutively expressed in neurons, vascular endothelial cells, and some astrocytes in laminectomy control rats, their upregulation was induced in association with microglia/macrophages and reactive astrocytes in the vicinity of the lesion within 1 day in rats with a contusion injury and persisted for at least 14 days. The spatiotemporal expression of Flt-1 in the contused spinal cord mirrored that of Flk-1 expression. In the early phase of spinal cord injury, upregulation of Flt-1 and Flk-1 mRNA occurred in microglia/macrophages that infiltrated the lesion. In addition, the expression of both receptors increased progressively in reactive astrocytes within the vicinity of the lesion, predominately in the white matter, and almost all reactive astrocytes coexpressed Flt-1 or Flk-1 and nestin. These results suggest that VEGF may be involved in the inflammatory response and the astroglial reaction to contusion injuries of the spinal cord via specific VEGF receptors. PMID- 17409381 TI - Expression and subcellular localization of a 35-kDa carbonic anhydrase IV in a human pancreatic ductal cell line (Capan-1). AB - The high intraluminal concentrations of HCO(3)(-) in the human pancreatic ducts have suggested the existence of a membrane protein supplying the Cl(-)/HCO(3)(-) exchanger. Membrane-bound carbonic anhydrase IV (CA IV) is one of the potential candidates for this protein. The difficulties in isolating human pancreatic ducts have led the authors to study the molecular mechanisms of HCO(3)(-) secretion in cancerous cell lines. In this work, we have characterized the CA IV expressed in Capan-1 cells. A 35-kDa CA IV was detected in cell homogenates and purified plasma membranes. Treatment of purified plasma membranes with phosphatidylinositol-phospholipase-C indicated that this CA IV was not anchored by a glycosylphosphatidylinositol (GPI). In contrast, its detection on purified plasma membranes by an antibody specifically directed against the carboxyl terminus of human immature GPI-anchored CA IV indicated that it was anchored by a C-terminal hydrophobic segment. Immunoelectron microscopy and double-labeling immunofluorescence revealed that this CA IV was present on apical plasma membranes, and in the rough endoplasmic reticulum, the endoplasmic reticulum Golgi intermediate compartment, the Golgi complex, and secretory granules, suggesting its transport via the classical biosynthesis/secretory pathway. The expression in Capan-1 cells of a 35-kDa CA IV anchored in the apical plasma membrane through a hydrophobic segment, as is the case in the healthy human pancreas, should make the study of its role in pancreatic HCO(3)(-) secretion easier. PMID- 17409382 TI - In-depth analysis of the adipocyte proteome by mass spectrometry and bioinformatics. AB - Adipocytes are central players in energy metabolism and the obesity epidemic, yet their protein composition remains largely unexplored. We investigated the adipocyte proteome by combining high accuracy, high sensitivity protein identification technology with subcellular fractionation of nuclei, mitochondria, membrane, and cytosol of 3T3-L1 adipocytes. We identified 3,287 proteins while essentially eliminating false positives, making this one of the largest high confidence proteomes reported to date. Comprehensive bioinformatics analysis revealed that the adipocyte proteome, despite its specialized role, is very complex. Comparison with microarray data showed that the mRNA abundance of detected versus non-detected proteins differed by less than 2-fold and that proteomics covered as large a proportion of the insulin signaling pathway. We used the Endeavour gene prioritization algorithm to associate a number of factors with vesicle transport in response to insulin stimulation, a key function of adipocytes. Our data and analysis can serve as a model for cellular proteomics. The adipocyte proteome is available as supplemental material and from the Max Planck Unified Proteome database. PMID- 17409383 TI - Exploring molecular oxygen pathways in Hansenula polymorpha copper-containing amine oxidase. AB - The accessibility of large substrates to buried enzymatic active sites is dependent upon the utilization of proteinaceous channels. The necessity of these channels in the case of small substrates is questionable because diffusion through the protein matrix is often assumed. Copper amine oxidases contain a buried protein-derived quinone cofactor and a mononuclear copper center that catalyze the conversion of two substrates, primary amines and molecular oxygen, to aldehydes and hydrogen peroxide, respectively. The nature of molecular oxygen migration to the active site in the enzyme from Hansenula polymorpha is explored using a combination of kinetic, x-ray crystallographic, and computational approaches. A crystal structure of H. polymorpha amine oxidase in complex with xenon gas, which serves as an experimental probe for molecular oxygen binding sites, reveals buried regions of the enzyme suitable for transient molecular oxygen occupation. Calculated O(2) free energy maps using copper amine oxidase crystal structures in the absence of xenon correspond well with later experimentally observed xenon sites in these systems, and allow the visualization of O(2) migration routes of differing probabilities within the protein matrix. Site-directed mutagenesis designed to block individual routes has little effect on overall k(cat)/K(m) (O(2)), supporting multiple dynamic pathways for molecular oxygen to reach the active site. PMID- 17409384 TI - The lysine-specific demethylase 1 is required for cell proliferation in both p53 dependent and -independent manners. AB - The lysine-specific demethylase 1 (LSD1), a component of several histone deacetylase complexes, plays an important role in chromatin remodeling and transcriptional regulation. Here, we generated multiple cell lines in which LSD1 is inducibly expressed or knocked down and found that LSD1 is required for cell proliferation. In addition, we found that deficiency in LSD1 leads to a partial cell cycle arrest in G(2)/M and sensitizes cells to growth suppression induced by DNA damage or MDM2 inhibition in a p53-dependent manner. We also showed that LSD1 deficiency delays p53 stabilization induced by DNA damage, leading to a delayed induction of p21 and MDM2. Finally, we performed a microarray study and identified several novel LSD1 target genes, including S100A8, which encodes a calcium-binding protein, and DEK, a proto-oncogene. Taken together, we uncovered that LSD1 has a pro-oncogenic function by modulating pro-survival gene expression and p53 transcriptional activity. PMID- 17409385 TI - Identification, molecular cloning, and characterization of the sixth subunit of human transcription factor TFIIIC. AB - TFIIIC in yeast and humans is required for transcription of tRNA and 5 S RNA genes by RNA polymerase III. In the yeast Saccharomyces cerevisiae, TFIIIC is composed of six subunits, five of which are conserved in humans. We report the identification, molecular cloning, and characterization of the sixth subunit of human TFIIIC, TFIIIC35, which is related to the smallest subunit of yeast TFIIIC. Human TFIIIC35 does not contain the phosphoglycerate mutase domain of its yeast counterpart, and these two proteins display only limited homology within a 34 amino acid domain. Homologs of the sixth TFIIIC subunit are also identified in other eukaryotes, and their phylogenic evolution is analyzed. Affinity-purified human TFIIIC from an epitope-tagged TFIIIC35 cell line is active in binding to and in transcription of the VA1 gene in vitro. Furthermore, TFIIIC35 specifically interacts with the human TFIIIC subunits TFIIIC63 and, to a lesser extent, TFIIIC90 in vitro. Finally, we determined a limited region in the smallest subunit of yeast TFIIIC that is sufficient for interacting with the yeast TFIIIC subunit ScTfc1 (orthologous to TFIIIC63) and found it to be adjacent to and overlap the 34-amino acid domain that is conserved from yeast to humans. PMID- 17409386 TI - The Rab5 activator ALS2/alsin acts as a novel Rac1 effector through Rac1 activated endocytosis. AB - Mutations in the ALS2 gene cause a number of recessive motor neuron diseases, indicating that the ALS2 protein (ALS2/alsin) is vital for motor neurons. ALS2 acts as a guanine nucleotide exchange factor (GEF) for Rab5 (Rab5GEF) and is involved in endosome dynamics. However, the spatiotemporal regulation of the ALS2 mediated Rab5 activation is unclear. Here we identified an upstream activator for ALS2 and showed a functional significance of the ALS2 activation in endosome dynamics. ALS2 preferentially interacts with activated Rac1. In the cells activated Rac1 recruits cytoplasmic ALS2 to membrane ruffles and subsequently to nascent macropinosomes via Rac1-activated macropinocytosis. At later endocytic stages macropinosomal ALS2 augments fusion of the ALS2-localized macropinosomes with the transferrin-positive endosomes, depending on the ALS2-associated Rab5GEF activity. These results indicate that Rac1 promotes the ALS2 membranous localization, thereby rendering ALS2 active via Rac1-activated endocytosis. Thus, ALS2 is a novel Rac1 effector and is involved in Rac1-activated macropinocytosis. All together, loss of ALS2 may perturb macropinocytosis and/or the following membrane trafficking, which gives rise to neuronal dysfunction in the ALS2-linked motor neuron diseases. PMID- 17409387 TI - Histone deacetylase inhibitor trichostatin A sustains sodium pervanadate-induced NF-kappaB activation by delaying IkappaBalpha mRNA resynthesis: comparison with tumor necrosis factor alpha. AB - NF-kappaB is a crucial transcription factor tightly regulated by protein interactions and post-translational modifications, like phosphorylation and acetylation. A previous study has shown that trichostatin A (TSA), a histone deacetylase inhibitor, potentiates tumor necrosis factor (TNF) alpha-elicited NF kappaB activation and delays IkappaBalpha cytoplasmic reappearance. Here, we demonstrated that TSA also prolongs NF-kappaB activation when induced by the insulino-mimetic pervanadate (PV), a tyrosine phosphatase inhibitor that initiates an atypical NF-kappaB signaling. This extension is similarly correlated with delayed IkappaBalpha cytoplasmic reappearance. However, whereas TSA causes a prolonged IKK activity when added to TNFalpha, it does not when added to PV. Instead, quantitative reverse transcriptase-PCR revealed a decrease of ikappabalpha mRNA level after TSA addition to PV stimulation. This synthesis deficit of the inhibitor could explain the sustained NF-kappaB residence in the nucleus. In vivo analysis by chromatin immunoprecipitation assays uncovered that, for PV induction but not for TNFalpha, the presence of TSA provokes several impairments on the ikappabalpha promoter: (i) diminution of RNA Pol II recruitment; (ii) reduced acetylation and phosphorylation of histone H3-Lys(14) and -Ser(10), respectively; (iii) decreased presence of phosphorylated p65 Ser(536); and (iv) reduction of IKKalpha binding. The recruitment of these proteins on the icam-1 promoter, another NF-kappaB-regulated gene, is not equally affected, suggesting a promoter specificity of PV with TSA stimulation. Taken together, these data suggest that TSA acts differently depending on the NF-kappaB pathway and the targeted promoter in question. This indicates that one overall histone deacetylase role is to inhibit NF-kappaB activation by molecular mechanisms specific of the stimulus and the promoter. PMID- 17409388 TI - Origin of fatigue in multiple sclerosis: review of the literature. AB - Fatigue is one of the most common and most disabling symptoms of multiple sclerosis (MS). Although numerous studies have tried to reveal it, no definite pathogenesis factor behind this fatigue has been identified. Fatigue may be directly related to the disease mechanisms (primary fatigue) or may be secondary to non-disease-specific factors. Primary fatigue may be the result of inflammation, demyelination, or axonal loss. A suggested functional cortical reorganization may result in a higher energy demand in certain brain areas, culminating in an increase of fatigue perception. Higher levels of some immune markers were found in patients with MS-related fatigue, whereas other studies rejected this hypothesis. There may be a disturbance in the neuroendocrine system related to fatigue, but it is not clear whether this is either the result of the interaction with immune activation or the trigger of this process. Fatigue may be secondary to sleep problems, which are frequently present in MS and in their turn result from urinary problems, spasms, pain, or anxiety. Pharmacologic treatment of MS (symptoms) may also provoke fatigue. The evidence for reduced activity as a cause of secondary fatigue in MS is inconsistent. Psychological functioning may at least play a role in the persistence of fatigue. Research did not reach consensus about the association of fatigue with clinical or demographic variables, such as age, gender, disability, type of MS, education level, and disease duration. In conclusion, it is more likely to explain fatigue from a multifactor perspective than to ascribe it to one mechanism. The current evidence on the pathogenesis of primary and secondary fatigue in MS is limited by inconsistency in defining specific aspects of the concept fatigue, by the lack of appropriate assessment tools, and by the use of heterogeneous samples. Future research should overcome these limitations and also include longitudinal designs. PMID- 17409389 TI - Transcranial magnetic stimulation and muscle contraction to enhance stroke recovery: a randomized proof-of-principle and feasibility investigation. AB - OBJECTIVE: To explore the efficacy of repetitive transcranial magnetic stimulation (rTMS) and voluntary muscle contraction (VMC) to improve corticospinal transmission, muscle function, and purposeful movement early after stroke. METHODS: Factorial 2 x 2 randomized single-blind trial. SUBJECTS: n = 27, mean age 75 years, mean 27 days after middle cerebral artery infarct (24 subjects completed outcome measures). PROCEDURE: after baseline measurement (day 1), subjects were randomized to 1 of 4 groups. Treatment was given for the next 8 working days, and outcome was measured on day 10. INTERVENTIONS: (a) Real-rTMS + RealVMC, (b) Real-rTMS + PlaceboVMC, (c) Placebo-rTMS + RealVMC, and (d) Placebo rTMS + PlaceboVMC. Real-rTMS consisted of 200 1-Hz stimuli at 120% motor threshold in 5 blocks of 40 separated by 3 minutes delivered to the lesioned hemisphere. Placebo-rTMS used a dummy coil. In RealVMC, the paretic elbow was repeatedly flexed/extended for 5 minutes. In PlaceboVMC, subjects viewed pairs of drawings of upper limbs and reported their likeness. OUTCOMES: frequency of motor evoked potentials in biceps and triceps, muscle function (torque about elbow), and purposeful movement (Action Research Arm Test). ANALYSIS: group mean changes (outcome - baseline) were compared. RESULTS: In the Real-rTMS + RealVMC group, motor-evoked potential frequency increased 14% for biceps and 20% for triceps, whereas in the Placebo-rTMS + PlaceboVMC group, it decreased 12% for biceps and 6% for triceps. For other groups, there were changes of intermediate values. No meaningful differences were found for secondary outcomes. CONCLUSIONS: A positive trend for motor-evoked potential frequency was found for Real-rTMS + RealVMC, whereas a negative trend for motor-evoked potential frequency was found for Placebo-rTMS + PlaceboVMC. PMID- 17409390 TI - Physiological Cost Index as a proxy measure for the oxygen cost of gait in stroke patients. AB - BACKGROUND: Stroke survivors can exhibit abnormally elevated oxygen consumption during walking. Therapeutic interventions can improve gait deficits and oxygen consumption. A practical measure of oxygen cost is not available. This study tested the usefulness of an indirect index of oxygen cost, the Physiological Cost Index, and the ability of this index to discriminate between healthy adults and stroke survivors. METHODS: The authors studied 17 subjects with stroke and 10 healthy control participants. Participants walked 10 minutes at their chosen comfortable speed on a treadmill. Oxygen consumption and heart rate data were collected. Primary measures were oxygen cost and the Physiological Cost Index. Secondary measures were age and gait speed. RESULTS: The Physiological Cost Index and oxygen cost had a good to excellent correlation (r = .83, P < .001) for subjects with stroke. Both oxygen cost and the Physiological Cost Index were comparable in detecting a significantly abnormal elevation for stroke survivors versus healthy adults (P = .003 and .002, respectively). Age was not correlated with oxygen cost, the Physiological Cost Index, or chosen gait speed. A moderate correlation of gait speed to both the Physiological Cost Index and oxygen cost was found. CONCLUSIONS: The Physiological Cost Index can be used as a proxy index for the oxygen cost of walking in subjects after stroke because it is correlated with oxygen cost and is comparable to oxygen cost in its capability to discriminate between healthy controls and subjects with stroke. The Physiological Cost Index can be performed inexpensively on a routine basis in a clinical environment. PMID- 17409391 TI - Genetic variation at 8q24 as a susceptibility factor for prostate cancer: definitive results from epidemiologic studies? PMID- 17409392 TI - Targeting multiple arms of the apoptotic regulatory machinery. AB - ABT-737 targets Bcl-2/Bcl-xL but not Mcl-1, which confers resistance to this novel agent. Here, we summarize recent findings indicating that Mcl-1 represents a critical determinant of ABT-737 sensitivity and resistance, and that Mcl-1 down regulation by various pharmacologic agents or genetic approaches dramatically increases ABT-737 lethality in diverse malignant cell types. These findings also show that the multidomain proapoptotic proteins Bax and Bak play important functional roles in ABT-737-mediated apoptosis, and that Bak activation is essential in potentiation of ABT-737 lethality by agents that down-regulate Mcl 1. Collectively, these findings suggest a novel therapeutic strategy targeting multiple arms of the apoptotic machinery. PMID- 17409393 TI - Tumor-released microvesicles as vehicles of immunosuppression. AB - Tumor-released microvesicles, or exosomes, which are abundant in the body fluids of patients with cancer, are likely to be involved in tumor progression. We recently showed that microvesicles released by human melanoma and colorectal carcinoma cells can promote the differentiation of monocytes to myeloid-derived suppressor cells which support tumoral growth and immune escape. These findings underscore an important role for these extracellular organelles in remodeling tumor-stromal interactions to promote malignancy. PMID- 17409394 TI - Dual role of inactivating Lef1 mutations in epidermis: tumor promotion and specification of tumor type. AB - The NH(2) terminus of LEF1 is frequently mutated in human sebaceous tumors. To investigate how this contributes to cancer, we did two-stage chemical carcinogenesis on K14DeltaNLef1 transgenic mice, which express NH(2)-terminally truncated Lef1 in the epidermal basal layer. Transgenic mice developed more tumors, more rapidly than littermate controls, even without exposure to tumor promoter. They developed sebaceous tumors, whereas controls developed squamous cell carcinomas. K14DeltaNLef1 epidermis failed to up-regulate p53 and p21 proteins during tumorigenesis or in response to UV irradiation, and this correlated with impaired p14ARF induction. We propose that LEF1 NH(2)-terminal mutations play a dual role in skin cancer, specifying tumor type by inhibiting Wnt signaling and acting as a tumor promoter by preventing induction of p53. PMID- 17409395 TI - PRL-3 down-regulates PTEN expression and signals through PI3K to promote epithelial-mesenchymal transition. AB - PRL-3 is a metastasis-associated phosphatase. We and others have shown that its overexpression increases cell motility and invasiveness. These phenotypic changes are reminiscent of the epithelial-mesenchymal transition (EMT) that occurs during embryonic development and oncogenesis. The EMT is a complex process that converts epithelia into migratory mesenchymal cells. We here attempt to unravel the underlying mechanistic basis of these phenomena. HeLa cells transiently expressing EGFP-PRL-3 (HeLa-PRL-3) exhibit reduced levels of paxillin. Similarly, Chinese hamster ovary cells stably expressing myc-PRL-3 (CHO-PRL-3) also show marked reductions in paxillin, phosphorylated paxillin-Tyr(31), and vinculin at focal adhesion complexes and notable reductions in the levels of RhoA-GTP, Rac1 GTP, and filamentous-actin filaments. DLD-1 human colorectal cancer cells engineered to express EGFP-PRL-3 (DLD-1-PRL-3) underwent changes consistent with EMT. In these cells, PRL-3 activates Akt and inactivates glycogen synthase kinase 3beta as assessed by phosphospecific antibodies. PRL-3 up-regulates mesenchymal markers fibronectin and Snail and down-regulates epithelial markers E-cadherin, gamma-catenin (plakoglobin), and integrin beta(3), which are major effectors in the EMT pathway. The changes in these EMT characteristics brought about by PRL-3 can be abrogated by the phosphoinositide 3-kinase (PI3K) inhibitor LY294002, implying that PRL-3 acts upstream of PI3K and could play an initiating role to trigger the EMT switch during cancer metastasis. In addition, PRL-3 can down regulate phosphatase and tensin homologue deleted on chromosome 10, which is an important antagonist of PI3K, further reinforcing PI3K/Akt function in PRL-3 triggered EMT. Catalytically inactive PRL-3 (C104S) was impaired in the above PRL 3-mediated events, indicating that these properties require phosphatase activity. Targeting PRL-3 may thus be a useful strategy to impede cancer cell invasion and metastasis. PMID- 17409396 TI - Mammalian tumor xenografts induce neovascularization in zebrafish embryos. AB - The zebrafish (Danio rerio)/tumor xenograft model represents a powerful new model system in cancer. Here, we describe a novel exploitation of the zebrafish model to investigate tumor angiogenesis, a pivotal step in cancer progression and target for antitumor therapies. Human and murine tumor cell lines that express the angiogenic fibroblast growth factor (FGF) 2 and/or vascular endothelial growth factor (VEGF) induce the rapid formation of a new microvasculature when grafted close to the developing subintestinal vessels of zebrafish embryos at 48 h postfertilization. Instead, no angiogenic response was exerted by related cell clones defective in the production of these angiogenic growth factors. The newly formed blood vessels sprout from the subintestinal plexus of the zebrafish embryo, penetrate the tumor graft, and express the transcripts for the zebrafish orthologues of the early endothelial markers Fli-1, VEGF receptor-2 (VEGFR2/KDR), and VE-cadherin. Accordingly, green fluorescent protein-positive neovessels infiltrate the graft when tumor cells are injected in transgenic VEGFR2:G-RCFP zebrafish embryos that express green fluorescent protein under the control of the VEGFR2/KDR promoter. Systemic exposure of zebrafish embryos immediately after tumor cell injection to prototypic antiangiogenic inhibitors, including the FGF receptor tyrosine kinase inhibitor SU5402 and the VEGFR2/KDR tyrosine kinase inhibitor SU5416, suppresses tumor-induced angiogenesis without affecting normal blood vessel development. Accordingly, VE-cadherin gene inactivation by antisense morpholino oligonucleotide injection inhibits tumor neovascularization without affecting the development of intersegmental and subintestinal vessels. These data show that the zebrafish/tumor xenograft model represents a novel tool for investigating the neovascularization process exploitable for drug discovery and gene targeting in tumor angiogenesis. PMID- 17409397 TI - High HSP90 expression is associated with decreased survival in breast cancer. AB - The heat shock protein HSP90 chaperones proteins implicated in breast cancer progression, including Her2/neu. HSP90-targeting agents are in clinical trials for breast cancer. HSP90 expression is high in breast cancer cell lines, yet no large studies have been conducted on expression in human tumors and the association with clinical/pathologic variables. Tissue microarrays containing 10 cell lines and primary specimens from 655 patients with 10-year follow-up were assessed using our automated quantitative analysis (AQUA) method; we used cytokeratin to define pixels as breast cancer (tumor mask) within the array spot and measured HSP90 expression within the mask using Cy5-conjugated antibodies. We similarly assessed estrogen receptor, progesterone receptor, and Her2/neu expression. HSP90 expression was more variable in human tumors than in cell lines (P < 0.0001). High HSP90 expression was associated with decreased survival (P = 0.0024). On multivariable analysis, high HSP90 expression remained an independent prognostic marker. High HSP90 expression was associated with high Her2/neu and estrogen receptor, large tumors, high nuclear grade, and lymph node involvement. Although HSP90 levels were high in all our cell lines, expression in tumors was more variable. High HSP90 expression in primary breast cancer defines a population of patients with decreased survival. Evaluation of HSP90 expression in early-stage breast cancer may identify a subset of patients requiring more aggressive or pathway-targeted treatment. Prospective studies are needed to confirm the prognostic role of HSP90, as well as the predictive role of HSP90 expression in patients treated with HSP90 inhibitors. PMID- 17409398 TI - Materializing the potential of small interfering RNA via a tumor-targeting nanodelivery system. AB - The field of small interfering RNA (siRNA) as potent sequence-selective inhibitors of transcription is rapidly developing. However, until now, low transfection efficiency, poor tissue penetration, and nonspecific immune stimulation by in vivo administered siRNAs have delayed their therapeutic application. Their potential as anticancer therapeutics hinges on the availability of a vehicle that can be systemically administered, safely and repeatedly, and will deliver the siRNA specifically and efficiently to the tumor, both primary tumors and metastases. We have developed a nanosized immunoliposome based delivery complex (scL) that, when systemically administered, will preferentially target and deliver molecules useful in gene medicine, including plasmid DNA and antisense oligonucleotides, to tumor cells wherever they occur in the body. This tumor-targeting nanoparticle delivery vehicle can also deliver siRNA to both primary and metastatic disease. We have also enhanced the efficiency of this complex by the inclusion of a pH-sensitive histidine-lysine peptide in the complex (scL-HoKC) and by delivery of a modified hybrid (DNA-RNA) anti-HER-2 siRNA molecule. Scanning probe microscopy confirms that this modified complex maintains its nanoscale size. More importantly, we show that this nanoimmunoliposome anti-HER-2 siRNA complex can sensitize human tumor cells to chemotherapeutics, silence the target gene and affect its downstream pathway components in vivo, and significantly inhibit tumor growth in a pancreatic cancer model. Thus, this complex has the potential to help translate the potent effects of siRNA into a clinically viable anticancer therapeutic. PMID- 17409399 TI - Two common chromosome 8q24 variants are associated with increased risk for prostate cancer. AB - Two variants (rs1447295/DG8S737) of chromosome 8q24 were recently reported to be associated with increased risk of prostate cancer (PC). To confirm this finding, we genotyped and compared the frequencies of these polymorphisms among 1,121 Caucasian men with PC (435 men with familial PC, 491 men with sporadic PC, and 195 men with aggressive PC) to 545 population-based controls. For the single nucleotide polymorphism marker rs1447295, frequencies of the minor allele (A) were 10.3% in controls, 11.9% in sporadic cases, 16.7% in familial cases, and 17.2% in aggressive cases. Compared with controls, the A allele was significantly more common in both familial PC [odds ratios (OR), 1.93; 95% confidence intervals (95% CI), 1.37-2.72; P = 0.0004] and aggressive PC (OR, 1.87; 95% CI, 1.28-2.74; P = 0.0005) but not for sporadic PC (OR, 1.16; 95% CI, 0.85-1.58; P = 0.25). Although the A allele was more frequent in aggressive PC cases when compared with controls, the allele frequencies were similar among cases with high- and low grade PC (Gleason grades <7 and >/=7, respectively). For the microsatellite marker DG8S737, the -8 allele was significantly more frequent in familial PC (OR, 1.68; 95% CI, 1.09-2.60; P = 0.031), whereas the -10 allele was more frequent in aggressive PC (OR, 2.85; 95% CI, 1.52-5.36; P = 0.0004). Haplotype analysis showed significant differences in haplotype frequencies between the familial PC (P = 0.006) and aggressive PC (P = 0.005) cases versus controls. The -8/A haplotype showed the strongest association with familial PC (P = 0.008), whereas the -10/A haplotype was most strongly associated with aggressive PC (P = 0.00005). These results further confirm the importance of these two polymorphic variants (rs1447295 and DG8S737) as risk factors for PC. However, the mechanism explaining this increased risk has not yet been established. PMID- 17409400 TI - A common 8q24 variant in prostate and breast cancer from a large nested case control study. AB - Two recent studies independently identified polymorphisms in the 8q24 region, including a single nucleotide polymorphism (rs1447295), strongly associated with prostate cancer risk. Here, we replicate the overall association in a large nested case-control study from the National Cancer Institute Breast and Prostate Cancer Cohort Consortium using 6,637 prostate cancer cases and 7,361 matched controls. We also examine whether this polymorphism is associated with breast cancer among 2,604 Caucasian breast cancer cases and 3,118 matched controls. The rs1447295 marker was strongly associated with prostate cancer among Caucasians (P = 1.23 x 10(-13)). When we exclude the Multiethnic Cohort samples, previously reported by Freedman et al., the association remains highly significant (P = 8.64 x 10(-13)). Compared with wild-type homozygotes, carriers with one copy of the minor allele had an OR(AC) = 1.34 (99% confidence intervals, 1.19-1.50) and carriers with two copies of the minor allele had an OR(AA) = 1.86 (99% confidence intervals, 1.30-2.67). Among African Americans, the genotype association was statistically significant in men diagnosed with prostate cancer at an early age (P = 0.011) and nonsignificant for those diagnosed at a later age (P = 0.924). This difference in risk by age at diagnosis was not present among Caucasians. We found no statistically significant difference in risk when tumors were classified by Gleason score, stage, or mortality. We found no association between rs1447295 and breast cancer risk (P = 0.590). Although the gene responsible has yet to be identified, the validation of this marker in this large sample of prostate cancer cases leaves little room for the possibility of a false-positive result. PMID- 17409401 TI - Aurora a and B overexpression and centrosome amplification in early estrogen induced tumor foci in the Syrian hamster kidney: implications for chromosomal instability, aneuploidy, and neoplasia. AB - Estrogen-induced Syrian hamster tumors in the kidney represent a useful model to gain insight into the role of estrogens in oncogenic processes. We provided evidence that early tumor foci in the kidney arise from interstitial ectopic uterine-like germinal stem cells, and that early tumor foci and well-established tumors are highly aneuploid (92-94%). The molecular mechanisms whereby estrogens mediate this process are unclear. Here, we report that estrogen treatment induced significant increases in Aurora A protein expression (8.7-fold), activity (2.6 fold), mRNA (6.0-fold), and Aurora B protein expression (4.6-fold) in tumors, compared with age-matched cholesterol-treated kidneys. Immunohistochemistry revealed that this increase in Aurora A and B protein expression was essentially confined to cells within early and large tumor foci at 3.5 and 6 months of estrogen treatment, respectively. Upon estrogen withdrawal or coadministration of tamoxifen for 10 days, a 78% to 79% and 81% to 64% reduction in Aurora A and B expression, respectively, were observed in primary tumors compared with tumors continuously exposed to estrogens. These data indicate that overexpressed Aurora A and B in these tumors are under estrogen control via estrogen receptor alpha. Aurora A coenriched with the centrosome fraction isolated from tumors in the kidney. Centrosome amplification (number and area/cell) was detected in early tumor foci and large tumors but not in adjacent uninvolved or age-matched control kidneys. Taken together, these data indicate that persistent overexpression of Aurora A and B is under estrogen control, and is coincident with centrosome amplification, chromosomal instability, and aneuploidy, and represent an important mechanism driving tumorigenesis. PMID- 17409402 TI - Fatty acid synthase inhibition activates AMP-activated protein kinase in SKOV3 human ovarian cancer cells. AB - Fatty acid synthase (FAS), the enzyme responsible for the de novo synthesis of fatty acids, is highly expressed in ovarian cancers and most common human carcinomas. Inhibition of FAS and activation of AMP-activated protein kinase (AMPK) have been shown to be cytotoxic to human cancer cells in vitro and in vivo. In this report, we explore the cytotoxic mechanism of action of FAS inhibition and show that C93, a synthetic FAS inhibitor, increases the AMP/ATP ratio, activating AMPK in SKOV3 human ovarian cancer cells, which leads to cytotoxicity. As a physiologic consequence of AMPK activation, acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid synthesis, was phosphorylated and inhibited whereas glucose oxidation was increased. Despite these attempts to conserve energy, the AMP/ATP ratio increased with worsening cellular redox status. Pretreatment of SKOV3 cells with compound C, an AMPK inhibitor, substantially rescued the cells from C93 cytotoxicity, indicating its dependence on AMPK activation. 5-(Tetradecyloxy)-2-furoic acid, an ACC inhibitor, did not activate AMPK despite inhibiting fatty acid synthesis pathway activity and was not significantly cytotoxic to SKOV3 cells. This indicates that substrate accumulation from FAS inhibition triggering AMPK activation, not end-product depletion of fatty acids, is likely responsible for AMPK activation. C93 also exhibited significant antitumor activity and apoptosis against SKOV3 xenografts in athymic mice without significant weight loss or cytotoxicity to proliferating cellular compartments such as bone marrow, gastrointestinal tract, or skin. Thus, pharmacologic FAS inhibition selectively activates AMPK in ovarian cancer cells, inducing cytotoxicity while sparing most normal human tissues from the pleiotropic effects of AMPK activation. PMID- 17409403 TI - Tumor suppression by IFN regulatory factor-1 is mediated by transcriptional down regulation of cyclin D1. AB - IFNs have been ascribed to mediate antitumor effects. IFN regulatory factor-1 (IRF-1) is a major target gene of IFNs. It inhibits cell proliferation and oncogenic transformation. Here, we show that 60% of all mRNAs deregulated by oncogenic transformation mediated by c-myc and H-ras are reverted to the expression levels of nontransformed cells by IRF-1. These include cell cycle regulating genes. An indirect target is cyclin D1. Activation of IRF-1 decreased cyclin D1 expression and cyclin-dependent kinase 4 kinase activity concomitant with change in the levels of hyperphosphorylated retinoblastoma protein. These effects are mediated by inhibition of the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway and a transcriptional repression of cyclin D1. As shown by in vitro assays and tumor growth in nude mice, IRF-1-mediated effects on cell cycle progression were found to be overridden by ectopic expression of cyclin D1. Conversely, decrease of cyclin D1 by RNA interference experiments prevents transformation and tumor growth. The data show that cyclin D1 is a key target for IRF-1-mediated tumor-suppressive effects. PMID- 17409404 TI - Molecular dissection of hyperdiploid multiple myeloma by gene expression profiling. AB - Hyperdiploid multiple myeloma (H-MM) is the most common form of myeloma. In this gene expression profiling study, we show that H-MM is defined by a protein biosynthesis signature that is primarily driven by a gene dosage mechanism as a result of trisomic chromosomes. Within H-MM, four independently validated patient clusters overexpressing nonoverlapping sets of genes that form cognate pathways/networks that have potential biological importance in multiple myeloma were identified. One prominent cluster, cluster 1, is characterized by high expression of cancer testis antigen and proliferation-associated genes. Tumors from these patients were more proliferative than tumors in other clusters (median plasma cell labeling index, 3.8; P < 0.05). Another cluster, cluster 3, is characterized by genes involved in tumor necrosis factor/nuclear factor-kappaB signaling and antiapoptosis. These patients have better response to bortezomib as compared with patients within other clusters (70% versus 29%; P = 0.02). Furthermore, for a group of patients generally thought to have better prognosis, a cluster of patients with short survival (cluster 1; median survival, 27 months) could be identified. This analysis illustrates the heterogeneity within H-MM and the importance of defining specific cytogenetic prognostic factors. Furthermore, the signatures that defined these clusters may provide a basis for tailoring treatment to individual patients. PMID- 17409405 TI - Isogenic normal basal and luminal mammary epithelial isolated by a novel method show a differential response to ionizing radiation. AB - Epithelial cells within the normal breast duct seem to be the primary target for neoplastic transformation events that eventually produce breast cancer. Normal epithelial cells are easily isolated and propagated using standard techniques. However, these techniques almost invariably result in populations of cells that are largely basal in character. Because only approximately 20% of human breast cancers exhibit a basal phenotype, our understanding of the disease may be skewed by using these cells as the primary comparator to cancer. Further, because germ line mutations in BRCA1 yield breast cancers that are most often of the basal type, a comparison of normal basal and luminal cells could yield insight into the tissue and cell type specificity of this hereditary cancer susceptibility gene. In this report, we describe a simplified and efficient method for isolating basal and luminal cells from normal human breast tissue. These isogenic cells can be independently propagated and maintain phenotypic markers consistent with their respective lineages. Using these cultured cells, we show that basal and luminal cells exhibit distinct responses to ionizing radiation. Basal cells undergo a rapid but labile cell cycle arrest, whereas luminal cells show a much more durable arrest, primarily at the G(2)-M boundary. Molecular markers, including p53 protein accumulation, p53-activated genes, and BRCA1 nuclear focus formation all correlate with the respective cell cycle responses. Further, we show that short-term cultures of human breast tissue fragments treated with ionizing radiation show a similar phenomenon as indicated by the biphasic accumulation of p53 protein in the basal versus luminal layer. Together, these results indicate that normal basal cells have a transitory cell cycle arrest after DNA damage that may underlie their increased susceptibility to transformation after the loss of functional BRCA1. PMID- 17409406 TI - Tumor-specific cooperation of retinoblastoma protein family and Snf5 inactivation. AB - Malignant rhabdoid tumors (MRT) are rare aggressive cancers that occur in young children. Seventy-five percent of sporadic MRTs harbor inactivating SNF5 mutations, and mice heterozygous for an Snf5-null allele develop MRTs with partial penetrance. The diagnosis of choroid plexus carcinomas (CPC) in addition to MRTs in families with a single mutant SNF5 allele prompted us to assess the role of SNF5 loss in CPC in genetically engineered mice. With high frequency, TgT(121) mice develop CPCs that are initiated by inactivation of retinoblastoma protein (pRb) and related proteins p107 and p130. However, CPC penetrance and latency were not significantly affected by Snf5 heterozygosity, consistent with recent evidence that CPCs in SNF5 families were, in many cases, misdiagnosed MRTs. Surprisingly, although the CPC phenotype was unaffected, TgT(121);Snf5(+/-) mice developed MRTs with increased penetrance and decreased latency compared with TgT(121);Snf5(+/+) littermates. MRTs expressed the T(121) protein with a concomitant increase in mitotic activity. The predominant appearance of TgT(121);Snf5(+/-) MRTs in the spinal cord led to the discovery that these tumors likely arose from a subset of spinal cord neural progenitor cells expressing T(121) rather than from transdifferentiation of CPC. Significantly, the target cell type(s) for MRT is unknown. Hence, this study not only shows that pRb(f) and SNF5 inactivation cooperate to induce MRTs but also provides new insight into the MRT target population. PMID- 17409407 TI - Hyperthermia activates a subset of ataxia-telangiectasia mutated effectors independent of DNA strand breaks and heat shock protein 70 status. AB - All cells have intricately coupled sensing and signaling mechanisms that regulate the cellular outcome following exposure to genotoxic agents such as ionizing radiation (IR). In the IR-induced signaling pathway, specific protein events, such as ataxia-telangiectasia mutated protein (ATM) activation and histone H2AX phosphorylation (gamma-H2AX), are mechanistically well characterized. How these mechanisms can be altered, especially by clinically relevant agents, is not clear. Here we show that hyperthermia, an effective radiosensitizer, can induce several steps associated with IR signaling in cells. Hyperthermia induces gamma H2AX foci formation similar to foci formed in response to IR exposure, and heat induced gamma-H2AX foci formation is dependent on ATM but independent of heat shock protein 70 expression. Hyperthermia also enhanced ATM kinase activity and increased cellular ATM autophosphorylation. The hyperthermia-induced increase in ATM phosphorylation was independent of Mre11 function. Similar to IR, hyperthermia also induced MDC1 foci formation; however, it did not induce all of the characteristic signals associated with irradiation because formation of 53BP1 and SMC1 foci was not observed in heated cells but occurred in irradiated cells. Additionally, induction of chromosomal DNA strand breaks was observed in IR exposed but not in heated cells. These results indicate that hyperthermia activates signaling pathways that overlap with those activated by IR-induced DNA damage. Moreover, prior activation of ATM or other components of the IR-induced signaling pathway by heat may interfere with the normal IR-induced signaling required for chromosomal DNA double-strand break repair, thus resulting in increased cellular radiosensitivity. PMID- 17409408 TI - Evidence that in xeroderma pigmentosum variant cells, which lack DNA polymerase eta, DNA polymerase iota causes the very high frequency and unique spectrum of UV induced mutations. AB - Xeroderma pigmentosum variant (XPV) patients have normal DNA excision repair, yet are predisposed to develop sunlight-induced cancer. They exhibit a 25-fold higher than normal frequency of UV-induced mutations and very unusual kinds (spectrum), mainly transversions. The primary defect in XPV cells is the lack of functional DNA polymerase (Pol) eta, the translesion synthesis DNA polymerase that readily inserts adenine nucleotides opposite photoproducts involving thymine. The high frequency and striking difference in kinds of UV-induced mutations in XPV cells strongly suggest that, in the absence of Pol eta, an abnormally error-prone polymerase substitutes. In vitro replication studies of Pol iota show that it replicates past 5'T-T3' and 5'T-U3' cyclobutane pyrimidine dimers, incorporating G or T nucleotides opposite the 3' nucleotide. To test the hypothesis that Pol iota causes the high frequency and abnormal spectrum of UV-induced mutations in XPV cells, we identified an unlimited lifespan XPV cell line expressing two forms of Pol iota, whose frequency of UV-induced mutations is twice that of XPV cells expressing one form. We eliminated expression of one form and compared the parental cells and derivatives for the frequency and kinds of UV-induced mutations. All exhibited similar sensitivity to the cytotoxicity of UV((254 nm)), and the kinds of mutations induced were identical, but the frequency of mutations induced in the derivatives was reduced to /=20% CD138(+)HLA-DR(+) cells (median, 50%; range, 23-100). Because primary plasma cells cannot be efficiently cultured in vitro, we used a panel of MM cell lines with a dim/negative to bright HLA-DR expression to evaluate 1D09C3-induced cell death. Annexin V/propidium iodide (PI) staining showed that 1D09C3-induced cell death correlated with constitutive HLA DR expression. Induction of HLA-DR by IFN-gamma restored the sensitivity of HLA DR dim cell lines to 1D09C3. In vivo, the combined IFN-gamma/1D09C3 treatment significantly increased the median survival of nonobese diabetic/severe combined immunodeficient mice xenografted with KMS-11 cell line, compared with controls (147 versus 48 days, P 2-fold higher levels than under normal glucose conditions. Among these genes, asparagine synthetase (ASNS) was studied in detail. Although it is known to be associated with drug resistance in leukemia and oncogenesis triggered by mutated p53, its function is yet to be determined. In this study, we found that glucose deprivation induced the overexpression of ASNS through an AMP-activated protein kinase-independent and activating transcription factor-4-dependent manner and that ASNS protects pancreatic cancer cells from apoptosis induced by glucose deprivation itself. ASNS overexpression also induced resistance to apoptosis triggered by cisplatin [cis-diammine-dichloroplatinum (CDDP)] and carboplatin, but not by 5-fluorouracil, paclitaxel, etoposide, or gemcitabine. We show that glucose deprivation induces the activation of c-jun NH(2)-terminal kinase (JNK)/stress-activated protein kinase (SAPK) in a mock transfectant but not in an ASNS transfectant. Consequently, an inhibitor of JNK/SAPK decreased the sensitivity of pancreatic cancer cells to apoptosis by glucose deprivation and CDDP. These results strongly suggest that ASNS is induced by glucose deprivation and may play a pivotal role in the survival of pancreatic cancer cells under glucose-deprived conditions. PMID- 17409445 TI - Induction of Cdc25B regulates cell cycle resumption after genotoxic stress. AB - Cdc25 phosphatases propel cell cycle progression by activating cyclin-dependent kinases (Cdk). DNA damage is generally thought to inhibit Cdc25 functionality by inducing proteasomal degradation of Cdc25A and phosphorylation-mediated sequestration of Cdc25B and Cdc25C to the cytoplasm. More recently, a critical role for Cdc25B in the resumption of cell cycle progression through mitosis after DNA damage has been identified. In this study, the fate of Cdc25B after mechanistically distinct DNA-damaging agents (etoposide, cisplatin, bleomycin, ionizing irradiation, or UV irradiation) was examined, and surprisingly a rapid increase in cellular Cdc25B levels was observed after DNA damage. Using UV irradiation as the prototypic damaging agent, we found that the increase in Cdc25B levels was checkpoint dependent and was controlled by a p53-independent mechanism. Cdc25B levels controlled the number of cells progressing into mitosis after UV, but they did not affect G(2)-M checkpoint engagement immediately after DNA damage. Increased Cdc25B reduced the time required for cell cycle resumption. These data support a model in which Cdc25B accumulation is an important anticipatory event for cell cycle resumption after DNA damage. PMID- 17409446 TI - 2-Deoxy-D-glucose combined with cisplatin enhances cytotoxicity via metabolic oxidative stress in human head and neck cancer cells. AB - Glucose deprivation has been hypothesized to cause cytotoxicity by inducing metabolic oxidative stress in human cancer cells. The current work tests the hypothesis that 2-deoxy-d-glucose (2DG) combined with cisplatin [cis diamminedichloroplatinum(II)] can enhance cytotoxicity in human head and neck cancer cells (FaDu) by mechanisms involving oxidative stress. Exposure of FaDu cells to the combination of 2DG and cisplatin resulted in a significant decrease in cell survival when compared with 2DG or cisplatin alone. Treatment with 2DG and cisplatin also caused perturbations in parameters indicative of oxidative stress, including decreased intracellular total glutathione and increased percentage of glutathione disulfide. Simultaneous treatment with the thiol antioxidant N-acetylcysteine (NAC) inhibited parameters indicative of oxidative stress, as well as protected FaDu cells from the cytotoxic effects of cisplatin alone and the combination of 2DG and cisplatin. In addition, polyethylene glycol conjugated antioxidant enzymes (PEG-superoxide dismutase and PEG-catalase) also protected FaDu cells from 2DG toxicity. An inhibitor of glutathione synthesis, l buthionine-[S,R]-sulfoximine (BSO), sensitized FaDu cells to the cytotoxic effects of 2DG and cisplatin, and these effects were inhibited by NAC. Furthermore, the combination of 2DG, cisplatin, and BSO significantly increased the percentage of glutathione disulfide, which was also inhibited by NAC. These results support the hypothesis that exposure of human head and neck cancer cells to 2DG combined with cisplatin enhances cytotoxicity via metabolic oxidative stress. These findings provide a strong biochemical rationale for evaluating inhibitors of glucose and hydroperoxide metabolism in combination with cisplatin for the treatment of head and neck cancer. PMID- 17409447 TI - Parthenolide inhibits tubulin carboxypeptidase activity. AB - Microtubules are centrally involved in cell division, being the principal components of mitotic spindle. Tubulin, the constituent of microtubules, can be cyclically modified on its alpha-subunit by enzymatic removal of the COOH terminal tyrosine residue by an ill-defined tubulin carboxypeptidase (TCP) and its readdition by tubulin tyrosine ligase (TTL). We and others have previously shown that suppression of TTL and resulting accumulation of detyrosinated tubulin are frequent in human cancers of poor prognosis. Explanations for the involvement of TTL and detyrosinated tubulin in tumor progression arise from the recent discovery that tubulin detyrosination leads to CAP-Gly protein mislocalization, which correlates with defects in spindle positioning during mitosis. Impaired control of spindle positioning is one factor favoring tumor invasiveness. Thus, TCP could be a target for developing novel therapeutic strategies against advanced stages of cancers. Inhibitors of TCP, by reversing abnormal detyrosinated tubulin accumulation in tumor cells, could impair tumor progression. TCP has never been isolated and this has hampered search of specific inhibitors. In this article, we describe a cell-based assay of TCP activity and its use to screen a library of natural extracts for their inhibitory potency. This led to the isolation of two sesquiterpene lactones. We subsequently found that parthenolide, a structurally related compound, can efficiently inhibit TCP. This inhibitory activity is a new specific property of parthenolide independent of its action on the nuclear factor-kappaB pathway. Parthenolide is also known for its anticancer properties. Thus, TCP inhibition could be one of the underlying mechanisms of these anticancer properties. PMID- 17409448 TI - Cholesterol starvation induces differentiation of human leukemia HL-60 cells. AB - Cholesterol metabolism is particularly active in malignant, proliferative cells, whereas cholesterol starvation has been shown to inhibit cell proliferation. Inhibition of enzymes involved in cholesterol biosynthesis at steps before the formation of 7-dehydrocholesterol has been shown to selectively affect cell cycle progression from G(2) phase in human promyelocytic HL-60 cells. In the present work, we explored whether cholesterol starvation by culture in cholesterol-free medium and treatment with different distal cholesterol biosynthesis inhibitors induces differentiation of HL-60 cells. Treatment with SKF 104976, an inhibitor of lanosterol 14-alpha demethylase, or with zaragozic acid, which inhibits squalene synthase, caused morphologic changes alongside respiratory burst activity and expression of cluster of differentiation antigen 11c (CD11c) but not cluster of differentiation antigen 14. These effects were comparable to those produced by all-trans retinoic acid, which induces HL-60 cells to differentiate following a granulocyte lineage. In contrast, they differed from those produced by vitamin D(3), which promotes monocyte differentiation. The specificity of the response was confirmed by addition of cholesterol to the culture medium. Treatment with PD 98059, an inhibitor of extracellular signal-regulated kinase, abolished both the activation of NADPH oxidase and the expression of the CD11c marker. In sharp contrast, BM 15766, which inhibits sterol Delta(7)-reductase, failed to induce differentiation or arrest cell proliferation. These results show that changes in the sterol composition may trigger a differentiation response and highlight the potential of cholesterol pathway inhibition as a possible tool for use in cancer therapy. PMID- 17409449 TI - Improvement of antitumor activity by gene amplification with a replicating but nondisseminating adenovirus. AB - Gene therapy is a promising approach for cancer treatment; however, efficacy of current vectors remains insufficient. To improve the success of suicide gene therapy, we constructed a replication-competent adenoviral vector that has its protease gene deleted and expresses bacterial cytosine deaminase fused with bacterial uracil phosphoribosyltransferase (CU). The prodrug, 5-fluorocytosine, is transformed into the highly toxic and tissue-diffusible 5-fluorouracil by CU in infected cells. This vector is incapable of producing infectious particles but is able to undergo a single round of replication, thereby increasing transgene copy number and expression. In the presence of 5-FC, compared with the first generation vector (AdCU), the replication-competent vector, Ad(dPS)CU-IRES-E1A, was significantly more efficacious for in vitro tumor cell killing and in bystander assays, whereas 25-fold fewer viral particles were required in a three dimensional spheroid model. For in vivo experiments, in which virus was injected into preestablished intracranial glioma xenografts, followed by 5-FC treatment, mice receiving Ad(dPS)CU-IRES-E1A had significantly smaller tumors at 35 days postinjection as well as significantly longer median survival than mice treated with the replication-deficient, protease-deleted vector [Ad(dPS)CU]. In an immunocompetent syngeneic model, Ad(dPS)CU + 5-FC-treated mice had a median survival of only 23 days, whereas Ad(dPS)CU-IRES-E1A + 5-FC-treated animals had a survival of 57.1% at 365 days. In conclusion, Ad(dPS)CU-IRES-E1A in the presence of 5-FC produces more potent tumoricidal effects than its replication-deficient counterparts. PMID- 17409450 TI - CXCL10 promotes invasion-related properties in human colorectal carcinoma cells. AB - CXCL10 was recently shown to exert antimalignancy functions by influencing the tumor microenvironment. Here, we have taken a different approach, investigating the effects of CXCL10 directly on tumor-promoting functions in colorectal carcinoma (CRC) cells. CXCL10 expression was detected in preferred metastatic sites of CRC (liver, lungs, and lymph nodes), and its CXCR3 receptor was expressed by eight CRC cell lines (detected: reverse transcription-PCR and/or flow cytometry). Detailed analysis was done on two cell lines derived from primary CRC tumors (SW480, KM12C) and their metastatic descendents (SW620 and KM12SM). The three known variants of CXCR3 (CXCR3-A, CXCR3-B, and CXCR3-alt) were detected in all four cell lines. CXCR3 expression was also observed on colorectal tumor cells in biopsies of CRC patients (immunohistochemistry). CXCL10 and CXCR3 expression were potently induced in CRC cells by Interferon gamma and all four CRC cell lines responded to CXCL10 by extracellular signal-regulated kinase 1/2 dephosphorylation. The chemokine did not affect tumor cell growth or angiogenesis related functions in the tumor cells, such as CXCL8 and vascular endothelial growth factor secretion. Importantly, CXCL10 significantly up-regulated invasion related properties in CRC cells: It promoted matrix metalloproteinase 9 expression and induced CRC cell migration. Of note, CXCL10-induced migration was detected only in the two metastatic cells and not in their primary counterparts. Also, CXCL10 promoted the adhesion of metastatic cells to laminin. These results suggest that CXCL10 can be exploited by CRC cells toward their progression, thus possibly antagonizing the antimalignancy effects of the chemokine on the tumor microenvironment. Therefore, care should be taken when considering CXCL10 as a therapeutic antitumor modality for CRC treatment. PMID- 17409451 TI - Active induction of tumor-specific IgE antibodies by oral mimotope vaccination. AB - A role of IgE antibodies in cancer surveillance has been implicated for a long time. Studies dealing with IgE antibodies directly targeted to tumor antigens have shown marked anticancer effects mediated by this antibody class. Thus, the basic function of IgE antibodies may be to control tumor growth. Thus far, cancer specific IgE has only been applied passively. Consequently, the aim of this study was to establish an active vaccination protocol to induce tumor antigen-specific IgE antibodies, and to evaluate functional properties. We previously generated epitope mimics, so-called mimotopes, for the epitope recognized by the anti-HER-2 antibody trastuzumab. Upon i.p. immunizations, IgG antibodies with trastuzumab like properties could be elicited. In the present study, we immunized BALB/c mice via the oral route with these trastuzumab mimotopes, under simultaneous neutralization and suppression of gastric acid. As shown in preceding experiments, this feeding regimen effectively induces Th2 immune responses. Oral immunizations with trastuzumab mimotopes under hypoacidic conditions indeed resulted in the formation of IgE antibodies towards the HER-2 antigen. Moreover, anti-HER-2 IgE-sensitized effector cells mediated SK-BR-3 target cell lysis in an antibody-dependent cytotoxicity assay. We conclude that directed and epitope specific induction of IgE against tumor antigens is feasible with an oral mimotope vaccination regimen, and that these antibodies mediate anticancer effects. PMID- 17409452 TI - Evidences that leptin up-regulates E-cadherin expression in breast cancer: effects on tumor growth and progression. AB - Leptin, a cytokine mainly produced by adipocytes, seems to play a crucial role in mammary carcinogenesis. In the present study, we explored the mechanism of leptin mediated promotion of breast tumor growth using xenograft MCF-7 in 45-day-old female nude mice, and an in vitro model represented by MCF-7 three-dimensional cultures. Xenograft tumors, obtained only in animals with estradiol (E(2)) pellet implants, doubled control value after 13 weeks of leptin exposure. In three dimensional cultures, leptin and/or E(2) enhanced cell-cell adhesion. This increased aggregation seems to be dependent on E-cadherin because it was completely abrogated in the presence of function-blocking E-cadherin antibody or EGTA, a calcium-chelating agent. In three-dimensional cultures, leptin and/or E(2) treatment significantly increased cell growth, which was abrogated when E cadherin function was blocked. These findings well correlated with an increase of mRNA and protein content of E-cadherin in three-dimensional cultures and in xenografts. In MCF-7 cells both hormones were able to activate E-cadherin promoter. Mutagenesis studies, electrophoretic mobility shift assay, and chromatin immunoprecipitation assays revealed that cyclic AMP-responsive element binding protein and Sp1 motifs, present on E-cadherin promoter, were important for the up-regulatory effects induced by both hormones on E-cadherin expression in breast cancer MCF-7 cells. In conclusion, the present study shows how leptin is able to promote tumor cell proliferation and homotypic tumor cell adhesion via an increase of E-cadherin expression. This combined effect may give reasonable emphasis to the important role of this cytokine in stimulating primary breast tumor cell growth and progression, particularly in obese women. PMID- 17409453 TI - Androgen deprivation increases p300 expression in prostate cancer cells. AB - Standard therapy for nonorgan confined prostate cancer aims to block the production or action of androgens. Although initially successful, antiandrogen therapy eventually fails and androgen depletion independent (ADI) disease emerges. Remarkably, ADI prostate cancers still rely on a functional androgen receptor (AR). Aberrant expression of coregulatory proteins required for the formation of productive AR transcriptional complexes is critical for ADI AR activation. Previously, we have shown that the transcriptional coactivator p300 is required for ADI activation of the AR and is up-regulated in prostate cancer, in which its expression is associated with cell proliferation and predicts aggressive tumor features. The mechanism responsible for the deregulated expression of p300, however, remains elusive. Here, we show that p300 expression in prostate cancer cells is subject to androgen regulation. In several prostate cancer model systems, addition of synthetic and natural androgens led to decreased expression of p300 in a time-dependent and dose-dependent manner. Experiments using AR antagonists or small interfering RNA targeting the AR revealed that down-regulation of p300 depends entirely on the presence of a functional AR. It is noteworthy that androgens down-regulated p300 protein expression while leaving messenger levels unaltered. Conversely, both short-term and long-term androgen deprivation resulted in marked up-regulation of p300 expression. The androgen deprivation-induced increase in p300 expression was not affected by the addition of cytokines or growth factors or by cotreatment with antiandrogens. Moreover, increased p300 expression upon androgen starvation is crucial for prostate cancer cell proliferation, as loss of p300 expression severely reduces expression of cyclins governing G(1)-S and G(2)-M cell cycle transition and decreases 5-bromo-2'-deoxyuridine incorporation. PMID- 17409454 TI - Phosphoprotein pathway mapping: Akt/mammalian target of rapamycin activation is negatively associated with childhood rhabdomyosarcoma survival. AB - Mapping of protein signaling networks within tumors can identify new targets for therapy and provide a means to stratify patients for individualized therapy. Despite advances in combination chemotherapy, the overall survival for childhood rhabdomyosarcoma remains approximately 60%. A critical goal is to identify functionally important protein signaling defects associated with treatment failure for the 40% nonresponder cohort. Here, we show, by phosphoproteomic network analysis of microdissected tumor cells, that interlinked components of the Akt/mammalian target of rapamycin (mTOR) pathway exhibited increased levels of phosphorylation for tumors of patients with short-term survival. Specimens (n = 59) were obtained from the Children's Oncology Group Intergroup Rhabdomyosarcoma Study (IRS) IV, D9502 and D9803, with 12-year follow-up. High phosphorylation levels were associated with poor overall and poor disease-free survival: Akt Ser(473) (overall survival P < 0.001, recurrence-free survival P < 0.0009), 4EBP1 Thr(37/46) (overall survival P < 0.0110, recurrence-free survival P < 0.0106), eIF4G Ser(1108) (overall survival P < 0.0017, recurrence-free survival P < 0.0072), and p70S6 Thr(389) (overall survival P < 0.0085, recurrence free survival P < 0.0296). Moreover, the findings support an altered interrelationship between the insulin receptor substrate (IRS-1) and Akt/mTOR pathway proteins (P < 0.0027) for tumors from patients with poor survival. The functional significance of this pathway was tested using CCI-779 in a mouse xenograft model. CCI-779 suppressed phosphorylation of mTOR downstream proteins and greatly reduced the growth of two different rhabdomyosarcoma (RD embryonal P = 0.00008; Rh30 alveolar P = 0.0002) cell lines compared with controls. These results suggest that phosphoprotein mapping of the Akt/mTOR pathway should be studied further as a means to select patients to receive mTOR/IRS pathway inhibitors before administration of chemotherapy. PMID- 17409455 TI - Relation of a hypoxia metagene derived from head and neck cancer to prognosis of multiple cancers. AB - Affymetrix U133plus2 GeneChips were used to profile 59 head and neck squamous cell cancers. A hypoxia metagene was obtained by analysis of genes whose in vivo expression clustered with the expression of 10 well-known hypoxia-regulated genes (e.g., CA9, GLUT1, and VEGF). To minimize random aggregation, strongly correlated up-regulated genes appearing in >50% of clusters defined a signature comprising 99 genes, of which 27% were previously known to be hypoxia associated. The median RNA expression of the 99 genes in the signature was an independent prognostic factor for recurrence-free survival in a publicly available head and neck cancer data set, outdoing the original intrinsic classifier. In a published breast cancer series, the hypoxia signature was a significant prognostic factor for overall survival independent of clinicopathologic risk factors and a trained profile. The work highlights the validity and potential of using data from analysis of in vitro stress pathways for deriving a biological metagene/gene signature in vivo. PMID- 17409456 TI - A high-throughput study in melanoma identifies epithelial-mesenchymal transition as a major determinant of metastasis. AB - Metastatic disease is the primary cause of death in cutaneous malignant melanoma (CMM) patients. To understand the mechanisms of CMM metastasis and identify potential predictive markers, we analyzed gene-expression profiles of 34 vertical growth phase melanoma cases using cDNA microarrays. All patients had a minimum follow-up of 36 months. Twenty-one cases developed nodal metastatic disease and 13 did not. Comparison of gene expression profiling of metastatic and nonmetastatic melanoma cases identified 243 genes with a >2-fold differential expression ratio and a false discovery rate of <0.2 (206 up-regulated and 37 down regulated). This set of genes included molecules involved in cell cycle and apoptosis regulation, epithelial-mesenchymal transition (EMT), signal transduction, nucleic acid binding and transcription, protein synthesis and degradation, metabolism, and a specific group of melanoma- and neural-related proteins. Validation of these expression data in an independent series of melanomas using tissue microarrays confirmed that the expression of a set of proteins included in the EMT group (N-cadherin, osteopontin, and SPARC/osteonectin) were significantly associated with metastasis development. Our results suggest that EMT-related genes contribute to the promotion of the metastatic phenotype in primary CMM by supporting specific adhesive, invasive, and migratory properties. These data give a better understanding of the biology of this aggressive tumor and may provide new prognostic and patient stratification markers in addition to potential therapeutic targets. PMID- 17409457 TI - Autoantibody profiles reveal ubiquilin 1 as a humoral immune response target in lung adenocarcinoma. AB - There is considerable evidence that the presence of cancer can elicit a humoral immune response to specific proteins in the host, and these resulting autoantibodies may have potential as noninvasive biomarkers. To characterize the autoantibody repertoire present in the sera of patients with lung adenocarcinoma, we developed a high-density peptide microarray derived from biopanning a lung cancer phage display library. Using a 2,304-element microarray, we interrogated a total of 250 sera from Michigan lung cancer patients and noncancer controls to develop an "autoantibody profile" of lung adenocarcinoma. A set of 22 discriminating peptides derived from a training set of 125 serum samples from lung adenocarcinoma patients and control subjects was found to predict cancer status with 85% sensitivity and 86% specificity in an independent test set of 125 sera. Sequencing of the immunoreactive phage-peptide clones identified candidate humoral immune response targets in lung adenocarcinoma, including ubiquilin 1, a protein that regulates the degradation of several ubiquitin-dependent proteasome substrates. An independent validation set of 122 serum samples from Pittsburgh was examined using two overlapping clones of ubiquilin 1 that showed 0.79 and 0.74 of the area under the receiver operating characteristics curve, respectively. Significantly increased levels of both ubiquilin 1 mRNA and protein, as well as reduced levels of the phosphorylated form of this protein, were detected in lung tumors. Immunofluorescence using anti-ubiquilin 1 antibodies confirmed intracellular expression within tumors cells. These studies indicate that autoantibody profiles, as well as individual candidates, may be useful for the noninvasive detection of lung adenocarcinoma. PMID- 17409458 TI - Silibinin inhibits inflammatory and angiogenic attributes in photocarcinogenesis in SKH-1 hairless mice. AB - Sunscreens partially filter UVB and, therefore, could partially prevent skin cancer; however, efficient approaches are desired to effectively prevent photocarcinogenesis. It is hypothesized that nontoxic pharmacologically active natural compounds can increase photoprotective effects. Our completed studies suggest that silibinin, a bioactive phytochemical, strongly prevents photocarcinogenesis; however, its mechanism is not fully understood. Herein, for the first time, we used a clinically relevant UVB dose (30 mJ/cm(2)/day) to examine the photoprotective effect and associated mechanisms of silibinin in SKH1 hairless mice. Topical or dietary silibinin treatment caused a strong protection against photocarcinogenesis in terms of delay in tumor appearance, multiplicity, and volume. Analyses of normal skin, uninvolved skin from tumor-bearing mice, and skin tumors showed a statistically significant decrease (P < 0.05-0.001) in inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) levels by silibinin. Concomitantly, phospho-signal transducers and activators of transcription 3 (Tyr(705)) and phospho-p65(Ser(536)) were also decreased by silibinin, which are potential up-stream regulators of iNOS and COX-2. Simultaneously, silibinin also decreased UVB-caused increase in cell proliferation and microvessel density. In tumors, hypoxia-inducible factor 1alpha (HIF-1alpha) and vascular endothelial growth factor protein levels were decreased by silibinin. Further analysis showed that silibinin inhibited UVB-caused phosphorylation and nuclear translocation of STAT3 and p65, as well as nuclear factor kappaB (NF-kappaB) DNA binding activity. Together, these results suggest that silibinin causes a strong protective effect against photocarcinogenesis via down-regulation of inflammatory and angiogenic responses, involving HIF-1alpha, STAT3, and NF-kappaB transcription factors, as well as COX2 and iNOS. PMID- 17409459 TI - Filling time of a lamellar capillary-filling semen analysis chamber is a rapid, precise, and accurate method to assess viscosity of seminal plasma. PMID- 17409460 TI - Acute adverse effects of the indenopyridine CDB-4022 on the ultrastructure of sertoli cells, spermatocytes, and spermatids in rat testes: comparison to the known sertoli cell toxicant Di-n-pentylphthalate (DPP). AB - Acute effects of CDB-4022 on testicular ultrastructure were determined. Rats were treated orally with vehicle or a maximally effective single dose of CDB-4022 or Di-n-pentylphthalate (DPP). Preserved testes were processed for transmission electron microscopy. Sertoli and germ cells of vehicle-treated rats demonstrated normal morphological characteristics. Disruption of Sertoli cell ultrastructure was apparent in CDB-4022-treated rats by 3 hours. A decrease in the presence of nucleoli, an increase in the amount and diameter of swollen smooth endoplasmic reticulum, and decreases in cytoplasmic ground substance were observed. The severity of these degenerative effects increased at 6 and 12 hours: Vacuoles were apparent; increased cellular debris, swollen mitochondria, and phagocytic structures were observed; and membranes became more disorganized. Similar ultrastructural changes were observed in the Sertoli cells of DPP-treated rats. By 3 hours, spermatocytes and spermatids were adversely affected by CDB-4022 treatment with swelling of the nuclear envelope. The Step 8 spermatids were especially noteworthy; chromatin was more diffuse and rarefied, the nuclear envelopes were incomplete or broken, and the position of the spermatid nucleus within the cell and relative to Sertoli cell cytoplasm was unusual. Fusion of spermatids to form giant cells was observed by 12 hours. CDB-4022 acts acutely on Sertoli cells to induce marked cellular rarefaction and degeneration, but not necrosis. A rapid and direct effect of CDB-4022 on spermatocytes and spermatids was observed. The antispermatogenic activity of CDB-4022 appears to be a consequence of direct effects on Sertoli and germ cells. PMID- 17409461 TI - Azoospermia due to a unique de novo balanced reciprocal translocation (Y;1) (q12;q25). PMID- 17409462 TI - Development of normal reference values for seminal reactive oxygen species and their correlation with leukocytes and semen parameters in a fertile population. AB - Although reactive oxygen species (ROSs) are clearly implicated in the pathogenesis of male infertility, few studies have attempted to define the basal levels of ROSs in fertile men. Levels of ROSs are highly influenced by the presence of leukocytes and are associated with decreased seminal parameters. The objective of our study was to determine the normal ROS reference values in neat and washed semen of a fertile population and to correlate the leukocyte concentrations with seminal parameters. We evaluated 114 fertile men seeking vasectomy and 47 subfertile patients as a positive control. All samples were subjected to semen analysis and Endtz testing; chemiluminescence assay was used to determine ROS levels. All seminal parameters were significantly higher in the fertile men than in the subfertile patients. In nonleukocytospermic samples, ROS levels were lower in the fertile men than in the subfertile patients in neat (0.29 [0.18, 0.54] vs 0.94 [0.38, 1.51]) (P = .001) and washed semen (5.73 [1.90, 14.71] vs 23.4 [9.46, 115.55]) (P = .001). Similarly, in samples with leukocytes (Entdz, less than 1 x 10(6)/mL), ROS levels were lower in the fertile men in neat (0.75 [0.27, 1.71] vs 2.0 [0.97, 27.41]) (P = .001) and washed semen (15.85 [4.18, 62.16] vs 239.83 [33.4, 1193.75]) (P < .0001). As expected, samples with leukocytes had significantly higher ROS values in washed and neat semen. In the fertile population, ROSs were positively correlated with leukocytes and negatively correlated with sperm count and motility. In semen samples without leukocytes, the normality cutoff of ROSs was 0.55 x 10(4) counted photons per minute with 76.4% area under the curve (AUC) in the neat samples and 10.0 x 10(4) counted photons per minute with 77% AUC in the washed samples. In semen samples with leukocytes, the cutoff for ROSs in neat samples was 1.25 with 72.7% AUC and 51.5 with 81% AUC in the washed samples. We defined the cutoff levels of ROSs in a fertile population. Seminal leukocyte levels below 1 x 10(6)/mL were associated with increased ROSs. ROS levels were positively correlated with leukocytes and negatively correlated with sperm motility and concentration. Patients with normal seminal parameters and lower seminal leukocyte levels may benefit from therapeutic interventions that improve semen quality. PMID- 17409463 TI - A retrospective comparison of pregnancy outcome following conventional oocyte insemination vs intracytoplasmic sperm injection for isolated abnormalities in sperm morphology using strict criteria. AB - Initial in vitro fertilization (IVF)-embryo transfer studies found poor fertilization and pregnancy rates following conventional fertilization of oocytes when using sperm with <4% normal morphology using strict criteria. Some consider today that sperm with only < or =5% normal morphology using strict criteria are associated with infertility. However, other studies have disputed the diagnostic potential of low strict morphology in identifying subnormal male fertility. Based on the original studies most IVF centers perform intracytoplasmic sperm injection (ICSI) when the sperm shows low morphology using strict criteria to allow selection of normal sperm. However, ICSI adds extra time for the embryologist and extra expense for the infertile couple. The present study retrospectively compared fertilization, pregnancy, and implantation rates according to the 2 methods of oocyte fertilization with sperm having normal morphology using strict criteria of < or =5% in women < or =39 years. All fresh embryo transfers were performed on day 3. There was a significantly higher fertilization rate with ICSI. However, there were significantly higher clinical and delivered pregnancy and implantation rates following conventional insemination. The rate of canceled transfers due to no available embryo was 1.9% with conventional insemination vs 1.5% for ICSI in women with adequate egg reserve. Hopefully, this retrospective study will generate interest in a prospective study. PMID- 17409464 TI - Tag STS in the AZF region associated with azoospermia in a Tunisian population. AB - The aim of this study was to determine the appropriate tag sequence tagged site (STS) associated with azoospermia. We evaluated the incidence of Y chromosome microdeletions in Tunisian infertile male patients by polymerase chain reaction using 14 STSs in the azoospermia factor (AZF) region of Yq11. A logistic regression analysis was performed to test the association of STSs with semen quality. Haploview version 3.11 was used to identify the possible blocks of deletion involving a minimum number of STSs and that can be used to tag the deletion block in future analysis. Using the 14 STSs, 48% infertile patients (102 of 210) had microdeletions of Y chromosome but, following the European Academy of Andrology guidelines, only 16% of patients had microdeletions. A statistically significant difference was found with some STSs for azoospermia and oligozoospermia. A candidate combinaison composed of 4 STSs (RBMY-sy157-sy84 sy130) was associated with azoospermia in a Tunisian population. According to this study, this tag can be used in the screening of Y chromosome microdeletions before assisted reproduction treatment in a Tunisian population. PMID- 17409465 TI - Isolation and characterization of LNCaP sublines differing in hormone sensitivity. AB - Prostate cancer is a heterogeneous disease with varying degrees of androgen sensitivity. In this study, we performed a limiting dilution of human prostate LNCaP cells, and isolated two sublines, LNCaP-E9 and LNCaP-G4, with differential hormone-sensitivity. Two LNCaP sublines were obtained by the limiting dilution method. The growth of E9 cells was decreased in the presence of androgens, while that of androgen-treated G4 cells was biphasic. Although the androgen receptor expression level in E9 cells was similar to that seen in G4 cells, the expression of PSA mRNA and protein was significantly lower in the E9 cells. Moreover, the androgen-based stimulation of PSA mRNA expression was less sensitive in E9 cells than G4 cells. Intracellular zinc level did not differ between E9 and G4 cells, but ZnT3 mRNA expression was significantly higher in the E9 cells. When the cells were grafted at the subrenal capsule, the number of CD31-positive vessels with a lumen was approximately 2.5 times higher than that in G4 tumors. LNCaP-E9 cells show lower androgen sensitivity than LNCaP-G4 cells. E9 and G4 cells would be helpful for understanding the biology of hormone-refractory prostate cancer. PMID- 17409466 TI - Microvillar size and espin expression in principal cells of the adult rat epididymis are regulated by androgens. AB - Principal cells of the epididymis are the most prominent cell type and are noted for an apical cell surface studded with microvilli. The latter contain channel proteins that condition the microenvironment of epididymal lumen and promote sperm maturation; however, the regulation of the structure and integrity of microvilli is not well known. Espins are a family of proteins implicated in microvillar growth. The objectives of this study were to assess the regulation of espin in epididymal principal cells both in vitro and in vivo. Treatment of immortalized rat caput epididymal (RCE) cells with increasing doses of a homogenized testicular extract revealed a dose-dependent increase in the size of microvilli. Reverse transcriptase-polymerase chain reaction (RT-PCR) of adult rat epididymal RNA using espin-specific primers indicated the presence of a band at about 290 base pairs (bp) in all regions. Western blot analysis using affinity purified espin antibody confirmed the presence of an approximately 110-kDa band in the epididymis, corresponding to espin isoform 1. In adult rats, immunocytochemistry revealed espin expression over principal cells. In orchidectomized rats, espin expression was significantly reduced, whereas ligation of the efferent ducts resulted in a decrease of espin expression but not to the extent of orchidectomy. The fact that espin expression was restored to control levels in orchidectomized rats supplemented with high levels of testosterone indicated that its expression was dependent on androgens and not on other lumicrine factors derived from the testis. Taken together, these data indicate that espin is expressed in the epididymis and is regulated by androgens. PMID- 17409467 TI - Chordee without hypospadias: report of 79 Chinese prepubertal patients. AB - A series of Chinese prepubertal patients with congenital chordee without hypospadias is presented and the clinical data described. From July 1999 to September 2006, 79 boys with congenital chordee without hypospadias were treated in the Department of Pediatric Surgery, West China Hospital of Sichuan University, China. The ages ranged from 21 months to 14 years, with a mean of 76.8 months (6.4 years). The patients were categorized according to structural defect into 4 groups, with the aid of intraoperative artificial erection. Group I included those with skin tethering (28 cases, 35.4%); group II, fascial chordee (22, 27.8%); group III, corporal disproportion (10, 12.7%); and group IV, urethral tethering (19, 24.1%). Chordee-related structural defect was considered the only criterion for classification, and urethral dysgenesis influenced the choice of surgical procedure. The chordee in group I patients was corrected with penile degloving; group II, release of dense fibrous tissue in addition; group III, dorsal-midline-plication-based correction; and group IV, longitudinal-island flap-urethroplasty-based repair. At a mean follow-up of 14.8 months (range, 2 to 63), all patients had penile straightening except 1 group III patient with residual curvature that was managed upon reoperation. Glans dehiscence occurred in 1 group II patient who underwent a tubularized incised plate urethroplasty. Urethrocutaneous fistula and urethral stricture were found in 2 group IV patients who underwent island flap urethroplasty. With the categorization based on structural defect, chordee without hypospadias may be managed well with minimized complications. PMID- 17409468 TI - Effects of the chemotherapeutic agents for non-Hodgkin lymphoma, cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), on the male rat reproductive system and progeny outcome. AB - Chemotherapy of non-Hodgkin lymphoma (NHL) with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) is associated with significant gonadal damage. Our goal was to determine the impact of CHOP chemotherapy on the male reproductive system, fertility, and progeny outcome in the rat model. Adult male Sprague-Dawley rats received saline or CHOP, 4 cycles of 3 weeks each, at doses analogous to 1/3x, 2/3x, or 1x the human dose; males were mated to evaluate effects on progeny outcome. Reproductive organ weights were significantly decreased in the 1x CHOP-exposed group. The spermatozoal contents of the testes and epididymides were decreased in 1x CHOP-treated males; the 1/3x and 2/3x doses also affected testicular sperm contents. Seminiferous tubule diameters were decreased by 20% in 1x CHOP-treated males. Damage ranged from the presence of small vacuoles in the epithelium to tubules deprived of spermatocytes and spermatids and was accompanied by an increased incidence of germ cell apoptosis. The acridine orange assay revealed a significant increase in sperm with abnormal DNA integrity profiles in the 1x CHOP group. Despite effects on germ cell number and quality, CHOP-exposed rats remained fertile. However, a 50% decrease in live fetuses was observed in litters sired by 1x CHOP-treated males due to a significant increase in both pre-implantation and postimplantation losses; postimplantation loss was also elevated among litters sired by 2/3x CHOP-treated males. Thus, CHOP treatment affected both the quantity and quality of male germ cells; conceptal loss is a sensitive measure of the integrity of the male genome. PMID- 17409469 TI - Minocycline increases quality and longevity of chronic neural recordings. AB - Brain/machine interfaces could potentially be used in the treatment of a host of neurological disorders ranging from paralysis to sensory deficits. Insertion of chronic micro-electrode arrays into neural tissue initiates a host of immunological responses, which typically leads to the formation of a cellular sheath around the implant, resulting in the loss of useful signals. Minocycline has been shown to have neuroprotective and neurorestorative effects in certain neural injury and neurodegenerative disease models. This study examined the effects of minocycline administration on the quality and longevity of chronic multi-channel microwire neural implants 1 week and 1 month post-implantation in auditory cortex. The mean signal-to-noise ratio for the minocycline group stabilized at the end of week 1 and remained above 4.6 throughout the following 3 weeks. The control group signal-to-noise ratio dropped throughout the duration of the study and at the end of 4 weeks was 2.6. Furthermore, 68% of electrodes from the minocycline group showed significant stimulus-driven activity at week 4 compared to 12.5% of electrodes in the control group. There was a significant reduction in the number of activated astrocytes around the implant in minocycline subjects, as well as a reduction in total area occupied by activated astrocytes at 1 and 4 weeks. PMID- 17409470 TI - Toward closed-loop optimization of deep brain stimulation for Parkinson's disease: concepts and lessons from a computational model. AB - Deep brain stimulation (DBS) of the subthalamic nucleus with periodic, high frequency pulse trains is an increasingly standard therapy for advanced Parkinson's disease. Here, we propose that a closed-loop global optimization algorithm may identify novel DBS waveforms that could be more effective than their high-frequency counterparts. We use results from a computational model of the Parkinsonian basal ganglia to illustrate general issues relevant to eventual clinical or experimental tests of such an algorithm. Specifically, while the relationship between DBS characteristics and performance is highly complex, global search methods appear able to identify novel and effective waveforms with convergence rates that are acceptably fast to merit further investigation in laboratory or clinical settings. PMID- 17409471 TI - Electrochemical polymerization of conducting polymers in living neural tissue. AB - A number of biomedical devices require extended electrical communication with surrounding tissue. Significant improvements in device performance would be achieved if it were possible to maintain communication with target cells despite the reactive, insulating scar tissue that forms at the device-tissue interface. Here, we report that the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) can be polymerized directly within living neural tissue resulting in an electrically conductive network that is integrated within the tissue. Nano and microscale PEDOT filaments extend out from electrode sites, presumably forming within extracellular spaces. The cloud of PEDOT filaments penetrates out into the tissue far enough that it should be possible to bypass fibrous scar tissue and contact surrounding healthy neurons. These electrically functional, diffuse conducting polymer networks grown directly within tissue signify a new paradigm for creating soft, low impedance implantable electrodes. PMID- 17409472 TI - A review of classification algorithms for EEG-based brain-computer interfaces. AB - In this paper we review classification algorithms used to design brain-computer interface (BCI) systems based on electroencephalography (EEG). We briefly present the commonly employed algorithms and describe their critical properties. Based on the literature, we compare them in terms of performance and provide guidelines to choose the suitable classification algorithm(s) for a specific BCI. PMID- 17409473 TI - Prosthetic interfaces with the visual system: biological issues. AB - The design of effective visual prostheses for the blind represents a challenge for biomedical engineers and neuroscientists. Significant progress has been made in the miniaturization and processing power of prosthesis electronics; however development lags in the design and construction of effective machine-brain interfaces with visual system neurons. This review summarizes what has been learned about stimulating neurons in the human and primate retina, lateral geniculate nucleus and visual cortex. Each level of the visual system presents unique challenges for neural interface design. Blind patients with the retinal degenerative disease retinitis pigmentosa (RP) are a common population in clinical trials of visual prostheses. The visual performance abilities of normals and RP patients are compared. To generate pattern vision in blind patients, the visual prosthetic interface must effectively stimulate the retinotopically organized neurons in the central visual field to elicit patterned visual percepts. The development of more biologically compatible methods of stimulating visual system neurons is critical to the development of finer spatial percepts. Prosthesis electrode arrays need to adapt to different optimal stimulus locations, stimulus patterns, and patient disease states. PMID- 17409474 TI - A survey of signal processing algorithms in brain-computer interfaces based on electrical brain signals. AB - Brain-computer interfaces (BCIs) aim at providing a non-muscular channel for sending commands to the external world using the electroencephalographic activity or other electrophysiological measures of the brain function. An essential factor in the successful operation of BCI systems is the methods used to process the brain signals. In the BCI literature, however, there is no comprehensive review of the signal processing techniques used. This work presents the first such comprehensive survey of all BCI designs using electrical signal recordings published prior to January 2006. Detailed results from this survey are presented and discussed. The following key research questions are addressed: (1) what are the key signal processing components of a BCI, (2) what signal processing algorithms have been used in BCIs and (3) which signal processing techniques have received more attention? PMID- 17409475 TI - Suppression of axonal conduction by sinusoidal stimulation in rat hippocampus in vitro. AB - Deep brain stimulation (DBS), also known as high frequency stimulation (HFS), is a well-established therapy for Parkinson's disease and essential tremor, and shows promise for the therapeutic control of epilepsy. However, the direct effect of DBS on neural elements close to the stimulating electrode remains an important unanswered question. Computational studies have suggested that HFS has a dual effect on neural elements inhibiting cell bodies, while exciting axons. Prior experiments have shown that sinusoidal HFS (50 Hz) can suppress synaptic and non synaptic cellular activity in several in vitro epilepsy models, in all layers of the hippocampus. However, the effects of HFS on axons near the electrode are still unclear. In the present study, we tested the hypothesis that HFS suppresses axonal conduction in vitro. Sinusoidal HFS was applied to the alvear axon field of transverse rat hippocampal slices. The results show that HFS suppresses the alvear compound action potential (CAP) as well as the CA1 antidromic evoked potential (AEP). Complete suppression was observed as a 100% reduction in the amplitude of the evoked field potential for the duration of the stimulus. Evoked potential width and latency were not significantly affected by sinusoidal HFS. Suppression was dependent on HFS amplitude and frequency, but independent of stimulus duration and synaptic transmission. The frequency dependence of sinusoidal HFS is similar to that observed in clinical DBS, with maximal suppression between 50 and 200 Hz. HFS produced not only suppression of axonal conduction but also a correlated rise in extracellular potassium. These data provide new insights into the effects of HFS on neuronal elements, and show that HFS can block axonal activity through non-synaptic mechanisms. PMID- 17409476 TI - Classification of motor imagery by means of cortical current density estimation and Von Neumann entropy. AB - The goal of the present study is to employ the source imaging methods such as cortical current density estimation for the classification of left- and right hand motor imagery tasks, which may be used for brain-computer interface (BCI) applications. The scalp recorded EEG was first preprocessed by surface Laplacian filtering, time-frequency filtering, noise normalization and independent component analysis. Then the cortical imaging technique was used to solve the EEG inverse problem. Cortical current density distributions of left and right trials were classified from each other by exploiting the concept of Von Neumann entropy. The proposed method was tested on three human subjects (180 trials each) and a maximum accuracy of 91.5% and an average accuracy of 88% were obtained. The present results confirm the hypothesis that source analysis methods may improve accuracy for classification of motor imagery tasks. The present promising results using source analysis for classification of motor imagery enhances our ability of performing source analysis from single trial EEG data recorded on the scalp, and may have applications to improved BCI systems. PMID- 17409477 TI - Neurite outgrowth and branching of PC12 cells on very soft substrates sharply decreases below a threshold of substrate rigidity. AB - Rationally designed matrices for nerve tissue engineering and encapsulated cell therapies critically rely on a comprehensive understanding of neural response to biochemical as well as biophysical cues. Whereas biochemical cues are established mediators of neuronal behavior (e.g., outgrowth), physical cues such as substrate stiffness have only recently been recognized to influence cell behavior. In this work, we examine the response of PC12 neurites to substrate stiffness. We quantified and controlled fibronectin density on the substrates and measured multiple neurite behaviors (e.g., growth, branching, neurites per cell, per cent cells expressing neurites) in a large sample population. We found that PC12 neurons display a threshold response to substrate stiffness. On the softest substrates tested (shear modulus approximately 10 Pa), neurites were relatively few, short in length and unbranched. On stiffer substrates (shear modulus approximately 10(2)-10(4) Pa), neurites were longer and more branched and a greater percentage of cells expressed neurites; significant differences in these measures were not found on substrates with a shear modulus >10(2) Pa. Based on these data and comparisons with published neurobiology and neuroengineering reports of neurite mechanotransduction, we hypothesize that results from studies of neuronal response to compliant substrates are cell-type dependent and sensitive to ligand density, sample size and the range of stiffness investigated. PMID- 17409478 TI - Interaction of embryonic cortical neurons on nanofibrous scaffolds for neural tissue engineering. AB - The interaction of murine embryonic cortical neurons on randomly orientated electrospun scaffolds of poly(L-lactide) (P(L)LA) and poly(lactide-co-glycolide) (PLGA) is investigated in this study. The scaffolds were surface treated with different concentrations of KOH to partially hydrolyze the surface and therefore change the surface tension. Hydrophilicity did not significantly influence the number of primary and secondary branches; however, it had a considerable effect on neurite extension. For scaffolds with surface tensions of 40-47 dyn cm(-1) there was a significantly greater overall neurite length for both the primary and secondary branches compared with more hydrophilic scaffolds. Another major finding of this work was that the interfibre distance influenced how the neurites extended. When the interfibre distance was greater than approximately 15 microm the neurites followed the fibres and avoided regions of very high fibre density. At interfibre distances less than approximately 15 microm, the neurites traversed between the fibres. Therefore, this study provided little evidence that contact guidance was the dominating cue in directing neurite extension, instead inferring that chemical cues, possibly from adjacent neurons had induced directional change. PMID- 17409479 TI - Thin microelectrodes reduce GFAP expression in the implant site in rodent somatosensory cortex. AB - The objective of this study was to test the hypothesis that neural implants with reduced cross-sectional areas will have less glial scarring associated with implantation injury in long-term experiments. In this study, we implanted nine adult rats with two different implants of 12 microm (n = 6), and 25 microm (n = 6) diameters (cross-sectional areas of 68 microm(2), 232 microm(2) respectively) and the expression of glial fibrilliary acidic protein (GFAP) was assessed after 2 weeks and 4 weeks of implantation. In order to facilitate implantation, the 12 microm diameter implants were coated with poly-glycolic acid (PGA), a biodegradable polymer that degraded within minutes of implantation. In n = 3 animals, 25 microm diameter implants also coated with PGA were implanted and assessed for GFAP expression at the end of 4 weeks of implantation. Statistical analysis of the GFAP expression around the different implants demonstrated that after 2 weeks of implantation there is no statistically significant difference in GFAP expression between the 12 microm and the 25 microm diameter implants. However, after 4 weeks of implantation the implant site of 12 microm diameter implants exhibited a statistically significant reduction in GFAP expression when compared to the implant sites of the 25 microm diameter implants (both with and without the PGA coating). We conclude that in neural implants that are tethered to the skull, implant cross-sectional areas of 68 microm(2) and smaller could lead to a reduced glial scarring under chronic conditions. Future studies with longer implant durations can confirm if this observation remains consistent beyond 4 weeks. PMID- 17409480 TI - Conditional firing probabilities in cultured neuronal networks: a stable underlying structure in widely varying spontaneous activity patterns. AB - To properly observe induced connectivity changes after training sessions, one needs a network model that describes individual relationships in sufficient detail to enable observation of induced changes and yet reveals some kind of stability in these relationships. We analyzed spontaneous firing activity in dissociated rat cortical networks cultured on multi-electrode arrays by means of the conditional firing probability. For all pairs (i, j) of the 60 electrodes, we calculated conditional firing probability (CFP(i,j)[tau]) as the probability of an action potential at electrode j at t = tau, given that one was detected at electrode i at t = 0. If a CFP(i,j)[tau] distribution clearly deviated from a flat one, electrodes i and j were considered to be related. For all related electrode pairs, a function was fitted to the CFP-curve to obtain parameters for 'strength' and 'delay' (i.e. maximum and latency of the maximum of the curve) of each relationship. In young cultures the set of identified relationships changed rather quickly. At 16 days in vitro (DIV) 50% of the set changed within 2 days. Beyond 25 DIV this set stabilized: during a week more than 50% of the set remained intact. Most individual relationships developed rather gradually. Moreover, beyond 25 DIV relational strength appeared quite stable, with coefficients of variation (100 x SD/mean) around 25% in periods of approximately 10 h. CFP analysis provides a robust method to describe the underlying probabilistic structure of highly varying spontaneous activity in cultured cortical networks. It may offer a suitable basis for plasticity studies, in the case of changes in the probabilistic structure. CFP analysis monitors all pairs of electrodes instead of just a selected one. Still, it is likely to describe the network in sufficient detail to detect subtle changes in individual relationships. PMID- 17409481 TI - Fast wave propagation in auditory cortex of an awake cat using a chronic microelectrode array. AB - We investigated fast wave propagation in auditory cortex of an alert cat using a chronically implanted microelectrode array. A custom, real-time imaging template exhibited wave dynamics within the 33-microwire array (3 mm(2)) during ten recording sessions spanning 1 month post implant. Images were based on the spatial arrangement of peri-stimulus time histograms at each recording site in response to auditory stimuli consisting of tone pips between 1 and 10 kHz at 75 dB SPL. Functional images portray stimulus-locked spiking activity and exhibit waves of excitation and inhibition that evolve during the onset, sustained and offset period of the tones. In response to 5 kHz, for example, peak excitation occurred at 27 ms after onset and again at 15 ms following tone offset. Variability of the position of the centroid of excitation during ten recording sessions reached a minimum at 31 ms post onset (sigma = 125 microm) and 18 ms post offset (sigma = 145 microm), suggesting a fine place/time representation of the stimulus in the cortex. The dynamics of these fast waves also depended on stimulus frequency, likely reflecting the tonotopicity in auditory cortex projected from the cochlea. Peak wave velocities of 0.2 m s(-1) were also consistent with those purported across horizontal layers of cat visual cortex. The fine resolution offered by microimaging may be critical for delivering optimal coding strategies used with an auditory prosthesis. Based on the initial results, future studies seek to determine the relevance of these waves to sensory perception and behavior. PMID- 17409482 TI - The influence of electrolyte composition on the in vitro charge-injection limits of activated iridium oxide (AIROF) stimulation electrodes. AB - The effects of ionic conductivity and buffer concentration of electrolytes used for in vitro measurement of the charge-injection limits of activated iridium oxide (AIROF) neural stimulation electrodes have been investigated. Charge injection limits of AIROF microelectrodes were measured in saline with a range of phosphate buffer concentrations from [PO(4)(3-)] = 0 to [PO(4)(3-)] = 103 mM and ionic conductivities from 2-28 mS cm(-1). The charge-injection limits were insensitive to the buffer concentration, but varied significantly with ionic conductivity. Using 0.4 ms cathodal current pulses at 50 Hz, the charge-injection limit increased from 0.5 mC cm(-2) to 2.1 mC cm(-2) as the conductivity was increased from 2 mS cm(-1) to 28 mS cm(-1). An explanation is proposed in which the observed dependence on ionic conductivity arises from non-uniform reduction and oxidation within the porous AIROF and from uncorrected iR-drops that result in an overestimation of the redox potential during pulsing. Conversely, slow sweep-rate cyclic voltammograms (CVs) were sensitive to buffer concentration with the potentials of the primary Ir(3+)/Ir(4+) reduction and oxidation reactions shifting approximately 300 mV as the buffer concentration decreased from [PO(4)(3 )] = 103 mM to [PO(4)(3-)] = 0 mM. The CV response was insensitive to ionic conductivity. A comparison of in vitro AIROF charge-injection limits in commonly employed electrolyte models of extracellular fluid revealed a significant dependence on the electrolyte, with more than a factor of 4 difference under some pulsing conditions, emphasizing the need to select an electrolyte model that closely matches the conductivity and ionic composition of the in vivo environment. PMID- 17409483 TI - Impact of cysts during radiofrequency lesioning in deep brain structures--a simulation and in vitro study. AB - Radiofrequency lesioning of nuclei in the thalamus or the basal ganglia can be used to reduce symptoms caused by e.g. movement disorders such as Parkinson's disease. Enlarged cavities containing cerebrospinal fluid (CSF) are commonly present in the basal ganglia and tend to increase in size and number with age. Since the cavities have different electrical and thermal properties compared with brain tissue, it is likely that they can affect the lesioning process and thereby the treatment outcome. Computer simulations using the finite element method and in vitro experiments have been used to investigate the impact of cysts on lesions' size and shape. Simulations of the electric current and temperature distributions as well as convective movements have been conducted for various sizes, shapes and locations of the cysts as well as different target temperatures. Circulation of the CSF caused by the heating was found to spread heat effectively and the higher electric conductivity of the CSF increased heating of the cyst. These two effects were together able to greatly alter the resulting lesion size and shape when the cyst was in contact with the electrode tip. Similar results were obtained for the experiments. PMID- 17409484 TI - An electronic device for artefact suppression in human local field potential recordings during deep brain stimulation. AB - The clinical efficacy of high-frequency deep brain stimulation (DBS) for Parkinson's disease and other neuropsychiatric disorders likely depends on the modulation of neuronal rhythms in the target nuclei. This modulation could be effectively measured with local field potential (LFP) recordings during DBS. However, a technical drawback that prevents LFPs from being recorded from the DBS target nuclei during stimulation is the stimulus artefact. To solve this problem, we designed and developed 'FilterDBS', an electronic amplification system for artefact-free LFP recordings (in the frequency range 2-40 Hz) during DBS. After defining the estimated system requirements for LFP amplification and DBS artefact suppression, we tested the FilterDBS system by conducting experiments in vitro and in vivo in patients with advanced Parkinson's disease undergoing DBS of the subthalamic nucleus (STN). Under both experimental conditions, in vitro and in vivo, the FilterDBS system completely suppressed the DBS artefact without inducing significant spectral distortion. The FilterDBS device pioneers the development of an adaptive DBS system retroacted by LFPs and can be used in novel closed-loop brain-machine interface applications in patients with neurological disorders. PMID- 17409485 TI - Assessing the direct effects of deep brain stimulation using embedded axon models. AB - To better understand the spatial extent of the direct effects of deep brain stimulation (DBS) on neurons, we implemented a geometrically realistic finite element electrical model incorporating anisotropic and inhomogenous conductivities. The model included the subthalamic nucleus (STN), substantia nigra (SN), zona incerta (ZI), fields of Forel H2 (FF), internal capsule (IC) and Medtronic 3387/3389 electrode. To quantify the effects of stimulation, we extended previous studies by using multi-compartment axon models with geometry and orientation consistent with anatomical features of the brain regions of interest. Simulation of axonal firing produced a map of relative changes in axonal activation. Voltage-controlled stimulation, with clinically typical parameters at the dorso-lateral STN, caused axon activation up to 4 mm from the target. This activation occurred within the FF, IC, SN and ZI with current intensities close to the average injected during DBS (3 mA). A sensitivity analysis of model parameters (fiber size, fiber orientation, degree of inhomogeneity, degree of anisotropy, electrode configuration) revealed that the FF and IC were consistently activated. Direct activation of axons outside the STN suggests that other brain regions may be involved in the beneficial effects of DBS when treating Parkinsonian symptoms. PMID- 17409486 TI - Amplitude and phase coupling measures for feature extraction in an EEG-based brain-computer interface. AB - Most of the feature extraction methods in existing brain-computer interfaces (BCIs) are based on the dynamic behavior of separate signals, without using the coupling information between different brain regions. In this paper, amplitude and phase coupling measures, quantified by a nonlinear regressive coefficient and phase locking value respectively, were used for feature extraction. The two measures were based on three different coupling methods determined by neurophysiological a priori knowledge, and applied to a small number of electrodes of interest, leading to six feature vectors for classification. Five subjects participated in an online BCI experiment during which they were asked to imagine a movement of either the left or right hand. The electroencephalographic (EEG) recordings from all subjects were analyzed offline. The averaged classification accuracies of the five subjects ranged from 87.4% to 92.9% for the six feature vectors and the best classification accuracies of the six feature vectors ranged between 84.4% and 99.6% for the five subjects. The performance of coupling features was compared with that of the autoregressive (AR) feature. Results indicated that coupling measures are appropriate methods for feature extraction in BCIs. Furthermore, the combination of coupling and AR feature can effectively improve the classification accuracy due to their complementarities. PMID- 17409488 TI - Identification of arm movements using correlation of electrocorticographic spectral components and kinematic recordings. AB - The purpose of this study was to explore the possibility of using electrocorticographic (ECoG) recordings from subdural electrodes placed over the motor cortex to identify the upper limb motion performed by a human subject. More specifically, we were trying to identify features in the ECoG signals that could help us determine the type of movement performed by an individual. Two subjects who had subdural electrodes implanted over the motor cortex were asked to perform various motor tasks with the upper limb contralateral to the site of electrode implantation. ECoG signals and upper limb kinematics were recorded while the participants were performing the movements. ECoG frequency components were identified that correlated well with the performed movements measured along 6D coordinates (X, Y, Z, roll, yaw and pitch). These frequencies were grouped using histograms. The resulting histograms had consistent and unique shapes that were representative of individual upper limb movements performed by the participants. Thus, it was possible to identify which movement was performed by the participant without prior knowledge of the arm and hand kinematics. To confirm these findings, a nearest neighbour classifier was applied to identify the specific movement that each participant had performed. The achieved classification accuracy was 89%. PMID- 17409487 TI - Trophic factor induction of human umbilical cord blood cells in vitro and in vivo. AB - The mononuclear fraction of human umbilical cord blood (HUCBmnf) is a mixed cell population that multiple research groups have shown contains cells that can express neural proteins. In these studies, we have examined the ability of the HUCBmnf to express neural antigens after in vitro exposure to defined media supplemented with a cocktail of growth and neurotrophic factors. It is our hypothesis that by treating the HUCBmnf with these developmentally-relevant factors, we can expand the population, enhance the expression of neural antigens and increase cell survival upon transplantation. Prior to growth factor treatment in culture, expression of stem cell antigens is greater in the non-adherent HUCBmnf cells compared to the adherent cells (p < 0.05). Furthermore, treatment of the non-adherent cells with growth factors, increases BrdU incorporation, especially after 14 days in vitro (DIV). In HUCBmnf-embryonic mouse striata co culture, a small number of growth factor treated HUCBmnf cells were able to integrate into the growing neural network and express immature (nestin and TuJ1) and mature (GFAP and MAP2) neural markers. Treated HUCBmnf cells implanted in the subventricular zone predominantly expressed GFAP although some grafted HUCBmnf cells were MAP2 positive. While short-term treatment of HUCBmnf cells with growth and neurotrophic factors enhanced proliferative capacity in vitro and survival of the cells in vivo, the treatment regimen employed was not enough to ensure long term survival of HUCBmnf-derived neurons necessary for cell replacement therapies for neurodegenerative diseases. PMID- 17409489 TI - Microfluidic engineered high cell density three-dimensional neural cultures. AB - Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 microm) 3D neural cultures supported by passive diffusion, cell densities 90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue engineered systems reconstituting 3D cell-cell/cell-matrix interactions. PMID- 17409490 TI - HPLC determination and pharmacokinetic study of homoeriodictyol-7-O-beta-D glucopyranoside in rat plasma and tissues. AB - Homoeriodictyol-7-O-beta-D-glucopyranoside (HEDT-Glu) was isolated from Viscum coloratum and identified by MS, 1H- and 13C-NMR. A HPLC method was developed for determination of HEDT-Glu in rat plasma and tissues. All biological samples were pretreated by protein precipitation with acetone. Vanillin was selected as internal standard. The mobile phase consisted of methanol-water-glacial acetic acid (45 : 55 : 0.5, v/v/v). Good linearity were observed over the concentration ranges of 0.1-200.0 microg.ml(-1) in rat plasma and 0.05-5.0 microg.ml(-1) in tissues. Both intra- and inter-day precisions of HEDT-Glu, expressed as the relative standard deviation, were less than 13.1%. Accuracy, expressed as the relative error, ranged from -0.8 to 5.4% in plasma and from -5.6 to 9.4% in tissues. The mean extraction recovery of HEDT-Glu was above 73.17% in biological samples. The described assay method was successfully applied to the pre-clinical pharmacokinetic study of HEDT-Glu. After intravenous administration of HEDT-Glu to rat, AUC and CL(tot) were 16.04+/-3.19 microg.h.ml(-1) and 0.85+/-0.17 l.kg( 1).h(-1), respectively. T(1/2,alpha) and t(1/2,beta) were 0.06+/-0.01 h and 1.27+/-0.31 h, respectively. HEDT-Glu was cleared from the blood and mainly distributed to the liver and small intestine. PMID- 17409491 TI - Characterization of the molecular clock in mouse peritoneal macrophages. AB - Macrophages play essential roles in the innate immune system. In this study, we show that macrophage functions such as phagocytosis and cytokine/chemokine expressions display a circadian rhythm that is regulated by a molecular clock. Phagocytosis, a crucial early reaction by which macrophages protect their host against foreign particles, exhibited a circadian variation that peaks during the light period and bottoms during the dark period. These diurnal changes of phagocytosis activity in macrophages were induced without exogenous stimulants such as bacterial infection. The expression of the clock genes including brain and muscle Arnt-like protein-1 (BMAL1) exhibited robust circadian rhythms in macrophages. The expression patterns of the clock genes in macrophages were similar to those in the suprachiasmatic nucleus and other peripheral tissues. Among inflammation factors examined, the level of monocyte chemoattractant protein-1 (MCP-1/JE) mRNA exhibited most robust circadian oscillation. Expression of other cytokines such as IL-1beta, IL-6 and TNFalpha showed mild diurnal changes. Knockdown of the BMAL1 expression resulted in a decrease of the MCP-1/JE mRNA level in macrophages. BMAL1 increased significantly but weakly MCP-1/JE promoter activity. MCP-1/JE promoter activity is known to be regulated by nuclear factor-kappa B (NF-kappaB). NF-kappaB activity in BMAL1 knockdown macrophages was lower than that in control cells. Consequently, the circadian expression of MCP 1/JE in macrophages is regulated by BMAL1 through the activation of NF-kappaB. The results obtained in this study indicate that the innate immunoreactions involving macrophages are at least partly regulated by the autonomous clock machinery. PMID- 17409492 TI - Triglyceride accumulation by peroxisome proliferators in rat hepatocytes. AB - Peroxisome proliferators (PxPs) induce peroxisomal beta-oxidation (Px-ox) in the liver of rodents and have a hypolipidemic function. To investigate hypolipidemic effect of PxPs, the relationship between TG fluctuation and Px-ox activity, as an indicator of the function of PxPs, was studied in primary cultured rat hepatocytes. Nafenopin (Nf) treatment of hepatocytes caused an increase in Px-ox activity in association with cellular TG accumulation in a time-dependent manner with a coefficient of r=0.918. This relationship between the activity and cellular TG were obtained using structurally diverse PxPs with a correlation coefficient of r=0.747. Treatment of the hypolipidemic drug, but non-PxP Pravastatin, decreased TG in the medium, but did not have the effects on cellular TG and Px-ox activity. The total amount of TG and diacylglycerol acyltransferase activity, the last enzyme in the TG de novo synthesis pathway, were not affected by Nf treatment. When hepatocytes were cultured with Brefeldin A, cellular TG was accumulated, the same as with Nf, however, Px-ox activity was not enhanced. Nf treatment markedly decreased the level of apolipoprotein B (apo B) in very low density lipoprotein (VLDL) fractions prepared from conditioned media and increased that of cellular apoB by Western blot analysis. Microsomal triglyceride transfer protein activity was not influenced by Nf. Together, with regards to TG lowering effect of PxPs, it is suggested that PxPs cause hepatocellular accumulation of TG without effects on TG biosynthesis and VLDL construction, and they might have inhibitory effect on VLDL secretion process. PMID- 17409493 TI - VEGF-C and VEGF-A synergistically enhance lymph node metastasis of gastric cancer. AB - PURPOSE: Vascular endothelial growth factor C (VEGF-C) is the most important molecule in lymphangiogenesis and its relationship with lymph node metastasis has attracted considerable interest. We investigated the relationship of VEGF-C or VEGF-A with clinicopathological factors in gastric cancer patients. METHODS: Eighty gastric cancer patients who underwent gastric resection were analyzed immunohistochemically for expression of VEGF-C and VEGF-A protein. RESULTS: Positive immunoreactivity of VEGF-C and VEGF-A was observed in 75 (93.8%) and 41 (51.3%) patients, respectively. VEGF-A expression was significantly correlated with tumor differentiation (p=0.0017) and vascular invasion (p=0.0004). And positive relationship of VEGF-C expression was only demonstrated with tumor differentiation (p=0.0168). Interestingly, however, the frequency of lymph node metastasis was significantly higher in the patients with expression of both VEGF C and VEGF-A (strong positive expression, p=0.036). Furthermore, the expression of both was also significantly correlated with depth of tumor invasion, tumor differentiation, lymphatic invasion, and vascular invasion. CONCLUSION: The present results suggest that strong expression of VEGF-A in addition to VEGF-C expression is essential in lymph node metastasis, presumably because enhanced metastatic potential including lymphangiogenesis induced by both VEGF-A and VEGF C is vital in lymph node metastasis of gastric cancer. PMID- 17409494 TI - A novel cyclophane compound, CPPy, facilitates NGF-induced TrkA signal transduction and induces cell differentiation in neuroblastoma. AB - Neuroblastoma (NB) often causes spontaneously regression, and can mature to ganglioneuroma. The form with the most favorable prognosis expresses high levels of TrkA, a high-affinity receptor for nerve growth factor (NGF), whereas advanced NB and associated cell lines have abnormalities in the NGF/TrkA signaling pathway. A novel cyclophane, cyclophane pyridine (CPPy), was designed to conserve the tyrosine phosphorylation of TrkA, thereby enhancing NGF/TrkA signal transduction. We investigated whether this compound improved NGF-induced tyrosine phosphorylation of the Y490 domain of TrkA and conserved the expression of an early gene (c-fos) in human NB cell lines (IMR-32 and NB-39). As determined by Western blotting, TrkA (Y490) phosphorylation was enhanced by the combination of CPPy (10(-8) M) and NGF (100 ng/ml) compared with NGF alone. CPPy also conserved NGF-induced c-fos mRNA expression. Moreover, CPPy induced the morphological differentiation of NB cells, leading to expression of the neuronal marker gene GAP-43. These data suggest that CPPy can induce the differentiation of NB cell lines by facilitating NGF-induced TrkA/Ras/MAPK signal transduction, and may therefore be an effective therapeutic agent for NB. PMID- 17409495 TI - Gatifloxacin induces augmented insulin release and intracellular insulin depletion of pancreatic islet cells. AB - Many hypoglycemic and hyperglycemic episodes associated with clinical use of gatifloxacin (GFLX), a novel fluoroquinolone antimicrobial agent, have been reported in recent years. Some have reported hypoglycemia induced by fluoroquinolones, indicating that these agents may stimulate insulin secretion from pancreatic islet cells. In this study, we investigated the effect of GFLX on insulin homeostasis in islet cells using the insulin secreting cell line, HIT T15. After 1 h incubation with over 100 microM of GFLX, insulin secretion from the cells was significantly augmented. However, the augmentation of insulin release induced by GFLX subsequently reached a plateau. Coincidentally, cellular insulin was decreased by 120 h incubation, and reactivity to re-stimulation by sulfonylurea was suppressed. The GFLX insulin depletion effect was stronger than the effects produced by such other fluoroquinolones as levofloxacin and ciprofloxacin. This study suggests that GFLX should induce insulin oversecretion from pancreatic islet cells in the short-term, and decrease insulin productivity or increase insulin disintegration in the long-term. These results are consistent with the clinical results of GFLX finding that hypoglycemic episodes were seen after a first single administration, and most hyperglycemic episodes were seen more than 2 d after the start of administration. PMID- 17409496 TI - Interaction of Mat-8 (FXYD-3) with Na+/K+-ATPase in colorectal cancer cells. AB - Mat-8 was fused with a Myc-tag or green fluorescent protein at its carboxyl terminus, and then expressed in Chinese hamster ovary K1 cells. Determination of the cellular localization of the tagged proteins suggested that they were localized on the intracellular membrane, being not only detected around the nuclear envelope but also partly overlapping with markers for endosomes and Golgi bodies. However, Mat-8 with the Myc-tag was detected on the plasma membrane as well as the intracellular membrane, when it was expressed in colorectal cancer cells. The membrane fraction of the cancer cells was solubilized and immuno precipitated with an antibody for the Myc-tag. Western-blotting analysis demonstrated that the Na+/K+-ATPase alpha subunit was present in the precipitate. Furthermore, the immuno-precipitate obtained with an antibody for the Na+/K+ ATPase alpha subunit reacted with that for the Myc-tag. These results suggested that Mat-8 could be associated with Na+/K+-ATPase similar to other FXYD family members. The Gly41-->Arg mutation in the transmembrane region of Mat-8 inhibited its association with the Na+/K+-ATPase alpha subunit and localization on the plasma membrane, whereas the Cys44-->Ala or Cys49-->Ala substitution did not. Thus the conserved Gly41 residue in the transmembrane domain could be indispensable for localization of Mat-8 on the cell surface. PMID- 17409497 TI - Increased expression of Lewis X and Y antigens on the cell surface and FUT 4 mRNA during granzyme B-induced Jurkat cell apoptosis. AB - Cytotoxic T cells and natural killer cells play key roles in cell-mediated cytotoxicity and can induce apoptosis in virus-infected and malignant cells by releasing cytotoxic granules. In the current study, apoptosis was induced in Jurkat cells, a human T cell line, by delivering granzyme B into the cells using BioPORTER, a cationic lipid formulation. During granzyme B-induced apoptosis, there was an increase in the cell surface expression of Lewis X and Y antigens. To clarify the roles of initiator and executioner caspases in the expression of Lewis X and Y antigens, we treated Jurkat cells with granzyme B in the presence of caspase 3, 8, and 9 inhibitors. The results indicated that delivery of granzyme B into Jurkat cells induces apoptosis by activating caspase 3 and that caspase 3 but not caspase 8 and 9 plays a key role in enhancing the expression of Lewis X and Y antigens. Real-time PCR revealed that expression of the mRNAs for alpha1,3-fucosyltransferases FUT4 was increased at 3 h during granzyme B-induced apoptosis, while FUT9 mRNA expression gradually increased after 12 h. This increased expression of FUT4 mRNA occurred downstream of caspase 3 activation and resulted in the increased cell surface expression of Lewis X and Y antigens. PMID- 17409498 TI - Antioxidant and vascular protective activities of Cratoxylum formosum, Syzygium gratum and Limnophila aromatica. AB - Phytochemicals contained in dietary plants provide a variety of health benefits and may reduce the risk of cardiovascular diseases. The aqueous extracts from three popular Thai dietary and herbal plants, Cratoxylum formosum, Syzygium gratum, and Limnophila aromatica, were investigated for the antioxidant and vascular protective activities in the in vitro and in vivo models. The free radical scavenging and antioxidant activities of plant extracts were evaluated in vitro by the 1,1-diphenyl-2-picrylhydrazyl assay, the ferric reducing antioxidant power assay, the intracellular antioxidant activity in rat peritoneal macrophages by dihydrofluorescein assay, and the inhibition of nitric oxide (NO) production in RAW 264.7 macrophages. In an animal model of oxidative stress and vascular dysfunction, male Sprague-Dawley rats were orally administered with aqueous plant extracts (1 g/kg/d) or N-acetylcysteine (NAC; 300 mg/kg/d) as a control for 6 d. On day four, all animals except the normal control group, were administered with phenylhydrazine (PHZ) intraperitoneally. It was demonstrated that the plant extracts possessed high free radical scavenging and antioxidant activities. PHZ induced severe hemolysis and hemodynamic disturbances and treatment with the extracts and NAC significantly improved the hemodynamic status. Vascular responsiveness to bradykinin, acetylcholine, and phenylephrine in PHZ-control rats was markedly impaired, and the plant extracts or NAC largely restored the vascular responses. Moreover, the plant extracts prevented loss of blood reduced glutathione and suppressed formation of plasma malondialdehyde, plasma NO metabolites and blood superoxide anion. It was concluded that the plant extracts possess antioxidants and have potential roles in protection of vascular dysfunction. PMID- 17409499 TI - Effect of pH on 1H-NMR spectroscopy of mouse urine. AB - 1H-NMR spectroscopy of urine has been proved to be a powerful tool in metabonomic investigations in the field of chemical toxicity evaluation and disease diagnosis. However, most studies on urinary metabolite profiling by 1H-NMR have been conducted using rat urine. In the course of experiment on 1H-NMR analysis of urine samples of mice administered selenium compound, we noticed a substantial variation of chemical shift of citrate among individual mice. To clarify the effect of urinary pH on chemical shift of citrate, we compared 1H-NMR spectra of urine samples obtained from selenium-treated mice and untreated mice as well as those from untreated rats. The results clearly showed a linear relationship between urinary pH and chemical shift of citrate in 1H-NMR spectra both in mice and rats. The urine of mice exhibited a wider variation of pH, resulting in a wider variation of chemical shift of citrate than that of rats. We also recognized a clear peak of trimethylamine in urine of mice, but not that of rats. These data indicate that more attention should be paid to the characteristics of mouse urine with special reference to pH and trimethylamine metabolism in the analysis of NMR spectroscopy. PMID- 17409500 TI - Extracellular ATP is involved in the induction of apoptosis in murine hematopoietic cells. AB - Extracellular nucleotides have multiple biological actions in processes such as proliferation, differentiation, chemotaxis, and cytokine secretion through P2X receptors on the cell surface. To determine the biological activity of adenosine triphosphate (ATP) and the expression of P2 nucleotide receptors in murine bone marrow-derived hematopoietic cells and stem cells/progenitor cells, we investigated the effects of ATP in assays of cell proliferation and cell death in vitro. Our results demonstrated that several subtypes of P2X receptors were expressed on hematopoietic cells and that P2X7, in particular, was partially expressed in hematopoietic stem cells/progenitor cells. In addition, stimulation of hematopoietic cells with high concentrations of ATP caused severe inhibition of cell proliferation despite the presence of cytokine stimulation. We analyzed the apoptotic effects of stimulation with several different dosages of ATP and confirmed the enhanced apoptotic activity in hematopoietic cells and progenitor cells. Antagonists, against P2X receptors and ATP, suramin and oxidized ATP, inhibited the induction of cell death for murine hematopoietic cells. Our data suggest that extracellular nucleotides may provide a novel and powerful tool for regulating the cell fate of hematopoietic stem cells. PMID- 17409501 TI - Combination of amino acids reduces pigmentation in B16F0 melanoma cells. AB - Amino acids, the building blocks of proteins, play significant roles in numerous physiological events in mammals. As the effects of amino acids on melanogenesis have yet to be demonstrated, the present study was conducted to identify whether amino acids, in particular alanine, glycine, isoleucine and leucine, influence melanogenesis in B16F0 melanoma cells. Glycine and L-isoleucine, but not D isoleucine, reduced melanogenesis in a concentration-dependent manner without any morphological changes in B16F0 melanoma cells. L-Alanine and L-leucine, but not D alanine and D-leucine, also reduced melanogenesis without any morphological changes in B16F0 melanoma cells. However these amino acids did not show a concentration-dependency. Combination of L-alanine and the other amino acids, particularly 4 amino acids combination, had an additive effect on the inhibition of melanogenesis compared with single treatment of L-alanine. None of the amino acids affected the activity of tyrosinase, a key enzyme in melanogenesis. These results suggest that L-alanine, glycine, L-isoleucine and L-leucine, but not the D-form amino acids, have a hypopigmenting effect in B16F0 melanoma cells, and that these effects are not due to the inhibition of tyrosinase activity. Combination of these 4 amino acids had the additive effect on hypopigmentation that was as similar as that of kojic acid. PMID- 17409502 TI - Quercetin-induced PC12 cell death accompanied by caspase-mediated DNA fragmentation. AB - Flavonoids have been reported to be potent antioxidants and beneficial in oxidative stress related diseases. Quercetin, a major flavonoid in food, deserves much attention because of its antioxidative activity. However, the actions of flavonoids including quercetin are complex and paradoxical. Quercetin caused apoptosis and/or cell death in various cells including cancer cells and normal cells. In this study, we investigated the effects of quercetin with or without hydrogen peroxide (H2O2) on cell death of PC12 cells, a neuronal cell line. We showed that quercetin at 10-30 microM alone caused cell death accompanied by caspase-mediated DNA fragmentation in undifferentiated PC12 cells. Quercetin did not inhibit and rather enhanced 0.1 mM H2O2-induced cell death. The toxic effect of quercetin was not inhibited by antioxidants such as N-acetylcysteine and GSH, although H2O2-induced cell death was inhibited by the antioxidants. Quercetin induced cell death was reduced by 2 h treatment with nerve growth factor and serum. In addition, quercetin caused cell death in differentiated PC12 cells that were cultured with nerve growth factor for 6 d. Genistein, a soy isoflavone that has the pro-apoptotic activity, also caused cell death with DNA fragmentation. Further evaluation of the potential of dietary flavonoids as neuroprotective reagents is needed. PMID- 17409503 TI - Gamma-aminobutyric acid specifically inhibits progression of tubular fibrosis and atrophy in nephrectomized rats. AB - Gamma-aminobutyric acid (GABA) was administered orally to rats for 60 d after excision of five-sixths of their kidney volume. A decrease in renal function parameters was observed in these nephrectomized rats. However, the administration of GABA ameliorated renal dysfunction, and a longer administration period of GABA increased its protective effect. In addition, tubular fibrosis was markedly increased at 10 and 60 d in nephrectomized control rats, while GABA administration for 10 d reduced tubular fibrosis to the normal level. Tubular atrophy was markedly induced by nephrectomy, and was significantly reduced by the administration of GABA at 60 d. Furthermore, the nephrectomized control rats exhibited an increased expression level of transforming growth factor-beta1, where GABA significantly decreased it after administration for 10 d. The expression of fibronectin in the tubuli of rats administrated GABA for 60 d was completely and dose-dependently reduced as compared with nephrectomized control rats. However, the improvement effects in glomeruli were less. We also found that GABAA and GABAB receptors were specifically localized in tubuli. Specific agonists for GABAA and GABAB receptors improved renal function. These results suggest that GABA may have a beneficial effect on renal function in nephrectomized rats by inhibiting fibrosis and atrophy primarily in tubuli, and that it ameliorates losses of renal function in renal failure. PMID- 17409504 TI - Fructose 1,6-diphosphate alleviates UV-induced oxidative skin damage in hairless mice. AB - Reactive oxygen species (ROS) are involved in the deleterious effects of UV light on skin. The antioxidant defense system is considered to be crucial for protecting skin from ROS. Recently, we showed that fructose 1,6-diphosphate (FDP), a glycolytic metabolite, reduced oxidative stress in UVB-irradiated keratinocytes. This study set out to determine whether topically applied FDP could exert protective effects against UV-induced skin damage in hairless mice. An in vitro skin permeation study using Franz-type diffusion cells showed that the amount of [14C]-FDP that diffused through the skin increased in a time dependent manner, and about 3.5% of the applied FDP penetrated the skin after 24 h. Topical application of FDP (1%) preserved the endogenous antioxidant capacity of skin such as catalase and glutathione, which were significantly reduced after UVB irradiation without FDP. FDP also reversed the loss of catalase protein and prevented the accumulation of carbonylated proteins induced by UVB irradiation. These results provide evidence that topically administered FDP could penetrate into the skin and attenuate UVB-induced oxidative skin damage in hairless mice. PMID- 17409505 TI - Nilvadipine prevents the impairment of spatial memory induced by cerebral ischemia combined with beta-amyloid in rats. AB - In the present study, we examined the effects of nilvadipine and amlodipine, both dihydropyridine-derivative calcium antagonists, on the impairment of spatial memory induced by a combination of ischemia and beta-amyloid (Abeta). Nilvadipine (3.2 mg/kg, i.p.) significantly prevented the impairment of spatial memory and neuronal apoptosis in this model. By contrast, amlodipine had no effect on this impairment of spatial memory. These findings suggest that nilvadipine may prevent impairment of spatial memory by inhibiting neuronal apoptosis; this drug might therefore be useful for the prevention of the progression to dementia in Alzheimer's disease (AD). PMID- 17409506 TI - Pharmacokinetic study of triptolide, a constituent of immunosuppressive chinese herb medicine, in rats. AB - Triptolide is a potential anti-immune agent, and has shown multi-organic toxicity, however its toxic mechanism remained undiscovered. This paper aimed at characterizing the pharmacokinetic profiles of triptolide in rats to provide the clue to approach the toxic mechanism. The absorption, distribution, metabolism and excretion of triptolide were investigated in male Sprague-Dawley rats after single doses of oral and i.v. administration. After oral administration of 0.6, 1.2 and 2.4 mg/kg, the concentration of triptolide in plasma reached the maximum within 15 min, and declined rapidly with an elimination half-life from 16.81 to 21.70 min. The triptolide kinetics was fitted into one-compartment model after i.v. administration. Oral absolute bioavailability was 72.08% at the dose of 0.6 mg/kg. Triptolide was also rapidly distributed and eliminated in all selected tissues. Less than 1% triptolide of the dose was recovered from the bile, urine or feces as parent drug within 48 h. While triptolide could not be detected in tissues and plasma at 4 h post dose, rats in the group C (oral: 1.2 mg/kg) and D (oral: 2.4 mg/kg) showed obvious toxic response to triptolide and some of rats even died out. It was indicated that triptolide was metabolized extensively, eliminated rapidly, and also showed that the toxicity produced by the triptolide was lag behind the exposure concentration. PMID- 17409507 TI - Effect of Tremella fuciformis on the neurite outgrowth of PC12h cells and the improvement of memory in rats. AB - We investigated the neuritogenic effects of Tremella fuciformis (TF), which has been valued in traditional Chinese medicine as a remedy with nutritive and tonic actions, on PC12h cells. The cognitive improving effects of TF on scopolamine induced (2 mg/kg, s.c.) amnesia in rats were also evaluated with using the Morris water maze task and by performing choline acetyltransferase (ChAT) immunohistochemistry. The water extract of TF (0.01-1 microg/ml) promoted neurite outgrowth of the PC12h cells in a dose dependent manner. TF was highly efficient at the concentration range of 0.1-1 microg/ml. Oral daily treatment with TF (100 or 400 mg/kg) for 14 consecutive days significantly reversed the scopolamine induced deficit in learning and memory, and it alleviated decrease in cholinergic immunoreactivity induced by scopolamine in the medial septum and hippocampus. The results demonstrate that the promotion of neuritogenesis in neuronal culture cells by TF water extract is related with its activity for improving the performance of rats on a spatial learning and memory task. Moreover, the impairments of spatial learning and memory may be attributable to the decrease in activation of the septohippocampal cholinergic system and that TF ameliorated learning and memory deficits partly through its increasing the central cholinergic activity. Therefore, TF could represent a potentially useful agent that is able to improve the function of impaired cognitive processes. PMID- 17409508 TI - Synthesis and biological evaluation of 3-(substituted-benzylidene)-1,3-dihydro indolin derivatives as human protein kinase CK2 and p60(c-Src) tyrosine kinase inhibitors. AB - Human protein kinase CK2 is an ubiquitous serine/threonine kinase that is typically found in tetrameric complexes consisting of two catalytic (alpha and/or alpha') and two regulatory beta subunits. Although there is growing evidence that besides the participation of CK2 in a complex series of cellular functions, this protein kinase is involved in cell viability, cell proliferation, and neoplastic transformation. In the present study, a series of 3-(substituted-benzylidene)-1,3 dihydro-indolin-2-thione derivatives and the corresponding indolin-2-one congeners were tested for their inhibition of human recombinant protein kinase CK2 in vitro. The efficacy of these compounds was compared with their inhibitory results of p60(c-Src) tyrosine kinase. It was found that 3-(substituted benzylidene)-1,3-dihydro-indolin-2-thione derivatives are more effective than indolin-2-one congeners for the inhibition of CK2 and p60(c-Src) tyrosine kinase. PMID- 17409509 TI - Inhibition of melanogenesis by selina-4(14),7(11)-dien-8-one isolated from Atractylodis Rhizoma Alba. AB - To develop effective skin-lightening agents, we tested medicinal herbal extracts for their melanogenic-inhibitory activities. We isolated a sesquiterpenoid compound from the extract of Atractylodis Rhizoma Alba using the bioactivity guided fractionation and identified it as selina-4(14),7(11)-dien-8-one (compound 1) with spectroscopic methods. Compound 1 dramatically reduced melanin synthesis of melan-a cells without any apparent cytotoxicity. Compound 1 did not inhibit cell-free tyrosinase activity but decreased tyrosinase activity in melanocytes. These effects were attributed to reduced expression of melanogenic enzymes such as tyrosinase, tyrosinase-related protein 1 (TRP-1), and tyrosinase-related protein 2 (TRP-2). These results suggest that compound 1 may be an effective skin lightening agent that regulates expression of melanogenic enzymes. PMID- 17409510 TI - Study on the hydroxyl radical scavenging activity changes of ginseng and ginsenoside-Rb2 by heat processing. AB - The free radical scavenging activities of Panax ginseng C.A. MEYER are known to increase by heat processing. Phenolic acids and Maillard reaction products (MRPs) have been suggested as active free radical scavenging components from our previous research, but heat processing-induced chemical and activity changes of ginsenosides considering the Maillard reaction have not yet been fully elucidated. In this study, we investigated the hydroxyl radical (.OH) scavenging activity changes of ginsengs and ginsenoside-Rb2 (Rb2)) by heat processing using an electron spin resonance spectrometer. Especially, Rb2 was heat processed with the same amount of glycine, a frequently used amino acid in the Maillard reaction model system. As a result, the .OH scavenging activities and brown compound levels of ginseng and glycine-Rb2 mixture were increased by heat processing. However, the increase in .OH scavenging activities were not in accordance with the extents of browning. On the other hand, less-polar ginsenosides such as Rg3, Rg5, and Rk1 were generated from the glycine-Rb2 mixture by heat processing. The sugar moieties at carbon-20 of Rb2 were separated by the steaming process, less polar ginsenosides were produced, and then the separated sugar moieties were thought to form MRPs with glycine. From the .OH scavenging activity tests of Rb2, glycine, less-polar ginsenosides, and maltol, the increase in .OH scavenging activity was thought to be more closely related to the generation of .OH scavenging ginsenosides such as 20(S)-Rg3 and Rg5 by heat processing than MRPs. PMID- 17409511 TI - Unilateral lung-selective gene transfer following the administration of naked plasmid DNA onto the pulmonary pleural surface in mice. AB - The purpose of the present study was to examine unilateral lung-selective gene transfer following the administration of naked plasmid DNA (pDNA) onto the pulmonary pleural surface in mice. Naked pDNA was administered intravenously, intraperitoneally, and instilled onto the right pulmonary pleural surface. Four hours later, right pulmonary pleural surface instillation of naked pDNA resulted in high gene expression in the right lung. On the contrary, intravenous and intraperitoneal administration of naked pDNA resulted in no detectable gene expression. After instilling naked pDNA onto the right or left pulmonary pleural surface, gene expressions in the applied lung were significantly higher than those in the other lung and tissues. In addition, gene expressions were detected only in the intrathoracic tissues, not in the intraperitoneal tissues. Four hours after instillation of naked pDNA onto the right pulmonary pleural surface, gene expression in the right lung was the highest, and thereafter gene expression in the right lung decreased gradually. This novel gene transfer method is expected to be a safe and effective treatment against serious lung diseases. PMID- 17409512 TI - Pharmaceutical approach to HIV protease inhibitor atazanavir for bioavailability enhancement based on solid dispersion system. AB - Atazanavir (ATV) is a low oral bioavailability (BA) compound and, clinically, is generally coadministrated with ritonavir (RTV), which boosts the oral BA of ATV by inhibiting cytochrome P450 (CYP) 3A, and P-glycoprotein (Pgp) via the same metabolic pathway. However, depending on pharmacokinetic interaction, RTV-boosted ATV has great potential for other comedication. In this study we demonstrated the pharmaceutical approach to BA improvement of ATV without RTV in rats, based on the solid dispersion system using sodium lauryl sulfate (SLS) as a carrier and Gelucire 50/13 as an absorption enhancer. ATV solid dispersions in SLS were prepared by a conventional solvent method and, at ratios of ATV to SLS of 1 : 2 and 1 : 3, were demonstrated to form an amorphous state in powder X-ray diffraction (PXRD) analysis and exhibited 2.26- and 2.36-fold improvement in a dissolution test in comparison to bulk ATV, respectively. After oral administration to rats, ATV solid dispersion in SLS at a ratio of 1 : 2 showed a 3.5-fold increase in BA compared with bulk ATV. Moreover, the addition of Gelucire 50/13 to ATV solid dispersion, at a total ratio of Gelucire 50/13, ATV and SLS 1 : 1 : 2 gave 7.0- and 4.7-fold increase in Cmax and BA compared with bulk ATV, respectively, when the relative BA to RTV-boosted ATV reached 93%. The results in this study proved that a pharmaceutical approach could improve the bioavailability of ATV without pharmacokinetic interaction with RTV. PMID- 17409513 TI - Involvement of bile salt export pump in flutamide-induced cholestatic hepatitis. AB - The non-steroidal antiandrogen flutamide is widely used for treatment of prostatic cancer, but causes side effects, including cholestatic hepatitis and fulminant hepatitis. We investigated the pathogenesis of flutamide-induced cholestatic hepatitis, focusing on the bile salt export pump (BSEP; ABCB11), which exports bile salts to the bile. We examined the inhibitory effects of flutamide and its active metabolite, hydroxyflutamide, on the transport of taurocholic acid (TCA) by membrane vesicles derived from hBSEP-expressing Sf9 cells. Flutamide inhibited the transport of TCA by hBSEP (IC50 value, about 50 microM), while hydroxyflutamide had no effect at up to 100 microM. When flutamide was administered to rats as a single oral dose of 100 mg/kg, the biliary excretion rate of bolus-injected [3H]TCA was decreased and the liver tissue concentration of flutamide exceeded 50 microM. Repeated doses of flutamide for 5 d (10 mg/kg/d) also decreased the biliary excretion rate of bolus-injected [3H]TCA. In this case, the liver tissue concentration of flutamide was below 0.1 microM. In both cases, no change in the mRNA level of rat Bsep was detected by RT PCR. These results suggest that flutamide itself, but not its major metabolite, may cause cholestasis by inhibiting BSEP-mediated bile salt excretion. PMID- 17409514 TI - Effects of acute renal failure and ganciclovir on the pharmacodynamics of levofloxacin-induced seizures in rats. AB - Seizures have been reported in patients receiving fluoroquinolones, including levofloxacin (LVFX). In the present study, we investigated the effects of experimental renal failure and the concomitant treatment with ganciclovir on the pharmacodynamics of LVFX-induced seizures to identify whether these factors can alter the pharmacokinetics or the pharmacodynamics of LVFX. Male Wistar rats received an intravenous infusion of LVFX at 250, 500, or 1000 mg/h/rat until the onset of seizures, and samples of serum, brain, and cerebrospinal fluid (CSF) were obtained. The concentration of LVFX in CSF at the onset of seizures was not affected by the infusion rate, whereas that in serum and brain increased with increasing infusion rate. This suggests that the concentration of LVFX in CSF is an appropriate index of the drug concentration at the site of action. The concentration of LVFX in CSF at the onset of seizures was significantly lower in rats with renal failure than in the control rats. Pretreatment with methylguanidine, an uremic toxin, at 600 mg/h/rat for 8 min reduced the concentration of LVFX in CSF at the onset of seizures and the total body clearance of LVFX after the intravenous injection. In rats pretreated with ganciclovir at 500 mg/h/rat for 1 h, the concentration of LVFX in CSF at the onset of seizures was significantly lower than the control rats. These results suggest that renal failure and ganciclovir can be the risk factors for LVFX induced seizures, and that they increase the sensitivity of the central nervous system to LVFX-induced seizures. PMID- 17409515 TI - Complex formation with plasmid DNA increases the cytotoxicity of cationic liposomes. AB - Cationic liposomes (CL) are one of the most widely studied non-viral vectors for gene delivery. It is well-known that CL induces cytotoxicity following lipofection. However, little is known regarding the mechanism involved in the cytotoxicity. In this study, the in vitro cytotoxicity of CL and its complex with pDNA (lipoplex) was investigated, and a part of the mechanism of induction as well. While free pDNA did not show any cytotoxicity, pDNA increased the cytotoxicity of CL via the formation of lipoplex. In addition, the lipoplex induced cytotoxicity increased in a lipoplex dose-dependent manner, irrespective of the type of pDNA, cell line and the absence or presence of serum. An assay showed that apoptosis was largely induced by treatment with the lipoplex (lipofection), but not with CL alone, in the tested range of concentration of CL and pDNA. Furthermore, following treatment with lipoplexes, the cells exhibited the morphological features of apoptosis and DNA fragmentation. A cDNA microarray study showed that the lipofection up-regulated 45 genes related to apoptosis, transcription regulation and immune response. These results clearly indicate that pDNA in the lipoplex increases the cytotoxicity of CL as a result of inducing apoptosis. The fundamental principle for gene therapy is to deliver gene-based therapeutics to target cells for specific gene targeting with minimal cytotoxicity. Our results suggest the possibility that cytotoxicity induced by lipofection, accompanied by gene changes, could intrinsically exacerbate, attenuate or even mask the desired effects of gene-based therapy. PMID- 17409516 TI - Efficient cytoplasmic protein delivery by means of a multifunctional envelope type nano device. AB - The potential for protein therapy, such as the use of antibodies, and vaccines is now well accepted. However, it is difficult to enhance efficiency in protein therapy without a suitable delivery system for delivering proteins to target sites. Here we describe the development of protein delivery system, which is capable of cytoplasmic delivery as well as efficient packaging. The multifunctional envelope-type nano device (MEND), which was originally developed for the delivery of nucleic acids such as plasmid DNA and oligodeoxynucleotides, can also be applied to protein delivery. In this study, the green fluorescent protein (GFP), a model protein, was condensed with stearyl octaarginine (stearyl R8) to form a nano particle, which was then coated with a lipid membrane, thus permitting R8 to be introduced for efficient cellular uptake and controlled intracellular trafficking. The packaging efficiency of the MEND was significantly higher than that of conventional liposomes, because the GFP can be encapsulated a condensed form. According to confocal laser scanning microscopy, the MEND is internalized efficiently and escapes from the acidic compartment to efficiently release GFP into the cytosol. These results indicate that the MEND can serve as a useful cytoplasmic delivery system for protein therapy. PMID- 17409517 TI - DNA microarray analysis of type 2 diabetes-related genes co-regulated between white blood cells and livers of diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats. AB - In a previous study, we hypothesized that some type 2 diabetes mellitus susceptible genes may be up/down-regulated in white blood cells (WBC) of Otsuka Long-Evans Tokushima Fatty (OLETF) rats, reflecting their up/down-regulation in major insulin-target tissues such as the liver before the onset of diabetes. We identified 57 potential candidate genes for predicting diabetes. In this study, we examined this hypothesis further by extending the experimental conditions from before the onset (6 weeks) to after the onset (24 weeks) of diabetes that type 2 diabetes mellitus susceptible genes are co-regulated in WBC, reflecting their expression in the liver. Using rat oligo DNA microarrays, we found that 48 genes are up/down-regulated in OLETF rats compared to control Long-Evans Tokushima Otsuka (LETO) rats in WBC and liver under fasting or insulin administration conditions. Twenty nine and 33 genes were up/down-regulated in both WBC and livers, respectively, under fasting and insulin administration conditions, respectively. Eight out of 29 genes in fasting condition and 12 out of 33 genes in insulin administration conditions have been reported to be type 2 diabetes mellitus susceptible genes and the remainder have not been reported to be related to type 2 diabetes mellitus. These results support our hypothesis that the expression levels of type 2 diabetes mellitus related genes in WBC are reflective of those in the liver after the onset of diabetes. PMID- 17409518 TI - Correlation between flavonoid content and the NO production inhibitory activity of peel extracts from various citrus fruits. AB - We investigated the correlation between the flavonoid content and NO production inhibitory activity of fruit peel extracts using 20 citrus plants. The contents of seven flavonoids (naringin, naringenin, hesperidin, hesperetin, rutin, nobiletin, and tangeretin) were determined by HPLC analysis. Each citrus peel extract varied in flavonoid content, but the contents of nobiletin and tangeretin, which were contained in all 20 fruit peels, showed a positive and significant correlation with each other (r=0.879, p<0.0005 for immature fruit peels; r=0.858, p<0.0005 for mature fruit peels). All citrus peel extracts dose dependently inhibited LPS-induced NO production in RAW 264.7 cells. This inhibitory effect was significantly and positively correlated with the content of nobiletin and tangeretin. Nobiletin showed a more potent NO production inhibitory activity (IC50=26.5 microM) compared to tangeretin (IC50=136.6 microM). This result supports the premise that nobiletin-rich citrus may provide protection against disease resulting from excessive NO production. PMID- 17409519 TI - Discrimination of Lonicera japonica THUNB. from different geographical origins using restriction fragment length polymorphism analysis. AB - Lonicera japonica THUNB. is a commonly used anti-inflammatory herbal medicine. The therapeutic effectiveness of L. japonica depends significantly on its geographical origin. However, it is difficult to define criteria for confirming geographical authenticity using microscopic and chemical characteristics. In the present study, the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA loci of L. japonica from different origins and related species was sequenced. The mutation site found in the ITS region from geo-authentic L. japonica can be recognized by the restriction endonuclease EcoN I. Since PCR products from geo-authentic L. japonica cannot be digested completely, a quantitative restriction fragment length polymorphism analysis method was developed. The cleavage rate of PCR products by EcoN I was determined to be more than 70% in all geo-authentic L. japonica and less than 20% in non-geo-authentic L. japonica and other species from genus Lonicera. The rate correlated remarkably with the geographical origin of L. japonica. Therefore, this method can be used to classify geo-authentic L. japonica. PMID- 17409520 TI - Induction of apoptosis by buckwheat trypsin inhibitor in chronic myeloid leukemia K562 cells. AB - Buckwheat is an ancient and specialty grain in China. Due to its unique chemical and bio-activity components, buckwheat has been found to have many uses in food products and medicine. However, very little is known about the toxicity of protease inhibitors from buckwheat. Here, the possible effects of a recombinant buckwheat trypsin inhibitor (rBTI) on the induction of apoptosis of the human K562 cell line were investigated by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide) assays and flow cytometric analysis. MTT assay showed that rBTI could specifically inhibit the growth of K562 cells in a dose dependent manner, but there were minimal effects on normal human peripheral blood mononuclear cells (PBMCs). Furthermore, comparison the effects of rBTI on K562 cells with those of negative control (BSA and the complex of BSA and rBTI) revealed that rBTI was highly toxic to K562 cells, and BSA hardly had any inhibition on proliferation in K562 cells. The analysis of flow cytometric indicated that the apoptosis of K562 cells were 31.0%, 32.8%, 35.3% and 52.1% after treated by rBTI in range of 12.5-100 microg/ml, respectively. The results suggested that rBTI can induce apoptosis of K562 cells and that it might be a potential protein drug of the trypsin inhibitor family. PMID- 17409521 TI - Enhancement effect of polyoxometalates on NGF-induced neurite-outgrowth of PC12 cells. AB - Nerve growth factor (NGF)-induced neurite-outgrowth of PC12 cells was enhanced by polyoxometalates such as Na9[SbW(9)O(33)].19.5H(2)O (SbW(9)) and (NH(3)Pr(i))6[Mo(7)O(24)].3H(2)O (Mo(7)). Western blotting analysis of polyoxometalate/NGF-treated PC12 cells showed a strong expression of growth associated protein 43 (GAP-43), which is associated with the neurite outgrowth. Similar effects were observed for other polyoxometalates, K(11)H[(VO)3(SbW(9)O(33))2].27H(2)O ((VO)3(SbW(9))2), K6[P(2)W(18)O(62)].14H(2)O (P(2)W(18)), and K7[PTi(2)W(10)O(40)].6H2O (PTi(2)W(10)), in contrast with little effect for monomeric tungstate and molybdate. Of the polyoxometalates tested in this study, SbW(9) and Mo(7) were the most potent. PMID- 17409522 TI - Induction of metallothionein synthesis by cilostazol in mice and in human cultured neuronal cell lines. AB - In this study, we examined the effect of Cilostazol to induce metallothionein (MT) in vivo and in vitro. Intraperitoneal injection of Cilostazol increased the expression of both MT-1 and MT-2 mRNA and total MT protein in the mouse liver. Cilostazol also augmented MT-1 mRNA levels in the murine brain. In vitro exposure to Cilostazol significantly augmented intracellular MT protein levels in cultured human brain microvascular endothelial cells (HBMEC) and in the neuroblastoma cell line IMR32. Taken together, these findings suggest that Cilostazol is an inducer of MT in the murine liver and brain, and that it has the potential to directly induce MT in cells. The contribution of the anti-oxidative effect of MT to the anti-stroke effect of Cilostazol was discussed. PMID- 17409523 TI - Differential antiviral activity of benzastatin C and its dechlorinated derivative from Streptomyces nitrosporeus. AB - Benzastatin C, a 3-chloro-tetrahydroquinolone alkaloid from Streptomyces nitrosporeus, showed antiviral activity in a dose-dependant manner with EC50 values of 1.92, 0.53, and 1.99 microg/ml against herpes simplex virus type 1 (HSV 1), herpes simplex virus type 2 (HSV-2), and vesicular stomatitis virus (VSV), respectively. In contrast, benzastatin D, the corresponding dechlorinated derivative, did not exhibit any antiviral activity. These results indicate that the antiviral activity of benzastatin C is mediated, in part, due to the chlorine moiety in its molecular structure. PMID- 17409524 TI - Molecular cloning and characterization of a multidrug efflux pump, SmfY, from Serratia marcescens. AB - We cloned a gene smfY for multidrug efflux pump from chromosomal DNA of Serratia marcescens using drug-hypersensitive Escherichia coli KAM32 as the host, and characterized the pump. E. coli KAM32/pESM42 carrying the smfY showed significantly increased MICs of various drugs including DAPI, norfloxacin, benzalkonium chloride, acriflavine and ethidium bromide, compared with the control. We also detected energy-dependent ethidium and acriflavine efflux due to the SmfY. Sequence analysis revealed that the SmfY was a multidrug efflux pump of the MF (Major Facilitator) superfamily transporters. This is the first report of a multidrug efflux pump belonging to the MF superfamily in S. marcescens. PMID- 17409525 TI - A microdialysis study of effects of gastrodin on neurochemical changes in the ischemic/reperfused rat cerebral hippocampus. AB - Gastrodin is a component extracted from the rhizome of Gastrodia elata, and has been shown to possess protective effects against neuron damage induced by simulated cerebral ischemia in previous studies. But its neurochemical effects on the ischemic brain had not been well studied. The present study aimed at evaluating the effects of gastrodin on the changes of transmitter amino acids in rat hippocampus during cerebral ischemia/reperfusion. Microdialysis sampling was performed during ischemia and early reperfusion periods in rats, and the glutamate and gamma-aminobutyric acid (GABA) in the dialysate were measured using high-performance liquid chromatography (HPLC). Administration of gastrodin (100 mg/kg) before ischemia significantly reduced the ischemia-induced elevation of glutamate levels during the postischemic period, increased the rise of extracellular GABA during the reperfusion periods, thus decreased the glutamate/GABA ratios during ischemia and reperfusion. These results provide insights to explain the neurochemical effects of gastrodin when applied prior to an ischemic event. PMID- 17409526 TI - Cudratricusxanthone A isolated from the root bark of Cudrania tricuspidata inhibits the proliferation of vascular smooth muscle cells through the suppression of PDGF-receptor beta tyrosine kinase. AB - Platelet derived growth factor (PDGF)-BB is one of the most potent vascular smooth muscle cell (VSMC) proliferative factors, and abnormal VSMC proliferation by PDGF-BB plays an important role in the development and progression of cardiovascular problems, including restenosis after coronary angioplasty and atherosclerosis. Previous phytochemical studies on the stems or root barks of Cudrania tricuspidata (Moraceae) resulted in the isolation of various isoprenylated xanthones and flavonoids, some of which have anti-cancer, hepatoprotective, anti-inflammatory and anti-oxidant activities. In the present study, we investigated the antiproliferative effect of cudratricusxanthone A isolated from the root bark of C. tricuspidata and its underlying mechanism in VSMCs. Antiproliferative effects of cudratricusxanthone A on VSMCs were examined by direct cell counting and [3H]-thymidine incorporation assays. Cudratricusxanthone A inhibited [3H]-thymidine incorporations into DNA in VSMCs that occurred in response to treatment with 50 ng/ml PDGF-BB. PDGF-BB-stimulated DNA synthesis was significantly reduced to 86.1, 80.2, 64.2 and 25.1% at concentrations of 0.1, 1, 2 and 3 microM, respectively. Moreover, pre-treatment with cudratricusxanthone A (0.1-3 microM) suppressed this PDGF-BB-stimulated cell number in a concentration-dependent manner. The inhibition percentages were 11.1, 22.7, 51.3 and 81.5% at the concentrations of 0.1, 1, 2 and 3 microM, respectively. We also investigated the mechanism of antiproliferative effects by cudratricusxanthone A in PDGF-BB-stimulated VSMCs. In Western blot analysis, 50 ng/ml PDGF-BB-stimulated phospholipase C (PLC)gamma1, Ras, and extracellular signal-regulated kinase1/2 (ERK1/2) phosphorylations were inhibited by cudratricusxanthone A (0.1-3 microM). Consisted with these findings, cudratricusxanthone A inhibited PDGF-receptor beta chain (PDGF-Rbeta) phosphorylation induced by PDGF-BB in a concentration-dependent manner. These findings suggest that the inhibitory effects of cudratricusxanthone A on DNA synthesis and proliferation by PDGF-BB-stimulated VSMCs are mediated by the suppressions of the PDGF-Rbeta and its downstream signaling pathways. Our observation may explain in part mechanistic basis for the prevention of cardiovascular diseases, such as atherosclerosis and restenosis after coronary angioplasty by cudratricusxanthone A. PMID- 17409527 TI - Diarylheptanoids from Alnus hirsuta inhibit the NF-kB activation and NO and TNF alpha production. AB - Six diarylheptanoids (1-6) from the stem bark of Alnus hirsuta were investigated for their inhibitory activity against LPS-induced NF-kB activation and NO and TNF alpha production. Among them, compounds 2, 3, and 6 displayed inhibitory activity against NF-kB activation and NO and TNF-alpha production with IC50 values of 9.2 9.9 microM, 18.2-19.3 microM, and 22.3-23.7 microM, respectively, in RAW264.7 cells. Three active compounds had no significant cytotoxicity in RAW264.7 cells at their effective concentrations. This is the first report of NF-kB-inhibitory activity of these compounds and supports the pharmacological use of A. hirsuta, which has been employed as a herbal medicine for the treatment of inflammatory diseases. PMID- 17409528 TI - Stimulatory activity of lignans from Machilus thunbergii on osteoblast differentiation. AB - Phytoestrogens are naturally occurring compounds exerting estrogenic activity, and include isoflavonoids, flavonoids and lignans. In the present study, we evaluated the stimulating activity of six lignans, meso-dihydroguaiaretic acid, nordihydroguaiaretic acid, machilin A, guaiacin, isoguaiacin and isoguaiacin dimethylether, from Machilus thunbergii, on osteoblast differentiation employing primary cultures of mouse osteoblast as an in vitro assay system. Among the six lignans tested, arylnaphthalene type lignans such as guaiacin, isoguaiacin and isoguaiacin dimethylether significantly increased alkaline phosphatase activity, whereas bibenzylbutane type lignans such as meso-dihydroguaiaretic acid, nordihydroguaiaretic acid and machilin A showed little effects. Isoguaiacin and isoguaiacin dimethylether also increased collagen synthesis as well as calcium deposition. In addition, treatment of the mouse osteoblasts with tamoxifen markedly reduced ALP activity increased by isoguaiacin or isoguaiacin dimethylether, suggesting the involvement of estrogen receptor in the action of these lignans on osteoblast differentiation. Taken together, these results suggest that arylnaphthalene type lignans such as guaiacin, isoguaiacin and isoguaiacin dimethylether significantly increase osteoblast differentiation. PMID- 17409529 TI - Sex differences in PPARgamma expressions in rat adipose tissues. AB - Clinical studies had suggested that there were sex differences in pharmacological and side effects of pioglitazone. However, there are few studies on the sex differences of peroxisome proliferator-activated receptor (PPAR) gamma expressions in rat adipose tissues. We investigated the sex differences in peroxisome proliferator-activated receptor (PPAR) gamma expressions in rat adipose tissues. Subcutaneous abdominal adipose tissues and perigonadal adipose tissues were obtained from male and female Wistar rats (12 weeks of age). Expressions of PPARgamma protein were determined by Western blot analysis with anti-PPARgamma antibody. Vaginal smear check was performed in female rats. We obtained adipose tissues from females, according to the different phases of the estrous cycle. Both PPARgamma1 and gamma2 subtypes were expressed in subcutaneous adipose tissues. Expressions of PPARgamma2 in subcutaneous adipose tissues were significantly higher in males than in females. On the other hand, expressions of PPARgamma2 in perigonadal adipose tissues were significantly higher in females than in males. Expressions of PPARgamma2 in perigonadal adipose tissues in females significantly decreased during diestrus. It can be suggested that the sex hormones might affect the expressions of PPARgamma2 in adipose tissues. PMID- 17409530 TI - Tissue-specific mRNA expression profiles of human carbohydrate sulfotransferase and tyrosylprotein sulfotransferase. AB - Pairs of forward and reverse primers and TaqMan probes specific to each of 15 human sulfotransferases were prepared. The mRNA expression level of each target enzyme was analyzed in total RNA from single and pooled specimens of various human tissues (adrenal gland, bone marrow, brain, colon, heart, kidney, liver, lung, pancreas, peripheral leukocytes, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thymus, thyroid gland, trachea, and uterus) by real-time reverse transcription PCR using an ABI PRISM 7700 Sequence Detection System. The mRNA expression profiles of the sulfotransferases in these 23 different human tissues were used to identify the tissues exhibiting high transcriptional activity for these enzymes. These results provide valuable information for studies concerning the human carbohydrate sulfotransferase and tyrosylprotein sulfotransferase genes in various tissues. PMID- 17409531 TI - Effect of ethanol on the binding of warfarin enantiomers to human serum albumin. AB - Ethanol is widely used as a pharmaceutical excipient for the solubilization of many hydrophobic drugs for injections. However, there are only few studies about drug interaction with pharmaceutical excipients in the body after injection. In this study, the effect of ethanol (500 mM) or several alcohols (500 mM) on the stereoselective binding of warfarin enantiomers to fatty acid-free human serum albumin (HSA) or proteins of commercial albumin preparations was investigated. An ultrafiltration method was used for the separation of unbound warfarin enantiomers. By the addition of ethanol or 1-propanol, the unbound fraction of the S-enantiomer was decreased. On the other hand, the unbound fraction of the R enantiomer was increased by the addition of ethanol or 1-propanol. Unbound fractions of both the S- and R-enantiomer were decreased by 2-propanol. In various commercial albumin preparations, unbound fractions of both the S- and R enantiomer were increased by ethanol. The different effects of ethanol among fatty acid-free HSA and commercial albumin preparations were observed. PMID- 17409532 TI - Percutaneous penetration kinetics of lidocaine and prilocaine in two local anesthetic formulations assessed by in vivo microdialysis in pigs. AB - The aim of this study was to characterize and compare the percutaneous penetration kinetics of lidocaine (L) and prilocaine (P) in two local anesthetic formulations by in vivo microdialysis coupled with HPLC. The microdialysis system for studying lidocaine and prilocaine was calibrated by a no-net-flux method in vitro and retrodialysis method in vivo, respectively. A dosage of 0.2 g/cm2 of an in-house P-L formulation (2.5% lidocaine and 2.5% prilocaine, methylcellulose based) and commercially available Eutectic Mixture of Local Anesthesia (EMLA, 2.5% lidocaine and 2.5% prilocaine, carbopol-based) was separately but symmetrically applied in the dorsal region of pigs. Saline (0.9%, w/v) was perfused into the linear microdialysis probe at a flow rate of 1.5 microl/min. Dialysate was collected upon topical application up to 6 h at 20-min intervals and assessed by HPLC. The results demonstrated the area under the concentration time curve (AUC(0-6 h)) of lidocaine and prilocaine in EMLA was 71.95+/-23.36 microg h/ml and 38.01+/-14.8 microg h/ml, respectively, in comparison to 167.11+/ 56.12 microg h/ml and 87.02+/-30.38 microg h/ml in the P-L formulation. The maximal concentrations (Cmax) of lidocaine and prilocaine in the dermis were 29.2+/-9.08 microg/ml and 16.54+/-5.31 microg/ml in EMLA and 80.93+/-17.98 microg/ml and 43.69+/-12.87 microg/ml in the P-L formulation, respectively. This study indicates a well-calibrated microdialysis system can provide vital real time information on percutaneous drug delivery and specifically a methylcellulose based P-L formulation can increase percutaneous absorption of both lidocaine and prilocaine in pigs compared to carbopol-based EMLA. PMID- 17409533 TI - Prevention of prion propagation by dehydrocholesterol reductase inhibitors in cultured cells and a therapeutic trial in mice. AB - In prion diseases, the normal cellular form of prion protein (PrP(C)) is converted into the disease-associated isoforms (PrP(Sc)) which accumulate in the infected tissues. Although the precise mechanism of this conversion remains unsolved, drugs of various categories have been reported to reduce the accumulation of PrP(Sc) in prion-infected cultured cells. We here show that AY 9944 (a 7-dehydrocholesterol reductase inhibitor) and U18666A (a 24 dehydrocholesterol reductase inhibitor) prevent PrP(Sc) from accumulating in prion-infected mouse neuroblastoma cells (ScN2a), with an ED50 of about 0.5 microM and 10 nM, respectively. In order to evaluate the efficacy of these two inhibitors in vivo, C57BL/6J mice inoculated with mouse-adapted scrapie-prion received repetitive intraperitoneal injections of U18666A (10 mg/kg) or a mixture of U18666A (10 mg/kg) and AY-9944 (12 mg/kg). By contrast to the potent anti prion effects observed in ScN2a cells, the in vivo trial was abortive with neither drug halting the progression of the disease. PMID- 17409534 TI - Genetic polymorphism of C452T (T127I) in human gamma-glutamyl hydrolase in a Japanese population. AB - We investigated the genotype distribution and allele frequency of C452T polymorphism of gamma-glutamyl hydrolase (GGH) gene, which causes the decreased enzymatic activity affecting the efficacy of methotrexate (MTX), in a Japanese population. The polymerase chain reaction-restriction fragment length polymorphism assay was applied to determine the genotype of C452T polymorphism in 269 Japanese healthy individuals. The genotype distribution was as follows: C/C, 89.2% (n=240); C/T, 10.4% (n=28); T/T, 0.4% (n=1). The frequency of C and T allele was 0.944 and 0.056, respectively. The obtained genotype distribution was well agreed with those expected by Hardy-Weinberg equilibrium. The genotype distribution and allele frequency in a Japanese population were found to be similar to those of African-Americans but significantly different from Caucasians. Although the frequency of variant T allele in a Japanese population is not so high as compared to Caucasians, determination of C452T polymorphism of GGH may be useful for monitoring of efficacy and side-effects of MTX for treatment of diseases such as rheumatoid arthritis or childhood acute leukemia. To our knowledge, this is the first report about the examination of C452T polymorphism of GGH in a Japanese population. PMID- 17409535 TI - Induction of various mutations during PCRs with manganese and 8-hydroxy-dGTP. AB - To induce various mutations randomly, PCRs with Mn2+ and with a mutagenic deoxyribonucleotide, 8-hydroxy-dGTP (8-OH-dGTP), were performed. Mutations were induced by deoxyribonucleotide imbalance plus 500 microM Mn2+ in the Mn2+-PCR, and the amplified DNA was inserted into a plasmid. The plasmid library obtained from the transformed bacterial cells was then used as the template in the next PCR, which was done with 50 or 100 microM 8-OH-dGTP. Four kinds of mutations, A:T ->G:C and G:C-->A:T transitions and A:T-->T:A and A:T-->C:G transversions, occurred with similar frequencies. These results suggest that this strategy will be useful in random PCR mutagenesis for the in vitro evolution of nucleic acids and proteins, and for analyses of residues in these biomolecules. PMID- 17409536 TI - Intake of fermented soybean (natto) increased locomotor activity in mice. AB - We fed mice food granules containing fermented soybean (natto in Japanese) powder (hereafter "natto granules") for 14 d to investigate whether natto granules had any effects on mouse behavior. We noted an enhancement of locomotor activity in natto-granule-fed mice compared to control and soybean-pellet-fed mice. This enhanced locomotor activity was blocked by a low dose of haloperidol (1 microg/kg i.p.), a dopamine receptor antagonist, but not by methysergide, a serotonin 5 HT(1/2) receptor antagonist. The results suggest that the enhanced locomotor activity induced by continuous intake of natto granules in mice is sensitive to haloperidol. PMID- 17409537 TI - Sensitive two-dimensional determination of small amounts of D-amino acids in mammals and the study on their functions. AB - D-amino acids are the candidates of novel physiologically active substances and the marker molecules of diseases in mammals. In the present study, the two dimensional determination of small amounts of D-amino acids in mammals has been performed after sensitive pre-column fluorescence derivatization with 4-fluoro-7 nitro-2,1,3-benzoxadiazole (NBD-F). The two-dimensional HPLC system includes the isolation of the target amino acids as D+L mixtures using the micro-ODS column, and the determination of the enantiomers using the chiral column. This method enables the sensitive and selective determination of small amounts of D-amino acids in mammals without the interference of L-amino acids and peptides, and the presence and distribution of D-Leu, D-Ala, D-Pro, D-Thr and D-allo-Thr has been demonstrated in rats and mice. The regulation and the origins of these D-amino acids, and their relationships to the biological rhythms are also discussed. PMID- 17409538 TI - Alpha-substituted 1-aryl-3-dimethylaminopropanone hydrochlorides: potent cytotoxins towards human WiDr colon cancer cells. AB - A series of 1-aryl-2-dimethylaminomethyl-2-propenone hydrochlorides 1 were prepared which possessed IC(50) values of less than 10 microM when examined towards human WiDr colon cancer cells. The related 1-aryl-2-dimethylaminomethyl-3 hydroxypropanone hydrochlorides 2, formed by hydration of the analogs in series 1, also had IC(50) values in the low micromolar range. On the other hand, conversion of 2-dimethylaminomethyl-1-(4-nitrophenyl)-2-propenone hydrochloride 1c into the corresponding 2-mercaptoethanol of adduct 3c led to a 37-fold reduction in potency. Two thirds of the compounds prepared in this study were more potent than a reference drug cisplatin while one third of these molecules displayed greater cytotoxicity to the WiDr cells than human CRL-2522 fibroblasts. A stability study of the 4-nitrophenyl analog in each of the series 1-3 in deuterium oxide was undertaken. In the case of 1c, replacement of the dimethylamino hydrochloride group by a hydroxy function was noted while in series 2, the loss of both water and dimethylamine hydrochloride gave rise to a mixture of two enones. The mercaptoethanol adduct 3c underwent deamination. The data obtained provide guidelines for amplifying the project in the future. PMID- 17409539 TI - Two new triterpenoids and a steroidal glycoside from the aerial parts of Ocimum basilicum. AB - Studies on the chemical constituents of the aerial parts of Ocimum basilicum have led to the isolation of three new compounds, basilol (1), ocimol (2), and basilimoside (3), along with two known constituents betulinic acid and oleanolic acid. The structures of the new constituents have been elucidated through spectral studies including 2D-NMR experiments (HMQC, HMBC, COSY, NOESY, and J resolved) and chemical transformation, as p-formylphenyl 3beta-hydroxyolean-12-en 28-oate (1), 2-methoxy-4-carbomethoxyphenyl 3beta-hydroxy-lup-20(29)-en-28-oate (2), and (22E)-24xi-ethyl-25-methylcholesta-5,22-diene-3beta-ol-3-O-D glucopyranoside (3). PMID- 17409540 TI - Dissociation of DNA from histone by reaction of anti-cancer drug cis diamminedichloroplatinum(II) with DNA-histone complexes used as cellular model. AB - Although both cis-diamminedichloroplatinum(II) (cisplatin or cis-DDP) and trans diamminedichloroplatinum(II) bind to DNA, only cis-DDP is widely used as a chemotherapeutic agent; the stereoisomer trans-DDP is inactive. DNA, generally, is wound around the histone core in the nucleus of living cells and forms the nucleosome structure. To understand the essentially different anticancer activities of cis-DDP and trans-DDP, it is necessary to investigate the interaction of cis-DDP (or trans-DDP) with DNA around the histone in the nucleosome. Here, we used psiX174DNA-histone(LNCaP) complexes prepared by the reaction of psiX174DNA with histone(LNCaP) extracted from LNCaP cells. We first show that the ability of cis-DDP to dissociate the DNA from psiX174DNA histone(LNCaP), as a nucleosome model, is much stronger than that of trans-DDP. As a result of the action of cis-DDP, the DNA in the nucleosome is rendered naked, and the naked DNA is vulnerable to cis-DDP (or other drugs). This study describes a new model showing that the difference in the activities of cis-DDP and trans-DDP is related to the difference in their abilities to dissociate the DNA from the nucleosome. PMID- 17409541 TI - Topological differences in the interaction of human DNA topoisomerase I with DNA histone complexes modified by cis- and trans-DDP. AB - We show that the trefoil, figure-eight knot, and mini circular closed DNA are formed by the reaction of cis-DDP-modified psiX174DNA-histone(LNCaP) complexes as a new nucleosome model with human DNA topoisomease I. The yields from cis-DDP modified complexes were far higher than that of trans-DDP. The topologically distinct invariant DNA such as the trefoil and figure-eight knot are not produced in the reaction of DNA topo I with psiX174DNA-histone(LNCaP) complexes that are not modified by platinum. Therefore, the anti-cancer activity of cis-DDP may be related to the production of the trefoil, figure-eight knot, and mini circular closed DNA forms in the living cell. We subsequently demonstrate that the yield mechanism and identification of the topologically-distinct invariant DNA can be explained by the topological method using a Jones polynomial and recombination through the topo I path intra-twisted looped DNA model. These results suggest that the distinguishing of anti-neoplastic activity of cis- and trans-DDP can be partially explained by the distinct topologies of DNA, trefoil, figure-eight knot, and mini circular closed DNA that they produce. PMID- 17409542 TI - The relationship between the chemical structures of dihydropyrazine derivatives and DNA strand-breakage activity. AB - Dihydropyrazine, a compound derived from sugars, possesses DNA strand-breakage activity. The relationship between the activity as assayed using pBR 322 ccc-DNA and the chemical structures of derivatives of dihydropyrazine (DHPs) has been investigated. The addition of Cu(2+) enhanced the activity remarkably. The introduction of a methyl or phenyl group onto the DHP ring or a cyclohexyl group fused onto the DHP ring also increased the activity. These properties indicated that the activity was due to the facility of electron release from the DHP ring, followed by radical generation. The determination of ionization potential and electrostatic potential values, and bond dissociation energy via semi-empirical MO calculations suggested strongly that the activity is induced by a DHP ring structure that contains a configuration suitable for hyperconjugation. PMID- 17409543 TI - Microbial transformation of 5-episinuleptolide. AB - 5-episinuleptolide (1) is an abundant norcembranolide diterpene isolated from several species of the soft coral genus Sinularia. Biocatalytic transformation studies of 1 using Streptomyces lavendulea ATCC 8664 resulted in the isolation and characterization of two new metabolites 2 and 3. Compound 2, 6alpha-hydroxy-5 episinuleptolide was produced via a stereoselective reduction of 1 and was further metabolized into compound 3 which has a 3,8-bicylized cembranoid skeleton. The structures and configurations of the metabolites were determined by spectroscopic and X-ray crystallographic analyses. PMID- 17409544 TI - Development and validation of spectrophotometric, TLC and HPLC methods for the determination of lamotrigine in presence of its impurity. AB - Three reliable, rapid and selective methods have been developed and validated for the determination of lamotrigine in the presence of its impurity, 2,3 dichlorobenzoic acid. The first method is spectrophotometric method using p chloranilic acid forming a colored product with lambda(max) 519+/-2 nm. All variables affecting the reaction have been investigated and the conditions were optimized. Beer's law was obeyed over a concentration range of 10-200 microg ml( 1) with mean accuracy 100.13+/-0.44%. The molar ratio of the formed ion association complex is found to be 1 : 1 as deduced by Job's method. The conditional stability constant (K(f)), standard free energy (DeltaG), molar absorptivity(epsilon), and sensitivity index were evaluated. The second method is based on TLC separation of the cited drug (Rf=0.75+/-0.01) from its impurity (Rf=0.23+/-0.01) followed by densitometric measurement of the intact drug spots at 275 nm. The separation was carried on silica gel plates using ethyl acetate : methanol : ammonia 35% (17 : 2 : 1 v/v/v) as a mobile phase. The linearity range was 0.5-10 microg/spot with mean accuracy 99.99+/-1.33%. The third method is accurate and sensitive stability-indicating HPLC method based on separation of lamotrigine from its impurity on a reversed phase C(18) column, using a mobile phase of acetonitrile : methanol : 0.01 M potassium orthophosphate (pH 6.7+/-0.1) (30 : 20 : 50 v/v/v) at ambient temperature 25+/-5 degrees C and UV detection at 275 nm in an overall analysis time of about 6 min., based on peak area. The injection repeatability, intraday and interday repeatability were calculated. The procedure provided a linear response over the concentration range 1-12 microg ml( 1) with mean accuracy of 99.50+/-1.30%. The proposed methods were successfully applied for the determination of lamotrigine in bulk powder, in dosage form and in presence of its impurity. The results obtained were analyzed by ANOVA to assess that no significant difference between each of the three methods and the reported one. The validation was performed according to USP guidelines. PMID- 17409545 TI - Improved stability of OPALMON tablets under humid conditions. III: Application of the rotary vacuum drying method to dry Opalmon tablets. AB - In stability studies on moisture-resistant Opalmon tablets in press-through packages (PTP), which were placed in aluminum bags, we found that the degradation rate of the dextran formulation is faster than that of the lactose formulation. The fast degradation of the dextran formulation is attributed to residual moisture in the package because drying the tablets before packaging suppressed the degradation and there is a good correlation between the stability of the drug and the water-activity of the tablets. Therefore, we developed a new drying method for the tablets, i.e. the rotary vacuum drying method, and investigated the effects of the operating conditions such as heating temperature, rotation speed, and vacuum degree on the drying time, and the appearance of the tablets. Using the rotary vacuum drying method, the tablets were dried over a short time (30 min) on a mass production scale so that the water activity was less than 0.03. Furthermore, the tablets suffered negligible damage such as breaking and chipping during the drying process. These results indicate that the rotary vacuum drying method is useful for drying tablets on mass production scales. PMID- 17409546 TI - Two new steroid glycosides and a new sesquiterpenoid glycoside from the underground parts of Trillium kamtschaticum. AB - Two new steroid glycosides, named trikamsterosides A and B, and a new sesquiterpenoid glycoside named trikamsesuquiside A were isolated from the underground parts of Trillium kamtschaticum PALL. along with 18 known compounds comprising 12 steroids, one sesquiterpenoid glycoside, one phenylpropanoid, one flavonoid glycoside, and three phenylpropanoid sucrose esters. Their chemical structures were determined on the basis of spectroscopic data and chemical evidence. Among them, one phenylpropanoid sucrose ester showed almost the same radical-scavenging effect on 1,1-diphenyl-2-picrylhydrazyl as that of alpha tocopherol. PMID- 17409547 TI - Synthesis and antibacterial activity of some novel triazolothienopyrimidines. AB - A novel series of some novel 5-substituted-1,2,4-triazolo[4,3-c]8,9,10 trihydrocyclopenta/8,9,10,11,12-pentahydrocyclohepta[b]thieno[3,2-e]pyrimidin-3 thiones has been synthesized. The intermediates 4-chloro-2-substituted-5,6,7 trihydrocyclopenta/5,6,7,8,9-pentahydrocyclohepta[b]thieno[2,3-d]pyrimidines were prepared by warming 2-substituted-5,6,7-trihydrocyclopenta/5,6,7,8,9 pentahydrocyclohepta[b]thieno[2,3-d]pyrimidin-4[3H]-ones with oxalyl chloride. Thieno[2,3-d]pyrimidin-4[3H]-ones were prepared by a novel, microwave assisted, solvent free, synthetic route under basic conditions hitherto unreported in the literature from ortho amino ester of thiophene. The chloro derivatives, without further purification, were hydrazinated to yield 2-substituted-4-hydrazino-5,6,7 trihydrocyclopenta/5,6,7,8,9-pentahydrocyclohepta[b]thieno[2,3-d]pyrimidines. These compounds were cyclized with carbon disulphide to give the title compounds in quantitative yields. The final compounds were screened for antibacterial activity by Kirby Bauer's method using ampicillin as the standard against various gram positive and gram negative bacteria. All the compounds showed antibacterial activity comparable with the standard. PMID- 17409548 TI - Studies on panax acetylenes: absolute structure of a new panax acetylene, and inhibitory effects of related acetylenes on the growth of L-1210 cells. AB - A new Panax acetylene, 3-oxo-PQ-1 (1), was isolated from Panax quinquefolium. The absolute configurations of 3-oxo-PQ-1 (1) and PQ-1 (2) were determined to be (9R,10R) and (3R,9R,10R), respectively, by synthesizing 1 and 2 starting from D-( )-diethyl tartrate, and by synthesizing their stereoisomers from L-(+)-diethyl tartrate. The growth inhibitory effects of Panax acetylenes (1-8) and their stereoisomers against leukemia cells were tested. Unnatural acetylenes having the (3S)-configuration (2, 5, 6, 7, 8; IC(50)=0.01-0.1 microg/ml) were found to be approximately ten times more potent than natural acetylenes (IC(50)=0.1-1.0 microg/ml) with the (3R)-configuration. Potency differences due to the configuration at C-9 and C-10 were unrelated to this stereochemistry. The C(14) polyacetylenes, PQ-8 (4) and its isomer (IC(50)=1.0-10.0 microg/ml), were found to exhibit weaker cytotoxicity than the C(17)-polyacetylenes. PMID- 17409549 TI - Inhibition of mannitol crystallization in frozen solutions by sodium phosphates and citrates. AB - Effects of co-solutes on the physical property of mannitol and sorbitol in frozen solutions and freeze-dried solids were studied as a model of controlling component crystallinity in pharmaceutical formulations. A frozen mannitol solution (500 mM) showed a eutectic crystallization exotherm at -22.8 degrees C, whereas sorbitol remained amorphous in the freeze-concentrated fraction in the thermal scan. Various inorganic salts reduced the eutectic mannitol crystallization peak. Trisodium and tripotassium phosphates or citrates prevented the mannitol crystallization at much lower concentrations than other salts. They also raised transition temperatures of the frozen mannitol and sorbitol solutions (T(g)': glass transition temperature of maximally freeze-concentrated amorphous phase). Crystallization of some salts (e.g., NaCl) induced crystallization of mannitol at above certain salt concentration ratios. Thermal and near-infrared analyses of cooled-melt amorphous sorbitol solids indicated increased intermolecular hydrogen-bonding in the presence of trisodium phosphate. The sodium phosphates and citrates should prevent crystallization of mannitol in frozen solutions and freeze-dried solids by the intense hydrogen-bonding and reduced molecular mobility in the amorphous phase. PMID- 17409550 TI - Medicinal flowers. XI. Structures of new dammarane-type triterpene diglycosides with hydroperoxide group from flower buds of Panax ginseng. AB - Six new dammarane-type triterpene diglycosides with a hydroperoxide group, floralginsenosides A, B, C, D, E, and F, were isolated from ginseng flower, the flower buds of Panax ginseng C. A. MEYER, together with seven known dammarane type triterpene oligoglycosides. The structures of new floralginsenosides were elucidated on the basis of chemical and physicochemical evidence. PMID- 17409551 TI - A novel benzofuran derivative, a new olean-type triterpene and other constituents from Ligularia odontomanes. AB - A novel benzofuran derivative (R)-(+)-Ligulaodonin A (1) and a new olean-12-ene triterpene (5), as well as 10 known compounds were isolated from the whole plant of Ligularia odontomanes HAND.-MAZZ. The structures of the new compounds were determined by spectroscopic evidence, including IR, UV, EI-MS, positive HR-ESI MS, 1D-NMR and 2D-NMR. The absolute configuration of 1 was elucidated on the basis of circular dichroism (CD) data. PMID- 17409552 TI - Design and gamma-scintigraphic evaluation of a floating and pulsatile drug delivery system based on an impermeable cylinder. AB - A blend of floating and pulsatile principles of drug delivery system seems to present the advantage that a drug can be released in the upper GI tract after a definite time period of no drug release. The objective of this study was to develop and evaluate a floating and pulsatile drug delivery system based on an impermeable cylinder. Pulsatile capsule was prepared by sealing the drug tablet and the buoyant material filler inside the impermeable capsule body with erodible plug. The drug delivery system showed typical floating and pulsatile release profile with a lag time followed by a rapid release phase. The lag time prior to the pulsatile drug release correlated well with the erosion properties of plugs and the composition of the plug could be controlled by the weight of the plug. The buoyancy of the whole system depended on the bulk density of the dosage form. Gamma-scintigraphic evaluation in humans was used to establish methodology capable of showing the subsequent in vivo performance of the floating and pulsatile release capsule. Developed formulations showed instantaneous floating with no drug release during the lag time followed by a pulse drug release. From the gamma-scintigraphic results, the pulsatile release capsule we prepared could achieve a rapid release after lag time in vivo, which was longer than that in vitro. The scintigraphic evaluation could confirm qualitatively that the system with in vitro lag time of 4.0 h provided, with relatively high reproducibility, a pulsatile release occurred around 5.0 h after administration. PMID- 17409553 TI - Bootstrap re-sampling technique to evaluate the optimal formulation of theophylline tablets predicted by non-linear response surface method incorporating multivariate spline interpolation. AB - Optimal solutions of theophylline tablet formulations were derived from three types of experimental datasets, composed of different numbers of data-points using the response surface method incorporating multivariate spline interpolation (RSM(S)). The reliability of these optimal solutions was evaluated by a bootstrap re-sampling technique. Different levels of three causal factors were used as factors of response surface analysis: the lactose/cornstarch ratio (X(1)), the amount of carmellose calcium (X(2)), and the amount of hydroxypropylcellulose (X(3)). The target responses were the dissolution ratio of theophylline for the first 15 min (Y(1)) and the hardness (Y(2)) of each of the prepared tablets. Similar optimal solutions were estimated in three different sizes of datasets. A bootstrap re-sampling with replacements from the original dataset was applied, and optimal solutions for each bootstrap dataset were estimated. The frequency of the distribution of the optimal solution generated by the bootstrap re-sampling technique demonstrated almost normal distribution. The average and standard deviation of the optimal solution distribution were calculated as evaluation indices reflecting the accuracy and reproducibility of the optimal solution. It was confirmed that the accuracy was sufficiently high, irrespective of the dataset size; however, the reproducibility worsened with a decrease in the number of the experimental datasets. Consequently, it was considered that the novel evaluation method based on the bootstrap re-sampling technique was suitable for evaluating the reliability of the optimal solution. PMID- 17409554 TI - Terpenoid-related metabolites from a formosan soft coral Nephthea chabrolii. AB - Two new sesquiterpenoidal natural products chabrolidiones A and B (1 and 2), two C(18) terpenoid-related carboxylic acids, ketochabrolic acid (3) and isoketochabrolic acid (4), and one naphthoquinone derivative chabrolonaphthoquinone C (5), along with two known compounds (+)-aristolone (6) and teuhetenone A (7) were isolated from a Formosan soft coral Nephthea chabrolii. The structures of the new metabolites were determined on the basis of extensive spectroscopic analysis and by comparison of NMR data with those of related metabolites. Metabolite 1 has been synthesized previously, but was isolated for the first time from natural sources. Cytotoxic activity of metabolites 1-3 and 5-7 against a limited panel of cancer cell lines is also described. PMID- 17409555 TI - Medicinal flowers. XIV. New acylated oleanane-type triterpene oligoglycosides with antiallergic activity from flower buds of chinese tea plant (Camellia sinensis). AB - The methanolic extract from the flower buds of Chinese tea plant (Camellia sinensis (L.) O. KUNTZE) was found to inhibit release of beta-hexosaminidase from RBL-2H3 cells. From the methanolic extract, six new acylated oleanane-type triterpene oligoglycosides, floratheasaponins D-I, were isolated together with 21 known compounds including floratheasaponins A-C. The chemical structures of floratheasaponins D-I were elucidated on the basis of chemical and physicochemical evidence. The principal constituents, floratheasaponins A-F, were found to show the inhibitory activity on the release of beta-hexosaminidase from RBL-2H3 cells. PMID- 17409556 TI - Medicinal flowers. XV. The structures of noroleanane- and oleanane-type triterpene oligoglycosides with gastroprotective and platelet aggregation activities from flower buds of Camellia japonica. AB - The methanolic extract from the flowers buds of Camellia japonica L. (Theaceae) were found to exhibit potent inhibitory activities on ethanol- or indomethacin induced gastric mucosal lesions in rats. Through bioassay-guided separation, 28 noroleanane-type triterpene oligoglycosides, camelliosides A, B, and C, and an oleanane-type triterpene oligoglycoside, camellioside D, were isolated from the methanolic extract together with five known compounds. The absolute stereostructures of camelliosides were determined on the basis of chemical and physicochemical evidence, which included the structure revision of the nortriterpene aglycons (camellenodiol and camelledionol). The principal oligoglycosides, camelliosides A and B, showed platelet aggregation activity in addition to the gastroprotective effects on ethanol- or indomethacin-induced gastric mucosal lesions in rats. PMID- 17409557 TI - Synthesis and structure activity relationship studies of benzothieno[3,2-b]furan derivatives as a novel class of IKKbeta inhibitors. AB - As a novel class of IKKbeta inhibitors, a series of tricyclic furan derivatives was designed and synthesized based on the structure of known thiophene IKKbeta inhibitors. Among the various fused furan derivatives synthesized, a benzothieno[3,2-b]furan derivative 13a displayed potent inhibitory activity towards IKKbeta in enzymatic and cellular assays. The potent inhibitory activity originates from an intramolecular non-bonded S...O interaction which was confirmed by the X-ray structure of JNK3 with 16k. The introduction of further substituents on the core structure led to the discovery of the 6-alkoxy derivatives, which possessed a comparable IKKbeta inhibitory activity to 13a and an improved metabolic stability. Among these, appropriately lipophilic compounds 16a, h, i, and 13g (log D>2) were found to possess good oral bioavailability. PMID- 17409558 TI - Poly(lactic-co-glycolic acid) microspheres for the controlled release of huperzine A: in vitro and in vivo studies and the application in the treatment of the impaired memory of mice. AB - Huperzine A loaded poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres were prepared by an oil/water (o/w) solvent evaporation technique. With a decrease of the ratio of o/w from 1 : 100 to 1 : 50, the encapsulation efficiency was reduced about 4%. Increasing the PVA concentration from 0.5 to 2% reduced the percentage encapsulation efficiency of huperzine A from 60.7 to 47.4% and the particle size of microspheres from 84.2 to 26.2 microm. The addition of stearic acid improved the encapsulation efficiency, but also accelerated the in vitro release of hupezine A from microspheres. After i.m. administration of huperzine A loaded microspheres in mice, huperzine A was sustained released from the PLGA microspheres up to 12 d with a low initial burst. Passive avoidance test of mice showed that the microspheres formulation offered an improved therapeutic efficiency in the treatment of the impaired memory of the mice superior to injection gastric (i.g.) administration of huperzine A suspension at the same dose, whose therapeutic efficiency was similar as that of a 50% reduced dose of the microspheres formulation. PMID- 17409559 TI - Spectrophotometric determination of dopaminergic drugs used for Parkinson's disease, cabergoline and ropinirole, in pharmaceutical preparations. AB - Simple and reproducible spectrophotometric methods have been developed for determination of dopaminergic drugs used for Parkinson's disease, cabergoline (CAB) and ropinirole hydrochloride (ROP), in pharmaceutical preparations. The methods are based on the reactions between the studied drug substances and ion pair agents [methyl orange (MO), bromocresol green (BCG) and bromophenol blue (BPB)] producing yellow colored ion-pair complexes in acidic buffers, after extracting in dichloromethane, which are spectrophotometrically determined at the appropriate wavelength of ion-pair complexes. Beer's law was obeyed within the concentration range from 1.0 to 35 microg ml(-1). The developed methods were applied successfully for the determination of these drugs in tablets. PMID- 17409560 TI - Three new monoterpenoids from the fruit of Genipa americana. AB - Three new monoterpenoids, called genipacetal, genipamide, and genipaol, were isolated from the fruit of Genipa americana L. (Rubiaceae), along with the four known iridoids genipin, gardendiol, deacetyl asperulosidic acid methyl ester, and shanzhiside. Their chemical structures were determined on the basis of spectroscopic data. PMID- 17409561 TI - Novel flavonoids of Thelypteris torresiana. AB - In our continuing research on cytotoxic components from the Formosan pteridophyte Thelypteris torresiana (GAUD.) ALSTONONE, two new compounds, a novel flavonoid, flavotorresin (1), and a flavonoid diglycoside, multiflorin C (2), along with five known compounds, were isolated. The structural elucidation was established on the basis of spectroscopic data analysis. The possible biosynthetic pathway of the flavonoids from this fern is summarized. PMID- 17409562 TI - Electrochemical determination of meloxicam in pharmaceutical preparation and biological fluids using oxidized glassy carbon electrodes. AB - The adsorptive and electrochemical behaviors of meloxicam (MLC) was investigated on a glassy carbon electrode that was electrochemically treated by anodic oxidation at +1.8 V, following potential cycling in the potential range from -0.8 to 1.0 V vs. Ag/AgCl reference electrode. The resulting electrode showed a good activity to improve the electrochemical response of the drug. MLC was accumulated at an electrochemically activated glassy carbon electrode (phosphate buffer pH 6) in a certain time and then determined by linear sweep voltammetry. The oxidative peak currents showed a linear function in the concentration ranges of 0.02 to 10 microM using a 240 s preconcentration time. The preconcentration medium-exchange approach was utilized for the selective determination of the drug in spiked urine and plasma samples with satisfactory results. The recovery values of the proposed method obtained 105% (RSD 2.5%) and 100% (RSD 1.8%) for urine and plasma samples, respectively. Also, the proposed method has been successfully used for determination of MLC in tablets. PMID- 17409563 TI - Two new quinoline and tris(4-hydroxycinnamoyl)spermine derivatives from Microdesmis keayana roots. AB - Purification of a Microdesmis keayana hydromethanolic root extract led to the isolation of two new natural compounds, xanthoquininamide (6-hydroxyquinoline-4 carboxamide) and tris(4-hydroxycinnamoyl)spermine (N(5)-(p-coumaroyl)-N(1),N(14) diferuloylspermine) which was named keayanine. Their structures were determined using spectroscopic analyses (ESI-MS, 1D- and 2D-NMR). PMID- 17409564 TI - Pancreatic lipase-inhibiting triterpenoid saponins from Gypsophila oldhamiana. AB - Three new triterpenoid saponins, gypsosaponins A-C (1-3), were isolated from the roots of Gypsophila oldhamiana (Caryophyllaceae). Their structures were established as 3-O-beta-D-galactopyranosyl-(1-->2)-[beta-D-xylopyranosyl-(1-->3)] beta-D-glucuronopyranosyl quillaic acid 28-O-alpha-L-arabinopyranosyl-(1-->2) alpha-L-arabinopyranosyl-(1-->3)-beta-D-xylopyranosyl-(1-->4)-alpha-L rhamnopyranosyl-(1-->2)-beta-D-fucopyranoside (1), 3-O-beta-D-galactopyranosyl-(1 ->2)-[beta-D-xylopyranosyl-(1-->3)]-methyl-beta-D-glucuronopyranosyl gypsogenin 28-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl-(1-->4)]-alpha-L rhamnopyranosyl-(1-->2)-beta-D-fucopyranoside (2), and 23-O-beta-D-glucopyranosyl gypsogenic acid 28-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-glucopyranosyl-(1- >6)]-beta-D-glucopyranoside (3), on the basis of various spectroscopic analyses and chemical degradations. The biological activities of 1-3 were examined inhibitory activity against pancreatic lipase, which showed inhibition of 58.2%, 99.2% and 50.3% at concentration of 1 mg/ml, respectively. PMID- 17409565 TI - Hydrolyzed metabolites of thalidomide: synthesis and TNF-alpha production inhibitory activity. AB - Putative hydrolyzed metabolites of thalidomide were prepared and characterized, and their inhibitory activity on tumor necrosis factor (TNF)-alpha production in the human monocytic leukemia cell line THP-1 was evaluated. Alpha-(2 Carboxybenzamido)glutarimide was a more potent TNF-alpha production inhibitor than thalidomide. PMID- 17409566 TI - Flavonol galactoside caffeiate ester and homoisoflavones from Caesalpinia millettii HOOK. et ARN. AB - Chemical examination of the stems of Caesalpinia millettii HOOK. et ARN. led to the isolation of new flavonol glycoside caffeiate ester (1) and homoisoflavone (2), along with four known homoisoflavones: eucomin (3), bonducellin (4), 8 methoxybonducellin (5) and intricatinol (6). The structures of 1 and 2 were established to be tamarixetin 3-O-(6''-O-E-caffeoyl)-beta-D-galactopyranoside (1) and (Z)-7-hydroxy-8-methoxy-3-(4-methoxybenzyl) chroman-4-one (2) on the basis of detailed analyses of physical, chemical, and spectral data. Compounds 3-6 were isolated from this plant for the first time. PMID- 17409567 TI - Microbial transformation of dextromethorphan by Cunninghamella blakesleeana AS 3.153. AB - The capability of Cunninghamella blakesleeana AS 3.153 to transform CYP2D6 probe drug dextromethorphan was investigated. Metabolites produced by strain AS 3.153 were detected by liquid chromatography-tandem mass spectrometry (LC-MS(n)) and the metabolite dextrorphan was identified by reference to confirm its structure. The yield of dextrorphan produced by C. blakesleeana AS 3.153 was over 90%. Quinidine, a CYP2D6 selective inhibitor, was applied to investigate its effect on biotransformation. The concentration of quinidine was 4-folds higher than that of dextromethorphan and the yield of dextrorphan was reduced by 84%, which proved there was drug metabolism enzyme similar to CYP2D6 in C. blakesleeana AS 3.153. It is concluded that C. blakesleeana AS 3.153 can be used as the suitable model strain in vitro to mimic human CYP2D6 metabolism. PMID- 17409568 TI - Alternative procedure for charged derivatization to enhance detection responses of steroids in electrospray ionization-MS. AB - A derivatization procedure has been examined to enhance the electrospray ionization (ESI)-MS detectabilities of steroids that charged derivatization is not suitable for. The derivatization procedure with 2-hydrazinopyridine or isonicotinoyl azide was very effective for the sensitive detection of di oxosteroids or di-hydroxysteroids, respectively, and the detection limits of the resulting derivatives were as low as about 2 fmol. The derivatives also provided intense characteristic product ions in the MS-MS, which are expected to be usable for the selected reaction monitoring mode. PMID- 17409569 TI - Hydroperoxysterols from the tunicate Eudistoma sp. AB - Two 27C hydroperoxysterols, 7beta-hydroperoxycholesterol (1) and its stereoisomer 7alpha-hydroperoxycholesterol (2), were isolated from the lipophilic extracts of a Formosan tunicate belonging to the genus Eudistoma. The structures of sterols 1 and 2 were elucidated on the basis of extensive spectral data analyses. Cytotoxicity of sterols 1 and 2 against a limited panel of cancer cell lines is also described. Sterol 1 show weak inhibitory effects on human neutrophil elastase release. PMID- 17409570 TI - QSPR prediction of aqueous solubility of drug-like organic compounds. AB - A quantitative structure property relationship (QSPR) study was performed to develop a model that relates the structures of 150 drug organic compounds to their aqueous solubility (log S(w)). Molecular descriptors derived solely from 3D structure were used to represent molecular structures. A subset of the calculated descriptors selected using stepwise regression that used in the QSPR model development. Multiple linear regression (MLR) is utilized to construct the linear QSPR model. The applied multiple linear regression is based on a variety of theoretical molecular descriptors selected by the stepwise variable subset selection procedure. Stepwise regression was employed to develop a regression equation based on 110 training compounds, and predictive ability was tested on 40 compounds reserved for that purpose. The final regression equation included three parameters that consisted of octanol/water partition coefficient (log P), molecular volume (MV) and hydrogen bond forming ability (HB), of the drug molecules, all of which could be related to solubility property. Application of the developed model to a testing set of 40 drug organic compounds demonstrates that the new model is reliable with good predictive accuracy and simple formulation. The use of descriptors calculated only from molecular structure eliminates the need for experimental determination of properties for use in the correlation and allows for the estimation of aqueous solubility for molecules not yet synthesized. The prediction results are in good agreement with the experimental values. The root mean square error of prediction (RMSEP) and square correlation coefficient (R(2)) of prediction of log S(w) were 0.0959 and 0.9954, respectively. PMID- 17409571 TI - Allergy-preventive flavonoids from Xanthorrhoea hastilis. AB - Allergy-preventive activity was demonstrated for an extract of resins from Xanthorrhoea hastilis R. BR. in a search for allergy-preventive substances from natural sources. By bioassay-directed fractionation of this plant extract, a new flavanone, 3',5'-dihydroxy-7,4'-dimethoxyflavanone (1), and two new chalcones, 3,5,2'-trihydroxy-4,4'-dimethoxychalcone (2) and 5,2'-dihydroxy-3,4,4' trimethoxychalcone (3), were isolated together with five known compounds, 5' hydroxy-7,3',4'-trimethoxyflavanone (4), 3'-hydroxy-7,4'-dimethoxyflavanone (5), liquiritigenin 7-methyl ether (6), 4,2'-dihydroxy-4'-methoxychalcone (7) and sakuranetin (8). The structures of 1, 2 and 3 were elucidated by spectroscopic methods. All of these compounds showed allergy-preventive effects. PMID- 17409572 TI - An 18-norspirostanol saponin with inhibitory action against COX-2 production from the underground part of Trillium tschonoskii. AB - A novel 18-norspirostanol saponin (1), along with Trillenoside A (2), was obtained from the underground parts of Trillium tschonoskii MAXIM., collected in Shennongjia Forest District, China. Based on the chemical and spectroscopic evidences, their structures were determined as shown in Fig. 1. 1 and 2 displayed marked inhibitory action towards COX-2 production in macrophagocytes of the mouse abdominal cavity stimulated by LPS at 10 microg/ml. PMID- 17409573 TI - Microbial transformation of the steroidal alkaloid dictyophlebine by Rhizopus stolonifer. AB - The microbial transformation of a steroidal alkaloid, dictyophlebine (1) with Rhizopus stolonifer (ATCC 10404) afforded three oxidized metabolites 2-4. Compound 2 was found to be a new product. These metabolites were structurally characterized on the basis of modern spectroscopic techniques. Their inhibitory activity towards acetyl- and butyrylcholinesterase has been evaluated and the new product 2 has been found to be more potent than the parent compound and other metabolites. PMID- 17409574 TI - Studies on heterobifunctional cross-linking reagents, 6-maleimidohexanoic acid active esters. AB - 6-maleimidohexanoic acid N-hydroxysuccinimide ester has been used widely for preparation of enzyme immunoconjugates as a unique heterobifunctional cross linking reagent. Its heterobifunctional reactivity is good, but its ester portion hydrolyzes easily in the presence of water. Several 6-maleimidohexanoic acid active esters (6-maleimidohexanoic acid 4-nitrophenyl ester, 6-maleimidohexanoic acid N-hydroxy-5-norbornene-endo-2,3-dicarboximide ester, and 6-maleimidohexanoic acid pentafluorophenyl ester) were prepared and their reactivity and stability in an aqueous media were tested. Of the synthetic esters, the pentafluorophenyl ester exhibited the highest reactivity and stability in aqueous media. PMID- 17409575 TI - Aldosterone as a cardiovascular risk hormone. AB - The pathophysiological role of aldosterone in the development of cardiovascular disease has long been considered to be due its potent volume expansion/hypertensive effect mainly via mineralocorticoid receptor (MR) expressed in renal tubular epithelial cells. However, recent accumulating lines of evidence from clinical and experimental studies have suggested that direct cardiovascular effect of aldosterone contributes to the development of cardiovascular injury via MRs in non-epithelial tissue. A series of recent clinical studies have revealed that patients with primary aldosteronism have higher incidence of cardiovascular and renal complications than those with essential hypertension, and that aldosterone antagonism has cardiovascular protective effect in patients with heart failure independent from blood pressure. Numerous experimental studies have shown that both inflammation and oxidative stress play an initial and key role in the development of aldosterone-induced cardiovascular injury via non-epithelial MR activation. In this review, we discuss recent research progress in aldosterone and MR effects, with special emphasis on the pathophysiological role of aldosterone in cardiovascular diseases and the possible molecular mechanism(s) of cardiovascular injury by non epithelial MR activation. PMID- 17409576 TI - New therapeutic target for metabolic syndrome: PPARdelta. PMID- 17409577 TI - Exocytic process analyzed with two-photon excitation imaging in endocrine pancreas. AB - To elucidate the spatiotemporal profiles of final secretory stage, we have established two-photon extracellular polar tracer (TEP) imaging, with which we can quantify all exocytic events in the plane of focus within the intact tissues. With such technique, we can estimate the precise diameters of vesicles independently of the spatial resolution of optical microscope, and measure the fusion pore dynamics at nanometer resolution. At insulin exocytosis in the pancreatic islets, it took two seconds for the fusion pore to dilate from 1.4 nm in diameter to 6 nm in diameter, and such unusual stability of the pore may be due to the crystallization of the intragranular contents. Opening of the pore was preceded by unrestricted lateral diffusion of lipids along the inner wall of the pores, supporting the idea that this structure was mainly composed of membrane lipids. TEP imaging has been also applied to other representative secretory glands, and has revealed hitherto unexpected diversity in spatial organizations of exocytosis and endocytosis, which are relevant for physiology and pathology of secretory tissues. In the pancreatic islet, compound exocytosis was characteristically inhibited (<5%), partly due to the rarity of SNAP25 redistribution into the exocytosed vesicle membrane. Such mechanisms necessitate transport of insulin granules to the cell surface for fusion, and possibly rendering exocytosis more sensitive to metabolic state. Two-photon imaging will be powerful tools to elucidate molecular and cellular mechanisms of exocytosis and related disease, and to develop new therapeutic agencies as well as diagnostic tools. PMID- 17409578 TI - Coronary and systemic stent embolization during percutaneous coronary interventions: a single center experience. AB - OBJECTIVE: In this study, we attempted to analyze the incidence and outcomes of systemic and coronary stent embolizations during percutaneous coronary interventions and have described the treatment and retrieval methods used. METHODS: We retrospectively studied 24,038 consecutive coronary angiography procedures carried out at The Baskent University Adana Hospital from 1998 to present to determine the total number of stent embolization events. RESULTS: Among them, 4,797 were consecutive coronary stent operations and embolization was encountered in 14 cases (0.29%; 95% CI = 0.14-0.44%, P < 0.0001). The mean age of the patients was 61 +/- 8 years and 78% were men. Stent embolization occurred more frequently in cases with significant proximal angulation. Calcified lesions were not noted in any of the cases. In 7 out of 14 cases, stent embolization occurred at an unknown location and the clinical course was uneventful thereafter. Treatment and retrieval methods of the other 7 cases included the following: 1. Emergency cardiac bypass surgery (3 cases, 43%) 2. Advancement of a low profile delivery balloon through the stent, inflating the balloon, and replacing the stent at the lesion site (3 cases, 43%) 3. Crushing the stent against the coronary wall using another stent (1 case, 14%) 4. 4-loop snare (1 case, failed) None of the cases had bleeding that required transfusion. The stent was not crushed or deployed in the coronary artery causing major cardiac complication in any case. CONCLUSION: Systemic and coronary embolizations of stent procedures are rare. Consequences of coronary stent embolization can lead to prompt cardiac bypass surgery if the retrieval or deployment methods fail. Stent deployment or crushing techniques may be attempted before retrieval in patients who do not suffer from coronary thrombosis and myocardial infarction due to stent embolization. PMID- 17409579 TI - Initial and long-term outcomes of sirolimus-eluting stents for calcified lesions compared with bare-metal stents. AB - The aim of this study was to compare the initial and long-term outcomes of sirolimus-eluting stents (SES) and bare-metal stents (BMS) in patients with calcified lesions without performing rotational atherectomy. The subjects were 79 consecutive lesions (38 in the SES group and 41 in the BMS group) which were confirmed to have superficially calcified lesions by intravascular ultrasound. In all lesions, the stent was implanted after predilatation with a balloon. The patient characteristics were not different between the 2 groups. All procedures were successfully performed in both groups. Vessel area was significantly smaller in the SES group than in the BMS group (11.01 +/- 3.88 mm(2) versus 13.08 +/- 3.49 mm(2), P < 0.005), as was the lumen area (5.41 +/- 2.31mm(2) versus 6.48 +/- 2.04 mm(2), P < 0.005). Minimum stent area was significantly smaller in the SES group than in the BMS group (5.61 +/- 1.54 mm(2) versus 6.69 +/- 1.74 mm(2), P < 0.01). In cases in whom angiographic follow-ups were performed, the late loss was significantly smaller in the SES group than in the BMS group (0.19 +/- 0.49 mm versus 0.76 +/- 0.48 mm, P < 0.001). The restenosis rate was significantly lower in the SES group than in the BMS group (8.8% versus 33.3%, P < 0.05) and the TLR rate tended to be lower in the SES group (7.9% versus 19.5%). Stent thrombosis was not observed in either group. The results suggest that SES are more effective than BMS and can be used safely when treating calcified lesions if predilatation with a balloon is possible. PMID- 17409580 TI - Reduced adiponectin level is associated with severity of coronary artery disease. AB - Adipocyte-derived adiponectin has an antiatherosclerotic effect that acts independently of its antidiabetic effect. Plasma adiponectin levels are generally low in subjects with coronary artery disease. In this study, the relationship between the plasma adiponectin level and the severity of coronary artery disease, as assessed using the Gensini score, an index for the severity of coronary artery stenosis, was investigated. The subjects of the study were 104 patients (72 men and 32 women; BMI, 23.5 +/- 3.3 kg/m(2); age, 63.6 +/- 10.1 years) admitted to Tokyo University Hospital for coronary angiography. Plasma adiponectin levels were inversely correlated with the insulin resistance index HOMA-IR (P = 0.0127). The plasma adiponectin level was significantly associated with the Gensini score (P = 0.0332). After adjustment for conventional risk factors for cardiovascular diseases, the plasma adiponectin level tended to be inversely correlated with the Gensini score (P = 0.087). The measurement of plasma adiponectin levels may be useful for predicting the severity of coronary artery stenosis. PMID- 17409581 TI - Clinical usefulness of electrophysiologic study (EPS)-guided risk stratification for life-threatening arrhythmia in patients with heart failure. AB - BACKGROUND: Ventricular tachyarrhythmia is one of the most important factors determining the prognosis of patients with heart failure and sudden death can be observed even during stable therapy controlling clinical heart failure. In this study, the usefulness of electrophysiologic study (EPS) for the prediction of a future arrhythmic event was evaluated in patients with heart failure. METHODS AND RESULTS: The patient population consisted of 474 patients with a history of clinical heart failure but without an episode of spontaneous sustained ventricular tachycardia or fibrillation (VT/VF). A Holter ECG was performed in all patients, and 177 of the 474 patients underwent EPS because of a recording of nonsustained VT (NSVT, > 5 beats). When sustained VT/VF was inducible in EPS, the patient was assigned to implantation of a defibrillation device. The patients were divided into 3 groups, ie, 1) no NSVT (n = 297), 2) NSVT + no inducible VT/VF (n = 134), and 3) NSVT + inducible VT/VF (n = 43), and were followed-up for > 12 months. All patients were followed-up under standard therapy for heart failure. There were no significant differences in basic clinical characteristics and therapies among the 3 groups. During the follow-up period of 32 +/- 18 months, 56/474 patients suffered a VT/VF episode, ie, 21/297 in no NSVT, 14/134 in NSVT + no inducible VT/VF, and 21/43 in NSVT + inducible VT/VF patients (P = 0.032). All patients were rescued from sudden death among patients with an implanted defibrillator, but 11 patients without a defibrillator died. CONCLUSION: In patients with heart failure, future arrhythmic events could be predicted by EPS and Holter ECG. EPS-guided risk stratification seems to be useful in managing patients with heart failure. PMID- 17409582 TI - Effect of radiofrequency catheter ablation on Doppler echocardiographic parameters in patients with Wolff-Parkinson-White syndrome. AB - The aim of this study was to compare the conventional Doppler echocardiographic parameters before and after accessory pathway ablation in patients with Wolff Parkinson-White (WPW) syndrome. Thirty patients (19 males, 11 females) aged 35.5 +/- 14.4 years were enrolled in the study. All patients underwent successful radiofrequency catheter ablation (RFCA). Echocardiograhic examination was performed before and after RFCA. Aortic and pulmonary flows, diastolic early (E) and late (A) transmitral filling velocities, their velocity time integrals (VTI), mitral diastolic filling time (DFT), deceleration time (DT), isovolumic relaxation time (IVRT), aortic ejection time, and aortic VTI were assessed before and after RFCA. We found that the pulmonary valve opened earlier than the aortic valve when the accessory pathway was located on the right ventricular side (P = 0.02). Otherwise, if the accessory pathway was located on the left ventricular side, the aortic valve opened earlier (P < 0.01). Intervals between the onsets of aortic and pulmonary flows were shortened after RFCA (P = 0.01). We also observed prolongation of DFT (P < 0.001), increases in A velocity (P < 0.05) and its VTI (P < 0.01), as well as a decrease in the E/A ratio (P < 0.01) and shortening of aortic ejection time (P = 0.01) with restoration of AV conduction. We conclude that Doppler echocardiographic examination can provide clues about accessory pathway location and RFCA causes some significant changes in Doppler echocardiographic time intervals. These changes confirm that cardiac synchrony is restored after RFCA. PMID- 17409583 TI - Long-term efficacy of combination therapy with losartan and low-dose hydrochlorothiazide in patients with uncontrolled hypertension. AB - We evaluated the long-term efficacy of losartan and low-dose hydrochlorothiazide combination therapy in the treatment of hypertension. We enrolled 15 Japanese hypertensive outpatients whose 24-hour ambulatory blood pressure was >or= 135/80 mmHg after candesartan 8 mg (CND group; n = 10) monotherapy or amlodipine 5 mg (AML group; n = 5) monotherapy for 2 months or more. The monotherapy was then switched to losartan 50 mg and hydrochlorothiazide 12.5 mg combination therapy. Ambulatory blood pressure and indices of glucose and lipid metabolism were measured at the end of the monotherapy and after 3 and 12 months of the combination therapy. In the CND group, 24-hour blood pressure decreased significantly from 137 +/- 9/89 +/- 4 to 126 +/- 8/81 +/- 7 mmHg after 3 months (P < 0.05/ P < 0.001) and to 123 +/- 7/81 +/- 4 mmHg after 12 months (P < 0.01/P < 0.001). In the AML group, 24-hour blood pressure decreased significantly from 137 +/- 11/81 +/- 7 to 125 +/- 12/75 +/- 6 mmHg after 3 months (P < 0.05/P < 0.05) and to 124 +/- 9/77 +/- 7 mmHg after 12 months (P < 0.05/NS). There were significant decreases in systolic blood pressure during the daytime (6:00-21:30), nighttime (22:00-5:30) and early morning (6:00-8:00) after 12 months in both groups. No adverse changes in the indices of glucose or lipid metabolism were observed in either group. In conclusion, long-term combination therapy with losartan and low-dose hydrochlorothiazide was effective in the treatment of hypertensive patients whose blood pressure was not controlled by candesartan or amlodipine monotherapy alone. PMID- 17409584 TI - Funduscopic examination has limited benefit for management of hypertension. AB - Hypertension (HT) increases the risks of major cardiovascular events and affects a majority of elderly populations. Thus, blood pressure control is the mainstay for prevention of cardiovascular diseases. However, there is only a limited number of parameters for management of HT. This study was performed on consecutive patients between the ages of 35 and 70 years with normotension (NT) and HT in order to determine the possible consequences of HT on retinal vasculature and to avoid debility-induced weight loss. We studied 120 patients (64 females, 54.3 +/- 9.3 years) with NT and 147 (81 females, 57.3 +/- 11.0 years) with HT. No case of grade III or IV hypertensive retinopathy (HR) was detected in the groups. Although the prevalences of grades I and II HR were higher and grade 0 HR was lower in the hypertensive group, respectively (P < 0.001 for all), differences according to obesity, diabetes mellitus, hypertriglyceridemia (P < 0.001 for all), and dyslipidemia (P < 0.01) were also significant between the 2 groups. Therefore, only 18.3% (27 cases) of the hypertensives, even in whom the effect of aging could not be excluded, had HT only in the absence of any other risk factor of atherosclerosis. Despite the relatively higher specificities of grades III and IV HR for HT, particularly for hypertensive crisis, funduscopic examination has limited benefit in the management of HT due to the very low prevalences of grades III and IV HR, the multifactorial backgrounds of grades I and II HR, and the association of the multiple risk factors of atherosclerosis in a majority of the hypertensive cases. PMID- 17409585 TI - Increased serum bilirubin levels coincident with heart failure decompensation indicate the need for intravenous inotropic agents. AB - We have reported that chronic heart failure (HF) patients with increased serum bilirubin coincident with acute decompensation have a poor prognosis, indicating severe congestion and low tissue perfusion. The aim of this study was to analyze the possibility of increased bilirubin coincident with acute decompensation as a parameter which indicates the need for intravenous inotropic agents. We stratified 131 decompensated chronic HF patients with a LVEF or= 1.2 mg/dL), intravenous inotropics contributed to significantly more abundant diuresis, body weight reduction, and decreases in bilirubin and serum creatinine in the first 5 in hospital days compared to those without (group A: inotropics +; n = 24 versus group B: -; n = 38: 1726 +/- 418 versus 1458 +/- 424 mL/day: P < 0.05, -3.1 +/- 1.6 versus -2.1 +/- 2.2 kg: P < 0.05, -0.74 +/- 0.51 versus -0.04 +/- 0.60 mg/dL: P < 0.01, -0.29 +/- 0.89 versus -0.01 +/- 0.24 mg/dL: P < 0.01), in spite of no significant difference in the doses of diuretics between the 2 groups. On the contrary, patients with low bilirubin (T-Bil < 1.2 mg/dL) recovered from decompensation equally irrespective of inotropic administration (group C: inotropics +; n = 15 versus group D: -; n = 54: 1557 +/- 329 versus 1507 +/- 406 mL/day, -2.9 +/- 1.7 versus -2.8 +/- 1.5 kg, -0.01 +/- 0.25 versus -0.08 +/- 0.23 mg/dL, 0.02 +/- 0.24 versus 0.47 +/- 0.19 mg/dL; NS, respectively). Inotropics were administered after all because of unimproved hemodynamics in 26% of group B patients, compared to 4% of group D patients (P < 0.01). Increased bilirubin coincident with HF decompensation can be a useful marker indicating the need for intravenous inotropic agent administration. PMID- 17409586 TI - Brief ischemia-reperfusion performed after prolonged ischemia (ischemic postconditioning) can terminate reperfusion arrhythmias with no reduction of cardiac function in rats. AB - OBJECTIVE: Recently, it has been reported that ischemic postconditioning, a brief episode of ischemia-reperfusion performed after prolonged ischemia, can reduce ischemic myocardial injury. However, the effects of ischemic postconditioning on ischemia/reperfusion injury remain unclear. We investigated the effects of brief ischemia-reperfusion before (ischemic preconditioning) and after (ischemic postconditioning) prolonged ischemia on myocardial ischemia/reperfusion injury, especially reperfusion arrhythmias. METHODS: Adult male Sprague-Dawley rats weighing about 400-500 g were used. The isolated heart was perfused using a working heart method (Krebs-Henseleit bicarbonate buffer). In the control group, after stabilization, diastolic global ischemia for 15 minutes was produced by a one-way ball valve with electrical pacing (330 bpm, 2.0 V). After ischemia, the heart was reperfused for 20 minutes. In the preconditioning and postconditioning groups, 5-minute global ischemia was produced before and after ischemia for 15 minutes with a 1 minute interval. An electrocardiogram was performed and left ventricular pressure (LVP, +dP/dt, -dP/dt) and CK activity in coronary effluent were measured during the protocol. RESULTS: Ischemic preconditioning did not affect the incidence or duration of reperfusion ventricular arrhythmias. Ischemic postconditioning could terminate reperfusion ventricular arrhythmias completely and reduced the duration of reperfusion ventricular arrhythmias significantly (P < 0.01). Furthermore, the recovery ratio of +dP/dt at 20 minutes after initial reperfusion was significantly higher in the postconditioning group than in the other groups. CONCLUSION: These results suggest that ischemic postconditioning can terminate reperfusion arrhythmias with no reduction of cardiac function, and may be useful for correcting stunned myocardium. PMID- 17409587 TI - Superior logistic model for decay of Ca2+ transient and isometric relaxation force curve in rabbit and mouse papillary muscles. AB - A decrease in myocardial intracellular calcium concentration ([Ca(2+)](i)) precedes relaxation, and a monoexponential function is typically used for fitting the decay of the Ca(2+) transient. However, a logistic function has been shown to be a better fit for the relaxation force curve, compared to the conventional monoexponential function. In the present study, we compared the logistic and monoexponential functions for fitting the [Ca(2+)](i) declines, which were measured using the aequorin method, and isometric relaxation force curves at 4 different onsets: the minimum time-derivative of [Ca(2+)](i) (d[Ca(2+)](i)/dt (min)) and force (dF/dt(min)), and the 10%, 20% and 30% lower [Ca(2+)](i) levels and forces over the data-sampling period in 7 isolated rabbit right ventricular and 15 isolated mouse left ventricular papillary muscles. Logistic functions were significantly superior for fitting the [Ca(2+)] (i) declines and relaxation force curves, compared to monoexponential functions. Changes in the normalized logistic [Ca(2+)] (i) decline and relaxation force time constants at the delayed onsets relative to their 100% values at d[Ca(2+)] (i)/dt(min) and dF/dt(min) were significantly smaller than the changes in the normalized monoexponential time constants. The ratio of the logistic relaxation force time constant relative to the logistic [Ca(2+)](i) decline time constant was significantly smaller in mouse than in rabbit. We conclude that the logistic function more reliably characterizes the [Ca(2+)](i) decline and relaxation force curve at any onset, irrespective of animal species. Simultaneous analyses using the logistic model for decay of the Ca(2+) transient and myocardial lusitropism might be a useful strategy for analysis of species-specific myocardial calcium handling. PMID- 17409588 TI - Impact of diabetes on cardiomyocyte apoptosis and connexin43 gap junction integrity: role of pharmacological modulation. AB - The integrity of myocardial structures plays a crucial role in signal transductions and cardiac function. The aim of this study was to test the hypothesis that diabetes mellitus (DM) exerts adverse effects on the integrity of gap junctions (GJs) and induces cellular apoptosis in rat cardiomyocytes that can be abolished by simvastatin or losartan therapy. An experimental model of DM (induced by streptozocin 60 mg/kg body weight) in adult male rats (n = 24) was utilized to investigate the integrity of GJs containing connexin43 (Cx43) and the incidence of cellular apoptosis in the left ventricular myocardium. These rats were divided into 3 groups; group I (insulin therapy only), group II (insulin plus simvastatin 20 mg/kg/day), and group III (insulin plus losartan 20 mg/kg/day). Diabetic rats and 8 healthy rats (group IV) were sacrificed at 3 weeks following DM induction for immunofluorescence analysis. The experimental results demonstrated that the number of intact Cx43 GJs and the integrated area (mum(2)) constituted by clusters of Cx43 spots were significantly higher in groups II and IV than in group III, and in groups II-IV than in group I (all P values < 0.05). Additionally, the number of apoptotic bodies was remarkably higher in group I than in groups II-IV, and notably higher in groups II-III than in group IV (all P values < 0.05). Simvastatin is more effective than losartan at inhibiting the effects of DM on the integrity of myocardial ultrastructures. Both drugs effectively prevent cellular apoptosis in diabetic rat heart. PMID- 17409589 TI - Pulmonary vein isolation for atrial fibrillation in patients with paroxysmal atrial fibrillation and prolonged sinus pause. AB - BACKGROUND: Symptomatic prolonged sinus pauses upon termination of atrial fibrillation (AF) are an indication for pacemaker implantation. METHODS AND RESULTS: We evaluated the clinical outcomes of 4 patients who showed prolonged sinus pauses (> 2 seconds) upon termination of AF and thus underwent ablation. The ablative procedure included pulmonary vein isolation, superior vena cava isolation, and cavo-tricuspid isthmus ablation. Twenty-four-hour ambulatory electro-cardiogram monitoring was performed before and 1 month after ablation. The maximum sinus pause decreased from 4.5 +/- 2.1 seconds before ablation to 1.7 +/- 0.2 seconds after ablation. Sinus pauses > 2.0 seconds disappeared after ablation in all 4 patients. Minimum heart rate increased from 35.0 +/- 8.1 beats/minute before ablation to 52 +/- 6.7 beats/minute after ablation. The number of heart beats in 24 hours did not change significantly after ablation. CONCLUSION: Prolonged sinus pauses after paroxysmal AF may result from depressed sinus node function, which can be eliminated by curative ablation of AF. PMID- 17409590 TI - Supraventricular tachycardia originating from the posterior atrioventricular node in the univentricular heart with single atrium. AB - Supraventricular tachycardia (SVT) was observed in a 13-year-old male patient with complex clinical features that included univentricular heart with single atrium, pulmonary atresia, and polysplenia syndrome. During electrophysiologic study, atrial burst stimuli reproducibly induced and terminated the SVT, while the occurrence of ventriculoatrial block did not interrupt the SVT. His bundle electrograms (HBEs) were recognized both in the anterior and posterior regions on the common atrioventricular (AV) valve annulus. The posterior His bundle activation was progressively delayed along with the shortening of atrial pacing cycle length until it finally lagged behind local ventricular activation. Thus, antegrade AV conduction was solely via the anterior AV node. In contrast, during the SVT, the earliest activation was observed in the posterior HBE. These observations suggested that the posterior AV node serves as an origin of the SVT and that two AV nodes were linked together possibly through a sling at the infra Hisian level. Radiofrequency catheter ablation applied to the posterior HBE eliminated the SVT. PMID- 17409591 TI - Long-term survival in double inlet left ventricle combined with pulmonary stenosis and parachute mitral valve: a rare case. AB - Univentricular heart is a rare and complex congenital disease. Its prognosis is usually poor without surgical intervention and long-term survival into adulthood is rare. We report a 41-year-old man with a double inlet left ventricle with pulmonary stenosis and a parachute mitral valve verified by echocardiography and magnetic resonance imaging (MRI). PMID- 17409592 TI - Massive pulmonary thromboembolism in pregnancy rescued using transcatheter thrombectomy. AB - We report a rescued 37-year-old woman in her 30(th) week of gestation with massive pulmonary thromboembolism who was admitted to our cardiac care unit with progressive dyspnea and 2 episodes of syncope. Helical chest CT showed massive pulmonary thromboembolism of both pulmonary arteries. Although 26,000 U/day of heparin was administered following insertion of a temporary filter, hemodynamic evaluation documented no improvement. Since pulmonary artery (PA) pressure increased from 62/22 mmHg to 80/24 mmHg just after an emergency cesarean section on day 2, an emergency transcatheter thrombectomy was performed and it showed decreased PA pressure following extensive thrombus aspiration. Mother and baby were discharged with no complications. PMID- 17409593 TI - Therapies for raising high-density lipoprotein cholesterol. PMID- 17409594 TI - The association between microalbuminuria and metabolic syndrome in the general population in Japan: the Takahata study. AB - OBJECTIVE: The metabolic syndrome is associated with an increased risk of chronic kidney disease, cardiovascular disease and mortality. However, the association between microalbuminuria and the metabolic syndrome has not yet been reported in the general population in Japan. Therefore, we undertook a population-based study to examine the association between microalbuminuria and the metabolic syndrome in Takahata, Japan. METHODS: Subjects of this cross-sectional study were individuals aged from 40 to 87 years old. The metabolic syndrome was defined according to the criteria of the Adult Treatment Panel III. Microalbuminuria was defined as a urine albumin-creatinine ratio of 30 to 300 mg/g. RESULTS: A total of 2,321 subjects (mean age 64 years old) were entered into the final analysis. Among them, the prevalence of the metabolic syndrome and microalbuminuria was 16.5% and 13.7%, respectively. There was a significantly positive correlation between the number of components of the metabolic syndrome and the corresponding prevalence of microalbuminuria (p<0.001). In the subjects with metabolic syndrome compared with those without metabolic syndrome, the age- and gender-adjusted odds ratio of microalbuminuria was 1.99 (95% CI, 1.49-2.66). Multiple logistic regression analysis revealed that high glucose, high blood pressure and obesity were independently associated with microalbuminuria. CONCLUSIONS: Our study revealed a strong relationship between microalbuminuria and the metabolic syndrome in the general population in Japan. More comprehensive and intensive management of the metabolic syndrome at its early stage is important to prevent the progression of renal injury and cardiovascular complications. PMID- 17409595 TI - Blockade of IL-2 receptor suppresses HTLV-I and IFN-gamma expression in patients with HTLV-I-associated myelopathy/tropical spastic paraparesis. AB - OBJECTIVE: Th1 activation based on a high HTLV-I proviral load is one of the characteristic immunological abnormalities in the peripheral blood lymphocytes of patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). To clarify the cause of this abnormality with the potential to be one of the therapeutic targets, we analyzed the involvement of interleukin-2 (IL 2)/IL-2 receptor (IL-2R) signaling in HTLV-I and interferon-gamma (IFN-gamma), which is a representative Th1 cytokine, expression in peripheral blood CD4(+) T cells from HAM/TSP patients. PATIENTS AND METHODS: Twelve patients with HAM/TSP were included in the study. After the peripheral blood CD4(+) T cells were treated in cultures under the presence of each anti-IL-2Ralpha, beta,and gamma blocking antiboby for 48 hours, both HTLV-I p19 antigen and IFN-gamma levels in the culture supernatants were measured using ELISA methods. To check the influence on cell proliferation under these culture conditions, the numbers of viable cells were simultaneously determined by MTS assay. RESULTS: Treatment with anti-IL-2Ralpha blocking antibody, but not anti-IL-2Rbeta or anti-IL-2Rgamma blocking antibody, suppressed HTLV-I p19 antigen expression levels. In addition, treatment with all types of anti-IL-2R blocking antibodies also suppressed IFN gamma expression levels. All of the types of anti-IL-2R blocking antibodies did not inhibit the proliferation. CONCLUSION: These results indicate that IL-2/IL-2R signaling is involved in HTLV-I and IFN-gamma expression on peripheral blood CD4(+) T cells from HAM/TSP patients, suggesting that the interruption of this signaling has therapeutic potential against HAM/TSP in patients with the focus on the down-regulation of Th1 activation based on a high HTLV-I proviral load in the peripheral blood. PMID- 17409596 TI - Intravenous ciprofloxacin versus erythromycin in the treatment of Legionella pneumonia. AB - BACKGROUND: Erythromycin (EM) and rifampicin (RFP) have mainly been used to treat patients with Legionella pneumonia. Since intravenous ciprofloxacin (CPFX) became available in Japan from 2000, many reports have been published detailing successful treatment of Legionella pneumonia with CPFX. In this study, we compared the evolution of patients with Legionella pneumonia treated with CPFX to those treated with EM. METHODS: The study included nine patients treated with CPFX and eighteen patients treated with EM. Diagnosis of these patients was made by culture, PCR, urinary antigen assay or a serological method. A comparison was made of the patients' characteristics, severity of pneumonia, efficacy of each agent and the clinical course. RESULTS: No significant differences were observed between the two groups, in regard to age, gender, underlying disease or severity of pneumonia. In addition, the period of time from onset of the disease until appropriate therapy did not differ significantly between the two groups. In the CPFX group, all of the patients were cured and in the EM group 16 out of the 18 patients were cured. Although there were no significant differences, the time to apyrexia, normalization of leukocytosis and a 50% decrease in C-reactive protein (CRP) occurred within a relatively shorter time frame in the CPFX group than in the EM group (3.5 versus 4 days, 4 versus 5.2 days, and 2.9 versus 10.3 days, respectively). And, the duration of antibiotic treatment in the CPFX group was significantly shorter than in the EM group. CONCLUSION: CPFX was as effective as erythromycin in the treatment of Legionella pneumonia. The effects of treatment may appear relatively earlier and the duration of treatment was significantly shorter in patients treated with CPFX therapy than with EM therapy. PMID- 17409597 TI - Effectiveness of subcutaneous growth hormone in HIV-1 patients with moderate to severe facial lipoatrophy. AB - OBJECTIVE: To evaluate effect of recombinant human growth hormone (rhGH) among HIV-infected adults with moderate to severe facial lipoatrophy as a side effect of long-term antiretroviral treatment. DESIGN: A prospective open-label study METHODS: Twenty-five HIV-1 patients with moderate to severe facial lipoatrophy who had been on antiretroviral treatment for more than 18 months were enrolled. rhGH (5 mg) was given every other day for 6 months. After treatment was completed, the participants were followed up for 6 months. Facial lipoatrophy was evaluated by computed tomography at months 0, 3, 6 and 12. RESULTS: Nearly all participants (24 of 25) completed the study. The sum of bilateral soft tissue thickness at the level of zygomatics at months 0, 3, 6, 12 were 7.23, 8.59, 8.35, 8.60 mm, respectively. There was significant improvement from baseline in month 3 (p=0.009) and month 12 (p=0.021). In the 6 months of follow-up, the soft tissue showed no significant decrease. Several side effects including diarrhea, arthralgia, myalgia, mastalgia and hand numbness were seen, which were self limited and transient. CONCLUSION: rhGH is effective and relatively safe for moderate to severe facial lipoatrophy. Its effect was sustained at least for 6 months after the cessation of rhGH. PMID- 17409598 TI - Impact of gender on in-hospital mortality of patients with acute myocardial infarction undergoing percutaneous coronary intervention: an evaluation of the TAMIS-II data. AB - OBJECT: It is a matter of concern that women have higher in-hospital mortality rates than men with percutaneous coronary intervention (PCI), however, it is not yet clear whether significant gender differences exist. We studied the influence of gender on the characteristics and in-hospital mortality among patients with acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI). METHODS: We used data from 15 acute care hospitals included in the sample from the Tokai Acute Myocardial Infarction Study II (TAMIS-II), a prospective study of all patients admitted to these hospitals from 2001 to 2003 with a diagnosis of AMI. We abstracted the baseline and procedural characteristics from detailed chart reviews which included not only physician notes but also nursing notes, and a questionnaire which included baseline characteristics, procedural course and in-hospital mortality. Multivariate analysis was performed, controlling for age and other variables which were found to be significantly different between men and women by chi-square test or unpaired t test. PATIENTS: A total of 566 women and 2,048 men were included in the present study. RESULTS: There were gender differences in age, comorbid conditions, smoking status, body mass index, activities of daily livings, heart failure on presentation, duration of stay, angiographic data, transfer to ICU/CCU, and thrombolytic drugs. In univariate analysis, women had a higher in-hospital mortality rate than did men; however, this gender difference disappeared after adjustment for age and other variables. CONCLUSION: Our study demonstrated that women with AMI who undergo PCI do not have a higher in-hospital mortality rate than men. PMID- 17409599 TI - Decompensated lamivudine-resistant hepatitis B virus-related cirrhosis treated successfully with adefovir dipivoxil allowing surgery for hepatocellular carcinoma. AB - We describe a 64-year-old man with decompensated hepatitis B virus (HBV)-related cirrhosis who became resistant to lamivudine. He was started on adefovir at 10 mg daily while continuing lamivudine therapy. Several months later, his liver function improved and subsequently his ascites disappeared. The serum HBV-DNA level became undetectable 11 months later. Twenty months after the start of additional treatment with adefovir, one hepatocellular carcinoma (HCC) was detected, and the patient underwent a successful hepatectomy. Our findings suggest that the addition of adefovir to ongoing lamivudine therapy is useful for improving liver function in patients with decompensated lamivudine-resistant HBV related cirrhosis, allowing surgery for HCC. PMID- 17409600 TI - Primary mesenteric liposarcoma successfully diagnosed by preoperative imaging studies. AB - Liposarcoma is one of the most common primary neoplasms in the retroperitoneum, whereas primary mesenteric liposarcomas are rare. We encountered a case of liposarcoma which arose from the mesentery of the jejunum that was diagnosed by imaging studies before surgical operation. PMID- 17409601 TI - Autoimmune pancreatitis with extreme elevation of DUPAN-2. AB - An 80-year-old woman was admitted to our hospital with complaints of jaundice and liver dysfunction. She was found to have a high titer of serum IgG4, positive rheumatoid factor and marked elevation of DUPAN-2 (11,148 U/ml). Computed tomography showed swelling of the pancreas, and endoscopic retrograde cholangiopancreatography revealed diffuse irregular narrowing of the main pancreatic duct, which are typical findings of autoimmune pancreatitis. There was no evidence of malignancy. Administration of 30 mg/day of prednisolone was started. Computed tomography showed significant regression in the size of the pancreas, and the stenosis of the main pancreatic duct was improved on ERCP. The serum level of DUPAN-2 was also markedly decreased after the treatment. PMID- 17409602 TI - Serum adiponectin and leptin in a patient with Cushing's syndrome before and after adrenalectomy. AB - We measured the serum adiponectin and leptin concentrations before and after successful removal of a left adrenal adenoma in a 46-year-old woman with Cushing's syndrome. The serum adiponectin level was 6.0 microg/ml before the operation and rose to 8.1 microg/ml after adrenalectomy. However, the serum leptin level was markedly high (24.8 ng/ml) before the operation and decreased to within the normal range (6.0 ng/ml) 6 months after adrenalectomy, concomitant with weight reduction and normalization of the serum cortisol level. PMID- 17409603 TI - An elderly case of type 2 diabetes which developed in association with oral and esophageal candidiasis. AB - A 75-year-old woman who had been healthy except for mild glycemia and lipidemia discovered three and a half months before admission experienced severe dysphagia secondary to oral and esophageal candidiasis. She eventually developed diabetic hyperosmolar syndrome and ketoacidosis. Since anti-GAD antibody was negative and her diabetes was controlled with a moderate dose of insulin, we made a diagnosis of type 2 diabetes. Her only risk factors for candidiasis were hyperglycemia, age, and continuous denture use. The fact that her diabetes developed in association with oral candidiasis supports the hypothesis that there is a bidirectional interrelationship between diabetes and oral infection. PMID- 17409604 TI - Povidone iodine-induced overt hypothyroidism in a patient with prolonged habitual gargling: urinary excretion of iodine after gargling in normal subjects. AB - Iodine-induced hypothyroidism that develops in patients who gargle routinely with povidone iodine is well known. Usually the hypothyroidism is mild and resolves spontaneously upon cessation of gargling. Here, we report a 63-year-old patient with overt hypothyroidism that developed due to habitual gargling with povidone iodine for more than 10 years. The urinary excretion of iodine was estimated to be greater than 5 mg/day, based on values obtained from 18 normal subjects who gargled three times a day (4.6+/-2.1 mg, mean+/-SD). After discontinuation of the gargling, the patient has been euthyroid for more than 10 months. PMID- 17409605 TI - Acquired ichthyosis disclosing non-Hodgkin's malignant lymphoma. AB - When ichthyosis occurs in adulthood, it may signify internal disease, especially malignant hemopathy. A 49-year-old man was admitted to hospital with diarrhea, fever, weight loss and abdominal pain. Physical examination showed generalized ichthyosis with palmoplantar hyperkeratosis and multiple inguinal lymph nodes. Laboratory investigations showed severe pancytopenia. Ultrasonography and CT scan demonstrated intraabdominal lymph node enlargement. Osteomedullary biopsy and open abdominal lymph node biopsy revealed an anaplastic large cell lymphoma. The lymph node involvement subsequently became worse and more severe, and the patient's general condition deteriorated. He died before receiving chemotherapy. PMID- 17409606 TI - Fatal fulminant Clostridium difficile colitis during CHOP therapy for lymphoma: an autopsy case. AB - Although Clostridium difficile colitis is a common problem during chemotherapy, fulminant expression is rarely observed. Here, we describe a 68-year-old woman who developed fatal colitis due to Clostridium difficile infection. The patient was treated with CHOP therapy for relapsed lymphoma. In the nadir phase, she developed severe bloody diarrhea with a high fever and died within 12 hours after the beginning of symptoms. Clostridium difficile was identified in her stool and an autopsy showed hemorrhagic necrosis on the whole colon and rectum. This case demonstrates the substantial incidence of an unexpected feature with Clostridium difficile infection even with popular chemotherapy. PMID- 17409607 TI - Insulinoma with early-morning abnormal behavior. AB - We report a 65-year-old man with insulinoma who initially developed stereotypical behaviors and then progressed to more complex behaviors occurring early in the morning. He could not remember the events during the episodes. Insulinoma was diagnosed based on fasting blood glucose level of 15 mg/dl, high fasting immunoreactive insulin/blood glucose ratio (more than 0.3), and a tumor in the pancreas head by abdominal CT. Hypoglycemia caused by insulinoma should be considered as one of the mechanisms underlying abnormal nocturnal behaviors since the symptoms are very similar to those of sleep-related epilepsy, parasomnia, and night delirium. PMID- 17409608 TI - Two cases of refractory Wegener's granulomatosis successfully treated with rituximab. AB - Conventional therapy for Wegener's granulomatosis, steroid and cyclophosphamide, fails to control disease activity in some refractory patients and has treatment related toxicity. B cell depletion therapy using rituximab, a chimeric anti-CD20 monoclonal antibody, has been shown to be effective for certain autoimmune diseases including antineutrophil cytoplasmic antibody (ANCA) -associated systemic vasculitis. We report two refractory cases of Wegener's granulomatosis: one with bronchial and pulmonary involvement and retroorbital granuloma, the other with retroorbital granuloma and hypertrophic pachymeningitis causing severe headache. Rituximab was effective in both cases, with diminished granuloma and reduced ANCA titers, allowing steroids to be tapered. No adverse effects were detected. PMID- 17409609 TI - Tuberculous and syphilitic meningitis in a patient infected with the human immunodeficiency virus. AB - This report examines simultaneous tuberculous and syphilitic meningitis in a patient with human immunodeficiency virus (HIV). The 41-year-old homosexual patient presented with meningitis and hydrocephalus. His CD4+ and CD8+ T lymphocyte counts per microliter were 60 and 71, respectively. His clinical presentation was of a rapid-onset cerebral attack incompatible with tuberculous meningitis. Imaging indicated fatal bilateral cerebral infarction at the basal nuclei, and basal meningitis. We believe that syphilitic meningitis in HIV exacerbated the central nervous system (CNS) damage caused by tuberculosis and syphilis. The combination of three pathogens, Mycobacterium tuberculosis, Treponema pallidum, and HIV, has a great potential to cause serious CNS damage. PMID- 17409610 TI - Angioimmunoblastic T-cell lymphoma initially presenting with replacement of bone marrow and peripheral plasmacytosis. AB - A 73-year-old man presented with lymphadenopathy, hepatosplenomegaly, and a variety of hematological and immunological abnormalities. The bone marrow was replaced by polymorphic cellular infiltrates containing aggregates of CD10(+) T cells. Circulating lymphoplasmacytic/immunoblastic cells showed an early plasma cell immunophenotype on flow cytometric analysis. Combination of these observations indicated that the underlying disorder of this patient was angioimmunoblastic T-cell lymphoma (AITL); postmortem pathology was consistent with progression of peripheral T-cell lymphoma. Even in the absence of definitive lymph node biopsy, the appearance of the bone marrow and the peripheral blood can lead to the diagnosis of AITL. PMID- 17409611 TI - Lung fibrosis in carcinoid syndrome. PMID- 17409612 TI - Renal pelvic cancer revealed by renal biopsy. PMID- 17409613 TI - Bilateral endobronchial involvement in mantle cell lymphoma. PMID- 17409614 TI - [Esophagogram: diagnosis by a radiology technologist]. PMID- 17409615 TI - [Patent licensing (3)]. PMID- 17409616 TI - [Relationship between the criteria for metabolic syndrome and the evaluation of abdominal fat distribution measured by CT scan]. PMID- 17409617 TI - [Activities to promote medical safety (2006)]. PMID- 17409618 TI - [2006 standardization committee activities]. PMID- 17409619 TI - [Security measures for medical data under the personal information protection law]. PMID- 17409620 TI - [International activity of JIRA international division: administration organization and activity contents of international division?]. PMID- 17409621 TI - [Time-dependence evaluation of the primary visual cortex by 1H MR-spectroscopy using a stimulation control system]. AB - Proton magnetic resonance spectroscopy ((1)H MRS) is used to evaluate tumors and their remaining function under the assumption that event-related inspection and a method of data collection dependent on time are difficult. Therefore, this study examined the possibility that time depended on in MRS with a visual cortex. We conducted experiments using the following stimulations: REST: eyes are in a closed state, CONTROL: eyes are in an open state, and Continuous: consecutive stimulation and time dependence in the visual cortex. We enabled dependence in time by collecting data with a system that controlled stimulation and experimented on stimulation for a short time using this system. The results showed that no significant difference in metabolites was seen in the REST, CONTROL, and Consecutive stimulations. However, a significant difference was seen in Cr and NAA with time-dependent stimulation. Therefore, we considered functional magnetic resonance spectroscopy (fMRS) useful because time-dependent stimulation showed a meaningful difference with REST. PMID- 17409622 TI - [New method of determining regional fat fraction with modulus and real multiple gradient-echo (MRM-GRE)]. AB - We devised a method of analyzing a regional fat fraction using a modulus and real multiple gradient-echo (MRM-GRE) 4-7 echo Dixon sequence. The regional fat fraction was calculated with a theoretically fitted signal-intensity curve for multiple GRE images at each echo time (i.e., 4-7 echoes.) on a 1.5 Tesla MRI to correct T2* decay and phase cycling. Then the real image of the first echo was used to differentiate between the areas above and below the 50 percent fat fraction. To eliminate the T1-effect, the flip angle was set at 12 degrees. Fat fractions of the phantom with MRM-GRE were compared with those using conventional double-GRE and other MRI techniques. Fat fractions of liver and spinal bone marrow in healthy volunteers were determined during a breath-hold. The fat fraction of the phantom obtained by the MRM-GRE method tended to agree with the actual fat fraction. On the other hand, in both the phantom and in healthy volunteers, the double-GRE was underestimated owing to T2* decay. The MRM-GRE method enables simple, accurate analysis of the regional fat fraction. PMID- 17409623 TI - [Evaluation of the resolving power of different angles in MPR images of 16DAS MDCT]. AB - In this study, we evaluated the resolving power of three-dimensional (3D) multiplanar reformation (MPR) images with various angles by using 16 data acquisition system multi detector row computed tomography (16DAS-MDCT) . We reconstructed the MPR images using data with a 0.75 mm slice thickness of the axial image in this examination. To evaluate resolving power, we used an original new phantom (RC phantom) that can be positioned at any slice angle in MPR images. We measured the modulation transfer function (MTF) by using the methods of measuring pre-sampling MTF, and used Fourier transform of image data of the square wave chart. The scan condition and image reconstruction condition that were adopted in this study correspond to the condition that we use for three dimensional computed tomographic angiography (3D-CTA) examination of the head in our hospital. The MTF of MPR images showed minimum values at slice angles in parallel with the axial slice, and showed maximum values at the sagittal slice and coronal slice angles that are parallel to the Z-axis. With an oblique MPR image, MTF did not change with angle changes in the oblique sagittal slice plane, but in the oblique coronal slice plane, MTF increased as the tilt angle increased from the axial plane to the Z plane. As a result, we could evaluate the resolving power of a head 3D image by measuring the MTF of the axial image and sagittal image or the coronal image. PMID- 17409624 TI - [influence and improvement of metal artifacts in dental structures by CT for radiation treatment planning: reconstruction of transverse images using oblique images by gantry tilt scanning]. AB - Intensity-modulated radiation therapy (IMRT) radiation treatment planning (RTP) requires accuracy. Metal artifacts are one of the factors that influence RTP. The metal artifacts from dental structures are problems at the level of the oropharynx, since these artifacts impair visualization of tumors or lymph nodes and change CT (computed tomography) values. We simulated RTP at the level of the oropharynx using CT images with and without artifacts from dental structures. Gantry tilt scanning was performed to avoid artifacts from dental structures and transverse images reconstructed from oblique images by gantry tilt scanning using a technique of multiplanar reconstruction (MPR) . The reconstructed transverse images were used for the RTP. The reconstructed transverse images were useful to obtain accurate target volumes and the RTP of two opposed equally weighted beams by correct CT values. As dose distribution was changed slightly by the metal artifacts, the use of CT images without artifact is recommended in RTP. PMID- 17409625 TI - [Investigation of the present management status of calibration source based on the law concerning prevention of radiation hazards due to radioisotopes]. AB - An amendment concerning the enforcement of the law on the prevention of radiation hazards due to radioisotopes, etc., and the medical service law enforcement regulations were promulgated on June 1, 2005. This amendment concerned international basic safety standards and the sealing of radiation sources. Sealed radiation sources < or =3.7 MBq, which had been excluded from regulation, were newly included as an object of regulation. Investigation of the SPECT system instituted in hospitals indicated that almost all institutions adhere to the new amendment, and the calibration source, the checking source, etc., corresponding to this amendment were maintained appropriately. Any institutions planning to return sealed radioisotopes should refer to this report. PMID- 17409626 TI - [Simulation study of radiographic image quality measurement based on transmitted information]. AB - Although radiographic image quality is considered difficult to evaluate in a straightforward and systematic manner, it may be possible by using an index of transmitted information. As a preliminary study, relations between transmitted information and two image characteristics, namely, image noise and image blurring, were evaluated by simulation. The value of transmitted information was decreased if image noise and image blurring increased. The relationships were corroborated on an experimental basis. This paper suggests the possibility of a simple, straightforward method for synthetically evaluating radiographic images by a single parameter in terms of transmitted information. PMID- 17409627 TI - Edaravone, a radical scavenger, inhibits mitochondrial permeability transition pore in rat brain. AB - Edaravone, a radical scavenger, prevents ischemia/reperfusion injury in the brain, but the detailed mechanism is not known. This study examines the effect of edaravone on mitochondrial permeability transition pore (PTP) in rat brain. Edaravone at 10 - 100 microM inhibited Ca(2+)- and H(2)O(2)-induced swelling of mitochondria isolated from rat brain. Addition of Ca(2+) generated reactive oxygen species (ROS) in isolated mitochondria. Edaravone (10 - 100 microM) inhibited Ca(2+)-induced generation of ROS. These results suggest that edaravone inhibits opening of mitochondrial PTP in the brain, and they imply that inhibition of mitochondrial PTP may account for the neuroprotective effect of edaravone. PMID- 17409628 TI - Activation of mitogen-activated protein kinase by hepatocyte growth factor is stimulated by both alpha1- and beta2-adrenergic agonists in primary cultures of adult rat hepatocytes. AB - We investigated the effects of alpha(1)- and beta(2)-adrenergic agonists on hepatocyte growth factor (HGF)-stimulated mitogen-activated protein kinase (MAPK) isoforms in primary cultures of adult rat hepatocytes. Hepatocytes were isolated and cultured with HGF (5 ng/ml) and/or alpha- and beta-adrenergic agonists. Phosphorylated MAPK isoforms (p42 and p44 MAPK) were detected by Western blotting analysis using anti-phospho-MAPK antibody. The results show that HGF increased phosphorylation of p42 MAPK by 2.2-fold within 3 min. The HGF-induced MAPK activation was abolished by AG1478 treatment (10(-7) M). The MEK (MAPK kinase) inhibitor PD98059 (10(-6) M) completely inhibited the HGF-dependent increase in MAPK activity. Phenylephrine (10(-6) M) and metaproterenol (10(-6) M) alone had no effect in the absence of HGF, but significantly increased p42 MAPK induction by HGF. Moreover, the cell-permeable cAMP analog, 8-bromo cAMP (10(-7) M), and phorbol 12-myristate 13 acetate (10(-7) M) potentiated HGF-induced MAPK phosphorylation. The effects of these analogs were antagonized by the protein kinase A (PKA) inhibitor H-89 (10(-7) M) and the protein kinase C (PKC) inhibitor sphingosine (10(-6) M), respectively. These results suggest that direct or indirect activation of both PKA and PKC represent a positive regulatory mechanism for stimulating MAPK induction by HGF. PMID- 17409629 TI - Regulation of Cav1.2 current: interaction with intracellular molecules. AB - Ca(V)1.2 (alpha(1c)) is a pore-forming subunit of the voltage-dependent L-type calcium channel and is expressed in many tissues. The beta and alpha(2)/delta subunits are auxiliary subunits that affect the kinetics and the expression of Ca(V)1.2. In addition to the beta and alpha(2)/delta subunits, several molecules have been reported to be involved in the regulation of Ca(V)1.2 current. Calmodulin, CaBP1 (calcium-binding protein-1), CaMKII (calcium/calmodulin dependent protein kinase II), AKAPs (A-kinase anchoring proteins), phosphatases, Caveolin-3, beta(2)-adrenergic receptor, PDZ domain proteins, sorcin, SNARE proteins, synaptotagmin, CSN5, RGK family, and AHNAK1 have all been reported to interact with Ca(V)1.2 and the beta subunit. This review focuses on the effect of these molecules on Ca(V)1.2 current. PMID- 17409630 TI - Arrhythmia models for drug research: classification of antiarrhythmic drugs. AB - The aim of this study was to classify antiarrhythmic drugs based on their effectiveness on 6 in vivo arrhythmia models, mainly using dogs. The models were produced by two-stage coronary ligation, digitalis, halothane-adrenaline, programmed electrical stimulation in old myocardial infarction dogs, coronary artery occlusion/reperfusion, or chronic atrioventricular block. Na(+)-channel blocking drugs suppressed two-stage coronary ligation and digitalis arrhythmias. Ca(2+)-channel blockers and beta-blockers suppressed halothane-adrenaline arrhythmia. Positive inotropic drugs aggravated halothane-adrenaline arrhythmia, but did not aggravate digitalis arrhythmia. K(+)-channel blockers suppressed programmed electrical stimulation induced arrhythmia, but induced torsades de pointes type arrhythmia in chronic atrioventricular block dogs and aggravated halothane-adrenaline arrhythmia. Na(+)/H(+)-exchange blockers suppressed coronary artery occlusion/reperfusion arrhythmias. This classification may be useful for predicting the clinical effectiveness in the preclinical stage of drug development. PMID- 17409631 TI - Endothelin-1 induces contraction via a Syk-mediated p38 mitogen-activated protein kinase pathway in rat aortic smooth muscle. AB - Although spleen tyrosine kinase (Syk) has crucial roles in various cells, its function on vascular smooth muscle contraction has not been determined. In the present study, we performed experiments to determine if Syk contributes to the endothelin-1 (ET-1)-mediated contraction in rat aortic smooth muscle. ET-1 induced contraction of aortic strips was inhibited by piceatannol, PD98059, and SB203580, inhibitors of Syk, extracellular signal-regulated kinase 1/2 (ERK1/2), and p38 mitogen-activated protein kinase (MAPK), respectively. Piceatannol also attenuated high K(+)-induced contraction. ET-1 dose-dependently enhanced the activity of Syk and this was inhibited by piceatannol in both rat aortic strip and rat aortic smooth muscle cells. The phosphorylation of p38 MAPK and heat shock protein 27 (HSP27), but not that of ERK1/2, in response to ET-1 was inhibited by both piceatannol and SB203580. These results suggest that Syk may play an important role in the regulation of aortic smooth muscle contraction induced by ET-1, which may be mediated by the p38 MAPK/HSP27 signaling pathway. PMID- 17409632 TI - The long-acting Ca2+-channel blocker azelnidipine prevents left ventricular remodeling after myocardial infarction. AB - Long-acting Ca(2+)-channel blockers have been reported to be effective in treating ischemic heart disease. However, their effects on cardiac remodeling after myocardial infarction (MI) are still unclear. We performed this study to examine the effect of azelnidipine on left ventricular (LV) remodeling, including systolic and diastolic dysfunction, in rats with MI. MI was induced by ligation of the left anterior descending artery. The rats were then separated into 3 groups: a sham-operated group (n = 9), untreated MI group (n = 10), and azelnidipine-treated MI group (n = 10). Four weeks after MI, hemodynamic measurements and Doppler echocardiographic assessment were performed. LV weight and LV end-diastolic dimension were significantly higher in the untreated MI group than in the sham-operated group. Azelnidipine significantly prevented the increases in these parameters. Azelnidipine also improved the ejection fraction (42 +/- 3%, P<0.05) and the E wave to A wave ratio (3.2 +/- 0.5, P<0.05), compared with the untreated MI group (31 +/- 3% and 5.3 +/- 0.8, respectively). In conclusion, azelnidipine can prevent LV remodeling and improve systolic and diastolic function after MI. Administration of long-acting Ca(2+)-channel blockers after MI is an effective strategy for treating MI. PMID- 17409633 TI - Stearoyl-CoA desaturase activity is elevated by the suppression of its degradation by clofibric acid in the liver of rats. AB - A mechanism by which fibrates control stearoyl-CoA desaturase (SCD) in the liver was studied. Treatment of rats with 2-(4-chlorophenoxy)-2-methylpropionic acid (clofibric acid) or feeding of a fat-free diet markedly elevated hepatic activity of SCD. Both the treatment with clofibric acid and the feeding of the fat-free diet caused an increase in the steady-state level of SCD1 mRNA and enhanced transcriptional rate. The half-lives of SCD for control rats, rats treated with clofibric acid rats, and rats fed the fat-free diet were estimated to be 2.0, 3.9, and 1.9 h, respectively. Activity of palmitoyl-CoA chain elongase (PCE) was increased by both clofibric acid treatment and feeding of the fat-free diet as was observed with SCD. Steady-state level of rat fatty acid elongase 2 mRNA was increased by the treatment with clofibric acid or feeding of fat-free diet, although the transcriptional rate was not altered. Different from SCD, PCE was highly stable and its half-life was not changed by either clofibric acid or fat free diet. These results strongly suggest that the decreased degradation of SCD is responsible for the increase in its activity in addition to increased transcription of SCD1 in the rats treated with clofibric acid. PMID- 17409634 TI - Stimulation of histamine H1 receptor up-regulates histamine H1 receptor itself through activation of receptor gene transcription. AB - Histamine is a major mediator in allergy acting mainly through the histamine H(1) receptor (H1R). Although H1R up-regulation has been suggested as an important step for induction of allergic symptoms, little is known about the regulation of H1R level. Here we report that the activation of H1R up-regulates H1R through augmentation of H1R mRNA expression in HeLa cells. Histamine stimulation significantly increased both H1R promoter activity and mRNA level without alteration in mRNA stability. H1R protein was also up-regulated by histamine. An H1R antagonist but not histamine H(2) receptor antagonist blocked histamine induced up-regulation of both promoter activity and mRNA expression. A protein kinase C (PKC) activator, phorbol-12-myristate-13-acetate, increased H1R mRNA expression, whereas an activator of PKA or PKG (8-Br-cAMP or 8-Br-cGMP, respectively) did not. Furthermore, histamine-induced up-regulation of both promoter activity and mRNA level were completely suppressed by the PKC inhibitor Ro-31-8220. H1R antagonists have long been thought to block H1R and inhibit immediate allergy symptoms. In addition to this short-term effect, our data propose their long-term inhibitory effect against allergic diseases by suppressing PKC-mediated H1R gene transcription. This finding provides new insights into the therapeutic target of H1R antagonist in allergic diseases. PMID- 17409635 TI - Chotosan, a kampo formula, ameliorates chronic cerebral hypoperfusion-induced deficits in object recognition behaviors and central cholinergic systems in mice. AB - We previously demonstrated that the Kampo formula chotosan (CTS) ameliorated spatial cognitive impairment via central cholinergic systems in a chronic cerebral hypoperfusion (P2VO) mouse model. In this study, the object discrimination tasks were used to determine if the ameliorative effects of CTS on P2VO-induced cognitive deficits are a characteristic pharmacological profile of this formula, with the aim of clarifying the mechanisms by which CTS enhances central cholinergic function in P2VO mice. The cholinesterase inhibitor tacrine (THA) and Kampo formula saikokeishito (SKT) were used as controls. P2VO impaired object discrimination performance in the object recognition, location, and context tests. Daily administration of CTS (750 mg/kg, p.o.) and THA (2.5 mg/kg, i.p.) improved the object discrimination deficits, whereas SKT (750 mg/kg, p.o.) did not. In ex vivo assays, tacrine but not CTS or SKT inhibited cortical cholinesterase activity. P2VO reduced the mRNA expression of m(3) and m(5) muscarinic receptors and choline acetyltransferase but not that of other muscarinic receptor subtypes in the cerebral cortex. Daily administration of CTS and THA but not SKT reversed these expression changes. These results suggest that CTS and THA improve P2VO-induced cognitive impairment by normalizing the deficit of central cholinergic systems and that the beneficial effect on P2VO-induced cognitive deficits is a distinctive pharmacological characteristic of CTS. PMID- 17409636 TI - Links between de novo fatty acid synthesis and leptin secretion in bovine adipocytes. AB - Leptin secretion by adipose tissue is involved in many physiological control systems, including those that determine growth, development, body composition, milk production, and reproductive function. In the adipocyte of monogastric animals, malonyl CoA (coenzyme A) seems to link the flux of energy substrates to the control of leptin production. In this study, we tested this for ruminants by examining the effect of cerulenin, an inhibitor of de novo fatty acid synthesis at the step from malonyl CoA to palmitate, on leptin production by cultured bovine adipocytes derived from intermuscular fat. Purified preadipocytes were obtained by the ceiling culture method, and adipogenic media were used to induce their differentiation into adipocytes. We found that leptin concentrations increased significantly with time in culture, and with increases in glucose concentration. Addition of 2-deoxy-D-glucose to the medium, a competitive inhibitor of glucose transport and metabolism, suppressed leptin secretion. In media with high glucose concentrations, cerulenin enhanced leptin secretion. We conclude that, as in monogastrics, malonyl CoA may play a key role in the control of leptin secretion in ruminants. PMID- 17409637 TI - Immunohistochemical studies on cytokeratin 8 and 18 expressions in canine cutaneous adnexus and their tumors. AB - The expressions of cytokeratin 8 and 18 (CK8 and CK18) in the normal canine skin (2 cases) and cutaneous adnexal tumors (127 cases) were investigated immunohistochemically. In the normal skin, co-expression of CK8/18 was found in the glandular epithelium of apocrine sweat glands, and single CK8 immunoreactivity was detected occasionally in the external root sheath at the isthmus and suprabulbar regions of the hair follicles. Neoplastic glandular epithelial cells in all apocrine gland tumors (21/21 cases, 100%) had co expression of CK8/18. In trichoblastomas (27/28 cases, 96%), most neoplastic cells were diffusely positive for CK8, but those were negative for CK18. Single CK8-expression was also observed in basaloid neoplastic cells in several cases of trichoepitheliomas (7/19 cases, 37%) and pilomatricoma (1/7 cases, 14%). In several cases of trichoblastomas (4/28 cases, 14%) and trichoepitheliomas (2/19 cases, 11%), tumor cells forming glandular structures had co-expression of CK8/18. There were no positive reactions for both CK8 and 18 in infundibular keratinizing acanthomas, and sebaceous and hepatoid gland tumors. The present findings indicate that co-expression of CK8/18 is a specific feature of apocrine sweat glands and single CK8-expression represents the natures of external root sheath or pluripotential stem cells. Thus, the combination of CK8- and 18 immunostainings may have the utility to confirm the directions of differentiation in canine cutaneous adnexal tumors providing a reliable hallmark for histopathological diagnoses. PMID- 17409638 TI - Autoantibodies against glial fibrillary acidic protein (GFAP) in cerebrospinal fluids from Pug dogs with necrotizing meningoencephalitis. AB - Cerebrospinal fluids (CSFs) from 9 Pug dogs with necrotizing meningoencephalitis (NME: Pug dog encephalitis) were examined to identify the antigens for anti astrocyte autoantibodies. Each CSF exhibited a positive reaction to the cytoplasm of cultured canine astrocytes by an indirect fluorescent antibody test. In an immunoblotting analysis on normal canine brain proteins, eight of 9 CSFs showed a common band of 52 kDa, corresponding to glial fibrillary acidic protein (GFAP), and all of 9 CSFs reacted with purified bovine GFAP. From these results, GFAP is one of the common autoantigens in Pug dogs with NME. On the other hand, the reactivity of CSFs to chymotrypsin-digested bovine GFAP fragments were variable among dogs, indicating that the antibodies in the CSFs recognized different epitopes on GFAP. PMID- 17409639 TI - Effects of low protein intake on the development of the remaining kidney in subtotally nephrectomized immature rats: apoptosis and epidermal growth factor. AB - Effects of low protein intake on the development of the remaining kidney in subtotally (5/6) nephrectomized immature rats were examined. Three week-old weaning rats were kept on a diet containing either 12% (Lp rats) or 18% (Np rats) protein for 4 or 8 weeks after subtotal nephrectomy. Blood urea nitrogen (BUN) concentration was determined at 2, 4, 6, and 8 weeks after 5/6 nephrectomy. At 4 or 8 weeks after the operation, glomerular sclerosis and tubulointerstitial damage were assessed by a standard semiquantitative analysis and were expressed as the glomerular sclerosis index (GSI) and interstitial fibrosis score (IFS), respectively. The localization of DNA fragmented cells in the kidney was examined by the terminal deoxynucleotidyl transferase (TdT) -mediated d-UTP-biotin nick end labeling (TUNEL) method and the localization of the epidermal growth factor (EGF) by immunohistochemical methods. BUN concentration was significantly lower in the Lp rats compared with that in the Np rats. Both 4 and 8 weeks after subtotal nephrectomy, GSI and incidence of TUNEL positive cells in the distal tubules were significantly lower in the Lp rats than in the Np rats. Four weeks after the operation, IFS was significantly lower in the Lp rats than in the Np rats. Four and 8 weeks after the operation, EGF positive cells in the distal tubules were more observed in the Lp rats than in the Np rats. These findings reveal that protein restriction is effective in preventing renal tubular scarring in immature rats and that EGF is involved in the process of this prevention. PMID- 17409640 TI - Chondrocyte apoptosis and decrease of glycosaminoglycan in cranial cruciate ligament insertion after resection in rabbits. AB - We examined time-dependent histological changes of the calcified fibrocartilage area in a tibial cranial cruciate ligament (CCL) insertion after ligament resection in rabbits. The animals were divided into two groups: those undergoing CCL substance resection in the right stifle (resected group) and those receiving the same operation without CCL resection in the left stifle (sham operated group). Five animals were euthanized with deep anaesthesia at four time periods (1, 2, 4 and 6 weeks), and Haematoxylin-eosin and Safranin-O stainings and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) staining were performed. The average percentage of TUNEL-positive chondrocytes and the average thickness of the glycosaminoglycan (GAG)-stained area in the calcified fibrocartilage area were measured. Two and 4 weeks after the surgery, the average percentages of TUNEL-positive chondrocytes in the resected group (23.8 +/- 10.3% and 15.9 +/- 6.7%, respectively) were significantly higher than those in the sham operated group (8.9 +/- 3.8% and 7.4 +/- 1.6%, P<0.05, respectively). Six weeks after the surgery, the average thickness of the GAG-stained area in the resected group (7.7 +/- 13. 5 microm) was significantly smaller than that in the sham operated group (69.4 +/- 39.9 microm, P<0.05). Our results suggest that the average percentage of TUNEL positive chondrocytes became a peak in 2 weeks and that histological changes occurred in 6 weeks. The chondrocyte apoptosis can induce decrease of GAG-stained area after resection of CCL. Therefore, chondrocyte apoptosis in the calcified cartilage area in the CCL tibial insertion might lead to histological changes. PMID- 17409641 TI - Serological evidence of influenza A virus infection in Kuril harbor seals (Phoca vitulina stejnegeri) of Hokkaido, Japan. AB - For proper management and conservation of the Kuril harbor seal (Phoca vitulina stejnegeri) through disease control, serological analysis was performed for influenza A virus infection in free-ranging seals in Hokkaido, Japan. Serum samples were collected from seals at Nosappu (231 seals), Akkeshi (16) and Erimo (75), between 1998 and 2005, and were analyzed by ELISA. Antibodies to the influenza A virus were detected only in seals from Nosappu. The incidences were 11% (1/9), 3% (2/66), 12% (7/59) and 6% (5/77) in 1998, 2003, 2004 and 2005, respectively. These suggest sporadic infection. Because antibody-positive seals included juvenile seals in each year, the infections were considered to have been circulated since no later than the late 1990s until recent years. ELISA-positive sera were analyzed by hemagglutination inhibition (HI) tests to determine the subtypes. Antibodies to the H3 and H6 subtypes were detected in 10 and 2 sera, respectively. Two of the sera that had antibodies to the H6 subtype also had antibodies to the H3 subtype. These two seals were considered to have been infected with both the H3 and H6 subtypes. This is the first investigation to find antibodies to the H6 subtype in seals. Although the H6 subtype had been isolated only from avians, genetic analysis had suggested that the H6 subtype could become a novel mammalian pathogen. For definitive diagnosis, detection of the virus from the tissue or mucus of seals is required. PMID- 17409642 TI - Biomarkers for the activation of calcium metabolism in dairy cows: elevation of tartrate-resistant acid phosphatase activity by lowering dietary cation-anion difference is associated with the prevention of milk fever. AB - In our previous study, it was demonstrated that the administration of anion salts, which slightly lower the dietary cation-anion difference (DCAD), in the prepartum period is safe and effective for preventing milk fever in multiparous cows. In the present study, several biomarkers, which might show activation of Ca metabolism, were analyzed using stored samples in the previous study to investigate the mechanism of the preventive effect on milk fever by lowering DCAD. Changes in bone-specific alkaline phosphatase activity, osteocalcin and insulin-like growth factor I concentrations in serum were almost the same among the three groups of multiparous cows with or without the oral administration of anion salts, while the levels of these serum biomarkers in the group of primiparous cows (heifer group) were much higher compared with those in the three multiparous groups throughout the experimental period. Urinary deoxypyridinoline excretion was not a useful biomarker for dairy cows because it hardly changed during the peripartum period in all groups. However, serum tartrate-resistant acid phosphatase (TRAP) activity, which is known as a biomarker of osteoclast activity, was well associated with the administration of anion salts lowering DCAD because among the three multiparous groups, only the group of multiparous cows fed the anion salts (anion group) showed an increased level, which rose to the level in the heifer group, and was markedly higher than those in the other control groups of multiparous cows. The increased activity of serum TRAP in the anion group suggested that Ca in the plasma pool was mobilized smoothly from bone bound Ca via mature osteoclasts at parturition, which might be due to prior activation under mild acidosis induced by slightly lowering DCAD. Therefore, TRAP was the best biomarker to monitor the activation of Ca metabolism in dairy cows fed anion salts. PMID- 17409643 TI - The significance of p53 and retinoblastoma pathways in canine hemangiosarcoma. AB - To investigate whether inactivation of the p53 and retinoblastoma (Rb) protein pathways contributes to the development of canine hemangiosarcoma (HSA), we examined immunohistochemically the expression of p53, Rb, phosphorylated Rb (phospho-Rb), p16, and cyclin D1 in 39 spontaneous canine HSAs and 10 hemangiomas. In addition, mutations in the p53 gene were analyzed by polymerase chain reaction (PCR)-single-stranded conformation polymorphism and PCR direct sequencing; furthermore, we quantified cyclin D1 mRNA by semiquantitative real time reverse transcription-PCR. Positive immunoreactivity for p53 was observed in 17.9% of HSAs. However, mutations were not detected in these cases. The labeling indices for Rb, phospho-Rb, and cyclin D1 were markedly higher in all HSAs than in hemangiomas. Of the 7 cases with cyclin D1-positive immunoreactivity, 4 overexpressed cyclin D1 mRNA (to a level more than 10-fold higher than that of GAPDH mRNA). The p16 protein was clearly detected in all hemangiomas; however, 82% of the neoplastic cells in HSA showed a loss of or low immunoreactivity. These results suggest that alteration of the p16-cyclin D1-Rb pathway, rather than the p53 pathway, may be associated with the pathogenesis of canine HSA. PMID- 17409644 TI - Role of NF-kappaB in constitutive expression of MAIL in epidermal keratinocytes. AB - Molecule possessing ankyrin-repeats induced by lipopolysaccharide (MAIL) is a nuclear IkappaB protein that is also known as interleukin-1-inducible nuclear ankyrin repeat protein and inhibitor of nuclear factor kappaBzeta (IkappaBzeta). We previously observed that MAIL-deficient mice were affected by atopic dermatitis-like skin lesions and demonstrated the importance of MAIL in the skin. In this study, we investigated MAIL expression in mouse keratinocytes. MAIL mRNA was constitutively expressed in the skin epidermis. MAIL expression was also confirmed in primary keratinocytes and the PAM212 keratinocyte cell line. The inhibitors of nuclear factor kappaB (NF-kappaB)-Bay11-7082 and the IkappaBalphaM supersuppressor-considerably downregulated MAIL expression in the keratinocytes. Immunoreactivity for NF-kappaB components was localized in the cytoplasm and nucleus of normal unstimulated keratinocytes. The expression level of MAIL in the skin did not change following lipopolysaccharide (LPS) administration to mice. Interestingly, in accordance with the in vivo findings, the MAIL expression level did not change following LPS stimulation even in primary keratinocytes; however, MAIL expression was strongly increased by interleukin-1 stimulation. These results collectively suggest that the constitutive expression of MAIL in keratinocytes is controlled, at least in part, by NF-kappaB and that there may be LPS-specific repressive mechanisms that inhibit MAIL induction. PMID- 17409645 TI - 17Beta-estradiol prevents the glutamate-induced decrease of Akt and its downstream targets in HT22 cells. AB - Estradiol is known to exert neuroprotective effect against glutamate toxicity in hippocampal-derived cell line (HT22). This study investigated whether estradiol modulates the anti-apoptotic signal through the phosphorylation of Akt and its downstream targets, including Bad, forkhead transcription factors FKHR and FKHRL1. Pretreatment with 17beta-estradiol decreased glutamate toxicity-induced cell death in HT22 cells. Also, pretreatment with 17beta-estradiol significantly decreased the positive cells of TUNEL stain, compared to that of only glutamate treated cells. Potential activation was measured by phosphorylation of Akt at Ser(473), Bad at Ser(136), FKHR at Ser(256), and FKHRL1 at Thr(32) using Western blot analysis. 17Beta-estradiol pretreatment prevented the glutamate-induced decrease of pAkt, pBad, pFKHR, and pFKHRL1. These findings clearly confirm that 17beta-estradiol plays a potent neuroprotective role against glutamate-induced toxicity and suggest that phosphorylation of Akt and its downstream targets by 17beta-estradiol mediated these protective effects. PMID- 17409646 TI - A new method of measuring the transepidermal water loss (TEWL) of dog skin. AB - Human skin barrier function is evaluated by measuring transepidermal water loss (TEWL). However, this conventional method has not been applied to assess canine skin barrier function because the equipment is not suitable for dogs due to the effects of air turbulence resulting from movement of the subject and vapor from the subject's hair coat. The TEWL analyzer CC-01 was developed as a closed chamber method device; this means that instead of using the open-chamber method, it has a ventilated chamber that uses dry air. TEWL values measured by CC-01 show less variability than those measured by the conventional method. An ambient temperature of 20-26 degrees C is optimal for measurement with the CC-01, and humidity affects the length of measurement but not the values. The CC-01 may be more reliable for measurement of TEWL than the conventional methods and may give new insights in the evaluation of skin barrier function in dogs. PMID- 17409647 TI - Clinical pharmacokinetics of oseltamivir and its active metabolite oseltamivir carboxylate after oral administration in horses. AB - The aim of this study was to investigate the pharmacokinetics of oseltamivir carboxylate (OC) in horses (n=6) after oral administration of its prodrug oseltamivir. The binding rate of OC to horse plasma proteins was negligible (<1%). Oral administration of oseltamivir of 2 mg/kg body weight of oseltamivir to horses provided a plasma concentration of OC (mean maximum concentration: 257.9 ng/ml) above the inhibitory concentrations against equine influenza A viruses determined in vitro. However, because OC is rapidly eliminated from horse plasma (mean elimination half-life: 2.5 hr), administration intervals should be less than 10 hr to retain a suitable concentration when using a single dose of 2 mg/kg oseltamivir. PMID- 17409648 TI - Multiple osseous metastases of a carotid body tumor in a dog. AB - Metastasis of malignant carotid body tumor to multiple bones was detected in a 13 year-old female Siberian husky dog. Radiographs exhibited an abnormal mass in the retropharyngeal site and osteolytic lesions in the vertebral bodies, spinous process, tibia, and ribs. At necropsy, multiple masses were observed in the bones as well as at the dorsal area of the retropharynx. Histologically, the tumor cells, arranged in sheets and clusters, had eosinophilic finely granular cytoplasm. Immunohistochemistry showed the tumor cells were positive for neuron specific enolase and synaptophysin. Electron microscopy demonstrated a number of dense membrane-bound granules in the cytoplasm of the tumor cells. Based on these findings, this case was diagnosed as multiple bone metastases of a malignant carotid body tumor. Spinal cord damage induced by the tumor mass was the cause of the hind limb paralysis of the present dog. PMID- 17409649 TI - The infectivity and pathogenicity of a group 2 bovine coronavirus in pups. AB - Canine respiratory coronavirus (CRCoV), which is more closely related to the bovine coronavirus (BCoV), has recently been detected in dogs. In this study, we examined whether BCoV was capable of infecting and exhibiting pathogenicity in dogs. Three 1-month-old pups were oronasally given field isolates of BCoV, and were kept together with 2 control animals. As a result, increases in BCoV neutralizing antibody titers were confirmed in all pups in the challenged and control groups. Moreover, the virus gene was also detected in oral and rectal swabs by RT-PCR. These results indicate that BCoV infects dogs, and easily infects other dogs that are kept together. However, no clinical symptoms such as respiratory symptoms and diarrhea were observed. PMID- 17409650 TI - A direct enzyme immunoassay for the measurement of furosemide in horse plasma. AB - A new enzyme immunoassay (EIA) for the measurement of furosemide in horse plasma is described. The lower limit of detection of this EIA method was 7.8 ng/ml. The intra-and inter-assay coefficients of variation ranged from 2.5% to 4.9% and 7.5% to 9.8%, respectively. Cross-reactivity with other compounds was not observed. There was a high correlation (r2=0.987) between the high-performance liquid chromatography and EIA results obtained for furosemide concentrations in horse plasma. These results indicate that the newly developed EIA method is useful for the quantitative analysis of furosemide in horse plasma. PMID- 17409651 TI - Salmonella isolates from cloacal swabs and footpads of wild birds in the immediate environment of Tokyo Bay. AB - A total of 328 cloacal swabs and 163 footpads of wild birds were investigated for the presence of salmonellae. All 19 isolates from cloacal swabs were serotyped as Salmonella Typhimurium susceptible to all five conventional antimicrobial agents (ampicillin, chloramphenicol, streptomycin, oxytetracycline and nalidixic acid) tested. In contrast, 15 salmonellae isolated from footpads included S. Muenhen, S. Virchow, S. Bareily and S. Bovismorbificans, including S. Typhimurium; these non-Salmonella Typhimurium isolates showed multiple drug resistance. PMID- 17409652 TI - Japanese black cattle with orotic aciduria detected by gas-chromatography/mass spectrometry. AB - A 4-months-old calf of Japanese black cattle was diagnosed with orotic aciduria by gas-chromatography/mass-spectrometry (GC/MS). Until now orotic aciduria had not been reported in Japanese black cattle. The animal showed repeated diarrhea. The hematocrit was low, and microcytes and acanthocytes were observed in blood smears. The calf had lower serum total protein concentrations with a higher blood ammonia concentration. Needle-shaped crystals of orotic acid were observed in urinary sediments. Sequence homologous analysis with cattle uridine monophosphate synthase DNA indicated silent mutation in the affected calf. PMID- 17409653 TI - Immunohistochemical examination of PDGF-AB, TGF-beta and their receptors in the hemocytes of a tick, Ornithodoros moubata (Acari: Argasidae). AB - Growth factors, Platelet Derived Growth Factor (PDGF) and Transforming Growth Factor (TGF)-beta, were demonstrated in vertebrate and invertebrate immmunocytes. It is generally known that the growth factors are important in various biological processes, such as the regulation of cell differentiation, development and wound healing. In the present study, the presence of TGF-beta1 and PDGF-receptor-alpha in plasmatocytes and PDGF-AB in granulocytes of a soft tick, Ornithodoros moubata, was confirmed immunohistochemically. The tick midgut might be damaged by intracellular digestion and penetration of protozoa. Therefore, it is considered that PDGF from granulocytes may affect the PDGF-receptor-alpha in plasmatocytes and TGF-beta from plasmatocytes may function to repair the midgut. The results obtained here add to the elucidation of the functions of tick hemocytes. PMID- 17409654 TI - Serum amyloid A and haptoglobin levels in bovine amyloidosis. AB - Serum amyloid A (SAA) and haptoglobin (Hp) levels were determined in 25 cows suffering from amyloidosis. SAA levels in cows with amyloidosis ranged between < 0.3 and 225.8 microg/ml, with a median level of 105.1 microg/ml, and Hp levels ranged between < 20 and 1860 microg/ml, with a median level of 950 microg/ml. These levels were significantly higher than the levels observed in healthy cows (SAA levels ranged from < 0.3 to 13.5 microg/ml, with median of 1.4 microg/ml, and Hp levels were undetectable in all cases), but were not significantly different from the levels observed in control cows with chronic inflammation. There was a significant correlation between SAA and Hp levels in cows with chronic inflammation , but not in cows with amyloidosis. It was concluded that the serum SAA levels in cows with amyloidosis might be changed by some factor other than inflammation. PMID- 17409655 TI - Aspiration pneumonia as a cause of neonatal death in three captive bottlenose dolphins (Tursiops truncates). AB - Neonatal weakness of calves is one of the common reproductive-related problems in captive cetaceans; clinical signs can be observed in the first few hours after delivery. Three 3-day-old bottlenose dolphins died with history of weakness since birth. Pathological study demonstrated purulent bronchopneumonia associated with prominent bacterial colonies and foreign substances in alveoli, suggesting aspiration pneumonia as a cause of neonatal weakness and resultant death of the three calves. PMID- 17409656 TI - [Molecular basis of metabolic syndrome]. PMID- 17409657 TI - [Dramatic increase of fatty liver in recent fifteen years probably due to changes of Japanese life-style]. PMID- 17409658 TI - [Changing trends of acid related diseases in Japan and westernization of diet]. PMID- 17409659 TI - [Colorectal cancer and lifestyle, lifestyle related diseases]. PMID- 17409660 TI - [Relations of metabolic syndrome and bright pancreas]. PMID- 17409662 TI - [A case of carcinosarcoma of the esophagus treated by the chemoradiotherapy]. AB - A 82-year-old man was admitted into our hospital complaining of dysphagia. Endoscopic examination revealed an elevated tumor in the middle esophagus, and biopsy revealed carcinosarcoma. He underwent chemo-radiotherapy, and the size of tumor decreased remarkably and almost disappeared 7 months later. However, a metastatic lesion appeared 11 months later, and eventually he died. Autopsy revealed a sarcomatous element in the metastatic part, but the primary lesion showed no recurrence. We report herein this rare case of carcinosarcoma of the esophagus that was treated with chemo-radiotherapy. PMID- 17409663 TI - [A case of gastroesophageal reflux disease with marked eosinophilia]. AB - A 73-year-old woman without a history of allergic diseases visited our hospital complaining of sore throat and nocturnal cough. Blood tests showed marked eosinophilia (18000/mm(3);WBC 21900/mm(3), Eos 82.0%) with normal serum levels of C-reactive protein, non-specific and various allergen-specific IgE. Stool tests for protozoa or helminthic ova were negative. Chest X-ray films showed no pulmonary abnormalities. Endoscopic and histological examinations revealed reflux esophagitis (grade C according to the Los Angeles Classification System) with hiatal hernia with inflammatory infiltrates including eosinophils within the esophageal mucosa. A computed tomography showed the thickening of the esophageal wall. An administration of lansoprazole improved reflux esophagitis and also eosinophilia, and an alteration to famotidine caused heartburn with an increase in eosinophils. A re-alteration to omeprazole relieved the symptom and decreased eosinophils. It was shown that gastroesophageal reflux disease was one of the possible causes of eosinophilia. PMID- 17409664 TI - [A case of secondary gastric wall abscess due to perforation from advanced colonic cancer successfully treated by endoscopic endoprostesis using endoscopic retrograde biliary drainage tube]. AB - A 44 year-old man was admitted to our hospital because of severe left upper abdominal pain and high fever. Blood examinations revealed high levels of white blood cell count and CRP. An abdominal enhanced CT scan showed a low-density mass with an enhanced margin at the posterior side of the gastric body, and an irregular mass nearby at the transverse colon. We diagnosed a gastric wall abscess due to perforation from advanced colonic cancer and performed endoscopic drainage and endoprostesis by ERBD tube insertion into the abscess through the gastric wall. After the procedure, the clinical symptoms and laboratory data improved quickly, and we could continue further examinations and proceed safety with the treatment plan. To our knowledge, this is the first reported case of a gastric wall abscess due to penetration by gastrointestinal tract cancer which was treated with endoscopic endoprostesis. PMID- 17409665 TI - [A case of nonocclusive mesenteric ischemia (NOMI) which occurred in a patient with torsion of gallbladder]. AB - We report a case of nonocclusive mesenteric ischemia (NOMI) which occurred in a patient with torsion of gallbladder. A 91-year-old woman was admitted to the hospital and was diagnosed of acute cholecystitis. The next day, she went into shock. Then, we diagnosed her illness as torsion of gallbladder by computed tomography and ultrasonography, and performed an emergency operation. After cholecystectomy, it was recognized that the wide range of the small intestine had become necrotic sporadically. We diagnosed it as NOMI, and performed the wide resection of the small intestine followed by making double stomas. There is no previous report of NOMI associated with torsion of gallbladder. We guess the cause of NOMI in this case would be dehydration because of gallbladder torsion. NOMI has high mortality. Early diagnosis and early treatment are of great importance in NOMI. PMID- 17409666 TI - [A case of ruptured ileal varices associated with alcoholic liver cirrhosis]. AB - The case was a 33-year-old woman with alcoholic liver cirrhosis. Her chief complaint was melena. She had undergone the abdominal operation in the past. Upper and lower gastrointestinal endoscopy did not reveal the bleeding point in the gastrointestinal tract. We diagnosed the bleeding from the ileocecum by the hemorrhage scintigraphy, the superior mesenteric artery angiography, and CT angiography. She had varices at the terminal ileum and adhesion between the ovary and the ileum end. The subtotal resection of the ileum was performed. PMID- 17409667 TI - [A case of hepatocellar carcinoma associated with mixed connective tissue disease]. AB - A 79-year-old woman, who had been treated because of mixed connective tissue disease (MCTD), was admitted to the hospital for the intensive examination of a hepatic tumor that was unexpectedly found on computed tomography. The hepatic tumor was approximately 40 mm in diameter. Her transaminase levels were slightly elevated but hepatic virus markers were negative. However, a liver aspiration biopsy revealed moderately differentiated hepatocellular carcinoma (HCC). In the surrounding non-tumor area, nonspecific reactive hepatitis was detected. MCTD associated with HCC has not yet been reported in the past. This case appears to be very interesting in terms of the etiology of HCC. PMID- 17409668 TI - [A case of intractable chronic abdominal pain caused by hepatomegaly associated with primary amyloidosis, successfully treated with celiac plexus block]. AB - A 51-year-old female previously diagnosed as primary amyloidosis suffered from recurrent abdominal pain. The result of thorough examination indicated that the main cause of the pain was severe hepatomegaly. Continuous venous administration of narcotics and other alternative therapies did not provide symptomatic relief, and thus the patient was treated with celiac plexus block, which resulted in effective pain control and improved ADL level. Though the procedure of celiac plexus block is simple and celiac plexus block is applicable without causing severe complication, it is not widely used. From this case, it is considered that celiac plexus block is one of the most effective means to relieve intractable pain associated with both benign malady and abdominal malignant tumor. PMID- 17409669 TI - [A case of histiocytic necrotizing lymphadenitis showing focal lymph nodes swelling in the abdomen]. AB - A case was 23 years old woman. She came to our hospital with complain of continuing fever after common cold and mass like an egg was touched in right lower abdomen. No other surface lymph node was swelling. Abdominal US, CT and MRI showed that the mass was existed in the front of the psoas muscle and the maximum diameter of it was 6 cm in which necrotizing lesion was recognized. The size of the lymph node became smaller slowly but we could not perfectly exclude malignant disease, we examined needle biopsy for the diagnosis. The histopathological findings showed the histiocytic necrotizing lymphadenitis (HNL). HNL usually appears in the neck area, but in this case it was detected only in the abdomen. This case was very rare and we reported. PMID- 17409670 TI - Recent progress in the treatment of pulmonary arterial hypertension: expectation for rho-kinase inhibitors. AB - Pulmonary arterial hypertension (PAH) is a disease with poor prognosis characterized by progressive elevation of pulmonary arterial pressure and vascular resistance due to pulmonary artery hyperconstriction and remodeling. However, the precise mechanism of PAH still remains to be elucidated. Although anticoagulant agents, vasodilators (e.g., prostaglandins, sildenafil, and bosentan), and lung transplantation are currently used for the treatment of PAH, more effective treatment needs to be developed. Rho-kinase causes vascular smooth muscle hyperconstriction and vascular remodeling through inhibition of myosin phosphatase and activation of its downstream effectors. In a series of experimental and clinical studies, we have demonstrated that Rho-kinase-mediated pathway plays an important role in various cellular functions, not only in vascular smooth muscle hyperconstriction but also in actin cytoskeleton organization, cell adhesion and motility, cytokinesis, and gene expression, all of which may be involved in the pathogenesis of arteriosclerosis. We also have recently demonstrated that Rho-kinase is activated in animal models of PAH with different etiologies (monocrotaline and chronic hypoxia) associated with enhanced pulmonary vasoconstricting and proliferating responses, impaired endothelial vasodilator functions, and pulmonary remodeling. Indeed, we were able to demonstrate that intravenous fasudil, a selective Rho-kinase inhibitor, exerts acute pulmonary vasodilator effects in patients with severe PAH who were refractory to conventional therapies. Taken together, our findings indicate that Rho-kinase is a novel and important therapeutic target of PAH in humans and that Rho-kinase inhibitors are a promising new class of drugs for the fatal disorder. PMID- 17409671 TI - Reliability and validity of the Korean version of Kidney Disease Quality of Life instrument (KDQOL-SF). AB - Although the Kidney Disease Quality of Life instrument (KDQOL-SF), which was designed to measure the comprehensive quality of life in patients with end stage renal disease, has been tested and widely administered in many other counties, it has not yet been translated, validated and reported for Korea. The primary purpose of this study was to validate a Korean version of KDQOL-SF and to evaluate its psychometric properties. The study subjects were 164 patients with hemodialysis or continuous ambulatory peritoneal dialysis in university dialysis centers in Korea. In order to investigate the reliability, test-retest reliability and internal consistency reliability were assessed. Both test-retest reliability and internal consistency reliability were found to be high. In order to investigate the construct validation, all the items of the SF-36 scales, an established generic QOL measures, were compared with an overall health rating scale in kidney disease-targeted scale. They found to be highly correlated with each other. Moreover, overall health rating scale was significantly correlated with symptoms/problems, effects of kidney disease, burden of kidney disease, cognitive function (p < 0.001), and quality of social interaction (p < 0.05). These results suggest that the Korean version of KDQOL-SF satisfies its reliability and validity in Korean patients with hemodialysis or continuous ambulatory peritoneal dialysis. This questionnaire provides important and clinically valuable information for understanding the health related quality of life in the Korean patients with dialysis. PMID- 17409672 TI - The risk factors affecting the complications of saphenous vein graft harvesting in aortocoronary bypass surgery. AB - Problems of wound healing are commonly observed after coronary bypass grafting (CABG) operations. Our aim is to determine the prevalence and the predictors of saphenous vein harvesting complications after coronary artery bypass surgery. One hundred twenty six patients operated in the specified period were included in this prospective study. In an early period 3 patients were excluded because of their mortality. Twenty patients were female and 103 were male. Forty three patients underwent an open procedure with one incision (35%), 61 patients also underwent an open procedure but with multiple incisions (49%), and 19 patients underwent a closed procedure with stripper (16%). Complications related with leg incisions after surgery were investigated. Multiple incision technique has the longest (49.28 +/- 14.7 cm; p < 0.001) total incision length (compare to single incision and stripper technique). As incision length increases, the incidence of drainage (p < 0.01), pain score (p < 0.05), hematoma (p < 0.05) and diffuse ecchymosis (p < 0.05) were increased. Drainage was seen more frequently in female (p < 0.001) and diabetic patients (p < 0.05). Sex (p < 0.001) and incision length (p < 0.05) have been found independent risk factors for drainage complication. Superficial infection (p < 0.05), pain (p < 0.05) and dehiscence (p < 0.05) were significantly higher in female patients. As the incision length of the multiple incision technique became longer, the risk of drainage, pain, hematoma and diffuse ecchymosis were increased. The significantly increased risk for wound complications were also seen in female gender, diabetic and obese patients. PMID- 17409673 TI - Exercise training fails to modify arterial baroreflex sensitivity in ovariectomized female rats. AB - In men, exercise training attenuates age-related reduction in baroreflex sensitivity, which is related to cardiovascular health. It is unknown, however, if this holds true for post-menopausal women. We examined the effects of exercise training on baroreceptor-heart rate (HR) reflex sensitivity in ovariectomized (OVX) and sham-operated (SO) Wistar-Kyoto rats. At the age of 8 weeks, OVX and SO rats were assigned to either sedentary or exercise-trained group. Exercise training was performed on a treadmill 5 days per week. At the age of 20 weeks, baroreflex sensitivity in response to increases in blood pressure (BRSinc) and decreases in blood pressure (BRSdec) were evaluated by injections of phenylephrine and sodium nitroprusside, respectively. Both BRSinc and BRSdec were significantly reduced in sedentary OVX rats compared with sedentary SO rats. Exercise training decreased resting HR and BRSdec, but had no effect on BRSinc in SO rats. In OVX rats, exercise training decreased resting HR but modified neither BRSdec nor BRSinc. We conclude that withdrawal of female sex hormones in normotensive female rats is associated with reduced baroreflex sensitivity in response to both increase and decrease in blood pressure and that exercise training fails to modulate the decline of BRSinc associated with withdrawal of female sex hormones. To maintain high level of BRSinc in post-menopausal women, hormone replacement therapy may be needed. PMID- 17409674 TI - High serum levels of adiponectin improve coronary collateral development in patients with coronary artery disease. AB - Adiponectin is a novel polypeptide that modulates endothelial function. Association between high serum adiponectin level and stimulation of new blood vessel formation have been reported in two experimental studies, however, data in humans are lacking. We sought to determine relationship between serum adiponectin and collateral vessel development in patients with coronary artery disease. We included 89 patients with stable angina pectoris and angiographically documented total occlusion in one of the major coronary arteries. Coronary collateral circulation was graded according to Rentrop scoring method in which collateralisation was graded between 0 and 3 from the poorest to the best. Adiponectin was determined by the ELISA method. High serum adiponectin level were significantly associated with increased new collateralisation (p = 0.001). With the increase of body mass index and waist circumference, the collateral development decreased (p = 0.001, p = 0.002; respectively). Presence of the diabetes mellitus (DM) was more frequent in those with poor collateral group than in those with good collateral group. DM was associated with poor collateral development (p = 0.002). In multiple stepwise logistic regression analysis, low level of serum adiponectin (p = 0.0001), waist circumference (p = 0.001), and presence of DM (p = 0.003) were found to be significant independent predictors of poor collateral formation. In conclusion, we have shown for the first time that elevated levels of serum adiponectin are associated with coronary collateral development in patients with coronary artery disease and additionally, the present study confirms many of the metabolic associations reported previously with adiponectin. PMID- 17409675 TI - High incidence of Pertussis among schoolchildren with prolonged cough in Turkey. AB - Older children, adolescents and adults with prolonged cough should be screened for pertussis, because they can be a major source of transmission for infants who are not yet fully immunized. This study aimed to estimate the incidence of pertussis among schoolchildren of 6-14 years old with prolonged cough attending two government primary schools in central Ankara. A questionnaire and an informed consent form were sent to the parents of 1,859 schoolchildren. The former comprised questions regarding sociodemographics, and the presence and duration of cough. Parents of 1,698 (91.3%) schoolchildren returned the questionnaire. Three hundred and eighty three (22.6%) of the schoolchildren had cough for more than two weeks, and 307 (80.2%) of them, whose parents gave consent, were included in the study. Their blood samples were collected twice, with a two-week interval. Anti-pertussis toxin IgG levels were measured by enzyme-linked immunosorbent assay (ELISA). Fifty-one children (16.6%) had evidence of recent pertussis infection documented by either a titer > or = 100 ELISA Unit (EU)/ml in one of two serum samples or seroconversion in paired sera. None of the schoolchildren who had attended health care facilities for cough had been diagnosed as pertussis by a physician. In conclusion, pertussis is evident among older children and adolescents in Turkey, and in order to control the disease, priority should be given to strengthen primary immunization services along with the implementation of booster vaccinations beyond childhood. PMID- 17409676 TI - Development of a novel ELISA for detection of anti-A and anti-B antibodies in recipients of ABO-incompatible living donor liver grafts. AB - The survival rate in ABO-incompatible (ABO-I) liver transplantation was much lower than that in ABO-compatible recipients for the early experiences. It is therefore essential to develop the precise and fast measurement of anti-A and anti-B antibodies (Abs) to prevent humoral rejection in ABO-I liver transplantation. Agglutination titer has been the standard method to measure these Abs, but the interpretation of the results is subject to bias. Here, we have developed an objective and quantitative enzyme-linked immunosorbent assay (ELISA) to measure anti-A and anti-B Abs. This test requires only a small amount (10 microl) of recipient's serum. We applied the newly developed ELISA to monitor living donor liver transplant recipients and investigated the correlation between ELISA and agglutination titer. The Spearman's correlation coefficient for Abs ranged from 0.461 to 0.812. Moreover, in one case of humoral rejection, the increase of Abs was detected by ELISA one day earlier than by the agglutination titer. In conclusion, our ELISA method proved useful to detect an increase of anti-A and anti-B Abs titers at an early stage, thereby contributing to a prompt treatment of humoral rejection due to ABO-I. PMID- 17409677 TI - Independent association of high serum uric acid concentration with angiographically defined coronary artery disease. AB - Epidemiological studies have shown that a high serum uric acid concentration is a risk factor for coronary artery disease (CAD). However, the issue of whether it is an independent cardiovascular risk factor or simply a marker of co-existing conditions is a matter of controversy. In the present case-controlled study, we explored the association between serum uric acid and angiographically defined CAD in middle-aged subjects (356 CAD patients and 350 healthy individuals). Serum uric acid in CAD patients was significantly higher than that in healthy individuals (359 +/- 88.7 and 289 +/- 79.3 micromol/l, respectively, p < 0.01) and remained significantly higher after adjusting for confounding factors (F = 79.77, p < 0.01). The association between uric acid and CAD was not limited to the hyperuricemic range of values, but was also found in the high-normal range (p < 0.01). An unadjusted odds ratio (OR) of 5.0 was obtained in both genders (p < 0.01). Female patients with > 50% stenosis (clinically significant CAD), regardless of the number of diseased vessels, had higher uric acid concentrations than those with < 50% stenosis even after adjusting for confounders (F = 3.79, p = 0.01). In conclusion, we have demonstrated that high serum uric acid is independently associated with CAD and that uric acid determination could be useful as one of the markers of clinically significant CAD. PMID- 17409678 TI - On-site preparation of technetium-99m labeled human serum albumin for clinical application. AB - Technetium-99m labeled human serum albumin (Tc-99m HSA) is an important radiopharmaceutical for clinical applications, such as cardiac function tests or protein-losing gastroenteropathy assessment. However, because of transfusion induced infectious diseases, the safety of serum products is a serious concern. In this context, serum products acquired from patients themselves are the most ideal tracer. However, the development of rapid separation and easy clinical labeling methods is not yet well established. Under such situation, products from the same ethnic group or country are now recommended by the World Health Organization as an alternative preparation. This article describes the on-site preparation of Tc-99m HSA from locally supplied serum products. Different formulations were prepared and the labeling efficiency and stability were examined. Radio-labeling efficiencies were more than 90% in all preparation protocols, except for one that omitted the stannous solution. The most cost effective protocol contained HSA 0.1 mg, treated with stannous fluoride 0.2 mg, and mixed with Tc-99m pertechnetate 30 mCi. A biodistribution study was performed in rats using a gamma camera immediately after intravenous administration of radiolabeled HSA. Tissue/organ uptake was obtained by measuring the radioactivity in organs after sacrificing the rats at timed intervals. The biologic half-life was about 32 min, determined from sequential venous blood collections. These data indicate that our preparation of Tc-99m HSA is useful and potentially applicable clinically. In addition, this on-site preparation provides the possibility of labeling a patient's own serum for subsequent clinical application. PMID- 17409679 TI - Plasma interleukin-8 as a potential predictor of mortality in adult patients with severe traumatic brain injury. AB - Because of complex pathophysiology and severe consequences, traumatic brain injuries (TBI) are an important medical problem. Pathophysiology of TBI includes local and systemic stress response, in which interleukin-8 (IL-8) is considered as a key mediator of neuroinflammation. However, prognostic relevance of IL-8 measurement in adult patients with severe TBI is not certain. Therefore, IL-8 was determined in blood samples from central venous and jugular bulb catheter and in cerebrospinal fluid of twenty patients with isolated TBI at admission to Intensive Care Unit. None of the patients had history of stroke, dementia, autoimmune diseases, acute infection or medication with anti-inflammatory drugs. Ten patients died due to traumatic brain injury, while the other ten recovered well. While there was no significant difference of IL-8 levels in cerebrospinal fluid between survivors and nonsurvivors, central venous plasma level of IL-8 was significantly lower in survivors (71.00 +/- 14.17 pg/ml), than in nonsurvivors (111.26 +/- 16.9 pg/ml). Receiver Operating Characteristic (ROC) analysis revealed significant prognostic value for IL-8 in the blood as well as for the age of patients, Glasgow Coma Scale (GCS) and Acute Physiologic and Chronic Health Evaluation (APACHE II). These findings suggest that the central venous plasma values of IL-8 at admission might be an early predictive marker in patients with severe TBI, comparative to standard clinical prognostic markers such as APACHE II and GCS. PMID- 17409680 TI - Successful treatment of advanced extranodal NK/T cell lymphoma with unrelated cord blood transplantation. AB - Nasal natural killer (NK)/T cell lymphoma is a rare entity of non-Hodgkin's lymphoma which mostly occurs in East Asian countries. The advanced disease above clinical stage III is often refractory to the radiation and chemotherapies, remission is transient even if achieved, and median survival is about 12 months. Thus the prognosis of advanced NK/T cell lymphoma is generally poor, however, the promising results of allogeneic hematopoietic stem cell transplantation for advanced NK/T cell lymphoma have been recently reported. In most of these cases, stem cell sources were human leukocyte antigen (HLA) matched donors and alternative sources were seldom used. We report here a case of a 36-year-old woman who was diagnosed as having an extranodal NK/T cell lymphoma, nasal type. The patient achieved a complete remission after 2 cycles of chemotherapy including Carboplatin, Etoposide, Ifosfamide, and Dexamethasone, but 3-months later relapsed during the search for HLA-matched unrelated donors. She received unrelated cord blood transplantation (CBT) in the second remission achieved by a regimen containing L-asparaginase. The conditioning regimen was 12 Gy of total body irradiation, high-dose cytarabin and cyclophosphamide. FK506 and methotrexate were used for graft-versus-host disease (GVHD) prophylaxis. GVHD involving the intestine and the oral mucosa was observed, but improved without additional immunosuppressive therapies. The patient remains in remission 33 months after CBT. Cord blood thus could be an appropriate stem cell source for patients with advanced NK/T lymphoma who have no HLA matched donors. PMID- 17409681 TI - Facial nerve conduction after sclerotherapy in children with facial lymphatic malformations: report of two cases. AB - Surgical excision is thought to be the standard treatment of choice for lymphatic malformations. However, when the lesions are limited to the face only, surgical scar and facial nerve injury may impair cosmetics and facial expression. Sclerotherapy, an injection of a sclerosing agent directly through the skin into a lesion, is an alternative method. By evaluating facial nerve conduction, we observed the long-term effect of facial lymphatic malformations after intralesional injection of OK-432 and correlated the findings with anatomic outcomes. One 12-year-old boy with a lesion over the right-side preauricular area adjacent to the main trunk of facial nerve and the other 5-year-old boy with a lesion in the left-sided cheek involving the buccinator muscle were enrolled. The follow-up data of more than one year, including clinical appearance, computed tomography (CT) scan and facial nerve evaluation were collected. The facial nerve conduction study was normal in both cases. Blink reflex in both children revealed normal results as well. Complete resolution was noted on outward appearance and CT scan. The neurophysiologic data were compatible with good anatomic and functional outcomes. Our report suggests that the inflammatory reaction of OK-432 did not interfere with adjacent facial nerve conduction. PMID- 17409683 TI - [Regulation of globotriaosylceramide (Gb3)-mediated signal transduction by rhamnose-binding lectin]. AB - Silurus asotus (catfish) egg lectin (SAL) has potent affinity to Gal alpha-linked carbohydrate chains of not only glycoproteins but also glycosphingolipids such as globotriaosylceramide (Gb3). SAL selectively bound to Gb3 localized in glycosphingolipid-enriched microdomain (GEM) of Gb3-expressing (Gb3(+)) Burkitt's lymphoma cells. Since treatment of Gb3(+) cells with SAL caused an increase in externalization of phosphatidylserine via activation of P-glycoprotein, and apoptotic volume decrease via activation of G-protein activated K(+) channel-1, SAL may function as an inducer of early apoptotic signal; however, neither caspase-8 and -3 activation nor DNA fragmentation was observed. We therefore investigated whether cell proliferation and viability were altered in SAL-treated Raji cells. SAL caused reduction of Raji cell proliferation without cytotoxicity. Although SAL did not induce apoptotic cell death to Gb3-expressing cells, it functionally behaved as a regulator of cell proliferation. SAL activated the suppression system of cell proliferation, such as down-regulation of c-myc and cdk4, and up-regulation of p21 and p27, inducing G1 arrest of the cell cycle, and consequently inhibited cell proliferation of Raji cells. Therefore, we conclude that SAL leads the cells to early apoptotic status but not late apoptotic (necrotic) status via binding to Gb3 existing in GEM, and that this binding is a prerequisite condition to induce cell cycle stop signal. PMID- 17409684 TI - [Regulation of neuronal cell function by glyco-signals]. AB - Gangliosides and proteoglycans with various sugar chains exist abundantly in the brain. They participate in intercellular recognition by revealing the sugar chains on the cell surface, and some of them show neurite-extension activity. Several recognition features that are mediated by the sugar chains are known such as saccharide-saccharide interaction and cell-surface sugar-chain receptor mediated recognition. Experiments on animals lacking the sugar-chain synthetic system with the technique of gene targeting suggest that phylogenetically "old" sugar chains such as chondroitin sulfate appear necessary for early development of the organism while relatively "new" sugar chains such as gangliosides, which appear with further development of the brain, are necessary for differentiation maturity processes. On the other hand, research using primary cultured neurons showed similar effects of the gangliosides and chondroitin sulfate on cell differentiation. It is possible that these sugar chains share the glyco-receptor mediated signal transduction system. PMID- 17409685 TI - [Regulation of integrin functions by N-glycans]. AB - Integrins are cell surface transmembrane glycoproteins that function as adhesion receptors in cell-ECM interactions and link matrix proteins to the cytoskeleton. Integrins play an important role in cytoskeleton organization and in the transduction of intracellular signals, regulating various processes such as proliferation, differentiation, apoptosis, and cell migration. Although integrin mediated adhesion is based on the binding of alpha and beta subunits to a defined peptide sequence, the strength of this binding is modulated by various factors including the status of glycosylation of integrin. Glycosylation reactions are catalyzed by the catalytic action of glycosyltransferases, such as N acetylglucosaminyltransferase III, V and alpha1,6 fucosyltransferase, etc., which catalyze the formation of glycosidic bonds. In this talk we will briefly overview the N-glycan structures of integrins, such as alpha3beta1 and alpha5beta1, and their related functions arising from recent studies, which provide insight into some long-standing questions concerning N-glycosylation functions. PMID- 17409686 TI - Insulin resistance as a membrane microdomain disorder. AB - Membrane microdomains (lipid rafts) are now recognized as critical for proper compartmentalization of insulin signaling, but their role in the pathogenesis of insulin resistance has not been investigated. Detergent-resistant membrane microdomains (DRMs), isolated in the low density fractions, are highly enriched in cholesterol, glycosphingolipids and various signaling molecules. TNFalpha induces insulin resistance in type 2 diabetes, but its mechanism of action is not fully understood. We have found a selective increase in the acidic glycosphingolipid ganglioside GM3 in 3T3-L1 adipocytes treated with TNFalpha, suggesting a specific function for GM3. We were able to extend these in vitro observations to living animals using obese Zucker fa/fa rats and ob/ob mice, in which the GM3 synthase mRNA levels in the white adipose tissues are significantly higher than in their lean controls. In the DRMs from TNFalpha-treated 3T3-L1 adipocytes, GM3 levels were doubled, compared to results in normal adipocytes. Additionally, insulin receptor (IR) accumulations in the DRMs were diminished, while caveolin and flotillin levels were unchanged. GM3 depletion was able to counteract the TNFalpha-induced inhibition of IR accumulation into DRMs. Together, these findings provide compelling evidence that in insulin resistance the insulin metabolic signaling defect can be attributed to a loss of IRs in the microdomains due to an accumulation of GM3. PMID- 17409688 TI - [Safe improvement of drug absorption by combinatorial use of sodium laurate with amino acids: cytoprotection by amino acids and its mechanisms]. AB - The development of combinatorial chemistry and high-throughput screening techniques has made it possible to generate many new drug candidates very rapidly, but it has also resulted in a number of poorly soluble and/or poorly absorbable candidates. A new trend in drug development based on pharmacogenomics or the development of molecular-targeted drugs is also spurring the tendency, and it does not necessarily lead to good output in terms of the development of new drugs. It is attractive to improve membrane permeability as well as solubility by using adjuvants, because this method could be applicable for various drugs. However, the practical use of absorption-enhancing adjuvants has been limited because of the potential local toxicity. Therefore suppressing the potential local toxicity would lead to the successful development of safe preparations with improved absorption using adjuvants. Our biochemical and histopathologic studies showed that several amino acids such as taurine and L-glutamine had cytoprotective activity, and it has been found that the combinatorial use of sodium laurate (C12) with these amino acids could maintain the absorption enhancing ability of C12. A suppository preparation containing C12 and taurine remarkably improved the rectal absorption of rebamipide, classified as BCS class IV, and the preparation was safe to the rectal mucosa. For the mechanisms of cytoprotective action by these amino acids, it has been found that they suppress the intracellular calcium level, induce the expression of heat-shock protein 70, and inhibit the release of histamine and apoptosis. PMID- 17409689 TI - [Barriology-based strategy for drug absorption]. AB - Passing of drugs across epithelial cell sheets and endothelial cell sheets is an obligatory step in the absorption of a drug. The passing routes of drugs are classified into transcellular and paracellular pathways. The transcellular route has been widely investigated and is used in clinical therapy. In contrast, drug delivery using the paracellular route has never been fully developed. Sodium caprate is the only absorption-enhancer of drugs that uses the paracellular route. Tight junctions (TJs) exist between adjacent cells in epithelial and endothelial cell sheets, and they play a role in sealing the cell sheets. Therefore, we must modulate the TJ barrier for drug delivery using paracellular route. In this review, we describe barriology, including very recent topics, and overview absorption-enhancers from the perspective of barriology. PMID- 17409690 TI - [Hydrolysis by carboxylesterase and disposition of prodrug with ester moiety]. AB - Prodrug is a useful approach for improving the bioavailability of therapeutic agents through increased passive transport. Carboxylesterases (CESs, EC.3.1.1.1.) that show ubiquitous expression profiles play an important role in the biotransformation of ester-containing prodrugs into their therapeutically active forms in the body. High levels of CESs are found in the liver, small intestine and lungs where prodrugs are firstly hydrolyzed before entering the systemic circulation. Rat intestine single-pass perfusion experiments have shown that CES is involved in the intestinal first-pass hydrolysis. Extensive pulmonary first pass hydrolysis has been observed in accordance to the substrate specificity of CES1 isozyme. Hydrolysis in the human liver and lungs is mainly catalyzed by hCE1 (a human CES1 family isozyme), whereas that in the small intestine is predominantly mediated by hCE2 (a human CES2 family isozyme). hCE2 preferentially hydrolyzes substrates with a small acyl moiety such as CPT-11, due to conformational steric hindrance in its active site. In contrast, hCE1 is able to hydrolyze a variety of substrates due to spacious and flexible substrate binding region in its active site. In addition, hCE1 has been found to catalyze transesterification. Caco-2 cells mainly expresses CES1 isozyme but not CES2 isozyme. Because of the differences in substrate specificity between CES1 and CES2 enzymes, Caco-2 cell monolayer is not suitable for predicting intestinal absorption of prodrugs. These findings indicate that identification of substrate specificity of CES isozymes and development of an in vitro experimental method are essential to support rational design of prodrug. PMID- 17409691 TI - [Study on the colon specific delivery of prednisolone using chitosan capsules]. AB - In this study, we examined the effectiveness of chitosan capsules for the colon specific delivery of prednisolone in rats. We also evaluated the effectiveness and side effects of prednisolone using chitosan capsules compared with the conventional dosage form (gelatin capsules). We found a significant increase in the concentration of prednisolone in the large intestinal mucosa when prednisolone was administered orally using chitosan capsules, as compared with the case using gelatin capsules. On the other hand, the plasma concentrations of prednisolone after oral administration using chitosan capsules were much lower than those in the case of gelatin capsules. We also assessed the effectiveness of prednisolone for the healing of trinitrobenzene sulfonic acid-induced colitis by measuring myeloperoxidase (MPO) activity and colon wet weight/body weight (C/B) ratio. MPO activities and C/B ratios were significantly reduced when prednisolone was administered orally using chitosan capsules, in comparison with the case of gelatin capsules. Moreover, the weight of the thymus, which is an index of the side effects of prednisolone, markedly decreased after oral administration of prednisolone using gelatin capsules, whereas its weight did not change as much when prednisolone was administered orally using chitosan capsules. These findings indicate that chitosan capsules might be useful for the colon-specific delivery of prednisolone and its enhanced effectiveness for the healing of colitis in rats. Moreover, chitosan capsules might be also effective in reducing the side effects of prednisolone due to its decreased intestinal transfer to the systemic circulation. PMID- 17409692 TI - [Control of pulmonary absorption of drugs by various pharmaceutical excipients]. AB - In general, drugs are well absorbed from the lung, and the pulmonary absorption of therapeutic protein and peptide drugs, which are poorly absorbed from the gastrointestinal tract, was observed. However, locally acting drugs including antiasthmatic agents, bronchodilators, and expectorants should be localized for a long period in the lung tissues. In this study, the effects of various viscous vehicles on the absorption of theophylline and fluticasone propionate after intrapulmonary administration were examined in rats. Carrageenans were effective in regulating the absorption rate of these drugs. On the other hand, the bioavailability of therapeutic protein and peptide drugs with relatively high molecular weights from the pulmonary route is still poor when compared with the parenteral route. Therefore we examined the effects of chitosan and chitosan oligomers on the pulmonary absorption of interferon-alpha and salmon calcitonin in rats. Chitosan oligomers were effective in improving the pulmonary absorption of these drugs, and chitosan hexamer appeared to be markedly more effective than other oligomers. Furthermore, the present study indicated that chitosan oligomers did not cause any membrane damage to rat pulmonary tissues. In conclusion, it is suggested that various pharmaceutical excipients achieved the sustained pulmonary absorption of locally acting drugs and the improved pulmonary bioavailability for therapeutic protein and peptide drugs. PMID- 17409693 TI - [Local and systemic delivery of high-molecular weight drugs by powder inhalation]. AB - The pulmonary route has recently attracted attention as a noninvasive administration route for peptide and protein drugs, and an insulin powder for inhalation was approved by authorities in Europe and the USA. The present study examined usefulness of insulin and gene powders for systemic and local inhalation therapy. We prepared several dry insulin powders by spray drying to examine the effect of additives on insulin absorption. Citric acid appears to be a safe and potent absorption enhancer for insulin in dry powder. However, in the powder with citric acid (MIC0.2 SD) insulin was unstable compared with the other powders examined. To improve insulin stability, a combination of insulin powder and citric acid powder was prepared (MIC Mix). MIC Mix showed hypoglycemic activity comparable to MIC0.2 SD while the insulin stability was much better than that of MIC SD. Next, dry insulin powders with mannitol were prepared with supercritical carbon dioxide (SCF); the powder thus prepared reduced blood glucose level rapidly and was more effective than that prepared by spray drying. Chitosan-pDNA complex powders as a pulmonary gene delivery system were also prepared with SCF and their in vivo activity was evaluated. The addition of chitosan suppressed the degradation of pCMV-Luc during preparation and increased the storage stability. The luciferase activity in mouse lung was evaluated after pulmonary administration of the powders. The chitosan-pDNA powder with an N/P ratio=5 increased the luciferase activity to 27 times that of the pCMV-Luc solution. These results suggest that gene powder with chitosan is a useful pulmonary gene delivery system. PMID- 17409694 TI - [Recent trend of transdermal drug delivery system development]. AB - The potential of transdermal drug delivery systems has been demonstrated in recent years with the approval of several medicines for use by patients who are unable to use conventional dosage routes, like oral administration or injection. To enhance the TDDS (Transdermal Drug Delivery System) potential to include other drug candidates, many researchers have been exploring enhancement approaches to increase the permeability of various drugs through the skin. Recently, physical enhancement systems are being reported as having big potential by many researchers. In particular, iontophoresis is a very attractive way of delivering ionized drugs by the application of an electric field to the skin. This has been marketed with some topical and systemic drugs (lidocaine and fentanyl). Sonophoresis is also an attractive method to deliver a drug through the skin using ultrasound. Besides these technologies, various physical approaches are under study. Such technologies can be expected to deliver not only small MW compounds but also macromolecules like peptides. In this article, after looking back through the history of TDDS development, I would like to summarize with new physical and chemical approaches and outline of the new trend of TDDS development with those enhancement system. PMID- 17409695 TI - Stress Responses via Metallothioneins: Roles in the Biological Defense Mechanism. PMID- 17409696 TI - [Heavy metal responses of the human metallothionein isoform genes]. AB - Metallothioneins (MTs) are proteins known to be involved in defense mechanisms against heavy metals and reactive oxygen species. In human, more than ten MT isoform genes have been identified, in contrast to much fewer isoforms in other mammalian species. The increased number of isoforms in human may have some biological significance; for example, isoforms may have been functionally differentiated to deal with various environmental factors in the evolutional process. However, we know little about the functions of the individual MT isoforms. To clarify functional differences between human MT isoforms, we developed a method to determine individual isoform mRNA levels using real-time polymerase chain reaction (PCR), and studied responses of the isoform genes against heavy metals (Zn, Cd, Cu) and As in HeLa cells. These metals induced all MT isoforms except for MT-1A by Cu, though their induced levels were different. Furthermore, these metals preferentially induced isoforms MT-2A and MT-1X suggesting that these isoforms may be important in protecting from their cytotoxicity. PMID- 17409697 TI - [Mechanism of metallothionein gene activation mediated by heavy-metal dependent transcription factor MTF-1]. AB - Transcriptional activation of metallothionein (MT) genes by heavy metals is a valuable system for understanding the functions of MT as well as the cellular response against heavy metals. Although it is now known that heavy metal signals culminating in MT induction converge upon a transcription factor MTF-1, the mechanism underlying the MTF-1 response to heavy metals has not been elucidated. To address this issue, we investigated various aspects of the in vivo response of MTF-1 against heavy metals. Chromatin immunoprecipitation assay showed that heavy metal-dependent DNA binding of MTF-1 is the critical step in vivo. MTF-1 is primarily localized in the nucleus so that heavy metal-dependent nuclear translocation demonstrated by other groups does not seem to be universal and hence may not be critical for activation of MTF-1. In the six Zn finger motifs, the hallmark of MTF-1, the third and the fourth fingers are essential for the nuclear localization of MTF-1. Furthermore, all fingers except the last are important for transcriptional activation function of MTF-1, suggesting their key role for MTF-1 function. Also, a cysteine cluster structure located in the C terminal region of MTF-1 is critical for transactivating function of MTF-1. These results suggest a central role of the Zn-finger domain and intramolecular cooperation through a structural change of MTF-1 for its response to heavy metal challenge. PMID- 17409698 TI - [Cytokine-induced metallothionein expression and modulation of cytokine expression by metallothionein]. AB - A multifunctional protein metallothionein (MT) is induced by various chemicals and cytokines. We have found novel functions of MT as follows: 1) Cytokine expression such as IL-1alpha, IL-6, and TNFalpha responding to lipopolysaccharide is reduced in MT-deficient macrophages compared with in wild-type cells. 2) Nitric oxide production responding to TNFalpha and LPS is reduced in MT-deficient macrophages compared with in wild-type cells. 3) M-CSF expression responding to zinc is reduced in MT-deficient fibroblasts compared with in wild-type cells, and increased in MT-overexpressed fibroblasts compared with in control cells. 4) LIF, a STAT3 activating cytokine, protects the heart from ischemia/reperfusion injury. Transgenic mice overexpressing STAT3 have tolerance to ischemia/reperfusion induced damage, whereas MT-null mutation cancels the myocardial protection. In this review, we discuss the relation of MT and stress responses from the point of view of cytokine-induced expression of MT and modulation of cytokine expression by MT. PMID- 17409699 TI - [Physiological significance of metallothionein in oxidative stress]. AB - Metallothionein (MT), a ubiquitous family of low-molecular weight metal-binding proteins, comprises 30% cysteine residues. Although all of the thiol residues in MT are bound to metals, it still remains active to reactive oxygen species. Each cysteine residue in MT is more effective at protecting DNA from hydroxyl radical attack than the glutathione cysteine in vitro. Prooxidative agents such as paraquat and carbon tetrachloride induce MT synthesis mediated by some responsive elements. MT demonstrates strong antioxidant properties, yet the physiological relevance of its antioxidant action is not clear. An injection of ferric nitrilotriacetate (Fe-NTA), which produces reactive oxygen species, caused transcriptional induction of MT synthesis in the liver and kidney. Pretreatment of mice with Zn attenuated nephrotoxicity induced by Fe-NTA. After a Fe-NTA injection, a loss of Cd-binding properties of preinduced MT was observed only in kidneys of Zn-pretreated mice but not in liver. MT-enriched hepatocytes are resistant to Fe-NTA toxicity, oxidative DNA, and cell damage during conditions of glutathione depletion. In glutathione-depleted cells, but not in non-treated cells, Cd-binding properties of cellular MT decreased with increasing concentration of Fe-NTA. Moreover, Cd released from MT after an injection of Fe NTA induced new MT protein again. Thus MT may act as a secondary antioxidant in cellular protection system against oxidative stress. PMID- 17409700 TI - [Endoplasmic reticulum stress and metallothionein]. AB - Much attention has been paid to lifestyle-related diseases including type 2 diabetes mellitus, cardiovascular disease, hypertension, and hyperlipidemia because the incidence rates of these diseases are increasing in developed countries. Elucidation of factors contributing to the development of obesity and insulin resistance is needed. Metallothionein (MT), a ubiquitous metal-binding protein, is induced not only by heavy metals but also by various kinds of stresses. Endoplasmic reticulum (ER) stress is caused by accumulation of misfolded proteins in ER. Recently, increased ER stress by obesity and impairment of insulin action by ER stress have been reported. Exposure to ER stress increased induction of MT synthesis, and an enhanced response to ER stress evaluated as expression of Bip/GRP78mRNA was observed in the liver of MT-null mice, suggesting that MT attenuates expression of ER stress. MT may prevent ER stress and thereby modulate the development of obesity and insulin resistance. A possible role of metallothionein in response reaction for ER stress is discussed. PMID- 17409701 TI - [Analysis of toxicity using metallothionein knockout mice]. AB - Two research groups produced metallothionein (MT)-I/II knockout mice with null mutation of MT-I and MT-II genes. In 1993, Choo et al. produced MT-I/II knockout mice with a mixed genetic background of 129 Ola and C57BL/6 strains. Palmiter et al. also produced MT-I/II knockout mice with a genetic background of 129/Sv strain in 1994. Subsequently, MT-I/II knockout mice have been used to clarify the biological function and physiological role of MT by many research groups. We were also provided MT-I/II knockout mice from Dr. Choo (Australia). F1 hybrid mice were mated with C57BL/6, and their offspring were back-crossed to C57BL/6 for ten generations. MT-I/II knockout (MT(-/-)) mice and wild-type (MT(+/+)) mice were obtained by mating of those heterozygous (MT(+/-)) mice. We have been investigating the susceptibility of MT-I/II knockout mice to toxicity of harmful factors and some diseases. Our present studies found that MT-I/II knockout mice have an increased sensitivity to harmful metals such as cadmium, mercury, and arsenic, oxidative stress, chemical carcinogenesis and neurodegenerative diseases. These results clearly indicate that MT plays an important role in defense of these toxicities. In this review, we present our findings and summarize recent reports with MT-I/II knockout mice concerning the role of MT as a biological protective factor. PMID- 17409703 TI - [Study for an allergic inflammation model using human lungs and its pharmacological application]. AB - The complement system, which plays an important role in inmate immunity, is considered to be important in the pathophysiology of allergic asthma. A patient with allergic asthma shows the reversible characteristic system of bronchoconstriction, increased mucus secretion, and complicated airway inflammation. Various cytokines secreted from Th2 cells contribute to the system. Cysteinyl-leukotrienes (CysLTs) are also considered to be one of the important mediators involved in asthmatic pathophysiology. However, the effects of a drug on humans may not be the same as those on animals due to species differences in complement-related molecules. In this series of experiments, we tried to establish a model in which the effects of a drug on the production of CysLTs from human lung preparations were evaluated following an anaphylactic reaction. CysLT production increased when the passively sensitized lung tissues were stimulated with anti-IgE antibody. The coaddition of anaphylatoxin, C5a, with the anti-IgE antibody potentiated CysLT production. The response to C3a was weaker when compared with that to C5a. In addition, increased production of CysLTs by adding serum at a specific ratio was dose dependently inhibited by nonpeptide C5a receptor antagonist, W-54011, or a novel complementary peptide inhibitor of C5a, acetyl peptide A. From these results, it is suggested that C5a potentiates cysLT production from human lung tissues and contributes to allergic inflammation like asthma, and thus acetylated peptide A and W-54011 are useful for suppressing allergic inflammation in the lungs. PMID- 17409704 TI - [Mechanisms underlying enhanced vasodilator responses to various vasodilator agents following endothelium removal in rat mesenteric resistance arteries]. AB - We reported that vasodilator responses to various vasodilator agents were augmented by endothelium removal. To explain this mechanism, we hypothesized that endothelium removal eliminates the release of endothelium-derived contracting factor EDCF, which counteracts the vasodilation. However, the underlying mechanism is unknown. Therefore the present study investigated the second messenger system further to investigate the mechanisms underlying enhanced vasodilator response after endothelium removal in rat mesenteric resistance arteries. Mesenteric vascular beds isolated from Wistar rats were perfused and perfusion pressure was measured. The vascular endothelium was removed by 30-s perfusion of sodium deoxycholate. Vasodilator responses to sodium nitroprusside (SNP) perfusion were markedly augmented and prolonged by endothelium removal. In preparations with intact endothelium and active tone, 5-min perfusion of sodium azide (non-specific guanylate cyclase (GC) activator), ANP (membrane-linked GC activator), and 8-Br-cGMP (cGMP analogue) caused a concentration-dependent vasodilation that was markedly augmented by endothelium removal. However, vasodilation induced by YC-1 and BAY41-2272 (selective soluble GC activator) was not augmented by endothelium removal. When methylene blue (soluble GC inhibitor) was present in the medium, SNP caused a concentration-dependent vasodilation in the preparation with intact endothelium, which was less augmented by endothelium removal compared with control (preparation without methylene blue). These findings suggest that endothelium removal affects intracellular cGMP-mediated signal transduction system in vascular smooth muscle cells. PMID- 17409705 TI - [Strategy to develop a new drug for treatment-resistant depression--role of electroconvulsive stimuli and BDNF]. AB - In recent years, depression studies have focused on morphological changes associated with depression. Brain-derived neurotrophic factor (BDNF) is a neurotrophic factor that plays an important role in the morphological changes associated with depression and the mechanisms of antidepressants. On the other hand, hyperfunction of the hypothalamic-pituitary-adrenal axis has been link to pathophysiology of depression. In our previous studies, ACTH-treated rats served as a valuable animal model of tricyclic antidepressant-resistant depressive conditions. However, few neuroanatomic studies have been done. In the present study, we investigated mechanisms underling ACTH-treated rat serving an imipramine treatment-resistant depression model using c-Fos as a marker. The c Fos immunohistochemical study indicated that the medial prefrontal cortex is an action site of imipramine in ACTH-treated rats. Electroconvulsive therapy is considered an effective treatment for treatment-resistant depression. However, the mechanisms causing treatment-resistant depressive conditions are unknown. We investigated the effect of repeated electrical convulsive shock (ECS)-treatment using the forced swim test, a screening method for antidepressant-like activity, and hippocampal BDNF protein levels in ACTH-treated rats. Findings showed that repeated ECS treatment decreased the immobility time during forced swim test. Furthermore, the ECS treatment also markedly increased the hippocampal BDNF levels in the rat tricyclic antidepressant-resistant depression model. In addition, the repeated ECS treatment showed long-lasting effects on forced swim test and increased of hippocampal BDNF levels in normal rats. These findings suggest that BDNF plays a key role in the antidepressant-like effect of ECS and that increased BDNF may be involved in promoting the long-lasting effect. PMID- 17409706 TI - [Brain embolism-induced injury of vascular endothelial cells and a novel vasoprotective drug]. AB - Microsphere embolism-induced up-regulation of endothelial nitric oxide synthase (eNOS) in endothelial cells of brain microvessels was found after brain ischemia. The eNOS induction preceded disruption of the blood-brain barrier following ischemia. In vascular endothelial cells, microsphere embolism-induced eNOS expression was associated with protein tyrosine nitration, which is a marker of generation of peroxynitrite. To determine whether eNOS expression and protein tyrosine nitration in vascular endothelial cells mediates the blood-brain barrier disruption in the microsphere embolism brain, we tested the effect of a novel calmodulin-dependent NOS inhibitor, DY-9760e, which inhibits eNOS activity and in turn protein tyrosine nitration. Concomitant with inhibition of protein tyrosine nitration in vascular endothelial cells, DY-9760e significantly inhibited BBB disruption as assessed by Evans blue excretion. DY-9760e also inhibited cleavage of poly(ADP-ribose) polymerase as a marker of the apoptotic pathway in vascular endothelial cells. Taken together with previous evidence in which DY-9760e inhibited brain edema, microsphere embolism-induced eNOS expression in vascular endothelial cells likely mediates BBB disruption and in turn brain edema. PMID- 17409707 TI - [Metabolism of non-steroidal anti-inflammatory drugs by peroxidase: implication for gastrointestinal mucosal lesions]. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat inflammatory diseases including rheumatoid arthritis and gout. The anti inflammatory action of NSAIDs is due to the inhibition of prostaglandin synthesis by preventing cyclooxygenase (COX) activity of prostaglandin H synthase (PGS). However, administration of NSAIDs causes gastrointestinal mucosal lesions and a decrease of granulocytes as side effects. PGS catalyzes two distinct enzyme reactions: (1) bis-dioxygenation of arachidonic acid catalyzed by COX activity of PGS to form PGG(2); and (2) reduction of the hydroperoxide group in PGG(2) by PGS hydroperoxidase. Most NSAID are oxidized by peroxidases to produce NSAID radicals that damage biological components such as lipids and enzymes. Indomethacin, phenylbutazone, and piroxicam are more toxic under aerobic conditions than anaerobic conditions during the interaction with peroxidase. We discuss the contribution of peroxidases in the formation of gastrointestinal mucosal lesions induced by NSAIDs. PMID- 17409708 TI - [Visualization and evaluation of the promoter activities of genes for stress inducible proteins in response to environmental pollutants]. AB - We examined the systematic assay of the reporter gene for the assessment of heavy metals and organic chemical pollutants using the reporter plasmids carrying stress-responsive elements fused to the green fluorescence protein (GFP) gene as follows: metallothionein (MTIIA), heme oxigenase-1 (HO-1), quinone reductase (ARE), and c-fos genes. The treatment of COS7 cells in which the c-fos gene promoter-, ARE-, or HO-1 enhancer-fused GFP with a low concentration of NaAsO(2) was introduced led to the detection of the fluorescent cells, and an agrichemical paraquat enhanced the fluorescence of ARE or HO-1 enhancer-transfected cells. The cells in which the plasmid carrying the MT-IIA gene promoter (the -765 bp upstream from the transcription initiation site) was introduced highly expressed GFP on treatment with CdCl(2), ZnSO(4), or CuCl(2). The plasmid carrying seven metal-responsive elements of the MT-IIA gene increased the response of the fluorescence intensity to these heavy metals. These results indicated that the use of the gene promoters and enhancers of the stress-responsive genes fused to GFP contributes to the visualization of pollutant-responsive mammalian cells and can be applied to biomonitoring of environmental pollution. PMID- 17409709 TI - [Comparison of the visual analog scale method and 5-point evaluation in student self-assessment of comprehension and acquisition in a model core curriculum for practical training]. AB - In examining assessment methods used for evaluating training, there have so far been no studies reporting any differences between the visual analogue scale (VAS) evaluation method, based on a rating scale, and evaluation methods based on an ordinal scale. Here we report the findings of an examination into differences and discrepancies between the results of the VAS method and a 5-point evaluation. Following the end of their training period, seven trainees carried out a self evaluation regarding their level of understanding and performance using the 5 point evaluation and VAS methods. We then compared the average results of both assessment methods and examined the correlation between the two sets of figures. We found no differences between the 5-point evaluation method and VAS method in evaluating training for dispensing drugs, administering injections, pharmacy preparation, and medication management and instruction. There was also a significant correlation between average values for the 5-point evaluation and VAS method in evaluating training for dispensing drugs, administering injections, pharmacy preparation, and medication management and instruction. This led us to the conclusion that both the 5-point evaluation method and VAS method give similar results and outcomes in assessing the results of practical training. PMID- 17409710 TI - [Inhibition of hyphal growth of the fungus Alternaria alternata by chlorine dioxide gas at very low concentrations]. AB - The efficacy of chlorine dioxide (ClO(2)) gas at very low concentrations for hyphal growth of Alternaria alternata related to fungal allergy was evaluated using a fungus detector. The fungus detector is a plastic sheet with a drop of spore-suspending medium, and it makes possible clear observations of hyphal growth with a light microscope. ClO(2) gas (average 0.075 ppm, 0.21 microg/l) inhibited hyphal growth of the fungus, but not germination of fungal spores. The hyphal length was more than 1780 mum under air conditions (control) and 49+/-17 microm under ClO(2) gas conditions for 72 h. According to the international chemical safety card, threshold limit values for ClO(2) gas are 0.1 ppm as an 8-h time-weight average and 0.3 ppm as a 15 min short-term exposure limit. From these data, we propose that treatment with ClO(2) gas at very low concentrations in space is a useful tool for the growth inhibition of fungi in the fields of food, medicine, etc. without adverse effects. PMID- 17409711 TI - Comparative analyses of song complexity and song-control nuclei in fourteen oscine species. AB - Most studies on the relationship between measures of song behavior and the sizes of song control nuclei have focused on one or two oscine species, and often show inconsistent results. To address this issue, we first measured four variables for song complexity, i.e., song repertoire size, syllable repertoire size, the mean number of syllables per phrase (MNS) and the number of syllables in the longest phrase (NSLP), and the sizes of three song control nuclei, i.e., HVC, RA (the robust nucleus of the arcopallium), and Area X in 14 oscine species from eight families. To tackle the problem of statistical non-independence that probably existed among the closely related species, we reconstructed the phylogeny of the species studied using mitochondrial cytochrome b DNA sequences from GenBank. By using the methods adopted in most previous reports, we tested the relationship between song complexity and the sizes of the song control nuclei. We found that: 1) the absolute sizes of RA and Area X, but not of HVC, were positively correlated to the three measures of song complexity, but that only the residual size of RA in regard to telencephalon size was significantly correlated to the song measures; 2) independent contrasts analysis showed RA and Area X to be significantly associated with NSLP. Our results indicated that the relationship between song behavior and its neural structures varied among song nuclei, suggesting that each song control nucleus may play a different role in song behavior. PMID- 17409712 TI - The conservation-refugium value of small and disturbed Brazilian Atlantic forest fragments for the endemic ovoviviparous cockroach Monastria biguttata (Insecta: Dictyoptera, Blaberidae, Blaberinae). AB - The Brazilian Atlantic forest is a biodiversity hotspot and harbors many endemic species showing peculiar and unique traits. However, it has been reduced to less than 8% of its original surface and is distributed in scattered fragments, the great majority of which are smaller than 20 hectares and very disturbed, making it worth asking about their value for conservation. In this paper we assess the refugium value of small fragments to the conservation of one of the endemics of the Atlantic forest, the ovoviviparous cockroach Monastria biguttata. Our results showed that this species was ubiquitous in large and small forest fragments, but never present in plantations or pastures. The population age structure and sex ratio were balanced in every fragment, and total population size in the smallest fragments was at least several hundreds of individuals. Colony size, sex ratio, age structure, and density per piece of dead tree trunk indicated that populations from small fragments were not unbalanced or at risk of extinction. According to the analysis of resource availability, small fragments can provide suitable habitat for this species. In this situation, even very small forest fragments have a high refugium value for some endemic insect species. Considering their number in the landscape, these fragments should be considered with more attention in strategies of biodiversity conservation. PMID- 17409713 TI - Drosophila CTLA-2-like protein (D/CTLA-2) inhibits cysteine proteinase 1 (CP1), a cathepsin L-like enzyme. AB - In this study, we present a propeptide-like cysteine proteinase inhibitor, Drosophila CTLA-2-like protein (D/CTLA-2), a CG10460 (crammer) gene product, with an amino acid sequence significantly similar to the proregion of Drosophila cysteine proteinase 1 (CP1). Recombinant D/CTLA-2, expressed in E. coli, strongly inhibited Bombyx cysteine proteinase (BCP) with a Ki value of 4.7 nM. It also inhibited cathepsins L and H with Ki values of 3.9 (human liver) and 0.43 (rabbit liver) nM, and 7.8 nM (human liver), respectively. Recombinant D/CTLA-2 exhibited low but significant inhibitory activities to cathepsin B with Ki values of 15 nM (human liver) and 110 nM (rat liver), but hardly inhibited papain. We attempted to purify cysteine proteinases inhibited by D/CTLA-2 from total bodies of adult Drosophila. Recombinant D/CTLA-2 significantly inhibited CP1 with a Ki value of 12 nM, indicating that CP1, a cognate enzyme of D/CTLA-2, is a target enzyme of the inhibitor in Drosophila cells. These results indicate that D/CTLA-2 is a selective inhibitor of cathepsin L-like cysteine proteinases similar to other propeptide-like cysteine proteinase inhibitors such as Bombyx cysteine proteinase inhibitors (BCPI) and cytotoxic T-lymphocyte antigen-2 (CTLA-2). D/CTLA-2 was expressed over the whole life cycle of Drosophila. Strong expression was observed in the garland cells and prothoracic gland in the late stages of embryonic development. These results suggest that D/CTLA-2, implicated in intra- and extra cellular digestive processes, functions in these tissues by suppressing uncontrolled enzymatic activities of CP1. PMID- 17409714 TI - The Dugesia ryukyuensis database as a molecular resource for studying switching of the reproductive system. AB - The planarian Dugesia ryukyuensis reproduces both asexually and sexually, and can switch from one mode of reproduction to the other. We recently developed a method for experimentally switching reproduction of the planarian from the asexual to the sexual mode. We constructed a cDNA library from sexualized D. ryukyuensis and sequenced and analyzed 8,988 expressed sequence tags (ESTs). The ESTs were analyzed and grouped into 3,077 non-redundant sequences, leaving 1,929 singletons that formed the basis of unigene sets. Fifty-six percent of the cDNAs analyzed shared similarity (E-value<1E -20) with sequences deposited in NCBI. Highly redundant sequences encoded granulin and actin, which are expressed in the whole body, and other redundant sequences encoded a Vasa-like protein, which is known to be a component of germ-line cells and is expressed in the ovary, and Y protein, which is expressed in the testis. The sexualized planarian expressed sequence tag database (http://planaria.bio.keio.ac.jp/planaria/) is an open access, online resource providing access to sequence, classification, clustering, and annotation data. This database should constitute a powerful tool for analyzing sexualization in planarians. PMID- 17409715 TI - The role of cuticular hydrocarbons in mating and conspecific recognition in the closely related longicorn beetles Pidonia grallatrix and P. takechii. AB - The role of cuticular chemicals in mating behavior and their chemical components were studied in two sympatric flower-visiting longicorn beetles, Pidonia grallatrix and P. takechii. Mating experiments revealed that female cuticular chemicals elicit male mating behavior and that males can discriminate between conspecific and heterospecific females on the basis of contact chemicals. GC-MS analyses of whole-body extracts in the two species and both sexes determined that extracts contained a series of hydrocarbons including n-alkanes, n-alkenes, and methylalkanes. The relative abundance of some hydrocarbons differed between species and sexes, and canonical discriminant analysis showed discrimination of species and sex could be made unambiguously with several compounds. These results imply that the difference in cuticular hydrocarbons facilitates the premating isolation of sympatric Pidonia species. PMID- 17409716 TI - Substratum and morphometric relationships in the bryozoan genus Odontoporella, with a description of a new paguridean-symbiont species from New Zealand. AB - Members of the bryozoan family Hippoporidridae have frequently been found encrusting gastropod shells inhabited by hermit crabs, with which they appear to enter into a symbiotic relationship-shells occupied by hermits may in some species have a tubular extension of the encrusting bryozoan from the shell opening, induced by the presence of the crab. Such colony growth is characteristic of some species of Hippoporidra Canu and Bassler and Odontoporella Hejjas. The type species of Odontoporella, O. adpressa (Busk), has been attributed a nominal distribution from Chiloe Island, Chile (the type locality), to the Falkland Islands, New Zealand, and New Caledonia. In New Zealand, colonies are relatively easily obtainable from some localities, so a study was undertaken to ascertain substratum and morphometric relationships across the range of distribution of the species, using museum specimens and, where possible, fresh material. It became clear that the New Zealand population constitutes a separate species, here named O. bishopi n. sp., in which the orifice is proportionately larger than in O. adpressa. In contrast to O. adpressa, which settles on a range of substrata, O. bishopi preferentially settles on gastropod shells occupied by hermit crabs (mostly Paguristes setosus (H. Milne Edwards)) and shows sexual dimorphism at the level of the polypide. Male polypides not only have modified lophophores but also reduced guts. PMID- 17409717 TI - Phylogeny and phylogeography of the genus Geothelphusa (Crustacea: Decapoda, Brachyura, Potamidae) in southwestern Taiwan based on two mitochondrial genes. AB - Eleven species of Geothelphusa have been reported from southwestern Taiwan (Tainan, Kaohsiung and the northern part of Pingtung counties): G. albogilva Shy, Ng, and Yu, 1994; G. ancylophallus Shy, Ng, and Yu, 1994; G. caesia Shy, Ng, and Yu, 1994; G. lili Chen, Cheng, and Shy, 2005; G. nanhsi Shy, Ng, and Yu, 1994; G. neipu Chen, Cheng, and Shy, 1998; G. olea Shy, Ng, and Yu, 1994; G. pingtung Tan and Liu, 1998; G. shernshan Chen, Cheng, and Shy, 2005; G. tsayae Shy, Ng, and Yu, 1994 and G. wutai Shy, Ng, and Yu, 1994. Comparisons of DNA sequences encoding parts of the mitochondrial large subunit (16S) rRNA and cytochrome oxidase subunit I (COI) genes revealed three major clades, of which one is the species G. ancylophallus, and the other two are species groups here referred to as the G. olea and G. pingtung clades. Geothelphusa ancylophallus is geographically restricted and adapted to an ecologically challenging habitat with an unstable water supply and uneven topology. The G. olea clade (G. olea, G. caesia, G. nanhsi, G. tsayae, and G. wutai) is widely distributed throughout central-western and southwestern Taiwan. The G. pingtung clade (G. pingtung, G. neipu and G. shernshan) is confined to southwestern Taiwan between the previously defined southernmost clades of G. tawu, G. albogilva, and G. ferruginea, and the G. olea clade to the north. It includes an isolated population on distant Chaishan Mountain near Taiwan Strait, which probably dispersed from the peripheral hills of the Central Range during the early Pleistocene. The available genetic evidence indicates that the differential coloration observed in members of the G. olea and G. pingtung clades is not reflected in mtDNA, appears to be dependent on environmental conditions, food, etc., and has little value as a taxonomic character. Possible geological events and climatic factors responsible for the historic isolation of the different freshwater crab clades in southwestern Taiwan are discussed in detail. PMID- 17409718 TI - Lack of circadian regulation of melatonin rhythms in the sockeye salmon (Oncorhynchus nerka) in vivo and in vitro. AB - Melatonin profiles were determined in the plasma in vivo and in the pineal organ in vitro of the sockeye salmon (Oncorhynchus nerka) under various light conditions to test whether they are under circadian regulation. When serial blood samples were taken at 4-h intervals for 3 days via a cannula inserted into the dorsal aorta, plasma melatonin exhibited significant fluctuation under a light dark cycle, with higher levels during the dark phase than during the light phase. No rhythmic fluctuations persisted under either constant dark or constant light, with constant low and high levels, respectively. Melatonin release from the pineal organ in flow-through culture exhibited a similar pattern in response to the change in light conditions, with high and low release associated with the dark and light phases, respectively. These results indicate that melatonin production in the sockeye salmon is driven by light and darkness but lacks circadian regulation. PMID- 17409719 TI - Phylogenetic relationships of the Chinese brown frogs (genus Rana) inferred from partial mitochondrial 12S and 16S rRNA gene sequences. AB - Based on partial sequences of the 12S and 16S ribosomal RNA genes, we estimated phylogenetic relationships among brown frogs of the Rana temporaria group from China. From the phylogenetic trees obtained, we propose to include Rana zhengi in the brown frogs. Monophyly of the brown frogs was not unambiguously supported, but four well-supported clades (A, B, C, and D) always emerged, although relationships among them remained unresolved. Clade A contained brown frogs with 24 chromosomes and was split into two distinct subclades (Subclade A-1: R. chensinensis and R. huanrenensis; Subclade A-2: R. dybowskii). Polytomous relationships among populations of R. chensinensis and R. huanrenensis suggested the necessity of further taxonomic assessment. Rana kunyuensis proved to be the sister group to R. amurensis, and these two species formed Clade B. Clade C was composed of R. omeimontis and R. chaochiaoensis, and Clade D included R. sauteri, which has been placed in other ranid genera. These relationships did not change after adding published data, and monophyly of Subclade A-1, A-2, and other East Asian brown frogs with 24 chromosomes (R. pirica and R. ornativentris) was ascertained, though their relationships were unresolved. Clade C, together with R. japonica and R. longicrus, also formed a monophyletic group. Brown frogs related to Clades A and C were estimated to have dispersed from continental Asia to adjacent regions through multiple events. PMID- 17409720 TI - Ancient DNA analysis of the Japanese sea lion (Zalophus californianus japonicus Peters, 1866): preliminary results using mitochondrial control-region sequences. AB - In this study, we successfully extracted ancient DNA from skeletal remains of the Japanese sea lion-a species that is practically extinct-from archaeological sites and determined a partial sequence of its mitochondrial DNA control region. A molecular phylogenetic tree constructed by the neighbor-joining (NJ) method showed that the sequences from Japanese sea lions clustered together, with a high bootstrap value, and that this cluster was closest to the California sea lion cluster. The distinctly divergent cluster of Japanese sea lions reflected the morphological classification of these animals as a distinct species of the genus Zalophus; however, proximity to the California sea lion cluster simultaneously implied conformation with the traditional classification of these animals as a subspecies of Zalophus californianus. The average amount of nucleotide substitution between the Japanese and California sea lions was 7.02%. The Japanese and California sea lions were estimated to have diverged 2.2 million years ago, i.e., in the late Pliocene Epoch. This is the first report on a genetic analysis of the Japanese sea lion. PMID- 17409721 TI - A third photoreceptor-specific GRK found in the retina of Oryzias latipes (Japanese killifish). AB - We previously reported that the teleost fish medaka (Oryzias latipes, Japanese killifish), possesses two kinds of G protein-coupled receptor kinases (GRKs) in the retina with different localizations: GRK7 (OlGRK-C) in cones and GRK1 (OlGRK R1) in rods. To further clarify the diversity of teleost photoreceptor GRKs, we sought other medaka GRKs. We found an additional cDNA that encodes a second retina-specific GRK1 (OlGRK-R2). In situ hybridization experiments demonstrated that OlGRK-R2 mRNA is selectively expressed in rods. Sequence analysis of the Fugu rubripes genomic database unveiled a larger diversity of GRKs than previously expected. We also describe the light-dependent regulation of GRK1, a phenomenon that has not been found in other species. Immunocytochemical analysis indicated that OlGRK-R2 is localized in rod outer segments, independent of light condition. OlGRK-R1 is localized in the rod inner segments and synaptic termini of dark-adapted eyes, and moves to rod outer segments after light adaptation. Our studies suggest that the two medaka GRKs are not functionally redundant, and demonstrate a complicated light-dependent regulation of GRK1 in vivo. PMID- 17409722 TI - Expression of DMRT genes in the gonads of Rana rugosa during sex determination. AB - Sex is determined by various transcription factors. Dmrt1, a gene that contains the DM domain (Doublesex/Mab-3 DNA-binding motif), is known to be one of the critical factors required for testicular development in vertebrates. Recently, other DM domain-containing genes (DMRT genes) were isolated in mice and fish. In mice, three of the seven known DMRT genes are expressed in the embryonic gonad. In amphibians, however, only Dmrt1 and Dmrt4 have been cloned. To clarify the role(s) of DMRT genes in sex determination in vertebrates, the molecular cloning of amphibian DMRT genes would be very helpful. In this study, we isolated Dmrt2, 3, and -5 from the frog Rana rugosa and analyzed by RT-PCR their expression during sex determination. The analysis revealed that all three genes are expressed in the developing gonad/mesonephros complex during sex determination with no dimorphism. The results suggest that Dmrt2, -3, and -5 may be involved in sexual development in R. rugosa and that they may function in both testicular and ovarian differentiation. PMID- 17409723 TI - Resurrection of Staurois parvus from S. tuberilinguis from Borneo (Amphibia, Ranidae). AB - Two forms of Staurois that are differentiated by body size occur parapatrically in the Crocker Range, Sabah, Borneo. Analyses of a total of 1,499 bp of the mitochondrial cytochrome b, 12S rRNA, and 16S rRNA genes revealed that the two forms could be completely split genetically. The two forms could be also clearly differentiated morphologically, not only by snout-vent length but also by the relative sizes of snout, eye, and finger disk. Comparisons of the two forms with all known species of the genus revealed the large and small forms to be S. tuberilinguis and S. parvus, respectively. The latter species has long been synonymized with the former, but we here consider them to represent different species. PMID- 17409724 TI - A developmental basis for the Cambrian radiation. AB - As extant bilaterian phyla emerged during the Lower Cambrian, these clades acquired morphological features that separated them from their stem groups. At the same time, morphological variants on the body plan within a phylum emerged that we recognize as classes and subphyla. In many cases, the emergence of body plan variants within a phylum is associated with major changes in patterns of early regional specification. Subsequently these different patterns of regional specification were stabilized, probably because later developmental events depended on them. As a consequence, the frequency of new body plan variants involving early development declined in these lineages at later periods during their history. This hypothesis is explicated here by examining the process of early regional specification in the different subphyla of brachiopods and in pairs of species from the same subphylum belonging to different clades that originated during the Jurassic, Ordovician, and Lower Cambrian. PMID- 17409725 TI - Biogeography and karyotypes of freshwater planarians (Platyhelminthes, Tricladida, Paludicola) in southern Brazil. AB - In the Tricladida (Platyhelminthes), the incidence of different biotypes identified by several ploidy levels is very common. Planarians collected in the State of Rio Grande do Sul were identified using cytogenetics. Different species distributions were observed with respect to Rio Grande do Sul's geomorphology, which could have been caused by their different microhabitats. Girardia tigrina and G. anderlani consisted of diploid and triploid individuals, whereas G. schubarti showed diploids, triploids, and mixoploids; for all these species, individuals of different ploidies were sympatric. Only for diploid G. anderlani were B chromosomes observed. These B chromosomes seem to have an irregular segregational behavior during mitosis, and possibly also during meiosis. However the processes (e.g., selection, mutation) of maintaining 2n, 3n, and 2n/3n individuals within natural populations of G. schubarti remain to be clarified. PMID- 17409726 TI - Relationship between the lifestyle of a Siboglinid (Pogonophoran) polychaete, Oligobrachia mashikoi, and the total sulfide and nitrogen levels in its habitat. AB - A gutless polychaete of the family Siboglinidae, Oligobrachia mashikoi, known in the past as a beard worm of the group Pogonophora, inhabits Tsukumo Bay of the Noto Peninsula in the Sea of Japan. Photographs were taken of this polychaete projecting about one third of the length of its tentacles outside of its tube. The tube protruded several mm from the sea bottom. These are the first field photographs of beard worms. The trophosome of this beard worm harbors sulfur oxidizing bacteria. In fact, the muddy sediment where this worm inhabits smells slightly of hydrogen sulfide. Total sulfide levels, which can be an indicator of the generation of hydrogen sulfide gas, were measured at 10 locations in the bay. Furthermore, at the location which this species inhabits, the total sulfide levels in the vertical direction were determined. In addition, the total nitrogen levels, which can indicate the quantity of organic substances, were measured. The sediment inhabited by this worm was determined to have total sulfide levels of 0.24-0.39 mg/g dry mud, measured in the form of acid-volatile sulfide-sulfur. The total nitrogen levels were 1.0-1.5 microg/mg dry mud. These values suggest that the bottom of Tsukumo Bay has not been deteriorated by eutrophication. The levels were, however, highest in the surface layer of the sediment. These results suggest that hydrogen sulfide is generated in the surface of the sediment by sulfate-reducing bacteria, and that O. mashikoi appears to able to live in an environment that contains a slight amount of sulfide. PMID- 17409727 TI - Embryogenesis and expression profiles of charon and nodal-pathway genes in sinistral (Paralichthys olivaceus) and dextral (Verasper variegatus) flounders. AB - Although it is well known that flounder form external asymmetry by migration of one eye at metamorphosis, the control system that forms this asymmetry is unknown. To help elucidate this mechanism, we here describe the embryogenesis and expression profiles of the Nodal-pathway genes in the Japanese flounder, Paralichthys olivaceus. We also perform a comparative study of the laterality of the expression of these genes in sinistral (P. olivaceus) and dextral (Verasper variegatus) flounders. In P. olivaceus, Kupffer's vesicle forms at the 2-somite stage, after which left-sided expression of spaw starts at the 8-somite stage. Left-sided expression of pitx2 occurs in the gut field at the 15-somite to high pec stages, in the heart field at the 21-somite stage, and in the dorsal diencephalon at the 27-somite to high-pec stages. In response to left-sided pitx2 expression, the heart, gut, and diencephalon begin asymmetric organogenesis at the pharyngula (heart) and the long-pec (gut and diencephalon) stages, whereas the eyes do not show signs of asymmetry at these stages. In both sinistral and dextral flounders, the Nodal-pathway genes are expressed at the left side of the dorsal diencephalon and left lateral-plate mesoderm. Considering these data together with our previous finding that reversal of eye laterality occurs to some extent in the P. olivaceus mutant reversed, in which embryonic pitx2 expression is randomized, we propose that although the Nodal pathway seems to function to fix eye laterality, embryonic expression of these genes does not act as a direct positional cue for eye laterality. PMID- 17409728 TI - Cloning and expression of vacuolar proton-pumping ATPase subunits in the follicular epithelium of the bullfrog endolymphatic sac. AB - In an investigation aimed at clarifying the mechanism of crystal dissolution of the calcium carbonate lattice in otoconia (the mineral particles embedded in the otolithic membrane) of the endolymphatic sac (ELS) of the bullfrog, cDNAs encoding the A- and E-subunits of bullfrog vacuolar proton-pumping ATPase (V ATPase) were cloned and sequenced. The cDNA of the A-subunit consisted of an 11 bp 5'-untranslated region (UTR), a 1,854-bp open reading frame (ORF) encoding a protein comprising 617 amino acids with a calculated molecular mass of 68,168 Da, and a 248-bp 3'-UTR followed by a poly(A) tail. The cDNA of the E-subunit consisted of a 72-bp 5'-UTR, a 681-bp ORF encoding a protein of 226 amino acids with a calculated molecular mass of 26,020 Da, and a 799-bp 3'-UTR followed by a poly(A) tail. Western blot and immunofluorescence analyses using specific anti peptide antisera against the V-ATPase A- and E-subunits revealed that these subunits were present in the ELS, urinary bladder, skin, testes, and kidneys. In the ELS, positive cells were scattered in the follicular epithelium which, as revealed by electron microscopy, corresponds to the location of mitochondria-rich cells. These findings suggest that V-ATPase, including the A- and E-subunits, exists in mitochondria-rich cells of the ELS, which might be involved in dissolution of the calcium carbonate crystals in the lumen of the ELS. PMID- 17409729 TI - Inbred strains of brine shrimp derived from Artemia franciscana: lineage, RAPD analysis, life span, reproductive traits and mode, adaptation, and tolerance to salinity changes. AB - Inbred strains of the brine shrimp were developed from dry dormant cysts of wild type Artemia franciscana produced in the Great Salt Lake, U.S.A. The established strains were named GSL2, 4, and 7. They were raised in 2% natural sea salt solution at 28 degrees C under a long-day condition, and fed on food sold for Artemia. Ovoviviparous offspring (free-swimming nauplii) in each brood derived from full sib (sister x brother) matings were used for succeeding generations. The ordinal number of the filial generation increased at a rate of ten generations per year. The number was over 60, and the lineage was recorded. Random amplified polymorphic DNA (RAPD) analyses of the inbred strains revealed the uniqueness, homogeneity, and genetic similarity among them. Their life span, the time required to become sexually mature, brood size, mode of reproduction, and adaptation and tolerance to salinity changes were investigated. The inbred strains usually released free-swimming nauplii rather than spawning encysted gastrulae (dormant cysts). On the other hand, the opposite results were obtained from wild-type Artemia under the same conditions. Both adults and nauplii of the inbred strains appeared to be less adaptive and less tolerant to salinity changes compared to those of the wild type. The established inbred strains should provide a wider and deeper scope for Artemia biology in particular, and the life sciences in general. PMID- 17409730 TI - Phylogenetic relationships within the genus Jesogammarus (Crustacea, Amphipoda, Anisogammaridae) deduced from mitochondrial COI and 12S sequences. AB - The genus Jesogammarus contains 16 species in two subgenera, Jesogammarus and Annanogammarus. To examine relationships among species in the genus, a molecular phylogenetic study including eight species of the former subgenus and four of the latter was conducted using partial DNA sequences of the mitochondrial COI and 12S rRNA genes. MP, NJ, and ML trees based on the combined COI and 12S data indicated monophyly of the subgenus Annanogammarus, though the monophyly of Jesogammarus was left unresolved. Consistent with few morphological differences, Jesogammarus (A.) naritai and J. (A.) suwaensis showed low genetic differentiation and did not show reciprocal monophyly, which suggests a close affinity of these taxa. PMID- 17409731 TI - Ecology and reproductive patterns of the agamid lizard Japalura swinhonis on an east Asian island, with comments on the small clutch sizes of island lizards. AB - I describe the habitat use, diet, and the male and female reproductive cycles of Japalura swinhonis, an oviparous agamid lizard inhabiting Orchid Island, a tropical island off the southeastern coast of Taiwan. Ninety percent of lizards (n=126) were observed on tree trunks or at the forest edge. The diet of J. swinhonis on Orchid Island consisted mostly of hymenopterans (53.33%) and orthopterans (16.67%). The mean snout-vent length (SVL) of adult males was 74.58 (n=89) and that of females was 69.31 (n=37) mm. Females exhibited a long vitellogenic period from November to February, with parturition occurring from March to October. The onset of vitellogenesis did not correlate with the mass of the female fat bodies. Females produced two to five eggs per clutch, and clutch size was not correlated with SVL. Two clutches were recorded during a single year in some individuals. Clutch size in J. swinhonis was compared with that in other Japalura species. Clutch sizes of Japalura species are larger in mainland China than on insular Taiwan. Clutch size is also mainly affected by environmental constraints, and smaller clutch sizes are probably affected by predators on Orchid Island. PMID- 17409732 TI - Allozyme variation in the camaenid tree snails Amphidromus atricallosus (Gould, 1843) and A. inversus (Muller, 1774). AB - We examined allozyme variation in two camaenid tree snails, Amphidromus atricallosus and A. inversus, across two principal regions of Thailand and from Singapore, plus for A. inversus, one site in peninsular Malaysia. Using horizontal starch gel electrophoresis, 13 allozyme loci (11 polymorphic) were screened for A. atricallosus and 18 (5 polymorphic) for A. inversus. Heterozygosity was higher in A. atricallosus (Hexp=0.018-0.201, mean=0.085) than in A. inversus (Hexp=0-0.023, mean= 0.002). Genetic heterogeneity among samples was higher in A. inversus (Fst=0.965) than in A. atricallosus (Fst=0.781). Within A. atricallosus, populations were more differentiated in southern Thailand (Fst=0.551) than in eastern Thailand (Fst=0.144). The high Fst and low Hexp in populations of A. inversus suggest that this species is likely to have experienced a series of strong bottlenecks, perhaps occurring chiefly on offshore continental-shelf islands. The low Fst values of A. atricallosus in eastern Thailand suggest frequent gene flows among populations in this region. The southern and eastern samples of A. atricallosus exhibited fixed allele differences at four loci and great genetic distance (Nei's D=0.485-0.946), suggesting that these two samples may actually represent, or else be evolving into, separate species. PMID- 17409733 TI - Unmasking Rana okinavana Boettger, 1895 from the Ryukyus, Japan (Amphibia: Anura: Ranidae). AB - Examination of the lectotype and a paralectotype of Rana okinavana Boettger, 1895 revealed that the species is not a brown frog of the subgenus Rana, occurring in the middle group of the Ryukyu Archipelago, but is identical with a frog of the subgenus Nidirana from the southern group of the Archipelago and Taiwan, now called R. psaltes Kuramoto, 1985. The type locality of R. okinavana given in the original description, Okinawa of the middle Ryukyus, is highly doubtful and should be somewhere in the Yaeyama Islands of the southern Ryukyus. The name R. psaltes is relegated to a subjective junior synonym of R. okinavana Boettger, 1895, while the brown frog of the subgenus Rana from the northern Ryukyus requires a replacement name. PMID- 17409734 TI - A model for the involvement of neural cell adhesion molecules in stress-related mood disorders. AB - Critical interactions between genetic and environmental factors -- among which stress is one of the most potent non-genomic factors -- are involved in the development of mood disorders. Intensive work during the past decade has led to the proposal of the network hypothesis of depression [Castren E: Nat Rev Neurosci 2005;6:241-246]. In contrast to the earlier chemical hypothesis of depression that emphasized neurochemical imbalance as the cause of depression, the network hypothesis proposes that problems in information processing within relevant neural networks might underlie mood disorders. Clinical and preclinical evidence supporting this hypothesis are mainly based on observations from depressed patients and animal stress models indicating atrophy (with basic research pointing at structural remodeling and decreased neurogenesis as underlying mechanisms) and malfunctioning of the hippocampus and prefrontal cortex, as well as the ability of antidepressant treatments to have the opposite effects. A great research effort is devoted to identify the molecular mechanisms that are responsible for the network effects of depression and antidepressant actions, with a great deal of evidence pointing at a key role of neurotrophins (notably the brain-derived neurotrophic factor) and other growth factors. In this review, we present evidence that implicates alterations in the levels of the neural cell adhesion molecules of the immunoglobulin superfamily, NCAM and L1, among the mechanisms contributing to stress-related mood disorders and, potentially, in antidepressant action. PMID- 17409735 TI - The limbic system in Mammalian brain evolution. AB - Previous accounts of mammalian brain allometry have relied largely on data from primates, insectivores and bats. Here we examine scaling of brain structures in carnivores, ungulates, xenarthrans and sirenians, taxa chosen to maximize potential olfactory and limbic system variability. The data were compared to known scaling of the same structures in bats, insectivores and primates. Fundamental patterns in brain scaling were similar across all taxa. Marine mammals with reduced olfactory bulbs also had reduced limbic systems overall, particularly in those structures receiving direct olfactory input. In all species, a limbic factor with olfactory and non-olfactory components was observed. Primates, insectivores, ungulate and marine mammals collectively demonstrate an inverse relationship between isocortex and limbic volumes, but terrestrial carnivores have high relative volumes of both, and bats low relative volumes of both. We discuss developmental processes that may provide the mechanistic bases for understanding these findings. PMID- 17409736 TI - Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cell markers. AB - We report the isolation of a population of immature dental pulp stem cells (IDPSC), which express embryonic stem cell markers Oct-4, Nanog, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81 as well as several other mesenchymal stem cell markers during at least 25 passages while maintaining the normal karyotype and the rate of expansion characteristic of stem cells. The expression of these markers was maintained in subclones obtained from these cells. Moreover, in vitrothese cells can be induced to undergo uniform differentiation into smooth and skeletal muscles, neurons, cartilage, and bone under chemically defined culture conditions. After in vivo transplantation of these cells into immunocompromised mice, they showed dense engraftment in various tissues. The relative ease of recovery and the expression profiles of various markers justify further exploration of IDPSC for clinical therapy. PMID- 17409737 TI - Applicability of the dyes CFSE, CM-DiI and PKH26 for tracking of human preadipocytes to evaluate adipose tissue engineering. AB - Adipose tissue engineering with preadipocyte-loaded scaffolds requires adequate tracking of preadipocytes to allow evaluation and quantification of cell proliferation, expansion and differentiation in three-dimensional systems. To differentiate between graft and host cells, labeling of preadipocytes before implantation and tracking of these cells until harvest would be useful. Immunohistochemistry enables the differentiation between cells of different species but is time-consuming, expensive, elaborate, and not applicable for autologous transplantation. So far, there is no published method to use externally applied dyes for tracking of human preadipocytes in adipose tissue engineering. We tested the cell dyes PKH26, CM-DiI, and CFSE to analyze their applicability for labeling human preadipocytes. CM-DiI had toxicity levels of 45 70%, while 3-4% proliferating cells were stained on day 35. CFSE revealed clear cytoplasmic coloring in proliferating cells with 5-6% stained cells after 35 days and toxicity ranging from 55 to 90% dead cells. PKH26 demonstrates lowest levels of toxicity and best labeling results after 4 weeks in proliferating preadipocytes in monolayer. Although none of the dyes showed long-lasting labeling during proliferation, all three dyes demonstrated permanent staining in differentiated cells. The results reveal the problems of preadipocyte tracking with fluorescent dyes but justify the dye application for limited time periods. PMID- 17409738 TI - Histomorphometric analysis of tissue responses to bioactive glass implants in critical defects in rat calvaria. AB - The aim of this study was to evaluate the osteogenic behavior of two chemically similar bioactive glass products (Biogranand Perioglas) implanted in critical bone defects in rat calvaria. Thirty-six transfixed bone defects of 8 mm diameter were made surgically in adult male Wistar rats. The animals were distributed equally into three groups: Biogran (GI), Perioglas (GII) and without implant material (control; GIII). The morphology and composition of both bioactive glasses were analyzed by scanning electron microscopy and energy-dispersive spectrometry. Tissue specimens were analyzed at the biological time points of 15, 30 and 60 days by optical microscopy and morphometry, demonstrating biocompatibility for the tested materials with moderate chronic inflammation involving their particles. Bone neoformation resulted only as a reparative reaction to an intentionally produced defect and was limited to the defect's edges. No statistically significant differences among the groups were observed. At the scar interstice, abundant deposits of collagenous fibers enveloping the particles were noted. The present results indicated that the bioactive glasses, under the experimental conditions analyzed, did not show osteogenic behavior. PMID- 17409739 TI - Changes in bone microarchitecture and bone mineral density following experimental osteonecrosis of the hip in rabbits. AB - BACKGROUND: Osteonecrosis of the femoral head is a common disorder which can lead to hip joint destruction usually necessitating total hip replacement. METHODS: Quantitative micro-computed tomography, digital radiography and histology were used to characterize changes in bone microarchitecture and bone mineral density during the repair of the osteonecrotic femoral head as well as during the development of secondary osteoarthritis in the ipsilateral acetabulum. Osteonecrosis was induced surgically in 17 adult, male rabbits and the contralateral side was used as control. RESULTS: At 4 weeks no changes in microarchitecture in the femoral head nor in the acetabulum were found. At 6 months the repair process led to an increase in bone mass in the trabecular region of the femoral head. However, a decrease in volumetric bone mineral density and an increase in apparent porosity were seen in the compact subchondral and cortical region of the osteonecrotic femoral head. At 6 months the subchondral bone of the osteoarthritic ipsilateral acetabulum was thicker, but had a lower volumetric bone mineral density and a higher apparent porosity. CONCLUSION: Resorption of necrotic compact bone may weaken the structural properties of the femoral head. Moreover, remodeling and resorption of subchondral bone may play a critical role in the disease process of osteoarthritis. PMID- 17409740 TI - Digital measurements: a different approach to evaluate bone formation. A technical report. AB - In this study a digital measurement technique has been proposed to quantify bone formation on histological images. Two standard parietal defects were created in 30 adult rabbits. The animals were divided into six groups. Four animals of each group were randomly chosen as experimental group in which osteopontin-coated hydroxyapatite (OPN-HA) and hydroxyapatite (HA) were inserted alternatively in created defects. To observe the spontaneous healing process of defects, one animal of each group was used as control animal and these created defects did not receive any implants. The animals were sacrificed after 1, 2, 6, 12, 18 and 30 weeks. The histological sections were magnified (x100) and scanned digitally. The newly formed bone surfaces within the healing area were indicated and quantified by means of Adobe Photoshop 7 software. This measuring technique was found to be reliable and reproducible. The results of this study show no significant differences in bone formation between the OPN-HA and non-coated HA defects, although a significant difference in bone formation was measured at the margins of the defects treated with OPN-HA. PMID- 17409741 TI - Comparison of polyester scaffolds for bioengineered intestinal mucosa. AB - INTRODUCTION: Biodegradable polyester scaffolds have proven useful for growing neointestinal tissue equivalents both in vitro and in vivo. These scaffolds allow cells to attach and grow in a 3-dimensional space while nutrient flow is maintained throughout the matrix. The purpose of this study was to evaluate different biopolymer constructs and to determine mucosal engraftment rates and mucosal morphology. HYPOTHESIS: We hypothesized that different biopolymer constructs may vary in their ability to provide a good scaffolding onto which intestinal stem cell organoids may be engrafted. STUDY DESIGN: Eight different microporous biodegradable polymer tubes composed of polyglycolic acid (PGA), polylactic acid, or a combination of both, using different fabrication techniques were seeded with intestinal stem cell clusters obtained from neonatal rats. Three different seeded polymer constructs were subsequently placed into the omentum of syngeneic adult recipient rats (n = 8). Neointestinal grafts were harvested 4 weeks after implantation. Polymers were microscopically evaluated for the presence of mucosal growth, morphology, scar formation and residual polymer. RESULTS: Mucosal engraftment was observed in 7 out of 8 of the polymer constructs. A maximal surface area engraftment of 36% (range 5-36%) was seen on nonwoven, randomly entangled, small fiber PGA mesh coated with aerosolized 5% poly-L-lactic acid. Villous and crypt development, morphology and created surface area were best on PGA nonwoven mesh constructs treated with poly-L-lactic acid. Electrospun microfiber PGA had poor overall engraftment with little or no crypt or villous formation. CONCLUSION: Intestinal organoids can be engrafted onto biodegradable polyester scaffoldings with restitution of an intestinal mucosal layer. Variability in polymer composition, processing techniques and material properties (fiber size, luminal dimensions and pore size) affect engraftment success. Future material refinements should lead to improvements in the development of a tissue-engineered intestine. PMID- 17409742 TI - Three-dimensional surface structure of macrophages in fetal and adult mouse liver: an immunohistochemical light microscopic study. AB - Using 100-microm-thick paraffin sections stained by F4/80 antibody, the three dimensional surface morphology of macrophages in fetal and adult livers was examined by conventional light microscope equipped with a computer-controlled z axis stepping motor. Hematopoietic macrophages in fetal livers were located in the center of erythroid cell clusters, forming cell sockets which consisted of two different kinds of projections. The primary cytoplasmic processes were membranous projections and the secondary processes were finger-like projections extending from the primary processes. Erythroids in the cell sockets were linearly arranged on the macrophage surface. Adult sinusoidal macrophages possessed a few pseudopod-like processes, ridge-like profiles and numerous ruffle or spike-like processes, and cell contact with neighboring macrophages could be recognized. Compared to confocal laser scanning microscopy and scanning electron microscopy, this study provided information about cell surface structures at reasonably low cost, although the resolution was limited in z axis due to the stepping interval. PMID- 17409743 TI - ALCAM (CD166) is a surface marker for early murine cardiomyocytes. AB - ALCAM (activated leukocyte cell adhesion molecule, CD166) belongs to the immunoglobulin superfamily and is involved in axon guidance, hematopoiesis, immune response and tumor metastasis. During embryogenesis, mRNA encoding ALCAM was expressed in the cardiac crescent and the neural groove at embryonic day (E) 7.75 and predominately in the tubular heart at E8.5. A newly generated monoclonal antibody against the ALCAM molecule (ALC-48) exclusively stained cardiomyocytes at E8.25-10.5. However, ALCAM expression was lost by cardiomyocytes by E12.5 and its expression shifts to a variety of organs during later stages. ALCAM was found to be a prominent surface marker for cardiomyocytes in early embryonic hearts. The transient expression of ALCAM during early developmental stages marks specific developmental stages in cardiomyocyte differentiation. PMID- 17409744 TI - Aquaporin 1: examination of its expression and localization in normal human skeletal muscle tissue. AB - To examine aquaporin 1 (AQP1) expression in skeletal muscle tissue precisely, we performed reverse transcription-polymerase chain reaction (RT-PCR) at RNA level and immunoblot analysis, immunohistochemistry and immunoelectron microscopy at protein level. The RT-PCR study of total RNA from normal human skeletal muscle showed a strong single band of AQP1. At the protein level we used two commercially available antibodies, both of which recognize the cytoplasmic domain of the AQP1 molecule. One antibody gave positive results. Immunoblot of muscle extract showed a 30-kDa band protein, the molecular weight of which corresponded to that of AQP1. Immunohistochemically, AQP1 was immunostained at the myofiber surface both in type 1 and type 2 myofibers with almost the same intensity, and its staining pattern was rather diffuse and irregular compared with that of the anti-dystrophin antibody. The endomysial endothelial cells were also immunolabeled. Immunoelectron microscopy revealed that the immunogold particles indicating the presence of the AQP1 molecule were present along the inside surface of the muscle plasma membrane. However, another antibody showed negative results except for the endomysial endothelial cells which were positively stained. We drew the conclusion that AQP1 is expressed at the endomysial capillary endothelial cell and further AQP1 may be expressed at the human skeletal myofiber plasma membrane. PMID- 17409745 TI - Immunohistochemical study of somatotrophs in pituitary pars distalis of male viscacha (Lagostomus maximus maximus) in relation to the gonadal activity. AB - Somatotrophs were identified and quantified in pituitary pars distalis of male viscachas (Lagostomus maximus maximus) during the annual reproductive cycle, after the administration of melatonin, after castration and in different growth stages by immunohistochemistry and morphometric analysis. In adult male viscachas, the somatotrophs were distributed throughout the pars distalis during the reproductive cycle. They were oval, pyramidal or round in shape with a large round nucleus. The percentage immunopositive area, the major cellular diameter and the number of cells decreased during the gonadal regression period in relation to the values found in the reproductive period. The administration of melatonin did not provoke any variations of the morphometric parameters studied. On the contrary, a significant decrease in the percentage immunopositive area, in the major cellular diameter and in the number of somatotrophs in castrated viscachas was observed. The study of different growth stages showed that these morphometric parameters increased from immature to adult animals in the reproductive period. The results obtained suggested that the variations of the morphometric parameters of somatotrophs are more related to the gonadal development and activity than to a direct effect of melatonin. PMID- 17409746 TI - The prevention of vocal fold scarring using autologous adipose tissue-derived stromal cells. AB - Prevention and treatment of vocal fold scarring and atrophy remain challenging. The aim of this study was to treat injured vocal folds using autologous adipose tissue-derived stromal cells (ADSCs) and evaluate the ability to prevent vocal fold scarring and atrophy by ADSCs in a canine animal model. Ten adult dogs were used for this experiment. ADSCs from the adipose tissue from the inguinal area were isolated and cultured in all dogs. Immediately after being mixed with atelocollagen, the ADSCs (1-3 x 10(6)) were injected into the right vocal fold of each animal, using a syringe with a 23-gauge needle. As a control, atelocollagen was injected into the left vocal fold of the same dog. The effects of the prevention of vocal fold scarring and atrophy were measured by morphological and histological assessment. At 8 weeks, there was a difference in granuloma and atrophic changes between the ADSC-injected and control sides in the majority of the dogs. This difference continued to be present at the 24 weeks' follow-up. On histopathologic examination, a large number of cells labeled with a fluorochrome were observed in ADSC-injected vocal folds 8 weeks after the initial treatment. This study demonstrates the multipotential ability of ADSCs in the regeneration of injured vocal folds. Injecting ADSCs into a damaged vocal fold appears to be useful in preventing vocal fold scarring and atrophy 24 weeks after initial damage. PMID- 17409747 TI - Anatomical definition of the subglottic region. AB - BACKGROUND: Due to laryngeal neoplasia, as well as infectious and autoimmune diseases, the subglottic region is of great clinical relevance. However, descriptions of the subglottic structures are inconsistent. The aim of our study was to present a precise analysis of the subglottic region and derive functional and clinical conclusions. METHODS: Histological, histochemical and immunohistochemical investigations as well as scanning electron microscopy were performed and combined with injection techniques applied to the subglottic region of the larynges of 33 body donors. RESULTS: The three-dimensional extensions of the subglottic region were newly defined: the inferior arcuate line of the vocal cord was defined as the cranial border. The lower margin of the cricoid is the caudal border. Craniolaterally, the elastic cone and, further caudally, the cricoid form the border. Therefore, the definition presented comprises heretofore unnamed ventral and dorsal parts of the lower larynx. The subglottic region can be described as cylindrical, becoming smaller in the cranial direction, following the elastic cone. The ventral boundary is formed by the median part of the cricothyroid ligament (ligamentum conicum), the dorsal part by the cricoid cartilage. The walls of the subglottis are divided into three or four layers composed of collagenous and elastic fibres in which seromucous glands are embedded. Subglottic blood vessels including a tight subepithelial capillary plexus were delineated. CONCLUSION: The new definition of the subglottic extensions presented is helpful and essential for precise laryngeal tumour classification. The results indicate that the boundaries are unlikely to counteract tumour progression in the subglottis. Furthermore, the findings suggest that these structures contribute to temperature regulation of breath, protection against inflammation as well as breath frequency and depth-dependent mucus secretion. PMID- 17409748 TI - Efficacy and safety of galantamine in patients with dementia with Lewy bodies: a 24-week open-label study. AB - BACKGROUND: Dementia with Lewy bodies (DLB) is a common dementia of the elderly. A significant cholinergic deficit has been demonstrated that may be responsive to treatment by cholinesterase inhibitors (ChEIs). METHODS: A 24-week, open-label study was designed to assess the efficacy and safety of a ChEI, galantamine, in 50 patients with DLB. RESULTS: This study showed beneficial effects with galantamine in 2 of the 3 primary efficacy parameters. The scores on the Neuropsychiatric Inventory (NPI-12) improved by 8.24 points from baseline (p = 0.01) especially in visual hallucinations and nighttime behaviors (p = 0.004). The scores on the Clinician's Global Impression of Change improved by 0.5 points from baseline (p = 0.01). The third primary efficacy parameter, the Cognitive Drug Research Computerized Cognitive Assessment System, was unchanged from baseline. Adverse events were generally mild and transient. CONCLUSION: Galantamine appears to be an effective and safe therapy for patients with DLB. PMID- 17409749 TI - Can Klinefelter patients be female? PMID- 17409750 TI - The application of an enamel matrix protein derivative (Emdogain) in regenerative periodontal therapy: a review. AB - Regenerative periodontal therapy aims at reconstitution of the lost periodontal structures such as new formation of root cementum, periodontal ligament and alveolar bone. Findings from basic research indicate that enamel matrix protein derivative (EMD) has a key role in periodontal wound healing. Histological results from animal and human studies have shown that treatment with EMD promotes periodontal regeneration. Moreover, clinical studies have indicated that treatment with EMD positively influences periodontal wound healing in humans. This review aims to present an overview of evidence-based clinical indications for regenerative therapy with EMD. PMID- 17409751 TI - Low back ache treatment with botulinum neurotoxin type A. Local experience in Kuwait. AB - OBJECTIVE: To investigate the efficacy, safety and tolerability of paraspinal administration of botulinum neurotoxin type A (BoNT-A) in patients with chronic low back ache (LBA). SUBJECTS AND METHODS: Eight patients with chronic LBA were injected with BoNT-A at three sites on either side of lumbar paraspinal muscles. The patients rated their pain intensity using a visual analogue scale (VAS) from 0 to 10, and our physiotherapist assessed the paraspinal muscle spasm using a functional scale (FS) from 0 to 5. The improvement in both VAS and FS 30, 60 and 90 days after BoNT-A from baseline was analyzed separately using paired t test. The correlation between the muscle spasm (FS) and pain relief (VAS) was analyzed using the Spearman's rank correlation coefficient test. The level of statistical significance was p < 0.05. RESULTS: Five (63%) patients showed a remarkable recovery in VAS and FS, whereas 2 (25%) patients showed improvement only in FS. Statistically significant improvement was achieved in VAS and FS independently 30 days (p < 0.02 and p < 0.02, respectively), 60 days (p < 0.01 and p < 0.001, respectively) and 90 days (p < 0.02 and p < 0.001, respectively) after treatment. Pain relief started early and it was independent of relief of muscle spasm 30 days after treatment (r = 0.685; p > 0.05). CONCLUSION: With this limited study, we have demonstrated that the paraspinal injection of BoNT-A is effective in relieving chronic LBA without producing side effects. The injection is an easy procedure, well tolerated and did not require anesthesia or imaging technique. PMID- 17409752 TI - Risk factors for mortality of nosocomial bacteraemia in intensive care units. AB - OBJECTIVE: The aim of this study was to follow critically ill patients prospectively in intensive care units (ICUs) to determine risk factors for mortality and outcome associated with nosocomial bacteraemia (NB). SUBJECTS AND METHODS: A case-control study of 176 patients was conducted to identify the risk factors for mortality of NB in ICU patients. The study was performed in emergency, surgical and general surgical ICUs with 23 beds during a 15-month period. A total of 1,450 patients were admitted to the ICUs during the study period. The USA Center for Disease Control and Prevention definitions were used to diagnose nosocomial infections. Nosocomial bacteraemia was defined as the isolation of one or more organisms from blood cultures taken at least 48 h after admission, which were not related to a problem present on admission. An assessment of whether the isolated organisms represented true bacteraemia rather than contamination was made by clinical or laboratory evidence of infection. RESULTS: A total of 214 bacteraemia episodes were found in the 176 patients (64 female, 112 male; 51.3 +/- 21.3 years old), 90 of whom died and 86 survived. The bacteraemia rate was 12.1%. The most common etiological agents of bacteraemia were Klebsiella pneumoniae: 46 (21.5%), methicillin-resistant Staphylococcus aureus: 46 (21.5%), Pseudomonas aeruginosa: 32 (14.9%), and Escherichia coli: 20 (9.3%). Multivariate analysis showed that the requirement of mechanical ventilation for more than 7 days (p < 0.001), total parenteral nutrition (p = 0.034), inotropic drug (p < 0.001), and increased creatinine level (p = 0.034) were independent risk factors for mortality of NB in ICUs. CONCLUSIONS: Nosocomial infections caused by Gram-negative bacteria continue to be one of the major sources of morbidity and mortality. PMID- 17409753 TI - Cardiac memory versus likelihood of ischemic heart disease in hypertensive patients with ventricular repolarization abnormalities after repetitive uniform ventricular extrasystoles. AB - OBJECTIVES: To investigate the value of T-wave inversion after premature ventricular complexes (PVCs) for the prediction of hypertensive patients with a likelihood of ischemic heart disease (IHD). PATIENTS AND METHODS: Ambulatory 24 hour ECG, ambulatory 24-hour blood pressure (BP) recording, echocardiography, treadmill exercise ECG test and thallium-201 scintigraphy were performed for 210 hypertensive patients. RESULTS: Predictive indices showed that persistent symmetrical T-wave inversion after repetitive uniform PVCs equaling 30 min can be considered an indicator for the prediction of IHD in hypertensive patients as sensitivity was 80%, specificity: 73%, accuracy: 78%, positive predictive value: 83%, and negative predictive value: 69%. kappa coefficient value (kappa) indicated that there was good agreement between changes in ventricular repolarization after PVCs and stress thallium scintigraphy (kappa = 0.897). Receiver-operating characteristic curve data showed that the ideal cutoff value of T-wave inversion voltage to predict IHD in hypertensive patients was 2.2 mV (sensitivity: 66% and false positive: 35%, and area under curve: 0.716), but the ideal cutoff value of time for resolution of T-wave inversion was 34 min (sensitivity: 83% and false positive: 18%, and area under curve: 0.873). Multivariate analysis revealed that T-wave inversion after repetitive uniform PVCs was significantly associated with non-sustained ventricular tachycardia, decreased parasympathetic activity, increased sympathetic activity, morning BP surge, systolic BP load >50% and left ventricular diastolic dysfunction status. CONCLUSION: The data show that persistent symmetrical T-wave changes following resumption of sinus rhythm after repetitive uniform PVCs are an independent variable for the prediction of IHD in hypertensive patients. PMID- 17409754 TI - Classifying intertrochanteric fractures of the proximal femur: does experience matter? AB - OBJECTIVE: This prospective study aimed to evaluate inter-observer agreement of the AO/OTA and Evans/Jensen classification systems with an emphasis on the effect of reviewer experience. In addition, the level of agreement in the determination of fracture stability across varying levels of experience was evaluated. SUBJECT AND METHODS: A group of 12 reviewers (6 surgeons, 3 senior residents, 3 junior residents) independently assessed 56 radiographs of intertrochanteric hip fractures and classified them using the AO/OTA and Evans/Jensen systems. Reviewers further assessed the stability of the fracture patterns. RESULTS: Overall, higher agreement was obtained for the AO/OTA classification than with the Evans/Jensen classification. However, neither classification system met acceptable thresholds for reliability. Surgeons were unable to reliability determine if fractures were stable or unstable. Increased experience improved AO/OTA classification reliability and assessment of stability but decreased agreement in the Evans/Jensen classification. CONCLUSION: The AO/OTA classification should be used in favor of the Evans/Jensen classification whenever possible. Our findings suggest that surgeons' perceptions about stability vary to a significant extent thereby necessitating clear definitions of stability. PMID- 17409755 TI - Inhibition of angiotensin-converting enzyme activity by a partially purified fraction of Gynura procumbens in spontaneously hypertensive rats. AB - OBJECTIVES: To investigate the hypotensive and angiotensin-converting enzyme (ACE) inhibitory activities of a partially purified fraction (FA-I) of the leaves of Gynura procumbens and to qualitatively analyse the putative compounds present in the fraction. MATERIALS AND METHODS: The hypotensive effect of FA-I was tested in both spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto rats (WKY) by an intravenous administration of 0-10 mg/kg of the FA-I. Administration of captopril (20 microg/kg) served as the control. In vitro 0.0-2.0 mg/ml FA-I was added to a mixture of ACE and hippuryl-L-histidyl-L-leucine and assayed by a modification of the colourimetric method of Hurst and Lovell-Smith. All blood pressure measurements were monitored by the Macintosh MacLab set-up. ACE activity was measured by an in vitro assay in which the enzymatic cleavage of hippuryl-L histidyl-L-leucine to form histidyl-leucine and hippurate was determined colourimetrically by a cyanuric chloride/dioxane reagent. RESULTS: The FA-I produced a marked dose-dependent reduction in mean arterial pressure (MAP) in SHR and WKY rats, with an ED(50) of 1.09 and 1.05 mg/kg, respectively (p < 0.01). Furthermore, FA-I at 10 mg/kg strongly inhibited the angiotensin I-induced rise in MAP (p < 0.01). This response was comparable to that of captopril at 20 microg/kg. In the in vitro assay, ACE activity was inhibited with an IC(50) of 0.8 mg/ml. The qualitative phytochemical analysis of FA-I indicated the presence of glycoconjugates and peptides. CONCLUSION: These results suggest that the hypotensive effect of G. procumbens may be due, in part, to the glycoconjugated or peptidal substances found in FA-I that exhibit an inhibitory effect on ACE. PMID- 17409756 TI - Paragangliomas of the head and neck. AB - OBJECTIVES: The purpose of this study was to determine the clinical characteristics of carotid body tumors and to analyze the benefits of different treatment modalities. SUBJECTS AND METHODS: The clinical records of 13 patients with paragangliomas treated during the period of 1995-2005 were reviewed for age at diagnosis, sex, initial symptoms, duration of symptoms, evidence of secretory function, family history, radiographic studies, pre- and posttreatment cranial nerve deficits, multicentric tumors, angiography, operative procedures and findings, and complications. The diagnoses were established by computerized tomography and magnetic resonance imaging scanning. Angiography was performed on 3 patients with carotid body tumor. RESULTS: All of the patients were followed up for a duration ranging from 16 months to 9 years. The median follow-up was 5 years. Of all 18 head and neck tumors confirmed in 13 patients, 16 were in the carotid body, 1 in the vagus nerve and 1 in the middle ear cleft. Four patients had multiple paragangliomas, 2 bilateral synchronous and 1 bilateral asynchronous paragangliomas. The remaining patient had 3 asynchronous paragangliomas, i.e. left jugulotympanic, right carotid body and left vagal paraganglioma. All patients underwent successful surgical resection of the tumor after appropriate preoperative investigation. CONCLUSION: This study shows that surgical treatment is acceptably safe and effective in treating these neoplasms. PMID- 17409757 TI - Development, validation and application of tandem mass spectrometry for screening of inborn metabolic disorders in Kuwaiti infants. AB - OBJECTIVE: The aim of this work was to develop a specific and validated tandem mass spectrometric (MS/MS) method for screening of amino acidopathies, organic acidurias, urea cycle disorders and fatty acid oxidation defects in Kuwaiti newborns and sick infants. MATERIALS AND METHODS: A total of 1,520 blood samples were tested for inborn metabolic disorders in Kuwaiti newborns and sick infants. Positive electrospray MS/MS was used to measure diagnostic acylcarnitines and amino acids in blood spots after simple extraction and derivatization procedures. Validation and stability studies were conducted using control blood samples supplemented with known concentrations of the diagnostic amino acids or acylcarnitines. Reference and cutoff levels of the diagnostic metabolites were determined in a group of 500 normal Kuwaiti babies for quantitative evaluation. RESULTS: Of the 1,520 samples, 32 were positive newborn cases and 27 positive symptomatic infants. For the validation studies, the range of relative standard deviation was 2.6-14.7%, whereas the range of the percent deviation from nominal concentrations was -23.0 to +25.0 of the diagnostic metabolites. Stability studies indicated appropriate stability of the diagnostic amino acids and acylcarnitines in dried blood spots stored at 22 +/- 1 degrees C and relative humidity of 50-60%. CONCLUSIONS: Tandem mass spectrometry can significantly contribute to a newborn screening program as a fast and highly specific diagnostic technique for screening of a broad range of inborn metabolic disorders. PMID- 17409758 TI - Correlation between increased serum sFas levels and microalbuminuria in type 1 diabetic patients. AB - OBJECTIVE: The aim of this study was to elucidate if apoptosis dysregulation is present in type 1 diabetic patients with microalbuminuria. SUBJECTS AND METHODS: The following variables were determined in 29 type 1 diabetic patients: the duration of diabetes, soluble Fas (sFas), Bcl-2, hemoglobin A(1c) levels, glomerular filtration rate (GFR) and microalbuminuria, using the urine albumin to urine creatinine ratio (ACR). Age and gender were assessed and patients were categorized into two groups, according to their ACR: the microalbuminuric (MA) group with an ACR > or =30 mg/g, and the normoalbuminuric (NA) group with an ACR <30 mg/g. RESULTS: The differences between the two groups regarding sFas, Bcl-2 and GFR were not statistically significant. However, in the MA group, a significant positive relationship between sFas and ACR was observed (r = 0.736, p = 0.015). Dividing patients into two subgroups--mild versus severe (ACR > or =150 mg/g) microalbuminuric patients--significant differences in sFas (60.4 vs. 87.2 pg/ml; p = 0.047) and GFR (113 vs. 69.5 ml min(-1) 1.73 m(-2); p = 0.021) were observed, whereas in Bcl-2, the difference was not significant (77.96 vs. 71.13 ng/ml). CONCLUSIONS: At the early stages of diabetic nephropathy in type 1 diabetic patients, there seems to be a dysregulation of apoptosis, as expressed by enhanced sFas levels, leading to the speculation that the prevalence of antiapoptotic mechanisms (sFas) may promote mesangial proliferation. PMID- 17409759 TI - Solitary intracranial Castleman's disease, plasma cell variant: a case report. AB - OBJECTIVE: We report a case of plasma cell variant of Castleman's disease confined to the meninges. CLINICAL PRESENTATION AND INTERVENTION: A 53-year-old woman presented with severe headache lasting a few months, which was insidious in onset but followed a progressive course with associated vomiting, blurring of vision and diplopia. Investigations revealed a dural-based mass that was considered both radiologically and intraoperatively as a meningioma. Total surgical excision of the mass was performed. Histologically, it was a plasma cell variant of Castleman's disease with kappa light chain restriction. Laboratory investigations and whole body computerized tomography scan showed no significant abnormality. A short course of local radiotherapy was given. The patient is well 2 years after treatment. CONCLUSION: This report highlights the rarity of solitary intracranial Castleman's disease and the difficulty in radiological and intraoperative diagnosis. PMID- 17409760 TI - Isolated adrenocorticotropic hormone deficiency with transient thyroiditis inducing an adrenal crisis. AB - OBJECTIVE: It was the aim of this study to describe a patient with isolated adrenocorticotropic hormone deficiency presenting with a variety of involuntary movements who developed an adrenal crisis due to transient thyroiditis. CLINICAL PRESENTATION AND INTERVENTION: A 61-year-old man was hospitalized with a variety of involuntary movements that were suspected manifestations of metabolic encephalopathy. After admission, his general status rapidly deteriorated to a life-threatening condition that included a degree of hyponatremia. The hyponatremia and metabolic encephalopathy provided clues toward a definitive diagnosis. After corticosteroid and sodium supplementation improved the status of the patient, endocrinological examinations revealed that he suffered from isolated adrenocorticotropic hormone deficiency followed by transient thyroiditis that induced an adrenal crisis. CONCLUSION: This case emphasizes the importance of considering hypoadrenalism when encountering hyponatremia or metabolic encephalopathy of unknown etiology. PMID- 17409761 TI - Acute myocardial infarction following sildenafil intake in a nitrate-free patient without previous history of coronary artery disease. AB - OBJECTIVE: To report the occurrence of acute myocardial infarction (MI) associated with the intake of oral sildenafil (Viagra) in a nitrate-free patient without previous history of coronary artery disease. CASE PRESENTATION AND INTERVENTION: A 50-year-old manual laborer was admitted to the hospital with acute inferoposterior wall MI occurring approximately 30 min after taking oral sildenafil 50 mg. This occurred before any attempted sexual activity. Subsequent angiography showed a 70% stenotic lesion in the midsegment of the dominant circumflex artery. The Naranjo scale indicated that sildenafil was a probable cause of MI. CONCLUSION: This report shows a rare sildenafil-associated MI in a nitrate-free patient without a previous history of coronary artery disease. PMID- 17409762 TI - Role of urinary cytology in detecting human polyoma bk virus in kidney transplant recipients. A preliminary report. AB - OBJECTIVE: To report data on the evaluation of the efficacy of urine cytology in detecting BK virus (BKV). MATERIAL AND METHODS: Papanicolaou-stained cytospin preparations of randomly collected urine samples from 8 renal transplant (RTR) patients (5 males, 3 females, age: 23-63 years) who previously tested positive for BKV by PCR in urine or blood or both were studied. Urine sediment from 3 of the patients was processed for ultrastructural examination (EM). Renal biopsies before and after BKV detection were also evaluated. RESULTS: The interval between renal transplant and urine cytology ranged between 2 and 5 years. Urine from females was difficult to assess due to vaginal contamination. In 2 of the 5 urine specimens from male recipients BKV-infected decoy cells were identified. Viral particles suggestive of BKV were identified on EM in 2 of the 3 specimens studied. Kidney biopsies showed morphologic features suggestive of BKV infection in 2 cases, 1 each detected on cytology and EM, respectively. CONCLUSION: Screening for the presence of decoy cells in urine provides a simple sensitive means for the diagnosis of BKV nephropathy in RTR. PMID- 17409763 TI - Prevalence of malaria during pregnancy and antimalarial intervention in an urban secondary health care facility in Southern Nigeria. AB - OBJECTIVE: To investigate the prevalence of malaria during pregnancy and antimalarial interventions in an urban secondary health care facility. SUBJECTS AND METHODS: Of 432 pregnant women who delivered or were attending an antenatal clinic in a secondary health care facility in Benin City, Nigeria, 199 were recruited for the study. Demographic data were obtained from the pregnant women at delivery, and maternal peripheral, placental, and cord blood samples were collected for microscopy. Among the antenatal clinic attendees, a questionnaire was used to assess their antimalarial preventive measures. RESULTS: Of the 199 pregnant women, 60 (30%) reported that they had had malaria during the preceding 3 months, and a majority of them (85%) used chloroquine. Almost all reported the drug was efficacious (98%) and well tolerated (80%). Only 18 (13%), 14 (10%), and 2 (1%) of the women had positive maternal peripheral, placental, and cord blood parasitaemia, respectively. The geometric mean (+/- SD) numbers of malaria parasites per microliter were 636.06 +/- 1,450.11 in peripheral blood, 4,250.36 +/- 13,866.01 in placental blood, and 59.50 +/- 27.58 in umbilical cord blood. Only 31 (12%) and 13 (5%) of antenatal women believed in the efficacy of insecticide-treated bed nets or sulphadoxine/pyrimethamine-based intermittent preventive therapy as antimalarial preventive measures, respectively, while 23 (9%) and 31 (12%), respectively, reported they currently use them. CONCLUSIONS: Despite the common occurrence of malaria during pregnancy, there was a limited knowledge and use of the recommended antimalarial interventions by pregnant women attending the antenatal clinic of this health care facility. There is, therefore, an urgent need to increase the implementation of antimalarial interventions during pregnancy. PMID- 17409764 TI - Factor V Kuwait alias factor V R3: a rare polymorphism of uncertain functional significance. PMID- 17409766 TI - Assessment and clinical aspects of health-related quality of life in dialysis patients and patients with chronic kidney disease. AB - BACKGROUND/AIMS: It is relevant to investigate health-related quality of life (HRQOL) in dialysis and chronic kidney disease (CKD) patients in order to optimise treatment. The aim of this study was to investigate HRQOL in dialysis and CKD patients, to compare results from patients treated with hemodialysis (HD) and peritoneal dialysis (PD) and to investigate the prediction of dialysis quality control parameters (blood hemoglobin, plasma albumin, and Kt/V) and tobacco smoking in disease-specific HRQOL. METHODS: Seventy-one HD, 59 PD, and 63 CKD patients participated in the study. Dialysis quality control parameters were measured and the patients completed the questionnaire Kidney Disease Quality Of Life. RESULTS: PD patients rated Dialysis Staff Encouragement and Patient Satisfaction better than HD patients (p< or = 0.05). Dialysis patients scored significant lower than the general population in all generic HRQOL scales (p < or = 0.01), whereas CKD patients scored lower than the general population in 5 of 8 scales (p < or = 0.05). The dialysis quality parameters did not predict dialysis patients' disease specific HRQOL, but tobacco consumption was independently associated with low scores on a number of HRQOL scales. CONCLUSION: Based on the results, it is suggested to include elements of HRQOL as a supplement to standard quality control parameters. It is also suggested routinely to include information of the beneficial effects of physical activity already in the predialysis program, and to focus on smoking as a very important risk factor. PMID- 17409767 TI - Arterial stiffness alterations during hemodialysis: the role of dialysate calcium. AB - BACKGROUND/AIMS: We investigated the way dialysate calcium (dCa) level can influence arterial stiffness (AS), measured by stiffness index (SI), a surrogate of pulse wave velocity, and reflection index (RI), a measure of the amount of pulse wave reflection, derived by digital volume pulse (DVP). METHODS: Fourteen hemodialysis (HD) patients underwent two consecutive midweek 4-hour HD treatments in randomized order with a low dCa concentration of 1.25 mmol/l (LdCa) and a high dCa concentration of 1.75 mmol/l (HdCa), respectively. Before HD and at 1-hour intervals during the subsequent 4-hour HD sessions, SI and RI measurements were obtained from DVP contour analysis. Blood pressure (BP) and heart rate (HR) were measured after each measurement of AS. Ionized serum calcium (iCa) was measured before HD and at 120 and 240 min into the HD session. RESULTS: iCa increased and decreased by 15.3 and 5.4% in the HdCa and LdCa groups, respectively, at the end of HD. SI and RI increased by 5.7 and 6% in the HdCa group, respectively, whereas they remained unchanged in the LdCa group. The treatment effect and the time x treatment effect were significant for both indices (ANOVA). BP and HR changes did not differ between treatments. CONCLUSION: Contrary to LdCa, HdCa treatment induced a BP-independent, significant increase in measured AS parameters. In the long run, HD with LdCa, by reducing the incidence of HD-induced hypercalcemia, may have a beneficial role in minimizing the cardiovascular risk related to the intermittent, intradialytic increase in AS, inherent in the chronic use of HdCa. PMID- 17409768 TI - Elevated serum sFlt-1/Ang-2 ratio in women with preeclampsia. AB - BACKGROUND: An imbalance of angiogenesis-associated factors may predispose to preeclampsia. Here, we determined the ratio of serum concentration of soluble fms like tyrosine kinase 1 (sFlt-1), a natural inhibitor of pro-angiogenic vascular endothelial growth factor (VEGF) relative to angiopoietin-2 (Ang-2), a natural antagonist of angiopoietin-1 (Ang-1) involved in promoting angiogenesis in the presence of VEGF, in women with preeclampsia. METHODS: The levels of serum sFlt-1 and Ang-2 were measured by enzyme-linked immunosorbent assay. RESULTS: Significant decrease of serum Ang-2 and the increase of sFlt-1 were observed in women with preeclampsia as compared to healthy pregnant women. The serum sFlt 1/Ang-2 ratio was strikingly increased in preeclamptic women in contrast to healthy pregnant women exhibiting lower value similar to non-pregnant women. The serum sFlt-1 concentrations tended to positively correlate with mean blood pressure (BP) in preeclamptic women, but not in healthy pregnant women. A cut-off value >0.25 in the serum sFlt-1/Ang-2 ratio showed 87.1% sensitivity and 82.8% specificity in differentiating preeclamptic women from healthy pregnant women. CONCLUSION: The serum sFlt-1/Ang-2 ratio is significantly elevated in preeclamptic women as compared to healthy pregnant women. Remarkable difference of sFlt-1/Ang-2 ratio between these two groups with excellent specificity and sensitivity suggests the clinical usefulness of the serum sFlt-1/Ang-2 ratio in diagnosing and potentially predicting the onset of preeclampsia. PMID- 17409769 TI - Long-term oral sodium bicarbonate supplementation does not improve serum albumin levels in hemodialysis patients. AB - Metabolic acidosis, a frequent event in hemodialysis patients, has been implicated as a potential cause of protein-energy malnutrition. Unfortunately, correction of metabolic acidosis by means of high bicarbonate concentration in the dialysate does not seem to lead to significant changes in nutritional parameters. The project was a single-arm, open-label, 12-month pilot study at a university-based tertiary care center aimed at evaluating whether correction of metabolic acidosis through long-term oral sodium bicarbonate supplementation improves serum albumin levels and other nutritional parameters in patients undergoing maintenance hemodialysis. Twenty highly acidotic hemodialysis patients patients were invited to consume an oral supplementation of sodium bicarbonate (1 g, thrice daily), for 12 months. Patients were followed at baseline and every month, until month 12. At each follow-up visit, dry body weight, BMI, blood pressure, presence of edema, venous bicarbonate, and serum albumin were measured. Total lymphocyte count, fasting total cholesterol and C-reactive protein were assessed every 2 months. At baseline and at 12 months, the subjective global assessment of nutritional status and the protein equivalent of nitrogen appearance normalized to actual body weight were determined. Plasma bicarbonate level rose from 18.1 +/- 2.7 to 22.1 +/- 4.5 mmol/l after 10 months (p = 0.001). Mean serum albumin levels were 3.8 +/- 0.2 mg/dl at baseline and 3.9 +/- 0.2 at the end of the study. Repeated measure ANOVA showed that there was no significant effect of bicarbonate treatment on serum albumin levels (p = 0.29), dry weight (p = 0.1), serum total cholesterol (p = 0.97), total lymphocyte count (p = 0.69), or C-reactive protein (p = 0.85). Mean subjective global assessment score was 4.53 +/- 0.37 at baseline and 4.58 +/- 0.54 at 12 months (p = 0.1). Mean nPNA (g/kg/day) was 0.86 +/- 0.05 at baseline and 0.85 +/- 0.08 at month 12. The present study demonstrates that long-term oral sodium bicarbonate at the dose of 1 gram thrice daily has no significant effect on nutritional status of HD patients. PMID- 17409770 TI - Comparison of incidence of peritonitis between peritoneal dialysis solution types. AB - BACKGROUND: Peritonitis is an important cause of morbidity and mortality in patients receiving peritoneal dialysis (PD). However, there are no data about the comparison of the incidence of peritonitis among PD solution types. The aim of the present study was to compare the incidence of peritonitis among PD solutions in PD patients treated either with Nutrineal or with Extraneal or with conventional glucose solutions. MATERIALS AND METHODS: A total of 147 patients (60 female and 87 male) who underwent PD were included in the study. Of these patients, 47 used only glucose solutions (group I), 79 used glucose solutions combined with Extraneal (group II) and 21 used glucose solutions combined with Nutrineal (group III). The laboratory values and demographics of the patients were noted. RESULTS: There was no significant difference in the frequency of peritonitis among the three groups. Peritonitis occurred in 14 of 47 patients (29.8%) in group I, in 28 of 79 patients (35.4%) in group II and in 6 of 21 patients (28.6%) in group III. Patients with serum albumin levels below 3 g/dl had a significantly higher peritonitis rate than patients with serum albumin levels above 3 g/dl (p < 0.05). CONCLUSION: We have shown that a low serum albumin level is an important risk factor for the development of peritonitis in CAPD patients. The PD solution does not appear to be a risk factor for the development of peritonitis in CAPD patients, although this question should be studied further with larger numbers. PMID- 17409771 TI - Depressive symptoms, major depressive episode and cognition in the elderly: the three-city study. AB - OBJECTIVES: The relationship between depression and dementia in the elderly has been extensively studied but the tools used to define depressed subjects are heterogeneous between studies. The objective of this study was to examine the relationship between depression and cognitive performances by using multiple measures of depressive state. METHODS: A sample of 7,869 nondemented community dwelling persons aged >or=65 years participated in the study. Depressive symptoms were evaluated by the Center for Epidemiological Study Depression Scale and the Mini-International Neuropsychiatric Interview was used to assess history of major depressive episodes (MDE). Cognitive function was evaluated by several tests including the Mini-Mental State Examination (MMSE), Trail Making Test, Isaacs Set Test and Benton Visual Retention Test. RESULTS: When studied in separate models, depressive symptoms were significantly related to lower cognitive performances for all neuropsychological tests after adjusting for potential confounders (p<0.0001), whereas subjects with current MDE had significantly lower performances in MMSE and Isaacs Set Test. When studied concomitantly, only high levels of depressive symptoms were related to lower cognitive performance, whereas existence of MDE (past or current) was no longer associated with cognitive performances. CONCLUSIONS: These data suggest that once current depressive symptoms are taken into account, major depression (past or current) is not associated with lower cognitive performances in a community-based sample of elderly people aged >or=65 years. PMID- 17409772 TI - Age and risk of stroke in atrial fibrillation: evidence for guidelines? AB - AIM: Guidelines for the clinical management of patients with atrial fibrillation suggest that treatment strategies for prescribing oral anticoagulant therapy should implicate change at age 60, 65 and 75 years. We examined if there is any threshold concerning risk of stroke by age. METHODS: We identified 141,493 subjects, aged 40-89 years, with an incident hospital diagnosis of nonvalvular atrial fibrillation or flutter and no previous or concomitant diagnosis of stroke in the Danish National Registry of Patients from January 1, 1980, to December 31, 2002. The subjects were followed in the Danish National Registry of Patients for the occurrence of an incident diagnosis of stroke of any nature and in the Danish Civil Registration System for emigration and vital status. We examined the risk of stroke by age in men and women using Cox regression models, which included age categorized in intervals, linear splines of age with cut points at age 60 and 75 years, or at age 65 and 75 years. We also analyzed age as a continuous variable in linear and polynomial regression models. RESULTS: During follow-up 15,964 incident strokes were reported to the Danish National Registry of Patients. The risk of stroke increased by increasing age at baseline. We did not find any evidence for a threshold concerning risk of stroke by age, and the best model fit was obtained in a third-order polynomial regression model. CONCLUSION: The risk of stroke increased gradually by increasing age, and we could not detect any threshold concerning risk of stroke by age. PMID- 17409773 TI - Galantamine for Alzheimer's disease and mild cognitive impairment. PMID- 17409774 TI - Memantine in the treatment of dementia. PMID- 17409775 TI - In vivo dilatation of the postnatal ductus arteriosus by atrial natriuretic peptide in the rat. AB - BACKGROUND: Alpha-human atrial natriuretic peptide (hANP) reportedly increases in premature infants with patent ductus arteriosus (PDA). OBJECTIVES: To clarify a possible hANP effect to reopen the postnatal ductus, we studied in vivo reopening of the postnatal DA by a recombinant hANP, carperitide, in rats. METHODS: Near term rat pups were incubated at 33 degrees C following caesarean section. The inner diameter of the ductus was measured with a microscope and a micrometer following rapid whole-body freezing. The DA constricted quickly after birth, and the inner diameter was 0.80 and 0.08 mm at 0 min (fetal state) and 60 min after birth. hANP concentration in the pup blood and the ductus-dilating effect of hANP were studied by subcutaneous injection of hANP at 60 min after birth, and by measurement 7, 15, 30 and 60 min later. RESULTS: The peak hANP concentration was 790 pg/ml at 7 min with 1 mg/kg, which is similar to the level seen in preterm infants with symptomatic PDA. hANP dilated the postnatal ductus dose dependently and maximally at 7 min after injection. hANP dilated the postnatal constricted ductus completely to 0.79 mm in diameter with a large dose (10 mg/kg) and to 0.55 mm with 1 mg/kg. CONCLUSIONS: hANP reopens the constricted postnatal DA dose dependently in rats. The increased hANP, accompanying premature PDA, may delay closure of the DA. PMID- 17409776 TI - Rho GTPases and their regulators in neuronal functions and development. AB - Neurons are specialized cell types which send out processes in order to communicate with other cells, which can be immediate neighbors or whose cell bodies are far distant. Neuronal morphology as in all cells is determined in large part through the regulation of the cytoskeleton. One of the key regulators of the actin cytoskeleton is the Rho family of GTPases. The Rho GTPases function as molecular switches to turn on or off downstream biochemical pathways depending on the stimuli. Their activities and their regulation are controlled by many other proteins such as the guanine nucleotide exchange factors and the GTPase activating proteins. The activities of some of the Rho family members are reported to be antagonistic to one another. In general, Rac and Cdc42 promote neurite outgrowth while RhoA stimulates retraction. The balance of these opposing activities of the different Rho GTPases is crucial for the morphology and function of the neurons. PMID- 17409777 TI - Efficacy of Tonimer gel spray on postoperative nasal care after endonasal surgery. AB - OBJECTIVE: To investigate the effect of the addition of Tonimer gel spray into nasal mucosa care on nasal mucosal findings and patient comfort in the postoperative period of endonasal surgery. MATERIAL AND METHOD: 40 patients who underwent endonasal surgery were included in the study. For the postoperative nasal care, isotonic saline was applied to both nasal cavities, and subsequently, Tonimer nasal gel spray was additionally applied to the right nasal cavity. Patients were examined on the 2nd, 7th, and 15th postoperative days. The findings of the examinations were scored with respect to crusting and the patient nasal comfort was assessed using the visual analog scale (VAS). RESULTS: The VAS values which measure patient comfort and crusting were significantly lower on the Tonimer side compared to control (p < 0.01). CONCLUSION: Tonimer and saline combination was found to be superior to saline per se in decreasing crusting and improving patient nasal comfort in the postoperative period. PMID- 17409778 TI - Effect of the thickness of the cartilage disk on the hearing results after perichondrium/cartilage island flap tympanoplasty. AB - OBJECTIVE/HYPOTHESIS: The aim of this study was to analyze the effect of the thickness of the cartilage disk on the hearing results after perichondrium/cartilage island flap tympanoplasty. Our hypothesis was that thinning the rigid thick cartilage disk to half of its thickness could increase the compliance and give better acoustic gain and hearing results to patients with a reconstructed tympanic membrane. STUDY DESIGN: A prospective before/after clinical trial was conducted between January 2003 and March 2004. METHOD: Patients with chronic suppurative otitis media (mucosal type), central perforations and intact ossicular chain were randomly divided into 2 groups: the 1st group was treated with the perichondrium/cartilage island flap technique using the full-thickness cartilage disk, while the 2nd group was treated with the same technique but after bisecting the cartilage to half of its thickness. Hearing was evaluated using a 4-frequency (500, 1,000, 2,000, 3,000 Hz) pure-tone average air-bone gap before and then 8-9 months after tympanoplasty, and the results were compared statistically. PMID- 17409779 TI - The role of high-resolution magnetic resonance in atypical and intractable benign paroxysmal positional vertigo: our preliminary experience. AB - BACKGROUND: The diagnosis of benign paroxysmal positional vertigo (BPPV) is easy when typical nystagmus is present. However, diagnostic doubts arise when faced with cases presenting atypical features of the positional paroxysmal nystagmus as well as the clinical course and disease evolution. METHODS: A morphological evaluation of inner ear structures via high-resolution magnetic resonance (HR-MR) studies has been performed in 2 patients that did not respond to traditional therapeutic manoeuvres. RESULTS: In 1 patient, a filling defect at the level of the middle portion of the right lateral semicircular canal (LSC) has been demonstrated, while in the second one, a geometric-architectural abnormality - a 'fold' in the LSC - has been observed. CONCLUSION: HR-MR study of the inner ear offers a meaningful contribution to interpreting the physiopathogenesis of atypical cases of BPPV. HR-MR of the inner ear is a safe and useful tool to investigate patients with atypical and intractable BPPV. PMID- 17409780 TI - Downregulation of Fanconi anemia genes in sporadic head and neck squamous cell carcinoma. AB - BACKGROUND/AIMS: Much of our understanding of human cancer has come from studies of the hereditary cancer predisposition syndromes. Fanconi anemia (FA) is an autosomal recessive disorder characterized by cellular hypersensitivity to DNA crosslinking agents, progressive bone marrow failure, and cancer predisposition to solid malignancies, especially head and neck squamous cell carcinoma (HNSCC). Since FA pathway-deficient cells are hypersensitive to DNA crosslinking chemotherapy agents, the presence of somatic FA gene inactivation in sporadic cancers may be of clinical interest. This study sought to determine the frequency of FA gene downregulation in sporadic HNSCC. METHODS: The expression of the FA genes FANCA, FANCB, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCJ, FANCL and FANCM in 11 HNSCC cell lines and 49 tongue carcinoma samples was studied with quantitative real-time polymerase chain reaction. RESULTS: Downregulation of at least one FA gene was observed in 3 of 11 HNSCC cell lines and 66% of tongue carcinoma samples. FANCB, FANCF, FANCJ and FANCM were most commonly affected by downregulation, whereas downregulation of FANCA, FANCE and FANCD2 was rare. CONCLUSION: Our data suggest that downregulation of FA genes is common in sporadic HNSCC. The clinical implications of this finding merit further study. . PMID- 17409781 TI - Olfactory dysfunction in nasal polyposis: correlation with computed tomography findings. AB - AIMS: This study evaluates if a computed tomography (CT) scan is useful to assess the olfactory loss in sinonasal disease, and if a preoperative CT scan has a predictive value for the long-term outcome regarding olfaction. METHODS: Thirty one patients with nasal polyposis were included. Olfactory function was assessed with the 'Sniffin' Sticks' test and subjective perception recorded with a visual analogue scale. CT scans were assessed with the Lund-Mackay system and the Damm nasal segmentation. Patients were retested after endoscopic sinus surgery in a follow-up appointment at least 1 year later. RESULTS: Disease in the upper meatus and the posterior portion of the middle meatus strongly affects olfactory function. Lund-Mackay scores were significantly correlated with preoperative olfactory test results. Preoperative subjective ratings had a significant correlation only with present disease in the anterior upper meatus. Postoperative results were significantly decreased. Their relative percentage change was correlated only with the preoperative presence of disease in the anterior upper meatus. No correlation was found between the Lund-Mackay score and the postoperative olfactory results. CONCLUSIONS: Olfactory dysfunction in nasal polyposis is strongly related to specific obstructed nasal areas. A CT scan has no predictive value for the long-term surgical outcome regarding olfaction. PMID- 17409782 TI - Distortion product otoacoustic emission findings in Behcet's disease and rheumatoid arthritis. AB - BACKGROUND AND OBJECTIVE: This study aimed to assess cochlear functions in Behcet's disease and rheumatoid arthritis (RA) using otoacoustic emission testing (OAE), which objectively assesses outer hair cells in the cochlea. METHODS: Patients with Behcet's disease (n = 16) and RA (n = 11) as well as 20 controls were tested using pure tone audiometry and transiently evoked (TEOAEs) and distortion product OAEs (DPOAEs). RESULTS: Pure tone results in the Behcet's group were not significantly different from controls (p > 0.05). Pure tone results in the RA group were significantly different than in controls at 250, 500 and 6,000 Hz (p < 0.05). TEOAEs could be obtained in all participants. DPOAE amplitudes in the RA group and controls were similar (p > 0.05). DPOAE amplitudes were significantly higher in the Behcet's group than in controls at 1 and 2 kHz (p < 0.05). CONCLUSION: A subtle deterioration in hearing can be seen at low and high frequencies in RA, which is not related to outer hair cell dysfunction as detected by DPOAE testing. In the patients with Behcet's disease who have hearing thresholds within the normal limits, outer hair cell functions seem spared with an increased activity in the apical regions of the cochlea. PMID- 17409783 TI - Risk factors for auditory neuropathy/auditory synaptopathy. AB - AIMS: It was the aim of this study to describe risk factors in auditory neuropathy/auditory synaptopathy (AN/AS). METHODS: Between 1997 and 2005, we diagnosed 37 children with AN/AS. They underwent a critical chart review for risk factors and etiological coincidences in this idiosyncratic disorder. RESULTS: Eighteen neonates had a history of prematurity and low birth weight. Hyperbilirubinaemia was present in 13 children. Three patients had evidence of infection during pregnancy, and AN/AS was associated with complex syndromal diseases in 2 cases. A congenital, familial pattern was seen in 2 siblings. Seven patients had idiopathic AN/AS. CONCLUSION: Rather than being a single etiological entity, AN/AS comprises a spectrum of risk factors and associated problems affecting the cochlea and the auditory pathway. This study shows that the majority of AN/AS in children is the result of perinatal problems and is not genetic in origin. Hyperbilirubinaemia is a common and etiologically significant finding in infants suffering from AN/AS. Thus, early hearing screening for AN/AS including transient evoked otoacoustic emissions and auditory brainstem response assessment among neonates with risk factors for AN/AS is crucial in order to better manage patients suffering from this disorder. PMID- 17409784 TI - Upper lateral cartilage inner mucoperichondrial flap technique for the repair of nasal septal perforation. AB - Treatment of nasal septal perforations which decrease quality of life is a surgical challenge with variable success rates. Numerous surgical methods have been proposed for closure of perforations. The technique described in this article, i.e. closure of the perforation with an upper lateral cartilage mucoperichondrial flap through an external septorhinoplasty approach, is an alternative surgical technique. Twenty-four patients with symptomatic nasal septal perforation were operated on using this technique and followed up for a mean duration of 28 months. Preoperative and postoperative symptom scores for crusting, nasal obstruction, bleeding, discharge, whistling, headache, nasal pain, snoring and olfactory loss as well as overall discomfort levels were compared using a visual analogue scale. Complete closure of the perforation was achieved in 19 patients. PMID- 17409785 TI - Prevalence of human papillomavirus DNA in cholesteatomas. AB - Cholesteatomas show histomorphological features like papillary growth and koilocytosis, which are characteristic of lesions induced by human papillomaviruses (HPV). Two previous studies investigating the possible role of HPV in the development of cholesteatoma had detected HPV-6 and HPV-11 DNA with a prevalence differing from 3 to 36%. The aim of the presented study was to evaluate the prevalence of different HPV types in cholesteatomas using a sensitive detection system for HPV DNA. Twenty-nine biopsies from cholesteatomas were screened for HPV DNA with a 2-step broad-spectrum PCR (PCR and nested PCR). HPV-positive products were directly sequenced by means of a cycle sequencing approach. Sensitivity of the applied broad-spectrum PCR was 0.1 copy/genome. One out of 29 biopsies showed a positive signal on the nested PCR level. Considering the low prevalence (1/29 biopsies) of detected HPV DNA in cholesteatomas, infections with common HPV types are unlikely to be a causative factor. PMID- 17409786 TI - Fred J. Epstein: a historical vignette. PMID- 17409787 TI - Real-time neuronavigation with high-quality 3D ultrasound SonoWand in pediatric neurosurgery. AB - Intraoperative ultrasound (IOUS) serves as a basic imaging tool in neurosurgery. However, its low quality and difficulty in interpreting the images make its use as a resection control tool and navigation system cumbersome. We present our experience using a high-resolution 3D IOUS system combined with a navigation system in pediatric cranial surgery. We retrospectively reviewed 16 pediatric neurosurgical procedures in which a high-resolution 3D IOUS combined with a navigation system was used. The system enables basic navigation based on preoperative computed tomography or magnetic resonance imaging scans. In addition, IOUS images serve as a data set for updated intraoperative navigation. The indications for IOUS were preoperative navigation to define the skin incision and exact craniotomy site, and for real-time neuronavigation and resection control during tumor removal. The added time per case was short and no technical difficulties were encountered. High-resolution 3D IOUS combined with navigation systems has advantages for the pediatric neurosurgical population, including both real-time basic navigation and improved resection control. PMID- 17409788 TI - Brainstem tumors: where are we today? AB - Brainstem tumors comprise 10-20% of all pediatric central nervous system tumors. The management of these tumors has evolved dramatically in the past century. Once considered uniformly fatal, it is now known that brainstem tumors have distinguishing characteristics and do not behave identically. The focality and location of the lesion is determined from the clinical history, presentation, and associated imaging. Based on these findings, it is possible to predict the behavior of the tumor and choose an appropriate intervention. Focal lesions have a good prognosis and are treated operatively while diffuse lesions have a poor prognosis and are managed medically. This article reviews the current classification of brainstem tumors, current management options and future directions for the treatment of these rare tumors. PMID- 17409789 TI - Corpus callosotomy for treatment of pediatric epilepsy in the modern era. AB - OBJECTIVE: The purpose of this study was to evaluate seizure outcome in children with intractable secondary generalized epilepsy without a resectable focus who underwent complete corpus callosotomy and compare these results to those of anterior two-third callosotomy. METHOD: Data were obtained for all patients who underwent a corpus callosotomy from 2000 to 2005. The study involved 37 patients. Eleven patients had anterior two-third corpus callosotomy compared with 28 patients who underwent complete corpus callosotomy. Two of these patients had completion of their callosotomy following initial partial callosotomy. Seizure type, seizure frequency, and family satisfaction were evaluated for all patients pre- and postoperatively. RESULTS: A reduction of >or=75% in seizures occurred in 75% of the total-callosotomy patients compared to 55% of the partial-callosotomy patients. Family satisfaction for complete and partial callosotomy was 89 and 73%, respectively. No prolonged neurologic deficits were observed in either group. CONCLUSION: Complete corpus callosotomy is the most effective treatment for secondary generalized intractable seizures not amenable to focal resection in children. PMID- 17409790 TI - Endoscopic fenestration of middle fossa arachnoid cysts: a technical description and case series. AB - BACKGROUND: Arachnoid cysts are intra-arachnoidal cerebrospinal fluid collections most frequently seen in the middle cranial fossa. The optimal method of treatment for symptomatic arachnoid cysts remains controversial and includes cyst shunting, open craniotomy and endoscopic fenestration. All these techniques, however, have been associated with the development of postoperative subdural fluid collections. We describe a new endoscopic transcortical technique that attempts to avoid this complication. METHODS: Six patients with middle cranial fossa arachnoid cysts were treated with endoscopic fenestration at our institution between January 2002 and December 2005. Three cases were approached directly through the cyst, while the other 3 were approached by passing the endoscope through the rim of adjacent cortex. RESULTS: All six endoscopic fenestrations were successful in treating the arachnoid cysts. Among the 3 patients treated via a direct cyst entry, 2 cases developed significant subdural hygromas, 1 of which required aspiration. On the other hand, 1 of the 3 cases treated using a transcortical technique developed an insignificant postoperative extra-axial collection that resolved at 3 months without intervention. CONCLUSION: Endoscopic fenestration is an effective treatment for symptomatic arachnoid cysts. Endoscopic fenestration via a transcortical approach attempts to minimize cerebrospinal fluid drainage into the subdural space, avoiding the development of significant postoperative extra-axial collections, while promoting flow into the basal cisterns. PMID- 17409791 TI - Clinical significance of terminal syringomyelia in association with pediatric tethered cord syndrome. AB - An association between terminal syringomyelia (TS) and tethered cord syndrome (TCS) has been recognized. The clinical significance of TS and the need for concurrent treatment is not known. Thirty-four patients with TCS undergoing surgery for tethered cord release were evaluated for the incidence of TS. The clinical and radiological response to untethering with and without syrinx drainage was assessed. The group incidence of TS was 29.4%. There were no significant differences in the sex, age, underlying pathology or preoperative symptoms between the TCS group (TCS, n = 24) and the TCS group with TS (TCS + TS, n = 10). In the TCS group, 37.5% of the patients were asymptomatic preoperatively, and in the TCS + TS group 50% were asymptomatic preoperatively. After surgery, none of these asymptomatic patients developed symptoms. All of the symptomatic TCS + TS patients improved clinically, 12 of 15 symptomatic TCS patients improved, 2 patients were unchanged and 1 had worsening scoliosis. Syrinx did not develop in patients lacking it preoperatively. In patients where postoperative imaging was available, preoperative syringes improved. TS is clearly associated with TCS. Tethered cord release alone may be sufficient to improve preoperative symptoms and TS may be an associated phenomenon that does not mandate separate treatment. PMID- 17409792 TI - Neurosurgical management of intramedullary spinal cord tumors in children. AB - The majority of intramedullary spinal cord tumors in children are low-grade glial tumors. They become symptomatic with pain, neurologic deficits or spinal deformity. The diagnosis is most readily obtained using magnetic resonance imaging. The natural history is significant for slow progression of symptoms. Surgery is the best treatment and is also indicated to confirm the histological diagnosis. In case of a low-grade tumor or a vascular lesion such as hemangioblastoma or cavernoma, a total or near-total resection is attempted. For astrocytomas the resection almost always remains biologically incomplete, but a near-total resection is still associated with a long progression-free survival. Neurologic morbidity is relatively low during long-term follow-up but can be up to 30% for transient motor deficits. The risk for neurologic deterioration is higher for patients with pronounced dysfunction preoperatively. This is an important argument for early surgical resection. Surgery is performed using the spectrum of microsurgical techniques as well as advanced technology, e.g. lasers and intraoperative neurophysiological monitoring with motor evoked potentials. High-grade tumors are resected conservatively and treated with radiation and chemotherapy. The prognosis of high-grade glial tumors remains poor. PMID- 17409793 TI - Tethered cord syndrome: an updated review. AB - Tethered cord syndrome (TCS) is a diverse clinical entity characterized by symptoms and signs which are caused by excessive tension on the spinal cord. The majority of cases are related to spinal dysraphism. TCS can present in any age group, and presentations differ according to the underlying pathologic condition and age, with pain, cutaneous signs, orthopedic deformities and neurological deficits being the most common. Surgical untethering is indicated in patients with progressive or new onset symptomatology attributable to TCS. The surgical strategy aims to release the tethering structure and thus the chronic tension on the cord. Early operative intervention is associated with improved outcomes. Pain relief is accomplished in almost all cases. Realistic surgical goals include relief of pain and stabilization of neurological function, although improvement in function is often seen. Cord untethering can also halt the progression of scoliosis. The benefits of surgery are debated in asymptomatic patients and patients with normal imaging. PMID- 17409794 TI - Pure red cell aplasia and thymoma. PMID- 17409795 TI - Cytokine gene therapy for malignant pleural mesothelioma. PMID- 17409796 TI - Lung cancer in HIV-infected patients in the era of highly active antiretroviral therapy. AB - BACKGROUND: Infection with the human immunodeficiency virus (HIV) and lung cancer represent two problems beginning in the 20th century that are of epidemic proportions. By the end of the 20th century, therapeutic programs of modest efficacy had been developed for both. Because both HIV infection and lung cancer are common, it is not surprising that a number of patients would be afflicted with both diseases simultaneously. There is a very limited literature regarding the treatment and outcome of patients with both diseases, particularly since the advent of highly active antiretroviral therapy (HAART) for HIV infection. METHODS: We retrospectively reviewed our tumor registry to ascertain cases with concurrent lung cancer and HIV diagnoses since 1996, at the advent of HAART. Twenty-nine patients were identified at the University of Maryland, and five additional cases from an affiliated institution were identified. RESULTS: Thirty patients had non-small cell lung cancer, and four patients had small cell lung cancer. Of the 30 patients with non-small cell lung cancer, 27 had stage IIIb/IV disease and were analyzed for outcome on the basis of CD4 counts and HAART therapy. Patients with CD4 counts >200 or those on HAART had numerically, though not statistically, superior survival. Patients were able to receive standard chemotherapy regimens, and the overall survival was 5.2 months. CONCLUSIONS: This single-institution analysis appears to indicate that there is an increasing incidence of patients with lung cancer and HIV infection. Patients with advanced NSCLC who are HIV positive with CD4 counts >200 can be treated with chemotherapy and demonstrate survival comparable to that of patients without HIV infection. PMID- 17409797 TI - Does intensive follow-up alter outcome in patients with advanced lung cancer? AB - BACKGROUND: Despite aggressive multimodality treatment, 5-year survival of stage III non-small cell lung cancer (NSCLC) remains <30%. To detect relapse, progression, or development of a second primary cancer early, many clinicians perform follow-up scans. To assess the impact of routine scanning, we compared clinical trial patients who had study-mandated scans with those treated off-study who had less intensive radiologic follow-up. METHODS: The hospital cancer registry and trials databases were searched for patients with locally advanced NSCLC who had undergone multimodality treatment with curative intent. Baseline demographics were collected as well as frequency and results of clinical and radiologic follow-up. RESULTS: Forty trial patients and 35 nontrial control patients were identified. Trial patients underwent significantly more imaging, particularly in the first 2 years (2.9 versus 2.0 body scans per year, p = 0.0016; 1.1 versus 0.4 brain scans per year, p < 0.001) but did not have more frequent follow-up visits. Forty-five cancers were detected (41 relapses, four metachronous primary tumors) in 44 (59%) patients. Of these, 28 (64%) sought medical attention that led to detection before a scheduled appointment or procedure. There was no significant difference in time to relapse or second primary in trial and nontrial patients (p = 0.80). Twenty-three patients had localized relapse, but only 15 could be treated with curative intent. Despite the trial group demonstrating a higher number of asymptomatic cancers and being offered potentially curative therapy more frequently, there was no significant difference in survival between trial and nontrial patients. CONCLUSION: In patients with locally advanced NSCLC, frequent cross-sectional imaging does not alter survival after combined modality therapy. PMID- 17409798 TI - Prognosis of resected non-small cell lung cancer patients with intrapulmonary metastases. AB - BACKGROUND: In the current TNM staging system revised in 1997 for lung cancer, intrapulmonary metastases (PM) are classified into two categories: PM1 (in the same lobe of the primary tumor), designated as T4; and PM2 (in a different lobe), as M1. There have been no large-scale analyses on PM in non-small cell lung cancer (NSCLC) patients. We collected data nationwide in Japan for 7408 lung cancer patients undergoing surgical resection during a single year, 1994. We analyzed the long-term survival of NSCLC patients to evaluate the prognostic impact of PM in relation to other prognostic factors. METHOD: Medical records of 6525 NSCLC patients undergoing surgical resection during a single year, 1994, were analyzed as a subset work of the Japanese Joint Committee of Lung Cancer Registry. The committee sent a questionnaire on outcome and clinicopathological profiles to 303 institutions. RESULTS: There were 6080 PM0 (no PM), 317 PM1, and 128 PM2 patients. The 5-year survival rates were 55.1% for PM0 patients, 26.8% for PM1, and 22.5% for PM2 patients, respectively. The differences in survival between patients with PM0 and PM1 and between patients with PM0 and PM2 were significant (p < 0.001, respectively); the difference in survival was not significant between patients with PM1 and PM2 (p = 0.298). In R0 and N0 patients, survival differences were similar for PM0, PM1, and PM2 patients. Significant survival difference was detected between T3 and PM1 (p = 0.0317) and between PM1 patients and T4 patients excluding PM1 (p = 0.0083). The 5-year survival rates of PM2 patients and M1 patients excluding PM2 were 22.5% and 20.5%, respectively, and there was no significant difference between the groups (p = 0.434). CONCLUSION: There was no significant survival difference between NSCLC patients with PM1 and PM2. The survival of patients with PM1 was between that of the T3 and T4 patients excluding PM1. PMID- 17409799 TI - A phase II study of concurrent carboplatin and paclitaxel and thoracic radiotherapy for completely resected stage II and IIIA non-small cell lung cancer. AB - BACKGROUND: To determine the feasibility of combining concurrent carboplatin/paclitaxel and thoracic radiotherapy (TRT) for completely resected stage II and IIIA non-small cell lung cancer. METHODS: Eligibility stipulated gross total resections with involved lymph nodes (N1 or N2), pathologic stage II or IIIA non-small cell lung cancer. TRT consisted of 50.4 Gy in 28 fractions with a boost of 10.8 Gy for extranodal extension (ENE) or 16.2 Gy for involved surgical margins. Chemotherapy was administered every 3 weeks: carboplatin (area under the curve of 5) and paclitaxel (175 mg/m2) during TRT for two cycles, with doses increased to an area under the curve of 7.5 and 225 mg/m2, respectively, for two cycles after TRT. Cox multivariate regression analysis was used to confirm independent predictors of outcome among clinical and treatment-related factors: age, T stage, N stage, presence of ENE, presence of involved surgical margins, histopathology. RESULTS: Between April 1997 and March 2001, 42 patients were enrolled. Two patients were deemed ineligible due to having T4 disease, leaving 40 patients for analysis. Ninety-two percent (37/40) of patients had T1 or T2 disease; 60% (24/40) had N2 disease. Nine patients (22.5%) had ENE and 15% (six patients) had involved surgical margins. At a median follow up of 37 months (range, 3-103; median, 68 months for living patients), the 2- and 5-year Kaplan Meier estimates of local regional control, freedom from distant metastasis, freedom from brain metastasis, and overall survival were 92% and 88%, 77% and 59%, 87% and 71% and 72% and 44%, respectively. Fourteen patients developed distant metastasis as the initial site of failure, eight of whom had brain metastasis. Brain metastasis was the only site of failure in four of the eight patients. Multivariate regression analysis demonstrated that the only independent predictor of overall survival was histology (p = 0.02). Patients with adenocarcinoma had a 5-year overall survival of 28% versus 68% for all other cell types. There were no independent predictors of distant metastases or brain metastases on multivariate regression analysis. Treatment was tolerated reasonably well: 92% of patients (37/40) received the planned doses of TRT; 67% of patients (27/40) received all four cycles of chemotherapy. Five patients developed grade 3 esophagitis, and three patients experienced grade 3 pneumonitis. Two patients experienced grade 5 toxicity. One was treatment related due to a patient who developed grade 3 esophagitis who developed an aspiration pneumonia that progressed to acute respiratory distress syndrome. CONCLUSIONS: Our results support the Radiation Therapy Oncology Group 97-05 findings and suggest that with new and better tolerated chemotherapy regimens the strategy of concurrent TRT and chemotherapy after completely resected stage II and IIIA non small cell lung cancer should be further explored. PMID- 17409800 TI - Influence of apoptosis and cell cycle regulator proteins on chemotherapy response and survival in stage IIIA/IIIB NSCLC patients. AB - BACKGROUND: Prognosis for non-small cell lung cancer (NSCLC) patients is very poor. Prediction of the response to treatment in individual patients may be possible using molecular biological alterations such as clinical biomarkers. We investigated the predictive value of apoptosis and cell cycle regulator proteins for neoadjuvant chemotherapy response in stage IIIA/IIIB NSCLC patients. METHODS: We evaluated p53, bcl-2, p21WAF1/CIP1, p27Kip1, and Ki67 immunohistochemical expression and apoptotic index in mediastinal lymph node metastases from 23 IIIA and 10 IIIB NSCLC patients before treatment with neoadjuvant platinum-based chemotherapy. Univariate analysis was performed to evaluate the relationship between protein expression and survival or time to progression (TTP). RESULTS: Median follow-up was 25 months (range, 4-112), median TTP was 11 months (range, 0 112), and median overall survival was 22 months (range, 4-112). Of 32 assessable patients, 18 (56%) had stable disease, 12 (38%) had a PR, and two (6%) had progressive disease. Of the 22 patients assessable for pN2 following chemotherapy, 16 (77%) were positive. Univariate analysis showed that shorter TTP correlated with progressive disease (p = 0.000), positive pN2 after chemotherapy (p = 0.026), high Ki67 (p = 0.022), and high p21WAF1/CIP1 (p = 0.038). CONCLUSION: Our results suggest that in IIIA/IIIB NSCLC patients, a high level of p21WAF1 expression in mediastinal lymph node metastases before neoadjuvant platinum-based chemotherapy is associated with a poor outcome. Our results suggest that expression of p21WAF1, which plays a role in preventing apoptosis, may be significant when selecting chemotherapy for NSCLC patients. PMID- 17409801 TI - Phase II study of gefitinib, an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), and celecoxib, a cyclooxygenase-2 (COX-2) inhibitor, in patients with platinum refractory non-small cell lung cancer (NSCLC). AB - BACKGROUND: Gefitinib, an epidermal growth factor receptor tyrosine kinase inhibitor, has demonstrated a response rate of 9%-18% in relapsed non-small cell lung cancer (NSCLC) patients. The probability of response to gefitinib was not influenced by response to previous chemotherapy. Preclinical studies have suggested that celecoxib, a cyclooxygenase-2 inhibitor, has antitumor activity in NSCLC and can enhance the activity of EGFR inhibitors. We conducted a phase II study evaluating the combination of gefitinib and celecoxib in platinum refractory NSCLC patients, defined as patients whose disease had progressed on platinum-based chemotherapy or within 3 months of completing such therapy. METHODS: Platinum-refractory NSCLC patients with performance status of 0-2 and adequate organ function were included. Patients should not have been on a NSAID for 30 continuous days before study enrollment. Patients were treated with gefitinib 250 mg daily and celecoxib 400 mg twice daily. Disease assessment was performed every 8 weeks. RESULTS: Twenty-seven patients were enrolled. The response rate was 7% (2/27). The median time to progression was 2.2 months, and the median survival was 4.6 months. One female, nonsmoking patient is progression free more than 3 years after study enrollment. The drug combination was well tolerated, with the most common adverse effects being skin rash and diarrhea. CONCLUSION: In unselected platinum-refractory NSCLC patients, the response rate to the combination of celecoxib and gefitinib was similar to that observed with gefitinib alone. PMID- 17409802 TI - A randomized phase II trial of two schedules of docetaxel in elderly or poor performance status patients with advanced non-small cell lung cancer. AB - BACKGROUND: We conducted a multicenter randomized phase II trial to evaluate two schedules of single-agent docetaxel in the first-line treatment of elderly and performance status (PS) 2 patients with advanced non-small cell lung cancer (NSCLC). METHODS: Patients 70 years of age and older with a PS 0-1 or patients of any age and PS 2 were randomly assigned to docetaxel 75 mg/m2 on day 1 every 3 weeks or 30 mg/m2 on days 1, 8, and 15 every 28 days. The primary end point was frequency of grade 3/4 toxicities. Health-related quality of life, response, and survival were secondary end points. RESULTS: Fifty-five patients were randomized to received docetaxel every 3 weeks and 56 to receive docetaxel weekly. Hematologic toxicity, primarily grade 3/4 neutropenia, was significantly lower in the weekly schedule (0% versus 44%; p < 0.001). Health-related quality of life was similar between the two arms. Efficacy parameters were not significantly different, with a trend toward better survival in the weekly schedule group (6.7 versus 3.5 months). Patients with PS 0-1 had a significantly longer survival compared with PS 2 patients (7.8 versus 2.9 months; p < 0.001). A subset analysis of 30 octogenarian patients revealed similar outcomes as in 70- to 79-year-old patients. CONCLUSION: Weekly docetaxel is associated with less neutropenia and a trend toward improved survival in elderly or PS 2 patients. PS rather than age is the primary determinant of outcome in this population. Octogenarians benefited from weekly docetaxel. Future studies should separate elderly patients from PS 2 patients. PMID- 17409803 TI - Phase II trial with carboplatin and bendamustine in patients with extensive stage small-cell lung cancer. AB - BACKGROUND: Bendamustine is an alkylating agent with hybrid activity and proven efficacy in small-cell lung cancer associated with a favorable toxicity rate. This phase II study of carboplatin/bendamustine was conducted to evaluate the efficacy of this combination in patients with extensive disease small-cell lung cancer (ED-SCLC). METHODS: Fifty-six untreated patients with ED-SCLC were enrolled. Their median age was 63 years. Sixty-seven percent of patients were male and 18% had a World Health Organization performance status of 2. Bendamustine was administered as a 30- to 60-minute infusion at a dose of 80 mg/m2 on days 1 and 2, and carboplatin was given at an area under the curve of 5 on day 1 of a 21-day cycle. RESULTS: Fifty-five patients were assessable for response and toxicity. The overall response rate was 72.7% (95% confidence interval: 59%-84%), with one complete remission (1.8%). The median time to progression was 5.2 months (95% confidence interval: 4.2-5.6). At the time of evaluation, 71% of the patients had died. The median survival time reached 8.3 months (95% confidence interval: 6.6-9.9). The major toxicity of this regimen was myelosuppression, including grade 3 or 4 neutropenia (46%), thrombopenia (26%), anemia (15%), and infections (11%). Toxic death was recorded in two patients (3.6%). CONCLUSIONS: The carboplatin/bendamustine regimen is a well-tolerated cytostatic combination in ED-SCLC with activity comparable with that of other platinum-based regimens. Further investigations, such as a phase III trial, are currently planned. PMID- 17409804 TI - Phase II trial of pralatrexate (10-propargyl-10-deazaaminopterin, PDX) in patients with unresectable malignant pleural mesothelioma. AB - BACKGROUND: Several previous clinical trials have shown that malignant pleural mesothelioma is responsive to antifolates. The dihydrofolate reductase inhibitor, pralatrexate, has a favorable toxicity profile, primarily limited to stomatitis, and has demonstrated activity in patients with non-small cell lung cancer. In mesothelioma cell lines and xenografts, pralatrexate demonstrated significant antitumor activity. METHODS: We conducted this phase II study to determine the response rate of malignant pleural mesothelioma to pralatrexate at a dose of 135 mg/m2 i.v. every 2 weeks. After a protocol amendment, patients were supplemented with vitamin B12 and folic acid at the time of starting therapy. RESULTS: A total of 16 assessable patients were enrolled. No complete or partial responses were observed. Two patients with epithelioid histology had minor responses. Three other patients remained on study with stable disease for 9, 9, and 48 months. The median time to progression was 3 months. The overall median survival time was 7 months (95% confidence interval: 3.2-16.2 months) and the one-year survival was 31% (95% confidence interval: 15%-65%). Three patients (19%) had grade 2 stomatitis, eight (50%) had grade 3, and one (6%) had grade 4. CONCLUSIONS: With this particular dose and schedule, pralatrexate as a single agent had no activity in malignant pleural mesothelioma. PMID- 17409805 TI - The effect of gefitinib on B-RAF mutant non-small cell lung cancer and transfectants. AB - We previously reported one patient with squamous cell carcinoma of the lung that showed the long-term effect to gefitinib with complete response. In the present report, we examine the epidermal growth factor receptor (EGFR) and K-RAS, HER2, and B-RAF mutations in this patient to find a B-RAF exon11 mutation, resulting in a substitution of valine by phenylalanine at codon 470 (V470F) as a novel type of B-RAF mutation in human cancers. In addition, the fluorescence in situ hybridization analysis for EGFR showed the high polysomy status. B-RAF is a nonreceptor serine/threonine kinase whose kinase domain has a structure similar to other protein kinases, including EGFR members. Of interest, the B-RAF V470F mutation corresponds to a position similar to the EGFR G719X mutation located on the phosphate binding (P)-loop of EGFR that clamps ATP into the catalytic cleft. This observation suggests that gefitinib may have an anti-cancer effect on B-RAF mutant tumors. Indeed, previous reports demonstrated that H1666 cells harboring B RAF G465V mutations showed sensitivity to gefitinib, inhibiting phosphorylation of ERK1/2. We examined the effect of gefitinib on transient transfectants of the B-RAF mutant, but no drastic inhibition of ERK1/2 phosphorylation that was one of the downstream molecules of B-RAF was induced by gefitinib. In summary, we found a novel B-RAF V470F mutation in lung squamous cell carcinoma that showed response to gefitinib. However, our in vitro investigation did not explain the response observed in this particular patient. Further investigation is necessary to elucidate the mechanism of tumor sensitivity to EGFR tyrosine kinase inhibitors. PMID- 17409806 TI - Thymoma with paraneoplastic syndromes, Good's syndrome, and pure red cell aplasia. PMID- 17409807 TI - A translational view of the molecular pathogenesis of lung cancer. AB - Molecular genetic studies of lung cancer have revealed that clinically evident lung cancers have multiple genetic and epigenetic abnormalities, including DNA sequence alterations, copy number changes, and aberrant promoter hypermethylation. Together, these abnormalities result in the activation of oncogenes and inactivation of tumor-suppressor genes. In many cases these abnormalities can be found in premalignant lesions and in histologically normal lung bronchial epithelial cells. Findings suggest that lung cancer develops through a stepwise process from normal lung epithelial cells towards frank malignancy, which usually occurs as a result of cigarette smoking. Lung cancer has a high morbidity because it is difficult to detect early and is frequently resistant to available chemotherapy and radiotherapy. New, rationally designed early detection, chemoprevention, and therapeutic strategies based on the growing understanding of the molecular changes important to lung cancer are under investigation. For example, methylated tumor DNA sequences in sputum or blood are being investigated for early detection screening, and new treatments that specifically target molecules such as vascular endothelial growth factor and the epidermal growth factor receptor are becoming available. Meanwhile, global gene expression signatures from individual tumors are showing potential as prognostic and therapeutic indicators, such that molecular typing of individual tumors for therapy selection is not far away. Finally, the recent development of a model system of immortalized human bronchial epithelial cells, along with a paradigm shift in the conception of cancer stem cells, promises to improve the situation for patients with lung cancer. These advances highlight the translation of molecular discoveries on lung cancer pathogenesis from the laboratory to the clinic. PMID- 17409808 TI - Pulmonary blastoma with renal metastasis responds to sorafenib. AB - Biphasic pulmonary blastoma is a rare malignancy of the lung composed of proliferating epithelial and mesenchymal cells with a high vessel density at the tumor periphery. We report a rare case of renal metastasis of biphasic pulmonary blastoma that responded to sorafenib, an oral multikinase inhibitor. After 2 months of treatment with sorafenib, the renal tumor size decreased sufficiently to allow a safe laparoscopic radical nephrectomy. We believe that further studies are warranted to confirm the possible effects of sorafenib on pulmonary blastomas. PMID- 17409809 TI - Chemotherapy for relapsed small cell lung cancer: a systematic review and practice guideline. AB - PURPOSE: This clinical practice guideline, based on a systematic review, evaluates chemotherapy options for patients with relapsed small cell lung cancer (SCLC). METHODS: Relevant randomized trials and meta-analyses were identified through a systematic search of the literature. External feedback was obtained from practitioners in Ontario, and the guideline was approved by the provincial lung cancer disease site group. RESULTS: Six randomized trials met the eligibility criteria and were included for review. One randomized phase III trial of oral topotecan versus no treatment in patients receiving best supportive care found topotecan to have a significant benefit in terms of 6-month survival and quality of life. A randomized phase III trial compared outcomes of carboplatin in patients receiving a combination of etoposide and cisplatin (EP) and found no significant improvement associated with carboplatin, although it was associated with significantly higher grade 3/4 thrombocytopenia. Two randomized trials directly compared chemotherapy regimens (intravenous [i.v.] topotecan versus cyclophosphamide, doxorubicin, and vincristine (CAV); and bis-chloro ethylnitrosourea, thiotepa, vincristine, and cyclophosphamide (BTOC) versus EP), but these trials found no significant differences in terms of disease response or survival. I.v. topotecan was associated with significantly higher toxicities (grade 4 thrombocytopenia and grade 3/4 anemia) and greater improvement in patient-reported symptoms compared with CAV. Two randomized trials of topotecan treated patients comparing route of administration (i.v. versus oral) found no significant differences in terms of disease response, survival, or quality of life, although oral administration was associated with increased grade 3 or 4 diarrhea in both trials. CONCLUSION: Evidence on the clinical benefit of second line therapy in SCLC is limited. Topotecan is the most studied agent in this population; it has a response and survival benefit in comparison with placebo, but it also has greater toxicity in comparison with CAV. To date, significant differences in terms of response and survival are not evident in studied chemotherapy options. PMID- 17409811 TI - European trends in preoperative and intraoperative nodal staging: ESTS guidelines. AB - Preoperative and intraoperative lymph node (LN) staging is of paramount importance for patients with non-small cell lung cancer. The Council of the European Society of Thoracic Surgery took the initiative to organize workshops on intraoperative and preoperative mediastinal LN staging. This resulted in specific guidelines. Relevant peer-reviewed publications on these subjects, the experience of the participants, and the opinion of the European Society of Thoracic Surgery members contributing online were used to reach a consensus. For primary staging, mediastinoscopy remains the gold standard for the superior mediastinal LNs. Invasive procedures can be omitted in patients with peripheral tumors and negative mediastinal and hilar nodes on positron emission tomography scan. Positron emission tomography-positive mediastinal findings should always be cytohistologically confirmed. New minimally invasive techniques that provide cytohistological diagnosis became available. Their specificity is high, but the negative predictive value is low. If they yield negative results, an invasive surgical technique remains indicated. For restaging, invasive techniques providing cytohistological information are advisable. Systematic nodal dissection is recommended in all cases to ensure complete resection. Lobe-specific systematic nodal dissection is acceptable for peripheral squamous T1 tumors if hilar and interlobar nodes are negative on frozen section studies. The report from the pathologist should describe the number of LNs removed and studied, the overall number of metastatic LNs in each station, and the status of the LN capsule. We hope that the adherence to these guidelines will standardize and improve preoperative and intraoperative LN staging and pathologic evaluation of non-small cell lung cancer. PMID- 17409812 TI - Mediastinoscopic ultrasonography. AB - OBJECTIVES: Pretherapeutic T4 staging of centrally located lung cancer is crucial for the treatment strategy, but non-invasive imaging techniques are of low accuracy. We have developed the new imaging technique of intraoperative mediastinoscopic ultrasound (MUS) to predict technical resectability in tumors staged cT4 based on computed tomographic scanning. METHODS: Intraoperatively, a sterilizable fingertip ultrasound probe is introduced and guided through the video mediastinoscope with a modified grasper during staging mediastinoscopy. The position of the probe in front of the tracheobronchial tree and in direct contact with the vena cava and pulmonary artery reduces air interference. We reviewed the results for 24 patients with tumors staged cT4 between July 2002 and January 2006. For 18, the prediction of MUS concerning resectability could be compared with intraoperative findings at the time of thoracotomy. RESULTS: MUS visualizes all central vessels and their relation to the tumor with high accuracy. The pulmonary artery and pulmonary veins are displayed not only in their central parts but also in their interlobar branches. Of the 24 patients, 18 proceeded to thoracotomy after conclusive MUS and had tumors proved to be technically resectable in accordance with prediction by MUS. Comparison of cT (computed tomographic scan), cT (MUS) and pT revealed that T stages defined by MUS accurately predict pathologic T stages. CONCLUSION: MUS allows investigators to assess infiltration of the great vessels and the mediastinum, especially in right sided tumors. MUS will supplement endoscopic ultrasound-guided fine needle aspiration for the right upper mediastinum in staging of centrally located tumors. PMID- 17409813 TI - Remediastinoscopy. AB - Indications for remediastinoscopy include recurrent and second primary lung cancer, an inadequate first procedure, lung cancer occurring after an unrelated disease such as lymphoma, and restaging after induction therapy. Nowadays, restaging is the most frequent indication for remediastinoscopy. Only patients with proven mediastinal downstaging will benefit from a subsequent surgical resection. In contrast to imaging or functional studies, remediastinoscopy provides pathologic evidence of response after induction therapy. Although technically more challenging than a first procedure, remediastinoscopy can select patients for subsequent thoracotomy and provides prognostic information. In most recent series, sensitivity of remediastinoscopy is higher than 70% with an accuracy of approximately 85%. Survival also depends on the findings at remediastinoscopy, with patients with persisting mediastinal involvement having a poor prognosis. An alternative approach consists of the use of minimally invasive staging procedures as endobronchial or endoscopic esophageal ultrasonography to obtain initial proof of mediastinal nodal involvement. Mediastinoscopy is subsequently performed after induction therapy to evaluate response. In this way, a technically more difficult remediastinoscopy can be avoided. PMID- 17409814 TI - Video-assisted mediastinoscopic lymphadenectomy (VAMLA). AB - BACKGROUND: The development of a two-bladed spreadable videomediastinoscope in 1992 allowed increased exposure and bimanual dissection of mediastinal structures. Concurrent with technical progress in mediastinoscopy, neoadjuvant treatment of stage III lung cancer was introduced, and accuracy of pretreatment mediastinal staging became a topic at issue. In this setting, development of a videomediastinoscopic technique for complete mediastinal lymphadenectomy was the obvious thing to do. METHODS: Video-assisted mediastinoscopic lymphadenectomy (VAMLA) dissection is guided by anatomical landmarks, very similar to open lymphadenectomy. It includes en bloc resection of the right and central compartments and dissection and lymphadenectomy of the left-sided compartments. In a preliminary case-control study of 40 patients, VAMLA technique was standardized and evaluated against open lymphadenectomy. A second study investigated 130 patients with resectable lung cancer and radiographically normal mediastinum who underwent VAMLA and consecutive lung resection with mediastinal reexploration. RESULTS: VAMLA harvested significantly more nodes than open lymphadenectomy. With a mean duration of 54 minutes and a complication rate of 4.6%, VAMLA appeared applicable to clinical routine. We noted a sensitivity of 93.8%, a specificity of 100%, and a false-negative rate of 0.9%. CONCLUSIONS: In our experience, VAMLA is a feasible method of mediastinal staging. Its accuracy and radicality can equal open lymphadenectomy. However, VAMLA is minimally invasive and therefore pretherapeutically available. Its advantages might be of interest with neoadjuvant strategies, trials, involved field radiation, video assisted thoroscopic lobectomy, and left-sided tumors. PMID- 17409815 TI - Transcervical extended mediastinal lymphadenectomy: results of staging in two hundred fifty-six patients with non-small cell lung cancer. AB - Transcervical extended mediastinal lymphadenectomy (TEMLA) is a new surgical procedure for staging of the mediastinal lymph nodes in patients with non-small cell lung cancer (NSCLC). The operation is performed through the 5- to 8-cm collar incision in the neck and enables complete removal of all mediastinal nodal stations except for the pulmonary ligament nodes (station 9) and the most distal left paratracheal nodes (station 4L). Generally, TEMLA is an open procedure performed partly with mediastinoscopy-assisted and videothoracoscopy-assisted techniques. Operative technique of TEMLA includes elevation of the sternal manubrium with a special retractor and bilateral visualization of the laryngeal recurrent and vagus nerves. From January 1, 2004 to November 15, 2005, TEMLA was performed in 256 patients. Length of operation was 80 to 330 minutes (mean, 161 minutes). Complication of TEMLA occurred in 11.3% of patients with temporary laryngeal nerve palsy in 6 of 256 patients (2.3%) and permanent nerve palsy in 2 of 256 patients (0.8%). The number of dissected nodes during TEMLA was 15 to 85 (mean, 38.9). N2-3 nodes were found in 80 patients (31.3%). During subsequent thoracotomy, omitted N2 nodes were found in 5 of 138 patients, and omitted normal mediastinal nodes were found in 13 of 138 patients (9.4%). Sensitivity of TEMLA in discovery of N2-3 nodes was 94.1%, specificity was 100%, accuracy was 98%, negative predictive value was 97.2%, and positive predictive value was 100%. Preliminary results may suggest some therapeutic impact for patients who underwent TEMLA and subsequent R0 resection, with a 77.2% survival rate after the mean 24-month follow-up period. PMID- 17409816 TI - Efficacy of vinorelbine in the second-line setting and beyond in non-small cell lung cancer. PMID- 17409819 TI - Pick the winner designs in phase II cancer clinical trials. PMID- 17409820 TI - The Ras/Raf/MAPK pathway. PMID- 17409821 TI - Practitioner feedback on lung cancer practice guidelines in Ontario. AB - PURPOSE: Practitioner feedback (PF) surveys are sent to practitioners who care for lung cancer patients as each new practice guideline is completed. In this study, the PF was reviewed to assess the frequency of response to the surveys, the respondents' characteristics, the nature of the feedback, and the intention to adopt the guideline in practice. METHODS: Fourteen practice guidelines (PGs) were sent to Ontario practitioners treating lung cancer, and feedback on the PGs was obtained through either an eight- or 21-item survey. RESULTS: Between 1995 and 2002, 1198 surveys were sent to 223 practitioners. The overall response rate was 58.9% but varied by specialty (radiation and medical oncologists, 67%; thoracic surgeons, 46%; respirologists, 38%), by location of practice (cancer center, 65%; community-based practice, 55%), by geographic region of the province (highest, 72%; lowest, 42%), and by PG topic (chemotherapy, 60%; radiotherapy, 63%; combined modality therapy, 52%). The response rate to the PF surveys did not decline over time. Eighty-six percent of respondents agreed with the lung cancer guidelines and indicated that they were likely or very likely to use the PGs in their practice. CONCLUSION: The results suggest that practitioners view the guideline development process as credible and useful to guide practice. Whether the stated intention to use the guidelines will actually translate into practice requires further study. PMID- 17409822 TI - A randomized phase II trial using two different treatment schedules of gemcitabine and carboplatin in patients with advanced non-small-cell lung cancer. AB - BACKGROUND: Gemcitabine and carboplatin combination therapy is an active and tolerable regimen in the treatment of non-small-cell lung cancer (NSCLC). Twenty eight- and 21-day regimens without day-15 administration of gemcitabine are common; however, it remains unclear which offers the optimal therapeutic index. METHODS: This trial evaluated two schedules of the combination of gemcitabine and carboplatin: gemcitabine (1100 mg/m on days 1 and 8) plus carboplatin (area under the curve = 5 on day 8) every 28 days, or gemcitabine (1000 mg/m on days 1 and 8) plus carboplatin (area under the curve = 5 on day 1) every 21 days. Eligible patients in this trial had stage IIIB (with malignant pleural effusion) or stage IV NSCLC with no prior chemotherapy. The primary objective was to evaluate progression-free survival, with secondary objectives of overall survival, response rate, and toxicity. RESULTS: One hundred patients were randomized and enrolled from October of 2000 to January of 2002 into this multi-institutional study (48 for the 28-day regimen and 52 for the 21-day regimen). Baseline demographics were well matched, and a majority of patients (85%) enrolled with stage IV disease. Median progression-free survival and response rates were 3.8 months and 22.9%, respectively, with the 28-day regimen, and 4.9 months and 40.4%, respectively, with the 21-day regimen. Median survival was 8.7 months with the 28-day regimen and 8.0 months for the 21-day regimen. One- and 2-year survival rates were 34.7% and 8.7%, respectively, with the 28-day regimen, and 36.5% and 16.8%, respectively, with the 21-day regimen. Differences in progression-free survival (log-rank statistic, p = 0.5786), response rate (Fisher's exact test, p = 0.0859) and overall survival (log-rank statistic, p = 0.3568) were not statistically significant. Grade 3 to 4 hematologic toxicities occurred with a greater frequency in the 21-day regimen. No grade 3 to 4 nonhematologic toxicity (except nausea/vomiting with the 28-day regimen) was observed in more than 10% of patients in either treatment arm. CONCLUSION: Gemcitabine plus carboplatin is active and well tolerated in advanced NSCLC. Both regimens may be considered for further study. Although the 21-day regimen appeared to be associated with preferable outcomes, differences between treatment groups were not statistically significant. PMID- 17409823 TI - Pemetrexed maintenance therapy in patients with malignant pleural mesothelioma. AB - PURPOSE: To investigate the toxicity and effectiveness of pemetrexed maintenance therapy (PMT) in patients with malignant pleural mesothelioma (MPM). PATIENTS AND METHODS: Eligible were patients with histologically proven advanced MPM, WHO PS 0 2 and adequate hematological, renal and hepatic function in whom during 6 courses of pemetrexed containing induction therapy no disease progression was observed. PMT, 500 mg/m intravenously on day 1 every 3 weeks, was continued until disease progression, unacceptable toxicity, or if continuation was considered to be not in the patient's best interest. Written informed consent was obtained from all patients. RESULTS: Of the 27 patients who received induction therapy, 13 were treated with PMT. The median number of PMT courses was 4 (range = 2 to 14). No grade 4 toxicity was observed. Grade 3 neutropenia, leucopenia and anemia occurred 15%, 8% and 8%, respectively. The only non-hematological grade 3 toxicity during PMT was fatigue (15%). During PMT creatinine clearance decreased from 88 (+/-21) ml/min to 77 (+/-26) ml/min (p < 0.05). The reason to stop PMT was disease progression (69%), toxicity (23%) and in patient's best interest (8%). During PMT 23% of the patients with stable disease after induction therapy achieved a partial response. Time to progression and overall survival were 3.4 and 6.0 months versus 8.5 and 17.9 months, respectively (p < 0.0001). CONCLUSIONS: PMT in MPM patients is non-toxic, well tolerated and although promising effects on TTP and OS are demonstrated, the effectiveness of PMT should be further explored in a prospective randomized clinical trial. PMID- 17409824 TI - A literature review of molecular markers predictive of clinical response to cytotoxic chemotherapy in patients with lung cancer. AB - BACKGROUND: To find candidate genes for a predictive chemosensitivity test in patients with lung cancer by using a literature review. METHODS: Using MEDLINE searches, "in vitro chemosensitivity associated genes" and articles on association of the gene alteration with clinical chemosensitivity in lung cancer patients were selected. We calculated odds ratios (ORs) and their 95% confidence intervals (95% CIs) of response rates for patients who had tumors with or without gene alteration. Combined ORs and 95% CIs were estimated using the DerSimonian Laird method. RESULTS: Of the 80 in vitro chemosensitivity-associated genes identified, 13 genes were evaluated for association with clinical chemosensitivity in 27 studies. The median (range) number of patients in each study was 50 (range, 28-108). The response rates of lung cancer with high and low P-glycoprotein expression were 0% and 73% to 85%, respectively (p < 0.001). Glutathione S-transferase pi expression (OR 0.22, 95% CI 0.06-0.79), excision repair cross-complementing 1 alterations (combined OR 0.53, 95% CI 0.28-1.01; p = 0.055), and tumor suppressor p53 mutation (combined OR 0.25, 95% CI 0.12-0.52) were associated with clinical chemosensitivity. CONCLUSION: In total, 80 in vitro chemosensitivity-associated genes were identified in the literature, and high and low P-glycoprotein, glutathione S-transferase pi expression, excision repair cross-complementing 1 alterations, and tumor suppressor p53 mutation were candidates for future clinical trials of chemosensitivity tests in lung cancer patients. PMID- 17409825 TI - Phase I study of Paclitaxel given by seven-week continuous infusion concurrent with radiation therapy for locally advanced non-small cell lung cancer. AB - BACKGROUND: Paclitaxel is active in non-small-cell lung cancer (NSCLC) and is a radiosensitizer with a dose-response relationship that depends more on duration of exposure than peak concentration. A continuous infusion prolongs exposure and may maximize the drug-radiation interaction. The goal of this National Cancer Institute-sponsored phase I study was to determine the feasibility and toxicity of a continuous infusion paclitaxel (24 hours/day, 7 days/week, 7 weeks total) concurrent with standard radiation therapy (RT) for locally advanced NSCLC. METHODS: Eligible patients had locally advanced (T4, N1-3, M0 or Tany, N2-3, M0) NSCLC, performance status less than or equal to 2, and adequate hematological, hepatic, renal, and pulmonary function. RT was given to a total dose of 64.8 Gy at 1.8 Gy/day. Paclitaxel was delivered by infusion beginning 48 hours before and then continuously throughout the 7 weeks of RT. The paclitaxel concentration was escalated in sequential dose cohorts ranging from 0.5 to 17 mg/m/d, and each contained at least three patients in a standard phase I design. RESULTS: Twenty nine patients were enrolled. Significant grade 3+ toxicity was observed in one patient, who experienced grade 3 pneumonitis at the 6.5-mg/m/day dose level. This cohort was expanded, but none of four additional patients experienced significant toxicity. Three patients completed the 15-mg/m/day dose level without serious or dose-limiting toxicity. The two patients entered at the 17-mg/m/day dose level had grade 4 neutropenia requiring a delay in therapy of more than 1 week. The median survival of all patients was 12 months; however, 4 of 27 patients (15%) survived longer than 60 months (mean 63.4 months). CONCLUSION: The maximally tolerated and recommended phase II paclitaxel dose delivered by protracted continuous infusion is 15 mg/m/day when combined with thoracic RT. This schedule allows for the delivery of more total paclitaxel than other published regimens and may have less esophagitis than weekly paclitaxel regimens. This regimen has the potential to achieve a radiosensitizing serum concentration of paclitaxel continuously for 7 weeks without exceeding levels associated with neutropenia or neurotoxicity. There were four long-term survivors in this phase I study. These data suggest that continuous paclitaxel infusion with concurrent RT is safe and should be of interest to explore in combination with other cytotoxic or targeted therapies. PMID- 17409826 TI - A novel classification of MUC1 expression is correlated with tumor differentiation and postoperative prognosis in non-small cell lung cancer. AB - BACKGROUND: MUC1 is a transmembrane mucin that plays an important role in tumor progression. Many clinical studies have suggested that the expression pattern of MUC1 core protein can be a useful prognostic marker in various malignancies, but the prognostic significance in non-small cell lung cancer (NSCLC) remains uncertain. We performed a study to assess clinical significance, especially prognostic impact, of MUC1 expression in NSCLC. METHODS: A total of 62 patients with completely resected pathologic stage I to IIIA NSCLC were retrospectively reviewed. Histologic sections cut from primary tumors were immunohistochemically stained with an anti-MUC1 monoclonal antibody (CA15-3, clone DF3), which recognizes unglycosylated epitope of MUC1 core protein. According to MUC1 expression pattern, each patient was classified into the high-grade polarized expression (HP), the low-grade polarized expression (LP), or the depolarized expression (D) group. RESULTS: Twenty-four (38.7%), 21 (33.9%), and 17 (27.4%) patients were classified into the HP group, the LP group, and the D group, respectively. HP was exclusively seen in adenocarcinoma, mostly in well differentiated adenocarcinoma. D was correlated with progressive stage and lymph node metastasis. Postoperative survival of the D group seemed to be poorer than that of the HP group for all NSCLC patients, and the difference was enhanced in adenocarcinoma patients. CONCLUSION: A novel classification of MUC1 expression pattern (HP, LP, and D) was correlated with tumor differentiation and postoperative survival in NSCLC, especially in lung adenocarcinoma. PMID- 17409827 TI - Combined analysis of molecular and clinical predictors of gefitinib activity in advanced non-small cell lung cancer: epidermal growth factor receptor mutations do not tell the whole story. AB - BACKGROUND: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors have been introduced in the standard therapy of non-small-cell lung cancer (NSCLC), but they benefit a minority of patients. The study of molecular markers may identify the subset of patients who are the most appropriate to treat with these agents. METHODS: We analyzed 43 patients with advanced NSCLC who were treated with gefitinib, an oral EGFR tyrosine kinase inhibitor, were included in analysis. We evaluated EGFR in tumor tissue by using immunohistochemistry and fluorescence in situ hybridization. We also studied downstream molecules (AKT, ERK, p38 MAPK) and their activation status and the presence of EGFR mutations in tumor tissue in exons 18-21. RESULTS: Three patients had tumors with EGFR mutations, all of which had EGFR gene amplification with a ratio of 2 or greater (p= 0.001). There was no correlation between EGFR protein expression and gene amplification. Six patients (14%) achieved an objective response and nine (21%) had stable disease; the median survival was 162 days. EGFR mutations, high levels of AKT protein expression, rash of any grade, and no history of smoking were predictive of disease control (objective response plus stable disease). Only 3 of 15 patients (20%) with disease control had an EGFR mutation. On multivariate analysis, rash and AKT were independent predictors of disease control. Patients with rash survived longer than patients without rash. CONCLUSIONS: EGFR mutation positive tumors are present in a small fraction of patients who achieve disease control with gefitinib. Other molecular markers, such as AKT, need to be further evaluated. Clinical parameters remain major determinants of gefitinib activity in NSCLC. PMID- 17409828 TI - Lung cancer screening using multi-slice thin-section computed tomography and autofluorescence bronchoscopy. AB - BACKGROUND: Thoracic computed tomography (CT) for lung cancer screening is sensitive for the detection of early peripheral lung cancer but is not sensitive for detecting central preinvasive and microinvasive cancer. Our hypothesis is that the use of a two-step strategy, using a sputum biomarker, may increase the detection rate of lung cancer by identifying individuals at highest risk. METHODS: We completed a pilot study of 561 volunteer current or former smokers 50 years of age or older, with a smoking history of more than or equal to 30 pack years. All subjects received induced sputum examination and low-dose thoracic CT scan and were offered autofluorescence bronchoscopy. RESULTS: CT detected 2408 pulmonary nodules, 80% of which were less than or equal to 4 mm in diameter. During 2-year follow-up, 95% of these nodules were stable or resolved, with only 4% showing growth at any time. A total of 28 cancers were detected in 22 subjects: 21 by CT scan and seven by autofluorescence bronchoscopy. Overall, 0.9% nodules were malignant, but growth on more than or equal to two CT scans increased the malignancy rate to 75%. The mean diameter of malignant nodules on detection was 12.8 mm (range, 3 to 36.4 mm). However, 18% of malignant nodules were less than or equal to 4 mm in diameter when first seen. CONCLUSIONS: Multi detector row CT scanners found multiple small nodules in most subjects screened, but most were stable over the 2-year follow-up. Persistent interval growth increases the probability of malignancy from less than 1% to 75%. One quarter of detected cancers were CT occult and only seen with autofluorescence bronchoscopy. Prescreening using a sputum biomarker improved the detection rate of lung cancer from 3 to 5%. PMID- 17409829 TI - Where is the heart after left-sided pneumonectomy? PMID- 17409830 TI - PET scan in lung cancer: current recommendations and innovation. AB - In the past 10 years, positron emission tomography (PET), usually with F-fluoro-2 deoxy-D-glucose (FDG), has become an important imaging modality in patients with lung cancer. FDG-PET is recommended for the diagnosis of indeterminate pulmonary nodules, for which it is significantly more accurate than computed tomography (CT) in the distinction between benign and malignant lesions. A large body of evidence convincingly demonstrates that loco-regional lymph node staging by FDG PET (in correlation with CT images) is significantly superior to CT alone, with a negative predictive value equal or even superior to mediastinoscopy. FDG-PET also improves extrathoracic staging through detection of lesions missed at conventional imaging or characterization of lesions that remain equivocal on conventional imaging. Ongoing studies now concentrate on more advanced clinical applications, such as the planning of radiotherapy, the response evaluation after the induction of therapy, the early detection of recurrence, and the use in lung cancer screening. Technical innovations, such as PET cameras with better spatial resolution, or new radiopharmaceutical probes to study applications of PET in molecular biology hold promise for future refinements in this field. PMID- 17409831 TI - PET/CT fusion scan in lung cancer: current recommendations and innovations. AB - Combined fluorodeoxyglucose-positron emission tomography (PET)/ computed tomography (CT) imaging has the potential to become the new standard imaging modality for the staging and restaging of patients with lung cancer. PET/CT is superior to PET alone, CT alone, and visual correlation of both techniques separately. In particular, it improves T3 and T4 staging and delineation of tumors associated with atelectasis. CT contrast media enhancement is probably only still needed when a substantial mediastinal tumor component is present and delineation of tumor from vascular structures is relevant. PET/CT is very accurate in detecting mediastinal nodal disease, but false-positive results are sufficiently frequent to require sampling in some positive cases. Whole-body PET/CT is the most sensitive technique for detecting extracranial metastatic disease, unexpected additional primary malignancies, and recurrence. Innovations include therapy monitoring, prognostic information, evaluation of small-cell lung cancer, its use for radiotherapy planning, and four-dimensional respiratory gating acquisition. PMID- 17409832 TI - Integration of biomarkers and imaging. AB - Imaging studies provide essential diagnostic information in the care of cancer patients. Unfortunately, radiographic findings are not always diagnostic and thus an alternative approach with biomarkers has been suggested as part of the diagnostic evaluation. This discussion focuses on integration of biomarkers with imaging in the effort to guide patient management. PMID- 17409833 TI - Role of PET-CT in the optimization of thoracic radiotherapy. AB - PET-CT is an exciting new imaging technology that simultaneously acquires detailed structural and functional imaging information. PET is having an increasing impact on the management of loco-regionally advanced non-small cell lung cancer with radiotherapy. PET combined with CT is much more accurate than CT alone in staging lung cancer and patients treated with radical radiotherapy or chemoradiotherapy have better outcomes because of superior patient selection. PET also has the potential to improve radiotherapy planning by minimizing unnecessary irradiation of normal tissues and by reducing the risk of geographic miss. PET influences treatment planning in a high proportion of cases and therefore radiotherapy dose escalation without PET may be futile. PMID- 17409834 TI - Innovations in the radiotherapy of non-small cell lung cancer. AB - BACKGROUND: This review was performed to describe recent innovations in the radiation therapy of lung cancer. METHODS: The literature was reviewed regarding recent research in the therapy of lung cancer. Emphasis was placed on newer radiation therapy (RT) techniques. RESULTS: Advances in physics and computer technology have improved radiation delivery systems. New strategies have been used, such as altered fractionation patterns, three-dimensional treatment planning, intensity-modulated RT (IMRT), tomotherapy, stereotactic RT, and heavy ion RT. New technologies will make it possible to administer higher doses more precisely, which should result in better disease control, with less toxicity. CONCLUSION: Further research will improve the outcome of patients with lung cancer by providing more effective tools for the RT of this disease. PMID- 17409835 TI - Vaccines for lung cancer. AB - Immunotherapy is based on the knowledge that the immune system can distinguish cancerous cells from normal cells. Conceptually, this is an attractive adjuvant approach because it is highly specific and can deal with disseminated disease with minimal impact on normal tissues. In this review, we focus on strategies that use host immune machinery to generate anti-tumor effects, known as active immunotherapy. Proof of principle in lung cancer is now well established in clinical trials, although no superior approach has been defined and therapeutic efficacy remains unknown. In this review, we discuss rationale, biological theory, application, and clinical implementation to date. PMID- 17409836 TI - Lung cancer late-detection, multimodality strategy. PMID- 17409837 TI - Is the way up the way forward? Radiotherapy dose escalation for non-small cell lung cancer. PMID- 17409838 TI - Targeting mTOR signaling for lung cancer therapy. PMID- 17409839 TI - Conformal high dose external radiation therapy, 80.5 Gy, alone for medically inoperable non-small cell lung cancer: a retrospective analysis. AB - BACKGROUND: Retrospective analysis of patients with medically inoperable non small cell lung cancer treated with continuous high-dose external beam radiation therapy at the Medical University of South Carolina. METHODS: We identified 35 patients with non-small cell lung cancer treated 1998-2002. None were candidates for resection for reasons including: pulmonary function (n = 23), previous cancer (n = 9), other co-morbidities (n = 2), and refusal of surgery (n = 1). Median percent predicted forced expiratory volume in 1 second was 41.5%. Median age was 71 years. Five patients had more than one primary tumor: three were concurrently treated, two were sequentially treated. Lesion sizes were <3 cm (n = 24); 3-5 cm (n = 12), and >5 cm (n = 5). Nodal stage was as follows: N0 (n = 33) and N1 (n = 2). Radiation therapy was administered once daily: median dose was 80.5 Gy/35 fx/2.3 Gy/fx. The clinical target volume was tumor plus nodes > or =1.0 cm. V20 data were available for 12 patients, with a mean value of 15.7%. RESULTS: Thirty four patients completed treatment. Median follow-up was 23.0 months. There were 26 deaths: 19 died from non-small cell lung (73%) and seven died from co-morbid illness (27%). Median survival was 24 months (95% CI, 18.0-31.9 months). Four patients were alive with disease, and five were alive disease-free at 10- and 68 month follow-ups. Of 41 lesions, local failure occurred in 15 lesions (37%) of which 3 local failure patients (9%) failed concomitantly in untreated regional lymph nodes. There were no isolated nodal recurrences. Distant progression: 10 patients (29%) of which 6 distant progression without local failure. Two patients who both had prior lobectomies experienced grade 5 toxicities. CONCLUSION: Continuous high-dose external beam radiation therapy 80.5 Gy administered in 35 fractions was tolerated. Treatment-related death was rare (6%) and isolated to patients with prior lobectomies in an extremely high-risk population. Most mortality was lung cancer-related. The dose of 80.5 Gy in 7 weeks is supported for patients with single lesions and no prior lobectomy. Local failure dominates and higher effective doses should be explored. PMID- 17409840 TI - Significance of the number of positive lymph nodes in resected non-small cell lung cancer. AB - BACKGROUND: In the current tumor, node, metastasis (TNM) classification of lung cancer, N status is defined by the anatomic extent of lymph node metastases. In this study, we evaluated the prognostic significance of the number of positive lymph nodes in resected non-small cell lung cancer. METHODS: We retrospectively studied 289 patients with non-small cell lung cancer who underwent surgery, and we compared the prognostic significance of the number of positive nodes with the pN number by using multivariate analysis. Patients were classified into four groups according to the number of positive nodes: those without nodal metastasis were n, those with one to three positive nodes were n, those with four to six were n, and those with more than seven were n. RESULTS: The 5-year survival rate was 77% in the n patients, 58% in n, 42% in n, and 6% in n, which indicates that an increased number of positive lymph nodes was associated with poor prognosis. Among the pN2 patients, the n group had a better survival rate than the n and n groups. Multivariate analysis showed that the number of positive nodes was a significant prognostic factor, equal to the currently used pN number. Hazard ratios for pN1 and pN2 with respect to pN0 were 2.13 and 3.49; and 2.07, 3.03, and 10.4 for n, n, and n with respect to n. In addition, we found that our classification could reflect the better prognoses of skip or single-station nodal metastases. CONCLUSION: The number of positive lymph nodes is a strong independent prognostic factor in non-small cell lung cancer and may add new information to the pN categories of the current TNM classification. PMID- 17409841 TI - Bortezomib plus docetaxel in advanced non-small cell lung cancer and other solid tumors: a phase I California Cancer Consortium trial. AB - BACKGROUND: This phase I study was performed to determine the dose-limiting toxicity and maximum tolerated dose (MTD) of docetaxel in combination with bortezomib in patients with advanced non-small cell lung cancer (NSCLC) or other solid tumors. METHODS: Patients were enrolled in cohorts of three over six dose levels. Each treatment cycle was 3 weeks long and consisted of one docetaxel infusion (day 1) and four bortezomib injections (days 1, 4, 8, and 11). Dose escalation and MTD determination were based on the occurrence of dose-limiting toxicities in cycle 1 only. RESULTS: A total of 36 patients were enrolled, 26 of whom had NSCLC. All patients received at least one dose of study drug at one of five dose levels. The MTD of the combined regimen was determined to be 1.0/75 mg/m bortezomib/docetaxel. The combination was generally well tolerated. Toxicities were manageable, and no additive toxicities were observed. The most common adverse events were fatigue (67% of patients), nausea (50%), diarrhea (39%), and neutropenia (39%). Two patients with NSCLC achieved a partial response, and seven (19%) patients achieved stable disease (including six patients with NSCLC). CONCLUSION: The combination of bortezomib and docetaxel was feasible and well tolerated in patients with advanced NSCLC or other solid tumors. The recommended phase II dose is bortezomib 1.0 mg/m on days 1, 4, 8, and 11 plus docetaxel 75 mg/m on day 1, cycled every 21 days. Therapeutic doses of docetaxel and bortezomib are achievable for this combination. PMID- 17409842 TI - Phase II trial of gemcitabine-carboplatin-paclitaxel as neoadjuvant chemotherapy for operable non-small cell lung cancer. AB - BACKGROUND: The aim of this single-arm phase II study was to evaluate the efficacy, feasibility, and safety of the gemcitabine-carboplatin-paclitaxel combination as neoadjuvant chemotherapy in patients with operable non-small cell lung cancer (NSCLC). METHODS: Patients with stage IB, II, or IIIA NSCLC were given three cycles of chemotherapy followed by tumor resection. Each 21-day cycle consisted of gemcitabine 1000 mg/m on days 1 and 8, carboplatin AUC 5 on day 1, and paclitaxel 175 mg/m on day 1. RESULTS: Forty-four patients were enrolled: 18.2% of patients had stage IB, 15.9% had stage II, and 65.9% had stage IIIA NSCLC. All patients received three cycles of treatment. The clinical tumor response rate was 76.2% (32 of 42 patients; 95% CI, 60.5-87.9%). Thirty-six patients had a complete tumor resection, five of whom had a complete pathological response with no viable tumor cells in the resected tumor on histological examination. Median time to progression was 13.6 months (95% CI, 8.9, >16 months), and 26 of 44 patients (59.1%) had progressed. The 1-year disease-free survival rate was 53.6% (95% CI, 38.7-68.5%), and the 1-year survival rate was 86.0% (95% CI, 75.7-96.4%). Grade 3 and 4 neutropenia each occurred in 38.6% of patients, and grade 3 infection occurred in 2.3% of patients; grade 3 and 4 thrombocytopenia occurred in 25.0% and 0% of patients, respectively. CONCLUSION: The gemcitabine-carboplatin-paclitaxel combination showed promising efficacy and seemed to be safe and feasible as neoadjuvant chemotherapy in patients with operable-stage NSCLC. PMID- 17409843 TI - A Phase II randomized study of paclitaxel plus carboplatin or cisplatin against chemo-naive inoperable non-small cell lung cancer in the elderly. AB - Paclitaxel plus carboplatin (CAR) or cisplatin (CIS) has shown activity in the treatment of advanced non-small cell lung cancer (NSCLC). Our aim was to determine whether paclitaxel plus platinum is an appropriate regimen for chemo naive NSCLC in patients aged 70 years or older. Patients were randomized into paclitaxel plus CAR or paclitaxel plus CIS treatment arms. Treatment consisted of paclitaxel 160 mg/m and carboplatin at AUC = 6 (predicted using measured clearances and the Calvert formula) IV infusion on day 1 every 3 weeks, or paclitaxel 160 mg/m and cisplatin 60 mg/m IV on day 1 every 3 weeks. In total, 81 patients were enrolled from September 2000 to February 2005, including 40 who received CAR treatment and 41 who received CIS treatment. In all, 152 cycles of CAR (median, four cycles per patient) and 172 cycles of CIS (median, four cycles per patient) were given. Each arm had one complete response and 15 partial responses to the treatment, with overall response rates of 40% and 39%, respectively. Myelosuppression was mild in both arms, and there was no statistical difference between the two arms. Alopecia (P < 0.001), peripheral neuropathy (P = 0.017), and fatigue (P < 0.001) were more severe in the CIS treatment arm than in the CAR treatment arm. Median time to disease progression was 6.6 months in the CAR arm and 6.9 months in the CIS arm. Median survival time was 10.3 months in the CAR arm and 10.5 months in the CIS arm. In conclusion, paclitaxel plus CAR or CIS treatment is feasible in elderly patients and has similar activity. However, paclitaxel plus CAR had less non-hematological toxicity than paclitaxel plus CIS. PMID- 17409844 TI - Association between anemia arising during therapy and outcomes of chemoradiation for limited small-cell lung cancer. AB - BACKGROUND: Anemia during chemoradiation is associated with poorer outcomes in various cancers. Concurrent chemoradiation (CCRT) is standard therapy for fit patients with limited small-cell lung cancer (LD-SCLC). The objective of this study was to explore the relationship between anemia and treatment outcomes in patients with LD-SCLC treated with CCRT. METHODS: Charts of all patients with LD SCLC receiving CCRT at The Ottawa Hospital Regional Cancer Centre between January 1996 and December 2002 were reviewed. Information extracted included demographics, known prognostic factors, treatment details, and hemoglobin values from diagnosis until the completion of therapy. Factors associated with outcomes were determined by Cox regression analyses. RESULTS: One hundred thirty patients were eligible for inclusion, and their median survival was 18.1 months (95% CI, 14.8-25.1 months). By univariate analysis, poorer outcome was associated with Eastern Cooperative Oncology Group performance status > or =2, supraclavicular adenopathy, and pre-radiotherapy hemoglobin <100 g/liter. Pre-radiotherapy hemoglobin <100 g/liter was associated with both an increased risk of progression (hazard ratio 1.8; P = 0.04) and death (hazard ratio 1.9; P = 0.02) by multivariate analysis. CONCLUSION: Anemia during CCRT for LD-SCLC is common and may be associated with poorer outcome. Whether this association is causative or simply prognostic is unclear, and it is not known whether correction or prevention of anemia will improve outcome. Clinical trials evaluating different target hemoglobin levels and the roles of transfusion or erythropoietin during CCRT are needed. PMID- 17409845 TI - Prognostic value of fluorodeoxyglucose positron emission tomography in non-small cell lung cancer: a review. AB - Non-small cell lung cancer (NSCLC) remains a leading cause of cancer morbidity and mortality. Adjuvant chemotherapy improves survival in resected early-stage NSCLC. However, a significant proportion of patients with early-stage lung cancer are cured by surgery alone. There are no reliable clinical or molecular markers to predict outcomes after surgery in early-stage NSCLC. Positron emission tomography with 2-[F]fluoro-2-deoxy-D-glucose (FDG-PET) improves the accuracy of staging work-up in NSCLC. The standardized uptake value, a commonly used semiquantitative measure of FDG uptake, correlates with tumor doubling time and indices of cell cycling. Therefore, FDG-PET may be a useful predictor of outcome independent of its role in tumor staging. In this review, we critically examine the published studies on the utility of FDG-PET as a prognostic tool in patients with NSCLC and provide direction for future research. PMID- 17409846 TI - The ACIN1 gene is hypermethylated in early stage lung adenocarcinoma. AB - INTRODUCTION AND HYPOTHESIS: In recent years, many studies have performed genome wide searching for differentially methylated genes in cancer. We hypothesized that characteristic aberrant hypermethylation of CpG islands of certain genes may exist in the early stages of lung adenocarcinoma and that such alterations may be useful in the detection and treatment of early lung adenocarcinoma. METHODS: A pair of immortalized cell lines originating from atypical adenomatous hyperplasia (PL16T) and from the resected end of the bronchus of the same patient (PL16B) was searched for aberrantly and differentially hypermethylated DNA fragments by a combination of the methylated CpG island amplification and suppression subtractive hybridization methods. RESULTS: From 229 clones, we selected 15 fragments that had a genomic region meeting the criteria for a CpG island. We identified a gene, apoptotic chromatin condensation inducer 1 (ACIN1), that was hypermethylated in PL16T. A higher frequency of hypermethylation at a locus at the 5': end of the DNA fragment isolated from the ACIN1 gene was found in small sized adenocarcinoma (2 cm or less) (30/37, 81%) compared with normal lung tissue (9/37, 24%, p < 0.05). Interestingly, hypermethylation of ACIN1 was detected relatively frequently in the normal counterpart of adenocarcinoma without bronchioloalveolar carcinoma (BAC) component (7/16, 44%), but was rare in the normal counterpart of adenocarcinoma with BAC component (2/21, 10%, P < 0.05). CONCLUSIONS: We found hypermethylation of the ACIN1 gene in early stage lung adenocarcinoma. The role of methylation status in the development and malignant transformation of lung adenocarcinoma requires clarification. PMID- 17409847 TI - Massive adrenal metastasis. PMID- 17409848 TI - Evolving aspects of mesothelioma carcinogenesis: SV40 and genetic predisposition. PMID- 17409849 TI - Soluble mesothelin related protein in mesothelioma. PMID- 17409850 TI - Macroscopic complete resection: the goal of primary surgery in multimodality therapy for pleural mesothelioma. PMID- 17409851 TI - Newer issues in mesothelioma chemotherapy. PMID- 17409852 TI - Adjuvant chemotherapy for resected non-small cell lung cancer. AB - Despite improved surgical techniques, relapse and mortality rates remain high among patients with resected non-small cell lung cancer. Numerous randomized controlled trials in the 1980s and 1990s were unable to demonstrate a consistent survival benefit for adjuvant chemotherapy. However, in the past 2 years, the results of three pivotal trials have unequivocally shown the benefit to adjuvant treatment in resected non-small cell lung cancer. In this review, we describe early adjuvant trials and highlight the recent landmark studies in this disease. The focus of ongoing and future research efforts is also discussed. PMID- 17409853 TI - Atypical primary epithelioid hemangioendothelioma of the heart. AB - We report an unusual case of primary cardiac epithelioid hemangioendothelioma (EHE) with atypical features, which was treated by orthoptic transplantation with a good outcome for 10 years despite recurrent pulmonary and nodal metastases. EHE is a rare vascular tumor that belongs to the group of malignant proliferations from the new World Health Organization classification of soft tissue tumors. EHE may harbor atypical features that confer a more aggressive course, albeit better than that of conventional angiosarcomas. Histological examination of the primary cardiac tumor revealed a proliferation of large epithelioid tumor cells presenting atypical features and a mitotic index of 3 mitoses per 10 high power fields. In contrast, pulmonary metastases exhibited typical features of EHE, and CD 34 and CD 31 immunostainings strongly stained cytoplasmic vascular lumen. In this report, we illustrate the potential aggressiveness of the atypical variant of EHE and suggest that transplantation might be considered as an alternative therapy in the treatment of EHE of the heart. PMID- 17409855 TI - Monitoring EGFR-mutant lung cancers by means of the blood. PMID- 17409856 TI - Staging of the brain: "brainwashed" by health economics? PMID- 17409857 TI - Semaphorins in lung cancer. PMID- 17409858 TI - Does initial staging or tumor histology better identify asymptomatic brain metastases in patients with non-small cell lung cancer? AB - BACKGROUND: To determine whether the distribution, staging features, or tumor histology of non-small cell lung cancer (NSCLC) distinguishes neurologically symptomatic from asymptomatic patients initially diagnosed with lung cancer, and to determine whether these factors may predict the presence of brain metastasis. METHODS: We performed a retrospective review of 809 patients with NSCLC and brain metastases who were treated in our institution between January 1996 and March 2003. Patients who had brain metastasis on initial staging were included. Thoracic computed tomographic scans were reviewed for lung tumor features and staging. Neurological computed tomographic or magnetic resonance image scans were assessed for distribution of brain metastases. Medical records were reviewed for comprehensive staging, tumor histology, and neurological symptoms. Fisher's exact test was used to determine any differences among tumor histology, staging, and imaging features among patients with or without neurological symptoms. RESULTS: Of the 809 patients, 181 had brain metastasis at initial staging. Among these 181 patients, 120 (66%) presented with neurological symptoms (group 1); 61 (34%) patients were asymptomatic (group 2). Patients with adenocarcinoma and large-cell carcinoma had greater odds of brain metastases than patients with squamous cell carcinoma (p = 0.001). There were 106 (58.6%) patients with adenocarcinoma, 32 (17.7%) with large cell carcinoma, and 18 (9.9%) with squamous cell carcinoma. In both groups, most lung cancers were in the right lung with upper lobe dominance. No significant difference in tumor histology or T stage was found between groups, although group 2 was more likely to have a higher N stage. Of the 181 patients with brain metastasis, 60 (33.1%) had N0 disease, 51 (28.2%) had T1 disease, and 23 (19.2%) had no other metastasis. There was no correlation between number/distribution of brain metastases and tumor histology, although patients with disease in the cerebellum or temporal lobes had a greater likelihood of neurological symptoms (odds ratio 3.7). CONCLUSION: There was no significant difference in tumor histology, staging, or distribution between symptomatic or asymptomatic patients with NSCLC with brain metastases. The odds of brain metastases were greater in those with adenocarcinoma or large-cell carcinoma. PMID- 17409859 TI - Topotecan and paclitaxel in previously treated patients with relapsed small cell lung cancer: phase II trial of the North Central Cancer Treatment Group. PMID- 17409860 TI - Phase I/II study of gemcitabine and exisulind as second-line therapy in patients with advanced non-small cell lung cancer. AB - BACKGROUND: The study was designed to evaluate the safety and efficacy of exisulind, a selective apoptotic antineoplastic drug, in combination with gemcitabine as second-line therapy in patients with progressing advanced non small cell lung cancer. METHODS: Patients whose disease progressed more than 3 months from completion of first-line chemotherapy were eligible for this phase I/II trial. Primary end points were maximally tolerated dose and time to progression. Patients in the phase I portion of the study were treated with gemcitabine (1250 mg/m) in combination with three escalated dose levels of exisulind. Treatment involved six cycles of gemcitabine and exisulind followed by exisulind maintenance. The study was subsequently expanded to phase II. RESULTS: Thirty-nine patients (15 in phase I and 24 in phase II) were treated. The regimen was well tolerated with grade 3 fatigue and grade 3 constipation being dose limiting toxicities. The maximally tolerated dose was not reached. Dose level 3 of exisulind (250 mg twice daily) in combination with gemcitabine was used for phase II. The overall response rates were 7% (phase I), 17% (phase II), and 13% (all). Median time to progression and median and 1-year survival, respectively, were 3.7 and 9.7 months and 33% (phase I); 4.3 and 9.4 months and 41% (phase II); and 4.1 and 9.4 months and 39% (all). CONCLUSION: Although the study met its primary end point of improving time to progression (more than 4.1 months in phase II), we did not observe a clear survival advantage and thus do not plan to further investigate this schedule of gemcitabine and exisulind. PMID- 17409861 TI - Phase I and pharmacokinetic study of combination chemotherapy using irinotecan and paclitaxel in patients with lung cancer. AB - The purpose of this study was to investigate the maximum tolerated doses, dose limiting toxicities, efficacy, and pharmacokinetic profiles in the combination of irinotecan and paclitaxel. Eligibility criteria included age 75 years or younger, good performance status, adequate organ function, and unresectable non-small cell or extensive disease of small cell lung cancer. Irinotecan was administered on days 1 and 8 over 90 minutes, and paclitaxel was administered on day 8 over 3 hours after 90 minutes from the end of the irinotecan infusion. Irinotecan and paclitaxel were dose-escalated from 40 and 135 mg/m and repeated every 4 weeks. The authors also administered a higher dosage with preventive granulocyte colony stimulating factor support from day 9. Thirty-one patients were assessed for toxicities and responses. Dose-limiting toxicities were neutropenia and febrile neutropenia. The dose of irinotecan 60 mg/m and paclitaxel 200 mg/m with preventive granulocyte colony-stimulating factor support was tolerable and suitable for a phase II trial. Nine of 25 (36%) patients with non-small cell and all six patients with small cell carcinoma achieved partial response. The areas under the concentration versus time curves of irinotecan and its metabolites on day 8 were significantly higher than on day 1. This combination therapy must be planned only after careful consideration of the drug-drug interaction. PMID- 17409862 TI - Clinicopathologic characteristics of the EGFR gene mutation in non-small cell lung cancer. AB - BACKGROUND: The authors sought to define clinicopathologic features associated with mutations of the epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer (NSCLC). METHODS: The authors evaluated surgically resected NSCLC tumors for EGFR (exons 18-21) and KRAS (codons 12-13) mutations and immunohistochemistry (EGFR, phosphorylated-EGFR, and HER2/Neu), and correlated results with clinical outcome and patient and disease features. After their analysis on 159 patients was completed, they selected a second cohort of Asian patients (n = 22) and compared EGFR mutation results to place of birth and immigration to the United States. RESULTS: Of 159 patients, 14 had EGFR mutations and 18 had KRAS mutations. EGFR mutations were associated with adenocarcinoma (p = 0.002), female gender (p = 0.02), never-smoking (p < 0.0001), Asian ethnicity (p = 0.005), air bronchograms (p = 0.004), and multiple wedge resections (p = 0.03). Although statistical significance was not reached, a higher incidence of synchronous primary cancers (36% versus 17%; p = 0.09) and a smaller median tumor size (11.8 cm versus 24.0 cm; p = 0.24) were seen. There was no difference in disease-free survival; however, median overall survival in patients with EGFR mutations was shorter (3.49 versus 4.29 years; p = 0.85). EGFR mutation did not correlate with immunohistochemistry. In the second cohort of 22 Asian patients, 12 (55%) had the mutation. Of interest, there was no geographic difference in incidence of EGFR mutation. Asian women with the EGFR mutation developed adenocarcinoma at an earlier age than other lung cancer patients. CONCLUSION: There is a distinct clinical profile for NSCLC patients with the EGFR mutation. However, this mutation does not alter disease-free survival and is likely attributable to an inherited susceptibility instead of an environmental effect. PMID- 17409863 TI - Treatment of elderly non-small cell lung cancer patients with three different schedules of weekly paclitaxel in combination with carboplatin: subanalysis of a randomized trial. AB - BACKGROUND: Administration of paclitaxel on a weekly schedule in combination with carboplatin is associated with a lower incidence of neuropathy and myelosuppression. The authors conducted subgroup analysis of their randomized phase II study of three different schedules of weekly paclitaxel with carboplatin to determine the efficacy of each regimen in elderly patients (aged > or = 70 years) with advanced non-small-cell lung cancer (NSCLC). METHODS: Patients with advanced NSCLC were randomized to one of three different weekly paclitaxel/carboplatin regimens. After four cycles of chemotherapy, those with objective response or stable disease were randomized to weekly paclitaxel or observation as maintenance therapy. Four hundred three patients were enrolled in the study, of whom 111 (28%) were aged 70 years or older. RESULTS: The treatment regimen of weekly paclitaxel (100 mg/m for 3 of 4 weeks) and carboplatin (area under the curve = 6 mg/ml/min once every 4 weeks) (arm 1) was associated with the best therapeutic index overall. The median survival and 1-year survival rates were 11.3 months and 50% for patients in the > or =70 years cohort versus 11.2 months and 46% for the <70 years cohort in arm 1. Efficacy results were comparable between the two groups in the other arms as well. Grade 4 neutropenia and febrile neutropenia occurred in 13.6% and 2.3% in the > or =70 years cohort compared with 4.5% and 1.1% in the <70 years cohort in arm 1. CONCLUSION: The weekly regimen of paclitaxel administered in combination with carboplatin is tolerated well by elderly NSCLC patients and has comparable efficacy with younger patients. PMID- 17409864 TI - Phase II trial of temozolomide and irinotecan as second-line treatment for advanced non-small cell lung cancer. AB - BACKGROUND: This study was performed to evaluate the tolerability and efficacy of temozolomide and irinotecan as a second-line regimen in recurrent/metastatic non small cell lung cancer (NSCLC). METHODS: Patients with recurrent/metastatic NSCLC, including those with treated brain metastases, following one prior platinum-based regimen received temozolomide 75 mg/m daily on days 1 through 15 and irinotecan 100 mg/m on days 8 and 15 every 21 days. RESULTS: The authors treated 46 patients, of whom more that 90% had a performance status of 0 or 1. Four patients (8.7%) attained partial response and 17 (37.0%) had disease stabilization as their best response. The median time to progression was 1.8 months, median overall survival was 9.8 months, and 1-year overall survival was 34%. Grade 1/2 fatigue (63%), anemia (61%), nausea (52%), and diarrhea (44%) were the most common toxicities. Grade 3/4 leukopenia and diarrhea were each observed in 9% of patients. One unexpected death occurred, possibly related to the regimen. CONCLUSION: Second-line treatment with temozolomide and irinotecan showed tolerable toxicities. The response rates, median survival times, and 1 year survival rates were comparable to other active NSCLC agents. PMID- 17409865 TI - Esophageal cancer located at the neck and upper thorax treated with concurrent chemoradiation: a single-institution experience. AB - BACKGROUND: To characterize the treatment and outcome of patients with cervical and upper thoracic esophageal cancer, the authors retrospectively reviewed the 11 year experience from The University of Texas M. D. Anderson Cancer Center. METHODS: Thirty-five patients with M0 cervical or upper thoracic esophageal cancer and treated with concurrent chemoradiotherapy were analyzed. Median radiation dose was 50.4 Gy (range, 24.5-64.8) Gy delivered with 1.8-Gy daily fractions over 5.5 weeks. Chemotherapy was 5-fluorouracil based. Response after treatment was evaluated on the basis of radiography, biopsy, or both. The survival rates were calculated by means of the Kaplan-Meier method. RESULTS: The median follow-up for the surviving patients was 39 months. The actuarial 5-year overall survival (OS), cause-specific survival, disease-free survival, local relapse-free survival, and distant metastasis-free survival rates were 18.6%, 27.6%, 22.4%, 47.7%, and 57.0%, respectively. Patients who received a radiation dose of greater than or equal to 50 Gy had a higher complete response rate than those who received less than 50 Gy (79.2% versus 27.3%; p = 0.003). On multivariate analysis, radiation dose was the only protective factor associated with the rates of OS (p = 0.006), cause-specific survival (p = 0.003), and local relapse-free survival (p = 0.001); tumor stage was the only factor associated with rate of disease-free survival (p = 0.007). CONCLUSION: Concurrent chemoradiotherapy is an effective treatment modality for patients with cervical and upper thoracic esophageal cancer. The authors' results suggest that a total radiation dose of 50 to 65 Gy with a concurrent chemotherapy regimen may improve local control and the OS rate in this rare type of esophageal cancer. PMID- 17409866 TI - EGFR mutation of tumor and serum in gefitinib-treated patients with chemotherapy naive non-small cell lung cancer. AB - BACKGROUND: The authors evaluate the efficacy and safety of gefitinib monotherapy in chemotherapy-naive patients with advanced non-small-cell lung cancer (NSCLC). A secondary endpoint is to evaluate the relationship between clinical manifestations and epidermal growth factor receptor (EGFR) mutation status. METHODS: Japanese chemotherapy-naive NSCLC patients were enrolled. They had measurable lesions, Eastern Cooperative Oncology Group performance status of 0 to 2, and adequate organ and bone marrow function. Patients received 250 mg of oral gefitinib daily. EGFR mutations in exon 18, 19, and 21 of DNA extracted from tumor and serum were analyzed by genomic polymerase chain reaction and direct sequence. RESULTS: All 30 patients were eligible for the assessment of efficacy and safety. An objective response and stable disease were observed in 10 patients (33.3%) and nine patients (30.0%), respectively. The median time to progression was 3.3 months and the median overall survival was 10.6 months. The 1-year survival rate was 43.3%. Grade 3 toxicities were observed in seven patients. EGFR mutation was observed in four of 13 (30.8%) tumors, and two of them achieved partial response. In serum samples, three of 10 patients with EGFR mutations in the serum before treatment had a response to gefitinib. EGFR mutation was observed in 10 of 27 and significantly more frequently observed in the posttreatment samples from patients with a partial response or stable disease than in those from patients with progressive disease (p = 0.006). CONCLUSIONS: Gefitinib monotherapy in chemotherapy-naive NSCLC patients was active, with acceptable toxicities. These results warrant further evaluation of gefitinib monotherapy as a first-line therapy. The EGFR mutation in serum DNA may be a biomarker for monitoring the response to gefitinib during treatment. PMID- 17409867 TI - A single lung metastasis of endometrioid adenocarcinoma synchronous with a primary bronchioloalveolar carcinoma in the same lobe. PMID- 17409868 TI - A review of first-line treatment for small-cell lung cancer. AB - Although small-cell lung cancer (SCLC) makes up a smaller proportion of all lung cancers than it did 25 years ago, it remains a common cause of cancer mortality that requires more clinical and basic research than is currently underway. Trials of newer chemotherapy variations have failed to produce a regimen that is clearly superior to the two-drug combination of etoposide and cisplatin, which remains the standard of care for both limited and extensive stage SCLC. Paradoxically, advances in this systemic disease have come from radiotherapy innovations for limited SCLC, including addition of thoracic irradiation to systemic chemotherapy, more intense thoracic irradiation, early integration of thoracic irradiation with systemic chemotherapy, and prophylactic cranial irradiation. PMID- 17409869 TI - Chemotherapy in malignant mesothelioma: what's up, doc? PMID- 17409871 TI - Chemokines in the biology of lung cancer. AB - This is a brief review of some of the mechanisms by which members of the large family of chemotactic cytokines (known as chemokines) participate in critical features of lung cancer biology. PMID- 17409872 TI - Induction chemotherapy, extrapleural pneumonectomy, and postoperative high-dose radiotherapy for locally advanced malignant pleural mesothelioma: a phase II trial. AB - INTRODUCTION: Extrapleural pneumonectomy (EPP) and adjuvant high-dose radiation therapy (RT) are associated with a median survival of 3 years in early-stage malignant pleural mesothelioma (MPM) but of less than 1 year in locally advanced disease. Although local control after EPP and RT is excellent, most patients die of distant metastases. We designed this clinical trial to test the feasibility of induction chemotherapy followed by EPP and RT in locally advanced MPM with the ultimate aim of improving survival. METHODS: Patients with MPM and stage III or IV disease were eligible. Induction therapy was four cycles of gemcitabine and cisplatin. Patients without disease progression by computed tomography underwent EPP followed by adjuvant hemithoracic RT (54 cGy). RESULTS: From January 2002 to January 2004, 21 patients (17 men, four women; median age 60 years) were entered into the study. Histology was epithelioid in 14 patients and mixed or sarcomatoid five patients. Pretreatment disease stage was III in 13 patients and IV in six patients. Nineteen patients received induction chemotherapy. Response to induction therapy was complete in zero patients, partial in five patients, stable disease in six patients, and progression of disease in eight patients. Eight of nine patients undergoing surgical exploration had EPP. The median survival of all patients was 19 months. Patients who had an EPP had a median survival of 33.5 months. Patients with unresectable tumors had a median survival of 9 months (p = 0.01). CONCLUSION: Induction chemotherapy with gemcitabine and cisplatin followed by EPP and adjuvant RT for locally advanced MPM is feasible and leads to a better median overall survival than that previously reported with EPP and RT alone. PMID- 17409873 TI - The value of HRCT and Tc-depreotide in the evaluation of pulmonary lesions. PMID- 17409874 TI - Lack of association between sputum atypia and chronic obstructive pulmonary disease mortality. AB - HYPOTHESIS: Chronic obstructive pulmonary disease (COPD) and lung cancer are thought to share common elements in pathogenesis. The authors hypothesized that sputum atypia would reflect the processes leading to progressive airflow obstruction and might be a novel biomarker of more rapidly progressive COPD. METHODS: The authors analyzed the association between COPD death and sputum cytologic atypia in an ongoing cohort of 2013 smokers with varying degrees of airflow obstruction during the period between January 1, 1993, and July 1, 2001. RESULTS: There were 326 deaths attributed to COPD over 4495 person-years, giving a COPD death rate of 7.25 deaths per 100 person-years, which is highly elevated compared with fewer than 0.2 COPD deaths per 100 person-years for the United States population aged between 65 and 74 years. Sputum atypia was not associated with either the degree of airflow obstruction or death from COPD. COPD death was associated with age and degree of airflow obstruction, as expected. CONCLUSION: Sputum cytologic atypia is not predictive of death from COPD. As sputum cytologic grades of moderate or worse atypia are associated with a significant increase in the risk for lung cancer and do not denote a group with increased competing death rates from COPD, patients with sputum atypia are a good high risk group in whom chemoprevention and early detection studies can be conducted. PMID- 17409875 TI - Neoadjuvant chemotherapy for stage III and IVA thymomas: a single-institution experience with a long follow-up. AB - BACKGROUND: Since 1989 we have enrolled patients with clinical-radiological stage III-IVA thymomas, independent of the surgeon's judgment of resectability, into a prospective study of neoadjuvant chemotherapy-surgery and postoperative radiotherapy. In this article, we review our long-term experience of neoadjuvant chemotherapy of advanced stage (III-IVA) thymomas. METHODS: From 1989 to 2004, 30 patients with Masaoka stage III and IVA thymomas underwent neoadjuvant chemotherapy, surgery, and postoperative radiotherapy. The neoadjuvant and adjuvant chemotherapy consisted of three courses of cisplatin, epidoxorubicin, and etoposide every 3 weeks. Adjuvant radiotherapy consisted of 45 Gy for complete resections or 60 Gy for incomplete resections. RESULTS: The preoperative diagnosis of invasive thymomas was obtained for 16 patients: five by mediastinotomy, seven by video-assisted thoracic surgery, and four by fine needle aspiration. For 14 patients, no histological diagnosis was available, but a thymus-related syndrome was present in all.Twenty-seven patients are still alive (25 disease-free) and three have died (one disease-free). The 10-year survival rates were 85.7% and 76.1% for stage III and IVA thymomas, respectively (difference not significant). Only the World Health Organization pathological diagnosis significantly affected the survival, with type B3 having a worse prognosis than type AB, B1, and B2 thymomas (p = 0.02). CONCLUSION: The multimodality treatment of stage III and IVA thymomas by means of neoadjuvant chemotherapy provides good long-term outcomes in both stages of the disease. PMID- 17409876 TI - Improved diagnostic efficacy by rapid cytology test in fluoroscopy-guided bronchoscopy. AB - BACKGROUND: Fluoroscopy-guided bronchoscopy is a safe and routine method used to obtain a histologic or cytologic specimen of peripheral lung nodules, but it has low sensitivity in diagnosing malignant tumors. Although feedback from rapid cytology tests are expected to improve diagnostic rates, the value of the routine use of rapid cytology tests has not been established. MATERIALS AND METHODS: We prospectively studied 657 patients with suspected peripheral malignant lung lesions on chest computed tomography who underwent fluoroscopy-guided bronchoscopy between January 2002 and December 2004. Rapid on-site cytopathologic examinations (ROSE) were performed during bronchoscopic examinations. The additional approach to the lesions was performed immediately after conventional bronchoscopic examinations when ROSE was not considered diagnostic. RESULTS: There were 528 patients diagnosed as having malignant lesions. In 477 of these patients (90.3%), final malignant diagnosis was established by the initial bronchoscopy. Among these, 84 patients (15.9%) were diagnosed only with the additional feedback from ROSE. Of 240 peripheral lesions < or =2 cm, 174 were found to be malignant. Without ROSE, 110 (63.2%) of peripheral malignant lesions were diagnosed by bronchoscopy. The integration of ROSE enabled us to diagnose an additional 40 patients (23.0%) by bronchoscopy. ROSE improved diagnostic yield independent of the site and histology of the lesions and experience of the operators. CONCLUSION: ROSE increased the diagnostic yield of bronchoscopy from 74.4% to 90.3% and therefore is an effective reinforcement in bronchoscopic diagnosis of peripheral pulmonary malignancies. The use of ROSE in routine bronchoscopy should be encouraged. PMID- 17409877 TI - Rigid bronchoscopic intervention in patients with respiratory failure caused by malignant central airway obstruction. AB - INTRODUCTION: Bronchoscopic intervention in patients with malignant central airway obstruction provides initial palliation and stabilization of the airway, allowing the possibility of other effective therapeutic modalities, such as surgery, radiation, or chemotherapy. In critically ill patients, however, the only studies of bronchoscopic intervention are relatively small series. METHODS: To describe the advantages and limitations of rigid bronchoscopic intervention in critically ill patients, we reviewed the medical records of 36 patients (26 men; median age, 62 years; range, 29 to 76 years) who underwent emergency airway intervention for malignant central airway obstruction. RESULTS: Dyspnea was relieved in 34 of 36 patients (94.4%). After the airway was widened, additional definitive therapeutic modalities were used for 21 of 34 patients (61.8%). Patients who underwent additional definitive therapy after bronchoscopic intervention survived longer (median, 38.2 months; range 1.7 to 57.0 months) than those who did not (median, 6.2 months; range, 0.1 to 33.7 months; p < 0.001). CONCLUSIONS: These data show that rigid bronchoscopic intervention in critically ill patients with malignant central airway obstruction may be temporarily life saving and, in some patients, may serve as a "bridge" to allow time for additional therapies for longer survival. PMID- 17409878 TI - The role of transbronchial fine needle aspiration in an integrated care pathway for the assessment of patients with suspected lung cancer. AB - Transbronchial fine needle aspiration (TBNA) is a simple technique for sampling mediastinal lymph nodes and may provide additional information in patients with suspected lung cancer. However, the technique is still under-utilized, and the objective of this study was to evaluate the value of TBNA as part of an integrated pathway for the assessment of patients with suspected lung cancer. All patients referred to the lung cancer services of our institutions were prospectively evaluated. TBNA was performed in all patients with evidence of mediastinal lymphadenopathy. TBNA of one or more lymph node sites were performed in 129 of these patients. TBNA was the sole diagnostic modality in 23% of patients and provided positive staging information for 49% of patients, with adequate sampling in 71% of patients. Among patients with mediastinal adenopathy, the number of patients who required a TBNA performed to diagnose one patient with malignancy in patients suspected with lung cancer (number needed to diagnose) was 1.47 (95% confidence interval, 1.47-1.76). No complications were observed in patients who underwent TBNA. TBNA improves the diagnostic yield and staging of patients with lung cancer. Moreover, it is a simple, low-cost, and safe test, which should be incorporated into the diagnostic pathway of patients with suspected lung cancer. PMID- 17409879 TI - Weekly paclitaxel combined with monthly carboplatin in elderly patients with advanced non-small cell lung cancer: a multicenter phase II study. AB - INTRODUCTION: We designed this phase II trial to evaluate the efficacy and safety of weekly paclitaxel in combination with monthly carboplatin as first-line treatment in elderly patients with advanced non-small cell lung cancer (NSCLC). METHODS: Main eligibility criteria were histologically or cytologically proven stage IIIB or IV NSCLC, age > or =70 years, Eastern Cooperative Oncology Group performance status 0-2, and measurable disease. The 4-week-based chemotherapy regimen consisted of carboplatin infusion (area under the concentration-time curve 6 mg/ml/min) on day 1 and paclitaxel 90 mg/m as a 1-hour infusion on days 1, 8, and 15. Tumor response was evaluated using Response Evaluation Criteria in Solid Tumors criteria, and symptoms were evaluated using the Lung Cancer Symptoms Scale. Analyses were performed on an intention-to-treat basis. RESULTS: From February 2002 to August 2003, 51 patients (median age, 74 years) participated in the study. One complete and 21 partial responses were reported by the independent review committee, leading to an intention-to-treat objective response rate of 43% (95% confidence interval, 30-57%). The median progression-free and overall survivals were 7.5 (95% confidence interval, 6.2-9.4) and 13.6 (95% confidence interval, 7.5-17) months, respectively. Longitudinal evaluation of the Lung Cancer Symptoms Scale demonstrated lack of quality of life modification during the treatment period. Neurotoxicity was mild to moderate, with 6% of patients suffering from a grade 3 or 4 neuropathy. Myelosuppression was the main toxicity; 39% of patients experienced grade 3 or 4 neutropenia, 18% experienced grade 3 anemia, and 8% experienced grade 3 or 4 thrombocytopenia. There was no treatment related death. CONCLUSIONS: The combination of weekly paclitaxel 90 mg/m administered on days 1, 8, and 15 plus monthly carboplatin area under the curve 6 on day 1 of a 4-week cycle was feasible and active as a first-line treatment for elderly patients with NSCLC with a good safety profile. These results deserve further analysis to compare the standard care for these patients (monotherapies) with this doublet. PMID- 17409880 TI - Causes and outcomes of spontaneous pneumothoraces in solid tumor cancer patients: an update for the medical oncologist. AB - PURPOSE: Defined as lung collapse in the absence of a recent invasive thoracic procedure, a spontaneous pneumothorax can be a catastrophic event, leading to abrupt shortness of breath, chest pain, hypotension, and occasionally death. A dearth of present day information on this entity in solid tumor cancer patients prompted this single-institution retrospective study on current causes and outcomes. METHODS: All patients with diagnoses of "spontaneous pneumothorax" and "cancer" between 1990 and 2004 had their records retrieved and reviewed. Among 546 patients with a diagnosis of spontaneous pneumothorax, only 25 (5%) met predefined inclusion criteria that included an antecedent diagnosis of an invasive solid tumor malignancy. Lung (n = 5) and bladder cancer (n = 4) were the most common malignancies; eight patients had received radiation and one had received carmustine. Of note, 78% were smokers, 13 had chronic obstructive pulmonary disease, and 12 had no known active cancer at the time of the pneumothorax. RESULTS: Pneumothorax management was associated with great morbidity, including hospitalization in 24 patients and chest tube placement and/or surgery in most patients. Median survival for the group as a whole was 31 months, but patients with known active cancer tended to do poorly, with only a 3 month median survival. CONCLUSION: A spontaneous pneumothorax is rare, and patients with known active cancer tend to do poorly. However, even patients with no known active cancer are at risk, perhaps in part from smoking. The fact that patients with no known active cancer can live for years after this event suggests that the pneumothorax should not be assumed to be related to cancer recurrence, that cancer restaging is not always mandatory, and that there is justification for managing the pneumothorax in this subgroup aggressively. PMID- 17409881 TI - Bronchioloalveolar carcinoma is not associated with younger age at diagnosis: an analysis of the SEER database. AB - INTRODUCTION: Bronchioloalveolar carcinoma (BAC) is a unique subtype of non-small cell lung cancer (NSCLC) that is associated with female gender, Asian ethnicity, and never-smoking status. Although BAC is commonly reported to occur more frequently in young people with lung cancer, there is a lack of evidence to support this association. METHODS: We analyzed the association between age at diagnosis and NSCLC histology among 293,417 incident cases of NSCLC in the Surveillance Epidemiology, and End Results (SEER) database during the years 1973 to 2002. RESULTS: The mean age of patients with BAC (66.99 years) was similar to the mean age of all patients with NSCLC (66.44 years). The proportion of patients younger than 50 years of age was significantly smaller among patients with BAC than the overall cohort (6.06% compared with 6.90%). Although a greater percentage of women and Asian patients with lung cancer were younger than 50 years old, the proportion of patients with BAC was similar to the proportion of men and non-Asians with BAC. Finally, the prevalence of BAC histology among patients younger than 50 years did not change significantly after revision of the 1999 World Health Organization pathologic criteria for the diagnosis of BAC (risk ratio 0.93 versus 0.87, p = 0.31). CONCLUSION: BAC is not associated with a younger mean age at diagnosis, nor is it associated with an age of less than 50 years at diagnosis. Patients with mixed BAC probably have similar age characteristics compared with patients with pure BAC. PMID- 17409882 TI - The bronchioloalveolar carcinoma and peripheral adenocarcinoma spectrum of diseases. AB - Bronchioloalveolar carcinoma (BAC) develops from terminal bronchiolar and acinar epithelia, growing along alveolar septa but without evidence of vascular or pleural involvement. A final diagnosis of BAC can only be achieved from a surgical specimen. Problematically, BAC may exhibit multifocal involvement by means of diffuse aerogenous metastatic spread, making this definition inapplicable for patients with stage IIIB to IV disease from whom only small size biopsy or cytological specimens are obtained. The recent interest and potential importance of BAC and the related peripheral adenocarcinoma (ADC), mixed subtype, is attributable to mounting evidence that some, perhaps many, of what are called peripheral ADCs have arisen from and often contain BAC. BAC, in turn, appears to arise from smaller peripheral nodules, called atypical adenomatous hyperplasia. These developments could account for part of the increase in ADCs noted in some countries, in particular, in East Asia. Interest also stems from the observation that advanced ADC, often with BAC features, are responding in surprising fashion to tyrosine kinase inhibitors. Furthermore, some of the more rapid, dramatic, and durable responses occur when specific mutations in the epidermal growth factor receptor are present. Clinical characteristics often differ from other types of non-small cell lung cancers. These include frequent female occurrence, especially in East Asians; no or less smoking history; an often indolent course; distinctive chest computed tomographic findings; frequent presentation as an asymptomatic, sometimes small, peripheral nodule(s)/mass; multifocal/synchronous primary tumors; and less frequently as pneumonic-type consolidation or diffuse, inoperable lesions, the latter two often with bronchorrhea, and with chest-only disease. Relapses also are predominantly pulmonary, perhaps related to aerogenous spread, and responsible for mortality. Lobectomy is the treatment of choice for cure, even with pneumonic consolidation, but lesser procedures such as wedge resection or segmentectomy may be considered for what might be multifocal, synchronous primary tumors and for pulmonary relapses. Because of frequent lung only recurrences, lung transplantation, although performed rarely, may hold promise. PMID- 17409883 TI - Metastatic lung cancer to the pancreas. PMID- 17409884 TI - Successful real-time endobronchial ultrasound-guided transbronchial needle aspiration of a hilar lung mass obtained by traversing the pulmonary artery. AB - The utility and safety of transbronchial needle aspiration is well described. Serious complications from transbronchial needle aspiration are exceedingly rare. The addition of endoscopic ultrasonographic techniques in the form of endoscopic ultrasonography and, more recently, endobronchial ultrasonography has added valuable information regarding lymphatic anatomy of the hilum and mediastinum. In addition, the use of real-time ultrasound gives the operator visual feedback of needle placement and proximity to major surrounding structures such as the heart and great vessels. Here, the authors present the case of a 74-year-old man with a left hilar mass who underwent biopsy by means of intentional traverse of the pulmonary artery. PMID- 17409885 TI - Gemcitabine and carboplatin chemotherapy safely administered to a patient with homocystinuria. PMID- 17409886 TI - Use of the epidermal growth factor receptor inhibitors gefitinib and erlotinib in the treatment of non-small cell lung cancer: a systematic review. AB - INTRODUCTION: Inhibition of the epidermal growth factor receptor is a promising therapy in the treatment of non-small cell lung cancer (NSCLC). In this systematic review, we evaluated the role of the epidermal growth factor receptor inhibitors gefitinib and erlotinib in the treatment of patients with advanced NSCLC. METHODS: Relevant randomized trials published as articles or abstracts were identified through a systematic search of the literature from 1975 to November 2005 by two independent reviewers. RESULTS: Twelve randomized trials met the predefined eligibility criteria for this systematic review. Four large placebo-controlled trials demonstrated that the addition of gefitinib or erlotinib to platinum-based first-line chemotherapy did not significantly improve overall survival or time-to-disease progression. A large placebo-controlled trial revealed a clinically and statistically significant survival benefit for erlotinib therapy as second- or third-line systemic therapy. The results of a single placebo-controlled trial and two phase II trials suggest that modest tumor response rates and symptom control can be achieved with gefitinib as second-line or subsequent therapy; however, a statistically significant survival benefit was not found for gefitinib compared with placebo. CONCLUSION: There is strong evidence to recommend against the use of gefitinib or erlotinib in combination with chemotherapy or as maintenance therapy after chemotherapy and radiation as a first-line treatment for advanced NSCLC. Erlotinib monotherapy is an effective treatment that can prolong survival for patients with advanced NSCLC whose disease has relapsed or recurred after prior chemotherapy. Although a significant survival benefit has not been demonstrated for gefitinib in a placebo-controlled study, the two randomized phase II trials suggest that gefitinib may provide clinically important symptomatic benefits. PMID- 17409887 TI - Management of unresected stage III non-small cell lung cancer: a systematic review. AB - PURPOSE: To conduct a systematic review to determine the most effective therapy for patients with unresected stage III non-small cell lung cancer. METHODS: Relevant randomized trials and meta-analyses were identified through a systematic search of the literature. RESULTS: Forty-seven trials and six meta-analyses were included. No statistically significant survival differences were detected for immediate versus delayed administration of radiotherapy or different doses of hyperfractionated radiotherapy. Three of 12 trials comparing various doses and schedules of radiotherapy detected a statistically significant survival advantage with higher radiation doses. All meta-analyses found a statistically significant survival advantage for chemoradiation, particularly platinum-based, compared with radiation alone. One meta-analysis and three trials comparing concurrent with sequential chemoradiation detected a statistically significant survival advantage with concurrent administration. Increased toxicities, especially esophagitis and hematologic events, were generally associated with concurrent chemoradiation. The survival advantage for concurrent platinum-based chemoradiation corresponds to a 4% absolute survival benefit at 2 years. With respect to trials comparing different chemotherapy regimens or schedules, there is insufficient evidence to determine which particular regimen or schedule is most effective. CONCLUSION: Palliative radiotherapy can provide symptom relief for symptomatic patients with poor performance status. For patients with good performance status, chemoradiation improves survival compared with radiotherapy alone, particularly when the two modalities are administered concurrently. Sequential chemoradiation is a treatment option for borderline-status patients. Adequate assessment of performance status is important when evaluating treatment options for patients with unresected non-small cell lung cancer. Patients and physicians should have a full discussion of the benefits, limitations, and toxicities of therapy. PMID- 17409888 TI - EGFR: a prognostic and/or a predictive marker? PMID- 17409889 TI - Knowledge of efficacy of treatments in lung cancer is not enough, their clinical effectiveness should also be known. AB - The benefits established in efficacy trials, usually randomized, controlled trials conducted under highly controlled circumstances with maximized internal validity, can frequently not be demonstrated in clinical practice at the community level. Effectiveness trials are tools to evaluate the applicability of a treatment in a wider setting with maximized external validity, to observe uncommon adverse events, and to identify factors influencing the main outcomes and risks. Important areas in relation to lung cancer treatment that will benefit from effectiveness trials include gefitinib monotherapy and bevacizumab therapy combined with cytotoxic chemotherapy for advanced non-small cell lung cancer. These therapies were found to produce life-threatening nonhematologic toxicity at a high incidence of up to 5%; however, the risk factors for these toxicities have not yet been fully established. Effectiveness trials of adjuvant chemotherapy after surgery with long-term follow-up are also important to obtain reliable information as to secondary malignancy and noncancer-related deaths. Development of an infrastructure for effectiveness trials is crucial because of the necessity to deal with large numbers of patients, sometimes as many as 10,000 patients, from many hospitals. The extensive research time involved and the considerable cost of these trials may be reduced with the use of Internet resources. Effectiveness trials are a fundamental step toward bridging the gap between clinical research and clinical practice and effectively implementing new therapies in clinical practice. PMID- 17409890 TI - SRC and STAT pathways. PMID- 17409891 TI - EGFR dinucleotide repeat polymorphism as a prognostic indicator in non-small cell lung cancer. AB - BACKGROUND: The epidermal growth factor receptor (EGFR) has been implicated in tumor growth and progression. Intron 1 of the EGFR gene contains a polymorphic simple sequence repeat (SSR) of 14 to 21 CA dinucleotides, the length of which correlates inversely with the level of EGFR transcription. The authors hypothesized that a shorter length of tumor SSR would be associated with poorer survival in patients with non-small cell lung cancer (NSCLC). METHODS: Patients enrolled in Eastern Cooperative Oncology Group E3590 (a randomized, prospective trial of adjuvant therapy following resection of stages II and IIIa NSCLC) were randomized to radiation or radiation plus chemotherapy. Genomic DNA extracted from resected tumors was amplified for EGFR intron 1 by polymerase chain reaction and sequenced in a 3730XL DNA analyzer. RESULTS: One hundred fifty-seven primary tumors were sequenced, 106 (68%) of which were heterozygous for intron 1. The most common genotypes were allele lengths of 17/19 dinucleotides (17.8%), 17/18 (11.4%), and 19/19 (11.4%). Allele status (homozygous versus heterozygous) did not correlate with race, gender, weight, performance status, histology, stage, or survival. Shorter allele length (< or =18 versus >18 CA dinucleotide repeats) was associated with squamous cell histology (p = 0.03). Allele sum of greater than 35 was associated with improved overall survival (log-rank p = 0.03, hazard ratio = 0.66). CONCLUSION: This is the first study to characterize the EGFR intron 1 SSR polymorphism in NSCLC. Tumors were most commonly heterozygous for SSR length. Squamous histology was associated with a shorter SSR. Longer sequences are associated with improved survival. PMID- 17409892 TI - Epidermal growth factor receptor gene mutation and computed tomographic findings in peripheral pulmonary adenocarcinoma. AB - OBJECTIVE: The presence of epidermal growth factor receptor (EGFR) mutations has been reported to predict the response to gefitinib in pulmonary adenocarcinoma patients. A retrospective analysis was conducted to identify the correlation between computed tomographic findings of the nodules and EGFR status. PATIENTS AND METHODS: Computed tomographic findings of 38 patients with peripheral pulmonary adenocarcinoma with EGFR mutations were reviewed and compared with those of 42 peripheral pulmonary adenocarcinoma patients with wild-type EGFR. RESULTS: Mutations were found significantly more frequently among women (28 of 45 women versus 10 of 35 men) and among non-smokers (31 of 47 non-smokers and 7 of 33 smokers). The L858R mutation was found in 18 cases. Several types of deletion mutants in exon 19 were found in 18 cases. The nodules with EGFR mutations (2.5 +/- 1.0 cm) were significantly smaller in diameter than those in the wild-type group (3.1 +/- 1.9 cm). Ground glass opacity (GGO) was more often observed in the mutation group (28 of 38) than in the wild-type group (24 of 42), but the difference was not statistically significant. When mutations were analyzed with reference to both the tumor size and GGO ratio, patients with a tumor < or =3 cm and a GGO ratio > or =50% often had EGFR mutations, and most (10 of 12) were expressed in female patients. No male adenocarcinoma patients with a tumor larger than 4.0 cm had EGFR mutations. CONCLUSIONS: EGFR mutations were found most frequently in small peripheral adenocarcinomas with a GGO ratio > or =50%, especially among women. These factors may be useful in deciding therapeutic strategies for adenocarcinomas when resection or biopsy is not feasible. PMID- 17409893 TI - Gefitinib (IRESSA) with vinorelbine or vinorelbine/cisplatin for chemotherapy naive non-small cell lung cancer patients. AB - This phase I study assessed the safety, pharmacokinetics, and efficacy of gefitinib (IRESSA) combined with vinorelbine or vinorelbine/cisplatin in chemotherapy-naive patients with advanced non-small cell lung cancer (NSCLC). Patients received gefitinib 250 mg/day and vinorelbine (group A; n = 6) or vinorelbine/cisplatin (group B; n = 8). An additional set of group B patients (n = 9) received gefitinib 500 mg/day with vinorelbine/cisplatin. Adverse events were consistent with individual treatments of gefitinib (mild reversible rash, diarrhea) and chemotherapy (asthenia, fever, nausea, vomiting, constipation), although there was a higher than expected incidence of Common Toxicity Criteria grade 3 or 4 hematologic adverse events, specifically febrile neutropenia and neutropenia. Pharmacokinetic data suggested that neither of the chemotherapy regimens affected steady-state exposure to gefitinib and also that steady-state gefitinib did not alter exposure to vinorelbine or cisplatin. Objective, durable antitumor activity was observed: five partial responses (one in group A; four in group B) and six patients with stable disease (all in group B). The safety data demonstrated that gefitinib with vinorelbine or vinorelbine/cisplatin resulted in severe myelosuppression leading to an unacceptable rate of febrile neutropenia. This study does not support the concurrent administration of gefitinib and vinorelbine, with or without cisplatin, as a valid treatment for advanced NSCLC. PMID- 17409894 TI - Short-term restaging of patients with non-small cell lung cancer receiving chemotherapy. AB - OBJECTIVE: To determine whether computed tomography (CT) performed within a month of receiving chemotherapy is useful in assessing response among patients with non small cell lung cancer (NSCLC). PATIENTS AND METHODS: Consecutive patients receiving chemotherapy for NSCLC who underwent short-term CT restaging between April 2001 and June 2005 were included in the study. Serial CT scans were performed within 31 days (mean, 24 days; range, 9-31 days) after receiving chemotherapy for all patients. Tumors were measured in consensus by two diagnostic radiologists. Tumor response was assessed using uni-dimensional tumor measurements according to Response Evaluation Criteria in Solid Tumors criteria. RESULTS: There were 57 patients in the study (30 men, 27 women; mean age, 63 years; age range, 37-85 years). Tumor histology included adenocarcinoma (n = 30), squamous cell carcinoma (n = 17), and NSCLC otherwise unspecified (n = 10). Clinical tumor, node, metastasis stage was stage II (n = 2), stage III (n = 11), and stage IV (n = 44). A significant change in tumor size was observed in eight patients (14%), with tumor regression in two (3%) and progression in six (11%). For these patients, CT scans were performed within 31 days (mean, 25 days; range, 17-31 days). Among the six patients with tumor progression, early detection of therapeutic failure resulted in a change or a discontinuation of chemotherapy for five. CONCLUSION: Early restaging CT is useful in evaluating therapeutic response among patients with NSCLC and may allow the institution of more appropriate therapy. PMID- 17409895 TI - The yield of mediastinoscopy with respect to lymph node size, cell type, and the location of the primary tumor. AB - BACKGROUND: The aim of this study was to investigate the yield of cervical mediastinoscopy (CM) for pathologically diagnosed non-small cell lung cancer (NSCLC), with respect to lymph node size on computed tomography (CT), cell type, and the location of the primary tumor. METHODS: We reviewed 142 patients who underwent standard cervical mediastinoscopy from April of 1994 to June of 2003 for pathologically diagnosed NSCLC. The staging yield was determined by the percentage of "positive" CM (metastatic paratracheal or subcarinal nodes). Mediastinoscopy was performed when the lymph node diameter was >1 cm and, since 2001, whenever paratracheal or subcarinal lymph nodes were clearly seen on CT, regardless of size. Group A includes patients with lymph nodes on CT of > or =1 cm. Group B includes patients with lymph nodes of <1 cm. The chi test was used for statistical analysis. RESULTS: We performed CM on 142 patients. There were 127 men (89.4%) and 15 women (10.6%). The mean age of the patients was 64.7 years (range, 38-83). The global yield was 28.2% (CM positive in 40 of 142 cases). The specific yield by groups was as follows, with respect to lymph node size: group A (lymph nodes >1 cm), 37.9%, and group B (lymph nodes <1 cm), 8.5% (p < 0.01). With respect to cell type, 16.3% were squamous tumors, 42.9% were adenocarcinomas (p < 0.01), 45.5% were large-cell carcinomas, and 44.4% were other types. With respect to the location of the primary tumor and T stage, there were no statistically significant differences (p = 0.09). When only patients with squamous tumors with no enlarged lymph nodes were considered, staging yield was 3.3% (one of 30), and 19.6% (20 of 102) of patients with a negative CM had positive mediastinal lymph nodes at the time of the resection. The yield was 9.8% (10 of 102 patients) in the territory accessible by CM at the time of the resection (true false negative), and 9.8% (10 of 102 patients) in the territory not accessible by CM. CONCLUSION: Lymph node size and cell type of primary tumors should be taken into account when selecting patients for staging with standard MC in NSCLC. In patients with squamous-type tumors with lymph nodes <1 cm on CT, CM could be avoided because its low yield. PMID- 17409896 TI - Phase Ia/Ib chemo-radiation trial of gemcitabine and dose-escalated thoracic radiation in patients with stage III A/B non-small cell lung cancer. AB - INTRODUCTION: The safety of dose-escalated thoracic radiation concurrent with gemcitabine in patients with inoperable stage III non-small cell lung cancer has not been studied. PATIENTS AND MATERIALS: The maximal tolerated dose of 35 mg/m twice-weekly gemcitabine and concurrent standard thoracic radiation was established in a previous phase Ia trial. In this study, a second patient cohort (phase Ib) received twice-weekly gemcitabine concurrent with three-dimensional dose-escalated thoracic radiation (60-74 Gy) after two cycles of induction chemotherapy: gemcitabine (1000 mg/m) day 1 and 8 and carboplatin (area under the curve 5.0-5.5) day 1 every 21 days. RESULTS: Twenty-three patients were entered in the phase Ib portion of this trial. Grade III/IV hematologic toxicity was primarily thrombocytopenia (22%) and neutropenia (26%). Grade III/IV esophageal toxicities occurred in 17% of patients, and grade III radiation pneumonitis/dyspnea was observed in 7 of 23 patients. The median and 2-year survival for phase Ib patients were 17.4 months and 32%, respectively. The overall 1- and 2-year survival for all 39 patients (16 phase Ia, 23 phase Ib) was 69% and 32%, respectively. CONCLUSIONS: Combining 74-Gy thoracic radiation and concurrent gemcitabine is feasible, but the use of this regimen should be limited to the confines of a clinical trial. A randomized phase II trial through the Cancer and Leukemia Group B is underway to further evaluate the efficacy of this regimen. PMID- 17409897 TI - Survival differences by sex for patients with advanced non-small cell lung cancer on Eastern Cooperative Oncology Group trial 1594. AB - INTRODUCTION: Previous data suggest that women may live longer with advanced non small cell lung cancer (NSCLC) than men. We evaluated whether sex affected survival in the Eastern Cooperative Oncology Group (ECOG) E1594 trial. E1594 randomized patients with advanced NSCLC to one of four platinum doublets and found that all four regimens had comparable efficacy. PATIENTS AND METHODS: Patients in the E1594 database were divided into male and female cohorts; response and survival were calculated separately for each cohort. Known prognostic factors and differences in toxicity profiles were compared between the two cohorts. RESULTS: All 1157 eligible patients (431 women, 726 men) from E1594 were included in this analysis. There was no statistically significant difference in performance status, weight loss of >10%, stage, or incidence of brain metastases between women and men. Response rates were similar (19% for both; P = 0.15). The median survival time for women, however, was significantly longer at 9.2 months (95% CI, 8.1-10.4 months) versus only 7.3 months for men (95% CI, 6.8 8.0 months) (P = 0.004 log-rank test). Toxicity was generally greater in women than in men. CONCLUSIONS: Women in ECOG 1594 had a 1.9-month statistically significant improvement in median survival compared with men, despite similar response rates and greater toxicity and no difference in other known prognostic factors. These data strongly support the significance of sex as a separate prognostic factor in advanced NSCLC and emphasize the importance of sex as a stratification factor in future phase III NSCLC trials. PMID- 17409898 TI - Docetaxel in combination with either cisplatin or gemcitabine in unresectable non small cell lung carcinoma: a randomized phase II study by the Japan Lung Cancer Cooperative Clinical Study Group. AB - PURPOSE: To evaluate whether cisplatin-free chemotherapy (docetaxel and gemcitabine [DG]) provides a comparable alternative to cisplatin-based chemotherapy (docetaxel and cisplatin [DC]) as first-line treatment for patients with advanced non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients (n = 133) with stage IIIB to IV NSCLC were randomly assigned to receive DG (docetaxel 60 mg/m, day 8 + gemcitabine 800 mg/m, days 1 and 8, every 3 weeks; n = 65) or DC (docetaxel 60 mg/m, day 1 + cisplatin 80 mg/m, day 1, every 3 weeks; n = 68). The primary end point of the study was overall response rate. No prophylactic use of human recombinant granulocyte colony stimulating factor was allowed. RESULTS: The planned patient number was 150. However, an unexpectedly high incidence of grade 3 interstitial lung disease (11.1%) was identified in the DG arm, so the study was closed early. The overall response rates of the DG and DC arms were 27% and 23.5%, respectively, which demonstrated that the DG treatment was not inferior to the DC arm. Gastrointestinal toxicities were less frequent in the DG arm than in DC arm. Interstitial lung disease was exclusively observed in seven of 63 patients in the DG arm (11.1%). Median survival and 1 year survival rate were comparable between the two arms (median survival, DG 13.7 months versus DC 11.4 months; 1-year survival, DG 56.6% versus DC 47.7%). CONCLUSION: The DG regimen has a response rate and survival rate comparable to those of the DC regimen and can therefore be considered from an efficacy point of view to be comparable. However, the DG regimen may have induced pulmonary toxicity in 11% of the patients exposed and therefore should be used cautiously among patients with advanced NSCLC. PMID- 17409899 TI - Feasibility and validity of a one-item fatigue screen in a thoracic oncology clinic. AB - INTRODUCTION: Fatigue is one of the most common symptoms in patients with advanced cancers. Despite its high prevalence, it is often unrecognized and undetected. This study assessed the feasibility and validity of a one-item fatigue scale (OIFS) in an outpatient oncology clinic. METHODS: Over a 3-month trial period, all patients in a thoracic oncology clinic were screened for fatigue with a one-item, 0 to 10 scale. Over a second trial period, an additional sample of 100 clinic patients completed validated measures of fatigue, including the Functional Assessment of Chronic Illness Therapy-Fatigue Scale (FACIT-F) and Fatigue Symptom Inventory (FSI), in addition to the OIFS. RESULTS: During the initial trial period, more than 95% of patients (574 of 600) had a documented OIFS score on their first clinic visit. Data from the second cohort of patients revealed that the OIFS had good test-retest reliability (r = 0.88) and was highly correlated with the FSI severity scale (0.87) and the FACIT-F (-0.75). Receiver operating characteristic analysis showed the OIFS had good discrimination compared with the FACIT-F (area under the curve = 0.87). Sensitivity and specificity of several OIFS cutoff scores were compared, and scores between 3 and 5 were found to be optimal. CONCLUSIONS: The use of a one-item scale to screen for fatigue is feasible in an ambulatory clinic setting. This scale had convergent validity with other measures of fatigue and was able to identify cases of fatigue that met criteria on the FACIT-F. These data support the recommendations in the National Comprehensive Cancer Network guidelines for cancer-related fatigue screening. PMID- 17409900 TI - Intrapleural urokinase for the treatment of loculated malignant pleural effusions and trapped lungs in medically inoperable cancer patients. AB - BACKGROUND: Pleural fluid loculations or trapped lungs frequently render patients with symptomatic malignant pleural effusions (MPEs) unsuitable for pleurodesis. Thoracoscopic surgery or thoracotomy with decortication is generally not feasible for patients with a poor performance status. MPEs have augmented procoagulant and depressed fibrinolytic activity that contributes to fibrin deposition within the pleural space. The authors conducted an observational prospective cohort study to investigate the use of intrapleural urokinase (IPUK) for such patients and made a comparison with a historical control group. METHODS: Between March of 2000 and August of 2005, 48 consecutive patients with symptomatic MPEs with an average Karnofsky performance scale score of 46.7% were recruited. Dyspnea persisted with the presence of substantial residual loculated MPEs in 36 patients and trapped lungs in 12 patients, when the effectiveness of 8-French intrapleural catheter drainage had decreased despite regular saline flushes. Urokinase was instilled daily through the catheter at a dose of 100,000 IU diluted in 100 ml of normal saline for 3 days. Additional IPUK instillation was required upon partial improvement. The records and chest radiographs of another 52 patients with symptomatic MPEs had met these eligibility criteria between January of 1995 and February of 2000 and received saline flushes only were also reviewed. RESULTS: Immediate lung reexpansion and resolution of dyspnea was achieved in 29 of the 48 patients who underwent IPUK therapy (60.4%). The mean dose of urokinase instillations per patient was 360,000 IU. There were no major complications. A significant association of earlier intervention with the success of IPUK therapy was noted. Responders also had a significantly increased drainage within the 24 hours after the first dose of IPUK. Minocycline pleurodesis was subsequently performed for the 29 IPUK responders. Eighteen patients were followed up until death, with a median survival of 6.5 months. The other remained alive at the time of analysis with a median follow-up of 5 months. Two patients had an immediate failure of pleurodesis at 1 month. Three relapses occurred at 3, 4, and 7 months from pleurodesis, respectively. Twenty-three patients (79.3%) had lifelong pleural symphysis, including 21 having loculated MPEs and two having trapped lungs, respectively. Compared with the historical control group, the IPUK study group had significantly greater improvement on chest radiography and a shorter duration of pleural drainage. CONCLUSION: These results suggest that IPUK is a safe and useful nonsurgical adjunct therapy for loculated MPEs or trapped lungs in medically inoperable cancer patients. PMID- 17409901 TI - Decreased immunoreactivity of CD99 is an independent predictor of regional lymph node metastases in pulmonary carcinoid tumors. AB - BACKGROUND: Few data are available on the prevalence and clinicopathological meaning of CD99, the transmembrane product of the pseudoautosomal MIC2 gene, in pulmonary neuroendocrine tumors. METHODS: We evaluated CD99 immunoreactivity in lung tissues, pulmonary neuroendocrine hyperplasias, and 136 consecutive pulmonary neuroendocrine tumors of diverse histological types. RESULTS: By immunohistochemistry, a membranous and/or cytoplasmic immunoreactivity was seen in 60 of 136 (44%) tumors, whereas both normal and hyperplastic neuroendocrine cells of the lung were consistently nonreactive. A steady decrease of the CD99 labeling index was observed from better to poorly differentiated tumors, with a prevalence of the membranous pattern in typical carcinoids (TCs), and of the cytoplasmic pattern in atypical carcinoids (ACs) and large cell neuroendocrine carcinoma/small cell lung carcinoma (P < 0.0001), independent of tumor stage. In TCs/ACs, increased levels of CD99 labeling index or the membranous pattern were associated with low proliferative fraction (P = 0.0011) and smaller tumor size (P = 0.0054) and with lack of regional lymph node metastases (P = 0.0078). Moreover, CD99 expression decreased according to the pN0-2 classes (P = 0.0016), with an inverse relationship between the number of positive lymph nodes, the labeling index (P = 0.013) and the nonmembranous pattern (P = 0.016). At multivariate analysis, both the decreased CD99 labeling index and the negative/cytoplasmic staining were independent risk indicators for lymph node metastases in the subset of TC/AC patients. No relevant relationships were found in large cell neuroendocrine carcinoma/small cell lung carcinoma. CONCLUSION: CD99 is especially present in low- to intermediate-grade neuroendocrine tumors of the lung, and loss of the marker correlates with the occurrence of nodal metastases in TC/AC patients. PMID- 17409902 TI - Preoperative chemo-radiation-induced ulceration in patients with esophageal cancer: a confounding factor in tumor response assessment in integrated computed tomographic-positron emission tomographic imaging. AB - HYPOTHESIS: Positron emission tomography can be useful in predicting response of esophageal cancer after preoperative chemo-radiation therapy (CRT). We evaluated the use of integrated computed tomography (CT)-PET among patients with esophageal cancer being considered for resection after CRT. METHODS: Three reviewers blinded to clinical and pathologic staging retrospectively reviewed the CT-PET scans of patients with esophageal cancer after preoperative CRT who underwent esophagectomy. [F]-fluoro-2-deoxy-D-glucose uptake for residual malignancy was determined by visual analysis and semi-quantitatively when standardized uptake value (SUV) was > or =4. RESULTS: Forty-two patients underwent esophageal resection. Using visual analysis, CT-PET had a sensitivity of 47% and specificity of 58% in detecting residual malignancy. Using semi-quantitative analysis, 19 patients had a SUV > or =4 in the region of the primary esophageal tumor and were interpreted as having residual malignancy (sensitivity 43%, specificity 50%). Of these 19, six had complete pathologic response to CRT. These false-positive results, due to therapy-induced ulceration detected at endoscopy, limit the use of CT-PET alone in detecting residual malignancy. Similarly, sensitivity (25%) and specificity (73%) of endoscopy/biopsy in detecting residual malignancy were poor. However, the accuracy of CT-PET in detecting residual malignancy was improved when combined with endoscopic findings. In the absence of ulceration at endoscopy, 8 of 8 patients with SUV > or =4 after chemo-radiation had residual malignancy at surgery. CONCLUSIONS: CRT-induced ulceration results in false positive results on CT-PET and precludes accurate detection of residual esophageal tumor. However, CT-PET in combination with endoscopy is useful in identifying patients with a high risk of residual tumor post-CRT. PMID- 17409903 TI - Magnetic resonance imaging changes in synchronous bilateral progressive facial nerve weakness. PMID- 17409904 TI - Photodynamic therapy (PDT) for lung cancers. AB - Photodynamic therapy (PDT), a treatment for cancer, uses a photosensitizer and laser irradiation to produce reactive oxygen in cells. In Japan, the United States, and many other countries, PDT is a treatment option for stage 0 (TisN0M0) and stage I (T1N0M0) centrally located early stage lung cancer. PDT can preserve lung function, can be repeated, and can be combined with other therapeutic modalities such as chemotherapy. Recently, mono-l-aspartyl chlorine e6 (NPe6, Laserphyrin), a second-generation photosensitizer with lower photosensitivity than Photofrin (porfimer sodium), was approved by the Japanese government and a phase II clinical study using NPe6 with a new diode laser demonstrated an excellent antitumor effect and low skin photosensitivity. We expect PDT to be widely employed in many fields and the applications of PDT to be extended because of the decreasing cost of laser equipment and lower systemic photosensitivity induced by the photosensitizer. The purpose of this review is to introduce not only recent clinical trials of PDT for centrally located early lung cancer, but also new applications of PDT for cases of peripheral-type, early-stage lung cancers. We also discuss the applications of PDT for advanced lung cancer and combined therapy using PDT and other treatments for lung cancer. PMID- 17409905 TI - Surgically resected isolated hepatic metastasis from non-small cell lung cancer: a case report. AB - We treated a patient with non-small cell lung cancer (NSCLC) and an isolated hepatic metastasis. He was a 56-year-old male who underwent right pneumonectomy after concurrent chemoradiation therapy (etoposide+cisplatin) with the diagnosis of stage IIIA squamous cell lung carcinoma. Seven months later, an isolated hepatic metastasis was found on a PET-CT scan. Hepatic segmentectomy was performed, and the pathology showed squamous cell carcinoma. Adjuvant chemotherapy with five cycles of gemcitabine and cisplatin was also given. The patient has been followed with PET-CT and CT scanning every 6 months, and there is no evidence of relapse at more than 5 years after the diagnosis of NSCLC. This shows that the surgical resection of an isolated hepatic metastasis may be an option in carefully selected patients with NSCLC without evidence of disease outside the liver. PMID- 17409906 TI - The Liverpool Statement 2005: priorities for the European Union/United States spiral computed tomography collaborative group. AB - The Liverpool Statement 2005 was developed at the Fourth International Lung Cancer Molecular Biomarkers Workshop in Liverpool (October 27-29, 2005) and focused on the priorities for the European Union/United States (EU-US) Spiral Computed Tomography (CT) Collaborative Group. The application of spiral CT technology for early lung cancer screening has gained enormous momentum in the past 5 years. The EU-US Spiral CT Collaboration was initiated in 2001 in Liverpool, and subsequent meetings throughout Europe have resulted in the development of collaborative protocols and minimal data sets that provide a mechanism for the different trial groups to work together, with the ultimate aim to pool results. Considerable progress has been made with major national screening trials in the U.S. and Europe, which include IELCAP, NLST, and NELSON. The major objective of this international collaboration is the planned cross analysis of the individual studies after they are reported. The EU-US researchers have agreed to a number of long-term objectives and to explore strategic areas for harmonization of complementary investigations. PMID- 17409907 TI - The proportion of patients with metastatic non-small cell lung cancer potentially eligible for treatment with bevacizumab: a single institutional survey. PMID- 17409908 TI - Prostacyclin in lung cancer. PMID- 17409909 TI - Pemetrexed alone or in combination with cisplatin in previously treated malignant pleural mesothelioma: outcomes from a phase IIIB expanded access program. AB - BACKGROUND: In a randomized phase III trial, pemetrexed plus cisplatin was associated with improved survival compared with cisplatin alone for patients with malignant pleural mesothelioma (MPM). However, there are limited data available on the efficacy of these and other chemotherapy regimens in patients who have received previous systemic chemotherapy. To gather additional efficacy and safety data of pemetrexed/cisplatin and pemetrexed alone in previously treated patients, we examined patients treated on the Eli Lilly and Company expanded access program (EAP). PATIENTS AND METHODS: Patients with malignant mesothelioma were enrolled in this trial. Of 1056 patients receiving at least one dose of the study drug, 187 (17.7%) were previously treated patients with MPM. Patients were treated every 21 days with pemetrexed 500 mg/m alone (n = 91) or in combination with cisplatin 75 mg/m (n = 96) for a maximum of six cycles. All patients received folic acid and vitamin B12 supplementation and steroid prophylaxis. Serious adverse events (SAEs) were reported by investigators and compiled in a pharmaco vigilance database for all patients enrolled in the EAP. RESULTS: Median age of the previously treated pleural mesothelioma subset was 66 years (range, 27-87 years). Based on 153 evaluable patients (a subset of the larger intent-to-treat population of 187), the overall response rate was 32.5% for pemetrexed and cisplatin and 5.5% for pemetrexed alone. The disease control rate (response rate + stable disease) was 68.7% for pemetrexed and cisplatin and 46.6% for pemetrexed alone. Median survival was 7.6 months for pemetrexed plus cisplatin (67% censored) and 4.1 months for pemetrexed alone (55% censored). The most commonly reported serious adverse events in the overall EAP irrespective of causality were dehydration (7.2%), nausea (5.2%), vomiting (4.9%), dyspnea (3.8%), and pulmonary embolism (2.4%). CONCLUSIONS: The data from this EAP study suggest that patients with previously treated MPM can benefit from treatment with pemetrexed alone or in combination with cisplatin. The treatment is associated with acceptable toxicity. PMID- 17409910 TI - Profiling tumor-associated antibodies for early detection of non-small cell lung cancer. AB - BACKGROUND: A blood test for non-small cell lung cancer (NSCLC) may be a valuable tool for use in a comprehensive lung cancer screening strategy. Here we report the potential of autoantibody profiling to detect early-stage and occult NSCLC. METHODS: T7-phage NSCLC cDNA libraries were screened with patient plasma to identify phage-expressed proteins recognized by tumor-associated antibodies. Two hundred twelve immunogenic phage-expressed proteins, identified from 4000 clones, were statistically ranked for their individual reactivity with 23 stage I cancer patient and 23 risk-matched control samples. All 46 samples were used as a training set to define a combination of markers that were best able to distinguish patient from control samples; this set of classifiers was then examined using leave-one-out cross-validation. Markers were then used to predict probability of disease in 102 samples from the Mayo Clinic CT Screening Trial (six prevalence cancer samples, 40 drawn 1 to 5 years before diagnosis, and 56 risk-matched controls). RESULTS: Measurements of the five most predictive antibody markers in 46 cases and controls were combined in a logistic regression model that yielded area under the receiver operating characteristics curve of 0.99; leave-one-out validation achieved 91.3% sensitivity and 91.3% specificity. In testing this marker set with samples from the Mayo Clinic Lung Screening Trial, we correctly predicted six of six prevalence cancers, 32 of 40 cancers from samples drawn 1 to 5 years before radiographic detection on incidence screening, and 49 of 56 risk-matched controls. CONCLUSIONS: Antibody profiling may be a useful tool for early detection of NSCLC. PMID- 17409911 TI - Predictive factors of gefitinib antitumor activity in East Asian advanced non small cell lung cancer patients. AB - BACKGROUND: Gender, smoking history, adenocarcinoma histology, performance status, and East Asian ethnicity were predictive factors of gefitinib response in previous analysis. However, these factors tend to be correlated with each other; it is not clear whether gender, smoking history, and adenocarcinoma histology were all independent predictors for response in East Asian populations. METHODS: Tumor response, survival and predictive factors of gefitinib response of advanced non-small cell lung cancer patients treated between May of 2002 and November of 2004 were collected retrospectively from three medical centers in Taiwan. Univariate and multivariate logistic regression models were used to test potential predictive factors associated with response to gefitinib. Overall survivals between groups with different predictive factors were compared by log rank tests. Multivariate analyses were performed to identify factors that independently predict for survival. RESULTS: A total of 428 patients were analyzed. The median follow-up duration for living patients was 19.5 months (range, 10.2-39.9). Objective tumor response was observed in 114 patients (26.6%, 95% confidence interval [CI]: 22.4%-30.8%) and disease stabilization in 129 patients (30.2%). Response rate was statistically significant higher in adenocarcinoma, good performance status, and chemonaive patients in multivariate analysis. The median survival was 7.4 months (95% CI: 5.8-9.0) and 1-year survival was 34.3% (95% CI: 29.0%-38.0%). Significant independent predictive factors associated with longer survival in multivariate analysis were good performance status (p < 0.001) and responsiveness to gefitinib (p < 0.001). In 286 chemotherapy-treated patients, the response rate was 22.7%. Median and 1-year survival was 7.9 months and 36.7%, respectively. Good performance status was predictive of tumor response (p < 0.001) and better survival (p < 0.001) in multivariate analysis. Response to gefitinib was predictive of better survival (p < 0.001). CONCLUSIONS: Gender and smoking status were not, but good performance status (PS), no previous chemotherapy, and adenocarcinoma histology were independent predictive factors in multivariate analysis for gefitinib response in Taiwanese advanced non-small cell lung cancer population. In patients previously treated with chemotherapy, only good PS was an independent predictor for tumor response in multivariate analysis. PMID- 17409912 TI - Surgical patterns of care in operable lung carcinoma treated with radiation. AB - PURPOSE: To determine the national surgical practice patterns of care for operable lung cancer patients treated with radiation. MATERIALS AND METHODS: A nationwide survey of a stratified random sample of institutions was conducted for patients who had non-metastatic lung cancer, Karnofsky Performance Scores (KPS) > or =60, and who had received radiation therapy as definitive or adjuvant treatment. Among 541 patients, representing a weighted sample size of 42,335 patients nationwide, 131 (19.8%) underwent surgery as part of their therapy. Pearson chi statistics were used to analyze characteristics of this subset of patients. RESULTS: Of the 131 patients who underwent surgery, 126 patients who had non-small cell lung cancer (NSCLC) were analyzed. Surgical patients were younger, had less weight loss, higher KPS, and higher forced expiratory volume within 1 second (FEV1) values than those treated without surgery. Surgical patients had more stage I/II (53.5% vs 32.2%; p = 0.0004) and less clinical N2/N3 disease (28.8% vs 47.5%; p = 0.002) than nonsurgical patients. Surgery consisted of lobectomy or bilobectomy in 63.2% of patients, pneumonectomy in 23.5%, and wedge resection in 5.9%. Of the patients, 80.4% received radiation in the adjuvant setting and 9.9% in the neoadjuvant setting. CONCLUSIONS: Patients with non-metastatic lung cancer who are treated surgically and with radiation have clinically less advanced disease than those treated with radiation alone. Most radiation therapy in this setting is administrated postoperatively and secondary to hilar and/or mediastinal nodal involvement undetected before surgery. Improved preoperative nodal staging and neoadjuvant approaches may alter these practice patterns. PMID- 17409913 TI - A phase II study of induction therapy with carboplatin and gemcitabine among patients with locally advanced non-small cell lung cancer. AB - INTRODUCTION: The objectives of this trial were to evaluate the activity and safety of gemcitabine carboplatin as induction therapy in patients with locally advanced non-small cell lung cancer METHODS: Patients received two cycles of gemcitabine (1250 mg/m on day 1 and 8), plus carboplatin (area under the curve = 5 on day 1), after which response was established. Patients received a third course only in the case of an objective response (OR). Non-responding patients were directly irradiated. Toxicity was assessed according to the NCI-CTC version 2, radiation toxicity was assessed according to RTOG criteria. Response evaluation was performed according to RECIST criteria. RESULTS: We identified 42 patients, of whom 37 were eligible. Of these, 51% (95% CI, 34%-68%) achieved an OR, all partial responses. No disease progression on therapy was established. Toxicity was mostly hematological: 35% trombocytopenia grade 3 and 4, and 40% neutropenia grade 3 and 4. No severe bleeding or hospitalization because of febrile neutropenia occurred. CONCLUSIONS: Gemcitabine and carboplatin administered according to a 3-week schedule is an active and safe induction regimen. Pending the results of a phase III study, we believe that it is a reasonable alternative among patients for whom cisplatin-based chemotherapy is contraindicated. PMID- 17409914 TI - Sequential vinorelbine and docetaxel in advanced non-small cell lung cancer patients age 70 and older and/or with a performance status of 2: a phase II trial of the Southwest Oncology Group (S0027). AB - BACKGROUND: This phase II study (S0027) evaluated the efficacy and tolerability of planned sequential single-agent chemotherapy with vinorelbine followed by docetaxel in patients with advanced non-small cell lung cancer (NSCLC) age 70 and older and/or a performance status (PS) of 2. METHODS: Patients with stage IIIB (pleural effusion) or stage IV NSCLC, age 70 and older with a PS of 0-1 or 2, any age, received three cycles of vinorelbine 25 mg/m days 1 and 8 every 21 days followed by three cycles of docetaxel 35 mg/m days 1, 8, and 15 every 28 days. RESULTS: A total of 125 patients entered the study; 117 patients were assessable for response, survival, and toxicity. Seventy-five patients were in stratum1 (age 70 and older, PS 0-1) and 42 patients in stratum 2 (PS 2, any age). Objective response was 19% (95% confidence interval [CI]: 11%-30%) and 11% (95% CI: 3%-25%) in strata 1 and 2, respectively. Median survival was 9.1 months (95% CI: 7.1 12.7) and 5.5 months (95% CI: 3.1-6.5) in strata 1 and 2, respectively. Survival at 12 months was 41% and 13% in strata 1 and 2, respectively. Grade 3/4 neutropenia was seen in 32% and 31% of patients in strata 1 and 2, respectively. Three deaths probably related to treatment were noted: one in stratum 1 and two in stratum 2. CONCLUSION: Sequential vinorelbine and docetaxel is a well tolerated and effective regimen in comparison with reports of other treatments tested in patients with advanced NSCLC age 70 and older and/or with a PS of 2. PMID- 17409915 TI - Salvage therapy for Chinese non-small cell lung cancer patients who failed previous chemotherapy. AB - Our aim was to determine the appropriate salvage regimen for Chinese non-small cell lung cancer (NSCLC) patients who failed previous chemotherapy. We retrospectively analyzed data from our seven clinical trials, including single agent gemcitabine, gefitinib, docetaxel with a different schedule, vinorelbine plus cisplatin, vinorelbine plus gemcitabine, docetaxel plus gemcitabine, and docetaxel plus ifosfamide, with a total of 342 cases (including 314 patients, of which 28 entered two different trials due to different salvage line settings), and compared these data with those of other studies, addressing the efficacy and toxicity of salvage therapy in patients who failed previous chemotherapy to analyze choosing of an appropriate salvage regimen. Of the 342 cases receiving salvage treatment, 71.1% were in second-line treatment, and 28.9% in third-line or later treatment. The response rate to our salvage therapy ranged widely, from 6.1% to 36.1%. Median survival was between 5.7 and 8.4 months when different salvage chemotherapy regimens were used, whereas it was 9.3 months in those who received gefitinib treatment. Similarly, 1-year survival ranged between 19.7% and 40% in a chemotherapy setting, and 40.8% for gefitinib treatment. Those who received gefitinib had better toxicity profiles than those who received other regimens. Febrile neutropenia occurred in 19 patients who received a chemotherapy agent (6.2%), and four patients died despite granulocyte colony-stimulating factor and antibiotic treatment. Grade 3 or 4 interstitial pneumonitis occurred in 14 of 247 patients (5.7%) who received docetaxel with/without another agent, and 10 patients died. Grade 3 interstitial pneumonitis occurred in one patient who received gefitinib treatment and recovered. In conclusion, both chemotherapeutic agents, such as docetaxel alone or gemcitabine plus vinorelbine, and gefitinib, are probably appropriate salvage regimens for Chinese NSCLC patients who have failed previous chemotherapy. However, gefitinib has a better safety profile and probably better survival than the chemotherapeutic agents and would be an appropriate alternative choice for salvage chemotherapy, even in a second-line setting for Chinese patients. PMID- 17409916 TI - A phase I study of amrubicin combined with carboplatin for elderly patients with small-cell lung cancer. AB - BACKGROUND: Amrubicin, a new anthracycline agent, has shown high activity for small-cell lung cancer (SCLC) with acceptable toxicities in previous studies. However, a combination regimen of platinum and amrubicin for elderly patients has not been reported. In this phase I study, the dose-limiting toxicity (DLT), the maximal tolerable dose (MTD), and the antitumor activity of a combination of amrubicin and carboplatin in elderly patients with SCLC were evaluated. PATIENTS AND METHODS: Previously untreated elderly patients (> or =70 years old) with SCLC were enrolled in this study. Amrubicin was administered from day 1 to day 3, and carboplatin was administered on day 1 intravenously. The treatment was repeated every 3 weeks. Three escalating dose levels of amrubicin (mg/m)/carboplatin (area under the curve; AUC) (40/4.0, 40/5.0, and 45/5.0) were initially planned. RESULTS: Twelve patients were enrolled. At level 1 (amrubicin 40 mg/m and carboplatin AUC 4.0), all three patients experienced DLTs (grade 4 neutropenia > or =4 days, thrombocytopenia <20,000/mm, or grade 3 diarrhea), and this dose level was determined to be the MTD. At the reduced dose of level 0 (amrubicin 35 mg/m and carboplatin AUC 4.0), although DLTs were observed in three of the nine patients, they were considered to be clinically not severe and could be managed. Non-hematological toxicities were mild or moderate and reversible. The objective response rate was 83%, and the median survival time was 12.7 months. CONCLUSIONS: The MTD of this combination was amrubicin 40 mg/m and carboplatin AUC 4.0, and the recommended dose for a phase II trial is a combination of amrubicin 35 mg/m and carboplatin AUC 4.0. We are now conducting a multicenter phase II trial of this regimen to determine the activity of this combination for elderly patients with SCLC. PMID- 17409917 TI - Impact of pretreatment factors on adverse events: a pooled analysis of North Central Cancer Treatment Group advanced stage non-small cell lung cancer trials. AB - INTRODUCTION: This pooled analysis was performed to examine the impact of pretreatment factors on severe (grade 3 or higher) adverse events (AE) in patients with advanced stage non-small cell lung cancer (NSCLC). METHODS: A pooled data set of 1053 participants from nine North Central Cancer Treatment Group clinical trials was used. Age, gender, performance status, tumor stage, body mass index, serum creatinine levels, hemoglobin levels, white blood cell counts, and platelet counts were evaluated univariately and multivariately using logistic regression. The magnitude of the effects of the pretreatment factors after adjusting for type of chemotherapy agent (platinum versus no platinum) was explored in the final multivariate model. RESULTS: Women and older participants had a significantly greater risk for experiencing severe hematologic and non hematologic AE. Participants with performance status >0 had an increased risk for severe non-hematologic AE. For every one unit (10/L) increase in pretreatment white blood cell count, there was an 11% reduction in the odds of experiencing a severe hematologic AE. The magnitude of these effects on the end points remained similar after adjusting for type of chemotherapy agent. CONCLUSIONS: Pretreatment factors of gender, age, performance status, and hematologic parameters were significant predictors of severe AE among patients with advanced stage NSCLC. This suggests the need to control or adjust for factors that predispose patients to an increased risk of severe AE. These findings can aid in tailoring therapy to individual patients and in the proper design of future clinical trials. PMID- 17409918 TI - Does granulocyte colony-stimulating factor affect survival in patients with advanced non-small cell lung cancer? AB - BACKGROUND: Platinum-based chemotherapy is standard treatment for patients with advanced lung cancer. The common side effect of this therapy is myelosuppression, for which different stimulating factors are used. In this article, the effect of granulocyte colony-stimulating factor (G-CSF) administration on the survival of patients with unresectable non-small-cell lung cancer (NSCLC) was evaluated. METHODS: The charts of 127 patients, treated with carboplatin-based chemotherapy, were reviewed for histology, stage, performance status, weight loss, treatment regimen, toxicity, and survival. Eighty patients were stage IIIA/IIIB NSCLC; 47 were stage IIIB (pleural effusion) or stage IV. Eighty-one patients (63%) experienced severe (grades 3 and 4) neutropenia. Forty-two patients received G CSF, 37 patients for severe neutropenia (14 with febrile neutropenia) and five patients for active infection during chemotherapy. RESULTS: Preliminary analyses, both unadjusted (median survival, 20 months versus 13.8 months; log-rank test, p = 0.02) and adjusted for covariates of interest (Cox regression, hazard ratio = 0.62, p = 0.03) showed a significant effect of the use of G-CSF on survival, even though the groups were balanced with respect to stage, performance status, weight loss, and dose intensity of chemotherapy. Patients with grades 3 and 4 neutropenia (whether they received G-CSF or not) had a better survival outcome compared to those who did not have neutropenia (median survival, 17.6 months versus 11.9 months, log-rank test, p = 0.04). A landmark analysis showed a marginally significant effect of G-CSF on survival (median survival, 18.6 months versus 15.1 months, log-rank test, p = 0.08), even after adjustment for covariates. The Cox regression with the use of G-CSF defined as a binary time varying covariate also showed similar results (Cox regression, hazard ratio = 0.67, 95% CI: 0.42-1.04, p = 0.07). CONCLUSION: In this study, the time bias due to the delayed administration of G-CSF contributed to the longer survival of patients receiving G-CSF. Prospective studies are required to determine whether G CSF has any effect on survival in patients with advanced NSCLC. PMID- 17409919 TI - Impact of skeletal complications on total medical care costs among patients with bone metastases of lung cancer. AB - INTRODUCTION: Previous studies have estimated the costs of skeletal-related events (SREs) for patients with bone metastases of solid tumors by tallying costs for services specifically attributable to these events. This approach may underestimate costs if SREs indirectly increase use of other services. METHODS: This is a retrospective observational study using a large health insurance claims database. Patients with bone metastases of lung cancer who experienced > or =1 SRE were matched to similar patients without SREs based on propensity scores. Kaplan-Meier estimated total medical care costs were compared for propensity matched samples of patients with SREs and without SREs. RESULTS: We identified 534 patients with lung cancer and bone metastases, including 295 (55%) with > or =1 SRE. After matching, there were 162 patients each in the SRE and no-SRE groups with mean follow-up of 5.3 and 3.9 months, respectively. In the SRE group, costs of treatment of SREs were $9,480 (95% CI $7,625 to $11,374) per patient. Total medical care costs were $27,982 (95% CI $15,921 to $40,625) greater for SRE versus no-SRE patients (p < 0.001). CONCLUSIONS: The costs of SREs in patients with lung cancer and bone metastases are substantial and potentially greater than previously estimated. PMID- 17409920 TI - Risk of lung cancer and past use of cannabis in Tunisia. AB - The association between the use of cannabis and the risk of lung cancer is unclear. A hospital-based case-control study was conducted among men in Tunisia and included 149 incident lung cancer cases and 188 controls. Tobacco smoking was significantly associated with an increased risk of lung cancer with odds ratios increasing linearly (p for trend < 0.0001) from 3.9 (95% confidence interval [CI], 1.4-10.9) for former smokers to 17.1 (95% CI: 6.3-46.3) among current smokers who had smoked for >35 years. The odds ratio for the past use of cannabis and lung cancer was 4.1 (95% CI: 1.9-9.0) after adjustment for age, tobacco use, and occupational exposures. No clear dose-response relationship was observed between the risk of lung cancer and the intensity or duration of cannabis use. This study suggests that smoking cannabis may be a risk factor for lung cancer. PMID- 17409921 TI - Multiple pulmonary nodules in an elderly woman. PMID- 17409922 TI - Salvage therapy in patients with advanced non-small cell lung cancer. AB - Patients with advanced non-small cell lung cancer continue to have a poor prognosis; most die from the disease within 1 year. Chemotherapy is beneficial for some patients in the first-line metastatic setting. Three agents, namely docetaxel, pemetrexed, and erlotinib, are approved by the United States Food and Drug Administration as treatment in the second-line setting. In this article, we examine the data supporting the use of these agents in the second-line setting and review data from other completed trials. Lastly, we propose strategies to advance the treatment of patients with non-small cell lung cancer. PMID- 17409923 TI - Solitary fibrous tumor of the pleura with associated hypoglycemia: Doege-Potter syndrome: a case report. AB - Tumor-associated hypoglycemia as a paraneoplastic phenomenon is a well-known entity and is referred to as Doege-Potter syndrome. A man was admitted because of acute confusion and drowsiness. Laboratory results showed profound hypoglycemia. All investigations proved to be normal, except for a chest x-ray, which showed a large pleural mass. On transthoracic puncture, a tumor of pleural origin was diagnosed. This tumor, presenting as a large, well-circumscribed encapsulated mass, was removed by thoracotomy. On pathologic examination, the diagnosis of a solitary fibrous tumor with benign characteristics was made. After surgical removal, the hypoglycemia resolved. Solitary fibrous tumors are localized tumors of the pleura with an unpredictable behavior. The therapy consists of resection. PMID- 17409925 TI - Induction chemotherapy: a surgeon's perspective. PMID- 17409924 TI - The use of chemotherapy in patients with advanced malignant pleural mesothelioma: a systematic review and practice guideline. AB - BACKGROUND: This clinical practice guideline, based on a systematic review, was developed to determine which chemotherapeutic agents (or combinations of agents) show the highest response rates, improved survival, quality of life, or symptom control in patients with advanced malignant pleural mesothelioma. METHODS: A thorough systematic search of the literature was conducted for published articles and conference proceedings for applicable abstracts. Relevant trials, published as articles and abstracts, were selected and assessed. External feedback was obtained from Ontario clinicians, and the guideline was approved by the provincial Lung Cancer Disease Site Group. RESULTS: One hundred nineteen studies were eligible, including eight randomized trials and 111 phase II trials. The pooled response rates from phase II trials suggest that response rates with combination chemotherapy are higher than with single agents. Data from the largest randomized controlled trial demonstrated that chemotherapy with cisplatin and pemetrexed significantly improves response rates (41% versus 17%, p < 0.001), time to progression (5.7 months versus 3.9 months, p = 0.001), and overall survival (median, 12.1 months versus 9.3 months, hazard ratio = 0.77, p = 0.020) in comparison to single-agent cisplatin. A second trial demonstrated cisplatin and raltitrexed significantly improved median survival compared to single-agent cisplatin (11.4 months versus 8.8 months; hazard ratio = 0.76, p = 0.0483). Overall response rate (24% versus 14%, p = 0.056) was greater in the combination treatment arm, but this difference was not statistically significant. CONCLUSIONS: There is good evidence to recommend chemotherapy with pemetrexed and cisplatin for adult patients with symptomatic advanced malignant pleural mesothelioma. Such treatment should be administered with supplementation of vitamin B12 and folic acid. If pemetrexed is not available, cisplatin plus raltitrexed is a reasonable alternative. PMID- 17409926 TI - Insulin-like growth factor and lung cancer. AB - Lung cancer is the leading cause of cancer-related death in the United States. Despite the availability of several cytotoxic and a few molecularly targeted agents, the outlook for patients with advanced non-small cell lung cancer continues to be dismal. Novel approaches are desperately needed. The insulin-like growth factor (IGF) pathway plays an important role in a number of human malignancies contributing to unregulated cell proliferation. The IGF pathway has several targets for therapeutic intervention. Preclinical studies of IGF inhibitors have demonstrated synergism when combined with chemotherapy agents and radiation. Clinical studies are currently ongoing to investigate the safety and efficacy of IGF inhibitors in combination with chemotherapy agents. In this review, we discuss the biology of the IGF pathway and various potential targets for therapy. PMID- 17409927 TI - A systematic review and meta-analysis of the literature: chemotherapy and surgery versus surgery alone in non-small cell lung cancer. AB - BACKGROUND: The effectiveness of preoperative chemotherapy in the treatment of non-small cell lung cancer has remained unclear despite the conduct of several randomized controlled trials (RCTs). METHODS: A systematic review and meta analysis was carried out to assess the effectiveness of preoperative chemotherapy in non-small cell lung cancer. This involved identifying eligible RCTs and extracting aggregate data from the abstracts or reports of these RCTs. Hazard ratios were calculated from these published summary statistics and then combined to give pooled estimates of treatment efficacy. RESULTS: Twelve eligible RCTs were identified, from which data from seven RCTs, including 988 patients (75% of eligible patients), could be combined in a systematic review and meta-analysis. Preoperative chemotherapy improved survival with a hazard ratio of 0.82 (95% confidence interval, 0.69-0.97; p = 0.02). This is equivalent to an absolute benefit of 6%, increasing overall survival across all stages of disease from 14% to 20% at 5 years. There was no evidence of statistical heterogeneity. CONCLUSIONS: This analysis shows a significant benefit of preoperative chemotherapy and is currently the best estimate of the effectiveness of this therapy, but this is based on a small number of trials and patients. This current analysis was unable to address important questions such as whether particular types of patients may benefit more or less from preoperative chemotherapy or whether the early stopping of a number of included RCTs impacted on the results. To assess this, an individual patient data meta-analysis is required. PMID- 17409928 TI - The signature from messenger RNA expression profiling can predict lymph node metastasis with high accuracy for non-small cell lung cancer. AB - BACKGROUND: The extent of regional lymph node (LN) metastasis is the most important factor in the evaluation of resectability and prognosis of non-small cell lung cancer (NSCLC) to increase the chance of complete cure. The authors attempted to deduce a group of genes from the analysis of mRNA expression profiles of the tumor tissues of NSCLC patients with or without LN metastasis, and make a classification model for better prediction of LN metastasis. METHODS: The authors analyzed mRNA expression profiles of 79 NSCLC patients with or without LN metastasis, and deduced the gene signature for the predictive model of LN metastasis in lung cancer. The authors evaluated the predictive accuracy of each of four algorithms by applying them to another set of 33 NSCLC patients. Each algorithm's accuracy was calculated by 10-fold cross-validation, and a combined model showed a level of accuracy that was higher than any one of the better three component algorithms (i.e., ANN, DT, or NB). Avadis, SAS, ArrayXPath, and R-package were the statistical analysis software packages used. RESULTS: The authors selected 949 genes using a classical permutation t test (p < 0.01) and finally obtained a gene signature consisting of 31 genes by adjustment of multiple-hypothesis testing. The LN metastasis prediction model derived from the signature (31 genes) and their characteristic interactions provided a predictive accuracy of 84.85% when applied to a test set of 33 patients. CONCLUSIONS: The authors have demonstrated that their gene signature developed by the expression profiling of mRNAs from the primary tissue could predict the LN metastasis status of NSCLC. PMID- 17409929 TI - PTEN and PIK3CA expression is associated with prolonged survival after gefitinib treatment in EGFR-mutated lung cancer patients. AB - BACKGROUND: We and other researchers have previously reported that pulmonary adenocarcinomas with epidermal growth factor receptor (EGFR) mutations are usually sensitive to gefitinib, an EGFR-specific tyrosine kinase inhibitor, although this relationship is not complete. In this study, we searched for mutations or changes in the expression of genes downstream to EGFR and evaluated their relationship with the effectiveness of gefitinib. METHODS: We studied 78 lung cancer patients who had recurrent disease after surgical resection and were treated with gefitinib. We searched for mutations occurring in the KRAS and PIK3CA gene. We also evaluated the expression level of EGFR, PIK3CA, and PTEN by real-time reverse transcriptase polymerase chain reaction. Gefitinib effectiveness was evaluated by imaging studies; a survival analysis was also done. RESULTS: We found seven (9%) somatic mutations in KRAS and two (2%) in PIK3CA. EGFR mutations were present in 44 (56%). KRAS mutations were found only in tumors without EGFR mutations, whereas PIK3CA mutation was found in tumors with EGFR mutation. Tumor response was assessable in 52 tumors. None of the six tumors with KRAS mutations responded to gefitinib treatment; however, two tumors with PIK3CA mutations showed partial response. Survival was significantly longer in patients with EGFR mutations or in those without KRAS mutations. In tumors with EGFR mutations, survival was longer in those with high PIK3CA or PTEN expression than in those with low expression of these molecules. CONCLUSIONS: An evaluation of the KRAS mutation, as well as PIK3CA and PTEN expression, might help identify lung cancer patients who are most suitable for gefitinib treatment. PMID- 17409930 TI - Mutation and polymorphism in the EGFR-TK domain associated with lung cancer. AB - BACKGROUND: The epidermal growth factor receptor (EGFR) is involved in the development and progression of lung cancer. Somatic EGFR mutations are predictors of response to treatment with EGFR tyrosine kinase (TK) inhibitors (TKIs) for lung cancer, especially among never smokers. EGFR mutations may occur independently of other genetic alterations. METHODS: The authors sequenced the EGFR-TK domain and the K-ras and p53 genes from lung tumor tissues from 44 never smokers and 46 smokers. A case-control study also was conducted to examine the relationship between an EGFR single nucleotide polymorphism in the TK domain and the lung cancer through a multivariate logistic regression analysis. In addition, the authors compared cell growth kinetics, EGFR-TKI sensitivity by MTT, and activation of signaling molecules by immunoblot in lung cancer cell lines with and without EGFR-TK mutations. RESULTS: EGFR-TK mutations were more frequently observed in never smokers (25%) than in smokers (2.2%) (p = 0.001). Excluding cases with a K-ras mutation, the frequency of EGFR-TK domain mutation was still significantly higher in never smokers than in smokers, 26.2% versus 4.5% (p = 0.046). EGFR-TK mutations and K-ras mutations (p = 0.015), and p53 and K-ras mutations (p = 0.015) were mutually exclusive, but p53 and EGFR-TK mutations were not (p = 1.00). During sequencing of the EGFR-TK domain in tumors, an EGFR polymorphism (G2607A) was identified. The genotype AA and AA + AG occurred at a significantly higher frequency in lung cancer cases (n = 122) when compared with controls (n = 147) (odds ratio, 3.39 and 2.67; 95% confidence interval, 1.41-8.17 and 1.17-6.08, p = 0.006 and p = 0.02, respectively). This polymorphism was found independently of EGFR-TK mutations in lung cancer cases, indicating that it does not predispose to mutations. In vitro, lung cancer cell lines with EGFR-TK mutations also did not contain K-ras mutations and displayed a lower growth rate (50%, p = 0.013) than EGFR-TK wild-type cell lines. EGFR-TK mutant cell lines were more sensitive to both gefitinib and erlotinib, although relative sensitivity to erlotinib compared with wild-type was less pronounced than for gefitinib. Cell lines with a lower growth rate also expressed higher levels of E cadherin than faster growing cell lines. CONCLUSIONS: EGFR-TK mutation frequency is high in never-smoking lung cancer patients and is exclusive of mutation in K ras but not p53. In addition to somatic EGFR-TK mutations that arise in lung tumors, germline variation in the EGFR-TK domain might also be associated with an increased risk of lung cancer. Somatic EGFR-TK mutations alter cell biology and response to EGFR-TKIs and may be mutation specific. PMID- 17409931 TI - Manganese superoxide dismutase and catalase genetic polymorphisms, activity levels, and lung cancer risk in Chinese in Hong Kong. AB - INTRODUCTION: Antioxidants play an important role in counteracting the effects of potential carcinogens. We investigated the risk of lung cancer development with respect to manganese superoxide dismutase (MnSOD) and catalase genetic polymorphisms and their association with erythrocyte antioxidant activities. PATIENTS AND METHODS: This was a case-control study involving patients with confirmed lung cancer and age-matched healthy controls. Genotyping of MnSOD and catalase in DNA extracted from peripheral white cells was performed by polymerase chain reaction-based restriction fragment length polymorphism. Erythrocyte superoxide dismutase and catalase activities were measured spectrophotometrically using chemical kinetic reactions. RESULTS: We recruited 240 patients with lung cancer (63% male, aged 55.6 +/- 11.9 years, 58% adenocarcinoma, 85% clinical stage III or IV) and 240 age-matched healthy controls. The frequencies of the Val allele of MnSOD gene and the C allele of catalase gene were common (>86% and 90%, respectively), with similar distribution, in both patients with lung cancer and controls. The homozygous variant genotypes of MnSOD and catalase were not associated with increased lung cancer risk. The erythrocyte SOD and catalase activity was significantly lower among all patients with lung cancer as a whole compared with controls, irrespective of genotypes. However, patients with adenocarcinoma and non-adenocarcinoma showed differences in SOD and catalase activity among different genotypes in comparison with controls. CONCLUSION: The common Val16Ala MnSOD polymorphism and C-T substitution in the promoter region of the catalase gene do not confer increased or reduced risk of lung cancer in Chinese in Hong Kong. PMID- 17409932 TI - An ecological study of the association of metal air pollutants with lung cancer incidence in Texas. AB - BACKGROUND: Air pollution particulate matter and tobacco smoke, which contain metals that are human lung carcinogens, are associated with lung cancer risk. We conducted an ecological study to examine the association of metal air pollutants with lung cancer incidence in Texas. METHODS: During the period 1995 to 2000, 81,132 lung cancer cases were reported in Texas. We identified eight metals that (1) are in airborne particulate matter or tobacco smoke or are human lung carcinogens and (2) had consistent Environmental Protection Agency air release reporting for multiple counties from 1988 through 2000. We examined the association of metal air releases with the average annual age-adjusted primary and non-small cell lung cancer rates in the 254 Texas counties. RESULTS: Univariate analysis indicated the following positive associations: (1) zinc with the primary (p = 0.02) and non-small cell (p < 0.01) lung cancer rates and (2) chromium and copper with the non-small cell lung cancer rate, p = 0.01 and p = 0.01, respectively. In the multivariate analyses, risk adjusted for sex, race and ethnicity, and urbanization, zinc was positively associated with the primary (beta = 0.13, p = 0.01) and non-small cell (beta = 0.14, p = 0.02) lung cancer rates, and when interaction terms among the eight metals were included, zinc was significantly and positively associated with these rates. Smoking prevalence was similar for counties with and without releases for the eight metals. CONCLUSIONS: The study suggests that inhalation exposure to metals, including those that are essential human nutrients, play a role in lung carcinogenesis. PMID- 17409933 TI - Gemcitabine, cisplatin, and hyperfractionated accelerated radiotherapy for locally advanced non-small cell lung cancer. AB - BACKGROUND: Due to potent radiosensitization and potential serious or fatal toxicity, concurrent gemcitabine and irradiation should only be applied within clinical trials. We here present experience from a phase I-II clinical trial for patients with locally advanced non-small cell lung cancer (NSCLC) treated with hyperfractionated accelerated radiotherapy and concurrent low-dose gemcitabine. METHODS: Eligible patients had locally advanced inoperable NSCLC without pleural effusion, Eastern Cooperative Oncology Group performance status 0-1, were chemotherapy naive and had no previous radiotherapy to the chest, and had adequate hematopoietic, liver, and kidney function. Routine brain computed tomography was not performed, and positron emission tomography/computed tomography was not available. Treatment consisted of three parts: induction chemotherapy with gemcitabine and cisplatin in standard doses, local treatment with concurrent chemotherapy and radiotherapy, and consolidation chemotherapy. Patients were irradiated with opposed AP-PA and oblique fields, using 2.5-D treatment planning. Although corrections for inhomogeneous tissue were made, volume of total lung receiving > or =20 Gy (V20) could not be determined. The trial started as phase I, aimed to determine the dose-limiting toxicity and maximal tolerated dose (MTD) for concurrent hyperfractionated radiotherapy (1.4 Gy twice daily) and gemcitabine 55 mg/m twice weekly as a radiosensitizer. Phase II of the trial then continued at the level of MTD. RESULTS: Twenty-eight patients with NSCLC, nine patients with stage IIIA, 16 patients with IIIB, and three patients with an inoperable recurrence after previous surgery, entered the trial. The first 12 patients entered Phase I of the trial at the initial level of 42 Gy in 30 fractions in 3 weeks. Dose-limiting toxicity was acute esophagitis; 47.6 Gy in 34 fractions in 3.5 weeks was the MTD for this regimen of concurrent chemotherapy and radiotherapy. In phase II of the trial, this dose was applied to the next 16 patients. Among all 28 patients, 13 had grade 3 or 4 acute toxicity: esophagitis (eight patients), neutropenia (eight patients), thrombocytopenia (four patients), and anemia (two patients). No pulmonary toxicity and no persistent or serious late toxicity were seen. Local and/or regional relapse was documented in nine patients, distant in five and both locoregional and distant in 10 patients. The most common sites of distant spread were the brain and lung in eight and six patients, respectively. At 2 years, progression-free survival was 43% and overall survival was 57%. After 43 to 85 months of follow-up, seven patients are alive, of whom six (21%) are without evidence of disease and may be regarded as long-term survivors. Among the long-term survivors, one was in the group irradiated to 42 Gy and six in the groups irradiated to 47.6 Gy. CONCLUSION: Judging from current standards, the methods used in diagnostics and in planning of radiotherapy were suboptimal. Using modern radiotherapy planning, a higher MTD, possibly a different profile of toxicity, and better long-term results may be expected. The high incidence of brain relapse emphasizes the need for careful screening for unsuspected brain disease before treatment and the importance of clinical studies on prophylactic cranial irradiation for patients with locally advanced NSCLC. Although the small number of patients in this study precludes any definitive conclusion, it appears that our program of concurrent chemotherapy and radiotherapy offers a chance for disease control at least comparable to previously described programs for inoperable lung cancer. PMID- 17409934 TI - Stage is not a reliable indicator of tumor volume in non-small cell lung cancer: a preliminary analysis of the Trans-Tasman Radiation Oncology Group 99-05 database. AB - BACKGROUND: Tumor volume has been shown to be a prognostic factor for the response of some tumors to radiotherapy. TNM stage has prognostic value for patients treated surgically for non-small cell lung cancer (NSCLC), but its value is less clear for patients treated by nonsurgical means. This may be because tumor size is not a consistent determinant of T stage or stage group. As part of the preliminary analyses for the Trans-Tasman Radiation Oncology Group 99-05 study, the authors performed this analysis to determine to what extent stage reflects tumor volume. METHODS: In this prospective multicenter observational study, patients had to have histologically proven NSCLC, no evidence of disease beyond the primary site or thoracic lymph nodes, and been planned for radical radiotherapy with or without chemotherapy. Tumor volume measurements were based on computed tomography-based treatment planning images. RESULTS: Four hundred four patients were available for analysis. There was a strong correlation between (log) maximum tumor diameter and (log) tumor volume (r = 0.93, p < 0.001). Although there was a highly significant trend of increasing volume with increasing T stage and stage group, when tumors were categorized into four groups according to increasing volume, there was only 55% concordance with T stage and 67% concordance with stage group. CONCLUSIONS: There is limited correlation between tumor size and disease stage in patients with NSCLC. This justifies documentation and investigation of size as a potential prognostic factor independent of stage. Maximum tumor diameter may be an adequate substitute for volume as a measurement of size. PMID- 17409935 TI - A phase II trial of carboplatin and gemcitabine with exisulind (IND #65,056) in patients with advanced non-small cell lung cancer: an Eastern Cooperative Oncology Group study (E1501). AB - BACKGROUND: Carboplatin and gemcitabine are one standard regimen for patients with advanced non-small cell lung cancer (NSCLC). The oral proapoptotic agent exisulind is a cyclic guanosine monophosphate phosphodiesterase that increases apoptosis in vitro. We performed a phase II trial of carboplatin and gemcitabine with exisulind in patients with advanced NSCLC. METHODS: Gemcitabine (1000 mg/m days 1 and 8) and carboplatin (AUC = 5 day 1) were administered every 21 days, with exisulind orally at 250 mg orally twice daily continuously, starting day 1. The primary objective was to evaluate the 18-month survival. Secondary objectives included response rate, progression-free survival, and toxicities. Eligibility included stage IIIB (pleural effusion) or stage IV NSCLC, no previous chemotherapy, and an Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0-1. RESULTS: Of 57 eligible patients treated, 34 patients were male and 23 female, 42 had stage IV, six stage IIIB, and nine had recurrent disease. The median age was 63 years (range, 37-83). Twenty-six patients had an ECOG PS of 0 and 31 had a PS of 1. The majority of grade 3-4 toxicities were hematologic. Grade 3-4 nonhematologic toxicity seen in >5% of patients included nausea/vomiting in 16% and fatigue in 23% of patients. The overall response rate was 19.3%. Median progression-free survival was 4.7 months. Median overall survival was 9.0 months. Eighteen-month overall survival was 30%. CONCLUSION: The chemotherapy combination of gemcitabine and carboplatin with the oral proapoptotic agent exisulind is generally well tolerated with principally hematologic toxicity. The statistical endpoint of 17 patients alive at 18 months was met, but given ongoing developments in advanced NSCLC, ECOG will not be pursuing additional trials of exisulind in NSCLC. PMID- 17409936 TI - Glutathione-S-transferase P1 isoenzyme polymorphisms, platinum-based chemotherapy, and non-small cell lung cancer. AB - BACKGROUND: Polymorphisms within the P1 isoenzyme of GST (GSTP1) are associated with alterations in enzyme activity and may change sensitivity to platinum-based chemotherapy. We investigated the relationship between exon 5 and exon 6 GSTP1 gene polymorphisms and treatment response, hematological, and nonhematological toxicity and overall survival for patients receiving platinum-based chemotherapy for advanced non-small cell lung cancer (NSCLC). METHODS: Between 2001 and 2002, 108 patients with chemotherapy-naive advanced NSCLC were recruited. Associations between the GSTP1 polymorphisms (Ile105Val, Thr110Ser, Ala114Val, and Asp 147Tyr) and GSTP1*A, *B, and *C haplotypes and treatment response and toxicity were evaluated using the Pearson chi and Kruskal-Wallis tests, respectively. Associations with survival were compared using Kaplan-Meier survival curves and Cox proportional hazard ratios. RESULTS: No significant associations were noted between GSTP1 polymorphisms and treatment response or survival. Significantly less neutropenic toxicity was demonstrated for patients possessing the 105Val allele (p = 0.020) or the GSTP1*B haplotype (p = 0.038). However, the variant allele GSTP1 105Val, and patients possessing a GSTP1*B allele demonstrated notable trends toward inferior response and survival. CONCLUSIONS: GSTP1 haplotype can be used to stratify hematological toxicity after platinum-based chemotherapy, but the lack of significant associations with response or survival suggests that GSTP1 polymorphisms may not be strong pharmacogenomic markers in this population. Additional large prospective studies incorporating the GSTP1 haplotype may clarify the reported discrepancies. PMID- 17409937 TI - Is the importance of achieving stable disease different between epidermal growth factor receptor tyrosine kinase inhibitors and cytotoxic agents in the second line setting for advanced non-small cell lung cancer? AB - BACKGROUND: It is controversial whether achieving stable disease leads to a survival benefit and whether the importance of achieving stable disease differs between cytotoxic agents and molecular targeted agents. To examine these questions, the authors retrospectively reviewed phase II and III studies in the second-line setting for advanced non-small cell lung cancer using epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) and cytotoxic agents separately. METHODS: The authors chose 45 trials for the chemotherapy group and nine for the EGFR TKI group by searching the PubMed database. All nine trials in the EGFR TKI group concern gefitinib and erlotinib. RESULTS: The median survival time increased 0.0375 month with each 1% increase in stable disease rate (p = 0.039), and each 1% increase in response rate resulted in 0.0744 (p < 0.001) month of median survival time in the analysis combined with both cytotoxic agents and EGFR TKIs. Main and interaction terms for EGFR TKI treatment were not statistically significant. With respect to time to progression, only response rate showed a statistically significant relationship with survival. CONCLUSIONS: To obtain response seems to be more important than to achieve stable disease for both cytotoxic agents and EGFR TKIs, although achieving stable disease is still valuable. The relationship between survival and response or stable disease appears similar for cytotoxic agents and EGFR TKIs. PMID- 17409938 TI - Impact of a multidisciplinary thoracic oncology clinic on the timeliness of care. AB - BACKGROUND: Multidisciplinary clinics have been recommended for the evaluation of patients with lung cancer. Evidence to support this recommendation, however, is limited. A single-center, retrospective review of lung cancer patients at a Veterans Affairs hospital was performed comparing timeliness of diagnostic and treatment decisions during the operation of a multidisciplinary thoracic oncology clinic (MTOC) with a period after it closed (non-MTOC), during which only a weekly multidisciplinary conference was held. METHODS: Patients were identified from a tumor registry. Manual chart reviews were performed on all patients. Outcome measures included time from initial presentation to diagnosis (TTD) and time from diagnosis to treatment initiation (TTT). RESULTS: Three hundred forty five patients (244 in MTOC, 101 in non-MTOC) diagnosed with lung cancer between 1999 and 2003 were included in the study. Baseline characteristics were similar between the two groups. Median TTD was 48 days (95% confidence interval [CI]: 37 61) and 47 days (95% CI: 39-55) in the MTOC (n = 164) and non-MTOC cohorts (n = 89), respectively (p = 0.09). Median TTT was 22 days (95% CI: 20-27) and 23 days (95% CI: 20-34) in the MTOC (n = 165) and non-MTOC cohorts (n = 89), respectively (p = 0.71). There was no difference in overall survival. CONCLUSION: Retrospective comparison of sequential cohorts failed to reveal benefit in the timeliness of care measures during the time period of MTOC operation. Potential confounders include the absence of a surgeon in the MTOC setting, an ongoing weekly multidisciplinary conference in the non-MTOC cohort, and existing infrastructures based on previous MTOC experiences and past provider experience. Confirmation of these findings in other health care settings is warranted, preferably in a prospective fashion. PMID- 17409939 TI - A simple solution for complicated pleural effusions. AB - INTRODUCTION: Complicated pleural effusions are difficult to manage with conventional strategies. In this study, we review the safety, efficacy, and durability of PleurX catheters (Denver Biomedical, Golden, CO) for managing complicated pleural effusions and describe a patient population who might benefit. PATIENTS AND METHODS: From July 1999 to February 2003, 63 PleurX catheters were placed in 58 symptomatic patients (an additional five had bilateral catheters) to manage complicated pleural effusions. Patients selected for catheter placement tended to have poor performance status (Eastern Cooperative Oncology Group < or =2) or had failed standard therapies. Of the 63 catheters, 52 (83%) were placed because of malignant complicated pleural effusions. A registry of patients was constructed, and data were obtained from review of medical records. Nonparametric estimates of freedom from reintervention and overall survival were obtained by the Kaplan-Meier method. RESULTS: Catheter related complications were noted in four of 58 patients (7%) and included one each of pneumothorax, seroma, empyema, and pain syndrome. Freedom from reintervention for effusion management was 95%. Of the patients, 86% (50 of 58) experienced dyspnea relief. There were no procedure-related mortalities. Catheters remained functional up to 330 days, and four of 63 (6%) required one time thrombolysis with tissue plasminogen activator. CONCLUSIONS: PleurX catheters are safe, effective, and durable solutions for complicated pleural effusions and seem to provide an attractive alternative for patients who have few other palliative options. We consider the catheters as first-line therapy for these patients. PMID- 17409941 TI - Bilateral choroidal metastases from mesothelioma. PMID- 17409940 TI - Targeting mesothelioma using an infectivity enhanced survivin-conditionally replicative adenoviruses. AB - Mesothelioma is a highly malignant neoplasm with no effective treatment. Conditionally replicative adenoviruses (CRAds) represent a promising new modality for the treatment of cancer in general. A key contribution in this regard is the introduction of tumor-selective viral replication for amplification of the initial inoculum in the neoplastic cell population. Under ideal conditions following cellular infection, the viruses replicate selectively in the infected tumor cells and kill the cells by cytolysis, leaving normal cells unaffected. However, to date there have been two limitations to clinical application of these CRAd agents; viral infectivity and tumor specificity have been poor. Herein we report on two CRAd agents, CRAd-S.RGD and CRAd-S.F5/3, in which the tumor specificity is regulated by a tumor-specific promoter, the survivin promoter, and the viral infectivity is enhanced by incorporating a capsid modification (RGD or F5/3) in the adenovirus fiber region. These CRAd agents effectively target human mesothelioma cell lines, induce strong cytoxicity in these cells in vitro, and viral replication in a H226 murine xenograft model in vivo. In addition, the survivin promoter has extremely low activity both in the non-transformed cell line, HMEC, and in human liver tissue. Our results suggest that the survivin based CRAds are promising agents for targeting mesothelioma with low host toxicity. These agents should provide important insights into the identification of novel therapeutic strategies for mesothelioma. PMID- 17409942 TI - Choroidal metastasis of small cell lung cancer. PMID- 17409943 TI - Targeting anemia in patients with lung cancer. AB - Anemia is highly prevalent in patients with lung cancer, often occurring at baseline and frequently exacerbated as a result of treatment with platinum-based chemotherapy. Anemia has been shown to have a negative effect on quality of life in patients with lung cancer, and additional data indicate that decreases in hemoglobin in these patients are associated with impaired survival. Multiple clinical studies have demonstrated that treatment of anemia with erythropoietic agents in patients with lung cancer results in a significant increase in hemoglobin, decrease in transfusions, and improvement in quality of life. Ongoing research is evaluating whether erythropoietic therapy can reduce cognitive impairment associated with lung cancer, cytotoxic therapy, and anemia. Despite the known adverse effects of anemia and the established benefits of erythropoietic therapy in anemic patients with lung cancer, more than half of these patients do not receive any anemia treatment. The purpose of this review is to report results of the European Cancer Anaemia Survey that describe the prevalence of anemia in patients with lung cancer, to review the major studies evaluating the clinical outcomes of erythropoietic therapy in patients with lung cancer, to discuss the recent safety concerns regarding the use of erythropoietic agents in patients with cancer treated to high hemoglobin levels, and to describe various novel therapeutic applications of erythropoietic agents in lung cancer. PMID- 17409944 TI - Inflammatory myofibroblastic tumor of the left main stem bronchus. AB - Inflammatory myofibroblastic tumor (IMT) (pseudotumor) is a rare solid tumor that most commonly affects the lung. Its unknown etiology and cellular variability have resulted in inconsistent naming of this lesion. Treatment modalities have varied, but complete surgical resection seems to be most appropriate. We describe a 28-year-old woman with a near-obstructing IMT of the left main stem bronchus (LMB) treated with bronchial sleeve resection. PMID- 17409945 TI - Mediastinal and retroperitoneal teratoma with focal gastrointestinal adenocarcinoma. AB - We report an unusual case of gastrointestinal adenocarcinoma arising in a giant posterior mediastinal mature cystic teratoma extending into the retroperitoneum, which was treated by complete excision with a good outcome for more than 2 years. Teratomas with malignant transformation are rare non-germ cell malignant tumors arising from a preexisting mature teratoma. Histological examination revealed that the cyst wall was composed of mature ectodermal, mesodermal, and endodermal elements. Neoplastic glands with a cribriform pattern were found in a small, solid nodule. Strong cytokeratin 20 cytoplasmic immunostaining of the tumor cells supported the diagnosis of gastrointestinal adenocarcinoma. In this report, we describe the potential aggressiveness of a giant mature cystic teratoma with adenocarcinoma and suggest that complete surgical resection without adjuvant chemotherapy be considered as a therapy in the treatment of teratoma with focal malignant transformation. PMID- 17409946 TI - Temozolomide as prophylaxis for brain metastasis in non-small cell lung cancer. PMID- 17409947 TI - Topical retinoids: another piece for the retinoid-cigarette-lung cancer puzzle? PMID- 17409949 TI - Efficacy of gefitinib in a patient with lung cancer associated with idiopathic pulmonary fibrosis. PMID- 17409950 TI - Will we ever have personalized medicine for non-small cell lung cancer? PMID- 17409951 TI - Update on epidermal growth factor receptor inhibitor development in lung cancer. AB - Therapeutic strategies targeting the epidermal growth factor receptor (EGFR) are being evaluated in a number of ongoing clinical studies in non-small cell lung cancer (NSCLC). EGFR tyrosine kinase inhibitors (TKIs) are currently used worldwide to treat advanced refractory NSCLC, whereas the efficacy of anti-EGFR monoclonal antibodies remains to be established. Using molecular profiling to select patients to receive earlier EGFR targeted therapy as first-line treatment or in the adjuvant setting is an area of active research. Based on preclinical data, combining EGFR TKIs with other treatment modalities has promise, and clinical validation is underway. A new generation of irreversibly bound EGFR TKIs is being developed, and insights into the molecular biology of NSCLC should help to better define the patients who are most likely to benefit from these compounds. We summarize updates on EGFR targeted therapies that were presented during the sixth annual Targeted Therapies for the Treatment of Lung Cancer Conference in Los Angeles, CA; January 27-28, 2006. PMID- 17409952 TI - Angiogenesis inhibitors for lung cancer: clinical developments and future directions. PMID- 17409953 TI - The Akt/mTOR and mitogen-activated protein kinase pathways in lung cancer therapy. AB - Aberrant intracellular signaling resulting from mutations and oncogenic activation, as well as gene amplification of critical proteins involved in signal transduction pathways, are key features of lung cancer. Three important intracellular signaling proteins, the mammalian target of rapamycin, protein kinase B, and mitogen-activated protein kinase kinase have emerged as attractive targets for lung cancer therapy. We review current information on the therapeutic manipulation of these targets and describe early clinical data in lung cancer. PMID- 17409954 TI - Novel cytotoxic agents for non-small cell lung cancer. AB - Cytotoxic agents have unequivocal activity in non-small cell lung cancer. Currently available agents have demonstrated the ability to prolong life and improve quality of life in advanced disease, and to increase the rate of cure when used in stage I and II disease in the adjuvant setting and as part of chemoradiotherapy in stage III disease. These agents have served as the core regimen to which agents effecting newly discovered molecular targets are added. However, there is little question that there is much room for improvement. A number of new agents have been identified that either derive from older drugs or affect their targets in novel ways. In particular, tubulin has been an attractive target since the dawn of medical oncology. A number of new antitubulin agents are currently in development. Epothilones are novel agents that bind to the same site as taxanes, yet are structurally distinct. Xyotax and abraxane are agents in which paclitaxel is delivered in a novel fashion that may both reduce toxicity and enhance activity. ABT 751 is an agent that targets the colchicine site of tubulin. Another approach is to enhance the activity of currently available agents by targeting detoxification and resistance pathways. TLK-286 is a novel agent that may enhance the activity of previously available agents by inhibiting GST-pi, a detoxifying mechanism that may be of particular relevance to platinum agents. It has also demonstrated some single-agent cytotoxic activity. PMID- 17409955 TI - Immune-modulating vaccines in non-small cell lung cancer. AB - The limited efficacy and moderate toxicity of chemotherapy for non-small cell lung cancer (NSCLC) have given the search for a more effective and safer systemic treatment a sense of urgency. NSCLC has historically been regarded as a non immunogenic cancer, and immunotherapy has yielded little benefit in humans. However, encouraging clinical results in patients immunized with novel vaccines directed toward enhancement of tumor antigen recognition have been published. In this review, we describe the scientific basis and updates the clinical results of three vaccine approaches (GVAX, Lucanix, and L-BLP25) and discuss preliminary work on a novel Ras mutation targeted immune therapy. Further studies are needed to demonstrate whether these novel therapies can potentially complement or replace current therapeutic approaches. PMID- 17409956 TI - Definitive radiotherapy for stage I non-small cell lung cancer: ain't nothing like the real thing? PMID- 17409957 TI - Treatment of choice for stage I non-small cell lung cancer: surgery or radiotherapy? PMID- 17409958 TI - Vascular endothelial growth factor (VEGF) pathway. PMID- 17409959 TI - Integrin-linked kinase inhibitor KP-392 demonstrates clinical benefits in an orthotopic human non-small cell lung cancer model. AB - INTRODUCTION: The overexpression of integrin-linked kinase (ILK) has been implicated in the promotion of tumor invasion and metastasis. We studied the anticancer effects of KP-392, a potent selective inhibitor of ILK in the NCI-H460 cell line. In vitro, KP-392 inhibited ILK activity of H460 cells. In vivo, the effect of KP-392 was investigated in a metastatic H460 orthotopic lung cancer model. METHODS: Intraperitoneal KP-392 (5 mg/day per animal) was administered both alone and in combination with cisplatin (5 mg/kg per week for 3 weeks). In group I, all treated animals were followed until death to assess therapeutic effect on survival. In group II, tumor growth and metastasis were evaluated by sacrificing one animal from each treatment when a control animal died. RESULTS: Both cisplatin and KP-392 significantly enhanced survival (37.8 +/- 3.7 and 34.9 +/- 5.2 days) compared with the control (30.2 +/- 3.6 days, p < 0.0001 and p = 0.0418, respectively), and the survival benefit from combination treatment was greater than that of either agent alone (45.8 +/- 3.9 days, p < 0.0001). Although KP-392 alone did not impact the incidence of metastasis, in combination with cisplatin a consistent trend of inhibition was seen for metastases in the kidney, bone, and the contralateral lung. KP-392 was well tolerated throughout the study. KP-392 demonstrated increased tumor necrosis and decreased nuclear phospho protein kinase/Akt but did not change the levels of phospho-extracellular signal regulated kinase 1/2. CONCLUSIONS: ILK inhibitor does not enhance the toxicity of standard chemotherapy and may have a beneficial therapeutic effect in lung cancer. PMID- 17409960 TI - Analysis of differentially expressed genes in neuroendocrine carcinomas of the lung. AB - INTRODUCTION: Large cell neuroendocrine carcinoma (LCNEC) and small cell lung carcinoma (SCLC) show considerable differences in their histology but share neuroendocrine (NE) characteristics and also genetic and/or expression patterns. METHODS: We used the subtractive expression method to identify differences in gene expression that would allow discrimination between these two types of NE lung carcinoma. RESULTS: Eight cDNA fragments were transcribed at a higher level in LCNEC compared with SCLC, and these corresponded to five mitochondrial genes, two ribosomal genes, and one fetal regulation factor, neuronatin (NNAT). Immunohistochemically, NNAT protein was detected in 43% (6/14) of LCNECs but in only 8% (1/13) of SCLCs (p < 0.05). Positive staining for NNAT was observed in areas that did not show the NE morphology, such as palisading and rosettes. CONCLUSIONS: The present results suggest that NNAT has the potential to be used as a differential maker between LCNEC and SCLC. PMID- 17409961 TI - Clinico-pathological and biological significance of tyrosine kinase domain gene mutations and overexpression of epidermal growth factor receptor for lung adenocarcinoma. AB - INTRODUCTION: Mutations in the tyrosine kinase domain (TKD) of the epidermal growth factor receptor (EGFR) gene have proven to be clinically significant in non-small cell lung cancer. However, relationships between these mutations and EGFR expression or deletion mutations in the extracellular domain of EGFR (EGFRvIII) remain unclear. The purpose of this study was to gain further insight into the clinical significance of these molecular abnormalities in lung adenocarcinoma. METHODS: We investigated EGFR TKD mutations using direct sequencing, EGFR protein expression using Western blotting, and EGFRvIII using reverse transcriptase-polymerase chain reaction in samples from 48 adenocarcinoma patients. Correlations with various clinico-pathological features were analyzed. RESULTS: EGFR TKD mutations were detected in 25 of 48 adenocarcinomas (52.1%), and overexpression of EGFR protein was identified in 19 patients (39.6%). Presence of EGFR TKD mutations was significantly correlated with EGFR overexpression (p = 0.021). EGFR TKD mutations were significantly correlated with never-smoker status (p = 0.043), absence of emphysematous or fibrotic appearance on computed tomography (p = 0.001), papillary subtype (p = 0.041), and bronchioloalveolar carcinoma features (p = 0.045). EGFRvIII was not detected in any adenocarcinomas. Retrospective analysis revealed that patients with EGFR TKD mutations displayed better postoperative prognosis than patients with wild-type EGFR (p = 0.033). CONCLUSIONS: These results suggest that EGFR TKD mutation is associated with EGFR overexpression, representing an important factor for consideration when investigating the clinical significance, including susceptibility to chemotherapy, of EGFR TKD mutations in adenocarcinoma. EGFRvIII does not seem to play a major role in the development of lung adenocarcinoma. PMID- 17409962 TI - Stage I non-small cell lung cancer: results for surgery in a patterns-of-care study in Sydney and for high-dose concurrent end-phase boost accelerated radiotherapy. AB - PURPOSE: To review results of resection or radical radiotherapy in Sydney to inform patients contemplating treatment options for early-stage non-small cell lung cancer (NSCLC). METHODS AND MATERIALS: Sixty-eight resected pathologic stage I patients were identified from a patterns-of-care (POC) study of all 1993 and 1996 NSCLC patients treated at the South Western Sydney Area Health Service and the Northern Sydney Area Health Service. These were compared with 39 clinical stage I patients treated by modern radiotherapy using an accelerated high-dose technique at St George Hospital (SGH). RESULTS: Resected POC patients achieved a median survival rate of 67 months and a 5-year survival rate of 54% (95% CI, 40 66%). SGH radiotherapy patients achieved a median survival rate of 43 months and a 5-year survival rate of 30% (95% CI, 13-48%). On multivariate analysis, the only significant factor is performance status, which may reflect comorbidity burden. Cancer-specific 5-year survival rates for surgery (59% [44-71%]) and for radiotherapy (53% [28-72%]) are not significantly different. CONCLUSIONS: Modern radiotherapy is a reasonable option for patients to consider for stage I NSCLC and is recommended for medically inoperable patients rather than supportive care or observation. PMID- 17409963 TI - Results of a phase I dose-escalation study using single-fraction stereotactic radiotherapy for lung tumors. AB - BACKGROUND: The purpose of this study was to report initial results of a phase I study using single-fraction stereotactic radiotherapy (RT) in patients with inoperable lung tumors. METHODS: Eligible patients included those with inoperable T1-2N0 non-small cell lung cancer (NSCLC) or solitary lung metastases. Treatments were delivered by means of the CyberKnife. All patients underwent computed tomography-guided metallic fiducial placement in the tumor for image-guided targeting. Nine to 20 patients were treated per dose cohort starting at 15 Gy/fraction followed by dose escalation of 5 to 10 Gy to a maximal dose of 30 Gy/fraction. A minimal 3-month period was required between each dose level to monitor toxicity. RESULTS: Thirty-two patients (21 NSCLC and 11 metastatic tumors) were enrolled. At 25 Gy, pulmonary toxicity was noted in patients with prior pulmonary RT and treatment volumes greater than 50 cc; therefore, dose escalation to 30 Gy was applied only to unirradiated patients and treatment volume less than 50 cc. Ten patients received doses less than 20 Gy, 20 received 25 Gy, and two received 30 Gy. RT-related complications were noted for doses greater than 25 Gy and included four cases of grade 2 to 3 pneumonitis, one pleural effusion, and three possible treatment-related deaths. The 1-year freedom from local progression was 91% for dose greater than 20 Gy and 54% for dose less than 20 Gy in NSCLC (p = 0.03). NSCLC patients had significantly better freedom from relapse (p = 0.003) and borderline higher survival than those with metastatic tumors (p = 0.07). CONCLUSIONS: Single-fraction stereotactic RT is feasible for selected patients with lung tumors. For those with prior thoracic RT, 25 Gy may be too toxic. Higher dose was associated with improved local control. Longer follow-up is necessary to determine the treatment efficacy and toxicity. PMID- 17409964 TI - Docetaxel consolidation therapy following cisplatin, vinorelbine, and concurrent thoracic radiotherapy in patients with unresectable stage III non-small cell lung cancer. AB - BACKGROUND: To evaluate the feasibility and efficacy of docetaxel consolidation therapy after concurrent chemoradiotherapy for unresectable stage III non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: The eligibility criteria included unresectable stage III NSCLC, no previous treatment, age between 20 and 74 years, and performance status 0 or 1. Treatment consisted of cisplatin (80 mg/m2 on days 1, 29, and 57), vinorelbine (20 mg/m2 on days 1, 8, 29, 36, 57, and 64), and thoracic radiotherapy (TRT) (60 Gy/30 fractions over 6 weeks starting on day 2), followed by consolidation docetaxel (60 mg/m2 every 3 to 4 weeks for three cycles). RESULTS: Of 97 patients who were enrolled in this study between 2001 and 2003, 93 (76 males and 17 females with a median age of 60) could be evaluated. Chemoradiotherapy was well tolerated; three cycles of chemotherapy and 60 Gy of TRT were administered in 80 (86%) and 87 (94%) patients, respectively. Grade 3 or 4 neutropenia, esophagitis, and pneumonitis developed in 62, 11, and 3 patients, respectively. Docetaxel consolidation was administered in 59 (63%) patients, but three cycles were completed in only 34 (37%) patients. The most common reason for discontinuation was pneumonitis, which developed in 14 (24%) of the 59 patients. During consolidation therapy, grade 3 or 4 neutropenia, esophagitis, and pneumonitis developed in 51, 2, and 4 patients, respectively. A total of four patients died of pneumonitis. We calculated a V20 (the percent volume of the normal lung receiving 20 Gy or more) on a dose-volume histogram in 25 patients. Of these, five patients developed grade 3 or more severe radiation pneumonitis. A median V20 for these five patients was 35% (range, 26-40%), whereas the median V20 for the remaining 20 patients was 30% (range, 17-35%) (p = 0.035 by a Mann Whitney test). The response rate was 81.7% (95% confidence interval [CI], 72.7 88.0%), with 5 complete and 71 partial responses. The median progression-free survival was 12.8 (CI, 10.2-15.4) months, and median survival was 30.4 (CI, 24.5 36.3) months. The 1-, 2-, and 3-year survival rates were 80.7, 60.2, and 42.6%, respectively. CONCLUSION: This regimen produced promising overall survival in patients with stage III NSCLC, but the vast majority of patients could not continue with the docetaxel consolidation because of toxicity. PMID- 17409965 TI - Palliative thoracic radiotherapy in locally advanced non-small cell lung cancer: can quality-of-life assessments help in selection of patients for short- or long course radiotherapy? AB - PURPOSE: Patient-assessed health-related quality-of-life (HRQOL) scores, together with demographic and clinical factors in stage III non-small cell lung cancer (NSCLC) patients, are important prognostic factors for survival and may be helpful in determining thoracic radiotherapy (TRT) strategy. METHODS: In a previously published randomized trial, 301 patients were treated with different palliative radiotherapy schedules, comparing short-term hypofractionated TRT (arm A: 17 Gy/2 fractions [n = 105]) with more protracted TRT (arm B: 42 Gy/15 fractions [n = 104]); arm C: 50 Gy/25 fractions [n = 92]). Baseline HRQOL, demographic, and clinical data were available for all patients. All possible prognostic factors from univariate analysis were entered into the Cox multivariate regression model to identify variables of independent prognostic relevance. RESULTS: Overall survival was similar, whereas long-term survival was restricted to higher-dose radiotherapy with 3-year survival rates of 1, 8, and 6% (p = 0.40) and 5-year survival rates of 0, 4, and 3% (p = 0.12) in arms A, B, and C, respectively. In univariate analysis, Karnofsky performance status, use of analgesics, and weight loss were highly significant non-HRQOL factors (p < 0.001), and physical function, appetite loss, cough, and pain were the most powerful HRQOL factors (p < 0.001). In multivariate analysis, appetite loss appeared as the most powerful independent prognostic indicator. In the group of patients treated with protracted fractionation (n = 196), the 2-, 3-, and 5-year survival rates in patients with no appetite loss (n = 95) were 22% (21/95), 12% (11/95), and 8% (8/95) compared with 3% (3/101), 1% (1/101), and 1% (1/101) in patients with appetite loss present at baseline (n = 101). CONCLUSION: In addition to performance status and weight loss, patient-reported appetite loss should be assessed in stage III NSCLC patients before administrating TRT; such assessment is a valuable tool for selecting patients to normofractionated or lower-dose hypofractionated palliative TRT. PMID- 17409966 TI - Clinical characterization of node-negative lung adenocarcinoma: results of a prospective investigation. AB - OBJECTIVES: In clinical stage IA lung adenocarcinoma, the clinical features of a truly node-negative population were prospectively investigated by means of a prospective investigational study. METHODS: The clinical data and radiographic features of high-resolution computed tomography (HR-CT) were registered before operation in 169 clinical stage IA lung cancer patients who were scheduled to undergo a standard lobectomy and systemic mediastinal node dissection. The nodal metastasis was pathologically defined and the clinical factors associated with the presence of the nodal metastasis were evaluated. RESULTS: In 114 assessable cases with adenocarcinomas, 15 (13.1%) were node-positive. The serum carcinoembryonic antigen (CEA), retraction sign, and intratumoral air-bronchogram on HR-CT were suggested to be predictive factors for lymph node metastasis, with hazard ratios of 12.44 (p = 0.0003), 6.53 (p = 0.0533), and 0.17 (p = 0.0073), respectively. In combination with the radiologic features and serum CEA, cases with elevated serum CEA or presence of retraction sign included 15.6% of node metastasis-positive, whereas all cases with normal CEA and absence of retraction sign showed no nodal metastasis. Cases with elevated serum CEA or absence of intratumoral air-bronchogram included 24.5% of node metastasis, whereas cases with normal CEA and presence of air-bronchogram showed 4.6% of node metastasis. The tumor size and the proportion of ground-glass attenuation were not associated with the incidence of nodal metastasis. CONCLUSIONS: The serum CEA and HR-CT features thus allowed us to identify node-negative lung adenocarcinomas measuring 3 cm or less in size. PMID- 17409967 TI - Phase II study of carboplatin, irinotecan, and thalidomide in patients with advanced non-small cell lung cancer. AB - BACKGROUND: We hypothesized that thalidomide would improve the response and toxicity profile of chemotherapy with carboplatin and irinotecan. METHODS: The key eligibility criteria were stage IIIB (malignant pleural effusion) and IV non small cell lung cancer, measurable disease, no prior chemotherapy, prior radiation only for brain metastasis, performance status 0 or 1, and adequate hematologic, hepatic, and renal function. Treatment consisted of carboplatin at a calculated area under the curve of 5 and infused intravenously for 30 min on day 1 and irinotecan (50 mg/m2 intravenously for 90 min on days 1 and 8 every 21 days). Thalidomide was given orally every evening starting on day 1 until progressive disease; the starting dose was 200 mg per day, which was escalated by 100 mg per week if tolerated (maximum 1000 mg per day). The objectives were to determine the response rate, time to progression, overall survival, and toxicity profile. RESULTS: In all, 46 patients were enrolled, but three who never received protocol treatment were excluded from the analysis. The characteristics of the 43 eligible and treated patients included median age 63 (47-79), female/male 13/30, black/white 3/40, PS 0/1 in 10/33, and stage 3/4 in 6/37. The objective response rates were complete response 1 (2%), partial response 5 (12%), stable disease 24 (56%), progressive disease 9 (21%), and unevaluable for response 4 (9%). The median time to progression was 3.7 months (95% confidence interval [CI], 2.5 4.9). The median survival time was 8.1 months (95% CI, 5.0-12.9). Frequent toxicities were neutropenia, fatigue/malaise, and nausea/vomiting. Diarrhea was uncommon and mild. CONCLUSIONS: This treatment regimen of carboplatin, irinotecan, and thalidomide was tolerable, with reversible neutropenia as the major toxicity and only minor diarrhea. The overall response rate did not meet our predetermined level of efficacy to merit further investigation. PMID- 17409968 TI - Clinical utility of epidermal growth factor receptor expression for selecting patients with advanced non-small cell lung cancer for treatment with erlotinib. AB - INTRODUCTION: Erlotinib (Tarceva) has demonstrated a survival benefit in unselected patients with advanced non-small cell lung cancer (NSCLC) after failure of chemotherapy. Because not all patients benefit from erlotinib, epidermal growth factor receptor (EGFR) protein expression may provide a basis for selecting patients for treatment with this EGFR inhibitor. METHODS: Tumor samples from patients who participated in National Institute of Canada Clinical Trials Group Study BR.21 were assayed by immunohistochemistry using Dako EGFR pharmDx kits. EGFR expression was scored as proportion of tumor cells with membrane staining, staining intensity, and combined proportion and intensity scores. All possible cutpoints were examined to determine whether EGFR protein expression status by immunohistochemistry might be useful for predicting patient survival. RESULTS: Three hundred twenty-five patients had evaluable assay results. The prognostic significance of EGFR protein expression was modest. Patients with EGFR-positive tumors who received placebo after failure of chemotherapy had slightly worse survival than patients with EGFR-negative tumors; however, the differences were not statistically significant for any cutpoint for any of the three measures of EGFR protein expression. The treatment benefits from erlotinib relative to placebo were greater for EGFR-positive patients compared with EGFR-negative patients, but they were not significantly different for any cutoff used to define EGFR positivity. Use of very high cutpoints to define patients with EGFR-rich tumors that might be especially sensitive to treatment with erlotinib cannot be supported by these data. CONCLUSIONS: Selection or exclusion of NSCLC patients for erlotinib therapy after failure of standard therapy for advanced or metastatic disease should not be based solely on EGFR protein expression as determined by immunohistochemistry. PMID- 17409969 TI - Gefitinib (IRESSA) in patients of Asian origin with refractory advanced non-small cell lung cancer: subset analysis from the ISEL study. AB - INTRODUCTION: The IRESSA Survival Evaluation in Lung Cancer (ISEL) phase III study compared the efficacy of gefitinib (IRESSA) versus placebo in patients with refractory advanced non-small cell lung cancer (NSCLC). Although a statistically significant difference in survival was not seen between gefitinib and placebo in the overall ISEL population, preplanned subset analyses demonstrated a significant survival benefit in patients who had never smoked and in patients of Asian origin. METHODS: In ISEL, 1692 patients who were refractory to or intolerant of their latest chemotherapy were randomized to receive either gefitinib (250 mg/day) or placebo, plus best supportive care. Preplanned subgroup analyses included an assessment of patients who were of Asian origin (n = 342). RESULTS: Two hundred thirty-five patients of Asian origin received gefitinib, and 107 received placebo. In these patients, treatment with gefitinib significantly improved survival compared with placebo (hazard ratio [HR], 0.66; 95% confidence interval [CI], 0.48, 0.91; p = 0.010; median survival, 9.5 versus 5.5 months). Patients of Asian origin also experienced statistically significant improvements in time to treatment failure with gefitinib compared with placebo (HR, 0.69; 95% CI, 0.52, 0.91; p = 0.0084; 4.4 versus 2.2 months), and objective response rates were higher with gefitinib than with placebo (12 versus 2%). Gefitinib was generally well tolerated in patients of Asian origin, with rash and diarrhea being the most common adverse events. No unexpected adverse events were observed. CONCLUSIONS: Treatment with gefitinib was associated with a significant improvement in survival in a subgroup of patients of Asian origin with previously treated refractory advanced NSCLC. PMID- 17409970 TI - Fulminant thoracic sarcoma masked by hemothorax. PMID- 17409971 TI - Asymptomatic endobronchial metastases of typical carcinoid 15 years after curative resection. PMID- 17409972 TI - Increasing osteoblastic lesions as a manifestation of a major response to gefitinib. PMID- 17409973 TI - Second-line treatment of non-small cell lung cancer: big targets, small progress; small targets, big progress? PMID- 17409974 TI - MicroRNA in lung cancer. PMID- 17409975 TI - Establishment and expression profiling of new lung cancer cell lines from Chinese smokers and lifetime never-smokers. AB - BACKGROUND: Bronchogenic adenocarcinoma is the predominant histologic subtype of lung cancer, which ranks top in the cancer mortality in both men and women. Female nonsmokers and adenocarcinoma have emerged as a distinct combination in patients with lung cancer in recent decades. Lung cancer cell lines established from patients with known clinical characteristics such as gender and smoking habit would be useful for future research on lung cancer. METHODS: Four new lung adenocarcinoma cell lines (HKULC 1-4) were established from Chinese patients with primary lung adenocarcinomas and with different clinical characteristics with respect to age, gender, smoking habits, tumor staging, and previous therapy. They were characterized by immunohistochemical and growth kinetic studies, tests for tumorigenicity in nude mice, epidermal growth factor receptor (EGFR) gene mutation analysis, and in situ hybridization, and gene expression profiling with Affymetrix GeneChip HG-U133A. RESULTS: The newly established HKULC lung adenocarcinoma cell lines were maintained for over 70 passages and demonstrated morphologic and immunohistochemical features and growth kinetics of tumor cell lines. One of the four HKULC cell lines, HKULC 3 (derived from a female nonsmoking patient with lung adenocarcinoma), was found to have a deletion at exon 19 of the EGFR gene. EGFR in situ hybridization showed no EGFR gene amplification in these cell lines. HKULC 1 and 4 formed tumor xenografts after inoculation in nude mice. A list of 71 genes that were differentially expressed or showing class predictive significance was identified. These genes included putative tumor suppressor genes (DKK3, SERPINF1, CDH11, DSC3, and KLF6), genes involved in or related to the EGFR pathways (ERBB3, MUC1, VAV1), genes involved in regulation of cell cycle and proliferation (CDKN1A and CDKN2A), a putative oncogene (EEF1A2), and a gene related to metastasis (MTSS1). DISCUSSION: Four new lung adenocarcinoma cell lines were established from patients with different clinical characteristics. These characterized cell lines and their gene expression profiles will provide resources for studies of lung cancer biology and in vitro chemotherapeutic drug study. PMID- 17409976 TI - Misclassification of bronchioloalveolar carcinoma with cytologic diagnosis of lung cancer. AB - INTRODUCTION: Cytology is commonly used to diagnose non-small cell lung cancer (NSCLC) but is an inaccurate means of diagnosis of bronchioloalveolar carcinoma (BAC). The aims of this study were to calculate the sensitivity and specificity of cytologic diagnosis of BAC and to estimate the misclassification of BAC as other subtypes of NSCLC. METHODS: Preoperative fine-needle aspiration cytology diagnoses were compared to histology diagnoses in 222 patients, including 51 patients with pure or mixed BAC, who underwent lung resection for NSCLC at our institution since 1999. RESULTS: The sensitivity and specificity of a cytologic diagnosis of BAC were 12% and 99%, respectively. Based on cytologic diagnosis, 63% of BAC was misclassified as adenocarcinoma, and 18% was misclassified as undifferentiated NSCLC. In this cohort, 35% of adenocarcinomas and 12% of undifferentiated NSCLC diagnosed by cytology had BAC histology. CONCLUSIONS: Diagnosis of NSCLC by cytology alone results in significant misclassification of BAC, most commonly as adenocarcinoma or undifferentiated NSCLC. Because patients with BAC respond differently to certain treatments such as endothelial growth factor receptor inhibitors and surgical resection of multifocal lung cancer, misclassification of BAC may have important therapeutic implications. PMID- 17409977 TI - Size of metastatic and nonmetastatic mediastinal lymph nodes in non-small cell lung cancer. AB - OBJECTIVE: To determine the optimum selection of mediastinal lymph nodes for biopsy in non-small cell lung cancer (NSCLC), lymph nodes with or without metastasis at each mediastinal station were ranked in size in patients with pathological N2 disease. METHODS: Twenty-five NSCLC patients with pathological N2 disease who underwent pulmonary resection with complete mediastinal lymph node clearance were examined. Of 114 mediastinal lymph node stations dissected, 47 had metastases and 67 did not. The sizes of 259 nodes in the 47 positive lymph node stations were measured. Of these 259 nodes, 137 had metastases and 122 did not. The short- and long-axis diameters of the 259 lymph nodes were ranked in each lymph node station. RESULTS: Mean short- and long-axis diameters of lymph nodes with metastases were significantly greater than those without (p < 0.001). In 47 metastatic lymph node stations, the short- and long-axis diameters were greatest in a metastatic node in 44 (94%) and 42 (89%) respectively, whereas in the remaining 3 (6%) and 5 (11%), the second largest but not the largest node was positive. None of the largest lymph nodes with metastasis were smaller than the second largest lymph node at each station. Four of the 10 patients with adenocarcinoma (40%) had metastasis in the second largest but not in the largest node measured by long-axis diameter, a significant difference from one in eight (12.5%) among the squamous cell carcinoma cases (p = 0.04). CONCLUSION: For mediastinal lymph node biopsy, both the largest and the second largest node at each station should be sampled, especially in adenocarcinoma. If only the largest lymph node is selected, false-negative results will occur at a rate of about 10%. PMID- 17409978 TI - A clinical prediction rule to estimate the probability of mediastinal metastasis in patients with non-small cell lung cancer. AB - INTRODUCTION: Estimating the clinical probability of mediastinal metastasis in patients with non-small cell lung cancer (NSCLC) can facilitate the selection and interpretation of staging tests. PURPOSE: We sought to identify independent clinical predictors of mediastinal metastasis and to develop a prediction rule to estimate the pretest probability of nodal metastasis in patients with NSCLC. METHODS: We used data from a randomized controlled trial of selective versus routine mediastinoscopy to develop a clinical prediction model. RESULTS: Five hundred sixty-six patients were included, with a mean age of 65 +/- 9 years; 31% had positive lymph nodes. Independent predictors of positive nodes included adenocarcinoma or large cell histology (OR 2.6, 95% confidence interval [CI] = 1.8-3.9), apparent metastatic disease on chest radiography (OR 2.4, 95% CI = 1.2 4.7), central location of the primary tumor (OR 2.1, 95% CI = 1.4-3.3), symptoms from the primary tumor (OR 1.6, 95% CI = 1.1-2.4), tumor diameter > or =3.6 cm (OR 1.5, 95% CI = 1.0-2.3), and age less than 65 years (OR 1.5, 95% CI = 1.0 2.2). Model accuracy and calibration were good, with an area under the receiver operating characteristic curve of 0.70 (95% CI = 0.66-0.75) and good agreement between observed and predicted probabilities of mediastinal metastasis. CONCLUSIONS: Our prediction rule can be used to estimate the pretest probability of mediastinal metastasis in patients with NSCLC. Such estimates can facilitate clinical decision making when selecting and interpreting the results of noninvasive and invasive staging tests. PMID- 17409979 TI - Wedge resection for non-small cell lung cancer in patients with pulmonary insufficiency: prospective ten-year survival. AB - BACKGROUND: Possibility of curative resection by lobectomy for non-small cell lung cancer is often denied patients with compromised pulmonary reserve. Analysis of survival of such patients treated by wedge resection was compared with that of patients treated by standard resection, with both groups followed for 10 years. DESIGN: A prospective 5-year cohort study. METHODS: From 1988 to 1992, an observational cohort of 127 consecutive resected patients at Memphis VA Medical Center was divided into those receiving lobectomy in 81 cases and pneumonectomy in 15 cases (group I) versus 31 patients with compromised pulmonary reserve (group II), who had complete tumor excision by wedge resection. Preoperative clinical staging was corrected to surgical-pathological staging after demonstration of its superiority. Survival estimates were obtained by Kaplan Meier method with curves compared by log rank tests, with all-cause mortality calculated from date of surgery. RESULTS: Extent of disease in group I was 58% stage I, 19% stage II, and 23% stage III. In group II, extent of disease was 84% stage I, 3% stage II, and 14% stage III. Group I median survival was 26 months with 30% 5-year survival; for group II, median survival was 30 months and 32%. Kaplan-Meier survival plots showed similar curves in groups I and II. Realizing less extent of disease in group II, another Kaplan-Meier plot restricted to stage I and II patients showed overlapping survival curves for groups I and II. CONCLUSION: Survival during 10-year observation was similar for patients with pulmonary insufficiency treated by wedge resection to that of patients receiving standard resection in this single-institution consecutive cohort. PMID- 17409980 TI - The role of gefitinib treatment for Korean never-smokers with advanced or metastatic adenocarcinoma of the lung: a prospective study. AB - PURPOSE: This prospective trial was conducted to evaluate the role of gefitinib in never-smokers with advanced or metastatic adenocarcinoma of the lung. PATIENTS AND METHODS: The main inclusion criteria were stage IIIB/IV adenocarcinoma of the lung and status as a lifetime never-smoker. Patients received a 250-mg single oral daily dose of gefitinib until disease progression, unacceptable toxicity, or patient's refusal. Tumor response was assessed after every two 4-week cycles according to the World Health Organization response criteria. Additional analyses were performed to identify predictors of response and survival. RESULTS: Between August 2003 and March 2005, 72 Korean patients were enrolled; 55 chemotherapy naive, 17 previously treated; 6 male, 66 female; and ECOG PS 0/1/2, 24/42/4. All patients were assessed for response, toxicity, quality of life, and survival. Overall objective tumor response rate was 55.6% (95% confidence interval [CI], 43.4-67.3%). With a median follow-up of 23 months, the median survival time was 19.7 months (95% CI, 18.5-21.0 months) with a 1-year survival rate of 76.3%. The median duration of response was 6.8 months (95% CI, 4.7-9.0 months). Therapy related improvement of symptoms and quality of life was observed within 2 to 4 weeks after the commencement of therapy in the responders. In a multivariate Cox proportional hazard model, good performance status and no prior history of chemotherapy were the two significant predictors of better survival (p = 0.005 and 0.042). CONCLUSION: Gefitinib showed very promising antitumor activity and survival outcome in Korean never-smokers with adenocarcinoma of the lung. It seems to be a good alternative to standard chemotherapy as a first-line therapy for this subgroup. PMID- 17409981 TI - A phase II study of irinotecan and carboplatin in advanced non-small cell lung cancer with pharmacogenomic analysis: final report. AB - PURPOSE: We conducted a phase II study of carboplatin and irinotecan in patients with advanced non-small cell lung cancer (NSCLC). In addition, we studied the correlation between certain genotypes of enzymes involved in irinotecan metabolism with efficacy and toxicity. PATIENTS AND METHODS: Patients with stage IIIB, IV, or recurrent NSCLC received a combination of irinotecan and carboplatin every 3 weeks at a dose of 200 mg/m2 and area under the curve of 5. Pharmacogenomic analysis was performed on several genes of interest (ABCB1, CYP3A4*1B, ERCC2, GSTP1, UGT1A1*28, and XRCC1). RESULTS: Forty-two patients enrolled between December 2001 and January 2004. Six patients achieved partial responses (14%), and 19 (45%) had stable disease. The median progression-free survival was 6.9 months. The median overall survival was 11.7 months, with 1-year overall survival of 42%. The most common toxicities were hematologic; grade 3 or 4 neutropenia was experienced by 26 patients (62%) during treatment, and 15 patients (36%) experienced grade 3 or 4 thrombocytopenia. The homozygous UGT1A1*28 (7/7) genotype was associated with grade 4 neutropenia in three of four patients (75%), but only eight out of 30 (27%) with 6/6 or 6/7 genotypes experienced grade 4 neutropenia (p = 0.09). None of the 14 patients with the GSTP1 I105V A/A genotype had a partial response, as opposed to five out of 19 (26%) of those with the G/A or G/G genotypes (p = 0.057). CONCLUSION: The combination of carboplatin and irinotecan is an active combination in NSCLC, with response rates comparable with other platinum-containing doublets. Further studies with irinotecan should incorporate prospective pharmacogenomic analysis to identify markers for response and toxicity. PMID- 17409982 TI - Phase II study of carboplatin combined with biweekly docetaxel for advanced non small cell lung cancer. AB - BACKGROUND: The combination of carboplatin and docetaxel has been considered one of the standard treatments for advanced non-small cell lung cancer (NSCLC). To investigate a safer and more convenient schedule for outpatient, we conducted a phase II study to evaluate the efficacy and the safety of carboplatin plus biweekly docetaxel for advanced NSCLC. PATIENTS AND METHODS: Patients with stage IIIB, IV, or postoperative recurrent NSCLC with good performance status were administered docetaxel at a dose of 35 mg/m on days 1 and 15 and carboplatin at an area under the curve (AUC) of 6 on day 1 every 4 weeks for at least three cycles. RESULTS: Fifty patients were treated with median of three cycles (range 1 6). Grade 3/4 toxicities included neutropenia in 18 patients (36%), thrombocytopenia in 4 patients (8%), and anemia in 10 patients (20%). No patient experienced febrile neutropenia. Nonhematological toxicities were also mild to moderate, and there were no treatment-related deaths. The overall response rate was 30%, and the disease control rate was 70%. Among the elderly population, 54% of patients achieved partial response. Median progression-free survival was 4.8 months, and median overall survival was 11.8 months. CONCLUSIONS: Biweekly docetaxel plus carboplatin has a similar efficacy and lower toxicity compared with a standard triweekly regimen of docetaxel plus carboplatin, which is a suitable regimen for outpatients, including elderly patients. PMID- 17409983 TI - DHA-paclitaxel (Taxoprexin) as first-line treatment in patients with stage IIIB or IV non-small cell lung cancer: report of a phase II open-label multicenter trial. AB - INTRODUCTION: This prospective, open-label, non-randomized, multi-institutional phase II study was undertaken to assess the antitumor activity and safety of docosahexaenoic acid-paclitaxel (Taxoprexin) as first-line treatment of patients with advanced non-small cell lung cancer. PATIENTS AND METHODS: Chemotherapy naive patients were eligible if they had measurable stage IIIB or IV non-small cell lung cancer. Forty-four patients received docosahexaenoic acid-paclitaxel by intravenous infusion every 21 days. Two doses were evaluated: 1100 mg/m and 900 mg/m. Patients were monitored for toxicity and tumor response. RESULTS: Patients received between one and seven (median, two) cycles of treatment. Twenty-eight courses were administered in the cohort starting at 1100 mg/m and 109 courses at 900 mg/m. The starting dose was reduced to 900 mg/m because of toxicity in the first 13 patients. Subsequently, the most severe toxicity was neutropenia (grade III/IV in 68% of patients treated with 900 mg/m). Forty patients were eligible for assessment of tumor response. Two partial responses (4.5%) were documented, and a further 16 patients (36.4%) had stable disease based on an intent-to-treat analysis. The median duration of survival for all patients was 243 days (range, 154-359) and the 1-year survival rate was 35%. CONCLUSION: As a single-agent, docosahexaenoic acid-paclitaxel has little activity in patients with advanced non small cell lung cancer, with 18 patients (40.1%) achieving either stable disease or a partial response after treatment. Despite the low objective response rate, treatment was associated with survival comparable to that seen with standard platinum-based combination chemotherapy. The dose-limiting toxicity was myelosuppression. PMID- 17409984 TI - Phase II trial of paclitaxel, carboplatin, and topotecan with G-CSF support in previously untreated patients with extensive stage small cell lung cancer: Southwest Oncology Group 9914. AB - PURPOSE: This phase II study (S9914) evaluated the efficacy and toxicity of the three-drug combination of paclitaxel, carboplatin, and topotecan with granulocyte colony-stimulating factor support in previously untreated patients with extensive stage small cell lung cancer. PATIENTS AND METHODS: Patients with newly diagnosed extensive stage small cell lung cancer received topotecan 1.0 mg/m intravenously on days 1 through 4; paclitaxel 175 mg/m intravenously on day 4, and carboplatin AUC = 5 intravenously on day 4, treatments were repeated every 21 days for a maximum of six cycles. All patients also received granulocyte colony-stimulating factor 5 mug/kg/day beginning on day 5 of each cycle. RESULTS: A total of 88 patients were enrolled on the study; 79 patients were assessable for survival and toxicity and 74 patients for response. Objective response was observed in 50 patients (68%; 95% confidence interval [CI]: 56%-78%) with nine patients (12%) achieving a complete response. Median progression-free survival was 7 months (95% CI: 6-8 months) and median overall survival was 12 months (95% CI: 11-14 months). The 1- and 2-year survival rates were 48% (95% CI: 37%-59%) and 20% (95% CI: 11% 29%), respectively. The most common toxicities were hematologic. Grade 3 and 4 neutropenia was noted in 17 (22%) and 27 (34%) patients, respectively. Febrile neutropenia developed in only four patients. Two patients (3%) died of treatment related causes. CONCLUSION: The combination of paclitaxel, carboplatin, and topotecan combined with granulocyte colony-stimulating factor support is an active and reasonably well-tolerated regimen for the treatment of extensive stage small cell lung cancer. PMID- 17409985 TI - Bortezomib (PS-341) in relapsed or refractory extensive stage small cell lung cancer: a Southwest Oncology Group phase II trial (S0327). AB - BACKGROUND: In preclinical models, the proteasome inhibitor bortezomib (PS-341) inhibits the growth of small cell lung cancer (SCLC) by inhibiting the antiapoptotic Bcl-2 signaling pathway. We conducted a phase II trial of PS-341 in previously treated patients with platinum-sensitive and -refractory extensive stage SCLC to determine response rate, toxicity, and survival. METHODS: Patients with histologically confirmed SCLC, measurable disease, Zubrod performance status 0-1, and previous treatment with platinum-based therapy were enrolled. They were stratified by platinum-sensitivity status: sensitive (relapse >90 days after platinum) or refractory (progression during or < or =90 days after platinum). PS 341 was administered at 1.3 mg/m intravenously on days 1, 4, 8, and 11 every 21 days. RESULTS: Of 56 eligible patients, 28 were platinum sensitive and 28 refractory. Twenty-nine patients (52%) had received two or more previous chemotherapy regimens. One platinum-refractory patient had a confirmed partial response. A majority of assessable patients (91%) progressed. Median progression free survival and overall survival were 1 month and 3 months, respectively. Ten patients (18%) discontinued treatment due to adverse events or side effects. CONCLUSION: Although PS-341 induced a response in a patient with platinum refractory disease, it has limited single-agent activity in this heavily pretreated cohort. As shown in preclinical models, testing of PS-341 in combination with an apoptotic trigger such as chemotherapy, is a rational clinical approach. A trial of topotecan plus PS-341 has been initiated to test this concept. PMID- 17409986 TI - A phase I dose-escalation study of ZD6474 in Japanese patients with solid, malignant tumors. AB - INTRODUCTION: ZD6474 (vandetanib) is an orally available inhibitor of vascular endothelial growth factor receptor, epidermal growth factor receptor, and RET receptor tyrosine kinase activity. This study assessed the safety and tolerability of escalating doses of ZD6474 in Japanese patients with solid, malignant tumors. METHODS: Adult patients with solid tumors refractory to standard therapy received a once-daily oral dose of ZD6474 (100-400 mg) in 28-day cycles, until disease progression or unacceptable toxicity was observed. RESULTS: Eighteen patients were treated at doses of 100 mg (n = 3), 200 mg (n = 6), 300 mg (n = 6), and 400 mg (n = 3). Dose-limiting toxicities at the completion of cycle 2 were hypertension (n = 3), diarrhea (n = 1), headache (n = 1), toxic skin eruption (n = 1), and alanine aminotransferase increase (n = 1). A dose of 400 mg/day was considered to exceed the maximum tolerated dose (MTD). Toxicities were manageable with dose interruption and/or reduction. Objective tumor response was observed in four of nine patients with non-small cell lung cancer (NSCLC) at doses of either 200 or 300 mg. Terminal half-life was about 90-115 hours. Plasma trough concentrations achieved steady-state conditions after approximately 1 month of daily dosing. CONCLUSIONS: It was concluded that a dose of 400 mg/day was considered to exceed the MTD, and doses for phase II study were thought to be not more than 300 mg/day. The objective response observed in some NSCLC patients is encouraging for further studies in this tumor type. PMID- 17409987 TI - A phase I evaluation of oral CI-1033 in combination with paclitaxel and carboplatin as first-line chemotherapy in patients with advanced non-small cell lung cancer. AB - PURPOSE: In the United States, lung cancer represents the third most common cancer, causing the most cancer-related deaths, with treatment advances minimally affecting 5-year survivals. Erb-B family receptor elevations are found in many non-small cell lung cancer tumors, making this receptor family a drug target with potential for improving survival. DESIGN: Chemotherapy-naive patients with advanced non-small cell lung cancer were enrolled who had at least one elevated tumor-expressed member of the erb-B family receptors. This dose-finding, multicenter, open-label, phase I study combined chemotherapy with paclitaxel and carboplatin, adding the tyrosine kinase inhibitor CI-1033. Patients were evaluated for toxicity, response, survival, and pharmacokinetics. RESULTS: All 39 patients enrolled were assessable for safety and efficacy. Dose-limiting toxicities (diarrhea, rash, asthenia, and hypotension) occurred at the 200- and 150-mg dose levels of CI-1033; the maximum tolerated dose was 100 mg. Most toxicities were mild to moderate. Pharmacokinetics studies showed that paclitaxel levels were unaffected by CI-1033 and that CI-1033 plasma concentrations were consistent with historical controls. Ten patients (25.6%) achieved partial responses and another 11 (28.2%) had stable disease. In the recommended phase II dose cohort (n = 23), six patients (26%) had partial responses and six (26%) had stable disease. Median survival time was 12.4 months; median progression-free survival was 5.1 months. CONCLUSION: Paclitaxel-carboplatin, combined with CI 1033 at 100 mg/day, was safe and well tolerated. Efficacy and survival results were comparable to those of similar studies in advanced non-small cell lung cancer and therefore warrant additional phase II testing. PMID- 17409988 TI - Brain: the common site of relapse in patients with pancoast or superior sulcus tumors. AB - PURPOSE: We conducted a retrospective analysis to determine the occurrence of brain metastasis with superior sulcus tumors. METHODS AND MATERIALS: We reviewed 685 charts of patients treated for upper lobe lung cancer between 1997 and 2003. Twenty-nine out of 685 patients (4%) had a diagnosis of Pancoast or superior sulcus tumor. The histology includes 11 patients with adenocarcinoma, seven with non-small cell lung cancer (NSCLC), six with squamous cell carcinoma, four with large cell carcinoma, and one with anaplastic carcinoma. Regarding stage at presentation: seven patients had stage IIB, two had stage IIIA, 16 had stage IIIB, and four had stage IV. RESULTS: The median follow-up is 14 months (range 6 70 months). The total occurrence of brain metastasis is seven out of 29 patients (24%). Two patients (stage IV) had brain metastasis at the time of presentation and five patients (stage IIB-III) developed brain metastasis at a median time of 10 months after the diagnosis. Stage associated with brain metastasis after diagnosis is two patients for stage IIB, two for stage IIIA, and one for stage IIIB. For the 25 patients with stage IIB to stage III disease, nine (36%) developed distant metastasis after definitive therapy. Out of these nine patients, five (55%) developed brain metastasis. It was the most common site of distant failure. Histology for seven patients with brain metastasis was four of seven with adenocarcinoma, two of seven with squamous cell carcinoma, and one of seven with NSCLC. CONCLUSION: Brain metatasis may be relatively common at diagnosis. The brain is the frequent site of failure for superior sulcus tumors. We recommend careful surveillance for brain metastasis during and after the therapy. We also recommend obtaining brain imaging prior to surgery in patients receiving induction therapy for the primary tumor. PMID- 17409989 TI - Early detection of bronchial stump insufficiency after bilobectomy. PMID- 17409990 TI - Spontaneous expectoration of primary pulmonary synovial sarcoma. PMID- 17409991 TI - Proteomic strategies for the characterization and the early detection of lung cancer. AB - Biochemical techniques focused on changes in the proteome in benign and malignant conditions have been applied to the study of lung cancer. Although relatively little information is currently available about the human lung proteome, the major goals of the analysis of lung cancer are to better understand tumor biology, to define early detection biomarkers and predictors of tumor behavior, and to identify potential new therapeutic targets. In this review, we summarized the last 10 years of research in this area with emphasis on its application to the early detection of lung cancer. Basic analytical tools, such as two dimensional gel electrophoresis, mass spectrometry, and protein arrays, are described and placed in the context of lung cancer research. PMID- 17409992 TI - Trichomegaly of the eyelashes after treatment with erlotinib in non-small cell lung cancer. PMID- 17409993 TI - Second-line or subsequent systemic therapy for recurrent or progressive non-small cell lung cancer: a systematic review and practice guideline. AB - PURPOSE: This clinical practice guideline, based on a systematic review, evaluates second-line or subsequent therapy for patients with recurrent or progressive non-small cell lung cancer. METHODS: Relevant randomized trials and meta-analyses were identified through a systematic search of the literature. External feedback was obtained from practitioners in Ontario, and the guideline was approved by the provincial Lung Cancer Disease Site Group. RESULTS: Twenty four randomized trials met the eligibility criteria. Two phase III trials demonstrated a significant benefit in overall survival and quality of life (QOL) for single-agent docetaxel. A pooled analysis comparing docetaxel administered weekly versus three-weekly found similar survival between the schedules and a non significant reduction in febrile neutropenia for the weekly regimen. One phase III trial found that single-agent pemetrexed provided similar survival and QOL, compared to docetaxel. Another phase III trial demonstrated that oral topotecan was non-inferior to docetaxel for one-year survival rate, although QOL significantly favored docetaxel over topotecan. Docetaxel-based and other combination chemotherapy regimens have not been shown to be superior to single agent docetaxel. One phase III trial revealed a statistically significant survival and QOL benefit for erlotinib over placebo for patients who were not eligible for further chemotherapy. Modest tumor response rates and symptom control have been demonstrated for gefitinib; however, a statistically significant survival benefit has not been established for gefitinib over placebo. CONCLUSION: Second-line or subsequent therapy with single-agent docetaxel, pemetrexed, or erlotinib offers patients a significant survival and QOL advantage. PMID- 17409995 TI - Bronchioloalveolar carcinoma and lung adenocarcinoma: the clinical importance and research relevance of the 2004 World Health Organization pathologic criteria. AB - INTRODUCTION: Advances in the pathology and computed tomography (CT) of lung adenocarcinoma and bronchioloalveolar carcinoma (BAC) have demonstrated important new prognostic features that have led to changes in classification and diagnostic criteria. METHODS: The literature and a set of cases were reviewed by a pathology/CT review panel of pathologists and radiologists who met during a November 2004 International Association for the Study of Lung Cancer/American Society of Clinical Oncology consensus workshop in New York. The group addressed the question of whether sufficient data exist to modify the 2004 World Health Organization (WHO) classification of adenocarcinoma and BAC to define a "minimally invasive" adenocarcinoma with BAC. The problems of diffuse and/or multicentric BAC and adenocarcinoma were evaluated. RESULTS: The clinical concept of BAC needs to be reevaluated with careful attention to the new 2004 WHO criteria because of the major clinical implications. Existing data indicate that patients with solitary, small, peripheral BAC have a 100% 5-year survival rate. The favorable prognostic impact of the restrictive criteria for BAC is already being detected in major epidemiologic data sets such as the Surveillance Epidemiology and End-Results registry. Most lung adenocarcinomas, including those with a BAC component, are invasive and consist of a mixture of histologic patterns. Therefore, they are best classified as adenocarcinoma, mixed subtype. This applies not only to adenocarcinomas with a solitary nodule presentation but also to tumors with a diffuse/multinodular pattern. The percentage of BAC versus invasive components in lung adenocarcinomas seems to be prognostically important. However, at the present time, a consensus definition of "minimally invasive" BAC with a favorable prognosis was not recommended by the panel, so the 1999/2004 WHO criteria for BAC remain unchanged. In small biopsy specimens or cytology specimens, recognition of a BAC component is possible. However, it is not possible to exclude an invasive component. The diagnosis of BAC requires thorough histologic sampling of the tumor. CONCLUSION: Advances in understanding of the pathology and CT features of BAC and adenocarcinoma have led to important changes in diagnostic criteria and classification of BAC and adenocarcinoma. These criteria need to be uniformly applied by pathologists, radiologists, clinicians, and researchers. The 2004 WHO classification of adenocarcinoma is readily applicable to research studies, but attention needs to be placed on the relative proportion of the adenocarcinoma subtypes. Other recently recognized prognostic features such as size of scar, size of invasive component, or pattern of invasion also seem to be important. More work is needed to determine the most important prognostic pathologic features in lung adenocarcinoma. PMID- 17409996 TI - BAC consensus conference, November 4-6, 2004: epidemiology, pathogenesis, and preclinical models. AB - INTRODUCTION: Human bronchioloalveolar carcinoma (BAC) is a disease with an evolving definition. "Pure" BAC, characterized by a bronchioloalveolar growth pattern and no evidence of stromal, vascular, or pleural invasion, represents only 2 to 6% of non-small cell lung cancer (NSCLC) cases, but up to 20% of NSCLC cases may contain elements of BAC. This imprecise definition makes it difficult to perform epidemiologic analyses or to generate accurate animal models. However, because BAC appears to behave clinically differently from adenocarcinoma, a better understanding of this disease entity is imperative. METHODS/RESULTS: At the BAC Consensus Conference in 2004, our committee discussed issues relevant to BAC epidemiology, pathogenesis, and preclinical models. CONCLUSIONS: Elucidation of molecular events involved in BAC tumorigenesis will allow for more precise epidemiologic studies and improved animal models, which will enable development of more effective treatments against the disease. PMID- 17409997 TI - Radiographic imaging of bronchioloalveolar carcinoma: screening, patterns of presentation and response assessment. AB - Bronchioloalveolar carcinoma (BAC) is a previously uncommon subset of adenocarcinoma with unique epidemiology, pathology, radiographic presentation, clinical features, and natural history compared with other non-small cell lung cancer (NSCLC) subtypes. Classically, BAC demonstrates a relatively slow growth pattern and indolent clinical course. However, in a subset of patients, rapid growth and death from bilateral diffuse consolidative disease occurs within months of diagnosis or recurrence. Recent data suggest that the incidence of BAC is increasing, notably in younger nonsmoking women. The initial radiographic presentation of BAC varies considerably, from single ground glass opacities (GGOs) or nodules of mixed ground glass and solid attenuation to diffuse consolidative or bilateral multinodular disease. The rising incidence of BAC is also reflected in recent lung cancer screening studies employing helical computed tomography (CT), where the differential diagnosis of GGOs includes not only BAC and overt adenocarcinoma, but inflammatory disease, focal fibrosis, and atypical adenomatous hyperplasia. Because advanced-stage BAC presents as measurable mass lesions in fewer than 50% of cases, determination of radiographic response to therapy by standard criteria is often difficult. Here, we review current data regarding the radiographic imaging of BAC: its radiographic presentations in asymptomatic early-stage and in advanced-stage disease, the functional imaging characteristics of BAC, and challenges of response assessment, including evolving opportunities for computer-assisted image analysis. PMID- 17409998 TI - Surgery for bronchioloalveolar carcinoma and "very early" adenocarcinoma: an evolving standard of care? AB - Lobectomy and mediastinal lymph node dissection is the standard surgical management of early stage non-small cell lung cancer (NSCLC) because more limited resections have been associated with a higher risk of local recurrence. Nevertheless, recent lung cancer screening studies have led to the detection of an increasing number of "very early" NSCLC (defined as less than 2 cm in size) and of good-prognosis histologic subtypes, bronchioloalveolar carcinoma (BAC), and adenocarcinoma (AC), mixed subtypes that are potentially appropriate for sublobar resection. The precise indications for sublobar resection remain unclear and are the subject of ongoing clinical trials, but it seems that very early, peripherally located, node-negative AC of a predominantly BAC pattern may be adequately treated in this manner. Multifocal AC and BAC, either synchronous or metachronous, are also effectively treated by complete resection, using limited resections whenever possible. The pneumonic form of BAC, the rarest variant of this disease spectrum, continues to have a poor prognosis despite complete resection. Very limited experience suggests that lung transplantation leads to prolonged survival in highly selected patients with this histologic subtype. To improve our management of very early AC, much more information is needed about the molecular abnormalities of AC and their relationship to clinical outcomes. PMID- 17409999 TI - Systemic therapy of bronchioloalveolar carcinoma: results of the first IASLC/ASCO consensus conference on bronchioloalveolar carcinoma. AB - INTRODUCTION: Bronchioloalveolar carcinoma (BAC) is a subtype of adenocarcinoma of the lung with unique pathological, clinical, and molecular characteristics. METHODS: This consensus conference group reviewed studies performed specifically in BAC and data from patients with BAC who were included in clinical trials of all non-small-cell lung cancer (NSCLC) subtypes. RESULTS: Although BAC as defined by the World Health Organization represents less than 5% of adenocarcinomas, as many as 20% of adenocarcinomas have BAC features. These latter tumors are more likely to have mutations in the epidermal growth factor receptor (EGFR) gene and to be sensitive to the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Although most patients are men and have a history of smoking cigarettes, proportionally more are women and never smokers. Patients with BAC are routinely treated with drugs and regimens appropriate for patients with all subtypes of adenocarcinoma of the lung; four studies have been performed specifically in this disease. CONCLUSIONS: There is insufficient evidence to confirm or refute the assertion that the sensitivity of BAC to chemotherapy is different from that of other lung cancer histologic types. The unique clinical and molecular characteristics associated with BAC led this panel to conclude that future clinical trials should be designed specifically for persons with BAC. Recommendations for trial design and research questions are proposed. PMID- 17410000 TI - Molecular biology, genomics, and proteomics in bronchioloalveolar carcinoma. AB - The charge of the Molecular Biology, Genomics, and Proteomics in Bronchioloalveolar Carcinoma Committee was to evaluate the molecular biology, genomic changes, and proteomic findings in patients with bronchioloalveolar carcinoma compared with other types of lung cancer. The literature was reviewed and unpublished information was presented by the committee members at the session. The molecular biology studies have included findings on epidermal growth factor receptor (EGFR) mutations, p53 mutations, K-ras mutations, and loss of heterozygosity. The genomic changes have mostly focused on the mRNA expression arrays as well as protein studies. The current state of knowledge was reviewed, the missing information was acknowledged, and proposals for future research were identified. PMID- 17410001 TI - Small randomized trials. PMID- 17410002 TI - Proposals for changes in the Mountain and Dresler mediastinal and pulmonary lymph node map. AB - Staging of the intrathoracic mediastinal and pulmonary lymph nodes is one of the most important problems in the treatment of non-small cell lung cancer. The map of the mediastinal and pulmonary lymph nodes proposed by Clifton F. Mountain and Carolyn M. Dresler gained wide acceptance and has become a standard of staging. However, it has some shortcomings of clarity in the description of localization of specific nodal stations. In our opinion, some modifications of this map are necessary. Based on our experience with extended mediastinoscopy and the new procedure, transcervical extended mediastinal lympadenectomy, the main changes we propose are: 1) the left innominate vein as the anatomic separation between nodal stations 1 and 2; 2) the merging of station 2 and 4 in a single right and left paratracheal station; 3) the shift of the midline to the left paratracheal margin; 4) the tracheobronchial angles as the landmark between stations 4 and 10 bilaterally; 5) the separation of three nodal groups in the subcarinal area: subcarinal (number 7), peribronchial (number 10R and 10L), and periesophageal (number 8); 6) the merging of station 5 and station 6 nodes in a single station with the following landmarks: medial border: the midline, lateral border, the descending aorta and upper border: the left innominate vein and lower border: the lower margin of the left pulmonary artery; and 7) the definition of station 3A nodes as those in front of the superior vena cava. PMID- 17410003 TI - Hedgehog signaling pathway and lung cancer. AB - Signaling pathways responsible for embryogenesis play a critical role in the maintenance of stem cells in adult life and cellular responses to injury. Dysfunction of the developmental signaling pathways during adult homeostasis leads to various events resulting in the development of neoplasia. We review the biology of the hedgehog signaling pathway and its potential role in the development of lung cancer. PMID- 17410004 TI - Genetic heterogeneity in patients with multiple neoplastic lung lesions: a report of three cases. AB - INTRODUCTION: It is important to determine the relation among the various lesions in patients presenting with multiple malignant lung tumors to define the best treatment approach. A better understanding of the molecular alterations present in the different lesions may help in defining this relation. METHODS: We performed a detailed molecular analysis of several tumor specimens obtained from three patients presenting with multiple lung lesions. Tumor specimens were analyzed for epidermal growth factor receptor (EGFR) and k-ras mutations by direct DNA sequencing. In addition, a genome-wide chromosomal copy number analysis was performed on DNA extracted from the various lesions using array based comparative genomic hybridization. RESULTS: In one case, a deletion of 15 base pairs in exon 19 of EGFR was present in all tumor sites analyzed. Furthermore, a similar pattern of chromosomal aberrations was observed among the various lesions, suggesting that they share the same clonal origin. In the other two cases, in contrast, we identified distinct k-ras genotypes among the various lesions from the same patient. These lesions, moreover, showed different chromosomal aberration patterns, indicating that they may have different underlying pathways of tumorigenesis. CONCLUSION: Our results show that EGFR and k-ras mutation analysis, combined with chromosomal copy number profiling, can help in defining the relationship among different tumors in one patient. PMID- 17410005 TI - Prospective validation for prediction of gefitinib sensitivity by epidermal growth factor receptor gene mutation in patients with non-small cell lung cancer. AB - INTRODUCTION: We evaluated the efficacy of gefitinib monotherapy prospectively in patients with advanced or pretreated non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations. METHODS: Patients with NSCLC were examined for EGFR exon 19 deletion mutations by fragment analysis and for EGFR L858R point mutations by the Cycleave polymerase chain reaction technique. EGFR mutation-positive patients with locally advanced, metastatic, or recurrent/refractory NSCLC that was not curable with surgery or thoracic radiotherapy were candidates for gefitinib treatment administered at 250 mg/day until disease progression. RESULTS: Mutations of the EGFR gene were detected in 27 (41%) of 66 patients. Ten had exon 19 deletion, and 17 had L858R. Twenty-one patients harboring EGFR mutations were treated with gefitinib and were considered assessable for responses and adverse events. Nineteen patients with EGFR mutations achieved objective responses (three complete responses and 16 partial responses), resulting in an overall response rate of 90.5% (95% confidence interval, 69.6%-98.8%). The median progression-free survival was 7.7 months (95% confidence interval, 6.0 mo to not reached). The median overall survival has not been reached. Common adverse events were skin toxicity, diarrhea, and elevated aminotransferases, but no pulmonary toxicity was observed. CONCLUSIONS: Detection of common EGFR mutations seems to be useful for selecting patients with NSCLC who would likely benefit from gefitinib monotherapy. PMID- 17410006 TI - The ratio of the maximum high attenuation area dimension to the maximum tumor dimension may be an index of the presence of lymph node metastasis in lung adenocarcinomas 3 cm or smaller on high-resolution computed tomography. AB - BACKGROUND: Limited resection of the lung can improve the postoperative quality of life if there is no lymph node metastasis. Small adenocarcinomas frequently appear on radiographs with areas of ground glass attenuation. We examined whether there is a correlation between the presence of lymph node metastasis and the ratio of the intratumor high-attenuation area (greatest dimension of high attenuation area [H] to the maximum tumor dimension [T], H/T ratio) on high resolution computed tomography (HRCT). METHODS: From January 1996 to June 2004, we studied 281 patients with lung adenocarcinomas of 3 cm or less in maximum dimension on preoperative HRCT. Each of the 281 patients had a lobectomy and lymph node dissection, and there was no case of limited resection. RESULTS: Lymph node metastasis was present in 11.4% (32 of 281 cases, N1 = 13 cases, N2 = 19 cases). None of the 94 cases with an H/T ratio of 60% or less had lymph node metastasis. Moreover, lymph node metastasis was noted even in a 6-mm tumor when the H/T ratio was 100%. Of the 187 cases with tumors having an H/T ratio greater than 60%, 32 had lymph node metastasis. CONCLUSIONS: The relationship between the H/T ratio and lymph node metastases was evaluated by HRCT in lung adenocarcinomas 3 cm or smaller in maximum dimension. The H/T ratio may provide useful information in deciding on limited resection for lung cancer in such cases. PMID- 17410007 TI - Risk factors of postoperative respiratory infections in lung cancer surgery. AB - BACKGROUND: Postoperative infections have been a major issue in lung cancer surgery. We changed our perioperative prophylactic antibiotic policy to a single dose of cefazolin before and after surgery in July 2002. OBJECTIVE: To identify the risk factors of postoperative pneumonia and empyema in lung cancer patients undergoing surgical resection. METHODS: From July 1992 through September 2003, 2105 patients underwent primary lung cancer resection at our division. We reviewed 1855 eligible patients for possible risk factors of pneumonia and empyema. RESULTS: Postoperative respiratory infections developed in 69 (3.7%) patients. There were 58 (3.1%) pneumonia cases and 18 (1.0%) cases of empyema. The mortality rate was 0.8% (15 patients). Nine (0.5%) patients died from postoperative respiratory infections. Multivariate analysis showed age 75 years or older, forced expiratory volume in 1 second as a percentage of forced vital capacity (FEV1%) less than 70%, advanced pathologic stage, and induction therapy to be independent risk factors of pneumonia. For postoperative empyema, advanced age was the significant factor. Twelve of 18 patients (67%) with empyema were complicated with bronchopleural fistula. The infection incidence rate did not change significantly after we modified our prophylactic antibiotic policy to a single dose of cefazolin before and after surgery. CONCLUSIONS: Lung cancer patients with advanced age, low FEV1%, advanced pathologic stage, or induction therapy had a risk for pneumonia after lung cancer surgery. Postoperative empyema was associated with advanced age. PMID- 17410008 TI - Referral patterns for adjuvant chemotherapy in patients with completely resected non-small cell lung cancer. AB - BACKGROUND: Lung cancer remains a leading cause of cancer-related mortality in North America. Despite potentially curative resection, non-small cell lung cancer (NSCLC) patients remain at high risk of relapse and death, with a 5-year survival rate of less than 67%. Several randomized trials now confirm a survival benefit with adjuvant platinum-based chemotherapy seen in the NSCLC Collaborative Group meta-analysis, including the International Adjuvant Lung Trial, National Cancer Institute of Canada BR.10, and Adjuvant Navelbine International Trialist Association (ANITA) trials, with absolute improvements in 4- and 5-year survival rates of 4% to 15%. This study examines whether referral patterns for adjuvant therapy in NSCLC have changed since the presentation of confirmatory trials. METHODS: Retrospective chart review was undertaken at a major tertiary care center, identifying patients with completely resected stages I-IIIA NSCLC from May 2003 to May 2005. RESULTS: A total of 204 patients were identified (59 IA, 77 IB, 8 IIA, 41 IIB, and 19 IIIA). Institutional policy before May of 2003 was not to administer adjuvant therapy outside a clinical trial. After presentation of the International Adjuvant Lung Trial in May 2003, 31% (36/115) of patients with completely resected NSCLC from May 2003 to May 2004 were referred for adjuvant chemotherapy. After presentation of the BR.10 and Cancer and Leukemia Group B 9633 results in June 2004, 63% (56/89) were referred between June 2004 and May 2005. Reasons for not referring to a medical oncologist included stage IA disease, surgeon thought adjuvant therapy inappropriate, patient declined, comorbidities, postoperative complications, and advanced age. Of 92 patients referred to medical oncology, 42 (46%) received adjuvant chemotherapy. Reasons for not prescribing adjuvant chemotherapy included patient refusal (50%), comorbidities (14%), stage IA (10%), and advanced age (4%). Vinorelbine/cisplatin was the regimen most commonly used (67%). CONCLUSIONS: The presentation of positive adjuvant therapy trials in NSCLC has changed clinical practice substantially, doubling the number of patients with completely resected NSCLC referred for adjuvant chemotherapy since May 2004 (31% versus 63%). Although new evidence to support adjuvant chemotherapy in lung cancer is being disseminated to and accepted by physicians, more patient education and decision support may be required to increase uptake of adjuvant therapy in the early stage NSCLC population. PMID- 17410009 TI - Triple combination chemotherapy with cisplatin, docetaxel, and irinotecan for advanced non-small cell lung cancer: a phase I/II trial. AB - BACKGROUND: To determine the recommended dose and evaluate the response rate and toxicity of triplet chemotherapy using cisplatin, docetaxel, and irinotecan for non-small cell lung cancer (NSCLC) patients with stage IIIB or IV. METHODS: A total of 65 patients (33 men and 32 women) with advanced NSCLC, a good performance status, and 65 years of age or younger were included in these phase I/II studies. The median age was 52 years. Most patients had performance status 1 (49/65) and stage IV disease (49/65). Adenocarcinoma was the most common tumor histology (55 patients). Cisplatin and docetaxel were given on day 1 and irinotecan on day 2; the cycles were repeated every 3 weeks. RESULTS: In the phase I study, the maximum tolerated doses of combination cisplatin/docetaxel/irinotecan were, respectively, 80/60/60 (mg/m) and the recommended doses for the phase II study were determined to be 60/60/60 (mg/m), respectively. The dose-limiting toxicities were neutropenia, neutropenic fever, and diarrhea. In the phase II study, 157 cycles of chemotherapy were delivered to 49 patients (median three cycles per patient). The objective response rate was 57.1% (95% confidence interval: 43.1%-71.1%). The median survival time and the actual 2-, 3- and estimated 5-year survival rates were 17 months, 33%, 25%, and 18%, respectively. Grade 3/4 toxicities consisted of neutropenia (92%), neutropenic fever (45%), nausea/vomiting (27%), diarrhea (35%), and hepatic toxicity (2%); there were no cases of treatment-related death. CONCLUSION: This triplet chemotherapy has shown a promising activity against advanced NSCLC according to admission-based treatment with adequate supportive care. The principal toxicity was neutropenic fever, but supportive care should be explored to reduce this incidence. PMID- 17410010 TI - Standard versus dose-intensified chemotherapy with sequential reinfusion of hematopoietic progenitor cells in small cell lung cancer patients with favorable prognosis. AB - PURPOSE: The combination of ifosfamide, carboplatin, and etoposide (ICE) is highly effective in treating small cell lung cancer (SCLC). Myelosuppression resulting in leukopenia and thrombocytopenia is the dose-limiting toxicity. PATIENTS AND METHODS: This phase 3 study assessed 2-year survival improvement with dose intensification of ICE chemotherapy (ICT) in patients with good prognosis SCLC. Patients received up to six cycles of ICT with filgrastim supported sequential reinfusion of peripheral blood progenitor cells every 14 days, or standard ICE (SCT) every 28 days. RESULTS: Eighty-three patients were randomized to ICT (n = 42) or SCT (n = 41). Median survival was significantly improved with ICT (30.3 mo) versus SCT (18.5 mo; p = 0.001); 2-year survival was 55% for ICT and 39% for SCT (p = 0.151). Time to progression (TTP) was significantly improved, with 15 months for ICT versus 11.1 months for SCT (p = 0.0001). Overall response rates were 100 and 88% for ICT and SCT, respectively (p = 0.0258). SCT was associated with significantly less grade 3 and 4 leukopenia at day 8 (p < 0.0001), less thrombocytopenia at day 14 (p < 0.0001), and more favorable platelet nadir (p < 0.0001). The need for platelet and red blood cell transfusions significantly increased in the ICT group (p < 0.0001). Nonhematologic adverse events in both groups were comparable and mostly grade 1 or 2. CONCLUSION: Patients receiving ICT with filgrastim achieved significant increases in median survival and TTP despite an increased need for transfusions. PMID- 17410011 TI - Endobronchial palliation using Nd:YAG laser is associated with improved survival when combined with multimodal adjuvant treatments. AB - BACKGROUND: Endobronchial neodymium:yttrium aluminum garnet (Nd:YAG) laser therapy has been shown to be safe and effective in palliating large airway malignancies. The role of multimodal therapy with stenting, brachytherapy, chemotherapy, and radiotherapy together with Nd:YAG laser therapy is unclear. AIM: This study aims to confirm the safety and effectiveness of Nd:YAG laser therapy in the contemporary setting and to investigate the effectiveness of multimodal therapy compared with laser alone. METHODS: One hundred fifty-three Nd:YAG laser treatments on 110 patients between 1999 and 2004 were reviewed retrospectively. Symptom scores for dyspnoea, hemoptysis, and cough before and after the procedure were compared. Survival and time to reintervention were analyzed using the Kaplan-Meier method. Multimodality treatment was compared with Nd:YAG laser therapy alone to determine differences in survival and time to reintervention. p values less than 0.05 were considered significant. RESULTS: There were no operative mortalities directly caused by laser intervention, although 30-day mortality was 6.5%. Deaths were attributable to progression of advanced neoplastic processes rather than laser intervention, and 6.5% of patients had some postoperative morbidity. After Nd:YAG laser intervention, 76% of patients reported improvement to dyspnoea, 94% for hemoptysis, and 75% for cough. Median survival after Nd:YAG laser treatment was 6.64 months; 21% of patients required repeated laser treatment. Compared with Nd:YAG laser treatment alone, multimodality treatments significantly prolonged median time to reintervention by 1.7 months (p = 0.002) and prolonged median survival by 4.9 months (p < 0.001) in patients with NSCLC. CONCLUSION: Nd:YAG laser intervention is safe and effective for palliation of endobronchial malignancies. In most cases, it only needs to be performed once. Compared with Nd:YAG laser therapy alone, multimodal treatment prolonged survival. PMID- 17410012 TI - Catheter drainage followed by the instillation of bleomycin to manage malignant pericardial effusion in non-small cell lung cancer: a multi-institutional phase II trial. AB - INTRODUCTION: Malignant pericardial effusion (MPE) causes cardiac tamponade and an extremely poor outcome unless it is well controlled. The effect of pericardial drainage and the intra-pericardial instillation of bleomycin on the control of MPE was examined in this prospective multi-institutional phase II trial. METHODS: In eligible patients with cytologically defined MPE resulting from non-small cell lung cancer, the pericardial effusion of such cases was continuously drained. After complete drainage, 10 mg of bleomycin was then locally instilled via a catheter. The catheter was then removed if the total amount of drainage was less than 30 ml/day. If the catheter could not be removed within 5 days after treatment, bleomycin was administered one more time. RESULTS: Among the 22 patients who entered this trial, the tube drainage catheter was successfully removed from 17 patients with one instillation, whereas four required such instillation twice and one required such instillation three times. No severe adverse events were observed, except for constrictive pericarditis, which resulted in the treatment-related death of one patient. The control rate of pericardial effusion was 95% +/- 0.09 (95% confidence interval). No restoration of pericardial effusion was observed during the follow-up period or until death from any cause. Median survival time from the beginning of the protocol was 17.9 weeks. CONCLUSIONS: Pericardial drainage followed by the instillation of bleomycin was found to be a safe and effective method for the management of MPE associated with non-small cell lung cancer. PMID- 17410013 TI - Multidisciplinary management of lung cancer: how to test its efficacy? AB - The multidisciplinary management of lung cancer has been universally accepted. In France, the multidisciplinary approach for cancer patients is established by law. However, the efficacy of this approach remains theoretical, given that no evaluation criteria have been made available and no previous reports have been published on the prospective follow-up of these patients. The Groupe d' Oncologie Thoracique Azureen carried out a 1-year prospective study on patients discussed during its multidisciplinary weekly meetings, to analyze the concordance between the proposed and administered treatment, the delay of treatment, and the 1-year actuarial survival. Of the 344 patients discussed during the period considered, the therapeutic decision was chemotherapy in 183 patients, surgery in 93, radiochemotherapy in 42, radiotherapy in 14, and supportive care 12. Therapeutic discordance between the planned and the administered treatment was recorded in 15 cases (4.4%), mainly for patient's refusal (seven cases) or poor performance status (five cases). The median delay of treatment was 20 days, shorter for chemotherapy (16 days), and longer for radiotherapy (27 days). The overall 1-year survival rate was 51.4%: 80.4% for stage I, 50.3% for stage II, 37.5% for stage III, and 27.2% for stage IV. For patients for whom discordance of treatment was recorded, a lower survival rate was recorded, without reaching statistical significance (0.07). In conclusion, the efficacy of the Groupe d' Oncologie Thoracique Azureen multidisciplinary management was confirmed, as we believe that a discordant rate of less than 5% and a delay of treatment of 4 weeks can be considered acceptable. Furthermore, a periodic survival evaluation of the population as a whole could provide additional useful information for multidisciplinary groups. PMID- 17410014 TI - Multidisciplinary treatment for advanced invasive thymoma with cisplatin, doxorubicin, and methylprednisolone. AB - BACKGROUND AND OBJECTIVES: Advanced invasive thymomas are not usually manageable by surgical resection and radiotherapy. We reviewed our experience with a multidisciplinary approach and evaluated chemotherapy in the treatment of invasive thymoma. PATIENTS AND METHODS: Seventeen consecutive patients with invasive thymoma were treated with multimodality therapy consisting of chemotherapy, surgery, and/or radiotherapy. Four patients had stage III disease with superior vena cava invasion, nine had stage IVa disease, and four had stage IVb disease. The chemotherapy regimen consisted of cisplatin, doxorubicin, and methylprednisolone (CAMP). Chemotherapy was administered in a neoadjuvant setting to the 14 patients and in an adjuvant setting to the remaining three patients. Surgical resection was intended in all patients. After those treatments, chemotherapy and/or radiation therapy were performed. RESULTS: All but one of the 14 patients with induction chemotherapy responded to the CAMP therapy, and the response rate was 92.9%. Seven of these patients underwent complete remission after surgical resection and chemoradiotherapy, and the others underwent partial remission. All three patients treated with surgical resection and then chemotherapy with or without radiotherapy also achieved complete remission. Tumor progression after multimodality therapy occurred in 10 patients. After retreatment, eight of these patients were alive at the time of analysis, with a median survival time after recurrence of 30 months. The 5- and 10-year overall survival rates for all patients were both 80.7%. The major side effect of CAMP therapy was acceptable neutropenia. CONCLUSIONS: CAMP therapy was highly effective for invasive thymomas, and the multimodality therapy containing this chemotherapy brought about good disease control in the majority of patients. We believe that this multidisciplinary treatment with CAMP therapy, surgery, and radiotherapy is a justifiable initial treatment for patients with advanced invasive thymoma. Furthermore, appropriate treatments are essential for the long term survival of patients with recurrences after multimodality therapy. PMID- 17410015 TI - Bilateral patellar metastases from non-small cell lung cancer. PMID- 17410016 TI - A rare presentation of a primary mediastinal lymphoma. PMID- 17410017 TI - A critical look at the role of chemotherapy in older patients with non-small cell lung cancer. AB - Lung cancer remains the leading cause of cancer-related death in the United States, and, at the same time, the American population is getting older. These two factors have given rise to a greater focus on how best to treat non-small cell lung cancer in older patients with cancer. Recent studies have defined the benefits of chemotherapy in older patients with metastatic cancer, and the use of single agent therapy is well justified. In this review, we focus on the need to initiate clinical trials that are specifically designed for older patients with cancer, the role ongoing efforts to identify prognostic factors, the importance of studying newer agents up front in older patients, and the value of phase III trials in this patient population. PMID- 17410018 TI - Unusual chemosensitivity of advanced bronchioalveolar carcinoma after gefitinib response and progression: a case report. AB - Bronchioalveolar carcinoma of the lung represents increasingly recognized clinical entity with relatively high probability of response to epidermal growth factor receptor tyrosine kinase inhibitors. Patients who respond to these agents eventually develop resistance. In this case report, we describe a patient who relapsed after gefitinib treatment and achieved unusual response to vinflunine single-agent chemotherapy, despite earlier progression to a combination of another vinca alkaloid and cisplatin. Molecular characterization of the primary tumor before any treatment is provided, and mechanisms of resistance to epidermal growth factor receptor tyrosine kinase inhibitors are briefly discussed. PMID- 17410019 TI - Surgical resection of esthesioneuroblastoma metastasis to the chest wall. AB - A 51-year-old female patient was evaluated for a painful chest wall mass causing atelectasis of the right lung, pleural effusion, and dyspnea. The patient's history was significant for esthesioneuroblastoma at the age of 24; multiple recurrences of the tumor had been treated with surgery, radiotherapy, and chemotherapy. Surgical resection of the chest wall mass relieved her symptoms and improved her quality of life. Histologic examination confirmed metastatic esthesioneuroblastoma. The patient developed generalized disease and finally died 2 years after surgery. This case demonstrates the long natural history of this rare neoplasm and the need for close follow-up of patients so that they can be treated early. PMID- 17410020 TI - Safety profile of erlotinib in patients with advanced non-small cell lung cancer with chronic renal failure. PMID- 17410021 TI - Long-standing debate on cisplatin- versus carboplatin-based chemotherapy in the treatment of advanced non-small cell lung cancer. PMID- 17410022 TI - Wanted--and Needed--more studies in geriatric lung cancer patients. PMID- 17410023 TI - Immunoinflammatory mechanisms in lung cancer development: is leptin a mediator? AB - This is a short review focusing on leptin immunoinflammatory mechanisms that ultimately may contribute to lung cancer development. We explored the complex and intricate interaction of leptin with immune cells to propose a pathway of inflammation-associated lung cancer development. PMID- 17410024 TI - Nonsteroidal anti-inflammatory drugs and lung cancer risk: a population-based case control study. AB - BACKGROUND: It remains unknown whether nonsteroidal anti-inflammatory drugs (NSAIDs) confer a protective effect against lung cancer development. We examined the relationship between NSAID exposure and subsequent lung cancer development in a large statewide, population-based cohort. DESIGN: Nested case-control study in the Tennessee Medicaid population from 1990 to 2000. METHODS: Lung cancer cases were identified using a statewide computerized claims database. Each time a case was identified, age- and sex-matched controls were randomly selected from the available source population. A detailed pharmacy database quantified nonsteroidal anti-inflammatory drug (NSAID) exposure during the 5 years before cancer diagnosis, and conditional logistic regression was used to examine the relationship between NSAID exposure and lung cancer development. Identical analyses were repeated in a subpopulation of high-risk individuals with chronic obstructive pulmonary disease (COPD). RESULTS: 303,399 persons were enrolled in the study population. During 1.9 million person-years of follow-up (median 6.3 years per person), 3,370 lung cancer cases were identified. Among those ever using NSAIDs, the adjusted odds ratio of developing lung cancer was 1.03 (95% confidence interval: 0.94-1.12). Among those using >24 months of NSAIDs in the 5 years before lung cancer diagnosis, the adjusted odds ratio for lung cancer was 0.96 (95% confidence interval: 0.82-1.11), and no protective effect was demonstrated in any NSAID exposure stratum. Similarly, among the 2519 high-risk individuals with chronic obstructive pulmonary disease, no significant protective effect from NSAIDs was noted. CONCLUSION: This large statewide, population-based study did not demonstrate a protective effect of NSAIDs on lung cancer development in either the general or high-risk chronic obstructive pulmonary disease populations. PMID- 17410025 TI - Primary lung cancer presenting with gastrointestinal tract involvement: clinicopathologic and immunohistochemical features in a series of 18 consecutive cases. AB - BACKGROUND: Lung cancer initially manifesting as gastrointestinal (GI)-tract metastasis is exceedingly rare, representing a diagnostic challenge and a late stage disease sign. The clinicopathologic characteristics of the largest series of lung carcinomas initially presenting with GI involvement were described, focusing on differential diagnosis and therapeutic options. METHODS: Eighteen consecutive cases of lung cancer (11 surgical specimens and 7 biopsies) initially diagnosed on GI histologic samples were identified during routine pathologist practice. All cases were immunostained with thyroid transcription factor-1 (TTF 1), caudal-related homeobox 2 (CDX2), and cytokeratins 7 (CK7) and 20 (CK20). Clinical and radiological data were obtained in all cases. RESULTS: There were 10 women and 8 men with a mean age of 68.5 years. The small bowel was the most common GI involved site (12 cases), followed by the stomach (four) and large intestine (two). Only half of cases were correctly diagnosed on GI biopsies. Fourteen patients died shortly from disease (mean follow-up, 3 months); two are still alive with multiple metastases, and two patients with the GI tract as the unique site of metastasis underwent pulmonary lobectomy and chemotherapy and are alive without evidence of disease. At morphology, there were 10 large cell undifferentiated carcinomas and eight adenocarcinomas. All cases were immunostained for CK7 and 89% for TTF-1, whereas CK20 and CDX2 were completely negative. CONCLUSION: Lung cancer presenting as GI-tract metastasis is probably more frequent than expected, and pathologists should always keep in mind this possibility when dealing with undifferentiated GI carcinoma. Immunostaining with TTF-1, CDX2, CK7, and CK20 is helpful in highlighting lung primary. Although GI metastasis from lung cancer is associated with dismal outcomes, pulmonary resection coupled with chemotherapy might represent a therapeutic option in selected patients with a solitary GI-tract metastasis. PMID- 17410026 TI - Endobronchial ultrasonography in bronchoscopic occult pulmonary lesions. AB - INTRODUCTION: The diagnostic yield of flexible bronchoscopy for peripheral pulmonary lesions is variable and often limited. Endobronchial ultrasonography (EBUS) has been reported to help localize a bronchoscopic occult pulmonary lesion and thereby improve the diagnostic yield of transbronchial biopsy (TBB). METHODS: We evaluated the yield of EBUS-guided TBB in 50 consecutive patients with a bronchoscopic occult pulmonary lesion. RESULTS: The mean diameter of the lesions was 36.6 mm (SD = 19.7 mm). We could visualize 74% of the bronchoscopic occult lesions with EBUS, and in these patients, a histologic diagnosis on TBB was obtained in 84%. However, the diagnostic yield was very poor for lesions <20 mm. CONCLUSION: EBUS-guided TBB is effective for localizing and diagnosing bronchoscopic occult pulmonary masses > or =20 mm, but its yield remains unsatisfactory for lesions <20 mm. PMID- 17410027 TI - Clinical characteristics and survival of patients with surgically resected, incidentally detected lung cancer. AB - BACKGROUND: There is little information on the clinical characteristics and outcomes of patients with surgically resected, incidentally detected lung cancers. Our hypothesis was that among patients with surgically resected non small cell lung cancer (NSCLC), incidentally detected cancers were common, were less likely to require pneumonectomy, and were associated with better stage adjusted survival. METHODS: Two hundred seventy-four patients with NSCLC who underwent surgical resection between 1999 and 2004 were studied. The clinical characteristics of patients with incidentally detected and symptomatic NSCLC were compared. A proportional hazards model was used to compare the stage-adjusted mortality rate of patients with incidentally detected and symptomatic NSCLC. RESULTS: One hundred patients (36%) had incidentally detected NSCLC. Patients with incidentally detected NSCLC had smaller and earlier-stage cancers, were less likely to undergo pneumonectomy (3% versus 13%, p = 0.005), and were more likely to have bronchioloalveolar carcinoma (15% versus 5%, p = 0.003). Patients with incidentally detected cancers had a stage-adjusted hazards ratio (HR) of mortality of 0.9 compared with symptomatic patients (0.6-1.4, p = 0.64). Patients with cancers detected incidentally on computed tomography (CT) had a stage adjusted HR of 0.5 (0.2-1.5, p = 0.15). CONCLUSIONS: Early-stage NSCLC is commonly detected incidentally. Patients with incidentally detected lung cancers are more likely to have bronchioloalveolar carcinoma histology, less likely to undergo pneumonectomy, and overall have similar stage-adjusted survival compared with symptomatic patients. Patients with cancers detected incidentally by CT scan may have better stage-adjusted survival, but our study was not sufficiently powered to detect this effect. PMID- 17410028 TI - Adjuvant surgery after carboplatin and VP16 in resectable small cell lung cancer. AB - INTRODUCTION: The real benefit of surgical treatment of small cell lung cancer (SCLC) has never been demonstrated, mainly because of the rarity of surgical cases and the difficulty in comparing surgical and medical series for the different classifications systems used by surgeons (tumor, node, metastasis) and medical oncologists and radiotherapists (Veterans Administrations Lung Cancer Study Group). MATERIALS AND METHODS: We prospectively assessed the utility of surgery after chemotherapy (carboplatin plus VP16 with or without ifosfamide) with or without radiotherapy in 23 patients with preoperative diagnosis of resectable stage I to IIIA SCLC. A median of three (range: three to six) courses of chemotherapy were administered. Five pneumonectomies, 12 lobectomies (seven sleeve resections), and two segmentectomies were performed, and all except one received radical lymph node dissection. Four (17%) patients received exploratory thoracotomy. Nine (39%) patients received postoperative thoracic radiotherapy. RESULTS: Pathological stages were complete response in four patients, stage I in seven patients, stage II in seven patients, and stage III in five patients. Thirty-day morbidity and mortality were 9% and 0%, respectively. Surgery-related mortality at 90 days was 9%. Median follow-up was 19 months. Overall and local relapse rates were 52% and 17%, respectively. Median overall and disease-free survival were 24 and 12 months. Patients with complete response or pathological stage I had a significantly better Kaplan-Meier survival and lower incidence of relapse than those with more advanced pathological stage (p = 0.025 and 0.027, respectively, log rank). CONCLUSIONS: Survival after chemotherapy and surgery in the series correlated with pathological but not pretreatment stage. Only patients with pathological stage 0 or I disease seem to benefit from surgical resection. PMID- 17410029 TI - Gemcitabine versus gemcitabine-carboplatin for patients with advanced non-small cell lung cancer and a performance status of 2: a prospective randomized phase II study of the Hellenic Cooperative Oncology Group. AB - BACKGROUND: The purpose of this study was to evaluate gemcitabine-carboplatin (GCb) versus single-agent gemcitabine (G) in patients with advanced non-small cell lung cancer (NSCLC) and a performance status (PS) of 2. The primary endpoint was clinical benefit. PATIENTS AND METHODS: Patients were randomly assigned to either 1250 mg/m of G (arm A) or 1250 mg/m of G plus carboplatin area under the curve of 3 (arm B). Both treatments were given on days 1 and 14 and were repeated every 28 days for up to four cycles. RESULTS: Among the 90 eligible patients (47 in arm A and 43 in arm B), in arm A, two (4%) had partial responses (95% CI, 0.52%-14.5%) and 10 (21%) had stable disease (95% CI, 10.7%-35.7%). In arm B, six (14%) had partial responses (95% CI, 5.3%-27.9%) and nine (21%) had stable disease (95% CI, 10%-36%) (p = 0.14). No significant difference was found in terms of clinical benefit between the two treatment groups after two cycles of treatment or at the end of chemotherapy. Furthermore, no association was found between clinical benefit and response to treatment (p > 0.05). Median survival was 4.8 months (95% CI, 2.45-7.25) for arm A and 6.7 months (95% CI, 2.47-10.8) for arm B (p = 0.49). Neutropenia (p = 0.007) and thrombocytopenia (p < 0.001) were more common in group B. Nevertheless, no significant differences were found in terms of severe toxicities (p > 0.05 in all cases). CONCLUSION: No significant difference was found in terms of clinical benefit in patients with NSCLC and PS 2 who received single-agent G or GCb. Nevertheless, GCb caused more toxicity, particularly neutropenia and thrombocytopenia. PMID- 17410030 TI - Many patients 80 years and older with advanced non-small cell lung cancer (NSCLC) can tolerate chemotherapy. AB - People 80 years of age and older constitute 17.8% of all lung cancer patients in the United States. Because the life expectancies of 80-year-old men and women are 87.3 years and 89.0 years, respectively, non-small cell lung cancer shortens lives in addition to causing morbidity. In this retrospective study, all patients with chemotherapy-naive advanced non-small cell lung cancer 80 years of age and older treated at the M. D. Anderson Cancer Center with one or more follow-ups were identified from the database for the years 1997 to 2004. A cohort of patients younger than 80 years old was matched based on treatment year, race, histology, and gender in a 2:1 ratio. Of 13,690 thoracic oncology patients, 496 (3.6%) were 80 years of age and older, of whom 46 met the criteria. In older and younger patients, respectively, platinum doublets were given in 43% versus 79% (p < 0.0001), the response rate was 41% versus 47%, the median progression-free survival was 5.55 versus 3.91 months (p = 0.216), and the median overall survival was 10.7 versus 9.8 months (p = 0.43). Hematologic and nonhematologic toxicities were similar. Our data indicate that selected patients 80 years of age and older may tolerate and benefit from chemotherapy, and prospective evaluation of these patients is indicated. PMID- 17410031 TI - Pemetrexed as second-line treatment in malignant pleural mesothelioma after platinum-based first-line treatment. AB - INTRODUCTION: Pemetrexed is active as first-line treatment of malignant pleural mesothelioma. The objective was to evaluate its activity as second-line treatment. METHODS: Patients had disease progression of malignant pleural mesothelioma after previous platinum-based regimens without pemetrexed. Treatment was pemetrexed alone or pemetrexed combined with carboplatin. Pemetrexed dosing was 500 mg/m and carboplatin was AUC (area under the curve) 5 once every 3 weeks. RESULTS: Thirty-nine patients were included: 28 Danish patients received pemetrexed (three patients received pemetrexed as third-line treatment), whereas 11 Norwegian patients received pemetrexed plus carboplatin. Most patients were men (90%), had epithelial subtype (85%), and International Mesothelioma Interest Group stages III to IV (77%). Median age was 62 years (range, 30-77). The median number of treatment courses was six (range, 1-23). Common Toxicity Criteria grade 3 to 4 toxicity occurred only with respect to leukocytopenia (pemetrexed: 14% of patients; pemetrexed plus carboplatin: 9%) and thrombocytopenia (pemetrexed: 7%; pemetrexed plus carboplatin: 18%). One patient receiving pemetrexed died of sepsis. Partial response rates were 21% and 18%, the median time to progression was 21 weeks (range, 4-92) and 32 weeks (range, 4-128+), and the median survival was 42 weeks (range, 4-99) and 39 weeks (range, 10-128+) with pemetrexed and pemetrexed plus carboplatin, respectively. CONCLUSIONS: Pemetrexed was generally well tolerated with noteworthy activity in malignant pleural mesothelioma after previous platinum-based treatment and may be considered for second-line treatment. PMID- 17410032 TI - A phase II study of paclitaxel, carboplatin, and radiation with or without surgery for esophageal cancer. AB - BACKGROUND: Cisplatin-based chemoradiotherapy (CRT) has been a standard treatment for patients with locally advanced esophageal cancer. However, cisplatin is associated with significant toxicity. We conducted a phase II clinical trial of concurrent paclitaxel, carboplatin, and radiation with or without surgery as an alternative to the standard cisplatin-based CRT for localized and metastatic esophageal cancer. METHODS: Fifty patients with esophageal cancer were enrolled: 16 patients with stage II, eight patients with stage III, and 26 patients with stage IV disease. Two thirds (67%) of patients had adenocarcinoma and one third (33%) with squamous histology. Patients with resectable disease were treated with paclitaxel 30 mg/m, twice weekly for 10 doses, carboplatin AUC (area under the curve) 1.5 weekly for five doses; and concurrent radiation, 1.8 Gy/day, for a total of 45 Gy, followed by esophagectomy. Without surgery, patients received an additional dose each of paclitaxel and carboplatin with concurrent radiation for a total of 50.4 Gy, followed by two consolidation cycles of paclitaxel (200 mg/m) and carboplatin (AUC 6). RESULTS: During CRT, common stage III/IV toxicities included nausea/emesis (19%), esophagitis (9%), and neutropenia (4%). For consolidation chemotherapy, neutropenia (23%), neuropathy (8%) and nausea/emesis (4%) were the most common stage III/IV side effects. After CRT, 26% had a complete response, 17% had a partial response, and 41% had stable disease. Ninety one percent of patients had clinical improvement of dysphagia. With a median follow-up of 32 months, the median survival was 12 months for patients with metastatic disease, 44 months for localized disease treated with esophagectomy, and >44 months for localized disease treated with definitive CRT. CONCLUSIONS: The regimen of paclitaxel, carboplatin, and radiation with or without surgery is well tolerated with promising efficacy for patients with esophageal cancer. PMID- 17410033 TI - Neoplastic fever caused by lung cancer. PMID- 17410034 TI - Amrubicin for the treatment of small cell lung cancer: does effectiveness cross the Pacific? AB - Amrubicin is a synthetic 9-aminoanthracycline that has significant antitumor activity in Japanese patients with extensive stage small cell lung cancer (SCLC). Clinical trials ongoing in the United States and Europe will determine whether amrubicin will be effective in other ethnic groups (whites) or whether this will be an example of geographic and/or genetic variation. Genetic polymorphisms in the UGT1A1 gene have been identified as one of the causes of the increased diarrhea seen in white patients treated with irinotecan when compared with Japanese patients. Nicotinamide adenine dinucleotide phosphate, reduced form quinone oxidoreductase (NQ01) is an enzyme that participates in the metabolism of amrubicin and polymorphisms of the enzyme, known to occur in the Asian population, might explain the effectiveness of the drug in Japanese patients with small cell lung cancer. Studies to evaluate the drug in US and European patients with extensive stage small cell lung cancer are ongoing. Levels of NQ01 will also be determined in these studies. PMID- 17410035 TI - Orbital metastasis as a rare form of clinical presentation of non-small cell lung cancer. AB - Approximately 11 million new cases of cancer are diagnosed worldwide each year; one in eight is a lung cancer. More than one million people die of lung cancer each year, and non-small cell lung cancer accounts for approximately 80% of all cases of lung cancer. The incidence of primary tumors that metastasize to the orbit is approximately 7%. In 19% of the cases, there is no history of cancer when the patient presents with ophthalmic symptoms, and in 10%, the primary site remains obscure despite intensive systemic evaluation. We report a rare form of clinical presentation of non-small cell lung cancer in which symptoms caused by orbital metastases were the first manifestations. PMID- 17410036 TI - Spontaneous complete remission of a non-small cell lung cancer associated with anti-Hu antibody syndrome. AB - Anti-Hu antibodies are directed against lung cancer cell antigens. The anti-tumor effect of anti-Hu antibodies has been suggested by several studies demonstrating that patients presenting with anti-Hu antibodies have a longer survival. In this case report, we suggest that the immunology of HuAb paraneplastic syndrome by itself could induce tumor response. PMID- 17410039 TI - Apoptotic signaling pathways in lung cancer. PMID- 17410040 TI - Ethnic disparities in conditional survival of patients with non-small cell lung cancer. AB - PURPOSE: Conditional survival (CS) is an accurate estimate of survival probability for patients who have already survived at least 1 year after diagnosis. The purpose of this study was to determine whether ethnicity plays a role in 5-year CS rates for patients with non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: Using the Surveillance, Epidemiology, and End Results database, we analyzed 96,480 patients with NSCLC diagnosed between 1988 and 1995. Patients were divided into five ethnic groups: White (non-Hispanic), Hispanic, African American, Asian/Pacific Islander, and Native American/Alaskan. Using the life table method, we computed observed 5-year CS rates for patients who had already survived up to 5 years after diagnosis. Results were analyzed by stage, age, sex, and histology. RESULTS: In general, 5-year CS rates increase for all ethnicities as time from diagnosis increases, but African Americans continued to have lower CS rates compared with other ethnic groups, even up to 5 years from diagnosis. When analyzed by stage, Hispanics with stage IV disease showed the greatest improvement in CS rate, increasing to 73% at 5 years from diagnosis. Among patients older than 70 years, African Americans had the lowest CS at 5 years--only 28%, compared with 40% to 47% for other groups. When analyzed by histology, Hispanics with large cell carcinoma had the worst CS rate (35% at 5 years). CONCLUSION: For patients with NSCLC surviving a period of time after diagnosis, 5-year CS rates vary by ethnicity. CS can provide accurate prognostic information for patients with NSCLC who have already survived several years after diagnosis. PMID- 17410041 TI - The guinea pig syndrome: improving clinical trial participation among thoracic patients. AB - BACKGROUND: The purpose of the study was to examine lung cancer patients' knowledge, attitudes, and behavior regarding clinical trials and to develop an effective intervention for increasing patient knowledge and awareness of clinical trials for lung cancer patients. METHODS: Qualitative semistructured, in-depth interviews were conducted with (1) new patients who had not yet interacted with their physicians, (2) existing patients who had participated in a clinical trial, and (3) existing patients who had not been offered a trial. Findings from the interviews led to the creation of a letter that was sent to all new patients before their first appointment, explaining that offering a clinical trial at this hospital was the norm, to expect this discussion, and the option of standard treatment was always available. RESULTS: Results showed new patients and established patients had similar perceptions of trials--the fear of being a guinea pig, offering a trial meant no hope, and misconception of purpose. Existing patients who had participated in a trial expressed positive benefits of trial participation, even if their health did not improve. A year after the letter process was initiated, accrual rates increased 18% and approximately 81% of all eligible lung cancer patients were in a trial. CONCLUSIONS: The letter, addressing patient's preexisting fears about being offered a clinical trial, appears to show some initial success in improving clinical trial accrual. PMID- 17410042 TI - A pharmacogenomic study of docetaxel and gemcitabine for the initial treatment of advanced non-small cell lung cancer. AB - BACKGROUND: Pharmacogenomic profiling is an attractive strategy for individualizing chemotherapy. Several genetic polymorphisms predict the survival of patients with non-small cell lung cancer treated with platinum-based chemotherapy. This phase II clinical trial was performed using a non-platinum based chemotherapy doublet. The impact of previously identified polymorphisms on clinical outcomes was assessed. METHODS: Patients with advanced non-small cell lung cancer who had not received previous chemotherapy were treated with docetaxel 40 mg/m2 on days 1 and 8 and gemcitabine 800 mg/m2 days 1 and 8 every 21 days until disease progression or unacceptable toxicity. A pretreatment blood sample was obtained, and genomic DNA was analyzed for polymorphisms in DNA repair and metabolic genes. RESULTS: Forty-nine patients were enrolled and evaluated for response and survival. The overall radiographic response rate was 38%, and the median survival was 8.6 months. Nonhematologic toxicity was generally mild. Two treatment related deaths occurred: one due to neutropenic sepsis during the first cycle and one due to pulmonary edema after 12 cycles of treatment. Polymorphisms in XPD, XRCC1, and XRCC3 did not significantly predict survival, but trends similar to those reported for platinum-based chemotherapy were observed. The wild type XPD genotype was associated with prolonged survival and a significantly higher risk of grade 4 neutropenia (p = 0.02). CONCLUSION: This regimen of docetaxel and gemcitabine is well tolerated and active for the treatment of advanced non-small cell lung cancer. The impact of XPD polymorphisms on hematologic toxicity is similar to what has been reported for platinum-based chemotherapy. PMID- 17410043 TI - Phase I study of radical thoracic radiation, weekly irinotecan, and cisplatin in locally advanced non-small cell lung carcinoma. AB - BACKGROUND: Irinotecan and cisplatin individually are active in non-small cell lung carcinoma (NSCLC). Each is synergistic with radiation. Dosages of 65 mg/m2 of irinotecan and 30 mg/m2 of cisplatin Q weekly times four every 6 weeks yielded a 36% response rate and median survival of 11.6 months in advanced NSCLC (Jagasia et al.; Clinical Cancer Research 7: 68, 2001). A weekly schedule for each agent (versus less frequent doses) limits toxicity and increases the opportunity for radiosensitization. MATERIALS AND METHODS: We initiated a phase I study of weekly irinotecan and cisplatin during radical thoracic radiation (TRT). Cisplatin was fixed at 25 mg/m2 Q weekly times seven. Irinotecan was dosed initially at 30 mg/m2 per week for 7 weeks and was increased by 10 mg/m2 per week in three- to six-patient cohorts. TRT was administered in 34 single daily fractions to 63 Gy. Eligibility stipulated locally advanced NSCLC; Eastern Cooperative Oncology Group performance status 0 to 1; < or = 10% unintended weight loss; and adequate physiologic indices. RESULTS: Fifteen patients were accrued: nine were stage IIIB, five were stage IIIA, and one had isolated mediastinal node recurrence after prior surgery. Median age was 65 years (range, 47-77). Seven patients received irinotecan at a dose of 30 mg/m2 per week; (dose level 1). Seven other patients received irinotecan at a dose of 40 mg/m2 per week; (dose level 2). The one other patient received irinotecan in doses of 50 mg/m2 per week; (dose level 3). Neutropenic fever occurred in one patient each at dose levels 1 and 2. Grade 4 neutropenia occurred in three patients at each dose level. Transient grade 3 diarrhea occurred in one patient at dose level 1. Esophagitis of grade 3 or higher occurred in one patient each at dose levels 2 and 3. There was one late grade 3 pneumonitis at dose level 2. Delivered irinotecan dose intensity for dose level 1 was 27 mg/m2 per week; for dose level 2, it was 31.4 mg/m2 per week. Nine of 13 evaluable patients (69%) responded. At median potential follow-up of 5 years, 14 have progressed, and 11 have died. Projected median survival is 28 months; one patient who was treated for mediastinal node recurrence remains free from progression at 6 years. CONCLUSION: Weekly irinotecan and cisplatin combined with radical TRT (63 Gy) is active and fairly well tolerated in locally advanced NSCLC. In combination with fixed-dose cisplatin (25 mg/m2 per week), the maximum tolerated dose of irinotecan is 30 mg/m2 per week. PMID- 17410044 TI - A randomized trial comparing immediate versus delayed treatment of anemia with once-weekly epoetin alfa in patients with non-small cell lung cancer scheduled to receive first-line chemotherapy. AB - INTRODUCTION: This study evaluated the safety/efficacy of once-weekly (QW) epoetin alfa measured by quality of life (QOL), hemoglobin (Hb), transfusion incidence, tumor response, and survival in patients with chemotherapy-naive, advanced non-small cell lung cancer (NSCLC). METHODS: Stage IIIB/IV NSCLC patients with Hb > or = 11 to < 15 g/dl scheduled for at least 8 weeks of first line chemotherapy were randomized to subcutaneously receive 40,000 U of epoetin alfa QW at chemotherapy initiation (immediate) or no epoetin alfa unless Hb decreased to < or = 10 g/dl (delayed). The primary efficacy variable was change in QOL for immediate versus delayed intervention. Target accrual was 320 patients. RESULTS: The study was terminated early because of slow accrual; of 216 patients enrolled, 211 were evaluable for efficacy. Hb was maintained in the immediate group, but it decreased in the delayed group (12.9 versus 11.6 g/dl final values, respectively). Numerically, fewer immediate patients required transfusions versus delayed patients. Mean QOL scores, modestly declining in both groups from baseline to final measurement, were not significantly different between groups. Tumor response and median overall survival were similar between groups. Epoetin alfa was well tolerated, with a similar thrombovascular event rate between groups. CONCLUSION: Epoetin alfa in subcutaneous doses of 40,000 U QW, given immediately at chemotherapy initiation for advanced NSCLC, was well tolerated, and it effectively maintained Hb, leading to a reduced transfusion incidence versus delayed epoetin alfa. Overall QOL scores were higher than typical in this population, decreasing slightly during treatment in both groups. Overall survival was similar between groups, with no evidence of a negative effect by early epoetin alfa intervention. PMID- 17410045 TI - Single-agent pemetrexed or sequential pemetrexed/gemcitabine as front-line treatment of advanced non-small cell lung cancer in elderly patients or patients ineligible for platinum-based chemotherapy: a multicenter, randomized, phase II trial. AB - INTRODUCTION: This randomized phase II trial evaluated single-agent pemetrexed or sequential pemetrexed/gemcitabine in patients with non-small cell lung cancer (NSCLC) who were elderly (> or = 70 years) or younger than 70 years and ineligible for platinum-based chemotherapy. METHODS: Chemonaive patients with stage IIIB/IV NSCLC and an Eastern Cooperative Oncology Group performance status of 0 to 2 received either 500 mg/m2 of pemetrexed (day 1, every 3 weeks) for eight cycles, or the same dosage of pemetrexed for cycles 1 and 2 and then 1200 mg/m2 of gemcitabine (days 1 and 8, every 3 weeks) for cycles 3 and 4 (repeated once for a total of eight cycles). All patients were given vitamin B12 and folic acid supplementation. RESULTS: From July 2003 to July 2004, 87 patients (44 pemetrexed; 43 pemetrexed/gemcitabine) received treatment. The median time to progression was 4.5 (95% confidence interval: 3.0-9.3) and 4.1 months (95% confidence interval: 1.7-5.8) for the pemetrexed and pemetrexed/gemcitabine arms, respectively, and the median progression-free survival time was 3.3 months for both arms. Tumor response rates for the pemetrexed and pemetrexed/gemcitabine arms were 4.5% and 11.6%, respectively. The median overall survival time was 4.7 months for the pemetrexed arm and 5.4 months for the pemetrexed/gemcitabine arm, with respective 1-year survival rates of 28.5% and 28.1%. Grade 3/4 hematologic toxicity consisted of neutropenia (4.5% pemetrexed; 2.3% pemetrexed/gemcitabine), febrile neutropenia (4.5% pemetrexed; 4.7% pemetrexed/gemcitabine), thrombocytopenia (4.5% pemetrexed; 7.0% pemetrexed/gemcitabine), and anemia (6.8% pemetrexed; 4.7% pemetrexed/gemcitabine). No grade 3/4 nonhematologic toxicities exceeded 4.7% in either arm. CONCLUSIONS: Single-agent pemetrexed and sequential pemetrexed/gemcitabine have shown moderate activity and are well tolerated as first-line treatments for advanced NSCLC in elderly patients or patients unsuitable for platinum-based combination chemotherapy. PMID- 17410046 TI - A randomized phase II trial of single-agent gemcitabine, vinorelbine, or docetaxel in patients with advanced non-small cell lung cancer who have poor performance status and/or are elderly. AB - BACKGROUND: Patients with poor performance status and/or are elderly are frequently considered a compromised group at high risk of chemotherapy-related morbidities and less likely to benefit from treatment. We aimed to evaluate tolerability and efficacy of three single-agent regimens in these patients. PATIENTS AND METHODS: Patients with advanced non-small cell lung cancer who had performance status 2/3 and/or were aged 70 and older were randomly assigned to receive gemcitabine, vinorelbine, or docetaxel. Objective response, toxicities, and quality of life were evaluated. RESULTS: One hundred thirty-five patients were registered, of whom one was ineligible. Of the 134 patients, 43 received gemcitabine, 45 vinorelbine, and 46 docetaxel. The response rate was 16%, 20%, 22% for gemcitabine, vinorelbine, and docetaxel, respectively. The main grade 3/4 toxicities were fatigue (18%) and neutropenia (16%). There was improvement in global health scores, cough, and dyspnea for all treatment groups. The improvement in dyspnea was most marked in patients with performance status 3. CONCLUSION: There was no significant advantage of any of the treatment arms over the rest. There was benefit seen with improvement of quality of life in patients who were able to receive more cycles of chemotherapy. PMID- 17410047 TI - Four-modality therapy in malignant pleural mesothelioma: a phase II study. AB - BACKGROUND: Treatment approaches in malignant pleural mesothelioma (MPM) patients range from mere palliation to aggressive anticancer therapy, and there is currently no consensus on the optimal therapeutic strategy. In 1999, we began a phase II study to investigate four-modality treatment of advanced stage MPM. METHODS: From 1999 to 2004, 49 patients with International Mesothelioma Interest Group stage II-III MPM underwent four-modality treatment with intrapleural preoperative interleukin-2 (18 x 10(6) UI/day for 3 days), pleurectomy/decortication, intrapleural postoperative epidoxorubicin (25 mg/m2 for 3 days), interleukin-2 (18 x 10(6) UI/day for 3 days), adjuvant radiotherapy (30 Gy), systemic chemotherapy (cisplatin 80 mg/m2 day 1, gemcitabine 1250 mg/m2 days 1 and 8 for up to six courses) and long-term subcutaneous interleukin-2 (3 x 10(6) UI/day on 3 days per week). RESULTS: Patients included 41 men and eight women with a median age of 61 years (range, 41-77). All patients were diagnosed with MPM by thoracoscopy before inclusion. There was no postoperative mortality. Postoperative morbidity included bleeding (n = 1) and arrhythmias (n = 3). After a median follow-up of 59 months (range, 14-81), 13 patients are still alive and the median actuarial survival is 26 months (31 and 21 months for stages II and III, respectively). The 2- and 5-year actuarial survival rates were 60.2% and 23.3%, respectively. Baseline Eastern Cooperative Oncology Group performance status significantly influenced survival time (p = 0.02). CONCLUSION: The four modality treatment that we adopted for advanced-stage MPM was feasible, well tolerated by most of the patients, and produced a favorable median survival. This treatment approach warrants further investigation. PMID- 17410048 TI - Development of pulmonary arteriovenous communications within pulmonary metastases after chemotherapy. AB - The development of acquired pulmonary arteriovenous communications within pulmonary metastases from an extrathoracic malignancy is a rare phenomenon, but it has potentially serious clinical consequences for the patient. We describe two such patients, both of whom developed intrametastatic pulmonary arteriovenous communications after chemotherapy of the pulmonary metastases, and one of whom required coil embolization for control of symptoms related to the right-to-left shunt. In both cases, computed tomographic angiography was useful for the detection and pre-therapeutic evaluation of the arteriovenous malformations. PMID- 17410049 TI - A rare cause of chylothorax and lymph edema. AB - Chylothorax is a form of pleural effusion rarely caused by metastasis of solid tumors. Because chylothorax causes complaints by local compression of the lung, as well as weight loss resulting from loss of triglycerides, it needs thorough investigation. We present the case of gastric carcinoma presenting with a chylothorax and unilateral lymph edema. Although rare, the differential diagnosis of chylothorax should include gastric cancer even in the absence of upper abdominal complaints. PMID- 17410050 TI - Treating tobacco dependence: review of the best and latest treatment options. AB - Globally, an estimated 85% of lung cancer in men and 47% of lung cancer in women is attributable to tobacco smoking. Tobacco dependence treatment remains the most cost-effective way to prevent morbidity and mortality from lung cancer. Several effective pharmacotherapies are available to treat tobacco dependence. However, the long-term effectiveness of these treatments has been limited because the majority of smokers who attempt to stop smoking eventually relapse. Approaching the treatment of tobacco use and dependence as a chronic disease and the development of innovative drug therapies offer new hope for the treatment of tobacco-dependent patients. The diagnosis of lung cancer provides a teachable moment to motivate patients to attempt tobacco abstinence on which clinicians should capitalize. We review the currently available pharmacologic approaches to the treatment of tobacco dependence. PMID- 17410051 TI - Granulocyte colony-stimulating factor-producing malignant pleural mesothelioma. PMID- 17410052 TI - A clinical case of Castleman's disease. AB - Angiofollicular lymph node hyperplasia, or Castleman's disease, is a rare disorder involving lymphoid tissue proliferation that was first described by Castleman in 1956. The etiology of Castleman's disease is unclear; many origins have been proposed, such as immunocompromised states, chronic inflammation or infection, and autoimmune processes. The disease has been classified on clinical grounds (unicentric or multicentric) and by histological appearance (hyaline vascular pattern, plasma cell predominance, or mixed lesions). In more than 70% of cases, Castleman's disease presents as a solitary mediastinal or cervical mass with an indolent course. Whereas the unicentric form is usually benign and curative resection is possible, patients with the multicentric form often have systemic symptoms and a clinically more aggressive, malignant course. We report a case of unicentric Castleman's disease and re-emphasize its importance in the differential diagnosis of mediastinal tumors. PMID- 17410054 TI - The effects of lactoferrin on myelopoiesis: can we resolve the controversy? AB - Lactoferrin (LF) is an iron-binding protein contained in the secretory fluids of mammals and secondary granules of neutrophils. LF is a key element in innate immunity and exhibits a wide spectrum of activity against pathogens. Although the effects of LF on the maturation and effector function of immune system cells are well recognized, its regulatory function on myelopoiesis has been a matter of controversy for a long time. The majority of early studies demonstrated the role of LF as a negative feedback regulator of myelopoiesis. There were also reports that revealed no significant involvement of LF in that process. However, a number of experimental data, together with clinical observations, indicate a stimulatory action of LF in myelopoiesis. The aim of this article is to review the available literature data regarding the effects of LF on myelopoiesis in mice and humans. Possible reasons for opposing views on this subject are presented. PMID- 17410055 TI - [Difficulties in using archival paraffin-embedded tissues for RNA expression analysis]. AB - Formalin-fixed and paraffin-embedded tissue sections are used for routine histopathological diagnostics, but they have increasingly become material for molecular studies of genome and gene expression using molecular biological techniques such as PCR and RT-PCR. A major limitation is the significant degree of degradation and chemical modification of the nucleic acids recovered from fixed tissues. The purpose of this review is to provide an overview of studies on the possibility of using routinely prepared paraffin-embedded tissue sections as a source of messenger RNA. Difficulties in recovering high-quality RNA are mainly connected with the influence of formalin on nucleic acids and the effects of other physical and chemical agents on tissue during preservation and fixation. It is necessary to optimize RNA isolation and polymerase chain reaction conditions. Special attention is paid to the rising need to introduce alternative techniques for the fixation of tissue sections that provide for better preservation of both macromolecules and tissue morphology and for conducting histological diagnostics with molecular studies. PMID- 17410056 TI - [Arginine metabolism in bronchial asthma]. AB - Asthma is a chronic inflammatory disorder of the airways in which many cells and cellular elements play a role. Airway inflammation is associated with an enhanced expression of inducible nitric oxide synthase. This increases nitric oxide production and results in higher levels of NO* gas in exhaled air. Measurement of exhaled nitric oxide is a very useful non-invasive method in the diagnosis and treatment monitoring of asthma. However, the role of nitric oxide in asthma, still under intense debate, should not be regarded only as a consequence of its abundance, but rather as an impairment of the mechanisms that regulate its synthesis and activity, including reducing nitric oxide production by neuronal and endothelial synthase. Arginine is a substrate for both nitric oxide synthase and arginase. Arginase expression in the lung is strongly induced by cytokines, in particular IL-4 and IL-13, which are produced at elevated level in asthmatic airways and which activate inflammatory pathways. Arginase modulates nitric oxide synthase activity and provides a precursor for polyamines (putrescine, spermidine, and spermine) and proline, which stimulate cell growth and collagen synthesis, respectively. Therefore, arginase might also be involved in inflammation-induced airway remodeling in chronic asthma. This review presents arginine homeostasis in asthma and focuses not only on inducible nitric oxide synthase, but also on impairment of constitutive nitric oxide synthase activity and the overproduction of arginase downstream products. PMID- 17410057 TI - [The role of complement in physiology and pathology]. AB - The complement system was discovered over one hundred years ago. It is an essential part of the innate immune system. A group of about 40 proteins assists in phagocytosis and stimulates inflammation. The complement system participates in the defense of an organism against different factors, e.g. microorganisms. There are three pathways of complement activation: the classical, lectin, and alternative. Activation of the complement system leads to the formation of a lytic macromolecule known as the membrane attack complex (MAC). The MAC may damage target cells in a process called bacteriolysis. The host organism is protected against the negative impact of autoimmunity by complement factor H (CFH). Recent experimental studies dealing with the regulation of the complement system suggest that this control process can be genetically determined. Mutations in genes encoding CFH (CFH polymorphism), factor B, and C2, can be crucial for a defective or insufficient regulation of the complement system. This paper surveys recent achievements on the structure and mechanisms of the complement system and shortly reviews the correlation between the complement function and pathogenesis of many diseases, including atypical hemolytic uremic syndrome (aHUS), membranoproliferative glomerulonephritis II (MPGN II), and age-related macular degeneration (AMD). PMID- 17410058 TI - What's new in carotid angioplasty and stenting. PMID- 17410059 TI - Initial 200 cases of carotid artery stenting using a reversal-of-flow cerebral protection device. AB - AIM: Because embolic complications can occur during carotid angioplasty and stenting (CAS), a new device, the Parodi Anti-Emboli System (PAES) was developed to protect the brain from embolization. We describe our initial experience with this device. METHODS: Between September 1999 and December 2003, CAS was performed in 200 consecutive patients (146 men; mean age, 70.4 years) with symptomatic (52%) or asymptomatic (48%) severe carotid artery stenosis (>70%). De novo lesions were present in 169 patients, restenosis in 18, and radiation-induced stenosis in 13. Wallstents were inserted in all cases, with selective predilatation, and the PAES was employed during all CAS procedures. Patients were evaluated by a neurologist before and after CAS. Minor strokes, major or fatal strokes, and myocardial infarctions that occurred within 30 days of the procedure were recorded. RESULTS: The overall technical success rate for CAS using the PAES (with the PAES placed in position percutaneously) was 99%. The overall perioperative stroke and death rate was 1.5%. There were four transient neurologic events after CAS, three of which were related to hemodynamic instability and one to postoperative embolization. CONCLUSIONS: Our experience indicates that CAS using the PAES is safe and effective. The protection device may prevent the debris released by angioplasty from entering the cerebral circulation. Additional studies of this device are warranted. PMID- 17410060 TI - Are there predictive risk factors for complications after carotid artery stenting? AB - AIM: The aim of this study was to identify predictive risk factors for complications during and after carotid artery stenting (CAS). METHODS: A multivariate analysis was performed on the databases of 4 European high-volume centers regarding risk factor distribution between patients presenting with or without neurological complications 30 days after CAS. The cumulative 30-day neurological complication rate (death, major stroke, minor stroke and transient ischemic attack) was 2.8% in the total examined cohort of 3 179 consecutive CAS procedures. The following risk factors were taken into consideration for statistical analysis: age, symptomatic, male gender, nicotine abuse, hypertension, hypercholesterolemia, polyvascular disease, diabetes, restenosis after carotid endarterectomy (CEA)/CAS, calcified internal carotid artery. RESULTS: Symptomatic (P=0.02) or hypercholesterolemic (P=0.02) patients are at significantly increased risk for neurological events 30 days after CAS. Asymptomatic women and men without hypercholesterolemia have the lowest risk on any 30-day neurological complications after CAS. CONCLUSIONS: CAS is a safe technique in experienced hands. Preprocedural neurological complaints and hypercholesterolemia can be defined as predisposing factors for 30-day neurological complications after CAS. PMID- 17410061 TI - How to optimize carotid artery stenting. AB - Carotid angioplasty and stenting is now an alternative to surgical endarterectomy to treat carotid occlusive disease and is becoming mainstream in medical practice. However, the information available, the procedure techniques and the technologies are still evolving and several issues are still being discussed. Mandatory training, familiarity with the indications and contraindications, knowledge of the technology and devices are paramount for the success, however, the devices for carotid stenting are still under development. The impact of stent design seems to be greater than previously appreciated. Carotid plaque morphology may be important for the indication of the procedure. The pre, trans, and postprocedure patient management of the patient is essential for reducing morbidity and mortality. Due to the devastating potential complications, compared with other endovascular and minimally invasive procedures, carotid stenting requires a much more strict scrutiny of the operators training and outcomes, since the improvement in the learning curve is accompanied by a comparative reduction in the complication rates. This article presents a review of the information available on how to optimize carotid stenting. PMID- 17410062 TI - Dealing with complications related to carotid artery stenting. AB - This paper will describe the complications that can occur during and after carotid artery angioplasty and stenting. Etiology, prevention and treatment of procedure related complications will be dealt with. PMID- 17410063 TI - What is the current status of angioplasty vs endarterectomy in patients with asymptomatic carotid artery disease? AB - Most national and/or specialty driven guidelines recommend against the use of carotid angioplasty/stenting (CAS) in the management of otherwise low or standard risk patients with asymptomatic carotid disease outwith the protection of randomised trials. Despite this relatively clear advice, however, an increasing number of registries and single-centre series now include an ever increasing proportion of low-risk asymptomatic patients within their study cohort. Can this practice currently be justified in the absence of any quality randomised trial evidence? This paper reviews the primary and secondary analyses from the two major randomised trials in asymptomatic patients, discusses the current status of CAS and endarterectomy and highlights areas of controversy and consensus. It concludes with an appeal for all surgeons and interventionists to participate in one of the four randomised trials currently being performed around the world, so that future practice is driven by evidence rather than intuition. PMID- 17410064 TI - Risk factors and management of arterial emboli of the upper and lower extremities. AB - AIM: Analysis of risk factors for the outcome of arterial embolism of the extremities (EE). METHODS: Between 1999 and 2003, all patients (n=200) with an EE diagnosed in various departments of the hospital were recruited and analysed retrospectively (single center study). Exclusion criteria were isolated digital emboli, iatrogenic emboli and arterial thromboses. For statistical analysis was used the multivariate nominal regression. RESULTS: There were 138 (69%) leg, and 62 (31%) arm, emboli. Preoperative angiography was performed in 88 patients; a total of 119 (59.5%) cases of incomplete ischemia (leg n=69, arm n=50) were seen. The most common cause of the embolism (73%) was atrial fibrillation (AF). One hundred and seventhyt four patients (87%) were treated by primary surgery. The major amputation rate (lower limb) was 4.3%. Additional arterial emboli were seen in 14 (7%). The mortality rate was 13% (upper extremity embolism 4.8%; lower extremity embolism 16.7%; P=0.021). After discharge, 32.2% of the patients received oral anticoagulation, and 37.9% antiplatelet therapy. The statistical analysis identified postoperative cerebral/visceral thromboembolism as independent risk factor for mortality. CONCLUSIONS: The main risk factor for EE is AF. Hospital mortality is determined by comorbidity and cerebral or visceral embolism. For this reason, effective oral anticoagulation is required, but is possible in only one-third of the patients after discharge. PMID- 17410065 TI - Open surgery of infrarenal aortic aneurysms with iliac involvement: repair should be entirely intra-abdominal. AB - AIM: A review of the literature on the surgical treatment of abdominal aortic aneurysms (AAAs) reveals that aortofemoral bypass (AFB) is used frequently in some centers. The latter series are characterized by higher rates of graft related complications than in those in which AFB is used less frequently. The aim of our study was to evaluate the relative frequency and outcome of different types of bypass grafts in the surgical treatment of AAAs with iliac involvement, in our center and in others. METHODS: Between 1994 and 2004, 190 AAA patients with involvement of the iliac axes underwent elective repair in our department. Surgery was performed via median transperitoneal access. RESULTS: The AAAs extended to the common iliac artery (CIA) in 90.5% of patients. The remaining 9.5% extended to the external iliac artery (EIA). Aorto bi-iliac grafts were used in 159 cases, straight tube grafts in 13, aorto EIA grafts in 15, and AFBs in 3. Overall 30-day morbidity and mortality rates were 12.1% and 2.6%, respectively. At follow-up (mean: 5.6 years), one distal limb infection of an AFB and 4 CIA/EIA aneurysmal enlargements occurred and were repaired accordingly. Secondary patency and 5-year cumulative survival rate were 100% and 80%, respectively. CONCLUSIONS: In this series of AAAs extending to the iliac axes, AFB was used selectively (1.6%), even when the AAA extended to the EIA. This allowed us to maintain direct vascularization of the hypogastric arteries and eliminate the risk of complications associated with inguinal access. We feel, therefore, that for the repair of aortoiliac aneurysms, AFB is rarely indicated. PMID- 17410066 TI - Endovascular treatment of a ruptured aneurysm of the inferior thyroid artery. Case report and literature review. AB - Aneurysms of the inferior thyroid artery are rare. The natural course of these aneurysms seems to be unfavourable, why aneurysm exclusion is recommended in the literature. Open surgical repair is complex why endovascular exclusion seems to be an appealing alternative. We present a patient who developed dysphagia and respiratory distress caused by a ruptured aneurysm of the right inferior thyroid artery. Successful coil embolization of the aneurysm is described along with a review of the literature. Despite the very rare data of these aneurysms, all reported cases of endovascular treatment (n=3) showed favourable results, therefore aneurysm embolization seems to be a feasible and safe alternative to open surgery, especially in emergency cases. PMID- 17410067 TI - Axillary artery aneurysm following closure of haemodialysis fistula. A case report. AB - Aneurysm formation after ligation of traumatic arteriovenous fistulae is a well documented event, but this complication has been scarcely reported in vascular access for haemodialysis. We describe a case report of delayed aneurysm formation following closure of a haemodialysis fistula. A 57 year-old man was found to have a left axillary artery aneurysm 10 years after closure of a brachio-cephalic arterio-venous fistula. Though he was initially asymptomatic, 6 years later, he developed a progressive sense of coldness in the left hand and an increase in size of the aneurysm. The patient underwent an uncomplicated reversed saphenous vein bypass to replace the aneurysm. The possible aetiological factors and management options are discussed and pertinent literature reviewed. PMID- 17410068 TI - Physical activity post myocardial revascularization. ''Will surgery improve my mobility? AB - AIM: The aim of this study is to evaluate the influence of coronary artery bypass graft (CABG) on physical activity (PA) one year postoperative and the influence of the preoperative level of PA on the postoperative PA level. METHODS: In 428 patients, mean age 64.1+/-9.2 (30-84), undergoing an isolated CABG in 2003, pre and one year postoperatively PA was recorded. Preoperatively, 4 patients (0.9%) were in NYHA I, 33 (7.7%) in NYHA II, 334 (78%) in NYHA III and 57 patients (13.4%) in NYHA class IV. PA was recorded following the Honolulu Heart project. For analysis purposes PA levels were classified as sedentary, 41 patients (9.5%) or minimally active, 161 patients (37.6%) were included in a single sedentary group; 202 patients (47.1%). Patients classified as moderately active, 127 patients (29.6%), active, 72 patients (16.8%) and very active, 27 patients (6.3%) were included in a single active group, 226 patients (52.9%). RESULTS: One year postoperatively, 394 patients (92.%) indicated they were angina-free. One hundred seventy-seven patients (41.3%) recorded a better PA-level. One hundred seventy one patients (40%) recorded the same and 80 patients (18.7%) had a lower PA level. Significantly more patients, 63.9% of the sedentary group versus 21.2% of the active group recorded a higher PA level (P=0.00). Multivariate analysis identified a preoperative active level and vascular disease as risk factors for decreased postoperative PA (P=0.00, odds ratio: 8.1) and a sedentary level as a risk factor for no increase (P=0.00, odds ratio 6.8) CONCLUSIONS: Sedentary patients are likely to show an improvement, however, a sedentary lifestyle is a predictor of no increase in PA. Patients with an active level are unlikely to benefit from surgery. PMID- 17410069 TI - Determinants of late outcome after minimally invasive direct coronary artery bypass. AB - AIM: Minimally invasive direct coronary artery bypass (MIDCAB) is a reliable method to revascularize the left anterior descending (LAD) coronary artery. However, a more consistent body of knowledge is needed to assess factors influencing long-term outcome. With this study, we retrospectively investigated the long-term determinants of survival and freedom from cardiac morbidity and revascularization in patients who underwent MIDCAB. METHODS: From 1997 to 2005, 109 patients underwent MIDCAB. Seventy-five (68.8%) presented isolated LAD disease and 34 (31.2%) multivessel disease. The first 57 patients (53.2%) in the series underwent early postoperative angiographic reinvestigation. All 109 patients were subsequently followed-up at our outpatient clinic. Follow-up (mean 50.7 months, range 3-93) was completed in 100% of cases. RESULTS: No in-hospital deaths occurred; 2 patients (1.8%) experienced perioperative myocardial infarction. At early postoperative angiographic reinvestigation, the anastomotic perfect patency rate was 54/57 (94.7%); survival was 100% and 95.8% at 1 and 5 years, respectively. Overall freedom from repeated revascularization was 95.3% and 88.3% at 1 and 5 years respectively; freedom from LAD revascularization was 95.3% and 91.6% at 1 and 5 years, respectively; cardiac event-free survival was 95.3% and 80.8% at 1 and 5 years respectively. At multivariable analysis (Cox regression), women were found to have a higher risk of repeated LAD revascularization (hazard ratio [HR] 30.24; P<0.001); female sex and left ventricular dysfunction were the only predictors affecting long-term cardiac outcome (hazard ratio 29.35; P<0.001 and 5.1; P<0.001), respectively. CONCLUSIONS: A key factor in the long-term success of MIDCAB seems to be appropriate patient selection. Special attention should be reserved for female patients, as they appear to have a worse cardiac outcome and a higher probability of repeated revascularization on LAD. MIDCAB may represent a viable option for treating multivessel disease when complete revascularization is unfeasible or a hybrid procedure is envisaged. PMID- 17410070 TI - Surgical management of aortic valve disease in elderly patients with and without coronary artery disease: influence on quality of life. AB - AIM: The feasibility of coronary artery bypass grafting (CABG) concomitant with aortic valve replacement (AVR) is well established. However, its impact on long term patient-perceived quality of life (QoL) in the elderly remains undefined. METHODS: Retrospective analysis was conducted on 866 patients 65 years of age and over who underwent AVR between October 1976 and December 1999 with a Carpentier Edwards porcine bioprosthesis. This cohort was divided between those who underwent isolated AVR (n=438) and those with AVR and concomitant CABG (AVR+CABG; n=428). Mean age was 77.0+/-6.1 years (range, 65 to 91) in the AVR group and 78.2+/-5.5 years (range, 65 to 93) in the AVR+CABG group. QoL was assessed with the Short Form-36 health survey for survivors at follow-up, which was 97% complete. RESULTS: Operative mortality (OM) was 6.2% (27/438) for the AVR group and 8.9% (38/428) for the AVR+CABG group (P=0.130). The occurrence of hospital complications (P=0.162) and postoperative length of stay (P=0.980) was similar for the 2 groups. Actuarial survival at 10 years was 37.1+/-3.4% for AVR and 38.7+/-4% for AVR+CABG patients (P=0.088). On multivariate analyses, CABG was not a predictor of either OM or long-term survival. QoL was similar for the 2 groups on the summary components: physical health (39.4+/-11.4 versus 40.2+/-12.1; P=0.461) and mental health (50.2+/-10.8 versus 51.9+/-10.1; P=0.103). CONCLUSIONS: Despite the presence of severe coronary artery disease, CABG preserved the long-term QoL in elderly patients undergoing AVR. PMID- 17410071 TI - Lung cancer and cardiovascular diseases: occurrence, comorbidity and surgical timing. AB - AIM: The authors discuss tumor histological type, TNM staging and time of first occurrence of lung cancer, as well as related cardiovascular diseases, underlining type of treatment, surgical timing, surgical mortality, post-surgical complications and potentially additional therapies. METHODS: A total of 189 patients underwent surgery for non-small cell lung cancer (NSCLC) (Stage I-II), 35 (17.5%) of which presented with surgically relevant concurrent cardiovascular disease (8 coronary artery disease, 4 carotid stenosis, 8 obstructing artery disease, 15 aortic aneurysm). In most cases, surgical timing provides for cardiovascular disease treatment completion first, followed by lung resection only afterwards. Alternatively, concomitant cardiovascular and lung cancer treatment averts the need for repeated surgery, even though the intraoperative complications rate is higher, as long as patients are hemodynamically stabile. RESULTS: No remarkable surgical mortality was observed; stay in hospital ranged from 8 to 18 days. A 5-year follow-up was carried out in only 95/189 patients; 11/18 (61%) with concurrent cardiovascular disease and 46/77 (59%) with lung cancer alone are still alive. CONCLUSIONS: In 17% of cases, surgical treatment by the simultaneous and the differentiated approach for cancer and cardiovascular disease proved essential; comorbidity as an identifier of the strong correlation between the two diseases had a major impact on prognosis. PMID- 17410072 TI - Inducible HSP70 levels in thoracic wound fluid indicate myocardial damage after open heart surgery. AB - AIM: Heat shock proteins (HSP) act as chaperones and protect cellular homeostasis. They are induced by multiple forms of cell stress. Nothing is known about their rise in the drainage fluid after cardiac and thoracic operations. METHODS: Forty-five patients undergoing open heart surgery or major lung resections were studied. Levels of inducible HSP70 were measured daily in thoracic wound fluid and blood with ELISA tests in addition to multiple conventional blood and wound fluid parameters. Data were analyzed using analysis of covariance and Spearman's rank-correlation coefficient. RESULTS: Inducible HSP70 was detectable in all thoracic fluid samples. Mean HSP70-level after open heart surgery was 64 ng/mL and 40 ng/mL in patients without extracorporeal circulation (P=0.015). Drainage HSP70 levels were correlated with white blood cell counts in wound fluid (P<0.0001) and blood CK-MB levels (P<0.0001). Correlation coefficients between white blood cells in blood and local HSP-levels were explicitly lower (P=0.0023). There was a steady decrease in HSP levels in drainage fluid from day 1 to day 4 (P<0.0001). CONCLUSIONS: Inducible HSP70 is present in postoperative thoracic drainage fluid. Drainage levels are higher in patients after open heart surgery and are correlated with blood CK-MB levels, indicating cardiomyocyte damage. Our data further suggest that extracellular HSP70 is released also by white blood cells. Additional studies are needed to examine the role of extracellular HSP70 in wound healing processes and myocardial damage. PMID- 17410073 TI - Body mass index and outcome after coronary artery bypass surgery. AB - AIM: Morbidity and mortality after surgical interventions are influenced by different preoperative factors. We investigated the impact of body mass index (BMI) on outcome after coronary artery bypass grafting (CABG). METHODS: A total of 4 749 CABG patients were divided into 4 groups: low BMI (or=35 kg/m(2), n=146). The incidence of severe perioperative complications (heart failure, renal failure or perioperative stroke), 30-day mortality, length of stay (LOS) and long-term survival were compared. A multivariate analysis with BMI, age, gender and Cleveland Clinic risk score as independent variables and 30-day mortality as dependent variable was performed. RESULTS: Compared to patients with normal BMI, low BMI patients had higher incidence of severe complications (12.5 vs 7.0%, P=0.039), higher 30-day mortality (6.2 vs 1.7 %, P=0.001) and inferior cumulative long-term survival (P=0.04). Patients with moderately increased BMI had longer LOS (10.8 vs 9.0 days, P=0.003) but no difference in incidence of severe complications or mortality. Patients with severely increased BMI had a higher incidence of severe complications (12.3 vs 7.0%, P=0.015, longer LOS (13.0 vs 9.0 days, P<0.001), but no significant difference in early or long-term mortality. Low but not high BMI was an independent predictor for 30-day mortality. CONCLUSIONS: The results suggest that low BMI is associated with increased morbidity and mortality after CABG. Overweight is associated with more postoperative complications and longer hospitalisation but not with an increased early or long-term mortality. PMID- 17410074 TI - Selective antegrade cardioplegic perfusion complicated by left main stem dissection. AB - Selective antegrade coronary artery perfusion is a commonly used procedure to obtain myocardial preservation during cardiac surgery. This report describes a patient operated for severe aortic valve stenosis and insufficiency, mitral valve and tricuspid insufficiency. Cardioplegia was administered by selective antegrade coronary artery blood perfusion. Antegrade blood cardioplegia was complicated by dissection of the left coronary main stem. The dissection induced a myocardial infaction and the patient finally died due to heart failure. PMID- 17410075 TI - Lower extremity compartment sindrome following coronary artery bypass. AB - Compartment syndrome is a constellation of symptoms and signs associated with abnormally elevated tissue pressure in the skeletal muscle of the extremities. It is manifested in anatomic locations where muscles are enveloped in fasciae. The case of a lower extremity compartment syndrome in a 71-year-old male patient who underwent coronary artery bypass grafting (CABG) and simultaneous aortic valve surgery is reported. Preoperative evaluation revealed severe peripheral vascular disease. The patient underwent triple CABG using the left internal thoracic artery and two vein grafts. The right great saphenous vein was used for these vein grafts. The aortic valve was replaced with a biologic prosthesis. On postoperative day 1, the patient complained of pain and oedema in the right calf. The next day, symptoms worsened, with marked sensory loss, motor weakness and foot drop in the affected limb. Triplex ultrasonography excluded deep vein thrombosis. Compartment syndrome was diagnosed and successfully managed by fasciotomy. This case illustrates that compartment syndrome may, although rarely, be a complication of CABG. PMID- 17410076 TI - Selective use of right retroperitoneal approach for abdominal aortic aneurysm repair according to anatomical characteristics revealed by abdominal computerized tomography. PMID- 17410077 TI - Intra-aortic balloon entrapment in the descending aorta. PMID- 17410078 TI - Use and misuse of life-tables in vascular surgery reporting: insight by serendipity. PMID- 17410079 TI - Iatrogenic femoral arterio-venous fistula and pseudoaneurysm following catheter insertion for hemodialysis. PMID- 17410080 TI - Familial secundum type atrial septal defect with normal karyotypes. PMID- 17410081 TI - Fatal occupational injuries--United States, 2005. AB - Data from the annual Census of Fatal Occupational Injuries (CFOI), collected by the Bureau of Labor Statistics (BLS), provide information on fatal occupational injuries that occur in the United States. CDC's National Institute for Occupational Safety and Health (NIOSH) uses CFOI data to support research and evaluation activities related to the National Occupational Research Agenda (NORA), a partnership between the public and private sectors to encourage workplace safety and health research. Since 1992, when BLS first introduced CFOI, BLS has annually reported data on fatal occupational injuries from all 50 states and the District of Columbia. For 2005, BLS reported a total of 5,702 work related fatal injuries and a rate of 4.0 deaths per 100,000 workers; compared with 1992, this represents an 8% decline in the number of deaths (from 6,217 in 1992) and a 23% decline in the fatality rate (from 5.2 in 1992). This report summarizes the 2005 data, which indicated that the highest percentages of fatal workplace injuries were attributed to highway incidents, followed by falls, being struck by an object, and homicides. Since 1992, the number of deaths resulting from highway incidents, falls, and being struck by an object has increased, and the number of homicides has decreased. To reduce the number of workplace deaths, transportation measures targeting workers (e.g., truck safety and highway work zone safety) should be enhanced by state and local transportation agencies and coordinated with highway-safety measures for the general public. PMID- 17410082 TI - Prevalence of fruit and vegetable consumption and physical activity by race/ethnicity--United States, 2005. AB - Diets high in fruits and vegetables and participation in regular physical activity are associated with a lower risk for several chronic diseases and conditions. The National Cholesterol Education Program and the American Cancer Society both emphasize lifestyle modifications that include diet and physical activity to reduce disease risk. These are also two of the strategies implemented by states participating in CDC's Nutrition and Physical Activity Program to Prevent Obesity and Other Chronic Diseases. To examine the combined prevalence of 1) consumption of fruits and vegetables five or more times per day and 2) regular physical activity among U.S. adults by race/ethnicity, CDC analyzed self-reported data from the 2005 Behavioral Risk Factor Surveillance System (BRFSS). This report describes the results of that analysis, which indicated that the combined prevalence of these two behavioral strategies was higher among men of multiple/other races (16.5%) compared with non-Hispanic white men (12.6%). In addition, only 12.6% of non-Hispanic black women and 14.8% of Hispanic women, compared with 17.4% of non-Hispanic white women, engaged in these two behavioral strategies. These results underscore the need to promote diets high in fruits and vegetables and regular physical activity among all populations in the United States and among racial and ethnic minority communities in particular. PMID- 17410083 TI - Elimination of measles--South Korea, 2001-2006. AB - Before the introduction of measles-containing vaccine (MCV), measles was endemic in South Korea. MCV became available in South Korea in 1965, and measles, mumps, and rubella (MMR) vaccine was added to the country's national immunization program in 1983, with 1 dose administered at age 9-15 months. In 1997, a second dose of MCV (MCV2) was added to the schedule; recommended ages for administration of MCV1 and MCV2 were 12-15 months and 4-6 years, respectively. However, with insufficient policies and programs in place to promote 2-dose coverage, this 1997 recommendation achieved limited coverage with 2 doses of MCV. In a 2000 seroepidemiologic study among children aged 7-9 years who had been eligible for vaccination since 1997, only 39% had received MCV2 (Korea Centers for Disease Control and Prevention [KCDC], unpublished data, 2001). During 2000-2001, South Korea experienced a measles epidemic that affected tens of thousands of children. In response, in 2001, South Korea announced a 5-year National Measles Elimination Plan. This report describes the activities and summarizes the results of that plan, which enabled South Korea to announce in late 2006 that interruption of indigenous measles transmission had been achieved, making South Korea the first country in the World Health Organization's (WHO) Western Pacific Region declare measles eliminated. PMID- 17410084 TI - Compendium of Animal Rabies Prevention and Control, 2007: National Association of State Public Health Veterinarians, Inc. (NASPHV). AB - Rabies is a fatal viral zoonosis and a serious public health problem. The disease is an acute progressive encephalitis caused by a lyssavirus. Multiple viral variants are maintained in wild mammal populations in the United States, but all mammals are believed to be susceptible to the disease. For purposes of this document, use of the term "animal" refers to mammals. The recommendations in this compendium serve as a basis for animal rabies-prevention and -control programs throughout the United States and facilitate standardization of procedures among jurisdictions, thereby contributing to an effective national rabies-control program. This document is reviewed annually and revised as necessary. These recommendations do not supersede state and local laws or requirements. Principles of rabies prevention and control are detailed in Part I; recommendations for parenteral vaccination procedures are presented in Part II, and all animal rabies vaccines licensed by the U.S. Department of Agriculture (USDA) and marketed in the United States are listed in Part III. PMID- 17410085 TI - Some more equal than others. AB - Publishers have varying policies as to when published articles become freely available to the public, but it seems that within any one publishing group the free access policy should be the same for all authors. PMID- 17410086 TI - The expanding world of histone lysine demethylases. PMID- 17410087 TI - How HIV-1 hijacks ALIX. PMID- 17410088 TI - Under DNA stress, gyrase makes the sign of the cross. PMID- 17410089 TI - A marked end. PMID- 17410090 TI - La sets the tone for telomerase assembly. PMID- 17410095 TI - Generation and characterization of sodium-dicarboxylate cotransporter-deficient mice. AB - The sodium-dependent dicarboxylate cotransporter (NaDC1) has a proposed function of reabsorbing various Krebs cycle intermediates in the kidney and the small intestine. Since Krebs cycle intermediates have been suggested to be important for renal cell survival and recovery after hypoxia and reoxygenation, the transporter may play a role in the recovery of the kidney. Additionally, mutations in the transporter homolog in Drosophila led to fly longevity which was thought to be similar to that induced by caloric restriction (CR). To clarify the role of the sodium dicarboxylate cotransporter in vivo we generated cotransporter deficient mice. These knockout mice excreted significantly higher amounts of various Krebs cycle intermediates in their urine; thus confirming the proposed function to reabsorb these metabolic intermediates in the kidney. No other phenotypic change was identified in these mice, however. Transporter deficiency did not affect renal function under normal physiological conditions, nor did it have an effect on renal damage and recovery from ischemic injury. Additionally, the absence of the transporter did not lead to metabolic or physiological changes associated with CR. Our results suggest that although the sodium dicarboxylate cotransporter is involved in regulating levels of various Krebs cycle intermediates in the kidney, impaired uptake of these intermediates does not significantly affect renal function under normal or ischemic stress. PMID- 17410096 TI - The pathological role of Bax in cisplatin nephrotoxicity. AB - Nephrotoxicity induced by cisplatin involves tubular cell necrosis and apoptosis; the latter of which may be initiated by multiple mechanisms including activation of the intrinsic mitochondrial pathway. In cultured tubular epithelial cells, cisplatin can activate the proapoptotic protein Bax resulting in cytochrome c release, caspase activation, and apoptosis. Definitive evidence for the involvement of Bax in cisplatin nephrotoxicity in vivo, however, is lacking. We analyzed Bax regulation during cisplatin nephrotoxicity in wild-type mice and determined the pathological role of Bax using mice in which this gene was knocked out. In wild-type mice, cisplatin induced Bax in renal tubular cells which became active, accumulated in the mitochondria, and was accompanied by acute kidney injury. Compared with the wild-type mice renal function, as measured by blood urea nitrogen and serum creatinine, was partially but significantly preserved in Bax knockout mice. The number of apoptotic cells was decreased as was general tissue damage. Additionally, cisplatin-induced cytochrome c release was attenuated in the Bax-deficient mice. This significant decrease in apoptosis and in cytochrome c release was also mirrored in primary cultures of proximal tubular cells prepared from Bax knockout animals. Collectively, our results provide compelling evidence for a role of Bax and its related apoptotic pathway in cisplatin nephrotoxicity. PMID- 17410097 TI - The effect of sirolimus- or cyclosporine-based immunosuppression effects on T cell subsets in vivo. AB - While sirolimus (SRL) is thought to be a non-nephrotoxic agent, cyclosporine A (CsA) toxicity is a serious problem in kidney transplantation. We compared the effects of the two drugs on T-helper (Th) subsets in kidney transplant patients. We examined 24 first cadaver kidney recipients equally randomized to receive SRL/mycophenolate mofetil (MMF)/methylprednisolone (MP), or cyclosporine with either MMF or MP. The Th1 and Th2 subsets in peripheral blood were separated based on their production of interferon-gamma (INFgamma) or interleukin (IL)-4/IL 5. The lymphocytes were stimulated with phytohemoagglutinin or with allogenic CD3 depeted and irradiated antigen-presenting cells. Furthermore, the conversion potential of Th0 to Th1 was determined by measuring IL-12 and IL-18 levels after lipopolysaccharide challenge. When peripheral blood lymphocytes taken from SRL treated patients were stimulated by phytohemoagglutinin, there were significantly lower INFgamma-producing cells compared with the lymphocytes taken from patients treated with CsA. The number of IL-4/IL-5-producing cells did not differ among the patient groups. Release of IL-12 but not IL-18 from peripheral lymphocytes following treatment with lipopolysaccharide was significantly lower in the SRL treated patients. These results show that compared with CsA, SRL caused a significant decrease in the Th1 lymphocyte subset associated with a significant reduction of IL-12 release. PMID- 17410098 TI - Analgesic nephropathy. PMID- 17410099 TI - Polyomavirus-associated nephropathy risk in kidney transplants: the influence of recipient age and donor gender. AB - Polyomavirus-associated nephropathy (PVAN) is a frequent cause of kidney transplant failure. We determined the risk factors for biopsy-proven PVAN among 1027 recent kidney transplant recipients by univariate and multivariate analyses. The rate of PVAN was determined over an univariate and multivariate analysis over an average of 30 months of follow-up of patients receiving predominantly living donor grafts with antibody induction and sequential surveillance biopsies to detect subclinical graft disease. Seventy-four transplant recipients were diagnosed with PVAN with the finding made on surveillance biopsy in 40 patients. These 40 cases did not differ from the 34 non-surveillance cases with respect to baseline clinical characteristics or initial histological features. Older recipient age and female donor gender were independent risks associated with PVAN. Factors not linked to PVAN risk included the use and type of induction agent, use of tacrolimus vs sirolimus, the number of human lympocyte antigen (HLA) mismatches, or the frequency of acute rejection. We conclude that PVAN preferentially affects older age patients and allografts from female donors but is unrelated to immunological risk, choice of immunosuppression, or rejection history. PMID- 17410100 TI - Recurrence of ANCA-associated vasculitis following renal transplantation in the modern era of immunosupression. AB - Progressive glomerulonephritis and attendant end-stage renal disease (ESRD) result from antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis. The optimum time of kidney transplantation in patients with ESRD due to ANCA associated vasculitis (AAV) and the risk of renal or nonrenal recurrence of vasculitis after transplantation are unknown. To answer some of these questions, we followed 35 transplant recipients with diagnoses of microscopic polyangiitis (20 patients) and Wegener's granulomatosis (15 patients). The median time from diagnosis to transplantation was 25 months with all patients being in clinical remission. Fifteen patients were ANCA-positive at time of the transplant with 13 preemptive transplants. The most common immunosuppressive strategy included antibody induction, corticosteroid, mycophenolate mofetil, and tacrolimus with acute rejection occurring in eight cases. Overall and death-censored graft survivals were 94 and 100%, respectively, 5 years post-transplantation. Nonrenal relapse occurred in three patients with a satisfactory response to treatment. No clear risk factor to relapse emerged and no detrimental effect to renal function was found. We conclude that transplantation should be considered as the treatment of choice for ESRD due to AAV. Potent antirejection regimes are well tolerated in these patients, are associated with a low risk of recurrence and an absence of AAV-related graft dysfunction. PMID- 17410101 TI - Vascular calcification: contribution of parathyroid hormone in renal failure. AB - Hyperphosphatemia is a driving force in the pathogenesis of vascular calcification (VC) and secondary hyperparathyroidism associated with renal failure. To test for the possible contribution of parathyroid hormone (PTH) to cardiovascular calcification, we removed the parathyroid glands from rats but infused synthetic hormone at a supraphysiologic rate. All rats were pair-fed low, normal, or high phosphorus diets and subjected to a sham or 5/6 nephrectomy (remnant kidney). Control rats were given a normal diet and underwent both sham parathyroidectomy and 5/6 nephrectomy. Heart weight/body weight ratios and serum creatinine levels were higher in remnant kidney rats than in the sham-operated rats. Remnant kidney rats on the high phosphorus diet and PTH replacement developed hyperphosphatemia and hypocalcemia along with low bone trabecular volume. Remnant kidney rats on the low phosphorus diet or intact kidney rats on a normal phosphorus diet, each with hormone replacement, developed hypercalcemia. All rats on PTH replacement developed intense aortic medial calcification, and some animals presented coronary calcification. We suggest that high PTH levels induce high bone turnover and medial calcification resembling Momckeberg's sclerosis independent of uremia. This model may be useful in defining mechanisms underlying VC. PMID- 17410102 TI - Mesangial immune complex glomerulonephritis due to complement factor D deficiency. AB - Complement factor D is a serine protease essential for the activation of the alternative pathway and is expressed in the kidney, adipocytes, and macrophages. Factor D is found at relatively high levels in glomeruli suggesting that this component of the complement cascade could influence renal pathophysiology. In this study, we utilize mice with a targeted deletion of the activating complement factor D gene and compare these results to mice with targeted deletion of the inhibitory complement factor H gene. Eight-month-old mice with a deleted factor D gene spontaneously develop albuminuria and have reduced creatinine clearance due to mesangial immune complex glomerulonephritis. These mesangial deposits contain C3 and IgM. In contrast to the mesangial location of the immune deposits in the factor D-deficient mice, age-matched factor H-deficient mice develop immune deposits along the glomerular capillary wall. Our observations suggest that complement factor D or alternative pathway activation is needed to prevent spontaneous accumulation of C3 and IgM deposits within the mesangium. Our studies show that the complement factor D gene knockout mice are a novel model of spontaneous mesangial immune complex glomerulonephritis. PMID- 17410103 TI - The spectrum of podocytopathies: a unifying view of glomerular diseases. AB - Glomerular diseases encompass a broad array of clinicopathologically defined syndromes which together account for 90% of end-stage kidney disease costing $20 billion per annum to treat in the United States alone. Recent insights have defined the central role of the podocyte as both the regulator of glomerular development as well as the determinant of progression to glomerulosclerosis. We can now place all glomerular diseases within this spectrum of podocytopathies with predictable outcomes based on podocyte biology impacted by temporal, genetic, and environmental cues. This simplified construct is particularly useful to rationalize clinical effort toward podocyte preservation and prevention of progression as well as to focus basic research effort on understanding podocyte biology and for clinical research toward development of practical monitoring strategies for podocyte injury, dysfunction, and loss. PMID- 17410104 TI - Urinary acidification assessed by simultaneous furosemide and fludrocortisone treatment: an alternative to ammonium chloride. AB - Distal renal tubular acidosis (RTA) can lead to rickets in children or osteomalacia in adults if undetected. This disorder is normally diagnosed by means of an oral ammonium chloride-loading test; however, the procedure often leads to vomiting and abandonment of the test. In this study, we assess an alternative, more palatable approach to test urinary acidification. This was achieved by the simultaneous oral administration of the diuretic furosemide and the mineralocorticoid fludrocortisone to increase distal tubular sodium delivery, principal cell sodium reabsorption, and alpha-intercalated cell proton secretion. We evaluated 11 control subjects and 10 patients with known distal RTA by giving oral ammonium chloride or furosemide/fludrocortisone in random order on separate days. One control and two patients were unable to complete the study owing to vomiting after NH4Cl; however, there were no adverse effects with the furosemide/fludrocortisone treatment. The urine pH decreased to less than 5.3 in the controls with both tests, whereas none of the patients was able to lower the urine pH below 5.3 with either test. We conclude that the simultaneous administration of furosemide and fludrocortisone provides an easy, effective, and well-tolerated alternative to the standard ammonium chloride urinary acidification test for the diagnosis of distal RTA. PMID- 17410105 TI - Sodium channels in neuropathic pain--friend or foe? PMID- 17410106 TI - What impact will nanotechnology have on neurology? PMID- 17410107 TI - From bench to bedside--experimental rationale for immune-specific therapies in the inflamed peripheral nerve. AB - Guillain-Barre syndrome and chronic inflammatory demyelinating polyradiculoneuropathy are autoimmune-mediated inflammatory diseases of the PNS. In recent years, substantial progress has been made towards understanding the immune mechanisms that underlie these conditions, in large part through the study of experimental models. Here, we review the available animal models that partially mimic human Guillain-Barre syndrome and chronic inflammatory demyelinating polyneuropathy, and discuss the wide range of therapeutic approaches that have been successfully established in these models of inflammatory neuropathies. Transfer of this preclinical knowledge to patients has been far less successful, and inflammatory neuropathies are still associated with significant morbidity and mortality. We will summarize successful therapeutic trials in human autoimmune neuropathies to provide a vantage point for the transfer of experimental treatment strategies to clinical practice in immune mediated diseases of the peripheral nerve. PMID- 17410108 TI - Stenting for carotid artery stenosis. AB - Stenting has increasingly been used for the treatment of carotid artery stenosis, although it is still unknown whether it is as safe and successful as carotid endarterectomy. Several studies have been published, and the preliminary results have been variable, with evidence both in favor of and against this procedure. In the past few months, primary outcome data have been published from two large European randomized multicenter trials (SPACE and EVA-3S). So far, both of these trials have evaluated whether carotid stenting shows noninferiority compared with carotid endarterectomy in symptomatic patients with severe carotid disease over a period of 30 days after intervention. In this Review, we summarize current knowledge on effectiveness of both procedures, and provide an updated meta analysis based on randomized trial data, including SPACE and EVA-3S. This meta analysis shows a lower procedure-related rate of stroke or death in surgically treated patients. The long-term risk/benefit ratio of carotid stenting remains to be determined. Two other randomized multicenter trials-ICSS and CREST-are ongoing. With an intended sample size of up to 7,000 patients, future meta analyses will allow more-accurate treatment recommendations and subgroup analysis. PMID- 17410109 TI - Drug Insight: the use of melatonergic agonists for the treatment of insomnia focus on ramelteon. AB - Melatonin, a chronobiotic that participates in the control of the circadian system, is known for its sleep-promoting effects, which include shortening of sleep latency and lengthening of sleep duration. As a result of its short half life, melatonin does not exhibit undesirable side effects, and its broad applicability for a variety of sleep problems has been the focus of numerous scientific studies. Melatonin has not, however, received regulatory approval from the US FDA as a drug, because it can be sold freely as a food supplement. Consequently, there has been an active search for patentable melatonin receptor ligands in recent years. Ramelteon, an agonist that acts solely on melatonin MT(1) and MT(2) receptors, is of particular interest, and preliminary research indicates that it holds considerable promise for clinical applications. Ramelteon has been shown to induce sleep initiation and maintenance in various animal models and in clinical trials. In chronic insomnia, ramelteon decreases sleep latency and increases total sleep time and sleep efficiency, without causing hangover, addiction or withdrawal effects. Ramelteon is thought to promote sleep by influencing homeostatic sleep signaling mediated by the suprachiasmatic nucleus. Although ramelteon's metabolism and pharmacokinetics differ from those of melatonin, its safety seems to be sufficient for short-term application. Its long-term effects remain to be determined. PMID- 17410110 TI - A case of inherited erythromelalgia. AB - BACKGROUND: A 15-year-old boy presented with recurrent episodes of erythema and burning pain in the distal extremities, which he had experienced since early childhood. The episodes were triggered by heat or exertion. His medical history revealed an extensive six-generation family history of similar symptoms. INVESTIGATIONS: Neurological examination, MRI brain scan, electromyography, skin biopsy, laboratory blood testing, and DNA analysis. DIAGNOSIS: Juvenile onset primary erythromelalgia. MANAGEMENT: Genetic counseling, and symptomatic management of neuropathic pain. PMID- 17410111 TI - Teaching residents to be teachers. PMID- 17410112 TI - Exposure modeling of high-frequency electromagnetic fields. AB - We developed a geospatial model that calculates ambient high-frequency electromagnetic field (HF-EMF) strengths of stationary transmission installations such as mobile phone base stations and broadcast transmitters with high spatial resolution in the order of 1 m. The model considers the location and transmission patterns of the transmitters, the three-dimensional topography, and shielding effects by buildings. The aim of the present study was to assess the suitability of the model for exposure monitoring and for epidemiological research. We modeled time-averaged HF-EMF strengths for an urban area in the city of Basel as well as for a rural area (Bubendorf). To compare modeling with measurements, we selected 20 outdoor measurement sites in Basel and 18 sites in Bubendorf. We calculated Pearson's correlation coefficients between modeling and measurements. Chance corrected agreement was evaluated by weighted Cohen's kappa statistics for three exposure categories. Correlation between measurements and modeling of the total HF-EMF strength was 0.67 (95% confidence interval (CI): 0.33-0.86) in the city of Basel and 0.77 (95% CI: 0.46-0.91) in the rural area. In both regions, kappa coefficients between measurements and modeling were 0.63 and 0.77 for the total HF-EMF strengths and for all mobile phone frequency bands. First evaluation of our geospatial model yielded substantial agreement between modeling and measurements. However, before the model can be applied for future epidemiologic research, additional validation studies focusing on indoor values are needed to improve model validity. PMID- 17410113 TI - Potential exposure and risk of fluoride intakes from tea drinks produced in Taiwan. AB - Tea is the second most commonly consumed drink in the world. Excess fluoride intakes from tea drinks may cause health effects. This work assesses infusible fluoride levels in popular tea sold in Taiwan and evaluates potential exposure factors. Lungjing, pouchong, tienguanyin, oolong, pureh, and black tea specimens were purchased from different counties in Taiwan. Fluoride levels were evaluated in one complete cycle of tea making as well as at different calcium carbonate contents in water, with glass or porcelain teapots, and with/without adding sugar. Oolong tea leaves in each manufacturing step were also analyzed for infusible fluoride. Potential fluoride intakes and risks are estimated based on a national survey. Among six kinds of tea, black tea had the highest fluoride concentrations (8.64+/-2.96 mg/l), whereas pureh (1.97+/-2.70 mg/l) had the lowest levels. Higher percentages of infusible fluoride can be rinsed away from tea leaves curved lengthways compared to those curved end-to-end in the first 2.5 min. The use of glass or porcelain teapots and calcium carbonate content (up to 400 mg/l) in water would not affect infusible fluoride levels, whereas adding sugar increased the infusible fluoride in the first few minutes. In addition, it was found that the critical step during the manufacturing process affecting the percentage of infusible fluoride was ball rolling rather than fermentation. Furthermore, intakes of high amounts (> or =5 l/week) of certain tea may result in excess risks of dental or skeletal fluorosis. Tea lovers could be exposed to excess fluoride and may be at risk of fluorosis. PMID- 17410114 TI - Temporal stability of the conjugated species of bisphenol A, parabens, and other environmental phenols in human urine. AB - Human exposure to environmental phenols can be assessed by measuring the urinary concentrations of these compounds or their metabolites. Total concentrations, which include both free and conjugated (i.e., glucuronide and sulfated) species, are usually reported. Because conjugation may reduce the potential bioactivity of the compounds, measuring separately both the concentrations of free and conjugated species can be of interest. Data on the stability of these conjugated species in urine is critical if the concentrations of free and conjugated species are to be compared. Over a period of 6 months, we investigated the stability of the urinary conjugates of eight environmental phenols (bisphenol A, 2-hydroxy-4 metoxybenzophenone or benzophenone-3, triclosan, 2,5-dichlorophenol, methyl paraben, ethyl paraben, propyl paraben, and butyl paraben) at three storage conditions (room temperature, 4 degrees C, and -70 degrees C). After collection, conjugated species appeared to be stable for at least 7 days when the urine was stored at 4 degrees C, and for at least 180 days at -70 degrees C. By contrast, some of the environmental phenol conjugates commenced to degrade within 24 h after collection when the urine was stored at room temperature although the total concentrations remained relatively constant for at least 30 days. These results suggest that if the concentrations of free and conjugated species will be used for exposure assessment purposes, urine specimens collected for analysis of environmental phenols should be kept at room temperature for the shortest possible time after collection. PMID- 17410115 TI - An evaluation of semi-quantitative test strips for the measurement of nitrate in drinking water in epidemiologic studies. AB - Consumption of nitrate and nitrite is associated with a variety of health outcomes. Commercially available test strips that allow semi-quantitative estimation of these contaminants in drinking water are inexpensive relative to laboratory testing, and are simple to use. To examine the accuracy of a nitrate/nitrite test strip, we recruited Washington State residents to estimate levels of these contaminants in their tap water using these strips, and simultaneously provide a tap water sample for laboratory analysis. Paired results were available from 102 homes. On the basis of laboratory assay, nitrate levels as nitrogen ranged from no nitrate (27%) to 40.5 mg/l (median 0.4 mg/l). Spearman's correlation coefficient between test strip- and laboratory-measured nitrate indicated moderate precision overall (r=0.72). Correlation was similar for homes inside and outside city/town limits, but differed by primary source of water for the purveyor indicated by residents (r=0.72 for groundwater and r=0.34 for surface water). Seven (7%) participants reported difficulty in distinguishing colors (contaminant levels) when using the test strip; and among the samples with nitrate, the laboratory assay indicated higher nitrate levels than the test strip for 81%. Nitrite was not detected by laboratory assay; in comparison, five (5%) subjects reported any nitrite according to the test strip. Nitrate/nitrite test strips may be useful in some epidemiologic studies, but should be used with caution, preferably as a screening tool or when laboratory assays are not feasible. PMID- 17410116 TI - Relationship between indoor, outdoor, and personal fine particle concentrations for individuals with COPD and predictors of indoor-outdoor ratio in Mexico city. AB - Personal exposure and indoor and outdoor exposure to PM(10) and PM(2.5) of 38 individuals with chronic obstructive pulmonary disease (COPD) was characterized from February through November 2000. All participants lived in Mexico City and were selected based on their area of residence southeast (n=15), downtown (n=15), and southwest (n=8). Participants were monitored at home using personal PM(2.5) monitoring devices. Indoor and outdoor levels of PM(10) and PM(2.5) were measured using MiniVol samplers. Concurrent individual exposure measurements, indoor and outdoor levels of PM(2.5), which averaged 38.4 (SD 21.4), 30.6 (SD 15.8), and 30.5 mug/m(3) (SD 19.4), respectively. Indoor PM(2.5) concentrations explained 40% of the variability of personal exposure. In addition, the factors that most affected personal exposure were regular indoor contact with animals, mold, cooking activities, and aerosol use, indicating that internal sources may largely affect individual exposure. PMID- 17410117 TI - Update of US FDA's Total Diet Study food list and diets. AB - The US Food and Drug Administration's (FDA) Total Diet Study (TDS) has been conducted continuously since the early 1960s to measures levels of various pesticide residues, contaminants, and nutrients in foods and to estimate the dietary exposures to these compounds. Both the TDS food list and the consumption amounts used for estimating exposures are based on results of nationwide food consumption surveys, and they are updated periodically to reflect changes in food consumption patterns. The most recent update was completed in 2003 using the same methodology employed in the previous update (1990). The updated food list includes approximately the same number of foods (285) as the previous list (290). Although most (75%) foods are the same in both versions, the new list reflects trends in consumption of foods containing less fat. The updated diets reflect an increase in total food consumption, with most notable increases in consumption of grains and beverages. A case study comparing cadmium exposures calculated from both the 1990 and 2003 versions of the TDS demonstrated the potential impact of changes in both the food list and consumption amounts on TDS exposure estimates. PMID- 17410118 TI - Pharmacokinetics of 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol (CTHC) after intravenous administration of CTHC in healthy human subjects. AB - After cannabis consumption there is only limited knowledge about the pharmacokinetic (PK) and metabolic properties of 11-nor-9-carboxy-Delta(9) tetrahydrocannabinol (CTHC), which is formed by oxidative breakdown from Delta(9) tetrahydrocannabinol (THC). Despite widely-varying concentrations observed in smoking studies, attempts have been made to interpret consumption behavior with special regard to a cumulated or decreasing concentration of CTHC in serum. Ten healthy nonsmoking white male individuals received 5 mg CTHC intravenously over 10 min. Highest serum concentrations of CTHC were observed at the end of the infusion (336.8+/-61.7 microg/l) followed by a quick decline. CTHC concentration could be quantified up to 96 h after administration, with a terminal elimination half-life of 17.6+/-5.5 h. Total clearance was low (91.2+/-24.0 ml/min), with renal clearance having only a minor contribution (0.136+/-0.094 ml/min). This first metabolite-based kinetic approach will allow an advanced understanding of CTHC PKs data obtained in previous studies with THC. PMID- 17410119 TI - Clinical pharmacology information in summaries of product characteristics and package inserts. AB - The aim of the study was to assess the clinical pharmacology information provided in the summaries of product characteristics (SPCs) of European drugs compared with their package inserts. A list of the 150 most prescribed drugs at the local Health Service was assessed. To analyze clinical pharmacology information contained in package inserts and SPCs, Spyker's systematized scoring approach was used (scoring from 0 to 2). Ninety-one SPCs (84.3%) and 108 package inserts were obtained. The mean score for core information provided in the package inserts was 7.94 (confidence interval (CI) 95 %: 7.34-8.53), whereas for the SPCs it was 17.62 (CI 95 %: 16.48-18.75; 35.2% of the maximum). One core information item and two extra credit items were not contained in any of the SPCs. In the European Union the clinical pharmacology information found in SPCs is clearly insufficient. An improved access to and regular review of the SPCs might substantially improve the access and quality of clinical pharmacology and other prescribing information. PMID- 17410120 TI - Comparison of the effects of thioridazine and mesoridazine on the QT interval in healthy adults after single oral doses. AB - We compared the effects of single doses of thioridazine and mesoridazine on the heart rate-corrected QT (QTc) interval in healthy adult volunteers. QTc intervals and plasma concentrations of thioridazine, mesoridazine, and metabolites were measured after single oral doses of thioridazine hydrochloride 50 mg, mesoridazine besylate 50 mg, or placebo in a double-blind, crossover study. Mean maximum increases in the QTc interval following thioridazine (37.3+/-4.1 ms, P=0.023) and mesoridazine (46.6+/-7.4 ms, P=0.021) were similar and significantly greater than following placebo (12.9+/-8.1 ms). The area under the effect-time curve over 8 h following drug administration was similar between the two drugs (129.3+/-22.1 vs 148.3+/-43.0 ms h). In conclusion, thioridazine and mesoridazine are associated with similar effects on the QTc interval. PMID- 17410121 TI - Pharmacokinetics of quetiapine in overdose and the effect of activated charcoal. AB - The aim of the study was to investigate the pharmacokinetics of quetiapine overdose and the effect of charcoal. The data set included 204 concentration-time points from 54 quetiapine overdose events (median dose 2,700 mg (300-24,000 mg)). Charcoal was administered 0.5-6 h after 19 overdoses. A fully Bayesian methodology for population pharmacokinetic analysis was used and data were modelled using WinBUGS. Uncertainty in the dose history was considered in model building by estimating dose amount and dose time within a possible range. Inclusion of informative priors stabilized the model and population parameter values could be estimated well. A one-compartment model with first-order input and first-order elimination described the data. The final model included uncertainty in dose time. The median and interquartile range of the half-life for individual patients was 6.6 h (4.9-8.4 h). Charcoal was estimated to reduce fraction absorbed by 35%. Co-ingested CYP3A4 inhibitors appeared to decrease clearance and CYP3A4 inducers increase clearance. Charcoal administration may be beneficial after quetiapine overdose. PMID- 17410122 TI - Balancing justice and autonomy in clinical research with healthy volunteers. AB - In clinical research, ethics review generally first examines whether study risks are reasonable in light of benefits provided. Through informed consent, then, prospective subjects consider whether the risk/benefit balance and procedures are reasonable for them. Unique ethics issues emerge in clinical research with healthy volunteers. Certain types of studies only recruit healthy volunteers as participants. Phase 1 studies, for example, including first time in human studies of investigational drugs and vaccines, generally are conducted in healthy volunteers. Although such research carries inherent and often unknown risks, healthy subjects provide the most efficient target population in which to conduct such research, as these volunteers generally are free of concurrent diseases or medications that could confound interpretation of toxicity. Other studies enrolling healthy volunteers often are simply looking for the most scientifically sound population for the study of normal human physiology. PMID- 17410123 TI - Presynaptic alpha-2C adrenoceptor-mediated control of noradrenaline release in humans: genotype- or age-dependent? AB - In vitro alpha-2CDel322-325 adrenoceptor (AR) polymorphism exhibits reduced functional responsiveness. We studied whether this is true also in vivo in humans. We assessed in nine young wild-type (WT) alpha-2C AR subjects (aged 23 years), 10 elder WT alpha-2C AR subjects (aged 63 years), and nine alpha-2CDel AR subjects (aged 28 years) clonidine (1 microg/kg intravenous (i.v.) bolus)-evoked plasma noradrenaline (pNA), heart rate (HR), and blood pressure (BP) changes. Clonidine-evoked pNA decreases were comparable in young WT alpha-2C and in alpha 2CDel AR subjects, but significantly lower (P=0.033) in elder subjects. Similarly, clonidine-evoked HR decreases were significantly larger in young WT alpha-2C and in alpha-2CDel AR subjects than in elder subjects, whereas clonidine evoked BP decreases were larger in elder subjects. In conclusion, alpha-2CDel AR appears to play only a minor role in presynaptic regulation of NA release and/or to be not hypofunctional in vivo in humans, but functional responsiveness of presynaptic alpha-2 AR declines with ageing. PMID- 17410124 TI - Pharmacokinetics and pharmacodynamics of the dual FII/FX inhibitor BIBT 986 in endotoxin-induced coagulation. AB - BIBT986 is a dual inhibitor of factors Xa and IIa. The aim of this study was to compare with placebo the effect of three doses of BIBT986 on coagulation, platelet activation, and inflammation. This was a prospective, randomized, double blind, placebo-controlled, parallel-group dose escalation trial in 48 healthy male volunteers. Participants received one of three doses of BIBT986 or placebo intravenously together with a bolus infusion of 2 ng/kg lipopolysaccharide (LPS). BIBT986 dose-dependently changed global coagulation parameters and in vivo markers of thrombin generation and action: BIBT986 doses, which prolonged activated partial thromboplastin time by 100%, completely suppressed the LPS induced increases in prothrombin fragment, thrombin-antithrombin complexes, and D dimer, which were 6.1-, 14.5, and 3.5-fold in the placebo group, respectively. BIBT986 did not influence inflammation, fibrinolysis, or platelet activation. Therefore, BIBT986 is a potent anticoagulant in the human endotoxemia model. PMID- 17410125 TI - A new perceptual illusion reveals mechanisms of sensory decoding. AB - Perceptual illusions are usually thought to arise from the way sensory signals are encoded by the brain, and indeed are often used to infer the mechanisms of sensory encoding. But perceptual illusions might also result from the way the brain decodes sensory information, reflecting the strategies that optimize performance in particular tasks. In a fine discrimination task, the most accurate information comes from neurons tuned away from the discrimination boundary, and observers seem to use signals from these 'displaced' neurons to optimize their performance. We wondered whether using signals from these neurons might also bias perception. In a fine direction discrimination task using moving random-dot stimuli, we found that observers' perception of the direction of motion is indeed biased away from the boundary. This misperception can be accurately described by a decoding model that preferentially weights signals from neurons whose responses best discriminate those directions. In a coarse discrimination task, to which a different decoding rule applies, the same stimulus is not misperceived, suggesting that the illusion is a direct consequence of the decoding strategy that observers use to make fine perceptual judgments. The subjective experience of motion is therefore not mediated directly by the responses of sensory neurons, but is only developed after the responses of these neurons are decoded. PMID- 17410126 TI - Template switching during break-induced replication. AB - DNA double-strand breaks (DSBs) are potentially lethal lesions that arise spontaneously during normal cellular metabolism, as a consequence of environmental genotoxins or radiation, or during programmed recombination processes. Repair of DSBs by homologous recombination generally occurs by gene conversion resulting from transfer of information from an intact donor duplex to both ends of the break site of the broken chromosome. In mitotic cells, gene conversion is rarely associated with reciprocal exchange and thus limits loss of heterozygosity for markers downstream of the site of repair and restricts potentially deleterious chromosome rearrangements. DSBs that arise by replication fork collapse or by erosion of uncapped telomeres have only one free end and are thought to repair by strand invasion into a homologous duplex DNA followed by replication to the chromosome end (break-induced replication, BIR). BIR from one of the two ends of a DSB would result in loss of heterozygosity, suggesting that BIR is suppressed when DSBs have two ends so that repair occurs by the more conservative gene conversion mechanism. Here we show that BIR can occur by several rounds of strand invasion, DNA synthesis and dissociation. We further show that chromosome rearrangements can occur during BIR if dissociation and reinvasion occur within dispersed repeated sequences. This dynamic process could function to promote gene conversion by capture of the displaced invading strand at two-ended DSBs to prevent BIR. PMID- 17410127 TI - Neuroscience: the brain's garbage men. PMID- 17410128 TI - UDP acting at P2Y6 receptors is a mediator of microglial phagocytosis. AB - Microglia, brain immune cells, engage in the clearance of dead cells or dangerous debris, which is crucial to the maintenance of brain functions. When a neighbouring cell is injured, microglia move rapidly towards it or extend a process to engulf the injured cell. Because cells release or leak ATP when they are stimulated or injured, extracellular nucleotides are thought to be involved in these events. In fact, ATP triggers a dynamic change in the motility of microglia in vitro and in vivo, a previously unrecognized mechanism underlying microglial chemotaxis; in contrast, microglial phagocytosis has received only limited attention. Here we show that microglia express the metabotropic P2Y6 receptor whose activation by endogenous agonist UDP triggers microglial phagocytosis. UDP facilitated the uptake of microspheres in a P2Y6-receptor dependent manner, which was mimicked by the leakage of endogenous UDP when hippocampal neurons were damaged by kainic acid in vivo and in vitro. In addition, systemic administration of kainic acid in rats resulted in neuronal cell death in the hippocampal CA1 and CA3 regions, where increases in messenger RNA encoding P2Y6 receptors that colocalized with activated microglia were observed. Thus, the P2Y6 receptor is upregulated when neurons are damaged, and could function as a sensor for phagocytosis by sensing diffusible UDP signals, which is a previously unknown pathophysiological function of P2 receptors in microglia. PMID- 17410129 TI - Phosphorylation of Erp1 by p90rsk is required for cytostatic factor arrest in Xenopus laevis eggs. AB - Until fertilization, the meiotic cell cycle of vertebrate eggs is arrested at metaphase of meiosis II by a cytoplasmic activity termed cytostatic factor (CSF), which causes inhibition of the anaphase-promoting complex/cyclosome (APC/C), a ubiquitin ligase that targets mitotic cyclins-regulatory proteins of meiosis and mitosis-for degradation. Recent studies indicate that Erp1/Emi2, an inhibitor protein for the APC/C, has an essential role in establishing and maintaining CSF arrest, but its relationship to Mos, a mitogen-activated protein kinase (MAPK) kinase kinase that also has an essential role in establishing CSF arrest through activation of p90 ribosomal S6 kinase (p90rsk), is unclear. Here we report that in Xenopus eggs Erp1 is a substrate of p90rsk, and that Mos-dependent phosphorylation of Erp1 by p90rsk at Thr 336, Ser 342 and Ser 344 is crucial for both stabilizing Erp1 and establishing CSF arrest in meiosis II oocytes. Semi quantitative analysis with CSF-arrested egg extracts reveals that the Mos dependent phosphorylation of Erp1 enhances, but does not generate, the activity of Erp1 that maintains metaphase arrest. Our results also suggest that Erp1 inhibits cyclin B degradation by binding the APC/C at its carboxy-terminal destruction box, and this binding is also enhanced by the Mos-dependent phosphorylation. Thus, Mos and Erp1 collaboratively establish and maintain metaphase II arrest in Xenopus eggs. The link between Mos and Erp1 provides a molecular explanation for the integral mechanism of CSF arrest in unfertilized vertebrate eggs. PMID- 17410130 TI - A direct link of the Mos-MAPK pathway to Erp1/Emi2 in meiotic arrest of Xenopus laevis eggs. AB - In vertebrates, unfertilized eggs (or mature oocytes) are arrested at metaphase of meiosis II by a cytoplasmic activity called cytostatic factor (CSF). The classical Mos-MAPK pathway has long been implicated in CSF arrest of vertebrate eggs, but exactly how it exerts CSF activity remains unclear. Recently, Erp1 (also called Emi2), an inhibitor of the anaphase-promoting complex/cyclosome (APC/C) required for degradation of the mitotic regulator cyclin B (ref. 5), has also been shown to be a component of CSF in both Xenopus and mice. Erp1 is destroyed on fertilization or egg activation, like Mos. However, despite these similarities the Mos-MAPK (mitogen-activated protein kinase) pathway and Erp1 are thought to act rather independently in CSF arrest. Here, we show that p90rsk, the kinase immediately downstream from Mos-MAPK, directly targets Erp1 for CSF arrest in Xenopus oocytes. Erp1 is synthesized immediately after meiosis I, and the Mos MAPK pathway or p90rsk is essential for CSF arrest by Erp1. p90rsk can directly phosphorylate Erp1 on Ser 335/Thr 336 both in vivo and in vitro, and upregulates both Erp1 stability and activity. Erp1 is also present in early embryos, but has little CSF activity owing, at least in part, to the absence of p90rsk activity. These results clarify the direct link of the classical Mos-MAPK pathway to Erp1 in meiotic arrest of vertebrate oocytes. PMID- 17410133 TI - Palaeontology: undressing and redressing Ediacaran embryos. AB - Bailey et al. propose that the Ediacaran microfossils Megasphaera and Parapandorina, previously interpreted as animal resting eggs and blastula embryos, represent Thiomargarita-like sulphide-oxidizing bacteria, claiming that this interpretation better explains their abundance and taphonomy. Here we highlight important observations that significantly weaken the authors' conclusions. PMID- 17410137 TI - Timber and tapirs. PMID- 17410138 TI - Museums need two cultures. PMID- 17410139 TI - South Africa's mentors. PMID- 17410141 TI - Proteins make light work of nerve control. PMID- 17410142 TI - Car emissions are EPA's problem. PMID- 17410143 TI - China plans to modernize traditional medicine. PMID- 17410145 TI - Joan of Arc's relics exposed as forgery. PMID- 17410146 TI - Smithsonian looks beyond ousted boss. PMID- 17410147 TI - Snapshot: over the moon. PMID- 17410150 TI - China's deadly drug problem. PMID- 17410153 TI - A long time ago, in a galaxy not so far away. PMID- 17410154 TI - Endangered collections. PMID- 17410155 TI - Biodiversity: logging: the new conservation. PMID- 17410156 TI - Authors defend study that shows high Iraqi death toll. PMID- 17410157 TI - Follow Thompson's map to turn biology from a science into a Science. PMID- 17410161 TI - Unity from conflict. PMID- 17410162 TI - Neuroscience: controlling neural circuits with light. PMID- 17410163 TI - Attophysics: tunnel vision. PMID- 17410164 TI - Plant biology: sticking with auxin. PMID- 17410165 TI - Palaeomagnetism: a more ancient shield. PMID- 17410166 TI - Obituary: Frank Albert Cotton (1930-2007). PMID- 17410167 TI - Attosecond real-time observation of electron tunnelling in atoms. AB - Atoms exposed to intense light lose one or more electrons and become ions. In strong fields, the process is predicted to occur via tunnelling through the binding potential that is suppressed by the light field near the peaks of its oscillations. Here we report the real-time observation of this most elementary step in strong-field interactions: light-induced electron tunnelling. The process is found to deplete atomic bound states in sharp steps lasting several hundred attoseconds. This suggests a new technique, attosecond tunnelling, for probing short-lived, transient states of atoms or molecules with high temporal resolution. The utility of attosecond tunnelling is demonstrated by capturing multi-electron excitation (shake-up) and relaxation (cascaded Auger decay) processes with subfemtosecond resolution. PMID- 17410168 TI - Multimodal fast optical interrogation of neural circuitry. AB - Our understanding of the cellular implementation of systems-level neural processes like action, thought and emotion has been limited by the availability of tools to interrogate specific classes of neural cells within intact, living brain tissue. Here we identify and develop an archaeal light-driven chloride pump (NpHR) from Natronomonas pharaonis for temporally precise optical inhibition of neural activity. NpHR allows either knockout of single action potentials, or sustained blockade of spiking. NpHR is compatible with ChR2, the previous optical excitation technology we have described, in that the two opposing probes operate at similar light powers but with well-separated action spectra. NpHR, like ChR2, functions in mammals without exogenous cofactors, and the two probes can be integrated with calcium imaging in mammalian brain tissue for bidirectional optical modulation and readout of neural activity. Likewise, NpHR and ChR2 can be targeted together to Caenorhabditis elegans muscle and cholinergic motor neurons to control locomotion bidirectionally. NpHR and ChR2 form a complete system for multimodal, high-speed, genetically targeted, all-optical interrogation of living neural circuits. PMID- 17410169 TI - Mechanism of auxin perception by the TIR1 ubiquitin ligase. AB - Auxin is a pivotal plant hormone that controls many aspects of plant growth and development. Perceived by a small family of F-box proteins including transport inhibitor response 1 (TIR1), auxin regulates gene expression by promoting SCF ubiquitin-ligase-catalysed degradation of the Aux/IAA transcription repressors, but how the TIR1 F-box protein senses and becomes activated by auxin remains unclear. Here we present the crystal structures of the Arabidopsis TIR1-ASK1 complex, free and in complexes with three different auxin compounds and an Aux/IAA substrate peptide. These structures show that the leucine-rich repeat domain of TIR1 contains an unexpected inositol hexakisphosphate co-factor and recognizes auxin and the Aux/IAA polypeptide substrate through a single surface pocket. Anchored to the base of the TIR1 pocket, auxin binds to a partially promiscuous site, which can also accommodate various auxin analogues. Docked on top of auxin, the Aux/IAA substrate peptide occupies the rest of the TIR1 pocket and completely encloses the hormone-binding site. By filling in a hydrophobic cavity at the protein interface, auxin enhances the TIR1-substrate interactions by acting as a 'molecular glue'. Our results establish the first structural model of a plant hormone receptor. PMID- 17410170 TI - Global warming and climate forcing by recent albedo changes on Mars. AB - For hundreds of years, scientists have tracked the changing appearance of Mars, first by hand drawings and later by photographs. Because of this historical record, many classical albedo patterns have long been known to shift in appearance over time. Decadal variations of the martian surface albedo are generally attributed to removal and deposition of small amounts of relatively bright dust on the surface. Large swaths of the surface (up to 56 million km2) have been observed to darken or brighten by 10 per cent or more. It is unknown, however, how these albedo changes affect wind circulation, dust transport and the feedback between these processes and the martian climate. Here we present predictions from a Mars general circulation model, indicating that the observed interannual albedo alterations strongly influence the martian environment. Results indicate enhanced wind stress in recently darkened areas and decreased wind stress in brightened areas, producing a positive feedback system in which the albedo changes strengthen the winds that generate the changes. The simulations also predict a net annual global warming of surface air temperatures by approximately 0.65 K, enhancing dust lifting by increasing the likelihood of dust devil generation. The increase in global dust lifting by both wind stress and dust devils may affect the mechanisms that trigger large dust storm initiation, a poorly understood phenomenon, unique to Mars. In addition, predicted increases in summertime air temperatures at high southern latitudes would contribute to the rapid and steady scarp retreat that has been observed in the south polar residual ice for the past four Mars years. Our results suggest that documented albedo changes affect recent climate change and large-scale weather patterns on Mars, and thus albedo variations are a necessary component of future atmospheric and climate studies. PMID- 17410171 TI - Lanthanide contraction and magnetism in the heavy rare earth elements. AB - The heavy rare earth elements crystallize into hexagonally close packed (h.c.p.) structures and share a common outer electronic configuration, differing only in the number of 4f electrons they have. These chemically inert 4f electrons set up localized magnetic moments, which are coupled via an indirect exchange interaction involving the conduction electrons. This leads to the formation of a wide variety of magnetic structures, the periodicities of which are often incommensurate with the underlying crystal lattice. Such incommensurate ordering is associated with a 'webbed' topology of the momentum space surface separating the occupied and unoccupied electron states (the Fermi surface). The shape of this surface-and hence the magnetic structure-for the heavy rare earth elements is known to depend on the ratio of the interplanar spacing c and the interatomic, intraplanar spacing a of the h.c.p. lattice. A theoretical understanding of this problem is, however, far from complete. Here, using gadolinium as a prototype for all the heavy rare earth elements, we generate a unified magnetic phase diagram, which unequivocally links the magnetic structures of the heavy rare earths to their lattice parameters. In addition to verifying the importance of the c/a ratio, we find that the atomic unit cell volume plays a separate, distinct role in determining the magnetic properties: we show that the trend from ferromagnetism to incommensurate ordering as atomic number increases is connected to the concomitant decrease in unit cell volume. This volume decrease occurs because of the so-called lanthanide contraction, where the addition of electrons to the poorly shielding 4f orbitals leads to an increase in effective nuclear charge and, correspondingly, a decrease in ionic radii. PMID- 17410172 TI - Effect of evaporite deposition on Early Cretaceous carbon and sulphur cycling. AB - The global carbon and sulphur cycles are central to our understanding of the Earth's history, because changes in the partitioning between the reduced and oxidized reservoirs of these elements are the primary control on atmospheric oxygen concentrations. In modern marine sediments, the burial rates of reduced carbon and sulphur are positively coupled, but high-resolution isotope records indicate that these rates were inversely related during the Early Cretaceous period. This inverse relationship is difficult to reconcile with our understanding of the processes that control organic matter remineralization and pyrite burial. Here we show that the inverse correlation can be explained by the deposition of evaporites during the opening of the South Atlantic Ocean basin. Evaporite deposition can alter the chemical composition of sea water, which can in turn affect the ability of sulphate-reducing bacteria to remineralize organic matter and mediate pyrite burial. We use a reaction-transport model to quantify these effects, and the resulting changes in the burial rates of carbon and sulphur, during the Early Cretaceous period. Our results indicate that deposition of the South Atlantic evaporites removed enough sulphate from the ocean temporarily to reduce biologically mediated pyrite burial and organic matter remineralization by up to fifty per cent, thus explaining the inverse relationship between the burial rates of reduced carbon and sulphur during this interval. Furthermore, our findings suggest that the effect of changing seawater sulphate concentrations on the marine subsurface biosphere may be the key to understanding other large-scale perturbations of the global carbon and sulphur cycles. PMID- 17410173 TI - Geomagnetic field strength 3.2 billion years ago recorded by single silicate crystals. AB - The strength of the Earth's early geomagnetic field is of importance for understanding the evolution of the Earth's deep interior, surface environment and atmosphere. Palaeomagnetic and palaeointensity data from rocks formed near the boundary of the Proterozoic and Archaean eons, some 2.5 Gyr ago, show many hallmarks of the more recent geomagnetic field. Reversals are recorded, palaeosecular variation data indicate a dipole-dominated morphology and available palaeointensity values are similar to those from younger rocks. The picture before 2.8 Gyr ago is much less clear. Rocks of the Archaean Kaapvaal craton (South Africa) are among the best-preserved, but even they have experienced low grade metamorphism. The variable acquisition of later magnetizations by these rocks is therefore expected, precluding use of conventional palaeointensity methods. Silicate crystals from igneous rocks, however, can contain minute magnetic inclusions capable of preserving Archaean-age magnetizations. Here we use a CO2 laser heating approach and direct-current SQUID magnetometer measurements to obtain palaeodirections and intensities from single silicate crystals that host magnetite inclusions. We find 3.2-Gyr-old field strengths that are within 50 per cent of the present-day value, indicating that a viable magnetosphere sheltered the early Earth's atmosphere from solar wind erosion. PMID- 17410174 TI - Doushantuo embryos preserved inside diapause egg cysts. AB - Phosphatized microfossils in the Ediacaran (635-542 Myr ago) Doushantuo Formation, south China, have been interpreted as the embryos of early animals. Despite experimental demonstration that embryos can be preserved, microstructural evidence that the Doushantuo remains are embryonic and an unambiguous record of fossil embryos in Lower Cambrian rocks, questions about the phylogenetic relationships of these fossils remain. Most recently, some researchers have proposed that Doushantuo microfossils may be giant sulphur-oxidizing bacteria comparable to extant Thiomargarita sp. Here we report new observations that provide a test of the bacterial hypothesis. The discovery of embryo-like Doushantuo fossils inside large, highly ornamented organic vesicles (acritarchs) indicates that these organisms were eukaryotic, and most probably early cleavage stage embryos preserved within diapause egg cysts. Large acanthomorphic microfossils of the type observed to contain fossil embryos first appear in rocks just above a 632.5 +/- 0.5-Myr-old ash bed, suggesting that at least stem-group animals inhabited shallow seas in the immediate aftermath of global Neoproterozoic glaciation. PMID- 17410175 TI - Quantifying social group evolution. AB - The rich set of interactions between individuals in society results in complex community structure, capturing highly connected circles of friends, families or professional cliques in a social network. Thanks to frequent changes in the activity and communication patterns of individuals, the associated social and communication network is subject to constant evolution. Our knowledge of the mechanisms governing the underlying community dynamics is limited, but is essential for a deeper understanding of the development and self-optimization of society as a whole. We have developed an algorithm based on clique percolation that allows us to investigate the time dependence of overlapping communities on a large scale, and thus uncover basic relationships characterizing community evolution. Our focus is on networks capturing the collaboration between scientists and the calls between mobile phone users. We find that large groups persist for longer if they are capable of dynamically altering their membership, suggesting that an ability to change the group composition results in better adaptability. The behaviour of small groups displays the opposite tendency-the condition for stability is that their composition remains unchanged. We also show that knowledge of the time commitment of members to a given community can be used for estimating the community's lifetime. These findings offer insight into the fundamental differences between the dynamics of small groups and large institutions. PMID- 17410176 TI - Antibiotic interactions that select against resistance. AB - Multidrug combinations are increasingly important in combating the spread of antibiotic-resistance in bacterial pathogens. On a broader scale, such combinations are also important in understanding microbial ecology and evolution. Although the effects of multidrug combinations on bacterial growth have been studied extensively, relatively little is known about their impact on the differential selection between sensitive and resistant bacterial populations. Normally, the presence of a drug confers an advantage on its resistant mutants in competition with the sensitive wild-type population. Here we show, by using a direct competition assay between doxycycline-resistant and doxycycline-sensitive Escherichia coli, that this differential selection can be inverted in a hyper antagonistic class of drug combinations. Used in such a combination, a drug can render the combined treatment selective against the drug's own resistance allele. Further, this inversion of selection seems largely insensitive to the underlying resistance mechanism and occurs, at sublethal concentrations, while maintaining inhibition of the wild type. These seemingly paradoxical results can be rationalized in terms of a simple geometric argument. Our findings demonstrate a previously unappreciated feature of the fitness landscape for the evolution of resistance and point to a trade-off between the effect of drug interactions on absolute potency and the relative competitive selection that they impose on emerging resistant populations. PMID- 17410177 TI - Molecular analysis of the pre-BCR complex in a large cohort of patients affected by autosomal-recessive agammaglobulinemia. AB - Autosomal-recessive agammaglobulinemia is a rare and heterogeneous disorder, characterized by early-onset infections, profound hypogammaglobulinemia of all immunoglobulin isotypes and absence of circulating B lymphocytes. To investigate the molecular basis of the disease, 23 patients with early-onset disease and no mutations in Bruton tyrosine kinase, the gene responsible for X-linked agammaglobulinemia, were selected and analyzed by direct sequencing of candidate genes. Two novel mutations in the mu heavy chain (muHC) gene (IGHM) were identified in three patients belonging to two unrelated families. A fourth patient carries a previously described G>A nucleotide substitution at the -1 position of an alternative splice site in IGHM; here, we demonstrate that this mutation is indeed responsible for aberrant splicing. Comparison of bone marrow cytofluorimetric profiles in two patients carrying different mutations in the IGHM gene suggests a genotype-phenotype correlation with the stage at which B cell development is blocked. Several new single nucleotide polymorphisms (SNPs) both in the muHC and in the lambda5-like/VpreB-coding genes were identified. Two unrelated patients carry compound heterozygous variations in the VpreB1 gene that may be involved in disease ethiology. PMID- 17410179 TI - Sensitive and precise regulation of haemoglobin after gene transfer of erythropoietin to muscle tissue using electroporation. AB - Electroporation-based gene transfer (electro gene transfer (EGT)) is gaining increasing momentum, in particular for muscle tissue, where long-term high-level expression is obtainable. Induction of expression using the Tet-On system was previously established; however, attempts to reach a predefined target dose - a prescription, have not been reported. We set three target haemoglobin levels (10, 12 and 14 mmol/l, base level was 8.2 mmol/l) and aimed at them by transferring the erythropoietin (EPO) gene to mouse tibialis cranialis (TC) muscle, and varying (1) DNA amount, (2) muscle mass transfected and (3) induction with the Tet-On system. Results showed that (a) using GFP, luciferase and EPO low DNA amounts were needed. In fact, 0.5 microg of DNA to one TC muscle led to significant Hgb elevation - this amount extrapolates to 1.4 mg of DNA in humans, (b) three prescribers hit the targets with average Hgb of 10.5, 12.0 and 13.7 mmol/l, (c) different approaches could be used, (d) undershooting could be corrected by retransferring, and (e) overshooting could be alleviated by reducing dose of inducer (doxycycline (dox)). In conclusion, this study shows that using EGT to muscle, a preset level of protein expression can be reached. This is of great interest for future clinical use. PMID- 17410180 TI - In vivo expression of GLP-1/IgG-Fc fusion protein enhances beta-cell mass and protects against streptozotocin-induced diabetes. AB - Glucagon-like peptide 1 (GLP-1) and its analogue exendin-4 (Ex4) have displayed potent glucose homeostasis-modulating characteristics in type 2 diabetes (T2D). However, there are few reports of effectiveness in type 1 diabetes (T1D) therapy, where there is massive loss of beta cells. We previously described a novel GLP-1 analogue consisting of the fusion of active GLP-1 and IgG heavy chain constant regions (GLP-1/IgG-Fc), and showed that in vivo expression of the protein, via electroporation-enhanced intramuscular plasmid-based gene transfer, normalized blood glucose levels in T2D-prone db/db mice. In the present study, GLP-1/IgG-Fc and Ex4/IgG-Fc were independently tested in multiple low-dose streptozotocin induced T1D. Both GLP-1/IgG-Fc and Ex4/IgG-Fc effectively reduced fed blood glucose levels in treated mice and ameliorated diabetes symptoms, where as control IgG-Fc had no effect. Treatment with GLP-1/IgG-Fc or Ex4/IgG-Fc improved glucose tolerance and increased circulating insulin and GLP-1 levels. It also significantly enhanced islet beta-cell mass, which is likely a major factor in the amelioration of diabetes. This suggests that GLP-1/IgG-Fc gene therapy may be applicable to diseases where there is either acute or chronic beta-cell injury. PMID- 17410181 TI - SPECT/CT imaging of baculovirus biodistribution in rat. AB - Non-invasive imaging provides essential information regarding the biodistribution of gene therapy vectors and it can also be used for the development of targeted vectors. In this study, we have utilized micro Single-photon emission computed tomography to visualize biodistribution of a (99m)Tc-polylys-ser-DTPA-biotin labelled avidin-displaying baculovirus, Baavi, after intrafemoral (i.f.), intraperitoneal (i.p.), intramuscular (i.m.) and intracerebroventricular (i.c.v.) administration. The imaging results suggest that the virus can spread via the lymphatic network after different administration routes, also showing accumulation in the nasal area after systemic administration. Extensive expression in the kidneys and spleen was seen after i.p. administration, which was confirmed by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Additionally, transduction of kidneys was seen with i.m. and i.f. administrations. We conclude that baculovirus may be beneficial for the treatment of kidney diseases after i.p. administration route. PMID- 17410184 TI - Microarray-based classification of a consecutive series of 121 childhood acute leukemias: prediction of leukemic and genetic subtype as well as of minimal residual disease status. AB - Gene expression analyses were performed on 121 consecutive childhood leukemias (87 B-lineage acute lymphoblastic leukemias (ALLs), 11 T-cell ALLs and 23 acute myeloid leukemias (AMLs)), investigated during an 8-year period at a single center. The supervised learning algorithm k-nearest neighbor was utilized to build gene expression predictors that could classify the ALLs/AMLs according to clinically important subtypes with high accuracy. Validation experiments in an independent data set verified the high prediction accuracies of our classifiers. B-lineage ALLs with uncharacteristic cytogenetic aberrations or with a normal karyotype displayed heterogeneous gene expression profiles, resulting in low prediction accuracies. Minimal residual disease status (MRD) in T-cell ALLs with a high (>0.1%) MRD at day 29 could be classified with 100% accuracy already at the time of diagnosis. In pediatric leukemias with uncharacteristic cytogenetic aberrations or with a normal karyotype, unsupervised analysis identified two novel subgroups: one consisting mainly of cases remaining in complete remission (CR) and one containing a few patients in CR and all but one of the patients who relapsed. This study of a consecutive series of childhood leukemias confirms and extends further previous reports demonstrating that global gene expression profiling provides a valuable tool for genetic and clinical classification of childhood leukemias. PMID- 17410185 TI - Complex MLL rearrangements in t(4;11) leukemia patients with absent AF4.MLL fusion allele. AB - The human mixed lineage leukemia (MLL) gene is frequently involved in genetic rearrangements with more than 55 different translocation partner genes, all associated with acute leukemia. Reciprocal chromosomal translocations generate two MLL fusion alleles, where 5'- and 3'-portions of MLL are fused to gene segments of given fusion partners. In case of t(4;11) patients, about 80% of all patients exhibit both reciprocal fusion alleles, MLL.AF4 and AF4.MLL, respectively. By contrast, 20% of all t(4;11) patients seem to encode only the MLL.AF4 fusion allele. Here, we analyzed these 'MLL.AF4(+)/AF4.MLL(-)' patients at the genomic DNA level to unravel their genetic situation. Cryptic translocations and three-way translocations were found in this group of t(4;11) patients. Reciprocal MLL fusions with novel translocation partner genes, for example NF-KB1 and RABGAP1L, were identified and actively transcribed in leukemic cells. In other patients, the reciprocal 3'-MLL gene segment was fused out-of frame to PBX1, ELF2, DSCAML1 and FXYD6. The latter rearrangements caused haploinsufficiency of genes that are normally expressed in hematopoietic cells. Finally, patients were identified that encode only solitary 3'-MLL gene segments on the reciprocal allele. Based on these data, we propose that all t(4;11) patients exhibit reciprocal MLL alleles, but due to the individual recombination events, provide different pathological disease mechanisms. PMID- 17410186 TI - CXCR4 antagonists mobilize childhood acute lymphoblastic leukemia cells into the peripheral blood and inhibit engraftment. AB - The role of CXCL12 in the bone marrow (BM) homing and growth of B-cell progenitor acute lymphoblastic leukemia (ALL) has been established. However, the effect of modulating CXCL12/CXCR4 interactions on the retention of ALL cells within the supportive BM microenvironment and the expansion and dissemination of ALL cells in vivo has not been examined. We used mouse models of human childhood and murine leukemia and specific peptide and small molecule CXCR4 antagonists to examine the importance of CXCL12/CXCR4 in the development of leukemia in vivo. CXCR4 antagonists mobilized ALL cells into the peripheral blood (PB). Extended administration of CXCR4 antagonists to mice with leukemia resulted in a reduction in the number of leukemic cells in the PB and spleens of animals compared to control treated animals in three of the five cases tested. There was also a marked reduction in the dissemination of ALL cells to extramedullary sites including liver and kidney in all cases where this occurred. Considering the inhibitory effect of stromal layers on the activity of chemotherapeutic agents and the interactive effect of CXCL12 antagonists with chemotherapeutic agents in vitro, this raises the possibility of using these agents to potentiate the effects of current chemotherapy regimens. PMID- 17410187 TI - Mechanisms of action of antithymocyte globulin: T-cell depletion and beyond. AB - The success of allogeneic stem cell transplantation and solid-organ transplantation owes much to improvements in the immunosuppressive regimens that prevent graft-versus-host disease (GVHD) or suppress allograft rejection. A better understanding of the immune mechanisms underlying induction of immunological tolerance is the key to successful transplantation. Polyclonal antibodies such as antithymocyte globulins (ATG) have been used for decades. The common belief is that ATG efficacy relies on its capacity to deplete T lymphocytes. The aim of this review is to offer an overview of the recent findings that have been demonstrated in ATG's immunomodulatory activity. The polyclonal nature of ATG is reflected in its diverse effects on the immune system: (1) T-cell depletion in blood and peripheral lymphoid tissues through complement-dependent lysis and T-cell activation and apoptosis; (2) modulation of key cell surface molecules that mediate leukocyte/endothelium interactions; (3) induction of apoptosis in B-cell lineages; (4) interference with dendritic cell functional properties; and (5) induction of regulatory T and natural killer T cells. As a consequence, ATG provides multifaceted immunomodulation paving the way for future applications and suggesting that the use of ATG should be included in the immunosuppression therapeutic armamentarium to help reduce the incidence of organ rejection and GVHD. PMID- 17410188 TI - A different schedule of zoledronic acid can reduce the risk of the osteonecrosis of the jaw in patients with multiple myeloma. AB - Osteonecrosis of the jaw (ONJ) is a reported complication of bisphosphonate use. The incidence ranges between 6 and 13% and seems to be higher in people treated with zoledronic acid (ZA) than with pamidronate. We retrospectively evaluated the incidences of ONJ and skeletal-related events (SRE) in 106 patients with multiple myeloma divided in two groups according to the schedule of administration of bisphosphonates: 51 received monthly administrations until tolerated (group A, standard schedule), 55 were treated monthly during the first year and then every 3 months (group B, reduced schedule). The incidence of SRE was similar (15.1 per 100 person years in group A and 17.7 in group B). ONJ occurred in seven patients, six in group A and one in group B (P=0.049). The risk of ONJ was eight-fold lower with the reduced schedule than with the standard schedule. The only significant risk factor for ONJ was the type of bisphosphonate (P=0.006). The incidence of ONJ was significantly higher with ZA than with pamidronate + ZA (9.1 vs 1.6 per 100 person-years). No ONJ was observed in patients treated only with pamidronate. A reduced schedule of ZA may be safer than the standard schedule while maintaining anti-resorptive efficacy. PMID- 17410189 TI - Quantitative profiling of codon 816 KIT mutations can aid in the classification of systemic mast cell disease. PMID- 17410190 TI - A new type of NPM1 gene mutation in AML leading to a C-terminal truncated protein. PMID- 17410191 TI - Small number of HTLV-1-positive cells frequently remains during complete remission after allogeneic hematopoietic stem cell transplantation that are heterogeneous in origin among cases with adult T-cell leukemia/lymphoma. AB - Allogeneic hematopoietic stem cell transplantation (allo-HSCT) can provide long term remission for patients with adult T-cell leukemia/lymphoma (ATLL) caused by human retrovirus, human T-lymphocyte virus (HTLV-1). To understand how HTLV-1 positive cells including ATLL cells were suppressed by allo-HSCT, we examined HTLV-1 provirus load and residual ATLL cells in peripheral blood of transplant recipients using PCR-based tests. We found that the copy number of HTLV-1 genome, called provirus, became very small in number after allo-HSCT; however, in most cases, provirus did not disappear even among long-term survivors. Tumor-specific PCR tests demonstrated that most of HTLV-1-positive cells that remained long after transplantation were not primary ATLL cells but donor-derived HTLV-1 positive cells. We also found a case having very low amount of residual disease in peripheral blood even long after transplantation. There was only one recipient in whom we failed to show the presence of HTLV-1 genome and antibody against HTLV 1 even with an extensive search, which strongly suggested the elimination of HTLV 1 after allo-HSCT. These results demonstrated that after allo-HSCT the small amount of residual HTLV-1-positive cells were heterogeneous in origin and that long-term disease control for ATLL could be obtained without the complete elimination of HTLV-1. PMID- 17410192 TI - Discriminant function analysis as decision support system for the diagnosis of acute leukemia with a minimal four color screening panel and multiparameter flow cytometry immunophenotyping. AB - Despite several recommendations for standardization of multiparameter flow cytometry (MFC) the number, specificity and combinations of reagents used by diagnostic laboratories for the diagnosis and classification of acute leukemias (AL) are still very diverse. Furthermore, the current diagnostic interpretation of flow cytometry readouts is influenced arbitrarily by individual experience and knowledge. We determined the potential value of a minimal four-color combination panel of 13 monoclonal antibodies (mAbs) with a CD45/sideward light scatter gating strategy for a standardized MFC immunophenotyping of the clinically most relevant subgroups of AL. Bone marrow samples from 155 patients with acute myeloid leukemia (AML, n=79), B-cell precursor acute lymphoblastic leukemia (BCP ALL, n=29), T-cell precursor acute lymphoblastic leukemia (T-ALL, n=12) and normal bone marrow donors (NBMD, n=35) were analyzed. A knowledge-based learning algorithm was generated by comparing the results of the minimal panel with the actual diagnosis, using discriminative function analysis. Correct classification of the test sample according to lineage, that is, BCP-ALL, T-ALL, AML and differentiation of NBMD was achieved in 97.2% of all cases with only six of the originally applied 13 mAbs of the panel. This provides evidence that discriminant function analysis can be utilized as a decision support system for interpretation of flow cytometry readouts. PMID- 17410193 TI - Class I HDAC SNP analysis in healthy donors compared to AML patients. PMID- 17410194 TI - Germline mutations in SPI1 and MADD do not contribute to familial chronic lymphocytic leukaemia. PMID- 17410195 TI - Gene expression overlap affects karyotype prediction in pediatric acute lymphoblastic leukemia. PMID- 17410196 TI - Preformed metal crowns may last longer than fillings. AB - DATA SOURCES: Searches were made using the Cochrane Central Register of Controlled Trials (CENTRAL), Medline, Embase and the System for Information on Grey Literature in Europe (SIGLE). STUDY SELECTION: Randomised controlled trials (RCT) were chosen that assessed the effectiveness of PMC, compared with filling materials or in children where there had been no treatment of tooth decay in one or more primary molar teeth. DATA EXTRACTION AND SYNTHESIS: Forty-seven records were retrieved by the search strategies, some of which proved to be duplicates. Ultimately, 14 studies were scrutinised. None met the inclusion criteria and six studies were excluded from the review because they were either retrospective in design or they reported prospective outcomes but were not randomised. CONCLUSIONS: No RCT were available for appraisal. Although the use of PMC is recommended in clinical practice by the British Society of Paediatric Dentistry (BSPD), the evidence to support this is not strong, consisting mainly of case reports and uncontrolled studies. The lower quality levels of evidence that have been produced nevertheless have some strength, since clinical outcomes are consistently in favour of PMC even though many studies analyse casts placed on the most damaged of the pair of teeth. It is important that the absence of evidence for PMC is not misinterpreted as evidence for their lack of efficacy. PMID- 17410197 TI - Filaggrin null alleles are not associated with psoriasis. AB - Psoriasis is a common skin disease with an etiology consistent with a multifactorial trait. Several psoriasis susceptibility loci are known, a number of which are also implicated in a predisposition to atopic dermatitis (AD), including the epidermal differentiation complex on chromosome 1q21. It has recently been shown in several replicate studies that prevalent null alleles for the filaggrin gene (FLG) on 1q21 are an important genetic factor in AD. Here, we examined the role of these FLG variants in psoriasis using case:control association studies comparing Irish and UK psoriasis cohorts (combined n=691) to ethnically matched populations (combined n=2117). No association was present for the two common European FLG mutations R501X and 2282del4 (combined chi2 P=0.989). In addition, the 3' end of the FLG open-reading frame was sequenced in a number of patients with differing types of psoriasis (plaque, guttate, palmoplantar, and late-onset), which excluded the possibility of a gain-of-function frameshift mutation such as those found in loricrin or certain keratin genes. These data suggest that FLG mutations are unlikely to be involved in genetic susceptibility to psoriasis and implies that there may be within-locus heterogeneity in chromosomal regions involved in both AD and psoriasis. PMID- 17410198 TI - Polymorphisms in folate, pyrimidine, and purine metabolism are associated with efficacy and toxicity of methotrexate in psoriasis. AB - Methotrexate is the gold standard therapy for moderate to severe psoriasis, but there is marked interpersonal variation in its efficacy and toxicity. We hypothesized that in psoriasis patients, specific common polymorphisms in folate, pyrimidine, and purine metabolic enzymes are associated with methotrexate efficacy and/or toxicity. DNA from 203 retrospectively recruited psoriasis patients treated with methotrexate was collected and genotyped by restriction endonuclease digestion or length polymorphism assays. The reduced folate carrier (RFC) 80A allele and the thymidylate synthase (TS) 3'-untranslated region (3' UTR) 6 bp deletion were associated with methotrexate-induced toxicity (P=0.025 and P=0.025, respectively). RFC 80A and 5-aminoimidazole-4-carboxamide ribonucleotide transformylase (ATIC) 347G were associated with methotrexate discontinuation (P=0.048 and P=0.038). The TS 5'-UTR 28 bp 3R polymorphism correlated with poor clinical outcome (P=0.029), however, this was not the case when patients with palmoplantar pustular psoriasis were not included in the analysis. Stronger associations between specific polymorphisms and methotrexate induced toxicity and discontinuation were found in a subanalysis of patients on methotrexate not receiving folic acid supplementation. We have demonstrated preliminary evidence that specific polymorphisms of enzymes involved in folate, pyrimidine, and purine metabolism could be useful in predicting clinical response to methotrexate in patients with psoriasis. PMID- 17410199 TI - P75 plays a key role in the induction of the sprouting of sensory nerve fibers in inflamed skin. PMID- 17410200 TI - Low concentrations of curcumin induce growth arrest and apoptosis in skin keratinocytes only in combination with UVA or visible light. AB - It is well known that curcumin, a dietary pigment from the plant Curcuma longa, inhibits cell proliferation and induces apoptosis in different cell lines at concentrations ranging from 10 to 150 microM (3.7-55 microg/ml). In this study, we show that curcumin at low concentrations (0.2-1 microg/ml) also has an antiproliferative effect when applied in combination with UVA or visible light. We demonstrate that such a treatment induces apoptosis in human skin keratinocytes represented by the increase of fragmented cell nuclei, release of cytochrome c from mitochondria, activation of caspases-9 and -8, and inhibition of NF-kappaB activity. Furthermore, inhibition of extracellular regulated kinases 1/2 and protein kinase B was found to ensure the proapoptotic effect. Additionally, the EGFR, an upstream regulator of both kinases, was inhibited indicating that apoptosis is induced by blocking survival- and proliferation associated signal cascades at the receptor level. In summary, these findings suggest a new therapeutic concept for the treatment of hyperproliferative diseases by combining topical curcumin with UVA or visible light. In particular, the latter avoids the use of carcinogenic irradiation that is part of regular phototherapy. PMID- 17410201 TI - Rodent Lce gene clusters; new nomenclature, gene organization, and divergence of human and rodent genes. PMID- 17410202 TI - PIN1, the cell cycle and cancer. AB - PIN1 is a peptidyl-prolyl isomerase that can alter the conformation of phosphoproteins and so affect protein function and/or stability. PIN1 regulates a number of proteins important for cell-cycle progression and, based on gain- and loss-of-function studies, is presumed to operate as a molecular timer of this important process. Therefore, it seems logical that alterations in the level of PIN1 can influence hyperproliferative diseases such as cancer. However, the precise role of PIN1 in cancer remains controversial. PMID- 17410203 TI - Cajal bodies and the nucleolus are required for a plant virus systemic infection. AB - The nucleolus and Cajal bodies (CBs) are prominent interacting subnuclear domains involved in a number of crucial aspects of cell function. Certain viruses interact with these compartments but the functions of such interactions are largely uncharacterized. Here, we show that the ability of the groundnut rosette virus open reading frame (ORF) 3 protein to move viral RNA long distances through the phloem strictly depends on its interaction with CBs and the nucleolus. The ORF3 protein targets and reorganizes CBs into multiple CB-like structures and then enters the nucleolus by causing fusion of these structures with the nucleolus. The nucleolar localization of the ORF3 protein is essential for subsequent formation of viral ribonucleoprotein (RNP) particles capable of virus long-distance movement and systemic infection. We provide a model whereby the ORF3 protein utilizes trafficking pathways involving CBs to enter the nucleolus and, along with fibrillarin, exit the nucleus to form viral 'transport-competent' RNP particles in the cytoplasm. PMID- 17410204 TI - The morphology proteins Mdm12/Mmm1 function in the major beta-barrel assembly pathway of mitochondria. AB - The beta-barrel proteins of mitochondria are synthesized on cytosolic ribosomes. The proteins are imported by the translocase of the outer membrane (TOM) and the sorting and assembly machinery (SAM). It has been assumed that the SAM(core) complex with the subunits Sam35, Sam37 and Sam50 represents the last import stage common to all beta-barrel proteins, followed by splitting in a Tom40-specific route and a route for other beta-barrel proteins. We have identified new components of the beta-barrel assembly machinery and show that the major beta barrel pathway extends beyond SAM(core). Mdm12/Mmm1 function after SAM(core) yet before splitting of the major pathway. Mdm12/Mmm1 have been known for their role in maintenance of mitochondrial morphology but we reveal assembly of beta-barrel proteins as their primary function. Moreover, Mdm10, which functions in the Tom40 specific route, can associate with SAM(core) as well as Mdm12/Mmm1 to form distinct assembly complexes, indicating a dynamic exchange between the machineries governing mitochondrial beta-barrel assembly. We conclude that assembly of mitochondrial beta-barrel proteins represents a major function of the morphology proteins Mdm12/Mmm1. PMID- 17410205 TI - The restriction fold turns to the dark side: a bacterial homing endonuclease with a PD-(D/E)-XK motif. AB - The homing endonuclease I-Ssp6803I causes the insertion of a group I intron into a bacterial tRNA gene-the only example of an invasive mobile intron within a bacterial genome. Using a computational fold prediction, mutagenic screen and crystal structure determination, we demonstrate that this protein is a tetrameric PD-(D/E)-XK endonuclease - a fold normally used to protect a bacterial genome from invading DNA through the action of restriction endonucleases. I-Ssp6803I uses its tetrameric assembly to promote recognition of a single long target site, whereas restriction endonuclease tetramers facilitate cooperative binding and cleavage of two short sites. The limited use of the PD-(D/E)-XK nucleases by mobile introns stands in contrast to their frequent use of LAGLIDADG and HNH endonucleases - which in turn, are rarely incorporated into restriction/modification systems. PMID- 17410206 TI - Germline transcription from T-cell receptor Vbeta gene is uncoupled from allelic exclusion. AB - Allelic exclusion operates in B and T lymphocytes to ensure clonal expression of antigen receptors after V(D)J recombination. Germline transcription, which proceeds V(D)J recombination, has been postulated to provide an instructive signal for allelic exclusion. Here, we use a genetic marker to track germline transcription from a Vbeta gene within the TCRbeta locus. We find that developing thymocytes exhibit uniformed, bi-allelic activation of the Vbeta gene before V-DJ recombination, a process subject to allelic exclusion. We further show that V-DJ rearrangement promotes activation rather than silencing of germline transcription from the remaining Vbeta genes on either the functionally or non-functionally rearranged chromosome. Results presented here suggest that germline transcription, although necessary for V(D)J recombination, is not sufficient to instruct allelic exclusion. PMID- 17410207 TI - A novel role for histone chaperones CAF-1 and Rtt106p in heterochromatin silencing. AB - The histone chaperones CAF-1 and Rtt106p are required for heterochromatin silencing in the yeast Saccharomyces cerevisiae. Although it has been suggested that CAF-1 is involved in the maintenance of heterochromatin silencing, their exact functions during this process are not well understood. Here, we show that CAF-1 and Rtt106p are involved in the early stages of heterochromatin formation. The binding of Sir proteins to telomeric heterochromatin is significantly reduced and, additionally, Sir proteins are mislocalized in cells lacking CAF-1 and Rtt106p. At the HMR locus, CAF-1 and Rtt106p are required for the initial recruitment of Sir2p and Sir3p, but not Sir4p, to the HMR-E silencer, where silencing initiates, as well as the efficient spreading of all of these Sir proteins to the distal a1 gene. Moreover, silencing at the HMR locus is dramatically reduced in cells lacking CAF-1, Rtt106p, and Sir1p. Thus, these studies reveal a novel role for CAF-1 and Rtt106p in epigenetic silencing and indicate that the spreading of heterochromatin, a poorly understood process, requires histone chaperones. PMID- 17410208 TI - Dissecting mechanisms of nuclear mRNA surveillance in THO/sub2 complex mutants. AB - The nuclear exosome is involved in numerous RNA metabolic processes. Exosome degradation of rRNA, snoRNA, snRNA and tRNA in Saccharomyces cerevisiae is activated by TRAMP complexes, containing either the Trf4p or Trf5p poly(A) polymerase. These enzymes are presumed to facilitate exosome access by appending oligo(A)-tails onto structured substrates. Another role of the nuclear exosome is that of mRNA surveillance. In strains harboring a mutated THO/Sub2p system, involved in messenger ribonucleoprotein particle biogenesis and nuclear export, the exosome-associated 3' --> 5' exonuclease Rrp6p is required for both retention and degradation of nuclear restricted mRNAs. We show here that Trf4p, in the context of TRAMP, is an mRNA surveillance factor. However, unlike Rrp6p, Trf4p only partakes in RNA degradation and not in transcript retention. Surprisingly, a polyadenylation-defective Trf4p protein is fully active, suggesting polyadenylation-independent mRNA degradation. Transcription pulse-chase experiments show that HSP104 molecules undergoing quality control in THO/sub2 mutant strains fall into two distinct populations: One that is quickly degraded after transcription induction and another that escapes rapid decay and accumulates in foci associated with the HSP104 transcription site. PMID- 17410209 TI - CBP/p300 are bimodal regulators of Wnt signaling. AB - Many Wnts influence cell behavior by a conserved signaling cascade that promotes the stabilization and nuclear accumulation of beta-catenin (beta-cat), which then associates with TCF family members to activate target genes. The histone acetyltransferase CREB binding protein (CBP) can bind to TCF and inhibit Wnt signaling in Drosophila. In contrast, studies in vertebrates indicate a positive role for CBP and the closely related protein p300 as beta-cat binding transcriptional co-activators. We address this discrepancy by demonstrating that in addition to its negative role, CBP has an essential positive role in Wnt signaling in flies. CBP binds directly to the C-terminus of Armadillo (Arm, the fly beta-cat) and is recruited to a Wnt-regulated enhancer (WRE) in a Wnt- and Arm-dependent manner. In a human colorectal cancer cell line, we show that CBP and p300 can inhibit Wnt signaling and demonstrate that human p300 can bind directly to TCF4 in vitro. Our results argue that CBP/p300 has an evolutionarily conserved role as a buffer regulating TCF-beta-cat/Arm binding. Subsequent to this interaction, it also has an essential role in mediating the transactivation activity of beta-cat/Arm. PMID- 17410210 TI - DNA damage recognition and repair by 3-methyladenine DNA glycosylase I (TAG). AB - DNA glycosylases help maintain the genome by excising chemically modified bases from DNA. Escherichia coli 3-methyladenine DNA glycosylase I (TAG) specifically catalyzes the removal of the cytotoxic lesion 3-methyladenine (3mA). The molecular basis for the enzymatic recognition and removal of 3mA from DNA is currently a matter of speculation, in part owing to the lack of a structure of a 3mA-specific glycosylase bound to damaged DNA. Here, high-resolution crystal structures of Salmonella typhi TAG in the unliganded form and in a ternary product complex with abasic DNA and 3mA nucleobase are presented. Despite its structural similarity to the helix-hairpin-helix superfamily of DNA glycosylases, TAG has evolved a modified strategy for engaging damaged DNA. In contrast to other glycosylase-DNA structures, the abasic ribose is not flipped into the TAG active site. This is the first structural demonstration that conformational relaxation must occur in the DNA upon base hydrolysis. Together with mutational studies of TAG enzymatic activity, these data provide a model for the specific recognition and hydrolysis of 3mA from DNA. PMID- 17410211 TI - Birth and hatching: key events in the onset of awareness in the lamb and chick. AB - The neuroanatomical and neurophysiological development of the embryo and fetus and unique features of the physiological environment of the fetal brain, features which are lost at birth, support recent conclusions that under normal circumstances awareness (or consciousness) is probably not exhibited by the ovine embryo-fetus before birth and that it appears for the first time only after birth. However, there has apparently been no evaluation of whether or not similar mechanisms modulate awareness-related functions in domestic chicks before and after hatching. This comparative review, in seeking to rectify this, arrived at the following conclusions. First, the neural apparatus of both lambs and chicks appears to be too immature to support any states resembling awareness during at least the first half of pregnancy or incubation. Second, electroencephalographic (EEG) activity, which evolves subsequently, shows that states of sleep-like unconsciousness are likely to be continuously present in lambs until after birth, and that such states at least predominate in chicks until after hatching. Third, as in fetal lambs, epochs of so-called 'wakefulness' previously reported in chick embryos do not seem likely to represent short periods of awareness in ovo. Fourth, several neurosuppressive mechanisms, with some unique features, also operate or have the potential to operate in chicks before hatching, but a dearth of published information currently hinders a full comparison with those demonstrated to operate in fetal lambs. Fifth, contradicting the intuitive perception that vocalisation pre-hatching by the chick indicates the presence of awareness, published evidence suggests that vocalisation before and during hatching occurs mostly during EEG states indicating sleep-like unconsciousness. Sixth, as seems to be the case for newborn lambs after birth, it is possible that demonstrable awareness may appear for the first time only after hatching in chicks, presumably through waning neurosuppression and burgeoning neuroactivation, but such awareness seems to take longer to manifest itself. However, additional research in chicks is recommended to further assess this suggestion. Particular attention should be given to the status of vocal interactions between hen and chick which begin several days before hatching, and to the operation of neurosuppressive and neuroactivating mechanisms throughout the last 40% of incubation and during and after hatching. PMID- 17410212 TI - Public attitudes in New Zealand towards the use of animals for research, testing and teaching purposes. AB - AIM: To investigate the awareness of and interest in the use of animals for research, testing and teaching (RTT) purposes, the levels of support and attitudes towards this use of animals, and the awareness of and opinions on regulations governing it within the public in New Zealand. METHODS: An independent telephone survey was used to collect information including demographics, awareness and interest in the use of animals for RTT, attitudes towards the use of animals for RTT, and awareness of the regulation of the use of animals in RTT and the degree of confidence held in such regulation. RESULTS: Data were obtained from 750 respondents throughout New Zealand. The rim-weighting efficiency figure was 90%, indicating that the sample collected closely matched the population demographics for New Zealand. Overall, 33% of respondents expressed an interest in the issue generally, 39% were interested in the area of animals used for research and testing purposes, and 21% for teaching purposes. A majority of respondents agreed that the use of animals for teaching (72%) and research and testing (68%) purposes was acceptable as long as there was no unnecessary suffering by the animals. Respondents felt animal research was most justified for research into life-threatening and debilitating diseases, and least justified for safety-testing of cosmetics and household chemicals. Only 8% of respondents knew a fair amount (or more) about the legislation pertaining to the use of animals in RTT. CONCLUSIONS: The majority of New Zealanders were neither interested nor concerned about the use of animals in RTT. The largest proportion of respondents fell into a category of accepting this use of animals with conditions such as where there is no unnecessary suffering by the animals or where the research is for a serious disease. However, the responses also indicated that attitudes towards the use of animals in RTT as a general issue may change when that use is given a specific context. CLINICAL RELEVANCE: The understanding of public opinion with regard to the use of animals in RTT is an important tool for New Zealand Veterinary Association nominees on animal ethics committees (AECs), for these veterinarians, as well as acting as assessors of the welfare of the animals involved, are also independent of the institutions using animals, so that, along with the other 'external' members, they provide a societal perspective to the deliberations of AECs. PMID- 17410213 TI - Effects of application in spring of urea fertiliser on aspects of reproductive performance of pasture-fed dairy cows. AB - AIMS: To assess if raising concentrations of crude protein (CP) in pasture in spring by the frequent application of urea fertiliser would affect ovarian follicular dynamics, luteal function, onset of oestrus and reproductive performance of dairy cows under farming conditions in New Zealand. METHODS: Spring-calved dairy cows were grazed for 101 days in paddocks that were either not fertilised (Control; n=20) during the course of the study, or were fertilised with 40-50 kg nitrogen (N)/ha every 4-6 weeks (High-N; n=20). Similar generous pasture allowances were offered to both groups. Concentrations of CP in pasture, urea in serum and progesterone in milk were measured. Ovarian follicular and luteal dynamics were determined using ultrasonography. Oestrous behaviour and the number, time and outcome of inseminations were also recorded. RESULTS: Mean concentrations of CP in pasture and urea in serum was higher in the High-N than the Control group (25.2 vs 21.6 and 8.3 vs 5.4 mmol/L for CP and urea, respectively; p<0.001). Intervals between calving and first oestrus, first insemination and conception, the time of first emergence of a dominant follicle, milk progesterone concentration, and the diameter of the corpus luteum (CL) in the first luteal phase did not differ significantly between groups. The interval from calving to first ovulation tended (p=0.10) to be lower and the diameter of the dominant follicle of the oestrous cycle at which cows conceived was greater (p=0.02) in Control than High-N cows. CONCLUSIONS: The use of large amounts of urea fertiliser during spring and the consequent increases in concentrations of CP in pasture and urea in serum did not negatively affect any of the parameters of reproductive performance of pasture-fed dairy cows that were assessed in this study. PMID- 17410214 TI - The effect of a temporal change in ingestion rates of fluorine (F) in soil on the concentration of F in serum and bone of young sheep. AB - AIM: To determine the changes in concentrations of fluorine (F) in serum and bone of young sheep given a high intake of F, as F in soil, followed by a low intake of F, to simulate the varying intakes experienced by grazing sheep. METHODS: Sixty autumn-born, weaned, mixed-sex lambs were randomly divided into two groups (n=30). One group (Control) was fed a low-F (12 mg/kg dry matter; DM) lucerne/ grain diet for 345 days, while the other group (High soil-F) was fed a high-F (224 mg/kg DM) lucerne/grain diet containing 10% soil for 94 days and then a low F diet for a further 251 days. Blood samples were collected on Days 1, 32, 94, 108, 153, 214, 280 and 345, to determine concentrations of F in serum. Groups of five or six animals from each group were killed on Days 1, 94, 153, 214, 280 and 345, to collect radii and humeri for determination of F content. The sheep were weighed at about 6-8-weekly intervals. RESULTS: Initial liveweight of the sheep was 22.1 (standard error (SE) 1.12) kg and the mean liveweight gains were 270 (SE 22.1) and 170 (SE 12.4) g/day from Days 1 to 94 and 95 to 330, respectively. The mean initial (Day 1) concentration of F in serum was 0.03 (SE 0.005) mg/L, and this changed very little for animals on the low-F diet. In sheep on the high soil F diet, the concentration of F in serum reached 0.38 (SE 0.021) mg/L at Day 94 but after being placed on the low-F diet concentrations decreased to 0.04 (SE 0.006) mg/L after 14 days (Day 108) and then remained at about this low level for the remainder of the duration of the study. The mean concentrations of F in the humerus and radius of the lambs at Day 1 were 161 (SE 19.4) and 159 (SE 20.4) mg/kg DM, respectively, which increased to 2,784 (SE 80.6) and 1,805 (SE 51.5) mg/kg DM, respectively, at Day 94 in lambs fed the high soil-F diet, then decreased to 1,075 (SE 58.1) and 1,064 (SE 61.6) mg/kg DM at Day 153, and then progressively increased to 1,669 (SE 57.7) and 1,312 (SE 30.8) mg/kg DM at Day 345. CONCLUSIONS: The ingestion of F in soil by sheep markedly increased their concentrations of F in serum and bone, but when the soil was removed and they were fed a low-F diet, concentrations of F in serum decreased to baseline levels within 14 days while concentrations in the bone only decreased to 60-70% of the peak concentration, indicating that some of the F in bone was remobilised. PMID- 17410215 TI - Efficacy, distribution and faecal excretion of copper oxide wire particles in a novel bolus in red deer (Cervus elaphus). AB - AIM: To determine the efficacy of a novel copper oxide wire particle (COWP) formulation in elevating concentrations of copper (Cu) in the liver and serum of red deer (Cervus elaphus), and to investigate the distribution of particles in the gastrointestinal tract and the rate of their excretion in faeces. METHODS: Mixed-age red deer hinds were allocated to three groups (n=10 per group) on the basis of pre-treatment liver Cu concentrations. Groups 1 and 2 were treated orally with a 10-g COWP bolus on Days 0 and 30, respectively, while the remaining group served as an untreated control. Animals were slaughtered on Day 60, when blood and liver samples were collected for determination of Cu concentrations. An additional group of 18-month-old red deer hinds (n=20) were treated orally with a 10-g COWP bolus, and four were slaughtered on each of Days 1, 5, 15, 30 and 60 after treatment. The gastrointestinal tract was secured between compartments below the oesophagus and contents rinsed until sedimentation of particles was complete. The sediment was oven-dried and COWP were separated and weighed. Faeces were collected continuously from four additional animals held in metabolism cages for 4 days after treatment, sub-sampled daily, and COWP recovered. RESULTS: Mean liver Cu concentrations at slaughter were 80, 597 and 447 micromol/kg for controls and hinds treated 30, and 60 days previously, respectively. Corresponding mean serum Cu concentrations were 7.7, 12.9 and 11.9 micromol/L, respectively. Liver and serum Cu concentrations were higher in both treatment groups than in untreated control animals (p<0.001). COWP were found in all compartments of the gastrointestinal tract measured, for at least 15 days, and in the rumen/reticulum and abomasum for at least 60 days post-administration. The highest rate of recovery overall was from the rumen/reticulum. Mean weight of COWP recovered from faeces was 0.09 g during the first 24 h and 0.94 g over the first 4 days following administration. CONCLUSION: The COWP bolus tested resulted in elevated mean liver Cu concentrations for at least 60 days compared with control animals. The majority of COWP were found in the rumen/ reticulum, where recovery was possible for at least 60 days. About 10% of particle weight was excreted in the faeces within 4 days of administration. CLINICAL SIGNIFICANCE: The test bolus was efficacious in deer, elevating mean liver and serum Cu concentrations 30 and 60 days after treatment. Variation in faecal excretion may explain between-animal differences in efficacy. PMID- 17410216 TI - A survey of anthelmintic use and internal parasite control in farmed deer in New Zealand. AB - AIM: To survey parasite control programmes and anthelmintic usage over a 12-month period on deer farms in New Zealand. METHODS: A questionnaire of general farm data, policy and procedures for anthelmintic use, anthelmintic programmes for weaner and older deer, general information and understanding of parasite management practices was posted to 500 deer farmers in November 2004. RESULTS: Two hundred and twenty-seven (45.4%) replies were received, and 198 (39.6%) were suitable for analysis. Ninety four percent of respondents used anthelmintics at least once in the 12-month period; 53% treated all classes of deer and 22% treated only weaners. Seventy-four percent based anthelmintic dose on weight of the heaviest animal, and 36% used a weigh scale. Weaner deer (n=175 farms) were treated 1-13 (mean 3.2) times in their first 12 months, at the earliest commencing in January and at the latest in November. The mean interval between treatments ranged from 41 to 46 days. Yearling and adult hinds and stags were treated at least once (range 1-7 times) on 55-64% of farms, depending on class of animal. Moxidectin was the anthelmintic most commonly used (46-58%, depending on class of animal), followed by abamectin, eprinomectin, oxfendazole, ivermectin, albendazole, levamisole and doramectin. Perceived efficacy was the most common reason for choosing a type of anthelmintic. Weight gain and body condition were the most common measures used for monitoring parasitism in weaners and older deer, respectively, and few respondents used faecal egg and/or larval counts. Coughing and/or scouring were associated with parasitism in weaners on 13-14% of farms, and deaths associated with lungworm and gastrointestinal parasites were recorded on 5% and 3% of farms, respectively. Veterinary input to diagnosis was involved in 23% of events. Production losses and/or death of yearling and/or adult deer due to parasitism were reported by 27% of respondents. When planning anthelmintic treatment programmes, 63% of respondents followed advice from veterinarians. Thirty four percent always placed deer on clean or spelled pastures after treatment, while 32% did that often. Fifteen percent had incorporated forages and/or herbs with assumed anthelmintic properties into their parasite control programme. Forty-four percent were very confident of a return on investment when using anthelmintics for their deer. Respondents stated that their knowledge of the life cycle of the major parasites of deer was very good (8%), reasonably good (61%), poor (28%), or nil (3%). CONCLUSION: Anthelmintics are used on almost all deer farms, and practices and programmes vary considerably. Opportunities exist for veterinarians to assist deer farmer clients to improve their parasite control programmes. PMID- 17410217 TI - Outcome of lag-screw treatment of incomplete fractures of the frontal plane of the radial facet of the third carpal bone in horses. AB - AIM: To describe outcomes for horses diagnosed with incomplete, non-displaced fractures of the frontal plane of the radial facet (INFR) of the third carpal bone (C3) treated by placement of a lag screw across the fracture under arthroscopic guidance. METHODS: Horses (n=13) diagnosed with INFR and treated between December 1999 and January 2005 using a lag screw placed over the fracture were studied. For each case, five horses matched for sire, age and sex which were not known to have INFR were sought for comparison. Racing performance data were collected from a commercial online database. The racing performance of cases pre- and post-operatively, and of cases and matched horses in the post-operative period was compared. RESULTS: Sixteen INFR were found in the 13 horses. Radiographic evidence of healed fracture lines 2-4 months after surgery was seen in 11/16 (69%) fractures; 11/13 (85%) cases raced again after a median recovery period of 292 (range 149-681) days. Treatment was considered successful in 9/13 (69%) cases, which were still in training or had been retired for reasons other than lameness localised to the middle carpal joint at the end of the study period. Just 6/13 (46%) cases had raced prior to injury. The racing ability pre- and post-operatively of five cases was compared, three (60%) of which performed better post-operatively than they had before. There was no significant difference in racing longevity or ability post-operatively between patients and matched (control) horses. CONCLUSIONS: Post-operatively, there was little difference in the racing performance between horses diagnosed with INFR which had a lag screw placed across the fracture line and horses matched for sire, age and sex which were not known to have INFR. CLINICAL RELEVANCE: Horses which were diagnosed with INFR of C3 and had a lag screw placed across the fracture had a good prognosis for future racing performance. PMID- 17410218 TI - How do you mean? The case for composite faecal egg counts in testing for drench resistance. PMID- 17410219 TI - Definitions of the degree of polarization of a light beam. AB - A necessary and sufficient condition is derived for certain ad hoc expressions that are frequently used in the literature to represent correctly the degree of polarization of a light beam. PMID- 17410220 TI - Investigation on spectral loss characteristics of subwavelength terahertz fibers. AB - By measuring the spectral loss characteristics of subwavelength-diameter terahertz fibers, our study supports the recent theory proposed by M. Sumetsky [Opt. Lett. 31, 870 (2006)] that diameter-variation-induced radiation is a dominant loss mechanism for subwavelength fibers in the low- (<1%) core-fraction power regime. This physical mechanism limits the lowest guidable frequency in a subwavelength fiber. PMID- 17410221 TI - Photonic generation of a millimeter-wave signal based on sextuple-frequency multiplication. AB - A millimeter-wave signal with sextuple-frequency multiplication of a microwave source is obtained with two cascaded optical modulators, which are driven by the same microwave source with phase deviation of pi/2 introduced by an electrical phase shifter. Without any optical filter, a wideband continuously tunable millimeter-wave signal is easily generated. PMID- 17410222 TI - Low-loss all-solid photonic bandgap fiber. AB - We report the fabrication and characterization of a new type all-solid photonic bandgap fiber. By introducing an index depressed layer around the high-index rod in the unit cell of photonic crystal cladding, transmission loss as low as 2 dB/km within the first bandgap is realized for the all-solid photonic bandgap fiber with a bandwidth of over 700 nm. The bend loss experiment shows that the photonic bandgap fiber is much less bend sensitive than single-mode fiber. PMID- 17410223 TI - Broadband regenerative wavelength conversion and multicasting using triple-stage semiconductor-based wavelength converters. AB - We demonstrate broadband wavelength conversion with a 320 nm operating wavelength range and channel spacing flexible wavelength-division-multiplexing (WDM) multicasting from a 1550 nm signal using a triple-stage cascaded semiconductor optical-amplifier-based wavelength converter. PMID- 17410224 TI - Optical low-coherence reflectometry for complete chromatic dispersion characterization of few-mode fibers. AB - A phase-sensitive optical low-coherence reflectometry (OLCR) technique is demonstrated to simultaneously measure the absolute chromatic dispersion values of each guided LP mode of a few-mode fiber. We show that the OLCR technique requires only short samples of fiber (<1 m) and has no need for high-ratio mode converters to reach an accurate wavelength-dependent group delay evolution of every mode. As an example we present for the first time to our knowledge a direct and complete analysis of few-mode fibers with high, low, positive, and negative modal dispersion values, leading to chromatic dispersion parameters in good agreement with theoretical predictions. PMID- 17410225 TI - Continuously spacing-tunable multiwavelength semiconductor-optical-amplifier based fiber ring laser incorporating a superimposed chirped fiber Bragg grating. AB - We investigate a flexibly tunable multiwavelength semiconductor-optical-amplifier based fiber ring laser with continuous wavelength spacing controllability incorporating a superimposed chirped fiber Bragg grating (CFBG). The wavelength spacing of a superimposed CFBG can be continuously controlled by symmetrically modifying the chirp bandwidth of the grating with the specially designed apparatus. We achieve a wide and continuous tuning range of the wavelength spacing from 0.35 to 0.78 nm. The continuous tunability of the wavelength spacing is measured to be ~ +/-0.033 nm/mm. By controlling the reflection bandwidth of the tunable CFBG, we can independently adjust the number of lasing channels from 2 to 23 at the wavelength spacing of 0.51 nm. PMID- 17410226 TI - In-fiber polarizer based on a long-period fiber grating written on photonic crystal fiber. AB - A novel in-fiber polarizer based on a long-period fiber grating (LPFG) is written by using a focused CO2 laser beam to notch a photonic crystal fiber periodically. Such a polarizer exhibits a high polarization extinction ratio of more than 20 dB over a wide wavelength range of ~11 nm near 1550 nm and a very low temperature sensitivity of 3.9 pm/ degrees C, which overcomes the disadvantages of the temperature sensitivity of other in-fiber polarizers created on conventional single-mode fiber. PMID- 17410227 TI - Ultraslow optical waveguiding in an atomic Bose-Einstein condensate. AB - We investigate waveguiding of ultraslow light pulses in an atomic Bose-Einstein condensate. We show that under the conditions of off-resonant electromagnetically induced transparency, waveguiding with a few ultraslow modes can be realized. The number of modes that can be supported by the condensate can be controlled by means of experimentally accessible parameters. Propagation constants and the mode conditions are determined analytically using a Wentzel-Kramers-Brillouin analysis. Mode profiles are found numerically. PMID- 17410228 TI - Mode-selective spatial filtering for increased robustness in a mode group diversity multiplexing link. AB - We propose a very simple optical method to reduce the cross talk among the channels of a mode group diversity multiplexing (MGDM) link. MGDM is an intensity modulation, direct detection, multiple-input, multiple-output technique that creates independent communication channels over a multimode fiber (MMF). The cross talk among the channels is mitigated electronically. However, by properly employing a lens between the output of a graded-index MMF and the detectors, we achieve mode-selective spatial filtering (MSSF) and optically reduce the cross talk. The robustness of the link is then increased when compared with an implementation without MSSF. This allows for a larger number of channels. PMID- 17410229 TI - Long-term femtosecond timing link stabilization using a single-crystal balanced cross correlator. AB - We demonstrate a self-aligned balanced cross correlator based on a single type-II phase-matched periodically poled KTiOPO4 crystal. The birefringence of the crystal generates a walk-off between the two orthogonally polarized pulses. This enables the balancing of the cross correlator with input pulses at the same center wavelength. As a first application of this single-crystal balanced cross correlator, we stabilized a 310 m long optical fiber link for timing distribution with long-term stable 10 fs precision. PMID- 17410230 TI - Passive combination of multiple beams in an optical fiber via stimulated Brillouin scattering. AB - We demonstrate passive combination of multiple beams in an optical fiber via stimulated Brillouin scattering (SBS). Four off-axis beams are combined in a long multimode optical fiber using a novel all-optical mount. The combined beam has the high spatial coherence properties of the LP(01) mode owing to the beam cleanup properties of SBS. The threshold for SBS when off-axis pumps are used is shown to be a factor of 13 times higher than for on-axis pump beams. We propose this method would improve the brightness of an array of fiber amplifiers. PMID- 17410231 TI - Orthogonal polarization shift keying label rewriting method in an all-optical label switching network. AB - A novel polarization shift keying label rewriting scheme in 43 Gbit/s all-optical label switching network by synchronous polarization modulation is proposed and experimentally demonstrated. The old label can be changed to a new one by just one single step without a label erasure process, which can simplify the network node structure. PMID- 17410232 TI - Generation of femtosecond pulses at 1350 nm by Cerenkov radiation in higher-order mode fiber. AB - We demonstrate a method of generating short pulses at 1350 nm by exciting Cerenkov radiation in a higher-order-mode fiber with a 1064 nm femtosecond fiber laser. We measure a 106 fs, 0.66 nJ output pulse. Cerenkov radiation in fibers allows for energy transfer between a soliton and a dispersive wave, providing an effective and engineerable platform to shift the wavelength of a femtosecond source. With appropriate design of the higher-order-mode fiber, this method of generating short pulses at 1350 nm can be extended to other wavelengths and to higher pulse energies. PMID- 17410233 TI - Development of a high-pulse-energy Q-switched Tm-doped double-clad fluoride fiber laser and its application to the pumping of mid-IR lasers. AB - A diode-pumped Q-switched Tm-doped double-clad fluoride fiber laser is reported providing up to 90 microJ pulse energy (160 ns, 100 kHz, i.e., 9 W of average power). The dependence of the fiber laser's repetition rate on pump power and modulator repetition rate was investigated. By amplification even higher pulse energies of 410 microJ could be generated. In a second stage of the setup the Q switched fiber laser serves as a pump for a gain-switched tunable Cr2+:ZnSe laser. The pulse energies reported are to the author's knowledge the highest generated by Tm-doped fluoride fiber lasers or amplifiers today. PMID- 17410235 TI - Spread-space spread-spectrum technique for secure multiplexing. AB - A novel technique for multiplexing complex images is proposed in which each image may be demultiplexed only if a set of random encryption keys is known. The technique utilizes the ability of the double random phase encoding method to spread a signals' energy in both the space and the spatial frequency domains in a controlled manner. To multiplex, images are independently encrypted with different phase keys and then superimposed by recording sequentially on the same material. Each image is extracted by using the particular key associated with it. During decryption the energy from the other images is further spread, making it possible to minimize its effects by using suitable filters. Wigner analysis is applied to the technique, and numerical results are presented. PMID- 17410236 TI - Determining parent radius and conic of an off-axis segment interferometrically with a spherical reference wave. AB - A simple interferometric method is presented for determining the parent radius of curvature (ROC) and conic constant (CC) of a conic surface. This method compares the test surface with a spherical reference wavefront having a ROC equal to the local sagittal, medial, or tangential ROC. The measured wavefront aberrations, particularly the astigmatism and coma, and local radii are used to determine the parent ROC and CC. This method does not require null optics or knowledge of the surface coordinate where the measurement is made. PMID- 17410237 TI - Programmable imaging with two-axis micromirrors. AB - We demonstrate a means of creating a digital image by using a two-axis tilt micromirror to scan a scene. For each different orientation we extract a single gray scale value from the mirror and combine them to form a single composite image. This allows one to choose the distribution of the samples, and so in principle a variable resolution image could be created. We demonstrate this ability to control resolution and projection by constructing a voltage table that compensates for the nonlinear response of the mirrors to the applied voltage. PMID- 17410238 TI - Electromagnetic analysis of cylindrical invisibility cloaks and the mirage effect. AB - We present a finite-element analysis of a diffraction problem involving a coated cylinder enabling the electromagnetic cloaking of a lossy object with sharp wedges located within its core. The coating consists of a heterogeneous anisotropic material deduced from a geometrical transformation as first proposed by Pendry [Science 312, 1780 (2006)]. We analyze the electromagnetic response of the cloak in the presence of an electric line source in p polarization and a loop of magnetic current in s polarization. We find that the electromagnetic field radiated by such a source located a fraction of a wavelength from the cloak is perturbed by less than 1%. When the source lies in the coating, it seems to radiate from a shifted location. PMID- 17410239 TI - Usage of turbulence for superresolved imaging. AB - We have used the turbulent flow of hot air to improve the diffraction limits of resolution in an imaging system. A time-multiplexing approach was applied and used the turbulent flow near the object to encode its spatially high frequencies. The turbulent perturbations were extracted and numerically implemented to decode the high-resolution image. Experimental verification demonstrated the proposed concept. PMID- 17410240 TI - Simple beam combination for stellar interferometry. AB - In stellar interferometry, image quality improves significantly with the inclusion of more telescopes and the use of phase closure. We demonstrate, using first coherent and then partially coherent white light, a compact and efficient pairwise combination of 12 or more beams. The input beams are lined up and spread through a cylindrical lens into a comb of parallel ellipses, which interferes with a perpendicular copy of itself to form a matrix of interferograms between all pairs. The diagonal elements show interference of each beam with itself, for intensity calibration. The measured white-light visibilities were high and stable. PMID- 17410241 TI - Facet braiding: a fundamental problem in integral imaging. AB - A rigorous explanation of a phenomenon that produces significant distortions in the three-dimensional images produced by integral imaging systems is provided. The phenomenon, which we refer to as the facet-braiding effect, has been recognized in some previous publications, but to our knowledge its nature has never been analyzed. We propose a technique for attenuating the facet-braiding effect. We have conducted experiments to illustrate the consequences of the facet braiding effect on three-dimensional integral images, and we show the usefulness of the proposed technique in eliminating this effect. PMID- 17410242 TI - Optical mosaic gratings made by consecutive, phase-interlocked, holographic exposures using diffraction from latent fringes. AB - We propose and demonstrate a method for fabricating large optical mosaic gratings, i.e., gratings made by consecutive, phase-interlocked, holographic exposures on single substrates. It takes advantage of the latent fringes generated by the preceding exposure to align the substrate for the following exposure. Good angular alignment accuracy and period consistency between the fringes recorded by consecutive exposures can be achieved by using the large-area moire pattern produced by the latent fringes and the alignment beams. Phase interlocking accuracy of less than 4 degrees between consecutive exposures was experimentally achieved by using the heterodyne detection technique. PMID- 17410243 TI - Similariton fiber laser with a hollow-core photonic bandgap fiber for dispersion control. AB - We report on an ytterbium-doped similariton fiber ring laser with a hollow-core photonic bandgap fiber for intracavity dispersion control. The oscillator is hybrid mode locked with a saturable Bragg reflector and by nonlinear polarization evolution. This scheme allows for an exact adjustment of the transmission characteristic to avoid the formation of bunched noiselike pulses while the self starting characteristic is preserved. The oscillator generates highly stretched similaritons at 1025 nm with a pulse energy above 1n J at a repetition rate of 21.9 MHz. PMID- 17410244 TI - 10 J energy-level optically pumped XeF(C-A) laser with repetition mode. AB - An optically pumped XeF(C-A) laser with 0.1-1 Hz pulse-repetition mode is demonstrated. The maximum output energy is up to 18.7 J, which corresponds to the conversion efficiency of the stored electrical energy to laser output energy of 0.25%. The laser active medium is optically excited by vacuum-ultraviolet radiation of two single-channel surface discharges initiated along opposite walls of the laser cell. The laser resonator output coupling is 10%, and the initial XeF2 concentration in the active gas mixture is ~1 x 10(17) cm(-3). The duration of the observed laser pulses is 0.8 to 1.2 micros, and the spectral band is 470 500 nm. PMID- 17410245 TI - Dual-frequency single-axis laser using a lead lanthanum zirconate tantalate (PLZT) birefringent etalon for millimeter wave generation: beyond the standard limit of tunability. AB - We demonstrate the generation of optically carried, broadly tunable, millimeter wave signals with a dual-frequency single-axis Nd:YAG laser. A frequency difference as high as 127 GHz is reached thanks to an intracavity electro optically tunable etalon made of lead zirconate tantalate (PLZT) ceramic. We show that the available frequency range is actually limited by the bandwidth of the amplification medium, namely, far beyond the usually accepted free spectral range value in the case of a single-axis laser. Both coarse discrete and fine continuous tunabilities are obtained with the same voltage-controlled device, opening the way to widely tunable low-phase-noise optically carried submillimeter or even terahertz sources. PMID- 17410246 TI - High-Q-preserving coupling between a spiral and a semicircle micro-cavity. AB - We present an efficient design for direct coupling between a spiral-shaped and a semicircle-shaped microcavity (micro-cavity) as an alternative to traditional evanescent wave coupling for planar integrated photonic technology. We observe the preservation of the high Q-value of the spiral oscillator when coupled to a semicircle under current injection using an AlGaAs single-quantum-well heterostructure. With slight alterations to the directly coupled micro-cavity configuration, such as coupling shape and overlap distance, the number of observed modes and output intensity are changed. AlGaAs and InGaN spiral-shaped microcavities have unidirectional emission normal to the spiral notch. PMID- 17410247 TI - Short pulse eye-safe laser with a stimulated Raman scattering self-conversion based on a Nd:KGW crystal. AB - We present the results of a solid self-Raman laser based on a Nd:KGW crystal that is transversely pumped by laser diode bars. A beam of an eye-safe laser with a 31.8 mJ output energy and a 2.0 ns pulse width was obtained by applying a special s-polarized reflective resonator configuration in which the length of the Raman resonator was shorter than that of the fundamental radiation resonator. The eye safe laser has the highest output energy and the shortest pulse width among the Nd:KGW lasers ever reported. PMID- 17410248 TI - Beat-note jitter suppression in a dual-frequency laser using optical feedback. AB - An efficient locking technique based on optical feedback is demonstrated to suppress jitter on the rf beat note between the two modes of a dual-frequency Yb:Er glass laser. The method consists of a self-injection process in which one selected mode serves as a master oscillator to lock and stabilize the second mode via a frequency-shifted optical feedback. The beat note adjusted near 170 MHz was stabilized with an accuracy of 250 mHz using an optical feedback loop with a double pass through an acousto-optic modulator. The beating note can be tuned over 300 kHz by controlling the reference oscillator. The extensions and limitations of the technique are discussed. PMID- 17410249 TI - Large area confocal microscopy. AB - Imaging large tissue areas with microscopic resolution in vivo may offer an alternative to random excisional biopsy. We present an approach for performing confocal imaging of large tissue surface areas using spectrally encoded confocal microscopy (SECM). We demonstrate a single-optical-fiber SECM apparatus, designed for imaging luminal organs, that is capable of imaging with a transverse resolution of 2.1 microm over a subsurface area of 16 cm2 in less than 1 min. Due to the unique probe configuration and scanning geometry, the speed and resolution of this new imaging technology are sufficient for comprehensively imaging large tissues areas at a microscopic scale in times that are appropriate for clinical use. PMID- 17410250 TI - Simple and versatile dual-signal wave optical parametric oscillator. AB - A synchronously pumped optical parametric oscillator that is able to deliver a signal pulse at two different wavelengths either simultaneously or even one at a time is presented. As the operation exploits the group-velocity mismatch between the interacting signal and pump pulses, just a cavity-length adjustment is required to switch between the three regimes. The widely tunable signal pulses are shown to be synchronous and near transform limited. PMID- 17410251 TI - Low-voltage tunable second-harmonic generation in an x-cut periodically poled lithium niobate waveguide. AB - We report a new method of tuning the second-harmonic signal generated in a periodically poled lithium niobate (PPLN) waveguide. This technique relies on the recent progress in the fabrication of PPLNs on x-cut substrates along with periodical coplanar electrodes to reduce the tuning voltage while preserving the conversion efficiency. Our scheme exploits a type I interaction implemented in a titanium waveguide to impede cancellation of the electro-optical coefficients and to minimize the group-velocity mismatch between the first- and second-harmonic modes. The tuning range covers 58 nm with an applied voltage of +/-150 V. PMID- 17410252 TI - Four-wave parametric oscillation in sodium vapor by electromagnetically induced diffraction. AB - We have observed a novel type of parametric oscillation in sodium atomic vapor where four off-axis signal waves simultaneously build up under resonant and counterpropagating pump beams with elliptical beam profiles. The four waves, two of them Stokes shifted and the other two anti-Stokes shifted, have similar output powers of up to 10 mW with a conversion efficiency of 30% and are parametrically coupled by electromagnetically induced diffraction. PMID- 17410253 TI - All-solid-state 704 mW continuous-wave yellow source based on an intracavity, frequency-doubled crystalline Raman laser. AB - Continuous-wave operation at 588 nm of a diode-pumped Nd:GdVO4 laser with intracavity Raman shifting [in KGd(WO4)2, KGW] and frequency-doubling (in LiB3O5, LBO) is reported. A maximum cw power at 588 nm of 704 mW was obtained for diode pump powers of 13.7 W. Quasi-cw yellow powers up to 1.57 W at a 50% duty cycle (to reduce thermal load in the laser crystal) indicate that power scaling to over 1 W is feasible. PMID- 17410254 TI - Nonlinear dynamics of negatively refracted light in a resonantly absorbing Bragg reflector. AB - The evolution of nonlinear light fields traveling inside a resonantly absorbing Bragg reflector is studied by use of Maxwell-Bloch equations. Numerical results show that a pulse initially resembling a light bullet may effectively experience negative refraction and anomalous dispersion in the resonantly absorbing Bragg reflector. PMID- 17410255 TI - Observation of interference between four-wave mixing and six-wave mixing. AB - We report generation of four-wave mixing enhanced by electromagnetically induced transparency and optical pumping in a ladder-type atomic system. When two pumping laser beams are used to form a conjugate small-angle static grating, both four wave and six-wave mixing processes are shown to exist at the same time. Interference between these two nonlinear wave-mixing signals is experimentally demonstrated. PMID- 17410256 TI - Cavity-dumped intracavity-frequency-doubled Yb:YAG thin disk laser with 100 W average power. AB - We report on a cavity-dumped Yb:YAG thin disk laser with intracavity-frequency doubling to provide pulses in the millijoule energy range at a repetition rate of up to 100 kHz. The maximum average output power at 515 nm was 102 W with pulses of a pulse length of 300 ns. An additional advantage of the presented laser setup is the wide tunability of the pulse duration. PMID- 17410257 TI - Helmholtz solitons at nonlinear interfaces. AB - Reflection and refraction of spatial solitons at dielectric interfaces, accommodating arbitrarily angles of incidence, is studied. Analysis is based on Helmholtz soliton theory, which eliminates the angular restriction associated with the paraxial approximation. A novel generalization of Snell's law is discovered that is valid for collimated light beams and the entire angular domain. Our new theoretical predictions are shown to be in excellent agreement with full numerical simulations. New qualitative features of soliton refraction and limitations of previous paraxial analyses are highlighted. PMID- 17410258 TI - Widely tunable 3.4 microm band difference frequency generation using apodized chi(2) grating. AB - We achieved apodization in a quasi-phase-matched wavelength converter. The new design yields a large bandwidth and a flat phase-matching response with a high conversion efficiency. Using the method, we demonstrate widely tunable 3.4 microm band difference frequency generation in a LiNbO3 ridge waveguide. PMID- 17410259 TI - Ghost imaging with intense fields from chaotically seeded parametric downconversion. AB - We present the first experimental demonstration, to the best of our knowledge, of ghost imaging realized with intense beams generated by a parametric downconversion interaction seeded with pseudothermal light. As expected, the real image of the object is reconstructed satisfying the thin-lens equation. We show that the experimental visibility of the reconstructed image is in accordance with the theoretically expected one. PMID- 17410260 TI - Experimental demonstration of mode structure in ultralong Raman fiber lasers. AB - We present the first experimental demonstration of a resolvable mode structure with spacing c/2nL in the RF spectra of ultralong Raman fiber lasers. The longest ever demonstrated laser cavity (L=84 km), RF peaks of ~100 Hz width and spacing ~1 kHz have been observed at low intracavity powers. The width of the peaks increases linearly with growing intracavity power and is almost independent of fiber length. PMID- 17410261 TI - Mid-infrared difference-frequency generation of ultrashort pulses tunable between 3.2 and 4.8 microm from a compact fiber source. AB - We report single-pass difference-frequency generation of mid-infrared femtosecond pulses tunable in the 3.2-4.8 microm range from a two-branch mode-locked erbium doped fiber source. Average power levels of up to 1.1 mW at a repetition rate of 82 MHz are obtained in the mid infrared. This is achieved via nonlinear mixing of 170 mW, 65 fs pump pulses at a fixed wavelength of 1.58 microm, with 11.5 mW, 40 fs pulses tunable in the near-infrared range between 1.05 and 1.18 microm. These values indicate that the tunable near-infrared input component is downconverted with a quantum efficiency that exceeds 30%. PMID- 17410262 TI - Imprinted diffractive optics for terahertz radiation. AB - Terahertz diffractive optic elements have been fabricated in polypropylene by imprinting with a silicon master. A silicon master is created with eight phase levels for high diffraction efficiency and etched using inductively coupled plasma reactive ion etching. This technique enables the rapid replication of complex optical structures in a high transmission material. Excellent replication of multilevel high efficiency Fresnel lenses is shown. The resulting lenses were tested with a 2 THz quantum cascade laser. The signal strength at the focus was 70 times the base signal strength. PMID- 17410263 TI - Fractionation of polydisperse colloid with acousto-optically generated potential energy landscapes. AB - The motion of colloidal particles on a periodic optical potential energy landscape in the presence of an external driving force may result in particle separation. In contrast to recent methods of holographic or interferometric generation of such landscapes, we use an acousto-optic deflector to create two dimensional landscapes. We present what is believed to be the first experimental realization of fractionation with simultaneous sorting of four different sizes of colloidal microparticle into laterally separated parallel laminar streams. PMID- 17410264 TI - Coupled whispering gallery modes in a multilayer-coated microsphere. AB - Whispering gallery modes in a microsphere coated with three layers of high, low, and high refractive indices (RIs) are considered. Coastal modes and inland modes, centered on the outer and inner high-RI layers, exist as different radial modes. At right values of RI and thickness of the three layers, an inland mode and a coastal mode couple to produce a radial distribution of the photonic field that resembles bonding and antibonding orbitals. The coupling occurs across a layer of the middle low-RI layer, much thicker than the wavelength of light. The coupling is analyzed in a quantum-mechanical analog of a one-dimensional particle in a double-well potential. PMID- 17410265 TI - Transient response of blue organic electroluminescence devices with short fluorescence lifetime of substituted phenyl/vinyl compound as an emissive layer. AB - We have demonstrated a short fluorescence lifetime of the substituted phenyl/vinyl compound, 1,4-bis[2-[4-[N,N-di(p-tolyl)amino]phenyl]vinyl]benzene (DSB). The fluorescence lifetime of the 0.5 mol.% DSB-doped 4,4'-bis(9 carbazolyl)biphenyl film is 1.2 ns, which is desirable for organic light-emitting diode (OLED) light sources for optical interconnect applications. We have also examined frequency dependences on the electroluminescence (EL) intensity of the OLED and the photoluminescence (PL) intensity of the DSB film. The -3 dB cutoff frequency of the EL intensity is about 3 MHz for the optimized device, and the 3dB cutoff frequency of the PL intensity is about 160 MHz for the optically pumping DSB film. PMID- 17410266 TI - Edge-emitting photonic crystal double-heterostructure nanocavity lasers with InAs quantum dot active material. AB - We report what is, to the best of our knowledge, the first demonstration of an edge-emitting photonic crystal nanocavity laser that is integrated with a photonic crystal waveguide. This demonstration is achieved with a double heterostructure photonic crystal nanocavity incorporating an InAs quantum dot active region. PMID- 17410267 TI - Enhanced optical forces between coupled resonant metal nanoparticles. AB - We investigate numerically the optical forces between noble metal nanoparticles sustaining localized surface plasmon resonances. Our results first point out enhanced binding optical forces compared with dielectric nanoparticles and nonresonant metallic nanoparticles. We also show that under suitable illumination conditions, short-range forces tend to make the nanoparticles cluster, leading to intense and localized hot spots in the interstices. This effect corroborates recent experimental observations of an enhanced Raman signal in trapped metal sphere ensembles. PMID- 17410268 TI - Radiation pattern of cylindrically symmetric scalar Laguerre-Gauss beams. AB - The exact full-wave generalization of the cylindrically symmetric scalar real argument Laguerre-Gauss beam is determined. The radiation intensity of the resulting Laguerre-Gauss wave is deduced, and the main characteristics of the radiation intensity pattern are described. The total power P(n) is evaluated, where n is the radial mode number. By the use of 1/P(n) as the quality parameter, the characteristics of the quality of the paraxial beam approximation to the full Laguerre-Gauss wave are discussed. PMID- 17410269 TI - Electromagnetically induced self-imaging. AB - We study the self-imaging and image-transforming properties of a probe field in a cold atomic medium with electromagnetically induced transparency (EIT). Due to the similarities between the gradient-index medium and the inhomogeneous index distribution of an EIT medium under the conditions of a negative probe detuning and a Gaussian control field, we find based on analytical investigations that there exists a kind of electromagnetically induced self-imaging phenomenon in cold atomic media. Numerical simulations clearly show that electromagnetically induced self-imaging is observable and controllable. PMID- 17410270 TI - Differential-phase quantum key distribution experiment using a series of quantum entangled photon pairs. AB - We report what we believe to be the first differential-phase quantum key distribution experiment using a series of quantum entangled photon pairs. We employed two outstanding techniques. As an entangled photon source, we used a 1.5 microm band entangled photon pair source based on spontaneous four-wave mixing in a cooled dispersion-shifted fiber. As receivers, photon pairs were actively phase modulated with LiNbO3 phase modulators followed by very stable planar light-wave circuit Mach-Zehnder interferometers, which provided two nonorthogonal measurements. As a consequence, we successfully generated sifted keys with a quantum bit error rate of 8.3% and a key generation rate of 0.3 bit/s and revealed the feasibility of this QKD scheme. PMID- 17410271 TI - Optimized matrix inversion technique for the T-matrix method. AB - We suggest a new approach to calculate the inverse matrix in scattering calculations using the T-matrix method. Instead of inversion of the full matrix, we suggest the inversion of two matrices, each of which contains half the number of rows. This approach allows significant time savings and a noticeable increase of the precision of scattering calculations due to fewer arithmetical operations. An iterative method can be applied to matrices whose dimension is also divisible by factors of 2, which can further increase the time savings and accuracy. PMID- 17410272 TI - Accurate determination of microparticle size using Fourier transform of light scattering spectrum over wavenumber. AB - We discovered that the far-field light scattering spectra of microparticles over wavenumber at a certain angle could be decomposed into periodic components, with an oscillation frequency linearly dependent on the particle size at each angle. Based on this observation, we propose and experimentally demonstrate a new Fourier transform technique for microparticle size determination. This technique is simple, fast, robust in its data processing algorithm, and flexible in its detection system. PMID- 17410273 TI - Upconversion luminescence of CdTe nanocrystals by use of near-infrared femtosecond laser excitation. AB - We study the steady-state and time-resolved luminescent properties of CdTe nanocrystals by one- and two-photon excitation with a femtosecond laser. We observe that 1208 nm excitation causes a shift of the emission peak of about 20 nm to the infrared compared with 400 nm laser excitation. It is found that upconversion luminescence is composed of a photoinduced trapping and a band edge excitonic state and produces the observation of biexponential decay kinetics. We conclude that the redshift of the emission peak is caused by the relative change in luminescence intensity between excitonic and trapping states. PMID- 17410274 TI - External cavity quantum cascade laser for quartz tuning fork photoacoustic spectroscopy of broad absorption features. AB - We demonstrate mid-infrared spectroscopy of large molecules with broad absorption features using a tunable external cavity quantum cascade laser. Absorption spectra for two different Freons are measured over the range 1130-1185 cm(-1) with 0.2 cm(-1) resolution via laser photoacoustic spectroscopy with quartz tuning forks as acoustic transducers. The measured spectra are in excellent agreement with published reference absorption spectra. PMID- 17410275 TI - Pulse broadening of the femtosecond pulses in a Gaussian beam passing an angular disperser. AB - A general analytical formula has been found to describe the evolution of the pulse width of the femtosecond pulses in a Gaussian beam after passing an angular disperser without the assumption of well collimation. This formula is experimentally verified by measuring the pulse width with an autocorrelator based on two-photon absorption. It is found that the effect of the spectral lateral walk-off and group delay dispersion on the pulse-width evolution, and its dependence on the distance traveled, are substantially different when the beam has not been well collimated than from when it has been collimated. These differences result from the decaying nature of the angular dispersion of the Gaussian beam sent across a distance. PMID- 17410276 TI - Dispersion control over the ultraviolet-visible-near-infrared spectral range with HfO2/SiO2-chirped dielectric multilayers. AB - We report the first realization, to the best of our knowledge, of a chirped multilayer dielectric mirror providing dispersion control over the spectral range of 300-900 nm and the first use of hafnium oxide in a chirped mirror. The technology opens the door to the reliable and reproducible generation of monocycle laser pulses in the blue-violet spectral range, will benefit the development of optical waveform and frequency-comb synthesizers over the ultraviolet-visible-near-infrared spectral range, and permits the development of ultrabroadband-chirped multilayers for high-power applications. PMID- 17410277 TI - Single attosecond pulse generation from asymmetric molecules with a multicycle laser pulse. AB - A new scheme for single attosecond pulse generation from asymmetric molecules with a multicycle laser pulse is proposed. It is shown that both even and odd harmonics are generated from the asymmetric molecules, and the attosecond pulses are produced every full cycle of the driving laser field rather than each half cycle. By filtering the harmonics in the cutoff of the spectrum, a single attosecond pulse can be obtained with a multicycle laser pulse with a duration of 2 times longer than the few-cycle pulse conventionally used. PMID- 17410278 TI - Using correlational analyses to improve prevention strategies based on survey data from youth. AB - Community coalition prevention models often select interventions based on the types of risk factors (outcome predictors) that are elevated. Variances and correlations between predictors and targeted behaviors also may vary between communities and provide information to improve the selection of interventions. Community differences in risk factor levels and correlations between predictors and problem behaviors were examined using a child self-report computer assessment (ALEXSA); prevention.psu.edu/people/ALEXSA.htm). Three school-based subsamples of children were studied. Means, prevalences, and correlations differed significantly between samples. Discussion addresses developmental considerations and illustrates how correlations between predictors and problem behaviors might improve the selection of interventions. This study is preliminary and should be replicated with larger community samples, more indicated/selected sample, and in more communities. PMID- 17410279 TI - Plasma C-reactive protein in nonobese children with obstructive sleep apnea before and after adenotonsillectomy. AB - STUDY OBJECTIVE: Sleep-disordered breathing (SDB) is a prevalent condition in children and is associated with increased cardiovascular morbidity. Circulating levels of C-reactive protein (CRP), a proinflammatory protein, are associated with increased risk for atherosclerosis. Plasma CRP levels in snoring children have yielded conflicting results, such that it remains unclear whether OSA is mechanistically involved in such elevations of CRP. METHODS: Consecutive nonobese children with polysomnographically demonstrated obstructive sleep apnea underwent blood draws in the morning after their corresponding sleep studies on 2 occasions, namely at diagnosis of obstructive sleep apnea and 10 to 14 weeks after adenotonsillectomy. High-sensitivity CRP serum concentrations were determined within 2 to 3 hours after collection, using a particle-enhanced turbidimetric immunoassay technique. RESULTS: Twenty children with obstructive sleep apnea (mean age 7.3 +/- 1.9 years; 55% boys; relative body mass index: 88% +/- 12.0%) with a mean apnea-hypopnea index at diagnosis of 15.6 +/- 2.9 events per hour of total sleep time and nadir SaO2 of 82.3% +/- 2.5% were included. Mean initial CRP levels at obstructive sleep apnea diagnosis were 0.67 +/- 0.21 mg/dL and decreased to 0.23 +/- 0.07 mg/dL after adenotonsillectomy (p < .05), along with significant decreases in measured apnea-hypopnea index (2.2 +/- 0.8 events/h of total sleep time; p < .01) and improved oxygenation (mean nadir SaO2 values: 88.6% +/- 1.9%; p < .01). CONCLUSIONS: Obstructive sleep apnea is frequently associated with increases in CRP levels that are reversible upon treatment. Thus, obstructive sleep apnea induces a systemic inflammatory response in children, which, if left untreated, may potentially lead to cardiovascular morbidity. PMID- 17410280 TI - Treating agrammatic aphasia within a linguistic framework: Treatment of Underlying Forms. AB - BACKGROUND: Formal linguistic properties of sentences-both lexical, i.e., argument structure, and syntactic, i.e., movement-as well as what is known about normal and disordered sentence processing and production, were considered in the development of Treatment of Underlying Forms (TUF), a linguistic approach to treatment of sentence deficits in patients with agrammatic aphasia. TUF is focused on complex, non-canonical sentence structures and operates on the premise that training underlying, abstract, properties of language will allow for effective generalisation to untrained structures that share similar linguistic properties, particularly those of lesser complexity. AIMS: In this paper we summarise a series of studies focused on examining the effects of TUF. METHODS #ENTITYSTARTX00026;PROCEDURES: In each study, sentences selected for treatment and for generalisation analysis were controlled for their lexical and syntactic properties, with some structures related and others unrelated along theoretical lines. We use single-subject experimental designs-i.e., multiple baseline designs across participants and behaviours-to chart improvement in comprehension and production of both trained and untrained structures. One structure was trained at a time, while untrained sentences were tested for generalisation. Participants included individuals with mild to moderately severe agrammatic, Broca's aphasia with characteristic deficits patterns. OUTCOMES #ENTITYSTARTX00026; RESULTS: Results of this work have shown that treatment improves the sentence types entered into treatment, that generalisation occurs to sentences which are linguistically related to those trained, and that treatment results in changes in spontaneous discourse in most patients. Further, we have found that generalisation is enhanced when the direction of treatment is from more to less complex structures, a finding that led to the Complexity Account of Treatment Efficacy (CATE, Thompson, Shapiro, Kiran, & Sobecks, 2003). Finally, results of recent work showing that treatment appears to affect processing of trained sentences in real time and that treatment gains can be mapped onto the brain using functional magnetic resonance imaging (fMRI) are discussed. CONCLUSIONS: These findings indicate that TUF is effective for treating sentence comprehension and production in patients who present with language deficit patterns like those seen in our patients. Patients receiving this treatment show strong generalisation effects to untrained language material. Given the current healthcare climate, which limits the amount of treatment that aphasic patients receive following stroke, it is important that clinicians deliver treatment that results in optimal generalisation in the least amount of time possible. PMID- 17410281 TI - HIV-RISK BEHAVIORS AMONG PSYCHIATRICALLY HOSPITALIZED ADOLESCENTS WITH AND WITHOUT COMORBID SUD. AB - OBJECTIVE: The purpose of this study was to examine HIV-risk behaviors among a sample of psychiatric inpatient adolescents with and without comorbid SUD. METHOD: 239 adolescents (60.7% female; mean age=15.3) were interviewed while hospitalized in a psychiatric inpatient treatment facility. Adolescents with and without substance use disorder (SUD) were compared on a number of HIV-risk behaviors and the association between HIV-risk behaviors and other types of mental health problems were examined. RESULTS: Compared to those without SUD, adolescents with SUD, after controlling for age, gender, and other psychiatric disorders, were at an increased risk for being currently sexually active (AOR=2.01, CI=1.00-4.04, p<.05) and for having used alcohol or drugs prior to last sexual intercourse (AOR=5.48, CI=1.91-15.72, p<.01). HIV-risk behaviors were more evident among adolescents with externalizing disorders and those with higher levels of self-reported distress. CONCULSION: Adolescents in psychiatric settings, especially those with substance use disorders and/or externalizing problems, are an important population for whom prevention efforts are needed to reduce the risk of HIV infection. PMID- 17410282 TI - Noninvasive detection through REMS-PCR technique of K-ras mutations in stool DNA of patients with colorectal cancer. AB - BACKGROUND AND AIMS: Tumor exfoliated cells that shed into stool are attractive targets for molecular screening and early detection of colon malignancies. Many studies have suggested that the detection of activated ras may have diagnostic or prognostic importance. The aim of this study was to establish the suitability for use in diagnostic laboratories of the noninvasive screening test of K-ras mutation determination in the stool and its routine prognostic value in colorectal cancer. METHODS: Paired stool and tissue specimens obtained after polypectomy and colorectal biopsy from 28 patients diagnosed solely by histopathological findings with primary colorectal carcinoma, were prospectively studied for K-ras codon 12 mutations by restriction endonuclease-mediated selective (REMS)-PCR. RESULTS: DNA was obtained in 28 of tissue samples (100%) and 26 of stool samples (92.8%). K-ras codon 12 mutation was seen in 14 (50.0%) paired stool and tissue samples. Mutation detection was possible in 1000-fold excess of wild-type sequence. These results may be important in the design of genetic screening programs, determination of prognosis, early detection and treatment for patients with colon malignancy. CONCLUSIONS: The sensitivity and specificity of K-ras determination on stool-derived DNA in patients with colorectal carcinoma, support the opportunity of a large-scale trial to validate its use as a screening test. REMS- PCR is not labor intensive, but a sensitive, rapid, and robust assay for the detection of point mutations, and was introduced by us in a routine diagnostic laboratory. PMID- 17410283 TI - Clinical significance of p53, K-ras and DCC gene alterations in the stage I-II colorectal cancers. AB - BACKGROUND AND AIMS: Genetic alterations of p53, K-ras and DCC genes have a pivotal role in the colorectal cancer progression. The aim of this study was to clarify the association between K-ras mutations, p53 aberrations and DCC loss of heterozygosity (LOH), with the patient outcome and tumor characteristics in 43 stage I-II colorectal cancer patients. METHODS: Mutations in exons 5-8 of the p53 gene and codon 12 and/or 13 of the K-ras gene were assayed by PCR-SSCP and then confirmed by DNA sequencing. DCC LOH was studied by PCR-RFLP, while p53 immunohistochemistry was also made. RESULTS: Mutations of the p53 gene were found in 14 (32.5%) tumors. Five (12%) cases showed mutation of the K-ras gene. Nuclear staining of p53 was found in 22 (51 %) cases. DCC LOH was found in 5 (12%) cases. Cases with guanine to thymine substitution that occurred in K-ras codon 12 and DCC LOH were found to be more aggressive than other cases with codon 12 mutations or DCC wild-type phenotype. Many tumors with p53 over-expression were localized on the left side of the colon (p=0.005). The stage of the tumor was higher in patients who died during the follow-up period, when compared to the ones who have survived. CONCLUSIONS: Although none of these genetic alterations showed a significant prognostic value, specific mutation of K-ras gene and DCC LOH phenotype might have a predictive prognostic implication in colorectal cancer. Furthermore, different etiopathogenetic mechanisms might be involved in the tumorigenesis of the left and right colon. PMID- 17410284 TI - Long term follow-up of a large cohort of inactive HBsAg (+)/ HBeAg (-)/ anti-HBe (+) carriers in Greece. AB - AIM: To investigate the long-term outcome and the risk of progression to chronic hepatitis B in inactive hepatitis B surface antigen carriers. MATERIAL AND METHODS: A total of 307 HBsAg (+)/HBeAg (-)/antiHBe (+) subjects with initially normal alanine aminotransferase (ALT) levels and undetectable/ low serum HBVDNA with hybridization assay and later with PCR (10(5) copies/ml), were followed-up every 6 months for a period of 3 to 21 years (7.45 +/- 3.75 years). RESULTS: 234 out of the 307 HBsAg (+) patients (76.2%) had persistently normal ALT and undetectable / low (10(5) copies/ml) HBVDNA during follow-up. In 73 patients (23.8%), a reactivation of the disease with elevated ALT and positive HBVDNA (> (10(5)copies/ml) was recorded during the follow up. Thirty-five out of 73 patients underwent liver biopsy, while 22 of them received treatment. Twenty-four patients (7.8%) lost HBsAg after a mean of 7.4 +/- 3.6 years. Regarding the complications of chronic hepatitis B, only one patient developed compensated cirrhosis and no one developed HCC. CONCLUSIONS: Our results show that in almost 24% of inactive chronic hepatitis B carriers reactivation of the disease may occur even after many years. However the risk of liver-related complications is very low in these subjects. PMID- 17410285 TI - Efficacy, tolerability and predictive factors for early and sustained virologic response in patients treated with weight-based dosing regimen of PegIFN alpha-2b ribavirin in real-life healthcare setting. AB - BACKGROUND AND AIM: Increasing evidence to date highlights that individualized treatment regimens with pegylated interferon (PegIFN) and ribavirin represent a better approach for patients nowadays showing negative predictive factors for sustained virological response. The aims of this study were to assess the rate of early (EVR) and sustained virological response (SVR), tolerability and baseline predictive factors associated with EVR and SVR in patients with chronic hepatitis C treated with individualized weight-based dosing regimen for both PegIFN alpha 2b and ribavirin. METHODS: The observational analysis included 234 consecutive patients with chronic hepatitis C genotype 1 treated with PegIFN alpha-2b and ribavirin on an out-patient basis between January 2003-March 2006. RESULTS: The mean age of the study group was 49.5 years, and 35% were male patients; the group was slightly overweight (mean BMI=26.5 kg/sq.m). EVR was achieved in 84.6% (198/234 patients). The end-of-treatment and sustained biochemical responses were 76.3% and 66.1%, respectively. At the end of follow-up, an overall intent-to treat SVR was achieved by 71 of 127 patients (in 55.9%). Lower baseline (< 1,000 000 IU/mL) HCV viral load was the only predictive factor associated with EVR (p=0.04); absent or mild fibrosis (F0-1) and a low histological activity (HAI < 8) were independently associated with SVR. Side effects resulted in PegIFN and ribavirin dose reductions in 9.4% and, respectively, 18.1%, but definitive discontinuation of therapy was necessary only in 8.7% of patients. CONCLUSION: PegIFN alpha-2b and ribavirin can be safe and successful when using a weight based dosing regimen, leading to high response rates even in overweight patients. PMID- 17410286 TI - The FibroTest value in discriminating between insignificant and significant fibrosis in chronic hepatitis C patients. The Romanian experience. AB - AIM: To assess the diagnostic value of FibroTest to discriminate between insignificant and significant fibrosis in order to avoid the liver biopsy currently used for selection of chronic hepatitis C patients eligible for antiviral therapy. PATIENTS AND METHODS: A retrospective study was carried out in 206 chronic hepatitis C patients with liver biopsy performed before starting antiviral therapy and concomitant serum stored at -80 degrees C. Liver fibrosis was evaluated according to the METAVIR scoring system on a scale of F0 to F4. Biochemical markers assessed were: alpha 2 macroglobulin (alpha 2-MG), apolipoprotein A1 (Apo-A1), haptoglobin (Hapto), gamma-glutamyltransferase (GGT), total bilirubin (TB). The FibroTest score was computed after adjusting for age and gender. Predictive values and ROC curves were used to assess the accuracy of FibroTest results. RESULTS: Alpha 2-MG, apo-A1, Hapto and gender were independent predictors for significant fibrosis. For FibroTest the observed area under ROC (ObAUROC) for the discrimination between minimal or no fibrosis (F0-F1) and significant fibrosis (F2-F4) was 0.782 (+/- 95 CI: 0.716-0.847) for a cutoff value 0.47. The sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV) of the FibroTest to differentiate significant from insignificant fibrosis were 80.2; 63.2; 78.9 and 65.8, respectively. The adjusted AUROC (AdAUROC) according to the prevalence of each individual stage of fibrosis was 0.856. CONCLUSION: FibroTest could be an alternative to biopsy in most patients with chronic hepatitis C. It requires a strict adherence and observance of the technical recommendations for the assays of biochemical markers in order to avoid analytical variability. PMID- 17410287 TI - Therapeutic options in non-alcoholic steatohepatitis (NASH). Are all agents alike? Results of a preliminary study. AB - AIM: The evaluation of the efficacy of ursodeoxycholic acid (UDCA), pentoxifylline, losartan, and atorvastatin in non-alcoholic steatohepatitis (NASH) treatment. METHOD: 48 patients (25 males/23 females, aged 55 +/- 7.54 years) with histologically confirmed NASH were enrolled between 2001 and 2005. The batch was divided into four groups: A (10 dyslipidemic patients, receiving atorvastatin 10 mg/day), P (13 nonhypertensive/ nondyslipidemic patients receiving pentoxifylline 400 mg bid), L (12 hypertensive patients, treated with losartan, 50 mg/day) and U (13 nonhypertensive patients receiving UDCA 15 mg/kg/day). Mean duration of treatment was 37.8 +/- 5.4 weeks. Body mass index, liver biopsy and serum level of alanine-aminotransferase (ALT), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (AP), total cholesterol (TC) and triglycerides (TG) were determined at inclusion and at the end of treatment. Liver biopsy samples were evaluated for necroinflammation, steatosis and fibrosis (Brunt's score). RESULTS: In group A, a significant reduction of ALT, GGT, TC and AP was noticed. Histology showed diminished steatosis, but no improvement of fibrosis and necroinflammation. In groups P and L we found a reduction of mean ALT and GGT levels and necroinflammatory score. Group U presented a significant reduction in ALT and GGT levels, without improvement in steatosis, necroinflammation or fibrosis. CONCLUSION: Atorvastatin and losartan proved to be efficient in the treatment of dyslipidemia- and hypertension-associated NASH, by improving both biochemical parameters and steatosis/ necroinflammation. Pentoxifylline showed similar efficacy in non-hypertensive/non-dyslipidemic patients, while UDCA did not improve the histological score although it improved the biochemical parameters. PMID- 17410288 TI - Glycogen storage disease type I--between chronic ambulatory follow-up and pediatric emergency. AB - BACKGROUND AND AIMS: To describe the characteristics of patients with type I glycogenosis, the presentation types, the main clinical signs, the diagnostic criteria and also the disease outcomes on long term follow-up. METHODS: The study group consisted of 6 patients (medium age 3 years 6 months) admitted in hospital between 2001 and 2005 and followed-up for 1 to 5 years. The sex ratio was 1:1. RESULTS: The referral reasons varied from hepatomegaly incidentally discovered (3 of 6 patients) to abdominal pain (4 of 6 patients), growth failure (3 of 6 patients), symptoms of hypoglycemia (3 of 6 patients), recurrent epistaxis (1 patient). Hepatomegaly was present in all cases. Biological profile: hypoglycemia, increased transaminase values, hypertriglyceridemia, lactic acidosis, normal uric acid levels. Two patients had neutropenia and other two had increased glomerular filtration rate. Liver biopsy showed glycogen-laden hepatocytes and markedly increased fat. Four patients had type Ia and 2 patients type Ib glycogenosis. The therapy consisted of: diet, ursodeoxycholic acid, granulocyte colony-stimulating factor, broad spectrum antibiotics for those with type Ib glycogenosis. The follow-up parameters were clinical, biological, imaging. Metabolic interventions and antiinfectious therapy were necessary. All patients are alive, two of them on the waiting list for liver transplantation. CONCLUSIONS: Glycogen storage disease type I is a rare condition, but with possible life-threatening consequences. It has to be kept in mind whenever important hepatomegaly and/or hypoglycemia are present. PMID- 17410289 TI - Preemption dimensional study for obtaining statistically significant results for the variation of gamma-glutamyl-transferase during ovarian stimulation. AB - BACKGROUND: Ovarian stimulation with gonadotropins/ gonadotropin releasing factor agonists (a-GnRH), largely used currently for infertility treatment, can induce hepatic effects, demonstrated only in animals or in women with hyperstimulation syndrome. AIM: We wanted to estimate the number of included patients for which the variation of gamma-glutamyl-transferase (GGT) during and due to ovarian stimulation could be sustained by statistic validation. METHOD: In 23 consecutive patients, aged 23-45 years (mean 32.6 +/- 7.4 years) included in a fertility program, busereline, an a-GnRH was started the first day of the cycle and followed from the 14th ay with human menopausal gonadotropin. Ovulation was triggered with human chorionic gonadotropin. GGT was measured in the serum the first day (control), in the 14th, 19th, 24th day, the day before the triggering of the ovulation and two days after that. The statistic study used a distribution analysis (Student t test)--BMDP, SAS 6.0 and EpiInfo 5 software and calculated the necessary number of measurements in order to obtain significant (95%) variation for GGT (actual and = 5%). RESULTS AND CONCLUSIONS: The proper number of determinations which statistically support a possible significant difference is 29-38 and 46 for a 5% difference. The considered suppositions are able to support a correct estimation. PMID- 17410290 TI - Update on the management of refractory coeliac disease. AB - True Refractory Coeliac Disease (RCD) is being currently defined as persisting or recurring villous atrophy with crypt hyperplasia and increased intraepithelial lymphocytes in spite of a strict gluten free diet for more than 12 months or when severe persisting symptoms necessitate intervention independent of the duration of the dietary therapy. Currently two categories of RCD are being recognized: type I without aberrant T-cells and type II with aberrant T-cells detected by immunophenotyping by flowcytometric analysis or immunohistology of the intestinal mucosa. Establishing the diagnosis of RCD requires exclusion of other causes of villous atrophy and malignancies that may complicate coeliac disease. In contrast to patients with a high percentage of aberrant T cells, patients with RCD type I seem to profit from an immunosuppressive treatment. In cases of RCD II with persistent clinical symptoms and/or high percentage of aberrant T cells in intestinal biopsies in spite of a corticosteroid treatment, more aggressive therapeutic schemes should be considered. However, no therapy seems to be curative in RCD II. Cladribine (2-CDA) seems to have some role in the management of these patients. More recently, high dose chemotherapy followed by autologous stem cell transplantation has been used in patients resulting in a dramatic improvement in the clinical, laboratory, histopathological and immunological parameters. This review provides an overview of the currently available diagnostic and therapeutic methods in a complicated form of coeliac disease. PMID- 17410291 TI - Extrahepatic manifestations of chronic HCV infection. AB - Several extrahepatic manifestations have been reported in the natural history of hepatitis C virus infection (HCV). Up to 40-74% of patients infected with HCV might develop at least one extrahepatic manifestation during the course of their disease. Mixed Cryoglobulinemia (MC) is the most known and studied syndrome associated with HCV infection. It is a systemic vasculitis that may involve the skin, kidney and nervous system. A frequent reported association is that between HCV infection and non-Hodgkin lymphoma. The cryoglobulinemia may be the intermediary disorder, in fact some persistent forms of cyoglobulinemia can switch over to a more aggressive haematologic disorder. As compared to cutaneous vasculitis described in MC, HCV infection has been associated with dermatological disorders such as porphyria cutanea tarda and lichen planus. Thyroid disease (usually hypothyroidism) is commonly seen in people with HCV. Up to 25% have thyroid antibodies. Several studies described a correlation between HCV and lympho-cytic sialoadenitis, similar to sialoadenitis associated with idiopathic Sjogren syndrome, but we can define as "pseudo- Sjogren syndrome" the one associated with HCV infection, because it shows several differences in the idiopathic form. In the course of chronic HCV infection, a common obser-vation are rheumatological symptoms such as polyarthritis. The clinical pattern of joint involvement in the course of HCV infection varies from a rheumatoid arthritis like form (very rare), to a non erosive oligoarthritis involving the large-sized and middle joints. PMID- 17410292 TI - The applicability of radial endoscopic ultrasonography in pancreatic diseases. AB - Radial endoscopic ultrasonography (EUS) is used in pancreatic pathology for diagnosing such diseases as: chronic pancreatitis, cancer of the pancreas, neuroendocrine tumors, and the non-inflammatory cyst lesions of the pancreas. For diagnosing chronic pancreatitis, the use of more than four criteria of endoscopic diagnosis offers a high diagnostic accuracy. In the case of tumors of the pancreas, EUS detects small-size tumors, having a major diagnostic value in the preoperative staging and in assessing the tumor resectability. EUS can best identify insulinoma, while for the differential diagnosis of cystic neoplastic lesions data provided by radial EUS are not sufficient. In this respect, intracyst fluid analysis is required to identify malignant lesions or those having malignancy potential. PMID- 17410293 TI - Evaluating the liver tumors using three-dimensional ultrasonography. A pictorial essay. AB - The liver tumors represent a major public health issue. Among the imaging techniques, ultrasonography remains an important diagnostic method, taking into consideration its large availability, the non-invasive and repetitive characteristics and the low cost. Its current limits can be surpassed, at least at some extent, by using some new techniques such as the 3D ultrasonography. 3D ultra-sonography provides several advantages: increases the efficiency of the liver tumor screening by using the multislice function, allows a better time and human resource management, the precise location on three axes of the tumor and the exact assessment of the distances relative to the main vessel structures of the liver. It is also able to characterize the tumor texture and to sum up the total volume of liver masses, with applications in post-chemotherapy follow-up. PMID- 17410294 TI - Celiac artery compression syndrome: successful utilization of robotic-assisted laparoscopic approach. AB - Median arcuate ligament (MAL) syndrome, also known as the celiac axis compression syndrome (CACS) is rare, and a topic of ongoing academic controversy. CACS is a diagnosis of exclusion, characterized by the clinical triad of postprandial abdominal pain, weight loss, and vomiting. The classic management of CACS involves the surgical division of the MAL fibers. We report successful treatment of a 23-year-old woman with CACS utilizing the da Vinci Surgical System (Intuitive Surgical, Sunnyvale, California) via robotic-assisted minimally invasive surgical division of the MAL. To our knowledge this is the first report of this modality used in the treatment of the celiac axis compression syndrome. PMID- 17410295 TI - A case of portal cavernoma--associated essential thrombocythemia. AB - Portal cavernoma is a rare complication of portal vein thrombosis and may be a component of the myeloproliferative hematologic disorders. In the present case report, although the symptoms induced by portal hypertension and portal biliopathy had a concordant evolution with the portal cavernoma, the peripheral thrombocytosis and the medullar expression of essential thrombocythemia became relevant at 18 months. PMID- 17410296 TI - Problems in diagnosing lymphoma of the pancreas with computed tomography. A case report. AB - Primary lymphoma of the pancreas is a rare form of extranodal lymphoma accounting for less than 0.5% of pancreatic tumors. Percutaneous fine-needle aspiration of the pancreas with histopathological examination and immunohistochemical assay confirm the diagnosis. A 73 year old male presented with recurrent pancreatic type abdominal pain with significant weight loss over 1 year. He was pale with ill defined epigastric mass. Contrast enhanced CT showed an ill defined poorly marginated non enhancing hypodense mass lesion involving the body of the pancreas. CA 19-9 was normal. CT guided aspiration cytology was suggestive of hemolymphoid malignancy. Immunohistochemistry was positive for Leukocyte Common Antigen and CD 34. It was negative for CD3 and CD 20 indicating an undifferentiated lymphoma. Patient received two sessions of chemotherapy and was followed-up. PMID- 17410297 TI - Natural evolution of an intraductal papillary mucinous neoplasm of the pancreas. A case report. AB - Intraductal papillary mucinous neoplasms include a large spectrum of lesions communicating with the Wirsung duct, having a variable invasiveness from benign or borderline, to malignant (carcinoma in situ and invasive cancer). Final diagnosis is based on endoscopic ultrasound (EUS)-guided fine needle aspiration and histopathologic exam of surgical specimens. We present the case of a 28-year old woman, with several episodes of acute recurrent pancreatitis in the past 6 months, admitted for dyspepsia, nausea and loss of appetite. Imaging studies (transabdominal ultrasonography, CT scanning, MR cholangiopancreatography) showed a macrocystic, multilocular, corporeal tumor, communicating with the retrograde dilated Wirsung duct. EUS revealed hypoechoic material inside the cysts, raising the suspicion of an intraductal papillary mucinous neoplasm. Diagnosis was confirmed by EUS-guided fine needle aspiration, which found columnar mucinous cells within a mucin-rich fluid. The imaging evaluation was repeated after two years, showing a rapid evolution of the tumor. The patient refused surgical exploration and caudal pancreatectomy. In the context of the absence of clinical symptoms, the indolent evolution of these tumors and the excellent prognosis after resection, we consider that early identification and regular follow-up by EUS with fine needle aspiration is imperative, especially because of the limited success of other imaging methods. PMID- 17410298 TI - Unusual presentation and complication of caustic ingestion. Case report. AB - Caustic substances cause tissue destruction through liquefaction or coagulation reactions and the intensity of destruction depends on the type, concentration, time of contact and amount of the substance ingested. We report an unusual presentation and complication of caustic ingestion in a patient, who accidentally ingested sodium hydroxide. Our patient presented respiratory failure soon after admission and developed necrotizing esophagitis with progression to esophageal stenosis, which required surgical treatment. The complications were related to the amount of caustic soda ingested. PMID- 17410299 TI - A pyogenous gastric abscess that developed following ingestion of a piece of a wooden skewer: successful treatment with endoscopic incision. AB - A 62-year-old man with a medical history of duodenal ulcer was referred to our department for endoscopy, because of epigastralgia associated with mild anorexia. At endoscopy, a large protruding lesion with the appearance of a submucosal tumor was present at the posterior wall of the gastric body. Trying to perform an aspiration needle biopsy for cytological study, we observed the outflow of pus from the puncture point. An incision, 1 cm in length, was performed via a needle knife sphincterotomy, which led to drainage of the abscess and disappearance of symptoms. A 2.5 cm long piece of a wooden skewer was found to be embedded into the area of the bulge and was retrieved. PMID- 17410300 TI - Portal vein flow restoration and hepatic encephalopathy correction by ligation of a massive splenorenal shunt and splenectomy during Hassab-Paquet procedure. PMID- 17410301 TI - Pattern of relapses in Romanian patients with ulcerative colitis. PMID- 17410304 TI - The hygroscopic behaviour of individual aerosol particles in nickel refineries as investigated by environmental scanning electron microscopy. AB - The hygroscopic properties of individual aerosol particles (1-35 microm equivalent projected area diameter) from the Roasting, Anode Casting and Electrorefining Departments of two Ni refineries were studied by environmental scanning electron microscopy (ESEM) and energy-dispersive X-ray microanalysis (EDX) at a relative humidity of 96-98% (at a temperature of 5 degrees C). In the Roasting and Anode Casting Departments, most particles (60-85% by number) showed no visible change in size or surface morphology when exposed to high relative humidity. Approximately 15-30% of the particles developed a thin water film (growth factors between 1.006 and 1.06) indicating the presence of thin surface coatings of sulfates. About 10% of the particles in the Roasting Department formed droplets (growth factors between 1.1 and 2.6) which always contained a large portion of insoluble material. In the Electrorefining Department, most particles (approximately 60%) were residues from the electrolysis bath solution. At a relative humidity of 96-98% these particles formed a solution which contained only small insoluble inclusions. About 30% of the particles in the Electrorefining Department developed thin water films. As only a small fraction of the particles increased substantially in size when exposed to high relative humidity, the deposition pattern of the total aerosol mass fraction will not be changed substantially by hygroscopic growth. The frequent occurrence of thin surface coatings of soluble material on insoluble Ni compounds has to be considered for health assessment purposes. PMID- 17410305 TI - Determination of six anti-infectives in wastewater using tandem solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - A rugged and specific method based on tandem solid-phase extraction and liquid chromatography-tandem mass spectrometry for the determination of anti-infectives in raw sewage and wastewater plant effluents was developed. Analyte recoveries from spiked effluents ranged from 68 to 104%. Two specific selected reaction monitoring transitions and their peak area ratios were used to avoid false positives and confirm the presence of the targeted substances. Detection limits allowed low nanogram per litre detection (0.3-22 ng L(-1)). The method was successfully applied to real samples from the Montreal wastewater treatment plant. All the studied anti-infectives were found in the wastewater samples in concentrations ranging from 39 to 276 ng L(-1). Mean flows of anti-infectives were estimated from effluent concentrations and it was found that large amounts (>118 g day(-1) up to 830 g day(-1)) are discharged in the receiving waters of the St Lawrence River. PMID- 17410306 TI - Dehalogenation of polychlorinated biphenyls and polybrominated diphenyl ethers using a hybrid bioinorganic catalyst. AB - The environmentally prevalent polybrominated diphenyl ether (PBDE) #47 and polychlorinated biphenyls (PCBs) #28 and #118 were challenged for 24 hours with a novel biomass-supported Pd catalyst (Bio-Pd(0)). Analysis of the products via GC MS revealed the Bio-Pd(0) to cause the challenged compounds to undergo stepwise dehalogenation with preferential loss of the least sterically hindered halogen atom. A mass balance for PCB #28 showed that it is degraded to three dichlorobiphenyls (33.9%), two monochlorobiphenyls (12%), and biphenyl (30.7%). The remaining mass was starting material. In contrast, while PCB #118 underwent degradation to yield five tetra- and five trichlorinated biphenyls, no less chlorinated products or biphenyl were detected, and the total mass of degraded products was 0.3%. Although the Bio-Pd(0) material was developed for treatment of PCBs, a mass balance for PBDE #47 showed that the biocatalyst could prove a potentially useful method for treatment of PBDEs. Specifically, 10% of PBDE #47 was converted to identifiable lower brominated congeners, predominantly the tribrominated PBDE #17 and the dibrominated PBDE #4, 75% remained intact, while 15% of the starting mass was unaccounted for. PMID- 17410307 TI - Classification of alluvial soils according to their potential environmental risk: a case study for Belgian catchments. AB - Alluvial soils may represent important sinks of contaminants as a result of the deposition of contaminated sediments along the river by overbank flooding or after dredging. Because of the erosion of alluvial deposits or the release of contaminants from sediments, alluvial soils can also be a source of contamination. In this paper, a risk assessment for contaminated (alluvial) soils is presented. The approach, mainly based on physico-chemical soil characteristics, single extractions and leaching tests, is illustrated by means of a case study from four Belgian catchments. The extractions and leaching tests that were used have been validated by European testing programs and can provide valuable information for classifying the potential environmental risks of soils. Irrespective of the location, pH, organic carbon content and 'mobilisable' metal concentrations were the most important factors explaining 'mobile' metal concentrations in the alluvial soils. Additionally, the data of the physico chemical soil characterization, extractions and leaching tests were combined with local and regional factors to classify the alluvial soils in different categories according to their actual and potential risk for the environment. PMID- 17410308 TI - The role of humic non-exchangeable binding in the promotion of metal ion transport in groundwaters in the environment. AB - Metal ions form strong complexes with humic substances. When the metal ion is first complexed by humic material, it is bound in an 'exchangeable' mode. The metal ion in this fraction is strongly bound, however, if the metal-humic complex encounters a stronger binding site on a surface, then the metal ion may dissociate from the humic substance and be immobilised. However, over time, exchangeably-bound metal may transfer to a 'non-exchangeable' mode. Transfer into this mode and dissociation from it are slow, regardless of the strength of the competing sink, and so immobilisation may be hindered. A series of coupled chemical transport calculations has been performed to investigate the likely effects of non-exchangeable binding upon the transport of metal ions in the environment. The calculations show that metal in the non-exchangeable mode will have a significantly higher mobility than that in the exchangeable mode. The critical factor is the ratio of the non-exchangeable first-order dissociation rate constant and the residence time in the groundwater column, metal ion mobility increasing with decreasing rate constant. A second series of calculations has investigated the effect of the sorption to surfaces of humic/metal complexes on the transport of the non-exchangeably bound metal. It was found that such sorption may reduce mobility, depending upon the humic fraction to which the metal ion is bound. For the more weakly sorbing humic fractions, under ambient conditions (humic concentration etc.) the non exchangeable fraction may still transport significantly. However, for the more strongly sorbed fractions, the non-exchangeable fraction has little effect upon mobility. In addition to direct retardation, sorption also increases the residence time of the non-exchangeable fraction, giving more time for dissociation and immobilisation. The non-exchangeable dissociation reaction, and the sorption reaction have been classified in terms of two Damkohler numbers, which can be used to determine the importance of chemical kinetics during transport calculations. These numbers have been used to develop a set of rules that determine when full chemical kinetic calculations are required for a reliable prediction, and when equilibrium may be assumed, or when the reactions are sufficiently slow that they may be ignored completely. PMID- 17410309 TI - Indoor and outdoor concentrations of PM2.5 trace elements at homes, preschools and schools in Stockholm, Sweden. AB - Fine particles (PM2.5) were sampled indoors and outdoors at 40 sampling sites; in ten classrooms in five schools, at ten preschools and 20 non-smoking homes, in three communities in Stockholm, Sweden, during nine 2-week periods. Each sampling site was sampled twice, once during winter and once during spring. The samples were analysed for elemental concentrations using X-ray fluorescence (XRF) spectroscopy. In all locations significantly higher outdoor concentrations were found for elements that are related to long-range transported air masses (S, Ni, Br and Pb), while only Ti was higher indoors in all locations. Similar differences for S, Br and Pb were found in both seasons for homes and schools. In preschools different seasonal patterns were seen for the long-range transported elements S, Br and Pb and the crustal elements Ti, Mn and Fe. The indoor/outdoor ratios for S and Pb suggest an outdoor PM2.5 particle net infiltration of about 0.6 in these buildings. The community located 25 km from the city centre had significantly lower outdoor concentrations of elements of crustal or traffic origin compared with the two central communities, but had similar levels of long range transported elements. Significant correlations were found between PM2.5 and most elements outdoors (rs = 0.45-0.90). Copper levels were found to correlate well (rs = 0.64-0.91) to the traffic marker NO2 during both winter and spring in all locations. Copper may be a suitable elemental marker for traffic-related aerosols in health studies in areas without other significant outdoor Cu sources. PMID- 17410310 TI - Speciation of organotins in polyvinyl chloride pipe via X-ray absorption spectroscopy and in leachates using GC-PFPD after derivatisation. AB - Three different polyvinyl chloride (PVC) pipe types were subjected to de-ionized water exposures over the course of at least 180 days. Water exposed to the pipe was analyzed for organotin speciation and concentration. Organotin concentrations were the highest during the first 1-5 days. The species and concentrations of organotins leached varied by pipe type. Data were normalized by surface area in order to compare laboratory results with results from a residential pipe system. For one pipe type, the lowest non-zero concentrations from the laboratory tests overestimated organotin concentrations in solution when compared with water samples from the same pipe type in a residence. For organotin exposure estimates, a range of 0.1 ng m(-2) to 10 ng m(-2) could be used for mature pipes (e.g. in use for 1 year). These estimates should be refined with more field study, however, due to the high variation in organotin species and concentrations leached as a function of pipe type, accuracy within an order of magnitude may be optimal as, in many instances, the type of pipe installed or buried may be unknown. X-ray absorption spectroscopy (XAS) was used to identify organic and inorganic tin species in reference materials and the PVC samples. Monobutyl tin was identified as the primary organotin species in the pipes. Results from the XAS analyses also indicate that the technique shows promise for distinguishing between inorganic tin and organotins. Furthermore, organotins may be distinguished between mono-, di-, and tri-ligand species using XAS. PMID- 17410311 TI - Highway runoff quality in Ireland. AB - Highway runoff has been identified as a significant source of contaminants that impact on the receiving aquatic environment. Several studies have been completed documenting the characteristics of highway runoff and its implication to the receiving water in the UK and elsewhere. However, very little information is available for Ireland. The objective of this study was to determine the quality of highway runoff from major Irish roads under the current road drainage design and maintenance practice. Four sites were selected from the M4 and the M7 motorways outside Dublin. Automatic samplers and continuous monitoring devices were deployed to sample and monitor the runoff quality and quantity. More than 42 storm events were sampled and analysed for the heavy metals Cd, Cu, Pb, and Zn, 16 US EPA specified PAHs, volatile organic compounds including MTBE, and a number of conventional pollutants. All samples were analysed based on the Standard Methods. Significant quantities of solids and heavy metals were detected at all sites. PAHs were not detected very often, but when detected the values were different from quantities observed in UK highways. The heavy metal concentrations were strongly related to the total suspended solids concentrations, which has a useful implication for runoff management strategies. No strong relationship was discovered between pollutant concentrations and event characteristics such as rainfall intensity, antecedent dry days (ADD), or rainfall depth (volume). This study has demonstrated that runoff from Irish motorways was not any cleaner than in the UK although the traffic volume at the monitored sites was relatively smaller. This calls for a site specific investigation of highway runoff quality before adopting a given management strategy. PMID- 17410312 TI - Volatile organic compound exposure of suburban elementary students in Taiwan. PMID- 17410313 TI - Kinetic and mechanistic study of photodechlorination of 2,2',4,4' tetrachlorobiphenyl in surfactant solutions. PMID- 17410314 TI - Zinc and copper toxicity to soil bacteria and fungi from zinc polluted and unpolluted soils: a comparative study with different types of Biolog plates. PMID- 17410315 TI - Isolation and characterization of a nitrobenzene degrading Streptomyces strain from activated sludge. PMID- 17410316 TI - Initial survey of plasma vitellogenin and gonadal development in male carp (Cyprinus carpio) from three locations in New Jersey, USA. PMID- 17410318 TI - Response to P. Hilton: of porcupines and poodles-a joint challenge to industry and the profession. PMID- 17410319 TI - POEMS syndrome complicated by extensive arterial thromboses. AB - POEMS syndrome--characterised by polyneuropathy, organomegaly, endocrinopathy, monoclonal protein and skin changes--is a rare multi-systemic disease. Its skin change is often like that of connective tissue diseases such as scleroderma. Although clinical manifestations of POEMS syndrome are known to be diverse, arterial or venous thrombosis is a less-recognised feature. We report a 41-year old man with scleroderma-like skin change who was initially negative for monoclonal protein, but finally confirmed as having POEMS syndrome. During the disease course, he was complicated by extensive arterial thromboses including stroke. This case evokes the need to include POEMS syndrome in the differential diagnoses in patients with scleroderma-like skin change and to repeat measurements of monoclonal protein to confirm its diagnosis. A POEMS patient with extensive arterial thromboses may have a catastrophic disease course, requiring earlier diagnosis and more aggressive treatment. PMID- 17410320 TI - Anti-CCP antibodies have more diagnostic impact than rheumatoid factor (RF) in a population tested for RF. AB - To compare the diagnostic powers of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) in a population selected for its high statistical relevance, over a 6-month period, an informed consent to test for anti-CCP was obtained from 1,025 consecutive patients for whom RF was ordered at a University laboratory. Within 1 year, a diagnosis was obtained without informing the physician about the anti-CCP result. Extensive statistical analyses were performed. A total of 768 patients satisfied the inclusion criteria, and 132 were classified as having RA, yielding a pre-test probability of RA of 17%. The sensitivities for anti-CCP and RF were 62 and 64% (P = 0.83), and the specificities were 97 and 90% (P < 0.001), respectively. The positive predictive value (PPV) was 79% for anti-CCP and 56% for RF (P < 0.001), whereas the negative predictive value was 92% for both. The likelihood ratio (LR) was 17.9 for anti CCP and 6.2 for RF (P < 0.005). Forty RA patients were diagnosed with RA of less than 2 years length, and the same significant statistic differences between anti CCP and RF were observed. Placing the results of both tests together, or using different cutoff points, increased the diagnostic utility of the tests. The anti CCP test has statistically shown significant higher specificity, PPV, and LR for RA than the RF test in a clinically diverse population. If new criteria are to be devised to help diagnose early RA, anti-CCP should be included because it has a greater diagnostic impact than RF. PMID- 17410321 TI - Interaction between estrogen receptor-alpha and butyrylcholinesterase genes modulates Alzheimer's disease risk. PMID- 17410322 TI - Sporadic myopathy and exercise intolerance associated with the mitochondrial 8328G>A tRNALys mutation. PMID- 17410323 TI - Small-fiber neuropathy and the 3243A>G mutation in mitochondrial DNA. PMID- 17410324 TI - A novel mutation in GCH-1 gene in a case of dopa-responsive dystonia. PMID- 17410325 TI - Beneficial effects of 3,4-diaminopyridine on positioning downbeat nystagmus in a circumscribed uvulo-nodular lesion. PMID- 17410326 TI - Paroxysmal cervicobrachial cough-induced pain in a patient with syringomyelia extending into spinal cord posterior gray horns. PMID- 17410327 TI - Nucleosomes as a new prognostic marker in early cerebral stroke. AB - BACKGROUND: The prognostic relevance of blood markers in cerebral stroke is still a matter of controversial debate. PATIENTS AND METHODS: In sera of 63 patients, nucleosomes, neuronspecific enolase (NSE), S100 protein, and C-reactive protein (CRP) were determined daily during the first week after cerebral stroke. Infarction volume was quantified by CT or MRI and the clinical status by Barthel Index (BI) at admission, discharge, and after 12 months (prognosis). All markers were evaluated by univariate and multivariate analysis on their prognostic relevance. RESULTS: During observation time (12 months), three patients died and 33 reached complete recovery. Infarction volume, nucleosomes, NSE, S100, and CRP correlated significantly with clinical status at admission. The same markers except CRP and initial BI correlated with recovery after 12 months. Almost all patients with initial BI double dagger 50 reached complete recovery. In patients with initially severe defects (BI < 50), nucleosomes and S100, both at day 3, were found to be prognostically relevant. At 100%-specificity for non-recovery, only nucleosomes maintained their prognostic power (sensitivity 52.6%; p = 0.014), whereas S100 did not (sensitivity 16.7%; p = 0.25). In multivariate analysis, nucleosomes and BI at admission showed independent prognostic relevance (p = 0.039). CONCLUSION: Circulating nucleosomes and clinical scores provide independent prognostic information concerning the later outcome in patients with initially severe defects after stroke. PMID- 17410328 TI - Deep brain stimulation of the nucleus accumbens and the internal capsule in therapeutically refractory Tourette-syndrome. PMID- 17410329 TI - Dysarthria and paresthesia of the thumb caused by a small cortical infarction. PMID- 17410330 TI - Metabolic deactivation of low-grade glioma during chemotherapy. PMID- 17410331 TI - Structure-activity investigations of polyamine-anthracene conjugates and their uptake via the polyamine transporter. AB - A series of polyamine conjugates were synthesized and evaluated for their ability to target the polyamine transporter (PAT) in two Chinese hamster ovary (CHO) cell lines (PAT-active CHO and PAT-inactive CHOMG). This systematic study identified salient features of the polyamine architecture required to target and enter cells via the PAT. Indeed, the separation of charges, the degree of N-alkylation, and the spacer unit connecting the N(1)-terminus to the appended cytotoxic component (anthracene) were found to be key contributors to optimal delivery via the PAT. Using the CHO screen, the homospermidine motif (e.g., 4,4-triamine) was identified as a polyamine vector, which could enable the selective import of large N(1)-substituents (i.e., naphthylmethyl, anthracenylmethyl and pyrenylmethyl), which were cytotoxic to cells. The cell selectivity of this approach was demonstrated in B-16 murine melanoma cells and normal melanocytes (Mel-A). Three polyamine areas (recognition and transport, vesicle sequestration and polyamine-target interactions) were identified for future research. PMID- 17410332 TI - Antizyme and antizyme inhibitor activities influence cellular responses to polyamine analogs. AB - Close structural analogs of spermidine and spermine, polyamine mimetics, are potential chemotheraputic agents as they depress cellular polyamines required for tumor growth. Specific mimetic analogs stimulate synthesis of the regulatory protein antizyme (AZ), which not only inactivates the initial enzyme in polyamine biosynthesis but also inhibits cellular uptake of polyamines. The role of AZ induction in influencing cellular uptake of representative analogs was investigated using three analogs produced by Cellgate Inc., CGC-11047, CGC-11102, and CGC-11144, which exhibit markedly distinct AZ-inducing potential. An inverse correlation was noted between the AZ-inducing activity of a compound and the steady-state levels accumulated in cells. As some tumor cells over express AZI as a means of enhancing the polyamines required for aggressive growth, analog sensitivity was examined in transgenic CHO cells expressing exogenous antizyme inhibitor protein (AZI). Although AZI over expression increased cell sensitivity to analogs, the degree of this affect varied with the analog used. PMID- 17410333 TI - Oxidative stress and inflammation in the pathogenesis of activated polyamine catabolism-induced acute pancreatitis. AB - The markers of oxidative stress and inflammation were studied in acute pancreatitis in transgenic rats exhibiting activated polyamine catabolism. In addition, the effect of bismethylspermine (Me(2)Spm) pretreatment, preventing pancreatitis in this model, on these mediators was investigated. Lipid peroxidation was increased at 6 and 24 h after induction of pancreatitis. These changes as well as the markedly decreased superoxide dismutase activity at 24 h were abolished by Me(2)Spm pretreatment. Glutathione level and catalase activity changed transiently, and the effect of Me(2)Spm was clear at 24 h. Serum inflammatory cytokine levels increased already at 4 h whereas NF-kappaB was distinctly activated only at 24 h. Me(2)Spm prevented the increase in TNF-alpha and IL-6 while it had no effect on NF-kappaB activation. These results show that typical inflammatory and, to a lesser degree, some oxidative stress mediators are involved and beneficially affected by the disease-ameliorating polyamine analogue in our pancreatitis model. PMID- 17410334 TI - Association of telomerase gene hTERT polymorphism and malignant gliomas. AB - BACKGROUND: The MNS16A polymorphism is located in the downstream region of the hTERT gene and affects telomerase activity. METHODS: MNS16A has been investigated as a potential risk factor and/or prognostic marker for malignant glioma in a cohort of 352 patients (205 glioblastoma, 147 anaplastic gliomas) and 305 controls. RESULTS: The S ("short") allele (which results in a higher telomerase activity) was significantly more frequent in glioma patients compared to the control population (278/704=39.5% vs. 200/610=32.8%; P=0.012). The odd ratios were 1 for LL (taken as reference), 1.33 [0.96; 1.84] for SL and 2.05 [1.22; 3.44] for SS. However, in contrast to a previous report, no significant difference of survival was found between SS, LL and SL allelotypes. CONCLUSION: We found here the short allele of MNS16A more frequent in glioma patients, but it did not seem to be predictive of survival. PMID- 17410335 TI - Expression of human MCM6 and DNA Topo II alpha in craniopharyngiomas and its correlation with recurrence of the tumor. AB - Craniopharyngioma often recurs after resection resulting in poor outcome for the affected patients. The reliable criteria for predicting the tumor behavior are still lacking. It has been suggested that proliferative potential of the tumor cells is necessary for recurrence. Present study evaluated the cell multiplication activity, which is possibly related to relapse in 32 patients with adamantine epithelioma (AE) and 31 patients with squamous papillary tumor (SP). For this tissue specimens from their primary and recurring tumors were collected and immunohistochemical analysis of the expression of minichromosome maintenance protein 6 (MCM6) and DNA topoisomerase II alphalpha (DNA Topo II alpha) in the tumor sections was performed by quantitative microscopy. It was found that the MCM6 label index (LI) was significantly higher in AE than that of SP, and in the primary tumors of both subtypes the LIs of tumors with recurrence were higher than those without recurrence. There was a strong linear positive correlation between MCM6 LI and DNA Topo II alpha LI (r = 0.713; p = 0.000) in the craniopharyngiomas studied. The median MCM6 LI of the total 20 recurrent craniopharyngiomas (31.49%) was not significantly different from that of their primary tumors (29.65%). The long term risk of tumor recurrence is higher in AE than SP and it is associated with MCM6 and DNA Topo II alpha expression. PMID- 17410336 TI - Unusual case of aortic valve involvement in patient with Loffler's endomyocarditis: management, follow-up and short review of the literature. AB - Idiopathic hypereosinophilic syndrome (IHES) is an uncommon systemic disease which is characterised by blood eosinophilia and multiple clinical presentations. Cardiac involvement is the major cause of mortality and morbidity. Here we describe a 59-year-old man with symptoms of progressive dyspnea on exertion, and productive cough as an unusual case of Loffler endomyocarditis with a mass on the aortic valve which showed regression with treatment. PMID- 17410337 TI - Imatinib-induced optic neuritis in a patient of chronic myeloid leukemia. AB - Imatinib is presently the commonest prescribes drug for patients of chronic myelogenous leukemia (CML) in chronic phase (CP) worldwide. Its ocular side effects are little known. Optic neuritis caused by this drug has not been reported (Medline search). We describe perhaps the first case of optic neuritis caused by this drug. PMID- 17410338 TI - Acetaldehyde mediates growth stimulation of ethanol-stressed Saccharomyces cerevisiae: evidence of a redox-driven mechanism. AB - The ability of acetaldehyde (90 mg l(-1)) to stimulate ethanol-stressed S. cerevisiae fermentations is examined and reasons for the effect explored. Alternative metabolic electron acceptors generated similar stimulatory effects to acetaldehyde, decreasing the ethanol-induced growth lag phase from 9 h to 3 h, suggesting a redox-driven effect. The exposure to ethanol caused an instant 60% decline in intracellular NAD(+) which was largely prevented by the addition of acetaldehyde. Furthermore, the exposure to ethanol affected glycolysis by decreasing the rate of glucose utilisation from 0.33 g glucose g(-1) biomass h( 1) to 0.11 g glucose g(-1) biomass h(-1), while the addition of acetaldehyde to an ethanol stressed culture increased this rate to 0.14 g glucose g(-1) biomass h(-1). PMID- 17410339 TI - Anaerobic biotechnological approaches for production of liquid energy carriers from biomass. AB - In recent years, increasing attention has been paid to the use of renewable biomass for energy production. Anaerobic biotechnological approaches for production of liquid energy carriers (ethanol and a mixture of acetone, butanol and ethanol) from biomass can be employed to decrease environmental pollution and reduce dependency on fossil fuels. There are two major biological processes that can convert biomass to liquid energy carriers via anaerobic biological breakdown of organic matter: ethanol fermentation and mixed acetone, butanol, ethanol (ABE) fermentation. The specific product formation is determined by substrates and microbial communities available as well as the operating conditions applied. In this review, we evaluate the recent biotechnological approaches employed in ethanol and ABE fermentation. Practical applicability of different technologies is discussed taking into account the microbiology and biochemistry of the processes. PMID- 17410340 TI - [Development of the concept of identity in psychiatry and philosophy]. AB - The present paper discusses possible solutions to the problem of personal identity and intends to demonstrate criteria enabling us to view a person as the same at different time points in his life. Of special interest are the so-called memory criterion and the criterion of physical identity/continuity: a person remains the same if he is able to remember his past "states" or if he continues to have his former body (brain). The philosophical discussion of the last decades made clear that all attempts to reduce personal identity to memory or physical continuity are contradictory. In this paper we propose regarding personal identity as a normative phenomenon that evades a purely descriptive or neurobiological approach. PMID- 17410341 TI - [Working memory dysfunctions in psychiatric disorders]. AB - Working memory disturbances occur in various psychiatric disorders and play a major role for general cognitive ability in everyday life and thus social integration of the patients. In schizophrenia and bipolar disorder, working memory deficits have been found to be associated with certain genotypes and clinical symptoms. This makes them promising endophenotypes, i.e., mediators between genotype and phenotype. Furthermore recent studies indicate that the identification of selective deficits in working memory may be useful in order to define patient subgroups that are more homogenous with respect to the functional integrity or dysfunction of neural systems underlying working memory subcomponents. This scientific approach may ultimately lead to better understanding of the pathophysiologic mechanisms underlying working memory dysfunctions in psychiatric disorders and may provide clinicians with additional information for diagnostic and therapeutic decisions. PMID- 17410342 TI - Lipopolysaccharide-binding protein, lipopolysaccharide, and soluble CD14 in sepsis of critically ill neonates and children. AB - OBJECTIVE: To compare the diagnostic accuracy of lipopolysaccharide-binding protein (LBP) for sepsis in critically ill neonates and children with the two markers participating in the same inflammatory pathway, lipopolysaccharide and soluble CD14. DESIGN AND SETTING: Prospective, observational study in a multidisciplinary neonatal and pediatric intensive care unit. PATIENTS: 47 critically ill neonates and 49 critically ill children with systemic inflammatory response syndrome (SIRS) and suspected sepsis, classified into two groups: those with and those without sepsis. INTERVENTIONS: Serum LBP, lipopolysaccharide, soluble CD14, C-reactive protein, and procalcitonin were measured on 2 consecutive days. The area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and predictive values were evaluated. RESULTS: AUC for LBP on the first day of suspected infection was 0.97 in neonates aged under 48 h, 0.93 in neonates over 48 h and 0.82 in children. AUCs for lipopolysaccharide and soluble CD14 were 0.77 and 0.74 in neonates under 48 h, 0.53 and 0.76 in neonates over 48 h, and 0.72 and 0.53 in children. AUCs for procalcitonin and C-reactive protein were 0.65 and 0.89 in neonates under 48 h, 0.65 and 0.91 in neonates over 48 h, and 0.76 and 0.69 in children. CONCLUSIONS: In critically ill neonates and children LBP concentration on the first day of suspected sepsis is a better marker of sepsis than lipopolysaccharide, soluble CD14, procalcitonin, and in neonates younger than 48 h and children, also a better marker than C-reactive protein. Lipopolysaccharide and soluble CD14 are not suitable markers for the differentiation of infectious and noninfectious SIRS. PMID- 17410343 TI - End-of-life care in Brazil. PMID- 17410344 TI - Sepsis-associated delirium. AB - OBJECTIVE: Sepsis-associated delirium is a common and poorly understood neurological complication of sepsis. This review provides an update of the diagnostic criteria and treatment strategies and the current knowledge about the mechanisms involved in sepsis associated brain dysfunction. DATA SOURCES: Articles published between 1981 and 2006 were identified through a Medline search for "encephalopathy" and "sepsis" and by hand searching of articles cited in the identified publications. The immune response to sepsis results in multiorgan failure including brain dysfunction. DISCUSSION: The potential mechanisms for sepsis-associated delirium include vascular damage, endothelial activation, breakdown of the blood-brain barrier, metabolic disorders, brain inflammation and apoptosis. On the other hand, there is evidence for distinct neuroprotective factors, such as anti-inflammatory mediators and glial cell activity. CONCLUSIONS: The diagnosis of sepsis-associated delirium relies mainly on clinical and electrophysiological criteria, and its treatment is entirely based on general management of sepsis. PMID- 17410345 TI - Bone scintigraphy in patients with bipartite patella. AB - This study was performed to assess the diagnostic value of bone scintigraphy to differentiate symptomatic bipartite patella from asymptomatic bipartite patella. Thirty-seven bipartite patellae (15 symptomatic and 22 asymptomatic) were evaluated by bone scintigraphy. Bone scintigraphic activity in bipartite patella was classified into the following four groups. Group A: Eleven bipartite patellae (10 symptomatic and 1 asymptomatic) demonstrated physiological and marked uptake at the epiphysio-metaphysial junction of the distal femur in bone scintiscans. Therefore, high scintigraphic uptake in the bipartite patella was difficult to distinguish from such uptake due to overlap. Group B: Five bipartite patellae (4 symptomatic and 1 asymptomatic) demonstrated both physiologically high scintigraphic uptake at the epiphysio-metaphysial junction of the distal femur and high scintigraphic uptake in the bipartite patella. Group C: Seventeen bipartite patellae (1 symptomatic and 16 asymptomatic) demonstrated abnormally high scintigraphic uptake in the bipartite patella. Group D: Four asymptomatic bipartite patellae did not demonstrate an abnormally high scintigraphic uptake. After excluding Group A, the proportion of positive bone scans in bipartite patella was 84.6% (22 of 26 patellae). Similarly, after excluding Group A, the proportion of positive bone scans in symptomatic bipartite patella was 100% (5 of 5 patellae) and in asymptomatic bipartite patella 81.0% (17 of 21 patellae). Statistical analysis using Fisher's exact test showed no significant differences in the proportion of positive bone scans between both groups (P = 0.5457). In conclusion, abnormally high scintigraphic uptake is frequent findings in both symptomatic and asymptomatic bipartite patella, and bone scintigraphy is not useful to differentiate between them. Therefore, surgical treatment should not be considered based only on scintigraphic findings. PMID- 17410346 TI - Disturbances in the arthroscopic view defined with video analysis. AB - Optimization of arthroscopic view with technological innovation. To formulate objective measures for arthroscopic image quality. This is achieved by defining uniquely interpretable descriptions of disturbances of the arthroscopic image. Disturbances in the arthroscopic view were defined focusing on the size of the disturbance area, and the duration of a disturbance. They were categorized as Bleeding, Turbidity, Air Bubbles, Loose Fibrous Tissue, Attached Fibrous Tissue, Tissue too Close, and Instrument too Close. Ten arthroscopic knee procedures were recorded on digital film with a frame rate of 25 images per second (almost 5 h in total). Using time-action analysis, two investigators individually scored the presence and type of each disturbance in all 5 h of the ten arthroscopies. The kappa statistic was used to assess agreement between the investigators. Additionally, five other observers, who had no prior training, scored 1 min of each of the ten arthroscopies (total of 10 min) to evaluate the definitions for unique interpretability. The adjusted kappa statistic (kappa is corrected for high prevalence) shows good tester agreement for all disturbances (range 0.71 0.96), also for the observers who had no prior training. Only Turbidity shows a moderate agreement (0.59 and 0.44). This is due to the difficulty to assess the exact start and end time of Turbidity. We succeeded in describing uniquely interpretable definitions for seven disturbances. This allows objective and quantitative determination of arthroscopic image quality, which is required to improve future arthroscopic equipment and training facilities. PMID- 17410347 TI - Mercury analysis. PMID- 17410348 TI - 'Gate effect' in templated polyacrylamide membranes influences the electrotransport of proteins and finds applications in proteome analysis. AB - Templating is an effective way for the structural modifications of a material and hence for altering its functional properties. Here protein imprinting was exploited to alter polymeric polyacrylamide (PAA) membranes. The sieving properties and selection abilities of the material formed were evaluated by studying the electrically driven transport of various proteins across templated PAA membranes. The sieving properties correlated with the templating process and depended on the quantity of template used during the polymerisation. For 1 mg/mL protein-templated membranes a 'gate effect' was shown, which induced a preferential migration of the template and of similar-size proteins. Such template preferential electrotransport was exploited for the selective removal of certain proteins in biological fluids prior to proteome analysis (depletion of albumin from human serum); the efficiency of the removal was demonstrated by analysing the serum proteome by two-dimensional electrophoresis experiments. PMID- 17410349 TI - Testing organic solvents for the extraction from fish of sulfophenylcarboxylic acids, prior to determination by liquid chromatography-mass spectrometry. AB - The present paper describes the use of different solvent mixtures to extract from fish various sulfophenylcarboxylic acids (SPCs of C(6) to C(13)), and their originating compounds, linear alkylbenzene sulfonates (LAS of C(10) to C(13)). The analytical method utilized involves pressurized liquid extraction, followed by preconcentration of the samples, purification by solid-phase extraction, and finally identification and quantification of the target compounds by high performance liquid chromatography-mass spectrometry using a system equipped with an electrospray interface operating in negative ion mode. The SPCs and LAS were extracted from spiked fish first with hexane to remove interference from fats, then with different mixtures of solvents: dichloromethane followed by methanol; 50:50 dichloromethane-methanol; and 30:70 dichloromethane-methanol. The LAS recoveries obtained with these three extraction options were high (between 68.5 and 80.8%); however, owing to the low percentages obtained for SPC homologues (13.5, 13.1, and 15.9%, respectively), another extraction procedure with methanol was developed in order to increase these recoveries. The percentage of recovery for total SPCs with the methanolic extraction was higher (90.1%), with a standard deviation of 9.9, and the LAS recoveries also increased (99.9%). Detection limits were between 1 and 22 ng g(-1) for LAS, and between 1 and 58 ng g(-1) for SPCs. Quantitation limits were between 4 and 73 ng g(-1) for LAS, and between 2 and 193 ng g(-1) for SPCs. This method has been applied to measure the biotransformation of 2OC(10) LAS (where O is a sulfophenyl group) in fish exposed in a flow-through system, and enabled the separation and identification of SPCs from 5OC(6) to 9OC(10). PMID- 17410350 TI - Investigation of polynuclear Zr(IV) hydroxide complexes by nanoelectrospray mass spectrometry combined with XAFS. AB - Polynuclear species of zirconium in acidic aqueous solution are investigated by combining X-ray absorption spectroscopy (XAFS) and nanoelectrospray mass spectrometry (ESI-MS). Species distributions are measured between pHC 0 and pHC 3 for [Zr]=1.5-10 mM. While the monomer remains a minor species, with increasing pH the degree of polymerization increases and the formation of tetramers, pentamers, octamers, and larger polymers is observed. The high resolution of the mass spectrometer permits the unambiguous determination of polynuclear zirconium hydroxide complexes by means of their isotopic patterns. The relative abundances of mononuclear and polynuclear species present simultaneously in solution are measured, even if one of the species contributes only 0.1% of the Zr concentration. For the first time it has been directly observed that the hydrolysis of polynuclear Zr species is a continuous process which leads to charge compensation through the sequential substitution of water molecules by hydroxide ligands until doubly charged polymers dominate at conditions (H+ and Zr concentrations) close to the solubility of Zr(OH)4(am). The invasiveness of the electrospray process was minimized by using very mild declustering conditions, leaving the polynuclear species within a solvent shell of approximately 20 water molecules. PMID- 17410351 TI - Spectrophotometric assay for detection of aromatic hydroxylation catalyzed by fungal haloperoxidase-peroxygenase. AB - Agrocybe aegerita peroxidase (AaP) is a versatile heme-thiolate protein that can act as a peroxygenase and catalyzes, among other reactions, the hydroxylation of aromatic rings. This paper reports a rapid and selective spectrophotometric method for directly detecting aromatic hydroxylation by AaP. The weakly activated aromatic compound naphthalene served as the substrate that was regioselectively converted into 1-naphthol in the presence of the co-substrate hydrogen peroxide. Formation of 1-naphthol was followed at 303 nm (epsilon (303) = 2,010 M(-1) cm( 1)), and the apparent Michaelis-Menten (K (m)) and catalytic (k (cat)) constants for the reaction were estimated to be 320 microM and 166 s(-1), respectively. This method will be useful in screening of fungi and other microorganisms for extracellular peroxygenase activities and in comparing and assessing different catalytic activities of haloperoxidase-peroxygenases. PMID- 17410352 TI - Biomedical imaging archive network. PMID- 17410353 TI - The prevalence of lumbar facet joint edema in patients with low back pain. AB - OBJECTIVE: The objective was to assess the prevalence of lumbar facet joint edema in patients with low back pain. MATERIALS AND METHODS: Lumbar spine MR examinations (1.5 T) of 145 consecutive patients (87 women, 58 men; mean age 52.8, range 17-94 years) were retrospectively evaluated with regard to the presence of facet joint edema. The MR protocol included sagittal short-tau inversion recovery (STIR), T1- and T2-weighted as well as transverse T2-weighted images. In 9 patients follow-up MR examinations were performed and results were compared with pain. The agreement between the change in intensity of facet joint edema and the change in intensity of pain was assessed using kappa statistics and Kendall's tau coefficient. RESULTS: In 21 of the 145 patients (14%) edema was found at the facet joints: in 52.4% at L4/5, in 19.0% at L5/S1, in 14.3% at L4/5 and L5/S1, in 9.5% at L3/4 and L4/5, and in 4.8% at L3/4. The agreement between the change in pain score and intensity of edema within the follow-up group was "almost perfect" (kappa = 0.81). Kendall's tau coefficient was 0.91, indicating high agreement. CONCLUSION: Sagittal STIR images detect facet joint edema in 14% of patients with low back pain. This fact may be useful for planning treatment including facet joint injections. PMID- 17410354 TI - Pseudoaneurysm of the superior gluteal artery following polytrauma. AB - Gluteal artery aneurysms are rare and often secondary to pelvic fractures, blunt or penetrating trauma. We describe a case of a superior gluteal artery pseudoaneurysm that presented as back pain with numbness and weakness of the lower extremities. Diagnosis was confirmed by color Doppler sonography and angiography. A proximal and distal control was obtained over the aneurysm neck via coil embolization with excellent hemostasis within the pseudoaneurysm and maintenance of perfusion to the left pelvis. PMID- 17410355 TI - High-grade surface osteosarcoma of the hand. AB - A 32-year-old woman presented with a 1-year history of mild pain in the right ring finger. Radiographs and CT revealed a calcified lesion with cortical erosion on the surface of the proximal aspect of the right ring finger proximal phalanx. On magnetic resonance imaging (MRI), the lesion showed low signal intensity on T1 and T2-weighted images and slight enhancement with gadolinium. Clinically, it was diagnosed as a benign bone-forming lesion such as florid reactive periostitis, and excision was accordingly performed. However, histological examination revealed proliferation of atypical osteoblastic cells among irregularly arranged osteoid seams. Taking the imaging findings into account, a pathological diagnosis of high-grade surface osteosarcoma was established. In general, bone- and cartilage-forming lesions of the hands and feet are benign. Osteosarcoma of short tubular bones in the hands and feet is extremely rare; moreover, high-grade surface osteosarcoma is one of the rarest subtypes of osteosarcoma. Nonetheless, high-grade surface osteosarcoma should be included in the differential diagnosis, particularly if the radiological findings or clinical course are not entirely typical of a more common benign process, to avoid incorrect clinicoradiological and pathological diagnosis. PMID- 17410357 TI - Anterior process calcaneal fractures: a systematic evaluation of associated conditions. AB - OBJECTIVE: The objective was to evaluate the association, by MRI, of anterior calcaneal process fractures with tarsal coalitions, ankle sprains, and bifurcate ligament abnormalities. DESIGN AND PATIENTS: A retrospective review of 1,479 foot and ankle MR images was performed, over a period of 5 years, for isolated anterior process fractures of the calcaneus. Fifteen 1.5-T MR examinations were systematically evaluated by two radiologists in consensus. Marrow edema patterns, presence of a calcaneonavicular coalition, as well as bifurcate and anterior talofibular ligaments, were evaluated. RESULTS: There were 15 fractures of the anterior calcaneal process with an incidence of 1%. The average patient age was 51 years (range 25-82). Twelve patients were women and 3 were men. The majority of the fractures (14 out of 15) presented as an edema pattern on T2-weighted images, either diffuse (9 out of 15), or vertical (5 out of 15). One case did not show marrow edema, but rather a hypointense line. Nine patients (60%) demonstrated calcaneonavicular coalition and anterior calcaneal process fracture. In 6 patients (50%) the anterior talofibular ligament (ATFL) was thickened. Three patients did not have axial images, and were classified as non-conclusive for the ATFL evaluation. The bifurcate ligament was thickened with hyperintense signal demonstrating a sprain in 9 out of 13 (69%). Only 2 patients (16.5%) had an anterior calcaneal process fracture without any associated abnormality. CONCLUSIONS: We believe that there is a probable association of anterior process fractures and calcaneonavicular coalitions. We also feel, based on our results and the prior literature that there is likely also an association with both ATFL injuries and bifurcate ligament injuries. PMID- 17410356 TI - The spine in Paget's disease. AB - Paget's disease (PD) is a chronic metabolically active bone disease, characterized by a disturbance in bone modelling and remodelling due to an increase in osteoblastic and osteoclastic activity. The vertebra is the second most commonly affected site. This article reviews the various spinal pathomechanisms and osseous dynamics involved in producing the varied imaging appearances and their clinical relevance. Advanced imaging of osseous, articular and bone marrow manifestations of PD in all the vertebral components are presented. Pagetic changes often result in clinical symptoms including back pain, spinal stenosis and neural dysfunction. Various pathological complications due to PD involvement result in these clinical symptoms. Recognition of the imaging manifestations of spinal PD and the potential complications that cause the clinical symptoms enables accurate assessment of patients prior to appropriate management. PMID- 17410358 TI - Imaging findings of avalanche victims. AB - OBJECTIVE: Skiing and hiking outside the boundaries remains an attractive wilderness activity despite the danger of avalanches. Avalanches occur on a relatively frequent basis and may be devastating. Musculoskeletal radiologists should be acquainted with these injuries. DESIGN AND PATIENTS: Fourteen avalanche victims (11 men and 3 women; age range 17-59 years, mean age 37.4 years) were air transported to a high-grade trauma centre over a period of 2 years. RESULTS: Radiographs, CT and MR images were prospectively evaluated by two observers in consensus. Musculoskeletal findings (61%) were more frequent than extraskeletal findings (39%). Fractures were most commonly seen (36.6%), involving the spine (14.6%) more frequently than the extremities (9.8%). Blunt abdominal and thoracic trauma were the most frequent extraskeletal findings. CONCLUSION: A wide spectrum of injuries can be found in avalanche victims, ranging from extremity fractures to massive polytrauma. Asphyxia remains the main cause of death along with hypoxic brain injury and hypothermia. PMID- 17410359 TI - NY-BR-1 protein expression in breast carcinoma: a mammary gland differentiation antigen as target for cancer immunotherapy. AB - NY-BR-1 is a recently identified differentiation antigen of the mammary gland. To use NY-BR-1 for T-cell-based immunotherapy, analysis of its co-expression with HLA class I antigens is required. In the present tissue microarray study, primary breast cancers (n = 1,444), recurrences (n = 88), lymph node (n = 525) and distant metastases (n = 91) were studied for NY-BR-1 expression using a novel monoclonal antibody. NY-BR-1 expression was compared with prognosis, estrogen receptor, HER2-status, EGFR and HLA class I antigen expression. NY-BR-1 was more frequently expressed in grade 1 (82%) than in grade 2 (69%) and grade 3 (46%) carcinomas (P < 0.0001). Moreover, NY-BR-1 expression correlated directly with estrogen receptor expression (P < 0.0001) and inversely correlated with HER2 status and EGFR expression (P < 0.0001 for both). Considering high expression level of co-expression, 198/1,321 (15%) primary breast carcinomas and 4/65 (6%) distant metastases expressed NY-BR-1 and HLA class I, suggesting that active immunotherapy can be applied to about 10% of breast cancer patients. Survival analysis showed an association of NY-BR-1 expression with better patient outcome (P = 0.015). No difference between NY-BR-1 expression of primary tumors and metastases could be found, indicating that the presence of NY-BR-1 in metastases can be deduced from their corresponding primary. Forty-three paired biopsies taken from patients before and after chemotherapy suggest that NY-BR-1 expression is not influenced by preceding chemotherapy (kappa = 0.89, P < 0.0001). In summary, the co-expression of NY-BR-1 with HLA class I antigens and its expression in metastases without modification by chemotherapy suggest that NY-BR 1 targeted immunotherapy represents a viable strategy in addition to other targeted cancer drug therapies of breast cancer. PMID- 17410360 TI - GM-CSF-secreting cancer immunotherapies: preclinical analysis of the mechanism of action. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF)-secreting tumor cell immunotherapies have demonstrated long-lasting, and specific anti-tumor immune responses in animal models. The studies reported here specifically evaluate two aspects of the immune response generated by such immunotherapies: the persistence of irradiated tumor cells at the immunization site, and the breadth of the immune response elicited to tumor associated antigens (TAA) derived from the immunotherapy. To further define the mechanism of GM-CSF-secreting cancer immunotherapies, immunohistochemistry studies were performed using the B16F10 melanoma tumor model. In contrast to previous reports, our data revealed that the irradiated tumor cells persisted and secreted high levels of GM-CSF at the injection site for more than 21 days. Furthermore, dense infiltrates of dendritic cells were observed only in mice treated with GM-CSF-secreting B16F10 cells, and not in mice treated with unmodified B16F10 cells with or without concurrent injection of rGM-CSF. In addition, histological studies also revealed enhanced neutrophil and CD4+ T cell infiltration, as well as the presence of apoptotic cells, at the injection site of mice treated with GM-CSF-secreting tumor cells. To evaluate the scope of the immune response generated by GM-CSF-secreting cancer immunotherapies, several related B16 melanoma tumor cell subclones that exist as a result of genetic drift in the original cell line were used to challenge mice previously immunized with GM-CSF-secreting B16F10 cells. These studies revealed that GM-CSF-secreting cancer immunotherapies elicit T cell responses that effectively control growth of related but antigenically distinct tumors. Taken together, these studies provide important new insights into the mechanism of action of this promising novel cancer immunotherapy. PMID- 17410361 TI - Treatment of non-small cell lung cancer patients with the trifunctional monoclonal antibody catumaxomab (anti-EpCAM x anti-CD3): a phase I study. AB - PURPOSE: Catumaxomab is a trifunctional monoclonal antibody with binding sites directed to human EpCAM and the human T cell antigen CD3 (anti-EpCAM x anti-CD3). Catumaxomab demonstrated efficacy when administered intraperitoneally in patients with EpCAM positive malignant ascites from ovarian cancer in terms of tumor cell killing and reduction of ascites generation. As EpCAM is also overexpressed in NSCLC, the present study was conducted in order to evaluate safety and tolerability of intravenous treatment with catumaxomab. PATIENTS AND METHODS: UICC stage IB-IV NSCLC patients were eligible, if they had at least one prior therapy. Other inclusion criteria were: age 18-75 years, adequate bone marrow function and AST or gamma-GT50% (++). Correlations between CXCR4 expression, data on patient and tumor features were studied by contingency tables and the chi2 test. Time-to-event curves were studied using the Kaplan Meier method. Univariate analysis was performed using the log-rank test. Ninety five percent confidence intervals (95% CI) of hazard ratios were also reported. RESULTS: Staining for CXCR4 protein was absent in 18 tumors (40.9%), present in <50% of cells in 19 (43.2%) and in >50% of cells in 7 (15.9%) tumors. CXCR4 expression correlated to the epithelioid-mixed cell type (P=0.030). No statistically significant relation emerged between CXCR4 expression, largest tumor diameter (LTD) and extracellular matrix patterns as evaluated through histological patterns stained with periodic acid-Schiff (PAS). Events occurred in 2 out of 18 patients (11.1%) with negative tumors (2 deaths), in 3 out of 19 patients (15.8%) with <50% of positive tumor cells (2 deaths and 1 occurrence of metastases) and in 1 out of 7 patients (14.3%) with >50% of positive tumor cells (1 occurrence of metastases). The cell type (P=0.0457) but not CXCR4 showed prognostic value at univariate analysis. CONCLUSION: This study shows that CXCR4 is commonly expressed in uveal melanoma and correlates with cell type a well established prognostic factor. PMID- 17410363 TI - Pedicle screw insertion: computed tomography versus fluoroscopic image guidance. AB - Computed tomography image-guided surgery (CTGS) clearly improves the accuracy of pedicle screw insertion. Recent reports claim that a fluoroscopy-guided system (FGS) offered high accuracy and easy application. However, the superiority of either technique remains unclear in clinical application. This study compares the accuracy of pedicle screws installed using CTGS with that of screws installed using FGS. Seventy-four screws inserted using FGS in 13 patients and 76 screws inserted using CTGS in 11 patients were compared. The study population included ten cases of vertebral fracture, five cases of degenerative spondylolisthesis, three cases of spondylolytic spondylolisthesis, two cases of tuberculous spondylitis, two cases of failed earlier back surgery and two case of ankylosing spondylitis with pseudarthrosis. The installed vertebral levels ranged from T8 to S1. Screw positions were assessed with postoperative radiographs and computed tomography. Sixty-nine (93.2%) screws were correctly placed in the FGS group, and seventy-three (96.1%) screws were correctly placed in the CTGS group (P=0.491). The results indicated that both image-guided systems offer high accuracy. However, the fluoroscope image-guided system could be considered the primary tool for lower thoracic and lumbosacral pedicle placement because it enables real-time navigation and does not require a preoperative CT scan. PMID- 17410364 TI - Intramedullary tibial nailing in distal third tibial fractures: distal locking screws and fracture non-union. AB - Distal third tibial fractures are prone to non-union following tibial nail insertion. The purpose of this study was to assess the union of distal third tibial fractures in patients who have undergone intra-medullary (IM) tibial nailing with one versus two distal locking screws. Sixty-five patients who had intramedullary tibial nail fixation were retrospectively analysed. Our results showed that 80% of non-unions in distal third fractures had only one distal locking screw compared to 20% who had two distal locking screws. This is statistically significant (p<0.01). We therefore conclude that two distal locking screws are essential for distal third fractures. PMID- 17410365 TI - Deltoid detachment consequent to open surgical repair of massive rotator cuff tears. AB - Deltoid detachment is one of the complications in open rotator cuff repair. Although it is often described, the actual prevalence, time at which it occurs and the predisposing causes are still unknown. We prospectively studied 112 patients with massive rotator cuff tears with a mean age of 67. The surgical approach was performed with a lateral para-acromial incision. Clinical assessment was performed with Constant's method. Of the 112 patients, 9 (8%) had deltoid detachment. It occurred about 3 months after surgery. Of the nine patients, two underwent revision surgery for the deltoid trans-bone reattachment. At the follow up, the patients with deltoid detachment had a mean increase of only 5.5 points in the Constant score compared to that of 16.9 obtained by the control group. Deltoid reattachment, performed on the two patients, provided a mean increase of 7 points only with respect to the post-operative control at the 4th month. Considering the unsatisfactory functional result consequent to deltoid detachment and the slight improvement obtained with the reattachment, we recommend the following: use suture thread thicker than #2, do not use a simple stitch and avoid extending acromioplasty to the lateral margin of the acromion. PMID- 17410366 TI - A case of complete heart block reverting to normal sinus rhythm after treatment for cardiac invasive Burkitt's lymphoma. PMID- 17410367 TI - Identification and characterization of genes involved in hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens x N. tabacum) using suppression subtractive hybridization. AB - Hybrid lethality is an important problem for cross-breeding; however, its molecular mechanism is not clear. The purpose of the present study was to identify the genes expressed during hybrid lethality in the hybrid cells (Nicotiana suaveolens x N. tabacum). In order to identify these genes, we employed suppression subtractive hybridization (SSH) between RNA isolated from cells expressing lethality (lethal hybrid line; LH line) and cells overcoming lethality fortuitously (a surviving hybrid line; SH line). Four populations of cDNA were created from the time points corresponding to before and during induction, and at and after the point of no return (PNR) during the process of programmed cell death (PCD) that occurs during hybrid lethality. By SSH and following dot-blot macroarray analysis, 99 genes out of 138 isolated clones were identified as hybrid lethality-related (HLR) genes. Quantitative real-time PCR analysis data indicated that ten clones were expressed specifically in LH line cells. The HLR genes in these clones show homology to genes involved in disease resistance, ethylene-induced reactions, phosphorylation, ubiquitination, jasmonic acid-related reactions, calcium signaling and self-incompatibility. These data suggested that at least some parts of the mechanism of hybrid lethality are shared with those of the putative functions of the HLR gene-related pathways. PMID- 17410368 TI - Acute appendicitis in a patient with hemolytic uremic syndrome: an unusual clinical scenario. AB - Gastroenteritis due to Escherichia coli O157:H7 occurs in young children and is associated with consumption of under cooked beef. Approximately 5-10% of patients will develop hemolytic uremic syndrome (HUS): renal failure, microangiopathic hemolytic anemia, and thrombocytopenia. A 6-year-old boy was admitted with abdominal pain, guaiac positive stool, decreased urine output and elevated creatinine levels. Hemodialysis was initiated upon rapid progression to anuria. On hospital day # 5 he developed acute abdominal pain, which was different from his initial assessment. Exam revealed focal tenderness in the right lower quadrant with localized guarding and rebound. Ultrasound demonstrated a dilated, fluid filled tubular structure in the RLQ concerning for appendicitis. Based on these findings the patient was taken to the operating room for a laparoscopic appendectomy. The patient had undergone dialysis the previous day and was preoperatively treated with DDAVP to minimize the risk of bleeding. The procedure occurred without complication and final pathology confirmed acute appendicitis. This case highlights the unique clinical scenario in which patients with HUS require operative intervention. Surgical procedures can be performed on these patients, however, all precautions should be taken to minimize the risk of bleeding, including the use of preoperative DDAVP. PMID- 17410369 TI - Influence of the neo-adjuvant radiochemotherapy as a factor in the surgical treatment of rectal cancer by expert surgeon. A comparative study. AB - BACKGROUND AND AIMS: Total mesorectal excision and surgeon experience are prognostic factors in rectal cancer surgery, in terms of local recurrence and conservative sphincter surgery. Pre-operative radiation-chemotherapy can even improve those results. The aim of this study is to assess the utility of pre operative radiation therapy (PRT) on the results of surgical treatment for rectal cancer comparing two consecutive series of patients operated on by surgeons with experience in rectal cancer surgery according to whether they had received PRT. MATERIALS AND METHODS: Retrospective review of 118 patients with rectal cancer, divided into two groups: group I, 57 patients without pre-operative radiation chemotherapy, and group II, 61 patients with rectal cancer who received pre operative radiation-chemotherapy. Both groups were homogeneous. The short-term results (surgical technique, post-operative stay, post-operative complications) and long-term results (local recurrences, liver metastases and both overall and tumour-related survival) were analysed. RESULTS: The rate of anterior resection in the lower third was significantly higher (p = 0.037) in group II than in group I (56 vs 23%), the rate of abdominoperineal resection in the middle third was significantly lower (p = 0.046) in group II (3.5 vs 21%). The incidence of post operative complications was similar in both groups, but the rate of anastomotic leaks was higher in group II. The local recurrences was lower in group II (p = 0.002), but the disease free survival and the overall survival was similar in both groups. At the multivariate analysis, the only prognostic factor for the local recurrence was the use of pre-operative radiation-chemotherapy. CONCLUSION: Besides surgeon experience and total mesorectal excision, a very important prognostic factor is the administration of pre-operative radiation-chemotherapy in cases of locally advanced rectal cancer, as it does not increase post operative morbidity and mortality and significantly influences the rate of local recurrences and the conservative sphincter surgery. PMID- 17410370 TI - Remodeling of the spine in spondylodiscitis of children at the age of 3 years or younger. AB - Spondylitis/spondylodiscitis is still an uncommon diagnosis often with a delay in diagnosis and treatment due to the uncharacteristic symptoms. The aim of this study is to increase the awareness and outline a pattern of investigation and treatment. We present six children with an average age of 23 months (19-33 months) at time of diagnosis, conservative treated and with a mean follow-up of 31 months (12-65 months). The evaluation included past medical history, clinical symptoms, X-rays, MRI-investigations and laboratory studies [CRP, erythrocytes sedimentation rate (ESR), white blood count (WBC) and blood cultures] during the course of treatment and follow-up. The predominate clinical findings were: limp, refusal to walk and/or back pain. The mean duration of symptoms until presentation at our clinic was 24 days (4-42 days), the final diagnosis was set after an average of 12 days (7-14 days). Laboratory findings were unspecific but ESR best correlated with the clinical symptoms during the therapy. Five patients were treated by parenteral antibiotics for a minimum of 3 weeks, followed by oral antibiotics adapted to the clinical and laboratory findings. One child received a combined antituberculous chemotherapy after positive skin test for tbc. All six children were immobilized with a body-plaster-cast for an average time of 15 weeks (5-26 weeks). Four patients additionally were treated by further corset therapy for an average of 10 months (3-18 months). Radiological findings on plane X-rays (a.p. and lateral views) at time of diagnosis were decreased height of the disk space and erosions of adjacent vertebral endplates and residues of these radiological changes with signs of bony healing (sclerotic vertebral endplates or partial fusion) were seen at the latest follow-up. There was no case of instability or deformity like scoliosis or kyphosis. The MRI showed the earliest detectable typical vertebral bone involvements and confirmed the diagnosis in combination with laboratory findings and clinical symptoms. Spondylitis/spondylodiscitis should be considered as diagnosis in children with refusal to walk or gait disturbances especially in combination with elevated ESR. MRI is the tool of choice to set the diagnosis early. With an adequate and early therapy of bracing (body-plaster-cast), antibiotics and clinical monitoring good long-term result without spine instability or deformity can be achieved. PMID- 17410371 TI - Open grade III "floating ankle" injuries: a report of eight cases with review of literature. AB - INTRODUCTION: "Floating ankle" injuries result from high-energy trauma and are usually associated with extensive soft tissue damage. Such rare and complex injuries in the acute phase pose therapeutic dilemma to the treating surgeon. When salvage instead of amputation is preferred, a variety of treatment options are available ranging from open reduction along with minimal internal fixation to external fixation. In this retrospective case series study we report eight patients with open "floating ankle" injuries who had been treated with external fixation with or without internal fixation. MATERIALS AND METHODS: Eight consecutive patients (7M: 1F) with mean age of 28 years (range 18-35 years) were treated. All had Grade III open fractures of the distal tibia with foot fractures and various degrees of soft tissue injuries. ISS and MESS were used to assess injury severity. Immediate radical wound debridement; skeletal stabilization and early soft tissue coverage were done by combined trauma and plastic surgical services followed by standard post-operative rehabilitation. All the patients were assessed with SF-36 questionnaire at mean follow-up of 2.5 years (range 2-4 years). RESULTS: Three patients were treated primarily with Ilizarov ring fixators, one had hybrid fixation and the other four had Hoffman frames. Four patients needed further surgeries. One patient developed metatarsal osteomyelitis, which was debrided and treated with antibiotics. The SF-36 results were compared with age matched UK norms for men and women in all categories. Only two patients returned to their previous employment. Six suffered varying degrees of disability out of which one underwent amputation. CONCLUSIONS: External fixation with or without internal fixation is an option when salvaging rare injuries like open grade III "floating ankle" injuries. Salvaged patients do suffer a cocktail of crippling disease characterized by psycho-socio-economic and physical disability and result in increased hospital costs. We recommend more prospective studies with long-term follow-up of such complex injuries to identify the indications for salvage and also the criteria for a successful salvage. PMID- 17410372 TI - Histopathologic findings in women with postmenopausal bleeding: implication for endometrial thickness and circulating levels of sex steroid hormones. AB - OBJECTIVE: The aim of this study is to compare the relationship between estrone (E1), estradiol (E2), androgens, and prolactin blood levels on the one hand, and endometrial thickness and related histopathologic results on the other, in postmenopausal women admitted with uterine bleeding. METHODS: The study was conducted in Gazi University School of Medicine, Obstetrics and Gynecology Clinic with a total of 128 patients. The study group consisted of 64 postmenopausal patients admitted with uterine bleeding, whereas the control group consisted of 64 healthy postmenopausal women. Vaginal sonography was performed to evaluate the endometrial thicknesses of the patients. Serum levels of free testosterone, androstenedione and estrone (E1) were determined by radioimmunoassay while serum estradiol (E2), prolactin, and dehydroepiandrosterone sulfate (DHEA-S) levels were evaluated by chemiluminescent method. RESULTS: The median age, duration of menopause, menopausal age and gravidity, and parity did not differ between women with postmenopausal bleeding and the control group (P > 0.05). However, DHEA-S level was lower (P < 0.05) and endometrial thickness was greater in the study group than the control group (P < 0.05). Furthermore, the study identified that median endometrial thickness of the patients in atrophic endometrium group was less than the endometrial hyperplasia and endometrium carcinoma group (P < 0.05). CONCLUSION: In the current study, all these hormones seemed to be indifferent between groups of endometrial cancer and other pathological results. Based on our results regarding the safe margin of endometrial thickness in women with postmenopausal bleeding, it seems justifiable to refrain from curettage in patients with an endometrium of < or =4 mm. PMID- 17410373 TI - Outcomes of the first midwife-led birth centre in Italy: 5 years' experience. AB - OBJECTIVE: To assess the experience of the first 5 years of the first midwife-led birth centre (MLBC) in Italy. STUDY DESIGN: Data were prospectively collected to analyse the first 5 years' experience of the MLBC. MLBC is located alongside a University hospital maternity unit and it offers care to women with a straightforward pregnancy and midwives take primary professional responsibility for care. Women with maternal diseases, complicated obstetric history, height < 150 cm, maternal age > 45, or multiple pregnancy were excluded. Transfer was request in case of antenatal, intrapartum and postpartum pathological conditions. RESULTS: During the 5-year period (1 January 2001-31 December 2005), 1,438 low risk women were admitted in labour to the MLBC. Of these, 203 (14.1%) were transferred during labour to consultant care (138 because of pathologies and 65 because of request of epidural analgesia). Among the transfers, the caesarean sections were 87, corresponding to 6.1% (87/1,438) of the total of women admitted to MLBC, while the operative vaginal deliveries were 14, corresponding to 1.0% (14/1,438) of the total of women admitted to MLBC. Among women who gave birth in the MLBC, episiotomy rate was 17.1%. CONCLUSIONS: In Italy, in the passed 10 years, the caesarean section rate reached 60%, in some regions. According to our data, the first 5 years of activity of the first MLBC in Italy had been associated with a low rate of medical interventions during labour and birth, with high rates of spontaneous vaginal birth and without signs of complications. We hope that this experience could be taken as a model to improve the quality of maternity care in Italy. PMID- 17410374 TI - Self-reported symptoms and their effects on cognitive functioning in workers with past exposure to solvent-based glues: an 18-year follow-up. AB - OBJECTIVES: To examine to what extent exposure to organic solvents during the working life affects general well-being in the long term, and to explore the relationship between self-reported symptoms and cognitive functioning in previously solvent-exposed floor layers. METHODS: The study included 41 solvent exposed floor layers and 40 unexposed referents participating in a longitudinal follow-up study 18 years after the baseline assessment. Symptom prevalence and level of spare time activities were studied using the same methods as in the initial study. These include a general health examination, the Q16 symptom questionnaire, and a questionnaire for spare time activities. Relationships between symptoms and cognitive functioning were analysed based on recently published data on cognitive functioning of the participants at follow-up. RESULTS: At follow-up neuropsychiatric symptoms such as need to check things, depressive mood, and abnormal fatigue, were more prevalent among floor layers, particularly the most exposed individuals, than among referents. In addition, the most highly exposed floor layers reported more concentration difficulties and irritability. Fatigue and depressive mood increased over the follow-up time in the most exposed floor layers but not in the referents. Memory difficulties, although more frequent among floor layers than among referents, had decreased in floor layers while increased in referents. Floor layers also reported some negative effects on intimate relations and activity level. Neuropsychiatric symptoms were related to poorer performance chiefly in memory tasks and tests of complex attention and perceptual speed, more seldom in visuospatial tasks. CONCLUSIONS: Findings of exposure-related, long-lasting, partly deteriorating neuropsychiatric complaints indicate that general well-being later in life has been affected in floor layers with past heavy solvent exposure. We also found frequent associations between symptom prevalence and the cognitive functioning. Together with previous findings of dose-related cognitive decrements, the present results strengthen the evidence that long-term heavy occupational solvent exposure may negatively interact with the normal ageing process. PMID- 17410375 TI - Hand vibration: non-contact measurement of local transmissibility. AB - OBJECTIVES: Grip and push forces required for the use of vibrating tools are considered important influencing inputs for the assessment of hand-vibration transmissibility (TR). At present TR measurements are usually referred to the palm of the hand The aims of the present paper are: to present an original measurement procedure for non-contact assessment of the transmissibility of fingers; to report TR data measured on six points of the hand of nine subjects; to correlate TR with: grip, push, hand volume and BMI. METHODS: Tests have been carried out using a cylindrical handle mounted on an shaker. A laser Doppler vibrometer is used to measure the vibration velocity. Push force is measured using a force platform, whereas grip force is measured using a capacitive pressure sensor matrix wrapped around the handle. Tests have been conducted on nine healthy subjects. Proximal and distal regions of the second, fourth and fifth fingers have been investigated. Tests were carried out using a push force of: 25, 50 and 75 N. The excitation signal was a broadband random vibration in the band 16-400 Hz with un-weighted rms acceleration level of 6 m/s(2). RESULTS: Results show how in general TR values measured on distal points are higher respect to the proximal points. A resonance peak is present for all the measured points in the band 55-80 Hz. ANOVA analysis showed that TR is not significantly dependent on: BMI, hand volume and push force alone. While TR is significantly dependent on: grip force alone, measurement positions and grip and push force together. CONCLUSIONS: The proposed procedure shows the advantage to allow local vibration measurement directly on the fingers without the necessity to apply any contact sensor. Results demonstrate how the transmissibility is significantly different on the point where the acceleration is measured. PMID- 17410376 TI - Work-related physical and psychosocial risk factors for sick leave in patients with neck or upper extremity complaints. AB - OBJECTIVES: To study work-related physical and psychosocial risk factors for sick leave among patients who have visited their general practitioner for neck or upper extremity complaints. METHODS: Three hundred and forty two patients with neck or upper extremity complaints completed self-report questionnaires at baseline and after 3 months. Cox regression models were used to investigate the association between work-related risk factors and sick leave (i.e., lost days from work due to neck or upper extremity complaints in 3 months). Effect modification by sick leave at baseline, sex, worrying and musculoskeletal co morbidity was evaluated by adding product terms to the regression models. RESULTS: In the subgroup of patients who scored high on the pain copying scale "worrying" the hazard ratio of sick leave was 1.32 (95% CI 1.07-1.62) per 10% increase in heavy physical work. The subgroup of patients who were sitting for long periods of time had a reduced risk of sick leave as compared to patients who did not spend a lot of time sitting, again only in patients who scored high on the pain coping scale "worrying" (adjusted HR=0.17, 95%-CI 0.04-0.72). Other work related risk factors were not significantly related to sick leave. CONCLUSIONS: Heavy physical work increased the risk of sick leave and prolonged sitting reduced the risk of sick leave in a subgroup of patients who worried much about their pain. Additional large longitudinal studies of sufficiently large size among employees with neck or upper extremity complaints are needed to confirm our results. PMID- 17410377 TI - Neuronal selectivity, population sparseness, and ergodicity in the inferior temporal visual cortex. AB - The sparseness of the encoding of stimuli by single neurons and by populations of neurons is fundamental to understanding the efficiency and capacity of representations in the brain, and was addressed as follows. The selectivity and sparseness of firing to visual stimuli of single neurons in the primate inferior temporal visual cortex were measured to a set of 20 visual stimuli including objects and faces in macaques performing a visual fixation task. Neurons were analysed with significantly different responses to the stimuli. The firing rate distribution of 36% of the neurons was exponential. Twenty-nine percent of the neurons had too few low rates to be fitted by an exponential distribution, and were fitted by a gamma distribution. Interestingly, the raw firing rate distribution taken across all neurons fitted an exponential distribution very closely. The sparseness a (s) or selectivity of the representation of the set of 20 stimuli provided by each of these neurons (which takes a maximal value of 1.0) had an average across all neurons of 0.77, indicating a rather distributed representation. The sparseness of the representation of a given stimulus by the whole population of neurons, the population sparseness a (p), also had an average value of 0.77. The similarity of the average single neuron selectivity a (s) and population sparseness for any one stimulus taken at any one time a (p) shows that the representation is weakly ergodic. For this to occur, the different neurons must have uncorrelated tuning profiles to the set of stimuli. PMID- 17410378 TI - A membrane-associated NAC transcription factor regulates salt-responsive flowering via FLOWERING LOCUS T in Arabidopsis. AB - The plant-specific NAC (NAM, ATAF1/2, CUC2) transcription factors play regulatory roles in diverse developmental processes and stress responses. Notably, a subset of the NAC members, collectively designated NTLs, is membrane-associated, suggesting that membrane-mediated regulation would be an important molecular scheme functioning in rapid transcriptional responses to external stimuli. Here, we examined the physiological role of a salt-responsive NTL, NTL8, by molecular genetic and transgenic approaches. NTL8 is expressed as a membrane-associated, dormant form and subsequently processed into a transcriptionally active, nuclear form. Transgenic plants overproducing an active NTL8 form exhibited delayed flowering as well as reduced growth with small curled leaves. Accordingly, expression of FLOWERING LOCUS T (FT) and its downstream genes was significantly reduced in the transgenic plants. Furthermore, FT was significantly repressed by high salt. We therefore propose that NTL8 mediates salt-responsive flowering via FT in Arabidopsis. PMID- 17410379 TI - Affective stimulus properties influence size perception and the Ebbinghaus illusion. AB - In the New Look literature of the 1950s, it has been suggested that size judgments are dependent on the affective content of stimuli. This suggestion, however, has been 'discredited' due to contradictory findings and methodological problems. In the present study, we revisited this forgotten issue in two experiments. The first experiment investigated the influence of affective content on size perception by examining judgments of the size of target circles with and without affectively loaded (i.e., positive, neutral, and negative) pictures. Circles with a picture were estimated to be smaller than circles without a picture, and circles with a negative picture were estimated to be larger than circles with a positive or a neutral picture confirming the suggestion from the 1950s that size perception is influenced by affective content, an effect notably confined to negatively loaded stimuli. In a second experiment, we examined whether affective content influenced the Ebbinghaus illusion. Participants judged the size of a target circle whereby target and flanker circles differed in affective loading. The results replicated the first experiment. Additionally, the Ebbinghaus illusion was shown to be weakest for a negatively loaded target with positively loaded and blank flankers. A plausible explanation for both sets of experimental findings is that negatively loaded stimuli are more attention demanding than positively loaded or neutral stimuli. PMID- 17410380 TI - Live and let die: manipulation of host hepatocytes by exoerythrocytic Plasmodium parasites. AB - The generation of rodent Plasmodium strains expressing fluorescent proteins in all life cycle stages has had a big impact on malaria research. With this tool in hand, for the first time it was possible to follow in real time by in vivo microscopy the infection route of Plasmodium sporozoites transmitted to the mammalian host by Anopheles mosquitoes. Recently, this work has been extended to the analysis of both hepatocyte infection by Plasmodium sporozoites, as well as liver merozoite transport into blood vessels. The stunning results of these studies have considerably changed our understanding of hepatocyte invasion and parasite liberation. Here, we describe the most important findings of the last years and in addition, we elaborate on the molecular events during the intracellular development of Plasmodium exoerythrocytic forms that give rise to erythrocyte infecting merozoites. PMID- 17410381 TI - Malleable skin coloration in cephalopods: selective reflectance, transmission and absorbance of light by chromatophores and iridophores. AB - Nature's best-known example of colorful, changeable, and diverse skin patterning is found in cephalopods. Color and pattern changes in squid skin are mediated by the action of thousands of pigmented chromatophore organs in combination with subjacent light-reflecting iridophore cells. Chromatophores (brown, red, yellow pigment) are innervated directly by the brain and can quickly expand and retract over underlying iridophore cells (red, orange, yellow, green, blue iridescence). Here, we present the first spectral account of the colors that are produced by the interaction between chromatophores and iridophores in squid (Loligo pealeii). Using a spectrometer, we have acquired highly focused reflectance measurements of chromatophores, iridophores, and the quality and quantity of light reflected when both interact. Results indicate that the light reflected from iridophores can be filtered by the chromatophores, enhancing their appearance. We have also measured polarization aspects of iridophores and chromatophores and show that, whereas structurally reflecting iridophores polarize light at certain angles, pigmentary chromatophores do not. We have further measured the reflectance change that iridophores undergo during physiological activity, from "off" to various degrees of "on", revealing specifically the way that colors shift from the longer end (infra-red and red) to the shorter (blue) end of the spectrum. By demonstrating that three color classes of pigments, combined with a single type of reflective cell, produce colors that envelop the whole of the visible spectrum, this study provides an insight into the optical mechanisms employed by the elaborate skin of cephalopods to give the extreme diversity that enables their dynamic camouflage and signaling. PMID- 17410382 TI - Persistent pain in patients following scoliosis surgery. AB - Chronic or persistent pain is increasingly recognised as a consequence of surgery in a number of different disciplines. The pain often exhibit qualities that differ from the acute post-operative pain and may represent changes in the central nervous system. There is lack of information regarding the incidence of persistent pain in patients following spinal surgery for scoliosis. This study aims to estimate the incidence of persistent pain following spinal surgery for scoliosis in a group of mainly adolescent patients. Questionnaires were distributed to consecutive patients attending the outpatient clinic of a hospital with specialist services in paediatric orthopaedics and spinal surgery. One hundred and five patients out of 122 eligible patients completed the survey. Fifty-two percent had ongoing pain upon hospital discharge either in the primary surgical site and/or in the iliac bone graft site. Approximately 10 and 7% of all patients had back and pelvic pain persisting beyond 12 months, respectively. A small proportion described elements of neuropathic pain. There was a trend suggesting that those who experienced more severe post-operative pain were more likely to develop persistent pain. These data are consistent with those reports that implicate surgery as the trigger for chronic pain. PMID- 17410383 TI - The anatomical relationship between the aorta and the thoracic vertebral bodies and its importance in the placement of the screw in thoracoscopic correction of scoliosis. AB - Thoracoscopically-assisted anterior spinal instrumentation is being used widely to treat adolescent idiopathic scoliosis (AIS). Recent studies have showed that screws placed thoracoscopically could counter the aorta or entrance into the spinal canal. There are a few studies defining the anatomic landmarks to identify the relationship between the aorta and the thoracic vertebral body using quantitative measurement for the sake of safe placement of thoracoscopic vertebral screw in anterior correction for AIS. The CT scanning from T4 to T12 in 64 control subjects and 30 AIS patients from mainland China were analyzed manually. Parameters to be measured included the angle for safety screw placement (alpha), the angle of the aorta relative to the vertebral body (beta), the distance from the line between the left and the right rib heads to the anterior wall of the vertebral canal (a), the distance from the left rib head to posterior wall of the aorta (b), the vertebral body transverse diameter (c) and vertebral rotation (gamma). No significant differences were found between the groups with respect to age or sex. Compared with the control group, alpha angle from T7 to T10, beta angle from T5 to T10 and b value at T9, T10 were significantly lower in the scoliotic group. The a value was significantly lower in the scoliotic group. The c value showed no significant difference between the two groups. In conclusion, to place the thoracoscopic vertebral screw safely, at the cephalad thoracic spine (T4-T6), the maximum ventral excursion angle should decrease gradually from 20 degrees to 5 degrees , the entry-point of the screw should be close to the rib head. For apical vertebrae (T7-T9), the maximum ventral excursion angle increased gradually from 5 degrees to 12 degrees. At the caudal thoracic spine (T10-T12), the maximum ventral excursion angle increased, the entry-point should shift 3 approximately 5 mm ventrally. PMID- 17410384 TI - Evidence for heme release in layer-by-layer assemblies of myoglobin and polystyrenesulfonate on pyrolitic graphite. AB - Layer-by-layer assemblies of myoglobin and polystyrenesulfonate (PSS) on pyrolitic graphite have been investigated with the goal of determining the origin of the voltammetric response of these films. From the similar midpoint potential, coverage and electron transfer behavior compared with those of adsorbed free heme, it was concluded that the observed voltammetric peak is due to heme adsorbed at the electrode surface. This suggests that the interactions between the pyrolitic graphite electrode, PSS and myoglobin can result in heme release from the protein followed by heme adsorption on the electrode. PMID- 17410385 TI - The prevalence of myofascial pain and its association with occlusal factors in a threshold country non-patient population. AB - The objective of the study is to assess the prevalence of myofascial pain in a threshold country and to isolate occlusal risk factors. One hundred and seventy one randomized selected women were examined by a trained examiner in accordance with the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) examination procedure. Subscales of the SCL 90-R, graded chronic pain status, and anamnestic questionnaires were also used. Logistic regression was performed to compute the odds ratios for six common occlusal features with regard to the presence of myofascial pain, in accordance with the RDC/TMD criteria. Fifteen subjects (15 / 151 = 9.93%) suffered from myofascial pain. Results from logistic regression analysis showed that non-occlusion (posterior teeth, at least one side) and open bite increased the risk of myofascial pain. The prevalence of myofascial pain in this study is comparable with that in another study, in a highly industrialized environment, in which the RDC/TMD was used. The role of occlusion in a non-patient population seems to be restricted to serious alterations of normality. This article presents the prevalence of myofascial pain and its association with occlusal factors. This issue will help the clinicians to assess the influence of occlusion in myofascial pain patients and to send the patient to the appropriate specialist. PMID- 17410386 TI - Efficacy and tolerability of prolonged linezolid therapy in the treatment of orthopedic implant infections. AB - The aim of the study presented here was to assess the efficacy and tolerability of linezolid in the treatment of orthopedic implant infections (OII). Eighty-five patients with an OII treated with linezolid were prospectively followed up for a minimum of 12 months from the end of antibiotic therapy. Outcome was evaluated in relation to the duration and type of symptoms (acute or chronic) and the retention or removal of the implant. For acute and chronic infections, the respective success rates were 100 and 92.3% when the implant was removed and 72.2 and 42.8% when it was not. The median length of linezolid treatment in acute and chronic infections was 47 and 60 days, respectively. Thrombocytopenia was observed in four (4.7%) patients and anemia in five (5.8%). The results suggest oral linezolid is an effective and well-tolerated alternative for treating OII. PMID- 17410387 TI - In vitro susceptibility of methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium to daptomycin and other antibiotics. PMID- 17410388 TI - Cytomegalovirus DNA in semen of men seeking fertility evaluation in Abidjan, Cote d'Ivoire. PMID- 17410389 TI - MDCT coronary angiography for diagnosis of anomalous origin right coronary artery: a case report. AB - This is a case report of anomalous origin of the right coronary artery, which arises from the left sinus of Valsalva depicted with multi-detector row tomography (MDCT) coronary angiography performed in emergency. In symptomatic patient, this anomalous origin is considered a malignant anomaly and needs urgent surgical treatment. Some studies demonstrated that MDCT is a reliable non invasive technique to identify and define anomalous coronary arteries (ACAs) and their course. PMID- 17410390 TI - Midgut volvulus in an adult patient with malrotation and abdominal heterotaxia: a case report. AB - Midgut volvulus is rare in adulthood and if not diagnosed accurately, carries a high mortality rate. We present a case of a young adult man who presented with acute abdominal pain and was found to have malrotation with abdominal heterotaxia. Abdominal computed tomography can identify anomalies associated with intestinal malrotation. With the awareness of the potential for malrotation to predispose to midgut volvulus, the patient and his physician will have a higher index of suspicion when abdominal pain occurs. Appropriate treatment of midgut volvulus will reduce morbidity and mortality. PMID- 17410391 TI - In vivo quantification of response to treatment in patients with multiple myeloma by 1H magnetic resonance spectroscopy of bone marrow. AB - OBJECT: Magnetic resonance imaging (MRI) is the gold standard non-invasive technique to detect malignant disease in the bone marrow. Proton magnetic resonance spectroscopy (MRS) can be performed as a quick adjunct to routine spinal MRI. We performed proton MRS to patients with multiple myeloma (MM) at diagnosis and after treatment to investigate the possible correlation of MRS data with response to therapy. PATIENTS AND METHODS: Twenty-one patients with newly diagnosed MM underwent combined MRI/MRS explorations of a transverse center section in the fifth lumbar vertebral body. MRS was acquired with STEAM and 40 ms TE. Areas of unsuppressed water and lipid resonances were used to calculate the lipid-to-water ratio (LWR). RESULTS: No association was detected between initial LWRs and the clinical characteristics of patients. Post treatment MRS was available in 16 patients of whom 11 (69%) presented an LWR increase, this included all complete responders (8/8, 100%, P = 0.012). A post-treatment LWR value equal to or larger than one is proposed as a non-invasive marker of complete response to treatment. CONCLUSION: Only patients responding to treatment presented a significant increase in bone marrow LWR after therapy. MRS may provide an adequate quantification of response to chemotherapy in patients with MM. PMID- 17410392 TI - The fine art of CPAP titration--will it ever become obsolete? PMID- 17410393 TI - [Cranial nerve deficits caused by uncommon skull base lesions of the cavernous sinus]. PMID- 17410394 TI - A phylogenetic evaluation of whether endophytes become saprotrophs at host senescence. AB - Fungal endophytes and saprotrophs generally play an important ecological role within plant tissues and dead plant material. Several reports based solely on morphological observations have postulated that there is an intimate link between endophytes and saprotrophs. This study aims to provide valuable insight as to whether some endophytic fungi manifest themselves as saprotrophs upon host decay. Ribosomal DNA-based sequence comparison and phylogenetic relationships from 99 fungal isolates (endophytes, mycelia sterilia, and saprotrophs) recovered from leaves and twigs of Magnolia liliifera were investigated in this study. Molecular data suggest there are fungal taxa that possibly exist as endophytes and saprotrophs. Isolates of Colletotrichum, Fusarium, Guignardia, and Phomopsis, which are common plant endophytes, have high sequence similarity and are phylogenetically related to their saprotrophic counterparts. This provides evidence to suggest that some endophytic species change their ecological strategies and adopt a saprotrophic lifestyle. The implication of these findings on fungal biodiversity and host specificity is also discussed. PMID- 17410395 TI - Ecological variables affecting predatory success in Myxococcus xanthus. AB - The feeding efficiency of microbial predators depends on both the availability of various prey species and abiotic variables. Myxococcus xanthus is a bacterial predator that searches for microbial prey by gliding motility, and then kills and lyses its prey with secreted compounds. We manipulated three ecological variables to examine their effects on the predatory performance of M. xanthus to better understand its behavior and how it affects prey populations. Experiments were designed to determine how surface solidity (hard vs soft agar), density of prey patches (1 vs 2 cm grids), and type of prey (Gram-positive Micrococcus luteus vs Gram-negative Escherichia coli) affect predatory swarming and prey killing by M. xanthus. The prey were dispersed in patches on a buffered agar surface. M. xanthus swarms attacked a greater proportion of prey patches when patches were densely arranged on a hard-agar surface, compared with either soft-agar surfaces or low-patch-density arrangements. These ecological variables did not significantly influence the rate of killing of individual prey within a patch, although a few surviving prey were more likely to be recovered on soft agar than on hard agar. These results indicate that M. xanthus quickly kills most nearby E. coli or M. luteus regardless of the surface. However, the ability of M. xanthus to search out patches of these prey is affected by surface hardness, the density of prey patches, and the prey species. PMID- 17410396 TI - Phenotypically different microalgal morphospecies with identical ribosomal DNA: a case of rapid adaptive evolution? AB - The agents driving the divergence and speciation of free-living microbial populations are still largely unknown. We investigated the dinoflagellate morphospecies Scrippsiella hangoei and Peridinium aciculiferum, which abound in the Baltic Sea and in northern temperate lakes, respectively. Electron microscopy analyses showed significant interspecific differences in the external cellular morphology, but a similar plate pattern in the characteristic dinoflagellate armor. Experimentally, S. hangoei grew in a wide range of salinities (0-30), whereas P. aciculiferum only grew in low salinities (0-3). Despite these phenotypic differences and the habitat segregation, molecular analyses showed identical ribosomal DNA sequences (ITS1, ITS2, 5.8S, SSU, and partial LSU) for both morphospecies. Yet, a strong interspecific genetic isolation was indicated by amplified fragment length polymorphism (F (ST) = 0.76) and cytochrome b (cob) sequence divergence ( approximately 1.90%). Phylogenetic reconstructions based on ribosomal (SSU, LSU) and mitochondrial (cob) DNA indicated a recent marine ancestor for P. aciculiferum. In conclusion, we suggest that the lacustrine P. aciculiferum and the marine-brackish S. hangoei diverged very recently, after a marine-freshwater transition that exposed the ancestral populations to different selective pressures. This hypothetical scenario agrees with mounting data indicating a significant role of natural selection in the divergence of free living microbes, despite their virtually unrestricted dispersal capabilities. Finally, our results indicate that identical ITS rDNA sequences do not necessarily imply the same microbial species, as commonly assumed. PMID- 17410397 TI - Subintimal double-barrel restenting of an occluded primary stented superficial femoral artery. AB - In-stent re-stenosis is a frequent complication of endovascular stents, especially in the superficial femoral artery (SFA). Endovascular re-intervention of in- or peri-stent occlusive disease consists of recanilization through the occluded stent. In our case report, we describe the endovascular treatment of a previously placed stent in the SFA. We unintentionally passed the affected stent subintimally, in a double barrel fashion next to the first stent. The procedure was without any complications and with a successfull angiographic result. At one year follow-up the patient still has no complaints and the stent is still patent. PMID- 17410399 TI - Guidelines for training in diagnostic and therapeutic endoscopic retrograde cholangiopancreatography (ERCP). PMID- 17410400 TI - Guidelines for deep venous thrombosis prophylaxis during laparoscopic surgery. PMID- 17410401 TI - Medical liability insurance claims on entry-related complications in laparoscopy. AB - BACKGROUND: Installation of the pneumoperitoneum is an essential part of laparoscopic surgery. Creation can be performed by either the open or a closed technique. The aim of this study was to assess the number of and contributing factors to entry-related complications in medical liability insurance claims in The Netherlands. METHODS: A retrospective chart review was performed, including all malpractice claims filed at MediRisk, which is presently the largest medical liability mutual insurance company for institutions, mainly hospitals, in healthcare in The Netherlands. RESULTS: From January 1993 to December 2005, 41 claims were identified as entry-related complications which comprised 18% of all laparoscopy-related complications leading to claims. Most were young (median age = 35 years) female patients who had routine, nonadvanced, laparoscopic procedures planned as short-stay or day-care procedures. The claims were equally divided between general surgery (n = 20) and gynecology (n = 21). A total of 51 structures were injured. There were 18 vascular structure injuries, 30 bowel injuries, and three other injuries. An open entry technique was used in only two (5%) patients. Vascular injury was exclusively associated with closed entry. In only 19 (46%) patients the entry-related complication was diagnosed peroperatively, consisting of 70% of the vascular and 25% of the bowel injuries. Twenty-six patients (64%) were admitted to the intensive care unit for a median of five days. There was no mortality. Besides conversion, the majority of the patients filed a claim to compensate for a longer hospital stay and related costs. A payment was made in 17 (57%) of the 30 settled claims. CONCLUSIONS: Medical liability claims concerning laparoscopic entry-related complications comprised a fifth of all laparoscopy-related claims. Claims concerning entry related complications occurred in young patients who had routine, nonadvanced procedures. In the investigated cases most claims involved the closed-entry technique. PMID- 17410402 TI - Factors associated with frequency of emergency department visits for chronic obstructive pulmonary disease exacerbation. AB - BACKGROUND: Little is known about the factors associated with frequency of emergency department visits (FEDV) in chronic obstructive pulmonary disease (COPD) patients with recurrent exacerbations. OBJECTIVE: To characterize the use of emergency department (ED) services in patients with COPD exacerbation and identify factors associated with FEDV. DESIGN: A prospective, multicenter cohort study. PATIENTS: Three hundred eighty-eight patients were included. Fifty-two percent were women and the median age was 69 years (interquartile range 62-76). MEASUREMENTS: Using a standard questionnaire, consecutive ED patients with COPD exacerbation were interviewed. The number of ED visits in the previous year was retrospectively collected. RESULTS: Over the past year, this cohort reported a total of 1,090 ED visits because of COPD exacerbation. Thirteen percent of COPD patients had 6 or more ED visits, accounting for 57% of the total ED visits in the past year. Multivariate analysis showed that patients with an increased FEDV were more likely to be Hispanic (incidence rate ratio [IRR] 1.97, 95% confidence interval [CI] 1.16-3.33), to have more severe COPD as determined by previous hospitalizations (IRR 2.06, 95% CI 1.51-2.82), prior intubations (IRR 1.49, 95% CI 1.02-2.18), prior use of systemic corticosteroids (IRR 1.57, 95% CI 1.16-2.13) and methylxanthine (IRR 1.48, 95% CI 1.04-2.12), and less likely to have a primary care provider (IRR 0.51, 95% CI 0.31-0.82). CONCLUSIONS: Our results suggest that both disease and health care-related factors were associated with FEDV in COPD exacerbation. Multidisciplinary efforts through primary care provider follow-up should be assessed to test the effects on reducing the high morbidity and cost of recurrent COPD exacerbations. PMID- 17410403 TI - Patients at-risk for cost-related medication nonadherence: a review of the literature. AB - OBJECTIVE: Up to 32% of older patients take less medication than prescribed to avoid costs, yet a comprehensive assessment of risk factors for cost-related nonadherence (CRN) is not available. This review examined the empirical literature to identify patient-, medication-, and provider-level factors that influence the relationship between medication adherence and medication costs. DESIGN: We conducted searches of four databases (MEDLINE, CINAHL, Sciences Citations Index Expanded, and EconLit) from 2001 to 2006 for English-language original studies. Articles were selected if the study included an explicit measure of CRN and reported results on covarying characteristics. MAIN RESULTS: We found 19 studies with empirical support for concluding that certain patients may be susceptible to CRN: research has established consistent links between medication nonadherence due to costs and financial burden, but also to symptoms of depression and heavy disease burden. Only a handful of studies with limited statistical methods provided evidence on whether patients understand the health risks of CRN or to what extent clinicians influence patients to keep taking medications when faced with cost pressures. No relationship emerged between CRN and polypharmacy. CONCLUSION: Efforts to reduce cost-related medication nonadherence would benefit from greater study of factors besides the presence of prescription drug coverage. Older patients with chronic diseases and mood disorders are at-risk for CRN even if enrolled in Medicare's new drug benefit. PMID- 17410404 TI - Optimization of dose and image quality for computed radiography and digital radiography. PMID- 17410406 TI - Relationship between (Na + K)-ATPase activity, lipid peroxidation and fatty acid profile in erythrocytes of hypertensive and normotensive subjects. AB - Oxidative stress may play a role in the pathogenic mechanism of essential hypertension. Lipid peroxidation can alter the cellular structure of membrane bound enzymes by changing the membrane phospholipids fatty acids composition. We investigated the relationship between (Na + K)-ATPase activity, lipid peroxidation, and erythrocyte fatty acid composition in essential hypertension. The study included 40 essential hypertensive and 49 healthy normotensive men (ages 35-60 years). Exclusion criteria were obesity, dyslipidemia, diabetes mellitus, smoking, and any current medication. Patients underwent 24-h ambulatory blood pressure monitoring and blood sampling. Lipid peroxidation was measured in the plasma and erythrocytes as 8-isoprostane or malondialdehyde (MDA), respectively. Antioxidant capacity was measured as ferric reducing ability of plasma (FRAP) in the plasma and as reduced/oxidized glutathione (GSH/GSSG ratio) in erythrocytes. (Na + K)-ATPase activity and fatty acids were determined in erythrocyte membranes. Hypertensives had higher levels of plasma 8-isoprostane, erythrocyte MDA, and relative percentage of saturated membrane fatty acids, but lower plasma FRAP levels, erythrocyte GSH/GSSG ratio, (Na + K)-ATPase activity and relative percentage of unsaturated membrane fatty acids, compared with normotensives. Day-time systolic and diastolic blood pressures correlated positively with lipid peroxidation parameters, but negatively with (Na + K) ATPase activity. These findings suggest that the modulation of (Na + K)-ATPase activity may be associated with changes in the fatty acid composition induced by oxidative stress and provide evidence of a role for this enzyme in the pathophysiology of essential hypertension. PMID- 17410407 TI - Purification and kinetic properties of glutathione reductase from bovine liver. AB - Glutathione reductase (GR, NADPH: oxidized glutathione oxidoreductase, EC 1.6.4.2) catalyzes the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH) using NADPH as reducing cofactor. The aim of the present work was to purify and characterize GR from bovine liver. GR was purified using 2', 5' ADP-Sepharose 4B and DEAE-Sepharose Fast Flow columns. The enzyme has been purified 5456-fold and with a yield of 38.4%. The molecular and catalytic properties of bovine liver GR have been studied. Optimum temperature and pH was found to be 50 degrees C and 7, respectively. The activation energy of the reaction catalyzed by the enzyme was 9.065 kcal/mole. The molecular weight of the enzyme was found to be 55 kDa by SDS-PAGE. Kinetic characterization of bovine liver GR was also investigated, Km(NADPH) 0.063 +/- 0.008 mM and Km(GSSG) 0.154 +/- 0.015 mM were determined. It is accepted that parallel lines observed in these double reciprocal plots obeys Ping Pong mechanism and we have showed this in our steady state study. According to our results of statistical analysis, the Ping Pong mechanism is a suitable model since the loss function is less than the other mechanisms. However, competitive inhibition by a product could be accepted in sequential mechanisms but not in a Ping Pong mechanism. In this study, kinetic data are consistent with a branching reaction mechanism previously proposed for GR from other sources by other studies. PMID- 17410408 TI - Propolis protects CYP 2E1 enzymatic activity and oxidative stress induced by carbon tetrachloride. AB - Induction of CYP 2E1 by carbon tetrachloride (CCl(4)) is one of the central pathways by which CCl(4) generates oxidative stress in hepatocytes. Experimental liver injury was induced in rats by CCl(4) to determine toxicological actions on CYP 2E1 by microsomal drug metabolizing enzymes. In this report, ethanolic extract of propolis at a dose of 200 mg/kg (po) was used after 24 h of toxicant administration to validate its protective potential. Intraperitoneal injection of CCl(4) (1.5 ml/kg) induced hepatotoxicity after 24 h of its administration that was associated with elevated malonyldialdehyde (index of lipid peroxidation), lactate dehydrogenase and gamma-glutamyl transpeptidase release (index of a cytotoxic effect). Hepatic microsomal drug metabolizing enzymes of CYP 2E1 showed sharp depletion as assessed by estimating aniline hydroxylase and amidopyrine N demethylase activity after CCl(4) exposure. Toxic effect of CCl(4) was evident on CYP 2E1 activity by increased hexobarbitone induced sleep time and bromosulphalein retention. Propolis extract showed significant improvement in the activity of both enzymes and suppressed toxicant induced increase in sleep time and bromosulphalein retention. Choleretic activity of liver did not show any sign of toxicity after propolis treatment at a dose of 200 mg/kg (id). Histopathological evaluation of the liver revealed that propolis reduced the incidence of liver lesions including hepatocyte swelling and lymphocytic infiltrations induced by CCl(4). Electron microscopic observations also showed improvement in ultrastructure of liver and substantiated recovery in biochemical parameters. Protective activity of propolis at 200 mg/kg dose was statistically compared with positive control silymarin (50 mg/kg, po), a known hepatoprotective drug seems to be better in preventing hepatic CYP 2E1 activity deviated by CCl(4). These results lead us to speculate that propolis may play hepatoprotective role via improved CYP 2E1 activity and reduced oxidative stress in living system. PMID- 17410410 TI - Sickness in the system: the health costs of the harvest. AB - Cherie Moraga's play, Heroes and Saints, and Helena Maria Viramontes' novel, Under the Feet of Jesus, offer readers perspectives on the lives of migrant farm workers in California that challenge the moral imagination and conscience. Both focus on health hazards of pesticides and on the often prohibitive difficulty of getting health care for those who fall ill as a result of exposure. This paper offers a reflection on the direct political and moral appeal these works present to readers who may not see or acknowledge the sacrifices sustained by those whose undercompensated labors are an integral part of our food systems. PMID- 17410409 TI - Low-temperature deposition of rutile film on biomaterials substrates and its ability to induce apatite deposition in vitro. AB - Low-temperature deposition of crystalline titania films on intrinsically bioinert materials to induce the bioactivity is of practical interest, not only because it meets the demand of providing organic biomaterials with bioactivity, which cannot tolerate high-temperature thermal treatments, but also because it reserves abundant Ti-OH groups facilitating the apatite deposition. In this paper, rutile films with thickness varied from 0.1 microm to 1.7 microm were deposited on commercially available pure titanium substrates from 1.5 M titanium tetrachloride aqueous solution kept at 60 degrees C for 3-60 h. The rutile films grew to give a preferred (101) crystalline plane in the X-ray diffraction pattern. After soaking in a simulated body fluid of the Kokubo solution (SBF) for 2 days, the rutile films with thickness over 0.6 microm were covered with a layer of apatite. All the films with various thickness induced apatite deposition in SBF after soaking for 5 days. The bioinert polytetrafluoroethylene (PTFE) was also found to exhibit remarkable in vitro bioactivity as to induce apatite deposition from SBF within 2 days, after depositing the rutile film on the surface. PMID- 17410411 TI - Regularities in eyewitness identification. AB - What do eyewitness identification experiments typically show? We address this question through a meta-analysis of 94 comparisons between target-present and target-absent lineups. The analyses showed that: (a) correct identifications and correct-nonidentifications were uncorrelated, (b) suspect identifications were more diagnostic with respect to the suspect's guilt or innocence than any other response, (c) nonidentifications were diagnostic of the suspect's innocence, (d) the diagnosticity of foil identifications depended on lineup composition, and (e) don't know responses were nondiagnostic with respect to guilt or innocence. Results of diagnosticity analyses for simultaneous and sequential lineups varied for full-sample versus direct-comparison analyses. Diagnosticity patterns also varied as a function of lineup composition. Theoretical, forensic, and legal implications are discussed. PMID- 17410412 TI - Decreased ghrelin-induced GH release in thyrotoxicosis: comparison with GH releasing peptide-6 (GHRP-6) and GHRH. AB - In thyrotoxicosis GH response to several stimuli is impaired, but there is no data on ghrelin-induced GH release in these patients. Ghrelin is a potent GH secretagogue and it also increases glucose levels in men. The aim of this study was to evaluate the effects of ghrelin (1 microg/kg), GHRP-6 (1 mug/kg) and GHRH (100 microg), i.v., on GH levels in 10 hyperthyroid patients and in 8 controls. Glucose levels were also measured during ghrelin and GHRP-6 administration. In control subjects and hyperthyroid patients peak GH (microg/l; mean +/- SE) values after ghrelin injection (controls: 66.7 +/- 13.6; hyper: 19.3 +/- 2.4) were significantly higher than those obtained after GHRP-6 (controls: 26.7 +/- 5.1; hyper: 12.6 +/- 1.3) and GHRH (controls: 13.5 +/- 4.3; hyper: 5.3 +/- 1.3). There was a significant decrease in GH responsiveness to ghrelin, GHRP-6 and GHRH in the hyperthyroid group compared to controls. In control subjects and hyperthyroid patients basal glucose (mmol/l) values were 4.5 +/- 0.1 and 4.7 +/- 0.2, respectively. There was a significant increase in glucose levels 30 min after ghrelin injection (controls: 4.9 +/- 0.1; hyper: 5.2 +/- 0.2), which remained elevated up to 120 min. When the two groups were compared no differences in glucose values were observed. GHRP-6 administration was not able to increase glucose levels in both groups. Our data shows that GH release after ghrelin, GHRP 6 and GHRH administration is decreased in thyrotoxicosis. This suggests that thyroid hormone excess interferes with GH-releasing pathways activated by these peptides. Our results also suggest that ghrelin's ability to increase glucose levels is not altered in thyrotoxicosis. PMID- 17410413 TI - Immunohistochemical properties of silent corticotroph adenoma and Cushing's disease. AB - Proopiomelanocortin processing in corticotroph cells is known to be operated by prohormone convertase (PC) 1/3 which is activating several pro-proteins and prohormones by intracellular limited proteolysis processing. In this study, we hypothesized that PC1/3 expression differs between Cushing's disease (CD) and silent corticotroph adenoma (SCA), and investigated whether PC1/3 expression is involved in the adrenocorticotropin (ACTH) silence of SCA. We performed immunohistochemical analysis of pituitary adenoma specimens for six adenohypophysial hormones, PC1/3 and chromogranin A (CgA). Subjects for this study consisted of 12 anterior pituitary adenomas of CD (1 male, 11 female; 14-70 years old) and 31 non-functioning adenomas (23 male, 8 female; 32-71 years old).ACTH immunoreactivity was observed in all of CD and three of 31 non functioning adenomas. The three cases diagnosed as SCA were also positive for growth hormone and follicle-stimulating hormone. Cushing's adenomas and SCAs were all positive for PC1/3. PC1/3-positive cells did not always colocalize with ACTH but some of them colocalized with CgA in SCAs. Even if PC1/3 is not present in corticotroph cells, PC1/3 immunoreactivity in SCA may originate from CgA-positive cells. We conclude that immunohistochemistry for PC1/3 is not helpful for differential diagnosis between CD and SCA in clinical practice, though the regulation of PC1/3 expression is likely to be an important etiological factor in ACTH silence of SCA. The diversity of immunohistochemical properties of SCA leads us to speculate that it is not a single entity and may be a general diagnostic term for adenomas of varying etiology. PMID- 17410414 TI - Beyond false beliefs: the development and psychometric evaluation of the perceptions of children's theory of mind measure-experimental version (PCToMM-E). AB - The Perceptions of Children's Theory of Mind Measure (Experimental version; PCToMM-E) is an informant measure designed to tap children's theory of mind competence. Study one evaluated the measure when completed by primary caregivers of children with autism spectrum disorder. Scores demonstrated high test-retest reliability and correlated with verbal mental age and ToM task battery performance. No ceiling effects were observed. In addition, caregivers accurately predicted their children's ToM task battery performance. In study two the scores of primary caregivers of typically developing children demonstrated high test retest reliability and distinguished children on the basis of age and developmental status. Ceiling effects were not evident until late childhood. The utility of the PCToMM-E and directions for future research are discussed. PMID- 17410415 TI - Prevalence of pervasive developmental disorder in Down's syndrome. AB - The frequencies of pervasive developmental disorder (PDD) in Down's syndrome (DS) have been reported from 1% to 11%. However, it is not clear if the frequency of this co-occurrence is higher or lower than in other mental retardations. We study a large sample of DS population, finding a PDD frequency of 15.6%, with 5.58% of autism (eight males and two females) and 10.05% of PDD non autism (nine males and nine females. The meaning of this frequency is discussed. PMID- 17410416 TI - Brief report: data on the Stanford-Binet Intelligence Scales (5th ed.) in children with autism spectrum disorder. AB - The Fifth Edition of the Stanford-Binet Intelligence Scales (SB5; Roid, G. H. (2003). Stanford Binet intelligence scales (5th ed.). Itasca, IL: Riverside Publishing) is relatively new, with minimal published research on general populations and none with special populations. The present study provides information on the cognitive profiles of children with ASD (N=63) and on the whether the abbreviated battery is representative of the full scale. A high percentage of the children had significantly stronger nonverbal (vs. verbal) skills. This pattern was not related to Full Scale IQ, age or diagnostic subgroup. IQs derived from the abbreviated battery accounted for a large proportion of the variance in FSIQ relative to comparable abbreviated batteries. However, caution is warranted when using the abbreviated battery, as it misrepresents actual ability in a small percentage of cases. PMID- 17410417 TI - Anger management style and endogenous opioid function: is gender a moderator? AB - This study explored possible gender moderation of previously reported associations between elevated trait anger-out and reduced endogenous opioid analgesia. One hundred forty-five healthy participants underwent acute electrocutaneous pain stimulation after placebo and oral opioid blockade in separate sessions. Blockade effects were derived reflecting changes in pain responses induced by opioid blockade. Hierarchical regressions revealed that elevated anger-out was associated with smaller pain threshold blockade effects (less opioid analgesia) in females, with opposite findings in males (interaction p < .001). Similar marginally significant interactions were noted for blockade effects derived for nociceptive flexion reflex threshold, pain tolerance, and pain ratings (p < .10). Anger-in was also associated negatively with pain threshold blockade effects in females but not males (interaction p < .05). Across genders, elevated anger-in was related to smaller pain tolerance blockade effects (p < .01). Overlap with negative affect did not account for these opioid effects. The anger-in/opioid association was partially due to overlap with anger-out, but the converse was not true. These findings provide additional evidence of an association between trait anger-out and endogenous opioid analgesia, but further suggest that gender may moderate these effects. In contrast to past work, anger in was related to reduced opioid analgesia, although overlap with anger-out may contribute to this finding. PMID- 17410418 TI - Depression history, stress, and pain in rheumatoid arthritis patients. AB - This study examined the role of past episodes of depression on pain reports for patients with rheumatoid arthritis (RA) before and during stress induction. A history of major depressive episodes was assessed by diagnostic interviews for 138 RA patients, 74 who later participated in a set of laboratory procedures designed to induce interpersonal stress. Patients were evaluated by a rheumatologist and then asked to report joint and bodily pain throughout the laboratory study. We found that RA patients with a history of two or more episodes of major depression had more pain at baseline, and exhibited higher pain in response to the stress induction than did RA patients with either only one episode or no history of depression. Such findings provide new insight in the dynamic relationships between depression, stress, and pain. PMID- 17410420 TI - Implementation of evidence-based practice in child welfare: service provider perspectives. AB - Implementation of evidence-based practices (EBP) in child welfare is a complex process that is often fraught with unanticipated events, conflicts, and resolutions. To some extent, the nature of the process, problems, and solutions may be dependent on the perspectives and experiences of a given stakeholder group. In order to better understand the implementation process in the child welfare system, we interviewed comprehensive home-based services (CHBS) case managers who were actively engaged in implementing an EBP to reduce child neglect in a state youth services system. Six primary factors were identified as critical determinants of EBP implementation: (1) Acceptability of the EBP to the caseworker and to the family, (2) Suitability of the EBP to the needs of the family, (3) Caseworker motivations for using the EBP, (4) Experiences with being trained in the EBP, (5) Extent of organizational support for EBP implementation, and (6) Impact of EBP on process and outcome of services. These factors reflect two broader themes of attitudes toward or assessments of the EBP itself and experiences with learning and delivering the EBP. Eventual implementation is viewed as the consequence of perseverance, experience, and flexibility. PMID- 17410419 TI - Prospective study of attitudinal and relationship predictors of sexual risk in the multicenter AIDS cohort study. AB - We examined the influence of attitudes concerning HIV transmission, safe sex, and sexual sensation seeking, as well as negotiated risk reduction with primary partners, on the proportion of unprotected sexual partners (%UASP) among men who have sex with men (MSM). Participants were 263 HIV-seropositive and 238 HIV seronegative MSM in the Multicenter AIDS Cohort Study between 1999 and 2003 who completed a 20-item attitude survey twice. Behavioral data were collected concurrently and 6-12 months after each survey. Among seropositives, decreased HIV concern and increased safer sex fatigue were associated with higher %UASP at 6 and 12 months. Among seronegatives, increased %UASP at 12 months was associated with safer sex fatigue. At 6 months and 12 months, risk reduction agreements were associated with increased %UASP among seronegatives in seroconcordant monogamous relationships, reflecting their abandonment of condoms in such partnerships. We conclude that HIV prevention efforts should target modifiable attitudes (reduced concern about HIV and safer sex fatigue) and increases in sexual risk-taking of MSM, particularly among HIV+ men having sex with serodiscordant partners. PMID- 17410421 TI - Quantitative PCR technique for the identification of microrearrangements of the AZFc region. AB - PURPOSE: The AZFc region spans about 3.5 Mb and contains many amplicons causing recombination events. Several papers have reported the occurrence of AZFc partial deletions resulting from non allelic homologous recombination (NAHR) ("gr-gr", "b1-b3" or "b2-b3" deletions), particularly in infertile patients. DAZ genes are present in 4 copies and rearrangements involve a modification of the number of DAZ genes. METHODS: In addition to STS plus/minus PCR, we developed a quantitative technique using real time PCR (Q-PCR) to determine the number of DAZ genes. Fourteen DNA controls were selected to validate the use of Q-PCR to detect AZFc microrearrangements, and sperm DNA samples from 30 fertile men were studied. RESULTS: Rearrangements of 14 controls were well identified with Q-PCR, and 2 AZFc partial deletions were detected in fertile men (1 "gr-gr" and 1 "b2-b3"). CONCLUSION: Q-PCR represents a well-adapted method to detect microrearrangements of the Y-chromosome, complementary to STS analysis. PMID- 17410422 TI - Novel mutations found in two genes of thai patients with isolated methylmalonic acidemia. AB - Molecular genetic analysis of three patients diagnosed with isolated methylmalonic acidemia (MMA) revealed that one was mut (0) MMA, with a mutation in the MUT gene encoding the L: -methylmalonyl-CoA mutase (MCM), and two were cblB MMA, with mutations in the MMAB gene required for synthesizing the deoxyadenosylcobalamin cofactor of MCM. The mut (0) patient was homozygous for a novel nonsense mutation in MUT, p.R31X (c.167C --> T), and heterozygous for three previously described polymorphisms, p.K212K (c.712A --> G), p.H532R (c.1671A --> G), and p.V671I (c.2087G --> A). The new MMAB mutation, p.E152X (c.454G --> T), was found to be homozygous in one cblB patient and heterozygous in the other patient, who also had four intron polymorphisms in this gene. PMID- 17410423 TI - Getting the social organism thinking: strategy for systems change. AB - The ability of community researchers/practitioners to facilitate systems change is constrained by social power--particularly the capacity to shape ideology [S. Lukes (1974). Power: A radical view. Hampshire: MacMillan] and frequently power molds ideologies which undermine systems thinking. Following what Mills [C. W. Mills, (1959). The sociological imagination. New York: Oxford University Press] (termed the "sociological imagination", this article makes the case for a strategy of systems change that promotes an integrated focus on systems and their constituent individuals. Both of these components are understood to continuously shape each other. The social imagination is introduced as a way to conceptualize the intersection between individuals' conceptions of systems and the ways that systems work to form individual identities and perceptions of social reality. Examples of attempts at systems change from community organizing and public health are used to illustrate both common fallacies and potential future directions for systems change efforts. PMID- 17410424 TI - Systems change as an outcome and a process in the work of community collaboratives for health. AB - The widespread development of comprehensive community initiatives that aim to improve community health is driven by the need to change the systems charged with delivering the services and creating the policies related to a variety of health outcomes. Georgia's Family Connection initiative is the nation's largest statewide network of community collaboratives for health, with collaboratives operating in 159 counties. Data on community context, collaborative processes, engagement in systems change, and changes in programs and activities implemented, gathered consistently at the collaborative level over 3 years, will be used to answer the following questions. How do community contexts and the structure and processes of collaboratives affect implementation of systems change? How do systems changes affect intermediate outcomes such as the type of programs offered in a community? Longitudinal change in systems change and program implementation is described and significant predictors of between-collaborative variation in longitudinal change for each outcome are identified. PMID- 17410425 TI - Changing systems by changing individuals: the incubation approach to systems change. AB - This article describes and evaluates the implementation of an innovative approach to systems change, the incubation approach, which was developed on a systems change project designed to increase the capacity of multiple systems (e.g., law enforcement, child protection, domestic violence, mental health, early education) to respond to children's exposure to violence. The incubation approach encourages change agents to collaborate with project staff to gently nurture, or "incubate," feasible and warranted change in target systems. Project staff gain concrete commitment from motivated and accessible change agents and collaborate with those agents to implement change actions. This approach works well with committed, executive-level change agents in target systems, with stable systems that have low turnover and well-integrated subsystems, and when seed funds are provided to key organizations. PMID- 17410426 TI - A comparison of repetitive behaviors in Aspergers Disorder and high functioning autism. AB - In this study we compared 33 IQ and age matched pairs of individuals with Aspergers Disorder (ASP) and high functioning autism (HFA) on measures of repetitive behavior. On the Repetitive Behavior Scale-Revised (RBS-R), the ASP and HFA groups showed no differences in RBS-R Intensity score (severity) score or Frequency score (number of problems present). This suggests that the two groups are similar with respect to the intensity or severity of repetitive behaviors and the presence of repetitive behaviors. At the item level there were no differences on scales typically associated with autism (Stereotyped Behavior) and ASP (Restricted Interests). Similarly, there were no differences between the groups on the Aberrant Behavior Checklist Stereotypy scale. These findings add to the body of literature showing that HFA and ASP fail to differ with respect to repetitive behaviors. The implications of the findings for neurobiologic and genetic studies are discussed. PMID- 17410427 TI - Heavy metal concentrations in diet and livers of Black-crowned Night Heron Nycticorax nycticorax and Grey Heron Ardea cinerea chicks from Pyeongtaek, Korea. AB - This study presents concentrations of iron, manganese, zinc, copper, lead and cadmium in diet and livers of Black-crowned Night Heron Nycticorax nycticorax and Grey Heron Ardea cinerea chicks from Pyeongtaek, Gyeonggi-do, Korea. Heavy metal concentrations of heron chicks were not related to concentrations in the diet. Copper concentrations were significantly greater in the diet of Black-crowned Night Herons (geometric mean = 13.6 wet microg/g) than Grey Herons (7.45 wet microg/g), other metal concentrations did not differ between the diet of two species. Manganese (respectively 3.20 wet microg/g, 1.41 wet microg/g) and cadmium (respectively 13.4 wet microg/kg, 1.41 wet microg/kg) concentrations were higher in livers of Black-crowned Night Heron chicks than Grey Heron chicks, but zinc, iron, copper and lead concentrations in livers did not differ in between two herons. The essential elements were at background levels, however copper concentrations were relatively higher than previously reported from Korea. Lead and cadmium concentrations were within background levels for herons. PMID- 17410428 TI - Multivariate logistic regression for familial aggregation in age at disease onset. AB - Familial aggregation studies seek to identify diseases that cluster in families. These studies are often carried out as a first step in the search for hereditary factors affecting the risk of disease. It is necessary to account for age at disease onset to avoid potential misclassification of family members who are disease-free at the time of study participation or who die before developing disease. This is especially true for late-onset diseases, such as prostate cancer or Alzheimer's disease. We propose a discrete time model that accounts for the age at disease onset and allows the familial association to vary with age and to be modified by covariates, such as pedigree relationship. The parameters of the model have interpretations as conditional log-odds and log-odds ratios, which can be viewed as discrete time conditional cross hazard ratios. These interpretations are appealing for cancer risk assessment. Properties of this model are explored in simulation studies, and the method is applied to a large family study of cancer conducted by the National Cancer Institute-sponsored Cancer Genetics Network (CGN). PMID- 17410429 TI - Giant adenoma arising in the lower esophagus with adenocarcinoma. PMID- 17410430 TI - A splice-site mutation in exon 4 of the APC gene in a family with attenuated familial adenomatous polyposis. PMID- 17410431 TI - Papillary endothelial hyperplasia of the liver: a case report and review of the literature. PMID- 17410432 TI - Multiple myeloma presenting as gastric polyposis. PMID- 17410433 TI - Association between Helicobacter pylori infection in mothers and birth weight. AB - Helicobacter pylori infection may cause intrauterine growth restriction (IUGR). However, it is unknown whether the growth of children from H. pylori-infected mothers is also affected or whether transmission of infection from mother to child occurs. This study aimed to determine if maternal H. pylori infection was associated with IUGR and low birth weight in a mouse model, and whether transmission of infection from mother to infant occurs. Female C57BL/6 mice were inoculated with H. pylori (n = 18) or water (control; n = 18) via gavage. Mice were mated at 6 weeks postinfection, with half of the mice sacrificed after 2 weeks of gestation. The remaining mice gave birth and a third of the litter was weighed and sacrificed at birth, during milk feeding (1.5 weeks), and during solid feeding (4 weeks). Stomachs of all mice and whole foetuses were cultured for the presence of H. pylori. There were no differences in litter size or foetus weight between control and H. pylori-infected mice. Pups from infected mothers had a lower weight during milk feeding (control, 5.91 +/- 0.23 g; H. pylori, 4.59 +/- 0.16 g; p < 0.05) and solid feeding (control, 12.73 +/- 0.58 g; H. pylori, 10.01 +/- 1.02 g; p < 0.05). H. pylori was not detected by culture in the pups at any age. H. pylori infection in mothers was associated with a decrease in infant weight during milk feeding and after weaning. Transmission of infection from mother to infant was not detected by culture, suggesting that decreased baby weight may be due to decreased milk supply or altered nutrition from the mother. PMID- 17410434 TI - Impairment of gastric ulcer healing by alendronate, a nitrogen-containing bisphosphonate, in rats. AB - Bisphosphonates such as alendronate have been developed as antiresorptive agents capable of treating diseases related to bone remodeling. In the present study, we examined the effect of alendronate on the healing of acetic acid-induced gastric ulcers in rats and investigated the mechanism involved in this action both in vivo and in vitro using the rat gastric epithelial cell line (RGM1). Acetic acid induced gastric ulcers healed spontaneously, with up-regulation of COX 2/prostaglandin E2 production as well as expression of vascular endothelium derived growth factor (VEGF) and basic fibroblast growth factor (bFGF) in ulcerated mucosa. The healing of ulcers was impaired by indomethacin (2 mg/kg, s.c.) or alendronate (60 mg/kg, p.o.) given once daily for 7 days, starting 3 days after acid application. Indomethacin, but not alendronate, inhibited mucosal prostaglandin E2 production. Alendronate as well as indomethacin decreased the protein expression of both VEGF and bFGF in ulcerated mucosa, resulting in a reduction of angiogenesis in the ulcer base. Supplementation of recombinant bFGF significantly reverted the delay in ulcer healing caused by alendronate. On the other hand, the size of cell-free areas in RGM1 cells in vitro decreased with time after wound induction, and this process was promoted by epidermal growth factor (EGF; 10 ng/ml). Co-incubation with alendronate (1 mM) did not affect the spontaneous healing but significantly suppressed the accelerated wound healing caused by EGF. These results suggest that alendronate impairs the healing of gastric ulcers in rats, and this effect may be related to down-regulation of VEGF and bFGF, the important growth factors for vascularization/granulation, as well as suppression of the stimulatory action of EGF on epithelial proliferation/migration. PMID- 17410435 TI - Prevention of complications and symptomatic recurrences in diverticular disease with mesalazine: a 12-month follow-up. AB - In uncomplicated diverticular disease, treatment is aimed at relieving symptoms. The aim of the present study was to evaluate the efficacy of mesalazine for symptomatic relief of uncomplicated diverticular disease of the colon. Two hundred sixty-eight consecutive eligible outpatients (122 male, 146 female; age, 66.1 years; range, 31-81 years) were enrolled in four treatment schedules in a randomized fashion: Group R1 (66 patients), rifaximin, 200 mg bid; Group R2 (69 patients), rifaximin, 400 mg bid; Group M1 (67 patients), mesalazine, 400 mg bid; and Group M2 (66 patients), mesalazine, 800 mg bid. Treatments were administered for 10 days every month for 12 months. Clinical evaluations were performed at admission and at 3-month intervals for 12 months considering 12 clinical variables (upper and lower abdominal pain/discomfort, tenesmus, diarrhea, abdominal tenderness, fever, bloating, general illness, nausea, emesis, dysuria, bleeding) graded as 0 = no symptoms, 1 = mild, 2 = moderate, and 3 = severe. The Global Symptomatic Score (GSS) was calculated using the sum of each symptom score. Two hundred forty-four patients completed the 12- month study; 24 were discontinued (14 treated with rifaximin and 10 treated with mesalazine) either as voluntary dropouts or because they developed side effects and/or complications. Group M2 demonstrated a lower frequency of many symptoms after 6 and 12 months of treatment; the mean GSS was significantly lower in Group M2 after 6 and 12 months of therapy by both intention-to-treat and per-protocol analyses. Patients treated with mesalazine (Groups M1+M2) had a lower GSS than subjects treated with rifaximin (Groups R1+R2) during the 12-month follow-up period. We conclude that cyclic administration of mesalazine is effective for symptomatic relief of uncomplicated diverticular disease of the colon. Some symptoms showed greater improvement with mesalazine, 800 mg bid, than with the other treatment schedules. PMID- 17410436 TI - The role of zinc and metallothionein in the dextran sulfate sodium-induced colitis mouse model. AB - Zinc (Zn) and its binding protein metallothionein (MT) have been proposed to suppress the disease activity in ulcerative colitis. To determine the role of Zn and MT in the dextran sulfate sodium (DSS)-induced model of colitis in mice, a DSS dose-response study was conducted in male C57BL/6 wild-type (MT+/+) and MT null (MT-/-) mice by supplementing 2%, 3%, and 4% DSS in the drinking water for 6 days. In the intervention study, colitis was induced with 2% DSS, Zn (24 mg/ml as ZnO) was gavaged (0.1 ml) daily, concurrent with DSS administration, and the disease activity index (DAI) was scored daily. Histology, MT levels, and myeloperoxidase (MPO) activity were determined. DAI was increased (P<0.05) by 16% and 21% with 3% and 4% concentrations of DSS, respectively, compared to 2%, evident after 5 days of DSS administration. MPO activity was increased in MT+/+ compared to MT-/- mice and those receiving DSS. Zn administration had a 50% (P<0.05) lower DAI compared to DSS alone. Zn partially prevented the distal colon of MT+/+ by 47% from DSS-induced damage compared to MT-/- mice. MT did not prevent DSS-induced colitis and Zn was partially effective in amelioration of DSS induced colitis. PMID- 17410437 TI - Stem cell factor/c-kit receptor signaling enhances the proliferation and invasion of colorectal cancer cells through the PI3K/Akt pathway. AB - In this study, we examined the role of c-kit receptor (KIT) signal transduction on the proliferation and invasion of colorectal cancer cells. We found that c-kit was expressed in 2 colorectal cancer cell lines as determined by RT-PCR, Western blot, and flow cytometry. In KIT-positive lines, KIT was activated by stem cell factor (SCF). SCF enhanced cellular proliferation of positive lines as demonstrated by the WST-1 proliferation assay. Furthermore, SCF enhanced the invasive ability of KIT-positive cell lines. SCF stimulation upregulated p44/42 mitogen-activated protein kinase (MAPK) and Akt as shown by Western blot. We examined the roles played by p44/42 MAPK and phosphatidylinositol 3-kinase (PI3K)/Akt pathways in proliferation and invasion. PI3K/Akt activity strongly correlated with proliferation and invasion and p44/42 MAPK was correlated with only invasion. In conclusion, the SCF-enhanced proliferation and invasion of KIT positive colorectal cancer cells is achieved mainly through the PI3K/Akt pathway. PMID- 17410438 TI - Hypermethylation of SFRP2 as a potential marker for stool-based detection of colorectal cancer and precancerous lesions. AB - DNA methylation is a key mechanism of colorectal carcinogenesis. Analysis of aberrantly methylation in stool DNA might provide a novel strategy for noninvasive detection of colorectal cancer (CRC). To explore the feasibility of this approach, we have assessed the methylation status of secreted frizzled related protein gene 2 (SFRP2) in stool samples from patients with CRC with respect to a series of healthy individuals and patients with benign colorectal diseases, using methylation-specific polymerase chain reaction. Methylated SFRP2 occurs in 94.2%, 52.4%, 37.5%, and 16.7% of patients with CRC, adenomas, hyperplstic polyps, and ulcerative colitis, respectively. Of the 24 normal individuals, only 1 revealed methylated DNA. The pilot study revealed that aberrant methylated SFRP2 could be detected frequently in stools from patients with CRC and precancerous lesions. Methylation testing of fecal DNA may be a simple, promising, and noninvasive screening tool for colorectal neoplasia. PMID- 17410439 TI - Clinical utility of double-balloon enteroscopy for small intestinal bleeding. AB - Until the development of wireless capsule endoscopy (CE) and double-balloon enteroscopy (DBE), it was extremely difficult to examine the entire small intestine. To assess the usefulness of DBE for diagnosing suspected small intestinal bleeding, we retrospectively compared the diagnoses and treatments of cases before and after its introduction at one hospital. Between September 2003 and December 2005, 21 consecutive patients with suspected small intestinal bleeding underwent DBE at Tokai University Hospital (group A), and subsequently 2 were excluded from the study after being diagnosed with bleeding from a diverticulum and an angiodysplasia in the ascending colon, respectively. For comparison, inpatients who were negative for gastrointestinal bleeding on colonoscopy and gastroscopy between May 1998 and August 2003 were reviewed and 27 consecutive patients who had not undergone DBE were selected as the control group (group B). All patients had been diagnosed negative for a source of bleeding on more than one colonoscopy and gastroscopy. There were no significant differences between the two groups in terms of age, gender, history of blood transfusion, blood hemoglobin value on admission, or symptoms. The diagnostic yield of DBE in identifying the source of bleeding was 78.9%: six cases of small intestinal ulcers, five cases of angiodysplasia, two cases of hard submucosal tumor (SMT), one case of small pulsating SMT, and one case of small intestinal cancer. DBE was also used to successfully treat three cases of angiodysplasia with argon plasma coagulation. In the control group, conventional investigations, including enteroclysis, angiography, Meckel scan, scintigraphy with technetium-labeled red blood cells, and/or push enteroscopy, were performed in 88.9%, 29.6%, 29.6%, 55.6%, and 25.9%, respectively. The overall diagnostic yield of the conventional approaches was only 11.1% (P < 0.01), comprising a Meckel's diverticulum, a polyp, and an angiodysplasia. We conclude that DBE can be used to diagnose suspected small intestinal bleeding and to treat some cases, such as angiodysplasia. PMID- 17410440 TI - Efficacy of T2 in active Crohn's disease: a prospective study report. AB - To date few therapies have been shown to reliably prevent the evolution of Crohn's disease (CD). The traditional Chinese medicine, Tripterygium wilfordii Hook F (TWHF), has both immunomodulatory and anti-inflammatory activities. Our aim was to investigate the potential efficacy of T2, the major constituent of extracts of TWHF, in inducing remission of active CD. Twenty adult patients with active CD were enrolled to be treated with T2 pills (60 mg daily) for 12 weeks. Plasma levels of C-reactive protein (CRP), tumor necrosis factor (TNF)-alpha, and interleukin (IL)-1beta were measured at entry and every 2 weeks thereafter until week 12. At each visit the CD Activity Index (CDAI) was calculated. The CD Endoscopic Index of Severity was measured at entry and week 12. Sixteen patients completed the study. A significant decrease in serum levels of CRP, TNF-alpha, and IL-1beta occurred rapidly after commencement of treatment. CDAI scores showed a rapid decline during the first 8 weeks and reached their lowest at week 10. Endoscopic improvements were observed at week 12. In conclusion, T2 appears to be effective for the treatment of mildly or moderately active CD. Further controlled studies are warranted for this promising drug. PMID- 17410441 TI - Telmisartan, an angiotensin II type 1 receptor blocker, controls progress of nonalcoholic steatohepatitis in rats. AB - The term nonalcoholic steatohepatitis (NASH) has recently been proposed to identify a fatty liver disease accompanied by diffuse fatty infiltration and inflammation. However, no drug therapy has been established for NASH as yet. In the present study, we demonstrate the effect of the angiotensin II type 1 receptor antagonist telmisartan on the development of NASH in a rat model. Telmisartan, but not the angiotensin receptor antagonist valsartan, markedly attenuated hepatic steatosis, inflammation, and fibrosis in these rats. The quantitative parameters of steatosis, inflammation, and fibrosis were also ameliorated by treatment with telmisartan. Compared with telmisartan, the peroxisome proliferator-activated receptor-gamma agonist pioglitazone attenuated hepatic steatosis and fibrosis of the liver to a similar degree. However, telmisartan, but not pioglitazone, dramatically decreased both subcutaneous and visceral fat. In conclusion, these results indicated that telmisartan should be the drug of first choice for the treatment of patients with NASH. PMID- 17410442 TI - CARD15 status and familial predisposition for Crohn's disease and colonic gene expression. AB - Familial disposition and mutations in the Caspase Recruitment Domain 15 (CARD15) have been associated with an increased risk for Crohn's disease (CD). This study investigated whether these risk factors correlate with colonic gene expression profiles generated by DNA-microarray technology. Tissue specimens from descending colon were obtained during colonoscopy from 45 CD patients (18 from areas with inflammation and 27 from noninflamed areas). Gene profiling analysis was performed using the Human Genome U133 Plus 2.0 GeneChip Array. Patients were classified according to their CARD15 status. Hybridization data were analyzed with dChip software. Nine patients with either one or two CARD15 mutations had no differentially expressed genes, compared to 36 patients with wild- type CARD15. There was only one differentially expressed EST between 8 patients who had familial disposition for inflammatory bowel disease (IBD) and 36 who did not, but hierarchical cluster analysis did not show group homogeneity. We conclude that gene expression profiling of mucosal biopsies from the descending colon of patients with CD could not be correlated with CARD15 status or with familial disposition for IBD. PMID- 17410443 TI - Platelet-activating factor inactivator (rPAF-AH) enhances liver's recovery after paracetamol intoxication. AB - Platelet-activating factor (PAF) is a potent endogenous phospholipid modulator of diverse biological activities, including inflammation. The aim of this study was to investigate the effects of PAF inactivator, recombinant PAF acetylhydrolase (rPAF-AH) on post-paracetamol treatment functional outcome of the liver in the rat. Fifty male Wistar rats were divided into two groups: the control group received by gastric tube a toxic dose of paracetamol (3.5 g/kg body weight) and the rPAF-AH-treated group received the same dose of paracetamol followed by a dose of rPAF-AH (10 mg/kg body weight) intraperitoneally. The animals were sacrificed at 8, 16, 24, 32, and 40 hr after paracetamol treatment. APAP was found to cause acute hepatic injury, evident by alterations of biochemical (serum enzymes: ALT, AST, and ALP) and liver histopathological (degree of inflammation and apoptosis) indexes, which was followed by liver regeneration evident by three independent indexes ([(3)H]thymidine incorporation into hepatic DNA, liver thymidine kinase activity, and hepatocyte mitotic index). Hepatic levels of malondialdehyde (MDA) and serum cholesterol/HDL cholesterol fraction were also measured as parameters of oxidant-antioxidant balance. The positive effects of rPAF-AH were expressed by (1) a reduction of oxidative stress, (2) a large decrease in hepatic injury, and (3) a reduction of regenerating activity. These results suggest that PAF plays an important role in paracetamol-induced liver injury and regeneration. Furthermore, PAF inactivator enhances liver's recovery and attenuates the severity of experimental liver injury, providing important means of improving liver function following paracetamol intoxication. PMID- 17410444 TI - Ascites and kidney transplantation: case report and critical appraisal of the literature. AB - Ascites is an ominous finding after kidney transplantation. It occurs either as a result of technical complication of the transplant procedure or from medical reasons that include portal hypertension, graft rejection and other causes (1, 2). Here in are described a case of ascites that occurred after successful kidney transplantation. Kidney re-transplantation alone or combined heart and kidney transplantation were possible options for him. Finally, a review of published cases of ascites that occurred after kidney transplantation and treatment of putative nephrogenic ascites (NA) is presented. PMID- 17410445 TI - Hepatitis C hypervariable region 1: association of reduced selection pressure in african americans with treatment failure. AB - In a prospective therapeutic trial, features of the hepatitis C quasispecies were investigated as possible markers of therapeutic response. Individuals chronically infected with hepatitis C genotype 1 received antiviral therapy consisting of alpha-interferon plus ribavirin. The study targeted the most rapidly evolving segment of the viral genome, hypervariable region 1 within the envelope-2 gene. Among individuals failing to clear virus in response to therapy, significant differences were observed between quasispecies of African-American and Caucasian subjects. While distance measures for synonymous substitutions were similar between racial subgroups, measures of distance at the amino acid level (nonsynonymous substitutions) varied significantly. Taken together, the observed patterns of variability corresponded to reduced host selection pressure against hypervariable region 1 in African-American nonresponders. Reduced selection pressure was present at baseline and persisted through treatment and follow-up, suggesting population stratification of host factors that influence selection pressure on hepatitis C virus. PMID- 17410446 TI - The influence of hemorrhagic shock on rat liver regeneration after partial hepatectomy: serum aminotranspherases, mitochondrial function, and hepatocellular replication studies. AB - This study was designed to evaluate the influence of hemorrhagic shock on hepatic regeneration in rats submitted to partial hepatectomy. The experimental protocol included 26 male Wistar rats, randomly assigned to 4 groups: GI: simulated operation; GII: 30% hepatectomy without hemorrhagic shock; GIII: only hemorrhagic shock; GIV: 30% hepatectomy associated with hemorrhagic shock. The methodologies used were: determination of aminotranspherases plasma levels; analysis of mitochondrial respiration, membrane potential and osmotic swelling; and markers of hepatocellular replication. Aminotranspherases increased only in GIV. There were no differences in mitochondrial respiration. Mitochondrial membrane potential decreased only in the GIV. There were no differences in mitochondrial swelling among the groups; cellular replication markers increased significantly in the Groups II and IV but without difference between these two groups. Despite the conditions imposed on the organism by hemorrhagic shock, the hepatic regenerative capacity is preserved in animals submitted to partial hepatectomy. PMID- 17410447 TI - Correlation between findings on MRCP and gadolinium-enhanced MR of the liver and a survival model for primary sclerosing cholangitis. AB - Primary sclerosing cholangitis is a chronic, progressive disease of inflammation and fibrosis of the bile ducts. The ability to predict survival is important for appropriate management and treatment decisions. The purpose of this study was to examine the relationship between specific findings on the enhanced magnetic resonance imaging (MRI) examination of the liver and the corresponding magnetic resonance cholangiopancreatogram (MRCP) and a survival model for primary sclerosing cholangitis (PSC), the Mayo Risk Score. During a five-year period, 47 patients with primary sclerosing cholangitis were identified who had a complete MRI/MRCP examination. The extent of anatomical changes of the biliary tree and the degree of peribiliary enhancement in the hepatic parenchyma were compared with the Mayo Risk Score for each patient. Peribiliary enhancement was present to a varying extent in 38 of 47 cases of PSC. Peribiliary enhancement 3 min after gadolinium administration had a weak correlation with the Mayo Risk Score (analysis of variance P<0.01, Pearson correlation r=0.37). No statistically significant relationship between the severity of extrahepatic or intrahepatic duct changes and the Mayo Risk Score was found (analysis of variance P=0.24, P=0.38, respectively). Although biliary tree changes on MRCP aid in the diagnosis of PSC, they do not correlate with survival, as predicted by the Mayo Risk Score. Peribiliary enhancement on MRI of the liver is a finding occurring to a variable extent in primary sclerosing cholangitis but does not correlate significantly with survival. PMID- 17410448 TI - Clinical course of hepatitis B virus infection in renal allograft recipients. AB - The outcome of renal transplantation is adversely affected by hepatitis B virus infection. We retrospectively analyzed data of 1,251 renal transplant recipients, 20 of whom were hepatitis B surface antigen positive and hepatitis B virus DNA negative at the time of renal transplantation. Hepatitis B virus reactivation was seen in 14 of the 20 patients at a mean time of 16.3+/-7.1 months after transplantation. All patients with hepatitis B virus reactivation after transplantation were treated with lamivudine, biochemical, and serologic response was achieved in 13 of 14 patients at a mean time of 7.0+/-1.1 months. Seven of 13 patients experienced a breakthrough at a mean time of 9.2+/-6.2 months. Three of the 20 patients died at a mean time of 57.0+/-38.5 months after transplantation. Our data demonstrated that chronic immunosuppression is associated with a significantly high risk of hepatitis B virus reactivation in renal transplant recipients and hepatitis B reactivation does not increase the likelihood of graft rejection or patient mortality after renal transplantation. PMID- 17410449 TI - Effect of withdrawal of nonsteroidal anti-inflammatory drug use on ileal pouch disorders. AB - NSAID use has been shown to exacerbate disease activity of inflammatory bowel disease. The detrimental effect of NSAIDs on the ileal pouch has not been characterized. To study the effect of withdrawal of NSAID use on ileal pouch disorders. The study consisted of a cohort of 17 symptomatic patients seen in the Pouchitis Clinic who had ulcerative colitis and ileal pouch-anal anastomosis with chronic (>6 months) daily use of NSAIDs. The patients were treated by withdrawing NSAID use. The Pouchitis Disease Activity Index (PDAI) consisting of symptom, endoscopy and histology scores, and Cleveland Global Quality of Life, Irritable Bowel Disease Quality of Life, and Short Inflammatory Bowel Disease Questionnaire scores were measured before and after a 4-week intervention. The cohort consisted of 11 patients with chronic refractory pouchitis (65%), 2 with acute pouchitis (12%), 1 with cuffitis (6%), 1 with cuffitis and chronic refractory pouchitis (6%), and 2 with irritable pouch syndrome (12%). The withdrawal of NSAID use alone resulted in a significant reduction in the mean PDAI scores of -3.6 +/- 3.0 (p<0.02) and a significant improvement in mean quality-of-life scores (p<0.05). Patients with pouch disorders who regularly used NSAIDs appeared to benefit from the complete cessation of such agents, suggesting an association between NSAID use and pouch disorders. PMID- 17410450 TI - Determinants of Abnormal Supine Reflux in 24-Hour pH Recordings. AB - A previous study showed that supine gastroesophageal reflux occurs mainly in the early nighttime period and is greater if the patient retires within 2 hr of a meal. We analyzed 306 consecutive esophageal pH studies between January and July 2003. Ninety-two patients had abnormal supine reflux. The recumbent period was divided into quarters (Q1-Q4) and reflux parameters were calculated for each period. Reflux was maximal in Q1, and decreased progressively from Q1 to Q4 (P<0.001). Median time between evening meal and retiring was no different from that for 44 upright gastroesophageal reflux patients. Patients retiring within 2 hr of a meal had a higher percentage supine reflux time (P=0.012), but no increase in reflux was observed in those retiring within 3 hr. Supine reflux occurs maximally in the early nighttime period. Although not fully explained as a postprandial phenomenon, these patients should avoid going to bed within 3 hr of a meal. PMID- 17410451 TI - Helicobacter pylori infection and severity of reflux-induced esophageal disease in a cohort of patients with columnar-lined esophagus. AB - The relationship between Helicobacter pylori infection and reflux-induced esophageal diseased is controversial. We examined esophageal disease severity in patients with columnar-lined esophagus and compared results between patients with and without Helicobacter pylori infection. Medical records of 1000 patients diagnosed with columnar-lined esophagus were examined. Endoscopic and histological findings of reflux-induced esophageal disease were compared between H. pylori-positive and H. pylori-negative patients. Four hundred twenty-nine patients (42.9%) showed evidence of H. pylori status, of whom 239 (55.7%) were positive and 190 (44.3%) negative. There were no significant differences in length of columnar-lined segment (P = 0.305), frequency of associated esophagitis (P = 0.583), or presence of gastroduodenal inflammation (P = 0.335, P = 0.131) between the two groups. Histological grade of esophageal disease severity was similar between them, with no statistically significant differences (P = 0.231). We conclude that in patients with established columnar-lined esophagus, there appears to be no difference in severity of reflux-induced esophageal disease between those with and those without H. pylori infection. PMID- 17410452 TI - Effect of fermented soybean meal on intestinal morphology and digestive enzyme activities in weaned piglets. AB - The objective of the current experiment was to investigate the effect of Bacillus subtilis fermented soybean meal (FSBM) on intestinal morphology and digestive enzyme activities in piglets. Sixty crossbred (Duroc x Landrance x Yorkshire) piglets at an average weight of 8.62 kg were randomly allotted to two treatments and fed diets containing soybean meal (SBM) and FSBM, respectively. At the end of the experimental period, six piglets of each treatment were slaughtered humanly to collect the contents and tissue samples at three different locations in small intestine. Light microscopy and scanning electron indicated that piglets fed FSBM had a higher (P<0.05) villus height at three different locations of small intestine and had la ower (P<0.05) crypt depth in the duodenum compared to piglets fed SBM. The results showed that activities of total protease and trypsin at the duodenum and jejunum of piglets fed with FSBM increased (P<0.05) compared with the control. The trypsin activity in the pancreas of piglets decreased (P<0.05) when they were fed with FSBM. The results showed that FSBM improved intestinal morphology and digestive enzyme activities in weaned piglets. PMID- 17410453 TI - Arterial embolization for hemorrhage caused by hepatic arterial injury. AB - Fewer than 10% of patients with major liver trauma have life-threatening bleeding. Laparoscopic operations, endoscopic procedures, and percutaneous interventions such as drainages, vascular or tumor sclerosis, biopsies, and transjugular intrahepatic portocaval shunt (TIPS) have increased the number of iatrogenic vascular and bile duct injuries. Due to its therapeutic success, arterial embolization (AE) has become the standard treatment for late complications of hepatic injury. In some cases, this procedure may be used as the first approach on accidental or iatrogenic arterial trauma and in several hepatic arterial diseases. The result of this method depends on physician experience, size of the wound, and stability of the patient. Persistent hemorrhage and rebleeding may be treated with a new AE or an operation. Precise indication and a correctly performed AE are key factors for a successful treatment. Rebleeding episodes are a rare occurrence, which may be treated with AE as well. PMID- 17410454 TI - The effect of a silybin-vitamin e-phospholipid complex on nonalcoholic fatty liver disease: a pilot study. AB - Oxidative stress leads to chronic liver damage. Silybin has been conjugated with vitamin E and phospholipids to improve its antioxidant activity. Eighty-five patients were divided into 2 groups: those affected by nonalcoholic fatty liver disease (group A) and those with HCV-related chronic hepatitis associated with nonalcoholic fatty liver disease (group B), nonresponders to treatment. The treatment consisted of silybin/vitamin E/phospholipids. After treatment, group A showed a significant reduction in ultrasonographic scores for liver steatosis. Liver enzyme levels, hyperinsulinemia, and indexes of liver fibrosis showed an improvement in treated individuals. A significant correlation among indexes of fibrosis, body mass index, insulinemia, plasma levels of transforming growth factor-beta, tumor necrosis factor-alpha, degree of steatosis, and gamma-glutamyl transpeptidase was observed. Our data suggest that silybin conjugated with vitamin E and phospholipids could be used as a complementary approach to the treatment of patients with chronic liver damage. PMID- 17410455 TI - Postinfantile giant cell hepatitis due to hepatitis E virus along with the presence of autoantibodies. PMID- 17410457 TI - Can soluble triggering receptor expressed on myeloid cells (sTREM-1) be considered an anti-inflammatory mediator in the pathogenesis of peptic ulcer disease? AB - Soluble triggering receptor expressed on myeloid cells (sTREM-1) is a novel mediator involved in the pathogenesis of peptic ulcer disease. To investigate the potential role of sTREM-1 in the anti-inflammatory response in chronic gastritis, sTREM-1 was compared with other anti-inflammatory mediators of gastritis. Forty patients with dyspepsia were enrolled: 20 with peptic ulcer and 20 controls without any macroscopic abnormalities. All patients were examined by endoscopy; gastric juice was aspirated and biopsy specimens were collected from the antrum and corpus of the stomach. sTREM-1, interleukin (IL)-8, and IL-10 were estimated by enzyme immunoassays. Median sTREM-1 in patient controls and in patients with peptic ulcer disease was 3.91 and 44.27 pg/ml, respectively (P=0.006). Respective values of IL-8 were 1856.97 and 2030.66 pg/ml (P=0.023); those of IL-10 were 16.92 and 18.43 pg/ml (NS). The odds ratio for the presence of peptic ulcer in the event of a concentration of sTREM-1 higher than 15 pg/ml was 23.22 (95% CI, 2.58-208.62; P=0.002). A positive correlation was found between the ratios of IL 8/sTREM-1 and IL-8/IL-10 (r (s), + 0.365; P=0.021). In conclusion, sTREM-1 is an independent factor for the generation of peptic ulcer disease and might behave as an anti-inflammatory mediator in chronic gastritis. PMID- 17410456 TI - Leukocytapheresis (LCAP) in the management of chronic active ulcerative colitis- results of a randomized pilot trial. AB - Recent studies suggest that leukocytapheresis with Cellsorba is a valuable therapy for ulcerative colitis after failure of conventional treatment. In this study the potential of leukocytapheresis to induce remission in refractory chronic colitis under the conditions of European treatment guidelines was investigated. The therapeutic benefit of leukocytapheresis in the maintenance of remission was additionally elucidated. Twenty patients were treated weekly for 5 weeks. A significant decrease in the activity index was observed. Fourteen patients achieved clinical remission, and mucosal healing was observed endoscopically in six patients. After randomization these 14 patients in remission entered a second period of either monthly leukocytapheresis or no further treatment. In both groups steroids were tapered down. After 6 months, only one patient in the control group remained in remission, in contrast to five of eight patients in the leukocytapheresis group. In conclusion, leukocytapheresis may offer a therapeutic option in the induction and the maintenance of remission in chronic active ulcerative colitis. PMID- 17410458 TI - Hepatic subcapsular hematoma secondary to intrahepatic pseudoaneurysm following cholecystectomy. PMID- 17410459 TI - HFE gene mutation, chronic liver disease, and iron overload In Turkey. AB - We aimed to determine the relationships between iron overload and HFE gene mutation in chronic liver disease in Turkey. One hundred thirteen chronic liver disease patients and 138 healthy controls were evaluated regarding their clinical, biochemical, and genetic parameters. Each group was divided into two subgroups according to transferrin saturation (TS) (45% and >45%). HFE gene mutation was analyzed by the PCR-RFLP method. C282Y homozygote, heterozygote, and wild-type mutation rates were 1.7%, 0%, and 98.3% in patients and 0%, 1.4%, and 98.6% in controls, respectively. H63D homozygote, heterozygote, and wild-type mutation rates were 1.8%, 24.7%, and 73.5% in patients and 1.4%, 24%, and 74.6% in controls, respectively. Mutation rates were not statistically different in patients with high and normal TS. Iron overload was positively correlated with biochemical activity and Child-Pugh score (P < 0.05). In multivariate analysis, H63D homozygotic mutation was an independent factor for the development of hepatocellular carcinoma (P = 0.004). We conclude that C282Y mutation is very rare in Turkey. Iron overload is not related to H63D mutation but is positively correlated with biochemical activity and Child-Pugh score in chronic liver diseases. PMID- 17410460 TI - Analysis of the T1288R mutation of the Wilson disease ATP7B gene in four generations of a family: possible genotype-phenotype correlation with hepatic onset. AB - Wilson disease, an autosomal recessive disorder due to mutations of the ATP7B gene, is characterized by copper accumulation and toxicity in the liver and subsequently in other organs, mainly the brain and cornea. A new missense mutation (T1288R) of the ATP7B gene has recently been discovered in a Wilson disease patient in our laboratory. The aim of the present study was to analyze clinical and genetic features of more generations of the family of the patient in which the new mutation T1288R was discovered. A total of 19 subjects were studied; in particular, four generations of the patient's family were analyzed. The ATP7B gene was analyzed by single-strand conformational polymorphism followed by direct sequencing. Two brothers presented a clinical diagnosis of Wilson disease with an hepatic phenotype and a genotype characterized by the homozygotic mutation T1288R. The heterozygotic mutation T1288R was found in seven subjects belonging to all four generations. The present study represents the first screening for a Wilson disease mutation through four generations of a nonconsanguineous family. All the patients with the homozygotic T1288R mutation in the present pedigree presented an hepatic phenotype without a neurological presentation. Consequently, a genotype-phenotype correlation could be hypothesized, although further studies are necessary to clarify this topic. PMID- 17410461 TI - Acid-suppressive effects of various regimens of omeprazole in Helicobacter pylori negative CYP2C19 homozygous extensive metabolizers: which regimen has the strongest effect? AB - To achieve more potent and long-lasting acid suppression, omeprazole was administered for 7 days in 5 regimens: 10, 20, and 40 mg once daily (od), and 10 and 20 mg twice daily (bid), in 7 healthy Helicobacter pylori-negative CYP2C19 homozygous extensive metabolizers, and intragastric pH was continuously measured. The median intragastric pH and percent time pH > 4.0 for 24 hours increased dose dependently with 10, 20, and 40 mg od. Ten and 20 mg bid wre comparable to 20 and 40 mg od, respectively. Concerning percent time pH > 4.0 in the nighttime (20:00 8:00 hours), 20 mg bid was significantly superior to 40 mg od (P < .05). In 4 of the 5 regimens, all 7 subjects had nocturnal acid breakthrough, whereas with 20 mg bid it occurred in only 3. We concluded that, considering nighttime acid suppression, omeprazole 20 mg bid had the strongest effect. PMID- 17410462 TI - The beneficial effect of N-acetylcysteine and ciprofloxacin therapy on the outcome of ischemic fulminant hepatic failure. PMID- 17410464 TI - Small early invasive colon cancer near the splenic flexure successfully diagnosed by intraoperative magnifying endoscopy. PMID- 17410463 TI - Bowel habit reference values and abnormalities in young Iranian healthy adults. AB - The purpose of this study was to estimate the prevalence of self-reported, ROME II-defined constipation and determine the average defecation frequency and stool types in the Iranian population. A self-reported questionnaire was distributed to 1045 participants, including items intended to identify the presence of ROME II defined functional constipation and the dominant form of stool based on the Bristol Scale. The weekly mean bowel movement frequency in men and women was 12.5 +/- 7.3 and 13.8 +/- 8.0, respectively (p < 0.05). A total of 87.4% of participants had a stool frequency of between 3 and 21 times per week. The prevalence of functional constipation was 32.9%, whereas only 9.6% of participants reported themselves to be constipated (level of agreement kappa = 0.21, 95% confidence interval: 0.15 to 0.27). Soft or formed stool was reported in 75.7% of individuals. Functional constipation is common in the Iranian population, but its diagnosis could not rely on subjective patient complaints. Despite a higher average of bowel frequency, the previously reported normal range of defecation frequency can be applied for the Iranian population. PMID- 17410465 TI - Sarcoidosis of gastrointestinal tract: a rare disease. AB - Gastrointestinal (GI) tract sarcoidosis is an uncommon form of sarcoidosis. The GI tract can be involved as an isolated disease as a part of systemic sarcoidosis. Clinical manifestations of esophageal, gastric, small bowel, colon, and appendicular sarcoidosis are discussed in this review. The differential diagnosis of GI sarcoidosis is extensive. Other granulomatous diseases of the GI tract, like tuberculosis, fungal infections, parasitic diseases, inflammatory bowel disease, and Whipple's disease, should be excluded before making the diagnosis of GI sarcoidosis. Corticosteroid therapy is effective in treatment of GI sarcoidosis. Surgical intervention may be necessary in patients with bowel obstruction, perforation, or massive hemorrhage. PMID- 17410466 TI - Serum concentrations of insulin-like growth factor-I (igf-I) as a marker of liver fibrosis in patients with chronic hepatitis C. AB - Liver biopsy was until recently the only way of evaluating liver fibrosis. Noninvasive tests for hepatic fibrosis, without potential risks, are desired by clinicians as well as patients. Insulin-like growth factor-I (IGF-I) synthesis is disturbed in liver fibrosis and reflects the severity of the clinical stage. We assessed serum IGF-I levels in patients with chronic hepatitis C (CHC) to correlate with liver fibrosis and antiviral therapy. Forty patients with CHC and persistently abnormal alanine aminotransferase values were enrolled and treated with peginterferon alpha-2a 180 microg per week plus ribavirin for 24 (n=20) or 48 (n=20) weeks. All patients underwent liver biopsy before treatment (METAVIR fibrosis stage F0, n=13; F1-F2, n=14; F3, n=7; F4, n=6). Serum IGF-I was measured at baseline, at the end of treatment period, and 24 weeks after finishing treatment. Mean IGF-I values were significantly lower in patients with advanced fibrosis (F4, 65.9+/-17.9 ng/mL) than in the others (F0, 145.2+/-47.1; F1-F2, 150.3+/-89.6; and F3, 121.4+/-35.2 ng/mL; P < .05). Serum IGF-I levels increased during combined therapy, being this increment markedly higher in patients with sustained virologic response. In conclusion, IGF-I synthesis is disturbed in CHC and reflects the severity of the liver fibrosis. Combined therapy improves serum IGF-I levels. IGF-I could represent a good, noninvasive marker of liver fibrosis. PMID- 17410467 TI - Preventive effects of rebamipide on NSAID-induced gastric mucosal injury and reduction of gastric mucosal blood flow in healthy volunteers. AB - The precise mechanisms of acute damage and the role of gastric mucosal blood flow in gastric mucosal injury induced by nonsteroidal anti-inflammatory drugs (NSAIDs) remain uncertain. The aim of this study was to evaluate the preventive effect of rebamipide on gastric mucosal injury and reduction of gastric mucosal blood flow (GMBF) after ibuprofen administration. Twenty healthy volunteers were randomized two groups. The rebamipide group took ibuprofen, 1800 mg/day, and rebamipide, 100 mg t.i.d., for 7 days. The placebo group took ibuprofen, 1800 mg/day. The numbers of gastric ulcer subjects were three in the placebo group and zero in the rebamipide group. The mean modified Lanza score after ibuprofen administration was significantly higher in the placebo group than the rebamipide group (2.9+/-1.7 vs. 1.3+/-1.0, respectively; P=0.032). The GMBF of the placebo group was significantly decreased at antrum from baseline, from 2.8+/-0.5 to 2.0+/-0.5 tissue perfusion units (P=0.005). There was no difference in GMBF change in the rebamipide group. Gastric mucosal injury was correlated with GMBF reduction in antrum (r=-0.677, P=0.001). In conclusion, it is suggested that the decrease in GMBF may have been associated with NSAID-induced gastric mucosal injury, and rebamipide may have prevented NSIAD-induced gastric mucosal injury by maintaining GMBF in healthy subjects. PMID- 17410468 TI - Gastrointestinal bleeding secondary to hepatic artery involvement of microscopic polyangiitis: case report and review of the literature. PMID- 17410469 TI - Meta-analysis of neuropsychological symptoms of adolescents and adults with PKU. AB - Phenylketonuria (PKU; OMIM 261600) is an autosomal recessive inborn error of phenylanaline metabolism. PKU is characterized by deficient or defective phenylalanine hydroxylase activity and persistantly increased levels of the essential amino acid phenylalanine in the circulation. The present article examines current understanding of the etiology of PKU, along with a meta-analysis examining neuropsychological and intellectual presentations in continuously treated adolescents and adults. Patients with PKU differed significantly from controls on Full-Scale IQ, processing speed, attention, inhibition, and motor control. Future research utilizing an integrative approach and detailed analysis of specific cognitive domains will assist both the scientist and clinician, and ultimately the patient. PMID- 17410470 TI - Defining normalcy of the somatotropic axis: an attainable goal? AB - The diagnoses of acromegaly and dwarfism require biochemical confirmation of abnormal GH and IGF-1 concentrations. The same parameters are used for therapeutic decisions, i.e. initiation or termination of particular treatments. Therefore, reliable and epidemiologically and statistically proven criteria of normalcy for GH and IGF-1 are required for these tasks to be accomplished. Despite major progress in all these areas, the definition of what constitutes "normalcy" of the somatotropic axis is still lacking. Using an example of acromegaly, we discuss the contradictions and the uncertainties of the biochemical diagnosis of this disease. PMID- 17410471 TI - IGF-I measurements in the monitoring of GH therapy. AB - Growth hormone replacement therapy has been used regularly in adult Growth hormone deficiency since the availability of recombinant GH in the 1980's. GH replacement improves quality of life, bone turnover markers, cardiovascular risk markers and adverse body composition. Originally, GH doses in replacement regimes were determined by weight and surface area and dose increases based on body composition outcomes analogous to pediatric practice. These regimens led to significant side effects related to excess GH, arthralgias, headaches and peripheral edema and IGF-I levels above the upper limit of the reference range. Newer treatment regimes therefore account for known factors affecting serum GH and IGF-I levels, i.e. age, gender, estrogen replacement and pre-treatment IGF-I levels. Monitoring is now via clinical symptomatology combined with serum total IGF-I levels, potentially this avoids excessive GH exposure and allows monitoring of compliance and dose titration. There is a lack of data relating IGF-I to biological endpoints, but analysis suggests that dose titration of IGF-I to the upper half of the age and gender related reference range is acceptable. The use of reliable IGF-I assays and extensive age and gender related reference ranges is necessary and centralized monitoring is preferable. Free IGF-I and bioavailable IGF-I measurements are available but their use in the monitoring of GH replacement remains to be determined. PMID- 17410472 TI - The relative effect of household and workplace smoking restriction on health status among Chinese Americans living in New York City. AB - Households and workplaces are the predominant location for exposure to secondhand smoke. The purpose of this study is to examine the association between health status and smoking restrictions at home and work and to compare the relative effect of household and workplace smoking restrictions on health status. This study uses data from a cross sectional representative probability sample of 2,537 Chinese American adults aged 18-74 living in New York City. The analysis was limited to 1,472 respondents who work indoors for wages. Forty-three percent of respondents reported a total smoking ban at home and the workplace, 20% at work only, 22% home only, and 15% reported no smoking restriction at home or work. Nonsmokers [corrected] who live under a total household smoking ban only or both a total household and total workplace ban were respectively 1.90 and 2.61 times more likely to report better health status compared with those who reported no smoking ban at work or home. Before the NYC Clean Indoor Air Act second-hand smoke (SHS) exposure among this immigrant Chinese population at home and work was high. This study finds that household smoking restrictions are more strongly associated with better health status than workplace smoking restrictions. However, better health status was most strongly associated with both a ban at work and home. Public health efforts should include a focus on promoting total household smoking bans to reduce the well-documented health risks of SHS exposure. PMID- 17410473 TI - Pattern of settlement and natural chimerism in the colonial urochordate Botryllus schlosseri. AB - Colonies of the cosmopolitan urochordate Botryllus schlosseri that share one or both alleles at a single allorecognition locus (Fu/HC) and come into tissue contacts, may fuse and form a mixed entity, a chimera. Botryllus populations worldwide exhibit unprecedented extensive polymorphism at this locus, a result that restricts fusions to kin encounters. This study aims to compare spatiotemporal configurations in source and introduced B. schlosseri populations, residing on natural and man-made substrata, respectively. By using four microsatellite loci, we tested genetic consanguinity of colonies settled naturally along spatial vectors on both, natural (native populations) and man made (introduced) substrates. Four populations were studied. Results revealed that B. schlosseri colonies, on both substrate types, assemble in groups of relatives that share similar microsatellite profiles. We suggest that this pattern of settlement promotes the formation of chimeras, which evoke conflicting interactions: cooperation between different somatic cell lines that constitute the colonial soma and competition between germ cells that inhabit the chimera gonads. Under natural conditions, the chimera may allow genetic flexibility that depends on joint genomic fitness of its partners. This is probably one of the life history characteristics that led to the worldwide distribution success of this species. PMID- 17410474 TI - Delimiting the role of ethical reasoning in epidemiology. PMID- 17410475 TI - Polymorphism distribution of prion protein codon 117, 129 and 171 in Taiwan. AB - Prion diseases compass transmissible spongiform neurodegenerative diseases from various causes, including the genetic and infectious ones. We investigated the prevalence of codon 117, 129 and 171 polymorphism in prion protein (PrP) in Taiwanese, mainly for the sake of the informative absence of this genetic distribution. Our subjects were 419 aged ones of Han ethic origin. We evaluated the PrP gene (PRNP) polymorphism by restriction fragment length polymorphism, after amplification of their genomic DNAs by polymerase chain reactions with specific primers, digested by restriction enzyme PvuII (for codon 117), NspI (for codon 129), and BbvI (for codon 171), respectively, and confirmed by nucleotide sequencing. All of the subjects were homozygotes at codon 117 (Ala/Ala, gca/gca) and 171 (Asn/Asn, aac/aac). There were no valine homozygotes (Val/Val) in our 419 subjects, and nine subjects (2.1%) showed methionine-valine heterozygosity (Mal/Val, atg/gtg). The methionine homozygotes (Met/Met) comprised the major population (97.9%), and the prevalence of distribution is different to that seen in Caucasians. The almost 100% conservation of the domain from codon 117 to 171 implies the warranty of PrP in cellular functions. The high prevalence of Met/Met alleles in Taiwan did not imply an increased risk of CJD, and the genetic susceptibility of CJD by codon 129 of PrP may be still elusive for the infectivity. PMID- 17410476 TI - Corrected incidences of co-morbidities - a statistical approach for risk assessment in anesthesia using an AIMS. AB - OBJECTIVE: In anesthesia and intensive care logistic regression analysis are often used to generate predictive models for risk assessment. Strictly seen only independent variables should be represented in such prognostic models. Using anesthesia-information-management-systems a lot of (depending) information is stored in a database during the preoperative ward round. The objective of this study was to evaluate a statistical algorithm to process the different dependent variables without losing the information of each variable on patient's conditions. METHOD: Based on data about prognostic models in anesthesia an iterative statistical algorithm was initiated to summarize dependent variables to subscores. Seven subscores out of several preoperative variables were calculated corresponding to the proper incidence and the correlation to the occurrence of intraoperative cardiovascular events was evaluated. After that first step logistic regression was used to build a predictive model out of the seven subscores, 10 patient-related, and two surgery-related variables. Performance of the prognostic model was assessed using analysis of discrimination and calibration. RESULT: Four out of seven subscores together with age, type and urgency of surgery are represented in the prognostic model to predict the occurrence of intraoperative cardiovascular events. The prognostic model demonstrated good discriminative power with an area under the ROC curve (AUC) of 0.734. CONCLUSION: Due to reduced calibration, the clinical use of the prediction model is limited. PMID- 17410477 TI - Serum calcium and breast cancer risk: results from a prospective cohort study of 7,847 women. AB - Experimental and epidemiological studies suggest that calcium-regulating hormones -parathyroid hormone (PTH) and vitamin D--may be associated with breast cancer risk. No prospective cohort study has investigated the association between pre diagnostic calcium levels and subsequent risk of breast cancer. We have examined this in a cohort of 7,847 women where serum calcium levels and established risk factors for breast cancer had been assessed at baseline. During a mean follow-up of 17.8 years, 437 incident breast cancer cases were diagnosed. Incidence of breast cancer was calculated in different quartiles of serum calcium levels and a Cox's proportional hazards analysis was used to obtain corresponding relative risks (RR), with a 95% confidence interval (CI), adjusted for potential confounders. In premenopausal women, serum calcium levels were inversely associated with breast cancer risk in a dose-response manner. The adjusted RR (95% CI) of breast cancer was in the 2nd calcium quartile 0.91 (0.65-1.30), in the 3rd quartile 0.89 (0.60-1.31), and in the 4th quartile 0.56 (0.32-0.98), as compared to the 1st calcium quartile. In postmenopausal overweight women (BMI > 25), breast cancer risk was higher in calcium quartiles 2-4 as compared to the 1st quartile. Our findings may have implications for primary prevention of breast cancer and for the management of asymptomatic primary hyperparathyroidism. PMID- 17410478 TI - Statin therapy depresses total body fat oxidation in the absence of genetic limitations to fat oxidation. AB - Cholesterol lowering drugs are associated with myopathic side effects in 7% of those on therapy, which is reversible in most, but not all patients. This study tested the hypothesis that total body fat oxidation (TBFO) is reduced by statins in patients with genetic deficiencies in FO, determined by white blood cells (FOwbc) and by molecular analysis of common deficiencies, and would cause intolerance in some patients. Six patients on statin therapy without myopathic side effects (tolerant) and 7 patients who had previously developed statin induced myopathic symptoms (intolerant) (age = 58 +/- 8.25 yrs, ht. = 169 +/- 11 cm, and wt. = 75.4 +/- 14.2 kg) were tested for TBFO (Respiratory Exchange Ratio, RER) pre- and during exercise. FOwbc was not significantly different between tolerant and intolerant (0.261 +/- 0.078 vs. 0.296 +/- 0.042 nmol/h per 10(9) wbc), or normals (0.27 +/- 0.09 nmol/h per 10(9) wbc) and no common molecular abnormalities were found. Pre-exercise RER (0.73 +/- 0.05 vs. 0.84 +/- 0.05) was significantly lower in the intolerant group and the VO2 at RER = 1.0 (1.27 +/- 0.32 vs. 1.87 +/- 0.60 L/min) greater than the tolerant. Post-exercise lactates were not different between groups. Although dietary fat intake was not different, blood lipoprotein levels, particularly triglycerides were 35% lower in tolerant than previously intolerant. TBFO and blood lipoproteins were reduced in tolerant patients in spite of the absence of genetic limitations, but not in the intolerant group as hypothesized. Although not conclusive, these data suggest the need for a prospective study of the effects of statins on fat oxidation. PMID- 17410479 TI - Left ventricular function in professional football players evaluated by tissue Doppler imaging and strain imaging. AB - BACKGROUND: Long-term regular exercise is associated with physiologic and morphologic cardiac alterations. Tissue Doppler Imaging(TDI) and Strain Myocardial Imaging(SI) are new tools in the evaluation systolic and diastolic myocardial function. We sought to compare TDI and SI findings in professional football players and age adjusted sedentary controls to assess the effect of regular athletic training on myocardial function. METHODS: Transthoracic echocardiography, M-mode, 2-D measurements, Doppler derived mitral-tricuspid annular velocities, reconstructed spectral pulsed wave tissue Doppler velocities, strain and strain rate imaging of seven different myocardial regions were obtained from 24 professional football players and age, sex and weight adjusted 20 controls. RESULTS: Age, body surface area, blood pressure and heart rate were comparable between 2 groups. Football players had significantly increased LV mass, mass index (due to both higher wall thickness and end-diastolic diameter), end-systolic and end-diastolic volume, left atrial diameter and decreased transmitral diastolic late velocity. In athletes TDI analysis showed significantly increased mitral annulus septal TDI peak early diastolic(e) velocity(0.22 +/- 0.04 vs. 0.19 +/- 0.04 m/s, P < 0.05), lateral TDI peak e velocity (0.19 +/- 0.03 vs. 0.16 +/- 0.02 m/s, P < 0.05) and lateral TDI e/a ratio (1.96 +/- 0.41 and 1.66 +/- 0.23, P < 0.05). In SI analysis mid septal walls (1.71 +/- 0.23 in athletes and 1.49 +/- 0.25 in controls, P < 0.05) and mid lateral walls (1.55 +/- 0.28 and 1.34 +/- 0.25 respectively, P < 0.05) peak systolic strain rate values differences were found to be increased in athletes. CONCLUSIONS: Professional football playing is associated with morphologic alteration in left ventricle and left atrium and improvement in left ventricle diastolic function which can be detected by TDI. Strain rate imaging may be a new tool to define subtle change in systolic left ventricular function in "athletes heart" which cannot be determined in standard echocardiographic parameters. PMID- 17410480 TI - Update on the contribution of galactomannan for the diagnosis of invasive aspergillosis. AB - The diagnosis of invasive fungal infections (IFI) remains a challenge, particularly for diseases caused by filamentous fungi such as Aspergillus species. Unfortunately, many patients affected by these conditions are not identified before autopsy. Therefore, there is a need for new diagnostic methods for IFI. Galactomannan is a soluble antigen released during hyphal growth in tissues. A commercially available sandwich ELISA assay that detects galactomannan has been used in Europe for many years and is now approved for use in the USA. The test has an excellent negative predictive value in the detection of invasive aspergillosis (IA) in high-risk patients. In addition, it is more sensitive than culture and allows IA to be diagnosed before clinical manifestations occur. However, false-negative and false-positive results in certain populations are the main limitations to its use. The purpose of this review is to summarize the current knowledge about galactomannan testing in patients at risk for IA. PMID- 17410481 TI - [CT-imaging findings in idiopathic hypereosinophilic syndrome]. PMID- 17410492 TI - [Medicine and art--a bridge between two worlds]. PMID- 17410498 TI - [Meeting of the Austrian Image Processing Group]. PMID- 17410499 TI - Initial experience with the medtronic ADVANTAGE valve prosthesis in the mitral position--clinical outcome and analysis of complications. AB - OBJECTIVE: The Medtronic ADVANTAGE prosthetic heart valve is a bileaflet mechanical heart valve which has two main design modifications. The prosthesis has an enlarged central orifice to improve blood flow characteristics and an asymmetric butterfly pivot recess with expanded outflow component designed to enhance the blood flow washing through the pivot region. This report summarizes the initial experience with 55 patients who underwent isolated mitral valve replacement. METHODS: We prospectively followed 55 patients undergoing mitral valve replacement with the Medtronic ADVANTAGE prosthesis in a single center study. All patients were operated on via a median sternotomy and a left atrial approach. RESULTS: The age of the patients at implant ranged from 42 years to 77 years with a mean of 61.8 years. Forty-five percent of the patients were male. Seventy-five percent of the patients were in NYHA functional class III or IV prior to valve replacement. The diameters of the implanted prostheses were as follows: 25 mm in 2 patients, 27 mm in 12 patients, 29 mm in 20 patients, and 31 mm in 21 patients. Coronary artery bypass grafting was performed with valve replacement in 16.4 % of patients and additional tricuspid repair in 10.9 %. After one year, 97.6 % and after two years 100 % of the included patients were in NYHA functional class I or II. All patients had Coumadin (warfarin) therapy postoperatively with a target INR range of 2.5 to 3.5, or 3.0 to 4.0. There were 4 thromboembolic events (7.3 %) in the early period and 2 late events (2.6 %). Episodes of valve thrombosis were not seen during the follow-up, which consisted of 76.5 patient years. The hemodynamic performance was favorable and within clinically acceptable ranges. The incidence of valve-related mortality and morbidity in this preliminary study was extremely low, indicating good clinical results comparable to those reported for other bileaflet valves. CONCLUSION: The ADVANTAGE valve is a safe and effective option for mitral valve replacement with a very low incidence of valve-related complications. PMID- 17410500 TI - Mechanical aortic and mitral valve replacement in infants and children. AB - BACKGROUND: The aim of this study was to evaluate early and late outcomes after mechanical systemic heart valve replacement in pediatric patients. METHODS: Between October 1981 and December 2003, 32 children (mean age 7.2 +/- 5.4 years; 4 months - 15.9 years) underwent mechanical mitral (MVR, n = 17), aortic (AVR, n = 13) or double valve replacement (DVR, n = 2) with St. Jude Medical valves. Twenty-two patients (69 %) had undergone previous cardiac surgery. Anticoagulation self-management was used since 1995. RESULTS: The operative mortality was 3.1 %. Perioperative complications were complete heart block (n = 5), ventricular fibrillation (n = 1) and myocardial infarction (n = 1) and were exclusively related to patients with MVR. Mean calculated valve size ratio (geometric prosthesis orifice area/normal valve size area) was 1.72 (1.07 - 2.85) for AVR and 1.4 (0.88 - 3.12) for MVR. Mean follow-up was 9.1 +/- 6.6 years (range 0.4 - 23.2 years, cumulative 283 patient-years). There were two late deaths in patients with MVR. Actuarial survival after 10 years was 93.8 %. Late complications were endocarditis (n = 2), minor hemorrhagic event (n = 1), and stroke (n = 1). Anticoagulation self-management is well accepted by all patients/parents. Overall 10-year freedom from any anticoagulation-related adverse event with phenprocoumon was 89.1 % (1.2 %/patient year). Nine patients required reoperations: redo-MVR (outgrowth of prostheses (n = 3), pannus overgrowth (n = 2), closure of paravalvular leak after AVR (n = 2), partial aortic valve thrombosis (n = 1) and redo-DVR (n = 1 for endocarditis). Freedom from reoperation after 10 years was 80.9 %. CONCLUSIONS: Mechanical valve prostheses are a valuable option for left-sided heart valve replacement in pediatric patients. Perioperative morbidity was exclusively related to patients with MVR. Oversizing was often possible to avoid early reoperation for outgrowth. The operative mortality and long-term morbidity are acceptable. Anticoagulation self-management is safe and well accepted. PMID- 17410501 TI - A risk score to predict 30-day mortality in patients with intra-aortic balloon pump implantation. AB - BACKGROUND: Several risk factors for early mortality after intra-aortic balloon pump (IABP) implantation have been described. METHODS: We performed a retrospective analysis in 120 patients receiving peri- or postoperative IABP support. Univariate and multivariate analyses were carried out to assess variables potentially influencing 30-day mortality in order to develop a risk score for the prognosis of survival and for the decision on optimal patient specific treatment. RESULTS: The 4 parameters (mean arterial pressure, adrenaline dose, central venous pressure, and blood lactate concentrations) at 6 hours of IABP use were independently related to 30-day mortality. They were used to develop a risk score (0 - 4 points). With this score, patients who scored 3 or 4 points had no probability of surviving, whereas patients with a score of zero had a probability of 84.4 %. A prospectively screened cohort of 145 patients confirmed the reliability of our risk score. CONCLUSIONS: Our data demonstrate that a score can predict 30-day mortality in patients with IABP implantation. Such a score can be useful to find out whether or not ECMO/VAD implantation is necessary. PMID- 17410502 TI - Serum S100B levels in patients after cardiac surgery: possible sources of contamination. AB - BACKGROUND: S100B protein is considered to be a potential marker of brain damage. The aim of our study was to determine the contamination effect of retransfused blood on the serum S100B concentrations in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) and to differentiate between this simple contamination effect and its possible enhancement by haemolysis. METHODS: The first part of the study was performed in a group of 10 patients scheduled for coronary artery bypass grafting. Baseline S100B level was determined in a blood sample drawn from the radial artery before skin incision. After performing the distal anastomosis, additional blood samples were drawn from 1) the radial artery, 2) the aortic root catheter, 3) the pericardial space, and 4) CPB suction. To study the possible haemolytic effect on serum S100B levels, a second group of 23 patients was studied. S100B concentrations were determined in samples drawn simultaneously from the radial artery and bypass circuit after the end of CPB. Further samples from the retransfusion blood bag were analysed after one, two and three hours. RESULTS: Blood samples from the pericardial space and CPB suction exhibited significantly higher levels of S100B than the samples drawn from the peripheral artery and aortic root catheter in the first group of patients. No significant differences between the S100B levels in the peripheral blood and aortic root catheter were detected. In the second group, S100B was significantly elevated in the samples taken from the retransfusion blood bag in comparison with peripheral blood. S100B levels remained stable during the whole follow-up period. CONCLUSION: The results of our study show increased serum S100B levels caused by contamination originating in the mediastinal tissues. Storage of blood in the retransfusion bag and haemolysis can be excluded as sources of contamination. The role of S100B in perioperative monitoring of patients undergoing cardiac surgery remains to be established and should be confirmed by further studies using neuropsychological tests and imaging techniques. PMID- 17410503 TI - Arterial T-graft in coronary surgery: comparison of selective coronary angiography and multi-slice computed tomography. AB - BACKGROUND: The main aim of our study was to compare the results and value of angiography and multi-slice computed tomography (MSCT) after coronary artery bypass grafting using complex arterial conduit combinations. METHODS: Twenty-six patients underwent coronary surgery. In all patients, we utilized a T-graft (free arterial graft centrally implanted into the left internal thoracic artery IN SITU). Postoperative coronary angiography and MSCT were carried out prior to discharge. The results were interpreted separately by two different investigators. RESULTS: One hundred and thirteen distal anastomoses using 59 grafts for T-graft combinations were performed in 26 patients. A mean of 3.5 peripheral anastomoses was established per patient. Angiography showed a bypass graft patency rate of 94.6 % (56/59). In angiography, the patency rate of anastomoses was 96.5 % (109/113). All occlusions were identified in sequential anastomoses. The bypass patency rate measured by MSCT was 100 % and the patency rate of anastomoses 100 %. Based on these results, MSCT showed a sensitivity of 98.9 %. The specificity was 50.8 %, the positive predictive value was 94.6 % and the negative predictive value had a level of 99.9 %. CONCLUSION: MSCT is a simple, less invasive and useful method of monitoring results after coronary surgery including anastomosis and graft patency after complex arterial grafts. Its value may be restricted for sequential anastomoses and angiography should be preferentially used in these cases. PMID- 17410504 TI - Isolated lung perfusion with Paclitaxel and docetaxel: comparison of the early effects on pulmonary physiology. AB - BACKGROUND: This study was designed to compare the early effects of docetaxel and paclitaxel on pulmonary physiology after isolated lung perfusion. METHODS: Rats underwent isolated left lung perfusion with docetaxel in group 1 (n = 5), paclitaxel in group 2 (n = 5), and 0.9 %NaCl in the control group (n = 5). Ventilation pressures, compliance of the lungs, blood gas analysis and histopathological results were compared between the groups. RESULTS: In group 1 and group 2, the decrease in PaO (2) (p = 0.008) and increase in ventilation pressures were significantly higher than in the control group ( P = 0.016). In group 2, pCO (2) retention was higher compared to the docetaxel perfusion group ( P = 0.016). In the histochemical assessment, intra-alveolar hemorrhage and mononuclear cell infiltration were dense and perivascular edema was not present in group 1. In group 2, perivascular and intraalveolar edema were found to be dense. CONCLUSION: Perfusion by either of the chemotherapeutics resulted in an alteration of lung physiology in rat lungs. If isolated lung perfusion is administered using chemotherapeutics from the taxanes group, it is suggested that docetaxel could be the first choice for isolated lung perfusion. PMID- 17410505 TI - Experimental use of N-butyl cyanoacrylate tissue adhesive on lung parenchyma after pulmonary resection. AB - BACKGROUND: The aim of this study was to investigate the effectiveness of N-butyl cyanoacrylate tissue adhesive for the prevention of air leak together with the morphological changes to lung parenchyma. METHODS: Twelve New Zealand rabbits were used. The rabbits were ventilated with pressure-controlled ventilation during the experiment, beginning with a pressure level of 10 cm H (2)O. After a 2 x 2-cm pulmonary wedge resection, the resection surface was sealed with N-butyl cyanoacrylate and the pressure level was increased every five minutes in 5-cm H (2)O increments. The pressure level which caused an air leak from the resection surface was recorded. The morphological damage to the lung parenchyma was evaluated under light microscopy. RESULTS: The mean value of the pressure levels that caused air leak was 43.3 +/- 8.8 cm H (2)O. No tissue damage to lung parenchyma was recorded after histopathological examination. CONCLUSION: N-butyl cyanoacrylate was effective in preventing air leak from the pulmonary resection surface even with high airway pressure levels. It could be used as an aid for pulmonary resection lines or to control the air leak from pulmonary parenchyma. PMID- 17410506 TI - Transcutaneous electric nerve stimulation for the treatment of postthoracotomy pain: a randomized prospective study. AB - BACKGROUND: Insufficient relief of postthoracotomy pain is a major cause of increased rates of postoperative complications including inadequate coughing, mucous plugging, hypoxia, compromised ventilation or even bacterial lung infection. We aimed to assess the efficacy of transcutaneous electric nerve stimulation (TENS) in patients with postthoracotomy pain. METHODS: Forty patients scheduled to undergo posterolateral thoracotomy were randomly allocated to receive either TENS or patient-controlled intravenous morphine. Postoperative pain was evaluated using a visual analogue scale (VAS) and the Prince Henry pain scale. Pulmonary function was evaluated and an intergroup comparison was done. RESULTS: On the first three days following surgery, the VAS intensity of the TENS group did not differ significantly from that of the morphine group ( P > 0.05), and on the first two days following thoracotomy, the Prince Henry scale of the TENS group was not statistically significantly different. However, the VAS intensity was significantly lower than that of the control group on the fourth ( P = 0.044), fifth ( P = 0.016), sixth ( P = 0.009), seventh ( P = 0.008), eighth ( P = 0.004), ninth ( P = 0.002), tenth ( P = 0.001), fifteenth ( P = 0.002), thirtieth ( P < 0.001), forty-fifth ( P < 0.001) and sixtieth ( P < 0.001) days. The Prince Henry scale of the TENS group was found to be significantly diminished from the 3rd to the 60th day. TENS significantly reduced the analgesic requirements from day 5 to 60 ( P < 0.01). No noticeable side effect was observed in the TENS group during the study period. CONCLUSION: This study demonstrated that TENS provided a better pain relief and comfort compared to PCA from the fourth postoperative day onwards, and this pain-reducing effect continued for at least two months postoperatively. PMID- 17410507 TI - Primary malignant fibrous histiocytoma of the lung. AB - BACKGROUND: Malignant fibrous histiocytoma (MFH) is the most common soft tissue sarcoma in adults. Its appearance as a primary lung tumor is extremely rare. The cell origin of MFH remains controversial. The treatment of choice for MFH is surgical resection, while the role of chemo- and radiotherapy remains unclear. METHODS: A retrospective analysis of 5 patients operated on for primary MFH in the Department of Thoracic Surgery of the Medical University in Gdansk between 1990 and 2000 was performed. RESULTS: Out of approximately 2000 patients operated on for primary malignant lung tumors, five (0.25 %) had MFH. The mean age of the 4 men and 1 woman was 62 years. In all cases radical resection was performed without adjuvant chemo- or radiotherapy. Four patients died within 2 - 7 months after the operation, three of them from distant metastases. The follow-up of one patient is not available. One patient is alive 11 years after the operation. CONCLUSION: Although surgical resection of MFH is the treatment of choice in MFH, the results are unsatisfactory. PMID- 17410508 TI - The prognostic importance of trauma scoring systems for blunt thoracic trauma. AB - BACKGROUND: Early identification and aggressive management of blunt thoracic trauma are essential to reduce the significant rates of morbidity and mortality. The aim of this study was to evaluate the independent predictive value of 5 different trauma scoring systems (Revised Trauma Score [RTS], Trauma and Injury Severity Score [TRISS], Injury Severity Score [ISS], Lung Injury Scale [LIS], and Chest Wall Injury Scale [CWIS]) with respect to prognostic factors such as tube thoracostomy duration, the need for mechanical support and thoracotomy, the length of hospital and ICU stay, morbid conditions, and deaths of patients with blunt thoracic trauma. METHODS: The records of 152 patients with blunt thoracic trauma were reviewed and data consisting of the patients' age and gender, blood pressure and respiratory rate on admission, the extent of chest wall and intrathoracic injury, types of associated injuries, Glasgow Coma Scale (GCS) scores, the need for mechanical support and thoracotomy, tube thoracostomy duration, length of hospital and ICU stay, morbid conditions, and deaths were collected. The relations between the trauma scoring systems and prognostic factors were evaluated by multivariate analysis. RESULTS: The analysis showed that only TRISS was an independent predictor of mortality and only LIS was an independent predictor of morbidity, the need for thoracotomy, and tube thoracostomy duration. TRISS and LIS were independent predictors of the length of ICU stay. ISS, CWIS, and LIS were independent predictors of the need for mechanical support. RTS, TRISS, ISS and LIS were independent predictors of the length of hospital stay. CONCLUSIONS: The LIS grade appeared to correlate with the severity of blunt thoracic injury and was found to be the most useful scoring system in predicting the outcomes of these patients. PMID- 17410509 TI - Esophageal stent placement for the palliation of dysphagia in lung cancer. AB - BACKGROUND: The aim of this study was to evaluate the experience with expandable metal stents for the palliation of malignant dysphagia caused by lung cancer. METHODS: Between August 2002 and April 2006, we deployed esophageal metal stents in 14 patients with esophageal involvement from lung cancer. The mean age of the patients was 61.4 years, and 12 were male patients. Expandable metal stents were inserted under fluoroscopic control over a guide-wire with the patient under conscious sedation. RESULTS: Four patients had concomitant tracheoesophageal fistula. Eleven patients had received previous chemotherapy or radiotherapy and two had had a pneumonectomy. Immediate improvement after esophageal stent replacement was seen in all patients. The fistula was sealed off in all. All the patients remained asymptomatic during follow-up. All patients died, with mean survival of 10 weeks for patients with fistula and 8.3 months in the patients without fistula. CONCLUSION: The poor quality of life associated with malignant dysphagia and tracheoesophageal fistula in lung cancer patients can be significantly improved by a non-surgical intervention such as the deployment of expandable metal stents. PMID- 17410510 TI - Bilateral surgical resection in pulmonary epitheloid hemangioendothelioma. AB - Epitheloid hemangioendothelioma is a vascular tumour with an epitheloid appearance, originating from endothelial cells. Although it is a slow growing tumour, extensive pulmonary involvement, intrathoracic spread, and systemic spread have been documented. We present a case of epitheloid hemangioendothelioma of the lung in a patient with an initial diagnosis made by transthoracic biopsy. The prognosis is unpredictable, with life expectancy ranging from 1 to 20 years. There is no single effective treatment, though spontaneous regression and response to chemotherapy and interferon are reported. Our patient underwent pulmonary lobectomy of the right lower lobe and pulmonary wedge resection of the nodule located in the left lower lobe. PMID- 17410511 TI - Laser-assisted resection of a giant pulmonary chondrohamartoma--a case report. AB - A rare case of a giant pulmonary chondromatous hamartoma (15 cm, 1350 g) resected by a new laser system (Nd:YAG, 1318 nm, 40 W) is presented. The laser management of a hamartoma resection--the largest reported to date in the literature--is presented here. PMID- 17410512 TI - Finding the limit between station 2 and station 4 during right-sided thoracotomy. AB - In mediastinal dissection through a right thoracotomy, the definition of station 2 is arbitrary because no anatomical landmark indicates the line drawn tangentially to the upper margin of the aortic arch. We have developed a technique to localize it by evaluating the distance between the upper aortic arch and the azygos vein on a CT scan. This distance located intraoperatively above the azygos vein permits the surgeon to draw an imaginary line parallel to the azygos vein, which we consider to be the limit between station 2 and station 4. To verify the precision of the technique, an 8-mm clip was positioned at the intersection between the imaginary line dividing station 2 and station 4 and the superior vena cava in 38 consecutive right-sided lateral muscle-sparing thoracotomies. The definition of the station 2/4 limit was defined as "excellent" if the upper aortic arch line crossed the clips, "good" if clips were 1000 microM. The highest cytotoxicity towards AGS cells was observed for the 2 bromophenyl- and 2-hydroxy-5-chlorophenylamides in both diterpene series, with IC50 values in the range of 14-34 microM towards AGS cells and 10-37 microM towards fibroblasts, respectively. PMID- 17410518 TI - Supportive care of rural women with breast cancer in Tasmania, Australia: changing needs over time. AB - Providing for patients' psychosocial needs is a potential means of minimising cancer morbidity. Needs assessments can guide responsive patient-centred care. A longitudinal survey of women with early breast cancer consulting a breast nurse in a primarily rural state of Australia was undertaken to measure unmet supportive care needs, identify changes in unmet needs across time and compare results with previous studies. Needs assessments were completed with the Supportive Care Needs Survey (SCNS) at 1 month (n = 74) and 3 months (n = 83) post-diagnosis. Access to services was also examined. High levels of psychological and health system and information needs were identified at 1 month post-diagnosis, but these decreased significantly at 3 months post-diagnosis. Sexuality domain needs increased significantly during the same time. Compared to a previous SCNS study of rural women with breast cancer, unmet supportive care needs in this study were significantly lower than previously reported, although assistance with energy levels (i.e. fatigue) continues to be an unmet need for women with breast cancer. Results suggest there have been positive cultural changes within healthcare systems, specifically in better informing patients and providing support. Although services appear to be more responsive to breast cancer patients, significant unmet needs still exist. PMID- 17410519 TI - Biomolecular recognition of glycosylated beta(3)-peptides by GalNAc specific lectins. AB - The molecular recognition of a novel kind of hybrid conjugates, composed of artificial biomimetic beta-peptide oligomers with an O-linked natural N-acetyl galactosamine (the Tn-antigen) residue, by four different GalNAc specific lectins was investigated using surface plasmon biosensor technology. The influence of the peptide and the glycosyl moiety on the recognition was studied using two glycosylated beta(3)-heptapeptides, a glycosylated alpha-heptapeptide, two beta amino acid containing dipeptides, and monomeric alphaGalNAc-O-Thr. Although all four lectins displayed a decreased affinity for the carbohydrate residue when attached to a peptide, as compared to the monomeric Tn-antigen, the peptide part was found to have distinct effects on the binding kinetics-indicating that varying degrees of protein-peptide interactions occurred in the recognition process. Likewise, the lectins did not discriminate between beta(3)-peptides and the alpha-peptide, but the beta-linkage of the galactose had a detrimental effect for at least two of the lectins. PMID- 17410520 TI - Adaptive style and differences in parent and child report of health-related quality of life in children with cancer. AB - The objective of this study was to examine the self-reported health-related quality of life (HRQL) of children with cancer, and the consistency between child and parent reports of child HRQL, as a function of the child's adaptive style. Participants included 199 children with cancer, 108 healthy children, and their parents. Children completed self-report measures of HRQL and adaptive style. Measures of adaptive style were used to categorize children as high anxious, low anxious, defensive high anxious or repressor. Parents completed measures reporting their children's HRQL. Adaptive style was a significant predictor of child-reported HRQL, particularly on the psychosocial scales, with children identified as repressors reporting the best HRQL. Adaptive style was also predictive of discrepancies between parent and child report of child HRQL. Repressor and low anxious children reported better HRQL than did their parents, while high anxious children reported poorer HRQL, regardless of health status. Adaptive style is a significant determinant of self-reported HRQL in children, particularly in psychosocial domains, while health status (i.e. cancer patient vs healthy control) is predictive only of physical health domains. Researchers and clinicians should be aware of the impact of child adaptive style when assessing HRQL outcomes using self- or parent report. PMID- 17410521 TI - The construction of standard gamble utilities. AB - Health effects for cost-effectiveness analysis are best measured in life years, with quality of life in each life year expressed in terms of utilities. The standard gamble (SG) has been the gold standard for utility measurement. However, the biases of probability weighting, loss aversion, and scale compatibility have an inconclusive effect on SG utilities. We determined their effect on SG utilities using qualitative data to assess the reference point and the focus of attention. While thinking aloud, 45 healthy respondents provided SG utilities for six rheumatoid arthritis health states. Reference points, goals, and focuses of attention were coded. To assess the effect of scale compatibility, correlations were assessed between focus of attention and mean utility. The certain outcome served most frequently as reference point, the SG was perceived as a mixed gamble. Goals were mostly mentioned with respect to this outcome. Scale compatibility led to a significant upward bias in utilities; attention lay relatively more on the low outcome and this was positively correlated with mean utility. SG utilities should be corrected for loss aversion and probability weighting with the mixed correction formula proposed by prospect theory. Scale compatibility will likely still bias SG utilities, calling for research on a correction. PMID- 17410522 TI - Vascular endothelial growth factor and psychosocial factors in colorectal cancer. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) and psychosocial factors have both been shown to have independent prognostic value in colorectal cancer. Recently, an association between VEGF and psychosocial factors has been reported in patients with ovarian cancer. METHODS: A consecutive series of eligible patients undergoing elective resection for colorectal cancer were invited to participate. Standardized measures of various aspects of quality of life (QoL) were administered 5-12 days before surgery, and again 6-8 weeks after the operation, blood samples were obtained at the same times. Solid phase ELISA was used for VEGF-A analysis using serum samples. Correlations and multiple regression analysis were used to examine the relationship between VEGF and psychosocial factors. RESULTS: One hundred and four patients with colorectal cancer were recruited. Seventy (67%) were male and the mean age was 67.6 years.A significant positive correlation was found between preoperative VEGF level and preoperative depression (r=0.227, p=0.03). Preoperative VEGF was negatively correlated with preoperative cancer-related concerns (r=-0.273, p=0.009) and positive affectivity (r=-0.219, p=0.05). Linear regression using TNM stage as a covariate showed that cancer-related concerns were an independent predictor of preoperative VEGF levels (p=0.02). Preoperative cancer-related concerns and global QoL were negatively correlated with postoperative VEGF (r=-0.381, p=0.001, r=-0.264, p=0.005), whereas preoperative depression correlated positively with postoperative VEGF (r=0.333, p=0.003). Linear regression analysis showed that preoperative HADS depression (p=0.005) and cancer-related concerns (p=0.002) were independent predictors of postoperative VEGF levels. Postoperative VEGF was significantly correlated with postoperative anxiety (r=0.249, p=0.02), depression (r=0.289, p=0.01), and functional well-being (r=-0.242, p=0.03). Linear regression analysis showed that postoperative anxiety (p=0.048), depression (p=0.013) and functional well-being (p=0.046) independently predicted postoperative VEGF-A levels. CONCLUSION: Various psychosocial factors, particularly cancer-related concerns and depression, appear to be related to preoperative and postoperative VEGF level in patients with newly diagnosed colorectal cancer. However, the clinical significance of these findings needs to be addressed in longitudinal follow-up studies of recurrence and survival. Future prognostic studies involving VEGF and related cytokines should assess psychosocial variables at various time points and include these in a multivariate analysis of outcomes. PMID- 17410523 TI - IgVH gene mutation status and genomic imbalances in chronic lymphocytic leukaemia with increased prolymphocytes (CLL/PL). AB - Chronic lymphocytic leukaemia (CLL) with increased prolymphocytes (CLL/PL) has been defined by the World Health Organization (WHO) classification and considered as a progressive and clinically aggressive variant of CLL. To further characterize the biological features of this disease, we performed IgVH gene mutational status, FISH and high-resolution comparative genomic hybridization (HR CGH) analysis in 17 cases of CLL/PL. All CLL/PL utilized members of VH1, VH3 and VH4 families, with the highest prevalence of the VH1-69 gene. In all but one cases analyzed, the VH genes were unmutated. The FISH and HR-CGH analyses showed frequent occurrence of trisomy 12, del(11)(q23), del(17)(p13) and genetic imbalances, but recurrent genetic lesion characteristic for CLL/PL was not found. The follow-up HR-CGH analysis of two cases showed that increase of prolymphocytes in the course of CLL or CLL/PL are associated with clonal evolution and selection of the tumour clone. In conclusion, this study suggests that CLL/PL is a relatively homogeneous disease regarding VH gene mutation, but heterogeneous regarding genetic lesions. The heterogeneity and high number of genomic imbalances found in CLL/PL suggest that a genome-wide instability of the neoplastic cells may play a role in the development of the disease. PMID- 17410524 TI - In Vitro and In Vivo pharmacokinetic studies of bulaquine (analogue of primaquine), a novel antirelapse antimalarial, in rat, rabbit and monkey- highlighting species similarities and differences. AB - Bulaquine (BQ) is a potent antirelapse antimalarial developed by CDRI, India. Bulaquine was rapidly absorbed in rats and rabbits with no distinct absorption phase while in monkeys a variable irregular absorption profile was observed. BQ was extensively converted to primaquine (PQ) after oral administration and the conversion was maximum in rats and minimum in rabbits, which is possibly due to the species difference. Clearance was higher in rats (3.2 l/h/kg) than in rabbits and monkeys (1.2 l/h/kg) and it was found be negligibly excreted in rat urine and feces. The elimination half-life in rats and rabbits was comparable after both oral and i.v. administration ( approximately 1.2 h). In all three species, PQ was resident in the body for a period longer than BQ. PQ, being the major active metabolite of BQ, might be responsible for the extended therapeutic effect of BQ. The oral bioavailability of BQ was 3.12%, 5.3% and 12% in rats, rabbits and monkeys, respectively, which could be mainly due to the high instability of BQ at acidic pH as demonstrated from a simulated gastric fluid stability study. Protein binding in various species was in the range 50-65% while the partition coefficient between RBCs and plasma (K(rbc/pl)) was between 0.75 and 1, indicating significant RBC uptake. PMID- 17410525 TI - Progression to impaired glucose regulation, diabetes and metabolic syndrome in Chinese women with a past history of gestational diabetes. AB - BACKGROUND: To examine the risk of developing impaired glucose regulation (IGR), diabetes mellitus (DM) and metabolic syndrome (MetS) in Chinese women with history of gestational diabetes. METHODS: 203 Chinese women enrolled in a previous study were followed up at a median of 8 (range 7-10) years of whom 136 had normal glucose tolerance (NGT) and 68 had gestational diabetes mellitus (GDM) and gestational impaired glucose tolerance (GIGT). RESULTS: In women with a history of gestational diabetes (n = 4), GIGT (n = 63) and NGT (n = 136), 2 (50%), 19 (30.2%) and 21 (15.4%) developed IGR while 2 (50%), 4 (6.3%), 3 (2.2%) developed DM respectively. Most women developed IGR (86%, n = 36) or DM (78%, n = 7) were undiagnosed. MetS occurred in 16 (7.9%) women with similar rates between those with and those without a history of gestational diabetes (7.5% vs 8.1%; p = 0.85). History of gestational diabetes [OR: 3.8 (95% CI 1.9-7.8)] and body mass index (BMI) >/= 23 kg/m(2) [OR: 3.4 (95% CI 1.7-6.8)] at first antenatal visit were predictors for IGR or DM. Family history of DM [OR: 5.0 (95% CI 1.5-16.4)] and BMI >/= 23 kg/m(2) [OR: 28.3 (95% CI 3.6-223)] at first antenatal visit were predictors for MetS. CONCLUSIONS: Chinese women with a history of gestational diabetes have a high risk of IGR or DM. Overweight at the first antenatal visit is a common risk factor for IGR, DM and MetS. A prior history of gestational diabetes was predictive of IGR and DM while a positive family history of DM was predictive of MetS. PMID- 17410526 TI - The use and perceived benefit of complementary and alternative medicine among Californians with cancer. AB - Population-based data on complementary and alternative medicine (CAM) use among cancer sufferers is lacking. In a telephone survey representative of California households (response rate = 68.9%, N = 1845) those who reported a diagnosis of cancer (excluding non-melanoma skin cancers) were asked about CAM use. CAM use is substantial, although with few exceptions, it is approximately that found among those with non-malignant chronic conditions. Those with cancer are more likely to report praying for their health, using support groups, and taking multiple dietary supplements. They are less apt to use CAM providers or special diets. Socio-demographic factors associated with CAM use vary by specific CAM modality. Site of the cancer was not associated with any particular CAM modality. The use of CAM therapies specifically promoted as cancer therapies was not common, especially among those diagnosed recently. The use of CAM providers and mind-body techniques specifically for the purpose of treating cancer is unusual (<10%), while special diets are more frequently employed for the purpose of treatment and/or prevention of the cancer itself. A clear majority of those who do use CAM for treating cancer report at least some benefit from the treatment, and are likely to inform their physicians of such use. PMID- 17410527 TI - In situ kinetic modelling of intestinal efflux in rats: functional characterization of segmental differences and correlation with in vitro results. AB - The objective was to devise and apply a novel modelling approach to combine segmental in situ rat perfusion data and in vitro cell culture data, in order to elucidate the contribution of efflux in drug absorption kinetics. The fluoroquinolone CNV97100 was used as a model P-gp substrate. In situ intestinal perfusion was performed in rat duodenum, jejunum, ileum and colon to measure the influence of P-gp expression on efflux. Inhibition studies of CNV97100 were performed in the presence of verapamil, quinidine, cyclosporin A and p aminohippuric acid. Absorption/efflux parameters were modelled simultaneously, using data from both in situ studies as well as in vitro studies. The maximal efflux velocity was modelled as a baseline value, corrected for each segment based on the expression level. CNV97100 passive diffusional permeability (P(diff)) and its affinity for the efflux carrier (K(m)) were assumed to be the same in all segments. The results indicate the new approach to combine in situ data and in vitro data succeed in yielding a unified, quantitative model for absorption/efflux. The model incorporated a quantitative relationship between P gp expression level and the efflux functionality, both across in situ and in vitro systems, as well across different intestinal segments in the in situ studies. Permeability values decreased from duodenum to ileum in accordance with the increasing P-gp expression levels in rat intestine. The developed model reflects a strong correlation between in vitro and in situ results, including intrinsic differences in surface area. The successful application of a model approach to combine absorption data from two different experimental systems holds promise for future efforts to predict absorption results from one system to a second system. PMID- 17410528 TI - Factors associated with breast cancer-specific distress in younger women participating in a family history mammography screening programme. AB - This multi-centre study examined factors associated with breast cancer-specific distress in 2321 women under 50 who are on a mammographic screening programme on account of their family history. Women were recruited from 21 UK centres, and completed a questionnaire one month before their screening appointment. The transactional theory of stress, appraisal, and coping provided the theoretical framework for the study. Factors measured included screening history, family history, perceived risk, cognitive appraisals, coping, optimism, and cancer worry. The findings indicate that the majority of women appraise their family history as being relevant and somewhat threatening to personal well-being, but something they can deal with emotionally. Acceptance was the most commonly used coping strategy. Hierarchical regression analysis identified that the factors most significantly associated with distress were an appraisal of high relevance and threat, increased risk perception, low dispositional optimism, and the use of both avoidant and task-orientated coping strategies. Women with children and those with relatives who have died from breast cancer were also more distressed. To conclude, most women appraised their situation positively but there is a potential profile of risk factors which may help clinicians identify those women who need extra psychological support as they progress through screening. PMID- 17410529 TI - Interpreting analyses of continuous covariates in affected sibling pair linkage studies. AB - Datasets collected for linkage analyses of complex human diseases often include a number of clinical or environmental covariates. In this study, we evaluated the performance of three linkage analysis methods when the relationship between continuous covariates and disease risk or linkage heterogeneity was modeled in three different ways: (1) The covariate distribution is determined by a quantitative trait locus (QTL), which contributes indirectly to the disease risk; (2) the covariate is not genetically determined, but influences the disease risk through statistical interaction with a disease susceptibility locus; (3) the covariate distribution differs in families linked or unlinked to a particular disease susceptibility locus. We analyzed simulated datasets with a regression based QTL analysis, a nonparametric analysis of the binary affection status, and the ordered subset analysis (OSA). We found that a significant OSA result may be due to a gene that influences variability in the population distribution of a continuous disease risk factor. Conversely, a regression-based QTL analysis may detect the presence of gene-environment (GxE) interaction in a sample of primarily affected individuals. The contribution of unaffected siblings and the size of baseline lod scores may help distinguish between QTL and GxE models. As illustrated by a linkage study of multiplex families with age-related macular degeneration, our findings assist in the interpretation of analysis results in real datasets. They suggest that the side-by-side evaluation of OSA and QTL results may provide important information about the relationship of measured covariates with either disease risk or linkage heterogeneity. PMID- 17410530 TI - Affected relative pairs and simultaneous search for two-locus linkage in the presence of epistasis. AB - It is commonly believed that multiple interacting genes increase the susceptibility of genetically complex diseases, yet few linkage analyses of human diseases scan for more than one locus at a time. To overcome some of the statistical and computational limitations of a simultaneous search for two disease susceptibility loci in the presence of epistasis, we developed new score statistics to simultaneously scan for two disease susceptibility loci in pedigree data. These model-free score statistics are based on developments for model-free maximum lod scores, which in turn are based on variance components for indicators of disease status. To overcome reduced power caused by many parameters in the general two-locus model, we impose constraints on ratios of variance components, much like those used for robust single-locus linkage statistics (e.g., minimax constraints). The resulting three-degree of freedom score statistic, constrained as a one-sided multivariate test, can be computed rapidly, making simultaneous search feasible for human genetic linkage studies. Furthermore, using recent developments to rapidly compute simulation P-values for score statistics, it is feasible to empirically evaluate the statistical significance of the proposed score statistics. Application of these methods to two large studies of the genetic linkage of prostate cancer illuminates their strengths and limitations. The results provide weak suggestions for linkage of several pairs of chromosomal regions (chromosome pairs 1-21, 3-13, 5-9, and 14-19), all of which showed stronger linkage signals when the score statistics accounted for epistasis. These novel score statistics should prove useful for linkage studies of other complex human diseases. PMID- 17410531 TI - Antioxidants modulate the IL-6 induced inhibition of negative acute-phase protein secretion in HepG2 cells. AB - Despite increasing evidence on the potential of dietary antioxidants in modulating the etiology of certain chronic diseases such as cancer and cardiovascular diseases, little is known about their beneficial role in acute phase responses and inflammatory diseases. From this viewpoint the aim of this study was to investigate the effect of selected dietary antioxidants in modulating the secretion of negative acute-phase proteins caused by interleukin-6 (IL-6) in HepG2 cells. Cells were first stimulated with a fixed dose of IL-6 for 24 h then incubated for a further 8 h with varying concentrations of eight antioxidants, alpha-lipoic acid (LA), (-)-epicatechin (EC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin (EGC), (-)-epigallocatechin gallate (EGCG), alpha-tocopherol (TOC), ascorbic acid (AA) and N-acetylcysteine (NAC). The culture supernatants were assayed for transthyretin (TTR) and retinol binding protein (RBP) using ELISA. The data revealed that IL-6 significantly reduced TTR and RBP secretion compared with the basal production. All tested antioxidants attenuate the reduction in TTR and RPB levels. The strongest effects were achieved with the highest concentration of each antioxidant. The order of effect were LA > EGCG > ECG > TOC > EGC > EC > NAC > AA. In conclusion, these results provide evidence that the dietary antioxidants can play a fundamental role in inflammatory processes. PMID- 17410532 TI - Multivariate prediction of the probability of recurrence in patients with carcinoma of the parotid gland. AB - BACKGROUND: Parotid gland carcinoma is an infrequent tumor, and series that report on these neoplasms are relatively scarce in the literature. The objective of the current study was to identify prognostic factors in patients with parotid gland carcinoma and to develop a method for defining the probability of recurrence. METHODS: Patients with parotid gland carcinoma who were treated at the authors' institution from January 1981 through December 2004 and who completed treatment constituted the study group. Disease-free survival was calculated by using the Kaplan-Meier method. Logistic regression analysis was employed to define the recurrence-associated prognostic factors. RESULTS: One hundred twenty-seven patients were included (64 men and 63 women); their mean age was 53 years. Mucoepidermoid carcinoma was diagnosed in 34.6% of patients, adenoid cystic was diagnosed in 15.7% of patients, adenocarcinoma was diagnosed in 14.3% of patients, and acinic cell carcinoma was diagnosed in 9.4% of patients. The median disease-free survival was 8.3 years (95% confidence interval [95% CI], 4.3-12.2 years). Logistic regression analysis confirmed tumor classification, facial nerve palsy, grade of tumor differentiation, patient age, and surgical margins as recurrence-associated factors (P < .00001). Using this model, 3 postoperative risk groups were defined-high-risk, intermediate-risk, and low-risk-that had recurrence frequencies of 71.4%, 43.1%, and 8.8%, respectively (P = .0001). The 5-year disease-free survival rates for these groups were 18.7%, 53.9%, and 99.9%, respectively (P = .00001). CONCLUSIONS: In this study, the authors identified several significant prognostic factors. Consequently, they have proposed a prognostic score categorization that allows for a straightforward calculation of the risk of recurrence for a given patient that may help to define therapeutic strategies, target patient counseling, and design future trials. PMID- 17410533 TI - Surveillance endoscopy does not improve survival for patients with local and regional stage colorectal cancer. AB - BACKGROUND: Endoscopic surveillance is recommended and widely practiced after definitive treatment for colorectal cancer, yet to the authors' knowledge there is little evidence supporting its benefit. The purpose of the current study was to estimate the impact of endoscopic surveillance on colorectal cancer-specific survival for persons with localized or regional colorectal cancer. The population included Medicare patients (age >or=65 years) who were diagnosed with local or regional stage colorectal cancer between 1986 and 1996. METHODS: The current study was a retrospective case-control study. Cases were defined as those individuals who died of colorectal cancer and controls were defined as those with colorectal cancer who did not die of colorectal cancer; controls were frequency matched to cases. Surveillance was defined as the use of colonoscopy, flexible sigmoidoscopy, or barium enema >or=6 months after diagnosis. Logistic regression was used to control for endoscopic procedure, race, comorbidity index at the time of diagnosis, and types of initial treatments after surgery. RESULTS: The analysis group contained 8130 cases (29%) and 20,079 controls (71%). The average time to first bowel surveillance for those with at least 1 surveillance examination was 15.9 months after the diagnosis (median, 13 months). In the regression analysis, surveillance endoscopy was not found to be associated with improved colorectal cancer-specific survival (odds ratio of 1.01; 95% confidence interval, 0.95-1.06 [P=0.85]). Setting the surveillance interval to 12 months and 15 months rather than 6 months after diagnosis did not appear to influence the results. CONCLUSIONS: Surveillance endoscopy does not appear to influence colorectal cancer-specific mortality in patients age >65 years who are diagnosed with localized or regional stage colorectal cancer. PMID- 17410534 TI - Comparison of gene expression data from human and mouse breast cancers: identification of a conserved breast tumor gene set. AB - The aim of our work was to establish a database for breast cancer gene expression data in order to compare human and mouse breast cancer. We identified human and mouse homologues genes and compared the expression profile of 24 human breast tumors with 6 WAP-SVT/t breast tumors (WAP-SVT/t animals, line 8). Our studies confirmed the heterogeneity in gene expression of human as well as mouse breast cancer cells. However, 63 genes were found to be differentially expressed (upregulated: 40; downregulated: 23 genes) in at least 75% of the breast tumors of both species. To differentiate between early and late events in tumor formation, we compared the 63 differentially expressed genes with a mouse data set obtained from hyperplastic mammary glands. This revealed that the majority of the early deregulated genes are cell proliferation specific. These early changes seem to be necessary although not sufficient for breast cancer formation. Late alterations concern mainly genes belonging to the category of cell communication and metabolism. Interestingly, most of the 63 conserved genes are commonly associated with tumorigenesis. PMID- 17410535 TI - Over-expression of parathyroid hormone Type 1 receptor confers an aggressive phenotype in osteosarcoma. AB - Osteosarcoma is the most common primary bone malignancy in children and is associated with rapid bone growth. Parathyroid hormone-related peptide (PTHrP) signaling via parathyroid hormone Type 1 receptor (PTHR1) is important for skeletal development and is involved in bone metastases in other tumors. The aim of this study was to investigate the status of PTHrP/PTHR1 and its possible role in osteosarcoma. In a preliminary screening, a higher level of PTHR1 mRNA, but not PTHrP, was found in 4 osteosarcoma xenografts as compared with 4 standard cell lines, or 5 patient derived cell lines (p < 0.05) using quantitative RT-PCR. It was therefore extended to 55 patient specimens, in which a significantly higher level of PTHR1 mRNA was detected in metastatic or relapsed samples than those from primary sites (p < 0.01). Cell behavior caused by PTHR1 overexpression was further studied in vitro using PTHR1 transfected HOS cell line as a model. Over-expression of PHTR1 resulted in increased proliferation, motility and Matrigel invasion without addition of exogenous PTHrP suggesting an autocrine effect. Importantly, the aggressiveness in PTHR1-expressing cells was completely reversed by RNAi mediated gene knockdown. In addition, PTHR1 over-expression led to delayed osteoblastic differentiation and upregulation of genes involved in extracellular matrix production, such as TGF-beta1 and connective tissue growth factor. When cocultured with bone marrow derived monocytes, PTHR1 transfected HOS cells induced a greater number of osteoclasts. This study suggests that PTHR1 over-expression may promote osteosarcoma progression by conferring a more aggressive phenotype, and forming a more favorable microenvironment. PMID- 17410536 TI - Mechanisms of synthetic serine protease inhibitor (FUT-175)-mediated cell death. AB - BACKGROUND: Constitutive activation of nuclear factor-kappaB (NF-kappaB) is a frequent molecular alteration in pancreatic cancer and a number of studies have suggested that constitutive NF-kappaB activity plays a key role in the aggressive behavior of this disease. In an attempt to identify an effective therapeutic agent for pancreatic cancer, the authors studied the role of FUT-175, a synthetic serine protease inhibitor, in the inhibition of NF-kappaB activation and the induction of apoptotic responses. METHODS: To examine the effect of FUT-175 on the inhibition of NF-kappaB and the induction of apoptosis in pancreatic cancer cell lines, Western and Northern blot analyses, electromobility shift (EMSA), luciferase reporter gene, DNA fragmentation, immunoprecipitation, in vitro kinase, small interfering RNA (siRNA), and chromatin immunoprecipitation (ChIP) assays were performed. RESULTS: In a time-dependent and dose-dependent manner, FUT-175 inhibited IkappaBalpha phosphorylation and NF-kappaB activation, thereby inhibiting the antiapoptotic activity of NF-kappaB. Simultaneously, FUT-175 up regulated the expression of tumor necrosis factor receptor-1 (TNFR1), which in turn activated the proapoptotic caspase-8 and Bid pathways and induced apoptosis in pancreatic cancer cells. FUT-175-induced activation of Fas-associated death domain (FADD) and caspase-8 was suppressed by RNA interference-mediated inhibition of TNFR1 expression. Furthermore, expression of the transcription factor PEA3 was up-regulated by FUT-175 and was involved in FUT-175-mediated TNFR1 expression. CONCLUSIONS: These results suggested a possible mechanism by which FUT-175 may disrupt interconnected signaling pathways by both suppressing the NF-kappaB antiapoptotic activity and inducing TNFR-mediated apoptosis. Supported by this unique function as a NF-kappaB inhibitor and apoptosis inducer, this well-established synthetic serine protease inhibitor with as-of-yet poorly understood mechanisms of actions appears to be a potentially therapeutic agent for pancreatic cancer. PMID- 17410537 TI - Effect of familial history and smoking on common cancer risks in Japan. AB - BACKGROUND: Inherited genetic predispositions are important risk factors for the development of cancer in general. To determine genetic susceptibility for 14 common cancers, a case-control study of the impact of a family history of cancer in first-degree relatives was conducted. The authors further evaluated the effect modification by habitual smoking with adjustment for other confounding environmental factors. METHODS: In total, 18,836 cancer cases and 28,125 age matched and sex-matched controls, confirmed as being free of cancer, were recruited. Odds ratios (ORs) and 95% confidence intervals were determined by multiple logistic regression analysis, including stratification by family history for 14 cancer sites and interactions with a smoking history. RESULTS: The associations between family history and risk of cancer were generally stronger at the same sites than across cancer sites. Risks to first-degree relatives at the same sites were found to be significantly elevated with 8 of 14 cancer sites; especially high ORs were found for prostate and thyroid cancers. Some across-site associations were observed; in particular, a reciprocal association between breast and prostate cancer was found. The interaction between family history and smoking history for breast cancer was found to be statistically significant. There was no statistical evidence for the interactions in other sites, but among subjects with a family history, the ORs were found to be higher in smokers compared with nonsmokers. CONCLUSIONS: The results of the current study support the hypothesis of a genetic susceptibility to cancers in family members. For breast cancer, the interaction between family history and smoking history was observed to be significant. PMID- 17410538 TI - Establishment of association of an Mg2+-dependent endonuclease with the rat liver nuclear matrix in cryonecrosis. AB - Previously, we characterized the endonucleolytic activity of the nuclear matrix prepared from rat liver cryopreserved in liquid nitrogen. The enzymic activity was attributed to a 23 kDa, Mg(2+)-dependent and sequence non-specific endonuclease (p23) stably associated with the nuclear matrix. Here we show that p23 was absent from the nuclear matrix prepared from fresh liver. Instead, both ex vivo (cryopreservation), as well as in vivo-induced necrosis by repeated freezing/thawing of liver tissue in an anaesthetized rat, promoted the activation and translocation of p23 to the nuclear matrix. Considering that ex vivo and in vivo freezing/thawing of the liver were accompanied by morphological (nuclear compaction) and biochemical events (increased LDH activity, disorderly genomic DNA degradation, absence of lamin proteolysis, appearance of 62 and 50 kDa necrotic cleavage products of PARP-1) commonly observed during necrosis, and because the association of p23 with the nuclear matrix was saturable, reflecting the existence of a limited number of distinct high affinity sites on the nuclear matrix for p23, we concluded that the activation of the nuclear matrix-associated endonuclease p23 is a feature of liver cryonecrosis. Although cryonecrosis represents a typical example of acute cell damage, our results suggest that it is realized by ordered molecular events. PMID- 17410539 TI - SELDI-TOF MS versus prostate specific antigen analysis of prospective plasma samples in a nested case-control study of prostate cancer. AB - There is an urgent need for better biomarkers for detection of clinically significant prostate cancer (PCa). Recent studies suggest that surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) analysis of serum may provide diagnostic information. The aim of this study was to investigate if PCa cases could be identified by applying predefined SELDI-TOF analysis conditions on prospectively, uniformly collected plasma samples from PCa cases and matched controls. Prospective samples from 387 incident PCa cases and an equal number of controls, matched for age and time for recruitment, were analyzed by SELDI-TOF MS (IMAC30/Cu chip) and multivariate classification analysis. Prospective prostate specific antigen levels were subjected to ROC curve analysis giving an AUC of 0.87 for the total cohort with a median lag time between blood sampling and diagnosis of 6.1 years. No markers were found by SELDI TOF MS that significantly discriminated between cases and controls in the total cohort or in subanalysis of cases with less than 2 years between blood donation and diagnosis (n = 42). Cases with aggressive disease at the time of diagnosis who gave blood less than 4 years prior to diagnosis (n = 23) could however be separated from their controls (sensitivity 70%, specificity 83%) by a model based on SELDI-TOF MS and OPLS-DA data analysis. We were thus not able to confirm previous results with SELDI-TOF MS identifying men with PCa from healthy individuals, but we report an optimal experimental set-up for verification of markers for early detection of cancer in prospectively collected samples. PMID- 17410540 TI - Protein kinase C activity in mouse eggs regulates gamete membrane interaction. AB - Gamete membrane interaction is critical to initiate the development of a new organism. The signaling pathways governing this event, however, are poorly understood. In this report, we provide the first evidence that protein kinase C activity in mouse eggs plays a crucial role in the regulation of this process. Stimulating PKC activity in mouse eggs by phorbol 12-myristate 13-acetate (PMA) drastically inhibited the egg's membrane ability to bind and fuse with sperm. Surprisingly, this significant reduction of gamete membrane interaction was also observed in eggs treated with the PKC inhibitors staurosporine and calphostin c. In further analysis, we found that while no change of egg actin cytoskeleton was detected after either PMA or calphostin c treatment, the structural morphology of egg surface microvilli was severely altered in the PMA-treated eggs, but not in the calphostin c-treated eggs. Moreover, sperm, which bound but did not fuse with the eggs treated with the anti-CD9 antibody KMC8, were liberated from the egg membrane after PMA, but not calphostin c, treatment. Taken together, these results suggest that egg PKC may be precisely balanced to regulate gamete membrane interaction in a biphasic mode, and this biphasic regulation is executed through two different mechanisms. PMID- 17410541 TI - An alternative Alcian Blue dye variant for the evaluation of fetal cartilage. AB - BACKGROUND: The most comprehensive evaluation of vertebrate skeletal development involves the use of Alizarin Red S dye to stain ossified bone and various other dyes to stain cartilage. The dye used most widely to stain fetal cartilage in rodents and rabbits is Alcian Blue 8GX. However, the global supply of this specific dye has been exhausted. Several forms of the dye marketed as Alcian Blue 8GX are now available, although they are not synthesized via the original 8GX manufacturing process. METHODS: One new Alcian Blue 8GX form and two Alcian Blue dye variants were evaluated in rats and rabbits using standard staining procedures. The staining quality of these dyes were evaluated relative to the original form of Alcian Blue 8GX based on cartilage uptake of the dye, clarity of the cartilaginous components, staining intensity of the dye, and overall readability of the specimens under stereomicroscopic evaluation. RESULTS: Staining with the newer form of Alcian Blue 8GX resulted in poor staining quality. The Alcian Blue-Pyridine variant performed well, although staining intensity was less than optimal. The Alcian Blue-Tetrakis variant provided staining characteristics that were most similar to the original form of Alcian Blue 8GX. CONCLUSIONS: Alcian Blue-Tetrakis was markedly better in its ability to stain fetal cartilage than the newer form of Alcian Blue 8GX. PMID- 17410543 TI - Morphological changes and spatial regulation of diacylglycerol kinase-zeta, syntrophins, and Rac1 during myoblast fusion. AB - The fusion of mononuclear myoblasts into multinucleated myofibers is essential for the formation and growth of skeletal muscle. Myoblast fusion follows a well defined sequence of cellular events, from initial recognition and adhesion, to alignment, and finally plasma membrane fusion. These processes depend upon coordinated remodeling of the actin cytoskeleton. Our recent studies suggest diacylglycerol kinase-zeta (DGK-zeta), an enzyme that metabolizes diacylglycerol to yield phosphatidic acid, plays an important role in actin reorganization. Here, we investigated whether DGK-zeta has a role in the fusion of cultured C2C12 myoblasts. We show that DGK-zeta and syntrophins, scaffold proteins of the dystrophin glycoprotein complex that bind directly to DGK-zeta, are spatially regulated during fusion. Both proteins accumulated with the GTPase Rac1 at sites where fine filopodia mediate the initial contact between myoblasts. In addition, DGK-zeta codistributed with the Ca(2+)-dependent cell adhesion molecule N cadherin at nascent, but not previously established cell contacts. We provide evidence that C2 cells are pulled together at cell-cell junctions by N-cadherin containing filopodia reminiscent of epithelial adhesion zippers, which guide the advance of lamellipodia from apposing cells. At later times, vesicles with properties of macropinosomes formed close to cell-cell junctions. Reconstruction of confocal optical sections showed these form dome-like protrusions from the dorsal surface of contacting cells. Collectively, these results suggest DGK-zeta and syntrophins play a role at multiple stages of the fusion process. Moreover, our findings provide a potential link between changes in the lipid content of the membrane bilayer and reorganization of the actin cytoskeleton during myoblast fusion. PMID- 17410542 TI - The Drosophila nucleoporin gene nup154 is required for correct microfilament dynamics and cell death during oogenesis. AB - The Drosophila nucleoporin gene nup154 is required in both male and female germline for successful gametogenesis. Mutant flies lack differentiated sperm and lay abnormal eggs. We demonstrated that the egg phenotype was associated with specific alterations of the actin cytoskeleton at different stages of oogenesis. Actually, mutant egg chambers displayed an abnormal organization of both subcortical microfilaments and cytoplasmic actin bundles, that led to defective nurse cell dumping. TUNEL analysis also showed that the dumpless phenotype was associated with delayed apoptosis. The nup154 gene product was localized by conventional immunofluorescence microscopy to the nuclear envelope in a distinct punctuate pattern, characteristic of nuclear pore complex components. TEM analysis revealed that the protein was mainly distributed along filamentous structures that extended radially on the nuclear side of the pore, suggesting that Nup154 could be an integral component of the basket filaments associated with the nuclear pore complexes. We propose that Nup154 is necessary for correct nuclear pore complex functions and that the proper regulation of the actin cytoskeleton dynamics strongly relies upon nuclear pore integrity. PMID- 17410544 TI - Nucleolar development and allocation of key nucleolar proteins require de novo transcription in bovine embryos. AB - The goal of the present study was to investigate whether key nucleolar proteins involved in ribosomal RNA (rRNA) transcription and processing are transcribed de novo or from maternally inherited messenger RNAs (mRNA) in bovine embryos, and to which extent de novo transcription of these proteins mRNA is required for the development of functional nucleoli during the major activation of the embryonic genome. Immunofluorescence for localization of key nucleolar proteins, autoradiography for detection of transcriptional activity, and transmission electron microscopy were applied to in vitro produced bovine embryos cultured from the 2-cell stage with or without (control groups) alpha-amanitin, which blocks the RNA polymerases II and III transcription and, thus the synthesis of mRNA. In the control groups, weak autoradiographic labeling was initially observed in the periphery of few nuclei at the 4-cell and the early 8-cell stage, and the entire nucleoplasm as well as nucleolus precursor bodies (NBBs) were prominently labelled in all late 8-cell stages. The NPBs displayed initial transformation into fibrillo-granular nucleoli. In the alpha-amanitin group, lack of autoradiographic labeling was seen at all developmental stages and disintegrated NPBs stage were found at the late 8-cell. Our immunofluorescence data indicate that RNA polymerase I, UBF, topoisomerase I and fibrillarin are transcribed de novo whereas nucleolin and nucleophosmin are maternally inherited as demonstrated by alpha -amanitin inhibition. However, localization of these two proteins to the nucleolar compartments was negatively affected by the alpha amanitin treatment. Consequently, functional nucleoli were not established. PMID- 17410545 TI - Presumptive pre-Sertoli cells express genes involved in cell proliferation and cell signalling during a critical window in early testis differentiation. AB - In mammals, the pre-Sertoli cell of the male genital ridge is the first cell type to display sex specific differentiation and differential gene expression. The genetic cascade driving the differentiation of pre-Sertoli cells and ultimately testis formation is beginning to be unravelled, but many questions remain. A better understanding of the transcriptome of pre-Sertoli cells immediately after sex determination is essential in order to further understand this differentiation process. A mouse model expressing Red Fluorescent Protein (RFP) under the control of a hybrid mouse/pig SRY promoter (HybSRYp-RFP) was used to purify cells from embryonic day 12.0 (e12.0) male genital ridges. To compare the transcriptomes of HybSRYp-RFP cell populations versus age matched whole female genital ridges, RNA was extracted and used to generate molecular probes that were hybridized onto Affymetrix Mouse Genome 430 2.0 micro-arrays. The expression of genes considered markers for pre-Sertoli cells, including Sox9, Mis, Dhh and Fgf9 were identified within the HybSRYp-RFP expressing cell population, while markers for germ cells (Oct4, SSEA-1) and endothelial cells (Ntrk3) were not identified. In contrast, markers for ovarian somatic cell expression, including Fst and Bmp2, were identified as overexpressed within the ovarian cell population. In a general fashion, genes identified as 2.5-fold over expressed in HybSRYp-RFP expressing cells coded notably for cell signalling and extra cellular proteins. The expression of Sox10, Stc2, Fgf18, Fgf13 and Wnt6 were further characterized via whole mount in situ hybridization (WISH) on male and female genital ridges between e11.5 and e14.5. Sox10, Fgf18, Fgf13 and Stc2 gene expression was detected within the male genital ridges while Wnt6 was found diffusely within both the male and female genital ridges. These data represent the earliest comprehensive microarray expression analysis of purified presumptive pre-Sertoli cells available to date. PMID- 17410546 TI - Increase of the trophoblast giant cells with prolactin-releasing peptide (PrRP) receptor expression in p53-null mice. AB - Trophoblast giant cells in the mouse placentas are polyploid cells that form as a result of endoreduplication. The giant cells form the outermost layer of the extraembryonic compartment and produce a number of pregnancy-specific hormones, including prolactin family members. Here we demonstrate that trophoblast giant cells are increased, and display upregulation of prolactin releasing peptide (PrRP) receptor in the p53-null (p53(-/-)) embryonic placentas. At day 13.5 of gestation, the weight of p53(-/-) placentas was less than that of both wild-type and p53(+/-) placentas. In p53(-/-) placentas, the spongiotrophoblast layer was significantly decreased in thickness, and the trophoblast giant cells were observed not only in the outer layer of placentas but in both the spongiotrophoblast layer and the labyrinthine layer. The giant cells spread over the spongiotrophoblast and labyrinthine layer in p53(-/-) placentas displayed more intensive expression of immunoreactive PrRP receptor than in wild-type placentas. Previous studies indicated that the association between PrRP and PrRP receptor physiologically involves in the expression and secretion of the peptide hormones, including prolactin and growth hormones. These results suggest that p53 may regulate the differentiation of trophoblast giant cells, and may control the physiological PrRP stimuli in mouse placentas. PMID- 17410547 TI - Effects of calcitonin and parathyroid hormone on the regulation of cabindin-D(9k) in the uterus, placenta, and fetal membrane of rats related to blood calcium level during late gestation. AB - Calbindin-D(9k) (CaBP-9k) gene is expressed in the uterus of pregnant rats, which is regulated by steroid hormones during estrous cycle or gestation. We hypothesized that there is a positive correlation between altered CaBP-9k expression and change in one or more of the hormones to provide a clue to the mechanism responsible for the altered calcium levels in the uterus, placenta, and fetal membrane during late gestation. Thus, in the present study, we investigated the effects of the hormones including estradiol (E2), calcitonin (CT), and parathyroid hormone (PTH) on the regulation of CaBP-9k in these tissues. There was an increase in the level of CaBP-9k in the uterus, placenta, and extra embryonic membrane at late gestation, as blood calcium level increased. The protein level of CaBP-9k remained lower in the uterus at two-thirds of pregnancy, and then it rebounded abruptly during late pregnancy. During late gestation, E2 is postulated to be a dominant factor in the regulation of uterine CaBP-9k gene expression. Furthermore, we assumed that there is a positive correlation between altered expression of CaBP-9k and blood calcium level during pregnancy. The present study demonstrated the regulation of CaBP-9k mRNA in the uterus, placenta, and fetal membrane of rats, implying a role for CaBP-9k gene in the control of blood calcium in placenta and the calcium passing from maternal blood to fetal circulation. Taken together, these results suggest that major alterations in calcium metabolism caused by maternal thyroparathyroidectomy (TPTX), are sufficient to affect the changes in reproductive tissues during late pregnancy. In addition, an increase of blood calcium level is one of the most significant factors in the regulation of CaBP-9k at the transcriptional and/or translational levels in the reproductive tissues during late pregnancy. PMID- 17410548 TI - Abnormal lens morphogenesis and ectopic lens formation in the absence of beta catenin function. AB - beta-Catenin plays a key role in cadherin-mediated cell adhesion as well as in canonical Wnt signaling. To study the role of beta-catenin during eye development, we used conditional Cre/loxP system in mouse to inactivate beta catenin in developing lens and retina. Inactivation of beta-catenin does not suppress lens fate, but instead results in abnormal morphogenesis of the lens. Using BAT-gal reporter mice, we show that beta-catenin-mediated Wnt signaling is notably absent from lens and neuroretina throughout eye development. The observed defect is therefore likely due to the cytoskeletal role of beta-catenin, and is accompanied by impaired epithelial cell adhesion. In contrast, inactivation of beta-catenin in the nasal ectoderm, an area with active Wnt signaling, results in formation of crystallin-positive ectopic lentoid bodies. These data suggest that, outside of the normal lens, beta-catenin functions as a coactivator of canonical Wnt signaling to suppress lens fate. PMID- 17410549 TI - Tool use during display behavior in wild Cross River gorillas. AB - During the course of a 3-year ecological study on Cross River gorillas (Gorilla gorilla diehli) at the Kagwene Mountain in Cameroon, we observed three cases of tool use which may be unique to the gorillas of this region and possibly learned through interactions with humans. A non-habituated group of Cross River gorillas threw fistfuls of grass toward humans in display contexts. An individual gorilla was also observed to throw a detached branch toward researchers during another encounter. The third encounter occurred between the focal group and a local man, in which the man threw stones at the gorillas and they reacted by throwing fistfuls of grass back at him. We describe these observations and speculate that this sub-population of gorillas developed this display behavior as a response to particular local conditions. PMID- 17410550 TI - Characterization of the RNA binding energetics of the Candida albicans poly(A) polymerase. AB - The 3' ends of eukaryotic mRNAs are characterized by the presence of a poly(A) tail, which plays a critical role in stability, transport, and translation of the mRNAs. In the present study, we report the expression, purification and enzymatic characterization of the poly(A) polymerase of Candida albicans, an important human pathogen. As a first step toward elucidating the nature of the interaction between RNA and the enzyme, fluorescence spectroscopy assays were also performed to monitor the binding of RNA to the protein. Our assays revealed that the initial interaction between RNA and the enzyme is characterized by a high enthalpy of association and that the minimal RNA binding site of the enzyme is eight nucleotides. Moreover, both the kinetics of real-time RNA binding and the contribution of electrostatic interactions to the overall binding energy were investigated. Finally, we also correlated the effect of RNA binding on protein structure, using both circular dichroism and guanidium hydrochloride-induced denaturation studies as structural indicators. Our data indicate that the protein undergoes structural modifications upon RNA binding, although the interaction does not significantly modify the stability of the protein. In addition to the determination of the energetics of RNA binding, our study provides a better understanding of the molecular basis of RNA binding by poly(A) polymerases. PMID- 17410551 TI - Determination of ruthenium originating from the investigational anti-cancer drug NAMI-A in human plasma ultrafiltrate, plasma, and urine by inductively coupled plasma mass spectrometry. AB - We present a highly sensitive, rapid method for the determination of ruthenium originating from the investigational anti-cancer drug NAMI-A in human plasma ultrafiltrate, plasma, and urine. The method is based on the quantification of ruthenium by inductively coupled plasma mass spectrometry and allows quantification of 30 ng L(-1) ruthenium in plasma ultrafiltrate and urine, and 75 ng L(-1) ruthenium in human plasma, in 150 microL of matrix. The sample pretreatment procedure is straightforward and only involves dilution with appropriate diluents. The performance of the method, in terms of accuracy and precision, fulfilled the most recent FDA guidelines for bioanalytical method validation. Validated ranges of quantification were 30.0 to 1 x 10(4) ng L(-1) for ruthenium in plasma ultrafiltrate and urine and 75.0 to 1 x 10(4) ng L(-1) for ruthenium in plasma. The applicability of the method and its superiority to atomic-absorption spectrometry were demonstrated in two patients who were treated with intravenous NAMI-A in a phase I trial. The assay is now successfully used to support pharmacokinetic studies in cancer patients treated with NAMI-A. PMID- 17410552 TI - Stable strontium isotopic ratios from archaeological organic remains from the Thorsberg peat bog. AB - Stable strontium isotope ratios in archaeological finds have frequently been used to determine their place of origin, in order to reconstruct migration and trade. Peat bogs offer favourable burial conditions for the preservation of organic remains such as woollen textiles and leather by a natural tanning process. However, these finds are impregnated by peat substances including contaminant strontium which is likely to mask the original (87)Sr/(86)Sr isotopic ratio of the specimens. In this paper, we present a pilot study analysing stable strontium isotopic ratios from Iron Age textile and leather finds from the Thorsberg peat bog, focusing on a sample processing method which permits the quantitative removal of contaminating strontium from the specimens. PMID- 17410553 TI - Optimal selection of markers for validation or replication from genome-wide association studies. AB - With reductions in genotyping costs and the fast pace of improvements in genotyping technology, it is not uncommon for the individuals in a single study to undergo genotyping using several different platforms, where each platform may contain different numbers of markers selected via different criteria. For example, a set of cases and controls may be genotyped at markers in a small set of carefully selected candidate genes, and shortly thereafter, the same cases and controls may be used for a genome-wide single nucleotide polymorphism (SNP) association study. After such initial investigations, often, a subset of "interesting" markers is selected for validation or replication. Specifically, by validation, we refer to the investigation of associations between the selected subset of markers and the disease in independent data. However, it is not obvious how to choose the best set of markers for this validation. There may be a prior expectation that some sets of genotyping data are more likely to contain real associations. For example, it may be more likely for markers in plausible candidate genes to show disease associations than markers in a genome-wide scan. Hence, it would be desirable to select proportionally more markers from the candidate gene set. When a fixed number of markers are selected for validation, we propose an approach for identifying an optimal marker-selection configuration by basing the approach on minimizing the stratified false discovery rate. We illustrate this approach using a case-control study of colorectal cancer from Ontario, Canada, and we show that this approach leads to substantial reductions in the estimated false discovery rates in the Ontario dataset for the selected markers, as well as reductions in the expected false discovery rates for the proposed validation dataset. PMID- 17410554 TI - Testing association between disease and multiple SNPs in a candidate gene. AB - Current technology allows investigators to obtain genotypes at multiple single nucleotide polymorphism (SNPs) within a candidate locus. Many approaches have been developed for using such data in a test of association with disease, ranging from genotype-based to haplotype-based tests. We develop a new approach that involves two basic steps. In the first step, we use principal components (PCs) analysis to compute combinations of SNPs that capture the underlying correlation structure within the locus. The second step uses the PCs directly in a test of disease association. The PC approach captures linkage-disequilibrium information within a candidate region, but does not require the difficult computing implicit in a haplotype analysis. We demonstrate by simulation that the PC approach is typically as or more powerful than both genotype- and haplotype-based approaches. We also analyze association between respiratory symptoms in children and four SNPs in the Glutathione-S-Transferase P1 locus, based on data from the Children's Health Study. We observe stronger evidence of an association using the PC approach (p = 0.044) than using either a genotype-based (p = 0.13) or haplotype based (p = 0.052) approach. PMID- 17410555 TI - Applications of mass spectrometry for the structural characterization of recombinant protein pharmaceuticals. AB - Therapeutic proteins produced using recombinant DNA technologies are generally complex, heterogeneous, and subject to a variety of enzymatic or chemical modifications during expression, purification, and long-term storage. The use of mass spectrometry (MS) for the evaluation of recombinant protein sequence and structure provides detailed information regarding amino acid modifications and sequence alterations that have the potential to affect the safety and activity of therapeutic protein products. General MS approaches for the characterization of recombinant therapeutic protein products will be reviewed with particular attention given to the standard MS tools available in most biotechnology laboratories. A number of recent examples will be used to illustrate the utility of MS strategies for evaluation of recombinant protein heterogeneity resulting from post-translational modifications (PTMs), sequence variations generated from proteolysis or transcriptional/translational errors, and degradation products which are formed during processing or final product storage. Specific attention will be given to the MS characterization of monoclonal antibodies as a model system for large, glycosylated, recombinant proteins. Detailed examples highlighting the use of MS for the analysis of monoclonal antibody glycosylation, deamidation, and disulfide mapping will be used to illustrate the application of these techniques to a wide variety of heterogeneous therapeutic protein products. The potential use of MS to support the selection of cell line/clone selection and formulation development for therapeutic antibody products will also be discussed. PMID- 17410557 TI - Strip-track revascularization after stripping of the great saphenous vein. AB - BACKGROUND: Varicose veins that recur after standard high tie and strip are often associated with venous reflux in the thigh, as shown by duplex ultrasonography. The aim of this study was to look for evidence of revascularization in the strip track after great saphenous vein (GSV) stripping. METHODS: A consecutive series of patients with duplex-proven great saphenous varicose veins underwent saphenofemoral ligation with intraoperative confirmation of successful stripping. Duplex ultrasonography was performed 1 week and 1 year after surgery. The presence and extent of haematoma was noted, as was any venous reflux within the strip track. RESULTS: At 1 year, four (6 per cent) of 70 patients had complete revascularization of the strip track and 12 (17 per cent) had partial revascularization, all with duplex-proven reflux. Partial revascularization was in the distal third of the track in six legs (9 per cent of the 70 patients), in the distal half in five (7 per cent) and was almost complete in one leg (1 per cent). All patients with revascularization had a significant strip-track haematoma at 1 week after surgery. CONCLUSION: Revascularization of the GSV strip track after stripping was found in 23 per cent of patients after 1 year; all of these had a postoperative haematoma in the track. PMID- 17410558 TI - Sacral nerve stimulation for neurogenic faecal incontinence. AB - BACKGROUND: Sacral nerve stimulation (SNS) has emerged as a promising technique for the treatment of faecal incontinence. This study assessed the outcome of SNS in a cohort of patients with incontinence of neurological aetiology. METHODS: Thirty-six patients were included in a trial of SNS. Twenty-nine subsequently had a permanent implant. Evaluation consisted of a continence diary, anal manometry, saline retention testing and quality of life assessment. RESULTS: After a median follow-up of 35 (range 3-71) months, 28 patients showed a marked improvement in or complete recovery of continence. Incontinence to solid or liquid stool decreased from a median of 7 (range 4-15) to 2 (range 0-5) episodes in 21 days (P = 0.002). Saline retention time increased from a median of 2 (range 0-5) to 7 (range 2-15) min (P = 0.002). Maximum resting and squeeze anal canal pressures increased compared with preoperative values. Quality of life on all scales among patients who received a permanent implant increased at 12 and 24 months after operation. CONCLUSION: SNS is of value in selected patients with neurogenic faecal incontinence. PMID- 17410559 TI - Desmoid tumours in patients with familial adenomatous polyposis and desmoid region adenomatous polyposis coli mutations. AB - BACKGROUND: :The aim of this study was to determine the proportion of patients with familial adenomatous polyposis (FAP) who had mutations in the desmoid region of the adenomatous polyposis coli (APC) gene that phenotypically expresses desmoid disease, and to determine the role for surgery in these patients. METHODS: Data from the North West Region FAP database and case notes were analysed retrospectively. RESULTS: Of 363 patients with FAP, 47 from ten families had APC mutations in the desmoid region 3' to codon 1399. Of 22 patients undergoing surgery, 16 developed desmoids, and of these 12 had mesenteric desmoid disease. Complications from mesenteric desmoids were death (two patients), enterectomy (three), local resection (three), fistula (one), cholangitis and local resection (one), bowel obstruction (one) and bowel and ureteric obstruction (one). Preoperative polyp burden ranged from 0 to 100 in eight patients (median age 24.5 (range 16-39) years) and more than 100 in seven (median age 39 (range 31 64) years). One patient had no record of polyp burden. CONCLUSION: In individuals with 3' APC mutations, abdominal surgery is associated with a 65 per cent risk of developing mesenteric desmoids. An alternative strategy might be to attempt to manage the polyps endoscopically. PMID- 17410560 TI - Quadrature coil design for high-resolution carotid artery imaging scores better than a dual phased-array coil design with the same volume coverage. AB - PURPOSE: To evaluate the ability of a custom-built coil design to provide improved signal-to-noise ratio (SNR) and less signal drop with increasing depth at the carotid artery. MATERIALS AND METHODS: Phased-array surface coils can provide a high SNR to image the carotid vessel wall. However, given the required field-of-view (FOV) and penetration depth, these coils show either a fast signal drop with increasing depth or a moderate SNR at increased coil size. A quadrature surface coil (a butterfly coil in conjunction with a linear single-loop coil) was compared with a phased-array coil in phantom and human studies. RESULTS: The phantom studies showed that the quadrature coil has better SNR over the required FOV than a standard phased-array coil (26% at 3 cm depth). CONCLUSION: The quadrature coil enables better image quality to be achieved. PMID- 17410561 TI - Functional lung imaging using hyperpolarized gas MRI. AB - The noninvasive assessment of lung function using imaging is increasingly of interest for the study of lung diseases, including chronic obstructive pulmonary disease (COPD) and asthma. Hyperpolarized gas MRI (HP MRI) has demonstrated the ability to detect changes in ventilation, perfusion, and lung microstructure that appear to be associated with both normal lung development and disease progression. The physical characteristics of HP gases and their application to MRI are presented with an emphasis on current applications. Clinical investigations using HP MRI to study asthma, COPD, cystic fibrosis, pediatric chronic lung disease, and lung transplant are reviewed. Recent advances in polarization, pulse sequence development for imaging with Xe-129, and prototype low magnetic field systems dedicated to lung imaging are highlighted as areas of future development for this rapidly evolving technology. PMID- 17410562 TI - MRCP imaging at 3.0 T vs. 1.5 T: preliminary experience in healthy volunteers. AB - PURPOSE: To evaluate the impact of magnetic resonance cholangiopancreatography (MRCP) imaging at 1.5T and 3.0T on image quality. MATERIALS AND METHODS: Fourteen volunteers were examined at both 1.5T and 3.0T using MRCP imaging performed with a breath-held two-dimensional (2D) half-Fourier acquired single-shot turbo spin echo (HASTE) thick-slab sequence, a free-breathing navigator-triggered three dimensional (3D) turbo spin-echo (TSE) sequence with prospective acquisition correction, and a heavily T2-weighted (T2W) sequence with breath-held multislice HASTE. All images were scored for visualization of the biliary and pancreatic ducts, severity of artifacts, image noise, and overall image quality. RESULTS: MRCP imaging at 3.0T yielded a significant improvement in overall image quality compared to 1.5T. We found a trend for superior visualization of the biliary and pancreatic ducts at 3.0T. Heavily T2W imaging with thin sections (1.4 mm) at 3.0T provided diagnostic images and better visualization of the biliary and pancreatic ducts than heavily T2W imaging with standard sections (2.8 mm) at 3.0T. CONCLUSION: Our experience suggests that MRCP imaging at 3.0T has the potential to provide excellent images. High-resolution heavily T2W imaging with a small voxel size (1.3 x 1.3 x 1.4 mm) at 3.0T can provide diagnostic images and allow evaluation of small pathologies of the bile and pancreatic ducts, which 1.5T MRI cannot sufficiently visualize. PMID- 17410563 TI - Transient signal changes in diffusion-weighted stimulated echoes during neuronal stimulation at 3T. AB - PURPOSE: To develop a sensitive method for detecting minute transient signal changes that can arise due to variations in the extravascular apparent self diffusion coefficient, D, during neuronal activation. MATERIALS AND METHODS: A three-pulse sequence that reads out a moderately diffusion-weighted (DW) primary echo (PRE) and a heavily DW stimulated echo (STE) was employed to investigate whether small transient signal changes in extravascular D occur in response to a visual stimulus. Contributions to signal changes caused by subtle differences in the transient variations of the apparent transverse relaxation constant, T(2), between the PRE and STE were also quantified. RESULTS: On z-maps obtained from the STE, more voxels showed significant stimulus-related signal changes compared to maps of the PRE. The average maximum signal change of the STE was larger than that of the PRE. The observed increase in the relative signal change was independent of the strength of the diffusion weighting. CONCLUSION: The STE is more sensitive to neuronal activity than the PRE. The discrepancy between the two echoes does not arise from transient changes in D, but from subtle differences in stimulus-related variations of T(2) between the two echoes. PMID- 17410564 TI - Safety and effectiveness of long-term budesonide treatment in maintaining remission in patients with mild-to-moderate Crohn's disease. PMID- 17410565 TI - Time-resolved contrast-enhanced carotid imaging using undersampled projection reconstruction acquisition. AB - PURPOSE: To investigate the utility of nonuniform angular spacing of projections in a three-dimensional (3D) hybrid undersampled projection reconstruction (PR) acquisition for contrast-enhanced (CE) time-resolved carotid imaging. MATERIALS AND METHODS: Carotid CE magnetic resonance angiography (CE-MRA) was performed on seven healthy volunteers using a time-resolved hybrid sequence that combined undersampled PR acquisition in-plane and Cartesian slice encoding through-plane. The undersampling streak artifact comes mainly from the superior-inferior (S/I) direction in carotid imaging and is suppressed by nonuniform distribution of the projections. Phantom and volunteer studies were performed to demonstrate its efficacy. RESULTS: The undersampling streak artifact was significantly suppressed through a nonuniform distribution of the projection angles with more projections aligned along the S/I direction. The hybrid PR sequence combined with nonuniform distribution of the projection angles provided time-resolved images of the carotid arteries with high temporal resolution (two seconds per frame) and high spatial resolution (1.0 x 1.0 x 1.5 mm(3)) simultaneously. CONCLUSION: High resolution dynamic imaging of the carotid arteries is feasible with the use of a hybrid undersampled PR acquisition. Undersampling streak artifact can be suppressed significantly through nonuniform distribution of the projections. PMID- 17410566 TI - Calf muscle perfusion at peak exercise in peripheral arterial disease: measurement by first-pass contrast-enhanced magnetic resonance imaging. AB - PURPOSE: To develop a contrast-enhanced magnetic resonance (MR) technique to measure skeletal muscle perfusion in peripheral arterial disease (PAD). MATERIALS AND METHODS: A total of 11 patients (age = 61 +/- 11 years) with mild to moderate symptomatic PAD (ankle-brachial index [ABI] = 0.75 +/- 0.08) and 22 normals were studied using an MR-compatible ergometer. PAD and normal(max) (Nl(max); N = 11) exercised to exhaustion. Nl(low) (N = 11) exercised to the same workload achieved by PAD. At peak exercise, 0.1 mm/kg of gadolinium diethylenetriamine pentaacetic acid (Gd-DTPA) was infused at 3-4 cm(3)/second followed by a saline flush at the same rate. A dual-contrast gradient echo (GRE) sequence enabled simultaneous acquisition of muscle perfusion and arterial input function (AIF). The perfusion index (PI) was defined as the slope of the time-intensity curve (TIC) in muscle divided by the arterial TIC slope. RESULTS: Median workload was 120 Joules in PAD, 210 Joules in Nl(low), and 698 Joules in Nl(max) (P < 0.001 vs. Nl(low) and PAD). Median PI was 0.29 in PAD (25th and 75th percentiles [%] = 0.20, 0.40), 0.48 in Nl(low) (25th, 75th % = 0.36, 0.62; P < 0.02 vs. PAD), and 0.69 in Nl(max) (25th, 75th % = 0.5, 0.77; P < 0.001 vs. PAD). Area under the ROC-curve for PI differentiating patients from Nl(max) was 0.95 (95% confidence interval [CI] = 0.77-0.99). CONCLUSION: Peak-exercise measurement of lower limb perfusion with dual-contrast, first-pass MR distinguishes PAD from normals. This method may be useful in the study of novel therapies for PAD. PMID- 17410567 TI - Comparative assessment of hardening of demineralized dentin under lining materials using an ultramicroindentation system. AB - The aim of the current in vitro study was to evaluate the influence of three lining materials with a reported mineralizing capacity on hardness and elasticity of demineralized dentin. Four standardized microcavities were prepared in exposed dentin surfaces of 16 extracted human molars each. Dentin was demineralized in 0.5M EDTA for 2 h. One microcavity was left empty. The others were filled with a resin-modified glass ionomer cement (RMGIC), a bioactive glass S53P4 suspension, and a prototype Ca-PO(4) cement. Teeth were then immersed in deionized water or simulated oral fluid. After 3 weeks, hardness and composite elastic modulus of the dentin subjacent to the microcavities were assessed under wet conditions using the ultramicroindentation system (UMIS). After immersion in deionized water, there was no significant improvement of the mechanical properties of dentin irrespective of the material applied beforehand, indicating a lack of direct material effects. Exposure to simulated oral fluid resulted in significantly (p < 0.05) higher hardness and composite elastic modulus values of the dentin subjacent to empty microcavities and counterparts lined with bioactive glass compared to corresponding dentin under the RMGIC. UMIS profiles showed little variance. PMID- 17410568 TI - Development and in vivo testing of a Nitinol tracheal stent. AB - This article describes the development of a Nitinol tracheal stent (HCPA NiTi stent) and its application in a feline animal model. Straight-annealed, bright polished Nitinol wire (55.8%Ni-44.2%Ti) was weaved around a 40-mm-long metal fixture with 8-mm diameter. The prototypes were submitted to different times of shape-setting heat treatment (530 degrees C), which resulted in stents of different colors and caused some variation in length and diameter. The prototypes were then submitted to compression testing, and the most resistant pieces, requiring the greatest force to achieve a 25% reduction in diameter and presenting the least variation in length and diameter (dark blue, 9 min of heat treatment), were submitted to fatigue testing. After that, only dark blue stents were manufactured and implanted in felines. No migration, tracheal stenosis, or any other type of damage were observed after 40 weeks. The integrity of the tracheal wall in contact with the stent was confirmed by macro and microscopic analyses. The development and in vivo testing of the HCPA NiTi-stent were successful. PMID- 17410569 TI - Development of the wing-attached rod for acceleration of "Biotube" vascular grafts fabrication in vivo. AB - To accelerate the fabrication of in vivo tissue-engineered autologous vascular prosthetic tissues, the "Biotube," a novel wing-attached rod mold was designed for a tissue rolling technique based on a two-step in body tissue incubation (IBTI) process. The new mold consisted of a silicone rod (3-mm diameter, 23-mm length) partly connected to a poly(ethylene terephthalate) film (a wing, 23 x 19 x 0.1 mm). While the molds were embedded into the dorsal subcutaneous pouches of rabbits for 2 weeks (primary IBTI), they were encapsulated fully with thin connective tissues. After removal of the wing materials, the remaining saccular membranous tissues were rolled up on the core tubular tissues that had formed around the silicone rods. Following another 2-week embedding of the assembled tissues (secondary IBTI), the layered tissues fused to each other to form compliant and stiff tubular tissues, "Rolled Biotubes." The wall thickness of the Rolled Biotubes was about 800 microm and the burst strength was about 4000 mmHg, both of which were significantly higher than those of Biotubes prepared by one step, 4-week IBTI or two-step, 2-week IBTI (p < 0.05). A Rolled Biotube could be applied as middle or large caliber arterial prostheses. PMID- 17410570 TI - Adhesive performance of dentin bonding agents applied in vivo and in vitro. Effect of intrapulpal pressure and dentin depth. AB - The purpose of this study was to evaluate the influence of intrapulpal pressure and dentin depth on bond strengths of an etch-and-rinse and a self-etching bonding agent to dentin in vitro and in vivo. Twenty-four pairs of premolars were randomly divided into four groups (n = 6) according to the dentin bonding agent, Single Bond and Clearfil SE Bond, and intrapulpal pressure, null or positive. Each tooth of the pair was further designated to be treated in vivo or in vitro. The intrapulpal pressure was controlled in vivo by the delivery of local anesthetics containing or not a vasoconstrictor, while in vitro, it was achieved by keeping the teeth under hydrostatic pressure. Class I cavities were prepared and the dentin bonding agents were applied followed by incremental resin restoration. For the teeth treated in vitro, the same restorative procedures were performed after a 6 month-storage period. Beams with 1 mm(2) cross-sectional area were prepared and microtensile tested. Clearfil SE Bond was not influenced by any of the variables of the study, while bond strengths produced in vitro were significantly higher for Single Bond. Overall, lower bond strengths were produced in deep dentin, which reached statistical significance when Single Bond was applied under physiological or simulated intrapulpal pressure. In conclusion, in vitro bonding may overestimate the immediate adhesive performance of more technique-sensitive dentin bonding systems. The impact of intrapulpal pressure on bond strength seems to be more adhesive dependent than dentin morphological characteristics related to depth. PMID- 17410571 TI - Chitosan/poly(vinyl alcohol) blending hydrogel coating improves the surface characteristics of segmented polyurethane urethral catheters. AB - Segmented polyurethane (SPU) is commonly used to manufacture urethral catheters. Surface modifications for SPU catheters are needed to reduce friction and protein adsorption, in order to minimize catheter-related complications, including urethral trauma, encrustation, catheter obstruction, bacterial colonization, and infection. In this study, a four-step surface modification method was developed to create a thin lubricious layer of chitosan/poly(vinyl alcohol) (PVA) hydrogel on the SPU catheter. Modification steps included oxidation of the SPU surface, functionalities modification, carbodiimide reaction and coupling, and hydrogel crosslinking. The success of each modification step was confirmed by Fourier transform infrared spectroscopy. Measurement of the water contact angle revealed that hydrogel coating created a highly hydrophilic surface and atomic force microscope analyses demonstrated that the surface was slippery. Protein absorption of the SPU catheter was significantly reduced by coating hydrogel. Chitosan in the hydrogel could provide antimicrobial activity, and the hydrogel coating SPU samples showed significant antibacterial effects in this study. In summary, the four-step modification method developed in this study provided a simple and effective way to coat the surface of SPU catheters with a chitosan/PVA blending hydrogel that could help to minimize the risk of complications related to the use of urethral catheters. PMID- 17410572 TI - Effects of a ferric chloride primer on collagen-depleted dentin bonding between tri-n-butylborane initiated self-curing resin and dentin. AB - This study was designed to evaluate the micro-tensile bond strength between a carboxylic resin and dentin, when the dentin surface was modified with an experimental dentin primer. The three primers tested were ED primer II (ED), 0.3% ferric chloride aqueous solution (FE), and ED containing 0.3% ferric chloride (ED/FE). Three commercial dentin conditioners [40% phosphoric acid, 10% NaOCl, and 10% citric acid with 3% ferric chloride (10-3)] were also used. The coronal surfaces of extracted human molars were ground flat to dentin. The dentin surfaces were treated with phosphoric acid, NaOCl, or with one of the primers. The 10-3 was used without phosphoric acid or NaOCl as a control. A composite material rod was bonded to the dentin surface with 4-META/MMA-TBB resin. After 24 h immersion in 37 degrees C distilled water, 0.9 mm x 0.9 mm composite-dentin beams cut from the bonded specimens were stressed to failure in tension at 0.6 mm/min. The bond strengths were also evaluated after 5000 thermocycles. The bond strength of the group ED/FE was significantly higher than those of the 10-3, ED, and FE. After 5000 thermocycling, 10-3, ED and FE showed significant decrease in bond strength, although no significant decrease was seen for ED/FE. It was concluded that dentin surface treatment with phosphoric acid, NaOCl, and the ED/FE primer improved the bonding (p < 0.05) between 4-META/MMA-TBB resin and dentin, with or without thermocycling, while the bond strengths in the control group fell 34% following 5000 thermocycles. PMID- 17410573 TI - Comparison of amorphous TCP nanoparticles to micron-sized alpha-TCP as starting materials for calcium phosphate cements. AB - The development of degradable bone cements with a mineral composition similar to natural bone was investigated using highly reactive calcium phosphate phases as starting materials. Mixtures of XRD-amorphous, glassy tricalcium phosphate (amorphous-TCP) nanoparticles of 25-60 nm size and micron sized, milled alpha-TCP were set by hydration with sodium phosphate buffer and investigated for possible application as single component calcium phosphate cements (CPCs). Isothermal calorimetry allowed a precise tracking of the setting process. Amorphous-TCP nanoparticles converted into calcium deficient hydroxyapatite with cement setting times below 12 min. The total energy release by the material during hardening corroborated the importance of high specific surface area and phase composition, that is, amorphous state of the nanometric starting material as repeatedly suggested earlier. The phase composition of the resulting CPCs was characterized by X-ray diffraction before and after setting. The morphology was investigated by nitrogen adsorption, scanning, and transmission electron microscopy and revealed the formation of highly porous calcium deficient hydroxyapatite with specific surface areas of up to 160 m(2) g(-1) after setting. In contrast to the very fast reaction time and highest specific surface area, the mechanical stability of the resulting CPC is still insufficient and requires further improvement. PMID- 17410574 TI - Ofloxacin-delivery system of a polyanhydride and polylactide blend used in the treatment of bone infection. AB - We developed a local drug-release system consisting of two biodegradable polymers, poly(sebacic anhydride) (PSA) and poly-D,L-lactide (PLA), for the treatment of chronic osteomyelitis. PSA and PLA were dissolved and blended at different ratios in tetrahydrofuran. Ofloxacin was loaded with an 8:1 weight ratio of the blend to the drug. The ofloxacin-containing beads of the PSA/PLA blend were made by preheating and compressing them in a mold. The in vitro drug release showed that changing the ratio between the two polymers caused the effective ofloxacin-release duration to vary from 6 to 68 days. The ofloxacin containing beads with 10% PSA and 90% PLA produced an inhibition zone for the bacteria Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa within 89 days of the experiment. The in vivo drug release of the beads in rabbits demonstrated that the average ofloxacin concentration in the local bone was 20.1 +/- 10.3 microg/g, while that in the plasma was 35.6 +/- 18.8 ng/mL, within 8 weeks. Roentgenography, bacterial cultures, and histological examinations showed that the local release of ofloxacin by the beads could cure osteomyelitis in rabbits. Our findings suggested that using PSA/PLA blends with different ratios as carriers for antibiotics might be useful in the treatment of chronic osteomyelitis and in the prophylaxis of bone infection. PMID- 17410575 TI - Dynamic MR assessment of the anorectal angle and puborectalis muscle in pediatric patients with anismus: technique and feasibility. AB - PURPOSE: To assess the feasibility of dynamic breath-hold MRI for evaluating changes in the anorectal angle and movements of the pelvic-floor musculature (puborectalis) during resting and straining states in pediatric patients presenting with anismus. MATERIALS AND METHODS: Six pediatric patients (7-13 years old) with chronic constipation and manometric evidence of anismus were assessed by dynamic breath-hold MRI. Changes in the anorectal angle, the degree of pelvic-floor descent, and the thickness and length of the puborectalis muscles were measured during rest and straining. The findings were compared with those obtained in six age- and sex-matched controls. RESULTS: The children with anismus had a smaller anorectal angle during straining, and the angle decreased from rest to defecation. The puborectalis also became paradoxically shortened and thickened during straining in the anismus group. There were significant differences between the two groups in terms of the change of degree of the anorectal angle, and the thickness and length of the puborectalis muscle during straining. CONCLUSION: Fast dynamic MRI is feasible for evaluating pelvic-floor movement in pediatric patients. Preliminary results suggest that children with anismus have a smaller anorectal angle and a different puborectalis configuration compared to controls. PMID- 17410576 TI - T1 measurement of flowing blood and arterial input function determination for quantitative 3D T1-weighted DCE-MRI. AB - PURPOSE: To propose a simple, accurate method for measuring T(1) in flowing blood and the arterial input function (AIF), and to evaluate the impact on dynamic contrast-enhanced MRI (DCE-MRI) quantification of pharmacokinetic parameters. MATERIALS AND METHODS: A total of 10 rabbits were scanned at 1.5 Tesla and administered a bolus of Gadomer. Preinjection T(1) and AIF measurements were acquired in the iliac arteries using a rapid three-dimensional (3D) spoiled gradient recalled echo (SPGR) approach. Correction was made for imperfect B(1) fields, in-flow, and partial volume effects. DCE-MRI parameters blood volume (v(b)) and endothelial transfer constant (K(trans)) in resting skeletal muscle were estimated from pharmacokinetic analysis using individually measured AIFs. Literature comparisons were made to assess accuracy. RESULTS: Blood T(1) was more accurate and precise after correction for B(1) and partial volume errors (1267 +/ 72 msec). Measured AIFs followed reported biexponential decay characteristics for Gadomer clearance in rabbits. Parameters v(b) (2.47 +/- 0.65%) and K(trans) (3.6 +/- 1.0 x 10(-3) minute(-1)) derived from AIFs based on corrected blood T(1)s were more reproducible and in better agreement with literature values. CONCLUSION: The proposed method enables accurate in vivo blood T(1) and AIF measurements and can be easily implemented in a range of DCE-MRI applications to improve both the accuracy and reproducibility of pharmacokinetic parameters. PMID- 17410577 TI - FAIR-TrueFISP imaging of cerebral perfusion in areas of high magnetic susceptibility differences at 1.5 and 3 Tesla. AB - PURPOSE: To estimate cerebral blood perfusion in areas of strong magnetic susceptibility changes with high spatial and temporal resolution using a flow sensitive alternating inversion recovery (FAIR) arterial spin labeling (ASL) method. MATERIALS AND METHODS: We implemented an ASL method that is capable of imaging perfusion in areas of high magnetic susceptibility changes by combining a FAIR spin preparation with a true fast imaging in steady precession (TrueFISP) data acquisition strategy. A TrueFISP readout sequence was applied especially in regions with magnetic field inhomogeneities and compared with corresponding FAIR measurements obtained with a standard echo-planar imaging (EPI) readout. Quantitative perfusion images were obtained at 1.5 Tesla (1.5T) from eight healthy volunteers (24-42 years old) and one patient (23 years old). FAIR TrueFISP perfusion images were compared with FAIR echo-planar images. T1 maps, which are necessary for quantitative perfusion estimation, were obtained with inversion recovery (IR) TrueFISP and IR EPI. Additionally, high-resolution perfusion measurements were performed on four volunteers at 3T. RESULTS: The two ASL perfusion imaging modalities yielded comparable diagnostic image quality in brain areas with low susceptibility differences at 1.5T. Cerebral perfusion of gray matter (GM) areas was 105.7 +/- 5.2 mL/100 g/minute for FAIR-TrueFISP and 88.8 +/- 14.6 mL/100 g/minute for FAIR-EPI at 1.5T, and 70.4 +/- 7.1 mL/100 g/minute for FAIR-TrueFISP and 63.5 +/- 6.9 mL/100 g/minute for FAIR-EPI at 3.0T. Higher perfusion values at 1.5T were due to more pronounced partial-volume effects from fast moving spins at lower spatial resolution. The FAIR-TrueFISP sequence showed no significant distortions and markedly reduced signal void artifacts in areas of high susceptibility changes (e.g., near brain-bone transitions and close to metallic clips) compared to FAIR-EPI. At 3T, highly resolved FAIR-TrueFISP perfusion images were acquired with an in-plane resolution of up to 1.3 mm. CONCLUSION: FAIR-TrueFISP allows for assessment of cerebral perfusion in areas of critically high susceptibility changes with conventional ASL methods. PMID- 17410578 TI - The H reflex from the abductor brevis hallucis muscle in healthy subjects. AB - In order to evaluate the clinical applicability of a neurophysiological technique, information is required about the effect of technical and subjective factors on the measured parameters, and whether these measures are reliable. The reliability of measurement of a given parameter is inversely related to the variability shown by a set of measurements of that parameter made on the same subject under similar conditions. Reliable measures are fundamental to the clinical utility of any technique. We evaluated the reliability using the intraclass correlation coefficient (ICC), and assessed both single-point and test retest normal limits for H-reflex parameters of abductor brevis hallucis (ABH). The H reflex from the ABH muscle was recorded in 36 of 43 subjects, but could not be elicited in 7 of 11 subjects who were >60 years of age. The Hmax latency increased as height increased, whereas Hmax amplitude and Hmax/Mmax amplitude ratio decreased as age and height increased. The Hmax latency and Hmax/Mmax amplitude ratio, but not Hmax amplitude, proved to be reliable. The usefulness of the H reflex from ABH in the evaluation of S2 radiculopathies is described. PMID- 17410579 TI - Autosomal-recessive and X-linked forms of hereditary motor and sensory neuropathy in childhood. AB - The hereditary motor and sensory neuropathies (HMSNs, Charcot-Marie-Tooth neuropathies) are the most common degenerative disorders of the peripheral nervous system. In recent years a dramatic expansion has occurred in our understanding of the molecular basis and cell biology of the recessively inherited demyelinating and axonal neuropathies, with delineation of a number of new neuropathies. Mutations in some genes cause a wide variety of clinical, neurophysiologic, and pathologic phenotypes, rendering diagnosis difficult. The X linked forms of HMSN represent at least 10%-15% of all HMSNs and have an expanded disease spectrum including demyelinating, intermediate, and axonal neuropathies, transient central nervous system (CNS) dysfunction, mental retardation, and hearing loss. This review presents an overview of the recessive and X-linked forms of HMSN observed in childhood, with particular reference to disease phenotype and neurophysiologic and pathologic abnormalities suggestive of specific diagnoses. These findings can be used by the clinician to formulate a differential diagnosis and guide targeted genetic testing. PMID- 17410580 TI - Structural variation manipulates the differential oxidative susceptibility and conformational stability of apolipoprotein E isoforms. AB - A growing amount of evidence implicates the involvement of apolipoprotein E (apoE) in the development of late-onset and sporadic forms of Alzheimer's disease (AD). It is now generally believed that the epsilon4 allele is associated with AD and the oxidative stress is more pronounced in AD. However, only limited data are available on apoE isoform-specificity and its relationship to both the oxidative susceptibility and conformational stability of apoE. In this article, we use site directed mutagenesis to investigate the structural role of amino acid residue 112, which is the only differing residue between apoE3 and E4. We examine the structural variation manipulating the oxidative susceptibility and conformational stability of apolipoprotein E isoforms. Arg112 in apoE4 was changed to Ala and Glu. Previous research has reported that apoE4 is more susceptible to free radicals than apoE3. In protein oxidation experiments, apoE4-R112A becomes more resistant to free radicals to the same extent as apoE3. In contrast, apoE4-R112E becomes the most susceptible protein to free radicals among all the apoE proteins. We also examine the conformational stability and the quaternary structural change by fluorescence spectroscopy and analytical ultracentrifugation, respectively. ApoE3 and E4 show apparent three- and two state unfolding patterns, respectively. ApoE4-R112A, similar to apoE3, demonstrates a biphasic denaturation with an intermediate that appears. The denaturation curve for apoE4-R112E, however, also displays a biphasic profile but with a slight shoulder at approximately 1.5M GdmCl, implying that an unstable intermediate existed in the denaturation equilibrium. The size distribution of apoE isoforms display similar patterns. ApoE4-R112E, however, has a greater tendency to dissociate from high-molecular-weight species to tetramers. These experimental data suggest that the amino acid residue 112 governs the differences in salt-bridges between these two isoforms and thus has a significant impact on the free radical susceptibility and structural variation of the apoE isoforms. PMID- 17410581 TI - Proton pathways and H+/Cl- stoichiometry in bacterial chloride transporters. AB - H+/Cl- antiport behavior has recently been observed in bacterial chloride channel homologs and eukaryotic CLC-family proteins. The detailed molecular-level mechanism driving the stoichiometric exchange is unknown. In the bacterial structure, experiments and modeling studies have identified two acidic residues, E148 and E203, as key sites along the proton pathway. The E148 residue is a major component of the fast gate, and it occupies a site crucial for both H+ and Cl- transport. E203 is located on the intracellular side of the protein; it is vital for H+, but not Cl-, transport. This suggests two independent ion transit pathways for H+ and Cl- on the intracellular side of the transporter. Previously, we utilized a new pore-searching algorithm, TransPath, to predict Cl- and H+ ion pathways in the bacterial ClC channel homolog, focusing on proton access from the extracellular solution. Here we employ the TransPath method and molecular dynamics simulations to explore H+ pathways linking E148 and E203 in the presence of Cl- ions located at the experimentally observed binding sites in the pore. A conclusion is that Cl- ions are required at both the intracellular (S(int)) and central (S(cen)) binding sites in order to create an electrostatically favorable H+ pathway linking E148 and E203; this electrostatic coupling is likely related to the observed 1H+/2Cl- stoichiometry of the antiporter. In addition, we suggest that a tyrosine residue side chain (Y445), located near the Cl- ion binding site at S(cen), is involved in proton transport between E148 and E203. PMID- 17410582 TI - Crystal structure of a binary complex between human GCN5 histone acetyltransferase domain and acetyl coenzyme A. PMID- 17410583 TI - Lactate, choline, and creatine levels measured by vitro 1H-MRS as prognostic parameters in patients with non-small-cell lung cancer. AB - PURPOSE: To determine the biochemical characteristics of lung cancer tissue using in vitro (1)H-MRS, and investigate the correlation between survival probabilities and lactate (Lac), creatine (Cr), and choline (Cho) concentrations measured by in vitro (1)H-MRS. MATERIALS AND METHODS: A total of 21 patients with lung cancer were included in this retrospective study. (1)H-MRS spectra measurements were performed at 6.35T using a JNM-EX270, high-resolution FT-NMR spectrometer. RESULTS: When normal lung tissue was compared with lung cancer tissue, significant differences were noted most consistently in the levels of Lac and Cho, with lung cancer tissue showing higher values than normal lung tissue. Lac concentrations of lung cancer tissue were significantly higher in patients with recurrence compared to patients without recurrence (0.285 +/- 0.096 mumol/g). The mean overall survival of patients in the low-Lac group was 50.28 +/- 6.47 months, which is significantly higher compared to the high-Lac group, which had a mean survival time of only 30.49 +/- 5.41 months. CONCLUSION: Kaplan-Meier analysis of the data showed that the overall and disease-free survival probabilities were significantly higher in patients with low tumor Lac values than in those with high tumor Lac concentrations. PMID- 17410584 TI - Postnatal evaluation of schizencephaly by transfontanellar three-dimensional sonography. AB - Schizencephaly is a rare anomaly of neuronal migration characterized by the presence of brain clefts that communicate with the lateral ventricles. Type I is characterized by clefts with fused lips or margins, not communicating with the subarachnoid space. Type II is characterized by longer clefts that communicate with the subarachnoid space. Neonatal diagnosis of schizencephaly on transfontanellar two-dimensional (2D) sonography is rare, with only 1 report in the medical literature. The major limitation of 2D sonography is its inability to assess neonatal prognosis. There are no reports on MEDLINE about the use of transfontanellar three-dimensional (3D) sonography in the assessment of schizencephaly. We present a case of type II schizencephaly diagnosed on the 29th week of gestation with 2D sonography and describe the main findings with 3D sonography in surface and transparency modes performed in the neonatal period via the fontanel. PMID- 17410585 TI - Sonographic diagnosis of uncomplicated first-trimester pregnancy in the rudimentary horn of a unicornuate uterus. AB - Although pregnancy in a rudimentary uterine horn is rare, the correct diagnosis of this condition on routine sonographic examination is critical, because its natural history usually involves the rupture of the pregnant horn during the second or third trimester, resulting in life-threatening bleeding. We report a case in which the diagnosis of right unicornuate uterus with a left noncommunicating rudimentary horn was made in the first trimester on routine sonographic examination. PMID- 17410586 TI - Severe form of congenital cerebral and cerebellar atrophy: a neurodegenerative disorder of fetal onset. AB - Infantile olivopontocerebellar atrophy (OPCA) is a rare congenital disorder likely due to an intrauterine neurodegenerative condition. Characteristic presentations are failure to thrive, cerebellar ataxia, respiratory insufficiency, and hypotonia or hypertonia. A few cases with severe manifestations (eg, the Pena-Shokeir phenotype) presenting in the neonatal period have also been reported. We present a case of infantile OPCA with the Pena Shokeir II phenotype and severe atrophy of the cerebellum and cerebral hemispheres. Comparison of prenatal sonographic findings of the fetal brain at 30 weeks' menstrual age and CT findings during the neonatal period indicated prenatal onset of the neurodegenerative process, which progressed rapidly during the last trimester. PMID- 17410587 TI - The dynamic fetal brain. AB - PURPOSE: To evaluate fetuses with normal intracranial anatomy in the second trimester that became abnormal in the third trimester. METHODS: We sonographically examined 6 fetuses with a normal second-trimester head sonogram that presented later in pregnancy with an abnormal head sonogram. RESULTS: Four categories of intracranial pathology were depicted: obstructive hydrocephalus, intraventricular intracranial hemorrhage, non-intraventricular intracranial hemorrhage, and porencephaly. CONCLUSIONS: Despite a normal midtrimester intracranial examination, evaluation of the fetal intracranial contents should be undertaken in subsequent sonographic examinations, because significant pathology can develop spontaneously. PMID- 17410588 TI - First-trimester sonography: is the fetus exposed to high levels of acoustic energy? AB - PURPOSE: As a form of energy, diagnostic ultrasound has bioeffects on living tissues. The thermal index (TI), TIS (TI for soft tissue), TIB (TI for bone), TIC (TI for cranial bone) expresses the potential for rise in temperature at the ultrasound beam's focal point. The mechanical index (MI) indicates the potential for the ultrasound beam to induce inertial cavitation in tissues. The goal of this study was to characterize the acoustic output of clinical ultrasound instruments, as expressed by TI and MI, during routine first-trimester sonographic examinations. METHODS: A prospective observational study was conducted. First-trimester patients were randomly selected from those scheduled for viability scans. An obstetrician collected data. Sonographers were blinded to the data being sought, which included gestational age, duration of the examination, and every variation in the MI and TI during each sonographic examination. RESULTS: A total of 52 first-trimester examinations were evaluated. The mean gestational age was 8.9 +/- 1.9 weeks. The mean duration of the sonographic examinations was 8.1+/- 1.4 minutes. During the examinations, there were 178 MI variations (mean +/- SD, 0.9 +/- 0.3) and 167 TI variations (mean +/- SD, 0.2 +/- 0.1). CONCLUSION: First-trimester sonographic examinations are associated with a negligible rise in TI. PMID- 17410589 TI - Short humerus: an additional antenatal sonographic feature of OFDS type II. AB - The prevalence of skeletal dysplasia in the neonatal period is 2.4/10,000 births; it accounts for 1-2% of perinatal deaths, mainly stillbirths. A long bone measurement of less than -4 standard deviations is a definitive indication of skeletal dysplasia, whereas measurement between -2 and -4 standard deviations warrants detailed sonographic examination for other associated anomalies to arrive at a diagnosis. We report a case of oral-facial-digital syndrome type II that presented antenatally with polyhydramnios, short humerus, polydactyly, and duplicated thumb. PMID- 17410590 TI - Acromioclavicular joint injury: enhanced technique of examination with dynamic maneuver. AB - Acromioclavicular (AC) sprains can be graded in 3 to 6 different types according to Tossy or Rockwell, respectively. In mild sprains (Tossy and Rockwell I), movements in the AC joint are minimal, because the coraco-clavicular ligaments are intact. In these patients, stress radiography is usually normal, and sonographic examination at rest can be normal as well, showing minimal or no displacement between the 2 extremities of the bones. We present a simple dynamic maneuver to enhance the diagnosis of these mild sprains known as the cross-arm maneuver, in which the hand is placed on the opposite shoulder. The dynamic sonographic examination during this maneuver clearly shows abnormal movements in the clavicle's extremity, which "falls down" to the acromion in the cross-arm position and is raised and pulled from the acromion at rest. The maneuver is very easy to perform and may be useful when a mild AC joint sprain is suspected. PMID- 17410591 TI - Electrodiagnostic studies of the facial nerve in peripheral facial palsy and hemifacial spasm. AB - Electrodiagnostic (EDX) assessment is one of the most important aspects in the evaluation of the two most common disorders of the facial nerve: facial palsy and hemifacial spasm. Facial palsy is usually an acute disorder that resolves in a few weeks but, in a number of cases, leads to a postparalytic facial syndrome featuring muscle synkinesis, myokymia, and involuntary mass contractions of muscles on the affected side. Hemifacial spasm is usually a chronic disorder characterized by paroxysms of involuntary, clonic, and synchronous twitching of all facial muscles on the affected side. EDX studies provide information on lesion location and severity, pathophysiology underlying the two disorders, and differential diagnosis between syndromes presenting with abnormal facial muscle activity. This monograph is intended to describe the most relevant EDX findings in the two disorders and the most appropriate timing for the examinations in order to provide useful information for prognosis and therapeutic decision making. PMID- 17410592 TI - Matrix metalloproteinases in neuromuscular disease. AB - Matrix metalloproteinases (MMPs), a family of zinc-dependent endoproteinases, are effector molecules in the breakdown of the blood-brain and blood-nerve barrier, and promote neural tissue invasion by leukocytes in inflammatory diseases of the central and peripheral nervous systems. Moreover, MMPs play an important role in synaptic remodeling, neuronal regeneration, and remyelination. Recent work concerning MMPs in patients with neuropathy, myopathy, spinal cord injury, and amyotrophic lateral sclerosis (ALS), and in corresponding animal models, is discussed in this review. PMID- 17410593 TI - Intraoperative muscle measurements reveal a relationship between contracture formation and muscle remodeling. AB - Children with cerebral palsy often have spasticity of both the extensors and flexors, but how and why a flexion contracture of the wrist will develop during growth is not thoroughly understood. In order to understand the muscle adaptations that occur during contracture formation, the relationship between intraoperative sarcomere length and the extent of contracture was measured in 23 children (average age, 14.3 +/- 2.9 years) undergoing tendon transfers involving the flexor carpi ulnaris (FCU) or extensor carpi radialis brevis (ECRB) muscles. For both ECRB and FCU, sarcomere lengths measured intraoperatively were longer compared to sarcomere lengths predicted from a regression relationship obtained from "control" patients with radial nerve injury (P < 0.001). The most interesting aspect of the long FCU sarcomere lengths measured was that there was a highly significant correlation between the degree of contracture formation and intraoperative sarcomere length (r2 = 0.5, P < 0.005). These data clearly show that greater contracture severity is associated with longer intraoperative FCU sarcomere length. No such correlation was observed for the ECRB. The data suggest that the clinical presentation of the contracture is related to degree of FCU, but not ECRB adaptation, to the central nervous system lesion. PMID- 17410594 TI - Facile production of mono-substituted urea side chains in solid phase peptide synthesis. AB - A method is reported for the straightforward generation of urea-containing peptides during Boc solid phase peptide synthesis. Primary amine side chains are converted to mono-alkyl ureas in two steps via an intermediate p-nitrophenyl carbamate. Use of p-methoxybenzyl amine as an ammonia equivalent affords mono alkyl final products from standard resin cleavage methods, without the need for additional steps. The reaction is highly efficient and applicable to variable length side chains and peptides. PMID- 17410595 TI - Solution structure of a novel D-naphthylalanine substituted peptide with potential antibacterial and antifungal activities. AB - A new type of Trp-rich peptide, Ac-KWRRWVRWI-NH2, designated as Pac-525, was found to possess improved activity against both gram-positive and negative bacteria. We have synthesized two Pac-525 analogues, D-Pac-525 containing all D amino acids and D-Nal-Pac-525, the D-Pac-525 analogue with tryptophan replaced by D-beta-naphthylalanine. We have determined the solution structure of D-Nal-Pac 525 bound to membrane-mimetic DPC micelles by two-dimensional NMR methods. The DPC micelle-bound structure of D-Nal-Pac-525 adopts a left-hand alpha-helical segment and the positively charged residues are clustered together to form a hydrophilic patch. The surface electrostatic potential map indicates the three D beta-naphthylalanines are packed against the peptide backbone and form an amphipathic structure. A variety of biophysical and biochemical experiments, including circular dichroism, fluorescence spectroscopy, and microcalorimetry, were used to show that D-Nal-Pac-525 interacted strongly with negatively charged phospholipid vesicles and induced efficient dye release from these vesicles, suggesting that the strong antimicrobial activity of D-Nal-Pac-525 may be due to interactions with bacterial and fungus membranes. PMID- 17410596 TI - Insulin structure and function. AB - Throughout much of the last century insulin served a central role in the advancement of peptide chemistry, pharmacology, cell signaling and structural biology. These discoveries have provided a steadily improved quantity and quality of life for those afflicted with diabetes. The collective work serves as a foundation for the development of insulin analogs and mimetics capable of providing more tailored therapy. Advancements in patient care have been paced by breakthroughs in core technologies, such as semisynthesis, high performance chromatography, rDNA-biosynthesis and formulation sciences. How the structural and conformational dynamics of this endocrine hormone elicit its biological response remains a vigorous area of study. Numerous insulin analogs have served to coordinate structural biology and biochemical signaling to provide a first level understanding of insulin action. The introduction of broad chemical diversity to the study of insulin has been limited by the inefficiency in total chemical synthesis, and the inherent limitations in rDNA-biosynthesis and semisynthetic approaches. The goals of continued investigation remain the delivery of insulin therapy where glycemic control is more precise and hypoglycemic liability is minimized. Additional objectives for medicinal chemists are the identification of superagonists and insulins more suitable for non injectable delivery. The historical advancements in the synthesis of insulin analogs by multiple methods is reviewed with the specific structural elements of critical importance being highlighted. The functional refinement of this hormone as directed to improved patient care with insulin analogs of more precise pharmacology is reported. PMID- 17410597 TI - Hepatic metastasectomy in children. AB - BACKGROUND: There are little data regarding the safety and efficacy of hepatic metastasectomy for solid tumors in childhood. We reviewed our institutional experience to assess operative mortality and morbidity, technique of resection, local control, and survival in pediatric patients undergoing liver resection for metastases. METHODS: All pediatric patients who underwent hepatic resection for metastatic disease from August 1988 to July 2005 were retrospectively identified and clinical data were collected. RESULTS: Fifteen patients were identified during this period and primary malignancies included neuroblastoma (7), Wilms tumor (3), osteogenic sarcoma (2), malignant gastric epithelial tumor (1), and desmoplastic small round cell tumor (2). Twelve patients underwent anatomical hepatic resections and 3 had wedge resections. There were no intraoperative or postoperative deaths. The 2 postoperative complications included 1 wound infection and 1 bile collection. The median follow-up after hepatic resection was 1.6 years (0.2-7 years). Three patients remain alive. Eleven patients died of progressive disease; 4 patients suffered local recurrence. One patient died from enterocolitis and sepsis and was without evidence of malignancy at the time of death. CONCLUSIONS: Hepatic metastasectomy in children is feasible and is associated with a low operative mortality and morbidity. In this small group of patients anatomic hepatectomy was associated with better local control compared with wedge resection. Overall prognosis in these patients remains poor and the decision to perform hepatic metastasectomy should be highly selective. PMID- 17410598 TI - Glutamine as an energy substrate in cultured neurons during glucose deprivation. AB - During glucose deprivation an increase in aspartate formation from glutamine has been observed in different brain preparations, including synaptosomes and cultured astrocytes. To what extent this reaction, which provides a substantial amount of energy, occurs in different types of neurons is unknown. The present study shows that (14)CO(2) formation from [U-(14)C]glutamine in cerebellar granule neurons, a glutamatergic preparation, increased by 60% during glucose deprivation, indicating enhanced aspartate formation or increased complete oxidative degradation of glutamine. In primary cultures of cerebrocortical interneurons, a GABAergic preparation, the rate of (14)CO(2) production from [U (14) C] glutamine was four times lower and not stimulated by glucose deprivation. During incubation with glutamine (0.8 mM) as the only metabolic substrate, cerebellar granule cells maintained an oxygen consumption rate of 12 nmol/min/mg protein, corresponding to an aspartate formation of 8 nmol/min/mg protein (three oxidations occur between glutamine and aspartate) or to a total oxidative degradation of 3 nmol/min/mg protein. During glucose deprivation, the rate of aspartate formation increased, and during a 20-min incubation in phosphate buffered saline it amounted to 3.3 nmol/min/mg protein at 0.2 mM glutamine, which might have been more if measured at 0.8 mM glutamine. These values are consistent with the rate of glutamine utilization calculated based on oxygen consumption and leaves open the possibility that some glutamine is completely degraded oxidatively, as has been shown by other authors based on pyruvate recycling and labeling of lactate from aspartate in cerebellar granule neurons. PMID- 17410599 TI - 24S-hydroxycholesterol in relation to disease manifestations of acute experimental autoimmune encephalomyelitis. AB - Levels of the brain-specific cholesterol metabolite 24S-hydroxycholesterol are proposed as possible biomarkers for multiple sclerosis (MS). It is not yet clear for which aspect of the MS disease manifestations 24S-hydroxycholesterol is a reflection. We studied the relation of serum levels of 24S-hydroxycholesterol and other sterols to the disease characteristics of acute experimental autoimmune encephalomyelitis (EAE), an animal model for MS. Serum was analyzed for cholesterol precursors, oxysterols, and plant sterols during the course of disease development. Significantly increased levels of the cholesterol metabolites 24S-hydroxycholesterol and 27-hydroxycholesterol were observed on day 9, before the onset of clinical signs. The serum levels of these oxysterols gradually increased up to 193% and 415%, respectively, at day 17, when clinical symptoms had recovered. Total cholesterol levels were slightly but significantly decreased on day 9 and day 17 in treated animals. Serum levels of cholesterol precursors and plant sterols decreased gradually from day 11 and day 14, respectively. Immunostaining of the 24S-hydroxycholesterol-forming enzyme Cyp46 was shown in macrophage infiltrates. In vitro experiments confirmed the presence of Cyp46 in macrophages and showed a decreased expression after LPS treatment. The data indicate that changes in serum oxysterols occur early in EAE and can be formed by macrophages. These early changes indicate an important role for oxysterols in the development of EAE. PMID- 17410600 TI - Role of c-Jun N-terminal kinase in early brain injury after subarachnoid hemorrhage. AB - The c-Jun N-terminal kinase (JNK) is induced by cerebral ischemia and injurious blood components acutely after subarachnoid hemorrhage (SAH). We hypothesized that inhibition of JNK will prevent damage to the neurovascular unit in the early brain injury period after SAH. Ninety-nine male SD rats (300-350 g) were randomly assigned to sham, SAH, and SAH treated with JNK inhibitor groups. SAH was induced by endovascular perforation. The JNK inhibitor SP600125 was administered intraperitoneally at 1 hr before and 6 hr after SAH. At 24 hr after SAH, we observed increased phosphorylation of JNK and c-Jun. Signs of neurovascular damage were observed in the hemorrhagic brains; these included the increases of aquaporin (AQP)-1 expression and brain water content as well as enhanced matrix metalloproteinase (MMP)-9 activity, vascular collagen IV loss, increased VEGF tissue level, and Evans blue extravasation. The appearances of cleaved caspase-3 expression, TUNEL-positive cells, and apoptotic morphology in cerebral tissues were associated with neurological deficit after SAH. JNK inhibition prevented c Jun phosphorylation and suppressed AQP1, MMP-9, VEGF, and caspase-3 activation, with concomitant diminution of neuronal injury, blood-brain barrier preservation, reduced brain swelling, and improved neurological deficit in rats after SAH. This study demonstrates a multitude of beneficial effects of JNK inhibition, including protection of the neurovascular unit in early brain injury after SAH. PMID- 17410601 TI - Glucose and hippocampal neuronal excitability: role of ATP-sensitive potassium channels. AB - Hyperglycemia-related neuronal excitability and epileptic seizures are not uncommon in clinical practice. However, their underlying mechanism remains elusive. ATP-sensitive K(+) (K(ATP)) channels are found in many excitable cells, including cardiac myocytes, pancreatic beta cells, and neurons. These channels provide a link between the electrical activity of cell membranes and cellular metabolism. We investigated the effects of higher extracellular glucose on hippocampal K(ATP) channel activities and neuronal excitability. The cell attached patch-clamp configuration on cultured hippocampal cells and a novel multielectrode recording system on hippocampal slices were employed. In addition, a simulation modeling hippocampal CA3 pyramidal neurons (Pinsky-Rinzel model) was analyzed to investigate the role of K(ATP) channels in the firing of simulated action potentials. We found that incremental extracellular glucose could attenuate the activities of hippocampal K(ATP) channels. The effect was concentration dependent and involved mainly in open probabilities, not single channel conductance. Additionally, higher levels of extracellular glucose could enhance neuropropagation; this could be attenuated by diazoxide, a K(ATP) channel agonist. In simulations, high levels of intracellular ATP, used to mimic increased extracellular glucose or reduced conductance of K(ATP) channels, enhanced the firing of action potentials in model neurons. The stochastic increases in intracellular ATP levels also demonstrated an irregular and clustered neuronal firing pattern. This phenomenon of K(ATP) channel attenuation could be one of the underlying mechanisms of glucose-related neuronal hyperexcitability and propagation. PMID- 17410602 TI - Axonal transport of neuropeptides: Retrograde tracing study in live cell cultures of rat sympathetic cervical ganglia. AB - Previous studies demonstrated that neuropeptides are transported with fast axonal transport. Considerable amounts (30-40%) of anterogradely transported peptides accumulated distal to a crush, apparently recycling to the cell bodies. In the present study, we used primary and compartmented cultures of sympathetic cervical ganglia (SCG) to address questions on the origin of the recycling peptides. In primary cultures, distinct labeling of neuropeptide Y (NPY) or secretoneurin (SN) immunoreactivities was detected in varicosities and in cell bodies, after administration of NPY or SN antibodies to the living cultures. Simultaneous addition to the medium with antibody against the N-terminal (lumen) domain of synaptotagmin, resulted in a partial overlapping between synaptotagmin and NPY/SN. In compartmented chamber cultures, in which cell body and proximal segments of the processes are restricted to the central chamber and the distal processes are present in peripheral compartments, antibody administration was performed in the peripheral compartment. KCl (60-120 mM) was added to the central chamber for 10 sec, followed by washing, and 30-60 min later clear labeling was detected in the cell bodies, suggesting that the antibodies were now present in structures that were transported from the distal segments in the peripheral compartment to the cell body. The results indicate 1) that peptide release from large dense cored vesicles is incomplete; 2) that the remaining peptides, together with the membrane, are retrogradely transported to cell bodies; and 3) that the recycling peptides accumulating distal to a crush of a peripheral nerve are most likely to be recycled from the nerve terminals. PMID- 17410603 TI - Effects of polyethylene glycol and magnesium sulfate administration on clinically relevant neurological outcomes after spinal cord injury in the rat. AB - The purpose of this study was to determine the long-term effects of polyethylene glycol (PEG) and magnesium sulfate (MgSO(4)) on clinically relevant motor, sensory, and autonomic outcomes after spinal cord injury (SCI). Rats were injured by clip compression (50 g; T4) and treated 15 min and 6 hr postinjury intravenously (tail vein) with PEG (1 g/kg, 30% w/w in saline; n = 11), MgSO(4) (300 mg/kg; n = 5), PEG + MgSO(4) (n = 6), or saline (n = 10). Behavioral testing lasted for 6 weeks, followed by histological analysis of the spinal cord. Both PEG and MgSO(4) resulted in enhanced locomotor recovery and lower susceptibility to neuropathic pain (mechanical allodynia) compared with saline. At 6 weeks, BBB scores were 7.3 +/- 0.2, 7.7 +/- 0.4, and 6.4 +/- 0.6 in PEG-treated, MgSO(4) treated, and saline-treated control groups, respectively. Likewise, at 6 weeks PEG-, MgSO(4)-, and saline-treated control animals showed 3.5 +/- 0.4, 2.8 +/- 0.9, and 5.0 +/- 0.5 avoidance responses to at-level touch, respectively. PEG + MgSO(4) improved locomotor recovery and reduced pain but did not provide additional benefit compared with either treatment alone. Neither treatment, nor their combination, attenuated mean arterial pressure (MAP) increases during autonomic dysreflexia. However, saline-treated controls had significantly lower resting MAP than PEG-treated rats and tended to have lower resting MAP than MgSO(4)-treated rats 6 weeks postinjury. MgSO(4) treatment and PEG + MgSO(4) treatment resulted in significant increases in dorsal myelin sparing, and the latter resulted in significant reductions in lesion volume, compared with saline treated controls. Furthermore, mean lesion volumes correlated negatively with the corresponding mean BBB scores and positively with the corresponding mean pain scores. In conclusion, both PEG and MgSO(4) enhanced long-term clinical outcomes after SCI. PMID- 17410604 TI - In vitro effects of 2-methoxyestradiol on cell morphology and Cdc2 kinase activity in SNO oesophageal carcinoma cells. AB - The effects of 1 x 10(-6) M exogenous 2-methoxyestradiol (2 ME) were determined on cell morphology and cell division cycle (Cdc) 2 kinase activity in SNO oesophageal carcinoma cells. Mitotic indices revealed an increase in metaphase cells (11.2%) when compared to the 0.5% vehicle-treated cells after 18 h of exposure to 2 ME. Vehicle-treated control cells did not show any hallmarks of apoptosis after 18 h of exposure to dimethyl sulphoxide. Only 0.5% of 2 ME treated cells showed characteristics of apoptosis. Conversely, increased morphological hallmarks of apoptosis were observed in SNO-treated cells after 21.5 h of 2 ME exposure. When compared to the 0.5% in vehicle-treated cells, 4.7% of cells were in apoptosis. Furthermore, 34.1% of cells were blocked in metaphase after 21.5 h of 2 ME exposure compared to 0.6% of vehicle-control cells. In addition, Cdc2 kinase activity was statistically significantly increased (1.3 fold) (p<0.005) in 2 ME-treated cells when compared to vehicle-treated controls. The present preliminary study suggests that the accumulation observed in metaphase cells and the increase in Cdc2 kinase activity caused by 2 ME are consistent with morphological hallmarks of mitotic arrest and disrupted mitotic spindle formation, thus leading to induction of apoptosis in SNO cells. PMID- 17410605 TI - Determination of midazolam and its hydroxy metabolites in human plasma and oral fluid by liquid chromatography/electrospray ionization ion trap tandem mass spectrometry. AB - Midazolam (MDZ), a short-acting benzodiazepine, is a widely accepted probe drug for CYP3A phenotyping. Published methods for its analysis have used either therapeutic doses of MDZ, or, if employing lower doses, were mostly unable to quantify the two hydroxy metabolites. In the present study, a sensitive and specific liquid chromatography/electrospray ionization tandem mass spectrometry method was developed and validated for the quantitative determination of MDZ and two of its metabolites (1'-hydroxymidazolam (1'-OHMDZ) and 4-hydroxymidazolam (4 OHMDZ)) in human plasma and oral fluid. After liquid-liquid extraction with hexane/dichloromethane (73:27, v/v), the analytes were separated on a Luna C18(2) (100 x 2.1 mm) analytical column using gradient elution. Detection was achieved using tandem mass spectrometry on an ion trap mass spectrometer. Midazolam-d6 was used as internal standard for quantification. The calibration curves were linear (R2 >0.998) between 0.05 and 20 ng/mL for MDZ and both metabolites in both matrices. Using 1 mL samples, the limit of detection was 0.025 ng/mL and the limit of quantification was 0.05 ng/mL for MDZ and the hydroxy metabolites in both matrices. Intra- and inter-day accuracies, determined at three different concentrations, were between 92.1 and 102.3% and the corresponding coefficients of variation were <7.3%. The average recoveries were 90.6%, 86.7% and 79.0% for MDZ, 1'-OHMDZ and 4-OHMDZ in plasma and 95.3%, 96.6% and 86.8% for MDZ, 1'-OHMDZ and 4-OHMDZ, respectively, in oral fluid. The method was successfully applied to a pharmacokinetic study, showing that MDZ and its hydroxy metabolites can be determined precisely in in vivo samples obtained following a single oral or intravenous dose of 2 mg MDZ. The method appears to be useful for CYP3A phenotyping in plasma using sub-therapeutic MDZ doses, but larger studies are needed to test this assumption. PMID- 17410609 TI - Lentivirus vector-mediated gene transfer to the developing bronchiolar airway epithelium in the fetal lamb. AB - BACKGROUND: Development of effective and durable gene therapy for treatment of the respiratory manifestations of cystic fibrosis remains a formidable challenge. Obstacles include difficulty in achieving efficient gene transfer to mature airway epithelium and the need to stably transduce self-renewing epithelial progenitor cells in order to avoid loss of transgene expression through epithelial turnover. Targeting the developing airway epithelium during fetal life offers the prospect of circumventing these challenges. METHODS: In the current study we investigated vesicular stomatitis virus glycoprotein (VSVg)-pseudotyped HIV-1-derived lentivirus vector-mediated gene transfer to the airway epithelium of mid-gestation fetal lambs, both in vitro and in vivo. In the in vitro studies epithelial sheet explants and lung organ culture were used to examine transduction of the proximal and more distal airway epithelium, respectively. For the in vivo studies, vector was delivered directly into the proximal airway. RESULTS: We found that even during the early pseudoglandular and canalicular phases of lung development, occurring through mid-gestation, the proximal bronchial airway epithelium was relatively mature and highly resistant to lentivirus-mediated transduction. In contrast, the more distal bronchiolar airway epithelium was relatively permissive for transduction although the absolute levels achieved remained low. CONCLUSION: This result is promising as the bronchiolar airway epithelium is a major site of pathology in the cystic fibrosis airway, and much higher levels of transduction are likely to be achieved by developing strategies that increase the amount of vector reaching the more distal airway after intratracheal delivery. PMID- 17410611 TI - Palladium complexes of N-heterocyclic carbenes as catalysts for cross-coupling reactions--a synthetic chemist's perspective. AB - Palladium-catalyzed C-C and C-N bond-forming reactions are among the most versatile and powerful synthetic methods. For the last 15 years, N-heterocyclic carbenes (NHCs) have enjoyed increasing popularity as ligands in Pd-mediated cross-coupling and related transformations because of their superior performance compared to the more traditional tertiary phosphanes. The strong sigma-electron donating ability of NHCs renders oxidative insertion even in challenging substrates facile, while their steric bulk and particular topology is responsible for fast reductive elimination. The strong Pd-NHC bonds contribute to the high stability of the active species, even at low ligand/Pd ratios and high temperatures. With a number of commercially available, stable, user-friendly, and powerful NHC-Pd precatalysts, the goal of a universal cross-coupling catalyst is within reach. This Review discusses the basics of Pd-NHC chemistry to understand the peculiarities of these catalysts and then gives a critical discussion on their application in C-C and C-N cross-coupling as well as carbopalladation reactions. PMID- 17410612 TI - Live attenuated Salmonella as a vector for oral cytokine gene therapy in melanoma. AB - Systemic administration of cytokines has shown therapeutic benefits in cancer patients; however, serious adverse effects associated with direct protein administration prevent the wide use of this approach. We have assessed the capacity of live attenuated Salmonella to act as a vector for oral cytokine-gene therapy. Salmonella orally administered to melanoma-bearing mice was found to accumulate within the tumor, reaching up to 10(5) bacteria per gram of tumor by day 21 after bacterial inoculation. Numbers of bacteria recovered from tumor did not differ from those recovered from liver or spleen at any time point. Recombinant bacteria carrying eukaryotic expression vectors encoding the murine IL-4 or IL-18 genes were administered to groups of mice with established subcutaneous melanoma tumors. We found that a single oral dose of Salmonella carrying any of the cytokine-encoding plasmids resulted in significantly increased survival time, as compared with mice that received Salmonella carrying the parental plasmid or PBS. Increased levels of IFNgamma were found in sera of animals receiving either of the cytokine-encoding bacteria, but not in mice receiving Salmonella alone or PBS. Co-administration of both recombinant bacteria maximized the production of IFNgamma. Overall these results suggest that cytokine encoding Salmonella can be an effective and safer alternative to systemic administration of cytokines for immunotherapy of cancer. PMID- 17410613 TI - Electroporation enhances reporter gene expression following delivery of naked plasmid DNA to the lung. AB - BACKGROUND: Existing methods of non-viral airway gene transfer suffer from low levels of efficiency. Electroporation has been used to enhance gene transfer in a range of tissues. Here we assess the usefulness of electroporation for enhancing gene transfer in the lungs of mice and sheep. METHODS: Naked plasmid DNA (pDNA) expressing either luciferase or green fluorescent protein (GFP) was delivered to mouse lungs by instillation. Following surgical visualisation, the lungs were directly electroporated and the level and duration of luciferase activity was assessed and cell types that were positive for GFP were identified in lung cryosections. Naked pDNA was nebulised to the sheep lung and electrodes attached to the tip of a bronchoscope were used to electroporate airway segment bifurcations, Luciferase activity was assessed in electroporated and control non electroporated regions, after 24 h. RESULTS: Following delivery of naked pDNA to the mouse lung, electroporation resulted in up to 400-fold higher luciferase activity than naked pDNA alone when luciferase was under the control of a cytomegalovirus (CMV) promoter. Following delivery of a plasmid containing the human polyubiquitin C (UbC) promoter, electroporation resulted in elevated luciferase activity for at least 28 days. Visualisation of GFP indicated that electroporation resulted in increased GFP detection compared with non electroporated controls. In the sheep lung electroporation of defined sites in the airways resulted in luciferase activity 100-fold greater than naked pDNA alone. CONCLUSIONS: These results indicate that electroporation can be used to enhance gene transfer in the lungs of mice and sheep without compromising the duration of expression. PMID- 17410614 TI - Polyplex-mediated gene transfer and cell cycle: effect of carrier on cellular uptake and intracellular kinetics, and significance of glycosaminoglycans. AB - BACKGROUND: Here we report on studies that probe whether the intracellular kinetics of plasmid DNA (pDNA) and cell surface glycosaminoglycans (GAGs) are modified during the cell cycle in a way that can be correlated with changes in gene transfer efficiency with poly(ethyleneimine) (PEI) and poly-L-lysine (PLL) polyplexes. METHODS: Synchronized D407 retinal cells were transfected with PEI and PLL polyplexes using a luciferase reporter. The free and/or loosely complexed nuclear pDNA was determined by real-time PCR, and compared with transgene expression, the rate of pinocytosis by FITC-dextran uptake and the content of cell surface GAGs. RESULTS: The amount of free and/or loosely complexed nuclear pDNA between cell cycle phases varied approximately 4-20 times (G1 < S < G2/M). Both carriers delivered pDNA in a similar way into the nucleus (PLL vs. PEI < or = 3.5-fold), but PEI was approximately 10-100 times more efficient in gene expression than PLL (G1 < G2/M < S). The rate of pinocytosis increased up to 70 fold from G1 to middle S phase. Cell surface heparan and chondroitin sulfate increased 50-80%, and hyaluronan decreased 50% when the cells went from G1 through S to G2/M. CONCLUSIONS: The data obtained indicates that no single parameter (pinocytosis, cell surface GAGs, nuclear uptake) solely accounts for the differential pDNA uptake or expression during cell cycle, and that the main difference in PLL- and PEI-mediated transfections seems to be at the nuclear level. PMID- 17410615 TI - Combined treatment with Ad-hTRAIL and DTIC or SAHA is associated with increased mitochondrial-mediated apoptosis in human melanoma cell lines. AB - BACKGROUND: Currently, dacarbazine (DTIC) is the only approved systemic treatment for metastatic malignant melanoma. However, the modest treatment effect encourages studies on novel therapeutic molecules, delivery systems and combination therapies. Full-length TRAIL, delivered from an adenoviral vector (Ad hTRAIL), was studied in combination with DTIC or the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) in human melanoma cell lines. METHODS: The cytotoxic potential of the combination treatments was assessed by cell viability measurements and CalcuSyn analysis. Involvement of apoptosis was analyzed by TUNEL staining, mitochondrial membrane potential measurements, and activation and expression levels of caspases and other mediators of apoptosis. RESULTS: Ad-hTRAIL in combination with DTIC or SAHA resulted in additive or synergistic growth inhibition compared to each treatment used as single agent. Both combinations augmented apoptosis, which was mediated through the death receptor (DR) pathway by enhanced activation of caspase-8, and through increased loss of mitochondrial integrity. Provoked cleavage of Bid, which bridges the extrinsic and intrinsic apoptosis pathways, and downregulation of the anti apoptotic mediators Bcl-X(L), Mcl-1 and XIAP (but not Bcl-2) were critical contributing factors. Increased levels of DR4 and DR5 were not a common underlying mechanism as DTIC did not affect the levels of either of the receptors. However, SAHA-induced expression of DR4 may have reduced the TRAIL resistance in the SKMEL-28 cell line. CONCLUSION: Administration of Ad-hTRAIL in combination with DTIC or SAHA enhances apoptosis in human melanoma cell lines, and suggests that the therapeutic potential of such treatment strategies should be further evaluated for possible clinical use. PMID- 17410616 TI - Desktop growth of carbon-nanotube monoliths with in situ optical imaging. PMID- 17410617 TI - Phase separation on mixed-monolayer-protected metal nanoparticles: a study by infrared spectroscopy and scanning tunneling microscopy. PMID- 17410618 TI - microRNA: an emerging therapeutic. PMID- 17410619 TI - Titanium-containing zeolites and microporous molecular sieves as photovoltaic solar cells. AB - Four titanium-containing zeolites and microporous molecular sieves differing on the crystal structure and particle size (Ti/Beta, Ti/Beta-60, TS-1 and ETS-10) are prepared, and their activity for solar cells after incorporating N3 (a commercially available ruthenium polypyridyl dye) is tested. All the zeolites exhibit photovoltaic activity, and the photoresponse is quite independent of the zeolite pore dimensions or particle size. The photoresponse increases with titanium content in the range 1-7% wt. In this way, cells are obtained that have open-circuit voltage Voc=560 mV and maximum short-circuit photocurrent density Isc=100 microA, measured for 1x1 cm2 surfaces with a solar simulator at 1000 W through and AM 1.5 filter. These values are promising and comparable to those obtained for current dye-sensitized titania solar cells. PMID- 17410620 TI - Water molecules adsorbed at electrode surfaces determine the macroscopic contact angles. PMID- 17410621 TI - Highly ordered nanostructured surfaces obtained with silica-filled diblock copolymer micelles as templates. AB - In this work, we present the size-controlled preparation of silica-filled micelle cores and their self-assembly behavior, which is dependent on the block lengths, different coating techniques, and substrates. Furthermore, we present a way to use these structures as templates for highly ordered magnetic nanostructures, revealed by Ar-ion milling. The resulting structures were characterized by different imaging and scattering techniques and model simulations were performed. The characterization of the obtained nanostructured surfaces has be performed with atomic force microscopy, by scanning electron microscopy, grazing-incidence X-ray small-angle scattering, and X-ray reflectivity measurements. The magneto optical Kerr effect was utilized to investigate magnetic properties. PMID- 17410622 TI - Structure induction of the T-cell receptor zeta-chain upon lipid binding investigated by NMR spectroscopy. AB - The conformation of the cytoplasmic part of the zeta-chain of the T-cell receptor (TCR) in its free form and bound to detergent micelles has been investigated by heteronuclear NMR spectroscopy. The zeta-chain is considered to be a mediator between the extracellular antigen and the intracellular signal-transduction cascade leading to T-cell activation. Earlier studies suggested a T-cell activation mechanism that involved a TCR-state-dependent lipid incorporation propensity of the zeta-chain accompanied by a helical folding transition. In order to support this proposed mechanism, standard protein NMR assignment and secondary-structure-elucidation techniques have been applied to the free TCR zeta chain and to the zeta-chain bound to the detergent LMPG, which forms a micelle, in order to obtain the structural characteristics of this folding transition in a residue-resolved manner. We could assign the resonances of the free zeta-chain at 278 K, and this formed the basis for chemical-shift-perturbation studies to identify lipid binding sites. Our NMR results show that the free TCR zeta-chain is indeed intrinsically unstructured. Regions around the ITAM2 and ITAM3 sequences are involved in a highly dynamic binding of the free zeta-chain to a detergent micelle formed by the acidic lipid LMPG. PMID- 17410623 TI - Simple and rapid colorimetric biosensors based on DNA aptamer and noncrosslinking gold nanoparticle aggregation. PMID- 17410624 TI - A novel approach towards carbon-Ru electrodes with mesoporosity for supercapacitors. PMID- 17410625 TI - Carbon-fluorine bond activation in fluoroolefins: clear documentation of cooperative C-F bond activation by adjacent metal centers. PMID- 17410626 TI - Helical microstructure of polynorbornene. PMID- 17410627 TI - S2-->S1 internal conversion in beta-carotene: strong vibronic coupling from amplitude oscillations of transient absorption bands. PMID- 17410628 TI - Near-infrared dyes and fluorophores based on diketopyrrolopyrroles. PMID- 17410629 TI - Molecular transmission: controlling the twist sense of a helical polymer with a single light-driven molecular motor. PMID- 17410630 TI - Racemic beta sheets in biochirogenesis. PMID- 17410631 TI - RuO2 clusters within LTA zeolite cages: consequences of encapsulation on catalytic reactivity and selectivity. PMID- 17410632 TI - Base dependence in copper-catalyzed Huisgen reactions: efficient formation of bistriazoles. PMID- 17410633 TI - 10a-aza-10b-borapyrenes: heterocyclic analogues of pyrene with internalized BN moieties. PMID- 17410634 TI - Involvement of morbilliviruses in the pathogenesis of demyelinating disease. AB - Two members of the morbillivirus genus of the family Paramyxoviridae, canine distemper virus (CDV) and measles virus (MV), are well-known for their ability to cause a chronic demyelinating disease of the CNS in their natural hosts, dogs and humans, respectively. Both viruses have been studied for their potential involvement in the neuropathogenesis of the human demyelinating disease multiple sclerosis (MS). Recently, three new members of the morbillivirus genus, phocine distemper virus (PDV), porpoise morbillivirus (PMV) and dolphin morbillivirus (DMV), have been discovered. These viruses have also been shown to induce multifocal demyelinating disease in infected animals. This review focuses on morbillivirus-induced neuropathologies with emphasis on aetiopathogenesis of CNS demyelination. The possible involvement of a morbillivirus in the pathogenesis of multiple sclerosis is discussed. PMID- 17410635 TI - Yield of a new method to detect cognitive impairment in general practice. AB - OBJECTIVE: To examine whether a new screening method that identified patients with cognitive impairment who needed further examination on the presence of dementia yielded patients who were not detected by their general practitioner (GP), and to identify factors associated with GPs' awareness of patients identified by the screening. METHODS: Cross-sectional comparison between two methods used to identify dementia symptoms: (1) usual identification of dementia by GPs; (2) a two-stage screening to identify cognitive impairment. The two methods were implemented on the same older general practice population. The study was set in primary care practices in the Netherlands. The participants were 44 GPs and 2,101 general practice patients aged 75+ who lived at home. The following measurements were used: (1) yield of the screening; (2) determinants of GPs' awareness of patients identified by the screening. RESULTS: The two-stage screening yielded 117 patients with cognitive impairment who needed further examination; in most cases (n = 82, 70.1%) their GP was unaware of the symptoms. Among patients identified by the screening, GPs' awareness was associated with co morbidity of chronic diseases [odds ratio (OR) = 3.19; 95% Confidence Interval (CI) = 1.25 to 8.15], depressive symptoms (OR = 0.41; 95% CI = 0.17 to 0.99), and cognitive functioning (per point on the MMSE, OR = 0.88; 95% CI = 0.79 to 0.98). CONCLUSION: A two-stage screening method and increased alertness for cognitive impairment and dementia among patients with depressive symptoms may improve detection rate of dementia in general practice. PMID- 17410636 TI - Hospital volume does not influence long-term survival of patients undergoing surgery for oesophageal or gastric cancer. AB - BACKGROUND: Guidelines suggest that surgery for oesophageal and gastric cancer should be conducted in large cancer centres. This national study examined the relationship between hospital volume and outcome in Scotland. METHODS: This was a prospective, population-based study of 3293 consecutive patients with oesophageal or gastric cancer diagnosed between 1997 and 1999. Some 1302 patients underwent surgery and were followed for 5 years after operation. RESULTS: The 5-year adjusted overall survival rate for the 3293 patients was 18.7 (95 per cent confidence interval (c.i.) 17.2 to 20.2) per cent and that after surgical resection was 39.6 (95 per cent c.i. 36.3 to 43.0) per cent. Death within 1 year after surgical resection was associated with a postoperative complication (odds ratio (OR) 2.5 (95 per cent c.i. 1.6 to 3.8); P < 0.001) or resection margin involvement by tumour (OR 7.2 (95 per cent c.i. 1.1 to 47.5); P = 0.042) after adjustment for age, sex and tumour location. There was no relationship between hospital volume and postoperative morbidity or mortality, nor between survival and volume of patients either for hospital of diagnosis or hospital of surgery. CONCLUSION: This population-based study of oesophageal and gastric cancer suggests that the link between hospital volume and long-term survival for patients undergoing surgery requires re-evaluation. PMID- 17410637 TI - Impact of surgeon volume and specialization on short-term outcomes in colorectal cancer surgery. AB - BACKGROUND: Several studies have shown a relationship between surgeon volume and outcomes in colorectal cancer surgery. The aim of this study was to determine the impact of surgeon volume and specialization on primary tumour resection rate, restoration of bowel continuity following rectal cancer resection, anastomotic leakage and perioperative mortality. METHODS: The Northern Region Colorectal Cancer Audit Group conducts a population-based audit of patients with colorectal cancer managed by surgeons. This study examined 8219 patients treated between 1998 and 2002. Outcomes were modelled using multivariate logistic regression analysis. RESULTS: Tumour resection was performed in 6949 (93.8 per cent) of 7411 patients. High-volume surgeons with an annual caseload of at least 18.5 (odds ratio (OR) 1.53 (95 per cent confidence interval (c.i.) 1.10 to 2.12); P = 0.012) and colorectal specialists (OR 1.42 (95 per cent c.i. 1.06 to 1.90); P = 0.018) were more likely to perform elective sphincter-saving rectal surgery. In elective surgery, the risk of perioperative death was lower for high-volume surgeons (OR 0.58 (95 per cent c.i. 0.44 to 0.76); P < 0.001), but this was not the case in emergency surgery. CONCLUSION: High-volume surgeons had lower perioperative mortality rates for elective surgery, and were more likely to use restorative rectal procedures. PMID- 17410638 TI - Isolated tumour cells in blood and E-cadherin expression in oesophageal squamous cell cancer. AB - BACKGROUND: Patients with oesophageal squamous cell carcinoma have a high rate of recurrence, even after curative resection. The aim of this study was to examine the correlation between the presence of isolated tumour cells (ITCs) in the blood and recurrence, and between the presence of ITCs and E-cadherin expression in the primary tumour in these patients. METHODS: Blood samples obtained immediately before and after resection in 125 patients with oesophageal squamous cell carcinoma were examined by real-time reverse transcription-polymerase chain reaction using carcinoembryonic antigen mRNA. Blood samples from 28 healthy volunteers and 42 patients with benign diseases were used as controls. RESULTS: Seventy-seven patients (61.6 per cent) were ITC positive. ITC positivity correlated significantly with tumour depth, lymph node metastasis, stage, lymphatic invasion and venous invasion. Multivariable analysis revealed that tumour depth and ITC positivity were independent factors for a shortened haematogenous disease-free interval. A significant correlation was found between ITC positivity and reduced E-cadherin expression in the primary tumour (P < 0.001). ITC-positive patients with preserved E-cadherin expression had a longer disease-free interval (P = 0.016), haematogenous disease-free interval (P = 0.020) and overall survival (P = 0.004) than those with reduced E-cadherin expression. CONCLUSION: Examination of ITCs in the blood is useful for predicting haematogenous recurrence in patients with oesophageal squamous cell carcinoma. PMID- 17410639 TI - Careers in cardiology: Germany. Interview with Dr. Hugo Katus by Barry Shurlock. PMID- 17410640 TI - The PIP5K2A and RGS4 genes are differentially associated with deficit and non deficit schizophrenia. AB - Several putative schizophrenia susceptibility genes have recently been reported, but it is not clear whether these genes are associated with schizophrenia in general or with specific disease subtypes. In a previous study, we found an association of the neuregulin 1 (NRG1) gene with non-deficit schizophrenia only. We now report an association study of four schizophrenia candidate genes in patients with and without deficit schizophrenia, which is characterized by severe and enduring negative symptoms. Single-nucleotide polymorphisms (SNPs) were genotyped in the DTNBP1 (dysbindin), G72/G30 and RGS4 genes, and the relatively unknown PIP5K2A gene, which is located in a region of linkage with both schizophrenia and bipolar disorder. The sample consisted of 273 Dutch schizophrenia patients, 146 of whom were diagnosed with deficit schizophrenia and 580 controls. The strongest evidence for association was found for the A-allele of SNP rs10828317 in the PIP5K2A gene, which was associated with both clinical subtypes (P = 0.0004 in the entire group; non-deficit P = 0.016, deficit P = 0.002). Interestingly, this SNP leads to a change in protein composition. In RGS4, the G-allele of the previously reported SNP RGS4-1 (single and as part of haplotypes with SNP RGS4-18) was associated with non-deficit schizophrenia (P = 0.03) but not with deficit schizophrenia (P = 0.79). SNPs in the DTNBP1 and G72/G30 genes were not significantly associated in any group. In conclusion, our data provide further evidence that specific genes may be involved in different schizophrenia subtypes and suggest that the PIP5K2A gene deserves further study as a general susceptibility gene for schizophrenia. PMID- 17410641 TI - Markedly attenuated acute and chronic pain responses in mice lacking adenylyl cyclase-5. AB - Chronic inflammatory and neuropathic pain is often difficult to manage using conventional remedies. The underlying mechanisms and therapeutic strategies required for the management of chronic pain need to be urgently established. The cyclic AMP (cAMP) second messenger system has been implicated in the mechanism of nociception, and the inhibition of the cAMP pathway by blocking the activities of adenylyl cyclase (AC) and protein kinase A has been found to prevent chronic pain in animal models. However, little is known regarding which of the 10 known isoforms of AC are involved in nociceptive pathways. Therefore, we investigated the potential pronociceptive function of AC5 in nociception using recently developed AC5 knockout mice (AC5-/-). We found that AC5-/- mice show markedly attenuated pain-like responses in acute thermal and mechanical pain tests as compared with the wildtype control. Also, AC5-/- mice display hypoalgesic responses to inflammatory pain induced by subcutaneous formalin injection into hindpaws, and to non-inflammatory and inflammatory visceral pain induced by injecting magnesium sulfate or acetic acid into the abdomen. Moreover, AC5-/- mice show strongly suppressed mechanical and thermal allodynia in two nerve injury-induced neuropathic pain models. These results suggest that AC5 is essential for acute and chronic pain, and that AC5 knockout mice provide a useful model for the evaluation of the pathophysiological mechanisms of pain. PMID- 17410642 TI - [Carcinosarcoma of the larynx and hypopharynx]. AB - INTRODUCTION: A carcinosarcoma is an uncommon neoplasm with both malignant epithelial and malignant mesenchymal components. The tumor entity has been described in many epithelial organs. Manifestation in the larynx and the hypopharynx is rare. PATIENTS: This report recounts two patients, one with a carcinosarcoma of the vocal cords and the other with a hypopharynx tumour. On the background of the literature we discuss the peculiarities of histology, pathogenesis, clinical features and therapy. DISCUSSION: Carcinosarcomas of the larynx and hypopharynx are typically described in elderly men. Smoking, alcohol and previous irradiation of the head and neck are predisposing etiologic factors. Clinical symptoms correspond to other neoplasms of the area--a short anamnesis is typical. In most cases the endoscopy reveals the presence of an exophytic lesion with intact epithelial surface. Crucial for the diagnosis is the revelation of a carcinoma together with sarcoma in the histology and immunohistochemistry both in the primary and secondary side. The histogenesis of carcinosarcoma is not yet clear--it seems that a pluripotent cell produces both tumour components. Most authors recommend complete surgical excision of the primary, regional lymphadenectomy and postoperative irradiation. Despite of an often large tumor volume the preservation of the larynx seems to be possible in many cases, because exophytic tumours are pre-potent. Survival rates are similar to those of patients with squamous cell carcinoma. PMID- 17410643 TI - Rapid genotyping of Toxoplasma gondii by pyrosequencing. AB - Most human infections with the protozoan parasite Toxoplasma gondii are asymptomatic, but severe symptoms can occur in immunocompromised patients, in developing foetuses, and in ocular infections in immunocompetent individuals. The majority of T. gondii strains can be divided into three main lineages, denoted types I, II and III, which are known to cause different clinical presentations. Simple molecular methods with the capacity to discriminate rapidly among strains may help to predict the course of infection and influence the choice of treatment. In the present study, real-time PCR followed by pyrosequencing was used to discriminate among types I, II and III by analysis of two single nucleotide polymorphisms in the GRA6 gene. Twenty-one isolates of T. gondii characterised previously were analysed. Three different GRA6 alleles detected by the pyrosequencing technique identified types I, II and III isolates correctly, while four atypical isolates possessed either the GRA6 allele 1 or the GRA6 allele 3. Reproducibility was 100%, and typeability, when including atypical strains, was 81%. It was also possible to discriminate a mixture of two genotypes. The method was used to identify GRA6 type II in blood and lung tissue from an allogeneic transplant recipient with toxoplasmosis. PMID- 17410644 TI - Blood samples drawn for culture as a surrogate marker for case-mix adjustment of hospital antibiotic use. AB - Hospital antibiotic consumption is generally adjusted to occupancy. This study hypothesised that the number of blood culture samples could be a surrogate marker for case-mix adjustment. Antibiotic consumption was compared over 16 consecutive trimesters in one medical ward in terms of patient-days or blood culture samples. Compared with patient-days, measurement adjusted to blood culture samples detected three trimesters with an unusually high consumption, and one trimester with consumption falsely classified as high because of a high incidence of infections. Blood culture numbers enabled easy and accurate identification of periods with a drift in antibiotic consumption ina medical ward. PMID- 17410645 TI - Caspase activation and extracellular signal-regulated kinase/Akt inhibition were involved in luteolin-induced apoptosis in Lewis lung carcinoma cells. AB - Luteolin was isolated from Scutellaria barbata D. Don (S. barbata). In the present study, we examined the underlying molecular mechanism of luteolin and its effect on in vivo tumor growth of Lewis lung carcinoma (LLC) cells. Luteolin exhibited antiproliferative activity against LLC cells with IC50 of 12 microM. Luteolin effectively increased Annexin-V-positive cells as well as sub G1 DNA portion as seen on flow cytometric analysis. Western blotting has revealed that luteolin effectively activates caspase 9 and 3, cleaves poly (ADP-ribose) polymerase (PARP), and increases the ratio of Bax/Bcl-2. Furthermore, mitochondrial membrane potential was reduced by luteolin as seen on fluorescence microscopy. Luteolin downregulated the expression of extracellular signal regulated kinase (ERK) and Akt in a concentration-dependent manner. In addition, luteolin significantly inhibited the growth of LLC cells implanted on the flank of mice to 40% and 60% of untreated control group values at 2 mg/kg and 10 mg/kg, respectively. Similarly, luteolin significantly reduced the expression of proliferating cell nuclear antigen (PCNA) as well as increased the expression of terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) in tumor section of LLC-bearing mice as determined by immunohistochemistry. Taken together, these results suggest that luteolin exerts antitumor activity by caspase activation and ERK/Akt inhibition. PMID- 17410646 TI - Images in Congenital Heart Disease. Symptomatic divided right atrium in a newborn. PMID- 17410647 TI - Images in Congenital Heart disease. Right arch and bilateral arterial ducts in a patient with discordant atrioventricular connections with pulmonary atresia and non-confluent pulmonary arteries. PMID- 17410648 TI - Images in Congenital Heart Disease. Infective endocarditis of the patent oval fossa assessed by three-dimensional echocardiography. PMID- 17410649 TI - Antiplatelet activity of carnosic acid, a phenolic diterpene from Rosmarinus officinalis. AB - Carnosic acid is a major phenolic diterpene derived from Rosmarinus officinalis and has been reported to have antioxidant, antibacterial, anticancer, antiobese and photoprotective activities. This study investigated the antiplatelet activity of carnosic acid. carnosic acid significantly inhibited collagen-, arachidonic acid-, U46619- and thrombin-induced washed rabbit platelet aggregation in a concentration-dependent manner, with IC50 values of 39+/-0.3, 34+/-1.8, 29+/-0.8 and 48+/-2.9 microM, respectively, while it failed to inhibit PMA-(a direct PKC activator) and ADP-induced platelet aggregation. In agreement with its antiplatelet activity, carnosic acid blocked collagen-, arachidonic acid-, U46619 and thrombin-mediated cytosolic calcium mobilization. accordingly, serotonin secretion and arachidonic acid liberation were also inhibited in a similar concentration-dependent manner. However, in contrast to the inhibition of arachidonic acid-induced platelet aggregation, carnosic acid had no effect on the formation of arachidonic acid-mediated thromboxane A2 and prostaglandin D2, thus indicating that carnosic acid has no effect on the cyclooxygenase and thromboxane A2 synthase activity. Overall, these results suggest that the antiplatelet activity of carnosic acid is mediated by the inhibition of cytosolic calcium mobilization and that carnosic acid has the potential of being developed as a novel antiplatelet agent. PMID- 17410650 TI - Staying power. PMID- 17410651 TI - [Pulmonary hypertension: tailored treatment]. PMID- 17410652 TI - No more coding! PMID- 17410653 TI - Family medicine and innovation. PMID- 17410654 TI - Vital concerns. Bringing up depression with your doctor. PMID- 17410655 TI - Supplements. Hope or hype? PMID- 17410657 TI - Healthy eating. Feeling bored? Add pizzazz to your pantry. PMID- 17410656 TI - Aerobic exercise. What it is and why it's good. PMID- 17410658 TI - Trailblazers. Recent years have seen the addition of several new classes of diabetes drugs, each one heralded as "The Next Big Thing" in diabetes. Have they lived up to their promise? PMID- 17410659 TI - Set a goal to take control. Aim for lower blood glucose levels to feel better now and stay healthy for years to come. PMID- 17410660 TI - Preventing complications. Preventive care and early awareness yield a healthier, less "complicated" life. PMID- 17410661 TI - A good heart. What it is and how to get it. PMID- 17410662 TI - What's up doc? How to make the most of your health care visit. PMID- 17410663 TI - Finger-lickin' chicken. Here are some clever ideas for jazzing up a classic food. PMID- 17410664 TI - Research profile. Candid conversations. Patients and physicians collaborate. Victor Montori, MD. PMID- 17410665 TI - Nutrition for dental health. PMID- 17410666 TI - Continuous glucose monitoring--getting started. PMID- 17410667 TI - Early spring harvest recipes. PMID- 17410668 TI - How to avoid errors in diabetes care. PMID- 17410669 TI - Mindful eating. Tuning in to your food. PMID- 17410670 TI - Diabetes quiz. How much do you know about obstructive sleep apnea? PMID- 17410671 TI - Increasing insulin sensitivity. PMID- 17410672 TI - Going with the (whole) grain. PMID- 17410673 TI - Keeping feet healthy. PMID- 17410674 TI - Supermarket smarts. Nondairy milks. PMID- 17410675 TI - Managing diabetes while breast-feeding. PMID- 17410676 TI - Retraction: Li Qiurong, Ma Jian, Tan Li, et al. Effect of docosahexaenoic acid on interleukin-2 receptor signaling pathway in lipid rafts, Sci China Ser C-Life Sci, 2006, 49(1): 63-72. PMID- 17410677 TI - [Treatment of acute otitis media in paediatrics: a meta-analysis]. AB - Otitis represents the second most common infection of the upper respiratory tract, its treatment being the most common cause for prescribing antibiotics in the United States. A large number of antimicrobials, especially beta-lactams and macrolides, are generally used for treating acute otitis media (AOM) in paediatric patients, owing to their antibacterial spectrum including the main aetiological pathogens. Efficacy, safety and compliance of Cefaclor were compared with those of other antibiotics in the treatment of paediatric AOM in a meta analysis of randomized controlled trials published between 1981 and 2004. Overall, evaluations were performed on 24 studies (Medline/PubMed, keywords "Cefaclor and otitis") which proved eligible (jadad score > or = 1); sixteen out of the 24 studies were multicentre, seven were double-blind. Mostly, the comparator agent was a beta-lactam, in four and three cases it was a macrolide or the association trimethoprim-sulfamethoxazole, respectively. Efficacy and safety were end-points of all studies whereas only 9 studies evaluated compliance. For the majority of studies (16/24) Cefaclor was administered for 10-day course. The analysis was based on a 2 x 2 contingency table with classification by treatment and number of improvements/cures, side-effects, and compliance of individual studies. The global estimate of the effective treatments was obtained with the weighted mean of the log OR (Odd Ratio) according to Mantel-Haenszel and associated confidence intervals (CI) at 95%. All the calculations were performed using SAS v.8. Chi-square test was performed. Clinical efficacy evaluation, number of improvements/cures, did not evidence a statistically significant difference among Cefaclor and comparators (86.8% vs 88.7%; Odds Ratio 0.77, IC 0.61/0.94). In the Cefaclor-treated patients, adverse events were observed in a statistically significant lower percentage compared to other antibiotics: 13.3% vs 19.4% (P < 0.0001), diarrhoea and gastro-intestinal disturbances being the most frequently observed. Compliance was observed in a similar proportion in the two patient groups, Cefaclor and comparators (Cefaclor 88.1; comparators 91.1%) and the slight difference was not statistically significant (Odds Ratio 0.77, IC 0.39-1.15). The present meta-analysis proves that in the treatment of paediatric AOM Cefaclor exhibits a clinical efficacy equal to other antibiotics usually employed in this setting, similar compliance but superior safety. PMID- 17410678 TI - Regulatory beneficiaries and informal agency policymaking. AB - Administrative agencies frequently use guidance documents to set policy broadly and prospectively in areas ranging from Department of Education Title IX enforcement to Food and Drug Administration regulation of direct-to- consumer pharmaceutical advertising. In form, these guidances often closely resemble the policies agencies issue in ordinary notice-and-comment rulemaking. However, guidances are generally developed with little public participation and are often immune from judicial review. Nonetheless, guidances can prompt significant changes in behavior from those the agencies regulate. A number of commentators have guardedly defended the current state of affairs. Though guidances lack some important procedural safeguards, they can help agencies supervise low-level employees and supply valuable information to regulated entities regarding how an agency will implement a program. Thus far, however, the debate has largely ignored the distinct and substantial interests of regulatory beneficiaries--those who expect to benefit from government regulation of others. Regulatory beneficiaries include, among others, pharmaceutical consumers, environmental users, and workers seeking safe workplaces. When agencies make policy informally, regulatory beneficiaries suffer distinctive losses to their ability to participate in the agency's decision and to invoke judicial review. This Article argues that considering the interests of regulatory beneficiaries strengthens the case for procedural reform. The Article then assesses some possible solutions. PMID- 17410679 TI - A new test for evaluating Eighth Amendment challenges to lethal injections. AB - An explosion of Eighth Amendment challenges to lethal injection protocols has struck the federal courts. The Supreme Court's recent decision in Hill v. McDonough,1 which empowered prisoners to bring challenges to lethal injection procedures under 42 U.S.C. para. 1983, has facilitated a flood of new lethal injection cases. In response, several courts have ordered states to alter their protocols, spurring other capital inmates to litigate such challenges. Distressingly, the courts evaluating these claims have almost no law to guide them. The last Supreme Court decision applying the Eighth Amendment to a method of execution was written in 1947; that case, Louisiana ex rel. Francis v. Resweber,2 occurred before the Eighth Amendment was applied to the states and resulted in a 4-1-4 split. Although lower courts have heard numerous challenges to execution methods, few have analyzed the constitutional validity of a method of execution in detail. Making matters worse, courts that find Eighth Amendment violations must craft equitable remedies that often amount to entirely new execution protocols. No clear precedent exists to guide courts in formulating such remedies. This Note proposes a legal standard for the administration of Eighth Amendment method-of-execution claims, focusing on lethal injection cases. Part I describes lethal injection procedures and summarizes recent litigation. Part II discusses the difficulty of evaluating lethal injection claims, analyzing both general difficulties in interpreting the Eighth Amendment and specific difficulties associated with lethal injection cases. Part III proposes a standard for addressing method-of-execution claims that attempts to balance a prisoner's interest in a painless execution with a state's interest in conducting executions efficiently. Part IV discusses remedies for unconstitutional procedures. Part V concludes. PMID- 17410680 TI - BVD virus causes heavy losses in a Scottish cattle herd. PMID- 17410681 TI - Reforming the food safety system: what if consolidation isn't enough? PMID- 17410682 TI - Teens living in disadvantaged neighborhoods lack access to parks and get less physical activity. PMID- 17410683 TI - CMS threatens termination. Patient death spurs inquiry of doc-owned hospital. PMID- 17410684 TI - Monosynaptic inhibition of the intracerebellar nuclei induced from the cerebellar cortex. PMID- 17410685 TI - Commentary on "Ito M, Yoshida M, Obata K. Monosynaptic inhibition of the intracerebellar nuclei induced from the cerebellar cortex. Experentia. 1964:20;575-6". PMID- 17410688 TI - Readers' responses to "is national health insurance coming to the United States?". PMID- 17410687 TI - Accelerating progress on salt iodisation in Sudan: time for action. AB - This paper uses a public health approach to examine briefly: (a) the progress of universal salt iodisation (USI) in Sudan; (b) the roles of the main actors involved; and (c) the main issues around accelerating USI. The literature, especially that coming from the UN agencies, is analysed and experiences from the recently revitalised USI programme, and related relevant meetings, are distilled. In Sudan the prevalence of goitre is 22 per cent. It is assumed that productivity among the people affected is reduced by 5-25 per cent. Little apparent progress has been made with USI. The Government of Sudan, UN multilateral agencies, international consultative groups, bilateral agencies, global and national non governmental organisations and, increasingly, the private sector must work together to find innovative approaches to increase awareness of the broader social, public health and nutritional contexts, and to advocate for increased national nd international funding. PMID- 17410686 TI - Should symptomatic menopausal women be offered hormone therapy? AB - Many physicians remain uncertain about prescribing hormone therapy for symptomatic women at the onset of menopause. The American Society for Reproductive Medicine (ASRM) convened a multidisciplinary group of healthcare providers to discuss the efficacy and risks of hormone therapy for symptomatic women, and to determine whether it would be appropriate to treat women at the onset of menopause who were complaining of menopausal symptoms. MAJOR FINDINGS: Numerous controlled clinical trials consistently demonstrate that hormone therapy, administered via oral, transdermal, or vaginal routes, is the most effective treatment for vasomotor symptoms. Topical vaginal formulations of hormone therapy should be preferred when prescribing solely for the treatment of symptoms of vulvar and vaginal atrophy. Data from the Women's Health Initiative indicate that the overall attributable risk of invasive breast cancer in women receiving estrogen plus progestin was 8 more cases per 10,000 women-years. No increased risk for invasive breast cancer was detected for women who never used hormone therapy in the past or for those receiving estrogen only. Hormone therapy is not effective for the treatment of cardiovascular disease and that the risk of cardiovascular disease with hormone therapy is principally in older women who are considerably postmenopause. CONCLUSIONS: Healthy symptomatic women should be offered the option of hormone therapy for menopausal symptoms. Symptom relief with hormone therapy for many younger women (at the onset of menopause) with menopausal symptoms outweighs the risks and may provide an overall improvement in quality of life. Hormone therapy should be individualized for symptomatic women. This involves tailoring the regimen and dose to individual needs. PMID- 17410689 TI - Readers' responses and authors' reply to "medical marijuana: politics trumps science at the FDA". PMID- 17410690 TI - Readers' and author's responses to "ethics and anomalous (alternative) medical practices". PMID- 17410691 TI - Fatal emphysematous cholecystitis caused by clostridium perfringens. PMID- 17410692 TI - Family member involvement in hastened death. AB - When patients pursue a hastened death, how is the labor of family care-giving affected? The authors examined this question in a qualitative study of 35 families. Four cases reveal the main themes: "taking care" included mutual protection between patients and family members; "midwifing the death" without professional support left families unprepared for adverse events; "tying up loose ends" included dealing with family members' fear of legal consequences; and "moving ahead" involved a greater risk of complicated grief when families encountered complications during the dying process. These results highlight the positive and negative consequences of family members' participation in a hastened death. PMID- 17410693 TI - Hope theory: a framework for understanding suicidal action. AB - This article examines C. R. Snyder's (1994, 2000a) theory of hope and its application for understanding suicide. Strengths, weaknesses, and gaps in the suicide literature are outlined, and A. T. Beck's theory of hopelessness is compared with Snyder's hope theory. Hope theory constructs are used to examine the relationship of suicide to hope/hopelessness, goals, pathways thinking, and agency thinking. This critical review is intended to broaden our theoretical understanding of suicide and is meant to form the basis for future empirical investigation of suicide-related behavior using the framework of hope theory. Implications for suicide prevention programs and approaches to treating suicidal individuals are outlined. PMID- 17410694 TI - Examination of proposed criteria for complicated grief in people confronted with violent or non-violent loss. AB - In the late 1990s, a panel of experts proposed criteria for complicated grief (CG) and found these to have satisfactory operating characteristics. The present study aimed to replicate that finding in 4 groups of mourners divided by cause of loss and time from loss. Data were available from 1,052 bereaved individuals. All had completed the revised Inventory of Complicated Grief through the Internet. Receiver operator characteristic analysis was used to analyze data. Results showed that, in all 4 groups, the proposed criteria for CG performed well at distinguishing between people who did and did not meet criteria for caseness of CG. PMID- 17410695 TI - Life as death scholars: passion, personality, and professional perspectives. AB - In death-averse American society, the field of thanatology is often socially and academically isolating. The purpose of this article is to describe the experiences of a group of death scholars and share insights gained as members of an interdisciplinary team. They discuss the ways in which they have created a special "safe" space for death study collaboration. The melding of their individual passions, personalities, and professional perspectives has allowed them to form a productive and satisfying work environment. Additional discourse in this field is needed on the nature and processes of such death-studies groups. PMID- 17410696 TI - Biologic agents in rheumatology: safety considerations. PMID- 17410697 TI - Current treatment for psoriatic arthritis and other spondyloarthritides. PMID- 17410698 TI - Comparison of the efficacy of biologic therapy for rheumatoid arthritis: can the clinical trials be accurately compared? PMID- 17410699 TI - Quantitative measures to assess patients with rheumatic diseases: 2006 update. PMID- 17410700 TI - Pain control after craniotomy: off balance on the tightrope? PMID- 17410701 TI - Prospective evaluation of pain and analgesic use following major elective intracranial surgery. AB - OBJECT: Opioid administration after major intracranial surgery is often limited by a presumed lack of need and a concern that opioids will adversely affect the postoperative neurological examination. The authors conducted a prospective study to evaluate the incidence, severity, and treatment of postoperative pain in patients who underwent major intracranial surgery. METHODS: One hundred eighty seven patients (77 men and 110 women, mean age 52 +/- 15 years, mean weight 78.1 +/- 19.9 kg) underwent either supratentorial (129 patients) or infratentorial (58 patients) procedures. Sixty-nine percent of the patients reported experiencing moderate to severe pain (> or =4 on a 0-10 scale) during the 1st postoperative day. Pain scores greater than or equal to 4 persisted in 48% on the 2nd postoperative day. Approximately 80% of patients were treated with acetaminophen on the 1st postoperative day, whereas opioids (primarily intravenous fentanyl) were administered to 58%. Compared with patients who underwent supratentorial procedures, those who underwent infratentorial procedures reported more severe pain at rest (mean score 4.9 +/- 2.2 compared with 3.8 +/- 2.6; p = 0.015) and with movement (mean score 6.3 +/- 2.6 compared with 4.5 +/- 2.7; p < 0.001) on the 1st postoperative day. On both the 1st and 2nd postoperative days, patients who underwent infratentorial procedures received greater quantities of opioid (p < or = 0.019) and nonopioid (p < or = 0.013) analgesics than those who underwent supratentorial procedures. Patients' dissatisfaction with analgesic therapy was significantly associated with elevated pain levels on the first 2 postoperative days (p < 0.001). CONCLUSIONS: In contrast to prevailing assumptions, the study findings reveal that most patients undergoing elective major intracranial surgery will experience moderate to severe pain for the first 2 days after surgery and that this pain is often inadequately treated. PMID- 17410702 TI - Thirty-day morbidity and mortality rates for carotid artery intervention by surgeons who perform both carotid endarterectomy and carotid artery angioplasty and stent placement. AB - OBJECT: There is no known standard 30-day morbidity and mortality rate for high risk patients undergoing carotid artery (CA) angioplasty and stent (CAS) placement. The high-risk registries and the Stenting and Angioplasty with Protection in Patients at High Risk for Endarterectomy, Carotid Revascularization using Endarterectomy or Stenting Systems, and European Long-term Carotid Artery Stenting trials report different rates of morbidity and mortality, and each high risk cohort has a different risk profile. The applicability of carotid endarterectomy (CEA) results from North American Symptomatic Carotid Endarterectomy Trial/Asymptomatic Carotid Atherosclerosis Study (NASCET/ACAS) remains uncertain, as most clinical CAS placement series reported to date typically included patients who would not have qualified for those studies. At the University at Buffalo, the same neurosurgeons perform triage in patients with CA disease and perform both CEA and CAS insertion. The authors review morbidity and mortality rates in this practice model. METHODS: Diagnosis-related group codes were used to search the authors' practice database for patients who had undergone a completed CA intervention solely for the indication of atherosclerotic disease. One hundred twenty patients (129 vessels) treated with CAS surgery and 95 patients (100 vessels) treated with CEA met these criteria. In the CAS placement group, 78% of the patients would not have met NASCET/ACAS inclusion criteria. Demographic and clinical data for both groups were recorded on a spreadsheet for analysis. At 30 days, one patient in the CEA group and two in the CAS group had died. Stroke occurred in one patient in the CAS group and none in the CEA group. Myocardial infarction (MI) occurred in one patient who underwent CAS surgery compared with three undergoing CEA. Composite incidence of stroke/death/MI was 3.3% in the CAS group and 3.2% in the CEA group. CONCLUSIONS: In a practice in which surgeons perform both CEA and CAS surgery, the event rates for the CAS surgery equivalent to NASCET and ACAS rates for CEA can be achieved, even in high-risk NASCET/ACAS-ineligible patients in 78% of the CAS cases. PMID- 17410703 TI - Midterm outcomes of paclitaxel-eluting stents for the treatment of intracranial posterior circulation stenoses. AB - OBJECT: Symptomatic intracranial vertebral and basilar artery atherosclerotic stenoses carry a high risk of stroke and permanent disability if refractory to maximal medical therapy. The authors conducted a study to determine the technical feasibility and midterm clinical and angiographic outcomes in patients in whom paclitaxel-eluting stents were placed for the treatment of symptomatic intracranial posterior circulation stenoses. METHODS: A retrospective review of medical records and imaging studies was performed for 13 consecutive patients in whom paclitaxel-coated stents were used to treat symptomatic posterior circulation intracranial stenoses between 2002 and 2005. Clinical follow-up data were supplemented by telephone interviews. The technical success rate for stent placement was 100%. One patient (8%) suffered a periprocedural stroke. Twelve patients (92%) underwent clinical follow up for a minimum of 3 months postsurgery, and 11 (92%) of these patients remained asymptomatic after a mean period of 10.9 months. Nine patients (69%) underwent catheter angiographic follow up, and no patient had significant in-stent recurrence of stenosis after a mean period of 5.4 months. CONCLUSIONS: Treatment of intracranial posterior circulation stenoses with drug-eluting stents is technically feasible, and the rate of clinically significant periprocedural complications is low. Rates of stenosis recurrence are reduced compared with those of bare-metal stents in the midterm. Midterm clinical outcome is excellent; no symptom recurrence was observed in this patient cohort. PMID- 17410704 TI - A prospective single-center analysis of the safety and efficacy of the hydrocoil embolization system for the treatment of intracranial aneurysms. AB - OBJECT: The HydroCoil embolization system is a helical platinum coil coated with a polymeric hydrogel that expands when it contacts aqueous solutions to increase filling volumes, improve mesh stability, and possibly elicit a healing response within the aneurysm. In this paper, the authors report the 1-year recurrence and complication rates of 67 aneurysms embolized with the HydroCoil system. METHODS: Sixty-four consecutive patients (67 total aneurysms) with small (< or =7 mm), large (8-15 mm), very large (16-24 mm), and giant (> or =25 mm) aneurysms in the anterior and posterior intracranial circulations were treated with HydroCoils between March 2003 and September 2004. All aneurysms were embolized by the senior author (A.S.B) with HydroCoils alone or in combination with bare platinum coils, until either there was no further angiographic contrast filling of the aneurysm or the microcatheter was pushed out of the dome by the coil mass. Balloon assistance was used in three cases and combined Neuroform stent-coil embolization in eight other cases. To evaluate the safety and 1-year efficacy of the HydroCoil system, periprocedural complications were recorded, and angiographic recurrences were categorized using the Raymond-Roy Occlusion Classification (RROC) system. The 1-year aneurysm recurrence rate independent of size was 15% in patients treated with HydroCoils. Seventy percent of the patients had stable occlusions. The recurrence rate for small aneurysms was 3.7%, and the combined recurrence rate for small and large aneurysms was 6%. Fifteen percent of the aneurysms initially categorized as RROC Type 2 or 3 with stasis of contrast material at the time of initial embolization improved in RROC type, allowing the authors to develop the aneurysm embolization grade to predict recurrence. The neurological complication rate was 14.9%, of which 4.5% represented permanent neurological deficits. CONCLUSIONS: The HydroCoil embolization system is safe and provides excellent 1-year occlusion of small and large aneurysms with initial RROC Type 1, as well as those with RROC Types 2 and 3 with stasis of contrast material at the time of embolization. Very large and giant aneurysms were not as successfully occluded with this system. Treatment of large and giant internal carotid artery aneurysms was more likely to result in cranial nerve palsies and postembolization headaches than treatment in other locations. The aneurysm embolization grade the authors developed using the results of this study accurately predicted 1-year recurrence rates based on the immediate postembolization angiographic characteristics of the treated aneurysm. PMID- 17410705 TI - Trends in the incidence of subarachnoid hemorrhage in Akita Prefecture, Japan. AB - OBJECT: The incidence of subarachnoid hemorrhage (SAH) in Japan has been reported to be among the highest in the world. The aim of this study was to analyze the incidence of SAH and its trends in Akita Prefecture, Japan. METHODS: The incidence rate of SAH between January 1, 1989, and December 31, 1998, was investigated using the prospective Akita Stroke Register. Computed tomography scans were obtained in all suspected cases of SAH. During the study period, 3257 patients with their first SAH were registered. The crude annual incidence rates of SAH in men, women, and both sexes were 20, 33, and 27 per 100,000 population per year, respectively. Age-standardized incidence rates remained stable. Crude incidence rates in women increased. In men the incidence reached a plateau after 55 years of age; in women it peaked in the eighth decade of life. The 28-day case fatality rate was 26.7%, without significant changes. Two diurnal peaks were observed: one at 7 a.m. and another at 5 p.m. The incidence was lowest in summer and highest in spring, in both sexes. CONCLUSIONS: The incidence of SAH in patients in Akita Prefecture is similar to that reported in other Japanese studies. The age-standardized incidence rates are stable. The increasing crude incidence in women is probably attributable to the aging of the Japanese population. PMID- 17410706 TI - Optimal reverse trendelenburg position in patients undergoing craniotomy for cerebral tumors. AB - OBJECT: To the authors' knowledge, repeated measurements of intracranial pressure (ICP), cerebral perfusion pressure (CPP), and the degree of dural tension during different positions on the operating table (reverse Trendelenburg position [rTp]) have not been studied in patients undergoing craniotomy. METHODS: In the present study 53 patients with supratentorial cerebral tumors who underwent craniotomy in the supine position were included. Subdural ICP, mean arterial blood pressure (MABP), CPP, and jugular bulb (JB) pressure were recorded, and the degree of dural tension was analyzed while patients were in the neutral operating position and at 5, 10, and 15 degrees rTp. The optimal operating position was defined as the one at which subdural ICP was as low as possible, and CPP was greater than or equal to 60 mm Hg or as high as possible. Subdural ICP, MABP, and JB pressure decreased significantly after each 5 degrres change in rTp compared with the preceding position. Dural tension decreased significantly up to 10 degrees rTp, but was unchanged at 15 degrees rTp. At 5 degrees rTp CPP remained unchanged, but it decreased significantly during 10 and 15 degrees rTp. The optimal position in the majority of patients was determined to be 15 degrees rTp. CONCLUSIONS: Before opening the dura mater for craniotomy, repeated measurements of ICP and CPP, in the neutral position and at 5, 10, and 15 degrees rTp, provide valuable information regarding the optimal level of ICP and CPP. PMID- 17410707 TI - Lack of association between the apolipoprotein E gene and aneurysmal subarachnoid hemorrhage in an Italian population. AB - OBJECT: The results of genome-wide scan studies have suggested the presence of a genetic risk factor for aneurysmal subarachnoid hemorrhage (SAH) on chromosome 19 (at 19p13). The apolipoprotein E (APOE) gene is located in this chromosomal region and encodes a protein that exerts several neuroprotective and neurotrophic functions in the brain. The purpose of this study was to evaluate whether a particular allele or genotype of the APOE gene would modify the occurrence or the clinical features of SAH. METHODS: Genomic DNA was extracted from 146 patients with aneurysmal SAH and 222 age- and sex-matched healthy controls and genotyped for the triallelic polymorphism of the APOE gene (epsilon2, epsilon3, and epsilon4). Allele and genotype frequencies were compared between patients and controls. The clinical characteristics of the disease were compared according to the different APOE genotypes. Allele and genotype frequencies of the APOE gene polymorphism were nearly identical in cases and controls. Patients carrying the APOE epsilon4 allele had a significantly higher Hunt and Hess grade on admission (p = 0.0014). There was no significant relationship between any of the other clinical characteristics and the APOE genotype. CONCLUSIONS: The authors' data do not support the hypothesis that genetic variations within the APOE gene are associated with aneurysmal SAH. However, the APOE gene influences the disease phenotype and may be regarded as a disease modifier gene. PMID- 17410708 TI - Predictors of hematoma enlargement in patients with intracerebral hemorrhage treated with rapid administration of antifibrinolytic agents and strict blood pressure control. AB - OBJECT: Hematoma enlargement is a major cause of poor outcome in patients with intracerebral hemorrhage (ICH). A combination of rapid administration of antifibrinolytics and strict blood pressure (BP) control for prevention of hematoma enlargement has been recently reported. The authors examined the incidence and predictors of hematoma enlargement in patients with ICH who were treated with this therapy. METHODS: Rapid administration of antifibrinolytic agents consisted of intravenous administration of 2 g tranexamic acid over 10 minutes. Systolic BP was strictly maintained below 150 mm Hg using intravenous nicardipine. Immediately after diagnosis of ICH on computed tomography (CT), 188 patients who were admitted within 24 hours of symptom onset were treated with a combination of rapid administration of antifibrinolytic agents and BP control. Hematoma enlargement was determined on the basis of a second CT scan performed the day after admission. Several factors, including those that have been reported to affect hematoma enlargement, were compared between patients with and without hematoma enlargement. Hematoma enlargement (> or =20% volume increase) was observed in eight (4.3%) of 188 patients. Previous use of antiplatelet agents was significantly more frequent in patients with hematoma enlargement (p < 0.05). No significant between-group difference was found for any other factors Conclusions. Previous use of antiplatelet agents was a predictor of hematoma enlargement in patients with ICH treated with rapid administration of antifibrinolytic agents and BP control. PMID- 17410709 TI - Emergency embolectomy for treatment of acute middle cerebral artery occlusion. AB - OBJECT: The authors evaluated the efficacy of emergency embolectomy in patients with acute middle cerebral artery occlusion (MCAO). METHODS: A retrospective review of the charts for patients undergoing embolectomy at the authors' institution was performed. Between October 1997 and May 2004, 12 patients (mean age 70 years) with acute MCAO were treated using embolectomy. Local intraarterial fibrinolysis with urokinase was initially undertaken in eight of 12 patients. Sufficient recanalization was not achieved with fibrinolysis in any patient, and thus embolectomy was performed immediately thereafter. Recanalization by embolectomy was achieved in all patients (mean occlusion time 6 hours, 11 minutes). Each patient's condition was evaluated on discharge. Outcomes in the 12 patients according to the Glasgow Outcome Scale were good recovery in five, moderate disability in two, severe disability in three, vegetative state in one, and death due to a cardiac complication in one. None of the 12 patients had symptomatic hemorrhagic infarction. CONCLUSIONS: Emergency embolectomy can be performed in patients with MCAO with minimal morbidity and death. The procedure can be used to achieve good recovery even in patients in whom fibrinolysis is insufficient after acute MCAO and should be a part of the algorithm for the treatment of MCAO. PMID- 17410710 TI - Ischemic complications associated with resection of opercular glioma. AB - OBJECT: Opercular glioma inferolateral to the hand/digit sensorimotor area can be resected safely using a neuronavigation system and functional brain mapping techniques. However, the surgery can still sometimes cause postoperative ischemic complications, the character of which remains unclear. The authors of this study investigated the occurrence of infarction associated with resection of opercular glioma and the arterial supply to this region. METHODS: The study involved 11 consecutive patients with gliomas located in the opercular region around the orofacial primary motor and somatosensory cortices but not involving either the hand/digit area or the insula, who had been treated in their department after 1997. Both pre- and postoperative diffusion-weighted magnetic resonance (MR) imaging was performed in the nine consecutive patients after 1998 to detect ischemic complications. All patients underwent open surgery for maximum tumor resection. Postoperative MR imaging identified infarction beneath the resection cavity in all patients. Permanent motor deficits associated with infarction involving the descending motor pathway developed in two patients. Cadaveric angiography showed that the distributing arteries to the corona radiata were the long insular arteries and/or medullary arteries from the opercular and cortical segments of the middle cerebral artery. CONCLUSIONS: Subcortical resection around the upper limiting sulcus of the posterior region of the insula and wide resection in the anteroposterior and cephalocaudal directions of the opercular region were considered to be risk factors of the critical infarction. Surgeons should be aware that resection of opercular glioma can disrupt the blood supply of the corona radiata, and carries the risk of permanent motor deficits. PMID- 17410711 TI - Intradural cranial chordoma: a rare presentation of an uncommon tumor. Surgical experience and review of the literature. AB - OBJECT: Intracranial intradural chordomas are extremely rare lesions, and only a few cases have been reported in the neurosurgical literature. The authors performed a retrospective analysis of cases treated at their institutions. They present an illustrative surgical scenario and discuss the published literature, pathogenesis, and histopathological features as well as available follow-up data on the clinical behavior of these intradural lesions. METHODS: The authors reviewed clinical, neuroimaging, operative and follow-up data from a series of 79 chordomas treated at their institutions over the last 17 years. They found that the tumors were confined exclusively to the intradural compartment in only three cases. Staining for MIB-1 was performed to support the differential diagnosis between ecchordosis physaliphora and intradural chordoma, and in all three patients the neoplastic nature of the lesions was confirmed. None of these three cases showed recurrence of the lesion at midterm follow up. CONCLUSIONS: Complete resection, followed by close clinical and neuroradiological follow up, is warranted in cases of intradural cranial chordoma. PMID- 17410712 TI - Neuronal expression of Fas-associated death domain protein and caspase-8 in the perinidal parenchyma of cerebral arteriovenous malformations. AB - OBJECT: The expression and localization of phosphorylated Fas-associated death domain protein (pFADD) and cleaved caspase-8 was examined in human cerebral arteriovenous malformations (AVMs). The authors focused on the perinidal parenchyma to clarify the effect of AVMs on perinidal brain tissue. METHODS: Seventeen cerebral AVMs were analyzed using immunohistochemical methods. Specimens were removed from patients during surgical procedures. The characteristics of the areas that stained positively for pFADD or cleaved caspase 8 were also assessed using an image analysis system. Eleven (65%) of the 17 lesions demonstrated anti-pFADD immunoreactivity and 12 (71%) showed anti-cleaved caspase-8 immunoreactivity. The immunoreactive cells in the perinidal parenchyma demonstrated obvious neuronal morphological characteristics. The characteristics of pFADD-positive and cleaved caspase-8-positive areas were assessed using the image analysis system. The mean distance from the nidus adjacent to either area was not affected by preoperative hemorrhage. The neuronal densities of pFADD positive and cleaved caspase-8-positive areas were analyzed using the same system. The density of the control area (samples that were pFADD-negative and cleaved caspase-8 negative) was significantly higher when compared with that of pFADD-positive and cleaved caspase-8-positive areas (p < 0.05). The expressions of cleaved caspase-9, cleaved poly(adenosine diphosphate-ribose) polymerase, and apoptotic cells were analyzed using the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick-end labeling method. CONCLUSIONS: Neuronal areas that stained positively for pFADD and cleaved caspase-8 existed around the nidus of AVMs. In these areas, the neuronal density was lower than that in the other parenchyma around the AVM. Neuronal loss around the nidus may be the origin of brain dysfunction around AVMs. PMID- 17410713 TI - Detection of cerebral ischemia in neurovascular surgery using quantitative frequency-domain near-infrared spectroscopy. AB - OBJECT: There is great value in monitoring for signs of ischemia during neurovascular procedures. Current intraoperative monitoring techniques provide real-time feedback with limited accuracy. Quantitative frequency-domain near infrared spectroscopy (Q-NIRS) allows measurement of tissue oxyhemoglobin (HbO2), deoxyhemoglobin (HHb), and total hemoglobin (tHb) concentrations and brain tissue oxygen saturation (SO2), which could be useful when monitoring for evidence of intraoperative ischemia. METHODS: Using Q-NIRS, the authors monitored 25 neurovascular procedures including aneurysm clip placement, arteriovenous malformation resection, carotid endarterectomy, superficial temporal artery middle cerebral artery (MCA) bypass surgery, external carotid artery-MCA bypass surgery, encephaloduromyosynangiosis, and balloon occlusion testing. The Q-NIRS technology provides measurable cerebral oxygenation values independent from those of the scalp tissue. Thus, alterations in the variables measured with Q-NIRS quantitatively reflect cerebral tissue perfusion. Bilateral monitoring was performed in all cases. Five of the patients exhibited evidence of clinical ischemic events during the procedures. One patient suffered blood loss with systemic hypotension and developed diffuse brain edema intraoperatively, one patient suffered an ischemic event intraoperatively and developed an occipital stroke postoperatively, and one patient showed slowing on electroencephalography intraoperatively during carotid clamping; in two patients balloon occlusion testing failed. In all cases of ischemic events occurring during the procedure, Q NIRS monitoring showed a decrease in HbO2, tHb, and SO2, and an increase in HHb. CONCLUSIONS: . Quantitative frequency-domain near-infrared spectroscopy provides quantifiable and continuous real-time information about brain oxygenation and hemodynamics in a noninvasive manner. This continuous intraoperative oxygenation monitoring is a promising method for detecting ischemic events during neurovascular procedures. PMID- 17410714 TI - Evaluation of extracranial-intracranial bypass using quantitative magnetic resonance angiography. AB - OBJECT: To date, angiography has been the primary modality for assessing graft patency following extracranial-intracranial bypass. The utility of a noninvasive and quantitative method of assessing bypass function postoperatively was evaluated using quantitative magnetic resonance (MR) angiography. METHODS: One hundred one cases of bypass surgery performed over a 5.5-year period at a single institution were reviewed. In 62 cases, both angiographic and quantitative MR angiographic data were available. Intraoperative flow measurements were available in 13 cases in which quantitative MR angiography was performed during the early postoperative period (within 48 hours after surgery). There was excellent correlation between quantitative MR angiographic flow and angiographic findings over the mean 10 months of imaging follow up. Occluded bypasses were consistently absent on quantitative MR angiograms (four cases). The flow rates were significantly lower in those bypasses that became stenotic or reduced in diameter as demonstrated by follow-up angiography (nine cases) than in those bypasses that remained fully patent (mean +/- standard error of the mean, 37 +/- 13 ml/minute compared with 105 +/- 7 ml/minute, p = 0.001). Flows were appreciably lower in poorly functioning bypasses for both vein and in situ arterial grafts. All angiographically poor bypasses (nine cases) were identifiable by absolute flows of less than 20 ml/minute or a reduction in flow greater than 30% within 3 months. Good correlation was seen between intraoperative flow measurements and early postoperative quantitative MR angiographic flow measurements (13 cases, Pearson correlation coefficient = 0.70, p = 0.02). CONCLUSIONS: Bypass grafts can be assessed in a noninvasive fashion by using quantitative MR angiography. This imaging modality provides not only information regarding patency as shown by conventional angiography, but also a quantitative assessment of bypass function. In this study, a low or rapidly decreasing flow was indicative of a shrunken or stenotic graft. Quantitative MR angiography may provide an alternative to standard angiography for serial follow up of bypass grafts. PMID- 17410715 TI - Prolonged survival of mice with established intracerebral glioma receiving combined treatment with peroxisome proliferator-activated receptor-gamma thiazolidinedione agonists and interleukin-2-secreting syngeneic/allogeneic fibroblasts. AB - OBJECT: In this study the authors explored the benefits of treating C57B1/6 mice with an established intracerebral glioma by combining immunotherapy with interleukin (IL)-2-secreting syngeneic/allogeneic fibroblasts administered into the tumor bed along with the chemotherapeutic agent pioglitazone, a thiazolidinedione (TZD). The TZDs are agonists of the peroxisome proliferator activated receptor-gamma. They have been found to exert antiproliferative effects on several transformed cell lines. Data from prior studies by these authors have revealed the immunotherapeutic properties of the IL-2-secreting fibroblasts in treating intracerebral gliomas in mice. METHODS: The sensitivity of GL261 glioma cells and primary astrocytes to pioglitazone was determined in vitro by incubating the cells with increasing amounts of the drug. Viability was assessed by measuring lactate dehydrogenase release, and effects on metabolism were determined by measuring superoxide production and levels of superoxide dismutase. The GL261 cells were injected intracerebrally into C57B1/6 mice, followed by treatment with pioglitazone either orally or intracerebrally into the tumor bed. The effect of the combined therapy was determined by injecting C57B1/6 mice with an established intracerebral GL261 glioma with IL-2-secreting allogeneic fibroblasts and pioglitazone directly into the tumor bed through a unique cannula system. Pioglitazone was found to induce cell death in GL261 glioma cells grown in vitro while causing only modest damage to astrocytes. The application of pioglitazone also resulted in a significantly greater induction of cellular superoxide in glioma cells than in astrocytes, which can activate apoptotic pathways. Pioglitazone administered intracerebrally (p < 0.05) but not orally was found to prolong survival in mice harboring an intracerebral glioma. Synergistic effects of combination therapy on prolonging survival were found in mice receiving both pioglitazone and IL-2-secreting fibroblasts (p < 0.005, compared with untreated animals). Pioglitazone induces metabolic and oxidative stresses that are tolerated by astrocytes but not glioma cells, which could account for selective vulnerability and increased sensitivity to IL-2, suggesting potential for the use of this Food and Drug Administration-approved drug in the treatment of brain tumors. CONCLUSIONS: The data indicate the beneficial effects of combination therapy using pioglitazone and immunotherapy in mice harboring intracerebral glioma. PMID- 17410716 TI - Magnetic resonance imaging of in vitro glioma cell invasion. AB - OBJECT: An understanding of single glioma cell invasion has been limited by the static picture provided by histological studies. The ability to nondestructively assess cell invasion dynamically in a full 3D volume would improve the quality and quantity of information available from both in vivo and in vitro experiments. The purpose of this study was to observe glioma cell invasion in a 3D in vitro model using a microimaging protocol at 1.5 tesla and to assess the uptake of micron-sized particles of iron oxide (MPIO) and the consequent effects on cell function. METHODS: Rat C6 glioma cells were labeled with MPIO to a sufficient extent to allow single cell detection in vitro without significant effects on cell proliferation or plating efficiency. When placed on agar-coated plates, the cells formed stable multicellular tumor spheroids (MCTSs), which were embedded in collagen type I gel and serially visualized using magnetic resonance (MR) imaging and phase-contrast microscopy over 8 days. The MCTSs initially appeared as large susceptibility artifacts on MR images, but within 2 days, as cells moved away from the main MCTS, small discrete areas of signal loss, possibly due to single cells, could be observed and tracked. CONCLUSIONS: Glioma cell invasion can be nondestructively observed using MR imaging. The sensitivity of MR imaging, along with its ability to represent full 3D volumes noninvasively over time, makes it ideal for longitudinal in vivo cell tracking studies. PMID- 17410717 TI - Addition of intravenous N-methyl-D-aspartate receptor antagonists to local fibrinolytic therapy for the optimal treatment of experimental intracerebral hemorrhages. AB - OBJECT: Fibrinolytic therapy with recombinant tissue plasminogen activator (rtPA) is considered a treatment option in patients with deep-seated intracerebral hemorrhage (ICH). Nevertheless, the results of animal experiments have shown that tPA exerts pleiotropic actions in the brain, including regulation of vasoactivity, amplification of calcium conductance by cleavage of the N-methyl-D aspartate (NMDA) receptor subunit, and activation of metalloproteinases, which increase excitotoxicity, damage the blood-brain barrier, and worsen edema. The authors investigated whether the noncompetitive NMDA receptor antagonist MK801 can be used as an adjuvant therapy in combination with rtPA to attenuate the unfavorable delayed edema formation and inflammation observed following rtPA therapy in an experimental porcine model of ICH. METHODS: Twenty pigs were used in this study; MK801 (0.3 mg/kg) was administered to each pig intravenously immediately after hematoma induction and on the 1st and 3rd day after hematoma induction. Ten of the 20 pigs were randomly assigned to fibrinolytic therapy with rtPA (MK801-tPA group), whereas in the remaining 10 control animals (MK801 group) the hematomas were allowed to follow their natural courses of resorption. The extent of edema formation was evaluated using magnetic resonance (MR) imaging volumetry on Days 0, 4, and 10 after hematoma induction and was compared with histopathological changes found at necropsy. The mean edema volumes in these two groups were also compared with that in the group of nine pigs examined in a preceding experimental series, in which the animals' hematomas were only treated with rtPA (tPA group). In the 10 animals in the MK801-tPA group, the mean perihematoma edema volume on MR images had not significantly increased by Day 4 (p < 0.08) or Day 10 (p < 0.35) after hematoma induction. In the 10 animals in the MK801 group, the increase in mean perifocal edema size was significant after 4 days (p < 0.001) and nonsignificant after 10 days (p < 0.09). In the nine animals in the tPA group, the mean edema volume significantly increased by Days 4 (p < 0.002) and 10 (p < 0.03). CONCLUSIONS: As suggested by the reduction in delayed edema volume and the inflammatory response, MK801 modifies the neurotoxic properties of rtPA but not those of blood degradation products. Possibly, fibrinolytic therapy of ICH is more beneficial if combined with agents such as MK801. PMID- 17410718 TI - Acute cerebral vascular injury after subarachnoid hemorrhage and its prevention by administration of a nitric oxide donor. AB - OBJECT: Structural changes in brain parenchymal vessels occur within minutes after subarachnoid hemorrhage (SAH). These changes include platelet aggregation, activation of vascular collagenases, and destruction of perivascular collagen IV. Because collagen IV is an important component of the basal lamina, the authors attempted to further define changes in vascular structure (length and luminal diameter) and their relationship to vascular permeability immediately after SAH. In addition, the authors explored whether such alterations were attenuated by administration of a nitric oxide (NO) donor. METHODS: Endovascular perforation was used to induce SAH in rats. Two sets of experiments were performed. The first established changes in vascular structure and permeability (collagen IV and endothelial barrier antigen [EBA] dual immunofluorescence) during the first 24 hours after SAH. In the second, the investigators examined the effects of an NO donor on vascular structure, permeability, and collagenase activity (in situ zymography). In this second study, animals received intravenous infusion of the NO donor S-nitrosoglutathione (GSNO, 1 microM/8 microl/min) 15 minutes after induction of SAH and were killed 3 hours after SAH onset. Controls were naive unoperated animals for the first study and saline-infused SAH animals for the second. The authors found a time-dependent decrease in area fraction, length, and luminal diameter of collagen IV- and EBA-immunofluorescent vessels after SAH. The greatest change occurred at 3 hours after onset of SAH. Administration of GSNO was associated with striking preservation of collagen IV and EBA immunofluorescence compared with saline treatment. Zymography indicated decreased collagenase activity in GSNO-treated SAH animals compared with saline-treated SAH animals. CONCLUSIONS: These results demonstrate changes in the structure and permeability of brain parenchymal microvessels after SAH and their reversal by treatment with an NO donor. PMID- 17410719 TI - Cerebral aneurysm progression suppressed by blockage of endothelin B receptor. AB - OBJECT: Cerebral aneurysm is a major cause of subarachnoid hemorrhage, but the mechanisms of its development remain unclear. Mechanical stretch has been reported to induce vascular smooth-muscle cell apoptosis via endothelin B receptors (ETBRs). The objectives of this study were to clarify the expression and localization of ETBR in cerebral aneurysms and to examine the effect of ETBR blockage on the development of experimental cerebral aneurysms. METHODS: Seventy two rats underwent a cerebral aneurysm induction procedure and were divided into four groups according to the duration of postoperative study periods. Expression of ETBR was confirmed by reverse transcription-polymerase chain reaction and immunohistochemical analysis. The authors also studied the effect of K-8794, an oral selective antagonist of ETBR, to see whether it would influence the formation of cerebral aneurysms. Two weeks after the aneurysm induction procedure, ETBR was rarely detected in anterior cerebral artery-olfactory artery bifurcations, but it was weakly expressed in experimental cerebral aneurysms at 1 month after the procedure, and markedly expressed at 3 months. The administration of K-8794 for 1 month after the procedure significantly reduced the number of advanced aneurysms and the number of apoptotic smooth-muscle cells. CONCLUSIONS: These results suggest that ETBR might play a significant role in the progression of cerebral aneurysms and have the potential to improve prevention and treatment of cerebral aneurysms. PMID- 17410720 TI - Erythropoietin and astrocytomas. PMID- 17410721 TI - Survival and invasiveness of astrocytomas promoted by erythropoietin. AB - OBJECT: The hypoxia-inducible pleiotropic hormone, erythropoietin (EPO), has recently been found to promote the development and survival of neurons and astrocytes. Since hypoxia has been implicated in the malignant progression of some human cancers, the authors investigated whether EPO signaling influenced the malignant properties of human astrocytoma cells. METHODS: Reverse transcriptase polymerase chain reaction, Western blot analysis, and immunohistochemical studies were used to measure EPO and its receptor (EPOR). Cell viability, Matrigel invasion assays, metalloprotease assays, EPO neutralizing antibodies, and EPOR overexpression were used to study the biological actions of EPO. Expression of both EPO and EPOR was observed in the hypoxic regions and invasive margins of glioma specimens obtained at biopsy, and expression of EPOR correlated with the stage of the tumor. The EPOR was also functionally upregulated by hypoxia in cultured glioblastoma multiforme (GBM) cells. Both hypoxia and EPO protected cultured GBM cells from cisplatin cytotoxicity and promoted the invasiveness of GBM cells through Matrigel by potentiating metalloprotease activity. Hypoxia enhanced cell invasion was attenuated in cells that overexpressed a nonfunctional EPOR. CONCLUSIONS: Hypoxia-inducible autocrine and paracrine EPO signaling participates in the malignant progression of GBMs. PMID- 17410722 TI - Image-guided convection-enhanced delivery of gemcitabine to the brainstem. AB - OBJECT: To determine if the potent antiglioma chemotherapeutic agent gemcitabine could be delivered to the brainstem safely at therapeutic doses while monitoring its distribution using a surrogate magnetic resonance (MR) imaging tracer, the authors used convection-enhanced delivery to perfuse the primate brainstem with gemcitabine and Gd-diethylenetriamine pentaacetic acid (DTPA). METHODS: Six primates underwent convective brainstem perfusion with gemcitabine (0.4 mg/ml; two animals), Gd-DTPA (5 mM; two animals), or a coinfusion of gemcitabine (0.4 mg/ml) and Gd-DTPA (5 mM; two animals), and were killed 28 days afterward. These primates were observed over time clinically (six animals), and with MR imaging (five animals), quantitative autoradiography (one animal), and histological analysis (all animals). In an additional primate, 3H-gemcitabine and Gd-DTPA were coinfused and the animal was killed immediately afterward. In the primates there was no histological evidence of infusate-related tissue toxicity. Magnetic resonance images obtained during infusate delivery demonstrated that the anatomical region infused with Gd-DTPA was clearly distinguishable from surrounding noninfused tissue. Quantitative autoradiography confirmed that Gd DTPA tracked the distribution of 3H-gemcitabine and closely approximated its volume of distribution (mean volume of distribution difference 13.5%). Conclusions. Gemcitabine can be delivered safely and effectively to the primate brainstem at therapeutic concentrations and at volumes that are higher than those considered clinically relevant. Moreover, MR imaging can be used to track the distribution of gemcitabine by adding Gd-DTPA to the infusate. This delivery paradigm should allow for direct therapeutic application of gemcitabine to brainstem gliomas while monitoring its distribution to ensure effective tumor coverage and to maximize safety. PMID- 17410723 TI - [Eugeniu Proca--founder of modern urological school and pioneer of transplantation]. PMID- 17410724 TI - [Disease-related malnutrition and its influence on perioperative outcome]. AB - Disease-related malnutrition is highly prevalent among surgical patients and has important deleterious effects on postoperative outcome: increased complications rate, length of hospital stay, mortality and costs. Malnutrition (equivalent with undernutrition in the surgical patients) can be present already on admission, could rapidly develop or aggravate during hospital stay. The determination of malnutrition in this high-risk group of patients in multifactorial: decreased intake (anorexia, gastrointestinal symptoms, prolonged perioperative fasting) and increased demands (hyper-catabolism due to underlying disease, surgical stress, eventual postoperative complications). The identification of patients at nutritional risk by screening on admission and whenever the patient situation changes throughout hospitalization is the first, mandatory step for developing an accurate nutritional care plan in these patients, in order to positively influence postoperative outcome. PMID- 17410725 TI - [Human pancreas procurement for pancreas and islet transplantation]. AB - Insulin-dependent diabetes mellitus is a disease characterized by a deficiency of insulin secretion by beta islet cells. Integral pancreatic transplantation and islet cell transplantation represent two possibilities of replacing the beta islet cells. The aim of this paper is to analyze the main aspects of selection criteria of the pancreas donor and surgical methods of preservation for integral pancreas or islet cell transplantation. There are described the pancreas procurements realized by the Center of General Surgery and Liver Transplantation team from Fundeni Clinical Institute between December 2005-December 2006 and correlation between the donor's and pancreas features and the quality of islet cell isolation. The selection of pancreas donor and the accomplishment of pancreas procurement represent important factors in the post-transplantation course of the graft. The data from pancreas/islet cell transplantation centers suggest that the factors that influence positively the course of graft are: the donor's age, body mass index, cold ischemia time. PMID- 17410726 TI - [Fractal analysis in the diagnosis of breast tumors]. AB - Last years studies made by researchers from over the world show that fractal geometry is a viable alternative for image analysis. Fractal features of natural forms give to fractal analysis new valences in various fields, medical imaging being a very important one. This paper intend to prove that fractal dimension, as a way to characterize the complexity of a form, can be used for diagnosis of mammographic lesions classified BI-RADS 4, further investigations being not necessary. The experiments made on 30 cases classified BI-RADS 4 confirmed that 89% of benign lesions have an average fractal dimension under the threshold 1.4, meanwhile malign lesions are characterized, in a similar percentage, by an average fractal dimension over that threshold. PMID- 17410727 TI - [Laparoscopic surgery of hepatic hydatid cyst at the 3rd Surgical Clinic, Cluj Napoca]. AB - In the III-rd Surgical Clinic Cluj-Napoca, during 1996-2005, there was operated laparoscopic 33 patients with hepatic hydatid cyst. The sex ratio was M10/F23, with a mean age of 34 years and with ages between 10 and 66 years. Hepatic echography was used in preoperative diagnosis. The hepatic hydatid cysts were situated in 9 cases in the left liver (II-III-IV Couinaud segments) and in 24 cases in the right liver (15 cases in the V-VI segments and 9 cases in the VII VIII Couinaud segments). The laparoscopic treatment was performed by Lagrot pericystectomy at 31 patients and by ideal cystectomy at 2 patients. Around the cyst there were put switches impregnated with formalin 2% or hypertonic saline 20% to prevent peritoneal insemination. The cyst sterilization was done in most cases with hypertonic saline solution. The mean postoperative hospitalization was 5,6 days, with the range 1-21 days. Laparoscopic surgery with the well known advantages offers a good alternative to classic surgery in the treatment of hepatic hydatid cyst. PMID- 17410728 TI - [Surgical management of colonic diverticular disease]. AB - Diverticulosis is a common condition and affects one third of patients older than age 45. The treatment of the diverticular disease and of its complications, especially the acute inflammatory ones, is both medical and surgical. It is essential to correlate the two therapeutic means and to set an adequate time for the operation. The present study evaluates in a retrospective manner a number of 231 cases, 49% of which represent complicated forms. Fifty-eight patients underwent a surgical operation, 25 of which needed an emergency intervention. The results of the study show the increase of 1-stage procedures for the complicated forms and recommend laparoscopy as an alternative to open surgery. Recent studies have challenged the prophylactic nature of surgical operations after diverticulitis indicating a need for further evaluation. PMID- 17410729 TI - [Septic complications in acute appendicitis. Problems of diagnosis and treatment]. AB - AIM: diagnostic improvement in complicated acute appendicitis (AA) by implementing the new sepsis concepts and modern imaging procedures; optimization of treatment with decreasing postoperation morbidity and mortality and improving the cost-efficiency indicator. 1495 cases of AA admitted between 2000 and 2004 have been assessed retrospectively and among them 306 (20.46%) had complications. On admission 80.43% patients were diagnosed with AA, 17.50% with acute abdominal syndrome and 2.07% with chronic appendicitis. On discharge there were 1158 (77.45%) cases of inflammatory AA and 306 (20.46%) cases of complicated AA (perforation, gangrene, peritonitis, plastron abscess). TREATMENT: 98.26% of patients under-went operation and 1.73% did not. SURGICAL PROCEDURES: standard appendicectomy in 1407 (95.77%) cases; laparoscopic appendicectomy in 30 (2.04%) cases; extraperitoneal approach in 15 (1.02%) cases; associated with surgical interventions on other organs in 75 (5.10%) cases. POST-OPERATORY MORBIDITY:128 (41.08%) cases out of 306; septic parietal complications in 102 (33.33%) cases; intraperitoneal complications in 26 (8.49%) cases. POST-OPERATORY MORTALITY: 6 deaths in patients aged over 68. AVERAGE LENGTH OF STAY IN HOSPITAL: 4 days for uncomplicated AA and 14.6 days for complicated ones. Perioperative septic complications are the result of evolution of late diagnosed disease, unjustified postponing of operation, surgical technique, patient health condition. PMID- 17410730 TI - [Progress of surgical treatment in the last 10 years. Comparative study of surgical interventions performed between 1993-1995 versus between 2003-2005]. AB - In the last decade of the past century, as laparoscopy was introduced in our clinic in 1993, minimal access therapy (MAT--endoscopy, angiography, interventional imagery) had a positive and constant evolution. Our paper retrospectively evaluates the interventions performed between 2003-2005 (group A) compared to those performed between 1993-1995 (group B). We observed a 17.08% (7056 vs 6026 interventions/year) raise in the total number of interventions in group A, with a significant 66% decrease (195 vs. 588 interventions/year) of interventions for gastro-duodenal ulcer and a 18% decrease (1211 vs 1490 interventions/year) of appendectomies, but a 63% increase (1560 vs. 955 interventions/year) of cholecystectomies, 53% increase (1186 vs. 773 interventions/year) of interventions for parietal defects and a 62% (626 vs. 325 interventions/year) increase of oncological interventions. The most frequent interventions were, in the order of frequencies: cholecystectomies (79.8% laparoscopically), appendectomies, interventions for hernia and eventrations, oncological operations and trauma surgery. The incidence of laparoscopic interventions was greater in group A, counting for 19% of the total number of interventions. In group A were performed 2334 endoscopies, 149 diagnostic and therapeutic angiographies. Postoperative mortality dropped with 29.64% and hospital stay dropped to 4.7 days. We believe that the incidence of MAT should rise, by performing more laparoscopic interventions and this change should lead to a revision of the surgical residents training program. PMID- 17410731 TI - [Experimental study of the use of piritramide as intrathecal analgesic]. AB - Opioids proved their advantages as general and intrathecal (i.t.) analgesics. Piritramide (P), a largely used analgesic opioid today, has not been studied in i.t. administration. Our experimental research aimed in determining the efficiency, security and optimal dose of i.t. P. In 9 adult mongrel dogs equally randomized in 3 groups we injected i.t. P 1.3 mg x kg-l (group 1), P 0.8 mg x kg l (group 2) and sodium chloride 0,9% (group 3) and we registered the motility, the pain reaction to electrical and mechanical nociceptive stimuli, the respiratory rate and amplitude, electrocardiogram, heart rate, mean arterial blood pressure electroencephalogram and, for 2 subjects from group 1, electromyogram. The P-induced analgesia was strong, dose-dependent, and segmental, with a time of onset of 5-8 min, duration of 1h 45 min-2h 30 min, and prolonged residual analgesic level for 5-6 h. The dogs from the 1st group presented moderate side effects: bradypnea, tachycardia and arterial hypotension at 5 min, reduction in the posterior limbs motility, sleep. We could conclude that i.t. piritramide 0.8 mg x kg-l provides a solid, segmental, long-lasting analgesia, without marked adverse effects. PMID- 17410732 TI - [Peritoneal biofilms: microscopic features]. AB - Antibiotherapy remains one of the basic clinical tools, which can influence the evolution of severe peritonitis. Peritoneal biofilm formation may minimize the antibiotic effects due to dramatic growth of Minimum Inhibitory Concentrations (MIC) and Minimum Bactericidal Concentrations (MBC) for matrix-enclosed bacteria. In this paper we demonstrate the presence and evolution of bacterial biofilms on the peritoneal surface during the course of severe secondary peritonitis using an experimental model and clinical material. Cecal Ligation Puncture was performed in 20 mice Swiss Webster. Peritoneal samples were studied at optic and electronic microscope at 10, 24, 48 and 72 hours postoperative. Clinical samples were taken from 10 patients with diffuse peritonitis. At 24 hours after the onset of the peritonitis bacterial colonies were detected on the peritoneal surface. The formation of mature multilayer polymicrobial biofilms with deep penetration in abdominal wall by 48-72 hours was documented. The bacterial biofilms appear in first 24 hours in the course of experimental generalized peritonitis. Our experimental and clinical data demonstrate formation of the mature polymicrobial biofilm in 48-72 hours after the onset of peritonitis. The possibility of resistant biofilm formation in secondary diffuse peritonitis should be taken into consideration in elaboration of treatment schemes. PMID- 17410733 TI - [Technique of internal surgical stabilization for flail chest]. PMID- 17410734 TI - Laparoscopic hand-assisted adrenalectomy for a 20 cm benign tumor. AB - Since its introduction in 1992, laparoscopic adrenalectomy (LA) has become the technique of choice in the surgical treatment of both secreting or non-secreting benign adrenal pathology. Although traditionally, laparoscopic approach was recommended only for tumor sizes less than 6-8 cm--as larger tumors were known to have an increased risk of malignancy--the currently growing experience and improvement of surgical techniques has allowed for an extension of the therapeutic indication, as shown by the recent case report of LA use for a benign 22 cm tumor (1). We report the case of a young patient operated in our Department for a benign 20 cm adrenal tumor for which laparoscopic "hand-assisted" adrenalectomy yielded a good postoperative outcome and minimal complications. PMID- 17410735 TI - [Viral small bowel intussusception: a rare cause of intestinal obstruction]. AB - Representing a rare cause of bowel obstruction, the ileal intussusception is commonly met in the pediatric surgery. Even if in children's cases the symptoms can mimick a multitude of abdominal syndromes, usually in adult cases the symptoms fit the pattern of the intestinal obstruction. This paper presents 2 clinical cases of small bowel intussusception in adult, the particularity of cases being that the pathogenesis couldn't be established first hand; the pathology exam revealed only minor inflammatory responses,including modest reactive lymph nodes in the vicinity of lesions, without further alterations. The etiology of bowel intussusception was finally attributed to viral infection with gastroenteritis, based on clinical and pathological criteria. PMID- 17410736 TI - [Pancreas divisum--a rare cause of chronic pancreatitis]. AB - Pancreas divisum (P.D.) is a congenital anatomic variant, characterized by the nonunion of dorsal and ventral pancreatic ducts. A 20 years old man followed for 8 years with reccurent abdominal pain and relapsing acute pancreatitis develope chronic calcific pancreatitis. He was diagnosed with P.D. on endoscopic retrograde pancreatography and operative pancreatography. The patient was treated with longitudinal pancreatico-jejunostomy (PUESTOW-GILLESBY procedure). His pain resolved following surgical drainage of the pancreatic duct. Evaluation of the clinical course of this patient and critical review of other such cases in the literature support the role of compromised ductal drainage of the pancreas in the pathogenesis of chronic pancreatitis in P.D. PMID- 17410737 TI - [Hynes-Andersen pyeloplasty by mini-laparoscopy transperitoneal approach]. AB - Laparoscopy had significantly gained in the field of urologic surgery, almost all the surgical procedures in the retroperitoneal space being made, from the simple ones (renal cystectomy, varicocelectomy) to the ones requiring higher expertise (nephrectomy, adrenalectomy) by retro-peritoneoscopy and also transperitoneal. At the Surgery Department of "Sf. Ioan" Hospital, Bucharest, we approached initially the retroperitoneal pathology by lomboscopy, but afterwards we choose the transperitoneal approach because of the generous workspace and the clearer anatomical landmarks, regardless of the procedures (renal cystectomy, varicocelectomy, adrenalectomy, nephrectomy, pyelo-lithotomy, ureterolithotomy). In association with the Urology Department of "Sf. Ioan" Hospital we practiced two Hynes Andersen pyeloplasty for pyelo-ureteral junction stenosis by transperitoneal approach, using mini-laparoscopy tools. In one of the cases we also perform a basinetal lithotomy. PMID- 17410738 TI - Primary torsion of the greater omentum. An obscure and unusual cause of acute abdomen. AB - A 36-year-old man was admitted with a 3-day history of severe abdominal pain in the right upper abdomen and was initially diagnosed with acute cholecystitis or acute retrocecal appendicitis. The patient was transferred to the department of surgery for close surgical observation. CT of the entire abdomen was performed just before the operation, which demonstrated inflammation in the omental fat. Surgery revealed primary omental torsion and subsequent resection of the infarcted segment offered a rapid recovery. We report a case of primary segmental omental torsion and discuss the diagnostic and therapeutic implications of this unusual entity. PMID- 17410739 TI - Concurrent low grade B-cell non-Hodgkin's lymphoma of MALT type arising in the large intestine, small intestine and stomach. AB - Low-grade non-Hodgkin's lymphomas (NHL) of mucosa associated lymphoid tissue (MALT) are indolent neoplasms that, although tending to remain localized for many years, may spread to other mucosal sites. Despite increasing identification of concurrent gastric and intestinal lymphoma of MALT type, the clonal relationship between the tumors and their sequential development are poorly understood. It is also unknown whether the development of these concurrent tumors is closely associated with direct antigen stimulation, which is thought to play an important role in the clonal expansion of low grade MALT lymphomas. The most important function of B-cells is production of specific antibodies. This is largely achieved during B-cell development by recombination of the Ig heavy chain variable (V), diversity (D), and joining (J) segments and hypermutation of the rearranged gene. The rearranged Ig genes of a mature B-cell record much of its evolution history. We report a case of synchronous development of intestinal and gastric low grade MALT lymphomas in a 70 years old female and discuss their possible clonal relationship and sequential appearance. PMID- 17410740 TI - [Fungal native aortic valve endocarditis. Combined medico-surgical treatment]. AB - The fungal etiology of the native aortic endocarditis is a rare situation. Most often the etiologic factors are medical manoeuvres like cardiovascular surgery (cardiac prosthesis), cardiac defibrillators or stimulators, the prolonged use of the intravenous catheters, antibiotics, immunosuppressors, etc. We are presenting the case of a patient with cardiac stimulator, with rare fungal native aortic endocarditis, complicated with cerebral embolization, in which we have chosen a combined therapy: aortic replacement with biologic prosthesis and pre and post operative anti fungal therapy. We are considering that in this case the combined medico-surgical therapy was the right choice. PMID- 17410741 TI - Choledochoduodenal anastomosis: a technical tip to ensure the impermeability of the anastomosis. AB - In 1891 O. Sprengel described a successful side-to-side choledochoduodenostomy and since then this has been a method frequently used in dealing with the problem of constriction of the lower third-portion of the bile duct, due to malignant or benign causes. Taking in consideration that the first choledochoduodenostomy was performed by H. Riedel in 1888 and the patient died 9 hours after surgery due to leakage of the anastomosis (disclosed in autopsy), it is evident to conclude that hermetic anastomosis is the main factor for fast and uncomplicated recovery of the patient. In order to ensure intraoperatively a hermetic "bile-proof" anastomosis, we have applied a technical tip, which has been proved to be efficient. PMID- 17410743 TI - [National Congress of Surgery, Serbia. Novi Sad, November 23-24, 2006]. PMID- 17410744 TI - [The Third International Interactive Conference of current state in hepatic transplant surgery. Hospital Niguarda, Milano, December 14-15, 2006]. PMID- 17410745 TI - [The 14th Symposium and Post-University International Course of IASGO, (International Association of Surgeons, Gastroenterologists and Oncologists. Athens, December 7-9, 2006]. PMID- 17410746 TI - [1st Romanian-Serbian Surgical Conference, Drobeta-Turnu Severin, November 17-19, 2006]. PMID- 17410747 TI - Navigator role shows promise in decreasing cancer death rates for all populations. PMID- 17410748 TI - Taking the wheel. Oncology nurses help patients navigate the cancer journey. PMID- 17410750 TI - What nursing interventions improve fatigue in patients with cancer? PMID- 17410749 TI - How do you make a difference in patients' lives through your role as a nurse navigator? PMID- 17410751 TI - Navigators: the champions of finding solutions to healthcare barriers. PMID- 17410752 TI - Re-defining the term health disparities. PMID- 17410753 TI - The Adolescent Lifestyle Profile: development and psychometric characteristics. AB - Adolescents establish patterns of behavior and make lifestyle choices that affect their future health during their transition from childhood to adulthood. They struggle with behaviors, such as physical activity and nutrition, which will affect their risk of developing chronic diseases in adulthood. A comprehensive, easy to administer instrument is needed that is both research worthy and clinically useful in order to assess adolescent lifestyle behaviors and to plan interventions appropriately. The purpose of this paper is to report the development and testing of the Adolescent Lifestyle Profile (ALP), a Likert-type instrument to measure seven domains of a health-promoting lifestyle in adolescents. The ALP was modeled after the Health Promoting Lifestyle Profile II (HPLP II) and tested in a sample of early adolescents. Internal consistency reliability, including Cronbach's alpha, item to total correlations and subscale to total scale correlations and construct validity, including concurrent validity testing and factor analysis, indicated that the ALP is a reliable and valid scale that can be used to assess healthy lifestyle domains in adolescents. PMID- 17410754 TI - Legume intake and reduced colorectal adenoma risk in African-Americans. AB - Colorectal adenomas are known precursors for colorectal cancer. Several studies have shown that dietary factors can influence adenoma formation and growth. This study was conducted using African-American men and women who were undergoing colonoscopies in order to examine the relationship between selected dietary factors and the risk for colon polyps. In a case-control design, 186 men and women with a mean of 58 years of age were studied. A multiple logistic regression model was used to adjust for potential confounding variables and to determine which factors influence colorectal adenoma risk. Study results revealed that consumption of legumes such as dried beans, split peas, or lentils was negatively associated with risk (OR = 0.19; 95% CI: 0.04-0.91). Legumes are a good source of dietary fiber and of phytochemical compounds that may play a role in reducing adenoma formation or growth, thereby decreasing the risk of colorectal cancer. Nurses working with African-Americans should encourage consumption of these foods to decrease this risk. PMID- 17410755 TI - Health belief model perceptions, knowledge of heart disease, and its risk factors in educated African-American women: an exploration of the relationships of socioeconomic status and age. AB - Heart disease is the leading cause of death for African-American women in the United States. Although African-American women experience higher rates of heart disease with earlier onset and more severe consequences than White women do, they are not aware of their risk for the disease. The Health Belief Model (HBM) has been commonly used to guide preventive interventions in cardiovascular health. However, the HBM has not been evaluated for African-American women regarding its effectiveness. This study explored the perceptions of susceptibility and seriousness of heart disease, and the relationships between socioeconomic status (SES), age, and knowledge of heart disease and its risk factors among 194 educated African-American women from the southern United States. Participants did not perceive themselves to be at high risk for developing heart disease while perceiving heart disease as serious. African-American women who were older perceived heart disease to be more serious than their younger counterparts did. Older women and those with higher SES knew more about heart disease and risk factors. Neither SES nor age moderated the relationship between knowledge and perceived susceptibility or seriousness. PMID- 17410756 TI - Nursing students' adjustment to a new phenomenon. AB - The purpose of this article is to examine how undergraduate nursing students are adjusting to the Web learning environment. By learning how these students are adjusting to the Web environment, the Web can be tailored to meet their needs. The article examined how the nursing students felt about their access to the Web environment, their interactions with faculty and peers, their overall experience in the Web environment, and the Web course itself. The findings from this study were that the undergraduate nursing students had a positive experience in the Web environment. Suggestions made by the students included providing technical support during evening hours and having someone on call to address technical issues during this time. The students also suggested more or different office hours on the Web for the instructors and the utilization of chat rooms to provide support and answer questions. PMID- 17410757 TI - African-American adolescent females' predictors of having sex. AB - African-American adolescent females are at high risk for HIV infection, acquired primarily as a result of heterosexual intercourse. Multiple regression analyses was used to describe the correlates of the outcome variable, ever having had sex, for low-income African-American adolescent females in a cross-sectional study of 322 (N = 322) mother/daughter pairs. The results revealed that while the daughters' age was positively associated with the outcome variable, the daughters' perceptions of strictness of curfew and maternal monitoring, and the daughters'self-efficacy to refuse sex and their intention to refuse sex were negatively associated with the outcome variable. Mothers tended to report significantly more maternal monitoring and stricter curfews than what their daughters perceived them to be. The daughters'intention to refuse sex mediated the effects of age and self-efficacy to refuse sex on the outcome variable. These results suggest that health-care providers should promote clearer communication between mothers and daughters in order to reduce divergent perceptions and to help mothers facilitate their daughters' self-efficacy and their intention to refuse sex. PMID- 17410758 TI - Women and HIV/AIDS in the kingdom of Swaziland: culture and risks. AB - In Swaziland, a polygamous society in Southern Africa, the prevalence of the human immune virus/acquired immune deficiency syndrome (HIV/AIDS) is continuing to proliferate at an alarming rate. In 1992 the prevalence rate was 3.9%. However in 12 years, by 2004, the prevalence rate had reached 42.6%. This article explores some of the traditional cultural practices and experiences that increase Swazi women's vulnerability to HIV/AIDS. The traditional cultural practices fall into four categories: (1) socialization and the roles of women, (2) the minority status of women, (3) the practice of a dowry, and (4) the wife as an inheritance. The women's experiences include the Swazi men's beliefs in the virginity cure myth, the women's extreme poverty, and the Swazi men who are migrant workers in neighboring states. This article concludes with recommendations for public policy and for future research within the context of Swazi culture. PMID- 17410759 TI - An overview and update on the metabolic syndrome: implications for identifying cardiometabolic risk among African-American women. AB - Identifying patients with cardiovascular risks was once the simple process of recognizing the traditional risk factors. However, since the National Cholesterol Educational Program Adult Treatment Panel III guidelines were released in 2001, and again revised in 2004, identification of cardiovascular risk has become more complicated. This article provides both an overview and an update on the metabolic syndrome as a cluster of risk factors for both type 2 diabetes and cardiovascular disease (CVD). Specific issues are discussed as they relate to African-American women and their prevalence of the metabolic syndrome and cardiometabolic risk. Nurses are optimally prepared to identify the early signs of cardiometabolic risk by testing for and assessing the metabolic syndrome. Likewise, nursing is in a position to provide teaching and counsel regarding the needed lifestyle alterations for patients with metabolic syndrome in order to reduce the risk of type 2 diabetes, cardiovascular disease, and/or stroke. PMID- 17410760 TI - Dry deposition of mercury. PMID- 17410761 TI - Mixed news on Chinese farmed fish. PMID- 17410763 TI - Will REACH serve researchers' needs? PMID- 17410762 TI - Record-high PBDEs in Chinese birds of prey. PMID- 17410764 TI - U.S. EPA trims the public's right to know. PMID- 17410765 TI - World's first low-carbon fuel provision. PMID- 17410767 TI - European body for environmental chemists. PMID- 17410766 TI - U.S. EPA to revisit asbestos toxicity. PMID- 17410768 TI - Contaminated salmon and the public's trust. PMID- 17410769 TI - Simultaneous assessment of deposition effects of base cations, sulfur, and nitrogen using an extended critical load function for acidification. AB - Base cations (BC) play an important role to prevent soil acidification. In certain acid sensitive areas, such as China, BC deposition is high and a considerable fraction is of anthropogenic origin. BC deposition might decrease in the future with the implementation of air pollution control measures. The effect of changes in BC deposition, however, has seldom been considered in critical load calculations based on the steady-state mass balance (SSMB) method. In order to better quantify the importance of the BC deposition for acid deposition mitigation policy, an extension of the SSMB method for critical load calculation for soil acidification is presented. The BC deposition is taken into account as a variable along with sulfur (S) and nitrogen (N) deposition, creating an S-N-BC critical load function. As a case study, critical loads of S and N for the Tie Shan Ping catchment in Chongqing in southwest China under variable BC deposition were calculated. Results indicate that abatement of BC deposition has significant impact on the critical loads of S and N. A 75% reduction in BC of assumed anthropogenic origin decreases the critical loads of acids by 58%. The current deposition does not exceed the critical loads, but if BC deposition from anthropogenic sources was controlled, then the exceedance would be considerable. Uncertainty analysis show that the size of the BC deposition of natural origin is the single parameter contributing the most to the steady-state S and N critical load. The extended critical load function can be used by policy makers to set more reasonable acidity control strategies in the future. The method also highlights for policymakers the "competition" between emission control of particulate matter driven by human health targets and potential increase of net acid load from such measures. PMID- 17410770 TI - Persistent halogenated hydrocarbons in consumer fish of China: regional and global implications for human exposure. AB - Concentrations of persistent halogenated hydrocarbons (PHHs) were determined in 13 consumer fish species (a total of 390 individuals) collected from a major fish farming region of China. The potential health risks of PHHs associated with consumption of fish from China was also systematically assessed regionally and globally. In all fish samples, DDTs, HCHs, PCBs, and PBDEs were the predominant PHH residues, with the median levels (ranges) being 6.0 (0.14-698.9), 0.50 (0.13 24.06), 0.10 (< 0.02-7.65), and 0.15 (< 0.0012-3.85) ng/g wet weight, respectively. The upper-bound (90th percentile) values of estimated daily intakes of DDT, HCHs, PCBs, and PBDEs via fish consumption were 45.5, 1.35, 0.46, and 0.30 ng/kg bw/d (urban), and 15.9, 0.47, 0.16, and 0.10 ng/kg bw/d (rural). Globally, the upper-bound outflows via fishery exportation of DDT, HCHs, PCBs, and PBDEs were 185, 5.51, 1.86, and 1.22 kg, respectively, in 2005. Japan was the largest recipient of PHHs, followed by Korea and the United States. Fish consumption assessments indicated that consumption of freshwater farmed and wild marine fish generally does not subject consumers to significant health risk as far as PHHs are concerned, while limited consumption of seawater farmed fish is advised. PMID- 17410771 TI - Polybrominated diphenyl ethers in birds of prey from Northern China. AB - Birds of prey from Northern China (Beijing area) were examined for polybrominated diphenyl ethers (PBDEs). A total of 47 specimens from eight different species were analyzed. Muscle and liver were analyzed separately for each bird. Kidneys were pooled by species. Common kestrels exhibited the highest PBDE levels (mean muscle and liver concentrations of 12300 and 12200 ng/g lipid weight, respectively), with maxima in an individual bird of 31700 in muscle and 40900 ng/g lw in liver. Congener profiles differed between some species, but were generally dominated by the more brominated congeners (e.g., BDE-153, -209, -183, 207). BDE-209 was especially elevated compared to other published reports. Interspecies differences in congener concentrations and profiles may be due to diet, behavior, or biotransformation capacities. BDE-209 was detected in 79.4% of the samples. Common kestrels contained the highest BDE-209 levels (mean/maxima of 2150/6220 in muscle and 2870/12200 ng/g lw in liver). BDE-209 was the dominant congener in tissues from some buzzards, scops owls, and long-eared owls. It was the second most abundant congener in common kestrels. The remarkable levels and dominance of BDE-209 may relate to significant production, usage, or disposal of deca-containing products in China. These observations reinforce the growing view that organisms using terrestrial food chains may have greater exposure to BDE 209. PMID- 17410772 TI - Hibernation-associated changes in persistent organic pollutant (POP) levels and patterns in British Columbia grizzly bears (ursus arctos horribilis). AB - We hypothesized that depleted fat reserves in grizzly bears (Ursus arctos horribilis) following annual hibernation would reveal increases in persistent organic pollutant (POP) concentrations compared to those present in the fall. We obtained fat and hair from British Columbia grizzly bears in early spring 2004 to compare with those collected in fall 2003, with the two tissue types providing contaminant and dietary information, respectively. By correcting for the individual feeding habits of grizzlies using a stable isotope-based approach, we found that polychlorinated biphenyls (sigmaPCBs) increased by 2.21x, polybrominated diphenylethers (sigmaPBDEs) increased by 1.58x, and chlordanes (sigmaCHL) by 1.49x in fat following hibernation. Interestingly, individual POPs elicited a wide range of hibernation-associated concentration effects (e.g., CB 153, 2.25x vs CB-169, 0.00x), resulting in POP pattern convergence in a PCA model of two distinct fall feeding groups (salmon-eating vs non-salmon-eating) into a single spring (post-hibernation) group. Our results suggest that diet dictates contaminant patterns during a feeding phase, while metabolism drives patterns during a fasting phase. This work suggests a duality of POP-associated health risks to hibernating grizzly bears: (1) increased concentrations of some POPs during hibernation; and (2) a potentially prolonged accumulation of water soluble, highly reactive POP metabolites, since grizzly bears do not excrete during hibernation. PMID- 17410773 TI - Detection of hydroxylated polychlorinated biphenyls (OH-PCBs) in the abiotic environment: surface water and precipitation from Ontario, Canada. AB - Hydroxylated PCBs (OH-PCBs) are well-known metabolites of PCBs in organisms, but there has been no direct study of their presence in the abiotic environment. In this study, OH-PCBs were determined in samples of rain, snow, and surface waters from sites in Ontario, Canada. OH-PCBs were quantified by gas chromatography with high-resolution mass spectrometry (GC-HRMS)in order to provide complete characterization of all OH-PCB homolog groups. OH-PCBs and PCBs were detected in all the samples analyzed, although half of the sigmaOH-PCBs could not be identified even with 71 individual congener standards. Total concentrations of OH PCBs (sigmaOH-PCBs) in water ranged from 0.87 to 130 pg/L and from 230 to 990 pg/g in particulate organic matter. Total fluxes of those compounds in snow and rain were from < 1 to 100 pg/m2 and from < 1 to 44 pg/ m2/day, respectively. Higher sigmaOH-PCB fluxes in rain were found in southern Ontario than in a remote north-central Ontario site possibly reflecting greater sources of precursor PCBs near urban areas. Relatively higher sigmaOH-PCB concentrations were found in surface waters from sites near sewage treatment plant (STP) outfalls in the cities of Toronto (130 pg/L) and Hamilton (35 pg/L) than in offshore samples from Lake Ontario (1.6 pg/L). The results indicate that STPs are one of the sources of OH-PCBs for lake waters in this region. Similar homolog and congener profiles in rain and offshore surface water samples suggest that atmospheric deposition is the predominant source at offshore sites. This is the first report to detect the OH-PCBs in the abiotic environment. PMID- 17410774 TI - Predicting effects of exploitation rate on weight-at-age, population dynamics, and bioaccumulation of PCDD/Fs and PCBs in herring (Clupea harengus L.) in the Northern Baltic Sea. AB - The Baltic Sea ecosystem and fish stocks contain high concentrations of environmental chemicals such as polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs), and polychlorinated biphenyls (PCBs). This study forecasts how changes in fishing or natural mortality would probably influence concentrations of PCDD/F and PCB in the Bothnian Sea (Northern Baltic) herring (Clupea harengus L.). An age-structured simulation model was developed to forecast herring stock dynamics, catches, and weight-at-age under different assumptions about exploitation and natural mortality. The simulated herring weight-at-age estimates were employed in a bioenergetics model capable of simultaneous estimation of bioaccumulation of 17 PCDD/F and 37 PCB congeners. Although the natural variability in recruitment greatly influences the stock dynamics, considerable changes in weight-at-age would ensue changes in exploitation rate or in natural mortality rate. If exploitation rates increase, growth rates would be higher and herring in the weight categories of commercial fisheries would be younger and contain less PCDD/F and PCB. Hence, the average toxicant concentrations in catches would also decline. However, it is likely that only fairly small changes would occur in toxicant concentrations-at-age. On the other hand, a drastic decrease in herring fishing would substantially increase PCDD/F and PCB concentrations in herring. The study indicated that, in spite of the clear influences of fishing on the toxicant concentrations, fishing alone cannot resolve the problems associated with a high concentration of toxicants in herring; further decreases in loading are still required. PMID- 17410775 TI - Detection of Bacteroidales fecal indicators and the zoonotic pathogens E. coli 0157:H7, salmonella, and campylobacter in river water. AB - Bacteroidales host-specific PCR offers a rapid method of diagnosing fecal pollution in water and identifying sources of input. To assess human health risks from exposure to fecal pathogens, however, Bacteroidales markers should be detectable when pathogens are present. To determine if Bacteroidales general, human-, ruminant-, and swine-specific markers correlate with certain fecal pathogens, we conducted a retrospective study on water samples for which the presence of E. coli O157:H7, Salmonella spp., and Campylobacter spp. had been determined. We found a positive relationship between detection of the Bacteroidales general fecal marker and presence of the pathogens. Detection of ruminant-specific markers predicted E. coli O157: H7 occurrence. There was a significant increase in the likelihood of detecting Salmonella when a ruminant marker was present, and Campylobacter spp. when human markers were present. For pathogens such as E. coli O157: H7 that are strongly associated with particular hosts, Bacteroidales host-specific markers can estimate the likelihood of pathogen occurrence, enabling more accurate health risk assessments. PMID- 17410776 TI - Selenium volatiles as proxy to the metabolic pathways of selenium in genetically modified Brassica juncea. AB - In this study we demonstrate that the headspace selenium volatiles could be used as proxy to the metabolic pathways in the Se-accumulator plant Brassica juncea. The selenium metabolic pathways in wild type plants are compared to those of several genetically modified cultures. Complementary use of atomic and molecular mass spectrometric techniques also allowed for identification of yet unreported minor headspace Se-containing volatiles such as CH3SeSeSeCH3, CH3SeSSeCH3, and CH3SeCH2CH3. By combining the information resulting from this research with the previously known information about selenium metabolism in B. juncea, it is possible that a more efficacious phytoremediation tool can be constructed. PMID- 17410777 TI - Stable hydrogen and oxygen isotope composition of waters from mine tailings in different climatic environments. AB - The stable isotope composition of waters (delta2H, delta18O) can be used as a natural tracer of hydrologic processes in systems affected by acid mine drainage. We investigated the delta2H and delta18O values of pore waters from four oxidizing sulfidic mine tailings impoundments in different climatic regions of Chile (Piuquenes at La Andina with Alpine climate, Cauquenes and Caren at El Teniente with Mediterranean climate, and Talabre at the Chuquicamata deposit with hyperarid climate). No clear relationship was found between altitude and isotopic composition. The observed displacement of the tailings pore waters from the local meteoric water line toward higher delta18O values (by approximately +2 per thousand delta18O relative to delta2H) is partly due to water-rock interaction processes, including hydration and O-isotope exchange with sulfates and Fe(III) oxyhydroxides produced by pyrite oxidation. In most tailings, from the saturated zone toward the surface, isotopically different zones can be distinguished. Zone I is characterized by an upward depletion of 2H and 18O in the pore waters from the saturated zone and the lowermost vadose zone, due to ascending diffused isotopically light water triggered by the constant loss of water vapor by evaporation at the surface. In zone II, the capillary flow of a mix of vapor and liquid water causes an evaporative isotopic enrichment in 2H and 18O. At the top of the tailings in dry climate a zone III between the capillary zone and the surface contains isotopically light diffused and atmospheric water vapor. In temperate climates, the upper part of the profile is affected by recent rainfall and zone III may not differ isotopically from zone II. PMID- 17410778 TI - Reduction of nickel and uranium toxicity and enhanced trichloroethylene degradation to Burkholderia vietnamiensis PR1301 with hydroxyapatite amendment. AB - The use of hydroxyapatite (HA) to sequester metals at mixed waste sites may reduce metal toxicity and facilitate microbial degradation of cocontaminant organics. The constitutive trichloroethylene (TCE) degrader, Burkholderia vietnamiensis PR1301, grew at 34.1 and 1.7 mM Ni at pH 5 and 7, respectively, with 0.01 g mL(-1) HA compared to 17 and 0.85 mM Ni without HA. PR1 grew at 4.2 mM U at pH 5 and 7 with 0.01 g mL(-1) HA compared to 1.1 mM U without HA. A similar decrease in the toxicity of Ni and U in combination was observed with HA. The ability of PR1 to degrade TCE at 0.85, 1.7, and 3.4 mM Ni and at 0.42 and 1.1 mM U was examined. The presence of TCE resulted in a decreased tolerance of PR1 to Ni and U; however, HA facilitated TCE degradation in the presence of Ni and U, effectively doubling the metal concentrations at which TCE degradation proceeded. These studies suggest that metal sequestration via HA amendments may offer a feasible approach to reducing metal toxicity to microorganisms at mixed waste sites, thereby enhancing the degradation of cocontaminant organics. PMID- 17410779 TI - Biological and chemical redox transformations of mercury in fresh and salt waters of the high arctic during spring and summer. AB - It is well-established that atmospheric deposition transports Hg to Arctic regions, but the postdepositional dynamics of Hg that can alter its impact on Arctic food chains are less understood. Through a series of in situ experiments, we investigated the redox transformations of Hg in coastal and inland aquatic systems. During spring and summer, Hg reduction in streams and pond waters decreased across a 4-fold increase in salinity. This alteration of Hg reduction due to chloride was counterbalanced by the presence of particles, which favored the conversion of oxidized Hg to its elemental form. In saline waters, biogenic organic materials, produced by algae, were able to promote oxidation of Hg(O) even under dark conditions. Overall these results point to the vulnerability of marine/ coastal Arctic systems to Hg, compared to inland systems, with oxidation processes enhancing Hg residence times and thus increasing its potential to enter the food chain. PMID- 17410780 TI - Mercury stable isotope fractionation during reduction of Hg(II) to Hg(0) by mercury resistant microorganisms. AB - Mercury (Hg) undergoes systematic stable isotopic fractionation; therefore, isotopic signatures of Hg may provide a new tool to track sources, sinks, and dominant chemical transformation pathways of Hg in the environment. We investigated the isotopic fractionation of Hg by Hg(II) resistant (HgR) bacteria expressing the mercuric reductase (MerA) enzyme. The isotopic composition of both the reactant Hg(II) added to the growth medium and volatilized product (Hg(0)) was measured using cold vapor generation and multiple collector inductively coupled plasma mass spectrometry. We found that exponentially dividing pure cultures of a gram negative strain Escherichia coli JM109/pPB117 grown with abundant electron donor and high Hg(II) concentrations at 37, 30, and 22 degrees C, and a natural microbial consortium incubated in natural site water at 30 degrees C after enrichment of HgR microbes, preferentially reduced the lighter isotopes of Hg. In all cases, Hg underwent Rayleigh fractionation with the best estimates of alpha202/198 values ranging from 1.0013 to 1.0020. In the cultures grown at 37 degrees C, below a certain threshold Hg(II) concentration, the extent of fractionation decreased progressively. This study demonstrates mass-dependent kinetic fractionation of Hg and could lead to development of a new stable isotopic approach to the study of Hg biogeochemical cycling in the environment. PMID- 17410781 TI - Fractionation of stable iron isotopes in higher plants. AB - Although the fractionation of stable iron isotopes by biological processes in the environment is currently a matter of intense debate, the isotope fractionation associated with the growth of higher plants has, to date, not been characterized. We show that iron isotope fractionation induced by higher plants is substantial and also generates systematic plant-specific patterns. We suggest a hypothesis in which these patterns mirror the two different strategies that plants have developed to incorporate iron from the soil: reduction of Fe(III) in soils by strategy I plants results in the uptake of iron, which is depleted in 56Fe by up to 1.6 per mil relative to 54Fe when compared to the available Fe in soils; complexation with siderophores by strategy II plants results in the uptake of iron that is 0.2 per mil heavier than that in soils. Furthermore, younger parts of strategy I plants get increasingly depleted in heavy isotopes as the plant grows, while strategy II plants incorporate nearly the same isotope composition throughout. This points to entirely different translocation mechanisms between strategy I and II plants. Such presumably redox-related differences in translocation have been under debate up to now. We conclude that plant metabolism represents an important cause of isotopic variation in the biogeochemical cycling of Fe. Therefore, heavy stable metal isotope systems now start to be viable indicators of geosphere-biosphere metal transfer processes. PMID- 17410782 TI - Co speciation in hardened cement paste: a macro- and micro-spectroscopic investigation. AB - Cement-based materials play an important role in multi-barrier concepts developed worldwide for the safe disposal of hazardous and radioactive wastes. Cement is used to condition and stabilize the waste materials and to construct the engineered barrier systems (container, backfill and liner materials) of repositories for radioactive waste. In this study, Co uptake by hardened cement paste (HCP) has been investigated with the aim of improving our understanding of the immobilization process of heavy metals in cement on the molecular level. X ray-absorption spectroscopy (XAS) on powder material (bulk-XAS) was used to determine the local environment of Co in cement systems. Bulk-XAS investigations were complemented with micro-beam investigations to probe the inherent microscale heterogeneity of cement by using micro-X-ray-fluorescence (micro-XRF) and micro XAS. Micro-XRF was used to gain information on the spatial heterogeneity of the Co distribution, whereas micro-XAS was employed to determine the speciation of Co on the microscale. The Co-doped HCP samples hydrated for time-scales from 1 hour up to 1 year were prepared under normal atmosphere, to simulate similar conditions as for waste packages. To investigate the role of oxygen, further samples were prepared in the absence of oxygen. The study showed that, for the samples prepared in air, Co(II) is oxidized to Co(III) after 1 hour of hydration time. Moreover, the relative amount of Co(III) increases with increasing hydration time. The study further revealed that Co(II) is predominately present as a Co-hydroxide-like phase and/or Co-phyllosilicates, whereas Co(III) tends to be incorporated into a CoOOH-like phase and/or Co-phyllomanganates. In contrast to samples prepared in air, XAS experiments with samples prepared in the absence of oxygen revealed solely the presence of Co(II). This finding indicates that oxygen plays an important role for Co oxidation in cement. Furthermore, the study suggests that Co(III) species or Co(III)-containing phases should be taken into account for an overall assessment of the Co release from Co-containing cement stabilized waste under oxidizing conditions. PMID- 17410783 TI - Identification and characteristics of a cyanobacterium isolated from a hot spring with dissolved inorganic carbon. AB - A photosynthetic species, isolated from an alkaline hot spring in eastern Taiwan, was applied to enhance the dissolved inorganic carbon (DIC) uptake capability under high alkaline and temperature conditions and was identified. The strain was found to be close to Thermosynechococcus elongatus BP-1 and Synechococcus elongatus based on phylogenetic analysis of 16S rRNA genes. The result was verified with morphological observations. This strain was named as Thermosynechococcus sp. CL-1 (TCL-1). A study of the effects of pH and DIC on the content variations of four elements (C, N, H, and O), lipids (LI), proteins (PR), carbohydrates (CA), and the bioenergy potential was carried out. The concentrations of PR and LI were the highest under the cultivation of pH 7 and CA was at 10.5. According to the analysis of three compositions, the production pathway of LI might be shifted to CA from pH 7 to 10.5 and then shifted to inorganic compounds from pH 10.5 to 11. Regarding the effect of DIC at pH 9, the results revealed that the uptake pathway shift (such as metals uptake) might happen while DIC is less than 18.9 mM. From 18.9 to 47.2 mM DIC, the production pathway of LI shifted to CA and the contents of CA increased quickly from 47.2 to 94.3 mM without a further decrease of LI. Regarding the pyrolysis experiments with a thermogravimetric analyzer coupled with Fourier transform infrared spectroscopy (TG-IR), the transformation of xylan, cellulose, and lignin contents was observed under various pHs and DIC concentrations. PMID- 17410784 TI - C-1s NEXAFS spectroscopy reveals chemical fractionation of humic acid by cation induced coagulation. AB - The influence of cation-induced coagulation on the chemical composition of dissolved and coagulated fractions of humic acid was investigated in batch coagulation experiments for additions of aluminum at pH 4 and 5, iron at pH 4, and calcium and lead at pH 6. The partitioning of organic carbon and metals was determined by analyzing total organic carbon and total metal contents of the dissolved phase. Both the dissolved and the coagulated humic acid fractions were characterized using synchrotron scanning transmission X-ray microscopy (STXM) and C-1s near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. Intensities of pi* transitions of carboxyl carbon and sigma* transitions of alkyl, O-alkyl, and carboxyl carbon decreased with increasing metal concentration for the dissolved humic acid fractions. This decrease was accompanied by an increase of the respective intensities in the coagulated fraction as shown for lead. Intensities of aromatic and phenolic carbon were affected to a larger extent only by aluminum and iron additions. The changes observed in the C-1s NEXAFS spectra coincided with an increasing removal of organic carbon from the dissolved phase with increasing total metal concentrations. We conclude that humic acid was chemically fractionated by cation-induced coagulation, which preferentially removed functional groups involved in metal-cation binding from solution. PMID- 17410785 TI - Hydrogarnet: a host phase for Cr(VI) in chromite ore processing residue (COPR) and other high pH wastes. AB - For understanding both the environmental behavior and developing remediation treatments for chromium ore processing residue (COPR) it is important to identify all the potentially soluble sources of Cr(VI). Hydrogarnet has been identified as a major phase in COPR and it has been previously speculated that it has a capacity to host Cr(VI). Here we provide direct evidence of this capacity by demonstrating the incorporation of Cr(VI) into laboratory synthesized hydrogarnet. Electron microscopy and energy dispersive X-ray microanalysis show incorporation of approximately 17000-22000 mg Cr(VI) kg(-1) hydrogarnet. X-ray powder diffraction data show that peak intensities are altered by chromium substitution and that chromium substituted hydrogarnets have a smaller unit cell than the pure Ca-Al end member. This is consistent with substitution of hydroxyl tetrahedra by smaller chromate tetrahedra. Electron energy loss spectroscopy confirms the tetrahedral coordination and hexavalent oxidation state of chromium in the hydrogarnets. The maximum amount of hexavalent chromium that can be introduced synthetically corresponds to a replacement of about one out of every eight hydroxyl tetrahedral per unit cell by a CrO4(2-) tetrahedra and tallies closely with the amount of chromium measured in hydrogarnets from COPR. Chromium bearing hydrogarnet is the most abundant crystalline phase in millions of tons of COPR contaminating land around Glasgow, Scotland, and was recently identified in COPR from sites in North America. Calculations based on its abundance and its Cr(VI) content indicate that hydrogarnet can host as much as 50% of the Cr(VI) found in some COPR samples. PMID- 17410786 TI - Sorption of tetracycline and chlortetracycline on K- and Ca-saturated soil clays, humic substances, and clay-humic complexes. AB - Tetracycline (TC) and chlortetracycline (CTC) are used extensively for growth promotion and therapeutic purposes in livestock production. The sorption of TC and CTC on clays, humic substances (HS), and clay-humic complexes (clay-HC) derived from two agricultural soils was quantified using dilute CaCl2 (Ca) and KCI (K) as background solutions. In all systems, the soil components sorbed > 96% of added tetracyclines. Strongest sorption was observed for clays, followed by HS, and then clay-HC. Greater sorption by the Ca systems than the K systems and decreased sorption with increasing pH suggests that cation bridging and cation exchange contribute to sorption. X-ray diffraction analysis showed that TC and CTC were sorbed in the interlayers of smectites and that the presence of HS reduced interlayer sorption of tetracyclines by smectites in clay-HC. The results indicate that tetracyclines are dominantly sorbed on soil clays and that HS in clay-HC either mask sorption sites on clay surfaces or inhibit interlayer diffusion of tetracyclines. PMID- 17410787 TI - Sorption and inhibited dehydrohalogenation of 2,2-dichloropropane in micropores of dealuminated Y zeolites. AB - Contaminant transformation rates in the subsurface can be slowed by sorption onto geosorbents. We evaluated the effect of micropore sorption on contaminant transformation in a model system consisting of dealuminated Y zeolites and 2,2 dichloropropane (2,2-DCP). 2,2-DCP dehydrochlorinates in water to 2-chloropropene (2-CP) at a rate of 2.93 x 10(-4) min(-1) at 24 degrees C. The Y zeolites used range from hydrophilic (CBV-300) to hydrophobic (CBV-720 and CBV-780). Wet zeolite samples were loaded with 2,2-DCP at 24 degrees C and reacted at 50 degrees C for 10 h. Results show that the hydrophobic zeolites (CBV-720 and CBV 780) sorbed nearly 900 times more 2,2-DCP than the hydrophilic CBV-300 under wet conditions. 2,2-DCP transformed less when sorbed in micropores of CBV-720 (6.3%) and CBV-780 (5.0%) than in micropores of CBV-300 (81.5%), and significantly less than in water (> 99.85%). Inhibition in hydrophobic micropores is interpreted as lack of water solvating the transition state of 2,2-DCP dehydrohalogenation and the H+ and Cl- formed. Near the micropore openings, the transformation was relatively fast, consistent with greater abundance of water associated with the hydrophilic edge sites. These results indicate that in the subsurface the half lives of reactive organic contaminants may be longer than predicted from bulk water data. PMID- 17410788 TI - Zn incorporation in hydroxy-Al- and keggin Al13-intercalated montmorillonite: a powder and polarized EXAFS study. AB - The sorption mechanism of Zn on gibbsite and mont-morillonite exchanged with Al3+ (Al-mont) or Keggin Al13 polymer (Al13-mont) was probed by powder and polarized EXAFS spectroscopy as a function of pH (5.85-7), reaction time (1-65 days), and sorbate to sorbent ratio (50-Zn is octahedrally coordinated to oxygens at approximately 2.08(2) A, and surrounded in-plane by six Al atoms at 3.02-3.06(2) A, and another six at 6 A. No out-of plane Si neighbors are detected. These results are interpreted as Zn incorporation in vacant octahedral sites of gibbsite-like layers at the basal and/or interlayer surface of montmorillonite particles. Zinc sorbed on the edges of gibbsite layers would give a split first oxygen shell with bond distances of 2.00(2) and 2.16(3) A, and 2.1(8) nearest Al at 3.02 A with no second-nearest Al, none of which were observed in Al-mont. The binding environment of Zn on Al13-mont after 1 day is similar to that on the edges of gibbsite, and is interpreted as Zn complexation at the surface of Al polymers. After 28 days, the Zn environment resembles that of Zn-sorbed Al-mont, indicating the progressive buildup of Zn-containing gibbsite-like layers parallel to montmorillonite layers. The results of this work clarify the incorporation mechanism of Zn in hydroxy-Al interlayered phyllosilicate and provide insight on the formation mechanism of this common Zn species in soil. PMID- 17410789 TI - Noble gas excess air applied to distinguish groundwater recharge conditions. AB - The application of geochemical tracers in groundwater studies can provide valuable insights into the rates and sources of groundwater recharge, residence times, and flow dynamics that are of significant value in the management of this important natural resource. This paper demonstrates the application of noble gas excess air to distinguish groundwater bodies with different recharge histories in a layered sandstone aquifer system in the east of England. The sampled groundwaters are all supersaturated with respect to neon, indicating the presence of excess air. The lowest excess air concentrations occur where the aquifer is unconfined (deltaNe, the proportion of neon in excess of saturation, = 12-26%) and recharge occurs directly to the outcrop. Groundwater in the confined part of the aquifer can be divided into two hydrochemical types based upon the dissolved ion chemistry: Type 1 groundwaters contain more excess air (deltaNe = 115-120%) than Type 2(deltaNe = 22-62%). The difference in excess air concentrations confirms that groundwater enters the confined aquifer along two discrete pathways. Furthermore, excess neon concentrations predicted from the magnitude of annual water table fluctuation observed in the different recharge areas are in good agreement with those measured in the corresponding groundwaters. We therefore recommend that excess air may be usefully employed as a direct indicator of the volume of long-term net annual groundwater recharge. PMID- 17410790 TI - Measuring air-water interfacial areas with X-ray microtomography and interfacial partitioning tracer tests. AB - Air-water interfacial areas as a function of water saturation were measured for a sandy, natural porous medium using two methods, aqueous-phase interfacial partitioning tracer tests and synchrotron X-ray microtomography. In addition, interfacial areas measured in a prior study with the gas-phase interfacial partitioning tracer-test method for the same porous medium were included for comparison. For all three methods, total air-water interfacial areas increased with decreasing water saturation. The interfacial areas measured with the tracer test methods were generally larger than those obtained from microtomography, and the disparity increased as water saturation decreased. The interfacial areas measured by microtomography extrapolated to a value (147 cm(-1)) very similar to the specific solid surface area (151 cm(-1)) calculated using the smooth-sphere assumption, indicating that the method does not characterize the area associated with microscopic surface heterogeneity (surface roughness, microporosity). This is consistent with the method resolution of approximately 12 microm. In contrast, the interfacial areas measured with the gas-phase tracer tests approached the N2/BET measured specific solid surface area (56000 cm(-1)), indicating that this method does characterize the interfacial area associated with microscopic surface heterogeneity. The largest interfacial area measured with the aqueous-phase tracer tests was 224 cm(-1), while the extrapolated maximum interfacial area was approximately 1100 cm(-1). Both of these values are larger than the smooth-sphere specific solid surface area but much smaller than the N2/BET specific solid surface area, which suggests that the method measures a limited portion of the interfacial area associated with microscopic surface heterogeneity. All three methods provide measures of total (capillary + film) interfacial area, a primary difference being that the film-associated area is a smooth-surface equivalent for the microtomography method. An advantage of the microtomography method is the ability to determine explicitly both total and capillary-associated interfacial areas, which is problematic for the tracer-test methods. PMID- 17410791 TI - Size distribution of trace organic species emitted from heavy-duty diesel vehicles. AB - Size distributions of particulate hopanes, steranes, and polycyclic aromatic hydrocarbons (PAHs) were measured in the exhaust from four heavy-duty diesel vehicles (HDDVs) operated under idle, creep, transient, and two high-speed driving modes. Particulate matter was collected using a chassis dynamometer and a dilution sampling system equipped with cascade impactors and filter samplers. Samples were extracted using organic solvents and analyzed using gas chromatography-mass spectrometry. Size distributions of hopanes and steranes were functions of engine load conditions and vehicle technology. Hopanes and steranes peaked in size ranges larger than 0.18 microm aerodynamic particle diameter under light load conditions and less than 0.10 microm aerodynamic particle diameter under heavier load conditions. The eight hopane size distributions emitted from newertechnology (> 1998) vehicles were unimodal while the four hopane size distributions emitted from older technology vehicles (< 1992) were bimodal. Similar trends between older and newer vehicles were not observed for sterane size distributions. The PAH composition emitted from HDDVs was a function of driving cycle and vehicle technology. Light driving cycles produced quantifiable emissions of 3, 4, 5, and 6 ring PAHs (including coronene). Heavier driving cycles produced only the 3 and 4 ring PAHs in quantifiable amounts. PM1.8 and PM0.1 source profiles constructed using the relative abundance of hopanes and steranes to total organic carbon were functions of vehicle load condition. Increasing load reduced the relative abundance of motor oil tracers in the PM1.8 size fraction and increased the abundance of these tracers in the PM0.1 size fraction. The relative abundances of PAHs in the PM0.1 and PM1.8 size fractions emitted from the oldest vehicle tested (1985 HDDV) were significantly higher than for any other vehicle tested. PMID- 17410792 TI - Estimation of dry deposition of atmospheric mercury in Nevada by direct and indirect methods. AB - Atmospheric models and limited measurements indicate that dry deposition of atmospheric mercury is an important process by which mercury is input to ecosystems. To begin to fill the measurement data gap, multiple methods were used simultaneously during seasonal campaigns conducted in 2005 and 2006 to estimate dry deposition of atmospheric mercury at two Mercury Deposition Network (MDN) sites in rural Nevada and in Reno, Nevada. Gaseous elemental mercury (Hg0), reactive gaseous mercury (RGM), and particulate-bound mercury (Hgp) concentrations were measured using Tekran 2537A/1130/ 1135 systems. These speciated measurements were combined with on-site meteorological measurements to estimate depositional fluxes of RGM and Hgp using dry deposition models. Modeled fluxes were compared with more direct measurements obtained using polysulfone cation-exchange membranes and foliar surfaces. Dynamic flux chambers were used to measure soil mercury exchange. RGM concentrations were higher during warmer months at all sites, leading to seasonal variation in the modeled importance of RGM as a component of total depositional load. The ratio of dry to wet deposition was between 10 and 90%, and varied with season and with the methods used for dry deposition approximations. This work illustrates the variability of mercury dry deposition with location and time and highlights the need for direct dry deposition measurements. PMID- 17410793 TI - Kinetics and mechanisms of radiolytic degradation of nitrobenzene in aqueous solutions. AB - Steady-state radiolysis experiments were performed to gain insight into the kinetics and mechanisms of nitrobenzene (NB) degradation in aqueous solutions. The degradation of NB under gamma-ray irradiation followed pseudo-first-order kinetics, and the pseudo first-order rate constant and the initial G value of NB decomposition were functionally related to both the initial NB concentration and the irradiation dose rate. Under oxidative conditions, complete mineralization of NB was achieved, whereas no total organic carbon reduction was observed under reductive conditions. The radiolytic products of NB under various conditions were identified using FTIR and GC-MS analyses. The mechanisms behind the radiolytic degradation of NB under both oxidative and reductive conditions were proposed schematically in light of the degradation products observed. In addition, calculations based on ab initio molecular orbital and density functional theory provided support for the proposed mechanisms and the preferred pathways among all the possible reactions. PMID- 17410794 TI - Continuous catalytic hydrogenation of polyaromatic hydrocarbon compounds in hydrogen-supercritical carbon dioxide. AB - A continuous hydrogenation device was evaluated for the detoxification of selected tri-, tetra-, or pentacyclic polyaromatic hydrocarbon (PAH) compounds {anthracene, phenanthrene, chrysene, and benzo[a]pyrene (B[a]P)} by hydrogenation. A substrate stream in hexane, 0.05-1.0% (w/v), was mixed with hydrogen-carbon dioxide (H2-CO2, 5-30% v/v) and delivered to a heated reactor column (25 cm x 1 cm) containing palladium supported on gamma alumina (Pd0/gamma Al2O3) that was terminated with a capillary restrictor. The flow rate from the reactor, approximately 800 mL min(-1) decompressed gas, corresponded to 4 mL min( 1) fluid under the operating conditions of the trials. Reaction products were recovered by passing the reactor effluent through hexane. At 90 degrees C, the anthracene or phenanthrene substrate was hydrogenated only partially to octahydro and dodecahydro species and contained only a minor quantity of totally hydrogenated products. For substrates with increasing numbers of fused aromatic rings, the hydrogenation efficiency was decreased further. However, at an increasing temperature (90-150 degrees C) and increasing mobile phase flow rate (20.68 MPa corresponding to 2100 mL min(-1) decompressed gas), B[a]P and chrysene were hydrogenated, virtuallytotally, to their corresponding perhydro analogues (eicosahydrobenzo[a]pyrenes and octadecahydrochrysenes), respectively. That this approach might be useful for decontaminating soil extracts was supported by companion in vitro trials in which the substrate and products were assayed for mutagenic activity with five bacterial strains that are auxotrophic for histidine (Salmonella typhimurium TA98, TA100, TA1535, and TA1537) or tryptophan (Escherichia coliWP2 uvrA), using the bacterial reverse mutation assay (modified Ames test). Generally, substantial increases in revertant colony counts were not observed with any of the strains following exposure to the hydrogenation products in the absence or presence of the 10 or 30% S9 mix, which is consistent with the loss of mutagenic activity from these hydrogenation products. PMID- 17410795 TI - Precipitates on granular iron in solutions containing calcium carbonate with trichloroethene and hexavalent chromium. AB - Mineralogical examination, using scanning electron microscopy (SEM), X-ray diffractometry (XRD), and optical microscopy, was conducted on the Fe0-bearing reactive materials derived from long-term column experiments undertaken to assess the treatment capacity of Fe0 under different geochemical conditions. The columns received either deionized water or solutions of differing dissolved calcium carbonate concentrations, together either with trichloroethene (TCE) or hexavalent chromium (Cr(VI)). The major reaction product in the columns receiving deionized water was magnetite-maghemite, and for the columns receiving dissolved calcium carbonate, the main products were iron hydroxy carbonate and aragonite. Replacement of Fe0 by reaction products occurred mainly at the edges of the Fe0 particles, and penetrative replacement was focused along cracks and along and around graphitic inclusions. Fibrous or flake-shaped iron hydroxy carbonate mostly replaced the edges of the Fe0 particles. Aragonite had needle-shaped morphology, and some occurred as clusters of crystals. Aragonite was deposited on iron hydroxy carbonate, thus providing at least a partial armoring effect. The mineral was also observed to cement groups of Fe0 particles into compact aggregates. The Cr was present mostly as Cr(III) in Cr(III)-Fe(III) (oxy)hydroxides and in trace amounts in iron hydroxy carbonate. PMID- 17410796 TI - Recycle technology for recovering resources and products from waste printed circuit boards. AB - The printed circuit board (PCB) contains nearly 28% metals that are abundant non ferrous metals such as Cu, Al, Sn, etc. The purity of precious metals in PCBs is more than 10 times higher than that of rich-content minerals. Therefore, recycling of PCBs is an important subject not only from the treatment of waste but also from the recovery of valuable materials. Chemical and mechanical methods are two traditional recycling processes for waste PCBs. However, the prospect of chemical methods will be limited since the emission of toxic liquid or gas brings secondary pollution to the environment during the process. Mechanical processes, such as shape separation, jigging, density-based separation, and electrostatic separation have been widely utilized in the recycling industry. But, recycling of waste PCBs is only beginning. In this study, a total of 400 kg of waste PCBs was processed by a recycle technology without negative impact to the environment. The technology contained mechanical two-step crushing, corona electrostatic separating, and recovery. The results indicated that (i) two-step crushing was an effect process to strip metals from base plates completely; (ii) the size of particles between 0.6 and 1.2 mm was suitable for corona electrostatic separating during industrial application; and (iii) the nonmetal of waste PCBs attained 80% weight of a kind of nonmetallic plate that expanded the applying prospect of waste nonmetallic materials. PMID- 17410797 TI - Minimizing dioxin emissions from integrated MSW thermal treatment. AB - The combustion of wastes has very significant benefits in reducing the volume of waste materials and producing energy. However, combustion processes produce emissions, which must be below the Best Practical Means (BPM) specified legislative limits. Several wastes, such as tires and meat meal, have been successfully combusted in cement kilns, up to 20% w/w, while retaining emission standards well below legislative limits. In the case of municipal solid waste (MSW) the introduction of large amounts of MSW into cement kilns is not practical because the additional kiln volume required is too great, the large amounts of ash generated will affect the cement clinker quality, and it would be difficult to sustain the required very high clinkering temperature of 1500 degrees C with large quantities of low calorific value MSW. A completely novel process, termed the Co-Co process, has been developed, integrating MSW combustion in a synergistic fashion with the cement production. This process is based on combining the cement "front-end" calcination reaction and incorporating it with a high temperature, at 1200 degrees C, combustion process, providing a giant acid gas scrubber. A pilot plant was designed, constructed, and operated to demonstrate the benefits of the Co-Co process. The pilot plant achieved emissions minimization: dioxins were typically 0.5-1% of the European BPM limits, HCl, SOx, NOx, and particulates were 15, 10, 20, and 25% of BPM limits, respectively. Heavy metals were typically below 25% of BPM limit values. PMID- 17410798 TI - Effect of flux (transmembrane pressure) and membrane properties on fouling and rejection of reverse osmosis and nanofiltration membranes treating perfluorooctane sulfonate containing wastewater. AB - Perfluorooctane sulfonate (PFOS) is an emergent contaminant of substantial environmental concerns. In this study, reverse osmosis (RO) and nanofiltration (NF) membranes were used to remove this toxic and persistent compound from PFOS containing wastewater. Five RO membranes and three NF membranes were tested at a feed concentration of 10 ppm PFOS over 4 days, and the PFOS rejection and permeate flux performances were systematically investigated. PFOS rejection was well correlated to sodium chloride rejection. The rejection efficiencies for the RO membranes were > 99%, and those for the NF membranes ranged from 90-99%. Improvement in PFOS rejection, together with mild flux reduction (< 16%), was observed at longer filtration time. Such shifts in rejection and flux performance were probably due to the increased PFOS accumulation at longer duration, as shown by X-ray photoelectron spectroscopy and liquid chromatograph and tandem mass spectrometry results. A fraction of PFOS molecules might be entrapped in the polyamide layer of the composite membranes, which hindered the further passage of both water and other PFOS molecules. In a similar fashion, PFOS rejection and fouling were enhanced for greater initial flux and/or applied pressure, where PFOS accumulation was promoted probably due to increased hydrodynamic permeate drag. Flux reduction was also shown to correlate to membrane roughness, with the rougher membranes tend to experience more flux reduction than the smoother ones. PMID- 17410800 TI - Adsorption of humic acid onto nanoscale zerovalent iron and its effect on arsenic removal. AB - Batch experiments were performed to investigate the feasibility of humic acid (HA) removal by synthetic nanoscale zerovalent iron (NZVI) and its interaction with As(III) and As(V), the most poisonous and abundant of groundwater pollutants. High-resolution transmission electron microscopy (HR-TEM) and X-ray diffraction (XRD) were used to characterize the particle size, surface morphology of the pristine NZVI and HA-treated NZVI (NZVI-HA), and the zero valence state of the pristine NZVI. It was determined that HA was completely removed by NZVI (0.3 g/L) within a few minutes, at a wide range of initial pH values (approximately 3.0-12.0). Fourier transform infrared (FTIR) and laser light scattering (zeta potential measurement) studies confirmed that NZVI-HA forms inner-sphere surface complexation at different initial pH conditions. The effects of competing anions showed that there was complete removal of HA in the presence of 10 mM NO(-3) and SO4(2-) whereas HA removal was observed 0%, 18% and 22% in presence of 10 mM H2PO4(2-), HCO(3-) and H4SiO4(0), respectively. However, the presence of 2 mM CA2+ and Mg2+ enhanced HA removal from 17 mg g(-1) to 76 mg g(-1) and 55 mg g( 1), respectively. Long-term time-resolved studies of XRD and field emission scanning electron microscopy (FE-SEM) with energy-dispersive X-ray (EDX) revealed the formation of various types of new iron oxides (magnetite, maghemite, and lepidocrocites) during the continuous reaction of HA in the presence of water and NZVI at 1, 30, 60, and 90 days. In addition, the surface-area-normalized rate constant (ksa) of adsorption of As(III) and As(V) onto NZVI was reduced in the presence of HA (20 mg L(-1)), from 100% to 43% and 68%, respectively. Our results show the potential use of NZVI in removing HA and its possible effects on arsenic removal during the application of NZVI in groundwater remediation. PMID- 17410799 TI - Impact of ethanol on the natural attenuation of MTBE in a normally sulfate reducing aquifer. AB - Side-by-side experiments were conducted in an aquifer contaminated with methyl tert-butyl ether (MTBE) at a former fuel station to evaluate the effect of ethanol release on the fate of pre-existing MTBE contamination. On one side, for approximately 9 months we injected groundwater amended with 1-3 mg/L benzene, toluene, and o-xylene (BToX). On the other side, we injected the same, adding approximately 500 mg/L ethanol. The fates of BToX in both sides ("lanes") were addressed in a prior publication. No MTBE transformation was observed in the "No Ethanol Lane." In the "With Ethanol Lane", MTBE was transformed to tert-butyl alcohol (TBA) underthe methanogenic and/or acetogenic conditions induced by the in situ biodegradation of the ethanol downgradient of the injection wells. The lag time before onset of this transformation was less than 2 months and the pseudo-first-order reaction rate estimated after 7-8 months was 0.046 d(-1). Our results imply that rapid subsurface transformation of MTBE to TBA may be expected in situations where strongly anaerobic conditions are sustained and fluxes of requisite nutrients and electron donors allow development of an active acetogenic/methanogenic zone beyond the reach of inhibitory effects such as those caused by high concentrations of ethanol. PMID- 17410801 TI - Evaluation of the intrinsic photocatalytic oxidation kinetics of indoor air pollutants. AB - This paper presents a methodology for the evaluation of the intrinsic photocatalytic oxidation (PCO) kinetics of indoor air pollutants. It combines computational fluid dynamics (CFD) modeling of the fluid flow in the reactor with radiation field modeling and photocatalytic reaction kinetics to yield a rigorous model of a flat-plate, single-pass, flow-through photocatalytic reactor for indoor air purification. This method was applied to model the PCO of trichloroethylene (TCE) in humidified air and to derive kinetic parameters directly from kinetic data in an integral flow reactor. Steady-state PCO experiments of TCE over irradiated TiO2 (Degussa P25) thin films immobilized on glass supports were carried out at different radiation intensities, flow rates, and inlet substrate concentrations. The oxidation rate of TCE was found to be first-order on the incident photon flux and to follow a Langmuir-Hinshelwood type reaction kinetics rate law. Mass transfer resistances were observed at Reynolds numbers less than 46. Apparent quantum yields were found to be up to 0.97 mol Einstein(-1). A comparison of the model prediction with the experimental results in an integral reactor yielded pollutant-specific kinetic rate parameters which were independent of reactor geometry, radiation field, and fluid-dynamics. The kinetic parameters would,therefore, be more universally applicable to the design and scale-up of photocatalytic reactors for indoor air purification. PMID- 17410803 TI - Efficient heterogeneous catalytic reduction of perchlorate in water. AB - A new heterogeneous catalyst that promotes the reduction by hydrogen of perchlorate ion in water under mild conditions has been developed. The catalyst is prepared by adsorption of a rhenium(VII) precursor (either ammonium perrhenate or methylrhenium trioxide) onto carbon powder containing 5% palladium by weight. Under standard batch conditions of room temperature, 1 bar of hydrogen, and 200 ppm perchlorate (as HClO4), reduction proceeded to less than 1 ppm in as little as 5 h. Extended reaction times led to residual perchlorate at low parts per billion levels. Chloride was the only observed product, with good material balance. Catalytic materials ranging from 3% to 13% Re showed (pseudo) first order rates linearly dependent on the Re content. Representative normalized rate constants for catalysts with 5-9% Re were in the range 0.1-0.3 L h(-1) (g of cat.)(-1). Inhibition by chloride was not significant, with little change in perchlorate reduction rate in the presence of excess chloride to 1000 ppm. However, optimal activity occurred in acidic solutions (pH ca. 3), and both the rate and extent of reaction decreased at higher values of pH. In its current form the catalyst might be best applied to destroy perchlorate in the acidic regeneration stream from selective ion exchange columns. PMID- 17410802 TI - Variations in 13C/12C and D/H enrichment factors of aerobic bacterial fuel oxygenate degradation. AB - Reliable compound-specific isotope enrichment factors are needed for a quantitative assessment of in situ biodegradation in contaminated groundwater. To obtain information on the variability on carbon and hydrogen enrichment factors (epsilonC, epsilonH) the isotope fractionation of methyl tertiary (tert-) butyl ether (MTBE) and ethyl tert-butyl ether (ETBE) upon aerobic degradation was studied with different bacterial isolates. Methylibium sp. R8 showed a carbon and hydrogen isotope enrichment upon MTBE degradation of -2.4 +/- 0.1 and -42 +/- 4 per thousand, respectively, which is in the range of previous studies with pure cultures (Methylibium petroleiphilum PM1) as well as mixed consortia. In contrast, epsilonC of the beta/-proteobacterium L108 (-0.48 +/- 0.05 per thousand) and Rhodococcus ruber IFP 2001 (-0.28 +/- 0.06 per thousand) was much lower and hydrogen isotope fractionation was negligible (epsilonH < or = -0.2 per thousand). The varying isotope fractionation pattern indicates that MTBE is degraded by different mechanisms by the strains R8 and PM1 compared to L108 and IFP 2001. The carbon and hydrogen isotope fractionation of ETBE by L108 (epsilonC = -0.68 +/- 0.06 per thousand and epsilonH = -14 +/- 2 per thousand) and IFP 2001 (epsilonC = -0.8 +/- 0.1 per thousand and epsilonH = -11 +/- 4 per thousand) was very similar and seemed slightly higher than the fractionation observed upon MTBE degradation by the same strains. The low carbon and hydrogen enrichment factors observed during MTBE and ETBE degradation by L108 and IFP 2001 suggest a hydrolysis-like reaction type of the ether bond cleavage compared to oxidation of the alkyl group as suggested for the strains PM1 and R8. The variability of carbon and hydrogen enrichment factors should be taken into account when interpreting isotope pattern of fuel oxygenates with respect to biodegradation in contamination plumes. PMID- 17410804 TI - Quantification and modeling of the elimination behavior of ecologically problematic wastewater micropollutants by adsorption on powdered and granulated activated carbon. AB - The adsorption on powdered activated carbon (PAC) of ecotoxic or potentially ecotoxic micropollutants (ten pharmaceuticals, four X-ray contrast media, and eight industrial chemicals) present in a biologically treated municipal wastewater is studied. All but the X-ray contrast media are eliminated with high efficiency at an economically feasible PAC dosage of 10 mg/L. Based on the experimental data, the competition between the background organic matter and the micropollutant for the active sites of the adsorbent is modeled with the help of the adsorption and tracer analysis supported by the Ideal Adsorption Solution Theory. With granulated activated carbon, adsorption isotherms are determined by spiking. Based on these experimental data and modeled parameters, a lay-out of fixed-bed adsorbers may be simulated. PMID- 17410805 TI - Oxidation of N-nitrosodimethylamine (NDMA) precursors with ozone and chlorine dioxide: kinetics and effect on NDMA formation potential. AB - The oxidation of N-nitrosodimethylamine (NDMA) precursors chlorine dioxide (ClO2). Second-order rate constants for the reactions of model NDMA precursors (dimethylamine (DMA) and 7 tertiary amines) with ozone (kapp at pH 7 = 2.4 x 10( 1) to 2.3 x 10(9) M(-1) s(-1)), ClO2 (kapp at pH 7 = 6.7 x 10(-3) to 3.0 x 10(7) M(-1) s(-1)), and hydroxyl radical (*OH) (kapp at pH 7 = 6.2 x 10(7) to 1.4 x 10(10) M(-1) s(-1)) were determined, which showed that the selected NDMA precursors, with the exception of dimethylformamide (DMFA) can be completely transformed via their direct reaction with ozone. During ozonation, DMFA may be partially transformed through oxidation by the secondary oxidant *OH. ClO2 was also shown to effectively transform most of the precursors, with the exceptions of DMA and DMFA. In the second part of the study, the NDMA formation potentials (NDMA-FP) in synthetic and natural waters were measured with and without pre oxidation with ozone and ClO2. A significant reduction in the NDMA-FPs was observed after complete transformation of the model NDMA precursors. Ozonation generally led to more effective reduction of the NDMA-FP than ClO2. For most of the precursors, the formation of DMA could account for the NDMA-FPs remaining after complete transformation of the model NDMA precursors. In contrast, dimethylethanolamine and dimethyldithiocarbamate yielded other NDMA precursors (not DMA) as their oxidation products. Pre-oxidation by ozone and ClO2 of several natural waters showed behavior similar to that of the oxidation of model NDMA precursors with a reduction of the NDMA-FP by 32-94% for various natural water sources. PMID- 17410806 TI - In vitro cytotoxicitiy of silica nanoparticles at high concentrations strongly depends on the metabolic activity type of the cell line. AB - Amorphous silica is increasingly used in diagnostic and biomedical research because of its ease of production and relatively low cost. It is generally regarded as safe and has been approved for use as a food or animal feed ingredient. Recent literature reveals that amorphous silica may present toxicity concerns at high doses. In anticipation of potential human exposure to silica, it is advisable to examine its toxicity to cells of different organs. Consequently, we investigated the response of several normal fibroblast and tumor cells to varying doses of amorphous silica or composite nanoparticles of silica and chitosan. A cell proliferation assay indicates that silica nanoparticles are nontoxic at low dosages but that cell viability decreases at high dosages. A lactate dehydrogenase (LDH) assay indicates that high dosages of silica induce cell membrane damage. Both assays reveal that fibroblast cells with long doubling times are more susceptible to injury induced by silica exposure than tumor cells with short doubling times. In contrast, silica-chitosan composite nanoparticles induce less inhibition in cell proliferation and less membrane damage. This study suggests that the cytotoxicity of silica to human cells depends strongly on their metabolic activities but that it could be significantly reduced by synthesizing silica with chitosan. PMID- 17410807 TI - Anemia and cancer in older persons. AB - Although the overall prevalence for many cancers is declining, cancer still remains a diagnosis more common in the elderly than in younger individuals. As the population ages, the proportion of patients with cancer who are elderly is expected to increase dramatically. Anemia occurs more often in older individuals for a variety of reasons, and its prevalence in elderly patients with cancer is significantly increasing. Although information on anemia in the elderly is limited, data on cancer patients of all ages have shown that the presence of anemia is associated with poorer prognosis and functional status. The impact of anemia on performance status in patients with cancer is matched by results from a large number of studies indicating negative effects of anemia on a wide variety of performance measures. Despite an apparent reluctance to undertake aggressive therapy in elderly patients with cancer, physicians are using a wider range of treatments with increasing frequency in this older population. Optimizing treatment outcomes in elderly patients with cancer depends on careful determination of performance status as well as the potential of therapies to treat anemia in these patients. PMID- 17410808 TI - An overlooked problem: anemia in the elderly patient with cancer. PMID- 17410809 TI - Aging, cancer, and anemia: intertwined and resolvable. PMID- 17410810 TI - Symptom control trials: a 20-year experience. AB - Symptoms related to cancer and/or cancer therapy are a prominent consideration in cancer patients; multiple options have been proposed to alleviate these problems. The US National Cancer Institute (NCI) Community Clinical Oncology Program (CCOP) has mandated that CCOP participant institutions be involved with cancer control trials in addition to cancer treatment trials. Through such efforts, scientifically sound symptom control clinical trials are being conducted to determine the effectiveness of promising agents. In this article, the authors provide an update of clinical trial outcomes regarding 13 symptom and symptom complexes related to cancer and/or cancer therapy, emphasizing that publication of both positive and negative study results is important to separate what works from what does not. PMID- 17410811 TI - The right stuff to advance the science of comfort. PMID- 17410812 TI - Use of warfarin in the patient with cancer. PMID- 17410813 TI - Oncologist-patient discussion of adjuvant hormonal therapy in breast cancer: results of a linguistic study focusing on adherence and persistence to therapy. AB - Although studies have proven the benefit of 5+ years of adjuvant hormonal therapy (AHT) for breast cancer, data show adherence and persistence with therapy are suboptimal. This observational linguistic study analyzed communication between breast cancer patients and their oncologists to determine how adherence was addressed and to identify areas where communication could be improved. Community based oncologists were recruited by letter to participate. Researchers visited oncologists (n = 14) to record patient-oncologist interactions and conduct separate post-visit interviews. Comprehensive linguistic analyses of visits between 28 postmenopausal, early-stage breast cancer patients on or initiating hormonal therapy and their oncologists were conducted to determine the nature of discussions of adherence and persistence to therapy. Oncologist-patient discussions about AHT were generally good but did not address potential difficulties of remaining adherent with long-term therapy. Discussions of persistence were usually monologues addressing the current state of "study data" and were not linked to the patient, the importance of persistence, or how the study data related to her situation. Because the patient's cancer was framed as being "in the past," discussions resembled those of chronic management in preventive medicine. This more ad-hoc approach to adherence and persistence is a potential stumbling block for motivating patients to stay on hormonal therapy. Additionally, the oncologists participating in this study recognized that adherence to therapy is a problem but did not feel "their patients" fell into this pattern. In this office-based evaluation, minimal nurse interactions were observed, which increases the importance of oncologist-patient communication. The authors recommend that oncologists leverage the existing good communication with their patients by increasing the amount and quality of discussions around the importance of adherence and persistence to AHT. PMID- 17410814 TI - All patient refined-diagnostic related group and case mix index in acute care palliative medicine. AB - The All Patient Refined-Diagnostic Related Group (APR-DRG) is a modification of the traditional DRG that adds four classes of illness severity and four classes of mortality risk. The APR-DRG is a more accurate assessment of the complexity of care. When individuals with advanced illness are admitted to an acute inpatient palliative medicine unit, there may be a perception that they receive less intense acute care. Most of these patients, however, are multisymptomatic, have several comorbidities, and are older. For all patients admitted to the unit, a guide was followed by staff physicians to document clinical information that included the site(s) of malignancy, site(s) of metastases, disease complications, disease-related symptoms, and comorbidities. We then prospectively compared DRGs, APR-DRGs, and case mix index (CMI) from January 1-June 30, 2003, and February 1 July 31,2004, before and after the use of the guide. The overall mean severity of illness (ASOI) increased by 25% (P < 0.05). The mean CMI increased by 12% (P < 0.05). The average length of stay over the same period increased slightly from 8.97 to 9.56 days. Systematic documentation of clinical findings using a specific tool for patients admitted to an acute inpatient palliative medicine unit based on APR-DRG classifications captured a higher severity of illness and may better reflect resource utilization. PMID- 17410815 TI - Hanging up the scalpel: knowing when to say "when". PMID- 17410816 TI - What makes you angry makes you motivated. PMID- 17410817 TI - The $22 billion gold rush. PMID- 17410818 TI - Litigation plays protective role--and is part of the problem. PMID- 17410819 TI - Missouri's Constitutional Amendment No. 2 & stem cell research: some ethical and political considerations. PMID- 17410820 TI - The Patient Safety and Quality Improvement Act of 2005--The federal law and its implications for Mssouri. AB - The Patient Safety and Quality Improvement Act of 2005 establishes a network of federally certified Patient Safety Organizations (PSO). PSOs will establish voluntary, confidential data systems and forums for providers to learn and improve patient safety. The Act provides protections, not available in Missouri, from discovery of data reported to a PSO. Providers should begin assessing patient safety activities and benefits of PSO participation. The Missouri Center for Patient Safety plans to become a statewide PSO. PMID- 17410821 TI - Overseas medicine--part II. Military medicine in Iraq: equal care for all. PMID- 17410822 TI - What is the scope of physician-patient relationship? PMID- 17410823 TI - Geriatrics: the specialty of the future. PMID- 17410824 TI - Comprehensive geriatric assessment. AB - This paper presents the essentials of the comprehensive assessment of the geriatric patient. PMID- 17410825 TI - Assessment of appetite and weight loss syndromes in nursing home residents. AB - This article presents an appropriate definition of the construct of appetite and outlines an evidence based approach to appetite assessment and weight loss prevention in the nursing home. The Simplified Nutritional Appetite Questionnaire, a recently developed and validated appetite assessment tool that predicts weight loss in older and young nursing home residents and community dwelling adults is discussed. PMID- 17410826 TI - Prevention and management of pressure ulcers. AB - Pressure ulcers represent complex wounds that are difficult to prevent or manage. Guidelines for prevention include identifying patients at risk, reducing the effect of pressure, friction, shear forces, and assessing co-morbidities such as nutritional status. Management should follow eight treatment strategies including accurately assessing the ulcer, relieving pressure, assessing pain and nutritional status, maintaining a moist wound environment, encouraging granulation and epithelial tissue formation, evaluating the need for debridement, and controlling infection. PMID- 17410827 TI - Current concepts in the management of delirum. AB - Delirium is a serious complication of physical illness that is commonly seen in the elderly individuals admitted to the hospital. It is associated with increased mortality, increased length of stay in the hospital, institutionalization and other complications. Multiple predisposing factors in the elderly them to make vulnerable delirium. The developing diagnosis of delirium is often missed and the condition is often poorly managed. There is increasing evidence suggesting that primary prevention of delirium is the most effective treatment strategy. This article presents current management for prevention and management of delirium. PMID- 17410828 TI - Falls--where do we stand? AB - Falls occur commonly in older persons and are the seventh leading cause of death. Falls are associated with functional deterioration and "fear of falling". Falls can be due to extrinsic factors such as poor lighting, throw rugs and other environmental hazards. Intrinsic causes of falls include physiological changes associated with aging, orthostatic hypotension, many medications, delirium, anemia, diabetes mellitus, Parkinson's disease, depression, cognitive impairment, syncope, partial complex seizures and vitamin D deficiency. Management of falls requires a multidisciplinary approach with a home assessment and modification where appropriate, a careful geriatric assessment, exercise programs focusing on balance, resistance and endurance exercise and adequate vitamin D replacement. All fallers should be assessed and treated for osteoporosis. The complexities of the causes and management of falls, make persons with frequent falls an ideal person to be referred for a geriatric consult. PMID- 17410829 TI - Andropause. AB - Andropause is a common part of aging in older men that affects a multitude of health parameters. It is imperative that male hypogonadism be evaluated and treated with hormonal replacement to avoid the development of frailty and effects on sexual health. This article will review the diagnosis and management of andropause. PMID- 17410830 TI - Evaluation of the elderly driver. AB - Driving is a daily activity for people across the country. Most Americans, including the elderly, consider it a right. Driving, however, is dangerous and can become more challenging as people age. While evaluating the elderly driver is not a simple task, it is a responsibility that must be shared by Missouri physicians. This paper describes the epidemiology of this problem, common mistakes made by elderly drivers, risk factors that can cause unsafe driving, and strategies physicians can use to evaluate their elderly patients' ability to drive. PMID- 17410831 TI - Evaluation of urinary incontinence and voiding dysfunction in women. AB - Urinary incontinence currently affects 33 million American women. In the next 30 years the population over 60 years will increase by 80-90%. This will result in a tremendous increase in patients suffering this condition. It is therefore not feasible for evaluation and treatment to be limited to specialists. Primary care physicians must be comfortable performing basic evaluations which will enable them to formulate a diagnosis and conservative treatment plan. This article outlines a practical approach. PMID- 17410832 TI - Epidemiology of methamphetamine abuse in Missouri. AB - Methamphetamine use has spread over the past decade from the West to other regions of the nation. Since 2000, Missouri has ranked first in clandestine laboratory incidents. The continuing threat of Mexican-produced methamphetamine tempers recent reduction of clandestine laboratory incidents in Missouri. There are a number of consequences related to the use of the drug and Missouri's healthcare professionals could potentially play key roles in prevention and treatment of the problem. PMID- 17410833 TI - Restoration challenges for urban rivers. AB - Urban sources account for significant quantities of important diffuse pollutants, and urban watercourses are typically badly polluted. As well as toxic metals, hydrocarbons including PAHs, and suspended matter, priority urban pollutants include faecal pathogens and nutrients. Can urban watercourses be restored by sufficient reductions in pollution loads? Case studies in the UK and Sweden provide insights and some grounds for optimism. A major trans-Atlantic review of the performance of best management practices (BMPs) is informing BMP planning. New approaches such as the maximisation of self-purification capacity in the receiving waters may also need to be developed, alongside BMPs at source. Other initiatives in Europe, USA and China, including collaborative projects, are trying to address the intractable issues such as persistent pollutants from transport and urban infrastructure. The challenge is daunting, but there are clear ways forward and future research needs are evident. PMID- 17410834 TI - Diffuse pollution associated with the mining sectors in South Africa: a first order assessment. AB - Internationally it has become recognised that diffuse source water pollution from mining activities severely affects the degradation of water quality especially with regards to acidification and metal loading. South Africa is facing major problems with regard to the management and treatment of contaminated mine water. Very little has been published for South Africa about the quantities and qualities of diffuse source water pollution by the mining industry. Furthermore the available information has not yet been compiled into a consolidated overview that presents the total picture. Some of the problems that limit the use of the available information and would necessitate further processing to normalise the data, derive from the fact that the investigations producing the information were done at different times, to different levels of detail and using different approaches. A further complicating factor is that data for some mining commodities may not be available and may necessitate further investigation. The overview of the quantities and qualities of non-point source effluent production by different sectors needs to be interpreted in terms of the effect the effluent can be expected to have on receiving water quality (both surface and groundwater). It would thus be necessary to categorise waste types according to their effect on water quality and synthesise the data to obtain an estimate of the threat that different sectors and sub-sectors pose to receiving water quality. PMID- 17410835 TI - Diffuse pollution from intensive agriculture: sustainability, challenges, and opportunities. AB - Global expansion and intensification of industrialized agriculture during the last 50 years was facilitated by the replacement of labor by imported chemicals and energy, thus changing the economics and the social fabric of rural communities as well as impairing water, air, and soil resources essential to sustaining food and fiber production in a world with an increasing appetite. To effectively understand and solve complex problems resulting from this agricultural revolution, expanded communications are needed at a variety of levels. It is critical for the technical community to communicate through greater interdisciplinary research among agronomists, soil scientists, hydrologists, ecologists, and others to reduce diffuse pollution from agriculture. Also, more effective translations of technical problems and solutions are needed to influence policy. Accurate advice is needed in spite of the uncertainties that scientists too often use to obscure useful information. Education will be needed for producers and conservationists to gain confidence that promised environmental responses will occur if solutions are to be implemented at more than experimental or demonstration scales. The search for comprehensive solutions to environmental degradation will require understanding the ultimate causes of pollution, not just the proximal causes. The ultimate causes will only be found by examining the systems that facilitate the release of contaminants to the environment such as the wholesale landscape changes that replaced grazing land with annual crops leading to increased leaching and runoff. Research and demonstration projects increasingly need collaborations among agronomists, livestock scientists, soil scientists, hydrologists, economists, sociologists and others who have a stake in the study of diffuse pollution and the outcomes of any proposed solutions. Partnerships developed at the working level where basic principles can be shared will help avoid the pursuit of impractical solutions when viewed from different perspectives. PMID- 17410836 TI - Grassed buffer strips as nitrate diffuse pollution remediation tools: management impact on the denitrification enzyme activity. AB - The management of grassed buffer strips proved to be an efficient remediation technique in controlling nitrogen losses to surface water. In south Belgium, agri environmental policies have encouraged farmers to seed buffer strips along rivers, in zones where the soil was previously devoted to agricultural production. We wanted to assess how important denitrification is in a buffer strip in comparison with a cropped field. The study investigated the denitrifiying enzyme activity (DEA) of two contiguous buffer strips with different management stories. The eastern part of the buffer strip was seeded in 1999. The western part of the buffer strip is a piece of crop field abandoned by the farmer 20 years ago and not managed for the last 10 years. This experimental study demonstrates that the denitrification enzyme activity in a riparian buffer strip is significantly higher than in the adjacent cropped field (3.67 and 2.12 mgNkg(-1)d(-1) respectively). The DEA was significantly different between the two buffer strips under comparison, assessing that the management of the buffer strips has a dominant effect on DEA. The old unmown buffer strip is potentially more efficient in the nitrate removal process than the 6-year-old seeded buffer strip. PMID- 17410837 TI - Development of a resource protection and waste strategy for water use by the agricultural sector. AB - The South African Department of Water Affairs and Forestry (DWAF) has started developing a strategy to regulate activities and water uses by the agricultural sector that could impact on the water resource quality. The aim would not be to over-regulate the sector, but to protect the water resource where necessary. Most of these activities constitute diffuse sources of potential pollution. The strategic process will start with investigative discussions with major stakeholders and determining the strategic context and current situation. The latter will consist of a detailed literature and stakeholder survey, and an evaluation of existing agricultural activities. The next steps of determining a vision and the setting of strategic objectives will be done with active participation by the major players. An action plan will be developed to achieve the set objectives. Important components of the strategy will be to: classify activities according to their risk to the water resource, taking into account the sensitivity of the water resource; set regulatory measures in accordance with the risk posed by the activity (measures could include the promulgation of regulations, general authorisations and/or issuing of licenses); harmonise and link the process with existing relevant processes and guidelines within DWAF and other government departments; review existing guidelines; sign agreements with relevant government departments and the agricultural sector; and provide training, built capacity and raise awareness during and after the process. PMID- 17410838 TI - Pesticide retention in an experimental wetland treating non-point source pollution from agriculture runoff. AB - Pesticide losses to the environment are unwanted due to possible environmental and health hazards. An experimental wetland is established to study the efficiency with respect to retention of sediments, nutrients and pesticides. Pesticides were applied on an arable soil plot in the watershed. Statistical analyses were carried out on three selected pesticides: propachlor, metalaxyl and chlorfenvinfoss. All pesticides were found in the experimental wetland, with peak concenttrations shortly after spraying. In 2003 pesticide retention varied from 11% to 42% and in 2004 retention varied from 19% to 56%. Comparing eight different wetland filters, we found that L6 and L8, with flagstones and straw, respectively, had a higher total pesticide retention than a standard Norwegian wetland (L4). When the compounds were treated separately, however, the picture was different. Statistical analyses showed that the treatments were signficantly different from zero in six of the wetlands for remowal of propachlor, for removal of metalaxyl none were significantly different, and for removal of chlorfenvinphos four treatments were significantly different. For the three compounds none of the relative treatments were significantly different from L4. Chemical properties of the pesticides could explain some of the behaviour in the watershed and in the wetland. PMID- 17410839 TI - Non-point sources and the waste discharge charge system. AB - The development of a waste discharge charge system (WDCS) in South Africa has been proposed to promote waste reduction and water conservation. The WDCS is based on the polluter pays principle and is designed such that the management of waste discharges achieves resource quality objectives (RQOs) at the minimum total cost to the catchment. Two charges are distinguished: first a charge for optimising use of the resource (incentive charge); and secondly, a charge for development and operation of mitigation measures in the resource (mitigation charge). The WDCS is applied to both point sources and non-point sources (NPS) of contamination. In the inclusion of NPS, the charge system distinguishes between registered and non-registered NPS, with the charge applied to the former group only, in the first instance. This paper introduces the WDCS by describing the principles and the theoretical basis of the WDCS, highlighting the link to RQOs as the benchmark of acceptable externalities. The paper explores the inclusion of NPS in the WDCS, describing: first, the principles of NPS inclusion in the WDCS; secondly, the types of NPS included in the present version of the WDCS; thirdly, the methodology for charge estimation; and finally, non-registered NPS and their potential inclusion in future editions of the WDCS. The paper concludes with remarks and challenges facing the first edition of the WDCS. PMID- 17410840 TI - Efficacy of constructed wetlands to mitigate non-point source pollution from irrigation tailwaters in the San Joaquin Valley, California, USA. AB - The efficacy of using constructed wetlands (CWs) to sequester organic carbon and nutrients from irrigation tailwaters was studied in the San Joaquin Valley, California. Two CWs were monitored during the 2004 irrigation season, a new CW (W 1) and 10-year-old CW (W-2). Input/output waters from CW were collected weekly and analyzed for a variety of water quality contaminants. Organic carbon, nutrient and sediment retention efficiencies were evaluated from input/output concentrations. Characteristics of sediment were examined spatially at W-2. Results indicate that W-2 was more efficient at contaminant removal. Average particulate organic carbon retention, was 70+/-13% (mean +/-standard deviation) in W-2 and 48+/-32% in W-1. Chlorophyll-a, a measure of algal biomass, was higher at W-1, especially in input waters. Initially, output concentration of chlorophyll-a increased 15-fold in W-2, however over time, as emergent vegetation established, chlorophyll-a decreased to 35% of input levels. Average total N removal efficiency was 45 +/-18% for W-2 compared to 22 +/-32% in W-1. Total P removal efficiency was 72+/-14% at W-2 compared to 18+/-26% at W-1. CWs were most effective at removing total suspended solids, 84 +/-15% and 97+/-2% for W-1 and W 2, respectively. Results demonstrate that CWs are effective at capturing POC, sediment and nutrients from irrigation tailwaters. PMID- 17410841 TI - The ecological value of constructed wetlands for treating urban runoff. AB - The Sweetwater Authority's urban runoff diversion system (URDS) comprises constructed wetlands on a hillside between the town of Spring Valley and the Sweetwater Reservoir, California, USA. The URDS were designed to divert dry weather and first-flush urban runoff flows from the Sweetwater reservoir. However, these constructed wetlands have developed into ecologically valuable habitat. This paper evaluates the following ecological questions related to the URDS: (1) the natural development of the species present and their growth pattern; (2) the biodiversity and pollutant stress on the plants and invertebrates; and (3) the question of habitat provided for endangered species. The URDS wetlands are comprised primarily of rush (Scirpus spp.) and cattails (Typha spp.). This vegetative cover ranged from 39-78% of the area of the individual wetland ponds. Current analyses of plant tissues and wetland sediment indicates the importance of sediment sorption for metals and plant uptake of nutrients. Analyses of URDS water following runoff events show the URDS wetlands do reduce the amount of nutrients and metals in the water column. Invertebrate surveys of the wetland ponds revealed lower habitat quality and environmental stress compared to unpolluted natural habitat. The value of the wetlands as wildlife habitat is constrained by low plant biodiversity and pollution stress from the runoff. Since the primary Sweetwater Authority goal is to maintain good water quality for drinking, any secondary utilization of URDS habitat by species (endangered or otherwise) is deemed an added benefit. PMID- 17410842 TI - Determination of first flush criteria using dynamic EMCs (event mean concentrations) on highway stormwater runoff. AB - Recently the Ministry of Environment in Korea has developed the total maximum daily load program in accordance with the target pollutant and its concentration goal on four major large rivers. Since the program is largely related to regional development, nonpoint source control is both important and topical. Of the various nonpoint sources, highways are stormwater intensive land uses since they are impervious and have high pollutant mass emissions from vehicular activity. The event mean concentration (EMC) is useful in estimating the loadings to receiving water bodies. However, the EMC does not provide information on the time varying changes in pollutant concentration or mass emissions, which are often important for best management practice development, or understanding shock loads. Therefore, in this study a new concept, the dynamic EMC determination method, will be introduced to clearly verify the relationship between EMC and the first flush effect. Three monitoring sites in Daejeon metropolitan city areas were equipped with an automatic rainfall gauge and a flow meter for accumulating the data such as rainfall and runoff flow. The dynamic EMC method was applied to more than 17 events, and the improved first flush criteria were determined on the ranges of storm duration and accumulated rainfall. PMID- 17410843 TI - Instruments to reduce the leaching of heavy metals from building materials in the Netherlands. AB - In the Netherlands the leaching of heavy metals from metal building and constructing materials results in serious contamination problems in the water system. The most common sources of these heavy metals in construction materials are copper waterworks and roofs, zinc roofs, gutters and rain pipes, zinced steel, stainless steel, and lead sealing material. In urban waters the surface water and sediment standards are often exceeded. Although building and construction materials are certainly not the only source of heavy metals, they are an important part of the problem. This article focuses on six instruments that are in use in the Netherlands to try to reduce impact on the surface waters. In addition to this, national as well as international, a reconsideration of the risks and surface water standards for several heavy metals is considered. A balanced use of instruments can be considered as the application of a best practice. PMID- 17410844 TI - Runoff and windblown vehicle spray from road surfaces, risks and measures for soil and water. AB - Soil and surface water along roads are exposed to pollution from motorways. The main pollutants are polycyclic aromatic hydrocarbons (PAH), mineral oil, heavy metals and salt. These pollutants originate from vehicles (fuel, wires, leakage), wear and degradation of road surfaces and road furniture (i.e. crash barriers) and the application of de-icing salts. Runoff, vehicle spray and dry deposition disperse these contaminants into the soft shoulder (verges) of the roads and surface water to a measurable distance of about 50 up to more then 150 m from the road. Despite many monitoring programs, little is known about the risks of this diffuse pollution for soil and water quality and the geochemical and physical factors which determine these risks. Also little is known about the effects of possible measures. Therefore, extensive research has been carried out at two local motorways. Specific measurements on runoff, vehicle spray and effects of measures have been carried out for one year (13 months). This resulted in several new insights. The pollutants appear to adsorb effectively to natural soils. In vulnerable areas groundwater can be protected by adjusting the policy to removing the contaminated upper topsoil of the verges. Discharges of runoff into local surface water are not recommended. PMID- 17410845 TI - The influence of diffuse pollution on groundwater content patterns for the groundwater bodies of Germany. AB - Commissioned by Germany's Working Group of the Federal States on Water Problems (LAWA) the authors developed a procedure to define natural groundwater conditions from groundwater monitoring data. The distribution pattern of a specific groundwater parameter observed by a number of groundwater monitoring stations within a petrographically comparable groundwater typology is reproduced by two statistical distribution functions, representing the "natural" and "influenced" component. The range of natural groundwater concentrations is characterized by confidence intervals of the distribution function of the natural component. The applicability of the approach was established for 17 hydrochemical different groundwater typologies occurring throughout Germany. Based on groundwater monitoring data from ca. 26,000 groundwater-monitoring stations, 40 different hydrochemical parameters were evaluated for each groundwater typology. For all investigated parameters the range of natural groundwater concentrations has been identified. According to the requirements of the EC Water Framework Directive (article 17) (WFD) this study is a basis for the German position to propose criteria for assessing a reference state for a "good groundwater chemical status". PMID- 17410846 TI - A knowledge gap analysis on multi-scale predictive ability for agriculturally derived sediments under South African conditions. AB - Agriculture has been implicated as a major source of sediments in South Africa. The aim of the knowledge gap analysis was to understand the production and delivery components of agriculturally derived sediments under South African conditions and to assess the predictive ability to address the fate of these sediments from field to catchment scales. An overview is given of important erosion processes and erosion modelling applied in South Africa at the field and catchment scale. A limitation of the sediment models is that gully erosion is not simulated; therefore, the models should be complemented with gully erosion predictions if gullies are an important sediment source. Field-scale models inadequately predict sediment production localised at hydrologically sensitive areas as a result of saturation excess flow and/or throughflow. The discussion on erosion modelling reveals that more complex models have had limited application in South Africa because they require large and detailed data sets, and may have parameters that are difficult to measure or to estimate. A modelling framework is discussed which allows linking of sediment models requiring readily available data, gully erosion models/maps and the use of other techniques to assess the fate of agriculturally derived sediments from field to catchment scale. PMID- 17410847 TI - Analysis and modelling of land-based nutrient pollution by watershed models coupled with GIS: a case study from Turkey. AB - Diffuse pollution is usually temporally and spatially uncertain, and thus hard to analyze. In many cases, discretizing a diffuse source of pollution into individual point sources can ease diffuse pollution modelling and analysis, and therefore reduce high uncertainty especially in the spatial distribution of pollution loads. This is however a difficult task, since quite a number of sub drainage areas, with complex structures and land-use properties, has to be delineated. Watershed models can be used to delineate the sub-drainage areas in a watershed with high accuracy and locate the related outlets which connect the sub drainage areas to the main waterbody in a watershed. In this study, such an approach has been used on a case study to model the diffuse nutrient loads carried to streams that reach to a medium-sized lake in Turkey. The annual nutrient loads, which were calculated by using mathematical models, were then converted to a load-map with the help of a geographical information system. PMID- 17410848 TI - Area-differentiated modelling of P-fluxes in heterogeneous macroscale river basins. AB - A model (MEPhos) for the calculation of the total phosphorus output from diffuse sources by drainage, wash-off, groundwater outflow, soil erosion and rainwater sewers as well as from point sources is presented. The model is based on a pathway- and area-differentiated emissions approach and calculates mean long-term P-inputs to surface waters. Phosphotopes are used for spatial discretization and modelling of diffuse P-inputs. Based on the modelling results "hot spots" for high P-loads can be localized and a management option for the input reduction to surface waters can be proposed which are adapted to site properties. The applicability of the model is demonstrated for two macroscale river basins in Germany (- 13,000 km(2) each) with contrasting natural conditions and land use patterns. PMID- 17410849 TI - Water fluxes and diffuse nitrate pollution at river basin scale: coupling of agro economic models and hydrological approaches. AB - An integrated model system has been developed to estimate the impact of nitrogen reduction measures on the nitrogen load in groundwater and in river catchment areas. The focus lies on an area-wide, regionally differentiated, consistent link up between the indicator "nitrogen balance surplus" and nitrogen charges into surface waters. As a starting point of the analysis actual nitrogen surpluses in the soil were quantified using the agro-economic RAUMIS-model, which considers the most important N-inputs to the soil and N-removals from the soil through crop harvest. The most important pathways for diffuse nitrogen inputs into river systems are modelled with the water balance model GROWA. Additionally, the time dependent nitrogen degradation along the nitrogen pathways in soil and groundwater are modelled using the WEKU-model. The two selected river basins in Germany cover a variety of landscape units with different hydrological, hydrogeological and socio-economic characteristics. The results indicate a wide range of annual nitrogen surpluses for the rural areas between than 10 kg N ha( 1) x a(-1) and 200 kg N ha(-1) x a(-1) or more, depending on the type and intensity of farming. The level of nitrogen inputs into the surface waters is reduced because of degradation processes during transport in soil and groundwater. Policy impact analyses for a nitrogen tax and a limitation of the livestock density stress the importance of regionally adjusted measures. PMID- 17410850 TI - Assessment of point and non-point pollution associated with the power generation sector in South Africa. AB - Access to water and water availability remains a key factor in ensuring the sustainability of development in Southern Africa. The need for guidelines to improve management of this valuable resource, and to regulate pollutant discharge, is therefore of national interest. A new and growing threat to our natural water resources is non-point source (NPS) pollution. The important distinction between point pollution and NPS pollution is that the latter is difficult to identify and the entry point of contamination to resources is diffuse and not limited to a single location. NPS pollution associated with power generation includes, but is not limited to, atmospheric deposition resulting from emissions (air and water), leachate from coal storage piles and runoff from impervious areas which are covered with dust fallout from coal and ash handling operations. Emissions of primary concern are sulfur, nitrogen and mercury. PMID- 17410851 TI - The assessment of diffuse pollution from acid-mine drainage using an updated and revised diatom assessment procedure as an added-value bio-monitoring tool. AB - Three data sources (physico-chemistry, bio-monitoring and eco-toxicology) are currently used in South Africa to establish environmental water quality conditions. Environmental water quality in turn is key information required for the "ecological reserve determination" of river reaches. Bio-monitoring in South Africa has been limited in recent times mostly to the use of the SASS procedure which relies on invertebrates only. This paper describes the re-introduction of a diatom-based water quality assessment as an added-value bio-monitoring tool. A specific example is discussed citing the response of diatom assemblages to diffuse pollution from acid mine drainage and how effective diatoms are as indicators of ecological integrity and river recovery measured downstream of the area of impact. The advantages of applying this bio-monitoring technique over other biological measures are presented in the context of technological advances in rapid image processing, species identification and software applications of diatom-based water quality indices. The valuable records of the diatom assemblages of the past, held in the South African Diatom Collection at the CSIR (KwaZulu-Natal), can now be accessed and interpreted as historical environmental water quality reference points for several rivers in South Africa. PMID- 17410852 TI - Long-term monitoring and proposed diffuse pollution control of a tropical reservoir. AB - This paper presents the results of 30 years of water quality monitoring in a tropical water supply reservoir (Vargem das Flores, Brazil). This water body is subjected to eutrophication problems caused by point sources (discharge of untreated sewage) and diffuse pollution (agricultural use in the drainage basin). Emphasis is given here on the estimation of nutrient loads and on the study of the N/P ratio in the water body. In spite of the prevalence of high N/P values, there is a clear trend in the dominance of cyanobacteria in the phytoplankton, which contradicts results from research in temperate aquatic environments. Some restoration measures for Vargem das Flores Reservoir are currently being implemented: construction of wastewater treatment plants, control of recreational activities, erosion control by hydroseeding and use of natural wetlands. Finally some management strategies in order to prevent algae input in the water abstraction system are discussed. Examples of these techniques are the installation of plastic barriers and the construction of an air curtain device. PMID- 17410853 TI - Water quality estimation in consideration of pollutant runoff and internal production in Lake Biwa, Japan. AB - Lake Biwa is the largest lake in Japan, and water quality in the lake is heterogeneous. Therefore, it is important for water quality management that spatial distribution of water quality in the lake should be clearly understood. The objectives of this study are to show a methodology and to develop a simulation system to calculate COD distribution in Lake Biwa taking internal COD production into consideration. This study also aims to examine transition of COD in the lake using the simulation system. In the simulation system, runoff loads of COD from the Lake Biwa basin are calculated by Macro Model for each tributary. The external COD concentration in 233 inshore meshes of the Lake Biwa water surface was calculated using the runoff loads. The internal COD was calculated using relationships among limiting nutrients, chlorophyll-a and COD. Then, the spatial distribution of water quality in Lake Biwa was calculated both for the external and internal COD by spline technique. Simulations using the system were implemented for 1986-1998, and a clear difference in characteristics between a drought year and a flood year was shown. In the result, it was shown that the simulation system developed here was available to calculate COD distribution in Lake Biwa, and that it had the possibility to explain the recent phenomenon of COD increase in the lake. PMID- 17410854 TI - The dynamic water-sediment system: results from an intensive pesticide monitoring campaign. AB - To gain insight into the dynamics of pesticides in the different compartments of a river system, an intensive monitoring campaign was set up. An extensive dataset is useful when planning to model the fate of pesticides in river systems. In this study we focus on the Nil, a small, hilly basin situated in the central part of Belgium. Two automatic samplers were placed in the river basin, i.e., one at the mouth and one upstream, taking composite samples of water and suspended solids. Undisturbed sediment samples were taken by means of a macro-core and immediately frozen with CO2-ice. The samples were sliced and analysed for pesticides in pore water and on sediment. The results of the study revealed the dynamics of the different compartments of a river system. The water compartment showed hourly variations in pesticide peak concentrations, regularly exceeding the standards. The amount of pesticides transported by suspended solids increased after a rainfall event. The concentration of pesticides in pore water, measured on a monthly basis, followed the trends of the water compartment remarkably well, but in a significantly lower concentration. These observations may be explained by combined diffusion, biodegradation and chemical reactions occurring in the water column and the sediment. Further insight into the importance of each of these processes can be gained by dynamic modelling, an approach that is currently being followed. PMID- 17410855 TI - The application of bioremediation: reduction of metal concentrations in river water and COD in distillery effluent. AB - The major aim of this study was to evaluate and develop artificial bioremediation systems to reduce or remove metal pollutants from contaminated river water and to decrease the chemical oxygen demand (COD) in distillery effluent. Metals were extracted using the nitric acid digestion method, and the concentrations determined using inductively coupled plasma-atomic emission spectrometry. A decrease in metal concentrations was observed for most of the metals analysed in the river water after being pumped through the bioreactor system for approximately two weeks, e.g. Al concentration decreased from 0.75mg.l(-1) to 0.18mg.l(-) and for Ni, from 0.19mg.l(-1) to 0 mg.l(-l). In addition, the COD counts decreased from 2255 mg.l(-1) to a final value of < 150 mg.l(-1) in the distillery effluent. It could thus be concluded that the bioreactor system decreased the COD and metal concentrations in the distillery effluent as well as the river water, respectively. A bioreactor is being evaluated on-site at a wine cellar to reduce COD and will be constructed at a site along the Plankenbrug River. PMID- 17410856 TI - [Integrated Health Programs (PREVENIMSS)]. PMID- 17410857 TI - [Strategy of provision and assessment of preventive health services]. AB - This paper describes the strategy of provision and assessment of preventive health services that the Mexican Institute of Social Security (IMSS) established in 2002 and that for social communication purposes, was called PREVENIMSS. The strategy is composed by five programs: Children's Health, Adolescents' Health, Women's Health, Men's Health and Seniors' Health. The objectives of these programs are the following: to provide a group of actions to promote and protect health, according to their age, sex and risk factors; to incorporate new preventive actions in order to face demographic and epidemiologic transition; to promote the participation of population in health care; to integrate provision of preventive services in family medicine clinics, and to broaden their coverage and impact. The modernization process of the statistical information system is also described in this paper. It included several aspects, such as the establishment of an automated record of preventive actions, which allowed the generation of relevant and timely information to measure the coverage and both national and local impacts. PMID- 17410858 TI - [ENCOPREVENIMSS 2003, 2004 and 2005. 1. Methodology and studied population]. AB - In order to assess the progress of preventive health programs by measuring the coverage and other health indicators, ENCOPREVENIMSS (Health National Surveys) 2003, 2004 and 2005 were carried out. These surveys were applied nationwide to population affiliated to the Mexican Institute of Social Security, and were representative at a state level. The sampling frame was stratified, with several stages, by clusters, and had a minimum sample of 328 people per program group in each delegation. The applied questionnaires were previously designed and evaluated through a pilot test. They included specific questions for each program component per age group. In 2004, two sections were added, based on validated instruments to evaluate frequency of food intake and physical activity. A total of 79,797 people were surveyed: 117,884 in 2004, and 122,380 in 2005. A third of the population was insured, while the rest was beneficiary; about half the population was between 20 and 59 years old, and the proportion of people older than 60 increased almost 2% from 2003 to 2005. More than 90% of those who claimed to have a paid job were factory workers or employees. More than 90% of the population were assigned to a family physician in all the assessed years. The information obtained from ENCOPREVENIMSS is a valuable tool to conduct institutional policies for public health. PMID- 17410859 TI - [ENCOPREVENIMSS 2003, 2004 and 2005. 2. Advances in Children and Adolescents' Health Programs]. AB - INTRODUCTION: the National Survey of Coverage of Integrated Health Programs (ENCOPREVENIMSS) are probabilistic population-based surveys with delegation representativeness of insured population of the Mexican Institute of Social Security, belonging to different age groups. OBJECTIVE: to evaluate the coverage of the Integrated Health Programs for Children and Adolescents during 2003-2005. MATERIAL AND METHODS: a descriptive analysis of ENCOPREVENIMSS 2003, 2004 and 2005 was carried out, with regard to the components of Children and Adolescents' Health Programs, in the 37 delegations (states) of IMSS. Through a home sample, information of sociodemographic characteristics, use of health services and coverage of the main components of PREVENIMSS, were collected. The concentration of information and the data analysis took place at the Coordination of Integrated Health Programs. RESULTS: from 2003 to 2005, the coverage of both groups increased, mainly in children. The advances of coverage were different for each activity. The ones with best results were the incorporation to protocols of preventive attention, and vaccination, as well as weight and height measurement. The actions with lowest achievement were the detection of visual problems and the nutritional evaluation. CONCLUSIONS: coverage analysis is useful to ascertain the degree of application of the different preventive activities and it is also a good basis for planning. PMID- 17410860 TI - [ENCOPREVENIMSS 2003, 2004 AND 2005. 3. Advances in Women, Men and Seniors' Health Programs]. AB - This paper presents the results of the National Survey of Coverage of Integrated Health Programs (ENCOPREVENIMSS) 2003, 2004 and 2005, with regard to the coverage of Women, Men and Seniors' Health Programs. ENCOPREVENIMSS are nationwide probabilistic population-based surveys with delegation (state) representativeness of insured population of the Mexican Institute of Social Security. The sampling frame was stratified, with several stages, and by clusters. The surveys gathered information of sociodemographic characteristics, use of health services and coverage of the main components of the Health Integrated Programs (PREVENIMSS), as well as other complementary variables. For the purpose of this article, the main variables studied were those related to health promotion, weight, height and waist measurement, as well as other related to early detection of certain diseases. The results show an increase in the coverage during the period 2003 2005 in the three groups; however, the lowest were found in the group of men from 20 to 59 years old and the highest, in the group of seniors older than 59, except for those related to the detection of cervical and breast cancer, which were higher in women from 20 to 59 years old. Coverage analysis is useful to ascertain the degree of imposition of the different preventive activities and it is also a good basis for planning. PMID- 17410861 TI - [ENCOPREVENIMSS 2003. Prevalence of low weight, overweight, general obesity and central obesity]. AB - OBJECTIVE: to estimate the prevalence of malnutrition in the population attending the Mexican Institute of Social Security. MATERIAL AND METHODS: we studied a 25% subset of the National Survey of Coverage of Integrated Health Programs performed in 2003 to estimate weight and height with standardized procedures; in adults over 20, waist circumference was also measured. We considered central obesity as waist circumferences larger than 88 cm for females and 102 cm for males. Based on weight and height, the prevalence of low weight, overweight and general obesity was estimated; based on waist circumference, central obesity was assessed. RESULTS: the prevalence of low weight in men and women was, respectively, of 5.3 and 6.4% in children; 6 and 8.2% in adolescents; almost none in adults, and of 11.7 and 3.6% in seniors. On the other hand, we observed that the prevalence of overweight and obesity increased progressively in both sexes since the first decade of life up to 87% in females between 50 and 59 years old and 82% in males between 40 and 49. The risk of central obesity was very high in women older than 20 in all decades of life (36.6 to 74.2%). In men, this risk had lower prevalence (12 to 31.7%). In the northern part of the country, women presented more central obesity, while in the southeast, men did. CONCLUSION: this report highlights the alarming prevalence of malnutrition in the Mexican Institute of Social Security population. This information is relevant for future strategies aiming to prevent these serious health problems. PMID- 17410862 TI - [ENCOPREVENIMSS 2004. 5. Intake of healthy or potentially unhealthy food]. AB - OBJECTIVE: to know the prevalence of healthy or potentially unhealthy food intake in the population insured by the Mexican Institute of Social Security. MATERIAL AND METHODS: food intake was obtained from frequency questionnaires classified as healthy (HF) or potentially unhealthy food (PUF), and according to its nature and frequency consumption. The prevalences according to age groups were obtained for <1 year old and every five years until 19; then, they were recorded by decades, sex, location and place of residence, with or without coast. RESULTS: in the first year of age, the consumption of HF is low, and the intake of PUF starts, with industrialized juices and soft drinks. Between the first and the ninth year of age, the consumption of HF increases; however, the intake of PUF increases too; there is not a significant intake of red meat, eggs and fish. Throughout adolescence, HF intake and diversity increases, although PUF consumption is higher too. In adults and senior citizens, the kind of HF most frequently consumed were cereals and leguminous plants, but they barely ate fruits and vegetables. The HF of greatest consumption were those of animal origin, and the riskiest ones were very greasy daily products with saturated fat, as well as soft drinks. We found that the population living in the Pacific coast consumes more sea food than the one living in the Gulf coast. We also found that, as people get older, the intake of PUF decreases. CONCLUSION: in the first years of life, the proper intake of meat, eggs and fish should be promoted, as well as the consumption of HF and the decrease of food with refined sugar, saturated fat or salt. PMID- 17410863 TI - [ENCOPREVENIMSS 2004. 6. Men and women's pattern of physical activity]. AB - OBJECTIVE: to determine the prevalence of physical activity in population from 20 to 59 years old covered by the Mexican Institute of Social Security. MATERIAL AND METHODS: the prevalence of physical activity was estimated, classifying it according to its intensity, frequency and duration. Recommended physical activity was considered when moderate activity was done at least five days a week, for 30 minutes, or when vigorous activity was done three days a week, for 20 minutes. Insufficient activity was that with lower frequency or duration than recommended, and inactivity, when no activity was done. RESULTS: the sample included 30,270 women and 25,237 men. The global prevalence of recommended physical activity was 17.7%; the one of insufficient physical activity, 65.5%, and the one of physical inactivity, 16.8%. The prevalence of physical inactivity in men and women was 13.8 and 19.4%, respectively. The most frequently reported activities in men and women with recommended or insufficient activity levels were walking (44.3%), running or jogging (34.2%) and soccer (10%). Women and men who watched television very often had a prevalence of physical inactivity of 30.9 and 20.5%, respectively. CONCLUSIONS: the prevalence of recommended physical activity is low and that of inactivity is high. Some measures of intervention are needed in order to promote physical activity in adults. PMID- 17410864 TI - [ENCOPREVENIMSS 2003. Reproductive health]. AB - OBJECTIVE: to analyze the current situation of some aspects of reproductive health in population covered by the Mexican Institute of Social Security (IMSS), based on the results of the National Survey of Coverage of Integrated Health Programs (ENCOPREVENIMSS) 2003. MATERIAL AND METHODS: we collected and analyzed data on fertility, contraceptive practice and prenatal and puerperal care. For this purpose, we made specific files as follows: 5823 adolescents from 15 to 19 years old, 16,275 men from 20 to 59 years old and 16 866 women from 20 to 49 years old, of the total sample of 79,797 questionnaires of ENCOPREVENIMSS 2003. RESULTS: the specific fertility rate in adolescent females was registered in 40 live births for every thousand women in this age group. About 70% of men and 41.7% of women said they used some kind of contraceptive method in their first intercourse. The prevalence of contraceptive use among female adolescents was 56% and among women from 20 to 49 years old, it was 76.5%, with the largest proportions of definitive or high-continuity contraceptive methods: 50.1% for tubal occlusion and 19.9% for intrauterine device. Global fertility rate for the whole population covered by IMSS was 2.56. Prenatal care coverage was 97.1% in adolescents and 98.2% in women from 20 to 49 years old. CONCLUSIONS: the data of ENCOPREVENIMSS 2003 confirm a high prevalence of contraceptive use in female population covered by IMSS, very similar to what was observed in other national surveys and in the population covered by IMSS. Besides, women attending IMSS show a reduction of fertility that gets them closer to demographic replacement rates. PMID- 17410865 TI - [Preventable diseases by vaccination. Coverage and impact]. AB - OBJECTIVE: to evaluate the coverage and impacts of the vaccination component in Integrated Health Programs. MATERIAL AND METHODS: a descriptive study of secondary data analysis was carried out. We analyzed data generated by the Universal Vaccination Program (PROVAC) since 1991, the data compiled in the 2000 National Health Survey (ENSA) and the 2005 National Survey of Coverage of Integrated Health Programs (ENCOPREVENIMSS), as well as the IMSS Annual Epidemiological Gazettes since 1973. RESULTS: the coverage of the basic schemes of the Extended Immunization Program (PAI) in children from one to four years old increased from 46.0% in 1990 to 92.5% in 1992 and 98.2% in 2005. The coverage with booster doses was substantially lower, ranging between 57.0% and 97.3% depending on the type of vaccine and the number of doses. The coverage also varied, although to a lesser extent, among public healthcare institutions. In relation to the impact of the Universal Vaccination Program and the PREVENIMSS strategy among IMSS affiliates, we distinguished at least three situations: (1) illnesses in which a drop in incidence predates the Universal Vaccination Program: tuberculous meningitis, diphtheria, whooping cough, and tetanus; (2) illnesses in which the drop in incidence is clearly related to the program: measles, rubella, mumps, and meningitis due to H. influenzae b. (3) illnesses for which the impact has not yet been assessed: hepatitis B. In addition to a drop in the incidence of immunopreventable diseases, we observed changes in the age distribution of cases. CONCLUSIONS: the PREVENIMSS strategy has not interfered with, and in some case has helped to consolidate the achievements of the Universal Vaccination Program. The results of this investigation point to the need to modernize the current system of evaluating coverage and expanding vaccination schemes with booster doses. PMID- 17410866 TI - [Mortality due to selected causes of infectious and chronic non-transmissible diseases, from 1991 to 2004]. AB - OBJECTIVE: to analyze the mortality behavior due to infectious diseases (acute diarrheal diseases, acute respiratory infections and lung tuberculosis) and chronic non-transmissible diseases (cervical cancer, breast cancer, diabetes mellitus and ischemic heart disease) in insured population of the Mexican Institute of Social Security (IMSS), in insured population user of the health services of IMSS and in uncovered population, from 1991 to 2004. MATERIAL AND METHODS: a descriptive study was carried out, in which the annual rates of national mortality for the selected causes were calculated by specific age groups in both insured and uninsured population. Death records were taken from the official registers of the National Institute of Statistics, Geography and Informatics and from the IMSS Mortality System. Population data were obtained from projections of the National Council of Population and IMSS records. RESULTS AND CONCLUSIONS: mortality due to acute diarrheal diseases, acute respiratory infections, lung tuberculosis and cervical cancer had a descending behavior, while breast cancer, diabetes mellitus and ischemic heart disease increased. Mortality for all causes analyzed was higher in uninsured population, except for diabetes mellitus, which was higher in insured population. This can be explained by the socioeconomic characteristics of the studied populations, as well as those of health services. PMID- 17410867 TI - [Maternal mortality at the Mexican Institute of Social Security, from 1991 to 2005. A period of changes]. AB - OBJECTIVE: to describe and analyze the levels, tendency, causes and preventability of maternal mortality occurred at hospitals of the Mexican Institute of Social Security (IMSS) in the period 1991-2005, in order to identify possible strategies and activities to get a faster reduction of this problem. MATERIAL AND METHODS: 3553 maternal deaths registered in medical units of IMSS between 1991 and 2005. This period was divided in two: from 1991 to 1997 and from 1998 to 2005, for comparison purposes. The analysis was made based on the conclusions reached by the Institutional Committees of Maternal Mortality Studies and the classification and codification according to the criteria of the 9th and 10th revisions of the International Disease Classification. The information on live births was obtained from the institutional information system named "Population and Services". RESULTS: in the fifteen-year studied period, the maternal mortality rate in hospitals decreased from 45.3 to 27 for every 100,000 live births (40.4%). The main causes of maternal death (preeclampsia-eclampsia, obstetrical hemorrhages, pulmonary tromboembolism) did not show any important changes in percentage values, even when the direct obstetric and specific rates for causes did show a reduction. By age groups, the highest maternal mortality rate was registered for women older than 35; the lowest one corresponded to adolescents. More than the third part of cases was considered as potentially preventable at admission in medical units. CONCLUSIONS: we identified that maternal mortality at IMSS hospitals tended to decrease in the analyzed period, particularly faster in the last 6 years. This was attributed to a better quality of medical care and timeliness in the use of medical services by population. PMID- 17410868 TI - [Analysis of the decrease in mortality due to cervical cancer at the Mexican Institute of Social Security, from 1991 to 2005]. AB - OBJECTIVE: to analyze the decrease in mortality due to cervical cancer in insured population of the Mexican Institute of Social Security (IMSS) and its relation to changes in the organization of the Program for Prevention and Control of Cervical Cancer (PCCaCu). MATERIAL AND METHODS: a descriptive study in which the annual rates of national mortality due to cervical cancer was analyzed from 1991 to 2005 was carried out. Information of deaths due to cervical cancer was obtained from the IMSS Mortality System (SISMOR), a database that identifies deaths occurred in insured population user of IMSS. The tendency was compared in three periods: I. 1991-1995, reorganization of the original program, II. 1996-2000, transition, and III. 2001-2005, incorporation to PREVENIMSS strategy. RESULTS: between 1991 and 2005, the mortality rate due to cervical cancer by 100 thousand women older than 24 years, decreased from 16.2 to 8.9 (45.1%). Between 1991 and 1995, the mortality rate increased 4.9%; between 1996 and 2000, it decreased 27% and in the last period, 2001-2005, it decreased 29.4%. The only differences that were statistically significant were the ones between the transition and the consolidation periods. The greatest decrease was registered when the PCCaCu was incorporated to PREVENIMSS strategy. CONCLUSIONS: the acceleration in the decrease of mortality rate due to cervical cancer can be explained by the reorganization and expansion of PCCaCu to target population and its incorporation to PREVENIMSS. PMID- 17410869 TI - [The role of sterols in morphogenetic processes and dimorphism in fungi]. AB - The review considers the fundamental biological problem of fungal dimorphism as an adaptive reaction to adverse impacts. Primary attention is paid to sterols, phospholipids, storage lipids, and fatty acids. The structural and biological functions of sterols are considered, as well as their role in membrane stabilization under stress and their relation to morphogenetic processes in mycelial fungi, of which many are pathogenic. Data on the biosynthesis of the main fungal sterol, ergosterol, are presented, as well as data on the inhibitors of this process and on the mutants deficient in its particular stages. Ergosterol biosynthesis is also considered in terms of its relation to the composition of the fungal cell wall, which is the cell shape-determining structure, and to the intensity of chitin synthesis, a process in which azole derivatives play a role. Data obtained by the authors are presented that show the role of changes in the composition of sterols, phospholipids, storage lipids, and unsaturated fatty acids of resting cells in the induction of yeastlike growth in mucoraceous fungi. PMID- 17410871 TI - [Synthesis citric and isocitric acids by Yarrowia lipolytica during yeasts growth on rapeseed oils]. AB - The native strain Yarrowia lipolytica VKM Y-2373 grown in a complete medium exhibited the maximum lipase activity at the concentration of rapeseed oil of at least 5.0 g/l. In the course of yeast growth, no considerable changes were observed in the glycerol concentration, the proportions of the major free fatty acids formed via oil hydrolysis, or the fatty acid composition of oil. Under nitrogen limitation of cell growth, the accumulation of citric acids reached 77.1 g/l with predominance of isocitric acid at pH 6.0, whereas at pH 4.5, almost equal amounts of citric and isocitric acids were produced. Cultivation of the mutant strain Y. lipolytica N 1 at pH 4.5 resulted in the predominant accumulation of citric acid (66.6 g/l) with an insignificant amount of isocitric acid. In the period of intense acid synthesis, high production of lipase was observed. PMID- 17410870 TI - [Dependence of the intracellular concentrations of univalent ions and hydrogenase activity on the salt composition and pH of the medium in the haloalkaliphilic sulfate-reducing bacterium Desulfonatronum thiodismutans]. AB - It has been shown that the intracellular concentrations of Na+, K+, and Cl- ions in Desulfonatronum thiodismutans depend on the extracellular concentration of Na' ions. An increase in the extracellular concentration of Na+ results in the accumulation of K+ ions in cells, which points to the possibility that these ions perform an osmoprotective function. When the concentration of the NaCI added to the medium was increased to 4%, the concentration gradient of Cl- ions changed insignificantly. It was found that D. thiodismutans contains two forms of hydrogenase--periplasmic and cytoplasmic. Both enzymes are capable of functioning in solutions with high ionic force; however they exhibit different sensitivities to Na+, K+, and Li+ salts and pH. The enzymes were found to be resistant to high concentrations of Na+ and K+ chlorides and Na+ bicarbonate. The cytoplasmic hydrogenase differed significantly from the periplasmic one in having much higher salt tolerance and lower pH optimum. The activity of these enzymes depended on the nature of both the cationic and anionic components of the salts. For instance, the inhibitory effect of NaCl was less pronounced than that of LiCl, whereas Na+ and Li+ sulfates inhibited the activity of both hydrogenase types to an equal degree. The highest activity of these enzymes was observed at low Na+ concentrations, close to those typical of cells growing at optimal salt concentrations. PMID- 17410872 TI - [Synthesis of the exopolysaccharide ethapolan on a mixture of energy-deficient growth substrates]. AB - Intensification of the synthesis of the microbial exopolysaccharide ethapolan by Acinetobacter sp. B-7005 was shown to occur on a mixture of energy-deficient growth substrates (acetate + glucose). When the bacterium grew on the substrate mixture, both substrates were utilized simultaneously; acetate was taken up by means of active transport at the expense of the energy of the proton-motive force. When acetate was present in the form of a sodium salt, the activities of acetyl-CoA synthetase and phosphoenolpyruvate synthetase (the key enzyme of gluconeogenesis) were tenfold higher than in the presence of potassium acetate, and the indexes of ethapolan synthesis were two times higher. The positive effect of Na+ on ethapolan synthesis is supposed to consist in the creation of ion gradients on the membrane, necessary for the generation of the proton-motive force. Simultaneous functioning of the glyoxylate cycle and pyruvate carboxylase reaction, as well as an increase in the activity of isocitrate lyase, malate synthase, and phosphoenolpyruvate synthetase, provide evidence of increased gluconeogenesis in the presence of the acetate + glucose mixture (as compared to gluconeogenesis on the corresponding monosubstrates). PMID- 17410873 TI - [Halo- and psychrotolerant Geomyces fungi from arctic cryopegs and marine deposits]. AB - Comparative characterization of Geomyces isolates was performed. The isolates were obtained from Arctic cryopegs and the surrounding ancient marine deposits, from nonsaline permafrost soils, and from temperate environments. Microbiological (cultural and morphological) and molecular criteria were used to confirm the identification of the isolates as Geomyces pannorum. The isolates from cryopegs and surrounding marine deposits were shown to differ from those obtained from nonsaline soils and temperate environments in their ability to grow at negative temperatures (-2 degrees C) under increased salt concentration (10%). The results are discussed in relation to the possible inheritance of the adaptive characteristics acquired in specific environments. PMID- 17410874 TI - [Anionic carbohydrate-containing cell wall polymers of Streptomyces melanosporofaciens and related species]. AB - The structures of cell wall anionic carbohydrate-containing polymers in Streptomyces melanosporofaciens VKM Ac-1864T and phylogenetically close organisms S. hygroscopicus subsp. hygroscopicus VKM Ac-831T, S. violaceusniger VKM Ac-583T, S. endus VKM Ac-1331, S. endus VKM Ac-129, and S. rutgersensis subsp. castelarensis VKM Ac-832T--have been comparatively studied by chemical and NMR spectroscopic methods. The natural polymer of a new, previously unknown structure, Kdn (3-deoxy-D-glycero-Dgalacto-non-2-ulopyranosonic acid) with beta galactose residues at C-9, has been found in the cell walls of all the strains under study. The cell walls of all the studied organisms contain three teichoic acids (TA): a predominant TA (1,3-poly(glycerol phosphate) with N-acetylated alpha-glucosaminyl substitutes by C-2 of glycerol, and minor TAs, 1,3- and 2,3 poly(glycerol phosphate) polymers without substitution. Their chains have O acetyl and O-lysyl groups. Microorganisms of the above-mentioned species differ in the number of alpha-glucosaminyl substitutes and in the degree of their acetylation in the predominant teichoic acid. PMID- 17410875 TI - [Reaction of microorganisms to the digestive fluid of the earthworms]. AB - The reaction of soil bacteria and fungi to the digestive fluid of the earthworm Aporrectodea caliginosa was studied. The fluid was obtained by centrifugation of the native enzymes of the digestive tract. The inhibition of growth of certain bacteria, spores, and fungal hyphae under the effect of extracts from the anterior and middle sections of the digestive tract of A. caliginosa was discovered for the first time. In bacteria, microcolony formation was inhibited as early as 20-30 s after the application of the gut extracts, which may indicate the nonenzymatic nature of the effect. The digestive fluid exhibited the same microbicidal activity whether the earthworms were feeding on soil or sterile sand. This indicates that the microbicidal agents are formed within the earthworm's body, rather than by soil microorganisms. The effect of the digestive fluid from the anterior and middle divisions is selective in relation to different microorganisms. Of 42 strains of soil bacteria, seven were susceptible to the microbicidal action of the fluid (Alcaligenes.faecalis 345-1, Microbacterium sp. 423-1, Arthrobacter sp. 430-1, Bacillus megaterium 401-1, B. megaterium 413-1, Kluyvera ascorbata 301-1, Pseudomonas reactans 387-2). The remaining bacteria did not die in the digestive fluid. Of 13 micromycetes, the digestive fluid inhibited spore germination in Aspergillus terreus and Paecilomyces lilacinus and the growth of hyphae in Trichoderma harzianum and Penicillium decumbens. The digestive fluid stimulated spore germination in Alternaria alternata and the growth of hyphae in Penicillium chrysogenum. The reaction of the remaining micromycetes was neutral. The gut fluid from the posterior division of the abdominal tract did not possess microbicidal activity. No relation was found between the reaction of microorganisms to the effects of the digestive fluid and the taxonomic position of the microorganisms. The effects revealed are similar to those shown earlier for millipedes and wood lice in the following parameters: quick action of the digestive fluid on microorganisms, and the selectivity of the action on microorganisms revealed at the strain level. The selective effect of the digestive gut fluid of the earthworms on soil microorganisms is important for animal feeding, maintaining the homeostasis of the gut microbial community, and the formation of microbial communities in soils. PMID- 17410876 TI - [The pleiotropic nature of rib80, hit1, and red6 mutations affecting riboflavin biosynthesis in the yeast Pichia guilliermondii]. AB - The yeast Pichia guilliermondii is capable of riboflavin overproduction under iron deficiency. The rib80, hit1, and red6 mutants of this species, which exhibit impaired riboflavin regulation, are also distinguished by increased iron concentrations in the cells and mitochondria, morphological changes in the mitochondria, as well as decreased growth rates (except for red6) and respiratory activity. With sufficient iron supply, the rib80 and red6 mutations cause a 1.5 1.8-fold decrease in the activity of such Fe-S cluster proteins as aconitase and flavocytochrome b2, whereas the hitl mutation causes a six-fold decrease. Under iron deficiency, the activity of these enzymes was equally low in all of the studied strains. PMID- 17410877 TI - [Obtaining and characterization of DNA-containing micromummies of yeasts and gram positive bacteria with enhanced cell wall permeability: application in PCR]. AB - The procedure of obtaining DNA-containing cell envelopes ("micromummies") of bacteria, yeasts, and fungi using chaotropic salts has been developed previously and the possibility of their direct application in PCR has been demonstrated. The fine structure of micromummies has been studied by electron microscopic methods. This work has demonstrated that additional treatment of micromummies of yeasts and gram-positive bacteria with proteinase K results in hydrolytic degradation of cell proteins and drastic enhancement of cell wall permeability for macromolecules (DNA). Thus, the efficiency of PCR ex situ using resultant micromummies after washing off the products of protein hydrolysis and proteinase K can be increased. The results of electron microscopic study of ultrathin sections of yeasts (Pichia pastoris, Saccharomyces cerevisiae) and gram-positive bacteria (Micrococcus luteus, Arthrobacter globiformis, Bacillus subtilis) support the biochemical data that treatment with chaotropic salts and proteinase K results in the loosening of microbial cell walls and in a decrease in the intracellular protein content. At the same time, cell walls generally maintain their integrity (continuity) and initial spherical or rodlike shape. The optimal modes of treatment of the cells of different microbial species with chaotropic salts and proteinase K have been selected to obtain permeabilized cell envelopes containing denatured or native DNA. PMID- 17410878 TI - [Hybrid formation and copulation activity in the yeast Pachysolen tannophilus]. AB - The copulation activity and hybrid formation efficiency have been studied in the xylose-assimilating yeast Pachysolen tannophilus. It was shown that the presence of 2% D-glucose, 0.5% yeast extract, and 2% agarose in the growth medium provided for the highest frequencies of hybrid formation. Atypical hybrid cultures similar in morphophysiological characteristics to native haploid strains of P. tannophilus were revealed in the course of hybridization. The genesis mechanism of such cultures and the reasons for the restricted applicability of hybridological analysis to genetic studies of P. tannophilus are discussed. PMID- 17410879 TI - [Vitamin B12-independent strains of Methylophaga marina isolated from Red Sea algae]. AB - Two strains (KM3 and KM5) of halophilic methylobacteria isolated from Red Sea algae do not require vitamin B12 for growth and can use methanol, methylamine, dimethylamine, trimethylamine, dimethyl sulfide, and fructose as sources of carbon and energy. The cells of these strains are gram-negative motile monotrichous (strain KM3) or peritrichous (strain KM5) rods. The strains are strictly aerobic and require Na+ ions but not growth factors for growth. They are oxidase- and catalase-positive and reduce nitrates to nitrites. Both strains can grow in a temperature range of 4 to 37 degrees C (with optimal growth at 29-34 degrees C), at pH between 5.5 and 8.5 (with optimal growth at pH 7.5-8.0), and in a range of salt concentrations between 0.5 and 15% NaCl (with optimal growth at 5 9% NaCl). The phospholipids of these strains are dominated by phosphatidylethanolamine and phosphatidylglycerol and also include phosphatidylcholine, phosphatidylserine, and cardiolipin. The dominant fatty acids are C(16:1omega7c) and C(16:0). The major ubiquinone is Q8. The cells accumulate ectoin, glutamate, and sucrose as intracellular osmoprotectants. The strains implement the 2-keto-3-deoxy-6-phosphogluconate-dependent variant of the ribulose monophosphate pathway. The G+C content of the DNA is 44.4-44.7 mol %. Analysis of the 16S rRNA genes showed that both strains belong to Gammaproteobacteria and have a high degree of homology (99.4%) to Methylophaga marina ATCC 35842T . Based on the data of polyphasic taxonomy, strains KM3 and KM5 are identified as new strains M. marina KM3 (VKM B-2386) and M. marina KM5 (VKM B-2387). The ability of these strains to produce auxins (indole-3-acetic acid) suggests their metabolic association with marine algae. PMID- 17410880 TI - [The new bacteriochlorophyll a-containing bacterium Roseinatronobacter monicus sp. nov. from the hypersaline soda Mono Lake (California, United States)]. AB - Two strains of pink-colored aerobic bacteriochlorophyll a-containing bacteria were isolated from aerobic (strain ROS 10) and anaerobic (strain ROS 35) zones of the water column of Mono Lake (California, United States). Cells of the bacteria were nonmotile oval gram-negative rods multiplying by binary fission by means of a constriction. No intracellular membranes were detected. Polyphosphates and poly 1-hydroxybutyric acid were the storage compounds. Pigments were represented by bacteriochlorophyll a and carotenoids of the spheroidene series. The strains were obligately aerobic, mesophilic (temperature optimum of 25-30 degrees C), alkaliphilic (pH optimum of 8.5-9.5), and halophilic (optimal NaCl concentration of 40-60 g/l). They were obligately heterotrophic and grew aerobically in the dark and in the light. Respiration was inhibited by light at wavelengths corresponding to the absorption of the cellular pigments. The substrate utilization spectra were strain-specific. In the course of organotrophic growth, the bacteria could oxidize thiosulfate to sulfate; sulfide and polysulfide could also be oxidized. The DNA G+C content was 59.4 mol % in strain ROS 10 and 59 mol % in strain ROS 35. In their phenotypic properties, the new strains were close but not identical to the alkaliphilic bacterium Roseinatronobacter thiooxidans. The distinctions in the nucleotide sequences of the 16S rRNA genes (2%) and low DNA-DNA hybridization level with Rna. thiooxidans (22-25%) allow the new strains to be assigned to a new species of the genus Roseinatronobacter, Roseinatronobacter monicus sp. nov. PMID- 17410881 TI - [Isolation and characterization of nitrogen-fixing bacteria of the genus Azospirillum from the soil of a Sphagnum peat bog]. AB - The presence of nitrogen-fixing bacteria of the genus Azospirillum in the soils of acidic raised Sphagnum bogs is revealed for the first time. Three Azospirillum strains, B2, B21, and B22, were isolated as a component of methane-oxidizing enrichment cultures, whereas attempts to isolate them directly from peat samples have failed. The results of comparative analysis of the nucleotide sequences of 16S rRNA genes, DNA-DNA hybridization, and the analysis of the sequences of the functional genes encoding nitrogenase and ribulose-1, 5-bisphosphate carboxylase reveal that all the newly obtained strains can be classified as Azospirillum lipoferum. Yet, unlike A. lipoferum. the isolates do not require biotin and utilize sucrose, inositol, and glycerol for growth. The cell morphology of strain B2 differs from that of the type strain and strains B21 and B22. The results obtained indicate the variability of morphological, physiological, and biochemical properties in closely related Azospirillum strains and suggest the existence of metabolic relationships between methanotrophic bacteria and the representatives of the genus Azospirillum under peat bog conditions. PMID- 17410882 TI - [Comparative molecular biological analysis of the microbial community of the holocene and pleistocene deposits of Posol'skaya Shoal, Lake Baikal]. AB - The bacterial diversity was studied in sediment layers of Posol'skaya Shoal station (Southern Baikal) belonging to different periods. A set of primers specific to individual bacterial groups was used to analyze the 16S rRNA gene fragments. The bacterial diversity in the Holocene deposits was found to be higher than in the Pleistocene ones. In the upper sediments, a positive PCR reaction with bacterial primers and with specific cyanobacterial and archaebacterial primers was detected. The following phylogenetic groups were revealed in the microbial community of the surface horizon: green nonsulfur bacteria, delta-proteobacteria, beta-proteobacteria (Nitrospirae), alpha proteobacteria, acidobacteria, crenarchaeota, euryarchaeota, and groups of uncultured bacteria. From the DNA of the Pleistocene deposits, the PCR product was obtained only with bacterial primers. The representatives of the genus Pseudomonas were most closely related to the sequences obtained (95-97% homology). PMID- 17410883 TI - [Cell hydrophobicity as a criterion of selection of bacterial producers of biosurfactants]. PMID- 17410884 TI - [Phylogeny and genetic diversity of the Astragalus cicer root nodule bacterial symbionts]. PMID- 17410885 TI - [A new thermotolerant aerobic methanotroph from a thermal spring in Buryatia]. PMID- 17410886 TI - Fatigue of tooth-colored restoratives in aqueous environment. AB - The purpose of this study was to investigate the interaction between mechanical and chemical fatigue in resin composites and dental ceramics, and the effects thereof on fatigue resistance of tooth-colored restoratives. To this end, the fatigue fracture resistance of restoratives under dry and aqueous conditions were analyzed by a dynamic fatigue crack propagation test using beam-shaped specimens with a precrack. Fatigue crack propagation characteristics were expressed by the correlation between fatigue crack growth rate (da/dN) and stress intensity factor range (deltaK). In addition, fatigue crack growth threshold (deltaKth) was calculated. Following the fatigue test, a fractographic examination was performed using scanning electron microscopy. Fatigue crack initiation was retarded in resin composites under aqueous condition, but dental ceramics were susceptible to slow crack growth after crack initiation. SEM images of the fatigue facture surfaces reflected inorganic and organic filler particles of different sizes in composites and the bonding at crystal-glass interface in ceramics. It was concluded that water exerted different effects on the fatigue resistance of composites and ceramics. PMID- 17410887 TI - Effect of polymerization accelerator on dentin bonding of one-step bonding agent. AB - The aim of this study was to enhance the bond strength of one-step bonding agents to dentin. In particular, the focus was on using Catabrush the applicator system of AQ Bond Plus. Catabrush was supplemented with N-phenylglycine and aromatic sulfinate as polymerization accelerators, as N-phenylglycine was reportedly beneficial in improving the bond strength to dentin. The results indicated that the bond strength to dentin was significantly augmented and the photo polymerization as well as the chemical polymerization were both improved even in the moistened dentin when 1.0 wt% N-phenylglycine was added to AQ Bond Plus agent, hence implying significantly higher bond strength to dentin. It was therefore concluded that N-phenylglycine is useful as a polymerization accelerator to be adopted in the applicator system for one-step bonding agents. PMID- 17410888 TI - One-year clinical evaluation of five single-step self-etch adhesive systems in non-carious cervical lesions. AB - This study evaluated the clinical performance of five single-step self-etch adhesive systems over a one-year follow-up period in 98 restorations. Independent evaluations were performed at baseline (one week), after three months, six months, and one year, by two trained examiners using the modified US Public Health Service rating criteria. Color match, marginal integrity, anatomical form, surface roughness, marginal or interfacial staining, postoperative sensitivity, and secondary caries were evaluated. Results were analyzed using the Kruskal Wallis test at a significance level of p<0.05. After one year, the restorations were re-evaluated and 100% retention rate was recorded. No caries were detected in association with any of the restorations. Marginal adaptation was the only measure that showed a slight deterioration over time, and there were no significant differences in this factor among the adhesive systems tested (p=0.464). PMID- 17410889 TI - Study of the variation between CIELAB delta E* and CIEDE2000 color-differences of resin composites. AB - This study sought to assess the significance of the corrections introduced in the new CIEDE2000 color difference formula with respect to deltaE*ab. The purpose of which was to provide sounder knowledge, and hence more informed decision-making, about applying this new formula to dental resin composites. With two different hybrid composites, color differences were calculated between unpolymerized and polymerized resin composites, between polymerized resin composites of different thicknesses (1 and 2 mm), and between polymerized resin composites cured with halogen and LED light curing units (LCUs). The two formulas differed significantly, with VAB(E) (equal size) values higher than the inter-observer variability (VAB (E)=11%) and VAB(O) (original size) values greater than 25% for each of the data sets analyzed. Results obtained in this study agreed with and thus supported the recent recommendation of the Commission Internationale de l'Eclairage (CIE), whereby the new CIEDE2000 formula should be used to evaluate color differences of resin composites. PMID- 17410890 TI - Water durability of resin bond to pure gold treated with various adhesion promoting thiirane monomers. AB - Adhesion promoting monomers for precious metals, 2,3-epithiopropyl methacrylate (EP1MA), 4,5-epithiopentyl methacrylate (EP3MA), 9,10-epithiodecyl methacrylate (EP8MA), 10,11-epithioundecyl methacrylate (EP9MA), 9,10-epithiodecyl 4 vinylbenzoate (EP8VB), 2,3-epithiocyclohexyl methacrylate (EPCHMA), and 3,4 epithiobutyl 2,2-bis(methacryloyloxymethyl)-propionate (EP2BMA), were used as surface treatment agents for pure gold to improve the durability of resin bonds against water. Treated specimens were butt-jointed together with MMA-PMMA resins, and tensile bond strength was measured after 2,000 thermal cycles in water. Bond strength to precious metal alloys was also determined under the same condition to clarify the influence of metal composition. The adhesion performance of thiirane monomers to pure gold highly depended on their chemical structure. EP3MA, EP8MA, EP9MA, EP8VB, and EP2BMA showed excellent performance, while EP1MA exhibited a moderate one and EPCHMA a poor one. The comparison of pure gold with precious metal alloys revealed the usefulness of pure gold as a standard, common adherend for bond strength evaluations. PMID- 17410891 TI - Effect of filler properties in composite resins on light transmittance characteristics and color. AB - The purpose of this investigation was to examine the effect of filler particle size and shape as well as filler content on light transmittance characteristics and color of experimental composite resins. A mixture of 30 mol% Bis-GMA and 70 mol% TEGDMA was prepared as a base monomer and to which a photoinitiator (camphorquinone) and a co-initiator (N,N-dimethylaminoethyl methacrylate) were added. Four different irregular- and spherical-shaped filler types with an average particle size of 1.9-11.1 microm were added to the mixture in three different filler contents of 20, 30, and 40 vol%. Light transmittance characteristics including light diffusion characteristics of the materials were evaluated. Color values and color differences among filler contents of the materials were also determined. Materials containing smaller and irregular-shaped fillers showed higher light transmittance and diffusion angle distribution with a sharper peak, as compared with those containing larger and spherical-shape fillers. It was also found that there was a significant correlation between the specific surface area of fillers and the color difference of the materials containing the fillers. Our results indicated that the shape of filler particles, as well as particle size and filler content, significantly affected the light transmittance characteristics--including light diffusion characteristics--and color of composite resins. PMID- 17410892 TI - Shear bond strength of a new fluoride-releasing orthodontic adhesive. AB - This study evaluated the shear bond strength of stainless steel brackets bonded to enamel with a new fluoride-releasing orthodontic adhesive system. A total of 140 extracted human bicuspids were randomly divided into four groups. Group I (Transbond XT) was a control group in which enamel was etched with phosphoric acid. For the remaining groups, enamel was conditioned with a self-etching primer (SEP): Group II (Transbond Plus), Group III (BeautyOrtho Bond), and Group IV (BeautyOrtho Bond + Salivatect). Stainless steel brackets were bonded to all tooth samples. After which, the samples were stored, thermocycled, tested, and statistically analyzed. Besides bond strength evaluation, the adhesive remnant index (ARI) was also evaluated. The shear bond strengths of Groups II, III, and IV were significantly lower than Group I, and Group II was significantly greater than that of Group III. Concerning ARI scores, no significant differences were found between the groups. Further, no enamel fracture was observed during shear bond test with the new SEP. In conclusion, when enamel was conditioned with the new SEP, the mean values of shear bond strength yielded were lower than when it was etched with 37% phosphoric acid. Nonetheless, these mean values were higher than the average suggested by Reynolds as optimum for clinical treatment. PMID- 17410893 TI - In vitro study of dentinal tubule occlusion with sol-gel DP-bioglass for treatment of dentin hypersensitivity. AB - DP-bioglass paste has been demonstrated to produce 60 microm of sealing depth on exposed dentinal tubules. However, the occlusive effect depended on a continuous placement of DP-bioglass paste on dentinal surface for three days. In a bid to fabricate highly reactive DP-bioglass particles, a sol-gel method was used together with HNO3, NaOH, and H3PO4 as catalysts. As a result, the application time of DP-bioglass paste was significantly reduced to 10 minutes. Percentage of tubular occlusion with DP-bioglass was 53.2-65.4%, while One Coat Bond and Seal & Protect yielded 51.3% and 41.2% respectively. Further, the average depth of tubular occlusion with DP-bioglass was 55.8-62.7 microm, while One Coat Bond and Seal & Protect produced 40.8 microm and 32.5 microm respectively. In conclusion, the best sealing performance of tubular occlusion was rendered by DP-bioglass catalyzed with HNO3. Its performance was significantly better than Seal & Protect, and was considered to exhibit the greatest potential in treating dentin hypersensitivity. PMID- 17410894 TI - Evaluation of the solubility of dental cements in artificial saliva of different pH values. AB - The aim of this study was to determine the solubility of dental cements in artificial saliva of different pH values. A total of 180 specimens--20 specimens (20 mm diameter x 1.5 mm thickness) for each dental cement-were prepared. After the specimens were weighed, they were set in the media with different pH values. After 24 hours, 72 hours, seven days, and 28 days, the specimens were taken out, dried in a desiccator, and re-weighed. Percentage of solubility at each time period was calculated as 100 per cent times weight loss divided by the initial weight of the specimen. Analysis of variance and LSD multiple comparison test were applied. It was found that cement type, storage time, and medium pH exerted statistically significant (p<0.001) effects on solubility. Among the tested cement types, zinc phosphate cement (Adhesor Carbofine) presented the highest solubility. In terms of medium pH, the cements seemed to be most stable in the medium with pH 7. PMID- 17410895 TI - Effect of acetic NaF solutions on fluoride-containing dental restorative materials. AB - The aim of this study was to evaluate the effect of acetic NaF solutions on fluoride-containing restorative materials. As the pH value of solution decreased, the degree of microhardness change in restorative materials increased- regardless of product. Dyract AP (DA) and F2000 (F2) (polyacid-modified resin composites) showed the greatest decrease in microhardness after immersion for three days. Similarly, as the pH value decreased, volumetric weight change (loss) increased in all products. DA and F2 showed the greatest--but similar-weight change in pH 3.5 solution among the products. In terms of color change, most specimens showed a slight color change after immersion for one and three days-regardless of pH value. However, F2 in pH 3.5 solution showed a noticeable color change (deltaE*=2.1). In terms of surface morphology, specimens in distilled water showed only minor surface modification. However, in pH 3.5 solution, DA and F2 showed randomly propagating cracks, while Solitaire 2 and Tetric Ceram (resin composites) lost many fillers less than 2 microm in size. PMID- 17410896 TI - Development of an easy-debonding orthodontic adhesive using thermal heating. AB - We produced experimentally a new bonding material that consisted of a mixture of a base resin (4-META/MMA-TBB resin adhesive) and thermoexpandable microcapsules for safe, easy debonding. Microcapsules in the base resin would start expansion at 80 degrees C, leading to a remarkable decrease in bond strength. Stainless steel brackets were bonded to bovine permanent mandibular incisors using bonding materials containing the microcapsules at different contents. After thermal cycling or heating, the shear bond strength of the brackets was measured. Shear bond strength of the bonding materials containing 30-40 wt% microcapsules decreased to about one-third or one-fifth that of the base resin on heating. Heating the brackets for eight seconds increased the temperature in the pulp chamber by 2 degrees C, which should not induce pulp damage. Results obtained suggested that the new bonding material should prove useful for removing brackets easily at the time of bracket debonding without any pain or enamel cracks, while maintaining the bonding strength during active orthodontic treatment. PMID- 17410897 TI - Cross-sectional TEM analysis of porcelain fused to gold-coated titanium. AB - This study investigated the interfacial microstructure between gold-coated titanium and low-fusing porcelain. The square surfaces of cast titanium split rods were sputter-coated with gold using a sputter coater at 40 mA for 1,000 seconds. Specimens were prepared for transmission electron microscopy (TEM) by cutting and polishing two pieces of the gold-coated split-rod specimens, which were glued and embedded in Cu tubes with an epoxy adhesive. TEM observation was also conducted for the gold-coated specimens after degassing and porcelain fusing. Due to the gold coating, intermetallic compounds of Au-Ti formed under the sputtered gold layer after degassing and porcelain fusing. Ti3Au and Ti3Al layers were also observed beneath the Au-Ti intermetallic compound layer. There was good adhesion of porcelain to the Au-Ti compound and Ti oxides without any gaps or formation of a Ti-deficient intermediate layer, which is normally observed at the titanium-porcelain interface. The results of this TEM study suggested that gold-sputter-coating the cast titanium surface produced a Ti-Au intermetallic compound and suppressed the formation of a Ti-deficient intermediate layer, resulting in improved adherence between porcelain and titanium. PMID- 17410898 TI - Effect of degradation of denture base resin on bond strength to relining resins. AB - The purpose of this study was to evaluate the effect of degradation of a denture base resin on bond strength to relining resins. Denture base resin specimens were immersed under various conditions to simulate degradation. Tensile bond strength of denture base resin after immersion to relining resins was measured with or without surface grinding. The bond strength of denture base resin without grinding after immersion to relining resin was significantly lower than that before immersion. However, bond strength was improved by surface grinding before bonding. These results suggest that bonding efficiency of denture base resin to relining resins was affected by immersion condition, which caused potential degradation of the denture base resin. PMID- 17410899 TI - Metallic pigmentation of human teeth and gingiva: morphological and immunological aspects. AB - The composition of metallic pigmentations in gingiva and dental roots was determined by means of transmission electron microscopy with energy dispersive x ray microanalysis. The systemic immune response to the metals found in the oral cavity was evaluated in 10 patients by using a modified lymphocyte proliferation test. Immunological results were compared with a group of five controls without metallic materials and pigmentation. Dense particles of various shapes and sizes, as well as of diverse extracellular and intracellular localization patterns, were detected in the pigmented lamina propria gingivae. Metallic deposits consisted predominantly of silver accompanied by selenium or sulfur or both. Besides, Ag, Au, Cr, Ni, Fe, Hg, Cu, and Ti were identified in dentinal tubules of teeth reconstructed with dental alloys. Nine patients with metallic pigmentations had a positive lymphocyte proliferative response to one or more metals present in their own metal reconstructions. Results of this study thus indicated that dental alloys-by virtue of their corrosion process-might pose a significant risk to immunologically susceptible patients. PMID- 17410900 TI - Multi-purpose bonding performance of newly synthesized phosphonic acid monomers. AB - Multi-purpose bonding performance of three kinds of newly synthesized phosphonic acid monomers was investigated. Methacryloxyalkyl or acryloxyalkyl phosphonoacetates of 6-MHPA, 6-AHPA, 10-MDPA were synthesized in 42.8-51.9% yields with a light yellow viscous liquid, and identified as new compounds by 1H NMR, IR, and elemental analysis. Conventional adhesive monomers, namely VBPA, 4 META, and 4-AETA, and CEBA-p-TSMo-t-BPMA initiator were also used. Seven experimental composite-type adhesive resins comprising these six kinds of adhesive monomers and None (control) with the initiator were prepared. Tensile bond strengths of adhesive resins to unetched ground enamel and dentin, ground porcelain, and sandblasted Ni-Cr alloy were measured at a crosshead speed of 1.0 mm/min. Results showed that except with VBPA, there were no significant differences among 6-MHPA, 6-AHPA, 10-MDPA, 4-AETA, and 4-META in bonding performance to the adherends (p<0.01). It was found that the new phosphonic acid monomers provided good multipurpose adhesion to all adherends tested. PMID- 17410901 TI - Effect of storage duration/solution on microshear bond strength of composite to enamel. AB - The aim of this study was to determine the effect of three storage solutions and two storage durations on microshear bond strength (microSBS) of a resin composite. Sixty non-carious human permanent molars were stored in three storage solutions (0.1% thymol, 10% formalin, and distilled water). Each tooth was separated mesio-distally into two parts. Specimens of the first part were stored for 24 hours, while specimens of the second part were stored for two months in the solutions. After each storage period, the enamel surface was covered with a composite resin in combination with an etch-rinse adhesive system. Specimens were then serially sectioned into sticks of 1 mm' bond area and subjected to microSBS test. There were no statistically significant differences between the two storage periods for each solution (p>0.05). The thymol solution group showed lower microSBS values than those of distilled water for both storage periods (p<0.05). As for the formalin group, its microSBS values were not statistically different from those of distilled water and thymol groups at each storage period (p>0.05). In conclusion, the thymol solution caused the microSBS of the resin composite to decrease when compared to both formalin and distilled water after 24 hours and two months. However, the microSBS of the resin composite was not affected by storage duration. PMID- 17410902 TI - Effect of curing method of a dual-cure resin cement on monkey pulpal reaction after bonding of tooth-colored inlay. AB - To compare the pulpal responses to light-cured and self-cured resin cements, cervical cavities were prepared in monkey's teeth, followed by application of etching gel and adhesive (Single Bond). A dual-cure resin cement (RelyX ARC) was applied, and hybrid composite inlays (Estenia) were bonded to the cavities. In one group, the cavities were photoirradiated for 20 seconds and the resin cement light-cured. In the other group, the resin cement was self-cured for six minutes without any photoirradiation. After experimental periods of seven, 28, and 70 days, histological features of pulp tissue were evaluated and compared. Results showed no significant differences in the histological features of the pulp tissues between the two curing methods. Both light-cured and self-cured resin cements showed acceptable biological compatibility with the monkey pulp. No bacterial penetration along the cavity walls was detected with either curing method. PMID- 17410903 TI - Development of metal-resin composite for dental magnet keepers. Part 1: effects of filler and 4-META contents on setting and flexural properties. AB - Nine kinds of experimental soft magnetic resin composites containing SUS447J1 stainless steel particles as filler were prepared. A UDMA/MAA resin with an MAA mole fraction of 0.67 was used as a matrix resin. The effects of three levels of 4-META content and three levels of filler content on the setting and flexural properties were studied. Working time and setting time significantly increased with increase of 4-META and filler contents. Flexural strength significantly increased with increase of 4-META content but with decrease of filler content. Elastic modulus simply increased with increase of both 4-META and filler contents. Although the flexural strength obtained was low, results were considered promising being the first step of this novel development. PMID- 17410904 TI - Enamel bonding of self-etch and phosphoric acid-etch orthodontic adhesive systems. AB - This study was conducted to examine the shear bond strengths of orthodontic brackets to ground enamel with two self-etching adhesives (Beauty Ortho Bond and Transbond XT) and two phosphoric acid-etching adhesives (Kurasper F and Orthomite Superbond). After the bonded specimens were stored in water at 37 degreees C for 24 hours, they were subjected to thermal cycling (TC) at 0 and 5000 cycles. Then, shear bond test was performed at a crosshead speed of 1 mm/min. Results indicated that the self-etching adhesives did not significantly differ in bond strength before and after TC (p>0.05), while the phosphoric acid-etching adhesives showed a significant reduction after TC (p<0.05). The predominant modes of bracket failure for the self-etching adhesives were at the enamel-resin interface, leaving less than 50% of the adhesive on the enamel surface. It was concluded that the self-etching adhesives exhibited stable bond strength to ground enamel without causing decalcification nor damage to the enamel surface. PMID- 17410905 TI - Comparison of the compressive strength of 3 different implant design systems. AB - The aims of this study were twofold: to compare the static compressive strength at the implant-abutment interface of 3 design systems and to describe the implant abutment connection failure mode. A stainless steel holding device was designed to align the implants at 30 degrees with respect to the y-axis. Sixty-nine specimens were used, 23 for each system. A computer-controlled universal testing machine (MTS 810) applied static compression loading by a unidirectional vertical piston until failure. Specimens were evaluated macroscopically for longitudinal displacement, abutment looseness, and screw and implant fracture. Data were analyzed by analysis of variance (ANOVA). The mean compressive strength for the Unipost system was 392.5 psi (SD +/-40.9), for the Spline system 342.8 psi (SD+/ 25.8), and for the Screw-Vent system 269.1 psi (SD+/-30.7). The Unipost implant abutment connection demonstrated a statistically significant superior mechanical stability (P < or = .009) compared with the Spline implant system. The Spline implant system showed a statistically significant higher compressive strength than the Screw-Vent implant system (P < or =.009). Regarding failure mode, the Unipost system consistently broke at the same site, while the other systems failed at different points of the connection. The Unipost system demonstrated excellent fracture resistance to compressive forces; this resistance may be attributed primarily to the diameter of the abutment screw and the 2.5 mm counter bore, representing the same and a unique piece of the implant. The Unipost implant system demonstrated a statistically significant superior compressive strength value compared with the Spline and Screw-Vent systems, at a 30 degrees angulation. PMID- 17410906 TI - Histologic evaluation of human bone integration on machined and sandblasted acid etched titanium surfaces in type IV bone. AB - The aim of this preliminary study was to evaluate the influence of a sandblasted acid-etched surface on bone-implant contact percentage (BIC%) as well as the bone density in the threads area (BD%) in type 4 bone after 2 months of unloaded healing. Five subjects (mean age = 42.6 years) received 2 microimplants each during conventional implant surgery in the posterior maxilla. The microimplants with commercially pure titanium surface (machined) and sandblasted acid-etched surface served as the control and test surfaces, respectively. After a healing period of 2 months, the microimplants and the surrounding tissue were removed and prepared for ground sectioning and histomorphometric analysis. One microimplant with a machined surface was found to be clinically unstable at the time of retrieval. Histometric evaluation indicated mean BIC% was 20.66+/-14.54% and 40.08+/- 9.89% for machined and sandblasted acid-etched surfaces, respectively (P=.03). The BD% was 26.33 +/-19.92% for machined surface and 54.84+/-22.77% for sandblasted acid-etched surface (P=.015). Within the limits of this study, the data suggest that the sandblasted acid-etched implant surface presented a higher percentage of bone-implant contact compared with machined surfaces, under unloaded conditions in posterior maxilla after a healing period of 2 months. PMID- 17410907 TI - Pilot study in the identification of stability values for determining immediate and early loading of implants. AB - This pilot study assessed the impact of implant stability criteria on implant loading time. Insertion torque (IT) and baseline implant stability quotients (ISQ) from resonance frequency analysis were recorded for 41 tapered-screw implants placed in the mandibles of 20 patients. Immediate (IL), early (EL) (6 weeks postoperative), or delayed (DL) (12 weeks postoperative) prosthetic loading was performed based on results and study criteria. Postoperative ISQ values were also recorded at 6 time intervals for the IL and EL groups and after healing for the DL group. All implants were definitively restored after 6 weeks of provisionalization. There were no failures, complications, or stability differences based on loading time. At week 12 of follow-up, ISQ values were slightly higher for IL implants compared with EL implants. IL and EL implants showed significantly higher cumulative ISQ values compared with DL implants. High IT and baseline ISQ values in all groups corresponded to high implant survival. Postoperative ISQ values in the IL and EL groups were less certain indicators of implant survival because of fluctuations relative to baseline values. Small sample size and inequitable patient distribution across groups skewed results. More research is needed before definitive results can be drawn. IL and EL were safely performed within the IT and ISQ ranges in this study, but it is unknown whether EL criteria would have also sufficed for IL. PMID- 17410908 TI - Using CAD-CAM technology for the full-mouth, fixed, retrievable implant restoration: a clinical report. AB - Computer-assisted design (CAD) and computer-assisted manufacture (CAM) are technologic advancements used in dentistry today. These technologies have been developed to fabricate a complete arch substructure from a solid piece of titanium for a fixed, screw-retained implant-supported prosthesis. This clinical report demonstrates a treatment approach which uses CAD-CAM technology to incorporate a milled, titanium framework with retrievable metal ceramic crowns. PMID- 17410909 TI - Bone healing at a failed implant site in a type II diabetic patient: clinical and histologic evaluations: a case report. AB - Placement of endosseous dental implants in diabetic patients may be compromised because of altered wound healing. Additionally there is no established timetable for implant healing in these patients. A case report is presented that evaluates implant healing at the site of a failed implant after its removal. This report documents bone remodeling in a diabetic patient 6 months after removing the failed implant. The prostheses was delivered within 4 months in the upper arch despite the complications during the healing period. PMID- 17410910 TI - The risks of repetition. PMID- 17410911 TI - Shoulders at work. PMID- 17410912 TI - New technology heralds hope. PMID- 17410914 TI - Not a lightweight decision. PMID- 17410913 TI - Pediatric strength training. PMID- 17410915 TI - Hands-on solutions. Traditional and high-tech therapies treat conditions of the hand. PMID- 17410916 TI - Surviving another round of medicare changes. PMID- 17410917 TI - Interview by Rogena Schuyler Silverman. PMID- 17410920 TI - Digging out from being buried alive: staff nurse leadership. PMID- 17410921 TI - New Web resource about pollutants and disease. PMID- 17410922 TI - Facing stigma: the challenge of AIDS. PMID- 17410923 TI - Identifying risk: the limitations of incident reporting. AB - Incident reporting is considered an important source of risk management data for identifying and addressing the causes of errors that occur in health-care organizations. However, the effectiveness of incident reporting in accurately identifying risk may be influenced by variables in the structure and design of incident reporting systems. Several variables, including data bias, system usability, non-standardized terminology and coding, and deficiencies in combined data analysis can reduce risk data accuracy. Until incident reporting systems are enhanced and an environment of safety is promoted, incident reports can provide only a glimmer of insight into organizational risk and quality improvement. Practice environments that support shared decision-making and collaborative care teams and provide stimulating learning opportunities can enhance professional nursing practice and inspire a culture of patient safety. PMID- 17410924 TI - Creating reporting and learning cultures in health-care organizations. AB - Patient safety has emerged as an important issue in Canadian health care, as reflected in the Canadian Council on Health Services Accreditation's patient/client safety goals. Achieving these goals calls for concerted efforts within health-care organizations. To assist nurse leaders in their efforts in developing a culture of safety that is receptive to reporting and learning from adverse events and near misses, the authors explore the challenges and provide four recommendations for action. By enacting these recommendations, nurse leaders can support the analysis and actions necessary to identify improvements that will create safer health-care environments. PMID- 17410925 TI - Donna Hagerty: a source of inspiration, information and hope for parents and their children. Interview by Tara Tosh-Kennedy. PMID- 17410926 TI - Unsafe student clinical performance calls for a systematic approach. AB - Patient safety is the responsibility of both the system and the individual practitioner. Unsafe incidents are a very real possibility when nursing students are preparing for their profession. The curriculum committee of the Nursing Education Program of Saskatchewan (NEPS) identified the need for a unified and consistent process related to students who demonstrate unsafe clinical performance. Many clinical teachers experience difficulty in identifying and making decisions related to students' unsafe performance. The authors describe the development of a systematic approach that was adopted by NEPS in June 2005 and is being used across all program years and sites. The approach provides students with a fair and just process and reflects the responsibility of the educational program to prepare graduates who will provide safe, competent care. PMID- 17410927 TI - The quality journey--advancing patient safety. PMID- 17410928 TI - [Dialectical behavior therapy for adolescents with anorexia and bulimia nervosa (DBT-AN/ BN)--a pilot study]. AB - Dialectical behavior therapy (DBT) was originally developed by Linehan (1993a, b) and modified by Miller et al. (1997) for suicidal adolescents with borderline personality features. Meanwhile, this therapy has also successfully applied in other adult clinical groups. The prior aim of the study is to evaluate the effectiveness of DBT for inpatient adolescents with anorexia and bulimia nervosa. In this pilot study (n=31) the efficacy of this treatment will be evaluated in a pre-post comparison. Different instruments will be used (SIAB, EDI-2, SCL-90-R, FBB). The first results are promising and we must hope that this new approach will improve the future treatment. PMID- 17410929 TI - [Legal probation of juvenile offenders after release from penal reformative training]. AB - Over recent years, there has been an increase in adolescent delinquency in Germany and Switzerland. In this context, the episodic character of the majority of adolescent delinquency is usually pointed out; however, numerous studies show high re-offending rates for released adolescents. The goal of this study is to examine the legal probation of juvenile delinquents after release from penal reformative training. In this study, the legal probation of adolescents committed to the AEA Uitikon, in the Canton of Zurich, between 1974 and 1986 was scrutinized by examining extracts from their criminal record as of 2003. The period of catamnesis was thus between 17 and 29 years. Overall, 71% of offenders reoffended, 29% with a violent or sexual offence. Bivariate logistic regression showed that the kind of offence committed had no influence on the probability of recidivism. If commitment to the AEA was due to a single offence (as opposed to serial offences), the risk of recidivism was reduced by 71% (OR=0.29). The results of the study show that young delinquents sentenced and committed to penal reformative training have a high recidivism risk. Furthermore, the results point out the importance of the evaluation of the offense-preventive efficacy of penal measures. PMID- 17410931 TI - [Regarding the child's right to information about his/her natural parents and regarding the parents' right to information about the actual maternity or paternity]. AB - It is true that the Federal Constitutional Court has explicitly affirmed the child's right to know his/her own descent; the parents' right to information regarding their natural paternity can be derived from the Constitution of the Federal Republic of Germany as well. This, however, does not determine who is entitled and at what point one may request information on the actual descent of a person. Neither does the right lay down from whom information may be requested or who could be sued for what kind of information. Further it leaves unanswered who would be in the position to enforce the judgement eventually. The adoption law is the only one containing legal provisions. The judicial decisions are unclear and inconsistent. It is still possible that there is a permanent difference between the legal and the actual paternity. In Germany, there is currently no room for determining genetic maternity that would differ from the legal maternity. Only older, grown-up children and men whose legal paternity has been eliminated in previous court proceedings are entitled to sue for information regarding their biological descent. Linked to this is the prohibition of secret paternity tests. In the question who is entitled to information regarding the actual descent (and whether there is such a right), financial, fiscal, health and purely moral interests of both, the parties involved and the public authorities must be balanced. In connection with a heterologous insemination, information regarding the genetic descent must not only be provided by the mother and the (legal) father but also by public authorities and other third parties, particularly physicians. Insufficient documentation on the part of the physician involves the risk regarding the liability for damages vis-a-vis the child who is entitled to receive information. PMID- 17410930 TI - [Parent satisfaction with a child and family-centered treatment program for primary headache in childhood and adolescence]. AB - Patient satisfaction represents a significant outcome criterion in the context of systemic psychotherapeutic therapy research. This study investigated parent satisfaction with a psychosocial treatment program (comprising three components: child group therapy, parents' evenings, systemic family sessions) for pediatric primary headache (diagnosed according to IHS criteria). 10 weeks after the end of the treatment program, the parents were sent a questionnaire containing open questions and ten-point numerical rating scales. The sample comprised n=48 families. The return rate was 89%. The qualitative content analysis showed a hierarchical category system consisting of 3 major categories, 7 main groups and 69 sub-categories. The parents stated that they were satisfied with (1.) the effects of the treatment program, (2.) the specific treatment techniques and the medical and psychosocial headache-related information provided, and (3.) the therapeutic relationship. The mean satisfaction for all three therapy components on the ten-point numerical rating scale was 8.1 with a standard deviation of 2.0 (child headache group: 8.5; parents' evenings: 8.2; family sessions: 7.5). The results are discussed with reference to methodological aspects: avoidance of ceiling effects and social desirability in measuring customer satisfaction, order effects of items, weighting of the significance of satisfaction levels as an outcome criterion. PMID- 17410932 TI - [Value(s)]. AB - After a short explanation of the word value, the (cultural) value of teeth, the economic evaluation of dentistry and the payment criteria are being presented. The specific situation of health care as a service which deviates in quite a few aspects from the standard demand-supply model is being pointed out. Attention is drawn to key characteristics of the liberal professions such as the obligation to perform to the best of one's ability, not to a specific result. Function classification appears to offer possibilities for cataloguing the wide variation in practice settings. The well-known wage calculation of Professor De Lembre is being reviewed. Subsequently the analysis of cost variation and induced demand by extra services and profile shaping is being elaborated. Cost-benefit-analysis is the concluding item. PMID- 17410933 TI - [Evidence-based medicine and its significance for primary health care]. AB - Evidence Based Medicine objectifies health care decision-making, but has its methodological restrictions and can be misused by economists, lawyers, politicians and other decision-makers. Guidelines are being formulated on how to apply EBM in primary care (general practice). PMID- 17410934 TI - [The phenomenon of the inducement of demand and various concepts in the economics of health]. AB - The article looks at the behaviour of practitioners and patients from an economic point of view, in particular at supplier-induced demand (SID). The objective is to be able to describe the factors which impact on the market for health care. To define the market for health care/dental care in general and the SID concept in particular, the following economic terminology is relevant. uncertainty, asymmetric information, moral hazard, and the agency relationship between practitioner and patient. Essentially SID results from the existence of asymmetric information (information "gap") on the health care market. The patient is less informed than the practitioner and cannot accurately assess his need for medical care. Hence decision-making authority is delegated to the practitioner who acts as the patient's agent to determine the optimal (amount of) care. This enables the practitioner to influence the demand for medical/dental care. If the practitioner is more interested in the rate of return than in the patient s need for care, a difference might arise between the care recommended by the practitioner and the care the patient would have asked for if he had the same level of information as the practitioner. This difference is called the amount of SID. PMID- 17410935 TI - [The inducement of demand in dental medicine in Belgium]. AB - Many factors are important for the existence of supplier-induced demand (SID). Not just the individual practitioner is responsible; his or her behaviour might be influenced by the specific organisation of the market for health care/dental care. The article investigates the characteristics of the Belgian market for dental care and analyses the way in which they might influence the occurrence of SID and/or overconsumption. Attention goes to the payment system of dentists in Belgium, the health insurance system, dentist density and the regulation of the sector. Also mentioned are the recall system and other (governmental) initiatives that may influence patients' and practitioners' behaviour. Some factors--not yet systematically investigated--may be relevant as well: personal characteristics of the dentist (gender in particular) and the way companies direct practitioners' and patients' behaviour. PMID- 17410936 TI - [Pyelonephritis treatment in children in 2007: current literature review]. AB - Four studies, including two being published as an abstract, have recently demonstrated the feasibility of oral treatment of pyelonephritis in children, with no increased risk of treatment failure, early urinary tract re-infection, or renal scars. To do so, the pediatrician must ensure that: (1) the patient does not appear toxic, has no vomiting; (2) there is no known severe obstructive or refluxing uropathy and (3) parents are deemed to be adherent to the treatment. If these criteria are fulfilled, the pediatrician can start an oral treatment with a 3rd generation cephalosporine for 10 to 14 days. Ambulatory follow-up is crucial, and persistance of fever after 3 days is a reason for a new outpatient visit, additional or supplementary imaging studies (renal ultrasonography) and eventually a switch to intravenous treatment. PMID- 17410937 TI - [Coeliac disease in children--an update]. AB - Coeliac disease (CD), considered as an autoimmune disease with genetic predisposition can occur at any age. Several types can be observed: a) typical cases presenting early in life with signs of intestinal malabsorption; b() atypical cases showing milder, often extraintestinal symptoms; c() silent cases that are occasionally discovered because of serological screening and (d) potential/latent cases showing isolated positivity of celiac serology at first testing and eventually the typical intestinal damage later in life. To date, the method of choice to make a diagnosis is the dosage of anti-transglutaminase IgA antibodies (including total IgA to prevent false negative results), followed by intestinal biopsy. Addition of moderate amounts of oats to a gluten-free diet does not prevent clinical or small bowel mucosal healing in children. PMID- 17410938 TI - [Utility of Gamma Interferon blood tests in the diagnosis of tuberculosis infection in children]. AB - Considering the ongoing difficulty with the interpretation of the tuberculin skin test and the importance of a correct interpretation of each situation in order to benefit from treatment, a lot of expectations are placed in the new Gamma Interferon blood tests. The aim of this article is to recall the basic principles of these tests and to review the evidence of their utility in paediatrics. In children, although the evidence is still insufficient, the blood tests can be useful in combination with the tuberculin skin test. PMID- 17410939 TI - [Of epigenetics and development]. AB - Epigenetics is defined as the study of the heritable changes in gene expression that are transmitted from a mother cell to a daughter cell or a parent to its progeny, that occur without modifications in DNA sequences. The epigenetic mechanisms of gene regulation include DNA methylation and post-translational modifications of histone tails such as acetylation. Generally speaking, the more methylated the DNA is, and the less acetylated the associated histones are, the less a gene will be expressed. Herein, we introduce the reader to the field of epigenetic reprogramming and its importance during embryonic development and gametogenesis. We also summarize the current litterature on environmental influences on epigenetic processes including the epigenetic transgenerational actions of endocrine disruptors and diet. PMID- 17410940 TI - [Epigenetic origin of diabetes and growth disorders]. AB - The understanding of genomic imprinting has made us realize that maternal and paternal contributions to the embryo are different. Disturbances during the imprinting process may lead to different pathologies due to an imbalance of gene expression either maternally or paternally derived. Known epigenetic diseases such as neonatal diabetes, growth retardation or overgrowth syndromes as well as cancer are better understood. It has become clear that environmental factors can be at the origin of such epigenetic changes. Careful analysis and diagnosis of epigenetic diseases are important for patient treatment and outcome. PMID- 17410941 TI - [Epigenetics and cancer]. AB - Since the early 80's, cancer research has been dominated by scientific breakthroughs demonstrating the genetic origin of cancer. Thousands of genetic alterations have been identified, affecting more than one hundred cell regulating genes. In the past ten years, our understanding of carcinogenesis has evolved: cancer is both a genetic and an epigenetic disease. Epigenetic modifications play a fundamental biological role in the initiation and progression of cancer by altering the expression of cell cycle regulation genes. Unlike genetic mutations, epigenetic modifications are potentially reversible. Thus, epigenetic inhibitors are currently evaluated as anticancer drugs. Moreover, DNA methylation study holds promise as biological marker for classification, diagnostic and prognostic purposes in clinical practice. PMID- 17410942 TI - [Surgical activity in a mass casualty situation: humanitarian relief mission in Pakistan]. AB - On October 8th 2005, a violent earthquake struck the northern region of Pakistan resulting in a large number of injuries and deaths. In response to this catastrophe, the Swiss Agency for Development and Cooperation (SDC) offered logistic support, including material and medical staff. After evaluating the needs, the SDC sent two surgeons to the city of Mansehra for a short term mission. At that site a college had been transformed into a field hospital by a multidisciplinary Pakistani team, already busy at work. Up to 200 casualties per day were brought to that center for a first surgical procedure consisting in debridement, application of traction, or external fixation. In total, more than 2000 people were treated during this two week mission. PMID- 17410943 TI - [Child labor in the world: a society problem]. PMID- 17410944 TI - [Who owns the H5NI strains? (1)]. PMID- 17410945 TI - [On France and neo-eugenism (3)]. PMID- 17410946 TI - [New neurons are born constantly in the human brain]. PMID- 17410947 TI - [Notes. Beasts or gods?]. PMID- 17410948 TI - Evaluation of the plaque removal efficacy of two commercially available dental floss devices. AB - OBJECTIVE: This single blind, crossover design, clinical trial provided a comparative assessment of the oral cleaning of two dental devices as demonstrated through the removal of dental plaque when used according to marketed product-use directions. METHODOLOGY: Twenty-six subjects, both male and female between the ages of 19 and 64 years, completed this two-cell crossover study. Subjects were assessed for whole mouth plaque levels, both before and after use of each of the randomly assigned floss devices, by assessing disclosed plaque using the Modified Turesky Plaque Index. The two floss devices were a Mint Floss Pick and a standard rolled floss. Subjects abstained from brushing the night before appearing in the clinic, and based upon meeting the minimum plaque criteria of 1.5 at the first phase of the crossover, used one of the two floss devices according to the directions for use found on the product packaging. Both pre- and post-device use plaque levels were recorded with calculations made of both the actual difference in plaque level, as well as the percent plaque removed. Data were subjected to an analysis of covariance (ANCOVA), and employed a model consistent with crossover design. RESULTS: Overall results from both phases of the crossover showed the Mint Floss Pick product removed 19.4% of the plaque. The standard rolled floss product removed 15% of the plaque. Both products removed statistically significant plaque when assessed versus pre-treatment levels. Although the ANCOVA identified a statistically significant difference between treatments favoring the Floss Pick product, further statistical examination revealed a significant sequence effect, which led to a conservative product comparison of equivalence. This analysis confirmed that the Floss Pick product was "at least as good as" the standard floss product for plaque removal. Additional analyses of various sites in the mouth, i.e., interproximal, anterior, posterior, etc., also confirmed similarity of performance for both products in this test. CONCLUSION: A two-way crossover assessment of the cleaning capability of a Mint Floss Pick product compared to a standard rolled floss product was performed through assessment of the removal of dental plaque. Results of this clinical investigation support the Floss Pick product to be "at least as good as" standard rolled floss in cleaning capability when both products were used according to their product-use directions. PMID- 17410949 TI - Extrinsic stain removal efficacy of a stannous fluoride dentifrice with sodium hexametaphosphate. AB - OBJECTIVE: Two randomized, six-week, double-blind, parallel group, clinical trials were independently conducted to compare the extrinsic stain removal efficacy of a novel whitening therapeutic dentifrice containing 0.454% stannous fluoride and sodium hexametaphosphate in a formulation with high cleaning silica, relative to a positive control whitening dentifrice. METHODOLOGY: Generally healthy adults, 56 for study 1 and 60 for study 2, with visible extrinsic tooth stain were enrolled in the studies. At the baseline visit, stain was assessed on the facial surfaces of the eight central and lateral incisors using the modified Lobene Stain Index. Oral soft and hard tissue examinations were also conducted. In each study, subjects were randomized to either the stannous fluoride + sodium hexametaphosphate toothpaste (Crest Pro-Health) or the positive control toothpaste (Colgate Total Plus Whitening) to use twice per day for six weeks. Stain and safety were reassessed at weeks three and six. RESULTS: Fifty-two and 58 subjects completed studies 1 and 2, respectively. In each study, there were no statistically significant differences in Lobene composite stain scores between the two treatment groups across all three visits. Both groups showed statistically significant reductions in Lobene composite stain scores at week three (p < 0.0001) and week six (p < 0.0001) relative to baseline. The percent of Lobene composite scores with a greater than 0.5 unit reduction from baseline at week six was 86% for study 1 and 97% for study 2 for the stannous fluoride + sodium hexametaphosphate dentifrice group. CONCLUSION: Collectively, these two stain removal clinical trials demonstrate the statistically significant extrinsic stain removal efficacy for the stannous fluoride + sodium hexametaphosphate dentifrice relative to baseline. There were no statistically significant differences between the stannous fluoride + sodium hexametaphosphate dentifrice and positive control treatment group. PMID- 17410950 TI - Silica as a reference for laboratory dentifrice assessment methods: multiple site collaborative study. AB - OBJECTIVE: This collaborative study was designed to collect data from multiple laboratories, using recognized abrasion and cleaning power methods, to assess the functionality, stability, and suitability of three Sident silica abrasives for consideration as references for ISO (International Standards Organization) abrasion and other dentifrice assessment methods. METHODOLOGY: This study included ISO and other physicochemical methods to characterize the abrasive powders, as well as ISO and related abrasion methods--Radioactive Dentin Method, Profilometry, and Copper Gravimetric, and two Cleaning Power methods--to test the functionality of the silica abrasives with the current ISO abrasion reference calcium pyrophosphate. RESULTS: The outcomes demonstrated that the collaborative study to measure the physicochemical characteristics of three Sident silica abrasive materials, stored at ISO and tropical conditions, found no change in the functionality as measured by these methods over the two-year test period. CONCLUSION: The three Sident silica abrasives demonstrated suitability to be considered as references for use with calcium pyrophosphate in the ISO 11609 Toothpaste Specification. PMID- 17410951 TI - A digital image analysis system for the assessment of tooth whiteness compared to visual shade matching. AB - OBJECTIVE: To assess tooth whiteness clinically using an image analysis system and a whiteness algorithm to compare with visual shade matching. METHODOLOGY: Images of the maxillary anterior teeth of 20 subjects were obtained using an image analysis system adapted for tooth whiteness measurement. Red, green, and blue values from each image, and Commision Internationale de l'Eclairage whiteness index (CIE WI) values were calculated. A visual shade match was also carried out using a Vitapan 3D Master 26-tab shade guide. RESULTS: The range of CIE WI values for image analysis and visual shade matching was 32.86-85.91 and 32.68-79.84, respectively. Limits of agreement between methods were +17.71 and 14.60 CIE WI units. The 95% confidence interval of the difference was calculated as -0.176 to +3.083. A two-tailed Student t-test with a 95% confidence level showed significant differences (p = 0.028) between the paired CIE WI values of each tooth for the two measurement techniques. As the accuracy and reliability of the image analysis system was found to be high in previous in vitro studies, the differences between the two methods were probably related to the inherent subjectivity of visual shade matching. CONCLUSION: The adapted digital image analysis system could be used as an alternative to, or in conjunction with visual shade matching. PMID- 17410952 TI - Effect of a stannous fluoride dentifrice on plaque formation and removal: a digital plaque imaging study. AB - OBJECTIVE: A repeated measures digital imaging technique (Digital Plaque Image Analysis-DPIA) was used to assess the antiplaque effectiveness of a 0.454% stannous fluoride dentifrice. METHODOLOGY: Subjects were qualified to enter the study based upon a previous assessment of dental plaque accumulation while brushing with a standard (non-antimicrobial) fluoridated dentifrice. Fourteen qualifying subjects were entered into a cross-over study design, including the use of a standard 0.243% sodium fluoride (NaF) dentifrice, or a 0.454% stannous fluoride (SnF,) dentifrice. Cross-over periods were separated by a one-week washout. During the assigned treatments, subjects visited the imaging laboratory on six separate days over two weeks, reporting each day for three separate assessments of plaque accumulation, as assessed by DPIA. On each "grading day," subjects entered for plaque assessments prior to morning tooth brushing, and without consuming any food or beverages. Following this morning "pre-brushing" assessment, subjects brushed with the assigned dentifrice and then had plaque immediately reassessed (morning post-brushing). Subjects also reported to the imaging clinic for an afternoon plaque regrowth assessment. Subjects were again required to avoid food and drink for one hour prior to this measurement. RESULTS: The SnF2 dentifrice was observed to produce statistically significant reductions in dental plaque, measuring 24.4% in the morning pre-brushing, and 27.9% in the afternoon assessment. Tooth brushing was observed to significantly reduce plaque on the teeth in the morning post-brushing for subjects using either the standard NaF or the SnF2 dentifrice. There were no statistically significant differences in plaque coverage on teeth following tooth brushing with the NaF (6.3% coverage) or SnF2 (6.2% coverage) dentifrices. CONCLUSION: Use of a 0.454% SnF2 dentifrice produced statistically significant reductions in dental plaque formation as compared to the similar use of a standard 0.243% NaF dentifrice. PMID- 17410953 TI - Laboratory interproximal access efficacy and gingival margin cleaning of the elmex SENSITIVE SOFT, EXTRA SOFT and ADA reference toothbrushes. AB - OBJECTIVE: The purpose of these laboratory studies was to evaluate three toothbrushes for their ability to remove artificial plaque deposits at interproximal sites and along the gingival margin of simulated anterior and posterior teeth, using horizontal and vertical brushing motions. METHODOLOGY: Three toothbrushes were evaluated: elmex SENSITIVE SOFT; elmex SENSITIVE EXTRA SOFT; and the ADA reference. Twenty-four tests on each toothbrush group were conducted, and results were statistically analyzed using ANOVA and the post hoc Tukey test. RESULTS: In both the interproximal access efficacy and gingival margin cleaning assays, the elmex SENSITIVE EXTRA SOFT and SENSITIVE SOFT toothbrushes were statistically superior (p < 0.001) to the ADA reference toothbrush. CONCLUSION: The elmex SENSITIVE SOFT and EXTRA SOFT toothbrushes are predicted to offer excellent plaque removal efficacy interproximal areas and along the gingival margin of both anterior and posterior teeth. PMID- 17410955 TI - A 4-year-old child with a 3-mm left upper eyelid ptosis. PMID- 17410954 TI - Evidence against mobile pulleys on the rectus muscles and inferior oblique muscle: central nervous system controls ocular kinematics. PMID- 17410956 TI - Diagnosis and treatment of congenital nasolacrimal duct obstruction. PMID- 17410957 TI - What's your diagnosis? Wilson disease (hepatolenticular degeneration) with Keyser Fleischer ring. PMID- 17410958 TI - Management of acute conjunctivitis in school-aged children. PMID- 17410959 TI - Retinal detachments in the pediatric population: part II. PMID- 17410960 TI - The characteristics and surgical outcomes of medial rectus recessions in Graves' ophthalmopathy. AB - PURPOSE: To evaluate the clinical characteristics and surgical outcomes of medial rectus (MR) recessions in patients with Graves' ophthalmopathy. PATIENTS AND METHODS: The clinical records of 32 patients with Graves' ophthalmopathy who underwent MR recessions with adjustable sutures for restrictive esotropia were reviewed. The clinical characteristics of patients, the size of the esodeviations, the limitations of ductions, the surgical doses, and observed responses to surgery were recorded and analyzed. Main outcome measures included the ratio of predicted to observed correction for MR recessions, improvement in ductions, and restoration of binocular status. RESULTS: The mean age of the 32 patients (20 women, 12 men) at surgery was 54.1+/-11.4 years. The mean duration of thyroid eye disease was 4.3+/-5.4 years (range, 1 to 24 years). The ratios of predicted to observed correction for esodeviations at distance and near, respectively, were 2.21+/-1.24 and 2.16+/-1.81 at the time of adjustment and 1.61+/-0.37 and 1.84+/-0.90 at final follow-up. The limitation of abduction improved from -2.3+/-1.3 to -0.75 +/-0.98. Binocular single vision was achieved in 73% of patients, and a further 10% of patients were able to fuse with prisms. A history of decompression was present in 75% of cases. Patients with a history of decompression had more restriction in abduction (-2.49 vs -1.78, P =.061), more frequently required bilateral surgery (75% vs 62.5%), and had a higher ratio of predicted to observed correction (1.71 -0.37 vs 1.37+/-0.28, P = .043). CONCLUSIONS: Patients with Graves' ophthalmopathy who undergo MR recession for restrictive esotropia are prone to undercorrection. A history of decompression is associated with a less favorable clinical outcome. Augmented surgery, adjustable sutures, or both are recommended for improved surgical outcomes. PMID- 17410961 TI - Exclusion of students with conjunctivitis from school: policies of state departments of health. AB - PURPOSE: To use current state department of health regulations regarding exclusion of students with conjunctivitis from school as a starting point in developing uniform recommendations for schools. METHODS: State departments of health were asked to state their policy regarding when a child with conjunctivitis may return to school. This information was collated and examined for trends. The results were compared with current literature on infectious conjunctivitis. RESULTS: Of the 43 states that responded, 7 allow children with conjunctivitis to remain in school, 8 allow their return once antibiotic treatment is initiated, 12 allow their return 24 hours after antibiotics are initiated, 13 exclude them until the disease is noncommunicable, and 16 require the approval of a physician for return to school. Seventeen states gave multiple recommendations, which were often contradictory. CONCLUSIONS: Although no current consensus exists among state health officials regarding students with conjunctivitis, the literature supports excluding children with conjunctivitis from school until they are asymptomatic. When patients are treated with fourth generation fluoroquinolones, the length of exclusion may be as little as 24 hours in cases of bacterial conjunctivitis, and longer in cases of viral conjunctivitis. Following these guidelines may prevent epidemics of bacterial and viral conjunctivitis. PMID- 17410962 TI - Orbital space-occupying lesions in Indian children. AB - PURPOSE: To present a review of all patients younger than 16 years who presented to us with orbital space-occupying lesions. METHODS: Retrospective case study. RESULTS: We analyzed the records of 119 children younger than 16 years of age who had presented to us with proptosis during the 10-year study period. Myogenic tumors were the most common lesion seen (24%), followed by metastatic and secondary orbital tumors (17%), vasculogenic lesions (15%), cystic lesions (10%), inflammatory lesions simulating tumors (7.5%), optic nerve and meningeal tumors (7.5%), peripheral nerve tumors (5%), lacrimal fossa lesions (4%), lymphoid tumors and leukemias (2.5%), primitive neuroectodermal tumors (3%), and others (2.5%). CONCLUSION: We analyzed the profile of orbital lesions that underwent surgical procedures and were proven histopathologically. We found 63 malignancies among the 119 cases, for a malignancy rate of 53%. Orbital rhabdomyosarcoma and retinoblastoma with orbital spread were the most common causes of proptosis among the children. PMID- 17410963 TI - Central corneal thickness in pediatric glaucoma. AB - PURPOSE: To compare the central corneal thickness (CCT) of children with and without glaucoma and to estimate the effect CCT may have on the intraocular pressure (IOP) assessment of children. PATIENTS AND METHODS: CCT and IOP measurements were performed in an unmasked fashion in 34 consecutive children with glaucoma and 28 consecutive children without glaucoma. Patients with corneal edema or extensive corneal scarring were excluded. IOPs in the pediatric glaucoma group were corrected for CCT using a linear algorithm; adjustments of 3 mm Hg or more were considered clinically significant. RESULTS: Mean CCT for eyes with glaucoma was 610.2+/-121.9 microm; for the control eyes, mean CCT was 555.6+/ 38.4 microm (P < .001). Mean CCT was 543.3+/-66.9 microm for eyes with primary congenital glaucoma, 591.9+/-23.1 pm for those with Sturge-Weber syndrome, 662.7+/-68.7 microm for those with aphakic glaucoma, 754.5 +/-92.6 microm for those with aniridia, and 820.6+/-133.7 microm for those with microcornea (P < .001). Applying a correction formula for IOP with CCT adjustment, the adjusted IOP was overestimated by > or =3 mm Hg in 14 (41.2%) eyes and underestimated by > or = 3 mm Hg in 5 (14.7%) eyes. CONCLUSIONS: The mean CCT was higher for the pediatric glaucoma group than for the control group. However, the CCTs varied significantly depending on the specific diagnoses. CCT was estimated to have a clinically significant effect on IOP measurements in more than half of the patients with pediatric glaucoma. Pachymetry results should be considered in the management of these patients. PMID- 17410964 TI - Systemic metastasis following hyphema drainage in an unsuspected retinoblastoma. AB - A 6-year-old girl had total hyphema and elevated left intraocular pressure following trivial trauma. B-scan with vector A-scan revealed vitreous opacities consistent with hemorrhage. The drained hyphema did not recur. A left vascular conjunctival mass and massive cervical lymphadenopathy occurred 7 months later. Biopsy revealed extraocular retinoblastoma and lymph node metastasis. Computed tomography showed an intraocular mass with intracranial extension. She died of metastatic disease despite intensive chemotherapy. Retinoblastoma should be suspected in a child with hyphema following trivial trauma. PMID- 17410965 TI - Rare ocular and systemic associations in a case of neurofibromatosis 2. AB - We describe a child with an extensive medical history who presented with extraocular muscle paresis that, on investigation, led to the diagnosis of neurofibromatosis 2. This child had multiple endocrinologic abnormalities and rare ophthalmologic features that could be associated with the disease. PMID- 17410967 TI - Unilateral neuroretinitis associated with nasal cilioretinal artery occlusion. PMID- 17410966 TI - Rare presentation of fungal orbital cellulitis in an immunocompetent infant. AB - A 3-month-old infant presented with unilateral proptosis. Ophthalmic and physical examinations, computed tomography, and histopathologic examination of a biopsy specimen were performed. Histopathologic examination confirmed fungal orbital cellulitis. The patient responded to 6 weeks of antifungal therapy. Fungal orbital cellulitis should be included in the differential diagnosis for infants with proptosis. PMID- 17410968 TI - [Nutrient content and adequacy of roughage for horse nutrition: an investigation under Swiss conditions]. AB - For horses no special tables related to nutrients for Swiss roughage exist. For this reason samples of hay, straw, silage/haylage and green forage were taken from 46 horse keeping farms in 22 cantons. The samples were judged by sense and following the nutrient--and macromineral--content as well as the content of fructans were analysed. Regarding its quality no sample was totally inadequate for horses. The mean contents of crude protein in Swiss hay for horses were clearly lower than in hay for ruminants and in German hay for horses. The mineral contents (calcium, magnesium, phosphorus) showed a wide range. On average they were also lower than the values provided in tables for ruminants. Except for one sample the silages/haylages showed a dry matter content of more than 40 %. No nutrient tables for silage or haylage, which are such high in dry matter contents, were found in the literature. The contents offructans in silage/haylage and green forage also showed a wide range with a maximum of 94 g/kg DM fructan. PMID- 17410969 TI - Lesion profiles and gliosis in the brainstem of 135 Swiss cows with bovine spongiform encephalopathy (BSE). AB - Lesion profiles are considered to be an important tool for the comparison of the various animal and human spongiform encephalopathies and to obtain information upon prion strain variations. Histological and immunohistochemical reactions (PrPsc, GFAP) in 13 brain areas at 4 levels in the brainstem from 135 BSE positive and 45 BSE-negative cases were retrospectively evaluated. In this retrospective study a lesion profile based on histological features was worked out on the basis of BSE cases originating from Switzerland over a period of ten years. They were confirmed post mortem by histology and immunohistology. Our findings were reviewed in comparison with lesion profiles published in England. No striking differences comparing type and quality of lesions in the relevant areas between the Swiss and the English cases were evident. Moreover, the lesion profiles and the character of the lesions did not differ between animals born before or after the offal feeding ban, which supports the hypothesis that the Swiss epidemic is sustained by the same single, stable strain of the BSE agent, which is probably the same as in the English epidemic. There was a good correlation between PrPsc accumulation and spongiform changes, in particular in those areas which were morphologically most affected. Astrocytosis in BSE was quantified. A significant rise in GFAP-positive cells could be shown comparing the brain stem nuclei of BSE affected with BSE-unaffected cattle, despite considerable variation between the cases and between the nuclei. The observed astrocytosis did correlate with vacuolation of the neuropil and of perikarya as well as with PrPsc accumulation. PMID- 17410970 TI - [A case of alveolar hydatid disease in a dog: domestic animals as rare incidental intermediate hosts for Echinococcus multilocularis]. AB - A 2-years old male Labrador retriever dog was presented with intermittent therapy resistant diarrhoea, accompanied by vomiting, inappetence, apathy, and mild fever. The blood analysis showed an anaemia, neutrophilia, eosinophilia, and increased liver enzymes. Abdominal palpation was slightly painful. X-rays and echography revealed a severely enlarged liver with multiple cavernous structures. Histopathologic examination of liver biopsies showed a severe chronic granulomatous hepatitis with numerous parasitic cysts. Morphology of the cysts was compatible with the metacestode stage of Echinococcus multilocularis. The dog was only 2-years old at the time of diagnosis. Although alveolar hydatid disease of the liver is rare in dogs, it should be envisaged as a possible differential diagnosis in cases of space-occupying processes in the liver, even in young animals, as the incubation period of this disease in the dog can be considerably shorter than in humans. PMID- 17410971 TI - Lawsonia intracellularis proliferative enteropathy in a foal. AB - A weanling foal was diagnosed with proliferative enteropathy caused by Lawsonia intracellularis based on history, clinical findings of depression, anorexia, weight loss, colic, diarrhea, and ventral edema, and a combination of serology and fecal PCR. An epidemiological investigation on the premises revealed that many of the other foals and adult horses were seropositive for L. intracellularis, despite being clinically normal, and identified a dog as a potential carrier and source of infection for the foal. The foal was successfully treated with a combination of azithromycin and rifampin. PMID- 17410972 TI - Should we worry about the enthusiasm toward evidence-based health promotion practices? PMID- 17410973 TI - The Well-being Profile--an Internet tool for school health promotion. AB - For years, the WHO has, in accordance with the 'settings' idea, encouraged a whole school approach when trying to promote health and well-being in schools. This developmental study analyses the implementation and pilot phase experiences of a holistic well-being evaluation tool for schools, the School Well-being Profile, on the Internet. The Profile is based on the theoretically established School Well-being Model. The School Well-being Profile consists of electronic survey forms and an automatic facility that analyses and produces results on the data in graphic and numeric form. After the data has been entered, the primary user within the school can immediately view and print out the results. The figures can be compared with the averages of all schools to pinpoint areas where well-being is different from that in the other schools. The Profile resides on a Finnish National Board of Education server (www2.edu.fi/hyvinvointiprofiili) and its use is free of charge for all schools. The Profile became popular in its first year: it was used by 33 primary schools, 28 lower secondary schools and 9 upper secondary schools with a total of 9,169 respondents. Overall, 94% of the students and 99% of the personnel expressed that it had been at least fairly easy to fill in the questionnaire. The paper shows that theoretical research in health promotion can effectively be put into practice using information technology tools. The project that produced the School Well-being Profile ended in 2004, yet the WWW-Profile continues to gain new users. The school administrators, personnel and students have found the Profile easy to use and worth the time and effort invested by the school. The strategies adopted in the design and dissemination of the Profile seem to have been successful in providing a sustainable resource to contribute to the future promotion of well-being in schools. PMID- 17410974 TI - Maintaining population health in a period of welfare state decline: political economy as the missing dimension in health promotion theory and practice. AB - There is increasing recognition in the health promotion and population health fields that the primary determinants of health lay outside the health care and behavioural risk arenas. Many of these factors involve public policy decisions made by governments that influence the distribution of income, degree of social security, and quality and availability of education, food, and housing, among others. These non-medical and non-lifestyle factors have come to be known as the social determinants of health. In many nations--and this is especially the case in North America--recent policy decisions are undermining these social determinants of health. A political economy analysis of the forces supporting as well as threatening the welfare state is offered as a means of both understanding these policy decisions and advancing the health promotion and population health agendas. The building blocks of social democracies--the political systems that seem most amenable to securing the social determinants of health--are identified as key to promoting health. Health promoters and population health researchers need to "get political" and recognize the importance of political and social action in support of health. PMID- 17410975 TI - Revitalizing health promotion: a case study from Serbia. AB - The last two decades of the twentieth century were difficult ones for health promotion in Serbia. The political, economic and social upheaval resulting from the civil conflict in the former Republic of Yugoslavia put enormous pressure on the health system overall and undermined a previously strong and effective health education and promotion service. In recent years the government, supported by external donors, has refocused attention on the need for high quality public health, including preventive activities as a central goal of the health reform process. This process is reviewed through a case study of a high profile health promotion campaign recently completed within the country. The potential for Serbia to regain its position as an innovator in community-based health care as well as the obstacles to be overcome are analysed. PMID- 17410976 TI - Bloomberg Global Initiative to Reduce Tobacco Use: the International Union for Health Promotion and Education partners for the establishment of a global clearinghouse for effective anti-tobacco advertising. PMID- 17410977 TI - [Understanding local concepts of equity to formulate public health policies in Burkina Faso]. AB - Equity is an essential health promotion concept and must be included at the heart of public health policy making. However, equity, which can also be referred to as social justice, is a polysemic and contextual term which definition must stem from the discourse and values of the society where the policies are implemented. Using a case study from Burkina Faso, we try to show that the non-acknowledgement of the local concept of social justice in the policy making process partly explains the resulting policies' relative failure to achieve social justice. Data collection methods vary (individual and group interviews, concept mapping, participant observation, document analyses) and there are qualitative and quantitative analyses. The four groups of actors who generally participate in the policy making process participated in the data collection. With no intention to generalise the results to the entire country, the results show that mass social mobilisation for justice is egalitarian in type. Health or social inequalities are understood by individuals as facts which we cannot act upon, while the inequalities to access care are qualified as unjust, and it is possible to intervene to reduce them if incentive measures to this effect are taken. We also observed a certain social difficulty to conceive sub-groups of population and fierce will to not destabilise social peace, which can be provoked when looking for justice for the impoverished sectors of the population. This research allows better understanding about the emic aspect of equity and seems to confirm the importance of taking into account local values, especially social justice, when determining public policy. PMID- 17410978 TI - [The usefulness of pelvic lymphadenectomy in bladder cancer]. AB - OBJECTIVE: [corrected] To assess the usefulness of pelvic lymphadenectomy in bladder cancer. METHODS AND RESULTS: With the followings key words: "bladder cancer, lymphadenectomy, lymph node metastasis" we search in Medline/PubMed database for papers published during the last ten years. Thirty three papers fulfilling the search criteria were selected. CONCLUSIONS: It does not exist any randomized prospective study comparing different pelvic lymphadenectomy techniques. Nodal metastasis in bladder cancer after radical cystectomy and pelvic lymphadenectomy ranged between 18% and 28%. Standard lymphadenectomy could improve tumor staging and probably survival in selected patients. It is advisable to remove, as a quality parameter, at least 10 to 14 nodes in a pelvic lymphadenectomy. Lymph node density seems to predict survival better than TNM staging system. It is advisable to perform separate lymph node dissection rather than en-bloc. PMID- 17410979 TI - [Penis epidermoid tumors review]. AB - OBJECTIVES: To study the characteristics and evolution of the epidermoid penis tumours. MATERIAL AND METHODS: It was carried out a retrospective study on the epidermoid penis tumours treated in our center between 1981 and 2005. RESULTS: 16 tumours penis epidermoides were diagnosed. The average age of the patients was of 71.7 years (interval between 54 and 90 years). In 80% of the cases they are diagnosed in advanced stadiums (T3 and T4). The most habitual presentation forms are the ulcerous lesions (53%) and papilar (33%). The average diameter of the lesion was 2.3 cm, and the most frequent localization the glands (53%) and balano prepucial (33%). They were carried out 7 local scissions, 8 partial penectomies, one total penectomy, and an local scission pluslocal treatment with 5-fluoracile. With a follow-up of 24 months 5 relapses took place, mainly in patients subjected to local excision of the lesion (p = 0.06 log-rank test). CONCLUSIONS: The epidermoid carcinoma of the penis is a tumoral lesion characteristic of advanced ages that is diagnosed later and treated in not very aggressive way. Therefore it is frequent the advanced stadiums and the tumour relapses. PMID- 17410980 TI - [Yield diagnosis of the peripheral cores in 10 needle extended prostate biopsy]. AB - OBJECTIVE: To value if the increase in the number of cylinders in the prostate's biopsy raise the diagnostic performance of this test. MATERIAL AND METHOD: In March 2005 we initiate this prospective study with patients who are subject to a first prostate Biopsy by a PSA between 4 and 10 ng/ml and negative rectal touch. Transrectal, echodirected prostatic biopsies with ten punctures are carried out according to the following programme as follows: 6 Biopsies according to classic sextant technique, to what we add 4 cylinders from the most lateral zones of the prostate (lateral horns). The variables are analyzed: Age, Total PSA, Free PSA/Total PSA, prostatic volume and PSA density. RESULTS: We value 90 patients consecutively. Prostatic adenocarcinoma was diagnosed to 37 of the 90 patients, which means that the diagnostic rate of the extended Biopsy reached a 41%. Nevertheless, the rate for the classic sextant Biopsy was 32.3%. These differences are statistically significant (Mcnemar test 0.008); besides, this supposes a 27% increase in the diagnostic rentability. The "extra" cylinders in the lateral horns detected 8 tumours which were not detected in the cylinders of the the sextant, and 5 of them presented a Gleason higher or equal to 6. CONCLUSIONS: In our centre we think that the Biopsy extended to 10 cylinders is an adequate strategy for potential prostatic adenocarcinoma patients who are going to be subject to their first Biopsy. Realizing the clinic relevance of the tumours diagnosed thanks to "extra" cylinders, we do not think that this modality implies an over-diagnosis and consequently an overtreatment of the prostate cancer. PMID- 17410981 TI - [Laparoscopic augmentation enterocystoplasty: initial experience]. AB - OBJECTIVE: To analyze the initial experience of our group in the realization of the augmentation enterocystoplasty by laparoscopyc approach. METHODS AND PATIENTS: We describe the augmentation enterocystoplasty technique with ileal segment completely achieved by laparoscopyc approach. We present the cases of two patients suffering from hyperreflexic bladder refractory to medical treatment who underwent this surgery. In both cases the technique was realized without intraoperative complications although it was needed a surgical time of 6 and 4.5 hours respectively. The results after 12 and 5 months were satisfactory in both patients, obtaining a low pressure bladder with a good continence. CONCLUSIONS: Laparoscopyc augmentation enterocystoplasty is a complicated technique that requires a great experience, mainly in laparoscopyc suture. It reproduces completely the open surgery and it offers all the advantages inherent to the laparoscopyc surgery. PMID- 17410982 TI - [Bladder neoplasm after nephroureterectomy for upper urinary tract tumor. Does distal ureter surgery influx on the evolution?]. AB - OBJECTIVE: To compare evolution in patients with urothelial upper tract tumor (UUTT) in whom we performed the classic open approach to the distal ureter with those in whom whe performed the endoscopic approach. We study the bladder recurrences in each group and the factors which may influx on the evolution. MATERIAL AND METHODS: Retrospective review of nephroureterectomies in our department in the last 20 years. Group A (n:24): double incision (lumbar and pelvic incision), Group B (n:29): endoscopical approach of the distal ureter and classic lumbar incision. A descriptive study is first performed in which we review: sex, and patients age, background transuretral resection of bladder tumor (TURBT), UUTT side, and UUTT location (calices, pelvis or urether). Pathological stage and tumor grade are also exposed. Secondly, a review of the bladder recurrences in each group is performed. In order to find differences between de groups we used the Fisher's Exact test. RESULTS: We observed that in terms of bladder recurrence there is a statistically significant difference between the two groups favoring Group B (p < 0.036), which means that there are less bladder recidives when perform endoscopic approach of distal ureter. Noneless due to the groups inhomogeneity in tumor stage, grade, and location, this differences seem to be associated to these sigues, more than to the approach to the ureter as an independant variable. In that concerning to the eventual relationship between bladder recidive and background of previous TURBT, we have not found any differences between the two groups but there are statystically significant difference in the global series. The small number of recurrences (13) does not allow us to establish a well based conclusion on this issue, but it seems that the background of previous TURBT is an important factor that may influx in posterior bladder recurrences. CONCLUSIONS: In our results, endoscopic approach of distal ureter in nephroureterectomy for UUTT does seems to positively influx in the posterior chance of bladder recurrences. But the studied sample is small and the two groups have not been aleatorized because of ethical reasons. May the low number of cases affect the test's statistical potency. It seems that previous TURBT may conditionate these recurrences, as we show in global serie. It should be necessary a prospective and statistically analised multicenter trial to understand if distal ureter approach influx on the evolution of these tumors. PMID- 17410983 TI - [Assesmment urethrorrhagia in childhood]. AB - OBJECTIVE: [corrected] To review urethrorrhagia cases, the need of complementary test, its management and natural history. MATERIAL AND METHOD: A retrospective analysis was carried out for the 12 patients who suffered from urethrorrhagia, consecutively diagnosed in our department. Parameters as age, and micturitional habit were evaluated. In this way, we described the diagnostic techniques used, assessment and the follow-up of each patient. RESULTS: All cases correspond to male children with an average age of 8.7 years (range: 18 months-14 years). Urethrorrhagia appears in 100% of the patients, episodes which were daily in 58.4%. 33% (4/12) of them also present other micturitional symptoms. 75% (9/12) had a lazy micturitional habit. Among the urological backgrounds we found: 1 patient was circumcised 3 years ago, a meatotomy was done in other due to meatus stenosis 2 years before, and a last one had an electrocoagulation of a urethral polyp just 1 month before. 83.4% (10/12) of urine cultures were negative. An urethrocystoscope was done in 9 patients finding a verum hypertrophy in 4, inflammatory changes next to membranous urethra in 3, scar at the bulbar urethra in 1 and in a last one no abnormalities were found. Out of 8 patients with lazy micturitional habit, 75% (6/8) resolve spontaneously once they correct their habits after a mean follow-up of 9 months. Only 2 patients needed surgery (TUR). In the other 2 patient, symptoms. CONCLUSIONS: Urethrorrhagia in childhood is a benign condition in most cases, which is almost always cured spontaneously during the first 2 years of follow-up. Radiological studies as well as endoscopic procedures are unnecessary in the early management of these patients thus being relegated to recurrent or persistent bleeding. PMID- 17410984 TI - [Intratesticular calcifications in childhood]. AB - INTRODUCTION: Testicular microlithiasis (TM) represents an uncommon occurrence at paediatrics with a radiological incidence between 0.16 to 0.6% according to reports. The greater use of ultrasound as well as the availability of high frequency probes has increased the number of incidental TM being diagnosed. MATERIAL AND METHOD: From January 1998 to October 2002 seven children, aged 9 to 13 years (average 11 years and 1 months), were diagnosed of MT in our department. Sonographic appearance was hyperechogenic multiple small foci of 1-3 mm without acoustic shadowing of the testicular parenchyma. AIM: To evaluate sonographic patterns of TM, their associations, clinical consequences and to determine their management. RESULTS: All children were asymptomatic and the reasons for the ultrasound were: cryptorchism (3), short height (1), gynecomastia (1), varicocele (1) and scrotum trauma (1). It should be noted that 3 of the cases showed bilateral TM. Out of 10 testicles with TM, 8 met classic testicular microlithiasis (CTM) criteria (at least one image that showed five or more microliths in either or both testes) and 2 had limited testicular microlithiasis (LTM) (to have at least one microlith). Clinical and radiological follow-up is being performed annually, ranging from 18 months to 6 years, during which there hasn't been progression or regression in the number of TM or in their distribution. In none of these cases there has been shown a related testicular cancer. CONCLUSIONS: In other broad reviews it has been shown that there is no evidence to suggest that the TM have to be considered as premalignant lesions by themselves. However, the association among TM and cancer exists. Because of that, and due to the lack of consensus, we recommend annual clinical and radiological (ultrasound) follow-up until puberty, and thereafter the patient should be informed of unknown natural history of this condition. We do not recommend more invasive procedures such as biopsy except: 1) apart form the TM there is a change in the echogenicity or 2) if there is a change in the number or distribution of the TM. PMID- 17410985 TI - [Usefulness of renal resistive index in the diagnosis and evolution of the obstructive uropathy. Experimental study]. AB - OBJECTIVE: The study was focused on determining the effectiveness and reliability of using the renal resistance index as a method of diagnosing and monitoring the evolution of obstructive uropathy treatment. For this purpose, we conducted an experimental study on a homogenous group of animals that all had the same level of obstruction. MATERIAL AND METHODS: 15 healthy female pigs were used. The experiment was divided into three phases: phase I consisted of a study prior to unilateral pyeloureteral junction obstruction, performing retrograde ureteropyelography, renal B-mode ultrasound and duplex-Doppler (of both kidneys) at a level of the arcuate arteries. Then, the obstruction was performed on the animals. Phase II commenced by diagnosing the lesion, 6 weeks after the previous phase, by means of the aforementioned diagnostic methods. Finally, the endourological treatment was completed. Animals were monitored (Phase III) 15 weeks after the endopyelotomy, using the same methods as in the study, by assessing the urinary tract (fluoroscopy) and both kidneys by determining the renal resistance index and ratio. RESULTS: All the animals in the study showed signs of urinary obstruction on radiology and renal ultrasound 6 weeks after ureteral ligature. After treatment and follow-up, all animals showed signs of having recovered from the obstructive uropathy. Values of deltaRI during the 3 phases are detailed below. Fase I deltaIR = 0.01, Fase II deltaIR = 0.11, Fase III deltaIR = 0.02. CONCLUSION: RI determination using duplex-Doppler is effective for distinguishing obstructive dilatation using non-invasive techniques. However, this parameter provides very slight differences and it can also be influenced by too many direct and indirect factors (observational, patient age, anaesthesia, haemodynamic parameters, etc.), to supplant the classic diagnostic methods. PMID- 17410986 TI - [Low obstruction tract by system Reemex]. AB - The Remeex system it's a sling with regulable tension created for the surgical correction of the female urinary incontinente. The system contains a varitensor central part of the protesis that regulates the tension of the uretra. The screwdriver that acts against the varitensor to the modify the tension under the uretra in the postop inmediately or time after also. We present a clinic case of a patient with diagnostic of urinary stress incontinence and the surgical treatment with advantages against others kind of systems used for incontinence treatment at this time. PMID- 17410987 TI - [Torsion of undescended testis]. AB - Torsion of a cryptorchidid testicle presents a non-specific symptomatology. Clinical suspicion indicates surgical exploration, irrespective of Doppler ultrasound with its inherent false negative results. With regards to treatment, an orchiectomy is performed when the testis is necrotic. Management of the contralateral testis is controversial. Our policy is not to fix them systematically because we believe that a healthy education of parents and children is more important, so as to obtain an earlier diagnosis of a future torsion. PMID- 17410988 TI - [Acute myocardial infarction associated to the Sildenafil consumption. A case report and review of the literature]. AB - Erectile dysfunction affects more than 30 million men in The United States. Since the FDA approved the use of Sildenafil, prescription of this medication has been raising. Adverse events of Sildenafil includes: fatigue, dyspnea, and hypotension. Reported adverse cardiac events associated with the medication use include myocardial infarction, ventricular tachycardia, angina and death, raising concerns about the safety of this agent in patients with coronary artery disease. Published guidelines regarding the management of cardiac patients with erectile dysfunction suggest that Sildenafil may be hazardous in patients with ischemic heart disease. In patients using Sildenafil, myocardial infarctions have been reported to the Food and Drug Administration. Now, we report a patient with myocardial infarction after taking 100 mg of Sildenafil without sexual activity. PMID- 17410989 TI - [Bilateral and synchronic testicular neoplasia. A report of a new case]. AB - Bilateral and synchronic testicular tumor is an unfrequent fact (0,17% of germinative tumors of testicle), being most cases seminomas. We present the case of a male, 25-year old patient underwent a bilateral radical orchiectomy for bilateral and synchronic testicular tumor. The result of anatomopathological examination revealed bilateral seminoma (pT2 in both testes). Patient received four courses of adjuvant chemotherapy (etoposide and cisplatin) for metastases (N2, M1). The patient is without evidence of disease two years after orchiectomy. PMID- 17410990 TI - [Parameatal urethral cysts. 3 new paediatric cases and literature review]. AB - We present 3 new cases of urethral parameatal cysts in paediatric age. Literature is reviewed and histological data and ambulatory management are discussed. PMID- 17410991 TI - [Multiple bladder stones]. PMID- 17410992 TI - [Late complications of a ureterosigmoidostomy]. PMID- 17410993 TI - [Complex renal cyst (renal pseudocyst)]. PMID- 17410994 TI - [Right renal cell carcinoma with inferior vena cava thrombous]. PMID- 17410995 TI - What is not part of the child abduction news story? PMID- 17410996 TI - Problem behaviors in adolescents. AB - A fifth or more of a nationally representative sample of school-attending adolescents report engaging in problem behaviors such as skipping school, using alcohol, fighting, shoplifting, and stealing. A smaller but significant number of adolescents report engaging in risky sexual behaviors. All of these behaviors have potentially serious consequences for adolescents, their family and friends, their school, and society. Nurses are in a unique position to help identify these behaviors in adolescents, educate school personnel about the behaviors, educate adolescents about the risks they face when they engage in these behaviors, and assist parents to access the resources they need to help children who may be engaging in these problem behaviors. PMID- 17410997 TI - Parenting styles and treatment of adolescents with obesity. AB - Professional caregivers have an important task in building a trusting relationship with parents and adolescents and in supporting parents in their parental roles. Our clinical experience of some 300 adolescents with obesity between 9 and 18 years of age and their parents has convinced us that consideration of parenting styles is fundamental in the treatment of children and adolescents with obesity. Typical case situations supporting the significance of parenting styles and illustrating the relationships between parents and adolescents with obesity can be identified. Group sessions with parents are the preferred mode for discussing typical parenting issues in the management of obese adolescents. The purpose of this paper is to describe different parenting styles, and to present a set of typical case situations and treatment strategies for nurses working with adolescents with obesity. PMID- 17410998 TI - Evaluation of a PNP care coordinator model for hospitalized children, adolescents, and young adults with cystic fibrosis. AB - PURPOSE: The purpose of this study was to evaluate a new model for delivery of care to hospitalized children, adolescents, and young adults with cystic fibrosis (CF) admitted to the adolescent unit at Golisano's Children's Hospital at Strong. SPECIFIC AIM/HYPOTHESIS: The specific aim of this study was to test the hypothesis that children, adolescents, and young adults with CF who were hospitalized under the new model for care delivery would have better experiences and clinical outcomes than they had during previous hospitalizations prior to the implementation of the new model. METHODS: A pre-post design included all children, adolescents, and young adults with CF admitted to the hospital for treatment of a pulmonary exacerbation during the six-month study period. Each patient's experience was compared with his or her most recent previous hospitalization. Outcomes measured included timeliness of inpatient consultations, change in forced experitory volume in first second (FEV1), weight gain, length of hospital stay (LOS), and patient/parent and nursing/medical staff satisfaction. RESULTS: A total of 21 patients completed the study. Compared to the patient's previous admission, there was a significant reduction in the time to complete consultations by ancillary services. The differences between predicted LOS and actual LOS was reduced by 2.47 days (p = .06). Actual LOS was decreased by 1.35 days. Parent/patient satisfaction with new model remained high and health care provider satisfaction was overwhelmingly positive. CONCLUSIONS: An inpatient Pediatric Nurse Practitioner Care Coordinator (PNP-CC) for CF patients admitted to the hospital can reduce the time for ancillary service consultations, reduce LOS, and improve patient and health care provider satisfaction. This model also can be applied to the treatment of hospitalized children with other chronic illnesses. PMID- 17410999 TI - Asthma care of children in clinical practice: do parents report receiving appropriate education? AB - This paper describes asthma education received from the health care provider as reported by parents of children with asthma and evaluates differences in their report based on socio-demographic and disease characteristics. METHODS: Parents of 228 children with asthma were recruited from diverse clinical practice sites and asked to report on the level of education received in key content areas the National Asthma Education and Prevention Program identified as critical. RESULTS: The overall proportion of education content fully discussed was 66%. Areas that dealt with acute management (how to manage an asthma attack: 75%) and medication administration (how to use an MDI: 81%) were most likely to be discussed. Content dealing with ongoing chronic management and collaborative care planning (discussing goals of management: 44%; providing written guidelines for acute management: 44%) was significantly less likely to be discussed. Education reported differed significantly based on symptom severity and socio-demographic characteristics. Parents of children with moderate to severe persistent symptoms reported receiving more education than those of children with mild symptoms (p < .05). Minority and poor parents, parents with less education, and parents whose children received health care in a clinic practice setting reported more education received than did their counterparts (p < .05). SUMMARY: Significant gaps exist in education received overall, and specifically in areas associated with developing a collaborative relationship between parent and health care provider. The initial observation of socio-demographic differences in education reported has implications for pediatric nurses involved in asthma education and warrants further investigation. PMID- 17411000 TI - Down on the farm: preventing farm accidents in children. AB - Agriculture continues to rank as the most dangerous industry in the United States. The objectives of this pilot study were to identify the incidence of care provided to children involved in farm accidents, examine barriers to utilization of farm safety materiaJs by primary health care practitioners (PHCPs), and determine the percentage of PHCPs who provide anticipatory guidance related to farm safety. Pender's revised Health Promotion Model (2006) was used. A survey was developed and mailed to 110 PHCPs of all levels in North West Pennsylvania. The return rate was 20%. Types of injuries identified by the PHCPs included lacerations and musculosketal injuries caused by animals and farm equipment. Forty-five percent of the PHCPs reporting asked new patients/families if their children lived or worked on a farm. Eighteen percent specifically focused on farm related injury prevention. Fourteen percent were aware of farm injury prevention materials and 73% were interested in receiving such materials. A larger study needs to be conducted to validate these findings. PMID- 17411001 TI - The many roles of family members in "family-centered care"--part II. Interview by Deborah Dokken. AB - This article highlights the role parents can play as advocates for their own children. A mother who is interviewed describes a nurse's support in helping her claim her baby as her own in the midst of the NICU environment and how that claiming led her to become involved in his care and a strong advocate for his needs. She shares advice for parents and health care providers. A physician who is interviewed discusses working with this mother and other parents, and describes ways health care professionals can support parents as advocates. PMID- 17411002 TI - Young patients in a young nation: scarlet fever in early nineteenth century rural New England. AB - Children in the United States have benefited considerably from advancements in medical and nursing science over the course of the past 200 years. The twentieth century saw dramatic declines in the incidence of childhood diseases; the prevalence of measles, haemophilus influenzae type B, diphtheria, rubella and tetanus are at all time lows (CDC, 2006). Indeed, many pediatric nurses have never seen any of these diseases, something that would certainly have startled their predecessors just a few generations ago. Before the mid- twentieth century, caring for children with communicable diseases represented the cornerstone of pediatric nursing practice. Now that the incidence has decreased among American children, it is easy to forget about these diseases that once decimated whole communities. This article tries to peel back the mists of history by studying children's health in one rural New England town during the days of the early republic in the 1830s. PMID- 17411003 TI - Complications of diabetes insipidus: the significance headache. PMID- 17411004 TI - Enhancing self-management in children with sickle cell disease through playing a CD-ROM educational game: a pilot study. AB - PURPOSE: To determine whether playing a simple CD-ROM educational game (developed specifically for children with sickle cell disease), improved children's knowledge and confidence in selected symptom management and practice. METHOD: Twenty-two eligible children completed a pretest to determine knowledge and confidence levels, played the Sickle Cell Slime-O-Rama Game, then completed an identical posttest. FINDINGS: Significant increases in knowledge (t = 2.828, p =.010) and confidence (t = 3.759, p =.001) levels between pre- and posttests were identified. CONCLUSIONS: It is promising that a simple, interacting CD-ROM game allowed children with sickle cell disease to quickly acquire knowledge about the disease and symptom management, and increased their confidence to apply this new knowledge. Results suggest the high utility of this tailored game to foster active self-management behaviors in this population. PMID- 17411005 TI - Prescribing medications in pediatrics: concerns regarding FDA approval and pharmacokinetics. AB - Prescribing medications "off-label" is a common practice in pediatric health care since many medications lack U.S. Food and Drug Administration (FDA) approval for pediatric drug labeling due to insufficient drug testing in children. This clinical paper reviews the FDA laws regarding approval of medications in children and the pharmacokinetic differences in absorption, distribution, metabolism and excretion between children and adults. Two commonly used pharmacology resources were reviewed to determine their identification of FDA approved indications in children and dosing recommendations by age or weight in children. Adhering to the "community's standard of care" is a common guideline for prescribing "off-label" in pediatrics, but must be used in combination with multiple respected pediatric resources and with full knowledge of pharmacokinetics in children, particularly in young children. PMID- 17411006 TI - What's new with flu: understanding influenza. PMID- 17411007 TI - Experimental and theoretical investigations on the adsorption of 2' deoxyguanosine oxidation products at oxidized boron-doped diamond electrodes. AB - Electrochemical oxidation of 2'-deoxyguanosine has been performed on boron-doped diamond (BDD) electrodes, resulting in a strong adsorption of the formed oxidized products onto the BDD surface. The adsorption behavior has been investigated by studying the electrochemical behavior of a redox probe ([IrCl6]3-) using cyclic voltammetry. The most probable situations are the formation of (A) an insulating adsorbed film resulting in a partially blocked electrode behavior, (B) a porous film, or (C) an overall conductive film. Different parameters such as the standard rate constant, the charge-transfer coefficient, the electrode/adsorbed products/solution interface resistance, and the formal potential of the redox couple were determined. Through comparison of theoretical current-potential curves obtained by analytical calculations with experimental cyclic voltammograms, we found that the oxidized products of 2'-deoxyguanosine form a continuous conductive film on BDD. PMID- 17411008 TI - Glass nanopore-based ion-selective electrodes. AB - Glass nanopore-based all-solid-state ion-selective electrodes (ISEs) have been developed to probe the distribution of ionic species at micro- or submicrometer length scales, e.g., mapping of ion flux through micrometer-sized pores. The all solid-state ISE was fabricated by sealing a conically etched platinum wire (d = 25-microm; radius of etched tip <10 nm) into a soda lime glass capillary. A Pt disk was exposed by gentle polishing the glass and the disk etched to form a conical pore of submicrometer dimension (radius < approximately 500 nm; depth < approximately 30 microm). Ag was electroplated on the Pt electrode in the pore and gently chloridated to obtain a AgCl/Ag layer within the pore. The AgCl/Ag layer-coated ISE was used as a highly selective Cl- probe in scanning electrochemical microscope experiments to map the ion flux through a micropore. The same ISE was also used as the base transducer of the neutral carrier-doped solvent polymeric membrane. The optimized polymer membranes used for the glass nanopore-based all-solid-state ISE require a higher ratio of plasticizer/polymer (9/1) compared to those for conventional ISE (2/1). An ISE based on deposition of an IrO2 layer at the base of a glass nanopore electrode exhibited a highly sensitive response (79.7 +/- 2.3 mV/pH) to variations in pH and could be used for approximately 3 weeks. PMID- 17411009 TI - Experimentally determining the iR drop in solution at carbon fiber microelectrodes with current interruption and application to single-cell electroporation. AB - Single-cell electroporation uses microelectrodes, capillaries, or micropipets positioned near single, adherent cells to increase transiently the membrane permeability of the cell. The increased permeability permits, for example, transfection without chemical reagents. When using microelectrodes to apply an electric field to the cell, there is a question of how much voltage to apply. Unlike in bulk electroporation, where hundreds of volts may be applied between electrodes, a rather small voltage is applied to a microelectrode in single-cell electroporation. In the single-cell experiment with microelectrodes, a substantial fraction of the voltage is lost at the interface and does not therefore exist in solution. This problem is the same as the classical electrochemist's problem of knowing the "iR" drop in solution and correcting for it to obtain true interfacial potential differences. Therefore, we have used current interruption to determine the iR drop in solution near microcylinder electrodes. As the field is inhomogeneous, calculations are required to understand the field distribution. Results of the current interruption are validated by comparing two independent measurements of the resistance in solution: one value results from the measured iR drop in conjunction with the known applied current. The other value results from a measured solution conductivity and a computed cell constant. We find substantial agreement in the range of resistances from about 2 to 50 kOmega, but not at higher resistances. We propose a simple, four-step plan that takes a few minutes to calculate the approximate current required to electroporate a cell with an electrode of a particular size, shape, and distance from the cell. We validate the approach with electroporation of single A549 cells. PMID- 17411010 TI - Ultra-sensitive quantification of corticosteroids in plasma samples using selective solid-phase extraction and reversed-phase capillary high-performance liquid chromatography/tandem mass spectrometry. AB - Low-dose corticosteroids may provide a favorable benefit/risk ratio for many therapeutic applications. However, the extremely low plasma drug concentrations achieved, in conjunction with the insufficient sensitivity/ selectivity of current analytical methods, renders the evaluation of corticosteroid pharmacokinetics (PK) a significant challenge under such conditions. Furthermore, targeted therapeutic strategies involving administration by inhalation or intraocular injection could result in very low but sustained systemic corticosteroid concentrations, which must be quantified to determine potential side effects. Here we describe a robust method for the ultrasensitive quantification of corticosteroids in plasma samples. This was achieved by the combination of a selective solid-phase extraction (SPE) with a highly sensitive capillary LC (microLC)-MS/MS analysis. SPE washing and elution conditions were optimized so that target drugs are selectively extracted from plasma. By eliminating most undesirable compounds from the sample matrix, this selective SPE procedure enabled a high sample loading volume on the microLC column without compromising chromatographic performance and operational robustness and helped to achieve ultralow detection limits for the corticosteroids in plasma. The effect of microLC separation on the signal-to-noise ratio of corticosteroid peaks in plasma samples was investigated. It was found that with sufficient microLC separation, sensitivity was improved because of a decrease in matrix effects and the removal of endogenous interferences. Detection limits of four clinically important corticosteroids (budesonide, dexamethasone, triamcinolone acetonide, and dexamethasone acetate) ranged from 0.2 to 1 pg/mL in plasma, and linearity was good for all drugs in the range of 5-5000 pg/mL. Accuracy was 88-107% and the variation (CV%) was 2.3-11.1%. A limit of quantification (LOQ) of 5 pg/mL was validated for all four compounds. We applied this method to quantify the low levels of triamcinolone acetonide (TACA) in porcine plasma following suprachoroidal administration, which is necessary to estimate systemic drug exposure resulting from this novel clinical approach for treating inflammatory diseases of the eye. TACA in plasma could be quantified at low pg/mL levels for up to 90 days posttreatment. To our knowledge, this is the first practical analytical approach that can monitor plasma corticosteroids after intraocular administration, given the ultralow plasma concentrations achieved. In summary, this strategy enables PK analysis of corticosteroids in treatment regimens that result in extremely low systemic concentrations, and the approach can be extended for the sensitive quantification of other drugs. PMID- 17411011 TI - Glyphosate and AMPA analysis in sewage sludge by LC-ESI-MS/MS after FMOC derivatization on strong anion-exchange resin as solid support. AB - An innovative analytical method has been developed for the determination of glyphosate and aminomethylphosphonic acid (AMPA), its major metabolite, in sewage sludge. This method involved an alkaline extraction from sludge and purification on a strong anion-exchange resin. While the analytes remained fixed by ionic interactions, an "on-solid support" derivatization by FMOC-Cl was carried out. This versatile approach allowed a 10 min reaction and simple elimination of the excess of reagent. The resulting derivatives remained retained by ionic and hydrophobic interactions with the resin until being eluted by a mixed NaCl water/acetonitrile, 70/30, v/v, solution. After an appropriate dilution and adjustment of the pH, the sample was concentrated on an Oasis HLB solid-phase cartridge. For quality analysis of traces in complex matrixes, LC-ESI-MS/MS in the multiple reaction monitoring positive mode was used fulfilling the European Union requirements (Decision 2002/657/CE). To overcome the matrix effects, stable isotopically labeled standards were added to the sludge extracts as internal standards and were thus derivatized during the procedure in parallel to the analytes. Mean recovery values were 70% +/- 7% for glyphosate and 63% +/- 3% for AMPA. Limits of detection (20 and 30 ppb dw) and limits of quantification (35 and 50 ppb dw) for glyphosate and AMPA, respectively, were sufficient to monitor samples taken from Ile-de-France wastewater treatment plants where contamination currently reached 0.1-3 ppm and 2-35 ppm dw for glyphosate and AMPA, respectively. PMID- 17411012 TI - Graphitized carbon LC-MS characterization of the chondroitin sulfate oligosaccharides of aggrecan. AB - A novel in-gel endoglycosidase technique to study oligosaccharides with graphitized carbon LC-MS has revealed differences in the sulfation profile between the linkage and repeat regions of chondroitin sulfate on aggrecan. Bovine articular cartilage aggrecan was isolated in a composite agarose PAGE gel or diluted in ammonium acetate buffer and was digested overnight with chondroitinase ABC. Including a chemical release/reduction protocol after digestion, we could separate and detect three differentially sulfated chondroitin sulfate disaccharides of the repeat region (DeltaUA1-3GalNAc0/4/6S-ol) from the three differentially sulfated linkage region hexasaccharides (DeltaUA1 3GalNAc0/4/6Sbeta1-4GlcAbeta1-3Galbeta1-3Galbeta1-4Xylitol). Graphitized carbon LC-MS in the negative ion mode was able to resolve isomeric disaccharides and linkage region hexasaccharides. Specific MS2 and MS3 enabled us to confirm the sulfate location on all oligosaccharides by comparing their fragmentation with sulfated disaccharide standards. The presence of unsulfated, 6-sulfated, and 4 sulfated linkage regions was correlated with positive Western blot staining with the respective CS linkage region neoepitope antibodies (1B5, 3B3, 2B6) on digested aggrecan. Our strategy of examining linkage region and repeat region profiles is applicable to screening GAGs from various biological samples in order to detect differences between normal and disease states. PMID- 17411013 TI - Anion and cation mixed-bed ion exchange for enhanced multidimensional separations of peptides and phosphopeptides. AB - Shotgun proteomics typically uses multidimensional LC/MS/MS analysis of enzymatically digested proteins, where strong cation-exchange (SCX) and reversed phase (RP) separations are coupled to increase the separation power and dynamic range of analysis. Here we report an on-line multidimensional LC method using an anion- and cation-exchange mixed bed for the first separation dimension. The mixed-bed ion-exchange resin improved peptide recovery over SCX resins alone and showed better orthogonality to RP separations in two-dimensional separations. The Donnan effect, which was enhanced by the introduction of fixed opposite charges in one column, is proposed as the mechanism responsible for improved peptide recovery by producing higher fluxes of salt cations and lower populations of salt anions proximal to the SCX phase. An increase in orthogonality was achieved by a combination of increased retention for acidic peptides and moderately reduced retention of neutral to basic peptides by the added anion-exchange resin. The combination of these effects led to approximately 100% increase in the number of identified peptides from an analysis of a tryptic digest of a yeast whole cell lysate. The application of the method to phosphopeptide-enriched samples increased by 94% phosphopeptide identifications over SCX alone. The lower pKa of phosphopeptides led to specific enrichment in a single salt step resolving acidic phosphopeptides from other phospho- and non-phosphopeptides. Unlike previous methods that use anion exchange to alter selectivity or enrich phosphopeptides, the proposed format is unique in that it works with typical acidic buffer systems used in electrospray ionization, making it feasible for online multidimensional LC/MS/MS applications. PMID- 17411014 TI - Sampling for metabolome analysis of microorganisms. AB - In the present work we investigated the most commonly applied methods used for sampling of microorganisms in the field of metabolomics in order to unravel potential sources of error previously ignored but of utmost importance for accurate metabolome analysis. To broaden the significance of our study, we investigated different Gram-negative and Gram-positive bacteria, i.e., Bacillus subtilis, Corynebacterium glutamicum, Escherichia coli, Gluconobacter oxydans, Pseudomonas putida, and Zymononas mobilis, and analyzed metabolites from different catabolic and anabolic intracellular pathways. Quenching of cells with cold methanol prior to cell separation and extraction led to drastic loss (>60%) of all metabolites tested due to unspecific leakage. Using fast filtration, Gram negative bacteria also revealed a significant loss (>80%) when inappropriate washing solutions with low ionic strength were applied. Adapting the ionic strength of the washing solution to that of the cultivation medium could almost completely avoid this problem. Gram-positive strains did not show significant leakage independent of the washing solution. Fast filtration with sampling times of several seconds prior to extraction appears to be a suitable approach for metabolites with relatively high intracellular level and low turnover such as amino acids or TCA cycle intermediates. Comparison of metabolite levels in the culture supernatant and the cell interior revealed that the common assumption of whole broth quenching protocols attributing the metabolites found exclusively to the intracellular pools may not be valid in many cases. In such cases a differential approach correcting for medium-contained metabolites is required. PMID- 17411015 TI - Utility of CE-MS data in protein identification. AB - A new method for displaying CE-MALDI-MS data for proteolytic digests is described. This data display mode yields distinct charge-based trends for plots of mass-to-charge (m/z) versus CE migration time. These trends arise owing to the in-solution charge state of the peptides, and this interpretation was confirmed by using empirical peptide electromigration models and peptide standards as charge-state markers. These charge-state specific trends exhibit analytical utility by providing additional chemical information about the peptides, which increases the confidence level of peptide identification and provides a rapid means for screening for posttranslationally modified peptides. PMID- 17411016 TI - Ultrahigh mass resolution and accurate mass measurements as a tool to characterize oligomers in secondary organic aerosols. AB - Organic aerosols are a major fraction, often more than 50%, of the total atmospheric aerosol mass. The chemical composition of the total organic aerosol mass is poorly understood, although hundreds of compounds have been identified in the literature. High molecular weight compounds have recently gained much attention because this class of compounds potentially represents a major fraction of the unexplained organic aerosol mass. Here we analyze secondary organic aerosols, generated in a smog chamber from alpha-pinene ozonolysis with ultra high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). About 450 compounds are detected in the mass range of m/z 200-700. The mass spectrum is clearly divided into a low molecular weight range (monomer) and a high molecular weight range, where dimers and trimers are distinguishable. Using the Kendrick mass analysis, the elemental composition of about 60% of all peaks could be determined throughout the whole mass range. Most compounds have high O:C ratios between 0.4 and 0.6. Small compounds (i.e., monomers) have a higher maximum O:C ratio than dimers and trimers, suggesting that condensation reactions with, for example, the loss of water are important in the oligomer formation process. A program developed in-house was used to determine exact mass differences between peaks in the monomer, dimer, and trimer mass range to identify potential monomer building blocks, which form the co-oligomers observed in the mass spectrum. A majority of the peaks measured in the low mass region of the spectrum (m/z < 300) is also found in the calculated results. For the first time the elemental composition of the majority of peaks over a wide mass range was determined using advanced data analysis methods for the analysis of ultra high-resolution MS data. Possible oligomer formation mechanisms in secondary organic aerosols were investigated. PMID- 17411017 TI - DFT energy surfaces for aminopurine homodimers and their conjugate acid ions. AB - Dimers of free nucleobases with their conjugate acid ions can be assigned to either of two categories: protonated dimers or proton-bound dimers. In the former, the extra proton attaches to a lone pair of a neutral dimer. In the latter, the extra proton is situated between two lone pairs and participates in a proton bridge. In general, proton-bound dimers are found to be more tightly held together than protonated dimers. While neutral adenine and its isomer 8 aminopurine (C(5)H(5)N(5)) are substantially more stable than their 7H tautomers, their conjugate acid ions and those of their respective 7H tautomers have nearly the same heats of formation. Correspondingly, the most stable (C(5)H(5)N(5))2H+ structures contain 7H tautomers as the neutral partner. Proton transit from one partner to the other within the most stable protonated dimer of 8-aminopurine has a low barrier (6 kJ mol(-1)). The potential energy curve for the NH stretch in that case is better fitted as a double minimum rather than as a harmonic potential. Purine-purine mismatches have been observed in nucleic acids, to which calculated (C(5)H(5)N(5))2H+ dimer geometries appear nearly isosteric. PMID- 17411018 TI - Heterogeneous reactions of gaseous HNO3 and NO2 on the clay minerals kaolinite and pyrophyllite. AB - Airborne clay mineral particles have long atmospheric lifetimes due to their relatively small size. To assess their impact on trace atmospheric gases, we investigated heterogeneous reactions on prototype clay minerals. Diffuse reflectance infrared spectroscopy identified surface-adsorbed products formed from the uptake of gaseous nitric acid and nitrogen dioxide on kaolinite and pyrophyllite. For kaolinite, a 1:1 phyllosilicate, HNO3 molecularly adsorbed onto the octahedral aluminum hydroxide and tetrahedral silicon oxide surfaces. Also detected on the aluminum hydroxide surface were irreversibly adsorbed monodentate, bidentate, bridged, and water-coordinated nitrate species as well as surface-adsorbed water. Similar adsorbed products formed during the uptake of NO2 on kaolinite at relative humidity (RH) of 0%, and the reaction was second order with respect to reactive surface sites and 1.5 +/- 0.1 for NO2. Reactive uptake coefficients, calculated using Brunauer, Emmett, and Teller surface areas, increased from (8.0 +/- 0.2) x 10(-8) to (2.3 +/- 0.4) x 10(-7) for NO2 concentrations ranging from 0.56 x 10(13) to 8.8 x 10(13) molecules cm(-3). UV visible spectroscopy detected gaseous HONO as a product for the reaction of NO2 on wet kaolinite. The uptake of HNO3 on pyrophyllite, a 2:1 phyllosilicate, resulted in stronger signal for nitric acid molecularly adsorbed on the silicon oxide surface compared to kaolinite. Monodentate, bridged, and water-coordinated nitrate species bound to aluminum sites also formed during this reaction indicating that reactive sites on edge facets are important for this system. The uptake of NO2 on pyrophyllite, gammaBET = (7 +/- 1) x 10(-9), was significantly lower than kaolinite because NO2 did not react with the dominant tetrahedral silicon oxide surface. These results highlight general trends regarding the reactivity of tetrahedral silicon oxide and octahedral aluminum hydroxide clay surfaces and indicate that the heterogeneous chemistry of clay aerosols varies with mineralogy and cannot be predicted by elemental analysis. PMID- 17411019 TI - Dehydrogenation of methanol by vanadium oxide and hydroxide cluster cations in the gas phase. AB - Bare vanadium oxide and hydroxide cluster cations, V(m)O(n)+ and V(m)O(n-1) (OH)+ (m = 1-4, n = 1-10), generated by electrospray ionization, were investigated with respect to their reactivity toward methanol using mass spectrometric techniques. Several reaction channels were observed, such as abstraction of a hydrogen atom, a methyl radical, or a hydroxymethyl radical, elimination of methane, and adduct formation. Moreover, dehydrogenation of methanol to generate formaldehyde was found to occur via four different pathways. Formaldehyde was released as a free molecule either upon transfer of two hydrogen atoms to the cluster or upon transfer of an oxygen atom from the cluster to the neutral alcohol concomitant with elimination of water. Further, formaldehyde was attached to V(m)O(n)+ upon loss of H2 or neutral water to produce the cation V(m)O(n)(OCH(2))+ or V(m)O(n-1) (OCH(2))+, respectively. A reactivity screening revealed that only high-valent vanadium oxide clusters are reactive with respect to H2 uptake, oxygen transfer, and elimination of H2O, whereas smaller and low-valent cluster cations are capable of dehydrogenating methanol via elimination of H2. For comparison, the reactivity of methanol with the corresponding hydroxide cluster ions, V(m)O(n-1) (OH)+, was studied also, for which dominant pathways lead to both condensation and association products, i.e., generation of the ions V(m)O(n-1) (OCH(3))+ and V(m)O(n-1) (OH)(CH(3)OH)+, respectively. PMID- 17411020 TI - Aggregation properties of heavy atom substituted squaraine dyes: evidence for the formation of J-type dimer aggregates in aprotic solvents. AB - The squaraine dye bis(2,4,6-trihydroxyphenyl)squaraine (SqH) was earlier reported to form J-type dimer aggregates in acetonitrile solutions at higher concentrations. Subsequent studies have suggested that concentration-dependent changes in the absorption spectrum of SqH in acetonitrile could be attributed to shifts in the acid-base equilibrium due to the presence of water as an impurity. In this work, we describe our studies on the effect of varying acid and dye concentration on the absorption spectra of the bromo and iodo substituted dyes, bis(3,5-dibromo-2,4,6-trihydroxyphenyl)squaraine (SqBr) and bis(3,5-diiodo-2,4,6 trihydroxyphenyl)squaraine (SqI). Analysis of the changes in the absorption spectra as a function of dye concentration and the nature of the solvent composition confirmed the formation of J-type dimer aggregates in aprotic solvents in this class of dyes. Further confirmation for the formation of the J type dimer aggregates could be obtained by comparing the differences in the triplet excited state properties of the neutral and aggregated forms of the dyes using time-resolved spectroscopy. PMID- 17411021 TI - Density-functional, density-functional tight-binding, and wave function calculations on biomolecular systems. AB - Recently, two computational approaches that supply a density-functional-based quantum-chemical method with an empirical term accounting for London dispersion were introduced and found use in the studies of biomolecular systems, namely, DFT D and SCC-DFTB-D. Here, we examine the performance and usability of these combined techniques for dealing with several tasks typically occurring in the research of biomolecules. The interaction energy of small biomolecular complexes agrees very well with the reference data yielded by correlated ab initio quantum chemical methods. In real-life studies aimed at interaction energy, structure, and infrared spectra, the mentioned methods provide results in good agreement with each other and with experiment (where available). The very favorable time demands of these approaches are discussed, and for each of them, a suitable area of use is proposed on the basis of the results of our analysis. PMID- 17411022 TI - Hydrogen bonds with pi and sigma electrons as the multicenter proton acceptors: high level ab initio calculations. AB - Pi-electrons of acetylene and sigma-electrons of molecular hydrogen were investigated as Lewis bases in different complexes. Hence high level ab initio calculations were performed up to the MP2/6-311++G(3df,3pd) level of approximation. It was found that species analyzed possess characteristics typical for H-bonded systems. The Bader theory was additionally applied; bond paths between proton and pi-electrons of acetylene or sigma-electrons of molecular hydrogen were detected with the corresponding bond critical points attributed to the proton-acceptor interactions. Numerous correlations between topological, geometrical and energetic parameters were also found. For example, the H...pi or H...sigma interaction is stronger for the shorter corresponding distance between the proton and the middle of C[triple bond]C or H-H bond. It is connected with the greater elongation of C[triple bond]C or H-H bonds and the greater transfer of electron charge from the Lewis base to the Lewis acid. PMID- 17411023 TI - Photoelectron spectrum of isolated ion-pairs in ionic liquid vapor. AB - The gas-phase valence binding energy spectrum of isolated ion-pairs of the commonly used [1-ethyl-3-methylimidazolium][bis(trifluoromethylsulfonyl)imide)] room-temperature ionic liquid is obtained by photoionization of a molecular beam of ionic liquid vapor by extreme ultraviolet light. The isolated ion-pair nature of the ionic liquid vapor is corroborated by single photon ionization mass spectroscopy, complemented by computed vaporization energetics of ion-pairs and clusters of ion-pairs. The valence binding energy spectrum of the isolated ion pairs is discussed in comparison with available liquid-phase data and theoretical density functional theory calculations. PMID- 17411024 TI - Novel 3-O-glycosyl-3-demethylthiocolchicines as ligands for glycine and gamma aminobutyric acid receptors. AB - New 3-O-glycosyl-3-demethylthiocolchicines containing natural and unnatural sugar moieties were prepared and tested on gamma-aminobutyric acid (GABA) and strychnine-sensitive glycine receptors present in rat brain and spinal cord. Two different synthetic approaches were used with the readily available 3-O demethylthiocolchicine (1b) and thiocolchicoside (2a). Glycosyl compounds 2a-g were obtained from 1b and 1-fluorosugars 4. 6'-Heterosubstituted glycosyl compounds 6-12 and the 6'-desoxy derivative 2h were prepared from 2a. PMID- 17411025 TI - Synthesis, biological testing, and binding mode prediction of 6,9-diarylpurin-8 ones as p38 MAP kinase inhibitors. AB - Based on the purine scaffold of ATP, derivatives of 6,9-diarylpurine-8-one were prepared and tested for their ability to inhibit p38 MAP kinase, a key enzyme in the cellular regulation of proinflammatory cytokines. The inhibitor design combines the purine system of the authentic cosubstrate ATP with various phenyl moieties to explore the selectivity for the two hydrophobic regions of the kinase's ATP-binding cleft. The present study indicates a new binding mode of our scaffold to p38 MAP kinase, which comprises the desired structural features of ATP and the N-phenyl-N-purin-6-yl ureas previously published by Wan et al. Combinations of Autodock and FlexX docking with different scoring functions were used to assess the postulated binding mode. The predictive power of different docking-scoring combinations was determined. The presented results may form a solid basis for further optimization cycles since our theoretical findings are consistent with our experimental binding data and supported by the literature. PMID- 17411026 TI - Novel styrylpyridines as probes for SPECT imaging of amyloid plaques. AB - We report a series of radioiodinated styrylpyridines as single photon emission computed tomography probes for imaging Abeta plaques in the brain of patients with Alzheimer's disease (AD). In vitro binding showed that all of the styrylpyridines displayed very good binding affinities in postmortem AD brain homogenates (Ki = 3.6 to 15.5 nM). No-carrier-added samples of 13a, 13b, 16a, 16b, and 16e (radioiodinated with 125I) were successfully prepared. The in vivo biodistribution in normal mice, at 2 min after injection, showed excellent initial brain penetrations (4.03, 6.22, 5.43, and 8.04% dose/g for [125I]13a, 13b, 16a, and 16b, respectively). Furthermore, in vitro autoradiography of AD brain sections showed that the high binding signal was specifically due to the presence of Abeta plaques. Taken together, these results strongly suggest that these styrylpyridines are useful for imaging Abeta plaques in the living human brain. PMID- 17411027 TI - Scaffold targeting drug-resistant colon cancers. AB - We have identified five derivatives of the natural product sansalvamide A that are potent against multiple drug-resistant colon cancer cell lines. These analogs share no structural homology to current colon cancer drugs, are cytotoxic at levels on par with existing drugs treating other cancers, and demonstrate selectivity for drug-resistant colon cancer cell lines over noncancerous cell lines. Thus, we have established sansalvamide A as a privileged structure for treating multiple drug-resistant colon cancers. PMID- 17411028 TI - Conformational sampling of bioactive molecules: a comparative study. AB - The necessity to generate conformations that sample the entire conformational space accessible to a given molecule is ubiquitous in the field of computer-aided drug design. Protein-ligand docking, 3D database searching, and 3D QSAR are three commonly used techniques that depend critically upon the quality and diversity of the generated conformers. Although there are a wide range of conformational search algorithms available, the extent to which they sample conformational space is often unclear. To address this question, we conducted a robust comparison of the search algorithms implemented in several widely used molecular modeling packages, including Catalyst, Macromodel, Omega, MOE, and Rubicon as well as our own method, stochastic proximity embedding (SPE). We found that SPE used in conjunction with conformational boosting, a heuristic for biasing conformational search toward more extended or compact geometries, along with Catalyst, are significantly more effective in sampling the full range of conformational space compared to the other methods, which show distinct preferences for either more extended or more compact geometries. PMID- 17411029 TI - Thermodynamics and kinetics of the nickel(II)-salicylhydroxamic acid system. Phenol rotation induced by metal ion binding. AB - The kinetics and the equilibria of Ni(II) binding to p-hydroxybenzohydroxamic acid (PHBHA) and salicylhydroxamic acid (SHA) have been investigated in an aqueous solution at 25 degrees C and I=0.2 M by the stopped-flow method. Two reaction paths involving metal binding to the neutral acid and to its anion have been observed. Concerning PHBHA, the rate constants of the forward and reverse steps are k1=(1.9+/-0.1)x10(3) M-1 s-1 and k-1=(1.1+/-0.1)x10(2) s-1 for the step involving the undissociated PHBHA and k2=(3.2+/-0.2)x10(4) M-1 s-1 and k-2=1.2+/ 0.2 s-1 for the step involving the anion. Concerning SHA, the analogous rate constants are k1=(2.6+/-0.1)x10(3) M-1 s-1, k-1=(1.3+/-0.1)x10(3) s-1, k2=(5.4+/ 0.2)x10(3) M-1 s-1, and k-2=6.3+/-0.5 s-1. These values indicate that metal binding to the anions of the two acids concurs with the Eigen-Wilkins mechanism and that the phenol oxygen is not involved in the chelation. Moreover, a slow effect was observed in the SHA-Ni(II) system, which has been put down to rotation of the benzene ring around the C-C bond. Quantum mechanical calculations at the B3LYP/lanL2DZ level reveal that the phenol group in the most stable form of the Ni(II) chelate is in trans position relative to the carbonyl oxygen, contrary to the free SHA structure, where the phenol and carbonyl oxygen atoms both have cis configuration. These results bear out the idea that the complex formation is coupled with phenol rotation around the C-C bond. PMID- 17411030 TI - Crystal structures of incommensurately modulated Ln(PO3)3 (Ln=Tb-Yb) and commensurate Gd(PO3)3 and Lu(PO3)3. AB - The crystal structure of the late lanthanoids' catena-polyphosphates Ln(PO3)3 (Ln=Tb-Yb) is incommensurately modulated (Dy(PO3)3: space group Cc(0beta0)0; Z=4; a=1417.4(4), b=670.96(14), c=1009.5(3) pm; beta=127.62(2) degrees, q=0.364b*; Rall=0.057, wRall=0.071; 293 K) and consists of infinite chains of corner-sharing PO4 tetrahedra. The cations are coordinated 6-fold in an almost octahedral arrangement over the whole modulation period. All atoms comprise a sinoidal positional modulation. The basic structure can be derived from a fcc packing which explains the pseudo-face-centering observed in the diffraction patterns. The crystal structure of Lu(PO3)3 is isotypic with C-type phosphates (Cc; Z=12; a=1397.2(1), b=2001.8(2), c=995.56(9) pm; beta=127.351(6) degrees; R1=0.042, wR2=0.097, 293 K), and Gd(PO3)3 crystallizes in a new structure type (I2/a; Z=16; a=2601.7(2), b=1351.1(1), c=1008.4(1) pm; beta=119.311(6) degrees; R1=0.039, wR2=0.092; 293 K). Both can be described in terms of superstructures of the basic structure unit cell of the incommensurate phases, and thus, a consistent structural description of many polyphosphates is provided. Tb(PO3)3 was obtained as single phase adopting a novel synthesis under reducing conditions. The absence of an inversion center in the incommensurate phases and Lu(PO3)3 was proved by a SHG experiment. The vibrational spectra are also discussed. PMID- 17411031 TI - Relating structural and thermodynamic effects of the Pb(II) lone pair: a new picolinate ligand designed to accommodate the Pb(II) lone pair leads to high stability and selectivity. AB - The crystal and molecular structure and the stability of lead and calcium complexes of two chelates containing picolinate chelating groups in different geometries have been investigated in order to relate the ligand affinity and selectivity for lead over calcium with the ability of the ligand to accommodate a stereochemically active lone pair. The crystal structures of the lead complexes of the diprotonated and monoprotonated tripodal ligand tpaa2- show that the three picolinate arms of the tripodal ligand coordinate the lead in an asymmetric way leaving a gap in the coordination sphere to accommodate the lead lone pair. As a consequence of this binding mode, one picolinate arm is very weakly bound and therefore can be expected to contribute very little to the complex stability. Conversely, the geometry of the dipodal ligand H2dpaea is designed to accommodate the lead lone pair; in the structure of the [Pb(dpaea)] complex the donor atoms of the ligand occupy only a quarter of the coordination sphere, reducing the sterical interaction between the lead lone pair with respect to the H3tpaa complexes. As a result, in the lead structures of H2dpaea all the ligand donor atoms are strongly bound to the metal ion leading to increased stability. The high value of the formation constant measured for the lead complex of the dipodal dpaea2- (log beta11(Pb)=12.1(3)) compared to the lower value found for the one of the tripodal tpaa3- (log beta11(Pb)=10.0(1)) provides direct evidence of the influence of the stereochemically active lead lone pair on complex stability. As a result, since the ligand geometry has little effect on the stability of the calcium complex, a remarkable increase in the Pb/Ca selectivity is observed for dpaea-(10(6.6)) compared to tpaa3- (10(1.5)), making the dipodal ligand a good candidate for application as extracting agent for the lead removal from contaminated water. PMID- 17411032 TI - Ligand topology variations and the importance of ligand field strength in non heme iron catalyzed oxidations of alkanes. AB - A series of iron(II)-bis(triflate) complexes [Fe(L)(OTf)2] containing linear tetradentate bis(quinolyl)-diamine and bis(quinolylmethyl)-diamine ligands with a range of ligand backbones has been prepared. The coordination geometries of these complexes have been investigated in the solid state by X-ray crystallography and in solution by 1H and 19F NMR spectroscopy. Because of the labile nature of high spin iron(II) complexes in solution, dynamic equilibria of complexes with different coordination geometries (cis-alpha, cis-beta, and trans) are observed with certain ligand systems. In these cases, the geometry observed in the solid state does not necessarily represent the only or even the major geometry present in solution. The ligand field strength in the various complexes has been investigated by variable-temperature (VT) magnetic moment measurements and by UV vis spectroscopy. The strongest ligand field is observed with the most rigid ligand that generates [Fe(L)(OTf)2] complexes with a cis-alpha coordination geometry, and the corresponding [Fe(L)(CH3CN)2]2+ complex displays spin crossover behavior. The catalytic properties of the complexes for the oxidation of cyclohexane have been investigated using hydrogen peroxide as the oxidant. An increased flexibility in the ligand results in a weaker ligand field, which increases the lability of the complexes. The activity and selectivity of the catalysts appear to be related to the strength of the ligand field and the stability of the catalyst. PMID- 17411033 TI - Formation of new halogenothiocarbonylsulfenyl halides, XC(S)SY, through photochemical matrix reactions starting from CS2 and a dihalogen molecule XY (XY=Cl2, Br2, or BrCl). AB - Isolation of a dihalogen molecule XY (XY=Cl2, Br2, or BrCl) with CS2 in a solid Ar matrix at about 15 K leads, by broad-band UV-vis photolysis (200500 microM). All beta-Lg derived peptides also exhibited radical-scavenging activity (oxygen radical absorbance capacity (ORAC) values ranged from 4.45 to 7.67 micromol Trolox equivalents/micromol of peptide). The presence and position of amino acids Trp, Tyr, and Met were proposed to be responsible for the antioxidant activity. The equimolar amino acid mixtures of all the peptides showed ORAC values lower than those of the corresponding peptides, indicating that the peptidic bond or the structural conformation had a positive influence on this activity. Finally, positive antioxidant effects of WYS, WYSL, and WYLA with ascorbic acid were observed, whereas WY and WYSLAM showed negative effects, both cases for different molar ratio mixtures. These results should be taken into account in the development of new food ingredients on the basis of peptides from beta-Lg. PMID- 17411067 TI - Binding of 2-nonanone and milk proteins in aqueous model systems. AB - Interactions of the model flavor compound 2-nonanone with individual milk proteins, whey protein isolate (WPI), and sodium caseinate in aqueous solutions were investigated. A method to quantify the free 2-nonanone was developed using headspace solid-phase microextraction followed by gas chromatography with flame ionization detection. Binding constants (K) and numbers of binding sites (n) for 2-nonanone on the individual proteins were calculated. The 2-nonanone binding capacities decreased in the order bovine serum albumin > beta-lactoglobulin > alpha-lactalbumin > alpha s1-casein > beta-casein, and the binding to WPI was stronger than the binding to sodium caseinate. All proteins appeared to have one binding site for 2-nonanone per molecule of protein at the flavor concentrations investigated, except for bovine serum albumin, which possessed two classes of binding sites. The binding mechanism is believed to involve predominantly hydrophobic interactions. PMID- 17411068 TI - Static-dynamic superheated liquid extraction of hydroxytyrosol and other biophenols from alperujo (a semisolid residue of the olive oil industry). AB - Hydroxytyrosol and other olive biophenols (OBPs) such as tyrosol, verbascoside, apigenin-7-glucoside, and alpha-taxifolin have been extracted from alperujo by using static-dynamic superheated liquids. Multivariate methodology has been used to carry out a detailed optimization of the extraction. Under the optimal working conditions no further extraction of the target analytes was achieved after 27 min (up to 2800 and 1500 mg/kg of hydroxytyrosol and tyrosol, respectively), so complete removal of them within this interval was assumed. The extract was injected into a chromatograph-photodiode array detector assembly for individual separation-quantification. The efficacy of ethanol/water mixtures to extract OBPs from alperujo has been demonstrated and compared with that of a conventional stirring-based method. These less toxic extractant mixtures are of interest with a view to future human uses of OBPs. PMID- 17411069 TI - Identification of antioxidants in essential oil of radix Angelicae sinensis using HPLC coupled with DAD-MS and ABTS-based assay. AB - Radix Angelicae sinensis (named danggui in Chinese), a commonly used traditional Chinese medicine, has multiple pharmacological activities. The essential oil of danggui is usually considered to be its main active fraction. However, to date, studies on the antioxidant potentials of danggui have focused on water-soluble compounds. In this paper, the antioxidant activity of the commercial essential oil of danggui was investigated by DPPH radical scavenging assay, ABTS radical scavenging assay, and beta-carotene bleaching test. Antioxidant constituents in the essential oil were identified using HPLC coupled with DAD-MS and ABTS-based assay. The results showed that the essential oil of danggui has concentration dependent antioxidant activity, which can be attributed to its component (coniferyl ferulate). This is the first report on the antioxidant activity of essential oil from danggui; its antioxidant potential was lower than those of positive controls, ascorbic acid and BHA. PMID- 17411070 TI - Antioxidant properties of Fusarium head blight-resistant and -susceptible soft red winter wheat grains grown in Virginia. AB - Fusarium head blight (FHB) has emerged as a major threat to wheat crops around the world, and it has been hypothesized that wheat antioxidants may play a role against Fusarium infections. The current study aimed to determine antioxidant properties of FHB-resistant wheat grains as compared to susceptible wheat. The wheat samples were collected from a single growing location (Warsaw, VA) and the same growing season. The results showed that both FHB-resistant and -susceptible wheat grains exerted strong radical scavenging activities against DPPH* radical [0.91-1.53 micromol of Trolox equivalents (TE)/g], peroxyl radical (15.5-24.5 micromol of TE/g), and hydroxyl radical (15.7-35.8 micromol of TE/g). Their total phenolic contents ranged from 888 to 1117 microg of gallic acid equivalents (GAE)/g. Five phenolic acids including ferulic, syringic, vanillic, caffeic, and p-coumaric acids were determined in soluble and insoluble fractions of wheat grains, altogether with a range of 219-389 microg/g. On average, the FHB resistant wheat group showed significantly higher average values in DPPH* and hydroxyl radicals scavenging activities (30 and 41% higher, respectively) than the FHB-susceptible wheat group. PMID- 17411071 TI - "One-pot" methylation in glycomics application: esterification of sialic acids and permanent charge construction. AB - A simple and rapid "one-pot" methylation method to esterify sialic acids and construct a permanent charge was developed for N-linked glycan analysis, which combined complete nonspecific proteolytic digestion and methylation. A mixture of Asn-glycans prepared from Pronase E digestion of the glycoprotein was passed through a cation-exchange column to convert carboxylic acids to the Na+ form before being methylated with methyl iodide. Derivatives could be easily purified with a hydrophilic affinity chromatography cartridge. Mass spectrometry analysis was performed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and MALDI-TOF/TOF. The mass spectrometric data indicated that carboxylic acids were methylated in addition to the formation of a quaternary ammonium in the amino group of asparagine residues. Three model glycoproteins, including ribonuclease B, ovalbumin, and transferrin, were employed to demonstrate the merits of this technique. Results showed that the stabilization of sialic acid was achieved in addition to the formation of a permanent charge. Compared to the analysis of underivatized N-glycans, detection sensitivity improved approximately 10-fold. The new technique was further evaluated with glycan profiling of serum transferrin and proved to be a sensitive method for the characterizing protein glycosylation. PMID- 17411072 TI - Comparison of the RNase H cleavage kinetics and blood serum stability of the north-conformationally constrained and 2'-alkoxy modified oligonucleotides. AB - The RNase H cleavage potential of the RNA strand basepaired with the complementary antisense oligonucleotides (AONs) containing North-East conformationally constrained 1',2'-methylene-bridged (azetidine-T and oxetane-T) nucleosides, North-constrained 2',4'-ethylene-bridged (aza-ENA-T) nucleoside, and 2'-alkoxy modified nucleosides (2'-O-Me-T and 2'-O-MOE-T modifications) have been evaluated and compared under identical conditions. When compared to the native AON, the aza-ENA-T modified AON/RNA hybrid duplexes showed an increase of melting temperature (DeltaTm = 2.5-4 degrees C per modification), depending on the positions of the modified residues. The azetidine-T modified AONs showed a drop of 4-5.5 degrees C per modification with respect to the native AON/RNA hybrid, whereas the isosequential oxetane-T modified counterpart, showed a drop of approximately 5-6 degrees C per modification. The 2'-O-Me-T and 2'-O-MOE-T modifications, on the other hand, showed an increased of Tm by 0.5 C per modification in their AON/RNA hybrids. All of the partially modified AON/RNA hybrid duplexes were found to be good substrates for the RNase H mediated cleavage. The Km and Vmax values obtained from the RNA concentration-dependent kinetics of RNase H promoted cleavage reaction for all AON/RNA duplexes with identical modification site were compared with those of the reference native AON/RNA hybrid duplex. The catalytic activities (Kcat) of RNase H were found to be greater (approximately 1.4-2.6-fold) for all modified AON/RNA hybrids compared to those for the native AON/RNA duplex. However, the RNase H binding affinity (1/Km) showed a decrease (approximately 1.7-8.3-fold) for all modified AON/RNA hybrids. This resulted in less effective (approximately 1.1-3.2-fold) enzyme activity (Kcat/Km) for all modified AON/RNA duplexes with respect to the native counterpart. A stretch of five to seven nucleotides in the RNA strand (from the site of modifications in the complementary modified AON strand) was found to be resistant to RNase H digestion (giving a footprint) in the modified AON/RNA duplex. Thus, (i) the AON modification with azetidine-T created a resistant region of five to six nucleotides, (ii) modification with 2'-O-Me-T created a resistant stretch of six nucleotides, (iii) modification with aza-ENA-T created a resistant region of five to seven nucleotide residues, whereas (iv) modification with 2'-O-MOE-T created a resistant stretch of seven nucleotide residues. This shows the variable effect of the microstructure perturbation in the modified AON/RNA heteroduplex depending upon the chemical nature as well as the site of modifications in the AON strand. On the other hand, the enhanced blood serum as well as the 3'-exonuclease stability (using snake venom phosphodiesterase, SVPDE) showed the effect of the tight conformational constraint in the AON with aza-ENA T modifications in that the 3'-exonuclease preferentially hydrolyzed the 3' phosphodiester bond one nucleotide away (n + 1) from the modification site (n) compared to all other modified AONs, which were 3'-exonuclease cleaved at the 3' phosphodiester of the modification site (n). The aza-ENA-T modification in the AONs made the 5'-residual oligonucleotides (including the n + 1 nucleotide) highly resistant in the blood serum (remaining after 48 h) compared to the native AON (fully degraded in 2 h). On the other hand, the 5'-residual oligonucleotides (including the n nucleotide) in azetidine-T, 2'-O-Me-T, and 2'-O-MOE-T modified AONs were more stable compared to that of the native counterpart but more easily degradable than that of aza-ENA-T containing AONs. PMID- 17411073 TI - Interplay of oxygen, vitamin E, and carotenoids in radical reactions following oxidation of Trp and Tyr residues in native HDL3 apolipoproteins. Comparison with LDL. A time-resolved spectroscopic analysis. AB - It has been recently shown that the inhibition of apolipoprotein A-I (apoAI) reverse cholesterol transport activity during oxidation of HDL by myeloperoxidase may involve myeloperoxidase electron transfer pathways other than those leading to tyrosine chlorination. To better understand how such mechanisms might be initiated, the role of semioxidized Tyr and Trp residues in loss of apoAI and apolipoprotein A-II (apoAII) integrity has been assessed using selective Trp and Tyr one-electron oxidation by *Br2(-) radical-anions in HDL3 as well as in unbound apoAI and apoAII. Behavior of these radicals in apolipoprotein B of LDL has also been assessed. Formation of semioxidized Tyr in HDL3 is followed by partial repair during several milliseconds via reaction with endogenous alpha tocopherol to form the alpha-tocopheroxyl radical. Subsequently, 2% of alpha tocopheroxyl radical is repaired by HDL3 carotenoids. With LDL, a faster repair of semioxidized Tyr by alpha-tocopherol is observed, but carotenoid repair of alpha-tocopheroxyl radical is not. Only a small fraction of HDL3 particles contains alpha-tocopherol and carotenoids, which explains limited repair of semioxidized Tyr by alpha-tocopherol. All LDL particles normally contain multiple alpha-tocopherol and carotenoid molecules, and the lack of repair of alpha tocopheroxyl radical by carotenoids probably results from hindered mobility of carotenoids in the lipid core. Western blots of gamma-irradiated HDL3 comparable to those reported for apoAI myeloperoxidase oxidation show that the incomplete repair of semioxidized Tyr and Trp induces apoAI and apoAII permanent damage including formation of a heterodimer of one apoAI with a monomeric apoAII at about 36 kDa. PMID- 17411074 TI - Probing the gamma2 calcium-binding site: studies with gammaD298,301A fibrinogen reveal changes in the gamma294-301 loop that alter the integrity of the "a" polymerization site. AB - To determine the significance of the gamma2 calcium-binding site in fibrin polymerization, we synthesized the fibrinogen variant, gammaD298,301A. We expected these two alanine substitutions to prevent calcium binding in the gamma2 site. We examined the influence of calcium on the polymerization of gammaD298,301A fibrinogen, evaluated its plasmin susceptibility, and solved 2.7 and 2.4 A crystal structures of the variant with the peptide ligands Gly-Pro-Arg Pro-amide (GPRP) and Gly-His-Arg-Pro-amide (GHRP), respectively. We found that thrombin-catalyzed polymerization of gammaD298,301A fibrinogen was modestly impaired, whereas batroxobin-catalyzed polymerization was significantly impaired relative to normal fibrinogen. Notably, the influence of calcium on polymerization was the same for the variant and for normal fibrinogen. Fibrinogen gammaD298,301A was more susceptible to plasmin proteolysis in the presence of GPRP. This finding suggests structural changes in the near-by "a" polymerization site. Comparisons of the structures revealed minor conformational changes in the gamma294-301 loop that are likely responsible for the weakened "a" site. When considered altogether, the data suggest that the gamma2 calcium-binding site does not significantly modulate polymerization. We cannot, however, rule out the possibility that the weakened "a" polymerization site masks an important role for the gamma2 calcium-binding site in normal polymerization. Somewhat unexpectedly, the structure data showed that GPRP bound to the "b" site and induced the same local conformational changes as GHRP to this site. This structure shows that "A:b" interactions can occur and suggests that these may participate in normal polymerization. PMID- 17411075 TI - Conformational and thermodynamic control of electron transfer in neuronal nitric oxide synthase. AB - Multiple solution-state techniques have been employed in investigating the nature and control of electron transfer in the context of the proposed "domain shuffle hypothesis" for intraprotein electron transfer inferred from the crystal structure of the nitric oxide synthase reductase domain. NADPH analogues and fragments have been used to map those regions of this substrate that are important in eliciting a conformational change, observed in both the fluorescence emission of the flavin cofactors of the enzyme and the EPR spectra of the FMN flavosemiquinone state. EPR and UV-visible potentiometric methods have demonstrated a substantial calmodulin-dependent perturbation in the midpoint reduction potentials of the redox couples of both flavin cofactors, in contrast to a previous report [Noble, M. A., et al. (1999) Biochemistry 38, 16413-16418]. These studies support a model in which FMN domain mobility, triggered by Ca2+ calmodulin binding and antagonized by substrate binding, facilitates electron transfer in nitric oxide synthase through conformational change and effects a major change in the midpoint reduction potentials of the flavin redox couples. These results are discussed in light of the recent crystal structure of the NADPH locked reductase domain. PMID- 17411076 TI - NikA binds heme: a new role for an Escherichia coli periplasmic nickel-binding protein. AB - NikA is a periplasmic binding protein involved in nickel uptake in Escherichia coli. NikA was identified as a heme-binding protein in the periplasm of anaerobically grown cells overexpressing CydDC, an ABC transporter that exports reductant to the periplasm. CydDC-overexpressing cells accumulate a heme biosynthesis-derived pigment, P-574. For further biochemical and spectroscopic analysis, unliganded NikA was overexpressed and purified. NikA was found to comigrate with both hemin and protoporphyrin IX during gel filtration. Furthermore, tryptophan fluorescence quenching titrations demonstrated that both hemin and protoporphyrin IX bind to NikA with similar affinity. The binding affinity of NikA for these pigments (Kd approximately 0.5 microM) was unaltered in the presence and absence of saturating concentrations of nickel, suggesting that these tetrapyrroles bind to NikA in a manner independent of nickel. To test the hypothesis that NikA is required for periplasmic heme protein assembly, the effects of a nikA mutation (nikA::Tn5, Km(R) insertion) on accumulation of P-574 by CydDC-overexpressing cells was assessed. This mutation significantly lowered P 574 levels, implying that NikA may be involved in P-574 production. Thus, in the reducing environment of the periplasm, NikA may serve as a heme chaperone as well as a periplasmic nickel-binding protein. The docking of heme onto NikA was modeled using the published crystal structure; many of the predicted complexes exhibit a heme-binding cleft remote from the nickel-binding site, which is consistent with the independent binding of nickel and heme. This work has implications for the incorporation of heme into b- and c-type cytochromes. PMID- 17411077 TI - Mass transport and electrode accessibility through periodic self-assembled nanoporous silica thin films. AB - Ordered nanoporous silica films have attracted great interest for their potential use to template nanowires for photovoltaics and thermoelectrics. However, it is crucial to develop films such that an electrode under the nanoporous film is accessible to solution species via facile mass transport through well-defined pores. Here, we quantitatively measure the electrode accessibility and the effective species diffusivity for nearly all the known nanoporous silica film structures formed by evaporation-induced self-assembly upon dip-coating or spin coating. Grazing-angle of incidence small-angle X-ray scattering was used to verify the nanoscale structure of the films and to ensure that all films were highly ordered and oriented. Electrochemical impedance spectroscopy (EIS) was then used to assess the transport properties. A model has been developed that separates the electrode/film kinetics and the film transport properties from the film/solution interface and bulk solution effects. Accounting for this, the accessible area of the nanoporous film coated FTO electrode (1-theta) is obtained from the high-frequency data, while the effective diffusivity of the ferrocene dimethanol (D(FDM)) redox couple is obtained from intermediate frequencies. It was found that the degree of order and orientation in the film, in addition to the symmetry/topology, is a dominant factor that determines these two key parameters. The EIS data show that the (211) oriented double gyroid, (110) oriented distorted body center cubic, and (211) distorted primitive cubic silica films have significant accessibility (larger than 26% of geometric area). However, the double-gyroid films showed the highest diffusivity by over an order of magnitude. Both the (10) oriented 2D hexagonal and (111) oriented rhombohedral films were found to be highly blocking with only small accessibility due to microporosity. The impedance data were also collected to study the stability of the nanoporous silica films in aqueous solutions as a function of pH. The distorted primitive silica film showed much faster degradation in pH 7 solution when compared to a blocking film such as the 2D hexagonal. However, silica films maintained their structure at pH 2 for at least 12 h. PMID- 17411078 TI - Effect of hydrophilic comonomer and surfactant type on the colloidal stability and size distribution of carboxyl- and amino-functionalized polystyrene particles prepared by miniemulsion polymerization. AB - Carboxyl and amino-functionalized polystyrene latex particles were synthesized by the miniemulsion copolymerization of styrene and acrylic acid or 2-aminoethyl methacrylate hydrochloride (AEMH). The reaction was started by using an oil soluble initiator, such as 2,2'-azobis(2-methylbutyronitrile) (V-59). The effect of the functional monomer content and type of surfactant (non-ionic versus ionic) on the particle size and particle size distribution was investigated by dynamic light scattering (DLS) and transmission electron microscopy (TEM). A bimodal particle size distribution was observed for functionalized latex particles prepared in the presence of the non-ionic surfactant (i.e., Lutensol AT-50) when 1 wt % of acrylic acid or 3 wt % of AEMH as a comonomer was employed. The copolymer particle nucleation was studied by using a highly hydrophobic fluorescent dye. From the obtained results, the formation of bimodal particle size distribution may be attributed to a budding-like effect, which takes place during the earlier stage of polymerization and is caused by the additional stabilizing energy originated from the ionic groups of a functional polymer. The reaction mechanism of particle formation in the presence of non-ionic and ionic surfactants has been proposed. The amount of the surface functional groups was determined from polyelectrolyte titration data. PMID- 17411079 TI - Adsorption and reaction of cyclohexene on a Ni(111) surface. AB - We studied the adsorption and reaction of cyclohexene (C6H10) on Ni(111) at different temperatures with high-resolution in-situ X-ray photoelectron spectroscopy (HR-XPS). For exposure at 125 K, we find intact cyclohexene with two distinct C 1s signals at 283.3 and 284.2 eV, due to the nonequivalent carbon atoms in the molecule. The energetic separation is significantly increased relative to the gas-phase value, due to the interaction with the substrate. Upon exposure at 210 K, complete dehydrogenation of cyclohexene to benzene (C6H6) and hydrogen is observed; coverage-dependent changes of the benzene adsorption site occur in a way similar to those for pure benzene layers, which indicates a phase separation in benzene and hydrogen islands. The thermal evolution of the adsorbed layers was studied by temperature-programmed (TP-) XPS and temperature-programmed desorption spectroscopy (TPD). Upon heating, the benzene + hydrogen layer formed at 210 K shows a coverage-dependent reorientation of the benzene molecules during partial desorption. The cyclohexene layer adsorbed at 125 K only shows partial conversion of cyclohexene to benzene and hydrogen upon heating to 185 or 210 K, with the remaining cyclohexene being stable up to approximately 300 K. We propose that upon heating these molecules are stabilized by coadsorbed benzene and hydrogen; furthermore, the mobility of benzene and hydrogen in this coadsorbed layer is reduced, so that no phase separation can occur. PMID- 17411080 TI - Inhibition of intrinsic proteolytic activities moderates preanalytical variability and instability of human plasma. AB - Human plasma and serum proteins are subject to intrinsic proteolytic degradation both during and after blood collection. By monitoring peptides, we investigated the stability of plasma and serum samples and the effects of anticoagulants and protease inhibitors on the plasma samples. Serum and plasma were subjected to time-course incubation, and the peptides (750-3200 Da) were extracted and analyzed with MALDI-TOF MS. Peptides of interest were further identified by MALDI TOF/TOF MS and ESI-MS/MS analyses. Our observations indicate that plasma peptides are significantly different from serum peptides. Intrinsic proteases cause these differences between plasma and serum samples, as well as the differences among three plasma samples using either EDTA, sodium citrate, or heparin as the anticoagulant, which accounts for partial inhibitory effects on plasma proteolytic activities. Proteases and peptidases, including both aminopeptidases and carboxypeptidases, also cause time-dependent, sequential generation and digestion of the peptides in serum and all three plasmas, specifically during early sample collection and processing. Protease inhibitors within an EDTA-plasma collection device inhibit both intrinsic plasma peptidases and proteases and moderate the time-dependent changes of the plasma peptides, including bradykinin, and complement C4- and C3- derived peptides. Our results suggest that mixing protease inhibitors immediately with blood during blood collection provides enhanced stabilization of the plasma proteome. PMID- 17411081 TI - Metabonomic investigations of aging and caloric restriction in a life-long dog study. AB - Long-term restriction of energy intake without malnutrition is a robust intervention that has been shown to prolong life and delay age-related morbidity. A 1H NMR-based metabonomic strategy was used to monitor urinary metabolic profiles throughout the lifetimes of control-fed and diet-restricted dogs. Urinary metabolic trajectories were constructed for each dog, and metabolic variation was found to be predominantly influenced by age. Urinary excretion of creatinine increased with age, reaching a maximum between ages 5 and 9 years and declining thereafter. Excretion of mixed glycoproteins was noted at earlier ages, which may be a reflection of growth patterns. In addition, consistent metabolic variation related to diet was also characterized, and energy-associated metabolites, such as creatine, 1-methylnicotinamide, lactate, acetate, and succinate, were depleted in urine from diet-restricted dogs. Both aging and diet restriction altered activities of the gut microbiotia, manifested by variation of aromatic metabolites and aliphatic amine compounds. This analysis allowed the metabolic response to two different physiological processes to be monitored throughout the lifetime of the canine population and may form part of a strategy to monitor and reduce the impact of age related diseases in the dog, as well as providing more general insights into extension of longevity in higher mammals. PMID- 17411082 TI - Photophysical properties of the newly synthesized triad based on [70]fullerene studies with laser flash photolysis. AB - N,N-Dimethylaniline-pyrazolinoC70-ferrocene has been prepared with the 1,3 dipolar cycloaddition reaction of a nitrile imine with C70. Although three regioisomers regarding the position of the pyrazolino group on the C70 were identified in the reaction products, molecular orbital calculations indicate that the stabilities and electronic properties of the three isomers are almost the same, which was confirmed by the sharp redox peaks. The photophysical properties of the triads have been investigated by measuring the time-resolved emission and transient absorption spectra showing that charge separation takes place efficiently via the photoexcited singlet state of the C70 moiety with accepting an electron from the donor moieties. It was found that the pyrazolino ring mediates a charge separation between the donor moieties and the photoexcited C70 moiety. PMID- 17411083 TI - Differences in solution behavior among four semiconductor-binding peptides. AB - Recent experiments have identified peptides that adhere to GaAs and Si surfaces. Here, we use all-atom Monte Carlo simulations with implicit solvent to investigate the behavior in aqueous solution of four such peptides, all with 12 residues. At room temperature, we find that all four peptides are largely unstructured, which is consistent with experimental data. At the same time, we find that one of the peptides is structurally different and more flexible, as compared to the others. This finding points at structural differences as a possible explanation for differences in adhesion properties among these peptides. By also analyzing designed mutants of two of the peptides, an experimental test of this hypothesis is proposed. PMID- 17411084 TI - Properties and structure of aromatic ester solvents. AB - This paper reports on an experimental and theoretical study of the aromatic ester solvents family. Several compounds were selected to analyze the different factors that influence their liquid-state properties and structures. The pressure-volume temperature behavior of these fluids was measured accurately over wide temperature and pressure ranges and correlated successfully with the empirical TRIDEN equation. From the measured data the relevant derived coefficients of isothermal compressibility, isobaric expansibility, and internal pressure were calculated. The statistical associating fluid theory (SAFT) and perturbed chain statistical associating fluid theory (PC-SAFT) molecularly based equations of state were used to predict the PVT behavior with model parameters obtained from the correlation of available saturation literature data; the results provided by PC-SAFT equations of state were clearly superior for all of the studied solvents. The fluid's molecular level structure was studied by quantum computations at the B3LYP/6-311++g** level and classical molecular dynamics simulations in the NPT ensemble with the OPLS-AA forcefield. Molecular parameters, such as torsional barriers or cluster energetics, were analyzed as a function of ester structures. The molecular dynamics study provides, on one hand, theoretical values of thermophysical properties, which are compared with the experimental ones, and, on the other hand, valuable molecular level structural information. On the basis of both macroscopic and microscopic studies complex fluid structures were inferred with important effects arising from the geometries of the studied molecules and from the existence of remarkable intermolecular forces of dominating dipolar nature. PMID- 17411085 TI - Stacking effects on local structure in RNA: changes in the structure of tandem GA pairs when flanking GC pairs are replaced by isoG-isoC pairs. AB - The Watson-Crick-like isoG-isoC (iGiC) pair, with the amino and carbonyl groups transposed relative to the Watson-Crick GC pair, provides an expanded alphabet for understanding interactions that shape nucleic acid structure. Here, thermodynamic stabilities of tandem GA pairs flanked by iGiC pairs are reported along with the NMR structures of the RNA self-complementary duplexes (GCiGGAiCGCA)2 and (GGiCGAiGCCA)2. A sheared GA pairing forms in (GCiGGAiCGCA)2, and an imino GA pairing forms in (GGiCGAiGCCA)2. The structures contrast with the formation of tandem imino and sheared GA pairs flanked by GC pairs in the RNA self-complementary duplexes (GCGGACGC)2 and (GGCGAGCC)2, respectively. In both iGiC duplexes, Watson-Crick-like hydrogen bonds are formed between iG and iC, and iGiC substitutions result in less favorable loop stability. The results provide benchmarks for testing computations of molecular interactions that shape RNA three-dimensional structure. PMID- 17411086 TI - Aqueous dispersions of C60 fullerene by use of amphiphilic block copolymers: preparation and nonlinear optical properties. AB - The dispersion of the otherwise insoluble C60 fullerene in water is discussed. Amphiphilic block copolymers, namely, polystyrene-b-polyethylene oxide (PS-PEO), were found to be able to disperse C60 in aqueous solutions, where the polymer forms micelles with a hydrophobic PS core. The preparation protocol of the final solutions was found to play a crucial role in the ability of the block copolymer to disperse the C60 molecules. The C60 containing aggregates were studied using optical spectroscopy, light scattering, and scanning electron microscopy. In addition, their optical limiting action and nonlinear optical properties under visible nanosecond laser excitation were studied and compared with that of C60 toluene solutions. PMID- 17411087 TI - Layer-by-layer assembled membranes of protonated 18-azacrown-6 and polyvinylsulfate and their application for highly efficient anion separation. AB - Ultrathin separation membranes were prepared upon alternating electrostatic adsorption of 1,4,7,10,13,16-hexaazacyclooctadecane (18-azacrown-6, hexacyclen, aza6) and polyvinylsulfate on porous polyacrylonitrile/polyethylene terephthalate (PAN/PET) substrates. The resulting composite membranes were highly permeable for electrolyte solutions, but the selectivity in ion transport was poor (alpha(Cl /SO42-) = 2). However, after a treatment of the membrane with 0.1 M aqueous copper(II) acetate solution, the rejection of divalent anions was strongly enhanced. Separation factors alpha(Cl-/SO42-) = 110 and alpha(Cl-/SO32-) = 1420 were found. Spectroscopic studies indicate a highly specific complex formation of aza6 and copper acetate in the membrane. Treatment of the membranes with cobalt(II) and nickel(II) acetate also led to an enhancement of the separation factors, but the effect was much smaller. Detailed studies of the influence of the pH of the dipping solutions and number of deposited bilayers are also reported. A model is presented describing the origin of the high transport selectivity. PMID- 17411088 TI - A monte carlo study of weak polyampholytes: stiffness and primary structure influences on titration curves and chain conformations. AB - The conformation and titration curves of weak polyampholytes are examined using Monte Carlo simulations with screened Coulomb potentials in the Grand Canonical ensemble. Two different types of monomers are considered. Depending on the solution pH, monomers A are weak acidic sites that can either be negatively charged or uncharged (as carboxylic groups), whereas monomers B are weak basic sites that can either be positively charged or uncharged (as amino groups). The influence of the chain stiffness, primary structure, and ionic concentration on the acid/base properties of the polyampholyte chains are systematically investigated. By adjusting the pH values, titration curves and then the fractions of positively and negatively ionized charged monomers are calculated. Stiffness influence is estimated by comparing two models of chain: a fully flexible and a rod-like polyampholyte. Different primary structures such as statistical (diblock, octablock, and alternating) and random polyampholytes are also considered. We demonstrate that the primary structure plays important roles in the acid/base properties as well as the charge distribution along the polymer backbone of a statistical rod-like polyampholyte. When flexible polyampholytes are considered, polyampholyte conformations promote the attractive electrostatic interactions between positively and negatively charged monomers, hence leading to more or less compact conformations and acid/base properties relatively different in comparison to the rod-like polyampholytes. Various conformations such as extended, globular, and pearl-necklace conformations are found in good agreement with the literature by adjusting the interaction parameter between monomers and monomer stoichiometry. PMID- 17411089 TI - Molecular solvation in water-methanol and water-sorbitol mixtures: the roles of preferential hydration, hydrophobicity, and the equation of state. AB - Molecular dynamics simulations of aqueous mixtures of methanol and sorbitol were performed over a wide range of binary composition, density (pressure), and temperature to study the equation of state and solvation of small apolar solutes. Experimentally, methanol is a canonical solubilizing agent for apolar solutes and a protein denaturant in mixed-aqueous solvents; sorbitol represents a canonical "salting-out" or protein-stabilizing cosolvent. The results reported here show increasing sorbitol concentration under isothermal, isobaric conditions results in monotonic increases in apolar solute excess chemical potential (mu2ex) over the range of experimentally relevant temperatures. For methanol at elevated temperatures, increasing cosolvent composition results in monotonically decreasing mu2ex. However, at lower temperatures mu2ex exhibits a maximum versus cosolvent concentration, as seen experimentally for Ar in ethanol-water solutions. Both density anomalies and hydrophobic effects--characterized by temperatures of density maxima and apolar solute solubility minima, respectively- are suppressed upon addition of either sorbitol or methanol at all temperatures and compositions simulated here. Thus, the contrasting effects of sorbitol and methanol on solute chemical potential cannot be explained by qualitative differences in their ability to enhance or suppress hydrophobic effects. Rather, we find mu2ex values across a broad range of temperatures and cosolvent composition can be quantitatively explained in terms of isobaric changes in solvent density--i.e., the equation of state--along with the corresponding packing fraction of the solvent. Analysis in terms of truncated preferential interaction parameters highlights that care must be taken in interpreting cosolvent effects on solvation in terms of local preferential hydration. PMID- 17411090 TI - In vitro studies of antifreeze glycoprotein (AFGP) and a C-linked AFGP analogue. AB - Antifreeze glycoproteins (AFGPs) are a subclass of biological antifreezes found in deep sea Teleost fish. These compounds have the ability to depress the freezing point of the organism such that it can survive the subzero temperatures encountered in its environment. This physical property is very attractive for the cryopreservation of cells, tissues, and organs. Recently, our laboratory has designed and synthesized a functional carbon-linked (C-linked) AFGP analogue (1) that demonstrates tremendous promise as a novel cryoprotectant. Herein we describe the in vitro effects and interactions of C-linked AFGP analogue 1 and native AFGP 8. Our studies reveal that AFGP 8 is cytotoxic to human embryonic liver and human embryonic kidney cells at concentrations higher than 2 and 0.63 mg/mL, respectively, whereas lower concentrations are not toxic. The mechanism of this cytotoxicity is consistent with apoptosis because caspase-3/7 levels are significantly elevated in cell cultures treated with AFGP 8. In contrast, C linked AFGP analogue 1 displayed no in vitro cytotoxicity even at high concentrations, and notably, caspase-3/7 activities were suppressed well below background levels in cell lines treated with 1. Although the results from these studies limit the human applications of native AFGP, they illustrate the benefits of developing functional C-linked AFGP analogues for various medical, commercial and industrial applications. PMID- 17411092 TI - Cells lacking DNA topoisomerase II beta are resistant to genistein. AB - Evidence suggests that DNA topoisomerases (topos) may be involved in the anticancer and carcinogenic properties attributed to flavonoids. Using the cell based assay TARDIS, the dietary flavonoids genistein (1) and luteolin (2) have been evaluated as topo I and topo II poisons and catalytic inhibitors in K562 leukemia cells. Both flavonoids induced topo II-DNA complexes, but they did not induce significant levels of topo I-DNA complexes. Genistein decreased the topo II-DNA complexes induced by the topo II poison etoposide, suggestive of a catalytic inhibition of topo II, and luteolin decreased the topo I-DNA complexes induced by the topo I poison camptothecin, indicative of a catalytic inhibition of topo I. Murine transgenic cells lacking topo II beta were resistant to genistein-induced cell growth inhibition (XTT assays) and cytotoxicity (clonogenic assay). High levels of topo II beta-DNA complexes were also observed in K562 cells exposed to genistein. These data suggest that topo II beta has an important function in genistein-induced cell growth inhibition and cell death. The possible role of topoisomerases in the putative anticancer and carcinogenic properties of genistein and luteolin is discussed. PMID- 17411091 TI - Rapid cross-linking of elastin-like polypeptides with (hydroxymethyl)phosphines in aqueous solution. AB - In situ gelation of injectable polypeptide-based materials is attractive for minimally invasive in vivo implantation of biomaterials and tissue engineering scaffolds. We demonstrate that chemically cross-linked elastin-like polypeptide (ELP) hydrogels can be rapidly formed in aqueous solution by reacting lysine containing ELPs with an organophosphorous cross-linker, beta [tris(hydroxymethyl)phosphino]propionic acid (THPP) under physiological conditions. The mechanical properties of the cross-linked ELP hydrogels were largely modulated by the molar concentration of lysine residues in the ELP and the pH at which the cross-linking reaction was carried out. Fibroblasts embedded in ELP hydrogels survived the cross-linking process and were viable after in vitro culture for 3 days. DNA quantification of ELP hydrogels with encapsulated fibroblasts indicated that there was no significant difference in DNA content between day 0 and day 3 when ELP hydrogels were formed with an equimolar ratio of THPP and lysine residues of the ELPs. These results suggest that THPP cross linking may be a biocompatible strategy for the in situ formation of cross-linked hydrogels. PMID- 17411093 TI - Tricalysiamides A-D, diterpenoid alkaloids from Tricalysia dubia. AB - Four rearranged ent-kaurane diterpenoid alkaloids, tricalysiamides A-D (1-4), having a cafestol-type carbon framework were isolated from the wood of Tricalysia dubia. Their absolute structures were determined on the basis of 2D NMR spectroscopy, X-ray crystallographic analysis, and chemical methods. PMID- 17411094 TI - Investigation of sulfur extrusion from a cyclic dialkoxy disulfide. AB - The relationship of cyclic dialkoxy disulfide 11, its thionosulfite isomer 12, and the related sulfoxylate 13 has been examined. Investigations demonstrate an interconversion between thionosulfite 12 and sulfoxylate 13. This sequential transformation brings evidence that a branched-bond sulfur structure is likely involved in sulfur extrusion. PMID- 17411095 TI - Nucleophilicities of primary and secondary amines in water. AB - The kinetics of the reactions of 26 primary and secondary amines with benzhydrylium ions in water were investigated photometrically. Because the parallel reactions of the benzhydrylium ions with hydroxide and water are much slower, the second-order rate constants for the reactions of amines with benzhydrylium ions could be determined reliably. Reactivities of anilines were also studied in acetonitrile solution. Plots of log k2,N for these reactions vs the electrophilicity parameters E of the benzhydrylium ions were linear, which allowed us to derive the nucleophilicity parameters N and s for amines as defined by the equation log k(20 degrees C)=s(E+N). Because the slope parameters for the different amines are closely similar; the relative nucleophilicities are almost independent of the electrophiles and can be expressed by the nucleophilicity parameters N. The correlation between nucleophilicity N and pKaH values is poor, and it is found that secondary alkyl amines and anilines are considerably more nucleophilic, while ammonia is much less nucleophilic than expected on the basis of their pKaH values. PMID- 17411096 TI - Zinc(II) perchlorate hexahydrate catalyzed opening of epoxide ring by amines: applications to synthesis of (RS)/(R)-propranolols and (RS)/(R)/(S)-naftopidils. AB - Commercially available zinc(II) perchlorate hexahydrate [Zn(ClO4)2.6H2O] was found to be a new and highly efficient catalyst for opening of epoxide rings by amines affording 2-amino alcohols in high yields under solvent-free conditions and with excellent chemo-, regio-, and stereoselectivities. For unsymmetrical epoxides, the regioselectivity was influenced by the electronic and steric factors associated with the epoxides and the amines. A complementarity in the regioselectivity was observed during the reaction of styrene oxide with aromatic and aliphatic amines: aromatic amines provided amino alcohols from nucleophilic attack at the benzylic carbon as major products whereas aliphatic amines resulted in formation of the amino alcohols through reaction at the terminal carbon atom of the epoxide ring as the major/sole products. Reaction of aniline with various glycidic ethers gave the amino alcohols by regioselective nucleophilic attack at the terminal carbon atom of the epoxide ring as the only/major product. Zinc(II) perchlorate hexahydrate was found to be the best catalyst compared to other metal perchlorates. The counteranion modulated the catalytic property of the various Zn(II) compounds that followed the order Zn(ClO4)2.(6)H2O>>Zn(BF4)2 approximately Zn(OTf)2>>ZnI2>ZnBr2>ZnCl2>Zn(OAc)2>Zn(CO3)2 in parallelism with the acidic strength of the corresponding protic acids (except for TfOH). The applicability of the methodology was demonstrated by the synthesis of cardiovascular drugs propranolol and naftopidil as racemates and optically active enantiomers. PMID- 17411097 TI - Facile synthesis of (+/-)-paeonilide. AB - (+/-)-Paeonilide, a novel monoterpenoid metabolite from the roots of Paeonia delavayi showing anti-platelet activating factor activity, is convergently synthesized in five steps with 59% overall yield. The application of benzoyl peroxide-promoted radical addition of unsaturated ester to aldehyde and subsequent topologically favored cyclization greatly simplified the synthesis. PMID- 17411098 TI - Parallel synthesis of an amide library based on the 6,8-dioxa-3 azabicyclo[3.2.1]octane scaffold by direct aminolysis of methyl esters. AB - An efficient synthesis of unsubstituted and substituted amides based on the 6,8 dioxa-3-azabicyclo[3.2.1]octane scaffold is described. The reaction, carried out at 60 degrees C in the absence of solvent, is characterized by its mildness and ease of workup. A library of amides, was synthesized by combination of methyl esters 1-6 with various amines. In addition, the microwave-assisted automated synthesis of the library was compared with the above conventional parallel synthesis. Microwave synthesis significantly decreased the reaction time from hours to minutes. PMID- 17411099 TI - A paramagnetic nanoprobe to detect tumor cell death using magnetic resonance imaging. AB - A 110 kDa (ca. 5 nm in diameter) bivalent paramagnetic nanoprobe for detecting cell death using magnetic resonance imaging (MRI) is described, in which two biotinylated C2A domains of the protein synaptotagmin-I were complexed with a single avidin molecule, which had been labeled with gadolinium chelates. This nanoprobe bound with high affinity and specificity to the phosphatidylserine exposed by dying cells and was demonstrated to allow MRI detection of apoptotic tumor cells in vitro. PMID- 17411100 TI - Overlapping resonances and Regge oscillations in the state-to-state integral cross sections of the F+H2 reaction. AB - A Regge pole analysis is employed to explain the oscillatory patterns observed in numerical simulations of integral cross section for the F+H(2)(v=0,j=0)- >HF(v(')=2,j(')=0)+H reaction in the translational collision energy range 25-50 meV. In this range the integral cross section for the transition, affected by two overlapping resonances, shows nearly sinusoidal oscillations below 38 meV and a more structured oscillatory pattern at larger energies. The two types of oscillations are related to the two Regge trajectories which (pseudo) cross near the energy where the resonances are aligned. Simple estimates are given for the periods of the oscillations. PMID- 17411101 TI - Efficient particle labeling in atomistic simulations. AB - The authors develop an efficient particle labeling procedure based on a linked cell algorithm which is shown to reduce the computing time for a molecular dynamics simulation by a factor of 3. They prove that the improvement of performance is due to the efficient fulfillment of both spatial and temporal locality principles, as implemented by the contiguity of labels corresponding to interacting atoms. Finally, they show that the present label reordering procedure can be used to devise an efficient parallel one-dimensional domain decomposition molecular dynamics scheme. PMID- 17411102 TI - Energy-consistent relativistic pseudopotentials and correlation consistent basis sets for the 4d elements Y-Pd. AB - Scalar-relativistic pseudopotentials and corresponding spin-orbit potentials of the energy-consistent variety have been adjusted for the simulation of the [Ar]3d(10) cores of the 4d transition metal elements Y-Pd. These potentials have been determined in a one-step procedure using numerical two-component calculations so as to reproduce atomic valence spectra from four-component all electron calculations. The latter have been performed at the multi-configuration Dirac-Hartree-Fock level, using the Dirac-Coulomb Hamiltonian and perturbatively including the Breit interaction. The derived pseudopotentials reproduce the all electron reference data with an average accuracy of 0.03 eV for configurational averages over nonrelativistic orbital configurations and 0.1 eV for individual relativistic states. Basis sets following a correlation consistent prescription have also been developed to accompany the new pseudopotentials. These range in size from cc-pVDZ-PP to cc-pV5Z-PP and also include sets for 4s4p correlation (cc pwCVDZ-PP through cc-pwCV5Z-PP), as well as those with extra diffuse functions (aug-cc-pVDZ-PP, etc.). In order to accurately assess the impact of the pseudopotential approximation, all-electron basis sets of triple-zeta quality have also been developed using the Douglas-Kroll-Hess Hamiltonian (cc-pVTZ-DK, cc pwCVTZ-DK, and aug-cc-pVTZ-DK). Benchmark calculations of atomic ionization potentials and 4d(m-2)5s(2)-->4d(m-1)5s(1) electronic excitation energies are reported at the coupled cluster level of theory with extrapolations to the complete basis set limit. PMID- 17411103 TI - Gyration-radius dynamics in structural transitions of atomic clusters. AB - This paper is concerned with the structural transition dynamics of the six-atom Morse cluster with zero total angular momentum, which serves as an illustrative example of the general reaction dynamics of isolated polyatomic molecules. It develops a methodology that highlights the interplay between the effects of the potential energy topography and those of the intrinsic geometry of the molecular internal space. The method focuses on the dynamics of three coarse variables, the molecular gyration radii. By using the framework of geometric mechanics and hyperspherical coordinates, the internal motions of a molecule are described in terms of these three gyration radii and hyperangular modes. The gyration radii serve as slow collective variables, while the remaining hyperangular modes serve as rapidly oscillating "bath" modes. Internal equations of motion reveal that the gyration radii are subject to two different kinds of forces: One is the ordinary force that originates from the potential energy function of the system, while the other is an internal centrifugal force. The latter originates from the dynamical coupling of the gyration radii with the hyperangular modes. The effects of these two forces often counteract each other: The potential force generally works to keep the internal mass distribution of the system compact and symmetric, while the internal centrifugal force works to inflate and elongate it. Averaged fields of these two forces are calculated numerically along a reaction path for the structural transition of the molecule in the three-dimensional space of gyration radii. By integrating the sum of these two force fields along the reaction path, an effective energy curve is deduced, which quantifies the gross work necessary for the system to change its mass distribution along the reaction path. This effective energy curve elucidates the energy-dependent switching of the structural preference between symmetric and asymmetric conformations. The present methodology should be of wide use for the systematic reduction of dimensionality as well as for the identification of kinematic barriers associated with the rearrangement of mass distribution in a variety of molecular reaction dynamics in vacuum. PMID- 17411104 TI - Car-Parrinello treatment for an approximate density-functional theory method. AB - The authors formulate a Car-Parrinello treatment for the density-functional-based tight-binding method with and without self-consistent charge corrections. This method avoids the numerical solution of the secular equations, the principal drawback for large systems if the linear combination of atomic orbital ansatz is used. The formalism is applicable to finite systems and for supercells using periodic boundary conditions within the Gamma-point approximation. They show that the methodology allows the application of modern computational techniques such as sparse matrix storage and massive parallelization in a straightforward way. All present bottlenecks concerning computer time and consumption of memory and memory bandwidth can be removed. They illustrate the performance of the method by direct comparison with Born-Oppenheimer molecular dynamics calculations. Water molecules, benzene, the C(60) fullerene, and liquid water have been selected as benchmark systems. PMID- 17411105 TI - Linear-scaling symmetric square-root decomposition of the overlap matrix. AB - We present a robust linear-scaling algorithm to compute the symmetric square-root or Lowdin decomposition of the atomic-orbital overlap matrix. The method is based on Newton-Schulz iterations with a new approach to starting matrices. Calculations on 12 chemically and structurally diverse molecules demonstrate the efficiency and reliability of the method. Furthermore, the calculations show that linear scaling is achieved. PMID- 17411106 TI - Nonequilibrium multiscale computational model. AB - A computational multiscale method is proposed to simulate coupled, nonequilibrium thermomechanical processes. This multiscale framework couples together thermomechanical equations at the coarse scale with nonequilibrium molecular dynamics at the fine scale. The novel concept of distributed coarse scale thermostats enables subsets of fine scale atoms to be attached to different coarse scale nodes which act as thermostats. The fine scale dynamics is driven by the coarse scale mean field. A coarse-grained Helmholtz free energy is used to derive macroscopic quantities. This new framework can reproduce the correct thermodynamics at the fine scale while providing an accurate coarse-grained result at the coarse scale. PMID- 17411107 TI - The electrostatic surface term: (I) periodic systems. AB - The authors propose a new approach to understand the electrostatic surface contributions to the interactions of large but finite periodic distributions of charges. They present a simple method to derive and interpret the surface contribution to any electrostatic field produced by a periodic distribution of charges. They discuss the physical and mathematical interpretations of this term. They present several examples and physical details associated with the calculation of the surface term. Finally, they provide a simple derivation of the surface contribution to the virial. This term does not disappear even if tinfoil boundary conditions are applied. PMID- 17411108 TI - Optimized theory for simple and molecular fluids. AB - An optimized closure approximation for both simple and molecular fluids is presented. A smooth interpolation between Perkus-Yevick and hypernetted chain closures is optimized by minimizing the free energy self-consistently with respect to the interpolation parameter(s). The molecular version is derived from a refinement of the method for simple fluids. In doing so, a method is proposed which appropriately couples an optimized closure with the variant of the diagrammatically proper integral equation recently introduced by this laboratory [K. M. Dyer et al., J. Chem. Phys. 123, 204512 (2005)]. The simplicity of the expressions involved in this proposed theory has allowed the authors to obtain an analytic expression for the approximate excess chemical potential. This is shown to be an efficient tool to estimate, from first principles, the numerical value of the interpolation parameters defining the aforementioned closure. As a preliminary test, representative models for simple fluids and homonuclear diatomic Lennard-Jones fluids were analyzed, obtaining site-site correlation functions in excellent agreement with simulation data. PMID- 17411109 TI - Exact stochastic simulation of coupled chemical reactions with delays. AB - Gillespie's exact stochastic simulation algorithm (SSA) [J. Phys. Chem. 81, 2350 (1977)] has been widely used to simulate the stochastic dynamics of chemically reacting systems. In this algorithm, it is assumed that all reactions occur instantly. While this is true in many cases, it is also possible that some chemical reactions, such as gene transcription and translation in living cells, take certain time to finish after they are initiated. Thus, the product of such reactions will emerge after certain delays. Apparently, Gillespie's SSA is not an exact algorithm for chemical reaction systems with delays. In this paper, the author develops an exact SSA for chemical reaction systems with delays, based upon the same fundamental premise of stochastic kinetics used by Gillespie in the development of his SSA. He then shows that an algorithm modified from Gillespie's SSA by Barrio et al. [PLOS Comput. Biol. 2, 1017 (2006)] is also an exact SSA for chemical reaction systems with delays, but it needs to generate more random variables than the author's algorithm. PMID- 17411110 TI - A simple density functional fractional occupation number procedure to determine the low energy transition region of spin-flip reactions. AB - A computationally simple three-step procedure to survey the energy landscape and to determine the molecular transition structure and activation energy at the intersection of two weakly coupled electronic potential energy surfaces of different symmetry is suggested. Only commercial software is needed to obtain the transition states of, for instance, spin-flip reactions. The computational expense is only two to three times larger than that of the standard determination of an adiabatic reaction path. First, the structures of the two electronic initial and final states along a chosen reaction coordinate are individually optimized. At the "projected crossing," the two states have the same energy at the same value of the reaction coordinate, but different state-optimized partial structures. Second, the unique optimized structure of a low energy crossing point between the two states is determined with the help of the density functional fractional occupation number approach. Finally, the respective energy of the two states at the crossing is estimated by a single point calculation. The prescription is successfully applied to some simple topical examples from organic and from inorganic chemistry, respectively, concerning the spin-flip reactions (3)H(3)CS(+)-->(1)H(2)CSH(+) and (7)MoCO(2)-->(5)MoCO(2)-->(3)OMoCO. PMID- 17411111 TI - Local integrals and their globally connected invariant structure in phase space giving rise to a promoting mode of chemical reaction. AB - We develop a method to extract local integrals, that is, integrals defined locally in the linear regime of an arbitrary point in phase space. The individual integral represents a vibrational mode. We also propose an index that quantifies the extent of connection between neighboring local integrals. Those pieces that are smoothly connected over a wide range represent a global structure of phase space. With a six-atomic Lennard-Jones cluster, we show that it is possible to identify which vibrational mode in the potential basin correlates smoothly to that in the area of transition state, which is nothing but a reactive mode. As an application of the method, we attempt to enhance the structural transition by exciting the reactive mode thus found. This method works successfully as shown in numerical calculations. PMID- 17411112 TI - Density scaling and relaxation of the Pauli principle. AB - The relaxation of the Pauli principle associated with density scaling is examined. Scaling the density has been investigated in the development of density functional computational methods with higher accuracy. Scaling the density by rho(r)(zeta)=rho(r)zeta reduces the number of electrons to M=Nzeta when zeta>1. The minimum kinetic energy of the scaled density, T(s)[rhozeta], can be scaled back to the N-electron system by multiplying the M-electron Kohn-Sham-type occupation numbers by zeta to produce T(zeta)[rho]. This relaxes the Pauli principle when the orbital occupation numbers are greater than 1 in the N electron system. The effects of antisymmetry on solutions to the Kohn-Sham equations are examined for Ne and the Be isoelectronic series. The changes in T(zeta)[rho] and the exchange energy E(xzeta)[rho] when zeta is varied show that these two quantities are inextricably linked. PMID- 17411113 TI - Polarization energy gradients in combined quantum mechanics, effective fragment potential, and polarizable continuum model calculations. AB - A method that combines quantum mechanics (QM), typically a solute, the effective fragment potential (EFP) discrete solvent model, and the polarizable continuum model is described. The EFP induced dipoles and polarizable continuum model (PCM) induced surface charges are determined in a self-consistent fashion. The gradients of these two energies with respect to molecular coordinate changes are derived and implemented. In general, the gradients can be formulated as simple electrostatic forces and torques among the QM nuclei, electrons, EFP static multipoles, induced dipoles, and PCM induced charges. Molecular geometry optimizations can be performed efficiently with these gradients. The formulas derived for EFPPCM can be generally applied to other combined molecular mechanics and continuum methods that employ induced dipoles and charges. PMID- 17411114 TI - Seams near seams: the Jahn-Teller effect in the 1E" state of N3+. AB - The (1)E(") electronic state of cyclic N(3) (+) arising from the singly excited electron configuration e('3)e(") is studied using multireference configuration interaction wave functions and a quadratic Jahn-Teller Hamiltonian determined from those calculations. It is shown that these two states have both a symmetry required seam of conical intersections at D(3h) geometries and three proximal symmetry equivalent C(2v) seams, located on a circle with radius rho(0) from the D(3h) intersection. rho(0), a function of Q(s), the breathing mode, is quite small but only attains a value of zero at Q(s) (crit)=1.252 A, resulting in a confluence or intersection node of the three C(2v) seams with the D(3h) seam. At this point only, g= parallelg(Q(s)) parallel, the norm of half the energy difference gradient, the linear Jahn-Teller term, vanishes and the intersection is of the Renner-Teller type. The close proximity of the previously unreported C(2v) seams to the D(3h) seam over the range of Q(s) considered is a consequence of the small values of g, compared to the quadratic Jahn-Teller term. The present analysis has important implications in the study of Jahn-Teller effects in ring systems and provides insight into a recent report that characterized this D(3h) seam as a Renner-Teller or glancing intersection. PMID- 17411115 TI - Polarizable atomic multipole solutes in a Poisson-Boltzmann continuum. AB - Modeling the change in the electrostatics of organic molecules upon moving from vacuum into solvent, due to polarization, has long been an interesting problem. In vacuum, experimental values for the dipole moments and polarizabilities of small, rigid molecules are known to high accuracy; however, it has generally been difficult to determine these quantities for a polar molecule in water. A theoretical approach introduced by Onsager [J. Am. Chem. Soc. 58, 1486 (1936)] used vacuum properties of small molecules, including polarizability, dipole moment, and size, to predict experimentally known permittivities of neat liquids via the Poisson equation. Since this important advance in understanding the condensed phase, a large number of computational methods have been developed to study solutes embedded in a continuum via numerical solutions to the Poisson Boltzmann equation. Only recently have the classical force fields used for studying biomolecules begun to include explicit polarization in their functional forms. Here the authors describe the theory underlying a newly developed polarizable multipole Poisson-Boltzmann (PMPB) continuum electrostatics model, which builds on the atomic multipole optimized energetics for biomolecular applications (AMOEBA) force field. As an application of the PMPB methodology, results are presented for several small folded proteins studied by molecular dynamics in explicit water as well as embedded in the PMPB continuum. The dipole moment of each protein increased on average by a factor of 1.27 in explicit AMOEBA water and 1.26 in continuum solvent. The essentially identical electrostatic response in both models suggests that PMPB electrostatics offers an efficient alternative to sampling explicit solvent molecules for a variety of interesting applications, including binding energies, conformational analysis, and pK(a) prediction. Introduction of 150 mM salt lowered the electrostatic solvation energy between 2 and 13 kcalmole, depending on the formal charge of the protein, but had only a small influence on dipole moments. PMID- 17411116 TI - Analytic derivatives for perturbatively corrected "double hybrid" density functionals: theory, implementation, and applications. AB - A recently proposed new family of density functionals [S. Grimme, J. Chem. Phys. 124, 34108 (2006)] adds a fraction of nonlocal correlation as a new ingredient to density functional theory (DFT). This fractional correlation energy is calculated at the level of second-order many-body perturbation theory (PT2) and replaces some of the semilocal DFT correlation of standard hybrid DFT methods. The new "double hybrid" functionals (termed, e.g., B2-PLYP) contain only two empirical parameters that have been adjusted in thermochemical calculations on parts of the G2/3 benchmark set. The methods have provided the lowest errors ever obtained by any DFT method for the full G3 set of molecules. In this work, the applicability of the new functionals is extended to the exploration of potential energy surfaces with analytic gradients. The theory of the analytic gradient largely follows the standard theory of PT2 gradients with some additional subtleties due to the presence of the exchange-correlation terms in the self-consistent field operator. An implementation is reported for closed-shell as well as spin unrestricted reference determinants. Furthermore, the implementation includes external point charge fields and also accommodates continuum solvation models at the level of the conductor like screening model. The density fitting resolution of the identity (RI) approximation can be applied to the evaluation of the PT2 part with large gains in computational efficiency. For systems with approximately 500-600 basis functions the evaluation of the double hybrid gradient is approximately four times more expensive than the calculation of the standard hybrid DFT gradient. Extensive test calculations are provided for main group elements and transition metal containing species. The results reveal that the B2 PLYP functional provides excellent molecular geometries that are superior compared to those from standard DFT and MP2. PMID- 17411117 TI - Doorway mechanism for dissociative electron attachment to fructose. AB - Recently, the three sugars ribose, deoxyribose, and fructose have been shown to undergo dissociative electron attachment at threshold, that is, to fragment upon capture of a zero-energy electron. Here the electron acceptor properties of three fructose isomers are investigated in view of a doorway mechanism. Two key ingredients for a doorway mechanism, a weakly bound state able to support a vibrational Feshbach resonance, and a valence anion more stable than neutral fructose are characterized. Moreover, possible structures for the observed fragment anion (fructose-H2O)- are suggested. PMID- 17411118 TI - State-to-state inelastic scattering of OH by HI: a comparison with OH-HCl and OH HBr. AB - Relative state-to-state cross sections and steric asymmetries have been measured for the scattering process: OH (X (2)Pi(32),v=0,J=32,M(J)=32,f)+HI ((1)Sigma,v=0,J<4)-->OH (X (2)Pi,v=0,Omega=12,J=12-52 and Omega=32,J=32 92,ef)+HI, at 690 cm(-1) collision energy. Comparison with the previously studied systems OH-HCl and OH-HBr reveals relevant features of the potential energy surfaces of these molecular systems. Some measured differences concerning the internal energy distribution after collision and the propensities for the impact with one or the other side of the OH molecule in scattering by HCl, HBr, and HI molecules are discussed. PMID- 17411119 TI - Fermi resonance in CO2: a combined electronic coupled-cluster and vibrational configuration-interaction prediction. AB - The authors present a first-principles prediction of the energies of the eight lowest-lying anharmonic vibrational states of CO(2), including the fundamental symmetric stretching mode and the first overtone of the fundamental bending mode, which undergo a strong coupling known as Fermi resonance. They employ coupled cluster singles, doubles, and (perturbative) triples [CCSD(T) and CCSDT] in conjunction with a range of Gaussian basis sets (up to cc-pV5Z, aug-cc-pVQZ, and aug-cc-pCVTZ) to calculate the potential energy surfaces (PESs) of the molecule, with the errors arising from the finite basis-set sizes eliminated by extrapolation. The resulting vibrational many-body problem is solved by the vibrational self-consistent-field and vibrational configuration-interaction (VCI) methods with the PESs represented by a fourth-order Taylor expansion or by numerical values on a Gauss-Hermite quadrature grid. With the VCI, the best theoretical estimates of the anharmonic energy levels agree excellently with experimental values within 3.5 cm(-1) (the mean absolute deviation). The theoretical (experimental) anharmonic frequencies of the Fermi doublet are 1288.9 (1285.4) and 1389.3 (1388.2) cm(-1). PMID- 17411120 TI - Spin-orbit effect in the energy pooling reaction O2(a 1Delta)+O2(a 1Delta)-->O2(b 1Sigma)+O2(X 3Sigma). AB - Five-dimensional nonadiabatic quantum dynamics studies have been carried out on two new potential energy surfaces of S(2)((1)A(')) and T(7)((3)A(")) states for the title oxygen molecules collision with coplanar configurations, along with the spin-orbit coupling between them. The ab initio calculations are based on complete active state second-order perturbation theory with the 6-31+G(d) basis set. The calculated spin-orbit induced transition probability as a function of collision energy is found to be very small for this energy pooling reaction. The rate constant obtained from a uniform J-shifting approach is compared with the existing theoretical and experimental data, and the spin-orbit effect is also discussed in this electronic energy-transfer process. PMID- 17411121 TI - Building transition probabilities for any condition using reduced cumulative energy transfer functions in H2O-H2O collisions. AB - The energy transfer process between highly vibrationally excited H(2)O in thermal equilibrium with a gas bath of H(2)O at different internal energies and temperatures has been studied by classical trajectory calculations. The results were analyzed using a cumulative probability distribution Q(DeltaE) of the amount of energy transferred, obtained by direct count of the number of trajectories that transfer an amount of energy equal to or greater than a certain value DeltaE. Scaling Q(DeltaE) in terms of the mean down and up energies transferred for each group of trajectories results in a unique distribution. This fact and the use of detailed balance constrains were used to propose a methodology that make it possible to build the whole P(E('),E) for any condition by knowing DeltaE and a series of parameters that depend only on the system under study. PMID- 17411122 TI - Intramolecular vibrational redistribution in Ne-Br2: the signature of intermediate resonances in the excitation spectrum. AB - Quantum-mechanical simulations of the Ne-Br(2)(B,v') excitation spectra produced after vibrational predissociation in the v'=20-35 range are reported. The aim is to investigate the signature in the excitation spectra of intermediate resonances lying in the lower vlambda(phot)>214 nm are reported. TKER spectra obtained at lambda(phot)>233 nm display broad, isotropic profiles that peak at low TKER ( approximately 1800 cm(-1)) and are largely insensitive to the choice of excitation wavelength. The bulk of these products is attributed to unintended multiphoton dissociation processes. TKER spectra recorded at lambda(phot)R(e)). The pronounced maxima in transition dipole moment (TDM) describing transitions into high electronic states (X (2)Pi-->3 (2)Pi, X (2)Pi-->3 (2)Sigma, 2 (2)Pi-->3 (2)Pi, 3 (2)Pi-->4 (2)Pi) occur at different interatomic separations. Such TDM features are promising for selection of excitation pathways and, consequently, for an optimal control of the dissociation products. PMID- 17411138 TI - Anomalous neutron Compton scattering cross sections in ammonium hexachlorometallates. AB - The authors have performed neutron Compton scattering measurements on ammonium hexachloropalladate (NH(4))(2)PdCl(6) and ammonium hexachlorotellurate (NH(4))(2)TeCl(6). Both substances belong to the family of ammonium metallates. The aim of the experiment was to investigate the possible role of electronic environment of a proton on the anomaly of the neutron scattering intensity. The quantity of interest that was subject to experimental test was the reduction factor of the neutron scattering intensities. In both samples, the reduction factor was found to be smaller than unity, thus indicating the anomalous neutron Compton scattering from protons. Interestingly, the anomaly decreases with decreasing scattering angle and disappears at the lowest scattering angle (longest scattering time). The dependence of the amount of the anomaly on the scattering angle (scattering time) is the same in both substances (within experimental error). Also, the measured widths of proton momentum distributions are equal in both metallates. This is consistent with the fact that the attosecond proton dynamics of ammonium cations is fairly well decoupled from the dynamics of the sublattice of the octahedral anions PdCl(6) (2-) and TeCl(6) (2 ), respectively. The hypothesis is put forward that proton-electron decoherence processes are responsible for the considered effect. Decoherence processes may have to do rather with the direct electronic environment of ammonium protons and not with the electronic structure of the metal-chlorine bond. PMID- 17411139 TI - First principles molecular dynamics of molten NaCl. AB - First principles Hellmann-Feynman molecular dynamics (HFMD) results for molten NaCl at a single state point are reported. The effect of induction forces on the structure and dynamics of the system is studied by comparison of the partial radial distribution functions and the velocity and force autocorrelation functions with those calculated from classical MD based on rigid-ion and shell model potentials. The first principles results reproduce the main structural features of the molten salt observed experimentally, whereas they are incorrectly described by both rigid-ion and shell-model potentials. Moreover, HFMD Green-Kubo self-diffusion coefficients are in closer agreement with experimental data than those predicted by classical MD. A comprehensive discussion of MD results for molten NaCl based on different ab initio parametrized polarizable interionic potentials is also given. PMID- 17411140 TI - Symmetry breaking of the fluid density profiles in closed nanoslits. AB - The density profiles in a fluid interacting with the two identical solid walls of a closed long slit were calculated for wide ranges of the number of fluid molecules in the slit and temperature by employing a nonlocal density functional theory. Using argon as the sample fluid and considering the walls composed of solid carbon dioxide, it is shown that the density profile corresponding to the stable state of the fluid considerably changes its shape with increasing average density rho(av) of the fluid inside the slit. Temperature dependent critical values rho(sb1) and rho(sb2) of rho(av) were identified, such that for rho(sb1)15 mT) and the IFF samples exhibit a clear hysteresis in the Raman spectra after releasing the magnetic field, which lasts for many hours at room temperature. The authors determined the characteristic times of the two competing processes, that of field-induced cluster formation and, at room temperature, that of thermal-activated dissociation, to range from 100 to 150 s. PMID- 17411147 TI - Liquid-vapor and liquid-liquid interfaces in Ising fluids: an integral equation approach. AB - The microscopic structure and thermodynamic properties of liquid-vapor and liquid liquid interfaces in Ising fluids are studied using an integral equation approach. The calculations are performed in the absence and presence of an external magnetic field by solving the corresponding set of Lovett-Mou-Buff Wertheim integrodifferential equations for the one-particle density distribution functions. The two-particle inhomogeneous direct correlation functions are consistently constructed by nonlinear interpolation between the bulk ones. The bulk correlation functions of the coexisting phases are obtained from the Ornstein-Zernike equations with a modified soft mean spherical approximation for the closure relation. As a result, the density and magnetization profiles at liquid-vapor and liquid-liquid interfaces as well as the surface tension and adsorption coefficients are evaluated in a wide temperature range including subcritical regions. The influence of an external magnetic field on the liquid vapor interfaces is also considered. PMID- 17411148 TI - Molecular dynamics methodology to investigate steady-state heterogeneous crystal growth. AB - In this paper a new molecular dynamics simulation methodology to investigate steady-state heterogeneous crystal growth from a supercooled liquid is presented. The method is tested on pure component systems such as Lennard-Jonesium and water/ice, as well as multicomponent systems such as methane hydrate crystals. The setup uses periodicity in all three directions and two interfaces; at one interface, crystallization occurs, while at the other, melting is enforced by locally heating the crystal only near that interface. Steady-state conditions are achieved when the crystal is melted at the same rate as the growth occurs. A self adaptive scheme that automatically modifies the rate of melting to match the rate of growth, crucial for establishing steady-state conditions, is described. In contrast with the recently developed method of Razul et al. [Mol. Phys. 103, 1929 (2005)], where the rates of growth (melting) were constant and the temperatures determined, the present approach fixes the supercooling temperature at the growing interface and identifies the corresponding steady-state crystal growth rate that corresponds to the thermodynamic force provided. The static properties of the interface (e.g., the interfacial widths) and the kinetics of the crystal growth are found to reproduce well previous findings. The importance of establishing steady-state conditions in such investigations is also briefly discussed. PMID- 17411149 TI - Strong correlations and Fickian water diffusion in narrow carbon nanotubes. AB - The authors have used atomistic molecular dynamics (MD) simulations to study the structure and dynamics of water molecules inside an open ended carbon nanotube placed in a bath of water molecules. The size of the nanotube allows only a single file of water molecules inside the nanotube. The water molecules inside the nanotube show solidlike ordering at room temperature, which they quantify by calculating the pair correlation function. It is shown that even for the longest observation times, the mode of diffusion of the water molecules inside the nanotube is Fickian and not subdiffusive. They also propose a one-dimensional random walk model for the diffusion of the water molecules inside the nanotube. They find good agreement between the mean-square displacements calculated from the random walk model and from MD simulations, thereby confirming that the water molecules undergo normal mode diffusion inside the nanotube. They attribute this behavior to strong positional correlations that cause all the water molecules inside the nanotube to move collectively as a single object. The average residence time of the water molecules inside the nanotube is shown to scale quadratically with the nanotube length. PMID- 17411150 TI - Bonding-site dependence of surface resistivity: CO on epitaxial Cu(100) films. AB - The authors have measured the contribution of CO adsorbed on different bonding sites to the resistivity of epitaxial Cu(100) films. Through an analysis of the correlation between surface resistivity and temperature-programed desorption measurements, CO on terraces is found to affect the resistance much more strongly than CO on defect sites. Two possible explanations are considered: a reduced scattering cross section of defect CO owing to the different local electronic structure at defect sites or strong scattering by the bare defect that is not significantly affected by the adsorption of CO. PMID- 17411151 TI - Ab initio study of the effects of orientation and corrugation for H2 adsorbed on polycyclic aromatic hydrocarbons. AB - This paper reports state-of-the-art ab initio calculations at the second order of Moller-Plesset perturbation theory of molecular hydrogen interaction with polycyclic aromatic hydrocarbons (PAHs) up to coronene (C(24)H(12)). The effects of both H(2) orientation with respect to the PAH plane and of PAH corrugation have been carefully investigated. It was found that the energetic preference for the perpendicular H(2) orientation over the parallel one decreases rapidly as the PAH grows. The detailed study of coronene-H(2) potential energy surface has made it possible to estimate from first principles the graphite-H(2) binding energy. This ab initio estimate is shown to be in fair agreement with experiment. As revealed by the example of benzene-H(2) dimer, the electrostatic energy component plays an important role in PAH-H(2) interactions, that stems largely from the charge penetration effect and, therefore, cannot be reproduced by the simple quadrupole-quadrupole interaction model. PMID- 17411152 TI - Theoretical and computational modeling of self-oscillating polymer gels. AB - The authors model wave propagation in swollen, chemoresponsive polymer gels that are undergoing the oscillatory Belousov-Zhabotinsky (BZ) reaction. To carry out this study, they first modify the Oregonator model for BZ reactions in simple solutions to include the effect of the polymer on the reaction kinetics. They then describe the gel dynamics through the framework of the two-fluid model. The polymer-solvent interactions that are introduced through the BZ reaction are captured through a coupling term, which is added to the Flory-Huggins model for polymer-solvent mixtures. The resulting theoretical model is then used to develop the gel lattice spring model (gLSM), which is a computationally efficient approach for simulating large-scale, two-dimensional (2D) deformations and chemical reactions within a swollen polymer network. The 2D calculations allow the authors to probe not only volume changes but also changes in the sample's shape. Using the gLSM, they determine the pattern formation and shape changes in 2D rectangular BZ gels that are anchored to a solid wall. They demonstrate that the dynamic patterns depend on whether the gel is expanded or contracted near the wall, and on the sample's dimensions. Finally, they isolate a scenario where the detachment of the gel from the wall leads to macroscopic motion of the entire sample. PMID- 17411153 TI - Molecular dynamics study of the stability of methane structure H clathrate hydrates. AB - Molecular dynamics simulations are used to study the stability of structure H (sH) methane clathrate hydrates in a 3 x 3 x 3 sH unit cell replica. Simulations are performed at experimental conditions of 300 K and 2 GPa for three methane intermolecular potentials. The five small cages of the sH unit cell are assigned methane guest occupancies of one and large cage guest occupancies of one to five are considered. Radial distribution functions, unit cell volumes, and configurational energies are studied as a function of large cage CH(4) occupancy. Free energy calculations are carried out to determine the stability of clathrates for large cage occupancies. Large cage occupancy of five is the most stable configuration for a Lennard-Jones united-atom potential and the Tse-Klein McDonald potential parametrized for condensed methane phases and two for the most stable configuation for the Murad and Gubbins potential. PMID- 17411154 TI - Electronic structure of copper phthalocyanine: an experimental and theoretical study of occupied and unoccupied levels. AB - An experimental and theoretical study of the electronic structure of copper phthalocyanine (CuPc) molecule is presented. We performed x-ray photoemission spectroscopy (XPS) and photoabsorption [x-ray absorption near-edge structure (XANES)] gas phase experiments and we compared the results with self-consistent field, density functional theory (DFT), and static-exchange theoretical calculations. In addition, ultraviolet photoelectron spectra (UPS) allowed disentangling several outer molecular orbitals. A detailed study of the two highest occupied orbitals (having a(1u) and b(1g) symmetries) is presented: the high energy resolution available for UPS measurements allowed resolving an extra feature assigned to vibrational stretching in the pyrrole rings. This observation, together with the computed DFT electron density distributions of the outer valence orbitals, suggests that the a(1u) orbital (the highest occupied molecular orbital) is mainly localized on the carbon atoms of pyrrole rings and it is doubly occupied, while the b(1g) orbital, singly occupied, is mainly localized on the Cu atom. Ab initio calculations of XPS and XANES spectra at carbon K edge of CuPc are also presented. The comparison between experiment and theory revealed that, in spite of being formally not equivalent, carbon atoms of the benzene rings experience a similar electronic environment. Carbon K-edge absorption spectra were interpreted in terms of different contributions coming from chemically shifted C 1s orbitals of the nonequivalent carbon atoms on the inner ring of the molecule formed by the sequence of CN bonds and on the benzene rings, respectively, and also in terms of different electronic distributions of the excited lowest unoccupied molecular orbital (LUMO) and LUMO+1. In particular, the degenerate LUMO appears to be mostly localized on the inner pyrrole ring. PMID- 17411155 TI - Dynamic behaviors on zadaxin getting into carbon nanotubes. AB - The dynamic behaviors of drug zadaxin getting into carbon nanotubes (CNTs) in different water surroundings were investigated by molecular dynamics simulation. It was found that the diameter (1.9 nm) of (14, 14) CNT is the critical size for inserting zadaxin into CNT at the present conditions. In addition, the length of CNTs is another factor for inserting. A certain length is needed. It implies that interactions of zadaxin with both the CNT and the water molecules are competitive in the insertion process. The CNT-zadaxin attractive interaction is found to be the main driving force with the lower density of water molecules in the surroundings, while the zadaxin-water interaction becomes dominant with the higher density. The study of the authors suggests that biomolecules-CNT systems can be further exploited for the potential applications to drugs, vaccines, and gene delivery. PMID- 17411156 TI - Self-assembling of ABC linear triblock copolymers in nanocylindrical tubes. AB - By employing Monte Carlo simulations for various tube diameters and preferences of the tube surface for the A, B, and C segments, the morphologies of A(5)B(5)C(5), A(5)B(10)C(5), and A(5)B(5)C(10) triblock copolymer melts confined in nanocylindrical tubes were examined. The interaction parameters between different segments were considered constant epsilon(AB)=epsilon(AC)=epsilon(BC)=0.3k(B)T, the tube diameter was changed from d=9xlattice parameter to d=33xlattice parameter, and the preferences of the tube surface for the segments A, B, and C (-epsilon(AS),-epsilon(BS), and epsilon(CS)) were varied between 0.05k(B)T and k(B)T. ABCCBA alternately stacked disks were generated in most tubes when the preference of the tube surface for any of the segments was weak, and the morphologies tended to transform into curved lamellae in tubes with large diameters when the preference for one of the segments was high. Numerous novel morphologies, such as ABC double helixes, AB single helix+C double helixes, AB double helixes+C quadruple helixes, plate morphologies with fins, dendrites, etc., which were located in the phase diagram between the stacked disks and the curved lamellar structures, were identified. Additionally, the orientation parameters indicating the alignments of the polymer chains were calculated and correlated with the morphologies. PMID- 17411157 TI - Molecular dynamics simulation of diffusion and sorption of water in conducting polyaniline. AB - Energy minimization and molecular dynamics simulations are used to develop, for the first time, atomistic models of HCl- and HBr-doped conducting polyanilines, in order to study diffusion and adsorption of water vapor in the polymers. Various morphological properties of the polymers are computed, including their pair correlation functions that are found to be in good agreement with the experimental data, and their accessible free volumes. Also computed are the sorption isotherms and effective self-diffusivity of water vapor in the polymers. The computed sorption isotherms are in quantitative agreement with the experimental data, while the diffusivities are within an order of magnitude of the data. The reasons for the differences between the computed and measured diffusivities are discussed. PMID- 17411158 TI - Polymer monolayers with a small viscoelastic linear regime: equilibrium and rheology of poly(octadecyl acrylate) and poly(vinyl stearate). AB - The equilibrium properties of monolayers of two polymers: poly(octadecyl acrylate) and poly(vinyl stearate) on water have been measured. The surface pressure (Pi) versus surface concentration (Gamma) curves indicate that the water air interface is a poor solvent for both polymers. The thermal expansivity shows a sharp change near room temperature. This behavior is typical of a glass transition; this is the first time that such a plot is observed for Langmuir films. The Pi vs Gamma curves measured by the continuous compression method show strong anisotropy effects. They also show that the monolayer is brought into nonequilibrium states depending on the compression rate. Within the linear regime, the relaxation experiments were bimodal. The longest relaxation time strongly increases as T is decreased, which might be compatible with the high increase of viscosity in the glass transition. The oscillatory barrier experiments showed that the maximum strain of the linear regime is smaller than 3% for both monolayers. The Fourier-transform analysis of the oscillatory experiments beyond the linear regime points out the contribution of different harmonics in the response function. Oscillations in the nonlinear regime show hysteresis cycles. The results obtained indicate that some of the previously published data for these polymer monolayers correspond to nonequilibrium states. PMID- 17411159 TI - Pressure driven flow of polymer solutions in nanoscale slit pores. AB - Polymer solutions subject to pressure driven flow and in nanoscale slit pores are systematically investigated using the dissipative particle dynamics approach. The authors investigated the effect of molecular weight, polymer concentration, and flow rate on the profiles across the channel of the fluid and polymer velocities, polymer density, and the three components of the polymers radius of gyration. They found that the mean streaming fluid velocity decreases as the polymer molecular weight and/or polymer concentration is increased, and that the deviation of the velocity profile from the parabolic profile is accentuated with increase in polymer molecular weight or concentration. They also found that the distribution of polymers conformation is highly anisotropic and nonuniform across the channel. The polymer density profile is also found to be nonuniform, exhibiting a local minimum in the center plane followed by two symmetric peaks. They found a migration of the polymer chains either from or toward the walls. For relatively long chains, as compared to the thickness of the slit, a migration toward the walls is observed. However, for relatively short chains, a migration away from the walls is observed. PMID- 17411160 TI - Simulation of nonlinear shear rheology of dilute salt-free polyelectrolyte solutions. AB - Brownian dynamics simulations are used to conduct a systematic analysis of the nonlinear shear rheology of dilute polyelectrolyte solutions, exploring its relationship to shear rate, Bjerrum length, and concentration. A simple coarse grained bead-spring chain model that incorporates explicit counterions is used. It is found that the polyelectrolyte chains exhibit a shear thinning behavior at high shear rate (as characterized by bead Peclet number Pe) that is independent of the electrostatic strength due to the stripping of ions from close proximity to the chain caused by the flow. In contrast, at low values of Pe, the viscosity increases monotonically with increasing Bjerrum length over the range studied here, in contrast to the nonmonotonic trend displayed by the chain size. Furthermore, at fixed Bjerrum length, the reduced viscosity increases monotonically with concentration. The mechanism underlying these observations is essentially the primary electroviscous effect; the ion cloud surrounding a polyelectrolyte chain deforms in flow, causing a significant increase in viscosity as concentration increases. Finally, the authors have also considered the role of hydrodynamic interactions in these simulations, finding that for low concentration studies in shear flow, these do not qualitatively affect the results. PMID- 17411161 TI - Rheology of concentrated carbon nanotube suspensions. AB - The rheological properties of non-Brownian carbon nanotube suspensions are measured over a range of nanotube volume fractions spanning the transition from semidilute to concentrated. The polymer-stabilized nanotubes are "sticky" and form a quiescent elastic network with a well-defined shear modulus and yield stress that both depend strongly on nanotube volume fraction with different but related critical exponents. We compare controlled-strain-rate and controlled stress measurements of yielding in shear flow, and we study the effect of slow periodic stress reversal on yielding and the arrest of flow. Our measurements support a universal scaling of both the linear viscoelastic and steady-shear viscometric response. The former allows us to extract the elastic shear modulus of semidilute nanotube networks for values that are near or below the resolution limit of the rheometers used, while the latter provides a similar extrapolation of the yield stress. A simple scaling argument is used to model the dependence of yield stress and elastic modulus on concentration. PMID- 17411162 TI - Energetics of the dipole flip-flop motion in a ferroelectric polymer chain. AB - The authors report on a study of dipole flip-flop "local" transition in ferroelectric polyvinylidene fluoride [P(VDF)] chains, using total energy calculation based on the density functional theory. The calculated results indicate that a simple flipping of a single electric dipole moment is energetically allowed. Furthermore, such a flipping involves no change either in bond length, bond angle, or the orientation of the chain. The calculations also show that on a thin film of ordered chains, strong dipole interactions existing in P(VDF) could cause modulation of the dipole orientation thus forming superlattices on P(VDF) films. These results are in good agreement with recent scanning tunnel microscope experimental measurements. Furthermore, our calculations show that partial flipping may also exist and extend over a length of several monomers during the flip-flop transition. PMID- 17411163 TI - Effective attraction interactions between like-charge macroions bound to binary fluid lipid membranes. AB - Using integral equation theory of liquids to a binary mixed fluid lipid membrane, the authors study the membrane-mediated interactions between binding macroions and the redistribution of neutral and charged lipids due to the macroions. The authors find that when the concentration of binding macroions is infinitely dilute, the main contribution to the attractive potential between macroions is the line tension between neutral and charged lipids of the membrane. As the relative concentration of charged lipids is increased, the authors observe a repulsive-attractive-repulsive potential transition due to the competition between the line tension of mixed lipids and screened electrostatic macroion macroion interactions. For the finite concentration of macroions, the main feature of the attraction is similar to the infinite-diluted case. However, the corresponding line tension of binary lipids under single macroion is lowered with the formation of multicomplexes by the charged lipids and the macroions, and the maximum of attractive potential will shift toward the higher values of charged lipid concentration. PMID- 17411164 TI - Application of the accelerated molecular dynamics simulations to the folding of a small protein. AB - In this paper, we further explore the applicability of the accelerated molecular dynamics simulation method using a bias potential. The method is applied to both simple model systems and real multidimensional systems. The method is also compared to replica exchange simulations in folding a small protein, Trp cage, using an all atom potential for the protein and an implicit model for the solvent. We show that the bias potential method allows quick searches of folding pathways. We also show that the choice of the bias potential has significant influence on the efficiency of the bias potential method. PMID- 17411165 TI - Inhomogeneity effects on the structure and dynamics of water at the surface of a membrane: a computer simulation study. AB - The authors report the structural and dynamical properties of water interacting with the surface of a lipid bilayer. Three regions have been identified, which show different dynamical regimes of water: a region of strong water-solute interaction, a transition region, and the bulk water region. The dynamics of the strong-interacting water is dominated by caging effects, as shown by the analysis of the self-intermediate scattering function, and by the disrupture of water's hydrogen bond network, while the smooth transition to bulk water is traced back to the roughness of the bilayer surface. PMID- 17411166 TI - Complex dynamics in a cross-catalytic self-replication mechanism. AB - The authors consider a minimal cross-catalytic self-replicating system of only two cross-catalytic templates that mimics the R3C ligase ribozyme system of Dong Eu and Joyce [Chem. Biol. 11, 1505 (2004)]. This system displays considerably more complex dynamics than its self-replicating counterpart. In particular, the authors discuss the Poincare-Andronov-Hopf bifurcation, canard transitions, excitability, and hysteresis that yield birhythmicity between simple and complex oscillations. PMID- 17411167 TI - Restricted open-shell Kohn-Sham theory: simulation of the pyrrole photodissociation. AB - The authors study the photodissociation reactions of pyrrole and N-methylpyrrole using first-principles molecular dynamics. The first excited state is described with restricted open-shell Kohn-Sham theory. They find a small barrier in the excited state potential energy surface. The possibility of energy redistribution near the Franck-Condon region leads to two different reaction channels in on-the fly simulations on a single diabatic potential energy surface. The results are discussed in comparison with previous ab initio calculations and with experiments. PMID- 17411168 TI - Electric quadrupole model on the formation of molecular chirality dependent domain shapes of lipid monolayers at the air-water interface. AB - Shapes and orientational deformation of a lipid monolayer domain have been analyzed taking into account the surface pressure, line tension, and electrostatic energy due to the spontaneous polarization and electric quadrupole density generated from the domain. The electrostatic energy due to the generation of spontaneous polarization and electric quadrupole density contributes to the formation of orientational deformation as the Frank elastic energy and spontaneous splay, respectively. Since the orientational configuration of the electric quadrupole density and in-plane spontaneous polarization is dependent on the molecular chirality, and the positive splay deformation of electric quadrupole density is induced by the spontaneous splay, the bending direction of in-plane spontaneous polarization depends on the chirality of constituent lipids. The electrostatic energy due to the in-plane spontaneous polarization is dependent on the orientational deformation of in-plane spontaneous polarization, and bends the domain shape towards the bending direction of the in-plane spontaneous polarization. It has been demonstrated that the chiral dependence of the domain shapes of lipid monolayers originated from the chiral dependence of orientational structure due to the electric quadrupole density. PMID- 17411169 TI - Estimating the influence of finite instrumental resolution on elastic neutron scattering intensities from proteins. AB - Recent experimental and simulation studies show that the fractional Ornstein Uhlenbeck process describes well the single particle motions in internal protein dynamics. Here the authors use this model to estimate the influence of finite instrumental resolution on elastic neutron scattering intensities from hydrated protein powders. They give, in particular, an estimation of the attenuation factor for the observed atomic position fluctuations, assuming a Gaussian and a triangular resolution function. PMID- 17411170 TI - Perspective: From field-ion microscopy of single atoms to atom-probe tomography: A journey: "Atom-probe tomography" [Rev. Sci. Instrum. 78, 031101 (2007)]. PMID- 17411171 TI - Invited review article: Atom probe tomography. AB - The technique of atom probe tomography (APT) is reviewed with an emphasis on illustrating what is possible with the technique both now and in the future. APT delivers the highest spatial resolution (sub-0.3-nm) three-dimensional compositional information of any microscopy technique. Recently, APT has changed dramatically with new hardware configurations that greatly simplify the technique and improve the rate of data acquisition. In addition, new methods have been developed to fabricate suitable specimens from new classes of materials. Applications of APT have expanded from structural metals and alloys to thin multilayer films on planar substrates, dielectric films, semiconducting structures and devices, and ceramic materials. This trend toward a broader range of materials and applications is likely to continue. PMID- 17411172 TI - Artificial submicron or nanometer speckle fabricating technique and electron microscope speckle photography. AB - In this article, a novel artificial submicro- or nanometer speckle fabricating technique is proposed by taking advantage of submicro or nanometer particles. In the technique, submicron or nanometer particles were adhered to an object surface by using ultrasonic dispersing technique. The particles on the object surface can be regarded as submicro or nanometer speckle by using a scanning electronic microscope at a special magnification. In addition, an electron microscope speckle photography (EMSP) method is developed to measure in-plane submicron or nanometer deformation of the object coated with the artificial submicro or nanometer speckles. The principle of artificial submicro or nanometer speckle fabricating technique and the EMSP method are discussed in detail in this article. Some typical applications of this method are offered. The experimental results verified that the artificial submicro or nanometer speckle fabricating technique and EMSP method is feasible. PMID- 17411173 TI - Quasi-Fourier-transform limited, scannable, high energy titanium-sapphire laser source for high resolution spectroscopy. AB - A tunable injection seeded Ti:sapphire laser source has been developed and tested. Slave-master tandem cavity and ramp-lock-and-fire concepts have been implemented and fully controlled by a digital signal processor. A Fourier transform-based analysis, as well as direct measurements, have demonstrated spectral linewidths in the range of 3.5-15 MHz (HWHM), with potential tunability over the entire Ti:sapphire lasing range. A quasi-Fourier-transform limited spectral linewidth is demonstrated assuming a secant hyperbolic shape of the electromagnetic field. Output energies >100 mJ have been reached with approximately 300 mJ of pump energy. The highest spectral purity is obtained using the quadruple pumping scheme. PMID- 17411174 TI - A highly integrated FPGA-based nuclear magnetic resonance spectrometer. AB - The digital circuits required for a nuclear magnetic resonance (NMR) spectrometer, including a pulse programmer, a direct digital synthesizer, a digital receiver, and a PC interface, have been built inside a single chip of the field-programmable gate-array (FPGA). By combining the FPGA chip with peripheral analog components, a compact, laptop-sized homebuilt spectrometer has been developed, which is capable of a rf output of up to 400 MHz with amplitude-, phase-, frequency-, and pulse-modulation. The number of rf channels is extendable up to three without further increase in size. PMID- 17411175 TI - Feedback system for improving the performance of extended x-ray absorption fine structure measurements. AB - A rapid fine energy tuning method for a double-crystal monochromator (DCM) is developed by combining a feedback control system with a phase sensitive detector. This method maintains both a parallel angle between the first and second crystals of the DCM and the peak of the rocking curve. The method is used to perform a continuous energy scan of the DCM without readjustment of the angle between the two crystals. The time taken by this method to measure the extended x-ray absorption fine structure is 60% of that taken by a conventional procedure. PMID- 17411176 TI - A precise method to determine the angular distribution of backscattered light to high angles. AB - We present an approach to measure the angular dependence of the diffusely scattered intensity of a multiple scattering sample in backscattering geometry. Increasing scattering strength give rise to an increased width of the coherent backscattering and sets higher demands on the angular detection range. This is of particular interest in the search for the transition to Anderson localization of light. To cover a range of -60 degrees to +85 degrees from direct back reflection, we introduced a new parallel intensity recording technique. This allows one-shot measurements, with fast alignment and short measuring time, which prevents the influence of illumination variations. Configurational average is achieved by rotating the sample and singly scattered light is suppressed with the use of circularly polarized light up to 97%. This implies that backscattering enhancements of almost two can be achieved. In combination with a standard setup for measuring small angles up to +/-3 degrees , a full characterization of the coherent backscattering cone can be achieved. With this setup we are able to accurately determine transport mean free paths as low as 235 nm. PMID- 17411177 TI - Dead-time optimized time-correlated photon counting instrument with synchronized, independent timing channels. AB - Time-correlated single photon counting is a powerful method for sensitive time resolved fluorescence measurements down to the single molecule level. The method is based on the precisely timed registration of single photons of a fluorescence signal. Historically, its primary goal was the determination of fluorescence lifetimes upon optical excitation by a short light pulse. This goal is still important today and therefore has a strong influence on instrument design. However, modifications and extensions of the early designs allow for the recovery of much more information from the detected photons and enable entirely new applications. Here, we present a new instrument that captures single photon events on multiple synchronized channels with picosecond resolution and over virtually unlimited time spans. This is achieved by means of crystal-locked time digitizers with high resolution and very short dead time. Subsequent event processing in programmable logic permits classical histogramming as well as time tagging of individual photons and their streaming to the host computer. Through the latter, any algorithms and methods for the analysis of fluorescence dynamics can be implemented either in real time or offline. Instrument test results from single molecule applications will be presented. PMID- 17411178 TI - Qualification of a fluorescence spectrometer for measuring true fluorescence spectra. AB - New analytical methods using fluorescence detection are becoming increasingly quantitative and require easy-to-use material standards for fluorometer qualification and method validation. NIST is responding to this need by developing and producing such standards. Reported here is the first step in this process, which is to qualify a research-grade fluorescence spectrometer for measuring true fluorescence spectra of reference material candidates. "True" spectra are defined here as those with fluorescence intensity, either relative or absolute as required, and wavelength both being reported with high accuracy and known precision, after wavelength has been calibrated and corrections for excitation intensity and detection system response have been applied. The uncertainties determined in relative and absolute intensity-corrected fluorescence spectra using both calibrated source (CS)- and calibrated detector (CD)-based methods were compared. The CS-based method gave uncertainties, typically about +/-5% for relative spectral correction, that were about half that of the CD-based method for determining both relative and absolute spectral correction factors. Absolute spectral correction factors can be determined using either method without knowing the optical geometry of the instrument. The absolute spectral correction factors were found to have much larger uncertainties than the corresponding relative correction factors with uncertainties for the CS based method of +/-10% to +/-15% being typical and +/-20% or more not being uncommon, particularly for excitation and emission wavelengths below 400 nm. Uncertainties arising from detection system nonlinearity and instrument polarization ratios were also explored. PMID- 17411179 TI - Mega-ampere submicrosecond generator GIT-32. AB - The GIT-32 current generator was developed for experiments with current carrying pulsed plasma. The main parts of the generator are capacitor bank, multichannel multigap spark switches, low inductive current driving lines, and central load part. The generator consists of four identical sections, connected in parallel to one load. The capacitor bank is assembled from 32 IEK-100-0.17 (0.17 microF, 40 nH, 100 kV) capacitors, connected in parallel. It stores approximately 18 kJ at 80 kV charging voltage. Each two capacitors are commuted to a load by a multigap spark switch with eight parallel channels. Switches operate in ambient air at atmospheric pressure. The GIT-32 generator was tested with 10, 15, and 20 nH inductive loads. At 10 nH load and 80 kV of charging voltage it provides 1 MA of current amplitude and 490 ns rise time with 0.8 Omega damping resistors in discharge circuit of each capacitor and 1.34 MA530 ns without resistors. The net generator inductance without a load was optimized to be as low as 12 nH, which results in extremely low self-impedance of the generator ( approximately 0.05 Omega). It ensures effective energy coupling with low impedance loads like Z pinch. The generator operates reliably without any adjustments in 40-80 kV range of charging voltage. Maximum jitter (relative to a triggering pulse) at 40 kV charging voltage is about 7 ns and lower at higher charging voltages. Operation and handling are very simple, because no oil and no purified gases are required for the generator. The GIT-32 generator has dimensions of 3200 x 3200 x 400 mm(3) and total weight of about 2500 kg, thus manifesting itself as a simple, robust, and cost effective apparatus. PMID- 17411180 TI - Langmuir probe system for dusty plasmas under microgravity. AB - This article describes a fully automated 2D-scanning Langmuir probe system for dusty plasmas under microgravity. The design combines necessary features such as random sampling, radio frequency compensation, and a compact mechanical design. The various aspects of the probe implementation and the contamination problem in the dusty plasma environment are discussed and the functionality of the system is demonstrated by measurements performed on parabolic flights. PMID- 17411181 TI - Main characteristics of the fast disruption mitigation valve. AB - The article presents a detailed investigation of the fast disruption mitigation valve developed at FZJ Juelich. The essence of this study is the novel direct observation of the piston motion by means of a fast framing camera. The piston stroke and the injection duration are shown to strongly depend on the operational pressure and the used gas. The same is true for the valve throughput. The analysis revealing the leading contribution of the injection duration in this modification is given. The knowledge of the injection duration is also used to reconstruct the characteristic pressure decay rates and the gas outflow rates. The means to increase the gas outflow are discussed. The main found valve characteristics are: (1) valve reaction time, i.e., the delay between the application of the trigger signal and the achievement of reliably observable opening 0.5 mm, is about 0.3 ms; (2) the maximum achieved throughput is 7.5 bar l for argon and 9.5 bar l for helium; (3) the maximum delivery rates are 500 bar l s(-1) for Ar and 1500 bar l s(-1) for He. PMID- 17411182 TI - Modeling magnetic fields measured by surface probes embedded in a cylindrical flux conserver. AB - Calculating magnetic fields at the surface of a flux conserver, perfect conductor, for displaced plasma currents is useful for understanding modes of a Z pinch. The magnetic fields measured at the flux conserver are a sum of the magnetic fields from the plasma current and the eddy currents which form in the walls to keep the flux constant. While the magnetic field at the wall from the plasma current alone is easily calculated using the Biot-Savart law, finding the eddy currents in the flux conserver which satisfy the boundary conditions can be a tedious process. A simple method of calculating the surface magnetic field for a given Z-pinch displacement off-axis is derived for a cylindrical flux conserver. This relationship does not require the explicit calculation of the eddy currents, saving time when analyzing surface magnetic probe measurements. Analytic expressions can be used to describe the surface magnetic field which increase the understanding of the magnetic probe measurements. PMID- 17411183 TI - Measurement of the D alpha spectrum produced by fast ions in DIII-D. AB - Fast ions are produced by neutral beam injection and ion cyclotron heating in toroidal magnetic fusion devices. As deuterium fast ions orbit around the device and pass through a neutral beam, some deuterons neutralize and emit D(alpha) light. For a favorable viewing geometry, the emission is Doppler shifted away from other bright interfering signals. In the 2005 campaign, we built a two channel charge-coupled device based diagnostic to measure the fast-ion velocity distribution and spatial profile under a wide variety of operating conditions. Fast-ion data are acquired with a time resolution of approximately 1 ms, spatial resolution of approximately 5 cm, and energy resolution of approximately 10 keV. Background subtraction and fitting techniques eliminate various contaminants in the spectrum. Neutral particle and neutron diagnostics corroborate the D(alpha) measurement. Examples of fast-ion slowing down and pitch angle scattering in quiescent plasma and fast-ion acceleration by high harmonic ion cyclotron heating are presented. PMID- 17411184 TI - Particle positioning techniques for dusty plasma experiments. AB - Video microscopy is a widely applied diagnostic to investigate the structure and the dynamics of particles in dusty plasmas. Reliable algorithms are required to accurately recover particle positions from the camera images. Here, four different particle positioning techniques have been tested on artificial and experimental data of dusty plasma situations. Two methods that rely on pixel intensity thresholds were found to be strongly affected by pixel-locking errors and by noise. Two other methods-one applying spatial bandpass filters and the other fitting polynomials to the intensity pattern-yield subpixel resolution under various conditions. These two methods have been shown to be ideally suited to recover particle positions even from small-scale fluctuations that are used to derive the normal mode spectra of finite dust clusters. PMID- 17411185 TI - Development of a microlateral force sensor and its evaluation using lateral force microscopy. AB - A microlateral force sensor (MLFS) was developed and evaluated using atomic force microscopy (AFM). The sensor was attached to a sensing table supported by a suspension system. The lateral motion of the sensing table was activated by a comb actuator. The driving voltage to the comb actuator was controlled to maintain a constant position of the sensing table by detecting the tunneling current at a detector, which consisted of two electrodes where the bias voltage was applied. An AFM was used to apply a lateral force to the sensing table of the sensor. When the probe of a cantilever was pressed against the sensing table and a raster scanning was conducted, the driving voltage of the comb actuator changed to compensate the friction force between the probe and sensing table. AFM measurements of an asperity array on the sensing table were conducted, and a lateral force microscopy image (LFM) was obtained from the change in driving voltage. The image by MLFS was very similar to the LFM image that was conventionally obtained from torsion of the cantilever. The LFM image strongly correlated with the gradient image calculated from the AFM topographic image. The force sensitivity of the MLFS was determined by comparing the LFM image obtained by using the MLFS with the tangential force derived from the gradient of the AFM image. PMID- 17411186 TI - Compact flashlamp-based fluorescence imager for use under ambient-light conditions. AB - A low-power, lightweight, multiwavelength fluorescence imager based on the use of a compact xenon flashlamp, bandpass filters, gated charge-coupled device camera, and digital image processing was developed for use on an autonomous rover vehicle. The imager discriminates against ambient light by use of microsecond excitation pulses along with synchronized camera operation to limit the time period in which ambient-light photocounts are accumulated, and digital image subtraction to remove background counts. In a 10 cm square field of view, weak fluorescence, equivalent to 0.05 pmol fluorescein/mm(2), can be quantified against a white-light background equivalent to shaded sunlight. For application in autonomous search for organisms in extreme environments such as in situ desert rock or soil, the instrument was equipped with a set of fluorescence excitation filters (380, 450, 545, and 600 nm) and emission filters (460, 510, 620, and 740 nm) suitable for detection of chlorophyll, applied stains for protein, DNA, lipid and carbohydrate, and autofluorescence. True-color images were obtained through red-green-blue imaging filters (630, 535, and 470 nm) used with white-light flashes. Automated focusing on ground features was based on the R-band image and was carried out prior to fluorescence image acquisition. PMID- 17411187 TI - In situ azimuthal rotation device for linear dichroism measurements in scanning transmission x-ray microscopy. AB - A novel miniature rotation device used in conjunction with a scanning transmission x-ray microscope is described. It provides convenient in situ sample rotation to enable measurements of linear dichroism at high spatial resolution. The design, fabrication, and mechanical characterization are presented. This device has been used to generate quantitative maps of the spatial distribution of the orientation of proteins in several different spider and silkworm silks. Specifically, quantitative maps of the dichroic signal at the C 1s-->pi* (amide) transition in longitudinal sections of the silk fibers give information about the spatial orientation, degree of alignment, and spatial distribution of protein peptide bonds. A new approach for analyzing the dichroic signal to extract orientation distributions, in addition to magnitudes of aligned components, is presented and illustrated with results from Nephila clavipes dragline spider silk measured using the in situ rotation device. PMID- 17411188 TI - X-ray imaging with grazing-incidence microscopes developed for the LIL program. AB - This article describes x-ray imaging with grazing-incidence microscopes, developed for the experimental program carried out on the Ligne d'Integration Laser (LIL) facility [J. P. Le Breton et al., Inertial Fusion Sciences and Applications 2001 (Elsevier, Paris, 2002), pp. 856-862] (24 kJ, UV-0.35 nm). The design includes a large target-to-microscope (400-700 mm) distance required by the x-ray ablation issues anticipated on the Laser MegaJoule facility [P. A. Holstein et al., Laser Part. Beams 17, 403 (1999)] (1.8 MJ) which is under construction. Two eight-image Kirkpatrick-Baez microscopes [P. Kirkpatrick and A. V. Baez J. Opt. Soc. Am. 38, 766 (1948)] with different spectral wavelength ranges and with a 400 mm source-to-mirror distance image the target on a custom built framing camera (time resolution of approximately 80 ps). The soft x-ray version microscope is sensitive below 1 keV and its spatial resolution is better than 30 microm over a 2-mm-diam region. The hard x-ray version microscope has a 10 microm resolution over an 800-microm-diam region and is sensitive in the 1-5 keV energy range. Two other x-ray microscopes based on an association of toroidal/spherical surfaces (T/S microscopes) produce an image on a streak camera with a spatial resolution better than 30 microm over a 3 mm field of view in the direction of the camera slit. Both microscopes have been designed to have, respectively, a maximum sensitivity in the 0.1-1 and 1-5 keV energy range. We present the original design of these four microscopes and their test on a dc x ray tube in the laboratory. The diagnostics were successfully used on LIL first experiments early in 2005. Results of soft x-ray imaging of a radiative jet during conical shaped laser interaction are shown. PMID- 17411189 TI - Implementation of diffractive optical element in four-wave mixing scheme for ex situ characterization of hydride vapor phase epitaxy-grown GaN layers. AB - A holographic beam splitter has been integrated into a picosecond four-wave mixing (FWM) scheme. This modification significantly simplified the procedure of dynamic grating recording, thus making the FWM technique an easy-to-use tool for the holographic characterization of wide band gap materials. The novel FWM scheme was applied for characterization of hydride vapor phase epitaxy-grown undoped GaN layers of different thickness. It allowed the determination of carrier lifetime, diffusion coefficient, and carrier diffusion length by optical means, as well as the study of carrier recombination peculiarities with respect to dislocation and excess carrier density. PMID- 17411190 TI - Computer-controlled susceptometer for investigating the linear and nonlinear dielectric response. AB - A fully automatized alternating current (ac) susceptometer is constructed for simultaneous measurements of the phase resolved complex linear and complex nonlinear ac susceptibilities of lossy and dispersive dielectric materials. This relatively simple setup allows measurements over a wide range of experimental variables, such as ac amplitudes up to 40 V, frequencies from 10(-2) to 10(3) Hz, and temperatures from 100 to 600 K utilizing only current/voltage and analog/digital converters and a personal computer. In contrast with the commonly used analysis of the charge accumulated on a standard capacitor in series with the sample our method is based on the analysis of the current flowing directly through the sample. Absence of any capacitive voltage dividers in the measurement circuit eliminates uncontrolled phase shifts. This is why the instrument provides high quality, nonlinear susceptibility data and in particular appears as a very convenient tool for discrimination between continuous and discontinuous phase transitions when determining the sign of the real part of the third order dielectric susceptibility. PMID- 17411191 TI - Portable chamber for the study of UHV prepared electrochemical interfaces by hard x-ray diffraction. AB - We report on a new electrochemical cell setup, combined with a portable UHV chamber, for in situ x-ray diffraction using synchrotron radiation. In contrast to more traditional electrochemical sample preparation schemes, atomically clean and well-ordered surfaces are routinely prepared by UHV methods, even in the case of reactive elements or alloys. Samples can be transferred from larger UHV systems into the portable chamber without exposure to ambient air. They can then be studied successively in UHV, in controlled gas atmospheres, and in contact with electrolyte solutions under applied electrochemical potential. The electrochemical setup employs a droplet geometry, which guarantees good electrochemical conditions during in situ x-ray measurements combined with voltammetry. We present first experimental results of Cu deposition on GaAs(001) and on freshly produced nanometric Pd(001) islands on Cu(0.83)Pd(0.17)(001), respectively. PMID- 17411192 TI - Low-temperature differential-thermal analysis to measure variations in entropy. AB - We describe how we can precisely measure variations in the entropy S of small solid samples below room temperature, as a function the temperature T or the external magnetic field H, respectively. A simple differential-thermal analysis technique allows, in principle, for the measurement of variations in S without any instrumental broadening of the data. The method is particularly well suited for the detection of sharp phase transitions in samples of milligram size. PMID- 17411193 TI - An instrument to investigate femtochemistry on metal surfaces in real space. AB - A newly established combination of a femtosecond laser with a low temperature scanning tunneling microscope is described, which facilitates one to analyze femtochemistry on metal surfaces in real space. The combined instrument enables focusing the laser to some tens of micrometers and guiding it reproducibly into the tunneling gap with the aid of in situ movable mirrors. Furthermore, a method to determine the focus size on the sample is presented. The focus size is used to calculate the electron and phonon temperatures at the surface. Despite the additional noise introduced by laser operation the vertical resolution of the microscope lies below 1 pm. The potential of the instrument is demonstrated on para-chloronitrobenzene clusters adsorbed on Au(111). Single chloronitrobenzene molecules diffuse upon femtosecond laser irradiation; some smaller clusters rotate by multiples of 30 degrees ; clusters of less compact form rearrange to close-packed clusters. PMID- 17411194 TI - Gravitational capillary viscometer for low-temperature liquids. AB - A pressurized gravitational capillary viscometer was developed for subcooled liquefied gases such as oxygen, nitrogen, hydrogen, and methane. It acquires accurate absolute viscosity measurements with an uncertainty of 1% at a 95.5% confidence level, necessary for the demands of aerospace engineering. The viscometer includes a coiled capillary of electroformed nickel that hydraulically connects two reservoirs located at equal heights. Using helium gas to temporarily drive liquid from a third supply reservoir, a level difference is created between the first two reservoirs without the need to tilt or invert the viscometer. Helium gas is then used to pressurize the first two reservoirs equally. Each reservoir holds a capacitive liquid-level sensor that both measure the flow induced by gravity through the capillary more precisely than by visual observations through windows. Viscosity data for liquid oxygen were acquired in the pressure domain from 0.15 to 1.0 MPa and in the temperature domain from the normal boiling point to near the freezing point. Published viscosity data and correlations for subcooled liquid oxygen are not precise nor complete in this density regime. Accordingly, the viscosity data delivered by the viscometer of the present work are superior. PMID- 17411195 TI - Circular Couette cell for two-dimensional fluid dynamics experiments. AB - A novel experiment to investigate fluid dynamics in quasi-two-dimensional flows has been built. A soap film is suspended horizontally in an annular channel with a rotating outer boundary, providing mean flow shear, and a vortex array is forced electromagnetically. The experiment will investigate sheared flow stability and the effect of mean flow shear on local vorticity and coherent structures. Particle image velocimetry measurements demonstrate the production of mean flow shear and induced vortices. PMID- 17411196 TI - Shock tube/time-of-flight mass spectrometer for high temperature kinetic studies. AB - A shock tube (ST) with online, time-of-flight mass spectrometric (TOF-MS) detection has been constructed for the study of elementary reactions at high temperature. The ST and TOF-MS are coupled by a differentially pumped molecular beam sampling interface, which ensures that the samples entering the TOF-MS are not contaminated by gases drawn from the cold end wall thermal boundary layer in the ST. Additionally, the interface allows a large range of postshock pressures to be used in the shock tube while maintaining high vacuum in the TOF-MS. The apparatus and the details of the sampling system are described along with an analysis in which cooling of the sampled gases and minimization of thermal boundary layer effects are discussed. The accuracy of kinetic measurements made with the apparatus has been tested by investigating the thermal unimolecular dissociation of cyclohexene to ethylene and 1,3-butadiene, a well characterized reaction for which considerable literature data that are in good agreement exist. The experiments were performed at nominal reflected shock wave pressures of 600 and 1300 Torr, and temperatures ranging from 1260 to 1430 K. The rate coefficients obtained are compared with the earlier shock tube studies and are found to be in very good agreement. As expected no significant difference is observed in the rate constant between pressures of 600 and 1300 Torr. PMID- 17411197 TI - A compact setup to study homogeneous nucleation and condensation. AB - An experiment is presented to study homogeneous nucleation and the subsequent droplet growth at high temperatures and high pressures in a compact setup that does not use moving parts. Nucleation and condensation are induced in an adiabatic, stationary expansion of the vapor and an inert carrier gas through a Laval nozzle. The adiabatic expansion is driven against atmospheric pressure by pressurized inert gas its mass flow carefully controlled. This allows us to avoid large pumps or vacuum storage tanks. Because we eventually want to study the homogeneous nucleation and condensation of zinc, the use of carefully chosen materials is required that can withstand pressures of up to 10(6) Pa resulting from mass flow rates of up to 600 l(N) min(-1) and temperatures up to 1200 K in the presence of highly corrosive zinc vapor. To observe the formation of droplets a laser beam propagates along the axis of the nozzle and the light scattered by the droplets is detected perpendicularly to the nozzle axis. An ICCD camera allows to record the scattered light through fused silica windows in the diverging part of the nozzle spatially resolved and to detect nucleation and condensation coherently in a single exposure. For the data analysis, a model is needed to describe the isentropic core part of the flow along the nozzle axis. The model must incorporate the laws of fluid dynamics, the nucleation and condensation process, and has to predict the size distribution of the particles created (PSD) at every position along the nozzle axis. Assuming Rayleigh scattering, the intensity of the scattered light can then be calculated from the second moment of the PSD. PMID- 17411198 TI - Time-of-flight mass spectrometry for time-resolved measurements. AB - A time-resolved time-of-flight mass spectrometer (TOF-MS) that can simultaneously monitor multiple species on the millisecond time scale has been constructed. A pulsed photolysis laser is used to initiate reaction, and then via a pinhole the reaction mixture is sampled by the TOF-MS. The ions are created by photoionization via either a discharge lamp or a pulsed laser. Comparison between the two ionization sources showed that the laser is at least an order of magnitude more efficient, based on the time to accumulate the data. Also, unlike the continuous lamp the pulsed laser is not mass limited. Frequency tripling the 355 nm output of a Nd:YAG laser provided a convenient laser ionization source. However, using a dye laser provided an equally intense laser ionization source with the ability to tune the vacuum ultraviolet (vuv) light. To show the versatility of the system the kinetics of the reaction of SO and ClSO radicals with NO(2) were simultaneously measured, and using the dye laser the vuv light was tuned to 114 nm in order to observe H(2)CO being formed from the reaction between CH(3)CO and O(2). PMID- 17411199 TI - Method for measurement of the density of thin films of small organic molecules. AB - An accurate and sensitive method is reported to measure the thin-film density of vacuum-deposited, small-molecular organic semiconductor materials. A spectrophotometer and surface profiler had been used to determine the mass and thickness of organic thin film, respectively. The calculated density of tris-(8 hydroxyquinolato) aluminum (Alq(3)) thin film was 1.31+/-0.01 gcm(3). Vacuum pressures and thin-film growth rates are found to have less impact on the thin film density of organic material. However, the thin-film density of organic material strongly depends on its chemical structure and molecular weight. Specifically, the chemical structure determines the density of organic material that affects the molecular volume and intermolecular stacking. PMID- 17411200 TI - Internal-energy measurements of angle-resolved product CO2 in catalytic CO oxidation by means of infrared chemiluminescence. AB - Measurements of both vibrational and rotational energies of product CO(2) in CO oxidation on palladium surfaces have been successfully performed as a function of the desorption angle by means of infrared chemiluminescence. The remarkable angle dependences of both energies indicate facile energy partitioning in repulsive desorption and provide new dimensions in the study of surface reaction dynamics as well as additional insights into the product formation site. Details of the apparatus for energy analysis of angle-resolved products are described, especially on how to pick up extremely weak infrared emission signals. PMID- 17411201 TI - Micropatterned silicone elastomer substrates for high resolution analysis of cellular force patterns. AB - Cellular forces are closely related to many physiological processes, including cell migration, growth, division, and differentiation. Here, we describe newly developed techniques to measure these forces with high spatial resolution. Our approach is based on ultrasoft silicone elastomer films with a regular microstructure molded into the surface. Mechanical forces applied by living cells to such films result in elastomer deformation which can be quantified by video microscopy and digital image processing. From this deformation field forces can be calculated. Here we give detailed accounts of the following issues: (1) the preparation of silicon wafers as molds for the microstructures, (2) the fabrication of microstructured elastomer substrates, (3) the in-depth characterization of the mechanical properties of these elastomers, (4) the image processing algorithms for the extraction of cellular deformation fields, and (5) the generalized first moment tensor as a robust mathematical tool to characterize whole cell activity. We present prototype experiments on living myocytes as well as on cardiac fibroblasts and discuss the characteristics and performance of our force measurement technique. PMID- 17411202 TI - Motion analysis of grip and release with fingers using simple magnetic detection system. AB - We have developed a magnetic grip-and-release motion detection system to measure time differences among fingers during motion. We designed a magnetic sensing system consisting of a magnetic oscillation coil, sensing coil, and circuit unit. We measured wave forms of grip-and-release motion (15 s) of seven healthy volunteers using the system. To provide the grip-and-release timing of each finger, we used the reference voltage of each subject in the state of grasping a rod with a 30 mm diameter. Using the reference voltage, the time differences in the finger movements of all volunteers were detected. The detected grip-and release time difference of both dominant and nondominant hands had a main tendency in which the closing movement of the little finger is fast, within 10 ms, and the opening movement is slow, within 13 ms. Our data suggest that the new magnetic sensing system has the potential to detect the quantitative value of the time difference in grip-and-release motion among fingers. PMID- 17411203 TI - Gamma-ray detection efficiency of the microchannel plate installed as an ion detector in the low energy particle instrument onboard the GEOTAIL satellite. AB - A microchannel plate (MCP) assembly has been used as an ion detector in the low energy particle (LEP) instrument onboard the magnetospheric satellite GEOTAIL. Recently the MCP assembly has detected gamma rays emitted from an astronomical object and has been shown to provide unique information of gamma rays if they are intense enough. However, the detection efficiency for gamma rays was not measured before launch, and therefore we could not analyze the LEP data quantitatively. In this article, we report the gamma-ray detection efficiency of the MCP assembly. The measured efficiencies are 1.29%+/-0.71% and 0.21%+/-0.14% for normal incidence 60 and 662 keV gamma rays, respectively. The incident angle dependence is also presented. Our calibration is crucial to study high energy astrophysical phenomena by using the LEP. PMID- 17411204 TI - Multipurpose EPR loop-gap resonator and cylindrical TE011 cavity for aqueous samples at 94 GHz. AB - A loop-gap resonator (LGR) and a cylindrical TE(011) cavity resonator for use at W band, 94 GHz, have been designed and characterized using the Ansoft (Pittsburgh, PA) high frequency structure simulator (HFSS; Version 10.0). Field modulation penetration was analyzed using Ansoft MAXWELL 3D (Version 11.0). Optimizing both resonators to the same sample sizes shows that EPR signal intensities of the LGR and TE(011) are similar. The 3 dB bandwidth of the LGR, on the order of 1 GHz, is a new advantage for high frequency experiments. Ultraprecision electric discharge machining (EDM) was used to fabricate the resonators from silver. The TE(011) cavity has slots that are cut into the body to allow penetration of 100 kHz field modulation. The resonator body is embedded in graphite, also cut by EDM techniques, for a combination of reasons that include (i) reduced microwave leakage and improved TE(011) mode purity, (ii) field modulation penetration, (iii) structural support for the cavity body, and (iv) machinability by EDM. Both resonators use a slotted iris. Variable coupling is provided by a three-stub tuning element. A collet system designed to hold sample tubes has been implemented, increasing repeatability of sample placement and reducing sample vibration noise. Initial results include multiquantum experiments up to 9Q using the LGR to examine 1 mM 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO) in aqueous solution at room temperature and field modulation experiments using the TE(011) cavity to obtain an EPR spectrum of 1 microM TEMPO. PMID- 17411205 TI - Analysis of a modular generator for high-voltage, high-frequency pulsed applications, using low voltage semiconductors (< 1 kV) and series connected step up (1:10) transformers. AB - This article discusses the operation of a modular generator topology, which has been developed for high-frequency (kHz), high-voltage (kV) pulsed applications. The proposed generator uses individual modules, each one consisting of a pulse circuit based on a modified forward converter, which takes advantage of the required low duty cycle to operate with a low voltage clamp reset circuit for the step-up transformer. This reduces the maximum voltage on the semiconductor devices of both primary and secondary transformer sides. The secondary winding of each step-up transformer is series connected, delivering a fraction of the total voltage. Each individual pulsed module is supplied via an isolation transformer. The assembled modular laboratorial prototype, with three 5 kV modules, 800 V semiconductor switches, and 1:10 step-up transformers, has 80% efficiency, and is capable of delivering, into resistive loads, -15 kV1 A pulses with 5 micros width, 10 kHz repetition rate, with less than 1 micros pulse rise time. Experimental results for resistive loads are presented and discussed. PMID- 17411206 TI - Noise-rejection techniques for impedance and dielectric spectrometers using ubiquitous test and measurement equipment. AB - This work encompassed the development for a frequency-domain impedance and dielectric spectrometer using ubiquitous test and measurement equipment, i.e., signal generators and digital oscilloscopes. Various methods of amplification, noise rejection, and computations were employed to achieve the desired goals. The frequency range of 100 mHz-1 MHz was tested using air capacitors of 3.7 and 14.5 pF and an applied voltage range of 10-300 mV. The multichannel instrument produced a stable and reproducible dual-phase (real and imaginary or magnitude and phase) current sensitivity of 250 fA with an average phase stability of less than 0.5 degrees (tan delta<10(-2)) and a single-phase (magnitude only) current sensitivity of 60 fA. PMID- 17411207 TI - Four-probe measurements of the in-plane thermoelectric properties of nanofilms. AB - Measuring in-plane thermoelectric properties of submicron thin films has remained a challenging task. Here we report a method based on a suspended microdevice for four-probe measurements of the Seebeck coefficient, thermal conductivity, electrical conductivity, and thermoelectric figure of merit of patterned indium arsenide (InAs) nanofilms assembled on the microdevice. The contact thermal resistance and intrinsic thermal resistance of the 40 nm thick InAs nanofilm sample were measured by using the nanofilm itself as a differential thermocouple to determine the temperature drops at the contacts. The microdevice was also used to measure a 190 nm thick silicon dioxide (SiO(2)) film and the results were compared with those reported in the literature. A through-substrate hole under the suspended microdevice allows for transmission electron microscopy characterization of the nanofilm sample assembled on the device. This capability enables one to correlate the measured thermoelectric properties with the crystal structures of the nanofilm. PMID- 17411208 TI - Near-infrared thermography using a charge-coupled device camera: application to microsystems. AB - Using near-infrared thermography microscopy and a low-cost charge-coupled device (CCD) camera, we have designed a system which is able to deliver quantitative submicronic thermal images. Using a theoretical model based on Planck's law and CCD sensor properties allowed us to determine a minimal theoretical detection temperature and an optimal temperature sensitivity of our system. In order to validate this method, we show a good relationship between a theoretical study and a thermal measurement of a microsample. PMID- 17411209 TI - Streaked optical pyrometer system for laser-driven shock-wave experiments on OMEGA. AB - The temperature of laser-driven shock waves is of interest to inertial confinement fusion and high-energy-density physics. We report on a streaked optical pyrometer that measures the self-emission of laser-driven shocks simultaneously with a velocity interferometer system for any reflector (VISAR). Together these diagnostics are used to obtain the temporally and spatially resolved temperatures of approximately megabar shocks driven by the OMEGA laser. We provide a brief description of the diagnostic and how it is used with VISAR. Key spectral calibration results are discussed and important characteristics of the recording system are presented. PMID- 17411210 TI - Differential photoacoustic cell for electrochemical and dynamic process with temperature control. AB - A novel differential photoacoustic cell (DPC) for the study of dynamical processes has been developed. The DPC has the capability to measure in real time the amplitude and phase signals for the reference and the sample under study. The simultaneous measurement of both signals eliminates the instrumental function, and the presence of noise, due to any deviation originated by electrical, optical, and environmental factors. The DPC can be used at different temperature profiles in order to obtain the instrumental function IF(t,T). The DPC also has all the elements of an electrochemical cell capable of following the electrochemical processes. As a result of this new instrumentation it is possible to obtain in real time the amplitude and phase signals coming from the sample without any interference from the system and the viability to monitor in situ electrochemical and thermal processes. Two cases are presented as an illustrative demonstration of work fields: the electrodeposition of zinc on a steel substrate as well as the study of water and calcium ion diffusion into organic layers. PMID- 17411211 TI - Magnetic compound refractive lens for focusing and polarizing cold neutron beams. AB - Biconcave cylindrical lenses are used to focus beams of x rays or neutrons using the refractive properties of matter. In the case of neutrons, the refractive properties of magnetic induction can similarly focus and simultaneously polarize the neutron beam without the concomitant attenuation of matter. This concept of a magnetic refractive lens was tested using a compound lens consisting of 99 pairs of cylindrical permanent magnets. The assembly successfully focused the intensity of a white beam of cold neutrons of one spin state at the detector, while defocusing the other. This experiment confirmed that a lens of this nature may boost the intensity locally by almost an order of magnitude and create a polarized beam. An estimate of the performance of a more practically dimensioned device suitable for incorporation in reflectometers and slit-geometry small angle scattering instruments is given. PMID- 17411212 TI - A lead-free piezoelectric transformer in radial vibration modes. AB - In this study, a disk-shaped piezoelectric transformer was fabricated using lead free (K,Na)NbO(3)-based ceramics with high mechanical quality factor. The transformer can operate in the fundamental or the third radial vibration mode. The transformer is poled along the thickness direction. The top surface is covered by ring/dot silver electrodes separated by an annular gap which serve as the input and output parts of the transformer, respectively. The bottom surface, fully covered with a silver electrode, is grounded as a common electrode. The dimensions of the top ring/dot electrodes are designed such that the third radial vibration mode can be strongly excited. The electrical properties of the transformer with diameter of 34.2 mm and thickness of 1.9 mm were measured. For a temperature rise of 35 degrees C, the transformer has a maximum output power of 12 W. With the matching load, its maximum efficiency is >95%, and maximum voltage gains are 6.5 and 3.9 for the fundamental and the third radial vibration modes, respectively. It has potential to be used in power supply units and other electronic circuits. PMID- 17411213 TI - High performance temperature controlled UHV sample holder. AB - A requirement of many surface science studies is the capability to alter a sample temperature in a controlled mode. Sample preparation procedures such as heating or cooling ramps, high temperature spikes, fast annealing, or simply maintaining a sample at a very high, or very low, temperature are common. To address these issues, we describe the design and the construction of a multipurpose sample holder. Key points of this design are operation in an extended temperature range from liquid nitrogen (LN(2)) temperature to approximately 1300 K, temperature control during heating and cooling, low thermal inertia with rates up to 50 K s( 1) (heating) and -20 K s(-1) (cooling), and small heated volume to minimize background problems in thermal desorption spectroscopy (TDS) spectra. With this design the sample can be flash heated from LN(2) temperature to 1300 K and cooled down again in less than 100 s. This sample holder was mounted and tested in a multitechnique apparatus and adds a large number of sample preparation procedures as well as TDS to the list of already available surface analysis techniques. PMID- 17411214 TI - Use of quantum-point-contact high-electron-mobility-transistors for time domain multiplexing of large arrays of high impedance low temperature bolometers. AB - The use of a multiplexing readout for an array of bolometers simplifies the electronics and wiring, so making the readout of large arrays of bolometers (>100) feasible. Here we describe a time domain multiplexing technique and its performance based on the use of quantum-point-contact high-electron-mobility transistors as low temperature (to approximately 100 mK) switches for measuring high impedance (5...70 MOmega) resistances and sensors. The presented system is well matched to ground based millimetric astronomy demands. PMID- 17411215 TI - Design and performance of a sub-nanoradian resolution autocollimating optical lever. AB - Precision goniometry using optics has the advantage that it does not impose much stress on the object of investigation and, as such, is adopted extensively in gravitational wave detection, in torsion balances investigating fundamental forces, in specialized studies of biological samples, and it has potential applications in condensed matter physics. In this article we present the considerations that go into designing optical levers and discuss the performance of the instrument we have constructed. We motivate the design by considering an idealized setup and the limitations to the angular resolution induced by statistical fluctuations of the photon count rate and diffraction at the apertures. The effects of digitization of the count rate and of the spatial location of the photons on the image plane motivating the actual design are discussed next. Based on these considerations, we have developed an autocollimating optical lever which has a very high resolution and dynamic range. An array of 110 slits, of 90 microm width and a pitch of 182 microm, is located in the focal plane of a field lens, of focal length 1000 mm, and is illuminated by a CCFL tube. This array is imaged back onto the focal plane after retroreflection from a mirror placed just beyond the lens. The image is recorded on a linear charge-coupled device array at the rate of 1000 images/s and is processed through a special algorithm to obtain the centroid. The instrument has a centroid stability of approximately 3 x 10(-10) rad Hz(-1/2) and a dynamic range of approximately 10(7). PMID- 17411216 TI - Demagnetization of magnetically shielded rooms. AB - Magnetically shielded rooms for specific high resolution physiological measurements exploiting the magnetic field, e.g., of the brain (dc magnetoencephalography), low-field NMR, or magnetic marker monitoring, need to be reproducibly demagnetized to achieve reliable measurement conditions. We propose a theoretical, experimental, and instrumental base whereupon the parameters which affect the quality of the demagnetization process are described and how they have to be handled. It is demonstrated how conventional demagnetization equipment could be improved to achieve reproducible conditions. The interrelations between the residual field and the variability at the end of the demagnetization process are explained on the basis of the physics of ferromagnetism and our theoretical predictions are evaluated experimentally. PMID- 17411217 TI - Ultrafast time-domain spectroscopy based on high-speed asynchronous optical sampling. AB - High-speed asynchronous optical sampling (ASOPS) is a novel technique for ultrafast time-domain spectroscopy (TDS). It employs two mode-locked femtosecond oscillators operating at a fixed repetition frequency difference as sources of pump and probe pulses. We present a system where the 1 GHz pulse repetition frequencies of two Ti:sapphire oscillators are linked at an offset of Deltaf(R)=10 kHz. As a result, their relative time delay is repetitively ramped from zero to 1 ns within a scan time of 100 micros. Mechanical delay scanners common to conventional TDS systems are eliminated, thus systematic errors due to beam pointing instabilities and spot size variations are avoided when long time delays are scanned. Owing to the multikilohertz scan-rate, high-speed ASOPS permits data acquisition speeds impossible with conventional schemes. Within only 1 s of data acquisition time, a signal resolution of 6 x 10(-7) is achieved for optical pump-probe spectroscopy over a time-delay window of 1 ns. When applied to terahertz TDS, the same acquisition time yields high-resolution terahertz spectra with 37 dB signal-to-noise ratio under nitrogen purging of the spectrometer. Spectra with 57 dB are obtained within 2 min. A new approach to perform the offset lock between the two femtosecond oscillators in a master-slave configuration using a frequency shifter at the third harmonic of the pulse repetition frequency is employed. This approach permits an unprecedented time delay resolution of better than 160 fs. High-speed ASOPS provides the functionality of an all-optical oscilloscope with a bandwidth in excess of 3000 GHz and with 1 GHz frequency resolution. PMID- 17411218 TI - Compact optical fiber sensor smart node. AB - We present a new optical fiber sensor interrogator specifically designed for an embedded instrumentation system. The proposed system consists of a super luminescent diode as a broadband source, a high speed tunable micro-electro mechanical system (MEMS) filter, photodetector, and an integrated microprocessor for data aggregation, processing, and communication. The entire system is integrated together in a compact package to create a fiber "smart" sensor. The system is capable of interrogating a variety of multiplexed fiber sensors, processing the data, and communicating the results digitally. As an example, the system has been calibrated with an array of fiber Bragg grating sensors. PMID- 17411219 TI - BARCOS, an automation and remote control system for atmospheric observations with a Bruker interferometer. AB - In order to make long-term monitoring of the atmospheric composition using commercial Bruker Fourier transform spectrometers more cost effective, a system called BARCOS has been developed. The system enables one to perform the operation of the spectrometric atmospheric observations in a remotely controlled or autonomous way, without human presence at the measuring site. Several observation geometries are foreseen, including solar and lunar absorption spectrometry. BARCOS is built using existing commercial hardware and software components, including the Bruker software for the operation of the spectrometer (OPUS) and runs in a personal computer (Microsoft) environment. It includes a small meteorological station. It is a flexible system, allowing manual interventions at any time. To run BARCOS effectively, the only prerequisite is that internet access is available at the site of operation. This article describes the BARCOS system hardware and software configurations. PMID- 17411220 TI - Noncontact measurement of electrostatic fields: verification of modeled potentials within ion mobility spectrometer drift tube designs. AB - The heart of an ion mobility spectrometer is the drift region where ion separation occurs. While the electrostatic potentials within a drift tube design can be modeled, no method for independently validating the electrostatic field has previously been reported. Two basic drift tube designs were modeled using SIMION 7.0 to reveal the expected electrostatic fields: (1) A traditional alternating set of electrodes and insulators and (2) a truly linear drift tube. One version of the alternating electrode/insulator drift tube and two versions of linear drift tubes were then fabricated. The stacked alternating electrodes/insulators were connected through a resistor network to generate the electrostatic gradient in the drift tube. The two linear drift tube designs consisted of two types of resistive drift tubes with one tube consisting of a resistive coating within an insulating tube and the other tube composed of resistive ferrites. The electrostatic fields within each type of drift tube were then evaluated by a noncontact method using a Kelvin-Zisman type electrostatic voltmeter and probe (results for alternative measurement methods provided in supplementary material). The experimental results were then compared with the electrostatic fields predicted by SIMION. Both the modeling and experimental measurements reveal that the electrostatic fields within a stacked ion mobility spectrometer drift tube are only pseudo-linear, while the electrostatic fields within a resistive drift tube approach perfect linearity. PMID- 17411221 TI - High speed single charge coupled device Cranz-Schardin camera. AB - This article describes an ultrahigh speed visualization system based on a miniaturization of the Cranz-Schardin principle. It uses a set of high power light emitting diodes (LEDs) (Golden Dragon) as the light source and a highly sensitive charge coupled device (CCD) camera for reception. Each LED is fired in sequence and images the refraction index variation between two relay lenses, on a partial region of a CCD image sensor. The originality of this system consists in achieving several images on a single CCD during a frame time. The number of images is 4. The time interval between successive firings determines the speed of the imaging system. This time lies from 100 ns to 10 micros. The light pulse duration lies from 100 ns to 10 micros. The principle and the optical and electronic parts of such a system are described. As an example, some images of acoustic waves propagating in water are presented. PMID- 17411222 TI - Mass-controlled capillary viscometer for a Newtonian liquid: viscosity of water at different temperatures. AB - The operation principle of the mass-controlled capillary viscometer is presented for a Newtonian liquid. The derived equation for the temporal changes of the mass in a liquid column draining under gravity through a discharge capillary tube accounts self-consistently for the inertial convective term associated with the acceleration effect. The viscosity of water measured at different temperatures using the new approach is in good agreement with literature data. PMID- 17411223 TI - Precision instrumentation for rolling element bearing characterization. AB - This article describes an instrument to measure the error motion of rolling element bearings. This challenge is met by simultaneously satisfying four requirements. First, an axial preload must be applied to seat the rolling elements in the bearing races. Second, one of the races must spin under the influence of an applied torque. Third, rotation of the remaining race must be prevented in a way that leaves the radial, axial/face, and tilt displacements free to move. Finally, the bearing must be fixtured and measured without introducing off-axis loading or other distorting influences. In the design presented here, an air bearing reference spindle with error motion of less than 10 nm rotates the inner race of the bearing under test. Noninfluencing couplings are used to prevent rotation of the bearing outer race and apply an axial preload without distorting the bearing or influencing the measurement. Capacitive displacement sensors with 2 nm resolution target the nonrotating outer race. The error motion measurement repeatability is shown to be less than 25 nm. The article closes with a discussion of how the instrument may be used to gather data with sufficient resolution to accurately estimate the contact angle of deep groove ball bearings. PMID- 17411224 TI - Idealized gas turbine combustor for performance research and validation of large eddy simulations. AB - This paper details the design of a premixed, swirl-stabilized combustor that was designed and built for the express purpose of obtaining validation-quality data for the development of large eddy simulations (LES) of gas turbine combustors. The combustor features nonambiguous boundary conditions, a geometrically simple design that retains the essential fluid dynamics and thermochemical processes that occur in actual gas turbine combustors, and unrestrictive access for laser and optical diagnostic measurements. After discussing the design detail, a preliminary investigation of the performance and operating envelope of the combustor is presented. With the combustor operating on premixed methane/air, both the equivalence ratio and the inlet velocity were systematically varied and the flame structure was recorded via digital photography. Interesting lean flame blowout and resonance characteristics were observed. In addition, the combustor exhibited a large region of stable, acoustically clean combustion that is suitable for preliminary validation of LES models. PMID- 17411225 TI - Multipole shimming of permanent magnets using harmonic corrector rings. AB - Shimming systems are required to provide sufficient field homogeneity for high resolution nuclear magnetic resonance (NMR). In certain specialized applications, such as rotating-field NMR and mobile ex situ NMR, permanent magnet-based shimming systems can provide considerable advantages. We present a simple two dimensional shimming method based on harmonic corrector rings which can provide arbitrary multipole order shimming corrections. Results demonstrate, for example, that quadrupolar order shimming improves the linewidth by up to an order of magnitude. An additional order of magnitude reduction is in principle achievable by utilizing this shimming method for z-gradient correction and higher order xy gradients. PMID- 17411226 TI - Study of thermal parameters' temperature dependence in solids using photothermal radiometry. AB - A photothermal radiometry configuration that allows the measurement of the temperature dependence of thermal parameters of solid materials is described. Two procedures are proposed. The first one is based on a combination of phase and amplitude signal data collected at a single frequency and the second one makes use of the information contained in the phase signal data, obtained at two different chopping frequencies. The methods are recommended for calorimetric studies requiring temperature scans at a constant chopping frequency. PMID- 17411227 TI - Focusing quality of a split short laser pulse. AB - For multiple laser pulse experiments, it is necessary to split a laser pulse. In order to split a short laser pulse without stretching the pulse width, the laser pulse should not pass through thick materials. For this reason, a pellicle beam splitter (BS) and/or a mirror with a hole are required as a BS for the short laser pulse. The focusing qualities of the laser pulse after passing through the pellicle BS and the mirror with a hole are the same as without the BS's. The laser pulse quality reflected by the BSs should be considered for the laser pulse. A pellicle BS is a thin foil, so, it is weak against vibrations. One should be careful about airflows and isolation from vibration sources. The spot size of the reflected laser pulse is consistent with the size reflected by a normal mirror. The energy loss is about 10% compared with a normal mirror. A mirror with a hole is strong against external vibrations. The reflected laser pulse has a doughnut shape. The reflected laser pulse is interfered due to the shape. In order to cleanly focus the laser pulse, the inside size of the doughnut should be smaller than a half size of the outside portion of the doughnut. PMID- 17411228 TI - High-temperature residual stresses in thin films characterized by x-ray diffraction substrate curvature method. AB - A new x-ray technique to determine temperature dependencies of macroscopic stresses in thin films by characterizing the substrate curvature is introduced. The technique is demonstrated on polycrystalline TiN and Al thin films deposited on Si(100) wafers. The structures are thermally cycled in the temperature range of 25-400 degrees C using a newly developed heating chamber attached to a commercial x-ray diffractometer. The curvature of the freestanding samples was determined by the rocking curve measurement of substrate Si 400 reflections at different lateral positions of the samples, and the stresses are calculated using Stoney's formula. The results show that the magnitude of the stress is in good agreement with the results obtained by other techniques. For the practical application of the technique, the sample mounting and the temperature control are of great importance. PMID- 17411229 TI - Rapid rise time pulsed magnetic field circuit for pump-probe field effect studies. AB - Here we describe an electronic circuit capable of producing rapidly switched dc magnetic fields of up to 20 mT with a rise time of 10 ns and a pulse length variable from 50 ns to more than 10 micros, suitable for use in the study of magnetic field effects on radical pair (RP) reactions. This corresponds to switching the field on a time scale short relative to the lifetime of typical RPs and maintaining it well beyond their lifetimes. Previous experiments have involved discharging a capacitor through a low inductance coil for a limited time using a switching circuit. These suffer from decaying field strength over the duration of the pulse given primarily by the ratio of the pulse width to the RC constant of the circuit. We describe here a simple yet elegant solution that completely eliminates this difficulty by employing a feedback loop. This allows a constant field to be maintained over the entire length of the pulse. PMID- 17411230 TI - Magnetic probe array with high sensitivity for fluctuating field. AB - A magnetic probe array is constructed to measure precisely the spatial structure of a small fluctuating field included in a strong confinement field that varies with time. To exclude the effect of the confinement field, the magnetic probes consisting of figure-eight-wound coils are prepared. The spatial structure of the fluctuating field is obtained from a Fourier analysis of the probe signal. It is found that the probe array is more sensitive to the fluctuating field with a high mode number than that with a low mode number. An experimental demonstration of the present method is attempted using a field-reversed configuration plasma, where the fluctuating field with 0.1% of the confinement field is successfully detected. PMID- 17411231 TI - Development of a liquid tin microjet target for an efficient laser-produced plasma extreme ultraviolet source. AB - A regenerative tin liquid microjet target was developed for a high average power extreme ultraviolet (EUV) source. The diameter of the target was smaller than 160 microm and good vacuum lower than 0.5 Pa was maintained during the operation. A maximum EUV conversion efficiency of 1.8% at the Nd:yttrium-aluminum-garnet laser intensity of around 2 x 10(11) Wcm(2) with a spot diameter of 175 microm (full width at half maximum) was observed. The angular distribution of the EUV emission remained almost isotropic, whereas suprathermal ions mainly emerged toward the target normal. PMID- 17411232 TI - Improved parallel scan method for nanofriction force measurement with atomic force microscopy. AB - Based on Ruan and Bhushan's study [J. Ruan and B. Bhushan, J. Tribol. 116, 378 (1994)], an improved method for quantitative nano/microfriction force measurements with the atomic force microscope (AFM) is presented. The related theoretical derivation is given in detail. The coefficient of friction can be calculated by scanning in the direction parallel to the long axis of the AFM cantilever. Then conversion factor, which can convert the lateral deflection response of the photodetector into corresponding friction force, is identified with the Meyer and Amer method [G. Meyer and N. M. Ame, Appl. Phys. Lett. 57, 2089 (1990)]. Like Ruan and Bhushan method, the advantage of this approach is that the coefficient of friction can be obtained with the plan-view geometry of AFM cantilevers and some common uncertainties, such as thickness, coating, and material properties, are not necessary. The result of the experiments performed utilizing rectangular cantilevers of different lengths shows that this improved method produces an accurate agreement for cantilevers of different lengths, thus the method can be used to measure nano/microfriction force. PMID- 17411233 TI - Two-step preionization scheme for discharge pumped KrF excimer lasers. AB - The beam homogeneity and stored energy of discharge pumped KrF excimer amplifiers are critically dependent on the preionization level of the active medium and on the rise time of the electric excitation pulse. Here a scheme is presented to increase the effectiveness of the preionization and to improve the rise time of the pump pulse by the use of the auto-spark-gap UV preionization of increased gap separation. The modified charging circuit has been proven to enhance the discharge quality and the amplification properties of the excimer module. The increased temporal jitter of the preionization sparks of large gap separation is eliminated by an additional pre-preionization scheme. The circuit is especially well suited for short-pulse KrF amplifiers. PMID- 17411234 TI - Precision analog integrator with voltage hold. AB - In some applications, like a magnetization and demagnetization of ordinary and superconducting magnets driven by the voltage controlled current sources, it is desirable to change the voltage smoothly and linearly toward an adjusted value and then hold this value in time. A circuit is presented that allows: (i) the slow, smooth, and linear change of the integrator output voltage without steps and (ii) the ability to hold the output voltage at a value given by a reference source. In a hold regime, the output voltage is stabilized by a negative feedback. A stability of one part of 10(5) at the output voltage of 2 V was achieved over a 12 h period. PMID- 17411235 TI - Vertical inertial sliding drive for coarse and fine approaches in scanning probe microscopy. AB - Mechanisms for controlled approach of a probe tip toward the sample surface are essential in high resolution imaging by scanning probe microscopy (SPM). This work describes the development and performance of an inertial sliding drive capable of translating a relatively large mass (25 g) at up to 1 mms over 1 cm with step sizes of 10-250 nm in ambient conditions using various wave forms as measured by fiber optic interferometry. The drive functions independent of orientation with a threshold voltage of less than 15 V using a single drive signal. Use of piezotube actuators in a radially symmetric arrangement provides guided motion and minimizes differential thermal expansion between critical components. Controlled translation of the entire scanning component in both ambient and electrochemical scanning tunneling microscopy has been routinely achieved with no evidence of tip crash. This device has been specifically designed for use in in situ SPM applications where stability of the sample and that of the liquid environment are paramount. PMID- 17411236 TI - Drift-free atomic force microscopy measurements of cell height and mechanical properties. AB - Atomic force microscopy (AFM) is used to study the morphological and mechanical properties of living cells. However, experiments performed over minutes to hours are subject to significant instrumental drift. The main sources of drift are the cantilever's geometrical asymmetry and bimorphic construction. We developed a simple software Stick-and-Move (SaM) routine for AFM that eliminates drift by continuously referencing the sample position to the substrate while acquiring force-distance curves. Control experiments show no drift over 15 min at an acquisition rate of 0.1 Hz. As a proof of concept, we applied the SaM to study the response of rat astrocytes to osmotic stress, observing dimensional and constitutive changes during volume regulation. PMID- 17411237 TI - The emergence of chaos in a laser irradiated spherical absorber. AB - We show in this paper that the simple system of a spherical absorber immersed in water can exhibit complex and chaotic behavior upon absorption of laser energy. We report on computer experiments performed on this simple system. We present power spectra and calculate Lyapunov exponents that show that for increasing laser pulse durations and increasing laser energy the pressure response of the system changes from periodic to a regime displaying spatiotemporal chaos. This is important from a theoretical point of view because the complex behavior displayed in this simple system makes it an excellent choice for investigations into the nonlinear dynamics of fluids and the complicated transition to turbulence. This is also important for people using these systems for various applications in material science and biomedicine. PMID- 17411238 TI - Nonparaxial effects on the propagation and scattering of a polarized optical pulse. AB - Propagation characteristics of a polarized optical solitary pulse are analyzed by taking into account the effect of nonparaxiality and mutual interaction. To start with, a pair of generalized nonlinear Schrodinger equations is deduced through an operator approach. Stationary solutions of such a system are then analyzed numerically through a boundary value problem in two stages, with and without the nonparaxial effect. In the second stage, the propagating form of the corresponding spatial soliton is studied by an extended split step algorithm ETDRK. The initial profile is considered to be both a one- and two-soliton solution, to visualize the event of scattering and fusion. From this data, we have computed the intensity, root mean square spectral width, and chirp of a single soliton as it propagates. In the case of the two-soliton solution, we observe that for source parameter values, the fusion is more favored than scattering. It is observed that nonparaxiality and the interaction between A(x) and A(y) tends to destroy the periodic behaviors of these parameters. Lastly, we have investigated the modulational instability of the system as function of frequency detuning and nonparaxiality. The form of the gain is discussed as a function of nonparaxiality. PMID- 17411239 TI - Exponential stabilization of chaotic systems with delay by periodically intermittent control. AB - This paper studies the exponential stabilization problem for a class of chaotic systems with delay by means of periodically intermittent control. A unified exponential stability criterion, together with its simplified versions, is established by using Lyapunov function and differential inequality techniques. A suboptimal intermittent controller is designed with respect to the general cost function under the assumption that the control period is fixed. Numerical simulations on two chaotic oscillators are presented to verify the theoretical results. PMID- 17411240 TI - Two-parameter families of strange attractors. AB - Periodically driven two-dimensional nonlinear oscillators can generate strange attractors that are periodic. These attractors are mapped in a locally 1-1 way to entire families of strange attractors that are indexed by a pair of relatively prime integers (n(1),n(2)), with n(1)>/=1. The integers are introduced by imposing periodic boundary conditions on the entire strange attractor rather than individual trajectories in the attractor. The torsion and energy integrals for members of this two parameter family of locally identical strange attractors depend smoothly on these integers. PMID- 17411241 TI - Effect of common noise on phase synchronization in coupled chaotic oscillators. AB - We report a general phenomenon concerning the effect of noise on phase synchronization in coupled chaotic oscillators: the average phase-synchronization time exhibits a nonmonotonic behavior with the noise amplitude. In particular, we find that the time exhibits a local minimum for relatively small noise amplitude but a local maximum for stronger noise. We provide numerical results, experimental evidence from coupled chaotic circuits, and a heuristic argument to establish the generality of this phenomenon. PMID- 17411242 TI - Study on the method of controlling chaos in an Er-doped fiber dual-ring laser via external optical injection and shifting optical feedback light. AB - Controlling chaos in an erbium-doped fiber dual-ring laser is studied by injecting an external light and shifting the phase of an optical feedback light. First, the parameters of the externally injected optical signal are adjusted to bring the chaotic laser into a single-period state and a multiperiod state, respectively. In addition, the single-mode locking in a single ring of the laser as well as the dual-mode locking in dual ring of the laser are found. Secondly, the method of optical feedback is used to stabilize chaos by controlling the parameters of the optical devices in an external optical path. We show that the optical negative feedback can stabilize the chaotic laser into a single-period state, while the optical positive feedback can stabilize the chaotic laser into another single-period state and a period-doubling state. Lastly, various dynamical states are produced in the laser by shifting the phase of the optical feedback light. PMID- 17411243 TI - Numerical computation of orbits and rigorous verification of existence of snapback repellers. AB - In this paper we show how analysis from numerical computation of orbits can be applied to prove the existence of snapback repellers in discrete dynamical systems. That is, we present a computer-assisted method to prove the existence of a snapback repeller of a specific map. The existence of a snapback repeller of a dynamical system implies that it has chaotic behavior [F. R. Marotto, J. Math. Anal. Appl. 63, 199 (1978)]. The method is applied to the logistic map and the discrete predator-prey system. PMID- 17411244 TI - Nonlinear real-life signal detection with a supervised principal components analysis. AB - A novel strategy named supervised principal components analysis for the detection of a target signal of interest embedded in an unknown noisy environment has been investigated. There are two channels in our detection scheme. Each channel consists of a nonlinear phase-space reconstructor (for embedding a data matrix using the received time series) and a principal components analyzer (for feature extraction), respectively. The output error time series, which results from the difference of both eigenvectors of the correlation data matrices from these two channels, is then analyzed using time-frequency tools, for example, frequency spectrum or Wigner-Ville distribution. Experimental results based on real-life electromagnetic data are presented to demonstrate the detection performance of our algorithm. It is found that weak signals hidden beneath the noise floor can be detected. Furthermore, the robustness of the detection performance clearly illustrated that signal frequencies can be extracted when the signal power is not too low. PMID- 17411245 TI - On the classification of buoyancy-driven chemo-hydrodynamic instabilities of chemical fronts. AB - Exothermic autocatalytic fronts traveling in the gravity field can be deformed by buoyancy-driven convection due to solutal and thermal contributions to changes in the density of the product versus the reactant solutions. We classify the possible instability mechanisms, such as Rayleigh-Benard, Rayleigh-Taylor, and double-diffusive mechanisms known to operate in such conditions in a parameter space spanned by the corresponding solutal and thermal Rayleigh numbers. We also discuss a counterintuitive instability leading to buoyancy-driven deformation of statically stable fronts across which a solute-light and hot solution lies on top of a solute-heavy and colder one. The mechanism of this chemically driven instability lies in the coupling of a localized reaction zone and of differential diffusion of heat and mass. Dispersion curves of the various cases are analyzed. A discussion of the possible candidates of autocatalytic reactions and experimental conditions necessary to observe the various instability scenarios is presented. PMID- 17411246 TI - A unified approach to attractor reconstruction. AB - In the analysis of complex, nonlinear time series, scientists in a variety of disciplines have relied on a time delayed embedding of their data, i.e., attractor reconstruction. The process has focused primarily on intuitive, heuristic, and empirical arguments for selection of the key embedding parameters, delay and embedding dimension. This approach has left several longstanding, but common problems unresolved in which the standard approaches produce inferior results or give no guidance at all. We view the current reconstruction process as unnecessarily broken into separate problems. We propose an alternative approach that views the problem of choosing all embedding parameters as being one and the same problem addressable using a single statistical test formulated directly from the reconstruction theorems. This allows for varying time delays appropriate to the data and simultaneously helps decide on embedding dimension. A second new statistic, undersampling, acts as a check against overly long time delays and overly large embedding dimension. Our approach is more flexible than those currently used, but is more directly connected with the mathematical requirements of embedding. In addition, the statistics developed guide the user by allowing optimization and warning when embedding parameters are chosen beyond what the data can support. We demonstrate our approach on uni- and multivariate data, data possessing multiple time scales, and chaotic data. This unified approach resolves all the main issues in attractor reconstruction. PMID- 17411247 TI - Revealing direction of coupling between neuronal oscillators from time series: phase dynamics modeling versus partial directed coherence. AB - The problem of determining directional coupling between neuronal oscillators from their time series is addressed. We compare performance of the two well established approaches: partial directed coherence and phase dynamics modeling. They represent linear and nonlinear time series analysis techniques, respectively. In numerical experiments, we found each of them to be applicable and superior under appropriate conditions: The latter technique is superior if the observed behavior is "closer" to limit-cycle dynamics, the former is better in cases that are closer to linear stochastic processes. PMID- 17411248 TI - Delay time modulation induced oscillating synchronization and intermittent anticipatory/lag and complete synchronizations in time-delay nonlinear dynamical systems. AB - Existence of a new type of oscillating synchronization that oscillates between three different types of synchronizations (anticipatory, complete, and lag synchronizations) is identified in unidirectionally coupled nonlinear time-delay systems having two different time-delays, that is feedback delay with a periodic delay time modulation and a constant coupling delay. Intermittent anticipatory, intermittent lag, and complete synchronizations are shown to exist in the same system with identical delay time modulations in both the delays. The transition from anticipatory to complete synchronization and from complete to lag synchronization as a function of coupling delay with suitable stability condition is discussed. The intermittent anticipatory and lag synchronizations are characterized by the minimum of the similarity functions and the intermittent behavior is characterized by a universal asymptotic -32 power law distribution. It is also shown that the delay time carved out of the trajectories of the time delay system with periodic delay time modulation cannot be estimated using conventional methods, thereby reducing the possibility of decoding the message by phase space reconstruction. PMID- 17411249 TI - Dynamics of chaotic driving: rotation in the restricted three-body problem. AB - We investigate the rotation of a small nonspherical body in the planar restricted three-body problem along periodic, quasi-periodic, and chaotic orbits of the small body's center of mass. The rotation dynamics is chaotic in all three cases, but a systematic overview of it via stroboscopic mappings is possible only in the periodic case. We propose to explore the structured phase space patterns by following an ensemble of trajectories, a droplet, in the phase space. The temporal evolution of the pattern can be characterized by a time-dependent fractal dimension. It is shown to converge exponentially to a time-independent value for long times. In the presence of dissipation, the droplet typically converges to a so-called snapshot chaotic attractor whose shape might change chaotically in time, but whose asymptotic fractal dimension is constant. PMID- 17411250 TI - Dispersive destabilization of nonlinear light propagation in fiber Bragg gratings: a numerical verification. AB - This paper presents some numerical simulations of the full one-dimensional Maxwell-Lorentz equations that describe light propagation in fiber Bragg gratings in order to confirm that the standard nonlinear coupled mode equations fail to predict the weakly nonlinear dynamics of the system when dispersive instabilities come into play, and that, in this case, the correct slow envelope description of the system requires us to consider higher order dispersion effects. PMID- 17411251 TI - Recurrence plots and Shannon entropy for a dynamical analysis of asynchronisms in noninvasive mechanical ventilation. AB - Recurrence plots were introduced to quantify the recurrence properties of chaotic dynamics. Hereafter, the recurrence quantification analysis was introduced to transform graphical interpretations into statistical analysis. In this spirit, a new definition for the Shannon entropy was recently introduced in order to have a measure correlated with the largest Lyapunov exponent. Recurrence plots and this Shannon entropy are thus used for the analysis of the dynamics underlying patient assisted with a mechanical noninvasive ventilation. The quality of the assistance strongly depends on the quality of the interactions between the patient and his ventilator which are crucial for tolerance and acceptability. Recurrence plots provide a global view of these interactions and the Shannon entropy is shown to be a measure of the rate of asynchronisms as well as the breathing rhythm. PMID- 17411252 TI - "Metric" complexity for weakly chaotic systems. AB - We consider the number of Bowen sets necessary to cover a large measure subset of the phase space. This introduces some complexity indicator characterizing different kinds of (weakly) chaotic dynamics. Since in many systems its value is given by a sort of local entropy, this indicator is quite simple to calculate. We give some examples of calculations in nontrivial systems (e.g., interval exchanges and piecewise isometries) and a formula similar to that of Ruelle Pesin, relating the complexity indicator to some initial condition sensitivity indicators playing the role of positive Lyapunov exponents. PMID- 17411253 TI - Anticipating synchronization of chaotic Lur'e systems. AB - In this paper we consider the anticipating synchronization of chaotic time delayed Lur'e-type systems in a master-slave setting. We introduce three scenarios for anticipating synchronization, and give sufficient conditions for the existence of anticipating synchronizing slave systems in terms of linear matrix inequalities. The results obtained are illustrated on a time-delayed Rossler system and a time-delayed Chua oscillator. PMID- 17411254 TI - Multifolded torus chaotic attractors: design and implementation. AB - This paper proposes a systematic methodology for creating multifolded torus chaotic attractors from a simple three-dimensional piecewise-linear system. Theoretical analysis shows that the multifolded torus chaotic attractors can be generated via alternative switchings between two basic linear systems. The theoretical design principle and the underlying dynamic mechanism are then further investigated by analyzing the emerging bifurcation and the stable and unstable subspaces of the two basic linear systems. A novel block circuit diagram is also designed for hardware implementation of 3-, 5-, 7-, 9-folded torus chaotic attractors via switching the corresponding switches. This is the first time a 9-folded torus chaotic attractor generated by an analog circuit has been verified experimentally. Furthermore, some recursive formulas of system parameters are rigorously derived, which is useful for improving hardware implementation. PMID- 17411255 TI - Dynamical properties of a dissipative hybrid Fermi-Ulam-bouncer model. AB - Some consequences of dissipation are studied for a classical particle suffering inelastic collisions in the hybrid Fermi-Ulam bouncer model. The dynamics of the model is described by a two-dimensional nonlinear area-contracting map. In the limit of weak and moderate dissipation we report the occurrence of crisis and in the limit of high dissipation the model presents doubling bifurcation cascades. Moreover, we show a phenomena of annihilation by pairs of fixed points as the dissipation varies. PMID- 17411256 TI - Bifurcation analysis of bubble dynamics in fluidized beds. AB - We use a low-dimensional, agent-based bubble model to study the changes in the global dynamics of fluidized beds in response to changes in the frequency of the rising bubbles. The computationally based bifurcation analysis shows that at low frequencies, the global dynamics is attracted towards a fixed point since the bubbles interact very little with one another. As the frequency of injection increases, however, the global dynamics undergoes a series of bifurcations to new behaviors that include highly periodic orbits, chaotic attractors, and intermittent behavior between periodic orbits and chaotic sets. Using methods from time-series analysis, we are able to approximate nonlinear models that allow for long-term predictions and the possibility of developing control algorithms. PMID- 17411257 TI - Art of war hidden in Kolmogorov's equations. AB - Here we discuss how Kolmogorov's work on turbulence can be used as the inspiration for a new description of battlefield dynamics. The method presented may also represent a new way of describing self-organizing dynamical systems, in place of conventional differential equation approaches. The key finding is that the rate of attrition in a battle appears to be a function of the fractal dimension of the opposing forces. It is suggested that, this being the case, the fractal dimension could be used as a surrogate to represent the organizational efficiency of one force relative to another, commonly called Command and Control. PMID- 17411258 TI - Introduction: Cardiovascular physics. AB - The number of patients suffering from cardiovascular diseases increases unproportionally high with the increase of the human population and aging, leading to very high expenses in the public health system. Therefore, the challenge of cardiovascular physics is to develop high-sophisticated methods which are able to, on the one hand, supplement and replace expensive medical devices and, on the other hand, improve the medical diagnostics with decreasing the patient's risk. Cardiovascular physics-which interconnects medicine, physics, biology, engineering, and mathematics-is based on interdisciplinary collaboration of specialists from the above scientific fields and attempts to gain deeper insights into pathophysiology and treatment options. This paper summarizes advances in cardiovascular physics with emphasis on a workshop held in Bad Honnef, Germany, in May 2005. The meeting attracted an interdisciplinary audience and led to a number of papers covering the main research fields of cardiovascular physics, including data analysis, modeling, and medical application. The variety of problems addressed by this issue underlines the complexity of the cardiovascular system. It could be demonstrated in this Focus Issue, that data analyses and modeling methods from cardiovascular physics have the ability to lead to significant improvements in different medical fields. Consequently, this Focus Issue of Chaos is a status report that may invite all interested readers to join the community and find competent discussion and cooperation partners. PMID- 17411259 TI - Negative filament tension in the Luo-Rudy model of cardiac tissue. AB - Scroll waves are vortices that occur in three-dimensional excitable media. Scroll waves have been observed in a variety of systems including cardiac tissue, where they are associated with cardiac arrhythmias. The disorganization of scroll waves into chaotic behavior is thought to be the mechanism of ventricular fibrillation, which lethality is widely known. One of the possible mechanisms of scroll wave instability is negative filament tension, which was studied theoretically using low-dimensional models of excitable medium. In this article we perform a numerical study of negative filament tension using the Luo-Rudy phase 1 model, which is widely used in cardiac electrophysiology. We show that this instability exists in this model, study its manifestation and discuss its relation to cardiac arrhythmogenesis. PMID- 17411260 TI - Arrhythmogenesis in the heart: Multiscale modeling of the effects of defibrillation shocks and the role of electrophysiological heterogeneity. AB - The mechanisms of initiation of ventricular arrhythmias as well as those behind the complex spatiotemporal wave dynamics and its filament organization during ventricular fibrillation (VF) are the topic of intense research and debate. Mechanistic inquiry into the various mechanisms that lead to arrhythmia initiation and VF maintenance is hampered by the inability of current experimental techniques to resolve, with sufficient accuracy, electrical behavior confined to the depth of the ventricles. The objective of this article is to demonstrate that realistic 3D simulations of electrical activity in the heart are capable of bringing a new level of understanding of the mechanisms that underlie arrhythmia initiation and subsequent organization. The article does this by presenting the results of two multiscale simulation studies of ventricular electrical behavior. The first study aims to uncover the mechanisms responsible for rendering the ventricles vulnerable to electric shocks during a specific interval of time, the vulnerable window. The second study focuses on elucidating the role of electrophysiological heterogeneity, and specifically, differences in action potential duration in various ventricular structures, in VF organization. Both studies share common multiscale modeling approaches and analysis, including characterization of scroll-wave filament dynamics. PMID- 17411261 TI - Alternans and the influence of ionic channel modifications: Cardiac three dimensional simulations and one-dimensional numerical bifurcation analysis. AB - Cardiac propagation is investigated by simulations using a realistic three dimensional (3D) geometry including muscle fiber orientation of the ventricles of a rabbit heart and the modified Beeler-Reuter ionic model. Electrical excitation is introduced by a periodic pacing of the lower septum. Depending on the pacing frequency, qualitatively different dynamics are observed, namely, normal heart beat, T-wave alternans, and 2:1 conduction block at small, intermediate, and large pacing frequencies, respectively. In a second step, we performed a numerical stability and bifurcation analysis of a pulse propagating in a one dimensional (1D) ring of cardiac tissue. The precise onset of the alternans instability is obtained from computer-assisted linear stability analysis of the pulse and computation of the associated spectrum. The critical frequency at the onset of alternans and the profiles of the membrane potential agree well with the ones obtained in the 3D simulations. Next, we computed changes in the wave profiles and in the onset of alternans for the Beeler-Reuter model with modifications of the sodium, calcium, and potassium channels, respectively. For this purpose, we employ the method of numerical bifurcation and stability analysis. While blocking of calcium channels has a stabilizing effect, blocked sodium or potassium channels lead to the occurrence of alternans at lower pacing frequencies. The findings regarding channel blocking are verified within three dimensional simulations. Altogether, we have found T-wave alternans and conduction block in 3D simulations of a realistic rabbit heart geometry. The onset of alternans has been analyzed by numerical bifurcation and stability analysis of 1D wave trains. By comparing the results of the two approaches, we find that alternans is not strongly influenced by ingredients such as 3D geometry and propagation anisotropy, but depends mostly on the frequency of pacing (frequency of subsequent action potentials). In addition, we have introduced numerical bifurcation and stability analysis as a tool into heart modeling and demonstrated its efficiency in scanning a large set of parameters in the case of models with reduced conductivity. Bifurcation analysis also provides an accurate test for analytical theories of alternans as is demonstrated for the case of the restitution hypothesis. PMID- 17411262 TI - Virtual cell and tissue dynamics of ectopic activation of the ventricles. AB - Cardiac ventricular cells and tissues are normally excitable, and are activated by propagating waves of excitation that are initiated in the specialized pacemaking region of the heart. However, isolated or repetitive activity can be initiated at abnormal (ectopic) sites in the ventricles. To trigger an endogenous ectopic beat, there must be a compact focus of cells with changed membrane excitation parameters and kinetics, which initiate activity by after depolarizations triggered by propagating activity, or that have bifurcated into autorhythmicity. This ectopic focus needs to be large enough, and adequately coupled, to drive the surrounding tissue. We investigate the initiation of ectopic excitation in computational models of human ventricular cells triggered by after-depolarizations and by up/down regulation of specific membrane conductance systems, and the propagation and evolution of ectopic activity in homogeneous or heterogeneous and isotropic, anisotropic, or orthotropic tissues. PMID- 17411263 TI - Cardiac resynchronization therapy and phase resetting of the sinoatrial node: a conjecture. AB - Congestive heart failure is a severe chronic disease often associated with disorders that alter the mechanisms of excitation-contraction coupling that may result in an asynchronous left ventricular motion which may further impair the ability of the failing heart to eject blood. In recent years a therapeutic approach to resynchronize the ventricles (cardiac resynchronization therapy, CRT) has been performed through the use of a pacemaker device able to provide atrial based biventricular stimulation. Atrial lead senses the spontaneous occurrence of cells depolarization and sends the information to the generator which, in turn, after a settled delay [atrioventricular (AV) delay], sends electrical impulses to both ventricles to stimulate their synchronous contraction. Recent studies performed on heart rate behavior of chronically implanted patients at different epochs after implantation have shown that CRT can lead to sustained overall improvement of heart function with a reduction in morbidity and mortality. At this moment, however, there are no studies about CRT effects on spontaneous heart activity of chronically implanted patients. We performed an experimental study in which the electrocardiographic signal of five subjects under chronic CRT was recorded during the activity of the pacemaker programmed at different AV delays and under spontaneous cardiac activity after pacemaker deactivation. The different behavior of heart rate variability during pacemaker activity and after pacemaker deactivation suggested the hypothesis of a phase resetting mechanism induced by the pacemaker stimulus on the sinoatrial (SA) node, a phenomenon already known in literature for aggregate of cardiac cells, but still unexplored in vivo. The constraints imposed by the nature of our study (in vivo tests) made it impossible to plan an experiment to prove our hypothesis directly. We therefore considered the best attainable result would be to prove the accordance of our data to the conjecture through the use of models and physical considerations. We first used the data of literature on far-field effects of cardiac defibrillators to prove that the pacemaker impulses delivered to the two ventricles were able to induce modifications in membrane voltage at the level of the SA node. To simulate a phase resetting mechanism of the SA node, we used a Van der Pol modified model to allow the possibility of changing the refractory period and the firing frequency of the cells separately. With appropriate parameters of the model we reproduced phase response curves that can account for our experimental data. Furthermore, the simulated curves closely resemble the functional form proposed in literature for perturbed aggregate of cardiac cells. Despite the small sample of subjects investigated and the limited number of ECG recordings at different AV delays, we think we have proved the plausibility of the proposed conjecture. PMID- 17411264 TI - Turbulence control with local pacing and its implication in cardiac defibrillation. AB - In this review article, we describe turbulence control in excitable systems by using a local periodic pacing method. The controllability conditions of turbulence suppression and the mechanisms underlying these conditions are analyzed. The local pacing method is applied to control Winfree turbulence (WT) and defect turbulence (DT) induced by spiral-wave breakup. It is shown that WT can always be suppressed by local pacing if the pacing amplitude and frequency are properly chosen. On the other hand, the pacing method can achieve suppression of DT induced by instabilities associated with the motions of spiral tips while failing to suppress DT induced by the instabilities of wave propagation far from tips. In the latter case, an auxiliary method of applying gradient field is suggested to improve the control effects. The implication of this local pacing method to realistic cardiac defibrillation is addressed. PMID- 17411265 TI - Long-term invariant parameters obtained from 24-h Holter recordings: a comparison between different analysis techniques. AB - Over the last two decades, a large number of different methods had been used to study the fractal-like behavior of the heart rate variability (HRV). In this paper some of the most used techniques were reviewed. In particular, the focus is set on those methods which characterize the long memory behavior of time series (in particular, periodogram, detrended fluctuation analysis, rescale range analysis, scaled window variance, Higuchi dimension, wavelet-transform modulus maxima, and generalized structure functions). The performances of the different techniques were tested on simulated self-similar noises (fBm and fGn) for values of alpha, the slope of the spectral density for very small frequency, ranging from -1 to 3 with a 0.05 step. The check was performed using the scaling relationships between the various indices. DFA and periodogram showed the smallest mean square error from the expected values in the range of interest for HRV. Building on the results obtained from these tests, the effective ability of the different methods in discriminating different populations of patients from RR series derived from Holter recordings, was assessed. To this extent, the Noltisalis database was used. It consists of a set of 30, 24-h Holter recordings collected from healthy subjects, patients suffering from congestive heart failure, and heart transplanted patients. All the methods, with the exception at most of rescale range analysis, were almost equivalent in distinguish between the three groups of patients. Finally, the scaling relationships, valid for fBm and fGn, when empirically used on HRV series, also approximately held. PMID- 17411266 TI - Patterns of spiral wave attenuation by low-frequency periodic planar fronts. AB - There is evidence that spiral waves and their breakup underlie mechanisms related to a wide spectrum of phenomena ranging from spatially extended chemical reactions to fatal cardiac arrhythmias [A. T. Winfree, The Geometry of Biological Time (Springer-Verlag, New York, 2001); J. Schutze, O. Steinbock, and S. C. Muller, Nature 356, 45 (1992); S. Sawai, P. A. Thomason, and E. C. Cox, Nature 433, 323 (2005); L. Glass and M. C. Mackey, From Clocks to Chaos: The Rhythms of Life (Princeton University Press, Princeton, 1988); R. A. Gray et al., Science 270, 1222 (1995); F. X. Witkowski et al., Nature 392, 78 (1998)]. Once initiated, spiral waves cannot be suppressed by periodic planar fronts, since the domains of the spiral waves grow at the expense of the fronts [A. N. Zaikin and A. M. Zhabotinsky, Nature 225, 535 (1970); A. T. Stamp, G. V. Osipov, and J. J. Collins, Chaos 12, 931 (2002); I. Aranson, H. Levine, and L. Tsimring, Phys. Rev. Lett. 76, 1170 (1996); K. J. Lee, Phys. Rev. Lett. 79, 2907 (1997); F. Xie, Z. Qu, J. N. Weiss, and A. Garfinkel, Phys. Rev. E 59, 2203 (1999)]. Here, we show that introducing periodic planar waves with long excitation duration and a period longer than the rotational period of the spiral can lead to spiral attenuation. The attenuation is not due to spiral drift and occurs periodically over cycles of several fronts, forming a variety of complex spatiotemporal patterns, which fall into two distinct general classes. Further, we find that these attenuation patterns only occur at specific phases of the descending fronts relative to the rotational phase of the spiral. We demonstrate these dynamics of phase-dependent spiral attenuation by performing numerical simulations of wave propagation in the excitable medium of myocardial cells. The effect of phase-dependent spiral attenuation we observe can lead to a general approach to spiral control in physical and biological systems with relevance for medical applications. PMID- 17411267 TI - Interactions between short-term and long-term cardiovascular control mechanisms. AB - The cardiovascular system incorporates several controlling mechanisms acting as feedback loops over different time horizons. Because of their complex interrelationships, information-based methods such as autonomic information flow (AIF) functions promise to be useful in identifying normal and pathological behavior. Optimal adjustment between those controllers is necessary for healthy global behavior of the organism. We investigated the question as to whether there are typical relationships between short-term and long-term AIF by means of a meta analysis of several of our own clinical studies of the mortality of patients with multiple organ dysfunction syndrome, heart failure, idiopathic dilated cardiomyopathy, and the length of stay in hospital after abdominal aorta surgery. We found a fundamental association of increased short-term randomness (decreased AIF) and decreased long-term randomness (increased AIF) due to pathology. A systems theoretic validation of this fundamental type of association was done by an appropriate mathematical model using a dissipative system with two feedback loops over different time horizons. The systematic simulation of an increasing collapse of the short feedback loop confirmed the inverse association between short-term and long-term information flow as a fundamental, system inherent type of readjustment that occurs under pathological conditions. PMID- 17411268 TI - Cluster synchronization and spatio-temporal dynamics in networks of oscillatory and excitable Luo-Rudy cells. AB - We study collective phenomena in nonhomogeneous cardiac cell culture models, including one- and two-dimensional lattices of oscillatory cells and mixtures of oscillatory and excitable cells. Individual cell dynamics is described by a modified Luo-Rudy model with depolarizing current. We focus on the transition from incoherent behavior to global synchronization via cluster synchronization regimes as coupling strength is increased. These regimes are characterized qualitatively by space-time plots and quantitatively by profiles of local frequencies and distributions of cluster sizes in dependence upon coupling strength. We describe spatio-temporal patterns arising during this transition, including pacemakers, spiral waves, and complicated irregular activity. PMID- 17411269 TI - Phase-rectified signal averaging for the detection of quasi-periodicities and the prediction of cardiovascular risk. AB - We present the phase-rectified signal averaging (PRSA) method as an efficient technique for the study of quasi-periodic oscillations in noisy, nonstationary signals. It allows the assessment of system dynamics despite phase resetting and noise and in relation with either increases or decreases of the considered signal. We employ the method to study the quasi-periodicities of the human heart rate based on long-term ECG recordings. The center deflection of the PRSA curve characterizes the average capacity of the heart to decelerate (or accelerate) the cardiac rhythm. It can be measured by a central wavelet coefficient which we denote as deceleration capacity (DC). We find that decreased DC is a more precise predictor of mortality in survivors of heart attack than left ventricular ejection fraction, the current "gold standard" risk predictor. In addition, we discuss the dependence of the DC parameter on age and on diabetes. PMID- 17411270 TI - Analysis of cardiovascular oscillations: a new approach to the early prediction of pre-eclampsia. AB - Pre-eclampsia (PE) is a serious disorder with high morbidity and mortality occurring during pregnancy; 3%-5% of all pregnant women are affected. Early prediction is still insufficient in clinical practice. Although most pre eclamptic patients show pathological uterine perfusion in the second trimester, this parameter has a positive predictive accuracy of only 30%, which makes it unsuitable for early, reliable prediction. The study is based on the hypothesis that alterations in cardiovascular regulatory behavior can be used to predict PE. Ninety-six pregnant women in whom Doppler investigation detected perfusion disorders of the uterine arteries were included in the study. Twenty-four of these pregnant women developed PE after the 30th week of gestation. During pregnancy, additional several noninvasive continuous blood pressure recordings were made over 30 min under resting conditions by means of a finger cuff. The time series extracted of systolic as well as diastolic beat-to-beat pressures and the heart rate were studied by variability and coupling analysis to find predictive factors preceding genesis of the disease. In the period between the 18th and 26th weeks of pregnancy, three special variability and baroreflex parameters were able to predict PE several weeks before clinical manifestation. Discriminant function analysis of these parameters was able to predict PE with a sensitivity and specificity of 87.5% and a positive predictive value of 70%. The combined clinical assessment of uterine perfusion and cardiovascular variability demonstrates the best current prediction several weeks before clinical manifestation of PE. PMID- 17411271 TI - Vascular coupling induces synchronization, quasiperiodicity, and chaos in a nephron tree. AB - The paper presents a study of synchronization phenomena in a system of 22 nephrons supplied with blood from a common cortical radial artery. The nephrons are assumed to interact via hemodynamic and vascularly propagated coupling, both mediated by vascular connections. Using anatomic and physiological criteria, the nephrons are divided into groups: cortical nephrons and medullary nephrons with short, intermediate and long Henle loops. Within each of these groups the delay parameters of the internal feedback regulation are given a random component to represent the internephron variability. For parameters that generate simple limit cycle dynamics in the pressure and flow regulation of single nephrons, the ensemble of coupled nephrons showed steady state, quasiperiodic or chaotic dynamics, depending on the interaction strengths and the arterial blood pressure. When the solutions were either quasiperiodic or chaotic, cortical nephrons synchronized to a single frequency, but the longer medullary nephrons formed two clusters with different frequencies. Under no physiologically realistic combination of parameters did all nephrons assume a common frequency. Our results suggest a greater variability in the nephron dynamics than is apparent from measurements performed on cortical nephrons only. This variability may explain the development of chaotic dynamics in tubular pressure records from hypertensive rats. PMID- 17411272 TI - Statistical physics approach to categorize biologic signals: from heart rate dynamics to DNA sequences. AB - We recently proposed a novel approach to categorize information carried by symbolic sequences based on their usage of repetitive patterns. A simple quantitative index to measure the dissimilarity between two symbolic sequences can be defined. This information dissimilarity index, defined by our formula, is closely related to the Shannon entropy and rank order of the repetitive patterns in the symbolic sequences. Here we discuss the underlying statistical physics assumptions of this dissimilarity index. We use human cardiac interbeat interval time series and DNA sequences as examples to illustrate the applicability of this generic approach to real-world problems. PMID- 17411273 TI - Cardiovascular and respiratory dynamics during normal and pathological sleep. AB - Sleep is an active and regulated process with restorative functions for physical and mental conditions. Based on recordings of brain waves and the analysis of characteristic patterns and waveforms it is possible to distinguish wakefulness and five sleep stages. Sleep and the sleep stages modulate autonomous nervous system functions such as body temperature, respiration, blood pressure, and heart rate. These functions consist of a sympathetic tone usually related to activation and to parasympathetic (or vagal) tone usually related to inhibition. Methods of statistical physics are used to analyze heart rate and respiration to detect changes of the autonomous nervous system during sleep. Detrended fluctuation analysis and synchronization analysis and their applications to heart rate and respiration during sleep in healthy subjects and patients with sleep disorders are presented. The observed changes can be used to distinguish sleep stages in healthy subjects as well as to differentiate normal and disturbed sleep on the basis of heart rate and respiration recordings without direct recording of brain waves. Of special interest are the cardiovascular consequences of disturbed sleep because they present a risk factor for cardiovascular disorders such as arterial hypertension, cardiac ischemia, sudden cardiac death, and stroke. New derived variables can help to find indicators for these health risks. PMID- 17411274 TI - An integrated approach based on uniform quantization for the evaluation of complexity of short-term heart period variability: Application to 24 h Holter recordings in healthy and heart failure humans. AB - We propose an integrated approach based on uniform quantization over a small number of levels for the evaluation and characterization of complexity of a process. This approach integrates information-domain analysis based on entropy rate, local nonlinear prediction, and pattern classification based on symbolic analysis. Normalized and non-normalized indexes quantifying complexity over short data sequences ( approximately 300 samples) are derived. This approach provides a rule for deciding the optimal length of the patterns that may be worth considering and some suggestions about possible strategies to group patterns into a smaller number of families. The approach is applied to 24 h Holter recordings of heart period variability derived from 12 normal (NO) subjects and 13 heart failure (HF) patients. We found that: (i) in NO subjects the normalized indexes suggest a larger complexity during the nighttime than during the daytime; (ii) this difference may be lost if non-normalized indexes are utilized; (iii) the circadian pattern in the normalized indexes is lost in HF patients; (iv) in HF patients the loss of the day-night variation in the normalized indexes is related to a tendency of complexity to increase during the daytime and to decrease during the nighttime; (v) the most likely length L of the most informative patterns ranges from 2 to 4; (vi) in NO subjects classification of patterns with L=3 indicates that stable patterns (i.e., those with no variations) are more present during the daytime, while highly variable patterns (i.e., those with two unlike variations) are more frequent during the nighttime; (vii) during the daytime in HF patients, the percentage of highly variable patterns increases with respect to NO subjects, while during the nighttime, the percentage of patterns with one or two like variations decreases. PMID- 17411275 TI - Cardiac magnetic field map topology quantified by Kullback-Leibler entropy identifies patients with hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is a common primary inherited cardiac muscle disorder, defined clinically by the presence of unexplained left ventricular hypertrophy. The detection of affected patients remains challenging. Genetic testing is limited because only in 50%-60% of all HCM diagnoses an underlying mutation can be found. Furthermore, the disease has a varied clinical course and outcome, with many patients having little or no discernible cardiovascular symptoms, whereas others develop profound exercise limitation and recurrent arrhythmias or sudden cardiac death. Therefore prospective screening of HCM family members is strongly recommended. According to the current guidelines this includes serial echocardiographic and electrocardiographic examinations. In this study we investigated the capability of cardiac magnetic field mapping (CMFM) to detect patients suffering from HCM. We introduce for the first time a combined diagnostic approach based on map topology quantification using Kullback-Leibler (KL) entropy and regional magnetic field strength parameters. The cardiac magnetic field was recorded over the anterior chest wall using a multichannel-LT SQUID system. CMFM was calculated based on a regular 36 point grid. We analyzed CMFM in patients with confirmed diagnosis of HCM (HCM, n=33, 43.8+/-13 years, 13 women, 20 men), a control group of healthy subjects (NORMAL, n=57, 39.6+/-8.9 years; 22 women and 35 men), and patients with confirmed cardiac hypertrophy due to arterial hypertension (HYP, n=42, 49.7+/-7.9 years, 15 women and 27 men). A subgroup analysis was performed between HCM patients suffering from the obstructive (HOCM, n=19) and nonobstructive (HNCM, n=14) form of the disease. KL entropy based map topology quantification alone identified HCM patients with a sensitivity of 78.8% and specificity of 86.9% (overall classification rate 84.8%). The combination of the KL parameters with a regional field strength parameter improved the overall classification rate to 87.9% (sensitivity: 84.8%, specificity: 88.9%, area under ROC curve: 0.94). KL measures applied to discriminate between HOCM and HNCM patients showed a correct classification of 78.8%. The combination of one KL and one regional parameter again improved the overall classification rate to 97%. A preliminary prospective analysis in two HCM families showed the feasibility of this diagnostic approach with a correct diagnosis of all 22 screened family members (1 HOCM, 4 HNCM, 17 normal). In conclusion, Cardiac Magnetic Field Mapping including KL entropy based topology quantifications is a suitable tool for HCM screening. PMID- 17411276 TI - Quantification of fetal heart rate regularity using symbolic dynamics. AB - Fetal heart rate complexity was examined on the basis of RR interval time series obtained in the second and third trimester of pregnancy. In each fetal RR interval time series, short term beat-to-beat heart rate changes were coded in 8 bit binary sequences. Redundancies of the 2(8) different binary patterns were reduced by two different procedures. The complexity of these sequences was quantified using the approximate entropy (ApEn), resulting in discrete ApEn values which were used for classifying the sequences into 17 pattern sets. Also, the sequences were grouped into 20 pattern classes with respect to identity after rotation or inversion of the binary value. There was a specific, nonuniform distribution of the sequences in the pattern sets and this differed from the distribution found in surrogate data. In the course of gestation, the number of sequences increased in seven pattern sets, decreased in four and remained unchanged in six. Sequences that occurred less often over time, both regular and irregular, were characterized by patterns reflecting frequent beat-to-beat reversals in heart rate. They were also predominant in the surrogate data, suggesting that these patterns are associated with stochastic heart beat trains. Sequences that occurred more frequently over time were relatively rare in the surrogate data. Some of these sequences had a high degree of regularity and corresponded to prolonged heart rate accelerations or decelerations which may be associated with directed fetal activity or movement or baroreflex activity. Application of the pattern classes revealed that those sequences with a high degree of irregularity correspond to heart rate patterns resulting from complex physiological activity such as fetal breathing movements. The results suggest that the development of the autonomic nervous system and the emergence of fetal behavioral states lead to increases in not only irregular but also regular heart rate patterns. Using symbolic dynamics to examine the cardiovascular system may thus lead to new insight with respect to fetal development. PMID- 17411277 TI - Comparison of nonlinear methods symbolic dynamics, detrended fluctuation, and Poincare plot analysis in risk stratification in patients with dilated cardiomyopathy. AB - Dilated cardiomyopathy (DCM) has an incidence of about 20100 000 new cases per annum and accounts for nearly 10 000 deaths per year in the United States. Approximately 36% of patients with dilated cardiomyopathy (DCM) suffer from cardiac death within five years after diagnosis. Currently applied methods for an early risk prediction in DCM patients are rather insufficient. The objective of this study was to investigate the suitability of short-term nonlinear methods symbolic dynamics (STSD), detrended fluctuation (DFA), and Poincare plot analysis (PPA) for risk stratification in these patients. From 91 DCM patients and 30 healthy subjects (REF), heart rate and blood pressure variability (HRV, BPV), STSD, DFA, and PPA were analyzed. Measures from BPV analysis, DFA, and PPA revealed highly significant differences (p<0.0011) discriminating REF and DCM. For risk stratification in DCM patients, four parameters from BPV analysis, STSD, and PPA revealed significant differences between low and high risk (maximum sensitivity: 90%, specificity: 90%). These results suggest that STSD and PPA are useful nonlinear methods for enhanced risk stratification in DCM patients. PMID- 17411278 TI - Nonlinear oscillator model reproducing various phenomena in the dynamics of the conduction system of the heart. AB - A dedicated nonlinear oscillator model able to reproduce the pulse shape, refractory time, and phase sensitivity of the action potential of a natural pacemaker of the heart is developed. The phase space of the oscillator contains a stable node, a hyperbolic saddle, and an unstable focus. The model reproduces several phenomena well known in cardiology, such as certain properties of the sinus rhythm and heart block. In particular, the model reproduces the decrease of heart rate variability with an increase in sympathetic activity. A sinus pause occurs in the model due to a single, well-timed, external pulse just as it occurs in the heart, for example due to a single supraventricular ectopy. Several ways by which the oscillations cease in the system are obtained (models of the asystole). The model simulates properly the way vagal activity modulates the heart rate and reproduces the vagal paradox. Two such oscillators, coupled unidirectionally and asymmetrically, allow us to reproduce the properties of heart rate variability obtained from patients with different kinds of heart block including sino-atrial blocks of different degree and a complete AV block (third degree). Finally, we demonstrate the possibility of introducing into the model a spatial dimension that creates exciting possibilities of simulating in the future the SA the AV nodes and the atrium including their true anatomical structure. PMID- 17411300 TI - Mitochondria: more than mitochondrial DNA in cancer. PMID- 17411301 TI - Clinical trials registration. PMID- 17411312 TI - Worldmapper: the human anatomy of a small planet. PMID- 17411313 TI - Noncommunicable diseases and injuries: action needed in South Asia too. PMID- 17411314 TI - The relationship of previous training and experience of journal peer reviewers to subsequent review quality. AB - BACKGROUND: Peer review is considered crucial to the selection and publication of quality science, but very little is known about the previous experiences and training that might identify high-quality peer reviewers. The reviewer selection processes of most journals, and thus the qualifications of their reviewers, are ill defined. More objective selection of peer reviewers might improve the journal peer review process and thus the quality of published science. METHODS AND FINDINGS: 306 experienced reviewers (71% of all those associated with a specialty journal) completed a survey of past training and experiences postulated to improve peer review skills. Reviewers performed 2,856 reviews of 1,484 separate manuscripts during a four-year study period, all prospectively rated on a standardized quality scale by editors. Multivariable analysis revealed that most variables, including academic rank, formal training in critical appraisal or statistics, or status as principal investigator of a grant, failed to predict performance of higher-quality reviews. The only significant predictors of quality were working in a university-operated hospital versus other teaching environment and relative youth (under ten years of experience after finishing training). Being on an editorial board and doing formal grant (study section) review were each predictors for only one of our two comparisons. However, the predictive power of all variables was weak. CONCLUSIONS: Our study confirms that there are no easily identifiable types of formal training or experience that predict reviewer performance. Skill in scientific peer review may be as ill defined and hard to impart as is "common sense." Without a better understanding of those skills, it seems unlikely journals and editors will be successful in systematically improving their selection of reviewers. This inability to predict performance makes it imperative that all but the smallest journals implement routine review ratings systems to routinely monitor the quality of their reviews (and thus the quality of the science they publish). PMID- 17411315 TI - Eight Americas: differences in Asian communities are important. PMID- 17411316 TI - Eight Americas: a new definition for "Americas"? PMID- 17411317 TI - HIV and cardiovascular disease: contribution of HIV-infected macrophages to development of atherosclerosis. PMID- 17411319 TI - HIV, stigma, and rates of infection: more complicated than Reidpath and Chan suggest. PMID- 17411320 TI - HIV, stigma, and rates of infection: a human rights and public health imperative. PMID- 17411321 TI - HIV, stigma, and rates of infection: but is AIDS-related stigma understood? PMID- 17411322 TI - HIV, stigma, and rates of infection: absence of evidence. PMID- 17411323 TI - Mental health in the millennium development goals: not ignored. PMID- 17411325 TI - Peer review in PLoS Medicine. PMID- 17411326 TI - Malaria in pregnancy: what can the social sciences contribute? PMID- 17411332 TI - The whole side of it--an interview with Neil Risch by Jane Gitschier. PMID- 17411327 TI - Biocurators: contributors to the world of science. PMID- 17411333 TI - Introducing PLoS Genetics. PMID- 17411335 TI - Psychophysical tests of the hypothesis of a bottom-up saliency map in primary visual cortex. AB - A unique vertical bar among horizontal bars is salient and pops out perceptually. Physiological data have suggested that mechanisms in the primary visual cortex (V1) contribute to the high saliency of such a unique basic feature, but indicated little regarding whether V1 plays an essential or peripheral role in input-driven or bottom-up saliency. Meanwhile, a biologically based V1 model has suggested that V1 mechanisms can also explain bottom-up saliencies beyond the pop out of basic features, such as the low saliency of a unique conjunction feature such as a red vertical bar among red horizontal and green vertical bars, under the hypothesis that the bottom-up saliency at any location is signaled by the activity of the most active cell responding to it regardless of the cell's preferred features such as color and orientation. The model can account for phenomena such as the difficulties in conjunction feature search, asymmetries in visual search, and how background irregularities affect ease of search. In this paper, we report nontrivial predictions from the V1 saliency hypothesis, and their psychophysical tests and confirmations. The prediction that most clearly distinguishes the V1 saliency hypothesis from other models is that task irrelevant features could interfere in visual search or segmentation tasks which rely significantly on bottom-up saliency. For instance, irrelevant colors can interfere in an orientation-based task, and the presence of horizontal and vertical bars can impair performance in a task based on oblique bars. Furthermore, properties of the intracortical interactions and neural selectivities in V1 predict specific emergent phenomena associated with visual grouping. Our findings support the idea that a bottom-up saliency map can be at a lower visual area than traditionally expected, with implications for top-down selection mechanisms. PMID- 17411336 TI - Integration of genome and chromatin structure with gene expression profiles to predict c-MYC recognition site binding and function. AB - The MYC genes encode nuclear sequence specific-binding DNA-binding proteins that are pleiotropic regulators of cellular function, and the c-MYC proto-oncogene is deregulated and/or mutated in most human cancers. Experimental studies of MYC binding to the genome are not fully consistent. While many c-MYC recognition sites can be identified in c-MYC responsive genes, other motif matches-even experimentally confirmed sites-are associated with genes showing no c-MYC response. We have developed a computational model that integrates multiple sources of evidence to predict which genes will bind and be regulated by MYC in vivo. First, a Bayesian network classifier is used to predict those c-MYC recognition sites that are most likely to exhibit high-occupancy binding in chromatin immunoprecipitation studies. This classifier incorporates genomic sequence, experimentally determined genomic chromatin acetylation islands, and predicted methylation status from a computational model estimating the likelihood of genomic DNA methylation. We find that the predictions from this classifier are also applicable to other transcription factors, such as cAMP-response element binding protein, whose binding sites are sensitive to DNA methylation. Second, the MYC binding probability is combined with the gene expression profile data from nine independent microarray datasets in multiple tissues. Finally, we may consider gene function annotations in Gene Ontology to predict the c-MYC targets. We assess the performance of our prediction results by comparing them with the c myc targets identified in the biomedical literature. In total, we predict 460 likely c-MYC target genes in the human genome, of which 67 have been reported to be both bound and regulated by MYC, 68 are bound by MYC, and another 80 are MYC regulated. The approach thus successfully identifies many known c-MYC targets and suggests many novel sites. Our findings suggest that to identify c-MYC genomic targets, integration of different data sources helps to improve the accuracy. PMID- 17411337 TI - Towards complete sets of farnesylated and geranylgeranylated proteins. AB - Three different prenyltransferases attach isoprenyl anchors to C-terminal motifs in substrate proteins. These lipid anchors serve for membrane attachment or protein-protein interactions in many pathways. Although well-tolerated selective prenyltransferase inhibitors are clinically available, their mode of action remains unclear since the known substrate sets of the various prenyltransferases are incomplete. The Prenylation Prediction Suite (PrePS) has been applied for large-scale predictions of prenylated proteins. To prioritize targets for experimental verification, we rank the predictions by their functional importance estimated by evolutionary conservation of the prenylation motifs within protein families. The ranked lists of predictions are accessible as PRENbase (http://mendel.imp.univie.ac.at/sat/PrePS/PRENbase) and can be queried for verification status, type of modifying enzymes (anchor type), and taxonomic distribution. Our results highlight a large group of plant metal-binding chaperones as well as several newly predicted proteins involved in ubiquitin mediated protein degradation, enriching the known functional repertoire of prenylated proteins. Furthermore, we identify two possibly prenylated proteins in Mimivirus. The section HumanPRENbase provides complete lists of predicted prenylated human proteins-for example, the list of farnesyltransferase targets that cannot become substrates of geranylgeranyltransferase 1 and, therefore, are especially affected by farnesyltransferase inhibitors (FTIs) used in cancer and anti-parasite therapy. We report direct experimental evidence verifying the prediction of the human proteins Prickle1, Prickle2, the BRO1 domain-containing FLJ32421 (termed BROFTI), and Rab28 (short isoform) as exclusive farnesyltransferase targets. We introduce PRENbase, a database of large-scale predictions of protein prenylation substrates ranked by evolutionary conservation of the motif. Experimental evidence is presented for the selective farnesylation of targets with an evolutionary conserved modification site. PMID- 17411338 TI - ART suppresses plasma HIV-1 RNA to a stable set point predicted by pretherapy viremia. AB - Current antiretroviral therapy is effective in suppressing but not eliminating HIV-1 infection. Understanding the source of viral persistence is essential for developing strategies to eradicate HIV-1 infection. We therefore investigated the level of plasma HIV-1 RNA in patients with viremia suppressed to less than 50-75 copies/ml on standard protease inhibitor- or non-nucleoside reverse transcriptase inhibitor-containing antiretroviral therapy using a new, real-time PCR-based assay for HIV-1 RNA with a limit of detection of one copy of HIV-1 RNA. Single copy assay results revealed that >80% of patients on initial antiretroviral therapy for 60 wk had persistent viremia of one copy/ml or more with an overall median of 3.1 copies/ml. The level of viremia correlated with pretherapy plasma HIV-1 RNA but not with the specific treatment regimen. Longitudinal studies revealed no significant decline in the level of viremia between 60 and 110 wk of suppressive antiretroviral therapy. These data suggest that the persistent viremia on current antiretroviral therapy is derived, at least in part, from long lived cells that are infected prior to initiation of therapy. PMID- 17411339 TI - M. tuberculosis Ser/Thr protein kinase D phosphorylates an anti-anti-sigma factor homolog. AB - Receptor Ser/Thr protein kinases are candidates for sensors that govern developmental changes and disease processes of Mycobacterium tuberculosis (Mtb), but the functions of these kinases are not established. Here, we show that Mtb protein kinase (Pkn) D overexpression alters transcription of numerous bacterial genes, including Rv0516c, a putative anti-anti-sigma factor, and genes regulated by sigma factor F. The PknD kinase domain directly phosphorylated Rv0516c, but no other sigma factor regulator, in vitro. In contrast, the purified PknB and PknE kinase domains phosphorylated distinct sigma regulators. Rather than modifying a consensus site, PknD phosphorylated Rv0516c in vitro and in vivo on Thr2 in a unique N-terminal extension. This phosphorylation inhibited Rv0516c binding in vitro to a homologous anti-anti-sigma factor, Rv2638. These results support a model in which signals transmitted through PknD alter the transcriptional program of Mtb by stimulating phosphorylation of a sigma factor regulator at an unprecedented control site. PMID- 17411340 TI - TGF-beta signaling controls embryo development in the parasitic flatworm Schistosoma mansoni. AB - Over 200 million people have, and another 600 million are at risk of contracting, schistosomiasis, one of the major neglected tropical diseases. Transmission of this infection, which is caused by helminth parasites of the genus Schistosoma, depends upon the release of parasite eggs from the human host. However, approximately 50% of eggs produced by schistosomes fail to reach the external environment, but instead become trapped in host tissues where pathological changes caused by the immune responses to secreted egg antigens precipitate disease. Despite the central importance of egg production in transmission and disease, relatively little is understood of the molecular processes underlying the development of this key life stage in schistosomes. Here, we describe a novel parasite-encoded TGF-beta superfamily member, Schistosoma mansoni Inhibin/Activin (SmInAct), which is key to this process. In situ hybridization localizes SmInAct expression to the reproductive tissues of the adult female, and real-time RT-PCR analyses indicate that SmInAct is abundantly expressed in ovipositing females and the eggs they produce. Based on real-time RT-PCR analyses, SmInAct transcription continues, albeit at a reduced level, both in adult worms isolated from single sex infections, where reproduction is absent, and in parasites from IL-7R(-/-) mice, in which viable egg production is severely compromised. Nevertheless, Western analyses demonstrate that SmInAct protein is undetectable in parasites from single-sex infections and from infections of IL-7R(-/-) mice, suggesting that SmInAct expression is tightly linked to the reproductive potential of the worms. A crucial role for SmInAct in successful embryogenesis is indicated by the finding that RNA interference-mediated knockdown of SmInAct expression in eggs aborts their development. Our results demonstrate that TGF-beta signaling plays a major role in the embryogenesis of a metazoan parasite, and have implications for the development of new strategies for the treatment and prevention of an important and neglected human disease. PMID- 17411341 TI - A dinucleotide deletion in CD24 confers protection against autoimmune diseases. AB - It is generally believed that susceptibility to both organ-specific and systemic autoimmune diseases is under polygenic control. Although multiple genes have been implicated in each type of autoimmune disease, few are known to have a significant impact on both. Here, we investigated the significance of polymorphisms in the human gene CD24 and the susceptibility to multiple sclerosis (MS) and systemic lupus erythematosus (SLE). We used cases/control studies to determine the association between CD24 polymorphism and the risk of MS and SLE. In addition, we also considered transmission disequilibrium tests using family data from two cohorts consisting of a total of 150 pedigrees of MS families and 187 pedigrees of SLE families. Our analyses revealed that a dinucleotide deletion at position 1527 approximately 1528 (P1527(del)) from the CD24 mRNA translation start site is associated with a significantly reduced risk (odds ratio = 0.54 with 95% confidence interval = 0.34-0.82) and delayed progression (p = 0.0188) of MS. Among the SLE cohort, we found a similar reduction of risk with the same polymorphism (odds ratio = 0.38, confidence interval = 0.22-0.62). More importantly, using 150 pedigrees of MS families from two independent cohorts and the TRANSMIT software, we found that the P1527(del) allele was preferentially transmitted to unaffected individuals (p = 0.002). Likewise, an analysis of 187 SLE families revealed the dinucleotide-deleted allele was preferentially transmitted to unaffected individuals (p = 0.002). The mRNA levels for the dinucleotide-deletion allele were 2.5-fold less than that of the wild-type allele. The dinucleotide deletion significantly reduced the stability of CD24 mRNA. Our results demonstrate that a destabilizing dinucleotide deletion in the 3' UTR of CD24 mRNA conveys significant protection against both MS and SLE. PMID- 17411343 TI - CTG trinucleotide repeat "big jumps": large expansions, small mice. AB - Trinucleotide repeat expansions are the genetic cause of numerous human diseases, including fragile X mental retardation, Huntington disease, and myotonic dystrophy type 1. Disease severity and age of onset are critically linked to expansion size. Previous mouse models of repeat instability have not recreated large intergenerational expansions ("big jumps"), observed when the repeat is transmitted from one generation to the next, and have never attained the very large tract lengths possible in humans. Here, we describe dramatic intergenerational CTG*CAG repeat expansions of several hundred repeats in a transgenic mouse model of myotonic dystrophy type 1, resulting in increasingly severe phenotypic and molecular abnormalities. Homozygous mice carrying over 700 trinucleotide repeats on both alleles display severely reduced body size and splicing abnormalities, notably in the central nervous system. Our findings demonstrate that large intergenerational trinucleotide repeat expansions can be recreated in mice, and endorse the use of transgenic mouse models to refine our understanding of triplet repeat expansion and the resulting pathogenesis. PMID- 17411342 TI - Generalized analysis of molecular variance. AB - Many studies in the fields of genetic epidemiology and applied population genetics are predicated on, or require, an assessment of the genetic background diversity of the individuals chosen for study. A number of strategies have been developed for assessing genetic background diversity. These strategies typically focus on genotype data collected on the individuals in the study, based on a panel of DNA markers. However, many of these strategies are either rooted in cluster analysis techniques, and hence suffer from problems inherent to the assignment of the biological and statistical meaning to resulting clusters, or have formulations that do not permit easy and intuitive extensions. We describe a very general approach to the problem of assessing genetic background diversity that extends the analysis of molecular variance (AMOVA) strategy introduced by Excoffier and colleagues some time ago. As in the original AMOVA strategy, the proposed approach, termed generalized AMOVA (GAMOVA), requires a genetic similarity matrix constructed from the allelic profiles of individuals under study and/or allele frequency summaries of the populations from which the individuals have been sampled. The proposed strategy can be used to either estimate the fraction of genetic variation explained by grouping factors such as country of origin, race, or ethnicity, or to quantify the strength of the relationship of the observed genetic background variation to quantitative measures collected on the subjects, such as blood pressure levels or anthropometric measures. Since the formulation of our test statistic is rooted in multivariate linear models, sets of variables can be related to genetic background in multiple regression-like contexts. GAMOVA can also be used to complement graphical representations of genetic diversity such as tree diagrams (dendrograms) or heatmaps. We examine features, advantages, and power of the proposed procedure and showcase its flexibility by using it to analyze a wide variety of published data sets, including data from the Human Genome Diversity Project, classical anthropometry data collected by Howells, and the International HapMap Project. PMID- 17411344 TI - Integration of light and temperature in the regulation of circadian gene expression in Drosophila. AB - Circadian clocks are aligned to the environment via synchronizing signals, or Zeitgebers, such as daily light and temperature cycles, food availability, and social behavior. In this study, we found that genome-wide expression profiles from temperature-entrained flies show a dramatic difference in the presence or absence of a thermocycle. Whereas transcript levels appear to be modified broadly by changes in temperature, there is a specific set of temperature-entrained circadian mRNA profiles that continue to oscillate in constant conditions. There are marked differences in the biological functions represented by temperature driven or circadian regulation. The set of temperature-entrained circadian transcripts overlaps significantly with a previously defined set of transcripts oscillating in response to a photocycle. In follow-up studies, all thermocycle entrained circadian transcript rhythms also responded to light/dark entrainment, whereas some photocycle-entrained rhythms did not respond to temperature entrainment. Transcripts encoding the clock components Period, Timeless, Clock, Vrille, PAR-domain protein 1, and Cryptochrome were all confirmed to be rhythmic after entrainment to a daily thermocycle, although the presence of a thermocycle resulted in an unexpected phase difference between period and timeless expression rhythms at the transcript but not the protein level. Generally, transcripts that exhibit circadian rhythms both in response to thermocycles and photocycles maintained the same mutual phase relationships after entrainment by temperature or light. Comparison of the collective temperature- and light-entrained circadian phases of these transcripts indicates that natural environmental light and temperature cycles cooperatively entrain the circadian clock. This interpretation is further supported by comparative analysis of the circadian phases observed for temperature-entrained and light-entrained circadian locomotor behavior. Taken together, these findings suggest that information from both light and temperature is integrated by the transcriptional clock mechanism in the adult fly head. PMID- 17411345 TI - Lifespan regulation by evolutionarily conserved genes essential for viability. AB - Evolutionarily conserved mechanisms that control aging are predicted to have prereproductive functions in order to be subject to natural selection. Genes that are essential for growth and development are highly conserved in evolution, but their role in longevity has not previously been assessed. We screened 2,700 genes essential for Caenorhabditis elegans development and identified 64 genes that extend lifespan when inactivated postdevelopmentally. These candidate lifespan regulators are highly conserved from yeast to humans. Classification of the candidate lifespan regulators into functional groups identified the expected insulin and metabolic pathways but also revealed enrichment for translation, RNA, and chromatin factors. Many of these essential gene inactivations extend lifespan as much as the strongest known regulators of aging. Early gene inactivations of these essential genes caused growth arrest at larval stages, and some of these arrested animals live much longer than wild-type adults. daf-16 is required for the enhanced survival of arrested larvae, suggesting that the increased longevity is a physiological response to the essential gene inactivation. These results suggest that insulin-signaling pathways play a role in regulation of aging at any stage in life. PMID- 17411346 TI - Reproductive toxicity testing under the REACH system--time for a paradigm shift. PMID- 17411347 TI - Comparative quantitative structure-activity-activity relationships for toxicity to Tetrahymena pyriformis and Pimephales promelas. AB - An approach for predicting acute aquatic toxicity, in the form of a quantitative structure-activity-activity relationship (QSAAR), is described. This study assessed relative toxic effects to a fish, Pimephales promelas, and a ciliate, Tetrahymena pyriformis, and attempted to form relationships between them. A good agreement between toxic potencies (R2 = 0.754) was found for a chemically diverse dataset of 364 compounds, when using toxicity to the ciliate as a surrogate to that for fish. This relationship was extended by adding three theoretical structural descriptors of the molecules. The inclusion of these descriptors improved the relationship further (R2 = 0.824). The structural features that were found to improve the extrapolation between the toxicity to the two different species were related to the electron distribution of the carbon skeleton of the toxicant, its hydrogen-bonding ability, and its relative nitrogen content. Such a QSAAR approach provides a potential tool for predicting the toxicities of chemicals for environmental risk assessment and thus for reducing animal tests. PMID- 17411348 TI - A comparative study of machine learning algorithms applied to predictive toxicology data mining. AB - This paper reports results of a comparative study of widely used machine learning algorithms applied to predictive toxicology data mining. The machine learning algorithms involved were chosen in terms of their representability and diversity, and were extensively evaluated with seven toxicity data sets which were taken from real-world applications. Some results based on visual analysis of the correlations of different descriptors to the class values of chemical compounds, and on the relationships of the range of chosen descriptors to the performance of machine learning algorithms, are emphasised from our experiments. Some interesting findings relating to the data and the quality of the models are presented--for example, that no specific algorithm appears best for all seven toxicity data sets, and that up to five descriptors are sufficient for creating classification models for each toxicity data set with good accuracy. We suggest that, for a specific data set, model accuracy is affected by the feature selection method and model development technique. Models built with too many or too few descriptors are undesirable, and finding the optimal feature subset appears at least as important as selecting appropriate algorithms with which to build a final model. PMID- 17411349 TI - The integrated acute systemic toxicity project (ACuteTox) for the optimisation and validation of alternative in vitro tests. AB - The ACuteTox project is designed to replace animal testing for acute systemic toxicity, as is widely used today for regulatory purposes, by using in vitro and in silico alternatives. In spite of the fact that earlier studies on acute systemic toxicity demonstrated a good correlation between in vitro basal cytotoxicity data (the 50% inhibitory concentration [IC50]) in human cell lines and rodent LD50 values, and an even better correlation between IC50 values and human lethal blood concentrations, very few non-animal tests have been accepted for general use. Therefore, the aim of the ACuteTox project is to adapt new testing strategies, for example, the implementation of new endpoints and new cell systems for toxicity screening, organ-specific models, metabolism-dependent toxicity, tissue absorption, distribution and excretion, and computer-based prediction models. A new database, AcuBase, containing descriptions and results of in vitro tests of the 97 reference chemicals, as well as the results of animal experimentation, and human acute toxicity data, will be generated within the framework of ACuteTox. Scientists from 13 European countries are working together and making efforts to find the most appropriate testing strategies for the prediction of human acute systemic toxicity, and also to select a robust in vitro test battery for cytotoxicity testing of chemicals. PMID- 17411350 TI - The improvement of in vitro cytotoxicity testing for the assessment of acute toxicity in fish. AB - The use of fish cell line cytotoxicity tests as alternatives to acute lethality tests with fish is hampered by the clearly lower sensitivity of the fish cell line tests. Recently, it has been shown that this is not a unique feature of fish cells. In fact, the sensitivity of mammalian and human cell lines toward the cytotoxic actions of chemicals, in general, is comparable to that of fish cell lines. Reviewing some of our recent investigations, the objective of this paper is to show that the sensitivity of in vitro cytotoxicity testing and the correspondence between in vitro cytotoxic and acute fish toxic concentrations (LC50) can be increased, if: a) inhibition of cell growth instead of cell death is used as the endpoint; and b) the bioavailable free cytotoxic concentration (ECu50) of chemicals in vitro, instead of the nominal cytotoxic concentration (EC50), is used as the measure of cytotoxic potency. Based on these results, a pragmatic in vitro testing strategy for estimating the minimal aquatic toxic potency of chemicals is proposed. PMID- 17411351 TI - An in vitro study of the toxic effects of Stachybotrys chartarum metabolites on lung cells. AB - During a study of indoor fungal colonisation, several isolates of Stachybotrys chartarum were recovered, and the effects of metabolites from four isolates on lung epithelial Type II cells and alveolar macrophages were studied in vitro. All the isolates showed toxic effects on both cell types, and they differed only in the extent of the changes induced. In Type II cells, the number of alkaline phosphatase positive cells was reduced, the pattern of Maclura pomifera agglutinin (MPA) binding was changed, and acid phosphatase activity in alveolar macrophages was diminished. In both cell types, the production of monocyte chemotactic protein-1 (MCP-1) and tumour necrosis factor-alpha (TNF-alpha) was changed, and parameters related to antioxidant status (superoxide dismutase, glutathione peroxidase, glutathione) were decreased. PMID- 17411352 TI - Induction of apoptosis and activation of JNK and p38 MAPK pathways in deoxynivalenol-treated cell lines. AB - Deoxynivalenol (DON) is a mycotoxin produced by what are thought to be the most prevalent toxin-producing fungi of the Fusarium genus. Here, we present the results of apoptosis induction, phosphorylation of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases (MAPKs), and expression of the c-Jun protein after DON treatment, in a pre-B lymphocyte REH cell line. In addition, human pre-T lymphocyte Jurkat, hamster kidney-derived BHK21 and mouse hepatoma MH 22a cells were used in comparative experiments in vitro. We found that the DON effect was cell origin-dependent and dose-dependent, with a significant slow-down of cell proliferation and increase of apoptotic cells in blood cell lines. BHK21 and MH-22a cells were less sensitive to the DON effect. In blood-derived REH and Jurkat cells, DON-induced apoptotic changes were preceded by an increase in JNK and p38 MAPKs phosphorylation, as well as in c-Jun expression. However, the activation of JNK phosphorylation and c-Jun expression were transient, but did not coincide with each other. An inhibitor of JNK1/2, SP600125, had a negligible negative effect on REH cell viability after DON treatment, demonstrating that JNK does not contribute to DON-induced apoptosis. In contrast, studies on the role of p38 MAPK revealed that p38 signalling is required for DON-induced apoptosis in REH cells. PMID- 17411353 TI - The developmental expression of fluorescent proteins in organotypic hippocampal slice cultures from transgenic mice and its use in the determination of excitotoxic neurodegeneration. AB - Transgenic mice, expressing fluorescent proteins in neurons and glia, provide new opportunities for real-time microscopic monitoring of degenerative and regenerative structural changes. We have previously validated and compared a number of quantifiable markers for neuronal damage and cell death in organotypic brain slice cultures, such as cellular uptake of propidium iodide (PI), loss of microtubule-associated protein 2 (MAP2), Fluoro-Jade (FJ) cell staining, and the release of cytosolic lactate dehydrogenase (LDH). An important supplement to these markers would be data on corresponding morphological changes, as well as the opportunity to monitor reversible changes or long-term effects in the event of minor damage. As a first step, we present: a) the developmental expression in organotypic hippocampal brain slice cultures of transgenic fluorescent proteins, useful for the visualisation of neuronal subpopulations and astroglial cells; and b) examples of excitotoxic, glutamate receptor-induced degeneration of hippocampal CA1 pyramidal cells, with corresponding astroglial reactivity in such cultures. The slice cultures were set up according to standard techniques, by using one-week old pups from four transgenic mouse strains which express fluorescent proteins in their neurons and/or astroglial cells. From the time of explantation, and subsequently for up to nine weeks in culture, the transgenic neuronal fluorescence displayed the expected characteristics of a developmental, in vivo-like increase, including both the number and localisation of cells, as well as the intensity of fluorescence. At that stage and later, the transgenic fluorescence clearly permitted the visualisation of cell bodies, larger and smaller dendritic branches, spines and axons. In separate experiments, with a 24 hour exposure of matured sliced cultures to 100 microM of the glutamate agonist, N-methyl-D-aspartate (NMDA), we observed, by time-lapse recording, a gradual, but rapid loss of fluorescent CA1 pyramidal cells, accompanied by astrogliosis of transgene fluorescent astroglial cells. Based on these results, we consider that organotypic brain slice cultures from transgenic mice, with fluorescent neurons and glia, combined with detailed visualisation by time-lapse fluorescence microscopy, have great potential for investigating both major irreversible and minor reversible structural changes in neurons and glia, induced by neurotoxins and other neurodegenerative compounds and conditions. PMID- 17411354 TI - The effects of selected phenol and phthalate derivatives on steroid hormone production by cultured porcine granulosa cells. AB - We have investigated the effects of several phenols (octylphenol [OP], nonylphenol [NP], tert-octylphenol [tOP]) and phthalates (dioctylphthalate [DOP], diisodecylphthalate [DiDP], diisononylphthalate [DiNP]) on steroid hormone production by porcine ovarian granulosa cells after a 72-hour incubation. These chemicals are widely used as plasticisers and are suspected to possess endocrine disrupting properties. No changes were exhibited in basal progesterone production after treatment with NP or tOP, or with the tested phthalates. However, OP tended to decrease progesterone levels, while DOP and DiDP, at the lowest concentration used (10(-8)M), increased progesterone levels in the culture media. Neither of the tested phenols affected follicle stimulating hormone (FSH)-stimulated progesterone production, except for OP and NP at 10(-4)M, which decreased progesterone levels. The phthalates, tested at higher concentrations, were able to amplify FSH-stimulated progesterone release into the culture medium. An inhibitory action on oestradiol production by porcine granulosa cells was observed after the treatment with both groups of test chemicals. The results obtained in the experiments on primary granulosa cell cultures indicate that ovarian steroidogenesis might be one of the possible sites affected by the endocrine disrupting actions of phenols and phthalates. PMID- 17411355 TI - Cyclosporin A potentiates the cytotoxic effects of methyl methanesulphonate in HL 60 and K562 cells. AB - Methyl methanesulphonate (MMS) is a DNA damaging agent, which induces oxidative stress, ATP depletion, and consequently, cell death, in HL-60 and K562 cells. The cell death induced by MMS predominantly exhibited the morphological and biochemical hallmarks of necrosis. A minor population of dying cells exhibited apoptotic hallmarks, especially in K562 cell cultures. Cyclosporin A (CsA) was used to modulate the MMS-induced cell death. Our results indicated that CsA did not prevent cells from dying, but changed the mode of death from necrotic to apoptotic. Surprisingly, CsA enhanced oxidative stress and increased the overall number of dead cells. Based on these results, we conclude that the modulatory effect of CsA on MMS-induced cell death might arise from an interference by CsA with mitochondrial metabolism, rather than from inhibition of the MMS efflux mediated by P-glycoprotein. PMID- 17411356 TI - The effects of heavy metals on common carp white blood cells in vitro. AB - The in vitro effects of cadmium, copper, lead and zinc, and various cadmium compounds (chloride, sulphate and nitrate) on common carp (Cyprinus carpio) lymphocyte viability and phagocyte activity, were evaluated. The percentage of dead lymphocytes was determined after Trypan blue staining, and phagocyte activity was measured by using the nitroblue tetrazolium (NBT) reduction test. Lead was the most toxic to lymphocytes--the maximum mortality exceeded 30%, and was significantly higher at 1 microM of lead, compared to the control. The maximum mortality caused by cadmium was below 10%, but was significantly elevated with 5 microM or more of cadmium. Zinc induced lymphocyte mortality from 10 microM, whilst no effect was observed with copper. The incubation of full blood with the three cadmium compounds (at 5mg/l of cadmium) showed that cadmium nitrate and cadmium sulphate were more toxic (over 35% and 25% mortality, respectively) than cadmium chloride (about 15% mortality). This was confirmed by the results of tests on isolated cells--1mg/l of cadmium as nitrate and sulphate increased lymphocyte mortality compared to the control and cadmium chloride. Phagocytic activity was less sensitive to heavy metals than was lymphocyte viability. It was significantly reduced following exposure to 50 microM and 100 microM cadmium, and 100 microM zinc, but no effects were observed with either lead or copper. PMID- 17411357 TI - Environmental and synthetic sulphydryl group inhibitors: effects on bioluminescence and respiration in Vibrio fischeri. AB - Elemental sulphur (as S0 and S8) is abundant in anaerobic sediments and soil, and is highly toxic in the Vibrio fischeri bioluminescence test. This mode of S0 action remains uncertain. The objective of this research was the analysis of the toxic effects of S0 on bioluminescence and respiration in V. fischeri, in joint action with N-ethylmaleimide (NEM) or 2,4-dithio-DL-threitol (DTT), which are -SH group inhibiting and maintaining synthetic agents, respectively. Non-toxic DTT immediately protected cell bioluminescence against S0 inhibition at low (5.5ppb) and high (55ppb) concentrations of S0, whilst restoration of the inhibitory effect of S0 took up to 30 minutes. NEM (62.5ppb) diminished cell bioluminescence by up to 50% after 5 minutes, but after 60 minutes, the inhibition reached 100%. DTT restored the bioluminescence function inhibited in vivo and in vitro by S0 and NEM. Enhancement of cell respiration by up to 20% and 33% was observed at 2.2ppm of S0 and 36.8ppm of 2,4-dinitrophenol (2,4-DNP; an uncoupler of oxidative phosphorylation), respectively; whilst NEM (3.1ppm) caused a reduction of up to 40%. This comparative analysis confirmed that S0 has multiple modes of action--it acts as both an -SH group inhibitor and an uncoupler of oxidative phosphorylation in V. fischeri cells. PMID- 17411358 TI - Rapid screening for soil ecotoxicity with a battery of luminescent bacteria tests. AB - A bacterial test battery, involving i) Microtox, an aquatic test, ii) the Flash assay, a soil-suspension test (with Vibrio fischeri as the test organism), and iii) the Metal Detector assay, a semi-specific aquatic test for heavy metals (with recombinant luminescent Escherichia coli), was used in a combined toxicological and chemical hazard assessment of Estonian soils sampled from a former Soviet military airfield (13 samples) and from traffic-influenced roadsides (5 samples). The soils showed slightly elevated levels of total petroleum hydrocarbons (TPH), but not of heavy metals. In most of the samples, the levels of TPH did not exceed the Estonian permitted limit values set for residential areas. Toxicity testing was performed on both fresh and dried soils, after aqueous extraction for 1 hour and 24 hours. The toxicity results obtained with the Microtox test did not significantly differ in all of the sample treatment schemes; however, it appeared that the drying and sieving of the soils increased the bioavailability of toxicants, probably due to an enlarged reactive soil surface area. According to chemical analysis of the soils and the data from the Microtox test and the Metal Detector assay (performed on aqueous elutriates of the soils), these soils would not be considered to be hazardous. In contrast, the Flash assay performed on soil-water suspensions of dried soils, showed that most of the soils were toxic and thus probably contained undetermined particle bound bioavailable toxicants. The photobacterial toxicity test (the Flash assay) can be recommended for the rapid screening of soils, as it is sensitive, cheap and inexpensive, and provides valuable information on particle-bound bioavailable toxicants, useful for complementing a chemical analysis and for assessing the risks originating from polluted soils. PMID- 17411359 TI - The use of bioassays for the risk assessment of toxic leachates: an experimental study. AB - Solid wastes from the oil-shale industry produce leachates containing toxic compounds such as heavy metals and persistent polycyclic aromatic hydrocarbons (PAH). The hazard to the environment represented by waste leachates depends not only on their chemical composition, but also on the mobility and bioavailability of toxic contaminants in soils. We evaluated the applicability of bioassays for toxicity assessment of the bioavailable fraction of heavy metals and PAH in soils, in experiments with samples of four different soil types (Rendzina, Brown pseudopodzolic, Typical brown, Sodpodzolic), the pH of which ranged from 6.2 to 7.2. The toxicity of the bioavailable fraction of the soil contaminants was assessed with the dehydrogenase enzyme activity assay, and with a Toxkit microbiotest with the crustacean, Thamnocephalus platyurus, after treatment of the soil samples with an artificial solution containing chromium (III), lead (II), copper (II), cadmium (II) and pyrene. The test results confirm those of earlier experiments, which characterised the sorption potential of investigated soils for the same compounds. Both tests turned out to be sufficiently sensitive, and hence can be recommended as effective and useful tools for the assessment of the bioavailable fraction of soil contaminants. PMID- 17411360 TI - The role of laboratory and field leaching tests in hazard identification for solid materials. AB - The use of various in vitro toxicity assays for testing environmental solid samples is dependent on the availability of reliable methods for the sampling and pretreatment of the material. This study focuses on the evaluation of leaching behaviour as a first step in the context of the toxicity testing of solid environmental matter. Spent shale, from oil shale retorting, was chosen as a suitable example of deposited solid waste material. For the generation of leachate in the laboratory setting, a standard two-stage batch-leaching test was applied to the samples of technogenic waste. In the field, a new type of lysimeter, which does not disturb the surface, was used for in situ leachate collection. The chemical composition of water extracts was found to be different under field conditions, as compared with the laboratory experiments. Thus, the hazard identification of a solid technogenic waste by in vitro toxicological tests applied to laboratory leachates would not be the best solution. The content of hazardous ingredients could be underestimated if only laboratory tests are used. For risk assessment concerned with solid waste materials, the generation of leachate by using field lysimeters is recommended. PMID- 17411361 TI - Conformational changes in proteins in vitro as a means of predicting the acute toxicities of chemicals. AB - Experimental data are presented on ovalbumin denaturation (OD, EC10) and human acetylcholine esterase (AChE) inhibition (IC50) in vitro, following exposure to the chemicals used in the international Multicentre Evaluation of In vitro Cytotoxicity (MEIC) programme. Data were obtained for 40 (OD test) and 43 (AChE test) of the 50 MEIC chemicals. These data were compared with similar data from other methods used in the MEIC programme, and good correlations (R2) were obtained with data from MEIC studies on cell lines: 0.80 for human, 0.81 for other animal, and 0.78 for fish cell line IC50 values and AChE values, and 0.76 for human, 0.69 other animal and 0.75 for fish cell line IC50 values and OD values. The correlation increased substantially, if chemicals which freely cross the blood-brain barrier were solely considered, with R2 = 0.90 for human, 0.90 for other animal, and 0.82 for fish cell line IC50 values and AchE values, and 0.87 for human, 0.86 for other animal, and 0.92 for fish cell line IC50 values and OD values, in this case. Such chemicals are the main cause of non-specific depression of the central nervous system (CNS). The AChE IC50 permits a good prediction of human acute toxicity, similar to the IC50 values obtained with human cell lines and the same MEIC chemicals. These results confirm the basal toxicity hypothesis formulated by Bjorn Ekwall. It is concluded that in vitro methods based on the disruption of the functions of the proteins vital for body operation can be used as an alternative to the cell culture methods, when non specific toxic effects of chemicals on humans and animals are evaluated. PMID- 17411362 TI - Skin sensitisation and epidermal disposition: the relevance of epidermal disposition for sensitisation hazard identification and risk assessment. The report and recommendations of ECVAM workshop 59. PMID- 17411363 TI - Comments on the sub-group reports of the EU Technical Expert Working Group on the revision of Directive 86/609/EEC on the protection of animals used for experimental and other scientific purposes. AB - A critical analysis is presented of the reports produced by four Technical Expert Working Group Sub-groups (SGs) on Ethical Review, Cost-Benefit, Authorisation and Scope, which were published on the EC website (http://ec.europa.eu/environment/chemicals/lab_animals/ia_info_en.htm), as part of the European Commission (EC)s review of EU Directive 86/609 EEC. This is in addition to our official response to the internet consultation questionnaire, submitted to the Commission on behalf of FRAME. Whilst the respective SG reports were extensive and detailed, we have identified several shortcomings, and in particular, a frequent lack of consensus among the SG members, resulting in a lack of clear guidance for the EC on what the revised Directive should contain, with reference to a number of crucial issues. Such indecisiveness could lead to wide discrepancies in the approaches of the EC, the European Parliament and the EU Member States concerning many issues of importance to animal welfare and the implementation of alternatives to animal experiments. These concerns range from logistical issues, such as requirements for named officers in authorised establishments, and the recording and publishing of statistics on animal use, to ethical and scientific problems, including the use of non-human primates, local ethical review, and education and training on the essential link between the Three Rs concept and best scientific practice. In each case, the basis for our concerns is explained, and suggestions for improvements to be incorporated into the revised Directive are made, in the hope that the harmonisation of approaches to laboratory animal experimentation and the use of alternative methods in the Member States can be maximised. PMID- 17411364 TI - Why do the numbers of laboratory animal procedures conducted continue to rise? An analysis of the Home Office Statistics of Scientific Procedures on Living Animals: Great Britain 2005. AB - The publication of the Statistics of Scientific Procedures on Living Animals: Great Britain 2005 once again provides evidence that the levels of animal experimentation in Great Britain are rising, the underlying reason for this being the continued and increasing reliance on genetically modified animals as model systems. There has been a gradual increase in fundamental research, as applied toxicological studies have declined. Of particular concern is the impact that the forthcoming REACH legislation will have and the apparent lack of urgency in facing up to this challenge. The major issues arising from the Statistics are discussed, including the increases in rabbit and primate procedures. The potential of newly validated and emerging techniques to counteract these worrying trends are also considered. PMID- 17411365 TI - Extended core sequences from the cHS4 insulator are necessary for protecting retroviral vectors from silencing position effects. AB - The prototypic chromatin insulator cHS4 has proven effective at reducing repressive chromosomal position effects on retroviral vector expression. We report here studies designed to identify the minimal chicken hypersensitive site 4 (cHS4) sequences necessary for this activity. Using a gammaretroviral reporter vector and expression analysis in cell lines and primary mouse hematopoietic progenitor colonies, we found that a 250-bp core fragment reported to contain most of the cHS4 insulating activity failed to prevent silencing when used alone, although some barrier activity was observed when this fragment was combined with a 790-bp, but not 596-bp, spacer. Similar studies showed that four copies of a 90 bp fragment containing the cHS4 enhancer-blocking activity actually repressed vector green fluorescent protein (GFP) expression. In contrast, a 400-bp fragment containing the 250-bp core plus 3' flanking sequences protected vector expression to the same degree as the full-length 1.2-kb fragment. The 400-bp fragment activity was confirmed in a lentiviral vector expressing human beta-globin in murine erythroid leukemia (MEL) cells. Taken together, these studies indicate that the insulating activity of the 250-bp cHS4 core can be influenced by distance, and identify an extended core element that confers full barrier activity in the setting of two different classes of retroviral vectors. PMID- 17411366 TI - Regulating the functions of the HIV-1 matrix protein. AB - The HIV-1 structural protein matrix (MA) is involved in a number of essential steps during infection and appears to possess multiple, seemingly conflicting targeting signals. Although MA has long been known to be crucial for virion assembly, details regarding this function, and the domains responsible for mediating it, are still emerging. MA has also been implicated in nuclear import of HIV cDNA and is purported to contain a nuclear targeting signal. Little is known about how these opposing plasma membrane and nuclear targeting signals are regulated and which signals predominate at various stages of infection. Additionally, MA has recently been implicated in a number of novel roles during infection including viral entry/uncoating, cytoskeletal-mediated transport, and targeting viral assembly to lipid rafts. Here we discuss our current understanding of MA's functions during infection and explore the recent advancements made in elucidating the mechanism of these processes. It appears that MA possesses a cache of targeting signals that are likely to be regulated throughout the infectious cycle by a combination of structural and biochemical modifications including phosphorylation, myristoylation, and multimerization. The ability of HIV to modify the properties of MA at specific stages of infection is central to the multifunctional behavior of MA and the efficiency of HIV infection. The recently reported success of drugs specifically designed to block MA function (Haffar O, Dubrovsky L, and Lowe R et al. J Virol 2005;79:13028 13036) confirms the importance of this protein for HIV infection and highlights a potentially new avenue in multivalent drug therapy. PMID- 17411367 TI - Limitations to contact tracing and phylogenetic analysis in establishing HIV type 1 transmission networks in Cuba. AB - Sequence analysis can be used to evaluate transmission networks. We have used retrospective samples to examine two HIV-1 transmission networks established by contact tracing. Regions of the HIV-1 region representing segments of gag and env were amplified by RT-PCR from frozen plasma samples and the sequence of each PCR product was determined. Within one of the networks (composed of 38 subjects) we found only a subset of the tested sequence clusters was consistent with the reported epidemiological linkage. Of 15 presumed transmission events where sequence data were available, 9 could be rejected either by subtype mismatch or by phylogenetic tests. In the other network (composed of 89 subjects) we were able to assess sequences for 26 presumed transmission events, 18 of which were rejected based on subtype discordance. Long lags in time between the time of transmission and the time of sequence sampling (ranging from 2 to 18 years) may limit the sensitivity for the detection of sequence linkage. Also, superinfection and incomplete epidemiological information are other factors that will limit the concordance of phylogenetic reconstruction and reported epidemiological linkage. PMID- 17411368 TI - Genotypic resistance mutations to antiretroviral drugs in treatment-naive HIV/AIDS patients living in Liaoning Province, China: baseline prevalence and subtype-specific difference. AB - To examine the prevalence of drug resistance mutations among treatment-naive HIV/AIDS patients living in Liaoning province in China, the HIV-1 pol gene from plasma of 91 seropositive treatment-naive patients was amplified and sequenced. Three patients (3.3%) had an M46I amino acid substitution in the protease (PR) gene that decreased susceptibility to IDV, RTV, and NFV and one patient (1.1%) had an M184I amino acid substitution in the reverse transcriptase (RT) gene that confers high-level resistance to 3TC and FTC. Minor mutations were detected in high frequency in the PR gene. The frequencies of minor mutations to protease inhibitors (PI) were I93L (71.4%), L63P (62.6%), V77I (62.6%), M36I/V (33.0%), A71T/V (22.0%), K20R (6.6%), G16E (6.6%), and L10I (5.5%). The relatedness between subtypes and the frequencies of amino acid substitutions in PR were observed; 63P, 77I, and 71V/T were found in HIV-1 subtype B'/B, 16E, 36I, 20R/I, and 82I in non-B, except for CRF07_BC, 10I in subtype A, and 93L in non-A. Although the primary resistance of HIV-1 to antiretroviral drugs is low among the treatment-naive HIV-1 patients living in Liaoning province, the surveillance and monitoring of drug-resistant HIV-1 should be implemented regularly because of the increased access to antiretroviral therapy in China. PMID- 17411369 TI - Clinical manifestations associated with HTLV type I infection: a cross-sectional study. AB - Human T-lymphotropic virus type I (HTLV-I) causes HTLV-I-associated myelopathy/tropical spastic paraparesis and adult T cell leukemia in a small percentage of infected individuals. HTLV-I infection is increasingly associated with clinical manifestations. To determine the prevalence of clinical manifestations in HTLV-I infected individuals, we conducted a cross-sectional study of 115 HTLV-I-infected blood donors without myelopathy and 115 age- and sex matched seronegative controls. Subjects answered a standardized questionnaire and underwent physical examination. Compared with controls, HTLV-I-infected subjects were more likely to report arm or leg weakness (OR = 3.8, 95% CI: 1.4-10.2; OR = 4.0, 95% CI: 1.6-9.8, respectively), hand or foot numbness (OR = 2.1, 95% CI: 1.1 3.9; OR = 4.8, 95% CI: 2.0-11.7, respectively), arthralgia (OR = 3.3, 95% CI: 1.7 6.4), nocturia (OR = 2.7, 95% CI: 1.04-6.8), erectile dysfunction (OR = 4.0, 95% CI: 1.6-9.8), and to have gingivitis (OR = 3.8, 95% CI: 1.8-7.9), periodontitis (OR = 10.0, 95% CI: 2.3-42.8), and dry oral mucosa (OR = 7.5, 95% CI: 1.7-32.8). HTLV-I infection is associated with a variety of clinical manifestations, which may occur in patients who have not developed myelopathy. PMID- 17411370 TI - Contribution of monocytes to viral replication in macaques during acute infection with simian immunodeficiency virus. AB - Monocytes are known as an alternative target for HIV/SIV infection, but the contribution of monocytes to viral spread in a host is unclear. In this study, CD14 monocytes were monitored in 6 macaques until six weeks postinfection (wpi) with SIVmac239 to evaluate their contribution to viral load. The monocyte count in blood significantly increased with peak viremia at 2 wpi and the expression level of CD14 on monocytes significantly decreased at 1-2 wpi, though the number of CD4(+) T cells was stable in these macaques. The number of CD14 monocytes and the expression level of CD14 on monocytes at 2 wpi were also significantly related to the extent of viremia in plasma. An increased number of monocytes at 2 wpi was associated with a lower postacute viral load, suggesting that monocytes have a role in suppressing the virus. The lower expression level of CD14 in monocytes at 2 wpi was associated with a higher viral load and greater degree of infection of monocytes. This correlation suggests that monocytes with a low level of CD14 may be more susceptible to SIV and may enhance viral replication. The analysis of monocytes in persistently infected macaques revealed that the expression level of CD14 was also significantly low during persistent infection compared with naive macaques, though the monocyte count was within the normal range. Monocytes may suppress viruses, perhaps by their immune function, during acute infection. However, infection of monocytes may increase the viral load and spread viruses in a host. PMID- 17411371 TI - Slow human immunodeficiency virus type 1 evolution in viral reservoirs in infants treated with effective antiretroviral therapy. AB - A longitudinal study of viral reservoirs in children initiating highly active antiretroviral therapy (HAART) in early infancy was undertaken to test the hypothesis that early effective treatment affects the persistence of replication competent viral latency and the evolution of HIV-1 in resting CD4(+) T cells. An end point dilution culture assay was used to measure the frequencies of latently infected resting CD4(+) T cells harboring replication-competent virus in early and late treated children. Gag, pol, and env also were sequenced and compared to pretreatment sequences. HIV-1-specific humoral and cellular immune responses were also assessed. Blood samples were obtained from 12 HIV-1-infected children who started HAART at a median of 1.9 months of age and who maintained suppression of HIV-1 replication for up to 5.5 years. Replication-competent HIV-1 was recovered from 10/12 (84%) subjects. Evolution in gag, pol, and env was restricted for years in early-treated children. HAART initiated from early infancy does not prevent the establishment of a reservoir of latent provirus, but does significantly limit the evolution of HIV-1 in viral reservoirs. The effect of early therapy on HIV-1 evolution may have implications for long-term pharmacologic control of HIV-1. PMID- 17411372 TI - Vpr in plasma of HIV type 1-positive patients is correlated with the HIV type 1 RNA titers. AB - Vpr, an accessory gene product of HIV-1, has been reported in the plasma of HIV-1 positive patients, and exogenous Vpr induces the reactivation of viral production from latently infected cells and the apoptosis of T cells in vitro. These observations imply that Vpr is important in AIDS development, but the clinical relevance of the findings cannot be evaluated fully because the actual plasma Vpr concentration in HIV-1-positive patients is unknown. Here we generated two monoclonal antibodies against different portions of Vpr and successfully identified Vpr as a 14-kDa protein in HIV-1-positive patients. Semiquantitative analysis using a recombinant Vpr revealed that the concentration of Vpr in patient plasma was approximately 0.7 nM (10 ng/ml). Cross-sectional analysis of 52 HIV-1-positive patients revealed that the presence of Vpr detected in 20 patients was positively correlated with HIV-1 RNA copy number (p > 0.03), but not with the number of CD4(+) T cells. This is the first report demonstrating the actual amount of Vpr in HIV-1-positive patients, and the possible linkage of Vpr and viral titers indicates that it is important to continue to carry out the sequential analysis of Vpr, especially in clinical courses of HIV-1-positive patients. The threshold of viral titers, where Vpr appears in the patients' plasma, if present, contributes to better understanding the role of Vpr in AIDS pathogenesis. PMID- 17411373 TI - Leukemia inhibitor factor (LIF) inhibits HIV-1 replication via restriction of stat 3 activation. AB - Leukemia inhibitor factor (LIF) has been shown to potently inhibit HIV-1 replication in vitro and in human organ explant cultures. Furthermore, LIF activates the Jak/Stat signaling pathway with which many viruses, including HIV 1, interfere. We used CXCR4 and the LIF signaling receptor (gp130)-expressing cMAGI cells transfected with CD4, CCR5, and HIV-LTR-beta-galactosidase as a model system to investigate the potential involvement of Stat proteins in the anti-HIV 1 effect of LIF. Pretreatment with recombinant human (rh)LIF resulted in a significantly reduced uptake of HIV-1(BaL) , HIV-1(LAI), and SIVmac251 viral particles without affecting uptake of murine leukemia retroviral particles. HIV 1(BaL), HIV-1(LAI), as well as rhLIF selectively induced phosphorylation of Stat 3 but not Stat 1 or Stat 5. However, treatment of cMAGI cells with rhLIF prior to HIV-1 infection downregulated the HIV-1-mediated Stat 3 phosphorylation. In addition, peripheral blood mononuclear cells (PBMCs) transfected with Stat 3 siRNA prior to HIV-1(LAI) or HIV-1(BaL) infection produced significantly less HIV 1 p24 antigen as compared to nontransfected HIV-1(LAI) and HIV-1(BaL)-infected PBMCs. Thus, the Jak/Stat signaling pathway is important for the HIV-1 replication life cycle and rhLIF excerts its anti-HIV-1 activity by disrupting this signaling cascade. PMID- 17411375 TI - Structural basis for coreceptor selectivity by the HIV type 1 V3 loop. AB - The third variable region (V3) of the HIV-1 surface glycoprotein, gp120, plays a central role in the interaction of the virus envelope with the cell surface chemokine receptors, triggering membrane fusion and virus entry into human lymphocytes and macrophages. The CXCR4 and CCR5 chemokine receptors are used by "X4-tropic" and "R5-tropic" viruses, respectively. Recently, the crown of the V3 loop was shown to bear a close structural homology to the beta2-beta3 loop in the CXC and CC chemokines, the natural ligands of CXCR4 and CCR5, respectively. This homology can serve as the foundation for 3D molecular modeling of the V3 loops from primary isolates whose coreceptor usage was experimentally defined. The modeling revealed a charged "patch" on the surface of V3 that correlates with coreceptor usage. This V3 surface patch is positively charged in X4-tropic viruses and negatively charged or neutral in R5-tropic viruses, and is formed by two amino acids, at position 11 and at position 24 or 25; amino acids 11 and 24 or 11 and 25 contact each other in 3D space. Residues at positions 11 and 25 were known previously to influence coreceptor usage, and the charge of the residues at these two positions is often used to predict viral tropism. However, we found that the predictive value of using the charge of residues 11, 24, and 25 to identify X4 or R5 tropism was improved over using only the charge of residues 11 and 25. Thus, the data suggest a new " 11/24/25 rule" : a positively charged amino acid at position 11, 24, or 25 defines X4; otherwise R5. This rule gave an overall predictive value of 94% for 217 viruses whose tropism had been determined experimentally as either X4 or R5. The results have additional implications for the design of HIV therapeutics, vaccines, and strategies for monitoring disease progression. PMID- 17411374 TI - Genotypic resistance to antiretroviral drugs in patients infected with several HIV type 1 genetic forms in Cuba. AB - The main objective of this study is to evaluate the prevalence of resistance associated mutations to reverse transcriptase (RT) and protease (PR) inhibitors (I) 2 years after the introduction of antiretroviral treatment in Cuba, analyzing the mutations corresponding to different HIV-1 genetic forms circulating in Cuba. A total of 425 plasma samples were collected in 2003, corresponding to 175 (41.2%) subtype B and 250 (58.8%) non-B genetic forms, including 56 (22.4 %) non B subtypes, 112 (44.8%) circulating recombinant forms (CRFs), and 82 (32.8%) unique RFs (URFs). Of these, 175 (41.2%) patients were under highly active antiretroviral therapy (HAART) and 250 (58.8%) were treatment-naive. The presence of RT and PR resistance-associated mutations was established by sequencing. Levels of resistance were evaluated according to the Stanford Database program (http://hivdb.stanford.edu). The prevalence of resistance to RTI was 52.2% among RTI-treated patients, 51.5% for subtype B, and 53.2% for non-B genetic forms, including CRF18_cpx, CRF19_cpx, subtype C, and BG URF. In treatment-naive patients it was 6.4% in subtype B and 4.2% in non-B subtypes and RFs. The prevalence of resistance to PRI was 30% among PRI-treated patients, 28% in subtype B and 31% in non-B genetic forms, and 3.2% among treatment-naive subjects, mostly BG recombinants. In conclusion, significant differences in the prevalence of resistance to RTI and PRI were not detected among the most frequent genetic forms from treated patients, suggesting that the genetic diversity of HIV 1 in Cuba does not play a main role in the development of resistance to antiretroviral drugs. The presence of transmitted resistance mutations supports the study of resistance at baseline of treatment. PMID- 17411376 TI - HLA-DR-restricted peptides identified in the Nef protein can induce HIV type 1 specific IL-2/IFN-gamma-secreting CD4+ and CD4+ /CD8+ T cells in humans after lipopeptide vaccination. AB - We screened the Neflaiprotein to identify new HLA-DR-restricted epitopes, because this small protein is expressed early during infection, and specific CD4(+) T cells are critical for effective immunity in HIV-1 infection. We synthesized a set of peptides that covers the sequence of the Nef protein, and performed binding assays using 10 common HLA-DR molecules. We defined four large regions in this protein able to bind very efficiently to eight HLADR molecules. We took advantage of healthy volunteers immunized with an HIV-1 lipopeptide vaccine that contains three of the four HLA DR-restricted regions to investigate their capacities to stimulate T cells. In 11 vaccinated volunteers, typed for their class II molecules, we were able to correlate sequences of the vaccine displaying binding activities to specific HLA-DR molecules and the induction of CD4(+) T cell proliferation. To identify potential HLA-DR epitopes, we synthesized 31 15 mer peptides and showed that 26 bound to one or more HLA-DR molecules. Interestingly, 12 of the 26 15-mer peptides identified are included in the sequence of lipopeptides. We used IFN-gamma ELISPOT and flow cytometer assays to investigate the capacity of these potential CD4(+) T cell epitopes to induce specific T cell responses. We showed that seven of these peptides were able to stimulate HIV-specific T cell responses in five of six tested volunteers. These cells are Nef-specific CD4(+) and CD4(+) CD8(+) T cells secreting IL-2/INF-gamma or IL-2 alone. To conclude, these 26 Nef HLA-DR-restricted peptides could be helpful to better evaluate CD4(+) deficiencies in HIV infection and, for new vaccine designs. PMID- 17411377 TI - CD25+ regulatory T cells isolated from HIV-infected individuals suppress the cytolytic and nonlytic antiviral activity of HIV-specific CD8+ T cells in vitro. AB - HIV infection is characterized by CD4(+) T cell depletion and progressive immune dysfunction; particularly impacted are HIV-specific T cell responses. An important component of immune-mediated control of HIV replication, killing of infected cells, appears to be impaired, in part due to poor cytolytic activity of HIV-specific cytotoxic T cells (CTL). In vitro, several functions of HIV-specific T cells, such as cytokine production, can be enhanced by the depletion of the immunosuppressive CD25(+) FoxP3(+) CD4(+) regulatory (Treg) cell subset. However, the effect of CD25(+) Treg cells on virus-specific cytolytic activity in the context of HIV or any human viral infection has not been investigated. The present study demonstrates that CD25(+) Treg cells isolated from the peripheral blood of HIV-infected subjects significantly suppress HIV Gag-specific cytolytic activity in vitro. In addition, CD25(+) Treg cells suppress effector function (coexpression of TNF-alpha and IFN-gamma) of HIV-specific CD8(+) T cells that proliferate in response to HIV antigen. Finally, the secretion of HIV-inhibitory CC-chemokines by HIV-specific and nonspecific CD8(+) T cells is significantly reduced in the presence of CD25(+) Treg cells. These data suggest that CD25(+) Treg-mediated suppression of the antiviral activity of HIV-specific CD8(+) T cells could impact the ability of HIV-infected individuals to control HIV replication in vivo. PMID- 17411379 TI - Transduction with human telomerase reverse transcriptase immortalizes a rhesus macaque CD8+ T cell clone with maintenance of surface marker phenotype and function. AB - T cell lines and clones play a key role in basic studies of cellular immunology, and are also finding applications in adoptive immunotherapy. However, with proliferative expansion, T cells ultimately undergo cellular senescence and death, so that long-term culture of T cell clones is difficult to achieve. Expression of telomerase reverse transcriptase (TERT) in differentiated cells can maintain telomere length over many cell divisions, preventing senescence. We used a retroviral vector that expresses the human TERT (hTERT) gene to transduce a rhesus macaque-derived CD8(+) T cell clone specific for the MamuA*01-restricted immunodominant SIV gag epitope CM9. Extensive in vitro characterization revealed that the untransduced parental cells and the hTERT-transduced cells displayed comparable proliferation capacity, effector memory surface marker profiles, cytolytic activities, and cytokine profiles following antigen stimulation. The hTERT-transduced cells showed improved survival compared to parallel nontransduced cultures during in vitro propagation in long-term culture. Such immortalized T cells may be useful as a source of consistent controls for in vitro assays of cellular immune function, and as a potentially important reagent for autologous adoptive cellular immunotherapy studies in macaques. PMID- 17411378 TI - Lack of KIR3DS1 binding to MHC class I Bw4 tetramers in complex with CD8+ T cell epitopes. AB - In HIV-1 infection, the synergistic association of a subset of Bw4 MHC class I molecules and the activating killer inhibitory receptor (KIR), KIR3DS1, with prolonged AIDS-free survival has been reported. As KIRs represent a diverse group of MHC class I receptors, we questioned whether Bw4 MHC class I molecules expressing isoleucine at position 80 (Bw4Ile80) and in complex with HIV-1-derived T cell epitopes represented KIR3DS1 ligands. MHC class I tetramers are powerful tools for the detection of T cell receptor-MHC class I interactions, and have recently been used to evaluate KIR-MHC class I binding ex vivo. Specifically, this approach has been successfully utilized to assess binding of Bw4 MHC class I tetramers to KIR3DL1, an inhibitory KIR and allele of KIR3DS1. In this study we generated a diverse panel of HIV-1-specific Bw4Ile80 MHC class I tetramers and tested its ability to bind transiently expressed KIR3DS1 on 293-T cells. Using flow cytometry analysis, the expression of KIR3DS1 on 293-T cells was confirmed by anti-FLAG BioM2 staining, prior to incubation with PE-conjugated MHC class I tetramers. Despite choosing a broad array of peptide epitopes and diverse Bw4Ile80 MHC class I molecules, we were unable to detect tetramer binding to KIR3DS1. We speculate that our negative finding may be a consequence of the MHC class I molecules and peptide epitopes chosen, but could also relate to key amino acid differences that distinguish KIR3DS1 from KIR3DL1. PMID- 17411380 TI - Characterization of an HIV-1 group M variant that is distinct from the known subtypes. AB - We identified an HIV-1 variant that belongs to the M group, with limited similarity of short genetic regions (100-200 nt) to subtype K, but the remainder of the genome is unrelated to any established HIV-1 subtype. The isolate was obtained from an HIV-1-positive male, living in the Netherlands, who encountered the virus before 1989, most probably via heterosexual contact in Africa. We describe the full-length genome sequence of four biological clones that were obtained from two samples collected 5 years apart. At both time points all open reading frames were intact. Within the 5-year interval, the person received antiretroviral therapy with zalcitabine and zidovudine for almost 4 years. Evolution of drug-resistant variants is likely given the increase in viral RNA load to +/-10,000 copies/ml during the last year of treatment. Surprisingly, the only regular RT mutation acquired during this period was K70R, which suggests that the genetic background of this variant is perhaps not suitable for the generation of the standard 41L, 67N, and 215Y/F mutations that typically arise during prolonged, nonsuccessful, zidovudine treatment. Awaiting the discovery of at least two additional, epidemiologically unrelated patients with a phylogenetically related HIV-1 variant, we can designate this variant a new HIV-1 subtype, or a distinct branch of subtype K. PMID- 17411381 TI - HIV-1 envelope T cell epitope "hotspots " among mice and humans and among CD4+ and CD8+ T cell subpopulations. AB - HIV-1-specific T cell responses correlate with control of infection and disease, thus encouraging a full understanding of the peptides and antigen-processing mechanisms that govern T cell activation. We have previously demonstrated that CD4(+) T cell epitopes cluster nonrandomly within envelope protein "hotspot" regions. The current study was initiated to determine whether envelope-specific CD8(+) T cells might share epitope "hotspots" with the CD4(+) T cell population. Identification of CD8(+) T cell determinants by ELISPOT assays with peripheral blood mononuclear cells from four HIV-1-infected individuals, in conjunction with a survey of determinants in the Los Alamos database, revealed similarities among "hotspot" positions for CD4(+) and CD8T(+) cells within mice and humans. These results emphasized the important influence that envelope peptide position may have on antigen processing, and the consequent impact such processing may have on HIV-1-specific CD4(+) and CD8(+) T-cell activities. PMID- 17411382 TI - HIV type 1 subtype C gag and nef diversity in Southern Africa. AB - Several HIV-1 subtype C-specific gag- and/or nef-based vaccines are currently intended for clinical trial in southern Africa. Here we provide sequences of 64 gag and 45 nef genes sampled in Malawi, Zambia, Zimbabwe, and South Africa and assess the degree of southern African HIV-1 diversity that will confront these vaccines. Whereas reasonable phylogenetic evidence exists for geographical clustering of subtype C gag and nef sequences from various other parts of the world, there is little evidence of similar population founder effects in the southern African epidemic. The entire breadth of subtype C diversity is represented in the southern African genes suggesting there may be no advantage in producing region- or country-specific subtype C vaccines. We do not, however, find much evidence of intersubtype recombination in the Southern African genes, implying that the likelihood of vaccine failure due to the emergence of intersubtype recombinants is probably low. PMID- 17411383 TI - HIV type 1 drug resistance among naive patients from Venezuela. AB - In this study, we characterize proviral DNA of 20 HIV-1 asymptomatic antiretroviral-naive patients from Venezuela in env, gag, and pol genes regions. Results from both env/gag HMA subtyping and phylogenetic analysis of pol partial sequences led to the description of clade B in all cases. Nevertheless, the high prevalence of polymorphisms was particularly evident among the protease sequences. A 10% prevalence of major resistance mutations to RTIs was found. Our data also suggested that the protease polymorphisms I62T and V77T could be considered as molecular markers of the subtype B local epidemic. In addition, we show how proviral DNA can be used as a reliable tool to follow trends of resistance mutation transmission. PMID- 17411385 TI - Theory-based predictors of condom use among university students in the United States and South Africa. AB - This study compared the predictive value of the theory of planned behavior in university students in South Africa (N = 251) and the United States (N = 160) who completed an anonymous self-administered questionnaire. Multiple regression analyses revealed that condom use and intention were significantly predicted by positive condom attitude, subjective norm, self-efficacy, and attending university in South Africa. Significant interactions between country and predictors indicated that subjective norm predicted condom use and intention more strongly in the American sample than in the South African sample; attitude predicted intention more strongly in the American sample than in the South African sample; but self-efficacy predicted intention more strongly in the South African sample than in the American sample. The theory of planned behavior may provide a useful framework for interventions to reduce South African students' risk of HIV/and sexuality transmitted diseases. Such interventions should especially focus on building self-efficacy to use condoms. PMID- 17411386 TI - Lower injection-related HIV-1 risk associated with participation in a harm reduction program in Kazan, Russia. AB - This study was carried out to investigate the extent of unsafe practices associated with participation in a harm reduction program in Kazan, Russian Federation. In this cross-sectional study a convenience sample of active drug injectors encountered by the field outreach teams was interviewed. Demographic data and information on injection drug use were obtained using a structured survey instrument. Comparisons between program clients and newly encountered individuals not yet engaged by the program were made. Clients were divided into four groups based on the duration and intensity of their interaction with the program. Clients were found to be less likely than newly encountered injectors to give away or use previously used syringes, more likely to have used a new syringe the last time they injected with others, and more likely to be able to anticipate their need to acquire new syringes. However, the sharing of nonsyringe injection paraphernalia was no different between clients and newly encountered injectors. Despite the limitations of a cross-sectional study, it appears that engagement with the harm reduction program in Kazan was associated with reduction in many aspects of unsafe injecting. PMID- 17411387 TI - Responsibility for HIV prevention: patterns of attribution among HIV-seropositive gay and bisexual men. AB - The Seroconversion Narratives for AIDS Prevention (SNAP) study elicited narratives from recently infected seropositive gay and bisexual men that described the circumstances of their own seroconversion. This analysis of the narratives explored participants' attributions of responsibility for HIV prevention before and after they became infected. Before becoming infected with HIV, responsibility for prevention was often attributed to HIV-negative individuals themselves. These retrospective attributions revealed themes that included feelings of negligence, a sense of consequences, followed by regret. After seroconversion, responsibility for HIV prevention was primarily attributed to HIV-positive individuals themselves. Themes within these attributions included pledges to avoid HIV transmission, a strong sense of burden related to the possibility of infecting someone, and risk reduction strategies that they implemented in an attempt to avoid HIV transmission. Greater understanding of ideas related to responsibility has the potential to increase the effectiveness of HIV prevention interventions. PMID- 17411388 TI - Exposure to MTV's global HIV prevention campaign in Kathmandu, Nepal; Sao Paulo, Brazil; and Dakar, Senegal. AB - In 2002 MTV aired a global media campaign, "Staying Alive," to promote HIV prevention among 16- to 25-year-olds. Skeptics believed that a global MTV campaign would reach only a small group of elite young people. MTV increased access to its campaign, however, by making all materials "rights free" to third party (non-MTV) broadcasters. Over 789 million households in over 166 countries had access to some or all of the campaign. To understand the level of actual exposure and the types of young people exposed, data were analyzed from population-based household surveys in three diverse urban areas where a campaign evaluation was conducted: Kathmandu, Nepal; Sao Paulo, Brazil and Dakar, Senegal. Exposure rates ranged from 12% in Kathmandu, 23% in Sao Paulo, and 82% in Dakar, reaching an estimated 32,000, 400,000, 220,000 16- to 25-year-olds in each city, respectively. A number of personal, social and economic characteristics found to predict campaign exposure were identified in each site; in general, these were related to economic status and use of "new" media technologies. Though this skew toward more exposure by those with greater resources existed, we found that the campaign audience was in no way composed only of "elite" young people. (For example, although more of those exposed to the campaign had used the Internet compared with those not exposed, this was not the majority of those exposed in most countries.) The possibility of reaching millions of young people through global networks with minimal marginal costs after production, creates a new paradigm for reaching an important segment of young people. PMID- 17411389 TI - MTV's "Staying Alive" global campaign promoted interpersonal communication about HIV and positive beliefs about HIV prevention. AB - In 2002 MTV launched a global multicomponent HIV prevention campaign, "Staying Alive," reaching over 166 countries worldwide. An evaluation of this campaign focused on three diverse sites: Kathmandu, Nepal; Sao Paulo, Brazil; and Dakar, Senegal. Data were collected before and after campaign implementation through population-based household surveys. Using linear regression techniques, our evaluation examined the effects of campaign exposure on interpersonal communication about HIV and the effects of campaign exposure and interpersonal communication on beliefs about HIV prevention. We found a consistent positive effect of exposure on interpersonal communication across all sites, though there were differences among sites with regard to whom the respondent talked about HIV. We also found a consistent positive effect of exposure on HIV prevention beliefs across sites when interpersonal communication was simultaneously entered into the model. Finally, in two sites we found a relationship between interpersonal communication and HIV prevention beliefs, controlling for exposure, though again, the effects differed by the type of person the communication was with. These similar findings in three diverse sites provide ecological validity of the findings that "Staying Alive" promoted interpersonal communication and influenced young people's beliefs about HIV prevention in a positive way, evidence for the potential of a global media campaign to have an impact on social norms. PMID- 17411390 TI - Substance use and sexual behaviors among Japanese tourists, students, and temporary workers in Honolulu, Hawaii. AB - A total of 249 Japanese nationals - tourists (n = 107), students (n = 98), and temporary workers (n = 44) - were recruited at the targeted community venues in Honolulu, Hawaii, and completed a structured survey questionnaire. Reported lifetime sexually transmitted diseases, or STDs infection (10% male and 20% female participants), and HIV infection rates (7%, 2 out of 31 persons tested) were high. Male participants were more likely to practice safe sex with female sex workers than with steady and casual female partners both in Japan and Hawaii. More than 80% of the participants reported having had sex under the influence of alcohol. Multivariate analysis revealed that positive attitudes toward drug use and negative attitudes toward condom use were significantly correlated with the frequency of sex under the influence of drugs with steady partners in the past 12 months. Future HIV/STD prevention intervention programs must target Japanese youths who are planning to visit Hawaii or elsewhere abroad, as well as Japanese high-risk groups (e.g., temporary workers in Hawaii), and provide information about HIV/STD prevention in relation to substance use. PMID- 17411391 TI - Prototypical images in condom scripts among AIDS-bereaved adolescents. AB - Twenty-five HIV-negative late adolescents (13 women and 12 men) who had lost a parent to AIDS generated vignettes in which the characters were deciding whether to use a condom (condom scripts). Two clinically trained judges rated the interpersonal tone of the condom scripts on 17 semantic differential scales. Three other clinically trained raters described script characters' attributes by selecting from a list of 36 terms. Multidimensional scaling (MDS) and individual differences hierarchical classes analyses (INDCLAS) were used to inductively derive a typology of condom scripts. Two dimensions emerged from MDS analysis: incompatibility and inequality. Condom scripts culminating in unprotected sex depicted situations in which partners held unequal influence. INDCLAS results suggested a prototype for equal-influence condom scripts - excited male and assertive, powerful female - and for unequal-influence (unprotected sex) condom scripts - powerful, disengaged male and permissive female. These results inform the development of theoretical models and HIV prevention program materials. PMID- 17411392 TI - Microarray data quality - review of current developments. AB - DNA microarray technologies have evolved rapidly to become a key high-throughput technology for the simultaneous measurement of the relative expression levels of thousands of individual genes. However, despite the widespread adoption of DNA microarray technology, there remains considerable uncertainty and scepticism regarding data obtained using these technologies. Comparing results from seemingly identical experiments from different laboratories or even from different days can prove challenging; these challenges increase further when data from different array platforms need to be compared. To comply with emerging regulations, the quality of the data generated from array experiments needs to be clearly demonstrated. This review describes several initiatives that aim to improve confidence in data generated by array experiments, including initiatives to develop standards for data reporting and storage, external spike-in controls, quality control procedures, best practice guidelines, and quality metrics. PMID- 17411393 TI - Self-self hybridization as an alternative experiment design to dye swap for two color microarrays. AB - Dye-specific bias effects, commonly observed in the two-color microarray platform, are normally corrected using the dye swap design. This design, however, is relatively expensive and labor-intensive. We propose a self-self hybridization design as an alternative to the dye swap design. In this design, the treated and control samples are labeled with Cy5 and Cy3 (or Cy3 and Cy5), respectively, without dye swap, along with a set of self-self hybridizations on the control sample. We compare this design with the dye swap design through investigation of mouse primary hepatocytes treated with three peroxisome proliferator-activated receptor-alpha (PPARalpha) agonists at three dose levels. Using Agilent's Whole Mouse Genome microarray, differentially expressed genes (DEG) were determined for both the self-self hybridization and dye swap designs. The DEG concordance between the two designs was over 80% across each dose treatment and chemical. Furthermore, 90% of DEG-associated biological pathways were in common between the designs, indicating that biological interpretations would be consistent. The reduced labor and expense for the self-self hybridization design make it an efficient substitute for the dye swap design. For example, in larger toxicogenomic studies, only about half the chips are required for the self-self hybridization design compared to that needed in the dye swap design. PMID- 17411394 TI - Application of DNA microarrays in the study of human obesity and type 2 diabetes. AB - DNA microarrays have provided medical researchers with a powerful tool to study the mechanisms of complex diseases, including obesity and type 2 diabetes (T2D). The technology has been used to dissect virtually every aspect of the genetic and molecular basis of these two diseases. Gene expression profiling is the major application of DNA microarrays so far. Subcutaneous fat, visceral fat, adipocyte and preadipocyte, muscle, liver, pancreas and specific nuclei in the hypothalamus under normal and disease conditions are used in addressing the profile of gene expression in obesity and T2D. Comparisons of fat depots in humans and animal models - including ob/ob and db/db mice, diet-induced obese mice, fa/fa Zucker rats, gene knockout (plin (-/-), GLUT4 (-/-)) and transgenic mice (GLUT4-Tg) - have been employed in microarray experiments. The effects of various interventions, such as hormonal and drug treatments, exercise, and surgery, have been studied to determine the expression profile of different developmental stages in cells and the effect of treatment on the two diseases. In this review, the application of microarrays in elucidating the role of retinol binding protein 4 as a link between obesity and T2D is discussed. The possible role in obesity of a common genetic variant near the INSIG2 gene and the discovery of the BBS9 gene are also discussed. The problems and challenges are summarized under eight categories and suggestions for the future direction of research in this area are proposed. PMID- 17411395 TI - Identification of functional cell adhesion molecules with a potential role in metastasis by a combination of in vivo phage display and in silico analysis. AB - Organ-specific homing of malignant cells involves interactions mediated through cell adhesion molecules and their receptors on the cell surface. Identification of peptides that mimic these receptor-ligand interactions is critical for analyzing the functional role of these proteins and is therapeutically significant to target or block organ-specific homing of tumor cells. Following three cycles of in vivo biopanning using a phage display peptide library injected into mice, we identified 11 unique peptides that were specific for homing to lung, liver, bone marrow, or brain. We developed a bioinformatics strategy to identify putative cell adhesion molecules (CAM) involved in tumor cell migration, invasion, and metastasis based on identified organ-specific peptides. Structural information, including surface exposure and the binding preference of any of these residues in the identified proteins, was examined. These studies resulted in identification of Semaphorin 5A (mouse, Sema5A; human, SEMA5A) and its receptor Plexin B3. The gene expression profile of these proteins in tumors and tumor cell lines was assessed using virtual microarray and serial analysis of gene expression (SAGE) databases and was further confirmed using reverse transcriptase polymerase chain reaction (RT-PCR). Our data demonstrate an association between the expression of SEMA5A and Plexin B3 and the aggressiveness of pancreatic and prostate cancer cells. In summary, using a combined experimental and bioinformatics approach, we have identified functional tumor specific CAMs, which may be critical for organ-specific metastasis. PMID- 17411396 TI - Comparative genomics of small RNAs in bacterial genomes. AB - In recent years, various families of small non-coding RNAs (sRNAs) have been discovered by experimental and computational approaches, both in bacterial and eukaryotic genomes. Although most of them await elucidation of their function, it has been reported that some play important roles in gene regulation. Here we carried out comparative genomics analysis of possible sRNAs that are computationally identified in 30 bacterial genomes from gamma- and alpha proteobacteria and Deinococcus radiodurans. Identified sRNAs are clustered by a complete-linkage clustering method to see conservation among the organisms. On average, sRNAs are found in approximately 30% of intergenic regions of each genome sequence. Of these, 25.7% are conserved among three or more organisms. Approximately 60% of the conserved sRNAs do not locate in orthologous intergenic regions, implying that sRNAs may be shuffled their positions in genomes. The current study implies that sRNAs may be involved in a more extensive range of functions in bacteria. PMID- 17411397 TI - Small RNA identification in Enterobacteriaceae using synteny and genomic backbone retention. AB - Genomic screens for small RNA candidates in Enterobacteriacae genomes were carried out with existing small RNA sequences, conserved flanking genes, and genomic backbone information. The small RNA sequences and contexts from E. coli K12 formed the basis of the search. Sequence identity identified 117 additional small RNA homologs in related genomes. Motifs of continuous sequence stretches added another 48 sRNA regions, termed partial homologs. However, this study is unique in identifying 160 nonhomologous sRNA loci in related genomes based on the conserved flanking gene synteny and the backbone retention information obtained from KEGG-SSDB. Gene synteny and genomic backbone continuity were observed to be correlated with all of the sRNAs in related genomes. This search is the first of its kind toward identification of functionally important regions using gene order and back-bone information. A disruption in flanking gene order or genomic backbone indicates a possible hotspot for alien gene pool integration. This study reports both occurrence of multiple copies of a sRNA and co-occurrence of different sRNAs between a pair of conserved flanking genes. In general, synteny and genomic backbone retention information can be added as additional search criteria toward the design of precise bioinformatics tools for sRNA, gene identification, and gene functional annotations in related genomes. PMID- 17411398 TI - Evolutionary-conserved gene expression response profiles across mammalian tissues. AB - Gene expression responses are complex and frequently involve the actions of many genes to effect coordinated patterns. We hypothesized these coordinated responses are evolutionarily conserved and used a comparison of human and mouse gene expression profiles to identify the most prominent conserved features across a set of normal mammalian tissues. Based on data from multiple studies across multiple tissues in human and mouse, 13 gene expression modes across multiple tissues were identified in each of these species using principal component analysis. Strikingly, 1-to-1 pairing of human and mouse modes was observed in 12 out of 13 modes obtained from the two species independently. These paired modes define evolutionarily conserved gene expression response modes (CGEMs). Notably, in this study we were able to extract biological responses that are not overwhelmed by laboratory-to-laboratory or species-to-species variation. Of the variation in our gene expression dataset, 84% can be explained using these CGEMs. Functional annotation was performed using Gene Ontology, pathway, and transcription factor binding site over representation. Our conclusion is that we found an unbiased way of obtaining conserved gene response modes that accounts for a considerable portion of gene expression variation in a given dataset, as well as validates the conservation of major gene expression response modes across the mammals. PMID- 17411399 TI - Consensus framework for exploring microarray data using multiple clustering methods. AB - The large variety of clustering algorithms and their variants can be daunting to researchers wishing to explore patterns within their microarray datasets. Furthermore, each clustering method has distinct biases in finding patterns within the data, and clusterings may not be reproducible across different algorithms. A consensus approach utilizing multiple algorithms can show where the various methods agree and expose robust patterns within the data. In this paper, we present a software package - Consense, written for R/Bioconductor - that utilizes such an approach to explore microarray datasets. Consense produces clustering results for each of the clustering methods and produces a report of metrics comparing the individual clusterings. A feature of Consense is identification of genes that cluster consistently with an index gene across methods. Utilizing simulated microarray data, sensitivity of the metrics to the biases of the different clustering algorithms is explored. The framework is easily extensible, allowing this tool to be used by other functional genomic data types, as well as other high-throughput OMICS data types generated from metabolomic and proteomic experiments. It also provides a flexible environment to benchmark new clustering algorithms. Consense is currently available as an installable R/Bioconductor package (http://www.ohsucancer.com/isrdev/consense/). PMID- 17411400 TI - The interface between the device and the patient: masks, mouthpieces, and "others." Introduction. PMID- 17411401 TI - Clinical side effects during aerosol therapy: cutaneous and ocular effects. AB - Aerosolized medications maximize clinical benefit by targeting the airways and minimize side effects by reducing (though not eliminating) systemic exposure. Aerosolized drugs delivered with a facemask may inadvertently deposit on the face and in the eyes, raising concerns about cutaneous and ocular side effects with these drugs. Cases of anisocoria have been reported from exposure of the eyes to aerosol bronchodilators. Whether inhaled corticosteroids (ICS) can cause skin and eye problems like those seen with systemic or topical steroids is more difficult to answer. Patients who take ICS may have other corticosteroid exposures, or have other conditions that predispose them to side effects, making the analysis of the ICS risk challenging. Also, many studies were not designed to search for cutaneous or ocular effects, or may have been too short to detect these effects. Nevertheless, ICS have been associated with an increased risk of skin thinning, bruising, cataracts and possibly glaucoma in adults, but not in children. The risks increase with advanced age, higher doses, and longer duration of use. In children, the risks of cataracts and glaucoma were negligible with ICS, whether a mouthpiece or a mask interface was used. Side effects like skin rash and conjunctivitis occurred at low frequencies similar to placebo or comparator drugs. We do not know whether exposed children will have increased risks from ICS later in life. Therefore, it is wise to avoid face and eye deposition when possible, and to use the minimally effective dose. PMID- 17411402 TI - Modeling of aerosol deposition with interface devices. AB - Various approaches can be used to mathematically model the performance of different masks, mouthpieces, and aerosol delivery devices. The sophistication of such models can vary widely, from the use of simple algebraic empirical correlations to advanced computational fluid dynamics simulations. Bench-top testing is also often used to model aspects of devices, since it is difficult to capture certain aspects of device behavior with mathematical models. These various approaches to modeling differ in their limitations. Empirical correlations exist for predicting the effects of varying mouthpiece diameter and mouth-throat dimensions on extrathoracic losses, but are restricted to stable, nonballistic aerosols in certain flow rate ranges. Computational fluid dynamics (CFD) simulations that solve the Reynolds-averaged Navier-Stokes (RANS) equations typically require near-wall turbulence corrections in order to adequately model mouth-throat deposition, while Large Eddy Simulation (LES) removes this deficiency. Bench-top models that use replicas of the extrathoracic airways vary in their accuracy and generality in replicating the filtering properties of these airways. Choosing and using these various modeling approaches for evaluating patient-device interfaces requires knowledge of their merits and pitfalls, a brief discussion of which is given here. PMID- 17411403 TI - Design of facemasks for delivery of aerosol-based medication via pressurized metered dose inhaler with valved holding chamber: key issues that affect performance. AB - Valved holding chambers (VHCs) are widely prescribed for use with pressurized metered dose inhalers (pMDIs) for the treatment of respiratory disease by aerosol therapy. The facemask is the preferred patient interface for use by infants and small children, as well as by geriatric patients, due primarily to poor coordination skills. However, care is required in the design of the facemask-VHC system to optimize the delivery of medication. In particular, it is essential to achieve an effective mask-to-face seal and to minimize the volume of dead space. It is also important to ensure that the fit of the facemask is comfortable to the patient when applied with sufficient force to create a seal. We review each of these design principles and their application in the evolution of a range of VHCs from the same family of devices during the past fifteen years. We also examine the various methods available for evaluating VHC-facemasks as a system, recommending where future work might be directed. PMID- 17411405 TI - Jet nebulizers versus pressurized metered dose inhalers with valved holding chambers: effects of the facemask on aerosol delivery. AB - The delivery of an aerosolized drug to a child is a complex process requiring an interaction between parent, child, and inhalation device. Recent studies have shown that the facemask can be a key factor affecting aerosol delivery, particularly the influence of leaks between the facemask and the face. To further quantify these effects and design around them, we have developed a bench model consisting of a breathing simulator, an inhaled mass filter, and a "pediatric face." This paper reviews the development of this model and details important decisions made in its configuration, particularly inhaled mass filter location (e.g., between device and facemask, or in mouth) and mouth diameter (4 or 18 mm). With the final design, we used the model to measure the impact of the "blow-by" technique on nebulizer inhaled mass. In a separate series of experiments, we studied the effects of a "crying" pediatric breathing pattern on inhaled mass for both nebulizers and pressurized metered dose inhalers with valved holding chambers (pMDI VHCs). Results indicated that the location of the inhaled mass filter was a critical factor in assessing aerosol delivery through facemasks and that the "mouth diameter" was not an important variable. Failure to locate the filter in the mouth behind the face, especially for jet nebulizers, failed to accurately measure effects of the facemask and significantly overestimated aerosol delivery. Blow-by results indicated that a 1-cm gap between the facemask and the face was not critical when using a front-loaded facemask. Finally, even with optimal design, the combination of an aerosol generator and facemask with a crying breathing pattern reduced the inhaled mass to 1% of the label dose. PMID- 17411406 TI - Devices for aerosol delivery to treat sinusitis. AB - Compared to research into aerosolized delivery of drugs to treat lung disease, research into nasal delivery of aerosolized drugs to treat sinusitis has been significantly neglected. This is despite the fact that more individuals suffer from sinusitis in the United States than suffer from asthma, and its consequences lead to considerable amounts of discomfort, lost work days, and money spent on health care. A number of studies have shown that a high proportion of aerosolized medications delivered by metered dose inhalers (MDIs) and aqueous spray devices deposits in the anterior one-third of the nasal cavity. However, the important targets for treating sinusitis lie beyond this region. These include the middle meatus, the superior and posterior regions of the nasal cavity and the sinuses themselves. This review examines the particle-related and device-related factors that are known to improve intranasal delivery of aerosolized medications to these targets and their efficacy in patients with disease. Based on this review, it is recommended that companies that are interested in improving aerosol delivery to treat sinusitis utilize both in vivo imaging modalities and in vitro models of the nasal cavity and sinuses to assess intranasal aerosol delivery and device performance during the development stage. Once device design has been optimized, it is recommended that device manufacturers and pharmaceutical companies move beyond the current reliance on anecdotal reporting and uncontrolled studies to clinical trials that are randomized and placebo-controlled and that quantify changes both in symptoms and in functional parameters to determine drug efficacy with their device. PMID- 17411407 TI - Facemasks and aerosol delivery by metered dose inhaler-valved holding chamber in young children: a tight seal makes the difference. AB - A facemask on a valved holding chamber (VHC) facilitates the inhalation of aerosols from metered dose inhalers (MDI) for young children. Only recently the facemask has been recognized as a vital part for efficient aerosol delivery. Several in vitro and in vivo studies show that a tight seal of the facemask is crucial for optimal aerosol deposition to the lungs. Even a small leak can reduce the dose delivered to the lungs considerably. However, a tight seal is difficult to obtain when a child is not cooperative. Depending on the design of the facemask, it is easier to obtain a good seal. Factors such as dead space, shape, and material should be considered when designing a facemask. However, when a child is upset and not cooperative during the administration, aerosol deposition will be minimal, even with the best-designed facemask. PMID- 17411408 TI - Facemask design, facial deposition, and delivered dose of nebulized aerosols. AB - Nebulizers are often interfaced to patients using facemasks, especially when the patient is sick and uncooperative. Tight-fitting masks are thought to improve drug delivery, but recent studies have indicated that facemask seal can impact facial and eye deposition of aerosol. The purpose of the present study was to define the factors that influence drug delivery to the lung in pediatric patients using nebulizers and facemasks; particularly the roles of facemask seal, mask vents and nebulizer flow. Using a pediatric face facsimile and radiolabeled saline aerosols front-loaded and bottom-loaded nebulizers were tested for aerosol delivery during a pediatric pattern of breathing. Gamma scintigraphy provided images of the face. Filters measured drug delivery to the patient (inhaled mass [IM]). All data were reported as percent (%) nebulizer charge. Nebulizer flows of 4 and 8 L/min were tested. Preliminary experiments suggested that inertial forces between the edge of the mask and the face were responsible for facial and eye deposition. Front-loaded nebulizers were more efficient than bottom-loaded systems in delivering drug to the patient but favored eye deposition. These observations led to the design of a mask prototype constructed to maximize aerosol delivery to the patient with reduced deposition on the face and in the eyes. Modifications included vents and specialized cutouts in the region of the eyes. A tight fitting front-loaded mask delivered an IM of 6.38 +/- 0.42% (mean +/- SE) with facial and eye deposition of 1.76 +/- 0.17% and 1.14 +/- 0.15% respectively. The presence of specialized eye cutouts minimized facial and eye deposition (0.72 +/- 0.07%, and 0.15 +/- 0.02% [P < 0.0001]), even in the presence of increased nebulizer flow. The prototype design at 4 L/min maximized IM to 8.78 +/- 0.98% and further reduced facial and eye deposition (0.66 +/- 0.07% and 0.09 +/- 0.01%). Commercial bottom loaded masks reduced IM to 2.33 +/- 0.22%, with significant deposition on the face (1.43 +/- 0.16%). For aerosol therapy with nebulizers in pediatric patients, facemask design is a key factor in maximizing aerosol delivery to the patient while minimizing deposition on the face and in the eyes. PMID- 17411409 TI - Facemasks and aerosol delivery in vivo. AB - It has been shown in vitro that even a small air leak in the facemask can drastically reduce the efficiency of drug delivery. In addition, it has been shown that drug deposition on the face does significantly add to overall drug loss and has the potential of local side effects. The aim of this study is therefore to verify these findings in vivo. Eight asymptomatic recurrently wheezy children, aged 18-36 months, inhaled a radiolabeled salbutamol formulation either from a pressurized metered-dose inhaler through a spacer with attached facemask or from a nebulizer with attached facemask. Drug deposition of radiolabeled salbutamol was assessed with a gamma camera and expressed as a percentage of the total dose. Lung deposition expressed as a percentage of the total dose (metered dose and nebulizer fill, respectively) was 0.2% and 0.3% in children who inhaled with a non-tightly fitted facemask. Lung deposition was 0.6% and 1.4% in screaming children with a tightly fitted facemask and between 4.8% and 8.2% in patients breathing normally. Overall mask deposition was between 0.8% and 5.2%. Overall face deposition was between 2.6% and 8.4%. The results from this pilot study support the results found in in vitro studies, where a facemask leak greatly reduces drug delivery to the patient. PMID- 17411410 TI - The mask for noninvasive ventilation: principles of design and effects on aerosol delivery. AB - There has been much clinical and academic interest in the use of noninvasive positive-pressure ventilation (NPPV) in patients with acute and chronic respiratory failure. The use of NPPV in appropriately selected patients improves survival and decreases the need for endotracheal intubation. The most commonly used interfaces for NPPV are nasal masks or oronasal masks, but nasal pillows, mouthpieces, total face masks, and helmets can also be used. Critical care ventilators, portable volume ventilators, and ventilators designed specifically for NPPV can be used. There are three options for aerosol delivery during NPPV. The patient can be removed from NPPV and the aerosol administered by nebulizer or MDI in the standard manner, the aerosol can be delivered by nebulizer placed in line between the circuit and the mask, or a spacer chamber can be placed between the circuit and the mask. There is presently no commercially available system designed specifically for aerosol delivery during NPPV with a bilevel (BiPAP) ventilator. However, in vitro and in vivo studies have demonstrated that a significant amount of bronchodilator can be administered by in-line nebulizer or MDI during NPPV. The evidence base for aerosol delivery during NPPV is not nearly as mature as the evidence for aerosol delivery during invasive mechanical ventilation. With NPVV, issues related to the optimal interface, ventilator settings, and aerosol generator (nebulizer versus MDI) are largely unexplored. PMID- 17411412 TI - Therapeutic effect of pEgr-IL18-B7.2 gene radiotherapy in B16 melanoma-bearing mice. AB - To evaluate the antitumor role of genes B7.2 and IL18, the radiation-inducible dual-gene coexpression plasmid pEgr-IL18-B7.2 was constructed and its effects on tumor were detected both in vitro and in vivo. After the introduction of pEgr IL18-B7.2 into B16 melanoma cells, followed by X-ray irradiation, higher expression levels of B7.2 and IL18 compared with control were found both by flow cytometry and enzyme-linked immunosorbent assay. It was shown that even low-dose irradiation was able to induce their expression, which could be tightly regulated either by giving cells different doses of radiation or the same dose at different time points. pEgr-IL18-B7.2 was then packaged with liposome and injected into melanoma tumor-bearing mice. The tumors received 5 Gy of local X-ray irradiation every other day for a total of five treatments. B16 tumor growth slowed significantly when treated with pEgr-IL18-B7.2 plus X-radiation versus either treatment separately. Both 1 and 3 days after the last irradiation the group of mice with combined gene and radiation therapy showed significantly higher tumor necrosis factor (TNF)-alpha secretion in peritoneal macrophages, upregulated splenic cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells, and higher interferon (IFN)-gamma secretion than those in either individual treatment group or the control group. The stimulation of host anticancer immunity by increased secretion of IL-18 and upregulated immunogenicity of the tumor cells by increased expression of B7.2 on their surface, in addition to the direct effect of local X irradiation on the tumor cells, may contribute to the novel effect of the combined therapy. PMID- 17411413 TI - Hospital- versus community-based syringe exchange: a randomized controlled trial. AB - This study examined the effect of syringe exchange program setting on the injection practices, health status, and health service utilization patterns of injection drug users (IDUs) recruited from a public urban hospital. One hundred sixty-six participants were randomized to either community- or hospital-based syringe exchange services. Poisson regression models were used to compare service utilization between groups. In both conditions, risky drug use practices decreased, and physical health functioning improved over time. Hospital-based syringe exchange program (SEP) attendees had 83% more inpatient admissions (p < .0001) and 22% more ambulatory care visits (p < .0001) than those assigned to the community-based SEP condition. Syringe exchange services that are integrated into public hospital settings may serve as a valuable strategy to engage hard to reach IDU populations in behavioral interventions designed to reduce HIV risk transmission behaviors and increase access to, or engagement in, the use of secondary and tertiary preventive medical care. PMID- 17411414 TI - Residual injection risk behavior, HIV infection, and the evaluation of syringe exchange programs. AB - This study assessed relationships between residual risk behavior (risk behavior among persons participating in effective HIV prevention programs) and HIV infection. Structured interviews and HIV tests were obtained from participants in six large U.S. syringe exchange programs. Program characteristics were obtained through interviews with the directors. Findings indicated that injection risk behaviors varied significantly across the six programs--from 10% to 27% of the participants at each program reported receptive sharing of needles and syringes in the 30 days prior to the interview. HIV prevalence ranged from 2.5% to 22.2% across the six programs. HIV prevalence among new injectors was strongly related to HIV prevalence among long-term injectors across the programs (r = .869). There was a consistent pattern of negative relationships between injection risk behaviors and HIV infection across the six programs (higher rates of risk behavior at a program associated with lower HIV infection). As a result, appropriate evaluation of HIV prevention programs may require not only information on continuing risk behavior and HIV infection among program participants but also historical information on the epidemiology of HIV in the local community. PMID- 17411415 TI - Reduction in injection-related HIV risk after 6 months in a low-threshold methadone treatment program. AB - This study assessed injection-related HIV risk behavioral changes among opioid users 6 months after enrollment in low-threshold (harm reduction based) metha done maintenance treatment (MMT) programs within needle exchange services in Kingston and Toronto, Ontario, Canada. Changes were assessed for all participants (whole cohort), participants who continued to use illicit drugs by any route (drug-using subcohort); and those who continued to inject drugs (injecting subcohort). In this prospective observational cohort study, an interviewer administered questionnaire examining injection-related HIV risk behaviors was administered to 183 study participants at entry to treatment and 6 months later. Changes in risk behaviors were analyzed using conditional logistic regression which took into account the paired nature of the data. We found that the proportion of participants injecting drugs, sharing needles, sharing drug equipment, indirectly sharing and using shooting galleries declined with follow up for the whole cohort. Within the drug-using subcohort, there was a decrease in the proportion of individuals who injected drugs, while within the injecting subcohort the sharing of injection equipment and the use of shooting galleries declined. Our findings suggest that low-threshold MMT programs can reduce the risk of HIV without the enforcement of abstinence-based policies. PMID- 17411416 TI - HIV risk behaviors among rural stimulant users: variation by gender and race/ethnicity. AB - We examined data from a community sample of rural stimulant users (n = 691) in three diverse states to identify gender and racial/ethnic differences in HIV risk behaviors. Bivariate and logistic regression analyses were conducted with six risk behaviors as dependent variables: injecting drugs, trading sex to obtain money or drugs, trading money or drugs to obtain sex, inconsistent condom use, multiple sex partners, and using drugs with sex. Controlling for state, income, age, heavy drinking, and type of stimulant used, men had lower odds than women for trading sex to obtain money or drugs (adjusted odds ratio [AOR] =0.4, confidence interval [CI] = 0.28-0.59; p < .0001), greater odds than women for trading money or drugs to obtain sex (AOR = 44.4, CI = 20.30-97.09; p < .0001), greater odds than women of injecting drugs (adjusted odds ratio (AOR =1.6, CI = 1.11-2.42; p = .01), and lower odds than women of using condoms inconsistently (AOR = 0.6, CI = 0.35-0.92; p = .02); African Americans had lower odds than Whites of injecting drugs (AOR = .08, CI = 0.04-0.16; p < .0001), greater odds than Whites for trading sex to obtain money or drugs (AOR = 1.7, CI = 1.01-2.85; p = .04) and for trading money or drugs to obtain sex (AOR = 2.9, CI = 1.53-5.59; p = .001), and greater odds than Whites of using drugs with sex (AOR = 3.9, CI = 1.47-10.09; p = .006). These findings indicate HIV prevention efforts should be tailored to address gender and racial/ethnic differences in risk behaviors among rural stimulant users. PMID- 17411417 TI - Acquisition of new sexual partners among women with HIV infection: patterns of disclosure and sexual behavior within new partnerships. AB - This study describes the sexual behavior of HIV-positive women within new versus more established relationships and determines whether beliefs about HIV antiretroviral therapy (ART) impact these behaviors. The Women's Interagency HIV Study is a longitudinal cohort study of HIV among women in the United States. Sexually active HIV-positive women (N = 1,090) completed interviews on beliefs and behaviors at 6-month intervals. Data were analyzed for the period between April 2002 and March 2003. Of 1,517 sexual partners reported, 32% were newly acquired within the previous 6 months. As compared with more established sexual relationships, newer partnerships were characterized by greater condom use consistency (odds ratio = 1.8, 95% confidence interval = 1.4 -2.3). Holding beliefs that ART is protective for HIV transmission impacted the relationship between partner type and condom use. In established relationships, 63% reported consistent condom use if they believed that ART is not protective, whereas 54% reported consistent condom use if they believed that ART is protective. CONCLUSIONS: These findings highlight the importance of ongoing support for sexual risk reduction among women with HIV-infection and for strategies that reduce the strength of relationships between ART beliefs and sexual risk behavior. PMID- 17411418 TI - Social and behavioral determinants of consistent condom use among female commercial sex workers in Ghana. AB - This study investigated the social and behavioral predictors of consistent condom use among female commercial sex workers (FCSWs) in Ghana. Four hundred fifty street commercial sex workers were interviewed in Accra, Kumasi and Techiman. The level of condom education was very low (14%); however consistent condom use (all the time) with clients was relatively high (49.6%). Two hundred seventy-seven of the participants did not use condoms all the time. PMID- 17411419 TI - Key components of a theory-guided HIV prevention outreach model: pre-outreach preparation, community assessment, and a network of key informants. AB - Although outreach frequently is used to provide community-based HIV prevention services to members of underserved populations, researchers may not be familiar with the specific components of and factors influencing outreach and how systematic community outreach methods can be used to recruit participants for research purposes. This article describes key components of a theory and PRECEDE based outreach model developed and used as part of a broader study examining the feasibility of enhancing access to STD clinical services for sexually transmitted diseases in order to reduce HIV incidence. We present a three-part outreach model and describe lessons learned from implementing it. Factors that improved access to key informants who could facilitate participant recruitment during the outreach process included sustained project visibility in the community, outreach worker affiliation with trusted community-based organizations, and development of a reliable network of key informants. This model enabled a systematic approach to reaching community members and documenting the steps taken to do so. PMID- 17411420 TI - The peptide-binding activity of GRP94 is regulated by calcium. AB - GRP94 (glucose-regulated protein of 94 kDa) is a major luminal constituent of the endoplasmic reticulum with known high capacity for calcium in vivo and a peptide binding activity in vitro. In the present study, we show that Ca2+ regulates the ability of GRP94 to bind peptides. This effect is due to a Ca2+-binding site located in the charged linker domain of GRP94, which, when occupied, enhances the association of peptides with the peptide-binding site in the N-terminal domain of the protein. We further show that grp94-/- cells are hypersensitive to perturbation of intracellular calcium and thus GRP94 is important for cellular Ca2+ storage. PMID- 17411421 TI - Civil conflict and sleeping sickness in Africa in general and Uganda in particular. AB - Conflict and war have long been recognized as determinants of infectious disease risk. Re-emergence of epidemic sleeping sickness in sub-Saharan Africa since the 1970s has coincided with extensive civil conflict in affected regions. Sleeping sickness incidence has placed increasing pressure on the health resources of countries already burdened by malaria, HIV/AIDS, and tuberculosis. In areas of Sudan, the Democratic Republic of the Congo, and Angola, sleeping sickness occurs in epidemic proportions, and is the first or second greatest cause of mortality in some areas, ahead of HIV/AIDS. In Uganda, there is evidence of increasing spread and establishment of new foci in central districts. Conflict is an important determinant of sleeping sickness outbreaks, and has contributed to disease resurgence. This paper presents a review and characterization of the processes by which conflict has contributed to the occurrence of sleeping sickness in Africa. Conflict contributes to disease risk by affecting the transmission potential of sleeping sickness via economic impacts, degradation of health systems and services, internal displacement of populations, regional insecurity, and reduced access for humanitarian support. Particular focus is given to the case of sleeping sickness in south-eastern Uganda, where incidence increase is expected to continue. Disease intervention is constrained in regions with high insecurity; in these areas, political stabilization, localized deployment of health resources, increased administrative integration and national capacity are required to mitigate incidence. Conflict-related variables should be explicitly integrated into risk mapping and prioritization of targeted sleeping sickness research and mitigation initiatives. PMID- 17411422 TI - Presumed stromal graft rejection after automated lamellar therapeutic keratoplasty: case report. AB - PURPOSE: To describe the development of presumed immune-mediated stromal rejection after automated lamellar therapeutic keratoplasty (ALTK) and its reversal after initiation of intensive topical corticosteroid therapy. METHODS: Observational case report. RESULTS: Stromal edema localized in the graft developed 42 days after ALTK for Avellino corneal dystrophy in a 65-year-old man. After one week of intensive topical corticosteroids, complete reversal of graft edema occurred, with full recovery of visual function. CONCLUSION: The clinical appearance and response to therapy in this case supported the diagnosis of immune mediated stromal rejection. Ophthalmologists should be aware that stromal rejection may occur in lamellar corneal grafts. PMID- 17411423 TI - Drama-based education to motivate participation in substance abuse prevention. AB - BACKGROUND: The substance abuse prevention goal of the theatre production "TUNNELS" was to provide community education on substance abuse to an audience in Durham, NC and surrounding communities. The education effort intended to increase awareness and understanding of the risk and protective factors associated with alcohol and other drug use, and to promote pro-active behaviors in substance abuse prevention within the adult community. It was hypothesized that community based education via drama would change attitudes toward alcohol and substance abuse, and increase participation in family and community activities aimed at substance abuse prevention. METHODS: A focus group comprised of educators, substance abuse researchers and local substance abuse counselors developed "life stories" of users of alcohol and other drugs and a local playwright incorporated these and other experiences into a series of six vignettes. The production was publicized throughout the Durham area, and 700 adults attending the play signed a consent form and completed the pre-play survey. The participant pool was restricted to those adults who completed both the time-1 and time-2 surveys and resided within Durham and surrounding communities. Paired comparisons of mean responses were analyzed using a paired sample two-tailed t-test. A telephone survey three months after the play assessed attitudes toward substance abuse as a disease, and whether the respondents had increased their participation in prevention activities including discussions of the play with others. RESULTS: Viewing the play increased the knowledge base of participants regarding substance abuse as a disease, even though the audience demonstrated an appreciation of risk and protective factors prior to attending the performance. In the pre-play survey, participants indicated a strong opinion that parental involvement in teen life was important, and therefore this was not increased as a result of viewing the play. It was found that the drama increased intent to participate in substance abuse prevention activities at home and in the community. Follow-up surveys performed three months after the performance indicated that participants had discussed the play with others and had increased their participation in substance abuse prevention activities, particularly regarding donations of money. CONCLUSION: Drama incorporates a component of emotional response to the informational content, and the combination of emotion and information works together to promote individual intentions to become more involved in family and community prevention activities. This study demonstrates the efficacy of drama as a mechanism to educate and motivate. Support for this mechanism is warranted at the level of state, local community, school district, and faith-based and community organizations. PMID- 17411424 TI - The emergence of semantic categorization in early visual processing: ERP indices of animal vs. artifact recognition. AB - BACKGROUND: Neuroimaging and neuropsychological literature show functional dissociations in brain activity during processing of stimuli belonging to different semantic categories (e.g., animals, tools, faces, places), but little information is available about the time course of object perceptual categorization. The aim of the study was to provide information about the timing of processing stimuli from different semantic domains, without using verbal or naming paradigms, in order to observe the emergence of non-linguistic conceptual knowledge in the ventral stream visual pathway. Event related potentials (ERPs) were recorded in 18 healthy right-handed individuals as they performed a perceptual categorization task on 672 pairs of images of animals and man-made objects (i.e., artifacts). RESULTS: Behavioral responses to animal stimuli were ~50 ms faster and more accurate than those to artifacts. At early processing stages (120-180 ms) the right occipital-temporal cortex was more activated in response to animals than to artifacts as indexed by posterior N1 response, while frontal/central N1 (130-160) showed the opposite pattern. In the next processing stage (200-260) the response was stronger to artifacts and usable items at anterior temporal sites. The P300 component was smaller, and the central/parietal N400 component was larger to artifacts than to animals. CONCLUSION: The effect of animal and artifact categorization emerged at ~150 ms over the right occipital temporal area as a stronger response of the ventral stream to animate, homomorphic, entities with faces and legs. The larger frontal/central N1 and the subsequent temporal activation for inanimate objects might reflect the prevalence of a functional rather than perceptual representation of manipulable tools compared to animals. Late ERP effects might reflect semantic integration and cognitive updating processes. Overall, the data are compatible with a modality specific semantic memory account, in which sensory and action-related semantic features are represented in modality-specific brain areas. PMID- 17411425 TI - Analysis of NAMCS data for multiple sclerosis, 1998-2004. AB - BACKGROUND: To our knowledge, no study to date has investigated the prescribing patterns of immunomodulatory agents (IMAs) in an outpatient setting in the United States. To address this issue, we performed retrospective data analyses on National Ambulatory Medical Care Survey (NAMCS) data for MS patient visits between 1998 and 2004. METHODS: NAMCS data are a weighted estimate of the nationwide frequency of patients' outpatient clinic visits. We analyzed NAMCS data in the following categories: (1) the proportion of MS patient visits to neurologists, family practitioners or internists, (2) age/gender/race/geographical distribution patterns in patient visits, and (3) the proportion of patients on IMA treatment among established MS patients. RESULTS: There were an estimated 6.7 million multiple sclerosis (MS) patient visits to the clinics between 1998-2004. Neurologists recorded the most patient visits, 50.7%. Patient visits were mostly in the fourth and fifth decade age group (57.9%). The male to female ratio was 1:4. No statistical evidence was observed for a decline or increase in IMA usage. About 62% patients visiting neurologists and 92% seen by family practitioners/internists were not using IMAs. Our results suggest that between the years 1998-2003, the use of interferon-1a tended to decline while the use of interferon-1b and glatiramer acetate, increased. CONCLUSION: Strategies that lead to improved use of IMAs in the management of MS in the outpatient setting are needed. PMID- 17411426 TI - Novel APC mutations in Czech and Slovak FAP families: clinical and genetic aspects. AB - BACKGROUND: Germline mutations in the adenomatous polyposis gene (APC) result in familial adenomatous polyposis (FAP). FAP is an autosomal dominantly inherited disorder predisposing to colorectal cancer. Typical FAP is characterized by hundreds to thousands of colorectal adenomatous polyps and by several extracolonic manifestations. An attenuated form of polyposis (AFAP) is characterized by less than 100 adenomas and later onset of the disease. METHODS: Here, we analyzed the APC gene for germline mutations in 59 Czech and 15 Slovak FAP patients. In addition, 50 apparently APC mutation negative Czech probands and 3 probands of Slovak origin were screened for large deletions encompassing the APC gene. Mutation screening was performed using denaturing gradient gel electrophoresis and/or protein truncation test. DNA fragments showing an aberrant electrophoretic banding pattern were sequenced. Screening for large deletions was performed by multiplex ligation dependent probe amplification. The extent of deletions was analyzed using following microsatellite markers: D5S299, D5S82, D5S134 and D5S346. RESULTS: In the set of Czech and Slovak patients, we identified 46 germline mutations among 74 unrelated probands. Total mutation capture is 62,2% including large deletions. Thirty seven mutations were detected in 49 patients presenting a classical FAP phenotype (75,5%) and 9 mutations in 25 patients with attenuated FAP (36%). We report 20 novel germline APC mutations and 3 large deletions (6%) encompassing the whole-gene deletions and/or exon 14 deletion. In the patients with novel mutations, correlations of the mutation localization are discussed in context of the classical and/or attenuated phenotype of the disease. CONCLUSION: The results of the molecular genetic testing are used both in the establishment of the predictive diagnosis and in the clinical management of patients. In some cases this study has also shown the difficulty to classify clinically between the classical and the attenuated form of FAP according to the established criteria. Interfamilial and/or intrafamilial phenotype variability was also confirmed in some cases which did not fit well with predicted genotype-phenotype correlation. All these findings have to be taken into consideration both in the genetic counselling and in the patient care. PMID- 17411427 TI - Comparative analysis indicates regulatory neofunctionalization of yeast duplicates. AB - BACKGROUND: Gene duplication provides raw material for the generation of new functions, but most duplicates are rapidly lost due to the initial redundancy in gene function. How gene function diversifies following duplication is largely unclear. Previous studies analyzed the diversification of duplicates by characterizing their coding sequence divergence. However, functional divergence can also be attributed to changes in regulatory properties, such as protein localization or expression, which require only minor changes in gene sequence. RESULTS: We developed a novel method to compare expression profiles from different organisms and applied it to analyze the expression divergence of yeast duplicated genes. The expression profiles of Saccharomyces cerevisiae duplicate pairs were compared with those of their pre-duplication orthologs in Candida albicans. Duplicate pairs were classified into two classes, corresponding to symmetric versus asymmetric rates of expression divergence. The latter class includes 43 duplicate pairs in which only one copy has a significant expression similarity to the C. albicans ortholog. These may present cases of regulatory neofunctionalization, as supported also by their dispensability and variability. CONCLUSION: Duplicated genes may diversify through regulatory neofunctionalization. Notably, the asymmetry of gene sequence evolution and the asymmetry of gene expression evolution are only weakly correlated, underscoring the importance of expression analysis to elucidate the evolution of novel functions. PMID- 17411428 TI - Radiation retinopathy: case report and review. AB - BACKGROUND: Ocular damage from radiation treatment is a well established phenomenon. Many factors are now known to influence the incidence of radiation retinopathy, including total dosage and daily fraction size. Patients who are diabetic, hypertensive or received previous chemotherapy are more susceptible to radiation retinopathy. CASE PRESENTATION: A 55 year old male was referred from the oncology department with epiphora. His medical history included Type 2 Insulin treated Diabetes Mellitus and hypertension. One year prior to presentation he had undergone a total rhinectomy with a 4 week course of post operative radiotherapy for an aggressive sqaumous cell carcinoma of the nose. On examination the visual acuity was noted to be 6/36 left eye and 6/9 right eye. Posterior segment examination revealed marked retinal ischaemia present in the posterior pole and macular region of both eyes. The appearance was not thought to be typical of diabetic changes, radiation retinopathy being the more likely diagnosis especially in view of his history. Over the next four months the vision in both eyes rapidly deteriorated to 3/60 left eye and 1/60 right eye. Bilateral pan retinal photocoagulation was thought to be appropriate treatment at this point. CONCLUSION: This case highlights the importance for ophthalmologists and oncologists to be aware of the close relationship between diabetes and radiation treatment and the profound rapid impact this combination of factors may have on visual function. Radiation is being used with increasing frequency for ocular and orbital disease, because of this more cases of radiation retinopathy may become prevalent. Factors which may potentiate radiation retinopathy should be well known including, increased radiation dosage, increased fraction size, concomitant systemic vascular disease and use of chemotherapy. Counselling should be offered in all cases at risk of visual loss. As no effective treatment currently exists to restore visual function, monitoring of visual acuity in all cases and early referral to the ophthalmologist as appropriate is warranted. PMID- 17411429 TI - Higher prevalence of smoking and lower BMI, waist circumference, cholesterol and triacylglyceride levels in Prague's homeless compared to a majority of the Czech population. AB - BACKGROUND: Homeless people have higher morbidity and mortality rates than the general population. Research has shown that cardiovascular disease is the leading cause of death in older homeless adults. This study was undertaken to describe the prevalence of cardiovascular risk factors in the homeless population in Prague. METHODS: Data was obtained from a cross-sectional study carried out in 2003. Body mass index (BMI), waist circumference (WC), total cholesterol (TC), triacylglycerides (TAG) and smoking habits were assessed. The homeless participants in the study were recruited from a homeless center run by a Prague charitable organization called Nadeje ("Hope") and at Prague's main railway station. Most participants were assessed at the Nadeje center (134 persons) while the rest were assessed at Prague's Bulovka University Hospital (67 persons). RESULTS: A total of 201 homeless (174 males and 27 females) aged 19-70 years were examined. Mean values of BMI, WC, TC and TAG in homeless men and women were within normal limits. Compared with the majority of the Czech population, the homeless had significantly lower mean levels of TC and TAG and lower BMI and WC values. When compared to the majority of the Czech population, the incidence of smoking among the homeless was significantly higher. Among smokers in both populations, no differences were found in the number of cigarettes smoked per day. CONCLUSION: Classical cardiovascular risk factors such as TC, TAG, BMI and WC, are significantly lower in Prague's homeless minority than in the majority of the Czech population. However, the prevalence of smoking is much higher in the homeless population. PMID- 17411430 TI - Graduates of Lebanese medical schools in the United States: an observational study of international migration of physicians. AB - BACKGROUND: As healthcare systems around the world are facing increasing physician shortages, more physicians are migrating from low to high income countries. As an illustrative case of international migration of physicians, we evaluated the current number and historical trends of Lebanese medical graduates (LMG) in the US, and compared their characteristics to those of US medical graduates (USMG) and other international medical graduates (IMG). METHODS: We evaluated the number of LMG using the 2004 the American Medical Association Physicians' Professional Data (AMA-PPD) and then compared it to the number of graduates of other countries. We evaluated the historical trends using the 1978 2004 historical files of the AMA-PPD. We analyzed the characteristics of all LMG and compared them to a random sample of 1000 USMG and a random sample of 1000 IMG using the 2004 AMA-PPD. RESULTS: In 2004, there were 2,796 LMG in the US, constituting 1.3% of all IMG. Compared to other foreign countries contributing to the US physician workforce, Lebanon ranked 2nd after adjusting for country population size (about 4 million) and 21st overall. About 40% of those who graduated from Lebanese medical schools in the last 25 years are currently active physicians in the US. Since 1978, the number of LMG in the US showed a consistent upward trend at a rate of approximately 71 additional graduates per year. Compared with USMG and IMG, LMG were more likely to work in medical research (OR = 2.31; 95% Confidence Interval (CI) = 1.21; 4.43 and OR = 2.63; 95% CI = 1.34; 5.01, respectively) and to be board certified (OR = 1.43; 95% CI = 1.14; 1.78 and OR = 2.04; 95% CI = 1.65;2.53, respectively) and less likely to be in family practice (OR = 0.14; 95% CI = 0.10; 0.19 and OR = 0.18; 95% CI = 0.12; 0.26, respectively). CONCLUSION: Given the magnitude and historical trends of migration of LMG to the US, further exploration of its causes and impact is warranted. High income countries should consider the consequences of their human resources policies on both low income countries' and their own healthcare systems. PMID- 17411431 TI - The disarrayed mutation results in cell cycle and neurogenesis defects during retinal development in zebrafish. AB - BACKGROUND: The vertebrate retina is derived from proliferative neuroepithelial cells of the optic cup. During retinal development, cell proliferation and the processes of cell cycle exit and neurogenesis are coordinated in neuroepithelial progenitor cells. Previous studies have demonstrated reciprocal influences between the cell cycle and neurogenesis. However the specific mechanisms and exact relationships of cell cycle regulation and neurogenesis in the vertebrate retina remain largely unknown. RESULTS: We have isolated and characterized a zebrafish mutant, disarrayed (drya64), which exhibits retinal defects in cell cycle regulation and neurogenesis. By 42 hours post fertilization, disarrayed mutants show small eyes and a reduced forebrain. Other aspects of development appear normal. Although retinogenesis is delayed, mutant retinal cells eventually differentiate to all major cell types. Examination of the disarrayed mitotic cycle using BrdU and direct imaging techniques revealed that retinal neuroepithelial cells have an extended cell cycle period and reduced rate of cell cycle exit and neurogenesis, despite the fact that neurogenesis initiates at the appropriate time of development. Genetic mosaic analyses indicate that the cell cycle phenotype of disarrayed is cell-non-autonomous. CONCLUSION: The disarrayed mutant shows defects in both cell cycle regulation and neurogenesis and provides insights into the coordinated regulation of these processes during retinal development. PMID- 17411432 TI - The European charter for counteracting obesity: a late but important step towards action. Observations on the WHO-Europe ministerial conference, Istanbul, November 15-17, 2006. AB - BACKGROUND: On November 15-17, 2006 the World Health Organization Regional Office for Europe organised a ministerial conference on counteracting obesity in the European region. Delegations from 48 countries met in Istanbul, Turkey. Observed by relevant nongovernmental organisations and expert temporary advisors, the European ministers adopted a charter on counteracting obesity. This charter states that countries within the European region should be able to show results in slowing down and stopping the obesity epidemic within the next 4-5 years, especially among children, and that the obesity prevalence trends should be reversed before 2015. To achieve this, the charter explicitly calls for action beyond health education: changes in the physical, political, informational and social environments are needed to facilitate a healthy energy balanced lifestyle. DISCUSSION: The fact that all member states of WHO-Europe have now explicitly agreed on an ecological approach to fighting the obesity epidemic with a timeline for visible results is important. However, the charter does not explicate specific enough and measurable objectives for improvement, nor the means needed to reach these. SUMMARY: The fact that all WHO-Europe member states have agreed on a charter that recognizes that counteracting obesity requires a multidisciplinary and ecological approach, with a timeline for improvements, is a late but important step forward for public health policy and practice across Europe. However, more specific tangible goals should now be set, the required means should be allocated, coordinated and immediate action should be implemented, and research to identify effective strategies should be encouraged and facilitated. PMID- 17411433 TI - Evolution of protein complexes by duplication of homomeric interactions. AB - BACKGROUND: Cellular functions are accomplished by the concerted actions of functional modules. The mechanisms driving the emergence and evolution of these modules are still unclear. Here we investigate the evolutionary origins of protein complexes, modules in physical protein-protein interaction networks. RESULTS: We studied protein complexes in Saccharomyces cerevisiae, complexes of known three-dimensional structure in the Protein Data Bank and clusters of pairwise protein interactions in the networks of several organisms. We found that duplication of homomeric interactions, a large class of protein interactions, frequently results in the formation of complexes of paralogous proteins. This route is a common mechanism for the evolution of complexes and clusters of protein interactions. Our conclusions are further confirmed by theoretical modelling of network evolution. We propose reasons for why this is favourable in terms of structure and function of protein complexes. CONCLUSION: Our study provides the first insight into the evolution of functional modularity in protein protein interaction networks, and the origins of a large class of protein complexes. PMID- 17411434 TI - Annelid phylogeny and the status of Sipuncula and Echiura. AB - BACKGROUND: Annelida comprises an ancient and ecologically important animal phylum with over 16,500 described species and members are the dominant macrofauna of the deep sea. Traditionally, two major groups are distinguished: Clitellata (including earthworms, leeches) and "Polychaeta" (mostly marine worms). Recent analyses of molecular data suggest that Annelida may include other taxa once considered separate phyla (i.e., Echiura, and Sipuncula) and that Clitellata are derived annelids, thus rendering "Polychaeta" paraphyletic; however, this contradicts classification schemes of annelids developed from recent analyses of morphological characters. Given that deep-level evolutionary relationships of Annelida are poorly understood, we have analyzed comprehensive datasets based on nuclear and mitochondrial genes, and have applied rigorous testing of alternative hypotheses so that we can move towards the robust reconstruction of annelid history needed to interpret animal body plan evolution. RESULTS: Sipuncula, Echiura, Siboglinidae, and Clitellata are all nested within polychaete annelids according to phylogenetic analyses of three nuclear genes (18S rRNA, 28S rRNA, EF1alpha; 4552 nucleotide positions analyzed) for 81 taxa, and 11 nuclear and mitochondrial genes for 10 taxa (additional: 12S rRNA, 16S rRNA, ATP8, COX1-3, CYTB, NAD6; 11,454 nucleotide positions analyzed). For the first time, these findings are substantiated using approximately unbiased tests and non-scaled bootstrap probability tests that compare alternative hypotheses. For echiurans, the polychaete group Capitellidae is corroborated as the sister taxon; while the exact placement of Sipuncula within Annelida is still uncertain, our analyses suggest an affiliation with terebellimorphs. Siboglinids are in a clade with other sabellimorphs, and clitellates fall within a polychaete clade with aeolosomatids as their possible sister group. None of our analyses support the major polychaete clades reflected in the current classification scheme of annelids, and hypothesis testing significantly rejects monophyly of Scolecida, Palpata, Canalipalpata, and Aciculata. CONCLUSION: Using multiple genes and explicit hypothesis testing, we show that Echiura, Siboglinidae, and Clitellata are derived annelids with polychaete sister taxa, and that Sipuncula should be included within annelids. The traditional composition of Annelida greatly underestimates the morphological diversity of this group, and inclusion of Sipuncula and Echiura implies that patterns of segmentation within annelids have been evolutionarily labile. Relationships within Annelida based on our analyses of multiple genes challenge the current classification scheme, and some alternative hypotheses are provided. PMID- 17411435 TI - Beet necrotic yellow vein virus accumulates inside resting spores and zoosporangia of its vector Polymyxa betae BNYVV infects P. betae. AB - BACKGROUND: Plasmodiophorids and chytrids are zoosporic parasites of algae and land plant and are distributed worldwide. There are 35 species belonging to the order Plasmodiophorales and three species, Polymyxa betae, P. graminis, and Spongospora subterranea, are plant viral vectors. Plasmodiophorid transmitted viruses are positive strand RNA viruses belonging to five genera. Beet necrotic yellow vein virus (BNYVV) and its vector, P. betae, are the causal agents for rhizomania. RESULTS: Evidence of BNYVV replication and movement proteins associating with P. betae resting spores was initially obtained using immunofluorescence labeling and well characterized antisera to each of the BNYVV proteins. Root cross sections were further examined using immunogold labeling and electron microscopy. BNYVV proteins translated from each of the four genomic and subgenomic RNAs accumulate inside P. betae resting spores and zoospores. Statistical analysis was used to determine if immunolabelling detected viral proteins in specific subcellular domains and at a level greater than in control samples. CONCLUSION: Virus-like particles were detected in zoosporangia. Association of BNYVV replication and movement proteins with sporangial and sporogenic stages of P. betae suggest that BNYVV resides inside its vector during more than one life cycle stage. These data suggest that P. betae might be a host as well as a vector for BNYVV. PMID- 17411436 TI - The effect of different public health interventions on longevity, morbidity, and years of healthy life. AB - BACKGROUND: Choosing cost-effective strategies for improving the health of the public is difficult because the relative effects of different types of interventions are not well understood. The benefits of one-shot interventions may be different from the benefits of interventions that permanently change the probability of getting sick, recovering, or dying. Here, we compare the benefits of such types of public health interventions. METHODS: We used multi-state life table methods to estimate the impact of five types of interventions on mortality, morbidity (years of life in fair or poor health), and years of healthy life (years in excellent, very good, or good health). RESULTS: A one-shot intervention that makes all the sick persons healthy at baseline would increase life expectancy by 3 months and increase years of healthy life by 6 months, in a cohort beginning at age 65. An equivalent amount of improvement can be obtained from an intervention that either decreases the probability of getting sick each year by 12%, increases the probability of a sick person recovering by 16%, decreases the probability that a sick person dies by 15%, or decreases the probability that a healthy person dies by 14%. Interventions aimed at keeping persons healthy increased longevity and years of healthy life, while decreasing morbidity and medical expenditures. Interventions focused on preventing mortality had a greater effect on longevity, but had higher future morbidity and medical expenditures. Results differed for older and younger cohorts and depended on the value to society of an additional year of sick life. CONCLUSION: Interventions that promote health and prevent disease performed well, but other types of intervention were sometimes better. The value to society of interventions that increase longevity but also increase morbidity needs further research. More comprehensive screening and treatment of new Medicare enrollees might improve their health and longevity without increasing future medical expenditures. PMID- 17411437 TI - Pattern of neuropsychological performance among HIV positive patients in Uganda. AB - BACKGROUND: Few studies have examined cognitive functioning of HIV positive patients in sub-Saharan Africa. It cannot be assumed that HIV positive patients in Africa exhibit the same declines as patients in high-resource settings, since there are differences that may influence cognitive functioning including nutrition, history of concomitant disease, and varying HIV strains, among other possibilities. Part of the difficulty of specifying abnormalities in neuropsychological functioning among African HIV positive patients is that there are no readily available African normative databases. The purpose of the current study was to evaluate the pattern of neuropsychological performance in a sample of HIV positive patients in comparison to HIV negative control subjects in Uganda. METHODS: The neuropsychological test scores of 110 HIV positive patients (WHO Stage 2, n = 21; WHO Stage 3, n = 69; WHO Stage 4, n = 20) were contrasted with those of 100 control subjects on measures of attention/concentration, mental flexibility, learning/memory, and motor functioning. RESULTS: Analysis of covariance (ANCOVA) revealed significant group differences on measures of verbal learning and memory, speed of processing, attention and executive functioning between HIV seropositive and seronegative subjects. CONCLUSION: Ugandan patients with HIV demonstrated relative deficits on measures of verbal learning and memory, speed of processing, attention, and executive functioning compared to HIV negative controls. These results from a resource limited region where clades A and D are prevalent are consistent with previous findings in the developed world where clade B predominates. PMID- 17411438 TI - Impact of AtNHX1, a vacuolar Na+/H+ antiporter, upon gene expression during short and long-term salt stress in Arabidopsis thaliana. AB - BACKGROUND: AtNHX1, the most abundant vacuolar Na+/H+ antiporter in Arabidopsis thaliana, mediates the transport of Na+ and K+ into the vacuole, influencing plant development and contributing to salt tolerance. In this report, microarray expression profiles of wild type plants, a T-DNA insertion knockout mutant of AtNHX1 (nhx1), and a 'rescued' line (NHX1::nhx1) were exposed to both short (12 h and 48 h) and long (one and two weeks) durations of a non-lethal salt stress to identify key gene transcripts associated with the salt response that are influenced by AtNHX1. RESULTS: 147 transcripts showed both salt responsiveness and a significant influence of AtNHX1. Fifty-seven of these genes showed an influence of the antiporter across all salt treatments, while the remaining genes were influenced as a result of a particular duration of salt stress. Most (69%) of the genes were up-regulated in the absence of AtNHX1, with the exception of transcripts encoding proteins involved with metabolic and energy processes that were mostly down-regulated. CONCLUSION: While part of the AtNHX1-influenced transcripts were unclassified, other transcripts with known or putative roles showed the importance of AtNHX1 to key cellular processes that were not necessarily limited to the salt stress response; namely calcium signaling, sulfur metabolism, cell structure and cell growth, as well as vesicular trafficking and protein processing. Only a small number of other salt-responsive membrane transporter transcripts appeared significantly influenced by AtNHX1. PMID- 17411439 TI - High level expression of soluble glycoproteins in the allantoic fluid of embryonated chicken eggs using a Sendai virus minigenome system. AB - BACKGROUND: Embryonated chicken eggs have been used since the mid-20th century to grow a wide range of animal viruses to high titers. However, eggs have found so far only limited use in the production of recombinant proteins. We now describe a system, based on a Sendai virus minigenome, to produce large amounts of heterologous viral glycoproteins in the allantoic cavity of embryonated eggs. RESULTS: Soluble forms of human respiratory syncytial virus (HRSV) and human metapneumovirus (HMPV) fusion (F) proteins, devoid of their transmembrane and cytoplasmic domains, were produced in allantoic fluids using the Sendai minigenome system. The first step was rescuing in cell cultures Sendai virus minigenomes encoding the proteins of interest, with the help of wild type Sendai virus. The second step was propagating such recombinant defective viruses, together with the helper virus, in the allantoic cavity of chicken embryonated eggs, and passage to optimize protein production. When compared with the production of the same proteins in the culture supernatant of cells infected with vaccinia recombinants, the yield in the allantoic fluid was 5-10 fold higher. Mutant forms of these soluble proteins were easily constructed by site-directed mutagenesis and expressed in eggs using the same approach. CONCLUSION: The simplicity and economy of the Sendai minigenome system, together with the high yield achieved in the allantoic fluid of eggs, makes it an attractive method to express soluble glycoproteins aimed for structural studies. PMID- 17411440 TI - Ovarian cancer risk and common variation in the sex hormone-binding globulin gene: a population-based case-control study. AB - BACKGROUND: The sex hormone-binding globulin (SHBG) is a carrier protein that modulates the bio-availability of serum sex steroid hormones, which may be involved in ovarian cancer. We evaluated whether common genetic variation in SHBG and its 3' neighbor ATP1B2, in linkage disequilibrium, is associated with the risk of epithelial ovarian cancer. METHODS: The study population included 264 women with ovarian carcinoma and 625 controls participating in a population-based case-control study in Poland. Five common single nucleotide polymorphisms (SNPs) in SHGB and five in ATP1B2 were selected to capture most common variation in this region. RESULTS: None of the SNPs evaluated was significantly associated with ovarian cancer risk, including the putative functional SNPs SHBG D356N (rs6259) and -67G>A 5'UTR (rs1799941). However, our data were consistent with a decreased ovarian cancer risk associated with the variant alleles for these two SNPs, which have been previously associated with increased circulating levels of SHBG. CONCLUSION: These data do not support a substantial association between common genetic variation in SHBG and ovarian cancer risk. PMID- 17411441 TI - Perlecan controls neurogenesis in the developing telencephalon. AB - BACKGROUND: Perlecan is a proteoglycan expressed in the basal lamina of the neuroepithelium during development. Perlecan absence does not impair basal lamina assembly, although in the 55% of the mutants early disruptions of this lamina conducts to exencephaly, impairing brain development. The rest of perlecan-null brains complete its prenatal development, maintain basal lamina continuity interrupted by some isolated ectopias, and are microcephalic. Microcephaly consists of thinner cerebral walls and underdeveloped ganglionic eminences. We have studied the mechanisms that generate brain atrophy in telencephalic areas where basal lamina is intact. RESULTS: Brain atrophy in the absence of perlecan started in the ventral forebrain and extended to lateral and dorsal parts of the cortex in the following stages. First, the subpallial forebrain developed poorly in early perlecan-null embryos, because of a reduced cell proliferation: the number of cells in mitosis decreased since the early stages of development. This reduction resulted in a decreased tangential migration of interneurons to the cerebral cortex. Concomitant with the early hypoplasia observed in the medial ganglionic eminences, Sonic Hedgehog signal decreased in the perlecan-null floor plate basal lamina at E12.5. Second, neurogenesis in the pallial neuroepithelium was affected in perlecan deficient embryos. We found reductions of nearly 50% in the number of cells exiting the cell cycle at E12-E13. The labeling index, which was normal at this age, significantly decreased with advancing corticogenesis. Moreover, nestin+ or PCNA+ progenitors increased since E14.5, reaching up to about 150% of the proportion of PCNA+ cells in the wild-type at E17.5. Thus, labeling index reduction together with increased progenitor population, suggests that atrophy is the result of altered cell cycle progression in the cortical progenitors. Accordingly, less neurons populated the cortical plate and subplate of perlecan-null neocortex, as seen with the neuronal markers beta-tubulin and Tbr1. CONCLUSION: As a component of the basal lamina, perlecan both maintains this structure and controls the response of the neuroepithelium to growth factors. Less mitotic cells in the early medial ganglionic eminences, and impaired cell cycle progression in the late neocortex, suggests insufficient recruitment and signaling by neurogenic morphogens, such as SHH or FGF2. PMID- 17411442 TI - Culture-independent analysis of bacterial diversity in a child-care facility. AB - BACKGROUND: Child-care facilities appear to provide daily opportunities for exposure and transmission of bacteria and viruses. However, almost nothing is known about the diversity of microbial contamination in daycare facilities or its public health implications. Recent culture-independent molecular studies of bacterial diversity in indoor environments have revealed an astonishing diversity of microorganisms, including opportunistic pathogens and many uncultured bacteria. In this study, we used culture and culture-independent methods to determine the viability and diversity of bacteria in a child-care center over a six-month period. RESULTS: We sampled surface contamination on toys and furniture using sterile cotton swabs in four daycare classrooms. Bacteria were isolated on nutrient and blood agar plates, and 16S rRNA gene sequences were obtained from unique (one of a kind) colony morphologies for species identification. We also extracted DNA directly from nine representative swab samples taken over the course of the study from both toy and furniture surfaces, and used "universal" 16S rRNA gene bacterial primers to create PCR-based clone libraries. The rRNA gene clones were sequenced, and the sequences were compared with related sequences in GenBank and subjected to phylogenetic analyses to determine their evolutionary relationships. Culturing methods identified viable bacteria on all toys and furniture surfaces sampled in the study. Bacillus spp. were the most commonly cultured bacteria, followed by Staphylococcus spp., and Microbacterium spp. Culture-independent methods based on 16S rRNA gene sequencing, on the other hand, revealed an entirely new dimension of microbial diversity, including an estimated 190 bacterial species from 15 bacterial divisions. Sequence comparisons and phylogenetic analyses determined that the clone libraries were dominated by a diverse set of sequences related to Pseudomonas spp., as well as uncultured bacteria originally identified on human vaginal epithelium. Other sequences were related to uncultured bacteria from wastewater sludge, and many human-associated bacteria including a number of pathogens and opportunistic pathogens. Our results suggest that the child-care facility provided an excellent habitat for slime producing Pseudomonads, and that diaper changing contributed significantly to the bacterial contamination. CONCLUSION: The combination of culture and culture independent methods provided powerful means for determining both viability and diversity of bacteria in child-care facilities. Our results provided insight into the source of contamination and suggested ways in which sanitation might be improved. Although our study identified a remarkable array of microbial diversity present in a single daycare, it also revealed just how little we comprehend the true extent of microbial diversity in daycare centers or other indoor environments. PMID- 17411443 TI - Meta-analysis of several gene lists for distinct types of cancer: a simple way to reveal common prognostic markers. AB - BACKGROUND: Although prognostic biomarkers specific for particular cancers have been discovered, microarray analysis of gene expression profiles, supported by integrative analysis algorithms, helps to identify common factors in molecular oncology. Similarities of Ordered Gene Lists (SOGL) is a recently proposed approach to meta-analysis suitable for identifying features shared by two data sets. Here we extend the idea of SOGL to the detection of significant prognostic marker genes from microarrays of multiple data sets. Three data sets for leukemia and the other six for different solid tumors are used to demonstrate our method, using established statistical techniques. RESULTS: We describe a set of significantly similar ordered gene lists, representing outcome comparisons for distinct types of cancer. This kind of similarity could improve the diagnostic accuracies of individual studies when SOGL is incorporated into the support vector machine algorithm. In particular, we investigate the similarities among three ordered gene lists pertaining to mesothelioma survival, prostate recurrence and glioma survival. The similarity-driving genes are related to the outcomes of patients with lung cancer with a hazard ratio of 4.47 (p = 0.035). Many of these genes are involved in breakdown of EMC proteins regulating angiogenesis, and may be used for further research on prognostic markers and molecular targets of gene therapy for cancers. CONCLUSION: The proposed method and its application show the potential of such meta-analyses in clinical studies of gene expression profiles. PMID- 17411444 TI - Reactive arthritis developing after pneumococcal conjunctivitis: a case report. AB - BACKGROUND: S. pneumoniae is a known cause of bacterial conjunctivitis and can be transmitted through contact with infected carriers. CASE PRESENTATION: A 38-year old ophthalmologist developing reactive arthritis following clinic-acquired pneumococcal conjunctivitis. CONCLUSION: (1) Despite the frequency and largely self-limiting nature of infective conjunctivitis, it should be appropriately assessed and managed, as the natural history can occasionally be associated with significant morbidity. (2) Hygienic measures are required to be implemented by both patients and ophthalmic staff to reduce the likelihood of transmission. PMID- 17411445 TI - Iatrogenic insertion of impression mould into middle ear and mastoid and its retrieval after 9 years: a case report. AB - The magnitude of hearing loss in Pakistan is enormous. One in twelve children of Pakistan suffers from some form of hearing impairment. Many of them are unable to afford surgical procedures and resort to the use of cheap hearing aids fitted by untrained individuals or people lacking the required expertise. This predisposes the patients to significant complications during a process that is otherwise considered safe.We report the case of a child, where the process of making the mould for a hearing aid led to the perforation of the tympanic membrane and pouring of mould material into the middle ear, necessitating surgical intervention. During initial surgery it was thought that all mould had been removed from the middle ear but 9 years later this child underwent cochlear implantation at the same center and remaining part of ear mould was discovered from mastoid cavity. PMID- 17411446 TI - Introducing journal of medical case reports. PMID- 17411447 TI - Acute disseminated encephalomyelitis mimicking late CNS relapse of acute lymphoblastic leukaemia: case report. AB - BACKGROUND: Acute encephalomyelopathy occurring after an allogeneic bone marrow transplant for leukaemia is a diagnostic emergency. The diagnosis can be challenging since there is a wide set of alternative diagnoses, including opportunistic infections and relapse of the leukaemia. CASE PRESENTATION: A 13 year old girl presented with a severe acute myelopathy and encephalopathy. She was in prolonged remission from a central nervous system and bone marrow relapse of an acute lymphoblastic leukaemia, treated with allogeneic bone marrow transplantation. Neuroimaging showed multifocal grey and white matter lesions of demyelinating appearance in the brain and entire spine. Immunophenotyping and cytogenetic investigations of the girl's cerebrospinal fluid lymphocytosis excluded a late central nervous system relapse of her leukaemia. The diagnosis was acute disseminated encephalomyelitis. With standard immunosuppressive therapy, the girl had early cerebral recovery but a prolonged period of recovery from her myelopathy. CONCLUSION: Acute disseminated encephalomyelitis should be considered in the differential diagnosis of acute encephalomyelopathy after bone marrow transplantation for leukaemia. Demyelinating syndromes such as acute disseminated encephalomyelitis may be late sequelae of bone marrow transplantation. PMID- 17411448 TI - Dissection and hemostasis with hydroxilated polyvinyl acetal tampons in open thyroid surgery. AB - BACKGROUND: The essential objectives for thyroidectomy are: avoidance of injury to the recurrent laryngeal nerves, conservation of the parathyroid glands, an accurate haemostasis and an excellent cosmesis. In the last 10 years major improvements and new technologies have been proposed and applied in thyroid surgery; among these mini-invasive thyroidectomy, regional anaesthesia and intraoperative neuromonitoring, and new devices for achieving dissection and haemostasis. Minor bleeding from small vessels could be a major complication in thyroid surgery. The purpose of ligating vessels is to maintain the surgical site free from an excess of blood and reduce blood loss in the patient. MATERIALS AND METHODS: Hydroxylated polyvinyl acetal tampons (HPA) are made by a synthetic, open cell foam structure able to absorb fluids up to 25 times the initial weight. We tested their efficacy for small bleeding control and tissue dissection during several thyroid procedures. RESULTS: HPA tampons have been found extremely useful to absorb blood coming from minor and diffuse loss, helping to control bleeding by a combined action of fluid absorption and local compression. The porous design of the tampon allows the use of the suction device right through the tampon itself. Thanks to the initial mildly hard consistency, we also used HPA tampons as dissecting instruments. CONCLUSION: In our experience the use of HPA tampons resulted extremely efficient for minor bleeding control, fluids removal and tissue dissection during thyroid surgery. PMID- 17411449 TI - Non-invasive MR imaging of inflammation in a patient with both asymptomatic carotid atheroma and an abdominal aortic aneurysm: a case report. AB - Inflammation is a recognized risk factor for the vulnerable atherosclerotic plaque. USPIO-enhanced MRI imaging is a promising non-invasive method to identify high-risk atheromatous plaque inflammation in vivo in humans, in which areas of focal signal loss on MR images have been shown to correspond to the location of activated macrophages, typically at the shoulder regions of the plaque. This is the first report in humans describing simultaneous USPIO uptake within atheroma in two different arterial territories and again emphasises that atherosclerosis is a truly systemic disease. With further work, USPIO-enhanced MR imaging may be useful in identifying inflamed vulnerable atheromatous plaques in vivo, so refining patient selection for intervention and allowing appropriate early aggressive pharmacotherapy to prevent plaque rupture. PMID- 17411450 TI - The influence of psychological factors on the outcomes of laparoscopic Nissen fundoplication. AB - BACKGROUND: Psychological factors play a role in a variety of gastrointestinal illness, including gastroesophageal reflux disease (GERD). Their impact on the surgical outcomes of antireflux surgery is unknown. METHODS: This is a single institution prospective controlled trial, comparing patients undergoing a laparoscopic Nissen fundoplication for GERD (LNF Group, n = 17) to patients undergoing an elective laparoscopic cholecystectomy for biliary colic (Control Group, n = 10). All patients had a psychological assessment before surgery, at 3 months and 6 months after surgery (i.e. Symptom CheckList-90-R somatization subset (SCL-90-R), Depression Anxiety Stress Scales, Anxiety sensitivity index, Illness attitude scale and Beck Depression Inventory II). GERD symptoms were recorded in the LNF Group using a standardized questionnaire (score 0-60). Patients with post-operative GERD symptoms score above 12 at 6 months were evaluated specifically. Statistical analysis was performed using a Student T test, and statistical significance was set at 0.05. RESULTS: There was no significant difference in preoperative and postoperative psychological assessment between the two groups. In the LNF Group, 7 patients had persisting GERD symptoms at 6 months (GERD score greater than 12). The preoperative SCL-90-R score was also significantly higher in this subgroup, when compared to the rest of the LNF Group (18.2 versus 8.3, p < 0.05) and to the Control Group (18.2 versus 7.9, p < 0.05). There was no significant difference for the other psychological tests. CONCLUSION: The SCL-90-R Somatization Subset, reflecting the level of somatization in a patient, may be useful to predict poor outcomes after antireflux surgery. Cognizance of psychological disorders could improve the selection of an optimal treatment for GERD and help reduce the rate of ongoing symptoms after LNF. PMID- 17411451 TI - Toxic shock syndrome responsive to steroids. AB - BACKGROUND: Toxic Shock Syndrome is a dangerous disease with clinical features mimicking bacterial sepsis. The best management of Toxic Shock Syndrome is not determined. CASE PRESENTATION: A 28 year-old woman presenting with high fever, tachycardia and widespread erythroderma is described. She failed to respond to intravenous antibiotics and required ITU admission. High dose corticosteroids dramatically improved her clinical condition. CONCLUSION: Toxic Shock Syndrome should be considered in the differential diagnosis of unexplained fever, rash and features resembling septic shock. Corticosteroids should be considered in the treatment of Toxic Shock Syndrome. PMID- 17411453 TI - Conflict and health: a paradigm shift in global health and human rights. PMID- 17411454 TI - Acute compartment syndrome of the hand in Henoch-Schonlein Purpura. AB - An eight year old boy with Henoch-Schonlein Purpura (HSP) presented with acute compartment syndrome (ACS) of his left hand following arterial cannulation of his radial artery in intensive care unit. Emergency decompression and fasciotomy were performed. The authors report this first case in literature and discuss how HSP can be complicated by ACS and ways to prevent the latter from happening. PMID- 17411455 TI - HIV-1 prevalence and factors associated with infection in the conflict-affected region of North Uganda. AB - BACKGROUND: Since 1986, northern Uganda has been severely affected by civil strife with most of its population currently living internally displaced in protected camps. This study aims at estimating the HIV-1 prevalence among this population and the factors associated with infection. METHODS: In June-December 2005, a total of 3051 antenatal clinics attendees in Gulu, Kitgum and Pader districts were anonymously tested for HIV-1 infection as part of routine sentinel surveillance. Factors associated with the infection were evaluated using logistic regression models. RESULTS: The age-standardised HIV-1 prevalence was 10.3%, 9.1% and 4.3% in the Gulu, Kitgum and Pader district, respectively. The overall prevalence in the area comprised of these districts was 8.2% when data was weighted according to the districts' population size. Data from all sites combined show that, besides older women [20-24 years: adjusted odds ratio (AOR) = 1.96, 95% confidence interval (CI): 1.29-2.97; 25-29 years: AOR = 2.01, 95% CI: 1.30-3.11; > or = 30 years: AOR = 1.91, 95% CI: 1.23-2.97], unmarried women (AOR = 1.47, 95% CI: 1.06-2.04), and those with a partner with a non-traditional occupation (AOR = 1.62, 95% CI: 1.18-2.21), women living outside of protected camps for internally displaced persons have a higher risk of being HIV-1 infected than internally displaced women (AOR = 1.55, 95% CI: 1.15-2.08). CONCLUSION: Although published data from Gulu district show a declining HIV-1 prevalence trend that is consistent with that observed at the national level since 1993, the prevalence in North Uganda is still high. Internally displaced women have a lower risk of being infected probably because of their reduced mobility and accessibility, and increased access to health prevention services. PMID- 17411456 TI - The trauma of ongoing conflict and displacement in Chechnya: quantitative assessment of living conditions, and psychosocial and general health status among war displaced in Chechnya and Ingushetia. AB - BACKGROUND: Conflict in Chechnya has resulted in over a decade of violence, human rights abuses, criminality and poverty, and a steady flow of displaced seeking refuge throughout the region. At the beginning of 2004 MSF undertook quantitative surveys among the displaced populations in Chechnya and neighbouring Ingushetia. METHODS: Surveys were carried out in Ingushetia (January 2004) and Chechnya (February 2004) through systematic sampling. Various conflict-related factors contributing to ill health were researched to obtain information on displacement history, living conditions, and psychosocial and general health status. RESULTS: The average length of displacement was five years. Conditions in both locations were poor, and people in both locations indicated food shortages (Chechnya (C): 13.3%, Ingushetia (I): 11.3%), and there was a high degree of dependency on outside help (C: 95.4%, I: 94.3%). Most people (C: 94%, I: 98%) were confronted with violence in the past. Many respondents had witnessed the killing of people (C: 22.7%, I: 24.1%) and nearly half of people interviewed witnessed arrests (C: 53.1%, I: 48.4%) and maltreatment (C: 56.2%, I: 44.5%). Approximately one third of those interviewed had directly experienced war-related violence. A substantial number of people interviewed - one third in Ingushetia (37.5%) and two-thirds in Chechnya (66.8%) - rarely felt safe. The violence was ongoing, with respondents reporting violence in the month before the survey (C: 12.5%, I: 4.6%). Results of the general health questionnaire (GHQ 28) showed that nearly all internally displaced persons interviewed were suffering from health complaints such as somatic complaints, anxiety/insomnia, depressive feelings or social dysfunction (C: 201, 78.5%, CI: 73.0% - 83.4%; I: 230, 81.3%, CI: 76.2% - 85.6%). Poor health status was reflected in other survey questions, but health services were difficult to access for around half the population (C: 54.3%, I: 46.6%). DISCUSSION: The study demonstrates that the health needs of internally displaced in both locations are similarly high and equally unaddressed. The high levels of past confrontation with violence and ongoing exposure in both locations is likely to contribute to a further deterioration of the health status of internally displaced. As of March 2007, concerns remain about how the return process is being managed by the authorities. PMID- 17411457 TI - Association between expatriation and HIV awareness and knowledge among injecting drug users in Kabul, Afghanistan: A cross-sectional comparison of former refugees to those remaining during conflict. AB - BACKGROUND: Little is known about human immunodeficiency virus (HIV) awareness among Afghan injecting drug users (IDUs), many of whom initiated injecting as refugees. We explored whether differences in HIV awareness and knowledge exist between Afghan IDUs who were refugees compared to those never having left Afghanistan. METHODS: A convenience sample of IDUs in Kabul, Afghanistan was recruited into a cross-sectional study through street outreach over a one year period beginning in 2005. Participants completed an interviewer-administered questionnaire and underwent voluntary counseling and testing for HIV, syphilis, hepatitis B surface antigen, and hepatitis C antibody. Differences in HIV awareness and specific HIV knowledge between IDU who lived outside the country in the last decade versus those who had not were assessed with logistic regression. RESULTS: Of 464 IDUs, 463 (99%) were male; median age and age at first injection were 29 and 25 years, respectively. Most (86.4%) had lived or worked outside the country in the past ten years. Awareness of HIV was reported by 46.1%; those having been outside the country in the last decade were significantly more likely to have heard of HIV (48.3% vs. 31.7%; OR = 2.00, 95% CI: 1.14 - 3.53). However, of those aware of HIV, only 38.3% could name three correct transmission routes; specific HIV knowledge was not significantly associated with residence outside the country. CONCLUSION: Accurate HIV knowledge among Afghan IDUs is low, though former refugees had greater HIV awareness. Reported high-risk injecting behavior was not significantly different between IDU that were refugees and those that did not leave the country, indicating that all Afghan IDU should receive targeted prevention programming. PMID- 17411458 TI - The boy who refused an IV: a case report of subcutaneous clodronate for bone pain in a child with Ewing Sarcoma. AB - BACKGROUND: Bone pain in malignancy can be challenging to treat. Bisphosphonates have been found to be useful in adults with bone pain, but there are no reports of their use in children for this indication. In pediatric palliative medicine there are hurdles in translating knowledge gained primarily in adult studies into application in children. Obstacles exist in initially determining whether the evidence supports using a drug in children, and once a drug is chosen, then determining the optimal route of delivery. There is very little data to guide pediatric practitioners in this situation. CASE PRESENTATION: A 9 year old boy with disseminated Ewing Sarcoma presented with extremity pain not responsive to a combination of opiates, gabapentin and non-steroidal anti-inflammatory drugs. Clodronate, a bisphosphonate, was added to the regimen to treat bone pain. It was given subcutaneously every 4 weeks with a good response and no side effects. CONCLUSION: This case report describes the use of a bisphosphonate, clodronate, given subcutaneously to a child with Ewing sarcoma with effective relief of bone pain. It describes how the care team encountered the challenges inherent in translating adult therapy into a pediatric regimen. Furthermore the report details how a regimen was developed to address this child's concerns regarding medication administration. Further effort needs to be made at finding solutions to address the lack of good evidence for pediatric palliative therapies. PMID- 17411459 TI - Acute small bowel obstruction as a result of a Meckel's diverticulum encircling the terminal ileum: A case report. AB - BACKGROUND: In the developed world, small bowel obstruction accounts for 20% of all acute surgical admissions. The aetiology for majority of these cases includes postoperative adhesions and herniae. However, a relatively uncommon cause is a Meckel's diverticulum. Although this diagnosis is primarily reported in the adolescent population, it should also be considered in adults. CASE PRESENTATION: In the present report, we present a rare case where a fit and healthy 74-year-old gentleman, with no previous history of abdominal surgery, presented with the cardinal symptoms and signs of small bowel obstruction as the result of a Meckel's diverticulum encircling his terminal ileum. Initial investigations included a supine abdominal x-ray showing dilated loops of small bowel and computerised tomographic imaging of the abdomen, which revealed a stricture in the terminal ileum of unknown aetiology. At laparotomy, multiple loops of distended small bowel were seen from the duodeno-jeujenal junction to the terminal ileum, which was encircled by a Meckel's diverticulum. The Meckel's diverticulum was then divided to release the obstruction, mobilised and subsequently removed. Finally, the small bowel contents were decompressed into the stomach and the nasogastric tube aspirated, before returning the loops of bowel into the abdomen in sequence. The patient made a good postoperative recovery and was discharged home 5 days later. CONCLUSION: This report highlights the importance of considering a Meckel's diverticulum as a cause of small bowel obstruction in individuals from all age groups and especially in a person with no previous abdominal pathology or surgery. PMID- 17411460 TI - Occurrence and overlap of natural disasters, complex emergencies and epidemics during the past decade (1995-2004). AB - BACKGROUND: The fields of expertise of natural disasters and complex emergencies (CEs) are quite distinct, with different tools for mitigation and response as well as different types of competent organizations and qualified professionals who respond. However, natural disasters and CEs can occur concurrently in the same geographic location, and epidemics can occur during or following either event. The occurrence and overlap of these three types of events have not been well studied. METHODS: All natural disasters, CEs and epidemics occurring within the past decade (1995-2004) that met the inclusion criteria were included. The largest 30 events in each category were based on the total number of deaths recorded. The main databases used were the Emergency Events Database for natural disasters, the Uppsala Conflict Database Program for CEs and the World Health Organization outbreaks archive for epidemics. ANALYSIS: During the past decade, 63% of the largest CEs had > or =1 epidemic compared with 23% of the largest natural disasters. Twenty-seven percent of the largest natural disasters occurred in areas with > or = 1 ongoing CE while 87% of the largest CEs had > or = 1 natural disaster. CONCLUSION: Epidemics commonly occur during CEs. The data presented in this article do not support the often-repeated assertion that epidemics, especially large-scale epidemics, commonly occur following large-scale natural disasters. This observation has important policy and programmatic implications when preparing and responding to epidemics. There is an important and previously unrecognized overlap between natural disasters and CEs. Training and tools are needed to help bridge the gap between the different type of organizations and professionals who respond to natural disasters and CEs to ensure an integrated and coordinated response. PMID- 17411461 TI - A possible new syndrome with growth-hormone secreting pituitary adenoma, colonic polyposis, lipomatosis, lentigines and renal carcinoma in association with familial testicular germ cell malignancy: A case report. AB - BACKGROUND: Germ-cell testicular cancer has not been definitively linked to any known hereditary cancer susceptibility disorder. Familial testicular cancer in the presence of other findings in affected and unaffected family members might indicate a previously-unidentified hereditary cancer syndrome. CASE PRESENTATION: The patient was diagnosed with a left testicular seminoma at age 28, and treated with left orchiectomy followed by adjuvant cobalt radiation. His family history is significant for testicular seminoma in his son, bladder cancer in his sister, and lipomatosis in his father. His evaluation as part of an etiologic study of familial testicular cancer revealed multiple colon polyps (adenomatous, hyperplastic, and hamartomatous) first found in his 50 s, multiple lipomas, multiple hyperpigmented skin lesions, left kidney cancer diagnosed at age 64, and a growth-hormone producing pituitary adenoma with associated acromegaly diagnosed at age 64. The patient underwent genetic testing for Cowden syndrome (PTEN gene), Carney complex (PRKAR1A gene), and multiple endocrine neoplasia syndrome type 1 (MEN1 gene); no deleterious mutations were identified. DISCUSSION: The constellation of benign and malignant neoplasms in the context of this patient's familial testicular cancer raised the possibility that these might be manifestations of a known hereditary susceptibility cancer syndrome; however, genetic testing for the three syndromes that were most likely to explain these findings did not show any mutation. Alternatively, this family's phenotype might represent a novel neoplasm susceptibility disorder. This possibility cannot be evaluated definitively on the basis of a single case report; additional observations and studies are necessary to investigate this hypothesis further. PMID- 17411462 TI - How does the health and well-being of young Australian vegetarian and semi vegetarian women compare with non-vegetarians? AB - OBJECTIVE: To compare the sociodemographic characteristics, health status and health service use of vegetarians, semi-vegetarians and non-vegetarians. DESIGN: In cross-sectional data analyses of the Australian Longitudinal Study on Women's Health in 2000, 9113 women (aged 22-27 years) were defined as non-vegetarians if they reported including red meat in their diet, as semi-vegetarians if they excluded red meat and as vegetarians if they excluded meat, poultry and fish from their diet. RESULTS: The estimated prevalence was 3% and 10% for vegetarian and semi-vegetarian young women. Compared with non-vegetarians, vegetarians and semi vegetarians were more likely to live in urban areas and to not be married. Vegetarians and semi-vegetarians had lower body mass index (mean (95% confidence interval): 22.2 (21.7-22.7) and 23.0 (22.7-23.3) kg m(-2)) than non-vegetarians (23.7 (23.6-23.8) kg m(-2)) and tended to exercise more. Semi-vegetarians and vegetarians had poorer mental health, with 21-22% reporting depression compared with 15% of non-vegetarians (P < 0.001). Low iron levels and menstrual symptoms were also more common in both vegetarian groups. Vegetarian and semi-vegetarian women were more likely to consult alternative health practitioners and semi vegetarians reported taking more prescription and non-prescription medications. Compared with non-vegetarians, semi-vegetarians were less likely and vegetarians much less likely to be taking the oral contraceptive pill. CONCLUSION: The levels of physical activity and body mass indices of the vegetarian and semi-vegetarian women suggest they are healthier than non-vegetarians. However, the greater reports of menstrual problems and the poorer mental health of these young women may be of clinical significance. PMID- 17411463 TI - The effects of a middle-school healthy eating intervention on adolescents' fat and fruit intake and soft drinks consumption. AB - OBJECTIVES: To evaluate the effects of a middle-school healthy eating promotion intervention combining environmental changes and computer-tailored feedback, with and without an explicit parent involvement component. DESIGN: Clustered randomised controlled trial. SETTING: Fifteen West-Flemish (Belgian) middle schools. SUBJECTS: A random sample of 15 schools with 2991 pupils in 7th and 8th grades was randomly assigned to an intervention group with parental support (n = 5), an intervention group without parental support (n = 5) and a control group (n = 5). In these 15 schools an intervention combining environmental changes with computer-tailored feedback was implemented. Fat and fruit intake, water and soft drinks consumption were measured with food-frequency questionnaires in the total sample of children. RESULTS: In girls, fat intake and percentage of energy from fat decreased significantly more in the intervention group with parental support, compared with the intervention alone group (all F>3.9, P < 0.05) and the control group (all F>16.7, P < 0.001). In boys, there were no significant decreases in fat intake (F = 1.4, not significant (NS)) or percentage of energy from fat (F = 0.7, NS) as a result of the intervention. No intervention effects were found in boys or in girls for fruit (F = 0.5, NS), soft drinks (F = 2.6, NS) and water consumption (F = 0.3, NS). CONCLUSIONS: Combining physical and social environmental changes with computer-tailored feedback in girls and their parents can induce lower fat intake in middle-school girls. However, to have an impact on the consumption of soft drinks and water, governmental laws that restrict the at school availability of low-nutritive products may be necessary. PMID- 17411464 TI - Overweight and obesity among Iranian female adolescents in Rasht: more overweight in the lower social group. AB - OBJECTIVE: This study aimed to investigate the frequency of overweight and obesity among Iranian adolescent girls aged 14-17 years and its possible association with maternal education in Rasht city in 2005. DESIGN: A cross sectional survey on 1054 schoolgirls. SETTING: High schools in Rasht city, Iran. METHODS: A multistage sampling method was used and 1054 randomly selected schoolgirls were studied. Data on age and mother's years of schooling were collected by questionnaire and body weight and height were measured. Overweight and obesity were defined using age- and sex-specific body mass index (BMI) cut off points proposed by the International Obesity Task Force (IOTF), and compared with 85th and 95th BMI percentile curves of the IOTF reference population, respectively. Data analyses included two groups based on mother's years of schooling: less educated ( < 12 years) and more educated ( > or = 12 years). RESULTS: The overall prevalence of overweight and obesity in this population was 21.9 and 5.3%, respectively. Results of logistic regression analysis showed that the risk of overweight was higher for lower age group (odds ratio = 2.0, 95% confidence interval 0.18-3.09). Overweight was more common in girls with less educated mothers than in girls with more educated mothers (26.0 vs. 19.8%, P = 0.03). CONCLUSION: These data indicate that overweight is highly prevalent among adolescent girls, especially in lower social groups in Rasht, and the rate is exceeding those reported in other parts of the country. Preventive strategies need to be adopted to combat the epidemic of overweight and obesity in this population. PMID- 17411465 TI - Dietary transition stages based on eating patterns and diet quality among Haitians of Montreal, Canada. AB - OBJECTIVES: To identify dietary transition stages based on dietary patterns of adult Haitians having lived in Montreal for various lengths of time, and to assess associated dietary quality. SUBJECTS: One hundred and eighty-one Haitians aged 25-60 years were recruited by the modified random digit dialling method. DESIGN: Socio-economic, diet and lifestyle variables were documented by questionnaire. Three non-consecutive 24-hour dietary recalls were conducted over a 3-month period. Dietary patterns were studied using cluster analysis, and their association with proportion of lifetime in Canada and with socio-economic status (SES) was examined. Diet quality criteria were micronutrient adequacy and healthfulness based on dietary recommendations of the World Health Organization. RESULTS: Four dietary phenotypes were identified which could roughly represent stages of dietary transition or acculturation (Traditional, Pre-Western, Western and Modern). Subjects in the 'Traditional' cluster were older and had lived for a significantly lower proportion of their lifetime in Canada; they also tended to be of lower SES. Diet quality was significantly higher in the 'Traditional' than the 'Western' type, particularly with respect to healthfulness. A significantly lower proportion of subjects complying with limited intake of total fat ( < 30%) and cholesterol ( < 45%) was observed in the 'Western' compared with other diet phenotypes. Less than 15% of all subjects consumed enough dietary fibre, irrespective of diet type. CONCLUSION: Dietary transition stages could be identified on the basis of food patterns of Haitians according to the proportion of their lifetime in Canada. Encouraging the youth to retain the traditional food culture in its positive aspects would appear relevant PMID- 17411466 TI - Dietary intake and development of a quantitative food-frequency questionnaire for the Barbados National Cancer Study. AB - OBJECTIVE: To develop a quantitative food-frequency questionnaire (QFFQ) for the Barbados National Cancer Study (BNCS) that will permit examination of associations of diet with breast and prostate cancer. DESIGN: Population intake data from the year 2000 Barbados Food Consumption and Anthropometric Surveys (BFCAS 2000) were used to derive a list of foods consumed by the population. A 192-item draft QFFQ was developed from this list. SETTING: Barbados, West Indies provides an ideal environment to understand cancer risk in African-origin populations, with high relevance to African-Americans. The BNCS is a population based case-control study examining risk factors for breast and prostate cancer in such populations. SUBJECTS: A total of 1600 persons, 18 years and older, completed a 24-hour recall in the BFCAS 2000. Fifty of 63 randomly selected residents (79% response rate) gave additional updated information on foods consumed. RESULTS: The 50 participants provided a one-time 24-hour recall and completed the draft QFFQ. The final instrument contains 148 items: breads, cakes, cereals = 17; rice, pastas, noodles = 8; dairy = 10; meat, fish, poultry = 42; fruit = 16; vegetables = 26; soft drinks = 14; alcoholic beverages = 5; others = 10. Additional questions include supplement use and food preparation methods such as grilling. CONCLUSION: The final instrument is concise, complete and the most up-to-date for assessing the food and nutrient intake of African-origin Barbadians and the associations with breast and prostate cancer. PMID- 17411467 TI - Periconceptional use of folic acid in Shanxi Province of northern China. AB - OBJECTIVE: To understand the periconceptional use of folic acid and its association with selected characteristics in northern China. DESIGN: As part of a population-based case-control study on external structural birth defects, we analysed periconceptional use of folic acid among control mothers. SETTING: The study was conducted in four counties (Pingding, Xiyang, Taigu and Zezhou) in Shanxi Province of northern China. SUBJECTS: The subjects in this study were the control mothers, who were interviewed between January 2003 and May 2005. RESULTS: Of the 480 respondent mothers, 49 women (10.2%) reported ever using folic acid supplements, and only 16 women (3.3%) used it periconceptionally. The rates of any use and periconceptional use did not change significantly with study year. Periconceptional use of folic acid was significantly increased among women with more than high school education (odds ratio (OR) = 4.57, P < 0.10), women who were not farmers (OR = 4.72, P < 0.05), women with a history of birth-defect affected pregnancy (OR = 32.73, P < 0.05) and women who planned their pregnancy (OR = 7.88, P < 0.10). CONCLUSION: The rate of periconceptional folic acid use was among the lowest of those reported from other countries. More work should be done to improve preconceptional use of folic acid with special attention on less educated women and women farmers. PMID- 17411468 TI - A comprehensive physical activity promotion programme at elementary school: the effects on physical activity, physical fitness and psychosocial correlates of physical activity. AB - OBJECTIVE: To evaluate the effects of a comprehensive physical activity (PA) promotion programme in elementary schools on children's total PA levels, leisure time PA, physical fitness and psychosocial correlates of PA. DESIGN: A pre-test post-test design over two school years. SETTING AND SUBJECTS: Sixteen elementary schools (764 children, mean age: 11.2 +/- 0.7 years) were randomly assigned to the intervention condition (n = 8) and the control condition (n = 8). The intervention included a health-related physical education programme, an extracurricular PA promotion programme and classroom-based PA education lessons. In the total sample, leisure-time PA, psychosocial correlates of PA and physical fitness were measured using a PA questionnaire and the Eurofit test battery. In a sub-sample, total PA levels were measured using an accelerometer. RESULTS: According to accelerometer data, children's moderate PA and moderate-to-vigorous PA (MVPA) levels decreased less in the intervention schools than in the control schools (P < 0.01). The average time spent on MVPA decreased by 9 min per day in the intervention schools compared with 33 min per day in the control schools. Children in the intervention schools reported significantly more moderate PA in leisure time than the controls (P < 0.05). No overall improvement of physical fitness and no effects on the psychosocial correlates of PA were found. CONCLUSIONS: The comprehensive PA promotion programme was successful in preventing a decline in children's total activity levels. Furthermore, the intervention increased children's PA engagement in leisure time. Therefore, implementation needs to be encouraged. PMID- 17411469 TI - Growth pattern and prevalence of obesity in affluent schoolchildren of Delhi. AB - OBJECTIVE: To investigate the prevalence of obesity in affluent schoolchildren of Delhi. DESIGN: This was a cross-sectional study covering over 4000 students. Anthropometric measurements and birth weights of all the students were recorded. SETTING AND SUBJECTS: The study was carried out in a school catering to the affluent section of Indian society and included both boys and girls in the age range 4 to 17 years. RESULTS: The heights and weights of the children were almost similar to those of the National Center for Health Statistics standards, and were much above the values reported by the Indian National Nutrition Monitoring Bureau, which largely captures rural children belonging to lower socio-economic strata in India. Of the subjects studied, 22% were overweight and 6% were obese. Only 6% of these children had low birth weight. CONCLUSION: The prevalence of obesity is rising among children because of their change in lifestyle. Children born with a birth weight of > or = 3 kg tended to have higher body mass index in their adolescent years and maybe consequently in their adult years. Nutrition education can play an important part in reducing the incidence of overweight/obesity and its associated complications. PMID- 17411470 TI - Plant breeding to control zinc deficiency in India: how cost-effective is biofortification? AB - OBJECTIVE: To estimate the potential impact of zinc biofortification of rice and wheat on public health in India and to evaluate its cost-effectiveness compared with alternative interventions and international standards. DESIGN: The burden of zinc deficiency (ZnD) in India was expressed in disability-adjusted life years (DALYs) lost. Current zinc intakes were derived from a nationally representative household food consumption survey (30-day recall) and attributed to household members based on adult equivalent weights. Using a dose-response function, projected increased zinc intakes from biofortified rice and wheat were translated into potential health improvements for pessimistic and optimistic scenarios. After estimating the costs of developing and disseminating the new varieties, the cost-effectiveness of zinc biofortification was calculated for both scenarios and compared with alternative micronutrient interventions and international reference standards. SETTING: India. SUBJECTS: Representative household survey (n = 119 554). RESULTS: The calculated annual burden of ZnD in India is 2.8 million DALYs lost. Zinc biofortification of rice and wheat may reduce this burden by 20-51% and save 0.6-1.4 million DALYs each year, depending on the scenario. The cost for saving one DALY amounts to $US 0.73-7.31, which is very cost-effective by standards of the World Bank and the World Health Organization, and is lower than that of most other micronutrient interventions. CONCLUSIONS: Not only may zinc biofortification save lives and prevent morbidity among millions of people, it may also help accommodate the need to economise and to allocate resources more efficiently. Further research is needed to corroborate these findings. PMID- 17411471 TI - The infant feeding practices of mothers in the Maldives. AB - OBJECTIVE: To document the infant feeding practices of Maldivian mothers. METHODS: A cross-sectional survey of 251 Maldivian mothers recruited in 2004 from the 'well baby' or 'well child' clinics on the island of Male' and including 75 mothers from three other islands, Hura, Himmafushi and Thulusdhoo. RESULTS: The full breast-feeding rate at hospital discharge was 93% but declined to 41% at 4 months. Any breast-feeding rates were high among Maldivian mothers: 100% at 1 month and 85% were still breast-feeding at 6 months. The median duration of breast-feeding was 24 months. CONCLUSION: Breast-feeding rates are high and the average duration of breast-feeding is more than 2 years in the Maldives. Health promotion activities should be directed towards maintaining the already high 'any breast-feeding' rates and increasing the proportion of infants exclusively breast fed to 6 months. PMID- 17411472 TI - Dependence of blood indices of selenium and mercury on estimated fish intake in a national survey of British adults. AB - OBJECTIVE: Contributions of fish and other foods to variance of selenium and mercury status were studied in British adults. SETTING AND DESIGN: Plasma and red cell selenium and whole-blood mercury concentrations were measured during the National Diet and Nutrition Survey of Adults aged 19-64 years in mainland Britain, 2000-2001 (n = 1216). Food intake was weighed for seven consecutive days, and foods were combined in groups for data analysis. Four subsidiary groups characterised fish intakes: fried white fish, 'other' white fish, shellfish and oily fish. RESULTS: Geometric means and 5-95% ranges were: for whole-blood mercury, 5.61 (1.30-22.2) nmol l(-1); for plasma selenium, 1.09 (0.83-1.43) micromol l(-1); for red-cell selenium, 1.64 (1.14-2.40) micromol l(-1). Twenty eight per cent had no fish intake recorded during 7 days; the remaining 72% had a median intake of 237 g over the 7-day period, 5-95% range 45-780 g. Total fish intake was strongly and directly correlated with blood mercury, and moderately with red-cell and plasma selenium. Thus, sqrt(total fish intake) was correlated with: loge(blood Hg), t = +19.7; loge(plasma Se), t = +9.8; and loge(red-cell Se), t = +9.6, all P < 0.0001. All three biochemical (mercury and selenium) indices were strongly correlated with oily fish intake, and moderately correlated with shellfish and 'other' ( = non-fried) white fish, but none was significantly correlated with fried white fish. Blood mercury was strongly and directly correlated with red-cell and plasma selenium, and both increased with age. CONCLUSIONS: Dietary fish, especially oily fish, is a strong predictor of blood mercury and selenium in British adults. PMID- 17411473 TI - Educational variations in the consumption of foods containing fat in Finland and the Baltic countries. AB - OBJECTIVE: To examine educational differences among people who consume foods containing fat in Finland and the Baltic countries. DESIGN: Data were collected from cross-sectional postal Finbalt Health Monitor surveys that were carried out in 1998, 2000 and 2002. SETTING: Estonia, Finland, Latvia and Lithuania. SUBJECTS: For each survey, nationally representative random samples of adults aged 20-64 years were drawn from population registers (Estonia, n = 3656; Finland, n = 9354, Latvia, n = 6015; Lithuania, n = 5944). RESULTS: Differences were revealed between the countries in the consumption of foods that contain fat. Finnish people consumed butter on bread, high-fat milk, meat and meat products, and vegetable oil for cooking less frequently than people in the Baltic countries. Cheese was most popular in Finland. Educational differences in fat related food habits were examined by applying logistic regression analysis. A positive association was found between level of education and consumption of vegetable oil used in food preparation. Drinking high-fat milk was associated with low education in all countries. People with higher education tended to consume cheese more often. Educational patterns in the consumption of butter on bread and in the consumption of meat and meat products differed between countries. CONCLUSIONS: The consumption of foods containing fat was related to educational levels in all four countries. The diet of better-educated people was closer to recommendations for the consumption of saturated and unsaturated fats than the diet of people with lower level of education. The educational gradient was more consistent in Finland than in the Baltic countries. These existing educational differences in sources of fat consumed should be taken into account in the development of national cardiovascular disease prevention programmes. PMID- 17411475 TI - Influence of temperature, photoperiod and humidity on oviposition and egg hatch of the root-feeding flea beetle Longitarsus bethae (Chrysomelidae: Alticinae), a natural enemy of the weed Lantana camara (Verbenaceae). AB - The root-feeding flea beetle Longitarsus bethae Savini & Escalona, was introduced into South Africa as a candidate biological control agent for the noxious and invasive weed, Lantana camara L. As part of the study to predict the beetles' survival in its new range, the influence of climatic conditions on its egg development and reproductive performance were investigated in the laboratory. The threshold temperature (T degrees) and degree-days (DD) required for egg hatch were determined after exposing the eggs to various constant temperatures (12, 17, 22, 27 and 32 degrees C) in separate growth chambers. The DD required for egg hatch was 178.6, and the temperature threshold required for egg hatch was 11.3 degrees C. Survival of eggs varied from 27 to 56% at 32 and 17 degrees C, respectively, and was optimum between 17 and 25 degrees C. Oviposition was examined under high and low relative humidity (RH) regimes while egg hatch was determined at six RH levels, each maintained in a separate controlled growth chamber set at a constant temperature (25 degrees C). Whilst RH had no influence on oviposition, eggs were highly susceptible to aridity, and continuous exposure to relative humidity below 63% for more than three days was wholly lethal at 25 degrees C. Optimum egg hatch occurred at RH between 85 and 95% for up to 12 days. The effect of day length on oviposition and subsequent egg hatch was investigated under two photoperiod regimes. Neither oviposition nor subsequent egg hatch was influenced by photoperiod. The knowledge obtained will be useful for mass rearing as well as field release programmes for L. bethae. PMID- 17411474 TI - Comparative validation of standard, picture-sort and meal-based food-frequency questionnaires adapted for an elderly population of low socio-economic status. AB - OBJECTIVE: To compare the validity of a modified Block food-frequency questionnaire (FFQ), a picture-sort administration of the FFQ (PSFFQ) and a meal pattern-based questionnaire (MPQ) in a multi-ethnic population of low socio economic status (SES). DESIGN: Participants completed six 24-hour dietary recalls (24HR) over six months; the FFQ, PSFFQ and MPQ were completed in random order in the subsequent month. Instruments were interviewer-administered. The PSFFQ and MPQ were developed in formative research concerning difficulties for older adults in responding to standard food-frequency instruments. SETTING: Rural North Carolina, USA. Subjects One hundred and twenty-two African American, Native American and white adults aged > or = 65 years, with approximately one-third in each ethnic group. Inclusion criteria included education < or = 12 years and income < or = 150% of national poverty level or Medicaid recipient. RESULTS: Comparing median intakes from the average of the 24HR with the three diet assessment instruments, the MPQ tended to overestimate intakes compared with the FFQ and PSFFQ. Correlations among nutrients obtained by the 24HR and the other three instruments were generally statistically significant and positive. Across nutrients, the PSFFQ was most highly correlated with the 24HR for women, while the FFQ was most highly correlated with the 24HR for men. CONCLUSIONS: Dietary assessments using 24HR and FFQ were similar to results reported elsewhere, although correlations between 24HR and FFQ were somewhat lower. Interviewer administered dietary assessments should be used with caution to evaluate dietary intake among older adults with low SES. Gender differences and the lower correlations should be investigated more thoroughly to assist in choosing dietary assessment instruments for this population. PMID- 17411476 TI - Habitat use and phenology of the large insular endemic grasshopper Acrostira euphorbiae (Orthoptera: Pamphagidae). AB - The habitat use and the phenology of the large grasshopper Acrostira euphorbiae Garcia and Oromi endemic to La Palma (Canary Islands) are studied. This grasshopper is entirely dependent on the Canarian endemic shrub Euphorbia lamarckii both for food and to avoid predation. Adults stay on subapical branches during the day, probably to reduce the risk of predation, and climb up to the apex at night to feed. While females seem to ensure the genetic diversity of offspring by waiting for visits by different males, the latter have to move to guarantee their reproductive success. Monophagy in this species may be related to the year-round presence of tender shoots, and to the predator-repellent toxic latex found in the host plant. Unlike related species from continental areas, adults and nymphs of A. euphorbiae are present almost all year round, probably in adaptation to the particular climate of the islands. Nymphs are more abundant in winter, when Euphorbia leaves are most available. However, adults are more abundant than nymphs in spring, summer and autumn. Males develop more quickly than females, an apparent reproductive strategy based on achieving sexual maturity to coincide with females undergoing imaginal moulting. Matings start immediately after adult females emerge. Densities oscillated between 73 and 193 individuals ha(-1), which can be considered a low value compared with other continental pamphagid species. PMID- 17411477 TI - Effects of avermectin residues in cattle dung on yellow dung fly Scathophaga stercoraria (Diptera: Scathophagidae) populations in grazed pastures. AB - The effects of avermectin exposure on natural populations of the yellow dung fly, Scathophaga stercoraria Linnaeus, were investigated at the field scale on farms in south-west Scotland. Pastures forming the focus of the study were grazed with either untreated cattle or cattle receiving standard, manufacturer-recommended treatment regimes of an avermectin product. Flies were sampled between April and July in 2002 and 2003 using dung-baited pitfall traps. Abundance and wing asymmetry in S. stercoraria populations were examined in relation to a range of environmental and management variables (including avermectin exposure, pasture management intensity, weather and season). Data used for abundance analyses were collected in fields where treated cattle had been dosed with either doramectin or ivermectin, while the data for the asymmetry analyses were from a subset of fields where treated cattle had been dosed with doramectin only. While abundance of S. stercoraria varied significantly between years and with season, there was no difference in their abundance between fields grazed by avermectin-treated or untreated cattle. Asymmetry was significantly higher in fly populations in fields grazed by doramectin-treated cattle, suggesting that exposure to doramectin during development could have imposed some degree of environmental stress. While these results suggest that exposure to doramectin residues in dung on grazed pastures may have sublethal effects on the insects developing in that dung, there was no evident avermectin effect on the abundance of adult S. stercoraria occurring in the pastures. PMID- 17411478 TI - A taxonomic revision, biology and morphology of immature stages of the Entedon sparetus species group (Hymenoptera: Eulophidae), egg-larval endoparasitoids of weevils (Coleoptera: Curculionidae). AB - The sparetus-group of Entedon is revised. Six species are recognized for the group, including one new species, E. lucasi Gumovsky sp. n., which is described from Algeria. All species are either diagnosed or described, and illustrated, and new distribution records are provided. An illustrated dichotomous key to all species of the group is given. Biology of two species, E. thomsonianus Erdos and E. sparetus Walker, is studied in detail: both species proved to be egg-larval parasitoids of stem-boring weevils, Lixus cardui Olivier on the scotch thistle Onopordum acanthium and Gymnetron asellus (Gravenhorst) on mullein Verbascum sp., respectively. The morphology of larval stages and peculiarities of parasitoid host relationships are described. Possible benefits of egg-larval parasitism for Entedon species are discussed. PMID- 17411479 TI - Genetic differentiation of the pine wilt disease vector Monochamus alternatus (Coleoptera: Cerambycidae) over a mountain range - revealed from microsatellite DNA markers. AB - To study the dispersal process of the pine sawyer Monochamus alternatus (Hope) in frontier populations, a microsatellite marker-based genetic analysis was performed on expanding populations at the northern limit of its range in Japan. In Asian countries, M. alternatus is the main vector of pine wilt disease, the most serious forest disease in Japan. Sawyers were collected from nine sites near the frontier of the pine wilt disease damage area. A mountain range divides the population into western and eastern sides. Five microsatellite loci were examined and a total of 188 individuals was genotyped from each locus with the number of alleles ranged from two to nine. The mean observed heterozygosity for all loci varied from 0.282 to 0.480 in the nine sites, with an overall mean of 0.364. None of the populations have experienced a significant bottleneck. Significant differentiation was found across the mountain range, but the genetic composition was similar amongst populations of each side. It is believed that the mountain range acts as a geographical barrier to dispersal and that gene flow without a geographical barrier is high. On the west side of the mountain range, a pattern of isolation by distance was detected. This was likely to be caused by secondary contact of different colonizing routes on a small spatial scale. Based on these data, a process linking genetic structure at local (kilometres) and regional spatial scales (hundreds of kilometres) was proposed. PMID- 17411480 TI - Water stress augments silicon-mediated resistance of susceptible sugarcane cultivars to the stalk borer Eldana saccharina (Lepidoptera: Pyralidae). AB - Silicon (Si) can improve resistance of plants to insect attack and may also enhance tolerance of water stress. This study tested if Si-mediated host plant resistance to insect attack was augmented by water stress. Four sugarcane cultivars, two resistant (N21, N33) and two susceptible (N26, N11) to Eldana saccharina Walker were grown in a pot trial in Si-deficient river sand, with (Si+) and without (Si-) calcium silicate. To induce water stress, irrigation to half the trial was reduced after 8.5 months. The trial was artificially infested with E. saccharina eggs after water reduction and harvested 66 days later. Silicon treated, stressed and non-stressed plants of the same cultivar did not differ appreciably in Si content. Decreases in numbers of borers recovered and stalk damage were not associated with comparable increases in rind hardness in Si+ cane, particularly in water-stressed susceptible cultivars. Overall, Si+ plants displayed increased resistance to E. saccharina attack compared with Si- plants. Borer recoveries were significantly lower in stressed Si+ cane compared with either stressed Si- or non-stressed Si- and Si+ cane. Generally, fewer borers were recovered from resistant cultivars than susceptible cultivars. Stalk damage was significantly lower in Si+ cane than in Si- cane, for N21, N11 and N26. Stalk damage was significantly less in Si+ combined susceptible cultivars than in Si- combined susceptible cultivars under non-stressed and especially stressed conditions. In general, the reduction in borer numbers and stalk damage in Si+ plants was greater for water-stressed cane than non-stressed cane, particularly for susceptible sugarcane cultivars. The hypothesis that Si affords greater protection against E. saccharina borer attack in water-stressed sugarcane than in non-stressed cane and that this benefit is greatly enhanced in susceptible cultivars is supported. A possible active role for soluble Si in defence against E. saccharina is proposed. PMID- 17411481 TI - Parasitism, development and adult longevity of the egg parasitoid Telenomus nawai (Hymenoptera: Scelionidae) on the eggs of Spodoptera litura (Lepidoptera: Noctuidae). AB - When Telenomus nawai Ashmead or Trichogramma ostriniae Pang & Chen (Hymenoptera: Trichogrammatidae) was inoculated into intact egg masses of Spodoptera litura (Fabricius) that were covered with a scale-hair layer, T. nawai emerged from 76% of the eggs, while emergence of T. ostriniae was from only 3% of the eggs. When the hair layer was removed before inoculation, the rate by the latter increased to 21%. These observations confirmed that the hair layer effectively protects S. litura egg masses from attack by T. ostriniae, and that T. nawai parasitism is more effective. In order to assess the feasibility of T. nawai as a biological control agent for S. litura, parasitism rate, development time and adult longevity were examined in the laboratory. Emergence of T. nawai was observed in more than 95% of 0- and 1-day-old separated eggs of S. litura, but the rate decreased to 60% and 0% for 2- and 3-day-old eggs, respectively. The emergence rates of T. nawai were near 95% for temperatures in a range from 25 degrees C to 35 degrees C, but decreased to 80% at 20 degrees C. No parasitoids emerged at 15 degrees C. The developmental periods decreased as temperature increased from 20 degrees C to 35 degrees C. The developmental threshold occurred at 13.7 degrees C and 13.9 degrees C, and the effective accumulative temperatures from egg to adult emergence were 149.3 and 147.1 degree-days for females and males, respectively. Mean longevity of the adult females decreased with increasing temperature; 87.0 days at 15 degrees C and 9.5 days at 35 degrees C. The feasibility of the use of T. nawai for controlling S. litura is discussed. PMID- 17411482 TI - Evidence for resistance to pyrethroids and organophosphates in Plutella xylostella (Lepidoptera: Plutellidae) from Pakistan. AB - The susceptibility of representative pyrethroid (cypermethrin, deltamethrin, lambdacyhalothrin, bifenthrin), organophosphate (chlorpyriphos, triazophos, profenophos) and new chemistry insecticides (spinosad, indoxacarb and emamectin) was investigated for 18 field populations of Plutella xylostella (Linnaeus) from three different zones in Pakistan. The LC(50) (mg ml(-1); 48 h) values of pyrethroids for various populations ranged from 0.19-1.88 for cypermethrin, 0.31 2.64 for deltamethrin, 0.08-1.16 for lambdacyhalothrin and 0.07-0.88 for bifenthrin. The LC(50) (mg ml(-1); 48 h) of organophosphates ranged from 0.52 5.67 for chlorpyriphos, 0.37-4.14 for triazophos and 0.03-2.65 for profenophos. The most probable reason for low toxicity of organophosphates and pyrethroids is the evolution of multiple resistance mechanisms; however, further studies are required to establish these mechanisms. When these same products were tested against a susceptible laboratory population (Lab-Pak), the new chemistry compounds were significantly more toxic than pyrethroids and organophosphates. The results are discussed in relation to integrated pest management and insecticide resistance management strategies for P. xylostella. PMID- 17411483 TI - Feeding mode and prey detectability half-lives in molecular gut-content analysis: an example with two predators of the Colorado potato beetle. AB - The time during which prey remains are detectable in the gut of a predator is an important consideration in the interpretation of molecular gut-content data, because predators with longer detectability times may appear on the basis of unweighted data to be disproportionately important agents of prey population suppression. The rate of decay in detectability, typically expressed as the half life, depends on many variables; one that has not been explicitly examined is the manner in which the predator processes prey items. The influence of differences in feeding mode and digestive physiology on the half-life of DNA for a single prey species, the Colorado potato beetle Leptinotarsa decemlineata (Say), is examined in two predators that differ dramatically in these attributes: the pink ladybeetle, Coleomegilla maculata (DeGeer), which feeds by chewing and then ingesting the macerated material into the gut for digestion; and the spined soldier bug, Podisus maculiventris (Say), which physically and enzymatically processes the prey extra-orally before ingestion and further digestion in the gut. In order to standardize the amount of DNA consumed per predator, a single L. decemlineata egg was used as the prey item; all predators were third instars. The PCR assay yields estimated prey DNA half-lives, for animals maintained under field temperatures, of 7.0 h in C. maculata and 50.9 h in P. maculiventris. The difference in the prey DNA half-lives from these two predators underscores the need to determine detectabilities from assemblages of predators differing in feeding mode and digestive physiology, in order to weight positives properly, and hence determine the predators' relative impacts on prey population suppression. PMID- 17411484 TI - Evaluation of Bt-toxin uptake by the non-target herbivore, Myzus persicae (Hemiptera: Aphididae), feeding on transgenic oilseed rape. AB - As consequence of the concern about the biosafety of genetically modified plants, biological and ecological studies are considered crucial for environmental risk assessment. Laboratory experiments were carried out in order to evaluate the transfer of the Cry1Ac Bt-toxin from a transgenic Bt-oilseed rape to a non-target pest, Myzus persicae Sulzer. Cry1Ac protein levels in plants and aphids were determined using a double sandwich enzyme-linked immunosorbent assay. Phloem sap from (Bt+) and (Bt-) oilseed rape plants was collected from leaves using a standard method of extraction in an EDTA buffer. Bt-toxin was present in phloem sap, with a mean concentration of 2.7 +/- 1.46 ppb, corresponding to a 24-fold lower level than in oilseed rape leaves. Toxin was also detected in aphid samples, with a mean concentration in the positive samples of 2.0 +/- 0.8 ppb. The evidence that Bt-toxin remains in herbivores, in this case an aphid, could be useful to clarify functional aspects linked to possible consequences of Bt-crops on food chains involving herbivore-natural enemy trophic systems. Further studies are needed in order to improve the knowledge on the functional aspects linked to the transfer of the Cry1Ac Bt-toxin from GM-oilseed rape to aphids and their possible consequence. PMID- 17411485 TI - Educational psychology and the effectiveness of inclusive education/mainstreaming. AB - BACKGROUND: Inclusive education/mainstreaming is a key policy objective for the education of children and young people with special educational needs (SEN) and disabilities. AIMS: This paper reviews the literature on the effectiveness of inclusive education/mainstreaming. The focus is on evidence for effects in terms of child outcomes with examination also of evidence on processes that support effectiveness. SAMPLES: The review covers a range of SEN and children from pre school to the end of compulsory education. METHOD: Following an historical review of evidence on inclusive education/mainstreaming, the core of the paper is a detailed examination of all the papers published in eight journals from the field of special education published 2001-2005 (N=1373): Journal of Special Education, Exceptional Children, Learning Disabilities Research and Practice, Journal of Learning Disabilities, Remedial and Special Education, British Journal of Special Education, European Journal of Special Needs Education, and the International Journal of Inclusive Education. The derived categories were: comparative studies of outcomes: other outcome studies; non-comparative qualitative studies including non-experimental case studies; teacher practice and development; teacher attitudes; and the use of teaching assistants. RESULTS: Only 14 papers (1.0%) were identified as comparative outcome studies of children with some form of SEN. Measures used varied but included social as well as educational outcomes. Other papers included qualitative studies of inclusive practice, some of which used a non-comparative case study design while others were based on respondent's judgements, or explored process factors including teacher attitudes and the use of teaching assistants. CONCLUSIONS: Inclusive education/mainstreaming has been promoted on two bases: the rights of children to be included in mainstream education and the proposition that inclusive education is more effective. This review focuses on the latter issue. The evidence from this review does not provide a clear endorsement for the positive effects of inclusion. There is a lack of evidence from appropriate studies and, where evidence does exist, the balance was only marginally positive. It is argued that the policy has been driven by a concern for children's rights. The important task now is to research more thoroughly the mediators and moderators that support the optimal education for children with SEN and disabilities and, as a consequence, develop an evidence based approach to these children's education. PMID- 17411486 TI - Thirty years on - a large anti-Flynn effect? The Piagetian test Volume & Heaviness norms 1975-2003. AB - BACKGROUND: Volume & Heaviness was one of three Piagetian tests used in the CSMS survey in 1975/76. However unlike psychometric tests showing the Flynn effect - that is with students showing steady improvements year by year requiring tests to be restandardized - it appeared that the performance of Y7 students has recently been getting steadily worse. AIMS: A sample of schools sufficiently large and representative was chosen so that the hypothesis of worsening performance could be tested, and estimated quantitatively. SAMPLE: Sixty-nine Y7 school year groups containing pupil data on the Volume & Heaviness test and the University of Durham CEM Centre MidYIS test were located giving a sample of 10, 023 students covering the years 2000 to 2003. METHOD: Regression of the students' school mean on Volume & Heaviness on the schools' mean MidYIS 1999 standardized score, and computing the regression at MidYS = 100 allows comparison with that found in 1976. RESULTS: The mean drops in scores from 1976 to 2003 were boys = 1.13 and girls = 0.6 levels. A differential of 0.50 standard deviations in favour of boys in 1976 had completely disappeared by the year 2002. Between 1976 and 2003 the effect-size of the drop in the boys' performance was 1.04 standard deviations, and for girls was 0.55 standard deviations. CONCLUSION: The idea that children leaving primary school are getting more and more intelligent and competent - whether it is viewed in terms of the Flynn effect, or in terms of government statistics on performance in Key Stage 2 SATS in mathematics and science - is put into question by these findings. PMID- 17411487 TI - Effects of prior assistance in using analogies on young children's unprompted analogical problem solving over time: a microgenetic study. AB - BACKGROUND: Various studies on analogical problem solving have shown that children can be taught to use analogies within a single session, but it is not known whether they can be taught a strategy for using analogical problem solving that persists over a period of time. AIM: Our study focused on the effects of prior assistance in analogy use on children's unprompted analogical problem solving over time. SAMPLE: Participants were 144, 5- to 7-year-old children from kindergarten and grade 1. METHODS: A four-session microgenetic procedure was used. A number of children were given assistance in using analogies 1 week before the experiment started. Their analogy use over time was examined and compared with peers without such experiences. RESULTS AND CONCLUSIONS: The results indicate that prior assistance in using analogies has a beneficial effect, which may last for several weeks, on analogical problem solving in children aged 6-7 years or even younger. The study also showed the use of analogies in the absence of instructions, and variability in task behaviour in all groups. Since these findings may have implications for instructional practice, the authors recommend a novel study in an educational environment and with tasks from a more educational domain. PMID- 17411488 TI - Arithmetic problems at school: when there is an apparent contradiction between the situation model and the problem model. AB - BACKGROUND: Understanding and solving problems involves different levels of representation. On the one hand, there are logico-mathematical representations, or problem models (PMs), which contain information such as 'the size of the flock changed from 31 sheep to 42' while, on the other hand, there are more qualitative representations, or qualitative situation models (QSMs), comprising events expressed in everyday terms, such as 'a lot of lambs had been born'. AIMS: We set out to show that an apparent contradiction between a PM and a QSM can result in poorer performances, especially when the activation of formal representations is not fully automatic. SAMPLE: We tested 44 third years (mean age 8;6) and 46 fourth years (mean age 9; 8) from two primary schools in France. METHOD: Change problems and compare problems were drafted in two versions: one where the QSM and PM were consistent and one where they appeared to be contradictory but were not (referred to hereafter as the inconsistent version). For example, if the numerical data proved that the size of a flock of sheep had increased (PM), the consistent version would state that a lot of lambs had been born (QSM), whereas the inconsistent version would state that the wolf had devoured some of the sheep (QSM). Each pupil was given 7 consistent problems and 7 'inconsistent' ones to solve. RESULTS: For the change problems, errors were more frequent on the inconsistent versions, especially among the younger subjects. For the compare problems, there were more errors on the inconsistent versions regardless of the subjects' age. CONCLUSION: It is in situations where the problem schema is not automatically activated (younger subjects or compare problems) that the QSM plays its role of intermediate representation and causes performances to deteriorate if it does not correspond to the PM. PMID- 17411489 TI - Self-handicapping in school physical education: The influence of the motivational climate. AB - BACKGROUND: Self-handicapping is an attribution-related process whereby individuals create performance impediments/excuses to protect self-worth in socially evaluative environments. Thus, the prevailing motivational climate would appear to be an important factor when attempting to understand the situational self-handicapping process within school physical education. AIMS: Drawing from achievement goal theory, the study examined the effect of experimentally induced conditions (viz. task vs. ego) on situational self-handicapping. SAMPLE: Seventy British secondary school students (36 females and 34 males; M age = 11.98; SD=0.31). METHOD: Participants were randomly assigned to partake in a running endurance task in either an ego-involving (20 male students and 16 female students) or a task-involving (14 male students and 20 female students) condition. Prior to completing the experimental task, participants were given the opportunity to claim situational self-handicaps. Data for goal orientations, subjective climate perceptions, perceived ability and perceived task importance were also obtained. RESULTS: After determining the effectiveness of the experimental manipulation, results revealed participants in the ego-involving condition to report significantly more situational self-handicapping claims. Further, and after controlling for individual difference variables, the results of moderated hierarchical regression analysis revealed subjective perceptions of an ego-involving climate to be the main positive predictor of situational self handicapping. Although a weaker contributor to the percentage of variance explained, task orientation emerged as a negative predictor of situational self handicapping. CONCLUSIONS: The findings suggest that PE teachers would be prudent to minimize ego-involving situations should they wish to reduce situational self handicapping. PMID- 17411490 TI - Academic self-handicapping: the role of self-concept clarity and students' learning strategies. AB - BACKGROUND: Self-handicapping is linked to students' personal motivations, classroom goal structure, academic outcomes, global self-esteem and certainty of self-esteem. Academic self-handicapping has yet to be studied with respect to students' consistency in self-description and their description of themselves as learners. AIMS: This study examined students' self-esteem and self-concept clarity as well as their tendencies to employ deep- or surface-learning approaches and self-regulate while learning in relation to their self handicapping tendencies and exam performance. SAMPLE: Participants were 161 male and female Canadian, first-year university students. METHOD: Participants completed a series of questionnaires that measured their self-esteem, self concept clarity, approaches to learning, self-regulation and reflections on performance prior to and following their exam. RESULTS: Self-handicapping was negatively correlated with self-concept clarity, deep learning, self-regulated learning and exam grades, and positively correlated with surface learning and test anxiety. Regression analyses showed that self-concept clarity, self regulation, surface-learning and test anxiety scores predicted self-handicapping scores. Self-concept clarity, test anxiety scores, academic self-efficacy and self-regulation were predictors of mid-term exam grades. CONCLUSIONS: This study showed that students' self-concept clarity and learning strategies are related to their tendencies to self-handicap and their exam performance. The role of students' ways of learning and their self-concept clarity in self-handicapping and academic performance was explored. PMID- 17411491 TI - The influence of achievement goals on the constructive activity of low achievers during collaborative problem solving. AB - BACKGROUND: Previous research on small-group learning has found that level of constructive activity (solving or explaining how to solve problems using ideas stated or implied in the explanation provided by a partner) was a better predictor of post-test achievement than either a student's prior achievement or the quality of help received (Webb, Troper, & Fall, 1995). AIMS: The purpose of this study was to extend this research by examining the influence of additional factors, in particular, achievement goals and comprehension monitoring, on low achieving students' constructive activity after receiving help from a high achieving peer. SAMPLE: Thirty-two low achieving upper elementary students from an urban school district in the mid-west of the United States were paired with high achieving partners. METHODS: Videotape data from a previously reported study on peer collaboration were transcribed and reanalyzed. In that study, dyads were randomly assigned instructions designed to induce either a learning or performance goal and were videotaped as they worked together to solve a set of mathematical word problems. The following day, students were individually post tested on problems similar to the ones worked on in pairs. RESULTS: Consistent with previous research, low achieving students' level of constructive activity predicted post-test performance. In addition, constructive activity was found to mediate the relationship between achievement goals and learning. However, achievement goals were not related to low achievers constructive use of help. Instead, achievement goals were related to low achievers' relative accuracy in comprehension monitoring, which in turn was related to level of constructive activity. CONCLUSIONS: The meaning of these results for understanding the processes by which low achievers learn from peer help and implications for classroom practice are discussed. PMID- 17411492 TI - The effects of Peer Mediation with Young Children (PMYC) on children's cognitive modifiability. AB - Peer mediation with young children is a relatively novel approach aimed at teaching young children how to mediate to their peers. The main benefits of peer mediation are in developing children's mediation teaching style and cognitive modifiability. The peer mediation developed recently is based on Vygotsky's sociocultural and Feuerstein's mediated learning experience theories. The main objectives of the study were to investigate the effects of the Peer Mediation with Young Children (PMYC) programme on children's cognitive modifiability of mediators and learners and to study the effects of cognitive level of the learner and mediator on their cognitive modifiability following the programme. A sample of 178 pupils (89 mediators in Grade 3 and 89 learners in Grade 1) was randomly assigned to experimental (N=43 dyads) and control (N=46 dyads) groups. The mediators in the experimental group participated in the PMYC programme, whereas the mediators in the control group received a substitute intervention aimed at emphasizing general conditions of peer interaction. Following the intervention, mediators of both groups received a demonstration of a multimedia programme as a preparation for the peer mediation interaction and later taught it to their young counterparts. Following the teaching session (e.g. teaching of seriation problems), mediators in both groups were given a dynamic assessment measure of analogies. The learners however were given a test of seriation before and after the intervention. The findings showed that following the intervention the experimental mediators showed higher level of analogies scores, as well as higher improvement on the dynamic analogies measure as compared with control mediators. The experimental learners showed higher pre- to post-intervention achievements on the seriation problems as compared with control learners. The findings showed also that when there was a match between the mediator and learner's cognitive level (i.e. low-low or high-high) the differences between the experimental and control groups were minor and negligible. However, when the mediator-learner cognitive levels did not match, the experimental learners received higher scores than the control learners. Mediators in the experimental group had to cope with the incongruent cognitive level by facilitating their mediational approach and consequently enhancing the learner's performance. The findings are discussed in regard to Piaget, Vygotsky and Feuerstein's theories and recent research on peer assisted learning. PMID- 17411493 TI - The validity of physical aggression in predicting adolescent academic performance. AB - BACKGROUND: Aggression has a long history in academic research as both a criterion and a predictor variable and it is well documented that aggression is related to a variety of poor academic outcomes such as: lowered academic performance, absenteeism and lower graduation rates. However, recent research has implicated physical aggression as being predictive of lower academic performance. AIMS: The purpose of this study was to examine the role of the 'Big Five' personality traits of agreeableness, openness to experience, conscientiousness, neuroticism and extraversion and physical aggression in predicting the grade point averages (GPA) of adolescent students and to investigate whether or not there were differences in these relationships between male and female students. SAMPLE: A sample of 992 students in grades 9 to 12 from a high school in south eastern USA as part of a larger study examining the students' preparation for entry into the workforce. METHOD: The study was correlational in nature: students completed a personality inventory developed by the second author with the GPA information supplied by the school. RESULTS: Results indicated that physical aggression accounts for 16% of variance in GPA and it adds 7% to the prediction of GPA beyond the Big Five. The Big Five traits added only 1.5% to the prediction of GPA after controlling for physical aggression. Interestingly, a significantly larger amount of variance in GPA was predicted by physical aggression for females than for males. CONCLUSIONS: Aggression accounts for significantly more variance in the GPA of females than for males, even when controlling for the Big Five personality factors. Future research should examine the differences in the expression of aggression in males and females, as well as how this is affecting interactions between peers and between students and their teachers. PMID- 17411494 TI - Using a multitrait-multimethod analysis to examine conceptual similarities of three self-regulated learning inventories. AB - BACKGROUND: A programme of construct validity research is necessary to clarify previous research on self-regulation and to provide a stronger basis for future research. AIM: A multitrait-multimethod (MTMM) analysis was conducted to assess convergent and discriminant validity of three self-regulation measures: the Learning and Study Strategies Inventory (LASSI; Weinstein, 1987), the Motivated Strategies for Learning Questionnaire (MSLQ; Pintrich, Smith, Garcia, & McKeachie, 1993) and the Meta-cognitive Awareness Inventory (MAI; Schraw & Dennison, 1994). Method bias across all three inventories was also examined. SAMPLE AND METHOD: Three hundred and eighteen undergraduate university students (255 female, 61 male, 2 did not specify) were recruited from various courses to participate in research on perceptions about studying and study methods. Participants spent 30-60 minutes completing all three inventories. RESULTS: Evidence for convergent validity was found at the matrix level, but was attenuated when examined at the individual parameter level. Evidence for discriminant validity among traits was modest, and common method bias was evident across all three measures. CONCLUSIONS: Results revealed the three inventories yielded different results, which suggests that researchers should be selective in the inventory they use to assess self-regulated learning (SRL). PMID- 17411495 TI - Self-construal and social comparison effects. AB - BACKGROUND: Social comparison research usually demonstrates that students will have higher self-evaluation in downward comparison but lower self-evaluation in upward comparison. However, the existence of this contrast effect may depend on people's self-construal. The contrast effect may exist only for people with independent self-construal. For people with interdependent self-construal, the contrast effect may be attenuated. AIM: The study investigated the role of self construal as a moderator of the social comparison effects in authentic classrooms. SAMPLE: The participants were 96 Chinese seventh-grade students (41 male, 51 female and 4 unreported) from a secondary school in Hong Kong. METHOD: The experiment employed a 2 x 2 between-subjects design based on 2 levels of self construal (independent, interdependent) and 2 levels of comparison standard (upward comparison, downward comparison). The dependent variable was students' self-evaluation. RESULTS: A two-way ANOVA indicated a significant interaction between self-construal and comparison standard on self-evaluation. When the students' independent self-construal was activated, they reported higher self evaluation in downward comparison but lower self-evaluation in upward comparison. However, such a contrast effect was attenuated when the students' interdependent self-construal was activated. They reported high self-evaluation in both upward and downward comparisons. CONCLUSIONS: The outcome of social comparison depends on whether independent or interdependent self-construal is salient in the classroom. PMID- 17411496 TI - The association between job strain and emotional exhaustion in a cohort of 1,028 Finnish teachers. AB - BACKGROUND: Teachers' work overload has been the subject of intense research, and the results of these studies show that a substantial proportion of teachers perceive their job as very stressful. AIMS: To investigate how different formulations of high demands and low decision latitude was related to teachers' burnout, and to estimate the possible interaction between these factors. SAMPLE: The sample consisted of 1,028 school teachers. METHOD: Multivariate covariant analyses (MANCOVA) was used to evaluate the relationship between a high-strain job defined by 3 different cut-off points and burnout. Logistical regression analysis was used to estimate the separate and joint effects of demand and decision authority on emotional exhaustion. Interaction between high demands and low decision authority was analysed using relative excess risk due to interaction (RERI). An attributable proportion (AP) was calculated in order to estimate the proportion of emotionally exhausted teachers among those exposed to both risk factors that was attributable to their synergistic interaction. The group of teachers who perceived their job as a low-strain job was used as the reference group in the analysis. RESULTS: The effect of job strain on burnout was proved to be consistent and robust across alternative formulations. The main effect of high demands exceeded that of low decision authority in relation to emotional exhaustion. Furthermore, the 2 factors acted synergistically to increase the risk of burnout. CONCLUSIONS: In the case of burnout, teachers who perceived their job as highly demanding and low in control, 69% of the effect could be attributed to the synergism of these 2 factors. PMID- 17411497 TI - Job stressors, personality and burnout in primary school teachers. AB - BACKGROUND: Teaching is considered a highly stressful occupation. Burnout is a negative affective response occurring as a result of chronic work stress. While the early theories of burnout focused exclusively on work-related stressors, recent research adopts a more integrative approach where both environmental and individual factors are studied. Nevertheless, such studies are scarce with teacher samples. AIMS: The present cross-sectional study sought to investigate the association between burnout, personality characteristics and job stressors in primary school teachers from Cyprus. The study also investigates the relative contribution of these variables on the three facets of burnout - emotional exhaustion, depersonalization and reduced personal accomplishment. SAMPLE: A representative sample of 447 primary school teachers participated in the study. METHOD: Teachers completed measures of burnout, personality and job stressors along with demographic and professional data. Surveys were delivered by courier to schools, and were distributed at faculty meetings. RESULTS: Results showed that both personality and work-related stressors were associated with burnout dimensions. Neuroticism was a common predictor of all dimensions of burnout although in personal accomplishment had a different direction. Managing student misbehaviour and time constraints were found to systematically predict dimensions of burnout. CONCLUSIONS: Teachers' individual characteristics as well as job related stressors should be taken into consideration when studying the burnout phenomenon. The fact that each dimension of the syndrome is predicted by different variables should not remain unnoticed especially when designing and implementing intervention programmes to reduce burnout in teachers. PMID- 17411500 TI - UK Health Services Research Network: at last, a health services research organization. PMID- 17411501 TI - A culture of safety or coping? Ritualistic behaviours in the operating theatre. AB - OBJECTIVES: The creation of a 'safety culture' is a health services priority, yet there is little contemporary research examining the tacit, customary practices that relate to clinical risk. This paper investigates how the ritualistic behaviours of surgeons and anaesthetists serve to normalize risks within the operating theatre, thereby inhibiting organizational learning and enabling such risks to recur. METHODS: A two-year ethnographic study in the operating department of a large teaching hospital in the north of England, including observations of the organizational and clinical setting and interviews with 80 members of staff. RESULTS: Three ritualistic types of behaviour are identified and described. Each illustrates the taken-for-granted assumptions associated with clinical risk in the operating theatre and is characterized by a patterned response to risk, the first being to tolerate and endure risk, the second being to accommodate risk through slight modifications to clinical practice and the third being to innovate or implement unorthodox practices to control for risk. CONCLUSION: These ritualistic behaviours normalize risk within the operating theatre leading to the possibility that some threats will escape appropriate attention and may lead to patient harm. These culturally scripted behaviours also encourage a short-term reactive response to risk that emphasizes the importance of individual coping rather than the more systemic forms of learning associated with participation in incident reporting. This research extends and elaborates upon the current policy orthodoxy to better understand the cultural context of patient safety. PMID- 17411502 TI - Socioeconomic inequality in small area use of elective total hip replacement in the English National Health Service in 1991 and 2001. AB - OBJECTIVES: To compare socioeconomic inequality in small area use of elective total hip replacement in the English National Health Service (NHS) in 1991 and 2001. METHODS: Hospital Episode Statistics and Census data were aggregated to a common geography of 'frozen' 1991 English electoral wards. The Townsend deprivation score was used as the primary indicator of socioeconomic status for each ward, and the sensitivity analysis used other Census indicators. Two main measures of inequality were examined: the indirectly age-sex standardized utilization rate ratio between most and least deprived quintile groups, and the concentration index of deprivation-related inequality in age-sex standardized utilization ratios between small areas. Each standardized utilization ratio is the observed use divided by the expected use, if each age and sex group in the study population had the same use rate as the national population. RESULTS: In both years, observed use was below expected use for the bottom third of areas by socioeconomic status. The standardized utilization rate ratio between top and bottom Townsend quintiles fell from 1.41 (95% confidence interval [CI] 1.36-1.47) in 1991 to 1.27 (95% CI 1.23-1.32) in 2001. The proportionate increase in use required to bring the bottom quintile to the level of top thus fell significantly from 41% to 27%. The Town-send-based concentration index also fell from 0.069 (95% CI 0.059-0.079) in 1991 to 0.060 (95% CI 0.050-0.071) in 2001, although this fall was not statistically significant (P = 0.085). Other socioeconomic indicators yielded a similar pattern. CONCLUSIONS: Socioeconomic small area inequality in use of total hip replacement appears to have fallen between 1991 and 2001. One possible explanation is that increased hip replacement rates in the 1990s may have lowered barriers to access, thus allowing this health technology to diffuse further among lower socioeconomic groups. PMID- 17411503 TI - Retention and future job intentions of graduate primary care mental health workers: a newly developed role in England. AB - OBJECTIVES: Modernization of the English National Health Service (NHS) workforce has led to the introduction of a number of new roles. One such role is the graduate primary care mental health worker (PCMHW). Although generally successful in expanding primary mental health care, several challenges were raised by the initial implementation, including retention and development of effective career pathways for these workers. The aim of this study was to examine retention and future job intentions of graduate PCMHWs at the end of their one-year training. METHODS: Mail surveys of the first cohort of graduate PCMHWs recruited in 2004 were conducted at the beginning and end of their 12-month training. RESULTS: Satisfaction with training courses was generally low. Over half indicated they were likely to leave their graduate PCMHW post within the next few months. However, most intended to stay within the NHS. CONCLUSIONS: After training, problems were highlighted in relation to the quality of the training programmes and lack of development of an effective career pathway. This has serious consequences for the viability of this initiative. PMID- 17411504 TI - Describing the impact of health services and policy research. AB - OBJECTIVES: In an essentially applied area of research, there are particular pressures on health services and policy researchers to describe the impact of their work. However, specialized research impact assessments often require skills and resources beyond those available to individual researchers, and ad hoc accounts impose a considerable burden to generate. Further, these idiosyncratic accounts may not facilitate comparative analysis to inform research management, practice and assessment. This paper describes an initial attempt to develop a methodical approach to identify and describe research impact. METHODS: A Research Impact Framework was developed, drawing on the literature and interviews with researchers at the London School of Hygiene and Tropical Medicine, and was used to structure impact narratives of selected research projects. These narratives were based on semi-structured interviews with principal investigators and documentary analysis of the projects. RESULTS: Using the framework as a guide, researchers were relatively easily and methodically able to identify and present impacts of their work. Researchers' narratives contained verifiable evidence and highlighted a wide range of areas in which health services and policy research has impact. The standardized structure of the narratives also facilitated analysis across projects. Factors thought to positively influence the impact of research included researchers' continued involvement in research and policy networks, established track records in the field, and the ability to identify and use key influencing events, such as 'policy windows'. CONCLUSIONS: The framework helped develop research impact narratives and facilitated comparisons across projects, highlighting issues for research management and assessment. PMID- 17411505 TI - 'Should I stay or should I go?' Deciding whether to go to hospital after a 999 call. AB - OBJECTIVE: In most UK ambulance services, crews attending someone who has phoned the emergency services on '999' will take the patient to hospital, unless the patient makes the decision to stay at home (or wherever they happen to be when the ambulance arrives). Safety concerns have been raised about non-conveyance decisions. We undertook a study of one UK Ambulance Service to examine ambulance crew members' views on how decision-making about non-conveyance works in practice in relation to non-urgent calls. METHODS: A total of 25 paramedics took part in three focus groups. Focus groups were transcribed and analysed thematically. RESULTS: The ambulance service's apparently straightforward guidance on decision making about non-conveyance proved tricky in the messiness of the real world, for two reasons. The first was to do with the notion of the patient's capacity to make decisions and how this was interpreted. The second was to do with the complexity of the decision-making process, in which the patient, the crew and, in many cases, family or carers often take part in negotiation and de facto joint decision-making. CONCLUSIONS: There is a mismatch between policy and practice in relation to non-conveyance decisions. Findings should be built into research and service development in this rapidly changing field of practice in emergency and/or unscheduled care. The commonly accepted perspective on shared decision making should be extended to include the context of '999' ambulance calls. PMID- 17411506 TI - Patients' experiences of Western-style acupuncture: the influence of acupuncture 'dose', self-care strategies and integration. AB - OBJECTIVES: To investigate patients' perspectives of the process and outcome of Western-style acupuncture for chronic health problems. To use these results to inform the provision of acupuncture in health services in the UK. METHODS: A purposive sample of 18 patients who were having Western-style acupuncture, for the first time, for a health problem of at least three months duration, were interviewed twice over a four-month period using semi-structured interviews. Using a constant comparative method, the data were analysed across cases and within cases. RESULTS: The interviewees complained of chronic pain and moderate or severe disability which was resistant to conventional treatment. Their experience of acupuncture was diverse and varied according to the 'dosage' of acupuncture received, the inclusion of self-care strategies, and their relationship with the practitioner. These three factors were interlinked and constituted individual styles of practice for each practitioner. The majority of patients benefited in terms of complete or partial relief of pain and disability, and reduction in conventional medication. However, some patients were disappointed by the treatment, distressed about 'wasting people's time', and about the lack of continuity of care. People who benefited most had good general health and a single problem. CONCLUSIONS: Patients showed discerning judgement regarding the 'dosage' of acupuncture they required, and combined acupuncture with exercises to good effect. Publicly funded health services should provide an acupuncture service that provides the optimal 'dosage' and uses pain relief to promote self-care. Further research to investigate the benefits of a service that combines Western-style and traditional acupuncture is planned. PMID- 17411507 TI - Tied to the job: affective and relational components of nurse retention. AB - OBJECTIVE: To investigate whether affective and relational components of nurses' experience of work have a significant impact on their intentions to leave either the job or the nursing profession in models that control for other factors (sociodemographic, work conditions, perceptions of quality of care) that are known to affect career decisions. METHOD: An exploratory, cross-sectional postal survey of 2880 nurses in grades A-I in 20 National Health Service (NHS) Hospital Trusts, 11 in inner London and nine in outer London, was carried out between January and July 2002, looking at nurses' intention to leave their current job or the nursing profession. The data were analysed using logistic regression with robust standard errors. RESULTS: In models that controlled for known sources of job dissatisfaction, relationships with supervisors and managers were found to have a significant effect on respondents' career intentions. Feeling valued by the Trust and by society was very important. Nurses seemed to distinguish between local problems that are the responsibility of the Trust and those, such as levels of pay, that could only be solved at the national level. CONCLUSION: Nurses' career intentions are complex and multifactorial. Feelings of being valued and listened to play a role, as well as the individual and job-related characteristics. The study highlights the role of supervisors and managers in retaining staff and suggests that investment in robust systems of communication, conflict resolution and security could slow nurse turnover. The NHS as an employer may be most interested in the role of pay in nurse retention, and the general public in how societal attitudes and verbal abuse shape nurses' career decisions. PMID- 17411510 TI - Gotzsche's quixotic antiscreening campaign: nonscientific and contrary to Cochrane principles. PMID- 17411511 TI - The debate on breast cancer screening with mammography is important. PMID- 17411512 TI - Medical malpractice reform: federal politics. PMID- 17411513 TI - Research report from the NIH: introduction to the NIH and the NCI director's year 2015 challenge goal. PMID- 17411514 TI - The malpractice liability crisis. AB - Most medical malpractice cases are tried under the civil tort of negligence and are often triggered by adverse outcomes. These proceedings are aimed primarily at determining whether the conduct of a health care provider was reasonable. Such legal actions have mostly been subject to state jurisdiction. Increasingly, a number of factors are converging that are threatening the continued practice of medicine in some states and hence patients' access to care. These include higher amounts of monetary damages awarded to successful plaintiffs, consequent rising malpractice premiums, and the threatened economic insolvency of medical liability insurance carriers as a result of the broader economic downturn. The result is a serious public health dilemma. The national scope of the problem has been considered a crisis, which has prompted unprecedented federal legislative proposals directed toward providing new and preemptive parameters for capitated noneconomic damages, restrictions on certain civil procedures affecting lawsuit outcomes, and methods for attorney compensation, which some states have either not previously addressed or found unconstitutional. A survey of different states' problems and common issues should assist the reader in understanding the nature of the crisis and proposed solutions. PMID- 17411515 TI - Turf wars in radiology: introduction. AB - Turf wars over medical imaging have become ubiquitous. At many centers, radiologists already have lost access to important technologies and their applications. A combination of decreasing revenues in other specialties and a growing attachment to imaging further threatens radiologists' practices in the future. This is the first of an extended series of articles that will address the phenomenon of self-referral--its motivations, extent, and the threats it poses to radiologists. Each article will focus on a specific aspect and provide advice on how radiologists can deal with self-referral in their locales. PMID- 17411516 TI - Implementing an effective organization and governance structure for a radiology practice. AB - Radiology practices that are well organized and effectively governed have a competitive advantage. Decisions are made rapidly, actions are taken decisively and in accordance with established policy, and each group member has a responsibility for practice building. Such groups are perceived by their peers, hospital administration, and community business leaders to be both formidable and effective. This paper details the mechanisms that facilitate planning for and implementing an efficient practice organization and governance structure. The tasks of group leaders are defined, as are the committees necessary for appropriate action. The integral roles of a mission statement and a business plan are discussed. Practices adopting the suggested organizational structure will be best positioned to survive in both good times and bad. PMID- 17411517 TI - The future quality and safety of medical imaging: proceedings of the third annual ACR FORUM. AB - The ACR annually engages in a long-range planning activity called the FORUM on a specific issue deemed important to the specialty of radiology over a horizon of 5 to 10 years. The FORUM brings together experts from multiple disciplines to discuss the topic, develop scenarios, and make recommendations to the ACR and the specialty on what courses to take to improve the development of radiology. The Third Annual FORUM, held May 21 to 23, 2003, was on the subject of improving quality and safety. The sessions ended with numerous recommendations as to how the specialty of radiology could promote safer, higher quality imaging that would also engage the interest and participation of the public, employers, insurers, and government. These fell into the following categories: (1) how to measure the performance of radiologists, (2) how to use measurements of performance to improve quality and safety, (3) means of working with employers and insurers to improve care and reduce waste, (4) the development of new ACR programs intended to foster change, and (5) disseminating information about ACR efforts on behalf of improving quality and safety. PMID- 17411518 TI - Jung v. Association Of American Medical Colleges: the lawsuit challenging our system of graduate medical education. AB - This article examines the antitrust case in which three physicians have challenged the Match Program and various other aspects of the system of graduate medical education in the United States. After describing the parties to the litigation, the author explains the plaintiffs' theories of liability. He suggests how the plaintiffs are likely to claim damages in the multibillion dollar range and speculates about the injunctive relief that the plaintiffs are likely to seek. The author then sets forth some of the principal defenses to the case, as well as the basis for the defendants' position that the plaintiffs have not been injured at all. The author goes on to explore developments that have occurred since the lawsuit was filed on May 7, 2002. These include a motion by the National Residency Matching Program to refer the case to arbitration; motions by certain teaching hospital defendants asserting that they are not subject to suit in the District of Columbia; and motions by the American Medical Association, the Accreditation Council for Graduate Medical Education, and other organization defendants to the effect that the plaintiffs have not alleged conduct by them that violates the antitrust laws. The author then discusses the motion by the plaintiffs to have the case certified as a class action. Finally, he considers legislative initiatives that the litigation has engendered. PMID- 17411519 TI - From concept to CPT code to compensation: how the payment system works. AB - All radiologists and radiation oncologists provide medical services to patients every day with the full anticipation that these services will be appropriately reimbursed. Yet most take this process for granted. Few have even a rudimentary idea how the system works by which a coding mechanism and reimbursement schedule are developed and maintained for the vast array of services they provide. Clearly, this is not good business. You need not stay in the dark any longer! This article describes (1) the fundamental structure of reimbursement for radiology and radiation oncology services; (2) the multiple steps required as a new procedure advances from a research concept to the assignment of a code in the American Medical Association's Current Procedural Terminology; (3) the process by which the new procedure and code are assigned a reimbursement value in the Medicare Fee Schedule, which acts as the base for over 75% of current medical reimbursement; and (4) the maintenance of this system for existing procedures. PMID- 17411520 TI - Radiological and nuclear terrorism: are you prepared? AB - Another terrorist attack on our nation is virtually inevitable. Most believe that it is not a question of if but when. The form of the terrorism, the time, and the place will not be of our choosing. Radiology professionals (radiologists, technologists, radiologists' assistants, and nurses) will be involved in caring for the victims of the attack, whether the method employed is chemical, biological, radiological, or nuclear. If chemical or biological weapons are used, we must be ready to help with the diagnoses and follow-up care of these patients. Probably the greatest challenges to the radiology community will arise if the terrorist act involves a radiological or a nuclear explosive device. Understanding terrorists' goals of creating pandemonium and causing economic disruption is important. Radiology professionals need to be prepared to be resources for the medical community in providing patient care and for the community at large, especially if the terrorist attack involves detonation of a nuclear device, an attack on a nuclear power plant, or the use of a simple radiation dispersal device in a highly populated area. PMID- 17411521 TI - RADPEER quality assurance program: a multifacility study of interpretive disagreement rates. AB - PURPOSE: To develop and test a radiology peer review system that adds minimally to workload, is confidential, uniform across practices, and provides useful information to meet the mandate for "evaluation of performance in practice" that is forthcoming from the American Board of Medical Specialties as one of the four elements of maintenance of certification. METHOD: RADPEER has radiologists who review previous images as part of a new interpretation record their ratings of the previous interpretations on a 4-point scale. Reviewing radiologists' ratings of 3 and 4 (disagreements in nondifficult cases) are reviewed by a peer review committee in each practice to judge whether they are misinterpretations by the original radiologists. Final ratings are sent for central data entry and analysis. A pilot test of RADPEER was conducted in 2002. RESULTS: Fourteen facilities participated in the pilot test, submitting a total of 20,286 cases. Disagreements in difficult cases (ratings of 2) averaged 2.9% of all cases. Committee-validated misinterpretations in nondifficult cases averaged 0.8% of all cases. There were considerable differences by modality. There were substantial differences across facilities; few of these differences were explicable by mix of modalities, facility size or type, or being early or late in the pilot test. Of 31 radiologists who interpreted over 200 cases, 2 had misinterpretation rates significantly (P < .05) above what would be expected given their individual mix of modalities and the average misinterpretation rate for each modality in their practice. CONCLUSIONS: A substantial number of facilities participated in the pilot test, and all maintained their participation throughout the year. Data generated are useful for the peer review of individual radiologists and for showing differences by modality. RADPEER is now operational and is a good solution to the need for a peer review system with the desirable characteristics listed above. PMID- 17411522 TI - Coding and billing: the long road from ABC to W-2. PMID- 17411523 TI - What we most need to see. PMID- 17411524 TI - Selecting a new computed tomography scanner: things to consider. PMID- 17411525 TI - Practice management issues for radiology residents: focus on self-referral. PMID- 17411526 TI - Lobby day for radiologists in Virginia. PMID- 17411527 TI - Leaping frogs, P4PS, and P4P: not so strange bedfellows. PMID- 17411528 TI - It's only money... PMID- 17411529 TI - Resolved: that informed consent be obtained before screening CT. PMID- 17411530 TI - Anesthesia wars: lessons for radiology. PMID- 17411531 TI - The national institute of biomedical imaging and bioengineering and the extramural funding of imaging research. PMID- 17411532 TI - The risks of being sued and how much is paid. PMID- 17411533 TI - Practice guidelines: the radiology perspective. AB - Clinical practice guidelines have emerged as a reality for medical practitioners over the past 20 years. Although virtually all groups interested in the development of practice guidelines hope for improvements in patient care, secondary expectations vary widely among those using them. Their use in daily practice by physicians has met with resistance from barriers including concerns of "cookbook" medicine, a loss of autonomy, and increased professional liability. The recent experience of the ACR in addressing these challenges illustrates that physicians are receptive to steps perceived to mitigate the risks accompanying the use of guidelines as well as to efforts to increase their understanding of implementing guidelines in clinical practice. The experiences of other medical societies and an inventory of future trends reveal additional challenges associated with the use of practice guidelines, as third parties look to guidelines as points of reference for gauging the performance of health care providers. PMID- 17411534 TI - Commentary: ACR practice guidelines and technical standards. PMID- 17411535 TI - The manpower crisis in academic radiology: don't kill the milk cow for meat. AB - There is a manpower crisis in academic radiology departments. These departments cannot sustain their academic missions from clinical revenues alone. Salaries can' t be competitive with private practice, and the recruitment and retention of faculty members are compromised. The education of medical students, residents, and fellows and the clinical and basic research that sustains the specialty suffers. There is no simple remedy; academic departments need philanthropy from industry and private practice, more support from the government and the schools of medicine, and more efficient clinical practices. The future of our specialty is truly at stake. Academic departments are responsible for the great majority of training and technical innovation in the specialty. If academic departments cannot sustain their academic missions, the specialty of diagnostic radiology will certainly suffer. PMID- 17411536 TI - Resident duty hours: the rest of the story. AB - This article presents the complex set of considerations and influences that resulted in the Accreditation Council on Graduate Medical Education' s (ACGME) development of rules to govern the duty hours of residents. The process is demonstrative of the complexity associated with the self-regulation of graduate medical education. At the same time, the ACGME' s development of standards for resident duty hours suggests that self-regulation can be an effective alternative to governmental regulation. PMID- 17411537 TI - The business of radiology and the mission statement. AB - Practices with good management will outperform their peers and thrive in the future. This article is designed to expose and educate practicing radiologists to a powerful tool that can help design, create, and operate a business-oriented radiology practice. We introduce the mission statement as the most basic embodiment of business strategy. Mission statements and their derivatives are often at the heart of basic strategy formulations of successful organizations, yet they are rarely used effectively in radiology groups. There are differing approaches to the development of basic strategic statements, but careful and thoughtful attention to this formulation can provide worthwhile insight into an organization' s operations. Much of what is discussed is difficult or impossible to implement without the proper attitude and attributes in management. The help of trained outside professionals may even be required. Radiologists should embrace the concept of radiology as a business and move immediately to the next step: learning and applying modern business and management concepts to daily medical practice. PMID- 17411538 TI - Computed tomography and radiation: understanding the issues. AB - We are currently seeing increasing opportunities to improve patient care with computed tomography (CT). At the same time, we are challenged to use this technology wisely. In particular, we are being asked to balance the benefits against the risks, chiefly those of ionizing radiation. To do this, we must have a foundation from which to determine the relative risks. This foundation necessarily must be composed of several components. First, it is important to understand the patterns of use and increasing application of CT, particularly multidetector CT. In addition, it is helpful to be familiar with measures of radiation pertinent to CT and the doses provided by this modality. This foundation then provides a context in which to discuss the issue of low-dose radiation and cancer risk as well as potential changes in CT practice guidelines and regulation. It is with an understanding of these issues that radiologists and other radiology personnel can participate in an informed discussion with referring physicians and patients and continue to optimize the practice of CT. PMID- 17411539 TI - Diagnostic medical physicists and their clinical activities. AB - PURPOSE: The primary objective of this study was to obtain basic, descriptive information about medical physicists involved in diagnostic radiology-related activities, the diagnostic-related activities that they performed, and the time spent on these activities. METHODS: A survey was sent to a randomly selected sample of 1511 medical physicists from July through October 2001 using primarily e-mail methods; a total of 851 surveys was received, for a response rate of 56%. Of these, 427 were responses from physicists who do partly or only clinical diagnostic medical physics; it is this group for which results are presented. RESULTS: Fifty-four percent of the physicists who reported doing any clinical diagnostic medical physics performed clinical activities only in diagnostic medical physics. Fourteen percent of all those doing clinical diagnostic medical physics were women. Over 97% of the physicists doing clinical diagnostic medical physics reported having graduate degrees in physics; 53% had PhDs. The mean total weekly hours worked by physicists doing clinical diagnostic medical physics was 42. Medical physicists doing only clinical diagnostic activities reported working approximately 40 hours weekly, whereas those doing partly clinical diagnostic medical physics reported working 14 hours weekly in the field (approximately one third of their work time). Radiography and fluoroscopy, computed tomography, nuclear medicine, and mammography are all fields in which the majority of those doing any clinical diagnostic medical physics are active. Full-time physicists working only in diagnostic medical physics were responsible for a median of 25 units of equipment, compared with a median of 10 units for those working only partly in the field. Number of units evaluated, frequency of evaluation, and hours per evaluation were reported for almost 20 types of equipment. CONCLUSIONS: Medical physicists performing diagnostic clinical activities typically are responsible for a large number and wide variety of imaging equipment. It would be helpful to study their work further, focusing in particular on whether there is a shortage, as is true of diagnostic radiologists, and whether the variety of responsibilities creates strain. PMID- 17411540 TI - Quality and variability in diagnostic radiology. AB - Achieving and delivering optimal quality of care in radiology requires continual self-examination by the profession, particularly with regard to technical, interpretive, and communication skills. The importance of empirical data pertaining to quality and variability in radiology, the underlying causes of error, and the sources of variability are discussed. Key measures (e.g., receiver operating characteristics, kappa) and approaches (professional audits and peer reviews, surveys, inspections, and risk management programs) used in improvement efforts are reviewed, and data from key studies are highlighted. Diagnostic errors are important because of their connection to outcomes and the wide variability observed with modalities such as chest radiography and mammography. PMID- 17411541 TI - From errors to process improvement. PMID- 17411542 TI - Teacher or educational scholar? they aren't the same. AB - As a result of dramatic changes in the health care environment, the importance of the teaching mission to the future of radiology is gaining greater recognition. A new breed of "clinician educators" has a presence in most academic radiology departments. A clear understanding of the differences between teaching and educational scholarship is necessary for the purpose of defining the role of the clinician educator. For teaching to be considered scholarship, it must be public, susceptible to critical review, and accessible for exchange and use by other scholars; unless teaching has all three of these characteristics, it cannot be considered scholarship. Medical schools can improve the retention of their best teachers by defining tracks and criteria for rewarding teaching efforts. Universities, medical schools, academic medical centers, and professional organizations are vital components of the infrastructure needed to promote educational scholarship. They provide a community of education scholars who share, collaborate, and exchange work and promote the science of education. PMID- 17411543 TI - Reducing radiation to children: the resident's role. PMID- 17411544 TI - Purchasing computer systems need not be so difficult. PMID- 17411545 TI - Radiation safety. PMID- 17411546 TI - Being chair of a radiology department: observations from The Netherlands. PMID- 17411547 TI - What is a good radiologist? PMID- 17411548 TI - Diversity, pro and con. PMID- 17411549 TI - ACR 2004 gold medalists. PMID- 17411550 TI - Radiologist-patient contact during the performance of cross-sectional examinations. PMID- 17411551 TI - HIPAA and the law of unintended consequences. PMID- 17411552 TI - Venous access: deciphering the new codese. PMID- 17411553 TI - Turf wars in radiology: the overutilization of imaging resulting from self referral. AB - A recent report by the Medicare Payment Advisory Commission to Congress indicated that the utilization of diagnostic imaging is growing more rapidly than that of any other type of physician service. This has engendered concern among those who pay for health care. In this article, the authors review the role of self referral in driving up imaging utilization. A number of studies of the self referral factor in imaging have been conducted over the past three decades. These have consistently shown that when nonradiologist physicians operate their own imaging equipment and have the opportunity to self-refer, their utilization is substantially higher than among other physicians who refer their patients to radiologists. It has also been shown that the vast bulk of the recent increases in imaging utilization are attributable to nonradiologists who self-refer. The authors estimate that the cost to the American health care system of unnecessary imaging resulting from self-referral by nonradiologists is $16 billion per year. PMID- 17411554 TI - Should diversity be a factor in medical admissions? AB - The use of race, sex, and ethnicity in medical admissions represents one of the most important ethical and political issues currently before United States medical schools and residency programs. Critics of diversity-weighted admissions argue that academic merit is the only just basis for preferring one applicant over another, that underrepresented groups are difficult to define and constantly changing, and that using preferences to remedy past wrongs merely perpetuates discrimination. Proponents argue that past discrimination must be redressed, that the profession of medicine must produce physicians who mirror the population they serve, and that grades and test scores are not the only way of predicting who will be a good physician. When it comes to the role of diversity in medical school and residency admissions, there is no room for ignorance or apathy, because the futures of both medicine and radiology hang in the balance. PMID- 17411555 TI - The use of magnetic resonance imaging in breast cancer screening. AB - Screening mammography is a powerful tool for reducing breast cancer mortality. Mammography can often detect clinically occult, early-stage breast cancer that is amenable to successful treatment. However, mammography is not a perfect test and has lower sensitivity in young women and in those with dense breasts. Magnetic resonance imaging (MRI) has been shown to depict breast cancers that are occult to other forms of detection, including mammography. This has generated interest in the use of MRI for breast cancer screening. Although preliminary studies in highly selected populations show promise for the potential efficacy of breast cancer screening with MRI, there are many questions that should be addressed before this technique is offered to the general population. PMID- 17411556 TI - Retirement savings plans for radiologists: part 1--the options. AB - Employer-sponsored retirement plans are the primary savings vehicles used by radiologists to fund their retirements. A variety of retirement plans are available with guidelines, benefits, and restrictions specified by the Internal Revenue Code. In this article, we review and summarize the salient features of these plans. A second article, in an upcoming issue of JACR, explores the current status of radiologists' retirement plans. PMID- 17411557 TI - National radiology fellowship match program: success or failure? AB - In early 2003, with strong support from radiology leaders, the first national fellowship match was conducted in radiology. Most programs (358 programs in 9 categories) in the country participated, and 411 positions were filled out of 769 available in the match. Residents who selected to stay at their home departments were chosen before the match. A survey was conducted after the match to assess the satisfaction of chairpersons, program directors, and residents. Overall, the match was rated a success. However, a number of the pediatric fellowship programs remained outside of the match, possibly threatening its future. The initial radiology fellowship match that involved all fellowship specialties was deemed by residents and program leaders to be an overall success. Future matches will attempt to deal with such issues as incomplete participation. A significant problem is the continuing excess of offered fellowship positions over the number of applicants. PMID- 17411558 TI - A geologic survey of the medicare RBRVS system. AB - The Resource Based Relative Value System (RBRVS) was established over fifteen years ago in an attempt to bring order to the Medicare reimbursement system. The RBRVS relies on a complicated formula to determine how much each procedure is worth. This article uses an unusual approach to describe the components of the RBRVS equation, how those relative values are developed and managed, and how they relate to each other, even though each one is calculated differently. PMID- 17411559 TI - Lifelong learning and the maintenance of certification. AB - Lifelong learning is critical to radiology and the method by which, as professionals, we have kept up to date on new knowledge and developments. The American Board of Medical Specialists and the American Board of Radiology, along with other professional societies such as the Council of Medical Specialty Societies and the ACR, identified a need and an opportunity to serve the public and our profession through the Maintenance of Certification (MOC) initiative. These issues of lifelong learning, public concern, and MOC-and their interrelationship-are discussed. Lifelong learning is defined as a continuation of medical education with an ongoing process of professional development along with self-assessment, which enables physicians to maintain the requisite knowledge, skills, and professional standards. Continuing professional development can be grouped into three categories: formal, nonformal, and informal learning. Examples from the author's career illustrate key points. Future technologies and scientific discoveries affect radiology and make the commitment to lifelong learning even more critical. The challenge will be the maintenance of specialty certification and its components and competencies, along with the attainment of knowledge and the skills necessary in our unique practices for quality patient care in the evolving health care environment. These are important challenges to our boards, professional societies, specialty societies, institutions, and practicing community. It is critical to participate actively in these organizations and processes to continue development of the MOC initiative, because it is beneficial to patients, applicable to practicing physicians, and credible to public interests. PMID- 17411561 TI - The need for clinical trials in radiology. AB - The dependence of radiologic practice on habit and anecdotal evidence puts in doubt the appropriateness of care. Influences in the United States health-care system require that the specialty of radiology emphasize a transition to more appropriate care to reduce waste, thus improving patient outcomes and reducing cost. Understanding what constitutes the most appropriate care depends on improving the evidence basis for imaging practice. This can be accomplished through the participation of radiologists in multicenter clinical trials that are more pertinent, valid, reliable, and generalizable than past radiologic research. Improved training of residents and fellows in critical thinking, the conduct of research, and better appreciation of the value of research to the specialty, as well as increasing infrastructural support for imaging clinical trials, are necessary prerequisites for improving the appropriateness of radiologic practice. PMID- 17411562 TI - Disagreement in interpretation: a method for the development of benchmarks for quality assurance in imaging. AB - PURPOSE: To calculate disagreement rates by radiologist and modality to develop a benchmark for use in the quality assessment of imaging interpretation. METHODS: Data were obtained from double readings of 2% of daily cases performed for quality assurance (QA) between 1997 and 2001 by radiologists at a group practice in Dallas, Texas. Differences across radiologists in disagreement rates, with adjustments for case mix, were examined for statistical significance using simple comparisons of means and multivariate logistic regression. RESULTS: In 6703 cases read by 26 radiologists, the authors found an overall disagreement rate of 3.48%, with a disagreement rate of 3.03% for general radiology, 3.61% for diagnostic mammography, 5.79% for screening mammography, and 4.07% for ultrasound. Disagreement rates by radiologist for the 10 radiologists with at least 20 cases ranged from 2.04% to 6.90%. Multivariate analysis found that controlling for other factors, both differences among radiologists and across modalities, statistically significantly contributed to differences in disagreement rates. CONCLUSION: Disagreement rates varied by modality and by radiologist. Double reading studies such as these are a useful tool to rate quality of imaging interpretation and to establish benchmarks for QA. PMID- 17411563 TI - NHLBI and the NIH roadmap. PMID- 17411564 TI - The art of decision. PMID- 17411565 TI - Intensity modulated radiation therapy (IMRT)--a new approach to external beam RT. PMID- 17411566 TI - On getting the most out of residency. PMID- 17411568 TI - The nonsystem of American health care. PMID- 17411569 TI - ACR 2004 honorary fellows. PMID- 17411570 TI - Radiology and nuclear medicine: building a stronger partnership. PMID- 17411571 TI - NIH roadmap: course change for the big ship. PMID- 17411572 TI - Federal research report: the National Institute of Allergy and Infectious Diseases scientific and national security mission. PMID- 17411573 TI - Teleradiology: a legal perspective. PMID- 17411574 TI - Offshore teleradiology. AB - Radiologists are responsible for providing prompt emergency radiology interpretations 24 hours a day, every day of the year. As a result of the increasing use of multidetector computed tomography, emergency radiology has increased significantly in volume over the past 5 years. Simultaneously, radiologists are working harder during the day because of the workforce shortage. Although teleradiology services located in the continental United States have been providing efficient coverage until recently, they are now having increasing difficulty recruiting radiologists who are willing to work at night. Addressing this problem is "offshore teleradiology." With the increasing use of several enabling technologies--Digital Imaging and Communication in Medicine, the picture archiving and communication system, and the Internet-it is now possible to cover a domestic radiology practice at night from any location in the world where it is daytime. Setting up such a practice is nontrivial, however. The radiologists must all be American trained and certified by the American Board of Radiology. They must have medical licenses in every state and privileges at every hospital they cover. This article describes some of the details involved in setting up an offshore teleradiology practice. It also attempts to make a financial case for using such a practice, particularly in the current economic environment. PMID- 17411575 TI - The malpractice liability crisis, part 2: moving toward workable solutions. AB - As described in the previous article in this series, the United States is in the midst of a medical liability insurance crisis. Research conducted in the wake of similar previous insurance environments has shown that only a fraction of patient injury as a result of negligent medical care is addressed by today's tort malpractice system. This large reservoir of potential malpractice cases, coupled with the inevitable cycles of the economy and insurance markets, means that current action is mandatory to prevent another, even more serious crisis in the future. A variety of potential solutions have been proposed, including reform of the existing tort malpractice system, a no-fault compensation system, and so called hybrid systems that combine tort and no-fault. These mechanisms should be evaluated in terms of three key factors: (1) the adequacy of compensation to injured patients, (2) the prevention of future medical injury, and (3) their overall cost and administrative feasibility. Whatever reform mechanism or combination of mechanisms is pursued should represent an improvement on today's existing tort malpractice system. PMID- 17411576 TI - Retirement savings plans for radiologists, part 2: a comparison of academic and private practice retirement benefits. AB - A survey on retirement benefits was conducted involving 42 academic radiology departments and 42 private practice radiology groups. In this article, we present and discuss the survey results and provide recommendations for improving the retirement benefits for radiologists in both private practice and academic settings. A previous article, in last month's issue, provides details of how the various retirement plans mentioned in this article operate. PMID- 17411577 TI - Collaborating with hospitals: what radiologists need to know. AB - Less than a decade after fears about managed care drove medical students away from subspecialty training, the health care industry faces a deepening shortage of radiologists. Radiologists feel the shortage through busier days, longer hours, and recruitment difficulties. The shortage places many hospitals in a precarious position because diagnostic imaging is both a clinical necessity and a strong financial performer. However, radiology groups are prospering by redefining their relationship with hospitals and developing collaborative strategies. PMID- 17411578 TI - Breast imaging flow models. AB - Economically efficient and clinically effective breast imaging can be achieved through combinations and permutations of four fundamental breast imaging flow models: (1) the traditional flow model, (2) the online flow model, (3) the women's imaging flow model, and (4) the combination-care flow model. The structure, strengths, and weaknesses of these four individual breast imaging flow models are described and compared. PMID- 17411579 TI - The electronification of the radiation oncology treatment cycle: the promises and pitfalls of a digital department. AB - Radiation oncology departments are becoming more complex in terms of documentation, calculation, and delivery of therapy. Automation of such processes minimizes the likelihood of human error in each, and is clearly the direction in which the discipline is heading. The patient treatment cycle should be seamlessly integrated; unfortunately this is seldom the case given the different systems involved. We describe the Emory University experience, with cautions provided based on our lessons learned. PMID- 17411580 TI - Incorporating ACR Practice Guidelines, Technical Standards, and Appropriateness Criteria into resident education. AB - The ACR's Practice Guidelines, Technical Standards, and Appropriateness Criteria are useful tools in resident education. The Accreditation Council for Graduate Medical Education (ACGME) is increasing its focus on assessing educational outcomes in resident education to ensure that graduating residents have achieved competence to the level expected of new practitioners. The Appropriateness Criteria can be used to educate residents in cost-effective imaging practice, while the Practice Guidelines and Technical Standards are useful tools for educating residents in patient care. Incorporating these ACR resources into the resident curriculum and the goals and objectives of the training program provides a means of addressing the new ACGME requirements. PMID- 17411581 TI - The shortage of radiotherapy physicists. AB - The difficulties many radiotherapy facilities have in hiring new medical physicists indicate a shortage in the production and supply of these specialists. It appears that about 130 out of 350 new positions go unfilled each year. This projection matches well with the estimate that around 217 new medical physicists graduate per year. The problem is likely to become worse as a large cohort of practitioners reach retirement age in a couple of years. The radiotherapy physicists in the field tend to be working longer and harder, potentially leading to burnout or increases in errors. Addressing the difficulty in finding trained medical physicists, some facilities may start hiring untrained physicists or assigning technological personnel to perform the functions of a medical physicist, both of which could pose dangerous situations. While more physics undergraduates are showing interest in medical physics as a career, and medical physics graduate programs have increased enrollment by more than 30% over the last two years, further significant increases seem unlikely soon because of limited resources. Increasing resources (mostly faculty at the teaching universities) will require more research funding from governmental agencies--a situation unlikely in the near future. New mechanisms of funding medical post graduate education are needed. PMID- 17411582 TI - Residents' role in organized medicine. PMID- 17411583 TI - Is PACS worth the investment? it depends on your perspective. PMID- 17411584 TI - Medical physicists as radiation safety officers: a conflict of interest? PMID- 17411585 TI - The British National Health Service. PMID- 17411586 TI - Comment on "RADPEER quality assurance program: a multifacility study of interpretive disagreement rates". PMID- 17411589 TI - Isn't anybody paying attention? PMID- 17411590 TI - Reductionism and the failure of radiobiology. PMID- 17411591 TI - Medicare reform legislation: a new chapter in a long story. PMID- 17411592 TI - "We're from the government and are here to help you." Really! PMID- 17411593 TI - Compliance: not all fun and games. PMID- 17411594 TI - The new radiology workforce: changing expectations. AB - The zeitgeist of the new radiology workforce can best be described by a Bob Dylan song title: "The Times They Are A-Changin'." The new generation of physicians, although embracing the same foundations of medical practice as previous generations, places greater emphasis on personal satisfaction than its predecessors. Gone are the days when physicians operated as sole practitioners; today's workforce member is content to function in the role of "employee" in a trade-off for more lifestyle flexibility. This change has occurred not because of one specific factor but because of a change in the profession of medicine coupled with a combination of factors; familial responsibilities, avocational activities, and personal satisfaction have surfaced as motivating factors in choosing a profession. Today's workforce has a personal perception of success that may not be fulfilled solely by the contemporary practice of medicine. With the radiologist shortages that are now occurring and anticipated increased demand for staff radiologists, today's radiology workforce has helped shape the specialty into one that is altering its structure to attract and retain its workforce. PMID- 17411595 TI - Turf wars in radiology: other causes of overutilization and what can be done about it. AB - The overutilization of noninvasive diagnostic imaging (NDI) is a complex problem with many other aspects aside from self-referral. This article discusses (1) other causes of overutilization, (2) the rationales used by nonradiologist physicians to justify self-referring their patients for NDI, (3) the extent of self-referral by radiologists (called "autoreferral" by some), and (4) steps that could be taken to curb the overutilization of NDI in the United States. It is important for radiologists to be familiar with these issues and to support efforts to control overutilization. PMID- 17411596 TI - The Medicare Prescription Drug Improvement and Modernization Act of 2003: summary analysis and preliminary assessment of implications for radiology. AB - The Medicare Prescription Drug Improvement and Modernization Act of 2003 contains dozens of provisions of interest to radiology. Beyond enacting a new drug benefit, the bill contains hundreds of amendments affecting all aspects of Medicare payment policy. This article identifies those provisions most likely to affect radiological practice, and provides a preliminary assessment of the implications of these policies for radiologists. PMID- 17411597 TI - Residents' perceptions of MRI training in the United States. AB - PURPOSE: The primary objective of this exploratory study was to gain a better understanding of perceptions of magnetic resonance imaging (MRI) training by US radiology residents. METHODS: Two surveys were administered between September 2002 and June 2003 to nonprobability-based samples totaling approximately 2000 residents, under the auspices of the ACR's Resident Physician Section. The MRI Training Survey was administered in 2002, and it was supplemented by the Cardiac MRI Training Survey in 2003. The response rate for the MRI Training Survey was 15%, whereas the response rate for the Cardiac MRI Training Survey was 43%. RESULTS: The sample of residents surveyed comprised approximately 74% men; approximately 80% were in their third postgraduate year or higher. Eighty-five percent of residents reported the receipt of formal training in MRI physics. At least 60% did not have dedicated MRI rotations in either musculoskeletal or body MRI, and 89% lacked dedicated rotations in cardiac MRI. Seventy percent reported that they viewed overall MRI training as less than adequate; 94% stated that dedicated MRI training was warranted. CONCLUSIONS: This preliminary research suggests that current MRI training may be less than residents feel is needed in many training programs. New comprehensive, electronically disseminated core curricula, as well as a potential future mandate for minimum training requirements by the Accreditation Council for Graduate Medical Education's Radiology Residency Review Committee, may be necessary to strengthen MRI training at a time when the demand for radiology services is growing. PMID- 17411598 TI - Selected important characteristics for enlightened medical leaders. AB - An enlightened leader understands that unleashing the creativity and energy of the individuals in an organization is crucial to maximizing the potential of that group. A quality leader is more than a manager and invites the membership to help in the process of directing and caring for an organization and its people. A wise leader promotes communication, learning, innovation, and empowerment for all and always acts with integrity and professionalism. PMID- 17411599 TI - Promoting good work. AB - The workforce shortage confronting radiology renders it vital that radiologists explore the underlying features of good work, the kind of professional experience that makes people enjoy and feel proud of their performance. To thrive professionally, highly educated professionals such as radiologists need to feel that their work makes a difference, inspired by a clear sense of professional missions. PMID- 17411600 TI - The ACR imaging network: a retrospective on 5 years of conducting multicenter trials in radiology and plans for the future. AB - The ACR Imaging Network (ACRIN) provides a complete infrastructure to conduct multicenter clinical trials of medical imaging technologies as they relate to cancer. During its first 5 years of National Cancer Institute funding, ACRIN initiated work on 18 trials, a number of which are closed, in analysis, with results approaching publication. During its next funding cycle, ACRIN intends to pursue a more strategic approach to its selection of trials, focusing on key clinical questions in which imaging can improve the care of cancer patients. ACRIN represents a critical resource for the specialty in which all interested parties can participate. The results of ACRIN trials will improve the appropriateness of the use of imaging and benefit both those with cancer and those who are concerned that they might have the disease. PMID- 17411601 TI - After-hours coverage: problems and solutions. AB - Among the problems facing many radiology groups today is how to cover after-hours studies, because the demand is increasing while the number of available radiologists is still relatively low. There are a number of possible solutions, each of which has its own pros and cons, and no solution is right for every group. Recently, there have been a number of companies whose sole business is providing outside teleradiology coverage of after-hours radiology studies, sometimes referred to as "nighthawk" services. This article describes one group's decision-making process in choosing to hire a nighttime teleradiology provider as well as its subsequent experiences and ideas for future solutions. PMID- 17411602 TI - Selecting a new PET/CT system. PMID- 17411603 TI - Why radiology leaders fail. PMID- 17411604 TI - Research during radiology residency: challenges for the future. PMID- 17411605 TI - The Florida Radiological Society's efforts to correct the professional liability insurance crisis. PMID- 17411606 TI - Specialty or commodity? PMID- 17411608 TI - Meeting the professional liability crisis--doctors for medical liability reform. PMID- 17411609 TI - Working together, we can achieve our goals in 2004. PMID- 17411610 TI - Mammography and monkey bars. PMID- 17411611 TI - Tom Coburn, MD: a doctor in the house. PMID- 17411612 TI - Anatomy of malpractice defense, part 1: suit through discovery. PMID- 17411613 TI - ACR intersociety conference 2003: radiologist assistants and other radiologist extenders. AB - Improvements in imaging technologies are contributing to increases in the demand for radiology services. Efforts to match this rising demand are limited by the Centers for Medicare & Medicaid Services cap on resident positions and the challenges in obtaining appropriate visas and medical licenses for international medical graduates. This rising gap in the demand for and capacity to deliver radiology services has created a need for radiologist extenders. A variety of roles have been developed for these radiologist extenders, depending on the skills of the individuals and the subspecialty areas in which they work. Prominent among these are radiologist assistants, physician assistants, nurse practitioners, dosimetrists, and advanced-practice nuclear medicine technologists. Quality patient care is best accomplished when radiologist extenders function under the guidance of qualified radiologists, nuclear physicians, or radiation oncologists. PMID- 17411614 TI - ACR and ASRT development of the radiologist assistant: concept, roles, and responsibilities. AB - ACR and ASRT joined forces to address the workforce shortages of radiologists and radiological technologists with the development of a radiologist assistant. This paper discusses the rationale for the evolution and development of the radiologist assistant as designed by ACR and ASRT. With the profession of radiology experiencing workforce shortages among radiologists and radiological technologists many radiological groups were beginning to hire ancillary personnel to fill efficiency gaps in practices. Many academic institutions were in the process of considering programs to initiate advanced practice status for radiologic technologists. Several state legislatures were considering laws that would provide extended scope of practice technology with additional training. The ACR and ASRT entered into discussions to address these concerns in order to: (1) assure patient safety by allowing only properly trained personnel to provide radiological service to patients; (2) assure that these ancillary personnel are not allowed to practice without appropriate physician supervision; (3) assure that there are not 50 different state scope of practice laws based more on political expediency than quality patient care; (4) alleviate workforce shortages for radiological technologists by creating a professionally satisfying career path; (5) alleviate some of the time pressures placed on radiologists as a result of their workforce shortages. The ACR had a choice to lead on the development of the roles and responsibilities of the radiologist assistant or continue to allow the process to evolve as market and political pressures warranted. As a leader in the profession, the ACR believed that it was in the best interest of its members and the profession to be at the table with the development of the roles and responsibilities of the radiologist assistant to assure the best for our patients. The roles and responsibilities of the RA as approved by the ACR Council are presented. PMID- 17411615 TI - Radiologist assistant certification. AB - The American Registry of Radiologic Technologists (ARRT) is the largest national, voluntary certification organization for radiologic technologists, with 235,000 registered technologists and an annual examination volume of 20,000 to 25,000. The ARRT's board is appointed by the ACR (four members) and the American Society of Radiologic Technologists (five members). The ARRT is in the process of developing a certification for radiologist assistants. Standards of education, ethics, and examination will be adopted to identify individuals qualified to assume this new role. The program is scheduled to be available in the fall of 2005. PMID- 17411616 TI - The likely effects of radiologist extenders on radiology training. AB - Manpower shortages have led many radiologists to consider using radiologist extenders, and the American Society of Radiological Technologists and the ACR have responded by formulating the radiology assistant (RA) program. There is no doubt that the RA program is becoming a reality or that it can be a boon to radiologists and physician radiologist trainees in freeing up faculty members' time, freeing trainees from repetitive tasks, and providing limited instruction in RAs' areas of expertise. Patients will likely accept RAs and trainee physicians equally. A financial analysis of using residents and physician extenders in anesthesia suggests that residents may be a cheaper labor source in those institutions not at the cap for resident positions. PMID- 17411617 TI - Reimbursement risks with radiologist extenders: there is no free lunch! AB - There is great allure in the concept of using qualified health care providers to assist radiologists and radiation oncologists, increasing efficiency and possibly even improving patient care delivery. However, physician services are most commonly reimbursed under a system that is resource based, and the physician work and practice expense components of reimbursement for existing procedure codes are periodically reexamined to ensure their appropriate rank in this "relative value system." Also, as new codes are developed, demonstrable physician work and practice expenses will determine the relative values for the new procedures. In both cases, the type of individual who actually performs different portions of a procedure will determine the reimbursement level. In addition, the total reimbursement must be appropriately apportioned between the physician involved and the facility where the service is delivered. This article examines some of the potential impacts on procedure coding and radiologist and radiation oncologist reimbursement schedules if physician extenders perform work previously performed by physicians. It also examines possible shifts in reimbursement from physician to facility if an extender is employed by a facility. PMID- 17411618 TI - Physician assistants in academic radiology: the Harborview experience. AB - We describe a model of how physician assistants can be used in an academic medical center to expand radiologist productivity, and to enhance the departmental academic and educational missions. At Harborview Medical Center, following a training program and graduated responsibility under supervision, physician assistants provide initial interpretation of radiology studies, consultation to referring physicians, and perform less complicated interventional procedures. Acceptance of physician assistants by the radiologists, radiology residents, and referring physicians has been high. Although the impact of physician assistants on departmental clinical productivity is difficult to measure, our data suggest that radiologists are more efficient when physician assistants are assigned to service, both in terms of numbers of studies interpreted, and timeliness of reporting and billing. As a result of the success of our program, we believe that physician assistants can have an important role in radiology practice. PMID- 17411619 TI - Cost-effectiveness model for first-trimester versus second-trimester ultrasound screening for Down syndrome. AB - PURPOSE: To develop a cost-effectiveness analysis model from the perspective of Medicare reimbursement to evaluate the costs and potential risks involved in performing second-trimester genetic sonography following the first-trimester sonographic measurement of nuchal translucency and serology for Down syndrome screening. METHODS: Three clinical screening algorithms were constructed that detailed the diagnostic evaluation of the target population by using first trimester or second-trimester ultrasound and appropriate serologies or first trimester and second-trimester screening in combination. The cost analysis was then created by using a computer spreadsheet program by applying Medicare reimbursement, the prevalence of Down syndrome, and reported sensitivities of first-trimester and second-trimester ultrasound and analytes for Down syndrome for each clinical algorithm. Medicare Current Procedural Terminology codes, total relative value units, and payments for first-trimester and second-trimester ultrasound, chorionic villous sampling, amniocentesis, and serum analytes were obtained from the Medicare Part B Washington 2002 Provider Disclosure Report. RESULTS: At any given prevalence of Down syndrome, first-trimester screening is always slightly less expensive to society than the other two models for both total cost and cost to diagnose each case of Down syndrome. Even if second trimester screening were 100% sensitive, the sensitivity of first-trimester screening would have to fall below 55% for model 2 to be cheaper than model 1. Combining both first-trimester and second-trimester screening was substantially more expensive than models 1 or 2. More iatrogenic fetal deaths occur with combined screening than with either first or second trimester screening alone. CONCLUSIONS: Screening using first-trimester ultrasound and serologic markers to screen for Down syndrome is always slightly less expensive to society than second trimester serologic and ultrasound screening. However, there is a significantly increased risk for iatrogenic fetal death if second-trimester genetic sonography is performed following normal first-trimester screening using currently accepted risk ratios. Patients should be counseled appropriately with this information, because an individual's circumstances will affect that person's perception of risk and subsequently affect his or her decision making. PMID- 17411620 TI - Effects of the 2003 match on the radiology fellowship application process. AB - PURPOSE: To explore the effects of the newly instituted 2003 radiology fellowship match on the fellowship application process. METHODS: Survey questionnaires were distributed to third year radiology residents during the March 2003 AFIP session. One hundred fourteen surveys were returned and reviewed. The questionnaire included whether the resident applied to fellowship programs and why, to which subspecialty they applied and whether the match had complicated or facilitated their application process. All questions (except for the subspecialty selected) were multiple choice and residents were encouraged to provide additional comments. Some residents did not answer all the questions. RESULTS: Seventy-five percent of residents applied to fellowship programs and 25% did not. Of those who did apply, the match complicated the application process for 58%, facilitated the process for 7%, did not effect 34% and both complicated and facilitated the process for 1%. Twenty-eight percent of residents for whom the match had no effect accepted fellowship positions outside the match. Complications included lack of uniform program participation in the match, absence of uniform acceptance of a universal application, lack of program awareness regarding the match timeline and structure, and increased monetary expense and time commitment involved with the match. The internal filling of fellowship positions was valued by some residents and viewed as unfair by others. The three most common fellowships applied to were musculoskeletal imaging, neuroradiology and body imaging. CONCLUSIONS: Overall, residents considered the match as unfavorable. Complications relating to the newness of the match could be resolved, inherent problems must be accepted, while other issues need further evaluation. PMID- 17411621 TI - Patient radiation ID cards. PMID- 17411622 TI - Leading your organization through a successful software implementation has little to do with the technology. PMID- 17411623 TI - Insight into the fiscal year 2005 and technological initiatives at the National Cancer Institute. PMID- 17411624 TI - Physician and resident unions: are we headed in the right direction? PMID- 17411625 TI - The system does work. PMID- 17411626 TI - William G. Jones, MD. PMID- 17411627 TI - ED Weber, DO. PMID- 17411628 TI - Self-referral: bad health care at any cost. PMID- 17411629 TI - The continuing diffusion of magnetic resonance technologies. PMID- 17411630 TI - Clinical IR: it's now or never. PMID- 17411631 TI - The other shoe: Medicaid. PMID- 17411632 TI - Fifth National Forum on Biomedical Imaging in Oncology. PMID- 17411633 TI - CT colonography and virtual reimbursement. PMID- 17411634 TI - Contracts in radiology practices: contract types and key provisions. AB - A contract between a radiology group and its physician member(s) provides the foundation for the professional relationships in a group practice. The parties are not in positions of parity; contract provisions are structured to maintain the primacy of the group over the individual members. An integration clause should be included to preclude reliance on communications that are not memorialized by the language of the contract. Precise, unambiguous terms must be used to convey the intentions of the parties. The contract should have a clear date of initiation and, if applicable, an effective date of termination. Mechanisms for termination and modification should be expressed clearly to minimize the risk of judicial interference. The method of determining and adjusting the salary and other benefits of the radiologist should be stated, as consideration is necessary to support the existence of a legal contract. The obligations of the radiologist to the practice are often stated in general terms to maximize the group's flexibility. Finally, other key clauses that are discussed in the paper should be incorporated into the contract. PMID- 17411635 TI - Exclusive hospital-based service agreements: what radiologists need to know. AB - This article provides radiologists with the information that they need to know to participate meaningfully in negotiating or renegotiating an exclusive hospital based radiology service agreement. It discusses the contract negotiation process, including how to identify and prioritize contract objectives, and how to assess and create bargaining leverage. Options for achieving contract longevity, for resolving "turf" issues and for achieving financial objectives are also addressed. The article further explains the key regulatory issues that shape exclusive hospital-based radiology service agreements, including antitrust, fraud and abuse, Stark Law, HIPAA, tax, and Medicare reimbursement considerations. The author discusses the contract negotiation process from both the radiology group and hospital perspectives. He suggests that successful negotiation will depend on "fitting" the group's contracting agenda with the hospital's priorities, organizational structure, culture and resources. PMID- 17411636 TI - Relationship between HMO market share and the diffusion and use of advanced MRI technologies. AB - BACKGROUND: Financial incentives associated with managed care may shift incentives associated with the adoption of new medical technologies. This study examined whether managed-care activity was associated with the adoption rate of magnetic resonance imaging (MRI) equipment during the 1990s. DATA AND METHODS: Data from three nationwide "censuses" of MRI sites conducted in 1993, 1997, and 1999 were used. The number of MRI sites and magnets; magnet field strength; MRI procedures; the use of contrast media; and the presence of power injectors, echoplanar imaging, cardiac MRI, and interventional MRI were measured in each of 322 metropolitan statistical areas each year. Regression analysis was used to assess the relationship between area MRI availability and overall area health maintenance organization (HMO) market share, controlling for potential confounders. RESULTS: Areas with higher HMO activity had markedly lower adoption and use of MRI. By 1999, high-HMO areas had about 40% fewer MRI scanners per 100,000 people than low-HMO areas (1.02 vs. 1.73, P < .01). High-HMO areas had fewer 1.5-T scanners than low areas in all 3 years and tended to use contrast media less often in 1993 and 1997 (all P < .01). There were statistically insignificant trends toward less availability of echoplanar imaging, cardiac MRI, and interventional MRI in high-HMO areas. CONCLUSIONS: The fact that managed care is associated with the slower adoption of MRI and less availability of some of the most advanced MRI equipment suggests the need for attention to the potential for managed care to have important effects on the quality of care and health care spending by influencing technology growth. PMID- 17411637 TI - Subspecialty training and certification for radiation oncology. AB - To address the recurring issues regarding subspecialty training and certification in radiation oncology, and using pediatric radiation oncology as an example, the authors considered the problem of inadequate case material for resident teaching. Potential solutions to the identified problems are addressed, and the roles of oversight committees and regulatory bodies are clarified. The problem of the nonuniform distribution of pediatric case material across residency programs cannot be solved by removing pediatric radiation oncology from the core radiation oncology educational curriculum or by offering fellowship training to those who may select it. As a result, the authors believe that subspecialty training and certification in radiation oncology is not a near-term possibility. PMID- 17411638 TI - Motivation and compensation in academic radiology. AB - As radiologists are increasingly faced with the challenges of rising demand for imaging services and staff shortages, the implementation of incentive plans in radiology is gaining importance. A key factor to be considered while developing an incentive plan is the strategic goal of the department. In academic radiology, management should decide whether it will reward research and teaching productivity in addition to clinical productivity. Various models have been suggested for incentive plans based on (1) clinical productivity, (2) multifactor productivity, (3) individual productivity, (4) section productivity, and (5) chair's discretion. Although fiscal rewards are most common, managers should consider other incentives, such as research time, resources for research, vacation time, and recognition awards, because academic radiologists may be motivated by factors other than financial gains. PMID- 17411639 TI - Improving the quality of radiology reporting: a physician survey to define the target. AB - PURPOSE: Our long-term goal is to improve the quality of reports in radiology imaging interpretation. The rationale for this project focused on identifying the characteristics of a high-quality report from the perspective of referring physicians and radiologists. METHODS: We undertook a survey of physician faculty at a large Midwestern academic medical center (including university, children's, veteran's, county and private practice hospitals) regarding radiology report quality concepts. RESULTS: Using a 5-point Likert scale, >95% of respondents indicated the highest importance rating (score=5) for radiology report characteristic "Accurate," with mean score of 4.94. Seventy-eight to 83% of respondents considered "Clear," "Complete" and "Timely" to have the highest importance rating, with means of these scores between 4.73 and 4.79. Somewhat less desirable characteristics included "Well-organized" and "Mentions pertinent negatives"-though radiologists tended to think the latter was less important than did all other categories of physician respondents. The single greatest problem area in reporting is lack of timeliness. Using a 10-point Likert scale, respondents gave a median score of seven for overall satisfaction with current reporting. CONCLUSIONS: For high-quality radiology reporting, accuracy is most important. Clarity, completeness and timeliness are also very important. Radiologists tend to consider mentioning pertinent negatives as less important than do referring physicians; otherwise, respondents from different specialties largely agreed on which characteristics are most important for high-quality reports. There is room for improvement in physician satisfaction with radiology reporting. PMID- 17411640 TI - Turf wars in radiology: the quality of interpretations of imaging studies by nonradiologist physicians--a patient safety issue? AB - Previous publications have demonstrated that in side-by-side testing, radiologists clearly outperform nonradiologist physicians at interpreting plain radiographs. Other articles, mostly in the nonradiologic literature, have shown that in actual practice settings, nonradiologist physicians make frequent errors in image interpretation. On the other hand, several other studies have concluded that such errors are infrequent and generally do not affect patient outcomes. However, most of the latter studies have had methodologic flaws. Radiologists need to be aware of the evidence that exists in the medical literature on errors made by nonradiologists in image interpretation and to bring it to the attention of hospital authorities or medical policy-making groups that are charged with improving patient safety. PMID- 17411641 TI - How do training, education, and experience affect quality in radiology? AB - The quality of care is attracting increasing attention from payers, regulators, and consumers. The assumption that training, education, and experience are major determinants of quality and safety permeates the health care delivery system. However, the relationship between quality and training, education, and experience is neither straightforward nor well documented, particularly for the practice of radiology. A recent Institute of Medicine report questioned the effectiveness of the existing system for the education and training of health care professionals. In this article, "quality" in radiology is defined, and evidence for a relationship between quality care and experience and traditional continuing medical education is reviewed. Alternative approaches to education and training, with an emphasis on systems rather than individuals, are elucidated. The nature of radiologist training, education, and experience should be reexamined and adjusted to meet the changing demands of the health care delivery system. PMID- 17411642 TI - Resuscitating the lecture. PMID- 17411643 TI - Radiology training in Europe. PMID- 17411644 TI - Magnetic resonance imaging at 3 Tesla: time to begin, again. PMID- 17411645 TI - Timing of the oral radiology boards. PMID- 17411646 TI - Information technology vendors: unwitting facilitators of medical errors? PMID- 17411647 TI - Tort reform and you: winners and losers, friends, foes, and facts of life. PMID- 17411648 TI - Healthy academic radiology departments ensure the future of the specialty. PMID- 17411649 TI - "Hood ornament" or "V-10 engine"? Myths and realities regarding physician scientists in academic radiation oncology departments. PMID- 17411650 TI - Abdominal aortic aneurysms and the Pulitzer Prize. PMID- 17411651 TI - Anatomy of malpractice defense, part 2: trial and beyond. PMID- 17411652 TI - Comparative increases in utilization rates of ultrasound examinations among radiologists, cardiologists, and other physicians from 1993 to 2001. AB - PURPOSE: To ascertain changes in the utilization rates of diagnostic ultrasound among radiologists, cardiologists, and other physicians in recent years. METHODS AND MATERIALS: The nationwide Medicare Part B databases for 1993 and 2001 were searched in all ultrasound Current Procedural Terminology 4 codes, except for ophthalmic ultrasound and supervision and interpretation codes. Ultrasound examinations were categorized as general, vascular, breast, obstetric (very low in the Medicare population), and echocardiography. Using the Medicare physician specialty codes, utilization rates per thousand Medicare beneficiaries per year were calculated for radiologists, cardiologists, and other physicians for all codes in the five aforementioned categories. RESULTS: Radiologists performed 24% of all ultrasound examinations in 2001. The overall utilization rate of ultrasound examinations among radiologists increased from 132.9 per thousand Medicare beneficiaries in 1993 to 166.3 in 2001, a 25% increase. Among cardiologists, the rate increased from 190.3 in 1993 to 356.1 in 2001, an 87% increase. The vast bulk of these examinations were echocardiograms, but cardiologists also had some involvement in vascular ultrasound. Among other physicians, the rate increased from 116.9 in 1993 to 167.0 in 2001, a 43% increase. The areas of greatest involvement by these other physicians were echocardiography, vascular ultrasound, and general ultrasound. Of the five ultrasound categories, echocardiography had by far the highest volume. CONCLUSION: Nonradiologist physicians utilize ultrasound at much higher rates than radiologists, primarily reflecting the influence of echocardiography. Between 1993 and 2001, the ultrasound utilization rate grew over 3 times as rapidly among cardiologists as among radiologists; the rate among other physicians grew almost twice as rapidly as it did among radiologists. This raises the concern that self-referral may be leading to higher utilization and costs. PMID- 17411653 TI - Contracts in radiology practices: breaches and remedies. AB - Contracts between radiology groups and their physician members are often ambiguous. Key clauses may not be precise as to the intent of the contracting parties. For example, the requirements for a group member to achieve shareholder status may be discussed but not reduced to a written form. Other contract provisions, such as termination or noncompete clauses, may be subject to different interpretations. The ambiguities of these provisions often generate disparate expectations regarding the parties' obligations to one another. When this occurs, the results may vary from disappointment to litigation. This paper discusses the causes and consequences of common breaches of radiology contracts. The types of remedies that may be available to the parties of the contract are also enumerated, and case law is cited to illustrate the challenges that radiology groups and their members may encounter. Finally, alternative forms of dispute resolution are discussed. PMID- 17411654 TI - Essential practice performance measurement. AB - The objective of this paper is to provide an overview of how to develop and implement a performance measurement system in radiology departments. Although an extensive literature review (PubMed, MEDLINE, etc) was carried out to search for relevant published scientific papers, the number of publications regarding performance indicators in radiology departments was very limited. The present paper reflects the current approach to performance measurement in health care services based on the available literature, which may be applied to the field of radiology. Performance indicators are tools that evaluate an organizations progress toward its goals . In radiology, in addition to finance, other aspects that affect the functioning of the organization, such as clinical productivity and patient satisfaction, also need to be assessed. The main categories of indicators adopted in radiology departments include: (1) productivity, (2) finance, (3) patient safety, (4) access, and (5) customer satisfaction. Once specific indicators in each of these categories are selected, the data collection methods should be incorporated into the routine departmental processes. Information obtained should be made available to all stakeholders via various media. In conclusion, performance indicators establish a common denominator in order to make comaparisons of the organization's performance over time. To improve the quality of services, these indicators should be benchmarked, i.e., the processes should be compared to the best in the field. PMID- 17411655 TI - The business of radiology: cost accounting. AB - Radiology practices confront questions of resource allocation every day. Unfortunately, practices frequently fail to adequately analyze revenues and expenses, which are at the heart of success or failure in any business endeavor. Cost allocation problems permeate nearly all aspects of cost analysis and accumulation and exist throughout all types of private-sector and public-sector organizations. "Managerial" or "cost" accounting is the discipline concerned with measuring and assigning the costs of delivering services or producing products. In contrast to financial accounting, management accounting produces relevant information for internal decision making and in general is designed to answer a firm's specific operational questions. Because costs play such a critical role in deriving and planning for revenues and profits, managerial accounting is in large part devoted to measuring and accumulating costs with the aims of control and continuous cost reduction. Because radiologists' salaries are at record highs, when accounting for a practice's clinical activities, such as the provision of mammography services, some allocation of radiologist costs themselves must be made, or the practice will not be able to achieve its goal of efficient allocation of resources. Whatever cost-accounting method is used should be specific enough to allow the differentiation of costs to as detailed a level as necessary for the strategic decision at hand. It is imperative that a practice use some rational method to gather and analyze costs and that management then use these data in decision making. Successful practices will be those most aware of their costs and the minimum acceptable reimbursements necessary for their success. PMID- 17411656 TI - Changes in graduate medical education: a view from radiation oncology. AB - The accreditation process of graduate medical education is changing. This change presents new challenges to teachers of radiation oncology. With the help of the ACR and other institutions, radiation oncology is well positioned to meet this challenge. PMID- 17411657 TI - Why don't female medical students choose diagnostic radiology? A review of the current literature. AB - While the number of women entering medical schools is approaching 50% nationally, women continue to be underrepresented in a number of specialties including diagnostic radiology. While diagnostic radiology has many characteristics that might be desirable to women, such as reasonable call hours, flexible scheduling, and high salaries, women still do not choose diagnostic radiology as a career. This article examines the literature to discern possible reasons for why women are entering diagnostic radiology at a lower rate. We address trends among women in academic medicine, which resemble trends among women in diagnostic radiology, and examine the effects of gender and socialization in medical school on specialty choices among women. The current literature suggests a constellation of factors may be responsible for the gender differences in diagnostic radiology. We suggest that further research is needed to elucidate why women do not seem to be choosing diagnostic radiology as frequently as one might predict based on the lifestyle of diagnostic radiologists and the numbers of women currently entering medical school. Once these reasons are made clear, it will be possible for residency program directors and medical schools to ensure that women are making informed specialty choices, whatever those choices may be. PMID- 17411658 TI - Enhancing research in academic radiology departments: recommendations of the 2003 Consensus Conference. AB - Opportunities for funded radiologic research are greater than ever, and the amount of federal funding coming to academic radiology departments is increasing. Even so, many medical school-based radiology departments have little or no research funding. Accordingly, a consensus panel was convened to discuss ways to enhance research productivity and broaden the base of research strength in as many academic radiology departments as possible. The consensus panel included radiologists who have leadership roles in some of the most well-funded research departments, radiologists who direct other funded research programs, and radiologists with related expertise. The goals of the consensus panel were to identify the attributes associated with successful research programs and to develop an action plan for radiology research on the basis of these characteristics. PMID- 17411659 TI - The added value of screening mammography in improved screening for other cancers. AB - As part of day-to-day practice, the most frequent role of the radiologist is to provide diagnostic information derived from imaging that will help the clinician better manage patient care. In essence, radiologists are reimbursed for the information we provide. The value of diagnostic information, however, extends beyond strict clinical and monetary value. The objectives of this paper are to provide the framework for understanding the "value" of diagnostic imaging. The article focuses on the application of these concepts using examples from the literature with particular emphasis on screening and suggests using a screening test experience as a means of improving health behavior. PMID- 17411660 TI - Why study physics? PMID- 17411661 TI - Acquiring a radiology information system for your practice: current state and future trends. PMID- 17411662 TI - Are there shortcuts in our search for the holy grail? PMID- 17411663 TI - Jack of all trades, master of none: why do a fellowship? PMID- 17411664 TI - Comment on article by Levin and Rao. PMID- 17411665 TI - Economists, weather forecasters, storms, and cliffs. PMID- 17411667 TI - Emerging technologies in radiation oncology and nuclear medicine: an overview of their use and regulation. PMID- 17411668 TI - Multidisciplinary care and emerging technologies: optimizing patient care or truce in a turf battle? PMID- 17411669 TI - Money, message, and political advocacy. PMID- 17411670 TI - Oh no! Not another acronym? PMID- 17411671 TI - Complaints. PMID- 17411672 TI - Clinical operations management in radiology. AB - Providing radiology services is a complex and technically demanding enterprise in which the application of operations management (OM) tools can play a substantial role in process management and improvement. This paper considers the benefits of an OM process in a radiology setting. Available techniques and concepts of OM are addressed, along with gains and benefits that can be derived from these processes. A reference framework for the radiology processes is described, distinguishing two phases in the initial assessment of a unit: the diagnostic phase and the redesign phase. PMID- 17411673 TI - A historical perspective of the radiation oncology workforce and ongoing initiatives to affect recruitment and retention. AB - The field of radiation oncology has become exceptionally technologically driven and multidisciplinary over the past two decades. This specialty of medicine is one that requires not only the command of highly complex modalities but also the assembly of a competent and expertly skilled team of medical professionals. Although the profession has grown tremendously in past years, the workforce has not been able to meet the demands of the practice. A significant shortage of radiation therapists, dosimetrists, and oncology nurses exists in the United States today and will almost certainly increase in severity over the next several years. A similar crisis has been seen in several other countries, most notably Canada and Australia, and has contributed to prolonged delays in cancer treatment for many patients. PMID- 17411674 TI - Turf wars in radiology: the quality of imaging facilities operated by nonradiologist physicians and of the images they produce. AB - There is considerable emphasis currently placed on the need to reduce medical errors and improve patients' safety. Because diagnostic imaging is such an important part of the workup of most medical conditions, one way to help achieve this goal is to insist that imaging facilities and the imaging examinations they produce meet high standards of quality. Published studies have shown that radiology facilities and radiologists consistently generate higher quality images than most other medical specialties. Radiologists should bring this evidence to the attention of payers and medical policy makers in their areas who are responsible for decisions affecting patients' safety. PMID- 17411675 TI - Professional resource cost of body CT examinations: analysis of interpretation costs in different patient populations. AB - PURPOSE: To study professional cost and cost drivers in body computed tomography (CT), and to compare the professional cost associated with similar CT studies performed on patients with dissimilar diseases. METHODS: A time and motion study was undertaken to measure the allocation of physician time in body CT. The hourly cost of radiologists' clinical practice was calculated. The physician cost associated with individual CT patients was then calculated according to the amount of time radiologists devoted to each patient. Technical costs and hospital overhead were not included. RESULTS: The hourly cost of the clinical practice of attending radiologists, cross-sectional imaging fellows, and senior radiology residents was $293, $35, and $35 respectively. The average professional cost for CT examination of an abdominal pain patient with abdominal and pelvic imaging was $63. The average professional cost for CT examination of an oncology patient with abdominal and pelvic imaging was $90. The cost per relative value unit (RVU) was $25 for abdominal pain patients and $35 for oncology patients. CONCLUSION: Similar CT studies performed on patients with dissimilar diseases result in differing professional costs. PMID- 17411676 TI - The Canadian health care system: past, present, and future. AB - This review offers American radiologists the information required to understand the Canadian health care system. It includes a brief history of the Canadian system and describes the relationship between health care and the Canadian constitution. A comprehensive appraisal of current conditions addresses the performance of the Canadian system and provides an overview of policy and planning responsibilities for health care across Canada. It includes not only the roles and responsibilities of the federal government and of the provincial and territorial governments in health care, but also those of professional organizations. In this context, significant differences between Canada and the United States are discussed, including: the development of professional fees; billing mechanisms and audits; and medicolegal and turf issues, such as self referral and imaging by nonradiologists. A description of professional organizations includes the central responsibility of the Royal College of Physicians and Surgeons of Canada for professional training and development, and of national and provincial medical organizations. There are descriptions of medical schools and research organizations as well as federal organizations responsible for health information and technology assessment. Problems encountered in recent years because of the curtailment of health care spending are addressed. The future of Canadian health care is considered in light of a recent report titled Building on Values: The Future of Health Care in Canada. Issues of particular interest to radiologists are emphasized. PMID- 17411677 TI - Changing radiology resident education to meet today's and tomorrow's needs. AB - Radiology education has evolved significantly in the past several decades, but there is considerable room for improvement. While some improvements depend on the availability of new technology, others are low tech, requiring merely that we be aware of and apply findings in the education literature. We teachers of radiology could benefit from recent studies investigating the processes underlying adult learning to improve the methods we use to train residents and fellows, and to improve the way we teach our colleagues and even ourselves. In this review, adult learning and the expert learner are briefly discussed, as are the important elements of effective learning tools. The strengths and weaknesses of radiology education as it is commonly practiced today are explored. Specific suggestions for improving education in radiology using both low tech and high tech methods are then discussed. PMID- 17411678 TI - The polity of academic medicine: status of faculty governance. AB - How should academic medical departments be governed? Models of governance span a wide spectrum between autocracies, in which important decisions are made by a single individual, to democracies, in which each member enjoys a voice in decision making. Despite the fact that more participatory governance models are the norm in practice settings outside of academia, many academic physicians seem to take an autocratic model more or less for granted. In fact, however, most medical schools and universities do not mandate a governance system that vests authority in a powerful chairperson, and departments frequently enjoy more latitude than they suppose in determining how to govern themselves. Because an organization's effectiveness is powerfully influenced by its governance structure, academic physicians should give serious consideration to this subject, to ensure that academic medicine is well prepared to meet the many challenges now before it. PMID- 17411679 TI - Residency training data: what do the numbers tell us? AB - The Accreditation Council for Graduate Medical Education (ACGME) regularly collects data on accredited residency programs that are useful for purposes of comparing resident demographics, turnover rates of program directors, and trends in program numbers and accreditation status. These data show that there are relatively fewer women in radiology residencies compared with other nonsurgical residencies. Women make up only 25% of core radiology residents. Further investigation is warranted to determine why this is so, because it will likely affect the ability to recruit female radiologists into leadership positions. Considering ACGME-accredited programs, those radiology subspecialties with the highest percentages of female trainees are pediatric radiology (45%) and nuclear radiology (44%). The turnover rate of radiology program directors is high (19%) relative to other specialties (14% on average), and this is being addressed in part by a change in the radiology program requirements. Other factors associated with this high turnover rate may also need to be addressed. PMID- 17411680 TI - Pelvic sonography: OB or not OB. PMID- 17411681 TI - State-of-the-art ultrasound imaging technology. PMID- 17411682 TI - Electronic point-and-click radiation oncology weekly examination notes. PMID- 17411683 TI - ACR chapter resident sections: getting involved locally. PMID- 17411684 TI - Re: "Improving the quality of radiology reporting: a physician survey to define the target" (J Am Coll Radiol 2004;1:497-505). PMID- 17411685 TI - Both sides of the aisle. PMID- 17411686 TI - Whither cardiovascular and interventional radiology? PMID- 17411687 TI - Unnecessary imaging and radiation risk: the perfect storm for radiologists. PMID- 17411688 TI - A primer on e-mail etiquette. PMID- 17411689 TI - Evidence-based medicine: ready for prime time? PMID- 17411690 TI - A large vision with a small focus: from the organ to the cell. PMID- 17411691 TI - Clinical practice of interventional and cardiovascular radiology: current status, guidelines for resource allocation, future directions. AB - The practices of interventional radiology and interventional neuroradiology are centered on high-quality direct patient care. These subspecialties have long histories of innovative care that has often revolutionized the treatment of disease and illness. More recently, however, this success has brought about competition from former referring physicians as they have gained access to technology and training that will enable them to obtain credentials for procedures that were formerly in the exclusive domain of interventionalists. Unfortunately, many interventional radiologists find themselves ill-equipped to compete for referrals. This is primarily because many interventional radiology practices lack complete clinical practices, which are critically important in facilitating referrals from the nonspecialists. Accordingly, this document details the critical importance of a complete clinical practice and further outlines the steps required to achieve this goal. PMID- 17411692 TI - HIPAA compliance: the law, reality, and recommendations. AB - The physicians of today and tomorrow face the most daunting set of regulations ever imposed on the practice of medicine. Through the passage of the Health Insurance Portability and Accountability Act of 1996 (HIPAA), the federal government has thrust its regulatory authority into three of the most controversial and cutting-edge issues in medical practice management: privacy, electronic transactions, and security. Through practical insights, the authors' goal is to introduce physicians to HIPAA's basic tenets, the evolutionary nature of the regulations, and the concept that they can manage HIPAA without bankrupting their practices or sealing themselves away from their patients. PMID- 17411693 TI - CPT coding by interventional radiologists: a multi-institutional evaluation of accuracy and its economic implications. AB - PURPOSE: To evaluate the accuracy of Current Procedural Terminology (CPT) coding for interventional radiology procedures and the associated professional economic impact when coding is performed by operating physicians. METHODS: Procedure reports and physician charge sheets were obtained for 549 interventional radiology encounters performed by 62 physicians at 23 hospitals and analyzed for appropriate CPT code use. Physician-selected CPT codes were reviewed by experts, who determined correct coding by consensus. Physician coding errors and professional relative value unit (RVU) impact were analyzed. Expert discordance and associated RVU impact were similarly evaluated. RESULTS: Physicians correctly coded only 242 of 549 IR cases (44%). The overall professional RVU impact of their errors was +4.2% (overcoding). Physician coding was correct least frequently for complex arterial interventions (15 of 53, 28%) and dialysis access interventions (16 of 54, 30%) and correct most frequently for less code-intensive drainage (19 of 31, 61%) and biopsy procedures (35 of 47, 74%). Experts were initially concordant in 497 of 549 cases (91%), with only a minimal tendency (+0.3% RVU) toward overcoding. Expert coding differences were explained by simple code oversights (28 of 52, 54%), coding guideline ambiguity (15 of 52, 29%), and physician documentation ambiguity (9 of 52, 17%). CONCLUSION: When interventionalists code their own procedures, CPT errors are common, but the associated RVU impact is small. Given the consequences of incorrect coding, physician-assigned CPT codes warrant review by experienced coders before claims submission. Although radiology practices should strive for perfect coding, expert discordance suggests that this goal is unattainable but less elusive than for nonradiology services. PMID- 17411694 TI - Electronic publishing in radiology: beginnings, current status, and expanding horizons. AB - Electronic publishing in radiology began in the 1980s and gathered momentum as use of the personal computer and subsequently the World Wide Web became commonplace. The ease of access and wide distribution that the Internet affords have presented both experts and lay users with the challenge of distinguishing reliable from unreliable material. In the field of radiology, peer-reviewed journals, the sine qua non of reliability in the scientific realm, began to appear in online versions in 1988, and now, nearly all such journals currently have online versions. Electronic versions of textbooks and textbook-like material have become more common, although the vast majority of publishers' catalogs are traditional print books. Electronic material is more common in the fields of internal, emergency, and family medicine than in the field of diagnostic radiology. The latest publishing medium is the personal digital assistant. Challenges that face electronic publishing include the issues of permanent archiving, citation, and indexing. Peer review seems to be the best method for distinguishing reliable from unreliable information. The use of Web technology has improved the logistics of peer review, and some journals have begun posting peer reviews themselves on the Web alongside peer-reviewed articles. Electronic publishing has changed the manner in which radiologists obtain information, providing wider, more immediate access. PMID- 17411695 TI - Keeping it safe: MRI site design, operations, and surveillance at an extended university health system. AB - Magnetic resonance imaging (MRI) safety has attracted increasing attention both within the radiology community and in general. Although much of this attention has been spurred by high-profile accidents, the wide distribution of increasingly powerful MRI technology makes safety a paramount concern. While the ACR has sponsored an overarching policy statement regarding MRI safety (The ACR White Paper on MR Safety), and many articles have reported specific hazards and evaluated myriad devices, this paper describes in detail the practical implementation of safe MRI practice at a large medical center. The implementation focuses on 3 key areas: (1) site access restriction, (2) personnel management, and (3) equipment. In addition, a comprehensive and proactive program for MRI safety surveillance and preemptive action is presented. This last step is key to maintaining the effectiveness of an MRI safety program. PMID- 17411696 TI - Beating them or joining them: your radiology group's path to the future. AB - Radiologists and hospital and health system (HHS) administrators and trustees are facing significant threats and opportunities in the rapidly changing health care environment. Although multiple response alternatives exist for both groups, some of the options seem more likely than others to lead to long-term success. The choices available range from cooperation to competition, from bargaining to blaming, from offering hope to providing criticism, from standing together to standing apart, and from focusing on beating them to investing in joining them for the benefit of all. Many physicians and HHS administrators and trustees acknowledge that their relationship with each other is central to their long-term success, as well as the interests of patients and their communities. This article discusses the major forces for change faced by radiologists and HHS administrators and trustees. It examines the collaborative and competitive responses available to each, provides a review of the typical consequences of these responses, and identifies the assumptions underlying the major alternatives. PMID- 17411697 TI - Medical entrepreneurism: the current opportunity in America. AB - This article discusses both the current climate for entrepreneurial activity in the fields of biotechnology, health care services, and medical devices as well as key ideas in the process of successful innovation. Basic issues related to the nature of new medical ventures and its importance in the U.S. economy are discussed. A stepwise overview of the process of innovation is provided, starting from the initial idea, through the early and middle stages of growth, and on to an initial public offering or other alternative harvest strategy. The roles of financing sources in generating health care entrepreneurial activity are explored, and the advantages and disadvantages of each are discussed. The article focuses on venture capital investment because of its pivotal role in high-profile successes; however, alternative forms of financing are also covered as appropriate to each stage. In addition, critical nonfinancial issues that affect the success of new enterprises, such as intellectual property protection and the creation of management teams for young companies, are also covered. The magnitude of current investment in the domestic biomedical field is addressed, and future prospects for American medical innovation are briefly discussed. PMID- 17411698 TI - Can breast and cervical cancer screening visits be used to enhance colorectal cancer screening? AB - PURPOSE: Despite high acceptance levels of mammography and cervical cancer screening by U.S. women, adherence with colorectal cancer screening remains suboptimal. A better understanding of the relationship among cancer screening behaviors by women may provide insight into interventions to enhance colorectal cancer screening. METHODS: Women 50 years and older who participated in the 2000 Behavioral Risk Factors Surveillance Survey and lived in one of the five states that administered the colorectal cancer module (Colorado, Illinois, Massachusetts, Ohio, and Utah) were queried regarding cancer screening patterns. Predictors of colorectal cancer screening were determined using multivariate analysis from sociodemographic data and non-colorectal cancer screening adherence rates (based on American Cancer Society guidelines). RESULTS: Among the 1300 colorectal cancer module respondents, cancer screening adherence was significantly less for colorectal cancer (24.9%) compared with cervical cancer (57.2%) or breast cancer (78.6%). In multivariate analysis, increasing age, health insurance, adherence with cervical cancer screening (adjusted odds ratio [OR] 2.09, p < 0.01) and adherence with breast cancer screening (adjusted OR 1.89, p < 0.01) were independent predictors of colorectal cancer screening. Participants who adhered to both mammography and Pap smear guidelines were significantly more likely to adhere to colorectal cancer screening compared with women who adhered to either screening test alone (adjusted OR 1.88, p < 0.001). CONCLUSIONS: Women adherent to mammography and cervical cancer screening guidelines were significantly more likely to undergo colorectal cancer screening than those who were not adherent, although colorectal cancer acceptance in the adherent group was still suboptimal. Because psychological barriers to colorectal cancer screening exist, non-colorectal cancer screening visits that women already readily accept potentially represent a setting (or "teachable moment") for the delivery of education and behavior-related interventions aimed at reducing the burden of colorectal cancer. PMID- 17411699 TI - National practitioner data bank: information on physicians. PMID- 17411700 TI - Image compression in your PACS: should you do it? What are the issues? PMID- 17411701 TI - Radiology job sharing: improving quality of life and maximizing resources for radiology groups. PMID- 17411702 TI - Radiation risk from airline travel. PMID- 17411703 TI - A resident's perspective on self-referral. PMID- 17411704 TI - Chapter resident education. PMID- 17411705 TI - Geese and golden eggs. PMID- 17411706 TI - Self-referral: the time for whining is over. PMID- 17411707 TI - Mammography and the risks of engagement. PMID- 17411708 TI - Strategies for attracting residents to understaffed radiologic specialties: the case of pediatric radiology. PMID- 17411709 TI - Revenue-based compensation. PMID- 17411710 TI - Turf wars in radiology: possible remedies for self-referral that could be taken by federal or state governments and payers. AB - The time seems to be ripe for reforms that will address the rapid growth of self referral in diagnostic imaging by nonradiologist physicians. The authors present a possible course of action containing six elements that federal or state governments and/or payers could take that would assist them in their efforts to control this ubiquitous problem: (1) a legislative ban on self-referral for certain types of imaging, (2) mandatory accreditation and/or site inspections of all imaging facilities, (3) the limitation of imaging privileges among nonradiologists, (4) the required precertification of all self-referred imaging examinations, (5) the auditing of records of physicians who self-refer, and (6) certificate-of-need laws. PMID- 17411711 TI - Experience-based consulting: the value proposition. AB - Consulting is a profession universally accepted and well entrenched throughout the business world. Whether it is providing objective analysis, supplying a specific expertise, managing a project, or simply adding extra manpower, consultants can add value. However, what are the attributes of a good consultant? In health care, with the rapid pace of emerging technologies, economic intricacies, and the complexities of clinical care, hands-on experience is the key. Recognizing the power of consultants with hands-on experience, the Department of Radiology at Massachusetts General Hospital launched the Radiology Consulting Group, an "experience-based" model for consulting that may potentially shift the profession's paradigm. PMID- 17411712 TI - Electronic publishing in radiology: economics and the future. AB - Scholarly publishing is a large market involving thousands of peer-reviewed journals but a decreasing number of publishers. An economic model can be described in which authors give their work to publishers who then sell access to this work. Because each published article is a unique work with few if any substitutes, publishers have some degree of monopoly power and can price their products accordingly. The advent of desktop publishing using personal computers made it possible for individuals to publish material without publishers, an activity that gained momentum when the publishing medium shifted from paper to electronic, and from electronic publishing to the Internet. This activity destabilized the industry, and in the rush to gain market share by providing free content, unsustainable business models were created. Scholarly publishing is now dominated by a small number of multinational corporations that acquired many smaller publishing operations. As these companies have exercised their monopoly power, an open access movement has gained traction in which authors (or their institutions) initially pay for publication, but readers have free and open access to the published articles. This movement is in diametric opposition to the commercial publishing model, and it remains to be seen whether and how well the two can coexist in the future. PMID- 17411713 TI - Ten filmless years and ten lessons: a 10th-anniversary retrospective from the Baltimore VA Medical Center. AB - The authors review a decade's experience in the nation's first filmless radiology department and outline the challenges and rewards of the transition. They summarize their experience with 10 cautionary and informative lessons on making the process more successful, more efficient, and less stressful. A number of possible avenues of new research and assessment on the effects of filmless operation on radiologists, imaging staff, referring clinicians, and patients are highlighted. PMID- 17411714 TI - The private-practice perspective of the manpower crisis in radiology: greener pastures? AB - Rising consumer expectations and a rapidly aging population point to a long-term shortage of all physicians, including radiologists. While attention has been drawn to the escalating manpower crisis in academic radiology departments, the private-practice perspective has been generally overlooked. Although clinical workloads and income are higher in private practice, studies have shown higher satisfaction levels (likely because of a greater variety of work) among academic radiologists. As the distinction between community and teaching hospitals has become increasingly blurred, there is now considerable overlap in the skill sets, sources of job satisfaction, and stresses that are encountered in both practice settings. Perhaps more than at any time in the recent past, diagnostic radiologists in academic and private practice share more in common than any perceived differences. Both groups must work together in concert with the ACR to address the growing manpower shortage, as well as the other challenges that confront diagnostic radiology at the beginning of the 21st century. PMID- 17411715 TI - Resolving the technologist shortage: a summary of ACR and ASRT efforts to meet the increasing demand for RTs. AB - Demand for radiologic technologists in the United States greatly outstripped supply throughout the late 1990s and peaked in 2000, when vacancy rates for radiologic technologists reached an average of 18% nationwide. To combat the shortage, the ACR and the American Society of Radiologic Technologists (ASRT) launched a series of aggressive recruitment and retention initiatives designed to boost the number of technologists. The campaigns have resulted in rising enrollments in educational programs, greater numbers of graduates and new technologists, and the expansion of the career ladder. As a result, the national vacancy rate for radiologic technologists had dropped to 12% by the end of 2003. This article reviews the radiologic technologists personnel shortage at the turn of the century, describes efforts taken by the ACR and the ASRT to ameliorate the shortage, and examines the future of the radiologic technology workforce. PMID- 17411716 TI - Ensuring the radiologist workforce: opportunities during education. AB - The authors conducted a literature review to better understand the current trends in the medical workforce and, specifically, the effect these have had in radiology. The cyclic nature of the radiology workforce supply has paralleled similar experiences in the general medical profession and other specialties. A review of proven education and training initiatives demonstrates positive workforce effects in particular circumstances. The adaptation of proven education initiatives should enhance the future radiology workforce in both private and academic settings. PMID- 17411717 TI - Implementing digital quality control in a breast center. AB - Breast imaging centers in the United States are currently transitioning from traditional screen-film mammography to full-field digital mammography. Quality control for full-field digital mammography is more difficult, time consuming, and expensive than for traditional screen-film mammography. Full-field digital mammography quality control is more precise than screen-film mammography quality control at identifying noncompliant components. As per U.S. Food and Drug Administration requirements, each manufacturer of full-field digital mammography equipment designs and establishes its own quality control process. PMID- 17411718 TI - The appropriateness of employing imaging screening technologies: report of the methods committee of the ACR task force on screening technologies. AB - Screening for disease using imaging technologies is a growing phenomenon. For some applications (e.g., mammography and breast cancer), there is solid evidence that imaging screening reduces disease-specific mortality. However, for other applications, the public interest in imaging screening is progressing despite a lack of valid scientific evidence that such screening improves the public's health. Clinical trials promise to eventually shed light on the value of imaging screening, but the results of these trials will not be known for several years. In the meantime, physicians and the public need guidance on whether specific imaging screening examinations are appropriate. The ACR charged the Methods Committee of its Task Force on Screening Technologies to advise the organization on how it could best use available evidence to make public recommendations on imaging screening technologies. This article is the outcome of the committee's deliberations, including recommendations for the method of arriving at a recommendation, the approach to dissemination, and to whom the recommendations should be targeted. PMID- 17411719 TI - Update on the National Cancer Institute's clinical trials cooperative group program. PMID- 17411720 TI - International radiology quality network. PMID- 17411721 TI - MRI site planning adventures. PMID- 17411722 TI - Dictating for dollars (and compliance, too). PMID- 17411723 TI - Outpatient imaging center ventures: a third-party, for-profit perspective. PMID- 17411724 TI - Teaching residents the art of defensive radiology. PMID- 17411725 TI - Appropriateness criteria: the double-edged sword. PMID- 17411726 TI - Who gets paid with "pay-for-performance"? PMID- 17411727 TI - Radiology in the era of accountability: perfect storm or new dawn? PMID- 17411728 TI - Trusting the student: learner-centered education. PMID- 17411729 TI - Health care inflation and high-tech medicine: a new look. PMID- 17411730 TI - Is smaller better? PMID- 17411731 TI - Expert or fact witness: is there a legal obligation to testify? PMID- 17411732 TI - Report of the ACR task force on medical liability reform in radiology. AB - The ACR Board of Chancellors commissioned a task force to investigate the issue of needed reforms in medical liability as it relates to the specialty of radiology. This article relates the task force deliberations and recommendations. PMID- 17411733 TI - Planning and developing a diagnostic imaging center. AB - The convergence of an aging population, new technology, and an increasing number of clinical applications and reimbursements has created a favorable environment for ambulatory imaging centers. From a patient's perspective, features such as easy access, parking, appointment availability, and a setting that caters to outpatients are attractive compared with the hospital environment. Before embarking on a venture, the execution of a careful, thorough planning process, from the inception of the idea to the opening, is vital to success. This article provides guidance on the process by discussing the process of determining whether a project is feasible; developing a business plan that will measure potential success; locating, financing, and planning space; contractor selection and project management; and planning operations. PMID- 17411734 TI - Multipayer provider agreements: the gift that keeps on giving (and giving, and giving...). AB - This article discusses provider agreements that can be accessed by various different payers rather than solely by the single parties that originated the agreements. In recent years, these "multipayer agreements" have been used both more widely and for wider purposes than in the past. The application of multipayer agreements is becoming more complex and fraught with errors, which often disadvantage imaging practices. Even without error, the broader use of multipayer agreements can have a significant and often unrecognized adverse impact on reimbursement for imaging services. This article describes multipayer agreements in detail, including why they were originally developed, why payers use multipayer agreements, and why their use has expanded recently, notably to national insurers. Furthermore, this description identifies two types of multipayer agreements. A "keychain" agreement allows multipayer plans to unilaterally add new payers to existing provider agreements, each of which becomes entitled to the discounts and other benefits conferred by the base agreement. A "power-of-attorney" agreement allows multipayer plans to effectively bind providers to new provider agreements unilaterally. Three examples of issues confronted by imaging practices in the course of administering existing multipayer agreements are described in some detail. Last, the article provides several suggestions for evaluating and containing an imaging practice's potential exposure to multipayer agreements. PMID- 17411735 TI - Who gets paid for diagnostic imaging, and how much? AB - PURPOSES: To determine the apportionment of Medicare Part B payments for noninvasive diagnostic imaging among radiologists and other specialists and to ascertain the trends in these payments over a recent 5-year period. METHODS AND MATERIALS: Using the nationwide Medicare Part B databases for 1997 through 2002, all payments were calculated for physician services for noninvasive diagnostic imaging codes in the 70,000 Current Procedural Terminology, Version 4 series and those codes in the 90,000 series pertaining to echocardiography and vascular ultrasound. Imaging studies were grouped into 26 different categories depending on type and modality. The 108 Medicare physician specialty codes were used to group payments into 20 different specialist categories. The percentage shares for each specialty group in 1997 and 2002 were calculated. Percentage changes in payments between 1997 and 2002 were determined. RESULTS: In 2002, Medicare Part B paid almost $7.7 billion for noninvasive diagnostic imaging services. Radiologists received 48.3% of these payments, and cardiologists received 22.8%. Between 1997 and 2002, overall payments to physicians for noninvasive diagnostic imaging rose by 78%. Payments to radiologists increased by 65%, while payments to cardiologists increased by 105%. Primary-care physicians received 8.3% of 2002 payments, while 10.2% went to independent diagnostic testing facilities or multispecialty groups. For the latter two entities, the specialties of the physician providers could not be determined. CONCLUSION: Radiologists receive only about half of all Medicare Part B reimbursements for noninvasive diagnostic imaging. Cardiologists receive almost one-quarter. Payments to cardiologists are rising more rapidly then to any other specialty group and considerably more rapidly than payments to radiologists. PMID- 17411736 TI - The current status of full-field digital mammography quality control. AB - U.S. Food and Drug Administration (FDA) regulations require that medical physicists and technologists adhere to the procedures, testing frequencies, and performance criteria outlined in the quality control (QC) manuals provided by the manufacturers of full-field digital mammography (FFDM) systems. As a result of this requirement, QC for digital equipment has become both complicated and inconsistent. The objective of this study was to compare the current QC protocols for the tests to be performed by medical physicists for three commercially available FFDM systems (the Senographe 2000D, the SenoScan, and the Selenia). Although not yet approved by the FDA, the 5000 MA was also included because of its unique design and extensive use in a major ongoing clinical research trial involving the other three FFDM systems. It was found that currently, a high degree of discord exists between the FFDM QC protocols of the early commercial systems, with wide variation in opinion with respect to the minimum performance criteria and testing intervals and with respect to which tests are necessary and what quantities should be measured. Additionally, very few digital detector specific tests are currently in place. Although technological differences between the early FFDM systems prohibit a strictly uniform set of QC procedures and performance guidelines, a much greater degree of consensus among manufacturers is desirable as FFDM continues to gain clinical presence during the next few years. PMID- 17411737 TI - Paying for performance in diagnostic imaging: current challenges and future prospects. AB - As concerns regarding medical errors and patient safety continue to escalate, policy makers and purchasers of care are developing novel methods to improve health care quality. One strategy for encouraging quality improvement uses financial incentives to reward physicians and hospitals for meeting or exceeding benchmarks of quality. This model, known as "paying for performance," is currently being implemented in a number of health care markets. Although most of these programs focus on primary care physicians and hospital care, it is only a matter of time until pay-for-performance incentives will be designed for diagnostic imaging groups. This article describes the pay-for-performance model and discusses its implications for the field of radiology. PMID- 17411738 TI - Imaging to optimally stage lung cancer: conventional modalities and PET/CT. AB - Accurately staging patients with lung cancer is important in determining treatment options and prognoses. Staging allows the distinction of patients who are candidates for surgical resection from those with inoperable disease who may be treated with chemotherapy and/or radiation therapy. Conventional imaging plays an essential role in the noninvasive and invasive methods of the evaluation and staging of patients with non-small-cell lung cancer (NSCLC). Imaging modalities used for staging include chest radiography, chest computed tomography (CT), abdominal CT, brain CT or magnetic resonance imaging, bone scans, and (18)F-2 deoxy-d-glucose positron emission tomography (PET). Recently, PET/CT, the integration of the functional data of PET with the anatomic data of CT, has emerged as a modality to potentially change the way patients are evaluated. This article reviews current recommendations regarding the staging of patients with NSCLC and addresses the role of PET/CT. PMID- 17411739 TI - The use of an interactive computerized daily schedule in a busy interventional radiology practice increases efficiency. AB - PURPOSE: The effect on efficiency of patient care in a busy academic interventional radiology practice was studied by the analysis of procedure times both before and after the implementation of a computerized interactive daily schedule. Procedure start and end times were retrospectively collected from the department's quality assurance database for two identical 6-month periods, representing the time before and after the deployment of the software. The delay in the start of the first case, the time between cases, and the time required to complete each day's work were compared for the two periods. RESULTS: The average time of delay between cases was reduced after the implementation of the software (p < 0.025). More total cases were performed during the period of time after the implementation of the software, resulting in a greater work relative value unit production. Although the average number of cases performed per day was greater after the software was in use (p < 0.03), the average amount of time required to complete the day's work was not significantly changed (p = 0.08). There was no apparent effect on the average delay of the start of the day's first case (p = 0.34). CONCLUSION: The use of a computerized interactive daily schedule has a positive effect on departmental efficiency by allowing more cases to be performed without lengthening the workday. PMID- 17411740 TI - The polity of academic medicine: a critical analysis of autocratic governance. AB - How should academic radiology departments be governed? This question has rarely been directly addressed in the radiology literature. The dominant model of administration in present-day academic departments differs from that typically seen in private group practices. Whereas private group practices tend to follow a democratic model whereby key decisions must be supported by a majority of the partners, in academic institutions, medical school deans and department chairs generally possess great latitude in strategic and operational decision making. This article considers arguments for and against "top-down" governance in academia. The rationale supporting this form of governance is weak, and the best evidence from the fields of management and organizational behavior suggests it may in fact be detrimental. PMID- 17411741 TI - To compress or not to compress: a compressed debate. PMID- 17411742 TI - Radiology peer review as an opportunity to reduce errors and improve patient care. PMID- 17411743 TI - Alice in radiation protection land. PMID- 17411744 TI - The case for the three-year residency. PMID- 17411745 TI - Radiologist-patient contact: a different perspective. PMID- 17411748 TI - Introduction to molecular imaging. PMID- 17411749 TI - Clinical molecular imaging. AB - This review summarizes the rapidly growing field of molecular imaging, the spatially localized and/or temporally resolved sensing of molecular and cellular processes in vivo. Molecular imaging is used to map the anatomic locations of specific molecules of interest within living tissue and has enormous potential as a powerful means to diagnose and monitor disease. Molecular imaging agents comprise a targeting component that confers localization and a component that enables external detectability with an imaging modality, such as PET, SPECT, MRI, optical, and ultrasound. The advantages and disadvantages of each of these modalities are discussed in regard to spatial resolution, temporal resolution, sensitivity, and cost. Molecular imaging agents can be divided into three categories, Type A, which bind directly to a target molecule, Type B, which are accumulated by molecular or cellular activity by the target, and Type C, which are undetectable when injected but can be imaged after they are activated by the target. The current status of clinical molecular imaging agents is presented as well as examples of some preclinical applications. The value of molecular imaging is illustrated by some examples for diseases such as cancer, neurological and psychiatric disorders, cardiovascular disease, infection and inflammation, and the monitoring of gene therapy and stem cell therapy. PMID- 17411751 TI - Valid clinical research and reimbursement for what we do. PMID- 17411752 TI - Why advances in imaging technology are good for radiology, patients, and the bottom line. PMID- 17411753 TI - Developing a national medical student curriculum in radiology. PMID- 17411754 TI - A surgeon leads the Senate. PMID- 17411755 TI - Diagnostic radiology reporting and communication: the ACR guideline. AB - The ACR adopted its "Guideline for Communication: Diagnostic Radiology" in 1991. Since its adoption, the guideline has been the subject of considerable discussion and controversy. In response to more than a decade of debate, the ACR appointed a task force in the summer of 2003 to research and analyze claims and litigation decisions that have been related to the communication or reporting of imaging studies by radiologists. Furthermore, the task force was charged with making recommendations regarding the status and impact of the existing communication guideline. The only specific directions to the task force were to take into account the ACR's motto, "Quality is our image," in the recognition that communication plays an essential role in safety and quality. The task force consulted outside legal counsel, reviewed claims data from many sources, and performed a survey of the ACR's membership. Furthermore, the task force was divided into four working groups to focus on the data and make specific recommendations. The products of the working groups were assembled into a final report that was presented to the ACR Board of Chancellors in the winter of 2004. This report, including five recommendations, and a draft for a new communication guideline were presented to the ACR Council at the annual meeting in May 2004. PMID- 17411756 TI - Maintenance of certification. AB - Maintenance of certification (MOC) recognizes that in addition to medical knowledge, several essential elements involved in delivering quality care must be developed and maintained throughout one's career. The MOC process is designed to facilitate and document the professional development of each American Board of Radiology (ABR) diplomate through its focus on the essential elements of quality care in diagnostic radiology, its subspecialties, radiation oncology, and radiologic physics. The initial elements of the ABR's MOC program have been developed in accord with guidelines of the American Board of Medical Specialties. Further details will be developed as the process evolves. PMID- 17411757 TI - The Blue Cross Blue Shield Association Technology Evaluation Center: how we evaluate radiology technologies. AB - Evidence-based technology assessment can help answer critical questions concerning the safety, effectiveness, and appropriate uses of medical technologies. This practice can be used to avoid the promotion of ineffective technologies and the premature diffusion of technologies that have not been demonstrated to improve patient-oriented health outcomes, both of which draw resources from effective and appropriate medical care. This article describes the process of such evaluation as undertaken by the Blue Cross Blue Shield Association Technology Evaluation Center. The key components of the assessment process are described, including the problem formulation and evaluation of study quality, as well as the process by which the available evidence is judged against the five Technology Evaluation Center criteria. PMID- 17411758 TI - The changing roles of radiologists, cardiologists, and vascular surgeons in percutaneous peripheral arterial interventions during a recent five-year interval. AB - PURPOSE: This study aimed to determine the relative roles of radiologists, cardiologists, vascular surgeons, and other physicians in performing percutaneous peripheral arterial interventions and how these roles have changed over a recent 5-year period. METHODS AND MATERIALS: The authors reviewed the Medicare Part B fee-for-service databases between 1997 and 2002 for the Current Procedural Terminology (4th ed.) (CPT-4) surgical procedure codes for percutaneous transluminal angioplasty (PTA) of noncardiac peripheral arteries (six codes), the transcatheter placement of noncardiac intravascular stents (two codes), and endovascular aortic stent graft placement (six codes). Using the Medicare physician specialty codes, procedure volume in each CPT-4 code was determined for radiologists, cardiologists, vascular surgeons, and other physicians. Percentage changes from 1997 to 2002 were calculated for PTA and intravascular stent placement procedures. RESULTS: Between 1997 and 2002, the total Medicare procedure volume in the eight procedure codes relating to PTA and stent placement increased by 95%. In 2002, radiologists performed 72,657 of these procedures, cardiologists 62,901, vascular surgeons 17,895, and other physicians 19,666. Over the 5-year interval, procedure volume among radiologists increased 29%, among cardiologists by 181%, among vascular surgeons by 398%, and among other physicians by 195%. Radiologists' share in the total pool of procedures in 2002 was 42.0% (down from 63.3% in 1997), cardiologists' 36.3% (up from 25.2% in 1997), vascular surgeons' 10.3% (up from 4.0% in 1997), and other physicians' 11.4% (up from 7.5% in 1997). Trend data were not available for endovascular aortic stent graft procedures. CONCLUSION: Between 1997 and 2002, procedure volume in percutaneous peripheral arterial interventions grew at faster rates among cardiologists, vascular surgeons, and other physicians than it did among radiologists. As a result, radiologists' share of this market declined during the interval. However, procedure volume among radiologists continued to grow over the 5 years, and in 2002, they still had the largest share among the four physician specialty groups. Thus, despite the erosion, interventional radiologists still maintain a strong position in this rapidly growing field. PMID- 17411759 TI - The future of imaging screening: proceedings of the Fourth Annual ACR FORUM. AB - The FORUM is an ACR planning activity focused on a specific topic deemed to be of long-range importance to the college. A select panel of multidisciplinary experts met in 2004 to consider the ramifications of imaging screening from diverse perspectives. Considerations included the nature of screening for disease, its technological issues, and biases associated with the perceived success of screening; potential technologies and target diseases; business and economics; insurance coverage; ethics and the law; how imaging screening could be paired with nonimaging screening; and how the lay public perceives screening. The FORUM participants considered how imaging screening might develop over the next 5 to 10 years and made recommendations to the ACR on actions it might take to benefit both the specialty and patients. PMID- 17411760 TI - A minority view to the ACR FORUM: advancing the cause of imaging screening. PMID- 17411761 TI - Factors to consider in the revision of educational objectives for medical students in radiology clerkships. AB - PURPOSE: To maintain accreditation by the Liaison Committee on Medical Education and to form up-to-date educational objectives for medical students rotating through radiology clerkships, the authors investigated the influencing factors from instructors', students', and the medical community's perspectives in the setting of traditional radiology medical student education. METHOD: Radiology instructors' perspectives were studied via literature, experience, and consensus. The medical community's perspectives were studied via literature, most specifically the recommendations of the Liason Committee on Medical Education. Medical students' perspectives were studied via a survey questionnaire eliciting the goals of the students as their clerkships began. RESULTS: Radiology instructors' goals for radiology clerkships centered on anatomy, pathology, technology, and foundation. The medical community's goals for radiology clerkships centered on history taking, physical examination, procedures, diagnosis, management, prevention, data analysis, communication, professionalism, cultural competence, careers in medicine, and models of health care delivery. Thirteen medical students were surveyed, and 13 responded (100%). Medical students' goals for their radiology clerkships centered on appropriateness, science, film interpretation, and confidence. CONCLUSION: The objectives for radiology clerkships in medical schools should be reevaluated and rewritten frequently. The formulation of the optimal objectives should be multifactorial, including the input of medical students, radiology instructors, and the medical community. These factors should be incorporated in the setting of traditional educational values with special consideration to current medical students' needs. The authors have provided the objectives that they recently adopted, which they feel appropriately address the currently important issues. PMID- 17411762 TI - The ACR appropriateness criteria: translation to practice and research. AB - The ACR Appropriateness Criteria (ACRAC) are now just about a decade old. Along with a continued commitment to update and add to these guidelines, the author advocates that the ACR undertake the task of converting the current text-based contents into a comprehensive, multipurpose relational database. A candidate structure for such a database is described, and several uses for it are articulated. The accompanying policy discussion advocates efforts to widely disseminate the resulting ACRAC database for use by payers in setting reimbursement and by hospitals in radiology order entry software. Also described are some ways that the ACRAC, formatted as a database, can be used for health services research. PMID- 17411763 TI - Turf wars in radiology: the battle for peripheral vascular interventions. AB - Interventional radiologists in many hospitals are involved in confrontations with cardiologists and vascular surgeons over who should be allowed to perform percutaneous noncardiac peripheral vascular interventions. There are valid reasons why radiologists should be the ones doing these procedures: first, because in any given hospital, radiologists are generally the physicians with the best training and most experience, and second, because they are generally not in a position to self-refer and will therefore be able to help keep utilization under control. If cardiologists or vascular surgeons request vascular interventional privileges at your hospital, there are steps you can take to see if they are properly qualified. If they are granted privileges, there are other steps you can take to ensure that high standards of patient care are maintained. The authors also present some discussion of how interventional radiologists can position themselves to either compete with or collaborate with the other clinical services. Throughout any confrontations that might occur, radiologists should stress that patients undergoing these procedures deserve the best possible care, which means that they should be performed by those physicians on the hospital staff who are the most knowledgeable and the least likely to commit medical errors. PMID- 17411764 TI - Publication rate of abstracts from the annual ASTRO meeting: comparison with other organizations. AB - PURPOSE: One of the explicit goals of the American Society of Therapeutic Radiology and Oncology (ASTRO) is to promote research and disseminate research results. In the past few years, ASTRO has required that manuscripts be submitted for publication for all papers accepted for oral presentation at its annual meeting. The purpose of this study was to determine the publication rate of abstracts accepted for oral presentation at ASTRO's 1999, 2000, and 2001 annual meetings. MATERIALS AND METHODS: The authors reviewed the proceedings of ASTRO's annual meetings in 1999, 2000, and 2001 to identify all abstracts accepted for oral presentation. The following information was collected: year of presentation, study design (phase I or II, phase III, or retrospective), country of origin (domestic or foreign), abstract category (clinical or nonclinical), disease site (if applicable), publication (yes or no), publication date, and publishing journal. A computer-based search using Medline was used to determine whether the full publication of each abstract had occurred. The computer search included publication up to November 1, 2003. RESULTS: The publication rate was 56% (452 of 802). There was no difference in publication rate according to country of origin (domestic 56%, foreign 57%; p = NS), abstract category (clinical 59%, nonclinical 48%; p = NS), or study design. Half of the published abstracts were published within 1 year of the meeting, and 90% were published within 2 years. The 452 publications were distributed among 54 different journals. The majority of papers were published in the International Journal of Radiation Oncology, Biology and Physics (62%), followed by the Journal of Clinical Oncology (8%) and Radiotherapy and Oncology (3%). CONCLUSIONS: Slightly more than one-half of the abstracts accepted for oral presentation at the annual ASTRO meeting are published within 2 years. This rate is similar to those of other specialties and suggests that ASTRO is succeeding in its mission to promote and disseminate research. PMID- 17411765 TI - The golden rule. PMID- 17411766 TI - New rules, new codes, and the fall from grace. PMID- 17411767 TI - Follow the money! PMID- 17411768 TI - MRI: worth a Nobel Prize? PMID- 17411769 TI - What's new in brachytherapy? PMID- 17411770 TI - The vital role of residents and fellows. PMID- 17411771 TI - Re: "Mammography and the risks of engagement". PMID- 17411772 TI - Re: "Patient radiation ID cards". PMID- 17411773 TI - Re: "Evidence-based medicine: ready for prime time?". PMID- 17411774 TI - The viability of our specialty: the designated physician imager. PMID- 17411775 TI - International teleradiology and new workforce expectations. PMID- 17411776 TI - Nontechnical aspects of PACS. PMID- 17411777 TI - Is there a future for real-time ultrasound scanning by radiologists? PMID- 17411778 TI - Health care in America: sustainable path or collision course? PMID- 17411779 TI - Medical oncology, surgical oncology, radiation oncology...interventional oncology? PMID- 17411780 TI - Report of the ACR task force on international teleradiology. AB - Telemedicine is becoming an increasingly important tool in the practice of medicine throughout the world. For radiologists, telemedicine translates to teleradiology. Because an increasing amount of imaging is now archived in a digital format, and with the application of more powerful computers in radiology, digital image transmission between display stations is becoming commonplace. The ability to move large diagnostic image data sets to display stations anywhere in the world using the Internet and other high-speed data links is solving some problems and creating others. Medicine and radiology will be challenged in many ways by the issues created from the application of this burgeoning technology. Our task force was charged with investigating the evolving practice of international teleradiology and with developing a pubic statement to be adopted by the ACR Council (). This white paper is our effort to define those issues we believe to be most pertinent to international teleradiology as we know them today. Will these issues be changing? Certainly. For some facets of the issue, there are currently more questions than answers. We describe several scenarios that we believe are acceptable practices of international teleradiology as well as some that are not. We believe that much will be written about international teleradiology in the future as the issues of credentialing, quality assurance, licensure, American Board of Radiology certification, the maintenance of certification, jurisdictional and medical liability issues, patient privacy, fraud and medical ethics are more precisely defined and shaped by state and federal legislation and medical jurisprudence. This white paper is our assessment of what we believe to be the major challenges that exist as of this writing. PMID- 17411781 TI - Expert testimony: a perspective from the trenches. AB - The controversy over the role of the "expert" in medical litigation is in some ways the product of semantic confusion. Many physicians mistake the legal definition of an expert with that of ordinary usage, further fueling the resentment of doctors who testify against other doctors. Legally, however, an expert is simply one qualified to offer an opinion. In ruling on the admissibility of expert testimony, a court determines whether the proffered expert has the minimal qualifications necessary to have an opinion. An expert, then, does not have to have the right opinion; rather, he or she has the right to have an opinion. The bar is thus set quite low. In medical malpractice litigation, the threshold of admissibility is usually met by board certification in the same specialty as the defendant. Although expert testimony, as defined above, is necessary in most medical malpractice cases, that need is obviated when negligence is so obvious that the law presumes that a lay jury can discover it. Abuses of expert testimony are real. They are, however, in some part the consequence of some of the best doctors' refusal to assist the victims of malpractice, creating a vacuum that the less scrupulous fill. PMID- 17411782 TI - Expert testimony: a contrary perspective from the trenches. PMID- 17411783 TI - The future workforce in academic radiology: gender, generational, and cultural influences. AB - The acute shortage of academic radiologists in the United States, and in fact across the world, is a multifactorial issue. A review of the recent literature suggests a number of influences that discourage radiology residents from remaining in academics, including cultural and generational values and their gender roles in our society. This article discusses the influence of gender, generational, and cultural issues on the current and future workforce in academic radiology. PMID- 17411784 TI - The fundamentals of the US Medicare physician reimbursement process. AB - The Medicare program, enacted in 1965, is a federally funded health care coverage plan for people aged 65 years and older, for those who are disabled, and for those needing renal dialysis or kidney transplants for the treatment of end-stage renal disease. Today, nearly 40 million Americans rely on Medicare for their health care services. The purpose of the Medicare program is to increase access to quality care for the elderly while maintaining a financially viable federal fund from which health care reimbursements can be appropriately and efficiently allocated to health care providers. This paper has three main objectives: (1) introduce the functioning of the Medicare payment system, (2) explain in more detail how the program currently reimburses physicians, and (3) discuss the current challenges facing the physician reimbursement system. PMID- 17411785 TI - Effective radiology teams. AB - Teams can be very helpful in quality improvement, the service recovery process, and reengineering. Teams can increase organizational communication, quality, productivity, and morale. However, not all teams work well, and some can waste time, cause resentment, and be a net negative for an organization. This article summarizes some of the steps in getting a team to a good start, keeping it running well, having effective meetings, and correcting some common problems. PMID- 17411786 TI - A framework for improving radiology reporting. AB - The interpretative reports rendered by radiologists are the only tangible manifestation of their expertise, training, and experience. These documents are very often the primary means by which radiologists provide patient care. Radiology reports are extremely variable in form, content, and quality. The authors propose a framework for conceptualizing the reporting process and how it might be improved. This consists of standard language, a structured format, and consistent content. These attributes will be realized by modifying the clinical reporting process, including the creation, storage, transmission, and review of interpretative documents. The authors also point out that changes in training and evaluation must be a part of the process, because they are complementary to purely technical solutions. PMID- 17411787 TI - Web-based results distribution: new channels of communication from radiologists to patients. AB - Radiologists face substantial challenges in the timely and appropriate communication of diagnostic test results. As with other systems in health care delivery, the radiology reporting system is not designed to be optimally safe, timely, and patient centered. To improve the quality of care, there must be a new commitment to organizing services around patients' needs, including the reporting of diagnostic test results directly from radiologists to patients. The rationale for this change is that if both referring physicians and patients are given imaging examination results from radiologists immediately after their interpretation, it would be less likely that important diagnostic information would be lost or overlooked. The results would be provided to all stakeholders in a more timely fashion, the potential for important information to "fall through the cracks" would be diminished, and safety would be improved. Providing these results to patients directly online would also allow radiologists to facilitate increased patient satisfaction and patient-centered care by treating patients as "co-customers" and equal partners with referring physicians with regard to access to information and shared decision making. PMID- 17411788 TI - Mergers involving academic medical institutions: impact on academic radiology departments. AB - During the past 10 years, there have been a number of large health care mergers in which at least one partner has been an academic medical center. This review summarizes the definitions, attributes needed for success, and reasons for failure of mergers. It then describes the various mergers and their outcomes and discusses the impact of the mergers on the involved radiology departments. PMID- 17411789 TI - The digital curriculum vitae. AB - The curriculum vitae (CV) has been the traditional method for radiologists to illustrate their accomplishments in the field of medicine. Despite its presence in medicine as a standard, widely accepted means to describe one's professional career and its use for decades as an accomplice to most applications and interviews, there is relatively little written in the medical literature regarding the CV. Misrepresentation on medical students', residents', and fellows' applications has been reported. Using digital technology, CVs have the potential to be much more than printed words on paper and offers a solution to misrepresentation. Digital CVs may incorporate full-length articles, graphics, presentations, clinical images, and video. Common formats for digital CVs include CD-ROMs or DVD-ROMs containing articles (in Adobe Portable Document Format) and presentations (in Microsoft PowerPoint format) accompanying printed CVs, word processing documents with hyperlinks to articles and presentations either locally (on CD-ROMs or DVD-ROMs) or remotely (via the Internet), or hypertext markup language documents. Digital CVs afford the ability to provide more information that is readily accessible to those receiving and reviewing them. Articles, presentations, videos, images, and Internet links can be illustrated using standard file formats commonly available to all radiologists. They can be easily updated and distributed on an inexpensive media, such as a CD-ROM or DVD-ROM. With the availability of electronic articles, presentations, and information via the Internet, traditional paper CVs may soon be superseded by their electronic successors. PMID- 17411790 TI - What is normal? A clinically useful reference collection of pediatric radiology cases created within a PACS. PMID- 17411791 TI - Doctor, would you comment on this lawsuit, please? PMID- 17411792 TI - Resident involvement in the ACR early involvement is healthy self-interest. PMID- 17411793 TI - You need a prenuptial agreement in your PACS contract: here is why. PMID- 17411794 TI - Linking accreditation and certification in medical physics. PMID- 17411795 TI - HIPAA regulations. PMID- 17411797 TI - Cardiac imaging is in your future. PMID- 17411798 TI - Access to the best possible health care is a "moral value". PMID- 17411799 TI - DEFCON one! Imaging under attack. PMID- 17411800 TI - Tear down the ivory curtain. PMID- 17411801 TI - Four more years: the White House health care agenda for 2005 to 2009. PMID- 17411802 TI - Human subjects investigation: timeless lessons of Nuremberg and Tuskegee. PMID- 17411803 TI - Medicine and business: clash of cultures or a great opportunity for the 21st century? PMID- 17411804 TI - First principles for financial modeling in medicine. AB - This article provides an overview of some of the core issues in business for physicians and is intended as an introduction. No prior understanding of business processes, finance, or accounting will be assumed or expected. The impetus for this work is the changing nature of medical practice in the United States in the past 20 years. Organizational changes and financial pressures challenge those of us in medicine as never before. For the vast majority of physicians, these realms are outside the scope of our traditional training and expertise. This article will provide an introduction to understanding these issues, starting with misconceptions about the overlap between medicine and finance. We will then introduce core concepts of cash vs. revenue, risk and uncertainty, and basic financial modeling. PMID- 17411805 TI - Financial modeling in medicine: cash flow, basic metrics, the time value of money, discount rates, and internal rate of return. AB - In this article, the authors cover tools for financial modeling. Commonly used time lines and cash flow diagrams are discussed. Commonly used but limited terms such as payback and breakeven are introduced. The important topics of the time value of money and discount rates are introduced to lay the foundation for their use in modeling and in more advanced metrics such as the internal rate of return. Finally, the authors broach the more sophisticated topic of net present value. PMID- 17411806 TI - Financial accounting for radiology executives. AB - The authors review the role of financial accounting information from the perspective of a radiology executive. They begin by introducing the role of pro forma statements. They discuss the fundamental concepts of accounting, including the matching principle and accrual accounting. The authors then explore the use of financial accounting information in making investment decisions in diagnostic medical imaging. The paper focuses on critically evaluating the benefits and limitations of financial accounting for decision making in a radiology practice. PMID- 17411807 TI - Radiology applications of financial accounting. AB - A basic knowledge of financial accounting can help radiologists analyze business opportunities and examine the potential impacts of new technology or predict the adverse consequences of new competitors entering their service area. The income statement, balance sheet, and cash flow statement are the three basic financial statements that document the current financial position of the radiology practice and allow managers to monitor the ongoing financial operations of the enterprise. Pro forma, or hypothetical, financial statements can be generated to predict the financial impact of specific business decisions or investments on the profitability of the practice. Sensitivity analysis, or what-if scenarios, can be performed to determine the potential impact of changing key revenue, investment, operating cost or financial assumptions. By viewing radiology as both a profession and a business, radiologists can optimize their use of scarce economic resources and maximize the return on their financial investments. PMID- 17411808 TI - Finance for practicing radiologists. AB - This article reviews basic finance for radiologists. Using the example of a hypothetical outpatient computed tomography center, readers are introduced to the concept of net present value. This concept refers to the current real value of anticipated income in the future, realizing that revenue in the future has less value than it does today. Positive net present value projects add wealth to a practice and should be pursued. The article details how costs and revenues for a hypothetical outpatient computed tomography center are determined and elucidates the difference between fixed costs and variable costs. The article provides readers with the steps used to calculate the break-even volume for an outpatient computed tomography center given situation-specific assumptions regarding staff, equipment lease rates, rent, and third-party payer mix. PMID- 17411809 TI - Managerial accounting applications in radiology. AB - We review the core issues in managerial accounting for radiologists. We introduce the topic and then explore its application to diagnostic imaging. We define key terms such as fixed cost, variable cost, marginal cost, and marginal revenue and discuss their role in understanding the operational and financial implications for a radiology facility by using a cost-volume-profit model. Our work places particular emphasis on the role of managerial accounting in understanding service costs, as well as how it assists executive decision making. PMID- 17411810 TI - Turf wars in radiology: emergency department ultrasound and radiography. AB - In recent years, emergency medicine physicians have made strenuous efforts to take over responsibility for interpreting and billing for ultrasound and conventional x-ray studies performed in emergency departments. This is a battle that radiologists have largely won. The authors explore some of the history of this controversy and the reasons why radiologists still control emergency department imaging. PMID- 17411811 TI - Do emergency medicine physicians perform ultrasound and conventional radiography in the emergency department? Recent trends from 1993 to 2001. AB - PURPOSE: To examine trends in emergency medicine physicians' participation in diagnostic ultrasound and conventional radiography from 1993 to 2001. METHODS AND MATERIALS: The nationwide Medicare Part B Physician/Supplier Procedure Summary Master Files for 1993 and 1996 through 2001 were examined to determine the total number of ultrasound and conventional radiography examinations performed in emergency departments, except for ophthalmic ultrasound and supervision or interpretation claims. Ultrasound examinations were categorized as general, vascular, breast, echocardiography, and obstetrical. Conventional x-ray examinations were categorized as chest, skeletal, abdomen, and gastrointestinal fluoroscopy. The total volume and overall share of the two modalities and nine aforementioned categories were calculated for each year for radiologists, emergency medicine physicians, other nonradiologists, and multispecialty groups using the Medicare provider and location-of-service codes. RESULTS: The total volume of ultrasound examinations performed in emergency departments increased from 134,533 in 1993 to 355,889 in 2001 (+164.5%). Emergency medicine physicians performed 760 (0.6%) of the examinations in 1993 and 1160 (0.3%) in 2001. Radiologists performed 87,377 (64.9%) in 1993 and 257,479 (72.3%) in 2001, other nonradiologists (primarily cardiologists performing echocardiography and vascular surgeons performing vascular ultrasound) performed 40,501 (30.1%) in 1993 and 76,649 (21.5%) in 2001, and multispecialty groups performed 5895 (4.4%) in 1993 and 20,601 (5.8%) in 2001. The total volume of conventional radiography examinations increased from 5,120,608 in 1993 to 8,054,771 (+57.3%) in 2001. Emergency medicine physicians performed 243,705 examinations (4.8%) in 1993 and 167,968 (2.1%) in 2001, radiologists performed 4,558,933 (89.0%) and 7,478,659 (92.8%), other nonradiologists performed 113,848 (2.2%) and 99,627 (1.2%), and multispecialty groups performed 204,122 (4.0%) and 308,517 (3.8%). The largest volume of examinations performed by radiologists and emergency medicine physicians in emergency departments in 2001 was in chest radiography, with radiologists performing 3,765,209 and emergency medicine physicians performing 98,851. The largest volume for other nonradiologists in emergency departments in 2001 was in echocardiography; they performed 53,943 of these examinations. CONCLUSION: Emergency medicine physicians perform a very small percentage of all ultrasound and conventional x-ray examinations performed in emergency departments, with their share decreasing over the 8-year period. The fact that the participation of emergency medicine physicians in ultrasound imaging and conventional radiography in emergency departments is limited and has been decreasing makes their claim of substantial participation highly questionable. PMID- 17411812 TI - Inspiration. PMID- 17411813 TI - Coding certification: can you afford less? PMID- 17411814 TI - Developing a business plan in radiology. PMID- 17411815 TI - Considerations for selecting a digital radiography system. PMID- 17411816 TI - ACRIN fellowship 2004: personal perspectives. PMID- 17411817 TI - Hospitals without radiologists. PMID- 17411818 TI - From research into practice. PMID- 17411819 TI - Informed and shared decision making: an alternative to the debate over unproven screening tests. PMID- 17411820 TI - Hearing the alarms: defining medical errors. PMID- 17411821 TI - Diagnostic radiology: major weapon in patient care or weapon of mass destruction? PMID- 17411822 TI - Reporting significant unexpected findings: the emergence of information technology solutions. PMID- 17411823 TI - The growing physician shortage: fact or folly? PMID- 17411824 TI - Promoting an ethical approach to unproven screening imaging tests. AB - The use of screening imaging technology such as electron beam computed tomography and computed tomographic scans for the early detection of coronary artery disease, lung cancer, and other diseases is rising, even though they have not been proven to reduce disease-specific mortality. Until randomized, controlled trials assess the efficacy of these tests as screening tests, they will remain controversial. It is unclear whether the potential benefits of these screening tests outweigh the risks. In a practice environment in which public demand and enthusiasm for screening is high, radiologists can recognize the ethical issues associated with unproven screening imaging tests; understand current national policies toward professionalism and informed and shared decision making for screening; draw on the lessons learned from the proliferation of another unproven screening test, the prostate-specific antigen blood test for prostate cancer; and work with others in the health care system to promote an ethical approach to screening imaging tests. PMID- 17411825 TI - The future of academic publishing: what is open access? AB - For more than 200 years, publishers have been charging users (i.e., subscribers) for access to scientific information to make a profit. Authors have been required to grant copyright ownership to the publisher. This system was not questioned until the Internet popularized electronic publishing. The Internet allows for rapid dissemination of information to millions of readers. Some people have seen this as an opportunity to revolutionize the system of scientific publishing and to make it one that provides free, open access to all scientific information to all persons everywhere in the world. Such systems have been launched and have instigated a wave of dialogue among proponents and opponents alike. At the center of the controversy is the issue of who will pay for the costs of publishing, because an open-access system is not free, and this threatens the backbone of the traditional publishing industry. Currently, open-access publishers charge authors a fee to have their articles published. Because of this and the uncertainty of the sustainability of the open-access system, some authors are hesitant to participate in the new system. This article reviews the events that led to the creation of open-access publishing, the arguments for and against it, and the implications of open access for the future of academic publishing. PMID- 17411826 TI - Planning for PACS: a comprehensive guide to nontechnical considerations. AB - A complete picture archiving and communication system (PACS) installation is one of the largest projects a radiology department will undertake. Although technology issues are important, they often draw focus away from many other significant issues This paper describes in detail all of these other necessary components that need to be addressed if a PACS installation is to be relatively trouble free, provides guidelines for successful PACS implementation, and details pitfalls to be avoided. PMID- 17411827 TI - Continuing medical education in radiology: a glimpse of the present and of what lies ahead. AB - The process of continuing medical education is a part of every radiologist's and radiation oncologist's professional growth, development and lifelong learning. This process aids the physician in keeping current with new techniques, procedures, and information. Continuing medical education is required for maintenance of certification. There is also a continuing medical education component and recommendations within the accreditation programs of the American College of Radiology (ACR). The ACR is accredited to sponsor educational activities and award credit hours. It sponsors educational meetings as well as enduring materials. Each activity undergoes a complex, time-consuming development process and is reviewed by the ACR's Committee on Accreditation for Continuing Medical Education. Traditional lecture based activities have not been effective with regards to improving physician performance and patient care outcomes, which are the ultimate goals of continuing medical education. The most effective educational activities are self-directed, interactive, often Web-based and available at the practice site and are timed when the information is most needed for a real life patient encounter. The ACR supports the radiology and radiation oncology community in their efforts at lifelong learning and is developing new and innovative educational programs and tools to accomplish this goal. PMID- 17411828 TI - Malpractice issues and contrast use. AB - As the cost of nonionic lower-osmolality contrast media decreases, the standard of practice for using such agents changes, and the implications of informed consent increase. Failure to use specific contrast agents appropriately or failure to respond correctly to a contrast reaction can lead to the claim of malpractice. These topics are reviewed in the context of current practice guidelines. PMID- 17411829 TI - Strategic planning in radiology. AB - As radiology continues to evolve and grow, radiologists must be concerned with preparing radiology for the future. Decisions in capital investments, mergers, outpatient diagnostic clinics, and payment and liability issues will require practicing radiologists to develop and follow up managerial, interpersonal, and learning skills that were not as necessary in the past. To become adept in the new radiology environment and be able to manage change and deal with difficult decisions, radiologists need to acquire a background in strategy. PMID- 17411830 TI - The polity of academic medicine: evidence-based democracy. AB - The authors consider the empirical data examining relationships between democratic governance and organizational success. There is overwhelming evidence that democratically run organizations excel in key parameters of success, such as business valuation, productivity, responsiveness, innovation, decision making, and worker morale and satisfaction. A review of physician surveys shows that discontent with academic administration is a major contributor to faculty turnover. Other data indicate that the basic concepts justifying autocratic governance of a department are deeply flawed and that autocratic governance is counterproductive. The authors conclude that the democratic governance of academic departments is the only model that is scientifically valid and would greatly enhance all missions of academic medicine in the 21st century. PMID- 17411831 TI - Radiologist recruitment and retention: how can we improve? AB - Although recent data suggest that the shortage of radiologists is beginning to ease, radiologists remain in high demand. Academic and private radiology practices can take specific steps both to encourage the recruitment of new radiologists and to enhance the retention of existing staff members. The authors review the history of the current workforce shortage and describe steps they have taken within their own practice that have proved successful in helping fill staff positions and prevent the significant loss of staff members over the past several years. The authors believe that these steps can be helpful to any institution, private or academic, seeking to recruit and retain radiologists. PMID- 17411832 TI - Virtues of virtual autopsy. PMID- 17411833 TI - The impact of large jury verdicts and tort reform. PMID- 17411834 TI - Buyers beware: should you purchase a modality or PACS workstation for your practice? What is the difference? PMID- 17411835 TI - How are clinical medical physics services for Medicare patients reimbursed? PMID- 17411836 TI - Self-referral: guess who is sleeping with the enemy? PMID- 17411837 TI - Re: "Why don't female medical students choose diagnostic radiology? A review of the current literature". PMID- 17411838 TI - Nothing endures but change. PMID- 17411840 TI - Why would anyone (in his or her right mind) want to do breast imaging? PMID- 17411841 TI - Vioxx, radiology, and the Food and Drug Administration. PMID- 17411842 TI - Bioinformatics, computational biology, and the National Centers for Biomedical Computing. PMID- 17411843 TI - The inappropriate use of imaging studies: a report of the 2004 Intersociety Conference. AB - The participants of the 2004 Intersociety Conference met to discuss the growing problem of self-referral. The United States spends more of its gross national product on health care than other countries, especially Japan and those in Western Europe. Imaging accounts for a large and growing portion of those costs. Despite spending so much on health care, the United States ranks relatively low in measures of national health, including such parameters as infant mortality and even life expectancy. Because the federal government must keep health care expenditures to a "sustainable growth rate," increases in use are likely to be accompanied by decreases in reimbursement per case. Thus, conference participants agreed that the real problem is inappropriate use, which may arise from (1) ignorance of what specific imaging studies are needed when, (2) high public expectations for imaging tests, (3) the fear of liability for a missed diagnosis (defensive medicine), and (4) self-referral. The Stark laws have been largely ineffective in preventing self-referral because there are many loopholes, and the laws are inconsistently enforced. Among the many potential solutions are the education of our clinical colleagues on appropriateness criteria; the education of the public on the costs of inappropriate use; tort reform; and working with third-party payers, especially the private insurance industry, to develop vigorous privileging programs, to require precertification for self-referred studies, or to establish differential payments for self-referred and non-self referred imaging. PMID- 17411844 TI - The opportunities and challenges posed by the rapid growth of diagnostic imaging. AB - As insurer representatives, the authors appreciate the benefits associated with the development and evolution of imaging. At the same time, they are concerned about the escalating costs associated with the use of these and other technologies. In 2003, Booz Allen Hamilton reported that diagnostic imaging was the most costly type of health care technology. The use of diagnostic imaging is increasing rapidly. The number of imaging procedures is projected to grow by 26% by 2008, when almost half a billion procedures will be done each year. Health plans have undertaken a variety of responses to these trends and to address concerns about maintaining the quality and affordability of care as imaging services have proliferated rapidly. The challenge for health plans, for radiologists, and for referring physicians is how to reduce the use of unnecessary or inappropriate imaging while ensuring access to clinically valuable imaging, especially in a period of rapid technological advancement and increased use. The BlueCross BlueShield Association and Blue Cross and/or Blue Shield plans are exploring ways to collaborate with the ACR and other specialty societies to promote the safe, effective, and efficient provision of imaging services. PMID- 17411845 TI - The impact of physician entrepreneurship on escalating health care costs. AB - Health care costs in this country are escalating at an alarming rate. Many economists predict this rate is unsustainable due to the long-term financial burden on our citizenry. Moreover, our health care delivery is fragmented and wasteful. United States health care is ranked last among the industrialized nations. Proponents of the U.S. system of health care extoll the virtues of our "free market." This article explores the role of physician entrepreneurship in the perversion of the marketplace of health care delivery. Medicine has become overcommercialized at the expense of patients and taxpayers. The time has come to implement legislative measures to redirect our dysfunctional health care system. This article explores the role of physician entrepreneurship in rising health care costs. Under the wrong circumstances, the invisible hand of the free market can become dysfunctional. PMID- 17411846 TI - The impact of self-referral on radiology residency training programs. AB - PURPOSE: The self-referral by nonradiologist physicians of diagnostic imaging and interventional procedures has affected radiology residency training programs. The Association of Program Directors in Radiology surveyed its members to assess the impact on specific areas of training. METHODS: The authors surveyed 186 residency program directors. RESULTS: Forty-three percent of those contacted responded. Fifty-two of 80 respondents (65%) said that in their opinion, self-referral had had an impact on residency training. Of the 52 program directors who said that their programs had been affected, 47 (90%) reported that in their opinion, self referral had a negative impact on their training programs. The areas most affected were vascular interventional radiology, obstetric ultrasound, and nuclear cardiology. CONCLUSIONS: Self-referral endangers the quality of radiology residencies. The authors discuss strategies to protect radiology residency programs. PMID- 17411847 TI - Taming the expert: standards and implications of radiologist expert witness testimony. AB - In light of the proliferation and pitfalls of expert witness testimony provided by radiologists, this article offers an overview of the standards for such testimony, as enforced by the courts, the relevant professional societies, and peer-review processes. The article also offers practical suggestions that encourage radiologist expert witnesses to be both ethical and effective in spite of the often inconsistent ambitions, expectations, and obligations of the expert's role. PMID- 17411848 TI - An overview of the ACR Committee on Ethics: from hospital contracts to expert witness testimony. AB - Professional medical ethics has challenged physicians since time immemorial. Difficult questions about whether physicians appropriately interact with patients, staff members, and their medical colleagues seldom yield ready answers. Like other professional societies, the ACR offers guidance for its members on medical ethics issues. This article discusses how the ACR Committee on Ethics serves educational and investigative and disciplinary roles. The committee has authored and contributed to articles and other resources on such topics as physician-patient communication, informed consent, and ethics training for residents. It also has received complaints that a member's conduct allegedly violated the ACR's Code of Ethics. Many recent complaints have alleged that a member failed to provide nonpartisan and accurate expert medical testimony in a legal proceeding. The committee carefully screens each complaint and has established a process for investigating and deciding whether the testimony has violated the code. PMID- 17411849 TI - Communication errors in radiology: a liability cost analysis. AB - PURPOSE: We evaluated the economic effect on radiologists involved in litigation of failures in communication of results. METHOD: We examined claims data from the Physicians Insurers Association of America 2002 report on breast cancer and identified malpractice cases in which miscommunication, rather than misdiagnosis, was the primary cause for litigation. RESULTS: The average indemnity payment for primary errors in communication by radiologists was between $228,000 and $236,000, or twice as high as when appropriate communication occurred. As a percentage of total indemnity payments to plaintiffs, such awards were 15 times higher than when communication was effective. CONCLUSIONS: Notwithstanding diagnostic accuracy, errors attributable to ineffective communication of results account for high indemnity awards. These errors can be easily resolved in clinical practice. PMID- 17411850 TI - Massachusetts radiology resident attitudes toward mammography. AB - PURPOSE: To investigate and document the attitudes toward careers in breast imaging among Massachusetts radiology residents. METHOD: Written surveys were distributed to all senior radiology residents in the state asking whether they were interested in pursuing a fellowship in breast imaging or whether they planned to interpret mammograms as part of their future practice. If they were not interested, residents were asked why. RESULTS: Of 63 residents who responded, only 1 was interested in a breast-imaging fellowship. Only 3% of radiology residents surveyed in the state agreed that they would like to spend a substantial time interpreting mammograms in the future. The most common reasons cited for not wanting to interpret mammograms in the future were "fear of lawsuits" (76%) and "not an interesting field" (49%). CONCLUSIONS: Very few Massachusetts radiology residents are interested in pursuing breast imaging in their future careers. This lack of interest in breast imaging will ultimately affect women's access to breast imaging services in the future. PMID- 17411851 TI - The current state of NIH funding of research in diagnostic radiology at U.S. medical schools. AB - The research grant awards data for 2003 published by the National Institutes of Health (NIH) were analyzed to obtain a snapshot of the current state of diagnostic radiology research in US medical schools. By considering awards to diagnostic radiology departments only, the authors show that NIH departmental rankings that combine diagnostic and therapeutic radiology departments together are deceptive for researchers interested primarily in diagnostic radiology. For each diagnostic radiology grant analyzed, the authors examined the source of funding, the primary modality involved in the research, the activity classification of the grant, and the degree of the associated principal investigator. Furthermore, the authors followed the funding for medical school radiology departments over the past several years to see if it kept up with the overall NIH budget. The data show that radiology research has exceeded the growth in academic medical research in general and highlight some areas of underfunded research. PMID- 17411852 TI - The chair as firefighter. PMID- 17411853 TI - Diagnosis please...if you want to get paid! PMID- 17411854 TI - Speech recognition system evaluation. PMID- 17411855 TI - Self-referral: what residents need to know. PMID- 17411856 TI - A look in the mirror. PMID- 17411857 TI - Pay it forward. PMID- 17411858 TI - The FDA and radiologists: momentum for change. PMID- 17411859 TI - Radiology resident applicants: are we interviewing fewer as the pool is getting larger? PMID- 17411860 TI - Veterans Affairs hospitals: prepared for duty? PMID- 17411861 TI - How HHS plans to move medical innovations forward: summary of task force report and recommendations. PMID- 17411862 TI - ACR clinical statement on noninvasive cardiac imaging. AB - Coronary artery disease and other acquired and congenital cardiac diseases are major medical and socio-economic problems. Historically, imaging has had a critical role in the diagnosis and evaluation of acquired and congenital cardiac disease. Advances in computed tomography (CT), with multidetector CT and electron beam CT technology, and magnetic resonance (MR) imaging, now make it possible to noninvasively image the coronary arteries, cardiac chambers, valves, myocardium, and pericardium and assess cardiac function, and CT and MR imaging are becoming increasingly important in the evaluation of cardiac disease. Radiologists, because of their extensive experience in CT and MR imaging, have an important role in imaging cardiac patients using these modalities. This clinical statement of the ACR discusses various technical and patient safety issues related to cardiac CT and MR imaging, and it suggests appropriate qualifications for radiologists until such time as ACR practice guidelines for the performance of cardiac CT and cardiac MR imaging are written and approved through the usual ACR process. It stresses that the interpreting physician is responsible for examining not only the cardiac structures of interest but also all the visualized noncardiac structures and must report any clinically relevant abnormalities of these adjacent structures. PMID- 17411863 TI - Outsourced teleradiology imaging services: an analysis of discordant interpretation in 124,870 cases. AB - PURPOSE: Outsourcing after-hours radiology coverage to a teleradiology coverage company has become common in recent years. However, concerns have been raised over the quality of these types of coverage and the implications on patient care. This study details the quality assurance program of a teleradiology company that provides after-hours coverage to 64 California hospitals. METHOD: The records of all examinations interpreted by 10 radiologists during 2003 were reviewed. Interpretations were compared with the final interpretations of the host practices and evaluated for timeliness. RESULTS: A total of 124,870 radiologic studies were interpreted by 10 teleradiologists during 2003. Computed tomography (CT) comprised 74% of these examinations: CT head (35%) examinations were the most commonly transmitted examinations, and CT abdomen/pelvis examinations were the second most common studies (27%). The average turnaround time was 12.2 min; 93% of the examinations were reported within 30 min, and 99% were completed within 1 hour. The overall discordant rate for individual teleradiologists ranged from 0.70% to 1.41%, with an average of 1.09%. Of the most commonly ordered examinations, CT of the abdomen/pelvis had the highest rate of discordance, at 2.1%. CONCLUSIONS: Outsourcing to a teleradiology program with an active quality assurance program can be safe. An active quality-assurance program should be an integral component of any teleradiology program. Constant feedback improves the performance of the radiologists. PMID- 17411864 TI - Technology Evaluation Center assessment synopsis: use of magnetic resonance imaging to avoid a biopsy in women with suspicious primary breast lesions. AB - Imaging technologies (e.g., magnetic resonance imaging [MRI]) have been proposed as noninvasive alternatives to surgical biopsy techniques in the follow-up of women with suspicious primary breast lesions. This article is a synopsis of a systematic review by the Blue Cross and Blue Shield Association Technology Evaluation Center. According to the best available evidence, the sensitivity and specificity of MRI ranged from 91% to 99% and from 31% to 91%, respectively. Predictive values were calculated by using three likely scenarios of diagnostic performance and disease prevalence. The potential harm of failing to diagnose or delaying diagnosis of breast cancer in patients with false-negative MRI results is of significant concern. We conclude that the potential benefit of sparing some patients from unnecessary biopsy by using noninvasive imaging does not outweigh the potential harm of a missed or delayed diagnosis of breast cancer. PMID- 17411865 TI - Effect of referring physician specialty and practice type on referral for image guided breast biopsy. AB - PURPOSE: To determine whether referring physician specialty and practice type affect the likelihood of referral for image-guided breast biopsy. METHOD: We reviewed the records of every patient who received a Breast Imaging Reporting and Data System (BI-RADS) category 4 or 5 examination (mammography, ultrasound, or both) performed during the year 2000 at both our community- and hospital-based practices. We recorded the referring physician specialty and office location (academic medical center vs. community), breast-imaging facility location (hospital vs. community), patient age, patient insurance status, BI-RADS category, and palpability of the lesion. Multiple logistic regression analysis was performed to examine the relationship of these factors to the biopsy type (surgical vs. image-guided). RESULTS: Of the 831 patients with a BI-RADS Category 4 or 5 report, 734 underwent follow-up surgical or image-guided needle biopsy. Multiple logistic regression analysis demonstrated that referral by a nonsurgeon or by a physician at the academic medical center was associated with a higher likelihood of the patient's being referred for image-guided biopsy. Patients referred by surgeons and community physicians were more likely to undergo surgical biopsy. BI-RADS Category 5 examinations and palpable lesions were also associated with a higher probability of undergoing surgical biopsy. Patient age, insurance status, and breast-imaging facility location were not related to biopsy type. CONCLUSIONS: In our practice, patients referred for breast imaging by nonsurgeons and academic physicians were more likely to be referred for image guided biopsy, whereas patients referred by surgeons and community physicians were more likely to undergo surgical biopsy. PMID- 17411866 TI - ACR MRI accreditation: yesterday, today, and tomorrow. AB - The ACR's Magnetic Resonance Imaging (MRI) Accreditation Program has generated controversy and debate since the Committee on MRI Accreditation began the development of the program in the early 1990s. This article discusses the motivation and development process for the program. It also presents outcomes from the MRI Accreditation Program, including passing and failing statistics by field strength and body part. The on-site survey process and outcomes are presented, and a case study is described. PMID- 17411867 TI - Medical devices and the US Food and Drug Administration: regulating the tools of radiology. AB - The radiology community has an ever-expanding array of technologies to use in the care of patients. Regulated by the US Food and Drug Administration, these technologies often raise complex regulatory and legal questions in everyday practice that can be daunting for practicing radiologists. This article reviews the federal medical device regulatory framework pertinent to the practice of radiology, with the aims of highlighting the potential impact of federal regulation on everyday practice and minimizing misunderstandings about enforcement exposure. PMID- 17411868 TI - Costs, charges, and revenues for hospital diagnostic imaging procedures: differences by modality and hospital characteristics. AB - PURPOSE: This study examined financial data reported by Florida hospitals concerning costs, charges, and revenues related to imaging services. METHODS: Financial reports to the Florida Hospital Uniform Reporting System by all licensed acute care facilities for fiscal year 2002 were used to calculate four financial indices on a per procedure basis. These included charge, net revenue, operating expense (variable cost), and contribution margin. Analysis, stratified by cost center (imaging modality), tested the effects of bed size, ownership, teaching status, and urban or rural status on the four indices. RESULTS: The mean operating expense and charge per procedure were as follows: computed tomography (CT): $51 and $1565; x-ray and ultrasound: $55 and $410; nuclear medicine (NM): $135 and $1138; and magnetic resonance imaging (MRI): $165 and $2048. With all four modalities, for-profit hospitals had higher charges than not-for-profit and public facilities. Excepting NM, however, the difference by ownership disappeared when considering net revenue. Operating expense did not differ by ownership type or bed size. CONCLUSIONS: Operating expense (variable cost) per procedure is considerably lower for CT than for MRI. Consequently, when diagnostically equivalent, CT is preferable to MRI in terms of costs for hospitals. If the cost structure of nonhospital imaging is at all similar to hospitals, the profit potential for performing CT and MRI seems to be substantial, which has relevance to the issue of imaging self-referral. PMID- 17411869 TI - Today's labor market: recruiting radiologists in a time of shortages. AB - Recent years have brought enormous changes to the specialty of diagnostic imaging and in particular to the radiology labor market. The impact of subspecialties, the increased use of advanced imaging, and lifestyle issues driving career related and job-related decision making by young radiologists have combined to bring about a new world of recruitment. Success in attracting, hiring, and retaining desired candidates requires acceptance of today's radiology labor market conditions. Knowledge of exactly what conditions today's entry-level candidates are seeking is key. In addition, innovation in the recruitment and development of a desirable workplace can be extremely helpful. Although these challenges may be difficult for some radiology employers to accept, it may help to know that radiology is not alone. For a wide variety of reasons, some similar, some unique to particular specialties, most medical employers today must work hard and make compromises to hire the physicians they seek. PMID- 17411870 TI - Business models for academic medical center cyclotron operations. AB - A cyclotron facility may provide a significant strategic advantage for an academic medical center that desires to build a strong research program in nuclear medicine. Such a facility may provide an advantage in obtaining support from the National Institutes of Health. A nuclear medicine research program often requires the production of short-lived radioisotopes for clinical patients. Combining the research program with a commercial production and distribution program can increase the synergies and efficiencies of an organization. This article describes various business models that combine research, clinical, and commercial operations to align an academic medical center's cyclotron program operation to its goals and resources. By coordinating these three functions, an academic medical center may be able to support extensive research capabilities that would otherwise be unattainable. PMID- 17411871 TI - The anatomy of an outstanding CME meeting. AB - Organizing a continuing medical education (CME) meeting should not be a casual undertaking. It requires experience, staff support, and adequate funding. The sponsoring entity should have a thorough grounding in the Accreditation Council for Continuing Medical Education mind-set, and the course should be within the mission of the organization. Needs of the potential attendees should be assessed and objectives developed to meet those needs. An evaluation process should be in place to ensure that the needs were addressed and met. Ideally, a postcourse evaluation should assess the impact of the meeting on the behavior and practice patterns of the registrants. Excellent teachers should be selected as faculty members. These educators should be given advance information about the characteristics of the attendees, their needs, and their expectations. Topics should be chosen for their relevance to the registrants, as opposed to the convenience of the faculty members. This paper deals with these issues and several related topics. It is based on the author's experience and supplemented by relevant peer-reviewed scientific literature. The CME "market" has a "caveat emptor" approach. PMID- 17411872 TI - Effective medical documentation. PMID- 17411873 TI - Display monitors for digital medical imaging. PMID- 17411874 TI - The noise of MRI. PMID- 17411875 TI - The ACGME Residency Review Committee for Diagnostic Radiology: a resident's perspective. PMID- 17411876 TI - Chopped liver again! PMID- 17411877 TI - It's about the patients. PMID- 17411879 TI - Maintaining and acquiring brachytherapy skills. PMID- 17411880 TI - The quickening debate over Medicaid. PMID- 17411881 TI - The potential power of reference image databases and oncologic imaging. PMID- 17411882 TI - Concurrent PET/CT with an integrated imaging system: intersociety dialogue from the Joint Working Group of the American College of Radiology, the Society of Nuclear Medicine, and the Society of Computed Body Tomography and Magnetic Resonance. AB - Rapid advances in imaging technology are a challenge for health care professionals, who must determine how best to use these technologies to optimize patient care and outcomes. Hybrid imaging instrumentation, combining 2 or more new or existing technologies, each with its own separate history of clinical evolution, such as PET and CT, may be especially challenging. CT and PET provide complementary anatomic information and molecular information, respectively, with PET giving specificity to anatomic findings and CT offering precise localization of metabolic activity. Historically, the acquisition and interpretation of the 2 image sets have been performed separately and very often at different times and locales. Recently, integrated PET/CT systems have become available; these systems provide PET and CT images that are acquired nearly simultaneously and are capable of producing superimposed, coregistered images, greatly facilitating interpretation. As the implementation of this integrated technology has become more widespread in the setting of oncologic imaging, questions and concerns regarding equipment specifications, image acquisition protocols, supervision, interpretation, professional qualifications, and safety have arisen. This article summarizes the discussions and observations surrounding these issues by a collaborative working group consisting of representatives from the American College of Radiology, the Society of Nuclear Medicine, and the Society of Computed Body Tomography and Magnetic Resonance. PMID- 17411883 TI - The ACR's Mammography Accreditation Program: ten years of experience since MQSA. AB - The ACR's Mammography Accreditation Program has been helping facilities improve the quality of mammography through peer review and professional feedback since 1987. Initially conceived as a voluntary program, accreditation became mandatory when the Mammography Quality Standards Act (MQSA) of 1992 required all U.S. mammography facilities to become accredited and certified by October 1, 1994. Currently, the ACR is the largest of four accrediting bodies approved by the U.S. Food and Drug Administration, accrediting 12,729 units at 8325 facilities by October 1, 2004. Between 1987 and 1991, 70% of the mammography units applying for accreditation with the ACR passed on their first attempts. In 2003, 88.3% of the units passed on their first attempts, indicating a marked improvement in the quality of mammography in the United States since MQSA went into effect 10 years ago. PMID- 17411884 TI - The current medical liability insurance crisis: detailed findings from two ACR surveys in 2003 and 2004. AB - PURPOSE: To describe radiologists' experiences and concerns with respect to the professional liability insurance crisis. METHODS: Data were obtained from two sources: the nationally representative 2003 ACR Survey of Radiologists and the specially undertaken 2004 ACR Malpractice Survey, a two-phase e-mail survey of ACR members. The response rate for the 2003 survey was 63%. For the two phases of the 2004 survey, response rates were low, 9% and 29%, respectively, meaning that responses probably were disproportionately from radiologists especially concerned about malpractice issues. Responses from the 2003 survey were weighted to make data representative of all radiologists in the United States. Data from the 2004 e-mail survey are not representative of all radiologists or of all ACR members. RESULTS: On the 2003 survey, 43% of radiologists said that they were less satisfied with their profession than 5 years earlier, and 70% of these cited the medicolegal climate as a major reason. This was the most commonly cited major reason for decreased satisfaction. The medicolegal climate was somewhat less important to breast imaging specialists than to other radiologists. On the 2004 survey, 65% of radiologists reported malpractice premium increases of 25% or more in the past 3 years, and 14% reported that they had stopped performing some types of studies, predominantly mammography, because of malpractice concerns. Nationally, median premiums were $20,000 annually, but the median was $30,000 or more in five states, including four very populous states. CONCLUSIONS: Malpractice liability and rising premiums are a major concern of radiologists and have led some radiologists to cease interpreting mammograms. PMID- 17411885 TI - Turf wars in radiology: the past, present, and future importance of training standards in imaging. AB - Training standards for radiologists and other physicians who wish to perform and interpret imaging have been developed by many medical organizations. There is considerable overlap, and some are contradictory. In this article, the authors review some of the existing standards and proposals for new ones and discuss how they might affect the credentialing of physicians in the near future. PMID- 17411886 TI - Physician-entrepreneurship: a user's manual, part 1: critical questions for early stage medical ventures. AB - The medical field is one of the most exciting in America for entrepreneurs. Radiologists and other medical practitioners are in a unique position to take advantage of opportunities to invent and create innovative products and services. Physicians have led many of the most important revolutionary and evolutionary changes that have occurred in the field in the past 100 years. However, many radiologists find the process of entrepreneurship a daunting one and are often deterred from acting on their ideas. This article addresses the key issues and challenges that entrepreneurs face when embarking on new enterprises. Although most of these issues face all entrepreneurs at this stage, this article focuses on medical innovation and its particular challenges. This article concentrates on the kinds of questions that physician-entrepreneurs need to ask before jumping in, the types of common pitfalls that await those who don't address these issues, and some answers to seek if you are to beat the odds and succeed. PMID- 17411887 TI - Quality assurance in radiation oncology. AB - Assuring quality in the practice of radiation oncology has become increasingly important because of the greater complexity of treatments and the advancing knowledge of the diseases that are treated. Increased public attention to errors in medicine, the increasing use of new imaging modalities to plan therapy, the availability of new therapeutic techniques such as intensity-modulated radiation therapy, and greater dependence on information technology have increased the need for intense quality assurance (QA) in the specialty. The process of QA and the resources available to radiation oncology staff members are described. PMID- 17411888 TI - Diagnostic Radiology Residency Review Committee program citations and how to avoid them. AB - The Diagnostic Radiology Residency Review Committee gives citations when a program is out of compliance with the residency program requirements of the Accreditation Council for Graduate Medical Education. Several common and some serious citations can be avoided with proactive planning by residency program directors and chairs to be in compliance. These citations include a low American Board of Radiology pass rate, a low faculty/resident ratio, a lack of timely resident evaluations, a lack of written annual evaluations of faculty members, a lack of evaluation of program curriculum and rotation objectives, inadequate faculty teaching and supervision, inadequate faculty scholarly activity, and night floats that may interfere with education. PMID- 17411889 TI - New ethical code reflects expectations for industry behavior. AB - Sporadic misbehavior, suspect sales and marketing practices, and the perceived deep pockets of the health care industry have put corporations and physicians alike at risk for investigation for fraud and abuse misconduct. The Advanced Medical Technology Association's (AdvaMed) Code of Ethics on Interactions with Healthcare Professionals addresses interactions between the technology industry and physicians, recommending appropriate behavior for partners engaged in developing, testing, learning, and applying often complex technical innovations. As this discussion shows, AdvaMed's industry code underscores uniformity between the drug and technology industries where similarities exist yet sets a distinct course where the needs of the two industries diverge. Health care professionals must be keenly aware of the differences and similarities of the overlapping codes of conducts. Provisions for allowable financial support for third-party conferences, sales and promotional meetings, industry-sponsored educational and training meetings, consulting arrangements, gifts to physicians, reimbursement for technical information, and charitable donations are all examined within the AdvaMed code of ethics and compared against codes and compliance guidance adopted by the American Medical Association; the Pharmaceutical Research and Manufacturers of America; and the US Department of Health and Human Services; Office of Inspector General. PMID- 17411890 TI - Varicose vein treatment: considerations for a radiology group practice. AB - Varicose vein management presents an exciting opportunity for the expansion of the interventional components of many practices. Although vein practice development has been addressed by other specialists, concerns unique to radiologists have only lately become important. Radiology groups are well suited to support the addition of this service line, and it may serve as a showcase for further clinical growth. Success depends on appropriate business planning, clinical and administrative infrastructure, and intelligent marketing. This article describes requisites for the successful development of a phlebology practice by a radiology group. PMID- 17411891 TI - When expert coding advice isn't. PMID- 17411892 TI - The worth of a leader. PMID- 17411893 TI - Changes in equipment acceptance testing. PMID- 17411894 TI - Twenty-four-hour attending physician coverage and its impact on resident training. PMID- 17411895 TI - Sun Valley Group manifesto. PMID- 17411896 TI - The public face of radiology. PMID- 17411897 TI - Physicians are only human... PMID- 17411898 TI - Case for the physician executive. PMID- 17411899 TI - Pay for performance: at last or alas? PMID- 17411900 TI - National Center for Research Resources: catalyst for discovery. PMID- 17411901 TI - Changes in procedure volume and service provider distribution among radiologists and nonradiologists in dual-energy x-ray absorptiometry between 1996 and 2002. AB - PURPOSE: To determine changes in the number of dual-energy x-ray absorptiometry (DEXA) procedures performed in the United States, as well as changes in the relative number of procedures performed by radiologists and nonradiologists. METHODS: The two major Current Procedural Terminology, Fourth Edition, codes for DEXA scans were selected from the national Medicare Part B databases from 1996 through 2002 (the most recent). The total number of scans done on Medicare patients, as well as the percentages performed by radiologists and nonradiologists, were tabulated. RESULTS: In 1996, a total of 501,505 DEXA scans were performed among all Medicare patients. Of these, 30% were performed by radiologists. In 2002, there were 2,195,548 DEXA scans performed among all Medicare patients, of which radiologists performed 45%. CONCLUSION: Between 1996 and 2002, the number of DEXA scans performed on all Medicare patients increased by more than fourfold. In the same population, the percentage of scans performed by radiologists increased from less than one-third to almost one-half. Consequently, radiologists are steadily gaining market share of this procedure in the Medicare population. PMID- 17411902 TI - Get paid for what you do: dictation patterns and impact on billing accuracy. AB - No one would knowingly invest in a business whose principal executives knew little or nothing about the key components determining reimbursement for the services provided. Superimpose on that lack of knowledge a regulatory environment in that business sector that places owners and key employees at risk for accusations of fraud and abuse as well as in jeopardy of large fines and potential exclusion from the marketplace for the largest consumer of a company's product if billing is done incorrectly. Yet this is exactly the case in many radiology practices today. A significant number of radiologists who provide excellent quality medical care produce dictated reports that demonstrate complete ignorance of the parameters used by their billing personnel to generate accurately coded claims, thus losing significant legitimate clinical practice revenues while placing themselves and their practices in jeopardy. This article does not outline ways to game the system or inappropriately augment practice revenues. Rather, it describes many of the basic elements needed in the dictated reports produced by radiologists in their daily work, calculates examples of the financial impact of medically correct but poorly documented reports, and provides dictation guidelines for radiology residents and radiologists in practice that, if adopted, should ensure that you get paid properly for what you do. PMID- 17411903 TI - Hippocratic Economics (the Physician's Dilemma). AB - The nature of the medical profession puts physicians in an unusual position. Patients seek out physicians' help because of their expertise in dealing with illnesses, possibly even life-threatening ones. The asymmetry of knowledge in this relationship, the expert physician and the inexpert patient, creates an ethical dilemma for physicians regarding the delivery of care. Physicians determine how much care to offer while receiving remuneration for this care. Here, acting as patients' agents, physicians have immense discretionary power not only with patients' health but also with their pocketbooks. Known as the principal-agency problem, this type of relationship is part and parcel of what business scholars refer to as moral hazard. This article explains the problem of moral hazard and how it affects radiologists and places it in the context of professional and ethical behavior. Its causes and relationship to human nature are explored. The consequences of falling prey to moral hazard in the practice of radiology are discussed. PMID- 17411904 TI - Stereotactic body radiation therapy. AB - Stereotactic body radiation therapy constitutes an emerging therapeutic paradigm. These treatments are unique relative to the large body of experience with conventional fractionated radiotherapy. On the basis of the treatment principles of intracranial stereotactic radiation combined with enhancements associated with immobilization and imaging, the role of extracranial treatment continues to evolve. However, most clinical reports on extracranial treatments suffer from short or incomplete follow-up, making final assessments of benefit and toxicity, particularly late toxicity, problematic. These techniques are centered on a very basic understanding of the use of ionizing radiation for the treatment of cancer. Nonetheless, they do require a unique and special understanding of radiobiologic and physics principles. It is hoped that using high-dose, single-fraction treatment or a few fractions of treatment, the therapeutic ratio is improved, thus potentially changing the way some cancers are treated. Ideally, all patients receiving such treatments would be enrolled in formal protocols. As data accrue and understanding of these techniques improve, it will be possible to better define the indications for stereotactic body radiation therapy. At that time, appropriate applications can be submitted for permanent billing codes that will describe a process of care that embraces this technology without vendor favoritism. This review summarizes the state of stereotactic body radiation in 2005. PMID- 17411905 TI - Challenges facing radiology educators. AB - This article summarizes the current challenges faced by radiology educators. These are discussed under the headings of time available for education, financing education, what and how to teach, student evaluation, support infrastructure, and challenges to leadership. PMID- 17411907 TI - Pennsylvania's Patient Safety Authority. AB - The objective of this article is to improve patient safety by examining the Pennsylvania Patient Safety Authority, which monitors a statewide reporting system for medical errors and issues statewide advisories suggesting methods for reducing the types of medical errors identified on the basis of the analysis of data reported. First, the article explores the foundation for the Patient Safety Authority and discusses how it reflects many of the recommendations by the Institute of Medicine in To Err Is Human. Second, the article examines the legislation that established the Patient Safety Authority. Third, it examines the development and benefits of the Patient Safety Authority worth emulating. Fourth, the article identifies some of the weaknesses of the Patient Safety Authority. Last, the article concludes that the Pennsylvania Patient Safety Authority exemplifies a major step forward on a long and difficult path toward patient safety. PMID- 17411908 TI - Neuroradiology fellowship match: year 4 applicants' perceptions. AB - BACKGROUND AND PURPOSE: The selection of neuroradiology fellows who will start on July 1, 2005, marked the fourth year in which program directors in neuroradiology used the National Resident Matching Program (NRMP) as the selection modality. The aim of this study was to investigate fellowship candidates' views regarding the match process. METHODS: A Web survey was distributed electronically to all candidates applying for neuroradiology fellowship positions offered for the July 1, 2005, start date, with questions about the match. One hundred thirteen neuroradiology fellowship candidates responded to the survey, representing 73.9% of the surveyed fellowship pool. Candidates were also allowed to enter their comments regarding the match process via the Web survey. RESULTS: Of the 113 neuroradiology fellowship candidates responding, 75.7% felt that the NRMP system was fair, 86.0% were satisfied with the outcome, 88.8% had matched with one of their top three choices, 54.5% recommended continuing the neuroradiology fellowship match, and 15.5% stated that they experienced violations of the match rules. The main perceived advantages of the match from the candidates' perspectives were the ability to see many programs and explore multiple options, the perception of fairness, and the efficiency of using the match program. The main disadvantages were the cost of applications and traveling, the number of days required to interview, and the perception that the match program favors internal candidates. CONCLUSION: The majority of fellowship candidates felt that the match system for selection is a fair process and were satisfied with the outcome. However, the perceived disadvantages produced tepid (54.5%) support for the continuation of the match. Program directors in neuroradiology and other radiologic subspecialties should consider the data provided in this article in deciding the future selection process. PMID- 17411909 TI - Development of an interactive model for teaching emergency pediatric radiography: preliminary report. AB - PURPOSE: The authors describe the development of an interactive digital teaching module designed to help prepare residents to diagnose emergency pediatric radiology cases. METHODS: Cases were identified using the authors' own dictation search software. Cases were selected that depicted common conditions that had been misdiagnosed by the radiology resident who had first interpreted them clinically. Normal cases involving similar anatomic regions were also collected. Images from these cases were captured from the picture archiving and communication system. "Hotspots" were superimposed on abnormalities to highlight them. Example dictations were captured. A simple user interface was developed for the teaching module, and a database was built to log all user responses. RESULTS: The system has been well received at the authors' institution. It has been incorporated into the standard orientation for all incoming residents. Residents' diagnostic performance has subjectively improved since the module was launched. CONCLUSIONS: The module is interactive, easy to use, and subjectively improves incoming residents' diagnostic accuracy. A prospective, controlled study assessing its impact on short-term and long-term diagnostic performance is under way. PMID- 17411910 TI - The appeal. PMID- 17411911 TI - Role and status of information technology solutions in radiology reporting. PMID- 17411912 TI - The headache that has HIPAA written all over it! PMID- 17411913 TI - Radiation dose and evidence-based regulation. PMID- 17411914 TI - George E. Pfahler. PMID- 17411915 TI - With skill and safety. PMID- 17411916 TI - The new emphasis on quality and safety. PMID- 17411917 TI - Getting at the facts on imaging utilization growth. PMID- 17411918 TI - Interventional radiology: transformation into a clinical discipline. PMID- 17411919 TI - Still human after all these years. PMID- 17411920 TI - Recent trends in utilization of cardiovascular imaging: how important are they for radiology? AB - PURPOSE: To determine (a) the role cardiovascular imaging (CVI) plays within the spectrum of noninvasive diagnostic imaging (NDI), (b) the role radiologists play in CVI, and (c) recent trends in CVI workload among radiologists and cardiologists. METHODS AND MATERIALS: The national Medicare Part B databases for the years 1993 and 2002 were used to identify all CPT-4 codes pertaining to NDI. Those codes relating specifically to noninvasive CVI were grouped together in seven categories. The 2002 Medicare total professional component relative value units (RVUs) were then used to determine RVU rates per 1,000 Medicare beneficiaries as a measure of physician work for each code. The percentage of all NDI RVUs accruing to CVI in 2002 were calculated. Also calculated were the 9-year changes in RVU rates for each of the seven CVI categories, and the changes in those rates among radiologists and cardiologists. RESULTS: CVI represented 29% of all Medicare NDI work in 2002. Between 1993 and 2002, the RVU rate for all CVI rose by 75%, compared with 34% for all non-cardiovascular imaging. Over the same years, the CVI RVU rates among cardiologists grew by 110%, compared with 50% among radiologists. In 2002, cardiologists performed 448 RVUs per 1,000 in CVI, (mostly in echocardiography and cardiac nuclear imaging) whereas radiologists performed 162. The most rapid growth among the major CVI categories was in cardiovascular nuclear imaging performed by cardiologists; this grew 452% between 1993 and 2002. Radiologists predominate in vascular US and cardiovascular CT and MRI. CONCLUSION: CVI represents a large and important component of total diagnostic imaging services and is growing more rapidly than other types of imaging. Cardiologists dominate in CVI RVU rates and their utilization is growing more rapidly than that of radiologists'. However, radiologists predominate in vascular US and cardiovascular CT and MRI. SUMMARY SENTENCE: Cardiovascular imaging represents almost one-third of total noninvasive diagnostic imaging services and is growing more rapidly than other types of imaging; the authors explore the relative roles of radiologists and cardiologists in this field. PMID- 17411921 TI - Establishing a cardiac MRI program: problems, pitfalls, expectations. AB - Magnetic resonance imaging (MRI) has been used to evaluate the cardiovascular system for almost 2 decades. Although vascular applications have been robust and steadily improving for many years, the utility of MRI for clinical cardiac imaging has been limited. However, recent advances in hardware technology and pulse sequence design have led to substantial improvements in image quality, while reducing scan times to clinically reasonable durations. Pulse sequences using electrocardiographic gating and k-space segmentation have made it possible to obtain high-contrast, high-resolution images of the beating heart within single breath-holds. These images in turn have provided unprecedented visualization of myocardial morphology and function. Because of these developments, cardiac MRI (CMR) has made rapid and dramatic inroads into the clinical arena. Currently, the primary limitations to routine clinical application are hardware availability, clinical acceptance, politics, examination cost, and not least of all physician education. As these limitations are overcome or made more manageable, the clinical use of CMR will grow, potentially without bound. Combined with steady hardware development and an ever-growing armamentarium of pulse sequences, MRI may ultimately become the modality of choice for cardiac imaging. Because cardiac imaging is relatively unknown territory for most radiologists and because the high-end equipment has until now been sparsely available, CMR has been largely limited to major medical centers. However, if sufficient interest is present and resources are appropriately allocated, CMR can be successfully implemented in community imaging practices. PMID- 17411922 TI - Turf wars in radiology: should it be radiologists or cardiologists who do cardiac imaging? AB - In recent years, the cardiac capabilities of computed tomography, magnetic resonance imaging, and to a lesser extent positron emission tomography have rapidly advanced. This has led to contention between radiologists and cardiologists over who should perform and interpret these studies. The authors present the arguments favoring both sides and discuss strategies radiologists should pursue. Although the arguments favoring radiologists are more compelling, this may be an instance in which both sides should work together. PMID- 17411923 TI - Image-based brachytherapy: a forum for collaboration between radiation oncologists and diagnostic radiologists. AB - There has been increased interest in implementing image-guided brachytherapy to better define the structures of interest and assess the radiation dose distribution in tumors and surrounding normal tissues. This is particularly helpful in the treatment of pelvic malignancies such as cervix cancer and prostate cancer, in which the tumor lies in close relationship to the bladder and rectosigmoid. This provides a forum for the collaboration of diagnostic radiologists and radiation oncologists. PMID- 17411924 TI - Digital mammography: current capabilities and obstacles. AB - Digital mammography represents an exciting new technology for breast imaging and possibly breast screening. The decoupling of functional components in digital mammography translates into potential operational efficiencies compared with screen-film mammography (SFM). Digital mammography is a platform for advanced applications not possible with traditional SFM. However, for digital mammography to replace SFM in daily clinical practice, operational and clinical hurdles will have to be overcome. PMID- 17411925 TI - Illustrating the root-cause-analysis process: creation of a safety net with a semiautomated process for the notification of critical findings in diagnostic imaging. AB - The ACR has set a standard for the communication of critical findings on imaging examinations. Despite this standard, for a variety of reasons, it remains possible that appropriate follow-up is not initiated. The authors review the theory and application of root-cause analysis to such a failure of communication within their institution, including the development and implementation of a semiautomated notification system for critical unexpected findings on imaging examinations. PMID- 17411926 TI - Pay for performance: pay more or pay less? AB - Several recent publications in the radiology literature have reported on the growing phenomenon of pay for performance. This potent new business model seeks to reward health care providers with financial incentives for improvements in the performance and delivery of medical services. This paper briefly reviews some of the obstacles to the implementation of this strategy in the practice of radiology. Radiologists are encouraged to participate actively with payers in discussions about improving quality care but should carefully consider the potential contractual implications associated with these initiatives. PMID- 17411927 TI - Analysis and practical use: the Abt Study of Medical Physicist Work Values for Radiation Oncology Physics Services--round II. AB - PURPOSE: The initial Abt Study of Medical Physicist Work Values for Radiation Oncology Physics Services was published in October 1995. That study measured qualified medical physicist (QMP) work associated only with routine radiation oncology procedures. In the intervening years, medical physics practice has changed dramatically. Three-dimensional treatment planning, once considered a special procedure, is the standard of care for many patient presentations. Prostate seed brachytherapy, stereotactic procedures, and intensity-modulated radiation therapy now constitute a large portion of the time medical physicists devote to clinical duties. Special procedures now dominate radiation oncology, leading to the request for an updated work and staffing study for qualified medical physicists. METHODS: The updated Abt Study of Medical Physicist Work Values for Radiation Oncology Physics Services: Round II was published in June 2003. Round II measures and reports QMP work associated with both routine and most contemporary special procedures. Additionally, staffing patterns are reported for a variety of practice settings. RESULTS: A work model is created to allow medical physicists to defend QMP work on the basis of both routine and special procedures service mix. The work model can be used to develop a cost justification report for setting charges for radiation oncology physics services. The work and cost justification models may in turn be used to defend medical physicist staffing and compensation. CONCLUSION: The updated Abt study empowers medical physicists to negotiate service or employment contracts with providers on the basis of measured national QMP work force and staffing data. PMID- 17411928 TI - The National Institute of General Medical Sciences. PMID- 17411929 TI - Another unpaid second opinion. PMID- 17411930 TI - Medical students are our future. PMID- 17411931 TI - Digital mammography: coming of age. PMID- 17411932 TI - Merrill C. Sosman. PMID- 17411934 TI - Re: "Outsourced teleradiology imaging services: an analysis of discordant interpretation in 124,870 cases". PMID- 17411935 TI - The wrong fork in the road. PMID- 17411936 TI - Individual rights and the good of the specialty. PMID- 17411937 TI - Should the ordering of medical imaging examinations be reexamined? PMID- 17411938 TI - Specialty hospitals hang in the balance. PMID- 17411939 TI - Restructuring cancer clinical trials. PMID- 17411940 TI - Wireless capsule endoscopy in patients with obscure small-intestinal bleeding. AB - Wireless capsule endoscopy is proposed in the management of patients with obscure gastrointestinal (GI) bleeding, defined as persistent or recurrent GI bleeding without a cause identified on routine endoscopy. Obscure GI bleeding often originates from a small-intestinal abnormality, which presents diagnostic and therapeutic challenges because the small intestine is not easily visualized on conventional workup. This is a synopsis of a systematic review by the Blue Cross Blue Shield Association Technology Evaluation Center. Limited evidence shows that capsule endoscopy identified small-bowel lesions generally beyond the reach of push enteroscopy in 25% to 50% of patients and revealed additional diagnostic findings in 25% of patients compared with small-bowel barium radiographic studies. The authors conclude that in some cases, the information provided by capsule endoscopy in this specific patient population with obscure GI bleeding not diagnosed via conventional workup can lead to changes in management that would improve net health outcome. PMID- 17411941 TI - Comparison of recent utilization trends in radionuclide myocardial perfusion imaging among radiologists and cardiologists. AB - PURPOSE: To study recent practice patterns in radionuclide myocardial perfusion imaging (MPI) and related procedures among radiologists, cardiologists, and other physicians. METHODS AND MATERIALS: The nationwide Medicare administrative Part B claims summary databases from 1998 and 2002 were used to assess utilization rate changes in the 4 primary procedure codes for MPI, the seven codes for diagnostic cardiac catheterization with coronary angiography, and the single code for stress echocardiography. Rate changes among radiologists, cardiologists, and other physicians were determined for the total Medicare population, as well as for the 3 primary places of service at which imaging is formed: hospital inpatient facilities, hospital outpatient facilities, and private offices. Ratios of the use of the 2 supplementary codes for left ventricular (LV) wall motion and ejection fraction to that of the primary MPI codes were calculated for 2002. RESULTS: The utilization rate per 1000 Medicare beneficiaries of MPI rose among radiologists from 19.8 in 1998 to 20.1 in 2002, a 2% increase. The rate among cardiologists rose from 22.9 in 1998 to 40.7 in 2002, a 78% increase. Most of this growth occurred in cardiologists' offices, where the utilization rate increased 101% over the 4 years. In 2002, the ratios of the use of the supplementary LV wall motion and ejection fraction codes to that of the primary MPI codes were 1.73 for cardiologists and 1.46 among radiologists. Between 1998 and 2002, the utilization of diagnostic cardiac catheterization among cardiologists increased by 19%, and their utilization of stress echocardiography increased by 21%. CONCLUSION: In recent years within the Medicare population, the rate of utilization of MPI among radiologists has remained relatively stable, whereas it has risen sharply among cardiologists. The greatest growth was seen in cardiologists' private offices. This raises concerns about possible inappropriate utilization of MPI and also about the potential effect self-referral has on this utilization trend. The increased use of MPI by cardiologists did not result in reduction in their use of either cardiac catheterization with coronary angiography or stress echocardiography. PMID- 17411942 TI - Teaching with a personal digital assistant. AB - Personal digital assistants (PDAs) are powerful educational tools because of their small, mobile form factor and wide range of available applications. Ongoing technological improvements and innovations will provide compelling validation of the use of PDAs in the educational endeavors of every academic radiologist. This article describes both common and advanced features enabling PDAs to be used as effective just-in-time educational tools. PMID- 17411943 TI - The food and drug administration and molecular imaging agents: potential challenges and opportunities. AB - Molecular imaging, which combines the use of traditional imaging modalities with pharmaceutical and biologic imaging agents, holds considerable promise, particularly in light of recent advances in genomics and new applications beyond diagnosis toward the earlier detection and characterization of disease, the reliable assessment of treatment efficacy, and imaging-link therapeutic applications. Its potential notwithstanding, the current U.S. Food and Drug Administration (FDA) regulatory framework governing imaging agents sets a high bar for marketing approval, which may slow the pace at which molecular imaging becomes routinely available. Understanding the FDA's regulatory framework, the issues surrounding molecular imaging agents, and potential opportunities will allow the radiology community to more effectively collaborate with the FDA to address these regulatory barriers and ensure that the full potential of molecular imaging is realized. PMID- 17411944 TI - Should an academic radiation oncologist be allowed to "opt out" of medicare? AB - Federal law permits physicians to "opt out" of Medicare. When a radiation oncologist chooses this option, he or she may neither bill nor collect from Medicare, but may legally attempt to charge and collect what he or she considers the value of services provided to Medicare-eligible patients. Many academic faculty practice plans permit members to opt out. Even if it is permissible for a radiation oncologist to opt out of Medicare, is it appropriate? The question raises significant ethical and economic issues as one attempts to balance the good of the individual faculty member against the good of the clinical faculty as a whole. In this commentary, the authors offer the principal arguments in favor of and against permitting a faculty radiation oncologist to opt out. They conclude by recommending broad faculty oversight over such decisions. PMID- 17411945 TI - Negotiating in a managed care world. AB - Medical managed care, once thought to be a passing influence affecting large urban markets only, has proven to be a dominating factor in virtually every medical practice in the country. Discounted rates, steerage, utilization management, pay for performance, and other managed care strategies are likely to be a provider's reality for the foreseeable future. It is imperative that physicians develop negotiating skills and educate themselves about how to negotiate not only rates but also the other components discussed herein that ultimately determine the economic viability of a managed care agreement. PMID- 17411947 TI - The radiology class of 2005: postresidency plans. AB - PURPOSE: To examine the postresidency plans of the radiology residency class of 2005, including influences on and trends in the decision to pursue subspecialty fellowship training after residency. METHODS: An electronic survey composed by the ACR Resident and Fellow Section was completed by 416 senior residents (a 46.9% response rate) enrolled in allopathic radiology residency programs from across the United States, representing approximately 41.2% of the entire residency class. RESULTS: Eighty-seven percent (95% confidence interval 83.8% to 90.2%) of respondents plan to do fellowships immediately after residency, representing an increasing number, compared with published figures from the 1990s. Increasing age at the completion of residency was independently and significantly correlated with the decision not to pursue a fellowship. There has been a shift in the most popular fellowships, with an increased interest in musculoskeletal radiology fellowships and decreased interest in interventional radiology. Among residents not pursuing immediate fellowship training, the most common plans were private practice positions (48.1%), military obligations (22.2%), and nighthawk positions (9.3%). CONCLUSION: An increasing number of residents are electing to pursue subspecialty training after the completion of residency. Close monitoring of the postresidency plans of current residents may portend important trends in education, residency training, and changes in the radiology workforce. PMID- 17411948 TI - Risks associated with outside radiographs. PMID- 17411949 TI - Upgrading software in your practice: basics for success. PMID- 17411950 TI - Selecting a medical physics consultant. PMID- 17411951 TI - Harry Garland. PMID- 17411953 TI - Re: "Technology evaluation center assessment synopsis: use of magnetic resonance imaging to avoid a biopsy in women with suspicious primary breast lesions". PMID- 17411954 TI - Are residents' on-call errors worse than the alternatives? PMID- 17411956 TI - Professionalism. PMID- 17411957 TI - Brave new world. PMID- 17411958 TI - Radiology recruiting: easier said than done. PMID- 17411959 TI - Leveraging the synergy between industry and academia. PMID- 17411960 TI - Health care and public opinion. PMID- 17411961 TI - Recent developments in cancer therapy and why we need a human cancer genome project. PMID- 17411962 TI - The role of negative CT of the abdomen and pelvis in the decision to admit adults from the emergency department after blunt trauma. AB - RATIONALE AND OBJECTIVES: Computed tomography (CT) is widely used in the initial evaluation of blunt trauma patients and is associated with a high rate of negative imaging. A described benefit of negative imaging is prompt discharge. This study examined a single level 1 trauma center to determine whether adult blunt trauma patients are discharged from the emergency department (ED) after negative CT of the abdomen and pelvis (CT AP). MATERIALS AND METHODS: The authors retrospectively created a data set of adult blunt trauma patients who received CT AP in the ED from August to November 2003. Statistical analysis of admission rates on the basis of positivity or negativity on CT AP was performed to determine if the test influenced admission rates. Additional subgroup analysis was made between the patients admitted with negative CT AP and those who were discharged from the ED. RESULTS: Two thirds (316/469) had negative CT AP. Whereas 80.4% of the patients (254/316) with negative CT AP were admitted, 98.0% (148/151) with positive CT AP were admitted, a statistically significant difference in admission rate (P < .0001). The vast majority (208/254, 81.9%) of patients with negative CT AP were admitted for extra-abdominal injuries. There was no statistical difference in the characteristics of a subgroup of 45 patients who were admitted without any documented injuries from the group discharged from the ED in terms of age, gender, comorbidity, Glasgow Coma Scale score, or intoxication. CONCLUSION: Under current practice, negative CT AP after blunt trauma results in a statistically significant decrease in admissions. PMID- 17411963 TI - Medicare reimbursement for physician services: the current status of local payment policy. AB - Keeping up with the technical and academic advances in medicine of the past 2 decades has made studying the US government's physician reimbursement system a low priority for most physicians. However, in the current environment of declining physician reimbursement and increasing frequency of compliance audits by Medicare, it is important for all physicians to have a basic understanding of the Medicare payment process. A major component of the physician payment system occurs at the local level. Through local coverage determinations, state Medicare contractors make more than 90% of all Medicare coverage decisions. Federal law requires Medicare contractors to seek physician input into their coverage decision process through contractor advisory committees, and through these committees, physicians can have significant influence over the coverage decision process. Once local contractors have made their coverage decisions, the covered indications for a procedure or treatment are published for the provider community. At that point, it becomes the responsibility of physicians to know the covered indications for certain services, because contractors will deny claims for services that are not linked to covered indications. This review focuses on the basics of the local Medicare payment process, with emphasis on the development of local coverage decisions by contractors. This understanding will allow physicians to positively influence the local reimbursement process. PMID- 17411964 TI - How much do radiologists and radiation oncologists specialize? AB - PURPOSE: To present detailed information about the actual, current extent of the subspecialization of radiologists and radiation oncologists. METHODS AND MATERIALS: The authors used data from the ACR's 2003 Survey of Radiologists and 2003 Survey of Radiation Oncologists. Multiple measures were used to ascertain the overall extent of the subspecialization of radiologists and radiation oncologists. Also, the percentages of radiologists and radiation oncologists in each subspecialty are reported. Multiple regression analysis identified the probable effects on specialization of characteristics of radiologists and radiation oncologists and the practices they work in. RESULTS: Overall, 69% of posttraining, professionally active radiologists specialized at least to a small extent. Thirty-one percent of radiologists spent at least 70% of their work time in a single subspecialty. After controlling for other factors considered in the regression analysis, younger radiologists, those having taken fellowships, and those working in academic practices were more likely to be specialized; solo and locum tenens radiologists were less likely to be specialized. Forty-eight percent of radiation oncologists specialized at least to a small extent, whereas only 11% spent at least 70% of their work time in subspecialties. Controlling for other factors, radiation oncologists between 45 and 54 years of age had a low likelihood of being specialized, and the effects of practice type were the same as for radiologists. CONCLUSIONS: Depending on the measure used, between one third and two thirds of radiologists were specialized in 2003. Radiation oncologists were less specialized, but approximately half of them specialized at least to a small extent. PMID- 17411965 TI - The needs of an educator. AB - A medical educator has needs that are specific to educators and needs that are common to all medical faculty members. An educator needs time to perform educational duties; space to carry out educational activities; and money to buy time, space, and other resources. Just as important as time, space, and money are to the success of an educator is having an infrastructure that supports the educator and the educational mission. Such an infrastructure includes a system that provides educational leadership opportunities, institutional support for medical education, opportunities and funding for medical education research, students, feedback from students and peers, faculty development and mentoring, national societal support for education, and an institutional agenda that values education to the degree that it values patient care and investigative research. PMID- 17411966 TI - Ethical, legal, and social issues related to genomics and cancer research: the impending crisis. AB - Cancer research is a multibillion-dollar enterprise validated by the clinical trial process and increasingly defined by genomics. The continued success of the endeavor depends on the smooth functioning of the clinical trial system, which in turn depends on human subject participation. Yet human subject participation can exist only in an atmosphere of trust between research participants and research sponsors, and the advent of genomics has raised a multitude of ethical, legal, and social issues that threaten this trust. The authors examine 6 of these issues: (1) informed consent; (2) privacy, confidentiality, and family disclosure dilemmas; (3) property rights in genomic discoveries; (4) individual and institutional conflicts of interest; (5) insurance and employment issues; and (6) litigation under the federal False Claims Act. The authors conclude that failure to resolve these issues may lead to a sufficient impairment of trust in genomics based clinical trials on the part of potential research participants that the clinical trial system may implode for lack of willing participants, thus threatening the future of cancer research. PMID- 17411967 TI - The role of open-source software in innovation and standardization in radiology. AB - The use of open-source software (OSS), in which developers release the source code to applications they have developed, is popular in the software industry. This is done to allow others to modify and improve software (which may or may not be shared back to the community) and to allow others to learn from the software. Radiology was an early participant in this model, supporting OSS that implemented the ACR-National Electrical Manufacturers Association (now Digital Imaging and Communications in Medicine) standard for medical image communications. In radiology and in other fields, OSS has promoted innovation and the adoption of standards. Popular OSS is of high quality because access to source code allows many people to identify and resolve errors. Open-source software is analogous to the peer-review scientific process: one must be able to see and reproduce results to understand and promote what is shared. The authors emphasize that support for OSS need not threaten vendors; most vendors embrace and benefit from standards. Open-source development does not replace vendors but more clearly defines their roles, typically focusing on areas in which proprietary differentiators benefit customers and on professional services such as implementation planning and service. Continued support for OSS is essential for the success of our field. PMID- 17411968 TI - Radiology residency call: residents' perceptions of the on-call experience. AB - PURPOSE: This paper focuses on specific resident perceptions of call, including the compliance of training programs with the Accreditation Council for Graduate Medical Education's (ACGME) resident work rules and the level of comfort residents have in contacting out-of-house staff radiologists for help during on call hours. METHODS: The authors designed a Web-based survey to examine radiology residents' perceptions about call conditions with input from the 2003-2004 ACR Resident and Fellow Section Executive Council. The survey instrument was distributed to chief residents in the United States and Canada with the help of the ACR. RESULTS: Eighty-five percent of respondents felt that the amounts of work and responsibility required for call were appropriate to their levels of training. Ninety-eight percent of US respondents felt that their programs were in complete compliance with the ACGME's resident work guidelines. Eighty-nine percent were comfortable contacting the staff physicians on call when those staff physicians were out of the hospital. Respondents who felt uncomfortable calling their staff physicians were more likely to feel that the work and responsibility of call were excessive to their levels of training (chi(2) = 11.301, P = .0033). CONCLUSIONS: The majority of residents indicated that they were adequately trained for call. Most residents were on call without in-house staff coverage but felt comfortable contacting on-call staff physicians if needed. Residents who felt uncomfortable contacting on-call staff physicians were more likely to feel that their training had not prepared them for call. A high percentage of radiology resident programs are in compliance with the ACGME's resident work guidelines. PMID- 17411969 TI - The radiology class of 2005: the fellowship application process and perceptions of the NRMP fellowship match. AB - PURPOSE: To assess experiences and perceptions of the fellowship application process of the radiology residency class of 2005, including the 2005 National Resident Matching Program (NRMP) Radiology Fellowship Match. METHODS: An electronic survey composed by the ACR Resident and Fellow Section was completed by 416 senior residents (a 46.9% response rate) enrolled in allopathic radiology residency programs from across the United States. RESULTS: Residents generally agree with the timing of the current fellowship application calendar in the spring of the third year of residency. Respondents were divided over the issue of permitting internal candidates to fill positions outside of the match, although the majority felt that the process influences both applicants and programs to favor their own institutions to avoid the match process. Residents support requiring fellowship programs to declare the number of positions being filled through the match before applications are due (72.6%) or before rank lists are due (79.8%). Residents noted match rule violations, which threaten to undermine residents' confidence in the process. Residents were split over the future of the match: 23.1% favored continuing the match in its current form, 28.8% favored continuing the match with modifications, and 34.4% favored discontinuing the match entirely. CONCLUSION: Although the NRMP Fellowship Match was created to improve and standardize the fellowship application process, and although a majority of residents support continuing the match in some form, there are areas of concern, including adherence to the letter and spirit of the match rules, which must be addressed to galvanize support among residents in the future and ensure a fair process for fellowship applicants. PMID- 17411970 TI - Promoting spirited discussion. PMID- 17411971 TI - New CT codes to take to heart. PMID- 17411972 TI - Outside images on CD: a management nightmare. PMID- 17411973 TI - Charles T. Dotter. PMID- 17411974 TI - Re: commentary on "Challenges facing radiology educators". PMID- 17411975 TI - The quality promise. PMID- 17411976 TI - Dumb and dumber. PMID- 17411977 TI - The nighthawk: bird of paradise or albatross? PMID- 17411978 TI - Visibility matters. PMID- 17411979 TI - The vital signs of quality improvement organizations. PMID- 17411980 TI - 2005 ACR presidential oration: collective wisdom: lessons for the future. AB - Review of presidential orations delivered since the inception of the ACR in 1923 yields insight into problems that have faced radiology over the years and the aggressive way they have been dealt with by the College. A major example occurred in 1987 when the ACR proposed developing its own relative value scale in response to the Hsiao RBRVS proposed for all of medicine. Not popular at the time, this initiative proved eventually to benefit radiologists. Self-referral, escalating in recent years, is felt to require a similar strong response, the proposal for reimbursement of imaging only to designated providers who provide high quality imaging as documented by accreditation. PMID- 17411981 TI - Quality management in radiology: historical aspects and basic definitions. AB - In today's extremely competitive economic environment, the quality management processes used by industrial companies have become commonplace at hospitals and are proving successful in improving quality and controlling costs. Continuous quality improvement (CQI) methods provide a relatively new way, compared with quality assurance (QA) and quality control (QC) methods, to improve the quality of health care. Continuous quality improvement should be considered a philosophy rather than simply a methodology; it assumes no endpoints in improvement efforts and does not attempt to replace the older concepts of QA and QC but rather to reap their benefits and take them to a higher conceptual level. Continuous quality improvement has 4 foci: (1) to determine and meet the needs of patients and customers, (2) to approach quality improvement holistically on the basis of the identification of the underlying cause of poor performance, (3) to apply fact based management and scientific methodology, and (4) to empower its practitioners to improve quality on a daily basis. Health care institutions and radiology departments use a variety of CQI systems or models, including the model of the Joint Commission on Accreditation of Healthcare Organizations, the Six Sigma model, and the Model for Business Excellence of the European Foundation for Quality Management. The International Organization for Standardization 9000, which creates a suitable organizational environment for the implementation of a CQI system, can be considered an effective QA and QC method. PMID- 17411982 TI - Radiologic quality and safety: mapping value into radiology. AB - The authors have created a radiology quality map to help understand the opportunities for improvement in the radiologic safety, reliability, quality, and appropriateness of examinations and interventions. It entails 9 steps with dozens of specific opportunities for improving care to patients. The radiology profession has an obligation to robustly document and improve quality and safety in its practice. PMID- 17411983 TI - Quality research in radiation oncology: a self-improvement initiative 30 years ahead of its time? AB - The quality of cancer care in the United States should be better than it is. Society has demanded improvement, but much work remains to be done to define and measure both the current quality of care and the steps needed to optimize such care. Various public and private organizations are directing early efforts toward attempts to determine the quality of selected oncology services as a first step in a broad-based quality improvement process. In contrast, the ACR Patterns of Care Study (PCS) for over 30 years has relied on exemplary voluntary engagement by American radiation oncologists in critical self-assessment and self improvement as a highly effective pathway to improved practice quality. This article provides an overview of the documented historical and recent impact of PCS research findings on practice and describes the deliberate adaptation of the PCS identity and methodology to the quality-sensitive national environment with the new project name Quality Research in Radiation Oncology. The article concludes with a discussion of the rationale for continuing this unique quality improvement initiative and some of the challenges to this imperative that are being faced. PMID- 17411984 TI - Health savings accounts: description, analysis, and implications. AB - Health savings accounts (HSAs) were authorized in federal legislation in 2003 and became effective January 1, 2004. The principal motive was to encourage working age persons to be more cost conscious and to make more prudent health care spending decisions, thereby restraining growth in health care spending. A secondary motive was to make it easier for individuals to accumulate funds over time to be used for paying for health care. As of March 2005, enrollment in HSAs exceeded 1 million. Contributions to HSAs can be made by individuals or employers. Neither contributions nor withdrawals for qualified health care expenses are subject to federal taxation. Unused funds can be carried over to the next year or to another employer; this feature makes HSAs more attractive than other types of health spending accounts. An HSA must be coupled with a high deductible health plan. The out-of-pocket spending gap between the annual HSA contribution and the minimum deductible is intended to make people more cost conscious in their health spending decisions. Prototypes of HSAs have been tried on a limited basis in the private sector, and these provide some lessons for the likely effects of HSAs on the socioeconomics of enrollment; the type and timing of health care spending; health outcomes; and effects on individuals, employers, and providers. PMID- 17411985 TI - Turf wars in radiology: training in diagnostic imaging: how much is enough? AB - Although many medical organizations have issued training standards for various types of diagnostic imaging, these standards tend to be arbitrary and may not adequately address the question of how much training is necessary or sufficient. In this article, the authors review the evidence pertaining to this question, which shows that acquiring expertise in imaging is a lifelong process and that there may never be such a thing as "enough" training. It is therefore likely that even though some nonradiologist physicians may meet the standards developed by some organizations, they will not have achieved the expertise that comes with long years of experience or subspecialty training in imaging. Payers and hospital credentials committees have a responsibility to patients to try to help reduce medical errors by ensuring that only those physicians who are best trained and most experienced are allowed to perform and interpret imaging studies. PMID- 17411986 TI - Selected accounts receivable performance statistics for radiology practices: an analysis of the adjusted collection percentage and days charges in accounts receivable. AB - OBJECTIVE: To provide comparative data and analysis with respect to accounts receivable management performance criteria. METHODS: Data from 3 sources were analyzed: the Radiology Business Management Association's (RBMA) 2003 Accounts Receivable Performance Survey; the RBMA's 2003 Accounts Receivable Survey; and Hogan and Sunshine's 2004 Radiology article "Financial Ratios in Diagnostic Radiology Practices: Variability and Trends," the data for which were drawn primarily from the ACR's 1999 Survey of Practices. The RBMA surveyed (via e-mail and postal mail) only its members, with response rates of 15% and 9%, respectively. The ACR's survey response rate was 66%, via postal mail, and was distributed without regard to the RBMA membership status of the practice manager or even whether the practice employed a practice manager. Comparison among the survey results provided information on trends. RESULTS: Median practice professional component adjusted collection percentage (ACP) deteriorated from 87.3% to 85.1% between the RBMA surveys. Practices limited to global fee billing faired much better when performing their billing in house, as opposed to using a billing service, with mean ACPs of 91.2% and 79.4%, respectively. Days charges in accounts receivable 2004 mean results for professional component billing and global fee billing were nearly identical at 56.11 and 55.54 days, respectively. The 2003 RBMA survey reported 63.74 days for professional component billing and 77.33 days for global fee billing. The improvement from 2003 to 2004 was highly significant for both professional component billing and global fee billing. The 2004 RBMA survey also reflected a rather dramatic improvement in days charges in accounts receivable compared with Hogan and Sunshine's results, which showed a mean of 69 days charges in accounts receivable. CONCLUSION: The conflicting trends between ACP performance and days charges in accounts receivable performance may be explained by the increasing sophistication of accounts receivable management processes (improving days charges in accounts receivable) and the deterioration in the general economy between survey periods (decreasing ACPs). Additionally, generally better accounts receivable management performance was experienced by practices employing RBMA members (RBMA survey participants) compared with those that may or may not have employed RBMA members (ACR survey participants). PMID- 17411987 TI - Research residents' choices of academic radiology careers. AB - PURPOSE: In 1993, the University of Virginia instituted a separate research residency program in the Residency Match (one resident per year). As part of the program, research residents took their second year away from clinical service for research under mentors and enrolled in a postgraduate educational program. This article evaluates what fraction of these residents eventually chose research careers. METHODS: The article reviews the career choices of 51 residents (8 research residents, 43 conventional clinical residents) who entered residency training between 1993 (the year the research residency program began) and 1999 (the last year for which postfellowship data were available) and compares the rates of research residents and conventional clinical residents choosing academic careers. RESULTS: Of the 8 research residents, 5 are currently in academic practice, but 2 of the 8 were dismissed from the program for poor clinical performance. Of the 43 clinical residents, all but 2 are in community practice; 6 of these residents either were dismissed or left the program voluntarily. Research residents had greater difficulty with board examinations than clinical residents. CONCLUSIONS: A dedicated research residency can increase the rate of trainees ultimately choosing academic careers, but there are significant risks for these trainees that they will not be able to sufficiently accommodate both clinical and research training roles. PMID- 17411988 TI - Priority setting, NIBIB style. PMID- 17411989 TI - Role of information technology in improving the quality of care in radiology: an overview. PMID- 17411990 TI - Magnetic resonance imaging guidance of ablative thermal therapies. PMID- 17411991 TI - Leo G. Rigler. PMID- 17411992 TI - On-call resident preliminary reads: should the process continue? PMID- 17411994 TI - Cliffs, marathons, and searches for the silver bullet. PMID- 17411995 TI - It takes collaboration. PMID- 17411996 TI - The statements by the American College of Cardiology on in-office cardiac imaging performed by cardiologists. PMID- 17411997 TI - Teaching the art of verbal consultation. PMID- 17411998 TI - Financing American health care: the new new idea. PMID- 17411999 TI - Open-source software opportunities and challenges. PMID- 17412000 TI - The controversy over advanced cardiovascular imaging: relative roles of radiologists, cardiologists, and other physicians in CT and MRI of the cardiovascular system. AB - PURPOSE: There is considerable conflict between radiologists and cardiologists over who has the experience and knowledge to perform computed tomography (CT) and magnetic resonance imaging (MRI) of the cardiovascular (CV) system. The purpose of this study was to determine the relative roles of the different specialties in these procedures. METHOD AND MATERIALS: The nationwide 2003 Medicare Physician Supplier Procedure Summary Master Files were used to study all 8 codes for CT angiography and all 12 codes for magnetic resonance angiography and MRI of the heart. The database provides information on the number of studies performed in each code and the specialties of the physician providers. All professional component and global claims were tabulated, but technical-component-only claims were excluded because that would have led to double counting studies. RESULTS: In 2003, in the Medicare fee-for-service population, a total of 266,142 CV CT studies were performed. Radiologists performed 257,581 (96.8%), and cardiologists performed 1,246 (0.5%). Primary care physicians (PCPs) and radiation oncologists each performed more of these studies than cardiologists: 1,879 and 1,389, respectively. Independent diagnostic testing facilities (IDTFs) and multispecialty groups together performed 2,398 (0.9%); although Medicare lists each of them as a medical "specialty," the actual specialty of a provider cannot be determined. A total of 110,743 CV MRI studies were performed. Radiologists did 100,788 (91.0%), and cardiologists did 3,257 (2.9%). In addition, IDTFs and multispecialty groups together performed 4,644, PCPs 753, and radiation oncologists 537. CONCLUSION: Cardiologists had only minimal roles in CV CT and MRI in 2003. Their procedure volume was so small that it raises a question as to whether they could be gaining meaningful experience or providing meaningful training to fellows. Radiologists strongly dominated in the use of these modalities for imaging CV structures. PMID- 17412001 TI - Medicare coverage of PET for cervical cancer. AB - Positron emission tomography (PET) with 2-[(18)F]fluoro-2-deoxyglucose (FDG) has had a major impact on the initial evaluation and follow-up of patients with cancer. Yet because Medicare coverage standards differ from the US Food and Drug Administration's approval requirements, FDG-PET has not been covered by the Centers for Medicare and Medicaid Service (CMS) for general oncologic indications. Instead, starting in 1995, CMS began to cover FDG-PET for specific cancers and indications after an individualized review of the evidence of benefit in the scientific literature. The CMS limited coverage to circumstances in which results could be expected to change patient outcomes and in which FDG-PET could either help avoid an invasive procedure or would replace other imaging studies or when the results of prior conventional imaging were inconclusive. To expand the coverage of FDG-PET for cancer imaging, the Academy of Molecular Imaging began working with various organizations to submit applications for coverage of several additional cancers. In 2003, Washington University investigators requested FDG PET coverage for the staging and restaging of cervical cancer. In January 2005, CMS issued a decision memorandum covering FDG-PET for patients with newly diagnosed cervical cancer subsequent to conventional imaging that is negative for extrapelvic metastasis. The CMS also announced a new approach for evaluating new technology and indicated its intent to cover FDG-PET for all oncologic indications as part of a national PET data registry. With this approach, FDG-PET for a presently noncovered indication will now be covered, if an attending physician completes a brief case report both before and after the PET study. The CMS is attempting to implement the registry by late 2005. PMID- 17412002 TI - Emerging technology and practice: characteristics of PET-CT technologists. AB - PURPOSE: The purpose of this study was to determine how many and which computed tomography (CT) scans are performed by technologists on positron emission tomography (PET)/CT fusion scanners. MATERIALS AND METHODS: A survey of registrants was conducted by the American Registry of Radiologic Technologists (ARRT). The sample was selected on the basis of disciplines of employment, including nuclear medicine technology (NMT), CT, and fusion. Of 676 technologists with appropriate characteristics (eg, staff technologists), a random sample of 400 was selected. A total of 173 surveys were returned (43%), and 56 respondents were identified as PET-CT technologists. RESULTS: Among PET-CT respondents (n = 56), 52% worked in community or university hospitals, 38% in free-standing centers, and 10% in mobile units. On the basis of the percentage of time (mean = 54%) and the number of patients per week (mean = 25), respondents spent most of their time in PET-CT or NMT. Of 53 CT-only procedures that form the ARRT's CT clinical experience requirements, 18 were identified as requirements respondents could fulfill (ie, they could be performed 3 to 5 times within a 24-month period). The average frequency of performance was at least 1 time in the past 6 months. Quality assurance procedures had higher endorsement rates (mean = 70%) and higher frequencies (mean = 1.7 times in 6 months). More than 35% of respondents could fulfill current CT clinical experience requirements. CONCLUSIONS: Positron emission tomography/CT technologists are mainly performing fusion scans; CT-only scans are not widely performed by PET-CT technologists on fusion scanners; PET-only scans are also rare. Most of the CT-only tasks performed are quality assurance. PMID- 17412003 TI - Health care policy consultants to the US government: who are they? and why radiologists should care. AB - Although expert staff members provide an array of assistance to our legislators and president, in setting and implementing health policy, there is always a need for further expertise. The Congressional Budget Office, the Medicare Payment Advisory Commission, the US Government Accountability Office, the Congressional Research Service, and the Institute of Medicine represent distinct areas of expertise, process, and timeliness in providing high-level research to our governmental officials. Understanding these organizations and, at times, providing them with needed external background and support can be helpful to the profession of radiology and the improvement in health services in this country. PMID- 17412004 TI - The diffusion of new imaging technologies: a molecular imaging prospective. AB - Molecular imaging is a complex of technologies that will diffuse into clinical practice over the next 10 to 20 years. Because of characteristics of both the technology and the environment, molecular imaging has the potential to be disruptive to conventional radiology practice. Environmental influences, including scientific and clinical characteristics of the technology, the interests of commercial firms, competition among radiologists and with other specialists, and regulation and reimbursement decision making will influence both the pace of diffusion and the eventual success of various molecular imaging technologies. Molecular imaging poses both threats and exciting opportunities for radiologists. Radiologists must decide how they wish to cope with the advent of molecular imaging, choosing for the present among ignoring its potential, attempting to fit molecular imaging into current practice models, or morphing how they practice to accommodate molecular imaging as a part of radiologic practice. PMID- 17412005 TI - Clinical implementation and efficiency of kilovoltage image-guided radiation therapy. AB - This paper describes measurements of clinical efficiency and time requirements associated with image-guided radiation therapy (IGRT). In June 2004, the authors' institution installed an integrated kilovoltage (kV) imaging system attached to a medical linear accelerator for radiographic target localization. Over the past year, 242 patients have been localized with the kV radiographic imaging system for a total of 2,700 fractions. Data were analyzed by reviewing the time required for each patient's IGRT session, broken into both image acquisition and image analysis time. Average IGRT procedure time was reviewed pertaining to months, treatment sessions, disease sites, and radiation therapists. Results showed that the average IGRT procedure time was reduced from 450 to 237 seconds from June 2004 to June 2005. Further analysis revealed that each therapist showed improvement in reducing the IGRT procedure time from the first month of use to the month of June 2005. The routine use of IGRT may ultimately be performed within 3 to 4 minutes, with minimal disruption to the clinical treatment process. PMID- 17412006 TI - Integrating imaging informatics into the radiology residency curriculum: rationale and example curriculum. AB - Imaging informatics, as part of the wider emerging discipline of medical informatics, remains poorly defined. However, many educators agree that formalized and flexible training in the collection, display, manipulation, storage, retrieval, and communication of imaging data, as well as the integration of these data into larger databases, should be introduced into the period of radiology residency training. The authors review the importance of such training to those individuals now preparing for clinical practice and research. They describe a sample imaging informatics curriculum that can be incorporated into a 4-year radiology residency program and the significance of such training in establishing a new subdiscipline focusing on imaging information technologies. PMID- 17412007 TI - A resident's primer of Medicare reimbursement in radiology. AB - After having completed medical school, residency, and in many cases specialized fellowships, radiologists are well equipped to practice clinical radiology. However, they receive little if any exposure to the business of radiology, such as coding, billing, and other administrative duties that maintain the financial well-being of any medical practice. Medicare insolvency, managed care, self referral, and increasing imaging by nonradiology specialists are all issues creating a competitive and ever changing medical environment, and understanding the economic and business aspects of health care is becoming increasingly important for both academic and private practice radiologists. The intent of this paper is to provide new radiologists as well as radiologists in training an introduction to the reimbursement system, as well as to provide a generalized review of the process for practicing radiologists. In particular, this article addresses the fundamentals of the Medicare fee-for-service reimbursement process as well as the factors considered in arriving at the valuation of radiologic services by Medicare. PMID- 17412008 TI - Aristotle on leadership. PMID- 17412009 TI - The new reconstruction codes: 3-D is better than no D. PMID- 17412010 TI - PACS archiving: a multivariate problem and solution. PMID- 17412011 TI - Albert Soiland. PMID- 17412012 TI - The laws of gravity. PMID- 17412013 TI - The lost keys may not lie under the lamppost. PMID- 17412014 TI - The appreciative approach to innovation and improvement. PMID- 17412015 TI - Electronic medical records: chapter one, the promise. PMID- 17412016 TI - Imaging and informatics at the National Cancer Institute, part 1. PMID- 17412017 TI - The 2005 Robert D. Moreton lecture: the inappropriate utilization of imaging through self-referral. AB - Self-referral in diagnostic imaging has adverse consequences for our health care system. It inevitably leads to overutilization. If untrained physicians try to interpret images, they will make avoidable errors. Imaging facilities operated by untrained physicians are more likely to produce poor-quality examinations. These statements are backed up by abundant evidence in the medical literature. Major payers such as Medicare and the Blue Cross Blue Shield Association are aware of the problem. There are a number of possible remedies, and it remains to be seen whether they and other payers will adopt them. PMID- 17412018 TI - 10 CFR Part 35: changes to the NRC rule governing the medical use of radioisotopes and implications for radiologic practice. AB - In the spring of 2005, the US Nuclear Regulatory Commission completed its revision of Title 10, Part 35, of the Code of Federal Regulations, its regulatory framework for the medical use of radioisotopes. Because of the complexity of the regulation and the lengthy period over which the revisions were accomplished, many radiologists remain uninformed regarding changes in the rule. This article highlights some of the more important of these changes as they relate to the practice of radiology, radiation oncology, nuclear medicine, and medical physics. PMID- 17412019 TI - An analysis of the buy-vs-lease decision. AB - This article presents a financial model to analyze the buy-vs-lease decision. The model is constructed from the perspective of a lessee with an operating lease and uses the concept of net present value, which calculates the current value of predicted cash flows in the future. Predicted cash flows of an operating lease compared with buying are presented in the model, as is the after-tax borrowing rate, the appropriate discount rate used in a model of this type. The article also discusses nonfinancial factors that may influence the buy-vs-lease decision, including the need for flexibility in working capital and the anticipated technological obsolescence of equipment. PMID- 17412020 TI - Determinants of radiologists' productivity in private group practices in California. AB - PURPOSE: The purpose of this study was to determine factors that would affect radiologists' productivity in private group practices in California. METHODS: Individual productivity data were collected for 236 private practice radiologists from 6 private radiology groups in California. The data included information on physician characteristics, facility indicators, and group practice factors that were hypothesized to affect providers' productivity. Statistical tests including chi-square testing and multivariate linear regression were used to analyze the effect of the 3 groupings of factors on the productivity of the radiologists. RESULTS: With increases in age, the number of years in practice, and the number of years in affiliation with the group practice, productivity seemed to decrease. On the other hand, productivity tended to increase if a radiologist was a shareholder, with an increase in the number of facilities served by the radiologist per day, in the proportion of imaging examinations and interventional procedures conducted, and in the proportion of those examinations stored in picture archiving and communication systems. CONCLUSIONS: To improve radiologists' productivity, group practices must invest in data-reporting infrastructure for tracking productivity, contract with outpatient imaging centers, and review group practice partnership composition and incentive models to ensure higher productivity. Future studies might consider examining the effect of other factors, such as time spent on nonclinical duties and the use of paramedical assistants in the practice. PMID- 17412021 TI - Optimizing the interpretation of CT for appendicitis: modeling health utilities for clinical practice. AB - PURPOSE: Error in radiology can be reduced by standardizing the interpretation of imaging studies to the optimum sensitivity and specificity. In this report, the authors demonstrate how the optimal interpretation of appendiceal computed tomography (CT) can be determined and how it varies in different clinical scenarios. METHODS: Utility analysis and receiver operating characteristic (ROC) curve modeling were used to determine the trade-off between false-positive and false-negative test results to determine the optimal operating point on the ROC curve for the interpretation of appendicitis CT. Modeling was based on a previous meta-analysis for the accuracy of CT and on literature estimates of the utilities of various health states. The posttest probability of appendicitis was derived using Bayes's theorem. RESULTS: At a low prevalence of disease (screening), appendicitis CT should be interpreted at high specificity (97.7%), even at the expense of lower sensitivity (75%). Conversely, at a high probability of disease, high sensitivity (97.4%) is preferred (specificity 77.8%). When the clinical diagnosis of appendicitis is equivocal, CT interpretation should emphasize both sensitivity and specificity (sensitivity 92.3%, specificity 91.5%). CONCLUSIONS: Radiologists can potentially decrease medical error and improve patient health by varying the interpretation of appendiceal CT on the basis of the clinical probability of appendicitis. This report is an example of how utility analysis can be used to guide radiologists in the interpretation of imaging studies and provide guidance on appropriate targets for the standardization of interpretation. PMID- 17412022 TI - Costs of a medical education: comparison with graduate education in law and business. AB - INTRODUCTION: The costs of graduate school education are climbing, particularly within the fields of medicine, law, and business. METHODS: Data on graduate level tuition, educational debt, and starting salaries for medical school, law school, and business school graduates were collected directly from universities and from a wide range of published reports and surveys. RESULTS: Medical school tuition and educational debt levels have risen faster than the rate of inflation over the past decade. Medical school graduates have longer training periods and lower starting salaries than law school and business school graduates, although physician salaries rise after completion of post-graduate education. CONCLUSIONS: Faced with an early debt burden and delayed entry into the work force, careful planning is required for medical school graduates to pay off their loans and save for retirement. PMID- 17412023 TI - Women as radiologists: are there barriers to entry and advancement? AB - PURPOSE: In consideration of the fact that women constitute only 25% of radiology residents, even though they constitute 45% of medical students, this study was conducted to determine if the trend of women choosing radiology as a career differs from that for other medical specialties and if there are differences on the basis of the gender of program directors or geographic location. The authors also wished to determine if constraints exist that prevent women from advancing into positions of leadership in radiology. METHOD: The percentage of women in each of the 186 radiology residency programs was compiled to determine the mean and standard deviation of women represented and from those data to examine if there were patterns of exclusion related to program size, location, or the gender of program directors. The membership and committee lists of the ACR and the Radiological Society of North America (RSNA) were examined to gauge the participation of women as leaders in these 2 organizations, as were the mastheads of Radiology and the American Journal of Roentgenology. The number of female chairs of academic departments was also examined. RESULTS: Over the past decade, the percentage of women in diagnostic radiology residencies has remained remarkably constant at or slightly above 25%. There was no discernable prejudice against women applicants by program size, location, or program director gender. In both the ACR and the RSNA, women are represented in positions of leadership approximately in proportion to their percentage in the general membership. Journal mastheads have fewer women than might be expected given the participation of women in academic radiology. There are a small but increasing number of women chairing academic radiology departments. CONCLUSION: The relatively low percentage of women in diagnostic radiology residencies is not a reflection of the gender of program directors. Women are represented in positions of influence and authority in major organizations in American radiology in proportion to the overall number of women in the organization. However, women continue to be underrepresented in radiology chair positions. Explanations must be sought for the relative unattractiveness of radiology to prospective women residents and barriers to the advancement of women in academic radiology. PMID- 17412024 TI - Medical directors of breast imaging centers: beyond films. AB - Over the past decade, breast radiologists have been increasingly asked to fulfill a new dynamic role as medical directors of breast imaging centers. To our knowledge, there are no standardized job descriptions nor defined roles and responsibilities for this position. Job descriptions are usually crafted to fit unique institutional and individual situations. To be an effective medical director of a breast imaging center, breast imagers must be more than just film readers. In this article, the authors describe the diverse roles of contemporary medical directors of breast imaging centers. PMID- 17412025 TI - Clinical decision support in radiology: what is it, why do we need it, and what key features make it effective? PMID- 17412026 TI - Gel dosimetry. PMID- 17412027 TI - Achieving satisfaction in radiology practice: advice to graduating residents. PMID- 17412028 TI - Gioacchino Failla. PMID- 17412029 TI - Chipping away at imaging. PMID- 17412030 TI - Only the paranoid survive. PMID- 17412031 TI - Stakeholder expectations for radiologists: obstacles or opportunities? PMID- 17412032 TI - An underestimated synergy: the workplace environment, staff morale, and patient satisfaction. PMID- 17412033 TI - Electronic medical records, chapter 2: the obstacles. PMID- 17412034 TI - Imaging and informatics at the National Cancer Institute, part 2. PMID- 17412035 TI - Strategic marketing: an introduction for medical specialists. AB - Marketing and branding are 2 of the most important factors for business success in the United States. They are particularly critical in service industries such as diagnostic imaging. However, in spite of their strategic importance in radiology success, a search of the peer-reviewed radiology literature reveals a paucity of published work that addresses marketing for imaging practices. In particular, there is a dearth of literature addressing the role (both direct and indirect) of radiologists in marketing efforts. In this article, the authors attempt to identify and correct some common misconceptions that physicians and other scientific and technical professionals have about marketing. Basic terms and preliminary concepts are introduced to provide a foundational understanding of the topic, allowing the interested reader to move forward and explore these critical issues in greater depth. PMID- 17412036 TI - Business plan to establish a CT colonography service. AB - The authors describe the University of Iowa Department of Radiology's business planning process to initiate a new service in computed tomographic colonography (CTC). Also known as virtual colonoscopy, CTC is a noninvasive technology that offers less risk, and potentially similar sensitivity and specificity, than conventional optical colonoscopy (OC). Although not currently covered by all insurance payers, about a year ago, the Centers for Medicare and Medicaid Services instituted temporary Current Procedural Terminology codes (Category III) for CTC. In locales where the procedure is not covered by insurers, it is likely to be sought by patients willing to pay out of pocket to undergo noninvasive cancer screening as an alternative to OC. Thus, CTC could become the preferred method of colon cancer surveillance by insurance providers in the near future. In developing the business plan, the authors reviewed pertinent scientific and clinical data to evaluate the need for and efficacy of CTC. Local market data were used to estimate patient and procedure volumes and utilization. The authors modeled financial expectations with respect to return on investment on the basis of recently reported models specific to CTC, resource requirements, and the operational impact of the new service on existing hospital and departmental clinical functions. Because there are few local providers of CTC in the authors' region, the business plan also included a publicity campaign and plan to market the new service, stimulate general public interest early, and differentiate the program as a leader in applying this unique new technology to promote cancer screening. Finally, the planning committee acknowledged and accommodated needs specific to the missions of an academic medical center with respect to research and education in designing the new service. PMID- 17412037 TI - Physician collaboration with payers: an important step on the road to optimum patient care. AB - In simpler times, patients willingly exchanged such things as food, clothing, and the fruits of their labor directly with physicians as payment for medical services rendered. Since the advent of third-party intermediaries who assumed responsibility for that payment, physicians have had a tenuous and often confrontational relationship with those intermediaries. This article discusses the rationale, principles, and methods whereby physicians can reverse the long standing trend of confrontation with payers and thus provide benefit to themselves, payers, and patients through a collaborative process of positive engagement. PMID- 17412038 TI - Radiology physician extenders and their perspective in their own words. AB - Physician assistants have been previously studied to evaluate their satisfaction with their vocation. A recent evaluation of physician assistants demonstrated common themes that indicated job satisfaction and dissatisfaction. Provider satisfaction increases the likelihood of provider retention in any practice. Physician extenders provide an enlarging portion of radiology services. Understanding the motivations, expectations, and needs of this group of midlevel care providers as perceived by these providers will allow radiology practices to better address these issues and retain these needed practitioners. Previous evaluations have been performed on the physician assistant community at large rather than on radiology extenders, a relatively new position. Therefore, the authors sought to assess the factors that influence vocational satisfaction in radiology extenders at their institution to identify issues specific to these providers. The use of qualitative data analysis allowed the physician extenders to speak for themselves. PMID- 17412039 TI - Alternative liability insurance: are you ready for a captive? AB - Liability insurance is a substantial and increasing expense to most physicians. This expense can be reduced at the cost of increased risk management and claims handling effort. However, if these efforts are not successful, there is the additional risk that actuarially determined future funding requirements will escalate. This article discusses several mechanisms that radiologists should consider when malpractice insurance expenses have increased beyond acceptable levels or when such insurance is not available at any price. The authors discuss using deductibles, self-insured retentions, and the development of one's own insurance company, that is, a captive. PMID- 17412040 TI - Conflict management and resolution. AB - When people work collaboratively, conflict will always arise. Understanding the nature and source of conflict and its progression and stages, resolution, and outcome is a vital aspect of leadership. Causes of conflict include the miscomprehension of communication, emotional issues, personal history, and values. When the difference is understood and the resultant behavior properly addressed, most conflict can be settled in a way that provides needed change in an organization and interrelationships. There are serious consequences of avoiding or mismanaging disagreements. Informed leaders can effectively prevent destructive conflicts. PMID- 17412041 TI - Financing radiology graduate medical education: today's challenges. AB - Radiology graduate medical education (GME) is exposed to huge financial challenges. First, there is a continuous increase in demand for imaging services by referring doctors and the general population, aggravating the staff shortage. Second, there has been an important decline in reimbursements. Third and probably most important is the progressive reduction of federal funds subsidizing GME. Medicare is the largest single contributor to GME. The Balanced Budget Act (BBA) of 1997 introduced reductions in Medicare payments to the major teaching hospitals calculated at $5.6 billion over the first 5 years after implementation. The BBA also brought other changes directly affecting GME. Financial changes in health care over the past decade have increased the pressure on academic institutions, which must preserve or improve the quality of training and the quality of care and manage an increased workload with fewer funds available and a narrow margin for errors. Yet the use of new technology promises to help simplify processes, decreasing workloads for residents and faculty members and increasing overall productivity, and new sources of funding have been suggested. By reviewing the financial challenges of radiologic training in today's academic centers, the authors reach the conclusion that there is still space for improving academic quality and the quality of care within current financial boundaries. But more reliable data about the specific benefits and drawbacks of having a residency program in a clinical radiology department are required. PMID- 17412042 TI - A good place to work. PMID- 17412043 TI - Deciphering the reimbursement lexicon. PMID- 17412044 TI - Cross-sectional imaging: a technology crisis upon us. PMID- 17412045 TI - Faculty development program for residents: an incentive to faculty recruitment. PMID- 17412046 TI - Juan A. del Regato. PMID- 17412048 TI - Re: "Radiology residency call: residents' perceptions of the on-call experience". PMID- 17412049 TI - An endangered species? PMID- 17412050 TI - Academic radiology and the future of the specialty. PMID- 17412051 TI - The goose and the nighthawk: a bedtime fable for young radiologists (with apologies to the Brothers Grimm). PMID- 17412052 TI - A continuum of quality in radiology. PMID- 17412053 TI - The last prescription drug benefit: the Medicare Catastrophic Coverage Act of 1988. PMID- 17412054 TI - The crisis in academic radiology: will we help ourselves? AB - The overall health of academic radiology suffers from insufficient funds and manpower. Although the largest academic programs in the country may have sufficient resources to maintain robust academic environments, one third to half of the academic radiology programs in the United States are struggling to maintain stable academic environments. The impact of an impaired academic radiology enterprise on the specialty of radiology is far reaching. As academic departments falter, the quantity and quality of research and educational programs deteriorate. In the short term, this situation makes our specialty vulnerable to predatory strikes by other specialists who covet our field; in the long term, it leads to obsolescence. Fortunately, radiology is a lucrative specialty, and we have the wherewithal to help ourselves. To ensure a vibrant future for our specialty each of us must accept an obligation to invest in our academic foundation. In particular, private practice radiologists must recognize this obligation and pledge their time and/or resources to help shore up the academic departments. PMID- 17412055 TI - Is your profession worth 1%? PMID- 17412056 TI - When does expert witness testimony constitute a violation of the ACR Code of Ethics? The role of the ACR Committee on Ethics. AB - The ACR established the Committee on Ethics in 1997. As outlined in a prior article in JACR, the committee reviews allegations that an ACR member violated the Code of Ethics. One type of complaint is that an ACR member provided biased or inaccurate expert medical testimony. This article describes how the committee reviews expert witness complaints, decides whether to investigate them, and in some cases imposes disciplinary sanctions against a member. PMID- 17412057 TI - Misrepresentation of publications among radiation oncology residency applicants. AB - INTRODUCTION: Authorship misrepresentations have been described for residency and fellowship applications for various medical specialties. This study assessed the prevalence of misrepresented publications in radiation oncology residency applications. MATERIALS AND METHODS: The authors reviewed 117 applications to their residency program for a single 2004 position offered through the National Resident Matching Program. Publications listed on the applications were verified for accuracy, with the results and applicants' demographic information recorded. RESULTS: A total of 49 applicants (42%) claimed authorship of published research citations. The number of published citations averaged 3.6 per applicant (range, 1 23). Of the applicants reporting citations, 22% (11 of 49) listed inaccurate citation information. Overall, 9% of the citations (15 of 174) were considered misrepresentations, with 9% of the total number of applicants (11 of 117) responsible for inaccurate bibliographies. There was a significant relationship of United States Medical Licensing Examination score with publication misrepresentation, in which those with scores of 235 or greater who listed publications were more than 7 times more likely to have inaccurately listed citations (odds ratio, 7.67; 95% confidence interval, 1.12-52.31; P = .04). CONCLUSION: The misrepresentation of bibliographic citations does exist among radiation oncology residency applicants. Using a comprehensive search, the authors found that 22% of those who had listed at least 1 article had misrepresented publications on their applications. PMID- 17412058 TI - Radiologist assessment of PACS user interface devices. AB - INTRODUCTION: The process of radiologic interpretation has been revolutionized with the adoption of filmless imaging. Despite the radical change in viewing techniques and a rapidly expanding array of display, navigation, and manipulation tools, the keyboard-and-mouse configuration remains the standard for user interaction during radiologic interpretation. METHOD: In this study, 6 readers evaluated 6 alternative user interface devices (UIDs), including 5-button and 8 button mice, a gyroscopic mouse, a multimedia controller, a handheld mouse-and keyboard combination device, and a gaming joystick. Each participant assessed each device during the real-time daily imaging interpretation of magnetic resonance, computed tomographic, and general x-ray studies over a 2-week period and completed a detailed questionnaire on the ease of use, comparative utility as an alternative device to mouse and QWERTY keyboard, efficiency, workflow, and the ease of customized programming. RESULTS: No single device was completely able to replace the mouse and keyboard in the estimation of participants, and the 5 button mouse was preferred over the 8-button mouse, although several participants noted that this might be a function of learning curves that exceeded the 2-week study period for each device. Several specific functionalities of the alternative UIDs were praised by users, who also suggested the possibility of combining these alternative devices for 2-handed use. CONCLUSION: Consideration should be given to the potential for the increased acceptance of and efficiency with alternative UIDs that can be integrated into picture archiving and communications system (PACS) workstations. Additional studies are planned to use software applications to evaluate different UIDs at PACS workstations. PMID- 17412059 TI - Strategic marketing, part 2: the 4 P's of marketing. AB - Marketing and branding are critical business functions that are often ignored or misapplied in the health care sector. Radiology professionals are facing unprecedented competition, turf battles, and other pressures. One tool that can help in meeting this onslaught is to improve your marketing efforts. Some of the most expensive mistakes in marketing (medical and otherwise) are caused by not paying attention to the basics. These "rookie" errors can be avoided by a careful review of the 4 key principles of introductory marketing: product, price, placement, and promotion. This article reviews these concepts as they relate to medical marketing. PMID- 17412060 TI - Considerations of cost-effectiveness for new radiation oncology technologies. AB - PURPOSE: The additional equipment and personnel costs of supplying image-guided radiation therapy (IGRT) technology have caused many to question if the marginal gains in patients' health-related quality of life are worth the additional cost. Novel IGRT technologies, including cone-beam computed tomography and helical tomotherapy, provide the opportunity to study cost and effectiveness for patients. MATERIALS AND METHODS: This methodologic study proposes to evaluate the cost and effectiveness of treating conventional radiotherapy versus IGRT patients prospectively among several institutions. The cost of treating patients varies among institutions depending on personnel, equipment, and overhead costs, but the nature and quality of services provided are expected to be consistent. RESULTS: The study will track cost information at a single institution and simultaneously as the median from multiple institutions. Effectiveness measures will include both standard quality-adjusted life-year instruments completed by patients and performance status measures completed by institutional personnel. In addition, disease-specific effectiveness measures will be accommodated in the study. Each participating institution will use the same effectiveness measures to track patients with similar diseases. CONCLUSION: The resulting cost and effectiveness data will be available to investigators at any point during the study, immediately on the completion of a trial, or when statistical acceptability is achieved. These considerations are being incorporated into a high-level information model under development. PMID- 17412061 TI - Multiple PIs: a win for imagers and team science. PMID- 17412062 TI - Establishment of an interventional radiology clinic to follow untreated abdominal aortic aneurysms. PMID- 17412063 TI - Advanced image processing in the clinical arena: issues to consider. PMID- 17412064 TI - Maintenance of certification in radiologic physics. PMID- 17412065 TI - Earl R. Miller. PMID- 17412066 TI - Re: "the nighthawk: bird of paradise or albatross?". PMID- 17412068 TI - Strategies for the future. PMID- 17412069 TI - Doing well by doing good: the ACRIN Fund for Imaging Innovation. PMID- 17412070 TI - Radiologists and cardiologists should work together on advanced cardiac imaging. PMID- 17412072 TI - Health care reform: the conservative agenda. PMID- 17412073 TI - Imaging biomarkers and the future of radiology. PMID- 17412074 TI - Training for the future of radiology: a report of the 2005 Intersociety Conference. AB - The field of radiology has expanded dramatically and now encompasses a broad range of imaging examinations and image guided procedures. These imaging technologies are powerful tools which provide valuable information, and combining modalities further enhances their value. The changes our imaging technologies have brought bring into question our training methods, especially the value of the clinical year prior to entering radiology. As the quality of our health care systems is being challenged, demonstration of both quality processes and outcomes are needed. Maintenance of Certification, which demonstrates continuing learning and practice improvement, has become an important part of a radiologist's quality credentials. PMID- 17412075 TI - Professionalism and physician interactions with industry. AB - Many gifts from industry to physicians and medical organizations serve important and beneficial functions. However, the spiraling cost of health care is resulting in increasing governmental and medical scrutiny of legal and ethical issues concerning relationships between commercial companies and physicians. Professional oversight bodies are updating standards concerning the ethical responsibilities of physicians. This article presents a broad framework for understanding the professional and legal responsibilities of physicians when interacting with industry. Physicians have unique responsibilities based on the "fiduciary" nature of the patient-physician relationship and specified laws regarding health care. Physicians must protect the best interests of patients, with clinical decisions free of undue influence. Physicians have special obligations related to receiving gifts from industry or receiving payments that may be construed as "kickbacks" or fraud. To ensure that gifts do not compromise (or seem to compromise) professional judgment, physicians should generally not accept personal gifts from industry and consider accepting only those that primarily entail a benefit to patients, are not of substantial value, and have no "strings" attached. After reading this article, the reader should be able to describe the impact gifts have on physicians' behavior, the privileges and obligations of physicians, and conflicts between professional obligations and personal gifts. PMID- 17412076 TI - Radiology and industry: can this relationship be saved? PMID- 17412077 TI - Comparison of productivity and cost of full-time and part-time faculty members in an academic department of radiology. AB - PURPOSE: To compare financial productivity and employment costs of full-time and part-time faculty members and to assess qualitative factors related to part-time employment. MATERIALS AND METHODS: Financial productivity and employment costs were compared for full-time and part-time staff members on the basis of a review of financial records from fiscal year 2005. Part-time and full-time faculty members were interviewed to determine reasons for working part-time and to assess the attitudes of both full-time and part-time faculty members toward part-time status. RESULTS: Part-time radiologists working an average 65% schedule constituted 13% of the total full-time equivalent faculty complement and performed 14% of the clinical work at an average of 0.88 physicians' component total relative value units while more than covering their costs. Part-time radiologists were more productive while interpreting examinations of lesser relative value unit value than full-time radiologists. CONCLUSION: Part-time radiologists are an important resource for the specialty and should be nurtured to stay in practice. PMID- 17412078 TI - To err is human, to correct divine: the emergence of technology-based communication systems. AB - An increasing emphasis has been placed on the importance of timely communication of imaging results, especially to the extent that clinical management decisions are modified by the information. Various methods of transmitting results have been proposed and developing technology can now be applied to helping to ensure the timely receipt of such results in a busy clinical environment. Stratifying levels of urgency, ensuring redundancy of potential recipients of such information, and the ability to assess desired benchmarks are objectives that involve many stakeholders, including radiologists, treating physicians, and institutions. An enterprise approach to this challenge, including commercially available systems, offers a potentially cost-effective solution that addresses both risk management and quality improvement goals. PMID- 17412079 TI - 300,000,000 customers: patient perspectives on service and quality. AB - Radiologists have always been dedicated to service and quality in imaging. In the past few years, many entities, mostly external to the profession itself, have entered the business of defining, measuring, and reporting both service and quality. There is a great deal at stake in how this unfolds, and there are many potential stakeholders in both the process and the outcome. This article addresses the perspectives of patients on service and quality issues. In particular, it is important for those in the profession to understand that their customers often view their services very differently than those within the business. The author addresses several of the most common and important perceptual issues that practitioners need to address in understanding these disparities. Success or failure will determine how satisfied patients are; how they fill out scorecards and report cards; and what they say about practitioners to their family, their neighbors, and perhaps 2 billion of their closest friends on the Web. Ultimately, how well radiologists address these issues will determine a substantial component of pay for performance, their ability to contract, and the individual decisions of patients and their families to come to them at all. PMID- 17412080 TI - Utilization management in radiology: basic concepts and applications. AB - With the current constraints on health care resources and emphasis on value for money, resource utilization must be constantly revised and its value demonstrated. For this purpose, utilization management (UM) provides a series of methods to evaluate the appropriateness of health care resources. The ultimate goal of UM is to achieve the best patient outcomes using the most appropriate resources. The implementation of these procedures also targets variation in practice and promotes cost-effective clinical decision making. Inappropriate utilization is a major problem in health care today, particularly in capital intensive fields such as imaging. High imaging utilization supposes losses to insurers and care-provider organizations and an overload for radiologists, while underutilization carries significant risks for patients, with the potential for a negative impact on outcomes. Imaging use must be assessed as frequently as new technologies arise. Given the fast pace at which this is happening, UM provides a crucial solution for continuous revision and assessment as an integrative process. The authors offer a practical guide for radiologists in UM, describing its role in today's health care finances. The key issue is to present UM as an approach for improvement that could bring benefits to all parties by adding efficiency to the health care delivery process. Utilization management represents an important opportunity for radiologists to be involved in the decision-making process, regulating the use of their services and providing fast feedback to other specialties that rely on imaging as a complement to their clinical practice. PMID- 17412081 TI - Electronic evaluation systems for radiology residency and fellowship. AB - Electronic evaluation systems are becoming more commonplace in radiology residencies. This article describes the authors' experience in the development of such a system and a method that a residency program or graduate medical education office could use to evaluate commercially available programs. PMID- 17412082 TI - A new kind of intelligence? PMID- 17412083 TI - The ordering physician rules! PMID- 17412084 TI - Respiratory gating for radiotherapy. PMID- 17412085 TI - Are we ready to be cardiac imagers? PMID- 17412086 TI - Clinical teleradiology in Australia: practice and standards. PMID- 17412087 TI - George Winslow Holmes. PMID- 17412088 TI - Looking to the future. PMID- 17412090 TI - Training a better radiologist. PMID- 17412091 TI - Wither primary care? PMID- 17412092 TI - Quantum grants: request for applications from the NIBIB. PMID- 17412094 TI - History of research in medical image perception. AB - Human observers engage in 2 interrelated processes when interpreting medical images: perception and analysis. Perception is the unified awareness of the content of a displayed image that is present while the stimulus is on. Analysis is determining the meaning of the perception in the context of the medical problem that initiated the acquisition of the image. Radiologists have, correctly, regarded image analysis as their primary field of research. They have naively assumed that what they perceive in images is a faithful representation of the images' information content and have not been concerned with perception unless it fails. Failures have stimulated research on quantifying observer performance, defining image quality, and understanding perceptual error. This article traces the historical development of the use of receiver operating characteristic analysis for describing performance, the development of signal-to noise ratio psychophysical models for defining task-dependent image quality, studies of error in small lesion detection, and the beginnings of studies of the nature of expertise in image interpretation. The history is traced through published articles. PMID- 17412095 TI - The place of medical image perception in 21st-century health care. AB - An image that is not perceived and interpreted can have no positive impact on health care. In this article, the authors review publicly available data and the published literature concerning the unitary event of the perception and interpretation of medical images. Their review shows that this event occurs as frequently as do major medical, public health, and public safety events in the United States; constitutes a significant economic activity; and makes up a significant portion of hospital-based health care in the United States. Yet despite its central importance to the economy and to health care, the authors' analysis found that research in the perception and interpretation of medical images has been awarded minimal support by National Institutes of Health extramural funding: fewer than 5% of all National Institutes of Health-funded grants related to radiology during the 10-year period from 1994 to 2003 focused on human perception and interpretation. The increased funding of medical image perception and interpretation research could lead to important improvements in overall health care thanks to the pervasive and vital role imaging plays in modern medicine. PMID- 17412096 TI - Receiver operating characteristic analysis: a tool for the quantitative evaluation of observer performance and imaging systems. AB - Receiver operating characteristic (ROC) analysis provides the most comprehensive description of diagnostic accuracy available to date, because it estimates and reports all of the combinations of sensitivity and specificity that a diagnostic test is able to provide. After sketching the 6 levels at which diagnostic efficacy can be assessed, this paper explains the conceptual foundations of conventional ROC analysis, describes a variety of indices that can be used to summarize ROC curves, and describes several forms of generalized ROC analysis that address situations in which more than 2 decision alternatives are available. Key issues that arise in ROC curve fitting and statistical testing are addressed in an intuitive way to provide a basis for judging the validity of ROC studies reported in the literature. A list of sources for free ROC software is provided. Receiver operating characteristic methodology has reached a level of maturity at which it can be recommended broadly as the approach of choice for radiologic imaging system comparisons. PMID- 17412097 TI - Measuring observer performance in chest radiology: some experiences. AB - All decisions made under conditions of uncertainty have error rates. All meaningful decisions are made under conditions of uncertainty. Can this uncertainty be measured? Can variations in how different observers deal with this uncertainty be ascertained? The ability to measure observer performance in diagnostic imaging was one of the issues that initiated the field of medical decision analysis. This article exemplifies an approach and is worth discussing as a preamble to presenting our long-term project of measuring variations in observer performance. The paper focuses on the interpretation of chest x-ray images, although the principles and findings described can be applied to nearly every radiologic modality and interpretation task. PMID- 17412098 TI - Technology and perception in the 21st-century reading room. AB - Radiology reading rooms have changed dramatically over the past 15 years, moving from analog-light-box-based environments to digital-display-based environments. Most of the focus in the early stages of this transition was on the technology, but it soon became obvious that it was not possible or even prudent to consider the technology without considering radiologists. The information being presented to radiologists in digital reading rooms is in many ways very different from that presented on traditional film. On one hand, the digital workstation display medium itself is very different from traditional film images hung on light boxes. On the other hand, without large-area light boxes, images such as those from computed tomography (CT) can no longer be displayed all at once in a series of film sheets. The digital world also introduces the possibility of manipulating image data in ways that were never possible with analog film. Not only can radiologists manipulate image data with various image-processing tools, but also, computers can analyze images and provide even more information to incorporate into the interpretation process. As a consequence of these differences, it has been necessary to focus attention on radiologists to discover ways to optimize the digital reading environment with respect to the human visual system and the way the eye-brain system processes information. This article reviews some of the important perceptual issues that have arisen in the digital reading rooms of the 21st century. PMID- 17412099 TI - Paradigms of perception in clinical practice. AB - Display strategies for medical images in radiology have evolved in tandem with the technology by which images are made. The close of the 20th century, nearly coincident with the 100th anniversary of the discovery of x-rays, brought radiologists to a new crossroad in the evolution of image display. The increasing availability, speed, and flexibility of computer technology can now revolutionize how images are viewed and interpreted. Radiologists are not yet in agreement regarding the next paradigm for image display. The possibilities are being explored systematically through the Society for Computer Applications in Radiology's Transforming the Radiological Interpretation Process initiative. The varied input of radiologists who work in a large variety of settings will enable new display strategies to best serve radiologists in the detection and quantification of disease. Considerations and possibilities for the future are presented in this paper. PMID- 17412100 TI - Reader variability: what we can learn from computer-aided detection experiments. AB - Radiologists interpreting images vary in their identification of disease. When 1 radiologist reinterprets the same case differently, it is called intraobserver variability; when 2 radiologists differ with each other on a case, it is called interobserver variability. Computer-aided detection (CAD) systems can increase the detection of disease. When radiologists use these systems, they are aided in the detection of lesions that they might have detected had they reinterpreted the images or that other radiologists might have identified had they interpreted the images without CAD. This article demonstrates how most cases "newly" identified by radiologists working with CAD are actually cases that the radiologists or other radiologists would have identified had they interpreted the images without CAD. Computer-aided detection, therefore, decreases intraobserver variability and interobserver variability. PMID- 17412101 TI - Design and ergonomic considerations for the filmless environment. AB - Planning and executing the redesign of a traditional institutional radiology reading room to conform to the radically different requirements of digital imaging are reviewed, with examples drawn from the authors' experience and from the growing body of literature on this subject. Included are best-practice recommendations and real-life examples on initial design and planning, stakeholder involvement, identifying and hiring consultants, architectural planning, the designation of a radiology point person, rethinking room and workstation design, the selection of ergonomic furniture and fittings, identifying optimal environmental elements, fine tuning and lessons learned, and going digital. PMID- 17412102 TI - New opportunities in computer-aided diagnosis: change detection and characterization. PMID- 17412103 TI - Pregnant patient. PMID- 17412104 TI - Communication between radiologists and patients: an unsolved issue. PMID- 17412105 TI - Francis H. Williams and William H. Rollins. PMID- 17412106 TI - Developing future leaders and leadership training in radiology. PMID- 17412108 TI - Identifying and helping struggling residents. PMID- 17412109 TI - Physician performance and congressional hardball. PMID- 17412110 TI - Translational research, the NIH, and radiologists. PMID- 17412111 TI - Information overload. PMID- 17412112 TI - ACR Appropriateness Criteria on incidentally discovered adrenal mass. AB - For patients without a history of malignancy, most small (less than 3 cm) incidentally discovered adrenal masses are benign, and extensive and costly workup is usually not justified. For incidentalomas from 3 to 5 cm in size, computed tomography, magnetic resonance imaging, 2-[(18)F]fluoro-2-deoxyglucose positron emission tomography, adrenal biopsy, or surgery can be considered. Lesions larger than 5 cm should be removed because of the higher risk for malignancy. For patients with histories of malignancy, incidentally discovered adrenal masses are more often malignant, and thus even smaller adrenal lesions are suspect. Adrenal biopsy should be reserved for cases in which the results of noninvasive techniques are equivocal. PMID- 17412113 TI - Evidence-based radiology and the ACR Appropriateness Criteria. AB - The ACR Appropriateness Criteria are an important effort aimed at establishing when imaging should be used in medicine. However, substantial improvements in the Appropriateness Criteria are necessary for them to meet current standards for evidence-based guidelines. In this article, we provide concrete suggestions for improving the evidence basis of the Appropriateness Criteria. PMID- 17412114 TI - The ACR Appropriateness Criteria: view from the committee chair. PMID- 17412115 TI - Evidence-based radiology. AB - The role of evidence-based medicine, defined as the integration of best research evidence with clinical expertise and patient values, is becoming increasingly important in health care. Evidence-based practice has 5 basic steps that can be applied to any clinical discipline: (1) formulate an answerable question, (2) find the best current evidence, (3) appraise critically, (4) apply findings to practice, and (5) evaluate performance. Evidence-based radiology (EBR) is a relatively new approach to the practice of radiology based on the principles of evidence-based medicine. Through EBR, radiologists can regularly update their knowledge, deepen their understanding of research methodologies, and use research data in clinical settings more effectively; for patients, EBR ensures that they receive the best current care. Evidence-based radiology can be used to decrease the variability in radiology practice and thus avoid the unnecessary waste of resources. PMID- 17412116 TI - Wireless remote control of clinical image workflow: using a PDA for off-site distribution and disaster recovery. AB - This paper describes a picture archiving and communication system (PACS) tool based on Web technology that remotely manages medical images between a PACS archive and remote destinations. Successfully implemented in a clinical environment and also demonstrated for the past 3 years at the conferences of various organizations, including the Radiological Society of North America, this tool provides a very practical and simple way to manage a PACS, including off site image distribution and disaster recovery. The application is robust and flexible and can be used on a standard PC workstation or a Tablet PC, but more important, it can be used with a personal digital assistant (PDA). With a PDA, the Web application becomes a powerful wireless and mobile image management tool. The application's quick and easy-to-use features allow users to perform Digital Imaging and Communications in Medicine (DICOM) queries and retrievals with a single interface, without having to worry about the underlying configuration of DICOM nodes. In addition, this frees up dedicated PACS workstations to perform their specialized roles within the PACS workflow. This tool has been used at Saint John's Health Center in Santa Monica, California, for 2 years. The average number of queries per month is 2,021, with 816 C-MOVE retrieve requests. Clinical staff members can use PDAs to manage image workflow and PACS examination distribution conveniently for off-site consultations by referring physicians and radiologists and for disaster recovery. This solution also improves radiologists' effectiveness and efficiency in health care delivery both within radiology departments and for off-site clinical coverage. PMID- 17412117 TI - Challenges to radiology resident education in the new era. AB - Over the past decade, there have been many developments that have changed the practice of radiology and the education of radiology residents. These include workforce issues, the institution of the Accreditation Council for Graduate Medical Education duty-hours restrictions, the increased use of night float systems, and the implementation of picture archiving and communication systems as well as voice recognition. This article reviews the impact on resident education, summarizes potential problems introduced by these changes, and examines proposed solutions. PMID- 17412118 TI - Turf wars in radiology: privileging and site accreditation programs--what they have accomplished for commercial health plans. AB - In recent years, some commercial health plans have instituted privileging and site accreditation programs for providers of diagnostic imaging. This article describes how these programs work and points out that they can help restrict self referral and the overutilization that accompanies it. Another advantage is that they can help eliminate low-quality providers. Radiologists should encourage health plans in their area to develop and implement these measures. PMID- 17412119 TI - The impact of professional turf battles on radiology resident education: perspectives from the radiology class of 2005. AB - PURPOSES: To assess residents' exposure to professional turf battles, via a survey of the radiology residency class of 2005, and to assess perceptions about the causes, solutions, and impact of turf battles on resident education. METHODS: An electronic survey composed by the ACR Resident and Fellow Section was completed by 168 senior residents (a 18.9% response rate) enrolled in allopathic radiology residency programs from across the United States. RESULTS: The majority of respondents (153 of 168, 91.1%) perceive a loss of professional turf from nonradiologists. Areas most often perceived as being affected by lost professional turf were interventional radiology (136/168, 81.0%), cardiothoracic imaging (106 of 168, 63.1%), and ultrasound (88 of 168, 52.4%). The sense of lost professional turf was most often attributed to inadequate residency training in the affected subspecialities (87 of 168, 51.8%). The most commonly cited motivation for ongoing turf battles was reimbursement for diagnostic imaging and image-guided procedures as a motivator for nonradiologists. The majority of residents feel that the most effective ways to counter professional turf battles include the promotion of national physician-training standards for diagnostic imaging and image-guided therapies, the mandatory accreditation of all imaging facilities, and the development of performance standards that providers of imaging services must meet to qualify for reimbursement. CONCLUSIONS: The majority of resident respondents feel that professional turf battles have had a significant impact on their education. Methods recommended to counter professional turf battles mirror recent proposals by the ACR to support mandatory national quality, safety, and training standards for providers of diagnostic imaging. PMID- 17412120 TI - Quality improvement in radiology: white paper report of the Sun Valley Group meeting. AB - The Sun Valley Group is an informal assembly of individuals interested in improving quality in radiology. Its first meeting was held in September 2005. The purposes of the meeting was to share quality improvement experiences, consider a strategy for promoting quality improvement initiatives across the radiology profession, and initiate quality benchmarking efforts. Representatives from private practice, academia, national quality programs, and international societies were in attendance. Four main themes were presented: the sharing of leading quality activities in radiology, the future of pay-for-performance systems, programs and future initiatives of professional radiology societies, and health services research guidelines for developing outcome metrics. This white paper summarizes information presented in each of these thematic areas and concludes with the group's plans for future activities. Among these is a formal educational program for all radiologists interested in implementing a quality improvement program within their practice, to be hosted by the ACR. PMID- 17412121 TI - Completely limited coding. PMID- 17412122 TI - Radiology order entry: features and performance requirements. PMID- 17412123 TI - Radiology utilization rounds of the Massachusetts General Hospital: a win-win situation. PMID- 17412124 TI - Coronary CT angiography. PMID- 17412125 TI - Vincent W. Archer. PMID- 17412126 TI - Radiology and the health care reform debate. PMID- 17412128 TI - Radiology: going, going, gone. PMID- 17412129 TI - The Massachusetts Health Care Reform Plan. PMID- 17412130 TI - Nanotechnology molecular medicine and radiology. PMID- 17412131 TI - Performing a test in an office setting without an order. PMID- 17412132 TI - Digital mammography: what next? PMID- 17412133 TI - Technology evaluation center assessment synopsis: full-field digital mammography. AB - Full-field digital mammography (FFDM) is proposed as an alternative to screen film mammography (SFM). The ability to separate and optimize the acquisition, storage, and display of images may allow greater visualization of breast cancers at equal or lower radiation doses, especially in younger women and those with denser breasts. This is a synopsis of a systematic review by the Blue Cross Blue Shield Association Technology Evaluation Center. This updated systematic review primarily incorporated the results of the ACR Imaging Network(R) Digital Mammographic Imaging Screening Trial (DMIST), which provided results on 42,760 asymptomatic women who underwent both FFDM and SFM and showed with reasonable certainty that there was no difference in the accuracy of the 2 modalities for asymptomatic women in general, with some advantages of FFDM in certain subgroups. There were no strong, new studies on the use of digital mammography compared with film mammography in a diagnostic population. However, the DMIST results indicated that tumors detected by FFDM, but not by SFM, were likely to be invasive carcinomas or medium-grade to high-grade ductal carcinoma in situ. On the basis of the suppositions that these are the cancers of greatest interest and the ones more likely to be found in a diagnostic population and that the diagnostic population may be younger on average than the screening population, it was concluded that there is sufficient evidence to support the use of FFDM for diagnostic purposes. PMID- 17412134 TI - Image quality in digital mammography: image acquisition. AB - This paper on digital mammography image acquisition is 1 of 3 papers written as part of an intersociety effort to establish image quality standards for digital mammography. The information included in this paper is intended to support the development of an ACR guideline on image quality for digital mammography. The topics of the other 2 papers are digital mammography image display and digital mammography image storage, transmission, and retrieval. The societies represented in compiling this document were the Radiological Society of North America, the ACR, the American Association of Physicists in Medicine, and the Society for Computer Applications in Radiology. These papers describe in detail what is known to improve image quality for digital mammography and make recommendations about how digital mammography should be performed to optimize the visualization of breast cancers. Through the publication of these papers, the ACR is seeking input from industry, radiologists, and other interested parties on their contents so that the final ACR guideline for digital mammography will represent the consensus of the broader community interested in these topics. PMID- 17412135 TI - Storage, transmission, and retrieval of digital mammography, including recommendations on image compression. AB - This paper on digital mammography image storage, retrieval, and transmission is 1 of 3 papers written as part of an intersociety effort to establish image quality standards for digital mammography. The information included in this paper is intended to support the development of an American College of Radiology (ACR) guideline on image quality for digital mammography. The topics of the other 2 papers are digital mammography image acquisition and digital mammography image display. The societies that were represented in compiling this document were the Radiological Society of North America, the ACR, the American Association of Physicists in Medicine, and the Society for Computer Applications in Radiology. These papers describe in detail what is known to improve image quality for digital mammography and make recommendations about how digital mammography should be performed to optimize the visualization of breast cancers using this imaging tool. Through the publication of these papers, the ACR is seeking input from industry, radiologists, and other interested parties on their contents so that the final ACR guideline for digital mammography will represent the consensus of the broader community interested in these topics. PMID- 17412136 TI - Digital mammography image quality: image display. AB - This paper on digital mammography image display is 1 of 3 papers written as part of an intersociety effort to establish image quality standards for digital mammography. The information included in this paper is intended to support the development of an American College of Radiology (ACR) guideline on image quality for digital mammography. The topics of the other 2 papers are digital mammography image acquisition and digital mammography image storage, transmission, and retrieval. The societies represented in compiling this document were the Radiological Society of North America, the ACR, the American Association of Physicists in Medicine, and the Society for Computer Applications in Radiology. These papers describe in detail what is known to improve image quality for digital mammography and make recommendations about how digital mammography should be performed to optimize the visualization of breast cancers using this imaging tool. Through the publication of these papers, the ACR is seeking input from industry, radiologists, and other interested parties on their contents so that the final ACR guideline for digital mammography will represent the consensus of the broader community interested in these topics. PMID- 17412137 TI - You should eliminate paper from your PACS workflow: why and how? PMID- 17412138 TI - CT dosimetry--an enigma surrounded by a conundrum. PMID- 17412139 TI - Diagnosis of pulmonary embolism. PMID- 17412140 TI - New developments in the EU with regard to digital image management. PMID- 17412141 TI - Ulrich Henschke. PMID- 17412143 TI - Invention to practice. PMID- 17412144 TI - Ultrasound: is there a future in diagnostic imaging? PMID- 17412145 TI - Quality improvement organizations and specialty hospitals: a progress report. PMID- 17412146 TI - Pay for performance in radiology: ACR white paper. AB - During the next few years, some portion of physician reimbursement will be increasingly based on the quality and efficiency of service, a practice commonly referred to as pay for performance (P4P). Performance benchmarks are the discrete parameters of structure, process, or outcome metrics whose attainment defines good quality care. Private payers are already rewarding primary care physicians for practices that adhere to quality standards, are efficient, involve information technology, and result in high patient satisfaction. The Centers for Medicare and Medicaid Services will have completed the development of performance measures to be used in Medicare payment strategies for all specialties by the end of 2006 and anticipates phasing in the program fully by 2008. This article describes P4P, its importance to the ACR, the organizations involved in developing it, the ACR's activities to date, and the steps the ACR must take to ensure that radiologists are remunerated fairly as physician payment becomes based, in part, on performance. PMID- 17412147 TI - ACR practice guideline for the performance and interpretation of cardiac magnetic resonance imaging (MRI). AB - Magnetic resonance imaging (MRI) is an established imaging modality, recognized for its value in the assessment and monitoring of a wide range of cardiac pathology. It can provide physiologic as well as anatomic information. Image interpretation requires both well-developed MRI skills and knowledge of cardiac pathology. Radiologists, because of their extensive experience in MRI, have an important role in its application in the heart. The guidelines presented here are an educational tool designed to assist practitioners in providing the best possible patient care via the diagnostic methods of cardiac MRI. American College of Radiology requirements for physicians and personnel performing and interpreting cardiac MRI, which will become applicable by July 1, 2008, are also presented. PMID- 17412148 TI - ACR practice guideline for the performance and interpretation of cardiac computed tomography (CT). AB - Cardiac computed tomography (CT) is an evolving modality that includes a variety of examinations to assess the anatomy and pathology of the cardiac chambers, valves, myocardium, coronary arteries and veins, pericardium, aortic root, and central great vessels. The development of multidetector CT scanners with increasing numbers of detector rows, narrow section thicknesses, increasing scanner speeds, the ability for electrocardiographic gating, and radiation dose modulation allows the performance of CT coronary arteriography. Computed tomography coronary arteriography enables the assessment of multiple types of cardiac pathology, including intraluminal coronary arterial plaque formation, coronary artery stenosis, congenital anomalies, coronary artery aneurysms, sequelae of cardiac ischemia, and the assessment of prior vascular interventions, while providing information about cardiac and valvular function. Noncardiac structures included in cardiac CT examinations must also be evaluated. This guideline attempts to maximize the probability of detecting cardiac abnormalities with cardiac CT. American College of Radiology requirements for physicians and personnel performing examinations are also addressed and will become applicable by July 1, 2008. PMID- 17412149 TI - ALARA in pediatric interventional and fluoroscopic imaging: striving to keep radiation doses as low as possible during fluoroscopy of pediatric patients--a white paper executive summary. AB - Pediatric patients might be as much as 10 times more radiosensitive than adults. Thus, adherence to the principle of "As low as reasonably achievable" (ALARA) represents a practice mandate that minimizes ionizing radiation exposure while optimizing imaging results. This symposium is the third multidisciplinary program that focused on the ALARA principle in pediatric imaging and addressed issues associated with pediatric fluoroscopy and interventional imaging techniques. PMID- 17412150 TI - Survival strategies for radiology: some practical tips on how to reduce the risk of being sued and losing. AB - The average indemnification for radiologists has tripled in the past 15 years, making risk management in radiology necessary to reduce the threat of being sued and losing. More than 75% of radiology lawsuits allege failure to diagnose or failure to communicate in a timely manner: ultimately, one third of these lawsuits are lost by the radiologist. A failure to communicate results in a timely manner is one of the greatest problems facing radiologists today, and it is a causal factor in 80% of lawsuits. Although rarely the primary factor for these suits, this is the one area in medical malpractice litigation in which radiologists can play a crucial role in reducing risk and liability. This article provides some practical tips on how to reduce the risk for being sued and losing. PMID- 17412151 TI - From medical invention to clinical practice: the reimbursement challenge facing new device procedures and technology--part 1: issues in medical device assessment. AB - Although reimbursement shapes medical practice, it is hard to get a handle on this issue because reimbursement requirements seem to change and evolve over time, and different insurers approach it in different ways for different technologies. This paper and the 2 that follow attempt to explain the reimbursement challenges facing new medical device technology. This first paper addresses the particular characteristics of medical device innovation and explains how they complicate the technology assessment process. It also notes the importance of codes as a means to identify new technologies and procedures, citing some issues that affect technology adoption. The subsequent papers delve into specific coverage and payment matters affecting medical devices. The perspective presented in these papers is that of a medical device manufacturer, and the focus tends to be on Medicare, given the size of this insurance program and its impact on the US health care system, but the issues that are raised affecting medical innovation can be extended to all payers. PMID- 17412152 TI - Clinical indicators of radiographic findings in patients with suspected community acquired pneumonia: who needs a chest x-ray? AB - PURPOSE: To develop a prediction rule for the use of chest radiographs in evaluating for community-acquired pneumonia (CAP) based on presenting signs and symptoms. PATIENTS AND METHODS: Adult patients with acute respiratory symptoms and positive chest radiographic results from October 2004 through April 2005 were enrolled as positive cases (n = 350). An equal number of age-matched controls with acute respiratory symptoms but negative radiographic results were included. Data analyses were performed on the 6 most common individual clinical indicators (cough, sputum production, fever, tachycardia, tachypnea, and abnormal physical examination results). Additional analyses were performed for any vital sign abnormality and for the presence of vital sign or physical examination abnormalities. RESULTS: The data show that vital sign and physical examination findings are useful screening parameters for CAP, demonstrating a sensitivity of 95%, a specificity of 56%, and an odds ratio of 24.9 [corrected] in the presence of vital sign or physical examination abnormalities. In light of these results, the authors developed a prediction rule for low-risk patients with reliable follow-up, which states that chest radiographs are unnecessary in the presence of normal vital signs and physical examination findings. CONCLUSION: The data suggest that chest radiographs are unnecessary in patients with acute respiratory symptoms who present with normal vital signs and physical examination findings. Because approximately 5% of cases would be missed, however, these criteria are useful only for patients with reliable follow-up and a low likelihood of morbidity if CAP is not detected initially. PMID- 17412153 TI - Current practice patterns and recent trends in breast biopsy among radiologists and surgeons. AB - PURPOSE: To ascertain the relative roles of radiologists and surgeons in breast biopsy. METHODS AND MATERIALS: The Nationwide Medicare Physician Supplier Procedure Summary Master Files for 1999 to 2004 were used to study the 4 Current Procedural Terminology, Fourth Edition, codes for breast biopsy. Using the Medicare specialty codes, physician providers were categorized as radiologists, surgeons, or others. Procedure volumes for each of the 3 groups were tabulated for each year of the study, and trends were assessed. RESULTS: Between 1999 and 2004, the breast biopsy utilization rate per 100,000 Medicare beneficiaries increased by 43%. In 2004, a total of 144,697 breast biopsies were performed, of which 124,423 (86%) were done with imaging guidance. From 1999 to 2004, procedure volume increased by 101% among radiologists, compared with 32% among surgeons. By 2004, radiologists performed 63% of all breast biopsies, compared with 32% by surgeons and 5% by others. Within the 2 codes that are specific for the use of imaging guidance, radiologists performed 72%. CONCLUSIONS: Radiologists predominate in breast biopsies, and their role has gotten progressively stronger in recent years. PMID- 17412154 TI - Idealism and realism. PMID- 17412155 TI - Deauthorization: the insidious new payer trick. PMID- 17412156 TI - Building virtual communities of practice. PMID- 17412157 TI - Seymour Ochsner. PMID- 17412158 TI - Metrics and NRDR: what is "near dear" and how will it affect radiology's future? PMID- 17412159 TI - What to do? PMID- 17412160 TI - What is wrong with 007? Lessons for leaders. PMID- 17412161 TI - Value-based competition in health care. PMID- 17412162 TI - The basics of restrictive covenants. PMID- 17412163 TI - The British National Health Service: crisis and transformation. PMID- 17412164 TI - Technology requirements for the optimal communication of critical test results. PMID- 17412165 TI - The deficit reduction act of 2005: policy, politics, and impact on radiologists. AB - The Deficit Reduction Act of 2005 (DRA) includes several provisions affecting Medicare physician payment policy that could have a significant impact on radiologists. Imaging and computer-assisted imaging services performed in physicians' offices will be subject to new payment caps starting in 2007, with reimbursement for the technical component limited to the lesser of what would be paid under the Hospital Outpatient Prospective Payment System or the Medicare fee schedule. It eliminates the scheduled reduction in fee-schedule payments for physicians' services for 2006, leaving payments at 2005 levels. It captures the savings accruing from a Medicare administrative decision cutting technical component reimbursement for imaging of contiguous body parts, rather than allowing the savings to be redirected to other areas of physician payment. The imaging payment cuts will save Medicare an estimated $2.8 billion over 5 years. The impact on individual physicians will vary substantially, depending on the extent of technical component and global billing, how much of their practices are devoted to Medicare, and the imaging modalities in which they specialize. Although the DRA cuts payments for certain imaging services, it does nothing to control utilization, which is the prime mover driving spending. PMID- 17412166 TI - ACCF/ACR/SCCT/SCMR/ASNC/NASCI/SCAI/SIR 2006 appropriateness criteria for cardiac computed tomography and cardiac magnetic resonance imaging. A report of the American College of Cardiology Foundation Quality Strategic Directions Committee Appropriateness Criteria Working Group. AB - Under the auspices of the American College of Cardiology Foundation (ACCF) together with key specialty and subspecialty societies, appropriateness reviews were conducted for 2 relatively new clinical cardiac imaging modalities, cardiac computed tomography (CCT) and cardiac magnetic resonance (CMR) imaging. The reviews assessed the risks and benefits of the imaging tests for several indications or clinical scenarios and scored them based on a scale of 1 to 9, where the upper range (7 to 9) implies that the test is generally acceptable and is a reasonable approach, and the lower range (1 to 3) implies that the test is generally not acceptable and is not a reasonable approach. The mid-range (4 to 6) indicates an uncertain clinical scenario. The indications for these reviews were drawn from common applications or anticipated uses, as few clinical practice guidelines currently exist for these techniques. These indications were reviewed by an independent group of clinicians and modified by the Working Group, and then panelists rated the indications based on the ACCF Methodology for Evaluating the Appropriateness of Cardiovascular Imaging, which blends scientific evidence and practice experience. A modified Delphi technique was used to obtain first and second round ratings of clinical indications after the panelists were provided with a set of literature reviews, evidence tables, and seminal references. The final ratings were evenly distributed among the 3 categories of appropriateness for both CCT and CMR. Use of tests for structure and function and for diagnosis in symptomatic, intermediate coronary artery disease (CAD) risk patients was deemed appropriate, while repeat testing and general screening uses were viewed less favorably. It is anticipated that these results will have a significant impact on physician decision making and performance, reimbursement policy, and future research directions. PMID- 17412167 TI - From medical invention to clinical practice: the reimbursement challenge facing new device procedures and technology--part 2: coverage. AB - This paper, the second of 3 that discuss the reimbursement challenges facing new medical device technology in various issues of this journal, explains the key aspects of coverage that affect the adoption of medical devices. The process Medicare uses to make coverage determinations has become more timely and open over the past several years, but it still lacks the predictability that product innovators prefer. The continued uncertainty surrounding evidence requirements undermines the predictability needed for optimal product planning and innovation. Recent steps taken by the Centers for Medicare and Medicaid Services to provide coverage in return for evidence development should provide patients with access to promising new technologies and procedures while generating important evidence concerning their effectiveness. PMID- 17412168 TI - Strategy development for anticipating and handling a disruptive technology. AB - The profession of radiology has greatly benefited from the introduction of new imaging technologies throughout its history. Therefore, it would seem reasonable for radiologists to believe that the emergence of a new imaging technology can generally be foreseen with sufficient advance notice to allow the appropriate levels of time, effort, and money to be devoted toward incorporating it into radiology practice. However, in his seminal work, Christiansen characterized a new form of technologic innovation, known as "disruptive technology," whose emergence often heralds the replacement of market leaders in an industry by competitors who are quicker in adopting and deploying the new technology. This article briefly describes the phenomenon of disruptive technology and addresses the challenges that organizations face in dealing with disruptive technology. The article raises 4 questions about the future of radiology: (1) Are health care and radiology vulnerable to disruptive technology? (2) What kinds of change may be in store for the radiology profession? (3) Can the radiology profession prepare itself to recognize and respond to a disruptive innovation among a group of new imaging technologies? and (4) How should a radiology organization decide whether to invest significant resources in a potentially disruptive technology? This article addresses these questions by reviewing key insights from leading "gurus" in the fields of competitive strategy and technology management and applying them to radiology. This illustrates how and why (despite past successes) the radiology profession may still have a blind spot in recognizing and handling disruptive technologies. PMID- 17412169 TI - Practice accreditation for radiation oncology: quality is reality. AB - The Radiation Oncology Practice Accreditation Program of the ACR is a voluntary program that was established in 1986. The processes of application and review are described. The most common causes of the deferral of accreditation are listed, as well as resources to help in preparing for an accreditation application. The costs of the survey and accreditation process are comparable with those of other accreditation programs outside of radiation oncology and the one competing radiation oncology accreditation program. The potential benefits of the program, financial and nonfinancial, are discussed, including the benefits to the specialty and to our patients. Possible integration with other programs, such as the pay-for-performance initiative and the maintenance of certification, are considered. PMID- 17412170 TI - Computer-based learning in medical education: a critical view. AB - A number of reports in the past decade have concluded that the medical education system must be reformed. These reports often call for the incorporation of new technologies into the educational process. One technology that has found its way into the medical educational environment is the personal computer. This essay critically examines the introduction of the personal computer into medical education, focusing on computer-based learning (CBL). The author concludes that evidence demonstrating the effectiveness of CBL is weak, and evidence supporting the notion that CBL enhances learning (compared with traditional methods) is weaker still. The author also argues that the decision to use CBL in the place of face-to-face traditional instruction has important negative implications for participants in medical education and may undermine the important role of patients in the education of doctors. PMID- 17412171 TI - Radiology order entry with decision support: initial clinical experience. AB - PURPOSE: To determine whether an appropriately designed computerized order entry system for radiology can be clinically accepted and influence ordering practices. MATERIALS AND METHODS: An intranet-based outpatient ordering and scheduling system was designed and implemented beginning in 2001. Indications used to request imaging have been standardized and keystrokes minimized by using menus. The system offers online scheduling and provides patient reminders, preparation instructions, and driving directions. Since November 2004, examination requests have been given utility scores on the basis of the indications provided. Comparative scores for other types of imaging examinations are displayed alongside the scores for the examinations requested. Physicians' performance is tracked, and senior clinicians counsel physicians with many low-scoring examinations. Data collected from the order entry system were used to evaluate rates of use, examinations with low "utility scores," and changes in the scores over the first year of use. RESULTS: The use of the order entry system has increased steadily, currently constituting 75% of all potential outpatient studies. Since the addition of decision support in November 2004, almost 72,000 examinations have been scored. The highest number of low utility examinations were imaging of the spine, either computed tomography or magnetic resonance imaging. The percentage of low utility examinations declined from 6% to 2% overall. The amount of the decline was greatest for primary care physicians and for those who interacted with the computer themselves rather than through office staff members. CONCLUSIONS: Computerized order entry with decision support can be widely accepted by clinicians and can have an impact on ordering practices. PMID- 17412172 TI - Impact of electronic resident evaluation systems on the timeliness, rate, and quality of responses. Survey of Association of Program Directors in Radiology members. AB - OBJECTIVE: To determine the impact of electronic resident evaluation systems on the timeliness, rate, and quality of responses. METHODS: Surveys were mailed electronically to the membership of the Association of Program Directors in Radiology, which included the directors of 158 residency programs. Respondents were instructed to send 1 response from each program. Information gathered included the use of electronic compared with paper-based evaluation systems and the overall level of satisfaction with such systems (rated on a 5-point scale with 1 = dissatisfied and 5 = satisfied). Questions were asked regarding the impact of electronic systems on the timeliness, rate, and quality of responses. RESULTS: Seventy-seven responses were received, for an estimated response rate of 49% on a per-program basis. Of these 77 respondents, 45 (58%) used electronic systems and 32 (42%) used paper-based systems. The median level of satisfaction was equivalent (4 = somewhat satisfied) for both groups. Of the 45 respondents using electronic systems, 26 (58%) reported increased response rates, 16 (36%) reported no change, and 3 (7%) reported decreased response rates compared with paper-based systems. Regarding the timeliness of responses, 31 (69%) reported faster response rates, 10 (22%) reported no change, and 4 (9%) reported slower rates compared with paper-based systems. Regarding the quality of responses, 25 (56%) reported no change, 12 (27%) reported improved quality, and 8 (18%) reported worse quality compared with paper-based systems. CONCLUSION: Electronic systems are generally associated with an improved response rate and enhanced timeliness of responses in comparison with paper-based systems, without adversely affecting the quality of responses. PMID- 17412173 TI - Physics education in medical imaging. PMID- 17412174 TI - Imaging for renal colic and hematuria. PMID- 17412175 TI - A Canadian on Capitol Hill. PMID- 17412176 TI - Henry Kuhnrath Pancoast. PMID- 17412177 TI - Lame ducks and legislators. PMID- 17412178 TI - Mother, may I ... PMID- 17412179 TI - Medical tort reform: a novel approach. PMID- 17412180 TI - Progress in patient safety: real or imagined? PMID- 17412181 TI - Cognitive therapy. PMID- 17412182 TI - ACR Appropriateness Criteria on suspected small-bowel obstruction. AB - With no one generally accepted approach to evaluate patients with suspected small bowel obstruction (SBO), standard CT has emerged as the preeminent imaging modality and should be considered in the initial evaluation of patients with suspected high-grade SBO. Playing less significant roles in the diagnosis of acute SBO are barium enemas and the small-bowel examination. PMID- 17412183 TI - From medical invention to clinical practice: the reimbursement challenge facing new device procedures and technology--part 3: payment. AB - This paper, the last of 3 that discuss the reimbursement challenges facing new medical device technology in various issues of this journal, addresses the structural diversity of Medicare's various payment systems. These systems vary widely in how they establish prices, how they incorporate new technologies and procedures, and the means by which they are updated and maintained. Their importance extends beyond Medicare because other payers use these payment rates as a basis for setting rates of their own. Device manufacturers and medical practitioners must often navigate several of these payment systems concurrently to ensure that technologies and procedures (that are already coded properly and covered) receive a fair payment rate. It is important to recognize that coverage can be undermined without adequate payment and that this situation will dampen further product innovation. The 3 papers, taken together, document the challenges posed by insurer reimbursement policies and show that a close working relationship between the manufacturers that develop new medical technologies and physician practitioners is needed if reimbursement hurdles are to be managed and medical innovation is to continue. PMID- 17412184 TI - Preauthorization of CT and MRI examinations: assessment of a managed care preauthorization program based on the ACR Appropriateness Criteria and the Royal College of Radiology guidelines. AB - PURPOSE: To evaluate computed tomography (CT) and magnetic resonance imaging (MRI) utilization patterns before and after the implementation of a preauthorization program based on the ACR Appropriateness Criteria((R)) and the guidelines of the Royal College of Radiologists. MATERIALS AND METHODS: All CT and MRI requests received at the preauthorization center and CT and MRI examinations actually performed were identified by our health care service's centralized computerized database between January 1, 2000, and December 31, 2003. The obligatory preauthorization of CT and MRI requests was established for CT in September 2001 and for MRI in February 2002. All ambulatory CT and MRI examination requests sent for approval during the study period by most of our health care physicians were included in the study. The preauthorization program model is presented, and multiple parameters were evaluated from January 2000 to December 2003, before and after preauthorization was established. RESULTS: Before preauthorization was required, the CT and MRI utilization rates were constantly increasing by 20% and 5% per year for CT and MRI, respectively. After preauthorization was implemented, CT and MRI annual performance rates decreased from 25.9 and 7 examinations per 1,000, respectively, in 2000 to 17.3 and 5.6 examinations per 1,000, respectively, in 2003. The decreases in the utilization of MRI and CT imaging between 2001 and 2003 were 9% (12,129 compared with 11,070 MRI examinations) and 33% (81,223 compared with 57,204 CT examinations), respectively, resulting in substantial, statistically significant cost savings. The deferral rate ranged from 7.5% to 12.2% (mean = 9.8%) for CT and 13.9% to 21.4% (mean = 17%) for MRI. Deferred cases in CT were most commonly in neuroradiology, musculoskeletal radiology, and CT angiography (ranges of deferred cases 9% to 12%, 11% to 12%, and 10% to 12%, respectively). Deferred cases in MRI were most commonly in abdominal and chest radiology (ranges of deferred cases 32% to 37% and 20% to 31%, respectively). Computed tomography was more commonly utilized inappropriately by pediatric professions, and MRI was more commonly utilized inappropriately by medical subspecialty professions. CONCLUSION: Preauthorization of CT and MRI requests results in a substantial decrease in utilization of these modalities with reduction in imaging costs. PMID- 17412185 TI - Emergency department abdominal computed tomography for nontraumatic abdominal pain: optimizing utilization. AB - OBJECTIVES: To identify predictors of positive computed tomographic (CT) yield and to measure the impact of CT yield on the disposition of patients referred for computed tomography after presenting to an emergency department with nontraumatic abdominal pain. MATERIALS AND METHODS: Computed tomographic reports, laboratory data, and emergency department and hospital records were retrospectively analyzed in 604 consecutive patients undergoing CT examinations. Computed tomographic yield was correlated to age, gender, leukocyte count, specified precomputed tomography clinical diagnosis, and patient disposition. RESULTS: Forty-eight percent of CT scans (298 of 621) had positive results. Computed tomographic results were positive in 76% of children (13 of 17) and 47% of adults (285 of 604) (P < .03) and in 45% of female patients (155 of 343) and 51% of male patients (143 of 278) (P < .2). Fifty-two percent of CT scans (223 of 426) with and 38% (75 of 195) without specified precomputed-tomography clinical diagnoses had positive results (P < .01). Fifty-eight percent of CT scans (161 of 278) with elevated and 40% of CT scans (135 of 336) with normal patient leukocyte counts had positive results (P < .001). Sixty-seven percent of patients (171 of 256) admitted and 35% of patients (127 of 365) discharged had positive CT results (P < .001). Computed tomography revealed unsuspected diagnoses in 27% of patients (165 of 621). Thirteen percent of patients (12 of 93) without any clinical predictors for positive CT yield were admitted after positive CT results. Thirty-eight percent of patients (104 of 273) with clinically suspected diagnoses requiring admission were discharged after negative CT results. CONCLUSION: Clinical indicators of positive CT yield include pediatric age, leukocytosis, and a specified precomputed-tomography diagnosis. Positive CT results are a predictor for hospital admission. In one quarter of cases, computed tomography identifies clinically unsuspected diagnoses and thereby adds information important for patient management, even after clinical evaluation. PMID- 17412186 TI - Shaping the future: maintenance of board certification and quality care. AB - This review is a summary of a presentation at the 2005 Intersociety Conference on training for the future of radiology. Certification by all 24 boards of the American Board of Medical Specialties, including the American Board of Radiology (ABR), has changed significantly. All primary and subspecialty certificates issued by the ABR are currently limited to 10 years, and diplomates of the board must maintain their certification by completing the ABR's maintenance of certification (MOC) program. The program consists of 4 components (professional standing, lifelong learning and self-assessment, cognitive expertise, and practice performance) and 6 competencies (medical knowledge, patient care, interpersonal skill, professionalism, practice-based learning and self improvement, and systems-based practice) that are the key elements to be incorporated within the concepts of continuous quality improvement. The result, over time, is intended to have a major impact in the quality of patient care in terms of outcomes and best practices. How adults learn in the environment of expanding knowledge and the electronic distribution of content is a substantive question requiring research, data, and change. We must seize the opportunity to explore adult learning and the process of MOC. Time-limited certification and MOC can become catalysts for future training requirements, for the design of training pathways, and for certification methodologies. Shaping the future is a noble task requiring leadership, vision, patience throughout change, and creativity. PMID- 17412187 TI - The effect of international teleradiology attending radiologist coverage on radiology residents' perceptions of night call. AB - PURPOSE: The purpose of this study was to determine the effects of international teleradiology attending radiologist coverage (ITARC) of emergency examinations on radiology residents' perceptions of night call. METHODS: A survey was administered at 2 different radiology residency programs that have attending radiologists who cover the night shift via teleradiology from Israel 5 nights per week. The survey consisted of 12 questions concerning residents' education and anxiety during on-call shifts and the effects of ITARC on these aspects of residency training. The questions were answered on a scale ranging from 1 to 5, with 3 being neutral. RESULTS: The radiology residents felt that ITARC improved the on-call learning experience (score = 3.7; 1 = much worse, 5 = much improved). The residents felt neutral about the statements "Review of cases with the attending radiologist over the telephone is comparable educationally to having the attending radiologist in person at the workstation" (score = 3.0) and "Having an attending radiologist easily available diminishes the need for me to commit to a diagnosis on my own and is therefore detrimental to my education" (score = 2.9; 1 = strongly disagree, 5 = strongly agree). The residents' stress levels on call were high without ITARC (score = 1.8; 1 = very high, 5 = very low) and moderate with ITARC (score = 2.7). The residents' anxiety levels before a night on call were moderate without ITARC (score = 2.9; 1 = very high, 5 = very low) and low with ITARC (score = 3.7). CONCLUSIONS: Radiology residents felt that ITARC improved their educational experience. International teleradiology attending radiologist coverage also decreased radiology residents' stress and anxiety related to on-call shifts. PMID- 17412188 TI - American College Of Radiology/Society of Breast Imaging curriculum for resident and fellow education in breast imaging. AB - The ACR and the Society of Breast Imaging have revised the curriculum for resident and fellow education in breast imaging on the basis of substantial changes in breast imaging practice since the initial curriculum was published in 2000. This curriculum provides guidance to academic chairs, residency program directors, and academic section chiefs in assessing and improving their residency and fellowship training programs and indicates to residents and breast imaging fellows the topics they need to learn and the experience they should try to acquire during their training. Radiologists already in practice also may find the curriculum useful in outlining the material they need to know to remain up to date in the practice of breast imaging. PMID- 17412189 TI - Qui tam means trouble. PMID- 17412190 TI - PACS: the long and winding road. PMID- 17412191 TI - Eugene P. Pendergrass. PMID- 17412192 TI - Development of a new instructional tool to increase the diagnostic accuracy of radiology residents interpreting emergency pediatric neuroradiology studies. PMID- 17412194 TI - Coding of ultrasound abdominal aortic aneurysms. PMID- 17412195 TI - The treasure before you. PMID- 17412196 TI - Quality begins at home. PMID- 17412197 TI - Women in radiation oncology: time to break through the glass ceiling. PMID- 17412198 TI - The need for the liberal arts. PMID- 17412199 TI - Wilbur Cohen and the origins of medicare. PMID- 17412200 TI - The stark reality about shared nuclear medicine imaging equipment leasing arrangements after January 1, 2007. PMID- 17412201 TI - How to think about insurance: the economics of risk and how it may affect our practice. AB - Health insurance has evolved, over time, due to changing public needs, changing supplier offerings, and advances in technology. Understanding the motivations for change and the effects on radiology practice can help our leadership respond appropriately. PMID- 17412202 TI - Buy-sell options for radiology: what works and why. AB - Buy-sell agreements for shareholders entering and leaving a radiology practice are different from those commonly used in other business endeavors. This paper explores the reasons for these differences, focusing on the culture of radiology and its unique influence on the buy-sell process. Buy-sell methodologies commonly used in most business transactions are described, and basic principles that influence these methodologies are discussed. The reasons these traditional methods are not applicable to most radiology groups are explored in depth. The paper concludes with a presentation of several workable buy-sell options for radiology practices. The strengths and weaknesses of these options are enumerated, so that each group can customize the option that best suits its needs. PMID- 17412203 TI - Facts and controversies about radiation exposure, part 1: controlling unnecessary radiation exposures. AB - In this 2-part article, the authors address the need to put in perspective the risks of radiation exposure in the rapidly changing field of radiology, considering the current state of knowledge of effects at low levels. The article is based on the content of the refresher course RC 516 presented at the Radiological Society of North America's 2005 annual meeting. After a brief review of epidemiologic studies, part 1 contains a discussion of typical radiation doses experienced in medicine, by both patients and professionals, and it concludes with a description of practical approaches to reduce unnecessary exposures. Part 2 of the article addresses a special concern for the unborn and discusses advisory and regulatory cancer risk estimates based mainly on epidemiologic studies. The limitations of epidemiologic studies at low-level exposures and recent new findings in radiobiology, some of which are summarized, challenge the notion that any amount of radiation causes adverse effects. PMID- 17412204 TI - Ethical conduct of radiology research with human participants. AB - Innovations in the field of radiology have been and continue to be possible through research that includes patients or healthy volunteers, or both, as research subjects. This article describes the principles that guide the ethical conduct of research as well as the procedures by which those principles are implemented, with a focus on how the principles are relevant to human research in radiology. Furthermore, we describe particular ethical challenges that radiology researchers are likely to confront. These include distinguishing research endeavors from innovative treatment, determining the acceptability of randomization and placebos, compensating for the "therapeutic misconception" of research subjects, and deciding when to disclose test results and incidental findings from research to subjects. We offer suggestions for anticipating and resolving such issues. PMID- 17412205 TI - Job interviewing skills for radiologists. AB - Physicians are interviewed for most residency and staff positions. This article addresses the skills and preparation helpful to do well on a radiology job interview. The topics covered in the article include research before an interview, practicing for the interview, and other advice for the interview day. PMID- 17412206 TI - Funding for graduate medical education. AB - The education of radiology residents and fellows is a vitally important but costly process. This paper reviews the most common methods of funding graduate medical education. The majority of graduate medical education in the United States is funded by Medicare, but there are caps on the number of trainees allowed, and the government is cutting payments. Academic medicine, particularly academic radiology, is at a point of crisis today if new methods to provide additional support are not found. PMID- 17412207 TI - Plain-radiographic image labeling: a process to improve clinical outcomes. AB - PURPOSE: To determine the rate of film-labeling errors and to describe a process for improved plain-film image labeling and the clinical outcomes from this process improvement. METHODS: Image-labeling errors (absent or incorrectly assigned left or right lateral identifier marker, absent or incorrect patient identifying number, absent or incorrect examination date, incorrect marker placement, absent technologist initial marker, or incorrect body-part order) were measured among 2,536 consecutive plain-film radiographs over a 2-week period. Following a process improvement initiative based on failure mode effectiveness analysis, left-side and right-side indicator markers, patient demographics, and date labels were identified as the most common sources of error. An improvement initiative using larger and colored left and right lateral indicator markers, an automated process to label patient demographics, and direct patient verification of identification was begun. The numbers of labeling errors were again assessed in 2,421 consecutive plain radiographs over a 2-week period. The error rates before and after the improvement initiatives were compared. RESULTS: Plain radiographic labeling errors occurred in 62 of 2,536 (2.4%) images before the improvement initiative. Labeling errors were reduced to 17 of 2,421 (0.70%; 95% exact binomial confidence interval, 0.4% to 1.1%; P < .001, chi-square test) by using the improvement tools. CONCLUSIONS: Plain radiographic image labeling can be improved using bar-code reading of patient demographic information, linked to patient Digital Imaging and Communications in Medicine modality work lists and image printing. Patient verification of demographic information is key and can be electronically managed. Lateral marker identification can be improved with larger (more easily read) and color-coded indicators. PMID- 17412208 TI - What you should know about handling digital studies generated outside your practice. PMID- 17412209 TI - What are the national radiation doses? PMID- 17412210 TI - When is imaging helpful for patients with back pain? PMID- 17412211 TI - Keep AFIP. PMID- 17412212 TI - Robert Reid Newell. PMID- 17412213 TI - Re: "Deauthorization: the insidious new payer trick". PMID- 17412215 TI - Maintenance of certification: tsunami ahead? PMID- 17412217 TI - Offshore radiology: the legal questions. PMID- 17412218 TI - The AFIP and the tragedy of the commons. PMID- 17412219 TI - 2006 ACR presidential oration. The ring in the radiograph: profession and principle. AB - A profession demands unconditional acceptance of the inalienable responsibilities to those it serves. Such obligations to patients, colleagues, and society infuse the specialty of radiology with meaning that elevates it to a profession and distinguishes radiologists' efforts from a simple occupation. The recognition and fulfillment of these duties are also how the ACR differentiates itself from an ordinary trade association. Professional insight provides a platform for analysis of and response to the issues confronting our specialty from the perspective of our founding principles and ethics. Judged by its history and recent leadership, the ACR has served and continues to serve this critical function by providing commitment, direction, and voice for our profession's core values. PMID- 17412220 TI - ACR Appropriateness Criteria on headache-child. AB - In a pediatric patient with an isolated headache or a classic migraine unaccompanied by neurologic signs, presence of a seizure, or supporting historical data, an imaging workup is usually not indicated. For a sudden severe headache or a headache with positive neurologic signs or symptoms or supporting historical data, MRI or CT should be considered. For the acute severe (thunderclap) headache, CTA, MRA, or catheter angiography may be appropriate. PMID- 17412221 TI - The 2006 ACR Forum: cardiovascular imaging: learning from the past, strategies for the future. AB - This paper summarizes the 2006 ACR Forum, which explored the history of the relationship between radiology and cardiovascular imaging and sought to explore strategies by which radiology could cope with similar challenges in the future. Key topics include: competition between radiology and other medical specialties, the importance of cardiac imaging, the relative merits of cardiologists and radiologists as cardiovascular imagers, and specific recommendations for radiology leaders in the areas of education, research, clinical practice, and policy. PMID- 17412222 TI - Facts and controversies about radiation exposure, part 2: low-level exposures and cancer risk. AB - In this 2-part article, the authors address the need to put in perspective the risks of radiation exposure in the rapidly changing field of radiology, considering the current state of knowledge of effects at low levels. The article is based on the content of the refresher course RC 516 presented at the Radiological Society of North America's 2005 annual meeting. In part 1, the authors presented a brief review of epidemiologic studies, a discussion of typical radiation doses experienced in medicine by both patients and professionals, and the description of practical approaches to reduce unnecessary exposures. Part 2 addresses a special concern for the unborn and discusses advisory and regulatory cancer risk estimates based mainly on epidemiologic studies. The limitations of epidemiologic studies at low-level exposures and recent new findings in radiobiology, some of which are summarized, challenge the notion that any amount of radiation causes adverse effects. PMID- 17412223 TI - Radiation oncology physicists will need to better understand medical imaging. AB - Imaging is affecting radiation oncology at a dramatically advancing pace and scale and is likely to create a transformation to individualized, biologically conformal radiation therapy. Deploying and improving imaging technologies and ensuring their correct uses in treatment planning and delivery are the responsibilities of radiation oncology physicists. The potential magnitude of errors arising from the incorrect use of imaging may be far greater than that resulting from typical errors in dose calibration. A major effort is required for radiation oncology physicists to raise the quality assurance of image guidance to a level comparable with that achieved in the maintenance of dosimetric performance. Most radiation oncology physicists lack adequate knowledge to assume this emerging responsibility. Their knowledge of imaging must be enhanced, in most cases through on-the-job training and self-learning. Effective learning strategies include routine interactions with diagnostic radiology and nuclear medicine physicists and physicians and the use of educational opportunities provided by professional organizations and vendors. PMID- 17412224 TI - Self-assessment for maintenance of certification. AB - Self-assessment is paired with continuing medical education in the "lifelong learning " component of the American Board of Radiology's (ABR) Maintenance of Certification program. The literature on the use of self-assessment in higher education and the professions is instructive with regard to the distinctiveness of this form of adult learning. This article reviews pertinent literature, summarizes the findings, and applies the principles to the creation and use of "self-assessment modules" (SAMs) for the maintenance of certification. Data on the use of ABR-qualified SAMs are reported. On completion of this article, the reader should be able to describe the nature, role, and best practices of self assessment; apply these concepts to the creation or use of a SAM; and identify future opportunities for SAM development and use that will enhance the lifelong learning of radiologists. PMID- 17412225 TI - Valuation of imaging centers: alternative methods and detailed description of the discounted cash flow approach. AB - Medical imaging centers are an increasingly integral part of the medical services landscape in America. There are many instances in which owners and potential buyers of these enterprises want to ascertain the value of the businesses. There is an industry of professionals who provide expert valuation services for many types of businesses using various recognized alternative methods, some of which are more appropriate than others when valuing an imaging center. The federal government has prescribed parameters for all valuations if they lead to transactions in which fair market value is mandated, and it also expects transactions to adhere to more generalized laws relating to entities that provide services to Medicare patients. Radiologists who own, or who are contemplating ownership of, imaging center operations need to understand the principles of valuation, specifically the factors that are involved in a discounted cash flow determination of fair market value. PMID- 17412226 TI - Use of computer databases to reduce radiograph reading errors. AB - Errors in reading radiologic images create health risks to patients and can adversely affect physicians through litigation and increased insurance costs. This study assessed the usefulness of a radiology teaching database for training physicians to improve their diagnostic accuracy. This tool is particularly important in pediatric radiology because children have unique anatomies and pathologies, and most physicians are generally more familiar with adults. The case collection consisted of normal cases and cases that had changes in the initial interpretations from an existing database of pediatric radiologic images at the Cincinnati Children's Hospital Medical Center. Physicians in the radiology department were surveyed regarding their opinions on the case collections. The most frequent users of the collection were residents and fellows, who used the collection primarily for learning and reference. Attending radiologists used the collection for teaching and daily film reading. The most commonly used cases involved head computed tomography, brain magnetic resonance, and skull radiography. All respondents recognized the value of the database for the enhancement of residents' and fellows' training. Residents and fellows expressed the desire to add complete abnormal and frequently missed case collections to the normal collection. Physicians who used the collections did so very frequently, but many experienced physicians never accessed the collection. Inexperienced physicians' reliance on reference books compared with Internet resources highlights a need for computer-based teaching and reference tools. This study illustrates that easy-to-use computer-based teaching files are useful for training physicians and as reference tools for experienced physicians. PMID- 17412227 TI - New year, new codes...and all new confusion. PMID- 17412228 TI - Walls and bridges. PMID- 17412230 TI - Harold G. Jacobson. PMID- 17412231 TI - Re: "Ultrasound: is there a future in diagnostic imaging?". PMID- 17412233 TI - Focusing on the big picture. PMID- 17412234 TI - CT screening for lung cancer: appearances can be deceiving. PMID- 17412235 TI - Reliably better, faster, and cheaper souffles... PMID- 17412236 TI - Patient-focused radiology: the value of customer service? PMID- 17412237 TI - The new house of blues. PMID- 17412238 TI - What you need to know before purchasing a PACS. PMID- 17412239 TI - The economic environment of 2006: how safe is your practice and what you can do to protect it. AB - In 2006, there was no doubt that imaging was in the crosshairs of just about everyone who had a role in legislation, regulation, and payment involving these medical services. The information in this article was presented at the ACR's Annual Meeting and Chapter Leadership Conference in Washington, DC, on May 25, 2006. It describes the factors contributing to the current economic environment for medical imaging, the external responses to those factors, the dangers to practicing radiologists, and potential strategies to mitigate the damage. PMID- 17412240 TI - HOPPS: evolution of a CMS process. AB - Medicare's hospital outpatient prospective payment system (HOPPS) was initially developed in response to the rapid rise in Medicare's outpatient expenses between 1980 and 1991. The Balanced Budget Act of 1997 mandated HOPPS, with an implementation date of August 1, 2000. Unlike the Medicare Physician Fee Schedule, the Centers for Medicare and Medicaid Services (CMS) used hospital charge data to develop the ambulatory patient classification (APC) payment weights. During its evolution as a payment system, Congress mandated the creation of an advisory panel as well as the removal of diagnostic mammography from the APCs. The Deficit Reduction Act proposes applying HOPPS for paying technical fees in nonhospital settings. PMID- 17412241 TI - Valuing the professional work of diagnostic radiologic services. AB - The challenges to the technical component (TC) of radiology services as a result of the Deficit Reduction Act of 2005 have been widely publicized, but a number of regulatory changes will affect the professional component (PC) of our physician work. The third 5-year review is complete, and although there were no major reductions in work relative value units (RVUs) for radiology, proposed budget neutrality adjustments to physician work RVUs will potentially reduce payments for both the PC and global payments. Additionally, if there is no congressional intervention, reductions in the conversion factor will further decrease Medicare payments. In the article, the history of the radiology relative value scale and the methodology of American Medical Association/Specialty Society Relative Value Scale Update Committee's valuation of physician work are reviewed. The results of the third 5-year review are presented. Future challenges to radiology physician work valuation are discussed, including outsourcing, the Medicare Payment Advisory Commission's search for overvalued services, and the bundling of physician services. Whereas the TC is compensation for performing a diagnostic test, ultimately, it is our physician work that defines our specialty, and challenges to our physician work will be vigorously defended by the ACR without compromise. PMID- 17412242 TI - The ACR Managed Care Committee: focusing private payers on radiology issues. AB - The ACR Managed Care/Private Payer Relations Committee is an important committee of the ACR Commission on Economics. This report reviews the committee's mission, structure, and processes and some of its current recommendations to ACR membership. The development of and participation in radiology advisory committees is a vital strategy in this process. Separating professional and technical charges, rather than submitting global charges, will help preserve radiology's professional integrity in the future. The Imaging Provider Report Card (IPRC) will allow radiology to define practice quality and performance in an era of pay for-performance reimbursement. The IPRC also provides an external blueprint on what each practice needs to do to improve itself. American College of Radiology accreditation plays a key role in certifying radiology's quality to both payers and patients. Sound business management, group governance, and business size are also important elements of professional practice success. Working together through the ACR promotes the integrity of our profession and the quality of care patients want and deserve. PMID- 17412243 TI - Advanced marketing: how to protect and advance your practice. AB - Radiology practices in the United States are facing mounting competitive challenges in many metropolitan areas. This competition is multifaceted and includes other traditional diagnostic imagers. However, the most severe pressures are from new entrants, such as primary and specialty physicians outside of radiology. The business science of marketing provides a set of solutions to help you fight back against this onslaught. In particular, advanced marketing techniques offer compelling strengths for raising awareness of the unique nature of our specialty among both the lay population and primary referrers. Applications of advanced marketing can be used to simultaneously hold and expand your turf. Relying on smarts rather than dollars, they also help overcome the fears that many radiology practices have about the high costs of traditional broadcasting. PMID- 17412244 TI - Recent trends in utilization of vascular ultrasound among radiologists, surgeons, cardiologists, and other physicians. AB - PURPOSE: To determine how much vascular ultrasound is performed by the various medical specialties and to assess the utilization trends among them in recent years. METHODS AND MATERIALS: The nationwide Medicare Part B databases for 1998 through 2004 were used. The authors studied the 15 Current Procedural Terminology, Version 4, codes in the 93000 series pertaining to vascular ultrasound and determined the volume of examinations performed under global and professional component claims. Using Medicare's physician specialty codes, utilization rates per 1,000 Medicare beneficiaries and trends in these rates were determined for radiologists, surgeons, cardiologists, other physicians, and independent diagnostic testing facilities (IDTFs) over the 6-year period. Market shares were calculated for the different specialties. RESULTS: In 2004, among all specialties, 171.8 vascular ultrasound studies were performed per 1,000 Medicare beneficiaries, an increase of 49% over 1998. For the various specialty groups, the 2004 utilization rates and percentage changes since 1998 were as follows: radiologists, 70.8 per 1,000 (+44%); surgeons, 44.6 per 1,000 (+47%); cardiologists, 23.1 per 1,000 (+93%); other physicians, 24.0 per 1,000 (+52%); and IDTFs, 9.3 per 1,000 (+12%). Market shares in 2004 were as follows: radiologists, 41.2%; surgeons, 26.0%; cardiologists, 13.4%; other physicians, 14.0%; and IDTFs, 5.4%. Although the percentage change in utilization rates over the 6 years was highest among cardiologists, the addition of new case volume was highest among radiologists. Radiologists added 1,110,961 new studies between 1998 and 2004, whereas surgeons added 713,251, cardiologists added 489,958, other physicians added 401,601, and IDTFs added 89,077. CONCLUSIONS: Radiologists have the largest market share in vascular ultrasound. Between 1998 and 2004, they added more new volume than other specialties. Utilization trends suggest that radiologists will continue to have a strong role in this major aspect of vascular imaging. PMID- 17412245 TI - How much software can you deploy in one sitting? PMID- 17412246 TI - Federal regulations (effective June 2006) require dose monitors on all new fluoroscopes: how will this help clinicians keep track of patient dose? PMID- 17412247 TI - Magnetic resonance cholangiopancreatography. PMID- 17412248 TI - Robert Stone. PMID- 17412249 TI - Quality--full steam ahead. PMID- 17412250 TI - Everyone is working harder. PMID- 17412251 TI - Quality improvement in radiology: white paper report of the 2006 Sun Valley Group meeting. PMID- 17412252 TI - Academic radiology: time to think outside of the box. PMID- 17412253 TI - What I learned as a patient. PMID- 17412254 TI - ACR Appropriateness Criteria on solitary pulmonary nodule. AB - The solitary pulmonary nodule is traditionally defined as a relatively spherical opacity 3 cm or less in diameter surrounded by lung parenchyma. The choice of imaging test to evaluate solitary nodules is extensive. However, only 2 findings are considered to be sufficient to preclude further evaluation: calcification in a benign pattern and stability in size for more than 2 years. PMID- 17412255 TI - Quality--a radiology imperative: report of the 2006 Intersociety Conference. AB - Recent publications by the Institute of Medicine have pointed out the many medical errors that occur and the adverse effect of those errors on patient outcomes. These errors are often the result of problems within our health care systems. In an effort to improve patient care outcomes, many health care providers have developed improved policies and practices. Third-party payers are encouraging participation in quality efforts through a variety of mechanisms, including pay for performance, pay for participation, and the creation of centers of excellence. If the quality of health care is to be improved and monitored, appropriate metrics must be developed. Such metrics must be within the control of providers, measurable, and likely to improve patient outcomes. The participants of the 2006 Intersociety Conference developed 49 metrics, which are offered for adoption by health care networks, payers, and regulation agencies. PMID- 17412256 TI - Quality--a radiology imperative: interpretation accuracy and pertinence. AB - Physicians as a group have neither consistently defined nor systematically measured the quality of medical practice. To referring clinicians and patients, a good radiologist is one who is accessible, recommends appropriate imaging studies, and provides timely consultation and reports with high interpretation accuracy. For determining the interpretation accuracy of cases with pathologic or surgical proof, the author proposes tracking data on positive predictive value, disease detection rates, and abnormal interpretation rates for individual radiologists. For imaging studies with no pathologic proof or adequate clinical follow-up, the author proposes measuring the concordance and discordance of the interpretations within a peer group. The monitoring of interpretation accuracy can be achieved through periodic imaging, pathologic correlation, regular peer review of randomly selected cases, or subscription to the ACR's RADPEER system. Challenges facing the implementation of an effective peer-review system include physician time, subjectivity in assessing discordant interpretations, lengthy and equivocal interpretations, and the potential misassignment of false-positive interpretations. PMID- 17412257 TI - Determinants of radiologists' desired workloads. AB - PURPOSE: To examine what factors lead radiologists to desire different work hours. MATERIALS AND METHODS: Particular attention was paid to the intensity of work, measured in procedures per work hour, a potentially important variable previously largely ignored. Weighted data from the ACR's 2003 survey of radiologists were used. Multiple linear regression was used to analyze the influences of characteristics of individual radiologists and the practices in which they worked on their desired annual workloads. RESULTS: Increasing the work intensity of the average radiologist by 1 procedure per hour (11%) decreased the annual desired work hours by 2.3 hours (0.1%). Older radiologists sought smaller workloads. For example, the desired annual work hours of radiologists aged 65 years and older were 262 annual hours fewer than those of radiologists aged 45 to 54 years. The desired annual work hours of radiologists in academic practices were more (by 213 hours) than those of radiologists in private, nonacademic radiology groups. In addition, the desired annual work hours of radiologists in practices situated in the West and Northwest census regions were fewer than those of radiologists in practices in the South. CONCLUSION: Radiologists who worked more intensively sought fewer annual work hours, but the effect of greater work intensity on desired hours was very small. The very small size of this effect makes it likely that radiology practices will continue, as they have in the past, to accommodate the increasing imaging workload in the United States by increasing the intensity of their work rather than by lengthening hours. PMID- 17412258 TI - The outpatient availability score: an alternative approach to measuring demand. AB - Radiology business managers are challenged daily to maximize access, improve utilization, and enhance the efficiency of very costly resources. They rely on a variety of reports, metrics, and indicators to make operational decisions. Among the many metrics managers track today, one of the more inexact is the determination of outpatient appointment availability. The outpatient availability score (OAS) was developed to improve how access to resources is predicted in the department of radiology at Massachusetts General Hospital by accounting for patient preference. The OAS uses a range of predictors (thresholds) to determine the low, medium, and high likelihood that patients will find appointments suitable for them. The OAS can be customized to fit individual operations by adjusting these thresholds as fluctuations in demand dictate. The OAS is more useful than other current methodologies for measuring availability, such as next appointment availability or third next appointment availability, for the following reasons: (1) the OAS forecasts availability for an extended period of time, (2) the OAS forecasts the quantity of available appointments, and (3) the OAS is a better indication of the department's ability to satisfy patients' appointment needs. The OAS used in conjunction with other measures (such as the third next available appointment) of availability affords managers a clearer picture of access and the ability to deal proactively with demand to run more efficient operations. PMID- 17412259 TI - How to transition to digital mammography. AB - To date, the transition to digital mammography has been cumbersome and difficult. In part, this is because digital mammography equipment could not be easily integrated into preexisting picture archiving and communication systems (PACS). Until the results of the ACR Imaging Network Digital Mammographic Imaging Screening Trial showed that digital mammography is more accurate than screen-film mammography for some patients, users have been reticent to switch. To make the transition as seamless as possible, several considerations should be made. First, an evaluation of a facility's current screen-film situation, including the condition of the analog units, work flow, patient capacity, staffing, and existing PACS components helps in understanding current status more clearly. After that, the facility's goals for a digital department should be determined. Next, the components of the digital imaging chain, the network infrastructure, and the PACS, as well as what clinical aspects each part of the chain affects, must be understood to avoid unexpected frustrations after the conversion is made. Then, education is required on the different acquisition options, display options, printing options, and archival options to facilitate the choice of equipment that will meet the digital goals set forth previously. Finally, a pace of conversion that is best suited to the facility should be determined. PMID- 17412260 TI - Turf wars in radiology: the future of radiology depends on research--and on your support of it! AB - The private practice radiology community relies on academic radiology departments to (1) do research and make advances in imaging science, (2) justify reimbursement for those advances, and (3) train new generations of radiologists. This has been largely a one-sided arrangement, with the academic departments footing the bill and the private practice community deriving the benefits and pretty much getting a free ride. But academic radiology is facing a crisis, and the research productivity of our specialty is in jeopardy. Leadership in research is a crucial element in the effort to keep out untrained and inexperienced physicians from other specialties who are largely motivated by the desire for profit. Better support from private practice radiologists is badly needed to help their academic colleagues maintain that research leadership. PMID- 17412261 TI - Medically unbelievable edits: unbelievable indeed. PMID- 17412262 TI - The virtues of optimism. PMID- 17412263 TI - Technical aspects of respiration-correlated 4-D CT for radiation therapy. PMID- 17412264 TI - Lewis Gregory Cole. PMID- 17412265 TI - Opening new educational frontiers. PMID- 17412266 TI - Appropriateness. PMID- 17412267 TI - Self-dealing in medical imaging: call for action. PMID- 17412268 TI - Saving radiology: a modest proposal for branding and promoting our specialty in 2007. PMID- 17412269 TI - Consumer-directed health plans: at last, data. PMID- 17412270 TI - Compliance programs: serious business for health care providers. PMID- 17412271 TI - Defining quality in radiology. AB - The introduction of pay for performance in medicine represents an opportunity for radiologists to define quality in radiology. Radiology quality can be defined on the basis of the production model that currently drives reimbursement, codifying the role of radiologists as being limited to the production of timely and accurate radiology reports produced in conditions of maximum patient safety and communicated in a timely manner. Alternately, quality in radiology can also encompass the professional role of radiologists as diagnostic imaging specialists responsible for the appropriate use, selection, interpretation, and application of imaging. Although potentially challenging to implement, the professional model for radiology quality is a comprehensive assessment of the ways in which radiologists add value to patient care. This essay is a discussion of the definition of radiology quality and the implications of that definition. PMID- 17412272 TI - What has happened to cardiac MRI? AB - Cardiac magnetic resonance imaging (MRI) has made significant technical advances over the past 20 years. There has been a steady increase in the number of scientific publications. Recent trends show that the rate of publications is increasing more rapidly for cardiology departments than for radiology departments. In examining PubMed citations, the majority of publications continue to be authored by both cardiologists and radiologists. Despite the growing research interest, relatively few clinically indicated cardiac MRI examinations are performed compared with competing tests. Competing technology and relatively poor reimbursement may in part explain the slow acceptance of cardiac MRI. PMID- 17412274 TI - Top 10 tips for a radiology medical-legal expert. AB - These 10 tips for a radiology medical-legal expert will improve your understanding of the unfamiliar maze of civil court rules, regulations, and opposing attorney tactics to make you a more effective expert witness. PMID- 17412275 TI - Indian radiologists in the United States: hierarchical distribution and representation. AB - PURPOSE: To evaluate both the distribution of radiologists of Indian ancestry and nativity in the United States in training and beyond and their representation in hierarchal positions in major radiology organizations. MATERIALS AND METHODS: The annual membership lists of the Radiological Society of North America (RSNA) and the ACR over the past decade were analyzed for the number and locations of radiologists of Indian ancestry or nativity. To evaluate the progression and current extent of enrollment of members of this cohort in positions of leadership in radiology organizations, their overall membership and their specific participation in RSNA and ACR committees were tabulated. Their listing in the latest directories of chairpersons and program directors in radiology was assessed. The distribution of ethnic Indian radiologists in training was calculated by examining resident rosters by academic program and state. RESULTS: Since the passage of the Immigration and Nationality Act of 1965, individuals of Indian ancestry have constituted a growing minority of radiologists. In 2002, they constituted 5.5% of all RSNA members. Recently, their numbers in training programs have increased greatly. They are now estimated to constitute 13.3% of all residents, predominantly because of the entry of the American-born offspring of Indian immigrants. Yet ethnic Indians are markedly underrepresented in the hierarchy of major radiology organizations. In both 1999 and 2003, there were no Indian-surnamed representatives on most major committees and none in board-level positions of both the ACR and the RSNA. The number of radiologists of Indian birth or background who are program directors has increased slightly over the past 10 years from 2:196 (1%) in 1993 to 7:196 (3.5%) in 2003. The representation of radiologists of Indian ethnicity on the mastheads of both the American Journal of Roentgenology and Radiology remains meager; fewer than 2% of these journals' assistant editors and editors are of Indian ethnicity. CONCLUSION: The percentage of radiologists identifiable as Indian by nativity or ethnicity is growing rapidly, predominantly because of an increase in the number of American-born trainees. Yet only a few more senior members of this group have secured positions of leadership in the specialty, restricting the number of ethnically similar role models for Indian junior members of the specialty to emulate. PMID- 17412276 TI - Successful recruiting of radiologists. AB - Hospital systems and radiology organizations often find it difficult to recruit new high-quality radiologists. This article discusses long-range planning for finding new radiologists, methods used to recruit prospective new members for a radiology department or group, how to carry out a satisfactory interview, and questions that are illegal to pose during an interview. The related topics of the state of the radiology job market and ways to reduce staff turnover are also addressed briefly. PMID- 17412277 TI - How the radiologic and nuclear medical communities can improve nuclear security. AB - Highly enriched uranium (HEU) is used to manufacture technetium-99m, the most widely used medical radioisotope in the world. Highly enriched uranium is also used to make nuclear bombs; 50 kg of HEU is enough to make a Hiroshima-type bomb. It is generally agreed that this technology is within the reach of a terrorist group; the main obstacle is acquiring HEU. Currently, as a legacy of the US and Soviet Atoms for Peace Program, there are civilian users of HEU in 40 countries, and about 1,000 kg are still being shipped each year. Unfortunately, the major international manufacturers of technetium-99m have been refusing to convert their production facilities to use low-enriched uranium (LEU), which cannot be used to make a nuclear bomb. Only 1% to 2% of the HEU is consumed in the process of producing technetium-99m. The remainder is accumulating in radioactive waste storage facilities. The radiologic and nuclear medical communities could make a tremendous contribution to a safer world by supporting the replacement of HEU with LEU in the production of technetium-99m. Low-enriched uranium is just as cost effective as HEU for the manufacture of technetium-99m and does not contribute to the risk for nuclear terrorism. PMID- 17412278 TI - What you can do to minimize system downtime. PMID- 17412279 TI - An update on the new curriculum for educating radiologists about physics. PMID- 17412280 TI - FDG-PET CT for tumor imaging. PMID- 17412281 TI - Dale Trout. PMID- 17412282 TI - Complement components, regulators and receptors are produced by human monocyte derived dendritic cells. AB - Complement and dendritic cells (DCs) are essential components of innate immunity. Both participate in local inflammation and moreover have roles in the initiation of the acquired immunity response and in the maintenance of tolerance. Recent studies have demonstrated the ability of DCs to synthesize C1q, C3, Factor I, Factor B and complement receptors 3 and 4. In this study, we demonstrate that human DCs are a source of other soluble complement proteins including C1q, C4b binding protein (C4BP), C7 and C8. Complement receptors (CR)1 and the CD18 chain (common for CR3 and CR4) were also present on DCs while CR2 was not detected. PMID- 17412283 TI - Expression of programmed-death receptor ligands 1 and 2 may contribute to the poor stimulatory potential of murine immature dendritic cells. AB - Recent data have revealed that Ag presentation by immature dendritic cells (imDCs) plays a role in establishing and maintaining T-cell tolerance, but the mechanism remains unclear. PD-L1 and PD-L2, ligands for programmed-death receptor 1 (PD-1), members of the expanding B7 family, were highlighted for their inhibitory role in T-cell responses. Here, we show that blockade of PD-1 ligands on imDCs resulted in enhanced T-cell proliferation, which is perhaps due to the enhancement of IL-2 production from DC-stimulated T cells. PD-1 ligands blockade on mDCs did not show a significant stimulatory effect as markedly as imDCs. The inhibitory effects of PD-1 ligands would be dependent on maturation status of DCs, where attenuated positive costimulatory molecules provided the opportunity for PD-1 ligands to exert their strong capacity. Our data are consistent with the hypothesis that imDCs have an inhibitory bias, and indicate that PD-L1 and PD-L2 contribute to the poor stimulatory capacity of imDCs. PMID- 17412284 TI - Ex vivo characterization of human thymic dendritic cell subsets. AB - Interactions between thymic dendritic cells (DC) and thymocytes are critical for proper development of T-cells. We identified human thymic DC populations on the basis of CD123, CD11c and CD14 expression. High levels of CD123 (IL-3R) and CD45RA defined the plasmacytoid DC (pDC) subset. Human thymic CD11c(+) DC expressed CD45RO and myeloid-related markers (CD13, CD33 and CD11b). CD11c(+) DC could be separated into two main subsets based on differential expression of CD14: CD11c(+) CD14(-) and CD11c(+) CD14(+) cells. Spontaneous production of IL 10 and IFNgamma without exogenous stimulation, was observed in the three DC subsets. Important phenotype modifications were observed in pDC cultures supplemented with IL-3. A down-regulation of CD123 and appearance of myeloid markers such as CD11b and CD11c on CD45RA(+) cells was noticed within the first 48h; at a later time there was a shift from CD45RA to CD45RO expression, as well as appearance of CD14 expression. CD11c(+) cells emerging in pDC culture did not express high levels of HLA-DR, CD83 and co-stimulatory molecules. This suggests an in vitro evolution of human thymic pDC toward a myeloid phenotype found in the CD11c(+) subset of thymic DC. PMID- 17412285 TI - Immunomodulating effect of inositol hexaphosphate against Aeromonas hydrophila endotoxin. AB - The present study was carried out to evaluate the effect of inositol hexaphosphate (IP6) administration on endotoxemia as an example of the systemic inflammatory response. Mice were divided into three groups as follows: First group, remained as a naive group injected intraperitoneally (i.p.) with PBS (pH 7.4; 0.2 ml/mice) at intervals parallel to the treated groups. The second group was injected i.p. with the lipopolysaccharide (LPS) of Aeromonas hydrophila once a week for four weeks at a dose of LPS suspension: 20 mg/kg mice/week. The third group was injected with the same LPS dose and synergistically intubated with IP6 three times a week for four weeks at a total dose of 4 0mg/kg. At different experimental periods (1, 2, 3 and 4 weeks), six animals from each group were sacrificed under mild diethyl ether anesthesia. Blood and sera were taken for the estimation of phagocytic activity, electrophoretic pattern of proteins and immunoglobulin levels. Also, a slice of liver was homogenized to estimate the respiratory burst enzymes activities and nitric acid synthesis. Histopathological changes of hepatic tissues were investigated. In the LPS-treated group, marked increase in the phagocytic activities and nitric oxide synthesis, and a decrease in hepatocyte catalase, total peroxidase and superoxide dismutase activities were observed. The histopathological features revealed a degeneration and highly mitotic division within the hepatic nuclei in addition to some karyomegaly and nuclear pyknosis. During the treatment period, liver sections of the LPS+IP6 group showed somewhat regenerative features. Reduction in the toxicity of free radicals by IP6 was observed and the IP6 effect seemed to be responsible for the observed ameliorative influence. PMID- 17412286 TI - Anti-beta 2-glycoprotein I antibodies of IgM class are linked to thrombotic disorders in young women without autoimmune disease. AB - Antiphospholipid autoantibodies particularly antibodies against beta2 glycoprotein I (anti-beta2GPI) are casually associated with thromboses in patients with autoimmune diseases. However, their exact prevalence and role in the pathogenesis of thromboses in the absence of autoimmune disease is still inconclusive. They might be particularly important when other risk factors of thrombosis are absent. We investigated antiphospholipid antibodies in 68 young women (aged <45yr at onset of the event, without autoimmune disease and with an otherwise low risk of thrombosis) in the stable period following myocardial infarction (MI), lacunar cerebral infarction (LACI) or deep vein thrombosis (DVT) and in 37 healthy age-matched controls. Patients had increased IgM anti-beta2GPI compared to controls (36.0, 11.5-49.5 vs. 17.50, 3.50-30.0 arbitrary units (AU), p<0.001), whereas no difference was obtained in other measured antibodies (anticardiolipin and antiphosphatidylserine (aPS) antibodies of IgG and IgM). IgM anti-beta2GPI positively correlated with some markers of increased coagulation potential and negatively with BMI (r=-039, p<0.005) and other parameters of the metabolic syndrome. In conclusion, we found that levels of IgM anti-beta2GPI are increased in young women suffering arterial or venous thromboses in the absence of other known autoimmune diseases and also in the absence of pronounced classical risk factors. We found that IgM anti-beta2GPI positively correlated with some markers of increased coagulation potential and negatively with parameters of the metabolic syndrome. Thus, it appears that elevated levels of IgM anti-beta2GPI are linked to thrombotic disorders in young women (without autoimmune disease) particularly when classical risk factors or the metabolic syndrome are absent. PMID- 17412287 TI - The SARS coronavirus spike glycoprotein is selectively recognized by lung surfactant protein D and activates macrophages. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) infects host cells with its surface glycosylated spike-protein (S-protein). Here we expressed the SARS-CoV S-protein to investigate its interactions with innate immune mechanisms in the lung. The purified S-protein was detected as a 210 kDa glycosylated protein. It was not secreted in the presence of tunicamycin and was detected as a 130 kDa protein in the cell lysate. The purified S-protein bound to Vero but not 293T cells and was itself recognized by lung surfactant protein D (SP-D), a collectin found in the lung alveoli. The binding required Ca(2+) and was inhibited by maltose. The serum collectin, mannan-binding lectin (MBL), exhibited no detectable binding to the purified S-protein. S-protein binds and activates macrophages but not dendritic cells (DCs). It suggests that SARS-CoV interacts with innate immune mechanisms in the lung through its S-protein and regulates pulmonary inflammation. PMID- 17412288 TI - Nutrition and immune system: certain fatty acids differently modify membrane composition and consequently kinetics of KV1.3 channels of human peripheral lymphocytes. AB - Potassium (K(+)) channels of human peripheral lymphocytes play a considerable role in the signalling processes required for immune responses. Modification of the fatty acid composition of the membrane influences the functions of various membrane enzymes and ion channels. We set out to establish how the incorporation of fatty acids with different carbon chain lengths and degrees of unsaturation into the cell membrane influences the function of K(V)1.3 channels of lymphocytes, thereby potentially modifying the immune responses of the cells. The incorporation of the fatty acids into the cell membrane was monitored by gas chromatography. Whole-cell patch-clamp experiments demonstrated that the polyunsaturated linoleic acid, arachidonic acid and docosahexaenoic acid all decreased the activation and inactivation time constants of the K(V)1.3 channels, but did not affect the voltage-dependence of steady-state activation and steady state inactivation of the channels. Treatment with the saturated palmitic acid, stearic acid and the monounsaturated oleic acid did not result in significant changes in the biophysical parameters of K(V)1.3 gating studied. We conclude that the incorporation of fatty acids unsaturated to different degrees into the cell membrane of lymphocytes influenced the rate of gating transitions but not the equilibrium distribution of the channels between different states. This effect depended on the degree of unsaturation and the chain length of the fatty acids: no effects of saturated and monounsaturated fatty acids (16:0, 18:0 and 18:1) were observed whereas treatment with polyunsaturated fatty acids (18:2, 20:4 and 22:6) resulted in significant changes in the channel kinetics. PMID- 17412289 TI - Photosynthesis research on yellowtops: macroevolution in progress. AB - The vast majority of angiosperms, including most of the agronomically important crop plants (wheat, etc.), assimilate CO2 through the inefficient C3 pathway of photosynthesis. Under ambient conditions these organisms loose about 1/3 of fixed carbon via photorespiration, an energetically wasteful process. Plants with C4 photosynthesis (such as maize) eliminate photorespiration via a biochemical CO2 pump and thus have a larger rate of carbon gain. The genus Flaveria (yellowtops, Asteraceae) contains not only C3 and C4 species, but also many C3-C4 intermediates, which have been interpreted as evolving from C3 to fully expressed C4 metabolism. However, the evolutionary significance of C3-C4Flaveria intermediates has long been a matter of debate. A well-resolved phylogeny of nearly all Flaveria species has recently been published. Here, we review pertinent background information and combine this novel phylogeny with physiological data. We conclude that the Flaveria species complex provides a robust model system for the study of the transition from C3 to C4 photosynthesis, which is arguably a macroevolutionary event. We conclude with comments relevant to the current Intelligent Design debate. PMID- 17412290 TI - Geometric robustness theory and biological networks. AB - We provide a geometric framework for investigating the robustness of information flows over biological networks. We use information measures to quantify the impact of knockout perturbations on simple networks. Robustness has two components, a measure of the causal contribution of a node or nodes, and a measure of the change or exclusion dependence, of the network following node removal. Causality is measured as statistical contribution of a node to network function, whereas exclusion dependence measures a distance between unperturbed network and reconfigured network function. We explore the role that redundancy plays in increasing robustness, and how redundacy can be exploited through error correcting codes implemented by networks. We provide examples of the robustness measure when applied to familiar boolean functions such as the AND, OR and XOR functions. We discuss the relationship between robustness measures and related measures of complexity and how robustness always implies a minimal level of complexity. PMID- 17412291 TI - The role of intragenomic recombination rate in the evolution of population's genetic pool. AB - In a simple computer model of population evolution, we have shown that frequency of recombination between haplotypes during the gamete production influences the effectiveness of the reproduction strategy. High recombination rates keeps the fraction of defective alleles low while low recombination rate or uneven distributed recombination spots change the strategy of genomes' evolution and result in the accumulation of heterozygous loci in the genomes. Even short fragment of chromosome with restricted recombination influences the genetic structure of neighboring regions. PMID- 17412292 TI - A new approach for estimating the efficiencies of the nucleotide substitution models. AB - In this article, a new approach is presented for estimating the efficiencies of the nucleotide substitution models in a four-taxon case and then this approach is used to estimate the relative efficiencies of six substitution models under a wide variety of conditions. In this approach, efficiencies of the models are estimated by using a simple probability distribution theory. To assess the accuracy of the new approach, efficiencies of the models are also estimated by using the direct estimation method. Simulation results from the direct estimation method confirmed that the new approach is highly accurate. The success of the new approach opens a unique opportunity to develop analytical methods for estimating the relative efficiencies of the substitution models in a straightforward way. PMID- 17412293 TI - The risk of extinction - the mutational meltdown or the overpopulation. AB - The phase diagrams survival-extinction for the Penna model with parameters: (mutations rate)-(birth rate), (mutation rate)-(harmful mutations threshold), (harmful mutation threshold)-(minimal reproduction age) are presented. The extinction phase may be caused by either mutational meltdown or overpopulation. When the Verhulst factor is responsible for removing only newly born babies and does not act on adults the overpopulation is avoided and only genetic factors may lead to species extinction. PMID- 17412294 TI - Evolution and development: past, present, and future. AB - The paper tries to set right certain ideas about the history of evolutionary developmental biology. The main point is, that we had to enface the dominance of a comparative approach towards evolutionary developmental biology before 1900, which even later on was effective in Russia, for example, till the 1930s. The problem of the experimentalist approach set against this tradition was and is that there is no concept of gestalt that may allow to integrate the former comparative views and the modern mechanistic interpretations. We argue, that it would be wrong just to describe the comparative tradition as being outdated, as it may allow to get the framework for a dynamical concept of Gestalt that may integrate the ideas of morphogenesis and pattern formation worked out in evo-devo recently. PMID- 17412295 TI - Vaccination infection and autoimmunity: myth and reality VIAMR 2005-10-26-28, Beau-Rivage Palace Hotel, Lausanne, Switzerland. PMID- 17412296 TI - Cure of chronic viral infection by neutralizing antibody treatment. AB - Persistent infections can pose severe health risks. In many cases individuals fail to clear the virus and consequently chronic infection, often associated with severe immunopathology, occurs. For some chronic infections it is known that systemic IL-10 production can be increased. However, it is unclear, whether IL-10 affects the outcome of infection, amount of immunopathology and could be the actual cause for persistence. Conventional interferons (IFN) or anti-viral immunotherapies attempting to augment anti-viral immunity directly in persistent infected individuals have failed to affect the outcome so far, but lowering the viral antigenic load has clear beneficial effects. Therefore, approaching the problem from a different angle is important. The use of neutralizing antibodies is one of the most successful methods to interfere with receptor-ligand interactions in vivo. Optimal designs of therapeutic antibody treatment regimen are important in order to achieve maximal effects with minimal duration of immunosuppression or hyperactivation. In this review we reflect on the effects of a short time administration of a neutralizing cytokine receptor antibody treatment of a chronic infection resulting in the establishment of anti-viral immune responses and viral clearance and speculate on the potential mechanisms involved in this successful therapeutic treatment. PMID- 17412297 TI - Antigenic complementarity in the induction of autoimmunity: a general theory and review. AB - The mechanism by which tolerance is broken in the induction of autoimmunity is unknown. Simple, well-characterized antigens suggest that molecular complementarity may play a key role in breaking tolerance. Experimental allergic encephalomyelitis can be induced using myelin basic protein combined with muramyl dipeptide. These molecules bind specifically to each other. Insulin antibodies can be induced when insulin is combined with glucagon, to which it binds. These cases suggest that molecular complementarity may alter the processing of "self" proteins. Antigenic complementary yields molecularly complementary immune responses (i.e., idiotypic-anti-idiotypic), undermining immune system regulation. In addition, complementarity insures that the antibodies (or T cells) directed against one antigen will molecularly mimic the other antigen, and vice versa, so that "self" and "nonself" will be confused. If at least one complementary antigen mimics a "self" protein, then an unregulated, self-sustaining immune response against tissue results. This testable theory of antigenic complementarity in autoimmunity is reviewed. PMID- 17412298 TI - Human parvovirus B19 infection and antiphospholipid antibodies. AB - Erythema infectiosum is the main manifestation of human parvovirus B19 infections. Further B19-related diseases commonly associated with the acute infection are flue-like symptoms, transient aplastic crisis, transient arthralgias, leukopenia and thrombocytopenia, spontaneous abortion and hydrops fetalis in pregnant women. Hepatitis, myocarditis, meningitis, encephalitis as well as pure red cell anemia may occur occasionally. In addition parvovirus B19 infections have been frequently described as cause or trigger of various forms of autoimmune diseases affecting all blood cell lines, joints, connective tissue, uvea, large and small vessels. Molecular mimicry may be one major contribution to the appearance of autoimmune antibodies, f.e. antiphospholipid and antineutrophil cytoplasmic antibodies as well as antinuclear antigens. These mechanisms implicated in the pathogenesis of parvovirus B19 triggered autoimmune diseases, especially focused on the development of antiphospholipid antibodies will be discussed in this short review. PMID- 17412299 TI - Psoriasis--a possible candidate for vaccination. AB - The clinical association of streptococcal infections and psoriasis is well established. The recent finding that the T cells in psoriasis skin respond to streptococcal peptidoglycan now suggests a pathway for an adaptive immune response to the streptococcal organism. These observations may allow for possible vaccines to be developed for psoriasis. PMID- 17412300 TI - Non-redundant peptidomes from DAPs: towards "the vaccine"? AB - Experimental analyses and literature survey reveal low-redundancy to the host proteins as a common denominator of immunogenic sequences mapped along tumor-, autoimmune-, and infectious disease-associated-proteins. The hypothesis that immunogenicity of peptide sequences is linked to proteomic redundancy is discussed. PMID- 17412301 TI - Human natural killer lymphocytes through the engagement of natural cytotoxicity receptors and NKG2D can trigger self-aggression. AB - NK cell killing of autologous dendritic cells (DC) is mediated by the LFA1 dependent activation of the calcium-calmodulin kinase II, resulting in degranulation and extracellular release of perforin and granzymes. On the other hand, DC killing can also be triggered by the engagement of the phosphatidylinositol 3-kinase upon ligation of the natural cytotoxicity receptors (NCR) NKp30 and NKp46. Furthermore, NK cells can also damage autologous stromal cells (SC) derived from bone marrow (BMSC) or fibroblasts obtained from skin biopsies (SF). Binding of ICAM1, expressed by BMSC or SF, to its receptor, the integrin LFA1 on NK cells, plays a key role in SC/NK interaction. Both types of SC bear the MHC-related molecule MIC-A and the UL16 binding protein ULBP3, which engagement with NKG2D, expressed by NK, is responsible for the delivery of lethal hit. HLA-I molecules do not protect DC or SC from NK cell-mediated injury. Thus, NK cells can be activated upon binding with autologous DC or SC, using different receptor-ligand pairs. On one hand this activation leads to the amplification of the NK arm of innate immune response; on the other hand, the lack of HLA-I mediated control of such activation may lead to self-reactions elicited by NK cells. PMID- 17412302 TI - Influenza vaccination in systemic lupus erythematosus: safe and protective? AB - Patients with systemic lupus erythematosus (SLE) show decreased immune responsiveness and are vulnerable for infectious diseases, due to the underlying disease and the frequent use of immunosuppressive drugs. Influenza has a high incidence in the population and is associated with increased morbidity and mortality in immunocompromised patients. Therefore, routine influenza vaccination of SLE patients seems indicated. However, there have been concerns about the safety of influenza vaccination in SLE as vaccination was thought to activate the autoimmune response. Safety of influenza vaccination has been studied, and, as far as SLE patients with quiescent disease are concerned, it is now generally accepted that influenza vaccination is safe. Another point of concern is vaccine efficacy. In immunocompromised patients, the immunogenicity of vaccines may be reduced. In the immune response to influenza (vaccination) both humoral and cell mediated responses are involved. In SLE, research on the immune response to influenza vaccination has focused on humoral immune responses, demonstrating a blunted humoral response. Future research should focus on cell-mediated immune responses as well, as these are important for clearing of influenza infection and are expected to be impaired in SLE. Because of the decreased immunogenicity of the current influenza vaccine in SLE, new influenza vaccination strategies should be explored to improve vaccination efficacy. PMID- 17412303 TI - Animal models for autoimmune hepatitis. AB - The liver is the target of adverse immune reactions in three putative autoimmune diseases: autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). These three diseases can be distinguished by clinical, histological, and immunological features. However, especially on the level of specific antibody formation an overlap can occur, which sometimes complicates diagnosis. In this review, we will concentrate on autoimmune hepatitis and the current state of animal models for this severe disease. AIH is characterized by the presence of interface hepatitis and portal plasma cell infiltration, hypergammaglobulinemia, and autoantibodies. The hallmark of type 2 AIH is the generation of autoantibodies of the LKM-1 type. The major target of these antibodies is the cytochrome P450 isoform 2D6 (CYP2D6). In the past several attempts have been made to develop a reliable animal model that reflects the persistent hepatic destruction that occurs in human AIH. However, most models were only successful in causing a transient form of hepatic damage and often used rather complex ways of disease induction. PMID- 17412304 TI - Safety and efficacy of vaccination against streptococcus pneumonia in patients with rheumatic diseases. AB - Vaccination against streptococcus pneumonia is currently recommended for patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). Safety and efficacy issues of vaccination in patients suffering from rheumatic diseases are still unresolved. This review summarizes the studies performed on the safety and immunogenicity of pneumococcal vaccination in patients with RA and SLE, with special emphasis on the effect of immunosuppressive drugs on the efficacy of the vaccine. Several trials have shown that the vaccine does not induce clinical exacerbation of RA and that it does induce an adequate humoral response, albeit one lower than that in healthy controls. PMID- 17412305 TI - Moving towards efficient therapies in type 1 diabetes: to combine or not to combine? AB - Every year, thirty thousand people worldwide are diagnosed with type 1 diabetes mellitus (T1DM). T1DM, also called autoimmune diabetes, is a multifactorial disease affecting predisposed individuals and involving genetic susceptibilities, environmental triggers, as well as unbalanced immune responses. Auto-reactive T cells, produced during the pathogenesis, play an important role by specifically destroying the pancreatic insulin-producing beta-cells in the islets of Langerhans. Numerous therapeutic interventions have been tested, mostly in animal models, but also in humans. To date, only three phase II/III clinical trials have demonstrated safety and efficacy: anti-CD3 antibody, DiaPep277, and GAD65 (in patients with latent autoimmune diabetes in adults). Unfortunately, a significant number of patients did not respond positively and remained insulin-dependent after completion of therapy. Several reasons account for this. Firstly, the severity of the disease as well as the auto-aggressive T cell repertoire vary from patient to patient leading to a broad range of therapeutic efficacies, and secondly at the time of the treatment the number of remaining beta-cells will directly impact the level of insulin production post-treatment. In this review, we will provide some clues to enhance efficacy of future immuno-interventions in patients with T1DM. We suggest that combination therapies might be the best approach. PMID- 17412306 TI - Estrogen esters as substrates for human paraoxonases. AB - Mammalian paraoxonases (PONs 1, 2 and 3) are a highly conserved family of esterases, with uncertain physiological functions and natural substrates. Here we characterize the ability of purified recombinant human PONs to hydrolyze estrogen esters, a class of compounds previously not known to be PON substrates. PONs hydrolyzed estrogen mono- and diesters at position 3 of the steroid A-ring. Diesters were better substrates for the PONs and were very efficiently hydrolyzed, particularly by PON3. Esters at position 17 were not cleaved by the PONs unless an adjacent double bound was present. Purified human serum butyryl cholinesterase also hydrolyzed estrogen esters, however it preferably hydrolyzed the mono-esters. The ability of the PONs' to effectively hydrolyze a variety of estrogen esters provides further insight into the structure of their active sites and suggests that natural compounds with aromatic ester groups might be relevant substrates for the PONs. PMID- 17412307 TI - Nanomechanical and chemical characterization of incipient in vitro carious lesions in human dental enamel. AB - OBJECTIVE: The research was designed to examine the growth of in vitro carious lesions in dental enamel using nanoindentation and time-of-flight secondary ion mass spectroscopy (TOF-SIMS). This was intended to give maps of mechanical properties and chemistry over the cross-section of the lesions. METHODS: Incipient carious lesions were grown on the buccal faces of 20 human premolars by exposure to acid for 3, 5, 7 or 14 days. The lesions were then cut in cross section normal to the exposed surface. The lesions' cross-sections were then examined using nanoindentation and TOF-SIMS. RESULTS: The earliest lesions (3 days of acid exposure) showed little evidence of lesion growth, but the 5, 7 and 14 days of exposure all gave lesions with a weak, demineralized interior, but a stronger, less demineralized surface zone. The thickness of the surface zone was found to diminish with the length of exposure to acid, but it was still present even after 14 days of exposure. CONCLUSION: The results indicate that carious lesions develop subsurface and that the surface zone forms by a coupled diffusion process. Mechanically the lesion has a strong surface layer, but a very weak interior which makes the lesion vulnerable to mechanical loading. However, the presence of a surface zone that retains a high mineral content and is mechanically strong suggests that lesion development can be arrested and possibly reversed even when the lesions are relatively mature. PMID- 17412308 TI - Photosynthetic electron transport activity in heat-treated barley leaves: the role of internal alternative electron donors to photosystem II. AB - Electron transport processes were investigated in barley leaves in which the oxygen-evolution was fully inhibited by a heat pulse (48 degrees C, 40 s). Under these circumstances, the K peak (approximately F(400 micros)) appears in the chl a fluorescence (OJIP) transient reflecting partial Q(A) reduction, which is due to a stable charge separation resulting from the donation of one electron by tyrozine Z. Following the K peak additional fluorescence increase (indicating Q(A)(-) accumulation) occurs in the 0.2-2 s time range. Using simultaneous chl a fluorescence and 820 nm transmission measurements it is demonstrated that this Q(A)(-) accumulation is due to naturally occurring alternative electron sources that donate electrons to the donor side of photosystem II. Chl a fluorescence data obtained with 5-ms light pulses (double flashes spaced 2.3-500 ms apart, and trains of several hundred flashes spaced by 100 or 200 ms) show that the electron donation occurs from a large pool with t(1/2) approximately 30 ms. This alternative electron donor is most probably ascorbate. PMID- 17412309 TI - Th1 and Th2 cells are required for both eosinophil- and neutrophil-associated airway inflammatory responses in mice. AB - Most current animal models focus on eosinophil-mediated asthma, despite compelling evidence that a neutrophil-mediated disease occurs in some asthma patients. Using intranasal challenge of mice sensitized either orally or nasally with whole peanut protein extract in the presence of cholera toxin, we developed mouse models of eosinophil- and neutrophil-mediated asthma, respectively. In this study, mice deficient in Th1 (IL-12 and IFN-gamma) or Th2 (IL-4 and IL-13) pathways were used to characterize the role played by Th1 and Th2 cytokines during the initial priming phase in the two models. Antigen-specific Ab responses were controlled primarily by Th2 cytokines in mice sensitized by the oral route, whereas Th1 cytokines appeared to play a predominant role in mice sensitized by the nasal route. Furthermore, the absence of key Th1 or Th2 cytokines during the initial phase of priming reduced lung reactivity in both mouse models of airway inflammation. PMID- 17412310 TI - Neural mechanisms underlying immediate and final action goals in object use reflected by slow wave brain potentials. AB - Event-related brain potentials were used to study the neural mechanisms underlying goal-directed object use distinguishing between processes supporting immediate and final action goals during action planning and execution. Subjects performed a grasping and transportation task in which actions were cued either with the immediate action goal (the part of the object to grasp) or with the final action goal of the movement (the end position for transportation). Slow wave potentials dissociated between processes supporting immediate and final goals: reaching for the object was accompanied by the development of a parietal occipital slow wave that peaked in congruency with the grasping event, whereas transport of the object towards the final goal location was found accompanied by slow wave components developing over left frontal regions with a peak towards the movement end. Source localization of cueing differences indicated activation centered around the parieto-occipital sulcus during reaching of the immediate action goal, followed by enhanced activation in the anterior prefrontal cortex during transport to the final action goal. These results suggest the existence of separate neural controllers for immediate and final action goals during the execution of goal-directed actions with objects. PMID- 17412311 TI - Study on glycosylated prodrugs of toxoflavins for antibody-directed enzyme tumor therapy. AB - Eight novel toxoflavin glycosides, which are potential prodrugs in antibody directed enzyme prodrug therapy (ADEPT), were synthesized. The structures of all toxoflavin glycosides were characterized by (13)C NMR spectroscopy, elemental analysis, and MS. Their enzymatic hydrolysis activities were tested against beta glucosidase (EC.3.2.1.21). PMID- 17412312 TI - Local adipose tissue depots as cardiovascular risk factors. AB - Obesity is associated with increased cardiovascular morbidity and mortality. Although obesity-associated hypertension, dyslipidemia and insulin resistance account in part for this association, it becomes increasingly apparent that a systemic and local pro-inflammatory response of adipose tissue might also be a contributing factor. White adipose tissue (WAT) is a highly active organ secreting various peptides such as cytokines, chemokines and hormone-like proteins. Besides the visceral and subcutaneous depots, WAT is also found in the close vicinity of blood vessels (perivascular adipose tissue), where it secretes cytokines such as interleukin-1, tumor necrosis factor alpha, pro-atherogenic chemokines, and pro-angiogenic peptides. These factors appear to contribute directly to alterations of the function and structure of the vascular wall, including chronic inflammation, alterations of vascular tone, proliferation of smooth muscle cells, neo-angiogenesis and hence to the development of atherosclerosis and cardiovascular complications. PMID- 17412313 TI - The microenvironment can shift erythrocytes from a friendly to a harmful behavior: pathogenetic implications for vascular diseases. AB - Erythrocytes are peculiar cells aimed at the delivery of oxygen and nitric oxide to the periphery and carbon dioxide to the lungs. In addition, they also exert, under physiological conditions, a scavenging activity towards reactive oxygen and nitrogen species often over-produced in morbidity states, e.g. in inflamed tissues. Their deformability is essential for their circulation, specifically in small blood vessels, and this is an important pre-requisite for such vascular "antioxidant" functions. On the other hand, if the erythrocyte undergoes changes in its redox status, i.e. is not capable of counteracting the pro-oxidant status of the microenvironment, it becomes a source of reactive species and, consequently, its typical structural and functional features are lost. More importantly, the oxidatively modified red cell increases its aggregability and adhesiveness to the endothelium and to other blood cells, thus contributing to vascular damage. In line with recent data from the literature, erythrocytes can be proposed as bioindicators of progression in chronic or acute diseases characterized, as a hallmark, by oxidative alterations. PMID- 17412314 TI - The acute phase protein alpha2-macroglobulin induces rat ventricular cardiomyocyte hypertrophy via ERK1,2 and PI3-kinase/Akt pathways. AB - OBJECTIVE: Alpha2-macroglobulin (alpha2M) is an acute phase protein released to the serum upon challenges such as cardiac hypertrophy and infarction. Here we report on the role of alpha2M in the induction of hypertrophic cell growth, contractile responsiveness of rat ventricular cardiomyocytes, and on the underlying extracellular regulated kinase 1,2 (ERK1,2) and phosphoinositide 3 kinase (PI3-kinase)/Akt pathways. METHODS: Cell volume and cross-sectional areas were assessed as parameters of hypertrophic growth, and real time RT-PCR for the analysis of hypertrophy-related genes was performed. Protein synthesis was analyzed by 14C-phenylalanine incorporation. Activation of ERK1,2, PI3-kinase and Akt was assessed by immunohistochemical analysis of phosphorylated proteins. Contractile responsiveness was investigated by determination of cell shortening following electrical field stimulation. Intracellular calcium concentration [Ca2+]i was determined by fluo-3 microfluorometry. RESULTS: Treatment of ventricular cardiomyocytes for 24 h with alpha2M significantly increased cell volume and protein synthesis as well as expression of hypertrophy-associated genes [brain natriuretic protein (BNP), beta-myosin heavy chain (beta-MHC), myosin light chain-2 (MLC-2), atrial natriuretic factor (ANF), and skeletal alpha actin]. Comparable effects were achieved by treatment of cells with an antibody directed against the alpha2M-receptor LDL receptor-related protein-1 (LRP-1) and counteracted upon coincubation with receptor-associated protein (RAP), suggesting an involvement of alpha2M-LRP-1 signalling. Furthermore, alpha2M treatment increased sarcoplasmic reticulum Ca2+-ATPase (SERCA-2a) expression, diastolic and systolic [Ca2+]i, and contractile responsiveness after electrical stimulation. Shortly after alpha2M stimulation, activation of ERK1,2, Akt, and PI3-kinase pathways was observed. Consequently, alpha2M-induced protein synthesis was inhibited upon treatment with the ERK1,2 inhibitor UO126 as well as by LY294002 and wortmannin, which inhibit PI3-kinase, and by rapamycin, which inhibits mammalian target of rapamycin (mTOR) downstream of Akt. CONCLUSIONS: Our data show that alpha2M induces hypertrophic cell growth in rat ventricular cardiomyocytes via ERK1,2 and PI3-kinase/Akt and improves cardiac cell function. PMID- 17412315 TI - Nitric oxide and superoxide: interference with hypoxic signaling. AB - Sensing and responding to changes in oxygen partial pressure assures that the cellular oxygen supply is tightly controlled in order to balance the risks of oxidative damage vs. oxygen deficiency. The hypoxia inducible factor (HIF) regulatory system is controlled by prolyl hydroxylases (PHDs), the von Hippel Lindau protein (pVHL), and the 26S proteasome and transduces changes in oxygenation to adequate intracellular adaptive responses. A functional HIF response requires stabilization of the alpha-subunit, e.g. HIF-1alpha, during hypoxia and dimerization with HIF-1beta, to drive target gene activation. Intriguingly, high concentrations of nitric oxide (NO) stabilize HIF-1alpha and thus mimic a hypoxic response under normoxia. Mechanistically, NO blocks PHD activity and attenuates proline hydroxylation of HIF-1alpha. This causes dissociation of pVHL from HIF-1alpha and, consequently, HIF-1alpha accumulates because proteasomal destruction is impaired. However, during hypoxia low concentrations of NO facilitate destruction of HIF-1alpha and thus reverse HIF signaling. Under these conditions, NO impairs respiration and avoids oxygen gradients that limit PHD activity. An additional layer of complexity comprises the interaction of NO with O(2)(-). Signaling qualities attributed to NO are antagonized by compensatory flux rates of O(2)(-) and vice versa to adjust levels of HIF-1alpha under normoxia and hypoxia. The liaison of NO and hypoxia is versatile and ranges from courting to matrimony and divorce. PMID- 17412316 TI - Long-term fenofibrate treatment impairs endothelium-dependent dilation to acetylcholine by altering the cyclooxygenase pathway. AB - OBJECTIVE: Experimental studies and opinion articles emphasize that cardiovascular alterations associated with ageing can be improved by the long term use of fenofibrates. We analyzed the effect of fenofibrate treatment on the acetylcholine-induced relaxation in rat aorta and the participation of nitric oxide (NO) and cyclooxygenase (COX)-derived factors in this effect. METHODS: Acetylcholine relaxation in untreated and 6-week fenofibrate-treated Wistar rats was analyzed in the absence and presence of the NO synthase (NOS) inhibitor N(G) nitro-L-arginine methyl ester (L-NAME), the specific inducible NO (iNOS) synthase inhibitor 1400W, the nonspecific COX inhibitor indomethacin, the specific COX-2 inhibitor NS-398, the specific thromboxane receptor antagonist SQ-29548, the thromboxane synthesis inhibitor furegrelate, the prostacyclin synthesis inhibitor tranylcypromine, or the 20-HETES synthesis inhibitor formamidine. eNOS, iNOS, COX 1, and COX-2 expression was studied by Western blotting. In addition, production of prostaglandin F(2alpha) (PGF(2alpha)), thromboxane A(2) (TxA(2)), prostaglandin E(2) (PGE(2)), isoprostanes, and prostacyclin (PGI(2)) was also measured. RESULTS: Fenofibrate treatment reduced acetylcholine relaxation. Indomethacin, NS-398, and tranylcypromine decreased acetylcholine relaxation in untreated rats but enhanced relaxation in treated rats. SQ-29548 increased acetylcholine responses in segments from treated rats but not in segments from untreated rats. L-NAME decreased vasodilator response to acetylcholine in both groups while furegrelate, NS-398, 1400W, and formamidine did not affect acetylcholine responses in either group. eNOS and COX-2 expression was higher in aorta from treated rats while COX-1 and iNOS remained unmodified. Basal and acetylcholine-stimulated NO and PGE(2) release were increased, and that of PGI(2) decreased in treated rats. TxA(2) release was similar, but PGF(2alpha) release was undetectable in both groups. CONCLUSIONS: Although it increases NO production through increases in eNOS expression, fenofibrate treatment induces endothelial dysfunction. This effect seems to be mediated by decreased PGI(2) and increased PGE(2) release, and it may help to explain the rise in thromboembolic events observed after long-term fenofibrate treatment in humans. PMID- 17412317 TI - Interaction of dipalmitoyl phosphatidylserine with ethanol: induction of an ordered gel phase at room temperature. AB - Using differential scanning calorimetry and small and wide-angle X-ray diffraction, we show that, unlike the saturated phosphatidylcholines, for which ethanol induces chain interdigitation in the gel state, and unlike natural phosphatidylserine in which the gel state is almost unaffected by the addition of ethanol, dipalmitoyl phosphatidylserine (DPPS) assumes an ordered structure after incubation at room temperature in the presence of as little as 5% (v/v) ethanol. In the liquid crystalline state, a progressive decrease in the interbilayer spacing is observed as a function of ethanol concentration, similar to what is found for natural phosphatidylserine (PS) and 1-palmitoyl-2-oleoyl phosphatidylserine (POPS). The 0.37 molar fraction of cholesterol in the DPPS dispersion in the presence of 10% (v/v) ethanol, does not prevent the formation of the ordered gel. PMID- 17412318 TI - Paracrine and autocrine mechanisms of apelin signaling govern embryonic and tumor angiogenesis. AB - Apelin and its G protein-coupled receptor APJ play important roles in blood pressure regulation, body fluid homeostasis, and possibly the modulation of immune responses. Here, we report that apelin-APJ signaling is essential for embryonic angiogenesis and upregulated during tumor angiogenesis. A detailed expression analysis demonstrates that both paracrine and autocrine mechanisms mark areas of embryonic and tumor angiogenesis. Knockdown studies in Xenopus reveal that apelin-APJ signaling is required for intersomitic vessel angiogenesis. Moreover, ectopic expression of apelin but not vascular endothelial growth factor A (VEGFA) is sufficient to trigger premature angiogenesis. In vitro, apelin is non-mitogenic for primary human endothelial cells but promotes chemotaxis. Epistasis studies in Xenopus embryos suggest that apelin-APJ signaling functions downstream of VEGFA. Finally, we show that apelin and APJ expression is highly upregulated in microvascular proliferations of brain tumors such as malignant gliomas. Thus, our results define apelin and APJ as genes of potential diagnostic value and promising targets for the development of a new generation of anti-tumor angiogenic drugs. PMID- 17412319 TI - The Caenorhabditis elegans pumilio homolog, puf-9, is required for the 3'UTR mediated repression of the let-7 microRNA target gene, hbl-1. AB - The Puf family of RNA-binding proteins directs cell fates by regulating gene expression at the level of translation and RNA stability. Here, we report that the Caenorhabditis elegans pumilio homolog, puf-9, controls the differentiation of epidermal stem cells at the larval-to-adult transition. Genetic analysis reveals that loss-of-function mutations in puf-9 enhance the lethality and heterochronic phenotypes caused by mutations in the let-7 microRNA (miRNA), while suppressing the heterochronic phenotypes of lin-41, a let-7 target and homolog of Drosophila Brat. puf-9 interacts with another known temporal regulator hbl-1, the Caenorhabditis elegans ortholog of hunchback. We present evidence demonstrating that puf-9 is required for the 3'UTR-mediated regulation of hbl-1, in both the hypodermis and the ventral nerve cord. Finally, we show that this regulation is dependent on a region of the hbl-1 3'UTR that contains putative Puf family binding sites as well as binding sites for the let-7 miRNA family, suggesting that puf-9 and let-7 may mediate hypodermal seam cell differentiation by regulating common targets. PMID- 17412320 TI - Bifurcation dynamics in lineage-commitment in bipotent progenitor cells. AB - Lineage specification of multipotent progenitor cells is governed by a balance of lineage-affiliated transcription factors, such as GATA1 and PU.1, which regulate the choice between erythroid and myelomonocytic fates. But how ratios of lineage determining transcription factors stabilize progenitor cells and resolve their indeterminacy to commit them to discrete, mutually exclusive fates remains unexplained. We used a simple model and experimental measurements to analyze the dynamics of a binary fate decision governed by a gene-circuit containing auto stimulation and cross-inhibition, as embodied by the GATA1-PU.1 paradigm. This circuit generates stable attractors corresponding to erythroid and myelomonocytic fates, as well as an uncommitted metastable state characterized by coexpression of both regulators, explaining the phenomenon of "multilineage priming". GATA1 and PU.1 mRNA and transcriptome dynamics of differentiating progenitor cells confirm that commitment occurs in two stages, as suggested by the model: first, the progenitor state is destabilized in an almost symmetrical bifurcation event, resulting in a poised state at the boundary between the two lineage-specific attractors; second, the cell is driven to the respective, now accessible attractors. This minimal model captures fundamental features of binary cell fate decisions, uniting the concepts of stochastic (selective) and deterministic (instructive) regulation, and hence, may apply to a wider range of binary fate decision points. PMID- 17412321 TI - MTHFR Gene polymorphisms, B-vitamins and hyperhomocystinemia in young and middle aged acute myocardial infarction patients. AB - We have examined the prevalence of the C677T and A1298C single nucleotide polymorphisms (SNPs) in the methylenetetrahydrofolate reductase (MTHFR) gene in healthy Tamilians and in patients with acute myocardial infarction and related this polymorphism to plasma homocysteine concentrations, serum folate, serum cobalamin and riboflavin status. The SNPs in the MTHFR gene were determined by polymerase chain reaction-restriction fragment length polymorphism analysis. Plasma homocysteine, serum folate and serum cobalamin concentrations were analyzed using an automated chemiluminescence method and riboflavin status was assessed by measuring the erythrocyte glutathione reductase activity using spectrophotometric method. Out of the 200 young and middle-aged (<48 years) individuals included in the study, 100 were acute myocardial infarction (AMI) patients and 100 were healthy individuals with no documented history of heart diseases. There was a significant increase in homocysteine levels among the AMI patients as compared to the healthy controls (p<0.001). The results of this study indicate that hyperhomocystinemia is more prevalent in Tamilian AMI patients and that the MTHFR C677T and A1298C SNPs are not associated with hyperhomocystinemia. Folate status was found to be within normal range in all the study subjects. There was no correlation between homocysteine and different biochemical variables including cobalamin, folate and riboflavin. However, serum cobalamin was found to be significantly decreased in AMI patients when compared to controls (p<0.001). The simultaneous presence of decreased serum cobalamin status, hyperhomocystinemia and mutant genotype for both the SNPs might lead to an increased risk for the occurrence of AMI. Further intervention trials including the supplementation of cobalamin may prove whether homocysteine level decrease in response to the supplementation of cobalamin in individuals with hyperhomocystinemia and mutant genotype for both the above mentioned SNPs. PMID- 17412322 TI - Focus on molecules: lacritin. PMID- 17412323 TI - Cripto-1 expression in uveal melanoma: an immunohistochemical study. AB - Human Cripto, the founder member of the epidermal growth factor-Cripto-FRL1 Cryptic (EGF-CFC) family, plays an important role during early embryonic development and in particular in carcinogenesis and the development of cancer metastases. Cripto-1 is over-expressed in most cancers, but is absent or only weakly expressed in normal cells. For this reason, Cripto-1 could be of potential value in the targeted treatment. There is no information on the expression of Cripto-1 in human uveal melanoma. Cripto-1 reactivity was evaluated by immunohistochemistry on 36 archival uveal melanomas using the polyclonal antibody to Cripto-1. The tumors were divided in to 2 groups. There were 18 uveal melanomas with no intrascleral or extrascleral extension and 18 uveal melanomas with intrascleral/extrascleral extension/liver metastasis. Cripto-1 reactivity was correlated with tumor aggressiveness and cell type. Furthermore, we studied the immunolocalization of Cripto-1 in 4 uveal melanoma cell lines OCM-1, OCM-8, and 92-1, and OMM-1 and in 2 primary uveal melanocyte cultures. Cripto-1 was expressed in both the non-invasive and aggressive uveal melanomas. Cripto-1 was positive in the 4 uveal melanoma cell lines and absent in the primary uveal melanocyte cultures. Retinal tissue did not express Cripto-1. The results suggest that Cripto-1 is expressed in uveal melanoma, negative in the non-neoplastic ocular tissue and point to its use as a target for therapy. PMID- 17412324 TI - Plant proton pumps. AB - Chemiosmotic circuits of plant cells are driven by proton (H(+)) gradients that mediate secondary active transport of compounds across plasma and endosomal membranes. Furthermore, regulation of endosomal acidification is critical for endocytic and secretory pathways. For plants to react to their constantly changing environments and at the same time maintain optimal metabolic conditions, the expression, activity and interplay of the pumps generating these H(+) gradients have to be tightly regulated. In this review, we will highlight results on the regulation, localization and physiological roles of these H(+)- pumps, namely the plasma membrane H(+)-ATPase, the vacuolar H(+)-ATPase and the vacuolar H(+)-PPase. PMID- 17412325 TI - The aryl hydrocarbon receptor, more than a xenobiotic-interacting protein. AB - The aryl hydrocarbon (dioxin) receptor (AhR) has been studied for several decades largely because of its critical role in xenobiotic-induced toxicity and carcinogenesis. Albeit this is a major issue in basic and clinical research, an increasing number of investigators are turning their efforts to try to understand the physiology of the AhR under normal cellular conditions. This is an exciting area that covers cell proliferation and differentiation, endogenous mechanisms of activation, gene regulation, tumor development and cell motility and migration, among others. In this review, we will attempt to summarize the studies supporting the implication of the AhR in those endogenous cellular processes. PMID- 17412326 TI - Inhibition of hepatic glucose 6-phosphatase system by the green tea flavanol epigallocatechin gallate. AB - Effect of 5-100 microM epigallocatechin gallate (EGCG) on hepatic glucose 6 phosphatase (G6Pase) system was investigated. EGCG inhibited G6Pase in intact but not in permeabilized rat liver microsomes, suggesting the interference with the transport. However, EGCG did not hinder microsomal glucose 6-phosphate (G6P) uptake. Instead, it increased the accumulation of radioactivity after the addition of [(14)C]G6P, presumably due to a slower release of [(14)C]glucose, the product of luminal hydrolysis. Indeed, EGCG was found to inhibit microsomal glucose efflux. Since G6Pase activity is depressed by glucose in a concentration dependent manner, we concluded that EGCG inhibits G6Pase through an elevated luminal glucose level. PMID- 17412327 TI - Knockdown of ACAT-1 reduces amyloidogenic processing of APP. AB - Previous studies have shown that acyl-coenzyme A:cholesterol acyl transferase (ACAT), an enzyme that controls cellular equilibrium between free cholesterol and cholesteryl esters, modulates proteolytic processing of APP in cell-based and animal models of Alzheimer's disease. Here we report that ACAT-1 RNAi reduced cellular ACAT-1 protein by approximately 50% and cholesteryl ester levels by 22% while causing a slight increase in the free cholesterol content of ER membranes. This correlated with reduced proteolytic processing of APP and 40% decrease in Abeta secretion. These data show that even a modest decrease in ACAT activity can have robust suppressive effects on Abeta generation. PMID- 17412328 TI - Growth-associated protein 43-positive sensory nerve fibers accompanied by immature vessels are located in or near peritoneal endometriotic lesions. AB - OBJECTIVE: To investigate the topographical relationship between nerve fibers and peritoneal endometriotic lesions and to determine the origin of endometriosis associated nerve fibers. DESIGN: Retrospective nonrandomized study. SETTING: University hospital endometriosis research center. PATIENT(S): Premenopausal women with histologically confirmed endometriosis were selected (n = 73). Peritoneal endometriotic lesions (n = 106) and unaffected peritoneal biopsies from patients without endometriosis (n = 9) were obtained. INTERVENTION(S): Immunohistochemistry was used to study the expression of neurofilament, substance P, smooth muscle actin, von Willebrand factor, growth-associated protein 43, nerve growth factor, and neutrophin-3 in peritoneal endometriotic lesion samples from women with symptomatic endometriosis and in peritoneal samples from women without endometriosis. RESULT(S): Pain-conducting substance-P-positive nerve fibers were found to be directly colocalized with human peritoneal endometriotic lesions in 74.5% of all cases. The endometriosis-associated nerve fibers are accompanied by immature blood vessels within the stroma. Nerve growth factor and neutrophin-3 are expressed by endometriotic cells. Growth-associated protein 43, a marker of neural outgrowth and regeneration, is expressed in endometriosis associated nerve fibers but not in existing peritoneal nerves. CONCLUSION(S): The data provide the first evidence of direct contact between sensory nerve fibers and peritoneal endometriotic lesions. This implies that the fibers play an important role in the etiology of endometriosis-associated pelvic pain. Moreover, emerging evidence suggests that peritoneal endometriotic cells exhibit neurotrophic properties. PMID- 17412330 TI - Prognostic factors in oocyte donation: an analysis through egg-sharing recipient pairs showing a discordant outcome. AB - OBJECTIVE: To analyze prognostic factors that are associated with a discordant outcome in egg recipients sharing oocytes from the same donor. DESIGN: Matched case-control single-center study. SETTING: Private infertility clinic. PATIENT(S): Four hundred forty-four recipients (222 pairs) sharing oocytes from the same donor and showing a discordant outcome. INTERVENTION(S): Controlled ovarian hyperstimulation of egg donors, oocyte donation, intracytoplasmic sperm injection, and ET in egg recipients. MAIN OUTCOME MEASURE(S): Recipient age, obstetric (gravidity, parity) and gynecologic variables (previous uterine surgery, uterine fibroids, uterine malformations, endometriosis, history of tubal infertility), previous oocyte donation cycles, duration of E(2) replacement, received cumulus-oocyte complexes, mature (MII) oocytes, fertilized oocytes, transferred embryos, mean embryo score, transfer difficulty, and semen parameters. RESULT(S): No significant differences were found in the above mentioned prognostic factors between the study and control groups. CONCLUSION(S): Recipient- and cycle-related prognostic factors investigated in our study were not associated with a discordant outcome in recipient pairs sharing oocytes from the same donor. Other possible prognostic factors involving oocyte donor heterogeneity, embryo aneuploidy rates, male factor infertility, and endometrial receptivity should be further investigated. PMID- 17412329 TI - Effect of a daily supplement of soy protein on body composition and insulin secretion in postmenopausal women. AB - OBJECTIVE: To determine whether a supplement of soy protein improves body composition, body fat distribution, and glucose and insulin metabolism in postmenopausal women without diabetes compared with an isocaloric casein placebo. DESIGN: Randomized, double-blind, placebo-controlled 3-month trial. SETTING: Clinical Research Center. PATIENT(S): Fifteen postmenopausal women. INTERVENTION(S): Computed tomographic scans at L4/L5, dual energy x-ray absorptiometry, hyperglycemic clamps. MAIN OUTCOME MEASURE(S): Total fat, total abdominal fat, visceral fat, subcutaneous abdominal fat, and insulin secretion. RESULT(S): Weight by dual energy x-ray absorptiometry did not change between groups (+1.38 +/- 2.02 kg for placebo vs. +0.756 +/- 1.32 kg for soy, mean +/- SD). Total and subcutaneous abdominal fat increased more in the placebo group than in the soy group (for differences between groups in total abdominal fat: +38.62 +/- 22.84 cm(2) for placebo vs. -11.86 +/- 31.48 cm(2) for soy; subcutaneous abdominal fat: +22.91 +/- 28.58 cm(2) for placebo vs. -14.73 +/- 22.26 cm(2) for soy). Insulin secretion, visceral fat, total body fat, and lean mass did not differ between groups. Isoflavone levels increased more in the soy group. CONCLUSION(S): A daily supplement of soy protein prevents the increase in subcutaneous and total abdominal fat observed with an isocaloric casein placebo in postmenopausal women. PMID- 17412331 TI - Use of pig oocytes for training new professionals in human assisted reproduction laboratories. AB - OBJECTIVE: To evaluate whether swine oocytes are useful for training new technicians in a human reproduction laboratory. DESIGN: Prospective study. SETTING: Graduate school in assisted reproductive techniques (ART) in Brazil. PATIENT(S): Students in a human reproduction laboratory. INTERVENTION(S): Medium sized follicles were aspirated from prepubertal gilts' ovaries and collected at a slaughterhouse. Oocytes were retrieved from the follicular fluid. Twenty-one students trained during four periods of 20 hours each were evaluated as to their ability to perform micromanipulation and were compared with a group of well trained professionals (control group). MAIN OUTCOME MEASURE(S): Students' ability in oocyte retrieval, oocyte manipulation, and intracytoplasmic sperm injection during and after the 80 hours of training. RESULT(S): Students were able to retrieve, on average, 23.8 oocytes per ovary. Their micromanipulation skills substantially increased, reaching an oocyte retrieval rate of 77.2%, compared with 84.9% in the control group after the training period. Although the oocyte damage rate gradually decreased, from 52.0% after 20 hours of training to 5.4% after 80 hours, these figures were still above the control group oocyte damage rate by 0.3%, which was a statistically significant level. Regarding intracytoplasmic sperm injection, within 40 hours, no students were able to perform a single injection; and by the end of 80 hours, they achieved an average of 4.0 oocytes per hour, whereas the control group injected 20.6 oocytes per hour, a statistically significant difference. CONCLUSION(S): Swine ovaries may be a useful tool in the spectrum of training techniques for unskilled assisted reproductive techniques laboratory professionals. PMID- 17412332 TI - Simple tools for assessment of ovarian reserve (OR): individual ovarian dimensions are reliable predictors of OR. AB - OBJECTIVE: To determine the association between individual ovarian dimensions, advancing age, and declining ovarian reserve (OR) in an infertile population. DESIGN: An ongoing prospective observational study. SETTING: Academic infertility practice. PATIENT(S): Sixty-nine premenopausal women presenting for the evaluation and management of infertility. INTERVENTION(S): Transvaginal ultrasound assessment of the ovarian dimensions (length, width, and overall diameter in cm) and evaluation of the OR status by measuring serum levels of FSH and E(2) in the early follicular phase of the menstrual cycle. MAIN OUTCOME MEASURE(S): Relationship of the individual ovarian dimensions to age (years) and OR (reflected by historical maximal FSH levels). RESULT(S): A statistically significant decrease in three ovarian size parameters accompanied advancing age (ovarian width, r = -0.30; ovarian length, r = -0.24; and the mean overall ovarian diameter, r = -0.30). Levels of FSH demonstrated a significant and linear correlation with age (r = 0.39). Increasing levels of FSH (and hence declining OR) were associated with statistically significant declines in the mean ovarian width (r = -0.39), length (r = -0.38), and overall mean ovarian diameter (r = .42), Patients with a known diagnosis of diminished OR demonstrated significantly reduced ovarian dimensions compared with patients with other infertility etiologies. Multivariate linear regression analysis confirmed individual ovarian measurements (width, length, and overall diameter) as independent predictors of OR (FSH levels) after adjusting for parameters that are known to influence ovarian size, i.e., age, smoking status, body mass index, and anovulation history. Ovarian width emerged as the strongest predictor of OR (largest beta coefficient and hence the strongest association with OR status), compared with ovarian length and the mean ovarian diameter. CONCLUSION(S): Single ovarian dimensions are reliable predictors of advancing age and declining OR status in premenopausal infertile women. The magnitude of this association is most robust for the ovarian width. PMID- 17412333 TI - Treatment of heterotopic cervical pregnancy after in vitro fertilization-embryo transfer by using transvaginal ultrasound-guided aspiration and instillation of hypertonic solution of sodium chloride. AB - OBJECTIVE: To report a case of successful treatment of heterotopic cervical pregnancy resulting from IVF-embryo transfer. DESIGN: Case report. SETTING: Tertiary university clinical center. PATIENT(S): A 31-year-old patient, treated for heterotopic cervical pregnancy, diagnosed in the 6th gestational week. INTERVENTION(S): Transvaginal ultrasound-guided aspiration of cervical pregnancy and instillation of hypertonic solution of sodium chloride, upon ligation of descending cervical branches of the uterine arteries. MAIN OUTCOME MEASURE(S): Patient recovery and further maintenance of intrauterine twin pregnancy. RESULT(S): The cervical pregnancy was successfully aborted, and the intrauterine twin pregnancy was successfully maintained. CONCLUSION(S): The intervention applied may be used in treatment of heterotopic cervical pregnancy. PMID- 17412334 TI - Hysteroscopic management of lower genital tract lesions in females with limited vaginal access. AB - OBJECTIVE: To show that hysteroscopy is a feasible and safe method of evaluation in females with limited vaginal access. DESIGN: Retrospective review of two patients who received hysteroscopy for evaluation of uterus didelphys with hemivagina and a cervical polyp. SETTING: Private hospital. PATIENT(S): Two patients with either a vagina septum or cervical polyp. INTERVENTION(S): Hysteroscopy for visualization and resection. MAIN OUTCOME MEASURE(S): Resection of a vagina septum and cervical polyp. RESULT(S): No more cyclic dysmenorrhea in the patient with uterus didelphys and cessation of abnormal bleeding in the patient with a cervical polyp. Our results suggest that hysteroscopy can provide precise visualization of vagina and cervical lesions if the approach is difficult with decreased access, and allows concurrent treatment of pathologic conditions accordingly. CONCLUSION(S): Hysteroscopy is favorable in the diagnosis and treatment of vaginal and cervical lesions in women with limited vaginal access because of age or a narrower vagina. In the Eastern culture, it is very important for females to maintain their state of virginity signified by an uninjured hymen ring. This problem is solved, as hysteroscopy allows the evaluation of malformations of the female genital tract. PMID- 17412335 TI - How viable are zygotes in which the PN are still intact at 25 hours? Impact on the choice of embryo for transfer. AB - OBJECTIVE: To evaluate the pertinence of kinetic observations of embryonic development 25 hours after IVF, with a view to optimizing the choice of embryos for transfer. DESIGN: Retrospective study of 471 cycles with embryo transfer. SETTING: University Hospital. INTERVENTION(S): Observations of changes in zygotes, in the 25th hour after insemination (25 h PI). MAIN OUTCOME MEASURE(S): Identification of three stages of development: two cells (2C), loss of the pronuclei (in syngamy; 0PN) and persistence of the two pronuclei (2PN). Embryonic morphology at 48 hours, according to the stages identified at 25 hours, was correlated with the rates of implantation and clinical pregnancy. RESULT(S): Two thousand four hundred seventy-eight day-2 embryos were observed. The number of blastomeres on day 2 differed significantly according to stage of development at 25 hours (2C: 69.8% vs. 0PN: 55% and 2PN: 24.6%). Implantation rates were significantly lower for cycles in which only 2PN embryos were produced (7%, n = 160) than for cycles with at least one 2C embryo and/or, preferably, a 0PN embryo (14.4%). The implantation rate was significantly lower for the transfer of only 2PN embryos (7.6%) than for the transfer of only embryos that had advanced further by 25 h PI (15.2%). If only one type of embryo was transferred (2PN vs. 2C or 0PN), for embryos with a similar morphology at 48 hours, four-blastomere embryos derived from 2PN embryos implanted less efficiently than those derived from 2C and/or 0PN embryos. CONCLUSION(S): Observation at 25 h PI is a simple, rapid, and cheap method for identifying the best embryos to transfer. A lack of zygote development at 25 hours is associated with a poor prognosis, unless the zygote concerned has two to three cells on day 2. The development of zygotes with 2PN at 25 h PI should be taken into account as a function of their development at 48 hours. PMID- 17412336 TI - The beneficial effects of toremifene administration on the hypothalamic-pituitary testicular axis and sperm parameters in men with idiopathic oligozoospermia. AB - OBJECTIVE: To evaluate whether toremifene, a selective estrogen receptor modulator (SERM), has a beneficiary effect on all three main sperm parameters. DESIGN: Prospective interventional clinical study. SETTING: University hospital. PATIENT(S): One-hundred subfertile men with idiopathic oligozospermia. INTERVENTION(S): Toremifene (60 mg daily) was administered to all men for 3 months. At baseline and at the end of each month, serum concentrations of follicle-stimulating hormone (FSH), testosterone, inhibin B, and sex hormone binding globulin (SHBG) were measured. At baseline and at the end, semen analysis was performed and sperm concentration, spermatozoal motility and normal sperm forms were determined. MAIN OUTCOME MEASURE(S): Gonadotropin, testosterone, inhibin-B levels, total sperm count, sperm morphology and motility. RESULT(S): Toremifene administration resulted in a significant increase in FSH, testosterone, SHBG, and inhibin B levels, as well as in sperm concentration, percentage motility and normal sperm forms. Twenty-two men's partners achieved pregnancy within 2 months of the end of treatment. At the end of the third month, serum FSH levels were significantly higher in the men whose partners did not achieve pregnancy, and total sperm count and normal sperm forms were significantly lower compared with the group of men whose partners achieved pregnancy. CONCLUSION(S): Toremifene administration for a period of 3 months in men with idiopathic oligozoospermia is associated with significant improvements of sperm count, motility, and morphology, mediated by increased gonadotropin secretion and possibly a direct beneficial effect of toremifene on the testes. The above findings are also indicative of a better testicular exocrine (improved sperm parameters) response to treatment in men whose partners achieved pregnancy compared with those who did not. Further randomized, placebo-controlled trials should be conducted to determine whether this particular selective estrogen receptor modulator can be useful as an initial approach in men with oligozoospermia. PMID- 17412337 TI - Arg72Pro p53 polymorphism in Italian women: no association with endometriosis. AB - p53 codon 72 polymorphism in Italian women have a minor role in determining genetic susceptibility to endometriosis. The racial differences, in association with other risk factors, might be underlined in endometriotic disease. PMID- 17412338 TI - Single large cystic adenomyoma of the uterus after cornual pregnancy and curettage. AB - A case of single large cystic adenomyoma of the uterus (anechoic area 1.6 cm in diameter) was diagnosed by surgery and histopathologic analysis more than 3 years after a transcervical curettage for an early right-cornual pregnancy. PMID- 17412339 TI - Role of embryo transfer in fellowship training. AB - Our objective was to survey training in ET techniques among fellows, its perceived importance, and potential barriers to ET training during fellowship. Although ET training remains an important issue for most fellows and recent graduates, 44% of respondents did not receive this training during their fellowship. PMID- 17412340 TI - Outcome of in vitro fertilization treatment in infertile women conservatively treated for endometrial adenocarcinoma. AB - OBJECTIVE: To evaluate IVF characteristics and outcome of infertile women conservatively treated for well-differentiated endometrial adenocarcinoma. DESIGN: Observational case series study. SETTING: The Department of Gynecology Oncology and IVF unit of Chaim Sheba Medical center, Tel-Hashomer, Israel. PATIENT(S): Eight women referred to IVF treatment because of failure of other fertility treatments after conservative treatment of endometrial adenocarcinoma. All women were selected carefully for conservative treatment at the Department of Gynecology Oncology of the Chaim Sheba Medical Center after a thorough metastatic workup. INTERVENTION(S): In vitro fertilization/intracytoplasmic sperm injection treatment. MAIN OUTCOME MEASURE(S): Serum E(2) levels at hCG administration, endometrial thickness, number of oocytes retrieved, fertilization rate, pregnancy and delivery rate. RESULT(S): All women were nulliparous before IVF treatment, and only one woman was older than 35 years. In four of them, endometrial adenocarcinoma was diagnosed during infertility workup or treatment. A total of 31 IVF cycles were performed. The mean number of oocytes retrieved was 9.4 (95% confidence interval, 5.1-13.6), and fertilization rate was 58.6%. Six women (75%) conceived, and four (50%) delivered six healthy offspring. CONCLUSION(S): In vitro fertilization treatment of infertile women conservatively treated for well differentiated endometrial adenocarcinoma is highly successful and offers the opportunity to cryopreserve embryos for future use. PMID- 17412341 TI - Functional analysis of Nkx2.1 and Pax9 for calcitonin gene transcription. AB - Nkx2.1 (TTF-1), a homeodomain-containing transcription factor essential for specific gene expression in thyroid follicular cells, exists also in the thyroidal C cells that secrete calcitonin (CT). In this report, we examined the effect of Nkx2.1 on the CT gene transcription. Luciferase reporter assay using the 2kbp promoter sequence of rat CT/CGRP gene revealed that Nkx2.1 induced a significant increase in the promoter transcription. Furthermore, we detected Pax1 and/or Pax9 gene expression in mammalian medullary thyroid carcinoma cell lines, rat rMTC and human TT cells, and in mammalian thyroid glands by RT-PCR. The Pax9 mRNA, expressed in the TT cells and rat thyroid, was then isolated by cDNA cloning. Sequence analysis showed that both rat and human Pax9 proteins contained characteristic domains: i.e. the paired domain and octapeptide motif. Alternative transcripts encoding Pax9 isoforms were not identified in the rat thyroid or TT cells. Dual luciferase assay indicated that Pax9 did not increase transcription from the CT/CGRP promoter. Pax9 also showed no cooperative effects when it was co transfected with Nkx2.1. The results suggest that CT gene expression could be directly activated by Nkx2.1, whereas Pax9 is not involved in transcription from the 2kbp CT promoter. PMID- 17412342 TI - Non-invasive measurement of 11-ketotestosterone, cortisol and androstenedione in male three-spined stickleback (Gasterosteus aculeatus). AB - The androgen 11-ketotestosterone (11-KT) plays an important role in reproductive physiology and behaviour in male teleosts. In the three-spined stickleback, Gasterosteus aculeatus, the plasma concentrations of 11-KT are related to the breeding status of the fish. Sticklebacks are relatively small (generally less than 1g) and in order to obtain sufficient plasma for assay of 11-KT, it has been necessary in the past to sacrifice the fish. In this paper, we report on the development of a non-invasive procedure for measuring 11-KT, cortisol and androstenedione (Ad) in the three-spined stickleback. Validation of the procedure included the demonstration that the rate of release of steroids into the water was correlated to their plasma concentrations. Ten males that were kept at a low temperature and short photoperiod were moved to high temperature and long photoperiod to initiate spermatogenesis and breeding. Every two to four days, for a total of 53 days, males were removed and placed in a beaker containing 50-ml water for 30 min. The water was then processed by solid phase extraction for radioimmunoassay. Males were presented with females on days 13/14, 18/19 and 44/45. 11-KT was originally undetectable but built up gradually to reach an average release rate of between 1 and 2.5 ng/g/h between days 16 and 45 and then started to decline (but non-significantly). Ad release reached a plateau of 1 ng/g/h about day 20. However, from days 44/45 to 51, there was a highly significant rise in the rate of release of Ad to 5 ng/g/h. On days 44/45, five of the males mated successfully and five did not. However, there were no significant differences in 11-KT or Ad release rates between the two groups. Cortisol release rates fluctuated with no pattern throughout the study. The results show that it is possible to make measurements on sex and stress steroid production in sticklebacks without recourse to anaesthesia, bleeding or sacrificing the fish. The procedure is potentially a powerful tool for the study of the link between steroids and behaviour in this useful sentinel species. PMID- 17412343 TI - Coccygectomy for coccygodynia: do we really have to wait? AB - AIM: To determine the results of 21 cases of persistent coccygodynia unresponsive to conservative management and treated with coccygectomy. METHODS: Of 81 patients with coccygodynia, 21 underwent surgical excision of the coccyx with a minimum 5 days of antibiotic prophylaxis. All 21 patients were followed for at least 2 years and questioned about their satisfaction with the operation and its timing. Pain levels were recorded preoperatively and during the postoperative period using the VAS scale. RESULTS: The mean VAS score was 51.88 (40-70), and this decreased to 3.17, 2.94 and 2.76 in the 6th, 12th and 24th months, respectively. This change was statistically significant. Of the 21 cases, 17 had an excellent result and 4 had a good result. None had a wound healing problem or infection. All were satisfied with the operation, and all stated that they would have liked to have undergone it sooner. CONCLUSION: For unstable coccygeal fracture and persistent coccygodynia, coccygectomy is a reliable method of treatment with a high satisfaction and a low complication rate. PMID- 17412344 TI - Insulin attenuates vascular smooth muscle calcification but increases vascular smooth muscle cell phosphate transport. AB - Medial artery vascular smooth muscle cell (VSMC) calcification increases the risk of cardiovascular mortality in type 2 diabetes. However, the influence of insulin on VSMC calcification is unclear. We explored the effects of insulin on rat VSMC calcification in vitro and found that in a dose-dependent fashion, insulin attenuates VSMC calcification induced by high phosphate conditions as quantified by the o-cresolphthalein calcium (OCPC) method. In an in vitro model of insulin resistance in which cells are exposed to elevated insulin concentrations and the PI 3-kinase pathway is selectively inhibited, increased VSMC calcification was observed, suggesting that the PI 3-kinase pathway is involved in this attenuating effect of insulin. We postulated that insulin may also have an effect on phosphate or calcium transport in VSMC. We found that insulin increases phosphate transport at 3 and 24 h. This effect was mediated by increased Vmax for phosphate transport but not Km. Because type III sodium-phosphate co-transporters Pit-1 and Pit-2 are found in VSMC, we examined their expression by Western blot and real time RT-PCR. Insulin stimulates Pit-1 mRNA modestly (*p<0.01 versus control), an effect inhibited by PD98059 but not by wortmannin. Pit-1 protein expression is induced by insulin, an effect also inhibited by PD98059 (*p<0.001 versus insulin alone). Our results suggest a role for insulin in attenuating VSMC calcification which may be disrupted in selective insulin signaling impairment seen in insulin resistance. This effect of insulin contrasts with its effect to induce phosphate transport in VSMC. PMID- 17412345 TI - Serum magnesium and lipids: More clarity is needed. PMID- 17412346 TI - Atheroprotective role of interleukin-6 in diet- and/or pathogen-associated atherosclerosis using an ApoE heterozygote murine model. AB - BACKGROUND: Pathogens have been implicated in the pathogenesis of inflammatory atherosclerosis. Given the pleiotropic role of interleukin-6 in the regulation of cytokines, lipid homeostasis, vascular remodeling, and apoptosis we hypothesized that IL-6 plays an important role in development and progression to inflammatory atherosclerosis. METHODS AND RESULTS: To explore the role of IL-6 in inflammation and infection-associated atherosclerosis, 10-week-old ApoE(+/-)-IL-6(+/-) and ApoE(+/-)-IL-6(-/-) mice fed either high fat diet or regular chow diet were inoculated intravenously, once per week for 14 or 24 consecutive weeks with 50 microl live Porphyromonas gingivalis (P.g.) (10(7)CFU) or vehicle (normal saline). Animals were euthanized at 24 weeks of age (14 weeks injection) or 34 weeks of age (24 weeks injection). Histomorphometric analysis of atheromatous lesions, en face analysis over the aortic tree, immunohistochemistry for macrophages and smooth muscle cell, TUNEL staining for apoptotic cells, serum amyloid A (SAA) levels, serum lipids and glucose level, serum cytokines were obtained. ApoE(+/-)-IL-6(-/-) mice showed a significant increase in atheromatous lesions in proximal aorta and aortic tree compared to ApoE(+/-)-IL-6(+/-) mice for all conditions (chow diet and P.g.-inoculated, high fat diet and P.g. inoculated, high fat diet and vehicle-inoculated) at 14 weeks and greater at 24 weeks. SAA levels from ApoE(+/-)-IL-6(-/-) mice were significantly higher than ApoE(+/-)-IL-6(+/-) mice. IL-6 deficiency led to profound changes in plaque composition evidenced by increased macrophage infiltration, apoptosis, lipid content and decreased smooth muscle cell mass reflecting an unstable plaque phenotype. Array analysis revealed increased levels of proinflammatory cytokines in ApoE(+/-)-IL-6(-/-) mice compared to ApoE(+/-)-IL-6(+/-) mice, irrespective of diet or inoculation. CONCLUSION: The genetic deficiency of IL-6 was found to enhance the formation of diet- and/or pathogen-associated atherosclerotic plaques and suggests that IL-6 may play an atheroprotective role. PMID- 17412347 TI - Ethnic differences in the relationship of carotid atherosclerosis to coronary calcification: the Multi-Ethnic Study of Atherosclerosis. AB - Ethnic differences in non-invasive measures of atherosclerosis are increasingly being reported, but the relationship of these measures to each other has not been widely explored. Carotid ultrasonographic and computed cardiac tomographic findings were compared in 6814 participants of White, Black, Hispanic, and Chinese ethnicities free of overt cardiovascular disease. Coronary calcium and carotid atherosclerosis were strongly related to each other in all ethnic groups. Associations of coronary calcium prevalence and common carotid intimal-medial thickness (IMT) differed by ethnicity in women, being weakest among Black women (0.07 mm IMT difference between those with and without coronary calcium) compared to the other three groups (0.10-0.12 mm difference, p=0.007). Estimated percent increments in internal carotid IMT per 10% increment in coronary calcium score were highest in Hispanics (18.5%) and lowest in Blacks (6.1%, p<0.01). Coronary calcium may be less strongly associated with carotid atherosclerosis in Blacks, particularly Black women, than in other ethnic groups. These differences should be pursued for relationships to coronary events to determine whether coronary calcium carries the same risk information in other ethnic groups as it does in Whites. PMID- 17412348 TI - Effect of fenofibrate therapy on paraoxonase1 status in patients with low HDL-C levels. AB - OBJECTIVES: We evaluated the effect of micro-coated fenofibrate on lipid parameters, high sensitivity C-reactive protein and paraoxonase1 levels in dyslipidemic patients with low high-density lipoproteins levels. In addition, the effects of the paraoxonase1 polymorphisms on lipid and paraoxonase1 responses to fenofibrate therapy were examined. METHODS: A total of 61 dyslipidemic patients with low high-density lipoproteins levels were recruited into this study to receive micro-coated fenofibrate (160 mg/day) for 12 weeks. Lipid parameters, C reactive protein, paraoxonase1 concentration and activity were measured at baseline and after 6 and 12 weeks of fenofibrate treatment. Four polymorphisms in both the coding (L55M and Q192R) and regulatory regions (T-108C and G-909C) of human paraoxonase1 were also quantified. RESULTS: Micro-coated fenofibrate significantly decreased total cholesterol, triglycerides, non-high-density lipoprotein cholesterol, oxidized low-density lipoprotein and apolipoprotein-B levels after 6 and 12 weeks (all p<0.001). While high-density lipoprotein and apolipoprotein AI levels were significantly increased by 14.7% and 6.9%, respectively, after 6 weeks and by 17.3% and 7.2%, respectively, after 12 weeks (all p<0.01). There were no significant differences in the mean of low-density lipoprotein and C-reactive protein after fenofibrate treatment. There were significant increases in paraoxonase1 concentration and activity by 7.7% and 5.7% after 6 weeks and by 14.6% and 10.1% after 12 weeks, respectively (all p<0.01). After micro-coated fenofibrate therapy, a significantly positive correlation between the change in high-density lipoprotein and the changes in paraoxonase1 concentration and activity was observed (p=0.001). On the other hand, the changes in paraoxonase1 activity were significantly and negatively correlated with the changes in triglycerides (p=0.007). The therapeutic response of lipid parameters to micro-coated fenofibrate was independent of paraoxonase1 polymorphisms. However, paraoxonase1 Q192R and T-108C polymorphisms significantly affected the increase in paraoxonase1 activity (the highest increase in 192QQ and -108TT) and paraoxonase1 concentration (the highest increase in -108TT). CONCLUSION: Lipid modifying therapy with micro-coated fenofibrate in patients with low high-density lipoprotein levels not only reduced atherogenic lipids (total cholesterol, triglycerides, oxidized low-density lipoprotein and apolipoprotein-B) and increased atheroprotective lipids but also increased paraoxonase1 concentration and activity. Increasing paraoxonase1 levels by fenofibrate may play an important role in decreasing low-density lipoprotein oxidation. PMID- 17412349 TI - The effect of recovery time and test conditions on viscoelastic measures of tensile damage in cortical bone. AB - Stiffness degradation and strength degradation are often measured to monitor and characterize the effects of damage accumulation in bone. Based on evidence that these properties could be affected by not only damage magnitude but also test conditions, the present study investigated the effect of hold condition and recovery time on measures of tensile damage. Machined human femoral cortical bone specimens were subjected to tensile tests consisting of a pre-damage diagnostic loading cycle, a damage loading cycle and post-damage cycle. Controlled variables were recovery time (1, 10, and 100 min) and hold condition (zero load or zero strain) after the damage cycle. Damage measures were calculated as the ratio of each post-damage cycle to the pre-damage value for loading modulus, secant modulus, unloading modulus, stress relaxation and strain (stress) recovery at 1 min post-diagnostic time. The damage cycle caused reductions in all measures, and some measures varied with recovery time and hold condition. Apparent modulus degradation for both hold conditions decreased with recovery time. Stress relaxation was unaffected by recovery time for both hold conditions. Zero-strain hold conditions resulted in lower values for degradation of modulus and change of relaxation. Stress or strain recovery after the damage cycle was evident through 100 min, but 90% of the recovery occurred within 10 min. The results demonstrate that choice of test conditions can influence the apparent magnitude of damage effects. They also indicate that 10 min recovery time was sufficient to stabilize most measures of the damage state. PMID- 17412350 TI - Analysis of new counter-current chromatography operating modes. AB - Some novel cyclic operating modes of counter-current chromatography have been suggested. Chromatographic separation processes with two zones of different partition coefficients and with extraction and scrubbing (washing) zones are mathematically described using the eluting counter-current distribution approach. Two possible cyclic modes of dual counter-current chromatography operation, with simultaneous and alternate transfers of the phases, are analyzed. It is shown that the operating mode with the alternate transfers of the phases provides considerably higher separation efficiency than the mode with simultaneous transfers of both phases. PMID- 17412351 TI - Selective sample treatment using molecularly imprinted polymers. AB - The molecularly imprinted polymers (MIPs) are synthetic polymers possessing specific cavities designed for a target molecule. By a mechanism of molecular recognition, the MIPs are used as selective sorbents for the solid-phase extraction of target analytes from complex matrices. MIPs are often called synthetic antibodies in comparison with immuno-based sorbents; they offer some advantages including easy, cheap and rapid preparation and high thermal and chemical stability. This review describes the use of MIPs in solid-phase extraction with emphasis on their synthesis, the various parameters affecting the selectivity of the extraction, their potential to selectively extract analytes from complex aqueous samples or organic extracts, their on-line coupling with LC and their potential in miniaturized devices. PMID- 17412352 TI - Effect of temperature on gradient reequilibration in reversed-phase liquid chromatography. AB - The effect of mobile phase modifier and temperature on gradient reequilibration is examined using three different stationary phases. The stationary phases studied are a traditional C18 phase, a polar endcapped C18 phase, and an alkyl phase with a polar embedded group. It was observed that both temperature and choice of mobile phase organic modifier had an effect on gradient reequilibration volume on both the traditional C18 stationary phase and the polar endcapped phase. On both these phases, at any given temperature, the reequilibration volume was generally smaller when methanol was used as the mobile phase modifier as compared to acetonitrile. As the temperature is increased from 10 to 50 degrees C, significant reductions in reequilibration volume were observed with both mobile phase modifiers. In contrast, neither temperature nor choice of modifier appeared to have much effect on reequilibration volume when the polar embedded group stationary phase was considered. PMID- 17412353 TI - On the 3-dimensional effects in etched chips for high performance liquid chromatography-separations. AB - In an attempt to quantify the potential of photolithographically etched micro pillar arrays as a perfectly ordered alternative for the packed bed of spheres, the additional band broadening originating from the top and bottom plate has been investigated using computational fluid dynamics simulations. These calculations provide insight in the theoretical expectations that can be made for the experimental work that is currently being conducted by a number of groups. The calculations show that the additional band broadening contribution can be expected to go through a transient regime as a function of the axial distance along the array. In its fully developed regime and in the most relevant velocity range, the top and bottom wall contribution almost doubles the band broadening compared to the band broadening in a perfectly ordered array of non-porous, non retentive pillars without top and bottom wall. Compared to the band broadening in an array of porous, retentive pillars on the other hand, the top and bottom wall effect can be expected to become negligible. A simplified, phenomenological model yielding a first principles prediction of both the transient and the steady-state top and bottom wall band broadening as a function of the inter-pillar distance and the pillar height is proposed and shows good qualitative agreement with the exact calculations. PMID- 17412354 TI - An anion-exchange high-performance liquid chromatography method coupled to total organic carbon determination for the analysis of water-soluble organic aerosols. AB - The chemical composition of water-soluble organic carbon (WSOC) in atmospheric aerosol particles is largely unexplored, due to the myriad of individual compounds, which has hampered attempts to attain a full characterization at the molecular level. An alternative approach, focusing on the analysis of a few main chemical classes, allowed the quantitative fractionation of WSOC into neutral compounds (NC), mono- and di-acids (MDA) and polyacids (PA) through an anion exchange liquid chromatographic method. Previous attempts to quantify NC, MDA and PA relied on a low-pressure chromatographic technique using a volatile buffer, followed by total organic carbon (TOC) analysis of the fractions, or alternatively on a faster HPLC-UV method which provided a quantification of the fractions based on empirical relationships between UV signal and TOC concentration. Here, we report an upgraded anion-exchange HPLC technique, allowing direct TOC analysis of the eluted fractions, without any pre-treatment, thus permitting a great simplification of quantitative analysis and preventing sample losses. The new HPLC-TOC methodology, employing completely inorganic eluents shows the same efficiency of the former HPLC-UV method employing organic additives, with the exception of phenolic compounds, which are retained on the column by secondary interactions, and low-molecular weight tricarboxylic acids, which are recovered among MDA. The new anion-exchange liquid chromatographic method can recover a substantial amount (86-100%) of water-soluble organic carbon from atmospheric aerosol extracts, thus allowing a higher retention and fractionation capacity with respect to alternative techniques, like solid phase extraction. PMID- 17412355 TI - In-capillary derivatization and laser-induced fluorescence detection for the analysis of organophosphorus pesticides by micellar electrokinetic chromatography. AB - We developed a rapid and sensitive method using in-capillary derivatization and laser-induced fluorescence (LIF) detection for the fully automated analysis of organophosphorus pesticides (OPPs), including glufosinate, aminomethylphosphonic acid (AMPA) and glyphosate by micellar electrokinetic chromatography (MEKC). The potential of 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) as in-capillary derivatization reagent is described for the first time. The unique feature of this MEKC method is the capillary being used as a small reaction chamber. In in capillary derivatization, the sample and reagent solutions were injected directly into the capillary by tandem mode, followed by an electrokinetic step to enhance the mixing efficiency of analytes and reagent plugs in accordance with their different electrophoretic mobilities. Standing a specified time for reaction, the derivatives were then immediately separated and determined. Careful optimization of the derivatization and separation conditions allowed the determination of glufosinate, AMPA and glyphosate with detection limits of 2.8, 3.6 and 32.2 ng/mL, respectively. These detection limits were comparable to those of 1.4, 1.9 and 23.8 ng/mL obtained from conventional pre-capillary derivatization. Furthermore, repeatability better than 0.40% for migration time and 3.4% for peak area, as well as shorter migration time, was obtained. The method was successfully applied to the analysis of spiked river water sample with satisfactory results. PMID- 17412356 TI - Wormlike micelles in Tween-80/CmEO3 mixed nonionic surfactant systems in aqueous media. AB - Formation and rheological behaviour of viscoelastic wormlike micellar solution in aqueous mixed system of nonionic surfactants, polyoxyethylene sorbitan monooleate (Tween-80) and trioxyethylene alkyl ether (C(m)EO(3), m=12, 14, and 16) was studied. The semi-dilute aqueous solution of Tween-80, in presence of C(m)EO(3) shows a sharp increase in viscosity leading to the formation of a gel-like highly viscoelastic solution reaching a maximum, but decreases beyond an intermediate concentration and finally phase separates. When C(12)EO(3) is replaced by C(14)EO(3) the micellar growth occurs more effectively. However, with further increase in alkyl chain length, i.e. with C(16)EO(3), phase separation occurs before a viscoelastic solution is formed. The effect of temperature on the water/Tween-80/C(14)EO(3) mixed surfactant system was also studied. With increase in temperature, viscosity increases more promptly and the viscosity maximum is attained at relatively lower concentration of C(14)EO(3). Enhanced one dimensional micellar growth with increase in the temperature is mainly attributed to the decrease in the spontaneous curvature of the aggregates. Structural investigation by means of small-angle X-ray scattering (SAXS) technique of micelles in aqueous solution of Tween-80 and mixed water/Tween-80/C(m)EO(3) systems confirmed the one dimensional micellar growth in the above system. PMID- 17412357 TI - Enantioselective crystallization of histidine on chiral self-assembled films of cysteine. AB - In this paper, the preparation and use of chiral surfaces derived from enantiomerically pure crystals of amino acids are described. For this purpose, a self-assembly process to grow thin chiral films of (+)-L- or (-)-D-cysteine on gold surfaces was chosen. These chiral films were utilized as crystallization catalysts in the crystallization of enantiomers from solutions. To demonstrate the chiral discrimination power of the chiral surfaces in crystallization processes, the crystallization of racemic histidine onto the chiral films was investigated. Our study demonstrates the potential application of chiral films to control chirality throughout crystallization, where one enantiomer crystallizes onto the chiral surfaces with relative high enantiomeric excess. In addition, crystallization of pure histidine enantiomers onto chiral films results in strong crystal morphology modification with preferred orientation. PMID- 17412358 TI - Modeling cold tolerance in the mountain pine beetle, Dendroctonus ponderosae. AB - Cold-induced mortality is a key factor driving mountain pine beetle, Dendroctonus ponderosae, population dynamics. In this species, the supercooling point (SCP) is representative of mortality induced by acute cold exposure. Mountain pine beetle SCP and associated cold-induced mortality fluctuate throughout a generation, with the highest SCPs prior to and following winter. Using observed SCPs of field collected D. ponderosae larvae throughout the developmental season and associated phloem temperatures, we developed a mechanistic model that describes the SCP distribution of a population as a function of daily changes in the temperature dependent processes leading to gain and loss of cold tolerance. It is based on the changing proportion of individuals in three states: (1) a non cold-hardened, feeding state, (2) an intermediate state in which insects have ceased feeding, voided their gut content and eliminated as many ice-nucleating agents as possible from the body, and (3) a fully cold-hardened state where insects have accumulated a maximum concentration of cryoprotectants (e.g. glycerol). Shifts in the proportion of individuals in each state occur in response to the driving variables influencing the opposite rates of gain and loss of cold hardening. The level of cold-induced mortality predicted by the model and its relation to extreme winter temperature is in good agreement with a range of field and laboratory observations. Our model predicts that cold tolerance of D. ponderosae varies within a season, among seasons, and among geographic locations depending on local climate. This variability is an emergent property of the model, and has important implications for understanding the insect's response to seasonal fluctuations in temperature, as well as population response to climate change. Because cold-induced mortality is but one of several major influences of climate on D. ponderosae population dynamics, we suggest that this model be integrated with others simulating the insect's biology. PMID- 17412359 TI - Phenology, distribution, and host specificity of Solenopsis invicta virus-1. AB - Studies were conducted to examine the phenology, geographic distribution, and host specificity of the Solenopsis invicta virus-1 (SINV-1). Two genotypes examined, SINV-1 and -1A, exhibited similar seasonal prevalence patterns. Infection rates among colonies of S. invicta in Gainesville, Florida, were lowest from early winter (December) to early spring (April) increasing rapidly in late spring (May) and remaining high through August before declining again in the fall (September/October). Correlation analysis revealed a significant relationship between mean monthly temperature and SINV-1 (p<0.0005, r=0.82) and SINV-1A (p<0.0001, r=0.86) infection rates in S. invicta colonies. SINV-1 was widely distributed among S. invicta populations. The virus was detected in S. invicta from Argentina and from all U.S. states examined, with the exception of New Mexico. SINV-1 and -1A were also detected in other Solenopsis species. SINV-1 was detected in Solenopsis richteri and the S. invicta/richteri hybrid collected from northern Alabama and Solenopsis geminata from Florida. SINV-1A was detected in S. geminata and Solenopsis carolinensis in Florida and the S. invicta/richteri hybrid in Alabama. Of the 1989 arthropods collected from 6 pitfall trap experiments from Gainesville and Williston, Florida, none except S. invicta tested positive for SINV-1 or SINV-1A. SINV-1 did not appear to infect or replicate within Sf9 or Dm-2 cells in vitro. The number of SINV-1 genome copies did not significantly increase over the course of the experiment, nor were any cytopathic effects observed. Phylogenetic analyses of SINV-1/-1A nucleotide sequences indicated significant divergence between viruses collected from Argentina and the U.S. PMID- 17412360 TI - Sydney rock oyster (Saccostrea glomerata) hemocytes: morphology and function. AB - In this study, three major hemocyte types were identified in the Sydney rock oyster. They were characterized primarily by light and electron microscopy based on the presence or absence of granules and nucleus to cytoplasm ratios. Hemoblast like cells were the smallest cell type 4.0+/-0.4microm and comprised 15+/-3% of the hemocyte population. They had large nuclei and scanty basic cytoplasm. This cell type also had some endoplasmic reticuli and mitochondria. The second major type were hyalinocytes. Hyalinocytes represented 46+/-6% of all hemocytes. They were large cells (7.1+/-1.0microm) that had low nucleus:cytoplasm ratios and agranular basic or acidic cytoplasm. Hyalinocytes had the ability to phagocytose yeast cells and formed the core of hemocyte aggregates associated with agglutination. Four discrete sub-populations of hyalinocytes were identified. The third major cell type were the granulocytes, comprising 38+/-1% of the hemocyte population. These cells were large (9.3+/-0.3microm) and were characterized by cytoplasm containing many acidic or basic granules. Granulocytes were more phagocytic than hyalinocytes and they formed the inner layer of hemocytes during the encapsulation of fungal hyphae. Five discrete sub-populations of granulocytes were identified based on the types of granules in their cytoplasm. Flow cytometry showed that the hemocytes of rock oysters could be divided into between two and four major cell types based on their light scattering properties. The most common of the cell types identified by flow cytometry corresponded to hyalinocytes and granulocytes. Cytochemical assays showed that most enzymes associated with immunological activity were localized in granulocytes. Their granules contained acid phosphatase, peroxidase, phenoloxidase, superoxide and melanin. Hyalinocytes were positive only for acid phosphatase. All of these observations suggest that Sydney rock oysters have a broad variety of functionally specialized hemocytes, many of which are involved in host defense. PMID- 17412361 TI - Thermodynamics of the coil <==> beta-sheet transition in a membrane environment. AB - Biologically important peptides such as the Alzheimer peptide Abeta(1-40) display a reversible random coil <==>beta-structure transition at anionic membrane surfaces. In contrast to the well-studied random coil left arrow over right arrow alpha-helix transition of amphipathic peptides, there is a dearth on information on the thermodynamic and kinetic parameters of the random coil left arrow over right arrow beta-structure transition. Here, we present a new method to quantitatively analyze the thermodynamic parameters of the membrane-induced beta structure formation. We have used the model peptide (KIGAKI)(3) and eight analogues in which two adjacent amino acids were substituted by their d enantiomers. The positions of the d,d pairs were shifted systematically along the three identical segments of the peptide chain. The beta-structure content of the peptides was measured in solution and when bound to anionic lipid membranes with circular dichroism spectroscopy. The thermodynamic binding parameters were determined with isothermal titration calorimetry and the binding isotherms were analysed by combining a surface partition equilibrium with the Gouy-Chapman theory. The thermodynamic parameters were found to be linearly correlated with the extent of beta-structure formation. beta-Structure formation at the membrane surface is characterized by an enthalpy change of DeltaH(beta)=-0.23 kcal/mol per residue, an entropy change of DeltaS(beta)=-0.24 cal/mol K residue and a free energy change of DeltaG(beta)=-0.15 kcal/mol residue. An increase in temperature induces an unfolding of beta-structure. The residual free energy of membrane induced beta-structure formation is close to that of membrane-induced alpha-helix formation. PMID- 17412362 TI - Relationships of DEX/CRH and GHRH test results to the outcome of depression- preliminary results suggest the GHRH test may predict relapse after discharge. AB - To explore and compare hypothalamic-pituitary-somatotropic (HPS) axis function and hypothalamic-pituitary-adrenocortical (HPA) axis function in depression, the dexamethasone (DEX)/CRH test and growth hormone releasing hormone (GHRH) test were prospectively performed on patients with depression at the time of admission and discharge. The patients who relapsed within six months after discharge exhibited significantly lower growth hormone (GH) responses to GHRH at the time of discharge than those who did not relapse. There were no significant correlations between GH response to GHRH and the results of DEX/CRH tests after controlling for age, sex, and body mass index. The findings of this study suggest that results of the GHRH test may be a predictor of future relapse in patients with depression. PMID- 17412363 TI - Effects of haloperidol and risperidone on cerebrohemodynamics in drug-naive schizophrenic patients. AB - BACKGROUND: Use of antipsychotics may be associated with cerebrovascular adverse events in psychotic patients. In this study, the effects of haloperidol and risperidone on the cerebral hemodynamics and the possible relationships between antipsychotics and cerebrovascular risks tendency were evaluated by Transcranial Doppler ultrasonography (TCD). METHODS: Twenty drug-nai ve schizophrenic patients and 20 normal control subjects were included. The patients were divided into haloperidol- and risperidone-treated groups and received treatment for 8 weeks double-blindly. The subjects' cerebral blood flow mean velocities (MV) and pulsatility index (PI) were measured weekly by TCD. The Positive and Negative Syndrome Scale for schizophrenia (PANSS) was used to assess the patients' psychopathological symptoms. RESULTS: Increased MV and decreased PI were found significantly in drug-nai ve schizophrenic patients than normal subjects before treatment (p<0.01). The decreased PI could be normalized after 8 weeks of antipsychotic treatment, while the increased MV could not. Treatment with haloperidol could significantly increase the PI than the treatment with risperidone (p<0.01) throughout the treatment course. The PANSS scores of both groups were significantly improved (p<0.05) at the endpoints of treatment. CONCLUSIONS: Our findings indicate that haloperidol may affect the cerebral hemodynamics in drug-naive schizophrenics more prominently than that of risperidone via TCD monitoring. PMID- 17412364 TI - The role of HLA-DRB1 alleles on susceptibility and outcome of a Portuguese Multiple Sclerosis population. AB - BACKGROUND: The association between susceptibility to multiple sclerosis (MS) and HLA-DRB1*15 has been reported in various European populations. OBJECTIVE: To investigate the relationship between MS, HLA-DRB1*15 and other DRB1 alleles in a Portuguese population and their association with clinical course of MS. METHODS: The HLA-DRB1 alleles were analyzed by PCR-SSP in 248 MS patients and 282 healthy controls. In order to relate HLA-DRB1 alleles to disease aggressiveness, patients with relapsing remitting MS and secondary progressive MS were subdivided into 3 groups: 'benign' MS patients who maintain an Extended Disability Status Scale (EDSS) score of 3 after the same period and 'aggressive' MS those with EDSS>or=6 within 15 years of disease onset. RESULTS: As expected, a higher frequency of HLA-DRB1*15 was found in MS patients (29.8% vs 19.9%, odds ratio (OR)=1.72, 95% CI=1.15-2.56, p=0.008). The HLA-DRB1*03 allele was positively associated with MS in the overall patient population (22.6% vs 15.6%, OR=1.58, 95% CI=1.02-2.45). Concerning disease aggressiveness, HLA-DRB1*15 occurred more frequently in the group with benign disease (42.6% vs 19.9%, OR=2.99, 95% CI=1.56 5.72) and in the group with non-benign disease (34.1% vs 19.9%, OR=2.09, 95% CI=1.05-4.16) compared with controls. When time to reach an EDSS=3 or EDSS=6 was considered as end point, HLA-DRB1*15 negative patients were found to have a worse prognosis. CONCLUSIONS: In this population of Portuguese MS patients, the HLA DRB1*15 allele is established as a genetic marker for susceptibility to MS and is also associated with a better outcome. PMID- 17412365 TI - How regulatory CD25(+)CD4(+) T cells impinge on tumor immunobiology? On the existence of two alternative dynamical classes of tumors. AB - Aiming to get a better insight on the impact of regulatory CD25(+)CD4(+) T cells in tumor immunobiology, a simple mathematical model was formulated and studied. This model is an extension of a previous model for the dynamics of autoreactive regulatory cells and effector cells that interact upon their co-localized activation at the antigen presenting cells (APCs). It assumes that tumor growth stimulates the activation and migration to the adjacent lymph node of fresh APCs loaded with tumor antigens. These APCs stimulate the growth of both effector and regulatory T cells, which may then migrate to the tumor site and induce tumor cell destruction. Our results predict the existence of two alternative dynamic modes of unbounded tumor growth. In the first mode, the tumor induces the expansion of effector T cells that outcompete regulatory T cells, but nevertheless fail to control the tumor. In the second mode, the tumor induces a balanced expansion of both effector and regulatory T cells, which prevents the tumor from being destroyed by the immune cells. Tumors characterized by a high specific growth rate, low immunogenicity, and that are relatively resistant to T cell destructive functions, will grow in the first mode; conversely, tumors that have a slow specific growth rate, that are immunogenic, and/or that are more sensitive to destruction by T cells will grow in the second mode. Overall, this result provides a simple explanation to the fact that the development of some tumors expands regulatory T cells while others do not, predicting how some key dynamical properties of the tumor determine either one or the other type of behavior. PMID- 17412366 TI - Acetylcholine-induced proliferation of fibroblasts and myofibroblasts in vitro is inhibited by tiotropium bromide. AB - Acetylcholine (ACh) has been suggested to exert various pathophysiological activities in the airways in addition to vagally-induced bronchoconstriction. This archetypal neurotransmitter and other components of the cholinergic system are expressed in a number of non-neuronal cells in the airways. Non-neuronal ACh released from these cells may affect fibroblasts (Fb) as well as inflammatory cells in lung tissue. Tiotropium bromide is a once-a-day antimuscarinic drug, marketed under the brand name Spiriva, for the treatment of chronic obstructive pulmonary disease (COPD). Besides its proven direct bronchodilatory activity, recent evidence suggests that tiotropium may be able to reduce the frequency of exacerbations and attenuate the decline in lung function, thus improving the course of obstructive airway diseases. The aim of the present study was to investigate the effects of tiotropium on the ACh-induced proliferation of primary human Fb isolated from biopsies of lung fibrosis patients and myofibroblasts (MyFb) derived from these cells. A human lung Fb cell line acted as control. Expression of muscarinic receptor subtypes M1, M2 and M3 was demonstrated by RT PCR in both cell types. Acetylcholine stimulated proliferation in all cells investigated. Tiotropium concentration-dependently inhibited the ACh-induced proliferation in both the Fb and MyFb with a maximum effect at 30 nM. These results suggest that cholinergic stimuli mediated by muscarinic receptors could contribute to remodeling processes in chronic airway disease. Tiotropium bromide may have a beneficial influence on airway remodeling processes in chronic airway diseases through antiproliferative effects on fibroblasts and myofibroblasts. PMID- 17412367 TI - Blowing-up of deterministic fixed points in stochastic population dynamics. AB - We discuss the stochastic dynamics of biological (and other) populations presenting a limit behaviour for large environments (called deterministic limit) and its relation with the dynamics in the limit. The discussion is circumscribed to linearly stable fixed points of the deterministic dynamics, and it is shown that the cases of extinction and non-extinction equilibriums present different features. Mainly, non-extinction equilibria have associated a region of stochastic instability surrounded by a region of stochastic stability. The instability region does not exist in the case of extinction fixed points, and a linear Lyapunov function can be associated with them. Stochastically sustained oscillations of two subpopulations are also discussed in the case of complex eigenvalues of the stability matrix of the deterministic system. PMID- 17412368 TI - Bringing consistency to simulation of population models--Poisson simulation as a bridge between micro and macro simulation. AB - Population models concern collections of discrete entities such as atoms, cells, humans, animals, etc., where the focus is on the number of entities in a population. Because of the complexity of such models, simulation is usually needed to reproduce their complete dynamic and stochastic behaviour. Two main types of simulation models are used for different purposes, namely micro simulation models, where each individual is described with its particular attributes and behaviour, and macro-simulation models based on stochastic differential equations, where the population is described in aggregated terms by the number of individuals in different states. Consistency between micro- and macro-models is a crucial but often neglected aspect. This paper demonstrates how the Poisson Simulation technique can be used to produce a population macro-model consistent with the corresponding micro-model. This is accomplished by defining Poisson Simulation in strictly mathematical terms as a series of Poisson processes that generate sequences of Poisson distributions with dynamically varying parameters. The method can be applied to any population model. It provides the unique stochastic and dynamic macro-model consistent with a correct micro-model. The paper also presents a general macro form for stochastic and dynamic population models. In an appendix Poisson Simulation is compared with Markov Simulation showing a number of advantages. Especially aggregation into state variables and aggregation of many events per time-step makes Poisson Simulation orders of magnitude faster than Markov Simulation. Furthermore, you can build and execute much larger and more complicated models with Poisson Simulation than is possible with the Markov approach. PMID- 17412369 TI - Lipid peroxidation-derived etheno-DNA adducts in human atherosclerotic lesions. AB - Atherosclerosis and cancer are characterized by uncontrolled cell proliferation and share common risk factors, such as cigarette smoking, dietary habits and ageing. Growth of smooth muscle cells (SMCs) in atherosclerotic plaques may result from DNA damage, caused either by exogenous mutagens or by agents endogenously generated due to oxidative stress and lipid peroxidation (LPO). Hydroxy-2-nonenal (HNE), a major LPO product, binds covalently to cellular DNA to form the exocyclic etheno-DNA-base adducts, 1,N(6)-ethenodeoxyadenine (varepsilondA) and 3,N(4)-ethenodeoxycytosine (varepsilondC). By applying an ultrasensitive (32)P-postlabeling-immunoaffinity method, varepsilondA and varepsilondC were quantified in abdominal aorta SMCs from 13 atherosclerotic patients and 3 non-smoking subjects without atherosclerotic lesions. The levels of etheno-adducts ranged for varepsilondA from 2.3 to 39.6/10(8)dA and for varepsilondC from 10.7 to 157.7/10(8)dC, with a high correlation between varepsilondA and varepsilondC (r=0.84, P=0.0001). Etheno-adduct levels were higher in atherosclerotic smokers than in ex-smokers for both varepsilondA (means 15.2 versus 7.3, P=0.06) and varepsilondC (71.9 versus 51.6, not significant). varepsilondC levels were higher in either ex-smokers (P=0.03) or smokers (P=0.07) than in non-smokers. There was a poor correlation between either varepsilondA or varepsilondC and 8-hydroxy-2'-deoxyguanosine, whereas significant positive correlations were detected with the levels of several postlabeled bulky aromatic DNA adducts. In conclusion, two different types of DNA damage may be involved in atherosclerotic plaque formation and progression: (i) bulky aromatic compounds, to which aorta SMCs are chronically exposed in smokers, can either covalently bind to DNA, induce redox-cycling via quinone intermediates and/or activate local chronic inflammatory processes in the arterial wall; ii) this in turn leads to a self perpetuating generation of reactive oxygen species, LPO-products and increasing DNA-damage, as documented by the presence of high levels of miscoding etheno-DNA adducts in human aorta SMCs. PMID- 17412370 TI - Chromosomal aberrations by fluorescence in situ hybridization (FISH)--Biomarker of exposure to carcinogenic PAHs. AB - The fluorescence in situ hybridization (FISH) technique with whole chromosome painting for chromosomes #1 and #4 was used to study the impact of air pollution containing higher concentrations of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) in three European cities, Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). In each site were followed an exposed group, who were police officers or bus drivers who work usually through busy streets for at least 8h, and a reference group, who spent more than 90% of their daily time indoors. In Prague, a significant increase was observed in percentage of aberrant cells (% AB.C.) in the police officers compared to the reference group (0.33+/-0.25 versus 0.24+/-0.18, p<0.05). In Kosice, the exposed group differed from reference in the endpoints F(G)/100 1.52+/-1.18 versus 1.12+/-1.30, p<0.05; % AB.C. 0.30+/-0.19 versus 0.21+/-0.20, p<0.05; t/1000 3.91+/-3.18 versus 2.84+/-3.10, p<0.05. In Sofia were followed two exposed groups: police officers and bus drivers. All FISH endpoints were significantly higher in police officers compared to reference group (F(G)/100 1.60+/-0.99 versus 0.82+/-0.79, p<0.01; % AB.C. 0.25+/-0.14 versus 0.13+/-0.13, p<0.01; t/1000 4.19+/-2.65 versus 2.13+/-2.05, p<0.05; rcp 1.46+/-1.07 versus 0.70+/-0.76, p<0.05). In bus drivers compared to reference there was an increase in % AB.C. (0.25+/-0.18 versus 0.13+/-0.13, p<0.05). This is the first study when FISH method was used to analyze the impact of environmental air pollution. According to the original hypothesis it is expected that the most important group of chemicals responsible for the biological activity of air pollution represent c-PAHs. PMID- 17412371 TI - PAH-DNA adducts in environmentally exposed population in relation to metabolic and DNA repair gene polymorphisms. AB - Epidemiologic studies indicate that prolonged exposure to particulate air pollution may be associated with increased risk of cardiovascular diseases and cancer in general population. These effects may be attributable to polycyclic aromatic hydrocarbons (PAHs) adsorbed to respirable air particles. It is expected that metabolic and DNA repair gene polymorphisms may modulate individual susceptibility to PAH exposure. This study investigates relationships between exposure to PAHs, polymorphisms of these genes and DNA adducts in group of occupationally exposed policemen (EXP, N=53, males, aged 22-50 years) working outdoors in the downtown area of Prague and in matched "unexposed" controls (CON, N=52). Personal exposure to eight carcinogenic PAHs (c-PAHs) was evaluated by personal samplers during working shift prior to collection of biological samples. Bulky-aromatic DNA adducts were analyzed in lymphocytes by (32)P-postlabeling assay. Polymorphisms of metabolizing (GSTM1, GSTP1, GSTT1, EPHX1, CYP1A1-MspI) and DNA repair (XRCC1, XPD) genes were determined by PCR-based RFLP assays. As potential modifiers and/or cofounders, urinary cotinine levels were analyzed by radioimmunoassay, plasma levels of vitamins A, C, E and folates by HPLC, cholesterol and triglycerides using commercial kits. During the sampling period ambient particulate air pollution was as follows: PM10 32-55microg/m(3), PM2.5 27 38microg/m(3), c-PAHs 18-22ng/m(3); personal exposure to c-PAHs: 9.7ng/m(3) versus 5.8ng/m(3) (P<0.01) for EXP and CON groups, respectively. The total DNA adduct levels did not significantly differ between EXP and CON groups (0.92+/ 0.28adducts/10(8) nucleotides versus 0.82+/-0.23adducts/10(8) nucleotides, P=0.065), whereas the level of the B[a]P-"like" adduct was significantly higher in exposed group (0.122+/-0.036adducts/10(8) nucleotides versus 0.099+/ 0.035adducts/10(8) nucleotides, P=0.003). A significant difference in both the total (P<0.05) and the B[a]P-"like" DNA adducts (P<0.01) between smokers and nonsmokers within both groups was observed. A significant positive association between DNA adduct and cotinine levels (r=0.368, P<0.001) and negative association between DNA adduct and vitamin C levels (r=-0.290, P=0.004) was found. The results of multivariate regression analysis showed smoking, vitamin C, polymorphisms of XPD repair gene in exon 23 and GSTM1 gene as significant predictors for total DNA adduct levels. Exposure to ambient air pollution, smoking, and polymorphisms of XPD repair gene in exon 6 were significant predictors for B[a]P-"like" DNA adduct. To sum up, this study suggests that polymorphisms of DNA repair genes involved in nucleotide excision repair may modify aromatic DNA adduct levels and may be useful biomarkers to identify individuals susceptible to DNA damage resulting from c-PAHs exposure. PMID- 17412372 TI - Tranylcypromine enhancement of nicotine self-administration. AB - Tobacco use has one of the highest rates of addiction of any abused drug. Paradoxically, in animal models, nicotine appears to be a weak reinforcer. We report here that the inhibition of monoamine oxidase (MAO), a major effect of tobacco smoke, increases the reinforcing effect of nicotine. Rats (aged postnatal day 27 and 90) were tested for self-administration, without prior response training, in five daily 3-h sessions. Whereas control rats did not self administer nicotine, low doses of nicotine (2.5 to 21 microg/kg/injection) were avidly self-administered following a pretreatment with tranylcypromine (3 mg/kg), an irreversible and non-selective MAO inhibitor. Tranylcypromine-enhanced nicotine (10 microg/kg/injection, i.v.) self-administration was reduced by systemic injection of a D1-dopaminergic receptor antagonist, SCH23390 (0.02 mg/kg). Moreover, an increase in extracellular dopamine in the nucleus accumbens was detected, using microdialysis, following nicotine (60 microg/kg) injection in tranylcypromine pre-treated rats. Depending on the time of tranylcypromine pretreatment (20 or 1 h), MAO activity was decreased by 72% and 99% and nicotine intake at day 5 was increased by 619 and 997%, respectively. Taken together, these results indicate that in a stringent self-administration acquisition test, MAO inhibition increases the rewarding effect of low doses of nicotine, possibly via a dopamine-dependent mechanism. PMID- 17412373 TI - Mental rotation of body parts and non-corporeal objects in patients with idiopathic cervical dystonia. AB - Mental rotation of body parts is performed through inner simulation of actual movements, and is likely to rely upon cortical and subcortical systems (e.g. motor and premotor areas and basal ganglia) involved in motor planning and execution. Studies indicate that sensory and motor deficits, such as for example pain, limb amputation or focal hand dystonia, bring about a specific impairment in mental rotation of the affected body parts. Here we explored the ability of patients affected by idiopathic cervical dystonia (CD) to mentally rotate affected (neck) and unaffected (hands and feet) body districts. The experimental stimuli consisted of realistic photos of left or right hands or feet and the head of a young men with a black patch on the left or the right eye. As non-corporeal stimulus the front view of a car with a black patch on the left or the right headlight was used. The stimuli were presented at six different degrees of orientations. Twelve CD patients and 12 healthy participants were asked to verbally report whether the hands or feet were left or right, or whether the patch was on the left or the right eye or headlight. Reaction times and accuracy in performing the laterality tasks on the four stimuli were collected. Results showed that CD patients are slow in mental rotation of stimuli representing body parts, namely hand, foot and head. This abnormality was not due to a general impairment in mental rotation per se, since patients' ability to rotate a non corporeal object (a car) was not significantly different from that of healthy participants. We posit that the deficit in mental rotation of body parts in CD patients may derive from a defective integration of body- and world-related knowledge, a process that is likely to allow a general representation of "me in the external world". PMID- 17412374 TI - Yawning in morning and evening types. AB - Yawning occurs more frequently in the early morning and in the late evening, close to sleep onset and after the awakening, and it might be linked to sleep propensity. We aimed to study yawning and its temporal distribution in morning and evening subjects who display different sleep-wake and sleepiness rhythms. Sixteen healthy young adults (8 evening-types and 8 morning-types, matched for age and gender) have been selected and instructed to keep their habitual sleep schedules and to signal every yawning occurrence for three consecutive days. Results show that evening-types yawn more frequently than morning-types, particularly during morning hours. Yawning frequency decreases across daytime in evening-types reaching its lowest level in the early evening and increases thereafter. Instead, in morning-types, yawning frequency remains quite low during daytime and increases in the evening. Moreover, both morning and evening types show a progressive increase of yawning frequency in the hours preceding sleep onset, whereas they differ after the awakening. Evening-types show a higher yawning frequency that remains quite stable in the hours following the awakening, while morning-types display a decline in yawning frequency. Our findings show that the temporal distribution of yawning frequency differs between chronotypes, supporting the hypothesis that differences in sleep-wake rhythm affect yawning, which could represent a behavioural sign of sleep propensity. PMID- 17412375 TI - Enhanced expression of serine proteases during floral senescence in Gladiolus. AB - Programmed cell death during senescence in plants is associated with proteolysis that helps in remobilization of nitrogen to other growing tissues. In this paper, we provide one of the few reports for the expression of specific serine proteases during senescence associated proteolysis in Gladiolus grandiflorus flowers. Senescence in tepals, stamens and carpels results in an increase in total protease activity and a decrease in total protein content. Of the total protease activity, serine proteases account for about 67-70% while cysteine proteases account for only 23-25%. In-gel assays using gelatin as a substrate and specific protease inhibitors reveal the enhanced activity of two trypsin-type serine proteases of sizes 75 kDa and 125 kDa during the course of senescence. The activity of the 125 kDa protease increases not only during tepal senescence but also during stamen and carpel senescence indicating that it is responsive to general senescence signals. PMID- 17412376 TI - Oxylipin formation in Nostoc punctiforme (PCC73102). AB - The dioxygenation of polyunsaturated fatty acids is mainly catalyzed by members of the lipoxygenase enzyme family in flowering plants and mosses. Lipoxygenase products can be metabolized further and are known as signalling substances that play a role in plant development as well as in plant responses to wounding and pathogen attack. Apart from accumulating data in mammals, flowering and non flowering plants, information on the relevance of lipid peroxide metabolism in prokaryotic organisms is scarce. Thus we aimed to isolate and analyze lipoxygenases and oxylipin patterns from cyanobacterial origin. DNA isolated from Nostoc punctiforme strain PCC73102 yielded sequences for at least two different lipoxygenases. These have been cloned as cDNAs and named NpLOX1 and NpLOX2. Both proteins were identified as linoleate 13-lipoxygenases by expression in E. coli. NpLOX1 was characterized in more detail: It showed a broad pH optimum ranging from pH 4.5 to pH 8.5 with a maximum at pH 8.0 and alpha-linolenic acid was the preferred substrate. Bacterial extracts contain more 13-lipoxygenase-derived hydroperoxides in wounded than in non-wounded cells with a 30-fold excess of non esterified over esterified oxylipins. 9-Lipoxygenase-derived derivatives were not detectable. 13-Lipoxygenase-derived hydroperoxides in esterified lipids were present at almost equal amounts compared to non-esterified hydroperoxides in non wounded cells. These results suggest that 13-lipoxygenases acting on free fatty acids dominate in N. punctiforme strain PCC73102 upon wounding. PMID- 17412377 TI - Superantigen-induced cytokine release from whole-blood cell culture as a functional measure of drug efficacy after oral dosing in nonhuman primates. AB - Evaluation of drug efficacy for human diseases is routinely performed in animal models for efficiency and in accordance with FDA regulations. Rhesus macaques have been used as models for various lethal diseases and correlates of immunity, as nonhuman primates (NHP) closely resemble humans. We examined the ex vivo cytokine response of superantigen-stimulated whole-blood cells as a first step to therapeutic efficacy testing for bacterial superantigen-induced shock in NHP after oral dosing of pentoxifylline. Doses of 120mg/kg of pentoxifylline effectively attenuated staphylococcal enterotoxin B-induced tumor necrosis factor alpha (TNFalpha), gamma interferon (IFNgamma) and interleukin 2 (IL-2) in ex vivo culture of NHP whole-blood cells by 88%, 81%, and 76%, respectively, whereas lower doses of 48 or 72mg/kg had no inhibitory effect. Thus cytokine release of stimulated peripheral blood cells provides a convenient biological measurement of the anti-inflammatory potency of pentoxifylline and has the advantage of assessing functional responses to a specific biotoxin of interest. PMID- 17412378 TI - Microsporidiosis in South Africa: PCR detection in stool samples of HIV-positive and HIV-negative individuals and school children in Vhembe district, Limpopo Province. AB - Microsporidia were initially recognized as pathogens of insects and fish but have recently emerged as an important group of human pathogens, especially in immune compromised individuals, such as those with HIV infection. In this study, we used a PCR-RFLP assay confirmed by quantitative real-time PCR and trichrome staining to determine the prevalence of microsporidian infections among hospital patients and school children in Vhembe region. Enterocytozoon bieneusi was the only microsporidian species detected in these stool samples. It was found in 33 (12.9%) of 255 samples from the hospitals and in 3 (4.5%) of 67 samples from primary school children and was significantly associated (P=0.039) with diarrhea in HIV-positive patients (21.6%) compared to HIV-negative individuals (9%). However, microsporidian infections were not associated with intestinal inflammation as indicated by the lactoferrin test. These results suggest that microsporidia might be a cause of secretory diarrhea in HIV-positive patients. To our knowledge, this is the first report of E. bieneusi in the Vhembe region of South Africa. Further investigations are needed in order to clarify the pathogenesis of E. bieneusi in HIV-positive patients. PMID- 17412379 TI - Studies on the regioselectivity of the Baeyer-Villiger reaction of 3-keto steroids: conformational effects determine the migration aptitude. AB - A detailed study of the Baeyer-Villiger reaction of 3-ketosteroids has been performed by using m-chloroperoxybenzoic and trifluoroperoxyacetic acids as oxidants. The process was fully regiospecific for 3-keto-5alpha-steroids with the employ of both peracids, and only partially regioselective for 3-keto-5beta steroids by using trifluoroperoxyacetic acid. Interestingly, the reaction resulted completely unselective for 3-keto-5beta-steroids by using m chloroperoxybenzoic acid. Theoretical studies were performed to explain the regiochemistry of this process, which is suggested to be controlled by conformational effects in the transition state of the Criegee rearrangement. PMID- 17412380 TI - Golgi complex disassembly caused by light-activated calphostin C involves MAPK and PKA. AB - We examined the participation of MAPK and PKA in the Golgi complex disassembly caused by light-activated Calphostin C in HT-29 cells. When these cells were incubated with Calphostin C, fragmentation and dispersal of the Golgi complex was observed as assessed by immunofluorescence microscopy. Electron microscopy analysis showed that clusters of vesicles and large tubule-vesicular membrane structures, resembling the Golgi remnants present in mitotic cells, substituted the Golgi stacks. In addition, Calphostin C treatment caused inhibition of the endocytic route. We confirmed that the Golgi disassembly was not due to PKC inhibition, and suggested, based on the use of specific inhibitors, that other kinases are involved. It was shown that pretreatment with PD98059 and H-89, both inhibitors of MAPK and PKA, respectively, prior to incubation with Calphostin C, caused blockade of the Golgi disassembly, as well as the inhibition of the endocytic pathway caused by this drug. This finding supports the existence of a novel mechanism by which MAPK and PKA may regulate the Golgi breakdown caused by Calphostin C in HT-29 cells. PMID- 17412381 TI - Ultrastructure of spermiogenesis and spermatozoa of Gymnotus cf. anguillaris and Brachyhypopomus cf. pinnicaudatus (Teleostei: Gymnotiformes). AB - The ultrastructure of spermiogenic stages and spermatozoa of representatives of two gymnotiform families, Gymnotus cf. anguillaris (Gymnotidae) and Brachyhypopomus cf. pinnicaudatus (Hypopomidae) were studied. Spermiogenesis of both species is characterized by lateral development of the flagellum and formation of a nuclear fossa. Some differences were found between these species, such as whether (B. cf. pinnicaudatus) or not (G. cf. anguillaris) nuclear rotation occurs, permanence of the cytoplasmic channel, and type and localization of the nuclear fossa. In the G. cf. anguillaris spermatozoon the nucleus is spherical with highly condensed chromatin. The nuclear fossa is shallow and lateral and is associated with the centriolar complex through stabilizing fibrils. The midpiece is short, with many vesicles, a cytoplasmic channel, and elongate mitochondria. In the B. cf. pinnicaudatus spermatozoon the ovoid nucleus is elongated lateral and posterior to the centriolar complex, and has highly condensed chromatin. The eccentric nuclear fossa is of the moderate type, and contains the entire centriolar complex. The midpiece is long, with numerous vesicles, elongate mitochondria, and no cytoplasmic channel. In both species the flagella are laterally disposed in relation to the nucleus and comprise of the classical 9+2 axoneme. Most of the characteristics found in the spermatozoa of these two species of Gymnotiformes are shared with species of Characiformes, whereas only a few are also found in Siluriformes. This suggests that Gymnotiformes and Characiformes may be more closely related than previously proposed. PMID- 17412382 TI - Biochemical and ultrastructural changes of the liver and kidney of the phytoplanktivorous silver carp feeding naturally on toxic Microcystis blooms in Taihu Lake, China. AB - Many experimental studies have documented the impact of microcystins (MC) on fish based on either intraperitoneal injection, or oral gavaging via the diet, but few experiments were conducted by MC exposure through natural food uptake in lakes. In this study, the phytoplanktivorous silver carp were stocked in a large pen set in Meiliang Bay of Taihu Lake where toxic Microcystis blooms occurred in the warm seasons. Fish samples were collected monthly and MC concentrations in liver and kidney of the fish were determined by LC-MS. The maximum MC concentrations in liver and kidney were present in July when damages in ultrastructures of the liver and kidney were revealed by electron microscope. In comparison with previous studies on common carp, silver carp showed less damage and presence of lysosome proliferation in liver and kidney. Silver carp might eliminate or lessen cell damage caused by MC through lysosome activation. Recovery in the ultrastructures of liver and kidney after Microcystis blooms was companied with a significant decrease or even disappearance of MC. Catalase and glutathione S transferase in liver and kidney of silver carp during Microcystis blooms were significantly higher than before and after Microcystis blooms. The high glutathione pool in liver and kidney of silver carp suggests their high resistance to MC exposure. The efficient antioxidant defence may be an important mechanism of phytoplanktivorous fish like silver carp to counteract toxic Microcystis blooms. PMID- 17412383 TI - Molecular isoforms of cobra venom factor-like proteins in the venom of Austrelaps superbus. AB - Cobra venom factor (CVF) is characteristic of the elapid cobras and has not been reported from venoms of any other families of snakes. During our search for novel proteins, we isolated a polypeptide from the venom of the snake Austrelaps superbus (Lowland Copperhead) that showed structural similarity to C-terminal segment of the alpha-chain of CVF and hence named as AVFalphac (AVF-A. superbus venom factor). cDNA sequence of AVFalphac and its precursor indicated the presence of two isoforms of CVF-like proteins in A. superbus venom gland. This is the first report of molecular isoforms of CVF-like proteins in the venom of an Australian elapid snake. We have determined the complete cDNA sequence of both the isoforms (AVF-1 and AVF-2). They differ in their potential glycosylation sites and the characteristic thioester bond sequence. They display the overall domain structure of CVF and complement C3 proteins. By real-time quantitative analysis, we show that there is a 140-fold difference in the mRNA expression levels of the two isoforms in the venom gland of A. superbus. We also show the presence of AVF-1 and its variant (not AVF-2) in A. superbus venom by partial purification, dot blots, Western blots and peptide mapping using mass spectrometry. Partially purified proteins activate human Factor B in the presence of Factor D and Mg(2+), and deplete the complement activity in human and guinea pig serum. The bimolecular complex (AVFBb) formed activates complement C3 but not complement C5. Thus, AVF proteins may serve as potential candidates for therapeutic complement depletion without side effects. Thus, the discovery of CVF like proteins in the venom of this Australian elapid snake provides an alternative source of research tools, and contributes to our understanding of the structure-function relationships and evolution of new members of CVF-like proteins. PMID- 17412384 TI - Genome of the Acidianus bottle-shaped virus and insights into the replication and packaging mechanisms. AB - The Acidianus bottle-shaped virus, ABV, infects strains of the hyperthermophilic archaeal genus Acidianus and is morphologically distinct from all other known viruses. Its genome consists of linear double-stranded DNA, containing 23,814 bp with a G+C content of 35%, and it exhibits a 590-bp inverted terminal repeat. Of the 57 predicted ORFs, only three produced significant matches in public sequence databases with genes encoding a glycosyltransferase, a thymidylate kinase and a protein-primed DNA polymerase. Moreover, only one homologous gene is shared with other sequenced crenarchaeal viruses. The results confirm the unique nature of the ABV virus, and support its assignment to the newly proposed viral family the Ampullaviridae. Exceptionally, one region at the end of the linear genome of ABV is similar in both gene content and organization to corresponding regions in the genomes of the bacteriophage varphi29 and the human adenovirus. The region contains the genes for a putative protein-primed DNA polymerase, and a small putative RNA with a predicted secondary structure closely similar to that of the prohead RNA of bacteriophage varphi29. The apparent similarities in the putative mechanisms of DNA replication and packaging of ABV to those of bacterial and eukaryal viruses are most consistent with the concept of a primordial gene pool as a source of viral genes. PMID- 17412385 TI - Identification and functional analysis of sequence variants in the long control region and the E2 open reading frame of bovine papillomavirus type 1 isolated from equine sarcoids. AB - BPV-1 DNA is the predominant viral type detected in equine sarcoids and represents the only reported natural cross species infection of papillomaviruses. In this study, nucleotide variations in the LCR and the E2 regions of equine sarcoid-associated BPV-1 were characterised by sequence analysis. Variants particular to sarcoid BPV-1 were identified in both the LCR and E2 sequence. The functionality of the most common LCR variant was examined in equine and bovine cells. These studies showed that the activity of the variant LCR was higher in equine cells than bovine cells; the activity of the variant LCR in the presence of the E2 variant was similar to the reference/wild-type sequences in equine cells, whereas in bovine cells the variant function was reduced by 50%. These data suggest the viral BPV variants commonly detected in sarcoids have an enhanced function in equine cells compared to their function in bovine cells. PMID- 17412386 TI - Rift Valley fever virus lacking NSm proteins retains high virulence in vivo and may provide a model of human delayed onset neurologic disease. AB - Rift Valley fever virus is a significant human and veterinary pathogen responsible for explosive outbreaks throughout Africa and the Arabian Peninsula. Severe acute disease in humans includes rapid onset hepatic disease and hemorrhagic fever or delayed onset encephalitis. A highly efficient reverse genetics system was developed which allowed generation of recombinant RVF viruses to assess the role of NSm protein in virulence in a rat model in which wild-type RVF virus strain ZH501 (wt-ZH501) results in 100% lethal hepatic disease 2-3 days post infection. While extensive genomic analysis indicates conservation of the NSm coding capability of diverse RVF viruses, and viruses deficient in NSs proteins are completely attenuated in vivo, comparison of wt-ZH501, a reverse genetics generated wt-ZH501 virus (R-ZH501), and R-ZH501 virus lacking the NSm proteins (R-DeltaNSm-ZH501) demonstrated that the NSm proteins were nonessential for in vivo virulence and lethality. Surprisingly, while 44% of R-DeltaNSm-ZH501 infected animals quickly developed lethal hepatic disease similar to wt- and R ZH501, 17% developed delayed onset neurologic disease (lethargy, head tremors, and ataxia) at 13 days post infection. Such infections may provide the basis for study of both RVF acute hepatic disease and delayed onset encephalitic disease in humans. PMID- 17412387 TI - Single independent operator sites are involved in the genetic switch of the Lactobacillus delbrueckii bacteriophage mv4. AB - The lysogeny region of the Lactobacillus delbrueckii bacteriophage mv4 contains two divergently oriented ORFs coding for the Rep (221 aa) and Tec (64 aa) proteins. The transcription of these two genes was analysed by primer extension and Northern blot experiments on lysogenic strains. The location of the transcription initiation sites of rep and tec in the intergenic region allowed the identification of the divergently oriented non overlapping promoters P(rep) and P(tec). Transcriptional fusions analysis showed that Rep negatively regulates the P(tec) promoter and activates its own transcription, and that Tec is a negative regulator of the two promoters. As demonstrated by gel mobility shift assays, the repressor Rep binds to a single specific 17 bp site located between the P(tec) -10 and -35 regions whereas Tec binds to a single specific 40 bp long complex operator site located between the two promoters. The presence of a single specific operator site for each repressor in the intergenic region is an unusual feature. PMID- 17412388 TI - Illusory depth induced by binocular torsional misalignment. AB - This study reports a new depth illusion in which a static flat pattern appears stratified stereoscopically when viewed binocularly with an elevated gaze. Three psychophysical experiments measured perceived relative depth and fixational cyclodisparity (a rotation of one eye's view relative to the other eye's view about the line of sight) when flat patterns drawn with solid or dashed curved lines were fixated at various levels of gaze elevation. Experiments 1 and 2 showed that the patterns drawn with solid lines produced illusory depth only at large gaze elevations (downward and upward). Experiment 3 showed that the magnitude of the illusory depth was correlated with that of fixational cyclodisparity. These results suggest that the illusory depth originates in the binocular torsional misalignment generated by gaze elevation. PMID- 17412389 TI - Reductive immobilization of chromate in water and soil using stabilized iron nanoparticles. AB - Laboratory batch and column experiments were conducted to investigate the feasibility of using a new class of stabilized zero-valent iron (ZVI) nanoparticles for in situ reductive immobilization of Cr(VI) in water and in a sandy loam soil. Batch kinetic tests indicated that 0.08g/L of the ZVI nanoparticles were able to rapidly reduce 34mg/L of Cr(VI) in water at an initial pseudo first-order rate constant of 0.08h(-1). The extent of Cr(VI) reduction was increased from 24% to 90% as the ZVI dosage was increased from 0.04 to 0.12g/L. The leachability of Cr preloaded in a Cr-loaded sandy soil was reduced by nearly 50% when the soil was amended with 0.08g/L of the ZVI nanoparticles in batch tests at a soil-to-solution ratio of 1g: 10mL. Column experiments indicated that the stabilized ZVI nanoparticles are highly deliverable in the soil column. When the soil column was treated with 5.7 bed volumes of 0.06g/L of the nanoparticles at pH 5.60, only 4.9% of the total Cr was eluted compared to 12% for untreated soil under otherwise identical conditions. The ZVI treatment reduced the TCLP leachability of Cr in the soil by 90%, and the California WET (Waste Extraction Test) leachability by 76%. The stabilized ZVI nanoparticles may serve as a highly soil-dispersible and effective agent for in situ reductive immobilization of chromium in soils, groundwater, or industrial wastes. PMID- 17412391 TI - Characterization of residual carbon influencing on de novo synthesis of PCDD/Fs in MSWI fly ash. AB - Using 19 samples of fly ash collected from various MSW incineration facilities, residual carbon was characterized by gasifiable fraction at 450 degrees C (C450), and the correlations with de novo synthesis of PCDD/Fs were experimentally examined. Fly ashes were classified into three groups by the ratio of C450 to total residual carbon. By comparison of CO and CO2 generation patterns with those of reference materials, unburnt carbon of solid waste and activated carbon powder injected into flue gas were identified as a carbon source in fly ash. In the experiment of de novo synthesis of PCDD/Fs, the content of PCDD/F synthesis depended on C450 regardless of the origin of carbon. In addition, the model to predict the content of PCDD/F synthesis, DeltaPCDD/F=0.989.Cu.C450, fitted well with experimental values. PMID- 17412392 TI - Assessment of PCDD/F risk after implementation of emission reduction at a MSWI. AB - Municipal solid waste incinerators (MSWIs) have been shown to be important sources of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). The emission of PCDD/Fs by MSWIs is a controversial subject in human health risk assessment. In this study the effect of a MSWI on a residential area was assessed before and after the installation of an additional treatment system for flue gas. This additional treatment system resulted in a dramatic decrease in PCDD/F concentrations in stack flue gas samples by 99.98%, while the concentrations in air decreased by approximately 50% (36,500 and 0.75 pg I-TEQ m(-3) for air in 1999; 3.5 and 0.38 pg I-TEQ m(-3) in 2002; 1.6 and 0.076 pg I-TEQ m(-3) in 2005 for stack gas and air, respectively). Considering the congener distributions of PCDD/Fs between stack flue gas and air samples, the study area seemed to have been contaminated by other urban sources as well as the MSWI. ISC3 model results support the conclusion that this incinerator became only a minor contributor to the study area after installation of the supplementary systems. This resulted from both proper MSWI operation using modern technology and additional sources of contaminants in this region. Finally, PCDD/F uptake by humans through inhalation of contaminated air was estimated. Assuming that inhalation exposure contributes 10% of total exposure, total exposure was lower than WHO guidelines. These results confirm that proper operation and maintenance of the incinerator led to a reduction in emissions and potential health impacts of PCDD/Fs. PMID- 17412393 TI - Production of coloured glass-ceramics from incinerator ash using thermal plasma technology. AB - Incineration is a major treatment process for municipal solid waste in Taiwan. It is estimated that over 1.5 Mt of incinerator ash are produced annually. This study proposes using thermal plasma technology to treat incinerator ash. Sintered glass-ceramics were produced using quenched vitrified slag with colouring agents added. The experimental results showed that the major crystalline phases developed in the sintered glass-ceramics were gehlenite and wollastonite, but many other secondary phases also appeared depending on the colouring agents added. The physical/mechanical properties, chemical resistance and toxicity characteristic leaching procedure of the coloured glass-ceramics were satisfactory. The glass-ceramic products obtained from incinerator ash treated with thermal plasma technology have great potential for building applications. PMID- 17412394 TI - Mathematical analysis of the whole core injection method accuracy for measuring phenanthrene biodegradation rates in undisturbed marine sediments. AB - Rates of 14C-phenanthrene mineralization in contaminated, undisturbed marine sediments were measured using the whole core injection method to assess microbial natural attenuation activity as a function of sediment depth. Submerged sediments were sampled from Eagle Harbor, a marine superfund site in Puget Sound. Experiments show significant biodegradation activities (0.0012-0.0036 day(-1)) in the sediment horizons from 0 to 10 cm. The purpose and scope of this paper is to evaluate the range of experimental conditions giving valid results; a mathematical simulation described competing contaminant 14C-phenanthrene diffusion and simultaneous biodegradation (Monod kinetics), both retarded by sorption. The effect of aging was examined with two sorption models in presumed pseudo-homogenous sediments having effective properties. The simulation predictions provide quantitative guidelines for the successful use of the whole core injection method. (1) The effective Monod constant KS' in sediment is increased by a large partition coefficient KP between sediment and water and makes the apparent 14C-phenanthrene biodegradation approach first-order kinetics. (2) When KS'>1 mg(-1) l(-1), the measured 14C-phenanthrene biodegradation extent is biased by inadequately distributed injected tracer only when less than 7% of the sediment horizon is initially probed and mixed with injected tracer. (3) A short incubation time (<20 days) is necessary when a mobile indicator, e.g., gaseous 14CO2, is used. For longer incubation times, predictions show that a 14CO2 indicator diffuses to adjacent horizons, thus smearing the depth profile of biodegradation. (4) This method employing a radiolabeled tracer provides accurate biodegradation rates for freshly contaminated sediments, and represents an upper limit to the natural phenanthrene biodegradation extents if the contaminant is aged over 50 days. PMID- 17412395 TI - Hominin homoiology: an assessment of the impact of phenotypic plasticity on phylogenetic analyses of humans and their fossil relatives. AB - Homoiologies are phylogenetically misleading morphological similarities that are due to nongenetic factors. It has been claimed that homoiologies are common in the hominin skull, especially in regions affected by masticatory strain, and that their prevalence is one reason why reconstructing hominin phylogenetic relationships is difficult. To evaluate this "homoiology hypothesis," we performed analyses on a group of extant primates for which a robust molecular phylogeny is available--the hominoids. We compiled a data set from measurements that developmental considerations and experimental evidence suggest differ in their likelihood of exhibiting masticatory-strain-induced phenotypic plasticity. We then used the coefficient of variation and t-tests to evaluate the phenotypic plasticity of the measurements. We predicted that, if the hypothesis is correct, the measurements of skeletal features that do not remodel and therefore are unaffected by phenotypic plasticity should be less variable than the measurements of skeletal features that remodel and are subject to low-to-moderate strains, and that the latter should be less variable than the measurements of skeletal features that remodel and are subject to moderate-to-high strains. Subsequently, we performed phylogenetic analyses on character state data derived from the measurements and compared the resulting phylogenetic hypotheses to the consensus molecular phylogeny for the hominoids. We predicted that, if the hypothesis is correct, agreement between the phylogenies should be best for the non phenotypically-plastic characters, intermediate for the low-to-moderate-strain characters, and worst for the moderate-to-high-strain characters. The results of the coefficient of variation/t-test analyses were consistent with the predictions of the hypothesis to the extent that the moderate-to-high-strain measurements exhibited significantly more variability than the non-phenotypically-plastic and low-to-moderate-strain measurements. In contrast, the results of the phylogenetic analyses were not those predicted. The phylogeny derived from the moderate-to high-strain characters matched the molecular phylogeny better than those obtained using the non-phenotypically-plastic and low-to-moderate-strain characters. Thus, our study supports the suggestion that mechanical loading results in phenotypic plasticity in the hominin skull, but it does not support the notion that homoiologies have a significant negative impact on hominin phylogenetics. PMID- 17412396 TI - Levels and trends of organochlorines and brominated flame retardants in ivory gull eggs from the Canadian Arctic, 1976 to 2004. AB - The ivory gull (Pagophila eburnea) is a circumpolar marine bird which has recently been listed as an endangered species in Canada. To determine whether contaminants may be playing a role in the population decline of this species, ivory gull eggs collected in 1976, 1987 and 2004 from Seymour Island in the Canadian Arctic were analyzed for organochlorines, polychlorinated dibenzo-p dioxins (PCDDs), dibenzofurans (PCDFs) and non-ortho PCBs. This study also provides the first account of polybrominated diphenyl ethers (PBDEs), hexabromocyclododecanes (HBCDs) and polybrominated biphenyls (PBBs) in ivory gulls. The most quantitatively abundant legacy organochlorines found in the ivory gull eggs were p,p'-DDE, SigmaPCB and oxychlordane. Concentrations of the organochlorines analyzed either decreased or showed little change between 1976 and 2004. Concentrations of SigmaPCDD in ivory gull eggs were greater than SigmaPCDF, and the non-ortho PCBs (primarily PCB-126) contributed the largest fraction to the total TEQ value in all years sampled. Concentrations of PCDDs, PCDFs and SigmaTEQ decreased from 1976 to 2004. In contrast, concentrations of the PBDEs steadily increased between 1976 and 2004 driven primarily by increases in BDE-47. Although concentrations of the persistent chlorinated compounds (i.e. organochlorine pesticides, PCBs, PCDDs, PCDFs) reported in this study were below published toxicological threshold values for eggs of wild birds, we cannot rule out the possibility of synergistic/additive, sublethal effects. Very few studies have been carried out to evaluate the exposure-effect relationship of the persistent brominated compounds in avian species. Given the scarcity of information on toxicity threshold levels for PBBs and PBDEs in avian species, coupled with the trend toward increasing concentrations in ivory gulls, continued monitoring and further toxicological studies of these compounds are warranted. PMID- 17412397 TI - Atmospheric deposition of nitrogen at five subtropical forested sites in South China. AB - Elevated concentrations of reactive nitrogen (N) in precipitation have been reported for many cities in China. Due to increased use of fossil fuels and expansion in agriculture, further increases in deposition of ammonia (NHx) and reactive N oxides (NOy) are predicted. Increased deposition of reactive N is likely to affect N dynamics and N runoff in forest ecosystems. Yet, in China little work has been done to quantify the levels of atmospheric N deposition in such systems. Here, we assess the deposition of inorganic N (ammonium, NH4+ and nitrate, NO3-) for five subtropical forest ecosystems in remote and urban areas of South China. Annual volume-weighted concentrations in bulk precipitation range from 0.18 to 1.55 mg NH4+ -N L(-1) and from 0.12 to 0.74 mg NO3- -N L(-1). These values are large and several times greater than those reported for remote sites of the world. The fluxes of total inorganic N (TIN) in wet-only deposition range from 0.8 to 2.3 g N m(-2) yr(-1), with NH4+ -N contributing 54% to 77%. Both the tree canopy and the ground vegetation layer are important in determining the net N flux reaching the forest floor, but the net effect varies from site to site. At TieShanPing (TSP), close to Chongqing city, and at CaiJiaTang (CJT), near Shaoshan (Hunan province), the canopy represents a net source of N, probably due to dry deposition. At the other three sites (LiuChongGuan (LCG), LeiGongShan (LGS), both in Guizhou province, and LiuXiHe (LXH) in Guangdong), a net loss of reactive N from precipitation water occurs in the canopy, probably due to uptake processes. The total annual atmospheric TIN load is estimated to range from at least 0.8 g N m(-2) yr(-1) to 4.0 g N m(-2) yr(-1), with a considerable contribution from dry deposition. Concentrations and fluxes of inorganic N in tree canopy throughfall are greater than those in North America. Also the contribution of NH4+ -N to TIN fluxes in throughfall (40% to 70%) is greater than in North America. Our sites have N deposition levels and NH4+ -N contributions similar to those found in Northwest and central Europe. The TIN loads at TSP and CJT far exceed 2.5 g N m(-2) yr(-1), which is the N deposition load above which NO3- leaching is expected in temperate and boreal forests. PMID- 17412398 TI - Multiple linear regression modeling of disinfection by-products formation in Istanbul drinking water reservoirs. AB - Oxidation of raw water with chlorine results in formation of trihalomethanes (THM) and haloacetic acids (HAA). Factors affecting their concentrations have been found to be organic matter type and concentration, pH, temperature, chlorine dose, contact time and bromide concentration, but the mechanisms of their formation are still under investigation. Within this scope, chlorination experiments have been conducted with water reservoirs from Terkos, Buyukcekmece and Omerli lakes, Istanbul, with different water quality regarding bromide concentration and organic matter content. The factors studied were pH, contact time, chlorine dose, and specific ultraviolet absorbance (SUVA). The determination of disinfection by-products (DBP) was carried out by gas chromatography techniques. Statistical analysis of the results was focused on the development of multiple regression models for predicting the concentrations of total THM and total HAA based on the use of pH, contact time, chlorine dose, and SUVA. The developed models provided satisfactory estimations of the concentrations of the DBP and the model regression coefficients of THM and HAA are 0.88 and 0.61, respectively. Further, the Durbin-Watson values confirm the reliability of the two models. The results indicate that under these experimental conditions which indicate the variations of pH, chlorine dosages, contact time, and SUVA values, the formation of THM and HAA in water can be described by the multiple linear regression technique. PMID- 17412399 TI - Dietary exposure to organophosphorus pesticides for young children in Tokyo and neighboring area. AB - Dietary ingestion is a significant pathway of human exposure to organophosphorus (OP) pesticides. This study aims to determine the occurrence of OP pesticide in the diet of young children in Japan and estimate their exposure. The study was conducted by analyzing duplicate diet samples collected from 33 children aged 3 to 6 years old in Tokyo and the nearby area in Japan. Seven OP pesticides that are mainly used for agriculture in Japan (fenitrothion, trichlorfon, dichlorvos, diazinon, chlorpyrifos, malathion, and EPN) were targeted for analysis. As a result of the study, fenitrothion was frequently found in the week's diet (21%) of children, followed by diazinon (14%), chlorpyrifos (11%), and malathion (4%); the daily intake of pesticides was also estimated to range from < 110 to 180 ng/day, < 70 to 800 ng/day, < 70 to 100 ng/day, and < or = 110 ng/day, respectively. Daily intake of diazinon per kg body weight was relatively higher than other target pesticides and the maximum value was 50 times below the acceptable daily intake set by Japan Ministry of Health and Labor Welfare. Occurrence of malathion in the diet of children in the present study was lower than that reported in 1980s, supposed due to the decreasing level of malathion in wheat and wheat products. Frequent detection of fenitrothion in diet is a significant trend observed in Japan, and is considered due to the high and widespread usage of the pesticide inside of the country. PMID- 17412400 TI - Are responses to treatment virus-specific in wheezing children? PMID- 17412401 TI - Advances in allergic skin disease, anaphylaxis, and hypersensitivity reactions to foods, drugs, and insects. AB - This review highlights some of the research advances in anaphylaxis and hypersensitivity reactions to foods, drugs, and insects and in allergic skin disease that were reported primarily in the Journal in 2006. Advances in diagnosis include identification of food proteins to which IgE binding is associated with severe reactions; elucidation of diagnostic relationships of skin prick test wheal size with outcomes of egg, tree nut, and sesame allergy; evaluation of the diagnostic utility of atopy patch testing for food; and the observation that yellow jacket sting outcomes are influenced by species. Mechanistic observations include the following: heating of birch pollen-related foods disrupts IgE binding but not T-cell epitopes; a simple imbalance of T(H)1/T(H)2 response does not explain variations in clinical expression of peanut allergy; and elucidation of the role of dendritic cells in drug hypersensitivity. With regard to treatment, a rapidly disintegrating epinephrine tablet showed promise for sublingual treatment of anaphylaxis, RNA interference techniques showed promise in creating lower-allergenic foods, and anti-IL-5 showed promise for treatment of eosinophilic esophagitis. Progress in our understanding of the immunology and the etiology of skin barrier dysfunction in atopic dermatitis has also been made. These observations will likely contribute toward optimizing management of these common allergic disorders. PMID- 17412402 TI - Quantitative and functional impairment of pulmonary CD4+CD25hi regulatory T cells in pediatric asthma. AB - BACKGROUND: Asthma is characterized by a T(H)2 immune response. CD4(+)CD25(hi) regulatory T cells (Tregs) have been proposed to prevent allergic diseases through suppression of T(H)2 responses. OBJECTIVE: We sought to investigate the role of CD4(+)CD25(hi) T cells in children with asthma. METHODS: CD4(+)CD25(hi) Tregs and forkhead/winged-helix transcription factor FOXP3 mRNA levels were quantified in peripheral blood and bronchoalveolar lavage fluid (BALF) of 18 children with asthma, 10 children with chronic cough, and 13 control subjects without lung diseases. CD4(+)CD25(hi) T cells were isolated from peripheral blood and BALF of asthmatic patients and control subjects, and their capacity to suppress proliferation and cytokine/chemokine production of autologous responder T cells was analyzed. RESULTS: CD4(+)CD25(hi) T cells were decreased in BALF of asthmatic children compared with values in children with cough or control subjects. In children with asthma, inhaled corticosteroid treatment was associated with increased percentages of CD4(+)CD25(hi) T cells in peripheral blood and BALF. Isolated BALF and peripheral blood CD4(+)CD25(hi) T cells from nonasthmatic subjects suppressed proliferation and cytokine/chemokine production by CD4(+)CD25(-) responder T cells. BALF CD4(+)CD25(hi) T cells from asthmatic subjects failed to suppress proliferation and production of T(H)2-associated cytokines and chemokines by CD4(+)CD25(-) responder T cells, which was restored after use of inhaled corticosteroids. CONCLUSION: These findings provide the first evidence that pulmonary CD4(+)CD25(hi) Tregs are impaired in pediatric asthma. CLINICAL IMPLICATIONS: Pulmonary Tregs might represent a therapeutic target in pediatric asthma. PMID- 17412403 TI - Prenatal versus postnatal sensitization to environmental allergens in a high-risk birth cohort. AB - BACKGROUND: The timing of allergen sensitization is controversial, with conflicting evidence suggesting transplacental priming versus exclusively postnatal priming. Resolution of this question is important in relation to rational design of allergy prevention strategies, particularly the issue of allergen avoidance during pregnancy. OBJECTIVE: To elucidate the kinetics of sensitization in high-risk children during their first 2 years of life. METHODS: We prospectively studied house dust mite (HDM)-specific IgE and IgG(4) antibody production and associated T-cell immunity in a cohort of 200 high-risk infants. Parallel antibody studies tracked responses against a broader panel of inhalant and dietary allergens including peanut. RESULTS: HDM-induced T(H)2 responses in PBMC from 6 months onward, particularly IL-4 and IL-5, correlated increasingly strongly with sensitization outcomes at 2 years, and a contrasting negative relationship was observed with IFN-gamma response capacity. HDM-induced T-cell responses in cord blood, although common, were unrelated to subsequent sensitization. Transient HDM-IgE (and IgG(4)) production frequently peaked at 6 or 12 months before returning to baseline, which suggests the onset of protective tolerance. This finding contrasted with progressively increasing HDM-IgE titers in children sensitized by 2 years of age. Comparably contrasting patterns were observed in peanut-specific responses in sensitized versus nonsensitized children. CONCLUSION: Priming of T(H)2 responses associated with persistent HDM IgE production occurs entirely postnatally, as HDM reactivity in cord blood seems nonspecific and is unrelated to subsequent development of allergen-specific T(H)2 memory or IgE. CLINICAL IMPLICATIONS: These findings question the scientific basis for existing recommendations for allergen avoidance by high-risk women during pregnancy. PMID- 17412404 TI - Cross-reactivity patterns of T cells specific for iodinated contrast media. AB - BACKGROUND: Approximately 3% of patients exposed to iodinated contrast media develop delayed hypersensitivity reactions. OBJECTIVE: We wanted to better understand the molecular basis of contrast media cross-reactivity. METHODS: Cross reactivity was assessed by skin testing and measurement of T-cell activation (CD69 upregulation) and proliferation ((3)H-thymidine uptake, 5,6 carboxyfluorescein diacetate succinimidyl ester staining) of PBMCs, T-cell lines, and T-cell clones of 2 patients with delayed hypersensitivity reactions to iohexol and iomeprol, respectively. Thirteen different contrast media and potassium iodide were compared. RESULTS: Skin testing and analyses of PBMCs, T cell lines, and clones showed broad cross-reactivity in both patients. Broad as well as more restricted cross-reactivity patterns were found in iohexol-specific and iomeprol-specific CD4(+) T-cell clones, whereas 1 iomeprol-specific CD8(+) T cell clone showed no cross-reactivity at all. The reactivity to equimolar concentrations of iohexol and its dimer iodixanol was very similar, suggesting that the dimer was not more stimulatory than its monomer. Consistently low reactivity to iobitridol was found in both patients, but never to iodide. A frequency analysis of contrast medium-specific peripheral T cells gave values between 0.6 % (iomeprol) and 0.05 % (iobitridol). CONCLUSION: Clinically observed cross-reactivity between different contrast media is a result of the presence of contrast media-specific T cells, some of which show a broad cross-reactivity pattern. Iodide ions, known to be present at low concentration in contrast media solutions, do not seem to be the causative moiety. CLINICAL IMPLICATIONS: Detailed in vitro analysis might help identify noncross-reactive contrast media. PMID- 17412405 TI - Connective tissue growth factor expression is regulated by histamine in lung fibroblasts: potential role of histamine in airway remodeling. AB - BACKGROUND: In the inflamed lung of allergic asthma, an aberrant injury-repair response is accompanied by structural changes in the airway, known as airway remodeling. TGF-beta and its downstream mediator connective tissue growth factor (CTGF) are playing a key role in these processes, resulting in irreversible airway remodelling. OBJECTIVE: As histamine is a key mediator of allergic reactions, we investigated whether histamine is involved in airway remodeling. METHODS: The effect of histamine and TGF-beta1 on proliferation of lung fibroblast cells IMR-90 was studied by [(3)H]-thymidine proliferation assay. The regulation of CTGF by histamine and TGF-beta1 in lung fibroblasts was analyzed by RT-PCR, real-time PCR, Western blot analysis, and promoter analysis and characterized by specific histamine-receptor antagonists. RESULTS: Histamine and TGF-beta1 enhanced proliferation of lung fibroblast cells IMR-90. Both induced CTGF mRNA and protein expression with different time kinetics. Whereas TGF-beta1 induced maximal CTGF expression after 12 hours (347% +/- 23%), histamine-induced maximal CTGF expression was lower and delayed (maximum expression of 204% +/- 11% after 48 hours). Histamine and TGF-beta1 stimulated the CTGF promoter and the TGF beta-response element in the CTGF promoter. The histamine-induced CTGF expression was mediated through the histamine receptor (HR1) and could be completely abolished by TNF-alpha. CONCLUSIONS: These findings demonstrate that histamine plays a potential role in the induction of airway remodeling mediated by the induction of lung fibroblasts proliferation and CTGF expression. CLINICAL IMPLICATIONS: This mechanism could be important for prophylactic strategies aiming at airway remodeling and could be a new indication for antihistamine treatment. PMID- 17412406 TI - Images in allergy and immunology: neutrophils in asthma. PMID- 17412407 TI - Limited IgE cross-reactivity between Dermatophagoides pteronyssinus and Glycyphagus domesticus in patients naturally exposed to both mite species. AB - BACKGROUND: Contradictory results have been found when analyzing the IgE cross reactivity among pyroglyphid mites and storage mites, as well as the role of these as true sensitizing agents. OBJECTIVE: We sought to study the prevalence of sensitization to Glycyphagus domesticus in patients naturally exposed to this mite together with the more ubiquitous Dermatophagoides pteronyssinus and the IgE cross-reactivity between them. METHODS: Mite species present in house dust samples of consecutive patients with mite allergy from Huelva (southwest Spain) were determined by means of light microscopy. Clinical sensitization was assessed by using skin prick and conjunctival provocation tests. Cross-reactivity at the IgE level was studied by using serum IgE determination and inhibition of RAST, IgE immunoblotting, and mite group 2 immunoassays. RESULTS: After D pteronyssinus, which is present in about 95% of house dust samples, G domesticus was the most important mite and present in about 50% of the samples. Tyrophagus putrescentiae and Lepidoglyphus destructor were detected in third and fourth place, respectively. About half of the patients with G domesticus at home were sensitized to this mite. A low IgE cross-reactivity was observed between D pteronyssinus and G domesticus, but an important IgE cross-reactivity was detected among glycyphagid mites at the level of group 2 allergens. CONCLUSION: Glycyphagid mites can act as primary sensitizing agents independently of pyroglyphid mites in a subset of patients naturally exposed to them. CLINICAL IMPLICATIONS: The inclusion of glycyphagid mite extracts in the diagnostic battery in areas with adequate mite growing conditions is important to ensure proper diagnosis. PMID- 17412408 TI - Identification of the FANCI protein, a monoubiquitinated FANCD2 paralog required for DNA repair. AB - Fanconi anemia (FA) is a developmental and cancer-predisposition syndrome caused by mutations in genes controlling DNA interstrand crosslink repair. Several FA proteins form a ubiquitin ligase that controls monoubiquitination of the FANCD2 protein in an ATR-dependent manner. Here we describe the FA protein FANCI, identified as an ATM/ATR kinase substrate required for resistance to mitomycin C. FANCI shares sequence similarity with FANCD2, likely evolving from a common ancestral gene. The FANCI protein associates with FANCD2 and, together, as the FANCI-FANCD2 (ID) complex, localize to chromatin in response to DNA damage. Like FANCD2, FANCI is monoubiquitinated and unexpectedly, ubiquitination of each protein is important for the maintenance of ubiquitin on the other, indicating the existence of a dual ubiquitin-locking mechanism required for ID complex function. Mutation in FANCI is responsible for loss of a functional FA pathway in a patient with Fanconi anemia complementation group I. PMID- 17412409 TI - Climate factors affecting conception rate of high producing dairy cows in northeastern Spain. AB - Summer heat stress is a main factor related to low conception rate in high producing dairy herds in warm areas worldwide. We assessed the impact of several climate variables on conception rate in high producing dairy cows in northeastern Spain by examining 10,964 inseminations. The temperature-humidity index (THI) was compared with maximum temperature in terms of its efficiency at predicting conception rate. The following data were recorded for each animal: herd, lactation number, insemination number, insemination date, inseminating bull, and AI technician along with climate variables such as mean and maximum temperatures, rainfall, mean and maximum THI for individual time points Days 7 to 1 before insemination, the day of insemination and 1, 2 and 3 days after insemination. Averages were also established for the following periods: from 7 days before insemination to the insemination day, from 3 days before insemination to the insemination day and from the insemination day to 3 days postinsemination. Based on the odds ratios, the likelihood of conception rate increased significantly by factors of 1.48, 1.47, 1.5, and 1.1 for the respective maximum THI classes <70, 71-75, 76-80, and 81-85 only on Day 3 before AI, while on the insemination day, it increased by factors of 1.73, 1.53, 1.11, and 1.3 for the respective maximum THI classes <70, 71-75, 76-80, and 81-85. In a subsequent logistic regression excluding mean and maximum THI, the effectiveness of temperature at predicting conception rate was evaluated. Although high, the fit of the second logistic regression model was slightly lower than that of the full model (P=0.88 versus P=0.98, respectively) and the information provided by the THI model. The likelihood of conception rate increased significantly by factors of 1.5, 1.2, 1.0, 1.0 for the respective maximum temperature classes <20, 21-25, 26-30, 31-35 degrees C on Day 1 after AI. The choice of the THI or temperature to monitor the farm environment would have to depend on the particular farm and situation. In our study conditions, the use of maximum temperature alone gives a new point of view regarding the information provided by the THI variables. PMID- 17412410 TI - Duration of auditory sensory memory in parents of children with SLI: a mismatch negativity study. AB - In a previous behavioral study, we showed that parents of children with SLI had a subclinical deficit in phonological short-term memory. Here, we tested the hypothesis that they also have a deficit in nonverbal auditory sensory memory. We measured auditory sensory memory using a paradigm involving an electrophysiological component called the mismatch negativity (MMN). The MMN is a measure of the brain's ability to detect a difference between a frequent standard stimulus (1000 Hz tone) and a rare deviant one (1200 Hz tone). Memory effects were assessed by varying the inter-stimulus interval (ISI) between the standard and deviant. We predicted that parents of children with SLI would have a smaller MMN than parents of typically developing children at a long ISI (3000 ms), but not at a short one (800 ms). This was broadly confirmed. However, individual differences in MMN amplitude did not correlate with measures of phonological short-term memory. Attenuation of MMN amplitude at the longer ISI thus did not provide unambiguous support for the hypothesis of a reduced auditory sensory memory in parents of affected children. We conclude by reviewing possible explanations for the observed group effects. PMID- 17412411 TI - Molecular aspects of preeclampsia. AB - Various hypotheses as to the origin of preeclampsia have been explored over time. Diseases of pregnancy are difficult to study for several reasons. One limitation is due to the fact that preeclampsia and associated diseases clinically present in the second and third trimenon, but seem to originate early in pregnancy. Comparisons with animal models are difficult due to the unique human nature of the disease. The creation of new methods including proteomics, genomics, lipidomics, metabolomics or mRNA microarray techniques supplement the traditional type of research access to approach mother and fetus. The clinical course will be discussed and pregnancy-related processes, which are thought to contribute to the disease. This includes implantation of the placenta/fetus, the adaptation of the endothelial activity to the pregnancy with respect to relaxin, matrix metalloproteinases and endothelin, nitric oxide, angiogenetic factors and TGF-b in normal and preeclamptic pregnancies. Furthermore, oxidative stress, genetics and hypothesis-generating molecular approaches are considered. PMID- 17412412 TI - Differential scanning calorimetry (DSC) and temperature-modulated DSC study of three mouthguard materials. AB - OBJECTIVES: Employ differential scanning calorimetry (DSC) and temperature modulated DSC (TMDSC) to investigate thermal transformations in three mouthguard materials and provide insight into their previously investigated energy absorption. METHODS: Samples (13-21mg) were obtained from (a) conventional ethylene vinyl acetate (EVA), (b) Pro-form, another EVA polymer, and (c) PolyShok, an EVA polymer containing polyurethane. Conventional DSC (n=5) was first performed from -80 to 150 degrees C at a heating rate of 10 degrees C/min to determine the temperature range for structural transformations. Subsequently, TMDSC (n=5) was performed from -20 to 150 degrees C at a heating rate of 1 degrees C/min. Onset and peak temperatures were compared using ANOVA and the Tukey-Kramer HSD test. Other samples were coated with a gold-palladium film and examined with an SEM. RESULTS: DSC and TMDSC curves were similar for both conventional EVA and Pro-form, showing two endothermic peaks suggestive of melting processes, with crystallization after the higher-temperature peak. Evidence for crystallization and the second endothermic peak were much less prominent for PolyShok, which had no peaks associated with the polyurethane constituent. The onset of the lower-temperature endothermic transformation is near body temperature. No glass transitions were observed in the materials. SEM examination revealed different surface morphology and possible cushioning effect for PolyShok, compared to Pro-form and EVA. SIGNIFICANCE: The difference in thermal behavior for PolyShok is tentatively attributed to disruption of EVA crystal formation, which may contribute to its superior impact resistance. The lower-temperature endothermic peak suggests that impact testing of these materials should be performed at 37 degrees C. PMID- 17412413 TI - An assessment of the impact of copper mine tailings disposal on meiofaunal assemblages using microcosm bioassays. AB - Microcosms were used to assess the impact of copper mine tailings disposal on the littoral meiofaunal assemblages of the Atacama region of northern Chile. The specific purpose was to establish a cause and effect relationship between the elevated copper concentrations and altered meiofaunal assemblages observed at the study sites. Meiofaunal assemblages were exposed to a series of copper concentrations to assess general toxicity, both densities and taxa diversities decreased with increasing copper. Natural coarse sediments were mixed with a tailings substitute to assess the physical impact of the tailings dumping on meiofaunal assemblages. Meiofaunal assemblage densities increased with increasing amounts of tailings substitute, entirely due to an increase in surface utilising foraminiferans. However, taxa diversities decreased as the interstitial spaces became blocked. Finally, the microcosms were used to conduct bioassays of sediments and seawaters from the impacted sites. The sediments from the impacted sites proved to be toxic resulting in reduced meiofaunal densities and taxa diversities. Seawater samples did not prove to be significantly toxic. The use of microcosms has allowed the effects of the physical and chemical components of tailings to be assessed individually, which was not possible in the field. Additionally, it allowed a cause and effect relationship to be established between elevated concentrations of porewater copper observed in the field and the reduced densities and taxa diversities of the meiofaunal assemblages observed at the same sites. PMID- 17412414 TI - Osteochondral repair using the combination of fibroblast growth factor and amorphous calcium phosphate/poly(L-lactic acid) hybrid materials. AB - A novel amorphous calcium phosphate (ACP)/poly(L-lactic acid) (PLLA) material, which can experience morphological variations in the microstructure is supposed to be a suitable candidate as scaffold for cartilage tissue-engineering. The purpose of this study was to evaluate the efficacy of this scaffold combined with basic fibroblast growth factor (bFGF) to repair articular cartilage defects in a rabbit model. Forty-two osteochondral defects created in the femoral condyles were (a) left untreated, (b) treated by PLLA combined with bFGF, or (c) ACP/PLLA loaded with bFGF. The treatment of PLLA incorporated with bFGF improved defect filling compared with that left untreated, while the regenerated tissue was mainly fibrocartilage and showed little bone formation with only a small amount of collagen type II (Col II) and no aggrecan gene message measured. When implanted with ACP/PLLA and bFGF, most of the defects were filled with a well established layer of cartilage tissue with abundance of cartilaginous extracellular matrix accumulation observed. Positive immunohistochemical staining of Col II was observed. High levels of Col II and aggrecan message were also detected by RT-PCR. These results indicate the feasibility of using the combination of ACP/PLLA with bFGF for cartilage repair. PMID- 17412415 TI - The performance of poly-epsilon-caprolactone scaffolds in a rabbit femur model with and without autologous stromal cells and BMP4. AB - The ability of a cellular construct to guide and promote tissue repair strongly relies on three components, namely, cell, scaffold and growth factors. We aimed to investigate the osteopromotive properties of cellular constructs composed of poly-epsilon-caprolactone (PCL) and rabbit bone marrow stromal cells (BMSCs), or BMSCs engineered to express bone morphogenetic protein 4 (BMP4). Highly porous biodegradable PCL scaffolds were obtained via phase inversion/salt leaching technique. BMSCs and transfected BMSCs were seeded within the scaffolds by using an alternate flow perfusion system and implanted into non-critical size defects in New Zealand rabbit femurs. In vivo biocompatibility, osteogenic and angiogenic effects induced by the presence of scaffolds were assessed by histology and histomorphometry of the femurs, retrieved 4 and 8 weeks after surgery. PCL without cells showed scarce bone formation at the scaffold-bone interface (29% bone/implant contact and 62% fibrous tissue/implant contact) and scarce PCL resorption (16%). Conversely, PCL seeded with autologous BMSCs stimulated new tissue formation into the macropores of the implant (20%) and neo-tissue vascularization. Finally, the BMP4-expressing BMSCs strongly favoured osteoinductivity of cellular constructs, as demonstrated by a more extensive bone/scaffold contact. PMID- 17412416 TI - In vitro bioactivity and gene expression by cells cultured on titanium dioxide doped phosphate-based glasses. AB - In our previous study, glasses with 50 P(2)O(5)-(20-15) Na(2)O-30 CaO-(0-5 mol%) TiO(2) have been prepared by the conventional melt-quenching process. MG63 cell proliferation, gene expression, in vivo biocompatibility, and bioactivity of these glasses is the concern of this study. The results showed that addition of TiO(2) in small amounts up to 5 mol% enhanced the biocompatibility of these glasses. The cell metabolic activity was conspicuous, on 3 and 5 mol% TiO(2) compositions in particular, with no significant difference from Thermanox control over a period of 21 days. The findings from the gene expression study showed that, at day 1 and on 5 mol% TiO(2) glass, core binding protein factor alpha 1 (Cbfa1) and alkaline phosphatase (ALP) showed significantly lower transcription level; however, collagen type I alpha subunit I (COLIAI) and Osteonectin (Sparc) showed no significant differences compared to the control. At day 7, all these genes transcription levels were not significantly different form the control, but at day 14, they were significantly higher than the control. Moreover, there were no significant differences detected in these genes on both 3 and 5 mol% TiO(2) glasses up to 7 days. At day 14; however, 5 mol% TiO(2) glasses showed significantly higher level than 3 mol% TiO(2) composition. This was also correlated by the presence of new bone tissue at the bone-particles interface for 5 mol% TiO(2) composition after 5 weeks of implantation in rat calvarium. Regardless of this favourable cell response and gene up-regulation, these glasses showed no evidence of apatite layer formation after 14 days incubation in SBF. PMID- 17412417 TI - Systemic ablation of RyR3 alters Ca2+ spark signaling in adult skeletal muscle. AB - Ca2+ sparks are localized intracellular Ca2+ release events from the sarcoplasmic reticulum in muscle cells that result from synchronized opening of ryanodine receptors (RyR). In mammalian skeletal muscle, RyR1 is the predominant isoform present in adult skeletal fibers, while some RyR3 is expressed during development. Functional studies have revealed a differential role for RyR1 and RyR3 in the overall Ca2+ signaling in skeletal muscle, but the contribution of these two isoforms to Ca2+ sparks in adult mammalian skeletal muscle has not been fully examined. When enzyme-disassociated, individual adult skeletal muscle fibers are exposed to an osmotic shock, the resting fiber converts from a quiescent to a highly active Ca2+ release state where Ca2+ sparks appear proximal to the sarcolemmal membrane. These osmotic shock-induced Ca2+ sparks occur in ryr3(-/-) muscle with a spatial distribution similar to that seen in wild type muscle. Kinetic analysis reveals that systemic ablation of RyR3 results in significant changes to the initiation, duration and amplitude of individual Ca2+ sparks in muscle fibers. These changes may reflect the adaptation of the muscle Ca2+ signaling or contractile machinery due to the loss of RyR3 expression in distal tissues, as biochemical assays identify significant changes in expression of myosin heavy chain protein in ryr3(-/-) muscle. PMID- 17412418 TI - Presence of peripheral blasts in refractory anemia and refractory cytopenia with multilineage dysplasia predicts an unfavourable outcome. AB - The World Health Organization (WHO) assigns myelodysplastic syndrome (MDS) to RA/RCMD/RARS/RSCM/5q- syndrome, if medullary blasts are <5% and peripheral blast (PB) count < or =1%. In 1103 patients with these diagnoses, we analysed survival and risk of AML evolution depending on the presence of PB. Median survival in the group with 1% PB (n=74) was significantly lower as compared to those without PB (20 versus 47 months, p<0.00005). Cumulative risk of AML was significantly higher in patients showing PB (p<0.00005). Median survival of patients with PB was not different from that of RAEB I. We therefore propose to consider patients with PB, regardless of medullary blast, as RAEB I. PMID- 17412419 TI - Y253H mutation appearing in a micro-BCR-ABL (e19a2) CML. PMID- 17412420 TI - Differential correlates of multi-type maltreatment among urban youth. AB - OBJECTIVE: The aim of this study was to examine the differential effects of multi types of maltreatment in an adolescent sample. Different combinations of maltreatment (emotional, sexual, physical, neglect) were examined in relation to both negative affect and externalizing symptoms in male and female youth. METHOD: One thousand four hundred fifty-two middle and high school youth were recruited from urban schools and a mandated early warning truancy program. Youth completed an anonymous survey that included measures of child maltreatment, depression, suicide proneness, hopelessness, delinquency, hostility, substance use, and promiscuity. Respondents were categorized into groups of different combinations of maltreatment by their reports of sexual abuse, physical abuse, neglect (emotional and physical), and emotional abuse. RESULTS: Nearly two-thirds of boys and girls reported some form of maltreatment, and multi-type maltreatment was common (e.g., 13% reported experiencing both physical and sexual abuse and neglect). Individuals with maltreatment histories were more depressed (F=52.78, p<.0001), suicide prone (F=24.29, p<.001), and hopeless (F=32.07, p<.0001) than non-abused individuals. Maltreated adolescents were also more hostile (F=35.03, p<.0001), and they engaged in more delinquent behavior (F=26.76, p<.0001), promiscuity (F=8.54, p<.0001), and drug and alcohol use (F=9.61, p<.0001). Individuals experiencing multi-type maltreatment were the most symptomatic, particularly youth with histories of physical abuse, sexual abuse, and neglect. In general, gender differences in effects were not observed. CONCLUSIONS: The results highlight the importance of studying combined types of maltreatment, as well as understanding the particularly deleterious effects of neglect and emotional abuse. The results are generally consistent with an additive model of maltreatment effects. PMID- 17412421 TI - The influence of career experience in defining head injuries: challenges for research. PMID- 17412422 TI - Child abuse in the eyes of the beholder: lay perceptions of child sexual and physical abuse. AB - OBJECTIVE: The purpose was to explore the effects of victim and perpetrator gender, type of abuse, and victim-perpetrator relationship on university students' and non-students' perceptions of different kinds of child abuse. METHOD: One hundred and ninety-nine participants (including university students and non-student adults) evaluated each of 24 vignettes (within-subjects design) describing an abusive interaction between a child and an adult. The following four variables were manipulated: the victim's gender, the perpetrator's gender, the type of abuse (physical, relatively mild sexual, or relatively severe sexual), and the perpetrator's relationship to the victim (parent or babysitter). Participants rated each vignette on a number of dimensions: degree of trauma and severity, likelihood of general occurrence and reoccurrence, victim believability, and "repressibility" of the event. RESULTS: Significant interactions emerged on each dimension. For example, sexual abuse (whether mild or severe) was rated as being more traumatic and severe if perpetrated by a parent, but relationship type did not affect perceptions of physical abuse. In addition, significant perpetrator gender by victim gender interactions indicated that homosexual abuse was perceived as more traumatic and repressible than heterosexual abuse, but as less likely to occur; and male participants tended to be more affected by the gender of the perpetrator and abuse type than female participants. CONCLUSION: The results suggest that people have stereotypes about the circumstances and consequences of child abuse. These stereotypes are often, though not always, consistent with existing empirical findings. PMID- 17412423 TI - Evaluation of two immunochromatographic tests (ImmunoCard Toxins A&B, Xpect C. difficile Toxin A&B) and PCR for the detection of Clostridium difficile toxins in faecal samples. PMID- 17412424 TI - Depressed mothers' newborns show less discrimination of other newborns' cry sounds. AB - Newborns' crying in response to the cry of another newborn has been called an empathetic response. The purpose of this study was to determine whether newborns of depressed mothers showed the same response. Newborns of depressed and non depressed mothers were presented with cry sounds of themselves or other infants, and their sucking and heart rate were recorded. The newborns of non-depressed mothers responded to the cry sounds of other infants with reduced sucking and decreased heart rate. In contrast, the newborns of depressed mothers did not show a change in their sucking or heart rate to the cry sounds of other infants. This lesser attentiveness/responsiveness to other infants' cry sounds may predict their later lack of empathy. PMID- 17412425 TI - Molecular beacon genotyping for globoid cell leukodystrophy from hair roots in the twitcher mouse and rhesus macaque. AB - Rapid and accurate genotype determination is ideal for the maintenance of breeding colonies of laboratory animal models of genetic disease. The rhesus macaque and murine (twitcher) models of globoid cell leukodystrophy have a dinucleotide deletion or single nucleotide substitution, respectively, which abolish ceramide beta-galactosidase activity and are authentic models of Krabbe disease. We report a molecular beacon PCR assay for each species which allows unambiguous determination of the genotype in under 4h. The assay works reliably with DNA extracted from hair roots using Chelex-100 in a 20 min, 100 degrees C incubation. We demonstrate that genotyping from hair roots is a preferred alternative to collecting blood or tissue for DNA extraction because it reduces animal distress, uses an inexpensive reagent, and is simpler and faster. Following amplification on a standard thermocycler with a 96-well plate format, these molecular beacon assays can be read on a standard laboratory fluorescent plate reader, eliminating the need to use a real-time thermocycler or to open the plate for subsequent restriction enzyme digestion and gel electrophoresis. The multiplexed ratio of fluorescence from wild-type- and mutant-specific beacons reporting at 560 nm and 535 nm wavelengths is distinct for each genotype. PMID- 17412426 TI - Association of serum cytokine concentrations with psychological symptoms in midlife women. AB - OBJECTIVE: The purpose of the present study was to clarify the association of serum cytokine concentrations, determined using a multiplexed cytokine assay, with psychological symptoms in midlife women. METHODS: Fifty-three peri- and post menopausal women with and without psychological symptoms in Greene's climacteric scale were enrolled in this study. Levels of 17 cytokines in serum samples were measured simultaneously using a multiplexed human cytokine assay. RESULTS: Serum interleukin (IL)-6 concentration in women with psychological symptoms (2.71+/-047 pg/ml) was significantly (p=0.009) higher than that in women without psychological symptoms (0.98+/-0.18 pg/ml). Serum IL-8 concentration in women with psychological symptoms (33.4+/-8.17 pg/ml) was also significantly (p=0.022) higher than that in women without psychological symptoms (7.87+/-1.64 pg/ml). In addition, serum IL-10 concentration in women with psychological symptoms (0.74+/ 0.26 pg/ml) was significantly (p=0.048) higher than that in women without psychological symptoms (0.07+/-0.04 pg/ml). Tumor necrosis factor (TNF)-alpha in serum was detected only in women with psychological symptoms. Serum IL-2 concentration in women with psychological symptoms tended (p=0.066) to be higher than that in women without psychological symptoms. No significant differences were found between levels of other cytokines in women with and without psychological symptoms. CONCLUSION: Psychological stress manifested as climacteric symptoms in midlife women may be associated with increases in serum concentrations of IL-6, IL-8, IL-10, and TNF-alpha. PMID- 17412427 TI - Impaired secondary somatosensory gating in patients with schizophrenia. AB - A large and growing literature has demonstrated a deficit in auditory gating in patients with schizophrenia. Although that deficit has been interpreted as a general gating problem, no deficit has been shown in other sensory modalities. Recent research in our laboratory has examined sensory gating effects in the somatosensory system showing no difference in gating of the primary somatosensory response between patients with schizophrenia and control subjects. This is consistent with recent structural studies showing no cortical structural abnormality in primary somatosensory area in schizophrenia. However, a significant decrease in cortical thickness and gray matter volume loss in secondary somatosensory cortex has recently been reported, suggesting this as a focus for impaired somatosensory gating. Thus, the current study was designed (1) to replicate previous work showing a lack of schizophrenia deficit in primary somatosensory cortex (SI) gating, and (2) to investigate a possible deficit in secondary somatosensory cortex (SII) gating. In a paired-pulse paradigm, dipolar sources were assessed in SI and SII contralateral to unilateral median nerve stimulation. Patients demonstrated no impairment in SI gating, but a robust gating deficit in SII, supporting the presence of cross modal gating deficits in schizophrenia. PMID- 17412428 TI - Identification of Staphylococcus aureus strains negative for enterotoxins A, B and C isolated from bovine mastitis in Mexico. AB - A total of 41 isolates of Staphylococcus aureus obtained from bovine mastitis in 7 different states in Mexico were analyzed by the polymerase chain reaction (PCR) to determine the presence of encoding genes for enterotoxins A, B and C. The oligonucleotides were designed by specific regions of the sea, seb, sec genes. Surprisingly, none of the isolates presented the prospective amplification bands when they were run on agarose gels. On the contrary, reference strains CECT 976 SEA; CECT 5191 SEB; and CECT 4465 SEC showed the prospective amplification products. In order to confirm these results, enterotoxin production A, B, C, D, and E was determined by enzyme linked fluorescent assay (ELFA) using a MiniVIDAS system, on 15 Staphylococcus aureus selected at random from among the 41 isolates. None of the analyzed strains was positive to the test, whereas reference strains enterotoxins producing: CECT 976 SEA; CECT 5191 SEB; CECT 4465 SEC, CECT 4466 SED; CECT 5192 SEE produced concentrations of the toxins detected for this technique. The role of enterotoxins in the pathogenicity of S. aureus in bovine mastitis in Mexico is discussed. PMID- 17412429 TI - Nicotine neuroprotection against nigrostriatal damage: importance of the animal model. AB - Parkinson's disease is a neurodegenerative movement disorder that is characterized by a loss of nigrostriatal dopamine-containing neurons. Unexpectedly, there is a reduced incidence of Parkinson's disease in tobacco users. This finding is important because the identification of the component(s) responsible for this effect could lead to therapeutic strategies to slow down or halt the progression of Parkinson's disease. Results from cell culture models consistently show that nicotine protects against neurotoxicity. However, data from animal models of nigrostriatal damage are conflicting, thus raising questions about a neuroprotective role of nicotine. Accumulating evidence indicates that discrepancies are observed primarily in mouse models of the disease. By contrast, reproducible protection occurs in rat models and in a nonhuman primate parkinsonian model that closely resembles the human disease. These findings highlight the need to use the appropriate animal model and treatment conditions when testing putative neuroprotective agents. PMID- 17412430 TI - Quantitative approaches to analyzing imatinib-treated chronic myeloid leukemia. AB - Progress in understanding the genetic changes that drive tumorigenesis has enabled the development of molecularly targeted anticancer therapy. The first small molecule targeted to a specific protein was imatinib mesylate (Gleevec, STI571), which is used to treat chronic myeloid leukemia (CML). A recent article presents a computational model with which to study the treatment response in CML patients and investigates the effect that imatinib exerts on leukemic stem cells. Here, I discuss insights derived from this study and their implications for imatinib therapy against CML. PMID- 17412431 TI - FXR: a promising target for the metabolic syndrome? AB - The metabolic syndrome is an insulin-resistant state that is characterized by a cluster of cardiovascular risk factors, including abdominal obesity, hyperglycemia, elevated blood pressure and combined dyslipidemia. In this review, we discuss the role of the bile-acid-activated farnesoid X receptor (FXR) in the modulation of the metabolic syndrome. Owing to its regulatory actions in lipid and glucose homeostasis, FXR is a potential pharmacological target. Moreover, the observation that FXR also influences endothelial function and atherosclerosis indicates a regulatory role in the cardiovascular complications that are associated with the metabolic syndrome. The pharmacological activation of FXR leads to a complex response that integrates beneficial actions and potentially undesirable side-effects. Thus, the identification of selective FXR modulators (selective bile acid receptor modulators) is required for the development of compounds that can be used to treat the metabolic syndrome. PMID- 17412432 TI - Lesion of the main olfactory epithelium facilitates maternal behavior in virgin rabbits. AB - Maternal behavior is induced in virgin female rabbits (normally unresponsive to foster pups) by removing the accessory olfactory bulbs. To determine if the main olfactory system (MOS) plays a similar inhibitory role in the present work we investigated the effect of lesioning the olfactory epithelium with a ZnSO(4) spray on the facilitation of maternal behavior in New Zealand white virgin rabbits. Four days after the chemical lesion 40% of females showed behaviors indistinguishable from those of normal mothers, i.e.: rapid entrance into the nest box containing the pups, adoption of a crouching posture over them, acceptance of suckling, and exit from the nest box after ca. 3min. The proportion of females showing these behaviors rose to 70% by day 14 post-lesion. Ovariectomized rabbits sprayed with ZnSO(4) or animals sprayed with NaCl did not behave maternally. ZnSO(4) also provoked a transient reduction in olfactory perception: before the lesion animals from all groups directed significantly more sniffs to a flask containing male urine than to one containing water. This difference was abolished in ZnSO(4)-sprayed females (intact and ovariectomized) for 3-6 days post-lesion and was re-established by 7-9 days. NaCl did not provoke such transitory hyposmia. ZnSO(4) lesions did not provoke malaise in the animals, as determined by food intake and the frequency of scent-marking and ambulation. Results suggest that olfactory cues from the pups are aversive to virgin rabbits and that a transitory reduction in their perception (accompanied by the action of ovarian secretions) is enough to facilitate maternal responsiveness. PMID- 17412433 TI - Neural defects and cardiac arrhythmia in fish larvae following embryonic exposure to 2,2',4,4'-tetrabromodiphenyl ether (PBDE 47). AB - Polybrominated diphenyl ethers (PBDEs) are added to plastics, polyurethane foam, and textiles as a flame retardant. While PBDEs play a key role in reducing loss of human life and property from fires, these flame retardants have become pervasive organic contaminants in the environment and in the tissues of fish, birds, marine mammals, and humans. Levels of PBDEs in wildlife and humans continue to rise, raising concerns about potential ecological and health risks associated with exposure to these chemicals. Nevertheless, there is little currently known about the toxicological effects of PBDE exposure. Here, we examined the developmental toxicity of the PBDE congener 2,2',4,4' tetrabromodiphenyl ether (PBDE 47) using the zebrafish (Danio rerio) as an ontogenetic model. Zebrafish embryos were exposed continuously to dissolved phase PBDE 47 (100-5000 microg/l) beginning 3-5 h post-fertilization (hpf). Fish treated with the highest concentrations of PBDE 47 delayed hatching, had reduced growth post-hatching, and displayed an abnormal dorsal curvature of the body with flexion at the hindbrain. By 96h post-fertilization larvae exposed to PBDE 47 had significant tachycardia, which progressed into atrioventricular block arrhythmias. Microinjection of fluorescent dye into the hindbrain ventricle revealed that cerebrospinal fluid in the neural tube and brain ventricles flowed more slowly in fish larvae exposed to PBDE 47, a likely etiology for the dorsal curvature. Similar, though much less pronounced, developmental toxicity also occurred in larvae exposed to PBDE 47 only for a 20h period during early embryogenesis (3-23 hpf), suggesting that PBDEs incorporated in lipid of the egg are bioavailable and cause toxicity later in life. Taken together, this work indicates that exposure to PBDE 47 can cause morphological abnormalities, impair cardiovascular function and cerebrospinal fluid flow, and provides a tractable starting point for using the zebrafish model to explore molecular mechanisms of PBDE toxicity. PMID- 17412434 TI - A long-term copper exposure on freshwater ecosystem using lotic mesocosms: individual and population responses of three-spined sticklebacks (Gasterosteus aculeatus). AB - Three-spined stickleback (Gasterosteus aculeatus) was used as the highest trophic level predator in an outdoor mesocosm study assessing the effect of environmentally realistic copper concentration (0, 5, 25 and 75 microgL(-1)) over 18 months of continuous exposure. Condition factor, organosomatic indices (HIS, GSI and SSI) as well as copper bioaccumulation in the liver were measured at 15 days, 2, 4, 6, 10, 14 and 18 months after the beginning of the contamination. Population monitoring was realised after 6 and 18 months of contamination, allowing two reproduction periods to be measured. Results showed that condition factor was affected at medium and high copper concentrations and HSI was sporadically affected in all copper exposure, depending on the sex of the fish. GSI did not show any significant differences and SSI was lowered in the medium and high copper levels. Bioaccumulation was significantly different in males and females and fluctuated with season. A negative correlation was observed between copper bioaccumulation in the liver and fish size and a positive correlation with nominal copper concentration in the water was found. There was a negative correlation between condition factor, organosomatic indices and bioaccumulation in the liver. Population monitoring showed a significantly higher fish mean length after 6 months and a higher abundance after 18 months of exposure at the highest copper level. We conclude that indirect effects such as food and habitat availability or lower predation pressure on eggs and juveniles might have led to higher stickleback population abundances at the highest copper level. This highlights the need to study all the trophic levels when monitoring ecosystem health. Considering the population and the individual responses after 18 months of copper exposure, the NOEC for three-spined sticklebacks was 25 microgL(-1) (or 20 microgL(-1) if we consider the average effective concentration), with a LOEC of 75 microgL(-1) (or 57 microgL(-1), AEC). PMID- 17412435 TI - Identification of ribosomal phosphoprotein P0 of Neospora caninum as a potential common vaccine candidate for the control of both neosporosis and toxoplasmosis. AB - The characterization of the cross-reactive antigens of two closely related apicomplexan parasites, Neospora caninum and Toxoplasma gondii, is important to elucidate the common mechanisms of parasite-host interactions. In this context, a gene encoding N. caninum ribosomal phosphoprotein P0 (NcP0) was identified by immunoscreening of a N. caninum tachyzoite cDNA expression library with antisera from mice immunized with T. gondii tachyzoites. The NcP0 was encoded by a gene with open reading frame of 936 bp, which encoded a protein of 311 amino acids. The NcP0 gene existed as a single copy in the genome and was interrupted by a 432 bp intron. The NcP0 showed 94.5% amino acid identity to T. gondii P0 (TgP0). Anti recombinant NcP0 (rNcP0) sera recognized a native parasite protein with a molecular mass of 34 kDa in Western blot analysis. Immunofluorescence analysis showed that the NcP0 was localized to the surface of N. caninum tachyzoites. A purified anti-rNcP0 IgG antibody inhibited the growth of N. caninum and T. gondii in vitro in a concentration-dependent manner. These results indicate that P0 is a cross-reactive antigen between N. caninum and T. gondii and a potential common vaccine candidate to control both parasites. PMID- 17412436 TI - Why is disease progression more rapid in the proximal segments of grafted coronary arteries? AB - Coronary artery segments proximal to the site of graft placement are prone to accelerated atherosclerosis. This has been generally (and somewhat simplistically) attributed to competitive flows between the graft and the native vessel. A more plausible mechanism, linking the rapid induction and progression of atherosclerosis to abnormal flow patterns, and the consequent deleterious alterations in shear stress, is presented here. PMID- 17412437 TI - XL-ECG in athlete's physiological heart hypertrophy: a time saving procedure. PMID- 17412438 TI - Pharmacological washout for the correct evaluation of the head-up tilt testing. AB - Head-up tilt testing is an important tool in the diagnosis of syncope. Several different protocols are in use. We describe the case of a 70-year-old Italian woman admitted to our observation. The patient was in antihypertensive treatment with carvedilol and with a combination of lisinopril and hydrochlorothiazide. A simplified Italian protocol head-up tilt testing was performed. A 4.10 s pause with syncope and a profound hypotension (blood pressure values were 65/50 mm Hg) were observed after 3 min in the provocation phase. Second-degree atrioventricular Block of the 2:1 form, advanced second-degree atrioventricular block and junctional escape rhythm (28 bpm) were observed. A simplified Italian protocol head-up tilt testing was performed after 40 days of withdrawal of carvedilol. A 2 s pause with presyncope and a hypotension (blood pressure values were 80/70 mm Hg) were observed after 2 min in the provocation phase. ECG revealed a bradycardic sinusal rhythm with heart rate of 42 bpm. This case assesses the importance of a pharmacological washout for the correct evaluation of the head-up tilt testing. PMID- 17412439 TI - Perinatal bisphenol A affects the behavior and SRC-1 expression of male pups but does not influence on the thyroid hormone receptors and its responsive gene. AB - Bisphenol A (BPA) has been shown to interfere with thyroid hormone receptors (THRs) and to influence the expression of THR-responsive elements in vivo and in vitro, while some studies reported hyperactivity induced by BPA treatment. In the present study, our purpose was to investigate the effect of BPA exposure on behavioral alteration and its mechanism of action, especially focusing on the thyroid hormone pathway. Significant sexual difference on behaviors was observed in perinatal BPA exposure, as manifested by hyperactivity and impaired spatial learning/memory in male pups after matured. Dams treated with 0.1mg/l BPA showed transient hypothyroidism, while male pups were found to exhibit a transient hyperthyroidism followed by hypothyroidism. Furthermore, significant up-regulated expression levels of mRNA and protein of SRC-1 in the hippocampus were observed in male pups by 0.1mg/l BPA treatment. However the expression of THRalpha/beta and RC3/neurogranin were not affected by BPA treatment. These results indicate that perinatal BPA exposure at a very low level may influence thyroid function and then consequently affects brain development, but at the same time, suggest that thyroid hormone receptor may not be a direct target of BPA action, but instead, another factor may be involved in this action. PMID- 17412440 TI - Occurrence of Fusarium species and trichothecenes in Nigerian maize. AB - A total of 180 maize samples meant for human consumption from four maize producing states of southwestern Nigeria were screened for twelve major Fusarium mycotoxins (trichothecenes). Mycological examination of the samples showed that Fusarium verticillioides was the most commonly isolated fungi (71%), followed by F. sporotrichioides (64%), F. graminearum (32%), F. pallidoroseum (15%), F. compactum (12%), F. equiseti (9%), F. acuminatum (8%), F. subglutinans (4%) and F. oxysporum (1%). The trichothecenes include deoxynivalenol (DON), 3, mono acetyldeoxynivalenol (3-AcDON), 15, mono-acetyldeoxynivalenol (15-AcDON), nivalenol (NIV), HT-2 toxin (HT-2), neosolaniol (NEO), T-2 toxin (T-2), T-2 tetraol and T-2 triol, diacetoxyscirpenol (DAS), MAS-monoacetoxyscirpenol (MAS) and fusarenone-X. Quantification was by high performance liquid chromatography coupled with mass spectroscopy (HPLC/MS); the detection limits for each of the mycotoxins varied between 20 and 200 microg kg(-1). Sixty six samples (36.3%) were contaminated with trichothecenes, DON (mean: 226.2 microg kg(-1); range: 9.6 745.1 microg kg(-1)), 3-AcDON (mean: 17.3 microg kg(-1); range: 0.7-72.4 microg kg(-1)) and DAS (mean: 16.0 microg kg(-1); range: 1.0-51.0 microg kg(-1)) were detected in 22%, 17% and 9% of total samples respectively. There were no 15 AcDON, NIV, HT-2, NEO, T-2, T-2 tetraol, T-2 triol, MAS and fusarenone-X detected. This is the first comprehensive report about the natural occurrence of DON, AcDON and DAS in maize for direct human consumption in Nigeria. PMID- 17412441 TI - Identification and quantification of N alpha-acetylated Y. pestis fusion protein F1-V expressed in Escherichia coli using LCMS E. AB - N-terminal acetylation in E coli is a rare event catalyzed by three known N acetyl-transferases (NATs), each having a specific ribosomal protein substrate. Multiple, gram-scale lots of recombinant F1-V, a fusion protein constructed from Y. Pestis antigens, were expressed and purified from a single stably transformed E. coli cell bank. A variant form of F1-V with mass increased by 42-43 Da was detected in all purified lots by electrospray orthogonal acceleration time-of flight mass spectrometry (MS). Peptide mapping LCMS localized the increased mass to an N-terminal Lys-C peptide, residues 1-24, and defined it as +42.0308+/ 0.0231 Da using a LockSpray exact mass feature and a leucine enkaphalin mass standard. Sequencing of the variant 1-24 peptide by LCMS and high-energy collision induced dissociation (LCMS(E)) further localized the modification to the amino terminal tri-peptide ADL and identified the modification as N(alpha) acetylation. The average content of N(alpha)-acetylated F1-V in five lots was 24.7+/-2.6% indicating that a stable acetylation activity for F1-V was established in the E. coli expression system. Alignment of the F1-V N-terminal sequence with those of other known N(alpha)-acetylated ectopic proteins expressed in E. coli reveals a substrate motif analogous to the eukaryote NatA' acetylation pathway and distinct from endogenous E. coli NAT substrates. PMID- 17412442 TI - Attenuated dengue 2 viruses with deletions in capsid protein derived from an infectious full-length cDNA clone. AB - A full-length cDNA clone (pD212) of dengue virus type 2 isolated in China (DEN2 43) was constructed. Based on this, we constructed several mutants with deletions in capsid protein C using fusion PCR. These deletions removed part or almost all of the internal stretch of hydrophobic amino acid residues that is probably involved in virion assembly. We thus obtained viable mutant viruses. The propagation capacity of the mutant viruses in cell culture was impaired in parallel with the increasing size of the deletion, and the infectivity of mutant C(Delta42-59), from which all of helix III of capsid protein C was removed, was completely abolished. More importantly, the mutant viruses were highly attenuated in suckling mice but induced high levels of antibodies in adult mice. This study indicates that the structural and functional flexibility of capsid protein C make it a candidate target for the attenuation of dengue virus, which could open a promising new avenue for the development of live attenuated dengue vaccines. PMID- 17412443 TI - Preparation, in vitro and in vivo evaluation of stomach-specific metronidazole loaded alginate beads as local anti-Helicobacter pylori therapy. AB - Metronidazole (MZ), a common antibacterial drug used in treatment of H. pylori, was prepared in chitosan-treated alginate beads by the ionotropic gelation method. A (3x2x2) factorially designed experiment was used in which 3 viscosity imparting polymers namely, methyl cellulose, carbopol 934P and kappa-carrageenan, 2 concentrations (0.2 and 0.4% w/v) of chitosan as encapsulating polymer and 2 concentrations (2.5 and 5% w/w) of the low density magnesium stearate as a floating aid were tested. The drug entrapment efficiency (%), the percent of floating beads and the time for 80% of the drug to be released (T(80%)) were the responses evaluated. The bead formula containing 0.5% kappa-carrageenan, 0.4% chitosan and 5% magnesium stearate showed immediate buoyancy, optimum drug entrapment efficiency and extended drug release. The histopathological examination of mice stomachs and in vivo H. pylori clearance tests were carried out by orally administering MZ floating alginate beads or MZ suspension, to H. pylori infected mice under fed conditions as a single daily dose for 3 successive days in different doses 5, 10, 15 and 20 mg/kg. The histopathological examination showed that groups receiving MZ in the form of floating alginate beads at doses 10, 15 and 20 mg/kg were better than the corresponding suspension form, regarding eradication of H. pylori infection. The in vivo H. pylori clearance tests showed that MZ floating beads with a dose of 15 mg/kg provided 100% clearance rate whereas the MZ suspension with a dose of 20 mg/kg gave only 33.33%. PMID- 17412444 TI - Influence of wettability and surface activity on release behavior of hydrophilic substances from lipid matrices. AB - The aim of this study was to investigate the role of matrix and drug properties on controlled release from triglyceride matrices. Mini-cylinders of 2 mm diameter, 2.2 mm height and 7 mg weight were produced by compression of lipid powder obtained by using a polyethylene glycol (PEG) co-lyophilization method for the model substances lysozyme and FITC-dextran (Mw 4000 Da). Lysozyme was released with decreasing velocity from glyceryl trilaurate, -myristate, palmitate and -stearate for more than 14 months. Release correlated well with triglyceride lipophilicity defined by the chain length of the fatty acids. Contact angle measurements and the analysis of buffer penetration visualized by confocal microscopy emphasized the role of matrix wettability as a prerequisite for release. A comparison with FITC-dextran revealed that the protein itself enhances matrix wettability and hence its release due to its surface active properties. FITC-dextran remained trapped within the matrix only to be released at lower compression force or after the addition of surfactant. Protein added externally to the release buffer at 0.1% (w/v) was not efficient in lowering the contact angle and increasing the release rate of FITC-dextran. Tween 20 and 81 could be used in different concentrations (0.1, 0.01 and 0.001% (w/v)) to alter lysozyme and FITC-dextran release profiles: resulting release rates showed a close dependence on the contact angle of the respective release medium and triglyceride matrix material. However, both Tweens seem to act not only by reducing the release medium contact angle but also by moderately affecting interparticulate adhesion of the matrix material. PMID- 17412445 TI - Continuous passive motion compared with intermittent mobilization after total knee arthroplasty. Elaboration of French clinical practice guidelines. AB - OBJECTIVE: To develop clinical practice guidelines concerning the use of continuous passive motion (CPM) compared with intermittent mobilization after total knee arthroplasty (TKA). METHOD: We used the SOFMER (French Physical Medicine and Rehabilitation Society) methodology, combining systematic literature review and collection of everyday clinical practice concerning postoperative rehabilitation techniques and external review by a multidisciplinary expert panel, to develop the guidelines. RESULTS: The literature contains no evidence of the advantages of CPM over other techniques of mobilization, although CPM could be adjuvant therapy used to accelerate short-term recovery. However, in France, CPM remains widely used after TKA, both in orthopedic surgery units and in physical medicine and rehabilitation services. CONCLUSION: Good methodological quality studies are needed to assess different CPM modalities and compare them to alternative intermittent mobilization techniques, particularly those with therapy starting from a flexed position. PMID- 17412446 TI - Diabetes and exercise-induced hemodynamic responses. PMID- 17412447 TI - Methodology of superiority vs. equivalence trials and non-inferiority trials. AB - The randomized clinical trial (RCT) is generally accepted as the best method of comparing effects of therapies. Most often the aim of an RCT is to show that a new therapy is superior to an established therapy or placebo, i.e. they are planned and performed as superiority trials. Sometimes the aim of an RCT is just to show that a new therapy is not superior but equivalent to or not inferior to an established therapy, i.e. they are planned and performed as equivalence trials or non-inferiority trials. Since the types of trials have different aims, they differ significantly in various methodological aspects. The awareness of the methodological differences is generally quite limited. This paper reviews the methodology of these types of trials with special reference to differences in respect to planning, performance, analysis and reporting of the trial. In this context the relevant basal statistical concepts are reviewed. Some of the important points are illustrated by examples. PMID- 17412448 TI - Early ribavirin pharmacokinetics, HCV RNA and alanine aminotransferase kinetics in HIV/HCV co-infected patients during treatment with pegylated interferon and ribavirin. AB - BACKGROUND/AIMS: We evaluated whether early ribavirin pharmacokinetics differ comparing hepatitis C/human immunodeficiency virus coinfected sustained virological responders and nonresponders. METHODS: Twenty-four treatment-naive coinfected patients received pegylated-interferon alfa-2b (12 kDa) (1.5 microg/kg) once weekly plus daily ribavirin (13.6 mg/kg/d) for up to 48 weeks. Serum HCV RNA, serum alanine aminotransferase, and plasma ribavirin levels were measured frequently during the first 16 days of therapy and monthly thereafter. RESULTS: Six patients were sustained responders. During the first 4 weeks of treatment, median plasma ribavirin levels and area under the ribavirin curve were significantly lower (p<0.0001 and p<0.01, respectively) in sustained responders compared with nonresponders. Compared to ribavirin levels at weeks 2 and 4, ribavirin levels in sustained responders continued to increase significantly until week 8 (p<0.02). At week 4, hemoglobin declines were significantly (p=0.002) greater in sustained responders than nonresponders. At week 1, serum HCV RNA levels and changes in alanine aminotransferase levels relative to baseline could identify likely responders better than plasma ribavirin levels. CONCLUSIONS: We conjecture that intracellular ribavirin accumulation may be enhanced early in treatment in coinfected sustained responders, although this hypothesis should be investigated further. At week 1, serum HCV RNA and changes in alanine aminotransferase levels relative to baseline might identify likely responders. PMID- 17412449 TI - A recognition-free mechanism for reliable rejection of brood parasites. AB - Hosts often discard eggs of avian brood parasites, whereas parasitic chicks are typically accepted. This can be explained theoretically by fitness losses associated with adults learning to recognize parasitic young and mistakenly rejecting their own young. A new experimental study confirms that rejection of parasitic chicks, without relying on memory to discriminate between foreign and own young, is a feasible and potentially cost-free mechanism used by reed warblers to reject common cuckoo chicks. By abandoning broods that are in the nest longer than is typical for their own young, parents can reliably reject parasite nestlings and reduce fitness losses owing to having to care for demanding parasitic young. Discrimination without recognition has important implications for the realized trajectories of host-parasite coevolutionary arms races. PMID- 17412450 TI - Determining conditional stability constants for Pb complexation by carboxymethyl beta-cyclodextrin (CMCD). AB - Carboxymethyl-beta-cyclodextrin (CMCD) has been proposed for remediation of metal contaminated sediments. This research presents stability constants for CMCD-lead complexes, and demonstrates a rigorous methodology for estimating stability constants for metal-complexing agents. The conditional stability constant for the lead-CMCD aqueous complex was determined to be 10(5.18) with the 95% confidence interval ranging from 10(5.14) to 10(5.22). The best fit for experimental data was made by assuming a reaction between divalent CMCD(2-) and Pb(2+) and using the WATEQ activity coefficient formulation. The optimized value was derived from experimental data with the geochemical model PHREEQC coupled to UCODE_2005, a parameter optimization program. Like FITEQL, UCODE has a built-in option to optimize parameter values by minimizing the weighted sum of squared residuals (WSSR). However, our approach not only allows rapid, automatic optimization of the stability constant, but also allows determination of uncertainties in estimated parameter values and statistical analysis to assess the appropriateness of the conceptual model. The automation of the process allows testing of multiple conceptual models and the final values produced are internally consistent with the PHREEQC database. In this case five different conceptual models to describe the metal complexation and protonation reactions of CMCD were considered. PMID- 17412451 TI - Nephrogenic systemic fibrosis: a clinicopathologic study of six cases. AB - BACKGROUND: Nephrogenic systemic fibrosis is a rare fibrosing condition that occurs in patients with renal insufficiency. While its histologic characteristics have been well described, the etiology and pathogenesis have not been fully characterized. Several recent studies support the theory that gadolinium-based contrast agents play a causative role in the development of the disease. Erythropoietin therapy and endothelial damage from surgical procedures have also been suggested as potential contributing factors. OBJECTIVE: This study attempts to help contribute to the understanding of this novel disorder. METHODS: We performed a retrospective chart review of 6 patients diagnosed with nephrogenic systemic fibrosis at our institution. Emphasis was placed on identification of potential putative etiologic agents including gadolinium, erythropoietin therapy, and previous surgical procedures. RESULTS: All patients had documented exposure to gadolinium-based contrast agents. Three of the 6 patients were treated with erythropoietin, and all patients underwent a previous surgical procedure. LIMITATIONS: This study is limited by its small size; therefore, the findings and results may not be applicable to all patients with this disorder. CONCLUSION: Our data suggest that gadolinium plays a primary role in nephrogenic systemic fibrosis and that prior surgery may be a contributory factor. PMID- 17412452 TI - Circumscribed acral hypokeratosis. AB - BACKGROUND: Circumscribed acral hypokeratosis (CAH) is an idiopathic condition that typically presents as an acquired, solitary, asymptomatic, well-defined, depressed, flat-based deformity, with a slightly raised border on the palm or, rarely, the sole. Histologically, the lesional epidermis is depressed with a characteristic, abrupt, hyperkeratotic, slightly-raised ridge at the transition from normal skin. OBJECTIVE: We sought to present 3 additional cases of CAH. METHODS: A review of 3 cases of CAH was performed. Liquid phase polymerase chain reaction (PCR) was conducted to evaluate for human papillomavirus (HPV). RESULTS: Three cases of CAH were reviewed. One patient had a history of a burn at the site prior to developing the disorder while another patient, as a child, had a history of verruca plantaris in the same location. Lesions were solitary, involving the palm or sole, in 2 cases and in one case they were multiple involving both the palms and the soles. HPV testing detected HPV type 6 in the lesion of one patient who previously was treated for warts in the same location. Topical fluorouracil, calcipotriol ointment under occlusion, and clobetasol ointment under occlusion were unsuccessful in one patient. LIMITATIONS: In case 2, we were pathology consultants and unable to evaluate the clinical appearance of the lesion. CONCLUSIONS: CAH may involve palms and/or soles. Lesions may be solitary or multiple, and vary widely in size. We believe that CAH most likely represents a reaction pattern developing in response to various stimuli, including trauma, HPV, or both. PMID- 17412453 TI - Merkel cells. AB - Merkel cells are post-mitotic cells scattered throughout the epidermis of vertebrates. They are particularly interesting because of the close connections that they develop with sensory nerve endings and the number of peptides they can secrete. These features suggest that they may make an important contribution to skin homeostasis and cutaneous nerve development. However, these cells remain mysterious because they are difficult to study. They have not been successfully cultured and cannot be isolated, severely hampering molecular biology and functional analysis. Merkel cells probably originate in the neural crest of avians and mammalians, and their "spontaneous" appearance in the epidermis may be caused by a neuron-independent epidermal differentiation process. Their functions are still unclear: they take part in mechanoreception or at least interact with neurons, but little is known about their interactions with other epidermal cells. This review provides a new look at these least-known cells of the skin. The numerous peptides they synthesize and release may allow them to communicate with many cells other than neurons, and it is plausible that Merkel cells play a key role in skin physiology and physiopathology. PMID- 17412454 TI - Water content and other aspects of brittle versus normal fingernails. AB - BACKGROUND: Previous authors have claimed that dehydration of the nail plate causes brittle nails. Some experts claim that normal nails contain 18% water, and brittle nails contain less than 16%. OBJECTIVE: We sought to test the hypothesis that brittle nails contain 2% less water than normal nails. We also examined the relationship between a number of health and behavioral variables and brittle nails. METHODS: In all, 102 participants with either brittle or normal nails had two nails clipped and then analyzed for water content by a blinded investigator in the laboratory. Participants filled out a detailed questionnaire designed to reveal information about health and behavior. RESULTS: The mean water content for normal nails was 11.90% and for brittle nails was 12.48%. There was no statistically significant difference between the two groups. The odds of having brittle nails was 3.23 times greater among participants who received a professional manicure (95% confidence interval 1.21, 8.59). The frequency of professional manicures was associated with the likelihood of having brittle nails. Frequency of hand moisturizer use was significantly associated with nail brittleness (95% confidence interval 1.35, 32.10). Family history was significantly associated with the likelihood of having brittle nails (95% confidence interval 1.65, 21.11). LIMITATIONS: Analyzing nails from living participants is limiting because samples can only be collected from the distal unattached nail plate. A small subanalysis was performed and showed that the nails were losing water between the time of clipping and laboratory analysis. Therefore, our water percentage results may not be representative of in vivo nail plate water contents. CONCLUSIONS: There was no significant difference in water content of brittle compared with normal nails. PMID- 17412455 TI - Effects of statins on alpha7 nicotinic receptor, cholinesterase and alpha-form of secreted amyloid precursor peptide in SH-SY5Y cells. AB - In order to reveal the neuroprotective effects of statins that could be of interest for the prevention and treatment of Alzheimer's disease (AD), we investigated the expression of nicotinic acetylcholine receptors (nAChRs) detected by RT-PCR, the activity of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) by colorimetric determination, and the levels of the alpha-form of secreted beta-amyloid precursor protein (alphaAPPs) by Western blotting in neuroblastoma (SH-SY5Y) cells exposed to lovastatin, atorvastatin, rosuvastatin and simvastatin, respectively. The results indicated that all statins studied, both lipophilic and hydrophilic, induced high expression of alpha7 nAChR, decreased cholinesterase activities, and increased alphaAPPs, which suggests that statins might play important neuroprotective roles in AD treatment. PMID- 17412456 TI - Synthesis and cytotoxicity studies of new dimethylamino-functionalised and aryl substituted titanocene anti-cancer agents. AB - From the carbolithiation of 6-N,N-dimethylamino fulvene (3a) and different lithiated aryl species [p-N,N-dimethylanilinyl lithium, p-anisyl lithium and 4 lithio-benzo[1.3]dioxole (2a-c)], the corresponding lithium cyclopentadienide intermediates 4a-c were formed. These three lithiated intermediates underwent a transmetallation reaction with TiCl4 resulting in dimethylamino-functionalised and aryl-substituted titanocenes 5a-c. When these titanocenes were tested against LLC-PK cells, the IC50 values obtained were of 54, 45 and 26microM for titanocenes 5a, b and c, respectively. The most cytotoxic titanocene in this paper, 5c is approximately 10 times less cytotoxic than cis-platin, which showed an IC50 value of 3.3microM, when tested on the LLC-PK cell line, but approximately 100 times better than titanocene dichloride. PMID- 17412457 TI - Challenges in teaching undergraduate psychology courses to nursing students. PMID- 17412458 TI - The interaction between policy and education using stroke as an example. AB - This paper discusses the interaction between healthcare policy at the European, UK and Scottish levels and the funding of education that underpins specific health policy priorities. Stroke is used throughout to illustrate the relationship between a designated European and UK health priority and the translation of that priority into clinical delivery. The necessity to build a responsive and sustainable culture to address the healthcare education that underpins changing healthcare policies is emphasized. PMID- 17412459 TI - Nursing and the art of radical critique. AB - This paper argues that more critique is required in our nursing journals. I begin by distinguishing between conservative 'old' critique which functions to maintain the status quo in the academic discipline of nursing, and radical 'new' critique which challenges it and pushes at its boundaries. I then identify three reasons why I believe so little radical critique is published in nursing journals, and illustrate each with examples from my own experience. Firstly, there is an assumption that peer reviewers are fulfilling this critical function, whereas I have argued that peer review should be concerned only with procedural matters and not with 'new' or radical critique, which in any case should be in the public domain for it to be effective and influential. Secondly, radical critique is frequently mistaken for ad hominem attack, causing reluctance amongst writers, reviewers and editors to see it made public. And thirdly, radical critique lies outside of the dominant academic discourse and is therefore itself subject to the very same conservative and repressive attitudes against which it is poised. Nurse academics would do well to look to other disciplines such as philosophy and the arts for examples of radical critique and of the ways in which journal editors and contributors strive to keep academic debate and discourse alive. PMID- 17412460 TI - Effects of fungal N- and O-linked mannosylation on the immunogenicity of model vaccines. AB - Targeting dendritic cell mannose receptors by mannosylating antigens represents a promising vaccination strategy. Using the model antigen ovalbumin (OVA) expressed recombinantly in bacterial and yeast vectors, we have previously demonstrated fungal mannosylation enhances antigen immunogenicity in the context of CD4(+) T cell responses. However, because protection against many tumors and pathogens is thought to require MHC class I-restricted T cell responses, the capacity of differentially mannosylated OVA antigens to induce antigen-specific CD8(+) T cell proliferation was determined. We found that mannosylated yeast-derived OVA antigens were more potent than their unmannosylated counterparts at inducing antigen-specific T cell proliferation. However, the type of mannosylation was critical as addition of extensive O-linked mannosylation increased lymphoproliferative responses while the presence of N-linked mannosylation was associated with decreased responses. Mannosylated OVA failed to stimulate TNF alpha and IL-12 production from dendritic cells. These data suggest that vaccines incorporating mannosylation must take into account how the mannose groups are linked to the core antigen and may need to include an adjuvant to stimulate cytokine production. PMID- 17412461 TI - Comparison of neutralising antibody assays for detection of antibody to influenza A/H3N2 viruses: an international collaborative study. AB - A study was performed to investigate the reproducibility of haemagglutinin inhibition (HI) and virus neutralising (VN) assays for detection of anti influenza antibody. Participants in 11 laboratories from eight countries measured antibody to egg-grown A/Japan/434/2003, cell-grown A/Japan/434/2003 and A/Panama/2007/99 (H3N2) viruses in 18 human and two post-infection ferret sera. There was significant intra-laboratory assay variability for VN compared to HI. For replicate assays within laboratories, 14/410 (3%) and 130/631 (21%) titres differed by >2-fold (p<0.0001), and 0/410 (0%) and 35/631 (6%) titres differed by >5-fold (p<0.0001) by HI and VN, respectively. Although both assays showed inter laboratory variation, VN assays were significantly more variable than HI. Median geometric coefficients of variation (GCV) for VN assays with each virus were 256%, 323% and 359% compared to 138%, 155% and 261% with HI. A serum standard improved inter-laboratory agreement and reduced median GCVs. This study raises concern about comparability of serology results from H5N1 vaccine trials and it is proposed that an International Standard for influenza H5N1 antibody is developed. PMID- 17412463 TI - Use of a polyvinyl acetyl sponge (Merocel) nasal pack to prevent alar necrosis during prolonged nasal intubation. PMID- 17412462 TI - Ability of SPI2 mutant of S. typhi to effectively induce antibody responses to the mucosal antigen enterotoxigenic E. coli heat labile toxin B subunit after oral delivery to humans. AB - We have evaluated an oral vaccine based on an Salmonella enteric serovar typhi (S. typhi) Ty2 derivative TSB7 harboring deletion mutations in ssaV (SPI-2) and aroC together with a chromosomally integrated copy of eltB encoding the B subunit of enterotoxigenic Escherichia coli heat labile toxin (LT-B) in volunteers. Two oral doses of 10(8) or 10(9)CFU were administered to two groups of volunteers and both doses were well tolerated, with no vaccinemia, and only transient stool shedding. Immune responses to LT-B and S. typhi lipopolysaccharide were demonstrated in 67 and 97% of subjects, respectively, without evidence of anti carrier immunity preventing boosting of LT-B responses in many cases. Further development of this salmonella-based (spi-VEC) system for oral delivery of heterologous antigens appears warranted. PMID- 17412464 TI - Asymmetrical shoulder kinematics in children with brachial plexus birth palsy. AB - BACKGROUND: Shoulder movement patterns differ between limbs of children with unilateral brachial plexus birth palsy. To better understand the interlimb differences we examined the glenohumeral and scapulothoracic joint contributions to arm elevation. METHODS: Sixteen children with brachial plexus birth palsy, 4 12 years of age participated. Shoulder 3D kinematic data were collected using a magnetic tracking device during arm elevation with the involved and non-involved limbs for three trials each at a fixed rate. Based on maximum arm elevation in the involved limb the children were divided into two groups: group one 75 degrees . FINDINGS: During arm elevation from 15 to 75 degrees the involved limb of group one displayed lower glenohumeral joint excursion than the non-involved and both limbs of group two. Scapular upward rotation was higher in the involved limb of both groups. For group one, the glenohumeral:scapulothoracic ratio for 15-75 degrees arm elevation was lower in the involved (0.6:1) than the non-involved (2.2:1) limb and both limbs of group two: involved (1.7:1); non-involved (1.9:1). During 15-135 degrees arm elevation for group two, the glenohumeral:scapulothoracic ratio was more similar between limbs: involved (1.5:1) and non-involved (2:1). INTERPRETATION: The scapulothoracic joint made a greater contribution to arm elevation than the glenohumeral joint only in the involved limb of group one, altering the scapulohumeral rhythm. Musculoskeletal and neural factors may account for the group and limb differences. Routine 3D kinematic analysis of shoulder joint rotation may aid treatment planning and better quantify outcomes in this group. PMID- 17412465 TI - Assessing the risk caused by ground level ozone to European forest trees: a case study in pine, beech and oak across different climate regions. AB - Two different indices have been proposed for estimation of the risk caused to forest trees across Europe by ground-level ozone, (i) the concentration based AOT40 index (Accumulated Over a Threshold of 40 ppb) and (ii) the recently developed flux based AFstY index (Accumulated stomatal Flux above a flux threshold Y). This paper compares the AOT40 and AFstY indices for three forest trees species at different locations in Europe. The AFstY index is estimated using the DO(3)SE (Deposition of Ozone and Stomatal Exchange) model parameterized for Scots pine (Pinus sylvestris), beech (Fagus sylvatica) and holm oak (Quercus ilex). The results show a large difference in the perceived O(3) risk when using AOT40 and AFstY indices both between species and regions. The AOT40 index shows a strong north-south gradient across Europe, whereas there is little difference between regions in the modelled values of AFstY. There are significant differences in modelled AFstY between species, which are predominantly determined by differences in the timing and length of the growing season, the periods during which soil moisture deficit limits stomatal conductance, and adaptation to soil moisture stress. This emphasizes the importance of defining species-specific flux response variables to obtain a more accurate quantification of O(3) risk. PMID- 17412466 TI - Effect of temperature, gas phase composition, pH and microbial activity on As, Zn, Pb and Cd mobility in selected soils in the Ebro and Meuse Basins in the context of global change. AB - This study estimates the effect of environmental parameters on the mobility of four inorganic contaminants (As, Zn, Pb and Cd) in soils from three areas in the Ebro and Meuse River basins, within the context of global change. An experimental method, applicable to various soil systems, is used to measure the effect of four global-change-sensitive parameters (temperature, gas phase composition, pH and microbial activity). The aqueous phase of batch incubations was sampled regularly to monitor toxic element concentrations in water. Statistical processing enabled discrimination of the most relevant variations in dissolved concentrations measured at different incubation times and under different experimental conditions. Gas phase composition was identified as the most sensitive parameter for toxic element solubilization. This study confirms that total soil concentrations of inorganic pollutants are irrelevant when assessing the hazard for ecosystems or water resource quality. PMID- 17412467 TI - Use of ethylenediurea (EDU) to ameliorate ozone effects on purple coneflower (Echinacea purpurea). AB - Purple coneflower plants (Echinacea purpurea) were placed into open-top chambers (OTCs) for 6 and 12 weeks in 2003 and 2004, respectively, and exposed to charcoal filtered air (CF) or twice-ambient (2x) ozone (O3) in 2003, and to CF, 2x or non filtered (NF), ambient air in 2004. Plants were treated with ethylenediurea (EDU) weekly as a foliar spray. Foliar symptoms were observed in >95% of the plants in 2x-treated OTCs in both years. Above-ground biomass was not affected by 2x treatments in 2003, but root and total-plant biomass decreased in 2004. As a result of higher concentrations of select cell wall constituents (% ADF, NDF and lignin) nutritive quality was lower for plants exposed to 2x-O3 in 2003 and 2004 (26% and 17%, respectively). Significant EDU x O3 interactions for concentrations of cell wall constituents in 2003 indicated that EDU ameliorated O3 effects on nutritive quality. Interactions observed in 2004 were inconsistent. PMID- 17412468 TI - Export of toxic chemicals - a review of the case of uncontrolled electronic-waste recycling. AB - This paper reviews the concentrations of persistent organic pollutants such as flame retardants (PBDEs), dioxins/furans (PCDD/Fs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals/metalloid concentrations of different environmental media at Guiyu, a traditional rice growing village located in southeastern Guangdong Province (PR China), which has turned into an intensive electronic-waste (e-waste) recycling site. Incomplete combustion of e-waste in open air and dumping of processed materials are the major sources of various toxic chemicals. By comparing with existing data available in other areas and also guidelines adopted in different countries, it is obvious that the environment is highly contaminated by these toxic chemicals derived from the recycling processes. For example, the monthly concentration of the sum of 22 PBDE congeners contained in PM(2.5) (16.8ngm(-3)) of air samples at Guiyu was 100 times higher than published data. In order to safeguard the environment and human health, detailed investigations are urgently needed, especially on tracking the exposure pathways of different toxic chemicals which may affect the workers and local residents especially mothers, infants and children. PMID- 17412469 TI - Amelioration of alkali soil using flue gas desulfurization byproducts: productivity and environmental quality. AB - In this study, flue gas desulfurization (FGD) byproducts are used to ameliorate alkali soil. The average application rates for soils with low exchangeable sodium percentage (ESP), mid ESP, and high ESP are 20.9, 30.6, and 59.3 Mg ha(-1), respectively. The experimental results obtained for 3 consecutive years reveal that the emergence ratios and yields of the crops were 1.1-7.6 times and 1.1-13.9 times those of the untreated control, respectively. The concentrations of Cr, Pb, Cd, As, and Hg in the treated soils are far below the background values stipulated by the Environmental Quality Standard for Soils (GB15618-1995). Their concentrations in the seeds of corn and alfalfa grown in the treated soils are far below the tolerance limits regulated by National Food Standards of China. The results of this research demonstrate that the amelioration of alkali soils using FGD byproducts is promising. PMID- 17412470 TI - Support for selection of a methamphetamine cleanup standard in Colorado. AB - Methamphetamine production for illicit use occurs in makeshift labs and is associated with the release of numerous chemicals, including methamphetamine residues. These methamphetamine residues may pose a health risk to residents who reoccupy these structures after property seizures. Several states have established technology-based cleanup standards for methamphetamine, but none have examined the health-protectiveness of these standards. In response to Colorado House Bill 04-1182, exposure intakes correlated with three technology-based standards were calculated for various groups of individuals. Intakes were assessed for a 1-year-old infant, 6-year-old child, and a female of childbearing age. Exposure intakes were compared to toxicity reference values developed from developmental endpoints following methamphetamine exposure from the available literature. Uncertainty factors were applied to the lowest adverse effect levels observed in these studies to arrive at the toxicity reference values. These reference values were greater than the calculated intakes from each proposed technology standard, suggesting that all of the proposed standards would be protective of human health exposure. The cost and practicality of attaining each of the proposed standards was also factored into the decision making process. In their final regulation (6 CCR 1014-3), the CDPHE selected 0.5 microg/100 cm(2) as the final cleanup standard for methamphetamine residues. PMID- 17412471 TI - Girl, woman, lover, mother: towards a new understanding of child prostitution among young Devadasis in rural Karnataka, India. AB - The emotive issue of child prostitution is at the heart of international debates over 'trafficking' in women and girls, the "new slave trade", and how these phenomena are linked with globalization, sex tourism, and expanding transnational economies. However, young sex workers, particularly those in the 'third world', are often represented through tropes of victimization, poverty, and "backwards" cultural traditions, constructions that rarely capture the complexity of the girls' experiences and the role that prostitution plays in their lives. Based on ethnographic fieldwork with girls and young women who are part of the Devadasi (servant/slave of the God) system of sex work in India, this paper introduces an alternative example of child prostitution. Demonstrating the ways in which this practice is socially, economically, and culturally embedded in certain regions of rural south India underlies this new perspective. I argue that this embeddedness works to create, inform, and give meaning to these girls as they grow up in this particular context, not to isolate and produce totally different experiences of family, gender identity, and moral character as popular accounts of child prostitution contend. Data pertaining to socialization, 'positive' aspects of being a young sex worker in this context, political economy, HIV/AIDS, and changes in the Devadasi tradition are used to support my position. Taken together, this alternative example presents a more complex understanding of the micro- and macro-forces that impact child prostitution as well as the many factors that affect the girls' ideas of what they do and who they are as people, not just sex workers. PMID- 17412472 TI - Engagement in a cohesive group and higher-level functional capacity in older adults in Japan: a case of the Mujin. AB - The Mujin is a traditional Japanese rotating saving and credit association (RoSCA) that provided financial aid for the lower-middle class in Japan until the postwar reconstruction period. The traditional Mujin has disappeared from most regions, and the surviving Mujin provides more of a social networking than a financial function for its members. We investigated the association between level of participation in the Mujin and the capacity for independent living in older adults. Baseline data of an ongoing cohort study of 581 self-sufficient older adults were analyzed. The Tokyo Metropolitan Institute of Gerontology Index of Competence was used to measure higher-level functional capacity, and the level of engagement in the Mujin was determined by a score derived by factor analysis. We found that a higher level of engagement in the Mujin was associated with greater functional capacity, especially social role performance, the highest-level function. We conclude that the Mujin provides a venue for community interaction that serves to increase the social capital of the community and may have psychosocial benefits for its members due to strong membership ties and mutual trust. Further investigation of the causal relationship between participation in the Mujin and functional capacity, and the psychosocial effects of similar financial mutual-aid systems in other social contexts is warranted. PMID- 17412473 TI - Efficacy of flupentixol and risperidone in chronic schizophrenia with predominantly negative symptoms. AB - The study investigated the non-inferiority of flupentixol compared to risperidone in the treatment of negative symptoms. In addition, the effects of flupentixol on mood and cognitive symptoms were explored. In a randomized, double-blind multicenter study, 144 non-acute schizophrenia patients with predominant negative symptoms were treated with a flexible dose of either flupentixol (4-12 mg/d) or risperidone (2-6 mg/d) for up to 25 weeks. In addition to a non-inferiority analysis, a principal component analysis (PCA) of the PANSS was performed post hoc. Regarding negative symptoms, flupentixol proved to be non-inferior to risperidone. Both drugs improved depressed mood with effect sizes favoring flupentixol. PCA suggested a five-factor structure. Effect sizes for the cognitive factor were up to 0.74 for flupentixol and up to 0.80 for risperidone. EPS scores were rather low and Parkinsonism improved in both groups, but anticholinergic drugs were prescribed significantly more frequently in the flupentixol group, which generally showed significantly more adverse events. Results indicate that the 1st generation antipsychotic flupentixol improves negative, affective and cognitive symptoms in chronic schizophrenia comparable to risperidone. Further studies should confirm the latter using neuropsychological performance tests and should investigate whether tolerability improves with a markedly lower dose range. PMID- 17412474 TI - Exposure of the population of the Slovak Republic to dietary polychlorinated biphenyls. AB - The aim of this paper has been to assess the exposure of the Slovak Republic population to polychlorinated biphenyls from food within 1994-2004. A total of 61167 samples of food of animal origin, including fats, were analyzed for the content of six PCB congeners (the sum of: PCB-28, PCB-52, PCB-101, PCB-138, PCB 153 and PCB-180); the commodities were divided into 48 basic groups. The exposure dose values based on actual and model consumption were calculated for an average inhabitant as well as for children of various age groups, and compared with the Tolerable Daily Intake value (TDI; 0.4 microg kg(-1) body weight per day). Knowledge and experience from a variety of projects running since 1991, as well as the database of food contaminants of the Centre for Evaluation of Contaminant Occurrence established at the Food Research Institute were utilized. The results of the evaluation suggested, that during the period of observation, the exposure of the Slovak Republic population to PCBs was relatively low. Taking actual consumption and mean findings as the basis, the daily exposure doses of PCBs ranged between 3.1% and 6.5% TDI, the corresponding figures for median value and the 95th percentile being between 1.4% and 4.0%, and between 7.5% and 20.0%, respectively. PMID- 17412475 TI - An examination of consumer exposure to caffeine from retail coffee outlets. AB - OBJECTIVE: To analyse the distribution of caffeine doses obtainable from espresso coffee sold by a sample of commercial coffee vendors located on the Gold Coast, Qld, Australia. DESIGN: A cross section of "Espresso/short black" coffee samples were purchased and analysed for their caffeine content using micellar electrokinetic capillary chromatography (MEKC). Coffees were collected using systematic cluster sampling across five major retail centres. RESULTS: Ninety seven espresso samples were analysed. The mean (+/-SD) quantity of caffeine was 106+/-38 mg/serve with a concentration of 2473+/-1092 mg/l. There was considerable variation in caffeine content. The range per serve was 25-214 mg whilst the concentration range was 580-7000 mg/l. Twenty-four samples (24.7%) contained 120 mg of caffeine or higher and 12 samples (12.3%) exceeded 167 mg per serve. CONCLUSIONS AND IMPLICATIONS: The number of heavily caffeinated samples differentiates these findings from frequently cited caffeine values and supports similar data recently collected throughout the United Kingdom. As a result, the accuracy of any previous intake modelling regarding caffeine use in the Australian population is in doubt. The present data suggests that the probability of consumer exposure to high caffeine doses is greater than previously anticipated. Greater sample numbers from a broader selection of venues is required to confirm the extent of caffeine content variance within retail ground coffees. PMID- 17412476 TI - [Uni-axial tissular extension for covering skin defects of the limbs. A 31 cases review]. AB - INTRODUCTION: The skin property to adapt to external constraints is widely used in plastic surgery. Tissue expansion is the most known and codified application. Tissue extension appears to be an attractive alternative. The authors propose the usage of a simplified procedure to deal with skin loss in the superior or inferior members. MATERIAL AND METHOD: Thirty-one patients underwent uni-axial traction between February 2000 and October 2003. RESULTS: Closure of skin loss on the upper member has been obtained in 6 days and in 8 days on the inferior member. DISCUSSION: Efficiency, reliability and no subsequent aftermaths are strong arguments in favor of the development of a simplified extension procedure. CONCLUSION: The tissue extension procedure, although scarcely used is an attractive procedure for the coverage of skin losses. PMID- 17412477 TI - [About cover of ballistic wounds of distal third of leg in civilian ways]. AB - Major causes of ballistic wounds are shooting accidents. Goals of this work are to suggest a position about ballistic wounds of leg's distal third. Reconstruction of ballistic wounds of leg's distal third is difficult, because of lack of laxity and veinous drainage in distal third; and characteristics of ballistic wounds. Ballistic traumas give a temporary cavity, which induce levelled wounds of neurovascular pedicules. We present three cases. One patient benefits from a sural flap, whereas the two others benefit respectively from a free parascapular flap and a free latissimus dorsi flap with free osseous graft. Functional results were considered as good. Treatment of ballistic wounds ask for certain specificities: 1) meticulous evaluation of wounds; 2) evaluation of vascular axis of leg. Debridments are numerous under general anaesthesia before a reconstruction with axial regional flap or free flap, which vessels are anastomosed distant. Reconstruction must be done before the tenth day. PMID- 17412478 TI - Importance of public information and perception for managing recreational activities in the Peel-Harvey estuary, Western Australia. AB - Surveys of water recreational activities were conducted in the Peel-Harvey estuary. Channels used by recreationists to gain information about water quality, the perception of water quality and resulting behaviour were investigated. This study showed that personal perception, local knowledge and history, absence of warnings and residency were major factors contributing to risk perception and behaviour in this recreational community. Management strategies should take this information into account to achieve maximal outcomes. PMID- 17412479 TI - Sediment concentration and turbidity changes during culvert removals. AB - The concentrations of sediment and turbidity in stream water were monitored during culvert removals to determine the short term effects of road obliteration. Sediment concentration was measured at 11 stream crossings among two locations in Idaho and one in Washington. Sediment concentration immediately below the culvert outlet exceeded levels above the culvert outlet by at least three orders of magnitude at all stream crossings. Sediment yields ranged from 170 to less than 1kg in the 24-h period following culvert removal. Turbidity exceeded the regulatory limits during culvert removal at all locations monitored in this study and remained above the limits beyond the monitoring periods of 24h at four of the locations. Sediment concentrations 100m downstream of the culvert outlet were reduced by an order of magnitude, but did not change the turbidity values sufficiently to meet regulatory limits. Sediment concentrations an average of 810m downstream of the culvert outlet were similar to sediment concentrations above the culvert for the entire excavation period and turbidity regulations were met. Mitigation consisting of two straw bales placed in the stream caused a significant reduction in sediment yield from an average of 67kg to an average of 1.6kg. PMID- 17412480 TI - Coupling a 3D patch model and a rockfall module to assess rockfall protection in mountain forests. AB - Many forests in the Alps are acknowledged for protecting objects, such as (rail) roads, against rockfall. However, there is a lack of knowledge on efficient silvicultural strategies and interventions to maintain these forests at optimal protection level. Therefore, assessment tools are required that quantify the rockfall protection effect of forest stands over time, and thereby provide the ability to evaluate the necessity and effect of management interventions. This paper introduces such a tool that consists of a 3D rockfall module embedded in the patch based forest simulator PICUS. The latter is extended for this study with a new regeneration module. In a series of experiments the new combined simulation tool is evaluated with regard to parameter sensitivity, model intercomparison experiments with recently proposed algorithms from the literature, and the ability to respond realistically to different management regimes in rockfall protection forests. Results confirm the potential of the new tool for realistic simulation of rockfall activity in heterogeneous mountain forests, but point at the urgent need to improve the knowledge base on the interaction of understory and rockfall activity. Further work will focus on model validation against empirical rockfall data, and include reduced tree vitality due to damage from boulder collisions as well as the explicit consideration of downed dead wood. PMID- 17412481 TI - Ecological scenarios analyzed and evaluated by a shallow lake model. AB - We applied the complex ecosystem model EMMO, which was adopted to the shallow lake Muggelsee (Germany), in order to evaluate a large set of ecological scenarios. By means of EMMO, 33 scenarios and 17 indicators were defined to characterize their effects on the lake ecosystem. The indicators were based on model outputs of EMMO and can be separated into biological indicators, such as chlorophyll-a and cyanobacteria, and hydro-chemical indicators, such as phosphorus. The question to be solved was, what is the ranking of the scenarios based on their characterization by these 17 indicators? And how can we handle high quantities of complex data within evaluation procedures? The scenario evaluation was performed by partial order theory which, however, did not provide a clear result. By subsequently applying the hierarchical cluster analysis (complete linkage) it was possible to reduce the data matrix to indicator and scenario representatives. Even though this step implies losses of information, it simplifies the application of partial order theory and the post processing by METEOR. METEOR is derived from partial order theory and allows the stepwise aggregation of indicators, which subsequently leads to a distinct and clear decision. In the final evaluation result the best scenario was the one which defines a minimum nutrient input and no phosphorus release from the sediment while the worst scenario is characterized by a maximum nutrient input and extensive phosphorus release from the sediment. The reasonable and comprehensive results show that the combination of partial order, cluster analysis and METEOR can handle big amounts of data in a very clear and transparent way, and therefore is ideal in the context of complex ecosystem models, like that we applied. PMID- 17412482 TI - Quantitative ultrasound measurements in diabetic and nondiabetic patients with end-stage renal disease. AB - The aim of the study was to assess skeletal status in diabetic and nondiabetic subjects with end-stage renal disease (ESRD). One hundred twenty-three patients with ESRD (57 patients with diabetes: 9 type 1 and 48 type 2) and 66 nondiabetic patients were evaluated. Control group comprised 1541 subjects (614 males and 927 females). Diabetes and/or renal insufficiency was the only reason of bone disease and, in control group, no factors known to influence bone metabolism (chronic diseases or prolonged medications) were noted. Skeletal status was evaluated by quantitative ultrasound measurements at the hand phalanges using DBM 1200 (IGEA, Carpi, Italy), which measures amplitude-dependent speed of sound (Ad-SoS [m/s]). Because of some differences in mean age in subgroups of patients and controls, comparisons were performed using values of Z-score. In all diabetic patients, Z score was significantly higher compared with nondiabetics (p < 0.05). In all type 1 diabetes patients, Z-score was significantly lower than in all nondiabetic patients (p < 0.05) and in patients with type 2 diabetes (p < 0.001). Z-score was also significantly lower in type 2 diabetics than in nondiabetic females (p < 0.00001) but did not differ in males. Comparisons between Z-scores in controls and patients showed that Z-score in nondiabetic females was significantly lower than in female controls (p < 0.000001), and in nondiabetic males--diabetic type 2 males as well as females--Z-score did not differ vs. results in adequate control group. Z-score was significantly lower in patients with diabetes type 1 vs. all controls (p < 0.001). Correlation analysis showed in all nondiabetic patients that Z-score was negatively affected by duration time of dialysis (r = -0.37, p < 0.01) and parathyroid hormone (PTH) serum level (r = -0.35, p < 0.01). In patients with type 1 diabetes, only PTH influenced significantly Z-score (r = 0.76, p < 0.05) and, in patients with type 2 diabetes, no significant correlations were obtained. Subjects with type 1 diabetes seemed to be sensitive for skeletal disturbances in a course of renal insufficiency, whereas subjects with type 2 diabetes did not show such skeletal pathology as shown by ultrasound measurements at hand phalanges. PMID- 17412483 TI - Ultrasonographic endometrial features in tubal pregnancy: are they predictive factors of successful medical treatment? AB - The aim of this study was to explain possible relationships in cases of ectopic pregnancy between sonographic endometrial features and treatment outcome following methotrexate (MTX) injection. A total of 157 cases of unruptured tubal pregnancy were diagnosed and treated systemically with MTX. The sonographic endometrial pattern, endometrial thickness and treatment outcome were retrospectively analyzed. There were four types of endometrial patterns: these were trilaminar, homogeneous, heterogeneous and pseudosac. Success rates following MTX treatment were different for each group: these were 64.7%, 78.4%, 50.0% and 46.2%, respectively (p = 0.0129). And the risk of treatment failure was increased 3.64 times (95% CI 1.56 to 8.49) in heterogeneous thick pattern and 4.24 times (95% CI 1.18 to 15.23) in pseudosac pattern. In conclusion, when tubal pregnancy was seen in a pseudogestational sac pattern or a heterogeneous pattern, the failure rate of medical treatment was increased. This can serve as a reference for considering the decision about medical treatment in women with unruptured tubal pregnancy. PMID- 17412484 TI - Constant delay in adapted cerebral response to light stimulation in premature neonates: a transcranial Doppler study. AB - Transcranial Doppler has been used previously to determine hemodynamic response to sensorial stimulation in adults but not in the neonatal period. Cerebral blood flow velocity (CBFV) was recorded in both middle cerebral arteries (MCA) and posterior cerebral arteries (PCA) in 100 premature neonates (male 56; female 44; 26 to 36 weeks gestation) from birth to 12 months. Brain development was evaluated on clinical examination. No difference in velocities was noted on prestimulation recordings from birth to 12 months. During light stimulation before six months, the CBFV was reduced (-11 +/- 6% in MCA and -13 +/- 5% in PCA compared with baseline values). After stimulation, the CBFV was increased (+6.7 +/- 3% in MCA and + 10.5 +/- 4% in PCA compared with baseline values). This study is in favor of cortex maturation in normal premature neonates at only six months. Consequently, functional transcranial Doppler technique will be helpful for the diagnosis of abnormal maturation timing in neonates with possible developmental retardation. PMID- 17412485 TI - Velocity measurements using a single transmitted linear frequency-modulated chirp. AB - Velocity measurement is a challenge for a variety of remote sensing systems such as ultrasonic and radar scanners. However, current Doppler-based techniques require a comparatively long data acquisition time. It has been suggested to use coded signals, such as linear frequency-modulated signals (chirp), for ultrasonic velocity estimation by extracting the needed information from a set of several sequential coded pulses. In this study, a method for velocity estimation using a single linear frequency-modulated chirp transmission is presented and implemented for ultrasonic measurements. The complex cross-correlation function between the transmitted and reflected signals is initially calculated. The velocity is then calculated from the phase of the peak of the envelope of this cross-correlation function. The suggested method was verified using computer simulations and experimental measurements in an ultrasonic system. Applying linear regression to the data has yielded very good correlation (r = 0.989). With the suggested technique, higher frame rates of velocity mapping can be potentially achieved relative to current techniques. Also, the same data can be utilized for both velocity mapping and image reconstruction. PMID- 17412486 TI - Destruction thresholds of echogenic liposomes with clinical diagnostic ultrasound. AB - Echogenic liposomes (ELIP) are submicron-sized phospholipid vesicles that contain both gas and fluid. With antibody conjugation and drug incorporation, these liposomes can be used as novel targeted diagnostic and therapeutic ultrasound contrast agents. The utility of liposomes for contrast depends upon their stability in an acoustic field, whereas the use of liposomes for drug delivery requires the liberation of encapsulated gas and drug payload at the desired treatment site. The objective of this study was twofold: (1) to characterize the stability of liposome echogenicity after reconstitution and (2) to quantitate the acoustic destruction thresholds of liposomes as a function of peak rarefactional pressure (P(r)), pulse duration (PD) and pulse repetition frequency (PRF). The liposomes were insonified in an anechoic sample chamber using a Philips HDI 5000 diagnostic ultrasound scanner with a L12-5 linear array. Liposome stability was evaluated with 6.9-MHz fundamental and 4.5-MHz harmonic B-mode pulses at various P(r) at a fixed PRF. Liposome destruction thresholds were determined using 6.0 MHz Doppler pulses, by varying the PD with a fixed PRF of 1.25 kHz and by varying the PRF with a fixed PD of 3.33 micros. Videos or freeze-captured images were acquired during each insonation experiment and analyzed for echogenicity in a fixed region of interest as a function of time. An initial increase in echogenicity was observed for fundamental and harmonic B-mode imaging pulses. The threshold for acoustically driven diffusion of gas out of the liposomes using 6.0 MHz Doppler pulses was weakly dependent upon PRF and PD. The rapid fragmentation thresholds, however, were highly dependent upon PRF and PD. The quantification of acoustic destruction thresholds of ELIP is an important first step in their development as diagnostic and drug delivery agents. PMID- 17412487 TI - Editorial Comment on: association between physical activity and urinary incontinence in a community-based elderly population aged 70 years and over. PMID- 17412488 TI - Association between physical activity and urinary incontinence in a community based elderly population aged 70 years and over. AB - OBJECTIVES: The objective of the present study was to evaluate the association between physical activity (PA) levels and urinary incontinence (UI) in a community-based elderly population aged > or =70 yr. METHODS: This population based cross-sectional survey was conducted in 2003 using an extensive health interview for each participant. A self-reported single-item questionnaire was used to estimate different levels of PA in each subject. The prevalence of UI was estimated by the self-administered International Consultation on Incontinence Questionnaire. The study population included 676 Japanese men and women. RESULTS: The prevalence of UI was 25% (34% in women and 16% in men). After adjustment for potential confounding factors, the odds ratio (95% confidence interval) of UI compared with the lowest PA group was 0.71 (0.47-1.09) and 0.58 (0.35-0.96) in subjects exhibiting middle and high levels of PA, respectively (p for trend = 0.02). CONCLUSIONS: High PA level was independently related to a lower self reported prevalence of UI in a community-dwelling elderly population aged > or =70 yr. Although this cross-sectional study cannot demonstrate a temporal relationship between PA and the onset of UI, the findings suggest that PA may have a potentially beneficial effect on the prevention of UI. A prospective study or randomized trials are required to clarify the causality. PMID- 17412489 TI - Extended 21-sample needle biopsy protocol for diagnosis of prostate cancer in 1000 consecutive patients. AB - OBJECTIVE: To prospectively evaluate the diagnostic yield of a 21-sample ultrasound-guided needle biopsy protocol as the initial diagnostic strategy for detection of prostate cancer. MATERIALS AND METHODS: Between December 2001 and October 2005, 1000 consecutive patients underwent 21-sample needle biopsies under local anesthesia, comprising sextant biopsies, 3 additional posterolateral biopsies in each peripheral zone, 3 biopsies in each transition zone (TZ), and 3 biopsies in the midline peripheral zone. Each prostate core was numbered and analyzed separately. The patients were divided into subgroups according to the result of digital rectal examination (DRE), serum prostate-specific antigen (PSA), and prostate volume. We evaluated the cancer detection rate overall and in each subgroup. We compared the results of our biopsy protocol to those from 6-, 12-, and 18-core biopsy protocols by analyzing only those cores from our protocol that would correspond to these biopsy schemes. RESULTS: Cancer detection rates using 6 biopsy samples (sextant biopsies only), 12 samples (sextant plus lateral biopsies), 18 samples (sextant, lateral, and TZ biopsies), and 21 samples (sextant, lateral, TZ, plus midline biopsies) were 31.7%, 38.7%, 41.5%, and 42.5%, respectively. The 12-sample procedure improved the cancer detection rate by 22% compared with the 6-sample procedure (p=0.0001). The improvement in the diagnostic yield was most marked in patients with a prostate volume > or =55 ml (36.9%), in patients with normal DRE (26.6%), and in patients with PSA<4 (37.5%). The addition of TZ biopsies to a 12-biopsy scheme increased the diagnostic yield by 7.2% overall (p=0.023). Only 10 of 425 (2.3%) patients were diagnosed on the sole basis of midline biopsies. CONCLUSIONS: Patients with suspected localized prostate cancer should be offered at least 12 biopsies in the peripheral zone and far lateral peripheral zone (statistically significant). TZ biopsies have to be considered, because these biopsies improve the diagnostic yield. For patients with abnormal DRE and/or PSA> or =20 ng/ml, the 6-biopsy scheme seems sufficient (statistically), but 6 far lateral peripheral zone biopsies as well as the TZ biopsies add little incremental value (not significant). Evidence does not support the use of routine midline peripheral zone needle biopsies in the initial biopsy to enhance the detection of prostate cancer. PMID- 17412490 TI - Preoperative serum testosterone level as an independent predictor of treatment failure following radical prostatectomy. AB - OBJECTIVES: Preoperative low serum testosterone (TS) level has been reported to be associated with adverse pathologic results in patients with clinically localized prostate cancer (pCA) treated with radical prostatectomy (RP). However, prior studies failed to show prognostic impact of preoperative low TS in these patients. The aim of this study was to investigate the relationship between preoperative TS and prostate-specific antigen (PSA) failure in these patients. METHODS: Of 304 patients diagnosed with clinically localized pCA who had been treated with RP alone, 272 patients whose preoperative TS level had been measured were eligible for this analysis. Postoperative TS levels were also available in 222 of the 272 patients. Cox proportional hazard model was used to elucidate factors predictive for PSA failure. RESULTS: Of the 272 patients 49 had low (< 300 ng/dl) and 223 had normal preoperative TS level. In a stepwise multivariate analysis, preoperative TS (p = 0.021) was an independent and significant predictor of PSA failure along with RP Gleason score (p = 0.006), surgical margin status (p = 0.0001), and PSA (p = 0.0001). Five-year PSA failure-free survival rate of the patients with preoperative low TS (67.8%) was significantly worse than that with normal TS (84.9%) (p=0.035). Serum TS levels increased significantly after RP (p < 0.0001). The increment of TS level in preoperative low TS group was significantly greater than that in preoperative normal TS group (p = 0.0003). CONCLUSIONS: The current results demonstrated that preoperative TS level is an independent and significant predictor of PSA failure after RP in patients with clinically localized pCA. PMID- 17412491 TI - Selective anterograde amnesia with thalamus and hippocampal lesions in neuro Behcet's disease. AB - Anterograde amnesia and minimal retrograde amnesia with thalamic and hippocampal lesions in neuro-Behcet's disease is rare. A 50-year-old man presented with forgetfulness and severe memory disturbance after suffering multiple oral and genital aphthous ulcers with erythema nodosum. A neurological examination and a neuropsychological assessment revealed prominent anterograde memory impairment without focal neurological deficits. On brain MRI there were high signal intensity lesions involving right anterior thalamus, left posterior basal ganglia, and left hippocampus. This is a quite selective anterogrde memory deficit in a case of neuro-Behcet's disease caused by parenchymal lesions in the thalamus and hippocampus. PMID- 17412492 TI - Treatment of an iatrogenic vertebral artery laceration with the Symbiot self expandable covered stent. AB - Vertebral artery laceration is difficult to treat surgically. Endovascular treatment with balloon expandable covered stents often fails due to their rigidity and poor navigability. We present a case of iatrogenic vertebral artery laceration where endovascular treatment with a balloon expandable covered stent failed. Eventually a self expandable symbiot covered stent was deployed over the laceration, securing hemostasis and preserving the vessel patency. The newer self expandable covered stents seem promising in the treatment of vertebral artery injury. PMID- 17412493 TI - 1alpha,25-Dihydroxyvitamin D(3) antiproliferative actions involve vitamin D receptor-mediated activation of MAPK pathways and AP-1/p21(waf1) upregulation in human osteosarcoma. AB - The molecular mechanisms underlying antiproliferative actions of the steroid 1alpha,25-dihydroxy vitamin D(3) (1,25D) in human osteosarcoma cells are known only partially. To better understand the signaling involved in 1,25D anti tumorigenic properties in bone, we stably silenced vitamin D receptor (VDR) expression in the human osteosarcoma SaOS-2 cell line. We found that 1,25D treatment reduced cell proliferation by approximately 25% after 3 days only in SaOS-2 cells expressing native levels of VDR protein, and involved activation of MAPK/AP-1/p21(waf1) pathways. Both sustained (3 days) and transient (15min) 1,25D treatment activated JNK and ERK1/2 MAPK signaling in a nongenomic VDR-dependent manner. However, only sustained exposure to hormone led to upregulation of p21 and subsequent genomic control of the cell cycle. Specific blockade of MEK1/MEK2 cascade upstream from ERK1/2 abrogated 1,25D activation of AP-1 and p21, and subsequent antiproliferative effects, even in the presence of a nuclear VDR. We conclude that 1,25D-induced inhibition of human osteosarcoma cell proliferation occurs via sustained activation of JNK and MEK1/MEK2 pathways downstream of nongenomic VDR signaling that leads to upregulation of a c-Jun/c-Fos (AP-1) complex, which in turn modulates p21(waf1) gene expression. Our results demonstrate a cross-talk between 1,25D/VDR nongenomic and genomic signaling at the level of MAP kinase activation that leads to reduction of cell proliferation in human osteosarcoma cells. PMID- 17412494 TI - Electrokinetic removal of 2,6-dichlorophenol and diuron from kaolinite and humic acid-clay system. AB - This paper presents the results of a study on the electrokinetic treatment of kaolinite and humic acid kaolinite complexes spiked with 2,6-dichlorophenol or 3 (3,4-dichlorophenyl)-1,1-dimethylurea (diuron). In particular, the attention was paid to the interaction between solid surface and dissolved organics: the effects of contaminant sorption as well as the physicochemical reactions on the efficiency of electrokinetic remediation were investigated. Using a 3V/cm voltage gradient, approximately 90% of diuron was removed from kaolinite after one water pore volume was collected in the cathode reservoir, but much lower efficiency was obtained in the electrokinetic removal of this compound from humic acid-coated kaolinite. The results also showed that partial degradation of the contaminant occurred during electrokinetic treatment of kaolinite clay spiked with 2,6 dichlorophenol: the contamination in the clay could be remediated by the combination of electrokinetic extraction and electrochemical reactions. PMID- 17412495 TI - Ultrasonic reduction of excess sludge from the activated sludge system. AB - Sludge treatment has long become the most challenging problem in wastewater treatment plants. Previous studies showed that ozone or chlorine effectively liquefies sludge into substrates for bio-degradation in the aeration tank, and thus reduces the excess sludge. This paper employs ultrasound to reduce the excess sludge from the sequential batch reactor (SBR) system. Partial sludge was disintegrated into dissolved substrates by ultrasound in an external sono-tank and was then returned to the SBR for bio-degradation. The results showed that ultrasound (25kHz) effectively liquefied the sludge. The most effective conditions for sludge reduction were as following: sludge sonication ratio of 3/14, ultrasound intensity of 120kW/kgDS, and sonication duration of 15min. The amount of excess sludge was reduced by 91.1% to 17.8mg/(Ld); the organic content and settleability of sludge in the SBR were not impacted. The chemical oxygen demand (COD) removal efficiency was 81.1%, the total nitrogen (TN) removal efficiency was 17-66%, and high phosphorus concentration in the effluent was observed. PMID- 17412496 TI - Effect of filtration velocity and dust concentration on cake formation and filter operation in a pilot scale jet pulsed bag filter. AB - Bag filters are used for the removal of fine solid particles from process gases. Thus, understanding the filter cake build up and its properties is a subject of interest. The filter cakes properties may depend on many factors like, for example, filtration velocity and dust concentration. The effect of dust concentration and filtration velocity on filtration time, specific cake resistance and mean cake density is investigated in a pilot scale jet pulsed bag filter. An in situ optical system is used to measure cake thickness distributions on the filter surface. Additionally, the operation is simulated using a one dimensional model and results are compared with experiments. The experimental results indicate that cake density and specific resistance increase with increasing velocity at constant dust concentration. The effect of dust concentration on filter cake density and specific resistance is small. PMID- 17412497 TI - Studies on the applicability of alginate-entrapped Chryseomonas luteola TEM 05 for heavy metal biosorption. AB - Chryseomonas luteola TEM 05 cells were entrapped both in alginate and chitosan coated alginate beads. Biosorption of metal ions on alginate beads was investigated by using a batch stirred system at pH 6.0, 25 degrees C, in initial metal concentration of 1.92 mM of Cr6+, 0.89 mM Cd2+ and 1.69 mM Co2+. Then, a process of competitive biosorption of these metal ions was described and compared to single metal ion adsorption in solution. The apparent equilibrium biosorption was reached within the 180 min of contact for all metals. Although the competitive biosorption capacities of the beads for all metal ions were lower than those of single conditions, Cd2+ biosorption on alginate and alginate chitosan beads did not change significantly. PMID- 17412498 TI - On-line system for preconcentration and determination of metals in vegetables by inductively coupled plasma optical emission spectrometry. AB - A procedure has been developed for the simultaneous determination of trace amounts of cadmium, copper, chromium, nickel and lead in digested vegetable samples. The method involves solid-phase extraction of the metals using a minicolumn of Amberlite XAD-4 modified with dihydroxybenzoic acid (DHB) and detection by Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES). The elution of the metals from minicolumn was performed with 1.0 molL(-1) hydrochloric acid. Variables associated with flow preconcentration system performance, such as pH, buffer concentration, eluent concentration and sampling flow rate, were optimized. The developed procedure provides enrichment factors of 100, 72, 16, 91 and 53, for cadmium, copper, chromium, nickel and lead, respectively. Detection limits (3sigma(B)) were 0.02 (Cd), 0.23 (Cu), 0.58 (Cr), 0.060 (Ni) and 0.54 (Pb)microgL(-1). The procedure was applied for determination of metals in samples of guarana and cabbage. The accuracy of the method was checked by the analysis of a certified reference material (NIST 1571, Orchard leaves). Results found were in agreement with certified values. PMID- 17412499 TI - Semicontinuous microcosm study of aerobic cometabolism of trichloroethylene using toluene. AB - A semicontinuous slurry-microcosm method was applied to mimic trichloroethylene (TCE) cometabolic biodegradation field results at the Que-Jen in-situ pilot study. The microcosm study confirmed the process of aerobic cometabolism of TCE using toluene as the primary substrate. Based on the nucleotide sequence of 16S rRNA genes, the toluene-oxidizing bacteria in microcosms were identified, i.e. Ralstonia sp. P-10 and Pseudomonasputida. The first-order constant of TCE degradation rate was 0.5 day(-1) for both Ralstonia sp. P-10 and P.putida. The TCE cometabolic-biodegradation efficiency measured from the slurry microcosms was 46%, which appeared pessimistic compared to over 90% observed from the in-situ pilot study. The difference in the TCE cometabolic-biodegradation efficiency was likely due to the reactor configurations and the effective time duration of toluene presence in laboratory microcosms (1 days) versus in-situ pilot study (3 days). The results of microcosm experiments using different toluene-injection schedules supported the hypothesis. With a given amount of toluene injection, it is recommended to maximize the effective time duration of toluene presence in reactor design for TCE cometabolic degradation. PMID- 17412500 TI - Effects of local anesthetics on opioid inhibition of calcium current in rat dorsal root ganglion neurons. AB - Neuraxial analgesia is often provided using a mixture of local anesthetics and opioids. This combination of agents provides better pain relief and is generally associated with fewer side effects than when either drug is given alone. Local anesthetics have been shown to alter signaling of other G protein-coupled receptors, but little is known about their effect on opioid receptor signaling. Because opioids produce analgesia at least in part by inhibiting presynaptic Ca channels, we have evaluated the effects of tetracaine and bupivacaine on opioid mediated inhibition of Ca channels in dorsal root ganglion neurons. The mu-opioid specific agonist DAMGO (1microM) inhibited Ca channels in both the absence and presence of tetracaine (50 or 100muM). However, the extent of DAMGO inhibition in the presence of both concentrations of tetracaine was less than that observed in the absence of tetracaine. DAMGO inhibition decreased from 39.2+/-24.4% in control to 34.2+/-24.4% with 50microM tetracaine (n=16; p<0.05), and from 40.5+/ 19.6% in control to 34.6+/-20.5% with 100microM tetracaine (n=10; p<0.05). Similar results were seen with bupivacaine. Tetracaine also decreased the voltage dependent facilitation of Ca channel currents when G proteins were activated by either DAMGO or the non-hydrolyzable GTP analogue (GTPgammaS), suggesting that tetracaine weakens the interaction between G protein betagamma subunits and the Ca channel. Overall, these results suggest that local anesthetics decrease opioid inhibition of Ca channel activity by interfering with the GTP-mediated signal transduction between opioid receptors and Ca channels. PMID- 17412501 TI - Bone marrow stromal cells of transgenic mice can improve the cognitive ability of an Alzheimer's disease rat model. AB - This study investigated the effects of bone marrow stromal cells transplantation on Alzheimer's disease (AD). Bone marrow stromal cells (BMSC) were obtained from the bone marrow of transgenic mice expressing green fluorescent protein and transplanted into the hippocampus of rats, which had received an injection of beta amyloid protein into the hippocampus 8 days earlier. Morris Water Maze test was used to observe behavior 2 weeks after transplantation. The survival and differentiation of the grafts were studied immunohistochemically. Behavior improved significantly in the transplanted group. The transplanted BMSC survived and presented ChAT-like neurons, indicating that these transplanted cells might differentiate into cholinergic neurons and the procedure could be a promising therapy for Alzheimer's disease. PMID- 17412502 TI - Anaesthetic binding sites for etomidate and propofol on a GABAA receptor model. AB - Investigating the molecular basis of general anaesthetic activity at the GABA(A) ligand-gated ion channel is challenging due to the wide structural diversity among known general anaesthetics, and the lack of an experimental structure for the GABA(A) protein. In this molecular modelling study, two distinct binding cavities were identified within the beta(2) subunit of the transmembrane domain in a molecular model of the GABA(A) protein. The first, located near the centre of the alpha-helical bundle, contains Asn265 (TM2), which is essential for modulation by etomidate. The second, located near the TM1, TM3 and TM4 segments close to the membrane-extracellular interface, is capped by Met286 (TM3), a residue thought to be involved in the propofol binding site. Potential interactions of etomidate and propofol with other side-chains were also identified. PMID- 17412503 TI - Glutamatergic vestibular neurons express Fos after vestibular stimulation and project to the NTS and the PBN in rats. AB - In this study, retrograde tracing method combined with phosphate-activated glutaminase (PAG) and Fos immunofluorescence histochemistry was used to identify glutamatergic vestibular nucleus (VN) neurons receiving vestibular inputs and projecting to the nucleus of the solitary tract (NTS) and the parabrachial nucleus (PBN). Conscious animals were subjected to 120 min Ferris-wheel like rotation stimulation. Neuronal activation was assessed by Fos expression in the nucleus of VN neurons. After Fluoro-gold (FG) injection into the caudal NTS, approximately 48% FG-labeled VN neurons were immunoreactive for PAG, and about 14% PAG/FG double-labeled neurons co-existed with Fos. Following FG injection into the PBN, approximately 56% FG-labeled VN neurons were double-labeled with PAG, and about 12% of the PAG/FG double-labeled neurons also expressed Fos. Careful examination of the typology and distribution pattern of these PAG immunoreactive neurons indicated that the vast majority of these neurons were glutamatergic rather than GABAergic. These results suggest that PAG immunoreactive VN neurons might constitute excitatory glutamatergic VN-NTS and VN PBN transmission pathways and these pathways might be involved in vestibulo autonomic reflexes during vestibular stimulation. PMID- 17412504 TI - Age-dependent changes in dopaminergic projections from the substantia nigra pars compacta to the neostriatum. AB - Age-dependent changes in dopaminergic (DA) innervation of the neostriatum (Str) were studied in male F344/N rats. Projections from the substantia nigra pars compacta (SNc) to the neostriatum were quantified using electrophysiological methods at age points from 6 to 24 months. The percentage of DA neurons activated antidromically by electrical stimulation (P-index) of Str increased between 18 and 24 months. Additionally, the percentage of DA neurons showing multiple antidromic latencies from striatal stimulation (M-index), which suggests axonal branching of individual DA neurons, increased significantly between 6 and 12 months and 6 and 24 months. These results suggest that DA neurons exhibit increased axonal branching in the aged brain. PMID- 17412505 TI - Specific and artifactual labeling in the rat spinal cord and medulla after injection of monosynaptic retrograde tracers into the diaphragm. AB - The use of fluorescent dyes has been a major improvement for paths tracing studies. However, these tracers present different properties and have to be chosen carefully. The present study compares the ability of different tracers to specifically label phrenic motoneurons (PMNs) innervating the rat diaphragm. The administration of fluorogold (FG) from the transected phrenic nerve specifically labeled PMNs in the ipsilateral spinal cord. However, when FG was injected into one hemidiaphragm, in addition with ipsilateral PMNs, a less intense artifactual labeling was observed in the spinal cord (mainly in contralateral PMNs) and in the medulla oblongata (mainly in the area postrema and cranial motor nuclei). Similar results were observed using horseradish peroxidase, while no labeling was observed after injection of nuclear yellow or diamidino yellow into the diaphragm. By contrast, the dextran amine fluororuby (FR) and the carbocyanine DiAsp selectively and exclusively labeled ipsilateral PMNs 2 or 3 weeks after injection into the diaphragm, respectively. The lipophilic properties of DiAsp and the high molecular weight of FR may prevent their diffusion to adjacent tissues and into the blood stream which seems to account for the artifactual labeling observed with the other tracers. The higher homogeneity and quality of the labeling observed with FR compared to DiAsp make it the most appropriate tracer for the specific monosynaptic fluorescent labeling of PMNs after injection into the diaphragm. PMID- 17412506 TI - A mutation screening by DHPLC of PSEN1 and APP genes reveals no significant variation associated with the sporadic late-onset form of Alzheimer's disease. AB - Alzheimer's disease (AD), the most common cause of dementia in the elderly, is usually divided into familial and sporadic forms, according to family history. The familial form has often been reportedly caused by mutations in amyloid precursor protein (APP), presenilin-1 (PSEN1), or presenilin-2 (PSEN2) genes, whereas the genetic component for the sporadic form is less clear. We carried out mutation screening in exons 16 and 17 of APP, and in exons 3, 4, 5, 6, 7, 10 of PSEN1 genes in patients with the sporadic late-onset form of AD (LOAD). The aim of this study was to ascertain whether any variation in these genes, besides that of the well-known apolipoprotein E common polymorphism, could be involved in the onset of the disease. To search for the single nucleotide substitutions, we examined 172 LOAD patients by the denaturing high-performance liquid chromatography (DHPLC) technique. Only one same-sense mutation in exon 4 of PSEN1 gene (N32) was observed in this patient group. We concluded that the variation in the screened exons of the APP and PSEN1 genes, reportedly associated with familial AD, is not present in LOAD. PMID- 17412507 TI - Microarray expression analysis of the early N-methy-N-nitrosourea-induced retinal degeneration in rat. AB - The study was undertaken to investigate the gene expressions in N-methy-N nitrosourea (MNU)-induced rat retinal degeneration (RD) by performing microarray analysis of retinal RNA at 12h. All rats were randomly divided into a normal group, a 12h model group and a 24h model group. Rats in the two model groups received a single intraperitoneal injection of 40 mg/kg body weight of MNU, while those in the normal group were injected with equivalent volume of physiological saline. After 12h and 24h of the injection, rats in each respective group were sacrificed, respectively. One eye of each animal was used for hematoxylin and erosin (H&E) staining, and fresh retinas of the other eye of each animal in the both normal group and 12h model group were used to extract total RNA, which was analyzed by microarray and real time RT-PCR. Retinal histological alteration was found in the 24h model group. There were 75 genes differently expressed (ratio > or =2.0), including 64 genes up-regulated and 11 genes down-regulated. Seven genes were assayed by real time RT-PCR and demonstrated the same alteration tendency as in microarray analysis. These genes that expressed differently mainly involved signal transduction, development, immune and defense, and apoptosis, etc. The major pathways were MAP-kinase signaling pathways, Toll-like receptor signaling pathway and apoptosis pathway involved. The results suggest that there are significant changes of gene expression in the early stage of MNU-induced RD. These microarray results provide clues to understand the molecular pathways underlying photoreceptor degeneration and indicate directions for future studies. PMID- 17412508 TI - The effect of low-dose insulin on mechanical sensitivity and allodynia in type I diabetes neuropathy. AB - The pathogenesis of diabetic neuropathy is multifactorial, but in general hyperglycemia through polyol and protein glycation pathways is considered to be a key etiological factor. Most likely insulin deficiency, in experimentally induced type I diabetes, contributes to the development of diabetes neuropathy. The aim of this study was to evaluate the in vivo behavioral effect of low-dose insulin on diabetic neuropathy in rats through behavioral testing in hyperglycemic conditions. Mechanical sensitivity and allodynia were tested in streptozotocin (STZ)-induced diabetic rats. After diabetes and neuropathy induction, treatment with low-dose insulin normalized behavioral test results in 37 days, while severe hyperglycemia persisted. Although this study provided no evidence about the role of hypoinsulinemia in the etiology of diabetes neuropathy, the results confirmed that an insulin deficit with impaired insulin signaling and neurotrophic support, rather than hyperglycemia, plays an essential role in the pathophysiology of painful diabetic neuropathy. PMID- 17412509 TI - The neural basis of selection-for-action. AB - The selection of objects in the visual environment is important in everyday life when acting in a goal-directed manner. Here we used functional magnetic resonance imaging (fMRI) to investigate brain activity while healthy subjects (N=15) selectively reached to grasp a three-dimensional (3D) target stimulus presented either in isolation or in the presence of 3D non-target stimuli. A pneumatic MRI compatible apparatus was designed to precisely control the presentation of 3D graspable stimuli within the scanner. During scanning subjects were instructed to reach and grasp towards a target presented at an unknown location either in isolation or flanked by two distractor objects. Results indicated that reaching towards and grasping the target object in the presence of other non-target stimuli was associated with greater activation within the contralateral primary motor cortex and the precuneus as compared to the execution of reach-to-grasp movements towards the target presented in isolation. We conclude that the presence of non-targets evokes a differential level of neural activity within areas responsible for the planning and execution of selective reach-to-grasp movement. PMID- 17412510 TI - Nitric oxide donors attenuate clongenic potential in rat C6 glioma cells treated with alkylating chemotherapeutic agents. AB - 1,3-Bis(2-chloroethyl)-1-nitrosourea (BCNU) kills tumor cells via multiple actions including alkylation and carbamoylation. Previously, we have reported that formation of S-nitrosoglutathione (GSNO) in glioma cells overexpressing inducible nitric oxide synthase (iNOS) contributed to nitric oxide (NO)-dependent carbamoylating chemoresistance against BCNU. To further characterize the effects of NO on alkylating cytotoxicity, colony formation assay was applied to evaluate the effects of various NO donors on rat C6 glioma cells challenged with alkylating agents. We demonstrate that NO donors including GSNO, diethylamine NONOate (DEA/NO), and sodium nitroprusside (SNP) substantially reduced the extent of colony formation in glioma cells treated with alkylating agents, namely methyl methanesulfonate (MMS), N-methyl-N-nitrosourea (MNU), and N-ethyl-N-nitrosourea (ENU). Without alkylating agents these NO-releasing agents alone had no effects on clongenic potential of rat C6 glioma cells. Among these three NO donors used, the effectiveness in potentiating alkylating cytotoxicity is in the order of "GSNO>DEA/NO>SNP" when applied at the same dosages. GSNO also exerted similar synergistic actions reducing the extents of colony formation when co administrated with 1,2-bis(methylsulfonyl)-1-(2-chloroethyl)-hydrazine (compound #1), another alkylating agent that mimics the chloroethylating action of BCNU. Together with our previous findings, we propose that NO donors may be used as adjunct chemotherapy with alkylating agents for such malignant brain tumors as glioblastoma multiforme (GBM). In contrast, production of NO as a result of iNOS induction, such as that occurring after surgical resection of brain tumors, may compromise the efficacy of carbamoylating chemotherapy. PMID- 17412511 TI - Aberrant amino acid transport in fibroblasts from children with autism. AB - Autism is a developmental, cognitive disorder clinically characterized by impaired social interaction, communication and restricted behaviours. The present study was designed to explore whether an abnormality in transport of tyrosine and/or alanine is present in children with autism. Skin biopsies were obtained from 11 children with autism (9 boys and 2 girls) fulfilling the DSM-IV diagnostic criteria for autistic disorder and 11 healthy male control children. Transport of amino acids tyrosine and alanine across the cell membrane of cultured fibroblasts was studied by the cluster tray method. The maximal transport capacity, V(max) and the affinity constant of the amino acid binding sites, K(m), were determined. Significantly increased V(max) for alanine (p=0.014) and increased K(m) for tyrosine (p=0.007) were found in children with autism. The increased transport capacity of alanine across the cell membrane and decreased affinity for transport sites of tyrosine indicates the involvement of two major amino acid transport systems (L- and A-system) in children with autism. This may influence the transport of several other amino acids across the blood brain-barrier. The significance of the findings has to be further explored. PMID- 17412512 TI - Stimulus-evoked opioid inhibition in guinea-pig longitudinal muscle-myenteric plexus strip is modulated by NMDA receptors. AB - Longitudinal muscle-myenteric plexus (LMMP) strips of the guinea-pig ileum were used to investigate the stimulus-evoked endogenous opioid inhibition and its modulation by ionotropic glutamate receptors. Regular cholinergic twitch responses evoked by a short 3-s-field stimulation in intervals of 80s were found reduced after an interposed period of prolonged 40-s-field stimulation. In the presence of a peptidase-inhibitor-cocktail, the cholinergic twitch response following the period of prolonged stimulation was even further reduced as compared to normal Tyrode solution without peptidase inhibitors. In both instances, the impairment of the cholinergic twitch response was completely abolished by naloxone thus demonstrating its opioidergic nature. This endogenous inhibitory opioid effect was significantly mitigated by the NMDA-receptor antagonist MK-801, but not by the AMPA/kainate receptor antagonist CNQX. These results demonstrate by functional experiments that there is a significant opioid mediated inhibition in guinea-pig LMMP preparations evoked by a prolonged electrical stimulation, and that an NMDA antagonist can mitigate the opioid inhibition. PMID- 17412513 TI - Septicemia as a cause of death in burns. PMID- 17412514 TI - Codeine presynaptically inhibits the glutamatergic synaptic transmission in the nucleus tractus solitarius of the guinea pig. AB - Although codeine is the most prominent and centrally acting antitussive agent, the precise sites and mode of its action have not been fully understood yet. In the present study, we examined the effects of codeine on synaptic transmission in second-order neurons of the nucleus tractus solitarius (NTS), which is the first central relay site receiving tussigenic afferent fibers, by using whole-cell patch-clamp recordings in guinea-pig brainstem slices. Codeine (0.3-3 mM) significantly decreased the amplitude of excitatory postsynaptic currents (EPSCs) evoked by electrical stimulation of the tractus solitarius in a naloxone reversible and concentration-dependent manner, but it had no effect on the decay time of evoked EPSCs (eEPSCs). The inhibition of eEPSCs was accompanied by an increased paired-pulse ratio of two consecutive eEPSCs. The inward current induced by application of AMPA remained unchanged after codeine application. A voltage-sensitive K+ channel blocker, 4-aminopyridine (4-AP) attenuated the inhibitory effect of codeine on eEPSCs. These results suggest that codeine inhibits excitatory transmission from the primary afferent fibers to the second order NTS neurons through the opioid receptors that activate the 4-AP sensitive K+ channels located at presynaptic terminals. PMID- 17412515 TI - Differential activation of anterior and midline thalamic nuclei following retrieval of aversively motivated learning tasks. AB - Two thalamic nuclear groups, the anterior thalamic nuclei (ATN) and midline and intralaminar thalamic complex (MITC) have connections to the prefrontal cortex, amygdala, hippocampus and accumbens that are important for learning and memory. However, the anatomical proximity between the ATN and MITC makes it difficult to reveal their roles in memory retrieval of aversive conditioned behavior. To address the issue, we explored the activation of the ATN and MITC, as represented by the expression of the immediate early gene c-fos, following either the retrieval of a conditioned taste aversion (CTA) induced by taste-LiCl pairing (visceral aversion) or of inhibitory avoidance (IA) induced by context-foot shock pairing (somatic aversion) in rats. The anterodorsal (AD) nucleus in the ATN was activated by foot shock and the recall of IA, but not by i.p. injection of LiCl or the recall of CTA. No significant elevation was observed in the other ATN following these treatments. Among nuclei of the MITC, the paraventricular thalamic nucleus (PVT) was activated by the delivery of shock or LiCl and by the recall of both CTA and IA, while the mediodorsal thalamus (MD) and central medial and intermediate thalamus (CM/IMD) were not. The innately aversive taste of quinine did not elevate c-fos expression in either the ATN or MITC. These results suggest that the PVT in the MITC is involved in the processing and retrieval of both taste-malaise and context-shock association tasks, while the AD in the ATN is involved in those of context-shock association only. The difference of the activity between the ATN and MITC demonstrates their functional and anatomical heterogeneity in neural substrates for aversive learning tasks. PMID- 17412517 TI - Unilateral low-frequency stimulation of central piriform cortex inhibits amygdaloid-kindled seizures in Sprague-Dawley rats. AB - The central piriform cortex (cPC) is considered to be critically involved in the generation and propagation of kindled seizures. Our previous study found that low frequency stimulation (LFS) of the cPC inhibits the development process of amygdala kindling. In this study, we determined whether unilateral LFS of the cPC had an inhibitory effect on amygdaloid-kindled seizures in Sprague-Dawley rats. When fully-kindled seizures were achieved by daily amygdala electrical stimulation (2 s train of 1 ms pulses at 60 Hz and 150-300 microA), LFS (15 min train of 0.1 ms pulses at 1 Hz and 50-150 microA) was applied to the ipsilateral or contralateral cPC 1 s after cessation of kindling stimulation for 10 days. LFS of the ipsilateral cPC significantly decreased the incidence of generalized seizures and seizure stage, and shortened cumulative afterdischarge duration and cumulative generalized seizure duration. LFS of the contralateral cPC also significantly decreased the expression of seizure stage, but had no appreciable effect on the generalized seizure incidence, cumulative afterdischarge duration and cumulative generalized seizure duration. On the other hand, LFS of the ipsilateral cPC significantly increased the afterdischarge threshold and further increased the differences of current intensity between afterdischarge threshold and generalized seizure threshold. Our data suggest that LFS of the cPC may be an effective method of inhibiting kindled seizures by preventing both afterdischarge generation and propagation. It provide further evidence that brain regions like the cPC, other than the seizure focus, can serve as targets for deep brain stimulation treatment of epilepsy. PMID- 17412516 TI - Electrophysiological and morphological heterogeneity of slow firing neurons in medial septal/diagonal band complex as revealed by cluster analysis. AB - Slow firing septal neurons modulate hippocampal and neocortical functions. Electrophysiologically, it is unclear whether slow firing neurons belong to a homogeneous neuronal population. To address this issue, whole-cell patch recordings and neuronal reconstructions were performed on rat brain slices containing the medial septum/diagonal band complex (MS/DB). Slow firing neurons were identified by their low firing rate at threshold (<5 Hz) and lack of time dependent inward rectification (Ih). Unsupervised cluster analysis was used to investigate whether slow firing neurons could be further classified into different subtypes. The parameters used for the cluster analysis included latency for first spike, slow after-hyperpolarizing potential, maximal frequency and action potential (AP) decay slope. Neurons were grouped into three major subtypes. The majority of neurons (55%) were grouped as cluster I. Cluster II (17% of neurons) exhibited longer latency for generation of the first action potential (246.5+/-20.1 ms). Cluster III (28% of neurons) exhibited higher maximal firing frequency (25.3+/-1.7 Hz) when compared with cluster I (12.3+/-0.9 Hz) and cluster II (11.8+/-1.1 Hz) neurons. Additionally, cluster III neurons exhibited faster action potentials at suprathreshold. Interestingly, cluster II neurons were frequently located in the medial septum whereas neurons in cluster I and III appeared scattered throughout all MS/DB regions. Sholl's analysis revealed a more complex dendritic arborization in cluster III neurons. Cluster I and II neurons exhibited characteristics of "true" slow firing neurons whereas cluster III neurons exhibited higher frequency firing patterns. Several neurons were labeled with a cholinergic marker, Cy3-conjugated 192 IgG (p75NTR), and cholinergic neurons were found to be distributed among the three clusters. Our findings indicate that slow firing medial septal neurons are heterogeneous and that soma location is an important determinant of their electrophysiological properties. Thus, slow firing neurons from different septal regions have distinct functional properties, most likely related to their diverse connectivity. PMID- 17412518 TI - An occult risk factor for chronic prostatitis: Helicobacter pylori. PMID- 17412519 TI - Postmenopause is associated with recurrence of differentiated papillary thyroid carcinoma. AB - Differentiated papillary thyroid carcinoma (D-PTC) is the most common malignancy arising in the thyroid gland. There are gender differences in the incidence of PTC being mainly observed in females. Low-risk groups consisted of men younger than 40-year-old and women younger than 50-year-old, whereas the high-risk group are older patients. We believe that age is not enough to explain the clinical course of this neoplasm and hypothesize that aggressive behavior of D-PTC may be correlated with hormonal status. Studies that support this idea showed that the follicular neoplastic cells had higher estrogen receptor-alpha in premenopausal (28.1+/-4.5) than in postmenopausal women (14.2+/-2.9). According to author's prior observations, there are evidences correlating recurrence of D-PTC with postmenopause in women. Postmenopause status is characterized by estrogen decrease and FSH increases both associated with EGFR activation. Previous observations identified EGFR over-expression in D-PTC of postmenopause when compared with premenopausal ladies. CONCLUSIONS: Postmenopause is an adverse factor for tumor evolution in women with D-PTC and is associated with EGFR expression. It's introduction in thyroid tumor stratification could be a fine tuning in predicting papillary thyroid carcinoma behavior. PMID- 17412521 TI - [Dynamic circadian system patients under chimiotherapy]. AB - Currently, the circadian timing system of cancer patients can be estimated with wrist actimetry, which provides numerical data on the continuous activity of patients. The method of processing of these data, which we implemented, aims at answering the following questions: is the circadian timing system stable before chemotherapy, is this system extensively modified by chemotherapy, are there structural modifications or a decrease in activity amplitude produced by chemotherapy and how does the circadian activity rhythm recover? Here, we determine a circadian model function using a wavelet transform prior to chemotherapy delivery. Then we measure the correlation between this model function and the evolution of the circadian rhythm of activity over the whole observation span during and after chemotherapy. The application of this method to the record of activity rhythms in cancer patients highlights the characteristics and the recovery pattern of chemotherapy-induced alterations of the circadian timing system. PMID- 17412520 TI - Autism, asthma, inflammation, and the hygiene hypothesis. AB - Inflammation and the genes, molecules, and biological pathways that lead to inflammatory processes influence many important and disparate biological processes and disease states that are quite often not generally considered classical inflammatory or autoimmune disorders. These include development, reproduction, aging, tumor development and tumor rejection, cardiovascular pathologies, metabolic disorders, as well as neurological and psychiatric disorders. This paper compares parallel aspects of autism and inflammatory disorders with an emphasis on asthma. These comparisons include epidemiological, morphometric, molecular, and genetic aspects of both disease types, contributing to a hypothesis of autism in the context of the immune based hygiene hypothesis. This hypothesis is meant to address the apparent rise in the prevalence of autism in the population. PMID- 17412522 TI - [The endocannabinoid system as a novel target for the treatment of liver fibrosis]. AB - The cannabinoid system comprises specific G protein-coupled receptors (CB1 and CB2), exogenous (marijuana-derived cannabinoids) and endogenous (endocannabinoids) ligands, and a machinery dedicated to endocannabinoid synthesis and degradation. Studies over two decades have extensively documented the crucial role of the cannabinoid system in the regulation of a variety of pathophysiological conditions. However, its role in liver pathology has only been recently unravelled, probably given the low expression of CB1 and CB2 in the normal liver. We have recently demonstrated that CB1 and CB2 receptors display opposite effects in the regulation of liver fibrogenesis during chronic liver injury. Indeed, both receptors are up-regulated in the liver of cirrhotic patients, and expressed in liver fibrogenic cells. Moreover, CB1 receptors are profibrogenic and accordingly, the CB1 antagonist rimonabant reduces fibrosis progression in three experimental models. In keeping with these results, daily cannabis smoking is a risk factor for fibrosis progression in patients with chronic hepatitis C. In contrast, CB2 display antifibrogenic effects, by a mechanism involving reduction of liver fibrogenic cell accumulation. These results may offer new perspectives for the treatment of liver fibrosis, combining CB2 agonist and CB1 antagonist therapy. PMID- 17412523 TI - [Circadian clock gene expression in human peripheral blood mononuclear cells]. AB - Circadian clock genes have been identified in humans but information regarding their expression has remained very limited. However from a basic point as well as in a diagnostic and therapeutic perspective, it is important to evaluate molecular clock gene expression. Peripheral blood mononuclear cells represent an ideal material to investigate non-invasively the human clock at the molecular level. Several studies including ours reported rhythmic expression of clock genes in these cells, with significant intersubject variability of expression. In addition, our results reveal the existence of different chronotypes of clock gene expression patterns and suggest specific regulatory mechanisms in these human cells as compared to other peripheral tissues. PMID- 17412524 TI - [Circadian behaviour of host versus tumour outcome. A review]. AB - The authors reviewed experimental and clinical data emphasizing the importance for host to keep a reference circadian rythmicity. This assessment seems true both in the cancerogenesis phase, as well in the active phase of tumoural disease. PMID- 17412525 TI - [Interactions between clock gene mutation, circadian phenotype and tumor growth in mice]. AB - The relation between circadian physiology (rest-activity and body temperature) and the growth of a grafted tumor (Glasgow osteosarcoma-GOS) was investigated in the mice with mutation of clock gene (ClockDelta19(-)) or gene controlled by the clock (Vpac(-/-)). Circadian rhythms in temperature and activity were stable, with an approximately 24-h period in all the mice synchronized by the alternation of 12 h of light and 12 h of darkness (LD 12:12). Following exposure to constant darkness (DD), both rhythms persisted in ClockDelta19(-), yet with a lengthening of the period by 4.5 h compared to wild type. In DD, the amplitude increased by 45.9% for the temperature rhythm (p<0.001) and by 17.4% for the activity one (p=0.08) as compared to LD 12:12 in ClockDelta19(-). The improvement of circadian coordination and/or the lengthening of the circadian period observed in ClockDelta19(-) kept in DD was associated with a moderate slowing down of tumor growth. Although the exposure to DD ablated the activity and temperature rhythms in Vpac(-/-), no modification in tumor growth was observed as compared to wide type or Vpac(-/-) in LD 12:12. Major alternations of circadian physiology can result from interactions between photoperiodic environment and mutation of clock gene or gene controlled by the clock. In these conditions, we have shown that the alternation of the circadian phenotype does not seem to constitute an essential determinant of the growth of a grafted tumor. PMID- 17412526 TI - [Clock genes, circadian rhythms and food intake]. AB - The molecular clockwork in mammals involves various clock genes with specific temporal patterns of expression. Synchronization of the master circadian clock located in the suprachiasmatic nuclei is accomplished mainly via daily resetting of the phase of the clock by light stimuli. Phase shifting responses to light are correlated with induction of Per1 and Per2 within the suprachiasmatic cells. The timing of peripheral oscillators is controlled by the suprachiasmatic clock when food is available ad libitum. Time of feeding, as modulated by temporal restricted feeding, is a potent Zeitgeber (synchronizer) for peripheral oscillators with no clear synchronizing influence on the suprachiasmatic clockwork. However, a timed calorie restriction (i.e. when only a hypocaloric diet is given each day at the same time) can modify the temporal organization generated by the suprachiasmatic nuclei and reset by the light-dark cycle. Such a situation of conflict between photic and feeding synchronizers alters timing of clock gene expression within the suprachiasmatic nuclei and timing of circadian outputs, indicating that the suprachiasmatic clock is sensitive to nutritional cues. PMID- 17412527 TI - Some conceptual problems with the classical theory of behaviour. AB - Behaviour is usually assumed to depend on the reach of a critical intensity- termed reactivity threshold--by its motivation. This view represents a simple, predictive theoretical framework in ethology and animal psychology. However, it is here argued that only the influence of an isolated motivation on behaviour can be explained that way; that such a view fails to account for behaviour when several motivations are jointly activated. Upon analysis, the classical theory of behaviour (CTB) proves to be under-specified and thus leads to three conceptual problems that make it logically inconsistent for the study of multiple motivations. A revision of the CTB, called anticipatory dynamics model (ADM), is then developed in order to bring a theoretical solution to these conceptual problems. The ADM hypothesizes that an organism's motivational interactions are due to the limitation of the organism's attentional resources. PMID- 17412528 TI - Equine learning behaviour: common knowledge and systematic research. PMID- 17412529 TI - Equine learning behaviour: the importance of evolutionary and ecological approach in research. PMID- 17412530 TI - Short term impact of same intensity but different duration interventions for cannabis users. AB - The present study evaluates the efficacy of a brief intervention for cannabis users. A randomized controlled trial compared 3 conditions: 4 weekly individual sessions of motivational interviewing and relapse prevention over 1 month (1MIRP); the same 4 sessions over 3 months (3MIRP), and delayed treatment control (DTC). The short term impact of each intervention was followed up 4 months after randomization. Participants were 160 highly educated adults with a long history of frequent cannabis use. Both treatments showed better results than the DTC, and for primary outcomes (i.e., cannabis consumption) there was no difference between treatments, while the 3MIRP scheme showed greater efficacy in reducing dependence symptoms and other drug use according to the ASI drug subscale. There was a tendency for the longer treatment to have better outcomes, regardless of intensity, although the waiting list did have some positive effect. The cohort needs to be followed up for a longer period in order to ascertain whether changes are maintained over time. PMID- 17412531 TI - Gene expression in nanotoxicology research: analysis by differential display in BALB3T3 fibroblasts exposed to cobalt particles and ions. AB - Broadly defined, nanoscale materials are substances in which at least one critical dimension is less than 100 nm. Nanoscale materials are employed in several industrial applications as well as in biology and medicine. Despite their wide use, very little research has been carried out on the potential toxicity of nanoparticles. For this reason, we report on a molecular approach in nanotoxicology research. Using the differential display technique, we focused our attention on mRNA expression in a BALB3T3 A31-1-1 cell line that was not exposed and exposed for 72 h to 1 microM of cobalt microparticles (Co-mu), nanoparticles (Co-nano), and ions. In the experiments, we obtained 10 differentially expressed sequences. These genes represent candidate biomarkers capable of indicating specific cellular effects after Co-nano exposure. In addition, our results show that treatment with Co-nano somehow activates cellular pathways of defense and repair mechanisms. It is also evident that molecular techniques are valuable tools in nanotoxicology research, where they will certainly find wide use. PMID- 17412532 TI - Percutaneous absorption and metabolism of 2-butoxyethanol in human volunteers: a microdialysis study. AB - We determined percutaneous absorption kinetics of 2-butoxyethanol (BE) in volunteers using microdialysis. Four male volunteers were dermally exposed twice to 90% and 50% aqueous solutions (v/v) of BE for 4.5h. To determine percutaneous absorption kinetics the concentration of BE was measured in the dialysate samples collected at 30 min-intervals throughout exposure. The systemic absorption, which is needed to determine recovery of the BE in the dialysate, was estimated from the concentration of the main metabolite of BE, free butoxyacetic acid (BAA) in urine. A pseudo steady-state percutaneous absorption was reached approximately at 2h of exposure for both BE concentrations. The maximum dermal flux of 50% BE was higher than that of 90% BE (2.8+/-0.4, 1.9+/-0.6 mg cm(-2)h(-1), respectively). The more diluted BE solution showed shorter lag time: 25 min versus 39 min. The amount of BAA was determined in the pooled dialysate samples collected at 4 and 4.5h. The dermal metabolism seems to be low, the BAA amount ranged from 0.03% to 1.9% of the BE in the same dialysate. Our study demonstrates applicability of microdialysis technique for assessment of percutaneous absorption kinetics and dermal metabolism without interference from the systemic compartment. PMID- 17412533 TI - TGFalpha and EGFR in ovine preimplantation embryos and effects on development. AB - The present study aimed to assess location and relative amounts of transforming growth factor alpha (TGFalpha) and its receptor (EGFR) in ovine oocytes and preimplantation embryos by using immunohistochemical technique that was graded on a relative scale of 0-3, with 0 representing absence of staining, and 3 exhibiting prominent staining, and to evaluate the effects of TGFalpha/EGF on in vitro development of preimplantation embryos by adding different concentrations of EGF and TGFalpha to culture medium. The results showed that EGFR was abundant in cell plasma membranes in immature and mature oocytes, cumulus cells of immature cumulus-oocyte complexes (COC), fertilized oocytes and at different stages of embryo development. However, the relative amounts in inner cell mass (ICM) (1+) was less than that in trophectoderm (TE) cells (2+) at the blastocysts stage. The staining pattern for TGFalpha was a similar to EGFR. However, the staining for TGFalpha slightly increased in the fertilized oocytes (1-2+) as compared to immature and mature oocytes (1+). TGFalpha was mainly detected in the cytoplasm close to the membrane in both ICM and trophectoderm (TE) cells. The developmental rate of 8-cell stage embryos cultured with 5 ng/ml TGFalpha was increased as compared to other treatments (P<0.05). There was no significant difference in the rate of development of blastocysts cultured with 5 ng/ml TGFalpha, 20 ng/ml EGF, 20 ng/ml EGF+5 ng/ml TGFalpha or the control treatment (P>0.05). In addition, there was no significant difference in the number of cells in blastocyst stage as compared with different treatments (P>0.05). However, TGFalpha alone enhanced cell survival rated (P<0.01) and reduced apoptosis. We concluded that TGFalpha can improve development of ovine preimplantation embryos at the 8-cell and blastocyst stages in vitro. PMID- 17412534 TI - Trehalose enhanced the freezability of Poodle dog sperm collected by an artificial vagina (AV). AB - In an attempt to develop a suitable freezing method for Poodle dog sperm, an experiment was conducted to investigate semen collection methods of digital stimulation and an artificial vagina (AV), using Tris and trehalose-egg yolk extender, on the characteristics and cryopreservation of sperm. Two dogs (dogs A and B) were subjected to semen collection by digital stimulation and AV. The volume, sperm concentration, sperm motility index (SMI) and acrosome status of ejaculates were determined immediately after collection. The remainder was frozen as pellets in Tris and trehalose-egg yolk extender. Sperm motility index was evaluated after thawing and during a thermal resistance test, and acrosome integrity was also assessed. No significant differences regarding sperm concentrations, SMI and acrosome integrity were observed between semen collected by AV and digital stimulation. However, when dog sperm were collected by an AV and frozen in trehalose-egg yolk extender, the motility index of frozen-thawed sperm was significantly improved compared to sperm frozen in Tris-egg yolk extender which were collected by digital stimulation. In conclusion, semen collected by an AV and frozen in trehalose-egg yolk extender was effective in enhancing the freezability of Poodle dog sperm. PMID- 17412535 TI - Endometrial bleeding during hormone therapy: the effect of progestogens. AB - Postmenopausal women using continuous combined estrogen/progestin therapy (ccEPT) are likely to have irregular bleeding or spotting. The use of estrogen with 12-14 days of a progestin is called cyclic (scEPT). This method results in regular endometrial bleeding at a scheduled time. The mechanism(s) involved in this regimen that result in bleeding could be similar to a spontaneous menstrual bleeding episode in a menachal woman, but there are no data in this regard. This aspect of regular scheduled bleeding in postmenopausal women will not be addressed in this article due to the paucity of information. The effect of cyclic progestogen with continuous estrogen on the endometrium could result in similar local mechanisms for endometrial bleeding as seen wth ccEPT. The mechanism(s) involved in endometrial bleeding is unknown. Several reports have highlighted a number of potential pathophysiologic mechanisms. Most of the investigation into the mechanisms involved in endometrial bleeding has been in women using progestin only contraceptive methods not ccEPT. The use of ccEPT could be construed as similar but not identical to that of a continuous progestin only contraceptive since the progestin in ccEPT is delivered daily. The potential mechanism(s) involved in endometrial bleeding includes the following: changes in the ratio of vascular endothelial growth factor to Thrombospondin-1 (pro- versus anti angiogenic factors); alterations in metalloproteinases and tissue inhibitor of metalloproteinases (TIMP); changes in tissue factor a known haemostasis mediator in the endometrium; and increased endometrial leukocytes with a particular emphasis on uterine natural killer (uNK) cells. Each of these potential causes has been the subject of both in vivo and in vitro investigations. There is no clear linkage between any of these hypotheses and the onset or cessation of uterine bleeding in ccEPT users. No good therapeutic option to control the bleeding or spotting exists at this time. Evaluation and monitoring of the patient regarding endometrial safety is of paramount importance. PMID- 17412536 TI - All-trans-retinoic acid nanodisks. AB - Nanodisks are nanoscale, disk-shaped phospholipid bilayers whose edge is stabilized by association of apolipoprotein molecules. Self-assembled ND particles enriched with all-trans-retinoic acid (ATRA) (phospholipid:ATRA molar ratio = 5.5:1) were generated wherein all reaction components were solubilized. ATRA-ND migrated as a single band (Stokes' diameter approximately 20 nm) on native gradient polyacrylamide gel electrophoresis. ATRA, phospholipid and apolipoprotein co-eluted from a Sepharose 6B gel filtration column, consistent with stable integration of ATRA into the ND particle milieu. Spectroscopic analysis of ATRA-ND in buffer yielded an absorbance spectrum characteristic of ATRA. ATRA-ND mediated time-dependent inhibition of cultured HepG2 cell growth more effectively than free ATRA. The nanoscale size of the formulation particles and the stable integration of biologically active ATRA suggest ND represent a potentially useful vehicle for solubilization and in vivo delivery of ATRA. PMID- 17412537 TI - The effect of water solubility of solutes on their flux through human skin in vitro: an extended Flynn database fitted to the Roberts-Sloan equation. AB - The edited Flynn database (n=62) for determining the effect of the physicochemical properties of solutes on their skin absorption has been extended (n=114) to give a database for which solubilities of the solutes in water, S(AQ), and their maximum fluxes from water through human skin in vitro, J(MAQ), are known or can be calculated. Besides the six major contributors to the original and edited Flynn database, nine more contributors have been included in the extended database to give 15 contributors. As in the edited Flynn database, data for solutes that were significantly ionized or for experiments using different thicknesses of skin were not excluded from the extended database so that the diversity of the original database was maintained. The extended database was fit to five equations where the independent variables were solubility in octanol (S(OCT)), in water (S(AQ)) or molecular weight (MW) and combinations of those three variables; and the dependent variable was J(MAQ). The best fit was obtained from the Roberts-Sloan (RS) equation: logJ(MAQ) = x + ylogS(OCT) + (1 - y)logS(AQ) - z MW, x = -2.574, y = 0.586, z = 0.00440, r(2) = 0.887, S.D. = 0.399, F = 139. This result is comparable to the best fit published using permeability coefficients, P, as the dependent variable, but gives greater insight into the factors affecting permeation. J(MAQ) is more important clinically because it described how much is permeating per unit area and time, while P is in the units of speed (cmh(-1)). Because of the dependence of J(MAQ) on S(AQ), the selection of new drugs with improved topical delivery should include considerations of their S(AQ) in their design. PMID- 17412538 TI - Levofloxacin implants with predefined microstructure fabricated by three dimensional printing technique. AB - A novel three-dimensional (3D) printing technique was utilized in the preparation of drug implants that can be designed to have complex drug release profiles. The method we describe is based on a lactic acid polymer matrix with a predefined microstructure that is amenable to rapid prototyping and fabrication. We describe how the process parameters, especially selection of the binder, were optimized. Implants containing levofloxacin (LVFX) with predefined microstructures using an optimized binder solution of ethanol and acetone (20:80, v/v) were prepared by a 3D printing process that achieved a bi-modal profile displaying both pulsatile and steady state LVFX release from a single implant. The pulse release appeared from day 5 to 25, followed by a steady state phase of 25 days. The next pulse release phase then began at the 50th day and ended at the 80th day. To evaluate the drug implants structurally and analytically, the microscopic morphologies and the in vitro release profiles of the implants fabricated by both the 3D printing technique and the conventional lost mold technique were assessed using environmental scanning electron microscopy (ESEM) and UV absorbance spectrophotometry. The results demonstrate that the 3D printing technology can be used to fabricate drug implants with sophisticated micro- and macro-architecture in a single device that may be rapidly prototyped and fabricated. We conclude that drug implants with predefined microstructure fabricated by 3D printing techniques can have clear advantages compared to implants fabricated by conventional compressing methods. PMID- 17412539 TI - Excitatory, inhibitory and facilitatory frequency response areas in the inferior colliculus of hearing impaired mice. AB - Individuals with age-related hearing loss often have difficulty understanding complex sounds such as basic speech. The C57BL/6 mouse suffers from progressive sensorineural hearing loss and thus is an effective tool for dissecting the neural mechanisms underlying changes in complex sound processing observed in humans. Neural mechanisms important for processing complex sounds include multiple tuning and combination sensitivity, and these responses are common in the inferior colliculus (IC) of normal hearing mice. We examined neural responses in the IC of C57Bl/6 mice to single and combinations of tones to examine the extent of spectral integration in the IC after age-related high frequency hearing loss. Ten percent of the neurons were tuned to multiple frequency bands and an additional 10% displayed non-linear facilitation to the combination of two different tones (combination sensitivity). No combination-sensitive inhibition was observed. By comparing these findings to spectral integration properties in the IC of normal hearing CBA/CaJ mice, we suggest that high frequency hearing loss affects some of the neural mechanisms in the IC that underlie the processing of complex sounds. The loss of spectral integration properties in the IC during aging likely impairs the central auditory system's ability to process complex sounds such as speech. PMID- 17412540 TI - Early neurological phenotype in 4 children with biallelic PRODH mutations. AB - Hyperprolinemia type I (HPI) results from a deficiency of proline oxidase (POX), involved in the first step in the conversion of proline to glutamate. Diverse phenotypes were described in patients with HPI, prior to the identification of the POX gene (PRODH): whereas various patients were asymptomatic, others had neurological and extraneurological defects. The PRODH gene is located in the region deleted in velocardiofacial syndrome (VCFS). Heterozygous and homozygous mutations have been identified in patients with variable hyperprolinemia and various features (patients with schizophrenia, chromosome 22q11 microdeletions and/or neurological defects). A functional study has divided the PRODH missense mutations into three groups: those leading to mild, moderate, or severe reduction of POX activity. In this study, we report four unrelated children with HPI and a homogeneous severe neurological phenotype. We identified biallelic abnormalities in PRODH in these patients that led to severe reduction of POX activity. These included missense and non-sense mutations, deletions of PRODH and a 22q11 microdeletion. Four other children have been reported with severe biallelic PRODH mutations. The phenotype of these eight patients associates early psychomotor development delay with predominant cognitive defects, autistic features and epilepsy. Their values of hyperprolinemia ranged from 400 to 2200 micromol/L. Patients with biallelic PRODH alterations resulting in severely impaired POX activity had an early onset and severe neurological features. Thus, children with this phenotype and those with a microdeletion in chromosome 22q11, especially those with mental retardation and autistic features, should be tested for hyperprolinemia. Hyperprolinemic patients should be screened for PRODH mutations. PMID- 17412541 TI - [Antibiotherapy for early localized Lyme disease]. AB - OBJECTIVE: The aim of this article is to provide clinicians with guidelines for the antibiotherapy of early-localized Lyme disease. The outcome measures are the clearance of erythema migrans and associated symptoms of early localized Lyme disease and the prevention rate of late complications, with a low risk of adverse effects. Design. The reviewed studies were selected by Medline with the keywords: "erythema migrans, treatment". Sixteen studies comparing treatment or duration of treatment were analyzed. RESULTS: Amoxicillin, doxycycline, and cefuroxim axetil are equally efficacious for early-localized Lyme disease. Azithromycin is an alternative. Most patients respond completely and less than 10% fail to respond. All antibiotics are associated with a low frequency of adverse effects, with the exception of Jarisch Herxheimer reaction which occurs in about 15% of the patients. CONCLUSIONS: We recommend treating adults with amoxicillin (50 mg/kg/day in 3 intakes) or doxycycline (100 mg bid) for 14 days (erythema migrans) to 21 days (early localized Lyme disease with associated symptoms). For children, we recommend amoxicillin (50 mg/kg/day in 3 intakes) or doxycycline (4 mg/kg/day in 2 intakes, maximum 100 mg/dose) above 8 years of age. Cefuroxim axetil (500 mg twice daily for adults or 30 mg/kg/day in 2 intakes, maximum 500 mg/dose, for children), and azithromycin (500 mg/day for adults and 20 mg/kg/day for children for 7-10 days) are second line treatment. PMID- 17412542 TI - Evaluation of intervertebral disc herniation and hypermobile intersegmental instability in symptomatic adult patients undergoing recumbent and upright MRI of the cervical or lumbosacral spines. AB - PURPOSE: The purpose of the study was to determine the difference in findings between recumbent and upright-sitting MRI of the cervical and lumbosacral spine in patients with related sign and symptoms. MATERIALS AND METHODS: A total of 89 patients were studied (lumbosacral spine: 45 patients; cervical: 44 patients). T1 weighted (TR: 350, TE: 20) fast spin echo and T2-weighted (TR: 2500, TE: 160) fast spin echo images were acquired in the sagittal and axial planes in both the recumbent and sitting-neutral positions. The images were acquired on the Upright MRI unit (Fonar Corporation, Melville, NY). Differences were sought between the recumbent and upright-sitting positions at all levels imaged, in both planes. RESULTS: The total number of cases of pathology was 68, including instances of posterior disc herniation and anterior and posterior spondylolisthesis. Focal posterior disc herniations were noted in 55 patients (cervical: 31, lumbosacral: 24) [62% of patients]. Six of these herniations (cervical: 4, lumbosacral: 2) [11%] were seen only on the upright-sitting study. Focal posterior disc herniations were seen to comparatively enlarge in size in 35 patients on the upright-seated examination (cervical: 21, lumbosacral: 14) [72%], and reduce in size in 9 patients (cervical: 5, lumbosacral: 4) [18%]. Degenerative anterior (n: 11) and posterior (n: 2) spondylolisthesis was seen in 13 patients (cervical: 0, lumbosacral: 13) [15% of patient total]. Anterior spondylolisthesis was only seen on the upright-seated examination in 4 patients (cervical: 0, lumbosacral: 4) [31%]. Anterior spondylolisthesis was comparatively greater in degree on the upright-seated study in 7 patients (cervical: 0, lumbosacral: 7) [54%]. Posterior spondylolisthesis was comparatively greater in degree on the recumbent examination in 2 patients (cervical: 0, lumbosacral: 2) [15%]. The overall combined recumbent miss rate in cases of pathology was 15% (10/68). The overall combined recumbent underestimation rate in cases of pathology was 62% (42/68). The overall combined upright-seated underestimation in cases of pathology was 16% (11/69). CONCLUSIONS: Overall, upright-seated MRI was found to be superior to recumbent MRI of the spine in 52 patents (recumbent missed pathology [n: 10]+recumbent underestimated pathology [n: 42]=52/89 total patients: 58%) in cases of posterior disc herniation and anterior spondylolisthesis. This seems to validate the importance of weight-bearing imaging in the spine that might be expected to unmask positional enlarging disc herniations and worsening spondylolisthesis. Overall, recumbent MRI was found to be superior to upright seated MRI in 11 cases (11/89: 12%). The latter finding was possibly due to the fact that upright seated position is actually partial flexion that might be expected to reduce some cases of hypermobile posterior spondylolisthesis. PMID- 17412543 TI - Differential diagnosis of non-functional islet cell tumor and pancreatic carcinoma with sonography. AB - OBJECTIVE: To investigate the differential diagnosis of non-functional islet cell tumor (NFICT) and pancreatic ductal adenocarcinoma (pancreatic carcinoma) with clinical presentation and sonographic features. MATERIALS AND METHODS: Twenty cases of NFICT were investigated in the study, and 41 cases of pancreatic carcinoma were included as the control group. Among them, 5 NFICT and 32 pancreatic carcinomas underwent CEUS with SonoVue. Clinical presentation and sonographic characteristics were evaluated with Logistic regression analysis. Furthermore, the preliminary result of contrast enhanced ultrasound (CEUS) was analyzed. RESULTS: Statistic analysis showed four significant factors in differential diagnosis for NFICT and pancreatic carcinoma, including age (P<0.001), tumor size (P=0.006), tumor margin (P<0.001) and vascularity of tumor (P=0.004). Combined these four factors, the sensitivity, specificity and accuracy are 95.0%, 95.1% and 95.1%, respectively. When the patient is younger than 60 years, and tumor is smaller than 5 cm with well-defined margin and hypervascular, it would be most likely a NFICT other than pancreatic carcinoma. NFICT often shows early enhancement and more homogeneous infusion than pancreatic carcinoma on CEUS (P=0.005 and 0.008). CONCLUSIONS: Sonography is able to provide useful differential information for NFICT, which is often misdiagnosed as pancreatic carcinoma. PMID- 17412544 TI - Detection of intracranial aneurysms with 64 channel multidetector row computed tomography: comparison with digital subtraction angiography. AB - PURPOSE: To compare the contribution of 64 channel multidetector row computed tomography angiography (64MDCT-angiography) with digital subtraction angiography (DSA) in the detection of intracranial aneurysms. METHODS AND MATERIALS: Twenty nine patients (10 males and 19 females, age: 40-84 years; average: 61.9 years) with clinical and imaging findings strongly suggesting the presence of subaracnoid hemorrhage underwent 64MDCT-angiography and DSA with a short interval between the two examinations (less than 12 h-5 days). CT parameters were: 64 mm x 0.5 mm collimation, pitch-0.828 and helical pitch-53. DSA were performed with standard technique (four vessel catheterization) and multiple projections. Axial CT scans as well as maximum intensity projection, volume rendering and multiplanar reformations and angiographic views were independently reviewed by four readers (two for 64MDCT-angiography and two for DSA). Consensus was reached for discordant cases. DSA was considered as the standard of reference. RESULTS: In 29 patients, 28 aneurysms were found (14 patients had 1 aneurysm, 4 patients had 2 aneurysms and 2 patients had 3 aneurysms; in 9 patients no aneurysm were found). 64MDCT-angiography detected 26/28 aneurysms. No false-positive sites were recognized. The sensitivity, specificity, positive and negative predictive values and diagnostic accuracy were, respectively, 92.8, 100, 100, 99.4 and 99.5%. CONCLUSIONS: 64MDCT-angiography is helpful in detecting intracranial aneurysms with results similar to those of DSA but with less discomfort and risks for the patients and can be considered for the first line imaging technique. Conventional angiography is still needed in doubtful cases or negative MDCT-angiography associated with a strong clinical suspect. PMID- 17412545 TI - The diagnostic role of abdominal CT imaging findings in adults intussusception: focused on the vascular compromise. AB - Intussusception is defined as telescoping of one segment of the gastrointestinal tract into an adjacent one. Unlike that in children, adult intussusception is a relatively rare condition. More than 90% of patients with adult intussusception have been reported to have an organic cause, with benign or malignant tumors for accounting for approximately 65% of the cases. In general, the diagnosis is easily made by means of computed tomography (CT) or magnetic resonance (MR) imaging. The imaging appearance of a bowel-within-bowel configuration with or without contained fat and mesenteric vessels, is pathognomonic. As the intussusceptum enters into the intussuscipiens, the mesentery is carried forward and trapped between the overlapping layers of bowel. The twisting or severe constriction of the mesenteric vessels may result in vascular compromise with subsequent edematous thickening of the involved bowel. In these circumstances, ischemic necrosis may develop if timely intervention is not undertaken. Therefore, determination of the presence or absence of intestinal necrosis in intussusception is important in patient management. On CT, the presence of well known diagnostic CT criteria for strangulated obstruction (especially severe engorgement or twisting of the mesenteric vessels) as well as evidence of loss of the layered pattern, accumulation of extraluminal fluid collection, and bowel perforation, may suggest the diagnosis of intestinal necrosis. CT and MR imaging are limited in determining the primary disease causing intussusception. However, CT and MR provide excellent pre-operative evaluation, including the possible extension and/or dissemination of a malignant tumor. CT and MR imaging may also be useful in suggesting the presence of vascular compromise. PMID- 17412546 TI - Efficacy and safety of gadodiamide (Gd-DTPA-BMA) in renal 3D-magnetic resonance angiography (MRA): a phase II study. AB - PURPOSE: To determine the most efficacious dose of gadodiamide for three dimensional (3D) contrast-enhanced (CE) magnetic resonance angiography (MRA) of the renal arteries on a patient level based on the sensitivity in detecting the main hemodynamically relevant (> or =50% or occlusion) renal artery stenosis (RAS) using intra-arterial digital subtraction angiography (IA DSA) as the gold standard. MATERIALS AND METHODS: This prospective, randomized, double-blind, parallel-group, multicenter study included 273 patients referred to IA DSA for suspected RAS. Patients underwent 3D CE MRA after injection of 0.01, 0.05, 0.1, or 0.2mmol/kg of body weight gadodiamide (0.5mmol/ml). The images were assessed for location and degree of RAS by independent blinded readers (MRA: three readers, IA DSA: one reader). Hypothesis testing for a significant trend in sensitivity across dose groups was based on the one-sided Cochran-Armitage style trend test for each independent MRA reader. RESULTS: The lowest dose group (0.01mmol/kg) proved non-efficacious in detecting hemodynamically relevant (i.e., > or =50% or occlusion) RAS. A statistically significant dose trend (p<0.001) was shown for each of the three independent readers. Depending on reader, the sensitivity obtained with 0.05, 0.1, and 0.2mmol/kg was 63.9-86.1%, 75.8-91.4% and 80.6-90.6%, the specificity was 66.7-73.9%, 59.3-75.0%, and 59.3-75.0% and accuracy was 67.8-78.9%, 75.4-77.4%, and 76.3-81.0%, for the three dose groups, respectively. There were eight non-severe adverse events (AEs). Three serious AEs occurring in one patient were judged not related to gadodiamide by the on-site investigator. CONCLUSION: A significant dose trend between the four doses examined was observed. The lowest dose (0.01mmol/kg) differed significantly from those of the other three doses. Based on the analysis of the primary and secondary endpoints, 0.1mmol/kg gadodiamide appears to be the most suitable dose in diagnosing hemodynamically relevant RAS. The present study also demonstrated gadodiamide to be safe and well tolerated. PMID- 17412547 TI - Identification of volatile degradants in formulations containing sesame oil using SPME/GC/MS. AB - Solid-phase microextraction (SPME), in combination with gas chromatography/mass spectrometry (GC/MS), was used to identify an unknown degradant observed during stability studies of a pharmaceutical formulation containing sesame oil. SPME is a solvent-less, rapid, sensitive, and inexpensive extraction method that minimizes sample preparation. SPME combined with GC is a widely used technique in certain fields, such as food, environmental analysis, forensics, and consumer products, but has only rarely been used for the analysis of pharmaceutical formulations. Hexanal, octanal, 2-octenal, 2-decenal, 2-undecenal, and 2,4 decadienal can be detected and identified by GC/MS, but they cannot be detected by LC/MS due to their volatility and low ionization efficiency under atmospheric pressure ionization conditions. Combining the MS data from the GC/MS with LC/DAD data resulted in the identification of the unknown degradant in the formulation as 2,4-decadienal. The presence of this and other aldehydes was attributed to the oxidative degradation of the unsaturated fatty-acid component in vegetable oils. PMID- 17412548 TI - The safety and efficacy of radiofrequency and electrolytic ablation created adjacent to large hepatic veins in a porcine model. AB - INTRODUCTION: Immediately adjacent to large hepatic veins, tumour ablation by radiofrequency or electrolysis may be impaired by heat or current sink effects. Ablation may also cause vessel injury and thrombosis. The aim of this study was to evaluate the safety and efficacy of radiofrequency and electrolytic ablative techniques adjacent to large hepatic veins. METHODS: Electrolytic and radiofrequency zones of ablation were created adjacent to hepatic veins in large white pigs. After 72 h the zones of ablation created were examined histologically for (a) the extent of tissue necrosis up to the vessel and (b) the presence of intimal damage and mural thrombus in the veins. RESULTS: An unexpected complication of electrolysis near large veins was cardiac tamponade. This current related phenomenon could easily be avoided. In seven of nine electrolysis zones of ablation necrosis was completely adjacent to the vessel wall, but in only four of seven radiofrequency zones of ablation. All zones of ablation were associated with intimal necrosis, and most with mural thrombosis. CONCLUSIONS: Ablation of hepatic tumours by radiofrequency and electrolysis is unreliable adjacent to hepatic veins. Both techniques are associated with mural thrombus formation, and so risk thrombo-embolic complication. These ablative modalities are not recommended for zones of ablation adjacent to hepatic veins. PMID- 17412549 TI - [Be aware of wrong conclusions!]. PMID- 17412550 TI - Therapeutic adherence to bisphosphonates. AB - Therapeutic adherence of patients is a key factor of treatment success in clinical praxis, although it is often neglected. Several studies have shown that insufficient persistence and compliance cause differences in the efficiency of treatments in clinical studies and clinical praxis. A recent meta-analysis even showed a clear inverse relationship between therapeutic adherence and mortality. Factors influencing the adherence to treatment include explanations by the physician, characteristics of the disease, patient's attitudes, but also the therapeutic regime. Osteoporosis as a chronic disorder with relatively long asymptomatic initial course represents a major problem. In addition, the currently available therapeutic regimes are discomfortable and, thus, contribute to the low therapeutic adherence of the patient. One of the factors causing discomfort in bisphosphonates therapy is the frequency of application--once daily or once weekly. Several questionnaire-based studies have shown that patients clearly prefer the new alternative once monthly regime available for ibandronate. Although the efficiency of the drug is proven in large clinical trials, the effects of the once monthly regime itself on hard clinical end-points like mortality can only be analyzed in long-term follow-up studies. PMID- 17412551 TI - Enhanced antitumor activity of irinotecan combined with propolis and its polyphenolic compounds on Ehrlich ascites tumor in mice. AB - The effects of the anticancer drug irinotecan combined with ethanolic extract of propolis (EEP), a water-soluble derivate of propolis (WSDP), quercetin and naringin on the growth of Ehrlich ascites tumor (EAT) and the life span of tumor bearing Swiss albino mice were studied. Test components were given to mice intraperitoneally (i.p.) at doses of 100mg kg(-1) for three consecutive days before the i.p. injection of EAT cells (1x10(6)). Irinotecan was administered i.p. at dose of 50mg kg(-1) on days 1, 13, and 19 after tumor cell inoculation. The results clearly demonstrate the synergistic action of irinotecan and EEP on survival time. These results suggest that clinical trials using a propolis preparation EEP combined with irinotecan may be beneficial in maximizing antitumor activity and minimizing post-chemotherapeutic reactions to the cytostatic drug. PMID- 17412552 TI - Poly-N-acetylglucosamine mediates biofilm formation and antibiotic resistance in Actinobacillus pleuropneumoniae. AB - Most field isolates of the swine pathogen Actinobacillus pleuropneumoniae form tenacious biofilms on abiotic surfaces in vitro. We purified matrix polysaccharides from biofilms produced by A. pleuropneumoniae field isolates IA1 and IA5 (serotypes 1 and 5, respectively), and determined their chemical structures by using NMR spectroscopy. Both strains produced matrix polysaccharides consisting of linear chains of N-acetyl-D-glucosamine (GlcNAc) residues in beta(1,6) linkage (poly-beta-1,6-GlcNAc or PGA). A small percentage of the GlcNAc residues in each polysaccharide were N-deacetylated. These structures were nearly identical to those of biofilm matrix polysaccharides produced by Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis. PCR analyses indicated that a gene encoding the PGA-specific glycoside transferase enzyme PgaC was present on the chromosome of 15 out of 15 A. pleuropneumoniae reference strains (serotypes 1-12) and 76 out of 77 A. pleuropneumoniae field isolates (serotypes 1, 5 and 7). A pgaC mutant of strain IA5 failed to form biofilms in vitro, as did wild-type strains IA1 and IA5 when grown in broth supplemented with the PGA-hydrolyzing enzyme dispersin B. Treatment of IA5 biofilms with dispersin B rendered them more sensitive to killing by ampicillin. Our findings suggest that PGA functions as a major biofilm adhesin in A. pleuropneumoniae. Biofilm formation may have relevance to the colonization and pathogenesis of A. pleuropneumoniae in pigs. PMID- 17412553 TI - E-cadherin is a receptor for the common protein pneumococcal surface adhesin A (PsaA) of Streptococcus pneumoniae. AB - Streptococcus pneumoniae (Pnc) binds to nasopharyngeal (NP) epithelial cells in the first steps of nasopharyngeal carriage and colonization through bacterial adhesins. The pneumococcal surface adhesin A (PsaA) has previously been reported to play a significant role in pneumococcal adherence and colonization. Identification of a receptor for PsaA on human epithelium will aid in understanding the pathogenesis of this bacterium. Using recombinant PsaA covalently bound to fluorescent spheres (fluospheres), we show PsaA binds to NP cells through interaction with the human cellular receptor, E-cadherin. SDS-PAGE silver stain analysis demonstrates binding of PsaA to E-cadherin. Recombinant human E-cadherin binds to and blocks PsaA-coated fluospheres and whole transparent bacteria from adhering to NP cells, but does not block a Pnc PsaA(-) mutant. Recombinant E-selectin and human alpha(5)beta(1) integrin did not bind to or block PsaA-coated fluosphere adherence to NP cells. Likewise, if NP cells were preincubated with anti-E-cadherin antibody, there was a significant decrease (46%, P=0.05) in PsaA-coated fluosphere adherence to the cells. Additionally, when using E-cadherin transfected cells, we observed PsaA-coated fluospheres bind more efficiently to cells which express E-cadherin. This work identifies E cadherin as a receptor on human epithelial cells for the pneumococcal surface adhesin, PsaA. PMID- 17412554 TI - Influence of endogenous and synthetic female sex hormones on human blood cells in vitro studied with comet assay. AB - The comet assay has been conducted with numerous cell lines to assess in vitro genotoxicity. In order to use the comet assay as part of an in vitro test for evaluating genotoxicity, however, there are cell-specific factors that need to be better understood. In this present study we have evaluated some factors that may impact upon the DNA damage detected in whole blood (WB) cells and lymphocytes (ILs). Experiments were conducted comparing responses of both cells, and investigating the effects of the female hormonal cycle, and oral contraceptive (OC) use on DNA damage detection in the in vitro comet assay, at three sampling time. No significant differences were detected in the basal levels of DNA damage detected in ILs and WB cells from women OC users and non-users and from men. Basal DNA damage in ILs was unaffected by gender and stage of the menstrual cycle or the stage of the treatment schedule. Our results also indicated that the H2O2 induces DNA damage in human lymphocytes independently of gender, low-dose OC use and hormonal fluctuation. However, data showed that in 3rd sampling of menstrual cycle, lymphocytes were more resistant to H2O2-induced DNA damage than those from OC users and men. PMID- 17412555 TI - Effects of phenytoin and carbamazepine on calcium transport in Caco-2 cells. AB - Adverse effects of anti-seizure/anti-epileptic medications on bone density have been observed and reported since the early 1960s. Phenytoin and carbamazepine are two commonly prescribed anti-epileptic drugs most frequently associated with osteomalacia including fractures, bone demineralization, and reduced bone formation. The mechanism by which anti-epileptic drugs induce bone loss is not fully explained. We hypothesized that anti-epileptic drugs may impair dietary calcium absorption in the intestine. Using Caco-2 cells, a model transport system for study of the function of the intestinal epithelium, we determined the effects of several anti-epileptic drugs on intestinal epithelial calcium transport. In our system, phenytoin and carbamazepine dose-dependently inhibit active calcium transport from the apical to basolateral side of Caco-2 cells under physiologic calcium conditions. Vitamin D ameliorates the anti-epileptic drug-induced decrease in calcium permeability. PMID- 17412556 TI - Variations in anticipatory cognitive stress appraisal and differential proinflammatory cytokine expression in response to acute stress. AB - OBJECTIVE: Anticipatory cognitive appraisal can affect the stress-induced release of stress hormones and stress hormones can modulate monocyte cytokine expression. We investigated whether anticipatory cognitive appraisal processes would predict changes in monocyte cytokine expression following psychosocial stress in relation to stress hormone release. METHODS: Forty-four men (mean age 43+/-2 years; mean arterial blood pressure (MAP) 102+/-2 mmHg; mean body mass index (BMI) 26+/ .4kg/m(2)) completed the Primary Appraisal Secondary Appraisal (PASA) scale before undergoing the Trier Social Stress Test (combination of mock job interview and mental arithmetic task). Lipopolysaccharide (LPS)-stimulated tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 expression by monocytes was assessed in vitro immediately before and after stress, and during recovery up to 60min post stress. Moreover, we repeatedly measured salivary cortisol as well as plasma epinephrine and norepinephrine levels. RESULTS: Stress hormones increased and cytokines decreased following stress (all p<0.05). Correlation analyses showed that a higher PASA "stress index" was associated with higher expression (area under the curve, AUC) of total LPS-stimulated TNF-alpha (r=.33, p=.03) and IL-6 (r=.32, p=.33) between rest and 60min post-stress. While controlling for age, BMI, and MAP, higher values in the primary PASA scale "control expectancy" predicted lower TNF-alpha expression following stress (ss=-.42, p=0.003). Higher "control expectancy" (ss=-.32, p=0.031) and lower "challenge" (ss=.30, p=0.046) predicted lower IL-6 expression. None of the stress hormones predicted expression of any cytokine. CONCLUSIONS: We found that anticipatory cognitive stress appraisal modulates monocyte inflammatory activity following stress suggesting that interventions aiming at improving coping skills might modify the monocyte cytokine response. PMID- 17412557 TI - Thymus atrophy during Trypanosoma cruzi infection is caused by an immuno endocrine imbalance. AB - C57BL/6 mice infected with Trypanosoma cruzi, the causal agent of Chagas' disease, develop severe thymocyte depletion paralleled by an inflammatory syndrome mediated by tumor necrosis factor-alpha (TNF-alpha). The exacerbated inflammatory reaction induces the activation of hypothalamus-pituitary-adrenal (HPA) axis with the consequent release of corticosterone (CT) into the circulation as a protective response. Thymocyte apoptosis has been related to a rise in TNF-alpha and CT levels, and both mediators are increased in T. cruzi infected C57BL/6 mice. The depletion of immature CD4(+)CD8(+) thymocytes by apoptosis following infection with the parasite was still present in mice defective in both types of TNF-receptors (double knockout). However, thymic atrophy was prevented by adrenalectomy combined with RU486 administration, demonstrating that this is a CT-driven phenomenon. Our results put emphasis on the importance of an appropriated immuno-endocrine balance during T. cruzi infection and show that functional deviations in the immuno-endocrine equilibrium have profound effects on the thymus and disease outcome. PMID- 17412558 TI - Pre-natal exposures to cocaine and alcohol and physical growth patterns to age 8 years. AB - Two hundred and two primarily African American/Caribbean children (classified by maternal report and infant meconium as 38 heavier, 74 lighter and 89 not cocaine exposed) were measured repeatedly from birth to age 8 years to assess whether there is an independent effect of pre-natal cocaine exposure on physical growth patterns. Children with fetal alcohol syndrome identifiable at birth were excluded. At birth, cocaine and alcohol exposures were significantly and independently associated with lower weight, length and head circumference in cross-sectional multiple regression analyses. The relationship over time of pre natal exposures to weight, height, and head circumference was then examined by multiple linear regression using mixed linear models including covariates: child's gestational age, gender, ethnicity, age at assessment, current caregiver, birth mother's use of alcohol, marijuana and tobacco during the pregnancy and pre pregnancy weight (for child's weight) and height (for child's height and head circumference). The cocaine effects did not persist beyond infancy in piecewise linear mixed models, but a significant and independent negative effect of pre natal alcohol exposure persisted for weight, height, and head circumference. Catch-up growth in cocaine-exposed infants occurred primarily by 6 months of age for all growth parameters, with some small fluctuations in growth rates in the preschool age range but no detectable differences between heavier versus unexposed nor lighter versus unexposed thereafter. PMID- 17412559 TI - Reinforcement learning for a biped robot based on a CPG-actor-critic method. AB - Animals' rhythmic movements, such as locomotion, are considered to be controlled by neural circuits called central pattern generators (CPGs), which generate oscillatory signals. Motivated by this biological mechanism, studies have been conducted on the rhythmic movements controlled by CPG. As an autonomous learning framework for a CPG controller, we propose in this article a reinforcement learning method we call the "CPG-actor-critic" method. This method introduces a new architecture to the actor, and its training is roughly based on a stochastic policy gradient algorithm presented recently. We apply this method to an automatic acquisition problem of control for a biped robot. Computer simulations show that training of the CPG can be successfully performed by our method, thus allowing the biped robot to not only walk stably but also adapt to environmental changes. PMID- 17412560 TI - Converting antigen-specific diabetogenic CD4 and CD8 T cells to TGF-beta producing non-pathogenic regulatory cells following FoxP3 transduction. AB - Immuno-regulatory defects, including a reduction in the number and function of regulatory T cells, play an important role in the development of autoimmune diabetes in both humans and non-obese diabetic (NOD) mice. In this study we tested the effect of introduction of FoxP3 into antigen non-specific polyclonal and antigen-specific monoclonal T cells on diabetes development in NOD mice. Transduction of FoxP3 into antigen-specific monoclonal (insulin or BDC2.5 mimotope specific) or antigen non-specific polyclonal T cells using retroviral transduction delayed or prevented diabetes development. However, transduced antigen-specific monoclonal T cells were considerably more effective than polyclonal T cells. Regulatory activity was not limited to CD4 T cells as potent diabetogenic CD8 T cells specific for insulin, were also reduced in pathogenicity by FoxP3 induction. The disease suppressive effect, in both CD4 and CD8 cells, was more evident in spontaneously diabetes-prone NOD hosts (non-lymphopenic) than in lymphopenic NOD.scid hosts. We suggest that this strategy of transducing antigen-specific CD4 or CD8 T cells may be a useful therapeutic approach in the prevention of autoimmune diabetes. PMID- 17412561 TI - Persistent seizures following left temporal lobe surgery are associated with posterior and bilateral structural and functional brain abnormalities. AB - PURPOSE: To perform a quantitative MRI and retrospective electrophysiological study to investigate whether persistent post-surgical seizures may be due to brain structural and functional abnormalities in temporal lobe cortex beyond the margins of resection and/or bilateral abnormalities in patients with temporal lobe epilepsy (TLE). METHODS: In 22 patients with left TLE and histopathological evidence of hippocampal sclerosis, we compared pre-surgical brain morphology between patients surgically remedied (Engel's I) and patients with persistent post-surgical seizures (PPS, Engel's II-IV) using voxel-based morphometry (VBM). Routine pre-surgical EEG and invasive and non-invasive telemetry investigations were additionally compared between patient groups. RESULTS: Results indicated widespread structural and functional abnormalities in patients with PPS relative to surgically remedied patients. In particular, patients with PPS had significantly reduced volume of the ipsilateral posterior medial temporal lobe and contralateral medial temporal lobe relative to surgically remedied patients. Furthermore, successful surgery was associated with clear anterior (89%) and unilateral (100%) temporal lobe EEG abnormalities, whilst PPS were associated with widespread ipsilateral (91%) and bilateral (82%) temporal lobe abnormalities. DISCUSSION: We suggest that these preliminary data support the hypothesis that PPS after temporal lobe surgery are due to functionally connected epileptogenic cortex remaining in the ipsilateral posterior temporal lobe and/or in temporal lobe contralateral to resection. PMID- 17412562 TI - NADPH diaphorase reactive neurons in temporal lobe cortex of patients with intractable epilepsy and hippocampal sclerosis. AB - Several studies have demonstrated a controversial involvement of NO in epileptogenesis. The aim of this study is to compare the NADPH diaphorase (NADPH d) reactivity in the temporal cortex between surgical specimens of patients with intractable epilepsy and hippocampal sclerosis and autopsy controls. Brain samples of patients and postmortem controls were stained with the NADPH-d technique. Sprouting and larger areas of NADPH-d reactive neurons were found in the temporal cortex of epileptic patients. PMID- 17412563 TI - Systematic study of association of four GABAergic genes: glutamic acid decarboxylase 1 gene, glutamic acid decarboxylase 2 gene, GABA(B) receptor 1 gene and GABA(A) receptor subunit beta2 gene, with schizophrenia using a universal DNA microarray. AB - Several studies have suggested the dysfunction of the GABAergic system as a risk factor in the pathogenesis of schizophrenia. In the present study, case-control association analysis was conducted in four GABAergic genes: two glutamic acid decarboxylase genes (GAD1 and GAD2), a GABA(A) receptor subunit beta2 gene (GABRB2) and a GABA(B) receptor 1 gene (GABBR1). Using a universal DNA microarray procedure we genotyped a total of 20 SNPs on the above four genes in a study involving 292 patients and 286 controls of Chinese descent. Statistically significant differences were observed in the allelic frequencies of the rs187269C/T polymorphism in the GABRB2 gene (P=0.0450, chi(2)=12.40, OR=1.65) and the -292A/C polymorphism in the GAD1 gene (P=0.0450, chi(2)=14.64 OR=1.77). In addition, using an electrophoretic mobility shift assay (EMSA), we discovered differences in the U251 nuclear protein binding to oligonucleotides representing the -292 SNP on the GAD1 gene, which suggests that the -292C allele has reduced transcription factor binding efficiency compared with the 292A allele. Using the multifactor-dimensionality reduction method (MDR), we found that the interactions among the rs187269C/T polymorphism in the GABRB2 gene, the -243A/G polymorphism in the GAD2 gene and the 27379C/T and 661C/T polymorphisms in the GAD1 gene revealed a significant association with schizophrenia (P<0.001). These findings suggest that the GABRB2 and GAD1 genes alone and the combined effects of the polymorphisms in the four GABAergic system genes may confer susceptibility to the development of schizophrenia in the Chinese population. PMID- 17412564 TI - Cystatin A suppresses ultraviolet B-induced apoptosis of keratinocytes. AB - BACKGROUND: Cystatin A is a cysteine proteinase inhibitor abundantly expressed in keratinocytes. Although cystatin A is one of the cornified cell envelope constituents and expressed in the upper epidermis, its precise function is still unknown. Ultraviolet B irradiation (UVB) induces apoptosis accompanied with the activation of cysteine proteinases, caspases. OBJECTIVE: We investigated the effect of cystatin A on UVB-induced apoptosis of keratinocytes. METHODS: We assessed the caspase activities and apoptotic cell numbers induced by UVB ittadiation in cystatin A gene transfected keratinocytes. RESULTS: UVB-induced pro-caspase 3 cleavage and caspase 3 activation were suppressed in cystatin A expression vector-transfected SV40-transformed human keratinocytes (SVHK). Furthermore, the transfected SVHK cells were resistant to UVB-induced apoptosis. In contrast neither caspase 8 nor caspase 9 activities were affected by UVB irradiation in cystatin A-transfected SVHK cells. The effects were also observed in cystatin A expression adenovirus vector-transfected cultured normal human keratinocytes (NHK). Conversely knockdown of cystatin A by si-RNA induced marked apoptosis of NHK cells following UVB irradiation accompanied with increased caspase 3 activity. In order to confirm the antiapoptotic effect of cystatin A in vivo UVB irradiation was performed on cystatin A transgenic mice (cystatin A-tg). The epidermis from cystatin A-tg was resistant to UVB-induced apoptosis compared to control mice epidermis. CONCLUSION: These results indicate that cystatin A suppresses UVB-induced apoptosis of keratinocytes by the inhibition of caspase 3 activation. PMID- 17412565 TI - Analysis of perforin expression in peripheral blood and lesions in severe and mild psoriasis. AB - BACKGROUND: Perforin is a membrane-disrupting protein that allows the entry of granzymes into a target cell inducing degradation of target substances in the cytoplasm and nucleus thus leading to programmed cell death or apoptosis. Recent work demonstrated a possible involvement of perforin mediated cytotoxicity in immunopathogenesis of psoriasis. OBJECTIVES: To investigate a difference in systemic (peripheral blood) and local (lesions) expression and distribution of perforin in psoriatic patients with severe and mild disease. METHODS: Flow cytometry was used for simultaneous detection of intracellular (perforin) and cell surface antigens in peripheral blood lymphocytes. The expression of perforin in skin lesions was evaluated by immunohistochemistry. RESULTS: Significant increase of perforin expression in T lymphocytes, especially cytotoxic CD8+ cells was found in severe psoriasis compared to mild disease (p<0.01 and p<0.05, respectively). There was also an increase of CD56+P+ NK cells (p<0.05) in severe compared to mild psoriasis. The psoriatic plaque of both, severe and mild disease were abundant with perforin showing no significant difference on local level. CONCLUSION: Based on our results we suggest the association between perforin expression and disease severity. PMID- 17412566 TI - Generic substitution of itraconazole resulting in sub-therapeutic levels and resistance. PMID- 17412567 TI - Neural correlates of trauma script-imagery in posttraumatic stress disorder with and without comorbid major depression: a functional MRI investigation. AB - The goal of this study was to compare neural activation patterns in patients with PTSD with and without current comorbid major depression. Traumatized subjects with PTSD (n=11), PTSD+major depression (MDD, n=15), and subjects (n=16) who met criterion A for PTSD but never developed the disorder were studied using the script-driven symptom-provocation paradigm adapted to functional magnetic resonance imaging (fMRI) at a 4-Tesla field strength. Both the PTSD+MDD and PTSD MDD groups revealed decreased brain activation in the anterior cingulate gyrus (BA 24) and the right ventrolateral prefrontal cortex (BA 47). After covariation for differences in PTSD severity between these groups, the left insula (BA 13) remained more significantly activated in the PTSD-MDD group than in the PTSD+MDD group. In contrast, the PTSD+MDD group showed greater activation than the PTSD MDD group in the bilateral anterior cingulate gyrus (BA 24) and posterior cingulate cortices (BA 23, 31). These results suggest different patterns of brain activation related to comorbid major depression occurring in the context of PTSD. PMID- 17412568 TI - Heteronuclear dipolar recoupling using Hartmann-Hahn cross polarization: a probe for 19F-13C distance determination of fluorinated carbon materials. AB - A NMR determination of the C-F bond length in fluorinated carbon materials using dipolar recoupling is described. To this end Hartmann-Hahn cross polarization with magic angle spinning (inverse cross polarization sequence) is used. A description of the corresponding 13C magnetization evolution as a function of the evolution time and its simulation for typical fluorinated samples are realized. The procedure is applied to 15 different materials having different bonding (semi covalent or covalent) and from various carbon allotropic varieties. The distance evolves from 0.138+/-0.002 nm for covalent bonding to 0.1445+/-0.002 nm for semi covalent bonding. Other parameters may affect the C-F bond length e.g. steric hindrance which leads for fluorinated fullerenes to an increase of this distance up to 0.146+/-0.002 nm. PMID- 17412569 TI - An ensemble of paired spin(-1/2) nuclei in a rotating solid: Polarization evolution and NMR spectrum in a wobbling frame. AB - The evolution of the magnetic polarization of an ensemble of paired spin(-1/2) nuclei in an MAS NMR (nuclear magnetic resonance) experiment and the induced spectrum are described theoretically by means of a Liouville-von Neumann equation representation in a wobbling rotating frame in combination with the averaged Hamiltonian theory. In this method, the effect of a high-intensity external static magnetic field and the effects of the leftover interaction components of the Hamiltonian that commute with the approximate Hamiltonian are taken into account simultaneously and equivalently. This method reproduces details that really exist in the recorded spectra, caused by secular terms in the Hamiltonian, which might otherwise be smoothed out owing to the approximate treatment of the effects of the secular terms. Complete analytical expressions, which describe the whole NMR spectrum including the rotational sideband sets, and which consider all the relevant intermolecular interactions, are obtained. PMID- 17412570 TI - Cross polarization via the non-Zeeman spin reservoirs under MAS. PMID- 17412571 TI - Evidence for vesicle formation from 1:1 nonionic surfactant span 60 and fatty alcohol mixtures in aqueous ethanol: potential delivery vehicle composition. AB - A study of the self-organization of nonionic surfactant span 60 (sorbitan mono stearate) in presence of fatty alcohol (stearyl, cetyl and lauryl) is presented. When ethanolic solution of the surfactant-fatty alcohol (1:1) mixture is added in water spontaneous large unilamellar vesicles (LUV) are formed which may potentially be useful vehicles for drug delivery purposes. Vesicular suspension has been characterized by transmission electron microscopy, dynamic light scattering, confocal laser scanning microscopy, dye entrapment and release studies. Surface tension measurement indicates the suitability of fatty alcohols towards spontaneous vesicle formation from span 60. PMID- 17412572 TI - [Failure to thrive and psychomotor regression revealing vitamin B12 deficiency in 3 infants]. AB - The newborn's vitamin B12 storage exclusively comes from placenta transfer, later from animal food. We relate 3 observations of infants (3-11-13 months) with failure to thrive, anorexia, vomiting and for the two olders refusal of weaning, associated with psychomotricity regression and hypotony. Blood cell count showed a macrocytosis without anemia (case 2-3) and a severe microcytic anemia for the first case caused by a mild alpha-thalassemia, with megaloblastic bone marrow. Vitamin B12 levels were very low associated with increased methylmalonic acid and homocysteine serum levels which confirm the diagnostic . Cerebral imaging showed diffuse cortical atrophy. Cobalamin deficiency was caused by strict vegetarian diets mothers of breastfed infants (cases 2-3) and for younger by mother's unrecognized pernicious anemia. 3 mothers had no anemia and normal B12 's levels at diagnosis. Vitamin B12 supply lead to a rapid clinical and hematologic improvement. In two cases, neurologic recovery was incomplete. About one hundred case of B12 deficiency 's infant are reported, 2/3 are breast-fed by vegetarian mothers, and 1/4 have mothers with pernicious anemia. The failure to thrive is due to anorexia, refusal of weaning and partial villous atrophy. Neurologic manifestations are secondary to cerebral disorders, sometimes revealed by an exposure to anesthetic nitrous oxyd. The macrocytic anemia is inconstant. The etiologic research of developmental delay in an infant may include vitamin B12's deficiency, even if there is no haematologic signs, especially if breast-fedding 's mothers is vegetarian. PMID- 17412573 TI - [Post-traumatic lipid inclusion cyst]. PMID- 17412574 TI - The relationship of gamma-glutamyltransferase to C-reactive protein and arterial stiffness. AB - BACKGROUND AND AIMS: The relationships between gamma-glutamyltransferase (GGT), C reactive protein (CRP), and arterial stiffness have not been fully investigated. The aim of this study was to clarify whether serum GGT is related to CRP and arterial stiffness estimated using brachial-ankle pulse wave velocity (baPWV). METHODS AND RESULTS: The subjects were 3412 males and 854 females. GGT, CRP, baPWV, and conventional risk factors were evaluated. On multiple regression analysis, after adjustment for the conventional risk factors, log GGT was significantly associated with log CRP in male and female subjects (male subjects: beta=0.168, p<0.0001; female subjects: beta=0.098, p<0.05). After adjustment for the conventional risk factors, log GGT was significantly associated with PWV in male subjects (beta=0.060, p<0.0001), but in female subjects, no significant relationships were found after adjustment (beta=0.007, p=0.82). CONCLUSION: These results suggest that GGT is independently associated with an increased level of CRP in both males and females. In addition, in males, GGT is related to an increased level of arterial stiffness. PMID- 17412576 TI - Informativeness of dinucleotide repeat-based primers in fungal pathogen of rice Magnaporthe grisea. AB - Microsatellites or simple sequence repeats (SSRs) occur ubiquitously and show complex patterns in length, motif size and sequence. Among SSRs, dinucleotide repeats occur in high abundance in fungi with shorter length as compared to other organisms. In this study, multilocus profiles obtained in Magnaporthe grisea, a model plant pathogen were evaluated. The results showed lower rate of polymorphism by (GT)(n)/(TG)(n) repeat-based primers and suggested occurrence of (GA)(n)/(AG)(n) repeats as integral repeats and (TC)(n)/(CT)(n) and (AC)(n)/(CA)(n) as non-integral repeats. Low repeat length variation was found to be correlated with less number of repeat motifs. The study provides an insight into the possibility of molecular coevolution of mobile elements and dinucleotide repeats in fungi. The study could be applied to other species for wider applications including evolutionary and population genetics. PMID- 17412577 TI - Differential expression and activity of matrix metalloproteinase-2 and -9 in the calreticulin deficient cells. AB - Calreticulin is an endoplasmic reticulum protein important in cardiovascular development. Deletion of the calreticulin gene leads to defects in the heart and the formation of omphaloceal. These defects could both be due to changes in the extracellular matrix composition. Matrix metalloproteinases (MMP)-2 and MMP-9 are two of the MMPs which are essential for cardiovascular remodelling and development. Here, we tested the hypothesis that the defects observed in the heart and body wall of the calreticulin null embryos are due to alterations in MMP-2 and MMP-9 activity. Our results demonstrate that there is a significant decrease in the MMP-9 and increase in the MMP-2 activity and expression in the calreticulin deficient cells. We also showed that there is a significant increase in the expression level of membrane type-1 matrix metalloproteinase (MT1-MMP). In contrast, there was no change in the tissue inhibitor of matrix metalloproteinase (TIMP)-1 or -2 in the calreticulin deficient cells as compared to the wild type cells. Interestingly, the inhibition of the MEK kinase pathway using PD98059 attenuated the decrease in the MMP-9 mRNA with no effect on the MMP-2 mRNA level in the calreticulin deficient cells. Furthermore, PI3 kinase inhibitor decreased the expression of both the MMP-2 and MMP-9. This study is the first report on the role of calreticulin in regulating MMP activity. PMID- 17412578 TI - Special airway concerns in patients with mucopolysaccharidoses. AB - The mucopolysaccharidoses are comprised of hereditary disorders joined by errant degradation of mucopolysaccharides. The relatively infrequent opportunity to care for these patients is evidenced by a fairly small number of case reports and anecdotal information. Though lifespan is increasing, onset of respiratory pathology or involvement remains portentous. We present two cases that punctuate the need for insightful decision making while managing the airway for these patients. PMID- 17412579 TI - Ascorbic acid supplementation attenuates exercise-induced bronchoconstriction in patients with asthma. AB - BACKGROUND: Previous research has shown that diet can modify the bronchoconstrictor response to exercise in asthmatic subjects. OBJECTIVE: Determine the effect of ascorbic acid supplementation on pulmonary function and several urinary markers of airway inflammation in asthmatic subjects with exercise-induced bronchoconstriction (EIB). METHODS: Eight asthmatic subjects with documented EIB participated in a randomized, placebo controlled double-blind crossover trial. Subjects entered the study on their usual diet and were placed on either 2 weeks of ascorbic acid supplementation (1500 mg/day) or placebo, followed by a 1-week washout period, before crossing over to the alternative diet. Pre- and post-exercise pulmonary function, asthma symptom scores, fraction of exhaled nitric oxide (FENO), and urinary leukotriene (LT) C4-E4 and 9alpha, 11beta-prostagladin (PG) F2] were assessed at the beginning of the trial (usual diet) and at the end of each treatment period. RESULTS: The ascorbic acid diet significantly reduced (p < 0.05) the maximum fall in post-exercise FEV1 (-6.4 +/- 2.4%) compared to usual (-14.3 +/- 1.6%) and placebo diet (-12.9 +/- 2.4%). Asthma symptoms scores significantly improved (p<0.05) on the ascorbic acid diet compared to the placebo and usual diet. Post-exercise FENO, LTC4-E4 and 9alpha, 11beta-PGF2 concentration was significantly lower (p<0.05) on the ascorbic acid diet compared to the placebo and usual diet. CONCLUSION: Ascorbic acid supplementation provides a protective effect against exercise-induced airway narrowing in asthmatic subjects. PMID- 17412580 TI - Flexible and robust strategies for waste management in Sweden. AB - Treatment of solid waste continues to be on the political agenda. Waste disposal issues are often viewed from an environmental perspective, but economic and social aspects also need to be considered when deciding on waste strategies and policy instruments. The aim of this paper is to suggest flexible and robust strategies for waste management in Sweden, and to discuss different policy instruments. Emphasis is on environmental aspects, but social and economic aspects are also considered. The results show that most waste treatment methods have a role to play in a robust and flexible integrated waste management system, and that the waste hierarchy is valid as a rule of thumb from an environmental perspective. A review of social aspects shows that there is a general willingness among people to source separate wastes. A package of policy instruments can include landfill tax, an incineration tax which is differentiated with respect to the content of fossil fuels and a weight based incineration tax, as well as support to the use of biogas and recycled materials. PMID- 17412581 TI - Influence of calcium chloride on the thermal behavior of heavy and alkali metals in sewage sludge incineration. AB - In order to separate and reuse heavy and alkali metals from flue gas during sewage sludge incineration, experiments were carried out in a pilot incinerator. The experimental results show that most of the heavy and alkali metals form condensed phase at temperature above 600 degrees C. With the addition of 5% calcium chloride into sewage sludge, the gas/solid transformation temperature of part of the metals (As, Cu, Mg and Na) is evidently decreased due to the formation of chloride, while calcium chloride seems to have no significant influence on Zn and P. Moreover, the mass fractions of some heavy and alkali metals in the collected fly ash are relatively high. For example, the mass fractions for Pb and Cu in the fly ash collected by the filter are 1.19% and 19.7%, respectively, which are well above those in lead and copper ores. In the case of adding 5% calcium chloride, the heavy and alkali metals can be divided into three groups based on their conversion temperature: Group A that includes Na, Zn, K, Mg and P, which are converted into condensed phase above 600 degrees C; Group B that includes Pb and Cu which solidify when the temperature is above 400 degrees C; and Group C that includes As, whose condensation temperature is as low as 300 degrees C. PMID- 17412582 TI - Microwave inactivation of Escherichia coli in healthcare waste. AB - Public healthcare wastes from the city of Ribeirao Preto, SP, Brazil, pre sterilised in an autoclave, were inoculated with 5 x 10(5) microorganisms of the species Escherichia coli in vegetative form for microwave processing on a laboratory scale. An analysis was made of the influence of radiation exposure time (15, 25, 30 and 40 min) and power per waste mass unit (60, 80 and 100 W/kg) on the percentage of inactivation of the microorganisms at an incoming waste moisture level of 50%. The experimental results were adjusted based on Chick's law. The activation energies and the Arrhenius pre-exponential factors were determined by the least squares method. The kinetic parameters obtained allow one to predict the degree of inactivation achieved with E. coli in typical healthcare waste, based on the radiation exposure time and temperature. For example, the waste disinfection time required for the inactivation level equivalent to 4Log 10 was estimated to range from 48 to 53 min for wastes processed at 100 W/kg and at temperatures of 90-105 degrees C, respectively. Thus, under the operational conditions of the equipment currently used in Ribeirao Preto, the process of inactivation is probably ineffective, since the exposure time to radiation is only 30 min at the average power of approximately 80 W/kg. PMID- 17412583 TI - Synthesis and biological evaluation of diamine-based histamine H3 antagonists with serotonin reuptake inhibitor activity. AB - The synthesis and structure-activity relationships of a series of novel phenoxyphenyl diamine derivatives with affinity for both the histamine H(3) receptor and the serotonin transporter is described. PMID- 17412584 TI - Synthesis and biological activity of 2-alkylbenzimidazoles bearing a N phenylpyrrole moiety as novel angiotensin II AT1 receptor antagonists. AB - A series of 2-alkylbenzimidazoles bearing a N-phenylpyrrole moiety were synthesized and evaluated as a novel class of AT(1) receptor antagonists. Among them, compounds 10a and 10g inhibited [(125)I] AngII-binding affinity to AT(1) receptor at nanomolar level and potently inhibited the Ang II-induced pressor response by oral administration. Moreover, evaluation in spontaneously hypertensive rats showed that 10a is an orally active AT(1) receptor antagonist. PMID- 17412585 TI - Endosymbiont-dependent host reproduction maintains bacterial-fungal mutualism. AB - Bacterial endosymbionts play essential roles for many organisms, and thus specialized mechanisms have evolved during evolution that guarantee the persistence of the symbiosis during or after host reproduction. The rice seedling blight fungus Rhizopus microsporus represents a unique example of a mutualistic life form in which a fungus harbors endobacteria (Burkholderia sp.) for the production of a phytotoxin. Here we report the unexpected observation that in the absence of endosymbionts, the host is not capable of vegetative reproduction. Formation of sporangia and spores is restored only upon reintroduction of endobacteria. To monitor this process, we succeeded in GFP labeling cultured endosymbionts. We also established a laserbeam transformation technique for the first controlled introduction of bacteria into fungi to observe their migration to the tips of the aseptate hyphae. The persistence of this fungal-bacterial mutualism through symbiont-dependent sporulation is intriguing from an evolutionary point of view and implies that the symbiont produces factors that are essential for the fungal life cycle. Reproduction of the host has become totally dependent on endofungal bacteria, which in return provide a highly potent toxin for defending the habitat and accessing nutrients from decaying plants. This scenario clearly highlights the significance for a controlled maintenance of this fungal-bacterial symbiotic relationship. PMID- 17412586 TI - Facial-expression and gaze-selective responses in the monkey amygdala. AB - The social behavior of both human and nonhuman primates relies on specializations for the recognition of individuals, their facial expressions, and their direction of gaze. A broad network of cortical and subcortical structures has been implicated in face processing, yet it is unclear whether co-occurring dimensions of face stimuli, such as expression and direction of gaze, are processed jointly or independently by anatomically and functionally segregated neural structures. Awake macaques were presented with a set of monkey faces displaying aggressive, neutral, and appeasing expressions with head and eyes either averted or directed. BOLD responses to these faces as compared to Fourier-phase-scrambled images revealed widespread activation of the superior temporal sulcus and inferotemporal cortex and included activity in the amygdala. The different dimensions of the face stimuli elicited distinct activation patterns among the amygdaloid nuclei. The basolateral amygdala, including the lateral, basal, and accessory basal nuclei, produced a stronger response for threatening than appeasing expressions. The central nucleus and bed nucleus of the stria terminalis responded more to averted than directed-gaze faces. Independent behavioral measures confirmed that faces with averted gaze were more arousing, suggesting the activity in the central nucleus may be related to attention and arousal. PMID- 17412587 TI - Cooperative begging in banded mongoose pups. AB - Vivid begging displays are common in species with parental care [1, 2]. They are usually seen as the way that rival offspring selfishly compete over parental investment [3], and individuals are expected to respond to the begging of rivals by increasing their own begging intensity [4, 5]. Here I show the opposite - that potential rivals gain direct benefits from begging by littermates, so that begging behavior becomes a collective enterprise, similar to other cooperative activities. I investigate begging in communally breeding banded mongooses (Mungos mungo), where each pup forms an exclusive relationship with a single helper (its "escort"), minimizing competition over food allocation. Escorts were influenced by the total signal emanating from a litter, so that pups who begged at low rates received more food as litter size increased. Focal pups increased their begging when litters were experimentally reduced or littermates were induced to beg at low rates, but they received food at similar rates and showed reduced weight gain - indicating that they were paying a higher cost for a similar reward. These results suggest that offspring can benefit from companions despite conflicts over the allocation of parental investment [6, 7]. Such benefits provide an explanation for observed variation in the expression of parent-offspring conflict. PMID- 17412588 TI - Pericentric chromatin is an elastic component of the mitotic spindle. AB - BACKGROUND: Prior to chromosome segregation, the mitotic spindle bi-orients and aligns sister chromatids along the metaphase plate. During metaphase, spindle length remains constant, which suggests that spindle forces (inward and outward) are balanced. The contribution of microtubule motors, regulators of microtubule dynamics, and cohesin to spindle stability has been previously studied. In this study, we examine the contribution of chromatin structure on kinetochore positioning and spindle-length control. After nucleosome depletion, by either histone H3 or H4 repression, spindle organization was examined by live-cell fluorescence microscopy. RESULTS: Histone repression led to a 2-fold increase in sister-centromere separation and an equal increase in metaphase spindle length. Histone H3 repression does not impair kinetochores, whereas H4 repression disrupts proper kinetochore function. Deletion of outward force generators, kinesins Cin8p and Kip1p, shortens the long spindles observed in histone repressed cells. Oscillatory movements of individual sister chromatid pairs are not altered after histone repression. CONCLUSIONS: The increase in spindle length upon histone repression and restoration of wild-type spindle length by the loss of plus-end-directed motors suggests that during metaphase, centromere separation and spindle length are governed in part by the stretching of pericentric chromatin. Chromatin is an elastic molecule that is stretched in direct opposition to the outward force generators Cin8p and Kip1p. Thus, we assign a new role to chromatin packaging as an integral biophysical component of the mitotic apparatus. PMID- 17412589 TI - Delayed breeding affects lifetime reproductive success differently in male and female green woodhoopoes. AB - In cooperatively breeding species, many individuals only start breeding long after reaching physiological maturity [1], and this delay is expected to reduce lifetime reproductive success (LRS) [1-3]. Although many studies have investigated how nonbreeding helpers might mitigate the assumed cost of delayed breeding (reviewed in [3]), few have directly quantified the cost itself [4, 5] (but see [6, 7]). Moreover, although life-history tradeoffs frequently influence the sexes in profoundly different ways [8, 9], it has been generally assumed that males and females are similarly affected by a delayed start to breeding [7]. Here, we use 24 years of data to investigate the sex-specific cost of delayed breeding in the cooperatively breeding green woodhoopoe (Phoeniculus purpureus) and show that age at first breeding is related to LRS differently in males and females. As is traditionally expected, males that started to breed earlier in life had greater LRS than those that started later. However, females showed the opposite pattern: Those individuals that started to breed later in life actually had greater LRS than those that started earlier. In both sexes, the association between age at first breeding and LRS was driven by differences in breeding career length, rather than per-season productivity. We hypothesize that the high mortality rate of young female breeders, and thus their short breeding careers, is related to a reduced ability to deal with the high physiological costs of reproduction in this species. These results demonstrate the importance of considering sex-specific reproductive costs when estimating the payoffs of life history decisions and bring into question the long-held assumption that delayed breeding is necessarily costly. PMID- 17412591 TI - Uninformative exaggeration of male sexual ornaments in barn swallows. AB - Models of sexual selection suggest that mate-choice preferences are favored because differences between males in their degree of ornamental exaggeration convey useful information about the direct or indirect benefits they have to offer [1-5]. Such arguments assume that variation in male ornament size can be attributed to variation in the degree of sexually selected exaggeration. We provide the first test of this assumption by conducting tail-length experiments in male barn swallows. Over the last twenty years, a large amount of work has shown that female barn swallows are influenced by male tail length when choosing a mate [6-12]. Recent experiments have shown that a combination of natural and sexual selection results in the elongated tail streamer--a tail that is on average across the population about 12 mm (approximately 10%) longer than the aerodynamic optimum [13, 14]. We show that the aerodynamically optimal tail length varies significantly between males, whereas the extent of streamer elongation beyond the optimum does not. Similarly, the aerodynamically optimal tail length significantly predicts observed tail length and conveys information about flight performance, whereas the extent of sexually selected exaggeration of streamer length does not. Therefore, contrary to handicap models of sexual selection, the sexually selected exaggeration of this trait provides females with little information about any aspect of mate quality PMID- 17412590 TI - A cryptic modifier causing transient self-incompatibility in Arabidopsis thaliana. AB - Breakdown of the pollination barrier of self-incompatibility (SI) in older flowers, a phenomenon known as pseudo-self-compatibility or transient SI, has been described as an advantageous reproductive assurance strategy that allows selfing after opportunities for out-crossing have been exhausted [1-9]. Pseudo self-compatibility is quite prevalent as a mixed mating strategy in nature, but the underlying molecular mechanisms are not known. We had previously shown that Arabidopsis thaliana exhibits cryptic natural variation for pseudo-self compatibility, which is uncovered by transformation of different accessions with SI specificity-determining SRK and SCR genes from its self-incompatible sister species A. lyrata[10, 11]. Here, by using this transgenic A. thaliana model, we show that pseudo-self-compatibility is caused by a hypomorphic allele of PUB8, an S-locus-linked gene encoding a previously uncharacterized ARM repeat- and U box containing protein that regulates SRK transcript levels. This is the first gene underlying pseudo-self-compatibility to be identified and the first report in which cryptic natural variation unveiled by a transgene enabled the cloning of a gene for a complex trait. PMID- 17412592 TI - Patient empowerment: does it still occur in the ICU? AB - The theoretical underpinnings of patient empowerment were developed through the work of educators and community psychologists, working primarily with the socially disadvantaged. Empowerment is seen as a philosophy based upon the belief of the inherent worth and creative potential of each individual. Therefore, the aim of this paper is to explore whether this creative potential associated with patient choice that encapsulates empowerment is applicable to the Intensive Care Unit. PMID- 17412593 TI - Engineering the glycosylation of natural products in actinomycetes. AB - Bioactive natural products are frequently glycosylated with saccharide chains of different length, in which the sugars contribute to specific interactions with the biological target. Combinatorial biosynthesis approaches are being used in antibiotic-producing actinomycetes to generate derivatives with novel sugars in their architecture. Recent advances in this area indicate that glycosyltransferases involved in the biosynthesis of natural products have substrate flexibility regarding the sugar donor but also, less frequently, with respect to the aglycon acceptor. Therefore, the possibility exists of altering the glycosylation pattern of natural products, thus enabling an increase in the structural diversity of natural products. PMID- 17412594 TI - Role of insulin-like signalling in Drosophila lifespan. AB - Regulation of lifespan by the insulin/insulin-like growth factor-like signalling (IIS) pathway has been conserved during evolution from the nematode worm to the mouse. In the insect Drosophila, regulation of lifespan by the IIS pathway was established by data showing that many mutations in single genes encoding IIS components result in an increase in lifespan. Recently, however, the focus has shifted from studying the effects of single gene mutations with ubiquitous effects to finding interventions that alter IIS in specific tissues and at specific stages in the life history of the fruitfly, in order to elucidate the signalling pathways at work and the mechanisms by which alterations in the IIS pathway can extend lifespan. PMID- 17412595 TI - A newly discovered post-translational modification--the acetylation of serine and threonine residues. AB - Recent studies on a bacterial virulence factor, YopJ of Yersinia, have led to the realization that the acetylation of serine and threonine residues could be an important form of post-translational modification in eukaryotes. Although the identification of the machinery used for the addition and removal of acetyl groups on serine or threonine residues is in its infancy, the enzymes thus-far studied provide early insight into the mechanism of this newly discovered post translational modification, and hint at its potential importance. For example, acetylation can compete with phosphorylation targeted to the same residues and could, therefore, alter the course of signaling pathways. What are the implications for signal transduction in eukaryotes and how widespread could acetylation of serine and threonine prove to be? PMID- 17412596 TI - Functional amyloid--from bacteria to humans. AB - Amyloid--a fibrillar, cross beta-sheet quaternary structure--was first discovered in the context of human disease and tissue damage, and was thought to always be detrimental to the host. Recent studies have identified amyloid fibers in bacteria, fungi, insects, invertebrates and humans that are functional. For example, human Pmel17 has important roles in the biosynthesis of the pigment melanin, and the factor XII protein of the hemostatic system is activated by amyloid. Functional amyloidogenesis in these systems requires tight regulation to avoid toxicity. A greater understanding of the diverse physiological applications of this fold has the potential to provide a fresh perspective for the treatment of amyloid diseases. PMID- 17412597 TI - Studies of interactions between platinum(II) complexes and some biologically relevant molecules. AB - The reactions of Pt(II) complexes, cis-[Pt(NH3)2Cl2], [Pt(terpy)Cl]+, [Pt(terpy)(S-cys)]2+, and [Pt(terpy)(N7-guo)]2+, where terpy=2,2':6',2'' terpyridine, S-cys=L-cysteine, and N7-guo=guanosine, with some biologically relevant ligands such as guanosine-5'-monophosphate (5'-GMP), L-cysteine, glutathione (GSH) and some strong sulfur-containing nucleophiles such as diethyldithiocarbamate (dedtc), thiosulfate (sts), and thiourea (tu), were studied in aqueous 0.1 M Hepes at pH of 7.4 using UV-vis, stopped-flow spectrophotometry, and 1H NMR spectroscopy. PMID- 17412598 TI - Synthesis and biological evaluation of new 6-s-cis locked 1,2,25 trihydroxyprevitamin D3 analogues. AB - An efficient synthesis of several diastereomers of 2-hydroxy substituted 1alpha,25-dihydroxyprevitamin D3 derivatives was accomplished utilizing a practical route to the A-ring synthon. The biological activity of the analogues was evaluated in vitro. All the synthesized derivatives demonstrated low affinity for the vitamin D receptor and vitamin D-binding protein compared with 1alpha,25 dihydroxyvitamin D3, the natural hormone. 1alpha,2beta,25-trihydroxy-19-nor-pre D3 was the most potent of the analogues in inhibiting proliferation of MCF-7 cells but requires higher EC50 concentrations than 1alpha,25-dihydroxyvitamin D3. PMID- 17412599 TI - Immunogens related to the synthetic tetrasaccharide side chain of the Bacillus anthracis exosporium. AB - The known methyl 2-O-acetyl-3,4-di-O-benzyl-1-thio-alpha-L-rhamnopyranoside (3) was converted to the corresponding 5-methoxycarbonylpentyl glycoside 4 which was deacetylated. The product 5 was used as the initial glycosyl acceptor to construct two trirhamnoside glycosyl acceptors having HO-3(III) flanked by either benzoyl or benzyl groups, compounds 10 and 29, respectively [fully protected, except HO-3(III), alpha-L-Rha-(1-->3)-alpha-L-Rha-(1-->2)-alpha-L-Rha-1-O (CH2)5COOCH3]. When these were glycosylated with ethyl 4-azido-3-O-benzyl-4,6 dideoxy-2-O-bromoacetyl-1-thio-beta-D-glucopyranoside (18), only the benzylated glycosyl acceptor 29 gave good yield of the desired tetrasaccharide 30. The alpha and beta-linked products, together with the corresponding orthoester 23, were formed in almost equal amount when glycosylation of 10 was performed with the glycosyl donor carrying the 2-O-bromoacetyl protecting group. Deprotection at O-2 of 30, followed by further functionalization of the molecule and global deprotection, gave the 5-methoxycarbonylpentyl glycoside of the title tetrasaccharide, beta-Ant-(1-->3)-alpha-L-Rha-(1-->3)-alpha-L-Rha-(1-->2)-alpha-L Rha (35). Except for differences due to presence of the anomeric 5 methoxycarbonylpentyl group, the fully assigned NMR spectra of glycoside 35 were found to be virtually identical to those reported for the parent tetrasaccharide isolated from Bacillus anthracis exosporium, thus proving the correct structure assigned to the naturally occurring substance. All theoretically possible structural fragments of 35, as well as analog of 35 lacking the 2-O-methyl group at the terminal 4,6-dideoxyglucosyl residue, compound 40, were also synthesized. Tetrasaccharide 35, its beta-linked and non-methylated analogs 2 and 40, respectively, as well as the trirhamnoside fragment of 35, glycoside 12, were further functionalized and conjugated to BSA using squaric acid chemistry, to give neoglycoconjugates with a predetermined carbohydrate-protein ratio of approximately 3 and approximately 6. PMID- 17412600 TI - Synthesis and anti-HIV activity of trivalent CD4-mimetic miniproteins. AB - A series of trivalent CD4-mimetic miniproteins was synthesized, in which three CD4M9 miniprotein moieties were tethered on a threefold-symmetric scaffold. The trivalent miniproteins were designed to target the CD4-binding sites displayed in the trimeric gp120 complex of HIV-1. The synthesis takes advantage of the highly efficient ligation between a cysteine-tagged CD4M9 miniprotein and a suitable trivalent maleimide that varied in the nature and length of spacer. Antiviral assay revealed that most of the synthetic trivalent miniproteins demonstrated significantly enhanced anti-HIV activities over the monomeric CD4M9 against both R5- and X4-tropic viruses, indicating the beneficial multivalent effects. One compound that possesses a hydrophobic linker was shown to be 140-fold more active than CD4M9 against HIV-1(Bal) infection, implicating a positive contribution of the lipid portion to the antiviral activity. It was also found that most of the trivalent miniproteins showed comparable anti-HIV activities in comparison with a typical bivalent miniprotein, regardless of the length of the linker. The results implicate a novel mechanism of the interactions between the multivalent inhibitors and the trimeric gp120 complex. PMID- 17412601 TI - Synthesis and antifungal activity of novel s-substituted 6-fluoro-4 alkyl(aryl)thioquinazoline derivatives. AB - 6-Fluoro-4-quinazolinol is prepared by the cyclization reaction of 2-amino-5 fluorobenzoic acid and formamide. The resulting thiol obtained by treatment of hydroxyl group with phosphorus (V) sulfide is converted under phase transfer condition to 4-substituted 4-alkylthio-6-fluoroquinazoline derivatives by reaction with halide. The structures of the compounds are confirmed by elemental analysis, IR, and (1)H NMR. Title compounds 3a, 3g, and 3h are found to possess good antifungal activities. Using the mycelial growth rate method in the laboratory, the mechanism of action of 3g against Fusarium oxysporum in vitro is studied. The results indicate that 3a, 3g, and 3h have high inhibitory effect on the growth of most of the fungi with EC(50) values ranging from 8.3 to 64.2 microg/mL. After treating F. oxysporum with compound 3g at 100 microg/mL, only 6.5% of its spore bourgeoned. The permeability of the cell membrane increases along with the malformation of the hypha and condensation of its endosome. After treatment with compound 3g at 100 microg/mL within 12h, the mycelial reducing sugar, D-GlcNAc, content and chitinase activity decline, but the soluble protein content shows no obvious change. PMID- 17412602 TI - Complement activation in experimental and human temporal lobe epilepsy. AB - We investigated the involvement of the complement cascade during epileptogenesis in a rat model of temporal lobe epilepsy (TLE), and in the chronic epileptic phase in both experimental as well as human TLE. Previous rat gene expression analysis using microarrays indicated prominent activation of the classical complement pathway which peaked at 1 week after SE in CA3 and entorhinal cortex. Increased expression of C1q, C3 and C4 was confirmed in CA3 tissue using quantitative PCR at 1 day, 1 week and 3-4 months after status epilepticus (SE). Upregulation of C1q and C3d protein expression was confirmed mainly to be present in microglia and in a few hippocampal neurons. In human TLE with hippocampal sclerosis, astroglial, microglial and neuronal (5/8 cases) expression of C1q, C3c and C3d was observed particularly within regions where neuronal cell loss occurs. The membrane attack protein complex (C5b-C9) was predominantly detected in activated microglial cells. The persistence of complement activation could contribute to a sustained inflammatory response and could destabilize neuronal networks involved. PMID- 17412603 TI - Effects of gender on nigral gene expression and parkinson disease. AB - To identify gene expression patterns in human dopamine (DA) neurons in the substantia nigra pars compacta (SNc) of male and female control and Parkinson disease (PD) patients, we harvested DA neurons from frozen SNc from 16 subjects (4 male PDs, 4 female PDs, 4 male and 4 female controls) using Laser Capture microdissection and microarrays. We assessed for enrichment of functional categories with a hypergeometric distribution. The data were validated with QPCR. We observed that gender has a pervasive effect on gene expression in DA neurons. Genes upregulated in females relative to males are mainly involved in signal transduction and neuronal maturation, while in males some of the upregulated genes (alpha-synuclein and PINK1) were previously implicated in the pathogenesis of PD. In females with PD we found alterations in genes with protein kinase activity, genes involved in proteolysis and WNT signaling pathway, while in males with PD there were alterations in protein-binding proteins and copper-binding proteins. Our data reveal broad gender-based differences in gene expression in human dopaminergic neurons of SNc that may underlie the predisposition of males to PD. Moreover, we show that gender influences the response to PD, suggesting that the nature of the disease and the response to treatment may be gender dependent. PMID- 17412604 TI - When the sky falls on one's head. PMID- 17412605 TI - Another perspective of the dysphagia due to tuberculosis. PMID- 17412606 TI - Surgical treatment of right-sided active infective endocarditis with or without involvement of the left heart: 20-year single center experience. AB - OBJECTIVE: The aim of this paper is to review the 20-year experience of surgical treatment of right-sided infective endocarditis at our institution, and in particular to compare the outcome of isolated right-sided endocarditis to right sided endocarditis with involvement of the left heart. METHODS: Between April 1986 and April 2006, 84 operations had to be performed in 79 patients (49 men, median age 43.5 years). There were 72 (85.7%) cases of native and 12 (14.3%) of prosthetic valve endocarditis. In 57 (67.9%) cases, operation was for isolated right-sided endocarditis (RSE) and in 27 (32.1%) cases for combined right and left-sided endocarditis (RLSE). Follow-up was completed in all 91% of survivors. The median follow-up time at 448 patient years was 3.59 years (range 6 months 19.27 years). RESULTS: There was a highly significant difference between the survival rates of patients operated on due to RSE compared to RLSE: the 30-day, 1 , 5-, 10- and 20-year survival rate after RSE operation was 96.2%, 88.4%, 73.5%, 70.4% and 70.4%, respectively, compared to 72.0%, 67.8%, 50.8%, 35.6% and 35.6% after operation for RLSE (p=0.0093). Patients with RLSE more often underwent emergency operations (p<0.001), preoperatively were more often on high-dose catecholamines (p<0.006) and intraoperatively showed more abscess formation (p<0.001). Freedom from reoperation at 30 days, 1-, 5-, 10- and 20-year was 97.4%, 95.9%, 92.2%, 88.6% and 88.6%, respectively. Risk factors for early mortality were priority of surgery with an odds ratio (OR) of 5.13, age over 40 years (OR 1.04 and 3.24) and left heart involvement (or 2.54). CONCLUSIONS: Our surgical strategy for right-sided infective endocarditis is based on three principles: (1) debridement of the infected area or vegetectomy; (2) valve repair whenever possible, avoiding artificial material; (3) if valve replacement is unavoidable, use of a biological substitute without any artificial material that might become infected. Following these strategies surgery of right-sided infective endocarditis with or without left-side involvement can be performed with good early, mid-term and long-term results. Patients with involvement of the left side showed not only worse preoperative conditions but also a significantly poorer clinical outcome than those with isolated right-sided infective endocarditis. PMID- 17412607 TI - Paediatric Cushing's syndrome: epidemiology, investigation and therapeutic advances. AB - Cushing's syndrome (CS), which is caused by excessive circulating glucocorticoid concentrations, is rare in the paediatric age range but presents a diagnostic and therapeutic challenge. Most paediatric endocrinologists have limited experience of managing children or adolescents with CS and thus benefit from close consultation with colleagues who treat adult patients. A protocol for investigation is required that broadly follows the model for adult patients. Here, the epidemiology and diagnosis of different causes of CS are discussed according to typical age of presentation. Treatment strategies for adrenocorticotrophic hormone (ACTH)-independent and ACTH-dependent CS are described and critically appraised. The management of paediatric CS patients after cure also presents challenges for optimizing growth, bone health, reproduction and body composition from childhood into and during adult life. PMID- 17412608 TI - Interferon-gamma synergises with tumour necrosis factor and lymphotoxin-alpha to enhance the mRNA and protein expression of adhesion molecules in mouse brain endothelial cells. AB - Changes to the cerebral microvasculature are evident during cerebral malaria (CM). Activation of the endothelium is likely to be due to the actions of cytokines, circulating levels of which are elevated during CM. Endothelial cells are known to up-regulate the expression of cellular adhesion molecules, which can lead to cellular sequestration and obstruction of vessels. However, it is unknown whether cytokines synergise in the up-regulation of the adhesion molecules involved in CM. In this study, the mRNA and/or protein expression of the adhesion molecules vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), P-selectin and E-Selectin were examined in a mouse brain endothelial cell line. Endothelial cells were stimulated with interferon-gamma (IFN-gamma), tumour necrosis factor (TNF) and lymphotoxin-alpha (LT-alpha), alone or in combination. The expression of ICAM-1, VCAM-1, P-selectin and E-Selectin mRNA in mouse brain endothelial cells by TNF and/or LT-alpha was found to be significantly enhanced in the presence of IFN-gamma. The same synergistic effect was found when analyzing ICAM-1 protein expression in cytokine stimulated mouse brain endothelial cells. The findings show that cytokines can synergise to influence gene expression and protein expression in a mouse brain endothelial cell line. PMID- 17412609 TI - Protective effects of amylin on reserpine-induced gastric damage in the rat. AB - Here we show that the vasoactive peptide amylin protects against reserpine induced gastric injury in the rat, resulting in lower score of gastric lesions. Hepatocyte growth factor (HGF), its c-Met receptor and cyclooxygenase-2 (COX-2) expression, usually increased in course of reserpine-induced gastric damage, was decreased in rats treated with amylin. Pretreatment with the specific amylin receptor antagonist AC187 abrogated the gastroprotective effects of amylin and restored high expression levels of HGF, c-Met and COX-2. Our data suggest that protective effects of amylin upon the gastric mucosa are specific and eventually involve modulation of HGF, c-Met and COX-2 expression. PMID- 17412610 TI - Atrophy is associated with posterior cingulate white matter disruption in dementia with Lewy bodies and Alzheimer's disease. AB - Hippocampal atrophy and posterior cingulate hypometabolism are common features of both Alzheimer's disease (AD) and dementia with Lewy bodies (DLB). These regions show correlated activity at rest as part of the 'default network', and they are connected by the cingulum, a white matter (WM) tract. We hypothesised that hippocampal atrophy would be associated with disruption of the cingulum, as determined by diffusion tensor imaging. We recruited 15 people with AD, 16 with DLB, and 15 healthy control subjects of similar age. They were scanned on a 1.5 T MRI system with a T1 weighted 3D sequence and diffusion tensor FLAIR imaging. The T1 images were segmented into grey and white matter and spatially normalised using SPM. Hippocampal atrophy was estimated by calculating the mean grey matter (GM) volume from a region of interest in standard space and global atrophy from the total CSF segmentation. Fractional anisotropy (FA) maps were calculated and also spatially normalised. Using SPM, a multivariate correlation of FA against hippocampal GM, global atrophy and disease group was performed. We found a bilateral region adjacent to the posterior cingulate and encompassing a branch of the cingulum where global atrophy correlated with fractional anisotropy, after controlling for diagnosis and hippocampal GM. The results suggest that dementia disease progression as measured by global atrophy is associated with disruption of the white matter which connects posterior cingulate and lateral parietal regions. Hence, in addition to the hypometabolism in these regions in AD and DLB, there is also disruption to the white matter connecting them. Future studies are needed to determine whether the disruption precedes or is consequent on atrophy or hypometabolism. PMID- 17412611 TI - Neural substrates of driving behaviour. AB - Driving a vehicle is an indispensable daily behaviour for many people, yet we know little about how it is supported by the brain. Given that driving in the real world involves the engagement of many cognitive systems that rapidly change to meet varying environmental demands, identifying its neural basis presents substantial problems. By employing a unique combination of functional magnetic resonance imaging (fMRI), an accurate interactive virtual simulation of a bustling central London (UK) and a retrospective verbal report protocol, we surmounted these difficulties. We identified different events that characterise the driving process on a second by second basis and the brain regions that underlie them. Prepared actions such as starting, turning, reversing and stopping were associated with a common network comprised of premotor, parietal and cerebellar regions. Each prepared action also recruited additional brain areas. We also observed unexpected hazardous events such as swerving and avoiding collisions that were associated with activation of lateral occipital and parietal regions, insula, as well as a more posterior region in the medial premotor cortex than prepared actions. By contrast, planning future actions and monitoring fellow road users were associated with activity in superior parietal, lateral occipital cortices and the cerebellum. The anterior pre-SMA was also recruited during action planning. The right lateral prefrontal cortex was specifically engaged during the processing of road traffic rules. By systematically characterising the brain dynamics underlying naturalistic driving behaviour in a real city, our findings may have implications for how driving competence is considered in the context of neurological damage. PMID- 17412612 TI - Face recognition and cortical responses: effect of stimulus duration. AB - To clarify the relationship between face perception and cortical activation, we manipulated the face recognition performance of 9 subjects by varying the duration (DUR) of stimuli while cortical neuromagnetic responses were recorded. A face image replaced a continuous pixel-noise mask for 17-200 ms, and the subject reported which of the pre-learned faces had been presented. Two cortical responses were clearly stronger to intact than phase-scrambled faces: the temporo occipital response peaking at 140-200 ms (M170) and a more widely distributed response peaking at 200-500 ms (M300). For the shortest DURs (17-33 ms), face recognition was at chance level and the cortical responses negligible. For DURs of 50-83 ms, the proportion of recognized faces as well as the strength of cortical responses increased steeply. Recognition performance saturated at DURs of around 100 ms, whereas cortical responses continued to increase until the longest DUR of 200 ms. Amplitudes of both M170 and M300 were thus tightly correlated with recognition performance (r=0.98), but comparison of the increment rates as a function of DUR showed the recognition performance to have an even closer similarity to M170 than to M300. In single-trial analysis the variability of response strengths increased in a direct proportion to response amplitude, demonstrating the averaged responses to be composed of graded rather than of all or-nothing-type single responses. PMID- 17412613 TI - Old World Shrike-babblers (Pteruthius) belong with New World Vireos (Vireonidae). PMID- 17412614 TI - Phylogeographic patterns, molecular and vocal differentiation, and species limits in Schiffornis turdina (Aves). AB - Establishing species limits can be challenging for organisms in which few variable morphological characters are available, such as Schiffornis turdina, a Neotropical suboscine bird of long-debated taxonomic affinities. Apart from its dull plumage and secretive behavior, this taxon is well-known for its subtle but discrete within-species geographic variation in vocalizations. Phylogeographic reconstruction based on three mitochondrial markers sampled across much of the species' range reveals substantial structuring, concordant with recognized areas of endemism in Neotropical lowland forests. Monophyly of S. turdina was weakly supported by the combined dataset, as was the basal position of the Guyanan Shield population with regard to other S. turdina clades. Based on the results from both genetic and a preliminary, qualitative analysis of vocalizations, I recommend revised species limits to reflect more accurately the evolutionary history of this complex. PMID- 17412615 TI - Epilepsy surgery in tuberous sclerosis: the Dutch experience. AB - INTRODUCTION: Epilepsy associated with tuberous sclerosis complex (TSC) is drug resistant in more than half of the patients. Epilepsy surgery may be an alternative treatment option, if the epileptogenic tuber can be identified reliably and if seizure reduction is not at the expense of cognitive or other functions. We report the pre-surgical identification of the epileptogenic tuber and post-surgical outcome of patients with TSC in The Netherlands. METHODS: Twenty-five patients underwent the pre-surgical evaluation of the Dutch Comprehensive Epilepsy Surgery Programme, including a detailed seizure history, interictal and ictal video EEG registrations, 3D FLAIR MRI scans and neuropsychological testing. Suitability of the candidates was decided in consensus. Seizure outcome, scored with the Engel classification, and cognition were reassessed at fixed post-surgery intervals. RESULTS: Epilepsy surgery was performed in six patients. At follow-up, four patients had Engel classification 1, two had classification 4. Improved development and behaviour was perceived by the parents of two patients. Epilepsy surgery was not performed in 19 patients because seizures were not captured, ictal onset zones could not be localised or were multiple, interictal EEG, video EEG and MEG results were not concordant, or seizure burden had diminished during decision making. A higher cognition index was found in the surgical patients compared to the non-surgical candidates. CONCLUSIONS: Epilepsy surgery can be performed safely and successfully in patients in whom semiology, interictal EEG, ictal EEG, MEG and the location of tubers are concordant. In other cases the risk of surgery should be weighed against the chance of seizure relief and in case of children subsequent impact on neurodevelopment. PMID- 17412616 TI - Delayed post-ictal event-related potentials do not differentiate between generalized tonic-clonic seizures and syncope. AB - BACKGROUND: The differentiation between generalized tonic-clonic seizure (GTCS) and syncope is an important clinical problem. Corroborative investigations, which are requested when history is unclear or insufficient, have limited diagnostic value. The aim of our study was to determine whether auditory event-related potentials (ERPs) can be utilized in post-event differentiation between GTCS and syncope. MATERIALS AND METHODS: ERPs were recorded in 18 patients with a single seizure and in 21 patients following syncope, either on one or two occasions. ERP latencies and amplitudes were compared between groups and sessions. RESULTS: No significant differences of P3, N2, P2 and N1 latencies and P3, N2 and P2 amplitudes were found between patients following a single GTCS as compared to patients following syncope on either session. CONCLUSION: Post-event ERPs are insufficient to differentiate between GTCS and syncope. Further investigations are needed to evaluate the influence of different post-event intervals and ERP paradigms on ERP parameters in patients with GTCS. PMID- 17412617 TI - Neurogenesis and learning: acquisition and asymptotic performance predict how many new cells survive in the hippocampus. AB - Previous research has shown that some associative learning tasks prevent the death of new neurons in the adult hippocampus. However, it is unclear whether it is mere exposure to the training stimuli that rescues neurons or whether successful learning of the task is required for enhanced neuronal survival. If learning is the important variable, then animals that learn better given the same amount of training should retain more of the new cells after learning than animals that do not learn as well. Here, we examined the effects of training versus learning on cell survival in the adult hippocampus. Animals were injected with BrdU to label a population of cells and trained one week later on one of two trace conditioning tasks, one of which depends on the hippocampus and one that does not. Increases in cell number occurred only in animals that acquired the learned response, irrespective of the task. There were significant correlations between acquisition and cell number, as well as between asymptotic performance and cell number. These data support the idea that learning and not simply training increases the survival of the new cells in the hippocampus. PMID- 17412618 TI - Monocyte-derived dendritic cells formed at the infection site control the induction of protective T helper 1 responses against Leishmania. AB - Infection-induced inflammatory reactions involve a strong increase in dendritic cells (DCs) at the infection site and draining lymph nodes (dLNs). Whether inflammatory DCs are recruited to these locations or differentiate locally, and what their functional relevance is, remain unclear. Here we showed that during Leishmania infection, monocytes were recruited to the dermis and differentiated into "dermal monocyte-derived DCs," which subsequently migrated into the dLNs. In addition, monocyte recruitment to the dLNs resulted in the differentiation into "LN monocyte-derived DCs." Analysis of the kinetics of monocyte differentiation into DCs, susceptibility to infection, IL-12 production, and L. major-specific T cell stimulation potential suggest that dermal monocyte-derived DCs controlled the induction of protective T helper 1 responses against Leishmania. Thus, the demonstration of monocyte differentiation potential into DCs during in vivo infection and of local DC differentiation in inflammatory foci suggests that de novo formed monocyte-derived DCs are essential in T cell immunity against pathogens. PMID- 17412619 TI - Clonality and host selection in the wheat pathogenic fungus Puccinia triticina. AB - Clonal reproduction in Puccinia triticina, the cause of wheat leaf rust, has long been hypothesized but has never been demonstrated. Using a population genetics approach and microsatellite markers, we analysed genetic diversity of this fungus at each level of genome organisation. Sampling included isolates from two field populations growing on two cultivars carrying specific resistance genes, completed with isolates representing the main pathotypes identified from a national survey. For the two cultivars, populations differentiated according to the distribution of their genotypes and pathotypes. There was a high proportion of repeated genotypes, combined with a significant linkage disequilibrium and a strong negative value for FIS. These three factors, especially heterozygote excess, strongly support the hypothesis of a high rate of clonal reproduction. Each pathotype matched a unique multilocus genotype, except for a few isolates, which were taken to be mutants of the dominant genotype. We discussed the strong relationship between pathotypes and genotypes as the consequence of clonal reproduction combined with a strong selection exerted by host cultivars. PMID- 17412620 TI - Contrast ultrasound: general principles and veterinary clinical applications. AB - The concept of contrast enhancement has significantly extended the usefulness of ultrasound imaging in human medicine and medical research over the past decade. The persistence and efficacy of ultrasound contrast agents has been improved and specific imaging sequences have been developed. Contrast ultrasound provides Doppler and grey-scale enhancement. Doppler examinations are improved when studying deep or small vessels and vessels with low flow velocities. Specific contrast imaging sequences allow detection of tissue enhancement with grey-scale ultrasound which enables assessment of tissue perfusion. Major clinical applications of contrast ultrasound in the human medicine field are the heart, the parenchymal organs such as the liver, spleen and kidneys, and vascular applications. Many other interesting applications have been identified and beside their diagnostic value, intensive research is currently investigating the use of ultrasound contrast agents for therapeutic applications such as targeted delivery of drug- or gene-loaded microbubbles. In the last few years, contrast ultrasound has also been introduced in veterinary medicine. Its usefulness has been shown in diseases of the liver, spleen, kidney, pancreas, lymph nodes and superficial tumours. In the present article, an overview of the physical principles, imaging techniques and image analyses is presented. In addition, a literature review details the current use in veterinary medicine and areas of potential utilization are discussed. PMID- 17412621 TI - Streptococcus gallolyticus subsp. pasteurianus septicaemia in goslings. AB - Increased mortality was observed in a group of 2-3-week old goslings. At post mortem examination, gross lesions included congestion and haemorrhages in the viscera. Histopathological lesions included generalised congestion of tissues with multifocal bacterial colonies, an acute septic fibrinoid splenitis, and an acute septic hepatitis, all consistent with acute septicaemia. Gram-stained sections of liver and lung confirmed the presence of Gram-positive bacteria and Streptococcus gallolyticus subsp. pasteurianus (Streptococcus bovis biotype II/2) was isolated in pure growth from two of the goslings, and in mixed growth from a third. PMID- 17412622 TI - Temporal changes in serum concentrations of acute phase proteins in newborn dairy calves. AB - Age-dependent changes in blood concentrations of four bovine acute phase proteins (APPs), serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), haptoglobin (Hp) and alpha(1)-acid glycoprotein (AGP), were examined using two groups of newborn dairy calves. APP concentrations were monitored for either 3 weeks (Group A, n=13) or 2 months (Group B, n=13) after birth. Blood was collected at day 0-1 (Group A only), day 3 and then once or twice weekly until the end of the study. The possible transfer of colostral SAA as a source of SAA in the offspring was investigated by determining SAA isoforms in the serum of calves and in colostrum of their dams. Serum concentrations of all four APPs were high after birth, and concentrations showed a gradual decrease during the first 3 weeks of life. The lowest concentrations were at 21 days of age, after which concentrations stabilized. The calves synthesized adult SAA isotypes, and circulating SAA was not derived from colostrum. The results indicated that post partum elevation of APPs is associated with the birth process and/or factors in colostrum and not necessarily with disease-related processes. This stresses the importance of considering a calf's age when interpreting APP concentrations in serum. PMID- 17412623 TI - A new lethal case of joint contractures, facial abnormalities, pachygyria plus early-onset encephalopathy with a suppression-burst EEG pattern. AB - Pachygyria, joint contractures and facial abnormalities have been reported in the literature very rarely. These features constitute a new lethal syndrome. We describe a male infant who represents an additional case of the same lethal syndrome first described in 1989. The patient also presented an early-onset epileptic encephalopathy with a suppression-burst EEG pattern. PMID- 17412624 TI - Water diffusion-exchange effect on the paramagnetic relaxation enhancement in off resonance rotating frame. AB - The off-resonance rotating frame technique based on the spin relaxation properties of off-resonance T1rho can significantly increase the sensitivity of detecting paramagnetic labeling at high magnetic fields by MRI. However, the in vivo detectable dimension for labeled cell clusters/tissues in T1rho-weighted images is limited by the water diffusion-exchange between mesoscopic scale compartments. An experimental investigation of the effect of water diffusion exchange between compartments on the paramagnetic relaxation enhancement of paramagnetic agent compartment is presented for in vitro/in vivo models. In these models, the size of paramagnetic agent compartment is comparable to the mean diffusion displacement of water molecules during the long RF pulses that are used to generate the off-resonance rotating frame. The three main objectives of this study were: (1) to qualitatively correlate the effect of water diffusion-exchange with the RF parameters of the long pulse and the rates of water diffusion, (2) to explore the effect of water diffusion-exchange on the paramagnetic relaxation enhancement in vitro, and (3) to demonstrate the paramagnetic relaxation enhancement in vivo. The in vitro models include the water permeable dialysis tubes or water permeable hollow fibers embedded in cross-linked proteins gels. The MWCO of the dialysis tubes was chosen from 0.1 to 15 kDa to control the water diffusion rate. Thin hollow fibers were chosen to provide sub-millimeter scale compartments for the paramagnetic agents. The in vivo model utilized the rat cerebral vasculatures as a paramagnetic agent compartment, and intravascular agents (Gd-DTPA)30-BSA were administrated into the compartment via bolus injections. Both in vitro and in vivo results demonstrate that the paramagnetic relaxation enhancement is predominant in the T1rho-weighted image in the presence of water diffusion-exchange. The T1rho contrast has substantially higher sensitivity than the conventional T1 contrast in detecting paramagnetic agents, especially at low paramagnetic agent volumetric fractions, low paramagnetic agent concentrations, and low RF amplitudes. Short pulse duration, short pulse recycle delay and efficient paramagnetic relaxation can reduce the influence of water diffusion-exchange on the paramagnetic enhancement. This study paves the way for the design of off-resonance rotating experiments to detect labeled cell clusters/tissue compartments in vivo at a sub-millimeter scale. PMID- 17412625 TI - Memorial symposium in honor of Peter Lutz--Florida Atlantic University. PMID- 17412626 TI - Different fatty acid composition in central and peripheral adipose tissues of the American mink (Mustela vison). AB - Fatty acid (FA) composition in the intraabdominal (IAB), subcutaneous (SC) and peripheral adipose tissues of the semiaquatic American mink (Mustela vison) was examined in comparison to the diet by gas-liquid chromatography. There was a clear compositional gradient from the IAB via SC to peripheral adipose tissues and the anatomically different adipose tissues accumulated or metabolized FA selectively. The total lipids of the body appendages had smaller proportions of saturated (SFA) and larger proportions of monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA) than the lipids of the trunk adipose tissues. Especially n-3 PUFA were enriched in the periphery. The appendages were also characterized with a high ratio of unsaturated FA to SFA, an increased Delta9 desaturation index and increased mean numbers of double bonds and carbon atoms in a FA molecule. The proportions of SFA and MUFA of the diet resembled the trunk adipose tissues while the dietary percentage of n-3 PUFA surpassed those of the trunk fat depots but was lower than those of the peripheral fats. These data confirm that the FA signatures of mammals reflect not only their dietary history but also metabolic modifications of ingested FA. PMID- 17412627 TI - Role of the integrin alphaVbeta3 in mediating increased smooth muscle cell responsiveness to IGF-I in response to hyperglycemic stress. AB - Under usual conditions, the role of IGF-I in vascular cell types is to maintain cellular protein synthesis and cell size, and even excess IGF-I does not stimulate proliferation. In pathophysiologic states, such as hyperglycemia, smooth muscle cells (SMC) dedifferentiate and change their responsiveness to IGF I. During hyperglycemia IGF-I stimulates both SMC migration and proliferation. Our laboratory has investigated the molecular mechanism by which this change is mediated. During hyperglycemia SMC secrete increased concentrations of thrombospondin, vitronectin and osteopontin, ligands for the integrin alphaVbeta3. Activation of alphaVbeta3 stimulates recruitment of a tyrosine phosphatase, SHP-2. Exposure of SMC to IGF-I results in phosphorylation of the transmembrane protein, SHPS-1, which provides a docking site for alphaVbeta3 associated SHP-2. After IGF-I stimulation SHP-2 associates with Src kinase, which associates with the signaling protein Shc. Src phosphorylates Shc, resulting in activation of MAP kinases, which are necessary both for stimulation of cell proliferation and migration. Blocking activation of alphaVbeta3 results in an inability of IGF-I to stimulate Shc phosphorylation. Under conditions of normoglycemia, there are insufficient alphaVbeta3 ligands to recruit SHP-2, and no increase in Shc phosphorylation can be demonstrated in SMC. In contrast, if alphaVbeta3 ligands are added to cells in normal glucose, the signaling events that are necessary for Shc phosphorylation can be reconstituted. Therefore when SMC are exposed to normal glucose they are protected from excessive stimulation of mitogenesis by IGF-I. With hyperglycemia there is a marked increased in alphaVbeta3 ligands and Shc phosphorylation in response to IGF-I is sustained. These findings indicate that in SMC hyperglycemic stress leads to altered IGF-I signaling, which allows the cells to undergo a mitogenic response, and which may contribute to the development of atherosclerosis. PMID- 17412628 TI - Abnormal glucose metabolism in patients treated with antipsychotics. AB - Second-generation (atypical) antipsychotic medications are of great benefit to a wide variety of people with psychiatric disorders, especially patients with schizophrenia. However, one constellation of adverse effects is an increased risk of obesity, diabetes, and metabolic syndrome. Increasing numbers of reports concerning impaired glucose tolerance, diabetes, and ketoacidosis have raised concerns about a possible association between abnormal glucose metabolism and treatment with atypical antipsychotics, although the question is still debated because of the presence of many confounding factors. A close relationship between drug-induced weight gain and risk of diabetes has been reported, emphasizing the role of insulin resistance. However, some cases of diabetes developed independently of weight gain, rather rapidly and possibly progressing to ketoacidosis, thus arguing for a severe impairment of insulin secretion. Another debated question is whether diabetes risk is a class action or a differential action. Although not fully scientifically proven yet, available evidence suggests that clozapine and olanzapine have a higher propensity to induce diabetes and metabolic syndrome compared with other atypical antipsychotic drugs, risperidone and quetiapine. Despite more limited available data, amisulpride, aripiprazole and ziprazidone showed less likelihood of precipitating diabetes. Interestingly, reversibility of drug-related diabetes has been reported with aripiprazole. The choice of atypical antipsychotic medication for a specific patient depends on many factors, but the likelihood of developing diabetes should become an important consideration. When prescribing an atypical antipsychotic, a commitment to careful baseline screening and follow-up monitoring is essential in order to mitigate the risk of developing diabetes and associated complications. PMID- 17412629 TI - Aseptic cure of pulmonary paracoccidioidomycosis can be achieved after a previous subcutaneous immunization of susceptible but not resistant mice. AB - The murine model of paracoccidioidomycosis, the most important South American endemic mycosis, mimics the human disease: resistance is associated with preserved cellular immunity while T-cell anergy is related with susceptibility. In the present study we asked whether a previous s.c. infection which induces strong cellular immunity would protect mice against a lethal pulmonary challenge. It was found that susceptible but not resistant mice developed immunoprotection and aseptic cure of infection. Immunoprotection led to reversal of DTH anergy, increased levels of antibodies and pulmonary IL-12, IL-2 and IL-4 indicating a balanced type 1/type 2 response. On the contrary, no marked differences in A/Sn infection and immunity were observed. Depletion experiments showed that immunoprotection required the cooperative action of CD4(+) and CD8(+) T cells in association with IFN-gamma and IL-12. Altogether, these observations demonstrated that susceptible hosts can develop sterilizing immunity and defined the main immunological requirements to control secondary paracoccidioidomycosis. PMID- 17412630 TI - Obstacles to the development of a safe and effective attenuated pre-erythrocytic stage malaria vaccine. AB - Attenuated pre-erythrocytic stage malaria vaccines protect humans from sporozoite challenge. Current technology limits the feasibility of their commercial scale production and poses a significant risk of contamination with transmissible agents. Overcoming these barriers will be very challenging. In contrast, subunit vaccines appear quite feasible and show great promise as candidate malaria vaccines. PMID- 17412631 TI - Comments on Berthelot et al. review: "Provocative sacroiliac joint maneuvers and sacroiliac joint block are unreliable for diagnosing sacroiliac joint pain". PMID- 17412632 TI - Experimental investigations on ultrasound mediated particle breakage. AB - This paper investigates the effect of high-intensity ultrasound on the breakage characteristics of particles suspended in water. A continuous sonicated flow experimental apparatus is used involving a 24 kHz horn type transducer and continuous in-line particle chord length measurement. The effects of sonication power (150-350 W) and temperature (10-50 degrees C) on the breakage characteristics are investigated. Higher breakage is favored at higher sonication power. An optimum temperature in the range tested is observed to exist between 25 degrees C and 37 degrees C. The acoustic cavitation field is influenced by temperature through a complex interplay of vapor pressure, surface tension and viscosity leading to the optimum observed in particle breakage. The efficiency of ultrasound energy conversion to particle breakage is calculated using calorimetry and found along with the net breakage efficiency to initially increase with temperature followed by a decrease after the optimum. It is found to be independent of input ultrasonic power. The effects of contact time is also investigated. PMID- 17412633 TI - Non-Newtonian models for molecular viscosity and wall shear stress in a 3D reconstructed human left coronary artery. AB - The capabilities and limitations of various molecular viscosity models, in the left coronary arterial tree, were analyzed via: molecular viscosity, local and global non-Newtonian importance factors, wall shear stress (WSS) and wall shear stress gradient (WSSG). The vessel geometry was acquired using geometrically correct 3D intravascular ultrasound (3D IVUS). Seven non-Newtonian molecular viscosity models, plus the Newtonian one, were compared. The WSS distribution yielded a consistent LCA pattern for nearly all non-Newtonian models. High molecular viscosity, low WSS and low WSSG values occurred at the outer walls of the major bifurcation in proximal LCA regions. The Newtonian blood flow was found to be a good approximation at mid- and high-strain rates. The non-Newtonian Power Law, Generalized Power Law, Carreau and Casson and Modified Cross blood viscosity models gave comparable molecular viscosity, WSS and WSSG values. The Power Law and Walburn-Schneck models over-estimated the non-Newtonian global importance factor I(G) and under-estimated the area averaged WSS and WSSG values. The non Newtonian Power Law and the Generalized Power Law blood viscosity models were found to approximate the molecular viscosity and WSS calculations in a more satisfactory way. PMID- 17412634 TI - Histone deacetylase inhibitors: signalling towards p21cip1/waf1. AB - Chromatin-modifying enzymes such as histone deacetylases (HDAC) facilitate a closed chromatin structure and hence transcriptional repression. HDAC are commonly affected in human cancer diseases. Thus, inhibition of HDAC represents a novel therapeutic approach. Several studies have shown that HDAC inhibitors strongly activate the expression of the cyclin-dependent kinase inhibitor p21(cip1/waf1) through (i) enhanced histone acetylation around the p21(cip1/waf1) promoter and (ii) the Sp1 sites on the p21(cip1/waf1) promoter releasing the repressor HDAC1 from its binding. p21(cip1/waf1) expression is regulated in a p53 dependent and p53-independent manner. The decision if p21(cip1/waf1) up regulation results in cell cycle arrest or apoptosis, decides about the therapeutic efficacy of an anti-cancer treatment with HDAC inhibitors. PMID- 17412635 TI - Plant development: introducing trehalose metabolism. AB - Trehalose metabolism, a short side-branch of primary carbon metabolism that is controlled by a surprisingly large gene family, is emerging as an important new regulatory pathway in plants. Two recent studies by Namiko Satoh-Nagasawa et al. and Leonardo Gomez et al. have highlighted its novel and possibly pivotal role in coordinating carbon supply with plant growth and development. PMID- 17412636 TI - A genotypic and phenotypic comparison of type III secretion profiles of Pseudomonas aeruginosa cystic fibrosis and bacteremia isolates. AB - The type III secretion system (TTSS) of Pseudomonas aeruginosa enables delivery of a number of toxins involved in the disruption of eukaryotic epithelial surfaces. Whilst the ability to secrete ExoS facilitates invasion and internalization, the secretion of ExoU mediates acute cytotoxicity. In order to determine any association with the ability to secrete these toxins with the nature and severity of human infection, the TTSS genotypes and phenotypes of 163 clinical isolates were determined by multiplex PCR and Western blotting. An exoS+/exoU- genotype was associated with chronic infection in patients with cystic fibrosis whilst an exoS-/exoU+ genotype was associated with strains isolated from blood. Secretion of the ExoU protein was more commonly seen in isolates obtained from blood, suggesting this ability may be important in the development of acute invasive infection. Detection of TTSS toxins in clinical material may be useful in targeting antimicrobial therapy or identifying individuals infected with aggressive strains of P. aeruginosa. PMID- 17412637 TI - Molecular analysis of the thymidine-auxotrophic small colony variant phenotype of Staphylococcus aureus. AB - Thymidine-auxotrophic small colony variants (SCVs) of Staphylococcus aureus are frequently isolated from the chronically infected airways of patients suffering from cystic fibrosis. To date, little is known regarding the molecular mechanisms leading to the formation of this special phenotype, but the auxotrophism for thymidine suggests that impaired thymidine metabolism might play a major role. Sequence analysis of the thymidylate synthase-encoding thyA gene of six clinical thymidine-auxotrophic S. aureus SCVs revealed that all isolates had mutations within thyA. In five isolates the function of the thymidylate synthase was definitely impaired: three of them showed a truncation of the thyA coding sequence by nonsense or frame-shift mutations, in one further isolate the active site of the enzyme was affected by an internal 12-bp deletion, and another isolate had a 173-bp deletion spanning the 5'-terminal region of thyA and the preceding DNA sequence. The sixth isolate showed two amino acid substitutions within the thyA gene product. To confirm the importance of impaired thymidylate synthase synthesis or activity for the formation of the thymidine-auxotrophic SCV phenotype, we constructed a thyA knock-out mutant of a wild-type S. aureus strain. This mutant showed all characteristics of clinical SCVs, such as slow growth, decreased pigment production, reduced hemolytic activity, auxotrophism for thymidine, resistance to trimethoprim/sulfamethoxazol, and reduced plasma coagulase activity. Complementation of the thyA knock-out mutant with intact thyA in trans nearly restored the normal phenotype. In conclusion, these data confirm at the molecular level that impaired thymidylate synthase function is causative for the formation of the thymidine-auxotrophic SCV phenotype in S. aureus. PMID- 17412638 TI - Case-case study of a Salmonella outbreak: an epidemiologic method to analyse surveillance data. AB - Routine infectious disease surveillance data has to be analysed quickly in order to prevent further disease transmission. This can be done by using epidemiological studies, such as case-control studies. However, these studies are complex to perform and susceptible for biases. An alternative is the case-case study design which is less prone to such limitations. So far mainly methodological publications of this study design exist. In this investigation, outbreak cases are compared with cases infected with the same disease. A Salmonella outbreak was analysed with a case-case study design in order to test the applicability of this method. The analysis showed significant associations to the consumption beef (OR 11.8; 95% CI 2.4-66.7; p<0.001) and pork (OR 8.44; 95% CI 1.7-46.4; p<0.001). A case-control study on the same outbreak confirmed these results. An infected control group is very comparable to the outbreak cluster, which limits selection and recall bias. However, the calculated OR has to be interpreted in context with the study design. The case-case study design has proven to be a useful tool to analyse surveillance data. It was easy to perform and had methodological benefits. PMID- 17412639 TI - Influence of industrial sources on children's health--hot spot studies in North Rhine Westphalia, Germany. AB - The aim of this study was to evaluate exposure and health outcome of children living close to industrial sources. Exposure and health outcome was assessed in nearly 1000 children at school beginner age living in the vicinity of industrial sources of three different hot spots (Duisburg North, Duisburg South and Dortmund Horde) and in a rural area of North Rhine Westphalia (NRW), Germany. The cross sectional study was undertaken between March and May 2000. Exposure assessment comprised modelling of ambient air quality data and human biomonitoring (HBM). Depending on the site-specific contaminants, HBM included the measurement of PAH (polycyclic aromatic hydrocarbons) and benzene metabolites in urine as well as heavy metals in blood and urine. Markers of early effects were DNA strand breaks as measured by the comet assay in lymphocytes and excretion of alpha-1 microglobuline and N-acetyl-beta-D-glucosamidase in urine. Health outcome was assessed by questionnaire, lung function test, dermatological examination as well as by RAST (radioallergosorbent test), patch tests and prick tests. The influence of exposure variables on biomarkers and health outcome was measured by means of multiple linear and logistic regression analysis. The most striking results were as follows. Children living close to a coke oven plant (Duisburg North) had increased levels of PAH metabolites in urine, and DNA exposure was increased. Children living at the Dortmund Horde hot spot (increased chromium and nickel ambient air levels from a steel mill) revealed a high prevalence of allergic sensitizations. Sensitization, especially against nickel, was associated with the current internal nickel exposure, and nickel in ambient air was positively associated with the frequency of allergic symptoms. Children from the hot spot areas had increased specific airway resistance and total lung capacity as compared to those of the reference area. In Duisburg North particularly, specific airway resistance and total lung capacity significantly increased with increasing TSP (total suspended particulate). The only positive associations between external and internal exposure were found between benzo[a]pyrene in ambient air and 1-hydroxypyrene in urine, and between lead in ambient air and in the blood of the children. It is concluded that despite improvements of the general air quality during the last decades, living in the vicinity of industrial sources results to some extent in increased internal contaminant exposure and in effects on health outcome. Still ongoing studies are aimed to find out whether the increased PAH and DNA exposure of children from Duisburg North had decreased after the coke oven plant had been shut down in 2003, and if the striking results on the high prevalence of allergic sensitization can be confirmed by introducing an expanded cross-sectional study at four hot spots with increased chromium and nickel ambient air levels. PMID- 17412640 TI - Azygos vein aneurysm--a case for elective resection. AB - Azygos vein aneurysm is a rare differential diagnosis for a posterior mediastinal mass lesion. It is often an incidental finding in a routine chest radiograph. Most azygos vein aneurysms are secondary to conditions causing elevated flow or pressure in the azygos system, like cardiac failure or cirrhosis of the liver. In this group of patients, once diagnosis is established, no treatment is required. Even in the idiopathic azygos vein aneurysms, where no primary cause can be identified, the recommendation so far has been to continue to observe rather than elective excision, since complications are unknown. We present the report of a 73 year old man who underwent excision of an azygos vein aneurysm, which contained a large, free floating thrombus. There has been another reported case of this complication as well [Gomez M-A, Delhommais A, Presicci PF, Besson M, Roger R, Alison D. Partial thrombosis of an idiopathic azygos aneurysm. Br J Radiol 2004;77:342-43].(1) In view of this we feel that it is safer to resect an idiopathic azygos vein aneurysm electively to prevent the possibility of pulmonary embolism. PMID- 17412641 TI - Early warning systems for malaria in Africa: from blueprint to practice. AB - Although the development of early warning systems for malaria has been advocated by international agencies and academic researchers for many years, practical progress in this area has been relatively modest. In two recent articles, Thomson et al. provide new evidence that models of malaria incidence that incorporate monitored or predicted climate can provide early warnings of epidemics one to five months in advance in semi-arid areas. Although the potential benefits of these models in terms of improved management of epidemics are clear, several technical and practical hurdles still need to be overcome before the models can be widely integrated into routine malaria-control strategies. PMID- 17412642 TI - Unexpected rupture of the left ventricular free wall in the echo-lab. AB - BACKGROUND: Left ventricular free wall rupture is an uncommon but catastrophic event following myocardial infarction, and considered the second leading cause of death in acute myocardial infarct. Different types of rupture exist from acute to sub acute types, but prognosis is usually poor. Early recognition and aggressive treatment is recommended. CASE REPORT: We present a case of a 75-year-old man who was referred to our echo-lab for an out patient evaluation because of 1-week duration of worsening of chest pain. Standard transthoracic echocardiography showed hypokinesia in the apical portion of the anterior wall and basal portion of the inferior wall. The patient complained of shortness of breath immediately after the conclusion of the exam, and soon afterward became unconscious. Renewed echocardiography approximately 1 min after syncope displayed a newly developed echo-lucent rim around the heart consistent with left ventricular free wall rupture. Resuscitation was performed followed by attempts to evacuate the blood by needle aspiration, which failed. Open pericardiocentesis stabilised the patient until surgery could be performed. The patient survived and could be discharged 2 weeks later. CONCLUSION: This case highlights the fact that rapid and accurate diagnosis is essential if patients with left ventricular free wall rupture are to survive. PMID- 17412643 TI - Severe calcification of the lateral mitral annulus in constrictive pericarditis: a potential pitfall for the use of echocardiographic tissue Doppler imaging. AB - According to the guidelines of the European Society of Cardiology on the diagnosis and management of pericardial diseases, tissue Doppler imaging (TDI) is proposed to be part of the diagnostic work-up in patients with suspected constrictive pericarditis (CP). We describe a case which illustrates that TDI analysis may be misleading in patients with severe pericardial calcifications of the lateral mitral annulus. Multi-slice computed tomography (MSCT) data in this case contributed much to a better understanding of the impact of heterogeneous calcification patterns on the results of TDI assessment in CP. PMID- 17412645 TI - Different clinical manifestations of hyperammonemic encephalopathy. AB - Valproate is an effective anticonvulsant. Although it is usually well tolerated, it has been associated with many neurological, hematopoietic, hepatic, and digestive system side effects. Among these side effects, hyperammonemia without clinical or laboratory evidence of hepatotoxicity is rare and is an important clinical consideration. The aim of this article was to evaluate the reasons for the unexpected symptoms observed in seven patients with epilepsy patients during valproate treatment. We evaluated seven adult patients with localization-related epilepsy who presented with different acute or subacute neurological symptoms related to valproate-induced hyperammonemic encephalopathy. Four of the seven patients had acute onset of confusion, decline in cognitive abilities, and ataxia. Two had subacute clinical symptoms, and the other patient had symptoms similar to those of acute toxicity. These unusual clinical symptoms and similar cases had not been reported in the literature before. Serum ammonia levels were elevated in all seven patients. After discontinuation of valproate, complete clinical improvement was observed within 5-10 days. On the basis of our work, we suggest that the ammonia levels of a patient who has new neurological symptoms and has been taking valproate must be checked. Clinicians should be aware that these clinical symptoms may be related to valproate-induced hyperammonemic encephalopathy. The symptoms have been observed to resolve dramatically after withdrawal of the drug. PMID- 17412646 TI - Endocrine modulating actions of a phytosterol mixture and its oxidation products in zebrafish (Danio rerio). AB - In this study, a phytosterol preparation ("ultrasitosterol"; 80% beta-sitosterol) and an oxidized ultrasitosterol preparation were evaluated for reproductive effects in zebrafish. Adults were exposed in a continuous flow to 10 microg/L and 100 microg/L ultrasitosterol and oxidized ultrasitosterol, and to 0.27 microg/L 17beta-estradiol and 0.28 microg/L testosterone for 3 weeks. Biomarkers analysed included plasma vitellogenin, testosterone, 11-ketotestosterone, 17beta estradiol, and gonadal histopathology. Ovarian steroid production of testosterone and 17beta-estradiol was examined in isolated zebrafish follicles exposed in vitro to the compounds at the same concentrations as in vivo. Vtg was induced in males exposed to ultrasitosterol, and in males and females exposed to 17beta estradiol. Males exposed to oxidized phytosterols showed increased levels of testosterone and 11-ketotestosterone, and accelerated spermatogenesis. Increased follicular atresia was observed in females exposed to oxidized phytosterols and 17beta-estradiol. Correlation analyses between biomarkers revealed more intercorrelated values for females than for males, and the strongest associations were found in females exposed to oxidized phytosterols. Testosterone production was significantly increased in follicles exposed to the oxidized phytosterol preparations. These findings indicate that the phytosterol mixture is weakly estrogenic in male fish, and that the oxidized phytosterol mixture contains substances that may interfere with spermatogenesis, oogenesis and gonadal steroidogenesis in zebrafish. PMID- 17412647 TI - An adaptive Simon Two-Stage Design for Phase 2 studies of targeted therapies. AB - The field of specialized medicine and clinical development programs for targeted cancer therapies are rapidly expanding. The proposed Phase 2 design allows for preliminary determination of efficacy that may be restricted to a particular sub population defined by biomarker status (presence/absence). The design is an adaptation of the Simon Two-Stage Design. We provide examples where the adaptation can result in substantial savings in sample size and thus potentially lead to greater efficiency in decision making during the drug development process. PMID- 17412648 TI - Challenges and opportunities in processing muscle sympathetic nerve activity with wavelet denoising techniques: detecting single action potentials in multiunit sympathetic nerve recordings in humans. AB - An important issue in analysis of muscle sympathetic nerve activity (MSNA), particularly those measures made in humans, is the problem that background noise of varying levels from recording to recording may interfere with accurate assessment of neural discharge patterns and overall activity. In this study, the utility of wavelet denoising approaches for processing MSNA signals was examined with emphasis on 1) determining whether this approach could improve the signal-to noise (SNR) in the integrated neurogram, and 2) detecting intra-burst single action potential spikes. The utility of wavelet denoising was examined in simulated data and in original human data with three recordings of varying SNR (low, moderate and high) obtained from two healthy individuals. Only in the high SNR signal was the noise removed without concurrent loss of signal. Using a threshold-detecting algorithm individual depolarization spikes were detected in denoised recordings of high original SNR (>3:1) from four individuals and the interspike interval characteristics of these were quantified on a burst-by-burst basis. Compared with baseline (15+/-1 spikes/burst) a reflexive increase in spike count (29+/-4 spikes/burst) was observed during a held maximal inspiration (P<0.01) with concurrent reductions in inter-spike interval (P<0.01). The findings indicate that within multiunit bursts of sympathetic neural activity in the band-pass filtered neural signal, there are particular frequency components that appear to be shared between the signal and noise. This may limit the utility of wavelet denoising to enhance detection of neural bursts in the integrated neurogram of MSNA. However, opportunities exist with this approach to detect variations in action potential contributions within each burst of MSNA. This latter observation suggests that this denoising approach provides a new probe to explore MSNA discharge patterns. PMID- 17412649 TI - Arrested replication fork processing: interplay between checkpoints and recombination. AB - The arrest of DNA replication by DNA damage, nucleotide depletion, DNA-protein complexes or following clashes between transcription and replication factors all have the capacity to promote genome instability. In this review, we discuss how DNA replication is regulated by the checkpoint pathways that stabilise arrested replication forks and the recombination factors that process specific DNA structures resulting from fork arrest. We examine what is known about the interplay between the checkpoints and the recombination apparatus and review the evidence for a recombination-based fork restart pathway in eukaryotes. PMID- 17412650 TI - Ischemic preconditioning induces XRCC1, DNA polymerase-beta, and DNA ligase III and correlates with enhanced base excision repair. AB - Neuronal protection induced by ischemic preconditioning has an important role in the reduction of stroke volume and attenuation of neuronal cell death. Ischemic injury is associated with increased oxidative DNA damage, and failure to efficiently repair these oxidatively damaged lesions results in the accumulation of mutations and neuronal cell death. Although the effects of ischemic tolerance can have profound implications, the precise mechanisms mediating this phenomenon remain unclear. The base excision repair (BER) pathway has a major role in the repair of oxidative DNA base damage after ischemic injury. Using a rat model of ischemic preconditioning, we now report that the neuronal protection observed after induction of ischemic tolerance is associated with increased BER. In situ detection of single-strand breaks and apurinic/apyrimidinic sites reduced to baseline levels after reperfusion following ischemic preconditioning. By contrast, no change was seen in the quantity of in situ lesions after reperfusion in non-ischemic preconditioned brain. Induction of the BER proteins XRCC1, DNA polymerase-beta, and DNA ligase III was seen after reperfusion in ischemically conditioned brain. Moreover, an increase in binding between XRCC1 and DNA polymerase-beta was seen under these conditions, as might be expected during formation of functional BER complexes. Using in vitro BER oligonucleotides, we directly demonstrated an increase in total BER capacity of nuclear extracts prepared from ischemic-conditioned brain after reperfusion compared with sham operated brain. These findings provide direct evidence that increased BER is associated with the neuroprotection induced after ischemic preconditioning, and provides important new mechanistic insight into the important biologic pathways that protect neurons against irreversible ischemic injury. PMID- 17412651 TI - Hypomagnesaemia in cystic fibrosis patients referred for lung transplant assessment. AB - BACKGROUND: Hypomagnesaemia in patients with cystic fibrosis (CF) is under recognized although the true incidence is unknown. Many patients are asymptomatic, although severe deficiency may be associated with muscle weakness, cramps and tetany. Hypomagnesaemia may be a risk factor for post-transplant complications including convulsions, which may be exacerbated by the use of calcineurin inhibitors. The aims of the present study were to describe serum magnesium levels and to investigate the relationship between magnesium levels and age, and renal function measurements in patients with CF referred to a transplant centre for lung transplant assessment. METHODS: We reviewed the data of all 106 CF patients referred for transplant assessment from January 1995 to December 2003. Demographic and biochemical data were recorded and the explanatory variables were subjected to univariate analysis and linear regression analysis. RESULTS: Mean serum magnesium level was 0.75 mmol/L (range 0.46-1.03, normal range 0.74-1.1). 57% of patients had hypomagnesaemia. Serum magnesium levels were not associated with age, serum creatinine or GFR. CONCLUSIONS: Hypomagnesaemia is a common finding in patients with CF referred for lung transplant assessment. Serum magnesium levels should be monitored in all CF patients being referred for lung transplant irrespective of the results of other renal function tests. PMID- 17412652 TI - Cough reflex sensitivity is decreased in female obese patients with obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is associated with impaired airway reflexes. Cough is the main airway defense mechanism but the effect of OSA on cough is unknown. Thirty-two female obese patients scheduled to undergo bariatric surgery were studied. They were classified as presenting OSA (20 patients) when the apnea hypopnea index (AHI) was greater than 5h. Cough sensitivity was measured with citric acid. Increasing concentrations of nebulized citric acid were delivered until cough was elicited. The concentrations eliciting one (C1) and two coughs (C2) were recorded and log transformed (log C1 and log C2). log C1 and log C2 (median (interquartile)) were 1.90 mg/mL (0.90) and 2.2mg/mL (0.30) in OSA patients and 1.60 mg/mL (0.45) and 1.60 mg/mL (0.45) in non-OSA patients, respectively (comparison between groups: p=0.0372 for log C1 and p=0.0227 for log C2). A significant relationship was observed between AHI and log C1 and log C2. Cough sensitivity is therefore, decreased in female obese OSA patients and this decreased sensitivity is positively correlated with disease severity. PMID- 17412653 TI - Long-term facilitation (LTF) and obstructive sleep apnea. PMID- 17412654 TI - Commentary on: "Major differences in the pulmonary circulation of birds and mammals" by John B. West, Rebecca R. Watson and Zhenxing Fu. PMID- 17412655 TI - The evolution of adult height in Europe: a brief note. AB - This paper presents new evidence on the evolution of adult height in 10 European countries for cohorts born between 1950 and 1980 using the European Community Household Panel (ECHP), which collects height data from Austria, Belgium, Denmark, Finland, Greece, Ireland, Italy, Portugal, Spain, and Sweden. Our findings show a gradual increase in adult height across all countries. However, countries from Southern Europe (Greece, Italy, Portugal, and Spain) experienced greater gains in stature than those located in Northern Europe (Austria, Belgium, Denmark, Finland, Ireland, and Sweden). PMID- 17412656 TI - Influence of a novel radiopacifier on the properties of an injectable calcium phosphate cement. AB - An injectable calcium phosphate cement (CPC) with excellent radiopacity was proposed by introducing a novel radiopacifier, strontium carbonate, into the powder phase of CPC. The results showed that the cement showed improved radiopacity even when the content of strontium carbonate was only 8 or 12wt.%. The addition of 8 or 12wt.% strontium carbonate clearly improved the injectability and compressive strength of the cement. Furthermore, the addition of strontium carbonate influenced the pore distribution in the cement. An injectable CPC containing 8 or 12wt.% strontium carbonate has the potential for use in procedures such as vertebroplasty and kyphoplasty. PMID- 17412657 TI - Evaluation of intermolecular interactions of self-etch dentin adhesive primer molecules with type 1 collagen: computer modeling and in vitro binding analysis. AB - The objective of this investigation was to study adhesion of self-etch primer systems to dentin by computer-modeled docking simulations and in vitro binding assay methods. Computer modeling employed analysis of docking simulations of a self-etch primer molecule 10-methacryloxydecamethylene phosphoric acid (MDP) and its calcium salt (MDPCa) as ligands. Typical type 1 collagen segments were selected as targets to reflect potential differences in the amino acid residues in dentinal type 1 collagen triple helix motif. The binding assay involved immunochemical analysis of the modification of anti-collagen binding to collagen by prior exposure of the demineralized dentin to MDP. The estimated mean docking energy values ranged between -4.5 and -8.9kcalmol(-1). The results revealed significant differences in the docking energy estimates as a function of ligand and target structures (p<0.01). Van der Waals and electrostatic contributions were also significantly influenced by ligand selection and collagen structure. Both MDP and MDPCa appear to be important in the overall interactions. Binding assay studies also lend evidence of collagen-ligand intermolecular interactions. It is suggested that the ability of self-etch dentin primer systems to bond effectively to dentin is not limited to the interaction of the primer with the hydroxyapatite of dentin, but also due to the ability to prime dentin efficiently through intermolecular interactions between the primer and its calcium salt with the collagen matrix. Virtual screening methods may be very valuable to select primer molecules for dentin bonding. PMID- 17412659 TI - The "lively" cytokines network in beta-Thalassemia Major-related osteoporosis. AB - Osteoporosis affects approximately 40-50% of adult patients with beta-Thalassemia Major (beta TM). Recent data have implicated an altered modulation of the osteoprotegerin (OPG)/receptor activator of NFkB ligand (RANKL) system in the pathogenesis of beta TM-osteoporosis. OPG/RANKL system acts downstream from IL-1 alpha, IL-6 and TNF-alpha and it may be the final actor mediating the effects of these cytokines on the regulation of both postmenopausal and metabolic bone resorption. However, to date, there are no data on circulating levels of these pro-resorptive cytokines in beta TM patients. We investigated the potential relationships among these cytokines, several markers of bone turnover and bone mineral density (BMD) in beta TM patients. IL-1 alpha, IL-6 and TNF-alpha, OPG and RANKL serum levels, hemato-urinary bone remodeling markers and bone mineral density (BMD) at L2L4 and femoral neck as well as erythropoietin (EPO), 17beta estradiol, and free-testosterone levels were measured in 30 well treated beta TM patients and in 20 healthy subjects, matched for age, sex and BMI with the patients. beta TM patients showed an altered bone turnover, with increased deoxypyridinoline (D-PYR) levels (P<0.0001), decreased osteocalcin (BGP) concentrations (<0.0001) and significantly lower lumbar (P=0.001) and femoral (P<0.05) BMD values as compared to controls. Circulating levels of IL-1 alpha (P<0.0001), TNF-alpha (P<0.0001) and IL-6 (P<0.05) were all increased in beta TM patients as compared with controls. In beta TM patients, IL-1 alpha was significantly related with D-PYR (r=0.5; P<0.05), RANKL (r=0.7; P=0.03) and IL-6 (r=0.3; P=0.006); IL-6 was also significantly correlated with D-PYR (r=0.5; P<0.05) and EPO levels (r=0.3; P=0.03); TNF-alpha showed a negative correlation with L2L4 BMD (r=-0.4; P<0.05). Our data demonstrate, for the first time, an association between increased circulating levels of pro-resorptive cytokines and an altered bone turnover in beta TM-patients, suggesting their involvement in the pathogenesis of beta TM-osteoporosis. PMID- 17412660 TI - [Consciousness experience]. PMID- 17412661 TI - [Delirium]. AB - Delirium is very common in the elderly. It complicates both psychiatric and somatic disorders and is associated with reduced survival, poor functional results, increased duration of hospital stay, and institutionalization. Diagnosis remains difficult in spite of the improvement of the diagnostic criteria, due to the polymorphism of the clinical signs and fluctuation of vigilance and cognition. Age over 70 and previous cognitive impairment are the main risk factors. Precipitating factors are medical and surgical pathologies, intoxications, especially by therapeutic drugs. Delirium can reveal or complicate a previous dementia. Prevention of delirium and care of the delirious patient require the participation of both the medical and nursing staff. PMID- 17412662 TI - [Cognitive aging considered from the point of view of cognitive neurosciences of consciousness]. AB - Linking together cognitive neurosciences of aging with the study of psychological and neurophysiological properties of consciousness opens rich fundamental and clinical perspectives. Indeed, several aspects of cognitive aging seem to deal with modifications of processes directly involved in conscious processing. In the light of this relation between consciousness and aging, three tracks of experimental research are proposed and discussed. PMID- 17412663 TI - [Bipolar disorders and dementia: fortuitous association or filiation? A case report and review of the literature]. AB - The prevalence of bipolar disorders is about 4% of the population. Among the patients the emergence of cognitive disorders has been identified. These disorders can sometimes result in authentic dementia-like syndromes. The neuropsychological characteristics of the cognitive impairment include disorders of the steady attention, executive functions, memory and language. This group of cognitive disorders leads to a subcorticofrontal syndrome. Different neuroanatomical bases have been suggested. The most common etiopathogenic hypothesis retains a cumulative neurological toxicity of the thymic episodes (depression or mania). The role of psychoactive treatments must be taken into account. The secondary cognitive effects of psychoactive drugs usually used in bipolar disorder (lithium, antiepileptics, antidepressants, antipsychotics or tranquillizers) are similar to the cognitive impairment associated with the disease. Differentiation between cognitive disorders related to the pathology and those related to its treatment is difficult. The optimal use of the therapeutic drug classes, preferring antipsychotics of second generation and avoiding products with anticholinergic effect, is essential. There is no specific pharmacological treatment for this dementia syndrome. Psychological approaches offer an alternative way of care, which remains to be investigated and validated. Prevention of iatrogenic effects of drugs constitutes, presently, the main therapeutic intervention. PMID- 17412664 TI - [Cerebral amyloid angiopathy]. AB - Cerebral amyloid angiopathy is characterised by the presence of protein deposits in the wall of the cerebral vessels. Sporadic forms with deposits of Abeta peptide are the most frequent. The diagnosis of cerebral amyloid angiopathy is only certain when amyloid deposits can be visualized on cerebral tissue by biopsy, that appears in most circumstances to carry too many risks for the expected benefits, or post mortem. The clinical diagnostic criteria are advanced age, morphology of the intracerebral hematomas, occurrence of several episodes of intracerebral haemorrhages and presence of dementia. Recent progresses in the field of MRI make the diagnosis of cerebral amyloid angiopathy easier and more accurate. MRI should be more frequently performed in aged patients. PMID- 17412665 TI - [Text comprehension, cognitive resources and aging]. AB - Aging brings cognitive changes. Language is not immune to these changes. The use of compensation strategies may permit older adults to achieve a performance level identical to the one obtained by younger adults. This research aims to study text comprehension in aging and the reading strategies used for by older and younger adults. Kintsch's cognitive model (1988) allows the identification of different levels of representation within text treatment (linguistic form, macrostructure, microstructure and situation model) and predicts the underlying cognitive components. Eye-tracking analyses during reading permit inference about the moments of reading treatment and detection of reading strategies. Sixty highly educated participants were assessed. They were divided in two age groups (20-40 and 60-80 years old). Participants were asked to read and understand three texts constructed to highlight the features of text comprehension within each one of the different levels of text representation. The amount of detail and the necessity of updating the situation model varied for each text. Eye movements were registered by an eye-tracker (Cambridge research) during the reading process. Specific complementary tasks were administered to evaluate working memory, long-term memory, and executive functions. Variances analyses showed significantly lower performance by older adults regarding: 1) recall of the microstructure of the two texts with a high degree of detail, 2) macrostructure of the text with fewer details, and 3) performance on all tasks that evaluated cognitive components. Aging influenced treatment of levels of text representation depending on text characteristics. However, cluster analysis of the text comprehension and eye-tracker data revealed a group of older adults whose performance in reading comprehension was identical to the performance of younger adults, with the same reading profile. This result seems to show that use of compensation strategies by older adults at the onset of signs of cognitive deterioration is not necessary in reading. PMID- 17412666 TI - [Impairment of executive function in elderly patients with major unipolar depression: influence of psychomotor retardation]. AB - The results from several studies assessing the executive function in depressed patients compared to control subjects varied from significant impairment to normal performance. To assess the executive impairment in elderly patients with major unipolar depression and to evaluate the influence of psychomotor retardation and severity of depression in the executive deficits, the performance of 15 elderly patients with unipolar depression was compared to that of 15 elderly control subjects on executive tasks. The severity of depression was evaluated by the Montgomery and Asberg depressive scale and that of psychomotor retardation by the Widlocher's scale. In depressed patients, deficits were found on tasks assessing cognitive flexibility (Modified card sorting test (MCST) and Trail making test B), planification and elaboration of strategies (cognitive estimates), motor initiation (graphic sequences), categorisation and hypothesis making (MCST) and interference resistance (Stroop test). However, depressed patients performed normally on the Hayling test assessing the inhibition processes. Intensity of psychomotor retardation was not correlated to the performance of executive tasks. Conversely, severity of depression was related to the scores of MCST (number of errors and perseverations), Stroop and Hayling tests (time taken to complete the end of the sentence). Unipolar depressed patients showed deficits in most tasks assessing executive function. However, inhibition processes appeared to be intact in depressed patients although their implementation was difficult. The severity of depression but not that of psychomotor retardation was associated with executive deficits. PMID- 17412667 TI - Can predation by invasive mice drive seabird extinctions? AB - The house mouse, Mus musculus, is one of the most widespread and well-studied invasive mammals on islands. It was thought to pose little risk to seabirds, but video evidence from Gough Island, South Atlantic Ocean shows house mice killing chicks of two IUCN-listed seabird species. Mouse-induced mortality in 2004 was a significant cause of extremely poor breeding success for Tristan albatrosses, Diomedea dabbenena (0.27 fledglings/pair), and Atlantic petrels, Pterodroma incerta (0.33). Population models show that these levels of predation are sufficient to cause population decreases. Unlike many other islands, mice are the only introduced mammals on Gough Island. However, restoration programmes to eradicate rats and other introduced mammals from islands are increasing the number of islands where mice are the sole alien mammals. If these mouse populations are released from the ecological effects of predators and competitors, they too may become predatory on seabird chicks. PMID- 17412668 TI - Control of reproductive dominance by the thelytoky gene in honeybees. AB - Differentiation into castes and reproductive division of labour are a characteristics of eusocial insects. Caste determination occurs at an early stage of larval development in social bees and is achieved via differential nutrition irrespective of the genotype. Workers are usually subordinate to the queen and altruistically refrain from reproduction. Workers of the Cape honeybee (Apis mellifera capensis) do not necessarily refrain from reproduction. They have the unique ability to produce female offspring parthenogenetically (thelytoky) and can develop into 'pseudoqueens'. Although these are morphologically workers, they develop a queen-like phenotype with respect to physiology and behaviour. Thelytoky is determined by a single gene (th) and we show that this gene also influences other traits related to the queen phenotype, including egg production and queen pheromone synthesis. Using 566 microsatellite markers, we mapped this gene to chromosome 13 and identified a candidate locus thelytoky, similar to grainy head (a transcription factor), which has been shown to be highly expressed in queens of eusocial insects. We therefore suggest that this gene is not only important for determining the pseudoqueen phenotype in A. m. capensis workers, but is also of general importance in regulating the gene cascades controlling reproduction and sterility in female social bees. PMID- 17412669 TI - Southern Hemisphere humpback whales wintering off Central America: insights from water temperature into the longest mammalian migration. AB - We report on a wintering area off the Pacific coast of Central America for humpback whales (Megaptera novaeangliae) migrating from feeding areas off Antarctica. We document seven individuals, including a mother/calf pair, that made this migration (approx. 8300km), the longest movement undertaken by any mammal. Whales were observed as far north as 11 degrees N off Costa Rica, in an area also used by a boreal population during the opposite winter season, resulting in unique spatial overlap between Northern and Southern Hemisphere populations. The occurrence of such a northerly wintering area is coincident with the development of an equatorial tongue of cold water in the eastern South Pacific, a pattern that is repeated in the eastern South Atlantic. A survey of location and water temperature at the wintering areas worldwide indicates that they are found in warm waters (21.1-28.3 degrees C), irrespective of latitude. We contend that while availability of suitable reproductive habitat in the wintering areas is important at the fine scale, water temperature influences whale distribution at the basin scale. Calf development in warm water may lead to larger adult size and increased reproductive success, a strategy that supports the energy conservation hypothesis as a reason for migration. PMID- 17412670 TI - Spiders fluoresce variably across many taxa. AB - The evolution of fluorescence is largely unexplored, despite the newfound occurrence of this phenomenon in a variety of organisms. We document that spiders fluoresce under ultraviolet illumination, and find that the expression of this trait varies greatly among taxa in this species-rich group. All spiders we examined possess fluorophores in their haemolymph, but bright fluorescence appears to result when a spider sequesters fluorophores in its setae or cuticle. By sampling widely across spider taxa, we determine that fluorescent expression is labile and has evolved multiple times. Moreover, examination of the excitation and emission properties of extracted fluorophores reveals that spiders possess multiple fluorophores and that these differ among some families, indicating that novel fluorophores have evolved during spider diversification. Because many spiders fluoresce in wavelengths visible to their predators and prey (birds and insects), we propose that natural selection imposed by predator-prey interactions may drive the evolution of fluorescence in spiders. PMID- 17412671 TI - Early Cambrian record of failed durophagy and shell repair in an epibenthic mollusc. AB - Predation is arguably one of the main driving forces of early metazoan evolution, yet the fossil record of predation during the Ediacaran-Early Cambrian transition is relatively poor. Here, we present direct evidence of failed durophagous (shell breaking) predation and subsequent shell repair in the Early Cambrian (Botoman) epibenthic mollusc Marocella from the Mernmerna Formation and Oraparinna Shale in the Flinders Ranges, South Australia. This record pushes back the first appearance of durophagy on molluscs by approximately 40Myr. PMID- 17412672 TI - Intense ultrasonic clicks from echolocating toothed whales do not elicit anti predator responses or debilitate the squid Loligo pealeii. AB - Toothed whales use intense ultrasonic clicks to echolocate prey and it has been hypothesized that they also acoustically debilitate their prey with these intense sound pulses to facilitate capture. Cephalopods are an important food source for toothed whales, and there has probably been an evolutionary selection pressure on cephalopods to develop a mechanism for detecting and evading sound-emitting toothed whale predators. Ultrasonic detection has evolved in some insects to avoid echolocating bats, and it can be hypothesized that cephalopods might have evolved similar ultrasound detection as an anti-predation measure. We test this hypothesis in the squid Loligo pealeii in a playback experiment using intense echolocation clicks from two squid-eating toothed whale species. Twelve squid were exposed to clicks at two repetition rates (16 and 125 clicks per second) with received sound pressure levels of 199-226 dB re1 microPa (pp) mimicking the sound exposure from an echolocating toothed whale as it approaches and captures prey. We demonstrate that intense ultrasonic clicks do not elicit any detectable anti-predator behaviour in L. pealeii and that clicks with received levels up to 226 dB re1 microPa (pp) do not acoustically debilitate this cephalopod species. PMID- 17412673 TI - Death of an order: a comprehensive molecular phylogenetic study confirms that termites are eusocial cockroaches. AB - Termites are instantly recognizable mound-builders and house-eaters: their complex social lifestyles have made them incredibly successful throughout the tropics. Although known as 'white ants', they are not ants and their relationships with other insects remain unclear. Our molecular phylogenetic analyses, the most comprehensive yet attempted, show that termites are social cockroaches, no longer meriting being classified as a separate order (Isoptera) from the cockroaches (Blattodea). Instead, we propose that they should be treated as a family (Termitidae) of cockroaches. It is surprising to find that a group of wood-feeding cockroaches has evolved full sociality, as other ecologically dominant fully social insects (e.g. ants, social bees and social wasps) have evolved from solitary predatory wasps. PMID- 17412674 TI - Cytokinin-mediated leaf manipulation by a leafminer caterpillar. AB - A large number of hypotheses have been proposed to explain the adaptive significance and evolution of the endophagous-feeding mode, nutritional benefits being considered to be one of the main advantages. Leaf-mining insects should feed on most nutritional tissues and avoid tissues with high structural and/or biochemical plant defences. This selective feeding behaviour could furthermore be reinforced by manipulating the plant physiology, as suggested by the autumnal formation of 'green islands' around mining caterpillars in yellow leaves. The question we address here is how such metabolic manipulation occurs and what the nutritional consequences for the insect are. We report a large accumulation of cytokinins in the mined tissues which is responsible for the preservation of functional nutrient-rich green tissues at a time when leaves are otherwise turning yellow. The analogy with other plant manipulating organisms, in particular galling insects, is striking. PMID- 17412676 TI - Viscoelastic nanoscale properties of cuticle contribute to the high-pass properties of spider vibration receptor (Cupiennius salei Keys). AB - Atomic force microscopy (AFM) and surface force spectroscopy were applied in live spiders to their joint pad material located distal of the metatarsal lyriform organs, which are highly sensitive vibration sensors. The surface topography of the material is sufficiently smooth to probe the local nanomechanical properties with nanometre elastic deflections. Nanoscale loads were applied in the proximad direction on the distal joint region simulating the natural stimulus situation. The force curves obtained indicate the presence of a soft, liquid-like epicuticular layer (20-40 nm thick) above the pad material, which has much higher stiffness. The Young modulus of the pad material is close to 15 MPa at low frequencies, but increases rapidly with increasing frequencies approximately above 30 Hz to approximately 70 MPa at 112 Hz. The adhesive forces drop sharply by about 40% in the same frequency range. The strong frequency dependence of the elastic modulus indicates the viscoelastic nature of the pad material, its glass transition temperature being close to room temperature (25 +/- 2 degrees C) and, therefore, to its maximized energy absorption from low-frequency mechanical stimuli. These viscoelastic properties of the cuticular pad are suggested to be at least partly responsible for the high-pass characteristics of the vibration sensor's physiological properties demonstrated earlier. PMID- 17412677 TI - Transmissibility of swine flu at Fort Dix, 1976. AB - The 1976 outbreak of A/New Jersey/76 influenza in Fort Dix is a rare example of an influenza virus with documented human to human transmission that failed to spread widely. Despite extensive epidemiological investigation, no attempt has been made to quantify the transmissibility of this virus. The World Health Organization and the United States Government view containment of emerging influenza strains as central to combating pandemic influenza. Computational models predict that it may be possible to contain an emergent pandemic influenza if virus transmissibility is low. The A/New Jersey/76 outbreak at the United States Army Training Center at Fort Dix, New Jersey in January 1976 caused 13 hospitalizations, 1 death and an estimated 230 cases. To characterize viral transmission in this epidemic, we estimated the basic reproductive number and serial interval using deterministic epidemic models and stochastic simulations. We estimated the basic reproductive number for this outbreak to be 1.2 (supported interval 1.1-1.4), the serial interval to be 1.9 days (supported interval 1.6-3.8 days), and that the virus had at least six serial human to human transmissions. This places the transmissibility of A/New Jersey/76 virus at the lower end of circulating flu strains, well below the threshold for control. PMID- 17412675 TI - Natural origin biodegradable systems in tissue engineering and regenerative medicine: present status and some moving trends. AB - The fields of tissue engineering and regenerative medicine aim at promoting the regeneration of tissues or replacing failing or malfunctioning organs, by means of combining a scaffold/support material, adequate cells and bioactive molecules. Different materials have been proposed to be used as both three-dimensional porous scaffolds and hydrogel matrices for distinct tissue engineering strategies. Among them, polymers of natural origin are one of the most attractive options, mainly due to their similarities with the extracellular matrix (ECM), chemical versatility as well as typically good biological performance. In this review, the most studied and promising and recently proposed naturally derived polymers that have been suggested for tissue engineering applications are described. Different classes of such type of polymers and their blends with synthetic polymers are analysed, with special focus on polysaccharides and proteins, the systems that are more inspired by the ECM. The adaptation of conventional methods or non-conventional processing techniques for processing scaffolds from natural origin based polymers is reviewed. The use of particles, membranes and injectable systems from such kind of materials is also overviewed, especially what concerns the present status of the research that should lead towards their final application. Finally, the biological performance of tissue engineering constructs based on natural-based polymers is discussed, using several examples for different clinically relevant applications. PMID- 17412678 TI - Shifting and stopping: fronto-striatal substrates, neurochemical modulation and clinical implications. AB - The neuropsychological basis of attentional set-shifting, task-set switching and stop-signal inhibition is reviewed through comparative studies of humans and experimental animals. Using human functional neuroimaging, plus neuropsychological investigation of patients with frontal damage quantified by structural magnetic resonance imaging, and through parallels with effects of specific lesions of the prefrontal cortex (PFC) and striatum in rats and marmosets, it is possible to define both distinct and overlapping loci for tasks such as extra-dimensional shifting and reversal learning, stop-signal reaction time and task-set switching. Notably, most of the paradigms implicate a locus in the right PFC, specifically the right inferior frontal gyrus, possibly associated with processes of response inhibition. The neurochemical modulation of fronto striatal circuitry in parallel with effects on task performance has been investigated using specific neuropharmacological agents in animals and by human psychopharmacological investigations, sometimes in conjunction with functional imaging. Evidence is provided for double dissociations of effects of manipulations of prefrontal cortical catecholamine and indoleamine (5-HT) systems that have considerable implications in the treatment of disorders such as Parkinson's disease, attention deficit/hyperactivity disorder and depression, as well as in theoretical notions of how 'fronto-executive' functions are subject to state-dependent influences, probably related to stress, arousal and motivation. PMID- 17412679 TI - Is there a dysexecutive syndrome? AB - The role of the frontal lobes has often been described as a 'paradox' or a 'riddle'. Ascribed to this region has been the loftiest of functions (e.g. executive; seat of wisdom); others contested that the frontal lobes played no special role. There has also been controversy about the unity or diversity of functions related to the frontal lobes. Based on the analysis of the effects of lesions of the frontal lobes, we propose that there are discrete categories of functions within the frontal lobes, of which 'executive' functioning is one. Within the executive category, the data do not support the concept of an undifferentiated central executive/supervisory system. The results are better explained as impairments in a collection of anatomically and functionally independent but interrelated attentional control processes. Evidence for three separate frontal attentional processes is presented. For each process, we present an operational description, the data supporting the distinctiveness of each process and the evidence for impairments of each process after lesions in specific frontal regions. These processes and their coarse frontal localizations are energization-superior medial, task setting-left lateral and monitoring-right lateral. The strength of the findings lies in replication: across different tasks; across different cognitive modalities (e.g. reaction time paradigms, memory); and across different patient groups. This convergence minimizes the possibility that any of the findings are limited to a specific task or to a specific set of patients. Although distinct, these processes are flexibly assembled in response to context, complexity and intention over real time into different networks within the frontal regions and between frontal and posterior regions. PMID- 17412680 TI - Beyond the sentence given. AB - A central and influential idea among researchers of language is that our language faculty is organized according to Fregean compositionality, which states that the meaning of an utterance is a function of the meaning of its parts and of the syntactic rules by which these parts are combined. Since the domain of syntactic rules is the sentence, the implication of this idea is that language interpretation takes place in a two-step fashion. First, the meaning of a sentence is computed. In a second step, the sentence meaning is integrated with information from prior discourse, world knowledge, information about the speaker and semantic information from extra-linguistic domains such as co-speech gestures or the visual world. Here, we present results from recordings of event-related brain potentials that are inconsistent with this classical two-step model of language interpretation. Our data support a one-step model in which knowledge about the context and the world, concomitant information from other modalities, and the speaker are brought to bear immediately, by the same fast-acting brain system that combines the meanings of individual words into a message-level representation. Underlying the one-step model is the immediacy assumption, according to which all available information will immediately be used to co determine the interpretation of the speaker's message. Functional magnetic resonance imaging data that we collected indicate that Broca's area plays an important role in semantic unification. Language comprehension involves the rapid incorporation of information in a 'single unification space', coming from a broader range of cognitive domains than presupposed in the standard two-step model of interpretation. PMID- 17412681 TI - Quantifying the risk from ovine BSE and the impact of control strategies. AB - Although no naturally infected sheep with bovine spongiform encephalopathy (BSE) has ever been discovered, it remains possible that BSE once infected the UK sheep population, has been transmitted between sheep, and is still present today. We constructed a mathematical model to assess the current maximum theoretical exposure to consumers from BSE-infected ovine material and to estimate the risk reduction that could be achieved by abattoir-based control options if BSE infected sheep were ever found in the national flock. We predict that, if present, the exposure to consumers from a single BSE-infected sheep would be high: one sheep, close to the end of its incubation period, is likely to contribute 10-1000 times more infectious material than a fully infectious cow. Furthermore, 30% of this exposure comes from infectivity residing in lymphatic and peripheral tissue that cannot be completely removed from a carcass. We are 95% confident that throughout Great Britain, no more than four sheep flocks currently harbour an ongoing BSE epidemic. However, since the exposure from a single infected sheep is high, the annual human exposure from four 'typical' BSE infected flocks could be considerable. Small reductions in exposure could be achieved by strategies based on tissue testing, a 12-month age restriction or expanded definitions of high-risk tissues. A six-month age restriction is likely to be more effective and genotype-based strategies the most effective. PMID- 17412682 TI - Stoats (Mustela erminea) provide evidence of natural overland colonization of Ireland. AB - The current Irish biota has controversial origins. Ireland was largely covered by ice at the Last Glacial Maximum (LGM) and may not have had land connections to continental Europe and Britain thereafter. Given the potential difficulty for terrestrial species to colonize Ireland except by human introduction, we investigated the stoat (Mustela erminea) as a possible cold-tolerant model species for natural colonization of Ireland at the LGM itself. The stoat currently lives in Ireland and Britain and across much of the Holarctic region including the high Arctic. We studied mitochondrial DNA variation (1771 bp) over the whole geographical range of the stoat (186 individuals and 142 localities), but with particular emphasis on the British Isles and continental Europe. Irish stoats showed considerably greater nucleotide and haplotype diversity than those in Britain. Bayesian dating is consistent with an LGM colonization of Ireland and suggests that Britain was colonized later. This later colonization probably reflects a replacement event, which can explain why Irish and British stoats belong to different mitochondrial lineages as well as different morphologically defined subspecies. The molecular data strongly indicate that stoats colonized Ireland naturally and that their genetic variability reflects accumulation of mutations during a population expansion on the island. PMID- 17412683 TI - Birth order and male androphilia in Samoan fa'afafine. AB - The manner in which male androphilia is publicly expressed varies cross culturally. As such, it is unclear whether distinct or common underlying causal processes characterize male androphilia in different cultures. Establishing the existence of cross-cultural universals in male androphilia is one means of ascertaining whether common biological bases underlie this phenomenon despite its culturally distinct forms. The evidence that the number of older biological brothers increases the odds of androphilia in later-born males has been well documented for Western samples (i.e. the fraternal birth order effect); but there is little evidence for this effect in non-Western samples. Here, we compare the birth order of androphilic males (i.e. fa'afafine) and gynephilic males from the politically autonomous Polynesian nation of Independent Samoa. Results indicate that relative to gynephilic males, fa'afafine tend to have more siblings and are generally later born when birth order is quantified using Slater, fraternal and sororal indices. More specifically, fa'afafine tend to have a greater number of older brothers, older sisters and younger brothers. We discuss the observed effects in relation to the differing reproductive patterns exhibited by the mothers of fa'afafine and gynephilic males, and to existing social and biological theories for sexual orientation. PMID- 17412684 TI - Benefits to shorebirds from invasion of a non-native shellfish. AB - Introductions of non-native species are seen as major threats to ecosystem function and biodiversity. However, invasions of aquatic habitats by non-native species are known to benefit generalist consumers that exhibit dietary switches and prey upon the exotic species in addition to or in preference to native ones. There is, however, little knowledge concerning the population-level implications of such dietary changes. Here, we show that the introduction of the Manila clam Tapes philippinarum into European coastal waters has presented the Eurasian oystercatcher Haematopus ostralegus ostralegus with a new food resource and resulted in a previously unknown predator-prey interaction between these species. We demonstrate, with an individuals-based simulation model, that the presence of this non-native shellfish, even at the current low density, has reduced the predicted over-winter mortality of oystercatchers at one recently invaded site. Further increases in clam population density are predicted to have even more pronounced effects on the density dependence of oystercatcher over-winter mortality. These results suggest that if the Manila clam were to spread around European coastal waters, a process which is likely to be facilitated by global warming, this could have considerable benefits for many shellfish-eating shorebird populations. PMID- 17412685 TI - Mitochondrial DNA analysis shows a Near Eastern Neolithic origin for domestic cattle and no indication of domestication of European aurochs. AB - The extinct aurochs (Bos primigenius primigenius) was a large type of cattle that ranged over almost the whole Eurasian continent. The aurochs is the wild progenitor of modern cattle, but it is unclear whether European aurochs contributed to this process. To provide new insights into the demographic history of aurochs and domestic cattle, we have generated high-confidence mitochondrial DNA sequences from 59 archaeological skeletal finds, which were attributed to wild European cattle populations based on their chronological date and/or morphology. All pre-Neolithic aurochs belonged to the previously designated P haplogroup, indicating that this represents the Late Glacial Central European signature. We also report one new and highly divergent haplotype in a Neolithic aurochs sample from Germany, which points to greater variability during the Pleistocene. Furthermore, the Neolithic and Bronze Age samples that were classified with confidence as European aurochs using morphological criteria all carry P haplotype mitochondrial DNA, suggesting continuity of Late Glacial and Early Holocene aurochs populations in Europe. Bayesian analysis indicates that recent population growth gives a significantly better fit to our data than a constant-sized population, an observation consistent with a postglacial expansion scenario, possibly from a single European refugial population. Previous work has shown that most ancient and modern European domestic cattle carry haplotypes previously designated T. This, in combination with our new finding of a T haplotype in a very Early Neolithic site in Syria, lends persuasive support to a scenario whereby gracile Near Eastern domestic populations, carrying predominantly T haplotypes, replaced P haplotype-carrying robust autochthonous aurochs populations in Europe, from the Early Neolithic onward. During the period of coexistence, it appears that domestic cattle were kept separate from wild aurochs and introgression was extremely rare. PMID- 17412686 TI - Reproductive failure in patients with various percentages of macronuclear spermatozoa: high level of aneuploid and polyploid spermatozoa. AB - The aim of this study was to describe the association between various percentages of macronuclear spermatozoa (MNSs), sperm chromosomal abnormalities, and reproductive failure in 4 patients. One patient had a familial history of perinatal deaths. Patients were selected according to the coexistence of normal sized spermatozoa and MNSs (19%, 22%, 29.5%, and 49.7%). Fluorescent in situ hybridization (FISH) on spermatozoa and semiautomated analysis of nuclear surface were assessed. All patients were characterized by an oligoasthenozoospermia. Three patients had a prevalence of irregular MNSs and prevalence of nondisjunction at the first meiotic division. One patient had a prevalence of regular MNSs and a prevalence of nondisjunction at the second meiotic division. FISH also showed a high rate of polyploidy and various rates of aneuploid sperm. The percentage of sperm with abnormal chromosome complements (25.6%, 43.6%, 51.4%, 71.7% with 3-color FISH) was higher than the percentage of MNSs. A population of apparently normal-sized spermatozoa that could be used for intracytoplasmic sperm injection (ICSI) was aneuploid. Sperm nuclear surface analysis revealed either a shift toward elevated values or distinguished 2 sperm subpopulations: normal and macronuclear. Patients underwent 7 ICSI cycles. The fertilization rate was low for 3 patients (50%, 40%, 50%) and normal for 1 patient (83.3%). Pregnancy rate per transfer was low (14.3%). The present study shows that the macronuclear phenotype can manifest a variety of clinical aspects. It is also shown that mild rates of MNSs impair fertility and constitute a risk of chromosomal abnormality for the embryos and a risk of perinatal death. We suggest conducting FISH on spermatozoa and genetic counseling for a couple when the percentage of MNSs reaches 20% in at least 1 spermiogram. PMID- 17412687 TI - XB130, a novel adaptor protein for signal transduction. AB - Adaptor proteins are important mediators in signal transduction. In the present study, we report the cloning and characterization of a novel adaptor protein, XB130. This gene is located on human chromosome 10q25.3 and encodes a protein of 818 amino acids. It contains several Src homology (SH)2- and SH3-binding motifs, two pleckstrin homology domains, a coiled-coil region, and a number of potential tyrosine or serine/threonine phosphorylation sites. Endogenous XB130 interacts with c-Src tyrosine kinase. Their co-expression in COS-7 cells resulted in activation of c-Src and elevated tyrosine phosphorylation of multiple proteins, including XB130 itself. XB130 expression in HEK293 cells enhanced serum response element- and AP-1-dependent transcriptional activation mediated by c-Src. XB130DeltaN, an N-terminal deletion mutant lacking a putative SH3-binding motif and several putative SH2-binding sites, reduced its ability to mediate Src signal transduction. Down-regulation of endogenous XB130 with siRNA reduced c-Src activity, IL-8 production, EGF-induced phosphorylation of Akt and GSK3beta, and altered cell cycles in human lung epithelial cells. These data suggest that XB130 as an adaptor may play an important role in the regulation of signal transduction and cellular functions. PMID- 17412688 TI - Directed evolution of ribosomal protein S1 for enhanced translational efficiency of high GC Rhodopseudomonas palustris DNA in Escherichia coli. AB - The expression of foreign DNA in Escherichia coli is important in biotechnological applications. However, the translation of genes from GC-rich organisms is inefficient in E. coli. To overcome this problem, we applied directed evolution to E. coli ribosomal protein S1. Two selected mutants enabled 12- and 8-fold higher expression levels from GC-rich DNA targets. General improvements in translation efficiency over a range of genes from Rhodopseudomonas palustris and E. coli was achieved using an S1 mutant selected against multiple genes from R. palustris. This method opens new opportunities for the expression of GC-rich genes in E. coli. PMID- 17412689 TI - Specific protein-membrane contacts are required for prepore and pore assembly by a cholesterol-dependent cytolysin. AB - Three short hydrophobic loops and a conserved undecapeptide at the tip of domain 4 (D4) of the cholesterol-dependent cytolysins (CDCs) mediate the binding of the CDC monomers to cholesterol-rich cell membranes. But intermedilysin (ILY), from Streptococcus intermedius, does not bind to cholesterol-rich membranes unless they contain the human protein CD59. This observation suggested that the D4 loops, which include loops L1-L3 and the undecapeptide, of ILY were no longer required for its cell binding. However, we show here that membrane insertion of the D4 loops is required for the cytolysis by ILY. Receptor binding triggers changes in the structure of ILY that are necessary for oligomerization, but membrane insertion of the D4 loops is critical for oligomer assembly and pore formation. Defects that prevent membrane insertion of the undecapeptide also block assembly of the prepore oligomer, while defects in the membrane insertion of the L1-L3 loops prevent the conversion of the prepore oligomer to the pore complex. These studies reveal that pore formation by ILY, and probably other CDCs, is affected by an intricate and coupled sequence of interactions between domain 4 and the membrane. PMID- 17412690 TI - Kinetic analysis of the entire RNA amplification process by Qbeta replicase. AB - The kinetics of the RNA replication reaction by Qbeta replicase were investigated. Qbeta replicase is an RNA-dependent RNA polymerase responsible for replicating the RNA genome of coliphage Qbeta and plays a key role in the life cycle of the Qbeta phage. Although the RNA replication reaction using this enzyme has long been studied, a kinetic model that can describe the entire RNA amplification process has yet to be determined. In this study, we propose a kinetic model that is able to account for the entire RNA amplification process. The key to our proposed kinetic model is the consideration of nonproductive binding (i.e. binding of an enzyme to the RNA where the enzyme cannot initiate the reaction). By considering nonproductive binding and the notable enzyme inactivation we observed, the previous observations that remained unresolved could also be explained. Moreover, based on the kinetic model and the experimental results, we determined rate and equilibrium constants using template RNAs of various lengths. The proposed model and the obtained constants provide important information both for understanding the basis of Qbeta phage amplification and the applications using Qbeta replicase. PMID- 17412691 TI - Protein kinase A-regulated nucleocytoplasmic shuttling of Id1 during angiogenesis. AB - Id1, an inhibitory partner of basic-helix-loop-helix transcriptional factors, has recently been recognized as a potent contributor to angiogenesis. However, the molecular mechanism underlying its role in angiogenesis remains essentially unknown. Herein we demonstrate the subcellular localization of Id1 to be altered depending on the cellular context of vascular endothelial cells. Id1 was localized in the nuclei of human umbilical vein endothelial cells (HUVECs) cultured on uncoated plates, whereas it was translocated to the cytoplasm in HUVECs on Matrigel along with the formation of capillary-like structures. Treatment with the nuclear export inhibitor leptomycin B and mutagenesis analysis using green fluorescent protein-fused Id1 revealed CRM1/exportin-dependent nuclear export of Id1 in HUVECs on Matrigel. This nuclear export of Id1 was inhibited by protein kinase A (PKA) activation by dibutyryl cyclic AMP and forskolin but was promoted by PKA inactivation by H-89 and MDL-12,330A. Mutagenesis analysis of Id1 showed the phosphorylation of Ser-5 to possibly mediate the effect of PKA. These results suggest the function of Id1 as a transcriptional factor to be controlled by nucleocytoplasmic shuttling during angiogenesis and that PKA might be involved in this process. This may serve as a novel mechanism regulating angiogenesis and as a possible target for therapeutic vascular regeneration. PMID- 17412692 TI - Nucleoside phosphorylation by phosphate minerals. AB - In the presence of formamide, crystal phosphate minerals may act as phosphate donors to nucleosides, yielding both 5'- and, to a lesser extent, 3' phosphorylated forms. With the mineral Libethenite the formation of 5'-AMP can be as high as 6% of the adenosine input and last for at least 10(3) h. At high concentrations, soluble non-mineral phosphate donors (KH(2)PO(4) or 5'-CMP) afford 2'- and 2':3'-cyclic AMP in addition to 5'-and 3'-AMP. The phosphate minerals analyzed were Herderite Ca[BePO(4)F], Hureaulite Mn(2+)(5)(PO(3)(OH)(2)(PO(4))(2)(H(2)O)(4), Libethenite Cu(2+)(2)(PO(4))(OH), Pyromorphite Pb(5)(PO(4))(3)Cl, Turquoise Cu(2+)Al(6)(PO(4))(4)(OH)(8)(H(2)O)(4), Fluorapatite Ca(5)(PO(4))(3)F, Hydroxylapatite Ca(5)(PO(4))(3)OH, Vivianite Fe(2+)(3)(PO(4))(2)(H(2)O)(8), Cornetite Cu(2+)(3)(PO(4))(OH)(3), Pseudomalachite Cu(2+)(5)(PO(4))(2)(OH)(4), Reichenbachite Cu(2+)(5)(PO(4))(2)(OH)(4), and Ludjibaite Cu(2+)(5)(PO(4))(2)(OH)(4)). Based on their behavior in the formamide driven nucleoside phosphorylation reaction, these minerals can be characterized as: 1) inactive, 2) low level phosphorylating agents, or 3) active phosphorylating agents. Instances were detected (Libethenite and Hydroxylapatite) in which phosphorylation occurs on the mineral surface, followed by release of the phosphorylated compounds. Libethenite and Cornetite markedly protect the beta glycosidic bond. Thus, activated nucleic monomers can form in a liquid non aqueous environment in conditions compatible with the thermodynamics of polymerization, providing a solution to the standard-state Gibbs free energy change (DeltaG degrees ') problem, the major obstacle for polymerizations in the liquid phase in plausible prebiotic scenarios. PMID- 17412693 TI - Munc18c interaction with syntaxin 4 monomers and SNARE complex intermediates in GLUT4 vesicle trafficking. AB - In the process of insulin-stimulated GLUT4 vesicle exocytosis, Munc18c has been proposed to control SNARE complex formation by inactivating syntaxin 4 in a self associated conformation. Using in vivo fluorescence resonance energy transfer in 3T3L1 adipocytes, co-immunoprecipitation, and in vitro binding assays, we provide data to indicate that Munc18c also associates with nearly equal affinity to a mutant of syntaxin 4 in a constitutively open (unfolded) state (L173A/E174A; LE). To bind to the open conformation of syntaxin 4, we found that Munc18c requires an interaction with the N terminus of syntaxin 4, which resembles Sly1 interaction with the N terminus of ER/Golgi syntaxins. However, both N and C termini of syntaxin 4 are required for Munc18c binding, since a mutation in the syntaxin 4 SNARE domain (I241A) reduces the interaction, irrespective of syntaxin 4 conformation. Using an optical reporter for syntaxin 4-SNARE pairings in vivo, we demonstrate that Munc18c blocks recruitment of SNAP23 to wild type syntaxin 4 yet associates with syntaxin 4LE-SNAP23 Q-SNARE complexes. Fluorescent imaging of GLUT4 vesicles in 3T3L1 adipocytes revealed that syntaxin 4LE expressed with Munc18c bypasses the requirement of insulin for GLUT4 vesicle plasma membrane docking. This effect was attenuated by reducing the Munc18c-syntaxin 4LE interaction with the I241A mutation, indicating that Munc18c facilitates vesicle docking. Therefore, in contradiction to previous models, our data indicates that the conformational "opening" of syntaxin 4 rather than the dissociation of Munc18c is the critical event required for GLUT4 vesicle docking. PMID- 17412694 TI - Phosphorylation of p68 RNA helicase plays a role in platelet-derived growth factor-induced cell proliferation by up-regulating cyclin D1 and c-Myc expression. AB - p68 RNA helicase is a protypical member of DEAD box family RNA helicase. The protein plays an important role in the cell developmental program and organ maturation. We demonstrated previously that, in response to growth factor platelet-derived growth factor (PDGF)-BB stimulation, p68 is phosphorylated at Tyr(593), and the phosphorylation of p68 promotes epithelial-mesenchymal transition via promoting beta-catenin nuclear translocation (Yang, L., Lin, C., and Liu, Z. R. (2006) Cell 127, 139-155). We show here that the tyrosine phosphorylation of p68 also mediates the effects of PDGF in stimulating cell proliferation. The phosphorylated p68 (referred to as phospho-p68) promotes cell proliferation by activating the transcription of cyclin D1 and c-Myc genes. We show that the ATPase/helicase activities of p68 are required for the activation of cyclin D1 transcription. The phospho-p68 participates in the complex assembled at the cyclin D1 and c-Myc promoters, which strongly suggests a direct role in transcriptional regulation. Furthermore, our data demonstrated that the phosphorylation of p68 at Tyr(593) plays a role in mediating the autocrine loop effects of PDGF, suggesting an important role for p68 phosphorylation in cell proliferation. PMID- 17412695 TI - EHD1 and Eps15 interact with phosphatidylinositols via their Eps15 homology domains. AB - The C-terminal Eps15 homology domain-containing protein, EHD1, regulates the recycling of receptors from the endocytic recycling compartment to the plasma membrane. In cells, EHD1 localizes to tubular and spherical recycling endosomes. To date, the mode by which EHD1 associates with endosomal membranes remains unknown, and it has not been determined whether this interaction is direct or via interacting proteins. Here, we provide evidence demonstrating that EHD1 has the ability to bind directly and preferentially to an array of phospholipids, preferring phosphatidylinositols with a phosphate at position 3. Previous studies have demonstrated that EH domains coordinate calcium binding and interact with proteins containing the tripeptide asparagine-proline-phenylalanine (NPF). Using two-dimensional nuclear magnetic resonance analysis, we now describe a new function for the Eps15 homology (EH) domain of EHD1 and show that it is capable of directly binding phosphatidylinositol moieties. Moreover, we have expanded our studies to include the C-terminal EH domain of EHD4 and the second of the three N terminal EH domains of Eps15 and demonstrated that phosphatidylinositol binding may be a more general property shared by certain other EH domains. Further studies identified a positively charged lysine residue (Lys-483) localized within the third helix of the EH domain, on the opposite face of the NPF-binding pocket, as being critical for the interaction with the phosphatidylinositols. PMID- 17412696 TI - Unique dimeric structure of BNip3 transmembrane domain suggests membrane permeabilization as a cell death trigger. AB - BNip3 is a prominent representative of apoptotic Bcl-2 proteins with rather unique properties initiating an atypical programmed cell death pathway resembling both necrosis and apoptosis. Many Bcl-2 family proteins modulate the permeability state of the outer mitochondrial membrane by forming homo- and hetero-oligomers. The structure and dynamics of the homodimeric transmembrane domain of BNip3 were investigated with the aid of solution NMR in lipid bicelles and molecular dynamics energy relaxation in an explicit lipid bilayer. The right-handed parallel helix-helix structure of the domain with a hydrogen bond-rich His-Ser node in the middle of the membrane, accessibility of the node for water, and continuous hydrophilic track across the membrane suggest that the domain can provide an ion-conducting pathway through the membrane. Incorporation of the BNip3 transmembrane domain into an artificial lipid bilayer resulted in pH dependent conductivity increase. A possible biological implication of the findings in relation to triggering necrosis-like cell death by BNip3 is discussed. PMID- 17412697 TI - Mutational and structural studies aimed at characterizing the monomer of HIV-1 protease and its precursor. AB - An experimental protocol for folding the mature human immunodeficiency virus-1 (HIV-1) protease is presented that facilitates NMR studies at a low protein concentration of approximately 20 micoM. Under these conditions, NMR spectra show that the mature protease lacking its terminal beta-sheet residues 1-4 and 96-99 (PR(5-95)) exhibits a stable monomer fold spanning the region 10-90 that is similar to that of the single subunit of the wild-type dimer and the dimer bearing a D25N mutation (PR(D25N)). Urea-induced unfolding monitored both by changes in (1)H-(15)N heteronuclear single quantum correlation spectra and by protein fluorescence indicates that although PR(5-95) monomer displays a transition profile similar to that of the PR(D25N) dimer (50% unfolded (U(50)) = approximately 1.9 M), extending the protease with 4 residues (SFNF) of its N terminally flanking sequence in the Gag-Pol precursor ((SFNF)PR(D25N)) decreases the stability of the fold (U(50) = approximately 1.5 M). Assigned backbone chemical shifts were used to elucidate differences in the stability of the PR(T26A) (U(50) = 2.5 M) and (SFNF)PR(D25N) monomers and compared with PR(D25N/T26A) monomer. Discernible differences in the backbone chemical shifts were observed for N-terminal protease residues 3-6 of (SFNF)PR(D25N) that may relate to the increase in the equilibrium dissociation constant (K(d)) and the very low catalytic activity of the protease prior to its autoprocessing at its N terminus from the Gag-Pol precursor. PMID- 17412699 TI - Renal AT 1 receptors and hypertension. PMID- 17412698 TI - SOX6 suppresses cyclin D1 promoter activity by interacting with beta-catenin and histone deacetylase 1, and its down-regulation induces pancreatic beta-cell proliferation. AB - Sex-determining region Y-box (SOX) 6 negatively regulates glucose-stimulated insulin secretion from beta-cells and is a down-regulated transcription factor in the pancreatic islet cells of hyperinsulinemic obese mice. To determine the contribution of SOX6 to insulin resistance, we analyzed the effects of SOX6 on cell proliferation. Small interfering RNA-mediated attenuation of SOX6 expression stimulated the proliferation of insulinoma INS-1E and NIH-3T3 cells, whereas retroviral overexpression resulted in inhibition of cell growth. Quantitative real time-PCR analysis revealed that the levels of cyclin D1 transcripts were markedly decreased by SOX6 overexpression. Luciferase-reporter assay with beta catenin showed that SOX6 suppresses cyclin D1 promoter activities. In vitro binding experiments showed that the LZ/Q domain of SOX6 physically interacts with armadillo repeats 1-4 of beta-catenin. Furthermore, chromatin immunoprecipitation assay revealed that increased SOX6 expression significantly reduced the levels of acetylated histones H3 and H4 at the cyclin D1 promoter. By using a histone deacetylase (HDAC) inhibitor and co-immunoprecipitation analysis, we showed that SOX6 suppressed cyclin D1 activities by interacting withbeta-catenin and HDAC1. The data presented suggest that SOX6 may be an important factor in obesity related insulin resistance. PMID- 17412700 TI - Prime time for polycystic kidney disease: does one shot of roscovitine bring the cure? PMID- 17412701 TI - Regional effects of satellite haemodialysis units on renal replacement therapy in non-urban Ontario, Canada. AB - BACKGROUND: To provide better dialysis care to rural communities, the Ministry of Health chose to build satellite haemodialysis (HD) units, which are affiliated with, but are distant to, a main renal centre. We considered whether constructing such units in rural regions of Ontario, Canada, alleviated under-service of rates of renal replacement therapy (RRT) locally, decreased patient travel distance and decreased local peritoneal dialysis (PD) utilization. METHODS: We compared two groups of rural regions at two time points (years 1995 and 2002) in a before and after cross-sectional study. These regions were either already serviced by a satellite unit in 1995 (control group, 10 communities), or had new satellite units built between the years 1995 and 2002 (exposure group, 24 communities). RESULTS: The exposure group had a slightly greater increase in prevalent rate of RRT over time, but this did not reach statistical significance (control group increased 401 per million, exposure group 436 per million, P = 0.8). The mean weekly travel distance was reduced by 210.6 km after the construction of new satellite units (P < 0.001). There was no significant difference between the groups in reduction of PD proportion (P = 0.4). There was a significant increase in the number of elderly receiving RRT once local access was provided. CONCLUSIONS: In conclusion, constructing satellite units increased access to renal care for elderly patients and reduced travel time for HD patients living in rural communities. PMID- 17412702 TI - Dialysis or not? A comparative survival study of patients over 75 years with chronic kidney disease stage 5. AB - BACKGROUND: The number of elderly patients with chronic kidney disease (CKD) stage 5 is steadily increasing. Evidence is needed to inform decision-making for or against dialysis, especially in those patients with multiple comorbid conditions for whom dialysis may not increase survival. We therefore compared survival of elderly patients with CKD stage 5, managed either with dialysis or conservatively (without dialysis), after the management decision had been made, and explored which of several key variables were independently associated with survival. METHODS: A retrospective analysis of the survival of all over 75 years with CKD stage 5 attending dedicated multidisciplinary pre-dialysis care clinics (n=129) was performed. Demographic and comorbidity data were collected on all patients. Survival was defined as the time from estimated GFR<15 ml/min to either death or study endpoint. RESULTS: One- and two-year survival rates were 84% and 76% in the dialysis group (n=52) and 68% and 47% in the conservative group (n=77), respectively, with significantly different cumulative survival (log rank 13.6, P<0.001). However, this survival advantage was lost in those patients with high comorbidity scores, especially when the comorbidity included ischaemic heart disease. CONCLUSIONS: In CKD stage 5 patients over 75 years, who receive specialist nephrological care early, and who follow a planned management pathway, the survival advantage of dialysis is substantially reduced by comorbidity and ischaemic heart disease in particular. Comorbidity should be a major consideration when advising elderly patients for or against dialysis. PMID- 17412703 TI - Coexistence of tubulins and ftsZ in different Prosthecobacter species. AB - Prosthecobacter, one of the few cultivable representatives of the bacterial phylum Verrucomicrobia, is of increasing interest to the scientific community due to the presence of tubulin genes in its genome and the apparent absence of the bacterial homologue FtsZ that is normally involved in prokaryotic cell division. These findings suggested the possibility of a vicarious takeover of the FtsZ function through these novel tubulins and opened new scenarios on the possible evolution of bacterial cytoskeleton and cell division. In the present manuscript, we report the characterization of ftsZ and ftsA homologues in different Prosthecobacter species that also possess tubulin genes. Based on these findings, we propose an FtsZ-based cell division mechanism in Verrucomicrobia. The analysis of available genome data of Verrucomicrobia suggests that tubulins are not a feature common to all members of this phylum. Therefore, it can be assumed that Prosthecobacter acquired tubulins through horizontal gene transfer. The functional role of tubulins in Prosthecobacter remains enigmatic. PMID- 17412704 TI - Base-flipping dynamics in a DNA hairpin processing reaction. AB - Many enzymes that repair or modify bases in double-stranded DNA gain access to their substrates by base flipping. Although crystal structures provide stunning snap shots, biochemical approaches addressing the dynamics have proven difficult, particularly in complicated multi-step reactions. Here, we use protein-DNA crosslinking and potassium permanganate reactivity to explore the base-flipping step in Tn5 transposition. We present a model to suggest that base flipping is driven by a combination of factors including DNA bending and the intrusion of a probe residue. The forces are postulated to act early in the reaction to create a state of tension, relieved by base flipping after cleavage of the first strand of DNA at the transposon end. Elimination of the probe residue retards the kinetics of nicking and reduces base flipping by 50%. Unexpectedly, the probe residue is even more important during the hairpin resolution step. Overall, base flipping is pivotal to the hairpin processing reaction because it performs two opposite but closely related functions. On one hand it disrupts the double helix, providing the necessary strand separation and steric freedom. While on the other, transposase appears to position the second DNA strand in the active site for cleavage using the flipped base as a handle. PMID- 17412705 TI - Determination of the DNA-binding kinetics of three related but heteroimmune bacteriophage repressors using EMSA and SPR analysis. AB - Bacteriophages P2, P2 Hy dis and WPhi are very similar but heteroimmune Escherichia coli phages. The structural genes show over 96% identity, but the repressors show between 43 and 63% identities. Furthermore, the operators, which contain two directly repeated sequences, vary in sequence, length, location relative to the promoter and spacing between the direct repeats. We have compared the in vivo effects of the wild type and mutated operators on gene expression with the complexes formed between the repressors and their wild type or mutated operators using electrophoretic mobility shift assay (EMSA), and real-time kinetics of the protein-DNA interactions using surface plasmon resonance (SPR) analysis. Using EMSA, the repressors formed different protein-DNA complexes, and only WPhi was significantly affected by point mutations. However, SPR analysis showed a reduced association rate constant and an increased dissociation rate constant for P2 and WPhi operator mutants. The association rate constants of P2 Hy dis was too fast to be determined. The P2 Hy dis dissociation response curves were shown to be triphasic, while both P2 and WPhi C were biphasic. Thus, the kinetics of complex formation and the nature of the complexes formed differ extensively between these very closely related phages. PMID- 17412706 TI - Concentration-dependent organization of DNA by the dinoflagellate histone-like protein HCc3. AB - The liquid crystalline chromosomes of dinoflagellates are the alternative to the nucleosome-based organization of chromosomes in the eukaryotes. These nucleosome less chromosomes have to devise novel ways to maintain active parts of the genome. The dinoflagellate histone-like protein HCc3 has significant sequence identity with the bacterial DNA-binding protein HU. HCc3 also has a secondary structure resembling HU in silico. We have examined HCc3 in its recombinant form. Experiments on DNA-cellulose revealed its DNA-binding activity is on the C terminal domain. The N-terminal domain is responsible for intermolecular oligomerization as demonstrated by cross-linking studies. However, HCc3 could not complement Escherichia coli HU-deficient mutants, suggesting functional differences. In ligation assays, HCc3-induced DNA concatenation but not ring closure as the DNA-bending HU does. The basic HCc3 was an efficient DNA condensing agent, but it did not behave like an ordinary polycationic compound. HCc3 also induced specific structures with DNA in a concentration-dependent manner, as demonstrated by atomic force microscopy (AFM). At moderate concentration of HCc3, DNA bridging and bundling were observed; at high concentrations, the complexes were even more condensed. These results are consistent with a biophysical role for HCc3 in maintaining extended DNA loops at the periphery of liquid crystalline chromosomes. PMID- 17412707 TI - C1D and hMtr4p associate with the human exosome subunit PM/Scl-100 and are involved in pre-rRNA processing. AB - The exosome is a complex of 3'-5' exoribonucleases and RNA-binding proteins, which is involved in processing or degradation of different classes of RNA. Previously, the characterization of purified exosome complexes from yeast and human cells suggested that C1D and KIAA0052/hMtr4p are associated with the exosome and thus might regulate its functional activities. Subcellular localization experiments demonstrated that C1D and KIAA0052/hMtr4p co-localize with exosome subunit PM/Scl-100 in the nucleoli of HEp-2 cells. Additionally, the nucleolar accumulation of C1D appeared to be dependent on PM/Scl-100. Protein protein interaction studies showed that C1D binds to PM/Scl-100, whereas KIAA0052/hMtr4p was found to interact with MPP6, a previously identified exosome associated protein. Moreover, we demonstrate that C1D, MPP6 and PM/Scl-100 form a stable trimeric complex in vitro. Knock-down of C1D, MPP6 and KIAA0052/hMtr4p by RNAi resulted in the accumulation of 3'-extended 5.8S rRNA precursors, showing that these proteins are required for rRNA processing. Interestingly, C1D appeared to contain RNA-binding activity with a potential preference for structured RNAs. Taken together, our results are consistent with a role for the exosome-associated proteins C1D, MPP6 and KIAA052/hMtr4p in the recruitment of the exosome to pre rRNA to mediate the 3' end processing of the 5.8S rRNA. PMID- 17412708 TI - NMR evaluation of ammonium ion movement within a unimolecular G-quadruplex in solution. AB - d[G4(T4G4)3] has been folded into a unimolecular G-quadruplex in the presence of 15NH4+ ions. NMR spectroscopy confirmed that its topology is the same as the solution state structure determined earlier by Wang and Patel (J. Mol. Biol., 1995; 251: 76-94) in the presence of Na+ ions. The d[G4(T4G4)3] G-quadruplex exhibits four G-quartets with three 15NH4+-ion-binding sites (O1, I and O2). Quantitative analysis utilizing 15NH4+ ions as a NMR probe clearly demonstrates that there is no unidirectional 15NH4+ ion movement through the central cavity of the G-quadruplex. 15NH4+ ions move back and forth between the binding sites within the G-quadruplex and exchange with ions in bulk solution. 15NH4+ ion movement is controlled by the thermodynamic preferences of individual binding sites, steric restraints of the G-quartets for 15NH4+ ion passage and diagonal versus edge-type arrangement of the T4 loops. The movement of 15NH4+ ions from the interior of the G-quadruplex to bulk solution is faster than exchange within the G-quadruplex. The structural details of the G-quadruplex define stiffness of individual G-quartets that intimately affects 15NH4+ ion movement. The stiffness of G-quartets and steric hindrance imposed by thymine residues in the loops contribute to the 5-fold difference in the exchange rate constants through the outer G-quartets. PMID- 17412709 TI - Evidence-based therapeutics--introducing the Cochrane corner. PMID- 17412710 TI - Is COMT a susceptibility gene for schizophrenia? AB - Catechol-O-methyl transferase (COMT) is a catabolic enzyme involved in the degradation of a number of bioactive molecules; of principal interest to psychiatry, these include dopamine. The enzyme is encoded by the COMT gene. COMT is located (along with 47 other genes) in a fragment of chromosome 22q11 which when deleted results in a complex syndrome, the psychiatric manifestations of which include schizophrenia and other psychoses. These 2 observations have placed COMT near the top of a rather long list of plausible candidate genes for schizophrenia. The ability to test the hypothesis that COMT might be a susceptibility gene for schizophrenia has been simplified in principle by the existence of a valine-to-methionine (Val/Met) polymorphism which results respectively in high and low activity forms of the enzyme. Given the unequivocal effect of this polymorphism on the function of COMT, and the evidence for a critical role for dopamine in the pathophysiology and treatment of psychosis, there are strong prior expectations that Val/Met influences susceptibility to schizophrenia as well as other psychiatric phenotypes. Indeed the Val/Met polymorphism has become the most widely studied polymorphism in psychiatry. In this review, we consider the evidence for and against the involvement of COMT in schizophrenia. The current data allow us to virtually exclude a simple relationship between schizophrenia and the Val/Met variant previously thought to dominate COMT function. However, recent data suggest a more complex pattern of genetic regulation of COMT function beyond that attributable to the Val/Met locus. Moreover, it is also clear that there is a complex nonlinear relationship between dopamine availability and brain function. These 2 factors, allied to phenotypic complexity within schizophrenia, make it difficult to draw strong conclusions regarding COMT in schizophrenia. Nevertheless, emerging research that takes greater account of all these levels of complexity is beginning to provide tantalizing, but far from definitive, support for the view that COMT influences susceptibility to at least some forms of psychosis. PMID- 17412711 TI - Cognitive functioning in the schizophrenia prodrome. AB - In the last decade, there has been an increasing interest in cognitive alterations during the early course of schizophrenia. From a clinical perspective, a better understanding of cognitive functioning in putative at-risk states for schizophrenia is essential for developing optimal early intervention models. Two approaches have more recently been combined to assess the entire course of the initial schizophrenia prodrome: the predictive "basic symptom at risk" (BS) and the ultra high-risk (UHR) criteria. Basic symptoms are considered to be present during the entire disease progression, including the initial prodrome, while the onset of symptoms captured by the UHR criteria expresses further disease progression toward frank psychosis. The present study investigated the cognitive functioning in 93 subjects who met either BS or UHR criteria and thus were assumed to be at different points on the putative trajectory to psychosis. We compared them with 43 patients with a first episode of psychosis and to 49 help-seeking patient controls. All groups performed significantly below normative values. Both at-risk groups performed at intermediate levels between the first-episode (FE) group and normative values. The UHR group demonstrated intermediate performance between the FE and BS groups. Overall, auditory working memory, verbal fluency/processing speed, and declarative verbal memory were impaired the most. Our results suggest that cognitive impairments may still be modest in the early stages of the initial schizophrenia prodrome and thus support current efforts to intervene in the early course of impending schizophrenia because early intervention may prevent or delay the onset of frank psychosis and thus prevent further cognitive damage. PMID- 17412712 TI - Mutator pathways unleashed by epigenetic silencing in human cancer. AB - Human cancers exhibit genomic instability and an increased mutation rate due to underlying defects in DNA repair genes. Hypermethylation of CpG islands in gene promoter regions is an important mechanism of gene inactivation in cancer. Many cellular pathways, including DNA repair, are inactivated by this type of epigenetic lesion, resulting in mutator pathways. In this review, we discuss the adverse consequences suffered by a cell when DNA repair genes such as the DNA mismatch repair gene hMLH1, the DNA alkyl-repair gene O(6)-methylguanine-DNA methyltransferase, the familial breast cancer gene BRCA1 and the Werner syndrome gene WRN become epigenetically silenced in human cancer. PMID- 17412713 TI - Are public health physicians fading out of management? AB - BACKGROUND: Recent developments in health services in the local arena in Norway have challenged the theoretical and applied scientific basis for both public health medicine and management. During the 1990s although public health physicians in Norway increased in number, they worked less with public health, as well as public health management. The effects of these developments on public health management are largely unknown. We studied public health physicians' involvement in management and their self-reported managerial competence. METHODS: Cross-sectional study of physicians working in local public health medicine in all Norwegian municipalities, using a mail-back questionnaire. RESULTS: Public health physicians reduced their administrative tasks and evaluated their own managerial competence rather conservatively and somewhat lower in 1999 than in 1994. Many had supplementary training in management in addition to their medical education and specialty training. CONCLUSIONS: Public health physicians may be fading out of management. To address this there is a need for development of both public health management training programmes and provision of adequate resources for managerial activities. PMID- 17412714 TI - Barriers in accessing to tuberculosis care among non-residents in Shanghai: a descriptive study of delays in diagnosis. AB - OBJECTIVES: To describe accessibility to tuberculosis (TB) diagnosis in non resident TB patients in Shanghai, China, and to identify factors associated with delay in diagnosis. METHODS: A face-to-face interview of 222 newly diagnosed, non resident TB patients registered in two districts of Shanghai: Changning District and Putuo District, was conducted using a structured questionnaire. RESULTS: Among the 222 non-resident TB patients, median patient's delay was 21 days and median doctor's delay was 8 days. The duration of doctor's delay was significantly longer in Changning District than Putuo District (13 vs. 5 days, P < 0.001). One-fourth of the subjects had a patient's delay longer than 42 days and a doctor's delay longer than 15 days. Logistic regression model shows that patients at lower income level, and who did not have haemoptysis symptom were more likely to have longer patient's delay. Patients who registered in Changning were more likely to have a longer doctor's delay. The proportion of diagnosis or consideration as suspected TB for referral was significantly higher in hospitals than non-hospitals. CONCLUSION: The results of this study indicate that patient- and doctor-related factors contribute significantly to delays in the diagnosis of non-resident TB patients in Chinese cities. Non-resident's poor economic status, clinical status, complexities in referral and diagnostic procedure at different districts accounted for delayed TB care-seeking and diagnosis. PMID- 17412715 TI - Prescription of benzodiazepines in general practice in the context of a man-made disaster: a longitudinal study. AB - BACKGROUND: Mental health problems associated with benzodiazepine treatment are often highly prevalent in the aftermath of disasters. Nevertheless, not much is known about benzodiazepine use after disasters. Considering the negative effects associated with prolonged use and the adverse effects of benzodiazepines on recovery of patients with acute stress, the aim of the present study was to explore benzodiazepine use in the context of the Enschede fireworks disaster of 13 May 2000. METHODS: A longitudinal study using electronic medical records of general practitioners. Subjects were patients aged 16 years and older, registered at one of the practices between 1999 and 2003 (1541 victims and 5370 references). Pre- and post-disaster data were available on benzodiazepine prescriptions, healthcare utilization and sociodemographic characteristics. Benzodiazepine use was defined using different criteria (e.g. any use, daily use, chronic use). Data were analysed using multivariate multilevel logistic regression analyses. RESULTS: Compared with patients from a reference group, disaster victims were at increased risk of becoming an incident benzodiazepine user after the disaster. Benzodiazepine use also had a different time course among victims compared with references. However, daily or prolonged use of benzodiazepines was not often observed and did not show dramatic deviations among disaster victims compared with references. CONCLUSION: There is no convincing evidence that general practitioners systematically deviated from clinical guidelines for benzodiazepines, which generally advocate their short time application. PMID- 17412716 TI - Comparing MSM in the southeastern United States who participated in an HIV prevention chat room-based outreach intervention and those who did not: how different are the baseline HIV-risk profiles? AB - Chat room-based human immunodeficiency virus (HIV) prevention interventions are being implemented to reduce the risk of HIV exposure, infection and reinfection among men who have sex with men (MSM). However, little is known about how participants in chat room-based prevention interventions differ from their online non-participating peers. This analysis compared the baseline risk profiles of participants in an HIV prevention intervention ('active recruitment') to their chat room peers who did not participate in the intervention ('passive recruitment'). Data were collected using an online brief risk assessment from MSM (N = 448) who were recruited within Internet chat rooms. Mean age was 30 years. Half self-identified as Black or African American, 29% as White and 64% as gay. Compared with participants, non-participants were more likely to report: spending higher mean number of hours in online chat rooms; using condoms inconsistently during anal intercourse with a man met online during the past 3 months; having had an sexually transmitted disease; being HIV seropositive; using methamphetamines during the past 30 days and using drugs to enhance sexual satisfaction during the past 30 days. Although risk among MSM who use chat rooms remains high, those at greater risk may be those who are less likely to engage in online HIV prevention interventions. PMID- 17412717 TI - An Internet-based diabetes self-care intervention tailored to self-efficacy. AB - The main objective of this study was to assess whether self-efficacy (SE) could function as a moderator of the effect of a tailored Internet-based intervention aimed at increasing self-reported diabetes self-care behaviours. In a two-group, 1-month interval pre-test-post-test randomized controlled trial, participants (N = 64) were assigned at random to either a group that received an intervention on the area of self-care (blood glucose monitoring, diet management or physical activity) for which the reported SE was lowest (LSE group) or to a group that received an intervention on area of self-care for which the reported SE was highest (HSE group). Improvements in self-care were observed for both groups, but the HSE group improved more. Self-care also increased for those areas that the intervention did not target. Furthermore, SE levels decreased from baseline to follow-up. This study suggests that SE can function as a moderator in a behavioural intervention for diabetes self-care, and hence that initial level of SE provides relevant information for tailoring such interventions. PMID- 17412718 TI - Factors associated with the duration of exclusive breast-feeding in asthmatic families. AB - This study was part of a randomized controlled trial in which verbal and written advice about exclusive breast-feeding for 6 months was provided to Dutch women expecting a child with a high risk of developing asthmatic traits. Eighty-nine women completed a theory-based self-report questionnaire between the third and sixth months of pregnancy, which served as the baseline measurement. The aim of this study was to examine the factors that influence the duration of exclusive breast-feeding. Cox multiple regression analysis showed a positive significant association between the duration of exclusive breast-feeding and the mother's breast-feeding knowledge (P < 0.01), her intended hours of work per week after maternity leave (P < 0.01) and her age (P or =3 log(10) initial inocula reduction) against S. pneumoniae and S. aureus, regardless of the strain tested or the presence of albumin or human serum (that slightly delayed bactericidal activity). Against vancomycin susceptible or -resistant enterococci, daptomycin exhibited rapid bactericidal activity, delayed to 8 and 24 h, respectively, by human albumin. Vancomycin exhibited much slower bactericidal activity against MDRSP and methicillin susceptible or -resistant S. aureus, but was never bactericidal against MRSA-hVI and vancomycin-susceptible or -resistant E. faecium. CONCLUSIONS: Daptomycin exhibited rapid bactericidal activity against the strains of the three Gram positive species tested, regardless of resistance phenotype or the presence of physiological concentrations of albumin. PMID- 17412726 TI - Susceptibility of Pseudomonas aeruginosa to antimicrobials: a 2004 French multicentre hospital study. AB - OBJECTIVES: Pseudomonas aeruginosa is a major causative agent of hospital infections. The purpose of this study was to determine the antibiotic susceptibility of P. aeruginosa in a French multicentre study and to investigate the mechanisms of beta-lactam resistance. METHODS: Four hundred and fifty non repetitive strains of P. aeruginosa were collected in 15 French university hospitals in 2004. MICs of antibiotics were measured by agar dilution methods. For all the strains with MICs of ticarcillin >16 mg/L, detection and identification of the beta-lactamases, quantitative determination of cephalosporinase and overproduction of the MexAB-OprM efflux pump were evaluated. RESULTS: The percentages of susceptible isolates were as follows: ticarcillin, 62%; ticarcillin + clavulanic acid, 61%; piperacillin, 78%; piperacillin + tazobactam, 80% (MICs 0.4), compared with 0% of patients receiving etanercept. MRI analyses were inconclusive. Both drugs were well tolerated; 54% of infliximab treated patients and 50% of etanercept-treated patients reported adverse events. CONCLUSIONS: In this exploratory, open-label trial (with single-blind evaluator), patients were randomised to continue with etanercept or switch to infliximab. The small sample size of this hypothesis-generating study was underpowered to show statistical differences between groups. There was a numerical trend favouring patients who switched to infliximab, therefore warranting further study with a more rigorous design. PMID- 17412738 TI - B cell depletion therapy in systemic lupus erythematosus: long-term follow-up and predictors of response. AB - OBJECTIVES: To describe the long-term clinical outcome and safety profile of B cell depletion therapy (BCDT) in patients with systemic lupus erythematosus (SLE). It was also determined whether baseline parameters can predict the likelihood of disease flare. METHODS: 32 patients with refractory SLE were treated with BCDT using a combination protocol (rituximab and cyclo-phosphamide). Patients were assessed with the British Isles Lupus Assessment Group (BILAG) activity index, and baseline serology was measured. Flare was defined as a new BILAG 'A' or two new subsequent 'B's in any organ system. RESULTS: Of the 32 patients, 12 have remained well after one cycle of BCDT (median follow-up 39 months). BCDT was followed by a decrease of median global BILAG scores from 13 to 5 at 6 months (p = 0.006). Baseline anti-extractable nuclear antigen (ENA) was the only identified independent predictor of flare post-BCDT (p = 0.034, odds ratio = 8, 95% CI 1.2 to 55) from multivariable analysis. Patients with low baseline serum C3 had a shorter time to flare post-BCDT (p = 0.008). Four serious adverse events were observed. CONCLUSION: Autoantibody profiling may help identify patients who will have a more sustained response. Although the long-term safety profile of BCDT is favourable, ongoing vigilance is recommended. PMID- 17412739 TI - Asthma and airways collapse in two heritable disorders of connective tissue. AB - OBJECTIVES: This study investigated the clinical impression that there was an increased prevalence of respiratory disorders in both the Hypermobility Syndrome (HMS)/Benign Joint Hypermobility Syndrome (BJHS) and Ehlers-Danlos Syndrome (EDS), compared with the normal population. METHODS: A questionnaire was distributed to 509 subjects (221 healthy controls, 126 HMS, 162 EDS) who documented respiratory symptoms and previously diagnosed respiratory and atopic disorders. A subgroup of 157 responders underwent full clinical and serological assessments, and 57 subjects were assessed physiologically. RESULTS: A significant increase in the frequency of a wide range of respiratory symptoms and reduced exercise tolerance was observed in subjects with both HMS and EDS compared with controls. In particular, there was an increased prevalence of asthmatic symptoms (HMS: OR 2.4, 95% CI 1.4-4.1, p = 0.002; EDS: OR 3.1, 95% CI 1.8-5.2, p<0.001) and atopy (HMS: OR 2.7, 95% CI 1.6-4.5, p<0.001; EDS: OR 2.6, 95% CI 1.6-4.4, p<0.001), which was subsequently confirmed by clinical assessment. Pulmonary physiological studies revealed increased lung volumes, impaired gas exchange and an increased tendency of both the lower and upper airways to collapse. CONCLUSIONS: We have demonstrated, for the first time, that individuals with HMS/BJHS and EDS have respiratory symptoms in association with various pulmonary physiological abnormalities. The increased prevalence of asthma may be due to linkage disequilibrium between the genes causing these conditions or a function of the connective tissue defect itself. In the non-asthmatic population, changes in the mechanical properties of the bronchial airways and lung parenchyma may underlie the observed increased tendency of the airways to collapse. PMID- 17412740 TI - QUEST-RA: quantitative clinical assessment of patients with rheumatoid arthritis seen in standard rheumatology care in 15 countries. AB - OBJECTIVE: To conduct a cross-sectional review of non-selected consecutive outpatients with rheumatoid arthritis (RA) as part of standard clinical care in 15 countries for an overview of the characteristics of patients with RA. METHODS: The review included current disease activity using data from clinical assessment and a patient self-report questionnaire, which was translated into each language. Data on demographic, disease and treatment-related variables were collected and analysed using descriptive statistics. Variation in disease activity on DAS28 (disease activity score on 28-joint count) within and between countries was graphically analysed. A median regression model was applied to analyse differences in disease activity between countries. RESULTS: Between January 2005 and October 2006, the QUEST-RA (Quantitative Patient Questionnaires in Standard Monitoring of Patients with Rheumatoid Arthritis) project included 4363 patients from 48 sites in 15 countries; 78% were female, >90% Caucasian, mean age was 57 years and mean disease duration was 11.5 years. More than 80% of patients had been treated with methotrexate in all but three countries. Overall, patients had an active disease with a median DAS28 of 4.0, with a significant variation between countries (p<0.001). Among 42 sites with >50 patients included, low disease activity of DAS28 50% of patients had high disease activity of DAS28 >5.1. CONCLUSIONS: This international multicentre cross-sectional database provides an overview of clinical status and treatments of patients with RA in standard clinical care in 2005-6 including countries that are infrequently involved in clinical research projects. PMID- 17412741 TI - Cardiovascular outcomes in high risk patients with osteoarthritis treated with ibuprofen, naproxen or lumiracoxib. AB - BACKGROUND: Evidence suggests that both selective cyclooxygenase (COX)-2 inhibitors and non-selective non-steroidal anti-inflammatory drugs (NSAIDs) increase the risk of cardiovascular events. However, evidence from prospective studies of currently available COX-2 inhibitors and non-selective NSAIDs is lacking in patients at high cardiovascular risk who are taking aspirin. OBJECTIVE: To determine the cardiovascular outcomes in high risk patients with osteoarthritis treated with ibuprofen, naproxen or lumiracoxib. METHODS: The Therapeutic Arthritis Research and Gastrointestinal Event Trial (TARGET) of 18 325 patients with osteoarthritis comprised two parallel substudies, comparing lumiracoxib (COX-2 inhibitor) with either ibuprofen or naproxen. A post hoc analysis by baseline cardiovascular risk, treatment assignment, and low-dose aspirin use was performed. The primary composite end point was cardiovascular mortality, non-fatal myocardial infarction, and stroke at 1 year; a secondary end point was the development of congestive heart failure (CHF). RESULTS: In high risk patients among aspirin users, patients in the ibuprofen substudy had more primary events with ibuprofen than lumiracoxib (2.14% vs 0.25%, p = 0.038), whereas in the naproxen substudy rates were similar for naproxen and lumiracoxib (1.58% vs 1.48%, p = 0.899). High risk patients not taking aspirin had fewer primary events with naproxen than with lumiracoxib (0% vs 1.57%, p = 0.027), but not for ibuprofen versus lumiracoxib (0.92% vs 0.80%, p = 0.920). Overall, CHF developed more often with ibuprofen than lumiracoxib (1.28% vs 0.14%; p = 0.031), whereas no difference existed between naproxen and lumiracoxib. CONCLUSIONS: These data suggest that ibuprofen may confer an increased risk of thrombotic and CHF events relative to lumiracoxib among aspirin users at high cardiovascular risk. The study indicates that naproxen may be associated with lower risk relative to lumiracoxib among non-aspirin users. This study is subject to inherent limitations, and therefore should be interpreted as a hypothesis generating study. PMID- 17412742 TI - Priority setting using multiple criteria: should a lung health programme be implemented in Nepal? AB - OBJECTIVES: To identify and weigh the various criteria for priority setting, and to assess whether a recently evaluated lung health programme in Nepal should be considered a priority in that country. METHODS: Through a discrete choice experiment with 66 respondents in Nepal, the relative importance of several criteria for priority setting was determined. Subsequently, a set of interventions, including the lung health programme, was rank ordered on the basis of their overall performance on those criteria. RESULTS: Priority interventions are those that target severe diseases, many beneficiaries and people of middle age, have large individual health benefits, lead to poverty reduction and are very cost-effective. Certain interventions in tuberculosis control rank highest. The lung health programme ranks 13th out of 34 interventions. CONCLUSION: This explorative analysis suggests that the lung health programme is among the priorities in Nepal when taking into account a range of relevant criteria for priority setting. The multi-criteria approach can be an important step forward to rational priority setting in developing countries. PMID- 17412743 TI - Stress in mothers of young children with eczema. AB - OBJECTIVE: To assess parental stress levels of mothers of children less than 6 years old with eczema and compare these levels with those reported for other chronic childhood illnesses. METHODS: Mothers were recruited from hospital-based out-patient clinics (55%) or while their child was an in-patient (45%) for management of eczema. Maternal stress was measured utilising the Parenting Stress Index-Long Form (PSI) in 33 mothers. The severity of the eczema at the time of interview was documented by the Eczema Area and Severity Index (EASI) score and the Investigators' Global Assessment (IGA) score. RESULTS: The children with eczema had a mean age of 2.8 years. Mothers of children aged 5 years or less with eczema exhibited significantly higher total stress scores (mean PSI 259.6, 95% CI 244.9 to 274.3) as compared to mothers of normal children (PSI 222.8, 95% CI 221.4 to 224.2) and children with other chronic disorders such as insulin dependent diabetes (PSI 218.1, 95% CI 204.7 to 231.6) and profound deafness (PSI 221.7, 95% CI 206.4 to 237.0). Stress scores in the parental domain (138.2, 95% CI 128.9 to 147.6) did not differ significantly from the scores of parents of children with severe disabilities such as those requiring home enteral feeding (135.2, 95% CI 129.3 to 141.1) and those with Rett syndrome (132.8, 95% CI 125.0 to 140.6). CONCLUSIONS: Moderate to severe childhood eczema should be regarded as a significant illness in which maternal stress is equivalent to that associated with the care of children with severe developmental and physical problems. PMID- 17412744 TI - The contribution of single antigen measles, mumps and rubella vaccines to immunity to these infections in England and Wales. AB - OBJECTIVE: To obtain information on the use of single antigen measles, mumps and rubella vaccines to improve estimates of population immunity and help predict outbreaks. DESIGN: We requested information from providers of single antigen vaccines and from the Medicine and Healthcare products Regulatory Agency on requests for importation of single antigen measles and mumps vaccines. SETTING: England and Wales. MAIN OUTCOME MEASURES: Number of doses of single measles, mumps and rubella vaccine, by age of child (in months), year given and area of residence, and number of children who have received all three single vaccinations. RESULTS: Of 27 providers identified, 13 held single site clinics: nine were individual general practitioners and five held clinics at multiple sites. Data were received from 9/27 (33%) providers operating 40/74 (54%) clinic sites. We received information on 60 768 vaccinations administered by single vaccine providers and 269 917 doses requested for importation. For children born in 2001/2002, the minimum estimates for the proportion who received single measles vaccine are 1.7% in 2001 and 2.1% in 2002, with a reasonable maximum estimate of 5.6% over the 2 years. For single mumps vaccine, the minimum estimates are 0.3% in 2001 and 0.02% in 2002, with a maximum estimate of 4.0%. CONCLUSION: The contribution of single vaccines to immunity is small in comparison to that of the combined measles, mumps and rubella vaccine (MMR). For recent birth cohorts this contribution could increase routine coverage for measles-containing vaccines by around 2%, still below the level of immunity required to sustain elimination. PMID- 17412745 TI - Facilitating and understanding the family's choice of injection device for growth hormone therapy by using conjoint analysis. AB - BACKGROUND: Conjoint analysis involves the measurement of consumer preferences between choice alternatives. AIMS AND OBJECTIVES: To investigate the use of conjoint analysis in facilitating and understanding choice of growth hormone injection devices. METHOD AND SUBJECTS: 56 patients and their parents participated in an electronic, computer-based interview. The interview took a median time of 18 min (range 12-30) and allowed an immediate matching of injection devices to the family's preferences. RESULTS: Amongst the key drivers of choice, lack of bruising was rated highest and designated an index of 100. Compared to this, the remaining attributes in order of desirability were: auto injector (98), lack of pain (93), lightweight (88), silent (82), ready-mixed (77), ease of holding (69), telephone helpline (66), needle-free (62), small size (60), nurse support (47), hidden needle (45), stored in fridge (13) and home delivery (6). Out of the 17 families who had already chosen a device previously by discussion with the clinic nurse, the computer model placed their device either as first or second out of seven devices tested. CONCLUSION: Adaptive or interactive conjoint analysis applied at the patient level can facilitate the choice-making process whilst providing an insight into the relative importance of the key features that influence choice. PMID- 17412746 TI - Does intrauterine growth restriction affect quality of life in adulthood? AB - OBJECTIVE: To compare health-related quality of life in 50-year-old adults who were born at term (> or =37 to 42 weeks' gestation) with intrauterine growth restriction (IUGR; birth weight <10th centile) and a group born at term without IUGR (> or =10th centile). DESIGN: Case control study. SETTING: A large regional maternity hospital in Northern Ireland. SUBJECTS: 235 adults who were born between 1954 and 1956 in the Royal Maternity Hospital, Belfast. 111 subjects born with IUGR and 124 controls with normal birth weight for gestation were compared. MAIN OUTCOME MEASURE: Health-related quality of life in adulthood was assessed using the Short Form-36 Health Survey (SF-36). RESULTS: The two groups reported similar health-related quality of life on each of the eight dimensions of the SF 36 and there were no significant differences between them. Adjusting for potential confounding variables did not alter this conclusion. CONCLUSIONS: The similarity of SF-36 scores indicated that those born with IUGR did not perceive this to adversely influence health-related quality of life at 50 years of age compared with those with normal birth weight. PMID- 17412747 TI - Pattern of lithium exposure predicts poisoning severity: evaluation of referrals to a regional poisons unit. AB - BACKGROUND: Lithium toxicity may result in severe clinical features. There is on going uncertainty about the significance of serum lithium concentrations in patients with lithium toxicity. AIM: To examine potential relationships between stated quantity of lithium ingested, serum lithium concentrations, and poisoning severity among patients referred to a regional poisons centre. METHODS: Prospective evaluation of enquiries to the Scottish Poisons Information Bureau about lithium toxicity between 2000-2005 inclusive. RESULTS: There were 172 enquiries, relating to acute ingestion (n = 101), acute-on-therapeutic ingestion (n = 38), or chronic poisoning (n = 33). Poisoning severity was moderate or severe in 9.9%, 26.3% (p < 0.05 vs. acute) and 54.5% (p < 0.005 vs. acute) of each group, respectively. Median (IQR) serum lithium concentrations in each group were: 2.4 (1.7-3.3) mmol/l, 2.1 (1.4-3.8) mmol/l, and 2.3 (1.9-3.3) mmol/l, respectively. The median stated quantities ingested in acute and acute-on therapeutic lithium exposure were 5000 mg (2000-11 050 mg) and 4000 mg (2400-8820 mg), respectively. DISCUSSION: Patients with acute-on-therapeutic and chronic poisoning are at greatest risk of severe toxicity. These differences cannot be explained by either the quantity of lithium ingested or serum lithium concentration alone. PMID- 17412748 TI - Car seat test for preterm infants: comparison with polysomnography. AB - OBJECTIVES: To monitor preterm infants in a cot and a car seat and compare an observed car seat trial with polysomnography (PSG). DESIGN: Non-randomised controlled trial. SETTING: Regional neonatal unit. PATIENTS: Preterm infants before discharge. INTERVENTIONS: Nap PSG respiratory and sleep variables were measured including gastro-oesophageal pH. Nurse observations included respiratory distress, apnoea measured by apnoea alarm, oxygen saturation and heart rate. Infants were studied supine in a cot and then in a car seat. Nursing observations were compared with PSG during the car seat trial only. Criteria for failure of the PSG and observed tests were predefined. MAIN OUTCOME MEASURES: Difference in respiratory instability between cot and car seat. Concurrence regarding failure of the car seat trial between nurse-observed data and PSG. RESULTS: 20 infants (median gestation 33 weeks (range 28-35 weeks; median postmenstrual age (PMA) at study 36.5 weeks (range 35-38 weeks)) were studied. There were sufficient car seat data on 18 infants for comparison. There were fewer central apnoeas and arousals in the cot than the car seat (p = 0.047 and p = 0.024, respectively). Airway obstruction was not more common in the car seat. Younger PMA at time of study predicted failure in both car seat (p = 0.022) and cot (p = 0.022). The nurse-observed test had low sensitivity for predicting PSG failure but more accurately predicted airway obstruction on PSG. CONCLUSIONS: Immature infants exhibit respiratory instability in cots and car seats. A car seat test does not accurately detect all adverse events during sleep in the seat. PMID- 17412749 TI - Methicillin-resistant Staphylococcus aureus infections among healthy full-term newborns. AB - OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) strains have emerged in the community, causing disease among healthy people lacking traditional risk factors for MRSA infection. This article describes an outbreak of MRSA among healthy full-term newborns. DESIGN: Cases were identified and corresponding medical information collected. Telephone interviews were conducted with mothers of cases and surveillance cultures from mothers and newborns were performed. MRSA isolates were genotyped. SETTING: Hospital in Chicago, Illinois, USA. PARTICIPANTS: Newborns, their mothers and hospital healthcare workers. INTERVENTION: Nursery infection control practices were enhanced. The MRSA colonised healthcare workers received intranasal mupirocin. MAIN OUTCOME: Within 4-23 days of birth, 11 newborns were identified with pustules, vesicles or blisters located on the head, groin, perineum, ears, legs, chin and trunk. All received antimicrobials and recovered without incident. RESULTS: None of 432 peripartum women, one of 399 newborns, and two of 135 healthcare workers were nasal MRSA carriers. Available isolates from six patients, two healthcare workers, and one from an MRSA-colonised newborn were similar by pulsed-field gel electrophoresis. Other than contact with the hospital, no common exposures of MRSA transmission were identified. CONCLUSIONS: MRSA strains that initially emerged in the community are now causing disease in healthcare settings. Providers should be aware that MRSA can cause skin infections among healthy newborns. Adherence to standard infection control practices is important to prevent transmission of MRSA in nurseries. PMID- 17412750 TI - Interaction between genioglossus and diaphragm responses to transcranial magnetic stimulation in awake humans. AB - The modulation of activity of the upper airway dilator and respiratory muscles plays a key role in the regulation of ventilation, but little is known about the link between their neuromuscular activation processes in vivo. This study investigated genioglossus and diaphragm responses to transcranial magnetic stimulation applied in different facilitatory conditions. The amplitude and latency of motor-evoked potential responses and the stimulation intensity threshold leading to a motor response (motor threshold) were recorded with stimulation applied at the vertex and anterolateral area in 13 awake normal subjects. Stimuli were applied during inspiration with and without resistance, during expiration with and without maximal tongue protrusion and during deep inspiration. In each stimulation location and condition, no diaphragmatic response was obtained without previous genioglossus activity (diaphragmatic and genioglossus responses latencies during expiration: 18.1 +/- 2.9 and 6.3 +/- 2.6 ms, respectively, mean +/- s.d., P < 0.01). Genioglossus motor-evoked potential amplitude, latency and motor threshold were significantly modified with tongue protrusion with a maximal effect observed for stimulation in the anterolateral area. Deep inspiration was associated with a significant facilitatory effect on both genioglossus and diaphragm motor responses. The facilitatory effects of respiratory and non-respiratory manoeuvres were also observed during focal stimulation where isolated genioglossus responses were observed. Genioglossus and diaphragm differed in their motor threshold both at baseline and following facilitatory manoeuvres. CONCLUSIONS: (1) transcranial magnetic stimulation induced genioglossus response systematically precedes that of diaphragm; (2) this sequence of activation is not modified by respiratory and non-respiratory manoeuvres; and (3) the genioglossus and diaphragm are differently influenced by these manoeuvres in terms of latency of the motor response and of motor threshold. PMID- 17412751 TI - Attack and defence in the gastric epithelium - a delicate balance. AB - The gastric epithelium is a complex structure formed into tubular branched gastric glands. The glands contain a wide variety of cell types concerned with the secretion of hydrochloric acid, proteases, mucus and a range of signalling molecules. All cell types originate from stem cells in the neck region of the gland, before migrating and differentiating to assume their characteristic positions and functions. Endocrine and local paracrine mediators are of crucial importance for maintaining structural and functional integrity of the epithelium, in the face of a hostile luminal environment. The first such mediator to be recognized, the hormone gastrin, was identified over a century ago and is now established as the major physiological stimulant of gastric acid secretion. Recent studies, including those using mice that overexpress or lack the gastrin gene, suggest a number of previously unrecognized roles for this hormone in the regulation of cellular proliferation, migration and differentiation. This review focuses on the identification of hitherto unsuspected gastrin-regulated genes and discusses the paracrine cascades that contribute to the maintenance of gastric epithelial architecture and secretory function. Helicobacter infection is also considered in cases where it shares targets and signalling mechanisms with gastrin. PMID- 17412752 TI - Independent counselling on embryo donation for infertility patients. PMID- 17412753 TI - Comparing highly purified hMG and rFSH in patients undergoing IVF. PMID- 17412754 TI - Signatures of recent positive selection at the ATP-binding cassette drug transporter superfamily gene loci. AB - Members of the ATP-binding cassette (ABC) superfamily of transporters have been implicated as major players in drug response. Single nucleotide polymorphisms (SNPs) in the ABC transporter genes may account for variation in drug response between individuals. Given the abundance of SNPs within the human genome, identification of functionally important SNPs is difficult. Here, we utilized signatures of recent positive selection (RPS) to identify SNPs in ABC genes that have potential functional significance by using the long-range-haplotype test to search for signatures of RPS at 18 ABC genes involved in drug transport. From the genotype data of these 18 ABC genes in four populations extracted from the HapMap database, at least one SNP in each of these genes displayed genomic signatures of RPS in at least one population. However, only 13 SNPs in 10 ABC genes from three populations retained statistical significance after Type I error reduction. The functional significance of six of these RPS SNPs, including those that failed multiple testing correction (MTC), has been reported previously. We experimentally confirmed a functional effect for two SNPs, including one that failed to show evidence of RPS after MTC. These observations suggest that Type I error reduction may inadvertently increase Type II error. Although the remaining positively selected SNPs have yet to be functionally validated, our study illustrates the feasibility of using this strategy to identify SNPs within 'adaptive' genes that may confer functional effect, prior to testing their roles in individual/population drug response variation or in complex disease susceptibility. PMID- 17412755 TI - Functional polymorphism in ABCA1 influences age of symptom onset in coronary artery disease patients. AB - ATP-binding-cassette-transporter-A1 (ABCA1) plays a pivotal role in intracellular cholesterol removal, exerting a protective effect against atherosclerosis. ABCA1 gene severe mutations underlie Tangier disease, a rare Mendelian disorder that can lead to premature coronary artery disease (CAD), with age of CAD onset being two decades earlier in mutant homozygotes and one decade earlier in heterozygotes than in mutation non-carriers. It is unknown whether common polymorphisms in ABCA1 could influence age of symptom onset of CAD in the general population. We examined common promoter and non-synonymous coding polymorphisms in relation to age of symptom onset in a group of CAD patients (n = 1164), and also carried out in vitro assays to test effects of the promoter variations on ABCA1 promoter transcriptional activity and effects of the coding variations on ABCA1 function in mediating cellular cholesterol efflux. Age of symptom onset was found to be associated with the promoter - 407G > C polymorphism, being 2.82 years higher in C allele homozygotes than in G allele homozygotes and intermediate in heterozygotes (61.54, 59.79 and 58.72 years, respectively; P = 0.002). In agreement, patients carrying ABCA1 haplotypes containing the -407C allele had higher age of symptom onset. Patients of the G/G or G/C genotype of the -407G > C polymorphism had significant coronary artery stenosis (>75%) at a younger age than those of the C/C genotype (P = 0.003). Reporter gene assays showed that ABCA1 haplotypes bearing the -407C allele had higher promoter activity than haplotypes with the -407G allele. Functional analyses of the coding polymorphisms showed an effect of the V825I substitution on ABCA1 function, with the 825I variant having higher activity in mediating cholesterol efflux than the wild-type (825V). A trend towards higher symptom onset age in 825I allele carriers was observed. The data indicate an influence of common ABCA1 functional polymorphisms on age of symptom onset in CAD patients. PMID- 17412756 TI - Duplication of the entire 22.9 Mb human chromosome 21 syntenic region on mouse chromosome 16 causes cardiovascular and gastrointestinal abnormalities. AB - Down syndrome is caused by a genomic imbalance of human chromosome 21 which is mainly observed as trisomy 21. The regions on human chromosome 21 are syntenically conserved in three regions on mouse chromosomes 10, 16 and 17. Ts65Dn mice, the most widely used model for Down syndrome, are trisomic for approximately 56.5% of the human chromosome 21 syntenic region on mouse chromosome 16. To generate a more complete trisomic mouse model of Down syndrome, we have established a 22.9 Mb duplication spanning the entire human chromosome 21 syntenic region on mouse chromosome 16 in mice using Cre/loxP-mediated long-range chromosome engineering. The presence of the intact duplication in mice was confirmed by fluorescent in situ hybridization and BAC-based array comparative genomic hybridization. The expression levels of the genes within the duplication interval reflect gene-dosage effects in the mutant mice. The cardiovascular and gastrointestinal phenotypes of the mouse model were similar to those of patients with Down syndrome. This new mouse model represents a powerful tool to further understand the molecular and cellular mechanisms of Down syndrome. PMID- 17412757 TI - The pathomechanism of filaminopathy: altered biochemical properties explain the cellular phenotype of a protein aggregation myopathy. AB - Myofibrillar myopathy (MFM) is a pathologically defined group of hereditary human muscle diseases, characterized by focal myofibrillar destruction and cytoplasmic aggregates that contain several Z-disc-related proteins. The previously reported MFM-associated mutation (8130G --> A; W2710X) in the filamin C gene (FLNC) leads to a partial disturbance of the secondary structure of the dimerization domain of filamin C, resulting in massive protein aggregation in skeletal muscle fibers of the patients. Here, we provide a thorough characterization of the biochemical, biophysical and cellular properties of the mutated filamin C polypeptide. Our experiments revealed that the mutant dimerization domain is less stable and more susceptible to proteolysis. As a consequence, it does not dimerize properly and forms aggregates in vitro. Furthermore, the expression of mutant filamin in cultured cells results in the formation of protein aggregates. The mutant filamin does not associate with wild type filamin. These findings are of great importance to explain the pathomechanism of this disease. PMID- 17412758 TI - Association between the T-381C polymorphism of the brain natriuretic peptide gene and risk of type 2 diabetes in human populations. AB - Brain natriuretic peptide (BNP/NPPB) is a member of the natriuretic family involved in the regulation of blood pressure and blood volume as well as lipolysis control in human fat cells. Thus BNP may play a role in energy metabolism and metabolic diseases. We therefore assessed the association between the BNP promoter T-381C polymorphism and risk of type 2 diabetes and metabolic and BNP expression traits in several population samples. In French population based samples (n = 3216), we found that individuals bearing the -381CC genotype had lower (P = 0.005) fasting glucose levels than -381TC or -381TT individuals. Moreover, the -381CC genotype was less frequent in individuals with type 2 diabetes (n = 280, 13.6%) or with impaired fasting glucose (n = 248, 12.9%) compared with normoglycaemic individuals (n = 2485, 17.8%). The adjusted odds ratio (OR) (95% CI) of type 2 diabetes for -381CC individuals was 0.69 (0.47 1.00), P = 0.05, when compared with -381T allele bearers. We replicated this association in four additional case-control studies for type 2 diabetes. The overall OR (95% CI) of type 2 diabetes was 0.85 (0.76-0.96), P = 0.008, (under a recessive model) (3593 cases and 6646 controls in total). We also found that the 381C allele was associated with higher plasma BNP concentrations (P = 0.015, n = 634) and higher BNP promoter activity in reporter gene assays. Collectively, these data suggest that relatively high BNP expression may protect against type 2 diabetes in humans. PMID- 17412759 TI - Mono- and double-mutant mouse models of Parkinson's disease display severe mitochondrial damage. AB - Mutations in the gene encoding alpha-synuclein (asyn) causes autosomal-dominant, in the parkin gene autosomal-recessive forms of Parkinson's disease (PD). The pathophysiology of PD is poorly understood, even though published evidence suggests a role for mitochondria in the pathogenesis. To gain insight into the influence of asyn and parkin on mitochondrial integrity and function, we have generated several mono-mutant mouse lines expressing doubly mutated human asyn (hm(2)asyn) under the control of two different promoters, or a targeted deletion of Parkin (Parkin-Exon3-knockout). Both mouse lines were crossed to generate the double-mutant. Here we compare the ultrastructure and functional properties of mitochondria in the substantia nigra (SN), the striatum, the cerebral cortex (Cx) and skeletal muscle of young (2-3 months) and aged (12-14 months) mono- and double-mutants mice. We observed severe genotype-, age- and region-dependent morphological alterations of mitochondria in neuronal somata. The number of structurally altered mitochondria was significantly increased in the SN of both double-mutants and in the Cx of one mono- and one double-mutant line. These alterations coincided with a reduced complex I capacity in the SN, but were neither accompanied by alterations in the number or the size of the mitochondria nor by leakage of cytochrome c, Smac/DIABLO or Omi/HtrA2. None of the transgenic animals developed any gross histopathological abnormalities or overt motor disabilities. Together our results provide compelling evidence that (i) both, asyn and parkin are relevant for mitochondrial integrity, (ii) the influence of these proteins on mitochondria are age- and tissue-specific and (iii) changes of mitochondrial morphology do not inevitably cause functional impairments. PMID- 17412760 TI - Epigenetic defects of hepatocellular carcinoma are already found in non neoplastic liver cells from patients with hereditary haemochromatosis. AB - Gene silencing through aberrant CpG island methylation is a frequent epigenetic defect in hepatocellular carcinoma (HCC). However, nothing is known as yet whether aberrant hypermethylation occurs already in non-neoplastic liver cells from patients with hereditary haemochromatosis who have a clearly elevated risk for developing HCC. Therefore, quantitative real-time PCR-based methylation analysis of six genes frequently hypermethylated in HCC (RASSF1A, cyclinD2, p16(INK4a), GSTpi1, SOCS-1, APC) was performed for liver biopsies from patients with hereditary haemochromatosis. For genotyping of the HFE gene restriction enzyme analysis and Pyrosequencing were used. Transcriptional repression of hypermethylated genes was assessed using real-time RT-PCR. Eighty-four percent of all samples with severe hepatic iron overload and a mutated HFE gene (but without HCC) had at least one gene hypermethylated. All six genes tested were affected by aberrant hypermethylation, albeit to a different extent: RASSF1A 55%, cyclinD2 45%, p16(INK4a) 32%, GSTpi1 10%, SOCS-1 6%, APC 8%. Concomitant transcriptional down-regulation was shown for RASSF1A, cyclinD2, GSTpi1 and SOCS-1. Biopsies from haemochromatosis patients showed significantly more aberrant hypermethylation than normal liver tissue or benign liver tumours (P < 0.001) and also to a higher degree. This effect is independent of patient age, cirrhosis or hepatitis infection. This is the first report demonstrating that longstanding severe iron overload is frequently associated with epigenetic defects characteristic of HCC, which reflects the increased risk of these lesions to progress to HCC. Thus, changes in DNA methylation patterns are an early event preceding morphological alterations of malignant transformation and represent promising targets for early detection. PMID- 17412761 TI - Local anaesthetic block of sodium channels: raising the barrier. PMID- 17412762 TI - Target-specific PIP(2) signalling: how might it work? AB - Phosphatidylinositol 4,5-bisphosphate (PIP(2))-mediated signalling is a new and rapidly developing area in the field of cellular signal transduction. With the extensive and growing list of PIP(2)-sensitive membrane proteins (many of which are ion channels and transporters) and multiple signals affecting plasma membrane PIP(2) levels, the question arises as to the cellular mechanisms that confer specificity to PIP(2)-mediated signalling. In this review we critically consider two major hypotheses for such possible mechanisms: (i) clustering of PIP(2) in membrane microdomains with restricted lateral diffusion, a hypothesis providing a mechanism for spatial segregation of PIP(2) signals and (ii) receptor-specific buffering of the global plasma membrane PIP(2) pool via Ca(2+)-mediated stimulation of PIP(2) synthesis or release, a concept allowing for receptor specific signalling with free lateral diffusion of PIP(2). We also discuss several other technical and conceptual intricacies of PIP(2)-mediated signalling. PMID- 17412763 TI - Regulation of KCNQ channels by manipulation of phosphoinositides. AB - Activation of phospholipase C (PLC) through G-protein-coupled receptors produces a large number of second messengers and regulates many physiological processes. Many membrane proteins including ion channels require the phosphoinositide phosphatidylinositol 4,5-bisphosphate (PIP(2)) to function. Activation of PLC can shut down their activity if it depletes the PIP(2) pool strongly. Such a mechanism accounts for the muscarinic suppression of current in KCNQ channels. We describe a variety of methods used to show that these channels require PIP(2) and that current in the channels is suppressed when receptor-activated PLC depletes PIP(2). The methods include observing translocation of lipid-sensitive protein domains, overexpression of enzymes of phosphoinositide metabolism, engineering these enzymes to move to the plasma membrane in response to a chemical signal, and direct chemical analysis of phospholipids. These approaches are general and can be used to test for PIP(2) requirements of other membrane proteins. PMID- 17412764 TI - Structural and functional basis of amino acid specificity in the invertebrate cotransporter KAAT1. AB - The substrate specificity of KAAT1, a Na+- and K+-dependent neutral amino acid cotransporter cloned from the larva of the invertebrate Manduca sexta and belonging to the SLC6A gene family has been investigated using electrophysiological and radiotracer methods. The specificity of KAAT1 was compared to that of CAATCH1, a strictly related transporter with different amino acid selectivity. Competition experiments between different substrates indicate that both transporters bind leucine more strongly than threonine and proline, the difference between KAAT1 and CAATCH1 residing in the incapacity of the latter to complete the transport cycle in presence of leucine. The behaviour of CAATCH1 is mimicked by the S308T mutant form of KAAT1, constructed on the basis of the atomic structure of a leucine-transporting bacterial member of the family, which indicates the participation of this residue in the leucine-binding site. The reverse mutation T308S in CAATCH1 conferred to this transporter the ability to transport leucine in presence of K+. These results may be interpreted by a kinetic scheme in which, in presence of Na+, the leucine-bound state of the transporter is relatively stable, while in presence of K+ and at negative potentials the progression of the leucine-bound form along the cycle is favoured. In this context serine 308 appears to be important in allowing the change to the inward-facing conformation of the transporter following substrate binding, rather than in determining the binding specificity. PMID- 17412765 TI - A hydrophobic element secures S4 voltage sensor in position in resting Shaker K+ channels. AB - The S4 transmembrane alpha-helix in voltage-gated channels contains several regularly spaced basic amino acid residues that could be protonated and moved across the membrane electric field in response to membrane potential changes. The translocation of the charge-carrying S4 transduces membrane voltage to gating conformational changes of the channel, but how it is positioned and moved with respect to membrane lipid remains controversial. We found that hydrophilic and especially arginine and lysine substitution for L361 at the external end of S4 causes a large negative shift with shallowed slope of both activation and inactivation curves in Shaker K+ channels. Also, the macroscopic kinetics of activation and inactivation become much faster and barely voltage dependent, especially in the L361R mutant channel. These steady-state and kinetic data suggest that the replacement of one single hydrophobic residue, leucine, with arginine may profoundly destabilize the resting conformation of S4, which therefore takes a partially extruded position (partly activated position) at resting potentials (e.g. -120 mV). Consistently, the L361R point mutation gives rise to an extracellularly exposed R365C that is readily modified by external hydrophilic sulfhydryl-specific agents in the resting channel. Moreover, the extruded S4 in the L361R mutant channel could be retracted by strong hyperpolarizing potentials ( approximately -180 mV), from which the mutant channel is gated with slower kinetics but evidently stronger voltage dependence. We conclude that hydrophobic interaction involving a highly conserved residue at the top of S4 is crucial for properly securing the gating voltage sensor in the resting position and thus appropriate gating control of the voltage-gated channels. PMID- 17412766 TI - Astrocytic control of synaptic NMDA receptors. AB - Astrocytes express a wide range of G-protein coupled receptors that trigger release of intracellular Ca2+, including P2Y, bradykinin and protease activated receptors (PARs). By using the highly sensitive sniffer-patch technique, we demonstrate that the activation of P2Y receptors, bradykinin receptors and protease activated receptors all stimulate glutamate release from cultured or acutely dissociated astrocytes. Of these receptors, we have utilized PAR1 as a model system because of favourable pharmacological and molecular tools, its prominent expression in astrocytes and its high relevance to neuropathological processes. Astrocytic PAR1-mediated glutamate release in vitro is Ca2+ dependent and activates NMDA receptors on adjacent neurones in culture. Activation of astrocytic PAR1 in hippocampal slices induces an APV-sensitive inward current in CA1 neurones and causes APV-sensitive neuronal depolarization in CA1 neurones, consistent with release of glutamate from astrocytes. PAR1 activation enhances the NMDA receptor-mediated component of synaptic miniature EPSCs, evoked EPSCs and evoked EPSPs in a Mg2+-dependent manner, which may reflect spine head depolarization and consequent reduction of NMDA receptor Mg2+ block during subsequent synaptic currents. The release of glutamate from astrocytes following PAR1 activation may also lead to glutamate occupancy of some perisynaptic NMDA receptors, which pass current following relief of tonic Mg2+ block during synaptic depolarization. These results suggest that astrocytic G-protein coupled receptors that increase intracellular Ca2+ can tune synaptic NMDA receptor responses. PMID- 17412767 TI - Mechanisms of cardiac potassium channel trafficking. AB - The regulation of ion channels involves more than just modulation of their synthesis and kinetics, as controls on their trafficking and localization are also important. Although the body of knowledge is fairly large, the entire trafficking pathway is not known for any one channel. This review summarizes current knowledge on the trafficking of potassium channels that are expressed in the heart. Our knowledge of channel assembly, trafficking through the Golgi apparatus and on to the surface is covered, as are controls on channel surface retention and endocytosis. PMID- 17412768 TI - Role of CNT3 in the transepithelial flux of nucleosides and nucleoside-derived drugs. AB - We examined the role of the concentrative nucleoside transporter CNT3 in the establishment of a transepithelial flux of natural nucleosides and their pharmacologically active derivatives in renal epithelial cell lines. Murine PCT cells grown on a transwell dish showed endogenous CNT3 activity at their apical membrane that was responsible for the sodium-dependent transepithelial flux of both purine and pyrimidine nucleosides. hCNT3 was also identified in human kidney and its role in the transport of nucleosides was tested. To this end, MDCK cells, lacking endogenous CNT3 activity, were genetically engineered to express the human orthologue of CNT3 (hCNT3-MDCK cells). In these cells, hCNT3 was inserted into the apical membrane, thus generating, as for PCT cells, a transepithelial flux of both nucleosides and nucleoside-derived drugs. Apical-to-basolateral transepithelial flux was present in all cells expressing a functional CNT3 transporter and was significantly higher than that found either in PCT cells in absence of sodium or in mock-transfected MDCK cells. Nevertheless in all cases a significant amount of the transported nucleoside was retained and transformed inside cells. However release to the opposite compartment was CNT3 dependent, not only in terms of absolute flux (much higher when an apical CNT3 transporter was active) but also regarding metabolic transformations of the apically absorbed nucleosides. These results underline a critical role of CNT3 in the renal reabsorption of nucleosides and their derivatives as well as in their intracellular metabolism. PMID- 17412769 TI - AMPA receptors associated with zebrafish Mauthner cells switch subunits during development. AB - Glutamate AMPA receptors (AMPARs) are major excitatory receptors in the vertebrate CNS. In many biological systems there is a developmental speeding in AMPAR kinetics, which occurs either because of a switch in AMPAR subunits or a change in synaptic morphology. We studied the development of AMPAR-mediated miniature excitatory postsynaptic currents (AMPAR-mEPSCs) in zebrafish Mauthner cells (M-cells) to determine the reasons underlying the speeding of AMPA mEPSCs in this preparation. We recorded AMPAR-mEPSCs in zebrafish ranging in age from 33 h postfertilization (hpf) to 72 hpf. We found that the glutamate waveform in the synaptic cleft did not change during development, suggesting that synaptic morphology played little role in shaping the mEPSC. The current-voltage (I-V) relationship was linear at 33 hpf and outwardly rectified in older animals, while AMPAR decay kinetics were slower at positive potentials, compared with negative potentials. The relative change in tau with depolarization was found to be greater at 48 hpf than at 33 hpf. AMPARs in 33 hpf fish had a conductance of approximately 9 pS, and in older fish approximately 15 pS. Finally, the desensitization blocker, cyclothiazide, increased tau by approximately 4-fold in 48 hpf preparations, but only 1.5-fold in 33 hpf fish. These results are consistent with the hypothesis that the major mechanism underlying the developmental speeding in AMPAR kinetics in zebrafish CNS is a switch in receptor subunits. To our knowledge this is the first study to suggest that AMPARs change subunits during development in fish. PMID- 17412770 TI - Linking lateral interactions in flicker perception to lateral geniculate nucleus cell responses. AB - The perception of flicker strength in a circular stimulus can be changed by altering the relative temporal phase of a simultaneously flickering surrounding annulus: perceived flicker is weak when the two stimuli are modulated in-phase and strong when the two are modulated in counter-phase. Previously, we found that responses of single neurons in the monkey lateral geniculate nucleus (LGN) to such stimuli resemble the psychophysical data. On the basis of the resemblance in data, it was proposed that the physiological basis for the flicker perception may be present as proximal as the LGN. To strengthen this hypothesis, we simulated the response of an array of LGN neurons, the receptive fields (RFs) of which are covered by the stimulus. The simulations were based upon single-cell recordings in the LGN of anaesthetized marmosets (Callithrix jacchus) using the same stimuli as previously. The measurements were repeated for different spatial displacement between the stimulus and the RF. The responses depended upon the spatial displacement and the relative phase between centre and surround stimuli. The neuronal responses can be adequately described by a difference-of-Gaussians (DOG) model with a time delay in the RF surround. The model responses at different displacements can be considered to be identical to the output of an array of ideal and identical LGN cells with different RF locations. To be able to describe physiological and psychophysical data, obtained at different stimulus contrasts, it was necessary to consider previously described non-linear interactions between the RF centres and surrounds. We applied a spatial peak-to-trough detector with a subsequent saturation and threshold to simulate a simple cortical decision mechanism. The output of this peak-to-trough detector could adequately describe the psychophysical data. PMID- 17412771 TI - Cerebrovascular reactivity to hypercapnia is unimpaired in breath-hold divers. AB - Hypercapnic cerebrovascular reactivity is decreased in obstructive sleep apnoea and congestive heart disease perhaps as a result of repeated apnoeas. To test the hypothesis that repeated apnoeas blunt cerebrovascular reactivity to hypercapnia, we studied breath hold divers and determined cerebrovascular reactivity by measuring changes in middle cerebral artery velocity (MCAV, cm s(-1)) per mmHg change in end-tidal partial pressure of CO2(PET,CO2 ) in response to two hyperoxic hypercapnia rebreathing manoeuvres (modified Read protocol) in elite breath-hold divers (BHD, n=7) and non-divers (ND, n=7). In addition, ventilation and central (beat-to-beat stroke volume measurement with Modelflow technique) haemodynamics were determined. Ventilatory responses to hypercapnia were blunted in BHD versus ND largely due to lower breathing frequency. Cerebrovascular reactivity did not differ between groups (3.7 +/- 1.4 versus 3.4 +/- 1.3% mmHg( 1) in BHD and ND, respectively; P=0.90) and the same was found for cerebral vascular resistance and MCAV recovery to baseline after termination of the CO2 challenge. Cardiovascular parameters were not changed significantly during rebreathing in either group, except for a small increase in mean arterial pressure for both groups. Our findings indicate that the regulation of the cerebral circulation in response to hypercapnia is intact in elite breath-hold divers, potentially as a protective mechanism against the chronic intermittent cerebral hypoxia and/or hypercapnia that occurs during breath-hold diving. These data also suggest that factors other than repeated apnoeas contribute to the blunting of cerebrovascular reactivity in conditions like sleep apnoea. PMID- 17412772 TI - Characteristics of the response of the iliac artery to wall shear stress in the anaesthetized pig. AB - The functional significance of shear stress-induced vasodilatation in large conduit arteries is unclear since changes in the diameter have little effect on the resistance to blood flow. However, changes in diameter have a relatively large effect on wall shear stress which suggests that the function of flow mediated dilatation is to reduce wall shear stress. The mean and pulsatile components of shear stress vary widely throughout the arterial system and areas of low mean and high amplitude of wall shear stress are prone to the development of atheroma. In this study, using an in vivo model with the ability to control flow rate and amplitude of flow independently, we investigated the characteristics of the response of the iliac artery to variations in both the mean and amplitude of wall shear stress. The results of this study confirm that increases in mean wall shear stress are an important stimulus for the release of nitric oxide by the endothelium as indicated by changes in arterial diameter and show for the first time, in vivo, that increases in the amplitude of the pulsatile component of shear stress have a small but significant inhibitory effect on this response. A negative feedback mechanism was identified whereby increases in shear stress brought about by increases in blood flow are reduced by the release of nitric oxide from the endothelium causing dilatation of the artery, thus decreasing the stimulus to cell adhesion and, through a direct action of nitric oxide, inhibiting the process of cell adhesion. The results also provide an explanation for the uneven distribution of atheroma throughout the arterial system, which is related to the ratio of pulsatile to mean shear stress and consequent variability in the production of NO. PMID- 17412773 TI - Effects of repeated prenatal glucocorticoid exposure on long-term potentiation in the juvenile guinea-pig hippocampus. AB - Synthetic glucocorticoids (sGCs) are routinely used to treat women at risk of preterm labour to promote fetal lung maturation. There is now strong evidence that exposure to excess glucocorticoid during periods of rapid brain development has permanent consequences for endocrine function and behaviour in the offspring. Prenatal exposure to sGC alters the expression of N-methyl-D-aspartate receptor (NMDA-R) subunits in the fetal and neonatal hippocampus. Given the integral role of the NMDA-R in synaptic plasticity, we hypothesized that prenatal sGC exposure will have effects on hippocampal long-term potentiation (LTP) after birth. Further, this may occur in either the presence or absence of elevated cortisol concentrations, in vitro. Pregnant guinea-pigs were injected with betamethasone (Beta, 1 mg kg(-1)) or vehicle on gestational days (gd) 40, 41, 50, 51, 60 and 61 (term approximately 70 days), a regimen comparable to that given to pregnant women. On postnatal day 21, LTP was examined at Schaffer collateral synapses in the CA1 region of hippocampal slices prepared from juvenile animals exposed to betamethasone or vehicle, in utero. Subsequently, the acute glucocorticoid receptor (GR)- and mineralocorticoid receptor (MR)-dependent effects of cortisol (0.1-10 microM; bath applied 30 min before LTP induction) were examined. There was no effect of prenatal sGC treatment on LTP under basal conditions. The application of 10 microM cortisol depressed excitatory synaptic transmission in all treatment groups regardless of sex. Similarly, LTP was depressed by 10 microM cortisol in all groups, with the exception of Beta-exposed females, in which LTP was unaltered. Hippocampal MR and GR protein levels were increased in Beta exposed females, but not in any other prenatal treatment group. This study reveals sex-specific effects of prenatal exposure to sGC on LTP in the presence of elevated cortisol, a situation that would occur in vivo during stress. PMID- 17412774 TI - Impairment of human proprioception by high-frequency cutaneous vibration. AB - These experiments assessed whether the impairment in proprioceptive acuity in the hand during 'interfering' cutaneous stimulation could be caused by inputs from Pacinian corpuscles. The ability to detect passive movements at the proximal interphalangeal joint of the index finger was measured when vibrotactile stimuli were applied to the adjacent middle finger and thenar eminence at frequencies and amplitudes that favour activation of rapidly adapting cutaneous afferents. Inputs from Pacinian corpuscles are favoured with high-frequency vibration (300 Hz), while those from Meissner corpuscles are favoured by lower frequencies (30 Hz). Detection of movement was significantly impaired when 300 Hz (20 microm peak-to peak amplitude) complex vibration or 300 Hz (50 microm) sinusoidal vibration was applied to the middle finger and thenar eminence. In contrast, detection of movements was not altered by low-frequency sinusoidal vibration at 30 Hz with an amplitude of 50 microm or with a larger amplitude matched in subjective intensity to the 300 Hz sinusoidal stimulus. Thus it is unlikely that the impairment in detection was due to attention being diverted by vibration of an adjacent digit. In addition, an increase in amplitude of 300 Hz vibration led to a greater impairment of movement detection, so that the impairment was graded with the input. The time taken to nominate the direction of applied movement also increased during 300 Hz but not during 30 Hz sinusoidal vibration. These findings suggest that stimuli which preferentially activate Pacinian, but not Meissner corpuscles, impair proprioceptive acuity in a movement detection task. PMID- 17412775 TI - Respiratory mechanics during exercise in endurance-trained men and women. AB - The purpose of this study was to compare the mechanics of breathing including the measurement of expiratory flow limitation, end-expiratory lung volume, end inspiratory lung volume, and the work of breathing in endurance-trained men (n=8) and women (n=10) during cycle exercise. Expiratory flow limitation was assessed by applying a negative expiratory pressure at the mouth. End-expiratory lung volume and end-inspiratory lung volume were determined by having subjects perform inspiratory capacity manoeuvres. Transpulmonary pressure, taken as the difference between oesophageal and airway opening pressure, was plotted against volume and integrated to determine the work of breathing. Expiratory flow limitation occurred in nine females (90%) and three males (43%) during the final stage of exercise. Females had a higher relative end-expiratory lung volume (42+/-8 versus 35+/-5% forced vital capacity (FVC)) and end-inspiratory lung volume (88+/-5 versus 82+/-7% FVC) compared to males at maximal exercise (P<0.05). Women also had a higher work of breathing compared to men across a range of ventilations. On average, women had a work of breathing that was twice that of men at ventilations above 90 l min(-1). These data suggest that expiratory flow limitation may be more common in females and that they experience greater relative increases in end expiratory lung volume and end-inspiratory lung volume at maximal exercise compared to males. The higher work of breathing in women is probably attributed to their smaller lung volumes and smaller diameter airways. Collectively, these findings suggest that women utilize a greater majority of their ventilatory reserve compared to men and this is associated with a higher cost of breathing. PMID- 17412776 TI - Transpleural ventilation of explanted human lungs. AB - BACKGROUND: The hypothesis that ventilation of emphysematous lungs would be enhanced by communication with the parenchyma through holes in the pleural surface was tested. METHODS: Fresh human lungs were obtained from patients with emphysema undergoing lung transplantation. Control human lungs were obtained from organ donors whose lungs, for technical reasons, were not considered suitable for implantation. Lungs were ventilated through the bronchial tree or transpleurally via a small hole communicating with the underlying parenchyma over which a flanged silicone tube had been cemented to the surface of the lung (spiracle). Measurements included flow-volume-time curves during passive deflation via each pathway; volume of trapped gas recovered from lungs via spiracles when no additional gas was obtainable passively from the airways; and magnetic resonance imaging assessment of spatial distribution of hyperpolarised helium ((3)He) administered through either the airways or spiracles. RESULTS: In emphysematous lungs, passively expelled volumes at 20 s were 94% greater through spiracles than via the airways. Following passive deflation from the airways, an average of 1.07 litres of trapped gas volume was recoverable via spiracles. Regions were ventilated by spiracles that were less well ventilated via bronchi. CONCLUSIONS: Because of the extensive collateral ventilation present in emphysematous lungs, direct communication with the lung parenchyma through non-anatomical pathways has the potential to improve the mechanics of breathing and hence ventilation. PMID- 17412777 TI - Assessment of bronchodilator responsiveness in preschool children using forced oscillations. AB - BACKGROUND: The forced oscillation technique (FOT) requires minimal patient cooperation and is feasible in preschool children. Few data exist on respiratory function changes measured using FOT following inhaled bronchodilators (BD) in healthy young children, limiting the clinical applications of BD testing in this age group. A study was undertaken to determine the most appropriate method of quantifying BD responses using FOT in healthy young children and those with common respiratory conditions including cystic fibrosis, neonatal chronic lung disease and asthma and/or current wheeze. METHODS: A pseudorandom FOT signal (4 48 Hz) was used to examine respiratory resistance and reactance at 6, 8 and 10 Hz; 3-5 acceptable measurements were made before and 15 min after the administration of salbutamol. The post-BD response was expressed in absolute and relative (percentage of baseline) terms. RESULTS: Significant BD responses were seen in all groups. Absolute changes in BD responses were related to baseline lung function within each group. Relative changes in BD responses were less dependent on baseline lung function and were independent of height in healthy children. Those with neonatal chronic lung disease showed a strong baseline dependence in their responses. The BD response in children with cystic fibrosis, asthma or wheeze (based on both group mean data and number of responders) was not greater than in healthy children. CONCLUSIONS: The BD response assessed by the FOT in preschool children should be expressed as a relative change to account for the effect of baseline lung function. The limits for a positive BD response of 40% and 65% for respiratory resistance and reactance, respectively, are recommended. PMID- 17412778 TI - Mechanisms used to restore ventilation after partial upper airway collapse during sleep in humans. AB - BACKGROUND: Most patients with obstructive sleep apnoea (OSA) can restore airflow after an obstructive respiratory event without arousal at least some of the time. The mechanisms that enable this ventilatory recovery are unclear but probably include increased upper airway dilator muscle activity and/or changes in respiratory timing. The aims of this study were to compare the ability to recover ventilation and the mechanisms of compensation following a sudden reduction of continuous positive airway pressure (CPAP) in subjects with and without OSA. METHODS: Ten obese patients with OSA (mean (SD) apnoea-hypopnoea index 62.6 (12.4) events/h) and 15 healthy non-obese non-snorers were instrumented with intramuscular genioglossus electrodes and a mask/pneumotachograph which was connected to a modified CPAP device that could deliver either continuous positive or negative pressure. During stable non-rapid eye movement sleep the CPAP was repeatedly reduced 2-10 cm H2O below the level required to eliminate flow limitation and was held at this level for 5 min or until arousal from sleep occurred. RESULTS: During reduced CPAP the increases in genioglossus activity (311.5 (49.4)% of baseline in subjects with OSA and 315.4 (76.2)% of baseline in non-snorers, p = 0.9) and duty cycle (123.8 (3.9)% of baseline in subjects with OSA and 118.2 (2.8)% of baseline in non-snorers, p = 0.4) were similar in both groups, yet patients with OSA could restore ventilation without cortical arousal less often than non-snorers (54.1% vs 65.7% of pressure drops, p = 0.04). When ventilatory recovery did not occur, genioglossus muscle and respiratory timing changes still occurred but these did not yield adequate pharyngeal patency/ventilation. CONCLUSIONS: Compensatory mechanisms (increased genioglossus muscle activity and/or duty cycle) often restore ventilation during sleep but may be less effective in obese patients with OSA than in non-snorers. PMID- 17412779 TI - Comparison of gel contraction mediated by airway smooth muscle cells from patients with and without asthma. AB - BACKGROUND: Exaggerated bronchial constriction is the most significant and life threatening response of patients with asthma to inhaled stimuli. However, few studies have investigated the contractility of airway smooth muscle (ASM) from these patients. The purpose of this study was to establish a method to measure contraction of ASM cells by embedding them into a collagen gel, and to compare the contraction between subjects with and without asthma. METHODS: Gel contraction to histamine was examined in floating gels containing cultured ASM cells from subjects with and without asthma following overnight incubation while unattached (method 1) or attached (method 2) to casting plates. Smooth muscle myosin light chain kinase protein levels were also examined. RESULTS: Collagen gels containing ASM cells reduced in size when stimulated with histamine in a concentration-dependent manner and reached a maximum at a mean (SE) of 15.7 (1.2) min. This gel contraction was decreased by inhibitors for phospholipase C (U73122), myosin light chain kinase (ML-7) and Rho kinase (Y27632). When comparing the two patient groups, the maximal decreased area of gels containing ASM cells from patients with asthma was 19 (2)% (n = 8) using method 1 and 22 (3)% (n = 6) using method 2, both of which were greater than that of cells from patients without asthma: 13 (2)% (n = 9, p = 0.05) and 10 (4)% (n = 5, p = 0.024), respectively. Smooth muscle myosin light chain kinase levels were not different between the two groups. CONCLUSION: The increased contraction of asthmatic ASM cells may be responsible for exaggerated bronchial constriction in asthma. PMID- 17412780 TI - Protective effect of fruits, vegetables and the Mediterranean diet on asthma and allergies among children in Crete. AB - BACKGROUND: Atopy is not uncommon among children living in rural Crete, but wheeze and rhinitis are rare. A study was undertaken to examine whether this discrepancy could be attributed to a high consumption of fresh fruit and vegetables or adherence to a traditional Mediterranean diet. METHODS: A cross sectional survey was performed in 690 children aged 7-18 years in rural Crete. Parents completed a questionnaire on their child's respiratory and allergic symptoms and a 58-item food frequency questionnaire. Adherence to a Mediterranean diet was measured using a scale with 12 dietary items. Children underwent skin prick tests with 10 common aeroallergens. RESULTS: 80% of children ate fresh fruit (and 68% vegetables) at least twice a day. The intake of grapes, oranges, apples, and fresh tomatoes-the main local products in Crete-had no association with atopy but was protective for wheezing and rhinitis. A high consumption of nuts was found to be inversely associated with wheezing (OR 0.46; 95% CI 0.20 to 0.98), whereas margarine increased the risk of both wheeze (OR 2.19; 95% CI 1.01 to 4.82) and allergic rhinitis (OR 2.10; 95% CI 1.31 to 3.37). A high level of adherence to the Mediterranean diet was protective for allergic rhinitis (OR 0.34; 95% CI 0.18 to 0.64) while a more modest protection was observed for wheezing and atopy. CONCLUSION: The results of this study suggest a beneficial effect of commonly consumed fruits, vegetables and nuts, and of a high adherence to a traditional Mediterranean diet during childhood on symptoms of asthma and rhinitis. Diet may explain the relative lack of allergic symptoms in this population. PMID- 17412781 TI - Urocortin II mediates pro-inflammatory effects in human colonocytes via corticotropin-releasing hormone receptor 2alpha. AB - BACKGROUND/AIMS: Urocortin II (UcnII) is a neuropeptide that binds with high affinity to the corticotropin-releasing hormone receptor 2 (CRHR2) in peripheral tissues. UcnII is synthesised in the intestine, but its role in human intestinal inflammation is largely unknown. METHODS: Responses of human colonic epithelial cells expressing CRHR2 to stimulation by UcnII were measured using ELISA, western blot analysis, real-time reverse transcription-PCR (RT-PCR) and interleukin (IL)8 promoter activity. Expression levels of CRHR2 and UcnII in human colitis were determined by immunofluorescence and real-time RT-PCR in mucosal biopsies from patients with Crohn's and ulcerative colitis, and in human intestinal xenografts after exposure to Clostridium difficile toxin A. RESULTS: It is reported here that expression of CRHR2 mRNA and protein in human colonic epithelial cells (HT 29) are increased by exposure to C difficile toxin A or tumour necrosis factor (TNF)alpha. Stimulation of non-transformed NCM460 colonocytes overexpressing CRHR2alpha receptor with UcnII resulted in a time- and concentration-dependent increase in IL8 production. UcnII stimulation also led to activation of nuclear factor-kappaB (NF-kappaB) and mitogen-acivated protein (MAP) kinase in these cells, as evidenced by degradation of IkappaBalpha and phosphorylation of the p65 subunit of NF-kappaB and extracellularly regulated kinase (ERK) 1/2. Furthermore, expression of UcnII and CRHR2 mRNA was increased in mucosal samples of patients with inflammatory bowel disease, and after exposure of human intestinal xenografts to C difficile toxin A. CONCLUSIONS: These results suggest that UcnII has pro-inflammatory effects in human intestinal cells via the CRHR2alpha receptor and may play an important role in the pathophysiology of colitis in humans. PMID- 17412782 TI - Refinements in modeling the passive properties of laryngeal soft tissue. AB - The nonlinear viscoelastic passive properties of three canine intrinsic laryngeal muscles, the lateral cricoarytenoid (LCA), the posterior cricoarytenoid (PCA), and the interarytenoid (IA), were fit to the parameters of a modified Kelvin model. These properties were compared with those of the thyroarytenoid (TA) and cricothyroid (CT) muscles, as well as previously unpublished viscoelastic characteristics of the human vocal ligament. Passive parameters of the modified Kelvin model were summarized for the vocal ligament, mucosa, and all five laryngeal muscles. Results suggest that the LCA, PCA, and IA muscles are functionally different from the TA and CT muscles in their load-bearing capacity. Furthermore, the LCA, PCA, and IA have a much larger stress-strain hysteresis effect than has been previously reported for the TA and CT or the vocal ligament. The variation in this effect suggests that the connective tissue within the TA and CT muscles is somehow similar to the vocal ligament but different from the LCA, PCA, or IA muscles. Further demonstrating the potential significance of grouping tissues in the laryngeal system by functional groups in the laryngeal system was the unique finding that, over their working elongation range, the LCA and PCA were nearly as exponentially stiff as the vocal ligament. This paper was written in conjunction with an online technical report (http://www.ncvs.org/ncvs/library/tech) in which comprehensive muscle data and sensitivity analysis, as well as downloadable data files and computer scripts, are made available. PMID- 17412783 TI - Utility of circulating IGF-I as a biomarker for assessing body composition changes in men during periods of high physical activity superimposed upon energy and sleep restriction. AB - Insulin-like growth factor (IGF)-I is a biomarker that may have greater utility than other conventional nutritional biomarkers in assessing nutritional, health, and fitness status. We hypothesized that the IGF-I system would directionally track a short-term energy deficit and would be more related to changes in body composition than other nutritional biomarkers. Thirty-five healthy men (24 +/- 0.3 yr) underwent 8 days of exercise and energy imbalance. Total and free IGF-I, IGF binding proteins-1, -2, and -3, the acid labile subunit, transferrin, ferritin, retinol binding protein, prealbumin, testosterone, triiodothyronine, thyroxine, and leptin responses were measured. Dual-energy X-ray absorptiometry assessed changes in body mass and composition. Repeated-measures ANOVA, correlation analysis, and receiver operator characteristic curves were used for statistical analyses (P < or = 0.05). Body mass (-3.8%), fat-free mass (-2.2%), and fat mass (-12.9%) all decreased. Total and free IGF-I, IGF binding protein-3, and the acid labile subunit and prealbumin, but not transferrin, retinol-binding protein, and ferritin, directionally tracked the energy deficit and losses in body composition. The correlation (r = 0.43) between changes in free IGF-I and body and fat-free mass was the only significant association observed. Receiver operator characteristic curve analysis revealed that a baseline value < 1.67 for the molar volume ratio of IGF-I to acid labile subunit had an area under the curve of 0.745 and was a significant discriminator for those subjects losing >5% body mass. The IGF-I system is an important adjunct in the overall assessment of adaptation to stress imposed by high levels of physical activity superimposed on energy and sleep restriction and is more closely associated with losses in body mass and fat-free mass than other conventional nutritional biomarkers. PMID- 17412784 TI - The effects of hypoxia on load compensation during sustained incremental resistive loading in patients with obstructive sleep apnea. AB - Inspiratory load compensation is impaired in patients with obstructive sleep apnea (OSA), a condition characterized by hypoxia during sleep. We sought to compare the effects of sustained hypoxia on ventilation during inspiratory resistive loading in OSA patients and matched controls. Ten OSA patients and 10 controls received 30 min of isocapnic hypoxia (arterial oxygen saturation 80%) and normoxia in random order. Following the gas period, subjects were administered six incremental 2-min inspiratory resistive loads while breathing room air. Ventilation was measured throughout the loading period. In both patients and controls, there was a significant increase in inspiratory time with increasing load (P = 0.006 and 0.003, respectively), accompanied by a significant fall in peak inspiratory flow (P = 0.006 and P < 0.001, respectively). The result was a significant fall in minute ventilation in both groups with increasing load (P = 0.003 and P < 0.001, respectively). There was no difference between the two groups for these parameters. The only difference between the two groups was a transient increase in tidal volume in controls (P = 0.02) but not in OSA patients (P = 0.57) during loading. Following hypoxia, there was a significant increase in minute ventilation during loading in both groups (P < 0.001). These results suggest that ventilation during incremental resistive loading is preserved in OSA patients and that it appears relatively impervious to the effects of hypoxia. PMID- 17412785 TI - Decreased left ventricular function, myocarditis, and coronary arteriolar medial thickening following monocrotaline administration in adult rats. AB - Decreased right as well as left ventricular function can be associated with pulmonary hypertension (PH). Numerous investigations have examined cardiac function following induction of pulmonary hypertension with monocrotaline (MCT) assuming that MCT has no direct cardiac effect. We tested this assumption by examining left ventricular function and histology of isolated and perfused hearts from MCT-treated rats. Experiments were performed on 50 male Sprague-Dawley rats [348 +/- 6 g (SD)]. Thirty-seven rats received MCT (50 mg/kg sc; MCT group) while the remainder did not (Control group). Three weeks later, pulmonary artery pressure was assessed echocardiographically in 20 MCT and 8 Control rats. The hearts were then excised and perfused in the constant pressure Langendorff mode to determine peak left ventricular pressure (LVP), the peak instantaneous rate of pressure increase (+dP/dtmax) and decrease (-dP/dtmax), as well as the rate pressure product (RPP). Histological sections were subsequently examined. Pulmonary artery pressure was higher in the MCT-treated group compared with the Control group [12.9 +/- 6 vs. 51 +/- 35.3 mmHg (P < 0.01)]. Left ventricular systolic function and diastolic relaxation were decreased in the MCT group compared with the Control group (+dP/dtmax 4,178 +/- 388 vs. 2,801 +/- 503 mmHg/s, LVP 115 +/- 11 vs. 83 +/- 14 mmHg, RPP 33,688 +/- 1,910 vs. 23,541 +/- 3,858 beats x min(-1) x mmHg(-1), -dP/dtmax -3,036 +/- 247 vs. -2,091 +/- 389 mmHg/s; P < 0.0001). The impairment of cardiac function was associated with myocarditis and coronary arteriolar medial thickening. Similarly depressed ventricular function and inflammatory infiltration was seen in 12 rats 7 days after MCT administration. Our findings appear unrelated to the degree of PH and indicate a direct cardiotoxic effect of MCT. PMID- 17412786 TI - WISE 2005: stroke volume changes contribute to the pressor response during ischemic handgrip exercise in women. AB - The mechanism of the pressor response to small muscle mass (e.g., forearm) exercise and during metaboreflex activation may include elevations in cardiac output (Q) or total peripheral resistance (TPR). Increases in Q must be supported by reductions in visceral venous volume to sustain venous return as heart rate (HR) increases. Therefore, this study tested the hypothesis that increases in Q, supported by reductions in splanchnic volume (portal vein constriction), explain the pressor response during handgrip exercise and metaboreflex activation. Seventeen healthy women performed 2 min of static ischemic handgrip exercise and 2 min of postexercise circulatory occlusion (PECO) while HR, stroke volume and superficial femoral artery flow (Doppler), blood pressure (Finometer), portal vein diameter (ultrasound imaging), and muscle sympathetic nerve activity (MSNA; microneurography) were measured followed by the calculation of Q, TPR, and leg vascular resistance (LVR). Compared with baseline, mean arterial blood pressure (MAP) (P < 0.001) and Q (P < 0.001) both increased in each minute of exercise accompanied by a approximately 5% reduction in portal vein diameter (P < 0.05). MAP remained elevated during PECO, whereas Q decreased below exercise levels. MSNA was elevated above baseline during the second minute of exercise and through the PECO period (P < 0.05). Neither TPR nor LVR was changed from baseline during exercise and PECO. The data indicate that the majority of the blood pressure response to isometric handgrip exercise in women was due to mobilization of central blood volume and elevated stroke volume and Q rather than elevations in TVR or LVR resistance. PMID- 17412787 TI - Unloaded rat Achilles tendons continue to grow, but lose viscoelasticity. AB - Tendons can function as springs and thereby preserve energy during cyclic loading. They might also have damping properties, which, hypothetically, could reduce risk of microinjuries due to fatigue at sites of local stress concentration within the tendon. At mechanical testing, damping will appear as hysteresis. How is damping influenced by training or disuse? Does training decrease hysteresis, thereby making the tendon a better spring, or increase hysteresis and thus improve damping? Seventy-eight female 10-wk-old Sprague Dawley rats were randomized to three groups. Two groups had botulinum toxin injected into the calf muscles to unload the left Achilles tendon through muscle paralysis. One of these groups was given doxycycline, as a systemic matrix metalloproteinase inhibitor. The third group served as loaded controls. The Achilles tendons were harvested after 1 or 6 wk for biomechanical testing. An increase with time was seen in tendon dry weight, wet weight, water content, transverse area, length, stiffness, force at failure, and energy uptake in all three groups (P < 0.001 for each parameter). Disuse had no effect on these parameters. Creep was decreased with time in all groups. The only significant effect of disuse was on hysteresis (P = 0.004) and creep (P = 0.007), which both decreased with disuse compared with control, and on modulus, which was increased (P = 0.008). Normalized glycosaminoglycan content was unaffected by time and disuse. No effect of doxycycline was observed. The results suggest that in growing animals, the tendons continue to grow regardless of mechanical loading history, whereas maintenance of damping properties requires mechanical stimulation. PMID- 17412788 TI - Quantitative trait loci associated with maximal exercise endurance in mice. AB - The role of genetics in the determination of maximal exercise endurance is unclear. Six- to nine-week-old F2 mice (n = 99; 60 female, 39 male), derived from an intercross of two inbred strains that had previously been phenotyped as having high maximal exercise endurance (Balb/cJ) and low maximal exercise endurance (DBA/2J), were treadmill tested to estimate exercise endurance. Selective genotyping of the F2 cohort (n = 12 high exercise endurance; n = 12 low exercise endurance) identified a significant quantitative trait locus (QTL) on chromosome X (53.7 cM, DXMit121) in the entire cohort and a suggestive QTL on chromosome 8 (36.1 cM, D8Mit359) in the female mice. Fine mapping with the entire F2 cohort and additional informative markers confirmed and narrowed the QTLs. The chromosome 8 QTL (EE8(F)) is homologous with two suggestive human QTLs and one significant rat QTL previously linked with exercise endurance. No effect of sex (P = 0.33) or body weight (P = 0.79) on exercise endurance was found in the F2 cohort. These data indicate that genetic factors in distinct chromosomal regions may affect maximal exercise endurance in the inbred mouse. Whereas multiple genes are located in the identified QTL that could functionally affect exercise endurance, this study serves as a foundation for further investigations delineating the identity of genetic factors influencing maximum exercise endurance. PMID- 17412789 TI - Stimulation pulse characteristics and electrode configuration determine site of excitation in isolated mammalian skeletal muscle: implications for fatigue. AB - We examined whether electrical field stimulation with varying characteristics could excite isolated mammalian skeletal muscle through different sites. Supramaximal (20-V, 0.1-ms) pulse stimulation with transverse wire or parallel plate electrodes evoked similar forces in nonfatigued slow-twitch soleus and fast twitch extensor digitorum longus (EDL) muscles from mice. d-tubocurarine shifted the twitch force-stimulation strength relationship toward higher pulse strengths with both electrode configurations in soleus muscle, suggesting that weaker pulses excite muscle via neuromuscular transmission. With wire stimulation, movement of the recording electrode along the muscle caused a delay between the stimulus artifact and the peak of the action potential, consistent with action potential propagation along the sarcolemma. TTX abolished all contractions evoked with 20-V, 0.1-ms pulses, suggesting that excitation occurred via voltage dependent Na+ channels and, hence, muscle action potentials. TTX did not prevent force development with > or = 0.4-ms pulses in soleus or 1-ms pulses in EDL muscle. Furthermore, myoplasmic Ca2+ (i.e., the fura 2 ratio) and sarcomere shortening were greater during tetanic stimulation with 2.0-ms than with 0.5-ms pulses in flexor digitorum brevis fibers from rats. TTX prevented all shortening and Ca2+ release with 0.5-ms, but not 2.0-ms, pulses, indicating that longer pulses can directly trigger Ca2+ release. Hence, proper interpretation of mechanistic studies requires precise understanding of how muscles are excited; otherwise, incorrect conclusions can be made. Using this new understanding, we showed that disrupted propagation of action potentials along the surface membrane is a major cause of fatigue in soleus muscle that is focally and continuously stimulated at 125 Hz. PMID- 17412790 TI - Protection of muscle membrane excitability during prolonged cycle exercise with glucose supplementation. AB - To determine if exercise-induced depressions in neuromuscular function are altered with oral glucose supplementation, 15 untrained participants (Vo2 peak = 45 +/- 2 ml x kg(-1) x min(-1), mean +/- SE) performed prolonged cycle exercise at approximately 60% Vo2 peak on two occasions: without glucose supplementation (NG) and with oral glucose supplementation (G). The oral G began at 30 min of exercise and was administered every 15 min (total ingested = 1.23 +/- 0.11 g carbohydrate/kg body mass). Quadriceps isometric properties and membrane excitability were assessed prior to exercise, after 90 min of exercise, and at fatigue. Cycle time to fatigue was greater (P < 0.05) in G compared with NG (137 +/- 7 vs. 115 +/- 6 min). Progressive reductions (P < 0.05) in maximal voluntary contraction (MVC, N) were observed for NG at 90 min (441 +/- 29) and at fatigue (344 +/- 33) compared with pre-exercise (666 +/- 30). At fatigue in G, the reduction in MVC was not as pronounced (P < 0.05) as in NG. Motor unit activation assessed with the interpolated twitch technique during an MVC following exercise was not different between conditions. During cycling, the G condition also resulted in a higher (P < 0.05) muscle compound potential (M-wave) amplitude (mV) at both 90 min (+50%) and at fatigue (+87%) compared with NG. Similar effects were also found M-wave area (mV/ms). These results suggest that the ergogenic effect of glucose supplementation occurs not as a result of decreased neural activation but to improved muscle function, possibly as a consequence of protection of muscle membrane excitability. PMID- 17412791 TI - Experimental quadriceps muscle pain impairs knee joint control during walking. AB - Pain is a cardinal symptom in musculoskeletal diseases involving the knee joint, and aberrant movement patterns and motor control strategies are often present in these patients. However, the underlying neuromuscular mechanisms linking pain to movement and motor control are unclear. To investigate the functional significance of muscle pain on knee joint control during walking, three dimensional gait analyses were performed before, during, and after experimentally induced muscle pain by means of intramuscular injections of hypertonic saline (5.8%) into vastus medialis (VM) muscle of 20 healthy subjects. Isotonic saline (0.9%) was used as control. Surface electromyography (EMG) recordings of VM, vastus lateralis (VL), biceps femoris, and semitendinosus muscles were synchronized with the gait analyses. During experimental muscle pain, the loading response phase peak knee extensor moments were attenuated, and EMG activity in the VM and VL muscles was reduced. Compressive forces, adduction moments, knee joint kinematics, and hamstring EMG activity were unaffected by pain. Interestingly, the observed changes persisted when the pain had vanished. The results demonstrate that muscle pain modulated the function of the quadriceps muscle, resulting in impaired knee joint control and joint instability during walking. The changes are similar to those observed in patients with knee pain. The loss of joint control during and after pain may leave the knee joint prone to injury and potentially participate in the chronicity of musculoskeletal problems, and it may have clinically important implications for rehabilitation and training of patients with knee pain of musculoskeletal origin. PMID- 17412792 TI - Human lung density is not altered following normoxic and hypoxic moderate intensity exercise: implications for transient edema. AB - The purpose of this study was to examine the effects of exercise on extravascular lung water as it may relate to pulmonary gas exchange. Ten male humans underwent measures of maximal oxygen uptake (Vo2 max) in two conditions: normoxia (N) and normobaric hypoxia of 15% O2 (H). Lung density was measured by quantified MRI before and 48.0 +/- 7.4 and 100.7 +/- 15.1 min following 60 min of cycling exercise in N (intensity = 61.6 +/- 9.5% Vo2 max) and 55.5 +/- 9.8 and 104.3 +/- 9.1 min following 60 min cycling exercise in H (intensity = 65.4 +/- 7.1% hypoxic Vo2 max), where Vo2 max = 65.0 +/- 7.5 ml x kg(-1) x min(-1) (N) and 54.1 +/- 7.0 ml x kg(-1) x min(-1) (H). Two subjects demonstrated mild exercise-induced arterial hypoxemia (EIAH) [minimum arterial oxygen saturation (SaO2 min) = 94.5% and 93.8%], and seven subjects demonstrated moderate EIAH (SaO2 min = 91.4 +/- 1.1%) as measured noninvasively during the Vo2 max test in N. Mean lung densities, measured once preexercise and twice postexercise, were 0.177 +/- 0.019, 0.181 +/- 0.019, and 0.173 +/- 0.019 g/ml (N) and 0.178 +/- 0.021, 0.174 +/- 0.022, and 0.176 +/- 0.019 g/ml (H), respectively. No significant differences (P > 0.05) were found in lung density following exercise in either condition or between conditions. Transient interstitial pulmonary edema did not occur following sustained steady-state cycling exercise in N or H, indicating that transient edema does not result from pulmonary capillary leakage during sustained submaximal exercise. PMID- 17412793 TI - The biphasic force-velocity relationship in whole rat skeletal muscle in situ. AB - Edman has reported that the force-velocity relationship (FVR) departs from Hill's classic hyperbola near 0.80 of measured isometric force (J Physiol 404: 301-321, 1988). The purpose of this study was to investigate the biphasic nature of the FVR in the rested state and after some recovery from fatigue in the rat medial gastrocnemius muscle in situ. Force-velocity characteristics were determined before and during recovery from fatigue induced by intermittent stimulation at 170 Hz for 100 ms each second for 6 min. Force-velocity data were obtained for isotonic contractions with 100 ms of 200-Hz stimulation, including several measurements with loads above 0.80 of measured isometric force. The force velocity data obtained in this study were fit well by a double-hyperbolic equation. A departure from Hill's classic hyperbola was found at 0.88+/-0.01 of measured isometric force, which is higher than the approximately 0.80 reported by Edman et al. for isolated frog fibers. After 45 min of recovery, maximum shortening velocity was 86+/-2% of prefatigue, but neither curvature nor predicted isometric force was significantly different from prefatigue. The location of the departure from Hill's classic hyperbola was not different after this recovery from the fatiguing contractions. Including an isometric point in the data set will not yield the same values for maximal velocity and the degree of curvature as would be obtained using the double hyperbola approach. Data up to 0.88 of measured isometric force can be used to fit data to the Hill equation. PMID- 17412794 TI - Insulin-like growth factor I stimulates recovery of bone lost after a period of skeletal unloading. AB - IGF-I stimulates osteoblast proliferation, bone formation, and increases bone volume in normal weight-bearing animals. During skeletal unloading or loss of weight bearing, bone becomes unresponsive to the anabolic effects of insulin-like growth factor I (IGF-I). To determine whether skeletal reloading after a period of unloading increases bone responsiveness to IGF-I, we examined bone structure and formation in response to IGF-I under different loading conditions. Twelve week-old rats were divided into six groups: loaded (4 wk), unloaded (4 wk), and unloaded/reloaded (2/2 wk), and treated with IGF-I (2.5 mg x kg(-1) x day(-1)) or vehicle during the final 2 wk. Cortical bone formation rate (BFR), cancellous bone volume and architecture in the secondary spongiosa (tibia and vertebrae), and total volume and calcified volume in the primary spongiosa (tibia) were assessed. Periosteal BFR decreased during unloading, remained low during reloading in the vehicle-treated group, but was dramatically increased in IGF-I treated animals. Cancellous bone volume decreased with unloading and increased with reloading, but the effect was exaggerated in the tibia of IGF-I-treated animals. Total and calcified volumes in the primary spongiosa decreased during unloading in the vehicle-treated animals. IGF-I treatment prevented the loss in volume. These data show that reloading after a period of skeletal unloading increases bone responsiveness to IGF-I, and they suggest that IGF-I may be of therapeutic use in patients who have lost bone as a consequence of prolonged skeletal disuse. PMID- 17412795 TI - Persistence of eupnea and gasping following blockade of both serotonin type 1 and 2 receptors in the in situ juvenile rat preparation. AB - In severe hypoxia or ischemia, normal eupneic breathing is replaced by gasping, which can serve as a powerful mechanism for "autoresuscitation." We have proposed that gasping is generated by medullary neurons having intrinsic pacemaker bursting properties dependent on a persistent sodium current. A number of neuromodulators, including serotonin, influence persistent sodium currents. Thus we hypothesized that endogenous serotonin is essential for gasping to be generated. To assess such a critical role for serotonin, a preparation of the perfused, juvenile in situ rat was used. Activities of the phrenic, hypoglossal, and vagal nerves were recorded. We added blockers of type 1 and/or type 2 classes of serotonergic receptors to the perfusate delivered to the preparation. Eupnea continued following additions of any of the blockers. Changes were limited to an increase in the frequency of phrenic bursts and a decline in peak heights of all neural activities. In ischemia, gasping was induced following any of the blockers. Few statistically significant changes in parameters of gasping were found. We thus did not find a differential suppression of gasping, compared with eupnea, following blockers of serotonin receptors. Such a differential suppression had been proposed based on findings using an in vitro preparation. We hypothesize that multiple neurotransmitters/neuromodulators influence medullary mechanisms underlying the neurogenesis of gasping. In greatly reduced in vitro preparations, the importance of any individual neuromodulator, such as serotonin, may be exaggerated compared with its role in more intact preparations. PMID- 17412796 TI - Preserved reflex cutaneous vasodilation in cystic fibrosis does not include an enhanced nitric oxide-dependent mechanism. AB - In humans, vasoactive intestinal peptide (VIP) may play a role in reflex cutaneous vasodilation during body heating. We tested the hypothesis that the nitric oxide (NO)-dependent contribution to active vasodilation is enhanced in the skin of subjects with cystic fibrosis (CF), compensating for sparse levels of VIP. In 2 parallel protocols, microdialysis fibers were placed in the skin of 11 subjects with CF and 12 controls. Lactated Ringer was perfused at one microdialysis site and NG-nitro-L-arginine methyl ester (2.7 mg/ml) was perfused at a second microdialysis site. Skin blood flow was monitored over each site with laser-Doppler flowmetry. In protocol 1, local skin temperature was increased 0.5 degrees C every 5 s to 42 degrees C, and then it maintained at 42 degrees C for approximately 45 min. In protocol 2, subjects wore a tube-lined suit perfused with water at 50 degrees C, sufficient to increase oral temperature (Tor) 0.8 degrees C. Cutaneous vascular conductance (CVC) was calculated (flux/mean arterial pressure) and scaled as percent maximal CVC (sodium nitroprusside; 8.3 mg/ml). Vasodilation to local heating was similar between groups. The change (Delta%CVCmax) in CVC with NO synthase inhibition on the peak (9+/-3 vs. 12+/ 5%CVCmax; P=0.6) and the plateau (45+/-3 vs. 35+/-5%CVCmax; P=0.1) phase of the skin blood flow response to local heating was similar in CF subjects and controls, respectively. Reflex cutaneous vasodilation increased CVC in CF subjects (58+/-4%CVCmax) and controls (53+/-4%CVCmax; P=0.37) and NO synthase inhibition attenuated CVC in subjects with CF (37+/-6%CVCmax) and controls (35+/ 5%CVCmax; P=0.8) to a similar degree. Thus the preservation of cutaneous active vasodilation in subjects with CF is not associated with an enhanced NO-dependent vasodilation. PMID- 17412797 TI - Development and multicenter evaluation of the N latex CDT direct immunonephelometric assay for serum carbohydrate-deficient transferrin. AB - BACKGROUND: Carbohydrate-deficient transferrin (CDT) is a promising biomarker of alcohol abuse. We describe the development and multicenter evaluation of N Latex CDT (Dade Behring), an automated, particle-enhanced, homogeneous immunonephelometric assay for directly determining CDT. METHODS: N Latex CDT uses a monoclonal antibody that recognizes the structure of transferrin glycoforms lacking 1 or 2 complete N-glycans [i.e., disialo-, monosialo-, and asialotransferrins (CDT glycoforms)] in combination with a simultaneous assay for total transferrin. The Dade Behring BN II and BN ProSpec systems automatically calculate the CDT value as a percentage of total transferrin (%CDT). No preanalytical sample treatment is used. RESULTS: Total imprecision values for serum pools containing 1.8%-8.7% CDT were 3.4%-10.4% (mean, 6.8%). The mean (SD) %CDT for 561 serum samples from healthy control individuals was 1.76% (0.27%; range, 1.01%-2.85%). No marked sex or age differences were noted. The 97.5th percentile was at 2.35%. Transferrin genetic variants did not interfere with measurements. High transferrin concentrations did not falsely increase %CDT values, but increased %CDT values were noted for some samples with transferrin concentrations <1.1 g/L. N Latex CDT results correlated with those of a commercial CDT immunoassay involving column separation (r(2) = 0.862) and an HPLC candidate reference method (r(2) = 0.978). CONCLUSION: N Latex CDT is the first direct immunoassay for quantifying %CDT in serum. The specificity of N Latex CDT for identifying alcohol abuse may be higher than for immunoassays that use column separation, because transferrin genetic variants do not interfere with measurements. PMID- 17412798 TI - Evaluation of a new pooling strategy based on leukocyte count for rapid quantification of allele frequencies. AB - BACKGROUND: Allele frequencies of single-nucleotide polymorphisms (SNPs) can be quantified from DNA pools. The conventional preparation of DNA pools requires DNA isolation and quantification for each blood sample. We hypothesized that pooling of whole blood samples according to their leukocyte count, which determines DNA content, would be as reliable as the conventional pooling method but much less tedious to perform. METHODS: We collected 100 whole blood samples and measured the leukocyte count. Samples were frozen until further use. After thawing, pools were generated by combining aliquots containing an equal number of leukocytes. In parallel, DNA was extracted from another aliquot, DNA concentration was measured, and DNA concentration-based pools were assembled. All original samples were genotyped directly using 4 different SNP assays to obtain the exact allele frequencies in the pool. In addition, samples of known genotypes were mixed according to the DNA concentration or the leukocyte count to generate artificial samples of known allele frequencies. We analyzed pools and mixes in triplicate by pyrosequencing and calculated allelic frequencies. RESULTS: Leukocyte and DNA pooling provided equally accurate and precise SNP frequencies comparable to published data. CONCLUSION: DNA and leukocyte pooling are both suitable strategies to determine allele frequencies in frozen samples. The leukocyte pooling approach is much less tedious, quicker, and less expensive. It should be always considered if leukocyte counts are available. PMID- 17412799 TI - Genetic and environmental influences on plasma homocysteine: results from a Danish twin study. AB - BACKGROUND: Increased plasma homocysteine has been linked to many clinical conditions including atherosclerosis and ischemic stroke. We assessed the genetic and environmental influences on homocysteine in adult twins and tested the influence of 3 candidate polymorphisms. METHODS: Homocysteine was analyzed in 1206 healthy twins, who were genotyped for 3 polymorphisms: MTHFR 677C>T, MTR 2756A>G, and NNMT (dbSNP: rs694539). To perform quantitative trait linkage analysis of the MTHFR locus, the genotyping was supplemented with 2 genetic markers localized on each site of the MTHFR locus. The twin data were analyzed using biometric structural equation models as well as a combined association and linkage analysis in 2 age cohorts. RESULTS: Age, sex, and MTHFR genotype have a significant impact on homocysteine concentrations, whereas the other genotypes were not associated with homocysteine concentrations. The variance in homocysteine could be solely ascribed to additive genetic and nonshared environmental factors, with an estimated additive genetic proportion of total variation at age 18-39 years of 0.63 (95% CI, 0.53-0.71) and at age 40-65 years of 0.27 (95% CI, 0.10-0.41). The impact of the MTHFR locus is estimated to explain 53% (95% CI, 0.07-0.67) of the total phenotypic variation in persons 18 39 years old and 24% (95% CI, 0.00-0.39) in persons 40-65 years old, i.e., almost all additive genetic variance. CONCLUSIONS: Homocysteine concentrations have a high heritability that decreases with age. The MTHFR gene locus is responsible for almost all the variation attributable to genetic factors, leaving very little influence of other genetic variations. PMID- 17412800 TI - Effect of continuous intravenous administration of human metastin 45-54 on the neuroendocrine activity of the hypothalamic-pituitary-testicular axis in the adult male rhesus monkey (Macaca mulatta). AB - In agonadal juvenile male monkeys, continuous administration of human metastin 45 54 (hu metastin 45-54) leads to desensitization of its receptor, G protein coupled receptor 54 (GPR54), and decreased LH. The present study extended this observation to the adult male monkey, a more preclinically relevant model in which robust activity in the hypothalamic-pituitary-testicular axis is present. Continuous iv infusion of hu metastin 45-54 at either 200 or 400 microg/h elicited a marked rise in circulating LH that peaked 2-3 h after initiation of treatment. Thereafter, levels declined, and by 24 h, LH in metastin 45-54-infused animals was similar to control. LH release in response to an iv bolus of hu metastin 45-54 (10-30 microg) during the final 3 h of continuous infusion was truncated or abolished (low and high peptide dose, respectively). GPR54 desensitization by the high-dose metastin 45-54 infusion was associated with compromised pituitary response to a bolus GnRH injection (0.3 microg). LH pulse amplitude and pulse frequency were markedly suppressed during high-dose metastin 45-54 treatment. Surprisingly, the fidelity of the relationship between circulating testosterone (T) and LH was distorted during the high-dose peptide infusion. Thus, for a given concentration of LH, T levels were invariably higher during the high-dose metastin 45-54 infusion than during vehicle, suggesting that the peptide may exert direct actions on the testis to amplify T production. These findings support the notion that GPR54 is desensitized by continuous exposure to ligand, and they raise the possibility of an intratesticular role of GPR54. PMID- 17412801 TI - Direct regulation of androgen receptor-associated protein 70 by thyroid hormone and its receptors. AB - Thyroid hormone (T3) regulates multiple physiological processes during development, growth, differentiation, and metabolism. Most T3 actions are mediated via thyroid hormone receptors (TRs) that are members of the nuclear hormone receptor superfamily of ligand-dependent transcription factors. The effects of T3 treatment on target gene regulation was previously examined in TRalpha1-overexpressing hepatoma cell lines (HepG2-TRalpha1). Androgen receptor (AR)-associated protein 70 (ARA70) was one gene found to be up-regulated by T3. The ARA70 is a ligand-dependent coactivator for the AR and was significantly increased by 4- to 5-fold after T3 treatment by Northern blot analyses in the HepG2-TRalpha1 stable cell line. T3 induced a 1- to 2-fold increase in the HepG2 TRbeta1 stable cell line. Both stable cell lines attained the highest fold expression after 24 h treatment with 10 nM T3. The ARA70 protein was increased up to 1.9-fold after T3 treatment in HepG2-TRalpha1 cells. Similar findings were obtained in thyroidectomized rats after T3 application. Cycloheximide treatment did not suppress induction of ARA70 transcription by T3, suggesting that this regulation is direct. A series of deletion mutants of ARA70 promoter fragments in pGL2 plasmid were generated to localize the thyroid hormone response element (TRE). The DNA fragments (-234/-190 or +56/+119) gave 1.55- or 2-fold enhanced promoter activity by T3. Thus, two TRE sites exist in the upstream-regulatory region of ARA70. The TR-TRE interaction was further confirmed with EMSAs. Additionally, ARA70 could interfere with TR/TRE complex formation. Therefore, the data indicated that ARA70 suppresses T3 signaling in a TRE-dependent manner. These experimental results suggest that T3 directly up-regulates ARA70 gene expression. Subsequently, ARA70 negatively regulates T3 signaling. PMID- 17412802 TI - Focal adhesion kinase and paxillin: novel regulators of brain sexual differentiation? AB - Steroid-mediated sexual differentiation of the brain is a developmental process that permanently organizes the brain into a male or female phenotype. Previous studies in the rodent have examined the steroid-mediated mechanisms of male brain development. In an effort to identify molecules involved in female brain development, a high-throughput proteomics approach called PowerBlot was used to identify signaling proteins differentially regulated in the neonatal male and female rat hypothalamus during the critical period for brain sexual differentiation. Focal adhesion kinase (FAK) and paxillin, both members of the focal adhesion complex family of proteins, were significantly elevated in the newborn female compared with the male hypothalamus. Sex differences in these proteins were not detected in brain regions that are not subject to substantial organizational effects of steroids. Estrogens, the aromatized products of testosterone in the male, can both masculinize and defeminize the male brain. Daily estradiol administration to neonatal females significantly reduced FAK and paxillin in the hypothalamus, and aromatase inhibition increased paxillin in males to levels comparable with females. Androgens also appear to modulate paxillin levels in combination with estrogen action. Across development, hypothalamic levels of FAK were significantly elevated in females compared with males on postnatal d 6. Synaptic circuits in the hypothalamus develop sex differences perinatally. Estradiol treatment of cultured hypothalamic neurons significantly enhanced axon branching (P<0.01), consistent with the phenotype of FAK-deficient neurons. Together, these data implicate FAK and paxillin as regulators of sex differences in neuronal morphology. PMID- 17412803 TI - Melanocortinergic activation by melanotan II inhibits feeding and increases uncoupling protein 1 messenger ribonucleic acid in the developing rat. AB - The hypothalamic neurocircuitry that regulates energy homeostasis in adult rats is not fully developed until the third postnatal week. In particular, fibers from the hypothalamic arcuate nucleus, including both neuropeptide Y (NPY) and alpha MSH fibers, do not begin to innervate downstream hypothalamic targets until the second postnatal week. However, alpha-MSH fibers from the brainstem and melanocortin receptors are present in the hypothalamus at birth. The present study investigated the melanocortin system in the early postnatal period by examining effects of the melanocortin receptor agonist melanotan II (MTII) on body weight, energy expenditure, and hypothalamic NPY expression. Rat pups were injected ip with MTII (3 mg/kg body weight) or saline on postnatal day (P) 5 to P6, P10-P11, or P15-P16 at 1700 and 0900 h and then killed at 1300 h. Stomach weight and brown adipose tissue uncoupling protein 1 mRNA were determined. In addition, we assessed central c-Fos activation 90 min after MTII administration and hypothalamic NPY mRNA after twice daily MTII administration from P5-P10 or P10-P15. MTII induced hypothalamic c-Fos activation as well as attenuating body weight gain in rat pups. Stomach weight was significantly decreased and uncoupling protein 1 mRNA was increased at all ages, indicating decreased food intake and increased energy expenditure, respectively. However, MTII had no effect on NPY mRNA levels in any hypothalamic region. These findings demonstrate that MTII can inhibit food intake and stimulate energy expenditure before the full development of hypothalamic feeding neurocircuitry. These effects do not appear to be mediated by changes in NPY expression. PMID- 17412804 TI - Dehydration-induced proteome changes in the rat hypothalamo-neurohypophyseal system. AB - The hypothalamo-neurohypophyseal system (HNS) mediates neuroendocrine responses to dehydration through the action of the antidiuretic hormone vasopressin (VP). VP is synthesized as part of a prepropeptide in magnocellular neurons of the hypothalamic supraoptic nucleus (SON) and paraventricular nucleus. This precursor is processed during transport to axon terminals in the posterior pituitary gland, in which biologically active VP is stored until mobilized for secretion by electrical activity evoked by osmotic cues. During release, VP travels through the blood stream to specific receptor targets located in the kidney in which it increases the permeability of the collecting ducts to water, reducing the renal excretion of water, thus promoting water conservation. The HNS undergoes a dramatic function-related plasticity during dehydration. We hypothesize that alterations in steady-state protein levels might be partially responsible for this remodeling. We investigated dehydration-induced changes in the SON and pituitary neurointermediate lobe (NIL) proteomes using two-dimensional fluorescence difference gel electrophoresis. Seventy proteins were altered by dehydration, including 45 in the NIL and 25 in the SON. Using matrix-assisted laser desorption/ionization mass spectrometry, we identified six proteins in the NIL (four down, two up) and nine proteins in the SON (four up, five down) that are regulated as a consequence of chronic dehydration. Results for five of these proteins, namely Hsp1alpha (heat shock protein 1alpha), NAP22 (neuronal axonal membrane protein 22), GRP58 (58 kDa glucose regulated protein), calretinin, and ProSAAS (proprotein convertase subtilisin/kexin type 1 inhibitor), have been confirmed using independent methods such as semiquantitative Western blotting, two-dimensional Western blotting, enzyme-linked immunoassay, and immunohistochemistry. These proteins may have roles in regulating and effecting HNS remodeling. PMID- 17412805 TI - Lutropin/choriogonadotropin stimulate the proliferation of primary cultures of rat Leydig cells through a pathway that involves activation of the extracellularly regulated kinase 1/2 cascade. AB - Primary cultures of progenitor and immature rat Leydig cells were established from the testes of 21- and 35-d-old rats, respectively. The cell population remained homogeneous after 4-6 d in culture as judged by staining for 3beta hydroxysteroid dehydrogenase, but the cells were unable to bind 125I-human chorionic gonadotropin (hCG) or to respond to hCG with classical LH receptor (LHR)-mediated responses, including cAMP and inositol phosphate accumulation, steroid biosynthesis, or the phosphorylation of ERK1/2. Infection of primary cultures with recombinant adenovirus coding for beta-galactosidase showed that approximately 65% of the cells are infected. Infection with adenovirus coding for the human LHR (hLHR) allowed for expression of the hLHR at a density of approximately 25,000 receptors per cell and allowed the cells to respond to hCG with increases in cAMP and inositol phosphate accumulation, steroid biosynthesis, and the phosphorylation of ERK1/2. Although progenitor and immature cells were able to respond to hCG with an increase in progesterone, only the immature cells responded with an increase in testosterone. In addition to these classical LHR mediated responses, the primary cultures of progenitor or immature rat Leydig cells expressing the recombinant hLHR proliferated robustly when incubated with hCG, and this proliferative response was sensitive to an inhibitor of ERK1/2 phosphorylation. These studies establish a novel experimental paradigm that can be used to study the proliferative response of Leydig cells to LH/CG. We conclude that activation of the LHR-provoked Leydig cell proliferation requires activation of the ERK1/2 cascade. PMID- 17412806 TI - Constitutive activity of the osteoblast Ca2+-sensing receptor promotes loss of cancellous bone. AB - Changes in extracellular [Ca2+] modulate the function of bone cells in vitro via the extracellular Ca2+-sensing receptor (CaR). Within bone microenvironments, resorption increases extracellular [Ca2+] locally. To determine whether enhanced CaR signaling could modulate remodeling and thereby bone mass in vivo, we generated transgenic mice with a constitutively active mutant CaR (Act-CaR) targeted to their mature osteoblasts by the 3.5 kb osteocalcin promoter. Longitudinal microcomputed tomography of cancellous bone revealed reduced bone volume and density, accompanied by a diminished trabecular network, in the Act CaR mice. The bone loss was secondary to an increased number and activity of osteoclasts, demonstrated by histomorphometry of secondary spongiosa. Histomorphometry, conversely, indicates that bone formation rates were unchanged in the transgenic mice. Constitutive signaling of the CaR in mature osteoblasts resulted in increased expression of RANK-L (receptor activator of nuclear factor kappaB ligand), the major stimulator of osteoclast differentiation and activation, which is the likely underlying mechanism for the bone loss. The phenotype of Act-CaR mice is not attributable to systemic changes in serum [Ca2+] or PTH levels. We provide the first in vivo evidence that increased signaling by the CaR in mature osteoblasts can enhance bone resorption and further propose that fluctuations in the [Ca2+] within the bone microenvironment may modulate remodeling via the CaR. PMID- 17412807 TI - Vasopressin mediates mitogenic responses to adrenalectomy in the rat anterior pituitary. AB - To determine whether increased vasopressinergic activity during chronic stress or adrenalectomy mediates trophic changes in the corticotroph, we examined the effect of peripheral V1 receptor blockade in rats, using the antagonist, dGly[Phaa1,D-tyr(et), Lys, Arg]vasopressin (VP), on the number of pituitary cells taking up bromodeoxyuridine (BrdU) and cells containing immunoreactive ACTH (irACTH). Adrenalectomy significantly increased the number of BrdU- and ACTH labeled cells at 3 and 6 d, and a much larger increase was observed at 28 d. Minipump infusion of V1 antagonist for 28 d, at doses blocking the increases in ACTH and corticosterone induced by exogenous VP, prevented the increases in BrdU incorporation, but not irACTH cells observed 28 d after adrenalectomy. Unexpectedly, colocalization of BrdU with ACTH-positive cells was minor (about three cells per pituitary section), and this was unaffected by adrenalectomy or V1 antagonist infusion. In contrast, adrenalectomy for 6 or 14 d failed to increase BrdU incorporation or irACTH cells in V1b receptor knockout mice while inducing the expected increase in wild-type mice. The data show that VP is required for pituitary mitogenesis after adrenalectomy but, at least in rats, not for increasing the number of corticotrophs. The lack of colocalization of ACTH in mitotic cells suggests that recruitment of corticotrophs during adrenalectomy occurs from undifferentiated cells. PMID- 17412808 TI - Estrogen receptor-beta mediates dihydrotestosterone-induced stimulation of the arginine vasopressin promoter in neuronal cells. AB - Arginine vasopressin (AVP) is a neuropeptide involved in the regulation of fluid balance, stress, circadian rhythms, and social behaviors. In the brain, AVP is tightly regulated by gonadal steroid hormones in discrete regions with gonadectomy abolishing and testosterone replacement restoring normal AVP expression in adult males. Previous studies demonstrated that 17beta-estradiol, a primary metabolite of testosterone, is responsible for restoring most of the AVP expression in the brain after castration. However, 5alpha-dihydrotestosterone (DHT) has also been shown to play a role in the regulation of AVP expression, thus implicating the involvement of both androgen and estrogen receptors (ER). Furthermore, DHT, through its conversion to 5alpha-androstane-3beta,17beta-diol, has been shown to modulate estrogen response element-mediated promoter activity through an ER pathway. The present study addressed two central hypotheses: 1) that androgens directly modulate AVP promoter activity and 2) the effect is mediated by an estrogen or androgen receptor pathway. To that end, we overexpressed androgen receptor, ERbeta, and ERbeta splice variants in a neuronal cell line and measured AVP promoter activity using a firefly luciferase reporter assay. Our results demonstrate that DHT and its metabolite 5alpha-androstane 3beta,17beta-diol stimulate AVP promoter activity through ERbeta in a neuronal cell line. PMID- 17412809 TI - Differential regulation of membrane guanylyl cyclases in congestive heart failure: natriuretic peptide receptor (NPR)-B, Not NPR-A, is the predominant natriuretic peptide receptor in the failing heart. AB - Atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) bind natriuretic peptide receptor (NPR)-A and decrease blood pressure and cardiac hypertrophy by elevating cGMP concentrations. Physiological responses to ANP and BNP are diminished in congestive heart failure (CHF) by an unknown mechanism. C type natriuretic peptide (CNP) binding to NPR-B decreases cardiac hypertrophy, but the effect of CHF on NPR-B is unknown. Here, we measured ANP/NPR-A-dependent and CNP/NPR-B-dependent guanylyl cyclase activities in membranes from failing and nonfailing hearts. Transaortic banding of mice resulted in marked CHF as indicated by increased heart/body weight ratios, increased left ventricular diameters, and decreased ejection fractions. In nonfailed hearts, saturating ANP concentrations increased particulate guanylyl cyclase activity almost 10-fold, whereas saturating CNP concentrations increased activity 6.9-fold, or to about 70% of the ANP response. In contrast, in failed heart preparations, CNP elicited twice as much activity as ANP due to dramatic reductions in NPR-A activity without changes in NPR-B activity. For the first time, these data indicate that NPR-B activity represents a significant and previously unappreciated portion of the natriuretic peptide-dependent guanylyl cyclase activity in the normal heart and that NPR-B accounts for the majority of the natriuretic peptide-dependent activity in the failed heart. Based on these findings, we suggest that drugs that target both NPRs may be more beneficial than drugs like nesiritide (Natrecor) that target NPR-A alone. PMID- 17412810 TI - Bfk, a novel member of the bcl2 gene family, is highly expressed in principal cells of the mouse epididymis and demonstrates a predominant nuclear localization. AB - B-cell lymphoma 2 (BCL2) family kin (BFK) is a recently identified novel protein that is similar to proteins of the BCL2 family. In the present study, we discovered that the mouse Bfk transcript is expressed at the highest level in the epididymis. Two transcripts of 0.9 and 2.6 kb in size were identified, with alternative exon 4 structures, resulting in a difference in the last three to five amino acids of the variants. However, the 0.9-kb transcript was found to be the predominant form in the epididymis and mammary gland, another tissue with strong Bfk expression. Epididymal Bfk expression was regulated both by androgens and other testicular factors. It is thus one of the few initial-segment enriched genes under androgen control, the majority of them being regulated by other testicular factors. BFK protein was expressed specifically in the principal cells of the epididymis. Its nuclear localization was evident in the initial segment and caput epididymis and in the epithelium of pregnant female mammary gland. The expression of BFK-enhanced green fluorescent protein recombinant protein in epididymal cells further confirmed the predominant nuclear localization of BFK with nucleo-cytoplasmic shuttling. Overexpressing BFK in epididymal cells did not induce apoptosis. However, enhanced caspase 3 activation was observed in the presence of BFK upon staurosporine-induced apoptosis. This suggests that BFK may have a proapoptotic role only after the process has been initiated by other mechanisms. Being exceptionally highly expressed in the initial segment, Bfk is suggested to have a role in the differentiation of this segment of the epididymis. PMID- 17412811 TI - Optimization of dosing schedule of daily inhalant dexamethasone to minimize phase shifting of clock gene expression rhythm in the lungs of the asthma mouse model. AB - Glucocorticoid receptor agonists such as dexamethasone (DEXA) have been recommended for the treatment of asthma. An increased frequency of dosing with these drugs seems preferable for cases of severe or uncontrolled asthma. The purpose of this experiment was to find the appropriate dosing schedule (frequency and timing) for DEXA inhalation based on chronotherapeutic dosing to minimize phase shifts of clock function in the lungs of the ovalbumin-treated asthmatic mouse. The daily rhythm of clock gene expression was similar between control and ovalbumin-treated mice. Acute inhalation of DEXA significantly increased mPer1 gene expression in the lungs but not the liver of mice. Daily exposure of DEXA at zeitgeber time 0 (lights on) or at zeitgeber time 18 (6 h after lights off) for 6 d caused a phase advance or phase delay of bioluminescence rhythm in the lungs, respectively, similar to light-induced phase shifts in locomotor activity rhythm. Daily zeitgeber time 0 exposure to DEXA attenuated the expression level of the mClca3 gene, which is associated with mucus overproduction, and there was a phase advancing peak time of the mClca3 rhythm. The present results denote the importance of selecting the most appropriate time of day for nebulizer administration of DEXA to minimize adverse effects such as the phase shifting of clock function in asthmatic lungs. This is the first report of a successful protocol that could obtain phase shifts of clock gene expression rhythm in isolated peripheral organs in vivo. PMID- 17412812 TI - Hepatic branch vagotomy, like insulin replacement, promotes voluntary lard intake in streptozotocin-diabetic rats. AB - Although high insulin concentrations reduce food intake, low insulin concentrations promote lard intake over chow, possibly via an insulin-derived, liver-mediated signal. To investigate the role of the hepatic vagus in voluntary lard intake, streptozotocin-diabetic rats with insulin or vehicle replaced into either the superior mesenteric or jugular veins received a hepatic branch vagotomy (HV) or a sham operation. All rats received a pellet of corticosterone that clamped the circulating steroid at moderately high concentrations to enhance lard intake. After 5 d of recovery, rats were offered the choice of lard and chow for 5 d. In streptozotocin-diabetic rats, HV, like insulin replacement, restored lard intake to nondiabetic levels. Consequently, this reduced chow intake without affecting total caloric intake, and insulin site-specifically increased white adipose tissue weight. HV also ablated the effects of insulin on reducing circulating glucose levels and attenuated the streptozotocin-induced weight loss in most groups. Collectively, these data suggest that the hepatic vagus normally inhibits lard intake and can influence glucose homeostasis and the pattern of white adipose tissue deposition. These actions may be modulated by insulin acting both centrally and peripherally. PMID- 17412813 TI - Autocrine prolactin promotes prostate cancer cell growth via Janus kinase-2 signal transducer and activator of transcription-5a/b signaling pathway. AB - The molecular mechanisms that promote progression of localized prostate cancer to hormone-refractory and disseminated disease are poorly understood. Prolactin (Prl) is a local growth factor produced in high-grade prostate cancer, and exogenously added Prl in tissue or explant cultures of normal and malignant prostate is a strong mitogen and survival factor for prostate epithelium. The key signaling proteins that mediate the biological effects of Prl in prostate cancer are Signal Transducer and Activator of Transcription (Stat)-5a/5b via activation of Janus kinase-2. Importantly, inhibition of Stat5a/b in prostate cancer cells induces apoptotic death. Using a specific Prl receptor antagonist (Delta1-9G129R hPRL), we demonstrate here for the first time that autocrine Prl in androgen independent human prostate cancer cells promotes cell viability via Stat5 signaling pathway. Furthermore, we examined a unique clinical material of human hormone refractory prostate cancers and metastases and show that autocrine Prl is expressed in 54% of hormone-refractory clinical human prostate cancers and 62% prostate cancer metastases. Finally, we demonstrate that autocrine Prl is expressed from both the proximal and distal promoters of the Prl gene in clinical human prostate cancers and in vivo and in vitro human prostate cancer models, independently of pituitary transcription factor-1 (Pit-1). Collectively, the data provide novel evidence for the concept that autocrine Prl signaling pathway is involved in growth of hormone-refractory and metastatic prostate cancer. The study also provides support for the use of Prl receptor antagonists or other therapeutic strategies to block the Prl-Janus kinase-2-Stat5 signaling pathway in advanced prostate cancer. PMID- 17412814 TI - Growth hormone stimulates hepatic expression of bovine growth hormone receptor messenger ribonucleic acid through signal transducer and activator of transcription 5 activation of a major growth hormone receptor gene promoter. AB - The objective of this study was to determine whether and how GH regulates hepatic expression of GH receptor (GHR) mRNA in cattle. Ribonuclease protection assays revealed that injection of GH in a slow-release formula increased both hepatic GHR and IGF-I mRNAs 1 wk after the injection. The increases in GHR and IGF-I mRNAs were highly correlated. Western blot analysis showed that the injection also increased liver GHR protein level. In cattle and other mammals, hepatic GHR mRNA is expressed as variants that differ in the 5'-untranslated region due to the use of different promoters in transcription and/or alternative splicing. We found that GH increased the expression of the liver-specific GHR mRNA variant GHR1A without affecting the other two major GHR mRNA variants in the bovine liver, GHR1B and GHR1C. In transient transfection analyses, GH could robustly activate reporter gene expression from a 2.7-kb GHR1A promoter, suggesting that GH augmentation of GHR1A mRNA expression in the liver is at least partially mediated at the transcriptional level. Additional transfection analyses of serially 5'-truncated fragments of this promoter narrowed the GH-responsive sequence element down to a 210-bp region that contained a putative signal transducer and activator of transcription 5 (STAT5) binding site. EMSAs demonstrated that this putative STAT5 binding site was able to bind to STAT5b protein. In cotransfection assays, deletion of this putative STAT5 binding site abolished most of the GH response of the GHR1A promoter. Like 1-wk GH action, 6-h (i.e. short-term) GH action also increased liver expression of GHR1A and total GHR mRNAs in cattle. These observations together suggest that GH directly stimulates the expression of one GHR mRNA variant, GHR1A, through binding STAT5 to its promoter, thereby increasing GHR mRNA and protein expression in the bovine liver. PMID- 17412815 TI - Increased phosphorylation of myosin light chain prevents in vitro decidualization. AB - Differentiation of stromal cells into decidual cells, which is critical to successful pregnancy, represents a complex transformation requiring changes in cytoskeletal architecture. We demonstrate that in vitro differentiation of human uterine fibroblasts into decidual cells includes down-regulation of alpha-smooth muscle actin and beta-tubulin, phosphorylation of focal adhesion kinase, and redistribution of vinculin. This is accompanied by varied adhesion to fibronectin and a modified ability to migrate. Cytoskeletal organization is determined primarily by actin-myosin II interactions governed by the phosphorylation of myosin light chain (MLC20). Decidualization induced by cAMP [with estradiol 17beta (E) and medroxyprogesterone acetate (P)] results in a 40% decrease in MLC20 phosphorylation and a 55% decline in the long (214 kDa) form of myosin light-chain kinase (MLCK). Destabilization of the cytoskeleton by inhibitors of MLCK (ML-7) or myosin II ATPase (blebbistatin) accelerates decidualization induced by cAMP (with E and P) but inhibits decidualization induced by IL-1beta (with E and P). Adenoviral infection of human uterine fibroblast cells with a constitutively active form of MLCK followed by decidualization stimuli leads to a 30% increase in MLC20 phosphorylation and prevents decidualization. These data provide evidence that the regulation of cytoskeletal dynamics by MLC20 phosphorylation is critical for decidualization. PMID- 17412816 TI - Lipid rafts are triage centers for multimeric and monomeric thyrotropin receptor regulation. AB - The TSH receptor (TSHR), a heptahelical G protein-coupled receptor on the surface of thyrocytes, is a major autoantigen and physiological regulator of the thyroid gland. Unlike other G protein-coupled receptors, the TSHR undergoes posttranslational cleavage of its ectodomain, leading to the existence of several forms of the receptor on the plasma membrane. We previously hypothesized that to achieve high fidelity and specificity of TSH ligand or TSHR autoantibody signaling, the TSHR may compartmentalize into microdomains within the plasma membrane. In support of this hypothesis we have shown previously that TSHRs reside in GM1 ganglioside-enriched lipid rafts in the plasma membrane of TSHR expressing cells. In this study, we further explored the different forms of TSHRs that reside in lipid rafts. We studied both TSHR-transfected cells and rat thyrocytes, using both nondetergent biochemical analyses and receptor-lipid raft colocalization. Using the biochemical approach, we observed that monomeric receptors existed in both raft and nonraft fractions of the cell surface in the steady state. We also demonstrated that the multimeric forms of the receptor were preferentially partitioned into the lipid microdomains. Different TSHR forms, including multimers, were dynamically regulated both by receptor-specific and postreceptor-specific modulators. TSH ligand and TSHR antibody of the stimulating variety induced a decrease of multimeric forms in the raft fractions. In addition, multimeric and monomeric forms of the receptor were both associated with Gsalpha within and without the rafts. Although failure to achieve total lipid raft disruption prevented a conclusion regarding the relative power of TSHR signaling within and without the raft domains, these data showed clearly that not only were a significant proportion of TSHRs residing within lipid microdomains but that constitutive multimerization of TSHRs was actually regulated within the lipid rafts. PMID- 17412817 TI - Estrogen actions on lactotroph proliferation are independent of a paracrine interaction with other pituitary cell types: a study using lactotroph-enriched cells. AB - The mitogenic action of estrogen on estrogen-responsive tissues is suggested to be mediated by paracrine growth factors secreted from neighboring estrogen receptor-positive cells. Using pituitary lactotrophs in primary culture, on which estrogen exerts both mitogenic and antimitogenic actions in a cell context dependent manner, we investigated whether a paracrine cell-to-cell interaction with other pituitary cell types was required for estrogen action. In pituitary cells, enriched for lactotrophs by 85% using differential sedimentation on a discontinuous Percoll gradient, 17beta-estradiol (E2) showed an antimitogenic action on lactotrophs in the presence of IGF-I, which was similar to that in control unenriched cells. Mitogenic actions were also seen in lactotroph-enriched cells when E2 was administered alone, in combination with serum, or in combination with the adenylate cyclase activator forskolin. Similar results were obtained in 90% lactotroph-enriched cells collected by fluorescence-activated cell sorting from transgenic rats expressing enhanced green fluorescent protein under the control of the prolactin promoter. The putative role of basic fibroblast growth factor (bFGF) as a paracrine factor mediating the mitogenic action of estrogen was not supported by the results that: 1) bFGF inhibited lactotroph proliferation; 2) immunoneutralization of bFGF failed to block E2 induced proliferation; and 3) cellular bFGF levels were not altered by E2 treatment. These results suggest that the antimitogenic and mitogenic actions of estrogen on lactotrophs do not require paracrine signals from other pituitary cell types and that estrogen directly influences lactotroph proliferation. PMID- 17412819 TI - Editorial: new vistas on personality disorders and criminal responsibility. PMID- 17412818 TI - Characterization of three growth hormone-responsive transcription factors preferentially expressed in adult female liver. AB - Plasma GH profiles regulate the sexually dimorphic expression of cytochromes P450 and many other genes in rat and mouse liver; however, the proximal transcriptional regulators of these genes are unknown. Presently, we characterize three liver transcription factors that are expressed in adult female rat and mouse liver at levels up to 16-fold [thymus high-mobility group box protein (Tox)], 73-fold [tripartite motif-containing 24 (Trim24)/transcription initiation factor-1alpha (TIF1alpha)], and 125-fold [cut-like 2 (Cutl2)/cut homeobox 2 (Cux2)] higher than in adult males, depending on the strain and species, with Tox expression only detected in mice. In rats, these sex differences first emerged at puberty, when the high prepubertal expression of Cutl2 and Trim24 was extinguished in males but was further increased in females. Rat hepatic expression of Cutl2 and Trim24 was abolished by hypophysectomy and, in the case of Cutl2, was restored to near-female levels by continuous GH replacement. Cutl2 and Trim24 were increased to female-like levels in livers of intact male rats and mice treated with GH continuously (female GH pattern), whereas Tox expression reached only about 40% of adult female levels. Expression of all three genes was also elevated to normal female levels or higher in male mice whose plasma GH profile was feminized secondary to somatostatin gene disruption. Cutl2 and Trim24 both responded to GH infusion in mice within 10-24 h and Tox within 4 d, as compared with at least 4-7 d required for the induced expression of several continuous GH-regulated cytochromes P450 and other female-specific hepatic genes. Cutl2, Trim24, and Tox were substantially up-regulated in livers of male mice deficient in either of two transcription factors implicated in GH regulation of liver sex specificity, namely, signal transducer and activator of transcription 5b (STAT5b) and hepatocyte nuclear factor 4alpha (HNF4alpha), with sex-specific expression being substantially reduced or lost in mice deficient in either nuclear factor. Cutl2 and Trim24 both display transcriptional repressor activity and could thus contribute to the loss of GH-regulated, male-specific liver gene expression seen in male mice deficient in STAT5b or HNF4alpha. Binding sites for Cutl1, whose DNA-binding specificity is close to that of Cutl2, were statistically overrepresented in STAT5b-dependent male-specific mouse genes, lending support to this hypothesis. PMID- 17412820 TI - Intensive case management as a jail diversion program for people with a serious mental illness: a review of the literature. AB - This article reviews the research on intensive case management (ICM) programs as a jail diversion intervention for people with a serious mental illness (SMI). The review includes two types of ICM programs: (a) general ICM programs that included an assessment of arrests and incarceration rates for people with an SMI and (b) ICM programs specifically implemented as a component of a jail diversion intervention for people with an SMI. Results indicate that general ICM programs (19) rarely led to reductions in jail or arrest rates over time, and these rates were similar to those found in standard mental health services. General ICM programs that included an integrated addiction treatment component (8) had mixed results but a trend toward reductions in rates of arrests and incarceration over time for individuals with an SMI and a co-occurring substance use disorder. Results were mixed for jail diversion interventions with an ICM program, but most ICM programs (8) led to significant reductions in arrests and incarcerations over time. Specific elements of effective ICM jail diversion programs are discussed. PMID- 17412821 TI - Identifying subgroups at high risk of dropping out of domestic batterer treatment: the buffering effects of a high school education. AB - This study identifies the subgroups of domestic batterers who are at a low or high risk of failing to complete domestic batterer cognitive behavioral treatment. The sample is composed of 355 domestic batterers ordered to complete treatment, with 31.8% not completing treatment. Three subgroups of batterers were identified as having at least a 60% chance of treatment failure: (a) unemployed generalized aggressors, (b) high school dropouts ordered into substance abuse treatment, and (c) unemployed offenders ordered into substance abuse treatment. Furthermore, a high school education, even when offenders are unemployed or living in poverty, buffers the negative effects of a substance abuse problem among court-mandated batterers required to participate in both domestic violence treatment and substance abuse treatment. Two thirds of substance-abusing high school graduates completed both domestic violence and substance abuse treatment, compared to only one third of the substance-abusing high school dropouts. Implications are discussed. PMID- 17412822 TI - Definitions of and beliefs about wife abuse among undergraduate students of social work. AB - The article focuses on definitions of and beliefs about wife abuse among undergraduate social work students in Israel. Data were collected through self administered questionnaires. The vast majority of students in Study 1 acknowledged acts thought to constitute wife assault and disapproved of a husband's use of force against his wife. The majority of students in Study 2 did not justify wife abuse nor tend to believe that battered women benefit from beating, although they tended to blame the violent husband for his behavior. Significant amounts of the variance in dependent variables were explained by the students' marital role expectations (Study 1) and their attitudes toward women and sex role stereotypes (Study 2). The students' year of study and participation in family violence or wife abuse courses did not contribute toward explaining the variance in their beliefs. Results are discussed in light of the students' patriarchal ideology, and implications for future research are presented. PMID- 17412823 TI - Achieving justice for children: public defenders in Israel's juvenile courts. AB - In 1999, public defender (PD) representation of defendants appearing before Israel's juvenile courts began to be phased in. This article reports some of the major findings of a study that examined the impacts of the introduction of PDs. Analysis of interviews with 14 PDs yielded four major themes concerning the impact of the "arrival" of PDs, nature of the court, PDs' role, and PDs' interactions with other court actors. Analysis of interviews with eight prosecutors yielded seven themes concerning the need for PDs, PD as state agent, PDs' role, harms of legalization, disruption of the court, compromising the therapeutic value of the court hearing, and changes in court process. More generally, both PDs and prosecutors placed uncritical store in the value of rehabilitation alternatives. Indeed, the welfare model continues to shape their roles. The findings can largely be explained in terms of Eisenstein and Jacob's courtroom workgroup model. PMID- 17412824 TI - Family role in the reintegration process of recovering drug addicts: a qualitative review of Israeli offenders. AB - In an outcome evaluation of recovering addicts who participated in a prison-based therapeutic community, 39 clients, a nonrandomized subsample, are interviewed about their rehabilitation and reintegration experience. This study focuses on the family as the main source of support and as an agent of change for the recovering drug addict. Although many studies show marriage and family to be positively related to successful reintegration and rehabilitation, clients in this study report families as having a negative effect on the rehabilitation and reintegration process. This is especially true among those who reunite with their spouse. The findings are discussed in regard to role expectations and support mechanisms, and suggestions are made for further research. PMID- 17412825 TI - Case illustrations of negative sexual experiences among university women in Japan: victimization disclosure and reactions of the confidant. AB - A retrospective study of negative sexual experiences (NSEs) was conducted among 532 Japanese university women. More than one third of the respondents reported having experienced at least one NSE, and two thirds were victimized by strangers. When the woman knew the perpetrator well, the NSE was more likely to be repeated. One sixth of victims did not disclose the incident. Older children disclosed incidents more frequently than did younger children. Women in the total disclosure group disclosed the incidents more quickly than did those in the partial disclosure group. Women victimized by strangers were more likely to disclose the incident completely. Less than 10% of women were disappointed with the reaction of the first-disclosure receiver. Unlike results obtained in previous studies, the victims did not necessarily regard it negatively when the incident was viewed less seriously or they were criticized by first-disclosure receivers. On the other hand, instrumental support was not always supportive. PMID- 17412826 TI - Human organic anion transporter 1B1 and 1B3 function as bidirectional carriers and do not mediate GSH-bile acid cotransport. AB - Organic anion transporting polypeptides (OATP/SLCO) are generally believed to function as electroneutral anion exchangers, but direct evidence for this contention has only been provided for one member of this large family of genes, rat Oatp1a1/Oatp1 (Slco1a1). In contrast, a recent study has indicated that human OATP1B3/OATP-8 (SLCO1B3) functions as a GSH-bile acid cotransporter. The present study examined the transport mechanism and possible GSH requirement of the two members of this protein family that are expressed in relatively high levels in the human liver, OATP1B3/OATP-8 and OATP1B1/OATP-C (SLCO1B1). Uptake of taurocholate in Xenopus laevis oocytes expressing either OATP1B1/OATP-C, OATP1B3/OATP-8, or polymorphic forms of OATP1B3/OATP-8 (namely, S112A and/or M233I) was cis-inhibited by taurocholate and estrone sulfate but was unaffected by GSH. Likewise, taurocholate and estrone sulfate transport were trans stimulated by estrone sulfate and taurocholate but were unaffected by GSH. OATP1B3/OATP-8 also did not mediate GSH efflux or GSH-taurocholate cotransport out of cells, indicating that GSH is not required for transport activity. In addition, estrone sulfate uptake in oocytes microinjected with OATP1B3/OATP-8 or OATP1B1/OATP-C cRNA was unaffected by depolarization of the membrane potential or by changes in pH, suggesting an electroneutral transport mechanism. Overall, these results indicate that OATP1B3/OATP-8 and OATP1B1/OATP-C most likely function as bidirectional facilitated diffusion transporters and that GSH is not a substrate or activator of their transport activity. PMID- 17412827 TI - Hepatocyte growth factor promotes colonic epithelial regeneration via Akt signaling. AB - Hepatocyte growth factor (HGF) can promote the regeneration of injured organs, including HGF gene therapy by electroporation (EP) for liver injury. In this study, we investigated the effect of HGF on dextran sulfate sodium-induced colitis and tried to clarify the regenerative mechanisms of colonic epithelial cells and the signaling pathway involved. Colitis was induced by dextran sulfate sodium in mice, together with HGF gene transfer by EP. On day 10, the colitis was evaluated histologically and by Western blot analysis. The colonic epithelial cell line MCE301 was exposed to HGF protein, and its proliferation and activated signaling pathway were analyzed. In vivo, the histological score improved and the number of Ki-67-positive epithelial cells increased in the HGF-treated mice compared with the controls. Western blot analysis showed enhanced expression of phospho-Akt in the HGF-treated mice compared with the controls. In vitro, HGF stimulated the proliferation of MCE301 cells. There was enhanced phospho-Akt expression for more than 48 h after HGF stimulation, although phospho-ERK1/2 was enhanced for only 10 min. LY-294002 or Akt small interfering RNA suppressed cell proliferation induced by HGF. Thus HGF induces the proliferation of colonic epithelial cells via the phosphatidylinositol 3-kinase/Akt signaling pathway. HGF gene therapy can attenuate acute colitis via epithelial cell proliferation through the PI3K/Akt pathway. These data suggested that HGF gene therapy by EP may be effective for the regeneration and repair of injured epithelial cells in inflammatory bowel disease. PMID- 17412828 TI - Human cecal bile acids: concentration and spectrum. AB - To obtain information on the concentration and spectrum of bile acids in human cecal content, samples were obtained from 19 persons who had died an unnatural death from causes such as trauma, homicide, suicide, or drug overdose. Bile acid concentration was measured via an enzymatic assay for 3alpha-hydroxy bile acids; bile acid classes were determined by electrospray ionization mass spectrometry and individual bile acids by gas chromatography mass spectrometry and liquid chromatography mass spectrometry. The 3alpha-hydroxy bile acid concentration (mumol bile acid/ml cecal content) was 0.4 +/- 0.2 mM (mean +/- SD); the total 3 hydroxy bile acid concentration was 0.6 +/- 0.3 mM. The aqueous concentration of bile acids (supernatant after centrifugation) was identical, indicating that most bile acids were in solution. By liquid chromatography mass spectrometry, bile acids were mostly in unconjugated form (90 +/- 9%, mean +/- SD); sulfated, nonamidated bile acids were 7 +/- 5%, and nonsulfated amidated bile acids (glycine or taurine conjugates) were 3 +/- 7%. By gas chromatography mass spectrometry, 10 bile acids were identified: deoxycholic (34 +/- 16%), lithocholic (26 +/- 10%), and ursodeoxycholic (6 +/- 9), as well as their primary bile acid precursors cholic (6 +/- 9%) and chenodeoxycholic acid (7 +/- 8%). In addition, 3beta-hydroxy derivatives of some or all of these bile acids were present and averaged 27 +/- 18% of total bile acids, indicating that 3beta hydroxy bile acids are normal constituents of cecal content. In the human cecum, deconjugation and dehydroxylation of bile acids are nearly complete, resulting in most bile acids being in unconjugated form at submicellar and subsecretory concentrations. PMID- 17412829 TI - Pharmacokinetic and pharmacodynamic properties of inhaled ciclesonide. AB - Inhaled corticosteroids are recommended first-line therapy for persistent asthma of all severities; however, oropharyngeal and systemic adverse events can be a concern. Inhaled corticosteroids exert their therapeutic and adverse effects by interacting with glucocorticoid receptors within and outside the lungs, respectively. Ciclesonide is a novel inhaled corticosteroid that possesses a unique pharmacokinetic and pharmacodynamic profile. Ciclesonide is inactive itself and converted to its pharmacologically active metabolite, desisobutyryl ciclesonide, in the target organ, the lungs. Pulmonary activation combined with low oral deposition may minimize oropharyngeal adverse events, and low oral bioavailability, rapid clearance, and high protein binding may reduce systemic exposure. In addition, high pulmonary deposition due to the highly respirable particles, combined with the potential for prolonged lung retention via lipid conjugation, provides for effective therapeutic action. PMID- 17412830 TI - Projected impacts of climate change on salmon habitat restoration. AB - Throughout the world, efforts are under way to restore watersheds, but restoration planning rarely accounts for future climate change. Using a series of linked models of climate, land cover, hydrology, and salmon population dynamics, we investigated the impacts of climate change on the effectiveness of proposed habitat restoration efforts designed to recover depleted Chinook salmon populations in a Pacific Northwest river basin. Model results indicate a large negative impact of climate change on freshwater salmon habitat. Habitat restoration and protection can help to mitigate these effects and may allow populations to increase in the face of climate change. The habitat deterioration associated with climate change will, however, make salmon recovery targets much more difficult to attain. Because the negative impacts of climate change in this basin are projected to be most pronounced in relatively pristine, high-elevation streams where little restoration is possible, climate change and habitat restoration together are likely to cause a spatial shift in salmon abundance. River basins that span the current snow line appear especially vulnerable to climate change, and salmon recovery plans that enhance lower-elevation habitats are likely to be more successful over the next 50 years than those that target the higher-elevation basins likely to experience the greatest snow-rain transition. PMID- 17412831 TI - H/ACA small nucleolar RNA pseudouridylation pockets bind substrate RNA to form three-way junctions that position the target U for modification. AB - During the biogenesis of eukaryotic ribosomal RNA (rRNA) and spliceosomal small nuclear RNA (snRNA), uridines at specific sites are converted to pseudouridines by H/ACA ribonucleoprotein particles (RNPs). Each H/ACA RNP contains a substrate specific H/ACA RNA and four common proteins, the pseudouridine synthase Cbf5, Nop10, Gar1, and Nhp2. The H/ACA RNA contains at least one pseudouridylation (psi) pocket, which is complementary to the sequences flanking the target uridine. In this article, we show structural evidence that the psi pocket can form the predicted base pairs with substrate RNA in the absence of protein components. We report the solution structure of the complex between an RNA hairpin derived from the 3' psi pocket of human U65 H/ACA small nucleolar RNA (snoRNA) and the substrate rRNA. The snoRNA-rRNA substrate complex has a unique structure with two offset parallel pairs of stacked helices and two unusual intermolecular three-way junctions, which together organize the substrate for docking into the active site of Cbf5. The substrate RNA interacts on one face of the snoRNA in the complex, forming a structure that easily could be accommodated in the H/ACA RNP, and explains how successive substrate RNAs could be loaded onto and unloaded from the H/ACA RNA in the RNP. PMID- 17412832 TI - Three functional variants of IFN regulatory factor 5 (IRF5) define risk and protective haplotypes for human lupus. AB - Systematic genome-wide studies to map genomic regions associated with human diseases are becoming more practical. Increasingly, efforts will be focused on the identification of the specific functional variants responsible for the disease. The challenges of identifying causal variants include the need for complete ascertainment of genetic variants and the need to consider the possibility of multiple causal alleles. We recently reported that risk of systemic lupus erythematosus (SLE) is strongly associated with a common SNP in IFN regulatory factor 5 (IRF5), and that this variant altered spicing in a way that might provide a functional explanation for the reproducible association to SLE risk. Here, by resequencing and genotyping in patients with SLE, we find evidence for three functional alleles of IRF5: the previously described exon 1B splice site variant, a 30-bp in-frame insertion/deletion variant of exon 6 that alters a proline-, glutamic acid-, serine- and threonine-rich domain region, and a variant in a conserved polyA+ signal sequence that alters the length of the 3' UTR and stability of IRF5 mRNAs. Haplotypes of these three variants define at least three distinct levels of risk to SLE. Understanding how combinations of variants influence IRF5 function may offer etiological and therapeutic insights in SLE; more generally, IRF5 and SLE illustrates how multiple common variants of the same gene can together influence risk of common disease. PMID- 17412833 TI - Oscillation patterns in negative feedback loops. AB - Organisms are equipped with regulatory systems that display a variety of dynamical behavior ranging from simple stable steady states, to switching and multistability, to oscillations. Earlier work has shown that oscillations in protein concentrations or gene expression levels are related to the presence of at least one negative feedback loop in the regulatory network. Here, we study the dynamics of a very general class of negative feedback loops. Our main result is that, when a single negative feedback loop dominates the dynamical behavior, the sequence of maxima and minima of the concentrations exhibit a pattern that uniquely identifies the interactions of the loop. This allows us to devise an algorithm to (i) test whether observed oscillating time series are consistent with a single underlying negative feedback loop, and if so, (ii) reconstruct the precise structure of the loop, i.e., the activating/repressing nature of each interaction. This method applies even when some variables are missing from the data set, or if the time series shows transients, like damped oscillations. We illustrate the relevance and the limits of validity of our method with three examples: p53-Mdm2 oscillations, circadian gene expression in cyanobacteria, and cyclic binding of cofactors at the estrogen-sensitive pS2 promoter. PMID- 17412834 TI - Structural and dynamic properties of propane coordinated to TpRh(CNR) from a confrontation between theory and experiment. AB - Density functional calculations with the B3PW91 functional have been carried out on the TpRh(CNMe) species [Tp = HB(pyrazolyl)(3)] as a model for Tp'Rh(CNCH(2)CMe(3)) [Tp' = HB(3,5-dimethylpyrazolyl)(3)] in interaction with propane. Two sigma complexes have been found as minima coordinated through either a methyl or a methylene C H bond, the former being more stable. The approach of the alkane to TpRh(CNMe) has been studied. Although no transition state could be located, study of this path reveals the key importance of the partial decoordination of one pyrazole ring. The full coordination of the alkane can only be achieved when the metal is essentially in a square pyramid coordination with one of the three pyrazole groups only weakly interacting with Rh. The main reaction of the methyl sigma complex is oxidative addition, leading to the n propyl hydride complex. In contrast, two reactions are found for the methylene sigma complex: (i) oxidative addition to give the isopropyl complex and (ii) exchange between the secondary and primary C H bonds to convert the methylene complex of propane into a methyl complex of propane. This latter reaction has a much lower barrier than the oxidative addition at the methylene C H bond. The results account well for most of the experimental results obtained from kinetic studies. Steric factors are found to control the energy barriers between these various processes, disfavoring any process that brings the central carbon into close proximity to Rh. PMID- 17412835 TI - Daam1 regulates the endocytosis of EphB during the convergent extension of the zebrafish notochord. AB - Convergent extension (CE) movement of cells is one of the fundamental processes that control the organized morphogenesis of tissues and organs. The molecular events connecting the noncanonical Wnt pathway and CE movement, however, are not well understood. We show that subcellular localization of Daam1, an essential component of noncanonical Wnt signaling, changes dynamically during notochord formation. In the early phases, Daam1 complexes with EphB receptors and Disheveled 2. This complex is incorporated into endocytic vesicles in a dynamin dependent manner, thereby resulting in the removal of EphB from the cell surface with subsequent switching of cell adhesiveness. In the next step, Daam1 colocalizes with the actin cytoskeleton to induce morphological extension of cells. We elucidate the molecular mechanism underlying the CE movement of notochord cells with Daam1 as a dynamic coordinator of endocytosis and cytoskeletal remodeling. PMID- 17412836 TI - Dynamin 2 is required for the enhancement of HIV-1 infectivity by Nef. AB - Nef is a virulence factor of HIV-1 and other primate lentiviruses that is crucial for rapid progression to AIDS. In cell culture, Nef increases the infectivity of HIV-1 progeny virions by an unknown mechanism. We now show that dynamin 2 (Dyn2), a key regulator of vesicular trafficking, is a binding partner of Nef that is required for its ability to increase viral infectivity. Dominant-negative Dyn2 or the depletion of Dyn2 by small interfering RNA potently inhibited the effect of Nef on HIV-1 infectivity. Furthermore, in Dyn2-depleted cells, this function of Nef could be rescued by ectopically expressed Dyn2 but not by Dyn1, a closely related isoform that does not bind Nef. The infectivity enhancement by Nef also depended on clathrin, because it was diminished in clathrin-depleted cells and profoundly inhibited in cells expressing the clathrin-binding domain of AP180, which blocks clathrin-coated pit formation but not clathrin-independent endocytosis. Together, these findings imply that the infectivity enhancement activity of Nef depends on Dyn2- and clathrin-mediated membrane invagination events. PMID- 17412837 TI - Characterizing tumor-promoting T cells in chemically induced cutaneous carcinogenesis. AB - There is a longstanding but poorly understood epidemiologic link between inflammation and cancer. Consistent with this, we previously showed that alphabeta T cell deficiency can increase resistance to chemical carcinogenesis initiated by 7,12-dimethylbenz[a]anthracene and promoted by phorbol 12-myristate 13-acetate. This provoked the hypothesis that alphabeta T cell deficiency removed T regulatory cells that limit the anti-tumor response or removed a specific tumor promoting (T-pro) T cell population. Here we provide evidence for the latter, identifying a novel CD8(+) subset that is a candidate for T-pro cells. We demonstrate that CD8 cell-deficient mice show substantially less tumor incidence and progression to carcinoma, whereas susceptibility is restored by CD8(+) cell reconstitution. To characterize the putative T-pro cells, tumor-infiltrating lymphocytes were isolated from normal and CD4(-/-) mice, revealing an activated population of T cell receptor alphabeta(+)CD8(+)CD44(+)CD62L(-) cells expressing the inflammatory mediators IFNgamma, TNFalpha, and cyclooxygenase-2, but deficient in perforin, relative to recirculating cells of equivalent phenotype. This novel population of CD8(+) T cells has intriguing similarities with other lymphocytes that have been associated with tissue growth and invasiveness and has implications for inflammation-associated carcinogenesis, models of cancer immunosurveillance, and immunotherapeutic strategies. PMID- 17412838 TI - Pioglitazone and rosiglitazone decrease prostaglandin E2 in non-small-cell lung cancer cells by up-regulating 15-hydroxyprostaglandin dehydrogenase. AB - Lung cancer cells elaborate the immunosuppressive and antiapoptotic mediator prostaglandin E(2) (PGE(2)), a product of cyclooxygenase-2 (COX-2) enzyme activity. Because peroxisome proliferator-activated receptor (PPAR)gamma ligands, such as thiazolidinediones (TZDs), inhibit lung cancer cell growth, we examined the effect of the TZDs pioglitazone and rosiglitazone on PGE(2) levels in non small-cell lung cancer (NSCLC) A427 and A549 cells. Both TZDs inhibited PGE(2) production in NSCLC cells via a COX-2 independent pathway. To define the mechanism underlying COX-2 independent suppression of PGE(2) production, we focused on other enzymes responsible for the synthesis and degradation of PGE(2). The expression of none of the three prostaglandin synthases (microsomal PGES1, PGES2 and cystosolic PGES) was down-regulated by the TZDs. It is noteworthy that 15-hydroxyprostaglandin dehydrogenase (15-PGDH), an enzyme that produces biologically inactive 15-ketoprostaglandins from active PGE(2), was induced by TZDs. The TZD-mediated suppression of PGE(2) concentration was significantly inhibited by small interfering RNA to 15-PGDH. Studies using dominant-negative PPARgamma overexpression or 2-chloro-5-nitrobenzanilide (GW9662; a PPARgamma antagonist) revealed that the suppressive effect of the TZDs on PGE(2) is PPARgamma-independent. Together, these findings indicate that it is possible to use a clinically available pharmacological intervention to suppress tumor-derived PGE(2) by enhancing catabolism rather than blocking synthesis. PMID- 17412839 TI - Who receives long-acting antipsychotic medications? PMID- 17412840 TI - Economic grand rounds: A pay-for-performance program for behavioral health care practitioners. AB - This column describes a pay-for-performance program for behavioral health care practitioners. Implemented in 1996 by a large national health insurer, the program's goals are to improve the quality of care, recognize the practitioners who provide higher-quality care, demonstrate the value of behavioral health services to purchasers, and help providers align their practices with national standards. A future goal is to provide patients with data on provider quality to improve their treatment decisions. Important considerations in measure development include application of the measure to all disciplines, feasible data collection processes for providers, creation of clinically meaningful and fair measures, and selection of measures with large baseline variability. PMID- 17412841 TI - State mental health policy: Promoting legislation and public policy debates in state legislatures: a psychiatrist's perspective. AB - Psychiatrists are urged to get involved in promoting legislation and public policy debates in state legislatures to effectively advocate for positive change in legislation and policy making. This column focuses on strategies that New Hampshire Psychiatric Society members have found effective in engaging policy makers and legislators in a dialogue that assertively promotes the views of patients with mental illness and the profession of psychiatry. PMID- 17412842 TI - Best practices: The development of the Social Cognition and Interaction Training program for schizophrenia spectrum disorders. AB - This column describes the development of a treatment, the Social Cognition and Interaction Training (SCIT) program, a group-based intervention delivered weekly over a six-month period, with the purpose of improving both social cognition and social functioning among persons with schizophrenia spectrum disorders. SCIT comprises three phases: emotion training, figuring out situations, and integration. Initial pilot testing of 17 inpatients showed that SCIT was associated with improved emotion perception, improved theory of mind, and a reduced tendency to attribute hostile intent to others, with effect sizes being in the medium-large range. Although research is still in the early phases, SCIT is a potential best practice. PMID- 17412843 TI - Best practices: Utilizing risk-of-violence findings for continuity of care. AB - Best practices are recommended for assessing risk of violence and utilizing those findings to communicate with other mental health providers to manage the potential for violence as clients are transferred or referred. These recommendations are the fruition of a five-year collaboration of the Forensic Risk Assessment Work Group of the Ohio Department of Mental Health's statewide Forensic Product Line Leadership Team. Accurate assessment of static and dynamic risk factors, effective communication of risk of violence, incorporation of dynamic risk factors into the treatment planning process, and implementation of evidence-based risk-management practices are all emphasized as best practices. PMID- 17412844 TI - Best practices: A systematic approach to the management of patients who refuse medications in an assertive community treatment team setting. AB - A significant proportion of patients of assertive community treatment (ACT) teams will adamantly refuse medication. Whether the team should continue to encourage medication or put a hold on advocating for medication is a clinical and ethical dilemma. On the basis of their clinical experiences, the authors propose best practices criteria that ACT teams can consider in deciding whether medications may be temporarily discontinued when a patient refuses them. The authors suggest that in some circumstances stopping medications in such a case may help in the development or repair of a therapeutic alliance over the long term. PMID- 17412845 TI - Law & psychiatry: The new lie detectors: neuroscience, deception, and the courts. AB - This column examines the use of two technologies in lie detection. "Brain fingerprinting" is based on the finding that the brain generates a unique brain wave pattern when a person encounters a familiar stimulus. Use of functional magnetic resonance imaging in lie detection derives from studies suggesting that persons asked to lie show different patterns of brain activity than they do when being truthful. Issues related to the use of such evidence in courts are discussed. The author concludes that neither approach is currently supported by enough data regarding its accuracy in detecting deception to warrant use in court. PMID- 17412846 TI - Personal accounts: My sadness within. PMID- 17412847 TI - Connectedness and citizenship: redefining social integration. AB - OBJECTIVE: Despite decades of deinstitutionalization, individuals with psychiatric disabilities living outside the hospital may be described as in the community, but not of it. To effectively address the persisting problem of social exclusion of persons with psychiatric disabilities, new conceptual tools are needed. To address this need, a new definition of social integration is offered. METHODS: The definition is based on data from a qualitative study. Data collection consisted of individual, unstructured interviews with 56 adults who have been psychiatrically disabled (N=78 interviews) as well as ethnographic visits to five service sites working to promote social integration for their users (N=8 visits). An interpretive approach was used to analyze the data. RESULTS: Social integration is defined as a process, unfolding over time, through which individuals who have been psychiatrically disabled increasingly develop and exercise their capacities for connectedness and citizenship. Connectedness denotes the construction and successful maintenance of reciprocal interpersonal relationships. Social, moral, and emotional competencies are required to sustain connectedness. Citizenship refers to the rights and privileges enjoyed by members of a democratic society and to the responsibilities these rights engender. The definition calls for full rights and responsibilities of citizenship. CONCLUSIONS: The new definition sets an ideal, but not unrealistic, standard for social integration in the context of psychiatric disability. High standards encourage mental health professionals and policy makers to rethink what is possible for mental health services and to raise expectations for connectedness and citizenship among persons once disabled by mental illness. PMID- 17412848 TI - Barriers to the appropriate clinical use of medications that improve the cognitive deficits of schizophrenia. AB - A high priority has been placed on developing medications to treat the cognitive deficits associated with schizophrenia, but less attention has been given to planning for appropriate use of these medications in practice. The cognitive deficits of schizophrenia present several complexities as a treatment target that may limit a clinician's ability to prescribe cognitive enhancers to the patients who need them and to monitor for improvements in outcomes. In this review the neuropsychological evidence regarding cognition and functioning is discussed with a view toward how this evidence might guide clinicians' prescribing practices. Three challenges regarding the use of cognitive enhancers in schizophrenia are discussed. First, laboratory constructs of cognition are not equivalent to cognitive skills and behaviors seen in the clinic. The evidence generated in clinical trials of cognitive enhancers may have uncertain ecological validity. Second, objective scores on cognitive tests often do not match clinicians' and patients' perceptions of cognitive deficits. Mismatch between objective and subjective assessments of cognition may complicate the monitoring of medication. Third, although reductions in disability are desired outcomes of cognitive enhancement, clinicians may not be able to rely on assessments of patients' functional status to determine whether cognition medications are effective. In summary, data on the clinical meaning of neuropsychological constructs, careful selection of outcome measures for randomized clinical trials, and effectiveness trials could help ensure that cognition-enhancing medications can be appropriately prescribed in usual practice settings to the patients who can benefit from them. PMID- 17412849 TI - Characteristics and use patterns of patients taking first-generation depot antipsychotics or oral antipsychotics for schizophrenia. AB - OBJECTIVE: Investigators compared patient characteristics and antipsychotic use patterns between individuals with schizophrenia treated in usual care with first generation depot antipsychotics and those treated with oral antipsychotics (first or second-generation or both). METHODS: Analyses used data from the U.S. Schizophrenia Care and Assessment Program, a large, prospective study of treatment for schizophrenia conducted July 1997 through September 2003. Participants were assessed at enrollment and every six months thereafter with patient self-report, validated psychiatric measures, and systematic extraction of medical records. Individuals treated with a first-generation depot antipsychotic at any time during the three-year study (N=569) were compared with those treated with only oral antipsychotics (N=1,617) on characteristics at enrollment and medication use pattern during the year after enrollment. RESULTS: Compared with patients receiving only oral antipsychotics, participants treated with depot medications (haloperidol or fluphenazine decanoate) were more likely to be African American (p<.001); less likely to be a veteran (p=.005); had more psychiatric hospitalizations in the year before enrollment (p<.001); and were more likely to have been arrested (p<.001), to use alcohol and illicit substances (p<.001), and to show higher psychopathology, particularly psychotic symptoms and disorganized thinking (p<.01 for both). In the year after enrollment, participants treated with depot medications had a high mean medication possession ratio (91%), and most of the medication regimens (68%) were augmented with oral antipsychotics for prolonged durations (median of 144 days). CONCLUSIONS: Patients with schizophrenia treated with first-generation depot antipsychotics differed from those treated with only oral antipsychotics. Findings suggest that first-generation depot antipsychotics might address some unmet needs of a unique subgroup of patients with schizophrenia. PMID- 17412850 TI - Long-term antipsychotic polypharmacy in the VA health system: patient characteristics and treatment patterns. AB - OBJECTIVE: Although antipsychotic polypharmacy is being prescribed with increasing frequency, few studies have described patient characteristics and treatment patterns associated with long-term use of this treatment strategy. METHODS: By using data from the National Psychosis Registry of the Department of Veterans Affairs, 5,826 patients with schizophrenia or schizoaffective disorder who received long-term antipsychotic polypharmacy (simultaneous treatment with two or more antipsychotics for 90 or more days) during fiscal year 2000 and 39,745 patients who received long-term antipsychotic monotherapy were identified. By using multivariate regression models, patient demographic and clinical characteristics, antipsychotic dosages, and use of antiparkinson and adjunctive psychotropic medications were compared between the two groups. RESULTS: Patients were more likely to receive antipsychotic polypharmacy if they were younger, were unmarried, had a military service-connected disability, had schizophrenia rather than schizoaffective disorder, or had greater use of inpatient and outpatient mental health services. Patients were less likely to receive antipsychotic polypharmacy if they were African American, had concurrent diagnoses of depression or substance use disorder, or had greater medical comorbidity. For most antipsychotics, dosages prescribed for patients receiving polypharmacy were the same or modestly higher than those prescribed for patients receiving monotherapy. Patients given prescriptions for polypharmacy were more likely to receive antiparkinson medications, antianxiety agents, and mood stabilizers and equally likely to receive concurrent treatment with antidepressants. CONCLUSIONS: Long-term antipsychotic polypharmacy appears to be reserved for more severely ill patients with psychotic symptoms rather than mood symptoms. These patients may experience increased adverse effects as a result of excess antipsychotic exposure. PMID- 17412851 TI - Analysis of U.S. trends in discharges from general hospitals for episodes of serious mental illness, 1995-2002. AB - OBJECTIVE: The aim of this study was to ascertain trends from 1995 to 2002 in general hospital discharges among adults in the United States with serious mental illness. METHODS: As of 2002 the National Hospital Discharge Survey collected data on approximately 327,000 discharges from 445 hospitals. ICD-9-CM codes were used to identify the discharges between 1995 and 2002 associated with serious mental illness. RESULTS: Annual discharge rates involving serious mental illness increased by 34.7%, from 29.1 discharges per 10,000 in the U.S. adult population (18 years and older) in 1995 to 39.2 discharges per 10,000 in the U.S. adult population in 2002. Hospital discharges involving serious mental illness increased significantly in the black population and among young adults. For adults of ages 18 to 24, discharges per 10,000 increased from 19.9 in 1995 to 42.3 in 2002. A substantial increasing trend was seen for the Northeast and South census regions. There was an increase in the proportion of discharges associated with hospitalizations for serious mental illness that were covered by private payers, whereas there was a significant decline in such discharges when patients were covered by government programs. CONCLUSIONS: The increasing trend in general hospital discharges involving serious mental illness has continued into recent years. Further investigations are needed to understand how patient- and system level factors have contributed to the increasing trend in general hospital discharges involving an episode of serious mental illness. PMID- 17412852 TI - Rural-urban differences in hospitalization rates of primary care patients with depression. AB - OBJECTIVE: Individuals with depression in rural areas showed greater odds of hospitalization over one year than their urban counterparts in a single-state study; however, differences disappeared in models controlling for recent outpatient specialty care. To investigate whether these results are generalizable to a broader geographic area, the research team analyzed an 11-state database to test whether rural primary care patients with depression had greater odds of hospitalization over two years for physical and emotional problems, compared with their urban counterparts. METHODS: Hypotheses were tested by conducting a preplanned secondary analysis of data for 1,455 patients with depression in the Quality Improvement for Depression (QID) database. This database was developed in a two-year cooperative trial that evaluated quality initiatives to improve primary care depression treatment. QID studies, including Partners in Care and Quality Enhancement by Strategic Teaming, recruited patients from rural and urban areas. RESULTS: Multivariate analyses demonstrated that compared with their urban counterparts, rural patients with depression had significantly higher odds of being hospitalized for physical problems (13% versus 7%, OR=1.8, 95% confidence interval [CI]=1.2-2.8, p<.01 at six months) and for emotional problems (4% versus 2%, OR=2.3, CI=1.0-5.4, p=.05 at 18 months). Hospitalization differences were not reduced in models controlling for outpatient specialty care in the previous six months. CONCLUSIONS: Although national studies report that all-cause hospitalization rates are comparable for rural and urban populations, rural patients with depression in this 11-site study had greater odds of hospitalization for both physical and emotional problems over two years, compared with their urban counterparts, suggesting that the potential for reducing hospitalization rates among rural patients with depression should be addressed by depression care management programs serving this population. PMID- 17412853 TI - Psychiatric disorders among adults seeking emergency disaster assistance after a wildland-urban interface fire. AB - OBJECTIVE: This study estimated the prevalence of psychopathology at a three month follow-up among persons seeking emergency relief services after a wildfire and identified a practical screener for use in these disaster assistance settings to aid early identification of persons at risk of subsequent psychopathology. METHODS: During the October 2003 California firestorm that occurred at the wildland-urban interface, 357 persons who were seeking assistance from adjacent American Red Cross and government relief centers were recruited for this study. Within days of mandatory evacuation, participants completed baseline self administered questionnaires assessing demographic characteristics, initial subjective reactions, and degree of fire exposure. At the three-month follow-up, symptoms of posttraumatic stress disorder (PTSD) and major depression were measured via a mailed survey. RESULTS: At follow-up 33% showed evidence of probable major depression; 24% exhibited probable PTSD. On a bivariate basis, seven initial reaction and fire exposure items were significantly associated with subsequent psychopathology. Best-subsets logistic regression analyses revealed that property damage and physical injury were the best multivariate predictors of psychopathology at follow-up. No additional items provided a significant incremental improvement in prediction. CONCLUSIONS: Individuals seeking immediate emergency assistance related to the wildland-urban interface fire were at elevated risk of psychopathology in the weeks after the fire. A short, easily administered, two-item screener, composed of items assessing fire exposure severity, appears to hold promise for aiding early identification of persons at risk of postfire psychopathology. These findings may also have implications for other mass disasters. PMID- 17412854 TI - Gender differences in the annual income of psychiatrists. AB - OBJECTIVE: Female physicians have traditionally earned less than their male counterparts, even after adjustment for specialty, workload, and physician characteristics. In the 1980s female psychiatrists earned incomes that were 13% lower than those of their male counterparts; however, important explanatory variables for such differences were not incorporated into that analysis. This study examined whether a gender income gap among psychiatrists persisted in the 1990s. METHODS: Survey responses were used that were obtained between 1992 and 2001 from 976 actively practicing white psychiatrists (weighted N=941). Because of low numbers of black respondents to the surveys, we were unable to examine the influence of race on psychiatrists' incomes. Linear regression modeling was performed to determine the association between gender and annual incomes after controlling for workload, provider characteristics, and practice characteristics. RESULTS: Among white psychiatrists, women reported working 13% fewer annual hours than their male counterparts, and women had practiced medicine for fewer years than men. Also, women were more likely to be employees of the practice, as opposed to having an ownership interest in the practice, and were more likely to be board certified, although these findings were not significant. After adjustment for workload, provider characteristics, and practice characteristics, the mean annual income for women was $140,615, or $31,962 (19%) lower than that of men (95% confidence interval of $41,834-$22,090 lower, p<.001). CONCLUSIONS: During the 1990s female gender was associated with lower annual incomes among white psychiatrists; compared with previous reports from earlier periods the income disparity appears to be widening. These findings warrant further exploration. PMID- 17412855 TI - Categorization of aggressive acts committed by chronically assaultive state hospital patients. AB - OBJECTIVE: This study examined factors motivating inpatient aggression in a sample of chronically assaultive state hospital patients. METHODS: Inpatients who had committed three or more assaults over a one-year period were identified by using an incident report database. Aggressive episodes were categorized as impulsive, organized, or psychotic by using a procedure for classifying assaultive acts based on record review. Each assault type was further subcategorized. The relationship between assault type, victim (staff or patient), and legal status of the assaulter was also assessed. RESULTS: A total of 839 assaults committed by 88 chronically aggressive patients were reviewed. Although most patients had a primary psychotic disorder, the most common type of assault was impulsive (54%), rather than psychotic or organized. Staff were most often victimized by impulsive assaults in situations involving attempts to change a patient's unwanted behavior and refusal of a patient request. Organized and psychotic assaults occurred less frequently (29% and 17%, respectively) and were more likely to target other patients. Organized assaults were most often motivated by a desire to seek revenge. Psychotic assaults were most often committed by an assailant acting under the influence of paranoid ideations. Civilly committed patients were overrepresented in the sample. Criminally committed patients committed more acts of organized aggression, although this finding did not reach significance. CONCLUSIONS: These findings indicate that assaultive behavior among state hospital inpatients is complex and heterogeneous. Because each type of assault requires a different management approach, characterizing aggressive behavior may be important in determining which institutional programs and treatment-plan interventions to implement when addressing inpatient aggression. PMID- 17412856 TI - The MIRECC version of the Global Assessment of Functioning scale: reliability and validity. AB - OBJECTIVE: This study examined the reliability and convergent, discriminant, and predictive validity of the Mental Illness Research, Education, and Clinical Center (MIRECC) version of the Global Assessment of Functioning (GAF) scale. The MIRECC GAF measures occupational functioning, social functioning, and symptom severity on three subscales. METHODS: MIRECC GAF ratings were obtained for 398 individuals with schizophrenia or schizoaffective disorder who were receiving treatment at three Veterans Affairs mental health clinics. Assessments were completed by using the Positive and Negative Syndrome Scale and the Quality of Life Interview at baseline and nine months later. RESULTS: All three MIRECC GAF subscales exhibited very high levels of reliability. The occupational and symptom subscales showed good convergent and discriminant validity. The social subscale was related to measures of social functioning and, to a greater degree, symptom severity. The occupational and social subscales significantly predicted their respective domains at the nine-month follow-up. The symptom subscale predicted negative symptoms at follow-up; however, it did not predict positive symptoms or cognitive disorientation. Instead, the social subscale was predictive of cognitive disorientation at follow-up. When the standard GAF was routinely administered by clinicians, scores demonstrated little validity. CONCLUSIONS: The three MIRECC GAF subscales can be scored reliably, and they have good concurrent and predictive validity. Further work is needed on brief measures of patient functioning, especially measures of social functioning. PMID- 17412857 TI - Quality of diabetes care among adults with serious mental illness. AB - OBJECTIVE: The study compared the quality of care for type 2 diabetes delivered to two groups with type 2 diabetes--adults with serious mental illness and those with no serious mental illness--in a range of community-based clinic settings. METHODS: Cross-sectional analyses of medical chart data from 300 patients (201 with serious mental illness and 99 without serious mental illness) were used to examine indicators of the quality of care established by the Diabetes Quality Improvement Project. Recommended services assessed included glycosylated hemoglobin examination, eye and foot examinations, blood pressure check, and urine and lipid profiles. Self-report data were used to compare receipt of provider-delivered diabetes education and receipt of cues regarding self management of diabetes for the two study groups. RESULTS: Evidence of lower quality of diabetes care was found for persons with serious mental illness as reflected by their receipt of fewer recommended services and less education about diabetes, compared with those without serious mental illness. Although participants with serious mental illness were less likely to receive cues from providers regarding the need for glucose self-monitoring, they were as likely as those without serious mental illness to receive cues regarding diet and medication adherence. CONCLUSIONS: Although participants with serious mental illness received some services that are indicated in quality-of-care standards for diabetes, they were less likely to receive the full complement of recommended services and care support, suggesting that more effort may be required to provide optimal diabetes care to these vulnerable patients. PMID- 17412858 TI - Outcomes of obese, clozapine-treated inpatients with schizophrenia placed on a six-month diet and physical activity program. AB - OBJECTIVE: Patients with schizophrenia treated with clozapine often gain weight. This study evaluated the effects of dietary control and physical activity among obese inpatients with schizophrenia being treated with clozapine. METHODS: Fifty three clozapine-treated obese patients with schizophrenia in a veterans hospital in eastern Taiwan who had a body mass index greater than 27 (weight divided by height in meters squared) and who were taking clozapine were randomly assigned to a study group of 28 or a control group of 25. The study group was placed on a diet that reduced calorie intake by 200 to 300 kcal per day (to 1,300 to 1,500 kcal per day for women and to 1,600 to 1,800 kcal per day for men) and a six month regimen of regular physical activity in which participants used approximately 600 to 750 kcal per week (level walking and walking on stairs for 60 minutes three days per week). Anthropometric, metabolic, and hormonal parameters were measured after three and six months by using anthropometry, an enzyme autoanalyzer, immunoassay, and enzyme-linked immunosorbent assay. RESULTS: Compared with the control group, the study group showed a significant decrease in body weight, body mass index (5.4% reduction), waist circumference (3.3 cm), and hip circumference (3.3 cm) after three months and after six months. Triglyceride and insulin-like growth factor-binding protein-3 (IGFBP-3) decreased significantly only after six months. CONCLUSIONS: A program of dietary control and regular physical activity can significantly reduce body weight and improve metabolic profiles of insulin, triglyceride, and IGFBP-3 among obese inpatients taking clozapine for the treatment of schizophrenia. PMID- 17412859 TI - Base rates of firearm possession by hospitalized psychiatric patients. AB - OBJECTIVE: The authors assessed base rates of firearm possession reported by hospitalized psychiatric patients. METHODS: The study involved retrospective review of the charts of 100 patients consecutively admitted after a hospital had incorporated routine firearm screening in the workup done at admission and the charts of 100 patients who had been consecutively admitted during an earlier interval when firearm screening had been done on an as-needed basis. RESULTS: Whereas one patient (1%) acknowledged having access to a firearm when screening had been done on an as-needed basis, under conditions of routine screening, nine patients (9%) reported owning or having access to a firearm. CONCLUSIONS: A substantial proportion of hospitalized patients have access to firearms. Unless patients are directly asked about firearm access, this information may not come to the attention of health care providers. Strategies to manage the risk of suicide and violence by hospitalized patients may benefit from routine assessment of firearms possession. PMID- 17412860 TI - Trends in the use of psychotropic drugs in Taiwan: a population-based national health insurance study, 1997-2004. AB - OBJECTIVE: This study examined changes in the use of psychotropic agents in Taiwan between 1997 and 2004. METHODS: The population-based National Health Insurance database was used to detect trends in the use of psychotropic drugs in Taiwan. A dynamic random sample of 200,432 persons was gathered and followed from 1997 to 2004. Persons in the sample who had made at least one service claim during these years for either ambulatory or inpatient care with a psychotropic drug prescription were identified. RESULTS: The prevalence of any psychotropic drug use increased from 7.1% in 1997 to 12.0% in 2004. The changes in prevalence of use of four kinds of psychotropic drugs from 1997 to 2004 were as follows: antipsychotics, 3.4% to 3.7%, although this finding was not significant; antidepressants, 2.2% to 4.4% (p<.001); mood stabilizers, .6% to 1.3% (p<.001); and anxiolytic-hypnotic drugs, 3.0% to 7.3% (p<.001). CONCLUSIONS: Overall, the prevalence of use of three kinds of psychotropic drugs (antidepressants, mood stabilizers, and anxiolytic-hypnotic drugs) increased greatly from 1997 to 2004. PMID- 17412861 TI - Inmate suicide and time spent in special disciplinary housing in New York State prison. AB - OBJECTIVE: This study examined the number of days that prisoners spent in a special disciplinary housing unit in New York State prison before a suicide occurred. METHODS: Suicides that occurred between 1993 and 2003 were reviewed. RESULTS: A total of 32 suicides occurred in one-person special housing cells during the 11-year period. Inmates spent a median of 63 days in a special disciplinary housing cell before committing suicide. The aggregate median sentence these inmates were serving in special housing at the time of suicide was 298.5 days. CONCLUSIONS: Because most suicides in a special housing unit occurred within eight weeks of placement, enhanced observation of special housing inmates is warranted in that period at a minimum. PMID- 17412862 TI - Predicting hospitalization versus discharge of suicidal patients presenting to a psychiatric emergency service. AB - OBJECTIVE: Suicidal ideation frequently prompts visits to psychiatric emergency departments, and more information is needed about factors that mediate clinicians' decisions to hospitalize or discharge patients with suicidal ideation. METHODS: The authors reviewed records for 257 patients presenting with suicidal ideation to a psychiatric emergency service. Demographic and clinical correlates of hospitalization were examined by backward stepwise binary logistic regression. RESULTS: Hospitalization occurred for 70% of suicidal persons and was significantly associated with psychosis, a history of attempted suicide, and a suicidal plan. With potential confounding factors controlled, these variables correctly classified 80% of hospitalization decisions. CONCLUSIONS: Psychosis, past suicide attempts, and the presence of a suicide plan robustly predicted the decision to hospitalize suicidal persons seen in psychiatric emergency services. Diagnosis, pharmacotherapy, having a psychiatrist, and insurance subtype were unrelated to hospitalization decisions, suggesting that psychiatric emergency department staff perceive few alternatives to hospitalization when psychosis and suicide plans accompany suicidal ideation. PMID- 17412863 TI - Use of medical emergency departments by veterans with schizophrenia. PMID- 17412864 TI - Reading disability goes beyond consent forms. PMID- 17412865 TI - Adherence to treatment in poorer countries: a new research direction? PMID- 17412866 TI - Altered gastric corpus expression of tissue inhibitors of metalloproteinases in human and murine Helicobacter infection. AB - BACKGROUND: Matrix metalloproteinases (MMPs) have roles in inflammation and other processes relevant to the architectural disturbances seen in the gastric mucosa in response to Helicobacter pylori infection. Upregulation of MMPs has been reported in H pylori infection, but there are no detailed reports regarding altered production of their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). AIMS: To investigate changes in the abundance of TIMPs in human gastric corpus mucosa and murine stomach in Helicobacter infection, and to study cellular sources in man. METHODS: Gastric corpus biopsy samples were assessed for abundance of mRNA or protein for TIMP-1 to -4 by real time quantitative PCR or western blotting, respectively. Antral and corpus biopsies were processed for histology, H pylori status and inflammatory scoring. Cellular sources of TIMP-1, -3 and -4 were examined by indirect immunohistochemistry. Circulating gastrin was measured by radioimmunoassay. Also, abundance of TIMP-1, -3 and -4 mRNA in the stomach of Helicobacter felis infected mice post-infection was compared with that of uninfected control animals. RESULTS: Compared with uninfected patients, mRNA and protein for TIMP-1, -3 and 4 were significantly more abundant in the gastric corpus of H pylori infected subjects. Gastric TIMP expression did not differ significantly between hyper- and normogastrinaemic subjects within the H pylori negative and positive groups. There was no difference in mRNA abundance for MMP-3 or -8. Immunohistochemistry showed TIMP proteins localised to gastric epithelial, stromal cells and inflammatory cells. Murine H felis infection was associated with upregulation of TIMP-1 and -3 mRNA. CONCLUSIONS: Helicobacter infection is associated with upregulation of specific TIMPs (TIMP-1 and -3) in glandular epithelium and stroma. It is suggested that increased expression of specific protease inhibitors in the corpus mucosa may exert important effects on extracellular matrix remodelling and influence the outcome of H pylori infection. PMID- 17412867 TI - Upregulation of neutrophil gelatinase-associated lipocalin in oesophageal squamous cell carcinoma: significant correlation with cell differentiation and tumour invasion. AB - BACKGROUND: Neutrophil gelatinase-associated lipocalin (NGAL) is a member of the lipocalin family. Recently, an elevated NGAL expression was reported in several types of cancers. However, the characteristics of NGAL expression in oesophageal squamous cell carcinoma (ESCC) are still unknown. AIM: To demonstrate the role of NGAL in ESCC. METHODS: NGAL expression in 81 paraffin sections, including ESCC, normal mucosa, simple hyperplasia and dysplasia, and in 73 fresh specimens of ESCC was analysed by immunohistochemistry, western blot and gelatin zymography. RESULTS: On immunohistochemical study, ESCC showed a diverse staining pattern for NGAL. However, only a weak positive signal was present within a restricted cytoplasmic area in the normal oesophageal epithelium. In dysplasia, altered NGAL expression could also be observed. On western blot study, NGAL expression level was found to be significantly higher in ESCC than in normal mucosa (p=0.030), and to be positively correlated with cell differentiation. However, no significant association was observed between NGAL expression and cell proliferation. In addition, the enzymic activity of the NGAL/matrix metalloproteinase 9 complex was much higher in ESCC than in normal mucosa, and was significantly correlated with the depth of tumour invasion in zymography analysis (p=0.006). CONCLUSIONS: The findings suggest that NGAL is involved in the differentiation pathway and invasive progression of ESCC. PMID- 17412868 TI - Alteration in human defensin-5 expression following gastric bypass surgery. AB - BACKGROUND: Roux-en-Y gastric bypass surgery provides a novel human model to investigate small bowel mucosal innate immunity, in which there is loss of gastric acid-mediated protection against orally-acquired microorganisms. AIM: To study changes in jejunal mucosal immunoreactivity of human defensin (HD)-5, an antimicrobial peptide normally produced by Paneth cells. METHODS: Mucosal samples were obtained from 18 female patients (24-54 years), from the same segment of jejunum during and after gastric bypass surgery. Samples were used for bacterial culture and immunohistochemistry using anti-HD-5 antibody. The number of immunoreactive cells per crypt and villus were determined and expressed as mean (SD). RESULTS: No bacteria were cultured from any of the perioperative jejunal samples but colonies of bacteria normally present in the pharynx were identified during culture of all postoperative jejunal biopsy specimens (1->100 colonies). Paneth cell numbers per crypt were unchanged after gastric bypass (4.16 (0.71) vs 4.24 (0.78)). However, following surgery, there was an increase in HD-5-positive intermediate cells per crypt (0.25 (0.41) vs 1.12 (0.66), p<0.01), HD-5 staining enterocytes per crypt (0.03 (0.09) vs 1.38 (1.10), p<0.01), HD-5 staining material in the crypt lumen (crypt lumens: 5.0% (10.9%) vs 68.1% (27.9%), p<0.01) and HD-5 immunoreactivity coating the luminal surface of villus enterocytes (villi sampled: 15.0% (31.0%) vs 67.5% (42.0%), p<0.01). CONCLUSIONS: Bacteria normally resident in the pharynx were present in the proximal jejunal mucosa following Roux-en-Y gastric bypass surgery. After gastric bypass, there was increased secretion of HD-5 and an increase in HD-5 expressing intermediate cells and enterocytes in the crypt. The increase in HD-5 expression in the jejunal mucosa following gastric bypass surgery is likely to be secondary to exposure to orally-acquired microorganisms. PMID- 17412869 TI - Comparative proteomic analysis for the detection of biomarkers in pancreatic ductal adenocarcinomas. AB - AIMS: To search for novel potential protein biomarkers for the early detection and better intervention of pancreatic ductal adenocarcinoma (PDAC). METHODS: Eight pairs of matched PDAC and non-cancerous pancreas tissues were profiled with two-dimensional electrophoresis; differentially expressed proteins were identified by mass spectrometry. Expression patterns of TBX4 (T-box transcription factor TBX4) and HSP60 (60 KDa heat shock protein) were studied with immunohistochemistry using tissue microarrays. RESULTS: A total of 48 differentially expressed proteins were identified; 30 of them are novel potential biomarkers. Immunohistochemistry showed that TBX4 expression could be seen in both centroacinar cells and small ducts in normal pancreas and tumour cells in 5/5 (100%) well differentiated, 35/38 (92.1%) moderately differentiated, and 11/18 (61.1%) poorly differentiated PDAC tissues with different staining intensity. However, in normal acinar cells and tumour cells in the other 3/38 (7.9%) moderately differentiated and 7/18 (38.9%) poorly differentiated PDAC tissues, there was no visible TBX4 expression. The expression difference of TBX4 between moderately differentiated and poorly differentiated PDAC tissues was statistically significant (p<0.01). In addition, there was obvious morphology difference between TBX4 negatively stained and positively stained tumour cells, which suggests different cellular origins. Strong expression of HSP60 could be seen in both acinar cells and small ducts in normal pancreas tissues and tumour cells in PDAC tissues except for islets and tumour stoma; no correlation was found between HSP60 expression and differentiation of PDAC tissues. CONCLUSIONS: 30 novel potential biomarkers differentially expressed in PDAC tissues were identified. TBX4 may be a differentiation related protein; its prognostic value for PDAC deserves further study. PMID- 17412870 TI - Central HER2 IHC and FISH analysis in a trastuzumab (Herceptin) phase II monotherapy study: assessment of test sensitivity and impact of chromosome 17 polysomy. AB - AIMS: To investigate the correlation between centrally assessed human epidermal growth factor receptor 2 (HER2) immunohistochemistry (IHC) and fluorescence in situ hybridisation (FISH) results and response to treatment of patients with metastatic breast cancer enrolled in a first-line, phase II, open-label, 3-weekly trastuzumab (Herceptin) monotherapy trial (WO16229). METHODS: Samples from participants in the WO16229 trial were collected and tumour HER2 status determined by IHC and FISH. HER2 test results were interpreted according to manufacturers' test kit protocols. Responders were defined as patients showing either partial or complete responses. RESULTS: Response data were available for 103/105 patients; centrally confirmed HER2 status was available for 95 patients. Intra-laboratory concordance for central IHC and FISH results was 93%. Complete responses were seen in two patients; their samples were IHC 3+ and FISH positive. Partial responses were seen in 17 patients; all were IHC 3+ and 14 were FISH positive. IHC and FISH showed 100% and 84.2% sensitivity, respectively, in determining response to trastuzumab. Polysomy was observed in 27% of patients; six responded to trastuzumab treatment. All six responders showed HER2 overexpression (IHC 3+) and HER2 gene amplification; two were FISH negative due to chromosome 17 polysomy. CONCLUSIONS: HER2 determination by IHC and FISH correlates with clinical response data in the WO16229 trial with high concordance of IHC and FISH results. Polysomy is the major cause of response in FISH-negative cases; polysomic cases should be retested by strictly standardised IHC. PMID- 17412871 TI - Persistent parvovirus b19 infection resulting in carpal tunnel syndrome. PMID- 17412872 TI - Comparing subjective and digital image analysis HER2/neu expression scores with conventional and modified FISH scores in breast cancer. AB - BACKGROUND: HER2/neu expression and fluorescence in situ hybridisation (FISH) amplification have therapeutic significance. AIMS: To compare subjective HER2/neu expression scores with digital image analysis (DIA) and conventional and modified FISH scores in breast cancer. METHODS: Sixty HercepTest-immunostained breast carcinomas, prospectively scored as consensus 2+ and 3+ (DAKO protocol) by two observers, were analysed with DIA, and conventional (Vysis) and modified FISH scoring protocols. RESULTS: With consensus scoring, 23 (38%) of the 60 cases were 2+ and 37 (62%) were 3+. Agreement with DIA scores was 100%. With conventional FISH scoring, 4 of the 3+ cases did not show amplification, but all of those negative cases had high HER2/neu copy numbers. With the modified FISH scoring protocol, all HercepTest immunohistochemical 3+ cases were amplified. Of the 2+ cases, 3 were amplified with the modified FISH protocol and 4 with the conventional FISH protocol. CONCLUSIONS: Modified FISH scores were better correlated with HercepTest 3+ consensus and DIA scores than were conventional FISH scores. HER2/neu DIA scoring is a cost-effective supplementary tool in surgical pathology. PMID- 17412873 TI - Antigenicity testing by immunohistochemistry after tissue oxidation. AB - AIMS: Archived tissue blocks preserve the antigenicity of samples for a long time under normal storage conditions, whereas tissue sections may show a diminished immunoreactivity over time. Little is known about the processes responsible for antigenicity loss and how tissue sections should be conserved for extended storage. Oxidation and drying are presumed mechanisms of antigenicity loss. To prove this, degradation of immunoreactivity was provoked by chemical oxidation, photo-oxidation and artificial drying. METHODS: First, paraffin sections of an oestrogen receptor (ER) positive breast carcinoma were subjected for variable time periods to H2O2, ultraviolet A (UVA) irradiation and dry heat (56 degrees C) prior to ER immunohistochemistry. Second, using heat and UVA irradiation several other antigens (ER, PR, HER-2neu, p53, p63, p16, PSA, CK5/6, CK7, CK20, SMA, Fli 1, c-kit, CD20 and EGFR) were tested, using internal control tissue microarray sections. RESULTS: While H2O2 had no effect on the ER-staining intensity, extended drying showed a detectable decrease in staining after 10 days, and UVA irradiation induced a decrease after 3 days. Entire antigenicity loss was not observed. Except for HER-2neu, PSA and Fli-1, all antigens showed some diminution in antigenicity, but no entire antigenicity loss, after heating and UVA irradiation. CONCLUSIONS: This study confirms that photo-oxidation and drying have an influence on immunohistochemistry outcome, and protocols for testing the stability of specific antigens are provided. PMID- 17412874 TI - Transfusion in sub-Saharan Africa: does a Western model fit? AB - This review examines the current state of transfusion services in sub-Saharan Africa and presents the argument for and against the Western model of a centralised blood service with 100% voluntary non-remunerated blood donors as advocated by the World Health Organization. The current practice of family replacement donors in hospital-based blood service is the most economical option, but in the face of high child and maternal mortality rates the blood supply has proved to be insufficient. With estimates of 5-10% of HIV transmission in Africa being the result of contaminated blood transfusions, there is a need to improve testing for transfusion transmissible diseases and the selection of blood donors. Of major concern, with respect to testing, is the quality of kits being used and the continuity of supply. The need to produce components is discussed in the context of the transfusion needs in sub-Saharan Africa. The running costs of establishing and maintaining centralised blood services need careful consideration as such projects need to be sustainable in the future. It is concluded that both options are viable while centralised programmes are being developed, and a pragmatic approach should be taken to ensure that the patients' needs are met and that resources are suitably utilised to ensure sustainability. PMID- 17412875 TI - Laboratory costs of a hospital-based blood transfusion service in Malawi. AB - BACKGROUND: Despite policies advocating centralised transfusion services based on voluntary donors, the hospital-based replacement donor system is widespread in sub-Saharan Africa. AIMS: To evaluate the cost of all laboratory resources needed to provide a unit of safe blood in rural Malawi using the family replacement donor system METHODS: Full economic costs of all laboratory tests used to screen potential donors and to perform cross-matching were documented in a prospective, observational study in Ntcheu district hospital laboratory. RESULTS: 1729 potential donors were screened and 11,008 tests were performed to ensure that 1104 units of safe blood were available for transfusion. The annual cost of all transfusion-related tests (in 2005 USdollars) was USdollars 17,976, equivalent to USdollars 16.28 per unit of transfusion-ready blood. Transfusion-related tests used 53% of the laboratory's total annual expenditure of USdollars 33,608. CONCLUSIONS: This is the first study to provide prospective economic costs of all laboratory tests associated with the family replacement donor system in a district hospital in Africa. Results show that despite potential economies of scale, a unit of blood from the centralised system costs about three times as much as one from the hospital-based "replacement" system. Factors affecting these relative costs are complex but are in part due to the cost of donor recruitment in centralised systems. In the replacement system the cost of donor recruitment is entirely borne by families of patients needing a blood transfusion. PMID- 17412876 TI - Unrepresentative astrocytoma biopsy sampling is partly overcome by assessment of the MIB-1-labelled growth fraction. PMID- 17412877 TI - Attenuated aerobic exercise capacity in CD36 deficiency. AB - BACKGROUND: An important role of CD36 in muscle fatty acid (FA) uptake has been shown in CD36-knockout or CD36-overexpressed mice. FA is a predominant substrate in energy production during light exercise below the anaerobic threshold (AT). We studied whether aerobic exercise capacity in humans could be affected by CD36 deficiency. METHODS: We investigated the ventilatory threshold (VT) and serum FA changes in normal participants (n = 22) and participants with CD36 deficiency (n = 12) during pedalling on a cycle ergometer. RESULTS: In participants with CD36 deficiency, FA levels were not reduced at peak work rate, whereas FA levels decreased by about 50% in normal participants. Participants with CD36 deficiency showed significantly lower VT than normal participants. A significant correlation was observed between VT and percentage changes in FA at peak work rate. CONCLUSION: This study found reduced FA utilisation and an attenuated aerobic exercise capacity in CD36 deficiency, indicating that CD36-mediated FA oxidation is an important determinant for aerobic exercise capacity in humans. PMID- 17412879 TI - Myopathy caused by HRAS germline mutations: implications for disturbed myogenic differentiation in the presence of constitutive HRas activation. AB - BACKGROUND: Rare reports on patients with congenital myopathy with excess of muscle spindles (CMEMS), hypertrophic cardiomyopathy and variable features resembling Noonan syndrome have been published, but the genetic basis of this condition is so far unknown. METHODS AND RESULTS: We analysed PTPN11 and RAS genes in five unrelated patients with this phenotype, and found HRAS mutations in four of them. Two disease-associated mutations, G12V and G12S, have previously been observed in patients with Costello syndrome (CS), and two other mutations, E63K and Q22K, are novel. All four mutations are predicted to enhance downstream HRas signalling, suggesting that CMEMS is a developmental consequence of sustained HRas activation in skeletal muscle. CONCLUSION: This type of myopathy may represent a previously unrecognized manifestation of CS. However, some patients carrying HRAS mutations may exhibit prominent congenital muscular dysfunction, although features of CS may be less obvious, suggesting that germline HRAS mutations may underlie some cases of otherwise unclassified neonatal neuromuscular disorders. PMID- 17412880 TI - Partial deletions are associated with an increased risk of complete deletion in AZFc: a new insight into the role of partial AZFc deletions in male infertility. AB - BACKGROUND: The AZFc region on the human Y chromosome has been found to be functionally important in spermatogenesis. Complete AZFc deletion is one of the most frequent causes of male infertility and the roles of partial AZFc deletions (gr/gr and b2/b3 deletions) in spermatogenesis are controversial. METHODS: To further study the roles of partial AZFc deletions in spermatogenic impairment and the relationship between complete and partial AZFc deletions, these deletions were typed and quantitative analysis of DAZ gene copies and Y chromosome haplogrouping were performed for seven pedigrees of complete AZFc deletion carriers, comprising 296 infertile and 280 healthy Chinese men. RESULTS: Neither the gr/gr nor the b2/b3 deletion was found to be associated with spermatogenic failure. In one pedigree, a complete AZFc deletion was observed to result from the gr/gr deletion, suggesting that complete deletions of AZFc can be preceded by partial deletions. In addition, a new gr/gr-deleted Y haplogroup Q1 was identified and the reported fixation of the b2/b3 deletion in haplogroup N confirmed. The frequency of complete AZFc deletion in haplogroups Q1 and N was significantly higher than that in the other haplogroupsm with fewer partial deletions. Duplications of DAZ gene copies were also observed in this study. CONCLUSIONS: To date, these observations comprise the first evidence showing that partial AZFc deletions can increase the risk of complete AZFc deletion. The susceptibility of partial AZFc deletions to complete AZFc deletion deserves further examination, especially in the populations or Y haplogroups abundant in partial AZFc deletions. PMID- 17412882 TI - Connexin 50 gene on human chromosome 1q21 is associated with schizophrenia in matched case control and family-based studies. AB - BACKGROUND: The gap junction subunit connexin permits direct intercellular exchange of ions and molecules including glutamate, and plays an important role in the central nervous system. The connexin 40 (Cx40) and connexin 50 (Cx50) genes are located on chromosome 1q21.1, a region strongly linked with schizophrenia. These lines of evidence suggest that Cx40 and Cx50 may play a role in schizophrenia. METHODS: Using an allele-specific PCR assay, four polymorphisms each were genotyped for Cx40 and Cx50 in 190 Caucasian patients with schizophrenia and 190 controls matched for sex, age and ethnicity. Following up, Cx50 rs989192 and rs4950495 were investigated in 99 Canadian and 163 Portuguese trios and nuclear families with schizophrenia probands. Hardy-Weinberg equilibrium and linkage disequilibrium (LD) block identification was carried out with HaploView, and association analysis for alleles and haplotypes with a permutation test of 10 000 simulations was carried out using the UNPHASED software program. RESULTS: Distributions of genotype frequencies of all markers were in Hardy-Weinberg equilibrium in Caucasian patients, controls and families. One rs989192-rs4950495 LD block was found in patients but not in controls. We found a significant association between the Cx50 rs989192-rs4950495 haplotype and schizophreniay (chi(2) = 29.55, p<0.01). The A-C haplotype had a higher frequency in patients (chi(2) = 7.153, p<0.01). Family studies also showed that the A-C haplotype was transmitted more often to patients with schizophrenia (chi(2) = 8.43, p<0.01). No association of Cx40 with schizophrenia was found for allele, genotype or haplotype analyses. CONCLUSIONS: Our matched case-control and family study indicate that Cx50, but not Cx40, may play a role in the genetic susceptibility to schizophrenia. PMID- 17412883 TI - The fatty acid amide hydrolase inhibitor URB597 (cyclohexylcarbamic acid 3' carbamoylbiphenyl-3-yl ester) reduces neuropathic pain after oral administration in mice. AB - Fatty acid amide hydrolase (FAAH) is an intracellular serine hydrolase that catalyzes the cleavage of bioactive fatty acid ethanolamides, such as the endogenous cannabinoid agonist anandamide. Genetic deletion of the faah gene in mice elevates brain anandamide levels and amplifies the antinociceptive effects of this compound. Likewise, pharmacological blockade of FAAH activity reduces nocifensive behavior in animal models of acute and inflammatory pain. In the present study, we investigated the effects of the selective FAAH inhibitor URB597 (KDS-4103, cyclohexylcarbamic acid 3'-carbamoylbiphenyl-3-yl ester) in the mouse chronic constriction injury (CCI) model of neuropathic pain. Oral administration of URB597 (1-50 mg/kg, once daily) for 4 days produced a dose-dependent reduction in nocifensive responses to thermal and mechanical stimuli, which was prevented by a single i.p. administration of the cannabinoid CB(1) receptor antagonist rimonabant (1 mg/kg). The antihyperalgesic effects of URB597 were accompanied by a reduction in plasma extravasation induced by CCI, which was prevented by rimonabant (1 mg/kg i.p.) and attenuated by the CB(2) antagonist SR144528 (1 mg/kg i.p.). Oral dosing with URB597 achieved significant, albeit transient, drug levels in plasma, inhibited brain FAAH activity, and elevated spinal cord anandamide content. The results provide new evidence for a role of the endocannabinoid system in pain modulation and reinforce the proposed role of FAAH as a target for analgesic drug development. PMID- 17412884 TI - Apomine enhances the antitumor effects of lovastatin on myeloma cells by down regulating 3-hydroxy-3-methylglutaryl-coenzyme A reductase. AB - Apomine, a 1,1-bisphosphonate-ester with antitumor activity, has previously been reported to strongly down-regulate 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CoA reductase), the rate-limiting enzyme in the mevalonate pathway responsible for the prenylation of proteins. Here, we show that although apomine down-regulated HMG-CoA reductase protein levels in myeloma cells, it did not inhibit protein prenylation, and apomine-induced apoptosis could not be prevented by mevalonate, indicating that apomine cytotoxicity is independent from its effects on HMG-CoA reductase. Instead, apomine cytotoxicity was prevented by the addition of phosphatidylcholine, which is similar to the previously reported ability of phosphatidylcholine to overcome the cytotoxicity of farnesol, whereas phosphatidylcholine had no effect on down-regulation of HMG-CoA reductase by apomine. These findings raised the possibility that apomine, independent from its own cytotoxic effects, could enhance the antitumor effects of the competitive HMG CoA reductase inhibitor lovastatin via down-regulating HMG-CoA reductase. Indeed, treatment with apomine in combination with lovastatin resulted in synergistic decreases in viable cell number and induction of apoptosis. At the concentrations used, apomine down-regulated HMG-CoA reductase protein levels without being cytotoxic. Accumulation of unprenylated Rap1A by lovastatin was enhanced in the presence of apomine. Furthermore, synergy was completely prevented by mevalonate, and apomine did not synergize with desoxolovastatin, which does not inhibit HMG CoA reductase. We conclude that the synergistic drug interaction results from an enhancement by apomine of the effects of lovastatin, mediated by down-regulation of HMG-CoA reductase by apomine. Thus, these findings demonstrate a novel strategy for enhancing the antitumor effects of lovastatin. PMID- 17412885 TI - CD4+CD25high Foxp3+ regulatory T cells in myelodysplastic syndrome (MDS). AB - Foxp3+ regulatory T cells (Tregs) play a central role in maintaining immune tolerance. A reduction in the function of Tregs is a key feature of autoimmune diseases, whereas their expansion in malignant diseases leads to the suppression of host antitumor responses. We analyzed the absolute number of CD4+ and CD8(+) Tregs in the peripheral blood of 52 patients with myelodysplastic syndrome (MDS) and show a significant correlation between increased number of CD4+ Tregs and MDS subgroups with 5% or more bone marrow blasts (P < .001), high International Prognostic Scoring System (IPSS) score (P < .001), and disease progression (P < .001), whereas no correlation between CD8+ Tregs and prognostic variables was observed. The CD4+ Tregs showed a polyclonal spectratype, and the percentage of the naive subset was significantly higher in the high-risk patients compared with low-risk or healthy age-matched donors (P = .032). Our data suggest that CD4+ Treg expansion is a feature of high-risk MDS and progression to aggressive subtypes of the disease. PMID- 17412886 TI - High PR3 or ELA2 expression by CD34+ cells in advanced-phase chronic myeloid leukemia is associated with improved outcome following allogeneic stem cell transplantation and may improve PR1 peptide-driven graft-versus-leukemia effects. AB - The primary granule proteins elastase (ELA2) and proteinase 3 (PR3) both contain the nonapeptide PR1, which can induce cytotoxic T lymphocyte (CTL) responses against chronic myeloid leukemia (CML) cells. To investigate whether eradication of CML after allogeneic stem cell transplantation (SCT) was influenced by PR3 and ELA2 gene expression or PR1-specific CTL responses, we studied cells from 87 CML patients and 27 HLA-A*0201(+) donors collected prior to T-cell-depleted HLA identical sibling SCT. For patients in advanced phase (AdP), a higher expression of both PR3 and ELA2 in CD34(+) progenitors before SCT was associated with a lower incidence of relapse-related death, improved leukemia-free survival (LFS), and overall survival (OS); in chronic phase patients, no differences were observed. PR1-CTL responses were detected in 7 of 27 HLA-identical sibling donors, and associated with improved LFS and OS after SCT on follow-up. PR1-CTL responses detected in 7 of 28 CML patients before transplantation were not predictive of outcome and correlated inversely with PR3 and ELA2 expression. These findings suggest that assessment of PR3 and ELA2 expression in leukemic progenitors is useful for predicting posttransplantation outcome in AdP patients undergoing SCT. The presence of a donor immune response against PR1 may be advantageous and could be exploited therapeutically. PMID- 17412887 TI - Acute myeloid leukemia with the 8q22;21q22 translocation: secondary mutational events and alternative t(8;21) transcripts. AB - Nonrandom and somatically acquired chromosomal translocations can be identified in nearly 50% of human acute myeloid leukemias. One common chromosomal translocation in this disease is the 8q22;21q22 translocation. It involves the AML1 (RUNX1) gene on chromosome 21 and the ETO (MTG8, RUNX1T1) gene on chromosome 8 generating the AML1-ETO fusion proteins. In this review, we survey recent advances made involving secondary mutational events and alternative t(8;21) transcripts in relation to understanding AML1-ETO leukemogenesis. PMID- 17412888 TI - Endothelial potential of human embryonic stem cells. AB - Growing interest in using endothelial cells for therapeutic purposes has led to exploring human embryonic stem cells as a potential source for endothelial progenitor cells. Embryonic stem cells are advantageous when compared with other endothelial cell origins, due to their high proliferation capability, pluripotency, and low immunogenity. However, there are many challenges and obstacles to overcome before the vision of using embryonic endothelial progenitor cells in the clinic can be realized. Among these obstacles is the development of a productive method of isolating endothelial cells from human embryonic stem cells and elucidating their differentiation pathway. This review will focus on the endothelial potential of human embryonic stem cells that is described in current studies, with respect to the differentiation of human embryonic stem cells to endothelial cells, their isolation, and their characterization. PMID- 17412889 TI - PECAM-1: a multifaceted regulator of megakaryocytopoiesis. AB - PECAM-1 (CD31) knockout (KO) mice exhibit excessive megakaryocytopoiesis accompanied by increased numbers of megakaryocytes associated with the stromal niche rather than the vascular niche. During earlier stages of megakaryocytopoiesis in KO marrow, an expanded Lin(-)Sca-1(+) c-kit(+) hematopoietic stem cell (HSC) population and increased quiescent Lin(-) progenitor pool were identified. During the later stages of megakaryocytopoiesis, CD31KO megakaryocytes exhibited abnormal adhesion/transmigration behaviors. Lastly, KO animals exhibited excessive splenic extramedullary megakaryocytopoiesis, which likely compensates for the impaired marrow megakaryocytopoiesis, resulting in normal peripheral platelet number. Thus, PECAM 1 modulates megakaryocytopoiesis in a hierarchic manner, functioning as a thermostat to "fine-tune" megakaryocytopoiesis. PMID- 17412890 TI - Increased adhesion to endothelial cells of erythrocytes from patients with polycythemia vera is mediated by laminin alpha5 chain and Lu/BCAM. AB - Patients with polycythemia vera (PV) have a JAK2 (a cytosolic tyrosine kinase) mutation and an increased risk of vascular thrombosis related to red blood cell (RBC) mass and platelet activation. We investigated functional RBC abnormalities that could be involved in thrombosis. RBC adhesion to human umbilical vein endothelial cells (HUVECs) was measured by a radiometric technique and in a flow system by video microscopy, and adhesion molecule expression was determined using specific antibodies (against CD36, CD49d, ICAM-4, Lu/BCAM, CD147, and CD47) and flow cytometry in a group of 38 patients with PV and a group of 36 healthy volunteers. Adhesion of PV RBCs was 3.7-fold higher than that of normal RBCs (P < .001). Adhesion was inhibited when PV RBCs were incubated with anti-Lutheran blood group/basal cell adhesion molecule (Lu/BCAM) or when HUVECs were treated with anti-laminin alpha(5) and to a lesser extent with anti-alpha(3) integrin. Lu/BCAM was constitutively phosphorylated in PV RBCs. Transfection of K562 cells with JAK2 617V>F resulted in increased expression and phosphorylation of Lu/BCAM. Phosphorylation of Lu/BCAM increases RBC adhesion. Our results indicate that JAK2 mutation might be linked to Lu/BCAM modification and increased RBC adhesiveness, which may be a factor favoring thrombosis in PV. PMID- 17412891 TI - Lineage specification of hematopoietic stem cells: mathematical modeling and biological implications. AB - Lineage specification of hematopoietic stem cells is considered a progressive restriction in lineage potential. This view is consistent with observations that differentiation and lineage specification is preceded by a low-level coexpression of lineage specific, potentially antagonistic genes in early progenitor cells. This coexistence, commonly referred to as priming, disappears in the course of differentiation when certain lineage-restricted genes are upregulated while others are downregulated. Based on this phenomenological description, we propose a quantitative model that describes lineage specification as a competition process between different interacting lineage propensities. The competition is governed by environmental stimuli promoting a drift from a multipotent coexpression to the dominance of one lineage. The assumption of a context dependent intracellular differentiation control is consistently embedded into our previously proposed model of hematopoietic stem cell organization. The extended model, which comprises self-renewal and lineage specification, is verified using available data on the lineage specification potential of primary hematopoietic stem cells and on the differentiation kinetics of the FDCP-mix cell line. The model provides a number of experimentally testable predictions. From our results, we conclude that lineage specification is best described as a flexible and temporally extended process in which lineage commitment emerges as the result of a sequence of small decision steps. The proposed model provides a novel systems biological view on the functioning of lineage specification in adult tissue stem cells and its connections to the self-renewal properties of these cells. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412892 TI - A modified polymerase chain reaction-long serial analysis of gene expression protocol identifies novel transcripts in human CD34+ bone marrow cells. AB - Transcriptome profiling offers a powerful approach to investigating developmental processes. Long serial analysis of gene expression (LongSAGE) is particularly attractive for this purpose because of its inherent quantitative features and independence of both hybridization variables and prior knowledge of transcript identity. Here, we describe the validation and initial application of a modified protocol for amplifying cDNA preparations from <10 ng of RNA (<10(3) cells) to allow representative LongSAGE libraries to be constructed from rare stem cell enriched populations. Quantitative real-time polymerase chain reaction (Q-RT-PCR) analyses and comparison of tag frequencies in replicate LongSAGE libraries produced from amplified and nonamplified cDNA preparations demonstrated preservation of the relative levels of different transcripts originally present at widely varying levels. This PCR-LongSAGE protocol was then used to obtain a 200,000-tag library from the CD34+ subset of normal adult human bone marrow cells. Analysis of this library revealed many anticipated transcripts, as well as transcripts not previously known to be present in CD34+ hematopoietic cells. The latter included numerous novel tags that mapped to unique and conserved sites in the human genome but not previously identified as transcribed elements in human cells. Q-RT-PCR was used to demonstrate that 10 of these novel tags were expressed in cDNA pools and present in extracts of other sources of normal human CD34+ hematopoietic cells. These findings illustrate the power of LongSAGE to identify new transcripts in stem cell-enriched populations and indicate the potential of this approach to be extended to other sources of rare cells. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412893 TI - Isolation, characterization, and differentiation to hepatocyte-like cells of nonparenchymal epithelial cells from adult human liver. AB - Activation and proliferation of human liver progenitor cells has been observed during acute and chronic liver diseases. Our goal was to investigate the presence of these putative progenitors in the liver of patients who underwent lobectomy for various reasons but did not show any hepatic insufficiency. Hepatic lesions were evaluated by histological analysis. Nonparenchymal epithelial (NPE) cells were isolated from samples of human liver resections located at a distance from the lesion that motivated the operation and were cultured and characterized. These cells exhibited a marked proliferative potential. They did not express the classic set of stem cell/progenitor markers (Oct-4, Rex-1, alpha-fetoprotein, CD90, c-kit, and CD34) and were faintly positive for albumin. When cultured at confluence in the presence of hepatocyte growth factor and either epidermal growth factor or fibroblast growth factor-4, they entered a differentiation process toward hepatocytes. Their phenotype was quantitatively compared with that of mature human hepatocytes in primary culture. Differentiated NPE cells expressed albumin; alpha1-antitrypsin; fibrinogen; hepatobiliary markers such as cytokeratins 7, 19, and 8/18; liver-enriched transcription factors; and genes characterized by either a fetal (cytochrome P4503A7 and glutathione S-transferase pi) or a mature (tyrosine aminotransferase, tryptophan 2,3-dioxygenase, glutathione S-transferase alpha, and cytochrome P4503A4) expression pattern. NPE cells could be isolated from the liver of several patients, irrespective of the absence or presence of lesions, and differentiated toward hepatocyte-like cells with an intermediate hepatobiliary and mature/immature phenotype. These cells are likely to represent a resident progenitor population of the adult human liver, even in the absence of hepatic failure. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412894 TI - Spheres isolated from 9L gliosarcoma rat cell line possess chemoresistant and aggressive cancer stem-like cells. AB - The rat 9L gliosarcoma is a widely used syngeneic rat brain tumor model that closely simulates glioblastoma multiforme when implanted in vivo. In this study, we sought to isolate and characterize a subgroup of cancer stem-like cells (CSLCs) from the 9L gliosarcoma cell line, which may represent the tumor initiating subpopulation of cells. We demonstrate that these CSLCs form clonal derived spheres in media devoid of serum supplemented with the mitogens epidermal growth factor and basic fibroblast growth factor, express the NSC markers Nestin and Sox2, self-renew, and differentiate into neuron-like and glial cells in vitro. More importantly, these cells can propagate and recapitulate tumors when implanted into the brain of syngeneic Fisher rats, and they display a more aggressive course compared with 9L gliosarcoma cells grown in monolayer cultures devoid of mitogens. Furthermore, we compare the chemosensitivity and proliferation rate of 9L gliosarcoma cells grown as a monolayer to those of cells grown as floating spheres and show that the sphere-generated cells have a lower proliferation rate, are more chemoresistant, and express several antiapoptosis and drug-related genes, which may prove to have important clinical implications. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412895 TI - Targeted delivery of CX3CL1 to multiple lung tumors by mesenchymal stem cells. AB - MSCs are nonhematopoietic stem cells capable of differentiating into various mesoderm-type cells. MSCs have been considered to be a potential vehicle for cell based gene therapy because MSCs are relatively easily expanded in vitro and have the propensity to migrate to and proliferate in the tumor tissue after systemic administration. Here, we demonstrated the tropism of mouse MSCs to tumor cells in vitro and multiple tumor tissues in the lung after i.v. injection of green fluorescent protein-positive MSCs in vivo. We transduced CX3CL1 (fractalkine), an immunostimulatory chemokine, to the mouse MSCs ex vivo using an adenoviral vector with the Arg-Gly-Asp-4C peptide in the fiber knob. Intravenous injection of CX3CL1-expressing MSCs to the mice bearing lung metastases of C26 and B16F10 cells strongly inhibited the development of lung metastases and thus prolonged the survival of these tumor-bearing mice. This antitumor effect depended on both innate and adaptive immunity. These results suggest that MSCs can be used as a vehicle for introducing biological agents into multiple lung tumor tissues. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412896 TI - Akt is a key modulator of endothelial progenitor cell trafficking in ischemic muscle. AB - Trafficking of transplanted endothelial progenitor cells (EPCs) to ischemic tissue is enhanced by stromal-derived factor 1 (SDF-1) and vascular endothelial growth factor (VEGF). However, it has not been studied how these cytokines modulate the local milieu to entrap EPCs. This study was performed to elucidate a molecular pathway of trafficking EPCs through Akt and to test its application as an adjuvant modality to increase EPC homing. In a mouse hind limb ischemia model, systemically administered 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine labeled mouse EPCs showed three stages of homing to ischemic limb: adhesion to endothelial cells (ECs), incorporation to capillary, and transendothelial migration into extravascular space. As an underlying mechanism to control adhesion of EPCs to ECs, we found that Akt was activated in ECs of ischemic muscle by ischemia-induced VEGF and SDF-1. In vitro and in vivo experiments using adenoviral vector for constitutively active or dominant-negative Akt genes showed that activated Akt enhanced intercellular adhesion molecule 1 (ICAM-1) expression on ECs. Akt activation in ECs also enhanced EPC incorporation to ECs and transendothelial migration in vitro experiments. Activated Akt was sufficient for induction of EPC homing even in normal hind limb, where VEGF or SDF-1 was not increased. Finally, local Akt gene transfer to ischemic limb significantly enhanced homing of systemically administered EPCs, new vessel formation, blood flow recovery, and tissue healing. Akt plays a key role in EPC homing to ischemic limb by controlling ICAM-1 and transendothelial migration. Modulation of Akt in the target tissue may be an adjunctive measure to enhance homing of systemically administered stem cells, suggesting a possibility of cell-and-gene hybrid therapy. Disclosure of potential conflicts of interest is found at the end of this article. PMID- 17412897 TI - Measuring flux of soil fumigants using the aerodynamic and dynamic flux chamber methods. AB - Methods for measuring and estimating flux density of soil fumigants under field conditions are important for the purpose of providing inputs to air dispersion models and for comparing the effects of management practices on emission reduction. The objective of this study was to measure the flux of 1,3 dichloropropene (1,3-D) and chloropicrin at a site in Georgia (GA) using the aerodynamic method and the dynamic flux chamber (FC) method. A secondary objective was to compare the effects of high density polyethylene (HDPE), and virtually impermeable film (VIF) tarps on fumigant flux at a site in Florida (FL). Chloropicrin and 1,3-D were applied by surface drip application of In-Line soil fumigant on vegetable beds covered by low density polyethylene (LDPE), HDPE, or VIF. The surface drip fumigation using In-Line and LDPE tarp employed in this study resulted in volatilization of 26.5% of applied 1,3-D and 11.2% of the applied chloropicrin at the GA site, as determined using the aerodynamic method. Estimates of mass loss obtained from dynamic FCs were 23.6% for 1,3-D and 18.0% for chloropicrin at the GA site. Flux chamber trials at the FL site indicate significant additional reduction in flux density, and cumulative mass loss when VIF tarp is used. This study supports the use of dynamic FCs as a valuable tool for estimating gas flux density from agricultural soils, and evaluating best management practices for reducing fumigant emissions to the atmosphere. PMID- 17412898 TI - Bacterial reduction of selenium in coal mine tailings pond sediment. AB - Sediment from a storage facility for coal tailings solids was assessed for its capacity to reduce selenium (Se) by native bacterial community. One Se(6+) reducing bacterium Enterobacter hormaechei (Tar11) and four Se(4+)-reducing bacteria, Klebsiella pneumoniae (Tar1), Pseudomonas fluorescens (Tar3), Stenotrophomonas maltophilia (Tar6), and Enterobacter amnigenus (Tar8) were isolated from the sediment. Enterobacter hormaechei removed 96% of the added Se(6+) (0.92 mg L(-1)) from the effluents when Se(6+) was determined after 5 d of incubation. Analysis of the red precipitates showed that Se(6+) reduction resulted in the formation of spherical particles (<1.0 microm) of Se(0) as observed under scanning electron microscope (SEM) and confirmed by EDAX. Selenium speciation was performed to examine the fate of the added Se(6+) in the sediment with or without addition of Enterobacter hormaechei cells. More than 99% of the added Se(6+) (approximately 2.5 mg L(-1)) was transformed in the nonsterilized sediment (without Enterobacter hormaechei cells) as well as in the sterilized (heat-killed) sediment (with Enterobacter hormaechei cells). The results of this study suggest that the lagoon sediments at the mine site harbor Se(6+)- and Se(4+)-reducing bacteria and may be important sinks for soluble Se (Se(6+) and Se(4+)). Enterobacter hormaechei isolated from metal-contaminated sediment may have potential application in removing Se from industrial effluents. PMID- 17412899 TI - Mitigation of shallow groundwater nitrate in a poorly drained riparian area and adjacent cropland. AB - Riparian ecosystems, through their unique position in the agricultural landscape and ability to influence nutrient cycles, can potentially reduce NO3 loading to surface and ground waters. The purpose of this study was to determine the fate of NO3 in shallow groundwater moving along a lateral flowpath from a grass seed cropping system through an undisturbed mixed-species herbaceous riparian area. Soil A (30-45 cm) and C horizon (135-150 cm) NO3, dissolved oxygen, and nitrous oxide concentrations were significantly higher in the cropping system than the adjacent riparian area. Nitrate concentrations in both horizons of the riparian soil were consistently at or below 0.05 mg N L(-1) while cropping system concentrations ranged from 1 to 12 mg N L(-1). Chloride data suggested that NO3 dilution occurred from recharge by precipitation. However, a sharp decrease in NO3/Cl ratios as water moved into the riparian area indicated that additional dilution of NO3 concentrations was unlikely. Riparian area A horizon soil water had higher dissolved organic carbon than the cropping system and when the riparian soil became saturated, available electron acceptors (O2, NO3) were rapidly reduced. Dissolved inorganic carbon was significantly higher in the riparian area than the cropping system for both horizons indicating high biological activity. Carbon limitation in the cropping system may have led to microbial respiration using primarily O2 and to a lesser degree NO3. Within 6 m of the riparian/cropping system transition, NO3 was virtually undetectable. PMID- 17412900 TI - Soil microbial community response to hexavalent chromium in planted and unplanted soil. AB - Theories suggest that rapid microbial growth rates lead to quicker development of metal resistance. We tested these theories by adding hexavalent chromium [Cr(VI)] to soil, sowing Indian mustard (Brassica juncea), and comparing rhizosphere and bulk soil microbial community responses. Four weeks after the initial Cr(VI) application we measured Cr concentration, microbial biomass by fumigation extraction and soil extract ATP, tolerance to Cr and growth rates with tritiated thymidine incorporation, and performed community substrate use analysis with BIOLOG GN plates. Exchangeable Cr(VI) levels were very low, and therefore we assumed the Cr(VI) impact was transient. Microbial biomass was reduced by Cr(VI) addition. Microbial tolerance to Cr(VI) tended to be higher in the Cr-treated rhizosphere soil relative to the non-treated systems, while microorganisms in the Cr-treated bulk soil were less sensitive to Cr(VI) than microorganisms in the non treated bulk soil. Microbial diversity as measured by population evenness increased with Cr(VI) addition based on a Gini coefficient derived from BIOLOG substrate use patterns. Principal component analysis revealed separation between Cr(VI) treatments, and between rhizosphere and bulk soil treatments. We hypothesize that because of Cr(VI) addition there was indirect selection for fast growing organisms, alleviation of competition among microbial communities, and increase in Cr tolerance in the rhizosphere due to the faster turnover rates in that environment. PMID- 17412901 TI - Zinc accumulation in plant species indigenous to a Portuguese polluted site: relation with soil contamination. AB - The levels of zinc accumulated by roots, stems, and leaves of two plant species, Rubus ulmifolius and Phragmites australis, indigenous to the banks of a stream in a Portuguese contaminated site were investigated in field conditions. R. ulmifolius, a plant for which studies on phytoremediation potential are scarce, dominated on the right side of the stream, while P. australis proliferated on the other bank. Heterogeneous Zn concentrations were found along the banks of the stream. Zn accumulation in both species occurred mainly in the roots, with poor translocation to the aboveground sections. R. ulmifolius presented Zn levels in the roots ranging from 142 to 563 mg kg(-1), in the stems from 35 to 110 mg kg( 1), and in the leaves from 45 to 91 mg kg(-1), vs. average soil total Zn concentrations varying from 526 to 957 mg kg(-1). P. australis showed Zn concentrations in the roots from 39 to 130 mg kg(-1), in the stems from 31 to 63 mg kg(-1), and in the leaves from 37 to 83 mg kg(-1), for the lower average soil total Zn levels of 138 to 452 mg kg(-1) found on the banks where they proliferated. Positive correlations were found between the soil total, available and extractable Zn fractions, and metal accumulation in the roots and leaves of R. ulmifolius and in the roots and stems of P. australis. The use of R. ulmifolius and P. australis for phytoextraction purposes does not appear as an effective method of metal removing, but these native metal tolerant plant species may be used to reduce the effects of soil contamination, avoiding further Zn transfer to other environmental compartments. PMID- 17412902 TI - Assessment of contamination from arsenical pesticide use on orchards in the Great Valley region, Virginia and West Virginia, USA. AB - Lead arsenate pesticides were widely used in apple orchards from 1925 to 1955. Soils from historic orchards in four counties in Virginia and West Virginia contained elevated concentrations of As and Pb, consistent with an arsenical pesticide source. Arsenic concentrations in approximately 50% of the orchard site soils and approximately 1% of reference site soils exceed the USEPA Preliminary Remediation Goal (PRG) screening guideline of 22 mg kg(-1) for As in residential soil, defined on the basis of combined chronic exposure risk. Approximately 5% of orchard site soils exceed the USEPA PRG for Pb of 400 mg kg(-1) in residential soil; no reference site soils sampled exceed this value. A variety of statistical methods were used to characterize the occurrence, distribution, and dispersion of arsenical pesticide residues in soils, stream sediments, and ground waters relative to landscape features and likely background conditions. Concentrations of Zn, Pb, and Cu were most strongly associated with high developed land density and population density, whereas elevated concentrations of As were weakly correlated with high orchard density, consistent with a pesticide residue source. Arsenic concentrations in ground water wells in the region are generally <0.005 mg L(-1). There was no spatial association between As concentrations in ground water and proximity to orchards. Arsenic had limited mobility into ground water from surface soils contaminated with arsenical pesticide residues at concentrations typically found in orchards. PMID- 17412903 TI - Ground water stratification and delivery of nitrate to an incised stream under varying flow conditions. AB - Ground water processes affecting seasonal variations of surface water nitrate concentrations were investigated in an incised first-order stream in an agricultural watershed with a riparian forest in the coastal plain of Maryland. Aquifer characteristics including sediment stratigraphy, geochemistry, and hydraulic properties were examined in combination with chemical and isotopic analyses of ground water, macropore discharge, and stream water. The ground water flow system exhibits vertical stratification of hydraulic properties and redox conditions, with sub-horizontal boundaries that extend beneath the field and adjacent riparian forest. Below the minimum water table position, ground water age gradients indicate low recharge rates (2-5 cm yr(-1)) and long residence times (years to decades), whereas the transient ground water wedge between the maximum and minimum water table positions has a relatively short residence time (months to years), partly because of an upward increase in hydraulic conductivity. Oxygen reduction and denitrification in recharging ground waters are coupled with pyrite oxidation near the minimum water table elevation in a mottled weathering zone in Tertiary marine glauconitic sediments. The incised stream had high nitrate concentrations during high flow conditions when much of the ground water was transmitted rapidly across the riparian zone in a shallow oxic aquifer wedge with abundant outflow macropores, and low nitrate concentrations during low flow conditions when the oxic wedge was smaller and stream discharge was dominated by upwelling from the deeper denitrified parts of the aquifer. Results from this and similar studies illustrate the importance of near-stream geomorphology and subsurface geology as controls of riparian zone function and delivery of nitrate to streams in agricultural watersheds. PMID- 17412904 TI - Phosphorus transport through subsurface drainage and surface runoff from a flat watershed in east central Illinois, USA. AB - A long-term water quality monitoring program was established to evaluate the effects of agricultural management practices on water quality in the Little Vermilion River (LVR) watershed, IL. This watershed has intensive random and irregular subsurface drainage systems. The objective of this study was to assess the fate and transport of soluble phosphorus (soluble P) through subsurface drainage and surface runoff. Four sites (sites A, B, C, and E) that had subsurface and surface monitoring programs were selected for this study. Three of the four study sites had corn (Zea mays L.) and soybeans (Glycine max L.) planted in rotations and the other site had seed corn and soybeans. Subsurface drainage and surface runoff across all sites removed an average of 16.1 and 2.6% of rainfall, respectively. Annual flow-weighted soluble P concentrations fluctuated with the precipitation, while concentrations tended to increase with high precipitation coupled with high application rates. The long-term average flow weighted soluble P concentrations in subsurface flow were 102, 99, 194, and 86 microg L(-1) for sites A, B, C, and E, respectively. In contrast, the long-term average flow-weighted soluble P concentrations in surface runoff were 270, 253, 534, and 572 microg L(-1) for sites As, Bs, Cs, and Es, respectively. These values were substantially greater than the critical values that promote eutrophication. Statistical analysis indicated that the effects of crop, discharge, and the interactions between site and discharge and crop and discharge on soluble P concentrations in subsurface flow were significant (alpha = 0.05). Soluble P mass loads in surface runoff responded to discharge more consistently than in the subsurface flow. Subsurface flow had substantially greater annual average soluble P mass loads than surface runoff due to greater flow volume. PMID- 17412905 TI - Controls on catchment-scale patterns of phosphorus in soil, streambed sediment, and stream water. AB - Many models of phosphorus (P) transfer at the catchment scale rely on input from generic databases including, amongst others, soil and land use maps. Spatially detailed geochemical data sets have the potential to improve the accuracy of the input parameters of catchment-scale nutrient transfer models. Furthermore, they enable the assessment of the utility of available, generic spatial data sets for the modeling and prediction of soil nutrient status and nutrient transfer at the catchment scale. This study aims to quantify the unique and joint contribution of soil and sediment properties, land cover, and point-source emissions to the spatial variation of P concentrations in soil, streambed sediments, and stream water at the scale of a medium-sized catchment. Soil parent material and soil chemical properties were identified as major factors controlling the catchment scale spatial variation in soil total P and Olsen P concentrations. Soil type and land cover as derived from the generic spatial database explain 33.7% of the variation in soil total P concentrations and 17.4% of the variation in Olsen P concentrations. Streambed P concentrations are principally related to the major element concentrations in streambed sediment and P delivery from the hillslopes due to sediment erosion. During base flow conditions, the total phosphorus (<0.45 microm) concentrations in stream water are mainly controlled by the concentrations of P and the major elements in the streambed sediment. PMID- 17412906 TI - Occurrence and fate of the phytotoxin juglone in alley soils under black walnut trees. AB - Juglone (5-hydroxy-1,4-napthoquinone) is a chemical released by walnut trees, which can be toxic at various levels to several plant species. A balance among competing source and sink mechanisms and rates will ultimately determine whether juglone is capable of attaining sufficient levels to be allelopathic to intercrops in a walnut tree agroforestry system. In this study, juglone's release, accumulation, and decline in soil are explored using data from soil beneath a black walnut tree (Juglans nigra L) alley cropping system, greenhouse pot studies, and laboratory sorption/degradation studies. Juglone pore water concentrations estimated from extracts of surficial soil from beneath the alley cropping system exceeded the lowest solution culture toxicity levels reported for some plants of 10(-7) M, but did not exceed the inhibition threshold reported for typical intercrops such as maize and soybeans 10(-5) M. Further assessment of the likely persistence of juglone in soils indicated that juglone is both microbially and abiotically degraded, and that it will be particularly short-lived in soils supporting microbial activity. However, walnut seedlings planted in sand-filled pots clearly showed that juglone is released in measurable quantities to the soil's rhizosphere. Therefore, juglone accumulation in low fertility soils is plausible, and may still be worthy of consideration in management of alley agroforestry systems. PMID- 17412907 TI - Identifying fecal sources in a selected catchment reach using multiple source tracking tools. AB - Given known limitations of current microbial source-tracking (MST) tools, emphasis on small, simple study areas may enhance interpretations of fecal contamination sources in streams. In this study, three MST tools-Escherichia coli repetitive element polymerase chain reaction (rep-PCR), coliphage typing, and Bacteroidales 16S rDNA host-associated markers-were evaluated in a selected reach of Plum Creek in south-central Nebraska. Water-quality samples were collected from six sites. One reach was selected for MST evaluation based on observed patterns of E. coli contamination. Despite high E. coli concentrations, coliphages were detected only once among water samples, precluding their use as a MST tool in this setting. Rep-PCR classification of E. coli isolates from both water and sediment samples supported the hypothesis that cattle and wildlife were dominant sources of fecal contamination, with minor contributions by horses and humans. Conversely, neither ruminant nor human sources were detected by Bacteroidales markers in most water samples. In bed sediment, ruminant- and human associated Bacteroidales markers were detected throughout the interval from 0 to 0.3 m, with detections independent of E. coli concentrations in the sediment. Although results by E. coli-based and Bacteroidales-based MST methods led to similar interpretations, detection of Bacteroidales markers in sediment more commonly than in water indicates that different tools to track fecal contamination (in this case, tools based on Bacteroidales DNA and E. coli isolates) may have varying relevance to the more specific goal of tracking the sources of E. coli in watersheds. This is the first report of simultaneous, toolbox approach application of a library-based and marker-based MST analyses to flowing surface water. PMID- 17412908 TI - Phosphorus losses in simulated rainfall runoff from manured soils of Alberta. AB - Manure applied to agricultural land at rates that exceed annual crop nutrient requirements can be a source of phosphorus in runoff. Manure incorporation is often recommended to reduce phosphorus losses in runoff. A small plot rainfall simulation study was conducted at three sites in Alberta to evaluate the effects of manure rate and incorporation on phosphorus losses. Treatments consisted of three solid beef cattle manure application rates (50, 100, and 200 kg ha(-1) total phosphorus), an unmanured control, and two incorporation methods (nonincorporated and incorporated with one pass of a double disk). Simulated rain was applied to soils with freshly applied and residual (1 yr after application) manure at 70 mm h(-1) to produce 30 min of runoff. Soil test phosphorus (STP), total phosphorus (TP), and dissolved reactive phosphorus (DRP) concentrations in runoff increased with manure rate for fresh and residual manure. Initial abstraction and runoff volumes did not change with manure rate. Initial abstraction, runoff volumes, and phosphorus concentrations did not change with manure incorporation at Lacombe and Wilson, but initial abstraction volumes increased and runoff volumes and phosphorus concentrations decreased with incorporation of fresh manure at Beaverlodge. Phosphorus losses in runoff were directly related to phosphorus additions. Extraction coefficients (slopes of the regression lines) for the linear relationships between residual manure STP and phosphorus in runoff were 0.007 to 0.015 for runoff TP and 0.006 to 0.013 for runoff DRP. While incorporation of manure with a double disk had no significant effect on phosphorus losses in runoff from manure-amended soils 1 yr after application, incorporation of manure is still recommended to control nitrogen losses, improve crop nutrient uptake, and potentially reduce odor concerns. PMID- 17412909 TI - Flow-through rates and evaluation of solids separation of compost filter socks versus silt fence in sediment control applications. AB - Soil loss rates from construction sites can be 1000 times the average of natural soil erosion rates and 20 times that from agricultural lands. Silt fence (SF) is the current industry standard used to control sediment originating from construction activities. Silt fences are designed to act as miniature detention ponds. Research has indicated that SF sediment filtering efficiency is related to its ability to detain and pond water, not necessarily the filtration ability of the fabric. Design capacity and spacing is based on flow-through rate and design height. In addition, increased detention of runoff and pressure from ponding may increase the likelihood of overtopping or failure of SF in field application. Testing was conducted on compost silt socks (SS) and SF to determine sediment filtering efficiency, flow-through rate, ponding depth, overtopping point, design height, and design capacity. Results indicate flow-through rate changes with time, as does ponding depth, due to the accumulation of solids on/in the sediment filters. Changes in depth with time were a linear function of flow rate after 10 min of flow, up to the time the sediment filter is overtopped. Predicting the capacity of SF and SS to handle runoff without the filter being overtopped requires consideration of both runoff rate and length of runoff time. Data show SS half the heights of SF were less likely to overtop than SF when sediment-laden runoff water flow rates are less than 1.03 L(-1) s(-1) m(-1) (5 gpm/ft, gal per minute per lineal foot). Ponded depth behind a 61.0-cm (24 in) SF increased more rapidly than behind a 30.5-cm diam. (12 in) SS, and at the end of the thirty minutes, the depth behind the SF was 75% greater than that behind the SS. Removal of solids by the SF and the SS were not shown to be statistically different. Results were used to create a Microsoft Excel-based interactive design tool to assist engineers and erosion and sediment control planners on how to specify compost SS relative to SF in perimeter sediment control applications. PMID- 17412910 TI - Sorption and fractionation of dissolved organic matter and associated phosphorus in agricultural soil. AB - Mobility of dissolved organic matter (DOM) strongly affects the export of nitrogen (N) and phosphorus (P) from soils to surface waters. To study the sorption and mobility of dissolved organic C and P (DOC, DOP) in soil, the pH dependent sorption of DOM to samples from Ap, EB, and Bt horizons from a Danish agricultural Humic Hapludult was investigated and a kinetic model applicable in field-scale models tested. Sorption experiments of 1 to 72 h duration were conducted at two pH levels (pH 5.0 and 7.0) and six initial DOC concentrations (0 4.7 mmol L(-1)). Most sorption/desorption occurred during the first few hours. Dissolved organic carbon and DOP sorption decreased strongly with increased pH and desorption dominated at pH 7, especially for DOC. Due to fractionation during DOM sorption/desorption at DOC concentrations up to 2 mmol L(-1), the solution fraction of DOM was enriched in P indicating preferred leaching of DOP. The kinetics of sorption was expressed as a function of how far the solution DOC or DOP concentrations deviate from "equilibrium." The model was able to simulate the kinetics of DOC and DOP sorption/desorption at all concentrations investigated and at both pH levels making it useful for incorporation in field-scale models for quantifying DOC and DOP dynamics. PMID- 17412911 TI - Biosolids impact soil phosphorus accountability, fractionation, and potential environmental risk. AB - Biosolids land application rates are typically based on crop N requirements but can lead to soil P accumulation. The Littleton/Englewood, Colorado, wastewater treatment facility has supported biosolids beneficial-use on a dryland wheat fallow agroecosystem site since 1982, with observable soil P concentration increases as biyearly repeated biosolids applications increased from 0, 6.7, 13, 27, to 40 Mg ha(-1). The final study year was 2003, after which P accountability, fractionation, and potential environmental risk were assessed. Between 93 and 128% of biosolids-P added was accounted for when considering conventional tillage soil displacement, grain removal, and soil adsorption. The Fe-P fraction dominated all soil surface P fractions, likely due to an increase in amorphous Fe oxide because Fe2(SO4)3 was added at the wastewater treatment facility inflow for digester H2S reduction. The Ca-P phase dominated all soil subsurface P fractions due to calcareous soil conditions. A combination of conventional tillage, drought from 1999 to 2003, and repeated and increasing biosolids application rates may have forced soil surface microorganism dormancy, reduction, or mortality; thus, biomass P reduction was evident. Subsurface biomass P was greater than surface biomass, possibly due to protection against environmental and anthropogenic variables or to increased dissolved organic carbon inputs. Even given years of biosolids application, the soil surface had the ability to sorb additional P as determined by shaking the soil in an excessive P solution. Biosolids-application regulations based on the Colorado Phosphorus Index would not impede current site practices. Proper monitoring, management, and addition of other best management practices are needed for continued assurance that P movement off-site does not become a major issue. PMID- 17412912 TI - Influence of thymol and a urease inhibitor on coliform bacteria, odor, urea, and methane from a swine production manure pit. AB - Pathogens, ammonia, odor, and greenhouse gas emissions are serious environmental concerns associated with swine production. This study was conducted in two manure pits (33,000 L each) to determine the influence of 1.5 or 3.0 g thymol L(-1) and 80 mg L(-1) urease inhibitor amendments on urea accumulation, coliform bacteria, odor, and methane emission. Each experiment lasted 18 or 19 d, during which time 30 to 36 250-mL samples (six per day) were withdrawn from underneath each pit and analyzed for urea, thymol, volatile fatty acids, coliform bacteria, and Campylobacter. At the end of each experiment, six 50-g samples from each pit were placed in serum bottles, and gas volume and composition were determined periodically for 28 d. Compared with the control pit, volatile fatty acids production was reduced 64 and 100% for the thymol amendments of 1.5 and 3.0 g L( 1), respectively. Viable coliform cells were reduced 4.68 and 5.88 log10 colony forming units kg(-1) of slurry for the 1.5 and 3.0 g thymol L(-1), respectively, and Escherichia coli were reduced 4.67 and 5.01 log10 colony-forming units kg(-1) of slurry, respectively. Campylobacter was not detected in the pits treated with thymol, in contrast to 63% of the samples being positive for the untreated pit. Urea accumulated in the treated pits from Day 3 to 6. Total gas production from serum bottles was reduced 65 and 76% for thymol amendments of 1.5 and 3.0 g L( 1), respectively, and methane was reduced 78 and 93%, respectively. These results suggest that thymol markedly reduces pathogens, odor, and greenhouse gas emissions from a swine production facility. The urease inhibitor produced a temporary response in conserving urea. PMID- 17412913 TI - Hyperspectral reflectance response of freshwater macrophytes to salinity in a brackish subtropical marsh. AB - Coastal freshwater wetlands are threatened by increased salinity due to relative sea level rise and reduced freshwater inputs. Remote radiometric measurement of freshwater marsh canopies to detect small shifts in water column salinity would be useful for assessing salinity encroachment. We measured leaf hyperspectral (300-1100 nm) reflectance of freshwater macrophytes (cattail, Typha latifolia and sea oxeye, Borrichia frutescens) in a field study in a subtropical brackish (2.5 4.5 parts per thousand salinity, per thousand) marsh to determine salinity effects on visible and near-infrared spectral band reflectance and to identify reflectance indices sensitive to small (1 per thousand) changes in wetland salinity. For sea oxeye, floating-position water band index [fWBI = R(900)/minimum(R(930) - R(980)), where R(lambda) = reflectance at band lambda], normalized difference vegetation index [NDVI = (R(774) - R(681))/(R(774) + R(681))], and a proposed wetland salinity reflectance ratio (WSRR = R(990)/R(933)) were sensitive to salinity with R2 of 40, 35, and 65%, respectively (p < 0.01). For cattail, NDVI and photochemical reflectance index [PRI = (R(531) - R(570))/(R(570) + R(531))] were sensitive to salinity with R2 of 29 and 33%, respectively (p 17% total soluble protein) could be produced via transient expression: the protein assembled into higher-order structures visible by electron microscopy, and a concentrated extract was highly immunogenic in mice after subcutaneous injection and elicited high-titre neutralizing antibodies. Transgenic tobacco plants expressing a human codon-optimized gene linked to a chloroplast-targeting signal expressed L1 at levels up to 11% of the total soluble protein. These are the highest levels of HPV L1 expression reported for plants: these results, and the excellent immunogenicity of the product, significantly improve the prospects of making a conventional HPV vaccine by this means. PMID- 17412975 TI - Identification of HLA-DR1- and HLA-DR15-restricted human papillomavirus type 16 (HPV16) and HPV18 E6 epitopes recognized by CD4+ T cells from healthy young women. AB - Human papillomavirus (HPV) infection, particularly with types 16 and 18, is causally associated with the development of cervical cancer. Prophylactic vaccines against HPV have recently been licensed and have the primary aim of protecting children against future HPV infection and cervical cancer. However, these vaccines are unlikely to be effective in women with pre-existing HPV infection and disease. Previous studies have suggested that HPV16 E6-specific CD4+ T cells play a role in controlling viral infection; however, the epitopes recognized by such T-cells have not been defined. In this study, we analysed T cell responses against HPV16 and 18 in ten healthy young women in an age group (21-31) with a high prevalence of HPV infection and clearance. Five individuals made HPV E6 responses, from which five candidate T-cell epitopes (three HPV16 E6 and two HPV18 E6) were identified. More detailed characterization of epitopes from HPV16 E6(127-141) and HPV18 E6(43-57) revealed HLA-DRB1*01 and HLA-DRB1*15 restriction, respectively. Furthermore, generation of a T-cell line against HPV16 E6(127-141) demonstrated that this epitope could be presented after endogenous processing of soluble HPV16 E6 protein. Overall we demonstrate a powerful approach for defining novel CD4+ T-cell epitopes from two oncogenic HPV types. This approach could be applied to study populations in developing countries with a high incidence of cervical cancer. Such epitopes could provide a more precise way of investigating the role of natural and vaccine-induced T-cell responses against HPV in blood and at sites of disease. PMID- 17412976 TI - Characterization of two novel cutaneous human papillomaviruses, HPV93 and HPV96. AB - Two novel human papillomaviruses (HPVs), HPV93 and HPV96, with genomes of 7450 and 7438 bp, respectively, are described. The L1 open reading frame of HPV93 showed highest identity to HPV24 (79%) and that of HPV96 had highest identity to HPV92 (71%). Real-time PCR for HPV92, 93 and 96 on stripped biopsies from tumours and healthy skin from 269 immunocompetent patients found HPV DNA in 2.6% of tumours and in 0.4% of healthy skin samples. Double infections were observed in two tumours. HPV92 was detected in four, HPV93 in two and HPV96 in three tumours. The range of viral loads spanned from one copy per 45 cells to one copy per 10,000 cells. The E7 proteins of HPV92, 93 and 96 were found to bind the retinoblastoma protein (pRb). These results suggest a possible role for these HPV types in skin carcinogenesis that deserves further study. PMID- 17412977 TI - Complete genomic characterization of a murine papillomavirus isolated from papillomatous lesions of a European harvest mouse (Micromys minutus). AB - The papillomaviruses form a large group of species-specific pathogens that cause epithelial proliferations in a wide spectrum of animal hosts. Previous reports demonstrated a relatively high frequency of a variety of skin lesions in captive European harvest mice. The Micromys minutus papillomavirus (MmPV) was isolated from one of these lesions found on a captive European harvest mouse in a regional zoo in Chicago. In this study we present the entire genomic sequence of MmPV. The MmPV genome is organized into the seven classical papillomaviral open reading frames. Phylogenetic analysis places MmPV together with a papillomavirus (PV) isolated from a Syrian golden Hamster (HaOPV) in the genus Pipapillomavirus. The similar clustering pattern of the MmPV-HaOPV pair and their rodent hosts support the hypothesis of papillomaviral and host co-phylogenetic descent. The availability of the complete genomic sequence of a mouse PV should allow researchers to use MmPV as a model for PV carcinogenesis. PMID- 17412978 TI - Betapapillomaviruses frequently persist in the skin of healthy individuals. AB - Infections with human papillomaviruses (HPVs) belonging to the genus Betapapillomavirus have been linked to the development of non-melanoma skin cancer. Although persistence is expected, systematic investigation of this aspect of betapapillomavirus (beta-PV) infection has not been conducted. This study investigated the prevalence and persistence of 25 known beta-PV types in the skin of immunocompetent individuals. Over a 2 year period, eight consecutive plucked eyebrow hair samples taken from 23 healthy individuals were analysed for the presence of beta-PV DNA. Using a recently published general beta-PV PCR and genotyping method, 61% of the individuals were beta-PV DNA positive for one or more types at intake, whereas during follow-up this percentage rose to 96%. HPV23 was the most frequently detected beta-PV type. Type-specific beta-PV DNA was detected over 6 months or longer in 74% of the individuals. In 57% of the individuals, DNA from multiple beta-PV types was detected simultaneously for 6 months or longer. When the detection intervals of all beta-PV type-specific infections in the study population were considered, a substantial proportion, 48%, lasted at least half a year. The consistent beta-PV patterns found over time in most individuals strongly suggested that beta-PV DNA detection in plucked eyebrow hairs reveals true beta-PV infection. If the minimum interval of detection was set at 6 months, persistent beta-PV infections were found in the majority of the study population (74%). PMID- 17412979 TI - Identification and functional analysis of the origins of DNA replication in the Cydia pomonella granulovirus genome. AB - The entire genome of Cydia pomonella granulovirus (CpGV) was systematically screened for origins of DNA replication, using an infection-dependent DNA replication assay in the granulovirus-permissive Cydia pomonella cell line, Cp14R. All seven cosmids in an overlapping library that covered the CpGV genome were found to replicate in the assay. A genomic library of 32 overlapping plasmids was subsequently screened. Plasmids that replicated were in turn subcloned into 1-2 kbp overlapping fragments. Eleven subclones replicated, each containing at least one of the 13 single-copy 74-76 bp imperfect palindromes, previously identified in the CpGV genome as possible origins of replication. Genome fragments of 156 bp, each containing one of the 13 palindromes, were cloned to verify replication and provided confirmation that these 13 palindromes are the only origins of replication in the genome. A real-time PCR method was developed for the quantification of DNA replication, which eliminated the need for Southern blotting and hybridization. A set of deletion clones allowed further quantitative characterization of one of the palindromes. The previously proposed non-homologous region origin of replication did not replicate in the assay. PMID- 17412980 TI - Complete genomes for hepatitis C virus subtypes 6f, 6i, 6j and 6m: viral genetic diversity among Thai blood donors and infected spouses. AB - In this study, the first complete genome sequences for hepatitis C virus (HCV) subtypes 6f, 6i, 6j and 6m, obtained from infected blood donors in Chiang Mai, Thailand, are reported. Pairwise genome-wide nucleotide similarities between some of these isolates were higher than the 75-80% value used previously to define different HCV subtypes. To investigate further, the entire genomes of four prototype isolates, Th602 (6i), Th553 (6j), B4/92 (6m) and D86/93 (6n), were sequenced. Pairwise comparison of these sequences gave a similar range of nucleotide similarities, thereby providing new information for HCV subtype classification. In order to study the hypothesis of interspousal HCV transmission, four additional complete HCV genome sequences were obtained from two infected Thai blood donors and their spouses, C-0044 and C-0046 (6f), and C 0192 and C-0185 (6m). Pairwise comparison of the sequences revealed that C-0044 and C-0046 share a nucleotide similarity of 98.1%, whilst C-0185 and C-0192 have a similarity of 97.8%. Several other studies of partial HCV sequences of different genomic regions from HCV-infected couples have shown nucleotide similarities ranging from 96.3 to 100%. The similarities of the complete genome sequences from the two couples in the current study are consistent with HCV transmission between spouses. PMID- 17412981 TI - Complete genomes of hepatitis C virus (HCV) subtypes 6c, 6l, 6o, 6p and 6q: completion of a full panel of genomes for HCV genotype 6. AB - Five hepatitis C virus (HCV) complete genome sequences (Th846, 537796, QC227, QC216 and QC99) from a blood donor in Thailand and three Asian immigrants and one Caucasian in North America were determined. Phylogenetically, they represent the first complete genomes for subtypes 6c, 6l, 6o, 6p and 6q, respectively. Similarity analysis showed no evidence of inter- or intrasubtype recombination. Further analysis in conjunction with partial sequences from the Los Alamos HCV database led to the identification of other closely related isolates from south eastern Asia or immigrants from that region. However, Th846 did not cluster with any reference sequence and is the sole isolate of subtype 6c reported so far. This study completes the full genome sequencing of all 17 assigned HCV genotype 6 subtypes (6a-6q). The utility of this panel of complete sequences for accurate detection and classification of infection, and for estimating the origin of this genotype of HCV, is discussed. PMID- 17412982 TI - Genetic diversity in hepatitis C virus in Egypt and possible association with hepatocellular carcinoma. AB - Egypt has one of the world's highest prevalences of hepatitis C virus (HCV) infection, with a majority of genotype 4 infections. To explore the genetic diversity of HCV in Egypt, sera from 131 Egyptians [56 from community studies, 37 chronic hepatitis patients, 28 hepatocellular carcinoma (HCC) patients and 10 patients with non-Hodgkin's lymphoma] were genotyped by restriction fragment length polymorphism and phylogenetic analyses of sequences from the mid-core and non-structural 5B regions. The different genotyping methods showed good agreement. The majority of the viruses (83 of 131; 63%) were of subtype 4a, but five other subtypes within genotype 4 were also observed, as well as three genotype 1b, five genotype 1g and one genotype 3a samples. Interestingly, subtype 4o, which was easily identifiable in all three genomic regions, showed an association with HCC (P=0.017), which merits further investigation. PMID- 17412983 TI - Egg whites from eggs of chickens infected experimentally with avian hepatitis E virus contain infectious virus, but evidence of complete vertical transmission is lacking. AB - Avian hepatitis E virus (HEV) is genetically and antigenically related to human HEV. Vertical transmission of HEV has been reported in humans, but not in other animals. In this study, we showed that avian HEV could be detected in chicken egg white samples. Subsequently, avian HEV in egg white was found to be infectious, as evidenced by the appearance of viraemia, faecal virus shedding and seroconversion in chickens inoculated with avian HEV-positive egg white, but not in chickens inoculated with HEV-negative egg white. To further assess the possibility of vertical transmission of avian HEV, batches of embryonated eggs from infected hens were hatched, and hatched chicks were monitored for evidence of avian HEV infection. However, no virus was detected in samples collected from the hatched chicks throughout this study, suggesting that avian HEV could not complete the vertical transmission cycle. The possible implications of our findings are also discussed. PMID- 17412984 TI - Analysis of the complete genomic sequence of an apparently avirulent strain of avian hepatitis E virus (avian HEV) identified major genetic differences compared with the prototype pathogenic strain of avian HEV. AB - Avian hepatitis E virus (HEV) was identified from chickens with hepatitis splenomegaly syndrome. In this study, the complete genomic sequence of an apparently avirulent strain of avian HEV was determined to be 6649 nt in length, excluding the poly(A) tail, which is 5 nt shorter than the prototype avian HEV. Sequence analyses revealed that the ORF1 has 89.6% nucleotide sequence identity, with numerous non-silent mutations and deletions, compared to the prototype strain. The ORF2 capsid gene showed 90.7% sequence identity with six non-silent mutations, and ORF3 had four non-silent mutations with 97% sequence identity. Overall, the apparently avirulent strain shares only 90.1% nucleotide sequence identity with the prototype strain. The identification of significant non-silent mutations in the capsid gene and other regions suggests that these mutations may play a role in HEV attenuation. This is the first report of the full-length sequence of an apparently avirulent strain of HEV. PMID- 17412985 TI - Genetic determinants of Sindbis virus strain TR339 affecting midgut infection in the mosquito Aedes aegypti. AB - Mosquito midgut epithelial cells (MEC) play a major role in determining whether an arbovirus can successfully infect and be transmitted by mosquitoes. The Sindbis virus (SINV) strain TR339 efficiently infects Aedes aegypti MEC but the SINV strain TE/5'2J poorly infects MEC. SINV determinants for MEC infection have been localized to the E2 glycoprotein. The E2 amino acid sequences of TR339 and TE/5'2J differ at two sites, E2-55 and E2-70. We have altered the TE/5'2J virus genome by site-directed mutagenesis to contain two TR339 residues, E2-55 H-->Q (histidine to glutamine) and E2-70 K-->E (lysine to glutamic acid). We have characterized the growth patterns of derived viruses in cell culture and determined the midgut infection rate (MIR) in A. aegypti mosquitoes. Our results clearly show that the E2-55 H-->Q and the E2-70 K-->E mutations in the TE/5'2J virus increase MIR both independently and in combination. TE/5'2J virus containing both TR339 E2 residues had MIRs similar to the parental TR339 virus. In addition, SINV propagated in a mammalian cell line had a significantly lower A. aegypti midgut 50 % infectious dose than virus propagated in a mosquito cell line. PMID- 17412986 TI - Further characterization of a paramyxovirus transcription initiation signal: search for required nucleotides upstream and importance of the N phase context. AB - Paramyxovirus genomes contain a linear array of five to ten genes sequentially transcribed by the viral RNA polymerase. mRNA synthesis initiates at a nucleotide signal (gs1) within the genomic promoter located at the genome 3' end. To gain information about the mechanism involved in transcription initiation, a search was carried out for upstream nucleotides required for gs1 and the effects of the gs1 nucleocapsid protein (N) phase context on transcription regulation were determined. For both purposes, tandem promoter mini-genomes carrying a transcription signal ectopically positioned downstream of a replication-only signal were used. The requirement for hygromycin resistance gene expression was used in an attempt to select essential nucleotides within randomized stretches of nucleotides. Nucleotide insertions or deletions were also made on either side of the transcription signal to change its original N phase context in the five remaining possibilities and GFP expression from these modified signals was assessed. Cell cultures resistant to hygromycin treatment were readily obtained following amplification of mini-genomes harbouring randomized sequences. However, selected nucleotides upstream of gs1 could not be identified under conditions where nucleotides within gs1 were selected. In contrast, it was observed that changing the gs1 N phase context progressively decreased transcription by five- to tenfold. These results are discussed in relation to two different mechanisms of transcription initiation. PMID- 17412987 TI - A novel monolayer cell line derived from human umbilical cord blood cells shows high sensitivity to measles virus. AB - Measles virus (MeV) research is largely dependent on the B95a cell line, which is derived from marmoset B lymphocytes. As this cell line is persistently infected with Epstein-Barr virus (EBV), a novel cell line, COBL-a, was established from human umbilical cord blood. COBL-a cells have a significant advantage over B95a cells because they are of human origin, are free from EBV and have higher sensitivity to wild-type MeV. Thus, COBL-a cells should prove very valuable for both epidemiological and basic studies of MeV. PMID- 17412988 TI - Analysis of gene expression in Lassa virus-infected HuH-7 cells. AB - The pathogenesis of Lassa fever is poorly understood. As the liver is a major target organ of Lassa virus, gene expression in Lassa virus-infected HuH-7 cells, a differentiated human hepatoma cell line, was studied. Cellular mRNA levels were measured at the late phase of acute infection, when virtually all cells expressed large amounts of nucleoprotein, and virus RNA concentration had reached>10(8) copies (ml supernatant)-1. Two types of transcription array were used: cDNA-based macroarrays with a set of 3500 genes (Atlas Human 1.2 arrays; Clontech) and oligonucleotide-based microarrays covering 18,400 transcripts (Human Genome U133A array; Affymetrix). Data analysis was based on statistical frameworks controlling the false-discovery rate. Atlas array data were considered relevant if they could be verified by U133A array or real-time RT-PCR. According to these criteria, there was no evidence for true changes in gene expression. Considering the precision of the U133A array and the number of replicates tested, potential expression changes due to Lassa virus infection are probably smaller than twofold. To substantiate the array data, beta interferon (IFN-beta) gene expression was studied longitudinally in Lassa virus-infected HuH-7 and FRhK-4 cells by using real-time RT-PCR. IFN-beta mRNA levels increased only twofold upon Lassa virus infection, although there was no evidence that the virus inhibited poly(I:C)-induced IFN-beta gene expression. In conclusion, Lassa virus interferes only minimally with gene expression in HuH-7 cells and poorly induces IFN-beta gene transcription. PMID- 17412989 TI - Induction of apoptosis in Saccharomyces cerevisiae results in the spontaneous maturation of tetravirus procapsids in vivo. AB - The Tetraviridae are a family of small, non-enveloped, insect RNA viruses consisting of one or two single-stranded, positive-sense genomic RNAs encapsidated in an icosahedral capsid with T=4 symmetry. Tetravirus procapsids undergo maturation when exposed to a low pH environment in vitro. While the structural biology of the conformational changes that mediate acid-dependent maturation is well understood, little is known about the significance of acid dependent maturation in vivo. To address this question, the capsid-coding sequence of the tetravirus Helicoverpa armigera stunt virus was expressed in Saccharomyces cerevisiae cells. Virus-like particles were shown to assemble as procapsids that matured spontaneously in vivo as the cells began to age. Growth in the presence of hydrogen peroxide or acetic acid, which induced apoptosis or programmed cell death in the yeast cells, resulted in virus-like particle maturation. The results demonstrate that assembly-dependent maturation of tetravirus procapsids in vivo is linked to the onset of apoptosis in yeast cells. We propose that the reduction in pH required for tetraviral maturation may be the result of cytosolic acidification, which is associated with the early onset of programmed cell death in infected cells. PMID- 17412990 TI - Identification of minimal sequences of the Rhopalosiphum padi virus 5' untranslated region required for internal initiation of protein synthesis in mammalian, plant and insect translation systems. AB - Rhopalosiphum padi virus (RhPV) is a member of the family Dicistroviridae. The genomes of viruses in this family contain two open reading frames, each preceded by distinct internal ribosome entry site (IRES) elements. The RhPV 5' IRES is functional in mammalian, insect and plant translation systems and can form 48S initiation complexes in vitro with just the mammalian initiation factors eIF2, eIF3 and eIF1. Large regions of the 5' untranslated region (UTR) can be deleted without affecting initiation-complex formation. The minimal sequences required for directing internal initiation in mammalian (rabbit reticulocyte lysate), plant (wheatgerm extract) and insect (Sf21 cells) translation systems have now been defined. A fragment (nt 426-579) from the 3' portion of the 5' UTR can direct translation in each of these translation systems. In addition, a distinct region (nt 300-429) is also active. Thus, unstructured regions within the 5' UTR seem to be critical for IRES function. PMID- 17412991 TI - Vaccination with a T-cell-priming Gag peptide of caprine arthritis encephalitis virus enhances virus replication transiently in vivo. AB - CD4+ T cells are involved in several immune response pathways used to control viral infections. In this study, a group of genetically defined goats was immunized with a synthetic peptide known to encompass an immunodominant helper T cell epitope of caprine arthritis encephalitis virus (CAEV). Fifty-five days after challenge with the molecularly cloned CAEV strain CO, the vaccinated animals had a higher proviral load than the controls. The measurement of gamma interferon and interleukin-4 gene expression showed that these cytokines were reliable markers of an ongoing immune response but their balance did not account for more or less efficient control of CAEV replication. In contrast, granulocyte macrophage colony-stimulating factor appeared to be a key cytokine that might support virus replication in the early phase of infection. The observation of a potential T-cell-mediated enhancement of virus replication supports other recent findings showing that lentivirus-specific T cells can be detrimental to the host, suggesting caution in designing vaccine candidates. PMID- 17412992 TI - Molecular dissection of the potato virus Y VPg virulence factor reveals complex adaptations to the pvr2 resistance allelic series in pepper. AB - The virulence properties of potato virus Y (PVY) towards an allelic series at the pvr2 locus in pepper genotypes are related to variations in the genome-linked viral protein (VPg). Eleven amino acid substitutions in the central part of the VPg were identified in strains differing by their virulence properties and were introduced, either singly or in combination, in an infectious PVY clone to get an in-depth genetic analysis of the virulence determinant. The virulence spectrum of these mutants was evaluated by inoculation of four pepper genotypes carrying different alleles at the pvr2 locus. The mutations introduced had complex effects on virulence, including antagonistic epistasis and trade-offs for virulence towards different pvr2 alleles. In addition, several mutants showed new virulence properties that were unknown in the natural environment. Such complex effects of mutations on plant virus virulence are unprecedented. They provide a better understanding of the variable levels of durability of the resistance conferred by the different pvr2 alleles, and have important consequences for a durable management of the resistances. PMID- 17412993 TI - In vitro association between the helper component-proteinase of zucchini yellow mosaic virus and cuticle proteins of Myzus persicae. AB - Potyviruses, as typical non-persistently transmitted viruses, are carried within the stylets of aphids. Cuticle proteins (CuPs), which are a major component of the insect cuticle, were examined for in vitro binding to the potyviral helper component-proteinase (HC-Pro). Proteins in 8 M urea extracts from Myzus persicae were separated by SDS-PAGE, electroblotted onto membranes and identified as CuPs by using specific antibodies to M. persicae CuP. Blotted M. persicae protein extracts were overlaid with two HC-Pros, differing by the presence of K or E in the KLSC domain. The HC-Pro with KLSC, known to assist transmission, was found to bind M. persicae proteins, whereas the HC-Pro with ELSC, being deficient in assisting transmission, did not. To identify CuPs that react with HC-Pro, protein extracts were separated by two-dimensional gel electrophoresis. Nine proteins reacting with HC-Pro were sequenced by mass spectrometry. Sequences of peptides in four proteins, of molecular masses between 22 and 31 kDa, were identified as CuPs according to comparison with sequences in GenBank. The putative CuPs from M. persicae that bind HC-Pro are potentially of interest in locating receptors for virions bound to HC-Pro in aphids' stylets. PMID- 17412994 TI - RNA4-encoded p31 of beet necrotic yellow vein virus is involved in efficient vector transmission, symptom severity and silencing suppression in roots. AB - RNA3 and RNA4 of beet necrotic yellow vein virus (BNYVV) are not essential for virus multiplication, but are associated with vector-mediated infection and disease development in sugar beet roots. Here, a unique role for RNA4 in virus transmission, virulence and RNA silencing suppression was demonstrated. Mutagenic analysis revealed that the RNA4-encoded p31 open reading frame (ORF) was involved in efficient vector transmission and slight enhancement of symptom expression in some Beta species. No effects of RNA4 on virus accumulation in infected tissue were observed. Furthermore, the p31 ORF was involved in the induction of severe symptoms by BNYVV in Nicotiana benthamiana plants without affecting viral RNA accumulation. In contrast, RNA3-encoded p25, previously identified as a major contributor to symptom induction in sugar beet, had no such effect on N. benthamiana. In two different silencing suppression assays, neither p31 nor p25 was able to suppress RNA silencing in leaves, but the presence of p31 enhanced a silencing suppressor activity in roots without alteration in viral RNA accumulation. Thus, BNYVV p31 plays a multifunctional role in efficient vector transmission, enhanced symptom expression and root-specific silencing suppression. PMID- 17412995 TI - A novel cleavage site within the potato leafroll virus P1 polyprotein. AB - To study the proteolytic processing of the potato leafroll virus replicase proteins, the multidomain P1 protein with a c-myc epitope tag attached at the N terminus was expressed in insect cells by using the baculovirus system. Western blotting showed that P1 was cleaved at a site upstream of the serine protease domain, in addition to the cleavage site downstream of the protease domain. Mutational analysis showed that the serine protease domain within P1 was responsible for this cleavage. To characterize this novel cleavage site further, a portion of the P1 protein comprising the protease domain and the two cleavage sites was expressed in Escherichia coli. A similar cleavage event was observed in bacteria and was abolished when the P1 protease was inactivated by mutation. Peptide-sequencing studies indicated that this cleavage occurred at a Glu/Arg junction, separating the N-terminal 204 residues from the serine protease domain of P1. PMID- 17412996 TI - Infectivity, pseudorecombination and mutagenesis of Kenyan cassava mosaic begomoviruses. AB - Cloned DNA-A and DNA-B components of Kenyan isolates of East African cassava mosaic virus (EACMV, EACMV-UG and EACMV-KE2), East African cassava mosaic Kenya virus (EACMKV) and East African cassava mosaic Zanzibar virus (EACMZV) are shown to be infectious in cassava. EACMV and EACMKV genomic components have the same iteron sequence (GGGGG) and can form viable pseudorecombinants, while EACMZV components have a different sequence (GGAGA) and are incompatible with EACMV and EACMKV. Mutagenesis of EACMZV has demonstrated that open reading frames (ORFs) AV1 (encoding the coat protein), AV2 and AC4 are not essential for a symptomatic infection of cassava, although mutants of both ORF AV1 and AV2 produce attenuated symptoms in this host. Furthermore, ORF AV1 and AV2 mutants were compromised for coat protein production, suggesting a close structural and/or functional relationship between these coding regions or their protein products. PMID- 17412997 TI - Cymbidium ringspot virus defective interfering RNA replication in yeast cells occurs on endoplasmic reticulum-derived membranes in the absence of peroxisomes. AB - The replication of Cymbidium ringspot virus (CymRSV) defective interfering (DI) RNA in cells of the yeast Saccharomyces cerevisiae normally takes place in association with the peroxisomal membrane, thus paralleling the replication events in infected plant cells. However, previous results with a peroxisome deficient mutant strain of yeast had suggested that the presence of peroxisomes is not a strict requirement for CymRSV DI RNA replication. Thus, a novel approach was used to study the putative alternative sites of replication by using S. cerevisiae strain YPH499 which does not contain normal peroxisomes. In this strain, CymRSV p33 and p92 accumulated over portions of the nuclear membrane and on membranous overgrowths which were identified as endoplasmic reticulum (ER) strands, following immunofluorescence and immunoelectron microscope observations. The proteins were not released by high-pH treatment, but were susceptible to proteolytic digestion, thus indicating peripheral and not integrated association. ER-associated p33 and p92 proteins supported in trans the replication of DI RNA. The capacity of plus-strand RNA viruses to replicate in association with different types of cell membranes was thus confirmed. PMID- 17412998 TI - Identification of vasodilator-stimulated phosphoprotein (VASP) as an HIF regulated tissue permeability factor during hypoxia. AB - Increased tissue permeability is commonly associated with hypoxia of many origins. Since hypoxia-inducible factor (HIF) represents a predominant hypoxia signaling mechanism, we compared hypoxia-elicited changes in tissue barrier function in mice conditionally lacking intestinal epithelial hypoxia-inducible factor-1alpha (hif1a). Somewhat surprisingly, these studies revealed that mutant hif1a mice were protected from hypoxia-induced increases in intestinal permeability in vivo. Guided by microarray analysis of tissues derived from these mutant hif1a mice, we identified HIF-1-dependent repression of vasodilator stimulated phosphoprotein (VASP), a molecule known to be important in the control of cytoskeletal dynamics, including barrier function. Studies at the mRNA and protein level confirmed hypoxia-elicited repression of VASP in murine tissue, cultured epithelia and endothelia, as well as human saphenous vein ex vivo. Targeted repression of VASP by siRNA recapitulated our findings with hypoxia and directed overexpression of VASP abolished hypoxia-induced barrier dysfunction. Studies in the cloned human VASP promoter revealed hypoxia-dependent transcriptional repression, and functional studies by chromatin immunoprecipitation (ChIP) and site-directed mutagenesis revealed hypoxia dependent binding of HIF-1alpha to the human VASP promoter. These studies identify HIF-1-dependent repression of VASP as a control point for hypoxia regulated barrier dysfunction. PMID- 17412999 TI - The extracellular chaperone clusterin influences amyloid formation and toxicity by interacting with prefibrillar structures. AB - Clusterin is an extracellular chaperone present in all disease-associated extracellular amyloid deposits, but its roles in amyloid formation and protein deposition in vivo are poorly understood. The current study initially aimed to characterize the effects of clusterin on amyloid formation in vitro by a panel of eight protein substrates. Two of the substrates (Alzheimer's beta peptide and a PI3-SH3 domain) were then used in further experiments to examine the effects of clusterin on amyloid cytotoxicity and to probe the mechanism of clusterin action. We show that clusterin exerts potent effects on amyloid formation, the nature and extent of which vary greatly with the clusterin:substrate ratio, and provide evidence that these effects are exerted via interactions with prefibrillar species that share common structural features. Proamyloidogenic effects of clusterin appear to be restricted to conditions in which the substrate protein is present at a very large molar excess; under these same conditions, clusterin coincorporates with substrate protein into insoluble aggregates. However, when clusterin is present at much higher but still substoichiometric levels (e.g., a molar ratio of clusterin:substrate=1:10), it potently inhibits amyloid formation and provides substantial cytoprotection. These findings suggest that clusterin is an important element in the control of extracellular protein misfolding. PMID- 17413000 TI - Disruption of androgen receptor signaling by synthetic progestins may increase risk of developing breast cancer. AB - There is now considerable evidence that using a combination of synthetic progestins and estrogens in hormone replacement therapy (HRT) increases the risk of breast cancer compared with estrogen alone. Furthermore, the World Health Organization has recently cited combination contraceptives, which contain synthetic progestins, as potentially carcinogenic to humans, particularly for increased breast cancer risk. Given the above observations and the current trend toward progestin-only contraception, it is important that we have a comprehensive understanding of how progestins act in the millions of women worldwide who regularly take these medications. While synthetic progestins, such as medroxyprogesterone acetate (MPA), which are currently used in both HRT and oral contraceptives were designed to act exclusively through the progesterone receptor, it is clear from both clinical and experimental settings that their effects may be mediated, in part, by binding to the androgen receptor (AR). Disruption of androgen action by synthetic progestins may have serious deleterious side effects in the breast, where the balance between estrogen signaling and androgen signaling plays a critical role in breast homeostasis. Here, we review the role of androgen signaling in the normal breast and in breast cancer and present new data demonstrating that androgen receptor function can be perturbed by low doses of MPA, similar to doses achieved in serum of women taking HRT. We propose that the observed excess of breast malignancies associated with combined HRT may be explained, in part, by synthetic progestins such as MPA acting as endocrine disruptors to negate the protective effects of androgen signaling in the breast. Understanding the role of androgen signaling in the breast and how this is modulated by synthetic progestins is necessary to determine how combined HRT alters breast cancer risk, and to inform the development of optimal preventive and treatment strategies for this disease. PMID- 17413001 TI - Aging perturbs 26S proteasome assembly in Drosophila melanogaster. AB - Aging is associated with loss of quality control in protein turnover. The ubiquitin-proteasome pathway is critical to this quality control process as it degrades mutated and damaged proteins. We identified a unique aging-dependent mechanism that contributes to proteasome dysfunction in Drosophila melanogaster. Our studies are the first to show that the major proteasome form in old (43-47 days old) female and male flies is the weakly active 20S core particle, while in younger (1-32 days old) flies highly active 26S proteasomes are preponderant. Old (43-47 days) flies of both genders also exhibit a decline (approximately 50%) in ATP levels, which is relevant to 26S proteasomes, as their assembly is ATP dependent. The steep declines in 26S proteasome and ATP levels were observed at an age (43-47 days) when the flies exhibited a marked drop in locomotor performance, attesting that these are "old age" events. Remarkably, treatment with a proteasome inhibitor increases ubiquitinated protein levels and shortens the life span of old but not young flies. In conclusion, our data reveal a previously unknown mechanism that perturbs proteasome activity in "old-age" female and male Drosophila most likely depriving them of the ability to effectively cope with proteotoxic damages caused by environmental and/or genetic factors. PMID- 17413002 TI - Medulloblastomas derived from Cxcr6 mutant mice respond to treatment with a smoothened inhibitor. AB - The sonic hedgehog (Shh) pathway is activated in approximately 30% of human medulloblastoma resulting in increased expression of downstream target genes. In about half of these cases, this has been shown to be a consequence of mutations in regulatory genes within the pathway, including Ptc1, Smo, and Sufu. However, for some tumors, no mutations have been detected in known pathway genes. This suggests that either mutations in other genes promote tumorigenesis or that epigenetic alterations increase pathway activity in these tumors. Here, we report that 3% to 4% of mice lacking either one or both functional copies of Cxcr6 develop medulloblastoma. Although CXCR6 is not known to be involved in Shh signaling, tumors derived from Cxcr6 mutant mice expressed Shh pathway target genes including Gli1, Gli2, Ptc2, and Sfrp1, indicating elevated pathway activity. Interestingly, the level of Ptc1 expression was decreased in tumor cells although two normal copies of Ptc1 were retained. This implies that reduced CXCR6 function leads to suppression of Ptc1 thereby increasing Smoothened function and promoting tumorigenesis. We used a direct transplant model to test the sensitivity of medulloblastoma arising in Cxcr6 mutant mice to a small molecule inhibitor of Smoothened (HhAntag). We found that transplanted tumors were dramatically inhibited in mice treated for only 4 days with HhAntag. These findings suggest that HhAntag may be effective against tumors lacking mutations in known Shh pathway genes. PMID- 17413003 TI - Cancer immunologists and cancer biologists: why we didn't talk then but need to now. AB - What is cancer? Cancer is a disease initiated by a series of cumulative genetic and epigenetic changes that occur in a normal cell. However, in addition to the malignant cell itself, cancer is a disease of microenvironment and immunity. Although genetic and epigenetic alterations drive cellular transformation, genomic plasticity, and evolution, it has become increasingly apparent that multiple signals delivered within the tumor microenvironment by modifier genes, stromal and endothelial cells, and immune cells are critical factors in determining the progression versus dormancy or destruction of an initiated lesion and also whether metastasis may occur. With regard to the important roles of immune cells in cancer, a chasm exists between immunologists and biologists: although sharing a common disease interest, there is little history for workers to draw on based on shared perspectives or understanding. How did this disconnect arise? Here, we look at how these workers became separated in the past and address why it has now become critical to spur greater cross-fertilization. In particular, we highlight three ideas that we believe are important for discussion and debate. The first idea is that therapeutic strategies that fail to harness the immune system will always be defeated by tumor resistance, due to the large "genomic space" that genetically plastic tumor cells can readily access to evolve resistance mechanisms. Because all therapies drive tumor progression by imposing a selection for resistant cells, harnessing the adaptivity of the immune system will be indispensable to ultimately stanching the deadly adaptability of the tumor cell. The second idea is that using molecular targeted agents to reverse tumoral immune suppression may offer a powerful method to leverage the efficacy of most if not all therapeutic agents. We suggest that the mechanisms that support evolution of a "smoldering" inflammatory environment in cancer overlap with those that support evolution of tumoral immune escape. If true, relieving immune suppression will switch the inflammatory state from supportive to destructive for the tumor. The third idea is that by ablating immunosuppression mechanisms, cytotoxic chemotherapy might synergize with, rather than antagonize, active immunotherapy. Provocative preclinical studies in this area prompt clinical attention. We believe that increased efforts to intermingle the perspectives and work of cancer immunologists with cancer biologists and pharmacologists will be needed to realize the National Cancer Institute's goal of managing cancer in the clinic by 2015. PMID- 17413005 TI - Making new memories: the role of the hippocampus in new associative learning. AB - Both aging and Alzheimer's disease target the hippocampal formation and can result in mild to devastating memory impairment depending on the severity of the condition. Understanding the normal mnemonic functions of the hippocampus and related structures of the medial temporal lobe is the first step toward the development of diagnostics and treatments designed to ameliorate these potentially devastating age-related memory deficits. Here I describe findings from behavioral neurophysiological studies in which we have investigated the patterns of dynamic neural activity seen in the macaque monkey hippocampus during the acquisition of new associative memories. We report that hippocampal neurons signal the formation of new associations with dramatic changes in their firing rate. Because these learning-related signals can occur just before behavioral learning is expressed, this suggests that these signals play a role in driving the learning process. Implications of these findings for understanding the memory deficits associated with aging and Alzheimer's disease are discussed. PMID- 17413006 TI - Anatomical and functional phenotyping of mice models of Alzheimer's disease by MR microscopy. AB - The wide variety of transgenic mouse models of Alzheimer's disease (AD) reflects the search for specific genes that influence AD pathology and the drive to create a clinically relevant animal model. An ideal AD mouse model must display hallmark AD pathology such as amyloid plaques, neurofibrillary tangles, reactive gliosis, dystrophic neurites, neuron and synapse loss, and brain atrophy and in parallel behaviorally mimic the cognitive decline observed in humans. Magnetic resonance (MR) microscopy (MRM) can detect amyloid plaque load, development of brain atrophy, and acute neurodegeneration. MRM examples of AD pathology will be presented and discussed. What has lagged behind in preclinical research using transgenic AD mouse models is functional phenotyping of the brain; in other words, the ability to correlate a specific genotype with potential aberrant brain activation patterns. This lack of information is caused by the technical challenges involved in performing functional MRI (fMRI) in mice including the effects of anesthetic agents and the lack of relevant "cognitive" paradigms. An alternative approach to classical fMRI using external stimuli as triggers of brain activation in rodents is to electrically or pharmacologically stimulate regions directly while simultaneously locally tracking the activated interconnected regions of rodents using, for example, the manganese-enhanced MRI (MEMRI) technique. Finally, transgenic mouse models, MRM, and future AD research would be strengthened by the ability to screen for AD-like pathology in other non AD transgenic mouse models. For example, molecular biologists may focus on cardiac or pulmonary pathologies in transgenic mice models and as an incidental finding discover behavioral AD phenotypes. We will present MRM data of brain and cardiac phenotyping in transgenic mouse models with behavioral deficits. PMID- 17413007 TI - Various dendritic abnormalities are associated with fibrillar amyloid deposits in Alzheimer's disease. AB - Dystrophic neurites are associated with fibrillar amyloid deposition in Alzheimer's disease (AD), but the frequency and types of changes in synaptic structures near amyloid deposits have not been well characterized. Using high resolution confocal microscopy to image lipophilic dye-labeled dendrites and thioflavin-S-labeled amyloid plaques, we systematically analyzed the structural changes of dendrites associated with amyloid deposition in both a transgenic mouse model of AD (PSAPP) and in human postmortem brain. We found that in PSAPP mice, dendritic branches passing through or within 40 mum from amyloid deposits displayed various dendritic abnormalities such as loss of dendritic spines, shaft atrophy, bending, abrupt branch endings, varicosity formation, and sprouting. Similar structural alterations of dendrites were seen in postmortem human AD tissue, with spine loss as the most common abnormality in both PSAPP mice and human AD brains. These results demonstrate that fibrillar amyloid deposits and their surrounding microenvironment are toxic to dendrites and likely contribute to significant disruption of neuronal circuits in AD. PMID- 17413008 TI - Two-photon imaging of astrocytic Ca2+ signaling and the microvasculature in experimental mice models of Alzheimer's disease. AB - The sequence of events leading to neurodegeneration in Alzheimer's disease (AD) remains poorly understood. One prominent hypothesis is that neurovascular dysfunction contributes to both disease initiation and progression. Histologic analysis has supported this idea by demonstrating that vascular abnormalities are present early in the disease and most often perivascular amyloid deposits in the microvasculature. Two-photon in vivo imaging of mouse models of AD represents a unique approach to studying microvascular dysfunction in intact animals. We report here that a subpopulation of mice in early stages of AD (2-4 months) displays instability of vascular tone. Some, but not all animals exhibited oscillatory changes in arteriole diameter and poor vasodilation in response to sensory stimulation. An increased frequency of spontaneous astrocytic Ca(2+) increases was noted in animals with unstable vasculature. Because astrocytes recently have been shown to control local microcirculation and contribute to functional hyperemia, we suggest that abnormal astrocytic activity may contribute to vascular instability in AD and thereby to neuronal demise. PMID- 17413009 TI - Synaptic and mitochondrial morphometry provides structural correlates of successful brain aging. AB - Average synaptic size (S), synaptic numeric density (Nv) and surface density (Sv), average mitochondrial volume, mitochondrial numeric density, and mitochondrial volume density were measured by morphometry in the frontal (FC) and temporal (TC) cortex from adult and old monkeys (Macaca fascicularis). In relation to aging, Sv did not change, while Nv was significantly decreased in TC, but not in FC. S was significantly increased in FC and TC. No significant difference due to age was found with regard to mitochondrial ultrastructure. Considering the functional significance of the above parameters, their substantial age-related constancy suggests that they may reasonably represent structural correlates of successful brain aging. PMID- 17413010 TI - Impaired recognition memory and decreased prefrontal cortex spine density in aged female rats. AB - Aged F344 female rats (21 months) showed decreased performance, as compared to young rats (4 months), on an object recognition memory task. Golgi impregnation measured dendritic spine density of pyramidal neurons in the prefrontal cortex (layer II-III), a brain area important for recognition memory. Densities of spines in aged rats were 16% lower in tertiary, apical dendrites, but not significantly different in secondary basal dendrites. Concurrent measures of memory and spine density in the young and aged subjects show that age-related declines in recognition memory are associated with decreased cortical spine density. PMID- 17413011 TI - Alzheimer amyloid beta-peptide A-beta25-35 blocks adenylate cyclase-mediated forms of hippocampal long-term potentiation. AB - Progressive memory loss and deposition of amyloid beta (Abeta) peptides throughout cortical regions are hallmarks of Alzheimer's disease (AD). Several studies in mice and rats have shown that overexpression of amyloid precursor protein (APP) or pretreatment with Abeta peptide fragments results in the inhibition of hippocampal long-term potentiation (LTP) as well as impairments in learning and memory of hippocampal-dependent tasks. For these studies we have investigated the effects of the Abeta(25-35) peptide fragment on LTP induced by adenylate cyclase stimulation followed immediately by application of Mg(++)-free aCSF ("chemLTP"). Treatment of young adult slices with the Abeta(25-35) peptide had no significant effect on basal synaptic transmission in area CA1, but treatment with the peptide for 20 min before inducing chemLTP with isoproterenol (ISO; 1 microM) or forskolin (FSK;10 microM) + Mg(++)-free aCSF resulted in complete blockade of LTP. In contrast, normal ISO-chemLTP was observed after treatment with the control peptide Abeta(35-25). The ability of the Abeta(25-35) peptide fragment to block this and other forms of synaptic plasticity may help elucidate the mechanisms underlying hippocampal deficits observed in animal models of AD and/or AD individuals. PMID- 17413012 TI - Age-related changes in neuronal susceptibility to damage: comparison of the retinal ganglion cells of young and old mice before and after optic nerve crush. AB - To investigate whether or not the aging phenotype has increased vulnerability to axonal injury in vivo, we quantitated the loss of retinal ganglion cells (RGCs) after optic nerve crush. After crush, young animals lost 20% in 3 days and 50% of their RGCs in 7 days; however, old animals lost 40% in 3 days and 70% of their RGCs in 7 days. Our results showed that the time course in the loss of RGCs after crush in old mice is faster than that in young mice. Thus, old age increases susceptibility for the loss of RGCs following axonal damage. PMID- 17413013 TI - Top-down modulation and normal aging. AB - Normal aging is characterized by cognitive deficits that cross multiple domains and impair the ability of some older individuals to lead productive, high-quality lives. One of the primary goals of research in our laboratories is to study age related alterations in neural mechanisms that underlie a wide range of cognitive processes so that we may generate a unifying principle of cognitive aging. Top down modulation is the mechanism by which we enhance neural activity associated with relevant information and suppress activity for irrelevant information, thus establishing a foundation for both attention and memory processes. We use three converging technologies of human neurophysiology to study top-down modulation in aging: functional magnetic resonance imaging (fMRI), electroencephalography (EEG), and transcranial magnetic stimulation (TMS). Using these tools we have discovered that healthy older adults exhibit a selective inability to effectively suppress neural activity associated with distracting information and that this top-down suppression deficit is correlated with their memory impairment. We are now further characterizing the basis of these age-related alterations in top-down modulation and investigating interventions to remedy them. PMID- 17413014 TI - Brain aging and its modifiers: insights from in vivo neuromorphometry and susceptibility weighted imaging. AB - Aging is marked by individual differences and differential vulnerability of cognitive operations and their neural substrates. Cross-sectional studies of brain volume reveal greater age-related shrinkage of the prefrontal cortex (PFC) and the hippocampus than in the entorhinal and primary visual cortex. Longitudinal studies of regional brain shrinkage indicate that when individual differences are controlled, larger and broader shrinkage estimates are evident, with most polymodal cortices affected to the same extent. The mechanisms of age related shrinkage are unclear. Vascular risk factors may exacerbate brain aging and account for some of the observed declines as both the PFC and the hippocampus show elevated vulnerability to hypertension. MRI techniques that are sensitive to small vessels function, tissue oxygenation, and perfusion may be especially well suited to study brain aging and its vascular modifiers. We present an example of one such technique, susceptibility weighted imaging (SWI), that allows direct measurement of T2* values that reflect deoxy- to oxyhemoglobin fraction in blood vessels and iron deposits in cerebral tissue. The T2* shortening is associated with advanced age, but the effect is significantly stronger in the PFC and the hippocampus than the entorhinal and visual cortices. Moreover, T2* is shorter in hypertensive participants than in their matched normotensive counterparts, and the difference is especially prominent in the hippocampus, thus mirroring the findings of the neuromorphometric studies. Future research on brain aging would benefit from combining structural and metabolic techniques in a longitudinal design, as such studies will allow examination of leading-trailing effects of those factors. PMID- 17413015 TI - Linking brain imaging and genomics in the study of Alzheimer's disease and aging. AB - My colleagues and I have been using positron emission tomography (PET) and magnetic resonance imaging (MRI) to detect and track the brain changes associated with Alzheimer's disease (AD) and normal brain aging in cognitively normal persons with two copies, one copy, and no copies of the apolipoprotein E (APOE) epsilon4 allele, a common AD susceptibility gene. In this review article, I consider how brain imaging techniques could be used to evaluate putative AD prevention therapies in cognitively normal APOE epsilon4 carriers and putative age-modifying therapies in cognitively normal APOE epsilon4 noncarriers, how they could help investigate the individual and aggregate effects of putative AD risk modifiers, and how they could help guide the investigation of a molecular mechanism associated with AD vulnerability and normal neurological aging. I suggest how high-resolution genome-wide genetic and transcriptomic studies could further help in the scientific understanding of AD, aging, and other common and genetically complex phenotypes, such as variation in normal human memory performance, and in the discovery and evaluation of promising treatments for these phenotypes. Finally, I illustrate the push-pull relationship between brain imaging, genomics research, and other neuroscientific research in the study of AD and aging. PMID- 17413016 TI - Imaging and CSF studies in the preclinical diagnosis of Alzheimer's disease. AB - It is widely believed that the path to early and effective treatment for Alzheimer's disease (AD) requires the development of early diagnostic markers that are both sensitive and specific. To this aim, using longitudinal study designs, we and others have examined magnetic resonance imaging (MRI), 2-fluoro-2 deoxy-d-glucose-positron emission tomography (FDG/PET), and cerebrospinal fluid (CSF) biomarkers in cognitively normal elderly (NL) subjects and in patients with mild cognitive impairment (MCI). Such investigations have led to the often replicated findings that structural evidence of hippocampal atrophy as determined by MRI, as well as metabolic evidence from FDG-PET scan of hippocampal damage, predicts the conversion from MCI to AD. In this article we present a growing body of evidence of even earlier diagnosis. Brain pathology can be detected in NL subjects and used to predict future transition to MCI. This prediction is enabled by examinations revealing reduced glucose metabolism in the hippocampal formation (hippocampus and entorhinal cortex [EC]) as well as by the rate of medial temporal lobe atrophy as determined by MRI. However, neither regional atrophy nor glucose metabolism reductions are specific for AD. These measures provide secondary not primary evidence for AD. Consequently, we will also summarize recent efforts to improve the diagnostic specificity by combining imaging with CSF biomarkers and most recently by evaluating amyloid imaging using PET. We conclude that the combined use of conventional imaging, that is MRI or FDG-PET, with selected CSF biomarkers incrementally contributes to the early and specific diagnosis of AD. Moreover, selected combinations of imaging and CSF biomarkers measures are of importance in monitoring the course of AD and thus relevant to evaluating clinical trials. PMID- 17413017 TI - Functional MRI studies of associative encoding in normal aging, mild cognitive impairment, and Alzheimer's disease. AB - Functional magnetic resonance imaging (fMRI) is a noninvasive neuroimaging technique that can be used to study the neural correlates of complex cognitive processes, and the alterations in these processes that occur in the course of normal aging or superimposed neurodegenerative disease. Our studies have focused on the neural substrates of successful associative encoding, particularly of face name associations. We have found that the specific regions of the hippocampus and prefrontal cortices are critical for successful memory in both young and healthy older subjects. Our fMRI studies, as well as those of several other groups, have consistently demonstrated that, compared to cognitively intact older subjects, patients with clinical Alzheimer's disease (AD) have decreased fMRI activation in the hippocampus and related structures within the medial temporal lobe during the encoding of new memories. More recently, fMRI studies of subjects at risk for AD, by virtue of their genetics or evidence of mild cognitive impairment (MCI), have yielded variable results. Some of these studies, including our own, suggest that there may be a phase of paradoxically increased activation early in the course of prodromal AD. Further studies to validate fMRI in these populations are needed, particularly longitudinal studies to investigate the pattern of alterations in functional activity over the course of prodromal AD and the relationship to AD pathology. PMID- 17413018 TI - Quantitative EEG and electromagnetic brain imaging in aging and in the evolution of dementia. AB - Electroencephalographic (EEG) changes with normal aging have long been reported. Departures from age-expected changes have been observed in mild cognitive impairment and dementia, the magnitude of which correlates with the degree of cognitive impairment. Such abnormalities include increased delta and theta activity, decreased mean frequency, and changes in coherence. Similar findings have been reported using magnetoencephalography (MEG) at rest and during performance of mental tasks. Electrophysiological features have also been shown to be predictive of future decline in mild cognitive impairment (MCI) and Alzheimer's disease (AD). We have recently reported results from initial quantitative electroencephalography (QEEG) evaluations of normal elderly subjects (with only subjective reports of memory loss), predicting future cognitive decline or conversion to dementia, with high prediction accuracy (approximately 95%). In this report, source localization algorithms were used to identify the mathematically most probable underlying generators of abnormal features of the scalp-recorded EEG from these patients with differential outcomes. Using this QEEG method, abnormalities in brain regions identified in studies of AD using MEG, MRI, and positron emission tomography (PET) imaging were found in the premorbid recordings of those subjects who go on to decline or convert to dementia. PMID- 17413019 TI - [123I]5-IA-85380 SPECT imaging of beta2-nicotinic acetylcholine receptor availability in the aging human brain. AB - Human postmortem studies have reported decreases with age in high-affinity nicotine binding in brain. We have been investigating in vivo the availability of the beta(2)-containing nicotinic acetylcholine receptor (beta(2)-nAChR) in healthy nonsmokers (18-85 years of age) using [(123)I]5-IA-85380 SPECT imaging. Age and regional beta(2)-nAChR availability (V(T)(,)) have been observed to be inversely correlated in all brain regions analyzed, with decline ranging from 21% (cerebellum) to 36% (thalamus), or by up to 5% per decade of life. Preliminary results have confirmed postmortem reports of age-related decline in high-affinity nicotine binding with age and may elucidate the role of beta(2)-nAChRs in the cognitive decline associated with aging. PMID- 17413020 TI - Role of aerobic fitness and aging on cerebral white matter integrity. AB - Neuroimaging research suggests that cerebral white matter (WM) integrity, as reflected in fractional anisotropy (FA) via diffusion tensor imaging (DTI), is decreased in older adults, especially in the prefrontal regions of the brain. Behavioral investigations of cognitive functioning suggest that some aspects of cognition may be better preserved in older adults who possess higher levels of aerobic fitness. There are only a few studies, however, investigating potential mechanisms for the improvements in aerobic fitness. Our study suggests that greater aerobic fitness may be related to greater WM integrity in select brain regions. PMID- 17413021 TI - Age-related changes in nociceptive processing in the human brain. AB - Functional magnetic resonance imaging (fMRI) was used to compare cortical nociceptive responses to painful contact heat in healthy young (ages 22-30, n = 7) and older (ages 56-75, n = 7) subjects. Compared to young subjects, older subjects had significantly smaller pain-related fMRI responses in anterior insula (aINS) (P < 0.04), primary somatosensory cortex (S1) (P = 0.03), and supplementary motor area (P = 0.02). Gray matter volumes in S1 and aINS were significantly smaller for the older group (P = 0.02 and 0.0001, respectively), suggesting reduced processing capacity in these regions that might account for smaller pain-related fMRI responses. PMID- 17413022 TI - Magnetic resonance spectroscopy and environmental toxicant exposure. AB - The study of neurological impacts of toxicants has emphasized neuropsychological tests as important outcome variables. Direct assessment of neural substrates of environmental impacts could offer many advantages. I discuss our use of magnetic resonance spectroscopy (MRS) in the neurological assessment of adult lead poisoning of monozygotic twins as an example. Cognitive testing showed frontal lobe dysfunction in both twins, and more dramatic hippocampal dysfunction in the twin with higher lead exposure (JG). MRS showed lower N-acetylaspartate/creatine ratios in JG. The findings illustrate the potential utility of MRS in assessing impacts of not only lead, but other toxicants as well. PMID- 17413024 TI - Shifting paradigms in dementia: toward stratification of diagnosis and treatment using MRI. AB - Atrophy and cerebrovascular disease are the two most important magnetic resonance imaging (MRI) characteristics in the evaluation of dementia. On MRI, atrophy is the primary hallmark of neurodegenerative dementias including Alzheimer's disease (AD), while vascular dementia is characterized by the presence of ischemic vascular damage, such as territorial infarcts, lacunes, and white matter hyperintensities. Evidence is accumulating that vascular factors play an important role in the development of cognitive decline at old age and clinical AD. In the present article we present results of four recent MRI studies suggesting the additional involvement of small vessel disease in neurodegenerative disorders. Atrophy in the medial temporal lobe, as typically observed in AD, and small vessel disease often coincide. In terms of clinical significance, their effects may even be synergistic. The strict distinction between AD and vascular dementia is often artificial, as most patients suffer from both disorders to some extent. For the future, we see an important role for MRI in identifying those different compartments, regardless of clinical classification. Treatment could be directed by (and evaluated through) MRI patterns, rather than a diagnostic label. PMID- 17413025 TI - Imaging-guided microarray: isolating molecular profiles that dissociate Alzheimer's disease from normal aging. AB - Although both Alzheimer's disease (AD) and normal aging contribute to age-related hippocampal dysfunction, they are likely governed by separate molecular mechanisms. In principle, gene expression profiling can offer molecular clues about underlying mechanisms, but in practice techniques like microarray present unique analytic challenges when applied to disorders of the brain. Imaging-guided microarray is an approach designed to address these analytic challenges. Here, we will first review findings applying variants of functional magnetic resonance imaging (fMRI) to AD and normal aging, establishing the spatiotemporal profiles that dissociate one from the other. Then, we will review preliminary findings applying imaging-guided microarray to AD and normal aging, in an attempt to isolate molecular profiles that dissociate the two main causes of age-related hippocampal dysfunction. PMID- 17413023 TI - Tracking Alzheimer's disease. AB - Population-based brain mapping provides great insight into the trajectory of aging and dementia, as well as brain changes that normally occur over the human life span. We describe three novel brain mapping techniques, cortical thickness mapping, tensor-based morphometry (TBM), and hippocampal surface modeling, which offer enormous power for measuring disease progression in drug trials, and shed light on the neuroscience of brain degeneration in Alzheimer's disease (AD) and mild cognitive impairment (MCI). We report the first time-lapse maps of cortical atrophy spreading dynamically in the living brain, based on averaging data from populations of subjects with Alzheimer's disease and normal subjects imaged longitudinally with MRI. These dynamic sequences show a rapidly advancing wave of cortical atrophy sweeping from limbic and temporal cortices into higher-order association and ultimately primary sensorimotor areas, in a pattern that correlates with cognitive decline. A complementary technique, TBM, reveals the 3D profile of atrophic rates, at each point in the brain. A third technique, hippocampal surface modeling, plots the profile of shape alterations across the hippocampal surface. The three techniques provide moderate to highly automated analyses of images, have been validated on hundreds of scans, and are sensitive to clinically relevant changes in individual patients and groups undergoing different drug treatments. We compare time-lapse maps of AD, MCI, and other dementias, correlate these changes with cognition, and relate them to similar time-lapse maps of childhood development, schizophrenia, and HIV-associated brain degeneration. Strengths and weaknesses of these different imaging measures for basic neuroscience and drug trials are discussed. PMID- 17413026 TI - Fibrillar and oligomeric beta-amyloid as distinct local biomarkers for Alzheimer's disease. AB - Beta-amyloid is a key component of Alzheimer's disease (AD) pathology. Researchers in both academic and industry are actively pursuing the development of imaging tracers and techniques to noninvasively measure local levels of beta amyloid in the Alzheimer's brain. This presentation summarizes recent data and discusses the opportunities and challenges of imaging plaques containing fibrillar beta-amyloid for the early diagnosis and therapeutic monitoring of amyloid targeted therapies. Further, the value and feasibility of measuring the recently described soluble oligomeric form of beta-amyloid as an alternative noninvasive biomarker is also discussed. PMID- 17413027 TI - Diffusion tensor imaging of normal appearing white matter and its correlation with cognitive functioning in mild cognitive impairment and Alzheimer's disease. AB - Diffusion tensor imaging (DTI) was used to examine the microstructural integrity of normal appearing white matter (NAWM) in subjects with mild cognitive impairment (MCI) and Alzheimer's disease (AD). Significant frontal, temporal, and parietal white matter diffusion tensor changes were demonstrated in MCI and AD compared with normal controls. These changes were correlated with cognitive functioning, and are consistent with a hypothesized loss of axonal processes in affected regions. PMID- 17413028 TI - Enhanced ryanodine-mediated calcium release in mutant PS1-expressing Alzheimer's mouse models. AB - Intracellular Ca(2+) signaling involves Ca(2+) liberation through both inositol triphosphate and ryanodine receptors (IP(3)R and RyR). However, little is known of the functional interactions between these Ca(2+) sources in either neuronal physiology, or during Ca(2+) disruptions associated with Alzheimer's disease (AD). By the use of whole-cell recordings and 2-photon Ca(2+) imaging in cortical slices we distinguished between IP(3)R- and RyR-mediated Ca(2+) components in nontransgenic (non-Tg) and AD mouse models and demonstrate powerful signaling interactions between these channels. Ca(2+)-induced Ca(2+) release (CICR) through RyR contributed modestly to Ca(2+) signals evoked by photoreleased IP(3) in cortical neurons from non-Tg mice. In contrast, the exaggerated signals in 3xTg AD and PS1(KI) mice resulted primarily from enhanced CICR through RyR, rather than through IP(3)R, and were associated with increased RyR expression levels. Moreover, membrane hyperpolarizations evoked by IP(3) in neurons from AD mouse models were even greater than expected simply from the exaggerated Ca(2+) signals, pointing to an increased coupling efficiency between cytosolic [Ca(2+)] and K(+) channel regulation. Our results highlight the critical roles of RyR mediated Ca(2+) signaling in both neuronal physiology and pathophysiology, and point to presenilin-linked disruptions in RyR signaling as an important genetic factor in AD. PMID- 17413029 TI - Prospects for prediction: ethics analysis of neuroimaging in Alzheimer's disease. AB - This article focuses on the prospects and ethics of using neuroimaging to predict Alzheimer's disease (AD). It is motivated by consideration of the historical roles of science in medicine and society, and considerations specifically contemporary of capabilities in imaging and aging, and the benefits and hope they bring. A general consensus is that combinations of imaging methods will ultimately be most fruitful in predicting disease. Their roll-out into translational practice will not be free of complexity, however, as culture and values differ in terms of what defines benefit and risk, who will benefit and who is at risk, what methods must be in place to assure the maximum safety, comfort, and protection of subjects and patients, and educational and policy needs. Proactive planning for the ethical and societal implications of predicting diseases of the aging brain is critical and will benefit all stakeholders researchers, patients and families, health care providers, and policy makers. PMID- 17413030 TI - Deficiency in Nrf2-GSH signaling impairs type II cell growth and enhances sensitivity to oxidants. AB - Redox imbalance has been implicated in the pathogenesis of many acute and chronic lung diseases. The b-Zip transcription factor Nrf2 acts via an antioxidant/electrophilic response element to regulate antioxidants and maintain cellular redox homeostasis. Our previous studies have shown that Nrf2-deficient mice (Nrf2(-/-)) show reduced pulmonary expression of several antioxidant enzymes, which renders them highly susceptible to hyperoxia-induced lung injury. To better understand the physiologic significance of Nrf2-induced redox signaling, we have used primary cells isolated from the lungs of Nrf2(+/+) and Nrf2(-/-) mice. Our studies were focused on type II cells because these cells are constantly exposed to the oxidant environment and play key roles in host defense, injury, and repair processes. Using this system, we now report that an Nrf2 deficiency leads to defects in type II cell proliferation and greatly enhances the cells' sensitivity to oxidant-induced cell death. These defects were closely associated with high levels of reactive oxygen species (ROS) and redox imbalance in Nrf2(-/-) cells. Glutathione (GSH) supplementation rescued these phenotypic defects associated with the Nrf2 deficiency. Intriguingly, although the antioxidant N-acetyl-cysteine drastically squelched ROS levels, it was unable to counteract growth arrest in Nrf2(-/-) cells. Moreover, despite their elevated levels of ROS, Nrf2(-/-) type II cells were viable and, like their wild-type counterparts, exhibited normal differentiation characteristics. Our data suggest that dysfunctional Nrf2-regulated GSH-induced signaling is associated with deregulation of type II cell proliferation, which contributes to abnormal injury and repair and leads to respiratory impairment. PMID- 17413031 TI - Transcriptional profiling of mucociliary differentiation in human airway epithelial cells. AB - When cultured at an air-liquid interface (ALI) in the appropriate medium, primary human airway epithelial cells form a polarized, pseudostratified epithelium composed of ciliated and mucus-secreting cells. This culture system provides a useful tool for the in vitro study of airway epithelial biology and differentiation. We have performed microarray analysis on ALI cultures of human bronchial epithelial cells (HBECs) grown over a 28-d period to identify genes involved in mucociliary differentiation. We identified over 2,000 genes that displayed statistically significant 2-fold or greater changes in expression during the time course. Of the genes showing the largest increases, many are involved in processes associated with airway epithelial biology, such as cell adhesion, immunity, transport, and cilia formation; however, many novel genes were also identified. We compared our results with data from proteomic analyses of the ciliary axoneme and identified candidate genes that may have roles in cilia formation or function. Gene networks were generated using Ingenuity Pathways Analysis (Ingenuity Systems, Redwood City, CA) to identify signaling pathways involved in mucociliary cell differentiation or function. Networks containing genes involved in TGF-beta, WNT/beta-catenin, and epidermal growth factor receptor (EGFR) pathways were identified, suggesting potential roles for these families in airway epithelia. Microarray results were validated by real time RT-PCR for a number of representative genes. This work has provided extensive information about gene expression changes during differentiation of airway epithelial cells, and will be a useful resource for researchers interested in respiratory function, pathology, and toxicology. PMID- 17413032 TI - G1 phase cell cycle arrest induced by SARS-CoV 3a protein via the cyclin D3/pRb pathway. AB - SARS-CoV 3a is a structural protein, mainly localizing to Golgi apparatus and co localizing with SARS-CoV M in co-transfected cells. Here we observed that transient expression of 3a inhibited cell growth and prevented 5 bromodeoxyuridine incorporation, suggesting that 3a deregulated cell cycle progression. Cell cycle analysis demonstrated that 3a expression was associated with blockage of cell cycle progression at G1 phase in HEK 293, COS-7, and Vero cells 24-60 h after transfection. Mutation analysis of 3a revealed that C terminal region (176 aa approximately 274 aa), including a potential calcium ATPase motif, was essential for induction of cell cycle arrest. Topological analysis showed that 3a predominantly located in Golgi apparatus, with its N terminus residing in the lumen (Nlum) and C-terminus in the cytosol (Ccyt). Analyzing the cellular proteins involving in regulation of cell cycle progression, we demonstrated that 3a expression was correlated with a significant reduction of cyclin D3 level and phosphorylation of retinoblastoma (Rb) protein at Ser-795 and Ser-809/811, not with the expression of cyclin D1, D2, cdk4, and cdk6 in 293 cells. Increases in p53 phosphorylation on Ser-15 were observed in both SARS-CoV M and 3a transfected cells, suggesting that it might not correlate with the 3a-induced G0/G1 phase arrest. The reduction of cyclin D3 level and phosphorylation of Rb were further confirmed in SARS-CoV infected Vero cells. These results indicate that SARS-CoV 3a protein, through limiting the expression of cyclin D3, may inhibit Rb phosphorylation, which in turn leads to a block in the G1 phase of the cell cycle and an inhibition of cell proliferation. PMID- 17413033 TI - Expression of heme oxygenase-1 in human vascular cells is regulated by peroxisome proliferator-activated receptors. AB - OBJECTIVE: Activation of peroxisome proliferator-activated receptors (PPARs) by lipid-lowering fibrates and insulin-sensitizing thiazolidinediones inhibits vascular inflammation, atherosclerosis, and restenosis. Here we investigate if the vasculoprotective and anti-inflammatory enzyme heme oxygenase-1 (HO-1) is regulated by PPAR ligands in vascular cells. METHODS AND RESULTS: We show that treatment of human vascular endothelial and smooth muscle cells with PPAR ligands leads to expression of HO-1. Analysis of the human HO-1 promoter in transient transfection experiments together with mutational analysis and gel shift assays revealed a direct transcriptional regulation of HO-1 by PPARalpha and PPARgamma via 2 PPAR responsive elements. We demonstrate that a clinically relevant polymorphism within the HO-1 promoter critically influences its transcriptional activation by both PPAR isoforms. Moreover, inhibition of HO-1 enzymatic activity reversed PPAR ligand-mediated inhibition of cell proliferation and expression of cyclooxygenase-2 in vascular smooth muscle cells. CONCLUSION: We demonstrate that HO-1 expression is transcriptionally regulated by PPARalpha and PPARgamma, indicating a mechanism of anti-inflammatory and antiproliferative action of PPAR ligands via upregulation of HO-1. Identification of HO-1 as a target gene for PPARs provides new strategies for therapy of cardiovascular diseases and a rationale for the use of PPAR ligands in the treatment of other chronic inflammatory diseases. PMID- 17413034 TI - TNF-alpha contributes to endothelial dysfunction by upregulating arginase in ischemia/reperfusion injury. AB - BACKGROUND: We tested whether tumor necrosis factor (TNF)-alpha increases arginase expression in endothelial cells as one of the primary mechanisms by which this inflammatory cytokine compromises endothelial function during ischemia reperfusion (I/R) injury. METHODS AND RESULTS: Mouse hearts were subjected to 30 minutes of global ischemia followed by 90 minutes of reperfusion and their vasoactivity before and after I/R was examined in wild-type (WT), tumor necrosis factor knockout (TNF-/-), and TNF 1.6 (TNF++/++) mice. In WT mice, dilation to the endothelium-dependent vasodilator ACh was blunted in I/R compared with sham control. L-arginine or arginase inhibitor NOHA restored NO-mediated coronary arteriolar dilation in WT I/R mice. O2(-) production was reduced by eNOS inhibitor, L-NAME, or NOHA in WT I/R mice. In TNF-/- mice, I/R did not alter Ach induced vasodilation and O2(-) production compared with sham mice. The increase in arginase expression that occurs during I/R in WT mice was absent in TNF-/- mice. Arginase expression was confined largely to the endothelium and independent of inflammatory cell invasion. Arginase activity was markedly lower in TNF-/-, but higher in WT I/R than that in WT sham mice. CONCLUSIONS: Our data demonstrate TNF-alpha upregulates expression of arginase in endothelial cells, which leads to O2(-) production then induces endothelial dysfunction in I/R injury. PMID- 17413035 TI - (6R)-5,6,7,8-tetrahydro-L-biopterin and its stereoisomer prevent ischemia reperfusion injury in human forearm. AB - OBJECTIVE: 6R-5,6,7,8-tetrahydro-L-biopterin (6R-BH4) is a cofactor for endothelial nitric oxide synthase but also has antioxidant properties. Its stereo isomer 6S-5,6,7,8-tetrahydro-L-biopterin (6S-BH4) and structurally similar pterin 6R,S-5,6,7,8-tetrahydro-D-neopterin (NH4) are also antioxidants but have no cofactor function. When endothelial nitric oxide synthase is 6R-BH4-deplete, it synthesizes superoxide rather than nitric oxide. Reduced nitric oxide bioavailability by interaction with reactive oxygen species is implicated in endothelial dysfunction (ED). 6R-BH4 corrects ED in animal models of ischemia reperfusion injury (IRI) and in patients with cardiovascular risks. It is uncertain whether the effect of exogenous 6R-BH4 on ED is through its cofactor or antioxidant action. METHODS AND RESULTS: In healthy volunteers, forearm blood flow was measured by venous occlusion plethysmography during intra-arterial infusion of the endothelium-dependent vasodilator acetylcholine, or the endothelium-independent vasodilator glyceryl trinitrate, before and after IRI. IRI reduced plasma total antioxidant status (P=0.03) and impaired vasodilatation to acetylcholine (P=0.01), but not to glyceryl trinitrate (P=0.3). Intra-arterial infusion of 6R-BH4, 6S-BH4 and NH4 at approximately equimolar concentrations prevented IRI. CONCLUSION: IRI causes ED associated with increased oxidative stress that is prevented by 6R-BH4, 6S-BH4, and NH4, an effect mediated perhaps by an antioxidant rather than cofactor function. Regardless of mechanism, 6R-BH4, 6S-BH4, or NH4 may reduce tissue injury during clinical IRI syndromes. PMID- 17413036 TI - Low levels of Nogo-B in human carotid atherosclerotic plaques are associated with an atheromatous phenotype, restenosis, and stenosis severity. AB - OBJECTIVE: Reticulon-4/Nogo (Nogo-B) protects mouse arteries from lumen loss by reducing smooth muscle cell (SMC) migration and intimal thickening. Our goal was to determine plaque and circulating levels of Nogo-B in atherosclerotic and control subjects. Therefore, we studied the relationships between local Nogo-B, plaque characteristics, and clinical data in patients undergoing carotid endarterectomy. METHODS AND RESULTS: Western blot analysis showed that endarterectomy specimens from the femoral (n=19) and carotid arteries (n=145) contained significantly less Nogo-B than nonatherosclerotic mammary arteries (n=8; P<0.003) and aortas (n=15; P=0.03). Immunohistochemistry revealed that in atherosclerotic lesions, Nogo-B was expressed by macrophage/foam cells, SMC rich, and neo-vascularized areas. Atheromatous plaques (>40% fat content) showed a significant reduction in Nogo-B expression (P=0.002). Nogo-B expression levels were significantly lower in patients with more than 90% of carotid stenosis (P=0.04) or restenotic lesions after prior carotid intervention (duplex; P=0.01). In contrast, plasmatic levels of Nogo-B (soluble Nogo-B) did not differ between atherosclerotic subjects (n=68) and risk-factor matched controls (n=63; P=0.5). CONCLUSION: Our findings suggest that local reduction of Nogo-B in atherosclerotic tissue might contribute to plaque formation and/or instability triggering luminal narrowing. In contrast, plasma Nogo-B levels are not associated with clinically manifested atherosclerotic disease. PMID- 17413037 TI - Haplotypes of IL1B, IL1RN, IL1R1, and IL1R2 and the risk of venous thrombosis. AB - OBJECTIVE: It has been suggested that the overall effect of the major proinflammatory cytokine interleukin-1 (IL-1) on coagulation and fibrinolysis is prothrombotic. The aim of this study was to investigate whether common variations in IL1B, IL1RN, IL1R1, and IL1R2 influence the risk of venous thrombosis. METHODS AND RESULTS: In a case-control study on the causes of deep venous thrombosis, the Leiden Thrombophilia Study (LETS), we genotyped 18 single nucleotide polymorphisms (SNPs) in IL1B, IL1RN, IL1R1, and IL1R2, enabling us to tag a total of 25 haplotype groups. Overall testing of the haplotype frequency distribution in patients and controls indicated that a recessive effect was present in IL1RN (P=0.031). Subsequently the risk of venous thrombosis was calculated for each haplotype of IL1RN. Increased thrombotic risk was found for homozygous carriers of haplotype 5 (H5, tagged by SNP 13888T/G, rs2232354) of IL1RN (Odds ratio=3.9; 95% confidence interval: 1.6 to 9.7; P=0.002). No risk was associated with haplotype 3 of IL1RN, which contains the frequently examined allele 2 variant of the intron 2 VNTR. CONCLUSIONS: We found that IL1RN-H5H5 carriership increases the risk of venous thrombosis. PMID- 17413038 TI - A role for the aryl hydrocarbon receptor in regulation of ischemia-induced angiogenesis. AB - OBJECTIVE: The aryl hydrocarbon receptor (AHR) is a transcription factor that binds to DNA as a heterodimer with the AHR nuclear translocator (ARNT) after interaction with ligands such as polycyclic and halogenated aromatic hydrocarbons found in tobacco smoke and the environment. We have investigated the interaction between AHR and hypoxia signaling pathways in regulation of angiogenesis with the use of a surgical model of ischemia. METHODS AND RESULTS: Ischemia was induced by femoral artery occlusion in wild-type and AHR-null mice. Ischemia-induced angiogenesis was markedly enhanced in AHR-null mice compared with that in wild type animals. Ischemia-induced upregulation of the expression of hypoxia inducible factor-1alpha (HIF-1alpha) and ARNT as well as that of target genes for these transcription factors, such as that for vascular endothelial growth factor (VEGF), were also enhanced in AHR-null mice. Furthermore, the DNA binding activity of the HIF-1alpha-ARNT complex as well as the association of HIF-1alpha and ARNT with the VEGF gene promoter were increased by ischemia to a greater extent in AHR-null mice than in wild-type animals. CONCLUSIONS: Ablation of AHR resulted in enhancement of ischemia-induced angiogenesis. This effect was likely attributable in part to the associated enhancement of ischemia-induced VEGF expression, which in turn may be caused by an increased abundance and activity of the HIF-1alpha-ARNT heterodimer. PMID- 17413039 TI - Modulation of tissue factor-factor VIIa signaling by lipid rafts and caveolae. AB - OBJECTIVE: Coagulation factor VIIa (VIIa) binding to its cellular receptor, tissue factor (TF), not only initiates the coagulation cascade but also induces cell signaling by activating G-protein coupled protease-activated receptors. The objective of the present study is to investigate the role of lipid rafts and caveolae in modulating TF-VIIa signaling and coagulant functions. METHODS AND RESULTS: TF-VIIa coagulant function was measured in factor X activation assay and the signaling function was evaluated in phosphoinositide hydrolysis and IL-8 gene induction. Buoyant density gradient centrifugation and immunofluorescence confocal microscopy were used to determine cellular localization of TF and protease-activated receptor 2. The data show that a substantial fraction of TF and protease-activated receptor 2 resides in lipid rafts/caveolae, and disruption of lipid rafts by cholesterol depletion or modification reduced TF-VIIa-induced cell signaling. Disruption of caveolae with caveolin-1 silencing had no effect on the TF-VIIa coagulant activity but inhibited the TF-VIIa-induced cell signaling. CONCLUSION: Overall our data show that lipid raft/caveolae play a selective role in modulating the TF-VIIa signaling function without affecting the TF-VIIa coagulant activity. PMID- 17413040 TI - Plasmin triggers cytokine induction in human monocyte-derived macrophages. AB - OBJECTIVE: Fibrinolytic activity is upregulated in atherosclerotic lesions, yet little is known about the role of plasmin in macrophage function. We postulated a direct effect of plasmin on human monocyte-derived macrophages. METHODS AND RESULTS: Plasmin activates macrophages via the annexin A2 heterotetramer composed of annexin A2 and S100A10 with subsequent stimulation of Janus kinase JAK1/TYK2 signaling. JAK1/TYK2 leads to STAT3 activation, Akt-dependent NF-kappaB activation, and phosphorylation of extracellular signal-regulated kinase 1/2 and mitogen-activated kinase p38. These signaling pathways trigger nuclear translocation of STAT3 and p65 transcription factors and the induction of the proinflammatory cytokines tumor necrosis factor-alpha and IL-6. Inhibitors of JAK, p38, and NF-kappaB revealed that these signaling pathways are indispensable for the plasmin-mediated tumor necrosis factor-alpha and IL-6 induction. By contrast, the extracellular signal-regulated kinase 1/2 activation is essential only for the IL-6 expression. The activation clearly depends on the proteolytic activity of plasmin, which cleaves the A2 subunit of the annexin A2 heterotetramer. Downregulation of each of the receptor subunits by antisense oligodeoxynucleotides abolished the plasmin-induced expression of proinflammatory cytokines stressing the crucial role the annexin A2 heterotetramer. CONCLUSIONS: Plasmin generated at sites of inflammation such as atherosclerotic lesions will trigger cytokine expression in human macrophages. PMID- 17413041 TI - Interaction of alpha9beta1 integrin with thrombospondin-1 promotes angiogenesis. AB - Thrombospondin-1 is a multifunctional protein interacting with several cell surface receptors including integrins. We found that it is a ligand for alpha9beta1 integrin, and has an integrin binding site within its N-terminal domain (NoC1). Interaction of thrombospondin-1 and its recombinant NoC1 domain with alpha9beta1 integrin was confirmed in ELISA and cell adhesion assays. Binding of NoC1 to cells expressing alpha9beta1 integrin activated signaling proteins such as Erk1/2 and paxillin. Blocking of this integrin by monoclonal antibody and the met-leu-asp-disintegrin inhibited dermal human microvascular endothelial cell proliferation and NoC1-induced migration of these cells. Immunohistochemical studies revealed that alpha9beta1 is expressed on microvascular endothelium in several organs including skin, lung, heart and brain. NoC1 induced neovascularization in an experimental quail chorioallantoic membrane system and Matrigel plug formation assay in mice. This proangiogenic activity of NoC1 in vivo was inhibited by alpha9beta1 inhibitors. In summary, our results revealed that alpha9beta1 integrin expressed on microvascular endothelial cells interacts with thrombospondin-1, and this interaction is involved in modulation of angiogenesis. PMID- 17413042 TI - CREB mediates UTP-directed arterial smooth muscle cell migration and expression of the chemotactic protein osteopontin via its interaction with activator protein 1 sites. AB - The transcription factor cAMP responsive element-binding protein (CREB) has been found to be involved in arterial smooth muscle cell (SMC) migration. We previously demonstrated that osteopontin (OPN) expression is a key step for UTP mediated migration of arterial SMCs and that activator protein (AP)-1, nuclear factor kappaB, and upstream stimulatory transcription factors are involved in this OPN expression. The present study aims to determine the role of CREB in UTP induced migration and OPN expression in cultured SMCs. We found that CREB is activated by UTP via extracellular signal-regulated kinase 1/2 and p38 mitogen activated protein kinase pathways but not by protein kinase A. Both overexpression of a dominant negative CREB and CREB small interfering RNA treatment suppressed UTP-induced OPN expression and SMC migration. Gel-shift and chromatin immunoprecipitation assays revealed that CREB binds 2 AP-1 sites (-1870 and -76) and a cAMP responsive element-like site (-1403) on the OPN promoter. Mutations of these sites showed that only the 2 AP-1 sites were required for UTP induced OPN expression. Moreover, gel-supershift and sequential chromatin immunoprecipitation assays suggested that CREB was associated with c-Fos on the AP-1 sites of the OPN promoter. These results demonstrate that CREB participates in the induction of UTP-activated OPN expression via its binding to 2 AP-1 sites and is thus involved in UTP-mediated SMC migration. PMID- 17413043 TI - Increased ventricular preload is compensated by myocyte proliferation in normal and hypoplastic fetal chick left ventricle. AB - Hemodynamics influence cardiac development, and alterations in blood flow may lead to impaired cardiac growth and malformations. The developing myocardium adapts to augmented workload by increasing cell number (hyperplasia). The aim of this study was to determine the influence of alterations in ventricular preload on fetal myocyte proliferation by manipulation of intracardiac shunting at the atrial level. We hypothesized that partial clipping of the right atrial appendage would increase the blood flow to the left ventricle and, in turn, lead to an increase in chamber volume and myocardial mass based on myocyte proliferation. Using an ex ovo culture setup, we performed partial right atrial clipping on embryonic day 8 chick embryos. Ultrasound imaging was performed before and after the surgery to assess the changes in left ventricular volume. Sampling after 24 hours was preceded by 2 hour of pulse-labeling with 5-bromodeoxyuridine. Ultrasound imaging showed that partial right atrial clipping led to a significant increase in left ventricular end-diastolic volume, demonstrating increased blood flow and preload. Anti-5-bromodeoxyuridine immunolabeling revealed a significant increase in myocyte proliferation in the left ventricle and atrium. No significant changes were found in the right heart structures. Increased left ventricular myocyte proliferation and myocardial mass after right atrial clipping was also observed in embryos with experimental left ventricular hypoplasia. These results demonstrate the ability of fetal myocardium to respond to increased preload by myocyte hyperplasia and support the rationale for prenatal surgical interventions in certain cases of congenital heart disease such as hypoplastic left heart syndrome. PMID- 17413044 TI - Beta-catenin downregulation is required for adaptive cardiac remodeling. AB - The armadillo-related protein beta-catenin has multiple functions in cardiac tissue homeostasis: stabilization of beta-catenin has been implicated in adult cardiac hypertrophy, and downregulation initiates heart formation in embryogenesis. The protein is also part of the cadherin/catenin complex at the cell membrane, where depletion might result in disturbed cell-cell interaction similar to N-cadherin knockout models. Here, we analyzed the in vivo role of beta catenin in adult cardiac hypertrophy initiated by angiotensin II (Ang II). The cardiac-specific mifepristone-inducible alphaMHC-CrePR1 transgene was used to induce beta-catenin depletion (loxP-flanked exons 3 to 6, beta-cat(Deltaex3-6) mice) or stabilization (loxP-flanked exon 3, beta-cat(Deltaex3) mice). Levels of beta-catenin were altered both in membrane and nuclear extracts. Analysis of the beta-catenin target genes Axin2 and Tcf-4 confirmed increased beta-catenin dependent transcription in beta-catenin stabilized mice. In both models, transgenic mice were viable and healthy at age 6 months. beta-Catenin appeared dispensable for cell membrane function. Ang II infusion induced cardiac hypertrophy both in wild-type mice and in mice with beta-catenin depletion. In contrast, mice with stabilized beta-catenin had decreased cross-sectional area at baseline and an abrogated hypertrophic response to Ang II infusion. Stabilizing beta-catenin led to impaired fractional shortening compared with control littermates after Ang II stimulation. This functional deterioration was associated with altered expression of the T-box proteins Tbx5 and Tbx20 at baseline and after Ang II stimulation. In addition, atrophy-related protein IGFBP5 was upregulated in beta-catenin-stabilized mice. These data suggest that beta-catenin downregulation is required for adaptive cardiac hypertrophy. PMID- 17413045 TI - Regulation of L-type calcium channel and delayed rectifier potassium channel activity by p21-activated kinase-1 in guinea pig sinoatrial node pacemaker cells. AB - Phosphorylation of ion channels plays an important role in the regulation of cardiac function, but signaling mechanisms controlling dephosphorylation are not well understood. We have tested the hypothesis that p(21)-activated kinase-1 (Pak1), a serine-threonine protein kinase regulated by Ras-related small G proteins, regulates sinoatrial node (SAN) ion channel activity through a mechanism involving protein phosphatase 2A. We report a novel role of Pak1 mediated signaling in attenuating isoproterenol-induced enhancement of L-type Ca(2+) current (I(CaL)) and delayed rectifier potassium current (I(K)) in guinea pig SAN pacemaker cells. We demonstrate that in guinea pig SAN: (1) there is abundant expression of endogenous Pak1 in pacemaker cells; (2) expression of constitutively active Pak1 depresses isoproterenol-induced upregulation of I(CaL) and I(K); (3) inhibition of protein phosphatase 2A increases the enhancement of I(K) and I(CaL) by isoproterenol in Ad-Pak1-infected cells; (4) protein phosphatase 2A coimmunoprecipitates with endogenous Pak1 in SAN tissue; and (5) expression of constitutively active Pak1 suppresses the chronotropic action of isoproterenol on pacemaker activity of intact SAN preparations. In conclusion, our data demonstrate that a Pak1 signaling pathway exists in cardiac pacemaker cells and that this novel pathway plays a role in the regulation of ion channel activity. PMID- 17413046 TI - The 2006 Thomas Willis lecture: the adventures of a translational researcher in stroke and migraine. PMID- 17413047 TI - Effects of acute pravastatin treatment on intensity of rescue therapy, length of inpatient stay, and 6-month outcome in patients after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: We have previously demonstrated that acute pravastatin therapy after aneurysmal subarachnoid hemorrhage ameliorates vasospasm-related delayed ischemic deficits. This study assesses the effects of pravastatin on the frequency and intensity for rescue therapy, length of inpatient stay, and long term outcome at 6 months. METHODS: Eighty aneurysmal subarachnoid hemorrhage patients (age 18 to 84 years, onset 1.8+/-1.3 days) were randomized to receive daily oral pravastatin (40 mg) or placebo for up to 14 days. Clinical events were recorded during the trial. Six-month outcome was assessed using the Short Form 36 and the modified Rankin Scale. RESULTS: Although no significant difference in the outcome at discharge was found between the trial groups, multivariate analysis showed pravastatin therapy reduced unfavorable outcome by 73% (P=0.041). The benefit persisted at 6 months (P=0.063) and was notable in the physical (P<0.001) and psychosocial (P<0.001) aspects measured using Short Form 36. Furthermore, the acute pravastatin therapy reduced the requirement for triple-H therapy (hypertensive, hypervolemic, hemodilution; P=0.045) and mortality related to vasospasm (P=0.02) and sepsis (P=0.001); no significant difference was found in the length of inpatient stay between the trial groups. CONCLUSIONS: This trial demonstrates that acute statin treatment reduces traditional rescue therapy for vasospasm after aneurysmal subarachnoid hemorrhage. Improvement in early outcome has proved robust at 6 months, particularly in relation to physical and psychosocial (Short Form 36) outcome. PMID- 17413048 TI - Hyperoxic reperfusion after global ischemia decreases hippocampal energy metabolism. AB - BACKGROUND AND PURPOSE: Previous reports indicate that compared with normoxia, 100% ventilatory O(2) during early reperfusion after global cerebral ischemia decreases hippocampal pyruvate dehydrogenase activity and increases neuronal death. However, current standards of care after cardiac arrest encourage the use of 100% O(2) during resuscitation and for an undefined period thereafter. Using a clinically relevant canine cardiac arrest model, in this study we tested the hypothesis that hyperoxic reperfusion decreases hippocampal glucose metabolism and glutamate synthesis. METHODS: After 10 minutes of cardiac arrest, animals were resuscitated and ventilated for 1 hour with 100% O(2) (hyperoxic) or 21% to 30% O(2) (normoxic). At 30 minutes reperfusion, [1-(13)C]glucose was infused, and at 2 hours, brains were rapidly removed and frozen. Extracted metabolites were analyzed by (13)C nuclear magnetic resonance spectroscopy. RESULTS: Compared with nonischemic controls, the hippocampi from hyperoxic animals had elevated levels of unmetabolized (13)C-glucose and decreased incorporation of (13)C into all isotope isomers of glutamate. These findings indicate impaired neuronal metabolism via the pyruvate dehydrogenase pathway for carbon entry into the tricarboxylic acid cycle and impaired glucose metabolism via the astrocytic pyruvate carboxylase pathway. No differences were observed in the cortex, indicating that the hippocampus is more vulnerable to metabolic changes induced by hyperoxic reperfusion. CONCLUSIONS: These results represent the first direct evidence that hyperoxia after cardiac arrest impairs hippocampal oxidative energy metabolism in the brain and challenge the rationale for using excessively high resuscitative ventilatory O(2). PMID- 17413049 TI - Asymmetric dynamic cerebral autoregulatory response to cyclic stimuli. AB - BACKGROUND AND PURPOSE: Dynamic cerebral autoregulation has been shown to be fast and effective, but it is not well known if the mechanism is symmetric, that is to say, it acts with equal compensatory action to upward as compared with downward abrupt changes in arterial blood pressure (ABP). METHODS: Fourteen patients with head injuries and 10 normal subjects had bilateral transcranial Doppler and continuous ABP recording. Cyclic ABP stimuli were generated by large thigh cuffs, which were rapidly inflated above systolic pressure for 15 seconds alternating with 15 seconds of deflation. At least 8 such cycles were ensemble-averaged and the dynamic autoregulatory gain (AG(up) and AG(dn)) was estimated separately for upward and downward changes in ABP. The results were compared with the autoregulation index using conventional leg cuff releases. RESULTS: In normal subjects, AG(dn) was 0.74+/-0.18 and AG(up) was 0.77+/-0.17 (mean+/-SD); the difference was insignificant. The correlation between AG(dn) and AG(up), however, was weak (r=0.24). In the patients with head injury, AG(dn) was 0.30+/-0.21 and AG(up) was 1.27+/-0.76, the difference being highly significant (P<0.001). There was a negative relationship between AG(dn) and AG(up) (r=-0.33). Autoregulation index correlated well with AG(dn) (r=0.79) and weakly negatively with AG(up) (r= 0.47). CONCLUSIONS: A strongly asymmetric dynamic response of the cerebral autoregulation was seen the majority of patients with head injury. It might also have been present, albeit to a lesser degree, in the normal subjects. The findings suggest that nonlinear effects may be present in the operation of the cerebral autoregulation mechanism. PMID- 17413050 TI - Increased risk of childhood asthma from antibiotic use in early life. AB - BACKGROUND: To address the major methodological issues of reverse causation and selection bias in epidemiologic studies of antibiotic use in early life and the development of asthma, we undertook a cohort study of this association in a complete population of children. METHODS: Using the health-care and prescription databases of Manitoba, Canada, this longitudinal study assessed the association between antibiotic prescription use during the first year of life and asthma at age 7 years in a 1995 birth cohort of 13,116 children. RESULTS: Independent of well-known asthma risk factors, asthma was significantly more likely to develop in children who had received antibiotics in the first year of life at age 7 years. The association with asthma was observed for antibiotic use in non respiratory tract infections (adjusted odds ratio [OR], 1.86; 95% confidence interval [CI], 1.02 to 3.37). The risk of asthma was highest in children receiving more than four courses of antibiotics (adjusted OR, 1.46; 95% CI, 1.14 to 1.88), especially among rural children, and in the absence of maternal asthma or a dog in the birth year. Broad-spectrum (BS) cephalosporin use was more common in these subpopulations of children. CONCLUSIONS: Antibiotic use in early life was associated with the development of childhood asthma, a risk that may be reduced by avoiding the use of BS cephalosporins. PMID- 17413051 TI - Postobstructive pulmonary edema: a case for hydrostatic mechanisms. AB - BACKGROUND: Postobstructive pulmonary edema is a well-recognized complication of upper airway obstruction. The mechanisms of edema formation are unclear and may be due to increased hydrostatic forces generated by high negative inspiratory pressure or by increased permeability of the alveolar capillary membrane. Measurement of the edema fluid/plasma protein ratio and the rate of net alveolar fluid clearance are two well-validated methods for classifying the underlying mechanism of edema formation. The goal of the current study was to investigate the mechanisms of pulmonary edema formation in patients with postobstructive pulmonary edema by serial sampling of undiluted pulmonary edema fluid. METHODS: A retrospective review of 341 patients who had pulmonary edema fluid collected prospectively after the acute onset of pulmonary edema. All patients had serial samples of edema fluid and plasma collected over the first 8 h after intubation. RESULTS: Ten of the 341 patients with acute pulmonary edema were identified as having postobstructive pulmonary edema. The mean (+/- SD) edema fluid/plasma protein ratio in these patients was 0.54 +/- 0.15. The mean rate of alveolar fluid clearance over 8 h was 14.0 +/- 17.4% per hour. Nine of the 10 patients survived the hospitalization. CONCLUSION: Measurement of the edema fluid/plasma protein ratio and the presence of net alveolar fluid clearance in 10 patients with postobstructive pulmonary edema supports a hydrostatic mechanism for edema fluid formation. The predominantly fast rates of alveolar fluid clearance may explain the rapid resolution of clinical postobstructive pulmonary edema that is typically described. PMID- 17413052 TI - Asthma prevalence, family size, and birth order. AB - BACKGROUND: Asthma prevalence may be reduced in large families. The hygiene hypothesis suggests that older siblings protect their younger siblings from asthma through a modulating effect on the still-maturing immune system. If the hygiene hypothesis is correct, asthma prevalence should be inversely related to birth order. The objective of this study was to examine the relationship between asthma prevalence, and family size and birth order. METHODS: The medical records of 531,116 Israeli military conscripts were reviewed. The association between number of children in the family and the prevalence of asthma, and between birth order and the prevalence of asthma was assessed. Odds ratios for asthma by birth order and family size, adjusted for each other, were calculated. RESULTS: Asthma was diagnosed in 26,833 male subjects (8.6%) and 15,079 female subjects (6.9%). Asthma prevalence was inversely related to the number of children in the family (p < 0.001). Among subjects who were the only child in the family, the prevalence of asthma was 7.3%. The prevalence increased to 8.95% among subjects from families with three siblings, and then progressively decreased as the number of siblings increased, and reached a trough of 0.58% in conscripts from families of 15 to 20 siblings. Asthma prevalence was similar for all birth orders. CONCLUSIONS: In families with four or more children, asthma prevalence is inversely related to the number of children in the family. Asthma prevalence is similar for all birth orders. The similar asthma prevalence for all birth orders challenges the hygiene hypothesis as the mechanism for the decreased asthma prevalence in large families. PMID- 17413054 TI - Risk of progression to active tuberculosis among foreign-born persons with latent tuberculosis. AB - BACKGROUND: Increased risk for tuberculosis (TB) disease has been identified in foreign-born persons in the United States, particularly during the first 5 years after their arrival in the United States. This could be explained by undetected TB disease at entry, increased prevalence of latent TB infection (LTBI), increased progression from LTBI to TB, or a combination of these factors. METHODS: We performed a cluster analysis of TB cases in Boston and a case-control study of risk factors for TB with an unclustered isolate among Boston residents with LTBI to determine whether such persons have an increased risk for reactivation of disease. RESULTS: Of 321 case patients with TB seen between 1996 and 2000, 133 isolates were clustered and 188 were not. In multivariate analysis, foreign birth was associated with an unclustered isolate (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.2 to 3.8; p < 0.01), while being a close contact of a TB case was negatively associated (OR, 0.22; 95% CI, 0.07 to 0.73; p = 0.02). When 188 TB patients with unclustered isolates were compared to 188 age matched control subjects with LTBI, there was no association between the occurrence of TB and foreign birth (OR, 0.71; 95% CI, 0.42 to 1.3); among foreign born persons, there was no association between the occurrence of TB and being in the United States 5 SDs shorter than the mean QTc. CONCLUSIONS: The short QTc reported by an ECG computer was inaccurate and required manual correction. Short QT syndrome, defined as a QTc of < or = 300 ms, is rare. We were unable to find one patient among a population > 100,000 patients with a true QTc of < 300 ms. PMID- 17413056 TI - The effect of the 1995 heat wave in Chicago on all-cause and cause-specific mortality. AB - OBJECTIVES: We sought to reexamine the effects of the 1995 Chicago heat wave on all-cause and cause-specific mortality, including mortality displacement, using advanced time-series analysis methods. METHODS: We used Poisson regression with penalized regression splines to model excess mortality and mortality displacement over a 50-day period centered on the day in which the heat wave temperature peaked, adjusting for meteorological and other variables. We controlled for temporal trends by using daily mortality data during 1993-1997. We estimated relative risks (RRs) with reference to the first day of the 50-day period. RESULTS: We estimated that there were 692 excess deaths from June 21, 1995, to August 10, 1995; 26% of these deaths were owing to mortality displacement. RR for all-cause mortality on the day with peak mortality was 1.74 (95% confidence interval=1.67, 1.81). Risk of heat-related death was significantly higher among Blacks, and mortality displacement was substantially lower. CONCLUSIONS: The 1995 Chicago heat wave substantially effected all-cause and cause-specific mortality, but mortality displacement was limited. Mortality risks and displacement affected Blacks disproportionally. Appropriately targeted interventions may have a tangible effect on life expectancy. PMID- 17413057 TI - Public health investigation after the discovery of ricin in a South Carolina postal facility. AB - OBJECTIVES: In October 2003, a package containing ricin and a note threatening to poison water supplies was discovered in a South Carolina postal facility, becoming the first potential chemical terrorism event involving ricin in the United States. We examined the comprehensive public health investigation that followed and discuss the lessons learned from it. METHODS: An investigation consisting primarily of environmental sampling for ricin contamination, performance of health assessments on affected personnel, and local, regional, and national surveillance for ricin-associated illness. RESULTS: Laboratory analysis of 75 environmental sampling specimens revealed no ricin contamination. Health assessments of 36 affected employees were completed. Local surveillance initially identified 3 suspected cases, and national surveillance identified 399 outliers during the 2-week period after the incident. No confirmed cases of ricin associated illness were identified. CONCLUSIONS: A multifaceted and multidisciplinary approach is required for an effective public health response to a chemical threat such as ricin. The results of all of the described activities were used to determine that the facility was safe to reopen and that no public health threat existed. PMID- 17413058 TI - Benefits and barriers to electronic laboratory results reporting for notifiable diseases: the New York City Department of Health and Mental Hygiene experience. AB - OBJECTIVES: Despite national support for electronic laboratory reporting (ELR), the transition from paper to electronic reporting has been slow both nationally and locally. We assessed the ELR experience of New York City's surveillance programs to identify barriers to ELR implementation and generalizable lessons about automated electronic notifiable disease surveillance. METHODS: We conducted interviews with key staff of the New York City Department of Health and Mental Hygiene to evaluate ELR implementation. A review of paper and ELR disease reports enabled a comparison of the reporting systems. RESULTS: The completeness and timeliness of ELR were similar to, and sometimes better than, paper reporting for certain diseases. Incorporating electronic data into surveillance databases created new problems with data quality, shifted work demands, and required additional skills for data monitoring. ELR improved the handling of high-volume and time-sensitive diseases but did not completely automate reporting for diseases that required complicated assessments by staff. CONCLUSIONS: Although ELR streamlines data processing, electronic reporting has its own limitations. A more successful use of ELR can be achieved by understanding its strengths and limitations for different disease types. PMID- 17413059 TI - Encouraging compliance with quarantine: a proposal to provide job security and income replacement. AB - A human influenza virus is considered the most likely source of a pandemic in the near future. Quarantine has the potential to be the most effective measure for limiting the spread of infection. The major obstacles to compliance for those asked to enter quarantine include loss of income during quarantine and loss of employment after quarantine. We discuss current antidiscrimination and compensation laws, as well as options to expand coverage for quarantined individuals to encourage public cooperation by guaranteeing job security and providing income replacement. PMID- 17413060 TI - Adequacy of state capacity to address noncommunicable disease clusters in the era of environmental public health tracking. AB - OBJECTIVES: We sought to assess the capacity of state public health agencies to address noncommunicable disease clusters (NCCs) and to develop recommendations to enhance agencies' NCC response capacity. METHODS: We conducted an inventory of state public health agency Web sites and administered a Web-based survey of state health agency personnel to examine NCC capacity with respect to responsibility and authority, scope, protocols, trends in NCC investigations, and desired assistance. RESULTS: Twenty-six of the state agency Web sites listed an NCC contact, and 12 mentioned a cluster response team. Thirty-seven states completed the Web-based survey, all addressed cancer clusters, and 30 also responded to other NCCs, such as multiple sclerosis, amyotrophic lateral sclerosis, and autism. CONCLUSIONS: NCCs are of key concern to communities, and all of the survey respondents indicated that citizen reports were an impetus for investigations; yet, state-level capacity to address NCCs was inconsistent and disjointed. State agency personnel were committed to responding to NCC inquiries but were hampered by lack of personnel, resources, and prescribed protocols, as well as inadequate interagency communication. We offer recommendations to address these challenges. PMID- 17413061 TI - Disrupting the transmission of influenza a: face masks and ultraviolet light as control measures. AB - In the event of an influenza pandemic, where effective vaccine and antiviral drugs may be lacking, disrupting environmental transmission of the influenza virus will be the only viable strategy to protect the public. We discuss 2 such modalities, respirators (face masks) and ultraviolet (UV) light. Largely overlooked, the potential utility of each is underappreciated. The effectiveness of disposable face masks may be increased by sealing the edges of the mask to the face. Reusable masks should be stockpiled, because the supply of disposable masks will likely prove inadequate. UV light, directed overhead, may be beneficial in hospitals and nursing homes. PMID- 17413062 TI - Tsunami mortality estimates and vulnerability mapping in Aceh, Indonesia. AB - OBJECTIVES: We aimed to quantify tsunami mortality and compare approaches to mortality assessment in the emergency context in Aceh, Indonesia, where the impact of the 2004 tsunami was greatest. METHODS: Mortality was estimated using geographic information systems-based vulnerability models and demographic methods from surveys of tsunami-displaced populations. RESULTS: Tsunami mortality in Aceh as estimated by demographic models was 131066 and was similar to official figures of 128063; however, it was a conservative estimate of actual mortality and is substantially less than official estimates of 168561 presumed dead, which included those classified as missing. Tsunami impact was greatest in the district of Aceh Jaya, where an estimated 27.0% (n=23862) of the population perished; Aceh Besar and Banda Aceh were also severely affected, with mortality at 21.0% (n = 61 650) and 11.5% (n = 25 903), respectively. Mortality was estimated at 23.7% for the population at risk and 5.6% overall. CONCLUSIONS: Mortality estimates were derived using methodologies that can be applied in future disasters when predisaster demographic data are not available. Models could be useful in the early stages of disaster response by facilitating geographic targeting and management of humanitarian assistance. PMID- 17413063 TI - The role of the public health official in communicating public health information. AB - The prevailing views on the role of public health professionals refer to professionals in the academic world, without taking into account the fact that many public health professionals are government employees. For example, the American Public Health Association states that public health professionals play an active role in communicating public health information to nonscientific audiences, such as the general population or the mass media. We propose that public health officials have an important responsibility to promote the practice of public health. However, they must do so within the bureaucracy. Any actions that public health officials wish to take as advocates of particular public health activities should be carried out independent of their role as government officials. PMID- 17413064 TI - Evaluation of potassium iodide prophylaxis knowledge and nuclear emergency preparedness: New Jersey 2005. AB - The Nuclear Regulatory Commission requires states to consider including potassium iodide as a protective measure in the unlikely event of a major release of radioactivity from a nuclear power plant. We evaluated emergency preparedness knowledge, including proper potassium iodide use, among the general public and emergency responders located around New Jersey's nuclear power plants. We found that knowledge about responder chain of command, evacuation routes, and some aspects of potassium iodide usage was incomplete among the general public and emergency responders. PMID- 17413065 TI - Household water disinfection in hurricane-affected communities of Louisiana: implications for disaster preparedness for the general public. AB - OBJECTIVES: Thousands of Louisiana residents were asked to boil water because of widespread disruptions in electricity and natural gas services after Hurricane Rita. We sought to assess awareness of boil water orders and familiarity with household water disinfection techniques other than boiling. METHODS: We conducted a cross-sectional survey in randomly selected mobile home communities in Louisiana. RESULTS: We interviewed 196 respondents from 8 communities, which had boil water orders instituted. Of 97 who were home while communities were still under orders to boil water, 30 (31%) were aware of the orders and, of those, 24 (80%) said the orders were active while they were living at home; of the 24, 10 (42%) reported boiling water. Overall, 163 (83%) respondents were aware of a method of water disinfection at the household level: boiling (78%), chlorination (27%), and filtration (25%); 87% had a container of chlorine bleach at home. CONCLUSIONS: Few hurricane-affected respondents were aware of boil water orders and of alternate water disinfection techniques. Most had access to chlorine and could have practiced household chlorination if disruption in natural gas and electricity made boiling impossible. PMID- 17413066 TI - Ethics in a pandemic: a survey of the state pandemic influenza plans. AB - A pandemic of highly pathogenic influenza would threaten the lives of hundreds of thousands in the United States and confront governments and organizations, with ethical issues having wide-ranging implications. The Department of Health and Human Services and all states have published pandemic influenza plans. We analyzed the federal and state plans, available on the Internet, for evidence of ethical guidance as judged by the presence of ethical terms. The most striking finding was an absence of ethical language. Although some states acknowledged the need for ethical decisionmaking, very few prescribed how it should happen. If faced by a pandemic in the near future, we stand the risk of making many unjust and regrettable decisions. PMID- 17413067 TI - Psychological sequelae resulting from the 2004 Florida hurricanes: implications for postdisaster intervention. AB - OBJECTIVES: Data are limited regarding mental health effects of disasters such as hurricanes. We sought to determine the prevalence of and major risk factors associated with posttraumatic stress disorder (PTSD), generalized anxiety disorder, and major depressive episode 6 to 9 months after the 2004 Florida hurricanes. METHODS: Random-digit dialing was used to recruit a representative population sample of 1452 hurricane-affected adults. RESULTS: Posthurricane prevalence for PTSD was 3.6%, for generalized anxiety disorder was 5.5%, and for major depressive episode was 6.1%. Risk factors varied somewhat across disorders, with the exception of previous exposure to traumatic events, which increased risk of all negative outcomes. CONCLUSIONS: Storm exposure variables and displacement were associated primarily with PTSD. Notably, high social support in the 6 months preceding the hurricanes protected against all types of disorders. PMID- 17413068 TI - Trauma and stress response among Hurricane Katrina evacuees. AB - OBJECTIVES: Hurricane Katrina's impact on public health has been significant and multifaceted, with trauma-related psychological sequelae likely to result in a sizable burden of disease. Data were collected that assessed acute stress disorder (ASD) prevalence and factors related to ASD symptomatology among sheltered evacuees. METHODS: On days 12 to 19 after Katrina, evacuees at a major emergency shelter completed surveys that assessed demographics, Katrina-specific experiences, and ASD symptomatology. RESULTS: Sixty-two percent of the sample met ASD threshold criterion. Projections based on the predictive power of ASD to posttraumatic stress disorder (PTSD) suggest that 38% to 49% of the sample will meet PTSD criteria 2 years post-disaster. Female gender (odds ratio [OR] = 4.08), positive psychiatric history (OR=5.84), injury (OR=2.75), increased life-threat perception (OR=1.37), and decreased sense of personal control (OR=1.56) were significantly related to ASD. Black race was associated with greater symptom severity (B=7.85, SE[B]=3.50). CONCLUSIONS: Katrina-related trauma and its psychological sequelae will remain a significant public health issue for years to come. The identification of several vulnerability factors related to ASD and PTSD provides a brief sketch of those at greatest risk. PMID- 17413069 TI - Disaster planning and risk communication with vulnerable communities: lessons from Hurricane Katrina. AB - OBJECTIVES: We studied the experience of Hurricane Katrina evacuees to better understand factors influencing evacuation decisions in impoverished, mainly minority communities that were most severely affected by the disaster. METHODS: We performed qualitative interviews with 58 randomly selected evacuees living in Houston's major evacuation centers from September 9 to 12, 2005. Transcripts were content analyzed using grounded theory methodology. RESULTS: Participants were mainly African American, had low incomes, and were from New Orleans. Participants' strong ties to extended family, friends, and community groups influenced other factors affecting evacuation, including transportation, access to shelter, and perception of evacuation messages. These social connections cut both ways, which facilitated and hindered evacuation decisions. CONCLUSIONS: Effective disaster plans must account for the specific obstacles encountered by vulnerable and minority communities. Removing the more apparent obstacles of shelter and transportation will likely be insufficient for improving disaster plans for impoverished, minority communities. The important influence of extended families and social networks demand better community-based communication and preparation strategies. PMID- 17413070 TI - Legal tools for preparedness and response: variation in quarantine powers among the 10 most populous US states in 2004. AB - From April 2004 through December 2004, we reviewed the express legal authorities of the 10 most populous US states to restrict the movement of persons to control communicable diseases. All 10 of the states possessed express legal authority to quarantine and isolate individuals, but the laws varied substantially. In the absence of declared emergencies, only 4 states had express authority to conduct area quarantine, and only 2 states had express authority to conduct group quarantine. During declared emergencies, 7 states had additional authorities for area quarantine. Express authorities are only part of states' legal powers to employ such movement restrictions, but substantial variation in express authorities across states could present potential challenges for the coordination of large national or regional epidemics. PMID- 17413071 TI - Optimizing severe acute respiratory syndrome response strategies: lessons learned from quarantine. AB - Taiwan used quarantine as 1 of numerous interventions implemented to control the outbreak of severe acute respiratory syndrome in 2003. From March 18 to July 31, 2003, 147,526 persons were placed under quarantine. Quarantining only persons with known exposure to people infected with severe acute respiratory syndrome could have reduced the number of persons quarantined by approximately 64%. Focusing quarantine efforts on persons with known or suspected exposure can greatly decrease the number of persons placed under quarantine, without substantially compromising its yield and effectiveness. PMID- 17413072 TI - A review of model public health laws. AB - Model public health laws (public health laws or private policies publicly recommended by at least 1 organization for adoption by government bodies or by specified private entities) are promoted as exemplary. We assessed the information sponsors of model public health laws provide on the methods used in developing their models and on their models' adoption and effectiveness. Through a systematic search, we identified 107 model public health laws published from 1907 to 2004. As of our assessment in 2005, only 18 (44%) of the sponsors presented any information on the procedures and evidence used in developing their model public health laws; information on adoption was provided for only 7 (6.5%) model laws. No sponsors provided information on model effectiveness. We recommend sponsors improve their disclosure of information about the methods and evidence used in developing model public health laws and about their adoption and effectiveness. PMID- 17413073 TI - Vision for a global registry of anticipated public health studies. AB - In public health, the generation, management, and transfer of knowledge all need major improvement. Problems in generating knowledge include an imbalance in research funding, publication bias, unnecessary studies, adherence to fashion, and undue interest in novel and immediate issues. Impaired generation of knowledge, combined with a dated and inadequate process for managing knowledge and an inefficient system for transferring knowledge, mean a distorted body of evidence available for decisionmaking in public health. This article hopes to stimulate discussion by proposing a Global Registry of Anticipated Public Health Studies. This prospective, comprehensive system for tracking research in public health could help enhance collaboration and improve efficiency. Practical problems must be discussed before such a vision can be further developed. PMID- 17413074 TI - Optimal shift duration and sequence: recommended approach for short-term emergency response activations for public health and emergency management. AB - Since September 11, 2001, and the consequent restructuring of the US preparedness and response activities, public health workers are increasingly called on to activate a temporary round-the-clock staffing schedule. These workers may have to make key decisions that could significantly impact the health and safety of the public. The unique physiological demands of rotational shift work and night shift work have the potential to negatively impact decisionmaking ability. A responsible, evidence-based approach to scheduling applies the principles of circadian physiology, as well as unique individual physiologies and preferences. Optimal scheduling would use a clockwise (morning-afternoon-night) rotational schedule: limiting night shifts to blocks of 3, limiting shift duration to 8 hours, and allowing 3 days of recuperation after night shifts. PMID- 17413075 TI - Expanding Bioshield: a call for caution. AB - The Bioshield Initiative seeks to stimulate development of new drugs and vaccines to prevent and treat diseases caused by biological agents likely to be used by terrorists, and recent proposals have sought to expand patent protections in this context. To the extent that patent protections are needed as part of the incentive structure for new drug and vaccine development, it may be better to strengthen patent protections on nonterror-related drugs and vaccines to avoid production capacity problems, as well as follow-up research and development problems. However, at the same time, both practical and ethical considerations argue that the good that might be achieved through expanded patent protections come at costs that make this strategy unacceptable. PMID- 17413076 TI - Evidence and effectiveness in decisionmaking for quarantine. AB - When public health decisionmakers turned to quarantine during the recent severe acute respiratory syndrome (SARS) epidemic, difficult questions were raised about the legitimacy and acceptability of restrictive measures to attain public health goals. SARS also brought to light how scientific uncertainty can permeate public health decisionmaking, leading us to think about the relationship between the adequacy of evidence of the effectiveness of an intervention and its role in the justification of public health action. In this article, we critically examine the role of evidence and effectiveness in decision-making for quarantine. It is our contention that the effectiveness of a public health intervention should not be defined exclusively in (absolute and objective) scientific terms but rather conceptualized relationally and normatively in public health decisionmaking. PMID- 17413077 TI - Preparedness for infectious threats: public-private partnership to develop an affordable vaccine for an emergent threat: the trivalent Neisseria meningitidis ACW135 polysaccharide vaccine. AB - With the emergence of epidemic Neisseria meningitidis W135 meningitis in Burkina Faso during early 2002, the public health community was faced with the challenge of providing access to an appropriate and affordable vaccine in time for the upcoming 2003 epidemic season. Recognizing the implications of the emergent threat, the World Health Organization developed a strategy, established a public private partnership to provide the needed vaccine, and then ensured that a stockpile was available for future use. The trivalent N meningitidis ACW135 polysaccharide vaccine that resulted is now one of the primary tools for epidemic response in African meningitis belt countries. It will remain so for the foreseeable future and until appropriate and affordable conjugate vaccines become part of national immunization programs in the region. PMID- 17413079 TI - Ready or not? PMID- 17413078 TI - Conceptualizing and defining public health emergency preparedness. PMID- 17413080 TI - Prepared to serve public health. PMID- 17413081 TI - Sports utility vehicles and vulnerable road users. PMID- 17413082 TI - Use of electronic health records in disaster response: the experience of Department of Veterans Affairs after Hurricane Katrina. AB - OBJECTIVES: We describe electronic health data use by the Department of Veterans Affairs (VA) in the month after Katrina, including supporting technologies, the extent and nature of information accessed, and lessons learned. METHODS: We conducted a retrospective study using cross-sectional panels of data collected sequentially over time. RESULTS: By September 30, 2005, clinical data were accessed electronically for at least 38% (14941 of 39910) of patients cared for prior to Hurricane Katrina by New Orleans-area VA medical facilities. Approximately 1000 patients per day had data accessed during the month following Hurricane Katrina, a rate approximately two thirds of pre-Katrina values. Health care data were transmitted to more than 200 sites in 48 states and to at least 2300 users. CONCLUSIONS: The VA electronic health records supported continuity of care for evacuated veterans after Katrina. Our findings suggest that pharmacy and laboratory computerization alone will not be sufficient for future disaster support systems. PMID- 17413083 TI - The law and emergencies: surveillance for public health-related legal issues during Hurricanes Katrina and Rita. AB - Law influenced every aspect of the public health response to Hurricanes Katrina and Rita, from evacuation orders, to waivers of medical licensing requirements, to the clean-up of public health threats on private property. We used public health surveillance of news reports to identify and characterize legal issues arising during the disaster response in 5 Gulf Coast states. Data collected from news reports of the events in real time were followed-up by interviews with selected state legal and emergency management officials. Our analysis indicates the value of surveillance during and after emergency responses in identifying public health-related legal issues and helps to inform the strengthening of legal preparedness frameworks for future disasters. PMID- 17413084 TI - The courts, public health, and legal preparedness. AB - The judicial branch's key roles, as guardian of civil liberties and protector of the rule of law, can be acutely relevant during public health emergencies when courts may need to issue orders authorizing actions to protect public health or restraining public health actions that are determined to unduly interfere with civil rights. Legal preparedness for public health emergencies, therefore, necessitates an understanding of the court system and how courts are involved in public health issues. In this article we briefly describe the court system and then focus on what public health practitioners need to know about the judicial system in a public health emergency, including the courts' roles and the consequent need to keep courts open during emergencies. PMID- 17413085 TI - Mutual aid agreements: essential legal tools for public health preparedness and response. AB - Mutual aid is the sharing of supplies, equipment, personnel, and information across political boundaries. States must have agreements in place to ensure mutual aid to facilitate effective responses to public health emergencies and to detect and control potential infectious disease outbreaks. The 2005 hurricanes triggered activation of the Emergency Management Assistance Compact (EMAC), a mutual aid agreement among the 50 states, the District of Columbia, Puerto Rico, and the US Virgin Islands. Although EMAC facilitated the movement of an unprecedented amount of mutual aid to disaster areas, inadequacies in the response demonstrated a need for improvement. Mutual aid may also be beneficial in circumstances where EMAC is not activated. We discuss the importance of mutual aid, examine obstacles, and identify legal "gaps" that must be filled to strengthen preparedness. PMID- 17413086 TI - African Americans' decisions not to evacuate New Orleans before Hurricane Katrina: a qualitative study. AB - OBJECTIVES: We examined the psychosocial and personal factors that influenced African Americans' decision not to evacuate New Orleans, La, before Hurricane Katrina's landfall. METHODS: We conducted 6 focus groups with 53 African Americans from New Orleans who were evacuated to Columbia, SC, within 2 months of Hurricane Katrina. RESULTS: The major themes identified related to participants' decision to not evacuate were as follows: (1) perceived susceptability, including optimism about the outcome because of riding out past hurricanes at home and religious faith; (2) perceived severity of the hurricane because of inconsistent evacuation orders; (3) barriers because of financial constraints and neighborhood crime; and (4) perceived racism and inequities. CONCLUSIONS: Federal, state, and local government disaster preparedness plans should specify criteria for timely evacuation orders, needed resources, and their allocation (including a decentralized distribution system for cash or vouchers for gas and incidentals during evacuation) and culturally sensitive logistic planning for the evacuation of minority, low-income, and underserved communities. Perceptions of racism and inequities warrant further investigation. PMID- 17413087 TI - Soybeans or soymilk: does it make a difference for cardiovascular protection? Does it even matter? PMID- 17413088 TI - The importance of fish and docosahexaenoic acid in Alzheimer disease. PMID- 17413089 TI - An overview of evidence for a causal relation between iron deficiency during development and deficits in cognitive or behavioral function. AB - This review, intended for a broad scientific readership, summarizes evidence relevant to whether a causal relation exists between dietary iron deficiency with (ID+A) or without (ID-A) anemia during development and deficits in subsequent cognitive or behavioral performance. An overview of expert opinion and major evidence in humans and animals is provided. Cognitive and behavioral effects observed in humans with ID-A and in animals with ID+/-A are provided in tables. The degree to which 5 conditions of causality are satisfied and whether deleterious effects of ID-A might be expected to occur are discussed. On the basis of the existing literature, our major conclusions are as follows. Although most of the 5 conditions of causality (association, plausible biological mechanisms, dose response, ability to manipulate the effect, and specificity of cause and effect) are partially satisfied in humans, animals, or both, a causal connection has not been clearly established. In animals, deficits in motor activity are consistently associated with severe ID+A, but adverse effects on performance in tests that target cognitive function have not been clearly shown. Resistance to iron treatment was observed in most trials of children <2 y of age with ID+A, but not in older children. Similar observations were made in rodents when ID+A occurred before rather than after weaning. In children >2 y of age and in adolescents with ID-A, evidence suggests cognitive or behavioral deficits; however, the surprisingly small number of studies conducted in either humans or animals prevents a thorough assessment. PMID- 17413090 TI - Consideration of betaine and one-carbon sources of N5-methyltetrahydrofolate for use in homocystinuria and neural tube defects. AB - A major focus in attempts to ameliorate homocystinuria and neural tube defects is supplementation of the diet with B vitamins. The metabolic defect in these cases may be due in part to a deficiency of methyl groups. B vitamin supplementation supports the need for enzyme cofactors but cannot provide substrate in the form of methyl groups. l-Methionine is an essential amino acid and is required for protein synthesis, but it also plays a unique role in metabolism as S adenosylmethionine, which is the primary methyl donor in metabolism. The observation that l-homocysteine, which is produced in the metabolism of l methionine, is remethylated 2-4 times before it is destroyed is key to understanding the possibility of a methyl group deficiency. This suggests that the requirement for methyl groups (ie, S-adenosylmethionine) may be 2-4 times that for methionine in support of protein synthesis. l-Homocysteine can be remethylated to form l-methionine by betaine or N(5)-methyltetrahydrofolate. Betaine and one-carbon sources that lead to the production of N(5) methyltetrahydrofolate and the remethylation of l-homocysteine to form l methionine should be considered along with B vitamin supplementation in the treatment of homocystinuria and neural tube defects. PMID- 17413091 TI - Effect of the melanocortin-3 receptor C17A and G241A variants on weight loss in childhood obesity. AB - BACKGROUND: The central melanocortin system is critical for the long-term regulation of energy homeostasis. Melanocortin-3 receptor (MC3R) knock-out mice, despite being hypophagic, have increased fat mass and higher feed efficiency than do their wild-type littermates. OBJECTIVE: The aim was to evaluate whether, in childhood obesity, MC3R variants are associated with changes in fatness reduction as a consequence of a weight-reduction program. DESIGN: Molecular screening of the MC3R coding region in 184 obese children, 77 girls and 107 boys [x (+/-SEM) body mass index (BMI; in kg/m(2)) z score: 3.3 +/- 2.3; age 9.2 +/- 2 y], was performed. BMI was evaluated at baseline and after 6 and 12 mo of the weight loss program. RESULTS: No new mutations were found. Two previously described polymorphisms, C17A (Thr6Lys) and G241A (Val81Ile), were observed in 20 patients in almost complete linkage disequilibrium. No significant differences in BMI z scores were observed at baseline of the weight-loss program between the genotypes; however, at follow-up, heterozygotes showed a significantly higher BMI z score (P = 0.03). When the patients were divided according to the amount of weight lost, a higher prevalence of heterozygotes was observed among subjects who lowered their BMI z score <1.5 (P = 0.03). CONCLUSION: These results suggest a gene-diet interaction between the MC3R C17A and G241A variants and a weight loss program for the ability to lose weight in childhood obesity. PMID- 17413092 TI - Long-term weight losses associated with prescription of higher physical activity goals. Are higher levels of physical activity protective against weight regain? AB - BACKGROUND: High levels of exercise may be necessary for long-term maintenance of weight loss. OBJECTIVE: We aimed to determine in a randomized prospective design whether encouraging 2500 kcal physical activity/wk produced greater 30-mo weight losses than did the standard 1000 kcal physical activity/wk prescription. DESIGN: Overweight adults (n = 202) were randomly assigned to either 18 mo of standard behavioral treatment (SBT) with an exercise goal of 1000 kcal/wk or a high physical activity (HPA) treatment with a goal of 2500 kcal/wk. The HPA treatment included all procedures in the SBT plus encouragement to recruit 1-3 exercise partners and small-group counseling with an exercise coach. Participants were followed for 30 mo. RESULTS: The HPA group achieved significantly greater exercise levels and weight losses than did the SBT group at 12 and 18 mo (P < 0.01). Weight losses did not differ significantly at 30 mo: 0.90 +/- 8.9 and 2.86 +/- 8.6 kg for the SBT and HPA groups, respectively (P = 0.16). At 30 mo, average exercise levels no longer differed significantly between groups (1390 and 1696 kcal/wk, respectively; P > 0.10). Participants sustaining high exercise levels (>2500 kcal/wk) for 30 mo had significantly (P < 0.001) greater 30-mo weight loss than did those exercising less (12 +/- 8.8 and 0.8 +/- 8.1 kg, respectively). CONCLUSIONS: Although participants in the HPA group sustained the 2500-kcal activity goal during the 18-mo treatment, activity declined once treatment ended, which resulted in no between-group differences in activity or weight loss at 2.5 y. Participants who reported continuing to engage in high levels of exercise maintained a significantly larger weight loss. PMID- 17413093 TI - Effect of soy protein from differently processed products on cardiovascular disease risk factors and vascular endothelial function in hypercholesterolemic subjects. AB - BACKGROUND: The magnitude of the effect of soy protein on lipoprotein concentrations is variable. This discordance is likely attributable to the various forms of soy protein used and to unrecognized shifts in dietary fatty acid, cholesterol, and fiber. OBJECTIVE: The objective was to evaluate the effect of soybean processing as well as soy consumption relative to animal protein, independent of alterations in major dietary variables, on cardiovascular disease risk factors and vascular endothelial function. DESIGN: Twenty-eight hypercholesterolemic subjects (LDL cholesterol >/=3.36 mmol/L) aged >50 y consumed each of 4 diets for 6-wk periods according to a randomized crossover design. The diets [55% of energy as carbohydrate, 30% of energy as fat, and 15% of energy as protein-7.5% of energy as experimental protein (37.5 g/d)] were designed to contain products made from either whole soybeans, soyflour, or soymilk and were compared with a diet containing an equivalent amount of animal protein (meat, chicken, and dairy products). The cholesterol, fiber, and fatty acid profiles of the diets were equalized. All food and drink were provided, and body weight was maintained throughout the study. RESULTS: No significant differences in blood pressure, vascular endothelial function, or total cholesterol, VLDL-cholesterol, triacylglycerol, apolipoprotein B, or C-reactive protein concentrations were observed between the diets. Consumption of the soymilk diet resulted in a modest decrease (4%) in LDL-cholesterol concentrations compared with the animal-protein and soyflour diets (P < 0.05) and higher HDL cholesterol (1%) and apolipoprotein A-I (2%) concentrations compared with the soybean and soyflour diets (P < 0.05). CONCLUSIONS: The results suggest that the consumption of differently processed soy-based products and different types of protein (animal and soy) has little clinical effect on cardiovascular disease risk factors, including peripheral endothelial function, when other major dietary variables are held constant. PMID- 17413094 TI - Constitutional thinness and lean anorexia nervosa display opposite concentrations of peptide YY, glucagon-like peptide 1, ghrelin, and leptin. AB - BACKGROUND: Food intake is controlled by the arcuate nucleus through integration of peripheral hormonal signals such as leptin, ghrelin, peptide YY (PYY), and glucagon-like peptide 1 (GLP-1). The most common condition resulting in underweight young women in the developed world is restrictive anorexia nervosa (AN). However, constitutional thinness (CT) is also known to exist in the same low-weight range. Women with CT have normal menstrual periods and do not have the psychological or hormonal features of AN. Little is currently known about regulation of food intake in subjects with CT. OBJECTIVE: We tested the hypothesis that concentrations of leptin, ghrelin, PYY, and GLP-1 in persons with AN are significantly different from those in persons with CT. DESIGN: Concentrations of PYY, GLP-1, ghrelin, and leptin were measured in 3 groups of young women: normal weight (n = 7), CT (n = 10), and AN (n = 12). Samples were collected every 4 h for 24 h. RESULTS: PYY concentrations were significantly higher in CT subjects than in AN or control subjects. GLP-1 concentrations were significantly higher in AN than in CT subjects, whereas ghrelin was significantly higher in AN subjects than in control and CT subjects. CT subjects had the lowest ghrelin concentrations. Leptin concentrations were significantly lower in AN subjects. PYY and leptin circadian variations were not significantly different between CT and control subjects, whereas these profiles were blunted in AN subjects. CONCLUSIONS: Orexigenic and anorexigenic hormones in CT contrast with an adaptative profile characterizing AN. The hormones appear to be valuable biomarkers for distinguishing these 2 categories of severely underweight subjects. PMID- 17413095 TI - Premenopausal overweight women do not lose bone during moderate weight loss with adequate or higher calcium intake. AB - BACKGROUND: Weight loss is associated with bone loss, but this has not been examined in overweight premenopausal women. OBJECTIVE: The aim of this study was to assess whether overweight premenopausal women lose bone with moderate weight loss at recommended or higher than recommended calcium intakes. DESIGN: Overweight premenopausal women [n = 44; x (+/-SD) age: 38 +/- 6.4 y; body mass index (BMI): 27.7 +/- 2.1 kg/m(2)] were randomly assigned to either a normal (1 g/d) or high (1.8 g/d) calcium intake during 6 mo of energy restriction [weight loss (WL) groups] or were recruited for weight maintenance at 1 g Ca/d intake. Regional bone mineral density and content were measured by dual-energy X-ray absorptiometry, and markers of bone turnover were measured before and after weight loss. True fractional calcium absorption (TFCA) was measured at baseline and during caloric restriction by using a dual-stable calcium isotope method. RESULTS: The WL groups lost 7.2 +/- 3.3% of initial body weight. No significant decrease in BMD or rise in bone turnover was observed with weight loss at normal or high calcium intake. The group that consumed high calcium showed a strong relation (r = 0.71) between increased femoral neck bone mineral density and increased serum 25-hydroxyvitamin D. No significant effect of weight loss on TFCA was observed, and the total calcium absorbed was adequate at 238 +/- 81 and 310 +/- 91 mg/d for the normal- and high-calcium WL groups, respectively. CONCLUSION: Overweight premenopausal women do not lose bone during weight loss at the recommended calcium intake, which may be explained by sufficient amounts of absorbed calcium. PMID- 17413096 TI - A controlled trial of reduced meal frequency without caloric restriction in healthy, normal-weight, middle-aged adults. AB - BACKGROUND: Although consumption of 3 meals/d is the most common pattern of eating in industrialized countries, a scientific rationale for this meal frequency with respect to optimal health is lacking. A diet with less meal frequency can improve the health and extend the lifespan of laboratory animals, but its effect on humans has never been tested. OBJECTIVE: A pilot study was conducted to establish the effects of a reduced-meal-frequency diet on health indicators in healthy, normal-weight adults. DESIGN: The study was a randomized crossover design with two 8-wk treatment periods. During the treatment periods, subjects consumed all of the calories needed for weight maintenance in either 3 meals/d or 1 meal/d. RESULTS: Subjects who completed the study maintained their body weight within 2 kg of their initial weight throughout the 6-mo period. There were no significant effects of meal frequency on heart rate, body temperature, or most of the blood variables measured. However, when consuming 1 meal/d, subjects had a significant increase in hunger; a significant modification of body composition, including reductions in fat mass; significant increases in blood pressure and in total, LDL-, and HDL-cholesterol concentrations; and a significant decrease in concentrations of cortisol. CONCLUSIONS: Normal-weight subjects are able to comply with a 1 meal/d diet. When meal frequency is decreased without a reduction in overall calorie intake, modest changes occur in body composition, some cardiovascular disease risk factors, and hematologic variables. Diurnal variations may affect outcomes. PMID- 17413097 TI - Is higher dairy consumption associated with lower body weight and fewer metabolic disturbances? The Hoorn Study. AB - BACKGROUND: Dairy consumption has been postulated to reduce the risk of obesity and metabolic disturbances. OBJECTIVE: The aim of this study was to evaluate the associations of dairy consumption with body weight and other components of the metabolic syndrome. DESIGN: We used cross-sectional data for 2064 men and women aged 50-75 y who participated in the Hoorn Study. The metabolic syndrome was defined according to the National Cholesterol Education Program Expert Panel. Dairy consumption was assessed by using a semiquantitative food-frequency questionnaire. RESULTS: The median consumption of total dairy products was 4.1 servings/d. After adjustment for potential confounders (ie, dietary factors, physical activity, smoking, income, educational level, and antihypertensive medication), total dairy consumption was significantly associated with lower diastolic blood pressure (beta +/- SE: -0.31 +/- 0.12 mm Hg/serving) and higher fasting glucose concentrations (0.04 +/- 0.02 mmol/L per serving), but not with body weight or other metabolic variables (ie, lipids, postload glucose, or insulin). When different dairy products were distinguished, borderline significant (P < 0.10) inverse associations were observed for dairy desserts, milk, and yogurt with systolic (-1.26 +/- 0.58, -0.57 +/- 0.34, and -1.28 +/- 0.74 mm Hg/serving, respectively) and diastolic (-0.58 +/- 0.31, -0.57 +/- 0.18, and -0.35 +/- 0.40 mm Hg/serving, respectively) blood pressure, whereas cheese consumption was positively associated with body mass index (0.15 +/- 0.08/serving). CONCLUSION: In an elderly Dutch population, higher dairy consumption was not associated with lower weight or more favorable levels of components of the metabolic syndrome, except for a modest association with lower blood pressure. PMID- 17413098 TI - Metabolic effects of amino acid mixtures and whey protein in healthy subjects: studies using glucose-equivalent drinks. AB - BACKGROUND: Milk protein, in particular the whey fraction, has been shown to display insulinotrophic properties in healthy persons and persons with type 2 diabetes. In parallel to the hyperinsulinemia, a pronounced postprandial rise of certain amino acids and of glucose-dependent insulinotrophic polypeptide (GIP) was observed in plasma. OBJECTIVE: The objective of the study was to determine to what extent the insulinotrophic properties of whey could be simulated by specific amino acid mixtures. DESIGN: Twelve healthy volunteers were served drinks consisting of pure glucose (reference drink) or glucose supplemented with free amino acids or whey proteins (test drinks). RESULTS: A test drink with the branched-chain amino acids isoleucine, leucine, and valine resulted in significantly higher insulin responses than did the glucose reference. A drink containing glucose and leucine, isoleucine, valine, lysine, and threonine mimicked the glycemic and insulinemic responses seen after whey ingestion. With consumption of this drink, the glucose area under the curve (AUC) was 44% smaller (P < 0.05) and the insulin AUC was 31% larger (NS) than with consumption of the reference drink. With consumption of the whey drink, the AUCs were 56% smaller (glucose; P < 0.05) and 60% larger (insulin; P < 0.05), respectively, than with the reference drink. The whey drink was accompanied by an 80% greater GIP response (P < 0.05), whereas the drinks containing free amino acids did not significantly affect GIP secretion. CONCLUSION: A mixture of leucine, isoleucine, valine, lysine, and threonine resulted in glycemic and insulinemic responses closely mimicking those seen after whey ingestion in the absence of an additional effect of GIP and glucagon-like peptide 1. PMID- 17413099 TI - Resistance training and dietary protein: effects on glucose tolerance and contents of skeletal muscle insulin signaling proteins in older persons. AB - BACKGROUND: Resistance training (RT) and dietary protein independently influence indexes of whole-body glucose control, though their synergistic effects have not yet been documented. OBJECTIVE: This study assessed the influence of dietary protein intake on RT-induced changes in systemic glucose tolerance and the contents of skeletal muscle insulin signaling proteins in healthy older persons. DESIGN: Thirty-six older men and women (age: 61 +/- 1 y) performed RT (3 times/wk for 12 wk) and consumed either 0.9 g protein . kg(-1) . d(-1) [lower-protein (LP) group; approximately 112% of the Recommended Dietary Allowance (RDA)] or 1.2 g protein . kg(-1) . d(-1) [higher-protein (HP) group; approximately 150% of the RDA]; the HP group consumed more total, egg, and dairy proteins. RESULTS: After RT, body weight was unchanged; whole-body protein and water masses increased, and fat mass decreased with no significantly different responses observed between the LP and HP groups. The RT-induced improvement in oral glucose tolerance (decreased area under the curve, AUC) was not significantly different between the groups (LP: -28%; HP: -25%). The insulin (-21%) and C-peptide (-14%) AUCs decreased in the LP group but did not change significantly in the HP group. Skeletal muscle insulin receptor, insulin receptor substrate-1, and Akt contents were unchanged, and the amount of atypical protein kinase C zeta/lambda (aPKC zeta/lambda), a protein involved with insulin signaling, increased 56% with RT, independent of protein intake. CONCLUSION: These results support the hypothesis that older persons who consume adequate or moderately high amounts of dietary protein can use RT to improve body composition, oral glucose tolerance, and skeletal muscle aPKC zeta/lambda content without a change in body weight. PMID- 17413100 TI - Effect on 24-h energy expenditure of a moderate-fat diet high in monounsaturated fatty acids compared with that of a low-fat, carbohydrate-rich diet: a 6-mo controlled dietary intervention trial. AB - BACKGROUND: Dietary fat has a lower thermogenic effect than does carbohydrate. A moderate-fat diet, high in monounsaturated fatty acids (MUFA diet), may decrease energy expenditure (EE) and thereby induce weight gain. OBJECTIVE: We aimed to compare changes in 24-h EE and substrate oxidation after a 6-mo controlled dietary intervention with either a MUFA or a low-fat (LF) diet. DESIGN: Twenty seven overweight [body mass index (in kg/m(2)): 28.1 +/- 0.4] nondiabetic subjects aged 18-36 y followed an 8-wk low-calorie diet and a 2-wk weight stabilizing diet and then were randomly assigned to a MUFA (n = 12) or LF (n = 15) diet for 6 mo. Substrate oxidation and 24-h EE were measured by whole-body indirect calorimetry. The first measurement (0 mo) was taken during the weight stabilizing diet, and the second measurement was taken after the 6-mo intervention. RESULTS: A tendency was seen toward a lower 24-h EE with the MUFA than with the LF diet (P = 0.0675), but this trend did not remain after adjustment for the initial loses of fat mass and fat-free mass (P = 0.2963). Meal induced thermogenesis was significantly (P < 0.05) lower with the MUFA than with the LF diet, but no time x treatment interaction was found. A significant (P = 0.0456) treatment x time interaction was found for spontaneous physical activity. CONCLUSION: Despite a slightly lower meal-induced thermogenesis, the MUFA diet had an effect on 24-h EE that was not significantly different from that of the LF diet after a 6-mo controlled dietary intervention. PMID- 17413101 TI - Long-term effects of 2 energy-restricted diets differing in glycemic load on dietary adherence, body composition, and metabolism in CALERIE: a 1-y randomized controlled trial. AB - BACKGROUND: There remains no consensus about the optimal dietary composition for sustained weight loss. OBJECTIVE: The objective was to examine the effects of 2 dietary macronutrient patterns with different glycemic loads on adherence to a prescribed regimen of calorie restriction (CR), weight and fat loss, and related variables. DESIGN: A randomized controlled trial (RCT) of diets with a high glycemic load (HG) or a low glycemic load (LG) at 30% CR was conducted in 34 healthy overweight adults with a mean (+/-SD) age of 35 +/- 6 y and body mass index (kg/m(2)) of 27.6 +/- 1.4. All food was provided for 6 mo in diets controlled for confounding variables, and subjects self-administered the plans for 6 additional months. Primary and secondary outcomes included energy intake measured by doubly labeled water, body weight and fatness, hunger, satiety, and resting metabolic rate. RESULTS: All groups consumed significantly less energy during CR than at baseline (P < 0.01), but changes in energy intake, body weight, body fat, and resting metabolic rate did not differ significantly between groups. Both groups ate more energy than provided (eg, 21% and 28% CR at 3 mo and 16% and 17% CR at 6 mo with HG and LG, respectively). Percentage weight change at 12 mo was -8.04 +/- 4.1% in the HG group and -7.81 +/- 5.0% in the LG group. There was no effect of dietary composition on changes in hunger, satiety, or satisfaction with the amount and type of provided food during CR. CONCLUSIONS: These findings provide more detailed evidence to suggest that diets differing substantially in glycemic load induce comparable long-term weight loss. PMID- 17413102 TI - Consumption of fluid skim milk promotes greater muscle protein accretion after resistance exercise than does consumption of an isonitrogenous and isoenergetic soy-protein beverage. AB - BACKGROUND: Resistance exercise leads to net muscle protein accretion through a synergistic interaction of exercise and feeding. Proteins from different sources may differ in their ability to support muscle protein accretion because of different patterns of postprandial hyperaminoacidemia. OBJECTIVE: We examined the effect of consuming isonitrogenous, isoenergetic, and macronutrient-matched soy or milk beverages (18 g protein, 750 kJ) on protein kinetics and net muscle protein balance after resistance exercise in healthy young men. Our hypothesis was that soy ingestion would result in larger but transient hyperaminoacidemia compared with milk and that milk would promote a greater net balance because of lower but prolonged hyperaminoacidemia. DESIGN: Arterial-venous amino acid balance and muscle fractional synthesis rates were measured in young men who consumed fluid milk or a soy-protein beverage in a crossover design after a bout of resistance exercise. RESULTS: Ingestion of both soy and milk resulted in a positive net protein balance. Analysis of area under the net balance curves indicated an overall greater net balance after milk ingestion (P < 0.05). The fractional synthesis rate in muscle was also greater after milk consumption (0.10 +/- 0.01%/h) than after soy consumption (0.07 +/- 0.01%/h; P = 0.05). CONCLUSIONS: Milk-based proteins promote muscle protein accretion to a greater extent than do soy-based proteins when consumed after resistance exercise. The consumption of either milk or soy protein with resistance training promotes muscle mass maintenance and gains, but chronic consumption of milk proteins after resistance exercise likely supports a more rapid lean mass accrual. PMID- 17413103 TI - Carotene-rich plant foods ingested with minimal dietary fat enhance the total body vitamin A pool size in Filipino schoolchildren as assessed by stable-isotope dilution methodology. AB - BACKGROUND: Strategies for improving the vitamin A status of vulnerable populations are needed. OBJECTIVE: We studied the influence of the amounts of dietary fat on the effectiveness of carotene-rich plant foods in improving vitamin A status. DESIGN: Schoolchildren aged 9-12 y were fed standardized meals 3 times/d, 5 d/wk, for 9 wk. The meals provided 4.2 mg provitamin A carotenoids/d (mainly beta-carotene) from yellow and green leafy vegetables [carrots, pechay (bok choy), squash, and kangkong (swamp cabbage)] and 7, 15, or 29 g fat/d (2.4, 5, or 10 g fat/meal) in groups A, B, and C (n = 39, 39, and 38, respectively). Other self-selected foods eaten were recorded daily. Before and after the intervention, total-body vitamin A pool sizes and liver vitamin A concentrations were measured with the deuterated-retinol-dilution method; serum retinol and carotenoid concentrations were measured by HPLC. RESULTS: Similar increases in mean serum beta-carotene (5-fold), alpha-carotene (19-fold), and beta cryptoxanthin (2-fold) concentrations; total-body vitamin A pool size (2-fold); and liver vitamin A (2-fold) concentrations were observed after 9 wk in the 3 study groups; mean serum retinol concentrations did not change significantly. The total daily beta-carotene intake from study meals plus self-selected foods was similar between the 3 groups and was 14 times the usual intake; total fat intake was 0.9, 1.4, or 2.0 times the usual intake in groups A, B, and C, respectively. The overall prevalence of low liver vitamin A (<0.07 mumol/g) decreased from 35% to 7%. CONCLUSIONS: Carotene-rich yellow and green leafy vegetables, when ingested with minimal fat, enhance serum carotenoids and the total-body vitamin A pool size and can restore low liver vitamin A concentrations to normal concentrations. PMID- 17413104 TI - Isoflavonoid glucosides are deconjugated and absorbed in the small intestine of human subjects with ileostomies. AB - BACKGROUND: Although soy isoflavonoids have a number of health-promoting benefits, information concerning the sites of their absorption and metabolism in humans remains limited. Isoflavonoid absorption from the gut requires deconjugation of glucosides to aglycones. OBJECTIVE: The objective was to investigate the role of the small intestine in isoflavonoid absorption and metabolism in humans. DESIGN: Human subjects with fully functional gastrointestinal tracts (n = 6) and ileostomy subjects (n = 6) were fed a single soy meal containing 64.8 mg isoflavonoid aglycone equivalents (95% as glucosides). Metabolism of isoflavonoids in the upper gastrointestinal tract was examined by analyzing ileal effluent from ileostomy subjects, and absorption was assessed indirectly by quantifying isoflavonoids and several metabolites in 24-h urine pools. RESULTS: Chyme contained 36.7% of ingested isoflavonoid aglycone equivalents, primarily (95.8%) as aglycones. Qualitative profiles (x +/- SEM) of isoflavonoid excretion in urine (daidzein > glycitein > genistein) and the quantity of isoflavonoid equivalents were not significantly different between the control (18.4 +/- 2.2 mg) and ileostomy (13.5 +/- 3.2 mg) subjects. Dihydrodaidzein was present in the urine of all subjects, although the amount excreted by ileostomy subjects was less than that excreted by the control subjects. The percentage of producers and mean quantities of dihydrogenistein, equol, and O-desmethylangolensin in the urine of ileostomy subjects also were lower than those of control subjects. CONCLUSIONS: Ileostomy subjects efficiently deglycosylate isoflavonoid glucosides in the small intestine and appear to absorb aglycones with an efficiency comparable with that of control subjects. However, the production of microbial metabolites of isoflavonoids is limited in ileostomy subjects. PMID- 17413105 TI - Holo-transcobalamin is an indicator of vitamin B-12 absorption in healthy adults with adequate vitamin B-12 status. AB - BACKGROUND: It has been hypothesized that the response of holo-transcobalamin (holo-TC) to oral vitamin B-12 may be used to assess absorption. To develop a reliable clinical absorption test that uses holo-TC, it is necessary to determine the optimal timeline for vitamin B-12 administration and postdose assessment. OBJECTIVE: The objective of this study was to assess the magnitude and patterns of change in the postabsorption response of holo-TC to oral vitamin B-12. DESIGN: Adult (18-49 y) male and female participants (n = 21) with normal vitamin B-12 status were given three 9-mug doses of vitamin B-12 at 6-h intervals beginning early morning (baseline) on day 1. Blood was drawn at 17 timed intervals over the course of 3 d for the analysis of holo-TC and other indicators of vitamin B-12 status. RESULTS: Mean holo-TC increased significantly (P < 0.001) from baseline at 6 h (11%) and 24 h (50%). TC saturation increased significantly (P < 0.001) from baseline at 12.5 h (33%) and 24 h (50%). The mean cobalamin concentration changed significantly (P < 0.001) from baseline at 24 h (15%) and 48 h (14%). The ratio of holo-TC to cobalamin increased significantly (P < 0.001) at 24 h (32%). CONCLUSIONS: The greatest increase in holo-TC was observed 24 h after ingestion of three 9-mug doses of vitamin B-12. Our results indicate that a vitamin B-12 absorption test based on measurement of holo-TC after administration of three 9 mug doses of vitamin B-12 should run for 24 h. PMID- 17413106 TI - High prevalence of low dietary calcium, high phytate consumption, and vitamin D deficiency in healthy south Indians. AB - BACKGROUND: Data on the vitamin D status of the population in a tropical country such as India have seldom been documented. Vitamin D deficiency is presumed to be rare. OBJECTIVE: The objective was to document the dietary habits and concentrations of serum calcium, 25-hydroxyvitamin D [25(OH)D], and parathyroid hormone of Indian urban and rural populations. DESIGN: Healthy urban (n = 943) and rural (n = 205) subjects were studied for their dietary pattern and concentrations of serum calcium, phosphorus, alkaline phosphatase, 25(OH)D, and immunoreactive parathyroid hormone. RESULTS: The daily dietary calcium intake of both the urban and rural populations was low compared with the recommended dietary allowances issued by the Indian Council of Medical Research. Dietary calcium and phosphorous were significantly lower in rural adults than in urban adults (P < 0.0001). The dietary phytate-to-calcium ratio was higher in rural subjects than in urban subjects (P < 0.0001). The 25(OH)D concentrations of the rural subjects were higher than those of urban subjects (P < 0.001), both men and women. In the rural subjects, 25(OH)D-deficient (<20 ng/mL), -insufficient (20-30 ng/mL), and -sufficient (>30 ng/mL) states were observed in 44%, 39.5%, and 16.5% of the men and 70%, 29%, and 1% of the women, respectively. In the urban subjects, 25(OH)D-deficient, -insufficient, and -sufficient states were observed in 62%, 26%, and 12% of the men and 75%, 19%, and 6% of the women, respectively. CONCLUSIONS: Low dietary calcium intake and 25(OH)D concentrations were associated with deleterious effects on bone mineral homeostasis. Prospective longitudinal studies are required to assess the effect on bone mineral density, a surrogate marker for fracture risk and fracture rates. PMID- 17413107 TI - Magnesium intake and plasma concentrations of markers of systemic inflammation and endothelial dysfunction in women. AB - BACKGROUND: Relations between magnesium intake and systemic inflammation and endothelial dysfunction are not well established. OBJECTIVE: The aim of the present study was to examine whether and to what extent magnesium intake is related to inflammatory and endothelial markers. DESIGN: We conducted a cross sectional study of 657 women from the Nurses' Health Study cohort who were aged 43-69 y and free of cardiovascular disease, cancer, and diabetes mellitus when blood was drawn in 1989 and 1990. Plasma concentrations of C-reactive protein (CRP), interleukin 6 (IL-6), soluble tumor necrosis factor alpha receptor 2 (sTNF R2), E-selectin, soluble intercellular adhesion molecule 1 (sICAM-1), and soluble vascular cell adhesion molecule 1 (sVCAM-1) were measured. Estimates from 2 semiquantitative food-frequency questionnaires, administered in 1986 and 1990, were averaged to assess dietary intakes. RESULTS: In age-adjusted linear regression analyses, magnesium intake was inversely associated with plasma concentrations of CRP (P for linear trend = 0.003), E-selectin (P = 0.001), and sICAM-1 (P = 0.03). After further adjustment for physical activity, smoking status, alcohol use, postmenopausal hormone use, and body mass index, dietary magnesium intake remained inversely associated with CRP and E-selectin. Multivariate-adjusted geometric means for women in the highest quintile of dietary magnesium intake were 24% lower for CRP (1.70 +/- 0.18 compared with 1.30 +/- 0.10 mg/dL; P for trend = 0.03) and 14% lower for E-selectin (48.5 +/- 1.84 compared with 41.9 +/- 1.58 ng/mL; P for trend = 0.01) than those for women in the lowest quintile. CONCLUSION: Magnesium intake from diet is modestly and inversely associated with some but not all markers of systematic inflammation and endothelial dysfunction in apparently healthy women. PMID- 17413108 TI - Use of stable-isotope techniques to validate infant feeding practices reported by Bangladeshi women receiving breastfeeding counseling. AB - BACKGROUND: The World Health Organization recommends exclusive breastfeeding until age 6 mo. Studies relying on mothers' self-reported behaviors have shown that lactation counseling increases both the rate and duration of exclusive breastfeeding. OBJECTIVE: We aimed to validate reported infant feeding practices in rural Bangladesh; intakes of breast milk and nonbreast-milk water were measured by the dose-given-to-the mother deuterium dilution technique. DESIGN: Subjects were drawn from the large-scale Maternal and Infant Nutrition Interventions, Matlab, study of combined interventions to improve maternal and infant health, in which women were randomly assigned to receive either exclusive breastfeeding counseling or standard health care messages. Data on infant feeding practices were collected by questionnaire at monthly visits. Intakes of breast milk and nonbreast-milk water were measured in a subsample of 98 mother-infant pairs (mean infant age: 14.3 wk) and compared with questionnaire data reporting feeding practices. RESULTS: Seventy-five of the 98 subjects reported exclusive breastfeeding. Mean (+/-SD) breast milk intake was 884 +/- 163 mL/d in that group and 791 +/- 180 mL/d in the group reported as nonexclusively breastfed (P = 0.0267). Intakes of nonbreast-milk water were 40 +/- 80.6 and 166 +/- 214 mL/d (P < 0.0001), respectively. Objective cross-validation using deuterium dilution data showed good accuracy in reporting of feeding practices, although apparent misreporting was widely present in both groups. CONCLUSIONS: The dose-given-to the-mother deuterium dilution technique can be applied to validate reported feeding behaviors. Whereas this technique shows that the reports of feeding practices were accurate at the group level, it is not adequate to distinguish between feeding practices in individual infants. PMID- 17413109 TI - Subjective Global Nutritional Assessment for children. AB - BACKGROUND: Subjective Global Assessment (SGA), a method of nutritional assessment based on clinical judgment, has been widely used to assess the nutritional status of adults for both clinical and research purposes. OBJECTIVE: Foreseeing benefits of its use in children, we chose to adapt SGA and test its validity and reproducibility in the pediatric population. DESIGN: We prospectively evaluated the preoperative nutritional status of 175 children (aged 31 d to 17.9 y) having major thoracic or abdominal surgery with the use of Subjective Global Nutritional Assessment (SGNA) and commonly used objective measurements. Each child underwent nutritional assessment by 2 independent assessors, one performing measurements of anthropometrics and handgrip strength and one performing SGNA. To test interrater reproducibility, 78 children had SGNA performed by a third assessor. Occurrence of nutrition-associated complications was documented for 30 d postoperatively. RESULTS: SGNA successfully divided children into 3 groups (well nourished, moderately malnourished, severely malnourished) with different mean values for various anthropometric and biochemical measures (P < 0.05). Malnourished children had higher rates of infectious complications than did well-nourished children (P = 0.042). Postoperative length of stay was longer for malnourished children (8.2 +/- 10 d) than for well-nourished children (5.3 +/- 5.4 d) (P = 0.002). No objective nutritional measures showed association with outcomes, with the exception of serum albumin, which was not clinically predictive because mean concentrations were in the normal range irrespective of the presence or absence of complications. CONCLUSION: SGNA is a valid tool for assessing nutritional status in children and identifying those at higher risk of nutrition-associated complications and prolonged hospitalizations. PMID- 17413110 TI - Erythrocyte fatty acids and breast cancer risk: a case-control study in Shanghai, China. AB - BACKGROUND: The role of individual fatty acids in the development and progression of breast cancer is unclear. Although in vitro and animal experiments have supported an inverse association between intake of long chain n-3 fatty acids [primarily eicosapentaenoic acid (EPA) and docosahexaenoic acid] and breast cancer risk, findings from population studies are inconsistent. Recent studies have also shown associations between the ratio of saturated to monounsaturated fatty acids (SI) and breast cancer risk. The SI reflects the activity of several genes involved in lipid metabolism, including fatty acid synthase and steroyl coenzyme-A desaturase, that have been shown to be overexpressed in breast cancer. OBJECTIVE: The purpose of this analysis was to determine the association between erythrocyte fatty acid concentrations and breast cancer risk among women participating in a randomized trial of breast self-examination in Shanghai, China. DESIGN: We conducted a case-control study. Erythrocyte fatty acid concentrations were determined in specimens from 322 women with histologically confirmed breast cancer and 1030 frequency age-matched control women. RESULTS: We report a significant direct association among palmitic, gamma-linolenic, palmitoleic, and vaccenic acids and risk of breast cancer. Total n-3 fatty acids, EPA, and the SI for palmitic to palmitoleic acid were associated with significantly lower risk of breast cancer. CONCLUSION: Our results support a protective effect of n-3 fatty acids on breast cancer risk and provide additional evidence for the importance of evaluating the ratio of fatty acids when evaluating diet and breast cancer risk. PMID- 17413111 TI - A functional 19-base pair deletion polymorphism of dihydrofolate reductase (DHFR) and risk of breast cancer in multivitamin users. AB - BACKGROUND: Dihydrofolate reductase (DHFR) converts dihydrofolate (DHF) into tetrahydrofolate (THF) and plays an essential role in cell metabolism and cellular growth. Folic acid from multivitamins needs to be reduced by DHFR before it participates in cellular reactions. OBJECTIVES: We examined the relation of a 19-base pair (bp) deletion polymorphism of the DHFR gene with the risk of breast cancer by using data from the Long Island Breast Cancer Study Project, a population-based case-control study. We also investigated the transcriptional effect of this deletion polymorphism. DESIGN: Dietary data and habitual use of multivitamins were assessed from a modified Block food-frequency questionnaire (FFQ). Genotypes of DHFR were ascertained from 1062 case subjects and 1099 control subjects by allele-specific polymerase chain reaction. Unconditional logistic regression was used to estimate odds ratios (ORs) and 95% CIs. RESULT: Although the DHFR 19-bp deletion polymorphism was not associated with overall breast cancer risk, we observed a borderline significant additive interaction (P = 0.06) between the DHFR genotype and multivitamin use. The -19-bp allele was associated with greater breast cancer risk in multivitamin users (51.2% of the study population) with an OR of 1.26 (95% CI: 0.96, 1.66) and 1.52 (95% CI: 1.08, 2.13) for the +/- and -/- genotypes, respectively (P for trend = 0.02) than in multivatimin nonusers. A dose-dependent relation (P for trend < 0.001) between DHFR expression and the deletion genotype was observed. Compared with the subjects with the 19-bp +/+ genotype, subjects with the -/- genotype had 4.8-fold DHFR mRNA levels. CONCLUSIONS: The DHFR 19-bp deletion polymorphism affects the transcription of DHFR gene in humans. Multivitamin supplements may place a subgroup of women (ie, those with the -19-bp allele) at elevated risk of developing breast cancer. PMID- 17413112 TI - Plasma n-3 fatty acids and the risk of cognitive decline in older adults: the Atherosclerosis Risk in Communities Study. AB - BACKGROUND: Plasma fatty acids may affect the risk of cognitive decline in older adults. OBJECTIVES: We prospectively studied the association between plasma fatty acids and cognitive decline in adults aged 50-65 y at baseline and conducted a subgroup analysis. DESIGN: From 1987 through 1989, the Atherosclerosis Risk in Communities (ARIC) Study analyzed plasma fatty acids in cholesteryl esters and phospholipids in whites residing in Minneapolis, MN. From 1990 through 1992 and from 1996 through 1998, 3 neuropsychological tests in the domains of delayed word recall, psychomotor speed, and verbal fluency were administered. We selected cutoffs for statistically reliable cognitive decline in each of these domains and a measure of global cognitive change computed by principal-components analysis. Multivariate logistic regression was conducted. Focusing on n-3 highly unsaturated fatty acids (HUFAs), a subgroup analysis assessed differential association across potential effect modifiers implicated in oxidative stress and increased risk of neurodegenerative disease. RESULTS: In the 2251 study subjects, the risk of global cognitive decline increased with elevated palmitic acid in both fractions and with high arachidonic acid and low linoleic acid in cholesteryl esters. Higher n-3 HUFAs reduced the risk of decline in verbal fluency, particularly in hypertensive and dyslipidemic subjects. No significant findings were shown for psychomotor speed or delayed word recall. CONCLUSIONS: Promoting higher intakes of n-3 HUFAs in the diet of hypertensive and dyslipidemic persons may have substantial benefits in reducing their risk of cognitive decline in the area of verbal fluency. However, clinical trials are needed to confirm this finding. PMID- 17413113 TI - The contribution of plant foods to the vitamin A supply of lactating women in Vietnam: a randomized controlled trial. AB - BACKGROUND: More information is needed on the efficacy of carotenoids from plant foods in improving vitamin A status. OBJECTIVE: We aimed to quantify the efficacy of provitamin A-rich vegetables and fruit in improving vitamin A status. DESIGN: Breastfeeding women in 9 rural communes in Vietnam were randomly allocated to 1 of 4 groups: the vegetable group (n = 73), which ingested 5.6 mg beta-carotene/d from green leafy vegetables; the fruit group (n = 69), which ingested 4.8 mg beta carotene/d from orange or yellow fruit; the retinol-rich group (n = 70), which ingested 610 microg retinol/d from animal foods and 0.6 mg beta-carotene/d; and the control group (n = 68), which ingested 0.4 mg beta-carotene/d. Meals of groups 1, 2, and 4 contained <30 microg retinol/d. Lunch and dinner were provided 6 d/wk for 10 wk. RESULTS: Mean (95% CI) changes in serum retinol concentrations of the vegetable, fruit, retinol-rich, and control groups were 0.09 (0.03, 0.16), 0.13 (0.07, 0.19), 0.25 (0.17, 0.33), and 0.00 (-0.06, 0.06) micromol/L, respectively. Mean (95% CI) changes in breast-milk retinol concentrations were 0.15 (0.04, 0.27), 0.15 (0.02, 0.28), 0.48 (0.32, 0.64), and -0.06 (-0.21, 0.09) micromol/L, respectively. According to these findings, the equivalent of 1 microg retinol would be 12 microg beta-carotene (95% CI: 8, 22 microg) for fruit and 28 microg beta-carotene (17, 84 microg) for green leafy vegetables. Thus, apparent mean vitamin A activity of carotenoids in fruit and in leafy vegetables was 50% (95% CI: 27%, 75%) and 21% (7%, 35%), respectively, of that assumed. CONCLUSION: The bioavailability of carotenoids from vegetables and fruit is less than previously assumed. PMID- 17413114 TI - Body mass index and serum leptin concentration independently estimate percentage body fat in older adults. AB - BACKGROUND: Because serum concentrations of leptin, a hormone produced by adipocytes, can be relatively reliably and inexpensively measured, it may be considered complementary to, or even a substitute for, body mass index (BMI) as a measure of adiposity. OBJECTIVE: We examined the ability of BMI and leptin concentrations, separately and together, to estimate total percentage fat in older adults. DESIGN: Total percentage fat measured by dual-energy X-ray absorptiometry and fasting serum leptin concentrations were measured in 2911 well functioning 70-79-y-old participants (42% black, 51% women) in the Health, Aging, and Body Composition Study. RESULTS: Mean (+/-SD) total percentage fat was 29.2 +/-5.0% in men and 40.5 +/- 5.7% in women, and the geometric mean (+/-SD) serum leptin concentration was 5.6 +/- 2.5 ng/mL in the men and 16.4 +/- 2.3 ng/mL in the women. Among men, total percentage fat was strongly associated with both BMI (R(2) = 0.56) and leptin (R(2) = 0.57) in separate linear regression analyses and in a combined linear regression analysis (R(2) = 0.68). Similarly, among women, total percentage fat was associated with both BMI (R(2) = 0.65) and leptin (R(2) = 0.54) separately and in combination (model R(2) = 0.71). Independent relations of BMI and leptin with total percentage fat were also found among both black and white participants. With the population divided into quintiles according to percentage fat, BMI and serum leptin correctly classified 49% of men and 50% of women in the correct quintile. CONCLUSIONS: Among older adults, total percentage fat was better estimated by using both serum leptin concentrations and BMI than by using either alone. However, their performance does not suggest that they can substitute for more accurate measures. PMID- 17413115 TI - Community-level micronutrient fortification of a food supplement in India: a controlled trial in preschool children aged 36-66 mo. AB - BACKGROUND: Children participating in the Integrated Child Development Service (ICDS) in India have high rates of iron and vitamin A deficiency. OBJECTIVE: The objective was to assess the efficacy of a premix fortified with iron and vitamin A and added at the community level to prepared khichdi, a rice and dal mixture, in increasing iron and vitamin A stores and decreasing the prevalence of iron deficiency, anemia, and vitamin A deficiency. DESIGN: This cluster, randomized, double-blind, controlled trial was initiated in 30 Anganwadi centers (daycare centers) in West Bengal state, India. Children aged 36-66 mo (n = 516) attending village-based ICDS centers were randomly assigned to receive either a fortified or a nonfortified premix for 24 wk. Blood was drawn at 0 and 24 wk by venipuncture for the measurement of hemoglobin, serum ferritin, and serum retinol. RESULTS: The change in the hemoglobin concentration of anemic children was significantly different between fortified and nonfortified khichdi groups (P < 0.001). Prevalence rates of anemia, iron deficiency, and iron deficiency anemia were significantly lower after 24 wk in the fortified-khichdi group than in the nonfortified-khichdi group (P < 0.001). There were no significant differences in serum retinol concentrations or in the prevalence of vitamin A deficiency between the fortified- and nonfortified-khichdi groups. CONCLUSION: A premix fortified with iron, vitamin A, and folic acid and added to supplementary food at the community level can be effective at increasing iron stores and reducing the prevalence of iron deficiency and anemia. PMID- 17413116 TI - More acidic dietary acid-base load is associated with reduced calcaneal broadband ultrasound attenuation in women but not in men: results from the EPIC-Norfolk cohort study. AB - BACKGROUND: Dietary patterns that promote mild metabolic acidosis may have a negative effect on bone density. OBJECTIVE: We investigated the relation between a measure of dietary acid-base load, potential renal acid load (PRAL), and calcaneal broadband ultrasound attenuation (BUA) after adjustment for confounders and also compared the results with different estimates of acid-base load. DESIGN: A cross-sectional study was conducted in 14 563 men and women aged 42-82 y living in Norfolk, United Kingdom, in which measures of calcaneal BUA and dietary PRAL were estimated by using the European Prospective Investigation into Cancer and Nutrition Norfolk (EPIC-Norfolk) food-frequency questionnaire. RESULTS: A more acidic dietary intake (high PRAL) was significantly associated with lower calcaneal BUA in women but not in men; there was a difference of approximately 2% in BUA between the highest and lowest quintiles of PRAL, independent of age, body mass index, smoking habit, physical activity, diagnosed osteoporosis, and history of fracture, and (in women) hormone replacement therapy. No relation was observed between history of fracture or incident fracture and PRAL. Those with the greatest PRAL had higher intakes of meat, fish, eggs, and cereal and cereal products and lower intakes of fruit and vegetables, tea, and coffee. CONCLUSION: PRAL was inversely associated with bone ultrasound measures in women, but the magnitude of the association was relatively small compared with other known risk factors. Further longitudinal studies are required to establish whether, in the long term, these small effects are important in overall fracture risk in populations. PMID- 17413117 TI - Fish consumption, n-3 fatty acids, and subsequent 5-y cognitive decline in elderly men: the Zutphen Elderly Study. AB - BACKGROUND: Indications have been seen of a protective effect of fish consumption and the intake of n-3 fatty acids on cognitive decline. However, studies are scarce and results inconsistent. OBJECTIVE: The objective of the study was to examine the associations between fish consumption, the intake of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish and other foods, and subsequent 5-y cognitive decline. DESIGN: Data on fish consumption of 210 participants in the Zutphen Elderly Study, who were aged 70-89 y in 1990, and data on cognitive functioning collected in 1990 and 1995 were used in the study. The intake of EPA and DHA (EPA+DHA) was calculated for each participant. Multivariate linear regression analysis with multiple adjustments was used to assess associations. RESULTS: Fish consumers had significantly (P = 0.01) less 5-y subsequent cognitive decline than did nonconsumers. A linear trend was observed for the relation between the intake of EPA+DHA and cognitive decline (P = 0.01). An average difference of approximately 380 mg/d in EPA+DHA intake was associated with a 1.1-point difference in cognitive decline (P = 0.01). CONCLUSIONS: A moderate intake of EPA+DHA may postpone cognitive decline in elderly men. Results from other studies are needed before definite conclusions about this association can be drawn. PMID- 17413118 TI - Soy isoflavones lower serum total and LDL cholesterol in humans: a meta-analysis of 11 randomized controlled trials. AB - BACKGROUND: Clinical trials have reported the cholesterol-lowering effects of soy protein intake, but the components responsible are not known. OBJECTIVE: This meta-analysis was primarily conducted to evaluate the precise effects of soy isoflavones on lipid profiles. The effects of soy protein that contains enriched and depleted isoflavones were also examined. DESIGN: PUBMED was searched for English-language reports of randomized controlled trials published from 1990 to 2006 that described the effects of soy protein intake in humans. Eleven studies were selected for the meta-analysis. RESULTS: Soy isoflavones significantly decreased serum total cholesterol by 0.10 mmol/L (3.9 mg/dL or 1.77%; P = 0.02) and LDL cholesterol by 0.13 mmol/L (5.0 mg/dL or 3.58%; P < 0.0001); no significant changes in HDL cholesterol and triacylglycerol were found. Isoflavone depleted soy protein significantly decreased LDL cholesterol by 0.10 mmol/L (3.9 mg/dL or 2.77%; P = 0.03). Soy protein that contained enriched isoflavones significantly decreased LDL cholesterol by 0.18 mmol/L (7.0 mg/dL or 4.98%; P < 0.0001) and significantly increased HDL cholesterol by 0.04 mmol/L (1.6 mg/dL or 3.00%; P = 0.05). The reductions in LDL cholesterol were larger in the hypercholesterolemic subcategory than in the normocholesterolemic subcategory, but no significant linear correlations were observed between reductions and the starting values. No significant linear correlations were found between reductions in LDL cholesterol and soy protein ingestion or isoflavone intakes. CONCLUSIONS: Soy isoflavones significantly reduced serum total and LDL cholesterol but did not change HDL cholesterol and triacylglycerol. Soy protein that contained enriched or depleted isoflavones also significantly improved lipid profiles. Reductions in LDL cholesterol were larger in hypercholesterolemic than in normocholesterolemic subjects. PMID- 17413119 TI - Effects of soluble fiber (Plantago ovata husk) on plasma lipids, lipoproteins, and apolipoproteins in men with ischemic heart disease. AB - BACKGROUND: New dietary strategies to reduce cardiovascular disease (CVD) risk include the addition of fiber to the diet. The effect of soluble-fiber consumption derived from Plantago ovata husk on lipid risk factors in patients with CVD is unknown. OBJECTIVE: We compared the effects of soluble fiber (P. ovata husk) with those of insoluble fiber (P. ovata seeds) on plasma lipid, lipoprotein, and apolipoprotein (apo) concentrations within a CVD secondary prevention program. DESIGN: In a randomized, crossover, controlled, single-blind design, 28 men with CVD (myocardial infarction or stable angina) and an LDL cholesterol concentration or = A2 (median, 11.2; range, 5.4 - 51.7 ng/ml; normal median, 1.1; range, 0-2.3 ng/ml; p = 0.004). CONCLUSIONS: We have shown that elevated levels of pepsin, a biomarker of gastric aspiration, are consistently identified in the BAL of lung allografts. The highest levels were seen in patients with > or = grade A2 acute rejection. This provides further evidence supporting the possible role of aspiration in the development of overall allograft injury. PMID- 17413127 TI - Absence of T cells confers increased pulmonary arterial hypertension and vascular remodeling. AB - RATIONALE: Severe pulmonary arterial hypertension (SPH) is a frequently lethal condition characterized by pulmonary vascular remodeling and right heart strain or failure. SPH is also often associated with autoimmune and collagen vascular disorders. OBJECTIVES: To study the effects of T cells on the development of experimental SPH. METHODS: Athymic nude rats lacking T cells were treated with a single subcutaneous injection of vascular endothelial growth factor (VEGF) receptor blocker SU5416 (20 mg/kg) to induce pulmonary vascular endothelial cell apoptosis. Immunohistochemical analysis and IL-4 levels of the lung tissue were performed. Cell death and proliferation were assessed by Western blot and immunohistochemistry. MEASUREMENTS AND MAIN RESULTS: In contrast to SU5416 treated euthymic rats that develop SPH only in combination with chronic hypoxia, athymic nude rats developed SPH and vascular remodeling (similar to clinical SPH) at normoxic conditions as demonstrated by measurements of pulmonary artery pressure and right ventricle hypertrophy. Pulmonary arterioles became occluded with proliferating endothelial cells and were surrounded by mast cells, B cells, and macrophages. IL-4, proliferating cell nuclear antigen, and collagen type I levels were markedly increased in SU5416-treated athymic rat lungs. Antibody deposition was noted along the vascular endothelium in rats with SPH. Finally, protection from SPH was conferred by immune challenge with spleen cells from euthymic nude rats. CONCLUSIONS: These studies demonstrate the importance of a complete, intact immune system in protecting against pulmonary angioproliferation in this new model of SPH as well as the importance of intact VEGF receptor signaling for lung endothelial cell homeostasis. PMID- 17413128 TI - Diaphragm muscle fiber dysfunction in chronic obstructive pulmonary disease: toward a pathophysiological concept. AB - Inspiratory muscle weakness in patients with chronic obstructive pulmonary disease (COPD) is of major clinical relevance; maximum inspiratory pressure generation is an independent determinant of survival in severe COPD. Traditionally, inspiratory muscle weakness has been ascribed to hyperinflation induced diaphragm shortening. However, more recently, invasive evaluation of diaphragm contractile function, structure, and biochemistry demonstrated that cellular and molecular alterations occur, of which several can be considered of pathologic nature. Although the fiber-type shift toward oxidative type I fibers in COPD diaphragm is regarded as beneficial, rendering the overloaded diaphragm more resistant to fatigue, the reduction of diaphragm fiber force generation in vitro likely contributes to diaphragm weakness. The reduced diaphragm force generation at single-fiber level is associated with loss of myosin content. Moreover, the diaphragm in COPD is exposed to oxidative stress and sarcomeric injury. The current Pulmonary Perspective postulates that the oxidative stress and sarcomeric injury activate proteolytic machinery, leading to contractile protein wasting and, consequently, loss of force-generating capacity of diaphragm fibers in patients with COPD. Interestingly, several of these presumed pathologic alterations are already present early in the course of the disease (GOLD I/II), although these patients do not appear to be limited in their daily-life activities. Therefore, investigating in vivo diaphragm function in mild to moderate COPD should be the focus of future research. Treatment of diaphragm dysfunction in COPD is complex because its etiology is unclear, but recent findings show promise for the use of proteasome inhibitors in syndromes associated with muscle wasting, such as the diaphragm in COPD. PMID- 17413129 TI - The grand old men. PMID- 17413130 TI - California nursing facility quality and union environments. AB - OBJECTIVE: Many studies have examined quality effects of nursing facility (NF) staffing, but few have examined effects of unionization. Concerned with possible effects of unionization on quality, we analyzed unionization and local market climate of unionization, predicting both complaints (reflecting either quality problems or better monitoring and advocacy) and the substantiation of serious complaints (indicating major quality problems). METHOD: Data were analyzed on California freestanding NFs in 1999 (N = 1,155). OLS regression was employed to predict both quality complaints and serious violations, the latter both controlling and not controlling for numbers of complaints. RESULTS: Unionized NFs showed more complaints than did non-unionized NFs. Non-unionized NFs had more serious violations, particularly when the proportion of other county facilities unionized was higher. DISCUSSION: These findings suggest that unionization enhances problem reporting while, especially in stronger union environments, reducing the incidence of serious quality violations. PMID- 17413131 TI - Stride width discriminates gait of side-fallers compared to other-directed fallers during overground walking. AB - PURPOSE: The purpose was to identify differences in gait characteristics between older fallers with a tendency to fall sideways compared to those who do not fall to the side. METHOD: The authors conducted a prospective, case control study of ambulatory adults older than 70 residing in retirement communities. Measurements included spatial and temporal gait parameters and prospective fall surveillance. RESULTS: In all, 29 participants fell to the side, and 64 fell in other directions (forward, backward, straight down); 46 participants experienced no falls. Side-fallers exhibited narrower stride widths compared to other-directed fallers, and stepwise and discriminant analysis correctly classified 67% of side fallers and other-directed fallers using only stride width. DISCUSSION: This study suggests that side-fallers, who have narrower stride widths compared to those who fall in other directions, may not be adapting their gait to compensate for lateral instability. More research is needed to determine whether narrow gait contributes to unstable walking patterns. PMID- 17413132 TI - Risk factors for nursing home placement in older adults with and without dementia. AB - OBJECTIVE: The authors explored the effect of predisposing, enabling, and need characteristics on risk of nursing home placement (NHP) in participants with and without dementia. METHOD: Participants were 1,943 newly enrolled Medicare/Medicaid beneficiaries in Florida who were evaluated for health services by the Department of Elder Affairs. They were at least 65 years of age during fiscal year 1998-1999 and were followed for up to 4 years. RESULTS: Cox proportional hazard regressions indicated that baseline characteristics including older age, White race, diabetes, incontinence, stroke, and difficulties with instrumental activities increased risk of NHP in those without dementia. Caregiver, widowed status, arthritis, and heart disease reduced the risk. Only age, race, and marital status were predictive among participants with dementia. DISCUSSION: Dementia plays a central role in risk of NHP. Understanding risk factors for NHP in this relatively vulnerable population has important policy implications. PMID- 17413133 TI - Sensory impairment, use of community support services, and quality of life in aged care clients. AB - OBJECTIVE: To assess influences from visual or hearing impairment on use of community support services and health-related quality of life in aged care clients. METHOD: The authors sampled 284 frail elderly individuals presenting for assessment in Sydney, Australia. Moderate to severe visual impairment was defined as visual acuity <20/80 (better eye), and moderate to severe hearing loss as hearing thresholds >40 decibels (better ear). Community support services included home-delivered meals, home help, and community nurse visits. RESULTS: After adjusting for age, sex, and two or more comorbid conditions, moderate to severe visual impairment, but not moderate to severe hearing loss, was significantly associated with increased use of community services (adjusted odds ratio 2.8, 95% confidence interval = 1.0-7.8). CONCLUSION: Moderate to severe visual impairment was associated with an increased likelihood of community service utilization in this aged care client sample. PMID- 17413134 TI - Physical activity and the risk of dementia in oldest old. AB - OBJECTIVE: This study evaluated the protective role of physical activity (PA) against cognitive impairment (CI) in the oldest old (age >/= 85). METHOD: Prospective data on 66 optimally healthy, oldest old adults (mean age 88.5) were analyzed using survival analysis. RESULTS: In all, 12 men and 11 women reported exercising > 4 hours per week, and 38 participants developed CI (mean onset age 93; mean follow-up 4.7 years). The effect of exercise was modified by gender. In more active women (> 4 hours/week), the risk of CI was reduced by 88% (95% confidence interval 0.03, 0.41) compared to those less active. Less active women had 2 times the incidence rate of CI compared to less active men and almost 5 times the rate compared to active women. DISCUSSION: This study demonstrates the beneficial effects of exercise on healthy brain aging even in the oldest old and emphasizes the importance of increasing PA in older women. PMID- 17413135 TI - Cognitive impairment, psychiatric disorders, and problematic behaviors in a tribal nursing home. AB - OBJECTIVE: Residents' cognitive, psychiatric, and behavioral statuses were examined as part of a larger study of care in a nursing home (NH) owned and operated by a Northern Plains American Indian tribe. METHOD: Reviews of 45 medical records and semistructured interviews with 36 staff were completed. RESULTS: Creekside residents had considerable psychiatric and behavioral morbidity. High prevalences of non-Alzheimer's disease dementia, cognitive impairment, anxious symptomatology, and resistance to care were met with psychopharmacotherapy, reorientation, and informal techniques for behavior management. Significant depressive, anxious, psychotic, and behavioral symptoms remained. Staff interpretations of resident problems consisted of an ethnopsychological schema emphasizing resident loneliness, grumpiness, and propensity to "fight" rather than formal psychiatric nosology. DISCUSSION: Tribal NH residents were likely underdiagnosed for dementia and anxiety. Residual behavioral and psychiatric symptomatology suggest room for improvement in the NH's behavioral management regimen. Need for greater attention to conceptual, diagnostic, clinical, and documentation processes in the NH setting is noted. PMID- 17413136 TI - Premature graying, balding, and low bone mineral density in older women and men: the Rancho Bernardo study. AB - OBJECTIVE: The authors report the association of balding or graying with BMD in older adults. METHOD: BMD was measured at the spine, hip, and total body in 1,207 participants. Of these, 508 women and 380 men responded to a 1986 survey about balding patterns; in 1994, all participants answered questions about graying. RESULTS: Among men, 10.7% reported graying, and 51.1%, balding; 9.9% of women reported graying, and 9.5%, balding. Models were adjusted for age, body mass index, alcohol consumption, smoking, exercise, calcium supplements, diuretics, glucocorticoids, thyroid hormone, and estrogen. CONCLUSION: Graying was not significantly associated with BMD in either group. Balding men averaged 5% lower total body BMD (p /= 12 years were predictors of friendship loss. Higher educated African American women were at greater risk of non-spousal family loss. DISCUSSION: Future research should examine bereavement burden and identify health outcomes of multiple losses. PMID- 17413139 TI - Well-being and family support among elderly rural Mexicans in the context of migration to the United States. AB - OBJECTIVE: To explore perceptions of well-being, family support, and economic resources in relation to level of contact with migration to the U.S. among a sample of elderly males from rural Mexico. METHOD: The snowballing technique was used to obtain a sample of 372 participants. Four groups were created according to the level of contact with migration among older adults and their children. RESULTS: Greater level of contact with migration was associated with a higher likelihood that an older adult was literate, married or living with someone, self employed, and retired or pensioned. In addition, greater level of contact with migration to the U.S. was associated with a higher level of perceived well-being, family support, and economic security. DISCUSSION: Elderly, rural Mexican men with a greater degree of contact with migration to the U.S. seem to have more security and well-being in their old age. PMID- 17413140 TI - Screening for Chlamydia trachomatis. PMID- 17413141 TI - Coeliac disease in primary care. PMID- 17413142 TI - The 2006 WHO child growth standards. PMID- 17413143 TI - Intimate partner violence. PMID- 17413144 TI - Hospital acquired infection. PMID- 17413145 TI - UK doctors are already put off by changes in training. PMID- 17413146 TI - Let's open whole process of cost effective modelling. PMID- 17413147 TI - Improvement targets for C difficile must be valid. PMID- 17413149 TI - Total thyroidectomy is best operation for thyrotoxicosis. PMID- 17413152 TI - Appoint someone to make medical decisions if you are incapacitated, BMA says. PMID- 17413153 TI - Hewitt accuses some Muslim GPs of betraying confidentiality. PMID- 17413154 TI - Experts recommend circumcision to combat male HIV infections in Africa. PMID- 17413155 TI - Electronic prescribing needed in hospitals to monitor use of antibiotics. PMID- 17413156 TI - German doctors protest about firm offering fetal sex tests in early pregnancy. PMID- 17413158 TI - Safety agency makes better use of anticoagulants its top priority. PMID- 17413159 TI - Congress too prescriptive in saying how president's AIDS fund should be spent, report says. PMID- 17413163 TI - Virtual teaching offers practitioners new style of radiotherapy training. PMID- 17413170 TI - Duplicate publication: a bitter dispute. PMID- 17413171 TI - Life saving treatment or giant experiment? PMID- 17413172 TI - The royal colleges must up their game--or die. PMID- 17413173 TI - Screening programmes for chlamydial infection: when will we ever learn? PMID- 17413174 TI - Managing the menopause. PMID- 17413175 TI - Type 1 diabetes and pregnancy. PMID- 17413176 TI - Intermittent claudication. PMID- 17413182 TI - The role of somatostatin and dopamine receptors as molecular targets for the treatment of patients with pituitary adenomas. PMID- 17413183 TI - Novel insights in dopamine receptor physiology. AB - The dopaminergic system has a pivotal role in the central nervous system but also plays important roles in the periphery, mainly in the endocrine system. Dopamine exerts its functions via five different receptors, named D(1)-D(5), belonging to the category of G protein coupled membrane receptors. Dopamine receptors are heterogeneously expressed in different cells, tissues and organs, where they stimulate or inhibit different functions, including neurotransmission and hormone synthesis and secretion. In particular, the dopamineric system has a pivotal role in the physiological regulation of the hypothalamus-pituitary-adrenal axis. Recent data have demonstrated the expression and function of dopamine receptors not only in endocrine organs but also in endocrine tumors, mainly those belonging to the hypothalamus-pituitary-adrenal axis, and also in the so-called 'neuroendocrine' tumors. These data confirm the important role of the dopaminergic system in this endocrine axis, as well as in the neuroendocrine system. This review summarizes the main structural and functional characteristics of dopamine receptors, emphasizing the most recent novelties, and focused on the physiological and pathological regulation of the hypothalamus-pituitary-adrenal axis by the dopaminergic system. In addition, the recent findings on the relationship between dopamine receptors and neuroendocrine tumors are summarized. PMID- 17413184 TI - Novel chimeric somatostatin analogs: facts and perspectives. AB - Dopamine and somatostatin receptor agonists inhibit hormone secretion by normal pituitary cells and pituitary adenomas. Indeed, initially several dopaminergic drugs, and lately somatostatin analogs, have been developed for the treatment of pituitary adenomas. Recently, it has been demonstrated that subtypes of somatostatin and dopamine receptors may form homo- and hetero-dimers at the membrane level, as part of their normal trafficking and function. Interestingly, a specific ligand for a given receptor may influence the activity of an apparently unrelated receptor, and the association between the two different receptors could be induced by addition of either dopamine or somatostatin. The new properties of these families of G-protein coupled receptors (GPCRs) offer a potential explanation for the apparent conflicting results observed both in vivo and in vitro in human cell systems treated with the presently available analogs. Moreover, this observation not only increases the possibilities of modulating the activities of these receptors, but also raises new questions on the role of associations of specific receptors in the control of cell functions. In fact, results from preclinical studies have shown that receptor activation may not only trigger different intracellular signaling pathways, but also induce a distinct response depending upon the specific cell type. Recently, a number of new interesting compounds (subtype selective analogs and antagonists, as well as bi specific and hybrid somatostatin/dopamine compounds) have been developed. The effects of these new molecules have been explored in few animal and human cell lines and primary cultures from human tumors, revealing a heterogeneous, but broader, profile of activities. Further studies are certainly needed to fully elucidate the complex interplay between the GPCRs and consequent biological effects, to identify suitable therapies for controlling hormonal secretion of pituitary tumors. However, these recent observations form the basis for the application of new interesting strategies for the treatment of not only pituitary tumors but also other human malignancies. PMID- 17413185 TI - Control of pituitary adenoma cell proliferation by somatostatin analogs, dopamine agonists and novel chimeric compounds. AB - The antisecretory effects of somatostatin (SRIH) and its analogs are widely recognized and provide the basis for treatment of hormonal hypersecretion in patients with pituitary adenomas, especially in the settings of acromegaly. Dopamine (DA) agonists have also been used for medical treatment of prolactin and/or GH hypersecretion, and recent evidence points to an even greater antisecretory effect for a chimeric molecule, having high affinity for both SRIH and DA receptors. Evidence for an antiproliferative effect of these compounds has also been provided. This review focuses on the antiproliferative effects of SRIH and its analogs, of DA and chimeric compounds on pituitary adenomas, and on the clinical consequences on tumor volume of pharmacological treatment of pituitary adenomas with these drugs. PMID- 17413186 TI - Novel insights in somatostatin receptor physiology. AB - The experimental data reviewed in the present paper deal with the molecular events underlying the agonist-dependent regulation of the distinct somatostatin receptor subtypes and may suggest important clues about the clinical use of somatostatin analogs with different pattern of receptor specificity for the in vivo targeting of tumoral somatostatin receptors. Somatostatin receptor subtypes are characterized by differential beta-arrestin trafficking and endosomal sorting upon agonist binding due, at least in part, to the differences in their C terminal tails. Moreover, the subcellular expression pattern of somatostatin receptor subtypes and their activity in response to agonist treatment are affected by intracellular complements, such as proteins involved in intracellular vesicle trafficking. Different somatostatin analogs may induce distinct conformations of the receptor/ligand complex, preferentially coupled to either receptor signaling or receptor endocytosis. PMID- 17413187 TI - Preclinical and clinical experiences with the role of dopamine receptors in the treatment of pituitary adenomas. AB - Pituitary tumors can cause symptoms of mass effect and hormonal hypersecretion that can be reversed with surgical resection or debulking of the adenoma, radiotherapy, or medical treatment. Medical treatment is the primary choice for prolactinomas because dopamine agonists are very effective in the treatment of these tumors, with rates of control (tumor size reduction and hormone suppression) as high as 80-90% for microprolactinomas and 60-75% for macroprolactinomas. The function of dopamine receptors in other histotypes of pituitary adenoma is still debated. However, new insights into receptor physiology and the introduction of new clinically available, as well as experimental, compounds have reopened a potential role of dopaminergic drugs in the medical treatment of pituitary tumors. The differences between the effectiveness and the resistance to different dopaminergic agents, the new challenging results from clinical and experimental studies, as well as the future of dopamine agonists in the therapy of pituitary tumors are discussed. PMID- 17413188 TI - Preclinical and clinical experiences with the role of somatostatin receptors in the treatment of pituitary adenomas. AB - The patho-physiological role of somatostatin receptor subtypes (sst) in neuro endocrine diseases has gained enhanced scientific interest in the past few years. The development of novel somatotropin-release inhibiting factor analogs, both sst specific and universal ligands, seem promising as a tool to further increase fundamental insights in sst function. Eventually, this research should result in novel medical therapeutic opportunities in patients suffering from neuro endocrine diseases. In the present review, the functional role of sst in all types of pituitary adenomas, based on recent preclinical and clinical studies, is being discussed. PMID- 17413189 TI - Diagnostic imaging of dopamine receptors in pituitary adenomas. AB - Dopamine D2 receptor scintigraphy of pituitary adenomas is feasible by single photon emission computed tomography using (123)I-S-(-)-N-[(1-ethyl-2 pyrrolidinyl)methyl]-2-hydroxy-3-iodo-6-methoxybenzamide ((123)I-IBZM) and (123)I epidepride. (123)I-epidepride is generally superior to (123)I-IBZM for the visualization of D2 receptors on pituitary macroadenomas. However, (123)I-IBZM and (123)I-epidepride scintigraphy are generally not useful to predict the response to dopaminergic treatment in pituitary tumour patients. These techniques might allow discrimination of non-functioning pituitary macroadenomas from other non-tumour pathologies in the sellar region. Dopamine D2 receptors on pituitary tumours can also be studied using positron emission tomography with (11)C-N raclopride and (11)C-N-methylspiperone. PMID- 17413190 TI - Combined therapy of somatostatin analogues and dopamine agonists in the treatment of pituitary tumours. AB - Pituitary tumours express both somatostatin and dopamine receptors. Medical treatment with somatostatin analogues is a cornerstone of GH- and TSH-secreting tumours, while treatment with dopamine agonists is a cornerstone of prolactin secreting tumours. Dopamine agonists have also demonstrated some efficacy in patients with GH- and TSH-secreting adenomas. Neither ACTH-secreting nor clinically non-functioning tumours have a well-established medical treatment. Nevertheless, some recent results have indicated a potential usefulness of the dopamine agonist cabergoline in patients with pituitary-dependent Cushing's disease. Combination treatment with both somatostatin analogues and dopamine agonists has been poorly investigated. Some studies conducted in small series have documented an additive effect of both drugs in patients with GH-secreting adenomas. Of mention is that none of the studies were randomised and cross sectional so that the results should be confirmed by other well-designed studies. No data are available in other pituitary tumour histotypes. Preliminary observations in patients with clinically non-functioning adenomas are very promising. PMID- 17413191 TI - Integrated management of neonatal and childhood illnesses: a new hope for child survival. PMID- 17413192 TI - The pediatric management of snakebite the national protocol. PMID- 17413193 TI - Low birth weight and insulin resistance in mid and late childhood. AB - OBJECTIVE: Low birth weight is associated with adult insulin resistance and diabetes. We conducted this study to correlate low birth weight with insulin resistance in mid and late childhood. METHODS: Children whose birth weight records were available were successively enrolled from middle and low socioeconomic status urban schools in western India. 600 children in age groups 5 16 years were screened for availability of recorded birth weight in six schools. Detailed birth records were available for 158 children born full term. Parents of 134 (84.8%) agreed to participate in the study after informed consent. These children were evaluated for various anthropometric indices and fasting blood was obtained for determination of glucose and insulin levels. Insulin resistance was determined using homeostasis model assessment (HOMA) and HOMA-2 formula. Comparative, univariate and multivariate statistical analyses were performed. RESULTS: The mean age of the children was 10.0 +/- 2.4 years. Maternal diabetes was present in 3 (2.2%). Mean weight at birth was 2.84 +/- 0.61 kg and low birth weight (<2.5 kg) was in 49 children (36.6%). There was no significant statistical difference in current height, weight, body mass index, waist, hip, waist hip ratio, mid upper arm circumference, and systolic and diastolic blood pressure in children born with low or normal birth weight (P >0.10). In low birth weight as compared to normal birthweight children mean fasting blood glucose (80.1 +/- 16.1 vs. 70.1 +/- 14.8 mg/dL; P = 0.042), median (interquartile range) fasting insulin levels (10. 18, 6.08-18.54 vs. 2.12, 0.02-7.45 microU/mL; Mann Whitney U test, p<0.0001), and HOMA-derived insulin resistance (1.88, 1.06-4.52, vs. 0.35, 0.02 1.52, P < 0.0001) were significantly greater. There was a significant negative correlation of birth weight with fasting insulin levels after multifactorial adjustments (partial correlation coefficient r = -0.436, P <0.001). Multivariate conditional logistic regression analysis revealed that birthweight was significant determinant of fasting insulin levels after adjusting for age, gender, body mass index, waist and waist hip ratio (odds ratio 3.82, 95% confidence intervals 1.16-12.63, P = 0.028). CONCLUSIONS: Children born with low birth weight have significantly greater fasting glucose, fasting insulin levels and insulin resistance at mid and late childhood. PMID- 17413194 TI - IAP growth monitoring guidelines for children from birth to 18 years. PMID- 17413195 TI - Efficacy of breast milk gastric lavage in preterm neonates. AB - OBJECTIVE: To evaluate effects of gastric lavage with mother's milk starting 4 hours after birth, in hospitalized preterm newborns otherwise on exclusive parenteral fluids. METHOD: STUDY DESIGN: Randomized controlled trial. Sick preterm babies were assigned to receive in addition to parenteral fluids, either gastric lavage with mother's milk within 4 hours of birth and subsequently every 3 hours till tolerance of nutritive enteral feeds (intervention or BML group, n = 40), or remain nil per orally till tolerance of nutritive enteral feeds (control or NPO group, n = 40). The main outcome was the mean number of days of parenteral fluids till successful tolerance of nutritive enteral feeds. They were also evaluated for mean duration of hospital stay, development of new complications, and mortality. RESULT: Despite sicker babies in the BML group at baseline, the mean duration of exclusive parenteral fluid was significantly less (P = 0.003) in BML (3.9 +/- 1.5 days as compared to 5.4 +/- 2.6 days in NPO). In the NPO group 60% of the babies stayed longer than 3 weeks in hospital compared to only 30% in the BML group. The risk of development of new complication after randomization was also significantly less in BML group [RR 0.61 (95% CI 0.40-0.95) (P=0.03)]. Incidence of sepsis was 44% less in BML group [30% in BML, 55% in NPO group; RR 0.58; 95% CI 0.35-0.97; P = 0.02]. On multivariate logistic regression, BML group, birth weight and absence of complication at the time of hospitalization were strong predictors of improved outcome. There was no difference in mortality between 2 groups. CONCLUSION: This study showed that early exposure to even small amounts of mother's milk in sick preterm neonates significantly reduced the days on parenteral fluids, risk of sepsis and the duration of hospital stay without any adverse effect. PMID- 17413196 TI - Nutritional status of affluent Indian school children: what and how much do we know? AB - OBJECTIVE: This paper reviews available literature on nutritional status of Indian school children 6-18 years from middle and high socio economic status (MHSES). METHODS: Literature search was conducted using Medline literature database search, followed by review of full length journal papers and unpublished materials such as research reports. RESULTS: Studies showed that anemia prevalence (hemoglobin concentration <120 g/L) ranged from 19 to 88% across five different cities in India. Other micronutrient deficiencies including, folate, riboflavin, niacin, vitamin C, vitamin A, and vitamin B12 were also present based on biochemical parameters in one study and clinical signs of deficiency in three other studies. Overweight and obesity were prevalent among 8.5-29.0% and 1.5-7.4% respectively among school children, as indicated by 11 studies. Predominant components in children's diet were cereals and pulses, followed by milk and milk products; the fruits and vegetables component was comparatively lower. CONCLUSION: Nutritional status of MHSES children in India needs attention especially with respect to the high prevalence of anemia, overweight and obesity. There are indications that micronutrient deficiencies exist, but sufficient data are lacking, in particular biochemical data. A current estimate, using well designed methodologies, of prevalence of micronutrient deficiencies and information on the etiology of anemia among children of MHSES groups would be valuable to help understand the nutritional status and extent of micronutrient malnutrition. PMID- 17413197 TI - Chlamydia sp. in hospitalised children with community acquired pneumonia. AB - Our aim was to document the prevalence of chlamydial infection in children less than five years of age with Community Acquired Pneumonia (CAP). Seventy three children, 1 month to 5 years of age, hospitalized with CAP were enrolled over a period of one year. Microimmunofluorescence (MIF) was done to detect IgM antibodies against Chlamydia sp. in sera of all patients; PCR was performed to detect C. pneumoniae DNA in nasopharyngeal aspirates. The prevalence of Chlamydia species infection in CAP in children < 5 years of age was 5.5% (4/73). Two cases were positive for C. trachomatis antibodies; one case was positive for C. pneumoniae antibodies and one case was positive for C. pneumoniae DNA. Chlamydia sp. have an important role in CAP in children < 5 years and for early diagnosis of infection, use of more than one method i.e. PCR and serology both is advisable. PMID- 17413198 TI - Cranial neuroimaging in infantile tremor syndrome (ITS). AB - Clinical, hematological and cranial neuroimaging findings of eight cases of infantile tremor syndrome are reported. All had coarse tremors, anemia, hyperpigmentation and delayed or regression of developmental milestones. Five patients had microcytic, hypochromic anemia, three had dimorphic anemia. CT scans of two cases and MRI scans of three cases showed cerebral atrophy. One of these two CT scans, in addition, showed a small hypodensity in right basal ganglia region. Two CT scans were normal. One MRI showed hyperintense signals in frontal and periventricular white matter on T2 weighted images. The changes described are non-specific and also seen in cases of malnutrition and viral infections of CNS. PMID- 17413201 TI - Fanconi-Bickel syndrome. AB - We present here the first case of Fanconi-Bickel syndrome, a rare type of glycogen storage disease, from India. A 17-month-old female child presented with severe growth retardation and abdominal distention. Clinical examination revealed a "doll-like" face, massive hepatomegaly, and rickets. Laboratory investigations confirmed severe hypophosphatemic rickets and proximal renal tubular dysfunction. Liver biopsy showed glycogen accumulation in the hepatocytes. PMID- 17413202 TI - Hepatic abscess in a preterm neonate. AB - We present a preterm (32 weeks; weight 1300 g) neonate, with solitary hepatic abscess. The causative organism was Klebsiella. The child presented with septicemia, hepatomegaly and abdominal lump. Radiological investigations were inconclusive. Surgical drainage and prolonged antibiotic treatment led to resolution of the hepatic abscess. PMID- 17413203 TI - Biotin responsive limb weakness. AB - We report acute onset quadriplegia in a ten year old boy associated with basal ganglia lesions due to biotin deficiency. Prolonged raw egg consumption was identified as predisposing factor for biotin deficiency. Treatment with biotin resulted in remarkable recovery. PMID- 17413204 TI - Vulval condylomata acuminata. PMID- 17413205 TI - Snake bite neurotoxicity: reversal after 84 hours. PMID- 17413206 TI - Cerebral blood flow in Sydenham's chorea. PMID- 17413207 TI - An outbreak of mumps meningoencephalitis in Sangli district. PMID- 17413208 TI - Severe hyperkalemia in nephrotic syndrome. PMID- 17413209 TI - Cu(Cu0.44Cr4.56)Ge2O12: a close-packed oxide with CuO4 tetrahedra. AB - The structure of copper(I,II) pentachromium(III) germanate, Cu(Cu(0.44)Cr(4.56))Ge(2)O(12), contains one Cu position (m2m), one Ge position (m) and three Cr positions (2/m, m and 2). The close-packed structure is described in terms of slabs of edge-sharing Cr(3+)O(6) octahedra and isolated CuO(4) and GeO(4) tetrahedra. These slabs are aligned parallel to the bc plane and are separated from each other by GeO(4) tetrahedra along a. The tetrahedral coordination observed for the Cu(+)/Cu(2+) ions represents an unusual feature of the structure. The Cr-O and Cu-O bond lengths are compared with literature data. PMID- 17413210 TI - A pseudo-merohedrally twinned rare-earth sulfate: K6[Ce(HSO4)2(SO4)4].H2O, a novel structure type. AB - A novel structure type of an acidic rare-earth sulfate, hexapotassium cerium dihydrogensulfate tetrasulfate monohydrate, is reported. The crystal is twinned, mimicking tetragonal symmetry. The Ce(IV) atom is nine-coordinate, connecting to one corner-sharing and four edge-sharing sulfate groups. One of the potassium ions is disordered over two general positions. The compound is unique as it contains rare-earth monomers, [Ce(HSO(4))(SO(4))(4)](5-). The structure is composed of these monomers, water molecules, discrete hydrogensulfate ions and potassium ions held together by ionic interactions. There are two types of alternating layers in the structure, with compositions [K(4)Ce(HSO(4))(SO(4))(4)](-) and [K(2)(HSO(4))(H(2)O)](+). PMID- 17413211 TI - LiMn3(SeO3)2(HSeO3)6. AB - The title compound, lithium trimanganese bis[trioxoselenate(IV)] hexakis[hydrogentrioxoselenate(IV)], is built up from a vertex-sharing network of distorted Mn(III)O(6) octahedra, SeO(3) and HSeO(3) pyramids and unusual Li(OH)(6) octahedra, resulting in a dense three-dimensional structure. Mn, Li and one Se atom have site symmetries of -1, -3, and 3, respectively. An O-H...O hydrogen bond helps to establish the crystal packing. PMID- 17413212 TI - Poly[mu2-trans-1,2-di-4-pyridylethylene-kappa2N:N'-mu2-sulfato-kappa2O:O' zinc(II)]. AB - The title compound, [Zn(SO(4))(C(12)H(10)N(2))](n), features a layered structure based on [Zn(SO(4))](n) spirals linked by 1,2-di-4-pyridylethylene (bpe) ligands, with the tetrahedral Zn and S atoms lying on twofold axes. The bpe ligands are centrosymmetric. The layers are linked by weak C-H...O interactions. PMID- 17413213 TI - Bis(2-amino-3,5-dibromo-4,6-dimethylpyridinium) hexabromidostannate(IV). AB - In the title compound, (C(7)H(9)Br(2)N(2))(2)[SnBr(6)], the cations and centrosymmetric anions are stacked in alternating layers that show no significant intermolecular interactions within each stack. Extensive cation...[SnBr(6)](2-) interactions are found, represented by short Br...Br interactions, along with different Br...HN (pyridine and amine) and weaker Br...HCH(2) hydrogen-bonding motifs. PMID- 17413214 TI - Diaquabis[4-(4H-1,2,4-triazol-4-yl)benzoato-kappa2O,O']cobalt(II), and the cadmium(II) and copper(II) analogues: new self-complementary hydrogen-bond donor/acceptor modules for designing hydrogen-bonded frameworks. AB - In the isostructural title complexes, [M(C(9)H(6)N(3)O(2))(2)(H(2)O)(2)] [M = Co(II), (I), Cd(II), (II), and Cu(II), (III); the metal centres reside on a twofold axis in the space group C2/c for (I) and (II)], the metal centres are surrounded by four O atoms from two O,O'-bidentate carboxylate groups and by two trans-coordinated aqua ligands, forming a distorted octahedral environment. The molecules possess four hydrogen-bond donor (two aqua ligands) and four hydrogen bond acceptor sites (two triazole groups), and aggregate by self-association, forming two-dimensional hydrogen-bonded frameworks [via O-H...N interactions; O...N = 2.749 (3)-2.872 (3) A]. The layers are parallel and are tightly packed with short interlayer distances of 4.93, 4.95 and 5.01 A for (I), (II) and (III), respectively. PMID- 17413215 TI - Chloridobis[(2-oxoazocan-1-yl)methyl]germanium(IV) trifluoromethanesulfonate. AB - In the title compound, [Ge(C(8)H(14)NO)(2)Cl]CF(3)SO(3), which is the first complex containing an eight-membered lactam (enantholactam) as ligand, the coordination polyhedron of the Ge(IV) atom is intermediate between trigonal bipyramidal and square-pyramidal. Quantum chemical calculations of the crystal structure indicate the absence of additional coordination bonding between the Ge(IV) atom and the trifluoromethanesulfonate anion. PMID- 17413216 TI - trans-Diaqua{5,5'-[(E,E)-pyridine-2,6-diylbis(methylidynenitrilo)]bis[pyrimidine 2,4(1H,3H)-dione]}zinc(II) nitrate hexafluorophosphate trihydrate. AB - The title compound, [Zn(C(15)H(11)N(7)O(4))(H(2)O)(2)](NO(3))(PF(6)).3H(2)O, contains a mononuclear zinc(II) complex. The Zn(2+) centre is seven-coordinated in a slightly distorted pentagonal-bipyramidal geometry, with the two water O atoms located in the apical positions, and the pyridine N atom, the two imine N atoms and two carbonyl O atoms of the uracil groups located in the equatorial plane. The charge is balanced by PF(6)(-) and NO(3)(-) anions. PMID- 17413217 TI - A new polymorph of a cobalt(III) Schiff base complex exhibits a one-dimensional C H...O hydrogen-bonded extended structure with helical 2 1 symmetry. AB - In a new polymorph of acetato(2,2'-{iminobis[(E)-propane-3,1 diylnitrilomethylidyne]}diphenolato)cobalt(III), [Co(C(20)H(23)N(3)O(2))(C(2)H(3)O(2))], in the space group P2(1)/c, the Co(III) ion is six-coordinate, with unequal Co-O(phenolate) distances that average 1.908 (12) A and more similar Co-N(imine) distances averaging 1.937 (4) A. The acetate ion occupies the sixth coordination site and forms an intramolecular hydrogen bond (H...O = 1.95 A) with the Co-bound NH group of the pentadentate chelate. The extended structure is a one-dimensional (aryl)C-H...O(carbonyl) hydrogen-bonded polymer with 2(1) (helical) symmetry and is thus distinct from the simple hydrogen-bonded stack of the P2(1)/n polymorph [Matsumoto, Imaizumi & Ohyoshi (1983). Polyhedron, 2, 137-139]. PMID- 17413218 TI - An unusual P3 1 2 framework for the Ni(4,4'-bipyridine)(H2O)4 chain. AB - catena-Poly[[[tetraaquanickel(II)]-mu-4,4'-bipyridine-kappa(2)N:N'] thiosulfate dihydrate], {[Ni(C(10)H(8)N(2))(H(2)O)(4)]S(2)O(3).2H(2)O}(n), (I), and catena poly[[[tetraaquanickel(II)]-mu-4,4'-bipyridine-kappa(2)N:N'] sulfate methanol solvate monohydrate], {[Ni(C(10)H(8)N(2))(H(2)O)(4)]SO(4).CH(4)O.H(2)O}(n), (II), are built up of {[Ni(4,4'-bipy)(H(2)O)(4)](2+)}(n) chains (4,4'-bipy is 4,4' bipyridine) interwoven in an unusual P3(1) fashion. Voids are filled by the corresponding counter-anions and solvate molecules, defining a complex three dimensional network surrounding them. In both structures, the cationic chains evolve around a set of twofold axes passing through the Ni(II) ions and bisecting the aromatic amines through their N (and their opposite C) atoms. PMID- 17413219 TI - Poly[[mu2-4,4'-bipyridine-di-mu3-sulfito-dizinc(II)] monohydrate]. AB - The title compound, {[Zn(2)(SO(3))(2)(C(10)H(8)N(2))].H(2)O}(n), is a two dimensional polymer built up of a [ZnSO(3)](n) chain evolving around a 2(1) axis and interconnected by a 4,4'-bipyridine spacer. The resulting two-dimensional structures are linked, in turn, by hydrogen bonding mediated by the solvent water molecule. The organic ligand lies on a centre of symmetry located on the mid point of the bond between the rings, while the solvent water molecule is halved by a twofold axis passing through the O atom. PMID- 17413220 TI - A new two-dimensional CdII coordination polymer constructed by pyrazine-2,3 dicarboxylate. AB - In the title compound, poly[mu(5)-pyrazine-2,3-dicarboxylato-cadmium(II)], [Cd(C(6)H(2)N(2)O(4))](n) or [Cd(pdc)](n), where pdc is the pyrazine-2,3 dicarboxylate anion, the Cd(II) atom is six-coordinated by five carboxylate O atoms and one N atom from five different pdc ligands in a distorted octahedral CdO(5)N coordination geometry. Two Cd(II) atoms are bridged by carboxylate groups of the pdc ligands to create a dimeric unit. The dimeric units are further connected by the pdc ligands to generate an interesting two-dimensional structure. PMID- 17413221 TI - A new two-dimensional supramolecule composed of mu-2,6-dimethylpyridine-3,5 dicarboxylato-kappa4O3,O3':O5,O5'-bis[chloro(1,10-phenanthroline kappa2N,N')zinc(II)]. AB - In the binuclear title molecule, [Zn(2)(C(9)H(7)NO(4))Cl(2)(C(12)H(8)N(2))(2)], the two metal centres are bridged by a 2,6-dimethylpyridine-3,5-dicarboxylate ligand. The binuclear unit is extended to form a two-dimensional supramolecular motif via pi-pi stacking interactions between neighbouring phenanthroline rings. PMID- 17413222 TI - Unexpected oxidation of a diphosphine by bis(1,3-diphenylpropane-1,3 dionato)cobalt(II), [Co(dbm)2]. AB - The reaction of bis(1,3-diphenylpropane-1,3-dionato)cobalt(II), [Co(dbm)(2)], with bis(diphenylphosphino)ethane (dppe) affords the coordination polymer catena poly[[bis(1,3-diphenylpropane-1,3-dionato-kappa(2)O,O')cobalt(II)]-mu ethylenebis(diphenylphosphine oxide)-kappa(2)O:O'], trans [Co(C(15)H(11)O(2))(2)(C(26)H(24)O(2)P(2))](n), as a result of oxidation of the diphosphine. The Co atom is octahedral, with a CoO(6) coordination sphere, and the chelating dbm ligands adopt a trans configuration. The Co atom also lies on a centre of inversion, with a further symmetry centre bisecting the bridging ethylenebis(diphenylphosphine oxide) ligand. PMID- 17413223 TI - (Croconato-kappa2O,O')bis(1,10-phenanthroline-kappa2N,N')cobalt(II), and the nickel(II) and copper(II) analogues. AB - The title complexes, [M(C5O5)(C12H8N2)2], with M = Co(II), Ni(II) and Cu(II), all lie across twofold rotation axes, around which two 1,10-phenanthroline ligands are arranged in a chiral propeller manner. The Co(II) and Ni(II) complexes are isostructural, with octahedral coordination geometry, while the local geometry of the Cu(II) complex is severely distorted from octahedral. PMID- 17413224 TI - Sodium pyridine-3-carboxylate. AB - The title compound, also known as sodium nicotinate, Na(+).C(6)H(4)NO(2)(-), consists of two unique Na atoms coordinated to two unique pyridine-3-carboxylate ligands through the N atoms and carboxylate groups. One Na atom and one pyridine 3-carboxylate ligand lie on a twofold axis. Extensive Na coordination results in a three-dimensional array comprising infinite NaO(2)CR chains linked by intrachain Na-N bonds. PMID- 17413225 TI - Hydrogen bonding and pi-pi stacking in the three-dimensional supramolecular complex bis(4,4'-bipyridinium) diaquadioxalatoferrate(II) bis(hydrogen oxalate). AB - The title compound, (C(10)H(10)N(2))(2)[Fe(C(2)O(4))(2)(H(2)O)(2)](C(2)HO(4))(2), appears to be a modular associate consisting of a complex anion containing bivalent Fe as the central atom, a bridging hydrogen oxalate anion and a diprotonated 4,4-bipyridine acting as the counter-cation. The Fe(II) ion in the complex anion occupies a position on a centre of inversion. Its coordination environment is formed by six O atoms from two bidentate oxalate ligands, forming a basal plane, and two water molecules approximately perpendicular to the plane, representing a distorted octahedral geometry. These three kinds of ions are connected by strong hydrogen bonds, with donor-acceptor distances (N...O and O...O) lying in the range 2.54-2.98 A, and pi-pi stacking interactions between the 4,4'-bipyridinium cations, thus forming a three-dimensional supramolecular structure. PMID- 17413226 TI - A twinned triclinic polymorph of dibromidotetrakis(tetrahydrofuran kappaO)magnesium(II). AB - The title compound, [MgBr(2)(C(4)H(8)O)(4)], forms crystals which appear to be monoclinic but are actually twinned triclinic. The current form is a new triclinic polymorph, with Z'= 2, in addition to the already known tetragonal polymorph. Although the geometric parameters of the two polymorphs agree well, their packing patterns are completely different. PMID- 17413227 TI - Aqua(2,2'-diamino-4,4'-bi-1,3-thiazole-kappa2N,N')(oxydiacetato kappa3O,O',O'')cobalt(II) trihydrate. AB - The title compound, [Co(C(4)H(4)O(5))(C(6)H(6)N(4)S(2))(H(2)O)].3H(2)O, displays a distorted octahedral coordination geometry. The tridentate oxydiacetate dianion chelates the Cu(II) atom in the meridional mode. In the crystal packing, hydrophilic and hydrophobic layers are arranged in an alternating manner. In addition, a three-dimensional hydrogen-bonding framework and pi-pi stacking are present. PMID- 17413228 TI - Isomeric Schiff bases related by dual imino-group reversals. AB - Two isomeric pairs of Schiff bases, N,N'-bis(2-methoxybenzylidene)-p phenylenediamine, C(22)H(20)N(2)O(2), (I), and 2,2'-dimethoxy-N,N-(p phenylenedimethylene)dianiline, C(22)H(20)N(2)O(2), (II), and (E,E)-1,4-bis(3 iodophenyl)-2,3-diazabuta-1,3-diene (alternative name: 3-iodobenzaldehyde azine), C(14)H(10)I(2)N(2), (III), and N,N'-bis(3-iodophenyl)ethylenediimine, C(14)H(10)I(2)N(2) [JAYFEV; Cho, Moore & Wilson (2005). Acta Cryst. E61, o3773 o3774], differ pairwise only in the orientation of their imino linkages and in all four individual cases occupy inversion centers in the crystal, yet all four compounds are found to assume unique packing arrangements. Compounds (I) and (II) differ substantially in molecular conformation, possessing angles between their ring planes of 12.10 (15) and 46.29 (9) degrees , respectively. Compound (III) and JAYFEV are similar to each other in conformation, with angles between their imino linkages and benzene rings of 11.57 (15) and 7.4 (3) degrees , respectively. The crystal structures are distinguished from each other by different packing motifs involving the functional groups. Intermolecular contacts between methoxy groups define an R(2)(2)(6) motif in (I) but a C(3) motif in (II). Intermolecular contacts are of the I...I type in (III), but they are of the N...I type in JAYFEV. PMID- 17413229 TI - Supramolecular hydrogen-bonding networks in bis(adeninium) phthalate phthalic acid 1.45-hydrate. AB - In the title compound, 2C(5)H(6)N(5)(+).C(8)H(4)O(4)(2-).C(8)H(6)O(4).1.45H(2)O, the asymmetric unit comprises two adeninium cations, two half phthalate anions with crystallographic C(2) symmetry, one neutral phthalic acid molecule, and one fully occupied and one partially occupied site (0.45) for water molecules. The adeninium cations form N-H...O hydrogen bonds with the phthalate anions. The cations also form infinite one-dimensional polymeric ribbons via N-H...N interactions. In the crystal packing, hydrogen-bonded columns of cations, anions and phthalate anions extend parallel to the c axis. The water molecules crosslink adjacent columns into hydrogen-bonded layers. PMID- 17413230 TI - 1-((E)-{(1R,2R)-2-[(E)-(2-Hydroxy-1 naphthyl)methyleneamino]cyclohexyl}iminiomethyl)naphthalen-2-olate: a Schiff base compound having both OH and NH character. AB - The title Schiff base compound, C(28)H(26)N(2)O(2), possesses both OH and NH tautomeric character in its molecular structure. While the OH side of the compound is described as an intermediate state, its NH side adopts a predominantly zwitterionic form. The molecular structure of the compound is stabilized by both N(+)-H...O(-) and O-H...N intramolecular hydrogen bonds. There are two weak C-H...O hydrogen bonds leading to polymeric chains of topology C(5) and C(13) running along the b axis of the unit cell. In addition, intermolecular C-H...pi interactions serve to stabilize the extended structure. PMID- 17413231 TI - 2,3,5,6-Tetrakis(phenoxymethyl)pyrazine and 2,3,5,6 tetrakis(phenylsulfanylmethyl)pyrazine. AB - The title compounds, C(32)H(28)N(2)O(4), (I), and C(32)H(28)N(2)S(4), (II), respectively, are tetrasubstituted pyrazines and both possess C(i) symmetry. They differ only in the hetero atom (X) of the -CH(2)XPh side-arm substituents: X = O in (I) and S in (II). Compound (I) has an overall S-shape with a pair of adjacent -CH(2)OPh side arms alternately above and below the plane of the central pyrazine ring. The phenyl rings are inclined to one another by 12.63 (18) degrees and there is one intramolecular C-H...O hydrogen bond involving adjacent -CH(2)OPh side arms. In compound (II), adjacent -CH(2)SPh side arms point in opposite directions with respect to the pyrazine ring plane, with the phenyl rings inclined at 60.45 (8) degrees. Both structures have weak C-H...pi intermolecular interactions. PMID- 17413232 TI - 2-Methoxy-4-[(5-oxo-2-phenyl-4,5-dihydro-1,3-oxazol-4-ylidene)methyl]phenyl 4 methylbenzenesulfonate. AB - Molecules of the title compound, C(24)H(19)NO(6)S, adopt the Z configuration and have a distorted tetrahedral geometry around the S atom. The oxazolone, 2-phenyl and methoxyphenyl rings are approximately coplanar. The C atom between the methoxyphenyl and oxazolone rings displays a distorted trigonal bonding geometry. Pairs of molecules are linked into dimers through weak C-H...O hydrogen bonds. PMID- 17413233 TI - 3,4-Diethyl-2,5-dihydro-1H-pyrrole-2,5-dione. AB - In the crystal structure of the title compound, C(8)H(11)NO(2), three distinct molecules are present in the asymmetric unit. The molecules are organized in two different hydrogen-bonded tapes, which form a complex layered structure. A structural comparison with the crystal structures of related maleimide derivatives unravels a stepwise evolution of morphological complexity with increasing molecular complexity for this class of compounds. PMID- 17413234 TI - 4,6-Dimethyl-2-(3-pyridyl)quinolin-5-amine. AB - The title compound, C(16)H(15)N(3), shows a hindrance effect between adjacent amino and methyl groups that leads to a structural distortion, which is reflected in the non-planarity of the quinoline entity and in the bond angles and distances. The crystal packing consists of chains along the b axis sustained by an intermolecular hydrogen bond between the amino group and the N atom of the pyridyl ring. PMID- 17413235 TI - A re-examination of supramolecular aggregation in two polymorphs of acetone 2,4 dinitrophenylhydrazone. AB - The structure of the triclinic polymorph of acetone 2,4-dinitrophenylhydrazone, C(9)H(10)N(4)O(4), has been redetermined from diffraction data collected at 120 (2) K; the molecules are linked by C-H...O hydrogen bonds into centrosymmetric R(2)(2)(10) dimers which are themselves linked into a chain by an aromatic pi-pi stacking interaction. In the monoclinic polymorph, which crystallizes with Z' = 2 in the space group P2(1)/n, one type of molecule forms dimers exactly as in the triclinic polymorph, while the other forms C(6) chains. PMID- 17413236 TI - Ethyl N-[2-(hydroxyacetyl)phenyl]carbamate, ethyl N-[2-(hydroxyacetyl)-4 iodophenyl]carbamate and ethyl N-[2-(hydroxyacetyl)-4-methylphenyl]carbamate. AB - In ethyl N-[2-(hydroxyacetyl)phenyl]carbamate, C(11)H(13)NO(4), all of the non-H atoms lie on a mirror plane in the space group Pnma; the molecules are linked into simple chains by a single C-H...O hydrogen bond. The molecules of ethyl N-[2 (hydroxyacetyl)-4-iodophenyl]carbamate, C(11)H(12)INO(4), are linked into sheets by a combination of O-H...I and C-H...O hydrogen bonds and a dipolar I...O contact. Ethyl N-[2-(hydroxyacetyl)-4-methylphenyl]carbamate, C(12)H(15)NO(4), crystallizes with Z' = 2 in the space group P-1; pairs of molecules are weakly linked by an O-H...O hydrogen bond and these aggregates are linked into chains by two independent aromatic pi-pi stacking interactions. PMID- 17413237 TI - Carbonyl-carbonyl, carbonyl-pi and carbonyl-halogen dipolar interactions as the directing motifs of the supramolecular structure of ethyl 6-chloro-2-oxo-2H chromene-3-carboxylate and ethyl 6-bromo-2-oxo-2H-chromene-3-carboxylate. AB - The title compounds, C(12)H(9)ClO(4), (I), and C(12)H(9)BrO(4), (II), are isomorphous and crystallize in the monoclinic space group P2(1)/c. Both compounds present an anti conformation between the 3-carboxy and the lactone carbonyl groups. Both carbonyl groups are out of the plane defined by the remaining chromene atoms, by 8.37 (6) and 17.57 (6) degrees for (I), and by 9.07 (8) and 18.96 (18) degrees for (II), owing to their involvement in intermolecular interactions. In both compounds, layers of centrosymmetric hydrogen-bonded dimers are developed in the [-5 -2 22] plane through C-H...O interactions, involving both carbonyl groups as acceptors. Two families of dimers stack through C=O...C=O, C=O...pi and C-X...C=O (X = Cl and Br) dipolar interactions, as well as a C-H...pi interaction, developing the three-dimensional structure along the c axis. PMID- 17413238 TI - 2,2,6,6-Tetramethyl-4-oxopiperidinium nitrate. AB - In the title compound, C(9)H(18)NO(+).NO(3)(-), the piperidinium ring adopts a slightly deformed chair conformation and the nitrate anion is disordered. The ions are arranged in hydrogen-bonded chains parallel to [001], in which the cations alternate with the anions. The intrachain hydrogen bonds are bifurcated and link the O atoms of the anions to the N atoms of the cations. PMID- 17413239 TI - 4,4'-Bipyridyl N,N'-dioxide-3-hydroxy-2-naphthoic acid (1/2). AB - 4,4'-Bipyridyl N,N'-dioxide crystallizes with 3-hydroxy-2-naphthoic acid to give a centrosymmetric three-component adduct, C(10)H(8)N(2)O(2).2C(11)H(8)O(3), which is engineered into a two-dimensional layer structure by two kinds of pi-pi interactions. Weak C-H...O interactions further link the two-dimensional structure into a three-dimensional structure. PMID- 17413240 TI - The alpha- and beta-epoxides of 3beta-acetoxy-5alpha-lanost-9(11)-en-7-one. AB - The structures of 3beta-acetoxy-9alpha,11alpha-epoxy-5alpha-lanost-9(11)-en-7-one and 3beta-acetoxy-9beta,11beta-epoxy-5alpha-lanost-9(11)-en-7-one, C(32)H(52)O(4), differ in their respective substituted cyclohexanone rings but adopt similar conformations in the other three rings. In both of the crystal structures, weak intermolecular C-H...O interactions are present. PMID- 17413241 TI - Naphthalene-1,7-diol. AB - The asymmetric unit of the title compound, C(10)H(8)O(2), contains two planar symmetry-independent molecules linked by an O-H...O hydrogen bond. In the crystal structure, molecules are linked into infinite chains of rings, formed by a combination of O-H...O and C-H...O hydrogen bonds, and additionally reinforced by pi-pi stacking interactions. Adjacent chains are connected by weak C-H...pi interactions. PMID- 17413242 TI - Constructor graph description of hydrogen bonding in a supramolecular assembly of N,N'-bis(2-hydroxy-1-methylethyl)phthalamide. AB - Hydrogen bonds of four types (N-H...O=C, N-H...OH, O-H...O=C and O-H...OH) connect molecules of the title compound, C(14)H(20)N(2)O(4), in the crystal into sheets folded into a zigzag pattern. The intermolecular interactions are discussed in terms of the first- through fourth-level graph sets, and a constructor graph helps visualize the supramolecular assembly. PMID- 17413243 TI - 4-Amino-3-methyl-6-phenyl-1,2,4-triazin-5(4H)-one (metamitron) and 4-amino-6 methyl-3-phenyl-1,2,4-triazin-5(4H)-one (isometamitron). AB - The title structures, both C(10)H(10)N(4)O, are substitutional isomers. The N-N bond lengths are longer and the C=N bond lengths are shorter by ca 0.025 A than the respective average values in the C=N-N=C group of asymmetric triazines; the assessed respective bond orders are 1.3 and 1.7. There are N-H...O and N-H...N hydrogen bonds in both structures, with 4-amino-3-methyl-6-phenyl-1,2,4-triazin 5(4H)-one containing a rare bifurcated N-H...N,N hydrogen bond. The structures differ in their molecular stacking and the hydrogen-bonding patterns. PMID- 17413244 TI - Lamellar aggregation and hydrogen-bonding motifs in 3-(aminocarbonyl)pyridinium perchlorate and hydrogen oxalate. AB - In the title compounds, C(6)H(7)N(2)O(+).ClO(4)(-), (I), and C(6)H(7)N(2)O(+).C(2)HO(4)(-), (II), the carboxamide plane is twisted from the plane of the protonated pyridine ring. Lamellar or sheet-like structural features are observed through N-H...O and O-H...O hydrogen-bonded motifs of cations and anions in (I) and (II), respectively. These sheets are aggregated through C(4) and C(5) chain motifs in (I) and (II), respectively. R(1)(2)(4) ring motifs in (I) and R(1)(2)(5) motifs in (II) are formed via pyridine-anion bifurcated N H...O interactions. In (II), carboxamide groups form N-H...O dimers around the inversion centres of the unit cell, with R(2)(2)(8) ring motifs. A 2(1) screw related helical or ribbon-like structure along the b axis is formed in (II) through carboxamide and pyridinium N-H...O hydrogen bonds with the oxalate anions. PMID- 17413245 TI - Differences in the crystal structures of two dialkyl diester triphenylphosphonium ylids. AB - Hydrogen bonding and crystal packing play major roles in determining the conformations of ethyl methyl 2-(triphenylphosphoranylidene)malonate, Ph(3)P=C(CO(2)CH(3))CO(2)CH(2)CH(3) or C(24)H(23)O(4)P, (I), and dimethyl 2 (triphenylphosphoranylidene)malonate, Ph(3)P=C(CO(2)CH(3))(2) or C(23)H(21)O(4)P, (II). In (I), the acyl O atom of the ethyl ester group is anti to the P atom, while the O atom of the methyl ester group is syn. In (II), the dimethyl diester is a 1:1 mixture of anti-anti and syn-anti conformers. PMID- 17413246 TI - [Stents in interventional cardiology]. AB - Since the first percutaneous transluminal coronary angioplasty performed by A. Gruentzig in 1977, percutaneous coronary interventions have become the most important treatment modality for coronary heart disease. Coronary angioplasty carried a significant risk of coronary flow-limiting dissections and restenosis during the first six months following the procedure. Two main studies comparing percutaneous transluminal coronary angioplasty and coronary stenting (STRESS and BENESTENT) performed in 1994 showed a significant reduction in restenosis rate using stents. Thus, until now stents are the most widely used devices for coronary intervention despite two problems: subacute stent thrombosis (1-2%) and still high restenosis rate (5-40%). Subacute stent thrombosis occurs within the first month after stent placement and can be prevented using the double antiplatelet regimen with aspirin and clopidogrel. Some risk of subacute thrombosis remains beyond the first month when drug-eluting stents are used. This requires prolonged antiplatelet therapy. Drug-eluting stents are the most significant innovation in interventional cardiology. They can reduce the incidence of restenosis in native stable coronary arteries to 3-5%. However, the long-term studies comparing bare-metal stents and drug-eluting stents did not show any significant differences in the rate of major adverse cardiac events (death, myocardial infarction), especially in patients with diabetes after the treatment of bifurcational lesions. According to proposed recommendations, drug eluting stents should be used in small vessels, restenotic lesions, and in saphenous vein grafts. Despite some disadvantages, the results of coronary stenting using drug-eluting stents continue to improve. PMID- 17413247 TI - Predicting outcome of treatment with radiotherapy in endocrine ophthalmopathy. AB - OBJECTIVE: To evaluate if the concentration of C-reactive protein and the level of antibodies to thyrotropin receptors might predict the outcome of retrobulbar irradiation in patients with endocrine ophthalmopathy. MATERIAL AND METHODS: Patients with moderate endocrine ophthalmopathy received orbital radiotherapy. The overall severity of endocrine ophthalmopathy was assessed using the total eye score based on the NOSPECS classification before the treatment and 6 months later. Treatment outcome was evaluated using major and minor criteria recommended by L. Bartalena 6 months after the treatment. Patients who improved in at least one major or in two or more minor criteria were considered responders. Patients in whom no changes occurred or who responded in only one minor criterion or eye status worsened were classified as nonresponders. The active disease was considered present in a patient who responded successfully to retrobulbar irradiation, and the inactive one when a patient did not respond. RESULTS: The level of antibodies to thyrotropin receptors in responders was 24.0 IU/L (range 2.0-405.0 IU/L) and in nonresponders 23.0 IU/L (range 2.0-405.0 IU/L); P=0.72. C reactive protein levels in responders and nonresponders were 0.1 mg/L (range 0.1 3.1 mg/L) and 0.1 mg/L (range 0.1-1.5 mg/L), respectively; P=0.92. Although responders and nonresponders differed by proptosis, the severity of endocrine ophthalmopathy, and clinical activity score, but according to the binary logistic regression model, only the clinical activity score could give additional information on the prediction of the treatment outcome. If clinical activity score increased by 1, odds ratio for successful treatment outcome increased 2.4 times. CONCLUSIONS: 1) At the baseline of radiotherapy, the level of antibodies to thyrotropin receptors and concentration of C-reactive protein in responders did not differ from nonresponders; 2) Responders did not differ from nonresponders to radiotherapy by age, gender, duration of endocrine ophthalmopathy and thyroid disease; 3) The pretreatment clinical activity score, total eye score, proptosis of the responders were higher. PMID- 17413248 TI - Effects of the prolonged thoracic epidural analgesia on ventilation function and complication rate after the lung cancer surgery. AB - Thoracic epidural analgesia has been considered to have a good anesthetic efficacy and to decrease the postoperative complication rate, while its effect upon the ventilation function is still the topic of many clinical studies. The aim of this study was to evaluate the course of early postoperative period using thoracic epidural analgesia. MATERIAL AND METHODS: A total of 453 patients undergoing the operation due to the non-small cell carcinoma were selected and examined. Their postoperative complications and mortality rate were evaluated. In 79 patients, arterial oxygen saturation (SaO(2)), forced vital capacity, forced expiratory volume in the first second, and the efficacy of analgesia were analyzed within the first 7 days after the operation. These patients were divided into subgroups according to the type of the operation--lobectomy or pneumonectomy -and the type of analgesia--thoracic epidural analgesia or opiates administered intramuscularly (control group). RESULTS: A better statistically significant efficacy of analgesia was observed in thoracic epidural analgesia group than in the control group (visual analog pain scale score 2.5 versus 5.3, P<0.01). There was also a statistically significant lower incidence of postoperative complications (20.5% versus 38.8%, respectively). Thoracic epidural analgesia is a factor decreasing the relative risk of complications (RR=0.53, 95% CI 0.28 0.99, P=0.0233). In the lobectomy group, 24 hours after the surgery, forced vital capacity was 61+/-12% in the group receiving thoracic epidural analgesia and 45+/ 13% in the control group (P=0.0152); forced expiratory volume in the first second was 56+/-17% and 41+/-11%, respectively (P=0.0308). In the pneumonectomy group, 24 hours after the surgery, forced vital capacity was 47+/-16% in the group receiving thoracic epidural analgesia, 35+/-8% in the control group (P=0.080). Forced expiratory volume in the first second was 47+/-15% and 36+/-7%, respectively (P=0.0449). CONCLUSION: We conclude that analgesia with intramuscularly administered opioids provides unsatisfactory analgesia, especially in the first days after the operation. Thoracic epidural analgesia is a safe method, which provides a better quality of life for the patient, decreases the postoperative complication rate, and improves the ventilation function after the lung operations. PMID- 17413249 TI - Bland-Altman analysis as an alternative approach for statistical evaluation of agreement between two methods for measuring hemodynamics during acute myocardial infarction. AB - BACKGROUND AND OBJECTIVE: Evaluation of hemodynamics in patients with acute myocardial infarction is crucial. In clinical practice, the comparison of a new measurement technique with an established one is often needed to see whether they agree sufficiently for the new to replace the old. Such investigations are often analyzed inappropriately, notably by using correlation coefficients. Our objective is to present an alternative approach, suggested by D. G. Altman and J. M. Bland, based on graphical techniques and simple calculations, for evaluation of the agreement of two methods--intermittent thermodilution (recognized and accepted as a "gold standard" for hemodynamic monitoring) and impedance cardiography (newly introduced method). PATIENTS AND METHODS: A total of 34 patients (20 (58.8%) men and 14 (41.2%) women) were investigated according to the study protocol at Kaunas University of Medicine Hospital. A prospective controlled study was designed to compare two different methods--intermittent thermodilution and impedance cardiography--of cardiac output measurement simultaneously in patients with acute myocardial infarction. Statistical analysis was performed with Bland-Altman and linear regression. RESULTS: A total of 34 paired measurements were carried out in 34 patients at the initiation of hemodynamic monitoring and 30 paired measurements in 32 patients after subsequent improvement or stabilization of clinical status. Correlation coefficient ranged from 0.37 to 0.98. CONCLUSIONS: Bland-Altman analysis is an alternative method for assessing the agreement between two methods of clinical measurement. According to our data, noninvasive technique--impedance cardiography--is a reliable method for hemodynamic monitoring in noncomplicated cases of acute myocardial infarction. PMID- 17413250 TI - Exeter total hip arthroplasty with matte or polished stems. AB - OBJECTIVE: To compare implant survival rates after total hip arthroplasty with Exeter matte or polished stems and to determine the relationship of synovitis/joint effusion to signs of implant loosening and stem type. MATERIAL AND METHODS: The first part of the study included retrospective revision rate analysis of 118 primary hip replacements performed during 1991-1995. Two different designs of Exeter stems were used: matte surface stems during 1991-1995 (matte surface group--47 cases), and polished stems during 1992-1995 (polished stem group--71 cases). During the second part of the study, 24 patients (11 in polished stem group and 13 in matte stem group) were prospectively examined with radiography and sonography. Sonography was performed in order to evaluate capsular distension, i.e. the distance between prosthetic femoral neck and anterior capsule. Capsular distension depends on synovitis and/or synovia in prosthetic hip. The relationship between capsular distension, stem type, and radiographic signs of loosening was assessed. RESULTS: For the first part of our study, total implant survival was 78% with matte stems and 61% with polished stems 13 years postoperatively (P=0.27). Stem survival was 82% for matte stems, and 88% for polished stems (P=0.54). In the second part of study, a significant relationship between increased capsular distension and cup loosening was determined (P=0.04). We did not find significant difference in capsular distension when compared matte and polished stems. CONCLUSION: Implant survival rates did not differ between the groups. The relationship between capsular distension and cup loosening was statistically significant. PMID- 17413251 TI - [The diuretic effect of cornflower water extract]. AB - The objective of this study was to evaluate and compare the effect of cornflower water extract and hydrochlorothiazide on diuresis, Na(+) and K(+) excretion, and the changes in the prostaglandin E(2) and kinins levels in the blood plasma. MATERIAL AND METHODS: Male Wistar rats were used in all experiments. Animals were divided into nine groups. Diuretic activity was investigated according to the technique proposed by E. B. Berchin; urinary electrolyte contents were analyzed by flame photometry. Prostaglandin E(2) content was measured by radioimmunoassay using (3)H isotopes, kinins--by enzymatic methods. RESULTS: It has been established that in male rats, receiving hydrochlorothiazide, the volume of urine excreted two and four hours after the administration of the drug was by 18% and 17%, respectively, higher as compared to the rats that were given cornflower water extract (P<0.05). The diuretic effect of cornflower water extract was noted in the animal group receiving this extract as compared to the control group: after two hours, the volume of urine excreted increased from 2.03+/-0.03 mL to 2.44+/-0.04 mL and after four hours--from 3.88+/-0.07 mL to 5.35+/-0.1 mL. Administration of hydrochlorothiazide under the load of salts and water resulted in a higher excretion of sodium and potassium as compared to the effect of cornflower water extract. The highest prostaglandin levels were found in the blood plasma of the animals receiving hydrochlorothiazide. Under the load of salts and water, a 13% and 15% increase, respectively, in the amount of prostaglandins was observed in the animals that were given cornflower water extract compared to the control animals (P<0.05). The greatest increase in the amount of kinins was found in the groups of animals that were given hydrochlorothiazide under the load of salts and water (14% and 22%, respectively). Kinin levels did not differ statistically significantly between the control group and the groups receiving cornflower water extract. CONCLUSION: Cornflower water extract possesses diuretic activity, but its effect was lower than that of hydrochlorothiazide. PMID- 17413252 TI - Bimodal recovery of quadriceps muscle force within 24 hours after sprint cycling for 30 seconds. AB - The aim of the study was to investigate the manifestation of potentiation and fatigue as well as the coexistence of these phenomena at different muscle lengths during a 24-hour period after a sprint cycling for 30 s. MATERIAL AND METHODS: Twelve healthy untrained men (mean age 23.6+/-1.7 years) took part in the experiment. The contractility of quadriceps muscle was studied before (Initial) and 2, 5, 30, 60 min and 24 h after exercise via the electrically evoked contractions at 1, 15, 50 Hz and maximal voluntary contractions at short and long muscle length. RESULTS: 1) In early, fast-recovery phase (within the first 5 min), muscle force evoked by electrical stimulation of 1, 15, 50 Hz was restored at short muscle length, conversely at long length (Initial vs. 5 min: 15 Hz and 50 Hz, both P<0.05), whereas maximal voluntary contraction force was still suppressed at both muscle lengths; 2) in the second phase (from 5 min to 30-60 min), muscle force decreased at low- and high-frequency stimulations and was more expressed at low-frequency stimulation and at short muscle length than that at long length, but the maximum voluntary contraction force recovered to initial; 3) in long-lasting phase (within 24 hours), 15 Hz force was still suppressed at both muscle lengths. CONCLUSION: A bimodal recovery of contractility of the quadriceps following sprint cycling for 30 s is determined by the concomitant complex interaction of mechanisms enhancing (potentiation) and suppressing (fatigue) contractile potential of the muscle. PMID- 17413253 TI - Outpatient methadone maintenance treatment program. Quality of life and health of opioid-dependent persons in Lithuania. AB - BACKGROUND: The evaluation of quality of life and self-perceived health represents an assessment of the impact of treatment on patient functioning and well-being. OBJECTIVE: Our aim was to explore the impact of methadone maintenance treatment on quality of life and self-perceived health of opioid-dependent persons in Lithuania. METHODS: A total of 102 opioid-dependent patients were recruited in the study. A prospective follow-up study design was used. To assess quality of life, the WHOQOL-BREF 26-item version was used. The impact of methadone maintenance treatment on self-perceived health was assessed by Opiate Treatment Index (OTI). RESULTS: Following 6 months of methadone maintenance treatment, significant improvements in physical (P=0.004), psychological (P=0.004), and environmental (P=0.048) components of quality of life were observed; no statistically significant improvements were found in social component of quality of life. Study participants reported lower rates of medical morbidity associated with injection (P<0.001), cardiorespiratory (P=0.034), musculoskeletal (P<0.001), neurological (P=0.013), gastrointestinal (P<0.001), and general health (P<0.001). CONCLUSIONS: Methadone maintenance treatment substantially reduces morbidity associated with opioid dependence and improves the quality of life of patients. PMID- 17413254 TI - The prevalence of type 1 diabetes mellitus among adolescents and adults in Lithuania during 1991-2004. AB - OBJECTIVE: The goal of this study was to summarize the data on the prevalence of type 1 diabetes mellitus among Lithuanian population aged more than 15 years. MATERIAL AND METHODS: The data on patients aged more than 16 years were collected with the help of general practitioners, endocrinologists, and physicians internists working in the diabetes care in all towns and regions of Lithuania. The data on patients aged 14 to 16 years were obtained from the National Register of Diabetes Mellitus in Childhood in Lithuania. RESULTS: In Lithuania, on December 31, 1991, there were 2179 adolescent and adult patients with type 1 diabetes mellitus or 75.21 per 100,000 inhabitants of the same age group (95% Poisson CI 72.12-78.43), and at the end of 2004--3996 or 140.69 (95% Poisson CI 136.40-145.12), respectively. During a 14-year period, the mean increase in the number of type 1 diabetic patients was 144.85+/-23.32 persons per year or 4.66+/ 1.17% or 4.04+/-1.19 cases per 100,000 population (for males 85.54+/-10.82 or 5.06+/-1.02% or 6.81+/-1.57/100 000 and for females 54.23+/-9.05 or 3.93+/-0.86% or 3.56+/-1.05/100,000). Regression-based linear trends showed that the prevalence of type 1 diabetes mellitus among population aged more than 15 years had a tendency to increase. The prevalence rates of type 1 diabetes mellitus among adolescent and adult subjects, adjusted for Lithuanian male and female age groups, were 80.64/100,000 and 70.23/100,000 in 1991 (P<0.05) and 166.52 and 117.63 in 2004 (P<0.05), respectively. CONCLUSIONS: The prevalence of type 1 diabetes mellitus among Lithuanian females aged more than 15 years was lower than among males. The register provides the possibility of highly precise collection of the data on patients from various medical care units in Lithuania. PMID- 17413255 TI - [Changes in morphometric parameters and function of left ventricle in child and adolescent athletes]. AB - The aim of the study was to assess the impact of physical load on left ventricular morphometric parameters and function in child and adolescent athletes. MATERIAL AND METHODS: A total of 143 trained athletes aged 7-17 years and 54 healthy nonathletic children and adolescents aged 8-17 years were involved in this study. The participants were divided into four groups according to the duration of physical activity (training hours per week). Two-dimensional, M-mode, and Doppler echocardiography were used to evaluate cardiac dimensions and function. Absolute parameters and parameters corrected for body surface area were calculated. Left ventricular fractional shortening was calculated as an index of systolic function, and E/A ratio was calculated for evaluation of left ventricular diastolic function. RESULTS: In 69.9% of athletes, septal and posterior wall thickness, end-diastolic diameter, left ventricular mass, and mass index were statistically significantly higher than in controls. There were no differences in left ventricular end-diastolic diameter and posterior wall thickness corrected for body surface area as well as diastolic E/A ratio between the groups. The fractional shortening in athletes was significantly higher (P<0.01). Interventricular septum thickness, end-diastolic diameter, and left ventricular mass were significantly higher in athletes whose training exceeded 8 hours per week compared to the controls. Left ventricular fractional shortening was significantly higher in athletes training more than 10 hours per week than in controls. Diastolic function index--E/A ratio--did not differ between the groups. CONCLUSION: Our study demonstrated that echocardiographic parameters of child and adolescent athletes statistically significantly exceeded the parameters of untrained controls. These parameters were dependent on the anthropometric data and physical activity (the duration of training expressed in hours per week). PMID- 17413256 TI - [Electrical injuries]. AB - Electrical trauma can be caused by low-voltage current (from 60 to 1000 V, usually 220 or 360 V), high-voltage (more than 1000 V) current, lightning, and voltaic arc. Often victims are little children, teenagers, and working-age adults. Electrical injuries and clinical manifestations can vary a lot and range from mild complaints not demanding serious medical help to life-threatening conditions. Lightning causes serious injuries in 1000-1500 individuals every year worldwide. The case fatality rate is about 20-30%, with as many as 74% of survivors experiencing permanent injury and sequela. The primary cause of death in victims of lightning strike or other electrical trauma is cardiac or respiratory arrest. That is why appropriate urgent help is essential. Subsequently electrical burns, deep-tissue and organ damage caused by electricity, secondary systemic disorders often demand intensive care and prompt, usually later multistage surgical treatment; therefore, prevention of electrical trauma, which would help to reduce electrical injuries in children and working age population, is very actual. The most important is to understand the possible danger of electricity and to avoid it. PMID- 17413257 TI - Design and concepts of the NIMH Collaborative HIV/STD Prevention Trial. PMID- 17413260 TI - Selection of populations represented in the NIMH Collaborative HIV/STD Prevention Trial. AB - OBJECTIVE: To identify venues with vulnerable populations suitable for testing the community popular opinion leader intervention in each of the five countries (China, India, Peru, Russia, and Zimbabwe) participating in the National Institute of Mental Health (NIMH) Collaborative HIV/STD Prevention Trial. DESIGN: HIV epidemiology and vulnerable populations differ considerably across the countries. Therefore, different community populations were targeted in the five countries. METHODS: Venues and populations were chosen on the basis of specific selection criteria (investigated during the Trial's ethnographic research phase): the willingness of stakeholders and gatekeepers of the venues to cooperate; geographical boundaries defining each venue; population stability within venues; the independence of venues and non-overlap of population members across multiple venues; population size within each venue; social interaction opportunities; and either a high level of sexual risk behavior or a high prevalence of sexually transmitted diseases (STDs) or HIV. RESULTS: Venues and populations selected were food market stall owners and workers in China, male patrons of wine shops and at risk women congregating near the shops in India, young men and women in social gathering points in neighborhoods in Peru, trade and vocational school dormitory residents in Russia, and people congregating in growth points in Zimbabwe. CONCLUSION: Although the target populations differed across countries, they shared in common high behavioral or biological risk at baseline and suitability for a randomized trial of a community-level HIV/STD prevention behavioral intervention. PMID- 17413261 TI - Challenges and processes of selecting outcome measures for the NIMH Collaborative HIV/STD Prevention Trial. AB - OBJECTIVE: To review the challenges of designing behavioral and biological outcome measures for the multinational NIMH Collaborative HIV/STD Prevention Trial and provide the rationale for selecting these measures. DESIGN: Although many different evidence-based prevention programmes have been developed, few have been evaluated in different countries, cultures, and populations. One issue in evaluating the generalized efficacy of any prevention approach is to identify a set of common outcome measures useful across diverse settings and peoples. The Trial is designed to evaluate whether the community popular opinion leader intervention can be adapted cross-nationally and cross-culturally for different populations and still retain its efficacy. METHODS: Literature reviews, investigator experience, ethnographic study, pilot studies, and epidemiological studies were used to select the endpoints for the Trial. RESULTS AND CONCLUSION: Both biological and behavioral data will be obtained at baseline and 12 and 24 months post-baseline. Communities that receive the intervention will be compared with matched control communities on two primary outcomes: (i) a change in self reported unprotected sexual acts with non-spousal, non-live-in partners; and (ii) the incidence of sexually transmitted disease (STD), defined as a composite index of viral and bacterial STD. PMID- 17413262 TI - Methodological overview of a five-country community-level HIV/sexually transmitted disease prevention trial. AB - OBJECTIVE: To provide an overview of the National Institute of Mental Health (NIMH) Collaborative HIV/STD Prevention Trial taking place in five populations at risk of HIV and sexually transmitted diseases in China, India, Peru, Russia, and Zimbabwe, including the rationale, study management, methods, and proposed data analyses. DESIGN: The Trial will scientifically evaluate the effectiveness of the community popular opinion leader (C-POL) community-level HIV prevention intervention that was adapted for use in the various cultures within the resource limitations faced by service providers in world regions threatened by high rates of HIV infection. METHODS: The study phases consist of an ethnographic study, pilot studies, an epidemiological study, and a community-randomized trial. The Trial uses the C-POL intervention, which researchers selected on the basis of research that shows the intervention's success in populations vulnerable to HIV risk behavior in the United States, and has the potential to be applied in a variety of international settings. RESULTS: Trial results will be tabulated by and across country by randomization assignment. Results will include a careful review of data to substantiate original assumptions used in the study design. Data collection will not conclude until August 2007. CONCLUSION: Although data collection is incomplete, researchers have learned lessons throughout the development of the study. These include the importance of preliminary epidemiological studies; the close monitoring of biological testing, follow-up rates and process measures at international sites; the tailoring of assessments and interventions to various cultures; regular communication; and a review of the timeline to accommodate Institutional Review Board clearances. PMID- 17413263 TI - Design and integration of ethnography within an international behavior change HIV/sexually transmitted disease prevention trial. AB - OBJECTIVE: To use a common ethnographic study protocol across five countries to provide data to confirm social and risk settings and risk behaviors, develop the assessment instruments, tailor the intervention, design a process evaluation of the intervention, and design an understandable informed consent process. DESIGN: Methods determined best for capturing the core data elements were selected. Standards for data collection methods were established to enable comparable implementation of the ethnographic study across the five countries. METHODS: The methods selected were participant observation, focus groups, open-ended interviews, and social mapping. Standards included adhering to core data elements, number of participants, mode of data collection, type of data collection instrument, number of data collectors at each type of activity, duration of each type of activity, and type of informed consent administered. Sites had discretion in selecting which methods to use to obtain specific data. RESULTS: The ethnographic studies provided input to the Trial's methods for data collection, described social groups in the target communities, depicted sexual practices, and determined core opinion leader characteristics; thus providing information that drove the adaptation of the intervention and facilitated the selection of venues, behavioral outcomes, and community popular opinion leaders (C-POLs). CONCLUSION: The described rapid ethnographic approach worked well across the five countries, where findings allowed local adaptation of the intervention. When introducing the C-POL intervention in new areas, local non governmental and governmental community and health workers can use this rapid ethnographic approach to identify the communities, social groups, messages, and C POLs best suited for local implementation. PMID- 17413264 TI - The feasibility of audio computer-assisted self-interviewing in international settings. AB - OBJECTIVE: To determine the feasibility of using audio computer-assisted self interviewing (ACASI) for data collection in developing countries, and to compare responses to questions eliciting sensitive information about sexual behavior using ACASI versus computer-assisted personal interviewing (CAPI) in five developing countries. DESIGN: A feasibility study determined whether ACASI could be used in populations in developing countries. A follow-up, randomized crossover study compared responses to questions eliciting sensitive information about sexual behavior using ACASI versus CAPI. METHODS: The NIMH Collaborative HIV/STD Prevention Trial conducted a feasibility study of ACASI in convenience samples in China, India, Peru, and Russia, then a randomized crossover ACASI versus CAPI study among volunteers in these countries plus Zimbabwe. RESULTS: Approximately equal numbers of men and women completed the feasibility study; the results suggested a high comfort level among participants. Married respondents in China and India appeared to give unreliable responses on sexual activity. In the crossover study, the pattern of responses to sensitive questions showed few differences. In China, higher rates of sexual risk were reported on CAPI. In Peru and Russia, differences by mode were found in the number of partners in the past year. CONCLUSION: Despite variable computer experience and literacy, feasibility study participants reported ease in completing ACASI, and preferred a computer to an interviewer for answering sensitive questions, or had no preference. In the crossover study, most participants gave similar responses on both modes of survey administration. ACASI appears to be feasible in these settings, although low literacy may pose problems if participants cannot clarify questions. PMID- 17413265 TI - The community popular opinion leader HIV prevention programme: conceptual basis and intervention procedures. AB - OBJECTIVE: To describe the community popular opinion leader (C-POL) intervention employed in the NIMH Collaborative HIV/STD Prevention Trial, including its theoretical, conceptual, and empirical basis, intervention procedures and methods, core elements, and how its content was culturally tailored to address the needs of varied populations. DESIGN: The programme is designed to identify, recruit, train, and intensively engage C-POLs of a target population to convey HIV risk reduction messages to people in their communities, with the intention of reducing high-risk behavior at a population level. METHODS: Based on the diffusion of innovation theory, the intervention identified, trained, and engaged C-POL within a high-risk community population to advocate, recommend, and endorse the importance of safer behavior to other members of the same population. Nine core elements of the intervention are discussed. Data collected during rapid ethnography were used to adapt the content of the intervention for food market owners and workers in China, male patrons of wine shops and at-risk women congregating nearby in India, young people in social gathering venues in Peruvian barrios, dormitory students in Russia, and people congregating in commercial areas of growth points in Zimbabwe. RESULTS: The C-POL intervention model taps into community strengths, altruism, and people's desire to do something to help fight against AIDS. With few exceptions, C-POLs participated enthusiastically in the training sessions and reported having conversations in the community. CONCLUSION: Rapid ethnography can be used to tailor an intervention to diverse settings while maintaining fidelity to the core elements of the intervention. PMID- 17413266 TI - Ethical issues in the NIMH Collaborative HIV/STD Prevention Trial. AB - OBJECTIVE: To develop decision rules regarding key ethical dimensions in scientific protocols for the National Institute for Mental Health (NIMH) Collaborative HIV/STD Prevention Trial taking place in five countries (China, India, Peru, Russia, and Zimbabwe). DESIGN: Countries had HIV rates from 27 to 0.1%, the standard of care varied from access to antiretroviral drugs to no availability, and the reporting of sexually transmitted diseases (STD) to government agencies was mandatory in some countries and not in others. These variations presented challenges when developing decision rules that could be uniformly adopted across countries and simultaneously follow the ethical principles of beneficence, respect, and justice. METHODS: We used several strategies to identify and resolve ethical dilemmas for this international HIV prevention trial. First, we identified key principles, especially those derived for clinical therapeutic, biomedical preventive, or device trials. We convened a 'workgroup on protecting human participants' and charged them with identifying and implementing optimal procedures for ensuring the ethical and equitable treatment of participants and making recommendations to minimize physical, psychological, and social harm to the participants. Each site had a community advisory board, essential in identifying local ethical issues and possible resolutions to them. The NIMH established a data safety and monitoring board with ultimate responsibility for adjudicating ethical dilemmas and decisions. The protocols were deliberated thoroughly by the Trial steering committee, and approved by nine United States and five in-country institutional review boards. RESULTS: We summarize the decision rules adopted to resolve the ethical dilemmas identified. Especially important were the translation of clinical trials principles for a behavioral intervention trial, strategies for ensuring confidentiality and informed consent, dilemmas relating to partner notification of sexually transmitted infections including HIV, minimizing the risks of social harm, establishing community partnerships, ensuring equity among United States and in-country principal investigators, and building capacity for additional research. CONCLUSION: We document our processes and decisions, and their underlying rationales, and hope they contribute to the development of further thinking and practice regarding the ethics of social and behavioral HIV and STD prevention trials in resource-poor settings. PMID- 17413267 TI - Sexually transmitted disease and HIV prevalence and risk factors in concentrated and generalized HIV epidemic settings. AB - BACKGROUND: In many developing countries, the threat of nascent HIV epidemics expanding rapidly requires immediate and appropriate HIV prevention activities. Inexpensive and sustainable interventions are especially relevant in resource constrained environments. In 2001, we assessed the prevalence and behavioral risk of sexually transmitted disease (STD) and HIV among at-risk populations in five developing countries in preparation for a community-randomized controlled trial, the NIMH Collaborative HIV/STD Prevention Trial. METHODS: Using a standardized protocol, more than 1000 participants in each country (China, India, Peru, Russia, and Zimbabwe) were selected by random sampling methods, completed a behavioral risk assessment, and provided biological specimens using a common laboratory protocol. Sample characteristics were studied within each country, and risk factors for HIV/STD acquisition were evaluated using logistic regression models. RESULTS: HIV rates were low (<1%) in China, India, Peru, and Russia but were high (26%) in rural Zimbabwe. STDs were generally twice as common in women as men, and serological evidence of herpes simplex virus type 2 infection was the most frequently detected STD. Behavioral data showed high rates of multiple partners in the Russian sample, and very low condom use rates in India and China. Among participants who reported ever having sex, female sex and having two or more sex partners were the factors most frequently associated with an increased risk of prevalent STD. CONCLUSION: Behavioral or biological risks were of sufficient magnitude in the locations selected in China, Russia, and Zimbabwe to implement the community-based randomized trial. Higher-risk subsets of community residents in India and Peru were identified before beginning the Trial. PMID- 17413268 TI - Formative study conducted in five countries to adapt the community popular opinion leader intervention. AB - OBJECTIVE: To obtain information about the social and cultural factors related to health behaviors influencing HIV/sexually transmitted disease (STD) transmission in study communities in China, India, Peru, Russia, and Zimbabwe so that the assessment and intervention of the National Institute for Mental Health (NIMH) Collaborative HIV/STD Prevention Trial could be adapted appropriately. METHODS: Field observations, focus groups, in-depth interviews with key informants, and an observation of community social dynamics were conducted as part of a rapid ethnographic assessment. RESULTS: All five sites reported a power dynamic tilted towards men, which rendered women particularly vulnerable to HIV and other STDs. Women's relative lack of power was exemplified by a double standard for extramarital sex, women's limited ability to negotiate sex or condom use, and sexual and physical violence against women. In all sites except Russia, extramarital sex is tolerated for men but proscribed for women. In Peru, power dynamics between men who have sex with men were tilted towards men who self identified as heterosexual. Condom use (reported to be low across all sites) was often linked to having sex with only those perceived as high-risk partners. Regardless of site or study population, participants agreed on the following characteristics of an ideal community popular opinion leader (C-POL): respectable, credible, experienced (life and sexual), trustworthy, empathetic, well-spoken, and self-confident. CONCLUSION: The ethnographic studies provided critical information that enabled the study teams to adapt elements of the Trial in culturally appropriate ways in diverse international settings. PMID- 17413269 TI - Role of the data safety and monitoring board in an international trial. AB - OBJECTIVE: To describe the composition and role of the data safety and monitoring board (DSMB) for the National Institute of Mental Health (NIMH) Collaborative HIV/STD Prevention Trial. DESIGN: NIMH appointed to the DSMB nine members representing the following areas of expertise: prevention science, ethnography, infectious diseases (especially HIV and sexually transmitted diseases), laboratory diagnostics, clinical practice, methodology, international trial experience, statistics, and ethics. METHODS: The DSMB assessed the overall study for any concern about plans or implementation and reviewed cumulative study data to evaluate the safety of study participants, the ongoing conduct of the study, and the scientific validity and integrity of the Trial. Because of the Trial's international scope, the DSMB examined the effects of cultural differences on study implementation and fidelity. RESULTS: Among the DSMB recommendations that strengthened the Trial was one to conduct initial epidemiological studies of the venues selected for the intervention to verify risk and to establish intraclass correlation coefficients that could be used to calculate appropriate sample sizes. CONCLUSIONS: The DSMB played a critical role in this Trial. Because members have the expertise required to monitor the Trial, are not involved in the daily management of the Trial, and can review interim analyses and adverse event reports, they are in an excellent position to provide expert advice to ensure that the Trial's goals are achieved and that NIH funds are well invested. PMID- 17413271 TI - The Brugada syndrome. AB - PURPOSE OF REVIEW: The Brugada syndrome has been an area of intensive investigation since its earliest description in 1992, both on a clinical and on a basic research level. In this review, we will focus on recent achievements in the molecular dissection of the disease pathophysiology and on large multicenter studies dealing with prognostic markers and the natural history of the Brugada syndrome. RECENT FINDINGS: In the past year, two additional genetic pathways have been associated with the disease. Also, an inflammatory or infectious etiology has recently been linked with the Brugada syndrome. The debate on the predictive role of programmed electrical stimulation is still ongoing. Very recently, large follow-up studies questioned the prognostic role of programmed electrical stimulation in this disease. SUMMARY: Knowledge on the genetic determinants of the Brugada syndrome remains limited. Therefore, the management and the risk stratification of patients should be performed on a clinical basis. Sufficient evidence exists to reassure clinicians who feel reluctant to include programmed electrical stimulation in the risk stratification strategy of asymptomatic Brugada syndrome patients. PMID- 17413272 TI - Molecular genetics of mitral valve prolapse. AB - PURPOSE OF REVIEW: Mitral valve prolapse is a common disorder with a strong hereditary component. It is associated with important mitral regurgitation requiring surgical repair and other clinical complications. Genetic studies can provide clues to mechanism and therapy. RECENT FINDINGS: Advances in phenotypic classification have led to linkage to sites on chromosomes 11, 13 and 16 and identification of the first mutation in familial mitral valve prolapse not related to connective tissue syndromes - an X-linked filamin A mutation. New understanding of mechanism based on studies in a mouse Marfan model emphasize the dynamic interplay of differentiating cells and growth factors, with strong potential for therapy. SUMMARY: This new knowledge brings us closer to the ultimate goal of preventing the progression of mitral valve disease to the stage of clinical expression. PMID- 17413273 TI - A review of the genetics of essential hypertension. AB - PURPOSE OF REVIEW: Essential hypertension affects more than 20% of the adult population, and has a multifactorial origin arising from an interaction between susceptibility genes and environmental factors. Several strategies have been used to identify hypertension susceptibility genes. This review highlights recent efforts in genetic dissection of essential hypertension. RECENT FINDINGS: Recently, further chromosomal regions harboring blood pressure loci have emerged in genome-wide linkage studies. Findings from a new systematic two-dimensional genome scan are presented, as well as sex-specific loci linked to hypertension in inbred rodent models. Many case-control association studies have been carried out, but results so far have been equivocal. This review discusses some interesting studies combining linkage and association strategies using gene-gene interactions, and studies the use of haplotypes instead of SNPs. Two novel hypertension susceptibility genes are presented, and a short summary on new insights into genes of the renin-angiotensin and adrenergic systems is given. SUMMARY: To date, linkage and association studies have not been convincing. Genome-wide association studies may prove to be an effective approach to the problems posed by complex traits. Combined with candidate gene approaches, it is hoped this strategy will yield convincing evidence for genes associated with essential hypertension. PMID- 17413274 TI - Molecular genetics of arrhythmogenic right ventricular cardiomyopathy: emerging horizon? AB - PURPOSE OF REVIEW: Recent developments in the elucidation of genes underlying arrhythmogenic right ventricular cardiomyopathy and possible pathogenic mechanisms will be highlighted. RECENT FINDINGS: The cardiac desmosome is a multiprotein structure involved in cell-cell interactions. Mutations in genes encoding desmosomal proteins such as PKP2, DSP, JUP, DSC2 and DSG2 underlie arrhythmogenic right ventricular cardiomyopathy, which can therefore be considered a desmosome cardiomyopathy. Mutations in the plakophilin-2 gene are most prevalent. Current pathophysiological insights suggest a final common pathway in which plakoglobin release from the desmosome, independent of the primarily affected desmosomal protein, results in desmosome impairment, intercalated disc remodeling and Wnt/beta-catenin pathway signaling defects. The recognition of left ventricular involvement associated with mutations in desmosomal protein genes and low penetrance suggests that formal criteria should not be followed too closely in selecting patients for DNA analysis, because finding a mutation may have important implications for clinical practice. SUMMARY: Recent developments have demonstrated that arrhythmogenic right ventricular cardiomyopathy can be considered a desmosome cardiomyopathy. Left ventricular involvement is not uncommon in this type of cardiomyopathy. Such findings are important for diagnostics and family screening and form a starting point for the elucidation of other (non)-genetic factors influencing disease progression and outcome. PMID- 17413275 TI - Genetics of hypertrophic cardiomyopathy: one, two, or more diseases? AB - PURPOSE OF REVIEW: Hypertrophic cardiomyopathy is the most common identifiable cause of sudden death in the young. This review details the history of hypertrophic cardiomyopathy, recent discoveries in its genetic underpinnings and important genotype-phenotype relationships described in recent studies. RECENT FINDINGS: Since the discovery of the genetic underpinnings of hypertrophic cardiomyopathy in 1989 hundreds of mutations scattered among at least 10 sarcomeric genes confer the pathogenetic substrate for this 'disease of the sarcomere/myofilament'. More recently, the genetic spectrum of hypertrophic cardiomyopathy has expanded to encompass mutations in Z-disc-associated genes (Z disc hypertrophic cardiomyopathy) and glycogen storage diseases mimicking hypertrophic cardiomyopathy (metabolic hypertrophic cardiomyopathy). Recent genotype-phenotype studies have discovered an important relationship between the morphology of the left ventricle, its underlying genetic substrate and the long term outcome of this disease. SUMMARY: Genomic medicine has entered clinical practice and the diagnostic utility of genetic testing for hypertrophic cardiomyopathy is clearly evident, but with the growing number of hypertrophic cardiomyopathy-associated genes strategic choices have to be made. With recent discoveries in genotype-phenotype relationships, especially pertaining to the echocardiographic septal shape and the underlying pathogenetic mutation, time has come to subdivide the one disease we call hypertrophic cardiomyopathy. PMID- 17413276 TI - The genetics of congenital heart disease: a review of recent developments. AB - PURPOSE OF REVIEW: As our understanding of the molecular regulation of cardiac development has progressed, an increasing number of genes that cause congenital heart disease when mutated are being identified. This review focuses on the progress made during the past year. RECENT FINDINGS: After PTPN11 was identified as a Noonan syndrome disease gene, additional discoveries have made clear that mutations in other genes along the RAS signaling pathway can cause a spectrum of syndromes and possibly isolated congenital heart disease. Similarly, alterations of genes in other signaling and transcriptional pathways may contribute to the development of atrial septal defects and bicuspid aortic valves. Recently identified disease genes for syndromes associated with congenital heart disease are also reviewed. Finally, the possibility that somatic mosaicism may contribute to the development of congenital heart disease is discussed. SUMMARY: The recent knowledge about the molecular genetic causes of congenital heart disease is reviewed. In many instances, these gene discoveries are being rapidly translated into meaningful genetic testing, which is improving the diagnosis and prognostication for congenital heart disease in isolation or in the context of a syndrome. Ultimately, genetic information will be necessary for planning care as well as clinical research. PMID- 17413277 TI - Clinical approaches to the diagnosis of acute heart failure. AB - PURPOSE OF REVIEW: Predicting which patients with congestive heart failure will decompensate is often difficult, and it is often difficult to distinguish congestive heart failure from other causes of acute dyspnea. This review will focus on some of the newer tools used to diagnose acute congestive heart failure in addition to reviewing the utility of more traditional tools. RECENT FINDINGS: The integration of pertinent positives and negatives on a routine history, key physical findings on examination and routine noninvasive imaging offers high positive and negative predictive power for the diagnosis of acute heart failure. Measurement of B-type natriuretic peptide and N-terminal proB-type natriuretic peptide offers additional and incremental diagnostic information. Measurement of intrathoracic impedance is a novel and potentially useful tool to track absolute changes in cardiac function and total lung fluid content, and may be useful for the outpatient titration of medical therapy to minimize acute congestive heart failure decompensation. SUMMARY: Consistent accurate diagnosis of decompensated congestive heart failure is possible using no more than a complete history and physical examination along with routine imaging techniques. The ability to diagnose acute congestive heart failure however, is improved by using serum B type natriuretic peptide and intrathoracic impedance, both of which offer additive and complementary diagnostic information. PMID- 17413278 TI - The contribution of familial and heritable risks in heart failure. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the recent literature regarding the familial heritability of heart failure and to discuss the possible mechanisms through which this risk is mediated. RECENT FINDINGS: Data from the Framingham Heart Study recently showed that the parental occurrence of heart failure increases the risk of heart failure in offspring. Although the mechanisms mediating this increased risk are not elucidated, heritable risks of heart failure may result from genes affecting the cardiac or vascular systems. Alternatively, familial risk may be mediated partly through the inheritance of recognized or as yet unidentified risk factors for heart failure. Heritable components or genetic loci for quantitative traits contribute to the development of hypertension, coronary artery disease, cardiomyopathies, valvular heart disease, and metabolic conditions, which collectively increase the risk of heart failure. SUMMARY: A careful assessment of the family history of heart failure and associated risk factors may identify treatable targets that can potentially reduce the likelihood of developing heart failure, and can assist in the implementation of preventive strategies for risk populations with stages A and B heart failure. PMID- 17413279 TI - Use of pulmonary artery catheters in advanced heart failure. AB - PURPOSE OF REVIEW: Randomized trials have demonstrated that use of the pulmonary artery catheter does not improve outcomes in patients with critical illness. Despite this, pulmonary artery catheters continue to be used, especially in heart failure. This review addresses recent clinical trials evaluating the safety and efficacy of the pulmonary artery catheter. It also offers guidelines on when pulmonary artery catheter use could be considered in patients with advanced heart failure. RECENT FINDINGS: A meta-analysis demonstrated that use of the pulmonary artery catheter in critically ill patients had no significant impact on morbidity or mortality. In the Evaluation Study of Congestive Heart Failure and Pulmonary Artery Catheterization Effectiveness trial, the addition of the pulmonary artery catheter to careful clinical assessment also had neutral impact. The results of the study are limited, however, by the patient population that was selected for the study, and there still may be a role for the pulmonary artery catheter in advanced heart failure. SUMMARY: Based on the results of the Evaluation Study of Congestive Heart Failure and Pulmonary Artery Catheterization Effectiveness trial and the meta-analysis, there is no indication for routine use of pulmonary artery catheters to adjust therapy during hospitalization for decompensation of chronic heart failure. There remain many specific clinical situations in heart failure in which the pulmonary artery catheter may be useful, however. PMID- 17413280 TI - Bibliography. Current world literature. Molecular genetics. PMID- 17413282 TI - Nutrition support in benign and malignant disease: a practical guide. PMID- 17413283 TI - Who benefits from nutritional support: what is the evidence? AB - Nutritional support, including oral nutritional supplements, enteral tube feeding and parenteral nutrition, can be an important part of the management of any patient. Malnutrition is common and costly, clinically and financially, if left untreated. In patients with, or at risk of, malnutrition, the appropriate use of nutritional support can prevent complications arising, produce other clinical, functional and financial benefits, and can be life saving in some situations. This article discusses the evidence from systematic reviews and meta-analyses of the effectiveness of nutritional support. PMID- 17413284 TI - Selecting the most appropriate route of feeding. AB - This article provides an overview of the various methods available for providing nutritional support. The various techniques available for both enteral and parenteral access are discussed. The selection of the most appropriate route of nutritional support is highly individual and recommendations are made regarding the factors that should be considered by the patient and the clinician in the decision-making process. PMID- 17413287 TI - Should patients with cancer be offered nutritional support: does the benefit outweigh the burden? AB - Nutrition support has been widely advocated as adjunctive therapy for a variety of underlying illnesses, including surgery and medical oncotherapy (radiation or chemotherapy for cancer). Both parenteral and enteral nutrition have been mistakenly viewed as feeding, when, in fact, they are medical interventions with associated risks and costs. The argument that nutrition support has to be provided to patients to prevent 'starving to death' confuses the difference between dying in a malnourished state and dying as a direct consequence of nutrient deprivation; cancer patients fit into the former category. As is true for any other medical intervention, efficacy is best established by randomized controlled clinical trials. When these forms of nutrition support have been so assessed, they have not usually been found to be any more efficacious than food on a tray or intravenous 5% dextrose solutions. In fact, parenteral nutrition actually caused harm in patients receiving medical oncotherapy (more total and infectious complications and fewer tumor responses). With regard to cancer patients, the only benefit that was demonstrated was the use of preoperative parenteral nutrition in patients undergoing attempted curative surgery for cancer of the upper gastrointestinal tract (esophagus, stomach, or pancreas). As nutrition support has associated complications (infections, mechanical problems with the tubes, and metabolic problems from the infusates) as well as costs, it cannot be recommended for cancer patients with the exception of the preoperative care of those with upper gastrointestinal malignancies and the occasional patient with gastrointestinal tract inadequacy owing to a slow-growing lesion. PMID- 17413288 TI - Nutritional support at the end of life: the relevant ethical issues. AB - This article attempts to summarize the ethics of nutritional support at the end of life. Although ethics are timeless, they have to be applied or adapted to new situations arising from our ability to prolong life by the application of relatively new nutritional treatments. The application of the law, and guidance from professional bodies on withholding or withdrawing treatment remains an emotive challenge for all involved in nutritional care and for society as a whole. PMID- 17413286 TI - The scientific rationale for optimizing nutritional support in cancer. AB - Cancer patients lose weight as a result of the anorexia-cachexia syndrome, and this weight loss is associated with significant morbidity and mortality. Thus, nutritional support to arrest or reverse weight loss is of paramount importance in the management of Cachexia cancer patients. Persistent tumour-induced metabolic changes result, however, in a suboptimal response to such support, making nutritional maintenance or improvement difficult targets to achieve. Mechanisms involved in the blockade to anabolism in cancer cachexia include alterations in skeletal muscle and hepatic protein metabolism, and reduced physical activity. Mediators underlying these mechanisms of weight loss include proinflammatory cytokines, tumour-specific cachectic factors, and neuroendocrine mediators of muscle catabolism. The complex mix of different mediators renders unimodal nutritional intervention a strategy that is unlikely to succeed completely. Therefore, clinical trials using combination therapies or immunonutrition are required for future success. PMID- 17413285 TI - Novel substrates. AB - Enteral and parenteral feeds need at least to contain adequate amounts of water, energy, protein, electrolytes, vitamins and trace elements. Ready-manufactured parenteral feeds for example are incomplete because of shelf-life constraints and require the addition of vitamins (especially) and trace elements. Acute vitamin deficiencies, notably thiamine deficiency, can be precipitated if this is not adhered to. An increasing interest, however, exists in the use of feeds containing substrates, which are intended to improve patient outcome in particular clinical circumstances. The purpose of this article is to examine as to what is available and make recommendations on their use. It deals with artificial feeds only - disease-specific diets are outside our remit. PMID- 17413289 TI - Nutritional support at the end of life: the relevant legal issues. PMID- 17413290 TI - Nicorandil-associated ulcerations. AB - This leading article refers to the paper by Abdelrazeq AS, Owen C, Smith L, McAdam JG, Pearson HJ, Leveson SH. Nicorandil-associated para-stomal ulceration: case series Eur J Gastroenterol Hepatol 2006; 18:1293-1295. We apologise to all concerned for the dissociation between the two papers, which was due to an administrative error. Nicorandil is used widely in patients with coronary artery disease. Nicorandil is well tolerated with only minor side effects. Nicorandil's association with oral, anal, gastrointestinal ulceration, and more recently para stomal ulceration has been reported. Medical awareness of nicorandil association with ulcerations should be high to help avoid unnecessary and harmful treatment as only cessation of the drug would heal the ulceration. Nicorandil is an antianginal drug used for the treatment of symptomatic coronary artery disease. It is characterized by an arterial and venous vasodilator effect with dual mechanism of action. Nicorandil is not a first-line agent in the management of angina but it is used in combination with other antianginal medications in stable and unstable angina. It is generally well tolerated with minor side effects such as headache, nausea, flushing and dizziness. The association of nicorandil with mouth and anal ulcers as well as the association with ulceration throughout the gastrointestinal tract has been reported, and recently, an association with para stomal ulceration has also been described. Medical awareness of the association of nicorandil with ulceration in any part of the gastrointestinal tract should be highlighted among all medical professionals to help avoid delays in withdrawing the treatment and to avoid unnecessary and sometimes invasive and costly interventions. PMID- 17413291 TI - Idiopathic megacolon. AB - This leading article refers to the paper by Meier-Ruge WA, Muller-Lobeck H, Stoss F, Bruder E. The pathogenesis of idiopathic megacolon. Eur J Gastroenterol Hepatol 2006; 18:1209-1215. We apologise to all concerned for the dissociation between the two papers, which was due to an administrative error. The pathogenesis of idiopathic megacolon is still unclear. Besides abnormalities of the enteric nervous system, alterations in the function of intestinal smooth muscle cells and connective tissue elements might play an important role. A permanent extension of the bowel diameter without concrete hints to its aetiology is termed idiopathic megacolon. Evidence exists that idiopathic megacolon comprises a heterogeneous group of conditions characterized by alterations of the enteric nervous system, smooth muscle cells and/or connective tissue. Innovative molecular techniques are needed to get further insights into the pathogenesis of these intestinal motility disorders. PMID- 17413292 TI - Influence of corticotropin-releasing hormone on gastric sensitivity and motor function in healthy volunteers. AB - BACKGROUND: As stress may be involved in the generation of functional dyspeptic symptoms, we evaluated the effect of the stress hormone, corticotropin-releasing hormone, on proximal stomach function. Twelve healthy volunteers [six women; 23 years (20-26 years)] underwent a barostat study on 2 days. During the infusion of corticotropin-releasing hormone (2.3 microg/kg/h) or saline, a stepwise distension protocol was performed followed by ingestion of a liquid meal (Nutridrink, 200 ml, 300 kcal). RESULTS: Corticotropin-releasing hormone infusion induced a significant increase in cortisol levels and basal volumes compared with placebo. The threshold for discomfort, meal-induced accommodation, dyspeptic symptoms, heart rate and blood pressure were all not significantly altered by corticotropin-releasing hormone infusion. CONCLUSION: In healthy volunteers, peripheral infusion of corticotropin-releasing hormone reduces basal fundic tone, but has no effect on meal-induced accommodation or visceral sensitivity to gastric distension. Our findings suggest that in healthy volunteers, peripheral corticotropin-releasing hormone seems not to be involved in the onset of dyspeptic symptoms. PMID- 17413293 TI - Risk factors for delayed postendoscopic mucosal resection hemorrhage in patients with gastric tumor. AB - OBJECTIVE: Although endoscopic mucosal resection has been recognized as the standard treatment for gastric mucosal neoplasm, postendoscopic mucosal resection hemorrhage remains a major complication of endoscopic mucosal resection, and this problem seems to be increasing owing to the development of invasive techniques. The aims of this study were to determine the incidence and grade of postendoscopic mucosal resection hemorrhage and to identify risk factors for delayed postendoscopic mucosal resection hemorrhage in patients with gastric neoplasm. METHODS: Data of endoscopic mucosal resections performed by three endoscopists were retrospectively collected over 8 years and then analyzed. Immediate postendoscopic mucosal resection hemorrhage was defined as bleeding during the procedure. Delayed postendoscopic mucosal resection hemorrhage was defined when two of the four following parameters were satisfied after the endoscopic mucosal resection period; (i) hematemesis, melena or dizziness, (ii) hemoglobin loss >2 g/dl, (iii) blood pressure decrease >20 mmHg or pulse rate increase >20/min and (iv) Forrest I or IIa-IIb on follow-up endoscopy. RESULTS: A total of 157 patients (mean age: 64 years, male : female=44 : 113) were reviewed. Twenty-nine (18.5%) and 13 patients (8.3%) presented with immediate and delayed postendoscopic mucosal resection hemorrhage, respectively. Multivariate logistic regression analysis revealed that the patient's age (15 mm; odds ratio 5.90, 95% confidence interval 1.13-30.87) and the experience of the endoscopist (or=3 mm; thrombotic occlusion or angiographic evidence of thrombus; absence of severe proximal tortuosity or calcification. We evaluate the efficacy of TA in terms of procedural success, coronary thrombolysis in myocardial infarction (TIMI) flow, myocardial blush grade (MBG), resolution>or=50% of ST segment elevation, and clinical events during hospital stay and at 6-month follow up. RESULTS: A total of 486 primary PTCAs were performed, 217 (44.6%) with TA as a first device using RESCUE (n=65), EXPORT (n=140) and DIVER-CE (n=12) catheters. In 141 (65%) cases, macroscopic material was aspirated. The patients submitted to TA were more often males (84.7% versus 71.7%, P<0.05) and younger (age: 61.02+/ 11.91 versus 64.47+/-10.59 years, P<0.01) than patients treated with traditional PTCA and the IRA was more frequently occluded at angiography (basal TIMI 0: 70.5% versus 47.9%). Application of the TA did not increase the complexity of the procedure (door-to-balloon times, minutes of fluoroscopy and amount of dye). TA alone was effective to restore TIMI 3 flow in 187 cases (86.2%) as a first device and in three other cases (1.4%) after predilatation with balloon. Direct stenting without predilatation was possible in 144 cases (66.4%) after TA. TA was not effective in 27 cases (12.4%) and this subgroup had both angiographic and clinical unfavourable results in comparison with the effective TA group (final TIMI 1 in 11.1% versus 0.5%, P<0.015; final MBG 1 in 55.5% versus 9.5%, P<0.001; lack of ST segment resolution>or=50% in 44.4% versus 7.9%, P<0.001; in-hospital mortality 14.8% versus 2.6%, P<0.05 and mortality at 6 months 18.5% versus 3.1%, P<0.05). In the whole TA population, final TIMI 3 flow was achieved in 203 cases (93.5%), final MBG 3 in 145 cases (66.8%) and ST segment resolution>or=50% in 185 cases (85.2%), in-hospital mortality was 4.1% and cumulative mortality at 6-month follow-up was 5.5%. CONCLUSIONS: In our case series, 486 consecutive unselected patients with STEMI were treated in a primary PTCA high-volume centre using TA devices. Our study demonstrates that, in STEMI patients treated with primary PTCA, a routine strategy with TA before angioplasty guided by angiographic selection criteria is feasible in almost 50% of cases, is safe and effective, does not increase procedural time and offers good results in terms of tissue perfusion, both epicardial (TIMI flow) and myocardial (MBG, ST regression). When successfully performed, TA identifies a population with favourable in-hospital and 6-month outcome. PMID- 17413303 TI - Real-time three-dimensional echocardiography: a pilot feasibility study in an Italian cardiologic center. AB - BACKGROUND: The majority of studies demonstrating the diagnostic potential of three-dimensional (3-D) echocardiography have been conducted on selected series of patients in research laboratories. AIM: To investigate the feasibility and usefulness of real-time 3-D transthoracic echocardiography in daily routine practice. METHODS: Two hundred consecutive patients underwent standard two dimensional (2-D) transthoracic echocardiography (TTE) and real-time (RT) 3-D TTE with a commercially available ultrasound system (Sonos 7500 LIVE 3D, Philips Medical Systems). The quality of 3-D acquisitions and post-processed images was graded as: bad, satisfactory, good and demo. In each case, the results of 3-D TTE were compared with 2-D images to disclose additional qualitative information provided by 3-D examination. An additional qualitative information score was given for each cardiac structure. RESULTS: The mean time of the 3-D examination was 11+/-4 min. The mean time of 2-D transthoracic studies in our laboratory is 25 min and the total time in this series was therefore approximately 36 min. The mean number of acquisitions in our series was 11.5 per patient. The quality was evaluated as bad/insufficient in 7.0%, satisfactory/sufficient in 29.6%, good in 40.2% and demo in 23.2% of all datasets and reconstructions. The structures with greater additional qualitative information scores comprise the anterior and posterior mitralic leaflets, antero-lateral and postero-medial papillary muscles and leaflets of tricuspid valve. The intra- and interobserver reproducibility of quality grading was good and there are few interobserver discrepancies, which were resolved by two physicians, experienced in 3-D echocardiography, not involved in the study. CONCLUSIONS: RT 3-D TTE may be used in clinical settings with high feasibility rate and may provide additional, clinically quite relevant qualitative information. This technique may expand the abilities of non-invasive cardiology and open new doors for the evaluation of cardiac disease. PMID- 17413304 TI - Transthoracic echocardiography-guided biopsy of a right ventricular mass. AB - Biopsy of the heart has become a widely applied technique in circumstances such as detection of a transplant reject or histological diagnosis of masses. Biopsy is usually performed in the catheter laboratory (cath-lab) under fluoroscopic guidance. Echocardiography guidance is employed but as 'ancillary' comforting support to the cath-lab operator and for the early detection of complications. Until now, only a transesophageal approach has been used for guiding biopsy of intracardiac masses. Today, thanks to the current echocardiographic technology employing second harmonics, the definition of cardiac structures has consistently improved in the transthoracic approach, possibly allowing the full transthoracic procedure to be performed. We describe the biopsy of a large mass in the right ventricle, completely conducted without the help of fluoroscopy, and completed only under the help and guidance of transthoracic echocardiography. PMID- 17413305 TI - Non-obstructive prosthetic aortic valve thrombosis presenting with acute myocardial infarction and stroke in a patient with inadequate low-molecular weight heparin treatment. The unlucky patient and the (pseudo)prudent physician. AB - Non-obstructive prosthetic valve thrombosis is a rare and underestimated complication in patients with left-sided mechanical heart valves. Systemic embolisation, mainly involving the cerebral circulation, often represents the first clinical manifestation. We report a case of multiple, successive embolizations in the coronary and cerebral circulation, presenting with an acute myocardial infarction and stroke in a patient with latent, non-obstructive thrombosis of a mechanical bileaflet aortic valve. Because of scheduled urological surgery, chronic vitamin K antagonist treatment had previously been discontinued and replaced with low-molecular-weight heparin, at inadequate dosage. Following coronary arteriography, brain computed tomography scan and transoesophageal echocardiography, thrombolysis was performed successfully. This case emphasises the utility of performing transoesophageal echocardiography routinely in the presence of ischaemic signs in patients with mechanical heart valves. In patients requiring discontinuation of oral anticoagulant therapy, accurate management and continuous monitoring of alternative medications are needed in order to avoid severe thromboembolic complications. PMID- 17413306 TI - An unusual case of tricuspid lesion in congenital corrected transposition of the great arteries. AB - An unusual case of severe supravalvular stenosing ring of the left atrium associated with tricuspid valve dysplasia in an adult symptomatic patient affected by congenitally corrected transposition and unrestrictive ventricular septal defect is reported. The stenosis of the systemic atrioventricular valve possibly prevented the development of obstructive pulmonary vascular disease; removal of the membrane, attached to the tricuspid annulus, together with pulmonary banding was carried out. Clinical conditions improved after surgery. The unusual stenotic lesion of the systemic right ventricular inflow allowed conservative surgical palliation in this adult patient. PMID- 17413307 TI - Late coronary thrombosis in paclitaxel-eluting stents. Case reports. AB - Drug-eluting stent (DES) implantation has reduced angiographic and clinical restenosis that actually develops in less than 10% of treated patients. DESs also tend to delay the endothelialisation process increasing the risk of stent thrombosis. Subacute stent thrombosis may complicate long-term success of coronary angioplasty; it is a sudden event and usually causes acute myocardial infarction or sudden cardiac death. Patients undergoing DES implantation should be treated with dual antiplatelet therapy for at least 3-6 months. We describe two cases presenting with ST-elevation acute myocardial infarction due to stent thrombosis that occurred late after deployment of a paclitaxel-eluting stent, after discontinuation of antiplatelet therapy. It is important, for clinical cardiologists and general practitioners, to know the potential risk of late thrombosis of DES patients and, consequently, the implications regarding management of antiplatelet therapy. PMID- 17413308 TI - Pseudocoarctation as cause of a refractory hypertension. AB - Pseudocoarctation of the aorta is an abnormality of the descending portion of the aortic arch at the level of the isthmus without a real pressure gradient. We report a case of a stenotic pseudocoarctation causing severe refractory hypertension. PMID- 17413309 TI - Unusual presentation of ruptured descending thoracic aortic aneurysm. AB - We here report a very unusual presentation of ruptured thoracic aortic aneurysm. In a 50-year-old patient, almost asymptomatic and in stable clinical conditions, the chest radiograph and computed tomography scan revealed a right-sided rupture of a previously undiagnosed thoracic aortic aneurysm. The patient was treated successfully with an emergency surgical procedure. PMID- 17413310 TI - Prognostic role of post-infarction C-reactive protein in patients undergoing implantation of cardioverter-defibrillators: design of the C-reactive protein Assessment after Myocardial Infarction to GUide Implantation of DEfibrillator (CAMI GUIDE) study. AB - BACKGROUND: Patients at risk of sudden cardiac death (SCD) after myocardial infarction (MI) can currently be offered effective means of prevention, such as implantable cardioverter-defibrillators (ICD). However, predictors of SCD able to identify those patients who are at higher risk are still lacking. Whether C reactive protein (CRP), a serum inflammatory marker with established prognostic accuracy after MI, can also be a predictor of SCD is unclear. METHODS: The CAMI GUIDE study is designed to evaluate the prognostic role of CRP in patients undergoing ICD implantation after MI according to MADIT II criteria (i.e. left ventricular ejection fractionT (p.Gln380X), c.2160_ 2161insC (p.Cys721fs), and c.1707C>G (p.Ser569Arg). Patients were either homozygous for one of these mutations or compound heterozygotes, and they differed in their phenotype. CONCLUSION: We report the potential pitfalls of performing homozygosity mapping in a highly consanguineous population and discuss the phenomenon of multiple mutations in a given gene within an isolate. PMID- 17413423 TI - Genetic counseling in adult carriers of a balanced chromosomal rearrangement ascertained in childhood: experiences from a nationwide reexamination of translocation carriers. AB - PURPOSE: Prenatal diagnosis is offered to carriers of a balanced chromosomal rearrangement because it may predispose to offspring with an unbalanced karyotype. Therefore, carriers examined prenatally or in childhood should be informed before they reach reproductive age. We aimed to determine how many of the adult carriers ascertained in childhood currently know about their carrier status. METHODS: We used data obtained by a questionnaire study reexamining carriers of a balanced reciprocal translocation. When a carrier was older than 18 years of age and had been examined in childhood, relatives were asked whether she/he knew of the translocation. RESULTS: Among the 113 parents we interviewed, 10 carriers (9%) in 8 families had not been informed. In one of the eight families, an offspring with an unbalanced translocation was born 23 years after the father had been examined in childhood. CONCLUSION: Because of our findings, the practice of genetic counseling in Denmark has been changed: When a carrier of a balanced chromosomal rearrangement who was examined prenatally or in childhood turns 18 years of age, the parents will receive a letter reminding the family about the reproductive risk. PMID- 17413424 TI - Prenatal/neonatal pathology in two cases of Cornelia de Lange syndrome harboring novel mutations of NIPBL. AB - PURPOSE: This study reviews prenatal findings in two cases with a suspected diagnosis of Cornelia de Lange Syndrome, a multisystem disorder characterized by somatic defects and mental retardation, that were later confirmed by postmortem examination and molecular testing. Although the correlation between the Cornelia de Lange Syndrome genotype and phenotype is still unclear, preliminary data indicate several severe phenotypic features that are likely to be detected prenatally in NIPBL-mutated patients. METHODS: We report on two prenatal/neonatal cases with unusual pathologic findings indicating Cornelia de Lange Syndrome. The first, with suspected Cornelia de Lange Syndrome after a set of typical dysmorphisms was noted by prenatal ultrasound, was confirmed by a physical examination after termination of the pregnancy. The second was diagnosed neonatally on the basis of typical clinical signs. Medical complications led to death within the first month of life. RESULTS: Molecular analysis of NIPBL, the gene that codes for delangin (a component of the cohesin complex), performed postnatally detected two de novo mutations: a missense change (P2056L) in a highly conserved residue and a nonsense alteration (S2490 replaced by a stop codon). CONCLUSION: We suggest that early diagnosis of Cornelia de Lange Syndrome would be made much easier by the assemblage of a set of prenatal diagnostic features and criteria in Cornelia de Lange Syndrome cases that have been confirmed by direct physical and molecular examinations. We also suggest that Cornelia de Lange Syndrome genotype-phenotype correlations need to be extended to prenatal cases. PMID- 17413426 TI - Emergency department evaluation of ventricular shunt malfunction: is the shunt series really necessary? AB - OBJECTIVE: The malfunction of a ventricular shunt is one of the most common clinical problems encountered in pediatric neurosurgery. Standard emergency department (ED) evaluation of suspected shunt malfunction consists of plain radiographs of the skull, neck, chest, and abdomen (shunt series) to look for mechanical breaks, kinks, and disconnections in the shunt, and a cranial computed tomography (CT) scan to evaluate for signs of increased ventricular size. We hypothesized, however, that in the context of a cranial CT scan that did not demonstrate a shunt malfunction, obtaining the shunt series would not prove to be clinically useful. METHODS: A retrospective chart review was conducted of all patients younger than 18 years with a history of a ventricular shunt who presented to an urban, tertiary pediatric ED between January 1, 2000, and September 30, 2004, for suspected shunt malfunction. Demographic and clinical characteristics of patients were recorded, as well as the results of shunt series and cranial CT scans. Shunt malfunction was defined as the performance of a shunt revision within 1 week of radiographic evaluation. RESULTS: During the study period, 291 children with a ventricular shunt were evaluated in the ED 461 times for suspected shunt malfunction. The mean age of patients was 90.6 months (SD, 71.5 months); 163 (58.5%) were men, and 209 (71.8%) were white. Three hundred sixty patients (78.1%) had a shunt series performed during their ED evaluation, and 410 (88.9%) had a CT scan of the head. Seventy-one patients (15.4%) were diagnosed with shunt malfunction. Twenty-two had a normal cranial CT scan. Of these patients, 6 had an abnormal shunt series, and 14 had a normal shunt series. CONCLUSIONS: The routine use of the shunt series seems warranted in the evaluation of the child with suspected shunt malfunction as children with shunt malfunction may present with a normal cranial CT scan but an abnormal shunt series. PMID- 17413427 TI - The increasing use of pediatric emergency facilities in the evening. AB - OBJECTIVES: In Japan, the demand for pediatric emergency medicine has been increasing, especially in the evening. The purpose of this study was to identify the reasons for overcrowding of pediatric emergency facilities in the evening. METHODS: A population-based survey was conducted in Yokohama City, Japan, that targeted parents of children 1 or 3 years of age. These children participate in regular health checkups. Questionnaires about their child's illnesses and the pediatric emergency system were mailed to 30,000 parents of a child expected to undergo a health checkup between May 2004 and January 2005. RESULTS: Data obtained from the completed questionnaires indicated that many parents noticed their child's illness or injury most frequently during the evening, when most medical facilities are closed. The peak period when parents noticed their child's illness was the evening (4:00 pm-12 midnight, 54.4%), followed by the daytime (8:00 am-4:00 pm, 30.3%) and then the nighttime (12 midnight-8:00 am, 15.3%). During all 3 periods, parents felt it difficult to judge their child's condition and thus many used emergency facilities unnecessarily. CONCLUSIONS: The overcrowding of pediatric emergency facilities in the evening is likely due mainly to a mismatch between the peak time of children's illnesses and the office hours of pediatric clinics. Parents' difficulties in assessing their child's condition and anxiety over their child's illness and injuries seem to be other factors that contribute to this imbalance. PMID- 17413428 TI - Prehospital intranasal midazolam for the treatment of pediatric seizures. AB - BACKGROUND: The local emergency medical services (EMS) council implemented a new pediatric treatment protocol using a Mucosal Atomization Device (MAD) to deliver intranasal (IN) midazolam for seizure activity. METHODS: We sought to compare outcomes in seizing pediatric patients treated with IN midazolam using a MAD (IN MAD midazolam) to those treated with rectal (PR) diazepam, 18 months before and after the implementation of the protocol. RESULTS: Of 857 seizure patients brought by EMS to our emergency department (ED), 124 patients (14%) had seizure activity in the presence of EMS and were eligible for inclusion in this study. Of the 124 patients eligible for this study, 67 patients (54%) received no medications in the prehospital setting, 39 patients (32%) were treated with IN MAD midazolam, and 18 patients (15%) were treated with PR diazepam. Median seizure time noted by EMS was 19 minutes longer for PR diazepam (30 minutes) when compared with IN-MAD midazolam (11 minutes, P = 0.003). Patients treated with PR diazepam in the prehospital setting were significantly more likely to have a seizure in the ED (odds ratio [OR], 8.4; confidence interval [CI], 1.6-43.7), ED intubation (OR, 12.2; CI, 2.0-75.4), seizure medications in the ED to treat ongoing seizure activity (OR, 12.1; CI, 2.2-67.8), admission to the hospital (OR, 29.3; CI, 3.0-288.6), and admission to the pediatric intensive care unit (OR, 53.5; CI, 2.7-1046.8). CONCLUSIONS: The IN-MAD midazolam controlled seizures better than PR diazepam in the prehospital setting and resulted in fewer respiratory complications and fewer admissions. PMID- 17413429 TI - Use of decubitus radiographs in the diagnosis of foreign body aspiration in young children. AB - BACKGROUND: Lateral decubitus radiographs are often obtained in young children with suspected foreign body aspiration. Their usefulness has not been well studied. OBJECTIVES: To assess the value of decubitus radiographs in detecting foreign body aspiration in young children and compare their value to history and physical examination. METHODS: Retrospective review of children younger than 4 years with suspected foreign body aspiration who had decubitus radiographs and underwent bronchoscopy over a 5-year period. Patients with proven foreign body aspiration were compared with those without foreign bodies for differences in symptoms, signs, location and character of the foreign body, and the diagnostic value of standard and decubitus chest radiographs. RESULTS: Twenty-eight of 41 children who underwent bronchoscopy for possible foreign body aspiration had decubitus radiographs. Foreign bodies were identified in 22 patients (79%). A total of 27% of children with foreign body aspiration and 33% of children without a foreign body had suggestive decubitus radiographs (P = not significant). As a measure of detecting foreign body aspiration, positive decubitus radiographs had a sensitivity of 27%, a specificity of 67%, a positive predictive value of 75%, and a negative predictive value of 20%. The odds ratio of finding a foreign body with suggestive decubitus radiographs was 0.75 (95% confidence interval [CI], 0.1 5.2; P = 0.57). Foreign body aspiration was confirmed in 94% of children if there was both a sudden onset of symptoms and a witnessed choking episode (odds ratio, 13.3; 95% confidence interval, 1.3-138.9; P = 0.02). CONCLUSION: Decubitus chest radiographs, at least as routinely performed and interpreted, seem to add little to the evaluation of young children with suspected foreign body aspiration. A history of a witnessed choking episode combined with a sudden onset of respiratory symptoms remains the most important indication for bronchoscopy. PMID- 17413431 TI - Low-dose ketamine: efficacy in pediatric sedation. AB - OBJECTIVES: Intravenous (IV) ketamine has gained widespread use in the emergency department (ED) for procedural sedation. The most commonly recommended starting dose is 1.5 mg/kg. We examined whether lower doses of ketamine in the range of 0.5 to 1.0 mg/kg could successfully sedate pediatric patients. METHODS: We retrospectively reviewed quality assurance data of patients sedated with IV ketamine in a pediatric ED. Patients were administered 0.02 mg/kg of IV atropine, 0.05 mg/kg of IV midazolam, and then 0.5 mg/kg of IV ketamine. Additional aliquots of 0.25 to 0.5 mg/kg of ketamine were given as necessary, to a maximum of 2.0 mg/kg. Efficacy of sedation was assessed after every dose by pediatric emergency medicine attendings or fellows. RESULTS: Seventy-two patients had quality assurance forms completed. The total ketamine dose administered ranged from 0.5 to 2 mg/kg. Adequate procedural sedation was obtained for 70 (97%) of 72 patients. Forty-four percent of patients required 0.75 mg/kg or less of ketamine to obtain adequate initial sedation; 25% of subjects required only 0.5 mg/kg; 43% of patients required 1.0 mg/kg of IV ketamine. We found that 88% of our patients could be successfully sedated at initial doses of 1 mg/kg or less. CONCLUSIONS: Our study suggests a potential role for low-dose IV ketamine in the range of 0.5 to 1.0 mg/kg for pediatric procedural sedation. Most pediatric ED patients can be successfully sedated with 1 mg/kg of ketamine. PMID- 17413432 TI - Cardiac tamponade as a delayed presentation of Neisseria meningitidis infection in a 5-month-old infant. AB - This is a case of a 5-month-old female infant diagnosed with primary meningococcal pericarditis. Pericarditis is a well-recognized but uncommon complication of meningococcal infection. Primary meningococcal pericarditis, defined as purulent pericarditis without any clinical evidence of disseminated meningococcemia, meningitis, or other foci of meningococcal infection, is exceedingly rare, with only 21 reported cases since the first case was reported in 1939. This case report of primary meningococcal pericarditis is the youngest case and only the second case reported in an infant in the English literature to date. PMID- 17413433 TI - Cerebral infarct secondary to traumatic carotid artery dissection. AB - Internal carotid artery dissection is an important cause of ischemic stroke in children and young patients. Children presenting with gross neurological abnormalities after blunt trauma to the head or neck should be considered to have sustained injury to the carotid arteries until proven otherwise. Treatment options include observation, anticoagulation and endovascular stenting, and aggressive surgical repair of the carotid artery injury. We present the case of a 7-year-old boy who had a dissection of his right internal carotid artery after a dangerous position of head upside down from a water slide. PMID- 17413434 TI - A genital hair tourniquet in a 9-year-old girl. AB - The hair tourniquet is a well-described phenomenon. Typically, a hair or filament becomes tightly wrapped around an appendage, thereby causing swelling, pain, and, in extreme cases, necrosis. Affected areas include fingers, toes, and the genitalia. A case of a 9-year-old girl with a clitoral hair tourniquet is described, and a review of the literature of genital tourniquets in females is presented, with a discussion about potential etiology. The hair-thread tourniquet syndrome is a disease in which a hair or filament becomes wrapped around an end perfusion appendage, thereby limiting adequate venous and lymphatic drainage of that tissue. This results in hyperemia, swelling and pain. If uncorrected, edema and interstitial pressures may increase to prevent adequate vascular supply, and the affected area can progress to necrosis. We are classically taught that this syndrome primarily affects the fingers, toes, and infantile penis and is an important consideration in the differential diagnosis of the colicky infant. The author presents a case of a prepubescent girl with a genital hair tourniquet. PMID- 17413435 TI - Bilateral idiopathic vocal cord palsy. AB - We present 3 cases of bilateral vocal cord palsy who presented with acute respiratory distress with features of upper airway obstruction requiring tracheostomy. No cause could be found despite clinical evaluation and laboratory investigations. This diagnosis should be considered when child presents with upper airway obstruction emergency after ruling out other important causes of stridor and laryngoscopic examination is warranted in such cases for diagnosis. PMID- 17413436 TI - Do not laparotomize food-protein-induced enterocolitis syndrome. AB - Food protein-induced enterocolitis syndrome is a severe cell-mediated, non immunoglobulin E-mediated gastrointestinal food allergy, typically to milk and soy. It may masquerade as more common pediatric presentations, where the temporal relationship between symptomatology and exposure to food offers a vital clue to the underlying pathology. A case of food protein-induced enterocolitis syndrome presenting with features similar to intussusception, leading to unnecessary surgery, is presented. PMID- 17413437 TI - Emergency department management of the pediatric patient with supraventricular tachycardia. AB - Supraventricular tachycardia (SVT) is the most common tachyarrhythmia that necessitates treatment in children. It is characterized by a rapid and regular heart rate, which generally exceeds 180 beats per minute in children and 220 beats per minute in adolescents. Supraventricular tachycardia results from conduction of electrical impulses along an accessory connection from the atrium to the ventricle (atrioventricular reentry tachycardias: orthodromic or antidromic) or conduction within the atrioventricular node (atrioventricular node reentry tachycardia). Emergency department management of SVT depends on the patient's clinical status. Treatment of a stable patient with SVT includes vagal maneuvers and adenosine, whereas treatment of an unstable patient requires synchronized cardioversion. This article presents an overview of the etiology, pathophysiology, and clinical presentation of SVT and discusses the emergency department management of an infant or child with SVT. PMID- 17413438 TI - Pediatric emergency medicine: legal briefs. PMID- 17413439 TI - ECGs in the ED. PMID- 17413440 TI - A cyanotic toddler. PMID- 17413441 TI - My infant will not move the left arm. PMID- 17413442 TI - Ultrasound guidance for central vascular access in the pediatric emergency department. AB - Central vascular access is sometimes required for hemodynamic monitoring and infusion of fluids and medications in the pediatric emergency department. In many cases, it is attempted after failed peripheral venous and intraosseous access. Some evidence exists demonstrating benefits of ultrasound (US)-guided central vascular cannulation in adults in emergency departments. With appropriate education in its use, US-guided cannulation of central veins in children is likely to be associated with less complications and greater success. In the pediatric emergency department, the femoral vein is the most practical central venous cannulation site. A sound educational and quality assurance program is necessary for US-guided cannulation in the pediatric emergency department. PMID- 17413443 TI - Association between CCK-AR gene and schizophrenia with auditory hallucinations. AB - OBJECTIVE: Previous studies on a possible association between CCK-AR polymorphisms and schizophrenia have been controversial. The aim of the present study was to assess a potential association between schizophrenic patients with auditory hallucinations and polymorphisms of the CCK-AR gene. METHODS: A set of single nucleotide polymorphisms mainly located in the regulatory region of the CCK-AR gene was analysed in a sample of 163 Diagnostic and statistical manual of mental disorders-IV-diagnosed schizophrenic patients and 162 healthy controls. RESULTS: Significant differences in the genotype (P=0.011) and allele (P=0.0009) frequencies of the +121C/G SNP (located in the 5' regulatory region) were found between patients and controls. The excess of the C allele in the patient group remained significant after Bonferroni correction (P=0.03). However, functional in vitro assays, did not reveal significant differences on gene expression between +121G and +121C alleles of this SNP. Further investigations revealed two risk haplotypes: +121C/+978A/+984T (P=0.01) and +121C/+978T/+984C (P=0.0091) as well as a protective haplotype: +121G/+978T/+984T (P=0.0001). CONCLUSION: Our data support a possible role of the CCK-AR gene in the vulnerability to schizophrenia in patients with auditory hallucinations, and suggest remarkable allele heterogeneity. PMID- 17413444 TI - NCAM1 association study of bipolar disorder and schizophrenia: polymorphisms and alternatively spliced isoforms lead to similarities and differences. AB - OBJECTIVE: The neural cell adhesion molecule (NCAM1) is a multifunction transmembrane protein involved in synaptic plasticity, neurodevelopment, and neurogenesis. Multiple NCAM1 proteins were differentially altered in bipolar disorder and schizophrenia. Single nucleotide polymorphisms (SNPs) in the NCAM1 gene were significantly associated with bipolar disorder in the Japanese population. Bipolar disorder and schizophrenia may share common vulnerability or susceptibility risk factors for shared features in each disorder. METHODS: Both SNPs and splice variants in the NCAM1 gene were analysed in bipolar disorder and schizophrenia. A case-control study design for association of SNPs and differential exon expression in the NCAM1 gene was used. RESULTS: A genotypic association between bipolar disorder and SNP b (rs2303377 near mini-exon b) and a suggestive association between schizophrenia and SNP 9 (rs646558) were found. Three of the two marker haplotypes for SNP 9 and SNP b showed varying frequencies between bipolar and controls (P<0.0001) as well as between schizophrenia and controls (P<0.0001). There were nine NCAM1 transcripts present in postmortem brain samples that involve alternative splicing of NCAM1 mini-exons (a, b, c) and the secreted (SEC) exon. Significant differences in the amounts of four alternatively spliced isoforms were found between NCAM1 SNP genotypes. In exploratory analysis, the c-SEC alternative spliced isoform was significantly decreased in bipolar disorder compared to controls for NCAM1 SNP b heterozygotes (P=0.013). CONCLUSIONS: Diverse NCAM1 transcripts were found with possibly different functions. The results suggest that SNPs within NCAM1 contribute differential risk for both bipolar disorder and schizophrenia possibly by alternative splicing of the gene. PMID- 17413445 TI - Brain-derived neurotrophic factor gene polymorphisms and schizophrenia: a meta analysis. AB - OBJECTIVES: To evaluate whether the G196A and C270T polymorphisms of the brain derived neurotrophic factor gene are associated with increased risk of schizophrenia. METHODS: A meta-analysis of nine genetic association studies was carried out. The meta-analysis included genotype data on 1404/1597 schizophrenics/controls for G196A and 877/989 schizophrenics/controls for C270T. RESULTS: The overall analysis for investigating the association of the G196A allele G and the risk of developing schizophrenia relative to the allele A, showed significant evidence of heterogeneity (P=0.05, I(2)=58%) between the studies and nonsignificant association [random effects odds ratio 1.08 and 95% confidence interval (0.88-1.32)]. In Caucasians, there was a trend towards heterogeneity (P=0.19, I(2)=40%), then, the random and fixed effects odds ratios were 1.24 (0.96-1.60) and 1.27 (1.06-1.53), respectively. For the C270T polymorphism, overall, there was significant evidence of heterogeneity between studies (P=0.07, I(2)=55%) and the allele T was associated with a 63% increased risk of schizophrenia compared with C allele [random effects odds ratio 1.63 (1.01-2.65)]. The dominant model for allele T produced significant association [random effects odds ratio 1.68 (1.02-2.79)]. No source of bias was seen in the selected studies and the differential magnitude of effect in large versus small studies was not significant. CONCLUSIONS: The meta-analysis results provided a weak evidence of association between C270T polymorphism and schizophrenia, and large heterogeneity between studies, whereas there was no evidence of association for G196A polymorphism. The above findings reinforce the need for large and more rigorous association studies. PMID- 17413446 TI - Further genetic characterization of the fawn-hooded (FH/Wjd) rat, an animal model of comorbid depression and alcoholism. AB - OBJECTIVE: The main objective of this study was a more detailed genetic characterization of the alcohol preferring fawn-hooded rat and its intercrosses. Fawn-hooded rats drink substantially more alcohol voluntarily than the ACI rats. The fawn-hooded rats were shown to be more immobile in the forced-swimming test and to drink more saccharin. Recent comparisons of the parental strains with F1 and F2 intercrosses revealed that the alcohol and saccharin intakes were positively correlated with each other but not with immobility. METHODS: The F1 and F2 progeny were generated by intercrossing the fawn-hooded and ACI/N rats. Data from the F2 progeny, their F1 parents and progenitors were used to estimate heritability. RESULTS: Heritability was estimated for alcohol intake (75.6% in males and 67.1% in females), alcohol preference (64.7% in males and 39.2% in females), saccharin intake (50.8% in males and 37.5% in females), and immobility (50.2% in males and 72.1% in females). This same data provided estimates of the number of genes involved in these phenotypes between three and six. We also took advantage of the fact that both progenitor strains are pigmented, so a tremendous variety of coat colors were present in the F2 progeny (i.e. black, black-hooded, agouti, agouti-hooded, fawn, fawn-hooded, orange, and orange-hooded). Coat color analyses indicated that none of the variables significantly varied with coat color. A high correlation however, was observed between alcohol intake and preference in each group. Significant correlations between alcohol and saccharin intakes were seen only in some groups. CONCLUSION: These findings suggest that these phenotypes might be regulated by multiple genes, which could be detected in quantitative trait loci. These analyses are currently underway and will provide a novel approach in understanding the genetics of voluntary alcohol drinking. PMID- 17413447 TI - Adult metachromatic leukodystrophy: a new mutation in the schizophrenia-like phenotype with early neurological signs. AB - OBJECTIVES: The adult type of metachromatic leukodystrophy can manifest itself as motor or as psycho-cognitive form, the latter is very similar to schizophrenia. We report on two sisters with adult metachromatic leukodystrophy who display symptoms of both forms. METHODS: Presented are genotype analyses and 4-year follow-up data regarding clinical manifestations as well as neurocognitive and neuroimaging results for two adult sisters with metachromatic leukodystrophy. RESULTS: Whereas the younger sister developed disorganized schizophrenia-like symptoms, the other exhibited schizophrenia-like, negative symptoms. In both sisters, neurological signs were already present at the onset of the disease and progression towards dementia was documented within 1-2 years. In peripheral leukocytes, the activity of arylsulphatase A was reduced to 2 and 5% of the mean normal activity in both women. Genotype analysis revealed compound heterozygosity for a known severe splice site mutation, (c.459+1G>A) together with two known polymorphisms, [(c.937G>T), (p.Trp193Asp)] and [(c.1530C>G), (p.Thr391Ser)], and a novel missense mutation, (c.1194C>T). The latter results in the exchange of a conserved polar amino acid, threonine 279, to hydrophobic isoleucine (Thr279Ileu), which could not be found among >100 control alleles. A family analysis identified T279I as the paternal allele, whereas (c.459+1G>A) as well as the two polymorphisms were inherited from the mother. This is consistent with a disease-causing effect of the novel mutation. CONCLUSIONS: The novel mutation, T279I detected in our patients, correlates with a specific phenotype with schizophrenia-like symptoms, neurological signs and cognitive impairment early in the course of the disease and a relatively fast progression towards dementia. This is in contrast to previous reports on adult metachromatic leukodystrophy patients with the psycho-cognitive phenotype who did not show any neurological signs for decades, however, most of these patients were heterozygous for another specific missense mutation, I179S. PMID- 17413448 TI - Parent-of-origin effect in the segregation analysis of bipolar affective disorder families. AB - OBJECTIVE: The purpose of this study is to test for heterogeneity in bipolar families based on the differential parental transmission of disease. METHODS: Complex segregation analyses of 260 bipolar families, ascertained by the Johns Hopkins Bipolar Disorder Study, was performed based on the evidence for a parent of-origin effect in the inheritance pattern by using REGD in Statistical Analysis for Genetic Epidemiology, Release 3.1 program. RESULTS: A Mendelian dominant model provided the best explanation in 57 paternal pedigrees (pedigrees with an affected paternal lineage). No evidence of Mendelian inheritance existed among 141 pedigrees showing maternal transmission. A likelihood ratio test for heterogeneity on the basis of best-fitting Mendelian dominant model showed significant differences between these two groups. CONCLUSION: These results suggest that pedigrees with no evidence of maternal transmission of bipolar disorder may represent a unique genetic subgroup of multiplex bipolar families. PMID- 17413449 TI - Familial association of schizophrenia symptoms retrospectively measured on a lifetime basis. AB - OBJECTIVE: The objective of this study was to characterize the familial associations of symptoms or symptom dimensions of schizophrenia, as a component of the effort to delineate suitable phenotypic markers under the homogenous genetic control. METHODS: Clinical symptoms of the patients were evaluated on a lifetime basis, with the Korean version of the Diagnostic Interview for Genetic Studies and the Krawieka scale. Our sample consisted of 54 sibling pairs who were concordant for Diagnostic and statistical manual of mental disorder-IV schizophrenia or schizoaffective disorder. RESULTS: At the individual symptom level measured by the Diagnostic Interview for Genetic Studies, we observed a statistically significant degree of concordance within the sibling pairs for auditory hallucination (kappa=0.600, chi(2)=15.193, P=0.000). The within-sibling resemblance of this symptom was also indicated by the results of correlation analyses of individual items of the Krawiecka scale (partial correlation coefficient=0.423, P=0.005). Among the individual symptoms or symptom dimensions of the Krawiecka scale, the 'anxious' (partial correlation coefficient=0.469, P=0.001) and the 'affective dimension' (including the items of 'anxious' and 'depressed') (partial correlation coefficient=0.436, P=0.003) were also determined to indicate significant intra-pair correlations after controlling for potential confounding variables, such as sex, duration of illness, and age of onset. CONCLUSIONS: The results of this study suggest that familial factors, possibly genetic factors, contribute a liability to the development of auditory hallucinations. This symptom might then prove useful as a phenotype marker in future genetic studies. PMID- 17413450 TI - The gene for synapsin III and attention-deficit hyperactivity disorder. AB - OBJECTIVE: Recent studies have implicated the involvement of proteins regulating neurotransmitter release in the etiology of attention deficit hyperactivity disorder. On the basis of the role of synapsin III in the modulation of neurotransmitter release, we tested this gene as a candidate contributing to the genetic susceptibility of attention deficit hyperactivity disorder. METHOD: In this study, we genotyped five markers across the gene on 177 small, nuclear families consisting of an attention deficit hyperactivity disorder proband, their parents, and 43 affected siblings. We examined the transmission of the alleles at each one of these sites and the haplotypes of the polymorphisms using the transmission disequilibrium test. RESULT: Our observations did not yield any evidence of biased transmission of the alleles at any polymorphism or haplotype. On the basis of the evidence for synapsins in learning and memory from animal models, we also investigated the relationship of this gene to verbal short-term and working memory as measured by digit span forward and backwards. No evidence was found for an association of this gene to these traits. CONCLUSION: Our findings with this particular sample do not support the synapsin III locus as a major susceptibility locus contributing to attention deficit hyperactivity disorder. PMID- 17413451 TI - MeCP2 gene mutation analysis in autistic boys with developmental regression. AB - Autism and Rett syndrome are both pervasive developmental disorders and share many characteristics in common. One of these features is developmental regression with loss of social, cognitive and language skills after a period of apparently normal development during the first 1-2 years of life, which raises the question of whether there is a common pathway underlying regression in these two disorders. The Rett syndrome gene was identified as MeCP2 gene on Xq28, a powerful transcriptional repressor. To explore its possible role in the etiology of autism and involvement in regression, we searched for MeCP2 gene mutations in a well characterized sample of 31 autistic boys with developmental regression by direct sequencing. One sequence variant in 3' untranslated region was observed. The patient inherited the variant from his unaffected mother, so it may be a rare polymorphism. No coding sequence variant was found in any of the patients tested. We conclude that mutations in the coding sequence of MeCP2 are not a frequent cause of regression in autism. The long 3' untranslated region of MeCP2 is highly conserved across species, suggesting that they are important for the post transcriptional regulation of MeCP2 gene. It may be worthwhile extending the mutation screening, with a larger sample of strictly defined phenotype, to regulatory elements and untranslated regions of this gene, to explore to what degree MeCP2 gene is involved in the etiology of autism and its possible role in the regression of autism. PMID- 17413452 TI - Association study between the cholecystokinin A receptor gene and schizophrenia in the Japanese population. AB - Cholecystokinin A receptor (CCK-AR) has been implicated in the pathophysiology of schizophrenia through its mediation of dopamine-release in the central nervous system. Several studies have observed the association between the CCK-AR gene and schizophrenia. Especially, the association has been repeatedly observed between the 779T/C polymorphism and auditory hallucinations or positive symptoms of schizophrenia. In this study, we investigated the association between the 779T/C polymorphism of the CCK-AR gene and schizophrenia in 290 Japanese patients with schizophrenia and 290 controls. As a result, no significant difference was observed in genotypic distributions or allelic frequencies between the patients and controls, although there was a trend for the association between the C allele of the polymorphism and hallucination (P=0.024) or hallucinatory-paranoid state (P=0.049). In conclusion, the present results may not provide evidence for the association between the CCK-AR gene and schizophrenia in the Japanese population. PMID- 17413453 TI - No association between a common haplotype of the 6 and 10-repeat alleles in intron 8 and the 3'UTR of the DAT1 gene and adult attention deficit hyperactivity disorder. PMID- 17413454 TI - No genetic association between tryptophan hydroxylase 2 gene polymorphisms and Japanese schizophrenia. PMID- 17413455 TI - No association between the serine racemase gene (SRR) and schizophrenia in a German case-control sample. PMID- 17413456 TI - No association between the serine racemase gene (SRR) and bipolar disorder in a German case-control sample. PMID- 17413457 TI - Lumbar facet orientation in spondylolysis: a skeletal study. AB - STUDY DESIGN: Orientation of the lumbar articular facets at the L1-L5 level was measured and analyzed. OBJECTIVE: To characterize the relationship between lumbar facet orientation and isthmic spondylolysis. SUMMARY OF BACKGROUND DATA: Whereas many studies have explored the relationship between facet orientation in the transverse plane and various spinal pathologies, there is insufficient data regarding this relationship and isthmic spondylolysis. METHODS: A 3-dimensional digitizer was used to measure the transverse orientation of the lumbar facet joints at the L1-L5 level in 115 male individuals with bilateral isthmic spondylolysis (at L5) and 120 age and sex-matched normal control subjects from the Hamann-Todd Human Osteological Collection (Cleveland Museum of Natural History, Cleveland, OH). Statistical analysis included paired t tests and analysis of variance. RESULTS: In both isthmic spondylolysis and control groups, considerable shifts were noticed from sagittally oriented articular facets at L1 to frontally oriented facets at L5. The change in orientation was significantly greater (up to 13 degrees at L4) in the isthmic spondylolysis group (right inferior facets). Three of the 4 articular facets of L5 (right and left inferior and right superior) were significantly more frontally oriented in isthmic spondylolysis compared to the control group. A greater tendency of asymmetry in facet orientation was noticed in the isthmic spondylolysis group. CONCLUSION: Individuals with more frontally oriented facets in the lower lumbar vertebrae incorporated with facet tropism are at a greater risk for developing isthmic spondylolysis at L5. PMID- 17413458 TI - Neural space and biomechanical integrity of the developing cervical spine in compression. AB - STUDY DESIGN: A factorial study design was used to examine the biomechanical and neuroprotective integrity of the cervical spine throughout maturation using a postmortem baboon model. OBJECTIVE: To investigate changes with spinal development that affect the neuroprotective ability of the cervical spine in compressive loading. SUMMARY OF BACKGROUND DATA: Child spinal cord injuries claim and debilitate thousands of children in the United States each year. Many of these injuries are diagnostically and mechanistically difficult to classify, treat, and prevent. Biomechanical studies on maturing spinal tissues have identified decreased stiffness and tolerance characteristics for children compared with adults. Unfortunately, while neurologic deficit typically dictates functional outcome, no previous studies have examined the neuroprotective role of the pediatric cervical spine. METHODS: Twenty-two postmortem baboon cervical spines across the developmental age spectrum were tested. Two functional spinal unit segments (Oc-C2, C3-C5, and C6-T1) were instrumented with transducers to measure dynamic changes in the spinal canal. These tissues were compressed to 70% strain dynamically, and the resultant mechanics and spinal canal occlusions were recorded. RESULTS: Classic injury patterns were observed in all of the specimens tested. The compressive mechanics exhibited a significant age relationship (P < 0.0001). Furthermore, while the peak-percent spinal canal occlusion was not age dependent, the percent occlusion just before failure did demonstrate a significant decrease with advancing age (P = 0.0001). CONCLUSIONS: The neuroprotective ability of the cervical spine preceding failure appears to be age dependent, where the young spine can produce greater spinal canal occlusions without failure than its adult counterpart. The overall percent of the spinal canal occluded during a compression injury was not age dependent; however, these data reveal the neuroprotective ability of the child spine to be more sensitive as an injury predictor than the biomechanical fracture data. PMID- 17413459 TI - Stabilizing potential of anterior, posterior, and circumferential fixation for multilevel cervical arthrodesis: an in vitro human cadaveric study of the operative and adjacent segment kinematics. AB - STUDY DESIGN: This is an in vitro biomechanical study. OBJECTIVE: The current investigation was performed to evaluate the stabilizing potential of anterior, posterior, and circumferential cervical fixation on operative and adjacent segment motion following 2 and 3-level reconstructions. SUMMARY OF BACKGROUND DATA: Previous studies reported increases in adjacent level range of motion (ROM) and intradiscal pressure following single-level cervical arthrodesis; however, no studies have compared adjacent level effects following multilevel anterior versus posterior reconstructions. MATERIALS AND METHODS: Ten human cadaveric cervical spines were biomechanically tested using an unconstrained spine simulator under axial rotation, flexion-extension, and lateral bending loading. After intact analysis, all specimens were sequentially instrumented from C3 to C5 with: (1) lateral mass fixation, (2) anterior cervical plate with interbody cages, and (3) combined anterior and posterior fixation. Following biomechanical analysis of 2 level constructs, fixation was extended to C6 and testing repeated. Full ROM was monitored at the operative and adjacent levels, and data normalized to the intact (100%). RESULTS: All reconstructive methods reduced operative level ROM relative to intact specimens under all loading methods (P < 0.05). However, circumferential fixation provided the greatest segmental stability among 2 and 3 level constructs (P < 0.05). Moreover, anterior cervical plate fixation was least efficient at stabilizing operative segments following C3-C6 arthrodesis (P < 0.05). Supradjacent ROM was increased for all treatment groups compared to normal data during flexion-extension testing (P < 0.05). Similar trends were observed under axial rotation and lateral bending loading. At the distal level, flexion extension and axial rotation testing revealed comparable intergroup differences (P < 0.05), while lateral bending loading indicated greater ROM following 2-level circumferential fixation (P < 0.05). CONCLUSIONS: Results from our study revealed greater adjacent level motion following all 3 fixation types. No consistent significant intergroup differences in neighboring segment kinematics were detected among reconstructions. Circumferential fixation provided the greatest level of segmental stability without additional significant increase in adjacent level ROM. PMID- 17413460 TI - Upper extremity palsy following cervical decompression surgery results from a transient spinal cord lesion. AB - STUDY DESIGN: Retrospective analysis. OBJECTIVE: To test the hypothesis that spinal cord lesions cause postoperative upper extremity palsy. SUMMARY OF BACKGROUND DATA: Postoperative paresis, so-called C5 palsy, of the upper extremities is a common complication of cervical surgery. Although there are several hypotheses regarding the etiology of C5 palsy, convincing evidence with a sufficient study population, statistical analysis, and clear radiographic images illustrating the nerve root impediment has not been presented. We hypothesized that the palsy is caused by spinal cord damage following the surgical decompression performed for chronic compressive cervical disorders. METHODS: The study population comprised 857 patients with chronic cervical cord compressive lesions who underwent decompression surgery. Anterior decompression and fusion was performed in 424 cases, laminoplasty in 345 cases, and laminectomy in 88 cases. Neurologic characteristics of patients with postoperative upper extremity palsy were investigated. Relationships between the palsy, and patient sex, age, diagnosis, procedure, area of decompression, and preoperative Japanese Orthopaedic Association score were evaluated with a risk factor analysis. Radiographic examinations were performed for all palsy cases. RESULTS: Postoperative upper extremity palsy occurred in 49 cases (5.7%). The common features of the palsy cases were solely chronic compressive spinal cord disorders and decompression surgery to the cord. There was no difference in the incidence of palsy among the procedures. Cervical segments beyond C5 were often disturbed with frequent multiple segment involvement. There was a tendency for spontaneous improvement of the palsy. Age, decompression area (anterior procedure), and diagnosis (ossification of the posterior longitudinal ligament) are the highest risk factors of the palsy. CONCLUSIONS: The results of the present study support our hypothesis that the etiology of the palsy is a transient disturbance of the spinal cord following a decompression procedure. It appears to be caused by reperfusion after decompression of a chronic compressive lesion of the cervical cord. We recommend that physicians inform patients and surgeons of the potential risk of a spinal cord deficit after cervical decompression surgery. PMID- 17413461 TI - Modified paramedian transpedicular approach and spinal reconstruction for intradural tumors of the cervical and cervicothoracic spine: clinical experience. AB - STUDY DESIGN: Retrospective review of the medical, radiographic, surgical, and postoperative records of patients who underwent resection of multilevel intradural extramedullary spinal cord tumors of the ventral cervical and cervicothoracic spine via a modified paramedian transpedicular approach at the University of California, San Francisco, between 2003 and 2005. OBJECTIVE: To assess the surgical, clinical, and radiographic outcomes of using the modified paramedian transpedicular approach to resect ventral intradural extramedullary spinal cord tumors of the ventral cervical and cervicothoracic spine. SUMMARY OF BACKGROUND DATA: A common theme of skull-base surgery for many years has been to remove the bone rather than retract neural elements. In this report, we demonstrate some possible advantages of taking a "spine-base" approach for resecting intradural extramedullary spinal cord tumors of the ventral cervical and cervicothoracic spinal canal, and present our clinical experience. METHODS: All medical, surgical, and radiologic records were retrospectively reviewed. Clinical outcome was assessed for disability via the Neck Disability Index and for pain via the visual analog scale. RESULTS: Fourteen patients (4 males and 10 females, average age 39.6 years, range 20-62) with intradural extramedullary spinal cord tumors involving multiple levels of the anterior cervical and cervicothoracic spine were identified. All patients presented with pain and/or radiculomyelopathy attributed to a ventral intradural extramedullary spinal cord tumor of the cervical or cervicothoracic spine that was resected via the modified paramedian transpedicular approach with partial dorsal corpectomy and posterior spinal reconstruction. The average follow-up period was 14.6 months (range 5-30). Gross total resection was achieved in all cases, and no patient required additional surgery via an anterior approach for residual tumor. CONCLUSIONS: The modified paramedian transpedicular approach with partial dorsal corpectomy we describe here is a variation of traditional thoracic posterolateral transpedicular extracavitary approaches and offers direct access to lesions of the ventral cervicothoracic spinal canal. This approach avoids the morbidity of anterior transcervical, transoral, or transthoracic procedures, while providing a view of the entire ventral cervicothoracic canal, and can be performed safely and effectively in select patients. PMID- 17413463 TI - Peridural methylprednisolone and wound infiltration with bupivacaine for postoperative pain control after posterior lumbar spine surgery: a randomized double-blinded placebo-controlled trial. AB - STUDY DESIGN: A randomized, double-blind placebo controlled trial in multimodal analgesia for postoperative pain was conducted. OBJECTIVE: To examine whether combination of corticosteroid and bupivacaine administered in patients undergoing posterior lumbosacral spine surgery reduces postoperative morphine consumption, back and leg pain relief, and improves functional disability and general health status. SUMMARY OF BACKGROUND DATA: Patients with lumbar spine surgery had moderate to severe postoperative pain. Administration of corticosteroid or injection of local anesthetic agent has been additive treatment methods for opioid drugs. There is uncertainty as to whether corticosteroid and bupivacaine combination improves outcomes in lumbosacral spine surgery. METHODS: A total of 103 patients who were scheduled to undergo elective posterior lumbar discectomy, decompressive laminectomy with or without instrumented fusion for degenerative spinal diseases, received either methylprednisolone locally applied to the affected nerve roots (and bupivacaine was infiltrated into the wound) or injected placebo. Morphine consumption and pain scores were recorded at 1, 2, 3, 6, 12, 24, and 48 hours after surgery. Oswestry Index and Short Form SF-36 scores were recorded before surgery and at 1 and 3 months later. RESULTS: Demographic data between the 2 groups were comparable. The cumulative morphine dose and postoperative pain was significantly lower in the study group than in the placebo group (P = 0.01 and P = 0.001, respectively). When performing subgroup analyses, the beneficial effects were found in all groups of surgery but could not demonstrated statistically significant difference for all subgroup comparisons. There was no significant difference between the 2 groups with regard to pain on cough, Oswestry Index, and SF-36 scores. No complications were associated with the perioperative use of methylprednisolone or bupivacaine. CONCLUSIONS: Administration of methylprednisolone-bupivacaine provided a favorable effect immediately after posterior lumbosacral spine surgery for discectomy, decompression, and/or spinal fusion without complication. PMID- 17413465 TI - Neck collar, "act-as-usual" or active mobilization for whiplash injury? A randomized parallel-group trial. AB - STUDY DESIGN: Randomized, parallel-group trial. OBJECTIVE: To compare the effect of 3 early intervention strategies following whiplash injury. SUMMARY OF BACKGROUND DATA: Long-lasting pain and disability, known as chronic whiplash associated disorder (WAD), may develop after a forced flexion-extension trauma to the cervical spine. It is unclear whether this, in some cases disabling, condition can be prevented by early intervention. Active interventions have been recommended but have not been compared with information only. METHODS: Participants were recruited from emergency units and general practitioners within 10 days after a whiplash injury and randomized to: 1) immobilization of the cervical spine in a rigid collar followed by active mobilization, 2) advice to "act-as-usual," or 3) an active mobilization program (Mechanical Diagnosis and Therapy). Follow-up was carried out after 3, 6, and 12 months postinjury. Treatment effect was measured in terms of headache and neck pain intensity (0 10), disability, and work capability. RESULTS: A total of 458 participants were included. At the 1-year follow-up, 48% of participants reported considerable neck pain, 53% disability, and 14% were still sick listed at 1 year follow-up. No significant differences were observed between the 3 interventions group. CONCLUSION: Immobilization, "act-as-usual," and mobilization had similar effects regarding prevention of pain, disability, and work capability 1 year after a whiplash injury. PMID- 17413466 TI - Motor neuron involvement in experimental lumbar nerve root compression: a light and electron microscopic study. AB - STUDY DESIGN: The aim of this study is to investigate changes in lumbar motor neurons induced by mechanical nerve root compression using an in vivo model. This study is to investigate the changes of lumbar motor neuron induced by mechanical nerve root compression using in vivo model. OBJECTIVES: The effect of axonal flow disturbance induced by nerve root compression was determined in lumbar motor neuron. SUMMARY OF BACKGROUND DATA: The lumbar motor neuron should not be overlooked when considering the mechanism of weakness, so it is important to understand the morphologic and functional changes that occur in motor neurons of the spinal cord as a result of nerve root compression. However, few studies have looked at changes of neurons within the caused by disturbance of axonal flow, the axon reaction, chromatolysis, and cell death as a result of mechanical compression of the ventral root. METHODS: In mongrel dogs, the seventh lumbar nerve root was compressed for 1 week, or 3 weeks using a clip. Morphologic changes of the motor neurons secondary to the axon reaction were examined by light and electron microscopy. RESULTS: Light and electron microscopy showed central chromatolysis of motor neurons in the lumbar cord from 1 week after the start of compression. After 3 weeks, some neurons undergoing apoptosis were seen in the ventral horn. CONCLUSION: It is important to be aware that, in patients with nerve root compression due to lumbar disc herniation or lumbar canal stenosis, dysfunction is not confined to degeneration at the site of compression but also extends to the motor neurons within the lumbar cord as a result of the axon reaction. Patients with weakness of lower leg should therefore be fully informed of the fact that these symptoms will not resolve immediately after surgery. PMID- 17413467 TI - Proinflammatory cytokines stimulate the expression of nerve growth factor by human intervertebral disc cells. AB - STUDY DESIGN: In vitro studies of the effects of proinflammatory cytokines on the production of nerve growth factor (NGF) by human intervertebral disc (IVD) cells. OBJECTIVE: To determine the constitutive expression and production of NGF and the effect of cytokines on the expression of NGF by human IVD cells. SUMMARY OF THE BACKGROUND DATA: NGF may play a role in the collateral sprouting of sensory axons, neural survival, and regulation of nociceptive sensory neurons. NGF is known to be up-regulated by proinflammatory cytokines. METHODS: The presence of NGF protein was analyzed by immunohistochemistry using human IVD cells obtained from cadaveric human spines with no known disc disease (MRI Thompson grades 2-4). The effects of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF alpha) on NGF production and mRNA expression of NGF by IVD cells were examined. The expression of NGF receptors, trkA and p75, was also assessed immunohistochemically. RESULTS: Cadaveric anulus fibrosus (AF) and nucleus pulposus (NP) cells cultured in vitro in monolayer and in alginate beads positively stained with an anti-NGF antibody. The constitutive production of NGF protein in IVD cells was low (NP) or not detectable (AF). The expression of NGF mRNA was detectable in both cell types. IL-1beta and TNF-alpha up-regulated the NGF mRNA expression and the secretion of NGF protein into the media. TrkA was immunolocalized in AF and NP cells. CONCLUSION: Our results demonstrate, for the first time, that human AF and NP cells constitutively express NGF protein and mRNA, and that the proinflammatory cytokines IL-1beta and TNF-alpha stimulate the production of NGF. The precise role of NGF produced by IVD cells in the generation of discogenic pain or on the metabolism of IVD cells, especially under certain physiologic conditions in which cytokines are up-regulated, needs to be clarified in future experimentation. PMID- 17413468 TI - Atlantoaxial transarticular screw fixation with morselized autograft and without additional internal fixation: technical description and report of 57 cases. AB - STUDY DESIGN: Retrospective clinical and radiologic evaluation of posterior C1-C2 fusion by transarticular screw (TAS) with morselized autograft and without additional internal fixation. OBJECTIVE: Description and assessment of a modified Magerl technique. SUMMARY OF BACKGROUND DATA: The majority of spine surgeons prefer to supplement the posterior TAS with a posterior cable-secured strut graft and a postoperative rigid cervical orthosis. Our hypotheses are that the 2 posterior TASs alone are enough for stabilization and that morselized cancellous grafts have similar clinical result as the structural graft. METHODS: Fifty-seven consecutive patients, including atlantoaxial instability in 52 and atlantoaxial dislocation in 5, were treated by bilateral TAS fixation alone with morselized grafts by the same surgeon. The postoperative external immobilization was abandoned. RESULTS: A total of 114 transarticular screws were placed. Radiographs demonstrated all the screws were placed satisfactorily except two. One screw penetrated into the occipito-atlantal joint, and the other one slightly breached the vertebral artery groove but did not injure vertebral artery. None of these 2 screws was associated with clinical sequelae. There were 2 patients who had postoperative iatrogenic C2-C3 instability on dynamic radiograph, which did not need treatment. These cases had an average follow-up of 47 months (range, 24-76 months). All patients attained solid fusion without screw failure. CONCLUSIONS: Bilateral transarticular screws alone and morselized grafts have high fusion rate in atlantoaxial arthrodesis without instrument failure. TAS fixation could provide stability that is clinically equivalent to the standard screws plus tension band construct as described by Magerl. With anatomic reduction and ideal screw position, additional internal fixation and postoperative collar are not necessary. PMID- 17413469 TI - Surgical strategy for cervical myelopathy due to ossification of the posterior longitudinal ligament: Part 1: Clinical results and limitations of laminoplasty. AB - STUDY DESIGN: Retrospective study of 66 patients who underwent laminoplasty for treatment of cervical myelopathy due to ossification of the posterior longitudinal ligament (OPLL). OBJECTIVES: The present study describes surgical results of laminoplasty for treatment of cervical myelopathy due to OPLL and aims to clarify 1) factors predicting outcome and 2) limitations of laminoplasty. SUMMARY OF BACKGROUND DATA: During the period 1986 and 1996, laminoplasty was the only surgical treatment selected for cervical myelopathy at our institutions. METHODS: We reviewed data obtained in 66 patients who underwent laminoplasty for treatment of cervical myelopathy due to OPLL. Mean duration of follow-up was 10.2 years (range, 5-20 years). Surgical outcomes were assessed using the Japanese Orthopaedic Association (JOA) scoring system for cervical myelopathy. RESULTS: Surgical outcome was significantly poorer in patients with occupying ratio greater than 60%. Multiple regression analysis showed that the most significant predictor of poor outcome after laminoplasty was hill-shaped ossification, followed by lower preoperative JOA score, postoperative change in cervical alignment, and older age at surgery. CONCLUSIONS: Laminoplasty is effective and safe for most patients with occupying ratio of OPLL less than 60% and plateau shaped ossification. However, neurologic outcome of laminoplasty for cervical OPLL was poor or fair in patients with occupying ratio greater than 60% and/or hill-shaped ossification. PMID- 17413470 TI - Surgical strategy for cervical myelopathy due to ossification of the posterior longitudinal ligament: Part 2: Advantages of anterior decompression and fusion over laminoplasty. AB - STUDY DESIGN: Retrospective study of 27 patients who underwent anterior decompression and fusion (ADF) for treatment of cervical myelopathy due to ossification of the posterior longitudinal ligament (OPLL). OBJECTIVES: To compare surgical outcome of ADF with that of laminoplasty. SUMMARY OF BACKGROUND DATA: During the period 1986 and 1996, laminoplasty was the only surgical treatment selected for cervical myelopathy at our institutions. According to surgical results of laminoplasty performed during this period, we have performed either laminoplasty or ADF for patients with OPLL since 1996. METHODS: We reviewed clinical data obtained in 27 patients who underwent ADF between 1996 and 2003. Mean duration of follow-up was 6.0 years (range, 2-10 years). Surgical outcomes were assessed using the Japanese Orthopedic Association (JOA) scoring system for cervical myelopathy. Surgical results of ADF were compared with those of laminoplasty, which was performed in 66 patients during the period 1986 and 1996. RESULTS: ADF yielded a better neurologic outcome at final follow-up than laminoplasty in patients with occupying ratio > or =60%, although graft complications occurred in 15% and additional surgical intervention was required in 26%. Neither occupying ratio of OPLL, sagittal shape of ossification, nor cervical alignment was found to be related to surgical outcome of ADF. CONCLUSIONS: Although ADF is technically demanding and has a higher incidence of surgery-related complications, it is preferable to laminoplasty for patients with occupying ratio of OPLL > or =60%. PMID- 17413471 TI - Long-term results of one-level lumbar arthroplasty: minimum 10-year follow-up of the CHARITE artificial disc in 106 patients. AB - STUDY DESIGN: A retrospective chart and radiographic review. OBJECTIVES: To determine the long-term clinical results, radiographic results, and incidence of complications in a large patient cohort with one-level lumbar total disc replacement (TDR). SUMMARY OF BACKGROUND DATA: Prior authors have described short term, mid-term, and long-term clinical and radiographic results in patients with lumbar TDR with highly variable results. METHODS: From January 1989 to November 1995, 108 patients, with a mean age of 36.4 years, underwent lumbar TDR with the CHARITE Artificial Disc, with 106 available for follow-up. A modified Stauffer Coventry scale was used to determine clinical outcome. Return to work, work level, and the incidence of complications were assessed. Dynamic lateral flexion extension and lateral bending radiographs were performed and segmental range of motion (ROM) was measured using the Cobb method. RESULTS: Mean follow-up time was 13.2 years (range, 10-16.8 years). Of the 106 patients, 87 (82.1%) had either an excellent or good clinical outcome. Of the 96 patients working before surgery, 86 returned to work (89.6%), including 77.8% of patients with hard labor level employment (28 of 36) returning to the same level of work. The mean ROM in flexion-extension was 10.1 degrees , in lateral bending it was 4.4 degrees , and 90.6% of implanted prostheses were still mobile. Eight patients (7.5%) required posterior instrumented fusion. There were 5 cases (4.6%) of postoperative facet arthrosis, 3 cases (2.8%) of subsidence, 3 cases (2.8%) of adjacent-level disease, and 2 cases (1.9%) of core subluxation. CONCLUSIONS: This retrospective study demonstrates the safety and efficacy of the CHARITE Artificial Disc at one level, either L4-L5 or L5-S1, in the long-term. Clinical outcomes and the rate of return to work were excellent overall. The rate of adjacent-level disease requiring surgical intervention was considerably lower (2.8%) compared with reports in the literature for lumbar fusion. As with any surgical procedure, proper indications play a pivotal role in clinical success. PMID- 17413472 TI - Asymmetric expression of melatonin receptor mRNA in bilateral paravertebral muscles in adolescent idiopathic scoliosis. AB - STUDY DESIGN: Comparison of melatonin receptor mRNA expression in bilateral paravertebral muscles in adolescent idiopathic scoliosis (AIS). OBJECTIVES.: To investigate the change of melatonin receptor mRNA expression in bilateral paravertebral muscles in AIS, congenital scoliosis (CS), and control in order to analyze its association to the pathogenesis of AIS. SUMMARY OF BACKGROUND DATA: Muscle imbalance and asymmetry of stretch receptors in the paravertebral muscles of patients with AIS were supposed to have a large role to play in the development and production of the deformity. Melatonin is a focus of studies of the mechanism underlying the development of scoliosis, and there is no research on the expression of melatonin receptors in the paravertebral muscles of patients with AIS. METHODS: Twenty cases with average age of 15.1 +/- 2.2 years and average Cobb angle of 56.2 degrees +/- 16.1 degrees, including 10 cases with Cobb angle >50 degrees and 10 cases with Cobb angle < or =50 degrees, were included in AIS group. The apical vertebrae were from T6 to T11. Twelve cases with an average age of 11.6 +/- 3.2 years and average Cobb angle of 59.2 degrees +/- 33.3 degrees were included in CS group. The apical vertebrae were from T7 to T12. Ten cases without scoliosis were in the control group. The mRNA expression of melatonin receptor subtype MT1 and MT2 was detected by the RT-PCR method. RESULTS: The MT2 mRNA expression on the concave side of the paravertebral muscle was higher than that on the convex side in AIS and CS groups (P < 0.05), but the MT1 mRNA expression showed no significant difference (P > 0.05). In the AIS group, the ratio of MT2 mRNA expression on the concave side compared with the convex side in cases with Cobb angle >50 degrees and cases with Cobb angle < or =50 degrees showed no significant difference (P > 0.05). The MT1 and MT2 mRNA expression showed no significant difference in control group (P > 0.05). CONCLUSION: The melatonin receptor expression in bilateral paravertebral muscles in AIS is asymmetric, which may be a secondary change. The bilateral asymmetry in force exerted on the scoliotic spine may be the cause. PMID- 17413473 TI - Magnetic resonance imaging clarity of the Bryan, Prodisc-C, Prestige LP, and PCM cervical arthroplasty devices. AB - STUDY DESIGN: Prospective, randomized, controlled and double-blinded study on imaging of artificial discs. OBJECTIVE: The purpose of this study is to compare postoperative imaging characteristics of the 4 currently available cervical arthroplasty devices at the level of implantation and at adjacent levels. SUMMARY OF BACKGROUND DATA: Cervical arthroplasty is being performed increasingly frequently for degenerative disc disease and, in most cases, with frank neural compression. Unlike lumbar arthroplasty, performed mainly for axial back pain, decompression of neural elements may need to be confirmed with postoperative imaging after cervical arthroplasty. METHODS: Preoperative and postoperative magnetic resonance imaging scans of 20 patients who had undergone cervical arthroplasty were assessed for imaging quality. Five cases each of the Bryan (Medtronic Sofamor Danek, Memphis, TN), Prodisc-C (Synthes Spine, Paoli, PA), Prestige LP (Medtronic Sofamor Danek), and PCM devices (Cervitech, Rockaway, NJ) were analyzed. Six blinded spinal surgeons scored twice sagittal and axial T2 weighted images using the Jarvik 4-point scale. Statistical analysis was performed comparing quality before surgery and after disc implantation at the operated and adjacent levels and between implant types. RESULTS.: Moderate intraobserver and interobserver reliability was noted. Preoperative images of patients in all implant groups had high-quality images at operative and adjacent levels. The Bryan and Prestige LP devices allowed satisfactory visualization of the canal, exit foramina, cord, and adjacent levels after arthroplasty. Visualization was significantly impaired in all PCM and Prodisc-C cases at the operated level in both the spinal canal and neural foramina. At the adjacent levels, image quality was statistically poorer in the PCM and Prodisc-C than those of Prestige LP or Bryan. CONCLUSIONS.: Postoperative visualization of neural structures and adjacent levels after cervical arthroplasty is variable among current available devices. Devices containing nontitanium metals (cobalt chrome-molybdenum alloys in the PCM and Prodisc-C) prevent accurate postoperative assessment with magnetic resonance imaging at the surgical and adjacent levels. Titanium devices, with or without polyethylene (Bryan disc or Prestige LP), allow for satisfactory monitoring of the adjacent and operated levels. This information is crucial for any surgeon who wishes to assess adequacy of neural decompression and where monitoring of adjacent levels is desired. PMID- 17413474 TI - Disc degeneration in low back pain: a 17-year follow-up study using magnetic resonance imaging. AB - STUDY DESIGN: A prospective, cohort clinical and magnetic resonance imaging (MRI) study of patients with low back pain. OBJECTIVE: To study if lumbar disc degeneration (DD), diagnosed in young patients with low back pain by using MRI will predict chronic pain, disc herniation, or functional disability after a 17 year follow-up. SUMMARY OF BACKGROUND DATA: In 1987, 75 male Finnish conscripts aged 20 years, with low back pain hindering their military service, were studied using MRI at 0.02 T. METHODS: In 2003, 32 patients were reexamined with MRI at 1.0 T. The history of low back illness during the follow-up and current functional outcome were recorded. RESULTS: In 1987, 69% of the 32-patient cohort had DD in one or more lumbar discs. In 2003, all subjects had DD in MRI. The mean number of degenerated discs in each subject increased from 1.1 to 3.0. A total of 76% of discs degenerated in 1987 were herniated in 2003, whereas only 29% of well hydrated discs in 1987 were herniated at the time of follow-up (P = 0.0002). During 17 years of follow-up, 3 patients had undergone spinal surgery. CONCLUSIONS: Early DD in adolescent patients with low back pain predicted the evolution of enhanced DD and herniation in adulthood, but it was not associated with severe low back pain or increased frequency of spinal surgery. PMID- 17413475 TI - The Dynesys lumbar spinal stabilization system: a preliminary report on positional magnetic resonance imaging findings. AB - STUDY DESIGN: We present the positional magnetic resonance imaging findings of a prospective case series of patients undergoing surgery with the Dynesys spinal stabilization device (Zimmer, Inc., Warsaw, IN). OBJECTIVE: To explore the biomechanical impact of the Dynesys device in vivo. SUMMARY OF BACKGROUND DATA: Spinal fusion surgery is widely used for painful degenerative conditions of the lumbar spine that have not responded to conservative measures. It often produces good outcomes but can be associated with adjacent segment hypermobility that may lead to further degeneration and pain. Previous cadaveric biomechanical studies claim that the Dynesys Dynamic Spinal Stabilization System allows some lumbar movement, behaving similar to a normal spine in extension but similar to rigid fixation in flexion. METHODS: Twenty-four patients with dominant low back pain, with or without leg pain, were treated with the Dynesys. All patients underwent positional magnetic resonance imaging before surgery and 9 months after surgery. Measurements were made to assess the differences at the operated level, adjacent level, and whole lumbar spine. RESULTS: There was a statistically significant reduction in flexion-extension range of movement of both the whole lumbar spine by 13.37 degrees (P = 0.002) and at the instrumented segments by 4.08 degrees (P < 0.001) following surgery. There was an insignificant reduction in range of movement at the level above instrumentation (P = 0.807). Mean anterior disc height at the instrumented level reduced by 0.7 mm following insertion of the Dynesys (P < 0.027). Mean posterior disc height reduced by 0.3 mm (P = 0.453). In a neutral posture, the Dynesys had no significant impact on lordosis or inclination of operated or adjacent levels. Contrary to cadaveric study findings, the Dynesys appears to restrict extension more than flexion with respect to a neutral posture. CONCLUSIONS: In vivo, the Dynesys Stabilization System allows movement at the instrumented level, albeit reduced, with no significant increased mobility at the adjacent segments. There was reduction of the anterior disc height without a significant increase of the posterior disc height. PMID- 17413476 TI - Reliability of radiographic parameters in neuromuscular scoliosis. AB - STUDY DESIGN: Retrospective review of radiographic data. OBJECTIVES: This study sought to define interobserver and intraobserver variability to further delineate reliable means by which radiographs of patients with neuromuscular scoliosis can be examined. SUMMARY OF BACKGROUND DATA: Previous studies analyzed the use of Cobb angles in the measurement of idiopathic and congenital scoliosis, but no study until now describes a critical analysis of measurement in evaluating neuromuscular scoliosis. METHODS: Forty-eight patients with neuromuscular scoliosis radiographs were reviewed. These were evaluated for Cobb angle, end vertebrae selection, Ferguson angle, apex of the curve, C7 balance, pelvic obliquity, Risser sign, status of the triradiate cartilage, kyphosis Cobb angle, endplate selection for kyphosis, and kyphotic index. Interclass and intraclass variability was examined with statistical analysis. RESULTS: Cobb angle had an intraobserver variability was 5.7 degrees and the interobserver variability was 14.8 degrees . The intraobserver and interobserver variability for Ferguson angle was 6.8 degrees and 20.6 degrees, respectively. The kyphotic Cobb angle intraobserver variability was found to be 17.4 degrees, and the interobserver variability was 24.01 degrees . CONCLUSIONS: Neuromuscular scoliosis radiographs can be reliably analyzed with the use of Cobb angle. Other forms of analysis, such as Ferguson angle, are not as reliable. Pelvic obliquity should be measured from the horizontal, as other methods are not as reliable. Kyphosis is best evaluated with the use of the kyphotic Cobb angle. Finally, it is felt that a separate anteroposterior pelvis radiograph should be used to assess skeletal maturity, as scoliosis films often truncate the vital anatomy necessary to determine skeletal maturity. PMID- 17413477 TI - Evaluation of the core outcome measure in whiplash. AB - STUDY DESIGN: Reanalysis of data derived from longitudinal cohort studies. OBJECTIVE: To comprehensively evaluate the psychometric properties of a 5-item version of the Core Outcome Measure in people with whiplash. SUMMARY OF BACKGROUND DATA: The 7-item Core Outcome Measure was initially proposed as a brief health outcome measure for use in low back pain. To date, this measure has not been comprehensively assessed in a whiplash population. METHOD.: Data were sourced from 3 separate whiplash cohorts (total 481) encompassing acute, early chronic, and late-chronic whiplash among primary care and insurance populations. Subjects completed a 5-item version of the Core Outcome Measure for whiplash (Core Whiplash Outcome Measure [CWOM]), the Functional Rating Index, Neck Disability Index, SF-36, and perceived recovery questionnaires at baseline and short and long-term follow-up periods. Psychometric evaluation of the CWOM included assessing questionnaire responses, internal consistency, construct validity, and internal and external responsiveness. RESULTS: Internal consistency was excellent at all stages of whiplash (Cronbach alpha = 0.76 in the acute stage and 0.83 in the late-chronic stage). Convergent validity was observed between the CWOM and Functional Rating Index (Pearson r = 0.81), Neck Disability Index (Pearson r = 0.76), and SF-36 physical health summary measure (Pearson r = 0.65). Divergent validity was observed between the CWOM and SF-36 mental health summary measure (Pearson r = -0.45). The internal and external responsiveness of the CWOM was similar to other neck-specific outcome measures. CONCLUSIONS: We recommend the 5-item CWOM as a brief clinical measure for whiplash because it is quick to administer and score, and has excellent measurement properties. The CWOM may need to be supplemented with other questionnaires (e.g., when assessment of psychological or emotional health is required). PMID- 17413478 TI - Video-assisted thoracoscopic surgery in idiopathic scoliosis: evaluation of the learning curve. AB - STUDY DESIGN: Retrospective review of patients with idiopathic scoliosis who underwent same-day or staged anterior and posterior spinal fusion and segmental spinal instrumentation. OBJECTIVE: Evaluation of our learning curve with video assisted thoracoscopic surgery (VATS) with respect to operative time, blood loss, and complications in patients with idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: VATS is a minimally invasive alternative to thoracotomy in the management of idiopathic scoliosis. An increased or steep learning curve has been described in the initial application of this technique. METHODS: We began performing VATS in 1998. We compared our first 25 consecutive VATS patients (Group 2) and subsequent 28 consecutive VATS patients (Group 3) to our previous 16 consecutive patients (Group 1) with a thoracotomy (1991-1998) for idiopathic scoliosis. Training at a sponsored regional course was obtained before performing our first VATS procedure. RESULTS: VATS allowed more disc to be excised in Group 2 (4.5 +/- 1, 5.7 +/- 1, and 4.4 +/- 1 discs in Group 1, Group 2, and Group 3, respectively) and significantly decreased the anterior operative time (215 +/- 33, 260 +/- 56, and 177 +/- 47 minutes) and time per individual disc excision (50 +/- 13, 47 +/- 12, and 41 +/- 12 minutes), while providing comparable correction of the thoracic deformity (67% +/- 12%, 66% +/- 10%, and 70% +/- 13% correction). There was no increase in estimated intraoperative anterior blood loss (228 +/- 213, 183 +/- 136, and 211 +/- 158 mL), estimated blood loss per disc excised (51 +/- 42, 34 +/ 29 and 48 +/- 37 mL), or complications in the VATS groups. Complications were primarily pulmonary and resolved with medical therapy. Postoperative chest tube drainage (855 +/- 397, 462 +/- 249, and 561 +/- 261 mL) and total perioperative anterior blood loss (1083 +/- 507, 647 +/- 309, and 773 +/- 308 mL) were significantly decreased in the VATS groups, but this was attributed to the use of Amicar. CONCLUSIONS: VATS is an effective procedure for anterior spinal fusion in idiopathic scoliosis. The learning curve is short, provided appropriate training is obtained. PMID- 17413480 TI - Re: Dimar JR, Glassman SD, Burkus KJ, et al. Clinical outcomes and fusion success at 2 years of single-level instrumented posterolateral fusions with recombinant human bone morphogenetic protein-2/compression resistant matrix versus iliac crest bone graft. Spine 2006; 31:2534-40. PMID- 17413483 TI - Emergency drills in obstetrics: reducing risk of perinatal death or permanent injury. AB - This article describes the need for mock emergency drills in perinatal emergencies such as shoulder dystocia, maternal hemorrhage, and emergency cesarean section. Effective drills are a patient safety initiative to reduce medical errors and adverse events during the antepartum, intrapartum, and postpartum periods. Successful strategies are identified from other fields of practice to improve patient outcomes. Realistic, institutional specific scenarios for mock emergency drills result in improved team behaviors leading to better outcomes for mothers and infants. PMID- 17413485 TI - A regulatory model on transitioning nurses from education to practice. PMID- 17413486 TI - Filing a state board of nursing complaint against a union of registered nurses during a campaign: was it an expression of union animus or mandatory reporting? PMID- 17413487 TI - Ethics consultation in the emergency department. AB - Clinical ethics teams exist in various forms and have assisted care providers for several decades. Our clinical ethics service at an urban, tertiary, teaching hospital provides ethics consultation to care providers, patients, and their family members. Scenarios prompting an ethics consultation may be complex, often involving social, cultural, and fiscal components. Because patients who receive an ethics consultation often require a lengthy hospital stay, our group searched for unique identifiers in a patient's presentation to facilitate earlier and, potentially, more effective interventions. Of particular interest to our group was the presentation of these patients to our institution from the emergency department (ED). Our group's subjective experience indicated that factors requiring ethics consultation were often present very early during hospitalization. A retrospective medical record review of a convenience sample of 50 records of patients who had received a formal clinical ethics consult within a 14-month timeframe was done. Those patients who were admitted to the hospital via the ED and subsequently received an ethics consultation were identified. The critical issues prompting the ethics consult were then evaluated. Eighteen (35%) of the study patients were originally admitted through the ED. Results showed that the ethical issue(s) that prompted the clinical ethics consult was regularly identifiable in the ED. Our study results indicate that issues prompting ethics consults may potentially be identified as patients present to the ED. Rapid and effective interventions proscribed through institutional policy guidelines could greatly assist nurses and other ED providers in identifying these at-risk patients upon entry of the ED. Such a policy would ultimately benefit both patient and provider. PMID- 17413488 TI - An analysis of phlebitis in a 240-bed hospital. Commemorative Reprint from NITA, Volume 4, No. 4, 1981. PMID- 17413491 TI - Federal, state, and local healthcare systems look to develop, refine, and exercise preparedness plans to protect the public's health from disease, disasters, and bioterrorism. PMID- 17413492 TI - Standard 44: dressings. PMID- 17413493 TI - Liposomal drug delivery. AB - Liposomes are composed of phospholipids, the basic components of human cell walls. Liposome encapsulation improves a medication's bioavailability, which can extend treatment effects and reduce drug dosing. The therapeutic advantages of liposomal drug delivery, such as the ability of long-circulating liposomes to accumulate preferentially at disease sites, including tumors and sites of inflammation, are well recognized. In cases in which a single active has more than one liposome product available, formulation changes leading to differences in pharmacokinetics, toxicity, and clinical efficacy are described. PMID- 17413494 TI - A nursing guide to infusion therapy with abatacept for the treatment of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation of the synovial membranes and progressive joint destruction. Increased understanding of the immunopathology of RA has resulted in the development of new therapies to manage the disease, including several infusion based therapies. Abatacept, a selective costimulation modulator, has been shown to be effective in clinical trials. Abatacept has a mechanism of action that is different from any other biologic RA therapy, and it provides a valuable alternative for RA patients. This article provides practical guidance for nurses to ensure safe administration and to maximize patient outcomes with abatacept. PMID- 17413495 TI - Implementing evidence-based nursing practice in the pediatric intensive care unit. AB - With the widespread use of central venous catheters in children, the incidence of catheter-related bloodstream infections (CR-BSIs) is increasing. Current evidence based practice strategies to decrease CR-BSIs include using maximum barrier techniques during insertion, practicing good hand hygiene, performing skin antisepsis with 2% chlorhexidine, using a chlorhexidine-impregnated patch (CIP) covered by a semipermeable polyurethane dressing, and promptly removing catheters when no longer needed. Implementation of evidence-based practice bundles, along with monthly monitoring of infection surveillance, has resulted in significant decreases in the average rates of CR-BSIs per 1,000 catheter days in many pediatric intensive care units. PMID- 17413496 TI - Current therapies for multiple myeloma. AB - Multiple myeloma is an incurable B-cell malignancy of plasma cells but also a highly treatable disease. This article is an overview of multiple myeloma and its current therapies, with emphasis on bortezomib and lenalidomide. Important nursing considerations related to the management of common adverse events are also described. PMID- 17413501 TI - Roundup: perspectives on a perfect storm. AB - This article offers perspectives on the nursing shortage and how to deal with them from a variety of healthcare leaders. PMID- 17413502 TI - The perfect storm: managed care, aging adults, and a nursing shortage. AB - The many issues managed care poses for providers and health networks are crystallized in the moral problems occasioned by its shifting of the financial risks of care from the insurer to the provider. Yet, of 257 data-based studies of nursing care quality identified, 135 investigated a process-outcome link but only 17 met study inclusion criteria. The literature provides evidence that the quality of nursing care processes affects health-related patient outcomes during and after hospitalization. Thus, California became the first state to mandate nurse staffing ratios on all units in 1997. All the while labor shortages, introduction of new technology, and the increased acuity of the inpatient population have contributed to the rising costs of hospital-based care. Another notable contribution to the financial and clinical pressures hospitals feel is the overall increased consumer demand for services: The aging of America's population portends an even greater demand for healthcare services. This situation will only grow worse as the demand generated by the growing gerontocracy increasingly outstrips an affordable supply of safe care. Little time and few resources are available for what we might all agree would be the desired level of comprehensive and coordinated care. What this means for patients is that they experience episodic acute care that is delivered in "spurts." No matter what actions are taken - or more likely not taken - to reform the Medicare program, the future holds shrinking reimbursement for hospitals, with a concurrent demand for investments in response to costly regulatory requirements, technologies, and other capital expenditures to accommodate the increasing demand for services generated by the growing numbers of politically active seniors who insist on the entitlements promised to them - and the quality of service they believe is their right. This is the Perfect Storm.... PMID- 17413503 TI - Key forecasts shaping nursing's perfect storm. AB - Perfect storms abound in nursing and healthcare. How we plan for them and how we forecast effectively which ones will have tremendous impact on how we lead the profession is a challenge to anyone who is or will be a leader. This article focuses on key forecasts that contribute to creating perfect storms of the future. The "perfect storm" is a term found in multiple disciplines. The phrase denotes the condition that exists when events occur simultaneously with the result that this confluence has a greater impact than what could have resulted from a chance combination. Although perfect storms are rare, they have enormous impact when they occur, and if an alteration in any of the events occurs, the overall impact is lessened. PMID- 17413504 TI - Confronting the perfect storm: a challenge to nurse leaders. AB - As leaders confront the issues and impending problems of today and the future, their number and complexity may overwhelm some. The individuals and organizations who will energize themselves and others to meet the challenges of healthcare are those who maintain quality patient care as their central priority, have a bias for action, and collaborate with other stakeholders. In this article, the immediate past president of the American Organization of Nurse Executives describes how some organizations are doing just that. PMID- 17413505 TI - Evidence and root causes of an inadequate pipeline for nursing faculty. AB - The North Carolina Center for Nursing (NCCN) examined the projected supply of nursing faculty in the state of North Carolina. Coupled with a longitudinal educational mobility study of the state's registered nurses, the forecast shows that the growing faculty shortage is real and that its root cause is a growing shortfall in the pipeline of RNs prepared educationally to pursue graduate education and assume faculty roles. PMID- 17413506 TI - HIT plants SEEDS in healthcare education. AB - By incorporating a clinical information system in the education curriculum as a teaching platform, the University of Kansas School of Nursing teaches nurses and other health professional students how to assess, plan, document and manage care in an electronic medium that develops healthcare informatics competencies. The outcomes of this integrated technology curriculum brings hope for transforming health professional education for 21st century practice and graduating a workforce with the leadership and competencies for improving quality and safety in patient care. It results in IT savvy healthcare providers who will cross the quality chasm. PMID- 17413507 TI - Burdensome situations in everyday nursing: an explorative qualitative action research on a medical ward. AB - Everyday nursing care is under marked constraint in the current healthcare environment due to decreasing time resources and competing financial demands. Burden along with stress and burnout is increasing. Stress not only does occur on the individual and organizational level, as has been studied, but also is a significant factor at the team level, particularly with regard to interactions at this level. This project aimed at eliciting nurses' view on burden; identifying areas for intervention on team level; and evaluating the interventions implemented at this level. Focus groups were conducted with 2 nursing teams (n = 36). Data were analyzed with Atlas Ti, Version 4.2. On the basis of the results of the focus groups, interventions were developed and implemented observing the tenets of problem-based learning and evidence-based nursing. Verbal feedback was used to evaluate the impact of the interventions on the nursing team. Fourteen themes were identified for each ward as areas for improvement. Oral evaluation of the group-specific interventions revealed a benefit for the nursing team as a whole. Findings indicate that team actions played an important role when addressing stressful nursing situations. To improve stressful nursing situations, team action needs to be considered. PMID- 17413508 TI - Sleepy nurses: are we willing to accept the challenge today? AB - With a shortage of supply of nurses and the increasing demand for nursing care, hospitals require or allow nurses to work extended shifts (in excess of 12 hours) and many shifts per week (up to and in excess of 60 hours per week). The result of these excessive hours of work is that many nurses care for patients while suffering from sleep deprivation. Sleep deprivation has been shown to negatively impact judgment and performance resulting in errors and accidents. Sleep deprivation also negatively affects the health of individuals. Sleep deprivation in nurses is a significant issue that requires more attention. PMID- 17413509 TI - Assessing the quality of nursing work life. AB - Traditionally, nursing has measured job satisfaction by focusing on employees' likes and dislikes. However, job satisfaction is an unsatisfactory construct to assess either the jobs themselves or employees' feelings about work sinceas much as 30% of the variance explained in job satisfaction surveys is a function of personality, something employers can do little to change. Based on socio technical systems theory, quality of nursing work life (QNWL) assessments focus on identifying opportunities for nurses to improve their work and work environment while achieving the organization's goals. Moreover, some evidence suggests that improvements in work life are needed to improve productivity. Therefore, assessing QNWL reveals areas for improvement where the needs of both the employees and the organization converge. The purpose of this article was to assess the QNWL of staff nurses using Brooks' Quality of Nursing Work Life Survey. PMID- 17413510 TI - Beyond caring. AB - "Can caring be taught?" is a question that spurs an emotional debate among nurses and leads to further questions: Is nursing science overshadowed by caring in nursing practice? Is the scientific basis of the nursing profession overlooked as a result of the emphasis of caring in nursing? Is it a logical fallacy to assume a correlation between nursing science and caring? Can nursing science or caring alone provide the basis of nursing practice? Further investigation is necessary into the depth of caring and its impact on nursing science and practice. PMID- 17413511 TI - A conceptual model for growing evidence-based practice. AB - Nursing administration at a small medical center is developing and implementing an evidence-based practice (EBP) model of care to support a culture of quality care, clinical excellence, cost-effectiveness, critical thinking, empowerment of staff, and professional growth. The purpose of this article is to describe a conceptual model for EBP that addresses how to overcome barriers to implementation. Clinician expertise and values, experience, patient preference and expectation, and caring become grounded in a practice environment that must strive to become rooted in clinical research to evolve into a practice that is evidence-based. Education helps to nourish EBP, but leadership, enthusiasm, mentorship, clinical inquiry, and reflective practice make EBP thrive. The EBP ambassadors branch out to each department to grow journal clubs, EBP Web pages, EBP projects, research utilization projects, and staff-led practice reviews. The fruits are quality patient care and outcomes, clinical excellence, cost effectiveness, critical thinking, empowerment of staff, and professional growth. PMID- 17413512 TI - The coaching process: an effective tool for professional development. AB - A model for coaching in nursing is described. Criteria for selecting a coach are discussed. Competencies for a coach are recommended. In addition, guidelines for caching sessions are provided as well as an example of an action plan outline to help the coachee identify areas of desired growth and options for developing these areas. PMID- 17413513 TI - Understanding delegation is a critical competency for nurses in the new millennium. PMID- 17413514 TI - Elbow dislocations associated with ipsilateral radial shaft fractures: a case report and review of the literature. PMID- 17413515 TI - Laparoscopic splenectomy for delayed splenic rupture after embolization. PMID- 17413516 TI - Successful management of pulmonary artery perforation after chest tube insertion. PMID- 17413517 TI - Grossly shortened and cross-united forearm fracture treated by gradual distraction using a simple external fixator. PMID- 17413518 TI - Low-molecular-weight heparin for prevention of thrombosis: inverted role. AB - BACKGROUND: Heparin induced thrombosis (HIT) after low-molecular-weight (LMWH) administration for thrombosis prevention is a limb and life threatening condition. METHODS: Two previously healthy individuals, with nonviable forearm amputation and knee osteoarthritis, suffered from multiple arterial thromboses (>2 sites each) after postoperative administration of LMWH. RESULTS: Both patients suffered from thrombocytopenia on the first postoperative days (4-7) and had signs of arterial occlusions on the 9th and 10th postoperative days. The patient with the initially successful forearm replantation was finally amputated below the elbow, whereas the lower extremity of the patient with total knee arthroplasty and contralateral femoral artery occlusion was salvaged with foot fingers amputation only. CONCLUSIONS: Early recognition of the syndrome by monitoring daily platelet counts during heparin therapy is essential. In case of postheparin (LMWH or unfractionated) platelet decrease, heparin administration should be ceased after emergency testing for heparin induced antibodies. When HIT syndrome is confirmed to have occurred, heparin/LMWH must be replaced with thrombin-specific inhibitors. PMID- 17413519 TI - Deep vein thrombosis and fatal paradoxical embolism after delayed reduction of a traumatic hip dislocation. PMID- 17413520 TI - Total inflow occlusion to repair a penetrating cardiac injury: case report. PMID- 17413521 TI - Subcutaneous unilateral testicle implantation following contralateral orchiectomy and radical debridement for Fournier's gangrene: 24-year follow-up. PMID- 17413522 TI - Use of FloSeal matrix hemostatic agent in partial splenectomy after penetrating trauma. PMID- 17413523 TI - Bilateral exercise induced exertional compartment syndrome resulting in acute compartment loss: a case report. PMID- 17413524 TI - Nonoperative management of post-traumatic pulmonary pseudocyst after severe thoracic trauma and hemorrhage by coagulation management, kinetic therapy, and control of secondary infection: a case report. PMID- 17413525 TI - Ligation of the subclavian artery after blunt trauma presenting as massive hemothorax. PMID- 17413526 TI - Acute myocardial infarction caused by occult coronary intimal dissection after a heel stomp: a case report. PMID- 17413527 TI - Tracheal rupture after air bag deployment. PMID- 17413528 TI - Off-pump repair of LAD transection and LV rupture for cardiac stab wound. PMID- 17413529 TI - Colorectal and vaginal injuries in personal watercraft passengers. PMID- 17413530 TI - Endoscopic third ventriculostomy in pediatric patients: the Canadian experience. AB - OBJECTIVE: Reports from relatively small series of pediatric patients predominantly from single centers have hampered accurate analysis of outcome from endoscopic third ventriculostomy. We combined patients from nine pediatric neurosurgery centers across Canada to obtain a better estimate of outcome and identify factors affecting success of the procedure. METHODS: Databases were recoded for uniformity. Failure of the procedure was defined as any subsequent operation or death resulting from hydrocephalus. Time to failure was analyzed by Kaplan-Meier estimate and Cox proportional hazard analysis. RESULTS: During a 15 year period (1989-2004), 368 patients underwent the procedure. The average age was 6.5 years, and 57% were male. Aqueduct stenosis and tumors were the most common etiology, comprising 34 and 29%, respectively. Twenty-two percent of the patients had been previously shunted. The 1- and 5-year success rates were 65 and 52%, respectively. Factors included in the Cox model were age, sex, etiology of hydrocephalus, previous surgery, center volume, and surgeon volume. By multivariate analysis, only age had a significant effect on outcome, with younger patients failing at higher rates, particularly neonates and infants. CONCLUSION: Based on data from multiple Canadian centers, age seems to be the primary determinant of outcome in endoscopic third ventriculostomy in pediatric patients. Failure rates are particularly high in neonates and young infants; thus, the role of this procedure in this age group should be carefully considered. PMID- 17413534 TI - Vaginal douching and development of bacterial vaginosis among women with normal and abnormal vaginal microflora. AB - OBJECTIVES: To evaluate the relationship between douching and bacterial vaginosis (BV) among women with and without prior abnormal vaginal flora. STUDY DESIGNS: 1193 women had vaginal swabs obtained for Gram stain for BV, culture for vaginal microflora, and DNA amplification for Neisseria gonorrhoeae and Chlamydia trachomatis at baseline and 6, 12, 24, and 36 months. Self-reported douching was queried every 6 months. RESULTS: In cross-sectional analyses, douching at least once per month was associated with BV among women who had BV at the immediately preceding visit but not among women with normal or intermediate flora. In prospective analyses, douching increased the risk of acquisition for BV among women with intermediate flora (adj. HR 1.5, 95% CI 1.1-2.4) but not among women with normal flora. CONCLUSIONS: Douching appeared to be associated with BV among women with already imbalanced flora but not among women with normal flora. PMID- 17413535 TI - Incidence of neonatal herpes simplex virus infections in a managed-care population. AB - OBJECTIVE: To determine the incidence of possible neonatal herpes simplex virus (HSV) infections, HSV infection status of women with infected infants, and use of measures to reduce risk of HSV transmission to the neonate in a large US managed care population. STUDY DESIGN: Retrospective analysis of administrative claims from the Integrated Health Care Information Services National Managed Care Benchmark database. RESULTS: Of 233,487 infants born to 252,474 mothers from January 1997 to June 2002, the numbers assigned an ICD-9 code reflecting possible neonatal HSV infection VC), and, thus far, 21 (27%) are able to GPB. Fifteen could GPB sufficiently to delay onset of daytime ventilator use and, later, to require 1.9 fewer ventilator assisted breaths per minute. For the 47 patients with multiple data points, as VC deteriorated from 1080 +/- 870 to 1001 +/- 785 ml, MIC increased from 1592 +/- 887 to 1838 +/- 774 ml. For 21 patients, GPmaxSBC significantly exceeded VC (824 +/- 584 vs. 244 +/- 151 ml, respectively, P < 0.001). The ability to increase lung volume by air stacking (MIC) was better retained than was the ability to increase lung volume by GPB (GPmaxSBC). Air stacking also permitted assisted CPF to exceed unassisted CPF: 289 +/- 91 and 164 +/- 76 liters/m, respectively (P < 0.001). CONCLUSIONS: GPB and air stacking can increase lung volumes and, thereby, cough flows. GPB also can be used in many cases to delay and decrease daytime ventilator use. PMID- 17413543 TI - Air doping: an expose on "frog" insufflation in competitive sports. PMID- 17413544 TI - High-resolution magnetoencephalographic functional mapping of the cortical network mediating intentional movement. AB - Magnetoencephalography (MEG) is a sensitive technique that can detect and map cortical electrophysiologic activations with high spatial (mm) and temporal (msecs) resolutions. We used 148-channel whole-head MEG to record the activation sequence for the somatosensory and motor cortical network during cued hand movements in a healthy 39-yr-old subject. The complex sequence and topography of cortical activations were superimposed onto the subject's brain magnetic resonance images. Frontal premotor and supplementary motor and cingulate areas activated well before the primary motor area and again repetitively from 200 msecs onward with activations alternating repeatedly between frontal and parietal areas. The network's very close functional integration of supplementary motor areas suggests how brain injury that is localized to these regions, but not to the primary motor area itself, can disrupt integrity of movement, and why preservation of functional integrity of some areas traditionally viewed as extramotor may be necessary for recovery from neurologic disability. PMID- 17413545 TI - Introduction to musculoskeletal diagnostic ultrasound: examination of the upper limb. AB - With recent advances in computer technology and equipment miniaturization, the clinical application of diagnostic ultrasonography (U/S) has spread across various medical specialties. Diagnostic U/S is attractive in terms of its noninvasiveness, lack of radiation, readiness of use, cost-effectiveness, and its ability to make dynamic examinations possible. Dynamic imaging deserves special emphasis because it is useful in differentiating full-thickness from partial thickness tendon tears, muscle tears, and tendon and nerve subluxations or dislocations. It is also a quick and easy avenue for side-to-side comparisons. When appropriately used, diagnostic U/S can be considered as an extension of one's physical examination. However, there are limitations of U/S, which will be discussed in this review article. This is part 1 of two articles; this first part will focus on the ultrasound examination of the upper extremity, using selected examples relevant to musculoskeletal medicine. Part 2 will cover common pathologies of the lower extremity. PMID- 17413546 TI - Synovitis of the sternoclavicular joint: the role of ultrasound. PMID- 17413547 TI - Quantification of pulsatility as a function of vascular input impedance: an in vitro study. AB - The physiological benefits of pulsatility generated by ventricular assist device (VAD) support continue to be heavily debated as application of VAD support has been expanded to include destination and recovery therapies. In this study, the relationship between input impedance (Zart) and vascular pulsatility during continuous flow (CF) or pulsatile flow (PF) VAD support was investigated. Hemodynamic waveforms were recorded at baseline failure and with 50%, 75%, and 100% CF or PF VAD support for nine different Zart test conditions (combination of three different resistance and compliance settings) in a mock circulatory system simulating left ventricular failure. High-fidelity hemodynamic pressure and flow waveforms were recorded to calculate mean arterial pressure (MAP), Zart, energy equivalent pressure (EEP), and surplus hemodynamic energy (SHE) as metrics for quantifying vascular pulsatility. MAP and EEP were elevated with increasing resistance whereas SHE was reduced with increasing compliance. Vascular pulsatility was restored with increasing PF VAD support, but diminished by up to 90% with increasing CF VAD support. The nonpulsatile energy component (MAP) of the pressure waveform is dependent on resistance whereas the pulsatile energy component (SHE) is dependent on compliance. The impact of Zart and vascular pulsatility on patient recovery with VAD support warrants further investigation. PMID- 17413548 TI - Development and validation of a computational fluid dynamics methodology for simulation of pulsatile left ventricular assist devices. AB - An unsteady computational fluid dynamic methodology was developed so that design analyses could be undertaken for devices such as the 50cc Penn State positive displacement left ventricular assist device (LVAD). The piston motion observed in vitro was modeled, yielding the physiologic flow waveform observed during pulsatile experiments. Valve closure was modeled numerically by locally increasing fluid viscosity during the closed phase. Computational geometry contained Bjork-Shiley Monostrut mechanical heart valves in mitral and aortic positions. Cases for computational analysis included LVAD operation under steady flow and pulsatile-flow conditions. Computations were validated by comparing simulation results with previously obtained in vitro particle image velocimetry (PIV) measurements. The steady portion of the analysis studied effects of mitral valve orientation, comparing the computational results with in vitro data obtained from mock circulatory loop experiments. The velocity field showed good qualitative agreement with the in vitro PIV data. The pulsatile flow simulations modeled the unsteady flow phenomena associated with a positive-displacement LVAD operating through several beat cycles. Flow velocity gradients allowed computation of the scalar wall strain rate, an important factor for determining hemodynamics of the device. Velocity magnitude contours compared well with PIV data throughout the cycle. Computational wall shear rates over the pulsatile cycle were found to be in the same range as wall shear rates observed in vitro. PMID- 17413549 TI - Home discharge experience with the Thoratec TLC-II portable driver. AB - With the growing success and expanded use of ventricular assist devices, home discharge with independent ambulation and self-care are now important issues. We describe our initial home discharge experience with the Thoratec TLC-II portable drive. Patients discharged home were required to have five outpatient excursions (three monitored) before discharge and were seen weekly after discharge. Between August 2000 and December 2004, 14 patients (average age, 57 years) were placed on the TLC-II portable driver. One patient on the TLC-II portable driver had left ventricular assist device removal after 50 days of support but before discharge. Thirteen patients were discharged from the hospital with average time at home of 62 days (range, 16 to 243 days). After discharge, the TLC-II portable driver was maintained in the auto mode at average settings of 78 beat rate, 5.1 L/min flow rate, 204 mm Hg ejection pressure, and -8.4 mm Hg fill vacuum. A total of 5852 alarms (average, 6.7 per day) were recorded with 2373 battery reminders (41%), 1922 occlusion alarms during sleep (33%), and 1461 no-fill signals (25%). There were no readmissions for device malfunction, emergency battery utilization or back-up unit use. These results demonstrate that the Thoratec TLC-II portable driver is safe, reliable, and can be effectively managed at home. PMID- 17413550 TI - Feasibility study of a direct endo-aortic clamp balloon. AB - We have developed a new end-aortic clamp balloon catheter intended to be inserted directly into, thereby occluding, the ascending aorta. We examined the performance of this catheter in a canine model. We evaluated the extent of migration tolerance of the catheter under cardiopulmonary bypass perfusion in 12 mongrel dogs, weighing 20 kg, under general anesthesia. After institution of cardiopulmonary bypass, this catheter was inserted into the ascending aorta, and the balloon was inflated to occlude the ascending aorta. After the canine heart was arrested following the administration of cardioplegic solution, balloon migration was examined over a period of 3 hours, with hourly increases in perfusion pressure from 50 mm Hg to 80 mm Hg and finally to 100 mm Hg. After the migration test, ascending aortic wall sections, where the balloon was inflated, were examined microscopically. At internal balloon pressure of 300 to 400 mm Hg, migration occurred at perfusion pressure of > or =90 to 100 mm Hg. No histological differences were observed with use of the balloon catheter, compared with an extra-aortic clamp forceps. Based on these results, this device is safe, feasible, and can adequately occlude the ascending aorta during cardiopulmonary bypass. We conclude that this device is effective in patients weighing 20 kg. PMID- 17413551 TI - Minimal sensor count approach to fuzzy logic rotary blood pump flow control. AB - A rotary blood pump fuzzy logic flow controller without flow sensors was developed and tested in vitro. The controller, implemented in LabView, was set to maintain a flow set point in the presence of external pressure disturbances. Flow was estimated as a function of measured pump's delta P and speed, using a steady state, nonlinear approximation. The fuzzy controller used the pump's flow estimate and delta P as feedback variables. The defuzzified control output manipulated the pump speed. Membership functions included flow error, delta P, and pump speed. Experimental runs in a mock loop (water/glycerin 3.5 cPs, 37 degrees C), using the estimated flow, were compared with those using a Transonic flow meter for nine conditions of flow and delta P (4 to 6 L/min, 150 to 350 mm Hg). Pressure disturbances generated by a servo pinch valve ranged from +/-23 to +/-47 mm Hg. Results indicated that the fuzzy controller ably regulated the flow set point to within +/-10% of the baseline even under large swings in pressure. There was no difference in controller performance between the ultrasonic flow measurement and the estimated flow calculation scenarios. These tests demonstrated that the fuzzy controller is capable of rejecting disturbances and regulating flow to acceptable limits while using a flow estimate. PMID- 17413552 TI - Comparison of LionHeart LVAD-implanted calves based on renal infarctions. AB - Calves implanted with the LionHeart left ventricular assist device during the Food and Drug Administration Investigational Device Exemption (IDE) trial had fewer renal infarctions than calves implanted after this trial (post-IDE). Since a decrease in renal infarctions indicates a favorable outcome, we compared nine IDE with 16 post-IDE implantations. Renal infarct severity was graded (0 to 4) on the basis of necropsy reports, histopathology reports, and photographs. Implants were compared for duration of support, anticoagulation, device flow, percent fill, device rate, free plasma hemoglobin, fibrinogen, platelet count, nonsteroidal anti-inflammatory drug use, inlet cannula shape, and outlet graft-to graft anastomosis technique. IDE calves had lower renal pathology scores (1.8 vs. 2.8, p = 0.03), mean device percent fill (88.5% vs. 94.0%, p = 1 x 10), and median free plasma hemoglobin (2.6 vs. 4.4 mg/dL, p = 0.01). IDE animals also had a higher mean device rate (108 vs. 98 bpm, p = 0.04), received low-molecular weight dextran in the early postoperative period, began oral warfarin earlier, had straighter inlet cannulae, and had factory-sewn outlet graft-to-graft anastomoses. Duration of support did not predict severity of renal infarction, and there were no differences in device flow, international normalized ratio, platelet count, or nonsteroidal anti-inflammatory drug use. These results could assist future implantations. PMID- 17413553 TI - Left ventricular assist device support induces acute changes in myocardial electrolytes in heart failure. AB - The regulation of myocardial electrolyte concentrations is critical to proper cardiac function. Myocardial ischemia is associated with deranged ion transport. Left ventricular assist device (LVAD) therapy improves myocyte bioenergetics in chronic heart failure (CHF), which may manifest as electrolyte alterations; however, rapid electrolyte shifts may place critically ill patients at risk for arrhythmias upon initiation of LVAD support. We examine the effect of incremental increases in LVAD support on acute changes in myocardial arteriovenous electrolytes in CHF. CHF was induced in sheep via coronary microembolization. Four months later, sheep underwent acute LVAD implantation. LVAD support was incrementally increased (0%, 25%, 50%, 75% support). Paired arterial and coronary sinus blood samples were obtained at each increment and analyzed for K+, Ca2+, and Na+ concentrations. Arteriovenous electrolyte concentrations (mmol/l) were inverted in CHF before LVAD support: K+ (-0.08), Ca2+ (-0.04), and Na+ (0.04). These imbalances were corrected within 20 minutes and with as little as 25% LVAD support: K+ (0.06), Ca2+ (0.012), and Na+ (-0.80). The arteriovenous differences further widened as LVAD support was increased. In conclusion, LVAD support in CHF induces acute alterations in myocardial electrolytes. Rapid shifts myocardial arteriovenous electrolyte balances during LVAD support may in part explain the incidence of post-LVAD arrhythmias observed clinically in humans. PMID- 17413554 TI - An in vitro method for assessing biomaterial-associated platelet activation. AB - The development of a nonthrombogenic artificial surface for use with indwelling sensors or catheters remains an elusive goal despite decades of ongoing research. In vivo studies are both labor intensive and costly, and are therefore an inefficient way to rapidly screen possible surface materials. The following in vitro model used glass, polyvinyl chloride (PVC), and polypropylene test tubes incubated with 111In-labeled rabbit platelets and illustrated that, despite equivalent platelet count and function, platelet adhesion was greatest on glass (n = 13), with PVC (n = 17) at 67 +/- 8% and polypropylene (n = 13) at 43 +/- 5% when compared with glass. Extrapolating this method by coating test tubes with new, nonthrombogenic materials is a quick and reliable way to screen material before embarking upon more lengthy in vivo animal studies. PMID- 17413555 TI - Carbon dioxide clearance in rabbits during expiratory phase intratracheal pulmonary ventilation. AB - The purpose of this study was to compare the efficacy of CO2 removal during conventional mechanical ventilation (CMV) with and without expiratory phase intratracheal pulmonary ventilation (expiratory ITPV or Exp-ITPV); and to compare CO2 clearance during Exp-ITPV, in pressure-controlled ventilation (PCV) and in volume-controlled ventilation (VCV) modes. Seven anesthetized rabbits were tracheotomized and intubated using a 4 mm endotracheal tube. Venous and arterial lines were established. The rabbits were paralyzed, mechanically ventilated, and ventilation parameters were adjusted to achieve baseline arterial hypercapnia. Animals were then ventilated during 30-minute trials of CMV and Exp-ITPV, in both PCV and VCV modes. A custom-built, microprocessor-controlled solenoid valve was used to limit ITPV gas flow to the expiratory phase. Proximal and carinal airway pressures and hemodynamic variables were continuously recorded, and arterial blood gases were analyzed at the end of each trial. Exp-ITPV, as compared with CMV, reduced arterial PCO2 by 12% and 21% in PCV and VCV modes, respectively (p < 0.02 and p < 0.001; one-sided paired t test), without significant changes in other cardiorespiratory variables. In conclusion, Exp-ITPV is more effective than CMV in clearing CO2 through a small endotracheal tube. Exp-ITPV is also more effective in VCV mode than PCV mode. PMID- 17413556 TI - Bridge to lung transplantation with the extracorporeal membrane ventilator Novalung in the veno-venous mode: the initial Hannover experience. AB - Conventional extracorporeal membrane oxygenation and mechanical ventilation have both been identified as significant risk factors for post-lung transplant mortality when applied as a bridge to lung transplantation. We have previously described the successful use of the extracorporeal membrane ventilator Novalung as a bridge to lung transplantation in patients with severe hypercapnia and respiratory acidosis. In this setting the Novalung was connected in the arterio venous mode without support of a mechanical blood pump. However, in patients with predominant hypoxemia, this pumpless mode does not achieve sufficient blood oxygenation due to limited blood flow. Thus, such patients require pump-driven support. Here we describe our initial experience with the Novalung extracorporeal membrane ventilator, which was originally designed for pumpless carbon dioxide removal, as a bridge to lung transplantation in patients with ventilator refractory hypoxemia in the veno-venous pump-driven mode. PMID- 17413557 TI - Development of "plug and play" TransApical to aorta VAD. AB - Our TransApical to Aorta pump, a simple and minimally invasive left ventricular (LV) assist device, has a flexible, thin-wall conduit connected by six struts to a motor with ball bearings and a turbine extending into the blood path. Pulsatile flow is inherent in the design as the native heart contraction preloads the turbine. In six healthy sheep, the LV apex was exposed by a fifth intercostal left thoracotomy. The pump was inserted from the cardiac apex through the LV cavity into the ascending aorta. Aortic and LV pressure waveforms, pump flow, motor current, and pressure were directly measured. All six cannula pumps were smoothly advanced on the first attempt. Pump implantation was <15 minutes (13.6 +/- 1.8 minutes). Blood flow was 2.8 l/min to 4.4 l/min against 86 +/- 8.9 mm Hg mean arterial blood pressure at maximum flow. LV systemic pressure decreased significantly from 102.5 +/- 5.55 mm Hg to 58.8 +/- 15.5 mm Hg at the fourth hour of pumping (p = 0.042), and diastolic LV pressure decreased from 8.4 +/- 3.7 to 6.1 +/- 2.3 mm Hg (p > 0.05). The pump operated with a current of 0.4 to 0.7 amps and rotation speed of 28,000 to 33,000 rpm. Plasma free hemoglobin was 4 +/- 1.41 mg/dl (range, 2 to 5 mg/dl) at termination. No thrombosis was observed at necropsy.A left ventricular assist device using the transapical to aorta approach is quick, reliable, minimally invasive, and achieves significant LV unloading with minimal blood trauma. PMID- 17413558 TI - Tissue engineering of an implantable bioartificial hemofilter. AB - The first step in the tissue engineering of an implantable bioartificial kidney is the development of an implant that produces ultrafiltrate to replace glomerular function. A fabricated device containing synthetic hollow hemofiltration fibers was placed around the femoral vascular pedicle in rats, which initiated new tissue formation with a mature and durable neocapillary bed. The transudate fluid produced by this newly formed capillary bed accumulated through the hollow fibers into a subcutaneous port to allow evaluation of the fluid. In its first phase, this study evaluated various hollow fibers and tissue induction processes by the measurement of fluid volume, urea nitrogen, and total protein continuously for 6 weeks. New tissues formed within the implants surrounding the fibers, and the vascular density, vessel sizes, and percent cross sectional vascular area were assessed by means of histomorphometric analysis after 6 weeks. The volume of fluid formation correlated with both vascular density and fiber membrane surface area. The implant fluid-to-serum ratios demonstrated a permselective filtrate. In a second phase, platelet-derived growth factor and vascular endothelial growth factor versus carrier alone were infused directly into the implants for the first 4 weeks in vivo through osmotic pumps and followed up to 9 weeks. Cumulative implant fluid volumes were significantly greater in the growth factor-treated group than in control animals and were associated with greater numbers of small-caliber blood vessels. These results provide the initial proof of concept in developing a tissue-engineered hemofilter prototype on a small scale in a rodent model. PMID- 17413559 TI - Removal of bile acids by two different extracorporeal liver support systems in acute-on-chronic liver failure. AB - Acute-on-chronic liver failure (ACLF) is accompanied by marked intrahepatic cholestasis leading to accumulation of cytotoxic bile acids. Extracorporeal liver support systems efficiently remove bile acids, but their effect on bile acid composition in ACLF is unknown. The aim of the present study was to compare elimination of individual plasma bile acids by albumin dialysis (Molecular Adsorbents Recirculating System, MARS) and fractionated plasma separation (Prometheus). Eight consecutive patients with ACLF underwent alternating 6-hour sessions with MARS or Prometheus in a randomized, cross-over design. Serum samples were obtained before, during, and after each treatment, and individual bile acids including cholic acid and chenodeoxycholic acid (CDCA) were measured by gas chromatography. MARS and Prometheus removed total bile acids to a similar extent (reduction ratio, 45% and 46%, respectively). Both devices cleared cholic acid more efficiently than did CDCA. The molar fraction of CDCA (fCDCA) was elevated at baseline and correlated with the degree of liver dysfunction. Prometheus but not MARS treatments further increased fCDCA. Although both devices eliminate total bile acids to a similar extent, clearance of individual bile acids is different, leading to a slight change of the bile acid profile toward hydrophobic bile acids during Prometheus treatments. PMID- 17413560 TI - Comparison of progressive conductivity reduction with diacontrol and standard dialysis. AB - We examined whether progressive reduction of dialysate sodium with Diacontrol (DC, plasma conductivity targeted feedback system) confers any clinical benefit over a similar strategy using dialysis with fixed dialysate conductivity (HD). Ten stable patients entered a randomized crossover study conducted over 360 dialysis sessions. Sodium balance, blood pressure (BP), intradialytic hypotension rates (IDH), thirst score, and extracellular water (ECW) were recorded. Interdialytic ambulatory BP was measured at the highest and lowest conductivities. BP, interdialytic weight gains and thirst scores were low at the outset and were not altered significantly by conductivity reduction. The lowest fixed dialysate setting of 13.2 mS/cm resulted in greater sodium depuration than the lowest conductivity setting allowable with DC, as reflected by lower post dialysis plasma conductivity (13.4 +/- 0.14 mS/cm versus 13.5 +/- 0.04 mS/cm, p < 0.001). Predialysis ECW fell from 0.22 +/- 0.04 l/kg to 0.21 +/- 0.09 l/kg as conductivity reduced with HD (p < 0.05), but did not change significantly with DC. When HD and DC were matched for end-dialysis plasma conductivity, there were no differences in BP, IDH frequency, or dialysis tolerability even at the lowest conductivity settings. In a setting of dialysate sodium reduction, DC did not appear to have any short-term clinical advantage over standard dialysis, and its range is limited at the lower conductivity settings. PMID- 17413561 TI - New ligand coupling procedure for formation of an immunoadsorption wall. AB - To prevent the occurrence of dialysis-related amyloidosis, an immunoadsorption wall based on polyacrylamide has been manufactured by a recently developed, partially incomplete, two-stage polymerization method. During the preparation process, efficient utilization of coupling antibodies is the key to large-scale production of such a toxin removal modality. In this study, we attempted to carry out the ligand coupling procedure after formation of a cyanogen bromide (CNBr) activated stationary phase, using anti-beta2-microglobulin (beta-2M) antibodies. In vitro immunoadsorption tests show that the levels of beta-2M decrease rapidly within the first 2 hours for all the immunoadsorption tests. After that, nearly blank values were reached for tests of initial levels of c. 30 microg/mL and c. 82 microg/mL, whereas a relatively constant level of c. 10 microg/mL was maintained for the test of initial levels of c. 185 microg/mL. The maximum surface binding capacity of the prepared immunoadsorption walls is estimated by fitting experimental data, using a mathematical model of saturation kinetics. The present comparative investigation also suggests the manufacturing process for an immunoadsorption wall could be improved and facilitated by this new ligand coupling procedure without compromising the resulting binding capacity. Furthermore, the experimental protocols as well as the present methodology could be helpful for development of a clinically applicable immunoadsorption wall. PMID- 17413562 TI - Low-volume continuous hemodiafiltration with nafamostat mesilate increases trypsin clearance without decreasing plasma trypsin concentration in severe acute pancreatitis. AB - Continuous hemodiafiltration (CHDF) has recently been used for treatment of severe acute pancreatitis. CHDF is capable of eliminating small molecules from blood, but whether trypsin can be eliminated by CHDF is not clear. In this study, elimination of trypsin-like enzyme activity (TLE) and cationic trypsin-like immunoreactivity (TLI) using low-volume CHDF was examined at the first CHDF session in eight patients with severe acute pancreatitis. CHDF was performed with a polysulfone hemofilter (membrane area, 0.7 m2) and nafamostat mesilate, a protease inhibitor and anticoagulant, at a blood flow rate of 100 ml/min and a filtration and dialysis flow rate of 10 ml/min each. Before beginning CHDF, plasma TLE was 3.41 +/- 2.86 nmol/(ml.min), and TLI was 5,900 +/- 9,008 ng/ml. The average plasma clearances of TLE and TLI achieved by the circuit during the 12-hour therapy were 56.7 +/- 4.9 ml/min and 8.0 +/- 7.2 ml/min, respectively. The average plasma clearance of TLI into the waste fluid was 2.4 +/- 1.6 ml/min whereas TLE was below the measurable sensitivity. The plasma concentration of TLE and TLI remained unchanged. These results indicate that low-volume CHDF using nafamostat mesilate as an anticoagulant can increase trypsin plasma clearance. However, low-volume CHDF is not effective to eliminate the plasma trypsin concentration. PMID- 17413563 TI - Clinical comparison between HeartMate VE auto-mode and HeartMate XVE auto-mode with Opti-Fill and the effect of stroke volume on blood chamber and inflow valve peak pressures. AB - We determined the difference between HeartMate (HM) VE auto mode, average filling 76 mL, and HM XVE Opti-Fill, average filling 79 mL, regarding blood chamber and inflow valve peak pressure pulses (BCPP and IVPP). The relation between stroke volume (SV) and peak pressures was investigated by using a circulatory mock loop. At high SVs, 79 to 83 mL, BCPP and IVPP never exceeded 400 mm Hg. For lower SVs, down to 50 mL, the peak pressures increased to 788 mm Hg for BCPP and 416 mm Hg for IVPP. Distribution of SV was measured in 2 VE and 6 XVE patients during rest and activities of daily living (ADL). For clinical comparison, percentages of SV >78 mL were determined. At rest, 2190 (VE) and 5772 (XVE) pump beats were registered and 4511 (VE) and 8713 (XVE) during ADL. Percentages of "SV >78 mL" at rest, respectively, were 42.5 +/- 3.5 and 78.2 +/- 4.7 (p < 0.01) and during ADL, respectively, 48.7 +/- 7.4 and 73.5 +/- 5.3 (p < 0.01). The Opti-Fill software shows a significant increase in percentage SV >78 mL and makes an important contribution to reducing the incidence of high peak pressures in the clinical setting. PMID- 17413564 TI - Cytoskeletal changes of mesenchymal stem cells during differentiation. AB - Mesenchymal stem cells (MSCs) are progenitors for tissues such as bone and cartilage. In this report, the actin cytoskeleton and nanomechanobiology of human mesenchymal stem cells (hMSCs) were studied using fluorescence microscopy and atomic force microscopy (AFM). Human MSCs were differentiated into chondrocytes and osteoblasts as per previous approaches. Cytochalasin D (CytD) was used to temporarily disrupt cytoskeleton in hMSCs, hMSC-chondrocytes (hMSC-Cys) and hMSC osteoblasts (hMSC-Obs). Fluorescence microscopy revealed a dose-dependent response to CytD. Removal of CytD from the media of cytoskeleton-disrupted cells led to the recovery of the cytoskeletal structures, as confirmed by both fluorescence microscopy and AFM. Force-volume imaging by AFM evaluated the nanomechanics of all three cell types before, during, and after CytD treatment. Cytochalasin D disruption of cytoskeleton had marked effects on hMSCs and hMSC Cys, in comparison with limited cytoskeleton disruption in hMSC-Obs, as confirmed qualitatively by fluorescence microscopy and quantitatively by AFM. Treatment with CytD resulted in morphology changes of all cell types, with significant decreases in the observed Young's Moduli of hMSCs and hMSC-Cys. These data suggest human mesenchymal stem cells alter their cytoskeletal components during differentiation. Additional studies will address the mechanisms of cytoskeletal changes using biochemical and biophysical methods. PMID- 17413565 TI - Anticoagulation of juvenile sheep and goats with heparin, warfarin, and clopidogrel. AB - Little data exist on anticoagulation of young sheep and goats. We tested the effect of heparin, warfarin, and clopidogrel in two sheep and two goats weighing 17-35 kg. Each animal received heparin boluses of 80, 100, and 200 units/kg; goats also received 300, 350, and 400 units/kg. All animals received continuous heparin 40, 60, and 80 units/kg/hour; oral warfarin 0.3, 0.6, and 0.9 mg/kg/day; and oral clopidogrel 75 and 150 mg/day (2.8-3.4 and 5.6-6.9 mg/kg/day). Results were in the form of complete blood counts, activated clotting times (ACT), partial thromboplastin times, prothrombin times, thromboelastograms, and whole blood lumiaggregometry. After heparin boluses of 200 units/kg, sheep and goats reached mean peak ACTs over 400 seconds. After continuous infusions of 40, 60 and 80 units/kg/hour, sheep and goats exceeded our therapeutic range for ACTs (195 215 seconds for sheep, 155-175 seconds for goats). For warfarin therapy, both sheep and goats required treatment with >0.6 mg/kg/day to achieve INRs over 2.5. Clopidogrel treatment, after 14-17 days of 75-150 mg/day, inhibited sheep platelets by 25-36% and goat platelets by 35-46%. We conclude that young sheep and goats can be safely and effectively anticoagulated with heparin and warfarin, and can also show a modest antiplatelet response to clopidogrel. Doses for each drug were generally higher than those used for humans, and warfarin therapy in sheep may be unpredictable. These results should be useful for developing anticoagulation protocols to test pediatric mechanical circulatory support devices. PMID- 17413566 TI - Anoxic ventilation improves systemic perfusion during extracorporeal circulation with uncontrolled systemic-to-pulmonary shunt. AB - Uncontrolled systemic-to-pulmonary shunt results in decreased systemic flow during extracorporeal life support (ECLS). Ligation of systemic-to-pulmonary shunts during ECLS is associated with poor outcome and is not always readily achieved. In ex vivo preparations, alveolar hypoxia results in pulmonary vasoconstriction despite normoxic pulmonary perfusate. We hypothesized that anoxic ventilation would result in reduced pulmonary shunting and increased systemic flow during ECLS in piglets with systemic-to-pulmonary shunt. Four piglets were placed on ECLS with right and left atrial drainage. A shunt was created between the bicarotid trunk and pulmonary artery, using 5-mm ePTFE tubing. Inspired oxygen was reduced to <1% for 10 minutes, then returned to room air; pH, hematocrit, temperature, ventilatory pressures, and total pump flow were maintained constant. Systemic arterial pressure and right atrial return volume and hemoglobin saturation were measured: All decreased significantly upon shunt unclamping. Anoxic ventilation caused increased systemic pressure (34 vs. 28 mm Hg, p < 0.05), flow (335 vs. 278 mL/min, p < 0.05), and systemic venous saturation (53% vs. 48%, p = 0.13) compared with room air ventilation. In conclusion, anoxic ventilation during normoxic ECLS in subjects with systemic-to pulmonary shunts results in a significant and potentially clinically useful reduction in pulmonary shunting. PMID- 17413567 TI - Outcome for pediatric cardiac transplantation with and without bridge methods. AB - Heart transplantation is indicated for children with end-stage heart failure or complex inoperable congenital defects. Due to the shortage of pediatric donor hearts, various bridge techniques have been used for pediatric recipients to prolong patient survival until a heart is available. This study evaluates long term outcome of bridge and nonbridge support for pediatric heart transplantation. Between March 1995 and June 2004, 18 pediatric patients underwent heart transplantation. Six patients (33.3%) underwent biological or mechanical bridge techniques before transplantation. Eight patients (44.4%) required perioperatively extracorporeal membrane oxygenation (ECMO) support. Patient data and records were retrospectively reviewed. Causes of death and long-term outcome were analyzed. Five of eight patients in the ECMO group (62.5%) were successfully decannulated and discharged home with excellent functional classes. No differences in rejection rate, survival rate, and functional class existed between the bridged and nonbridged groups. Overall 1-year and 5-year survival rates were both 83.3% and all have a good functional class. Pediatric heart transplantation can be accomplished with excellent early survival despite multiple prior cardiac operations and relatively severe illness. For the variety in small, low-body-weight pediatric patients, mechanical circulatory support using ECMO is suitable for managing sudden collapse while waiting for heart transplantation, and graft dysfunction after cardiac transplantation. The mortality rate is acceptable in this very high-risk group of patients and long term outcome is good. PMID- 17413568 TI - Extracorporeal membrane oxygenation for perioperative cardiac support in children I: experience at the Deutsches Herzzentrum Berlin (1987-2005). AB - We report our experience in pediatric patients supported by extracorporeal membrane oxygenation (ECMO) for perioperative circulatory failure from January 1987 to June 2005. Pediatric patients (n = 110) who had ECMO support for congenital heart defects, myocarditis, and cardiomyopathy (age range, newborn to 18 years; weight range, 2.3-69 kg) were included and divided into three groups based on timing of ECMO support. EMCO support was used preoperatively in 21 patients (19.1%) (mean age, 4 years +/- 8 months; mean weight, 23.7 +/- 8.9 kg). Duration of ECMO support was 8.3 +/- 7 days. Fifty-six patients (56.56%) (mean age, 5.11 +/- 5 years; mean weight, 15.7 +/- 6.9 kg) had intraoperative ECMO support for myocardial insufficiency, low output syndrome, right ventricular failure, left ventricular failure, malignant arrhythmia, pulmonary hypertension, and repeated resuscitation. Mean duration of ECMO support was 4.98 +/- 1 days. Postoperative ECMO support was used in 29 patients (mean age, 7.5 +/- 1 years; mean weight, 23.4 +/- 6.4 kg). Mean duration of ECMO was 4.6 +/- 1 days. Mean postoperative day of ECMO institution was 40.4 +/- 2 days. Our experience shows that ECMO support can be offered perioperatively to any patient with potentially reversible pulmonary, cardiac, or cardiopulmonary failure, excluding those whose outcome is inevitable. PMID- 17413570 TI - Letter to the editor: A possible major mistake in the paper entitled "collected nondimensional performance of rotary dynamic blood pump": Smith WA, Allaire P, Antaki J, Butler KC, Kerkhoffs W, Kink T, Loree H, Reul H. ASAIO Journal 50: 25 32, 2004. PMID- 17413571 TI - Contribution of F-18 FDG PET-CT in the detection of systemic spread of primary central nervous system lymphoma. AB - PURPOSE: Primary central nervous system lymphoma (PCNSL) accounts for approximately 3% of all primary brain tumors and 1% of all non-Hodgkin lymphomas. Detection of systemic spread of PCNSL, although rare (4%), is very important since therapy is usually modified. Contrast-enhanced computed tomography (CT) is commonly used for systemic staging of PCNSL. No previous case report is available in the published literature elaborating the potential contribution of F-18 FDG PET in systemic staging of PCNSL. The purpose of this case report was to document the potential usefulness of F-18 FDG-PET in the detection of occult systemic involvement in PCNSL. MATERIALS AND METHODS: A 50-year-old, immunocompetent, male patient completed successful treatment of PCNSL. As part of a routine pretransplant evaluation he had an F-18 FDG PET coregistered with CT (PET-CT). The PET-CT results were then compared with those of contrast-enhanced CT of the chest, abdomen, and pelvis. RESULTS: The PET-CT examination detected multiple sites of extranodal systemic disease that were not seen in the contrast-enhanced CT of the chest, abdomen, and pelvis (both studies were performed within 24 hours of each other). Percutaneous ultrasound guided biopsy confirmed the presence of systemic spread of PCNSL. The patient's subsequent therapy was modified to include rituximab with cyclophosphamide, doxorubicin, vincristine, prednisone (R CHOP). A follow up PET-CT confirmed resolution of systemic spread. CONCLUSION: F 18 FDG PET coregistered to CT may be a useful examination in the detection and monitoring for systemic spread of the disease in PCNSL patients. PMID- 17413572 TI - Positron emission tomography for predicting malignancy of sinonasal inverted papilloma. AB - BACKGROUND: Sinonasal inverted papilloma (IP) is an uncommon nasal tumor. It is known for being locally invasive and recurrent, with a high probability of malignant transformation. Accurate initial staging and close surveillance are critical in the management of the disease. METHODS AND RESULTS: Fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) was performed in 5 patients with IP. High FDG uptake (maximum standardized uptake (SUVmax) value >3.0) was observed in all patients, and 2 patients with IP associated with squamous cell carcinoma (SCC) exhibited higher SUVmax values. CONCLUSION: The SUVmax value of a sinonasal tumor can warn the surgeon of the probability of an associated malignancy, even when preoperative biopsy demonstrates a purely benign papilloma. PMID- 17413573 TI - Treatment for microcarcinoma of the thyroid--clinical experience. AB - AIMS AND BACKGROUND: Papillary microcarcinoma of the thyroid is a specific subgroup of papillary thyroid carcinoma (PTC) and account for up to 30% of all PTC. According to the World Health Organization, papillary microcarcinoma of the thyroid (PMC) is defined as a papillary thyroid carcinoma measuring /=25 predicted 30-day mortality with a sensitivity of 74.19%, a specificity of 90.91%, and an accuracy of 78.58%. The positive predictive value was 95.83% and the negative predictive value 55.56%. CONCLUSIONS: Patients with cirrhosis who are not candidates for liver transplantation are referred to hospice care at a late stage with an average survival of 1 month. The MELD score correlates with survival of cirrhosis patients enrolled in hospice and can be used to estimate 30-day mortality. Further, research is needed to determine a MELD score that predicts a survival of 6 months or less, an important determinant of appropriate hospice referrals. PMID- 17413613 TI - A multivariable model of clinical variables predicts advanced fibrosis in chronic hepatitis C. AB - BACKGROUND: A noninvasive method to identify advanced hepatic fibrosis (AHF) in chronic hepatitis C (CHC) could preclude the need for routine liver biopsy. Recent evidence suggests that obesity may contribute to hepatic fibrosis in hepatitis C virus infection. GOALS: To determine whether clinical variables, including body mass index (BMI), can predict risk of AHF. STUDY: Retrospective review of untreated CHC patients evaluated between 1993 and 2002 without clinical or physical evidence of end-stage liver disease. Liver biopsies were scored for fibrosis, steatosis, and inflammation. Multivariable analysis was used to derive and internally validate a prediction equation. A clinical index was created from the equation by assigning points for each variable. The risk of AHF was measured for each risk category. RESULTS: Two hundred eighty-six satisfied inclusion criteria, of which 86 (30%) had AHF. In the derivation subgroup (N=190), 5 factors were independently associated with AHF: diabetes mellitus, platelets count <150,000, aspartate aminotransferase > or =65 IU/mL, international normalized ratio > or =1.1, and bilirubin > or =0.85 mg/dL. The corresponding risk index contained 3 categories: low-risk (score of 0), intermediate risk (scores of 1 to 3), and high risk (scores of > or =4), in which the respective risks of AHF were 9%, 34%, and 92%. Inclusion of BMI did not improve model performance. CONCLUSIONS: A model for estimating AHF risk in CHC performed well in this population. BMI had no effect on the risk of AHF. If this model can be validated in other patient cohorts, it could preclude the need for liver biopsy in patients with scores of 0 or > or =4. PMID- 17413614 TI - Predicting pancreas divisum by inspection of the minor papilla: a prospective study. AB - BACKGROUND: Minor papilla (MP) cannulation remains difficult in some patients despite improved cannulation devices and techniques. Therefore, noninvasive methods to predict presence of pancreas divisum (Pdiv) are desired. If Pdiv could be recognized before any cannulation is attempted, appropriate attention could be focused on MP cannulation and prolonged major papilla attempt at pancreatogram can be avoided. AIM: To evaluate whether simple inspection of the MP during initial duodenal entry during endoscopic retrograde cholangiopancreatography (ERCP) can predict the presence of Pdiv. METHODS: Patients undergoing ERCP with intended pancreatic duct cannulation were included in the study. Assessment of the MP was made during a 30 to 60-second period of inspection including aspiration of duodenal luminal air with the endoscope. The probability of Pdiv based on the appearance of the MP was then estimated by assessing the size of the MP, the diameter of MP orifice, and whether juice was seen draining through the MP orifice. RESULTS: Two hundred and twelve patients were prospectively analyzed. The most common indications for ERCP were suspected sphincter of Oddi dysfunction (44.3%) and idiopathic pancreatitis (34.4%). Pancreatograms revealed Pdiv in 42 (19.8%). The MP appearance had 54.7% sensitivity and 90% specificity in detecting Pdiv (positive predictive value 57.5%, negative predicting value 88.9%). Seventy percent of the false positive patients had an obstructing pathology at the ventral duct that would explain the MP appearance. CONCLUSIONS: Certain features of the MP (enlarged papilla, open orifice) had a moderate predictive value for the presence of Pdiv or an obstruction at the major papilla. However, a significant number of patients with Pdiv did not have these features. PMID- 17413615 TI - Risk for post-ERCP pancreatitis after needle knife precut sphincterotomy following repeated cannulation attempts. AB - GOALS: The aim of this study was to determine the risk and identify the factors associated with post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis in patients who undergo needle knife precut sphincterotomy (NKS). In addition, we evaluated the effect of gabexate for the prevention of post-ERCP pancreatitis. BACKGROUND: NKS, after repeated cannulation attempts during ERCP, is known to increase the risk of post-ERCP pancreatitis. However, the specific risk factors for post-ERCP pancreatitis have not been identified, and the preventive role of protease inhibitors, such as gabexate, has not yet been established. STUDY: The medical records of 200 patients who underwent NKS, after repeated cannulation attempts during ERCP, were reviewed retrospectively. The potential risk factors for post-ERCP pancreatitis were investigated. The effect of gabexate infused after the ERCP procedure was also evaluated. RESULTS: Thirteen (6.5%) patients out of 200 developed post-ERCP pancreatitis. Sex, age, the presence of pancreatitis at procedure, underlying disease, direction of the sphincterotomy, success or failure of cannulation after NKS, diameter of common bile duct, pancreatic duct status, and the presence of acinar filling were not associated with the risk of pancreatitis. Gabexate infusion after ERCP increased the incidence of ERCP-associated pancreatitis. CONCLUSIONS: We could not identify any risk factors associated with the development of post-ERCP pancreatitis in patients who underwent NKS after repeated cannulation attempts during ERCP. However, gabexate administered after the ERCP procedure was found to increase the incidence of pancreatitis. PMID- 17413616 TI - Severe allergic eczema due to pegylated alpha-interferon may abate after switching to daily conventional alpha-interferon. PMID- 17413617 TI - Urgent awareness needed to combat Asia's hidden epidemic Hepatitis C. PMID- 17413619 TI - Clinical and imaging characteristics of orbital leukemic tumors. AB - PURPOSE: To characterize the clinical and imaging features of orbital leukemic tumors in 27 patients seen and treated at St. Jude Children's Research Hospital. METHODS: A retrospective review was performed on the clinical and imaging records of 27 consecutive patients with a diagnosis of orbital leukemic tumors. The following data were extracted from the patients' records: age at diagnosis of orbital leukemic tumors, sex, race, national origin, type of leukemia, temporal relationship of orbital disease to diagnosis of leukemia, survival from diagnosis of orbital leukemic tumor, laterality of the orbital disease, location of the mass within the orbit, imaging features of the mass, chemotherapeutic protocol, treatment with bone marrow transplant, and orbital radiation. RESULTS: The median age at diagnosis of orbital leukemic tumor was 8 years (range, 1-18 years). Nineteen of the 27 patients were male, and 21 patients were born and lived in the United States. Twenty-one patients had acute myeloid leukemia, five had acute lymphoblastic leukemia, and one had chronic myelogenous leukemia. In 85% of patients (n = 23), the diagnosis of leukemia was based on the bone marrow examination findings. Orbital imaging revealed homogenous masses that molded to one or more orbital walls without bony destruction. Nine patients had bilateral orbital involvement. All patients received multiagent systemic chemotherapy, and 14 underwent bone marrow transplantation. Five patients received external beam radiation for the treatment of orbital disease. Fifteen (55.6%) of the 27 patients were alive at the time of the study. The median survival for all patients was 4.75 years (range, 0.1-24 years) after the diagnosis of orbital disease. CONCLUSIONS: Orbital leukemic tumors occur most commonly in the first decade of life, in association with acute myeloid leukemia. They appear as homogenous masses along the orbital walls. Although the overall survival rate for patients with leukemia has improved over the past 3 decades, the mortality of patients who develop orbital leukemic tumors remains high. PMID- 17413620 TI - Prognosis for patients in a Korean population with ocular adnexal lymphoproliferative lesions. AB - PURPOSE: To analyze the clinical features, treatment outcomes, and prognostic factors associated with lymphoproliferative lesions of the ocular adnexa in a Korean population. METHODS: Data from 69 patients treated for adnexal lymphoproliferative disease in the 12-year period from 1991 to 2002 were retrospectively evaluated. RESULTS: The 69 patients had a median age of 46 years (range, 15-73 years); 60 of these patients were diagnosed with extranodal marginal zone B cell lymphomas (MALT lymphomas) and had 10 year cause specific survival and relapse free survival rates of 95.6% and 82.6%, respectively. Of 6 patients (8.7%) with concurrent systemic lymphoma, including 4 diagnosed with stage IV disease, 3 died from lymphoma. Only one patient with a primary ocular adnexal MALT lymphoma developed systemic lymphoma, which was treated with surgical resection. Local recurrence either at the primary site or in the fellow eye occurred in 11.6% of patients at a median follow-up time of 102 months (range, 79-132 months), and was controlled using repeat irradiation in all cases. Statistical analysis showed the presence of concurrent systemic lymphoma, bilateral disease, and an advanced stage at diagnosis, were linked to lymphoma related death (Log-rank test, p < 0.05) and systemic progression (Fisher's exact test, p < 0.05), and that the tumor location was not a prognostic factor for lymphoma-related death or relapse at any site. CONCLUSIONS: Ocular adnexal lymphoproliferative disease in Koreans occurred at a relatively young age, and was mostly orbitally located and of the MALT subtype, which is highly localized and rarely associated with extraorbital relapse. Primary or secondary status, stage at presentation, and bilaterality were found to be prognostic factors. PMID- 17413621 TI - Late-onset presentation of orbital lymphangioma. AB - PURPOSE: To report a group of patients with biopsy-proven orbital lymphangioma diagnosed late in life. METHODS: The clinical records and histopathologic findings of 3 patients with biopsy-proven orbital lymphangioma were reviewed. RESULTS: Three patients were identified who were diagnosed with orbital lymphangioma at the ages of 70, 80, and 82 years old. Each patient presented with different symptoms and signs including presence of a superficial mass, periorbital pain, and gradual painless proptosis, respectively. In one case, imaging revealed an isolated superficial mass, and in the other 2 cases it demonstrated the presence of a heterogeneous, cystic, retrobulbar mass. Pathologic findings in all cases included endothelium-lined channels with intervening fibrous septae infiltrated with aggregates of lymphocytes. Two of the three patients had complete excisions without recurrences and the third patient was followed after biopsy. CONCLUSIONS: Lymphangioma, one of the most common vascular lesions in childhood, should also be included in the differential diagnosis of orbital masses in adults, including elderly adults. PMID- 17413622 TI - Association between thyroid autoimmune dysfunction and non-thyroid autoimmune diseases. AB - PURPOSE: To assess the prevalence of the association between thyroid autoimmune dysfunction in patients with and without Graves orbitopathy and non-thyroid autoimmune diseases. METHODS: Retrospective review of the medical records of 254 consecutive patients with thyroid autoimmune disease with (n = 150) and without (n = 104) orbitopathy who had been followed at the same institution by ophthalmologists and general clinicians. All medical records contained information on any systemic diseases of the patients and a detailed description of their eye examinations. The mean follow-up period was 5.25 +/- 4.67 years. RESULTS: Non-thyroid autoimmune diseases were detected in 24 (9.4%) patients. Type 1 diabetes was the most prevalent non-thyroid autoimmune disease diagnosed in the patients without orbitopathy (7 patients, 6.7%). For the patients with orbitopathy, vitiligo was the most prevalent condition, affecting 6 patients (4%). Other diseases including systemic sclerosis, systemic lupus erythematosus, myasthenia gravis, Sjogren syndrome, and rheumatoid arthritis were seen in a few patients in both groups. The time intervals between the diagnoses of the orbitopathy and the non-thyroid autoimmune disease were highly variable, ranging from none (concomitance of the 2 conditions) to decades. CONCLUSIONS: The present data show that several non-thyroid autoimmune diseases may be associated with thyroid autoimmune dysfunction. Patients with Graves disease without orbitopathy are likely to develop polyglandular syndrome due to the occurrence of type 1 diabetes. Patients with Graves orbitopathy should be screened for other autoimmune conditions, especially vitiligo. PMID- 17413623 TI - Optic neuropathy associated with botulinum A toxin in thyroid-related orbitopathy. AB - PURPOSE: To report the development of optic neuropathy after botulinum A toxin injection for restrictive myopathy from thyroid-related orbitopathy (TRO). METHODS: We retrospectively reviewed the records of three patients with TRO who underwent botulinum A toxin injection for restrictive myopathy and subsequently developed optic neuropathy. Development of optic neuropathy was measured by visual acuity, color vision testing, visual field testing, and relative afferent pupillary testing. RESULTS: At 3 week follow-up after botulinum A toxin injection, three patients were noted to have clinical signs and symptoms of optic neuropathy in the ipsilateral eye following injection of botulinum A toxin for restrictive myopathy. Treatment with oral steroids followed by orbital wall decompression reversed the optic neuropathy. CONCLUSIONS: To our knowledge, this is the first report of optic neuropathy associated with botulinum A toxin injection in TRO. Clinicians should be aware of this potential vision threatening complication. PMID- 17413624 TI - Orbital fractures complicated by late enophthalmos: higher prevalence in patients with multiple trauma. AB - PURPOSE: To present our experience with orbital fracture treatment outcomes in patients with multiple trauma and those suffering localized orbital trauma. METHODS: Retrospective data review of all patients examined for ocular motility problems and/or enophthalmos following orbital trauma in a 4-year period. RESULTS: Forty-three patients were included in the study: 31 (72%) had localized orbital trauma (LOT) and 12 (28%) had concomitant traumatic insults to other organs (MT). More orbital walls were affected in MT patients than in LOT patients, and the incidence of zygomatic fracture was higher in MT patients. Late enopthalmos was much more prevalent in the MT group compared with the LOT group. Differences of outcome of extra ocular motility disturbance between groups in our series did not reach statistical significance. CONCLUSIONS: The findings of more walls affected and higher incidence of zygomatic fractures in MT patients probably represent a stronger impact of the original insult, causing both more damage to other organs and more severe damage to the orbit. The increased rate of late enophthalmos in MT patients may be associated with their primary presentation to the emergency room with potentially life-threatening injuries. Under such circumstances, thorough ophthalmologic examination is nearly impossible, both because the patient cannot cooperate sufficiently and because medical priorities dictate concentration on taking care of the injuries threatening life. The lack of a thorough ophthalmic examination prevents early comprehensive treatment. PMID- 17413625 TI - Assessment of the predictive value of intraoperative eyelid height measurements in sitting and supine positions during blepharoptosis repair. AB - PURPOSE: To examine relationships between intraoperative and postoperative eyelid heights in relation to patient postures in blepharoptosis surgery. METHODS: Thirty eyelids in 15 patients with bilateral aponeurotic ptosis were selected. The margin reflex distance-1 (MRD-1) was measured with the patients in the sitting position preoperatively, intraoperatively, and in the postoperative 6 weeks. MRD-1 also was measured intraoperatively with patients in the supine position. The intraoperative sitting and supine values and the postoperative values were statistically compared using a paired t test, and lateralities in those groups were statistically analyzed using Student's t test. RESULTS: From the intraoperative measurements, mean MRD-1 values in the sitting position (right: 3.80 mm, left: 3.73 mm) were significantly lower than those in the supine position (right: 4.37 mm, p < 0.001; left: 4.33 mm, p < 0.001). In the postoperative 6 weeks, mean MRD-1 values (right: 3.80 mm, left: 3.63 mm) were not significantly different from intraoperative values in the sitting position (right: p = 1.000, left: p = 0.189), but were significantly lower than intraoperative values in the supine position (right: p < 0.001, left: p < 0.001). There was no significant laterality among the three groups (supine intraoperative: p = 0.890, sitting intraoperative: p = 0.785, postoperative: p = 0.546). CONCLUSION: Intraoperative measurements in the sitting position more accurately predicted postoperative eyelid height than did measurements with the patient in a supine position. PMID- 17413626 TI - Ultrasound biomicroscopy of the upper eyelid structures in normal eyelids. AB - PURPOSE: To evaluate the normal upper eyelid structures quantitatively and qualitatively using ultrasound biomicroscopy (UBM). METHODS: Sixteen upper eyelids of 16 healthy subjects with no eyelid problems were evaluated with UBM. The orbicularis oculi muscle, levator aponeurosis, Muller muscle-conjunctival complex, and tarsus were imaged centrally just above the tarsus, and the tarsus was imaged just above the eyelid margin. The thickness of these structures was measured primary gaze. The measurements of levator aponeurosis and Muller muscle conjunctival complex were repeated in upgaze. RESULTS: Of 16 patients, 8 were men and 8 were women. Eight were more than 40 years old and eight were younger. Orbicularis oculi muscle and Muller muscle-conjunctival complex were echo-dense, while levator aponeurosis and tarsus were echo-lucent. The mean thickness was 0.74 +/- 0.11 mm for orbicularis oculi muscle, 0.44 +/- 0.67 mm for levator aponeurosis, 0.91 +/- 0.15 mm for tarsus, and 0.38 +/- 0.64 mm for Muller muscle conjunctival complex. There was no significant difference in the thickness of these structures between the patients younger than 40 years old and older patients, or between male and female patients (p > 0.05). In upgaze, the increases in thickness of levator aponeurosis and Muller muscle-conjunctival complex were a mean of 53% and 32%, respectively. CONCLUSIONS: UBM is an easy, non-invasive test used to visualize normal eyelid structures. It can be used in the diagnosis and follow-up of eyelid problems. The normal upper eyelid structures showed no significant differences based on age or gender. PMID- 17413627 TI - Facial nerve paralysis: an unrecognized cause of lower eyelid entropion in the pediatric population. AB - PURPOSE: To describe the association between entropion and pediatric facial nerve paralysis in the pediatric population. METHODS: A retrospective case series was collected from 5 pediatric patients who required surgical correction for symptomatic entropion with a history of facial nerve palsy. RESULTS: All 5 patients presented with epiphora and punctate keratopathy. Following surgical correction of the entropion, all ocular symptoms resolved. CONCLUSIONS: To the best of our knowledge, this is the first report linking facial nerve paralysis with entropion in the pediatric population. Ophthalmologists and pediatricians should consider this in the evaluation of children with craniofacial abnormalities and facial nerve paralysis. PMID- 17413628 TI - Efficacy of skin cooling and EMLA cream application for pain relief of periocular botulinum toxin injection. AB - PURPOSE: To investigate the efficacy of EMLA cream and local dry cold application for pain relief before periocular botulinum toxin injection, and to compare these two methods. METHODS: In this prospective study, 40 patients underwent bilateral periocular botulinum injections for blepharospasm treatment or wrinkle reduction. Patients were divided into three treatment groups. In the first group (n = 12), cold was applied to the periocular area on one side before injection; the other side served as the control. In the second group (n = 12), topical anesthetic cream (EMLA) was applied to one side and the other side served as the control. In the third group (n = 16), cold was applied to one side and EMLA was applied to the other side. A visual analog scale was used for pain intensity, and patients in the third group were asked which treatment they preferred. RESULTS: In the first group, the average pain score on the side where cold was applied was 3.00 +/- 1.70, whereas it was 5.83 +/- 1.40 on the control side (p < 0.001). In the second group, the average pain score on the side receiving EMLA was 3.25 +/- 1.86, and on the control side was 5.83 +/- 1.89 (p < 0.001). In the third group, the average pain score was 3.18 +/- 1.68 for the EMLA side and 3.12 +/- 1.31 for the cooled side (p > 0.05); nine of the 16 patients (56.2%) preferred EMLA. CONCLUSIONS: Skin cooling and EMLA applications significantly decrease the pain associated with periocular botulinum toxin injections. Clinically or statistically significant difference in pain scores between the two methods was not noted. Patients had a slight preference for EMLA cream over skin cooling. PMID- 17413629 TI - Use of the diode laser with intraoperative mitomycin C in endocanalicular laser dacryocystorhinostomy. AB - PURPOSE: To determine the safety and efficacy of the diode laser with intraoperative mitomycin C in endocanalicular laser dacryocystorhinostomy (ECL DCR). METHODS: In a prospective case series of 40 ECL-DCRs using the diode laser, mitomycin C was placed intraoperatively in all cases. The main outcome measure was resolution or improvement of epiphora and no major laser damage intranasally. Patients were followed for at least 18 months. RESULTS: Forty consecutive ECL DCRs on 30 patients (23 females, 7 males, mean age 62 years) were performed from April 2000 to December 2001. The success rate at 12 months postoperatively was 87.5%. All failures were due to a constricted nasal osteotomy. No significant intraoperative or postoperative complications were recorded. CONCLUSIONS: Diode laser ECL-DCR with mitomycin C appears to be a safe and effective treatment modality for primary acquired nasolacrimal duct obstruction. PMID- 17413630 TI - Lodged intracanalicular plugs as a cause of lacrimal obstruction. AB - PURPOSE: To evaluate the complications and success of treatment of migrated or lodged intracanalicular and punctal plugs. METHODS: A retrospective chart review of all cases having either a dacryocystorhinostomy or surgical removal of an intracanalicular or punctal plug from 1992 to 2006, in a single physician referral oculoplastics practice, was performed to identify cases in which a retained lacrimal plug required surgical intervention. Patients presented with symptoms of tearing, infection, or granuloma formation. The charts of 998 surgical cases were reviewed, from which 66 eyes (6.6%) were determined to have had lodged lacrimal plugs that required surgical removal, thus qualifying them for inclusion in this study. Patients were followed after surgery until reconstructive silicone tubing was removed (range, 6 weeks to 6 months), and each patient was questioned regarding symptoms. A comparison group of 336 eyes that had collared punctal plugs placed served as the control group. RESULTS: All cases were noted to have complications from intracanalicular plugs. No complications were noted from other forms of lacrimal plugs. All eyes in this series required a canaliculotomy or a dacryocystorhinostomy after office irrigation failed to dislodge the plug. Five eyes presented with canaliculitis, 28 eyes presented with epiphora, and 29 eyes presented with dacryocystitis. Four of 66 eyes (6%) in this cohort presented with a pyogenic granuloma. Five eyes (8%) presented with canaliculitis. Forty-nine of 66 eyes (74%) were asymptomatic following treatment, with no observable infection or epiphora. Seven of 66 eyes (11%) had some improvement in symptoms and another 10 of 66 eyes (15%) had no change in symptoms after treatment. No complications requiring surgical intervention were encountered in the control group of collared punctal plugs. CONCLUSIONS: Intracanalicular-type plugs may lodge in the lacrimal outflow system. This may result in epiphora, canaliculitis, or dacryocystitis that may require major reconstructive surgery. Despite surgical intervention, these symptoms do not always resolve (26% of eyes in this study had persistent epiphora). Intracanalicular plugs were observed to be associated with a higher rate of granulation tissue formation in the lacrimal outflow tract when compared with other forms of punctal plugs. As a result of the increased number of complications seen with intracanalicular plugs, caution is advised with respect to use of these devices. The relative infrequency of complications seen with collared punctal plugs suggests a safer alternative. Data from this study lead the authors to advocate the consideration of other forms of lacrimal occlusion due to the high number of complications noted with intracanalicular plugs, and the availability of other reversible forms of punctal occlusion. PMID- 17413631 TI - Composite lymphoma of the orbit treated with rituximab. AB - We describe an unusual case of composite lymphoma (CL) in the orbit. The clinical history and biopsy specimen of an 82-year-old woman with a right orbital mass were evaluated. The orbital biopsy contained a dense lymphocytic infiltration and nodules of large lymphocyte that immunostained positive for CD20. Flow cytometric immunophenotyping showed two distinct populations of cells, confirming the diagnosis of a simultaneous follicular lymphoma (FL) and chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). The patient was treated with CD20 antibody (rituximab) and is in remission at 3 months follow-up. PMID- 17413632 TI - Fibrous histiocytoma of the lacrimal gland. AB - Fibrous histiocytoma, a primary mesenchymal tumor of the orbit, is known to arise from various ocular and adnexal tissues. We are unable to find a published report of this tumor originating from the lacrimal gland. We report a case of a benign fibrous histiocytoma of the lacrimal gland in an 11 year old girl who presented with painless, progressive eyelid swelling and mild proptosis. Imaging studies revealed a discrete mass in the lacrimal gland region. The tumor was completely excised by anterolateral orbitotomy. Light microscopy showed a spindle cell tumor arising from the lacrimal gland. The tumor cells were arranged in a characteristic storiform (cartwheel) pattern with no pleomorphism or mitotic figures. Immunohistochemically, the tumor cells were focally positive for CD-68 and negative for S-100, smooth muscle actin, vimentin, and CD-34, which ruled out neurofibroma, leiomyoma, solitary fibrous tumor, and hemangiopericytoma. Based on these features, a diagnosis of benign fibrous histiocytoma was made. PMID- 17413633 TI - Orbital embryonal rhabdomyosarcoma in association with neurofibromatosis type 1. AB - A 20-month-old child with systemic features of neurofibromatosis type 1 presented with sudden-onset proptosis. Biopsy and histopathology confirmed the diagnosis of an embryonal rhabdomyosarcoma. The tumor regressed completely with chemotherapy and external beam radiotherapy. This case highlights the association of rhabdomyosarcoma with neurofibromatosis type 1. PMID- 17413634 TI - Alveolar rhabdomyosarcoma presenting as an acute orbital mass in the medial rectus muscle. AB - Rhabdomyosarcoma is the most common pediatric primary neoplasm in the orbit, often presenting with rapid proptosis and orbital symptoms. We describe a 15-year old girl who presented with an acute mass in her medial rectus muscle that was subsequently diagnosed as widely disseminated alveolar rhabdomyosarcoma. To our knowledge, this represents the first reported case in which an enlarged extraocular muscle was the initial manifestation of disseminated alveolar rhabdomyosarcoma. PMID- 17413635 TI - Inverted papilloma of the nasolacrimal system invading the orbit. AB - A 73-year-old man with a history of epiphora, discharge from the left eye, and left-sided nasal congestion underwent external dacryocystorhinostomy for nasolacrimal duct obstruction. The procedure revealed bony erosion of the majority of the lacrimal sac fossa and a large papillomatous mass filling the lacrimal sac and nasolacrimal duct. Inverted papilloma was diagnosed via biopsy. A subsequent orbitotomy with combined endoscopic medial maxillectomy was performed to remove the mass. This case illustrates the importance of including inverted papilloma, a benign but invasive neoplasm, in the differential diagnosis of nasolacrimal duct obstruction. PMID- 17413636 TI - Primary Ewing sarcoma of the orbit in an adult. AB - A 42-year-old man presented with visual field loss in the right eye and unintentional weight loss associated with a right orbital mass. Surgical resection and biopsy revealed a diagnosis of Ewing sarcoma, and systemic evaluation showed no metastasis. Subsequent chemotherapy and radiation resulted in tumor resolution without recurrence for 3.5 years. Although systemic symptoms are associated with a poorer prognosis, longer survival may be possible in the absence of metastasis. PMID- 17413637 TI - Orbital metastases from male breast cancer in two cases. AB - We present 2 cases of orbital metastases of male breast cancer, a very rare association in the context of a rare disease. We also provide a review of the literature on this topic including the 4 previously reported cases. Our cases presented with diplopia and proptosis. Orbital biopsies were performed that revealed metastases of infiltrative ductal carcinoma (ICD). PMID- 17413638 TI - Traumatic subluxation of the globe into the maxillary sinus. AB - Fractures of the orbit most commonly involve the orbital floor, and enophthalmos is a common sign of orbital fractures. Complete globe dislocation in the maxillary sinus, however, has rarely been reported. Herein we report on an 18 year-old man referred to us 1 week following a car accident. At presentation, the globe was not seen within the right orbit. CT revealed a large orbital floor fracture with globe dislocation in the right maxillary sinus. The patient underwent repair of the fracture using a Medpor implant together with repositioning of the globe. Preoperative vision was no light perception, which remained unchanged during follow-up. PMID- 17413639 TI - Avulsion of the optic nerve with luxation of the eye after motor vehicle accident. AB - A 27-year-old man suffered simultaneous avulsion of the optic nerve and luxation of an intact globe in a motor vehicle accident. CT was not diagnostic of the nerve avulsion. The injury was addressed by emergent replacement of the globe within the orbit. The patient obtained psychological benefit by retaining the eye initially, but required subsequent enucleation for prolonged ocular inflammation. Initial repositioning of intact, luxated globes offers potential cosmetic and psychological benefits. If necessary, enucleation can be performed later. PMID- 17413640 TI - Methicillin-resistant Staphylococcus aureus caruncle abscess. AB - A 60-year-old woman under treatment for a left lower eyelid abscess developed a right caruncle abscess. Methicillin-resistant Staphylococcus aureus (MRSA) was identified in cultures performed on both lesions. Treatment consisted of oral ciprofloxacin and moxifloxacin drops. There was resolution of both lesions with incision and drainage, and antibiotic therapy. MRSA is a pathogen that can be readily isolated from the caruncle. The authors are unaware of previous reported cases of a MRSA caruncle abscess. PMID- 17413641 TI - Persistent periorbital and facial lymphedema associated with Group A beta hemolytic streptococcal infection (erysipelas). AB - Chronic lymphedema is both a risk factor for and consequence of erysipelas (cellulitis). We report a case of a 62-year-old woman with rheumatoid arthritis treated with etanercept and prednisone, who developed chronic periorbital lymphedema 2 months after Group A beta-hemolytic streptococcus infection of the face. She had significant ptosis OS and thickened, hyperpigmented periorbital skin. Biopsies were consistent with chronic lymphedema. Of note, on 6 months follow-up, the patient's appearance was improved though she still had residual ptosis. A period of extended observation may be warranted in these cases. PMID- 17413642 TI - Bilateral eyelid edema and orbital cellulitis associated with Pott's puffy tumor. AB - Pott's puffy tumor is rarely associated with orbital cellulitis. We describe a pediatric patient with Pott's puffy tumor and right-sided orbital cellulitis. He underwent urgent surgical drainage of a glabellar subperiosteal abscess and completed a six week course of intravenous antibiotics with complete resolution of infection and no long-term sequelae. Close monitoring with imaging for intracranial spread and multidisciplinary surgical intervention are commonly recommended. PMID- 17413643 TI - Septic pulmonary embolism associated with orbital abscess and sinusitis. AB - A 13-year-old white boy was admitted to an outside institution for antibiotic treatment of right preseptal cellulitis. He subsequently developed a right orbital abscess, which could not be located on surgical exploration. On postoperative day 2, he developed acute chest pain and was transferred to Children's Hospital of Michigan for evaluation and treatment. CT of the thorax demonstrated multiple, bilateral lower lobe lesions consistent with septic pulmonary embolism. The right orbit was re-explored with successful drainage of an intraconal orbital abscess. He was treated with intravenous antibiotics with subsequent improvement of the orbital cellulitis and chest pain. He was discharged home on intravenous antibiotics. At 1 month, follow-up chest CT demonstrated near resolution of the lung lesions. PMID- 17413644 TI - Perilimbal needle manipulation of conjunctival chemosis after cosmetic lower eyelid blepharoplasty. AB - We report an effective treatment procedure for postoperative conjunctival chemosis following cosmetic lower eyelid blepharoplasty. A minimally invasive procedure was used to treat three patients who presented with chronic conjunctival chemosis following lower eyelid blepharoplasty. This procedure utilizes local anesthesia and perilimbal manipulation with a 27-gauge needle. Two months following the procedure, reduction of the chemosis was observed in all three patients. Our cases indicate that perilimbal manipulation with a 27-gauge needle is a viable treatment for this surgical complication. PMID- 17413645 TI - Unilateral eyelid lag and retraction as sequelae of trachoma. AB - A 42-year-old woman had a large right palpebral fissure 13 mm high with 0.5 mm of scleral show both superiorly and inferiorly due to upper and lower eyelid retraction. The upper eyelid showed eyelid lag behind the globe only during the initial ocular movement. Superior tarsal height measured 7.5 mm with palpebral conjunctival cicatrization and Arlt's line. Severe cases of trachoma may lead to contracture of the conjunctiva and deeper tissues including Muller muscle and the tarsal plate, which supports the insertion of the levator aponeurosis. Shortening of the posterior lamella and fornix contracture would explain eyelid retraction and failure of the upper eyelid to smoothly follow the globe on downgaze. PMID- 17413646 TI - Upper and lower system nasolacrimal duct stenosis secondary to paclitaxel. AB - A 66-year-old man underwent chemotherapy and radiation for angiosarcoma of the head and neck. Paclitaxel was used in his chemotherapeutic regimen. Two months post treatment with paclitaxel, he developed epiphora. He was found to have severe punctal and canalicular stenosis at the time of surgery. PMID- 17413647 TI - A novel approach to lacrimal sac dissection in dacryocystorhinostomy. AB - The use of membrane-peeling vitreoretinal forceps to facilitate lacrimal sac dissection in external dacryocystorhinostomy is demonstrated. The closed vitreoretinal forceps are advanced via the upper punctum and canaliculus, through to the lacrimal sac. They are then opened maximally providing two points of elevation. This maneuver allows for easier identification of the plane of dissection during lacrimal flap creation. It is especially useful with complex anatomy and for junior trainees in helping to orient the dissection. No tissue or instrument damage has been noted, and because many ophthalmic units have a vitreoretinal service, the forceps may be readily available at no extra cost. PMID- 17413648 TI - The variable response of women with menopausal hot flashes when treated with sertraline. AB - OBJECTIVE: To evaluate the variable response of women when treated with a selective serotonin reuptake inhibitor (sertraline) to decrease hot flashes. DESIGN: A double-blind, placebo-controlled, crossover trial was conducted in 102 women aged 40 to 65 years who were experiencing hot flashes and not taking any hormone therapy. The original purpose of the study was to evaluate the effectiveness of sertraline for the treatment of hot flashes. After 1 week of baseline hot flash data collection, study participants were randomized to receive placebo or active drug (sertraline 50 mg) for 4 weeks. This intervention was followed by a 1-week washout and crossover to the opposite treatment for 4 weeks. The number and severity of hot flashes were measured. RESULTS: One hundred two women were enrolled in the study, and 87 completed the study. The average response was a statistically significant but clinically modest reduction in hot flash frequency and hot flash index (frequency x severity). These data on the average response have been previously published. Although the average response was modest, some women responded vigorously, others modestly, and some women actually worsened. This is a post hoc analysis of those data. Percentage of change was divided into three categories of clinical response: women with a clinically significant reduction (>or=30%, n=27), women with a nonsignificant reduction (<30% to none, n=28), and women with an increase (1%-100%, n=32). A vigorous response to sertraline for the treatment of hot flashes was related to activity level, education, and menopausal status. CONCLUSIONS: Women have markedly variable responses when treated with antidepressants for their hot flashes. We have begun to describe the characteristics of those most likely to respond to treatment with a selective serotonin reuptake inhibitor. PMID- 17413649 TI - The cross-sectional relationship between fortnightly exercise and knee cartilage properties in healthy adult women in midlife. AB - OBJECTIVE: Exercise has been strongly promoted for the prevention of the major diseases of aging in women, such as cardiovascular disease and osteoporosis. However, conflicting data exist as to whether exercise benefits joint health and thus helps protect against degenerative processes such as knee osteoarthritis. The aim of this study was to examine the cross-sectional relationship between exercise and knee joint cartilage in women at midlife. DESIGN: A total of 176 women aged 40 to 67 years with no clinical knee osteoarthritis or history of significant knee injury were recruited from the community and had magnetic resonance imaging performed on their dominant knee to assess tibial cartilage volume and defects. The women completed a questionnaire to determine fortnightly exercise participation and frequency. RESULTS: Fortnightly participation in exercise at an intensity that caused tachypnea and an increased pulse rate for at least 20 minutes was positively associated with the volume of the medial tibial cartilage (P=0.02). There was a tendency for the frequency of exercise at this intensity to be positively associated with medial tibial cartilage volume (P=0.07). Exercise participation was not significantly associated with the presence of knee cartilage defects. CONCLUSIONS: Participation in exercise that causes tachypnea and an increased pulse rate for at least 20 minutes was associated with greater medial tibial cartilage volume, but not knee cartilage defects in non-healthcare-seeking women at midlife. More frequent exercise also tended to be associated with greater medial tibial cartilage volume. Although our findings need to be confirmed longitudinally, they provide further support for a beneficial effect of regular exercise on diseases related to aging. PMID- 17413651 TI - The neural basis of scene preferences. AB - What is the neural correlate of preference that governs our spontaneous selection of visual information? With a rapid, event-related functional magnetic resonance imaging design, we showed that the viewing of highly preferred compared to less preferred scenes (as assessed by participant ratings) was associated with greater blood-oxygen level dependent responses in the right parahippocampal cortex but not in the lateral occipital complex, ruling out feed forward and attentional effects. Highly preferred images also produced greater activation in the ventral striatum, suggesting that perceptual preference might engage the conventional reward system. These results are consistent with the hypothesis that high activity in the parahippocampal cortex, an area with a high density of cortical mu-opioid receptors, may be experienced as cognitively pleasurable. PMID- 17413652 TI - Zincergic innervation of medial prefrontal cortex by basolateral projection neurons. AB - The basolateral amygdaloid complex is a site of origin for zinc-containing pathways in the brain; it is also known for its massive innervation of the medial prefrontal cortex. The presence, and potential neuromodulatory role, of zinc within this fundamental corticolimbic circuit has not been described. For this study, basolateral neurons innervating the medial prefrontal cortex were retrogradely labeled with FluoroGold, and zinc-containing neurons were identified using autometallography to visualize zinc selenium precipitates. Upon quantification of single-labeled and double-labeled cells, 35% of basolateral neurons projecting to medial prefrontal cortex were found to also contain zinc. We conclude that zinc may act as a neuromodulator for a substantial proportion of basolateral-medial prefrontal cortical innervation, therefore implicating zinc in corticolimbic function as well as pathology. PMID- 17413653 TI - Early and automatic syntactic processing of person agreement. AB - Mismatch negativity, an index of automatic cerebral activity in response to novel stimuli, was used to determine the onset of morphosyntactic processing in French. Stimuli were four two-word sentences made up of a pronoun (first or second person) and a verb (first or second person). Verb forms differed only in the inflectional suffix, which made the sentences either syntactically correct or incorrect. The mismatch negativity response was found to be modulated by the grammaticality of the agreement relation at 50-140 ms after inflection onset, corroborating the previous finding that morphosyntactic processing occurs early and out of the focus of attention. The role of the pronoun-suffix association probabilities in determining the observed timing of morphosyntactic processing is discussed. PMID- 17413654 TI - Neural-tube-derived neuroepithelial stem cells: a new transplant resource for Parkinson's disease. AB - To assess the feasibility of using neuroepithelial stem cells as a transplant source for Parkinson's disease, neuroepithelial cells were harvested from the neural tube, cultured and stereotactically transplanted into the striatum of a rat model of Parkinson's disease. In culture, neuroepithelial cells generated abundant neurospheres and differentiated into both neurons and glia. After transplantation, tyrosine-hydroxylase-positive cells were detected in the graft. Furthermore, an apomorphine-induced rotation test showed that the implanted cells successfully promoted functional recovery in animals that underwent this transplantation procedure. Our results demonstrate that neuroepithelial cells may be a new source of donor material for Parkinson's disease. PMID- 17413655 TI - Theta activity attenuation correlates with avoidance learning progress in gerbils. AB - Changes of cortical activity were examined with electrocorticograms sampled from gerbils during learning. Animals were subjected to tone-conditioned avoidance training in a shuttle-box. Electrocorticograms were recorded from an electrode placed over medial prefrontal cortex. Temporal patterns of theta activity were analyzed across 180 successive trials. With the start of conditioning strong theta activity occurred in each trial in the phase immediately after hurdle crossing. With reliable occurrence of conditioned responses, that is at the stage of retrieval of the avoidance response, the theta activity became reduced. A negative correlation exists between the theta power and the development of learning progress. Theta reduction thus could reflect decreasing demands on information processing in the course of avoidance success monitoring across trials. PMID- 17413656 TI - Brain activation during human male ejaculation revisited. AB - In a prior [O]-H2O positron emission tomographic study we reported brain regions involved in human male ejaculation. Here, we used another, more recently acquired data set to evaluate the methodological approach of this previous study, and discovered that part of the reported activation pattern was not related to ejaculation. With a new analysis of these ejaculation data, we now demonstrate ejaculation-related activations in the deep cerebellar nuclei (dentate nucleus), anterior vermis, pons, and ventrolateral thalamus, and, most importantly, ejaculation-related deactivations throughout the prefrontal cortex. This revision offers a new and more accurate insight into the brain regions involved in human male ejaculation. PMID- 17413657 TI - Attenuated amyloid-beta aggregation and neurotoxicity owing to methionine oxidation. AB - Aggregation of the amyloid-beta (Abeta) peptide into amyloid plaques is a characteristic feature of Alzheimer's disease neuropathogenesis. We and others have previously demonstrated delayed Abeta aggregation as a consequence of oxidizing a single methionine residue at position 35 (Met-35). Here, we examined the consequences of Met-35 oxidation on the extremely aggregation-prone peptides Abeta1-42 and Abeta1-40Arctic with respect to protofibril and oligomer formation as well as neurotoxicity. Size exclusion chromatography and mass spectrometry demonstrated that monomer/dimers prevailed over larger oligomers after oxidizing Met-35, and consequently protofibril formation and aggregation of both Abeta1-42 and Abeta1-40Arctic were delayed. The oxidized peptides completely lacked neurotoxic effects in cortical neuronal cultures under these conditions, in contrast to the neurotoxic properties of the unoxidized peptides. We conclude that oxidation of Met-35 significantly attenuates aggregation of Abeta1-42 and Abeta1-40Arctic, and thereby reduces neurotoxicity. PMID- 17413658 TI - Functional motoneurons develop from human neural stem cell transplants in adult rats. AB - We have shown previously that primed human fetal neural stem cells, after transplantation into rat spinal cords, differentiated into cholinergic motoneurons that sent axons to contact medial gastrocnemius myocytes. Here we demonstrate that (i) axons from the transplanted cells are cholinergic and myelinated, (ii) putative synapses form on transplanted somata and dendrites in the ventral horn, (iii) human fetal neural stem cells transplantation led to normal electromyograms from medial gastrocnemius muscles, and (iv) the gait of transplanted animals was much improved. Accumulatively, our data indicate that some transplanted human fetal neural stem cells in adult motoneuron-deficient ventral horns differentiate into relatively normal motoneurons that are integrated into spinal and peripheral circuitry. These findings are steps towards the long-term goal of providing stem cell transplants for motoneuron loss. PMID- 17413659 TI - Wide spectrum modulation by KP-544 in models relevant for neuronal survival. AB - Reduced neurotrophic signalling has been proposed as a part of the pathophysiology behind neuronal death and dysfunction. The small molecule KP-544 was developed with the intention to enhance nerve growth factor signalling. To characterize the actions of KP-544 pharmacologically, we used four diverse models with relevance for neuronal function and survival. We found that 300-1000 nM KP 544 enhanced the neurite outgrowth in PC12 cells in response to a suboptimal concentration of nerve growth factor. KP-544 also protected the cerebellar granule cells from excitotoxicity apoptosis induced by the mitochondrial toxin methyl-phenyl-pyridinium, and modulated inflammation by inhibiting interleukin-6 production in primary astrocytes. Chronic treatment of rats with KP-544 prevented the hyper-responsiveness to amphetamine of animals treated with methylazoxymethanol acetate, a recently described neurodevelopmental model of schizophrenia. PMID- 17413660 TI - BoNT/E prevents seizure-induced activation of caspase 3 in the rat hippocampus. AB - Clinical and experimental studies clearly demonstrate that prolonged seizures and status epilepticus induce neuronal cell death in the brain. Recent evidence suggests that induction of apoptosis contributes greatly to seizure-induced brain damage. We recently demonstrated that intrahippocampal delivery of botulinum neurotoxin E (BoNT/E) in the rat hippocampus is able to prevent neuronal loss, which occurs after kainic-acid-induced seizures. Here, we investigated the molecular mechanisms of BoNT/E-mediated neuroprotection. We found that intrahippocampal administration of BoNT/E prevents the upregulation of apoptotic proteins (phosphorylated c-Jun and cleaved caspase 3), which occurs in hippocampal neurones following kainic acid seizures. These results demonstrate that the neuroprotective action of BoNT/E on seizure-injured hippocampal neurons involves the blockade of well-characterized apoptotic pathways. PMID- 17413661 TI - Increased KPI containing amyloid precursor protein in experimental autoimmune encephalomyelitis brains. AB - Amyloid precursor protein can be translated from three alternatively spliced mRNAs. We measured levels of amyloid precursor protein isoforms containing the Kunitz protease inhibitor domain (KPIAPP), and amyloid precursor protein without the Kunitz protease inhibitor domain (KPIAPP) in brain homogenates of acute experimental autoimmune encephalomyelitis mice. At the preclinical phase of the disease, both KPIAPP and KPIAPP levels were significantly higher in homogenates from brains of autoimmune encephalomyelitis mice, whereas at the acute phase of the disease only KPIAPP remained significantly elevated compared with controls. At the recovery phase, no differences were observed between the groups. The early and isoform-specific elevation of KPIAPP in autoimmune encephalomyelitis mice suggests a possible role for amyloid precursor protein in the immune response mediating the disease. PMID- 17413662 TI - Newborn neurons with hilar basal dendrites hallmark epileptogenic networks. AB - Basal dendrites are transiently present on developing newly generated granule cells in adult rats. In epileptic rats, basal dendrites often fail to retract, resulting in the generation of persisting hilar basal dendrites that have been proposed to contribute to hyperexcitability. Here, we demonstrate that the formation of hilar basal dendrites also occurs in the absence of seizures following induction of an epileptogenic neuronal network by amygdala kindling. The number of newly generated neurons with hilar basal dendrites was significantly increased 6-8 weeks after the last kindled seizure. Thus, even in periods without seizure activity, persistence of hilar basal dendrites presents a hallmark of hyperexcitable epileptogenic networks in the rodent brain and may contribute to disease progression. PMID- 17413663 TI - Central ghrelin induces feeding driven by energy needs not by reward. AB - Centrally administered ghrelin, the endogenous agonist of the growth hormone secretegogue receptor, powerfully stimulates food intake. Although the orexigenic action of this peptide has been well established, it remains unclear whether ghrelin-induced hyperphagia is driven by energy needs or by reward. In our study ghrelin was injected into the lateral cerebral ventricle or the hypothalamic paraventricular nucleus of rats given a choice between a palatable yet calorie dilute sucrose solution and a calorically dense chow. As a result of intraventricular and hypothalamic paraventricular ghrelin injections, animals increased intake of chow but not sucrose. When the sucrose solution was offered as the only source of calories, rats treated with ghrelin infused in the ventricle and site-specifically increased sucrose consumption. These results suggest that the primary effect of ghrelin is to stimulate food intake to satisfy energy needs. PMID- 17413664 TI - Na,K-ATPase generates calcium oscillations in hippocampal astrocytes. AB - Na,K-ATPase maintains not only ionic homeostasis, but also participates in a multiprotein complex mediating intracellular signalling. We show that ouabain, a specific ligand for Na,K-ATPase, evokes calcium oscillations in hippocampal astrocytes in primary cultures. Coimmunoprecipitation studies suggest that the mechanism underlying these calcium oscillations involves a multiprotein complex consisting of ankyrin-B, the inositol 1,4,5-trisphosphate receptor and Na,K ATPase. The ouabain/Na,K-ATPase multi-protein complex induced calcium-dependent downstream activation of the transcription factor nuclear factor-kappaB. Calcium oscillations and nuclear factor-kappaB activation were blocked following intracellular calcium store depletion. Thus, the specific Na,K-ATPase ligand ouabain induced inositol 1,4,5-trisphosphate receptor-dependent calcium oscillations in hippocampal astrocytes, which mediates nuclear factor-kappaB activation. PMID- 17413665 TI - The right-hemispheric auditory cortex in humans is sensitive to degraded speech sounds. AB - We investigated how degraded speech sounds activate the auditory cortices of the left and right hemisphere. To degrade the stimuli, we introduce uniform scalar quantization, a controlled and replicable manipulation, not used before, in cognitive neuroscience. Three Finnish vowels (/a/, /e/ and /u/) were used as stimuli for 10 participants in magnetoencephalography registrations. Compared with the original vowel sounds, the degraded sounds increased the amplitude of the right-hemispheric N1m without affecting the latency whereas the amplitude and latency of the N1m in the left hemisphere remained unaffected. Although the participants were able to identify the stimuli correctly, the increased degradation led to increased reaction times which correlated positively with the N1m amplitude. Thus, the auditory cortex of right hemisphere might be particularly involved in processing degraded speech and possibly compensates for the poor signal quality by increasing its activity. PMID- 17413666 TI - Activation of SI is modulated by attention: a random effects fMRI study using mechanical stimuli. AB - Animal experiments on tactile attention suggest a modulation of sensory processing on the level of sensory representations but correspondent neuroimaging data in humans is inconclusive. The present experiment used mechanical stimuli to study tactile processing while varying the focus of attention. Activations were contrasted between attend and ignore conditions, both of which employed identical stimulation characteristics and an active task. Random effects analysis revealed significant attention effects in area SI (primary somatosensory cortex) in that the blood oxygenation level-dependent response was greater for attended than for ignored stimuli. Modulations were further found in the secondary somatosensory cortex and the middle temporal gyrus. These findings suggest that stimulus processing at the level of primary representations in area SI is modulated by attention. PMID- 17413667 TI - Binaural processing in children using bilateral cochlear implants. AB - Binaural auditory brainstem processing was examined using evoked potential measures in 40 children who were implanted early and received a second implant simultaneously or after long or short periods of unilateral implant use. Wave latencies were shorter when evoked by the experienced versus naive implanted ear at initial bilateral activation. Binaural difference waves were detected in most children in response to apical but not basal electrode stimulation and were prolonged in latency in children implanted after long or short delays between implants. Timing differences between the implanted ears in children receiving sequential but not simultaneous bilateral implants reflect a relative immaturity of pathways innervating the second ear and results in abnormal timing of binaural processing at this initial implant stage. PMID- 17413668 TI - The mirror-neuron system: a Bayesian perspective. AB - Is it possible to understand the intentions of other people by simply observing their movements? Many neuroscientists believe that this ability depends on the brain's mirror-neuron system, which provides a direct link between action and observation. Precisely how intentions can be inferred through movement observation, however, has provoked much debate. One problem in inferring the cause of an observed action, is that the problem is ill-posed because identical movements can be made when performing different actions with different goals. Here we suggest that this problem is solved by the mirror-neuron system using predictive coding on the basis of a statistical approach known as empirical Bayesian inference. This means that the most likely cause of an observed movement can be inferred by minimizing the prediction error at all cortical levels that are engaged during movement observation. This account identifies a precise role for the mirror-neuron system in our ability to infer intentions from observed movement and outlines possible computational mechanisms. PMID- 17413669 TI - An important message from Medicare: new rules on July 1, 2007. PMID- 17413670 TI - Case managers and ethics: balancing advocacy with objective evaluation. PMID- 17413671 TI - Case managers optimize patient safety by facilitating effective care transitions. AB - In this new era of patient safety, the case manager, as an advocate and facilitator of care, has a pivotal role on the front line of healthcare delivery. Effective communication and collaboration between disciplines is key to the promotion of patient safety, and ultimately the avoidance of life-threatening medical errors. Across the healthcare continuum and within hospitals in particular, patients are routinely transferred from one service to another, from one level of care to another, or from one provider to another. As patients are stabilized and transitioned through the hospital system, there are multiple hand offs of care or care transitions that can often expose the patient to fragmented service and increase the risk of communication breakdown. Ineffective hand-offs can result in a disruption of continuity between one level of care and the next. In a culture that places a strong emphasis on patient safety, case managers can facilitate opportunities that ease care transitions whereby a change in venue is no longer perceived as a disruption in the flow of care but rather is viewed as a coordinated changeover where cautious and comprehensive communication sets the tone for the continued delivery of safe and effective healthcare. PMID- 17413673 TI - Designing a system of case management for a rural nursing clinic for elderly patients with depression. AB - This article provides an overview of the process and procedures used to develop and implement a system of case management for middle-aged and older depressed adults in a rural health nursing clinic. This system included on-site case management for elderly clients and telephonic follow-up by case managers on an ongoing basis. PMID- 17413675 TI - The work of nurse case managers in a cancer and cardiovascular disease risk screening program. AB - PURPOSE OF THE STUDY: The purpose of this conceptual model of nursing and health policy-based study was to identify the frequency and correlates of activities performed by nurse case managers. PRIMARY PRACTICE SETTINGS: Massachusetts Women's Health Network (WHN) contracting organization sites for breast and cervical cancer and cardiovascular disease risk screening. METHODOLOGY AND SAMPLE: Twenty nurse case managers were interviewed. RESULTS: More time was spent performing client service activities than bureaucratic activities. Frequently performed client service activities were tracking test results, finding/connecting with clients, assessing client needs, and educating clients. The most frequently performed activity was documenting services; the least, discharging clients. Client service activity frequency was correlated with client caseload size, social barriers, overall workload, satisfaction with the way activities are carried out in the WHN, special training in WHN policies and procedures, and contracting organization service delivery arrangements. Bureaucratic activity frequency was correlated with caseload size, workload, months as a WHN case manager, system barriers, satisfaction with the way activities were carried out in the WHN, and special training. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Documentation requires a great deal of WHN nurse case managers' time, which perhaps could be more productively spent with clients. Thus, more efficient ways to document services need to be identified. Additional research is needed to determine similarities and differences in activities performed by WHN nurse case managers and other case managers working in cancer and cardiovascular disease screening programs. Strategies need to be identified to remove all barriers that interfere with performance of case manager practice activities. Strategies are needed to reduce client fear of bills, overcome scheduling constraints, and improve translation services to lessen language barriers to effective communication. PMID- 17413676 TI - Anticoagulation in ambulatory care: an evidence-based review of the literature. AB - PURPOSE OF THE MANUSCRIPT: Anticoagulation is warranted for the treatment of various disorders including cardiac, vascular, and postsurgical causes. Many centers have nurse case managers to coordinate care for patients on anticoagulation. This increases the demand for specific guidelines to assist nurse case managers to ensure quality of care. This review will address guidelines for nurse case managers and providers regarding initiating anticoagulation treatment and monitoring prothrombin time and international normalized ratio. Information will also be provided regarding when the nurse case manager should notify the providers to establish target international normalized ratio. This review will also provide educational tools to serve as standards for patient teaching, including drug and food interactions. PRIMARY PRACTICE SETTING(S): This article applies to adult ambulatory practice that includes primary care, cardiology, and vascular and surgical settings. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: The complexity of managing anticoagulation in ambulatory practice warrants case management. The nurse case manager will establish a rapport with patients to improve compliance, providing patient education about diet, dosages, and drug interactions to reduce medication errors and bleeding complications. This review on anticoagulation management will assist nurse case managers and providers to provide better quality of care. PMID- 17413677 TI - Crucial conversations for healthcare: how to discuss lack of support, poor teamwork, and disrespect, Part II of III. PMID- 17413678 TI - Communication is the key for a successful return to work. PMID- 17413680 TI - Alphabet soup in case management. PMID- 17413681 TI - Effective workers' compensation nurse case management. PMID- 17413682 TI - Anogenital warts knowledge and counseling practices of US clinicians: results from a national survey. AB - OBJECTIVES: To examine messages US clinicians use when counseling patients diagnosed with anogenital warts. STUDY DESIGN: In mid-2004, we conducted a confidential mail survey of nationally representative samples of physicians practicing internal and adolescent medicine, family/general practice, obstetrics/gynecology, urology, or dermatology; nurse midwives; physician assistants; and nurse practitioners. The survey assessed knowledge and counseling practices of clinicians who had diagnosed anogenital warts. RESULTS: After adjusting for survey eligibility, 81% responded. Most (89%) were aware that human papillomavirus (HPV) causes anogenital warts, but only 48% were aware that oncogenic and wart-related HPV genotypes usually differ. Most (>95%) clinicians reported telling patients with warts that warts are an STD, are caused by a virus, or that their sex partners may have or may acquire warts. Many clinicians (>/=85%) also reported discussing STD prevention or assessing STD risk with such patients. Most reported addressing ways to prevent HPV (89%), including using condoms; limiting sex partners or practicing monogamy; or abstinence. Many also reported recommending prompt (82%) or more frequent (52%) Pap testing to female patients with anogenital warts. Potential barriers to counseling included providing definitive answers on how HPV infection was acquired, dealing with patients' psychosocial issues, and inadequate reimbursement. CONCLUSIONS: Most surveyed clinicians appropriately counseled patients about the cause and prevention of anogenital warts. However, many clinicians were unaware that oncogenic and wart-related HPV types usually differ, and this may explain why many reported recommending more aggressive cervical cancer screening for female patients with warts. PMID- 17413683 TI - Evaluation of a population-based program of expedited partner therapy for gonorrhea and chlamydial infection. AB - OBJECTIVE: To evaluate a partner notification program for gonorrhea and chlamydial infection that involves communitywide access to free patient-delivered partner therapy (PDPT) and use of case-report forms to triage patients to receive partner notification assistance. METHODS: We evaluated program components in randomly selected cases and compared outcomes before and after program institution. RESULTS: Following institution of the program, the percentage of cases who received PDPT from their diagnosing clinician increased from 5.6% to 16% (adjusted OR 3.2, 2.5-4.1). Among randomly selected cases, those referred to the health department via the case-report form were significantly more likely than nonreferred cases to have untreated sex partners (76% vs. 35%, OR 6.0, 95% CI 4.5-8.0), to accept PDPT from the health department (36% vs. 14%, 3.3, 95% CI 2.4-4.7), and to request that health department staff notify a partner for them (11% vs. 3%, OR 3.5, 95% CI 1.8-6.7). The percentage of cases classified as having all of their partners treated increased from 39% to 65% concurrent with institution of the program. CONCLUSIONS: A public health program that promotes routine use of PDPT and referral of selected patients for partner notification assistance appears to have improved partner notification outcomes. PMID- 17413684 TI - Efficacy and safety of etravirine (TMC125) in patients with highly resistant HIV 1: primary 24-week analysis. AB - OBJECTIVE: TMC125-C223 is an open-label, partially blinded, randomized clinical trial to evaluate the efficacy and safety of two dosages of etravirine (TMC125), a non-nucleoside reverse transcriptase inhibitor (NNRTI) with activity against wild-type and NNRTI-resistant HIV-1. DESIGN: A total of 199 patients were randomly assigned 2: 2: 1 to twice-daily etravirine 400 mg, 800 mg and control groups, respectively. The primary endpoint was a change in viral load from baseline at week 24 in the intention-to-treat population. METHODS: Patients had HIV-1 with genotypic resistance to approved NNRTIs and at least three primary protease inhibitor (PI) mutations. Etravirine groups received an optimized background of at least two approved antiretroviral agents [nucleoside reverse transcriptase inhibitors (NRTI) and/or lopinavir/ritonavir and/or enfuvirtide]. Control patients received optimized regimens of at least three antiretroviral agents (NRTIs or PIs and/or enfuvirtide). RESULTS: The mean change from baseline in HIV-1 RNA at week 24 was -1.04, -1.18 and -0.19 log10 copies/ml for etravirine 400 mg twice a day, 800 mg twice a day and the control group, respectively (P < 0.05 for both etravirine groups versus control). Etravirine showed no dose related effects on safety and tolerability. No consistent pattern of neuropsychiatric symptoms was observed. There were few hepatic adverse events, and rashes were predominantly early onset and mild to moderate in severity. CONCLUSION: Etravirine plus an optimized background significantly reduced HIV-1 RNA levels from baseline after 24 weeks in patients with substantial NNRTI and PI resistance, and demonstrated a favorable safety profile compared with control. PMID- 17413685 TI - Week 24 efficacy and safety of TMC114/ritonavir in treatment-experienced HIV patients. AB - BACKGROUND: Agents for the treatment of HIV-1-infected patients with resistance to current antiretroviral (ART) drugs are needed. METHODS: TMC114-C202 was a randomized, partially blinded, dose-finding study in treatment-experienced HIV-1 infected patients with one or more primary protease inhibitor (PI) mutations and HIV-1 RNA > 1000 copies/ml. Patients were randomized to receive one of four TMC114 doses given with ritonavir (TMC114/r) or investigator-selected control PI drug(s) (CPI); all received an optimized background regimen. The primary intent to-treat analysis compared the proportion of patients achieving a >or= 1 log10 copies/ml HIV-1 RNA reduction at week 24 between the treatment arms using the time-to-loss of virological response algorithm. RESULTS: For 278 patients at baseline, mean HIV-1 RNA was 4.7 log10 copies/ml, median CD4 cell count was 106 cells/mul; HIV-1 isolates had a median of three primary PI mutations and a median fold change in lopinavir susceptibility of 80. Discontinuation rates were 23% for TMC114/r versus 64% for CPI. More patients in each TMC114/r dose group achieved >or= 1.0 log10 copies/ml reduction in HIV-1 RNA than in the CPI group (45-62% versus 14%; P 400 copies/ml. METHODS: Accumulation of resistance between t0 and t1 was measured using genotypic susceptibility scores (GSS) obtained by counting the total number of active drugs (according to the Rega system v6.4.1) among all licensed antiretrovirals as of 1 January 2006. Patients were grouped according to the number of active drugs in the failing regimen at t0 (GSS_f-t0). RESULTS: At t0, patients had been on the failing combination antiretroviral therapy (cART) for a median of 11 months (range, 6-50 months). Even patients with extensive resistance to the failing regimen were still receiving benefit from treatment. An overall 6-monthly increase of 1.96 (SD, 2.23) International Aids Society-mutations and an average loss of 1.25 (SD, 1.81) active drugs were estimated. In comparison with patients with GSS_f-t0 = 0, the number of active drugs lost was -1.08 [95% confidence interval (CI), -2.13 to -0.03; P = 0.04] in those with GSS_f-t0 of 0.5-1.5 and -1.24 (95% CI, -2.44 to -0.04; P = 0.04) in those with GSS_f-t0 >or= 2. CONCLUSIONS: In patients kept on the same virologically failing cART regimen for a median of 6 months, there was considerable accumulation of drug resistance mutations, particularly in patients with initial low level of resistance to the failing regimen. Randomized comparisons of maintenance treatment strategies while awaiting a new suppressive therapy to become available are warranted. PMID- 17413694 TI - Nevirapine clearance from plasma in African adults stopping therapy: a pharmacokinetic substudy. AB - OBJECTIVE: To measure nevirapine elimination in African adults undertaking a structured treatment interruption (STI) in the DART trial. DESIGN: Cohort (16 women, 5 men; median weight 61 kg) within a randomized trial of management strategies. METHODS: Plasma nevirapine was measured by validated high performance liquid chromatography at 0,1,2,3 and 4 weeks after stopping the drug in a subset of patients undertaking an STI. All patients continued lamivudine plus zidovudine/stavudine for a further 7 days. RESULTS: Two patients with no or low plasma nevirapine concentration at baseline were excluded. Geometric mean plasma concentration when nevirapine was stopped in the remaining 19 patients was 6421 ng/ml (range, 3724-9473). Nevirapine was detected in 15/18 (83%) patients at 1 week, and 5/19 (26%) patients at 2 weeks but was not found any samples collected after 2 weeks. Only one patient had > 100 ng/ml (limit of quantification) at 2 weeks (415 ng/ml, female). The median times to reach thresholds of 200, 100 and 20 ng/ml (limit of detection) were estimated to be 7.6 [interquartile range (IQR), 7.0-10.1], 9.3 (IQR, 8.7-13.0) and 13.2 (IQR, 12.3-18.4) days, respectively, with 3/19 (16%) and 14/19 (74%) estimated to have reached < 20 ng/ml by 7 and 14 days, respectively. CONCLUSION: Although elimination of nevirapine was faster than previously published after a single dose, the data suggest that an additional staggered period of 7-10 days with dual nucleotide reverse transcriptase inhibitor cover is necessary for African patients discontinuing nevirapine. PMID- 17413695 TI - Prevalence and determinants of HIV infection in South India: a heterogeneous, rural epidemic. AB - OBJECTIVES: To assess the prevalence and determinants of HIV infection in the general population in Bagalkot district, a largely rural district in the southern Indian state of Karnataka. METHODS: Approximately 6700 individuals aged 15-49 years were randomly sampled from 10 villages and six towns, from three of Bagalkot's six sub-districts. Each consenting respondent was administered a questionnaire, followed by blood collection and testing for HIV, syphilis, and herpes simplex virus type 2 (HSV-2) on a 25% sub-sample. RESULTS: HIV prevalence was 2.9% overall, 2.4% in urban areas and 3.6% in rural areas [odds ratio (OR), 0.65; 95% confidence interval (CI), 0.45-0.95]. Significant differences in HIV prevalence were seen between the three sub-districts, with prevalences of 1.1, 3.0 and 6.4% (P < 0.05), and HIV prevalence in the 10 villages ranged from 0 to 8.2%. Reported multiple sexual partners, receiving money for sex and a history of medical injections were significantly associated with HIV infection, as were older age, being widowed, divorced, separated or deserted, lower education levels and being a woman of a lower caste. There was a strong association between HSV-2 and HIV infection (OR, 5.2; 95% CI, 2.3-11.5). CONCLUSIONS: The rural nature of this epidemic has important implications for prevention and care programs. The striking differentials observed in HIV prevalence between sub-districts and even villages suggest that risk and vulnerability for HIV are highly heterogeneous. Further research is required to understand the individual and community-level factors behind these differentials, so that preventive interventions can be directed to where they are most needed. PMID- 17413696 TI - Hormonal contraception and HIV-1 disease progression among postpartum Kenyan women. AB - OBJECTIVE: To assess the immediate and longer-term effects of the use of hormonal contraception on the progression of HIV-1 disease in postpartum women. DESIGN: A prospective cohort study. METHODS: Information on contraceptive use, breastfeeding and intercurrent illnesses was obtained from HIV-infected postpartum Kenyan women monthly in the first year postpartum and quarterly in the second year. Blood was collected for T-cell subset analyses and HIV-1-RNA levels at months 1, 3, 6, 9, 12, 18, and 24 postpartum. The immediate effect of the initiation of oral contraceptive pills (OCP) and depot medroxyprogesterone acetate (DMPA) was assessed by comparing the change in the HIV-1-RNA plasma viral load and CD4 T-cell counts among women remaining off these contraceptive methods with those initiating them. The longer-term effects of OCP and DMPA on disease progression were assessed using Loess curves and linear mixed effects models to compare changes over the first 24 months postpartum in these same disease progression markers. RESULTS: There were no significant immediate or longer-term effects of the use of OCP or DMPA on HIV-1-RNA plasma viral loads and CD4 T-cell counts in this cohort of HIV-infected postpartum Kenyan women. CONCLUSION: Comprehensive contraceptive counselling for HIV-1-infected women requires an understanding of the effects of various contraceptive methods on HIV-1 disease progression. In this study, hormonal contraception reassuringly had no immediate or longer-term effects on the rate of disease progression in chronically HIV-1 infected postpartum women. This highly effective family planning method may provide a useful and safe option for the prevention of mother-to-child transmission of HIV-1. PMID- 17413697 TI - Infectious correlates of HIV-1 shedding in the female upper and lower genital tracts. AB - OBJECTIVES: To determine the effects of vaginal, cervical, and endometrial infections on shedding of HIV-1 RNA in the female genital tract. DESIGN: Cross sectional. METHODS: Antiretroviral-naive women from Nairobi, Kenya with CD4 cell counts >or= 350 cells/mul had plasma and endocervical wick samples collected for HIV quantification by real-time RNA reverse transcriptase-polymerase chain reaction. Vaginal and cervical Gram stains and endometrial biopsies were obtained. Vaginal Gram stain was used to diagnose bacterial vaginosis and to quantify Lactobacillus levels. RESULTS: Twenty-six of 50 (52%) women had detectable endocervical HIV-1 RNA with a median endocervical viral load of 1760 copies/ml (range: undetectable to 1 1,030,000 copies/ml). Women with decreased Lactobacillus had 15.8-fold [95% confidence interval (CI), 2.0-123] greater endocervical HIV-1 RNA than women with normal Lactobacillus levels. Women with plasma cell (PC) endometritis [>or= 1 PC/high-power field (hpf)] had a 15.8-fold (95% CI, 2.0-120) higher endocervical HIV RNA level than women without PC endometritis. Both these associations remained after controlling for plasma viral load. Cervicitis (>or= 30 polymorphonuclear leukocytes/hpf), however, was not associated with endocervical HIV-1 RNA shedding (P = 0.81). CONCLUSIONS: In HIV-1 infected, antiretroviral-naive women without symptoms of pelvic inflammatory disease infection, abnormal vaginal flora and inflammatory cells in the endometrium affected HIV-1 shedding from the lower genital tract. These data suggest that both the upper and lower genital tracts contribute to female HIV-1 genital shedding. PMID- 17413698 TI - Long-term follow-up of patients taking tenofovir DF with low-level HIV-1 viremia and the K65R substitution in HIV-1 RT. AB - Patients with on-going HIV-1 replication and a K65R mutation in HIV-1 RT were assessed for further development of RT mutations while taking tenofovir disoproxil fumarate and other antiretroviral drugs. K65R was observed in 10 out of 536 treatment-experienced patients entering the study. K65R became undetectable in two patients, and the development of additional resistance mutations was minimal. Over 18 months, no patient developed multinucleoside resistance (Q151M or T69 insertions) and plasma viral loads were stable (median +0.04 log10 copies/ml). PMID- 17413699 TI - Antimalarial activity of sera from subjects taking HIV protease inhibitors. AB - Synergy between HIV and malaria is being increasingly recognized. We examined the antimalarial activity of sera from subjects receiving chloroquine, no drugs or HAART. Sera from subjects taking ritonavir-boosted saquinavir or lopinavir significantly inhibited parasite growth (median of 55 and 69% inhibition, respectively). These results indicate that patients on protease inhibitors may be afforded some protection from malaria. The clinical relevance of these observations will require confirmation in controlled studies in malaria-endemic regions. PMID- 17413700 TI - Nearly identical strains of human herpesvirus 8 in couples discordant for Kaposi's sarcoma. AB - Transmission of human herpesvirus 8 (HHV-8) may occur through various routes including breastfeeding and sexual intercourse. We attempted to detect HHV-8 infection in nine HIV-positive couples discordant for Kaposi's sarcoma who maintained a monogamous sexual relationship for at least one year. By quantitative real-time polymerase chain reaction and HHV-8 genotyping we provide strong evidence for the sexual transmission of HHV-8 in this unique cohort. PMID- 17413701 TI - Long-term safety, effectiveness and quality of a generic fixed-dose combination of nevirapine, stavudine and lamivudine. AB - We assessed the long-term safety, effectiveness and quality of a fixed-dose combination of nevirapine, stavudine and lamivudine (triomune). HIV-1-infected adults initially enrolled in a one-year, open-label, single-arm, multicentre trial in Cameroon were followed for 2 years. Our results support the safety and effectiveness of the triomune combination for first-line treatment of HIV infection. Virological effectiveness appeared to wane somewhat during the second year of treatment, however, and plasma nevirapine concentrations were relatively high. PMID- 17413702 TI - Hepatitis B reverse seroconversion in HIV-positive patients: case series and review of the literature. AB - Hepatitis B (HBV) reverse seroconversion is rare in HIV disease but can be fatal. We present a case series of 6 patients with reverse seroconversion and review 18 additional cases described in the literature. Elevated transaminases were seen in 13/21 (62%). Reverse seroconversion occurred more frequently in the setting of HIV virologic failure. Only 3 patients demonstrated reverse seroconversion in the setting of lamivudine or tenofovir withdrawal. 2/24 (8%) patients died from their HBV flare. PMID- 17413703 TI - Suspected immune reconstitution inflammatory syndrome associated with the proliferation of Kaposi's sarcoma during HAART. PMID- 17413704 TI - Failure of bone marrow transplantation to eradicate HIV reservoir despite efficient HAART. PMID- 17413705 TI - Response to Schmutz et al., 'Combination of tenofovir and lamivudine versus tenofovir after lamivudine failure for therapy of hepatitis B in HIV coinfection'. PMID- 17413706 TI - Intravenous ganciclovir consistently induces remission of persistent Epstein-Barr encephalitis in an HIV-1-infected patient. PMID- 17413707 TI - Posterior reversible encephalopathy syndrome in HIV patients: case report and review of the literature. PMID- 17413708 TI - Response to Blaas et al., 'Acute renal failure in HIV patients with liver cirrhosis receiving tenofovir: a report of two cases'. PMID- 17413709 TI - Acquired factor VIII inhibitor in an HIV-infected patient after treatment with pegylated interferon-alpha 2a and ribavirin. PMID- 17413710 TI - Interpreting the association between HIV and bone mineral density. PMID- 17413711 TI - Where's your prescription for living? PMID- 17413712 TI - Going forward. PMID- 17413713 TI - Condition H. PMID- 17413716 TI - Postpartum hemorrhage. PMID- 17413725 TI - Pain assessment in preterm neonates. PMID- 17413726 TI - Breastfeeding may reduce neonatal procedural pain. PMID- 17413727 TI - Stopping verbal abuse in the workplace. PMID- 17413728 TI - Recognizing quiet delirium. PMID- 17413731 TI - Emergency: pediatric orthopnea and total airway obstruction. PMID- 17413732 TI - Wound wise: PUPPI: the Pressure Ulcer Prevention Protocol Interventions. PMID- 17413735 TI - Tattoos, body piercing, and nursing: a photo essay. Interview by Jason P Smith. PMID- 17413736 TI - A brief overview of body art. PMID- 17413737 TI - The cancer survivor's prescription for living. PMID- 17413752 TI - Covering America's uninsured. PMID- 17413754 TI - Labor of love: nurse midwife Ruth Watson Lubic. Interview by Leslie Knowlton. PMID- 17413755 TI - Reflections: waiting for Kimberly. PMID- 17413756 TI - A systems biology approach to the diagnosis of venous thrombosis risk. PMID- 17413757 TI - Altered regulation of in-vivo coagulation in orthopedic patients prior to knee or hip replacement surgery. AB - Up to 20% of patients develop venographically proven deep-vein thrombosis after elective orthopedic surgery even under the cover of heparin or low molecular weight heparin. The extent to which the chronic inflammation of osteoarthritis requiring elective orthopedic surgery alters in-vivo coagulation and whether any specific alteration influences the development of postoperative thrombosis are unknown. This study compared the concentrations of activated factor VII (FVIIa), tissue factor pathway inhibitor (TFPI), activated factor X (FXa)-TFPI, thrombin antithrombin, and prothrombin fragment 1+2 (F1+2) in plasmas of 535 healthy individuals (ages 17-76) with those in the preoperative plasmas of 306 arthritis patients (ages 30-92) scheduled for elective knee or hip replacement surgery. C reactive protein was also measured in the plasmas of approximately 15% of the participants. Age-adjusted concentrations of FVIIa, F1+2, and C-reactive protein were higher in patients than controls, while the concentrations of thrombin antithrombin, TFPI and FXa-TFPI were similar. Chronic inflammation in the patients was thus associated with increased coagulation in vivo. Without compensatory increases in the concentrations of TFPI (natural inhibitor of prothrombinase), the elevated concentrations of FVIIa in the preoperative plasmas and the trauma associated with surgery may enhance the risk for developing postoperative deep-vein thrombosis. PMID- 17413758 TI - The effects of glycosaminoglycans on coagulation: a thromboelastographic study. AB - Endogenous heparinoids impair coagulation, evidenced by thrombelastography in cirrhotic patients with bacterial infection, but it is not clear which glycosaminoglycans can be detected by native and heparinase-modified thrombelastography. To assess the effects of different glycosaminoglycans on thrombelastography parameters and the reversibility of these effects by heparinase-I-modified thrombelastography. Twenty volunteers were enrolled. Solutions of heparan sulphate, dermatan sulphate, and chondroitin-4-sulphate were prepared at 'equivalent' concentrations, based on the composition and anticoagulant activity of danaparoid. Serial dilutions of each glycosaminoglycan were prepared to achieve 1.0, 0.5, 0.1, and 0.05 U/ml. Native and heparinase modified thrombelastography, anti-activated factor X activity and heparin cofactor II activity were evaluated at each concentration. A statistically significant heparin-like effect was seen with 1 and 0.5 U/ml heparan sulphate, and 1 and 0.5 U/ml dermatan sulphate, which was completely reversed by heparinase modified thrombelastography. Anti-activated factor X activity was significantly increased in samples containing heparan and dermatan sulphates. The heparin cofactor II activity decreased with 1.0 and 0.5 U/ml dermatan sulphate and chondroitin-4-sulphate, but not with heparan sulphate. Heparan and dermatan sulphates affect haemostasis when added to whole blood in vitro, detectable by native thrombelastography and completely reversed by heparinase-I-modified thrombelastography. They may therefore be responsible for the heparin-like effect seen by thrombelastography in patients with cirrhosis and bacterial infection. PMID- 17413759 TI - Antithrombin III metabolism in the pulmonary vessel endothelium. AB - In 85 patients undergoing aorto-coronary bypass for atherosclerotic coronary disease, we measured the antithrombin III activity levels and the thrombin antithrombin III complex concentrations in blood from the pulmonary and the radial arteries, taken before the aorto-coronary bypass procedure, with the aim of investigating the role of the pulmonary endothelium in the metabolism of the inhibitor. Results showed significantly lower mean antithrombin III activity levels, expressed as a percentage of normal plasma, in blood from the radial artery with respect to levels from the pulmonary artery (0.78 +/- 0.12 versus 0.80 +/- 0.12, P<0.0001), while no significant difference was found in thrombin antithrombin III complex concentrations. The results seem to show that the pulmonary endothelium contributes to the antithrombin III metabolism with a 0.023 breakdown rate, corresponding to about a 0.1 fraction of the reported 0.22-0.25 total body catabolic rate, as well as the pulmonary endothelial surface (50-70 m2) corresponding to about a 0.1 fraction of the peripheral vessels' endothelial surface (500-700 m2). The data support the hypothesis of a main endothelial catabolism of antithrombin III. PMID- 17413760 TI - Blood inhibitory capacity toward exogenous plasmin. AB - Stabilized, active plasmin is a novel thrombolytic for direct delivery to clots. Although it is known that protease inhibitors in plasma inhibit plasmin, the amount of plasmin that can be added to plasma/blood before free plasmin is observed is not clear. Determination of free plasmin activity in plasma using chromogenic substrates represents a challenge due to false-positive signals from plasmin entrapped by alpha2-macroglobulin. Size-exclusion chromatography was used to separate the plasmin-alpha2-macroglobulin complex from uninhibited, free plasmin. In this in-vitro study, exogenous plasmin is effectively inhibited up to 2.4 micromol/l after 5-min incubation with plasma at 37 degrees C. Initially, plasmin was consumed predominantly by alpha2-antiplasmin up to 1.2 micromol/l plasmin. Following exhaustion of alpha2-antiplasmin, plasmin was further consumed by alpha2-macroglobulin up to 2.4 micromol/l plasmin added to human plasma. Whole human blood was found to have an increased inhibitory capacity over that of plasma; free plasmin activity could be measured only above 3.8 micromol/l added plasmin. In conclusion, several mechanisms exist that control plasmin activity in human blood; in addition to alpha2-antiplasmin and alpha2-macroglobulin, blood cells contribute to the inhibition of exogenously administered plasmin. These in vitro results indicate that doses of plasmin up to approximately 12 mg/kg in humans can be completely inactivated by blood. PMID- 17413761 TI - A comparison of the Thrombelastograph and the ROTEM. AB - Elastic modulus-based assessment of hemostasis in clinical and research settings has been conducted for nearly 60 years. Two systems utilizing this technology include the Thrombelastograph (Haemoscope Corporation, Niles, Illinois, USA) and the ROTEM (Pentapharm GmbH, Munich, Germany). The study goal was to compare the Thrombelastograph and the ROTEM to determine whether differences in data could be detected. Plasma (n = 8 per condition) was not activated (native); was celite activated, hypocoagulable by dilution with hydroxyethyl starch; was celite activated, normal; and was celite-activated, hypercoagulable by addition of fibrinogen. Equivalent ratios of plasma mixture to CaCl2 were maintained between the Thrombelastograph and ROTEM systems. Data collected included the reaction time, angle, maximum amplitude and maximum elastic modulus. The ROTEM system generated significantly (P<0.05) smaller reaction time (mean +/- SD, 459 +/- 39 versus 788 +/- 94 s) and greater angle (60.1 +/- 5.3 versus 46.4 +/- 4.6 degrees ), maximum amplitude (27.6 +/- 1.6 versus 24.4 +/- 1.4 mm) and maximum elastic modulus (1911 +/- 152 versus 1616 +/- 121 dynes/cm2) values with native plasma compared with the Thrombelastograph. Celite-activation tended to attenuate differences between the two systems. In conclusion, a comparison of the Thrombelastograph and the ROTEM with similar samples demonstrated clinically significant differences in the data generated without exogenous activation. Given these data and the differences in proprietary activator, divergent results are expected, perhaps affecting clinical decision-making. PMID- 17413762 TI - Diagnostic value of D-dimer in outpatients with suspected deep venous thrombosis receiving oral anticoagulation. AB - D-dimer has proved a useful diagnostic tool for the exclusion of deep venous thrombosis (DVT). The objective of this paper was to evaluate the diagnostic performance of a diagnostic algorithm combining clinical probability and D-dimer in outpatients receiving oral anticoagulant treatment (OAT) similar to those regularly applied to nonanticoagulated individuals. We enrolled 70 outpatients on OAT who presented with clinically suspected DVT; a standard diagnostic algorithm including clinical evaluation using the modified Wells score and a quantitative immunoturbidimetric D-dimer assay (STA Liatest D-Di; Diagnostica Stago, Asnieres sur Seine, France) was used. A 3-month follow-up period was applied for those patients in whom DVT was initially excluded. The prevalence of DVT was 18.5% (13/70); four of the diagnoses were made during the 3-month follow-up period. The sensitivity, specificity and negative predictive value of D-dimer were 69.2% (95 confidence interval, 42.4-87.3), 47.4% (95% confidence interval, 35.0-60.1) and 87.1% (95% confidence interval, 71.1-94.9), respectively. In conclusion, D-dimer is of limited value in outpatients on OAT presenting with clinically suspected DVT and should be omitted in such individuals; these patients should always undergo compression venous ultrasound, and repeat ultrasonography within 1 week might be warranted in cases with an initial negative examination. PMID- 17413763 TI - Venous and arterial thrombosis in patients with HIV infection. AB - Thromboembolic complications in HIV-infected patients have been reported. To our knowledge, no case-control studies have compared the prevalence of thromboembolic events between HIV-positive and HIV-negative individuals. One hundred and sixty nine HIV-infected patients and 180 randomly selected blood donors were enrolled. Selected patients completed a specific questionnaire and were subsequently interviewed. Information was collected on family and personal history of cardiovascular disorders and the presence of personal risk factors for venous and arterial thrombosis. All reported events were adjudicated only if adequate documentation of objective tests was available. Mean age and sex were similar in the two groups. A vascular event was documented in six HIV-infected patients (3.55%) and in none of the controls (P=0.0108). Family history of cardiovascular disorders, cigarette smoking and hypertriglyceridemia were more prevalent in HIV patients than in controls. In multivariate analysis, neither family traditional cardiovascular risk factors nor HIV infection were independently associated with the presence of thromboembolic events. The results confirm the hypothesis that HIV-positive patients have an increased risk of thromboembolic disorders. Whether this increased risk has been provoked by HIV infection itself or by other associated risk factors for cardiovascular events, such as cigarette smoking and hypertriglyceridemia, remain to be clarified. PMID- 17413764 TI - The efficiency of anti-activated factor X and anti-activated factor II anticoagulants. AB - Six thrombin-generation inhibitors or thrombin inhibitors were compared in the extrinsic coagulation activity assay (EXCA), where the normal thrombin generation is about 1 IU/ml within 1 min (37 degrees C). Unfrozen pooled normal citrated plasma was supplemented on flat-bottom wells (23 degrees C) with increasing concentrations of dalteparin, danaparoid, heparin, fondaparinux, hirudin, or argatroban. To 50 microl plasma, 5 microl of 1.5 ng/ml tissue factor, 6% bovine serum albumin, and 250 mmol/l CaCl2 were added. After 1 and 2 min coagulation reaction time at 37 degrees C (EXCA-1 and EXCA-2), 100 microl of 2.5 mol/l arginine and 0.16% Triton X 100, pH 8.6, were added. After 3 min (23 degrees C), 25 microl of 1 mmol/l CHG-Ala-Arg-pNA in 1.25 mol/l arginine, pH 8.7, were added, and the linear increase in absorbance with time was determined at 405 nm. The 50% inhibitory concentrations of plasmatic anticoagulants tested in the EXCA-1 (37 degrees C) were 0.025 IU/ml dalteparin, 0.13 U/ml danaparoid, 0.12 IU/ml heparin, 1.3 microg/ml fondaparinux, 2.4 ng/ml hirudin, and 1 microg/ml argatroban. From the 50% inhibitory concentration of hirudin it can be concluded that inhibition of about 30 mIU/ml thrombin halves the normal EXCA-1 value (i.e. if about 0.1 IU/ml thrombin are inactivated, then thrombin cannot self-amplify its generation 10-fold). The efficiency of any clinically used plasmatic anticoagulant can be monitored in the EXCA. PMID- 17413765 TI - Haemostatic properties of human plasma subjected to a sterilizing dose of gamma irradiation in the presence of ascorbate. AB - The objective was to study the effects of gamma irradiation, in the presence of sodium ascorbate, on coagulation/fibrinolytic activity of fresh frozen plasma to be applied to inactivate the transfusion-transmitted viruses in plasma-derived products. Plasma was irradiated (50 kGy total dose, on dry ice) using a 60Co source. The plasma proteins were analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis and western blot and the following parameters estimated: prothrombin time, functional fibrinogen concentration, thrombin induced fibrinogen polymerization, plasminogen activity, and tissue-type plasminogen activator-induced conversion of plasminogen to plasmin. In irradiated plasma a moderate fragmentation of the most labile plasma proteins was found. The prothrombin time was prolonged (1.5-fold), functional fibrinogen was significantly reduced (60%), fibrinogen polymerization was impaired, plasminogen was predominantly maintained (90%) and tissue-type plasminogen activator-induced conversion of plasminogen to plasmin was unchanged. Ascorbate (25 mmol/l) raised the level of functional fibrinogen in irradiated plasma (to 50%; P=0.0245) and slightly accelerated its polymerization. The small protective effect of ascorbate might be due to inhibition of the radiation-induced fibrinogen oxidation and/or fragmentation but addition of other antioxidants/stabilizers would be crucial when a high irradiation dose, an effective treatment for inactivation of the most resistant viruses, is applied. PMID- 17413766 TI - Heparin-like anticoagulant associated with multiple myeloma and neutralized with protamine sulfate. AB - A 55-year-old man with multiple myeloma developed sustained bleeding after bone marrow aspiration and cutaneous bleeding. Routine coagulation studies revealed a prolonged activated partial thromboplastin time and thrombin time (> 60 s) with a normal reptilase time. Further evaluation showed failure of the activated partial thromboplastin time to correct completely in a 1: 1 mixture with normal plasma. Treatment of the patient's plasma in vitro with protamine sulfate normalized the thrombin time. The presence of a heparin-like anticoagulant was suspected. The plasma heparin level was 0.73 IU/ml. Intravenous infusion of protamine sulfate appeared to neutralize the anticoagulant activity and stop the bleeding. The cancer cells themselves or the invasive nature of this type of cancer might result in a massive release of a heparinoid. Such coagulopathy appears to be a rare mechanism of bleeding and it is an important entity to consider since it is potentially reversible with protamine sulfate. PMID- 17413767 TI - Severe prekallikrein deficiencies due to homozygous C529Y mutations. AB - Two consecutive severe prekallikrein deficiencies were investigated. The first was identified in a 63-year-old patient admitted for ischemic stroke. The second deficiency was identified in a 38-year-old patient admitted for a second trimester pregnancy loss. A homozygous C529Y mutation was identified for both cases, whereas they are unrelated and no consanguineous marriage is known from the patients. These data point to a possible high frequency of this mutation as a cause of prekallikrein deficiency. PMID- 17413768 TI - Clinically significant differences between point-of-care analysers and a standard analyser for monitoring the International Normalized Ratio in oral anticoagulant therapy: a multi-instrument evaluation in a hospital outpatient setting. AB - The increasing number of patients requiring oral anticoagulant therapy has lead to an expansion in the use of point-of-care test (POCT) analysers for measuring the International Normalized Ratio (INR) for monitoring purposes. Availability of new technology inevitably leads to comparisons with standard methodologies, and studies to date have reached varying conclusions about the comparability of POCT INRs with conventional testing. We compared the performance of five POCT instruments (Hemochron Junior Signature, ProTime, CoaguChek S, INRatio and TAS) against Innovin thromboplastin on a Sysmex CA1500 automated analyser. The Hemochron Junior Signature, ProTime and CoaguChek S demonstrated strong correlation with the laboratory method (R2 > 0.94). These three analysers demonstrated higher percentages of paired results within 0.5 INR units (81.5, 92.0 and 74.0%, respectively); the INRatio and TAS demonstrated 54.2 and 62.2%, respectively. Within INR ranges of up to 2.0, 2.1-3.0, 3.1-4.0 and above 4.0, none of the POCT analysers demonstrated significant agreement with the standard method in every range. All POCT instruments showed a degree of bias and greater variation from the standard method at INR values above 3.0. These data indicate the potential for POCT analysers to generate INR values sufficiently different from conventional methods that they may impact on clinical decision-making. PMID- 17413769 TI - Plasma S/R ratio of warfarin co-varies with VKORC1 haplotype. AB - We recently reported that the low-dose VKORC1*2 haplotype is an important genetic determinant for warfarin dose requirement and is associated with difficulties to attain stable therapeutic prothrombin time--International Normalized Ratio in patients undergoing anticoagulation therapy. The aim of this study was to investigate whether patients with VKORC1*2 compared with patients carrying high dose haplotypes VKORC1*3 or VKORC1*4 had different warfarin S/R ratios in their plasma, and whether that was related to CYP2C9 variants CYP2C9*2 and CYP2C9*3 or other factors. Samples from patients previously haplotyped for VKORC1 and measured for plasma warfarin concentration were genotyped for the CYP2C9 variants CYP2C9*2 and CYP2C9*3. Nonparametric statistical analysis was performed to elucidate whether there was any significant difference in the warfarin S/R ratio between the two patient groups. Our result shows that there is a significant difference (P<0.01) in warfarin S/R ratios between VKORC1*2 and VKORC1*3 or VKORC1*4 patients. This difference did not originate from CYP2C9 variants CYP2C9*2 and CYP2C9*3. We speculate that VKORC1 haplotypes possibly are linked to some unidentified factors involved in the metabolic clearance of warfarin enantiomers. Dose-dependent variations in (S)-warfarin and (R)-warfarin clearance in these patients can also be a probable explanation for the difference in warfarin S/R ratios. PMID- 17413770 TI - Perceptions of dyspnea and helpful interventions during the advanced stage of lung cancer: Chinese patients' perspectives. AB - Dyspnea is a distressful but neglected symptom in oncology practice and research. The aim of this study was to describe the experience of dyspnea and helpful interventions in Chinese patients with advanced lung cancer admitted in the palliative care unit in 1 region in Hong Kong. A qualitative description approach using in-depth interviews was used to guide this study. Eleven participants agreed to be interviewed with age ranging from 51 to 80 years. They have been diagnosed with lung cancer from 1 to 12 months, and all required oxygen therapy from dyspnea. The results of content analysis revealed 4 main themes: (1) characteristics of dyspnea, (2) impact of dyspnea, (3) strategies used to manage dyspnea, and (4) nurses' role in managing dyspnea. Patients in this study found no Chinese words to adequately define and describe dyspnea and relied on sensations they experienced during the dyspnea episode. The impact of dyspnea was multidimensional, and patients used various strategies to manage dyspnea, including avoiding triggers and utilizing traditional Chinese medicine. Healthcare professionals were perceived to play a very inadequate role in assisting patients with dyspnea, and participants suggested that they should take a more active role in educating and supporting patients with dyspnea. PMID- 17413771 TI - Knowledge and attitudes about cancer pain management: a national survey of Italian hospice nurses. AB - To obtain information about the knowledge and attitudes of Italian hospice nurses concerning cancer pain management and to determine the predictor of nurses' pain management knowledge. Nationwide descriptive study. Hospice nurses in Italy from 9 hospice units distributed in the north, center, and south of Italy. Sixty-six nurses completed the questionnaire, indicating a 66.6% response rate. The Nurses' Knowledge and Attitudes Survey (Italian version) and a background information form were used to collect the data. Knowledge and attitudes regarding cancer pain. Among the 39 pain knowledge questions assessed, the mean number of correctly answered question was 24.4 (SD = 4.2), with a range of 15 to 35 items correctly answered. The correct answer rate for the entire scale, on average, was 62.7% (SD = 28%). Further analysis of items showed that more than 30% of hospice nurses underestimated the patients' pain and they did not treat the pain in the correct way; they had an incorrect self-evaluation about their pain management knowledge. Results from stepwise regression showed that nurses with higher mean correct answer scores had attended more courses on pain education. From these results, we conclude that there are still significant knowledge deficits and erroneous beliefs that may hamper treatment of hospice patients in pain. The results of this study could be useful to institutions involved in the education and application of patient pain management. PMID- 17413772 TI - Reluctance to care: a systematic review and development of a conceptual framework. AB - Primary lay carers are increasingly important in the care of patients with cancer, but their role can be complex and extended. Potential carers may feel anything from highly committed to not at all interested in caregiving, but powerful social norms pressure them to accept the role, and reluctance may be hidden to avoid censure. The purpose of this review was to gain insights into caregiving reluctance and its consequences. The findings were organized into 4 major dimensions: demographic, physical, psychological, and social. Three major outcomes were identified: deterioration in the carer-patient relationship, reduced quality of care, and institutionalization. Definitive answers to the review questions remain elusive. Choice seems to be a major indicator of caregiving reluctance, although reluctance may not remain static over the caregiving trajectory. Caregiving reluctance remains an underexplored topic, particularly in the context of cancer. PMID- 17413773 TI - Younger women's perceptions of coping with breast cancer. AB - Numerous studies have demonstrated an association between coping strategies and better quality of life after breast cancer. Because younger women consistently show greater psychological morbidity than older women after breast cancer diagnosis, there is great interest in the coping strategies of younger women. The present cross-sectional study used quantitative and qualitative methods to examine coping strategies used by 201 women who were aged 50 years or younger at diagnosis and were 6 months to 3.5 years postdiagnosis. Quantitative results from a modified version of the Ways of Coping scale revealed that the most frequently used coping strategies were positive cognitive restructuring, wishful thinking, and making changes. Qualitative analyses based on open-ended questioning of how women best coped with different stressful aspects of their diagnosis showed that women reported finding different strategies useful depending on the stressor. For example, social support was helpful in dealing with anger or depression, whereas positive cognitive restructuring was more helpful for concerns about the future. Analyses also confirmed that most coping strategies cited in commonly administered coping scales were used frequently by these women. However, several coping strategies not generally measured were also deemed valuable, including engaging in physical activity, using medications, and resting. These findings suggest that clinicians should identify patients' particular stressors and help with coping techniques targeting particular concerns. PMID- 17413774 TI - Resting energy expenditure and body mass changes in women during adjuvant chemotherapy for breast cancer. AB - Weight gain is a commonly reported side effect of adjuvant chemotherapy. A change in resting energy expenditure during treatment has been a suggested mechanism for weight gain. We prospectively measured resting energy expenditure, weight change, and body composition (dual-energy x-ray absorptiometry) in 10 women undergoing adjuvant chemotherapy for breast cancer. There was no change in resting energy expenditure across cycles of chemotherapy (P =.78) or from baseline to the end of treatment (1,189.68 +/- 80.27 vs 1,205.76 +/- 56.71 kcal/d; P =.74). Overall, participants did not gain weight across treatment. However, there was an overall trend toward weight gain (66.3 +/- 5.1 vs 68.2 +/- 5.0 kg; P =.09), and participants did show an increase in total fat mass (24.2 +/- 3.8 vs 26.5 +/- 3.2 kg; P =.04), whereas muscle mass remained the same. Although no change in resting energy expenditure was seen, the observed increase in total fat mass is consistent with a decrease in physical activity level commonly reported with adjuvant chemotherapy treatment of breast cancer, and these body composition changes may have important health implications for survivors. PMID- 17413775 TI - Stressors and coping strategies among female cancer survivors after treatments. AB - Although cancer survivors often face stressors and experience psychologic symptoms and distress, research on the particular stressors and coping strategies upon finishing cancer treatments is rare. The study purposes were to identify the stressors experienced and the strategies used by women cancer survivors at this phase of survivorship. The specific aims were to describe the stressors at the end of primary cancer treatments and delineate coping strategies that were used and helpful. Using a longitudinal design, interviews were conducted within 4 weeks and 3 to 4 months after treatment. Participants (N = 51) were women aged 34 to 77, and had radiation and/or chemotherapy for primary breast or gynecologic cancers. Participants' primary stressors at the end of treatment included feelings of uncertainty about treatment, follow-up, and symptoms; physical concerns; difficulty concentrating, attitudes about body, and dealing with mortality. Participants used acceptance, religion, and distraction as primary coping strategies. These strategies also were rated highly as helpful coping strategies. Clinicians can provide anticipatory guidance, based upon previous survivors' strategies, as well as assess and address particular stressors at this phase. Nurses can design and test patient-centered interventions that address women's specific stressors and promote effective coping strategies among women at this phase of survivorship. PMID- 17413776 TI - Facilitating oral chemotherapy treatment and compliance through patient/family focused education. AB - Oral chemotherapy is assuming an increasingly important role in cancer therapy. Pharmaceutical firms continue to invest heavily in oral drug development with approximately 25% of more than 400 antineoplastic drugs currently in the development pipeline planned as oral agents. New treatments, patient preference, and the economic realities of cancer care delivery present physicians, nurses, pharmacists, and cancer center administrators with a challenge to restructure and reorganize to provide cost-effective and high-quality care to cancer patients. Oncology nurses are uniquely positioned to step into new roles emphasizing patient and family education and support. A discussion of the Health Belief Model provides an increased understanding of patient motivation and helps healthcare providers increase compliance among patients using oral therapies. This article provides an overview of the current status of oral cancer therapy in the United States and takes into consideration a historical perspective; illustrates pharmacology, indications, administration, and side effect profile through an exemplar agent; discusses potential advantages of and challenges to integration of oral therapies; and discusses alternative methods of patient and family education to improve compliance and outcome. PMID- 17413778 TI - Response shift: a theoretical exploration of quality of life following hematopoietic cell transplantation. AB - Quality of life (QOL) has become an important outcome measure for evaluating the impact of cancer therapy, especially aggressive cancer therapies such as hematopoietic cell transplantation (HCT). Despite the intense interest in examining the phenomenon of QOL, fundamental concerns remain. Most published QOL studies of HCT recipients do not state the theoretical model on which the investigation was designed. The absence of a theoretical foundation results in difficulties for healthcare professions to interpret the study's outcomes, generalize the findings and design and test theory-based interventions. Most HCT recipients report good to excellent QOL despite ongoing treatment-related sequela. This article explores the theoretical model of response shift as a means of understanding how HCT recipients maintain or improve their QOL after the treatment of life-threatening illness. Finally, a proposal for studying the QOL of HCT recipients based on the response shift model is offered, which includes a discussion of theory-based interventions. PMID- 17413779 TI - Reliability and validity of the colorectal cancer screening belief scale in Turkey. AB - Colorectal cancer is the third leading cause of cancer death in Turkey. The emphasis of the healthcare services in Turkey is on curative rather than preventive and rehabilitative approaches. Although the Ministry of Health provides many healthcare services for prevention and early detection, their availability and accessibility are very low. The purpose of this study was to test the reliability and validity of the Turkish language version of Champion's Health Belief Model Scales in measuring Turkish women's and men's beliefs about colorectal cancer. This study was carried out in Izmir, the third most populous city in Turkey. The Champion's Health Belief Model Scales was translated using a back-translation technique. A convenience sample of 470 individuals was recruited from January 2004 through March 2004. Descriptive statistics were computed for the demographic characteristics. Reliability was assessed by interpreting the item-total subscale score correlation, test-retest reliability, and Cronbach alpha coefficients. For testing the relationship between item performance and scale performance, corrected item-total correlations ranged from 0.41 to 0.79 for all 5 subscales. Cronbach alpha coefficients for the 5 subscales ranged between .54 and .88, and test-retest reliability coefficients ranged from 0.72 to 0.91. The study showed that the Turkish version of the Champion's Health Belief Model Scales has good structural characteristics and is a reliable and valid instrument that can be used for measuring beliefs related to colorectal cancer. PMID- 17413780 TI - Experiences of fatigue and self-management of Thai Buddhist cancer patients undergoing radiation therapy. AB - The purpose of this study was to describe (a) the subjective feelings of fatigue of Thai Buddhist cancer patients undergoing radiation therapy (RT) and identify possible gender differences, (b) their beliefs about the main cause of fatigue, and (c) their ways of self-management for relief of fatigue. One hundred thirty three voluntary and randomly selected patients, 55 men and 78 women, completed a questionnaire with a Thai version of the Revised Piper Fatigue Scale. After 1 week of treatment, the largest group (42.1%) of the patients had felt fatigue during the last few hours (from 1 to 5 hours). The patients perceived a mild level of fatigue, but the sensory/cognitive fatigue scores were high. Five categories of beliefs about the main cause of fatigue emerged: (a) receiving RT; (b) feelings of anxiety, depression, and fright/fear; (c) cancer; (d) long travel to hospital and long waiting time for RT; and (e) difficulty with sleep and weak body. For relief of fatigue, 5 categories of self-management emerged: (1) getting moral support from family and friends; (2) practicing religion, reciting prayers, doing merit, and meditating; (3) practicing self-care for symptomatic problems; (4) accepting the situation and doing the best of one's life; and (5) consulting with doctor and nurse. In conclusion, healthcare providers need to be concerned about RT patients' symptoms of fatigue, beliefs about causes of fatigue, and ways of self-management for relief of fatigue. They also need understanding of cultural and religious beliefs to plan holistic care. PMID- 17413782 TI - Supporting cancer patients with palliative care needs: district nurses' role perceptions. AB - The aim of this study was to examine UK district nurses' perceptions of their role in supporting palliative care cancer patients. Patients with cancer are living longer with the disease. District nurses are the largest UK workforce caring for people with cancer at home, the preferred place of care. Meeting patients' supportive and palliative care needs is complex. Little is known about district nurses' supportive role in the early phase of palliative care. Semistructured interviews were conducted with 34 district nurses. Data were analyzed thematically, with assistance from Atlas/ti. A dominant theme emerging from the interviews was ambiguity in the district nurses' supportive role in early palliative care. District nurses discussed the importance of making contact early on to support cancer patients and their families but had difficulty articulating this "support." Ambiguity, lack of confidence, and perceived skill deficits presented district nurses with dilemmas that were difficult to resolve. District nurses have great potential for meeting cancer patients' supportive and palliative care needs, a potential not currently realized. Education alone is unlikely to improve practice without an understanding of the tensions faced by district nurses in their work. Recognizing and addressing dilemmas in the everyday work of district nurses is central to moving practice forward. PMID- 17413783 TI - A pilot study of a relaxation technique for management of nausea and vomiting in patients receiving cancer chemotherapy. AB - This study was designed to determine the effect of a progressive muscle relaxation intervention on nausea and vomiting associated with anticancer chemotherapy. Subjects were 30 hematology patients who were hospitalized and received chemotherapy treatment at a large hospital in the interior of Sao Paulo, Brazil. The results indicated that progressive muscle relaxation lead to statistically significant changes in physiological and muscle conditions and in nausea and vomiting levels. Therefore, this relaxation technique may be an effective nursing intervention method to allay or alleviate nausea and vomiting in patients receiving chemotherapy. For future studies, we suggest using a control group, a homogeneous sample in terms of antiemetic and chemotherapy type and dosage, and the longitudinal following of subjects during chemotherapy. PMID- 17413785 TI - The "PrOMIS" of things to come. PMID- 17413786 TI - Natriuretic peptides: biomarkers not predictive in the intensive care unit. PMID- 17413787 TI - Back to basics, or the evolution of traumatic brain injury management since Scipione Riva-Rocci. PMID- 17413788 TI - Antimicrobial coating of central venous catheters: show me the data. PMID- 17413789 TI - Winning the cold war: inroads into implementation of mild hypothermia after cardiac arrest in adults from the European Resuscitation Council Hypothermia After Cardiac Arrest Registry Study Group. PMID- 17413790 TI - In search of the elusive sleep in critically ill and mechanically ventilated patients. PMID- 17413791 TI - Welcoming a new era of hemodynamic monitoring: expanding from the macro to the microcirculation. PMID- 17413792 TI - High-mobility group box 1 (HMGB1) as a potential therapeutic target in sepsis- more questions than answers. PMID- 17413793 TI - Improving patient safety reporting systems. PMID- 17413794 TI - Should we talk to patients (and their families) about God? PMID- 17413795 TI - Measuring up. PMID- 17413796 TI - Adoption of sepsis bundles in the emergency room and intensive care unit: A model for quality improvement. PMID- 17413797 TI - Interleukin-1 in thermal injury: what is next? PMID- 17413798 TI - Carbon monoxide: venom, endogenous mediator, or therapeutic agent? PMID- 17413799 TI - Are we finally taming inflammation? PMID- 17413800 TI - Sleeping Beauty: a tale still with an uncertain happy ending after head trauma. PMID- 17413801 TI - Evaluation of short-term consequences of hypoglycemia in an intensive care unit. PMID- 17413802 TI - Tight glucose control and hypoglycemia: should we bother? PMID- 17413803 TI - Are the fat really fit? PMID- 17413805 TI - Effects of saline or albumin resuscitation on acid-base status and serum electrolytes. PMID- 17413809 TI - Hypoglycemia risk: a cause for concern in the intensive care unit hyperglycemia control debate. PMID- 17413811 TI - From the editor ... health disparities research. PMID- 17413812 TI - Social and environmental factors: interviews of women with incarcerated partners. AB - A qualitative study was conducted to determine factors influencing how women with incarcerated partners participate in health research. Interviews were conducted with 10 women with male incarcerated partners and analyzed using content analysis. Factors that influenced whether women participated in research included their knowledge of health-related research and the conditions under which the interviews were conducted (eg, the presence of children, how information obtained is used). The notion of "location" is described as it pertains to how women maintain relationships with their incarcerated partners. Future research to examine how incarceration affects partner health and family relationships is required. PMID- 17413813 TI - Influences on the mental health of children placed in foster care. AB - Qualitative research methods were used to explore stakeholder perceptions of various influences on foster children's health. Semistructured interviews were conducted with focus groups of foster children, foster parents, and foster care professionals. Interview data were analyzed using content analysis procedures. Themes emerging from this analysis focused primarily on the foster children's mental health, including difficulties associated with perceiving oneself as being different, the children's need for coping strategies, problems encountered with the foster care system, transitions between foster homes, and the need for medical homes. Stakeholder groups recognized the necessity of mutual support for one another and proposed potential solutions for addressing concerns. PMID- 17413815 TI - Korean American maternal acceptance-rejection, acculturation, and children's social competence. AB - The goal of this study was to examine the relationship between maternal acceptance-rejection and children's social competence and the role that maternal acculturation (ie, American orientation, Korean orientation) played in this relationship in a sample of 53 Korean American mothers. Self-report data were analyzed using Pearson correlations and multiple hierarchical regressions. Low maternal acceptance-rejection was positively related to children's low social competence. Mothers' American orientation had a moderating effect on the relationship between maternal acceptance-rejection and children's social competence. Findings indicate the importance of providing parenting guidelines to mothers who are low on both acceptance-rejection and American orientation. PMID- 17413816 TI - Breastfeeding patterns in a community of Native Hawaiian mothers participating in WIC. AB - Although Hawaii has high breastfeeding initiation rates (89%), Native Hawaiian WIC participants have much lower initiation (64%) rates. Little is known about why these disparities occur. The study's aim was to describe the breastfeeding patterns of Hawaiian/part-Hawaiian women enrolled in the WIC who had initiated breastfeeding. Retrospective descriptive data (N=200) were gathered from WIC records. Descriptive and parametric statistics with univariate and multivariate analysis of breastfeeding patterns were completed. Mothers exclusively breastfeeding at initiation weaned significantly later and were significantly more likely to breastfeed for 6 months than were mothers who partially breastfed. Practice and policy implications of these findings are discussed. PMID- 17413817 TI - Correlates of physical activity in adults with mobility limitations. AB - This study identified the correlates of objectively and subjectively measured physical activity in adult wheelchair users. Fifty participants wore an activity monitor for a week and completed a questionnaire about factors associated with physical activity. Objectively measured activity correlated significantly with body mass index. Subjectively measured activity correlated significantly with age, stage of change, health, healthcare providers discussing exercise, and social support for exercise. Research on the effect of body mass index on activity in this population is needed. Intervention planners should plan programs that place emphasis on modifying the social environment (including healthcare providers) and removing attitudinal barriers. PMID- 17413818 TI - From primetime to paradise: the lived experience of OCD in Hawaii. AB - The primary aim of this qualitative study was to describe the lived experiences of men and women in Hawaii who were diagnosed with obsessive-compulsive disorder, and in so doing to provide an avenue for exploring health disparities involving this illness. Ten in-depth interviews revealed distinguishable similarities and differences in experiences of obsessive-compulsive disorder and suggested ways of understanding these similarities and differences. This research emphasized that in order to address disparities between the diagnosis and treatment of obsessive compulsive disorder, as compared to other neurobiological disorders, we must meet the critical need for education about this illness among clients, families, and clinicians. PMID- 17413819 TI - Occupational hazards and risks faced by Chinese immigrant restaurant workers. AB - Historically, Chinese immigrants to the United States have worked in restaurants to support their families. Hazards and risks associated with this population's work in restaurants are underrepresented in the literature. This ethnographic study used interviews, participant-observations, and follow-up focus sessions with 18 immigrants from China, Hong Kong, and Taiwan to identify potential physical, biological, enviromechanical, chemical, and psychosocial hazards that they face. Psychosocial hazards were most often mentioned by the participants; biological hazards were not identified in the data. Practice and research implications for addressing health disparities in this population are discussed. PMID- 17413820 TI - Simple solutions to complex issues: minimizing disparities in childhood immunization rates by providing walk-in shot clinic access. AB - The aims of this project were to minimize organizational barriers and increase access to immunizations for children aged 6 months to 21 years in a multiethnic community health center in Honolulu. The intervention consisted of opening a "walk-in" shot clinic (WISC), run by a nurse practitioner, 2 evenings per week and Saturdays. Between January and July 2005, 351 clients accessed the clinic, with 774 immunizations administered. Clinic satisfaction measures were excellent. Up-to-date immunization status for all clients improved significantly. The WISC is an effective and customer-friendly way to improve childhood and adolescent immunization rates in a community health center setting. PMID- 17413823 TI - A protocol for the urine/dipstick pad method. PMID- 17413824 TI - Descriptive study of peristomal complications. PMID- 17413826 TI - Qualitative research proposals-part II: conceptual models and methodological options. PMID- 17413827 TI - Moisture vs pressure: making sense out of perineal wounds. PMID- 17413828 TI - An economic evaluation of four skin damage prevention regimens in nursing home residents with incontinence: economics of skin damage prevention. AB - PURPOSE: To determine the cost and efficacy of 4 different regimens of incontinence-associated dermatitis (IAD) prevention in nursing home residents. METHODS: A multi-site open-label quasi-experimental study was conducted in 16 nursing homes stratified by location in 1 of 4 regions of the United States and randomly selected. In 3 of the 4 regimens, a moisture barrier ointment or cream of different compositions was applied after each episode of incontinence, and in 1 regimen, a polymer-based barrier film was applied 3 times per week. All regimens used a pH-balanced moisturizing cleanser. Time and motion measures were documented for the amount of skin care products used; the number, type, and time of caregivers performing IAD prevention care; and the number and type of supplies used. Rates of incontinence in each nursing home were determined during a 3-day surveillance period. RESULTS: A total of 1,918 nursing home residents were screened, and 51% (n = 981) qualified for prospective surveillance of incontinence dermatitis; the majority were female (80.1%) and elderly (96% > or = 65 years old). A total of 78.6% (771/981) of the participants were incontinent of both urine and feces. Compared to the 3 regimens in which a barrier was applied after each episode of incontinence, the use of a regimen in which a barrier film was applied 3 times weekly had significantly lower costs for the barrier product, labor associated with barrier application, and total cost, which included products, labor, and supplies. There were also savings in total product (cleanser and barrier) and total labor costs. CONCLUSIONS: The use of a defined skin care regimen that includes a cleanser and a moisture barrier is associated with a low rate of IAD in nursing home residents who are incontinent. Use of a polymer skin barrier film 3 times weekly is effective for preventing incontinence-associated skin breakdown and can provide significant cost savings. PMID- 17413830 TI - Prevention and treatment of moisture-associated skin damage (maceration) in the periwound skin. PMID- 17413831 TI - Treatment of incontinence-associated dermatitis (diaper rash) in a neonatal unit. PMID- 17413833 TI - The effect of a pressure ulcer prevention program and the bowel management system in reducing pressure ulcer prevalence in an ICU setting. AB - The Braden score is a reliable predictor of a patient's potential for developing pressure ulcers. Moisture is 1 of 6 indicators considered when calculating a Braden score. Efforts to reduce the effects of moisture and enzymatic activity on skin from incontinent diarrhea have employed various types of nonstandardized and nonapproved approaches. Rigid rectal tubes and catheters designed for other purposes are among the devices being utilized. These devices may put the patient at risk for rectal necrosis and loss of sphincter tone, and their use has produced no demonstrable positive outcomes. Additionally, their use is limited to those patients who are experiencing active diarrhea. The Zassitrade mark Bowel Management System (BMS) developed by Zassi Medical Evolutions, Inc., has produced promising outcomes. While the BMS is useful in managing patients with diarrhea, it can also be used to modify formed stool in patients who have wounds that are likely to become contaminated with incontinent feces, or who are too hemodynamically unstable to turn for lengthy or frequent incontinence care. PMID- 17413835 TI - Evaluation of a protocol for prevention of facility-acquired heel pressure ulcers. AB - OBJECTIVE: We sought to determine whether identification of comorbidities (in addition to daily Braden scale assessment and early aggressive pressure reduction interventions) would impact the development of facility-acquired pressure ulcers (FAPU) of the heel. METHODS: Patients admitted to a 333-bed community hospital located 20 miles west of Chicago. Overall, 70 medical records were reviewed, while 242 adult patients (46 in the intervention groups and 196 in the control groups) and 24 nurses participated in the study. The study was conducted in 4 phases, including a retrospective chart audit, 2 prospective interventions, and a product evaluation. Phase 1 entailed a retrospective chart review of patients who were admitted with or developed heel pressure ulcers over a 2-year period to identify factors most predictive of skin breakdowns. Phase 2 included two 10-day aggressive assessment and tailored intervention periods. Phase 3 involved prevalence day assessments of effectiveness of interventions as compared to patients on control nursing units. Phase 4 involved a staff survey comparing effectiveness and satisfaction of the current and trial heel pressure-relieving products. FINDINGS: Risk factors for development of heel ulcers during hospital stay or prior to admission included type 2 diabetes mellitus (T2DM), peripheral vascular disease (PVD), low albumin, and Braden scale score. During implementation of phase 2 (two 10-day assessment and tailored intervention periods), no heel FAPU were associated with the intervention. During the phase 3 FAPU prevalence assessment, 3 patients had facility-acquired heel ulcers; 2 were in the control group and would have met inclusion criteria if included in the intervention. One patient was in the intervention group but was not following the protocol. During phase 4, there were significant preferences for the trial heel pressure relief device on effectiveness, as well as patient and staff satisfaction. CONCLUSION: A pressure ulcer prevention protocol that incorporated accurate assessment of risk factors (Braden Scale and comorbidities) with frequent documentation of heel skin integrity had a positive impact on the incidence of heel FAPU. In addition, early aggressive implementation of pressure reducing and pressure-relieving devices was effective in reducing FAPU rates. PMID- 17413836 TI - Effectiveness of a honey dressing for healing pressure ulcers. AB - OBJECTIVE: To compare the effect of a honey dressing vs an ethoxy-diaminoacridine plus nitrofurazone dressing in patients with pressure ulcers. DESIGN: This 5-week randomized clinical trial evaluated the effect of a honey dressing on pressure ulcer healing. SETTING AND SUBJECTS: Thirty-six patients with a total of 68 stage II or III pressure ulcers referred from a university hospital in Izmir were enrolled in the study. Twenty-six subjects completed the trial. INSTRUMENTS: Ulcers were measured with acetate tracings and Pressure Ulcer Scale for Healing (PUSH) evaluations. METHODS: Fifteen patients with 25 pressure ulcers were treated with honey dressings, and 11 patients with 25 pressure ulcers were treated with ethoxy-diaminoacridine plus nitrofurazone dressings. Wound healing was assessed weekly using the PUSH tool, version 3.0. The primary outcome measure was the change in PUSH tool scores in each group at 5 weeks. RESULTS: The two groups were statistically similar with regard to baseline and wound characteristics. After 5 weeks of treatment, patients who were treated by honey dressing had significantly better PUSH tool scores than subjects treated with the ethoxy-diaminoacridine plus nitrofurazone dressing (6.55 +/- 2.14 vs 12.62 +/- 2.15, P < .001). CONCLUSION: By week 5, PUSH tool scores showed that healing among subjects using a honey dressing was approximately 4 times the rate of healing in the comparison group. The use of a honey dressing is effective and practical. PMID- 17413837 TI - Advances in negative pressure wound therapy: the VAC instill. AB - Acute and chronic wounds affect millions of patients. Negative pressure wound therapy (NPWT) has been available for several decades, and it has significantly impacted treatment of multiple types of wounds. The vacuum- assisted closure (VAC) system has recently been modified, allowing intermittent instillation of fluids into wounds. A description of the system, guidelines for use and novel application techniques developed by the author are discussed. PMID- 17413838 TI - Development and preliminary validation of the Menopause Symptoms Treatment Satisfaction Questionnaire (MS-TSQ). AB - OBJECTIVE: The Menopause Symptoms Treatment Satisfaction Questionnaire, an eight item questionnaire with a 4-week recall period, was developed to assess women's satisfaction with treatment for symptoms associated with menopause. We describe the development and initial testing of the scale. DESIGN: Following standard instrument-development procedures, focus groups were conducted with menopausal women experiencing hot flushes to generate potential constructs. Multiple items were drafted to address each construct. An iterative process of cognitive testing, item revision, and item reduction was followed to identify the most appropriate items and optimal response scales. The psychometric validation of the questionnaire used data collected through a multicenter, randomized, double blind, placebo-controlled study including 543 postmenopausal women. Psychometric analyses were conducted to explore potential item reduction and to address questionnaire scaling and scoring. Internal consistency reliability, construct validity, and discriminant validity of the new scale were also examined. RESULTS: The questionnaire includes items addressing the control of daytime and nighttime hot flushes; effects of treatment on sleep, mood, libido, and cognition; medication tolerability; and overall satisfaction. Correlation analyses indicated that the items are related to each other without being overly redundant and that the item set is best described using a one-factor model. The subsequent scale score demonstrated sound internal consistency reliability, strong construct validity, and good discriminant validity. CONCLUSIONS: The results of the development and initial validation are favorable. It is expected that the questionnaire will prove to be a worthwhile tool for assessing women's satisfaction with treatment for menopausal symptoms. PMID- 17413839 TI - Soy isoflavone supplementation and fasting serum glucose and lipid profile among postmenopausal Chinese women: a double-blind, randomized, placebo-controlled trial. AB - OBJECTIVE: Long-term effects of soy-derived isoflavones on lipids remain uncertain, and few data are available on their effects on glycemic control. We examined the effects of isolated soy germ isoflavones on the changes in fasting glucose (FG) and lipids. DESIGN: A double-blind, randomized, placebo-controlled trial was conducted in 203 postmenopausal Chinese women aged 48 to 62 years. They were randomly assigned to receive daily doses of 500 mg calcium, and 0 mg isoflavones (placebo, n=67), 40 mg isoflavones (n=68), and 80 mg isoflavones (n=68). Serum FG, triglycerides, high-density lipoprotein, low-density lipoprotein, total cholesterol, and lipoprotein cholesterol were measured at baseline and 1 year after treatment. The primary data analysis was performed on the 203 randomized women according to the intent-to-treat principle. The last value carried forward was used for any missing data at follow-up. RESULTS: We observed moderate but significant favorable effects of soy isoflavones on the changes (P=0.012) and percentage of changes (P=0.031) in FG (analysis of variance). The 1-year mean (SD) differences of FG changes were -5.2 (-9.4 to 1.0) mg/dL (P=0.010) and -3.3 (-7.5 to 0.9) mg/dL (P=0.18) in the 40- and 80-mg isoflavone groups compared with the placebo group. We also noted a significant interaction between the treatment and baseline FG on the changes in FG (P=0.004). The isoflavone effects were much more significant in women with baseline FG 100 mg/dL or more than in those with FG less than 90 mg/dL. We observed little effect of soy isoflavones on changes in serum lipids among the treatment groups. CONCLUSIONS: One-year of soy isoflavone supplementation might have a favorable effect on FG in women, but has no significant effect on serum lipids. PMID- 17413840 TI - Breast density as a clinical entity: is it a marker for breast cancer? PMID- 17413841 TI - Endocrine disruptors and epigenetic transgenerational disease etiology. AB - Exposure to an environmental factor (e.g. endocrine disruptor) during embryonic gonadal sex determination appears to be epigenetically reprogram the male germ line and subsequently promote transgenerational adult-onset disease. Disease phenotypes resulting from this epigenetic phenomenon include testis abnormalities, prostate disease, kidney disease, tumor development, and immune abnormalities. The epigenetic mechanism is hypothesized to involve the induction of new imprinted-like DNA sequences in the germ-line to transgenerationally transmit disease phenotypes. This epigenetic transgenerational disease mechanism provides a unique perspective from which to view adult onset disease and ultimately offers new insights into novel diagnostic and therapeutic strategies. PMID- 17413842 TI - Growth regulation, imprinted genes, and chromosome 11p15.5. AB - Genomic imprinting refers to parent-of-origin-specific gene expression. Human chromosome band 11p15.5 houses a large cluster of genes that are imprinted. Dysregulation of this gene cluster is associated with the overgrowth and tumor predisposition syndrome, Beckwith-Wiedemann syndrome. Several genes in this imprinted cluster encode proteins involved in growth regulation, e.g. the paternally expressed IGF2 and the maternally expressed cell-cycle regulator cyclin dependent kinase inhibitor, CDKN1C. Disruption of imprinted gene expression can result from genetic or epigenetic alterations. Genetic alterations such as duplication, deletion, translocation, inversion, and mutation in imprinted regions have been shown to cause disease. In addition, epimutations that are extrinsic to the primary DNA sequence have also been shown to cause disease. These epimutations usually involve gain or loss of methylation at regulatory differentially methylated regions. Recently, several human diseases in addition to Beckwith-Wiedemann syndrome have been reported to have molecular alterations at chromosome 11p15.5. These include isolated hemihyperplasia, Russell-Silver syndrome, and transient neonatal diabetes mellitus. These molecular alterations and their phenotypic effects on growth are discussed. PMID- 17413843 TI - Could epigenetics play a role in the developmental origins of health and disease? AB - Following Barker's observations of an association between birth size and later adult diseases, considerable efforts have been made to define the characteristics of low birth weight groups in childhood. In this review, the phenotypic and biochemical characteristics during childhood of three low birth weight groups are summarized: children born following inviter fertilization (IVF), small for gestational age (SGA), or very premature. Each of these groups is likely to have been exposed to an adverse environment at different developmental stages. The triggers and mechanisms leading to programmed changes in growth, development, and metabolism of these groups of children have yet to be identified. Epigenetics has been proposed as a potential mechanism for these programmed changes through environmentally induced changes in gene expression. Data from animal models in which environmental, particularly nutritional, manipulation leads to changes in DNA methylation are presented. The relevance of these animal studies to IVF, SGA, and very premature children are discussed as are potential candidate genes that may have undergone epigenetic modification to alter growth and metabolism. PMID- 17413844 TI - Epigenetic regulation of neural gene expression and neuronal function. AB - The development and function of the CNS requires accurate gene transcription control in response to proper environmental signals. Epigenetic mechanisms, including DNA methylation, histone modifications, and other chromatin-remodeling events, are critically important in mediating precise neural gene regulation. This review focuses on discussing the role of DNA methylation and histone modifications in neural lineage differentiation, synaptic plasticity and neural behavior. We postulate that DNA methylation- and histone modification-mediated gene regulation is not only important for neural cell differentiation but also crucial for high-order cognitive functions such as learning and memory. PMID- 17413845 TI - Developmental origins of beta-cell failure in type 2 diabetes: the role of epigenetic mechanisms. AB - Intrauterine growth retardation (IUGR) has been linked to later development of type 2 diabetes in adulthood. An abnormal metabolic intrauterine milieu affects the development of the fetus by permanently modifying gene expression of susceptible cells. Altered gene expression persists after birth, suggesting that an epigenetic mechanism may be responsible for changes in transcription. Uteroplacental insufficiency (IUGR) is associated with hypomethylation and hyperacetylation of genomic DNA in brain and liver of IUGR fetal and juvenile rats. These findings are associated with zinc deficiency that often accompanies fetal growth retardation. Studies in the IUGR rat also demonstrate that an abnormal intrauterine environment induces epigenetic modifications of key genes regulating beta-cell development and experiments directly link chromatin remodeling to suppression of transcription. Dietary protein restriction of pregnant rats causes fetal growth retardation and is associated with hypomethylation of the glucocorticoid receptor (GR) and PPARgamma genes in liver of the offspring. It is postulated that these epigenetic changes result in the observed increase in gene expression of GR and PPARgamma. Future research will be directed at elucidating the mechanisms underlying epigenetic modifications in offspring. PMID- 17413846 TI - Epigenetics and microRNAs. AB - MicroRNAs (miRNAs) regulate protein-coding genes post transcriptionally in higher eukaryotes. Argonaute proteins are important in miRNA regulation and are also implicated in epigenetic mechanisms such as histone modifications and DNA methylation. Here, we review miRNA regulation and outline its connections to epigenetics. PMID- 17413847 TI - Metastable epialleles, imprinting, and the fetal origins of adult diseases. AB - Epigenetics is the study of the heritable changes in gene expression that occur without a change in the DNA sequence itself. These heritable epigenetic changes include chromatin folding and attachment to the nuclear matrix, packaging of DNA around nucleosomes, histone modifications, and DNA methylation. The epigenome is particularly susceptible to dysregulation during gestation, neonatal development, puberty, and old age. Nevertheless, it is most vulnerable to environmental factors during embryogenesis because the DNA synthetic rate is high, and the elaborate DNA methylation patterning and chromatin structure required for normal tissue development is established during early development. Metastable epialleles are alleles that are variably expressed in genetically identical individuals due to epigenetic modifications established during early development and are thought to be particularly vulnerable to environmental influences. The viable yellow agouti (A(vy)) allele, whose expression is correlated to DNA methylation, is a murine metastable epiallele, which has been used as an epigenetic biosensor for environmental factors affecting the fetal epigenome. In this review, we introduce epigenetic gene regulation, describe important epigenetic phenomenon in mammals, summarize literature linking the early environment to developmental plasticity of the fetal epigenome, and promote the necessity to identify epigenetically labile genes in the mouse and human genomes. PMID- 17413848 TI - The epigenetic basis of twin discordance in age-related diseases. AB - Monozygotic twins share the same genotype because they are derived from the same zygote. However, monozygotic twin siblings frequently present many phenotypic differences, such as their susceptibility to disease and a wide range of anthropomorphic features. Recent studies suggest that phenotypic discordance between monozygotic twins is at least to some extent due to epigenetic factors that change over the lifetime of a multicellular organism. It has been proposed that epigenetic drift during development can be stochastic or determined by environmental factors. In reality, a combination of the two causes prevails in most cases. Acute environmental factors are directly associated with epigenetic dependent disease phenotypes, as demonstrated by the increased CpG-island promoter hypermethylation of tumor suppressor genes in the normal oral mucosa of smokers. Since monozygotic twins are genetically identical they are considered ideal experimental models for studying the role of environmental factors as determinants of complex diseases and phenotypes. PMID- 17413849 TI - Epigenetics in reproductive medicine. AB - Imprinted genes comprise a small subset of the genome whose epigenetic reprogramming in the germ line is necessary for subsequent normal embryonic development. This reprogramming and resetting of the imprints, through an erasure/acquisition/maintenance cycle, is a subtle and tightly orchestrated phenomenon, involving specific genomic regions and methylation enzymes. Dysregulation of imprinted genes has indeed been shown to lead to several human disorders as well as to affect placental and fetal growth. There have been numerous and conflicting studies assessing the possible association of imprinting disorders with assisted reproductive techniques. This work analyzes all relevant and available reports with regard to the association between assisted reproductive techniques and imprinting disorders. It also discusses whether this possibly increased risk of imprinting disorders may be linked to specific steps of these reproductive techniques or already present in the gametes of infertile patients. A better understanding of epigenetic reprogramming in the germ line is absolutely necessary both to assess the safety of these methods and of the use of impaired spermatogenesis gametes for assisted reproduction. PMID- 17413850 TI - Epigenetics of long-range chromatin interactions. AB - DNA segments that are separated from the promoter region of a gene by many thousands of bases may nonetheless regulate the transcriptional activity of that gene. This finding has led to the investigation of mechanisms underlying long range chromatin interactions. In intermitotic cells, chromosomes decondense, filling the nucleus with distinct chromosome territories that interdigitate and intercalate with neighboring and even more distant chromosome territories. Both intrachromosomal and interchromosomal long-range associations have been demonstrated, and DNA binding proteins have been implicated in the maintenance of these interactions. A single gene may have interactions with many distant DNA segments. Genes that are monoallelically expressed, such as imprinted genes and odorant receptors, are frequently found to be regulated by these long-range interactions. These findings emphasize the importance of studying the geography and architecture of the nucleus as an important factor in the regulation of gene transcription. PMID- 17413851 TI - Epigenetic mechanisms and the mismatch concept of the developmental origins of health and disease. AB - There is now considerable evidence that elements of the heritable or familial component of disease susceptibility are transmitted by nongenomic means, and that environmental influences acting during early development shape disease risk in later life. The underlying mechanisms are thought to involve epigenetic modifications in nonimprinted genes induced by aspects of the developmental environment, which modify gene expression without altering DNA sequences. These changes result in life-long alterations in gene expression. Such nongenomic tuning of phenotype through developmental plasticity has adaptive value because it attempts to match an individual's responses to the environment predicted to be experienced. When the responses are mismatched, disease risk increases. An example of such mismatch is that arising either from inaccurate nutritional cues from the mother or placenta before birth, or from rapid environmental change through improved socioeconomic conditions, which contribute substantially to the increasing prevalence of type-2 diabetes, obesity, and cardiovascular disease. Recent evidence suggests that the effects can be transmitted to more than the immediately succeeding generation, through female and perhaps male lines. Future research into epigenetic processes may permit us to develop intervention strategies. PMID- 17413852 TI - Epigenetics and microRNAs. AB - Epigenetics is defined as mitotically and meiotically heritable changes in gene expression that do not involve a change in the DNA sequence. Two major areas of epigenetics-DNA methylation and histone modifications-are known to have profound effects on controlling gene expression. DNA methylation is involved in normal cellular control of expression, and aberrant hypermethylation can lead to silencing of tumor-suppressor genes in carcinogenesis. Histone modifications control the accessibility of the chromatin and transcriptional activities inside a cell. MicroRNAs (miRNAs) are small RNA molecules, approximately 22 nucleotides long that can negatively control their target gene expression posttranscriptionally. There are currently more than 460 human miRNAs known, and the total number is predicted to be much larger. Recently, the expression of miRNAs has been definitively linked to cancer development, and miRNA profiles can be used to classify human cancers. miRNAs are encoded in our genome and are generally transcribed by RNA polymerase II. Despite the growing evidence for their importance in normal physiology, little is known about the regulation of miRNA expression. In this review, we will examine the relationship between miRNAs and epigenetics. We examine the effects of miRNAs on epigenetic machinery, and the control of miRNA expression by epigenetic mechanisms. Epigenetics is defined as heritable changes in gene expression that do not involve a change in DNA sequence. PMID- 17413853 TI - What is the right level of DHA in the infant diet? Commentary on article by Hsieh et al. on page 537. PMID- 17413854 TI - Ashley's case. PMID- 17413855 TI - Assessment of infant brain development with frequency-domain near-infrared spectroscopy. AB - This is the first report to demonstrate quantitative monitoring of infant brain development with frequency-domain near-infrared spectroscopy (FD-NIRS). Regionally specific increases in blood volume and oxygen consumption were measured in healthy infants during their first year. The results agree with prior PET and SPECT reports; but, unlike these methods, FD-NIRS is portable and uses nonionizing radiation. Further, new information includes the relatively constant tissue oxygenation with age and location, suggesting a tight control between local oxygen delivery and consumption in healthy infants during brain development. FD-NIRS could become the preferred clinical tool for quantitatively assessing infant brain development. PMID- 17413856 TI - Endothelin-1 mediates hypoxia-induced increases in vascular collagen in the newborn mouse lung. AB - Endothelin-1 (ET-1) mediates hypoxia-mediated pulmonary vascular remodeling (HPVR), and endothelin-A receptor (ET-AR) blockade prevents HPVR in newborn mice. Our objective was to determine postnatal effects of chronic hypoxia and/or ET-AR blockade on lung ET-1, ET-AR, ET-BR, and vascular collagen and elastin. Newborn C57BL/6 mice (n = 6-8/gp) given either BQ610 (ET-AR blocker) or vehicle were exposed to air or hypoxia (12% O2) from birth for 1, 3, or 14 d. Lung ET-1 was assessed by ELISA, and ET-AR and ET-BR by immunohistochemistry. Vascular collagen and elastin were assessed by quantitative image analysis. ET-1, ET-AR, ET-BR, collagen I and III, and tropoelastin mRNA levels were assessed by real-time quantitative RT-PCR. We observed that: 1) hypoxia attenuated the normal postnatal decrease in ET-1 and collagen content; 2) ET-AR blockade reduced collagen independent of O2; 3) hypoxia increased elastin mRNA expression and attenuated the normal postnatal decrease in elastin content; and 4) BQ610 reduced elastin mRNA but not elastin content. We conclude that, in neonatal mice, hypoxia attenuates normal postnatal decreases in ET-1, vascular collagen, and elastin. ET AR blockade reduces collagen fiber area but not mRNA, and does not decrease elastin despite reducing its expression. PMID- 17413857 TI - The influence of moderate and high dietary long chain polyunsaturated fatty acids (LCPUFA) on baboon neonate tissue fatty acids. AB - Docosahexaenoic acid (DHA) and arachidonic acid (ARA) are now common ingredients in commercial infant formulas, however, the optimal levels have not been established. Our previous data showed that the current amount of DHA in U.S. term formulas, 0.3%w/w, is insufficient to normalize cerebral cortex DHA to levels in breastfed baboon neonate controls (Diau et al.: BMC Medicine 3: 11, 2005). Here, we report on the influence of higher formula DHA levels on 12-wk-old full-term baboon CNS and visceral organs. Fourteen nursery-reared baboons were randomized to one of three diets: control (C, no DHA-ARA); moderate LCPUFA (L, 0.33%DHA 0.67%ARA); high LCPUFA (L3, 1.00%DHA-0.67%ARA). DHA increased significantly in liver, heart, and plasma (all C < L < L3), RBC (C < L, L3), and CNS regions: precentral gyrus (C < L < L3), frontal cortex, inferior and superior colliculi, globus pallidus, and caudate (all C < L, L3). These data extend previous observations indicating that 1) tissue DHA is more sensitive to diet than ARA; 2) cerebral cortex DHA increases with higher levels of DHA than in present commercial formulas; and 3) basal ganglia and limbic system DHA saturate with levels of DHA currently available in formulas. These results imply that higher levels of DHA are necessary to normalize cortex DHA to those found in breastfed animals. PMID- 17413858 TI - Antimicrobial components of the neonatal gut affected upon colonization. AB - Antimicrobial peptides (AMP) produced throughout our body are important effectors in the defense barrier of innate immunity. Here, we have analyzed antimicrobial activity and polypeptide composition of meconium versus neonatal feces to address the development of antimicrobial defense of the neonatal gut. Extracts of meconium exhibited antimicrobial activity against Bacillus megaterium, Escherichia coli, and group B streptococci (GBS) but not against the yeast Candida albicans. Extracts of neonatal feces were found to possess low activity against E. coli, GBS, and C. albicans. However, the anti-B. megaterium activity was higher in the fecal extracts than in meconium. All activities were reduced or abolished when salt was added to the antimicrobial assay. The AMP cathelicidin LL 37, alpha-defensin HNP-1-2, alpha-defensin HD 5, and lysozyme were identified in both meconium and fecal extracts. In addition, HNP-3 and a fragment of azurocidin were found in meconium, whereas the holoprotein azurocidin was detected in feces. In meconium, histones H2A and H4 were isolated and identified by their antimicrobial activity. Notably, LL-37 and lysozyme were found at significantly higher levels in feces than in meconium. Our findings reveal that meconium and feces contain AMP, acting in the defense of the neonatal gut, and may be implicated in the control of the initial colonization. PMID- 17413859 TI - Cytomegalovirus infection of human syncytiotrophoblast cells strongly interferes with expression of genes involved in placental differentiation and tissue integrity. AB - The principle route of acquisition of cytomegalovirus (CMV) for the fetus is believed to be via the placenta. We subjected purified cytotrophoblast cells obtained from full-term placentas to CMV infection and examined placental gene expression using microarray analyses. Cytotrophoblast cells purified from term placentas differentiated in vitro into a multinucleated syncytium that could be productively infected with CMV, with peak virus titers of approximately 10 plaque forming units (PFU)/mL identified in supernatants at late time points postinoculation. Infected syncytiotrophoblast cells expressed CMV-specific transcripts and proteins, as demonstrated by Northern blot and immunofluorescence assays. Microarray analyses revealed that CMV infection strongly and reproducibly altered trophoblast gene expression, elevating expression of mitotic cell cycle genes, and repressing expression of genes associated with trophoblast differentiation, particularly those associated with formation and stabilization of the extracellular matrix. We conclude that purified, differentiated syncytiotrophoblasts are permissive for CMV replication. Infection of these cells induces significant perturbations in trophoblast transcription. An improved understanding of the molecular events that occur during CMV infection of trophoblasts could provide insights into interventions that might prevent or minimize congenital transmission. PMID- 17413860 TI - Maternal stress during pregnancy predisposes for iron deficiency in infant monkeys impacting innate immunity. AB - The influence of maternal stress during pregnancy on the postpartum iron status and immune maturation of infants was investigated in a nonhuman primate model. Forty infant rhesus monkeys were generated from two types of disturbed pregnancies, early or late gestation stress, and compared with 24 undisturbed controls. Prenatal stress increased the prevalence and magnitude of iron deficiency (ID) as the infants' growth-related demands for iron exceeded dietary intake from breast milk. At 4-6 mo of age, the emergence of ID significantly accentuated an effect of prenatal stress on natural killer cell activity, an important component of innate immunity. These findings indicate that maternal stress, especially early in pregnancy, should be added to the list of risk factors that warrant closer scrutiny of hematological profiles in fast-growing babies. PMID- 17413861 TI - Influence of labor on neonatal neutrophil apoptosis, and inflammatory activity. AB - Neutrophil apoptosis is impaired in neonates, and this contributes to prolonged inflammation and tissue injury in infants after infection or trauma. In the present studies, we investigated whether labor generates mediators that further suppress apoptosis. We found that neutrophil apoptosis was reduced in neonates exposed to labor, when compared with infants delivered by cesarean section before labor. This was not due to alterations in caspase-3 or inhibitor of apoptosis protein-2 (IAP-2). In contrast, labor primed neutrophils to express tumor necrosis factor alpha (TNF-alpha), suggesting that proinflammatory mediators contribute to reduced apoptosis after labor. Eicosanoids generated via cyclooxygenase-2 (Cox-2) and lipoxygenase (Lox) also regulate neutrophil apoptosis. 15-Lox, which generates proapoptotic lipoxins, but not Cox-2, was greater in neutrophils before labor, relative to cells exposed to labor. Anti inflammatory eicosanoids exert their effects in part via peroxisome proliferator activated receptor gamma (PPAR-gamma). Expression of gelatinase-associated lipocalin and catalase, two markers of PPAR-gamma activity, were increased in neonatal neutrophils before labor, relative to cells exposed to labor. These findings suggest that the anti-inflammatory environment is maintained before labor, in part, by eicosanoids. Although increased neutrophil longevity after labor is important for host defense in the immediate newborn period, it may contribute to inflammatory or oxidative injury in susceptible infants. PMID- 17413862 TI - Inhaled nitric oxide: effects on cerebral growth and injury in a baboon model of premature delivery. AB - Inhaled nitric oxide (iNO) enhances ventilation in very preterm infants, but the effects on the brain remain uncertain. We evaluated the impact of iNO on brain growth and cerebral injury in a premature baboon model. Baboons were delivered at 125 d of gestation (term 185 d of gestation) and ventilated for 14 d with either positive pressure ventilation (PPV) (n = 7) or PPV + iNO (n = 8). Brains were assessed histologically for parameters of development and injury. Compared with gestational controls (n = 7), brain and body weights were reduced but brain-to body weight ratios were increased in all prematurely delivered (PD) animals; the surface folding index (SFI), was reduced in PPV but not PPV + iNO animals. Compared with controls, the brain damage index was increased (p < 0.05) in both cohorts of PD animals. There was no difference between ventilatory regimens, however, in 25% of animals with iNO therapy, there were organized hematomas in the subarachnoid space. Overall, iNO did not alter the extent of brain damage but did result in the presence of hematomas. These results do not confirm any protective or major injurious effect of nitric oxide therapy on the developing brain. PMID- 17413863 TI - Detection of ureaplasma DNA in endotracheal samples is associated with bronchopulmonary dysplasia after adjustment for multiple risk factors. AB - Microorganisms are hypothesized to contribute to the pathogenesis of bronchopulmonary dysplasia (BPD) in very low birth weight (VLBW) infants. This hypothesis remains controversial. We sought to determine whether endotracheal colonization with Ureaplasma sp., adenovirus, or Chlamydia sp. increases the risk of BPD. Intubated VLBW infants were included. Polymerase chain reaction (PCR) analysis was used to detect Ureaplasma sp., adenovirus, and Chlamydia sp. The outcome measure was BPD or death due to lung disease. Detection of microorganisms was compared between subjects with and without BPD. Logistic regression was used to control for covariates. Of 139 subjects, 33 (25%) screened positive for Ureaplasma sp., 22 of 136 (16%) were positive for adenovirus; eight of 133 (6%) were positive for Chlamydia sp. At 36 wk postmenstrual age, 14 patients had died, 68 (57%) had BPD. Detection of Ureaplasma sp. was associated with BPD or death (p < 0.001); adenovirus (p = 0.52) and Chlamydia sp. (p = 0.33) were not. Controlling confounding factors, the odds ratio for Ureaplasma sp. and BPD or death was 4.2 (95% CI 1.03, 17). In our population, detection of Ureaplasma sp., but not adenovirus or Chlamydia sp. was associated with BPD or death due to lung disease. PMID- 17413864 TI - Relationship between opioid therapy, tissue-damaging procedures, and brain metabolites as measured by proton MRS in asphyxiated term neonates. AB - To examine the effects of opioid and tissue-damaging procedures (TDPs) [i.e. procedures performed in the neonatal intensive care unit (NICU) known to result in pain, stress, and tissue damage] on brain metabolites, we reviewed the medical records of 28 asphyxiated term neonates (eight opioid-treated, 20 non-opioid treated) who had undergone magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy (MRS) within the first month of life as well as eight newborns with no clinical findings of asphyxial injury. We found that lower creatine (Cr), myoinositol (Ins), and N-acetylaspartate (NAA)/choline (Cho) (p < or = 0.03) and higher Cho/Cr and glutamate/glutamine (Glx) Cr (p < or = 0.02) correlated with increased TDP incidence in the first 2 d of life (DOL). We also found that occipital gray matter (OGM) NAA/Cr was decreased (p = 0.03) and lactate (Lac) was present in a significantly higher amount (40%; p = 0.03) in non opioid-treated neonates compared with opioid-treated neonates. Compared with controls, untreated neonates showed larger changes in more metabolites in basal ganglia (BG), thalami (TH), and OGM with greater significance than treated neonates. Our data suggest that TDPs affect spectral metabolites and that opioids do not cause harm in asphyxiated term neonates exposed to repetitive TDPs in the first 2-4 DOL and may provide a degree of neuroprotection. PMID- 17413865 TI - Effects of delayed pubertal development, nutritional status, and disease severity on longitudinal patterns of growth failure in children with sickle cell disease. AB - Previous studies of children with sickle cell disease (SCD) reported poor growth and delayed maturation. However, the prevalence, magnitude, and correlates of suboptimal growth remain poorly understood. A prospective longitudinal study was undertaken to determine the effects of disease severity and nutritional status on growth, an indicator of childhood well-being. Children, birth to 18 y of age, with SCD-SS were evaluated annually for 4 y. Growth, nutritional status, skeletal and sexual maturation, disease severity, dietary intake, and maternal education were assessed. In this sample of 148 children (78 females), growth in height, weight, or body mass index declined in 84% of subjects; 38% fell below the 5th percentile in one or more measures. Puberty was delayed 1 to 2 y, and median age at menarche was 13.2 y. Skeletal age was delayed by 0.7 +/- 1.4 y overall and by 1.3 +/- 1.5 y in children 10 to 15 y old. Height status declined over time and was positively associated with advancing puberty and hematological measures in girls, and nutritional status in girls and boys. Growth failure and maturational delay remain significant chronic problems in children with SCD-SS and are related to potentially modifiable factors such as nutritional status. PMID- 17413866 TI - Meeting report on the 3rd International Congress on Developmental Origins of Health and Disease (DOHaD). AB - Developmental origins of health and disease (DOHaD) focuses on the earliest stages of human development, and provides a novel paradigm to complement other strategies for lifelong prevention of common chronic health conditions. The 3 International Congress on DOHaD, held in 2005, retained the most popular features from the first two biannual Congresses, while adding a number of innovations, including increased emphasis on implications of DOHaD for the developing world; programs for trainees and young investigators; and new perspectives, including developmental plasticity, influences of social hierarchies, effects of prematurity, and populations in transition. Emerging areas of science included, first, the controversial role of infant weight gain in predicting adult obesity, diabetes, and cardiovascular disease. Second, in the era of epidemic obesity, paying attention to the over-nourished fetus is as important as investigating the growth retarded one. Third, environmental toxins appear to have abroad range of long-lasting effects on the developing human. Fourth, epigenetic mechanisms could unite several strands of human and animal observations, and explain how genetically identical individuals raised in similar postnatal environments can nonetheless develop widely differing phenotypes. Improving the environment to which an individual is exposed during development may be as important as any other public health effort to enhance population health world wide. PMID- 17413867 TI - The IL-10 (-627 A/C) promoter polymorphism may be associated with coronary aneurysms and low serum albumin in Korean children with Kawasaki disease. AB - Kawasaki disease (KD) is an acute febrile vasculitic syndrome of unknown etiology that preferentially affects the coronary artery. Interleukin-10 (IL-10) is a key proinflammatory cytokine, and a polymorphism near the major transcriptional start site of the IL-10 gene was shown to influence IL-10 production in vitro. This study investigated the association of the IL-10 promoter polymorphism with KD and its clinical parameters in Korean children. A total of 194 children with congenital heart disease (CHD) and 95 children with KD were included in this study. IL-10 (-627 A/C) polymorphism genotypes were determined using the single base extension method. There was no difference in the allele frequencies of IL-10 (-627 A/C) polymorphism between CHD children and KD children. KD children with one or two copies of the IL-10 (-627C) allele showed significantly lower albumin levels (p = 0.020) and higher frequencies of early coronary artery aneurysm [62.22% versus 37.78%, adjusted odds ratio (aOR) = 3.50, 95% confidence interval (CI): 1.50-8.16] compared with KD children with the common IL-10 (-627A) allele. These findings suggest that the IL-10 (-627 A/C) promoter polymorphism might be a genetic marker for the risk of early coronary artery complication in KD. PMID- 17413868 TI - Effect of late-onset sepsis on energy expenditure in extremely premature infants. AB - The purpose of this study was to compare total energy expenditure (TEE) in extremely premature infants during and after an episode of sepsis. We hypothesized that TEE in the sepsis group (SEP) would be higher during the septic period and higher than an age-matched control group (CTL). We further hypothesized that the TEE of the SEP group during the recovery period would be similar to that of the CTL group. The doubly labeled water method was used to determine TEE in both groups. Infant characteristics were as follows: SEP group, n = 10, gestation = 26 +/- 1 wk, birth weight = 854 +/- 218 g; CTL group, n = 10, gestation = 26 +/- 1 wk, birth weight = 880 +/- 158 g. TEE of the SEP group during the septic period was significantly greater than during the recovery period (96 +/- 25 kcal/kg/d versus 55 +/- 17 kcal/kg/d) and significantly greater than the CTL group during the first study period (96 +/- 25 kcal/kg/d versus 67 +/- 12 kcal/kg/d). TEE in the SEP group during the recovery period was similar to the CTL group. These increases in TEE may contribute to impaired growth and need to be considered when providing nutritional support for extremely premature infants. PMID- 17413869 TI - Impact of serum adiponectin concentration on birth size and early postnatal growth. AB - In term neonates, the adiponectin concentration is higher than it is in adults. To determine the relationship between adiponectin and early neonatal growth in a cohort study. Fifty-two neonates at term were studied. Serum adiponectin concentrations, body sizes, and skinfold thicknesses were measured at birth and at 1 mo of age. At birth, cord blood adiponectin concentration correlated positively with birth weight (r = 0.484, p = 0.0003), birth length (r = 0.524, p < 0.0001), and sum of the four skinfold thickness measurements (r = 0.378, p = 0.0057). In a stepwise regression, birth length was the only determinant of cord blood adiponectin concentration. However, at 1 mo of age, serum adiponectin concentration correlated with no anthropometric parameter at all. Between birth and 1 mo of age, the individual change in adiponectin concentration correlated negatively with birth weight. Thus, serum adiponectin concentrations in cord blood have a strong relationship to birth length rather than to body fatness, and this relationship is not demonstrated in 1-mo-old infants. These results imply that hormonal, substrate, or other mechanisms that regulate the relationship between body composition and growth in fetal life are different from those governing these relationships in early postnatal life. PMID- 17413870 TI - Effect of topical emollient treatment of preterm neonates in Bangladesh on invasion of pathogens into the bloodstream. AB - Topical emollient therapy may reduce the incidence of serious infections and mortality of preterm infants in developing countries. We tested whether emollient therapy reduced the burden of pathogens on skin and/or prevented bacterial translocation. Neonates <33 wk gestational age were randomized to treatment with sunflower seed oil (SSO) or Aquaphor or the untreated control group. Skin condition score and skin cultures were obtained at enrollment and on d 3, 7, and weekly thereafter, and blood cultures were obtained for episodes of suspected nosocomial sepsis. For analysis, blood cultures were paired with skin cultures obtained 0-3 d before the blood culture. Skin condition scores at 3 d were better in patients treated with either emollient compared with untreated controls; however, skin flora was similar across the groups. The SSO group showed a 72% elevated odds of having a false-positive (FP) skin culture associated with a negative blood culture (i.e. skin flora blocked from entry into blood) compared with the control group. Topical therapy with SSO reduced the passage of pathogens from the skin surface into the bloodstream of preterm infants. PMID- 17413871 TI - T helper 1 background protects against airway hyperresponsiveness and inflammation in guinea pigs with persistent respiratory syncytial virus infection. AB - A family history of allergy has been implicated in children who develop post bronchiolitis wheezing and asthma. In a guinea pig model of respiratory syncytial virus (RSV) lung infection, we evaluated the role of host Th1 background (either genetic or induced) on the development of a persistent infection, nonspecific airway hyperresponsiveness (AHR) and airway inflammation. Allergy resistant/T helper 1 (Th1)-skewed strain 2 guinea pigs (STR2) and cytosine phosphate guanine oligodeoxynucleotides (CpG-ODN) (Th1 stimuli) pretreated Cam Hartley guinea pigs (CH) were inoculated with RSV and compared with virus-inoculated allergy susceptible/Th2-skewed CHs and to sham-inoculated STR2 and CH, 60 d post inoculation. We measured titers of intrapulmonary RSV, lung interferon (IFN) gamma and interleukin (IL)-5 mRNA expression, AHR and airway T cells and eosinophils. All virus-inoculated groups of animals showed evidence of persistent RSV lung infection; however, Th2-skewed guinea pigs had virus-associated AHR and significantly greater levels of airway T cells and eosinophils. In conclusion, RSV can establish persistent infection of the guinea pig lung regardless of host Th1/Th2 background; however; a host Th1 background limits the extent of virus associated AHR and airway inflammation. Heterogeneity in virus-host interactions may be relevant to understanding why some children hospitalized for RSV bronchiolitis go on to develop recurrent wheezing/asthma symptoms. PMID- 17413872 TI - Altered cardiovascular control in preterm infants with bronchopulmonary dysplasia. AB - Vestibulo-mediated cardiovascular control in hazardous situations is important. Our hypothesis is that the prerequisite for sudden infant death syndrome (SIDS) is impaired vestibulo-mediated cardiovascular control. Prematurity is a risk factor for SIDS, and postnatal intermittent hypoxia may contribute to this risk. We studied heart rate (HR) and blood pressure (BP) responses in 10 infants with bronchopulmonary dysplasia (BPD) who were born at 27 +/- 2.4 (23-30) wk of gestation. Twenty healthy term infants served as controls. Cardiovascular tests were performed under polysomnographic control during slow-wave sleep (SWS) at a corrected age of 12 +/- 3.5 (7-19) wk. Control infants showed biphasic HR and BP responses to side motion with an immediate increase followed by a modest decrease and return to baseline. Compared with the controls, half of the BPD infants had altered BP responses (p < 0.005) without an early increase, followed by a more prominent decrease in BP. BPD infants also presented with a greater variability in BP responses to head-up tilts than did the controls (p < 0.001). In conclusion, these findings suggest that some BPD infants have impaired vestibular sympathoreflex-mediated cardiovascular control. This dysfunction may become critical in life-threatening situations. PMID- 17413873 TI - A novel ND3 mitochondrial DNA mutation in three Korean children with basal ganglia lesions and complex I deficiency. AB - Mitochondrial disorders have notoriously variable clinical presentations, particularly in children. A growing number of reports describe mutations in the mitochondrial DNA (mtDNA)-encoded subunits of complex I (EC 1.6.5.3) causing early-onset encephalopathy. Here, we describe two Korean siblings with childhood onset progressive generalized dystonia and one Korean child with strokelike episodes in infancy; all three had bilateral lesions of the basal ganglia and partial deficiencies of complex I. Analysis of their mtDNA revealed a novel heteroplasmic m.10197G>A mutation (A47T) in the ND3 (NADH dehydrogenase subunit 3) gene. This study underscores the importance of screening mtDNA-encoded respiratory chain structural genes, including ND3, in pediatric patients with unexplained encephalopathies. PMID- 17413874 TI - Congenital breast deformity reconstruction using perforator flaps. AB - BACKGROUND: Congenital breast deformities such as Poland syndrome, unilateral congenital hypoplasia, tuberous breast anomaly, and amastia pose a challenging plastic surgical dilemma. The majority of patients are young, healthy individuals who seek esthetic restoration of their breast deformities. Currently, both implant and autologous reconstructive techniques are used. This study focuses on our experience with congenital breast deformity patients who underwent reconstruction using a perforator flap. METHODS: From 1994 to 2005, a retrospective chart review was performed on women who underwent breast reconstruction using perforator flaps to correct congenital breast deformities and asymmetry. Patient age, breast deformity type, perforator flap type, flap volume, recipient vessels, postoperative complications, revisions, and esthetic results were determined. RESULTS: Over an 11-year period, 12 perforator flaps were performed. All cases were for unilateral breast deformities. The patients ranged from 16 to 43 years of age. Six patients had undergone previous correctional surgeries. Eight (n = 8) flaps were used for correction of Poland syndrome and its associated chest wall deformities. Four (n = 4) flaps were used for correction of unilateral breast hypoplasia. In all cases, the internal mammary vessels were the recipient vessels of choice. No flaps were lost. No vein grafts were used. All patients were discharged on the fourth postoperative day. Complications encountered included seroma, hematoma, and nipple malposition. Revisional surgery was performed in 30% of the cases. Esthetic results varied from poor to excellent. CONCLUSIONS: Perforator flaps are an acceptable choice for patients with congenital breast deformities seeking autologous breast reconstruction. Deep inferior epigastric artery (DIEP) or superficial inferior epigastric artery (SIEA) flaps are performed when adequate abdominal tissue is available; however, many young patients have inadequate abdominal tissue, thus a GAP flap can be used. Perforator flaps are a safe, reliable surgical technique. In the properly selected patient, donor-site morbidity and functional compromise are minimized, improved self-esteem is noted, postoperative pain is decreased, and excellent long-term esthetic results can be achieved. PMID- 17413875 TI - Double pedicle deep inferior epigastric perforator/muscle-sparing TRAM flaps for unilateral breast reconstruction. AB - BACKGROUND: Utilizing both rectus abdominis muscles for unilateral breast reconstruction poses significant risks for hernia or bulge formation and decreased abdominal wall strength. We have used the fascial sparing double pedicle deep inferior epigastric artery perforator (DIEP)/DIEP or DIEP/muscle sparing TRAM (MS-TRAM) flap to overcome the disadvantages of the conventional bilateral TRAM or bilateral free TRAM flaps. METHODS: Between January 1996 and March 2005, 14 double pedicle free DIEP/DIEP or DIEP/MS-TRAM flaps were performed in 14 patients for unilateral breast reconstruction. The abdominal wall was closed without mesh in all cases. RESULTS: Evaluation of results was conducted through a retrospective chart review and questionnaire. The average hospital stay was 5.4 days. The follow-up period ranged from 9 months to 10 years. There was no flap loss. There were no hernias, although 1 obese patient had lower abdominal wall bulging. Questionnaire was returned by 11 out of 14 patients, with an average rating of 4.5 (1-5), with all but 1 who would recommend it to others. None of the 11 patients had postoperative abdominal pain or back pain, and 9/11 patients returned to daily activities and/or sports. All patients that worked preoperatively returned to work postoperatively. CONCLUSIONS: Double pedicle free flaps for unilateral breast reconstruction are a safe option when autologous breast reconstruction is desired, but the volume of tissue required to build a breast exceeds the amount that could be transferred on a single pedicle flap. The double pedicle DIEP (DIEP/DIEP) and/or MS-TRAM (DIEP-MS-TRAM) flap offers good symmetric results for unilateral breast reconstruction and can minimize abdominal wall morbidity. PMID- 17413876 TI - Changes in body image and health-related quality of life following breast reduction surgery in German macromastia patients: a new tool for measuring body image changes. AB - Health insurance funds in Germany are increasingly reluctant to fund the cost for reduction mammaplasty. However, several studies have already demonstrated the beneficial effects of breast reduction on symptom relief and health-related quality of life. More specifically, the psychologic and social consequences of breast reduction surgery were also recently evaluated. Relating to the contemporary debate on financial restraint, the present article describes a follow-up study conducted in a sample of 40 patients undergoing reduction mammaplasty. The purpose of the investigation was to assess indicators of health related quality of life following reduction mammaplasty. Furthermore, the study aimed to assess body image changes and to test a new assessment instrument, Digital-Body-Photo-Test (DBPT), in comparison with a well-validated body image measure (Color-a-Person Body Dissatisfaction Test, CAPT) (concurrent validity). As hypothesized, the findings indicate significant improvements in health-related quality-of-life measures and body-image characteristics. The substantial improvement of body-image satisfaction of all body areas suggests a generalized positive effect of reduction mammaplasty on overall body image. The strong association between the DBPT and the CAPT scores in this study indicates that DBPT is an efficient and valid new tool for measuring body-image changes relating to patients' evaluations of their average satisfaction of specific body parts or areas and their overall appearance acceptance. PMID- 17413877 TI - Persistent pain following postmastectomy breast reconstruction: long-term effects of type and timing of surgery. AB - This study prospectively examined the long-term effects of type (transverse rectus abdominis musculocutaneous [TRAM] versus implant) and timing (immediate versus delayed) of postmastectomy reconstructive surgery on patient reports of pain at multiple body sites. Women (n = 205) seeking immediate or delayed breast reconstruction, choosing either expander implant or autologous tissue transfer surgical procedures, provided ratings for the presence of bodily, breast, abdominal, and back pain and abdominal tightness prior to surgery and at 2-year follow-up. At baseline, nonwhite women were more likely to undergo delayed reconstruction (P < 0.05), and women seeking delayed reconstruction had less breast pain (P < 0.001) and more back pain (P < 0.01). Multiple regression analyses, controlling for ethnicity and baseline pain, indicated that women receiving TRAM flap surgery reported more problems with abdominal pain and tightness. There was a trend for implant subjects to report more frequent problems with breast pain. These results suggest the need for heightened awareness of potential long-term pain morbidity for women undergoing TRAM flap or implant breast reconstruction. PMID- 17413878 TI - Breast implant capsules are partially composed of bone marrow-derived cells. AB - Capsular contracture is the most common complication following breast augmentation or reconstruction with implants. We recently demonstrated that bone marrow-derived cells provide fibroblasts to murine skin during wound healing. To determine if bone marrow-derived cells were the cellular source of periprosthetic capsules, we created chimeric C57BL mice containing bone marrow cells from isogeneic enhanced green fluorescent protein (EGFP) mice and implanted with a textured silicone shell implant. We found that none of the mice developed infection or capsular contracture, but day 30 capsules were composed of 26.4 +/- 6.1% EGFP cells, and day 60 capsules had 21.8 +/- 10.3% EGFP cells. Immunohistochemistry revealed a small population of EGFP cells in the capsules that were myofibroblasts. Thus, breast implant capsules are partially composed of bone marrow-derived cells and, given the potential of these cells to become myofibroblasts, may explain the cellular source of capsular contracture when it develops. PMID- 17413879 TI - Latissimus dorsi muscle and its short perforator-based skin compound free flap. AB - Some authors have described the latissimus dorsi muscle and its short perforator based skin compound flap based on the same thoracodorsal vessels. This flap procedure involves separating the skin island from the underlying latissimus dorsi muscle and rotating the skin island over the musculocutaneous perforator emerging from the latissimus dorsi muscle. As with all surgical procedures, there are various advantages and disadvantages. However, to the best of the authors' knowledge, there are no reports on the use of the flap in a consecutive series. Between 1997 and 2005, the flap was used to reconstruct below-knee structures in 26 patients (23 males and 3 females) by the 2 senior authors at the Chonnam National University Medical School. The clinical outcomes of this procedure were evaluated. Satisfactory results were obtained in most patients. However, there were 2 marginal necroses in the excessively large skin flaps, 1 partial necrosis over the distal edge of a skin flap, and 1 total flap failure caused by infection. The marginally and partially necrotized skin flaps were treated successfully with split-thickness skin grafts. However, another flap procedure was required to cover the defect in the case of flap failure. The donor sites were closed primarily in all patients, and the skin flaps were rotated between 40 degrees and 180 degrees. This flap allows the surface of the latissimus dorsi musculocutaneous flap to be expanded without additional donor morbidity. In addition, the flap procedure is safe and easy to perform. Moreover, the flap provides sufficient flexibility, even though it has less independent flap mobility than the chimeric flap. Therefore, the flap may be a convenient and reliable alternative for the reconstruction of large and irregular-shaped wounds. PMID- 17413880 TI - Biomechanical evaluation of cranial flap fixation techniques: comparative experimental study of suture, stainless steel wire, and rivetlike titanium clamp. AB - OBJECTIVE: To study the techniques of cranial flap fixation (suture, stainless steel wire, and rivetlike titanium clamp) on biomechanics. METHODS: Twenty-four cadaver craniotomy flaps were reattached with either suture, stainless steel wire, or rivetlike titanium clamp. Cranial flap fixation was timed and measured for the offset between cranial flap with craniotomy skull for every fixation technique. Load-bearing tests were performed by applying an external force until the failure of the fixation system. RESULTS: Rivetlike titanium clamp required significantly less time to fix (94 +/- 13 seconds) than either suture (172 +/- 14 seconds) or stainless steel wire (399 +/- 45 seconds). The offset of cranial flap after rivetlike titanium clamp fixation (0.20 +/- 0.12 mm) was significantly smaller than either suture (2.20 +/- 0.24 mm) or stainless steel wire (1.32 +/- 0.34 mm). Rivetlike titanium clamp was also stronger than suture and stainless steel wire in the load-bearing test. CONCLUSION: Rivetlike titanium clamp is easy to use, requires less time consumption, and shows accuracy and strength. PMID- 17413881 TI - Function of the clavicular part of the pectoralis major muscle after transplantation of its sternocostal part. AB - BACKGROUND: Muscle-sparing transplantation of the sternocostal part of the pectoralis major muscle while preserving the clavicular part of the muscle may reduce donor-site morbidity, particularly in cases where dissection of the lymph nodes of the neck has been performed. The nerve supply and motor function of the clavicular part is alleged to be preserved when the sternocostal part is transposed through the deltopectoral groove. This study aims to objectify such preservation. METHODS: Muscle activity of the remaining clavicular and abdominal muscle parts was determined electromyographically, dynamometrically, and goniometrically on 17 sides in 16 patients after head and neck reconstruction. Subjective assessment was obtained by use of a structured questionnaire. RESULTS: The electromyogram proved preservation of innervation of 16 of 17 clavicular parts. The electromyogram of the abdominal part showed no signs of denervation on 10 sides and normal, full interference patterns at maximum voluntary effort in all patients. Selective functional testing of the clavicular part revealed decreased force in horizontal adduction in 11 out of 15 patients. Still, the isolated effect of the pectoralis major transposition on shoulder function could not be objectified statistically. Subjective evaluation yielded that shoulder disability was more likely to have been correlated with loss of trapezius muscle function. CONCLUSION: We conclude that the innervation of the clavicular part, indeed, is preserved by transposition of the sternocostal part through the deltopectoral groove. This supports the validity, effectiveness, and functional acceptability of our muscle-sparing technique of partial pectoralis major transplantation. PMID- 17413882 TI - The versatility of the distally based peroneus brevis muscle flap in reconstructive surgery of the foot and lower leg. AB - Soft tissue and bone defects of the lower leg, ankle, and heel region often require coverage by local or distant flaps. The authors successfully used the distally based peroneus brevis muscle flap for the treatment of 15 patients with osteomyelitis (n = 5), melanoma (n = 1), Achilles tendon defects (n = 6), posttraumatic bone defects (n = 2), and chronic diabetic heel ulcer (n = 1). The size of the defects ranged from 6 to 60 cm. All defects were covered successfully without major complications by the muscle flap. The distally based peroneus brevis muscle represents a very reliable flap for coverage of small and moderate defects of the medial and lateral malleolus, the Achilles tendon, and the heel area. This flap offers a convincing alternative for covering defects in the distal leg region and is often preferable to the use of free flaps because the surgery is rapidly performed and does not require microsurgical expertise. PMID- 17413883 TI - Free fibula donor-site morbidity: clinical and biomechanical analysis. AB - The aim of this study was the quantitative assessment of donor-site morbidity after fibula harvest. Ten patients were identified with a unilateral fibula harvesting and healthy contralateral side. The average follow-up time was 32.3 months. The study design included standardized questionnaire; morbidity point evaluation system; orthopedic-biomechanical evaluation, including true ankle motion, clinical-functional American Orthopedic Foot and Ankle Society (AOFAS) hindfoot score, pedobarography, measurement of maximal voluntary isometric strength, and radiologic examination using an ankle osteoarthritis grading. Average donor-side morbidity score was low, with 2.6 points (range, 0 to 13). Average AOFAS hindfoot score was good, with 87.3 points (range, 74 to 100). Pedobarography showed significant decrease of the big toe pushup force. Isometric strength measurement showed significant reduction of the peroneus longus muscle. Discrete to mild medial ankle osteoarthritis was found in 70% of the cases. The overall donor-side morbidity was low, but an eversion torque deficit could initiate or support a progressive functional ankle instability and in long-term cause or accelerate a ligamentous secondary ankle osteoarthritis. PMID- 17413884 TI - A new flap for reconstruction of pilonidal sinus: lumbar adipofascial turnover flap. AB - BACKGROUND: "Adipofascial turnover flap" is a well-known procedure, but it is generally not used for the surgical treatment of pilonidal sinus disease. The "lumbar adipofascial turnover flap" has been used in this study for the reconstruction of uncomplicated pilonidal sinus disease. METHODS: Ten cases (8 male and 2 female patients) were operated on by this technique. The reconstruction was performed with the lumbar adipofascial turnover flap. The flap sizes ranged from 4 x 7 cm to 5 x 9 cm (mean, 4.5 x 8 cm), and they were elevated with length-to-base ratio below 2:1. The follow-up period was 14 to 26 months. RESULTS: Postoperative magnetic resonance imaging, computed tomography scan, and power Doppler ultrasound examinations revealed viability of the flaps in all patients. There was no distortion of anatomic landmarks in any of the cases. The esthetic results were satisfying for all patients as well. There was no recurrence in any cases. CONCLUSIONS: The hospital stay and mean time off work were shorter compared with other methods of reconstruction and there was no recurrence. We advocate that the lumbar adipofascial turnover flap is an excellent choice for reconstruction of cases with uncomplicated pilonidal sinus disease. PMID- 17413885 TI - Effect of surgically removing subcutaneous fat by abdominoplasty on leptin concentrations and insulin sensitivity. AB - The aim of this study was to identify the effect of surgically removing subcutaneous fat by abdominoplasty on leptin concentrations and insulin sensitivity. An open clinical trial with a noninterventional parallel group was carried out in 12 obese women. After randomization, 6 volunteers were selected for abdominoplasty, and the other 6 women were considered as the noninterventional group. A metabolic profile, including leptin concentrations, and insulin tolerance test to assess insulin sensitivity were performed on all volunteers before intervention or nonintervention and 40-50 days afterward. A significant reduction in body mass index (30.7 +/- 0.9 versus 29.6 +/- 0.7 kg/m; P = 0.02) and in leptin concentrations (41.3 +/- 10.6 versus 32.0 +/- 10.2 ng/mL; P = 0.02) was observed after abdominoplasty. Insulin sensitivity did not change after intervention. In conclusion, surgically removing subcutaneous fat by abdominoplasty decreased leptin concentrations, with no change in insulin sensitivity. PMID- 17413886 TI - Vascular anatomy of plantar muscles. AB - Many reports on the plantar arteries and the deep plantar arch exist, but none of them focus on the arterial pedicles of the plantar muscles. They mainly discuss the deep plantar arch, its variations, and location. This study plans to determine the location and origin of arterial pedicles of all the plantar muscles as a preliminary study for designing new flaps. The study was carried out on 20 feet from 10 cadavers aged from 35 to 67 years. After an injection of latex via popliteal arteries, dissection of the arteries was carried out under a microscope. Abductor hallucis and flexor hallucis brevis muscles receive their main blood supply from the medial plantar artery; abductor digiti minimi and flexor digiti minimi brevis muscles receive their main blood supply from the lateral plantar artery. The flexor digitorum brevis muscle receives branches from both arteries. Adductor hallucis and plantar interosseous muscles receive branches from plantar metatarsal arteries. Quadratus plantae is directly nourished from a branch of the posterior tibial artery. No distal anastomoses between the medial and lateral plantar arteries were identified, except 1 specimen in which the medial plantar artery made anastomosis with the deep plantar arch. As a result, the arterial pedicles of all the plantar muscles were defined, and based on these findings, new flaps can be planned or existing flaps can be modified. PMID- 17413887 TI - Genital sensitivity after sex reassignment surgery in transsexual patients. AB - BACKGROUND: Tactile and erogenous sensitivity in reconstructed genitals is one of the goals in sex reassignment surgery. Since November 1993 until April 2003, a total of 105 phalloplasties with the radial forearm free flap and 127 vaginoclitoridoplasties with the inverted penoscrotal skin flap and the dorsal glans pedicled flap have been performed at Ghent University Hospital. The specific surgical tricks used to preserve genital and tactile sensitivity are presented. In phalloplasty, the dorsal hood of the clitoris is incorporated into the neoscrotum; the clitoris is transposed, buried, and fixed directly below the reconstructed phallic shaft; and the medial and lateral antebrachial nerves are coapted to the inguinal nerve and to one of the 2 dorsal nerves of the clitoris. In vaginoplasty, the clitoris is reconstructed from a part of the glans penis inclusive of a part of the corona, the inner side of the prepuce is used to reconstruct the labia minora, and the penile shaft is inverted to line the vaginal cavity. MATERIAL AND METHODS: A long-term sensitivity evaluation (performed by the Semmes-Weinstein monofilament and the Vibration tests) of 27 reconstructed phalli and 30 clitorises has been performed. RESULTS: The average pressure and vibratory thresholds values for the phallus tip were, respectively, 11.1 g/mm and 3 microm. These values have been compared with the ones of the forearm (donor site). The average pressure and vibratory thresholds values for the clitoris were, respectively, 11.1 g/mm and 0.5 microm. These values have been compared with the ones of the normal male glans, taken from the literature. We also asked the examined patients if they experienced orgasm after surgery, during any sexual practice (ie, we considered only patients who attempted to have orgasm): all female-to-male and 85% of the male-to-female patients reported orgasm. CONCLUSION: With our techniques, the reconstructed genitalia obtain tactile and erogenous sensitivity. To obtain a good tactile sensitivity in the reconstructed phallus, we believe that the coaptation of the cutaneous nerves of the flap with the ilioinguinalis nerve and with one of the 2 nerves of the clitoris is essential in obtaining this result. To obtain orgasm after phalloplasty, we believe that preservation of the clitoris beneath the reconstructed phallus and some preservation of the clitoris hood are essential. To obtain orgasm after a vaginoplasty, the reconstruction of the clitoris from the neurovascular pedicled glans flap is essential. PMID- 17413888 TI - A comparative study of maxillary growth following rotation-advancement and triangular flap unilateral cleft lip repairs: an experimental study in rabbits. AB - The present study was designed to investigate 2 features of maxillary growth following Millard rotation-advancement and Tennison triangular flap cleft lip repairs in rabbits with surgically created defects simulating unilateral cleft lip and cleft alveolus. Sixty purebred New Zealand rabbits were used in this experiment. The animals were divided into 4 groups: 2 control groups (unoperated and unrepaired) and 2 experimental groups (lip repair with rotation-advancement and lip repair with triangular flap). Nineteen metric cranial variables were measured directly from the cleaned skulls. Direct cephalometric measurements were taken in the following dimensions: maxillary length, width and height, posterior facial width, and nasal deflection. Significant differences were noted between Millard rotation-advancement group and Tennison triangular flap group in 2 of 6 measurements of maxillary length. Analysis of variance revealed significantly statistical differences between Millard rotation-advancement group and Tennison triangular flap group in 3 of 6 measures of maxillary width. Analysis of variance revealed no significantly statistical differences between Millard rotation advancement group and Tennison triangular flap group in maxillary height. Statistical differences were noted between Millard rotation-advancement group and Tennison triangular flap group in posterior facial width. There was no significantly statistical difference between Millard rotation-advancement group and Tennison triangular flap group for the nasal deflection measurements. The results of this study indicated that the features of maxillary growth were different between the 2 lip-repair techniques in rabbits with surgically created defects simulating unilateral cleft lip and alveolus. PMID- 17413889 TI - Angiotensin II captopril cotreatment augments angiogenesis in abdominal skin flap in rats. AB - The effect of captopril, angiotensin-converting enzyme inhibitor, on angiogenesis in several reports remained unclear. Its effect on neovascularization in rat abdominal skin flaps was investigated. Flap elevation, based on the right superficial inferior epigastric artery was performed with or without the administration of captopril (10 mg/kg/d), Ang II (100 microg/kg/d), or captopril and Ang II cotreatment. Mean arterial pressure (MAP), microangiography, capillary density measurement, necrosis area determination, laser Doppler flowmetry (LDF), AT1 and vascular endothelial growth factor (VEGF) immunostaining were used to evaluate the effects of captopril and the interaction between captopril and Ang II on the angiogenesis. Ang II and captopril cotreatment improved angiogenesis more than Ang II or captopril alone. The reduction of necrosis, enhancement of vascular network formation, capillary density, VEGF immunostaining, and local blood flow were evident in the cotreated group. We suggest that Ang II and captopril cotreatment improves ischemia-induced angiogenesis and increased viability and vascularity of skin flap in rats. PMID- 17413890 TI - Effects of topical glutathione treatment in rat ischemic wound model. AB - Oxidative stress secondary to ischemia can cause physiopathologic changes that adversely affect wound healing. In this experimental study, we hypothesized that the topical use of esterified glutathione, a well-known antioxidant, can minimize the effects of oxidative stress by an increase in intracellular glutathione and accelerate wound healing by increasing the contraction capacity of fibroblasts and preventing keratinocytes from apoptosis in a rat ischemic wound model. Experimental models were divided into 3 groups as treatment, control, and healthy. Bipedicled flaps were elevated from the dorsum of the rats, and 6-mm punch wounds were created at the end of the first day when the ischemia is most apparent. Wounds were followed histopathologically and immunohistochemically, and matrix metalloproteinase (MMP)-1 and tissue inhibitors of metalloproteinase (TIMP 1) levels were measured by ELISA. Samples were collected at 0, 5, 8, 10, and 12 days. Histopathologic evaluation revealed significant extracellular matrix deposition and reepithelization every fifth day in treatment and healthy groups when compared with control group. Immunohistochemical evaluation revealed increased apoptosis in basal keratinocytes in the control group when compared with the other groups. The evaluation of the samples collected at 5 and 8 days revealed increased MMP-1 levels in treatment and control groups, but the increase in TIMP-1 levels was more significant than MMP-1 levels in treatment group. MMP 1/TIMP-1 ratio was significantly low in the treatment group.Our results showed that topical GSH treatment can reduce oxidative stress, and the reestablishment of the MMP-1/TIMP-1 ratio gives way to adequate and regular extracellular matrix production and reepithelization. It is concluded that esterified GSH, which is experimentally shown to be effective in ischemic wound healing, can be used clinically in ischemic wounds. PMID- 17413891 TI - Dose-dependent effects of a nitric oxide donor in a rat flap model. AB - Previous studies have proven the effectiveness of nitric oxide (NO) donors to enhance flap survival in experimental models. The purpose of this study was to determine the ideal dose of the NO donor spermine/nitric oxide complex (Sper/NO) with respect to flap survival and hemodynamic side effects. Additionally, the influence of the type of application (systemically versus intra-arterial into the flap artery) was observed.Seventy-two male Wistar rats were divided into 9 experimental groups. An extended epigastric adipocutaneous flap (6 x 10 cm) based on the left superficial epigastric artery and vein was raised in each animal. The average percentage of flap necrosis was 69.8% in the control group with ischemia and 29.8% in the non ischemic controls. The average necrosis areas in the 3 groups after preischemic intravenous (i.v.) application of Sper/NO (250, 500, and 750 nmol/kg body weight) were 63.5%, 33.8%, and 38.4%, respectively. The application of similar doses into the flap artery resulted in 63.5%, 72.3%, and 64.3% flap necrosis; 52.7% average flap loss was observed in an additional group receiving Sper/NO adjusted to flap weight (500 nmol/kg flap weight). Only the intravenous application of 500 and 750 nmol/kg resulted in a significant reduction of flap necrosis compared with the ischemic controls (P < 0.01). The drop in mean arterial pressure was less after i.v. application of 500 nmol/kg Sper/NO compared with 750 nmol. Our data show that the preischemic intravenous application of 500 nmol/kg Sper/NO achieved the best results with acceptable side effects. A dose of 250 nmol/kg i.v., as well as the application of Sper/NO into the flap artery, was demonstrated to be ineffective. PMID- 17413892 TI - Acute radial artery ischemia following therapeutic steroid injection. AB - Intra-arterial corticosteroid injection at the wrist can have devastating consequences. Anatomic arterial variation of the hand can lead to atypical presentation. A careful history and examination, with appropriate investigations, can lead to an accurate diagnosis in such difficult cases. PMID- 17413893 TI - Alone in my own hospital. PMID- 17413894 TI - Response to Scalpels and Sparkplugs: the future of plastic surgery. PMID- 17413895 TI - Sentinel node biopsy using lymphoscintigraphy may be complicated by gamma signal from a full bladder. PMID- 17413896 TI - Point of technique: an alternative to the bolus tie-over dressing for full thickness skin grafts. PMID- 17413898 TI - Can we get there if we don't know where we're going? PMID- 17413899 TI - Every breath you take, we'll be watching you. PMID- 17413900 TI - Identifying and learning from mistakes. PMID- 17413901 TI - Imaging inflammation in acute lung injury. PMID- 17413902 TI - On-line monitoring of end-tidal propofol concentration in anesthetized patients. AB - BACKGROUND: Propofol (2,6-diisopropylphenol) has some volatility, so it can be detected in expired breath of individuals receiving intravenous propofol. This study measured volatile propofol exhaled by patients and investigated the relation between exhaled and plasma propofol concentrations. METHODS: Nineteen patients with American Society of Anesthesiologists physical status I or II who were undergoing elective surgery participated in this two-part study. In study 1 (n = 11), anesthesia was induced with 2 mg/kg propofol, 0.1 mg/kg vecuronium, and 2 microg/kg fentanyl. After intubation, propofol was administered continuously for 60 min at each of three rates: 3, 6, and 9 mg x kg(-1) x h(-1). Blood samples were obtained just before each change in the infusion rate, and the plasma concentrations of propofol were measured. The exhaled propofol concentration was measured continuously by means of proton transfer mass spectrometry. End-tidal propofol concentrations during blood sampling were averaged and compared with plasma propofol concentrations. In study 2 (n = 8), after induction of anesthesia, patients received a bolus injection of 2 mg/kg propofol, and the exhaled propofol concentration was measured. RESULTS: Volatile propofol was detected in expired gas from all study patients. From study 1, the authors obtained 24 paired data points, i.e., concentrations of end-tidal and plasma propofol. With Bland-Altman analysis, bias +/- precision was 5.2 +/- 10.4 with 95% limits of agreement of -15.1 and 25.6. In study 2, the exhaled propofol concentration curve showed an obvious peak in all patients. CONCLUSIONS: Agreement between plasma and exhaled propofol concentrations suggests that proton transfer mass spectrometry can be used for real-time propofol monitoring. PMID- 17413903 TI - Real-time monitoring of propofol in expired air in humans undergoing total intravenous anesthesia. AB - BACKGROUND: The physicochemical properties of propofol could allow diffusion across the alveolocapillary membrane and a measurable degree of pulmonary propofol elimination. The authors tested this hypothesis and showed that propofol can be quantified in expiratory air and that propofol breath concentrations reflect blood concentrations. This could allow real-time monitoring of relative changes in the propofol concentration in arterial blood during total intravenous anesthesia. METHODS: The authors measured gas-phase propofol using a mass spectrometry system based on ion-molecule reactions coupled with quadrupole mass spectrometry which provides a highly sensitive method for on-line and off-line measurements of organic and inorganic compounds in gases. In a first sequence of experiments, the authors sampled blood from neurosurgery patients undergoing total intravenous anesthesia and performed propofol headspace determination above the blood sample using an auto-sampler connected to the mass spectrometry system. In a second set of experiments, the mass spectrometry system was connected directly to neurosurgery patients undergoing target-controlled infusion via a T piece inserted between the endotracheal tube and the Y connector of the anesthesia machine, and end-expiratory propofol concentrations were measured on line. RESULTS: A close correlation between propofol whole blood concentration and propofol headspace was found (range of Pearson r, 0.846-0.957; P < 0.01; n = 6). End-expiratory propofol signals mirrored whole blood values with close intraindividual correlations between both parameters (range of Pearson r, 0.784 0.985; n = 11). CONCLUSION: Ion-molecule reaction mass spectrometry may allow the continuous and noninvasive monitoring of expiratory propofol levels in patients undergoing general anesthesia. PMID- 17413904 TI - Analysis of deaths related to anesthesia in the period 1996-2004 from closed claims registered by the Danish Patient Insurance Association. AB - BACKGROUND: Anesthesia is associated with complications, and some of them may be fatal. The authors investigated the circumstances under which deaths were associated with anesthesia. In Denmark, the specialty anesthesiology encompasses emergency medicine, chronic and acute pain medicine, anesthetic procedures, perioperative care medicine, and intensive care medicine. METHODS: The authors retrospectively investigated anesthesia related deaths registered by the Danish Patient Insurance Association. RESULTS: From 1996 to 2004, 27,971 claims were made by the Danish Patient Insurance Association covering all medical specialties, of which 1,256 files (4.5%) were related to anesthesia. In 24 cases, the patient's death was considered to result from the anesthetic procedure: 4 deaths were related to airway management, 2 to ventilation management, 4 to central venous catheter placement, 4 as a result of medication errors, 4 from infusion pump problems, and 4 after complications from regional blockades. Severe hemorrhage caused 1 death, and in 1 case the cause was uncertain. CONCLUSIONS: Several of the 24 deaths could potentially have been avoided by more extended use of airway algorithm, thorough preoperative evaluation, training, education, and use of protocols for diagnosis and treatment. PMID- 17413905 TI - Factor V Leiden does not affect bleeding in aprotinin recipients after cardiopulmonary bypass. AB - BACKGROUND: Carriers of the factor V Leiden mutation (FVL) are resistant to activated protein C proteolysis. Therefore, they are at increased risk of thromboembolic events. Aprotinin is an unspecific proteinase inhibitor frequently used during cardiac surgery procedures to reduce bleeding. However, aprotinin may cause thromboembolic complications after cardiopulmonary bypass (CPB). The primary endpoint of this study was the amount of blood loss after CPB in aprotinin recipients, and secondary endpoints were thromboembolic complications. METHODS: A total of 1,447 consecutive patients who underwent cardiac surgery with CPB were prospectively enrolled. All patients were screened for FVL by a fluorescence-based polymerase chain reaction method. Linear and logistic regression analyses were performed to assess associations of FVL on bleeding and thromboembolic complications. RESULTS: One hundred seven individuals (7.4%) were heterozygous FVL carriers. No difference was found between FVL carriers and noncarriers regarding age, sex, CPB, type of operation, EuroSCORE, antiplatelet treatment, and reoperation. FVL was not significantly associated with postoperative blood loss, whereas a significant influence was found for female sex (P < 0.0001), duration of CPB (P < 0.0001), reoperation (P = 0.001), and preoperative antiplatelet treatment (P < 0.002). Multiple linear regression analysis for total blood loss had an observed power of at least 99%. FVL carriers faced the same risk for postoperative transfusion (P = 0.391), reoperation (P = 0.675), myocardial infarction (P = 0.44), stroke (P = 0.701), and 30-day mortality (P = 0.4) as did noncarriers. CONCLUSIONS: These data suggest that FVL carriers do not have reduced blood loss compared with noncarriers. Furthermore, the combination of aprotinin and FVL does not enhance the risk for thromboembolic complications. PMID- 17413906 TI - Current practices in sedation and analgesia for mechanically ventilated critically ill patients: a prospective multicenter patient-based study. AB - BACKGROUND: The authors conducted a patient-based survey of practices to fully describe the assessment and the management of pain and sedation of a large cohort of mechanically ventilated patients during their first week of intensive care unit (ICU) stay. METHODS: A total of 1,381 adult patients were included in a prospective, observational study in 44 ICUs in France. Pain and sedation assessment, analgesic and sedative use, and analgesic management during procedural pain were collected on days 2, 4, and 6 of the ICU stay. RESULTS: The observed rates of assessment on day 2 for sedation (43%) and analgesia (42%) were significantly smaller than that of use of sedatives (72%) and opioids (90%), also noted on days 4 and 6. The use of protocols/guidelines for sedation/analgesia in the ICU reduced the proportion of patients who were treated, although not evaluated. A large proportion of assessed patients were in a deep state of sedation (40-50%). Minor changes in the dosages of the main prescribed agents for sedation (midazolam, propofol) and analgesia (sufentanil, fentanyl, morphine, remifentanil) were found across 6 days of the patient's ICU stay. Procedural pain was specifically managed for less than 25% of patients; during those procedures, the proportion of patients with pain significantly increased from the baseline pain evaluation. CONCLUSIONS: Excessively deep states of sedation and a lack of analgesia during painful procedures must be prevented. To facilitate systematic pain and sedation assessment and to adjust daily drug dosages accordingly, it seems crucial to promote educational programs and elaboration of protocols/guidelines in the ICU. PMID- 17413907 TI - Behavior of entropy/complexity measures of the electroencephalogram during propofol-induced sedation: dose-dependent effects of remifentanil. AB - BACKGROUND: Several new measures based on the regularity of the electroencephalogram signal for the assessment of depth of anesthesia/sedation have been proposed recently. In this study we analyze the influence of remifentanil and electroencephalogram frequency content of the performance of a set of such measures. METHODS: Forty-five patients with American Society of Anesthesiologists physical status I were randomly allocated to one of three groups according to the received dose of predicted effect compartment-controlled remifentanil (0, 2, and 4 ng/ml). All 45 patients received stepwise increased effect site concentration-controlled dose of propofol. At every step of propofol increase, the Observer's Assessment of Alertness/Sedation score was assessed. The following measures were calculated from the electroencephalographic signal: spectral entropy, approximate entropy, Higuchi fractal dimension, Lempel-Ziv complexity, relative beta ratio, and SyncFastSlow measure. RESULTS: The behavior of the electroencephalogram-based measures is highly sensitive to the frequency content of the signal and the dose of remifentanil. The prediction probability with respect to the Observer's Assessment of Alertness/Sedation score of the most discriminative measure, the Higuchi fractal dimension, dropped from 0.90 (electroencephalographic frequency band 6-47 Hz, no remifentanil) to 0.55 when the frequency band was changed to 0.5-19 Hz and to 0.83 when remifentanil concentration was increased to 4 ng/ml. The coeffect of remifentanil on electroencephalographic regularity is bimodal depending on the frequency band of the signal. CONCLUSIONS: Cutting off high frequencies from the electroencephalogram and increased remifentanil concentration deteriorate the performance of the electroencephalogram-based entropy/complexity measures as indicators of the depth of propofol sedation. PMID- 17413908 TI - Identification of sensory blockade by somatosensory and pain-induced evoked potentials. AB - BACKGROUND: To date, the anesthesia-induced blockade of nociceptive inputs is insufficiently reflected by commercially available electroencephalographic depth of-anesthesia monitors. The aim of the current study was to evaluate the potential of somatosensory (SSEP) and intracutaneous pain evoked (iSEP) potentials during remifentanil and propofol anesthesia as electroencephalographic indicators of the nociceptive blockade. METHODS: Ten healthy men were investigated in a double-blind crossover design during three sessions with remifentanil, propofol, and placebo administration. All dosages were increased in a step-by-step mode. SSEP and iSEP recordings were performed followed by subjective pain ratings and measurement of level of sedation (modified Observer's Assessment of Alertness and Sedation Scale). Changes from baseline in evoked potential components, pain ratings, and sedation scale were assessed by Bonferroni-Holms-corrected Wilcoxon tests. RESULTS: Pain ratings were significantly reduced by remifentanil. Sedation scale was significantly reduced by propofol. Early SSEP components were not affected by medication. The amplitudes of the long latency SSEP components increased significantly with remifentanil, decreased with propofol, and did not change with placebo. The amplitudes of long latency components of the iSEP decreased significantly with both remifentanil and propofol and did not change with placebo. CONCLUSION: Long latency components of the SSEP are differently affected by remifentanil and propofol administration. Further studies are needed to clarify whether they can serve as a specific indicator of the nociceptive blockade during anesthesia. PMID- 17413909 TI - Oxygen tension modulates inhibition of L-type calcium currents by isoflurane in human atrial cardiomyocytes. AB - BACKGROUND: Myocardial L-type Ca(2+) currents (I(Ca,L)) are inhibited by isoflurane in the presence of a partial pressure of oxygen (P(O2)) of 150 mmHg. In guinea pig cardiomyocytes, I(Ca,L) are inhibited by reduced oxygen tensions. The authors therefore analyzed the effects of P(O2) on I(Ca,L) in human cardiomyocytes and the effects of isoflurane at reduced P(O2). METHODS: Atrial cardiomyocytes were prepared from specimens of patients undergoing open-heart surgery and superfused with either a high or a low P(O2) (150 or 12 +/- 1 mmHg) while I(Ca,L) were measured with the whole cell patch clamp technique. RESULTS: Basal I(Ca,L) were not changed by the P(O2) (range, 9-150 mmHg) at 21 degrees or 36 degrees C. The reducing agent 1,4-dithiothreitol (DTT) left I(Ca,L) unaffected, and the oxidizing agent 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) irreversibly inhibited I(Ca,L). The P(O2) significantly affected the inhibition of I(Ca,L) by isoflurane (1 minimum alveolar concentration) that decreased I(Ca,L) by 17 +/- 2.0% at the high P(O2) but only by 5.8 +/- 2.9% (P = 0.037) at the low P(O2). The inhibition of I(Ca,L) by isoflurane was also significantly diminished (P = 0.018) by a low P(O2) when isoflurane effects at both P(O2) conditions were compared in the same cell. CONCLUSIONS: In contrast to the situation in guinea pigs, basal I(Ca,L) in human atrial cardiomyocytes was not sensitive to acute P(O2) changes over a wide range. This might be explained by a lack of oxygen-sensitive splice variants of L-type calcium channel subunits. The P(O2), however, has a decisive role for the effects of isoflurane on I(Ca,L). PMID- 17413910 TI - Regional gas exchange and cellular metabolic activity in ventilator-induced lung injury. AB - BACKGROUND: Alveolar overdistension and repetitive derecruitment-recruitment contribute to ventilator-induced lung injury (VILI). The authors investigated (1) whether inflammatory cell activation due to VILI was assessable by positron emission tomography and (2) whether cell activation due to dynamic overdistension alone was detectable when other manifestations of VILI were not yet evident. METHODS: The authors assessed cellular metabolic activity with [(18)F]fluorodeoxyglucose and regional gas exchange with [(13)N]nitrogen. In 12 sheep, the left ("test") lung was overdistended with end-inspiratory pressure of 50 cm H(2)O for 90 min, while end-expiratory derecruitment of this lung was either promoted with end-expiratory pressure of -10 cm H(2)O in 6 of these sheep (negative end-expiratory pressure [NEEP] group) or prevented with +10 cm H(2)O in the other 6 (positive end-expiratory pressure [PEEP] group) to isolate the effect of overdistension. The right ("control") lung was protected from VILI. RESULTS: Aeration decreased and shunt fraction increased in the test lung of the NEEP group. [(18)F]fluorodeoxyglucose uptake of this lung was higher than that of the control lung and of the test lung of the PEEP group, and correlated with neutrophil count. When normalized by tissue fraction to account for increased aeration of the test lung in the PEEP group, [(18)F]fluorodeoxyglucose uptake was elevated also in this group, despite the fact that gas exchange had not yet deteriorated after 90 min of overdistension alone. CONCLUSION: The authors could detect regional neutrophil activation in VILI even when end-expiratory derecruitment was prevented and impairment of gas exchange was not evident. Concomitant end-expiratory derecruitment converted this activation into profound inflammation with decreased aeration and regional shunting. PMID- 17413911 TI - Isoflurane depresses the response of inspiratory hypoglossal motoneurons to serotonin in vivo. AB - BACKGROUND: Endogenous serotonin (5-HT) provides important excitatory drive to inspiratory hypoglossal motoneurons (IHMNs). In vitro studies show that activation of postsynaptic 5-HT receptors decreases a leak K+ channel conductance and depolarizes hypoglossal motoneurons (HMNs). In contrast, volatile anesthetics increase this leak K+ channel conductance, which causes neuronal membrane hyperpolarization and depresses HMN excitability. Clinical studies show upper airway obstruction, indicating HMN depression, even at subanesthetic concentrations. The authors hypothesized that if anesthetic activation of leak K+ channels caused neuronal depression in vivo, this effect could be antagonized with serotonin. In this case, the neuronal response to picoejected serotonin would be greater during isoflurane than with no isoflurane. METHODS: Studies were performed in decerebrate, vagotomized, paralyzed, and mechanically ventilated dogs during hypercapnic hyperoxia. The authors studied the effect of approximately 0.3 minimum alveolar concentration (MAC) isoflurane on the spontaneous discharge frequency patterns of single IHMNs and on the neuronal response to picoejection of 5-HT. RESULTS: Normalized data (mean +/- SD, n = 19) confirmed that 0.3 +/- 0.1 MAC isoflurane markedly reduced the spontaneous peak discharge frequency by 48 +/- 19% (P < 0.001) and depressed the slope of the spontaneous discharge patterns. The increase in neuronal frequency in response to 5-HT was reduced by 34 +/- 22% by isoflurane (P < 0.001). CONCLUSION: Subanesthetic concentrations of isoflurane strongly depressed canine IHMNs in vivo. The neuronal response to 5-HT was also depressed by isoflurane, suggesting that anesthetic activation of leak K+ channels, which is expected to result in a larger 5-HT response, was not a dominant mechanism in this depression. PMID- 17413912 TI - Xenon mitigates isoflurane-induced neuronal apoptosis in the developing rodent brain. AB - BACKGROUND: Anesthetics, including isoflurane and nitrous oxide, an antagonist of the N-methyl-D-aspartate subtype of the glutamate receptor, have been demonstrated to induce apoptotic neurodegeneration when administered during neurodevelopment. Xenon, also an N-methyl-D-aspartate antagonist, not only lacks the characteristic toxicity produced by other N-methyl-D-aspartate antagonists, but also attenuates the neurotoxicity produced by this class of agent. Therefore, the current study sought to investigate xenon's putative protective properties against anesthetic-induced neuronal apoptosis. METHOD: Separate cohorts (n = 5 or 6 per group) of 7-day-old rats were randomly assigned and exposed to eight gas mixtures: air, 75% nitrous oxide, 75% xenon, 0.75% isoflurane, 0.75% isoflurane plus 35% or 75% nitrous oxide, 0.75% isoflurane plus 30% or 60% xenon for 6 h. Rats were killed, and cortical and hippocampal apoptosis was assessed using caspase-3 immunostaining. In separate cohorts, cortices were isolated for immunoblotting of caspase 3, caspase 8, caspase 9, and cytochrome c. Organotypic hippocampal slices of postnatal mice pups were derived and cultured for 24 h before similar gas exposures, as above, and subsequently processed for caspase-3 immunostaining. RESULTS: In vivo administration of isoflurane enhances neuronal apoptosis. When combined with isoflurane, nitrous oxide significantly increases whereas xenon significantly reduces apoptosis to a value no different from that of controls. In vitro studies corroborate the ability of xenon to attenuate isoflurane-induced apoptosis. Isoflurane enhanced expression of indicators of the intrinsic and common apoptotic pathways; this enhancement was increased by nitrous oxide but attenuated by xenon. CONCLUSIONS: The current study demonstrates that xenon prevents isoflurane-induced neonatal neuronal apoptosis. PMID- 17413913 TI - Subhypnotic doses of isoflurane impair auditory discrimination in rats. AB - BACKGROUND: Isoflurane at subhypnotic doses is known to affect cellular and network activity in the auditory pathway, but the behavioral effects of these concentrations of isoflurane on auditory processing have not been tested previously. The authors tested the hypothesis that subhypnotic doses of isoflurane would impair auditory discrimination in rats. METHODS: Rats were tested on their ability to discriminate up versus down frequency-modulated sweeps using three different pairs of sweeps ("Long," "Med," "Short"), whose frequency range and duration were varied systematically to make the discrimination more difficult. Rats were tested daily in the absence and presence of isoflurane at 0.2% or 0.4%. The effects of isoflurane (0%, 0.2%, and 0.4%) on performance (= % correct) and efficiency (= time/trial) were assessed using regression analysis. RESULTS: The effect of isoflurane was stimulus-dependent: performance for the Long stimulus pair was unaffected by isoflurane, performance on the Med stimulus pair was impaired only by 0.4% isoflurane, and performance on the Short stimulus pair was impaired by both 0.2% and 0.4% isoflurane. In contrast, isoflurane decreased efficiency equally for all stimulus pairs at 0.4% and had no effect at 0.2%. CONCLUSIONS: The stimulus dependence of the effect of isoflurane on performance suggests that it is unlikely this effect was secondary to effects on memory, motivation, or motor function. These data indicate that doses of isoflurane known to produce modest effects on neural responses alter cortical sensory processing. PMID- 17413914 TI - Neuromuscular blocking agents' differential bronchoconstrictive potential in Guinea pig airways. AB - BACKGROUND: Neuromuscular blocking agents are designed to antagonize nicotinic cholinergic receptors on skeletal muscle but also antagonize muscarinic receptors. Several muscle relaxants have the potential to promote bronchoconstriction due to unintended effects exemplified by histamine release of atracurium or mivacurium and detrimental interactions with muscarinic receptors by rapacuronium. Although interactions of muscle relaxants with muscarinic receptors have been extensively characterized in vitro, limited information is available on their potential interactions with airway tone in vivo. METHODS: Changes in pulmonary inflation pressures and heart rates induced by vagal nerve stimulation and intravenous acetylcholine were measured in the absence and presence of increasing doses of gallamine, pancuronium, mivacurium, vecuronium, cisatracurium, rocuronium, or rapacuronium in guinea pigs. Mivacurium's and rapacuronium's potential of inducing bronchoconstriction by histamine release was also evaluated. RESULTS: Rapacuronium potentiated both vagal nerve-stimulated and intravenous acetylcholine-induced increases in airway pressures, which were totally blocked by atropine but not pyrilamine. Vecuronium, rocuronium, mivacurium, and cisatracurium were devoid of significant airway effects. Mivacurium, at high doses, increased pulmonary inflation pressures, which were attenuated by pyrilamine. CONCLUSION: Rapacuronium was unique among muscle relaxants evaluated in that it potentiated both vagal nerve- and intravenous acetylcholine-induced bronchoconstriction with no evidence of histamine release. The dual detrimental interactions of rapacuronium with muscarinic receptors previously demonstrated in vitro correlate with in vivo muscarinic receptor mechanisms of bronchoconstriction and may account for the profound bronchoconstriction seen with its clinical use. These findings may establish pharmacologic characteristics to avoid with new muscle relaxants intended for clinical use. PMID- 17413915 TI - Fascia iliaca compartment blockade for acute pain control in hip fracture patients: a randomized, placebo-controlled trial. AB - BACKGROUND: Hip fracture patients are in severe pain upon arrival at the emergency department. Pain treatment is traditionally based on systemic opioids. No study has examined the effect of fascia iliaca compartment blockade (FICB) in acute hip fracture pain management within a double-blind, randomized setup. METHODS: Forty-eight patients with suspected hip fracture were included immediately after arrival in the emergency department, before x-ray confirmation of their fracture. Included patients were randomly assigned to two groups of 24. In the FICB group, the patients received an FICB with 1.0% mepivacaine and a placebo intramuscular injection of isotonic saline. In the morphine group, the patients received a placebo FICB with 0.9% saline and an intramuscular injection of 0.1 mg/kg morphine. Patients received intravenous rescue morphine when necessary. RESULTS: Maximum pain relief was superior in the FICB group both at rest (P < 0.01) and on movement (P = 0.02). The median total morphine consumption was 0 mg (interquartile range, 0-0 mg) in the FICB group and 6 mg (interquartile range, 5-7 mg) in the morphine group (P < 0.01). More patients (P = 0.05) were sedated in the morphine group at 180 min after block placement as compared with the FICB group. CONCLUSION: Pain relief was superior at all times and at all measurements in the FICB group. The study supports the use of FICB in acute management of hip fracture pain because it is an effective, easily learned procedure that also may reduce opioid side effects in this fragile, elderly group of patients. PMID- 17413916 TI - Magnetic resonance imaging in patients with spinal neurostimulation systems. AB - BACKGROUND: The safety of performing magnetic resonance imaging (MRI) in patients with spinal cord stimulation (SCS) systems needs to be documented. A prospective in vivo study in patients with SCS, exploring the changes produced by MRI and the associated side effects, was performed. METHODS: After ethics committee approval and patient consent, 31 consecutive patients with SCS at different spinal levels requiring a scheduled MRI evaluation were studied during an 18-month period. All MRIs were performed with a 1.5-T clinical use magnet and a specific absorption rate of no more than 0.9 W/kg. Frequency tables were used for the descriptive study, whereas comparative evaluations were made with the chi-square test for qualitative variables and single-factor analysis of variance for quantitative variables. RESULTS: The mean patient age was 49 +/- 9.5 yr; 67.7% were women (n = 21), and 32.3% were men (n = 10). None of the patients experienced hemodynamic, respiratory, or neurologic alterations. Reported changes were as follows: increased temperature in the generator's area (n = 2, 6.5%); increased in the intensity of the stimulation (n = 1, 3.2%); impedance greater than 4,000 Omega on several of the electrodes in the leads (n = 1, 3.2%); telemetry not possible (n = 2, 6.5%). Radiologic evaluation after MRI revealed no spatial displacements of the SCS leads in any case. CONCLUSION: Under the conditions of the described protocol, MRI in patients with SCS systems resulted in few complications. None of the recorded problems were serious, and in no case were patients harmed or the systems reprogrammed. Maximum patient satisfaction was reported in all cases. PMID- 17413917 TI - Spinal cannabinoid receptor type 2 activation reduces hypersensitivity and spinal cord glial activation after paw incision. AB - BACKGROUND: Cannabinoids bind to cannabinoid receptors type 1 and 2 and produce analgesia in several pain models, but central side effects from cannabinoid 1 receptors limit their clinical use. Cannabinoid 2 receptors reduce inflammatory responses in the periphery by acting on immune cells, and they are present on glia in the central nervous system. This study tested whether spinal cannabinoid activation would induce analgesia, glial inhibition, and central side effects in a postoperative model or incisional pain. METHODS: Rats underwent paw incision surgery, with intrathecal injections of cannabinoid agonists and antagonists and assessment of withdrawal thresholds and behavioral side effects. Spinal glial activation was determined by immunohistochemistry. RESULTS: Intrathecal administration CP55940 reduced postoperative hypersensitivity (91 +/- 9% maximum possible effect; P < 0.05), and this was prevented by intrathecal administration of both cannabinoid 1 receptor (AM281) and cannabinoid 2 receptor (AM630) antagonists. CP55940 also caused several behavioral side effects, and these were prevented by the cannabinoid 1 receptor but not by the cannabinoid 2 receptor antagonist. Intrathecal injection of the cannabinoid 2 receptor agonist JWH015 reversed postoperative hypersensitivity (89 +/- 5% maximum possible effect; P < 0.05), and this was reversed by the cannabinoid 2 but not by the cannabinoid 1 receptor antagonist. JWH015, which did not induce behavioral side effects, reduced paw incision induced microglial and astrocytic activation in spinal cord (P < 0.05). CONCLUSIONS: These data indicate that intrathecal administration of cannabinoid receptor agonists may provide postoperative analgesia while reducing spinal glial activation, and that selective cannabinoid 2 receptor agonists may do so without central side effects. PMID- 17413918 TI - Spinal prostaglandins facilitate exaggerated A- and C-fiber-mediated reflex responses and are critical to the development of allodynia early after L5-L6 spinal nerve ligation. AB - BACKGROUND: Spinal prostaglandins are important in the early pathogenesis of spinal nerve ligation (SNL)-induced allodynia. This study examined the effect of SNL on the expression of cyclooxygenase (COX)-1, COX-2, and prostaglandin E2 receptors in the rat lumbar spinal cord, and the temporal and pharmacologic relation of these changes to the exaggerated A- and C-fiber-mediated reflex responses and allodynia, 24 h after injury. METHODS: Male Sprague-Dawley rats, fitted with intrathecal catheters, underwent SNL or sham surgery. Paw withdrawal threshold, electromyographic analysis of the biceps femoris flexor reflex, and immunoblotting of the spinal cord were used. RESULTS: Both allodynia (paw withdrawal threshold of < or = 4 g) and exaggerated A- and C-fiber-mediated reflex responses (i.e., decrease in activation threshold, increase in evoked activity, including windup; P < 0.05) were evident 24 h after SNL but not sham surgery. Allodynic animals exhibited significant increases in prostaglandin E2 receptor (subtypes 1-3) and COX-1 (but not COX-2) expression in the ipsilateral lumbar dorsal horn. The corresponding ventral horns and contralateral dorsal horn were unchanged from sham controls. Exaggerated A- and C-fiber-mediated reflex responses were significantly attenuated by intrathecal SC-560 or SC-51322, but not SC-236, given 24 h after SNL. CONCLUSION: These results provide further evidence that spinal prostaglandins, derived primarily from COX-1, are critical in the exaggeration of A- and C-fiber input and allodynia, 24 h after SNL. PMID- 17413919 TI - Gender differences in anesthesiologists' annual incomes. AB - BACKGROUND: Specialty, work effort, and gender have been shown to be associated with physicians' annual incomes; however, careful examination of the association between provider gender and annual income after correcting for other factors likely to influence income has not been conducted for anesthesiologists. METHODS: Survey responses collected throughout the 1990s from 819 actively practicing anesthesiologists and linear regression analysis were used to determine the association between provider gender and annual incomes after controlling for work effort, provider characteristics, and practice characteristics. RESULTS: White female anesthesiologists reported working 12% fewer annual hours than their white male counterparts. White female anesthesiologists had practiced medicine for fewer years than white males and were more likely to be employees, as opposed to having an ownership interest in the practice, but less likely to be board certified. After adjustment for work effort, provider characteristics, and practice characteristics, white females' mean annual income was 236,628 dollars, or 60,337 dollars (20%) lower than that for white males (95% confidence interval, 81,674 dollars lower to 39,001 dollars lower; P < 0.001). CONCLUSIONS: During the 1990s, female gender was associated with lower annual incomes among anesthesiologists. These findings warrant further exploration to determine what factors might cause these gender-based income differences. PMID- 17413920 TI - Assessment of competency in anesthesiology. AB - Assessment of competency in traditional graduate medical education has been based on observation of clinical care and classroom teaching. In anesthesiology, this has been relatively easy because of the high volume of care provided by residents under the direct observation of faculty in the operating room. With the movement to create accountability for graduate medical education, there is pressure to move toward assessment of competency. The Outcome Project of the Accreditation Council for Graduate Medical Education has mandated that residency programs teach six core competencies, create reliable tools to assess learning of the competencies, and use the data for program improvement. General approaches to assessment and how these approaches fit into the context of anesthesiology are highly relevant for academic physicians. PMID- 17413921 TI - We are what we make: transforming research in anesthesiology: the 45th Rovenstine Lecture. PMID- 17413922 TI - Career National Institutes of Health funding and scholarship of chairpersons of academic departments of anesthesiology and surgery. PMID- 17413923 TI - Practice guidelines for obstetric anesthesia: an updated report by the American Society of Anesthesiologists Task Force on Obstetric Anesthesia. PMID- 17413924 TI - Central sensitization: uncovering the relation between pain and plasticity. PMID- 17413925 TI - Unusual cause of postoperative blindness. PMID- 17413926 TI - Awareness without recall during anesthesia for electroconvulsive therapy. PMID- 17413927 TI - Vascular thrombosis associated with aprotinin and deep hypothermic circulatory arrest: where are we in 2006? PMID- 17413929 TI - Thrombosis after hypothermic circulatory arrest for cardiovascular surgery, antifibrinolytic drugs, and thrombophilia. PMID- 17413930 TI - Association of high tidal volume with postpneumonectomy failure. PMID- 17413932 TI - Therapy in postpuncture meningitis. PMID- 17413933 TI - Gloved and masked-will gowns be next? Let the data (not logic) decide this issue. PMID- 17413934 TI - Think about room air. PMID- 17413935 TI - Parturients expect safe and clean regional anesthesia. PMID- 17413939 TI - Can precise data improve a nonprecise anesthetic? PMID- 17413940 TI - Use of vasopressin bolus and infusion to treat catecholamine-resistant hypotension during pheochromocytoma resection. PMID- 17413942 TI - Impact of terlipressin on hepatosplanchnic perfusion: "only the dose makes a thing not a poison" (paracelsus). PMID- 17413943 TI - GlideScope intubation assisted by fiberoptic scope. PMID- 17413944 TI - Optimizing GlideScope laryngoscopy: an in vitro study on an airway model. PMID- 17413946 TI - Potential research projects. AB - PURPOSE: This review introduces 7 topics for potential research opportunities. METHODS: Unanswered questions over a period of many years regarding ophthalmic issues are listed and summarized. RESULTS: If any or all of the topics are chosen for research, the results will come from individual studies. CONCLUSIONS: If the ideas found are worthy of study, there is potential for great patient benefit. PMID- 17413947 TI - Outcomes of LASIK for myopia with FDA-approved lasers. AB - PURPOSE: To report expected outcomes of laser in situ keratomileusis (LASIK) for myopia and myopic astigmatism from existing US Food and Drug Administration (FDA) data. METHODS: Data from Summaries of Safety and Effectiveness for each of the 12 lasers approved by the FDA for LASIK for myopia or myopic astigmatism between 1998 and 2004 were recorded from the FDA Web site. The Cochran-Armitage test for trend was used to determine whether improvements in outcomes occurred with laser technology changes. RESULTS: For all patients, there was a statistically significant trend toward improvement with improved laser technology in the proportion of patients with uncorrected visual acuity (UCVA) of 20/20 or better, UCVA of 20/40 or better, results within +/-0.50 D of intended correction, results within +/-1.00 D of the intended correction, and night vision symptoms (all P < 0.0002). Because there were preoperative differences across laser types, subgroup analyses were also completed. The results for subgroup analyses (high myopia, low to moderate myopia, spherical myopia, and myopic astigmatism) for visual acuity and refractive error outcomes were similar to results for analyses for all groups combined. Conversely, there was no difference across laser types in the proportion of patients who experienced dry eye symptoms or for the proportion of patients with low to moderate myopia who experienced night vision symptoms that were worse or significantly worse than before LASIK. CONCLUSIONS: LASIK provides excellent visual acuity and refractive error outcomes. Night vision and dryness symptoms still occur in a significant proportion of patients. Future studies should seek to determine whether additional changes in technology, patient selection criteria, or postoperative treatment could reduce or eliminate these symptoms. PMID- 17413948 TI - Nocardia keratitis: species, drug sensitivities, and clinical correlation. AB - PURPOSE: To correlate the clinical presentation and treatment outcome of Nocardia keratitis with the time to diagnosis, different species and with the drug sensitivity pattern. METHOD: Patients with Nocardia corneal ulcers were studied at a tertiary eye care center. Speciation of Nocardia isolates was done by sequencing the 16s rRNA gene. Clinical response to treatment was assessed by chart review. RESULTS: Twenty one (65.3%) patients presented within 15 days of the onset of symptoms with typical clinical features of Nocardia keratitis, ie, a ring-like distribution of superficial infiltrates in a wreath pattern. Eight patients (25%) who presented after 15 days and within 30 days had an ulcer resembling fungal keratitis. N. Cyriacigeorgica (n = 11; 34.37%), N. asteroides (n = 9; 28%), N. farcinica (n = 7; 22%,) and N. Otitidiscaviarum (n = 5; 16%). All the species had 100% sensitivity to amikacin, sulphamethoxazole, imipenem and co-trimoxazole. Time to diagnosis of the infection was significantly associated with the different types of clinical presentation; those presenting early having the typical clinical picture (P = 0.004). Patients (73%) presenting within 15 days showed a highest recovery rate. (P = 0.045). The recovery time of the patients when compared with species showed those who were infected with N. cyriageorgica had a healing time of less than 15 days. Clinically, healing was faster when treated with 2% amikacin. Visual outcome improved in fourteen patients (44%) and sixteen (50%) patients remained the same (P = 0.0001). CONCLUSION: Characteristic clinical picture of Nocardia is dependant on early presentation. It could be mistaken for fungal keratitis. Microbiological confirmation is important. Drug of choice for Nocardia keratitis is amikacin. PMID- 17413949 TI - Effect of oral linoleic and gamma-linolenic acid on meibomian gland dysfunction. AB - PURPOSE: To assess the effect of oral linoleic and gamma-linolenic acid, 2 omega 6 essential fatty acids, on meibomian gland dysfunction (MGD). METHODS: Fifty seven patients with MGD (27 men and 30 women) were randomly divided into 3 groups of 19. Group A received tablets containing linoleic acid (28.5 mg) and gamma linolenic acid (15 mg) once daily. Group B performed eyelid hygiene once daily. Group C received both treatments. Initially and after 60 and 180 days of therapy, all patients completed a self-evaluation questionnaire on ocular surface disorders and underwent slit-lamp examination. The following signs were evaluated: eyelid edema, eyelid margin hyperemia, meibomian secretion appearance, meibomian gland obstruction, foam collection in the tear meniscus, conjunctival hyperemia, conjunctival papillae, and corneal staining. RESULTS: Statistically significant improvement in symptoms occurred in all groups. After 180-day therapy, group A showed significant reduction in secretion turbidity (P = 0.02) and meibomian gland obstruction (P = 0.0001), whereas group B had significant reduction in eyelid edema (P = 0.02), corneal staining (P = 0.01), secretion turbidity (P = 0.01), and meibomian gland obstruction (P = 0.0001). Group C had significant reduction in eyelid edema (P = 0.003), foam collection in the tear meniscus (P = 0.02), corneal staining (P = 0.02), secretion turbidity (P = 0.0001), and meibomian gland obstruction (P = 0.0001). CONCLUSIONS: Therapy with linoleic and gamma-linolenic acid tablets along with eyelid hygiene improves symptoms and reduces eyelid margin inflammation in MGD more than either omega-6 fatty acids or eyelid hygiene alone. PMID- 17413950 TI - White light confocal microscopy of preserved human corneas from an eye bank. AB - PURPOSE: To verify the possibility of using a commercially available ophthalmologic white light confocal microscope for imaging optical sections of donated corneas preserved at 4 degrees C, especially for endothelial evaluation. METHODS: Sixteen corneas donated to the Warsaw Eye Bank but excluded from use in surgery for serological or morphologic reasons were examined using a ConfoScan 3 confocal scanning microscope (Nidek Technologies, Padova, Italy) and a Konan Eye Bank KeratoAnalyzer specular microscope (Konan Medical, Inc.; Hyogo, Japan). Images of corneal structures were obtained, including epithelium, corneal nerves, stroma, and endothelium. The endothelial cell density was calculated for both microscopes, and the results were compared. RESULTS: For images obtained with the specular microscope, mean (+/- SD) endothelial cell density was 2168.6 +/- 404.0 cells/mm; for confocal microscope images, mean +/- SD was 2090.9 +/- 369.1 cells/mm. There was no significant difference between the methods. CONCLUSIONS: White light confocal microscopy can be used for high-magnification imaging of corneas preserved in an eye bank with the 4 degrees C method, and images obtained permit evaluation of endothelium. Although the quality of confocal microscopy images of eye bank corneas is generally lower than that achieved with in vivo examinations, and although technical adaptations are needed for easier and safer application to corneas intended for transplantation, confocal microscopy is a promising new tool for evaluation of collected corneas. PMID- 17413951 TI - In situ versus whole-globe harvesting of corneal tissue from remote donor sites: effects on initial tissue quality. AB - PURPOSE: There are 2 methods of corneal tissue procurement currently in widespread use: in situ extraction of the corneal button directly to preservation media and whole-globe enucleation of eyes with removal of the button to preservation media at a later time. This study evaluates the effects of these 2 procurement procedures on the initial quality of donor corneal tissue. METHODS: Slit-lamp examination results and endothelial cell counts were compared for a total of 468 donor corneas harvested at 2 remote locations: one where in situ procurement was practiced and the other that used whole-globe enucleation procedures. RESULTS: In both univariate and multivariate analysis, in situ corneas were found to have a lower incidence of moderate or severe haze and folds in Descemet membrane. No differences in mean endothelial cell counts were noted between the 2 populations of donated tissue. CONCLUSIONS: In situ procurement of corneal tissue results in higher initial corneal tissue quality than whole-globe procedures. PMID- 17413952 TI - National survey on bullous keratopathy in Japan. AB - PURPOSE: To present the results of a national survey on bullous keratopathy (BK) in Japan. METHODS: A cross-sectional national survey was conducted for 963 eyes with BK seen between 1999 and 2001 by members of the Japan Cornea Society. Demographic characteristics, type of surgery, complications, and postoperative outcome were analyzed. RESULTS: BK accounted for 24.2% (963 eyes) of total keratoplasties performed during the period. Graft clarity was maintained in 77.4% of cases, and immunologic rejection and elevated intraocular pressure was noted in 10.8% and 15.3%, respectively. Cataract surgery was the most common cause of BK (n = 428, 44.4%), and phacoemulsification and aspiration were performed in approximately 40% of cases. BK secondary to laser iridotomy (LI) was the second most common cause of BK (n = 225, 23.4%). LI was performed as a prophylactic measure in approximately one half of these cases. BK developed with a mean duration of 6.8 years after LI. Fuchs dystrophy was the cause of BK in 18 eyes (1.9%). CONCLUSIONS: The causes of BK in Japan are considerably different from those in other Western countries. LI-related BKs showed a remarkably high number, whereas Fuchs dystrophy was observed only rarely. PMID- 17413953 TI - Experience and 12-month results of descemet-stripping endothelial keratoplasty (DSEK) with a small-incision technique. AB - PURPOSE: To report our clinical experience and 12-month results of small-incision Descemet-stripping endothelial keratoplasty (DSEK). METHODS: Prospective study of 11 eyes of 9 patients who had DSEK. The DSEK technique consisted of stripping the Descemet membrane and endothelium from the recipient cornea. The donor button was prepared by manual dissection and inserted through a 5-mm incision. Air, sulfur hexafluoride (SF6), or perfluoropropane (C3F8) was used both at the end of surgery and in subsequent dislocations to promote donor tissue adherence. RESULTS: Mean age was 79.6 years (range, 66-91 years), and minimum follow-up was 12 months (range, 12-18 months). Nine eyes had donor tissue dislocation postoperatively, 8 of which received intervention with either SF6 (n = 4) or C3F8 (n = 4). In 1 patient with repeat dislocation, Tisseel glue in combination with C3F8 was used. Preoperative best-corrected visual acuity (BCVA) was 6/24 or worse in all patients. Postoperatively, 6/11 eyes (55%) achieved a BCVA of 6/12 at last follow-up. Mean preoperative cylinder was 1.875 +/- 0.906 D (range, 1-3 D) and postoperatively was 1.5 +/- 1.157 D (range, 0.25-3.25 D). At last follow-up, 6 grafts were clear and 5 had failed. Mean endothelial cell count in the clear grafts at 12-month follow-up was 1078 +/- 507 cells/mm. CONCLUSIONS: DSEK provided excellent refractive and reasonable visual outcomes in our limited series, but there were frequent problems with dislocation of the donor tissue, and the graft failure rate was high. The graft failures may be linked to excessive endothelial damage, and the high dislocation rate may be linked to not filling the anterior chamber totally with air after insertion of the donor. Further development of the procedure is necessary. PMID- 17413954 TI - Agreement of physician treatment practices with the international task force guidelines for diagnosis and treatment of dry eye disease. AB - PURPOSE: To evaluate the clinical implementation of guidelines for the treatment of dysfunctional tear syndrome (DTS) developed by the International Task Force (ITF) of dry eye disease experts. METHODS: Nine physicians implemented the ITF guidelines for 3 months. Newly diagnosed patients with DTS rated their ocular symptoms and were clinically examined. Using the guidelines, physicians determined the DTS severity level (0-4, where 4 is most severe) and made their therapeutic choices. RESULTS: Of 183 patients enrolled (mean age, 65.1 years; range, 25-91 years), 67% presented without lid margin disease, and 68% had no apparent ocular surface inflammation. Symptoms were significantly more severe in patients with altered tear distribution or evident inflammation (P < 0.05). Most patients diagnosed at severity level 1 did not have lid margin disease (56/61, 92%), and inflammation was not apparent (53/58, 91%); 43% of severity level 1 patients (27/63) were treated at level 2 (therapeutic choices include unpreserved tears or topical cyclosporine). Most patients presenting with inflammation were diagnosed at severity level 2 (74%, 32/43). More than 9 in 10 severity level 2 patients were treated at level 2 (100/106; 94%). Physicians reported that 96% of their treatment recommendations were consistent with management they would have recommended if they had not consulted the ITF guidelines. They spent an average of 4.5 minutes per patient applying the guidelines. CONCLUSIONS: Implementation of the ITF guidelines was simple and not time consuming. Many practitioners chose to treat patients diagnosed at severity level 1 with treatments at level 2 that include unpreserved tears and topical cyclosporine. PMID- 17413955 TI - Delayed tear clearance in patients with conjunctivochalasis is associated with punctal occlusion. AB - PURPOSE: To evaluate the impact of punctal occlusion on ocular tear clearance and occurrence of epiphora in patients with conjunctivochalasis (CCh). METHODS: Seventy-five eyes with the diagnosis of CCh were included in the study. CCh was graded on the basis of the extent of inferior lid margin involvement as follows: 1 = one location, 2 = 2 locations, and 3 = whole lid. Patients within the 3 groups were also subdivided on the basis of the presence of punctal occlusion. The presence of epiphora, occlusion of the inferior punctum, and fluorescein clearance test (FCT) results were recorded for every patient. RESULTS: Of the 75 patients, 56 (75%) had epiphora, 47 (63%) had punctal occlusion, and 54 (72%) had delayed FCT. Twenty-two patients had grade 1 CCh, 43 had grade 2 CCh, and 10 had grade 3 CCh. Epiphora was present in 10 (45%) patients with grade 1 CCh, 38 (88%) patients with grade 2 CCh, and 8 (80%) patients with grade 3 CCh. Punctal occlusion and epiphora were observed more frequently in patients with grades 2 and 3 CCh compared with those with grade 1 CCh (Pearson chi test, P < 0.001). There was a strong correlation between delayed FCT and punctal occlusion (Spearman rank correlation analysis, r = 0.808, P < 0.001). Punctal occlusion was also correlated with the presence of epiphora (Spearman rank correlation analysis, r = 0.691, P < 0.001). CONCLUSIONS: A significant proportion of patients with CCh are symptomatic beginning with early stages of CCh. There seem to be no significant differences in any of the clinical parameters between patients with grade 2 and grade 3 CCh. Punctal occlusion seems to play an important role in the impairment of ocular tear clearance and the occurrence of epiphora. PMID- 17413956 TI - Clinical performance of a mid-viscosity artificial tear for dry eye treatment. AB - PURPOSE: We report the results of 3 studies conducted to evaluate the performance of a 1.0% carboxymethylcellulose (CMC) mid-viscosity artificial tear compared to currently marketed low-viscosity tears. METHODS: First, a single-center, double masked, randomized, crossover study was performed to compare the effect on the Ocular Protection Index (OPI) of the mid-viscosity tear compared to low-viscosity tears in 39 subjects with mild to moderate dry eye. Second, a 1-month, 2-arm, parallel, randomized double-masked clinical study assessed objective signs and subjective symptoms of dry eye in 103 subjects with mild to moderate dry eye. Third, in a 1-month home-use test, 465 artificial tear users compared the mid viscosity tear or a current low-viscosity tear to their current artificial tear. RESULTS: The OPI study showed prolonged tear breakup time and improved OPI for at least 20 minutes after instillation of the mid-viscosity tear. The low-viscosity tears showed improvements for 5 to 10 minutes. The 1-month clinical study showed a significant reduction in staining and dry eye symptoms after 1 week of treatment, with a further reduction in staining after 1 month in the mid viscosity group. Subjects provided more reports of blur with the mid-viscosity tear than with a low-viscosity tear, but equivalent overall acceptability. The home use test showed general acceptability of the mid-viscosity tear, including more subjects indicating that it was needed less frequently than their prior low viscosity tear. CONCLUSIONS: This 1% CMC mid-viscosity tear showed protection of the ocular surface after instillation and significant reduction in signs and symptoms of dry eye. Improvements were greater than with low-viscosity tears. The mid-viscosity artificial tear was rated well in comfort, duration of benefit, and general acceptability. PMID- 17413957 TI - Central corneal thickness: z-ring corneal confocal microscopy versus ultrasound pachymetry. AB - PURPOSE: To compare the repeatability and validity of corneal pachymetry by a corneal confocal microscope with a z-axis adapter (Confoscan 4.0 with z-ring adapter: z-CS4) versus ultrasound (US) pachymetry in the measurement of central corneal thickness (CCT). METHODS: CCT in 44 eyes of 44 subjects was determined with z-CS4. Z-CS4 exams were used to estimate the repeatability of thickness measurement by z-ring adapter for this confocal microscope. Intraclass Correlation Coefficient (ICC) between two different z-CS4 users was also determined. CCT in the same 44 eyes was determined with US pachymetry and measurements were compared with z-CS4 CCT. RESULTS: Z-CS4 CCT showed high intrainstrument reproducibility (ICC = 0.989; 95%CI 0.982-0.993; P < 0.0001). Mean difference among three CCT consecutive measures, in the same eye, was 0.8 +/ 11.1 microm. High correlation was found between two users (ICC = 0.896; 95%IC 0.830-0.937; P < 0.0001). Z-CS4 CCT showed high correlation with US pachymetry (ICC = 0.921; 95%CI 0.851-0.958; P < 0.0001). Mean corneal thickness determined was statistically different with the two methods (US: 512.6 +/- 65.8 microm; z CS4: 487.8 +/- 60.1 microm; P < 0.0001). CONCLUSION: Z-CS4 seems an accurate, noninvasive and reproducible technique for CCT evaluation and confirms that central cornea is thinner when measured with confocal microscopy compared to ultrasounds. PMID- 17413958 TI - Corneal thickness at high altitude. AB - PURPOSE: The eye, like other organs, is affected by the hypobaric hypoxia of high altitude. Corneal swelling is known to occur under hypoxic conditions at sea level, for instance when wearing contact lenses. The aim of this study was to measure central corneal thickness (CCT) in lowlanders ascending to altitude. METHODS: The Apex 2 medical research expedition provided the opportunity to measure CCT in 63 healthy lowlanders. The subjects arrived in La Paz, Bolivia (3700 m), where they spent 4 days acclimatizing before being driven over 2 hours to the Cosmic Physics Laboratory at Chacaltaya (5200 m), where they stayed for 7 days. CCT was measured in the early afternoon by using ultrasound pachymetry on the first, third, and seventh day at 5200 m and before and after the expedition at sea level. RESULTS: Mean CCT increased significantly from 543 microm at sea level to 561 microm on the first day at 5200 m (P < 0.001). This continued to increase to 563 microm on the third day and 571 microm on the seventh day but returned to 541 microm after descent to sea level. CONCLUSIONS: This study showed that altitude caused a significant increase in CCT in a large group of healthy lowlanders with normal corneas. This finding confirms the results of previous studies and is likely to be caused by endothelial dysfunction causing stromal swelling. This could potentially cause visual problems for high-altitude mountaineers among whom refractive surgery is popular. PMID- 17413959 TI - Topographic indications of emerging keratoconus in teenage New Zealanders. AB - PURPOSE: To screen a population of teenagers for emerging topographic signs of keratoconus (KC), with particular reference to ethnicity. METHODS: Corneal topography, visual acuity, and an environmental risk factor analysis were included in a screening program designed to identify early indications of potential KC in teenagers. Two schools from central North Island, New Zealand, were invited to participate: one with predominantly Maori/Polynesian students and the other with students of mainly European descent. RESULTS: A total of 198 Maori/Polynesian, 16.8 +/- 1.05 years of age, and 243 European students, 16.2 +/- 1.3 years of age, participated in the study. Corneal topography suggestive strongly of KC was evident in 3 Maori/Polynesian [1.2%; 95% confidence interval (CI): 0.3-4.3] and no European students (0%; 95% CI, 0%-1.5%). Anomalous corneal topography reminiscent of emerging KC in at least 1 eye were found in 19% (95% CI, 15.6%-22.9%). A significant difference (P = 0.0014) in the percentage of suspected KC was found between European students (12.9%; 95% CI, 9.2%-17.9%) and Maori/Polynesian students (26.9%; 95% CI, 21.0%-33.7%). In multivariate analyses, the significant independent predictors of probable or suspected KC included being Maori/Polynesian [odds ratio (OR) = 2.1; 95% CI, 1.25-3.54; P = 0.0052], increasing age (OR = 1.4; 95% CI, 1.10-1.80; P = 0.0067), and a history of hayfever (OR = 2.0; 95% CI, 1.16-3.59; P = 0.013). Topographical indicators that were associated significantly with suspected KC included central keratometry (P < 0.0001), astigmatism (P = 0.014), and inferior-superior asymmetry (P < 0.0001). CONCLUSIONS: Although only a longitudinal study will determine the proportion of true KC candidates, these data provide evidence of ethnicity-related differences in corneal topography in teenage New Zealanders. PMID- 17413960 TI - Corneal laceration by sharp objects in children seven years of age and younger. AB - PURPOSE: Young children are prone to develop amblyopia after penetrating injury. We sought to evaluate the management of penetrating corneal injury without intraocular foreign body (IOFB) in children 18 months. Data retrieved included all details from the initial examination, surgical procedures, amblyopia prevention measures, and final visual acuity. RESULTS: Twenty children eligible for the study were identified. The mean age was 4.3 years, and mean follow-up time was 58 months. The corneal wound size range was 1-10 mm, with 8 eyes having wounds >or=6 mm. All patients required primary surgical management. Traumatic cataract extraction was performed at the primary operation in 6 eyes, with intraocular lens (IOL) implantation in 4 eyes. Additional surgical procedures were performed in 12 eyes. Yttrium-Aluminum-Garnet (YAG) capsulotomy was performed in 3 eyes with secondary cataract. Twelve patients required spectacle or contact lens correction, and 11 patients had patching for amblyopia prevention. At the end of follow-up, visual acuity was 20/40 or better in 14 eyes, 20/50 to 20/100 in 4 eyes, no light perception (NLP) in 1 eye, and unknown in 1 eye. CONCLUSIONS: Proper management of penetrating corneal injury in young children can result in excellent visual rehabilitation. Major measures include prompt traumatic cataract extraction with either primary or secondary IOL implantation, opening of posterior capsular opacification with YAG laser, correction of refractive errors, and patching for amblyopia prevention. PMID- 17413961 TI - Corneal oxygen uptake associated with piggyback contact lens systems. AB - PURPOSE: The purpose of this study was to determine the effects of static (without blinking) and dynamic (with blinking once every 5 seconds) wear of piggyback contact lens systems on corneal oxygen uptake. METHODS: Corneal oxygen uptake rates were measured on the right eyes of 11 human subjects by using a polarographic electrode. Measurements were made for the normal open eye and after 5 minutes of wear of 4 rigid lens materials (Dk/t 0-82.5), 4 soft lens materials (Dk/t 13-122), and 16 combinations of rigid and soft lens materials. The piggyback systems were worn under both static and dynamic conditions. Repeated measures analysis of variance was used to compare oxygen uptake rates associated with the wear of the rigid lens components, soft lens components, piggyback systems, and static versus dynamic wearing conditions. Spearman correlation coefficients and regression analyses were used to examine relationships between corneal oxygen uptake rates. RESULTS: Measurable differences were found among oxygen uptake rates associated with the rigid lens components, soft lens components, and piggyback systems. Blinking resulted in no reduction in corneal oxygen uptake with the piggyback systems. Corneal oxygen uptake associated with the wear of the piggyback systems could not be predicted from those associated with the rigid and soft lens components of the systems. CONCLUSIONS: Piggyback combinations of rigid and soft lens components with the highest transmissibilities resulted in the least increase in corneal oxygen uptake beyond that of the normal open eye. PMID- 17413962 TI - Comparison of micafungin and fluconazole for experimental Candida keratitis in rabbits. AB - PURPOSE: To evaluate the efficacy of subconjunctival injection of micafungin in the treatment of experimental Candida albicans keratitis in rabbits compared with fluconazole. METHODS: In 1 eye of 24 New Zealand white rabbits, C. albicans (5 x 10 yeast cells) was inoculated in the corneal stroma. The animals were randomly assigned to 3 groups and received subconjunctival injection of 0.5 mL of 0.1% micafungin, 0.2% fluconazole, or physiologic saline once a day for 3 weeks. The eyes were examined slit-lamp biomicroscopically and histopathologically. The clinical course of fungal keratitis was compared among the 3 groups. In another 36 rabbits, a microbiological examination was performed using a quantitative isolate recovery technique, and the numbers of colony-forming units were compared among groups. RESULTS: The clinical scores were significantly lower in the micafungin group than in the other 2 groups throughout the study period (P < 0.0001 approximately P = 0.0027, Bonferroni multiple comparison). The fluconazole group showed significantly lower clinical scores than the saline group on day 18 (P = 0.0343). At the end of the study period, there were significant differences between the saline and micafungin groups (P < 0.0001), the saline and fluconazole groups (P = 0.0072), and the fluconazole and micafungin groups (P = 0.0013). Histopathologically, similar results were obtained. Moreover, the results of the microbiological examination nearly matched the clinical and histopathologic findings. CONCLUSIONS: Subconjunctival administration of micafungin was effective in the treatment of experimental Candida keratitis. Local application of micafungin to the eye would be a feasible treatment option for clinical fungal keratitis. PMID- 17413963 TI - Efficacy and safety of voriconazole as an additive in Optisol GS: a preservation medium for corneal donor tissue. AB - PURPOSE: To assess the endothelial toxicity and the microbiological efficacy of voriconazole (100 microg/mL) as an antimicrobial additive to Optisol GS. METHODS: A total of 533 donor rims were studied. One half of each donor rim was placed in standard Optisol GS and the other half rim in Optisol GS fortified with voriconazole (100 microg/mL). All rims were refrigerated for 24 hours at 3 degrees C and placed in thioglycolate broth and incubated at 37 degrees C for 7 days. A pair of donor buttons not used in transplantation was stored for 2 days in each solution and examined for endothelial changes with electron microscopy (EM). A second pair of cornea buttons was examined for toxicity by endothelial staining with 0.3% trypan blue and 0.2% alizarin red. RESULTS: Seven of 533 corneal rim cultures were positive for fungal organisms in the Optisol GS group. No rims were positive for fungal growth in the voriconazole-fortified Optisol GS medium. The difference was statistically significant (P = 0.015; Fisher exact test). There was no difference in the cellular morphology of the button stored in voriconazole fortified Optisol GS compared with Optisol GS using EM. In the bioassay, the percentage of nonviable cells in the voriconazole-fortified medium compared with the control medium was nonsignificant (P < 0.05, Student t test). CONCLUSIONS: Voriconazole seems to be safe as a fortifying agent for cornea storage medium. It significantly reduces the rate of positive fungal rim cultures and shows no signs of endothelial cytotoxicity as viewed by EM and by a bioassay of trypan blue and alizarin red. PMID- 17413964 TI - Measurement of light transmission of human limbal epithelial cells cultured on human amniotic membranes. AB - PURPOSE: To measure the light transmission properties of human limbal epithelial cell sheets (LECSs) cultured on human amniotic membranes (AMs) and compare them with those of AMs with and without amniotic epithelium. METHODS: Total light transmission of 3 kinds of tissue (LECSs, intact AMs, denuded AMs) was measured in the 250- to 800-nm range by using a spectrophotometer. RESULTS: The percent transmission of each kind of tissue decreased gradually and continually throughout the spectrum as the wavelength shortened and dropped rapidly at 300 nm to less than 20% at 250 nm. All tissues transmitted more than 70% of light in the wavelength region greater than 400 nm and more than 90% in that greater than 600 nm. The percent transmission spectrum of all tissues showed identical curves in the visible light and UV-A regions. However, the percent transmission of LECSs was lower than that of either intact or denuded AMs in the UV-B and UV-C regions. CONCLUSIONS: In the visible and UV-A light region, the percent transmission profiles of amnion-related tissues (LECSs, intact AMs, denuded AMs) are not altered by the presence of either amniotic epithelium or multilayered limbal corneal epithelium. However, the presence of multilayered limbal corneal epithelium, but not amniotic epithelium, on amniotic stroma reduced UV-B and -C transmission significantly. Further study concerning light transmission and other physical properties of LECSs is necessary to fully understand the ocular physiology of eyes grafted with such newly developed bioengineered tissues. PMID- 17413965 TI - Impact of age, systemic glucocorticoids, and progressive knee arthritis on specific mRNA levels in different areas of the rabbit cornea. AB - PURPOSE: To determine the influence of factors such as age, osteoarthritis (OA), and glucocorticoid treatment on total RNA and mRNA regulation in the cornea and how these factors differ between prepupillary and peripheral areas of the cornea. METHODS: Molecular analyses of corneal tissue were performed using rabbits of different age groups and skeletally mature animals that had undergone anterior cruciate ligament (ACL) transection, an established model of knee OA. Systemic glucocorticoids were administered to cohorts of the osteoarthritic and control animals to determine the influence of distal joint disease on the corneal response. Corneal tissue was analyzed for changes in mRNA levels for several relevant genes: collagen I, collagen III, collagen V, decorin core protein, cyclooxygenase-2 (COX-2), glucocorticoid receptor, and the housekeeping gene beta actin. RESULTS: The corneal tissue was found to have diminishing total RNA with age, which is consistent with previous studies in the literature. Interestingly, in skeletally mature animals, distal joint OA was found to affect corneal mRNA levels for several important structural and inflammatory genes (collagen I, decorin core protein, and COX-2) in a manner that progressed with OA progression. Although systemic glucocorticoid treatment did not alter mRNA levels in the normal cornea, it did counteract the changes observed early after OA induction (3 weeks) while having less of an effect in later, more established arthritis (6 weeks). CONCLUSIONS: This study reveals that distal joint OA can affect mRNA levels for several structural and inflammatory genes of the cornea, changes that seem to be suppressed by systemic glucocorticoid treatment. These findings indicate that OA has associated systemic factors that influence corneal cell metabolism. PMID- 17413966 TI - Recurrence of keratoconus in two corneal grafts after penetrating keratoplasty. AB - PURPOSE: To report the recurrence of postkeratoplasty keratoconus in 2 corneal grafts harvested from the same donor. DESIGN: Interventional case reports. METHODS: A 21-year-old-man with advanced keratoconus in his right eye and a 28 year-old-woman with corneal leucoma in her right eye underwent penetrating keratoplasty with 2 grafts coming from the same donor. Approximately 1.5 years after grafting, corneal irregularity and astigmatism caused visual acuities of the patients to decrease to counting fingers. Clinical findings and corneal topography suggested the recurrence of keratoconus. A repeat keratoplasty was performed in both patients. RESULTS: Histopathology of the excised corneal grafts was consistent with keratoconus and confirmed the preoperative diagnosis. CONCLUSIONS: Recurrence of keratoconus in a patient who had no preexisting keratoconus and in 2 corneal grafts coming from the same donor suggested transmission of the disorder from the donor instead of true recurrence. PMID- 17413967 TI - Progressive herpetic linear endotheliitis. AB - PURPOSE: To report the clinical course of a rare case of bilateral herpetic linear endotheliitis. METHODS: A 70-year-old man presented with bilateral circumferential bullous edema with stromal edema progressing centrally in the left cornea and bilateral sensorineural hearing impairment simultaneously. Serum immunoglobulin G (IgG) and IgM antibodies against herpes simplex virus type 1 (HSV1) were tested for, and aqueous humor from both eyes was examined separately using polymerase chain reaction for the presence of HSV1 DNA. RESULTS: Serum antibody titers against HSV1 were positive. In the polymerase chain reaction, the aqueous humor showed HSV1 DNA in both eyes. Forty milligrams of prednisolone was given per day and 200 mg of oral acyclovir was given 4 times daily, but corneal edema progressed. After penetrating keratoplasty surgery in the left eye, recurrent herpetic endotheliitis also seemed to occur. CONCLUSIONS: HSV-1 may cause bilateral corneal linear endotheliitis and hearing impairment simultaneously. Linear endotheliitis should be regarded as a manifestation of HSV1 corneal infection. There is a poor prognosis, and severe corneal edema can result if aggressive treatment is not used. PMID- 17413968 TI - Peripheral ulcerative keratitis-like findings associated with filgrastim. AB - PURPOSE: To describe an ocular side effect occurring with filgrastim therapy. METHODS: Retrospective case report. RESULTS: One week after starting therapy with intravenous filgrastim, a 66-year-old woman developed bilateral keratitis, evidenced by epithelial defects and peripheral corneal infiltrates. When filgrastim was discontinued, symptoms improved within 3 days and resolved within 2 weeks. CONCLUSIONS: Our patient's bilateral keratitis, which abated rapidly after discontinuation of filgrastim, may have been an adverse effect of this drug. Although the patient also took pamidronate, timing and resolution of symptoms make this a less likely culprit. Physicians should be aware of the possibility of filgrastim-associated keratitis, which can be managed by discontinuing the drug or treating with artificial tears. PMID- 17413969 TI - DNA of cytomegalovirus detected by PCR in aqueous of patient with corneal endotheliitis after penetrating keratoplasty. AB - PURPOSE: Corneal endotheliitis often leads to severe endothelial dysfunction and can be caused by herpes simplex virus (HSV), varicella zoster virus (VZV), and other viruses (eg, the mumps virus). We report a case of corneal endotheliitis caused by cytomegalovirus (CMV) that developed after a penetrating keratoplasty. METHODS: A complete ophthalmologic examination was performed on a patient with corneal endotheliitis that developed after a penetrating keratoplasty. To determine the cause of the endotheliitis, polymerase chain reaction (PCR) was used to amplify the DNA of HSV, VZV, and CMV in samples of the aqueous humor. RESULTS: Slit-lamp biomicroscopy showed a moderate stromal edema in the upper temporal part of the transplanted cornea along with keratic precipitates (KPs) arranged in a coin-shaped pattern. Repeated treatments with steroids and acyclovir were only temporarily successful. PCR detected the DNA of CMV in an aqueous sample, and the treatment was switched to topical and systemic application of ganciclovir. This resulted in the disappearance of the KPs and resolution of the stromal edema within 2 weeks. CONCLUSIONS: From the PCR results and the favorable response to ganciclovir, the corneal endotheliitis was most likely caused by cytomegalovirus in this case. PMID- 17413970 TI - Three-dimensional optical coherence tomography of granular corneal dystrophy. AB - PURPOSE: To describe 3-dimensional optical coherence tomography (OCT) findings in a case of granular corneal dystrophy. METHODS: A 1310-nm high-speed swept-source OCT prototype was used to image the 3-dimensional structures of the cornea of a patient with granular corneal dystrophy. RESULTS: Three-dimensional OCT clearly showed multiple highly reflective lesions, corresponding to the deposits in the cornea. Volume of the deposits within the central 1.5 mm of the cornea was 0.11 mm and occupied 2.98% of the corneal volume. CONCLUSIONS: Three-dimensional OCT allows objective and noninvasive assessment of granular corneal dystrophy. PMID- 17413971 TI - Acute bilateral diffuse corneal opacity in a child. AB - PURPOSE: To report a case of acute bilateral corneal opacity related to bacterial blepharoconjunctivitis. METHODS: Interventional case report. An 18-month-old boy had bilateral red eyes with discharge and periorbital eczema. Bilateral corneal opacity suddenly developed 1 week later. RESULTS: All clinical assessments and laboratory evaluations of viral infection and autoimmune profiles were negative except for growth of coagulase-negative staphylococcus and propionibacterium acnes from conjunctival discharge. After systemic and topical antibiotic treatment, the opacity and inflammation of both eyes decreased. The left cornea became clear, but the right eye had a sequela of corneal opacity with stromal neovascularization and conjunctival scarring. Penetrating keratoplasty was performed in the right eye 6 months later. Histology at that time showed neovascularization in the deep stroma with perivascular mononuclear cell infiltration and decreased endothelial cells. CONCLUSIONS: Acute bilateral corneal opacity is rare but may occur in children with bacterial blepharokeratoconjunctivitis. Ocular surface inflammation may be induced by bacterial toxin reaction. Combined use of antibiotics and steroids may save the vision in such patients. PMID- 17413972 TI - Bilateral prominent schwalbe ring in the anterior chamber in a patient with axenfeld-rieger syndrome and megalocornea. AB - PURPOSE: To report the unusual presentation of bilateral Schwalbe rings suspended in the anterior chambers of a patient with Axenfeld-Rieger syndrome. METHODS: A 37-year-old man with bilateral decreasing visual acuity underwent slit-lamp examination, dark room gonioscopy, and photographic documentation. RESULTS: Prominent bilateral Schwalbe rings and peripheral iridocorneal strands were observed in both anterior chambers. No systemic abnormalities were found. CONCLUSIONS: The position of the Schwalbe ring in patients with Axenfeld-Rieger syndrome can vary in appearance from almost normal to displaced markedly. It remains to be seen whether these variations reflect genetic differences. PMID- 17413974 TI - Contact lens-induced infiltrates. PMID- 17413975 TI - Benign endometrial hyperplasia sequence and endometrial intraepithelial neoplasia. AB - Endometrial "hyperplasia," as currently diagnosed, includes the changes caused by an abnormal hormonal state and those caused by a separate category of monoclonal premalignant disease. The appearance of the disease in these 2 functional categories is discontinuous, permitting more specific diagnosis of the condition using the terms "benign endometrial hyperplasia" and "endometrial intraepithelial neoplasia" (EIN), respectively. Benign endometrial hyperplasia involves the entire endometrial compartment and, with protracted estrogen exposure, shows the progressive development of cysts, remodeled glands, vascular thrombi, and stromal microinfarcts. They are best construed as a sequence of changes whereby the appearance at any single time point is uniquely dependent on the preceding combination and the duration of hormonal exposures. In contrast, the premalignant clone of an EIN lesion is characteristically offset from the background endometrium by its altered cytology and crowded architecture. The use of an internal standard for cytology assessment, combined with the distinctive topography of a clonal process, enables the diagnosis of EIN lesions with a long term cancer risk 45-fold greater than that of their benign endometrial hyperplasia counterparts. The resolution of hormonal and premalignant subsets of traditional "endometrial hyperplasias" is possible using redefined diagnostic criteria, enabling patient therapy to be appropriately matched with the underlying disease mechanisms. PMID- 17413976 TI - Difficulties in assessing the depth of myometrial invasion in endometrial carcinoma. AB - The depth of myometrial invasion (DMI) is one of the most important prognostic indicators and determinants of therapy in endometrial cancer. There are well documented problems in recognizing DMI. We examined 100 previously diagnosed endometrioid endometrial carcinomas in hysterectomy specimens, reassessed DMI, and explored morphological features that complicated appraisal of myometrial invasion. The DMI was different from the original measurement in 29% of cases. Twelve percent of all cases (40% of cases with measurement discrepancies) involved differences in the assignment of invasion categories (noninvasive, < or =50% myometrial invasion, and >50% myometrial invasion). Nearly all endometrial cancers originally diagnosed as invasive were considered noninvasive on review. We examined whether the distribution of stromal metaplasia, noninvasive patterns (exophytic tumors, irregular endomyometrial junctions, and adenomyosis), and myometrial invasion patterns were different in cases with and without measurement discrepancies. Irregular endomyometrial junctions, exophytic tumors, and adenomyosis tended to coexist and were more common in the cases with DMI discrepancies. Although there seemed to be a relationship between smooth muscle metaplasia and exophytic tumors, it did not appear that smooth muscle metaplasia was significantly more common in cases with measurement difficulties. However, cases with extensive smooth muscle metaplasia posed problems with assessment of myometrial invasion. Patterns of myometrial invasion other than the conventional destructive pattern were sufficiently uncommon as to not impact on DMI measurement in large numbers of cases. Measuring the DMI is usually uncomplicated, but additional scrutiny should be paid to cases involving exophytic tumors, irregular endomyometrial junctions, adenomyosis, and extensive stromal smooth muscle metaplasia. PMID- 17413977 TI - Expression of estrogen and progesterone receptors in smooth muscle metaplasia of rectovaginal endometriosis. AB - To investigate expression of estrogen receptors (ER) and progesterone receptors (PR) in smooth muscle metaplasia (SMM) outside the endometriotic foci of rectovaginal endometriosis (RVE). One hundred and ninety-five specimens were obtained from the rectovaginal areas of the 63 patients who were underwent laparoscopic surgery for RVE. The patients were divided into 3 groups: a gonadotropin-releasing hormone (GnRH) agonist group, a non-GnRH group, including a proliferative phase group, and a secretory phase group. Expression of ER and PR in the rectovaginal tissues of RVE were determined using immunohistochemical methods. Smooth muscle metaplasia occurred in 172 specimens (88.2%), and ER and PR expression were found in the smooth muscle cells in the SMM areas outside the endometriotic foci of RVE. The expression of ER and PR in the GnRH agonist group were significantly lower than those in the non-GnRH agonist group. This is the first report demonstrating ER and PR in the smooth muscle cells in SMM outside the endometriotic foci of RVE. The ER and PR were expressed in the SMM areas, but these receptors were not recognized in fibrotic areas. We could identify the expression ratio of these receptors during each menstrual phase, with or without administered GnRH agonist. PMID- 17413978 TI - Plasma cell myeloma presenting as abnormal uterine bleeding. AB - We report a case of a 53-year-old woman who presented with anemia and abnormal uterine bleeding. Endometrial and cervical biopsies revealed infiltration by large atypical cells, which were characterized as neoplastic plasma cells. Further investigations revealed systemic plasma cell dyscrasia (i.e., plasma cell myeloma). We therefore report a case of plasma cell myeloma, initially presenting as abnormal uterine bleeding. PMID- 17413979 TI - Carcinomas of ovary and lung with clear cell features: can immunohistochemistry help in differential diagnosis? AB - Metastatic lung carcinomas with clear cell morphology can be confused with primary ovarian clear cell carcinomas. We performed immunohistochemical stains in 14 cases of non-small cell lung carcinomas with clear cell features and 14 cases of ovarian clear cell carcinomas using a panel of markers, including thyroid transcription factor 1 (TTF-1), carcinoembryonic antigen (CEA), Wilms tumor gene 1, octamer-binding transcription factor 4 (OCT-4), cancer antigen 125 (CA-125), estrogen receptor, and progesterone receptor. Among non-small cell lung carcinomas with clear cell features, 87.5% of adenocarcinomas (or 50% overall frequency in lung carcinomas) were positive for TTF-1, whereas none of the ovarian clear cell carcinomas were positive (P = 0.002). All 14 ovarian clear cell carcinomas stained for CA-125 as compared with 1 non-small cell lung carcinoma (P < 0.001). On the other hand, 85% of non-small cell lung carcinomas stained for CEA, whereas none of the ovarian clear cell carcinomas did (P < 0.001). Interestingly, 4 ovarian clear cell carcinomas (28%) showed positive staining for the germ cell marker OCT-4. Either lung or ovarian carcinomas stained for Wilms tumor gene 1, estrogen receptor, or progesterone receptor very infrequently; and the difference between the 2 groups was not statistically significant. Our results suggest that an immunohistochemical panel consisting of TTF-1, CEA, CA-125, and OCT-4 is helpful in distinguishing most pulmonary and ovarian carcinomas with clear cell features. PMID- 17413980 TI - Expression of extracellular matrix proteins in ovarian serous tumors. AB - The aims of this study were to perform a comprehensive expression analysis of the genes encoding extracellular matrix proteins and to investigate the expression pattern in one of these proteins, syndecan 1, in normal ovarian epithelium as well as benign and malignant ovarian serous tumors. Gene expression of 16 different extracellular matrix proteins was analyzed in ovarian serous tumors based on serial analysis of gene expression database. Semiquantitative reverse transcription-polymerase chain reaction was used to validate the serial analysis of gene expression result from each gene. As compared with normal ovarian surface epithelium, we found overexpression of syndecan 1, collagen type IV alpha 2, elastin microfibril interfase located protein 1, and laminin 5 in ovarian serous carcinomas. Syndecan 1 was selected for further study as it has not been well characterized in ovarian cancer and the syndecan 1 antibody was available for immunohistochemistry. Using a syndecan 1-specific monoclonal antibody, we demonstrated that syndecan 1 was expressed in 30.4% of high-grade serous carcinomas, 29.7% of low-grade carcinomas and serous borderline tumors, but none of benign serous cystadenomas and ovarian surface epithelium. Although both high grade and low-grade serous carcinomas had a similar percentage of syndecan 1 positive cases, the immunointensity in high-grade carcinoma was significantly higher than that in low-grade carcinomas and serous borderline tumors (P = 0.007). In summary, ovarian carcinomas exhibit up-regulated expression of several extracellular matrix proteins, and syndecan 1 represents a novel tumor-associated marker in ovarian serous carcinomas. PMID- 17413981 TI - Poly(adenosine diphosphate-ribose) polymerase expression in serous ovarian carcinoma: correlation with p53, MIB-1, and outcome. AB - This study investigated the expression of poly(adenosine diphosphate-ribose) polymerase (PARP) in a cohort of ovarian serous carcinomas by immunohistochemistry with regard to outcome, clinicopathologic parameters, proliferation as assessed by MIB-1 labeling indices (LIs), and p53 immunoexpression. Formalin-fixed, paraffin-embedded archival tissues of 50 ovarian serous carcinomas were immunostained with antibodies to PARP, MIB-1, and p53. In addition, 10 benign serous cystadenomas and 10 typical serous borderline ovarian tumors were included in the PARP immunostudy. Immunostaining for PARP was scored with regard to quantity and intensity of positively stained nuclei as negative, low, or strong. The MIB-1 LIs were quantitated as the percentage of positively stained nuclei in 1000 nuclei. For p53, at least 10% of tumor cells had to display nuclear staining. The expression of PARP was scored negative in all serous cystadenomas and low in serous borderline ovarian tumors. Strong PARP expression was determined in 38 cases (76%), and low expression in 12 cases (12%) of ovarian serous carcinomas; MIB-1 staining was noted in all cases (mean, 44.2; range, 10.8-66.5), positivity for p53 in 39 cases (78%). The PARP immunoreactivity increased with the International Federation of Gynecology and Obstetrics stage (P = 0.0075), as well as p53 positivity (P = 0.0141) and MIB-1 LIs (P = 0.0102), with grade determined after Malpica et al. (P = 0.0445) but not with grade assessed after Shimizu et al. (P = 0.1495). A trend for poor outcome was observed in patients whose tumors displayed high levels of PARP immunoexpression (P = 0.0196, log-rank test). This study indicates that PARP expression is frequently upregulated in ovarian serous carcinomas, related with MIB-1 LIs and p53 expression, and may serve as a marker of aggressive behavior with prognostic value. PMID- 17413982 TI - Clinical and pathological predictive factors in women with adult-type granulosa cell tumor of the ovary. AB - Granulosa cell tumor (GCT) is a rare neoplasm hallmarked by a very indolent course and late recurrences. Although numerous clinical and pathological parameters have been implicated as prognostic factors for GCT, their role remains controversial. We performed a retrospective study at our institution where we identified 48 patients with GCT from our tumor registry. Demographic and clinical course information was recorded from the medical record. Twenty of 48 formalin fixed, paraffin-embedded blocks were retrieved from archived specimens. Pathological parameters such as nuclear atypia, mitotic count, Ki-67 index using immunohistochemistry, and quantitative DNA ploidy were performed. DNA aneuploidy by quantitative method was associated with patients' overall survival. The degree of nuclear atypia, mitotic count, Ki-67 index, and DNA aneuploidy was not predictive of tumor recurrence. Multi-institutional collaboration is imperative to create a comprehensive national database for investigation into ways that may better indicate prognosis in these patients. PMID- 17413983 TI - Mixed ovarian germ cell tumor in a BRCA2 mutation carrier. AB - BRCA2 germ-line mutations confer an increased risk of developing breast and ovarian cancer. We report the occurrence of a mixed ovarian germ cell tumor (GCT) (50% embryonal carcinoma, 20%-25% choriocarcinoma, 10%-15% dysgerminoma, and 10% 15% immature teratoma) in a 33-year-old Ashkenazi Jewish woman, carrier of the BRCA2:6174delT mutation. The mutation is also present in the patient's father, who was diagnosed with breast cancer at age 59 and with prostate cancer at age 69. This is the first report of a GCT in a BRCA2 mutation carrier; there was one previous report of an ovarian dysgerminoma in a BRCA1 carrier. Molecular analysis of the proband's tumor DNA revealed there was no loss of heterozygosity of the wild-type allele in the tumor, as is usually the case for epithelial BRCA-related ovarian tumors. This suggests either that biallelic inactivation of BRCA2 is not required for GCT development or that this is a chance event unrelated to the presence of the mutation. PMID- 17413984 TI - Primary ovarian adenofibromatous neoplasms with mucin-containing signet-ring cells: a report of 2 cases. AB - The presence of mucin-containing signet-ring cells in an ovarian neoplasm is considered strong evidence for a metastatic rather than primary tumor. The clinicopathologic features of 2 ovarian adenofibromatous neoplasms with mucin containing signet-ring cells (AFN-SRs), proven by the long-term follow-up of 10 and 15 years to be primary tumors, are presented as exceptions to this general guideline. Both of these tumors were initially strongly favored to be metastatic adenocarcinoma by expert consultants in gynecologic pathology, which underscores the diagnostic difficulty that these rare and unusual neoplasms can pose. Although long-term follow-up of AFN-SRs is still necessary to exclude the possibility of metastases, the following constellation of features can correctly suggest a primary neoplasm rather than a Krukenberg-like metastatic process at the time of presentation: (i) the appearance of fibromatous stroma as an integral part, as opposed to reactive part, of the neoplasm, (ii) a semiorganized rather than completely haphazard pattern to the placement of the epithelial elements, with a circumscribed noninfiltrative interface with ovarian parenchyma, (iii) the absence of angiolymphatic invasion, (iv) tumor unilaterality, and (v) the lack of a known primary source. The virtual absence of literature on these low-grade malignancies, coupled with the occurrence of 2 such tumors within a 7-year period at a single community hospital, suggests that many AFN-SRs continue to be misdiagnosed as metastatic adenocarcinoma. PMID- 17413985 TI - Low-grade fibromyxoid sarcoma presenting clinically as a primary ovarian neoplasm: a case report. AB - Low-grade fibromyxoid sarcoma is an uncommon, deceptively bland mesenchymal neoplasm that typically occurs in the deep soft tissues of the proximal extremities of young to middle-aged patients. Intra-abdominal low-grade fibromyxoid sarcomas are distinctly rare. We describe the first reported example of this sarcoma involving the ovary. The 42-year-old patient presented with progressing abdominal pain and urinary frequency. Computed tomographic imaging of the abdomen and pelvis showed a 14-cm left-sided pelvic mass. The patient underwent surgical resection and, intraoperatively, a left ovarian mass was identified that extended to both the left hypogastric artery and the left ureter. The resected specimen consisted of a 17-cm tan, fleshy mass containing grossly recognizable nodules of gelatinous, myxoid tissue and a small rim of normal ovary. Microscopic examination of the neoplasm revealed a cytologically bland spindle cell proliferation set in a collagenized stroma that abruptly transitioned to nodules of a myxoid stroma with a well-formed capillary vasculature, features characteristic of low-grade fibromyxoid sarcoma. On the basis of the diagnosis, the patient was clinically staged as a retroperitoneal sarcoma with secondary ovarian involvement. The patient has stable residual disease 11 months postoperatively. This case adds to the literature of intra abdominal low-grade fibromyxoid sarcoma and expands the list of malignant mesenchymal neoplasms that may involve the ovary. PMID- 17413986 TI - Immunohistochemical and functional biomarkers of value in female genital tract lesions: a systematic review with statistical meta-analysis. PMID- 17413987 TI - Clonality analysis and human papillomavirus infection in squamous metaplasia and atypical immature metaplasia of uterine cervix: is atypical immature metaplasia a precursor to cervical intraepithelial neoplasia 3? AB - Atypical immature squamous metaplasia (ISM) of the uterine cervix often has histological features that overlap with the histological characteristics of high grade cervical intraepithelial neoplasia. To identify the cellular basis and clinical significance of atypical immature metaplasia (AIM), 10 cases of AIM were analyzed for the clonal status, and the presence of human papillomavirus (HPV) infection. The physical status of HPV was also evaluated in HPV type 16 (HPV-16) positive cases. Squamous metaplasias with no nuclear atypia (29 mature squamous metaplasias [SMs]) and a single case of ISM were analyzed as a control. Nine AIMs, 20 SMs, and a single case of ISM were informative for clonal analysis. Monoclonal composition of the lesion was demonstrated in 8 (89%) of 9 AIMs, but only in 2 (10%) of 21 cases of SM without atypia (AIM vs SM + ISM, 8/9 vs 2/21; P < 0.0001). High-risk HPV was detected in 6 (60%) of 10 AIMs, all were HPV-16, but only in 3 (13%) of 24 SMs with no atypia (2/23 SM and 1/1 ISM). The frequency of high-risk HPV infection was also significant between AIMs and SM with no atypia (6/10 vs 3/24; P < 0.001). Among the cases, which were informative for clonal analysis, all 5 AIMs positive for high-risk HPV were monoclonal in composition. Physical status of HPV was examined in HPV-16-positive cases. Human papillomavirus type 16 was present as a mixture of episomal form and integrated form in 4 of 6 AIMs. These observations imply that unlike SMs with no atypia, which arises as a result of reactive or inflammatory process, lesions with the histological characteristics of AIM may be indeed true precursors of cervical carcinoma. PMID- 17413988 TI - Identification of potential prognostic markers for vulvar cancer using immunohistochemical staining of tissue microarrays. AB - The aim of this study is to determine immunohistochemical markers with prognostic significance for disease-specific survival in patients with squamous cell cancer of the vulva. The study material consisted of slides and paraffin blocks of 50 vulvectomy specimens. A tissue microarray was constructed and stained with 16 antibodies. The impact of lymph node metastases, size of tumor, vascular space involvement, and the marker expression on disease-specific survival was calculated. In univariate analysis lymph node metastases, tumor size more than 4 cm, vascular space involvement, strong cyclooxygenase 2 expression, and absent Caspase 3 expression were significantly associated with disease-specific survival. In a multivariate analysis, poor disease-specific survival is independently associated with absent Caspase 3 expression (hazard ratio, 0.2; 95% confidence interval, 0.04-0.97; P = 0.045). Five-year survival was 86% in patients with tumors positive for Caspase 3 (n = 20) and drops to 64% in patients with Caspase 3-negative tumors (n = 30). In this test set, cyclooxygenase 2 and Caspase 3 seem to be immunohistochemical markers with prognostic significance for vulva cancer. The results have to be validated. PMID- 17413989 TI - Lymphoma-like lesion of the uterine cervix: report of 12 cases of a rare entity. AB - Lymphoma-like lesion of the female genital tract is rare. We report 12 cases of lymphoma-like lesions of the cervix in patients ranging from 27-54 years of age (mean 41). The commonest clinical presentation was post-coital bleeding (8), which was followed by vaginal bleeding (2) and leukorrhagia (4). Grossly, the lesions were either polypoid (8) or ulcerated (4). On histological examination, the lesions were eroded and involved the superficial mucosa 2-12 mm (mean 4 mm) in depth. They comprised sheets of dense populations of predominantly large lymphoid cells admixed with small lymphocytes, plasma cells, and neutrophils. Follicle formation was occasionally seen. Immunostaining revealed the majority of the large cells were B cells (CD20(+), CD79a(+)) with no aberrant CD5 and CD43 expression. The lymphoid cells in the follicle were CD10(+) and bcl6(+) but negative for bcl-2. Cyclin D1 was negative. There was no immunoglobulin light chain restriction and polymerase chain reaction for T cell receptor-gamma chain gene and immunoglobulin heavy chain gene demonstrated polyclonal patterns. In situ hybridization for EBER and high risk HPV 6/11 and 16/18 were negative. All patients were well with one case developing local recurrence in the follow-up period up to 7 years. PMID- 17413990 TI - Villoglandular adenocarcinoma of the cervix: two new cases with morphological and molecular study. AB - In this paper, we report 2 new cases of villoglandular papillary adenocarcinoma (VGPA) of the cervix, a rare, well-differentiated form of cervical adenocarcinoma. Both patients were without medical complications or history of oral contraceptive use and were nonsmokers. Extended hysterectomy was performed in both cases. Morphological criteria for a correct pathological diagnosis were emphasized. Immunohistochemical analysis was performed to clarify the phenotype of the neoplasms. Moreover, for the first time, we probed to establish if VGPA could be correlated to human papilloma virus (HPV) and herpes virus (HSV) types 1 and 2, using polymerase chain reaction amplification of tumoral DNA. Both neoplasms showed positivity for B72.3, Ca-125, carcinoembryonic antigen, keratin 7, and p16(INK4a) protein. Vimentin, P53, estrogen, and progesterone receptors, instead, were negative. Molecular study by polymerase chain reaction amplification of tumor DNA revealed a strong positive signal for HPV-DNA and no signal for HSV-DNA. It is reasonable to conclude that our cases of VGPA, in accordance with other examples reported in literature, are due to HPV infection. Behavioral cofactors, such as HSV infection (types 1 and 2), oral contraceptive use and smoking, involved in the pathogenesis of other cervical malignancies, can be excluded for the present cases. PMID- 17413991 TI - Chronic coinfection with hepatitis B and hepatitis C viruses in an Italian population of HIV-infected patients. PMID- 17413993 TI - Neuroanesthesiology review--2006. AB - In this article, we will provide a review of the 2006 literature of interest to those readers who provide perioperative care to patients with neurologic disease. This evaluation of the literature is not intended to be comprehensive, nor were systematic criteria used to include or exclude articles. Instead, the authors attempted to highlight those articles of greatest clinical relevance or those that provided unique insights into the physiology, pharmacology, and pathomechanisms of neurologic function for practicing clinicians and clinician investigators. This article focuses on intracranial hemorrhage, anesthetic considerations in neurosurgical patients, cerebral hemodynamics, electrophysiologic monitoring, neuroprotection, and traumatic brain injury. PMID- 17413994 TI - Equi-MAC concentrations of halothane and isoflurane do not produce similar bispectral index values. AB - Minimum alveolar concentration (MAC) has been traditionally used to measure the potency of an inhalational anesthetic agent. Recently, bispectral index (BIS) derived from the frontal cortical electroencephalogram has been used frequently for quantifying the hypnotic component of anesthesia. The present study was designed to examine the BIS values produced by equi-MAC concentrations of halothane and isoflurane. In 34 patients undergoing spinal surgery, BIS and spectral edge frequency (SEF95) were recorded at 3 different concentrations of halothane and isoflurane--namely 0.5, 0.75, and 1.0 MAC. The measurements were made both during wash-in and wash-out phases of the anesthetic agent. Eighteen patients received halothane and 16 received isoflurane. Heart rate, mean arterial pressure, oxygen saturation, and end tidal carbon dioxide pressure values were not different between the 2 groups at various MAC concentrations of the anesthetic agents. BIS and SEF95 values decreased significantly with increasing concentrations of both the anesthetic agents (P<0.001). At any given MAC concentration of the anesthetic, BIS and SEF(95) values were significantly lower under isoflurane compared with halothane anesthesia both during wash-in and wash out phases (P<0.001). For a given anesthetic agent, BIS values were comparable at equi-MAC concentrations during wash-in and wash-out phases. In conclusion, BIS values are significantly lower under isoflurane compared with halothane anesthesia at similar MAC concentrations. For a given anesthetic agent and a given MAC concentration, the BIS values are similar during wash-in and wash-out phases of anesthesia. PMID- 17413995 TI - The application of near-infrared oximetry to cerebral monitoring during aneurysm embolization: a comparison with intraprocedural angiography. AB - Near-infrared spectroscopy (NIRS) has been used to monitor regional cerebral oxygen saturation (rSO2) in patients at risk of cerebral desaturation during surgical and neurointerventional procedures. However, the quantitative capabilities of the method have been questioned, as has its validation compared with jugular bulb oximetry. Here, we compare NIRS data acquired during coil embolization procedures with incidence of vasospasm as detected from angiography. Thirty-two subarachnoid hemorrhage patients underwent embolization. Bilateral SomaSensor strips (Invos 4100, Somanetics) were affixed to the forehead at constant anatomic positions, avoiding frontal sinuses and scalp hair. Mean arterial pressure, SaO2, end-tidal pCO2, temperature and Hb were held within a narrow range during the procedure. Ipsilateral angiography was performed every 10 to 15 minutes. An independent neuroradiologist classified any vasospasm in the parent vessel as mild (25% baseline), moderate (50%), severe (75%), or total (100%). Of all, 15/32 (46.9%) patients developed spasm; in 2 it was severe or total. There was no significant association between World Federation of Neurological Surgeons grade and baseline rSO2 signal (either ipsilateral or contralateral to the side of the aneurysm) (P=0.598). There was no significant association between side of aneurysm and baseline rSO2 signal (P=0.243). However, episodes of angiographic spasm were strongly associated with reduction in trend ipsilateral NIRS signal (P<0.001); furthermore, the degree of spasm (especially more than 75% vessel diameter reduction) was associated with a greater reduction in same-side NIRS signal (P<0.001) (2-level random effects regression model, Stata 8.2, Stata Corp, TX). NIRS may have a useful role to play in the detection of cerebral desaturation secondary to vasospasm during neuroendovascular procedures. PMID- 17413996 TI - Midline-shift corresponds to the amount of brain edema early after hemispheric stroke--an MRI study in rats. AB - Vasogenic brain edema formation is a serious complication in hemispheric stroke. Its space-occupying effect can lead to midline-shift (MLS), cerebral herniation, and death. Clinical studies indicate that quantification of MLS can predict cerebral herniation and subsequent death at early time-points, even before clinical deterioration becomes apparent. The present experimental study was designed to determine the relation between MLS, absolute edema volume, lesion size, and clinical findings in a rat stroke model. Middle cerebral artery occlusion was performed in 24 rats using the suture technique. Clinical evaluation and magnetic resonance imaging (MRI) (Bruker PharmaScan 7.0T) was performed 24 hours later. Lesion volume, the volume-increase within the affected hemisphere (%HEV), and MLS were quantified on T2-weighted images. The absolute increase of hemispheric water content (DeltaH2O) was determined in a subgroup using the wet-dry method (n=12). MLS correlated significantly with the total amount of brain edema (magnetic resonance imaging study: r=0.82; P<0.01; wet-dry analysis r=0.80; P<0.01). MLS correlated only moderately with T2-lesion volume (r=0.55; P<0.01). No significant correlation could be detected between MLS and clinical scores (r=0.26; P>0.05). MLS thus quantitatively reflects the amount of vasogenic brain edema within the affected hemisphere at early time-points. MLS quantification can be regarded as an easily assessable and valid global quantitative parameter for brain edema and thus might facilitate the surgical and nonsurgical management of edema in acute stroke patients. PMID- 17413997 TI - Intra-arterial drug delivery: a concise review. AB - The therapeutic potential of intra-arterial (IA) drug delivery to the brain has received limited attention in the last decade. In the 1980s, efforts to treat brain tumors with IA chemotherapy, the leading application of this technology, yielded modest results. Poor control of tissue drug concentrations and the potential risk of permanent neurologic injury further prevented the wider use of IA drugs. Yet, IA drugs were anecdotally used for treating a wide spectrum of brain diseases. Recent advances in endovascular technology and the increased safety of angiographic procedures now compel us to reevaluate IA drug delivery. This review describes the pharmacologic principles, applications, and pitfalls of IA drug delivery to the brain. PMID- 17413998 TI - Anesthetic management for surgical placement of greater occipital nerve stimulators in the treatment of primary headache disorders. AB - We will present a case series looking at the anesthetic management of 4 patients undergoing surgical placement of greater occipital nerve stimulators for chronic headache. The anesthetic technique described uses total intravenous anesthesia with the guidance of Bispectral Index Scale to smoothen the transition between different anesthetic depths required for this novel operation. PMID- 17413999 TI - Anesthetic considerations of selective intra-arterial nicardipine injection for intracranial vasospasm: a case series. AB - Cerebral vasospasm after subarachnoid hemorrhage can decrease cerebral blood flow with the potential for stroke. Induction of Triple-H therapy (hypertension, hypervolemia, and hemodilution) is an accepted medical therapy to decrease the delayed cerebral ischemia related to vasospasm. Recently selective intra-arterial injection of nicardipine during angiography has also been proposed as a therapeutic modality for the management of distal vasospasm not amenable to balloon angioplasty. We are reporting the hemodynamic changes in 11 patients who underwent this procedure. A retrospective chart review of 15 procedures in 11 patients showed a significant change in blood pressure after the injection of nicardipine. Blood pressure changes were not different between sexes, but increase in heart rate was higher for females. A significantly higher drop in systolic blood pressure but not for diastolic blood pressure or mean arterial pressure after the injection was seen in patients who were not intubated in the intensive care unit before the procedure. Selective intra-arterial injection of nicardipine during angiography can cause significant hemodynamic instability and requires supportive management by the anesthesiologist. PMID- 17414000 TI - Effectiveness of an intravascular cooling method compared with a conventional cooling technique in neurologic patients. AB - Fever is common among neurologic patients and is usually treated by antipyretic drugs and external cooling. An alternative method for temperature management may be an intravascular approach. The aim of the study was to compare the effectiveness and the therapeutic costs of this new method with conventional treatment in neurologic patients. Twenty-six patients who suffered from subarachnoid hemorrhage or traumatic brain injury with febrile episodes were included the study and were randomized into 2 different groups. In the "Conventional" group, fever was treated with antipyretic drugs and/or surface cooling techniques to achieve a body core temperature of 36.5 degrees C. In the "CoolGard" group, patients were treated with an intravascular cooling catheter (Coolgard, Alsius, CA). We compared the effectiveness of these 2 approaches by calculating the mean deviation from 36.5 degrees C during a 48-hour period (fever burden). We found a significant difference in the fever burden [CoolGard: -0.49 to 1.22 (median -0.06) degrees C vs. Conventional: 1.05-2.34 (median 1.41) degrees C, P<0.05]. Costs varied significantly between the CoolGard and the Conventional groups, with markedly higher daily costs in the CoolGard group [CoolGard: 15 to 140 US dollars (USD) (median 39 USD) vs. Conventional: 1 to 9 USD (median 5 USD), P<0.05]. The effectiveness of the intravascular cooling catheter is excellent compared with conventional cooling therapies. PMID- 17414003 TI - Neostigmine-induced increase in EEG activity following severe traumatic brain injury. PMID- 17414004 TI - Spectral entropy monitoring in a patient undergoing awake craniotomy. PMID- 17414005 TI - Guided growth for angular correction: a preliminary series using a tension band plate. AB - The classic treatment of pathological angular deformities of the extremities is corrective osteotomy; however, osteotomies require hospitalization, pain management, immobilization, and delayed weight bearing. The associated risks, inconvenience, and cost of osteotomy make hemiepiphysiodesis or guided growth an attractive option. Although stapling has a long and proven track record, reported drawbacks related to implant failure, including migration or breakage of staples, have led some to abandon this technique. This report describes a prospective series of 34 consecutive patients who presented with a total of 65 deformities (femur and/or tibia) due to a variety of pathological conditions and who underwent guided growth using a nonlocking extraperiosteal 2-hole plate and screws. This technique relies upon the tension band principle rather than physeal compression. With follow-up ranging from 14 to 26 months (from implantation) in this series, 32 of 34 patients (63 deformity levels) have corrected to neutral at a mean of 11 months and the hardware has been removed. The observed rate of correction was approximately 30% more rapid than noted with stapling, and there have been no permanent growth arrests. Four patients with bilateral idiopathic genu valgum experienced rebound deformity and have since undergone repeat guided growth using the same technique. Only 2 patients with adolescent Blount disease have experienced insufficient correction; each may need a corrective osteotomy of the tibia. Having prevented 63 (97%) of 65 osteotomies in this series of patients, it is evident that guided growth holds promise for postponing if not preventing more invasive surgery.These patients will be observed up to maturity to support my conclusion that the concept of osteotomy as a first resort and criterion standard has become outdated. PMID- 17414006 TI - Multiplanar deformity analysis of untreated Blount disease. AB - Although varus malalignment of the proximal tibia is the primary pathology in Blount disease, other deformities may exist. To assess multiplanar lower limb deformities, children with previously untreated early- and late-onset Blount disease who subsequently needed surgical correction were identified. Preoperative frontal and sagittal plane deformity analysis using Paley's methodology and rotational profile assessment using prone clinical examination were performed by a single examiner. Results were compared between the 2 groups and with uninvolved limbs within each group. Additionally, rotational profile of the lower limb was compared with age-matched values. Over an 8-year period, 60 limbs (40 patients) including 26 with early-onset and 34 with late-onset Blount met the inclusion criteria. Although both groups exhibited proximal tibial varus, procurvatum, and internal torsion, patients with early-onset Blount disease had greater severity. Unlike the younger patients, approximately one third of the varus malalignment of the affected extremity was attributed to the distal femur in the late-onset patients. Neither group showed any significant deformity of the proximal femur and distal tibia or sagittal plane deformity of the distal femur. There was a correlation between the severity of varus malalignment of the limb with magnitude of proximal tibial deformities in both groups and with distal femoral varus in the late-onset group of patients. Multiplanar deformity analysis is a valuable tool in the comprehensive evaluation of children with Blount disease. PMID- 17414007 TI - Tibial aplasia-hypoplasia and ectrodactyly in monozygotic twins with a discordant phenotype. AB - Tibial hemimelia with ectrodactyly is reported for the first time in monozygotic female twins with a positive family history for limb anomalies. This very rare autosomal dominant condition is known to be associated with a highly variable phenotype, as illustrated by the distinct clinical presentations, despite identical genotype. PMID- 17414008 TI - Professional opinion concerning the effectiveness of bracing relative to observation in adolescent idiopathic scoliosis. AB - OBJECTIVE: To determine if community equipoise exists concerning the effectiveness of bracing in adolescent idiopathic scoliosis. BACKGROUND DATA: Bracing is the standard of care for adolescent idiopathic scoliosis despite the lack of strong reasearch evidence concerning its effectiveness. Thus, some researchers support the idea of a randomized trial, whereas others think that randomization in the face of a standard of care would be unethical. METHODS: A random of Scoliosis Research Society and Pediatric Orthopaedic Society of North America members were asked to consider 12 clinical profiles and to give their opinion concerning the radiographic outcomes after observation and bracing. RESULTS: An expert panel was created from the respondents. They expressed a wide array of opinions concerning the percentage of patients within each scenario who would benefit from bracing. Agreement was noted concerning the risk due to bracing for post-menarchal patients only. CONCLUSIONS: : This study found a high degree of variability in opinion among clinicians concerning the effectiveness of bracing, suggesting that a randomized trial of bracing would be ethical. PMID- 17414009 TI - Early experience with fluorine-18 sodium fluoride bone PET in young patients with back pain. AB - PURPOSE: Skeletal positron emission tomography (PET) with fluorine-18 (18F) sodium fluoride (18F NaF) is an alternative to technetium-99m (99mTc)methylene diphosphonate (MDP) scintigraphy. Experience with pediatric PET is sparse, primarily in oncology. This study assesses the role of 18F NaF in evaluating young patients with back pain. METHODS: Ninety-four 18F NaF PET scans were performed in 94 patients (27 males, 67 females; mean age, 15 years; range, 4-26 years) with back pain. Three-dimensional PET acquisition was performed 30 minutes after administration of 18F NaF (2.1 MBq/kg; maximum, 148 MBq). Radiation doses are presented for 18F NaF and 99mTc MDP. RESULTS: 18F NaF PET revealed a possible cause of back pain in 55% (52/94). Fifteen patients had 2 or more potential sources of back pain. Diagnoses by PET were pars interarticularis/pedicle stress (34%), spinous process injury (16%), vertebral body ring apophyseal injury (14%), stress at a transitional vertebra-sacral articulation (7%), and sacroiliac joint inflammation/stress (3%). Comparing 18F NaF PET with 99mTc MDP scintigraphy, time between injection and scanning was shorter (0.5 hours vs 3 hours), radiation dosimetry was similar (3.5 mGy vs 2.8 mGy effective dose for a 55-kg patient for 18F NaF and 99mTc MDP, respectively), and cost of radiopharmaceutical was higher. CONCLUSIONS: 18F NaF bone PET can detect a variety of skeletal abnormalities in young patients with back pain. Relative to 99mTc MDP, images are of higher resolution. Total time from tracer administration to completion is shorter, and radiation dosimetry is similar. Higher cost for 18F NaF may be offset by enhanced patient throughput. PMID- 17414010 TI - Oxygen cost versus a 1-minute walk test in a population of children with bilateral spastic cerebral palsy. AB - Difficulties with direct measurement of oxygen (O2) consumption have led to the use of simpler proxy measures to estimate energy efficiency in children with cerebral palsy. This study aimed to investigate the relationship between a fast 1 minute walk test and O2 cost in children with bilateral spastic cerebral palsy. Eighty-four children (31 girls, 53 boys; mean age, 10 years 11 months; Gross Motor Function Classification System level I n = 8, level II n = 41, level III n = 21, level IV n = 14) were eligible to participate in the study. Seventy-seven children completed the walk test and 47 children completed the O2 consumption protocol. Results showed significant differences between Gross Motor Function Classification System levels for both tests (P < 0.001) and a significant moderate quadratic relationship between the distance covered during the 1-minute fast-walk test and the net O2 cost (adjusted r = 0.477; P < 0.001). The 1-minute fast-walk test may provide useful information on the efficiency of gait in children with bilateral spastic cerebral palsy in the absence of more sophisticated equipment; however, further work is required to address its reliability and responsiveness. PMID- 17414011 TI - Use of a dynamic foot pressure index to monitor the effects of treatment for equinus gait in children with cerebral palsy. AB - The purpose of this study is to introduce and describe a newly developed index using foot pressure analysis to quantify the degree of equinus gait in children with cerebral palsy before and after injection with botulinum toxin. Data were captured preinjection and 12 weeks postinjection. Ten children aged 2(1/2) to 6(1/2) years took part (5 boys and 5 girls). Three of them had a diagnosis of spastic diplegia and 7 of congenital hemiplegia. In total, 13 limbs were analyzed. After orientation and segmentation of raw pedobarographic data, we determined a dynamic foot pressure index graded 0 to 100 that quantified the relative degree of heel and forefoot contact during stance. These data were correlated (Pearson correlation) with clinical measurements of dorsiflexion at the ankle (on a slow and fast stretch) and video observation (using the Observational Gait Scale). Pedobarograph data were strongly correlated with both the Observational Gait Scale scores (R = 0.79, P < 0.005) and clinical measurements of dorsiflexion on a fast stretch, which is reflective of spasticity (R = 0.70, P < 0.005). We demonstrated the index's sensitivity in detecting changes in spasticity and good correlation with video observations seems to indicate this technique's potential validity. When manipulated and segmented appropriately, and with the development of a simple ordinal index, we found that foot pressure data provided a useful tool in tracking changes in patients with spastic equinus. PMID- 17414012 TI - Advanced skeletal maturity in ambulatory cerebral palsy patients. AB - INTRODUCTION: Both advanced and delayed bone age relative to chronological age have been described in non ambulatory children with moderate to severe cerebral palsy (CP). The purpose of our study was to assess skeletal maturation in an ambulatory CP population and determine the affects of body mass index (BMI), type of CP, and Gross Motor Function Classification System (GMFCS) on skeletal maturity. METHODS: A retrospective chart and radiograph review was performed on 51 patients with ambulatory CP. A control group of 50 patients was also analyzed. Age, sex, height, weight, type of CP, and GMFCS were recorded from the chart. The height and weight were used to calculate BMI. Bone age was determined using the Oxford method. Statistical analysis for the data included descriptive statistics with bivariate and multivariate regression analyses. Significance was determined as P < 0.05. RESULTS: There were 26 boys and 25 girls. All CP patients were independent ambulators. The mean chronological age was 7.1 years for boys and 8.6 years for girls. The mean bone age was 9.9 years for boys and 10.6 years for girls. Overall, 48 (94%) of 51 patients had advanced bone age compared with chronological age. Bone age was significantly advanced compared with chronological age for boys (P = 0.033) and showed a trend toward significance in girls (P = 0.079). Bone age was advanced compared with our control population in both sexes. In multivariate analysis, quadriplegic CP type showed a trend toward significance (P = 0.066), and GMFCS III was significantly associated with advanced bone age in boys (P = 0.011). In girls, quadriplegic CP type and BMI of less than 15 were significantly associated with advanced bone age (P < 0.05 in both). CONCLUSION: Our results demonstrated that most of the ambulatory CP patients had advanced bone age compared with chronological age. Quadriplegic CP type in boys and girls contributed to advanced bone age. GMFCS III and a low BMI also contributed to advanced bone age in boys and girls, respectively. Understanding factors that lead to either delayed or advanced skeletal maturation is important in planning the appropriate timing for orthopaedic surgical intervention. PMID- 17414013 TI - Femur fracture in preschool children: experience with flexible intramedullary nailing in 72 children. AB - Spica cast immobilization is the preferred treatment for closed femur fracture in preschool children. Flexible intramedullary nailing (FIN) is an alternative treatment but has never been specifically evaluated in this age group. A retrospective analysis of 72 children (mean age, 4.1 years; age range, 1.5-5.9 years) with 73 femur fractures treated by means of FIN was performed. The mean length of stay in hospital was 9.1 days (range, 1-46); weight bearing was started at a mean of 16.4 days (range, 1-60 days) after surgery. Follow-up was available in 62 children (mean, 36.7 months; range, 4-124 months). Complications included early distal nail exteriorization in 9 children (12.3%) during the first 10 years of the study. More than 1 cm of femur overgrowth was noted in 6 children (8.2%). Preschool children with femur fracture treated by means of FIN benefited from short hospital stay, early mobilization and weight bearing. Technique-related complications could be avoided. Long-term follow-up is mandatory in these children. PMID- 17414014 TI - Osteochondral flap fracture of the olecranon in a child. AB - Osteochondral flap fracture in a child is rare. A 7 year-old boy with an osteochondral flap fracture is reported. Intraoperatively, the osteochondral fragment in the olecranon fossa was connected to a part of the medial collateral ligament. The authors suppose that this type of fracture occur not only as a result of a shearing force of the trochlea but also by a traction force through the medial collateral ligament when a complete tear of the medial collateral ligament was avoided for any reason. Furthermore, the immaturity of bone of the child may result in avulsion fracture, not in injury of the distal part of the ligament. Osteosynthesis with poly-L-lactic acid pins was useful to reacquire good elbow function. PMID- 17414015 TI - Cyst index: a nonpredictor of simple bone cyst fracture. AB - PURPOSE: Treatment is indicated for simple bone cysts (SBC) with high risk of fracture. The cyst index was described as a method to determine if a cyst is at high risk or low risk of fracture. STUDY DESIGN: The cyst index of 32 femoral and humeral SBCs was determined by 8 reviewers. The risk group cyst index was compared with whether a fracture took place. Sensitivities, specificities, and positive and negative predictive values were calculated across varying cutoff levels. Intraobserver and interobserver reliability testing for 10 cysts was made. RESULTS: Receiver operator curves demonstrate that no cutoff value gave an acceptable level of both sensitivity and specificity. The mean value for the cyst index was significantly different for different observers (P < 0.05). CONCLUSIONS: This study does not validate the cyst index to be an accurate predictor of fracture. The test has low intraobserver and interobserver reliability. CLINICAL RELEVANCE: The cyst index cannot reliably discriminate between the patients that will fracture and those that will not. PMID- 17414016 TI - Near-infrared spectroscopy identifies compartment syndrome in an infant. AB - Acute compartment syndrome is a surgical emergency requiring immediate intervention to limit tissue and nerve injury. Although the measurement of compartment pressure is frequently performed, there is controversy regarding the pressure at which fasciotomy should be performed. Near-infrared spectroscopy (NIRS) uses the same technology as pulse oximetry to estimate tissue oxygenation. To date, NIRS is used most commonly to estimate cerebral oxygenation during intraoperative anesthetic care. The authors present a 1-month-old infant who developed an acute compartment syndrome of the right lower extremity after cardiac surgery. In addition to the measurement of compartment pressures, the diagnosis of compartment syndrome was confirmed by NIRS with a value of 15% in the involved leg versus 40% to 50% in the noninvolved contralateral lower extremity. The potential use of this modality in identifying compartment syndrome is reviewed. PMID- 17414017 TI - Biomechanical stability of bioabsorbable screws for fixation of acetabular osteotomies. AB - The purpose of this study was to compare the biomechanical stability of triple innominate osteotomies fixed with either bioabsorbable or stainless steel screws. Triple innominate osteotomies were performed on composite hemipelves and fixed with either three 4.5-mm bioabsorbable screws or three stainless steel 4.5-mm screws. Two screws were placed from the iliac wing into the acetabular fragment, and 1 screw was placed from below the acetabular fragment into the iliac wing. Eight specimens for each screw type were biomechanically tested in an anatomical position (replicating weight bearing) and in a flexed and abducted position (replicating spica cast positioning). Specimens were cyclically loaded between 10 and 450 N to simulate the hip contact force in this population. Lower screws were then removed, and specimens were tested under identical conditions. Fragment displacement (mm) and construct stiffness (N/mm) were compared with a 2-way analysis of variance (P < 0.05). There were no significant differences between materials for fragment displacement or construct stiffness. Anatomical position showed significantly less displacement than spica position for both materials. Initial displacement in the spica position was significantly less during lower loads for stainless steel fixation. Bioabsorbable screws demonstrate comparable biomechanical stability to stainless steel screws in anatomical and spica positions at physiological loads. Flexion and abduction of the femur adversely affect the stability of the construct for both materials. Bioabsorbable screws behave similarly to steel screws when stabilizing triple innominate osteotomies and would have the advantage of not requiring a second surgery for screw removal. Confirmation of biocompatibility should be completed before widespread clinical application. PMID- 17414018 TI - Screw head impingement after in situ fixation in moderate and severe slipped capital femoral epiphysis. AB - In situ stabilization remains the standard of care in the treatment of stable slipped capital femoral epiphysis (SCFE). Screw placement perpendicular to the physis has shown satisfactory results with minimal complications. A prominent screw head may produce femoral acetabular impingement and pain after in situ fixation in severe SCFE. We performed a biomechanical study to establish whether screw head impingement occurs after in situ fixation of SCFE and to define the anatomy of slip severity and screw head position that may lead to impingement. A femoral neck dome osteotomy was created in a human cadaveric model simulating 2 conditions: a moderate and severe SCFEs. We tested the specimens after in situ fixation perpendicular to the simulated physis. The simulated SCFEs and normal control were tested through a full arc of motion. Coverage of the femoral head by the labrum was evaluated at 90 degrees of flexion using fluoroscopy. Impingement occurred at 70 degrees of hip flexion in the simulated moderate SCFE, and at 50 degrees of flexion in the severe simulated SCFE. Anteroposterior fluoroscopy revealed that screw heads lateral to the intertrochanteric line were unlikely to impinge on the acetabulum. Screw head impingement occurred with in situ fixation perpendicular to the physis in simulated moderate and severe SCFEs. Anteroposterior radiographs appear helpful in identifying a hip at risk for screw head impingement after in situ fixation. Alternative in situ fixation techniques (screw head resting lateral to the intertrochanteric line on the anteroposterior radiograph) may decrease the rate of screw head impingement in moderate and severe SCFEs. PMID- 17414019 TI - Unique case of polydactyly and a new classification system. AB - Polydactyly of the hands or feet is a common birth deformity. We recently encountered a female infant with a case of a crossed type 1 polydactyly with a mixed polydactyly of the feet. A mixed and crossed polydactyly is a rare finding with only one other reported case. This is the first report of crossed and mixed polydactyly of the feet presenting with 7 complete toes on each foot without syndactyly. In addition to a discussion of the treatment, this case has lead us to propose a more complete and less confusing classification system. PMID- 17414020 TI - Wound complications from idiopathic clubfoot surgery: a comparison of the modified Turco and the Cincinnati treatment methods. AB - PURPOSE: Treatment protocols using the Turco and the Cincinnati incisions are widely used for the surgical correction of clubfoot deformity. However, it is unclear which surgical approach leads to fewer wound problems. We therefore sought to determine which treatment method led to a lower incidence of wound complications. STUDY DESIGN: A retrospective chart review of 217 consecutive patients (308 feet) who underwent a primary posteromedial release for the treatment of idiopathic clubfoot under the age of 24 months via either the modified Turco or Cincinnati treatment methods was used to document the incidence of postoperative wound complications. The modified Turco protocol involved immediate postoperative casting in neutral, whereas the Cincinnati method involved staged casting with the foot initially in equinus, then to neutral with a cast change 7 days later. RESULTS: A significantly lower incidence of wound complications was seen in the Cincinnati treatment group when compared with the modified Turco treatment method (6.9% vs 19.6%, respectively, P < 0.003). When patients were stratified based on immediate versus staged postoperative casting methods, there was a significantly lower incidence of wound complications (P < 0.05) in feet in the Cincinnati treatment group versus the modified Turco treatment method; however, the statistical populations were markedly unequal. Among all feet treated with the Cincinnati method, patients who underwent a staged cast change had significantly fewer wound problems when compared with those who underwent immediate casting with the foot in neutral (5.1% vs 16.7%, respectively, P < 0.04). CONCLUSIONS: In the surgical correction of idiopathic clubfoot, the incidence of wound complications is significantly decreased with the use of the Cincinnati treatment method rather than the modified Turco treatment protocol. Whether this effect is a result of the incision or the postoperative casting protocol is unclear. PMID- 17414021 TI - Abnormal scarring with keloid formation after osteochondroma excision in children with multiple hereditary exostoses. AB - INTRODUCTION: Multiple hereditary exostoses (MHE) is an autosomal dominant condition characterized by numerous cartilage-capped exostoses/osteochondromas in areas of actively growing bone. Abnormal scarring with keloid formation after osteochondroma excision in children with MHE has not been previously described. METHODS: A retrospective double-cohort study was undertaken to determine if children with MHE have a higher rate of abnormal scarring with keloid formation after osteochondroma excision when compared with those with solitary osteochondroma. In the initial phase, all consecutive children with MHE that underwent excision of osteochondroma with a minimum 2-year postoperative follow up were identified. A control group of age-matched cases of solitary osteochondroma was subsequently identified. All patients were interviewed for wound healing problems and noncosmetic scarring. All patients with unsatisfactory scars were asked to send pictures and/or were invited for follow-up. Data were statistically analyzed. RESULTS: Eighty-three surgeries were performed in 25 patients with MHE, whereas 25 surgeries were performed in 25 patients with solitary osteochondroma. Twelve keloid scars were noted in 7 patients with MHE, and no keloids were noted in any of the patients in the solitary group. Diagnosis of MHE was a statistically significant risk factor for formation of keloids after surgery (P < 0.05). DISCUSSION: Abnormal scarring with keloid formation after osteochondroma excision in MHE has not been previously reported. Although this study has limited numbers, the results demonstrate a statistically significant correlation between keloid formation and MHE. The risk for abnormal scarring and keloid formation should be discussed with all patients before surgery. PMID- 17414022 TI - Angular measurement error due to different measuring devices. AB - BACKGROUND: Angular measurements are commonly used in orthopaedic surgery. No study has addressed measurement variability due to the measurement device itself. It was the purpose of this study to assess measurement variability of articulated versus fixed devices. METHODS: Three articulated and 4 fixed goniometers were randomly selected. Thirty-two different angles, ranging from a few degrees to nearly 180 degrees, were drawn with a standard soft-lead marking pencil. The angles were measured by 5 different observers with 7 different goniometers on 2 separate occasions separated by a minimum of 3 weeks. We wished to determine whether the variability of any goniometer was within a +/-1-degree range. RESULTS: There were 1023 (91.3%) absolute differences of 1 degree or less and 97 (8.7%) of more than 1 degree. Intraobserver agreement was 92.0%; 96.9% for fixed and 84.0% for hinged goniometers. Interobserver agreement was of 90.7%; 96.2% for the fixed and 83.4% for the hinged goniometers. Intragoniometer agreement was 91.3%; 96.9% for fixed and 84.0% for hinged goniometers. Intergoniometer agreement was 87.0% with fixed goniometers demonstrating better agreement than hinged goniometers. The overall intraobserver and interobserver measurement variability was +/-2.33 and +/-2.26 degrees, respectively; the overall intragoniometer and intergoniometer measurement variability was +/-2.26 and +/ 2.30 degrees, respectively. For fixed goniometers, the intergoniometer measurement variability is +/-2.0 degrees, for hinged goniometers, +/-2.9 degrees, and when using both fixed and hinged goniometers, +/-2.4 degrees. Thus, the fixed goniometers are the ideal type with overall better agreement and measurement variability. CONCLUSION: Any one orthopaedic surgeon should use the same goniometer at all times, preferably a fixed type, so that measurement variability can be reduced by +/-2.0 degrees. If a physician uses a particular published measurement variability from the literature in which different goniometers were used, then the intraobserver measurement variability will be less than the published value by approximately +/-2 degrees. This is important when faced with the question of a change in an angular measurement being a true change or simply a reflection of measurement error. PMID- 17414023 TI - Use of plain radiography to optimize skeletal outcomes in children with type 1 Gaucher disease in Brazil. AB - BACKGROUND: Gaucher disease is the most common lysosomal storage disease and is caused by deficient production and activity of the lysosomal enzyme beta glucosidase (glucocerebrosidase), resulting in progressive accumulation of glucosylceramide (glucocerebroside) in lysosomes of cells of the reticuloendothelial system in the spleen, liver, and marrow. Clinical manifestations include anemia, thrombocytopenia, hepatosplenomegaly, and bone complications, including bone pain, bone marrow infiltration, lytic lesions, osteopenia, pathological fractures, and avascular necrosis. Early, adequate, and sustained treatment with enzyme replacement therapy (ERT) available since 1991 can change the natural history of the disease, particularly in children. Skeletal complications are usually the major source of disease morbidity and disability and although magnetic resonance imaging and dual-energy x-ray absorptiometry densitometry are recommended for monitoring, these are not readily available in all countries. METHODS: We describe 18 Brazilian children with type 1 Gaucher disease with bone involvement who were followed with plain radiography for at least 8 months after beginning imiglucerase ERT (initial dose, 15-60 U/kg body weight/15 days). Bone involvement noted by plain radiograph included marrow infiltration, osteopenia, pathological fractures, osteonecrosis, lytic lesions, and Erlenmeyer flask deformity. Patients were questioned about bone crises. RESULTS: Patients were followed for up to 10 years (mean follow-up, 4 years and 4 months +/- 3 years and 3 months). Bone changes were visible by plain radiographs in all patients. Clinical and radiological improvement was noted in 13 (72%) of 18 patients; bone lesions worsened in 5 (28%) of 18 patients. The final ERT dose for the 13 patients who improved was 55 +/- 10 U/kg (range, 30-60 U/kg), and the final ERT dose for the 5 who worsened was 29 +/- 2 U/kg (range, 26-30 U/kg); this difference was statistically significant (P < 0.03). CONCLUSIONS: When other imaging technologies are not available, skeletal response to ERT in children with type 1 Gaucher disease can be monitored effectively by plain radiography. Higher doses of ERT (50-60 U/kg /15 days) may be required for improvement of skeletal manifestations. CLINICAL EVIDENCE: Case series; level 4. PMID- 17414024 TI - Central physeal arrests as a manifestation of hypervitaminosis A. AB - Vitamin A is necessary for synthesis of visual pigments and required in appropriate amounts for membrane stability. Acute hypervitamin A intoxication can lead to increased intracranial pressure, vomiting, and lethargy. Chronic excessive intake of vitamin A can lead to pruritus, muscle and bone tenderness, and failure to thrive. Reported effects of hypervitamin A intoxication on bone include osteoporosis, fracture, cortical thickening, and metaphyseal irregularity. We are reporting on a case of central physeal arrest in the distal femur, proximal tibia, and distal tibia after excessive intake of vitamin A. PMID- 17414025 TI - Growing rod techniques in early-onset scoliosis. AB - BACKGROUND: The surgical treatment of severe early-onset scoliosis (EOS) is controversial. Obtaining and maintaining deformity correction, achieving adequate spinal growth, allowing lung development, and the high complication rate make surgical treatment very challenging. Growing rods are the most common method of management. METHODS: Currently, there are 3 systems being used for the surgical treatment of EOS: single growing rod, dual growing rods, and the vertical expandable titanium prosthetic rib implant. Each system has its advantages and disadvantages. These are presented and discussed in this review. RESULTS: The current clinical and radiographic results indicate that all 3 techniques can be effective in the surgical management of EOS. Vertical expandable prosthetic titanium rib (VEPTR), which is not considered a true growing rod system, is particularly effective in congenital scoliosis with fused ribs. CONCLUSIONS: The current expandable spinal implant systems appear effective in controlling progressive EOS, allowing for spinal growth and improving lung development. All have a moderate complication rate, especially rod breakage and hook displacement. PMID- 17414027 TI - Body mass index in patients with slipped capital femoral epiphysis. PMID- 17414028 TI - Percutaneous reaming of simple bone cysts in children followed by injection of demineralized bone matrix and autologous bone marrow. PMID- 17414030 TI - Management of unstable/acute slipped capital femoral epiphysis. PMID- 17414031 TI - Lateral column lengthening as treatment for planovalgus foot deformity in ambulatory children with spastic cerebral palsy. PMID- 17414033 TI - Maintenance of certification: an update. PMID- 17414034 TI - Otosclerosis of the incus. AB - OBJECTIVE: To report a case of a patient with otosclerosis of the incus. PATIENTS: A 61-year-old woman with a progressive hearing loss on her left ear and a computed tomographic scan of the temporal bone revealing an expansible lesion of the incus. INTERVENTIONS: The ossicle was removed by using a transtympanomastoid approach; the ossicular chain was reconstructed using a titanium partial ossicular replacement prosthesis. MAIN OUTCOME MEASURE: The diagnosis of the disease was obtained by means of histopathologic examination of the specimen. RESULTS: The patient obtained a good postoperative hearing result. The histopathologic examination of the specimen documented an otosclerosis of the incus. CONCLUSION: Otosclerotic involvement of the middle ear ossicles, apart from footplate, was very rarely mentioned. Most subjects were incidentally diagnosed postmortem by means of examination of specimens from temporal bone collections. The diagnosis and treatment of a patient with otosclerosis of the incus is exceptional; however, otosclerosis should be considered in the differential diagnosis of expansible lesions of the ossicles. PMID- 17414035 TI - Intratympanic methylprednisolone for sudden sensorineural hearing loss. AB - OBJECTIVE: Corticosteroids are commonly used for the treatment of sudden sensorineural hearing loss (SSHL). In this study, the effectiveness of intratympanic (IT) corticosteroid injection was studied and compared with a control group on patients with SSHL who failed systemic corticosteroid treatment. MATERIALS AND METHODS: A total of 19 patients as a retreatment group (RG) and 18 patients as a control group (CG), all failed high-dose intravenous and oral corticosteroid treatments, were included in this study. These patients were invited back, and IT methylprednisolone was injected five times via 3-day intervals in RG and followed-up for a mean period of 24.9 months (range, 7-30 mo). Audiological evaluations were performed initially, a week after the completion of the injections, monthly in the following first 3 months, and at the end of follow-up period in RG. The CG was followed-up for 3 months after the completion of systemic corticosteroid treatment without any additional drug administration. RESULTS: The mean age was 52.6 years (range, 20-79 yr) in RG and 59.9 years in CG. The mean pure-tone average for speech frequencies (500-4,000 Hz) at baseline audiogram and at the first month, at the third month, and at last controls were 65.2 (range, 43-102 dB), 45.4 (range, 23-77 dB), 43.6 (range, 30-77 dB), and 44.5 (range, 33-77 dB) dB, respectively, in RG. The mean pure-tone averages for speech frequencies (500-4,000 Hz) at the end of systemic treatment and at third-month control were 63.5 (range, 44-98 dB) and 59.0 (range, 40-100 dB) dB, respectively, in CG. The hearing gain that is equal to or more than 10 dB was achieved in 14 patients (73.6%) at the last control in RG. No hearing gain could be detected in the CG. No serious side effect was observed with IT treatment. CONCLUSION: We conclude that IT methylprednisolone injection provides more significant hearing improvement for patients that failed with previous high dose systemic corticosteroid administration than systemic corticosteroid treatment alone. So it may be the first-step medical treatment of idiopathic SSHL alone or at least may be combined with the systemic corticosteroid administration. PMID- 17414036 TI - Computed tomography and/or magnetic resonance imaging before pediatric cochlear implantation? Developing an investigative strategy. AB - OBJECTIVE: To investigate and compare the usefulness of preoperative magnetic resonance (MR) imaging and high-resolution temporal bone computed tomography (HRCT) in pediatric cochlear implant candidates. STUDY DESIGN: Prospective. SETTING: Tertiary referral center. PATIENTS: A cohort of 92 pediatric patients with profound hearing. Inclusion criteria were MR, computed tomography, and cochlear implantation. INTERVENTION. DIAGNOSTIC: All patients had preoperative imaging of the petrous temporal bone (HRCT, T2-weighted fast spin echo, axial 3D Fast Imaging Employing Steady-state Acquisition [FIESTA] MR) and brain (Fast Fluid-attenuated Inversion-recovery [FLAIR] MR). MAIN OUTCOME MEASURE(S): Overall prevalence of inner ear dysplasias in this population and comparison of detection rates between HRCT, T2 Fast Spin Echo (FSE), and FIESTA MR sequences. RESULTS: Radiological abnormalities were observed in 32 and 59% of MR and HRCT temporal bone, respectively. Synchronous intracranial findings were noted in 40% on brain MR. Common vestibulocochlear nerve was observed in 3% ears and directed side of implantation. Consistent discrepancies noted on HRCT were inability to diagnose early obliterative labyrinthitis and presence of the cochlear nerve in the internal auditory canal. With respect to MR, enlarged vestibular aqueducts and narrow cochlear nerve canals were consistently under identified. CONCLUSION: Dual modality imaging with HRCT and MR of petrous bone and MR brain in the precochlear implant pediatric population detects abnormalities related to deafness, which would not otherwise be found using either modality alone. There is overlap between the imaging modalities in the type of abnormalities detected, and we present a case for selective use of HRCT within a diagnostic algorithm, using the patient risk factors we have identified. PMID- 17414037 TI - Percutaneous cochlear access using bone-mounted, customized drill guides: demonstration of concept in vitro. AB - HYPOTHESIS: Percutaneous cochlear access can be performed using bone-mounted drill guides that are custom made on the basis of preintervention computed tomographic scans. BACKGROUND: We have previously demonstrated the ability to use image guidance based on fiducial markers to obtain percutaneous cochlear access in vitro. A simpler approach that has far less room for application error is to constrict the path of the drill to pass in a predetermined trajectory using a drill guide. METHODS: Cadaveric temporal bone specimens (n = 8) were affixed with three bone-implanted fiducial markers. The temporal bone computed tomographic scans were obtained and used in planning a straight trajectory from the mastoid surface to the cochlea without violating the boundaries of the facial recess, namely, the chorda tympani, the incus buttress, and the facial nerve. These surgical plans were used to manufacture a customized drill guide by means of rapid prototyping (MicroTargeting Platform; FHC Inc.; Bowdoinham, ME, U.S.A.) that mounts onto anchor pins previously used to mount fiducial markers. The specimens then underwent traditional mastoidectomy with facial recess. The drill guide was mounted, and a 1-mm drill bit was passed through the guide across the mastoid and the facial recess. The course of the drill bit and its relationship to the boundaries of the facial recess were photographed and measured. RESULTS: Eight cadaveric specimens were subjected to the study protocol. In seven of eight specimens, the drill bit trajectory was accurate; it passed from the lateral cortex to the lateral wall of the cochlea without compromise of any critical structures. In one specimen, the access to the middle ear was achieved, but the incus was hit by the drill. The average shortest distance +/- standard deviation from the edge of the drill bit to the boundaries of the facial recess was 0.78 +/ 0.56 mm (chorda tympani), 2.00 +/- 1.06 mm (incus buttress), and 1.27 +/- 0.54 mm (facial nerve). CONCLUSION: Our study demonstrates the ability to obtain percutaneous cochlear access in vitro using customized drill guides manufactured on the basis of preintervention radiographic studies. PMID- 17414038 TI - Indications for and outcomes of mastoid obliteration in cochlear implantation. AB - OBJECTIVE: To review the indications, efficacy, and long-term outcomes of mastoid obliteration in cochlear implant surgery. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Seventeen patients who underwent a mastoid obliteration procedure to facilitate the insertion of a cochlear implant between 1978 and 2005. INTERVENTION: Mastoid obliteration procedure before cochlear implantation. MAIN OUTCOME MEASURES: Revision rate of the mastoid obliteration and cochlear implantation, postoperative audiometric scores (consonant-nucleus-consonant words/phonemes, Central Institute for the Deaf sentences, City University New York sentences in quiet and in noise), and incidence of complications. RESULTS: There were 17 patients with a median age of 60 years (range, 3-79 yr). Eight patients required mastoid obliteration for active chronic suppurative otitis media before cochlear implantation. Another 8 patients had existing mastoid cavities requiring obliteration (modified radical [n = 5] and fenestration cavities [n = 3]). A single patient with a sclerotic mastoid and an anterior sigmoid sinus underwent obliteration because of inadequate surgical access. The technique of obliteration was radical mastoidectomy with eustachian tube occlusion, blind sac closure of the external auditory canal, and cavity obliteration with either temporalis muscle flap (n = 15) or abdominal fat (n = 2). Cochlear implantation and mastoid obliteration were performed as a two-stage procedure in 10 patients and as a single-stage procedure in 7. Two patients required revision of the mastoid obliteration. At follow-up, all patients had stable obliterated cavities. Fifteen patients obtained significant improvement in speech discrimination scores, whereas 2 patients obtained some benefit from the cochlear implant through the perception of environmental sounds. CONCLUSION: For patients with chronic suppurative otitis media or existing mastoid cavities, the obliteration with temporalis muscle or abdominal fat is an effective technique to facilitate safe cochlear implantation. PMID- 17414039 TI - Impact of Jacobson's (tympanic) nerve sectioning on middle ear functions. AB - OBJECTIVE: The purpose of this study was to evaluate the influence of sectioning of the Jacobson's (tympanic) nerve on middle ear functions. METHOD: Twenty-five adult New Zealand rabbits were included in this study. The Jacobson's nerve was cut in the left ear of the rabbits (study group), whereas only a small mucosal incision was performed while keeping the Jacobson's nerve intact in their right ear (control group). After the operation, the ears were assessed both otomicroscopically and histopathologically on Days 30, 60, and 90. RESULTS: On otomicroscopy, retraction pockets were observed in 48 and 4% of the ears in the study and control groups, respectively (p < 0.001). Middle ear effusion was observed in 56 and 12%, respectively (p < 0.01). Histopathologically, an inflammation in the middle ear mucosa was present in all ears in the study group, whereas it was present only in 20% of the control ears (p < 0.001). Goblet cells were observed in 48 and 20% in the study and control groups, respectively (p < 0.04). In the study group, the otomicroscopic and histopathologic findings were more prominent on Day 60 compared to Day 90 (p < 0.05). CONCLUSION: Tympanic glomus cells seem to act as middle ear chemosensory organs and are involved in the regulation of middle ear aeration. Disruption of these neural elements such as Jacobson's nerve negatively impacts on middle ear functions and may result in atelectasis. PMID- 17414040 TI - Tympanic membrane perforation among adults in West Africa. AB - CONTEXT: The knowledge of variations of the tympanic membrane (TM) perforations with the climatic changes in the West African subregion would help clinicians in its prevention and management. OBJECTIVE: To analyze the pattern of clinical presentations and associated features of TM perforation in adults in West Africa. DESIGN: A prospective study. SETTING: Tertiary referral centre, University hospital. PATIENTS OR OTHER PARTICIPANTS: Thirty-five (35) consecutive adults with TM perforations during a 1-year period had clinical evaluation of each TM using head mirror, video otoscopy, and micro-otoscopy. MAIN OUTCOME MEASURES: Clinical presentations and associated features of TM perforations. RESULTS: Thirty-five patients, 20 (57%) men and 15 (43%) women, with 42 perforated TMs were examined. Twenty-eight (80%) patients had unilateral perforations. Infection was responsible for 90.5% of cases, and trauma was responsible for the rest. Locations of perforations were central (29; 69.1%), anteroinferior (4; 9.5%), posteroinferior (4; 9.5%), anterosuperior (3; 7.1%), and posterosuperior (2; 4.8%). The sizes of the perforations ranged from 1.2 to 83.2%. Large sizes of 25% and more were found to occur in humid and wet seasons, and also, clinical presentations of otorrhea (65.6%), otalgia (51.5%), tinnitus (37.1%), and ear itching (34.4%) seemed to worsen. CONCLUSION: Most TM perforations result from infection and are preventable via appropriate health education. Posterosuperior perforation is rare, and this is probably one of the factors making choleasteatoma uncommon in West Africa. Approximately 83.3% of TM perforations measure more than 25% in size and appear during the wet humid season of the year. These findings are important for both local and foreign otolaryngologists who may be practicing in this subregion of the world. PMID- 17414041 TI - A longitudinal study on pediatric myringoplasty. AB - OBJECTIVES: Observational study on pediatric myringoplasty to examine the success rate in young and older children. PATIENTS AND INTERVENTION: Fifty-one first-time primary myringoplasties were analyzed on children aged 4 to 13 years. MAIN OUTCOME MEASURES: Using life-table analysis, the survival course of the graft in young children (aged 4-8 yr) was compared with those in older children (aged 9-13 yr). The effect of age on various outcome parameters, including otitis media with effusion/atelectasis, discharge, and hearing, was analyzed using the 12-month data. RESULTS: The 3-year graft take rate was 83.8% (95% confidence interval: 70.2, 97.5%). At 12 months, only 63.0% (42.4, 80.6%) were free of otitis media with effusion/atelectasis and discharge and preserved their hearing. There is no evidence of a difference in outcome in young and older children. CONCLUSION: There was no evidence that age influenced surgical outcome in pediatric myringoplasty. Although the graft take rate was more than 80%, true success was only found in two thirds of cases. PMID- 17414042 TI - Semicircular canal function before and after surgery for superior canal dehiscence. AB - OBJECTIVE: To characterize semicircular canal function before and after surgery for superior semicircular canal dehiscence (SCD) syndrome. STUDY DESIGN: Prospective unblinded study of physiologic effect of intervention. SETTING: Tertiary referral center. PATIENTS: Patients with SCD syndrome documented by history, sound- or pressure-evoked eye movements, vestibular-evoked myogenic potential testing, and high-resolution multiplanar computed tomographic scans. INTERVENTION: Nineteen subjects with SCD had quantitative measurements of their angular vestibulo-ocular reflexes (AVOR) in response to rapid rotary head thrusts measured by magnetic search coil technique before and after middle fossa approach and repair of the dehiscence. In 18 subjects, the dehiscence was plugged; and in 1, it was resurfaced. MAIN OUTCOME MEASURES: The AVOR gains (eye velocity/head velocity) for excitation of each of the semicircular canals. RESULTS: Vertigo resulting from pressure or loud sounds resolved in each case. Before surgery, mean AVOR gains were normal for the ipsilateral horizontal (0.94 +/- 0.07) and posterior (0.84 +/- 0.09) canals. For the superior canal to be operated on, AVOR gain was 0.75 +/- 0.13; but this was not significantly lower than the gain for the contralateral superior canal (0.82 +/- 0.11, p = 0.08). Mean AVOR gain decreased by 44% for the operated superior canals (to 0.42 +/- 0.11, p < 0.0001). There was a 13% decrease in gain for the ipsilateral posterior canal (p = 0.02), perhaps because plugging affected the common crus in some cases. There was a 10% decrease in gain for excitation of the contralateral posterior canal (p < 0.0001), which likely reflects the loss of the inhibitory contribution of the plugged superior canal during head thrusts exciting the contralateral posterior canal. Mean AVOR gain did not change for any of the other canals, but two subjects did develop hypofunction of all three ipsilateral canals postoperatively. CONCLUSION: Middle fossa craniotomy and repair of SCD reduce the function of the operated superior canal but typically preserve the function of the other ipsilateral semicircular canals. PMID- 17414043 TI - Conscientiousness in patients with Meniere's disease. AB - OBJECTIVE: To assess whether a conscientious personality profile is more prevalent in patients with Meniere's disease (MD) in comparison with other vertigo patients. Second, to investigate a sex effect and compare the obtained conscientiousness scores with published normative values. PATIENTS: A total of 108 patients were included and subdivided into one of five pathology groups: MD, benign paroxysmal positional vertigo, vestibular neuritis, vestibular schwannoma, and posttraumatic nonbenign paroxysmal positional vertigo. INTERVENTION: The conscientiousness domain of the self-reporting NEO Five-Factor Inventory questionnaire. MAIN OUTCOME MEASURE: Conscientiousness score. RESULTS: There was no significant difference between the diagnostic groups or a sex effect. MD patients had a higher score than published normal values. CONCLUSION: As assessed with the NEO Five-Factor Inventory questionnaire, MD patients do not demonstrate a higher conscientious personality profile in comparison with other vertigo patients. Therefore, this limits the clinical use of the personality profile to discriminate between patient groups. PMID- 17414044 TI - Intracanalicular facial nerve schwannoma. AB - OBJECTIVE: To investigate the capability of preoperative evaluations in predicting the precise anatomic origin of intracanalicular tumors. STUDY DESIGN: We conducted a retrospective case review. SETTING: Tertiary referral center. PATIENTS: A total of 11 patients (8 men and 3 women, 26-70 years old) with intracanalicular tumors who were treated surgically were included. INTERVENTIONS: Pure-tone average, auditory brainstem response, caloric test, and temporal magnetic resonance imaging were done in all 11 patients. Electroneuronography has only been performed since 2003, and only 3 patients were evaluated. MAIN OUTCOME MEASURE: Preoperative symptoms, pure-tone average, auditory brainstem response, caloric test, electroneuronography, and magnetic resonance imaging were compared between patients with facial nerve schwannomas and patients with vestibular schwannomas. RESULTS: Postoperatively, facial nerve schwannomas were diagnosed pathologically in 2 (18%) of 11 patients. There were not any clues suggesting facial nerve schwannoma in preoperative evaluations. CONCLUSION: A facial nerve schwannoma may be misdiagnosed as a vestibular schwannoma, especially when the tumor is confined to the internal auditory canal. There are no useful preoperative evaluation tools in predicting the precise nerve origin of intracanalicular tumors. These emphasize the need to fully inform the patient preoperatively. PMID- 17414045 TI - Prevention and treatment of cerebrospinal fluid leak after translabyrinthine acoustic tumor removal. AB - OBJECTIVE: To determine the incidence rate of cerebrospinal fluid (CSF) leak after translabyrinthine acoustic tumor removal using titanium mesh cranioplasty and to compare with previous series and historical controls. STUDY DESIGN: Database analysis with historical controls. SETTING: Tertiary referral neurotologic private practice. PATIENTS: The series of 389 patients who underwent titanium mesh cranioplasty after translabyrinthine tumor removal between March 2003 and July 2005. The results were compared with those in a group of 1,195 translabyrinthine tumor removal patients from our previously published series and with those in a group of 324 patients from the immediately preceding two-year period. INTERVENTION: Cranioplasty using titanium mesh after acoustic tumor removal. MAIN OUTCOME MEASURES: Rates of CSF leak for this method and previous methods of closure. RESULTS: Thirteen patients (3.3%) had CSF leaks when using the new method of titanium mesh closure. This compares with rates of 10.9% and 8.7% in series in which other methods of closure were used (p < 0.001 and 0.003). The rates of CSF leak requiring reoperation were 0.5%, 2.5%, and 1.9% for the new and the two older series, respectively. CONCLUSION: In our hands, titanium mesh cranioplasty seems to reduce the rate of CSF leaks after the translabyrinthine removal of acoustic tumors. PMID- 17414046 TI - Facial paralysis associated with cholesteatoma: a review of 13 cases. AB - OBJECTIVE: Report the clinical presentation and outcome of the cases of 13 patients with facial paralysis and acquired middle ear and mastoid cholesteatoma. PATIENTS: Patients with acquired cholesteatoma of the middle ear presenting with facial paralysis. INTERVENTIONS: Surgical treatment of the cholesteatoma and decompression of the facial nerve. MAIN OUTCOME MEASURES: The type and the timing of surgery, the intraoperative findings, and the postoperative facial nerve results were analyzed and related to the preoperative facial nerve function. RESULTS: All patients treated less than 7 days after the onset of the paralysis showed a normal facial function at long-term follow-up; patients who were operated on 7 days or more after the onset of paralysis showed a variable outcome. Statistical analysis showed a bigger number of House-Brackmann grade I in patients operated on within the first week of paralysis (p = 0.031). CONCLUSION: The prognosis of facial paralysis is related to the time of intervention. Early diagnosis and treatment of cholesteatoma represent, however, the mainstay of treatment. PMID- 17414047 TI - Valacyclovir and prednisolone treatment for Bell's palsy: a multicenter, randomized, placebo-controlled study. AB - OBJECTIVE: To investigate the effects of valacyclovir and prednisolone in comparison with those of placebo and prednisolone for the treatment of Bell's palsy, excluding zoster sine herpete. STUDY DESIGN: Prospective, multicenter, randomized placebo-controlled study. SETTING: Six academic tertiary referral centers. PATIENTS: Ultimately, 221 patients with Bell's palsy who were treated within 7 days of the onset. Serological and polymerase chain reaction examinations were performed to distinguish Bell's palsy from zoster sine herpete. INTERVENTION: The patients were treated with either valacyclovir (dosage, 1,000 mg/d for 5 days) plus prednisolone (VP [n = 114]) or placebo plus prednisolone (PP [n = 107]) administered orally. MAIN OUTCOME MEASURE: Recovery from the palsy was defined as a score higher than 36 using Yanagihara 40-point scoring system without facial contracture or synkinesis. The patients were followed up until complete recovery occurred or for more than 6 months in cases with a poor prognosis. RESULTS: The overall rate of patient recovery among those treated with VP (96.5%) was significantly better (p < 0.05) than the rate among those treated with PP (89.7%). The rate of patient recovery was also analyzed by classifying the initial severity of facial palsy. In cases of complete or severe palsy, the rates of patients treated with VP and PP who recovered were 95.7% (n = 92) and 86.6% (n = 82), respectively; the recovery rate for treatment with VP was significantly better than that with PP (p < 0.05). CONCLUSION: The valacyclovir and prednisolone therapy was more effective in treating Bell's palsy, excluding zoster sine herpete, than the conventional prednisolone therapy. To our knowledge, this is the first controlled study of an antiviral agent in the treatment of a sufficient number of Bell's palsy cases based on an etiologic background. PMID- 17414048 TI - Three dimensional computed tomography angiography in imaging jugular foramen lesions. PMID- 17414049 TI - The diagnostic criteria for autoimmune chronic pancreatitis: it is time to make a consensus. AB - Autoimmune chronic pancreatitis (AIP) is increasingly being recognized as a worldwide entity. In 2002, the Japan Pancreas Society published diagnostic criteria for AIP. Since then, increased attention toward this relatively new disease entity has enabled more cases of AIP to be correctly diagnosed, allowing for proper management and avoidance of surgery. Retrospective inclusion of previously unrecognized or misdiagnosed cases of AIP has revealed an increasing number of cases that are not in full accordance with the Japanese diagnostic criteria. As a result, some groups have developed and cited their own criteria in the reporting of AIP, and the Japan Pancreas Society criteria have also undergone revision recently. The absence of consistent and uniform criteria has made the comparison of different cases diagnosed under various guidelines difficult. In this review, we discuss and compare the 4 current diagnostic criteria, focusing on their own strength and weakness with the aim of providing a framework for the development of unified criteria that represent an international consensus. PMID- 17414050 TI - Endoscopic stent therapy for patients with chronic pancreatitis: results from a prospective follow-up study. AB - OBJECTIVES: Obstruction of the pancreatic duct is a common feature of chronic pancreatitis and often requires interventional therapy. The current prospective study investigated clinical success in 19 patients after initial endoscopic retrograde pancreaticography and relapse rates during a 2-year follow-up period. METHODS: Seventeen of 19 patients with chronic pancreatitis (stage III according to the Cambridge classification) were treated by sphincterotomy and stent insertion. Endoscopic retrograde pancreaticography failed in 2 patients. RESULTS: Strictures were cannulated, dilated, and stones were removed with a dormia basket in 13 of 17 patients. Extracorporeal shock wave lithotripsy was necessary in 5 patients. Polyethylene stents (7F-11.5F) were placed into the dilated pancreatic duct. Mean duration of internal pancreatic stenting was 5.6 months. Three of 17 patients had recurrence of pain during the first follow-up year after stent extraction; in the second follow-up year, another 2 patients had a relapse. Overall, patients' assessment of the stent therapy revealed complete satisfaction in 17 of 19 patients. CONCLUSIONS: Endoscopic stent therapy is a safe, minimally invasive, and effective procedure in patients experiencing pain attacks during chronic pancreatitis associated with dilated pancreatic duct. According to our results, a relapse rate of approximately 30% can be expected within 2 years after stent extraction. These patients may be treated by repeated stent therapy. PMID- 17414051 TI - Toll-like receptor 4 Asp299Gly polymorphism is associated with an increased risk of pancreatic necrotic infection in acute pancreatitis: a study in the Chinese population. AB - OBJECTIVES: To detect the frequency of Toll-like receptor (TLR) 4896A>G (Asp299Gly) polymorphism in acute pancreatitis (AP) patients and to determine the relationship between polymorphism and pancreatic necrotic infection. METHODS: The allele frequency of TLR4 Asp299Gly was surveyed by mispairing polymerase chain reaction-restriction fragment length polymorphism analysis technique in AP patients with pancreatic necrosis and healthy volunteers, respectively. RESULTS: All individuals with TLR4 Asp299Gly polymorphism were heterozygotes. Compared with healthy volunteers (5.6%), TLR4 896G allele frequencies was significantly higher in AP patients with pancreatic infection (20%), the incidence of gram negative infection was significantly higher in AP patients with TLR4 Asp299Gly polymorphism (15 [44%]/34) than that in AP patients without TLR4 polymorphism (15 [18%]/81). CONCLUSIONS: Toll-like receptor 4 Asp299Gly polymorphism is associated with the infection of pancreatic necrosis in AP. Patients who carried TLR4 896A>G mutation is susceptible to pancreatic gram-negative bacteria infection. PMID- 17414052 TI - IS-741 attenuates local migration of monocytes and subsequent pancreatic fibrosis in experimental chronic pancreatitis induced by dibutyltin dichloride in rats. AB - OBJECTIVES: Chronic pancreatitis consists of excessive leukocyte infiltration and fibrosis. IS-741 has been reported to be an antiinflammatory drug through an inhibitory action on cell adhesion. In this study, we investigated whether IS-741 could inhibit the progression of pancreatic fibrosis through monocyte infiltration. Moreover, we investigated the effect of IS-741 on rat pancreatic stellate cells (PSCs). METHODS: Chronic pancreatitis was induced by dibutyltin dichloride in rats. From days 7 to 28 after dibutyltin dichloride application, IS 741 or distilled water was administered. At days 14 and 28, histological [hematoxylin-eosin stain and immunostain for ED1 and [alpha] smooth muscle actin (alpha-SMA)] and biochemical evaluations (intrapancreatic amylase, protein, cytokines, chemokines, and alpha-SMA) were performed. In vitro, rat PSCs were incubated with cytokine, chemokine, and growth factor simultaneously with IS-741, and their proliferation and activation were examined. RESULTS: Histologically, IS 741 inhibited pancreatic fibrosis and decreased the number of ED1- and [alpha] SMA-positive cells. The intrapancreatic expression of cytokines, chemokine, and [alpha]-SMA were also decreased. In vitro, IS-741 has no direct effect on the proliferation, alpha-SMA expression, and collagen synthesis of PSCs. CONCLUSIONS: These results suggest that IS-741 suppressed macrophage infiltration and subsequent pancreatic fibrosis and that the infiltration of monocytes into pancreas is essential for pancreatic fibrosis. PMID- 17414053 TI - Plasma proteomic analysis of pancreatic cancer by 2-dimensional gel electrophoresis. AB - OBJECTIVE: Late diagnosis is the major reason for poor prognosis of pancreatic cancer (PC). Developing new biomarkers of early stage detection is critical. METHODS: Proteomic analysis with 2-dimensional gel electrophoresis was used to identify differentially expressed proteins in plasma of PC patients. The 2 dimensional gel electrophoresis plasma protein profiles of 11 PC patients (preoperative and postoperative) were compared with those of 10 patients with chronic pancreatitis (CP) and 11 healthy controls. RESULTS: Five proteins were found to be constantly changed. Haptoglobin (Hp) beta chain and leucine-rich alpha2 glycoprotein up-regulated slightly in PC plasma. Pancreatic cancer had a higher frequency of Hp2-2 phenotype but lacked Hp1-1 phenotype. Hemoglobin was increased significantly in plasma samples of PC and CP. Alpha1 antitrypsin gradually increased its expression level in healthy control, PC, and CP. Immunoglobin J chain was elevated in CP plasma samples. Haptoglobin, alpha1 antitrypsin, and leucine-rich alpha2-glycoprotein were all greatly elevated after tumor resection in PC patients. CONCLUSIONS: Proteomic analysis can simultaneously detect changes of multiproteins in plasma of PC, but detected proteins are abundant and common plasma proteins and their diagnostic value may be limited. PMID- 17414054 TI - Diagnostic and prognostic value of plasma tumor M2 pyruvate kinase in periampullary cancer: evidence for a novel biological marker of adverse prognosis. AB - OBJECTIVE: This prospective study examines the diagnostic and prognostic use of tumor-M2-pyruvate kinase (Tu-M2-PK) used in conjunction with carbohydrate antigen (CA) 19-9 in patients with subsequently histologically confirmed periampullary malignancy. METHODS: Plasma Tu-M2-PK and serum CA 19-9 levels were measured at admission in a cohort of patients with suspected pancreatic cancer. Values for Tu M2-PK and serum CA 19-9 were compared with a control group comprising jaundiced patients in whom malignancy was excluded by endoscopic retrograde cholangiopancreatography and nonjaundiced individuals undergoing laparoscopic cholecystectomy. RESULTS: The mean (SD) plasma Tu-M2-PK level for patients with histologically proven malignancy was 40.5 (26.4) U/mL and for noncancer patients, 29.9 (20.9) U/mL (Mann-Whitney U = 1163, P = 0.006). Tumor-M2-pyruvate kinase had an area under the curve of 0.623 on receiver operating characteristic curve analysis, and at optimal cutoff of 27 U/mL, sensitivity is 66%, and specificity is 58%.However, on multivariate Cox regression modeling, elevated Tu-M2-PK (>27 U/mL) was strongly correlated with the subsequent finding of poorly differentiated cancer and/or metastatic disease and strongly predicted survival on Kaplan-Meier analysis. CONCLUSION: An elevated Tu-M2-PK more than 27 U/mL measured on admission in suspected periampullary cancer is a predictor of adverse prognosis in periampullary cancer. PMID- 17414055 TI - Gene expression profiling in lymph node-positive and lymph node-negative pancreatic cancer. AB - OBJECTIVES: The purpose of this study was to screen for genes related to lymph node metastasis by comparing the differences in the expression profile between pancreatic cancer with lymph node metastasis and one without, and to predict the invasiveness and the progression of pancreatic cancer on the basis of these findings. METHODS: The total RNA of the tissues was extracted from 10 pancreatic cancer specimens, including those with lymph node metastasis and those with no metastasis. It was studied by means of a DNA microarray (oligo chip) consisting of 37,842 genes. We screened out 1.5-fold or more differential gene expressions in at least 5 pairs of samples. We classified both samples and genes using a 2 way clustering analysis. The screened-out genes were identified using real-time polymerase chain reaction. RESULTS: We identified 194 genes, including 66 overexpressed and 128 underexpressed genes, in pancreatic cancer with lymph node metastasis. Among them, we identified some genes related to lymph node metastasis in patients with pancreatic cancer: oncogenes, tumor suppressor genes, apoptosis and antiapoptosis genes, tumor angiogenesis factors, and cell cycle regulators. Genes promoting the growth of tumor cells were highly expressed in lymph node positive pancreatic cancer, whereas genes inducing apoptosis were underexpressed. CONCLUSIONS: We have identified genes that may play an important role in metastasis and survival in patients with pancreatic cancer. These results provide new insight into the study of human pancreatic cancer metastasis, including lymph node metastasis, and ultimately may lead to improving early diagnosis and discovering innovative therapeutic approaches for cancer. PMID- 17414056 TI - Impact of gemcitabine on the survival of patients with stage IV pancreatic cancer. AB - OBJECTIVES: The prognosis of patients with advanced pancreatic cancer remains very poor. This study was designed to elucidate the prognostic factors of patients with pancreatic cancer to evaluate appropriate treatment with gemcitabine. METHODS: Ninety-nine consecutive patients with stage IV pancreatic cancer were treated in the gemcitabine era at the Kobe University Hospital. Prognostic variables for survival were analyzed (sex, age, performance status, main site of the tumor, tumor size, major vessel invasion, distal metastasis, resection, gemcitabine, radiation, and pathological factors). The Cox proportional hazards model was used to determine the factors influencing the survival of patients with stage IV pancreatic cancer. RESULTS: Multivariate analysis revealed that pancreatic resection, gemcitabine, and distant metastasis significantly influenced the survival of all patients with stage IV pancreatic cancer. Pancreatic resection and gemcitabine were significant factors influencing the survival of patients with stage IVa pancreatic cancer, whereas gemcitabine was the strongest factor influencing stage IVb pancreatic cancer. CONCLUSIONS: Gemcitabine has a possible role for stage IV pancreatic cancer. PMID- 17414057 TI - Sonic hedgehog signaling pathway in pancreatic cystic neoplasms and ductal adenocarcinoma. AB - OBJECTIVES: Hedgehog (Hh) signaling is an important mediator of tumorigenesis of pancreatic ductal adenocarcinoma (PA). It is intriguing to explore whether Hh signaling is also involved in pancreatic cystic neoplasms, which are phenotypically different from PA. METHODS: Patients with solid and pseudopapillary tumor (SPT; n = 12), mucinous cystic neoplasm (MCN; n = 18), intraductal papillary mucinous neoplasm (IPMN; n = 18), and PA (n = 20) were studied. Expression of Hh signaling molecules including sonic Hh (sHh), smoothened (Smo), patched 1 (Ptc1), and Gli were determined using immunohistochemistry and/or Western blotting. Cell cycle-regulator genes, including cyclin A, B, C, and D1 messenger RNA, were determined using ribonuclease protection assay. RESULTS: Six of 12 SPT, 8 of 18 MCN, 17 of 18 IPMN, and 20 of 20 PA displayed Hh signaling using immunohistochemistry. Sonic Hh was predominantly expressed in stromal cells neighboring to the neoplastic cells of SPT and IPMN; in contrast, sHh was expressed in both stromal cells and neoplastic epithelial cells of MCN and PA. The quantitative expression of sHh signaling detected by Western blotting showed that expression of Ptc1 and Gli, but not Smo, corresponded to the magnitude of sonic hedgehog ligand. The expression of cyclin D1 messenger RNA was highest in PA, followed by MCN, IPMN, and SPT, which matches with Ptc1 and Gli. CONCLUSIONS: Hedgehog signaling pathway might play a role during tumorigenesis of SPT, MCN, IPMN, and PA. Mucinous cystic neoplasm and PA exhibit an autocrine regulation of sHh, whereas SPT and IPMN do not. Overexpression of Ptc1 and Gli, reflected by cyclin D1, might represent proliferative potential of various pancreatic neoplasms. PMID- 17414058 TI - Compromised beta-cell development and beta-cell dysfunction in weanling offspring from dams maintained on a high-fat diet during gestation. AB - OBJECTIVES: Reported here are the effects of a high-fat diet (HFD) fed to dams during pregnancy on the weight, beta- and alpha-cell development, and beta-cell function of their weanling offspring. METHODS: Offspring were obtained from dams maintained on an HFD for the first, second, or third week of gestation or throughout gestation and then on a standard laboratory diet for the duration of lactation. Weanling weights and circulating glucose and insulin concentrations were measured on postnatal day 21, after which pancreata were excised and snap frozen for quantitative polymerase chain reaction of glucokinase (GK) or processed for immunohistochemical examination and image analysis (beta- and alpha cell volume, number, and size, and GK immunoreactivity). RESULTS: All of the weanlings had low body weights and were hypoinsulinemic. In weanlings maintained on an HFD for either the first, second, or third week of gestation, hyperglycemia and a reduction in beta-cell volume and number, in beta- and alpha-cell size, and in both GK messenger RNA expression and immunoreactivity were observed. The development of beta and alpha cells was normal in weanlings maintained on an HFD throughout gestation. CONCLUSIONS: Maintenance of dams on an HFD for any single week of gestation results in weanling offspring with an impairment in beta-cell development and function. PMID- 17414059 TI - Effect of insufficient insulin treatment in streptozotocin-induced diabetes mellitus. AB - OBJECTIVES: In this study, we investigated clinically, pathologically, and immunohistochemically the effect of insufficient short-acting insulin treatment on streptozotocin (STZ)-induced diabetes mellitus in rats. METHODS: Three groups composed of 10 rats each were studied as follows: (1) a group that received only STZ (50 mg/kg) (STZ group); (2) a group that received 50 mg/kg STZ and, after 12 hours, 8 IU of short-acting insulin treatment (STZ + INS group), repeated every night for 5 days; and (3) a control group. Ketonuria and blood glucose levels were examined every day. Blood was obtained from 2 rats from each group, and necropsy was performed every day during the 5-day period. RESULTS: Hyperglycemia was observed in the STZ and STZ + INS groups 24 hours after, but levels were higher in the STS + INS group than those in the STZ-only group. Histopathology was similar in the STZ and STZ + INS groups, and degeneration was observed in both groups, but immunohistochemistry revealed a more severe reduction in insulin secreting cells in the STZ + INS group than that in the STZ group. There were no hyperglycemia and histopathological or immunochemical alteration in the control group. CONCLUSIONS: This study showed that insufficient short-acting insulin treatment can increase the diabetogenic effect of STZ in rats. PMID- 17414060 TI - Effects of palmitate on insulin secretion and exocytotic proteins in islets of diabetic Goto-Kakizaki rats. AB - OBJECTIVES: We examined how lipotoxicity contributes to pancreatic beta-cell secretory dysfunction. METHODS: Effects of palmitate (0.2 mmol/L) were assessed on insulin secretion and soluble N-ethylmaleimide-sensitive factor attachment protein receptor exocytotic machinery in isolated pancreatic islets of type 2 diabetic Goto-Kakizaki (GK) rats and control Wistar (W) rats. RESULTS: One-day palmitate treatment enhanced basal glucose (3.3 mmol/L)-mediated insulin release 5-fold in W and 3.3-fold in GK islets, but had no effect at high glucose (16.7 mmol/L) on W islets while enhancing GK islet insulin release 2-fold. After 3-day palmitate treatment, high-glucose-induced insulin release in W islets was reduced (by 69%), whereas in GK islets, it increased 2-fold. Insulin response to arginine was reduced in both islet types, but more so in GK islets. Exocytotic proteins (syntaxin 1A, VAMP-2, SNAP-25, nSec1) were reduced in GK islets by 56% to 69% compared with W islets. In W islets, palmitate treatment caused no changes in the levels of these proteins but increased actin levels. In GK islets, whereas 1-day palmitate treatment had no effect, 3-day treatment further reduced SNAP-25 and nSec1 levels. CONCLUSIONS: Lipotoxic-induced secretory insufficiency in normal islets may be attributed to lack of compensatory increase in levels of exocytotic proteins and/or excess actin. However, in GK islets, palmitate treatment moderately enhanced insulin secretion, likely by acting on proximal metabolic pathways capable of compensating for the defective soluble N-ethylmaleimide sensitive factor attachment protein receptor exocytotic machinery. These results were different from prolonged glucose treatment we previously reported, indicating differences between glucotoxic and lipotoxic actions on the insulin secretory machinery. PMID- 17414061 TI - High glucose activates rat pancreatic stellate cells through protein kinase C and p38 mitogen-activated protein kinase pathway. AB - OBJECTIVE: Hyperglycemia is implicated in fibrosis in many organs. Exocrine and endocrine pancreas are closely linked both anatomically and physiologically, and pathological conditions in the exocrine gland can cause impairment of endocrine function and vice versa. Chronic pancreatitis causes pancreatic fibrosis and sometimes results in diabetes mellitus. Pancreatic stellate cells (PSCs) play a pivotal role in pancreatic fibrogenesis. However, the effects of high glucose concentrations on PSC activation have not been fully elucidated. METHODS: Cultured PSCs were incubated in the presence of various concentrations of glucose. Pancreatic stellate cell proliferation, alpha-smooth muscle actin (alpha SMA) expression, and collagen production were determined by colorimetric conversion assay, Western blot analysis, and Sirius red dye binding assay, respectively. RESULTS: High glucose concentrations significantly increased PSC proliferation, alpha-SMA expression, and collagen type I production in PSCs. High glucose concentrations activated protein kinase C (PKC) in PSCs, and PKC inhibitor GF109203X inhibited glucose-stimulated PSC proliferation, alpha-SMA expression, and collagen secretion. High glucose also activated p38 mitogen activated protein kinase (MAPK) in PSCs, and p38 MAPK inhibitor SB203580 inhibited glucose-stimulated collagen secretion. CONCLUSIONS: Our results indicate that high glucose concentrations stimulate PSC activation via PKC-p38 MAP kinase pathway and suggest that high glucose may aggravate pancreatic fibrosis. PMID- 17414062 TI - Effects of gonadal steroid hormones on amylase secretion in pancreatic lobules of rats. AB - OBJECTIVES: Because functional changes of the pancreas during the ovarian cycle are not fully understood, effects of gonadal steroid hormones on pancreatic amylase content and secretion were investigated. METHODS: The estrus cycle of female rats was confirmed by vaginal smear. To mimic the estrus or the diestrus, estradiol 17beta (25 microg/kg) or progesterone (50 mg/kg) was subcutaneously injected to ovariectomized rats once daily for 2 days. Amylase secretion of pancreatic lobules (approximately 6 mg wet weight) was induced by acetylcholine (10-8 approximately 10-4 M). RESULTS: In normal rats, pancreatic amylase content was not altered during the estrus cycle, whereas spontaneous amylase secretion of pancreatic lobules at the diestrus was significantly higher than that at the estrus. In ovariectomized rat, pancreatic amylase content was markedly reduced, which was restored by either estradiol 17beta or progesterone. Pancreatic lobules of ovariectomized rats spontaneously secreted amylase at the level identical to that at the estrus, which was elevated to the level at the diestrus by progesterone, but not affected by estradiol 17beta. In normal rats, acetylcholine induced amylase secretion much higher at the diestrus than at the estrus. In ovariectomized rats, the acetylcholine-induced amylase secretion was similar to that at the estrus, which was elevated by progesterone, but not affected by estradiol 17beta. CONCLUSION: We conclude from the above results that both estradiol 17beta and progesterone are necessary to maintain amylase content in the rat pancreas, but only progesterone exerts a stimulatory effect on spontaneous and stimulated amylase secretion in pancreatic lobules of rats. PMID- 17414063 TI - Acute tubulointerstitial nephritis associated with long-term gemcitabine therapy. PMID- 17414064 TI - Propoxur induced pancreatitis after inhalation of baygon pesticide. PMID- 17414065 TI - Pancreatitis due to catastrophic antiphospholipid syndrome. PMID- 17414066 TI - A novel procedure for the management of pancreatic anastomotic insufficiency. PMID- 17414067 TI - Partner notification for gonorrhea-time for new ideas. PMID- 17414068 TI - Universal screening or prophylactic treatment for Chlamydia trachomatis infection among women seeking induced abortions: which strategy is more cost-effective? AB - OBJECTIVE: The objective of this study was to evaluate the cost-effectiveness of universal screening and azithromycin-based prophylaxis against no intervention for Chlamydia trachomatis infection among women seeking induced abortions. METHODS: A decision tree was constructed to evaluate health effects of the program. Cost-effectiveness was estimated for universal screening and azithromycin-based prophylaxis against no intervention with a C. trachomatis test prevalence of 4.8%. RESULTS: Azithromycin-based prophylaxis produced higher cost but prevented 289 cases of pelvic inflammatory disease (PID) for a cost of 397 RMB (U.S. $48) per case of PID prevented over no intervention. Universal screening by polymerase chain reaction test prevented 253 cases of PID at a cost of 3,049 RMB (U.S. $372) per case of PID prevented over no intervention. Azithromycin-based prophylaxis prevented an additional 36 cases of PID, costing 18,239 RMB (US $2,224) less per case of PID prevented over universal screening. CONCLUSIONS: Azithromycin-based prophylaxis provided a cost savings over universal screening for chlamydial infection among women seeking induced abortion. PMID- 17414069 TI - Sexually transmitted infections and sexual practices in injection drug users and their regular sex partners in Chennai, India. AB - OBJECTIVE: Measuring sexually transmitted infections (STIs) and sexual practices in injection drug users (IDUs) and their regular sex partners. GOAL: Informing HIV intervention programs. DESIGN: Cross-sectional. RESULTS: One percent IDUs and 2% of their regular female sex partners were syphilis infected; 40% (84/211) and 38% respectively (81/211) were infected with HSV-2. 30% IDUs and 5% of their female regular sex partners were HIV positive. Serodiscordant results for syphilis and HSV-2 were noticed. Women having first sex at ageor=38 years had seven times the odds of having any non-HIV-STI. CONCLUSION: Reaching out to IDUs and their female regular sex partners with modified STI management guideline and promoting women-controlled safer sex measures are needed harm-reduction measures. PMID- 17414071 TI - Image-based control of the magnetic resonance imaging-guided focused ultrasound thermotherapy. AB - Magnetic resonance imaging (MRI)-guided focused ultrasound surgery (FUS) is a full noninvasive approach for localized thermal ablation of deep tissues, coupling the following: (1) a versatile, nonionizing physical agent for therapy and (2) a state-of-the art diagnosis and on-line monitoring tool. A commercially available, Food and Drug Administration-approved device using the MRI-guided FUS exists since 2004 for the ablation of benign tumors (uterine fibroids); however, the ultimate goal of the technological, methodological, and medical research in this field is to provide a clinical-routine tool for fighting localized cancer. When addressing cancer applications, the accurate spatial control of the delivered thermal dose is mandatory. Contiguous destruction of the target volume must be achieved in a minimum time, whereas sparing as much as possible the neighboring healthy tissues and especially when some adjacent regions are critical. This paper reviews some significant developments reported in the literature related to the image-based control of the FUS therapy for kidney, breast, prostate, and brain, including the own experience of the authors on the active feedback control of the temperature during FUS ablation. In addition, preliminary results of an original study of MRI-guided FUS ablation of VX2 carcinoma in kidney, under active temperature control, are described here. PMID- 17414072 TI - Magnetic resonance temperature imaging for focused ultrasound surgery: a review. AB - Magnetic resonance temperature imaging (MRTI) is an enabling technology that has recently demonstrated the potential to bring the emerging minimally invasive image-guided thermal therapy procedures, such as radiofrequency, microwave, laser, ultrasound, and cryosurgery, into the clinical setting with a level of safety and efficacy not previously possible. By coupling the wealth of soft tissue contrast mechanisms available with magnetic resonance imaging with its intrinsic temperature sensitivity, magnetic resonance imaging is in a unique position to provide image-guided treatment planning and verification and quantitative or qualitative feedback during treatment delivery, heightening of the control the physician has over the method, and enhancement of the ability to deliver conformal treatments. The basic principles behind MRTI technology and its application to minimally invasive thermal therapy during ultrasound thermal therapy delivery are reviewed in this study. PMID- 17414073 TI - Transcranial focused ultrasound surgery. AB - The development of high-intensity ultrasound technology into a system for performing image-guided noninvasive ultrasound neurosurgery has developed at a relatively rapid pace in the past few years. Magnetic resonance imaging has contributed significantly to this development by providing a modality by which percutaneous ultrasound treatments can be preoperatively planned, intraoperatively guided and postoperatively evaluated for safety and efficacy. Especially in the case of transcranial ultrasound therapies, the structural identification and thermal monitoring of cortical structures is essential to avoid overheating at the skull-brain interface and to avoid the sonication of critical structures. This chapter briefly describes the physics of transmitting ultrasound through the skull and the technological advances that circumvented the physical limits imposed by the skull bone. The integration of magnetic resonance guidance and monitoring is detailed, along with an overview of ongoing studies with a commercially developed magnetic resonance imaging-compatible hemispherical transducer array. PMID- 17414074 TI - A review of magnetic resonance imaging-guided focused ultrasound surgery of uterine fibroids. AB - Uterine fibroids are a significant cause of morbidity for women of reproductive age. Over the past decade, minimally invasive treatment options are becoming increasingly popular. A new, Food and Drug Administration-approved noninvasive treatment option is magnetic resonance-guided focused ultrasound surgery, which has the potential to become a treatment of choice for selected patients. We review the technical aspects of the procedure of magnetic resonance-guided focused ultrasound surgery for treatment of uterine fibroids, potential difficulties with treatment planning, and clinical trial results to date. We also describe current developments in treatment imaging and treatment optimization. PMID- 17414075 TI - Breast focused ultrasound surgery with magnetic resonance guidance. AB - This paper will review the experience and current applications of magnetic resonance-guided focused ultrasound surgery (MRgFUS) for treatment of breast tumors. Because of the efficient screening mammography programs, most of the breast cancers diagnosed today in the United States and European Union are in early stage and are treated with limited surgery. The MRgFUS may offer an alternative treatment option to conventional surgical lumpectomy with the advantage of being a noninvasive procedure and potentially achieving a better cosmetic outcome. Selection of appropriate patients is of paramount importance. Additional studies are needed to determine the effectiveness of the MRgFUS tumor ablation and define its role as a replacement for surgical lumpectomy. PMID- 17414076 TI - New clinical applications of magnetic resonance-guided focused ultrasound. AB - Magnetic resonance-guided ultrasound delivers destructive energy into deep body structures with great accuracy and repeatability with an excellent safety profile. The use of this technology for the treatment of uterine fibroids is already becoming widespread. This article reviews the further areas of magnetic resonance-guided focused ultrasound application that are evolving and how they will be applied to other parts of the body. PMID- 17414077 TI - Magnetic resonance-guided high-intensity ultrasound ablation of the prostate. AB - OBJECTIVES: This paper describes our work in developing techniques and devices for magnetic resonance (MR)-guided high-intensity ultrasound ablation of the prostate and includes review of relevant literature. METHODS: Catheter-based high intensity ultrasound applicators, in interstitial and transurethral configurations, were developed to be used under MR guidance. Magnetic resonance thermometry and the relevant characteristics and artifacts were evaluated during in vivo thermal ablation of the prostate in 10 animals. Contrast-enhanced MR imaging (MRI) and diffusion-weighted MRI were used to assess tissue damage and compared with histology. RESULTS: During evaluation of these applicators, MR thermometry was used to monitor the temperature distributions in the prostate in real time. Magnetic resonance-derived maximum temperature thresholds of 52 degrees C and thermal dose thresholds of 240 minutes were used to control the extent of treatment and qualitatively correlated well with posttreatment imaging studies and histology. The directional transurethral devices are selective in their ability to target well-defined regions of the prostate gland and can be rotated in discrete steps to conform treatment to prescribed boundaries. The curvilinear applicator is the most precise of these directional techniques. Multisectored transurethral applicators, with dynamic angular control of heating and no rotation requirements, offer a fast and less complex means of treatment with less selective contouring. CONCLUSIONS: The catheter-based ultrasound devices can produce spatially selective regions of thermal destruction in prostate. The MR thermal imaging and thermal dose maps, obtained in multiple slices through the target volume, are useful for controlling therapy delivery (rotation, power levels, duration). Contrast-enhanced T1-weighted MRI and diffusion-weighted imaging are useful tools for assessing treatment. PMID- 17414078 TI - Targeted drug delivery to the brain using focused ultrasound. AB - Drug delivery to the brain remains a challenging field. The presence of a physiological barrier, the blood-brain barrier (BBB), complicates the delivery of drugs to the brain. Although several methods have been developed for drug delivery to the brain, they have problems such as being invasive or lacking in target specificity. On the other hand, ultrasound has emerged as a treatment method and a diagnostic technology. Several studies have shown the feasibility of using ultrasound for the localized and reversible disruption of the BBB. In this review, I would like to review the recent advancement of ultrasound-induced MRI guided BBB disruption technique and other methods for delivering drugs to the brain. PMID- 17414079 TI - Rapid response systems: a systematic review. AB - CONTEXT: Rapid response systems have been advocated as a potential model to identify and intervene in patients who are experiencing deterioration on general hospital wards. OBJECTIVE: To conduct a meta-analysis to evaluate the impact of rapid response systems on hospital mortality and cardiac arrest rates. DATA SOURCE: We searched MEDLINE, EMBASE, and the Cochrane Library from January 1, 1990, to June 30, 2005, for all studies relevant to rapid response systems. We restricted the search to the English language and by age category (all adults: >or=19 years). STUDY SELECTION: We selected observational and randomized trials of rapid response systems that provided empirical data on hospital mortality and cardiac arrest in control and intervention groups. We reviewed 10,228 abstracts and identified eight relevant studies meeting these criteria. DATA SYNTHESIS: Of the included studies, five used historical controls, one used concurrent controls, and two used a cluster-randomized design. The pooled relative risk for hospital mortality comparing rapid response teams to control was 0.76 (95% confidence interval, 0.39-1.48) between the two randomized studies and 0.87 (95% confidence interval, 0.73-1.04) among the five observational studies. The pooled relative risk for cardiac arrest comparing rapid response systems to control was 0.94 (95% confidence interval, 0.79-1.13) in the single randomized study and 0.70 (95% confidence interval, 0.56-0.92) in four observational studies. CONCLUSIONS: We found weak evidence that rapid response systems are associated with a reduction in hospital mortality and cardiac arrest rates, but limitations in the quality of the original studies, the wide confidence intervals, and the presence of heterogeneity limited our ability to conclude that rapid response systems are effective interventions. Large randomized controlled trials are needed to clarify the efficacy of rapid response systems before they should become standard of care. PMID- 17414080 TI - Economic implications of an evidence-based sepsis protocol: can we improve outcomes and lower costs? AB - OBJECTIVE: To determine the financial impact of a sepsis protocol designed for use in the emergency department. DESIGN: Retrospective analysis of a before-after study testing the implications of sepsis protocol. SETTING: Academic, tertiary care hospital in the United States. PATIENTS: Persons with septic shock presenting to the emergency department. INTERVENTIONS: A multifaceted protocol developed from recent scientific literature on sepsis and the Surviving Sepsis Campaign. The protocol emphasized identification of septic patients, aggressive fluid resuscitation, timely antibiotic administration, and appropriateness of antibiotics, along with other adjunctive, supportive measures in sepsis care. MEASUREMENTS AND MAIN RESULTS: We compared patients treated before the protocol with those cared for after the protocol was implemented. Overall hospital costs represented the primary end point, whereas hospital length of stay served as a secondary end point. All hospital costs were calculated based on charges after conversion to costs based on department-specific cost-to-charge ratios. We also attempted to measure the independent impact of the protocol on costs through linear regression. We conducted a sensitivity analysis assessing these end points in the subgroup of subjects who survived their hospitalization. The total cohort included 120 subjects (evenly divided into the before and after cohorts) with a mean age of 64.7 +/- 18.2 yrs and median Acute Physiology and Chronic Health Evaluation II score of 22.5 +/- 8.3. There were more survivors following the protocol's adoption (70.0% vs. 51.7%, p = .040). Median total costs were significantly lower with use of the protocol ($16,103 vs. $21,985, p = .008). The length of stay was also on average 5 days less among the postintervention population (p = .023). A Cox proportional hazard model indicated that the protocol was independently associated with less per-patient cost. Restricting the analysis to only survivors did not appreciably change our observations. CONCLUSIONS: Use of a sepsis protocol can result not only in improved mortality but also in substantial savings for institutions and third party payers. Broader implementation of sepsis treatment protocols represents a potential means for enhancing resource use while containing costs. PMID- 17414081 TI - Relationship between immunosuppression and intensive care unit-acquired multidrug resistant bacteria: a case-control study. AB - OBJECTIVE: To determine the relationship between immunosuppression and intensive care unit (ICU)-acquired multidrug-resistant (MDR) bacteria. DESIGN: Retrospective case-control study based on prospectively collected data. SETTING: A 30-bed medical and surgical ICU. PATIENTS: All patients hospitalized >48 hrs in the ICU were eligible during a 2-yr period. INTERVENTIONS: Immunosuppression was defined as active solid or hematologic malignancy, leucopenia, or chronic immunosuppressive treatment. MDR bacteria were defined as methicillin-resistant Staphylococcus aureus, ceftazidime- or imipenem-resistant Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, and extending spectrum beta-lactamase producing Gram-negative bacilli. MDR bacteria screening (nasal, anal, and axilla swabs and tracheal aspirate in intubated patients) was performed at ICU admission and weekly. Only MDR bacteria isolated >48 hrs after ICU admission were taken into account; duplicates were excluded. Isolation measures were applied in all patients at ICU admission, in patients with MDR bacteria, and in patients with immunosuppression. Immunosuppressed patients (cases) were matched (1:1) with immunocompetent patients (controls) according to all the following criteria: age +/-5 yrs, Simplified Acute Physiology Score II +/-5, duration of ICU stay +/-3 days, and category of admission (medical/surgical). Risk factors for ICU-acquired MDR bacteria were determined using univariate and multivariate analyses. MEASUREMENTS AND MAIN RESULTS: Of 1,065 eligible patients, nine patients were excluded for absence of MDR bacteria screening at ICU admission. One hundred thirty-three (12%) patients were immunosuppressed, and 128 (96%) of them were successfully matched. Mean time between ICU admission and first ICU-acquired MDR bacteria was 12 +/- 9 days. Incidence of MDR bacteria was significantly higher in cases than in controls (22 vs. 12 MDR bacteria/1000 ICU days, p = .004). However, immunosuppression was not independently associated with ICU-acquired MDR bacteria.Multivariate analysis identified prior antibiotic treatment and antibiotic treatment in the ICU as risk factors for ICU-acquired MDR bacteria (odds ratio [95% confidence interval] = 1.9 [1-3.6], p = .003; 11 [1.4-83], p = .02; respectively). CONCLUSIONS: Immunosuppression is not independently associated with ICU-acquired MDR bacteria. However, infection control measures used in our ICU may have influenced this result. PMID- 17414082 TI - Texas hospitals' experience with the Texas Advance Directives Act. AB - OBJECTIVE: The Texas Advance Directives Act (TADA) provides legal immunity for physicians who discontinue life-sustaining treatment judged to be medically inappropriate. The process includes review and affirmation of physicians' judgments by an ethics or medical committee. This study was undertaken to determine awareness of and experience with the medical appropriateness review process at Texas Hospital Association (THA) member hospitals from 1999 to 2004. DESIGN: Cross-sectional, descriptive, 20-item written survey instrument. SETTING: University cancer center. SUBJECTS: Subjects were 409 hospital members of THA in 2004. INTERVENTIONS: Mailed surveys. MEASUREMENTS AND MAIN RESULTS: Participants returned 197 of 409 surveys usable for analysis (48.2%). Eighty-one percent of respondents (n = 159) were aware of all the provisions of the TADA. Thirty percent of respondents (n = 58) stated that their hospitals' TADA policy or procedure had been used to review specific patient cases. However, only 46 hospitals indicated a specific number of cases reviewed. Six of these 46 surveys were judged to be too inconsistent to be usable. The 40 remaining hospitals reviewed a total of 256 cases. For 70% of the 256 reported cases (n = 178), review committees agreed with physicians that the treatments in question were medically inappropriate. CONCLUSIONS: A minority of THA hospitals have used their policies or procedures to review specific patient cases. Most cases were resolved before the end of the mandated 10-day waiting period because patients died, patients or representatives agreed to forgo the treatment in question, or patients were transferred. Discontinuation of life-sustaining treatment against patient or patient representative wishes occurred in only a small number of cases. PMID- 17414083 TI - Sepsis since the discovery of Toll-like receptors: disease concepts and therapeutic opportunities. AB - OBJECTIVE: Sepsis and its sequelae are the leading cause of death in critically ill patients. Discovery in the late 1990s of Toll-like receptors as primary sensors of microbial infection led to significant advances in understanding the pathogenesis of sepsis, including emerging differences between Gram-positive and Gram-negative infection and the potential for the manipulation of Toll-like receptors for the treatment of sepsis. This review describes these advances. METHODS: Bibliographic search of the literature since 1999, with particular emphasis on the conceptual and therapeutic implications of Toll-like receptors for patients with systemic sepsis. RESULTS AND CONCLUSIONS: Toll-like receptors initiate the inflammatory processes that underlie the clinical response to infection and therefore represent an important putative target for therapeutic intervention. PMID- 17414084 TI - Is all ventricular fibrillation the same? A comparison of ischemically induced with electrically induced ventricular fibrillation in a porcine cardiac arrest and resuscitation model. AB - OBJECTIVES: The standard porcine cardiac arrest model uses electrical induction of ventricular fibrillation. Reported restoration of spontaneous circulation and survival rates in this model are as high as 90% for ventricular fibrillation durations of 7-10 mins, values substantially greater than rates in the clinical population (i.e., 20% to 30%). A high first shock success rate, infrequent refibrillation, and short times for restoration of spontaneous circulation are typical of the model. The purpose of this study was to determine whether ischemic induction of ventricular fibrillation in swine followed by standard advanced cardiac life support would result in short-term outcomes approximating those observed in human victims of out-of-hospital ventricular fibrillation. DESIGN: Randomized comparative trial. SETTING: Translational research laboratory. SUBJECTS: Domestic swine (n = 40, mean weight 40 +/- 4 kg, range 34-47 kg) of both genders. INTERVENTIONS: Swine were instrumented and randomized to either electrical ventricular fibrillation induction or ischemic ventricular fibrillation, produced by balloon occlusion of the mid-left anterior descending coronary artery (n = 20 per group). Transthoracic impedance was measured and 30 Omega added in series for all animals. The balloon remained inflated during resuscitation efforts in ischemic ventricular fibrillation animals. After 7 mins of ventricular fibrillation, cardiopulmonary resuscitation was initiated and defibrillation was attempted 1 min later. Epinephrine and antiarrhythmics were administered as per guidelines. Resuscitation was terminated if restoration of spontaneous circulation had not occurred after 15 mins of advanced cardiac life support. MEASUREMENTS AND MAIN RESULTS: Although the number of countershocks required to initially terminate ventricular fibrillation was not different (electrical ventricular fibrillation 1.9 +/- 1.6, ischemic ventricular fibrillation 2.4 +/- 2.0), the refibrillation rate was higher in the ischemic ventricular fibrillation group (4.9 +/- 4 vs. 0.8 +/- 1 episodes/animal, p < .001), resulting in a greater number of shocks before restoration of spontaneous circulation (total shocks for ischemic ventricular fibrillation 9.4 +/- 5.6 vs. electrical ventricular fibrillation 2.7 +/- 2.2, p < .001). Time to restoration of spontaneous circulation was longer in the ischemic ventricular fibrillation group (430 +/- 234 secs vs. 149 +/- 120 secs, p < .001). Restoration of spontaneous circulation rates were not different (electrical ventricular fibrillation 90% vs. ischemic ventricular fibrillation 65%). However, survival to 6 hrs was greater in the electrical ventricular fibrillation group (18 of 20, 90%) than in the ischemic ventricular fibrillation group (8 of 20, 40%, p = .002). CONCLUSIONS: Resuscitation from ischemic ventricular fibrillation is more difficult than electrical ventricular fibrillation and is characterized by greater time to restoration of spontaneous circulation, frequent refibrillation, greater number of countershocks, higher epinephrine dose during resuscitation efforts, profound cardiac dysfunction, and a short-term survival rate approaching clinical experience. Ischemically induced ventricular fibrillation is a more clinically relevant model for the evaluation of resuscitation interventions. PMID- 17414085 TI - Calculation of intratracheal airway pressure in ventilated neonatal piglets with endotracheal tube leaks. AB - OBJECTIVE: In ventilated neonates, only the applied pressure of the ventilator is adjusted and monitored. When an endotracheal tube leaks, intratracheal pressure decreases depending on the size of the endotracheal tube and of the leak. Furthermore, an increase in resistance and/or compliance might delay the increase of intratracheal pressure during inspiration and its decline during expiration. Short inspiratory time can cause insufficient ventilation, because intratracheal pressure peak might not be reached. Short expiratory time may lead to air trapping, because intratracheal pressure could not return to baseline. The aim of this study was to develop a mathematical algorithm to calculate intratracheal pressure continuously during ventilation and to evaluate the accuracy of this method. DESIGN: Prospective, animal study. SETTING: University research laboratory. SUBJECTS: To verify the mathematical algorithm, eight neonatal piglets (1600-2600 g) were studied under different endotracheal tube leak conditions (45% to 98%). The median compliance and resistance were 1.06 mL/cm H2O/kg and 123 cm H2O/L/sec, respectively. INTERVENTIONS: Pressure decreases caused by the different endotracheal tubes were measured in a model while air flow was increased stepwise. Based on these results, a mathematical method was developed to calculate intratracheal pressure under leak conditions continuously in relation to the flow through the endotracheal tube as well as to calculate the values of resistance, compliance, and applied pressure of the ventilator. MEASUREMENTS AND MAIN RESULTS: The intratracheal pressure calculated was compared with the measured intratracheal pressure over time. The differences between measured and calculated intratracheal pressure related to peak applied pressure of the ventilator did not exceed 10%. The medians of absolute amounts of differences between measured and calculated intratracheal pressure were <1 cm H2O. CONCLUSIONS: The accuracy of the calculation of intratracheal pressure ensures adequate monitoring of artificial ventilation, even in the presence of endotracheal tube leaks. This might decrease the risk of barotrauma and improve the effectiveness of ventilation. PMID- 17414086 TI - Central venous catheterization. AB - OBJECTIVE: To provide current information related to central venous catheterization. DESIGN: Review of literature relevant to central venous catheterization and its indications, insertion techniques, and prevention of complications. RESULTS: Central venous catheterization can be lifesaving but is associated with complication rates of approximately 15%. Operator experience, familiarity with the advantages and disadvantages of the various catheterization sites, and strict attention to detail during insertion help in reducing mechanical complications associated with catheterization. Strict aseptic technique and proper catheter maintenance decrease the frequency of catheter related infections. CONCLUSIONS: Appropriate catheter and site selection, sufficient operator experience, careful technique, and proper catheter maintenance with removal as soon as possible are associated with optimal outcome. PMID- 17414087 TI - Albumin resuscitation improves ventricular contractility and myocardial tissue oxygenation in rat endotoxemia. AB - OBJECTIVE: Fluid resuscitation to improve delivery of oxygen to vital organs is a principal clinical intervention for septic patients. We previously reported that albumin resuscitation in rat endotoxemia improved contractility in isolated cardiomyocytes, but whether this effect occurs in vivo is unknown. We hypothesized that albumin resuscitation would improve decreased ventricular contractility and myocardial tissue oxygenation in vivo. DESIGN: Randomized, controlled, prospective animal study. SETTING: University animal laboratory. SUBJECTS: Male Sprague-Dawley rats (250-350 g). INTERVENTIONS: Rats were randomized into three groups: control with no lipopolysaccharide (n = 8), lipopolysaccharide (10 mg/kg) without albumin resuscitation (n = 8), and lipopolysaccharide with albumin resuscitation (n = 6). Five hours after lipopolysaccharide injection, animals were resuscitated with 10 mL/kg 5% rat albumin in 0.9% saline. Six hours after 10 mL/kg lipopolysaccharide, a pressure volume conductance catheter (MIKRO-Tip 2.0-Fr, Millar Instruments, Houston, TX) was inserted into the left ventricle to quantify maximum elastance as an index of contractility. Myocardial tissue Po2 was measured using a fiberoptic oxygen probe. MEASUREMENTS AND MAIN RESULTS: Maximum elastance decreased after lipopolysaccharide relative to control (47%, from 5.9 +/- 0.8 to 3.1 +/- 0.4 mm Hg/microL, p < .05). Albumin resuscitation prevented the lipopolysaccharide induced decrease in maximum elastance (7.0 +/- 1.2 mm Hg/microL, p < .05 vs. lipopolysaccharide). Myocardial tissue Po2 was reduced in endotoxemia compared with control (53%, from 10.1 +/- 0.9 to 4.7 +/- 0.6 mm Hg, p < .05), and albumin resuscitation improved the lipopolysaccharide-induced tissue hypoxia toward the control value (9.0 +/- 1.4 mm Hg, p < .05). CONCLUSIONS: Albumin resuscitation improved decreased ventricular contractility and myocardial oxygenation in endotoxemic rats. This result suggests that albumin resuscitation may improve ventricular dysfunction by improving myocardial hypoxia. PMID- 17414088 TI - Pre-B-cell colony-enhancing factor gene polymorphisms and risk of acute respiratory distress syndrome. AB - OBJECTIVE: Pre-B-cell colony-enhancing factor (PBEF) levels are elevated in bronchoalveolar lavage fluid and serum of patients with acute lung injury. There are several suspected functional polymorphisms of the corresponding PBEF gene. We hypothesized that variations in PBEF gene polymorphisms alter the risk of developing acute respiratory distress syndrome (ARDS). DESIGN: Nested case control study. SETTING: Tertiary academic medical center. PATIENTS: We studied 375 patients with ARDS and 787 at-risk controls genotyped for the PBEF T-1001G and C-1543T polymorphisms. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients with the -1001G (variant) allele had significantly greater odds of developing ARDS than wild-type homozygotes (odds ratio, 1.35; 95% confidence interval, 1.02-1.78). Patients with the -1543T (variant) allele did not have significantly different odds of developing ARDS than wild-type homozygotes (odds ratio, 0.86; 95% confidence interval, 0.65-1.13). When analysis was stratified by ARDS risk factor, -1543T was associated with decreased odds of developing ARDS in septic shock patients (odds ratio, 0.66; 95% confidence interval, 0.45-0.97). Also, -1001G was associated with increased hazard of intensive care unit mortality, whereas -1543T was associated with decreased hazard of 28-day and 60 day ARDS mortality, as well as shorter duration of mechanical ventilation. Similar results were found in analyses of the related GC (-1001G:-1543C) and TT ( 1001T:-1543T) haplotypes. CONCLUSIONS: The PBEFT-1001G variant allele and related haplotype are associated with increased odds of developing ARDS and increased hazard of intensive care unit mortality among at-risk patients, whereas the C 1543T variant allele and related haplotype are associated with decreased odds of ARDS among patients with septic shock and better outcomes among patients with ARDS. PMID- 17414089 TI - The critically ill avian influenza A (H5N1) patient. AB - OBJECTIVE: This review examines perspectives of human infection with avian influenza A H5N1 (AI H5N1), specifically focusing on the presentation, diagnosis, and management of those critically ill with AI H5N1. DATA SOURCE: PubMed (1966 2006), PubMed "related articles," publications and Web sites of the World Health Organization and the Centers for Disease Control and Prevention, personal files, abstract proceedings, and reference lists. STUDY SELECTION: We reviewed English language publications pertaining to clinical presentation, diagnosis, and management of AI H5N1 and infection control expressly relating to the intensive care setting. DATA SYNTHESIS: The majority of reported patients with AI H5N1 are critically ill and require intensive care management. These patients progress rapidly to severe acute respiratory distress syndrome. Multiorgan failure occurs in a large proportion. Because of the nonspecific clinical, laboratory, and radiologic features, it is critical to seek a history of exposure to poultry or wild birds in suspected cases. Reverse transcription polymerase chain reaction performed on nasopharyngeal aspirate is the most reliable method for the laboratory diagnosis of AI H5N1. Treatment includes starting neuraminidase inhibitor oseltamivir as early as possible in addition to the standard supportive management. Aerosol generating procedures should be minimized to avoid nosocomial transmission. Strict infection control procedures are paramount to staff safety, although human-to-human transmission is rare as of this time. CONCLUSIONS: Many patients with AI H5N1 are critically ill either at presentation or shortly thereafter. Intensivists and intensive care units are therefore at the front line for this new cause of severe lung injury. Practitioners must be familiar with the nonspecific presentation of AI H5N1 and its diagnostic and therapeutic options. Although treating the infected patient with AI H5N1 is a priority, safeguarding healthcare workers and other patients must be considered of equal priority. PMID- 17414090 TI - Healthcare climate: a framework for measuring and improving patient safety. AB - OBJECTIVES: Reviews of patient safety efforts suggest that technical/administrative change must be augmented by global factors such as organizational culture and climate. The objective was to outline a comprehensive model for healthcare climate and test one of its elements, the nursing subclimate, in terms of several patient safety outcomes. DESIGN: Measure organizational climate in nursing units, followed by random sampling of patient safety practices in each unit 6 months later. SETTING: Sixty-nine inpatient units in three hospitals that make up the entire tertiary care system in one metropolitan area. SUBJECTS: A total of 955 nurses. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A two-part Nursing Climate Scale referring to hospital- and unit-level climates, followed by five randomly timed observations of patient safety practices covering routine and emergency care in each unit. Climate scales met the criteria of internal reliability, within-unit agreement, and between-unit variability, using standard statistics of climate research. Both the hospital and unit nursing climates exhibited significant variation, which predicted the routine medication safety scores (Z = 2.65 and 2.93 accordingly, p < .01), with similar results for emergency safety scores. A significant interaction (Z = 2.78, p < .01) indicated that best/worst safety is obtained when the unit and hospital climates are aligned (for better or worse) and that positive unit climate can compensate for the detrimental effect of poor hospital climate. Furthermore, climate's strength increased its predictive power with regard to patient safety practices (Z = 3.64 for medication and 2.28 for emergency safety; p < .01). The small number of participating hospitals limits organization-level analyses. CONCLUSIONS: The nursing climate identifies units where the likelihood of adverse events is greater or lower than the hospital's average. Such information can guide prevention efforts in selected units. These data encourage the development of additional climate subscales subsumed under the healthcare climate model (e.g., physicians subclimate). PMID- 17414091 TI - Myocardial infarction following coronary artery bypass graft surgery increases healthcare resource utilization. AB - OBJECTIVE: To assess the health economic impact of perioperative myocardial infarction in a cohort of patients undergoing coronary artery bypass graft surgery. DESIGN: Retrospective cohort analysis using data from hospital bills and uniform billing forms. SETTING: A total of 147 geographically diverse hospitals in the United States. PATIENTS: The study population consisted of 2,102 coronary artery bypass graft surgery patients enrolled in the PRIMO-CABG trial at U.S. sites between January 2002 and February 2003. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Resource utilization and costs during the index hospitalization and during a 6-month follow-up period were compared between patients who had a myocardial infarction by postoperative day 4 and those who did not. Linear regression was used to examine whether myocardial infarction by day 4 was associated with index hospitalization costs, after adjusting for baseline characteristics. Myocardial infarction occurred in 191 (9.1%) patients undergoing coronary artery bypass graft surgery. Myocardial infarction was associated with a doubling of intensive care unit time (3.5 days among patients with no myocardial infarction and 7.1 days among patients with myocardial infarction, p < .01) and a 48% increase in hospital length of stay. Myocardial infarction by day 4 was associated with a 43% increase in hospital costs, a 29% increase in physician service costs, a 41% increase in total costs during the index hospitalization, and a 38% increase in cumulative 6-month costs. After baseline adjustment, myocardial infarction continued to be associated with higher index hospitalization costs. CONCLUSIONS: Myocardial infarction following coronary artery bypass graft surgery was associated with a significant increase in intensive care unit time, hospital length of stay, and overall costs, which contributed to greater hospital and physician service costs. Healthcare resource utilization is increased in patients sustaining a myocardial infarction following coronary artery bypass graft surgery. PMID- 17414092 TI - Effects of dietary equol on body weight gain, intra-abdominal fat accumulation, plasma lipids, and glucose tolerance in ovariectomized Sprague-Dawley rats. AB - OBJECTIVE: To evaluate the effects of dietary equol, a metabolite of the phytoestrogen daidzein, on body weight gain, intra-abdominal fat accumulation, plasma leptin, lipids, and glucose tolerance in ovariectomized rats and to compare them to the effects of 17beta-estradiol. DESIGN: Twenty-eight female Sprague-Dawley rats were ovariectomized and fed soy-free chow with the addition of estradiol-3 benzoate (E2B) (10 mg/kg, n=10) or equol (400 mg/kg, n=10). The control group (n=8) received soy-free chow only. Weight and food intake were recorded once weekly. After 6 weeks, intra-abdominal fat was measured using computed tomography, and the intraperitoneal glucose tolerance test was performed. In the seventh week, the animals were killed, blood was collected for plasma, and uteri were removed. RESULTS: Dietary equol significantly increased uterine mass. This effect was, however, 3.5 times lower in magnitude compared to E2B. Similar to E2B, dietary equol decreased weight gain, intra-abdominal fat accumulation, and plasma leptin levels. Equol-treated animals had also lower plasma total cholesterol and triglyceride levels compared to controls. E2B treatment also decreased plasma total cholesterol as well as high-density lipoprotein and low-density lipoprotein cholesterol. In the glucose tolerance test, the area under the curve was significantly smaller in the E2B- and equol treated animals compared to controls. Also, E2B-treated animals had lower fasting plasma insulin levels. CONCLUSIONS: In ovariectomized rats, dietary equol administration attenuates weight gain and shows favorable metabolic effects. However, because of its mild uterotrophic activity, its use in the prevention of postmenopausal weight gain and related metabolic disorders in women with an intact uterus is questionable in terms of safety and warrants further studies. PMID- 17414094 TI - Intracranial complications of cochlear implantation. AB - OBJECTIVE: To describe intracranial complications after cochlear implantation in the pediatric and adult populations. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary referral center. PATIENTS: A chart review of the intracranial complications and their management in 345 patients undergoing cochlear implantation was undertaken. INTERVENTIONS: Variables, including age, sex, implant manufacturer, cause of deafness, intraoperative findings, and postoperative complications, were collected and analyzed. MAIN OUTCOME MEASURE: Presence of intracranial complication of cochlear implantation. RESULTS: There were 134 Nucleus-22 (Cochlear, Englewood, CO) devices, 50 Nucleus-24 devices, 118 Med-El (Durham, NC) devices, and 43 Advanced Bionics Corporation (Sylmar, CA) devices in 151 adults and 194 children. There was a 9.3% overall complication rate, with most (59%) being related to device failure. There were three intracranial complications (<1%), two in elderly individuals and one in a child. Two minor dural defects with cerebrospinal fluid leak at the site of the receiver/stimulator recess in Med-El devices were repaired intraoperatively with temporalis fascia. One elderly patient experienced an acute extensive subdural hematoma after Nucleus-24 implantation, which was treated successfully with immediate evacuation. CONCLUSION: Intracranial complication rates associated with cochlear implantation are low, although potentially very serious. Surgeons should be aware of intracranial complications, especially in older individuals, and take immediate appropriate action. PMID- 17414095 TI - Cystic nephroma and mixed epithelial and stromal tumor of kidney: a detailed clinicopathologic analysis of 34 cases and proposal for renal epithelial and stromal tumor (REST) as a unifying term. AB - Cystic nephroma (CN) and mixed epithelial and stromal tumor (MEST) are rare benign renal neoplasms that have overlapping clinical and morphologic features, including predominance in middle-aged women, variably cystic architecture, eosinophilic cells, and hobnail cells lining the cysts and ovarian-type stroma. The aim of this study was to analyze and compare the histologic features and immunohistochemical profile of these tumors. We studied 34 cases from 5 large academic institutions. Twenty tumors were diagnosed as CNs, 18 in women and 2 in men, their age ranged from 24 to 63 (mean 48; median 50) years. Fourteen tumors were diagnosed as MESTs, all in women, their age ranged from 26 to 84 (mean 52; median 51) years. Histologically, all tumors were well-circumscribed except for one MEST. The stromal/epithelial ratio was approximately 2.3 in MESTs versus 0.3 in CNs; cellular ovarian-type stroma composed 45% of the stroma in MESTs and 12% of the stroma of CNs. Stromal hyalinization was prominent in both. Five MESTs showed stromal luteinization. In the epithelial component, the relative amount of large cysts, medium to small cysts, and phyllodes-type glands was: 65%/25%/10% in CNs versus 25%/40%/35% in MESTs. The epithelial component ranged from flat to cuboidal to hobnail cells in both types of tumors. No significant atypia of either component was seen, although the epithelial cells showed reactive changes. Immunohistochemical stains for estrogen receptors and progesterone receptors showed 62% and 85% positivity in the stromal component of MESTs versus 19% and 40% in CNs. CD10 positivity was seen in 77% of MESTs versus 50% of CNs, calretinin was seen in 69% of MESTs versus 41% of CNs, and inhibin in 42% of MESTs versus 36% of CNs, although the staining was focal. Follow-up in both categories of tumors (mean 3.2 y, median 3 y for CNs and mean 2.5 y, median of 2 y for MESTs) showed no evidence of recurrence or metastases in keeping with their benign nature. This study highlights the remarkable similarity between CN and MEST in sex predilection, age distribution, and morphologic attributes of both the epithelial and stromal components and immunohistochemical profile albeit with variation in individual categories with higher prevalence of stromal to epithelial ratio, prominent ovarian stroma, smaller cysts with phyllodes glands pattern and stromal luteinization being more common in MEST; and large cysts, thin septae and low stromal to epithelial ratio in CN. The presence of ovarian type stroma and mullerian related immunohistochemical markers raises the possibility that these tumors may originate from mullerian remnants misplaced during embryogenesis. On the basis of detailed morphologic analysis of this series of CN and MEST, we propose a unifying term of "renal epithelial and stromal tumor" (REST) to encompass the spectrum of findings observed in these tumors at least until new molecular studies can prove or disprove this challenging hypothesis. PMID- 17414096 TI - Sporadic invasive breast carcinomas with medullary features display a basal-like phenotype: an immunohistochemical and gene amplification study. AB - It is not clear whether invasive breast carcinomas with medullary features (IBCMFs, atypical medullary carcinomas) constitute a specific phenotype of breast cancer that is of biologic significance. Because medullary features are common in BRCA1-associated carcinomas and these tumors frequently show a basal-like phenotype, we examined whether IBCMFs expressed basal/myoepithelial markers and had a basal-like phenotype. We studied the immunohistochemical expression of 15 markers in tissue microarrays containing samples from 35 IBCMFs and 39 grade 3 invasive ductal carcinomas (IDCG3s) of no special type. In addition, we analyzed EGFR, C-MYC, and CCNE gene amplification by fluorescence in situ hybridization, because the expression of these genes is known to be associated with the basal like phenotype. We defined the basal-like phenotype according to the criteria of Nielsen et al as being those tumors that were ER/HER2-negative and cytokeratin (CK) 5/6- and/or epidermal growth factor receptor-positive. IBCMFs were more frequently hormone receptor- and HER2-negative, but had greater expression of proliferation markers and p53. In addition, IBCMFs more frequently expressed basal/myoepithelial markers, such as CK5/6 and P-cadherin. A basal-like phenotype was found in 62.9% of IBCMFs but in only 18.9% of IDCG3s. No differences in gene amplification were found between IBCMFs and IDCG3s, although C-MYC amplification was more common in tumors without a basal-like phenotype. The identification of IBCMF as an independent group of tumors could be of clinical significance, given the high incidence of cases with a basal-like phenotype, which is a group of tumors with different prognosis and chemotherapy response from those of IDCG3s of no special type. PMID- 17414097 TI - Inflammatory myofibroblastic tumor: comparison of clinicopathologic, histologic, and immunohistochemical features including ALK expression in atypical and aggressive cases. AB - Inflammatory myofibroblastic tumor (IMT) is a neoplasm of intermediate biologic potential. In this study, we report a subset of IMTs with histologic atypia and/or clinical aggressiveness that were analyzed for clinicopathologic features, outcome, and immunohistochemical expression of anaplastic lymphoma kinase (ALK) and other markers to identify potential pathologic prognostic features. Fifty nine IMTs with classic morphology (5 cases), atypical histologic features (21 cases), local recurrence (27 cases), and/or metastasis (6 cases) were studied. Immunohistochemistry was performed for ALK1 and other markers (Mib-1, c-Myc, cyclin D1, caspase 3, Bcl-2, Mcl-1, survivin, p27, CD56, p53, MDM-2) using standard techniques. The 59 IMTs had an age at diagnosis ranging from 3 weeks to 74 years (mean 13.2 y, median 11 y, 44% in the first decade). The mean tumor size was 7.8 cm. Sites included the abdomen or pelvis in 64%, lung in 22%, head and neck in 8%, and extremities in 5%. The follow-up ranged from 3 months to 11 years, with a mean of 3.6 years and a median of 3 years. Thirty-three patients had local recurrences, including 13 with multiple local recurrences and 6 patients with both local recurrences and distant metastases. Six patients died of disease, 5 with local recurrences, and 1 with distant metastases. Histologic evolution to a more pleomorphic cellular, spindled, polygonal, or round cell morphologic pattern was observed in 7 cases. Abdominal and pelvic IMTs had a recurrence rate of 85%. Recurrent and metastatic IMTs were larger, with mean diameters of 8.7 and 11 cm, respectively. Cytoplasmic ALK reactivity was seen in 56%. ALK-negative IMTs occurred in older patients (mean age 20.1) years and had greater nuclear pleomorphism, atypia, and atypical mitoses. All 6 metastatic IMTs were ALK-negative. Nuclear expression of p53 was detected in 80% of IMTs overall, but in only 25% of the metastatic subset. There were no significant differences among the subgroups for c-Myc, cyclin D1, MDM-2, Mcl-1, Bcl-2, CD56, p27, caspase 3, or survivin expression. In conclusion, among these 59 IMTs, ALK reactivity was associated with local recurrence, but not distant metastasis, which was confined to ALK-negative lesions. Absent ALK expression was associated with a higher age overall, subtle histologic differences, and death from disease or distant metastases (in a younger subset). Other proliferative, apoptotic, and prognostic markers did not correlate well with morphology or outcome. Thus, ALK reactivity may be a favorable prognostic indicator in IMT and abdominopelvic IMTs recur more frequently. PMID- 17414098 TI - Autoimmune pancreatitis: frequency, IgG4 expression, and clonality of T and B cells. AB - Autoimmune pancreatitis (AIP) is a newly recognized disease. The presence of IgG4 positive plasma cells is thought to be of diagnostic help. In a surgical series of chronic pancreatitis cases, we determined the relative frequency of AIP before and after 1990, analyzed the diagnostic significance of IgG4 expression and examined the presence of oligoclonal T or B-cell populations. The histopathology of 202 surgical specimens of chronic pancreatitis removed between 1975 and 2004 was reviewed and 2 groups were distinguished, 1 of AIP cases and the other of nonautoimmune chronic pancreatitis (non-AIP CP). The intensity of infiltration of pancreatic tissue by IgG4 positive plasma cells and other immune cells was studied immunohistochemically. Finally, T and B-cell clonality was tested by polymerase chain reaction-based analysis. Except for 1 case in 1978, all cases of AIP were observed after 1990. IgG4 positive plasma cells were detected in 72.5% of AIP cases and in 63.1% of non-AIP CP cases. More than 20 cells per high power field were only seen in AIP (sensitivity 43%, specificity 100%). This finding was associated with higher age and grade. Polyclonal T and B-cell populations were found in both AIP and non-AIP CP except for 1 AIP case showing an oligoclonal IgGH-FR3 gene rearrangement. AIP seems to have increased considerably in frequency in the last 2 decades. High density infiltrates of IgG4 positive plasma cells are diagnostic for AIP, but are seen in less than half of the cases. T or B cell oligoclonality could not be established as a feature of AIP. PMID- 17414099 TI - Verruca vulgaris of the vulva in children and adults: a nonvenereal type of vulvar wart. AB - Condyloma acuminata are common lesions of the vulva in adults, and associated with infection by human papillomavirus (HPV) types 6 and 11, which are acquired via sexual contact. The detection of an HPV 6/11 condyloma in the genital tract of a child, therefore, raises the question of sexual abuse. In this study, 29 genital warts in girls less than 5 years of age were examined for nongenital and genital tract HPVs by in situ hybridization. These results were compared with 275 vulvar lesions clinically suspicious for condyloma from adults. Of the 27 HPV related lesions in young girls, 11 (41%) were due to HPV 2 whereas the other 16 (59%) were associated with HPV 6/11 infection. Of the 214/275 (78%) HPV positive vulvar lesions in adults, 6 (3%) were due to HPV 2 whereas 202/214 (94%) contained HPVs 6/11; 1 lesion contained HPV 16 and the 5 other lesions contained HPV 42, 43, or 44. Histologic correlation documented that the vulvar lesions positive for HPV 2 commonly showed the marked hyperkeratosis typical of verruca vulgaris. However, the verrucous pattern was present in lesions HPV 6/11 positive. It is concluded that verruca vulgaris of the vulva, which is likely not transmitted sexually, can occur, albeit rarely, in the genital tract of women and is common in the genital tracts of young girls. This highlights the value of HPV testing in such cases, especially if the histologic changes are consistent with verruca vulgaris. PMID- 17414100 TI - Cholesterol embolization in renal allografts: a clinicopathologic study of 12 cases. AB - Cholesterol embolization (CE) in renal allografts is a rare occurrence, the natural history and prognostic significance of which is poorly characterized. We studied the clinicopathologic features and outcome of the largest known series of CE in renal allografts and combined our cases with those in the literature. We identified renal allograft biopsies with CE from 1997 to September 2004 at University of Pittsburgh Medical Center (UPMC). All pathology material related to such biopsies were examined and correlated with clinical information to determine the most probable CE source. Among 5435 RAB, 19 from 12 cadaveric transplant recipients comprising 7 males and 5 females (median age=63 y) had CE. Donors consisted of 9 males and 2 females (median age=47 y). One donor's age and sex was unknown. The most probable CE source was recipient in 9 cases and donor in 3 cases. Five had acute renal failure without acute cellular rejection and 2 had CE specific failed allografts. Of 19 RAB, the most frequent coexisting diagnosis was chronic allograft nephropathy (63%). The median follow-up time was 661 days. Combining UPMC and non-UPMC cases (n=37) revealed a statistically significant loss of grafts with donor-derived (P value=0.00459) and early CE (P value=0.00938). In renal allografts, CE most often correlated with recipient and donor atherosclerosis. It may present with acute renal failure, but usually not acute graft loss. Graft failure is significantly associated with donor-derived and early CE. Although its prognosis may be poor in the setting of primary nonfunction, prolonged graft survival may be seen. PMID- 17414101 TI - Is stromal microinvasion in primary mucinous ovarian tumors with "mucin granuloma" true invasion? AB - Stromal microinvasion has been recently described in ovarian mucinous borderline tumors (MBTs), leading to proposals for new classifications, including a category of MBTs with stromal microinvasion. This study was conducted to test the validity of this belief. To determine whether stromal microinvasion can be distinguishable from extruded neoplastic epithelium from an adjacent ruptured gland, particularly if accompanied by a mucin granuloma (MG) on hematoxylin and eosin (H&E)-stained sections, we compared the histopathologic features of 138 primary ovarian mucinous tumors, consisting of 81 MBTs, 37 MBTs with stromal microinvasion, 11 intraglandular carcinomas, 2 with microinvasive foci, and 7 mucinous adenocarcinomas with extensive stromal invasion. Immunohistochemical analysis for cytokeratin was performed in 72 cases. Of 77 cases containing MGs, 52% contained "microinvasive foci," consisting of individually scattered epithelial or irregular glandular components in the intervening stroma, as determined by H&E staining. Of the 37 negative cases on H&E, cytokeratin reactivity was detected in 15 additional cases with epithelial/glandular components. Therefore, "stromal microinvasion" can be missed on H&E. All patients with MBT with or without microinvasive foci and localized intraglandular carcinoma with or without microinvasive foci had stage I tumors; none died of tumor-related causes during the follow-up period. Tumor-related deaths were identified only in 5 patients with extensive stromal invasion. This study suggests that some cases of stromal microinvasion in stage I MBTs not accompanied by high grade nuclear atypia represent mucocele-like stromal reaction to ruptured mucinous glands rather than true stromal invasion because isolated tumor cells were exclusively confined to the boundary of MGs and devoid of obvious cellular atypia, and no patient with stromal microinvasion died of tumor-related causes. PMID- 17414102 TI - Immunohistochemical expression of p16 and Ki-67 correlates with degree of anal intraepithelial neoplasia. AB - Anal intraepithelial neoplasia (AIN) is a human papilloma virus related lesion. It has been shown that infection with high-risk human papilloma virus results in up-regulation of p16 and increased cellular proliferation. The objective of this study is to correlate p16 expression and cellular proliferation measured by Ki-67 staining with the degree of dysplasia in the anal canal and to determine the efficacy of these markers in diagnosing high-grade AIN. Seventy-five anal specimens from 55 patients (37 men; 18 women; mean age: 48 y; median: 44 y; range 25 to 96 y) were studied including 35 normal/reactive lesions, 23 low-grade AIN (AIN I and condyloma), and 17 high-grade AIN (AIN II and III). Immunostaining for p16 and Ki-67 was performed. Expression of p16 in AIN correlated with that of Ki 67 (P<0.001). High-grade AIN often demonstrated p16 staining in more than one third of the thickness of the epithelium in a diffuse/continuous fashion. p16 expression in low-grade AIN was often restricted to the lower 1/3 of the epithelium and/or was focal and discontinuous. The expression of both p16 and Ki 67 correlated with the degree of dysplasia (P<0.01). When positive p16 staining was defined as the presence of diffuse/continuous staining in more than one-third of the thickness of epithelium, the sensitivity, specificity, and accuracy of p16 as a marker for diagnosing high-grade AIN were 76%, 86%, and 84%, respectively. When positive Ki-67 staining was defined as the presence of nuclear staining in more than 25% of the cells in more than one-third of the thickness of epithelium, the sensitivity, specificity, and accuracy of Ki-67 as a marker for diagnosing high-grade AIN were 71%, 84%, and 83% respectively. Both p16 and Ki-67 are reliable markers for diagnosing high-grade AIN. PMID- 17414103 TI - Pneumocytic adenomyoepithelioma: a distinctive lung tumor with epithelial, myoepithelial, and pneumocytic differentiation. AB - Pulmonary tumors with epithelial and myoepithelial differentiation are rare, thought to be of bronchial minor salivary gland origin and classified similarly to salivary gland neoplasms. We report a series of a distinctive subtype of pulmonary glandular tumors showing epithelial and myoepithelial differentiation with further pneumocytic specialization. All patients were women, aged 52 to 63 years and presented with single or multiple pulmonary nodules. The tumors were grossly circumscribed, 0.8 to 2.6 cm in greatest dimension, and histologically showed glandular and spindle cell differentiation. Some glands were filled with colloidlike secretion and had an inner, cuboidal epithelial cell layer (pankeratin, epithelial membrane antigen, and thyroid transcription factor-1 positive), surrounded by an outer layer of myoepithelial cells merging with foci of spindled myoepithelial cells (high molecular weight keratin, S100, smooth muscle actin, calponin, caldesmon, and p63 positive). There were also some glands lined by a single layer of plump cells that were positive for surfactant protein A in addition to the other epithelial cell markers. Electron microscopy confirmed pneumocytic features in these cells and the myoepithelial nature of the spindled cells. The surgery in all cases was wedge resection of the masses. The biologic behavior to date has been benign. This is the first reported series of a distinctive lung tumor with epithelial, myoepithelial, and pneumocytic differentiation that differs histologically from all previously recognized pulmonary salivary gland-type and pneumocytic tumors. It is a unique benign appearing neoplasm for which the designation pneumocytic adenomyoepithelioma is suggested. PMID- 17414104 TI - Adenocarcinomas of the distal esophagus and "gastric cardia" are predominantly esophageal carcinomas. AB - BACKGROUND: Adenocarcinoma of the distal esophagus and gastric cardia are defined by the relationship of its epicenter to the gastro-esophageal junction, which is presently defined as the end of the tubular esophagus. We have recently suggested that the true gastro-esophageal junction is best defined by the proximal limit of gastric oxyntic mucosa. AIM: To reclassify adenocarcinomas of this region by the relationship of the tumor to the proximal limit of gastric oxyntic mucosa. METHODS: Seventy-four patients who had esophago-gastrectomy for adenocarcinomas in this region were classified as adenocarcinoma of distal esophagus (38 patients) and gastric cardia (36 patients) by present criteria. The epithelial type at the epicenter and distal edge of these tumors was assessed. RESULTS: The epicenter of the tumor in 64 patients with noncircumferential tumors had squamous (5 cases), cardiac (21 cases), oxynto-cardiac (4 cases), and intestinal (Barrett type) (34 cases) epithelia. None had gastric oxyntic mucosa. Of the 10 patients with circumferential tumors, 7 had cardiac or oxynto-cardiac epithelium at the distal tumor edge. CONCLUSIONS: If the gastro-esophageal junction is defined histologically as the proximal limit of oxyntic mucosa, 71/74 patients would be classified as adenocarcinoma of the distal esophagus. The other 3 patients were questionable as to gastric or esophageal origin. We suggest that this reclassification based on the proposed new definition of the gastro-esophageal junction provides an explanation for the epidemiologic relationship that exists between adenocarcinoma of the "gastric cardia" and gastro-esophageal reflux disease. PMID- 17414105 TI - Desmoplastic small round cell tumor of the kidney in childhood. AB - BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare malignant tumor that generally manifests as abdominal paraserosal masses and affects mainly male adolescents and young adults. When presenting within visceral organs, the diagnosis of DSRCT poses significant difficulties. METHODOLOGY: Four primary renal DSRCT in children diagnosed during a 3-year period are the basis of this report. The medical records and pathologic material were reviewed, including immunohistochemical, ultrastructural, and cytogenetic/molecular studies. RESULTS: The age at presentation was 6 to 8 years, and all children presented with a left renal mass. The tumors measured 3.7 to 13.4 cm and consisted of nests, cords, or sheets of small undifferentiated cells with foci of necrosis and calcification. Desmoplasia was not seen. Tumor cells were immunopositive for vimentin, WT-1 (monoclonal and polyclonal), desmin, cytokeratin, and epithelial membrane antigen. A distinct paranuclear dotlike pattern was observed with vimentin and desmin. Tumor cells possessed rare or focal immunoreactivity for platelet derived growth factor-A and transforming growth factor-beta3, which have been implicated in the pathogenesis of desmoplasia in DSRCT. The EWS-WT1 t(11;22)(p13;q12) translocation was demonstrated in all 4 tumors by fluorescence in situ hybridization and/or reverse transcription-polymerase chain reaction. CONCLUSIONS: DSRCT should be considered in the differential diagnosis of renal tumors composed of small round cells. Undifferentiated morphology and lack of desmoplasia contribute to the difficulty in its recognition. Ancillary studies such as immunohistochemistry may suggest the diagnosis, but cytogenetic and molecular genetic studies are required for confirmation. PMID- 17414106 TI - Glomus tumor of the kidney: a report of 3 cases involving renal parenchyma and review of the literature. AB - Glomus tumor is a rare mesenchymal neoplasm affecting the subcutaneous tissue of the distal extremities in the majority of cases. It only rarely involves visceral organs. We report 3 cases of the glomus tumor family in the kidney, a solid glomus tumor, a glomangioma, and a glomangiomyoma. All 3 tumors involved the renal parenchyma and occurred in 3 men aged 36, 81, and 48 years, respectively. All 3 tumors were well-circumscribed and showed morphology otherwise identical to those seen in soft tissue. All 3 tumors were immunoreactive for actin and negative for desmin and S100 and only 1 tumor expressed CD34 in tumor cells. To date, all 3 tumors have followed a benign course without evidence of recurrence or metastasis. This report expands the spectrum of mesenchymal tumors of the kidney. PMID- 17414107 TI - Ovarian sertoli-leydig cell tumors with pseudoendometrioid tubules (pseudoendometrioid sertoli-leydig cell tumors). AB - The propensity for ovarian endometrioid adenocarcinomas to morphologically mimic Sertoli, Sertoli-Leydig, and granulosa cell tumors, is well known. The converse situation, mimicry of an endometrioid neoplasm by a sex cord-stromal tumor, has not been emphasized. In this report, we describe 9 ovarian Sertoli-Leydig cell tumors (5 well differentiated, 4 of intermediate differentiation) with areas containing hollow, sometimes dilated, tubules which resemble endometrioid glands; we refer to these as pseudoendometrioid tubules. The age of the patients ranged from 14 to 57. The tumors, all of which were unilateral except for one, ranged from 3.5 to 19 cm and were variously described as tan, pale, yellow, or gold. The proportion of the tumor made up of pseudoendometrioid tubules ranged from 10% to >90%. When widespread, their presence sometimes resulted in consideration of a borderline endometrioid adenofibroma or a well-differentiated endometrioid adenocarcinoma. However, all the neoplasms contained typical Sertoli tubules and one or more of the characteristic patterns of Sertoli-Leydig cell tumors as well as Leydig cells, although the latter cells were inconspicuous in some cases. Immunohistochemistry, performed in 4 cases, showed that the pseudoendometrioid tubules, as well as the more typical Sertoli cell elements, were either positive for alpha inhibin (3 of 4 cases) or calretinin (3 of 4 cases) or both, although sometimes focally so. These elements were negative with epithelial membrane antigen and cytokeratin 7. In all 4 cases, the pseudoendometrioid tubules were positive with the broad spectrum cytokeratin AE1/3. This report illustrates the potential for ovarian Sertoli-Leydig cell tumors to contain tubules with a pseudoendometrioid appearance which mimic a borderline or malignant endometrioid neoplasm. The presence of more typical Sertoli cell elements and Leydig cells, an absence of squamous elements, endometriosis or associated adenofibroma, and the characteristic immunophenotype assist in diagnosis. PMID- 17414108 TI - Immunophenotypic characterization and quantification of the epithelial inflammatory infiltrate in eosinophilic esophagitis through stereology: an analysis of the cellular mechanisms of the disease and the immunologic capacity of the esophagus. AB - Eosinophilic esophagitis (EE) is an emerging disease caused by dense infiltration of the esophageal epithelium by eosinophilic leucocytes. It is originated from local hypersensitivity to food or airborne allergens. Although the physiopathologic mechanisms of the illness have not been fully discovered, EE is a loss of immunologic tolerance by the esophagus, meaning that it should be considered as an active immunologic organ. In our study, we investigated the immunologic capacity of the epithelium using immunohistochemistry and stereology, to determine the cellular density of eosinophils, T and B lymphocytes, Langerhans cells, mast cells, and cells manufacturing immunoglobulin E in endoscopic biopsies of patients with EE (taken before and after topical treatment with fluticasone propionate) compared with normal individuals and patients suffering from gastroesophageal reflux disease (GERD). We have observed that the density of eosinophils in EE is 300 times greater than in normal conditions and it is only in this disease where eosinophils show signs of activation and degranulation (positivity to major basic protein immunostaining). The number of T intraepithelial lymphocytes also significantly rose in EE, compared with other entities, where CD8 cells were predominant. However, the human esophagus is deficient in B lymphocytes and we only found intraepithelial plasma cells that excreted immunoglobulin E in EE. Under normal conditions mast cells exist in the thickness of the epithelium that are slightly higher in GERD and multiply in density by 17 in EE. Langerhans cells did not show any significant variation in density under the different tested conditions. After topical treatment with steroids, the density of the different cell components fell to similar levels to GERD. Using our study, we can conclude that the human esophagus may contribute to the development of local immunologic responses as it contains all the necessary cell components. EE represents growth of this esophageal capacity and its pathogeny could respond to mixed cellular and humoral mechanisms. PMID- 17414109 TI - Liver failure and need for liver transplantation in patients with advanced hepatoportal sclerosis. AB - Hepatoportal sclerosis (HPS) is one of the causes of noncirrhotic portal hypertension. In general, hepatic synthetic function is preserved and treatment is aimed at relief of the portal hypertension. In this study, we present the clinical and pathologic features of HPS cases who underwent liver transplantation (LT). LT cases with confirmed gross and microscopic diagnosis of HPS are included. Weight of the explanted liver, presence of thrombi in the main blood vessels, and gross and microscopic characteristics were assessed. Clinical information was gathered from chart review. From 1995 to 2004, 8 LT patients were diagnosed with HPS. Cirrhosis resulting from alcohol (2), autoimmune hepatitis (2), and hepatitis B (1), or cryptogenic cirrhosis (3) was the presumed diagnoses pre-LT. Seven patients presented with bleeding varices and 5 had concomitant ascites. At the time of LT, mean values were: prothrombin time of 15.2 seconds, serum albumin 3.2 g/dL, serum bilirubin 3.5 mg/dL, alkaline phosphatase 140 IU/L, aspartate aminotransferase 39.4 IU/L, and alanine aminotransferase 34.7 IU/L. Explanted livers were shrunken, with weights ranging from 715 to 1199 g (mean 934). Nonocclusive portal vein thrombosis was present in 2 patients. On histologic examination, there was dense portal fibrosis, marked phlebosclerosis, and presence of variable degrees of megasinusoid formation. Four livers also had features of incomplete septal cirrhosis. None showed histologic features of the presumed underlying liver disease. In conclusion, HPS can cause hepatic synthetic dysfunction that may necessitate LT. Small liver volume, significant portal fibrosis, and phlebosclerosis may contribute to hepatic parenchymal loss and subsequent synthetic compromise. PMID- 17414110 TI - Blastomycosis: contributions of morphology to diagnosis: a surgical pathology, cytopathology, and autopsy pathology study. AB - Blastomycosis is caused by the inhalation of spores of the dimorphic fungus, Blastomyces dermatitidis. The reporting of this disease is not required by all states. The diagnosis is established by culture or by identification of broad budding yeast forms in tissue or cytology samples. A retrospective review of blastomycosis was conducted using surgical pathology and cytopathology records of a large community based general hospital, for the years 1982 to 2002; the autopsy records of a university referral center were searched for the years 1992 to 2004. Thirteen surgical/cytology cases were retrieved: 8 localized to the lung (group 1) and 5 with extrapulmonary presentation (group 2). Three of the former were clinically thought to be tumors. Broad-based budding yeast forms with thick cell walls were seen in all but 1 case and identified on conventional routinely stained preparations. Microbiologic culture was positive in 2 lung cases only, in 1 of which it was the sole means of diagnosis. Culture was negative for Blastomyces in the only extrapulmonary case for which a specimen was submitted. One patient in group 2 died, but had a coexistent disseminated gastric adenocarcinoma. No autopsy was performed. Three autopsies of blastomycosis were recovered (group 3). Two patients were on corticosteroids and 1 was diabetic. Premortem diagnoses were established only within a few days of death in 2 patients. Blastomycosis is seldom a fatal disease. Most patients are immune competent; immune compromise favors an aggressive course. Microbiologic culture and conventional morphologic assessment of routine samples have redundant utility in diagnosis. PMID- 17414111 TI - Diffuse pulmonary meningotheliomatosis. AB - Minute pulmonary meningothelial nodules are rare lesions histologically composed of small nests of epithelioid cells located within the interstitium of the lung. These nodules are generally asymptomatic and are usually found incidentally at autopsy or in surgical specimens resected for unrelated causes. The lesions are most often single, although multiple lesions with unilateral involvement of one or even all lobes of the same lung have been described. To our knowledge, cases of meningothelial nodules with disseminated bilateral pulmonary involvement associated with clinical symptoms of restrictive pulmonary disease and radiologic evidence of diffuse reticulonodular pulmonary infiltrates have not been previously documented. We have studied 5 patients presenting with diffuse bilateral pulmonary involvement by numerous minute pulmonary meningothelial nodules. The patients were 4 women and a man aged 54 to 75 years who presented clinically with dyspnea and shortness of breath and the lesions were discovered on open lung biopsies performed for the evaluation of diffuse bilateral interstitial lung infiltrates found on chest x-rays and computed tomography scans. In 3 patients, there was a previous history of malignancy and the radiologic findings were suspected of representing diffuse metastatic disease. Histologically, the lesions were composed of small clusters of epithelioid cells with round to oval nuclei devoid of atypia and surrounded by abundant eosinophilic cytoplasm. Immunohistochemical studies showed positivity of the tumor cells for epithelial membrane antigen and vimentin, and negative staining for cytokeratin, actin, S-100 protein, CD34, chromogranin, and synaptophysin. Electron microscopic examination in 1 case confirmed the ultrastructural features of meningothelial cells, including complex cytoplasmic interdigitations joined by well-developed desmosomes and abundant intracytoplasmic intermediate filaments. The diffuse bilateral involvement of lung parenchyma in the present cases can lead to confusion on clinical and radiologic grounds with a variety of interstitial pulmonary processes, including idiopathic interstitial pneumonia and lymphangitis carcinomatosa. Diffuse pulmonary meningotheliomatosis should be considered in the clinical differential diagnosis of diffuse interstitial pulmonary infiltrates. PMID- 17414112 TI - Myxoid renal tumor with myoepithelial differentiation mimicking a salivary gland pleomorphic adenoma: description of a case. AB - We herein report an unusual case of a low-grade myxoid renal epithelial neoplasm, with peculiar and previously unreported morphologic and immunohistochemical features. The lesion was characterized by noninfiltrative borders, myxoid stroma and elongated tubular and cordlike epithelial structures. These were lined by 2 different epithelial cell types, flat and elongated basal cells and cuboidal to spindle shaped eosinophilic luminal cells, with low-grade nuclear features and a few small nucleoli. The lesion morphologically resembled a pleomorphic adenoma of the salivary gland. The immunohistochemical profile interestingly confirmed the myoepithelial differentiation of the basal epithelial layer, as demonstrated by the coexpression of several myoepithelial markers such as p63, caldesmon, calponin, smooth muscle actin, and S-100, together with epithelial markers such as low and high-molecular weight cytokeratins. The tumor proved benign at follow up. A definitive classification and histogenetic interpretation of this previously unreported tumor type awaits description of further cases showing similar features which, perhaps, as it may happen, went so far unnoticed. PMID- 17414113 TI - CDKN2A (INK4A-ARF) mutation analysis to distinguish cutaneous melanoma metastasis from a second primary melanoma. AB - The histologic differential diagnosis between a second primary cutaneous melanoma and cutaneous melanoma metastasis in a patient with a previous history of melanoma can be very difficult. This case report describes the first application of CDKN2A mutation analysis for discriminating a cutaneous melanoma metastasis from a new primary melanoma. In 2005, we received a skin excision of the right arm of a 38-year-old female patient for second opinion. Histologically, we considered the lesion to be a melanoma. The patient had a history of superficial spreading melanoma in the right subclavicular region, with a Breslow thickness of 1.1 mm, in 1998. The morphology showed resemblance to the present melanoma on the right arm, but the differential diagnosis between metastasis or second primary melanoma could not be made with certainty based on histology alone. We decided to perform TP53 and CDKN2A mutation analysis on both tumors. Molecular analysis revealed that both the melanoma of 1998 and of 2005 contained an identical CDKN2A mutation (a deletion in exon 1alpha, c.95_112del (p.Leu32_Leu37del)), which was absent in normal control tissue of the patient, thereby excluding a germline mutation. TP53 mutations were absent in both tumors and in normal skin. Based on these molecular findings the present melanoma on the right arm was diagnosed as a metastasis. Seven months later the patient died of widespread metastatic disease confirming the metastatic nature of the lesion. This case illustrates that molecular analysis can contribute to the sometimes-difficult differentiation between a second primary melanoma and a melanoma metastasis. PMID- 17414115 TI - Pseudosarcomatous myofibroblastic proliferations of the bladder: a clinicopathologic study of 42 cases. PMID- 17414116 TI - Chronic sclerosing dacryoadenitis: part of the spectrum of IgG4-related Sclerosing disease? PMID- 17414117 TI - Familial and sporadic pancreatic carcinoma, epidemiologic concordance. PMID- 17414118 TI - Gastric epithelial siderosis in acute lymphoblastic leukemia. PMID- 17414119 TI - Fauniform changes in cytopathology. PMID- 17414120 TI - The 2006 Fred Waldorf Stewart Award recipient Juan Rosai, MD. PMID- 17414122 TI - Radiological imaging in pneumonia: recent innovations. AB - PURPOSE OF REVIEW: Pneumonia is one of the major infectious diseases responsible for significant morbidity and mortality throughout the world. Radiological imaging plays a prominent role in the evaluation and treatment of patients with pneumonia. This paper reviews recent innovations in the radiologic diagnosis and management of suspected pulmonary infections. RECENT FINDINGS: Chest radiography is the most commonly used imaging tool in pneumonias because of availability and an excellent cost-benefit ratio. Computed tomography is mandatory in unresolved cases or when complications of pneumonia are suspected. A specific radiologic pattern can suggest a diagnosis in many cases. Bacterial pneumonias are classified into four main groups: community-acquired, aspiration, healthcare associated and hospital-acquired pneumonia. The radiographic patterns of community-acquired pneumonia may be variable and are often related to the causative agent. Aspiration pneumonia involves the lower lobes with bilateral multicentric opacities. The radiographic patterns of healthcare-associated and hospital-acquired pneumonia are variable, most commonly showing diffuse multifocal involvement and pleural effusion. SUMMARY: Combination of pattern recognition with knowledge of the clinical setting is the best approach to the radiologic interpretation of pneumonia. Radiological imaging will narrow the differential diagnosis of direct additional diagnostic measures and serve as an ideal tool for follow-up examinations. PMID- 17414123 TI - Community acquired pneumonia in the tropics. AB - PURPOSE OF REVIEW: Due to the exponential growth of international exchange, millions of travelers are exposed to respiratory pathogens in the tropics and may return ill. Community-acquired pneumonia is one of the more prevalent infections. RECENT FINDINGS: The acquisition of infections in the tropics, including community-acquired pneumonias, has been described for several centuries. During recent decades some microorganisms have been disclosed as causative of the disease (Legionella pneumophila in 1976 and hantavirus in 1990); other microorganisms are real new pathogens that were not previously demonstrated to have a pathogenic role in humans (e.g. severe acute respiratory syndrome coronavirus producing an outbreak in 2003 and H5N1 avian influenza virus producing an increasing number of human cases over the last few years). SUMMARY: A number of microorganisms may produce pneumonia in people who live or have traveled to tropical zones. History, including geography and epidemiology, physical exam and complementary workout are precious tools for the diagnosis, therapy and prevention. Exposure to microorganisms in tropical areas may show different patterns. A high index of suspicion, detailed investigation of travel, exposure history of the patient, and a basic understanding of the incubation periods and distribution of the various potential pathogens are imperative for the diagnosis. PMID- 17414124 TI - Community-acquired pneumonia: what is relevant and what is not? AB - PURPOSE OF REVIEW: Community-acquired pneumonia is associated with significant morbidity and mortality and is the most common cause of death from infectious diseases in North America. The purpose of this review is to highlight recent advances in epidemiology, risk factors, severity criteria and antibiotic therapeutic regimens used for community-acquired pneumonia management. RECENT FINDINGS: All guidelines recommend early and appropriate empiric therapy directed against common typical organisms, such as Streptococcus pneumoniae, and other atypical organisms, but clinicians should be aware of newer emerging pathogens such as community-acquired methicillin-resistant Staphylococcus aureus and Gram negative pathogens. SUMMARY: The optimum outcome in community-acquired pneumonia can be achieved by careful risk stratification using prediction rules together with appropriate antibiotic regimens. The mainstay of community-acquired pneumonia prevention is influenza and pneumococcal immunization. Promotion of smoking cessation will also help curtail the incidence of pneumococcal disease. PMID- 17414125 TI - Diagnostic strategies for nosocomial pneumonia. AB - PURPOSE OF REVIEW: This review describes advances in clinical and microbiological modalities for diagnosis of nosocomial pneumonia and the role of biological markers. RECENT FINDINGS: Serial assessments with the clinical pulmonary infection score identifies nonsurvivors and allows discontinuation of antibiotics when there is low suspicion of pneumonia. Studies evaluating its clinical utility show mixed results. A meta-analysis revealed that an invasive approach does not affect mortality but reduces costs, antibiotic exposure, and multidrug resistance. In contrast to these findings, a recent trial comparing nonquantitative endotracheal aspirate and quantitative bronchoalveolar lavage cultures showed similar clinical outcomes and antibiotic utilization. The role of quantitative endotracheal aspirate for diagnosis of pneumonia not related to mechanical ventilation was recently evaluated. Procalcitonin and soluble triggering receptor expressed on myeloid cells-1 aid in diagnosis, identify sepsis related to ventilator-associated pneumonia and patients with worst outcomes. SUMMARY: The diagnostic modality chosen depends on availability, personnel experience, and the patient's clinical status. Recent guidelines support the use of quantitative cultures in an integrated clinical and microbiological algorithm. The decision to adjust antibiotics involves clinical reassessment and interpretation of culture results. Biological markers have a potential role as screening and prognostic tools. PMID- 17414126 TI - Aspiration pneumonias: under-diagnosed and under-treated. AB - PURPOSE OF REVIEW: To examine recent advancements of the epidemiology, risk factors, bacteriology, diagnostic measures, and treatment for aspiration pneumonia. RECENT FINDINGS: In a large prospective study, 10.3% of community acquired pneumonia requiring hospitalization was associated with aspiration pneumonia, as opposed to 30% when patients were admitted from long-term facilities. A murine stroke model showed increased apoptosis of cell-mediated immune cells due to overactivation of the sympathetic nervous system. Subsequent nasal inoculum of low-dose Streptococcus pneumoniae caused severe pneumonia. Another murine study showed intratracheal administration of hydrochloric acid primed lungs more susceptible to infection with Klebsiella pneumoniae. Tracheal pH monitoring demonstrated acidification of trachea in 28% of patients with acute stroke deemed to be safe for oral feeding. Various antimicrobial agents used to treat aspiration pneumonia have comparable success rates. However, clindamycin was the only agent that did not propagate emergence of methicillin-resistant Staphylococcus aureus. SUMMARY: Aspiration pneumonia is prevalent in community acquired pneumonia. Further studies are needed to elucidate the effects of sympathetic nervous system and aspiration pneumonitis as primers for lung infection. Predicting aspiration risk is difficult and newer diagnostic tools are necessary. Antimicrobials used in aspiration pneumonia have similar success rates although the development of resistant organisms must be monitored. PMID- 17414127 TI - Emerging fungal pathogens in pulmonary disease. AB - PURPOSE OF REVIEW: Invasive fungal infections of the lung have been increasing due to the increase of the population most at risk. This review aims to describe some of the emerging fungal pathogens and their complex management. RECENT FINDINGS: With the increase in immunosuppressed populations, physicians are increasingly encountering uncommon fungal pathogens that historically have been difficult to identify and treat. Many of these fungal infections present with similar clinical features and often show similar histopathological changes. Treatment options are more complex because of an increasing number of antifungals that have become available for clinical use. The correct usage of these antifungals in addressing emerging fungal infections is unclear, however. Drawing from in-vitro and in-vivo susceptibility testing and case reports, some deductions may be made for the best empirical treatment of these deadly diseases. In general, the newer triazoles (voriconazole and posiconazole) and the use of combination therapy have shown promise. SUMMARY: Invasive fungal infections are on the increase and contribute significantly to overall mortality, particularly among transplant recipients. With the lack of well designed controlled clinical studies, physicians will need to draw from previously described cases and in vitro susceptibility testing to optimize therapy. PMID- 17414128 TI - Tuberculosis and pregnancy. AB - PURPOSE OF REVIEW: To provide a summary of the diagnostic and therapeutic challenges, including risks and benefits of treatment, of tuberculosis and latent Mycobacterium tuberculosis infection during pregnancy. RECENT FINDINGS: Recent developments in diagnostic options have added to the armamentarium of tests available to diagnose latent Mycobacterium tuberculosis infection. Increasing evidence supports the potential for successful treatment of multidrug-resistant tuberculosis during pregnancy with good maternal and neonatal outcomes. The impact of genital tuberculosis on the outcome of assisted in-vitro fertilization techniques is noted. SUMMARY: The diagnostic approach for the evaluation of tuberculosis or latent Mycobacterium tuberculosis infection is unchanged by pregnancy, and includes clinical suspicion of disease, tuberculin skin testing or interferon-gamma-based assay, chest radiography with appropriate shielding when indicated, and acid-fast bacillus stain and culture of clinical material. For patients with active tuberculosis, therapy should be initiated as soon as the diagnosis is established. Initiation of treatment for latent infection during pregnancy should be considered based on the risk for progression to active disease. PMID- 17414129 TI - The clinical management of drug-resistant tuberculosis. AB - PURPOSE OF REVIEW: Drug-resistant tuberculosis is a growing problem, with almost half a million cases worldwide. In spite of the difficulty in its management, drug-resistant tuberculosis can be successfully treated, even in poor settings. RECENT FINDINGS: This article will review key findings in the areas of epidemiology, diagnosis and management of drug-resistant tuberculosis, including new antituberculous drugs. The issue of extensively drug-resistant tuberculosis will also be reviewed and discussed. Finally, novel approaches to the management of drug-resistant tuberculosis in populations with HIV, as well as in pediatric populations, among pregnant women, and among patients requiring surgical therapy, will be reviewed. SUMMARY: New advances in the diagnosis and management of drug resistant tuberculosis allow for excellent clinical outcomes to be achieved, even in difficult-to-treat populations. This is possible with timely diagnosis of disease and rapid initiation of appropriate therapy in supported settings. PMID- 17414130 TI - How to diagnose and manage common parasitic pneumonias. AB - PURPOSE OF REVIEW: Parasitic pneumonia is being increasingly reported from many parts of the world due to globalization and travel across the continents. This review outlines the recent developments in the diagnosis and management of parasitic pneumonias. RECENT FINDINGS: A polymerase chain reaction that can differentiate pathogenic Entamoeba histolytica from nonpathogenic species has been reported. It has been observed that pulmonary infection with Leishmania donovani can occur in immunodeficient and lung transplant patients. Acute respiratory distress syndrome, seen in severe falciparum malaria, has also been observed in vivax malaria. A study has demonstrated the return of chloroquine sensitive falciparum malaria several years after chloroquine treatment was discontinued. Pulmonary hypertension has been reported in Schistosoma hematobium, S. mansoni and S. japonicum infections. Strongyloides hyperinfection and disseminated disease are frequently reported in immunocompromised individuals. Parenteral ivermectin is found to be useful in the treatment of disseminated strongyloidiasis. A chronic mild interstitial lung disease has been found to persist in tropical pulmonary eosinophilia despite treatment. Studies are in progress to develop vaccines against amoebiasis, malaria and hookworm infections. SUMMARY: Parasitic pneumonia can sometimes be life threatening. If proper diagnosis is made early, the pneumonia can be treated successfully with currently available drugs. PMID- 17414131 TI - Bronchial lavage in tropical pneumonias. AB - PURPOSE OF REVIEW: In developing countries, where resources are scarce, it is paramount that the clinician be familiar with the most cost-effective diagnostic and therapeutic modalities available. The fiberoptic bronchoscope is an expensive piece of equipment, but can be an economical diagnostic tool in tropical pneumonias. The role of bronchoalveolar lavage is discussed in this review. RECENT ADVANCES: Since its introduction in the 1970s, particularly in developing countries, bronchoalveolar lavage has become an important tool for diagnosis, assessing the activity and monitoring a large number of pulmonary diseases. The importance of bronchoalveolar lavage is becoming even more apparent since the emergence of the HIV/AIDS epidemic, which has changed the face of many pulmonary diseases, especially tropical ones. SUMMARY: Bronchoalveolar lavage is useful in developed and developing countries alike, for assessing many bacterial, fungal, and parasitic pulmonary diseases. PMID- 17414133 TI - Bibliography. Current world literature. Infectious diseases. PMID- 17414132 TI - Leptospiral pneumonias. AB - PURPOSE OF REVIEW: Pulmonary involvement in leptospirosis has been reported to be increasing in the last few years and is emerging as a serious life threat and the main cause of death due to leptospirosis in some countries. In this review, we present the main clinical and pathological manifestations of pulmonary involvement in leptospirosis, and recent data on the pathophysiology of lung damage and therapeutic implications. RECENT FINDINGS: Although previous reports have emphasized the increasing incidence of pulmonary manifestations in patients with leptospirosis worldwide, pulmonary involvement in leptospirosis is still under-recognized. Experimental models have been used to address new aspects of the pathogenesis of the disease, including determination of novel outer membrane proteins, characterization of dysregulation of sodium transporters of alveolar epithelial cells, and provision of new data on the role of innate immunity in the development of severe disease, thereby making major progress in understanding the mechanisms underlying lung injury. SUMMARY: Pulmonary hemorrhage represents the main cause of death in severe forms of leptospirosis. Although the mechanism of pulmonary impairment is still poorly understood, recent experimental studies have brought new insights to the pathogenesis of lung injury and provide new perspectives on treatment of critically ill patients. PMID- 17414134 TI - Reversal of liver cirrhosis: a desirable clinical outcome and its pathogenic background. AB - Cirrhosis is the final stage of chronic liver damage of various etiologies. It used to be considered an irreversible lesion, but enormous advances in our understanding of hepatic cellular and molecular biology in the past 2 decades have challenged this view. There is now substantial evidence that cirrhosis can be a reversible process. This concept is supported by an increasing number of clinical reports showing the disappearance of cirrhotic lesions from liver biopsies taken from patients cured of their liver disease. The reversal of cirrhosis usually occurs in patients with short-lived liver disease, after the successful treatment of the underlying liver damage. Recently, however, we observed the spontaneous reversal of cirrhosis after the loss of hepatitis B viremia in 2 men, 21 and 28 years old, who had developed cirrhosis as young children. Several questions and controversial issues concerning the definition of advanced cirrhosis, the limitations of liver biopsy (eg, sampling, interpretation error), and the applicability of noninvasive methods to the assessment of fibrosis, are being addressed. Future prospects include the possibility of antifibrotic therapy to prevent fibrosis or favor its degradation. PMID- 17414135 TI - Fecal calprotectin in very low birth weight infants. AB - OBJECTIVES: To measure concentrations of fecal calprotectin (f-calprotectin) in infants with very low birth weight (VLBW; <1500 g) longitudinally and to describe changes in f-calprotectin in infants who develop severe abdominal disease. PATIENTS AND METHODS: The study included 59 VLBW infants. Seven patients (disease group) developed severe abdominal disease defined as necrotizing enterocolitis (NEC) or a condition leading to laparotomy. The remainder (n = 52) were considered reference infants and had a mean (+/-SD) gestational age of 27.2 +/- 2.6 weeks and a birth weight of 939 +/- 273 g. F-calprotectin was analyzed in meconium and weekly during postnatal weeks 1 to 8. In disease cases, more frequent samples were analyzed around the time of abdominal disease diagnosis. RESULTS: In reference infants the median (range) f-calprotectin level in meconium was 332 (12-9386) microg/g and correlated negatively to Apgar score. F calprotectin in postmeconium samples was 253 (9-1867) microg/g and correlated positively to delivery by cesarean section, postnatal age, and volume of enteral feeds, and negatively to treatment with antibiotics and corticosteroids. In reference infants no postmeconium sample had f-calprotectin levels >2000 microg/g. In disease cases f-calprotectin was increased to >2000 microg/g in 3 cases of NEC and 1 case of covered perforation with microscopic bowel inflammation. In 1 case of NEC without microscopic bowel inflammation and 2 cases of focal intestinal perforation, f-calprotectin levels never exceeded 2000 microg/g. CONCLUSIONS: F-calprotectin concentrations in VLBW infants are similar to previously reported levels in healthy term and moderately preterm infants. An f-calprotectin level >2000 microg/g is a useful but not an early marker of NEC and other severe intestinal inflammatory conditions in VLBW infants. PMID- 17414136 TI - Fecal lactoferrin is a sensitive and specific marker of disease activity in children and young adults with inflammatory bowel disease. AB - BACKGROUND AND AIMS: Fecal lactoferrin (FLA) is a neutrophil-derived surrogate marker of intestinal inflammation that is elevated in patients with inflammatory bowel disease. However, the correlation between FLA levels and serological markers of disease activity has not been previously reported, to our knowledge. In the present study we evaluated the ability of FLA levels to reflect disease activity in pediatric patients with inflammatory bowel disease. We further assessed the relationship between FLA levels and customary laboratory and clinical measures of inflammation. PATIENTS AND METHODS: Fecal specimens were collected from 148 consecutive pediatric patients (79 with Crohn disease, 62 with ulcerative colitis, and 7 with irritable bowel syndrome) and 22 healthy control individuals. Lactoferrin was measured by enzyme-linked immunosorbent assay (IBD SCAN, TECHLAB, Inc). Disease activity was assessed at the time of sample provision by laboratory measures (including erythrocyte sedimentation rate [ESR] and albumin) and previously validated disease activity indices (Pediatric Crohn Disease Activity Index, Kozarek, Harvey Bradshaw Activity Index). RESULTS: Lactoferrin levels were significantly higher in patients with ulcerative colitis (1880 +/- 565 microg/mL) (mean +/- SE) or Crohn disease (1701 +/- 382 microg/mL) than in healthy control individuals under 21 years of age (1.17 +/- 0.47 microg/mL, P < 0.001). Lactoferrin levels correlated significantly with ESR, hematocrit, albumin, and platelet count (P < 0.001). Receiver operating characteristic curve analysis revealed that FLA levels were comparable to ESR in detecting patients with clinically active disease (P < 0.001). Patients who experienced a clinical flare within 2 months of specimen collection displayed higher lactoferrin levels (845 +/- 452 microg/mL) than did those who remained in clinical remission (190 +/- 90 microg/mL, P = 0.003). CONCLUSIONS: Data presented here demonstrate that FLA is a sensitive and specific biochemical marker of inflammation for use in the diagnosis and interval assessment of pediatric patients with IBD, and its level correlates well with both clinical disease activity indices and ESR. Elevated levels of FLA may also identify patients at greater risk for the development of subsequent clinical flares. PMID- 17414137 TI - Long-term treatment with autologous red blood cells loaded with dexamethasone 21 phosphate in pediatric patients affected by steroid-dependent Crohn disease. AB - BACKGROUND: Inflammatory bowel disease (IBD) present in childhood in 15% to 25% of cases. The aim of therapy in children is not only to guarantee normal growth but also to prevent relapse and to maintain remission. Steroids are effective to induce remission; however, resistance, dependency, and irreversible side effects can develop. The aim of this study was to determine whether treatment with repeated infusions of autologous red blood cells (RBCs) loaded with dexamethasone 21-phosphate (Dex 21-P) is safe and allows maintenance of long-term remission in children with steroid-dependent Crohn disease (CD). PATIENTS AND METHODS: Eighteen consecutive pediatric patients who met the inclusion criteria were admitted to the study. Infusions of autologous RBCs loaded with Dex 21-P were performed every 4 weeks; the mean duration of treatment was 24 months. At the beginning of treatment and after 6, 12, and 24 months, we performed clinical evaluation according to the Pediatric Crohn Disease Activity Index (pCDAI). Assessment of body mass in dexamethasone and bone mineral density by means of computerized bone mineralometry-dual energy x-ray absorptiometry, endoscopic evaluation, and hematic morning cortisol determination were also performed. RESULTS: During treatment, the mean pCDAI significantly decreased (P < 0.05); 78% of patients discontinued steroids. Determination of morning cortisol showed suppression only on the first day after infusion, followed by normalization of values. Endoscopic findings showed remission in 44% of patients. None of the patients experienced serious side effects. CONCLUSIONS: These data suggest that repeated infusions of RBCs loaded with Dex 21-P can be safe and useful to maintain long-term remission in pediatric patients with moderately active CD. PMID- 17414138 TI - Role of methotrexate in the management of Crohn disease. AB - BACKGROUND: The majority of children with Crohn disease (CD) are likely to need some form of immunomodulatory therapy to maintain remission and to avoid long term corticosteroid usage. Although thiopurine agents are commonly used, some children are unresponsive or intolerant to these drugs. Biological agents like infliximab are being increasingly used in these circumstances, but long-term safety has yet to be established. Methotrexate has been shown to induce and maintain remission in CD in many adult studies, but pediatric data are limited. The present report describes our experience of using methotrexate in CD in children. PATIENTS AND METHODS: All children with CD treated with methotrexate were identified by the departmental database. Case records were reviewed for site of disease, Pediatric Crohn Disease Activity Index, medications, time to achieve remission, duration of remission, and complications. RESULTS: A total of 10 children received methotrexate, 7 of whom were female and 3 of whom were male. All of the children had colonic involvement and had active disease despite previous standard medical treatments. Seven children exhibited remission with methotrexate treatment. Median time to achieve remission was 12 weeks and median duration of remission was 21 months to the point of assessment. One child had transient increase of alanine aminotransferase levels and another developed neutropenia, which remitted with dose reduction. None needed discontinuation of methotrexate treatment. CONCLUSIONS: Methotrexate is effective and well tolerated in children with CD. It should be considered in those patients who do not experience a remission with standard medications because it may avoid the use of biological agents and their potential uncertain long-term side effects. PMID- 17414139 TI - Failure to respond to hepatitis B vaccine in children with celiac disease. AB - OBJECTIVES: To determine whether children with celiac disease (CD) fail to show a response to hepatitis B virus (HBV) vaccine more frequently than children without CD. PATIENTS AND METHODS: This was a prospective study that compared the response to HBV, tetanus, rubella, and Haemophilus influenzae type b (Hib) vaccines between children with CD and age- and sex-matched control subjects. RESULTS: The study population included 26 patients with CD and 18 age- and sex-matched controls. All had received the full complement of childhood vaccinations. A significantly higher proportion of subjects in the CD group (14 of 26) failed to respond to HBV vaccine compared with controls (2 of 18; 53.9% vs 11.1%; P < 0.05). Patients with CD were 8.33 times more likely to test negative for hepatitis B surface antigen than control subjects (95% CI, 1.5-46.5). By contrast, all of the subjects in both groups tested positive for rubella antibodies; only 1 subject in the CD group tested negative for tetanus antibody versus none in the control group (3.9% vs 0%; P = 1.0). The percentage of subjects who tested negative for Hib antibodies was similar in the 2 groups (CD, 33.3%; control, 44.4%; P = 0.53). CONCLUSIONS: More than 50% of children with CD do not show a response to standard vaccination regimens for HBV. Given the large number of children with CD throughout the world, this observation suggests that there is a large HBV-susceptible population despite widespread vaccination. Current immunization strategies may need to be reassessed to protect this population and achieve the goal of universal protection. PMID- 17414140 TI - Prospective controlled long-term follow-up for functional outcome after anoplasty in boys with perineal fistula. AB - BACKGROUND: Functional outcomes after treatment of low anorectal anomalies remain controversial. To address this, we conducted a controlled, prospective single center long-term follow-up study of functional outcome in boys treated for perineal fistula with anoplasty. PATIENTS AND METHODS: Twenty-four consecutive boys treated exclusively for perineal fistula from 1992 to 2001 underwent prospective follow-up after completion of toilet training. All of the patients had identical surgical treatment. Functional outcome was assessed according to a previously validated bowel function score (7 questions, maximum score of 20). Daytime and nighttime wetting, as well as the age at toilet training, were also recorded. Thirty-seven age-matched boys served as healthy controls. An independent nurse specialist interviewed caregivers. RESULTS: Age of the patients (9.1 +/- 2.6 years) and controls (9.0 +/- 2.9 years) were similar. Seven patients (32%) had bowel function scores below the 10th percentile (17.2) of the controls, which was chosen as a lower limit of normality. The overall mean bowel function score was significantly lower (P < 0.01) among the patients (17.9 +/- 1.9) compared with the controls (19.1 +/- 1.2). The difference was attributed mainly to significantly lower scores (0-3) among patients in questions assessing constipation (2.4 +/- 0.8 vs 2.9 +/- 0.3; P < 0.05) and soiling (2.5 +/- 0.5 vs 2.8 +/- 0.4; P = 0.05). Constipation and soiling were significantly more common (P < 0.05) among patients (41% and 55%, respectively) compared with controls (8% and 24%, respectively). CONCLUSIONS: Overall long-term bowel function is impaired in one third of boys with perineal fistula. The main reasons for impaired functional outcome are constipation and soiling, which affect as many as half the patients. PMID- 17414141 TI - Appropriateness of upper gastrointestinal endoscopy in children: a retrospective study. AB - BACKGROUND: Upper gastrointestinal endoscopy (UGIE) is appropriate in many situations in adults and children. Recommendations for UGIE use in children were published recently by the French-language Paediatric Hepatology, Gastroenterology, and Nutrition Group (GFHGNP). PATIENTS AND METHODS: We retrospectively reviewed the 293 UGIE procedures undertaken in 251 children between January 1, 2001 and June 30, 2003 by 2 senior endoscopists. The UGIE procedures were categorized as appropriate or inappropriate based on GFHGNP recommendations, and diagnostic efficiency was compared in the 2 groups with the chi2 test followed by multivariate logistic regression analysis. RESULTS: Of the 293 UGIE procedures, 52 (17.7%) were considered inappropriate. Diagnostic efficiency was 51% in the appropriate group versus 17.3% in the inappropriate group (odds ratio, 4.2; 95% CI, 2-8.7; P < 10(-3)). The proportion of appropriate UGIE procedures was higher among inpatients than outpatients (odds ratio, 2.51; 95% CI, 1.24-5.08; P = 0.01). Inappropriate reasons for performing UGIE included isolated failure to thrive and follow-up after neonatal esophagogastroduodenitis. Nine inappropriate UGIE procedures contributed useful information: ulcerative esophagitis in 1 patient, hemorrhagic esophagitis in 4 patients, duodenitis in 1 patient, and malabsorption in 3 patients caused in 1 case by cow's milk allergy and in 2 cases to fully documented celiac disease. CONCLUSIONS: UGIE was usually performed appropriately in our pediatric hospital. Inappropriate UGIE procedures were more common in outpatients than in admitted patients. Awareness of the recommendations for appropriate UGIE use needs to be improved among office-based and hospital-based physicians. PMID- 17414142 TI - Metabolic and nutritional profile of obese adolescents with nonalcoholic fatty liver disease. AB - BACKGROUND: The incidence of nonalcoholic fatty liver disease (NAFLD) is increasing due to its prevalence in obesity, diabetes, and insulin-resistance syndrome. The best treatment protocol for NAFLD has not been determined. However, there is evidence that exercise and nutritional intervention can improve and prevent it. The aim of the present study was to evaluate the dietary and metabolic profiles of obese adolescents with NAFLD who participated in a multidisciplinary program. PATIENTS AND METHODS: We studied 43 adolescents ages 15 to 19 years (17.18 +/- 1.66 years) with a body mass index (BMI) > or = 30, consisting of 30 patients without NAFLD (BMI = 35.80 +/- 3.44 kg/m2) and 13 with NAFLD (BMI = 33.47 +/- 2.34 kg/m2). The NAFLD diagnosis was determined by ultrasonography. Blood samples were collected to analyze glycemia, hepatic aminotransferase levels, and lipid profiles. Insulin resistance was measured by homeostasis model assessment insulin-resistance index (HOMA-IR). The analyses of baseline and postintervention food intake were made by a 3-day inquiry. RESULTS: At baseline conditions, the patients with NAFLD showed significant differences in body mass, BMI, and visceral and subcutaneous fat. Glucose and visceral and subcutaneous fat presented a significant reduction after treatment in patients with NAFLD. Analyzing the food intake, at baseline we observed a positive correlation between the visceral obesity and lipid consumption only in patients with NAFLD. We also observed significant decrease in energy and cholesterol consumption in patients with NAFLD after the multidisciplinary therapy. CONCLUSIONS: The intervention promoted a decrease in the prevalence of NAFLD, a significant decrease in visceral obesity, and improved HOMA-IR, glycemia, and serum lipid levels that are risk factors for NAFLD. In summary, the multidisciplinary program is essential in the treatment and prevention of NAFLD. PMID- 17414143 TI - Prenatal molecular diagnosis of inherited cholestatic diseases. AB - OBJECTIVES: Progressive familial intrahepatic cholestasis (PFIC) and to a lesser extent, Alagille syndrome, often lead to end-stage liver disease during childhood. We report our experience of DNA-based prenatal diagnosis of PFIC1-3 and Alagille syndrome. PATIENTS AND METHODS: Four molecular antenatal diagnoses were performed in 3 PFIC families and 17 in 11 Alagille syndrome families. DNA was isolated from chorionic villus or cultured amniocyte samples from women, without pregnancy complications. RESULTS: All four foetuses with a family history of PFIC1, 2, or 3 were heterozygous for an ATP8B1, ABCB11, or ABCB4 mutation and pregnancies were continued. Three of the infants were healthy after birth, and 1 premature infant, who had an ABCB4 mutation, experienced transient neonatal cholestasis. Among the families with a history of de novo JAG1 mutation, none of the foetuses was mutated, versus 40% of those with a history of familial mutation. Of 4 pregnant women with a JAG1-mutated foetus, 3 cut short their pregnancy and 1 gave birth to a child with overt Alagille syndrome. CONCLUSIONS: Molecular antenatal diagnosis of PFIC1-3 and Alagille syndrome is reliable because clinical outcome after birth corresponded to molecular foetal data. PMID- 17414144 TI - Early central catheter infections may contribute to hepatic fibrosis in children receiving long-term parenteral nutrition. AB - BACKGROUND: Bacterial infections in infants constitute a risk factor for parenteral nutrition (PN)-related cholestasis. The possible role of infections in the development of liver fibrosis, the most severe long-term complication, has yet to be documented. This study retrospectively compares the incidence of sepsis in children with and without severe liver fibrosis. PATIENTS AND METHODS: Medical reports of 30 children in prolonged PN programs between March 1985 and March 2000 were reviewed. Starting at birth, the mean PN duration was 65 months (range, 8 150 months). According to the results of liver biopsy (LB), patients were split into 2 groups: group A (n = 16) with severe liver fibrosis (ie, septal fibrosis involving >50% of portal fields or cirrhosis) and group B (n = 14) with normal hepatic architecture or mild fibrosis (<50% of portal fields). RESULTS: Duration of PN at the time of LB was shorter in group A (30.5 months; range, 8-96 months) than in group B (105 months; range, 37-150 months; P < 0.001). In group A the incidence of sepsis was significantly higher than in group B (3.2 +/- 0.3/year vs 1.5 +/- 0.2/year) and the first infection occurred earlier (group A, 1 month [range, 1-2 months]; group B, 4 months [range, 1-19 months]). By contrast, both groups were similar in terms of pregnancy duration, birth weight, age of PN onset, underlying diseases, mode of PN delivery, and number of cholestasis episodes. CONCLUSIONS: Incidence and early onset of infections may contribute to the development of liver fibrosis in cases of long-term PN. New strategies are required in prevention and treatment of infections in children receiving PN. PMID- 17414145 TI - Decreased human milk concentration of epidermal growth factor after preterm delivery of intrauterine growth-restricted newborns. AB - BACKGROUND AND AIM: The aim of the study was to compare epidermal growth factor (EGF) concentration in 81 colostrum samples collected from mothers of newborns in the following growth categories: preterm appropriate for gestational age (AGA), preterm small for gestational age (SGA), and full term (FT). RESULTS: Significantly higher concentrations of EGF were found in the colostrum of mothers who delivered premature AGA infants at less than 32 weeks of gestation compared with mothers who delivered premature SGA babies at the same gestational age. CONCLUSIONS: We concluded that the maternal compensatory mechanism accelerating the development of immature breast-fed infants may be disturbed when gestation is complicated by intrauterine growth retardation. PMID- 17414146 TI - Effect of iron supplementation on haemoglobin response in children: systematic review of randomised controlled trials. AB - OBJECTIVE: To evaluate the effect of iron supplementation on haemoglobin (Hb) in children through a systematic review of randomised controlled trials. MATERIALS AND METHODS: Electronic databases, personal files, hand search of reviews, bibliographies of books, and abstracts and proceedings of international conferences were reviewed. Randomised controlled trials evaluating change in Hb levels with interventions that included oral or parenteral iron supplementation or iron-fortified formula milk or cereals were analysed. RESULTS: A total of 55 trials (56 cohorts) provided relevant information. Publication bias was evident (P < 0.001). The pooled estimate (random-effects model) for change in Hb with iron supplementation (weighted mean difference) was 0.74 g/dL (95% CI, 0.61-0.87; P < 0.001; P < 0.001 for heterogeneity). Lower baseline Hb level, oral medicinal iron supplementation, and malarial nonhyperendemic region were significant predictors of greater Hb response and heterogeneity. Projections suggested that, on average, between 37.9% and 62.3% of baseline anaemia (Hb <11 g/dL) was responsive to iron supplementation among children under 6 years of age; the corresponding range for malarial hyperendemic regions was 5.8% to 31.8%. CONCLUSIONS: This systematic review indicates that iron supplementation increases Hb levels in children significantly but modestly. The increase is greater in subjects who are anaemic at the start of the trial and lower in malarial hyperendemic areas and in those consuming iron-fortified food. The projected reductions in prevalence of anaemia with iron supplementation alone highlight the need for additional area-specific interventions, particularly in malaria-prone regions. PMID- 17414147 TI - A prospective assessment of food and nutrient intake in a population of Malawian children at risk for kwashiorkor. AB - OBJECTIVE: To determine what foods, nutrients, and dietary patterns are associated with development of kwashiorkor in populations of vulnerable 1- to 3 year-old Malawian children. PATIENTS AND METHODS: This was a prospective observational study conducted in 8 rural villages. Upon enrollment, demographic, anthropometric, and dietary intake data were collected. Children were studied every 2 weeks for 10 weeks to determine whether they developed kwashiorkor. Dietary intake was assessed on enrollment using a food frequency questionnaire, which included all possible foods in the child's diet. Food frequency data were used to estimate energy, protein, vitamins C and A, niacin, thiamin, zinc, and iron intake using food composition and serving size data. Dietary diversity was assessed with a 7-point score. Regression modeling was used to determine whether the consumption of any food or nutrient was associated with the development of kwashiorkor. RESULTS: A total of 43 (2.6%) of the 1651 healthy children ages 1 to 3 years enrolled developed kwashiorkor. Children who developed kwashiorkor were younger and had more nutritional wasting than those who did not. Thirty children (70%) who developed kwashiorkor were breast-fed. In the combined regression model no foods or nutrients were found to be associated with the development of kwashiorkor. There were no differences in the dietary diversity between children who developed kwashiorkor and those who did not. CONCLUSIONS: No association between the development of kwashiorkor and the consumption of any food or nutrient was found. PMID- 17414148 TI - Recurrent intussusception and gastrointestinal bleeding secondary to gastric heterotopia of the small intestine. PMID- 17414149 TI - Eosinophilic esophagitis and gastritis in Rubinstein-Taybi syndrome. PMID- 17414150 TI - Neonatal transient eosinophilic colitis causes lower gastrointestinal bleeding in early infancy. AB - BACKGROUND: Lower gastrointestinal bleeding (LGB), particularly in newborns, is of serious concern and requires urgent investigation and hospital care. Whereas allergic proctocolitis caused by food protein is a significant cause of LGB in infants with eosinophilia, there are several cases of diseases with symptoms similar to those of allergic proctocolitis but without an apparent allergic reaction influence. PATIENTS AND METHODS: We examined 2 neonates using rectosigmoidoscopy who showed eosinophilia and experienced fresh LGB soon after birth and before their first feedings. Serum eosinic cationic protein (ECP) and platelet activating factor (PAF) levels were also examined in the second case to confirm the involvement of eosinophils for its pathogenesis. RESULTS: Both patients were in a clinically stable condition, and their abdomens were soft. The results of their blood analyses, abdominal radiographs, and stool cultures were normal, but they had gross eosinophilia: the eosinophil counts were 9014/mm3 (patient 1) and 1955/mm3 (patient 2). Rectosigmoidoscopy with colonic mucosal biopsy revealed nodular lymphoid hyperplasia with a pale mucosal surface and massive oozing with diffuse eosinophilic infiltration in the lamina propria. In patient 2 the serum ECP and PAF levels were elevated to 123 microg/L (normal, <14.7) and 13.1 micromol/L/min (normal, <6). A few days after intravenous hydration therapy, LGB was no longer detected, and the serum ECP and PAF levels returned to normal. CONCLUSIONS: Inasmuch as these infants had LGB similar to allergic proctocolitis without any allergic reactions, we suggest that infiltrated eosinophils in the colonic mucosa could be involved in the pathogenesis of LGB in early infancy. PMID- 17414151 TI - Gastrointestinal bleeding in children following ingestion of low-dose ibuprofen. PMID- 17414152 TI - Re: Clinical efficacy of probiotics: review of the evidence with focus on children. PMID- 17414154 TI - Re: Acquisition of competence in paediatric ileocolonoscopy with virtual endoscopy training. PMID- 17414155 TI - Measurement of serum acetaminophen-protein adducts in patients with acute liver failure. PMID- 17414156 TI - Wireless capsule endoscopy in the pediatric age group: experience and complications. AB - BACKGROUND: The development of wireless capsule endoscopy (CE) provides a unique opportunity to visualize the entire small bowel in a minimally invasive manner. Studies in adult patients have demonstrated that the disposable capsule is well tolerated and highly effective, but few studies have been done in children. The aims of our study were to compare the diagnostic yield of CE and small bowel series in children being evaluated for possible small intestine disease and to determine the risk of developing an adverse event following capsule endoscopy. PATIENTS AND METHODS: We retrospectively reviewed the records of all children who underwent CE at 1 institution between August 2002 and July 2005. Results of CE were compared with those of small bowel radiographic studies when available. RESULTS: There were 46 CE studies from 45 patients, 28 male and 17 female, with a mean age of 14.9 +/- 3.6 years and mean weight of 49.7 +/- 17.5 kg. The indications for CE included unresponsive Crohn disease (n = 16), possible intestinal polyps (n = 11), unexplained iron deficiency anemia (n = 7), growth failure (n = 5), unresponsive ulcerative colitis (n = 3), persistent abdominal pain (n = 1), protein-losing enteropathy (n = 1), and allergic enteropathy with occult gastrointestinal bleeding (n = 1). Of the 46 CE studies, 41 were completed and 5 were incomplete studies. Based on the CE, 9 patients were newly diagnosed with Crohn disease, 9 patients with Crohn disease were newly diagnosed with small bowel involvement, 8 patients had upper intestinal polyps, 1 patient had findings consistent with Menetrier disease, and 1 had a duodenal ulcer. Thirty-three patients had small bowel series before CE: 24 studies were normal, 6 had abnormal thickening of the small bowel, 2 had polyps, and 1 patient had antral narrowing. All 9 patients with abnormal small bowel series had abnormal CE studies. Of the 24 patients with normal small bowel series, 20 had completed CE studies, and in 10 children, the study was abnormal. Nine of the 45 subjects had adverse events. Five patients had delayed passage from the stomach, with 2 needing endoscopic retrieval of the CE, and 4 had delayed passage from the small intestine (>5 days), with 2 requiring surgical removal, 1 responding to steroids, and the final patient requiring an ileocolic resection 2 months after the CE for an undiagnosed ileal stricture. The only significant association noted was that older patients were more likely to have intestinal retention. CONCLUSIONS: CE provides a valuable tool in the evaluation of pediatric patients for possible small bowel disease. However, the risk of developing complications appears to be greater in the pediatric population, with 20% of our patients having an adverse event. PMID- 17414157 TI - Removal of impacted food in children with eosinophilic esophagitis using Saeed banding device. AB - Food impaction in patients with eosinophilic esophagitis is being recognized increasingly. Retrieval of impacted food may be necessary if it does not resolve spontaneously. The present report describes a new technique for removal of friable impacted food from the esophagus using a modified version of the Saeed banding device. With this technique, impacted food was safely removed in a single attempt in each of 3 patients. PMID- 17414158 TI - Community-based case-control study of childhood chronic abdominal pain: role of selected laboratory investigations. AB - OBJECTIVE: To compare laboratory investigation results in children with chronic abdominal pain and in healthy control children. Our hypothesis was that parasitic infection was not a causal factor for chronic abdominal pain and that there would be no difference in leukocyte count, hemoglobin level, or erythrocyte sedimentation rate (ESR) between the 2 groups. PATIENTS AND METHODS: Children with chronic abdominal pain and healthy control children (5-15 years) were recruited from the practices of 6 primary care pediatricians in Toronto, Canada. Stool samples were analyzed for ova and parasites, and serum samples were used to estimate leukocyte count, hemoglobin, and ESR. A standardized questionnaire was used to gather social, demographic, and clinical information. RESULTS: A total of 157/200 children (79%) provided samples. Children with chronic abdominal pain were more likely to be female than were control children. Stool samples were positive for parasitic infection in 15 children, with no difference in prevalence between children with chronic abdominal pain (6/87; 7%) and healthy control children (9/70; 13%); P = 0.28). The mean (standard deviation) leukocyte count in children with chronic abdominal pain was 7.4 x 10(9)/L (2.03), compared with 8.3 x 10(9)/L (1.82) in healthy control children. No child had a leukocyte count above 20 x 10(9)/L. The mean (SD) hemoglobin in children with chronic abdominal pain was 131 g/L (8.4), compared with 130 g/L (9.2) in healthy control children. Last, the median ESR in children with chronic abdominal pain was 5 mm/hour, compared with 3 mm/hour in control children. CONCLUSIONS: The study findings suggest that in the absence of alarming symptoms and signs, parasitic infection is not a causal factor for chronic abdominal pain and that routine screening tests (leukocyte count, hemoglobin, ESR) are not useful. PMID- 17414160 TI - Detection of a transcript abnormality in mRNA of the SLC12A3 gene extracted from urinary sediment cells of a patient with Gitelman's syndrome. AB - To date, many mutations, including intronic nucleotide changes, in the SLC12A3 gene encoding the thiazide-sensitive sodium-chloride cotransporter (NCCT) have been reported in Gitelman's syndrome (GS) patients. However, it has not been clarified whether intronic nucleotide changes affect mRNA content. Since mRNA analysis is possible only after obtaining renal biopsy specimens, no studies have been conducted to identify transcript abnormalities in GS. In the study reported here, we investigated such transcript abnormalities for the first time by using mRNA expressed in a patient's urinary sediment cells. Direct sequencing analysis of leukocyte DNA disclosed one known missense mutation (R399C) and one known nucleotide change of the splicing acceptor site of intron 13 (1670-1 g > t). mRNA extracted from the urinary sediment cells was analyzed by RT-PCR to determine the pathogenic role of the intron mutation. A fragment encompassing exon 13 to 15 was amplified as two products, one consisting of all three exons and the other lacking only exon 14 in its entirety. Our investigation was the first to demonstrate exon 14 skipping in an NCCT transcript in renal cells. This methodology thus constitutes a potential noninvasive analytical tool for every inherited kidney disease. PMID- 17414161 TI - Reconstruction of bone defects using rhBMP-2-coated devitalized bone. AB - Massive bone defects often are caused by radical resection of bone tumors. Reconstruction of the defect by reimplantation of the resected bone segment after it has been devitalized is advantageous because of its ability to match the size of the defect. In addition, this technique carries a low risk for local recurrence of the tumor, avoids immunologic reaction, and is low in cost. However, limited osteogenic potential of the devitalized bone often leads to delayed union, gradual resorption, and mechanical weakness of the reimplanted segment. We applied rhBMP-2 in a biodegradable polymer delivery system to the devitalized bone. Middiaphyseal bone defects were created by resection in rat femurs. The resected segments were autoclaved at 135 degrees C for 15 minutes, coated with a rhBMP-2-retaining paste on the outer surface, and then reimplanted into the defects. In a brief time, newly formed bone was seen on the surface of the devitalized bone. After 12 weeks, a solid bone mass encasing the dead bone segments was consistently formed and abundant new bone formation was visible in the segments as they were remodeled. The amount of new bone formed could be regulated by the amount of the rhBMP-2-retaining paste applied to the bone segments. This method presents a new approach for the reconstruction of bone defects. PMID- 17414162 TI - A new technique for physiodesis using photodynamic therapy. AB - Vascularization of developing growth plates is integral to the process of endochondral ossification. We hypothesized photodynamic therapy could be used to initiate premature vascularization and calcification of growth plates in mice on the induction of vascular endothelial growth factor. Three-week-old transgenic mice that emit bioluminescence on activation of the vascular endothelial growth factor gene were treated with different regimens of benzoporphyrin derivative mono-acid-mediated photodynamic therapy in the right, proximal tibial growth plate. We quantified changes in vascular endothelial growth factor-related bioluminescence at times after photodynamic therapy in vivo. The expression of vascular endothelial growth factor protein and CD31-labeled vasculature in growth plates also were examined with growth plate histology. Measurements of limb length were assessed in vivo using conventional radiography and confirmed on harvesting. Mice exposed to repeat treatments of 10 J x2 displayed enhanced bioluminescence 2 weeks after photodynamic therapy. Histology confirmed increased vasculature immediately adjacent to the growth plates with evidence of physeal closure. At 4 weeks posttreatment, limbs were shortened by an average of 9.5% +/- 4.4% without complication, confirming the potential application of photodynamic therapy for physiodesis. PMID- 17414163 TI - Femoral neck fractures: can physiologic status determine treatment choice? AB - In patients with displaced femoral neck fractures, meta-analysis data show revision rates of 35% after internal fixation and 16% after hemiarthroplasty. A published physiologic status score management protocol, which selects for either treatment, suggests lower revision rates can be achieved but it has not been confirmed. The physiologic status score included subscores for mobility, accommodation, bone density, cognition, and American Society of Anesthesiologists class. We asked whether treatment selection with the physiologic status score could indeed reduce revision rates compared with meta-analysis data and whether surgical technique influenced results. In a prospective multicenter 2-year followup trial we enrolled 115 patients selected for internal fixation and 109 for hemiarthroplasty, aged 60 to 90 years. Healthier mobile patients underwent internal fixation and patients with a lower physiologic status score had hemiarthroplasty. The 2-year revision rate was 40% after internal fixation (above meta-analysis rate) and 3% after hemiarthroplasty (below meta-analysis rate). After a panel identified and excluded 15 technical failure cases, patients younger than 80 years with a high physiologic status score had a one in four revision rate of internal fixation, whereas older patients had a one in two revision rate. Although the revision rate was indeed low after hemiarthroplasty, treatment choice based on physiologic status does not substantially improve clinical decision making. PMID- 17414164 TI - Relationship between femoral head size and distance to lesser trochanter. AB - Hemiarthroplasty of the hip is a common treatment for displaced fractures of the femoral neck. However, accurate restoration of leg length may be difficult to achieve because intraoperative measurements of leg length and preoperative templating are limited by the fracture. Our primary question was whether femoral head diameter can be used to predict femoral head height. We analyzed 34 cadaveric femora to determine diameter of the femoral head, distance from the lesser trochanteric sulcus to the external center of the femoral head (head height), lateral distance from the proximal tip of the greater trochanter to the center of the femoral head (offset), and the femoral neck/shaft angle. The femoral head diameter had a linear correlation with head height expressed by the following equation: head height = 1.035 x femoral head diameter. Our results indicate the diameter of the femoral head is similar in size to the distance from the top of the lesser trochanter to the center of the femoral head in an average population of cadavers. This may be a helpful clinical measurement to assess leg length during surgical treatment of hip fractures. PMID- 17414165 TI - Treatment of tuberculosis of the cervical spine: operative versus nonoperative. AB - We retrospectively reviewed 15 children (four with paralysis) and 39 adults (10 with paralysis) with tuberculosis of the cervical spine to assess the drug responses, disease arrest, and healing times. Ten children and 13 adults were treated nonoperatively, while anterior debridement was performed in five children (two with paralysis) and anterior radical surgery in 26 adults (10 with paralysis). Triple chemotherapy (isoniazid, rifampin, ethambutol [or pyrazinamide for children]) was given to all patients for 12 months. The tuberculosis began to arrest after 3 months of chemotherapy and healed within 12 months. Spontaneous fusion occurred in all adults but only two of the 10 children. Surgical fusion was achieved within 12 to 16 weeks in adults. In nonoperated patients, an initial kyphosis of 12 degrees progressed to 17 degrees at final followup in the children and an initial kyphosis of 9 degrees progressed to 13 degrees in 13 adults. In operated patients, the initial kyphosis of 13 degrees in adults became 2 degrees at the time of the fusion, while the initial kyphosis of 14 degrees in the five children progressed to 18 degrees. Patients with paraplegia recovered completely within 14 days on average (range, 1-42 days) after treatment. Recovery was gradual in the nonoperative group, while it occurred within 3 days in the operative group. PMID- 17414166 TI - Osteosarcoma, chondrosarcoma, and Ewing's sarcoma: National Cancer Data Base Report. AB - We summarize descriptive epidemiologic and survival data from the National Cancer Data Base of the American College of Surgeons for 26,437 cases of osteosarcoma (n = 11,961), chondrosarcoma (n = 9606), and Ewing's sarcoma (n = 4870) from 1985 to 2003. Survival data are reported on cases with a minimum 5-year followup from 1985 to 1998 (8,104 osteosarcomas, 6,476 chondrosarcomas, and 3,225 Ewing's sarcomas). The relative 5-year survival rate was 53.9% for osteosarcoma, 75.2% for chondrosarcoma, and 50.6% for Ewing's sarcoma. Survival rates did not change notably over the collection period. Within osteosarcomas, the relative 5-year survival rates were 52.6% for high grade, 85.9% for parosteal, and 17.8% for Paget's subtypes. For osteosarcoma patients, the relative 5-year survival rate was 60% for those younger than 30 years, 50% for those aged 30 to 49 years, and 30% for those aged 50 years or older. Within chondrosarcomas, the relative 5-year survival rate was 76% for conventional, 71% for myxoid, 87% for juxtacortical, and 52% for mesenchymal. While the National Cancer Data Base has limitations, the survival data and demographics for bone sarcomas are unprecedented in numbers and duration. Our report supports continued efforts to refine data collection and stimulate further data analysis. PMID- 17414167 TI - Oncogene coexpression in mesenchymal neoplasia correlates with EGF transcription. AB - Epidermal growth factor (EGF) is a potent mitogenic factor for cells of mesodermal and ectodermal origin, and its over-expression is associated with a variety of cancers. We asked whether oncogene coexpression occurs in mesenchymal neoplasms, if coexpression correlates with EGF transcription, and whether coexpression can be attributed to the EGF-induced overexpression of oncogenes. We quantified the mRNA concentrations of EGF and 14 oncogenes in 42 primary sarcomas, 31 benign tumors, and 10 skeletal muscle controls, and compared mRNA concentrations and gene pair correlations in EGF positive (EGF+) tumors to transcript concentrations and correlations in EGF negative (EGF-) tumors. Transcripts were detected by real time reverse transcription-polymerase chain reaction. Pearson's correlation coefficients identified gene associations, and gene synchrony associated with EGF expression was evaluated using chi square. Transcript concentrations in tumors were compared graphically and with t tests. Gene correlations predominated in EGF+ benign tumors and in EGF- primary sarcomas. The dichotomy in oncogene coexpression evident in benign and malignant tumors could not be attributed to statistical differences in mRNA content between EGF+ and EGF- tumors. EGF may enhance, or may indicate the presence of, oncogene coexpression in benign mesenchymal lesions, but counters gene synchronization in sarcomas. PMID- 17414168 TI - Rotationplasty for bone tumors: is there still a role? AB - We evaluated the disease status and functional results in 30 patients (range, 6 25 years) who underwent rotationplasty for bone sarcomas from January 2000 to February 2004. Plating was used for fixation in all 27 distal femur resections. In the proximal femur tumor, the distal femur was contoured and fixed to the ilium with cancellous screws. In the two cases involving the entire femur, the upper end of the tibia was articulated with the acetabulum in one case and an Austin Moore prosthesis was inserted in the upper end of the tibia in the other. Two patients underwent an amputation after postoperative vascular compromise. Other complications included venous congestion in two patients, who recovered after exploration, partially recovered nerve palsy in one, wound infection in two, local recurrence in one, and nonunion with subsequent bone grafting in one. Functional evaluation was documented in 26 patients with a followup ranging from 24 to 60 months. Using the Musculoskeletal Tumor Society scoring system, the score was 25 or greater in 20 of 26 patients. In appropriately indicated cases, rotationplasty provides good local disease control and good function. PMID- 17414169 TI - Late radial head dislocation with radial head fracture and ulnar plastic deformation. AB - Type II Monteggia lesion equivalents produced by plastic deformation of the ulna are rare. Radial head fractures in skeletally immature patients are also uncommon. We report a late presentation of a Type II Monteggia equivalent injury with a fracture of the radial head and neck and plastic deformation of the ulna in an 11-year-old boy. The radial head was located on the initial injury radiographs and subsequently dislocated in a posterior direction. The radial head fracture was misdiagnosed as a coronoid fracture at presentation. The plastic deformation of the ulna was diagnosed several weeks after the injury when the dislocation was first noted. Magnetic resonance imaging was used to establish the diagnosis of a radial head fracture at the same time. The patient was treated successfully with an open reduction and internal fixation of the radial head combined with a dorsal closing-wedge ulnar osteotomy. PMID- 17414170 TI - Range of motion correlates with patient perception of TKA outcome. AB - Despite the clinical success of total knee arthroplasty, little information can be found in the literature about the relationship between certain postoperative physical findings and the outcome. Specifically, is the range of motion related to patient perception of outcome? We performed a cohort prospective study on 48 patients assessed by patient-oriented evaluations (Short Form 36 Health Survey and Oxford Knee Questionnaire) and objective evaluations after total knee arthroplasty. Thirty-four patients were women and 14 were men. The mean age at followup was 71 years (range, 64-80 years) and the minimum followup was 20 months (mean, 28.5 months; range, 20-30 months). We found a positive correlation between range of motion and patient-oriented evaluations in some domains of the Short Form 36 and in the Oxford knee score. PMID- 17414171 TI - Mammographic density in a multiethnic cohort. AB - OBJECTIVES: To compare mammographic density among premenopausal and early perimenopausal women from four racial/ethnic groups and to examine density and acculturation among Japanese and Chinese women. DESIGN: The study included 391 white, 60 African American, 171 Japanese, and 179 Chinese participants in the Study of Women's Health Across the Nation, a multisite study of US women transitioning through menopause. Mammograms done when women were premenopausal or early perimenopausal were assessed for area of dense breast tissue and the percent of the breast occupied by dense tissue (percent density). Information on race/ethnicity, acculturation, and other factors was obtained from standardized instruments. Multiple linear regression modeling was used to examine the association between race/ethnicity or acculturation and density measures. RESULTS: Age-adjusted mean percent density was highest for Chinese (52%) and lowest for African American (34%) women. After additional adjustment for body mass index, menopause status, age at first birth, breast-feeding duration, waist circumference, and smoking, African Americans had the highest mean percent density (51%) and Japanese women had the lowest (39%). In contrast, the area of dense tissue was highest for African Americans and similar for white, Japanese, and Chinese women. Less acculturated Chinese and Japanese women tended to have a larger area of density and a higher percent density. CONCLUSIONS: Neither the age adjusted nor fully adjusted results for percent density or area of dense tissue reflected current differences in breast cancer incidence rates among similarly aged African American, Japanese, Chinese, and white women. In addition, mammographic density was higher in less acculturated Asian women. PMID- 17414174 TI - Hydroxyapatite cement cranioplasty in translabyrinthine acoustic neuroma surgery update. AB - OBJECTIVE: To update the outcome of hydroxyapatite cement cranioplasty in translabyrinthine acoustic neuroma (TLAN) surgery. PATIENTS: One hundred eight previously reported patients undergoing abdominal fat graft reconstruction versus hydroxyapatite cement cranioplasty with additional 4-year follow up. Ninety additional patients undergoing a uniform technique of hydroxyapatite cement cranioplasty after TLAN. INTERVENTION(S): After TLAN, strips of abdominal fat are placed through the dural opening and medial to the level of the mastoid antrum, filling the lateral mastoid cavity. MAIN OUTCOME MEASURE(S): Cerebrospinal fluid (CSF) leaks and wound complications. RESULTS: No additional CSF leaks or wound complications were identified in the patients included in the previous report. In the new series of 90 consecutive patients, there was one CSF leak. CONCLUSION: Hydroxyapatite cranioplasty is a reliable method to avoid CSF leaks after TLAN surgery. PMID- 17414177 TI - CPA melanoma: diagnosis and management. AB - OBJECTIVE: Melanoma rarely invades the cerebellopontine angle (CPA) and can evade accurate diagnosis, which may alter management decisions. Diagnosis may be facilitated via careful history, magnetic resonance imaging (MRI) findings, and cerebrospinal fluid (CSF) analysis. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: Thirteen internal auditory canal/CPA lesions in eight patients who presented with CPA syndrome and who had a pathological diagnosis consistent with malignant melanoma. There were four bilateral and four unilateral lesions. Six of eight patients had a history of melanoma. One was apparently primary CPA lesion, whereas all others were metastatic. INTERVENTION(S): T1- and T2-weighted precontrast and postcontrast gadolinium-enhanced MRI were obtained, including fat suppression and fluid attenuated inversion recovery sequence images in two patients; lumbar puncture with CSF centrifugation and cytological analysis confirmed the diagnosis in two patients. Translabyrinthine craniotomy was performed for tumor extirpation in five patients. MAIN OUTCOME MEASURE(S): Symptoms at presentation, MRI findings, presence of malignant cells in CSF, tumor progression, intraoperative findings, response to treatment, time interval from initial diagnosis of melanoma elsewhere, and survival. RESULTS: Seven of eight patients had history and/or MRI findings suggestive of malignancy in the internal auditory canal and/or CPA, and diagnosis was confirmed via CSF analysis in two patients. In one patient, diagnosis was made at surgery. CONCLUSION: Internal auditory canal melanoma portends a grim prognosis, can occur up to 17 years after initial melanoma diagnosis/treatment, and can be detected with appropriate MRI sequences, especially enhanced fluid-attenuated inversion recovery images. In disseminated cases, diagnosis can be confirmed with lumbar puncture demonstrating malignant cells. Management includes tumor resection when melanoma seems to be solitary and malignant cells are not present in CSF. Intrathecal chemotherapy and radiation are recommended for dissemination, although the survival rate is still poor. PMID- 17414178 TI - Restoration of brain stem auditory-evoked potential in maple syrup urine disease. AB - OBJECTIVE: The purpose of this study was to report a unique case of reversible brain stem auditory-evoked potential (BAEP) involving wave I in maple syrup urine disease (MSUD). The possible mechanism of the hearing loss and reversibility of wave I is proposed. PATIENTS: Single case report. INTERVENTION: Treatment of MSUD with dialysis and diet devoid in branched-chain amino acids. MAIN OUTCOME MEASURE: BAEPs. RESULTS: Initial presentation of MSUD in a 14-week-old child included failure to thrive, present otoacoustic emissions, and absent BAEPs. After treatment with dialysis and an MSUD-appropriate diet, the BAEPs gradually recovered. CONCLUSION: Appropriate and timely treatment of MSUD can reverse the central neuropathy leading to hearing loss. Because patients with MSUD have elevated levels of glutamate, we propose that the auditory dysfunction in these patients may be related to glutamate excitotoxicity, and with appropriate treatment, these effects may be reversible. PMID- 17414179 TI - Atypical clinical presentations of vestibular schwannomas. AB - A significant number of patients with vestibular schwannomas present atypically, with none of the classical symptoms of unilateral sensorineural hearing loss, tinnitus, and/or dysequilibrium. The aim of this study is to highlight those patients with unusual clinical symptoms. STUDY DESIGN: The clinical data of all patients who presented to the vestibular schwannoma clinic at Beaumont Hospital over the past 12 years was prospectively recorded in a computerized database. This paper reviews the atypical presenting symptoms. RESULTS: Three hundred ninety-eight patients were included in this study. A total of 3.7% of patients presented with atypical symptoms only. CONCLUSION: A significant subgroup, 3.7% in our study, did not present with the audiovestibular symptoms classically associated with vestibular schwannoma. Clinician awareness of the atypical clinical symptoms may lead to earlier detection of these lesions. PMID- 17414180 TI - Comparison of hearing results of nitinol SMART stapes piston prosthesis with conventional piston prostheses: postoperative results of nitinol stapes prosthesis. AB - OBJECTIVE: Recently, a new stapedotomy piston prosthesis, which is a composite metal alloy of nickel and titanium known as nitinol, has been introduced into medical use. This biocompatible alloy has the unique property of shape-memory, which permits tight self-crimping when heat is applied to the wire. To substantiate the favorable initial observations with the SMART piston, this study was undertaken to compare these results (n=26) with those obtained using conventional stainless steel or platinum ribbon prostheses (n=28). STUDY DESIGN: Prospective consecutive case review: consecutive cases performed by the same surgeon were analyzed. SETTING: Tertiary referral center. PATIENTS: Fifty-four healthy patients with otosclerosis. INTERVENTION: Stapedotomy using either SMART prosthesis or conventional prosthesis. MAIN OUTCOME MEASURES: Hearing outcomes by audiological assessment. RESULTS: The postoperative hearing mean pure-tone average was 24.81+/-16.20 dBHL for Group 1 (SMART prosthesis) and 27.46+/-15.57 dBHL for Group 2 (conventional prosthesis). Postoperative mean air-bone gap was 7.07+/-8.14 dBHL for Group 1 and 6.38+/-7.54 dBHL for Group 2 using 0.5-, 1-, 2-, and 4-kHz frequencies. When analyzed according to the American Academy of Otolaryngology-Head & Neck Surgery reporting criteria using an estimate of 3 kHz as a mean of the 2-and 4-kHz values, the postoperative mean air-bone gap was 5.42+/-5.4 dBHL for Group 1 SMART and 5.98+/-5.47 dBHL for Group 2 conventional prostheses. Postoperative speech discrimination scores were 96%+/-8.64% and 97%+/ 5.9%, respectively. These differences were not shown to be statistically different. CONCLUSION: Results demonstrate that experienced surgeons may achieve comparable results with both prostheses; however, the ease of self-crimping and the tightness of the crimp may provide advantages that may have long-term benefits. The potential issue of nickel allergy is important when considering patients for this prosthesis. PMID- 17414181 TI - Loss of stapes prosthesis into vestibule during stapedectomy: a case report. AB - We present a unique case of intraoperative loss of a piston prosthesis into the right vestibule of a 47-year-old man during stapedectomy. Despite the fact that the prosthesis remains unfixed in the scala vestibuli at the basal turn of the cochlea, the patient has remained nearly asymptomatic in his right ear for approximately 2 years. We discuss the patient's presenting condition, operative course, and our diagnostic and management strategy in this case. PMID- 17414182 TI - Important factors for the hearing loss caused by the triple semicircular canal occlusion in guinea pigs. AB - HYPOTHESIS: Dynamic recording of the auditory brainstem response is helpful in verifying harmful procedure(s) to hearing during triple semicircular canal occlusion (TSCO) surgery. The damage to the membranous semicircular labyrinth is the single major contributor to hearing loss caused by TSCO. BACKGROUND: Posterior semicircular canal occlusion has been recognized as an efficient method of eliminating vertigo without causing a significant hearing impairment. Recently, TSCO has also been explored for its potential to treat vertigo of various causes. In limited animal studies, varied hearing impairments have been documented after TSCO. However, the major factor(s) causing hearing loss in TSCO is/are unclear. METHODS: Triple semicircular canal occlusion was performed on 36 guinea pigs in total. The cochlear function of the guinea pigs was monitored by observing the auditory brainstem response. The impact of membranous labyrinth damage on hearing was verified by a between-group comparison. RESULTS: Hearing loss during TSCO was accumulated in every step of semicircular canal manipulation. Generally, perilymph leak was found to cause a slight hearing loss that was predominately recovered during surgery. However, transaction of the membranous labyrinth usually caused a more significant hearing loss that was not recovered during the surgery. In addition, the magnitude of hearing loss seemed to be increased with the elongation of the surgery. However, the hearing can be largely recovered after the surgery even in animals with transaction of the membranous labyrinth. CONCLUSION: Hearing loss caused by TSCO can be greatly reduced by avoiding damage to the membranous labyrinth and by shortening the operation time. PMID- 17414184 TI - Values and ethics: a collection of curricular reforms for a new generation of physicians. AB - In recent years, medical educators have expressed concern that the reductionist positivist mode of medical education fails to equip physicians with the skills and attitudes to meet the full range of patients' physical and emotional needs. Indeed, the authors suggest that neither patients nor physicians are satisfied. Among the factors responsible are a pervasive industrialization of clinical practice, a progressive segmentation of patient care, and a deepening shortage of both primary care and specialty physicians. But underlying these system issues is a lack of adequate schooling in the values, ethics, and culture of caring. Today's physicians must simultaneously be analytical, perceptive, and self reflective. They must have the capacity to see their patients as individuals with differing psychological, social, and historical natures. And they must have insight into their own values and behaviors. All of this contributes to making a competent and humane physician. To aid medical students in achieving these characteristics, the authors contend that medical education must be radically restructured so that knowledge and skills are taught within the context of values and ethics. This commentary explores such reform through the lens of three articles published in the current issue of Academic Medicine, by Litzelman and Cottingham, Kanter and colleagues, and Dobie. These articles are the product of a national call that resulted in more than thirty abstracts, testimony to the fertile thinking already being applied to this problem. It is the authors' hope that this series of papers will stimulate still more thinking and lead to the curricular reform that future generations of physicians deserve. PMID- 17414186 TI - Leadership lessons from curricular change at the University of California, San Francisco, School of Medicine. AB - After successive Liaison Committee on Medical Education accreditation reports that criticized the University of California, San Francisco, School of Medicine for lack of instructional innovation and curriculum oversight, the dean issued a mandate for curriculum reform in 1997. Could a medical school that prided itself on innovation in research and health care do the same in education? The authors describe their five-phase curriculum change process and correlate this to an eight-step leadership model. The first phase of curricular change is to establish a compelling need for change; it requires leaders to create a sense of urgency and build a guiding coalition to achieve action. The second phase of curriculum reform is to envision a bold new curriculum; leaders must develop such a vision and communicate it broadly. The third phase is to design curriculum and obtain the necessary approvals; this requires leaders to empower broad-based action and generate short-term wins. In the fourth phase, specific courses are developed for the new curriculum, and leaders continue to empower broad-based action, generate short-term wins, consolidate gains, and produce more change. During the fifth phase of implementation and evaluation, leaders need to further consolidate gains, produce more change, and anchor new approaches in the institution. Arising from this experience and the correlation of curricular change phases with leadership steps, the authors identify 27 specific leadership strategies they employed in their curricular reform process. PMID- 17414187 TI - Lessons from industry: one school's transformation toward "lean" curricular governance. AB - As medical education grapples with organizational calls for centralized curricular oversight, programs may be compelled to respond by establishing highly vertical, stacked governance structures. Although these models offer discrete advantages over the horizontal, compartmentalized structures they are designed to replace, they pose new challenges to ensuring curricular quality and the educational innovations that drive the curricula. The authors describe a hybrid quality-assurance (QA) governance structure introduced in 2003 at the University of Kentucky College of Medicine (UKCOM) that ensures centralized curricular oversight of the educational product while allowing individualized creative control over the educational process. Based on a Lean production model, this approach draws on industry experiences that strategically separate institutional accountability (management) for a quality curriculum from the decision-making processes required to ensure it (production). In so doing, the authors acknowledge general similarities and key differences between overseeing the manufacture of a complex product versus the education of a physician-emphasizing the structured, sequential, and measurable nature of each process. Further, the authors briefly trace the emergence of quality approaches in manufacturing and discuss the philosophical changes that accompany transition to an institutional governance system that relies on vigorous, robust performance measures to offer continuous feedback on curricular quality. PMID- 17414188 TI - Creating sustainable curricular change: lessons learned from an alternative therapies educational initiative. AB - The authors describe the process by which a curriculum was developed to introduce complementary and alternative medicine topics at multiple levels from health professional students to faculty, as part of a five-year project, funded by a grant from the National Institutes of Health, at the University of Texas Medical Branch in Galveston, Texas, from 2001 to 2005. The curriculum was based on four educational goals that embrace effective communication with patients, application of sound evidence, creation of patient-centered therapeutic relationships, and development of positive perspectives on wellness. The authors analyze the complex and challenging process of gaining acceptance for the curriculum and implementing it in the context of existing courses and programs. The developmental background and context of this curricular innovation at this institution is described, with reference to parallel activities at other academic health centers participating in the Consortium of Academic Health Centers for Integrative Medicine. The authors hold that successful curricular change in medical schools must follow sound educational development principles. A well-planned process of integration is particularly important when introducing a pioneering curriculum into an academic health center. The process at this institution followed six key principles for successful accomplishment of curriculum change: leadership, cooperative climate, participation by organization members, politics, human resource development, and evaluation. The authors provide details about six analogous elements used to design and sustain the curriculum: collaboration, communication, demonstration, evaluation, evolution, and dissemination. PMID- 17414190 TI - Development of eMed: a comprehensive, modular curriculum-management system. AB - In 2001 the University of New South Wales Faculty of Medicine embarked on designing a curriculum-management system to support the development and delivery of its new, fully integrated, outcome-based, six-year undergraduate medicine program. The Web-enabled curriculum-management system it developed is known as eMed, and it comprises a suite of integrated tools used for managing graduate outcomes, content, activities, and assessment in the new program. The six main tools are a curriculum map, timetable, student portfolio, peer feedback tool, assessment tracking, and results tools. The eMed functions were determined by organizational and curricular needs, and a business management perspective guided its development. The eMed project was developed by a multidisciplinary team, and its successful development was achieved mostly by methodically identifying the scope of each tool and the business processes it supports. Evaluation results indicated a high level of user acceptance and approval. The eMed system is a simple yet effective educational technology system that allows users to evaluate and improve the curriculum in real time. As a second-generation curriculum management system, eMed is much more than an educational administration system; it is a knowledge network system used by staff and students to transform data and information into knowledge and action. The integration of learning and assessment activities data in the one system gives a depth of curriculum information that is unusual and that allows for data-based decision making. Technologically, eMed helps to keep the medicine program up to date. Organizationally, it strengthens the school's data-driven decision-making process and knowledge network culture. PMID- 17414192 TI - The University of Missouri-Kansas City School of Medicine: thirty-five years of experience with a nontraditional approach to medical education. AB - The University of Missouri-Kansas City (UMKC) School of Medicine is a public medical school that opened in 1971 in response to a need to train more physicians in Missouri. As a six-year, integrated, combined-degree program leading to the baccalaureate and medical degrees, the school offers an innovative, nontraditional approach to medical education. In the past 35 years, UMKC has graduated over 2,400 physicians who are successful according to outcomes measures used at other medical schools. With recent interest in reforming medical education to prepare physicians for a changing world, a review of alternative models may be especially instructive.UMKC's academic plan offers a blueprint for the curriculum plan and governance of the school. The plan is built on four hallmarks: (1) a combined baccalaureate/MD program, (2) early exposure to clinical medicine, (3) small-group learning through the docent system, and (4) a continuing ambulatory care clinic experience for four years. This article catalogs the results of this plan including student, faculty, and graduates' perceptions of and satisfaction with the school's educational approach, students' achievement on licensing examinations and in the residency match, graduates' performance in residency programs, and their subsequent career patterns. The authors also discuss lessons learned and adjustments made in response to local needs in the context of a changing environment in education, health care, and health care delivery while continually improving the school's nontraditional approach to medical education. These include changes in basic and clinical science instruction, student assessment, faculty development, and funding and governance. PMID- 17414193 TI - PBL in the undergraduate MD program at McMaster University: three iterations in three decades. AB - When the undergraduate MD program of McMaster University admitted its first cohort of 20 students in 1969, it heralded a major change in medical school pedagogy that has influenced the education of medical students around the world. The three-year PBL curriculum, which emphasized small-group tutorials, self directed learning, a minimal number of didactic presentations, and student evaluation that was based almost entirely on performance in the tutorial, represented a radical departure from traditional curricula. Since the inception of the original curriculum in 1969, there have been two major curriculum revisions, the most recent of which was in 2005. The original curriculum attempted to integrate both basic science and clinical science into the biomedical problems. The second iteration of the curriculum focused on priority health problems and centered on a list of common medical problems as the foundation for curriculum organization, on the basis that an understanding of the management of common conditions included areas of knowledge that would be essential for clinical competence. Under the third, current curriculum, the COMPASS (concept-oriented, multidisciplinary, problem-based, practice for transfer, simulations in clerkship, streaming) model was adopted. Under this concept-based system, emphasis is placed on underscoring the underlying concepts in the curriculum with a logical sequencing of both the concepts and the body systems. This article briefly reviews the history of the development of the undergraduate MD program at McMaster and the three curricula that have been developed during the past three decades. PMID- 17414195 TI - A curricular model for the training of physician scientists: the evolution of the Duke University School of Medicine curriculum. AB - Duke University School of Medicine offers an unusual doctor of medicine educational program. The core basic sciences are taught in year one, core clinical clerkships are completed in the second year, the entire third year is devoted to scholarly investigation, and elective rotations are fulfilled in the fourth year. The creation of this unique structure presented many challenges and is the product of a desire of key faculty 40 years ago to change radically the way medical education was taught. Over the years, improvements have been made, but the underlying principles of these visionary leaders have been retained: inquire not just acquire, flexibility of choice, and in-depth exploration. In the spirit of innovation that was established 40 years ago, leaders and faculty at Duke developed a new curricular model in 2004, called Foundation for Excellence, which is anchored in integrated, interdisciplinary innovation. The authors describe the process of curricular reform and provide a detailed overview of this unique approach to medical education. In keeping with Duke's mission to graduate clinician-researchers and clinician-educators, reducing the basic science curriculum to one year created a year saved, which students are now required to devote to scholarly pursuits. The authors argue that adopting a similar one-year basic science curriculum would make instructional time available for other schools to achieve their own institutional goals. PMID- 17414196 TI - Establishing a regional medical campus in southeast Florida: successes and challenges. AB - In August 2007, the first class of University of Miami Miller School of Medicine (UM) medical students will begin the four-year undergraduate medical education program at the regional medical campus at Florida Atlantic University (FAU) The authors describe how UM and FAU were able to make a successful case to state policymakers for a regional medical campus as a cost-effective approach to expanding undergraduate and graduate medical education opportunities in southeast Florida The authors discuss what motivated UM and FAU to partner to create a regional medical campus, and they describe the challenges that have been encountered since 2004, particularly those relating to delivering a comparable two-year program on two campuses using distance-learning technologies. The opportunities that have resulted from expansion of the regional campus from two to four years are also described, including the development of a new and innovative four-year curriculum emphasizing comprehensive chronic disease management and case-based and patient-centered education using collaborative, small-group student learning communities. UM medical students thus have a choice between two educational tracks. The authors conclude that no significant impediments have resulted from the Florida collaboration between a public and a private university and that the regional medical campus model can serve as a viable option for other states and institutions attempting to expand medical school enrollment and meet physician workforce needs in an efficient and cost effective manner. PMID- 17414197 TI - Cleveland Clinic Lerner College of Medicine: an innovative approach to medical education and the training of physician investigators. AB - Cleveland Clinic Lerner College of Medicine (CCLCM) is an innovative, five-year medical education track within Case Western Reserve University School of Medicine (Case) with a focused mission to attract and educate a limited number of highly qualified persons who seek to become physician investigators. CCLCM curriculum governance, faculty appointments and promotions, and admissions committees are integrated with respective Case committees. The CCLCM curriculum is based on faculty-defined professional attributes that graduates are expected to develop. These attributes were used to create curricular and assessment principles that guided the development of an integrated basic science, clinical science, and research curriculum, conducted in an active learning environment. An organ-system approach is used to solidify an understanding of basic science discipline threads in the context of relevant clinical problems presented in PBL and case-based discussion formats. Clinical skills are introduced in the first year as part of the two-year longitudinal experience with a family practice or internal medicine physician. The research program provides all students with opportunities to learn and experience basic and translational research and clinical research before selecting a research topic for their 12- to 15-month master-level thesis project. All Case students participate in required and elective clinical curriculum after the second year, but CCLCM students return to the Cleveland Clinic on selected Friday afternoons for program-specific research and professionalism-learning activities. A unique portfolio-based assessment system is used to assess student achievements in nine competency areas, seven of which reflect the Accreditation Council for Graduate Medical Education competencies. PMID- 17414198 TI - The Harvard Medical School-Cambridge integrated clerkship: an innovative model of clinical education. AB - The Harvard Medical School-Cambridge Integrated Clerkship (HMS-CIC) is a redesign of the principal clinical year to foster students' learning from close and continuous contact with cohorts of patients in the disciplines of internal medicine, neurology, obstetrics-gynecology, pediatrics, and psychiatry. With year long mentoring, students follow their patients through major venues of care. Surgery and radiology also are taught longitudinally, grounded in the clinical experiences of a cohort of patients and in a brief immersion experience working directly with an attending surgeon. Students participate in weekly, case-based tutorials integrating instruction in the basic sciences with training to address the common and important issues in medicine, as identified by national organizations. In addition, they participate in a social science curriculum that focuses on self-reflection, communication skills, ethics, population sciences, and cultural competence. In the pilot year (July 2004 to July 2005), HMS-CIC students performed at least as well as traditional students in tests of content knowledge and skills, as measured by National Board of Medical Examiners (NBME) Subject Exams and the fourth-year Objective Structured Clinical Exam, and they scored higher on a year-end comprehensive clinical skills self-assessment examination, suggesting that they retained content knowledge better. From surveys, HMS-CIC students were much more likely to see patients before diagnosis and after discharge and to receive feedback and mentoring from experienced faculty than were their traditionally educated peers. HMS-CIC students expressed more satisfaction with their curriculum and felt better prepared to cope with the professional challenges of patient care, such as being truly caring, involving patients in decision making, and understanding how the social context affects their patients. PMID- 17414199 TI - In-depth learning: one school's initiatives to foster integration of ethics, values, and the human dimensions of medicine. AB - Today's medical student curriculum is a lock-step experience that provides a broad survey of medicine with little opportunity to pursue fully integrated, in depth learning. To teach students about the human dimensions of health care, many schools simply have added courses that survey general areas such as ethics, values, and patient-doctor relationships. However, a superficial, broad-brush approach does not offer students sufficient opportunity to engage with these topics in substantive and meaningful ways. The authors propose that a theme based, individualized, in-depth learning experience (in which students pursue a focused project comprehensively and in detail)--one that is an integral part of the curriculum--helps students learn to blend values and ethics with medicine in a way that cannot occur during rapid-paced topical survey courses. Furthermore, it is in the depths of a learning experience that one comes face to face with the realities of uncertainty: the realization that unanswerable questions outnumber answerable ones; the awareness of the difficulty in accumulating sufficient evidence to answer a question that is, in fact, answerable; the recognition that many patients' problems transcend available evidence and must be addressed by the art of medicine; the realization that a patient can have a condition that one cannot diagnose and that may even get better for reasons that one cannot understand. The authors describe three initiatives at the University of Pittsburgh School of Medicine, two of which have been offered for more than 10 years, that illustrate the value of in-depth learning experiences. These in-depth experiences blend situated learning, reflective exercises, faculty mentoring, critical reading of literature, and constructive feedback in a prescribed but individualized curriculum. PMID- 17414200 TI - The new formal competency-based curriculum and informal curriculum at Indiana University School of Medicine: overview and five-year analysis. AB - There is growing recognition in the medical community that being a good doctor requires more than strong scientific knowledge and excellent clinical skills. Many key qualities are essential to providing comprehensive care, including the abilities to communicate effectively with patients and colleagues, act in a professional manner, cultivate an awareness of one's own values and prejudices, and provide care with an understanding of the cultural and spiritual dimensions of patients' lives. To ensure that Indiana University School of Medicine (IUSM) graduates demonstrate this range of abilities, IUSM has undertaken a substantial transformation of both its formal curriculum and learning environment (informal curriculum). The authors provide an overview of IUSM's two-part initiative to develop and implement a competency-based formal curriculum that requires students to demonstrate proficiency in nine core competencies and to create simultaneously an informal curriculum that models and supports the moral, professional, and humane values expressed in the formal curriculum. The authors describe the institutional and curricular transformations that have enabled and furthered the new IUSM curricular goals: changes in education administration; education implementation, assessment, and curricular design; admissions procedures; performance tracking; and the development of an electronic infrastructure to facilitate the expanded curriculum. The authors address the cost of reform and the results of two progress reviews. Specific case examples illustrate the interweaving of the formal competency curriculum through the students' four years of training, as well as techniques that are being used to positively influence the IUSM informal curriculum. PMID- 17414201 TI - Viewpoint: reflections on a well-traveled path: self-awareness, mindful practice, and relationship-centered care as foundations for medical education. AB - Medical students enter medical school hoping to have good relationships with their patients. Along with residents, however, they are exposed to a hidden curriculum that places the acquisition of biomedical knowledge above and at times at odds with development of the awareness and relationship skills important to the patient-physician relationship. Seasoned clinicians often enjoy the capacity for rich, healing relationships that are marked by reciprocal influence between them and their patients. The author argues that it is not necessary to relegate this recapturing of the human side of medicine to a midcareer epiphany, and the author calls for educational measures to encourage development of the communication and relationship-building skills throughout the medical education process. This will require a paradigm shift to a culture where teachers and learners are willing to consciously attend to their relationships and to work on self-awareness and mindfulness while they also master the biomedical knowledge required of the profession. Medical educators can facilitate and support continuous development of these skills throughout medical school and residency. Within the curriculum, there are many opportunities to teach how to reflect and to guide those reflections in ways that enhance our students' and residents' understanding of themselves as individuals and in the relationships they form with their patients. Using examples from narratives gathered in workshops and on work rounds with students and residents at the University of Washington School of Medicine, the author explores the concepts of relationship-centered care, self awareness, and mindfulness as proposed cornerstones of a new foundation for medical education. PMID- 17414202 TI - Medicine and the arts. Reveries of a solitary walker: [excerpt] by Jean-Jacques Rousseau. Commentary. PMID- 17414205 TI - Magnetic resonance imaging measurement of iron overload. AB - PURPOSE OF REVIEW: To highlight recent advances in magnetic resonance imaging estimation of somatic iron overload. This review will discuss the need and principles of magnetic resonance imaging-based iron measurements, the validation of liver and cardiac iron measurements, and the key institutional requirements for implementation. RECENT FINDINGS: Magnetic resonance imaging assessment of liver and cardiac iron has achieved critical levels of availability, utility, and validity to serve as the primary endpoint of clinical trials. Calibration curves for the magnetic resonance imaging parameters R2 and R2* (or their reciprocals, T2 and T2*) have been developed for the liver and the heart. Interscanner variability for these techniques has proven to be on the order of 5-7%. SUMMARY: Magnetic resonance imaging assessment of tissue iron is becoming increasingly important in the management of transfusional iron load because it is noninvasive, relatively widely available and offers a window into presymptomatic organ dysfunction. The techniques are highly reproducible within and across machines and have been chemically validated in the liver and the heart. These techniques will become the standard of care as industry begins to support the acquisition and postprocessing software. PMID- 17414206 TI - The pathophysiology, prevention, and treatment of stroke in sickle cell disease. AB - PURPOSE OF REVIEW: Stroke is one of the most devastating complications of sickle cell disease, but current research has led to improved understanding of its pathogenesis and to new approaches in the prevention of both primary and secondary stroke. This review focuses on advances reported in the past 2 years. RECENT FINDINGS: New concepts in the pathophysiology of central nervous system events in sickle cell disease have centered around hemolytic anemia and nitric oxide metabolism. Genetic risk factors are now being explored. Major improvement in primary stroke prevention has occurred through transcranial Doppler ultrasonography screening, but utilization of this technique is far from optimal. Hydroxyurea is now being tested as an alternative approach to chronic transfusion for secondary stroke prevention through a multicenter trial. Other studies are addressing the management of silent infarcts and nocturnal hypoxemia. SUMMARY: Increased understanding of the etiology and pathogenesis of stroke in sickle cell disease should eventually lead to improved management of all central nervous system complications. Alternative secondary stroke prevention with hydroxyurea may allow patients to avoid dependence on life-long chronic transfusion. Primary stroke prevention through transcranial Doppler ultrasonography screening may ultimately yield a dramatic reduction in the incidence of stroke in sickle cell disease. PMID- 17414207 TI - Spectrin-based skeleton in red blood cells and malaria. AB - PURPOSE OF REVIEW: Malaria represents one of the most important selective factors affecting human populations. Several inherited diseases of red blood cells lead to resistance at the erythrocytic stage. Among patients who experience hereditary elliptocytosis related to mutations of erythrocyte membrane proteins, molecular studies have shown the prevalence of particular spectrin mutations in patients from black ethnic extraction, leading one to question the selection of new malaria-resistant genes. RECENT FINDINGS: Prospective epidemiological and molecular studies in West Africa have confirmed the prevalence (between 0.6 and 1.6%) of particular spectrin mutations related to hereditary elliptocytosis. These studies have also revealed the frequency of alpha-spectrin chain polymorphisms, associated in cis with elliptocytogenic spectrin mutations and defining particular spectrin allele haplotypes. Culture studies of Plasmodium falciparum in elliptocytes bearing such elliptocytogenic alleles of spectrin showed that these alleles are supplementary genetic factors of malaria resistance in vitro. SUMMARY: Certain instances of spectrin mutations or polymorphisms have not yet been shown to constitute new factors of innate resistance to malaria in vivo. Epidemiological surveys of hereditary elliptocytosis and parasite culture studies, however, have argued that the relationships between parasite and spectrin-based skeleton should be examined more closely and the molecular interactions between parasite ligands and particular spectrin chain domains should be characterized. PMID- 17414208 TI - Erythrocyte remodeling by malaria parasites. AB - PURPOSE OF REVIEW: Plasmodium falciparum causes the most virulent form of human malarias. It is a protozoan parasite that infects human erythrocytes and the erythrocytic stages are responsible for all symptoms and pathologies of the disease. Critical to infection is the formation of a parasitophorous vacuolar membrane at the time of entry and within which the intracellular parasite proliferates. Since erythrocytes lack endocytic machinery, it is surprising that they can be infected by pathogens. This review summarizes recent studies of the erythrocyte-malaria interaction that have provided insights into properties of erythrocyte membranes as well as parasite mechanisms that remodel the erythrocyte. RECENT FINDINGS: Themes revealed by recent literature suggest that both parasite and erythrocyte components regulate parasite entry and intracellular growth by extensively remodeling host membranes. These remodeling events include the invagination of the host cell membrane during parasite entry that results in the creation and maintenance of a vacuole that surrounds the intracellular organism, and the development of antigenic, structural and transport alterations during intracellular parasite development. SUMMARY: The implications are that malarial erythrocyte remodeling events occur at a significant cost to the human host since many of the associated virulence events have been linked to severe disease pathologies. PMID- 17414209 TI - Prion protein and the red cell. AB - PURPOSE OF REVIEW: This review focuses on transfusion-transmission of variant Creutzfeldt-Jakob disease by red cell preparations. RECENT FINDINGS: Recently, three cases of probable transmission of variant Creutzfeldt-Jakob infectivity by transfusion of red cell preparations have been described in humans. Experiments on transmissible spongiform encephalopathies affecting rodents have led to the conclusion that infectivity in red cell preparations is not bound to the red cells themselves but contained within the suspending medium from which it can be removed by filtration. SUMMARY: Red cell preparations are the main transfusion product provided by blood services. If experiments demonstrating significant removal of rodent transmissible spongiform encephalopathy infections by filtration of red cell preparations are applicable to variant Creutzfeldt-Jakob in humans then a method for rendering human red cell preparations safe for transfusion is provided. PMID- 17414210 TI - Molecular insights into stress erythropoiesis. AB - PURPOSE OF REVIEW: In addition to its essential role in baseline erythropoiesis, the hormone erythropoietin drives the erythropoietic response to hypoxic stress. A mechanistic understanding of stress erythropoiesis would benefit multiple clinical settings, and may aid in understanding leukemogenesis. RECENT FINDINGS: The spectrum of progenitors targeted by the erythropoietin receptor is broader during stress than during baseline erythropoiesis. Further, the requirement for erythropoietin receptor signaling is more stringent during stress. However, erythropoietin receptor signaling has been mostly studied in vitro, where it is difficult to relate signaling events to stress-dependent changes in erythroid homeostasis. Here we review advances in flow cytometry that allow the identification and study of murine erythroid precursors in hematopoietic tissue as they are responding to stress in vivo. The death receptor Fas and its ligand, FasL, are coexpressed by early splenic erythroblasts, suppressing erythroblast survival and erythropoietic rate. During stress, erythropoietin receptor signaling downregulates erythroblast Fas and FasL, consequently increasing erythropoietic rate. SUMMARY: Erythropoietic rate is regulated at least in part through the erythropoietin receptor-mediated survival of splenic early erythroblasts. Future research will delineate how multiple antiapoptotic pathways, potentially activated by the erythropoietin receptor, interact to produce the remarkable dynamic range of erythropoiesis. PMID- 17414211 TI - Recombinant factor VIIa: its background, development and clinical use. AB - PURPOSE OF REVIEW: To examine the development of recombinant FVIIa (rFVIIa); a new concept of inducing hemostasis. It was developed for use in hemophilia patients with inhibitors against FVIII or FIX with the vision to provide these patients with a therapeutical option to be used instead of FVIII or FIX. For the first time it was shown that pharmacological doses of FVIIa induced hemostasis. RECENT FINDINGS: Hemostasis was achieved by rFVIIa in major surgery (repeated doses) as well as in a home-treatment setting (one single injection) in severe hemophilia patients with inhibitors. A recent study indicates that rFVIIa may be useful as prophylaxis. In heavily bleeding nonhemophilia patients rFVIIa was shown to induce hemostasis. Pharmacological doses of rFVIIa enhance thrombin generation on activated platelets resulting in the formation of tight hemostatic fibrin plugs resistant against premature proteolysis. High doses of rFVIIa seem to be safe probably due to its localized effect. SUMMARY: Pharmacological doses of rFVIIa induce hemostasis in severe hemophilia and in nonhemophilia patients with profuse, heavy bleeding. rFVIIa enhances thrombin generation on activated platelets, thereby initiating the formation of strong, tight fibrin hemostatic plugs resistant to premature lysis. It also seems to be safe in high doses. PMID- 17414212 TI - Protease-activated receptor signaling: new roles and regulatory mechanisms. AB - PURPOSE OF REVIEW: Protease-activated receptors are G-protein-coupled receptors that transmit cellular responses to coagulant proteases in a variety of cell types in the vasculature and other tissues. Several other proteases can activate protease-activated receptors in vitro and may affect their function in vivo. While a role for these receptors in hemostasis and thrombosis has been established, their functions in inflammatory and other responses have yet to be fully elucidated. In addition, the mechanisms responsible for protease and cell type-specific signaling mediated by these receptors are largely undefined. Here, we highlight recent advances in understanding the roles and regulation of protease-activated receptor signaling. RECENT FINDINGS: Recent studies have increased our knowledge of the function of protease-activated receptor signaling in platelets and its contribution to thrombosis. In other cell types, recent work has revealed new connections between these receptors and signaling effectors important for vascular development and inflammatory responses. Other studies have advanced our understanding of protease and cell type-specific responses as well as novel regulatory mechanisms for control of protease-activated receptor signaling. SUMMARY: Thus, elucidating the signaling and regulatory mechanisms of protease-activated receptors in various tissues and cell types is important for understanding their biological function as well as for designing therapeutic strategies to control their function. PMID- 17414213 TI - Fibrinogen and fibrin: scaffold proteins in hemostasis. AB - PURPOSE OF REVIEW: Elevated fibrinogen is a cardiovascular risk factor. Recent work provides a rationale for this risk, as abnormal fibrin clot structure, strength and stability correlates with coronary artery disease. This review describes in-vitro experiments whose intent is to define the molecular mechanisms that control clot architecture and function in vivo. RECENT FINDINGS: Biochemical and structural data continue to define the interactions between monomer units that assemble into a fibrin clot. In particular, 'A: a' interactions dominate the first step in fiber formation, while the analogous 'B: b' interactions have a minor role. Studies show the N-terminus of Bbeta, the C-terminus of Aalpha, and the splice variant gamma' modulate fibrin clot structure. Measurement of the mechanical properties of fibrinogen and fibrin show fibrin fibers are among the strongest in nature. Studies have identified fibrinogen-binding proteins that influence clot structure and function. SUMMARY: These findings defined mechanisms that control fibrin clot structure, strength and stability. This basic information provides direction for clinical studies to examine clot properties in pathologic thrombosis and pharmaceutical studies to develop therapeutic interventions to prevent or control cardiovascular disease. These studies also establish novel techniques to examine individual bonds, molecules and fibers. PMID- 17414214 TI - Fibrinolysis and the risk of venous and arterial thrombosis. AB - PURPOSE OF REVIEW: The fibrinolytic system is often regarded as just an innocent bystander in the pathogenesis of venous and arterial thrombosis, while (hyper)coagulation as a risk factor has been studied extensively. In this review, we evaluated studies that investigated the association between fibrinolysis and thrombosis. RECENT FINDINGS: There is some evidence for an association between impaired overall fibrinolytic activity and increased risk of venous or arterial thrombosis. Plasminogen levels were found not to be related to thrombosis. Plasma levels of tissue-type plasminogen activator were related to arterial thrombosis in a number of studies but not to venous thrombosis. Thrombin activatable fibrinolysis inhibitor levels appeared to be associated with venous thrombosis. Studies on the association between thrombin activatable fibrinolysis inhibitor or plasminogen activator inhibitor-1 and arterial thrombosis had conflicting results. SUMMARY: Current evidence on an association between fibrinolysis and thrombosis is inconclusive. Although overall assays point to an association, not all individual factors have an association with thrombosis. Most importantly, plasminogen deficiency is not related to thrombosis, which suggests that the fibrinolytic system as a whole is unimportant in the occurrence of thrombosis. Certain components of the fibrinolytic system, however, appear to be involved in processes unrelated to fibrin degradation but related to other processes important in the development of thrombosis. PMID- 17414215 TI - Calpain-mediated regulation of platelet signaling pathways. AB - PURPOSE OF REVIEW: There is considerable interest in understanding the function and mechanism of calpains in platelet aggregation, spreading, and granular secretion pathways. Recent insights from the calpain-1 knockout platelets suggest a pivotal role of these cysteine proteases in the regulation of outside-in signaling, aggregation, and clot retraction. RECENT FINDINGS: The calpain-1 knockout mouse provided direct evidence for the role of calpain-1 in platelet aggregation and clot retraction. Reduced tyrosine phosphorylation of platelet proteins correlated with reduced platelet aggregation and clot retraction. Future investigations of the mechanism of platelet defects in calpain-1 null mice may unveil the physiological functions of this important and elusive protease in mammalian cells. SUMMARY: This review focuses on the role of calpains in platelets with a particular emphasis on recent findings in calpain-1 null platelets. Previous studies used synthetic inhibitors to study the role of calpains in platelet function yielding useful information about the identification of calpain substrates. The development of calpain-1 null mice demonstrated that calpain-1 plays an important function in the regulation of platelet aggregation and clot retraction. Since the combined deletion of calpain 1 and calpain-2 genes results in embryonic lethality, the calpain-1 null mouse remains the only experimental model available to study the physiological role of calpains in mammalian cells. PMID- 17414216 TI - Tickling the tails: cytoplasmic domain proteins that regulate integrin alphaIIbbeta3 activation. AB - PURPOSE OF REVIEW: Integrin alphaIIbbeta3 activation is essential for platelet aggregation and related hemostatic events. In recent years, intense effort has been put forward to understand the molecular mechanisms regulating platelet integrin alphaIIbbeta3 activation. Here we review the current models of alphaIIbbeta3 activation and highlight the potential regulatory roles of proteins that interact directly with the alphaIIbbeta3 cytoplasmic domains, with emphasis on the alphaIIb cytoplasmic domain binding protein, CIB1. RECENT FINDINGS: Mutational and crystallographic studies reveal the importance of integrin transmembrane and cytoplasmic domains in propagating bidirectional signaling events. Proteins that interact directly with the integrin cytoplasmic domains may play important roles in mediating these signaling events. Of particular interest is the interaction between CIB1 and the alphaIIb tail which may function to negatively regulate alphaIIbbeta3 activation. In addition, a number of CIB1 interacting proteins have been identified, including p21-activated kinase and serum-inducible kinase, which may act in concert with CIB1 to regulate platelet function. SUMMARY: Understanding the molecular mechanisms underlying integrin activation will be important in developing novel therapies to regulate platelet function in cardiovascular disease. Discussion of recent developments in elucidating the mechanism of integrin activation, with particular focus on the platelet integrin alphaIIbbeta3, is provided in this review. PMID- 17414217 TI - Signaling and regulation of the platelet glycoprotein Ib-IX-V complex. AB - PURPOSE OF REVIEW: The platelet adhesion receptor, the glycoprotein Ib-IX-V complex, not only mediates platelet adhesion but also transmits signals leading to platelet activation, aggregation and secretion. Significant progress has been made recently on the signaling pathways and regulatory mechanisms involving glycoprotein Ib-IX-V function. RECENT FINDINGS: The interaction of glycoprotein Ib-IX-V with its ligand, von Willebrand factor, is dually controlled by von Willebrand factor conformation and intracellular signal-mediated regulation of glycoprotein Ib-IX-V receptor function that requires the zeta isoform of the 14-3 3 protein family (14-3-3zeta). Glycoprotein Ib-IX-V signaling is mediated by the Src family of protein kinases, phospholipase C, calcium elevation, phosphoinositol 3-kinase, and multiple amplification mechanisms including the nitric oxide-cGMP pathway, the mitogen-activated protein kinase pathway, the immunoreceptor tyrosine-based activation motif pathway, and ADP and thromboxane A2 pathways. SUMMARY: Progress in understanding the mechanism(s) regulating glycoprotein Ib-IX-V should help develop inhibitors and modifiers that interfere or augment its von Willebrand factor binding function and thus be useful for treating thrombosis and bleeding disorders. Characterization of intracellular molecules and pathways in glycoprotein Ib-IX-V signaling has implications in the development of new agents and for the use of existing drugs that affect glycoprotein Ib-IX-V signaling. PMID- 17414218 TI - Structural and functional correlation of ADAMTS13. AB - PURPOSE OF REVIEW: ADAMTS13 represents a landmark in a journey that began over 80 years ago with a single clinical case. Thrombotic thrombocytopenic purpura exemplifies how von Willebrand factor can be responsible for life-threatening thrombosis. This review summarizes recent progress on ADAMTS13, which prevents this deadly event. RECENT FINDINGS: Recent advances are summarized in four main areas. First, the core ADAMTS13-binding site is contained in a short sequence in the A2 domain, but other domains affect this interaction. Mutations from thrombotic thrombocytopenic purpura and von Willebrand disease provide clues for the structural prerequisites and regulation of von Willebrand factor cleavage. Second, studies are unraveling the reasons why urea, BaCl2, and low ionic strength are required to cleave von Willebrand factor under static conditions. Third, studies on thrombotic thrombocytopenic purpura and ADAMTS13-knockout mice suggest that ADAMTS13 deficiency alone may not be sufficient to cause thrombotic thrombocytopenic purpura. Finally, ADAMTS13 could be an antithrombotic agent for thrombotic thrombocytopenic purpura and other thrombotic conditions. SUMMARY: Study of ADAMTS13 has exploded since this metalloprotease was characterized. This knowledge reveals the nature of ADAMTS13's interaction with von Willebrand factor and the pathogenesis of clinical thrombotic thrombocytopenic purpura, especially in relation to ADAMTS13. PMID- 17414219 TI - ADAMTS13 assays and ADAMTS13-deficient mice. AB - PURPOSE OF REVIEW: Thrombotic thrombocytopenic purpura can be induced by acquired or congenital deficiency of the plasma von Willebrand factor-cleaving protease, ADAMTS13. Measurement of ADAMTS13 activity is important for the diagnosis and treatment of microangiopathies including thrombotic thrombocytopenic purpura. Phenotypic analysis of mice lacking the Adamts13 gene is valuable for understanding the pathogenesis of microangiopathies. RECENT FINDINGS: The minimum substrate for ADAMTS13 activity was identified as 73 amino acid residues in the A2 domain of von Willebrand factor, called VWF73. Several new assays have been developed using this sequence. The VWF73-based assays are rapid, quantitative, and easy to handle, and are well correlated with the measures from previous assays. Mice lacking the Adamts13 gene were produced. The mice were viable and fertile. They showed a prothrombotic state but no symptoms of spontaneous thrombocytopenia, hemolytic anemia, or microvascular thrombosis were observed. SUMMARY: VWF73-based ADAMTS13 assays will significantly facilitate the accurate diagnosis of microangiopathies and contribute to the improved clinical treatment of these diseases. Accumulated clinical information on patients with ADAMTS13 deficiency and mice lacking the Adamts13 gene indicates that additional environmental or genetic susceptibility factors are required to trigger thrombotic thrombocytopenic purpura. PMID- 17414220 TI - The presence of active von Willebrand factor under various pathological conditions. AB - PURPOSE OF REVIEW: To highlight mechanisms that regulate the balance between latent and active von Willebrand factor (VWF), and describe pathological conditions leading to increased levels of active VWF. RECENT FINDINGS: Levels of circulating active VWF are increased in von Willebrand disease type 2B, HELLP syndrome, malaria and antiphospholipid syndrome. SUMMARY: Freshly secreted VWF consists of ultra-large multimers that interact spontaneously with platelets at the endothelial cell surface. Proteolysis of ultra-large VWF by a member of the disintegrin and metalloprotease with thrombospondin motif family (ADAMTS13) reduces both multimeric size and accessibility of platelet-adhesion sites. The resulting VWF molecules circulate as inactive multimers, which regain their platelet-adhesion capacity upon binding to the subendothelial matrix, in particular under conditions of high shear. Unfortunately, mechanisms responsible for suppression of circulating plasma levels of active VWF are hampered in a number of pathological conditions, leading to VWF-platelet aggregates associated with thrombotic complications or thrombocytopenia. A recently developed assay allowed us to monitor the presence of circulating active VWF and we found that several diseases are characterized by increased levels. Further analysis provided insight into mechanisms contributing to the presence of active VWF, which revealed that beta2-glycoprotein I may act as a natural regulator of VWF-platelet interactions. PMID- 17414221 TI - Bibliography. Current world literature. Erythroid system and its diseases. PMID- 17414222 TI - Anti-ischemic effects of fasudil, a specific Rho-kinase inhibitor, in patients with stable effort angina. AB - Epicardial coronary stenosis causes myocardial ischemia; however, the role of coronary microvessels is poorly understood in the pathogenesis of effort angina. We have previously demonstrated that Rho-kinase pathway is substantially involved in coronary arterial hyperconstriction in patients with vasospastic angina and those with microvascular angina. In the present study, we tested our hypothesis that Rho-kinase is involved in coronary microvascular constriction in patients with effort angina. Intracoronary administration of fasudil (300 microg/min for 15 min), a specific Rho-kinase inhibitor, significantly increased oxygen saturation in coronary sinus vein from 37 +/- 3% to 41 +/- 3% (P < 0.05) but not in six age-matched controls (from 42 +/- 3% to 43 +/- 3%, P = NS). Furthermore, the fasudil treatment significantly ameliorated pacing-induced myocardial ischemia in patients with effort angina (magnitudes of symptom: 1.5 +/- 0.6 to 0.6 +/- 0.4, P < 0.01; ischemic ST-segment depression, 1.8 +/- 0.3 to 1.0 +/- 0.2 mm, P < 0.01; percent lactate production, 50 +/- 17% to 0.4 +/- 7%, P < 0.01) without significant hemodynamic changes. These results provide the first evidence that Rho-kinase is substantially involved in coronary microvascular dysfunction associated with myocardial ischemia in patients with effort angina, suggesting that Rho-kinase can be a novel therapeutic target in ischemic heart disease. PMID- 17414223 TI - Pitavastatin restores vascular dysfunction in insulin-resistant state by inhibiting NAD(P)H oxidase activity and uncoupled endothelial nitric oxide synthase-dependent superoxide production. AB - 3-Hydroxyl-3-methylglutaryl coenzyme A reductase inhibitors (statins) may benefit the vasculopathy of insulin resistance independent of its lipid-lowering effects. Because imbalance of nitric oxide (NO) and superoxide anion (O(2)(-)) formation may lead to vascular dysfunction, we investigated the effect of statin on vasomotion of insulin-resistant state to clarify the mechanism by which statin ameliorates the impaired function. In the isolated aorta, contraction induced by angiotensin II was more potent in Zucker fatty rats (ZF) compared with that in Zucker lean rats. Both angiotensin II type 1 receptor expression and O(2)(-) production were upregulated in ZF. In addition, deficiency of tetrahydrobiopterin (BH4) contributes to the endothelial dysfunction in ZF. Oral administration of pitavastatin for 8 weeks normalized angiotensin II-induced vasoconstriction and endothelial function in ZF. Pitavastatin treatment of ZF increased vascular BH4 content, which was associated with twofold increase in endothelial NO synthase (eNOS) activity as well as a 60% reduction in endothelial O(2)(-) production. The treatment also markedly downregulated protein expression of angiotensin II type 1 receptor and gp91phox, whereas expression of guanosine triphosphate cyclohydrolase I was upregulated. Pitavastatin restores vascular dysfunction by inhibiting NAD(P)H oxidase activity and uncoupled eNOS-dependent O(2)(-) production. PMID- 17414224 TI - Oral administration of both tetrahydrobiopterin and L-arginine prevents endothelial dysfunction in rats with chronic renal failure. AB - We examined the mechanism of endothelial dysfunction in chronic renal failure (CRF), with reference to NO synthase. CRF was induced by 5/6 nephrectomy in rats. Either L-arginine (1.25 g/L in drinking water), tetrahydrobiopterin (BH4, 10 mg/kg per day in food), or a combination of the 2 were orally administered to CRF rats for 9 weeks. CRF rats showed elevation of systolic blood pressure compared with sham-operated rats. Endothelium-dependent relaxation induced by acetylcholine or A23187 in the isolated aorta was significantly reduced, and in vitro treatment with L-arginine, BH4, or superoxide dismutase restored the relaxation. Aortic segments from CRF rats showed significantly higher superoxide production in response to A23187, which was inhibited by L-NAME. Plasma concentrations of asymmetric dimethylarginine and symmetric dimethylarginine were higher in CRF rats. These changes in CRF rats were totally or partially decreased by L-arginine or BH4 supplementation in vivo. Interestingly, the combined treatment showed additive effects in certain parameters. These results suggest that vascular disorders in CRF rats may be partly due to NOS uncoupling caused by a relative deficiency of BH4 and partially due to accumulation of endogenous inhibitors of NOS and L-arginine uptake, resulting in the decrease of NO production and the increase of reactive oxygen species. PMID- 17414225 TI - Anti-atherogenic effects of a mixture of ascorbic acid, lysine, proline, arginine, cysteine, and green tea phenolics in human aortic smooth muscle cells. AB - Certain drastic behavioral modifications by arterial wall smooth muscle cells (SMC) have been considered key steps in the formation of atherosclerotic lesions: massive migration of SMC from the media to the intima layer of the vessel, dedifferentiation of SMC to proliferating phenotype, and increased secretion of inflammatory cytokines as a response to inflammatory stimuli. We investigated the anti-atherogenic effects of naturally occurring compounds (ascorbic acid, green tea extract, lysine, proline, arginine, and N-acetyl cysteine) using the model of cultured aortic SMC. Cell growth was measured by DNA synthesis, cell invasiveness was measured through Matrigel, matrix metalloproteinase-2 (MMP-2) secretion was measured by zymography, and SMC secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) was measured by immunochemistry. Fetal bovine serum-stimulated SMC growth was inhibited by the nutrient mixture (NM) with 85% inhibition at 100 microg/mL. A corresponding concentration of epigallocatechin gallate (EGCG; 15 microM), the most active tea phenolic, produced a significant effect but one lower than NM. NM inhibited aortic SMC Matrigel invasion in a dose dependent manner and significantly decreased MMP-2 expression. Stimulation of SMC with tumor necrosis factor-alpha significantly increased production and secretion of such mediators of inflammation as IL-6 and MCP-1; addition of 100 microg/mL NM inhibited secretion of MCP-1 and IL-6 by 65% and 47%, respectively. These data suggest that the NM of ascorbic acid, tea phenolics, and selected amino acids has potential in blocking the development of atherosclerotic lesions by inhibiting atherogenic responses of vascular SMC to pathologic stimuli and warrants in vivo studies. PMID- 17414226 TI - QT effects of duloxetine at supratherapeutic doses: a placebo and positive controlled study. AB - BACKGROUND: The electrophysiological effects of duloxetine at supratherapeutic exposures were evaluated to ensure compliance with regulatory criteria and to assess the QT prolongation potential. METHODS: Electrocardiograms were collected in a multicenter, double-blind, randomized, placebo-controlled, crossover study that enrolled 117 healthy female subjects aged 19 to 74 years. Duloxetine dosages escalated from 60 mg twice daily to 200 mg twice daily; a single moxifloxacin 400 mg dose was used as a positive control. Data were analyzed using 3 QT interval correction methods: mixed-effect analysis of covariance model with RR interval change from baseline as the covariate, the QT Fridericia's correction method, and the individual QT correction method. Concentrations of duloxetine and its 2 major metabolites were measured. RESULTS: Compared with placebo, the mean change from baseline in QTc decreased with duloxetine 200 mg twice daily. The upper limits of the 2-sided 90% confidence intervals for duloxetine vs. placebo were <0 msec at each time point by any correction method. No subject had absolute QT Fridericia's correction values >445 msec with duloxetine, and the change in QT Fridericia's correction from baseline with duloxetine did not exceed 36 msec. No relationship was detected between QTc change and plasma concentrations of duloxetine or its metabolites even though average duloxetine concentrations ranged to more than 5 times those achieved at therapeutic doses. Moxifloxacin significantly prolonged QTc at all time points, regardless of correction method. CONCLUSIONS: Duloxetine does not affect ventricular repolarization as assessed by both mean changes and outliers in QT corrected by any method. PMID- 17414227 TI - Role of Na+/H+ exchanger in the pathogenesis of ischemic acute renal failure in mice. AB - We evaluated the effects of 5-(N-ethyl-N-isopropyl) amiloride (EIPA), a Na+/H+ exchanger (NHE) inhibitor, on ischemia/reperfusion (I/R)-induced acute renal failure (ARF) in mice. Ischemic ARF was induced by clamping the left renal artery and vein for 40 minutes followed by reperfusion 2 weeks after the contralateral nephrectomy. Preischemic treatment with EIPA attenuated the I/R-induced renal dysfunction. Histopathological examination of the kidney of ARF mice revealed severe renal damage such as tubular necrosis and proteinaceous casts in the tubuli. Histologically evident damage was also improved by preischemic treatment with EIPA. In addition, the I/R-induced increase in renal endothelin-1 (ET-1) content was suppressed by preischemic treatment with EIPA, reflecting the difference in immunohistochemical ET-1 localization in necrotic tubular epithelium. However, the postischemic treatment with EIPA failed to improve the I/R-induced renal dysfunction and ET-1 overproduction. These findings suggest that NHE activation, followed by renal ET-1 overproduction, plays an important role in the pathogenesis of I/R-induced renal injury. The inhibition of NHE by EIPA may be considered as a therapeutic approach to protect the postischemic ARF. PMID- 17414228 TI - Add-on amlodipine improves arterial function and structure in hypertensive patients treated with an angiotensin receptor blocker. AB - The present study was designed to determine whether adding amlodipine further improved functional and structural cardiovascular damage in hypertensive patients whose blood pressure was already well controlled with an angiotensin II type 1 receptor blocker (ARB). The cardiothoracic ratio on chest radiographs, level of urinary albumin excretion, pulse wave velocity (PWV), intima-media thickness (IMT) of the carotid arteries, and 24 hour ambulatory blood pressure (BP) were evaluated before and 12 months after the start of add-on of amlodipine or placebo in 50 hypertensive patients being treated with an ARB. The add-on amlodipine therapy significantly improved the PWV from 1689 +/- 61 to 1447 +/- 47 cm/s and the IMT from 0.88 +/- 0.08 to 0.75 +/- 0.06 mm in the hypertensive patients treated with an ARB without altering their mean 24 hour ambulatory BP values, but did not alter the cardiothoracic ratio or urinary albumin excretion. Amlodipine also significantly decreased the variability of ambulatory BP, but the decrease did not significantly contribute to the changes in PWV or IMT. Thus, the add-on low-dose amlodipine therapy had benefits in terms of the vascular function and vascular structure of hypertensive patients treated with an ARB that were independent of its depressor effects. The antiatherogenic pleiotropic properties of amlodipine have a preventive effect on the progression of arterial stiffness in hypertensive patients treated with an ARB. PMID- 17414229 TI - Dose-dependent inhibition of human platelet aggregation by prasugrel and its interaction with aspirin in healthy subjects. AB - The aims of this open-label, randomized, dose-escalation pharmacodynamic study of prasugrel, an orally active antiplatelet agent, were to assess its interaction with aspirin (ASA, 325 mg) in healthy subjects after a loading dose (LD) and subsequent 5 days of once-daily maintenance doses (MD) of prasugrel or the active comparator, clopidogrel. We measured platelet aggregation induced by ADP, collagen, and TRAP and compared effects on maximal and residual platelet aggregation responses. On a background of ASA, subjects were randomly assigned to 1 of 4 prasugrel treatment groups (LD/MD in mg: 20/5, 30/7.5, 40/10, or 60/15; n = 8/group) or to clopidogrel 300 mg LD/75 mg MD (n = 11). Prasugrel dose dependently inhibited ADP-induced platelet aggregation and exhibited higher levels of platelet inhibition than clopidogrel or ASA alone. Prasugrel plus ASA resulted in additive inhibition of collagen- and TRAP-induced platelet aggregation. Although inhibition of residual aggregation was greater than inhibition of maximal aggregation, values were highly correlated. The safety and tolerability of prasugrel plus ASA were also monitored. Within the limitations of the study, prasugrel was found to be well tolerated when dosed as LD followed by MD in the presence of ASA and provided greater platelet inhibition than ASA alone. PMID- 17414230 TI - Intermittent clonidine regimen abolishes tolerance to its antihypertensive effect: a spectral study. AB - The development of tolerance to the antihypertensive effect of clonidine and related imidazolines is clinically recognized. Here, we employed a restricted daytime (8:30 AM until 4:30 PM) clonidine regimen to establish a model of sustained hypotension in spontaneously hypertensive rats (SHRs). Blood pressure (BP), heart rate (HR), and myocardial contractility (dP/dt(max)) were measured by radiotelemetry in pair-fed SHRs receiving liquid diets with or without clonidine (150 microg/kg per day) for 12 weeks. The cardiovascular autonomic control was assessed by power spectral analysis [fast Fourier transformations (FFT)] of hemodynamic variability. Clonidine had no effect on dP/dt(max) and significantly decreased BP and HR during the 8 hour exposure periods throughout the study duration. BP returned to control levels during overnight periods, with no signs of rebound hypertension. FFT analysis of interbeat intervals (IBI) showed pronounced decreases and increases of spectral powers in low-frequency (IBI-LF, 0.20-0.75 Hz) and high-frequency (IBI-HF, 0.75-3 Hz) bands, respectively, in clonidine-treated rats. The IBI(LF/HF) ratio was significantly reduced by clonidine, suggesting cardiac parasympathetic dominance. Clonidine also decreased the vasomotor sympathetic tone, as reflected by the reduced BP-LF spectral density. The sympathoinhibitory effect of clonidine is further confirmed by the significant reductions in urinary norepinephrine levels. Clonidine increased urine output during the 8 hour treatment period but not during the 24 hour period. Plasma and urine osmolality and electrolytes were not altered by clonidine. It is concluded that by adopting the limited-access paradigm, tolerance to the hypotensive and sympathoinhibitory actions of clonidine and, possibly, its side effects, could be minimized. PMID- 17414231 TI - An open-label trial of aripiprazole augmentation for treatment-resistant generalized anxiety disorder. PMID- 17414232 TI - Olanzapine-associated bilateral eyelid edema. PMID- 17414233 TI - Is it easier to find what you are looking for if you think you know what it looks like? PMID- 17414234 TI - Outcomes for Latin American versus White patients suffering from acute mania in a randomized, double-blind trial comparing olanzapine and haloperidol. AB - Data from a published double-blind randomized trial comparing olanzapine versus haloperidol in acute mania were used to address the response and tolerability of Latin American patients. Primary efficacy end point was the remission rate (Young Mania Rating Scale score 60 ng/mL, DDS 7.9), and all concentrations of fluoxetine (P < 0.05, DDS 7.1). Significant improvement in everyday function mirrored findings for pain intensity. CONCLUSIONS: Preliminary evidence for a low concentration "therapeutic window" for noradrenergic analgesia may warrant additional study. PMID- 17414236 TI - Bupropion in the treatment of pathological gambling: a randomized, double-blind, placebo-controlled, flexible-dose study. AB - We tested the efficacy of bupropion in the treatment of persons with pathological gambling (PG). Nondepressed, healthy subjects with Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition PG were randomly assigned to placebo or flexibly dosed bupropion in a 12-week double-blind trial. Outcome measures included the Yale-Brown Obsessive-Compulsive Scale modified for PG, the Gambling Severity Assessment Scale, the Clinical Global Impression Improvement and Severity Scales, the Global Assessment Scale, the Timeline Follow Back, the Attention-Deficit/Hyperactivity Disorder Rating Scale, and the Sheehan Disability Scale. Thirty-nine subjects (28 men, 11 women) were randomized to bupropion (n = 18) or placebo (n = 21). The 2 groups were similar on demographic and clinical measures. There were few differences between the treatment groups on any primary or secondary outcome measure, although subjects in each cell experienced significant improvement. Of subjects with at least 1 postrandomization visit, 35.7% of bupropion and 47.1% of placebo recipients experienced "much" or "very much" improvement on the Clinical Global Impression Improvement Scale. The trial was complicated by a high noncompletion rate (43.6%). Bupropion was well tolerated. Bupropion and placebo recipients did equally well in a short-term trial, with improvement seen as early as the first week of treatment. The high placebo response rate and the high noncompletion rate each reflect the challenge inherent in treating persons with PG. PMID- 17414237 TI - Postintervention effect of nicotine replacement therapy for smoking reduction: a randomized trial with a 5-year follow-up. AB - We tested whether a reduction of cigarette consumption obtained after 6 months of nicotine replacement therapy was maintained 5 years after the end of this treatment. Heavy smokers (mean = 30 cigarettes per day) who had no intention of quitting smoking were randomly assigned to a 6-month treatment of nicotine (15-mg patch, 4-mg gum, and/or 10-mg inhaler, n = 265), placebo (n = 269), or no intervention (n = 389). Products were sent by mail, and education was limited to a booklet. Of 923 participants, 879 (95%) were followed after 6 months and 671 (73%) after 5 years. After 6 months, smoking reduction was larger for nicotine ( 10.9 cigarettes per day) than for placebo (-8.7) and no treatment (-4.9, all P or= 0.2) and smoking cessation rates (17% to 21%, all P > 0.2) were similar in all groups. In smokers, 5-year continuous abstinence was higher in those who had reduced their cigarette consumption by at least 50% between baseline and 6 months than in those who did not reduce (11.9% vs 5.6%; P = 0.011; odds ratio, 2.3; 95% confidence interval, 1.2-4.2). Thus, the initial effect of the treatment on smoking reduction was not maintained after 5 years. However, reducing cigarette consumption was associated with a higher chance of subsequently quitting smoking. Nicotine replacement therapy in unmotivated smokers had no deleterious effect on dependence levels and smoking behavior. PMID- 17414238 TI - Motor cortex excitability after vagus nerve stimulation in major depression. AB - Recent data suggest that inhibitory pathways may be involved in the pathophysiology of depression and in the mode of action of some antidepressant interventions. The aim of the present study was to test whether vagus nerve stimulation (VNS) can affect motor cortex excitability. Measures of motor cortical excitability were probed by using single-pulse and paired-pulse transcranial magnetic stimulation at baseline, after 10 weeks of left VNS, and additionally, in an on-off paradigm in 10 patients with treatment-resistant unipolar depression. Ten weeks of VNS was associated with a selective and pronounced increase in intracortical inhibition, whereas no changes occurred in the on-off paradigm. These results suggest that VNS is capable of changing motor cortical excitability in patients with depression. PMID- 17414239 TI - Naltrexone and disulfiram in patients with alcohol dependence and current depression. AB - OBJECTIVE: Although disulfiram and naltrexone have been approved by the Food and Drug Administration for the treatment of alcoholism, no medications have been approved for individuals with alcohol dependence and comorbid psychiatric disorders. In particular, the effect of these medications on alcohol use outcomes and on specific psychiatric symptoms is still unknown in patients with the most common co-occurring disorder, major depression. METHOD: Two hundred fifty-four patients with a major Axis I psychiatric disorder and comorbid alcohol dependence were treated for 12 weeks in an outpatient medication study conducted at 3 Veterans Administration outpatient clinics. Randomization included (1) open randomization to disulfiram or no disulfiram, and (2) double-blind randomization to naltrexone or placebo. This resulted in 4 groups: (1) naltrexone alone, (2) placebo alone, (3) disulfiram and naltrexone, and (4) disulfiram and placebo. Primary outcomes were measures of alcohol use. Secondary outcomes included psychiatric symptoms assessed by the Hamilton Depression Rating Scale, alcohol craving, gamma-glutamyltransferase levels, and adverse events. RESULTS: One hundred thirty-nine subjects (54.7%) met the current Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for major depression. There was no relationship between the diagnosis of depression and medication treatment on alcohol use outcomes, psychiatric symptoms, or the reporting of side effects for these medications. There was a significant interaction between diagnosis, medication group, and craving, where subjects with depression on disulfram reported lower craving over time than subjects with depression on naltrexone. CONCLUSIONS: The results suggest that disulfiram and naltrexone are safe pharmacotherapeutic agents for dually diagnosed individuals with depression for the treatment of alcohol use disorders. PMID- 17414240 TI - Failed efficacy of fluoxetine in the treatment of posttraumatic stress disorder: results of a fixed-dose, placebo-controlled study. AB - A multicenter, double-blind, 12-week, placebo-controlled trial of 411 randomized patients, predominantly women diagnosed with posttraumatic stress disorder, failed to show a difference between either dose of fluoxetine treatment and placebo. The mean changes from baseline (SD) measured by the Clinician Administered PTSD Scale scores were -42.9 (23.1), -42.8 (27.9), and -36.6 (25.7) in the 20-mg fluoxetine, 40-mg fluoxetine, and placebo arms, respectively. Placebo response rate was substantially higher in this study than in a previously published fluoxetine trial of posttraumatic stress disorder. PMID- 17414241 TI - Management of acute agitation in patients with bipolar disorder: efficacy and safety of intramuscular aripiprazole. AB - To investigate the efficacy and safety of intramuscular (IM) aripiprazole for the treatment of agitation in patients with bipolar I disorder, manic or mixed episodes. In total, 301 patients experiencing acute agitation were randomized to IM aripiprazole 9.75 mg per injection (n = 78), IM aripiprazole 15 mg per injection (n = 78), IM lorazepam 2 mg per injection (n = 70), or IM placebo (n = 75) in this double-blind multicenter study. Patients could receive up to 3 injections over 24 hours. Primary efficacy measure was mean change in Positive and Negative Syndrome Scale Excited Component score from baseline at 2 hours after first injection. Mean improvements in Positive and Negative Syndrome Scale Excited Component score at 2 hours were significantly greater with IM aripiprazole (9.75 mg, -8.7; 15 mg, -8.7) and IM lorazepam (-9.6) versus IM placebo (-5.8; P 20% in males; >10% in females) in these populations. Associations between TBI and disorders meeting Diagnostic and Statistical Manual of Mental Disorder, Fourth Edition criteria were strong in the SW community (adjusted odds ratio [OR]=2.4; 95% confidence interval [CI]=1.5 3.8), less so in the NP community (adjusted OR=1.4; 95% CI=0.9-2.2). CONCLUSIONS: TBI is prevalent among AIs and is associated with increased OR of neuropsychiatric difficulties. This association may vary between tribes, though no interaction effect was found. AIs with TBI may require more thorough psychiatric screening to promote healthier outcomes. PMID- 17414313 TI - Ethnic differences in rehabilitation placement and outcome after TBI. AB - OBJECTIVE: To determine whether race/ethnicity and proficiency with the English language influence access to rehabilitation services, and ultimately outcome after traumatic brain injury (TBI). DESIGN: A retrospective correlational investigation. SETTING AND PARTICIPANTS: Postrehabilitation outpatients with blunt TBI. A total of 476 patients were examined 6 months postinjury, of which 109 (23%) were Hispanics, and of those 42 were predominantly Spanish speaking. MAIN OUTCOME MEASURES: Access to rehabilitation services, Glasgow Outcome Scale Extended. RESULTS: Insurance, age, and injury severity had the greatest influence on receipt of rehabilitation services; however, higher rates of severe disability were found among Hispanics and Spanish speakers than non-Hispanic whites and non Hispanic English speakers, respectively. CONCLUSIONS: Insurance status has a larger influence on receipt of rehabilitation services than ethnicity or proficiency with the English language, but language proficiency is a significant predictor of severe disability. PMID- 17414314 TI - Racial differences in caregiving patterns, caregiver emotional function, and sources of emotional support following traumatic brain injury. AB - OBJECTIVE: Compare white and African American caregivers of people with moderate to severe traumatic brain injury (TBI) regarding caregiving patterns, emotional function and life satisfaction, and preferred supports. DESIGN: Prospective, observational study; 1, 2, or 5 years post-TBI. SETTING: Six TBI model systems. PARTICIPANTS: Two hundred fifty-six caregivers (195 white and 61 African American). MEASURES: Brief Symptom Inventory-18, Satisfaction With Life Scale. RESULTS: Races differed as to kinship patterns, with more white caregivers including spouses and more African Americans including "other relatives." African Americans spent significantly more time in direct caregiving, and reported more depression. African American TBI survivors were significantly more disabled than whites, which appeared to account for emotional function differences. Whites were more likely to use professional services for emotional support. CONCLUSION: Across races, TBI caregiver emotional health is affected by the functional level of the survivor. African American caregivers may be at risk for worse emotional consequences due to worse survivor outcomes, yet may underutilize professional services. PMID- 17414315 TI - The influence of cultural background on motivation for and participation in rehabilitation and outcome following traumatic brain injury. AB - AIMS: To compare motivation for and participation in rehabilitation, outcome, and distress over role changes in persons with traumatic brain injury (TBI) from the dominant English-speaking culture in Australia versus those from minority culturally and linguistically diverse (CALD) backgrounds. MAIN MEASURES: Motivation for Traumatic Brain Injury Rehabilitation Questionnaire; Craig Handicap Assessment Reporting Technique. PARTICIPANTS: Two groups of persons with TBI, 38 of English-speaking backgrounds and 32 of CALD backgrounds. RESULTS: Groups had similar education and preinjury employment status, both showed positive attitudes toward rehabilitation and participated equally in rehabilitation. However, CALD participants showed poorer outcomes in several domains, including postinjury employment status, cognitive independence, mobility and social integration, and showed greater distress about changes in ability to perform certain life roles. CONCLUSIONS: Differences in outcome and levels of distress over role changes may occur in those from CALD backgrounds following TBI, independent of socioeconomic background and access to rehabilitation. There is a need to further investigate possible reasons for this, including beliefs, coping style, and emotional response to injury. PMID- 17414316 TI - The new B7s: playing a pivotal role in tumor immunity. AB - B7-H1, B7-DC, B7-H2, B7-H3, and B7-H4, all new additions to the B7 family, here termed "the new B7s," are emerging as important tools in directing immune function; each with unique, yet often overlapping functions. Clearly, each B7 molecule has developed its own indispensable niche in the immune system. The expression of both stimulatory and inhibitory B7 molecules seems to play an essential role in regulating the immune response to transformed cells through a variety of mechanisms. As specific niches of B7 family members continue to be dissected, their diagnostic and therapeutic potential becomes ever more apparent. In this review, we will discuss the role of the new B7s in activation and inhibition of antitumor immune responses, their prospects in diagnostics, and also potential and developing immunotherapy protocols. PMID- 17414317 TI - Cellular and functional characterization of immunoresistant human glioma cell clones selected with alloreactive cytotoxic T lymphocytes reveals their up regulated synthesis of biologically active TGF-beta. AB - Two immunoresistant (IR) glioma cell variants, 13-06-IR29 and 13-06-IR30, were cloned from 13-06-MG glioma cell populations after receiving continuous immunoselective pressure from multiple alloreactive cytotoxic T lymphocyte (aCTL) preparations. Reapplication of aCTL immunoselective pressure to the IR clones, displaying a partial regain in sensitivity to aCTL after removal of the selective pressure, restored the resistance. The IR variants exhibited cross-resistance to non-human leukocyte antigen (HLA)-restricted effector cells and gamma irradiation, but not to carmustine. The IR clones were characterized for factors that might contribute to the immunoresistance. The aCTL adhesion to extracellular matrix extracts derived from either the IR clones or the parental cells was similar and not impaired. Furthermore, aCTL binding to parental cells and IR clones was equal. Down-regulation of the cell recognition molecules, class I HLA or intercellular adhesion molecule-1 (ICAM-1), that would inhibit their recognition by aCTL was not observed on the IR clones. The down-regulation of Fas by the IR clones correlated with their resistance to FasL-induced apoptosis. HLA G or FasL that might provide an immunotolerant environment or provide a means of counterattack to aCTL, respectively, were not associated with the IR phenotype. The aCTL, coincubated with the IR clones and parental cells, displayed up regulation of multiple secreted cytokines. A significant up-regulation of bioactive transforming growth factor (TGF)-beta was observed in the IR clones compared with the parental cells. These data suggest that increased secretion of bioactive TGF-beta may inhibit aCTL lysis of the IR clones. Disruption of the TGF beta signaling pathway may circumvent the resistance. PMID- 17414318 TI - New peptide vaccine candidates for epithelial cancer patients with HLA-A3 supertype alleles. AB - We previously identified 2 cancer-associated antigens, immediate early response gene X-1 (IEX) and small GTPase (Ran), and their 5 epitopes using human leukocyte antigen (HLA)-A33-restricted and tumor-infiltrating T cells from a colon cancer patient. In this study, we examined whether or not these peptides can induce cytotoxic T lymphocytes (CTLs) in HLA-A11+ or HLA-A31+ epithelial cancer patients because the HLA-A11, HLA-A31, and HLA-A33 alleles share binding motifs as an HLA A3 supertype family, which is widely distributed in many ethnic populations. Among them, the 2 peptides, IEX 47-56 and IEX 61-69, induced peptide-specific CTLs from peripheral blood mononuclear cells of cancer patients with the HLA-A11 and HLA-A31 alleles more efficiently than the other 3 peptides. Antibody blocking and cold inhibition experiments revealed that the cytotoxicity of peptide-induced CTLs against cancer cells was attributable to peptide-specific and CD8+ T cells. Together with our previous findings, these results indicate that the 2 IEX peptides could be appropriate vaccine candidates for HLA-A11, HLA-A31, and HLA A33 positive epithelial cancer patients. This information could expand the chance of a peptide-based cancer vaccine for epithelial cancer patients of many ethnic populations. PMID- 17414319 TI - Identification and characterization of a WT1 (Wilms Tumor Gene) protein-derived HLA-DRB1*0405-restricted 16-mer helper peptide that promotes the induction and activation of WT1-specific cytotoxic T lymphocytes. AB - Effective tumor vaccine may be required to induce both cytotoxic T lymphocyte (CTL) and CD4+ helper T-cell responses against tumor-associated antigens. CD4+ helper T cells that recognize HLA class II-restricted epitopes play a central role in the initiation and maintenance of antitumor immune responses. The Wilms tumor gene WT1 is overexpressed in both leukemias and solid tumors, and the WT1 protein was demonstrated to be an attractive target antigen for cancer immunotherapy. In this study, we identified a WT1 protein-derived 16-mer peptide, WT1(332)(KRYFKLSHLQMHSRKH), which was restricted with HLA-DRB1*0405, one of the most common HLA class II types in Japanese, as a helper epitope that could elicit WT1-specific CD4+ T-cell responses. We established a WT1(332)-specific CD4+ helper T-cell clone (E04.1), which could respond to both HLA-DRB1*0405-positive, WT1-expressing transformed hematopoietic cells and autologous dendritic cells pulsed with apoptosis-induced WT1-expressing cells, indicating that the WT1(332) was a naturally processed helper epitope. Stimulation of peripheral blood mononuclear cells with both the CTL epitope (WT1(235)) and the helper epitope (WT1(332)) in the presence of WT1(332)-specific TH1-type CD4+ T cell clone strikingly enhanced the induction and the functional activity of WT1(235) specific CTLs compared with that of peripheral blood mononuclear cells with the WT1(235) alone. These results indicated that a helper epitope, WT1(332) should be useful for improvement of the efficacy of CTL epitope-based cancer vaccine targeting WT1 in the clinical setting. PMID- 17414320 TI - IL-2 and IL-15 each mediate de novo induction of FOXP3 expression in human tumor antigen-specific CD8 T cells. AB - Although FOXP3 is primarily expressed by regulatory CD4 T cells (Treg) in vivo, polyclonal activation of human CD8 T cells can result in the expression of FOXP3 in a fraction of CD8 T cells. However, the cellular lineage and mechanism of FOXP3 induction in CD8 T cells remain unclear. Here, we demonstrate that interleukin-2 (IL-2) induces FOXP3 expression in OKT3-stimulated or antigen stimulated CD8 T cells, indicating that FOXP3 expression is neither limited to a unique subset of CD8 T cells nor dependent on the mode of T-cell receptor stimulation. In the absence of IL-2, antigen stimulation resulted in T-cell activation and acquisition of effector function without induction of FOXP3, indicating that acquisition of effector function is independent of induction of FOXP3 expression in CD8 T cells. Interestingly, IL-15, but not IL-7 or IL-21, also led to de novo induction of FOXP3 in antigen-specific CD8 T cells, suggesting that signaling by IL-2/IL-15Rbeta chain is pivotal for induction of FOXP3 in human CD8 T cells. These findings indicate that induction of FOXP3 is intrinsic to CD8 T cells that are activated in the presence of IL-2 or IL-15, and in vitro-induced expression of FOXP3 cannot be simply interpreted as an indicator of Treg activity or activation marker. PMID- 17414321 TI - Monocytes immunoselected via the novel monocyte specific molecule, CD300e, differentiate into active migratory dendritic cells. AB - Monocytes, immunoselected using MMRI-1, a monoclonal antibody specific for CD300e, were used to generate dendritic cells (DC). These CD300e immunoselected monocyte-derived DC (MoDC) were compared phenotypically and functionally to CD14 immunoselected MoDC. CD300e and CD14 immunoselected mature MoDC expressed similar levels of the DC marker, CD83 and costimulatory molecules, CD80, CD86, and CD40. Both preparations took up soluble antigen with similar efficiency by pinocytosis and receptor mediated uptake. The CD300e and CD14 immunoselected MoDC also induced comparable CD4+ T lymphocyte allogeneic responses and recall responses to tetanus toxoid. Similar magnitude CD8 T lymphocyte responses to the naive antigen, MART-1 and the recall antigen, FMP, were induced by both MoDC preparations. Cytokine secretion by each type of MoDC preparation was similar; each secreted interleukin-12, tumor necrosis factor-alpha, and low levels of interferon-gamma but in most cases no interleukin-10. Migration studies confirmed that both types of MoDC migrated towards the chemokine, CCL21 although CD300e immunoselected showed greater migration. Overall, the CD14 immunoselected MoDC had higher spontaneous background migration, compared with the CD300e immunoselected MoDC. Differential signaling from the antibodies used to immunoselect the monocytes may account for the slight differences in migratory capacity. These data identify the CD300e antigen as another monocyte-specific marker that can be used to purify monocytes for differentiation into functionally active MoDC. PMID- 17414322 TI - Efficient chemokine-dependent migration and primary and secondary IL-12 secretion by human dendritic cells stimulated through Toll-like receptors. AB - Recent findings have demonstrated the properties of cell migration and cytokine secretion to be mutually exclusive and linked them to different functional subpopulations of dendritic cells (DCs). We studied human monocyte-derived DCs after stimulation with peptidoglycan (PGN), poly(I:C), lipopolysaccharide (LPS), and R848 (resiquimod) and found the resulting mature DCs to express CCR7, to migrate toward CCL19 and to be efficient primary interleukin (IL)-12 producers. Importantly, the potential for secondary production of large amounts of IL-12p70 in response to CD40 ligation was also preserved after stimulation by all Toll like receptor (TLR) ligands. Differences between the TLR ligands were seen in the primary secretion of IL-12 and IL-23, in the survival of the DCs and in the expression of CD38. Finally, DCs stimulated by R848 were efficient in expanding peptide-specific CD8-positive T cells capable of peptide-specific target cell lysis. Together, our data suggest that TLR ligands induce the generation of mature DCs that integrate migratory and cytokine secretory capacity as well as cytotoxic T lymphocyte (CTL) stimulatory properties. PMID- 17414323 TI - Resting B cells suppress tumor immunity via an MHC class-II dependent mechanism. AB - Resting B cells have been variously shown to induce direct tolerance of antigen specific CD8+ T cells, induce T-cell anergy via transforming growth factor-beta production, down-regulate interleukin-12 production by dendritic cells (DC) and influence TH1/TH2 differentiation via the production of regulatory cytokines. Through these mechanisms, B cells can exert a regulatory function in in vivo models of T-cell immunity including, experimental autoimmune encephalitis and rheumatoid arthritis. Here, we show that the resting B cells inhibit the ability of DC vaccination to provide protection from tumor growth. Inhibition of DC induced immunity by B cells was independent of presentation of major histocompatibility molecule (MHC) class-I bound tumor antigen but dependent on B cell expression of MHC class-II. Administration of B cells did not alter the ability of DC to migrate from the injection site or impair DC-T cell interactions within the draining lymph node. The inhibitory effect of B cells was lost when they were activated by CD40L and partially reversed by the depletion of CD4+/CD25+ regulatory T cells. Together our findings indicate that the resting B cells are capable of limiting CD8+ T-cell effector function induced by DC vaccination via a mechanism that is dependent on the expression of MHC class-II molecules. PMID- 17414324 TI - Pretreatment levels of sTNF-R1 and sIL-6R are associated with a higher vulnerability for IFN-alpha-induced depressive symptoms in patients with malignant melanoma. AB - Immunomodulatory therapy with interferon-alpha (IFN-alpha) often leads to neuropsychiatric side effects, especially depression. An activation of the immune system is discussed to trigger neurotransmitter changes and depressive illness. So far, few data are available about biologic markers, who may predict the individual risk for developing depressive symptoms during IFN-alpha therapy. The aim of the present study was to investigate the predictive role of certain immunologic markers for the development of IFN-alpha-induced depression. We hypothesized that patients characterized by a proinflammatory and TH1-accentuated immune response before treatment might have an increased risk for developing depressive mood changes. Thirty-three melanoma patients were prospectively investigated during adjuvant treatment with IFN-alpha-2a/2b (3 x 3 Mio units/wk). Depressive mood changes were assessed with the self-rating depression scale (SDS, Zung-scale) before and during IFN-alpha treatment. Serum concentrations of soluble tumor necrosis factor-R1 (sTNF-R1), soluble interleukin-6R (sIL-6R), sIL 4R, and neopterin were measured before and after 3 months of treatment. sIL-6R, which was negatively associated with SDS scores, significantly predicted higher depression scores in the first 3 months of IFN-alpha treatment. sTNF-R1, which was positively associated with SDS scores, significantly predicted the development of late depressive symptoms after 6 months of therapy. In contrast to the initial hypothesis, patients characterized by high sTNF-R1 and low sIL-6R baseline levels, indicating an anti-inflammatory condition before therapy, had a higher vulnerability for depression during IFN-alpha therapy. PMID- 17414325 TI - One step generation of fully chimeric antibodies using Cgamma1- and Ckappa mutant mice. AB - Humanized antibodies (Abs) are effective drugs against a variety of diseases such as cancer, autoimmune diseases, transplant rejection and others. The most powerful technology to develop humanized Abs is the use of mice that produce humanized Abs. By modifying the genetic background of F004 mice a new mouse substrain was developed for optimized "one step" generation of chimeric humanized monoclonal Abs. The new mice (F004-Jen) demonstrated improved fertility still expressing the human locus at the same level as the parental F004 mouse. The value of these mice for the generation of chimeric Abs was exemplified for a panel of chimeric Abs against the human neural cell adhesion molecule (NCAM): The fully chimeric human IgG1/kappa Ab Ch.MK1 bound to NCAM expressing cells with a K(D)=4.3-8.7 x 10(-8) M and was functionally active as demonstrated by depleting NCAM expressing cells. We also demonstrated that chimeric IgG1/kappa Abs can be induced by hybridoma class switching of IgM producing hybridoma cells, providing an alternative way to chimeric Abs. The present data highlight F004-Jen mice as an efficient tool for "one step" generation of chimeric Abs. PMID- 17414326 TI - Recognition of carcinoembryonic antigen peptide and heteroclitic peptide by peripheral blood T lymphocytes. AB - The carcinoembryonic antigen (CEA)-derived peptide CAP1 and heteroclitic peptide CAP1-6D are stimulators of HLA-A*A0201 restricted CEA-specific T cells in vivo and in vitro. The goal of this study was to evaluate differences in T cell responses to peptide and modified peptide antigens from CEA. The heterogeneity of responses among individuals is potentially important for the design of future CEA directed immunotherapy trials. Peripheral blood mononuclear cells from blood donors were stimulated with peptide, IL-2, and IL-7. Weekly, microcultures were restimulated with irradiated, autologous peptide-loaded peripheral blood mononuclear cells and expanded in IL-2. Established T cell lines were tested by cytokine release assays using peptide-loaded T2 targets. T cell avidity was measured by cytokine release using targets expressing diminishing concentrations of peptide. Fine specificities were measured using targets loaded with alanine substituted CAP1 peptide. Tumor recognition was measured using HLA-A*A0201/CAP1 transduced COS tumor targets. Varied responses to CAP1 and CAP1-6D were seen among individuals. The immunogenicity of CAP1 or CAP1-6D was donor dependent. Many T cells recognized one peptide but did not cross-recognize the altered peptide. The avidities of T cell lines were moderate to low, and fine specificities were consistent with a narrow antigen-specific repertoire. CAP1-6D based immune therapy may not be optimal in some patients with CAP1-specific precursors. The T cell repertoire may be a central contributor to the limited responses seen with CEA-directed immunotherapy to date. Treatment strategies designed to alter or expand the T cell repertoire against CEA should be considered for trials. PMID- 17414327 TI - The acute care surgery curriculum. PMID- 17414328 TI - The practice of venous thromboembolism prophylaxis in the major trauma patient. AB - BACKGROUND: The incidence of venous thromboembolism (VTE) without prophylaxis is as high as 80% after major trauma. Initiation of prophylaxis is often delayed because of concerns of injury-associated bleeding. As the effect of delays in the initiation of prophylaxis on VTE rates is unknown, we set out to evaluate the relationship between late initiation of prophylaxis and VTE. METHODS: Data were derived from a multicenter prospective cohort study evaluating clinical outcomes in adults with hemorrhagic shock after injury. Analyses were limited to patients with an Intensive Care Unit length of stay >or=7 days. The rate of VTE was estimated as a function of the time to initiation of pharmacologic prophylaxis. A multivariate stepwise logistic regression model was used to evaluate factors associated with late initiation. RESULTS: There were 315 subjects who met inclusion criteria; 34 patients (11%) experienced a VTE within the first 28 days. Prophylaxis was initiated within 48 hours of injury in 25% of patients, and another one-quarter had no prophylaxis for at least 7 days after injury. Early prophylaxis was associated with a 5% risk of VTE, whereas delay beyond 4 days was associated with three times that risk (risk ratio, 3.0, 95% CI [1.4-6.5]). Factors associated with late (>4 days) initiation of prophylaxis included severe head injury, absence of comorbidities, and massive transfusion, whereas the presence of a severe lower extremity fracture was associated with early prophylaxis. CONCLUSIONS: Clinicians are reticent to begin timely VTE prophylaxis in critically injured patients. Patients are without VTE prophylaxis for half of all days within the first week of admission and this delay in the initiation of prophylaxis is associated with a threefold greater risk of VTE. The relative risks and benefits of early VTE prophylaxis need to be defined to better direct practice in this high-risk population. PMID- 17414329 TI - Thromboembolic complications associated with factor VIIa administration. AB - BACKGROUND: Factor VIIa (FVIIa) is given to promote hemostasis in coagulopathic trauma patients at high risk for death. FVIIa is associated with thromboembolic complications, but the incidence is not known, nor is which patients are at risk. We examined our experience with FVIIa to better understand this issue. METHODS: Medical records of patients who received FVIIa from 2001 to 2006 were reviewed for evidence of thromboembolic events, including ischemic cardiovascular accident, myocardial ischemia or infarct, mesenteric infarct, peripheral arterial thromboembolism, deep venous thrombosis, or pulmonary embolus. Indication for FVIIa dosing, timing of complication, and clinical outcome were recorded. Each event was assessed by a panel of experienced clinicians to determine the contribution of FVIIa to the event and to patient outcome. RESULTS: Two hundred eighty-five medical records were reviewed. Twenty-seven patients (9.4%) had thromboembolic complications after administration of FVIIa. Nine events were thought to be highly related to FVIIa, and 10 of 14 deaths were, in part, caused by the thrombotic complication. Eighteen of the events, including all cardiovascular accidents and most cases of mesenteric ischemia, were attributed to a combination of FVIIa and a definable, high-energy vascular injury. CONCLUSION: FVIIa is a potent procoagulant, with the potential for adverse thromboembolic events in susceptible patients. Our experience suggests that caution should be exercised in administration of FVIIa to patients with arterial injuries. Injured mesenteric and cerebral vessels may be especially susceptible to thrombosis. PMID- 17414330 TI - Extracorporeal life support for massive pulmonary embolism. AB - BACKGROUND: Massive pulmonary embolism is frequently lethal because of acute irreversible pulmonary and cardiac failure. Extracorporeal life support (ECLS) has been used for cardiopulmonary failure in our institution since 1988, and we reviewed our experience with its use in the management of massive pulmonary emboli. METHODS: We reviewed our complete experience with ECLS for massive pulmonary emboli from January 1992 through December 2005. The records of 21 patients were examined and data extracted. RESULTS: During the study period, 21 patients received ECLS for massive pulmonary emboli. All patients were on vasoactive drugs, acidemic, and hypoxic at the time of institution of ECLS. Eight were in active cardiac arrest. Five were trauma patients, eight had recently undergone an operation, and six had a hypercoagulable disorder. Nineteen of the 21 patients were cannulated for venoarterial bypass and two were placed on venovenous bypass. The average duration of support for survivors was 5.4 days, ranging from 5 hours to 12.5 days. Emboli resolved with anticoagulation in 10 of 13 survivors and 4 of 13 survivors underwent surgical pulmonary embolectomy. Catastrophic neurologic events were the most common cause of mortality in our series; four patients died from intracranial hemorrhage. The overall survival rate was 62% (13/21). CONCLUSIONS: We conclude that emergent ECLS provides an opportunity to improve the prognosis of an otherwise near-fatal condition, and should be considered in the algorithm for management of a massive pulmonary embolism in an unstable patient. PMID- 17414331 TI - Trauma in adolescents causes long-term marked deficits in quality of life: adolescent children do not recover preinjury quality of life or function up to two years postinjury compared to national norms. AB - BACKGROUND: Injury is a leading cause of death and preventable morbidity in adolescents. Little is known about long-term quality of life (QoL) outcomes in injured adolescents. The objectives of the present report are to describe long term QoL outcomes and compare posttrauma QoL to national norms for QoL in uninjured adolescents from the National Health Interview Survey (NHIS). METHODS: In all, 401 trauma patients aged 12 to 19 years were enrolled in the study. Enrollment criteria excluded spinal cord injury. QoL after trauma was measured using the Quality of Well-being (QWB) scale, a sensitive and well-validated functional index (range: 0 = death to 1.000 = optimum functioning). Patient outcomes were assessed at discharge, and 3, 6, 12, 18, and 24 months after discharge. NHIS data were based on 3 survey years and represent a population based U.S. national random sample of uninjured adolescents. RESULTS: Major trauma in adolescents was associated with significant and marked deficits in QoL throughout the 24-month follow-up period, compared with NHIS norms for this age group. Compared with NHIS norms for QoL in uninjured adolescents aged 12 to 19 years (N = 81,216,835; QWB mean = 0.876), injured adolescents after major trauma had striking and significant QoL deficits beginning at 3-month follow-up (QWB mean = 0.694, p < 0.0001), that continued throughout the long-term follow-up 24 months after discharge (6-month follow-up QWB mean = 0.726, p < 0.0001; 12-month follow-up QWB mean = 0.747, p < 0.0001; 18-month follow-up QWB mean = 0.758, p < 0.0001; 24-month follow-up QWB mean = 0.766, p < 0.0001). QoL deficits were also strongly associated with age (>or=15 years) and female sex. Other significant risk factors for poor QoL outcomes were perceived threat to life, pedestrian struck mechanism, and Injury Severity Scores >16. CONCLUSIONS: Major trauma in adolescents is associated with significant and marked deficits in long-term QoL outcomes, compared with U.S. norms for healthy adolescents. Early identification and treatment of risk factors for poor long-term QoL outcomes must become an integral component of trauma care in mature trauma care systems. PMID- 17414332 TI - Emergency room management of patients with blunt major trauma: evaluation of the multislice computed tomography protocol exemplified by an urban trauma center. AB - BACKGROUND: The early clinical management of patients with major trauma still represents a challenge. To clinically evaluate the full extent of a patient's injuries is difficult, especially when the patient is unconscious. Before December 2002, trauma patients admitted to our emergency room (ER) underwent a diagnostic protocol including physical examination, conventional radiography (CR), sonography and further procedures if necessary. After the installation of a MSCT scanner, all trauma patients underwent the "MSCT protocol" immediately after admission. The aim of the study was to compare the "MSCT-protocol" as it is performed at our institution, with the "Pre-MSCT-protocol". METHODS: We compared 185 patients undergoing the "Pre MSCT-protocol" with 185 patients undergoing "MSCT protocol". We evaluated the efficacy, speed and accuracy of the "MSCT protocol" using several variables. Time periods from admission to the ER to admission to the intensive care unit were compared as well as outcome parameters such as length of ICU stay, ventilation period and rates of organ. Dichotomous data were analyzed by Chi-square analysis; continuous data were analyzed by Student's t test. Any values of p < 0.05 were considered significant for any test. RESULTS: No significant differences were found regarding demographic data. The full extent of injuries was definitively diagnosed after 12 +/- 9 minutes in 92.4% of the "MSCT protocol" cohort. In only 76.2% of "Pre-MSCT protocol" cohort definitive diagnosis was possible after 41 +/- 27 minutes. Total ER time was 104 +/- 21 minutes with the "Pre-MSCT protocol" and 70 +/- 17 minutes with "MSCT protocol" (p < 0.05). "Pre-MSCT protocol" patients had a significantly longer ICU stay than "MSCT protocol" patients (p < 0.05). "MSCT protocol" patients had significantly fewer ventilation days (14.3 vs. 10.9 days). Furthermore, rates of organ failure were lower in patients undergoing the "MSCT protocol". CONCLUSION: We could demonstrate that immediate MSCT in patients with blunt major trauma leads to more accurate and faster diagnosis, and reduction of early clinical time intervals. We also observed a reduction in ventilation, ICU, and hospital days, and in organ failure rates, though this might have been partly due to small differences in case mix. The "MSCT protocol" algorithm seems to be safe and effective. PMID- 17414333 TI - Predicting hospital mortality, length of stay, and transfer to long-term care for injured patients. AB - BACKGROUND: Using hospital length of stay (LOS) to measure trauma care efficiency is complicated by short LOS resulting from early mortality or transfer to long term care (LTC). METHODS: Records from the 1999 to 2003 National Trauma Data Bank were used to create a multistate model divided into four time periods, each with constant rates of death, discharge home, and LTC transfer. Estimated hospital mortality and LOS for patient subgroups were calculated from this model, and time varying covariate effects were estimated. RESULTS: A total of 369,829 cases with adequate data were available. Early mortality was increased most by hypotension or coma, and also by anatomic injury severity or penetrating mechanism, but these effects diminished with time; age remained a strong predictor of mortality at any time but sex was insignificant. Rates of discharge home decreased with time, whereas rates of LTC transfer peaked at 6 to 11 days. Increased age strongly predicted transfer to LTC, whereas penetrating or burn mechanisms made it less likely. Predicted and observed outcomes were similar for multiple subgroups, and about 17% of individual variation in LOS was explained by the model. CONCLUSIONS: Multistate models of patient status can accurately predict mortality and resource use after injury, and describe time-varying effects of other factors. PMID- 17414334 TI - Modeling probability-based injury severity scores in logistic regression models: the logit transformation should be used. AB - BACKGROUND: The International Classification of Disease Injury Severity Score (ICISS) and the Trauma Registry Abbreviated Injury Scale Score (TRAIS) are trauma injury severity scores based on probabilities of survival. They are widely used in logistic regression models as raw probability scores to predict the logit of mortality. The aim of this study was to evaluate whether these severity indicators would offer a more accurate prediction of mortality if they were used with a logit transformation. METHODS: Analyses were based on 25,111 patients from the trauma registries of the four Level I trauma centers in the province of Quebec, Canada, abstracted between 1998 and 2005. The ICISS and TRAIS were calculated using survival proportions from the National Trauma Data Bank. The performance of the ICISS and TRAIS in their widely used form, proportions varying from 0 to 1, was compared with a logit transformation of the scores in logistic regression models predicting in-hospital mortality. Calibration was assessed with the Hosmer-Lemeshow statistic. RESULTS: Neither the ICISS nor the TRAIS had a linear relation with the logit of mortality. A logit transformation of these scores led to a near-linear association and consequently improved model calibration. The Hosmer-Lemeshow statistic was 68 (35-192) and 69 (41-120) with the logit transformation compared with 272 (227-339) and 204 (166-266) with no transformation, for the ICISS and TRAIS, respectively. CONCLUSIONS: In logistic regression models predicting mortality, the ICISS and TRAIS should be used with a logit transformation. This study has direct implications for improving the validity of analyses requiring control for injury severity case mix. PMID- 17414335 TI - External validation and modification of a pediatric trauma triage tool. AB - BACKGROUND: Simon et al. developed a simple secondary triage tool (mPTS) based on physiologic parameters and physical findings to identify pediatric trauma patients who had a low likelihood of serious injury. Such patients could be treated in the emergency room without full trauma team activation. Our objective was to evaluate the mPTS on the trauma population at our institution, a Level I pediatric trauma center. METHODS: This was a retrospective cohort study of all trauma team activations at The Hospital for Sick Children (Sick Kids) (1999 2002), excluding penetrating trauma and burns. Patients were stratified into high risk (Injury Severity Score [ISS] >or=12) and low-risk (ISS <12) groups. The mPTS evaluates airway integrity, open wounds, neurologic status, hemodynamics, and skeletal integrity and applies a score of 1 point to each criterion. RESULTS: There were 628 trauma patients (382 boys, mean age of 8 +/- 3.8 years). The mPTS had a sensitivity of 92% and a positive predictive value (PPV) of 47% when applied to our population. The mPTS missed 21 patients with significant injuries, many were intra-abdominal. We modified the mPTS to include contusions to head and/or torso and a history of loss of consciousness and a 7-point score was developed. After modification the sensitivity was 99%, specificity 21%, and PPV of 46% with a 20% reduction in unnecessary trauma team activations. CONCLUSIONS: The original mPTS by Simon et al. was not sensitive enough when applied to our population. The Sick Kids modification to the score improved the sensitivity to 99%. The PPV of 46% indicates a safe level of overtriage is maintained. The Sick Kids mPTS remains easy to apply and would have reduced trauma team activation by 20%. PMID- 17414336 TI - Optimizing physician staffing and resource allocation: sine-wave variation in hourly trauma admission volume. AB - BACKGROUND: Trauma centers are faced with the challenge of managing an increasing volume of patients in an era of fewer trauma care providers and fewer hospitals providing trauma care. The purpose of this study was to determine the relationship between hourly admission volume, injury severity, resource utilization, and outcomes. METHODS: All patients in the National Trauma Data Bank admitted between 1999 and 2002 were selected. Analysis included demographics, temporal information, injury severity, and outcome parameters. RESULTS: A total of 421,997 patients were admitted to participating centers. The 24-hour admission distribution described a sine-wave pattern with a trough at 6:00 am and a peak at 7:00 pm. The sine-wave pattern persisted regardless of the subgroup analyzed. Patients admitted between 12:00 am and 6:00 am were more likely to be severely injured, require intensive care unit admission, undergo emergent operation, and die during hospitalization when compared with patients admitted between 7:00 am and 12:00 pm. CONCLUSIONS: Trauma admissions conform to a sine-wave pattern with a 3.5-fold increase in admissions between morning and evening hours. This has significant implications for manpower and resource allocation with additional resources needed in the hours around 7:00 pm and later. Educational and administrative activities are best scheduled during low-volume morning hours. Nighttime admissions are higher risk and thus more likely to need senior-level expertise and consume hospital resources. Trauma centers should use these findings to improve outcomes by developing optimal staffing patterns and matching resource allocation to need as a function of time. PMID- 17414337 TI - Trauma center downstream revenue: the impact of incremental patients within a health system. AB - BACKGROUND: The purpose of this study is to assess the downstream clinical and financial impact of a trauma, burn, and emergency surgery service at an academic Level I trauma center. METHODS: All patients admitted to the trauma, burn, and emergency surgery service from fiscal years 2002 to 2004 were identified. Clinical and financial data including inpatient and outpatient activity were analyzed for 365 days (downstream) after initial service admission. Data were divided into total service, trauma and burn, inpatient, outpatient, hospital, and professional revenue. RESULTS: In all, 3,679 patients were admitted during the study period with total initial revenue approaching $103 million. Of these, 1,566 patients were subsequently admitted for downstream inpatient activity, resulting in almost $26 million in subsequent inpatient revenue. The initial patient admissions resulted in over 17,000 clinic visits during the course of the 3 study years. Professional revenue resulted in over $14 million for the initial admission and $6.1 million in downstream revenue during the study period. CONCLUSIONS: Trauma, burn, and emergency surgical services result in both substantial initial and downstream revenue for the hospital (inpatient and outpatient) and professional components. Services committed to caring for the injured and emergent patients substantially contribute to the institutional financial strength. PMID- 17414338 TI - The direct economic burden of blunt and penetrating trauma in a managed care population. AB - BACKGROUND: Although the prevalence of trauma in the United States is high, data on the economic burden of this public health problem to third-party payors is limited. METHODS: Retrospective claims from a large health plan were analyzed for 12,615 adults (age >or=18 years) hospitalized for blunt or penetrating trauma between January 1, 2003 and February 1, 2005. Per patient charges were estimated for resources utilized during a 6-month period before and after initial injury. Continuous health plan enrollment during these periods was required. Three cohorts were examined: isolated traumatic brain injury (TBI); other trauma with TBI (trauma w/TBI); and other trauma without TBI (trauma w/o TBI). Patients were also stratified by Injury Severity Score (ISS) and trauma designation of the admitting hospital. RESULTS: Initial hospitalization charges ranged from $32,627 for isolated TBI to $103,667 for trauma w/TBI. Charges for initial hospitalization were highest ($199,443) among patients with the most severe injuries. Overall, initial hospitalization charges were highest among those admitted to Level I trauma centers ($68,626); for trauma w/TBI, however, initial hospitalization charges were highest among those admitted to nontrauma centers ($130,997). Charges incurred during postdischarge medical encounters ranged from $16,361 for isolated TBI to $23,761 for trauma w/TBI. Increased charges for postdischarge encounters compared with the 6-month preinjury period ranged from $6,756 for isolated TBI to $19,771 for trauma w/TBI. CONCLUSIONS: The economic burden of blunt and penetrating trauma to third-party payors is high. Efforts to reduce the incidence of trauma may result in substantial economic savings to managed care systems. PMID- 17414339 TI - The increasing use of chest computed tomography for trauma: is it being overutilized? AB - OBJECTIVE: Chest computerized tomography (CCT) has gained popularity in trauma evaluation, but it is expensive, increases exposure to radiation, and leads to findings of debatable clinical significance. The objective of this study was to determine the change in usage of CCT over time and the number of injuries missed on plain chest radiograph (CXR) with normal findings that required therapy. METHODS: Data on all patients evaluated with a screening CXR during a 7-year period were extracted from a Level I center trauma registry. The incidence of CCT utilization during the duration of the study was identified. Patients who had CCT done after the initial CXR were analyzed separately for the presence of occult injuries. The association of these findings with demographic and injury data were examined. RESULTS: There were 2,326 CCT performed, and 1,873 (80.5%) of them were after negative CXRs. The percentage of patients studied with CCT increased incrementally from 2.7% to 28.7% for blunt and from 0.4% to 2.9% for penetrating injury. The identification of occult pneumothorax, hemothorax, rib fractures, and lung contusions significantly increased during the study period with the increased frequency of CCT use. There were 102 occult pneumothoraces and/or hemothoraces identified, but only 12 patients underwent tube thoracostomy during the 7-year period. There were 43 patients with blunt aortic injury (BAI) and 6 (13.9%) of these patients had normal CXR findings. There was no trend in increased BAI diagnosed during the study period, although the utilization of CCT was increased. CONCLUSIONS: There has been a 10-fold increase in use of CCT for trauma evaluation. Although occult findings increased, the number of patients who needed treatment was small. The excess utilization of CCT after negative CXR needs continued refinement to identify the small number of potentially lethal injuries while reducing the number of trivial findings. PMID- 17414340 TI - Lactated Ringer's is superior to normal saline in the resuscitation of uncontrolled hemorrhagic shock. AB - BACKGROUND: Normal saline (NS) and lactated Ringer's solution (LR) continue to be used interchangeably for the resuscitation of hemorrhagic shock in some institutions. We hypothesized that, aside from hyperchloremic acidosis, NS resuscitation would be similar to that of LR in a swine model of uncontrolled hemorrhage. METHODS: Twenty swine weighing a mean of 37 kg underwent invasive line placement, midline celiotomy, and splenectomy. After a 15-minute stabilization period, we recorded a baseline mean arterial pressure (MAP) and created a grade V liver injury. The animals bled freely for 30 minutes after which we measured blood loss. We blindly randomized the swine to receive NS (10 animals) versus LR (10 animals) to achieve and maintain the baseline MAP for 90 minutes postinjury. Laboratory values were obtained at baseline and upon completion of the 2-hour study period. RESULTS: Initial blood loss was 25 mL/kg in the NS group and 22 mL/kg in the LR group (p = 0.54). Animals required 256.3 +/- 145.4 mL/kg of fluid in the NS group as compared with 125.7 +/- 67.3 mL/kg in the LR group (p = 0.04). The urine output was higher in the NS group (46.6 +/- 39.5 mL/kg versus 18.9 +/- 12.9 mL/kg, p = 0.04). Upon study completion, the NS group had a significant hyperchloremia (119 +/- 1.9 mEq/L versus 105 +/- 2.9 mEq/L, p < 0.01) with acidosis (7.28 +/- 0.12 versus 7.45 +/- 0.06, p < 0.01) in comparison to the LR group. In addition, resuscitation with NS resulted in significantly lower fibrinogen levels (99 +/- 21 mg/dL versus 123 +/- 20 mg/dL, p = 0.02). The serum lactate was 4.7 +/- 2.2 in the LR group and 1.7 +/- 1.7 in the NS swine (p < 0.01) at the end of the study. CONCLUSIONS: Resuscitation of uncontrolled hemorrhagic shock with NS requires significantly greater volume and is associated with greater urine output, hyperchloremic acidosis, and dilutional coagulopathy as compared with LR. Resuscitation with LR results in an elevation of the lactate level that is not associated with acidosis. Lactated Ringer's solution is superior to NS for the resuscitation of uncontrolled hemorrhagic shock in swine. PMID- 17414341 TI - Small volume resuscitation: a randomized controlled trial with either norepinephrine or vasopressin during severe hemorrhage. AB - BACKGROUND: The present study was designed to evaluate the effects of hypertonic hyperoncotic hydroxyethyl starch solution (HHS) combined with either norepinephrine (NE) or arginine vasopressin (AVP) on cerebral perfusion pressure (CPP) and brain metabolism after hemorrhagic shock. METHODS: Fourteen pigs were subjected to uncontrolled liver bleeding until hemodynamic decompensation followed by resuscitation using HHS (4 mL/kg) combined with either NE (bolus of 1000 microg; 60 microg/kg/hr; n = 7) or AVP (bolus of 10 U; 2 U/kg/hr; n = 7), respectively. Extracellular cerebral concentrations of glucose, glycerol, lactate, and the lactate/pyruvate ratio were assessed by microdialysis. After 30 minutes of therapy, bleeding was controlled by manual compression and all surviving animals were observed for 1 hour. RESULTS: After hemodynamic decompensation, AVP resulted in a significantly higher increase of CPP (mean +/- SD; 47 +/- 19 versus 28 +/- 9 mm Hg; p < 0.01) and cerebral venous partial pressure of oxygen (66 +/- 8 versus 49 +/- 9 mm Hg; p < 0.05) compared with NE after 10 minutes of therapy. Hemodynamic data and blood gas variables were not different between groups during the remaining study period. Brain metabolism was found comparable in both groups at any time. CONCLUSIONS: AVP was comparable to NE with respect to hemodynamics and blood gases, as well as brain metabolism in surviving animals throughout the study period. Our findings emphasize the importance of early resuscitation, as neuronal cell damage potentially starts immediately after onset of severe hemorrhage. PMID- 17414342 TI - Increased intra-abdominal, intrathoracic, and intracranial pressure after severe brain injury: multiple compartment syndrome. AB - OBJECTIVES: Fluid therapy and/or acute lung injury may increase intra-abdominal pressure (IAP) and intrathoracic pressure, thereby increasing intracranial pressure (ICP) after traumatic brain injury (TBI). Further fluid administration to support cerebral perfusion or increasing ventilatory support to treat acute lung injury further increases ICP. This can create a cycle that ultimately produces multiple compartment syndrome (MCS). Both decompressive craniectomy (DC) and decompressive laparotomy (DL) decrease ICP. DL can also decrease IAP and ICP. We evaluated the serial application of DC and DL to treat MCS. METHODS: Data were analyzed for 102 consecutive patients with severe TBI who underwent DC alone to decrease ICP or in combination with DL to treat MCS. RESULTS: All 102 patients sustained blunt injury. Seventy percent were men with a mean age of 29.5 years, an Injury Severity Score of 34.4, and admission Glasgow Coma Scale score of 7.1. Fifty-one patients had diffuse brain injury and 51 had mass lesions. Seventy eight patients (76%) underwent DC alone. Twenty-four (22%) had both therapies for MCS. Fifteen patients had DC before DL and nine had DL before DC. Mean time between DC and DL was 3.4 +/- 6 days. The mean IAP before DL was 28 +/- 5 mm Hg. Twenty-four-hour cumulative mean intrathoracic pressure decreased significantly after DL in the MCS group (p = 0.01). Mean ICP decreased significantly after both DC and DL (p < 0.05). CONCLUSION: Increased ICP may be from primary TBI or MCS. Patients with MCS have a higher Injury Severity Score, ICP, and fluid requirements, but no increase in mortality. Both DC and DL reduce ICP and can be used in sequence. MCS should be considered in multiply injured patients with increased ICP that does not respond to therapy. PMID- 17414343 TI - The association between the postconcussion symptoms and clinical outcomes for patients with mild traumatic brain injury. AB - BACKGROUND: Postconcussion symptoms (PCS) (such as dizziness, headache, irritability, fatigue, and impaired memory) are common in patients who sustain a mild traumatic brain injury (mTBI). However, few studies have systematically investigated the association between PCS and clinical outcomes in mTBI patients. Therefore, the present study attempted to examine PCS during the disease course and to determine whether PCS adversely affect outcome. METHODS: This was a prospective, cohort and controlled study of 115 mTBI patients. The PCS checklist was used to identify PCS and the Glasgow Outcome Scale and the Glasgow Outcome Scale-Extended were used to investigate clinical outcomes. All patients were evaluated four times: at 1 week, 2 weeks, 4 weeks, and 8 weeks after the injury. RESULTS: Physical symptoms such as dizziness and headache were prominent in the early after injury stage (1 and 2 weeks). On the other hand, the psychosocial symptoms, such as depression and irritability, were significant at the late after injury stage (4 and 8 weeks). Dizziness adversely affected clinical outcome at both the early and late stages of the disease, whereas the impact of intracranial lesions and depression on outcome was greatest early and late, respectively. CONCLUSIONS: The results show that PCS during the disease course and the relationship between PCS and clinical outcome can be systematically evaluated. In fact, different postconcussion symptom domains should be monitored while the disease is progressing. PMID- 17414344 TI - Incidence and crash mechanisms of aortic injury during the past decade. AB - BACKGROUND: Aortic injuries were traditionally thought to be the result of severe frontal crashes. Newer data has suggested other crash types such as nearside crashes may also be important in aortic injury. We hypothesized the implementation of recent safety measures would decrease the incidence of aortic injury associated with fatal motor vehicle crashes. METHODS: The autopsy reports of all traffic fatalities for motor vehicle occupants in a large urban county for the years 1993 to 2004 were examined. The demographics, impact types, safety measures used, and the presence of any aortic injury were recorded. Trends were evaluated for significance by weighted linear regression. RESULTS: The incidence of aortic injury associated with fatal motor vehicle crashes has remained unchanged during the past 12 years (r = 0.057, p = 0.45). There is a trend toward decreased aortic injuries associated with frontal crashes (r = 0.26, p = 0.089) but no change in aortic injuries associated with nearside or farside crashes (r = 0.053, p = 0.47), when the crash resulted in a fatality. This is despite an increase in seat belt use and increased presence of airbags during the same time period. CONCLUSIONS: Despite improved safety measures designed to minimize the occurrence of aortic injuries, the incidence of blunt aortic injury in fatal motor vehicle crashes has not decreased during the past decade. Although not statistically significant, there is a trend toward decreased frontal impacts in fatal motor vehicle crashes associated with aortic injuries. The nearside crash mechanism continues to play a prominent role, and efforts at improving vehicle safety should be focused on crash mechanisms as they relate to aortic injury. PMID- 17414345 TI - Superior mesenteric venous injuries: to ligate or to repair remains the question. AB - BACKGROUND: Superior mesenteric vein injuries are rare and incur high mortality. Given their low incidence, little data exist delineating indications for when to institute primary repair versus ligation. The purposes of this study are to review our institutional experience, to determine the additive effect on mortality of associated vascular injuries, to correlate mortality with the American Association for the Surgery of Trauma-Organ Injury Scale (AAST-OIS) for abdominal vascular injury and to examine and define the indications and outcomes for primary repair versus ligation. MATERIAL: Retrospective 156 months study (January 1992 through December 2004) in a large Level I urban trauma center of all patients admitted with superior mesenteric vein injuries. Patients were stratified, according to surgical technique employed to deal with their injuries, into those undergoing primary repair versus ligation to determine outcomes and define the surgical indications of these methods. The main outcome measure was overall survival. Cases of survival were stratified according to surgical method: primary repair versus ligation. RESULTS: There were 51 patients with a mean Injury Severity Score of 25 +/- 12. Mechanism of injury was penetrating for 38 (76%), blunt for 13 (24%), and patients undergoing emergency department thoracotomy for 4 (8%). Surgical management was ligation for 30 (59%), primary repair for 16 (31%), and 5 (10%) patients were exsanguinated before repair. The overall survival rate was 24/50 (47%). The survival rate excluding patients undergoing emergency department thoracotomy was 51%. The survival rate excluding patients that sustained greater than 3 to 4 associated vessels injured was 65%. The survival rates of patients with superior mesenteric vein and superior mesenteric artery was 55% and superior mesenteric vein and portal vein (PV) was 40%. The survival rate of patients with isolated superior mesenteric vein injuries was 55%. Mortality stratified to AAST-OIS grade III, 44%; grade IV, 42%; and grade V, 42%. Survival rates stratified to method of management consisted of primary repair (60%) versus ligation (40%). CONCLUSIONS: SMV injuries are highly lethal. Multiple associated vessel injuries increase mortality. Mortality correlates well with the American Association for the Surgery of Trauma-Organ Injury Scale for abdominal vascular injuries. Patients undergoing primary repair have higher survival rates (63%) and lesser numbers of associated vascular and nonvascular injuries; whereas those undergoing ligation have a smaller survival rate (40%) and higher number of associated vascular and nonvascular injuries. Ligation appears to be safe and should be selected for hemodynamically unstable patients with a large number of associated injuries. PMID- 17414346 TI - Propranolol does not increase inflammation, sepsis, or infectious episodes in severely burned children. AB - BACKGROUND: Propranolol, a nonselective beta1-2 antagonist, attenuates hypermetabolism and catabolism in severely burned patients. However, recent data suggest that propranolol impairs immune function and enhances inflammation. The purpose of the present study was to determine the effect of propranolol administration on infection, sepsis, and inflammation in severely burned pediatric patients. PATIENTS: A prospective, intent-to-treat study was performed; patient demographics (age, gender, burn size, and mortality); infectious episodes (colony count greater then 10); and sepsis (guidelines by the society of critical care medicine) were determined. Hypermetabolic response was determined by resting energy expenditure (REE), and the inflammatory response was determined by measuring serum cytokine expression. RESULTS: Two hundred forty-five patients (143 controls, 102 propranolol) were included into the study. There were no differences between the control and propranolol groups for age, gender distribution, burn size, third degree burn, and length of stay. Mortality was 6% in the control group and 5% in the propranolol group. Propranolol significantly decreased REE and predicted REE during acute hospital stay. Forty-three patients developed infections in the control group (30%), whereas 21 developed infections in the propranolol group (21%). The incidence of sepsis was 10% for controls and 7% for propranolol. Analysis of the cytokine expression profile in 20 patients in each group revealed that propranolol significantly decreased serum tumor necrosis factor and interleukin-1beta compared with controls (p < 0.05). CONCLUSION: Propranolol treatment attenuates hypermetabolism and does not cause increased incidence of infection and sepsis. PMID- 17414347 TI - Incidence and pattern of burn injuries among children with disabilities. AB - BACKGROUND: Burn injuries are an important cause of severe morbidity and mortality among children. However, the epidemiology of burns among disabled children has received little attention. METHODS: Burn injuries were identified for children aged less than 12 years using International Classification of Diseases, Ninth Revision, Clinical Modification codes in Ohio Medicaid claims data. Using FY2002 Ohio Medicaid claims data, incidences and relative risks of burn injuries for disabled and nondisabled children were calculated by age, gender, and race or ethnicity. Logistic regression was used to analyze risk factors for burn injuries. RESULTS: There were 4,307 burn injuries identified in the FY2002 Ohio Medicaid claims database. The incidence of burn injuries for disabled children was significantly higher than for nondisabled children (103.00 per 10,000 vs. 77.41 per 10,000, respectively; p < 0.001). Children aged 1 or 2 years had the highest incidence of burn injuries, regardless of disability status. For disabled children, the incidence of burn injuries decreased after 2 years of age and leveled out at approximately 100 per 10,000 children after 3 years of age. However, for nondisabled children, the incidence of burn injuries decreased until 6 years of age, after which it leveled out at approximately 40 per 10,000 children. After controlling for potentially confounding factors, the risk of burn injuries was significantly higher for disabled than nondisabled children (odds ratio = 1.80; 95% confidence interval, 1.50-2.17). CONCLUSIONS: Disabled children had a significantly higher incidence of burn injuries than nondisabled children did. The risk of burn injuries, even after controlling for demographic factors, was significantly higher for disabled children than nondisabled children. PMID- 17414348 TI - Associated injuries and not fracture instability predict mortality in pelvic fractures: a prospective study of 100 patients. AB - BACKGROUND: Whether pelvic fracture instability is correlated to mortality in blunt multiply-injured trauma patients is debatable. This is the first prospective study on patients with pelvic fractures aiming at finding whether pelvic fracture type affects mortality. METHODS: There were 100 consecutive patients (77 males, mean age of 31 [3-73] years) studied between September 2003 and October 2004. Data were collected regarding mechanism of injury, associated injuries, Injury Severity Score (ISS), Revised Trauma Score, blood transfusions, and mortality. The fractures were classified according to instability, where type O is stable, type R is rotationally unstable, and type RV is both rotationally and vertically unstable. Because a pure acetabular fracture is a single break in the pelvic ring, we classified it as type O. Computer tomography was used for fracture classification in 73 patients and plain X-rays in 27 patients. RESULTS: There were 77 fractures caused by road traffic collisions. Type O fractures (n = 63) had lower median ISS (13 [4-48]) than type R (n = 19) (18 [9-75]) and type RV (n = 18) (18 [6-66]) (p = 0.019, Kruskall Wallis). There was no significant difference in ISS between type R and RV fractures. A logistic regression model has shown that ISS was the only significant factor that predicts mortality. CONCLUSIONS: ISS is the most important predictor in defining mortality in patients with pelvic fracture and not the type of pelvic instability. PMID- 17414349 TI - Early unreamed intramedullary nailing of femoral fractures is safe in patients with severe thoracic trauma. AB - BACKGROUND: The timing of fixation of femoral fractures in multiply injured patients with severe thoracic trauma is discussed controversially. Some authors recommend damage control surgery, whereas other authors prefer early definitive treatment. The aim of our study was to investigate the effect of early definitive fixation of femoral fractures on outcomes in multiply injured patients with severe thoracic trauma. METHODS: Between May 1, 1998 and December 31, 2004, 578 severely injured patients were admitted to our institution. Forty-five patients met the inclusion criteria for the study cohort (severe thoracic trauma and femoral fracture stabilized with unreamed intramedullary nailing [IMN] within the first 24 hours) and 107 patients were selected for the control cohort (severe thoracic trauma without any lower extremity fracture). Inclusion criteria for both cohorts were age 15 to 55 years with blunt trauma (e.g. motor vehicle collisions, falls) including severe thoracic trauma (Abbreviated Injury Scale [AIS] score >or=3) and Injury Severity Score (ISS) >or=18. For comparison between the cohorts data on patients status (Glasgow Coma Scale score at arrival, Revised Trauma Score, Trauma and Injury Severity Score survival prognosis, Simplified Acute Physiology Score II score), treatment (intubation rate, thoracic drainage, surgery), and outcomes (duration of intensive care unit stay and ventilation, rate of adult respiratory distress syndrome [ARDS], multiple organ failure syndrome [MOFS], and mortality) were selected from hospital databases. Dichotomous data were analyzed by chi test; continuous data were analyzed by Student's t test. Any values of p < 0.05 were considered significant for any test. RESULTS: Both cohorts were comparable with regard to demographic data, ISS, AIS score in the thoracic region, and incidence and severity of brain injury. There was no difference in dependent parameters in both cohorts. Rates of ARDS, MOFS, and mortality were not negatively influenced by early unreamed IMN. CONCLUSION: Early unreamed IMN of femoral fractures in multiply injured patients with severe thoracic trauma is a safe procedure and seems to be justified to achieve early definitive care. PMID- 17414350 TI - Factors affecting length of stay after isolated femoral shaft fractures. AB - BACKGROUND: Controlling escalating health care costs is important for hospitals and has far reaching implications for society. Hospital length of stay (LOS) is one of the most reliable predictors of cost after trauma. Our purpose was to establish LOS after isolated femur fractures treated by intramedullary (IM) nail. METHODS: The trauma registry was queried to identify patients. Patients were excluded if they required assistive ambulatory devices before injury, had chronic/terminal illness or compromised brain function. There were 102 patients with an average age of 27 years. Statistical analysis was completed. RESULTS: The average LOS was 3.9 days. In all, 27 patients stayed longer than 4 days. Reasons included social (7), medical (10), and hospital delays (10). The average time from arrival to surgery was 17 hours. The average time from surgery to physical therapy was 1.3 days. Patients who had surgery more than 24 hours after arrival stayed longer. Patients requiring placement stayed 2.4 days longer until placement was found. CONCLUSIONS: Factors the hospital can control to reduce LOS include time to surgery, time for physical therapy evaluation, and radiology delays (for spine clearance radiographs). Physical therapy availability within 24 hours of surgery is important and should include weekends and holidays. Early evaluation of social factors including homelessness or obstacles to independent living may reduce time to find placement. LOS reduction after femur fractures will decrease the cost of trauma to the hospital. In addition, LOS reduction will possibly increase bed availability and minimize the time spent on diversion yielding greater revenues and increasing patient satisfaction. PMID- 17414351 TI - Edema in the lower limb of patients operated on for proximal femoral fractures. AB - BACKGROUND: Patients with proximal femoral fracture (PFF) often develop postoperative edema in the operated limb. This may lead to reduced mobilization, increasing the length of hospitalization. It is therefore relevant to gain information about the extent and pathogenesis of this edema formation. METHODS: Forty-one patients with PFF (30 women and 11 men) were studied pre- and postoperatively. Patients were grouped into pertrochanteric fractures and femoral cervical fractures, according to the AO/ASIF classification of PFF. Thigh and calf volumes were calculated in both fractured and contralateral limbs preoperatively and on postoperative days 3, 5, 7, and 30. RESULTS: All patients with PFF developed edema in the operated limb. The greatest volume increase occurred on postoperative day 7 (p < 0.0005). The magnitude of edema in the thigh and the leg of patients with pertrochanteric fractures as compared with the nonoperative side was approximately twice as great as in those with femoral cervical fractures (p < 0.0001). There was a statistically significant daily increase in the volume of the operative limb as compared with the nonoperative side. Age and sex were not correlated with the extent of edema formation. Functionally significant deep venous thrombosis and local infection could be excluded as causative factors. CONCLUSIONS: Postoperative edema in the thigh and leg of the operated limb was considerable. The magnitude of edema formation was related to the severity of primary trauma and the type of osteosynthesis. Therefore, the operation performed for PFF should be minimally traumatic. PMID- 17414352 TI - The identification of criteria to evaluate prehospital trauma care using the Delphi technique. AB - OBJECTIVE: Current trauma system performance improvement emphasizes hospital- and patient-based outcome measures such as mortality and morbidity, with little focus upon the processes of prehospital trauma care. Little data exist to suggest which prehospital criteria should serve as potential filters. This study identifies the most important filters for auditing prehospital trauma care using a Delphi technique to achieve consensus of expert opinion. METHODS: Experts in trauma care from the United States (n = 81) were asked to generate filters of potential utility in monitoring the prehospital aspect of the trauma system, and were then required to rank these questions in order of importance to identify those of greatest importance. RESULTS: Twenty-eight filters ranking in the highest tertile are proposed. The majority (54%) pertains to aspects of emergency medical services, which comprise 7 of the top 10 (70%) filters. Triage filters follow in priority ranking, comprising 29% of the final list. Filters concerning interfacility transfers and transportation ranked lowest. CONCLUSION: This study identifies audit filters representing the most important aspects of prehospital trauma care that merit continued evaluation and monitoring. A subsequent trial addressing the utility of these filters could potentially enhance the sensitivity of identifying deviations in prehospital care, standardize the performance improvement process, and translate into an improvement in patient care and outcome. PMID- 17414353 TI - Trauma education and care in the jungle of Ecuador, where there is no advanced trauma life support. AB - BACKGROUND: The advanced trauma life support course is not available or affordable to rural areas in low-income countries. A trauma continuing education course was created to educate physicians of rural hospitals in the jungles of Ecuador. METHODS: A basic trauma care course was designed based on local resources and location of injury, including rudimentary health posts in the jungle, rural hospitals, and definitive referral centers. Course effectiveness was evaluated by a comparison of test scores before and after the course. A multiple choice questionnaire was given. Comparison to previous test scores was also performed. Paired Student's t test was used for statistical analysis. An objective structured clinical examination (OSCE), based on the course design, was administered. RESULTS: Twenty-six rural physicians participated in the course. Mean test scores significantly improved from pretest to post-test (72% to 79%; p = 0.032). Knowledge deficiencies in prehospital care, extremity injury care, and patient evaluation adjuncts significantly improved from 23% to 87%, 23% to 100%, and 31% to 100%, respectively. Test results after the course showed improvements in all major categories tested. Twelve of the 26 participants were repeat test takers from a course provided 2 years earlier. These participants showed improved pretest scores compared with their highest previous test score (76.8% versus 68.5%; p = 0.0496). Compared with first-time test takers, these participants showed improved pretest (76.8% versus 68.4%) as well as post-test (81% versus 76%) scores. Twenty-five of the 26 physicians participated in the OSCE, with a pass rate of 76%. The OSCE identified various strengths and deficiencies based on patient location and available resources. In rudimentary health posts, management was adequate for hemorrhage control (65%), immobilization (77%), and early transfer to rural hospitals (92%). Prehospital communication was inadequate (53%). Rural hospital management was adequate for primary evaluation (60%) and resuscitation (74%) but poor in secondary patient evaluation (53%), adjuncts (25%), and transfer to definitive referral centers (11%). OSCE scores differed from multiple choice questionnaire test results. DISCUSSION: Where there is no advanced trauma life support, a tailored trauma course and evaluation can be effective in educating local providers. A well-designed competency evaluation (multiple choice questionnaire and OSCE) is helpful in identifying deficient local aspects of trauma care. The course design and evaluation methods may serve as a model for continuing trauma care education in developing countries. PMID- 17414354 TI - The interaction of 'occupant factors' on the lower extremity fractures in frontal collision of motor vehicle crashes based on a level I trauma center. AB - BACKGROUND: To advance occupant protection through the improvement of safety countermeasures, it is necessary to understand the factors that affect human injury tolerance. This study investigated the effect of 'occupant factors' i.e. age, gender, height, weight, and body mass index (BMI) on the pattern of lower extremity injury after motor vehicle crashes (MVCs). The second objective was to identify factor(s) (including restraint systems and Delta V) that influence the severity of fractures (open versus close fractures) within the lower extremity area. The outcome of this study may have implications toward secondary prevention in MVCs. METHODS: One-hundred and thirty-seven front-seat occupants involved in MVCs with frontal impact admitted to the University of Michigan trauma center as part of the Crash Injury Research and Engineering Network (CIREN) project were evaluated. Injuries were classified according to location (knee, thigh, hip [KTH]; lower leg [LL], foot and ankle [FA]). All the relevant variables mentioned above were analyzed. RESULTS: KTH fractures were the most common region (49.5%) affected, followed by fractures to the FA (38.4%) and LL region (12.1%). Female occupants, being generally shorter than their male counterpart, sustained a significantly higher percentage of FA fractures (44% vs. 29.5%, p < 0.05). Male occupants sustained more KTH fractures (58.3% vs. 44%, p < 0.05). Results demonstrated that there were significantly higher percentages of 'open' fractures in the below knee area (FA [53.8%], LL [24.4%], and KTH [21.8%]; p < 0.05). Of all those variables tested (age, gender, height, weight, BMI, restraint systems, and Delta V), occupant's height had a significant effect on the severity of fractures sustained. CONCLUSION: The interactive effect observed for height and gender on the pattern of lower extremity fracture is principally related to the body habitus and that gender may be a 'proxy' variable. The 'human factor' plays a vital role in influencing the pattern of injury in a MVC. This study strongly supports the fact that occupants with dissimilar body habitus interact differently with the interior cabin of the vehicle, thus, the performance of the active and passive safety systems. PMID- 17414355 TI - Firearm suicide: use of a firearm injury and death surveillance system. AB - BACKGROUND: Suicide is an important public health concern. Firearms are the most common mechanism of suicide death. This study describes the epidemiology of fatal and nonfatal firearm suicide injuries (FSI) in one metropolitan area from 2002 through 2004 using a firearm injury surveillance system. METHODS: Records were obtained of all victims of firearm injuries from hospitals, police, and the coroner. All injuries categorized as suicide were included. RESULTS: Local age adjusted suicide rates were significantly higher than state or national rates for ages 15 to 24, and significantly higher than national rates for ages 25 to 44. Men were FSI victims more than five times as often as women were. There was no seasonal pattern identified. Handguns were used nearly three out of four times. Eighty-six percent of FSI victims died, two-thirds at the scene. Most wounds were in the head or chest. Mental illness or relationship problems were common. Most suicides occurred in a residence. CONCLUSIONS: Community level firearm injury surveillance effectively identifies local trends that may differ from national statistics. Collaboration among various groups is used to support injury prevention programs. These data can both complement and contribute to national statistics. PMID- 17414356 TI - Equestrian injury prevention efforts need more attention to novice riders. AB - BACKGROUND: Equestrian injury is commonly seen at trauma centers and the severity of injury is often high. We sought to determine the risk, incidence, and the influence of skill and experience on injury during horse-related activity (HRA). METHODS: Members of horse clubs and individual equestrians in a three-state region (Oregon, Washington, and Idaho) were recruited via mailings and community advertisements to take a survey regarding their horse contact time and injuries over their entire riding career. Serious injury (SI) was defined by hospitalization, surgery, or long-term disability. RESULTS: There were 679 equestrians with a median age of 44 years who reported a median of 20 hours of HRA per month with a mean of 24 years (1 to 75 years) experience. The cumulative risk of any injury (AI) was 81% and of SI was 21%. The incidence of AI and SI were 1.6 +/- 0.1 (SE) and 0.26 +/- 0.02 per 10,000 hours, respectively. The incidence, per 10,000 hours, of AI was 7.6 +/- 2.7, 2.4 +/- 0.2, 1.5 +/- 0.1, and 1.0 +/- 0.1 at novice, intermediate, advanced, and professional levels, respectively (p < 0.001, analysis of variance [ANOVA]) and of SI was 1.03 +/- 0.52, 0.38 +/- 0.06, 0.21 +/- 0.03, and 0.19 +/- 0.04 at the respective skill levels (p < 0.001, ANOVA). There was a sharp decline in incidence of injury between 18 and 100 hours of experience. Helmet use was 74%, 61%, 58%, and 59% at the respective skill levels (NS, chi). CONCLUSION: One in five equestrians will be seriously injured during their riding career. Novice riders experienced a three-fold greater incidence of injury over intermediates, a five-fold greater incidence over advanced riders, and nearly eight-fold greater incidence over professional equestrians. Approximately 100 hours of experience are required to achieve a substantial decline in injury. These findings suggest that equestrian injury prevention efforts need more attention and should focus on novice equestrians. PMID- 17414357 TI - Contrast-enhanced CT accurately detects hemorrhage in torso trauma: direct comparison with angiography. AB - OBJECTIVE: To assess the ability of contrast enhanced computed tomography (CECT) to detect active hemorrhage and other vascular injuries in chest, abdominal, and pelvic trauma patients, using angiographic findings and need for intervention as paired gold standards. METHODS: We obtained approval from the Institutional review board for a retrospective search of the radiology information system: seeking trauma patients undergoing angiography within 24 hours of CECT for chest, abdominal, or pelvic injuries. CECT protocol was standard trauma CT, not specialized for CT angiography. Angiographic techniques varied with indication. Clinical and imaging reports and selected radiologic studies were reviewed, but the original dictated report was the interpretive standard. We used Fisher's exact test for statistical analysis. RESULTS: During the 30-month study period, 466 patients underwent emergent interventional radiologic procedures. Of those, 418 were excluded for nontrauma indications or neuroangiographic procedures. Fourty-eight patients (33 male, 15 female, average age 43.4) thus constituted the study population in whom we evaluated 63 traumatic injuries. CT findings had statistically significant associations (p < 0.0001) with both angiographic evidence of active hemorrhage and the need for intervention, which were tabulated separately. CT had 94.1% sensitivity and 97.6% negative predictive value (NPV) for detection of active hemorrhage, and 92.6% sensitivity and 91.2% NPV for predicting need for surgical or endovascular intervention. CONCLUSIONS: CECT findings correlate strongly with angiographic findings, though sensitivity remains imperfect. However, when CT is used in the context of other clinical features, particularly hemodynamic instability, it may enable clinicians to reserve emergent angiography for those patients in whom emergent intervention is planned. PMID- 17414358 TI - Fourteen-year review of open globe injuries in an urban Korean population. AB - OBJECTIVE: To review the epidemiology and prognostic factors related to visual outcome after open globe injuries. METHODS: A retrospective case review during a 14-year period. RESULTS: The study was based on a sample of 321 patients (321 eyes) with open globe injuries. Gender of the sample was predominantly male (76.6%) and the average age was 38.8 years. Injuries occurred most frequently in autumn (37.7%) and in the work place (45.8%). The anatomic site of the wound was corneal (zone I) in 148 eyes, corneoscleral (zone II) in 61 eyes, and scleral (zone III) in 92 eyes. Initial visual acuity was 4/200 or worse in 77.9% of patients and was significantly correlated with the zone of injury (linear by linear association p = 0.000). The elapsed time from injury to surgery was shorter than 48 hours in 90.7%. Primary repair was performed in 301 eyes and, subsequently, pars plana vitrectomy in 165 eyes, and cataract extraction in 67 eyes. Anatomic success was achieved in 91% of eyes and evisceration was performed in 7.8% (25 eyes). Final visual acuity of 5/200 or better was achieved in 58.3% of patients, and vision of 20/40 or better in 36.5%. The zone of injury was the most significant predictor of functional outcome (linear by linear association p = 0.000) as well as anatomic outcome (chi p = 0.000). Associated injuries such as retinal detachment, vitreous hemorrhage, uveal prolapse, and adnexal injury were also important prognostic factors (linear by linear association, p = 0.000). CONCLUSIONS: In this study, we identified the epidemiology and prognostic factors of open globe injuries in a Korean urban community. The visual as well as anatomic outcome for open globe injury was shown to be reasonably favorable when prompt and appropriate surgical management was applied. PMID- 17414359 TI - Wrist tourniquet: the most patient-friendly way of bloodless hand surgery. AB - BACKGROUND: The literature is scarce on wrist tourniquets. In this study, three well-established locations of tourniquet setting including upper arm, proximal forearm, and wrist were compared on the same limb using both clinical as well as biochemical variables in paramedical volunteers. METHODS: Twenty unmedicated, healthy, paramedical, right-hand dominant volunteers participated in the study. The left upper arms were used for monitoring. Blood pressures and heart rates were monitored and recorded before (baseline) and immediately after the application of the tourniquet, every 5 minutes, and at the time the patient requested deflation. An intravenous cannula (22 G) was placed on the right hand to obtain samples, which were taken at baseline and immediately after deflation of the tourniquet to evaluate the levels of pO2, pCO2, O2 saturation, pH, bicarbonate, blood sugar, lactate, hematocrit, and electrolytes. The tourniquets were applied to the right upper arm, forearm, and wrist of each subject with 5 day intervals between each trial. Subjective discomfort and tourniquet pain levels were recorded. For each trial, tourniquet tolerance and details of discomfort were recorded. Statistical analysis was performed as appropriate. RESULTS: Twenty volunteers aged 20 to 44 years were included. For each trial, in the first 10 minutes after inflation of the tourniquet, the heart rate and systolic blood pressure were increased compared with baseline values. Diastolic blood pressure was elevated immediately after inflation and remained so until deflation in each trial. Diastolic blood pressure values were higher in the upper arm tourniquet group compared with wrist. Then pH, pO2, and O2 saturation values were decreased and pCO2 and lactate levels were increased compared with baseline values in each trial. Blood sugar was decreased significantly in the arm group. The decrease in pH, pO2, O2 saturation, and blood sugar in the upper arm group was significantly higher compared with wrist and forearm groups. The lactate value was higher in the upper arm group compared with wrist. Visual analog scale and numerical rating scores were lower in the wrist group compared with others at all times. The longest tourniquet tolerance was in the wrist group. In the wrist group, curling was observed in all subjects but the fingers could easily be extended. CONCLUSION: The wrist tourniquet is the most comfortable technique of bloodless surgery for procedures limited to the hand region. PMID- 17414360 TI - Increased incidence of complications in trauma patients cointoxicated with alcohol and other drugs. AB - BACKGROUND: Alcohol and drug intoxication is prevalent in trauma patients. Although intoxication and cointoxication can have a range of physiologic effects, their implications for clinical management are unclear. The current investigation aims to assess the effects of alcohol and substance use as well as the interaction between these two states on outcomes and in-hospital complications. METHODS: All trauma patients with an Injury Severity Score (ISS) >or=12 during a 5-year period who were tested for both alcohol and other drugs were included. Alcohol-positive, drug-positive, and both-positive patients were compared with patients who tested negative. Logistic regression analysis was performed controlling for age and ISS to assess the relative contribution of intoxication or cointoxication in determining clinical outcomes and in-hospital complications. RESULTS: For alcohol-positive and drug-positive patients, intoxication status did not appear to influence outcomes. However, cointoxicated individuals were found to have an increased incidence of complications overall (odds ratio [OR] = 2.06), an increased incidence of pneumonia specifically (OR = 3.34) and an increased incidence of the requirement for mechanical ventilation (OR = 2.37). CONCLUSIONS: Cointoxication with alcohol and other drugs is a risk factor for increased in hospital complications. PMID- 17414361 TI - Two unusual cases of postamputation pain from Operation Iraqi Freedom. PMID- 17414362 TI - Small bowel injury from a tangential gunshot wound without peritoneal penetration: a case report. PMID- 17414363 TI - Emphysematous gastritis in a severely burned patient: case report and literature review. PMID- 17414364 TI - Ovarian tumor causing abdominal compartment syndrome. PMID- 17414365 TI - Pseudoaneurysm of superior mesentery artery branch after renal extracorporeal shock wave lithotripsy: case report and review. PMID- 17414366 TI - Severe accidental hypothermia successfully treated by rewarming strategy using continuous venovenous hemodiafiltration system. PMID- 17414367 TI - Decreasing the morbidity of decompressive craniectomy: the Tucci flap. PMID- 17414368 TI - Are diagnostic peritoneal lavage or focused abdominal sonography for trauma safe screening investigations for hemodynamically stable patients after blunt abdominal trauma? A review of the literature. AB - BACKGROUND: Assessment of patients in the emergency department who sustain blunt abdominal trauma represents a significant diagnostic challenge. Computed tomography (CT) is increasingly used as the principal investigation for these patients. A sensitive screening test could safely reduce the use of CT. OBJECTIVES: To appraise the evidence supporting the use of diagnostic peritoneal lavage and focused abdominal sonography for trauma as screening tests in the emergency department to reduce the use of CT in the initial assessment of patients sustaining blunt abdominal trauma. METHODS: A search of high-quality evidence resources was performed, followed by a hand search of the bibliographies of all relevant articles. RESULTS: Altogether, 55 articles were found during the initial search, of which 23 were relevant. An additional 11 were found by hand searching. Six relevant original research articles were found. CONCLUSION: Screening diagnostic peritoneal lavage and selective CT is a safe diagnostic strategy for the investigation of blunt abdominal trauma. Further research is needed to determine the role of focused abdominal sonography for trauma scanning in diagnostic protocols. PMID- 17414369 TI - Double dislocation of little finger in nonathletic person. PMID- 17414371 TI - Recognizing and responding to gallbladder woes. PMID- 17414370 TI - Abdominal trauma: dealing with the damage. PMID- 17414372 TI - Know the ABCs of caring for children. PMID- 17414375 TI - DepoDur extended-release epidural morphine: reshaping postoperative care. What perioperative nurses need to know. AB - Epidural morphine as a single bolus dose has demonstrated analgesia that lasts up to 24 hours. Recent advances in drug delivery mechanisms have resulted in a formulation of morphine, DepoDur, which is a lipid-encapsulated extended-release epidural morphine that provides up to 48 hours of analgesia. The efficacy of DepoDur has been established after hip arthroplasty, lower abdominal surgery involving an incision below the umbilicus, and elective cesarean section delivery. The unique characteristics of DepoDur dictate that clinicians must be aware of the benefits and risks, and facilities must have a comprehensive system to allow for the safe administration of DepoDur. This article provides information on this novel drug delivery system, reviews research findings reported in the literature, and describes the relational collaboration system designed and implemented at Duke University Health System for safe patient care of DepoDur recipients. PMID- 17414377 TI - African-American women: health beliefs, lifestyle, and osteoporosis. AB - Cultural beliefs of African-American women, manifested in lifestyle behaviors, may influence osteoporosis development. Genetics may have a positive influence on bone density, but the risks increase from other diseases that may contribute to the development of secondary osteoporosis. Health beliefs, lifestyle behaviors, and secondary osteoporosis causes are explored, focusing on relationships with osteoporosis. African-American women must be informed of the risks involved with some of the traditional health beliefs and practices. Healthcare providers have a challenge and responsibility to explore cultural beliefs with African-American women and to develop an acceptable approach to osteoporosis prevention. PMID- 17414379 TI - Spondylolysis: a review and treatment approach. AB - Low back pain, specifically "spondylo" conditions, has traditionally been misunderstood and often times ill-treated. A thorough understanding of the function of pars interarticularis and its relationship to the entire vertebral unit and low back health are essential for successful treatment and rehabilitation outcomes. Lifestyle awareness and controlled progression through the inflammation, stabilization, strength, and functional rehabilitation phases provide primary guidance for patients. In addition, a broad spectrum of pharmacological, psychological, therapeutic modality, and newer surgical techniques must be considered in the overall treatment plan. Having a strong understanding of the anatomy, biomechanics, treatment, and rehabilitation of this condition, will help the allied healthcare provider better meet the individualized needs of spondylolysis patients. PMID- 17414381 TI - Advancing evidence-based practice: a primer. AB - Evidence-based practice (EBP) requires a shift from the traditional paradigm of clinical practice grounded in intuition, clinical experience, and pathophysiological rationale. In the EBP paradigm, clinical expertise is combined with integration of best scientific evidence, patient values and preferences, and the clinical circumstances. This primer article provides a summary of driving forces mandating EBP, barriers to EBP, and an overview of the EBP process highlighting critical resources and practices. PMID- 17414383 TI - Changing paediatric orthopaedic education and certification for the RN in Northern Ireland? AB - This article highlights the development of orthopaedic nurse education in Northern Ireland and how it equates with the United Kingdom and United States when focused on children's nursing. It also examines the historical development of initial preregistration nurse education and equates it to children's nursing where children are generally acknowledged as a vulnerable group. This article concludes by offering recommendations that may further enhance the development of Northern Ireland's orthopaedic nurse education that demonstrate standardization and equality for all practicing nurses within the specialty of orthopaedics. PMID- 17414384 TI - Patient safety, error reduction, and ethical practice. AB - Currently, patient safety and adverse outcomes are receiving an increasing emphasis within both the healthcare community and society. Redesigning systems and instituting safe practices within the healthcare environment are being seen as essential to decrease the likelihood that patients are placed at risk of harm. Adopting a change in practice may not be the answer because there may be unintended consequences from the new procedure. The purposes of this article are to briefly describe the phenomena of patient safety and error reduction, identify corresponding ethical concerns, and discuss selected implications related to patient safety that promote ethical practice. PMID- 17414385 TI - Medicating young or very young patients--part I. AB - Children enter an orthopaedic treatment environment for a variety of reasons. Some of these pediatric patients will be seen in practices specializing in pediatric orthopedics. However, many young patients with less demanding problems will be seen in family practice settings and general orthopaedic areas. Medicating these children safely and effectively and educating parents about a child's medications are important aspects of good nursing care. In this first discussion about pediatric medications, the unique considerations about medicating children at different ages are addressed. Adherence considerations are identified, guidelines for talking with children about medications are identified, and improvements in the regulations that are designed to increase knowledge about safety and efficacy in pediatric pharmacology are reviewed. The second part of this discussion addresses examples of drugs used to address selected common health problems in pediatric patients and will be published in the May/June 2007 issue of Orthopaedic Nursing. PMID- 17414387 TI - Pain in the orthopaedic patient. PMID- 17414388 TI - Natural history of plasma leakage in dengue hemorrhagic fever: a serial ultrasonographic study. AB - BACKGROUND: Although plasma leakage is the major cause of mortality and morbidity in patients with dengue hemorrhagic fever (DHF), a detailed assessment of the natural course of this process is still lacking. We employed serial ultrasound examination to delineate the locations and the timing of plasma leakage and to evaluate the usefulness of ultrasound in detecting plasma leakage in DHF. METHOD: Daily ultrasound examinations of the abdomen and right thorax were performed in 158 suspected dengue cases to detect ascites, thickened gall bladder wall and pleural effusions. Cases were classified into dengue fever (DF), DHF or other febrile illness (OFI) based on serology and evidence of plasma leakage including hemoconcentration and pleural effusion detected by chest radiograph. RESULTS: Ultrasonographic evidence of plasma leakage was detected in DHF cases starting from 2 days before defervescence and was detected in some cases within 3 days after fever onset. Pleural effusion was the most common ultrasonographic sign of plasma leakage (62% of DHF cases one day after defervescence). Thickening of the gallbladder wall and ascites were detected less frequently (43% and 52% of DHF cases respectively) and resolved more rapidly than pleural effusions. The size of pleural effusions, ascites and gall bladder wall thickness in DHF grade I and II were smaller than those of grade III patients. Ultrasound detected plasma leakage in 12 of 17 DHF cases who did not meet the criteria for significant hemoconcentration. CONCLUSIONS: Ultrasound examinations detected plasma leakage in multiple body compartments around the time of defervescence. Ultrasonographic signs of plasma leakage were detectable before changes in hematocrits. Ultrasound is a useful tool for detecting plasma leakage in dengue infection. PMID- 17414390 TI - Transmission of Bordetella pertussis to young infants. AB - BACKGROUND: Pertussis vaccination has reduced the number of notified cases in industrialized countries from peak years by more than 95%. The effect of recently recommended adult and adolescent vaccination strategies on infant pertussis depends, in part, on the proportion of infants infected by adults and adolescents. This proportion, however, remains unclear, because studies have not been able to determine the source case for 47%-60% of infant cases. METHODS: A prospective international multicenter study was conducted of laboratory confirmed infant pertussis cases (aged or=2) were significantly less in the zinc group. Approximately 85% of the surveillance workers made weekly visits to the houses and 96% of mothers administered syrup weekly to their children. Around 80% of mother's were aware of the possible benefits of zinc supplementation. CONCLUSION: Weekly zinc supplementation was effective in reducing diarrheal morbidity at the community level and it was operationally feasible. PMID- 17414393 TI - Lack of usefulness of an abnormal white blood cell count for predicting a concurrent serious bacterial infection in infants and young children hospitalized with respiratory syncytial virus lower respiratory tract infection. AB - BACKGROUND: There continues to be controversy on the most appropriate way to manage infants and young children with fever and documented RSV lower respiratory tract infection (LRTI). The objective of this study was to determine the usefulness of an abnormal white blood cell (WBC) count for predicting a concurrent serious bacterial infection in patients admitted with RSV LRTI. METHODS: The medical records were reviewed of patients discharged with RSV LRTI during the 5 RSV seasons from July 1, 2000 through June 30, 2005. Data were collected on age and gender as well as temperature, complete blood count with manual differential and bacterial cultures obtained at admission. RESULTS: The inclusion criteria was met by 1920 patients. There were 672 febrile patients who had a complete blood count and a bacterial culture. One (5.0%) of 20 patients with a WBC <5000 had a positive culture, 23 (4.7%) of 492 patients with a WBC 5000-14,999 had a positive culture, 5 (4.8%) of 105 patients with a WBC 15,000 19,999 had a positive culture, 2 (5.7%) of 35 patients with a WBC 20,000-24,999 had a positive culture, none of 11 patients with a WBC 25,000-29,999 had a positive culture and 3 (33%) of 9 patients with a WBC >30,000 had a positive culture. Overall, cultures were positive in 34 (5.1%; 95% CI: 3.4-6.8%) of the febrile patients tested and almost all (32; 94%) showed positive urine cultures. CONCLUSION: The probability of an abnormal WBC count <5000 and 15,000-30,000 being associated with a concurrent serious bacterial infection was very low and no different from that of a normal WBC count in febrile patients admitted with RSV LRTI. PMID- 17414394 TI - Pertussis is common in nonvaccinated infants hospitalized for respiratory syncytial virus infection. AB - BACKGROUND: There is some evidence that Bordetella pertussis can cause co infection with respiratory syncytial virus (RSV). SUBJECTS AND METHODS: RSV etiology was studied by antigen detection in 117 infants <6 months of age, who were hospitalized for respiratory tract infection during an RSV epidemic. B. pertussis etiology was studied by polymerase chain reaction in those 88 in whom parents or nurses reported cough. RESULTS: RSV was found in 91 (78%) infants and B. pertussis in 9 (8%) infants. In 7 cases, there was mixed RSV-pertussis infection. In retrospective analysis, RSV and mixed RSV-pertussis cases could not be separated by clinical characteristics. CONCLUSIONS: Co-infection caused by B. pertussis was present in 8% of infants, aged <6 months, who were hospitalized for RSV infection. To avoid under-diagnosis, pertussis should be considered in all nonvaccinated infants with lower respiratory tract infection. PMID- 17414395 TI - Longitudinal study on pneumococcal carriage during the first year of life in Bangladesh. AB - BACKGROUND: The strong herd immunity effect and the serotype replacement associated with the use of the pneumococcal conjugate vaccine have highlighted the importance of asymptomatic pneumococcal carriage. To describe the development of pneumoccoccal carriage in a developing country setting we carried out a longitudinal pneumococcal carriage study in Bangladesh. METHODS: Ninety-nine children, born in Savar, Bangladesh between May 2000 and April 2001, were enrolled in the study with their families. Nasopharyngeal samples were collected at prescheduled 2-4 week intervals from the index children and from their family members. The nasopharyngeal swabs were cultured for pneumococcal growth and pneumococci were identified and serotyped by standard methods. RESULTS: We collected 1459 samples (92% of those planned) from the 99 index children and 2865 samples from other family members. The data showed high point prevalences of pneumococcal carriage among newborns (40-50% from 8 weeks of age on), a rapid pneumococcal acquisition with age (50% of the children had been colonized by pneumococci at least once by the age of 8 weeks) and a wide range of different serogroups/types (SGT). SGT 6 and 19 accounted for 35% of the pneumococci isolated from children <1-year-old, followed by SGT 15, 23, and 10 for a total of 56%. The SGT distribution in children up to 9-year-old was similar to that among the <1 year olds, with SGT 6 and 19 predominating. Older children and adults differed from the younger children by not having clearly predominating SGTs. CONCLUSIONS: The features found in our study are typical of pneumococcal carriage in developing countries. We believe that results from longitudinal modeling of carriage based on these extensive data can have wide geographic application. PMID- 17414396 TI - Rotavirus disease in young children from Hanoi, Vietnam. AB - BACKGROUND: Pathogen prevalences and antimicrobial susceptibilities are essential for the rational development of preventive strategies for diarrheal diseases, but little recent information from Vietnam is available. We prospectively studied the prevalence of enteric pathogens in children less than 5 years of age with acute diarrhea and in nondiarrhea controls in a city hospital in Hanoi, Vietnam for 1 year. METHODS: Enteric bacteria and viruses were detected by standard culture methods, and enzyme immunoassay in 291 cases and 291 controls. RESULTS: Detection rates of viral pathogens among cases and controls were 31% and 3% for rotavirus, 12% and 1% for astrovirus and 4% and 1% for adenovirus. For bacterial pathogens, Aeromonas, Shigella, Salmonella, Campylobacter and enterotoxigenic E. coli were isolated from cases and controls in 15% and 8%, 9% and 1%, 7% and 1%, 4% and 0%, and 3% and 0%, respectively. The isolation of bacterial and viral pathogens except for adenovirus was significantly lower in controls than cases. Fluoroquinolones were effective against most bacterial enteropathogens, but resistance was observed in 27% of Campylobacter isolates. CONCLUSIONS: Viral etiologic agents especially rotavirus were the most important cause of acute diarrhea in children less than 5 years of age in Hanoi. The burden of rotavirus in young children in Hanoi warrants consideration of using the recently released rotavirus vaccine. PMID- 17414397 TI - Perinatal hepatitis B transmission and vaccination timing in a managed care cohort: assessment of the temporary delay in newborn hepatitis B vaccination due to thimerosal content. AB - BACKGROUND: From July to September 1999, due to a concern of toxicity from exposure to thimerosal-containing vaccines, the American Academy of Pediatrics and U.S. Public Health Service temporarily recommended delaying the administration of first dose of hepatitis B vaccine until the age of 2-6 months for infants born to hepatitis B surface antigen negative mothers. Our objectives were to determine whether the recommendation affected the rate of perinatal hepatitis B infection in a multistate managed care population; to describe neonatal and early childhood cases of hepatitis B infection and to evaluate a possible role of the recommendation; and to assess the timeliness, with respect to the U.S. childhood immunization schedule, of vaccinations during the first 2 years of life. METHODS: We identified 3 cohorts of infants born before (July 1998 to June 1999), during (July 1999 to September 1999) and after (October 1999 to September 2000) the recommendation period. We used automated claims data to identify possible neonatal and early childhood hepatitis B cases using specific ICD-9 diagnosis and CPT procedure codes and validated cases through medical record review. Using Health Plan Employer Data and Information Set (HEDIS) data, we calculated vaccination coverage for the first dose of hepatitis B vaccine at 3 month intervals from January 1999 to September 2000. RESULTS: The eligible populations in the "before," "during" and "after" cohorts were 29,347, 7791 and 29,215 infants, respectively. Of 41 possible hepatitis B cases identified in the 3 cohorts, we confirmed 1 case in the after cohort with medical record review. Despite receiving the first dose of hepatitis B vaccine and hepatitis B immunoglobulin within 12-24 hours of birth, the infant was diagnosed with laboratory-confirmed chronic hepatitis B at age of 9 months. An analysis of HEDIS data showed that vaccination coverage for the first dose of hepatitis B vaccine was 98% (January to March 1999) and 96% (April to June 1999) for the "before" cohort and 66% for the "during" cohort. For the "after" cohort the coverage was 72% (October to December 1999), 83% (January to March 2000), 91% (April to June 2000) and 95% (July to September 2000). CONCLUSIONS: This study did not identify any perinatal hepatitis B transmission among health plan enrollees associated with the 1999 recommendation. The recommendation did result in a delay of hepatitis B birth dose in the "during" cohort as intended for infants born to hepatitis B surface antigen negative mothers. Six months after the recommendation was rescinded there was still a delay in the timing of first dose of hepatitis B vaccine, but the timing had returned to the prerecommendation level after 9-12 months. PMID- 17414398 TI - Impact on weight and height with the use of HAART in HIV-infected children. AB - BACKGROUND: There are few data on long-term effects of highly active antiretroviral therapy (HAART) on weight and height in HIV-infected children. OBJECTIVES: Our objective was to assess the effect of HAART on the weight and height of HIV-infected children over time in the Madrid cohort, and analyze possible factors associated with the effect. PATIENTS AND METHODS: This was a retrospective study of HIV-infected children starting HAART in 1997 or later. Serial measurements of weight, height and body mass index (BMI) were performed and converted to z-scores using the Spanish revised reference data. Changes from baseline in weight, height and BMI at 12, 24, 36, 48 and 60 months were determined. Associations of z-scores at the last visit with immunologic (CD4% above 25%) and virologic responses (more than 50% of samples below 400 copies/mL), CDC (Centers for Disease Control) clinical category, and the presence and type of lipodystrophy (lipoatrophy or lipohypertrophy) were evaluated. RESULTS: Twelve hundred and twelve children, 97% of them vertically-infected, received HAART starting in 1997 for a median of 71 months (4-102 months). Median age at initiation of HAART was 6 years (1 month-18 years). Thirty-nine percentage were antiretroviral naive and 61% had received NRTI therapy previously. Thirty two percentage and 53% had CDC class C and immunologic class 3, respectively. At the final evaluation, 24% of children remained on their first combination therapy, 39% on the second and 37% had received at least 3 different HAART regimens. Fifty-one percentage were classified as virologic responders. Thirty nine percentage of children in this study were diagnosed with lipodystrophy. At baseline, median z-score for weight, height and BMI were -0.45, -0.60 and -0.33, respectively. HAART was associated with significant increases in z-scores of weight and height but not BMI at the different time-points analyzed. Virologic nonresponders had significantly lower z-scores for weight and height but not for BMI. CDC class C was associated with lower z-scores for height. No differences in final measurements were observed for baseline CD4, immunologic response or lipoatrophy. Children with lipohypertophy had a significantly higher BMI at the last visit. CONCLUSIONS: HIV-infected children experienced a continued catch-up in weight and height 5 years after starting HAART. Virologic control is related to sustained growth. PMID- 17414399 TI - Epidemiology of methicillin-resistant Staphylococcus aureus at a pediatric healthcare system, 1991-2003. AB - BACKGROUND: The emergence and epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) at a Minneapolis pediatric healthcare facility was investigated. METHODS: Children with MRSA infections from January 1991 to December 2003 were classified as community-associated (CA) or healthcare associated (HA) using established criteria. Isolates were subtyped using pulsed field gel electrophoresis and grouped into pulsed-field types (PFTs). Case and isolate characteristics were compared and temporal trends were assessed. RESULTS: The first isolate classified as CA-MRSA in this healthcare facility was identified in 1991. CA-MRSA cases (n = 188) were more likely than HA-MRSA cases (n = 83) to have a skin or soft tissue infection (80% versus 59%) and to belong to a racial or ethnic minority group (82% versus 55%), whereas HA-MRSA cases were younger (median age, 3.4 years versus 4.9 years). The proportion of both CA- and HA-MRSA isolates susceptible to clindamycin and erythromycin declined during the study period. Isolates classified as CA-MRSA were more likely than HA-MRSA isolates to be USA300 (21% versus 11%, P = 0.05) and USA400 (62% versus 31%, P < 0.001) PFTs. Associations between case race/ethnicity and isolate PFT were observed independent of case classification. CONCLUSIONS: CA-MRSA is well established in this pediatric population. Although no discernable changes in CA- or HA-MRSA case characteristics were documented during the study period, significant changes were observed in CA-MRSA isolate characteristics, indicating that this pathogen continues to evolve. PMID- 17414400 TI - New zealand epidemic strain meningococcal B outer membrane vesicle vaccine in children aged 16-24 months. AB - BACKGROUND: New Zealand has experienced an epidemic of Neisseria meningitidis dominated by strain B:4:P1.7b,4 since 1991. Children younger than 5 years are at highest risk. Previous serogroup B outer membrane vesicle (OMV) strain specific vaccines have shown variable efficacy in this age group. OBJECTIVE: To evaluate the immunogenicity, reactogenicity and safety in 16-24-month-old children of an OMV vaccine developed against the New Zealand epidemic strain. METHODS: Children (332) aged 16-24 months were randomized to receive the New Zealand candidate vaccine made using strain NZ98/254 (B:4:P1.7b,4) or the Norwegian parent vaccine made using strain 44/76 (B:15:P1.7,16). Vaccines (25 microg/dose) were administered at 0, 6 and 12 weeks in this observer-blind trial. Immune response was measured by serum bactericidal assay and enzyme-linked immunosorbent assay. Sero-response was defined as a 4-fold or greater rise in serum bactericidal antibody titer compared with baseline, with titers <1:4 required to increase to >or=1:8 to be considered a sero-response. Local and systemic reactions were monitored for 7 days after vaccination. RESULTS: Sero-response against NZ98/254 was achieved after 3 doses in 75% (95% CI: 69-80%) receiving the New Zealand candidate vaccine by both intention to treat (ITT) and per protocol (PP) analyses. In Norwegian parent vaccinees this was seen in 3% (0-12%) (ITT) and 4% (0-13%) (PP). Vaccines were well tolerated with no vaccine-related serious adverse events. CONCLUSION: The New Zealand candidate vaccine administered to these 16-24-month-old children in 3 doses was safe and elicited a promising immune response against the candidate vaccine strain NZ98/254 (N. meningitidis B:4:P1.7b,4) contributing to vaccine licensure for this age group. PMID- 17414401 TI - Rabies-update on a global disease. PMID- 17414402 TI - The hereditary autoinflammatory syndromes. PMID- 17414403 TI - Active tuberculosis among adolescents in toronto, Canada: clinical features and delays in diagnosis. AB - There are few studies on tuberculosis (TB) in adolescents. During a 6-year period, 23 adolescents were treated for TB in our center. Most were foreign born. The average time from symptom onset to final diagnosis was 5.2 months. Multifocal and extra pulmonary disease occurred in the majority of patients. Clinicians need greater awareness of the unique features of adolescent TB. PMID- 17414404 TI - Introduction of Haemophilus influenzae type B conjugate vaccine into routine immunization in Ghana and its impact on bacterial meningitis in children younger than five years. AB - This report shows the impact of a pentavalent vaccine that includes Haemophilus influenzae type b (Hib) conjugate vaccine on bacterial meningitis in children younger than 5 years in Ghana. A review of the first 3 years of a pediatric bacterial meningitis surveillance program, started in August 2001 in Accra, Ghana, was undertaken. There was a significant reduction, P = 0.042 and 0.017, in percentage of purulent meningitis in children younger than 1 year, comparing the first year when the vaccine was introduced, to the second and third years, respectively. PMID- 17414405 TI - Opsoclonus-myoclonus syndrome associated with group a streptococcal infection. AB - This report describes the unusual syndrome of opsoclonus-myoclonus associated with group A streptococcal infection in a 9-year-old Nepalese boy. PMID- 17414406 TI - Effectiveness of early initiation of protease inhibitor-sparing antiretroviral regimen in human immunodeficiency virus-1 vertically infected infants. AB - Each of the 17 vertically infected infants born to HIV-1-infected mothers in Belgian HIV reference centers since 1996 was treated with a combination of 3 reverse transcription inhibitors as soon as the diagnosis was established. Treatment was initiated in all patients before 66 days of life. Twelve patients, including 11/13 infants treated with the combination of zidovudine, lamivudine and nevirapine, experienced a complete viral suppression (<50 copies/mL) with their first drug regimen. At last follow-up, 12 patients were asymptomatic, 2 were CDC stage A and 3 were stage B; 15 had HIV-1 RNA levels of <50 copies/mL and 14 had >or=25% CD4 lymphocytes. These results suggest that early initiation of treatment with 3 reverse transcription inhibitors is highly effective to inhibit viral replication and to prevent clinical and immunologic progression of HIV infection in vertically infected infants. PMID- 17414407 TI - Mycobacterium simiae cervical lymphadenitis. AB - Mycobacterium simiae, a multidrug-resistant, opportunistic acid-fast bacillus, usually causes infection in immunocompromised hosts. We describe a previously healthy child with M. simiae necrotizing granulomatous cervical lymphadenitis. Cure was achieved with excision of the affected nodes and adjunctive antimicrobial therapy. PMID- 17414408 TI - Successful medical treatment of cutaneous aspergillosis in a premature infant using liposomal amphotericin B, voriconazole and micafungin. AB - Treatment options for primary cutaneous aspergillosis in neonates are limited by the lack of pharmacokinetic and safety data of newer antifungal agents that are effective against Aspergillus spp. We report the successful treatment of cutaneous aspergillosis in an extremely low-birth-weight preterm infant with liposomal amphotericin B, voriconazole and micafungin, and provide pharmacokinetic profiles for voriconazole and micafungin. PMID- 17414409 TI - A case of interleukin-12 receptor beta-1 deficiency with recurrent leishmaniasis. AB - Interleukin-12 receptor beta-1 (IL-12Rbeta1) defect is generally associated with selective susceptibility to weakly pathogenic mycobacteria and Salmonella species. Patients rarely experience infections caused by other organisms. We report a 5-year-old patient with IL-12Rbeta1 deficiency who developed recurrent visceral leishmaniasis 6 months apart. The patient responded to lyposomal amphotericin B treatment reasonably well. PMID- 17414410 TI - Monoarthritis in rheumatic fever? PMID- 17414411 TI - Postoperative gram-negative anaerobic bacterial endocarditis. PMID- 17414412 TI - Vaccination usage among an old-order Amish community in Illinois. PMID- 17414414 TI - Dynamic near-infrared spectroscopy measurements in patients with severe sepsis. AB - This study evaluated near-infrared spectroscopy (NIRS)-derived measurements in hemodynamically stable patients with severe sepsis, as compared with similar measurements in healthy age-matched volunteers. Prospective, preliminary, observational study in a surgical intensive care unit and clinical research center at a university health center. We enrolled 10 patients with severe sepsis and 9 healthy age-matched volunteers. For patients with severe sepsis, we obtained pulmonary artery catheter and laboratory values three times daily for 3 days and oxygen consumption values via metabolic cart once daily for 3 days. For healthy volunteers, we obtained all noninvasive measurements during a single session. We found lower values in patients with severe sepsis (versus healthy volunteers), in tissue oxygen saturation (StO2), in the StO2 recovery slope, in the tissue hemoglobin index, and in the total tissue hemoglobin increase on venous occlusion. Patients with severe sepsis had longer StO2 recovery times and lower NIRS-derived local oxygen consumption values versus healthy volunteers. In our preliminary study, NIRS provides a noninvasive continuous method to evaluate peripheral tissue oxygen metabolism in hemodynamically stable patients with severe sepsis. Further research is needed to demonstrate whether these values apply to broader populations of patients with systemic inflammatory response syndrome, sepsis, severe sepsis, and septic shock. PMID- 17414415 TI - Is there a self-preserving hypothermic mechanism in shock? AB - Hypoxia-induced hypothermia (HIH) is regarded as an adaptive response to hypoxia in a variety of creatures, but no details of the mechanism have yet been elucidated in the clinical setting. This study was designed to analyze alteration of core body temperature with hemorrhagic shock and to clarify HIH in the clinical setting. Patients were categorized in the hemorrhage shock (S, n = 15) or cardiopulmonary arrest (C, n = 88) group. The tympanic membrane temperature (TMT) was measured, and the length of the interval of call-to-arrival (CTA) at a hospital was set as the time-course parameter. There was a significant negative linear relationship between CTA interval and TMT (S group: TMT = -0.055 degrees C, CTA = +36.1 min, r = -0.833, P < 0.001; C group: TMT = -0.046 degrees C, CTA = +36.3 min, r = -0.548, P < 0.001). Analysis of variance revealed no significant difference in the slope of the regression lines of both groups. However, when the CTA interval was used as a covariate, there was a significant difference in the TMT (P = 0.014), which means that the regression line of the S group was significantly lower than that of the C group with time. Furthermore, in the S group, all patients were hypothermic (<35 degrees C) when their CTA interval was more than 20 min; on the other hand, in the C group, only 64 (75%) of 85 were hypothermic. Patients in S group were more likely to become hypothermic (P < 0.05). In humans with cellular hypoxia, HIH takes place, as seen in other animals. This result emphasizes the necessity for studies of analysis of the mechanisms of temperature control and determination of optimal body temperature during acute critical care. PMID- 17414416 TI - A role for epinephrine in post-traumatic hypokalemia. AB - We have previously described a high incidence of admission hypokalemia in trauma patients at our institution. We subsequently performed a prospective study of 112 trauma patients to examine the possible etiologies of post-traumatic hypokalemia. Trauma patients >or=5 years old were evaluated within 6 h of injury with a variety of studies including catecholamines, cortisol, and insulin levels, with studies repeated 24 to 36 h after admission. No potassium replacement was given during this time. Demographic factors such as age, types of injury, and severity of injuries were collected. We found that the mean age of those with post traumatic hypokalemia (2 MF/50 high-power fields have shown potential for aggressive behavior including metastasis. OFMT with these features should be considered sarcomas. Given the histologic features, this tumor was considered a malignant OFMT. The patient had a wide excision. The patient died secondary to unrelated comorbidities without evidence of recurrence or metastasis. PMID- 17414438 TI - Unusual benign myxoid nerve sheath lesion: myxoid palisaded encapsulated neuroma (PEN) or nerve sheath myxoma with PEN/PEN-like features? AB - A 38-year-old woman had a polypoid nodule on her scalp. The lesion histopathologically demonstrated an extensive myxoid lobular lesion associated with a nonmyxoid and cellular area in the peripheral area of the lesion. The features of the peripheral cellular area resembled those of palisaded encapsulated neuroma (PEN). Most of the lesion was myxoid, with a large lobule in which several thin fibrous septa were present, dividing it into smaller lobules. The myxomatous area was composed of spindle cells or stellate cells with elongated cytoplasmic processes. There was a loose cellular network with a reticular and netlike network pattern or a lamellar pattern. From the immunohistochemical findings, as well as the histopathologic features, the whole lesion was thus considered to be a PEN, whereas the main, myxoid, lobular lesion was thought to have been caused by myxoid changes within a PEN, namely, myxoid PEN. It cannot be completely ruled out, however, that this case could be that of a nerve sheath myxoma with a PEN/PEN-like lesion. Thus, the present case may suggest that PEN can show extensive myxoid change or may demonstrate a relationship between PEN and nerve sheath myxoma. PMID- 17414439 TI - Cutaneous intralymphatic histiocytosis associated with rheumatoid arthritis: report of a case and review of the literature. AB - Various dermatoses have been described associated with rheumatoid arthritis. Recently, a specific cutaneous lesion termed "intravascular histiocytosis" has been proposed as a new entity among these dermatoses. We report the case of a 50 year-old woman with rheumatoid arthritis for about 10 years who developed erythematous patches on the extensor surface of lower extremities. Histopathologically, the lesions showed intraluminal proliferation of CD68 positive histiocytes in vessels lined with endothelial cells expressing D2-40, a selective marker for lymphatic endothelium. PMID- 17414440 TI - A metastatic melanoma with an unusual immunophenotypic profile. AB - The most commonly used melanocytic markers are S100, HMB45, Melan-A, or MART-1 and tyrosinase. Melanoma with complete, concordant loss of these markers has not been reported. We report a case of metastatic melanoma with complete loss of staining for S100, HMB45, Melan-A, and tyrosinase. Interestingly, both the primary melanoma and its metastasis were strongly positive for CD99. PMID- 17414441 TI - Scrotal calcinosis results from calcification of cysts derived from hair follicles: a series of 20 cases evaluating the spectrum of changes resulting in scrotal calcinosis. AB - Scrotal calcinosis is a rare disorder characterized by multiple papules or nodules of calcification in the scrotal skin. The etiology of this entity is speculative largely as a result of the paucity of larger series.This study of 20 patients with scrotal calcinosis was undertaken to critically analyze the histology with a view to assess the probable etiology of this lesion. Two thirds of the patients were young adults. Of these, 11 patients (55%) were asymptomatic and 9 complained of symptoms related to the breakdown of these lesions (eg, discharge, itching, or heaviness in the scrotum). All cases showed classical histologic features of scrotal calcinosis with a variable amount of calcification in the dermis. In 14 cases the etiology of this calcification could be traced to originate from dilated epidermal cysts. The spectrum of changes probably started with the cystic dilation of the hair follicle, then calcification around and within this cyst. Finally the epithelial elements disappeared, leaving behind residual areas of calcification. The remaining 6 cases did not have epithelial cysts in the vicinity. Based on these observations we conclude that scrotal calcinosis results from calcification of hair follicular or epidermal cysts, but as most of the cases report, this epithelium disappears and may not be seen. PMID- 17414442 TI - Eccrine porocarcinoma: a case with an obscure primary tumor diagnosed from lymph node metastasis. AB - Eccrine porocarcinoma (EPC), a rare malignant epidermal appendage tumor, is mainly seen in elderly patients. A long history is one of its main characteristics. Two types of EPC are known: juxtaepidermal and dermal. The juxtaepidermal type usually has a more aggressive behavior. Lymph node metastasis and high mitotic activity are associated with poor prognosis. A case of EPC with a long-standing but indolent primary site diagnosed from lymph node metastasis is presented here. There was also accompanying chronic dermatitis as a paraneoplastic syndrome. Diagnosing EPC prior to the regional lymph node involvement is the most valuable factor for a successful treatment. Persistent examinations and attempts to find the primary site(s) have to be made in such cases. PMID- 17414443 TI - Merkel cell tumor in a trichilemmal cyst: collision or association? AB - An 86-year-old white male presented with an erythematous, painless, slowly growing, and firm left thigh nodule. Histologic examination revealed a dermal proliferation of monomorphous cells arranged in trabeculae, nests, and sheets with an infiltrative growth pattern. The cells had a high nuclear-cytoplasmic ratio, finely granular nuclear chromatin, and nuclear molding. Numerous mitotic figures, apoptotic cells, and individual cell necrosis were present; lymphovascular invasion was identified. The tumor was attached, demonstrating pagetoid intraepithelial migration, to a follicular cyst lined by squamous epithelium, lacking a granular cell layer and filled with compact keratinous content, diagnostic of trichilemmal cyst. Immunohistochemical study revealed that tumor cells expressed pan-cytokeratin (CK), chromogranin, synaptophysin, neuron specific enolase, and CK20 (dotlike staining pattern), thus supporting the diagnosis of Merkel cell carcinoma. The association of Merkel cell carcinoma with a cyst is an exceptionally rare occurrence. As a result of the prominent involvement of the cyst wall by tumor cells, we favor that in this case carcinoma arose in the trichilemmal cyst rather than being a collision tumor. This hypothesis is also supported by the recent observation that Merkel cells are frequently present within normal hair follicles, especially in the isthmic portion that corresponds with the area of origin of the trichilemmal cyst. PMID- 17414444 TI - Cutaneous involvement with Burkitt-like lymphoma. AB - We report a case of a 27-year-old woman with a known history of Burkitt-like lymphoma (BLL), who presented with a rapidly enlarging nodule in the suprapubic area. Skin biopsy of the suprapubic nodule was consistent with cutaneous involvement of BLL. BLL is a highly aggressive B-cell lymphoma with a high proliferative rate. Like Burkitt lymphoma, BLL is characterized by a translocation of the c-MYC proto-oncogene. Histopathologic characteristics are considered borderline between those of classic Burkitt lymphoma and diffuse large B-cell lymphoma. Cutaneous involvement of BLL is rare and poorly documented in the literature. PMID- 17414445 TI - Abundant eosinophil infiltration in a neurofibroma. AB - We report a case of sporadic cutaneous neurofibroma showing diffuse eosinophil infiltration in a 72-year-old woman. The neurofibroma had been on the skin of the left anterior shoulder for approximately 10 years. The presence of mast cells is well known in neurofibroma. However, infiltration of neurofibromas by eosinophils has not yet been documented. We propose that eosinophil infiltration is mediated by the effects of mast cells. We suggest that this is indirect evidence that mast cells in some neurofibromas are not quiescent but active. We also reintroduce the notion that acid-fast stains are a useful method to discriminate between mast cell granules and eosinophil granules. PMID- 17414446 TI - Cutaneous epithelioid angioleiomyoma with clear-cell change. AB - Epithelioid angioleiomyoma is rare in the skin and subcutis. We report here an unusual case of epithelioid angioleiomyoma with prominent clear-cell change. Smooth-muscle differentiation was confirmed by immunostains. The extensive clear cell change in epithelial cells and the marked hyalinization and calcification made the histologic differential diagnosis challenging. Epithelioid angioleiomyoma shares some histologic characteristics with glomus tumors and seems to be an intermediate entity between angioleiomyomas and glomus tumors. PMID- 17414447 TI - Unique histopathologic findings in a patient with adult-onset Still disease. AB - Adult-onset Still disease (AOSD) is an uncommon disorder characterized by fever, polyarthralgia, elevated white blood cell count, and a maculopapular rash, the histologic features of which have not been well-known. A 55-year-old Asian woman presented initially with a "burning" and severely pruritic eruption on her face, hands, and arms, thought clinically to be urticaria. Within 1 month, she began spiking high fevers, developed diffuse joint pain, and had marked elevations of ferritin, C-reactive protein, and erythrocyte sedimentation rate, characteristic of AOSD. The cutaneous eruption became more widespread, involving the trunk, scalp, and remainder of the extremities, with diffuse thickening of the skin with papular and linear hyperpigmentation and accentuation. Biopsies from several locations showed focal hyperkeratosis associated with dyskeratotic keratinocytes with a peculiar, distinctive distribution in the upper epidermis and cornified layers. In addition, increased dermal mucin was present, with minimal fibroblast proliferation and inflammation. This unusual combination of diffuse dermal mucinosis and a unique pattern of dyskeratosis can present a challenge in generating an accurate differential diagnosis, and may represent an unusual response to chronic scratching or be a distinctive histologic manifestation of AOSD. PMID- 17414448 TI - Cutaneous balloon cell dermatofibroma (fibrous histiocytoma). AB - Dermatofibroma (DF) or cutaneous fibrous histiocytoma is a common benign skin tumor that exhibits multiple, distinct histologic variants. Although clear cell DF has been described in the literature, balloon cell degeneration causing a clear cell DF phenotype has been not been reported to date. Herein, we describe the clinicopathologic findings of balloon cell DF arising on the heel of a 43 year-old man. Clinically, it presented as enlarging tan-white, ulcerated, firm 1.5 cm nodule, clinically suspected to be pyogenic granuloma. Excisional biopsy revealed a circumscribed fibrous tumor populated by mostly clear and spindle cells. A zonal arrangement separated the varied tumor cells where the most superficial, polypoid area showed large, clear polygonal balloon cells; the mid dermal zone demonstrated a transition between balloon cells, epithelioid cells, and spindle cells; and the deep dermal zone had storiform arrangement of spindle cells, with the fascicles separated by coarse collagen bundles. A CD10+ > CD68+ > Factor XIIIa+ immunophenotype was identified with negative immunolabeling for S 100 protein, HMB-45, cytokeratin AE1/AE3, desmin, smooth muscle actin, lysozyme, and leukocyte common antigen (LCA). Ultrastructurally, the clear tumor cells were filled with multiple, empty, nonmembrane bound vacuoles of varying size. No recurrence has been described after complete excision and 7 months of follow up. DF with balloon cell change, likely secondary to persistent irritation, should be added to the differential diagnosis of cutaneous primary and metastatic neoplasms showing balloon cell degeneration such as balloon cell melanocytic nevi and renal cell carcinoma, respectively. PMID- 17414450 TI - Clear-cell basal cell carcinoma: differentiation from other clear-cell tumors. PMID- 17414451 TI - Challenge. Vestibular papilloma of the vulva. PMID- 17414452 TI - Contrary view: the breast is not an organ per se, but a distinctive region of skin and subcutaneous tissue. PMID- 17414455 TI - Reexcision perineural invasion. PMID- 17414456 TI - Reexcision perineural invasion. PMID- 17414457 TI - A phase I study of gemcitabine and uracil-ftorfar (UFT)/leucovorin. AB - BACKGROUND: Gemcitabine and uracil-ftorfar (UFT) are nucleotide analogs with overlapping clinical activity and complementary mechanisms of action. The primary objective of this study was to determine the maximum tolerated dose of UFT/leucovorin and gemcitabine. METHODS: The major eligibility criteria included a diagnosis of nonhematologic cancer with no conventional effective therapy, normal organ function, and Eastern Cooperative Oncology Group performance status of 0-2. The starting doses were 600 mg/m2 gemcitabine weekly for 3 of 4 weeks and 150 mg/m2 UFT daily and 30 mg leucovorin 3 times a day, both for 21 days. Cycles were repeated every 28 days. RESULTS: Twenty-eight patients (male:female 13:15) were treated. The median number of cycles per patient was 3 (range, 0-17). Two of 3 patients on dose level 4 (250 mg/m2 UFT, 800 mg/m2 gemcitabine) developed dose limiting toxicities consisting of hand-foot syndrome and infection. CONCLUSIONS: The recommended doses for phase II trials are 800 mg/m2 gemcitabine days 1, 8, and 15 and 200 mg/m2 UFT per day and 90 mg leucovorin per day on days 1 through 21. PMID- 17414458 TI - QTc monitoring during a phase I study: experience with SR271425. AB - OBJECTIVE: SR271425 is a thioxanthone cytotoxic drug that induces dose-related cardiac electrophysiologic changes in preclinical models. A phase I trial was conducted to determine the maximally tolerated dose and safety profile, notably cardiac events. METHODS: SR271425 was administered weekly as a 2-hour single intravenous infusion with a fixed 30 mg/m2 increment at each dose level (DL). A sustained cardiac evaluation was performed. ECG parameters were evaluated at bedside by an investigator or a cardiologist, as well as by central reading for dose limiting toxicity (DLT) determination. RESULTS: Sixteen patients were treated. Five DLs were explored, from 75 mg/m2/wk to 195 mg/m2/wk. Fourteen patients (87.5%) experienced noncardiac adverse events related to treatment; only 2 patients presented grade 3 toxicity (nausea/vomiting and GGT increase) and no grade 4 toxicities were reported. Asymptomatic grade 1 or 2 QTcF prolongations were observed in 5 patients during central readings, and in 4 cases at bedside. One QTc-DLT, registered at bedside (grade 2), was unconfirmed at central reading, while another QTc-DLT, not noted at bedside, was highlighted by central reading. No arrhythmias or QRS prolongations were observed. CONCLUSIONS: The maximum tolerated dose of SR271425 was not reached in this trial due to early termination of the trial, not related to cardiac toxicity, following the termination of the development program by the sponsor. Sustained ECG monitoring is quite feasible in oncology phase I trials, but discrepancies between bedside and central evaluation could lead to conflicting decisions for management of patient care. PMID- 17414459 TI - Phase III study of standard combination versus rotating regimen of induction chemotherapy in patients with hormone insensitive metastatic breast cancer: an Eastern Cooperative Oncology Group Intergroup Study (E3185). AB - OBJECTIVE: The objective of this multicenter phase III trial was to study the impact on time to treatment failure (TTF) and survival of cyclophosphamide, Adriamycin, and 5-fluorouracil (CAF) versus CAF/thiotepa, Adriamycin, vinblastine, and Halotestin (TsAVbH), a partially noncross-resistant regimen used in a rotating schedule in the treatment of hormone insensitive metastatic breast cancer in accordance with the Goldie and Coldman hypothesis. METHODS: Three hundred forty-three patients received 6 cycles of induction treatment with one of 2 regimens. Patients with estrogen receptor-negative tumors or those with estrogen receptor-positive or estrogen receptor-unknown tumors with demonstrated unresponsiveness to hormone treatment were eligible. Complete responders were randomized to either observation or maintenance therapy with cyclophosphamide, methotrexate, 5-fluorouracil, prednisone, tamoxifen, and Halotestin (CMF[P]TH). Patients with partial response or stable disease on completion of induction therapy were maintained on CMF plus Halotestin. RESULTS: There were no differences in the primary end point of TTF (median 7.3 and 7.4 months, respectively). There was a significant difference in TTF and survival by duration of disease-free interval: a median of 8.8 and 21.2 months for those with a disease-free interval of > or =2 years versus 6 to 8 and 13.3 months for those with a disease-free interval <2 years (P = 0.016 and <0.001), respectively. Toxicity of the 2 treatment regimens was similar. CONCLUSION: There were no differences observed in TTF, survival, and toxicities between the 2 treatment arms, both of which contained doxorubicin (Adriamycin) as the most active agent. The results of observation versus maintenance in complete responders were reported separately. PMID- 17414460 TI - The rates of chemotherapy-induced amenorrhea in patients treated with adjuvant doxorubicin and cyclophosphamide followed by a taxane. AB - OBJECTIVE: Adjuvant chemotherapy in premenopausal women with breast cancer may induce amenorrhea, which can affect fertility, choice of hormonal therapy, and increase the risk of late toxicity. The incidence of chemotherapy-induced amenorrhea (CIA) resulting from doxorubicin and cyclophosphamide (AC) followed by a taxane (T) is poorly characterized. METHODS: We retrospectively surveyed women who were premenopausal and less than age 50 at initiation of chemotherapy to determine the rates of CIA in women receiving AC followed by T compared with AC alone. RESULTS: One hundred ninety-one eligible women completed the survey. The rate of CIA in women who received AC followed by T was 64% (95% confidence interval [CI] = 55-72%) compared with 55% (95% CI = 43-66%) in AC alone. As expected, CIA rates were higher in older than younger women (82% vs. 55%, P = 0.004). Multivariate logistic regression analysis revealed that age >40 was associated with a 4.6-fold increased risk of CIA (95% CI = 1.7-12.1, P = 0.002). It also revealed that receiving T after AC was associated with an odds ratio of 1.9 for CIA as compared with receiving AC alone (95% CI = 1.0-3.5, P = 0.05). Despite > or =6 months of amenorrhea, many women < or =40 resumed menses (40%). CIA was more likely to be irreversible in those >40. The addition of taxanes did not alter the rate of reversibility for the group as a whole (P = 0.36). CONCLUSIONS: Older age and the addition of taxane to AC increased the risk of CIA and the amenorrhea was more likely to be irreversible for women >40. Women < or =40 often resume menstruation even after 6 months of amenorrhea, and the addition of T does not play a role. Subsequent resumption of menstrual function must be considered when initiating appropriate hormonal therapy. PMID- 17414461 TI - A phase II trial of vinorelbine and pegylated liposomal doxorubicin in patients with pretreated metastatic breast cancer. AB - The objective of the present study is to investigate the clinical efficacy and tolerance of vinorelbine and pegylated liposomal doxorubicin (PLD) in patients with metastatic breast cancer (MBC) with prior taxane and/or anthracycline treatment. A total of 25 patients who previously received taxane- and/or anthracycline-based chemotherapy as first- and/or second-line treatment of MBC were entered into the study and were treated with 20 mg/m2 vinorelbine on day 1 and 8 and 30 mg/m2 PLD on day 1 every 3 weeks. All were evaluated for efficacy, quality of life, and tolerance. Three complete and 6 partial responses were observed in 25 patients for an objective response rate of 36% (95% confidence interval: 17-55%). Eight patients (32%) had stable disease of not less than 3 months and 8 patients (32%) had disease progression. The median progression-free survival was 6.7 months (range, 2-18 months), and the median overall survival was 13.2 months (range, 3-31 months). Severe toxicities (grade 3 or above) were neutropenia (16%) and mucositis (8%). The health-related quality of life assessed before each cycle by specific questionnaire did not differ significantly over the treatment period. The combination of vinorelbine and PLD for anthracycline- and/or taxane-pretreated patients with MBC is an active and safe regimen that does not compromise the quality of life. PMID- 17414462 TI - Activity and tolerability of a combined palliative chemotherapy with mitomycin C, folinate, and 5-Fluorouracil in patients with advanced breast cancer after intensive pretreatment: a retrospective analysis. AB - OBJECTIVES: The aim of this retrospective study was to evaluate the activity and toxicity of a combined chemotherapy containing mitomycin, folinate, and 5 fluorouracil (MiFoFU) in patients with advanced metastatic breast cancer and reduced performance status, ie, elderly patients or heavily pretreated patients. METHODS: We studied the charts of 76 patients with progressive metastatic breast cancer who received MiFoFU chemotherapy at our institution between 1997 and 2003. Primary end points were response and time-to-progression (TTP); secondary end points were overall survival (OAS) and tolerability. RESULTS: Median age was 57 years. Seventeen patients had > or =2 palliative cytostatic treatments before; 19 patients were older 65 years. Patients received a median of 6 cycles. Clinical benefit rate was 58%. After MiFoFU, median TTP and OAS were 8 months and 14 months, respectively. Main nonhematologic toxicity was stomatitis (grade I/II, 21%) and diarrhea (grade I/II, 37%). Grade III/IV hematotoxicity was seen in 18 patients (24%). CONCLUSIONS: A combined MiFoFU chemotherapy is a well-tolerated treatment option in the palliative therapy for patients with metastatic breast cancer. In particular, the favorable efficacy/toxicity ratio in intensively pretreated or elderly patients makes this combination a reasonable alternative within these settings. PMID- 17414463 TI - Recurrence rates and analysis of close or positive margins in patients treated without re-excision before radiation for breast cancer. AB - PURPOSE: This study examines the risk of local recurrence in a group of patients accepted for radiation therapy after breast-conserving surgery despite having a close or positive resection margin. METHODS AND MATERIALS: Two hundred patients with early-stage breast cancer were treated by radiation with a nonnegative margin < or =2 mm from January 1974 to September 2001. The median age was 61 years. Margins were positive in 29% and close (< or =2 mm) in 71%. The median dose was 64 to 66 Gy. The median follow up was 5.9 years. RESULTS: The number of resection margins close or positive was 1 in 73% of patients, 2 in 14%, 3 in 1%, and unknown in 12%. The margin location was 23% anterior, 24% posterior, 12% medial, 12% lateral, 17% superior, and 12% inferior. Reasons for not reexcising were advanced age/comorbidities in 7%, anterior location under skin in 25%, posterior location to muscle in 15%, focal involvement in 13%, no extensive intraductal component in 5%, surgeon refusal in 15%, and patient refusal in 20%. There was a strong association between an anterior or posterior margin location and the rationale of no additional breast tissue at the margin to reexcise before radiation. The risk of local recurrence at 5 and 10 years was 3% and 5%, respectively. CONCLUSIONS: Further research of close and positive margins is needed to validate features identified in this series, particularly nonbreast tissue anatomic margins, that are associated with low risks of local recurrence after radiation. PMID- 17414464 TI - Placement technique and the early complications of balloon breast brachytherapy: Magee-Womens Hospital experience. AB - BACKGROUNDS AND OBJECTIVES: Open (OT) and percutaneous closed (PCT) techniques have been described for placement of the MammoSite catheter to deliver accelerated partial breast brachytherapy. We report early complications of both techniques. METHODS: A total of 125 patients underwent catheter placement for MammoSite high-dose rate brachytherapy, with 108 patients successfully completing treatment. The OT was used in 85 patients and PCT in 40 patients. The mean distance between the balloon surface and breast skin was 1.44 cm and 1.31 cm, respectively. Average skin dose was 278 cGy in the OT group and 295 cGy in the PCT group (P > 0.05). Average gross specimen size was 43.16 cm3 in the OT group and 62.19 cm3 in the PCT group. Median follow-up was 11 months for the OT group and 5 months for the PCT group. RESULTS: In 17 cases, the catheter was subsequently removed without the patient completing treatment. Two of the patients in the OT group (3%) developed a delayed abscess. The overall incidence of persistent seroma (>6 months) was 20% with all occurring in the OT group, 30% of those patients. There were no acute skin toxicities higher than grade 2. The overall cosmesis is excellent or good in 95% of patients. CONCLUSION: Despite short follow-up and a small sample size in this study, it seems that the MammoSite brachytherapy was well tolerated by patients with early stage breast cancer when using either the OT or PCT. PMID- 17414465 TI - The impact of involved field radiation therapy in the treatment of relapsed or refractory non-Hodgkin lymphoma with high-dose chemotherapy followed by hematopoietic progenitor cell transplant. AB - BACKGROUND: Patients with refractory/relapsed non-Hodgkin lymphoma (NHL) often receive high-dose chemotherapy (HDCT) followed by hematopoietic progenitor cell transplant (HPCT) as salvage therapy. We examined the role of involved field radiation therapy (IFRT) in this setting. METHODS: The records of 167 patients with refractory/relapsed NHL who underwent HDCT followed by HPCT between February 1990 and November 2003 were reviewed. Fifty-three patients received IFRT and 114 did not receive IFRT in the peritransplant period. RESULTS: Eighty patients were alive at the time of analysis with a median follow up for alive patients of 4.5 years in the no IFRT group and 4.2 years in the IFRT group (P = 0.53). Patients undergoing IFRT were more likely to have bulky (P = 0.02) and extranodal (P= 0.04) disease at initial diagnosis. There was no significant difference between the treatment groups regarding mortality in the first 100 days after HPCT (P = 0.31). Five-year overall survival rates were 46.7% for the no IFRT group and 40.0% for the IFRT group (P= 0.15). Disease-free survival was significantly worse for patients receiving IFRT (P = 0.02); however, when considering local control, the addition of IFRT resulted in a 5-year rate similar to that for patients who did not receive IFRT (68.6% vs. 72.0% respectively, P= 0.73). CONCLUSIONS: Although disease-free survival was inferior in patients who received IFRT, despite more adverse clinical features the use of IFRT resulted in similar rates of local control and overall survival compared with those who did not receive IFRT. The use of IFRT was not associated with an increase in the risk of acute mortality or late events. PMID- 17414466 TI - Variations in chemotherapy and radiation therapy in a large nationwide and community-based cohort of elderly patients with non-Hodgkin lymphoma. AB - OBJECTIVE: The objective of this study was to determine the variations in the use of chemotherapy and radiation therapy in a large nationwide and population-based cohort of older patients with non-Hodgkin lymphoma (NHL). METHODS: We studied a retrospective cohort of 13,570 patients diagnosed with incident NHL at age > or =65 in 1992 through 1999 identified from the Surveillance, Epidemiology and End Results-Medicare-linked database. RESULTS: Of 13,570 patients with NHL, 52% received chemotherapy within 6 months of diagnosis. A higher proportion (61%) of patients aged 65 to 69 received chemotherapy than those > or =80 (39%). Forty three percent of black patients received chemotherapy compared with whites (52%) and other ethnicities (54%). A greater proportion of patients with lower comorbidity scores received chemotherapy than those with higher comorbidity scores. In multivariate analysis, there was no significant association between the receipt of chemotherapy and gender or year of diagnosis, but blacks had statistically significant lower odds of receiving chemotherapy than white patients (odds ratio = 0.66, 95% confidence interval: 0.55-0.80). Significant predictors of receiving chemotherapy were age, ethnicity, comorbidity, tumor stage, nodal site, geographic area, and radiation therapy. The percentages of patients receiving radiation therapy, chemotherapy, or both were 11%, 39%, and 13%, respectively; 37% of patients with NHL did not receive any of these treatments within 6 months of diagnosis. CONCLUSION: There were significant variations in the use of chemotherapy by patient and tumor characteristics and geographic areas. A substantial number of patients with NHL did not receive either chemotherapy or radiation therapy, suggesting that the rate for receipt of these therapies may be improved. PMID- 17414467 TI - A phase II trial of accelerated multimodality therapy for locoregionally advanced cancer of the esophagus and gastroesophageal junction: the impact of clinical heterogeneity. AB - OBJECTIVES: This is a report of mature results from a phase II trial of an accelerated multimodality treatment program for locoregionally advanced cancer of the esophagus and gastroesophageal junction with a focus on the impact of clinical heterogeneity on outcomes. A split course of pre- and postoperative hyperfractionated radiation therapy and concurrent chemotherapy was used in an effort to limit perioperative mortality. METHODS: Eligibility required a diagnosis of esophageal or gastroesophageal junction cancer and an esophageal ultrasound stage of at least T3, N1, or M1A. Patients received a 12-day induction course of radiation (1.5 Gy twice a dose to a dose of 30 Gy) concurrent with 4 day continuous intravenous infusions of cisplatin (20 mg/m2 per day) and 5 fluorouracil (1000 mg/m2 per day) beginning on day 1. Surgery followed in 4 to 6 weeks followed 6 to 10 weeks later by a second, identical course of chemoradiotherapy. RESULTS: From October 1999 through March 2003, 93 patients were enrolled; 96% were white, 86% male, and 83% had adenocarcinoma. Resection was possible in 83 patients (89%) with 4 (5%) perioperative deaths. With a median follow up of 50 months (range, 34-72 months), the 3-year projected overall survival rate is 27.9%, freedom from recurrence 30.5%, and distant metastatic control 32.4%. Locoregional control in resected patients is 86%. Freedom from recurrence and distant control were significantly better in patients with 1) earlier pretreatment clinical stage, 2) earlier postinduction pathologic stage, 3) squamous cell cancer, and 4) a pathologic response. CONCLUSIONS: This accelerated multimodality treatment program is feasible and perioperative mortality proved acceptable. Despite excellent locoregional control, freedom from recurrence, and overall survival proved disappointing reflecting the frequency of distant metastases. Heterogeneity in patient populations makes comparisons with similar nonrandomized experiences problematic. PMID- 17414468 TI - T3 and T4 true vocal cord squamous carcinomas treated with external beam irradiation: a single institution's 35-year experience. AB - PURPOSE: The purpose of this study was to report the outcomes after external-beam radiotherapy (RT) for selected T3 and T4 vocal cord squamous cell carcinomas. METHODS AND MATERIALS: One hundred nine patients with previously untreated T3 and T4 squamous cell carcinomas of the glottic larynx were treated with curative intent in this Institutional Review Board-approved outcome study using continuous course RT alone (106 patients) or followed by a planned neck dissection (3 patients) between September 1966 and June 2002. Patients selected for such treatment had relatively low-volume, unilateral disease. Patients were staged according to the recommendations of the American Joint Committee on Cancer (AJCC) as follows: T3N0, 68 patients (62%); T3N1, 14 patients (13%); T3N2B, 5 patients (5%); T4N0, 17 patients (16%); T4N1, 4 patients (4%); and T4N2B, 1 patient. RESULTS: The 5-year outcomes after treatment were: local control for stage T3 and T4, 78% and 81%; locoregional control for AJCC stage III and IVa, 62% and 78%; distant metastasis-free survival for AJCC stage III and IVa, 97% and 100%; cause specific survival for AJCC stage III and IVa, 84% and 87%; and overall survival for AJCC stage III and IVa, 52% and 67%, respectively. Severe complications occurred in 13 patients (12%). CONCLUSIONS: The results of this retrospective study compare favorably with those published elsewhere in the literature for T3 and T4 vocal cord carcinomas. Local control and ultimate cure probabilities will hopefully improve further with the addition of concomitant chemotherapy to RT for larger tumors. PMID- 17414469 TI - Prognostic significance of paraglottic space invasion in T2N0 glottic carcinoma. AB - PURPOSE: The purpose of this study was to evaluate the prognostic significance of paraglottic space invasion determined with pretreatment computed tomography (CT) for patients with T2N0 glottic carcinoma treated with radiotherapy (RT). METHODS: Eighty patients with T2N0 glottic squamous cell carcinomas treated with definitive RT between 1983 and 2002 who had pretreatment CT information available regarding paraglottic space invasion were analyzed. Follow up ranged from 0.14 to 18 years (median, 7.1 years). No patient was lost to follow up. RESULTS: Five year outcomes were: local control, 78%; local control with larynx preservation, 80%; relapse-free survival, 71%; and cause-specific survival, 91%. Multivariate analyses of various parameters (including vocal cord mobility, paraglottic space invasion, supraglottic invasion, and subglottic extension) showed that only subglottic extension significantly influenced the probability of cure. CONCLUSION: Paraglottic space invasion is likely associated with increased tumor volume and, by itself, is not necessarily associated with poorer outcome after definitive RT. PMID- 17414470 TI - The effect of dental artifacts, contrast media, and experience on interobserver contouring variations in head and neck anatomy. AB - OBJECTIVES: To investigate interobserver variability in the delineation of head and-neck (H&N) anatomic structures on CT images, including the effects of image artifacts and observer experience. METHODS: Nine observers (7 radiation oncologists, 1 surgeon, and 1 physician assistant) with varying levels of H&N delineation experience independently contoured H&N gross tumor volumes and critical structures on radiation therapy treatment planning CT images alongside reference diagnostic CT images for 4 patients with oropharynx cancer. Image artifacts from dental fillings partially obstructed 3 images. Differences in the structure volumes, center-of-volume positions, and boundary positions (1 SD) were measured. In-house software created three-dimensional overlap distributions, including all observers. The effects of dental artifacts and observer experience on contouring precision were investigated, and the need for contrast media was assessed. RESULTS: In the absence of artifacts, all 9 participants achieved reasonable precision (1 SD < or =3 mm all boundaries). The structures obscured by dental image artifacts had larger variations when measured by the 3 metrics (1 SD = 8 mm cranial/caudal boundary). Experience improved the interobserver consistency of contouring for structures obscured by artifacts (1 SD = 2 mm cranial/caudal boundary). CONCLUSIONS: Interobserver contouring variability for anatomic H&N structures, specifically oropharyngeal gross tumor volumes and parotid glands, was acceptable in the absence of artifacts. Dental artifacts increased the contouring variability, but experienced participants achieved reasonable precision even with artifacts present. With a staging contrast CT image as a reference, delineation on a noncontrast treatment planning CT image can achieve acceptable precision. PMID- 17414471 TI - A simple model predicts freedom from biochemical recurrence after low-dose rate prostate brachytherapy alone. AB - OBJECTIVE: The objective of this study was to describe a simple model that predicts freedom from biochemical recurrence (FFBR) in men with prostate cancer after treatment with low-dose rate prostate brachytherapy (LDRPB) alone. MATERIALS AND METHODS: One hundred thirty-two men were treated with LDRPB alone between September 1997 and April 2001. Sixty-four percent of men had low-risk disease (prostate-specific antigen [PSA] <10, Gleason <7, and T stage or =10, Gleason > or =7, or T stage T2b). The dosimetric quantifier D90 was calculated from a computed tomography scan performed 1 month after LDRPB. The percent positive biopsies (PPB) were determined for all patients. FFBR was estimated using the product limit method. All P values are 2-sided. RESULTS: The median follow-up is 65 months. The median D90 is 138 Gy (range, 47-221 Gy). Fourteen men have developed evidence of biochemical relapse at a median of 27 months (range, 6-42 months). The 5-year FFBR rate for the entire cohort is 88%. On univariate analysis, variables found to be associated with FFBR included: PSA, Gleason score, T stage, risk group, PPB, and D90. Multivariate analysis indicated that D90, PPB, and risk group were independently associated with FFBR. Patients were categorized based on the following 3 adverse prognostic factors: D90 <140 Gy, PPB > or =50%, and intermediate-risk group. Group 1 (0 factors, n = 30), group 2 (1 factor, n = 72), and group 3 (> or =2 factors, n = 30) patients had 5-year FFBR rates of 100% (+/ 0%), 92% (+/-6%), and 67% (+/-18%) (P < 0.0001). CONCLUSIONS: We have developed a simple, robust model based on implant quality and disease factors that predicts FFBR in men with prostate cancer treated with LDRPB alone. PMID- 17414472 TI - Long-term outcome of stage I seminoma. AB - PURPOSE: To report on long-term outcomes among patients with stage I seminoma treated by orchiectomy with or without adjuvant radiation. MATERIALS AND METHODS: A retrospective review of medical records of patients treated between 1974 and 2002 was undertaken to identify factors associated with patient outcomes. RESULTS: With a median follow-up of 7.7 years, 80% (4 of 5) of the surveillance group experienced a disease relapse, while only 3% (2 of 70) in the radiation therapy group had disease relapse. This difference in relapse rates was statistically significant, but there was no significant difference in overall survival between the 2 groups. There was a significant relationship between patient age and disease relapse, whereby all of the relapses were seen in patients younger than 36 years at diagnosis (P = 0.03). Of the total 75 patients, 7 (9%) developed second primary tumors. Six of them (6 of 7) were treated with adjuvant radiation, and 1 patient (1 of 7) was on surveillance. CONCLUSION: In this study, risk of relapse was significantly associated with surveillance and in patients younger than 36 years at diagnosis. These results suggest that surveillance can only be safely adopted for patients who can be followed up closely. We consider adjuvant radiation a very effective choice despite the low risk of associated secondary malignancies. PMID- 17414473 TI - Acute intracerebral hemorrhage in intravascular lymphoma: a serious infusion related adverse event of rituximab. PMID- 17414474 TI - Recurrent hypoglycemia associated with poorly differentiated carcinoma of the liver. PMID- 17414475 TI - Sport and exercise medicine: a timely specialty development. PMID- 17414476 TI - Inter-association task force recommendations on emergency preparedness and management of sudden cardiac arrest in high school and college athletic programs: a consensus statement. AB - OBJECTIVE: To assist high school and college athletic programs prepare for and respond to sudden cardiac arrest (SCA). This consensus statement summarizes our current understanding of SCA in young athletes, defines the necessary elements for emergency preparedness, and establishes uniform treatment protocols for the management of SCA. BACKGROUND: SCA is the leading cause of death in young athletes. The increasing presence of and timely access to automated external defibrillators (AEDs) at sporting events provides a means of early defibrillation and the potential for effective secondary prevention of sudden cardiac death. An Inter-Association Task Force was sponsored by the National Athletic Trainers' Association to develop consensus recommendations on emergency preparedness and management of SCA in athletes. RECOMMENDATIONS: Comprehensive emergency planning is needed for high school and college athletic programs to ensure an efficient and structured response to SCA. Essential elements of an emergency action plan include establishing an effective communication system, training of anticipated responders in cardiopulmonary resuscitation and AED use, access to an AED for early defibrillation, acquisition of necessary emergency equipment, coordination and integration of onsite responder and AED programs with the local emergency medical services system, and practice and review of the response plan. Prompt recognition of SCA, early activation of the emergency medical services system, the presence of a trained rescuer to initiate cardiopulmonary resuscitation, and access to early defibrillation are critical in the management of SCA. In any collapsed and unresponsive athlete, SCA should be suspected and an AED applied as soon as possible for rhythm analysis and defibrillation if indicated. PMID- 17414477 TI - Physical activity after total joint replacement: a cross-sectional survey. AB - OBJECTIVE: To determine the level of physical activity participants are able to perform at a minimum of 1 year after primary total hip or knee replacement. DESIGN: Cross-sectional survey. SETTING: A tertiary care arthroplasty center. PARTICIPANTS: A total of 170 primary total hip and 184 primary total knee arthroplasty patients. INTERVENTIONS: The University of California Los Angeles (UCLA) activity score was mailed to 242 primary hip and 225 primary knee arthroplasty patients. Patients were abstracted from a prospectively tabulated arthroplasty database and pre-selected for good/excellent clinical outcomes as determined by 1 year postoperative Knee Society (KSS) and Harris Hip (HHS) scores. Clinical outcomes including the Oxford Hip/Knee score were collected preoperatively, and at 6 and 12 months postoperatively. Correlations between UCLA scores, demographics, and clinical outcomes were calculated using Pearson's correlation. MAIN OUTCOME MEASUREMENTS: Harris Hip Score, Oxford Hip Score, Knee Society Score, Oxford Knee Score, and UCLA Activity Scale. RESULTS: : Postal survey response rates for hips were 70.2% (170 of 242) and 81.8% for knees (184 of 225). Mean results at postoperative year 1 include: HHS (94.8), Oxford Hip Score (16.6), KSS clinical score (95.9), KSS function score (95.0), and Oxford Knee score (18.2). For both primary total hip arthroplasty and total knee arthroplasty patients, median UCLA score was 6, indicating moderate activity levels at a mean follow-up of 40.7 months for hips and 36.6 months for knees. CONCLUSIONS: UCLA scores indicate the average total joint replacement patient maintains a moderate activity level, and many perform active/very active levels of activity. PMID- 17414478 TI - Generalizability of the sensory organization test in college-aged males: obtaining a reliable performance measure. AB - OBJECTIVE: This study used generalizability theory to (1) determine the reliability of the six Sensory Organization Test (SOT) conditions, and (2) to determine the most effective testing schedule to obtain reliable posture measurements. DESIGN: Generalizability theory was used to determine the most effecting testing protocol to obtain reliable assessments of postural sway in various sensory environments. PARTICIPANTS: Twenty-five recreationally active college-aged males. SETTING: University research laboratory. INTERVENTIONS: None. MAIN OUTCOME MEASUREMENTS: Postural stability as measured by equilibrium score (a percentage measure indicating amount of sway, with lower scores indicating a greater amount of postural sway). Scores were obtained for each of the six sensory conditions across multiple trials, tests, and days. RESULTS: This study showed that performing the SOT twice on a single testing day was sufficient to achieve moderate to good reliability across the six conditions of the SOT. Extending SOT testing to twice on 2 days yielded good to excellent reliability estimates across all SOT conditions. CONCLUSIONS: Taken together, these results indicate that for the purpose of monitoring changes in the postural control system, an individual needs to experience the full complement of SOT testing conditions at least twice. However, if highly reliable baseline measures of SOT performance are necessary, multiple tests performed on multiple days are warranted. PMID- 17414479 TI - Maintenance of plasma volume and serum sodium concentration despite body weight loss in ironman triathletes. AB - OBJECTIVE: To examine the relationship between body weight, plasma volume, and serum sodium concentration ([Na]) during prolonged endurance exercise. DESIGN: Observational field study. SETTINGS: 2000 South African Ironman Triathlon. PARTICIPANTS: 181 male triathletes competing in an Ironman triathlon. MAIN OUTCOME MEASURES: Body weight, plasma volume, and serum ([Na]) change from pre- to postrace. RESULTS: Significant body weight loss occurred (-4.9 +/- 1.7%; P < 0.0001), while both plasma volume (1.0 +/- 11.2%; P = 0.4: NS) and serum [Na] (0.6 +/- 2.4%; P < 0.001) increased from pre- to postrace. Blood volume (-0.6 +/- 6.6%) and red cell volume (-2.6 +/- 5.5%; P < 0.001) decreased in conjunction with the body weight loss. There was a strong correlation between blood and plasma volume change, both as a percentage, and absolute change in fluid volume (r = 0.9; P < 0.001). Body weight change was positively correlated with plasma volume change (r = -0.4; P < 0.001), but inversely correlated with serum [Na] change (r = -0.4; P < 0.001). Plasma volume change was not significantly correlated with serum [Na] change (r = 0.0; NS). Serum [Na] change was inversely correlated with both percentage of red cell volume change (r = -0.2; P < 0.05) and percentage body weight change (r = -0.4; P < 0.001). CONCLUSION: Plasma volume and serum [Na] were maintained in male Ironman triathletes, despite significant (5%) body weight loss during the course of the race. Body weight was not an accurate "absolute" surrogate of fluid balance homeostasis during prolonged endurance exercise. Clinicians should be warned against viewing these three regulatory parameters as interchangeable during an Ironman triathlon. PMID- 17414480 TI - The bone response to non-weight-bearing exercise is sport-, site-, and sex specific. AB - OBJECTIVE: To examine whether skeletal adaptations to chronic non-weight-bearing exercise depend on the type of aquatic exercise (swimming or water polo) as well as on sex (men or women). DESIGN, SETTING, AND PARTICIPANTS: This was a cross sectional study at the Laboratory of Nutrition and Clinical Dietetics, Department of Nutrition and Dietetics, Harokopio University, Athens, Greece. A total of 43 water polo players, 26 swimmers, and 30 sedentary individuals, aged 17 to 34 years, were recruited (52 men, 47 women). MAIN OUTCOME MEASURES: Bone mineral content (BMC) and areal bone mineral density (aBMD) of the total body and of various subregions. RESULTS: : Compared with controls, swimmers had lower leg and total aBMD (P < 0.05), whereas water polo players had lower leg but higher arm and trunk aBMD (all P < 0.05). Swimmers and water polo athletes differed at the arms (men only), trunk, and total body (all higher in water polo players, at P < 0.05). Bone adaptations to water polo playing were unaffected by sex. Female swimmers, but not male swimmers, had 13% higher arm BMC than controls (P < 0.05), whereas male swimmers, but not female swimmers, had 12% lower leg BMC than controls (P < 0.05). CONCLUSIONS: Athletes participating in long-term water polo playing and swimming have substantially different total and regional aBMD. The effect is not mediated by sex in water polo players; however, sex may mediate the differences between swimmers and controls. Whether the observed differences between athlete groups and sexes arise from different bone adaptations to activity or from other factors cannot be answered by the current data. CLINICAL RELEVANCE: Water polo playing may be preferable over swimming for maintaining bone health; both types of aquatic exercise at the elite level of participation, however, have unfavorable effects on the lower limb bones. PMID- 17414481 TI - Increased deposition of sulfated glycosaminoglycans in human patellar tendinopathy. AB - OBJECTIVE: To investigate if the increased proteoglycans in patellar tendinopathy involves a qualitative change in the types of proteoglycans. DESIGN: This is an observational study based on the biochemical analysis of proteoglycans. SETTING: : University Teaching Hospital. PATIENTS: Patellar tendon samples from 12 patients with patellar tendinopathy and 12 healthy controls were collected and proteoglycans were extracted for biochemical analyses. All patients fulfilled the diagnostic criteria of having patellar tendinopathy with well-defined clinical features, more than 6 months of insufficient nonoperative treatment including physiotherapeutic modalities, and verification by ultrasound or magnetic resonance imaging. Twelve control subjects, 10 men and 2 women with an average age of 31 years (range 16 to 38 years), represented patients with anterior cruciate ligament deficiency who were operated on using the healthy patellar tendon as an autograft. The control subjects had no previous history or clinical signs of patellar tendon injury. INTERVENTIONS: The independent variable is the presence of pathological conditions of patellar tendinopathy. MAIN OUTCOME MEASUREMENTS: The dependent variables include the electromobility of proteoglycans, staining intensity of proteoglycan core proteins, and the tissue content of glycosaminoglycan disaccharides. RESULTS: The results indicated that the increased proteoglycans in pathological tissues also exhibited qualitative changes as compared to those in healthy patellar tendons. Dermatan monosulfates were significantly increased in the proteoglycans extracted from the pathological tissues of patellar tendinopathy. CONCLUSIONS: Our results indicate that proteoglycans deposited in the pathological tissues of patellar tendinopathy were oversulfated as compared to healthy tendons, which may represent a new pathological attribute for the understanding of chronic pain in patellar tendinopathy. PMID- 17414482 TI - Hemoglobin levels and athletic performance in elite speed skaters during the olympic season 2006. AB - OBJECTIVE: To test the hypotheses that the hemoglobin (Hb) distribution curve in elite male and female long track speed skaters is not normally distributed and that there is a positive relationship between competitive success and Hb concentration. DESIGN: A venous blood sample was taken before the events from all skaters. The Hb concentration distribution curves of all ranked from 1 to 30 were tested for normality. In addition, individual Hb concentrations were plotted against ranking in the matching events. SETTING: 2006 major championships and Olympic winter games. PARTICIPANTS: All elite male and female speed skaters (217 men and 200 women) competing in major international championships in 2006 and in the Olympic winter games 2006. MAIN OUTCOME MEASUREMENTS: Hb concentration and individual ranking in the matching event. RESULTS: The mean Hb levels in men and women were 15.7 +/- 0.8 g/dL and 14.0 +/- 0.7 g/dL, respectively. The distribution curve in men would meet the criteria for normal distribution when 4 values from 2 skaters with naturally high Hb levels were neglected. In the women, the distribution curve did not meet the criteria for normality because of low frequency in the right side of the distribution curve and a high frequency at the left side. The curve failed to have a steep drop off at the right side. When plotting Hb concentration against ranking, there is no correlation and relationship between Hb concentration and competitive success. CONCLUSIONS: The Hb concentrations are within the normal range for endurance athletes, and there is no indication that the values are titrated toward the upper allowed limit. In addition, there is no relationship between Hb concentration and competitive success in elite speed skaters. PMID- 17414483 TI - Sport-related concussion misunderstandings among youth coaches. AB - OBJECTIVE: To determine the understanding of sport-related concussion among youth sports coaches. DESIGN: Cross-sectional survey. SETTING: The survey was administered at coaches' meetings, following practices, and via mail. PARTICIPANTS: 156 active youth sports coaches, with 5.88 +/- 3.16 years (range 1 22) of coaching experience. INTERVENTIONS: An original survey instrument developed to assess concussion knowledge. Internal validity of the instrument was established prior to the study (Cronbach's alpha = 0.83). MAIN OUTCOME MEASUREMENTS: Scores on the survey instrument reflecting symptom recognition and management knowledge. RESULTS: For the 16 items comprising the symptom recognition survey, the mean number of correct responses was 9.78 +/- 2.07. Previous coaching education was predictive of better symptom recognition. On the true/false portion, between 49.4% and 61.5% of coaches correctly answered the 4 statements. CONCLUSION: This investigation revealed that, among youth sports coaches, coaching education was predictive of the ability to recognize signs and symptoms of sport-related concussion. However, several misconceptions about concussion still exist, highlighting that education regarding concussion is necessary. The presence of qualified health care personnel, such as an athletic trainer, at the youth organization level may enhance early recognition, treatment, and referral of concussions. PMID- 17414484 TI - Athletes' preference for gender of team physician. PMID- 17414485 TI - The effect of wrist guards on wrist and arm injuries among snowboarders: a systematic review. AB - OBJECTIVE: To systematically review studies that examined the effectiveness of wrist guards in preventing wrist injuries among snowboarders. DATA SOURCE: MEDLINE (1966-March 2005), EMBASE (1988-March 2005), Cochrane (2005 Issue 1), Sport Discus (1975-March 2005) were searched using variations of the term "snowboard." PubMED was searched for the year 2005 to capture any recently published studies not yet indexed in MEDLINE. The reference lists of included studies and conference proceedings were also searched. STUDY SELECTION: Studies were included if the number of wrist injuries between wrist guarded and unguarded snowboarders could be ascertained. Randomized controlled trials (RCTs), cohort studies, and case-control studies were included. Six studies were included. DATA EXTRACTION: Information regarding study design, patient characteristics, wrist guard characteristics, data source (for cohort and case-control studies), and results (type and severity of injury, compliance, and adverse events) were extracted. Data were extracted by one reviewer and checked by a second reviewer. DATA SYNTHESIS: Data from RCTs and cohort studies were expressed as relative risks with odds ratios presented for case-control studies. The risk of wrist injury (RR: 0.23; 95% CI: 0.13, 0.41), wrist fracture (RR: 0.29; 95% CI: 0.10, 0.87), and wrist sprain (RR: 0.17; 95% CI: 0.07, 0.41) was significantly reduced with the use of wrist guards. Among the case-control studies, wrist guards significantly lowered the odds of sustaining a wrist injury (OR: 0.46; 95% CI: 0.35, 0.62). In an RCT, the risk ratio suggested wrist guards protect the shoulder (RR: 0.22; 95% CI: 0.01, 4.60). Nonexperimental data suggested the possibility that wrist guards may increase the risk of finger and elbow-shoulder injuries. CONCLUSIONS: Wrist guards reduce the risk of wrist injuries among snowboarders. For every 50 snowboarders who were a wrist guard, one wrist injury will be averted. Future research should focus on determining the optimal type of wrist guard and if they increase the risk of other upper extremity injuries. PMID- 17414486 TI - Acute, isolated lateral compartment syndrome after ankle inversion injury. PMID- 17414487 TI - Anterior cruciate ligament reconstruction in a blind athlete: a case report. PMID- 17414488 TI - Klippel-Feil syndrome in a cheerleader. PMID- 17414489 TI - An unusual presentation of synovial chondromatosis of the knee: a case report. PMID- 17414491 TI - Topical ketoprofen patch for ankle sprain. PMID- 17414490 TI - Urachal cyst: unusual presentation in an adolescent after intense abdominal exercise. PMID- 17414493 TI - Physiological roles of muscle-derived interleukin-6 in response to exercise. AB - PURPOSE OF REVIEW: To discuss recent findings with regard to the regulation of muscle-derived interleukin-6 as well as the possible physiological and metabolic roles of interleukin-6 in response to exercise. RECENT FINDINGS: Contraction induced transcription and release of interleukin-6 is primarily regulated by an altered intramuscular milieu in response to exercise. Accordingly, changes in calcium homeostasis, impaired glucose availability and increased formation of reactive oxygen species are all associated with exercise and capable of activating transcription factors known to regulate interleukin-6 synthesis. Acute interleukin-6 administration to humans increases lipolysis, fat oxidation and insulin-mediated glucose disposal. Adenosine monophosphate-activated protein kinase activation by interleukin-6 appears to play an important role in modulating some of these metabolic effects. Interleukin-6 facilitates an antiinflammatory milieu and may exert some of its biological effects via inhibition of the proinflammatory cytokine tumor necrosis factor-alpha. SUMMARY: The discovery of contracting muscle as a cytokine-producing organ opens a new paradigm: skeletal muscle is an endocrine organ that in response to contractions produces and releases 'myokines', which subsequently can modulate the metabolic and immunological response to exercise in several tissues. In our view, interleukin-6 may be one of several myokines. PMID- 17414494 TI - Metabolic response to injury and role of anabolic hormones. AB - PURPOSE OF REVIEW: To provide a short review of the literature describing the hypermetabolic response to injury and potential treatments. Associated findings include changes in inflammatory mediators and secreted hormones. RECENT FINDINGS: Treatments should be aimed at decreasing the response and potentially the use of anabolic agents. Of note, recent interest in the hyperglycemic response to injury and insulin treatment will be highlighted. SUMMARY: The current metabolic care of the burned patient including nutrition is now being unfolded. It is relatively clear that anabolic treatment should be considered in all those with severe injury. Timing of the treatment, however, is still a topic of discussion. PMID- 17414495 TI - Emerging results of anticatabolic therapy with ghrelin. AB - PURPOSE OF REVIEW: This review summarizes recent developments in research into anticatabolic therapies with ghrelin. Potential conditions in which ghrelin treatment may be useful include cachexia, anorexia and ageing. We highlight a number of intriguing basic topics related to the anticatabolic effects of ghrelin. RECENT FINDING: Repeated administration of ghrelin to patients with congestive heart failure or chronic obstructive pulmonary disease improved appetite, body composition, muscle wasting and functional capacity in open-label pilot studies. An acute, randomized, placebo-controlled, crossover clinical trial of cancer patients with anorexia revealed marked increases in energy intake following treatment. The effects of ghrelin treatment in patients with anorexia nervosa are controversial. Basic research studies have extended our understanding of the upstream regulation of neuropeptide Y/agouti-related protein signalling and the central control of adipocyte metabolism. In addition, alterations in fat free mass may play a role in ghrelin regulation. SUMMARY: A number of studies are currently evaluating the anticatabolic effects of ghrelin in the treatment of various diseases, including cachexia, anorexia and age-related disorders. These studies will hopefully lead to the development of novel clinical applications for ghrelin treatment. These studies have also facilitated a better understanding of the molecular basis of the anticatabolic effects of ghrelin. PMID- 17414496 TI - Practicalities of nutrition support in the intensive care unit. AB - PURPOSE OF REVIEW: Nutrition support improves clinical outcomes in the critically ill and our understanding of its effects has advanced significantly over the last few years. Three recently published evidence-based guidelines have made generally consistent and thorough recommendations to assist clinicians in providing nutrition support. This review will focus on various aspects of these recommendations, concentrating on the practicalities of nutrition support in the intensive care unit, such as its optimal mode and composition. RECENT FINDINGS: Enteral nutrition is preferred to parenteral nutrition unless there is a major gut condition which will delay commencement of enteral nutrition. Nasogastric feeding should begin within 24 h, but if intolerance develops, small bowel feeding or pro-motility drugs (erythromycin or metoclopramide) should be attempted before resorting to supplementary parenteral nutrition. Enteral nutrition should not routinely be supplemented with arginine or glutamine, but it should contain a package of eicosapentaenoic acid, gamma-linolenic acid and antioxidants if the patient has acute lung injury or sepsis. Parenteral nutrition should be glutamine supplemented and the prescription should be limited in energy to avoid hyperglycemia. Whether using enteral nutrition or parenteral nutrition, most patients should receive intravenous selenium, and may also need zinc and copper supplementation. SUMMARY: Intensive care unit patients should have nutrition support based on recent evidence-based guidelines with a preference for nasogastric feeding. If intolerance occurs, pro-motility drugs and small bowel feeding should be attempted. Clinicians should also consider carefully the composition of the nutrition support regimen with regard to lipid content (especially eicosapentaenoic acid and gamma-linolenic acid), antioxidants, glutamine and other micronutrients. PMID- 17414497 TI - Use of small-bore feeding tubes: successes and failures. AB - PURPOSE OF REVIEW: Early enteral nutrition is the preferred option for feeding patients who cannot meet their nutrient requirements orally. This article reviews complications associated with small-bore feeding tube insertion and potential methods to promote safe gastric or postpyloric placement. We review the available bedside methods to check the position of the feeding tube and identify inadvertent misplacements. RECENT FINDINGS: Airway misplacement rates of small feeding tubes are considerable. Bedside methods (auscultation, pH, aspirate appearance, air bubbling, external length of the tube, etc.) to confirm the position of a newly inserted small-bore feeding tube have limited scientific basis. Radiographic confirmation therefore continues to be the most accurate method to ascertain tube position. Fluoroscopic and endoscopic methods are reliable but costly and are not available in many hospitals. Rigid protocols to place feeding tubes along with new emerging technology such as CO2 colorimetric paper and tubes coupled with signaling devices are promising candidates to substitute for the blind placement method. SUMMARY: The risk of misplacement with blind bedside methods for small-bore feeding tube insertion requires a change in hospital protocols. PMID- 17414498 TI - Role of branched-chain amino acids in liver disease: the evidence for and against. AB - PURPOSE OF REVIEW: There is ample evidence that patients with liver disease have an ongoing energy and protein catabolism. Nutritional management in these patients must receive high priority. The administration of branched-chain amino acids to patients with liver disease has been a controversial subject. This review is an update on the data available from various studies involving branched chain amino acids supplementation in patients with chronic liver disease and associated complications. RECENT FINDINGS: This review summarizes the results of nutritional interventions involving branched-chain amino acids supplementation carried out in different centres around the world. It is interesting to note that no toxic effects of branched-chain amino acids supplementation have been reported in any of these trials. SUMMARY: Administration of branched-chain amino acids stimulates hepatic protein synthesis in patients with chronic liver disease and this could contribute significantly to improving their nutritional status, and result in a better quality of life. The beneficial role of branched-chain amino acids supplementation in patients with chronic hepatic encephalopathy has been clearly documented in some studies but the exact mechanism of action is still not clear. PMID- 17414499 TI - Drug-nutrient interactions in elderly people. AB - PURPOSE OF REVIEW: The presence of multiple diseases, polypharmacy, malnutrition, and impaired metabolism in elderly individuals increases the risks of adverse events related to drug-food interactions. Some considerations for elderly people influenced by drug-food interactions are reviewed. RECENT FINDINGS: When investigating pharmacokinetic and pharmacodynamic modifications in the elderly, other factors have to be considered, such as anorexia, dementia, depression, intolerance, gastrointestinal-tract disorders, social and economic factors, reduced abilities (visual and manual) and difficulties in chewing or swallowing. Specific reference is made herein to the health status of the elderly Brazilian population based on the observations of our research group. In addition, the most common diseases (such as cancer, coronary heart disease, dementia, diabetes mellitus, hypertension and osteoporosis), the drugs usually prescribed to treat them, and the adverse nutritional reactions that occur in older patients are summarized. SUMMARY: In order to develop a correct drug prescription plan and nutritional intervention to avoid any kind of undesirable drug-food interaction effect, it is necessary to adequately diagnose the disease and often re-evaluate the chosen treatment, identify disease stages and the necessary therapies to minimize the number of drugs administered, and select a reasonable nutritional assessment. PMID- 17414500 TI - Stability and compatibility of histamine H2-receptor antagonists in parenteral nutrition mixtures. AB - PURPOSE OF REVIEW: To examine the stability and compatibility of histamine H2 receptor antagonists in parenteral nutrition admixtures in order to provide well tolerated and effective therapy. RECENT FINDINGS: Many stability investigations were carried out before 2005, but no critical appraisal of published data has been undertaken. H2-receptor antagonists are stable in most parenteral nutrition admixtures for 2-48 h, with long-term stability up to 4 weeks claimed for a few combinations. Some earlier data are questionable in the light of more recent analytical technology and pharmaceutical knowledge regarding stability assessment. As stability is dependent upon the source of constituents, we have listed all products tested and their respective manufacturers. A general recommendation is made not to store H2-receptor antagonists in parenteral nutrition for more than 24 h at refrigerated temperature before use. SUMMARY: Administration of H2-receptor antagonists in parenteral nutrition admixtures has several clinical and economical advantages, but, in order for these benefits to be well tolerated and efficacious, the stability and compatibility of the drugs, as well as the parenteral nutrition components after mixing, have to be ascertained. Factors that influence stability are assessed, the need for more controlled study protocols is evaluated and recommendations are made for safe storage and administration. PMID- 17414501 TI - Determinants of quality of life in home parenteral nutrition. AB - PURPOSE OF REVIEW: To highlight the most important and salient articles regarding home parenteral nutrition and quality of life published within the last 3 years. RECENT FINDINGS: In recent years, quality of life research in home parenteral nutrition has highlighted the need for a therapy-specific validated questionnaire. Several papers suggest a greater psychological input is required to better understand and evaluate this patient population. Issues surrounding the use of home parenteral nutrition in malignancy have arisen, prompting discussion on ideal timing and candidacy for home parenteral nutrition. Intestinal transplantation is evolving and improving, making it a possible alternative to home parenteral nutrition. Earlier referral is suggested as late referral can result in poorer outcome. SUMMARY: Home parenteral nutrition is a life-sustaining therapy for individuals with intestinal failure. There is now a relatively large amount of research into the quality of life in this population, but more focused measurements (in the form of validated therapy-specific questionnaires) are required to answer questions relating to cancer and intestinal transplantation. PMID- 17414502 TI - Oxidative stress and antioxidant protection in the perinatal period. AB - PURPOSE OF REVIEW: Oxidative stress is implicated in the pathophysiology of disorders during pregnancy, childbirth and the postnatal period, but open questions exist on the effects of antioxidants, the role of oxidative stress during resuscitation and the antioxidative protection of human milk. RECENT FINDINGS: In a randomized controlled trial, fish oil supplementation to pregnant women did not enhance oxidative stress. Two randomized controlled trials supplementing high dosages of both vitamins C and E did not show any reduction in the incidence of preeclampsia. A systematic review and meta-analysis of studies comparing resuscitation of asphyxiated infants either with 100% oxygen or with room air demonstrated significantly lower neonatal mortality with use of room air. Antioxidative properties of human milk were greater in milk from mothers of term than of preterm infants. SUMMARY: Increased levels of oxidative stress and reduced antioxidative capacities may contribute to the pathogenesis of disorders in the perinatal period. Based on the available evidence, supplementation of pregnant women with high dosages of both vitamin C and E cannot be recommended. Asphyxiated newborn infants should be resuscitated with room air rather than with 100% oxygen. Breastfeeding appears beneficial in providing antioxidative protection. PMID- 17414503 TI - Iron requirements, absorption and metabolism in infancy and childhood. AB - PURPOSE OF REVIEW: Iron deficiency is a significant public health problem in young children due to their high iron requirements, and iron supplements are therefore often recommended. During the time period in focus for this review (2005-2006), there have been additional advances in our understanding of the molecular mechanisms of iron absorption and metabolism. It has also been suggested that iron supplements may have adverse effects in children. RECENT FINDINGS: Recently discovered molecules, for example hepcidin, lactoferrin receptor and heme carrier protein may be important for iron metabolism in children. There are possible metabolic interactions between iron and several other minerals. Many studies show that iron deficiency in young children is associated with impaired neurodevelopment but it is not clear whether this can be prevented by iron supplementation. Oral iron supplements given to young children in malarious regions may lead to increased risk of death or severe infections, especially in those who are iron replete. SUMMARY: More research is needed to identify those children who will benefit from iron supplementation and to better determine iron requirements during early life. Clinical trials should include functional outcomes. Better knowledge about molecular mechanisms and nutrient interactions may lead to new diagnostic tests and preventive strategies. PMID- 17414504 TI - Dietary risk factors for development of childhood obesity. AB - PURPOSE OF REVIEW: Controversial information exists on the contribution of several dietary factors for overweight development in childhood, but there is no doubt that obesity prevalence is increasing. We review the most up-to-date information in order to clarify the evidence-based dietary aspects influencing obesity development in children and adolescents. RECENT FINDINGS: Longitudinal studies are the preferred method for analysing the relationship between dietary factors and obesity development. With the exception of infants, there are no conclusive associations between energy intake or diet composition and later overweight development in children. Among formula or mixed-fed infants, the increase in energy intake has been associated with an increased risk of being overweight during childhood. Breastfeeding seems to be a protective factor for later obesity development. In terms of food intake, longitudinal studies have only found a clear and positive association between obesity development and sugar sweetened beverage consumption; this is not the case with snacking, fast food or food portion sizes. Cross-sectional studies have found correlations between being overweight in childhood and buying lunch at school, eating supper while watching television or without family supervision, consuming less energy at breakfast or more at dinner, and missing breakfast. SUMMARY: Results from longitudinal studies must be taken into account in order to design preventive strategies to counteract the increased prevalence of obesity and its consequences in children. Lack of breastfeeding, high early energy intake and high intake of sugar-sweetened beverages seem to be the main dietary factors contributing to obesity development. PMID- 17414505 TI - Nutrition and performance in children. AB - PURPOSE OF REVIEW: Malnutrition in late infancy and childhood remains a significant public health issue in developing nations as well as for those in transition to an industrialized economy. In addition, in these settings and particularly in developed nations, overweight is becoming a very serious threat to both the immediate and the long-term health of children. In this review, we present recent studies that have examined relationships between childhood undernutrition and three general areas of performance: physical activity, cognition and behavior. RECENT FINDINGS: Malnourished children have been shown to have decreased physical activity and endurance, and poorer cognitive function and school performance. Multiple single micronutrient deficiencies, including vitamin B12, thiamin, niacin, zinc and iron, have been associated with poorer cognitive performance. Behavioral problems, including attention deficits, have also been associated with food insufficiency and malnutrition. SUMMARY: The effects of impaired nutritional status during childhood may have long-standing consequences for the health and performance of children during their adult years. PMID- 17414506 TI - Nutritional management of cystic fibrosis patients. AB - PURPOSE OF REVIEW: We critique recent advances in nutritional care of patients with cystic fibrosis focusing on our understanding of the role of fecal elastase as a pancreatic function test, enzyme replacement therapy and bone health. RECENT FINDINGS: Fecal elastase measured by the monoclonal antibody method is a useful screening test for delineating cystic fibrosis patients with pancreatic insufficiency (needing enzyme replacement therapy) from those with pancreatic sufficiency. Cutoff levels, however, have not been clearly defined, particularly in infancy. Newer enzyme products that attempt to improve the intralumenal intestinal pH with bicarbonate or using a combination of unprotected and enteric coated products fail to completely correct nutrient maldigestion. Compromised bone health in cystic fibrosis patients may be due to multiple factors including calcium, vitamin D, and vitamin K deficiencies and lung inflammation. Current recommendations for treatment of bone health in cystic fibrosis are not evidence based. SUMMARY: Fecal elastase is a useful marker of pancreatic function but limitations for this test exist. The presence of an acidic intestinal milieu and hitherto poorly defined intestinal factors may contribute to the failure of current enzyme preparations to correct nutrient assimilation in cystic fibrosis. The many factors that contribute to bone health must be evaluated when developing treatment strategies. PMID- 17414507 TI - Counseling parents on feeding their children. AB - PURPOSE OF REVIEW: The high-nutrient needs of children for normal growth and development are matched by ready access to low-nutrient high-energy foods. Parents are often confused by new and at times conflicting information. This review discusses three recently published papers that offer specific and important dietary information for school-aged children. RECENT FINDINGS: Lactose intolerance is common in some populations and there are misconceptions about dairy intake. Most lactose-intolerant children can consume some dairy products without symptoms. Fruit-juice intake can predict increased weight gain in children, especially those who are already overweight or at risk for being overweight. Hypertension is a serious disease with onset likely in childhood. This paper discusses the importance of dietary sodium as a contributor to the development of hypertension, and the sodium content of children's diets. SUMMARY: Advice to parents on feeding children should be based on the food pyramid and include information on exercise. Recent publications suggest that children consume dairy products, even if lactose intolerant, restrict juice intake, remove sweetened beverages from their diets and reduce sodium consumption. PMID- 17414508 TI - Soy protein for infant feeding: what do we know? AB - PURPOSE OF REVIEW: This review discusses the safety, nutritional adequacy and recommendations for use of soy protein formulae, based mainly on the most relevant reports published during 2005 and 2006. RECENT FINDINGS: Concerns have recently been raised regarding potential risks with soy protein formulae, in particular regarding their high phytoestrogenic isoflavone content. Recent data are insufficient to draw definitive conclusions on safety, but authorities and paediatric societies from several countries recently advised health professionals to use soy protein formulae only in certain cases. Indications for use of soy protein formulae, mainly for prevention and management of food allergy, have also been better defined. SUMMARY: Soy protein formulae ensure normal growth and development in healthy term infants but they have no nutritional advantages over cow's milk protein formulae. Main indications include severe lactose intolerance, galactosaemia and need to avoid foods of animal origin. Soy protein formulae have no role in preventing allergy or in management of nonspecific gastrointestinal symptoms (e.g. infantile colic and regurgitation). They should not be used in preterm infants or infants with food allergy before age 6 months. After 6 months, soy protein formulae may be considered if tolerance to soy protein is established. PMID- 17414509 TI - Bibliography. Current world literature. Anabolic and catabolic signals. PMID- 17414511 TI - Flexible robotics: a new paradigm. AB - PURPOSE OF REVIEW: The use of robotics in urologic surgery has seen exponential growth over the last 5 years. Existing surgical robots operate rigid instruments on the master/slave principle and currently allow extraluminal manipulations and surgical procedures. Flexible robotics is an entirely novel paradigm. This article explores the potential of flexible robotic platforms that could permit endoluminal and transluminal surgery in the future. RECENT FINDINGS: Computerized catheter-control systems are being developed primarily for cardiac applications. This development is driven by the need for precise positioning and manipulation of the catheter tip in the three-dimensional cardiovascular space. Such systems employ either remote navigation in a magnetic field or a computer-controlled electromechanical flexible robotic system. We have adapted this robotic system for flexible ureteropyeloscopy and have to date completed the initial porcine studies. SUMMARY: Flexible robotics is on the horizon. It has potential for improved scope-tip precision, superior operative ergonomics, and reduced occupational radiation exposure. In the near future, in urology, we believe that it holds promise for endoluminal therapeutic ureterorenoscopy. Looking further ahead, within the next 3-5 years, it could enable transluminal surgery. PMID- 17414512 TI - The evolving therapeutic landscape of prostate cancer. PMID- 17414513 TI - Prostate cancer prevention. AB - PURPOSE OF REVIEW: Prostate cancer, a common cause of morbidity and mortality in the developed world, ought to be a preventable disease. This review focuses on prostate cancer prevention in the context of new mechanistic insights into human prostatic carcinogenesis. RECENT FINDINGS: Evidence is accumulating to implicate infection and inflammation as contributors to prostate cancer development. Inherited prostate cancer susceptibility genes discovered thus far encode participants in host responses to infection. Proliferative inflammatory atrophy, a prostate cancer precursor lesion, ties inflammatory responses to prostatic carcinogenesis. Somatic epigenetic alterations, present in all prostate cancers, appear to arise in the setting of inflammation. Finally, a newly identified somatic genome change, involving a fusion between an androgen-regulated gene, TMPRSS2, and genes encoding members of the ETS family of transcription factors, may provide a clue as to why prostate cancer cells exhibit androgen dependence for growth and survival. SUMMARY: The contributions of infection and inflammation to the early development of prostate cancer suggest prevention strategies featuring prevention or eradication of infection, amelioration of inflammation, or attenuation of genome-damaging reactive oxygen and nitrogen species. The acquisition of androgen dependence later during prostate cancer pathogenesis suggests the use of approaches targeting androgen signaling, including inhibitors of 5alpha-reductase. PMID- 17414514 TI - Dietary intervention in the management of prostate cancer. AB - PURPOSE OF REVIEW: This review integrates recent reports related to the dietary management of prostate cancer with the existing body of science in an effort to best inform practicing clinicians. RECENT FINDINGS: Dietary factors are hypothesized to play a significant role in prostate cancer, and have proven to be important in managing prevalent comorbidities in this patient population (cardiovascular disease, diabetes, and osteoporosis). Data regarding diet and prostate cancer are accumulating and randomized controlled trials are underway which will ultimately yield evidence on which to base recommendations regarding dietary regimens, functional foods, and supplement use. Until that time, most data derive from epidemiologic studies that have limitations in showing cause and effect. During the past year, the greatest and most consistent strides have been made in the area of energy balance, with data consistently showing that overweight and obesity are associated with progressive disease and increased overall mortality. SUMMARY: To date, the strongest evidence regarding diet and prostate cancer relates to energy balance. Urologists aspiring to best clinical practice should encourage their patients to achieve a healthful body weight through regular exercise and a healthful plant-based diet rich in fruits, vegetables, and whole grains. Advocating functional foods or supplements explicitly for cancer control purposes would currently be premature. PMID- 17414515 TI - Prostate cancer immunotherapy. AB - PURPOSE OF REVIEW: Basic immunology research over several decades has led to an improved understanding of tumor recognition by components of the immune system and mechanisms of tumor evasion from immune detection. These findings have ultimately led to four phase III trials, currently underway, evaluating antitumor active immunotherapies in patients with prostate cancer. This article reviews recent published findings in the area of prostate cancer immunotherapies, focusing on both passive and active immunotherapy approaches that have entered clinical trials. RECENT FINDINGS: Clinical trials with immune active agents reported in the last year have demonstrated efficacy in the treatment of prostate cancer. These agents include immune modulators such as granulocyte-macrophage colony stimulating factor and anticytotoxic T-lymphocyte-associated antigen 4 monoclonal antibodies, antibody therapies targeting prostate-specific membrane antigen, and vaccines such as those targeting prostatic acid phosphatase, prostate-specific antigen, and cellular vaccines expressing granulocyte macrophage colony stimulating factor. SUMMARY: Results from several recent clinical trials have suggested that immune-based therapies have clinical benefit in patients with prostate cancer with potentially less toxicity than traditional systemic treatments. We review recent reports of immunotherapies being evaluated in patients with prostate cancer, and highlight the direction for these therapies in combination with other immunotherapies and other traditional therapies. PMID- 17414516 TI - Novel targets and approaches in advanced prostate cancer. AB - PURPOSE OF REVIEW: The development of therapeutic resistance is the underlying cause for most cancer deaths. By understanding the molecular basis of resistance to androgen withdrawal and chemotherapy in prostate cancer, the rational design of targeted therapeutics is possible. We review new treatment options for men with advanced prostate cancer. RECENT FINDINGS: Although the taxanes currently represent the most active chemotherapeutic agents and standard of care for first line treatment of metastatic hormone-refractory prostate cancer, most patients eventually progress because of intrinsic or acquired drug resistance. In recent years, increased knowledge of cancer progression and therapeutic resistance has identified many gene targets that regulate apoptosis, proliferation, and cell signalling. To date, numerous novel compounds have entered clinical trials as either single agents or in combination with cytotoxic chemotherapy. SUMMARY: Even though hormone-refractory prostate cancer is still incurable, it is not untreatable. As cancer cells are proficient at adapting to therapeutic stressors, a combination regimen with drugs that target crucial cellular networks like the apoptotic rheostat may be more promising than treatment with highly selective single-target agents. Recent findings are very hopeful, but challenges remain to demonstrate effective antitumour activity in phase III trials with survival as the principal endpoint. PMID- 17414517 TI - Neoadjuvant therapy and prostate cancer: what a urologist should know. AB - PURPOSE OF REVIEW: Prostate cancer is curable only when treated at an early stage, when the tumor is still localized to the prostate gland. However, even in apparent cases of clinically localized disease, unsuspected extracapsular disease may significantly increase the risk of primary treatment failure. This risk is especially high if the patient has one or more of the following risk factors: a serum prostate-specific antigen level >20 ng/ml, a Gleason score >7, locally advanced disease (clinical stage T3/T4), and extensive disease on prostate biopsy. RECENT FINDINGS: Various regimens of neoadjuvant hormonal therapy and/or chemotherapy have produced mixed results and generally have not influenced the rate of disease relapse (defined by prostate-specific antigen level) in high-risk patients with localized prostate cancer. In addition, antiangiogenic agents, gene therapy, molecular targeting agents, and other promising new therapies have been investigated in a neoadjuvant setting with limited results. SUMMARY: Despite considerable advances, high-risk localized prostate cancer remains an extremely refractory disease. In patients with high-risk prostate cancer, single-modality treatment in the form of surgery offers a 5-year biochemical disease-free survival rate of no better than 50%. To further elucidate optimal treatment regimens for these patients we must actively enrol patients in clinical trials. PMID- 17414518 TI - Energy-based ablative therapy of prostate cancer: high-intensity focused ultrasound and cryoablation. AB - PURPOSE OF REVIEW: This review is an update on the role of the two minimally invasive techniques in primary therapy of organ-confined prostate cancer: as a salvage option after standard-therapy failure and for focal ablation of index tumors. RECENT FINDINGS: Energy-based ablative techniques are of growing interest for today's heterogeneous spectrum of prostate cancer. At present, primary high intensity focused ultrasound appears to be a valid alternative to active surveillance protocols in low-risk cancers and, in older patients, to standard therapy. Morbidity is low, although postoperative impotence occurs frequently. Cryoablation has higher morbidity, even with third-generation conformal technology. With radiorecurrent cancer the potential radiation damage of the rectal wall renders transrectal high-intensity focused ultrasound more hazardous. Third-generation cryoablation seems to give better cancer control with lower morbidity in this situation. Unfortunately, long-term outcome data from controlled trials are not available. SUMMARY: These minimally invasive techniques are not magic bullets, and patients must be informed accordingly. Focal ablation of the prostate segment with the index cancer would minimize morbidity and therefore appears highly appealing. Its success depends on correct localization of the lesion. Until this is achieved with sufficient reliability by appropriate biopsy or imaging techniques it remains strictly experimental. PMID- 17414520 TI - Bibliography. Current world literature. Prostate cancer. PMID- 17414519 TI - Evidence-based clinical practice guidelines for prostate cancer: the need for a unified approach. AB - PURPOSE OF REVIEW: Clinical practice guidelines are being increasingly recognized as critically important to an evidence-based practice. This article reviews the different approaches used by leading urological organizations to the development of prostate cancer guidelines. It further introduces the recommendations of the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) working group as a unified approach to guideline development. RECENT FINDINGS: Clinical guidelines on the management of prostate cancer demonstrate major methodological differences. Most notably, considerable discrepancies with regards to the systems used to grade the quality of the evidence and the strength of recommendation exist. The GRADE approach classifies the quality of evidence as high, moderate, low or very low, according to factors that include study design and execution, and the consistency of the results. It subsequently classifies recommendations as strong or weak, according to the balance between benefits and downsides and the degree of confidence in estimates of the downsides. SUMMARY: There is an urgent need to standardize processes used to develop clinical guidelines for the management of patients with prostate cancer by leading urological organizations. Adoption of the GRADE approach would offer considerable rewards in terms of efficiency, guideline credibility and optimal clinical decision-making. PMID- 17414521 TI - Quantitative X-ray imaging of intraplaque hemorrhage in aortas of apoE(-/-)/LDL( /-) double knockout mice. AB - OBJECTIVES: To determine if hemorrhage into an arterial wall can be detected in CT images by virtue of the iron content. MATERIALS AND METHODS: Aortas from male apoE(-/-)/LDL(-/-) mice (n = 31) were infused in situ with contrast agent, for micro-CT scanning and histology. Roentgen-opacities within the aortic walls were identified by histology and micro-x-ray fluorescence to be iron or calcium. Dual energy scanning was performed at 2 energy levels using synchrotron-based micro-CT [(2 microm)(3) voxels, 16 and 20 keV] and 64-slice CT (0.4 x 0.4 x 0.6 mm voxels, 80 and 120 kVp). RESULTS: Opacities were identified as hemorrhage-related clusters of multiple punctate deposits, containing both Fe (0.48 x 10(-12) g/voxel) and Ca (3.18 x 10(-2) g/voxel), or as isolated confluent accumulations of exclusively calcium. Subtraction of the dual-energy CT scans discriminated iron from calcium deposits. CONCLUSION: Detection and quantification of iron deposits in hemorrhaged atherosclerotic lesions is feasible by dual-energy CT imaging. PMID- 17414522 TI - Evaluation of porcine myocardial microvascular permeability and fractional vascular volume using 64-slice helical computed tomography (CT). AB - OBJECTIVES: Myocardial microvascular permeability-surface area product (MPSP) and fractional vascular volume (FVV), indices of endothelial function and microvascular perfusion, can be noninvasively evaluated by electron beam computed tomography (EBCT), but it remains unknown whether comparable assessments can be obtained with 64-slice multidetector CT (CT-64). METHODS: We studied 12 pigs with both EBCT and CT-64 in randomized order 1 week apart, before and during IV adenosine infusion. Myocardial attenuation changes in the cardiac wall were assessed after a central-venous injection of iopamidol. Time-attenuation curves were analyzed using both indicator-dilution and Patlak models to calculate MPSP and FVV. RESULTS: CT-64 and EBCT assessments of basal MPSP obtained by the Patlak method were similar (0.37 +/- 0.03 vs. 0.37 +/- 0.04 mL/min/g), as was its response to adenosine, and correlated significantly (r = 0.87). Patlak FVV was also similar between CT-64 and EBCT at baseline (0.08 +/- 0.02 vs. 0.07 +/- 0.02 mL blood/mL) and during adenosine, and correlated well (r = 0.93). MPSP and FVV estimated by the indicator-dilution method were not significantly correlated. CONCLUSIONS: CT-64 assessments of myocardial MPSP and FVV may not be reliable when using indicator-dilution analysis, likely due to its sensitivity to scan duration. However, CT-64 assessments obtained using the Patlak model are feasible. PMID- 17414523 TI - Impact of oxygen inhalation on the pulmonary circulation: assessment by magnetic resonance (MR)-perfusion and MR-flow measurements. AB - PURPOSE: Oxygen-enhanced magnetic resonance (MR)-ventilation imaging of the lung is based on the inhalation of a high concentration of oxygen (hyperoxia). However, the effect of hyperoxia on the pulmonary circulation is not yet fully understood. In this study the impact of hyperoxia on the pulmonary circulation was evaluated. MATERIALS AND METHODS: Ten healthy volunteers were examined in a 1.5 T MRI system with contrast-enhanced perfusion MRI (saturation recovery 2D turboFLASH) of the lung and phase-contrast flow measurements in the pulmonary trunk. Both measurements were performed breathing room air (RA) and, subsequently, 100% oxygen (15 L/min) (O(2)). RESULTS: The perfusion measurements showed a significant difference between RA and O(2) for the pulmonary blood flow (181 vs. 257 mL/min/100 mL, P = 0.04) and blood volume (14 vs. 21 mL/100 mL, P = 0.008). The mean transit time of the contrast bolus was not changed (P = 0.4) in the dorsal part of the lung, whereas it was significantly prolonged (P = 0.006) in the central part. The mean heart rate during flow measurements breathing RA (67 +/- 11 beats/min) and O(2) (61 +/- 12 beats/min) were not significantly different (P = 0.055). The average cardiac output (pulmonary trunk) was not significantly lower while breathing O(2) (RA: 5.9 vs. O(2): 5.5 L/min, P = 0.054). CONCLUSION: Hyperoxia causes a significant increase and redistribution of the pulmonary perfusion, whereas it leads to a not significant decrease in cardiac output. Thus, for MR-perfusion and MR-flow measurements oxygen inhalation should be avoided, if possible. In the context of oxygen-enhanced MR-ventilation imaging of the lung the contribution of this effect needs to be further evaluated. PMID- 17414524 TI - Flat panel detector-based volumetric computed tomography (fpVCT): performance evaluation of volumetric methods by using different phantoms in comparison to 64 multislice computed tomography. AB - OBJECTIVES: Evaluation of a silicon-based flat panel volumetric computed tomography (fpVCT) and multislice CT in terms of volumetry of phantoms with different algorithms. Furthermore, to compare the different volumetric analysis methods themselves. MATERIALS AND METHODS: Four phantoms of different materials have been scanned with fpVCT (GE prototype with circular gantry with 2 aSi/CsI flat panel detector) and a 64-slice spiral CT (MSCT: LightSpeed VCT). Three spherical phantoms of different materials and 1 phantom with an irregular shape were evaluated. True volumes were calculated in dependence from the diameter or by water displacement method. Imaging parameters (80 kVp, 100 mA) and the position of the phantoms were identical in both techniques. After reconstruction of the images different algorithms have been used 4 times for each phantom. These analysis methods have been performed: Region growing, threshold method, planimetry, 3-dimensional volumetry measurement by using the equation of an ellipsoid (ellipse) and an advanced lung analysis modus [single advanced lung analysis (ALA)]. The mean values and the standard deviations have been evaluated and compared with the true volumes. RESULTS: In all phantoms fpVCT showed better results with lower deviations from the true values than in MSCT, especially for small volumes of the phantoms. However, the results of the ALA single method demonstrated no significant difference between the fpVCT and MSCT. The comparison of the different analysis methods revealed that 3-dimensional measurement with the ellipse method was the worst method for volume estimation, especially for the irregularly formed phantom. CONCLUSION: fpVCT was superior to MSCT in the volumetry of small objects. The ellipse method has been shown to be the worst for volumetry with the highest relative deviations from the true volume value. The single ALA method shows the lowest standard deviation thereby revealing a reproducible volumetric method for small nodules. However, further future developments of volumetric analysis methods are necessary to use them accurately in daily routine. Due to the truly isotropic volume data set with high spatial resolution fpVCT is a powerful tool for the volumetry of small nodules. PMID- 17414525 TI - Automated volumetry of solid pulmonary nodules in a phantom: accuracy across different CT scanner technologies. AB - OBJECTIVES: The accuracy of automated volumetry for pulmonary nodules in a phantom using different CT scanner technologies from single-slice spiral CT (SSCT) to 64-slice multidetector-row CT (MDCT) was compared. MATERIALS AND METHODS: A lung phantom with 5 different categories of pulmonary nodules was scanned using a single-slice spiral CT, a 4-slice MDCT, a 16-slice MDCT and a 64 slice MDCT. Each category comprised of 7-9 nodules each (total n = 40) with different known volumes. Standard dose and low dose protocols were performed using thin and thick collimation. Image data were reconstructed at the thinnest slice thickness. Data sets were analyzed with a dedicated volumetry software. Volumes of all nodules were calculated and compared. RESULTS: Mean absolute percentage error (APE) for all nodules was 8.65% (+/-7.29%) for the SSCT, 10.26% (+/-8.25%) for the 4-slice MDCT, 8.19% (+/-7.57%) for the 16-slice MDCT and 7.89% (+/-7.39%) for the 64-slice MDCT. There was statistically significant influence of the scanner type, protocol, anatomic location, and nodule volume on APE, but overall, APEs were comparable. CONCLUSION: Computer-aided volumetry showed accurate measurements in all tested scanner types. This finding has important implications for nodule assessment and follow-up. PMID- 17414526 TI - The efficacy of nanoscale poly[bis(trifluoroethoxy) phosphazene] (PTFEP) coatings in reducing thrombogenicity and late in-stent stenosis in a porcine coronary artery model. AB - OBJECTIVES: Previous experimental studies have demonstrated that poly[bis(trifluoroethoxy)phosphazene] (PTFEP) nanocoated stents have antithrombotic characteristics, reduce in-stent stenosis, prevent wall inflammation, and do not hamper endothelialization. This study was designed to validate these findings in a porcine coronary artery model. MATERIALS AND METHODS: PTFEP-coated (n = 15) and bare stents (n= 13) were implanted in coronary arteries of 18 mini-pigs (4- and 12-week follow-up). Primary study endpoints were thrombogenicity and in-stent stenosis, secondary study endpoints were inflammatory response and re-endothelialization evaluated by quantitative angiography and light microscopy. RESULTS: No thrombus deposition occurred on any stent. At 4 weeks follow-up, the bare stents (n = 4) had a significantly smaller neointimal area (1.93 vs. 3.20 mm(2), P = 0.009). At 12 weeks, PTFEP-coated stents (n = 11) had significantly superior results in almost all parameters: neointimal area (2.25 vs. 2.65 mm(2), P = 0.034), neointimal height (204.46 vs. 299.41 microm, P = 0.048), percentage stenosis (38.25 vs. 50.42%, P = 0.019), and inflammation score (0.12 vs. 0.30, P = 0.029). Complete re-endothelialization was seen in both stent types at both intervals. CONCLUSION: At long-term follow-up, the superior results of PTFEP-coated stents were characterized by a noteworthy reduction of neointimal growth and inflammatory response. PMID- 17414527 TI - Sonographic findings in a model of ischemia-induced necrotizing enterocolitis with pathological correlations. AB - PURPOSE: To evaluate sonographic findings in ischemic enterocolitis (IEC) and correlate with pathologic findings in an experimental study. MATERIALS AND METHODS: Ischemic enterocolitis was induced with ligation of the superior mesenteric artery in 20 rabbits. Plain radiography and ultrasonography (US) were performed. US was done hourly after the ligation using 10 MHz linear probe. US findings were categorized into 2 groups according to the bowel wall echogenicity; the echogenic dots (ED) group and the circumferential granular echogenicity (CGE) group. US findings were compared with the specimen radiography and the histopathology. RESULTS: On US, ED were seen in the bowel of all rabbits after SMA ligation (2.2 +/- 1.3 hours [standard deviation]) and CGE in 16 rabbits (4.1 +/- 0.9 hours). On the specimen radiographs, multiple radiolucent air bubbles were present. Comparing the ED and CGE group, histopathological findings revealed the CGE group had severer injury of the bowel wall than the ED group. On plain radiography, there was progressive bowel distention, but pneumatosis intestinalis (PI) was not evident. CONCLUSION: ED or CGE are the sonographic findings of ischemic enterocolitis, and bowel wall echogenicity might reflect the degree of ischemic injury. PMID- 17414528 TI - Contrast-to-noise ratios of different elements in digital mammography: evaluation of their potential as new contrast agents. AB - PURPOSE: To determine the contrast-to-noise ratios (CNRs) of different elements at different energies using various anode/filter combinations currently employed in digital mammography. The elements investigated included not only elements already used in conventional contrast agents such as gadolinium and iodine but also other elements to investigate their potential as mammographic contrast agents. MATERIALS AND METHODS: The CNRs of 20 mmol/L bismuth (Bi), gadolinium (Gd), ytterbium (Yb), dysprosium (Dy), and iodine (I) were determined at different slice thicknesses (0.25, 0.5, and 1 cm) of the element solution with an additional 4-cm Plexiglas in relation to water (to simulate dense glandular tissue), oil, and air. The following anode/filter combinations were used: Mo/Mo in the range of 22-34 kVp, Mo/Rh in the range of 36-40 kVp, Rh/Rh in the range of 42-46 kVp, and Mo/Cu in the range of 47-49 kVp. In the range of 22-46 kVp, the mAs were chosen to achieve a fairly uniform dose range (of 4.38-4.71 mGy). Doses were measured using the PTW DIADOS diagnostic dosimeter. The element solutions were examined with a GE Senographe 2000D. RESULTS: Bismuth showed the best CNR for all energies investigated and in relation to both water and oil. In the energy range below 46 kVp, bismuth (CNR at 30 kVp/50 mAs and 1/0.5/0.25 cm slice thickness: 9.9/6.1/3.4) was followed by Yb (5.9/3.5/2.0), Dy (5.3/3.2/1.9), Gd (4.2/2.5/1.6), and iodine (2.4/1.8/1.5). Bismuth had the best CNR relative to both water (values given above) and oil (Bi: 20.7/11.2/5; Yb: 16.9/8.6/3.6; Dy: 16.6/8.4/3.5; Gd: 15.21/7.5/3.2; I: 13.8/6.3/3.2). The CNR of Bi was also superior to that of the other elements investigated at high energy in combination with copper filters (eg, CNR at 49 kVp Mo/Cu at slice thicknesses of 1/0.5/0.25 cm, relative to water: 9.6/6.0/4.0) but now followed by iodine (7.9/5.3/3.5), Yb (5.8/4.0/2.9), Dy (5.4/3.7/2.8), and Gd (4.7/3.2/2.7). Iodine was the only element of those investigated whose contrast-to-noise ratio was improved with the use of a copper filter at high energies based on its K-edge (increase in CNR from 2.9 to 7.9 from 40 to 49 kVp at 1-cm slice thickness). Nevertheless, the improved CNR of iodine was below that of Bi at low energies and for Mo/Mo or Mo/Rh filters. The contrast of water/fat tended to decrease slightly at higher energies (CNR of water/air at 42 kVp: 33.9, at 48 kVp: 25.6; CNR of oil/air at 42 kVp: 23.8, at 48 kVp: 21.9). CONCLUSION: Copper filters and higher energies are useful for visualizing iodine-based contrast agents in contrast-enhanced mammography because they markedly improve the CNR relative to water. This technique further benefits from the fact that the CNR of water and fat relative to air markedly decreases at higher energies and with the use of copper filters. Bismuth was found to have a much better CNR than iodine for all energies investigated including the low energy ranges typically used in mammography. These results suggest that bismuth is a potential candidate for a specific mammographic contrast agent. PMID- 17414529 TI - Awareness and perceptions of fibromyalgia syndrome: a survey of southeast asian rheumatologists. AB - BACKGROUND: Fibromyalgia syndrome (FMS) is a common but controversial condition. There appears to be different level of belief of its existence and awareness. We set out to explore the variations of perceptions and awareness of this condition among rheumatologists from the Southeast Asia (SEA) region. METHODS: One hundred eight rheumatologists from the participating countries; 28 from Malaysia, 20 from Singapore, 26 from Thailand, 2 from Brunei, and 42 from Indonesia were approached to participate in this survey by answering specific questions regarding their beliefs in relation to FMS; 82% respondents from Malaysia, 100% from Singapore, 92% from Thailand, 100% from Brunei, and 90% from Indonesia completed the questionnaires. RESULTS: Most rheumatologists (92.5%) from SEA believe that FMS is a distinct clinical entity, and also this condition is considered an illness rather than a disease. Eighty-seven percent rheumatologists from SEA believe that FMS is a mixture of medical and psychological illness, 9% believe that FMS is primarily a psychological illness, and 3% believe that it is a medical illness. Only 60% of those in a university setting include FMS in their undergraduate teaching. Eighty-five percent of the respondents ordered blood tests to exclude other serious pathologic conditions, and 100% of the respondents from SEA countries also prescribed some form of drugs to FMS patients. CONCLUSION: FMS is apparently seen worldwide. This study confirmed that there was a variation of perceptions and knowledge of FMS among rheumatologists from SEA countries. However, most rheumatologists agreed that FMS is a distinct clinical entity with a mixture of medical and psychological factors. PMID- 17414530 TI - Risk factors for pes anserinus tendinitis/bursitis syndrome: a case control study. AB - BACKGROUND: Pes anserinus tendinitis/bursitis (PATB) is a frequent cause of knee pain. Its predisposing factors are still controversial. OBJECTIVES: Assess the effect of a set of demographic, clinical, somatometric, and biomechanical factors on the risk for PATB. METHODS: A case control design was used to evaluate the association between clinically diagnosed PATB and the presence of diabetes; knee osteoarthritis (and its radiographic severity); obesity; knee collateral, and anteroposterior instability; and knee or hindfoot malalignment. RESULTS: Twenty two consecutive, incident PATB patients were included; all were females 62.1 +/- 11.5-year-old (limits 45-82). Thirty-eight sex- and age-matched (59.8 +/- 9.4 year-old; P = 0.41) subjects were used as controls; these had asymptomatic osteoporosis (20) and a series of rheumatic syndromes (18). There was no difference in prevalence of diabetes, knee osteoarthritis, obesity, knee instability, varus knee deformity, and hindfoot malalignment between cases and controls. Furthermore, no difference in overall, lateral, medial, and patellofemoral knee osteoarthritis radiographic severity mean score was found between study groups. The presence of valgus knee deformity alone (OR: 5.2; 95% CI: 1.1-25.5), or in combination with collateral instability (OR: 6.0; 95% CI: 1.4-26.0), was identified as associated with PATB. CONCLUSIONS: Valgus knee deformity, alone or in association with collateral instability, seems to be a risk factor for PATB. No association was found between PATB and some conditions previously reported as predisposing factors such as diabetes, knee osteoarthritis, and obesity. PATB should be kept in mind as a highly probable diagnosis in mature women with medial knee pain and valgus knee deformity. PMID- 17414531 TI - Rheumatoid arthritis patients' experiences of night pain. AB - BACKGROUND: Night pain is a significant problem for many patients with rheumatoid arthritis (RA), but clinicians often overlook it. This study aimed to explore the issue of night pain among patients with RA both at home and in the hospital setting. METHODS: This was a qualitative study involving in-depth interviews among 8 hospital inpatients with established RA. Each interview was recorded, transcribed, and underwent framework analysis. RESULTS: The following themes were identified: Night pain in RA is a significant problem both at home and in hospital. All patients had established routines that they used in the home setting when disturbed by night pain. This often included getting out of bed and walking around, making hot drinks, and/or taking extra painkillers. These activities were often curtailed in the hospital setting for fear of disturbing other patients or the nurses. The effect of disturbed sleep and night pain on other people, be it partners or family members at home or patients and nurses in the hospital, was raised by all interviewees. CONCLUSION: Night pain in particular appears harder to control in the hospital setting than at home, and some of this may stem from lack of a tailored routine and fears of disturbing others. Information could be provided when patients are first admitted to the ward. This could describe or orient patients to the facilities available, which may be beneficial in the management of night pain. Further work on identifying influences upon night pain needs to be undertaken. PMID- 17414532 TI - Implementation of a mandatory rheumatology osteoporosis consultation in patients with low-impact hip fracture. AB - BACKGROUND: Osteoporosis remains an underdiagnosed and undertreated major health problem. The current treatment rate for patients who have experienced at least 1 osteoporotic fracture is 20%-25%. Therefore, the Rheumatology and Internal Medicine Departments of Ochsner Clinic Foundation New Orleans implemented a mandatory rheumatology osteoporosis consult as part of preprinted admission orders for all patients after hip fracture surgery on the Internal Medicine service. METHODS: We conducted a retrospective study of 78 patients admitted with a low-impact hip fracture between June 2004 and July 2005. These patients were seen by the rheumatology service in the hospital after hip fracture repair (exposed group). Osteoporosis evaluation was performed based on an interview questionnaire. Seventy-eight age-matched patients previously admitted for low intensity or low-impact hip fracture in 2002-2003 but not exposed to the mandatory rheumatology consult served as our comparison group. Pearson chi2 test was used for statistical analysis. RESULTS: Mean patient age was 80 years. Of the 78 unexposed patients, 17 (22%) were on treatment (calcium, vitamin D, hormones or antiresorptive agents) before the hip fracture, and 18 (23%) were on treatment after fracture repair. Of the 78 patients exposed to the compulsory rheumatology consultation, 34 (44%) patients were receiving osteoporosis treatment before hip fracture and 75 (96%) patients were receiving treatment after fracture repair. Of the patients not treated before hip fracture repair, there was a significant increase in the percent treated for those patients exposed to the rheumatology consult versus those not exposed (97.6% vs. 2.4%, respectively, P < 0.0001). CONCLUSIONS: In our institution, we were successful in identifying and initiating appropriate therapy for osteoporosis patients through an automatic rheumatology osteoporosis consultation after hip fracture. The implementation of a mandatory osteoporosis consult resulted in a statistically significant increase in treatment of the exposed group compared with the unexposed group. PMID- 17414533 TI - Risk factors associated with the cumulative survival of low-dose methotrexate in 273 Japanese patients with rheumatoid arthritis. AB - BACKGROUND: Methotrexate (MTX) has become the most commonly prescribed disease modifying antirheumatic drug (DMARD) for the treatment of rheumatoid arthritis (RA). In the Western countries, the MTX dosage is safely increased to a maximum of 25 mg/wk until a significant response is observed. On the contrary, in Japan, MTX has been approved only as a second-line agent, and the approved maximum MTX dosage is only 8 mg/wk. This suboptimal dosage may affect MTX survival in Japanese RA patients. OBJECTIVES: To study risk factors associated with the cumulative survival of MTX in Japanese RA patients. METHODS: Data on 273 patients (male 44, female 229) with RA treated with MTX between January 1, 2000 and September 30, 2004 in our center were studied. RESULTS: Two hundred seventy-three patients were followed for 437 person-years of MTX exposure. Mean MTX dosage was 5.5 +/- 1.9 mg/wk. The cumulative MTX survival probability after 5 years was 61.9%. Univariate Cox regression analysis showed a significant correlation between MTX survival probability and use of fewer previous DMARDs, higher dose of MTX, inclusion of folate supplementation, and shorter disease duration. In the multivariate Cox regression model, use of fewer previous DMARDs remained significantly related to MTX survival. Reasons for discontinuation included adverse effects in 34 patients (12.5%) and inefficacy in 6 patients (2.2%). CONCLUSIONS: Cumulative survival was the same or slightly better than those in reports from Western countries, with less withdrawals reported due to adverse events or inefficacy. Whether these results are due to different MTX needs in Japanese or to acceptance of less clinical efficacy will require further studies. The use of fewer previous DMARDs was associated with longer MTX survival. PMID- 17414534 TI - Successful etanercept use in an HIV-positive patient with rheumatoid arthritis. AB - Limited information exists on treatment of rheumatoid arthritis in a patient with coexisting human immunodeficiency virus (HIV) infection. We report a case of a patient with rheumatoid arthritis who then became HIV positive. His HIV viral load was controlled with antiretroviral therapy, but he continued to have active rheumatoid arthritis despite therapy with hydroxychloroquine, sulfasalazine, and corticosteroids. Because of unremitting rheumatoid disease, we are now treating him with a TNFalpha inhibitor, and his rheumatoid disease activity has decreased from 28 swollen and tender joint count to less than 5. PMID- 17414535 TI - Minor neck trauma in chronic ankylosing spondylitis: a potentially fatal combination. AB - Long-standing ankylosing spondylitis may predispose a patient to serious cervical injury in the setting of minor trauma. Early diagnosis is essential to a favorable outcome. We report a 75-year-old man whose relatively minor trauma in the setting of AS resulted in a cervical fracture and callus formation, which masqueraded as a tumor. The patient developed neck pain, bilateral hypoglossal nerve palsy with dysarthria, and dysphagia that ultimately resulted in his death. This case illustrates progressive neurologic signs of gradual disarticulation of the skull from the cervical spine. The situation is considered of importance because it emphasizes the need for early recognition and possible intervention in the presence of hypoglossal symptoms. The specific combination of long-standing ankylosing spondylitis and minor trauma is one setting in which a clinician must be alerted. Early consideration of neck immobilization is emphasized. PMID- 17414536 TI - Lupoid sclerosis: what is the role of antiphospholipid antibodies? AB - A multiple sclerosis-like syndrome, also called lupoid sclerosis for having some lupus characteristics, has been described in a few reports. Recently, antiphospholipid antibodies have been demonstrated in patients with this syndrome, suggesting that they can participate in the etiopathogenic process, which can have therapeutic implications. We report the case of a patient previously diagnosed with multiple sclerosis who later presented with ANA positivity and antiphospholipid antibodies, livedo reticularis, and deep venous thrombosis, leading her to be characterized as having lupoid sclerosis. The patient was anticoagulated. Antiphospholipid antibodies may be involved in the pathogenesis of lupoid sclerosis. PMID- 17414537 TI - Piriformis pyomyositis with sciatica: an unrecognized complication of "unsafe" abortions. AB - We report the case of a patient who presented with right sciatic pain as a manifestation of piriformis muscle syndrome 1 month after a clandestine- induced abortion. MRI revealed the presence of a piriformis abscess. Staphylococcus aureus was isolated from blood cultures and from the purulent material obtained by CT-guided aspiration. Piriformis pyomyositis should be recognized as a potential complication of "unsafe" abortions that if left untreated could progress to sepsis and death. PMID- 17414538 TI - Successful rituximab therapy of acquired factor VIII inhibitor in a patient with rheumatoid arthritis. AB - Acquired factor VIII deficiency due to antibody inhibition can result in life threatening hemorrhage. Rarely such antibody inhibition of factor VIII can be associated with other autoimmune disorders including rheumatoid arthritis. We present the first case of a patient with active rheumatoid arthritis and refractory bleeding diatheses due to a factor VIII inhibitor who was successfully treated with rituximab. A 61-year-old Caucasian female with rheumatoid arthritis unresponsive to multiple therapies developed an acute hematoma while having a peripheral catheter placed. Her aPTT was prolonged at 61.4 with low factor VIII activity and an inhibitor level for factor VIII of 2.0 Bethesda Units. She received rituximab 375 mg/m in 4 weekly doses. Normalization of the aPTT and resolution of the bleeding occurred in 2 weeks. After 45 days, the levels of factor VIII inhibitor and factor VIII activity were <0.4 BU/mL and 130%, respectively. After 1 year, the aPTT remained normal and there was no further bleeding. An added benefit was the substantial improvement in her rheumatoid arthritis. Treatment of acquired factor VIII inhibitors in rheumatoid arthritis should be guided by the levels of the inhibitor. Patients with low levels of the inhibitor may respond to rituximab monotherapy, whereas higher levels may necessitate combination therapies. The dual benefit of RA disease control and resolution of bleeding makes rituximab therapy compelling in the rare patient who presents with these 2 disorders. PMID- 17414539 TI - Relapsing splenic vein thrombosis associated with antiphospholipid antibodies in a patient with wegener granulomatosis. AB - Relapsing splenic vein thrombosis, a very rare complication of Wegener granulomatosis (WG), is described in a female patient. Positive antiphospholipid antibodies found in this case are a rare occurrence in primary vasculitis, especially in WG. This probably caused or accentuated an effect of the WG on the splenic vein. Treatment of such patients with antiphospholipid syndrome and WG must include anticoagulation and immunosuppression and, as noted in this patient, the splenic vein thrombosis and other antiphospholipid syndrome and WG can resolve quickly. PMID- 17414540 TI - Development of serum sickness-like symptoms after rituximab infusion in two patients with severe hypergammaglobulinemia. AB - Rituximab is a monoclonal antibody that depletes B cells and is commonly used for the treatment of autoimmune diseases. In this report, we describe 2 patients with autoimmune diseases and marked hypergammaglobulinemia attending at a tertiary care hospital, who, following treatment with rituximab, developed a severe serum sickness-like reactions. Both patients were treated with intravenous corticosteroids, recovered well, and on follow-up, were negative for the development of human antichimeric antibodies. Based on these and other similar cases in the literature, we propose that severe hypergammaglobulinemia due to autoimmune conditions may be a predisposing factor for the development of a serum sickness-like reaction after treatment with Rituximab, which can sometimes occur without prior exposure to this antibody. Rheumatologists should be aware of this possible association when prescribing this medication. PMID- 17414541 TI - Basic science for the clinician 43: the mitogen-activated protein kinase family in inflammatory signaling. AB - Growth factors, cytokines, chemokines, apoptotic stimuli, TLR ligands, stress-a lot can happen to a cell in day-to-day living, but these are extracellular phenomena, and all influence the inner workings of the cell. Once the ligand binds to the receptor, or the stress, occurs, changes occur within the cell that eventuate in intranuclear transcription of new genes, subsequent changes in cellular structure and function, and often in the release of chemicals that transmit signals to self (autocrine) and other cells (paracrine and juxtacrine). Often, these changes occur due to the activation of cascades of kinases, a perireceptor choreography that is only now being elucidated. The human "kinome" (the various families of kinases) is very complex; we will limit ourselves in this discussion to the mitogen-activated protein kinases. PMID- 17414542 TI - Don't forget the pets! PMID- 17414543 TI - Fibromyalgia: update on mechanisms and management. PMID- 17414545 TI - Digital densitometry for evaluation of osteoporosis in spondyloartropathy patients. PMID- 17414546 TI - Cutaneous hyperpigmentation during therapy with hydroxychloroquine. PMID- 17414547 TI - More on polymyalgia rheumatica (PMR) as a paraneoplastic rheumatic syndrome in the elderly (bicytopenia and PMR preceding acute myeloid leukemia). PMID- 17414548 TI - Preinjury status: are orthopaedic trauma patients different than the general population? AB - OBJECTIVE: To describe the preinjury health-related quality of life (HRQL) of orthopaedic trauma patients admitted to Level I trauma centers relative to the general population. DESIGN: Prospective cohort study using retrospectively collected preinjury HRQL measures. SETTING: Two Level I adult trauma centers in Melbourne, Australia. PARTICIPANTS: A total of 2388 admitted orthopaedic trauma patients aged > or = 18 years were included, captured by the Victorian Orthopaedic Trauma Outcomes Registry (VOTOR) between October 2003 and January 2006. Patients with a significant head injury (Abbreviated Injury Scale severity score >2), dementia, mental illness, mental disability, who were non-English speakers, or who were postoperatively confused were excluded. MAIN OUTCOME MEASUREMENTS: The 12-item Short Form Health Survey (SF-12). RESULTS: The preinjury SF-12 was obtained for 1839 patients [median (interquartile range) of 6 (3-12) days postinjury]. The VOTOR population reported mean physical SF-12 scores above population norms (50.9 vs. 48.9, P < 0.001). The differences were predominant in men and confined to patients aged 18 to 54 years. The mean mental SF-12 scores of the VOTOR population were also greater than population norms (54.5 vs. 52.4, P < 0.001) and this was most apparent for women. CONCLUSIONS: Establishing the preinjury HRQL of trauma patients is important for evaluating the quality of orthopaedic trauma patient outcomes. Reliance on population norms for this purpose may lead to an underestimation of the impact of injury in particular age and sex subgroups, but given the size of the differences seen, any underestimation would be small. Where individual preinjury data cannot be collected, population norms could be used as a reasonable estimate of preinjury patient status for assessing quality of recovery. PMID- 17414549 TI - Avascular necrosis of the femoral head in pertrochanteric fractures: a report of 8 cases and a review of the literature. AB - OBJECTIVE: To evaluate patients with pertrochanteric fractures (Type 31A1 + 2 ASIF/OTA classification) treated surgically and who subsequently developed avascular necrosis of the femoral head (ANFH). SETTING: University hospital. DESIGN: Retrospective clinical study and analysis of the literature. PATIENTS: From 1995 to 2003 the authors operated on 1,373 patients with pertrochanteric fractures. Eight patients with a mean age of 69 years (range, 52-78 years) subsequently developed ANFH. INTERVENTION: Five patients were treated with a dynamic hip screw (DHS), and 3 were treated with an intramedullary hip nail (IMHN). RESULTS: All fractures in these 8 patients healed without complications in 3 to 4 months. Avascular necrosis of the femoral head developed 4 months to 4 years after the operation. Detailed analysis of these cases did not reveal any risk factors or errors in surgical technique to account for the ANFH. Results of a literature review of 41 cases in addition to our 8 cases give us reason to believe the etiology of postoperative AFNH following surgical treatment of pertrochanteric fractures is probable injury to the deep branch of the medial circumflex artery at time of fracture. Other possible risk factors include high energy trauma, comminution, displacement of the fragments, and atypical fracture line. CONCLUSIONS: ANFH develops in approximately 0.5-1% of all pertrochanteric fractures, mostly within 4 years after the injury and predominantly in patients older than 50 years of age. The diagnosis should be considered in patients who have undergone an operation previously and have symptoms that are unclear. Prevention of ANFH in these fractures includes an early reduction, stable fixation, and correct surgical technique. PMID- 17414550 TI - Modified Pauwels' intertrochanteric osteotomy in neglected femoral neck fractures in children: a report of 10 cases followed for a minimum of 5 years. AB - OBJECTIVES: To evaluate the role of a modified Pauwels' intertrochanteric osteotomy (MPIO) in neglected femoral neck fractures in children. DESIGN: Prospective study with retrospective analysis. SETTING: Tertiary care Postgraduate Institute of Medical Sciences. PATIENTS: Ten children (8 males, 2 females) with an average age of 10.2 years with neglected femoral neck fractures were seen from 1990 to 1998. A femoral neck fracture was considered neglected when no proper medical treatment was instituted for at least 1 month following the fracture. Nonunion was accompanied by coxa vara and resorption of the femoral neck in 9 patients; a 10th patient had a neglected femoral neck fracture for 1 month without coxa vara. Three patients at time of presentation with Delbet Type II displaced fractures with associated nonunion and coxa vara (2 with Ratliff Type III and 1 with Type I) also had avascular necrosis using plain radiographic criteria of increased density. INTERVENTION: Modified Pauwels' intertrochanteric osteotomy. The children were immobilized in a hip spica for 6-10 weeks postoperatively and weightbearing was started after hip spica removal. MAIN OUTCOME MEASUREMENTS: Fracture healing, neck-shaft angle, avascular necrosis, and functional outcome. RESULTS: Patients were followed for an average of 8.2 years (range 5-12 years). All patients had union of their fracture within an average of 16.6 weeks (12-20 weeks) and of the osteotomy site within 8.2 weeks (7-9 weeks). Radiologic signs of avascular necrosis disappeared completely in the 3 patients who presented with avascular necrosis. In 1 patient with a preoperatively viable femoral head, radiologic signs of Ratliff Type I avascular necrosis appeared between 60 and 98 weeks. This radiologic finding became normal again, indicating viability of the femoral head somewhere between 98 to 205 weeks of follow-up. Postoperatively, an average of 135-degree neck-shaft angle was achieved (range 125-160 degrees). The average preoperative neck-shaft angle was 104.4 degrees (range 92-120 degrees) and on the normal hip side it was 127.7 degrees (range 124 132 degrees). Significant improvement in the neck-shaft angle was seen compared with the preoperative angle (P < 0.001) and normal hip angle (P < 0.05). Coxa vara and signs of chondrolysis were not observed in any of the patients. Premature proximal femoral epiphyseal closure resulting in a 1-cm and a 1.5-cm leg-length discrepancy was seen in 2 patients as compared with their normal side. A mild Trendelenburg gait was observed in 1 patient (10%). Using Ratliff's criteria, 9 patients (90%) were graded as a good result and 1 patient (10%) was graded as a fair result. The osteotomy plate was removed in 1 patient (10%). CONCLUSION: An MPIO creates a biomechanical environment conducive to healing of a neglected femoral neck nonunion in a child while simultaneously correcting an associated coxa vara. The procedure also seems to have a biological role in helping restore viability to a noncollapsed femoral head with avascular necrosis. PMID- 17414551 TI - Does medial tenderness predict deep deltoid ligament incompetence in supination external rotation type ankle fractures? AB - OBJECTIVE: To identify whether medial tenderness is a predictor of deep deltoid ligament incompetence in supination-external rotation ankle fractures. DESIGN: All Weber B lateral malleolar fractures with normal medial clear space over a 9 month period were prospectively included in the study. Fracture patterns not consistent with a supination-external rotation mechanism were excluded. SETTING: High-volume tertiary care referral center and Level I trauma center. PATIENTS/PARTICIPANTS: Fifty-five skeletally mature patients with a Weber B lateral malleolar fracture and normal medial clear space presenting to our institution were included. INTERVENTION: All study patients had ankle anteroposterior, lateral, and mortise radiographs. Each patient was seen and evaluated by an orthopedic specialist and the mechanism of injury was recorded. Each patient was assessed for tenderness to palpation in the region of the deltoid ligament and then had an external rotation stress mortise radiograph. MAIN OUTCOME MEASURE: Correlating medial tenderness with deep deltoid competence as measured by stress radiographs. RESULTS: Thirteen patients (23.6%) were tender medially and had a positive external rotation stress radiograph. Thirteen patients (23.6%) were tender medially and had a negative external rotation stress radiograph. Nineteen patients (34.5%) were nontender medially and had a negative external rotation stress radiograph. Ten patients (18.2%) were nontender medially and had a positive external rotation stress radiograph. We calculated a chi statistic of 2.37 as well as the associated P value of 0.12. Medial tenderness as a measure of deep deltoid ligament incompetence had a sensitivity of 57%, a specificity of 59%, a positive predictive value of 50%, a negative predictive value of 66%, and an accuracy of 42%. CONCLUSION: There was no statistical significance between the presence of medial tenderness and deep deltoid ligament incompetence. There is a 25% chance of the fracture in question with medial tenderness having a positive external rotation stress and a 25% chance the fracture with no medial tenderness having a positive stress test. Medial tenderness in a Weber B lateral ankle fracture with a normal clear space on standard plain radiographs does not ensure the presence of a positive external rotation stress test. PMID- 17414552 TI - Methods of operative fixation of the acromio-clavicular joint: a biomechanical comparison. AB - PURPOSE: Three different methods of fixation used in acute disruption of the acromio-clavicular (AC) joint-namely, the coraco-clavicular Bosworth screw (CC Screw), a coraco-clavicular sling of Mersilene #5 tape (CC Sling), and a Hook Plate-were compared to baseline to see which could most closely replicate the stiffness of healthy cadaveric AC specimens (Intact). HYPOTHESIS: It is hypothesized that the Hook Plate method, as compared with the other reconstructions tested, will be most similar mechanically to the intact AC joint with respect to present outcome measures. METHODS: Five matched pairs of fresh frozen cadaveric specimens were tested. Stiffness was tested with superior cyclic loads to 70 N. The stiffness for each specimen was initially tested with all the ligaments in place (Intact). The AC and CC ligaments were then sectioned, and stiffness was tested, in varying order, with reconstructions using the CC Screw, the CC Sling, and the Hook Plate. Failure testing consisted of taking either the CC Screw or Hook Plate to failure within each matched pair. RESULTS: The CC Screw and the CC Sling, respectively, showed stiffnesses of 46 +/- 23 N/mm and 15 +/- 8 N/mm, which was significantly different from the Intact specimen (P < 0.05). The Hook Plate had a stiffness of 26 +/- 17 N/mm, most comparable to the Intact joint stiffness of 25 +/- 8 N/mm (P = 0.785). With failure testing, the CC Screw failed at a significantly higher load than the Hook Plate (744 +/- 184 N vs 459 +/- 188 N) (P = 0.034). CONCLUSION: The CC Screw demonstrated the greatest stiffness with repetitive loading to 70 N. The Hook Plate had a stiffness most similar to the normal physiologic state of the AC joint. The CC Sling was significantly less stiff than the Intact joint or the other methods of fixation. SIGNIFICANCE: Although the stiffest construct is the CC Screw, Hook Plate fixation allows physiologic motion without pathological deformation and most closely resembles the stiffness of the native AC joint for the current test procedure used. PMID- 17414553 TI - Characterization of extremity wounds in Operation Iraqi Freedom and Operation Enduring Freedom. AB - OBJECTIVES: Extremity wounds and fractures traditionally comprise the majority of traumatic injuries in US armed conflicts. Little has been published regarding the extremity wounding patterns and fracture distribution in the current conflicts in Iraq and Afghanistan. The intent of this study was to describe the distribution of extremity fractures during this current conflict. DESIGN: Descriptive epidemiologic study. METHODS: The Joint Theater Trauma Registry was queried for all US service members receiving treatment for wounds (ICD-9 codes 800-960) sustained in Operation Iraqi Freedom (OIF) and Operation Enduring Freedom (OEF) from October 2001 through January 2005. Returned-to-duty and nonbattle injuries were excluded. Wounds were classified according to region and type. Extremity wounds were analyzed in detail and compared to published results from previous conflicts. RESULTS: A total of 1281 soldiers sustained 3575 extremity combat wounds. Fifty-three percent of these were penetrating soft-tissue wounds and 26% were fractures. Of the 915 fractures, 758 (82%) were open fractures. The 915 fractures were evenly distributed between the upper (461, 50%) and lower extremities (454, 50%). The most common fracture in the upper extremity was in the hand (36%) and in the lower extremity was the tibia and fibula (48%). Explosive munitions accounted for 75% of the mechanisms of injury. CONCLUSIONS: The burden of wounds sustained in OIF/OEF is extremity injuries, specifically soft-tissue wounds and fractures. These results are similar to the reported casualties from previous wars. PMID- 17414554 TI - Antibiotic cement-coated interlocking nail for the treatment of infected nonunions and segmental bone defects. AB - Chronic infection of bone with nonunion and/or bone defects is traditionally treated by a 2-stage procedure involving initial debridement and antibiotic delivery and then definitive internal fixation. Alternatively, external fixators are used to provide stability. A technique with which antibiotic cement-coated interlocking intramedullary nails are prepared in the operating room with the use of nails and materials that generally are available is herein described. Although useful for all infected nonunions and/or segmental bone defects, this technique is particularly useful for patients who are not ideal candidates for external fixation and for those who do not want to have an external fixator applied. This technique was used in a series of 20 patients. In 17 patients, the goal of bony union was achieved (85%). In the remaining 3 patients (15%), the goal of control of infection was achieved with stable nonunion (1 patient) and stable nonunion with cement spacer (2 patients). In 95% of the patients (19 of 20 patients) control of infection was achieved except for in 1 patient, who had a bony union with intermittent wound discharge and subsequently underwent an above-the-knee amputation. Three patients (15%) needed exchange nailing to another antibiotic cement-coated nail (for continued infection) before complete control of infection could be achieved. Four patients (20%) experienced cement-nail debonding during removal of the antibiotic cement-coated nail (3 during exchange to an uncoated intramedullary nail, 1 during removal at the request of patient). One patient experienced partial debonding at insertion, coinciding with the site of segmental defect, which was treated with an antibiotic cement spacer. In summary, control of infection and stability to promote union has traditionally been provided by 2 separate procedures, which have proved to be efficacious in the past. However, both these goals can be achieved in half the patients with 1 surgical procedure in a variety of scenarios using the technique of an antibiotic cement-coated intramedullary nail. PMID- 17414555 TI - Stabilization of unstable pelvic fractures with supraacetabular compression external fixation. AB - External fixation for pelvic stabilization is an important component of the overall treatment of patients with high-energy pelvic fractures. Traditional constructs include single and multiple pin placements in several locations in each iliac crest. Biomechanical and anatomic studies have focused on pin placement lower on the pelvis, specifically in the supraacetabular region. Pins in this location are more stable biomechanically, allow for pelvic reduction in the transverse plane of deformity, facilitate concurrent or subsequent laparotomy procedures, and may allow improved reduction of the posterior elements with a femoral distractor as a compressor. We describe the technique for placement of supraacetabular external fixation pins, pelvic reduction, and compression using a femoral distractor. PMID- 17414556 TI - Extended (16-hour) tourniquet application after combat wounds: a case report and review of the current literature. AB - We present a case of emergency tourniquet use of unusually long duration. The patient was wounded during combat operations, and the subsequent battle and evacuation caused a significant delay in surgical treatment of his wounds. Emergency tourniquets can be lifesaving, but are not benign interventions. In general, the extent of tourniquet injury increases with increasing time of application. Despite having a tourniquet in place for 16 hours, the limb was salvaged and significant functional recovery was accomplished. We conducted a search of the published literature including the Medline database, and present a review of the relevant articles concerning emergency tourniquet use, tourniquet injury, and mitigating treatments. Given the widespread use of tourniquets in ongoing military operations, it seems likely that tourniquets will transition to civilian use. Thus it is important for physicians to understand tourniquet injury and appreciate that even extended tourniquet application times does not necessarily doom the affected limb. PMID- 17414557 TI - Endovascular treatment of anterior tibial artery pseudoaneurysm following locking compression plating of the tibia. AB - Less invasive surgery and interventional radiology are relatively new techniques. This case report describes a patient with a distal tibial fracture that was stabilized using minimally invasive osteosynthesis consisting of a precontoured metaphyseal Locking Compression Plate (LCP). Postoperative radiographs showed good alignment of the bone, and the initial postoperative course was uneventful. At the sixth-week follow-up visit after surgery, the patient presented with a pulsating and tender mass on the lower leg that was palpable subcutaneously. Arteriography showed a pseudoaneurysm of the anterior tibial artery. At the same procedure an endovascular stent was placed, thereby excluding the pseudoaneurysm from the main circulation while keeping the vessel lumen patent. At the time of the last visit, 6 months after the operation, the patient was fully weightbearing with normal function of the ankle but with a nonhealing fracture on the x-ray. The dorsalis pedis pulse was equally strong as on the right side. Endovascular treatment with a covered stent proved to be an effective treatment for the described posttraumatic pseudoaneurysm of the anterior tibial artery. This case illustrates a risk of less invasive fracture surgery and at the same time underlines the value of a multidisciplinary approach to complications in trauma surgery. PMID- 17414559 TI - Complex limb salvage early amputation for severe lower-limb injury. PMID- 17414560 TI - Megadose methylprednisolone therapy: 2 decades of experience and empiric success in need of explanation, validation, and exploitation. PMID- 17414561 TI - A Tribute to Jon Pritchard, BA (Cantab), MB BChir (Cantab), MRCP (UK), FRCP (London), FRCPCH, FRCP (Edin): March 7, 1942 to January 20, 2007. PMID- 17414562 TI - Use of FEIBA for invasive or surgical procedures in patients with severe hemophilia A or B with inhibitors. AB - Achieving hemostasis in patients with hemophilia A or B is complicated by the presence of inhibitors and is made even more difficult when these individuals require surgery. Over a 4-year period, 6 patients with inhibitors to factor VIII and 1 patient with inhibitors to factor IX underwent surgery or invasive procedures at our institution. A total of 26 procedures were performed, primarily using the bypassing agent FEIBA for bleeding control. Excellent hemostasis was obtained in all cases, adding to accumulating data indicating that FEIBA is safe and effective in hemophilia patients with inhibitors who require surgery. PMID- 17414563 TI - Feasibility and toxicity of intrathecal liposomal cytarabine in 5 children and young adults with refractory neoplastic meningitis. AB - Intrathecal (IT) treatment of neoplastic meningitis secondary to relapsed or refractory malignancies is a major challenge for clinicians. We studied feasibility and toxicity of IT administered liposomal cytarabine on a compassionate basis in 5 patients (male, n=4; female, n=1; age at diagnosis 5 to 18 y) with recurrent acute lymphoblastic leukemia (n=3), primary refractory acute myeloid leukemia (n=1), or relapsed medulloblastoma (n=1). All of them had evidence of meningeal involvement as shown by presence of leukemic blasts or solid tumor cells on cytologic examination of cerebrospinal fluid and were refractory to standard central nervous system (CNS) therapy. A total of 33 doses were given. Leukemic blasts or solid tumor cells were cleared from cerebrospinal fluid in all patients. IT liposomal cytarabine was well tolerated in 2 patients, but may have contributed to neurologic side effects in 2 other patients with 1 patient who received high-dose methotrexate 96 hours before IT liposomal cytarabine developing transient encephalopathy. Another patient experienced seizures after 6 well-tolerated doses of IT liposomal cytarabine. In the fifth patient, treatment with IT liposomal cytarabine was not continued after a single dose because of toxic cauda equina syndrome, resulting from previous intensive CNS therapy. If administered simultaneously to other neurotoxic drugs, IT liposomal cytarabine may contribute to neurologic side effects in patients who had received prior intensive CNS-directed therapy. IT liposomal cytarabine should, therefore, be used cautiously, if a patient receives other potentially neurotoxic drugs simultaneously. PMID- 17414564 TI - Palivizumab prophylaxis to prevent respiratory syncytial virus mortality after pediatric bone marrow transplantation: a decision analysis model. AB - OBJECTIVE: Palivizumab, a monoclonal antibody against respiratory syncytial virus (RSV), has been demonstrated to be safe and effective in young children, but evidence is lacking as to whether palivizumab is effective in preventing RSV induced morbidity and mortality in children who are immunosuppressed after bone marrow transplantation (BMT). As a randomized, double-blind, placebo-controlled trial is lacking, we chose to examine this issue with the use of decision analysis methodology. METHODS: A decision tree was designed to determine mortality from RSV-related lung disease in children who received palivizumab after BMT. Probabilities were derived by meta-analysis methodology on the basis of the available literature. Sensitivity analyses were performed across a broad range of biologically plausible probabilities to judge the robustness of the results of the model. RESULTS: The model revealed that there is a 10% increase in survival in BMT patients who receive palivizumab. The absolute survival rate increased from 83% to 92%. A practitioner would need to treat 12 children to save 1 post-BMT child from dying from RSV-related lung disease. CONCLUSIONS: Decision analysis modeling demonstrates a decrease in mortality in pediatric BMT patients with the addition of palivizumab to protect against RSV-related lung disease. A well-designed, randomized controlled trial is necessary. PMID- 17414565 TI - Thalassemia in Iran: epidemiology, prevention, and management. AB - PURPOSE: To determine the prevalence and geographic distribution of thalassemia and to evaluate the success of the thalassemia prevention and treatment programs in Iran. METHODS: Data were obtained from the National Thalassemia Registry of Iran, Iranian Blood Transfusion Organization, genetic laboratories involved in prenatal diagnosis, related pharmaceutical companies, and centers performing bone marrow transplantation for thalassemic patients. RESULTS: A total of 13,879 living patients have been registered, mostly from the northern and southern parts of Iran with the median age of 15 years. Twenty-three percent of patients were older than 20 years. The number of newly diagnosed cases has been decreased considerably after the start of the prevention program. Since the introduction of prenatal diagnosis, 2819 couples (2549 fetuses) have been tested, with only 6 false results. Elective abortion was not performed in 10 affected fetuses. Most common mutations detected were IVS II-1 and IVS I-5. In 2003, approximately 25% of the national blood products and 6 million vials of desferal were used for thalassemic patients. Overall, 340 patients have received allogeneic bone marrow transplantation, of those 46 patients deceased. Bloodborne infections have also been decreased significantly owing to the national screening of blood products for bloodborne viral infections. DISCUSSION: Owing to the national prevention program and provided special care, the age distribution of thalassemic patients in Iran is getting adapted to a full prevention and treatment program and life expectancy of these patients has been increased considerably. This shift in the age distribution of thalassemia, a traditionally considered pediatric disease, will face us with new challenges and the health care system should be prepared for this new face of thalassemia. PMID- 17414566 TI - Clinical pharmacology of asparaginases in the United States: asparaginase population pharmacokinetic and pharmacodynamic (PK-PD) models (NONMEM) in adult and pediatric ALL patients. AB - In the past 25 years, effective new drugs along with better treatment decisions based on disease factors have resulted in significantly improved clinical outcomes in acute lymphoblastic leukemia. Despite these successes in the last 2 decades, 15% to 25% of acute lymphoblastic leukemia patients relapse. Therefore, better dosing therapies are still needed. Insights in the pharmacokinetic and pharmacodynamic (PK-PD) contributions of licensed drugs may guide us into better protocol design and optimal use of existing combination drug regimens. Currently, 3 asparaginase formulations are available in the United States, Escherichia coli native asparaginase (ASNase), Pegaspargase, and Erwinase. On the basis of these clinical studies, PK and PD population modeling (NONMEM) have been used to delve into new insights as to the optimal dose, formulation, and time intervals of ASNases that may be used in future clinical trials. Pegaspargase 2500 IU/m2 Q2week dosing seems to be the "golden standard" as far as being safe and effective. Lower doses of this formulation Qweek may achieve better PK "steady state" profiles in serum. Native E. coli or Erwinia ASNase at 6000 IU/m2 showed inferior PK parameters (peak, trough, and area under the curve) than Pegaspargase. Assuming linear handling of ASNase modeling, simulations of higher doses of these ASNase formulations on a daily or Q48 hours regimen are showing bioequivalency with Pegaspargase PK-PD parameters. Future clinical trial designs may prove these efforts useful. PMID- 17414567 TI - Secondary glioblastoma multiforme after treatment for primary choroid plexus carcinoma in childhood. AB - A 15-year-old boy was diagnosed with choroid plexus carcinoma (CPC) of the right lateral ventricle. His metastatic work-up was negative. After complete macroscopic resection of the tumor, the patient was treated with chemotherapy consisting of vincristine, cisplatin, etoposide, and carboplatin, followed by radiotherapy for a total dose of 34.2 Gy on the whole craniospinal axis plus a boost of 19.8 Gy at the tumor region. The patient remained in complete clinical and radiologic remission over the next 5 years when a secondary malignant tumor, glioblastoma multiforme, a rare complication of the treatment of CPC, was diagnosed. This case reflects the necessity of thorough follow-up in long-term survivors of CPC. PMID- 17414568 TI - Comparison of megadose methylprednisolone versus conventional dose prednisolone in hematologic disorders. AB - Glucocorticoids (GCs) are known for their clinically useful effects in immunologic and inflammatory disorders. Although there is a huge volume of knowledge concerning the cellular and molecular effects of GCs, statements regarding their effects in multiple diseases at variable doses are not clear-cut owing to pharmacogenetic differences. The main actions of GCs in hematologic disorders have been related to their differentiation-inducing and apoptosis inducing effects, but modification of several steps of the hematopoietic and/or immune pathway has also been reported. In our clinic, mega-dose methylprednisolone (MDMP) has been successfully used for treatment of different hematologic diseases, such as leukemias, bone marrow failure in aplastic anemia, hypoplastic anemia, myelodysplastic syndrome, neutropenia, autoimmune diseases, and in some congenital hereditary diseases. Both clinical and experimental studies in our department revealed that MDMP was more effective than conventional dose steroids. It is interesting that MDMP can be curative in some congenital hereditary diseases such as Diamond-Blackfan syndrome. However, more research is required to clarify their roles in biology, physiology, and molecular genetics. PMID- 17414569 TI - Neuroblastoma masquerading as cervical lymphadenitis. AB - Cervical lymphadenitis is a common pediatric finding prompting medical evaluation. The most common etiologies are infectious and reactive. The location of the involved lymph node group may provide clues to the origin of the underlying pathologic process. We describe a 21-month-old boy with metastatic neuroblastoma who presented with classic findings of infectious lymphadenitis. Surgical intervention and careful examination of histopathology led to this unexpected diagnosis. PMID- 17414570 TI - Respiratory failure, juvenile myelomonocytic leukemia, and neonatal Noonan syndrome. AB - Noonan syndrome (NS) is a multiple malformation syndrome where confirmation of diagnosis is difficult in the newborn. We report a case of a dysmorphic neonate who presented with bilateral chylous effusions and juvenile myelomonocytic leukemia where NS was confirmed by the presence of PTPN11 mutation. Juvenile myelomonocytic leukemia in NS is uncommon. The leukemia is usually self-limiting but lethal cases have been reported. Decisions regarding need for the treatment are unclear and further understanding of the genotype-phenotype relationships in PTPN11 mutations may help direct this. PMID- 17414571 TI - Argatroban for anticoagulation in pediatric patients with heparin-induced thrombocytopenia requiring extracorporeal life support. AB - Heparin-induced thrombocytopenia (HIT) is a well recognized and described immune phenomenon in adults. However, pediatric data on the incidence, treatment, and complications of HIT are limited. Despite the widespread use of heparin in pediatric patients and the repeated and sustained exposures to heparin in neonates and children with congenital heart disease, HIT has been infrequently recognized and under-reported in this population. HIT is an intensely procoagulant disorder and carries significant morbidity and mortality. In this case series, we describe HIT management with argatroban as alternative anticoagulation in 3 children requiring extracorporeal life support. PMID- 17414572 TI - Assessing the outcome of systemic tissue plasminogen activator for the management of venous and arterial thrombosis in pediatrics. AB - This study sought to ascertain the outcomes of systemic thrombolytic therapy used in a cohort of infants and children. Complete thrombus resolution was achieved in 81% of patients with arterial thromboses (n=16) compared to 0% of children with venous thromboses (n=10). A major bleeding rate of 11.5% occurred across the entire cohort (n=3, all arterial). In our cohort, no patient with venous thromboembolism achieved complete resolution of their thrombosis after thrombolytic therapy. More cohort studies reporting the outcome of uniform protocols of thrombolytic therapy in children are required. PMID- 17414573 TI - A case of near-triploid with t(2;14)(p12;q32) in blast crisis of chronic myeloid leukemia detected by spectral karyotyping (SKY). PMID- 17414574 TI - "Flushing out" cytosine arabinoside from CSF to reverse neurotoxicity. PMID- 17414576 TI - Drospirenone, a progestin with added value for hypertensive postmenopausal women. PMID- 17414577 TI - The drug safety review process. PMID- 17414579 TI - The future of antihypertensive treatment. AB - Despite progress in recent years in the prevention, detection, and treatment of high blood pressure (BP), hypertension remains an important public health challenge. Hypertension affects approximately 1 billion individuals worldwide. High BP is associated with an increased risk of mortality and morbidity from stroke, coronary heart disease, congestive heart failure, and end-stage renal disease; it also has a negative impact on the quality of life. Hypertension cannot be eliminated because there are no vaccines to prevent the development of hypertension, but, its incidence can be decreased by reducing the risk factors for its development, which include obesity, high dietary intake of fat and sodium and low intake of potassium, physical inactivity, smoking, and excessive alcohol intake. For established hypertension, efforts are to be directed to control BP by lifestyle modification (LSM). However, if BP cannot be adequately controlled with LSM, then pharmacotherapy can be instituted along with LSM. Normalization of BP reduces cardiovascular risk (for cardiovascular death, myocardial infarction, and cardiac arrest), provides renoprotection (prevention of the onset or slowing of proteinuria and progression of renal dysfunction to end-stage renal disease in patients with hypertension, diabetes mellitus types 1 and 2, and chronic renal disease), and decreases the risk of cerebrovascular events (stroke and cognition impairment), as has been amply demonstrated by a large number of randomized clinical trials. In spite of the availability of more than 75 antihypertensive agents in 9 classes, BP control in the general population is at best inadequate. Therefore, antihypertensive therapy in the future or near future should be directed toward improving BP control in treated hypertensive patients with the available drugs by using the right combinations at optimum doses, individually tailored gene-polymorphism directed therapy, or development of new modalities such as gene therapy and vaccines. Several studies have shown that BP can be reduced by lifestyle/behavior modification. Although, the reductions appear to be trivial, even small reductions in systolic BP (for example, 3-5 mm Hg) produce dramatic reduction in adverse cardiac events and stroke. On the basis of the results of clinical and clinical/observational studies, it has been recommended that more emphasis be placed on lifestyle/behavior modification (obesity, high dietary intake of fat and sodium, physical inactivity, smoking, excessive alcohol intake, low dietary potassium intake) to control BP and also to improve the efficacy of pharmacologic treatment of high BP. New classes of antihypertensive drugs and new compounds in the established drug classes are likely to widen the armamentarium available to combat hypertension. These include the aldosterone receptor blockers, vasodilator beta-blockers, renin inhibitors, endothelin receptor antagonists, and dual endopeptidase inhibitors. The use of fixed-dose combination drug therapy is likely to increase. There is a conceptual possibility that gene therapy may yield long-lasting antihypertensive effects by influencing the genes associated with hypertension. But, the treatment of human essential hypertension requires sustained over-expression of genes. Some of the challenging tasks for successful gene therapy that need to be mastered include identification of target genes, ideal gene transfer vector, precise delivery of genes into the required site (target), efficient transfer of genes into the cells of the target, and prompt assessment of gene expression over time. Targeting the RAS by antisense gene therapy appears to be a viable strategy for the long-term control of hypertension. Several problems that are encountered in the delivery of gene therapy include 1) low efficiency for gene transfer into vascular cells; 2) a lack of selectivity; 3) problem in determining how to prolong and control transgene expression or antisense inhibition; and 4) difficulty in minimizing the adverse effects of viral or nonviral vectors. In spite of the hurdles that face gene therapy administration in humans, studies in animals indicate that gene therapy may be feasible in treating human hypertension, albeit not in the near future. DNA testing for genetic polymorphism and determining the genotype of a patient may predict response to a certain class of antihypertensive agent and thus optimize therapy in individual patients. In this regard, there are some studies that report the effectiveness of antihypertensive therapy based upon the genotype of selected patients. Treatment of human hypertension with vaccines is feasible but is not likely to be available in the near future. PMID- 17414580 TI - Management of hypertensive crises. AB - Hypertensive emergencies are life-threatening conditions because their course is complicated with acute target organ damage. They can present with neurological, renal, cardiovascular, microangiopathic hemolytic anemia, and obstetric complications. After diagnosis, they require the immediate reduction of blood pressure (in <1 hour) with intravenous drugs such as sodium nitroprusside, administered in an intensive care unit. These patients present with a mean arterial pressure >140 mm Hg and grade III to IV retinopathy. Only occasionally do they have hypertensive encephalopathy, reflecting cerebral hyperperfusion, loss of autoregulation, and disruption of the blood-brain barrier. In hypertensive emergencies, blood pressure should be reduced about 10% during the first hour and another 15% gradually over the next 2 to 3 hours to prevent cerebral hypoperfusion. The exception to this management strategy is aortic dissection, for which the target is systolic blood pressure <120 mm Hg after 20 minutes. Oral antihypertensive therapy can usually be instituted after 6 to 12 hours of parenteral therapy. Hypertensive urgencies are severe elevations of blood pressure without evidence of acute and progressive dysfunction of target organs. They demand adequate control of blood pressure within 24 hours to several days with use of orally administered agents. The purpose of this review is to provide a rational approach to hypertensive crisis management. PMID- 17414581 TI - A double-blind, controlled, multicenter, randomized study comparing the antihypertensive effectiveness and tolerance of a daily dose of two nifedipine formulations: nifedipine microgranules versus nifedipine osmotic pump. AB - INTRODUCTION: Controlled clinical studies have clearly established the advantages of blood pressure (BP) reduction. However, optimal control of BP in the population is still not adequate. Monotherapy is ineffective in the majority of hypertensive patients, and multidrug therapy increases costs. OBJECTIVE: The objective of the study was to assess to what extent and how uniformly BP can be controlled with two different 24-hour drug-releasing formulations of nifedipine, used as monotherapy. METHODS: One hundred ninety-two patients of both genders, aged 18 to 65 years, with mild to moderate (Stage 1 and 2) essential hypertension with systolic BP <200 mm Hg and diastolic BP between 90 and 115 mm Hg were randomized in a double-blind, double-dummy fashion to receive sustained-release formulations of 30 mg nifedipine/day either as microgranules (NMG) or via osmotic pump (NOP) for 8 weeks. Office BP was measured at baseline (after 2 weeks of placebo) and after the third to fourth week of treatment. If at the third to fourth week the systolic BP/diastolic BP did not reach values of <140/<90 mm Hg, the dose was doubled to 60 mg/day. Monotherapy that did not yield these BP values at 8 weeks was considered a failure. Ambulatory monitoring of blood pressure (AMBP) was also performed after the placebo period and at the end of treatment. Smoothness index (SI) and trough/peak ratio (T/P) were calculated and their correlation was checked. RESULTS: The initial systolic/diastolic BP values were similar at baseline and decreased significantly after the third to fourth week of treatment, with no difference between the groups. The proportions of patients reaching the goal BP (<140/<90 mm Hg) were similar in the two groups: NMG, 71%, and NOP, 78% (P = 0.12). There were no changes in the heart rate in either group. There was no difference between groups in the reduction in mean arterial pressure measured by AMBP. The frequency of SI values >1.4 and T/P ratios of >0.5 was similar in both groups. An important correlation was found between the SI and T/P values. The incidence of adverse effects was low and similar in both groups. CONCLUSIONS: Target BP was reached in more than 70% of patients receiving monotherapy with either formulation. Both formulations were tolerated well. PMID- 17414582 TI - Role of endothelins in hypertension. AB - There are three peptides of endothelial origin, called endothelins (ETs), having different receptors that mediate a potent vasoconstrictor effect and also a mild vasodilation. Their renal effects are characterized by natriuresis in spite of the renal vasoconstriction. This effect, along with the stimulation of ETs by high sodium intake, suggests that ETs may be responsible for maintaining sodium balance when the renin angiotensin system is depressed. ET is activated in deoxycorticosterone acetate (DOCA) salt hypertension models and salt-sensitive hypertension. In humans, the role of ET seems to be similar to that shown in experimental animals; in both, ET participates in the regulation of salt metabolism. Salt-sensitive patients exhibit a blunted renal ET-1 response during sodium load. The role of ETs in humans has been investigated with use of nonspecific ET receptor blockers that inhibit the vasoconstriction and vasodilator components of ET. However, the effects of ET blockade should be investigated with ETA receptor blockers that mediate vasoconstriction alone. Effects of ET blockade should also be evaluated with respect to stimulation of oxidative stress and tissue damage, important mechanisms responsible for tissue fibrosis. PMID- 17414583 TI - Update of diuretics in the treatment of hypertension. AB - Diuretics, which are primarily used to modify the volume and the composition of body fluids, are widely used to treat hypertension. The diuretics include a) the thiazides and thiazide-like agents, which are the most common drugs used to treat high blood pressure (these drugs inhibit sodium reabsorption in the early distal convoluted tubule); b) loop diuretics, such as furosemide, block chloride and sodium reabsorption by inhibition of the Na/K/2Cl cotransport system in the thick ascending limb of the loop of Henle; and c) potassium-sparing (retaining) diuretics, including aldosterone receptor blockers (such as spironolactone and eplerenone) and epithelial sodium channel blockers (such as amiloride and triamterene, which interfere with the reabsorption of sodium and excretion of potassium and hydrogen that takes place in the late distal tubule, the connecting tubule, and the cortical collecting duct). Hydrochlorothiazide 12.5 mg once daily or equivalent low dosages of other similar agents reduce blood pressure in approximately one-half to two-thirds of patients who are responsive to this class of drugs; higher doses add little to the effect on blood pressure and also increase side effects. Some combinations of very small doses of thiazide diuretics - for example, 6.25 mg hydrochlorothiazide or 0.625 mg indapamide, with a low dose of an antihypertensive drug of a different class - have average antihypertensive efficacy when used once daily. Furosemide is used in patients with renal failure or severe heart failure and is best given by continuous intravenous infusion. The potassium-sparing diuretics are generally used in combination with thiazide diuretics to treat hypertension. Side effects occur at about the same frequency and severity with equipotent doses of all diuretics. The incidence of side effects is dose-dependent and also increases as a function of the duration of the renal excretory and antihypertensive actions. However, longer acting diuretics provide better 24-hour control of blood pressure and increase compliance and adherence to the treatment regimen. PMID- 17414584 TI - Effect of losartan plus hydrochlorothiazide on nitric oxide status in 'nondipper' hypertensive patients. AB - The objective of this study was to investigate the effects of losartan (100 mg) plus hydrochlorothiazide (HCTZ; 25 mg) on nitric oxide (NO) production and blood pressure (BP) in "nondipper" severe hypertensive patients. Twelve hypertensive "nondipper patients" (6 of each gender) with sitting systolic/diastolic BP of 188.0 +/- 5.2/116.2 +/- 1.2 mm Hg were studied by 24-hour ambulatory blood pressure monitoring (ABPM) after daily administration of 100 mg losartan plus 25 mg HCTZ for a period of 12 weeks. Office and mean 24-hour, as well as mean awake- and sleep-time systolic/diastolic BP, serum NO levels, and urinary excretion of NO were measured after the placebo period (3 weeks) and after 12 weeks of therapy. At the end of the 12-week treatment period, the mean 24-hour systolic/diastolic BP decreased significantly from 158.6 +/- 4.7/102.2 +/- 2.6 mm Hg (placebo period) to 140.3 +/- 4.8/90.9 +/- 3.3 mm Hg (P = 0.001/< or = 0.002). The mean BP (systolic/diastolic) during the waking period was reduced from 159.3 +/- 4.4/103.0 +/- 2.5 mm Hg to 135.0 +/- 4.4/88.2 +/- 3.1 (P < or = 0.007/P < or = 0.002), whereas the mean BP (systolic/diastolic) during the sleeping hours changed from 154.9 +/- 5.3/98.9 +/- 3.1 to 140.9 +/- 4.6 (P = 0.035)/91.7 +/- 3.2 mm Hg (P = 0.035/P = 0.051). Serum NO levels increased from 40.89 +/- 5.69 microM/L (placebo period) to 67.35 +/- 6.96 microM/L (posttreatment; P < or = 0.007), whereas the 24-hour urinary NO excretion did not change significantly (69.71 +/- 3.68 microM/L [placebo period] vs 79.64 +/- 4.25 microM/L [posttreatment]; P < or = 0.16). Urinary clearance of NO also did not change. Serum NO levels increased significantly without a significant change in urinary NO excretion. BP was significantly reduced but without modifying the nondipper pattern in these patients. PMID- 17414585 TI - Effect of losartan therapy on endothelial function in hypertensive patients. AB - The aim of the study was to evaluate the effect of losartan therapy on endothelial function by measuring serum nitric oxide (NO) levels and urinary excretion of NO in patients with essential hypertension. A group of 30 untreated stage 2 hypertensive patients (15 males and 15 females; age, 51.3 +/- 1.5 years) were included in the study. Office systolic and diastolic blood pressure (BP) was measured by using a mercury sphygmomanometer according to phase I and V of Korotkoff sounds. NO levels in serum and 24-hour urine were determined at baseline and after 6 weeks of daily dosing with losartan (50-100 mg). Losartan therapy resulted in a significant fall in systolic/diastolic BP (from 169.7 +/- 4.1/105 +/- 1.8 mm Hg at baseline to 146 +/- 2.7/91 +/- 1.9 mm Hg at the end of losartan treatment; P < 0.001). The therapy also caused significant increases in both serum NO level (32.74 +/- 3.01 microM/L at baseline versus 79.04 +/- 5.17 microM/L; P < 0.001 after therapy) and urinary NO excretion (58.21 +/- 3.72 microM/L at baseline versus 113.21 +/- 8.63 microM/L; P < 0.001 after therapy). Losartan therapy also reduced serum malondialdehyde (MDA), which is a measure of oxidative stress, by 0.201 nM (15.3%; P = 0.009). Losartan at a dose of 50 to 100 mg per day was effective in reducing elevated BP. The increase in serum NO levels and urinary NO excretion and a decrease in serum MDA levels by losartan treatment indicate a reduction in oxidative stress and enhances NO availability, both of which improve endothelial function. Thus, losartan therapy improves endothelial function in hypertensive patients with essential hypertension. PMID- 17414586 TI - Nitric oxide and malondialdehyde in human hypertension. AB - Nitric oxide (NO), a multifunctional effector molecule that plays a central role in the maintenance of vascular homeostasis, regulates vascular tone and inhibits platelet and leukocyte adhesion to endothelial cells. NO status is related to the endothelial function. Patients with hypertension have lower levels of NO, increased free radical production, higher oxidative stress, augmented platelet aggregation, and a change in the arachidonic acid cascade metabolism, all leading to the acceleration of the atherosclerotic process. The study subjects included a group of 21 normotensive healthy subjects (8 males and 13 females) with a mean age of 39.2 +/- 1.8 years and a body mass index of 27.9 kg/m, and another group of 42 patients (19 males and 23 females) with untreated essential hypertension with a mean age of 47.6 +/- 1.7 years and a body mass index of 28.3 kg/m. Serum levels and urinary excretion of NO determined as combined nitrate/nitrite (NOx) and serum malondialdehyde (MDA) concentrations were measured in the 2 groups of subjects. The serum levels and 24-hour urinary excretion of NOx were significantly higher and the renal clearance of NO was lower in the normotensive group than in the hypertensive patients, indicating decreased NO status in hypertension. There was a negative correlation between serum NO levels and mean arterial pressure, suggesting that a decrease in NO availability is related to increase in blood pressure. Serum concentrations of MDA were higher in the hypertensive patients as compared with the normotensive individuals, suggesting increased oxidative stress in hypertensive patients. These results are in agreement with previous studies showing decreased NO and increased oxidative stress in hypertension. In conclusion, patients with essential hypertension as compared with normotensive individuals have lower NO status, which may contribute to the endothelial dysfunction in hypertension. Increased serum malondialdehyde in hypertensives suggests an association between increased oxidative stress with higher blood pressure. PMID- 17414587 TI - Fibrinolytic system in normotensive subjects and hypertensive patients. AB - The aim of this study was to evaluate the fibrinolytic system by measurement of fibrinogen, plasminogen, tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor-1 (PAI-1) in healthy normotensive subjects and in patients with essential hypertension. A group of 21 healthy normotensive subjects [age, 39.2 +/- 1.8 years; 8 males, 13 females; body mass index (BMI) = 27.9 kg/m] and 42 patients with untreated essential hypertension (age, 47.6 +/- 1.7 years; 19 males, 23 females; BMI = 28.3 kg/m) were studied. Blood samples and clinical measurement were taken between 7 am and 9 am by an observer in a blind fashion. The systolic/diastolic blood pressure of normotensive subjects was 121.3 +/- 2.5/78.4 +/- 1.3 mm Hg and that of hypertensive patients was 166.4 +/- 4.3/102.9 +/- 1.83 mm Hg, measured in the sitting position. Plasma fibrinogen levels in the normotensive and hypertensive individuals were 295.7 +/- 9.4 mg/dL and 305.67 +/- 10.9 mg/dL, respectively (P = 0.456). The corresponding values for plasminogen were 71.4 +/- 3.8% and 89.5 +/- 2.5%, (P = 0.0031), for t-PA were 6.3 +/- 0.5 ng/mL and 7.6 +/- 0.4 ng/mL (P = 0.0487), and for PAI-1 were 46.9 +/- 5.1 ng/mL and 63.0 +/- 5.6 ng/mL (P = 0.0324), respectively. In conclusion, patients with essential hypertension have disequilibrium in the fibrinolytic system with a tendency toward a hypercoagulability state when compared with normotensive subjects. This state could explain, in part, the thrombotic complications that occur with a higher frequency in hypertensive patients as compared with normotensive subjects. PMID- 17414588 TI - Effect of AT(1) angiotensin II receptor antagonists on the sympathetic response to a cold pressor test in healthy volunteers. AB - The aim of this study was to evaluate the effect of short-term administration of AT(1) angiotensin II receptor antagonists on the sympathetic response to a cold pressor test (CPT) in normotensive healthy volunteers. Eighty-two healthy volunteers were included in this double-blind placebo-controlled study. Blood pressure and heart rate were determined before and 175 minutes after oral administration of placebo, losartan (50 mg), valsartan (80 mg), or eprosartan (600 mg). Immediately, the subjects underwent a CPT and then the same hemodynamic parameters were measured. CPT increased arterial blood pressure (systolic, diastolic, and mean) and heart rate in the placebo-treated group. Pretreatment with a single dose of losartan, valsartan, or eprosartan blunted CPT-induced pressor response but not heart rate increase. Our results demonstrate that endogenous angiotensin II, through stimulation of AT(1) receptor, supports sympathetic-mediated stress response in humans. PMID- 17414589 TI - Effect of cigarette smoking on the oxidant/antioxidant balance in healthy subjects. AB - BACKGROUND AND PURPOSE: Cigarette smoking has been associated with the development of cardiovascular disease and cancer. Even though the molecular mechanism(s) are not clear, the pathology has been related to oxygen free radicals present in cigarette smoke. Thus, the main objective of this study was to establish the changes in the oxidation/antioxidation balance induced by cigarette smoking. METHODS: Thirty healthy subjects (15 smokers and 15 nonsmokers) of both sexes were studied. The smokers group had smoked a mean of 14 cigarettes per day for an average of 4.5 years. Fasting serum levels of malondialdehyde (MDA), a marker of oxidative stress, nitric oxide (NO), reduced glutathione (GSH), and vitamin C (ascorbic and dehydroascorbic acids) were measured. RESULTS: Fasting NO concentration was significantly higher in smokers (51.3 +/- 5.3 microM) than in nonsmokers (35.2 +/- 4.8 microM, P < 0.05). The smokers had significantly higher serum dehydroascorbic acid levels (2.4 +/- 0.5 mg/dL, P < 0.03) than the nonsmokers (1.08 +/- 0.08 mg/dL). No significant differences were observed in the levels of ascorbic acid, MDA, and GSH between the smokers and nonsmokers. CONCLUSIONS: Our results suggest that exposure to cigarette smoke increases NO synthesis, such that NO may act in a compensatory way as an inhibitor of lipid peroxidation. Smoking also activates other antioxidative mechanisms such as involving vitamin C. These protective mechanisms appear to be enough in preventing accumulation of oxidative products such as MDA and avoiding oxidative damage. PMID- 17414590 TI - Metformin plus low-dose glimeperide significantly improves Homeostasis Model Assessment for insulin resistance (HOMA(IR)) and beta-cell function (HOMA(beta cell)) without hyperinsulinemia in patients with type 2 diabetes mellitus. AB - OBJECTIVE: Type 2 diabetes mellitus is characterized by insulin resistance and defects in insulin secretion from pancreatic beta-cells, which have been studied by using euglycemic/hyperinsulinemic clamps. However, it is difficult to study insulin resistance and beta-cell failure by these techniques in humans. Therefore, the aim of this study was to evaluate the effect of three different antidiabetic therapeutic regimens on insulin resistance and beta-cell activity by using a mathematical model, Homeostasis Model Assessment for insulin resistance (HOMA(IR)) and beta-cell function (HOMA(beta-cell)). RESEARCH DESIGN AND METHODS: Seventy type 2 diabetic patients were randomly assigned to one of three therapeutic regimens: (A) metformin + American Diabetic Association (ADA) recommended diet + physical activity; (B) metformin + low-dose glimepiride + ADA diet + physical activity; or (C) ADA diet + physical activity (no drugs). Blood samples were obtained before and after the treatment to determine serum levels of fasting and post-prandial blood glucose, fasting insulin, and glycosylated hemoglobin (HbA1c), and HOMA(IR) and HOMA(beta-cell) were calculated. RESULTS: Fasting and post-prandial levels of glucose, HbA1c, and fasting insulin and calculated HOMA(IR) and HOMA(beta-cell) values before treatment were significantly higher than the respective values after treatment for all groups of patients (P < 0.01). Significant differences were also found when comparing the treatment-induced reduction in fasting blood glucose (51.8%; P < 0.01), post prandial blood glucose (55.0%; P < 0.05), and HOMA(IR) (65.3%; P < 0.01) in patients of Group B with that in patients receiving other therapeutic options (Groups A and C). CONCLUSIONS: Metformin plus low-dose glimepiride (plus ADA diet and physical activity) is a more effective treatment for type 2 diabetes than either metformin plus ADA diet and physical activity or ADA diet and physical activity alone. Determination of HOMA(IR) and HOMA(beta-cell) values is an inexpensive, reliable, less invasive, and less labor-intensive method than other tests to estimate insulin resistance and beta-cell function in patients with type 2 diabetes mellitus. PMID- 17414591 TI - Oat-derived beta-glucan significantly improves HDLC and diminishes LDLC and non HDL cholesterol in overweight individuals with mild hypercholesterolemia. AB - OBJECTIVE: To investigate the effect of bread formulated with 6 g of beta-glucan (oat soluble fiber) on serum lipids in overweight normotensive subjects with mild to moderate hypercholesterolemia. DESIGN: Thirty-eight male subjects [mean age 59.8 +/- 0.6 yr, mean body mass index (BMI) 28.3 +/- 0.6 kg/m(2)] who were eligible for the study ate an isocaloric diet for a 1-week period. They were then divided into 2 groups: group A (n = 19), who were maintained on American Heart Association (AHA) Step II diet, including whole wheat bread, and group B (n = 19), who were maintained on AHA Step II diet containing high levels of monounsaturated fatty acids plus bread containing 6 g of beta-glucan (Nutrim-OB) for 8 weeks. Plasma lipids and glucose were measured at baseline and after weeks 8 in all subjects. All subjects were advised to walk for 60 minutes every day. RESULTS: There was a significant increase (upward arrow 27.8%) in plasma high density lipoprotein (HDL) cholesterol in the beta-glucan group (group A) from 39.4 +/- 2.0 to 49.5 +/- 2.1 mg/dL (P < 0.001), but there was no change in group B. There was a significant reduction in total cholesterol in the 2 groups to approximately the same extent: group A, from 232.8 +/- 2.7 mg/dL to 202.7 +/- 6.7 mg/dL; P < 0.001; and group B, from 231.8 +/- 4.3 mg/dL to 194.2 +/- 4.3 mg dL; P < 0.001. Plasma low density lipoprotein (LDL) cholesterol also decreased significantly in the two groups: group A, from 160.3 +/- 2.8 mg/dL to 133.2 +/- 5.4 mg/dL; P < 0.001; group B, from 167.9 +/- 4.3 mg/dL to 120.9 +/- 4.3 mg/dL; P < 0.001; however, the beta-glucan fortified diet was significantly more effective (downward arrow 27.3% vs. downward arrow 16.8%; P < 0.04). There was a small and insignificant reduction in plasma very LDL (VLDL) cholesterol and triglycerides in the two groups. Similarly, non-HDL cholesterol levels were also decreased, with beta-glucan diet producing significantly higher effect (downward arrow 24.5% vs. downward arrow 16.1%; P < 0.04). The beta-glucan diet also produced higher reduction in total cholesterol/HDL cholesterol ratio (downward arrow 33.3% vs. downward arrow 8.4%; P < 0.003) and LDL cholesterol/HDL cholesterol ratio (downward arrow 42.1% vs. downward arrow 13.3%; P < 0.001) than the diet without beta-glucan. The beta-glucan diet also decreased fasting plasma glucose (P < 0.4), whereas the other diet had no effect. Interestingly, both diets reduced body weight and BMI significantly, with beta-glucan diet having a greater effect. CONCLUSIONS: Six grams of beta-glucan from oats added to the AHA Step II diet and moderate physical activity improved lipid profile and caused a decrease in weight and, thus, reduced the risk of cardiovascular events in overweight male individuals with mild to moderate hypercholesterolemia. The diet with added beta glucan was well accepted and tolerated. PMID- 17414592 TI - Ciprofibrate treatment decreases non-high density lipoprotein cholesterol and triglycerides and increases high density lipoprotein cholesterol in patients with Frederickson type IV dyslipidemia phenotype. AB - OBJECTIVE: The combination of hypertriglyceridemia and low high density lipoprotein (HDL) cholesterol is one of the most common lipid abnormalities. Thus, the aim of this study was to determine the effects of ciprofibrate on lipid profile in patients with Frederickson's type IV dyslipidemia phenotype. RESEARCH DESIGN AND METHODS: Seventy-five patients with type IV dyslipidemia were assigned at random to 1 of 2 therapeutic options: group A (control), American Heart Association (AHA) Step II diet and physical activity; and group B, AHA diet, physical activity, and ciprofibrate 100 mg daily for 8 weeks. The lipid profile of all patients was determined at baseline and after therapeutic intervention. RESULTS: Patients in group B (treated with ciprofibrate) compared with group A (control) had significantly higher reductions in total cholesterol (downward arrow 14.2% vs. downward arrow 4.8%; P < 0.02), triglycerides (downward arrow 38.0% vs. downward arrow 21.6%; P < 0.007), very low density lipoprotein cholesterol (downward arrow 38.0% vs. downward arrow 21.6%; P < 0.007), non-HDL cholesterol (downward arrow 20.5% vs. downward arrow 7.1%; P < 0.007), and total cholesterol/high density cholesterol ratio (downward arrow 25.6% vs. downward arrow 9.4%; P < 0.01). The ciprofibrate group had a significantly higher increase in HDL cholesterol levels compared with the other group (upward arrow 25.0% vs. upward arrow 9.6%, P < 0.02). CONCLUSIONS: Ciprofibrate treatment effectively reduced triglyceride-rich particles and non-HDL cholesterol, and significantly increased HDL cholesterol, proving its effectiveness in patients with low HDL cholesterol and type IV Frederickson's hyperlipidemia. PMID- 17414593 TI - Once-daily bupropion associated with improved patient adherence compared with twice-daily bupropion in treatment of depression. AB - The main aim of this study was to examine the impact of dosing regimens on patients' persistence to bupropion. A nationally representative patient-level database comprising of pharmacy and medical claims was used to identify patients with depression (ICD-9-CM: 296.2, 296.3, 300.4, 311), who had initiated therapy with bupropion sustained release (b.i.d.; 2 doses/d) or extended release (q.d.; 1 dose/d) tablets from September 2003 to February 2004; had no previous antidepressant or benzodiazepine use; and had 9 months of follow-up. Persistence was measured using prescription claims, and calculated using the medication possession ratio [MPR; (total days supply; all filled prescriptions)/270 d]. Multivariate logistic regression compared the likelihood of achieving MPR > or = 0.70 controlling for age, sex, and index date. A total of 3132 patients were included (b.i.d.: n = 2382; q.d.: n = 756). q.d. patients on average had a significantly higher MPR than b.i.d. patients [q.d. 0.52 (+/-0.35), b.i.d. 0.35 (+/-0.26)]; P < 0.001) and had a higher likelihood of achieving an MPR > or = 0.70 (q.d. 35%, b.i.d. 12%, P < 0.0001). After controlling for differences in baseline characteristics, b.i.d. patients were only one-fourth as likely (odds ratio = 0.260, 95% confidence interval: 0.214-0.316) to achieve MPR >0.7. The use of the once-daily, extended release formulation of bupropion appeared to significantly improve patients' persistence to therapy for the treatment of depression. PMID- 17414594 TI - Consensus statement on mandatory registration of clinical trials. PMID- 17414595 TI - Disclosure of individual surgeon's performance rates during informed consent: ethical and epistemological considerations. AB - OBJECTIVE: The purpose of the paper is to examine the ethical arguments for and against disclosing surgeon-specific performance rates to patients during informed consent, and to examine the challenges that generating and using performance rates entail. METHODS: Ethical, legal, and statistical theory is explored to approach the question of whether, when, and how surgeons should disclosure their personal performance rates to patients. The main ethical question addressed is what type of information surgeons owe their patients during informed consent. This question comprises 3 related, ethically relevant considerations that are explored in detail: 1) Does surgeon-specific performance information enhance patient decision-making? 2) Do patients want this type of information? 3) How do the potential benefits of disclosure balance against the risks? RESULTS: Calculating individual performance measures requires tradeoffs and involves inherent uncertainty. There is a lack of evidence regarding whether patients want this information, whether it facilitates their decision-making for surgery, and how it is best communicated to them. Disclosure of personal performance rates during informed consent has the potential benefits of enhancing patient autonomy, improving patient decision-making, and improving quality of care. The major risks of disclosure include inaccurate and misleading performance rates, avoidance of high-risk cases, unjust damage to surgeon's reputations, and jeopardized patient trust. CONCLUSION: At this time, we think that, for most conditions, surgical procedures, and outcomes, the accuracy of surgeon- and patient-specific performance rates is illusory, obviating the ethical obligation to communicate them as part of the informed consent process. Nonetheless, the surgical profession has the duty to develop information systems that allow for performance to be evaluated to a high degree of accuracy. In the meantime, patients should be informed of the quantity of procedures their surgeons have performed, providing an idea of the surgeon's experience and qualitative idea of potential risk. PMID- 17414596 TI - The process of informed consent: neither the time nor the place for disclosure of surgeon-specific outcomes. PMID- 17414597 TI - Long-term outcomes of performing a postdoctoral research fellowship during general surgery residency. AB - OBJECTIVE: To determine whether dedicated research time during surgical residency leads to funding following postgraduate training. SUMMARY BACKGROUND DATA: Unlike other medical specialties, a significant number of general surgery residents spend 1 to 3 years in dedicated laboratory research during their training. The impact this has on obtaining peer reviewed research funding after residency is unknown. METHODS: Survey of all graduates of an academic general surgery resident program from 1990 to 2005 (n = 105). RESULTS: Seventy-five (71%) of survey recipients responded, of which 66 performed protected research during residency. Fifty-one currently perform research (mean effort, 26%; range, 2%-75%). Twenty three respondents who performed research during residency (35%) subsequently received independent faculty funding. Thirteen respondents (20%) obtained NIH grants following residency training. The number of papers authored during resident research was associated with obtaining subsequent faculty grant support (9.3 vs. 5.2, P = 0.02). Faculty funding was associated with obtaining independent research support during residency (42% vs. 17%, P = 0.04). NIH-funded respondents spent more combined years in research before and during residency (3.7 vs. 2.8, P = 0.02). Academic surgeons rated research fellowships more relevant to their current job than private practitioners (4.3 vs. 3.4 by Likert scale, P < 0.05). Both groups considered research a worthwhile use of their time during residency (4.5 vs. 4.1, P = not significant). CONCLUSIONS: A large number of surgical trainees who perform a research fellowship in the middle of residency subsequently become funded investigators in this single-center survey. The likelihood of obtaining funding after residency is related to productivity and obtaining grant support during residency as well as cumulative years of research prior to obtaining a faculty position. PMID- 17414598 TI - Surgical research promotes world peace. PMID- 17414599 TI - Achieving the National Quality Forum's "Never Events": prevention of wrong site, wrong procedure, and wrong patient operations. AB - OBJECTIVE: Review the evidence regarding methods to prevent wrong site operations and present a framework that healthcare organizations can use to evaluate whether they have reduced the probability of wrong site, wrong procedure, and wrong patient operations. SUMMARY BACKGROUND DATA: Operations involving the wrong site, patient, and procedure continue despite national efforts by regulators and professional organizations. Little is known about effective policies to reduce these "never events," and healthcare professional's knowledge or appropriate use of these policies to mitigate events. METHODS: A literature review of the evidence was performed using PubMed and Google; key words used were wrong site surgery, wrong side surgery, wrong patient surgery, and wrong procedure surgery. The framework to evaluate safety includes assessing if a behaviorally specific policy or procedure exists, whether staff knows about the policy, and whether the policy is being used appropriately. RESULTS: Higher-level policies or programs have been implemented by the American Academy of Orthopaedic Surgery, Joint Commission on Accreditation of Healthcare Organizations, Veteran's Health Administration, Canadian Orthopaedic, and the North American Spine Society Associations to reduce wrong site surgery. No scientific evidence is available to guide hospitals in evaluating whether they have an effective policy, and whether staff know of the policy and appropriately use the policy to prevent "never events." CONCLUSIONS: There is limited evidence of behavioral interventions to reduce wrong site, patient, and surgical procedures. We have outlined a framework of measures that healthcare organizations can use to start evaluating whether they have reduced adverse events in operations. PMID- 17414600 TI - A novel approach to high definition, high-contrast video capture in abdominal surgery. AB - OBJECTIVE: The aim of this study was to define the best available option for video capture of surgical procedures for educational and archival purposes, with a view to identifying methods of capturing high-quality footage and identifying common pitfalls. SUMMARY BACKGROUND DATA: Several options exist for those who wish to record operative surgical techniques on video. While high-end equipment is an unnecessary expense for most surgical units, several techniques are readily available that do not require industrial-grade audiovisual recording facilities, but not all are suited to every surgical application. METHODS: We surveyed and evaluated the available technology for video capture in surgery. Our evaluation included analyses of video resolution, depth of field, contrast, exposure, image stability, and frame composition, as well as considerations of cost, accessibility, utility, feasibility, and economies of scale. RESULTS: Several video capture options were identified, and the strengths and shortcomings of each were catalogued. None of the commercially available options was deemed suitable for high-quality video capture of abdominal surgical procedures. A novel application of off-the-shelf technology was devised to address these issues. CONCLUSIONS: Excellent quality video capture of surgical procedures within deep body cavities is feasible using commonly available equipment and technology, with minimal technical difficulty. PMID- 17414601 TI - Application of fibrin glue sealant after hepatectomy does not seem justified: results of a randomized study in 300 patients. AB - OBJECTIVE: To evaluate the efficacy, amount of hemorrhage, biliary leakage, complications, and postoperative evolution after fibrin glue sealant application in patients undergoing liver resection. SUMMARY BACKGROUND DATA: Fibrin sealants have become popular as a means of improving perioperative hemostasis and reducing biliary leakage after liver surgery. However, trials regarding its use in liver surgery remain limited and of poor methodologic quality. PATIENTS AND METHODS: A total of 300 patients undergoing hepatic resection were randomly assigned to fibrin glue application or control groups. Characteristics and debit of drainage and postoperative complications were evaluated. The amount of blood loss, measurements of hematologic parameters liver test, and postoperative evolution (particularly involving biliary fistula and morbidity) was also recorded. RESULTS: Postoperatively, no differences were observed in the amount of transfusion (0.15 +/- 0.66 vs. 0.17 +/- 0.63 PRCU; P = 0.7234) or in the patients that required transfusion (18% vs. 12%; P = 0.2), respectively, for the fibrin glue or control group. There were no differences in overall drainage volumes (1180 +/- 2528 vs. 960 +/- 1253 mL) or in days of postoperative drainage (7.9 +/- 5 vs. 7.1 +/- 4.7). Incidence of biliary fistula was similar in the fibrin glue and control groups, (10% vs. 11%). There were no differences regarding postoperative morbidity between groups (23% vs. 23%; P = 1). CONCLUSIONS: Application of fibrin sealant in the raw surface of the liver does not seem justified. Blood loss, transfusion, incidence of biliary fistula, and outcome are comparable to patients without fibrin glue. Therefore, discontinuation of routine use of fibrin sealant would result in significant cost saving. PMID- 17414602 TI - The ratio between metastatic and examined lymph nodes (N ratio) is an independent prognostic factor in gastric cancer regardless of the type of lymphadenectomy: results from an Italian multicentric study in 1853 patients. AB - PURPOSE: To investigate whether the ratio between metastatic and examined lymph nodes (N ratio) is a better prognostic factor as compared with traditional staging systems in patients with gastric cancer regardless of the extension of lymph node dissection. PATIENTS & METHODS: We retrospectively reviewed the data of 1853 patients who underwent radical resection for gastric carcinoma at 6 Italian centers. Patients with >15 (group 1, n = 1421) and those with 25%) were determined by the best cut-off approach. RESULTS: After a median follow-up of 45.5 months (range, 4-182 months), the 5-year overall survival of N0, N1, and N2 patients of group 1 versus group 2 was 83.4% versus 74.2% (P = 0.0026), 54.3% versus 44.3% (P = 0.018), and 32.7% versus 14.7% (P = 0.004), respectively, suggesting that a low number of excised lymph nodes can lead to the understaging of patients. N ratio identified subsets of patients with significantly different survival rates within N1 and N2 stages in both groups. At multivariate analysis, the N ratio (but not N stage) was retained as an independent prognostic factor both in group 1 and group 2 (HR for N ratio 1, N ratio 2, and N ratio 3 = 1.67, 2.96, and 6.59, and 1.56, 2.68, and 4.28, respectively). In our series, the implementation of N ratio led to the identification of subgroups of patients prognostically more homogeneous than those classified by the TNM system. CONCLUSION: N ratio is a simple and reproducible prognostic tool that can stratify patients with gastric cancer also in case of limited lymph node dissection. These data may represent the rational for improving the prognostic power of current UICC TNM staging system and ultimately the selection of patients who may most benefit from adjuvant treatments. PMID- 17414603 TI - Postoperative irradiation for rectal cancer increases the risk of small bowel obstruction after surgery. AB - OBJECTIVE: To determine the risk of small bowel obstruction (SBO) after irradiation (RT) for rectal cancer BACKGROUND: : SBO is a frequent complication after standard resection of rectal cancer. Although the use of RT is increasing, the effect of RT on risk of SBO is unknown. METHODS: We conducted a retrospective cohort study using Surveillance, Epidemiology, and End Results cancer registry data linked to Medicare claims data to determine the effect of RT on risk of SBO. Patients 65 years of age and older diagnosed with nonmetastatic invasive rectal cancer treated with standard resection from 1986 through 1999 were included. We determined whether patients had undergone RT and evaluated the effect of RT and timing of RT on the incidence of admission to hospital for SBO, adjusting for potential confounders using a proportional hazards model. RESULTS: We identified a total of 5606 patients who met our selection criteria: 1994 (36%) underwent RT, 74% postoperatively. Patients were followed for a mean of 3.8 years. A total of 614 patients were admitted for SBO over the study period; 15% of patients in the RT group and 9% of patients in the nonirradiated group (P < 0.001). After controlling for age, sex, race, diagnosis year, type of surgery, and stage, we found that patients who underwent postoperative RT were at higher risk of SBO, hazard ratio 1.69 (95% CI, 1.3-2.1). However, the long-term risk associated with preoperative irradiation was not statistically significant (hazard ratio, 0.89; 95% CI, 0.55-1.46). CONCLUSIONS: Postoperative but not preoperative RT after standard resection of rectal cancer results in an increased risk of SBO over time. PMID- 17414604 TI - Single-amplicon MSH2 A636P mutation testing in Ashkenazi Jewish patients with colorectal cancer: role in presurgical management. AB - OBJECTIVE: This study summarizes our initial experience with prospective, single amplicon (mutation-specific) A636P testing in Ashkenazi Jewish patients at risk for Hereditary Nonpolyposis Colorectal Cancer (HNPCC). SUMMARY BACKGROUND DATA: We previously described a founder mutation, MSH2*1906G >C (A636P) that causes HNPCC in 8/1345 (0.59%) of Ashkenazim with colorectal cancer. The mutation was more common in Ashkenazim diagnosed at 9 months) from resection to recurrence were more likely to benefit from reresection compared with patients with recurrence within 9 months (median survival 17.0 vs. 7.4 months; P = 0.004). The in-hospital morbidity and mortality rate of resected patients was 20% and 6.7% compared with 13.3% and 0% of patients who underwent only exploration/palliative bypass. CONCLUSION: Resection for recurrent pancreatic cancer can be carried out safely. Further studies are required to address the question whether a subgroup of patients might actually benefit from this procedure. PMID- 17414606 TI - Distal pancreatectomy: risk factors for surgical failure in 302 consecutive cases. AB - OBJECTIVE: The objective of this study was to identify potential risk factors for mortality and morbidity after distal pancreatectomy, with special focus on the formation of pancreatic fistula. SUMMARY BACKGROUND DATA: Distal pancreatectomy can be performed with low mortality and acceptable morbidity rates. Pancreatic fistulas, occurring in 10% to 20% of cases, remain a problem that contributes significantly to morbidity, length of stay, and overall costs. METHODS: From November 1993 to February 2006, perioperative and postoperative data of 302 consecutive patients were recorded. Univariate and multivariate analyses of potential risk factors for morbidity and for the formation of pancreatic fistula were performed. The surgical techniques used for closure were categorized into 4 groups: 1) anastomosis, 2) seromuscular patch, 3) closure by suture, and 4) closure using a stapling device. RESULTS: Indications for resection were pancreatic tumors in 62% of patients, nonpancreatic tumors in 23%, chronic pancreatitis in 12%, and others in 3%. The spleen was preserved in 24% of patients. The morbidity and mortality rates for distal pancreatectomy in this series were 35% and 2%, respectively. The prevalence of pancreatic fistula was 12%. Univariate and multivariate analyses indicated that closure using a stapling device and an operating time >or=480 minutes were associated with a higher incidence of pancreatic fistula (odds ratio = 2.6 and 4.2, respectively). Overall morbidity was mainly influenced by the extent of resection (multivisceral vs. conventional; odds ratio = 1.7). CONCLUSION: Pancreatic leak remains a common complication after distal pancreatectomy. Our series suggests that stapler closure of the pancreatic remnant is associated with a significantly higher fistula rate. PMID- 17414607 TI - Preoperative hepatic and regional arterial chemotherapy in the prevention of liver metastasis after colorectal cancer surgery. AB - OBJECTIVE: To investigate whether preoperative hepatic and regional arterial chemotherapy is able to prevent liver metastasis and improve overall survival in patients receiving curative colorectal cancer resection. METHODS: Patients with stage II or stage III colorectal cancer (CRC) were randomly assigned to receive preoperative hepatic and regional arterial chemotherapy (PHRAC group, n = 110) or surgery alone (control group, n = 112). The primary endpoint was disease-free survival, whereas the secondary endpoints included liver metastasis-free survival and overall survival. RESULTS: There were no significant differences in overall morbidity between PHRAC and Control groups. During the follow-up period (median, 36 months), the median liver metastasis time for patients with stage III CRC was significantly longer in the PHRAC group (16 +/- 3 months vs. 8 +/- 1 months, P = 0.01). In stage III patients, there was also significant difference between the 2 groups with regard to the incidence of liver metastasis (20.6% vs. 28.3%, P = 0.03), 3-year disease-free survival (74.6% vs. 58.1%, P = 0.0096), 3-year overall survival (87.7% vs. 75.7%, P = 0.020), and the median survival time (40.1 +/- 4.6 months vs. 36.3 +/- 3.2 months, P = 0.03). In the PHRAC arm, the risk ratio of recurrence was 0.61 (95% CI, 0.51-0.79, P = 0.0002), of death was 0.51 (95% CI, 0.32-0.67; P = 0.009), and of liver metastasis was 0.73 (95% CI, 0.52-0.86; P = 0.02). In contrast, PHRAC seemed to be no benefit for stage II patients. Toxicities, such as hepatic toxicity and leukocyte decreasing, were mild and could be cured with medicine. CONCLUSIONS: Preoperative hepatic and regional arterial chemotherapy, in combination with surgical resection, could be able to reduce and delay the occurrence of liver metastasis and therefore improve survival rate in patients with stage III colorectal cancer. PMID- 17414608 TI - Long-term results of hyperthermic, isolated limb perfusion for melanoma: a reflection of tumor biology. AB - PURPOSE: To review the long-term duration of limb tumor complete remission (CR) and patient survival following therapeutic hyperthermic isolated limb perfusion (ILP) with cytotoxic drugs for melanoma. METHODS: A retrospective case series of 124 ILPs performed in 111 patients. RESULTS: There were 120 assessable ILPs. Patient staging (M.D. Anderson system) was stage II 11.7%, stage IIIA 44.2%, stage IIIAB 33.3%, and stage IV 10.8%. CR was initially attained after 83 ILPs (69.2%) and partial remission (PR) after 19 ILPs (15.8%). Limb CR was maintained in 28 (33.7%) of the 83 cases. Disease recurred in the perfused limb after an initial CR in the remaining 55 cases (median time to recurrence, 11 months); in 19 of these cases, the limb was disease-free at last follow-up after further locoregional treatment. A long-term CR was achieved, with or without further treatment, in 47 (56.6%) of the 83 cases in which an initial CR had occurred (mean follow-up, 97 months; median, 65 months). There was no significant difference in long-term local remission for stage IIIA and IIIAB patients. Five year survival for those who had a partial or no response to ILP was 7%. Ten-year survival for those who had a long-term CR was 49%. CONCLUSIONS: ILP, with or without further locoregional treatment, achieved long-term control of recurrent and metastatic limb disease in 56.6% of cases in which an initial CR was achieved. A complete response to ILP was a positive prognostic indicator for survival, probably reflecting more favorable tumor biology in this subset of patients. PMID- 17414609 TI - Iterative cytoreductive surgery associated with hyperthermic intraperitoneal chemotherapy for treatment of peritoneal carcinomatosis of colorectal origin with or without liver metastases. AB - INTRODUCTION: The aim of this study was to evaluate the results of an aggressive strategy in patients presenting peritoneal carcinomatosis (PC) from colorectal cancer with or without liver metastases (LMs) treated with cytoreductive surgery (CS) and hyperthermic intraperitoneal chemotherapy (HIPEC). PATIENTS AND METHODS: The population included 43 patients who had 54 CS+HIPEC for colorectal PC from 1996 to 2006. Sixteen patients (37%) presented LMs. Eleven patients (25%) presented occlusion at the time of PC diagnosis. Ascites was present in 12 patients (28%). Seventy-seven percent of the patients were Gilly 3 (diffuse nodules, 5-20 mm) and Gilly 4 (diffuse nodules>20 mm). The main endpoints were morbidity, mortality, completeness of cancer resection (CCR), and actuarial survival rates. RESULTS: The CS was considered as CCR-0 (no residual nodules) or CCR-1 (residual nodules <5 mm) in 30 patients (70%). Iterative procedures were performed in 26% of patients. Three patients had prior to CS + HIPEC, 10 had concomitant minor liver resection, and 3 had differed liver resections (2 right hepatectomies) 2 months after CS + HIPEC. The mortality rate was 2.3% (1 patient). Seventeen patients (39%) presented one or multiple complications (per procedure morbidity = 31%). Complications included deep abscess (n = 6), wound infection (n = 5), pleural effusion (n = 5), digestive fistula (n = 4), delayed gastric emptying syndrome (n = 4), and renal failure (n = 3). Two patients (3.6%) were reoperated. The median survival was 38.4 months (CI, 32.8-43.9). Actuarial 2 and 4-year survival rates were 72% and 44%, respectively. The survival rates were not significantly different between patients who had CS + HIPEC for PC alone (including the primary resection) versus those who had associated LMs resection (median survival, 35.3 versus 36.0 months, P = 0.73). CONCLUSION: Iterative CS + HIPEC is an effective treatment in PC from colorectal cancer. The presence of resectable LMs associated with PC does not contraindicate the prospect of an oncologic treatment in these patients. PMID- 17414610 TI - Total thyroidectomy plus neck dissection in differentiated papillary thyroid carcinoma patients: pattern of nodal metastasis, morbidity, recurrence, and postoperative levels of serum parathyroid hormone. AB - OBJECTIVE: To investigate the pattern of nodal metastasis, morbidity, recurrence rates of papillary thyroid carcinoma (PTC), and parathyroid hormone (PTH) responses following neck dissection (ND) plus total thyroidectomy (TT). SUMMARY BACKGROUND DATA: While hypoparathyroidism develops after TT plus ND, little is known of postoperative PTH response. METHODS: Of 155 PTC patients, 82 underwent TT plus bilateral central ND with/without lateral ND, while 73 underwent TT alone. The nodal metastasis pattern was determined and the recurrence, morbidity, and postoperative levels of serum calcium and PTH were compared between 2 groups. RESULTS: Of the 82 node dissection patients, metastatic nodes were present in the central neck of 51 (62.2%) and the lateral neck of 21 (25.6%) patients, most frequently in the ipsilateral and pretracheal central nodes and lateral jugular nodes. Four regional recurrences (2.6%) were found in 3 patients of the no node dissection group and one of the node dissection group (P = 0.37) during the follow-up lasting a mean 52 months. Overall morbidity and hypocalcemia was higher in the node dissection group than the no node dissection group (41 of 82, 50%; vs. 9 of 73, 12.3%; P < 0.001; 25 of 82, 30.5%; vs. 7 of 73, 9.6%; P = 0.001). Serum PTH levels significantly decreased immediately postoperatively in the node dissection group and remained low for several weeks thereafter. CONCLUSIONS: Serum PTH levels were significantly reduced following ND in PTC patients. Our data suggest that, when performing therapeutic ND plus TT, particular effort should be made to preserve the parathyroid glands and to monitor their function. PMID- 17414611 TI - Gene expression profiles differentiate between sterile SIRS and early sepsis. AB - INTRODUCTION: The systemic inflammatory response syndrome (SIRS) occurs frequently in critically ill patients and presents similar clinical appearances despite diverse infectious and noninfectious etiologies. Despite similar phenotypic expression, these diverse SIRS etiologies may induce divergent genotypic expressions. We hypothesized that gene expression differences are present between sepsis and uninfected SIRS prior to the clinical appearance of sepsis. METHODS: Critically ill uninfected SIRS patients were followed longitudinally for the development of sepsis. All patients had whole blood collected daily for gene expression analysis by Affymetrix Hg_U133 2.0 Plus microarrays. SIRS patients developing sepsis were compared with those remaining uninfected for differences in gene expression at study entry and daily for 3 days prior to conversion to sepsis. Acceptance criteria for differentially expressed genes required: >1.2 median fold change between groups and significance on univariate and multivariate analysis. Differentially expressed genes were annotated to pathways using DAVID 2.0/EASE analysis. RESULTS: A total of 12,782 (23.4%) gene probes were differentially expressed on univariate analysis 0 to 48 hours before clinical sepsis. 626 (1.1%) probes met acceptance criteria, corresponding to 459 unique genes, 65 (14.2%) down and 395 (85.8%) up expressed. These genes annotated to 10 pathways that functionally categorized to 4 themes involving innate immunity, cytokine receptors, T cell differentiation, and protein synthesis regulation. CONCLUSIONS: Sepsis has a unique gene expression profile that is different from uninfected inflammation and becomes apparent prior to expression of the clinical sepsis phenotype. PMID- 17414612 TI - The association between fluid administration and outcome following major burn: a multicenter study. AB - OBJECTIVE: To determine patient and injury variables that influence fluid requirements following burn injury and examine the association between fluid volume received and outcome. BACKGROUND: Fluid resuscitation remains the cornerstone of acute burn management. Recent studies suggest that patients today are receiving more fluid per percent total body surface area (TBSA) than in the past. Therefore, there is a need to better define the factors that impact fluid requirements and to determine the effects of fluid volumes on outcome. METHODS: This study was part of a federally funded multicenter study. Multilinear regression analyses were performed to determine the patient and injury characteristics that most influenced fluid resuscitation volumes received. To assess the association of fluid volumes on outcome, propensity scores were developed to provide a predicted volume of fluid for each patient. Logistic models were then used to assess the impact of excess fluid beyond predicted volumes on outcome. RESULTS: Seventy-two patients were included in this analysis. Average patient age was 40.6 years and average TBSA was 44.5%. Average fluid volume received during the first 24 hours after injury was 5.2/mL/kg/TBSA. Significant predictors of fluid received included % TBSA, age, intubation status, and weight. Increased fluid volume received increased risk of development of pneumonia (odds ratio [OR] = 1.92), bloodstream infections (OR =2.33), adult respiratory distress syndrome (OR = 1.55), multiorgan failure (OR= 1.49), and death (OR = 1.74). CONCLUSION: TBSA, age, weight, and intubation status on admission were significant predictors of fluid received. Patients who received larger volumes of resuscitation fluid were at higher risk for injury complications and death. PMID- 17414613 TI - The effects of preexisting medical comorbidities on mortality and length of hospital stay in acute burn injury: evidence from a national sample of 31,338 adult patients. AB - OBJECTIVE: To determine whether and to what extent preexisting medical comorbidities influence mortality risk and length of hospitalization in patients with acute burn injury. SUMMARY BACKGROUND DATA: The effects on mortality and length of stay of a number of important medical comorbidities have not been examined in acute burn injury. Existing studies that have investigated the effects of medical comorbidities on outcomes in acute burn injury have produced inconsistent results, chiefly due to the use of relatively small samples from single burn centers. METHODS: Records of 31,338 adults who were admitted with acute burn injury to 70 burn centers from the American Burn Association National Burn Repository, were reviewed. A burn-specific list of medical comorbidities was derived from diagnoses included in the Charlson Index of Comorbidities and the Elixhauser method of comorbidity measurement. Logistic regression was used to assess the effects of preexisting medical conditions on mortality, controlling for demographic and burn injury characteristics. Ordinal least squares regression with a logarithmic transformation of the dependent variable was used to assess the relationship of comorbidities with length of stay. RESULTS: In-hospital mortality was significantly predicted by HIV/AIDS (odds ratio [OR] = 10.2), renal disease (OR = 5.1), liver disease (OR = 4.8), metastatic cancer (OR = 4.6), pulmonary circulation disorders (OR = 2.9), congestive heart failure (OR = 2.4), obesity (OR = 2.1), non-metastatic malignancies (OR = 2.1), peripheral vascular disorders (OR = 1.8), alcohol abuse (OR = 1.8), neurological disorders (OR = 1.6), and cardiac arrhythmias (OR = 1.5). Increased length of hospital stay among survivors was significantly predicted by paralysis (90% increase), dementia (60%), peptic ulcer disease (53%), other neurological disorders (52%), HIV/AIDS (49%), renal disease (44%), a psychiatric diagnosis (42%), cerebrovascular disease (41%), cardiac arrhythmias (40%), peripheral vascular disorders (39%), alcohol abuse (36%), valvular disease (32%), liver disease (30%), diabetes (26%), congestive heart failure (23%), drug abuse (20%), and hypertension (17%). CONCLUSIONS: A number of preexisting medical conditions influence outcomes in acute burn injury. Patients with preburn HIV/AIDS, metastatic cancer, liver disease, and renal disease have particularly poor prognoses. PMID- 17414614 TI - Hypertonic resuscitation modulates the inflammatory response in patients with traumatic hemorrhagic shock. AB - OBJECTIVE: To determine the effect of resuscitation with hypertonic saline/dextran (HSD) on the innate immune response after injury. SUMMARY OF BACKGROUND DATA: Hypovolemic shock causes a whole body ischemia/reperfusion injury, leading to dysregulation of the inflammatory response and multiple organ dysfunction syndrome. Hypertonicity has been shown to modulate the innate immune response in vitro and in animal models of hemorrhagic shock, but the effect on the inflammatory response in humans is largely unknown. METHODS: Serial blood samples were drawn (12, 24, 72 hours and 7 days after injury) from patients enrolled in a prospective, randomized, double-blind trial of traumatic hypovolemic shock, HSD (250 mL) versus lactated Ringer's solution (LR) as the initial resuscitation fluid. Neutrophil (PMN) CD11b/CD18 expression was assessed via whole blood FACS analysis with and without stimulation (fMLP 5 micromol/L or PMA 5 micromol/L). PMN respiratory burst was assessed using the nitro-blue tetrazolium assay. Monocytes stimulated with 100 ng LPS for 18 hours were assessed for cytokine production (TNF-alpha, IL-1Beta, IL-6, IL-10, IL-12). RESULTS: Sixty-two patients (36 HSD, 26 LR) and 20 healthy volunteers were enrolled. CD11b expression, 12 hours after injury, was increased 1.5-fold in patients resuscitated with LR compared with controls. Those resuscitated with HSD had a significant reduction in CD11b expression 12 hours after injury, compared with LR. There was no difference in respiratory burst early after injury. Monocytes from injured patients expressed lower levels of all cytokines in comparison to normal controls. Patients give HSD showed a trend toward higher levels of IL-1beta and IL10 production in response to LPS, 12 hours after injury. CONCLUSION: HSD resuscitation results in transient inhibition of PMN CD11b expression and partial restoration of the normal monocyte phenotype early after injury. PMID- 17414615 TI - Nutritional route affects ERK phosphorylation and cytokine production in hepatic mononuclear cells. AB - OBJECTIVE: To clarify the influence of nutritional route on hepatic immunity in a murine model. SUMMARY BACKGROUND DATA: Parenteral nutrition is disadvantageous for preventing infectious complications in critically ill and/or severely injured patients as compared with enteral nutrition. To date, lack of enteral nutrition has been demonstrated to impair mucosal immunity, gut barrier function, and the peritoneal defense system. However, influences of nutritional route on hepatic immunity, another important defense system against infection, have not been well studied. METHODS: Male ICR mice were randomized to 3 groups: ad libitum chow (chow), intravenous (IV)-TPN and intragastric (IG)-TPN groups. The TPN groups were given isocaloric and isonitrogenous TPN solutions. After the mice had been fed for 5 days, hepatic mononuclear cells (MNCs) were isolated. Hepatic MNC numbers and functions (cytokine production, intracellular signaling, and LPS receptor expression) were determined. Moreover, 1.0 x 10 Pseudomonas aeruginosa were delivered by intraportal injection. Survival and histology were examined. RESULTS: Hepatic MNC numbers were significantly lower in the IV-TPN group than in the chow and IG-TPN groups, without subpopulation changes. As compared with enterally fed mice, cytokine production (TNF-alpha, IFN-gamma, and IL-10) by hepatic MNCs in response to LPS was impaired in parenterally fed mice in association with blunted phosphorylation of ERK1/2, a MAPK. Hepatic MNCs from IV TPN mice showed decreased expressions of CD14 and TLR4/MD2, as compared with enterally fed mice. Survival times were reduced in the IV-TPN group as compared with the chow and IG-TPN groups. CONCLUSION: Preservation of hepatic immunity with enteral feeding is important for prevention of infectious complications in severely injured and/or critically ill patients. PMID- 17414616 TI - Effect of alcohol on Glasgow Coma Scale in head-injured patients. AB - OBJECTIVE: Almost 50% of traumatic brain-injured (TBI) patients are alcohol intoxicated. The Glasgow Coma Scale (GCS) is frequently used to direct diagnostic and therapeutic decisions in these patients. It is commonly assumed that alcohol intoxication reduces GCS, thus limiting its utility in intoxicated patients. The purpose of this study was to test the hypothesis that the presence of blood alcohol has a clinically significant impact on GCS in TBI patients. METHODS: The National Trauma Data Bank of the American College of Surgeons was queried (1994 2003). Patients 18 to 45 years of age with blunt injury mechanism, whose GCS in the emergency department, survival status, anatomic severity of TBI (Head Abbreviated Injury Score [AIS]), and blood alcohol testing status were known, were included. GCS of patients who tested positive for alcohol (n = 55,732) was compared with GCS of patients who tested negative (n = 53,197), stratified by head AIS. RESULTS: Groups were similar in age (31 +/- 8 vs. 30 +/- 8 years), Injury Severity Score (ISS; 12 +/- 11 vs. 12 +/- 11), systolic blood pressure in the ED (131 +/- 25 vs. 134 +/- 25 mm Hg), TRISS (Trauma Injury Severity Score; probability of survival (94% +/- 16% vs. 95% +/- 15%), and actual survival (96% vs. 96%). When stratified by anatomic severity of TBI, the presence of alcohol did not lower GCS by more than 1 point in any head AIS group (GCS in alcohol positive vs. alcohol-negative patients; AIS 1 = 13.9 +/- 2.8 vs. 14.3 +/- 2.3; AIS 2 = 13.4 +/- 3.2 vs. 14.1 +/- 2.4; AIS 3 = 11.1 +/- 4.7 vs. 11.6 +/- 4.6; AIS 4 = 9.8 +/- 4.9 vs. 10.4 +/- 4.9; AIS 5 = 5.5 +/- 3.8 vs. 5.9 +/- 4.1, AIS 6: 3.4 +/- 1.1 vs. 3.8 +/- 2.8). CONCLUSION: Alcohol use does not result in a clinically significant reduction in GCS in trauma patients. Attributing low GCS to alcohol intoxication in TBI patients may delay necessary diagnostic and therapeutic interventions. PMID- 17414617 TI - Mortality after groin hernia surgery. AB - OBJECTIVE: To analyze mortality following groin hernia operations. SUMMARY BACKGROUND DATA: It is well known that the incidence of groin hernia in men exceeds the incidence in women by a factor of 10. However, gender differences in mortality following groin hernia surgery have not been explored in detail. METHODS: The study comprises all patients 15 years or older who underwent groin hernia repair between January 1, 1992 and December 31, 2005 at units participating in the Swedish Hernia Register (SHR). Postoperative mortality was defined as standardized mortality ratio (SMR) within 30 days, ie, observed deaths of operated patients over expected deaths considering age and gender of the population in Sweden. RESULTS: A total of 107,838 groin hernia repairs (103,710 operations), were recorded prospectively. Of 104,911 inguinal hernias, 5280 (5.1%) were treated emergently, as compared with 1068 (36.5%) of 2927 femoral hernias. Femoral hernia operations comprised 1.1% of groin hernia operations on men and 22.4% of operations on women. After femoral hernia operation, the mortality risk was increased 7-fold for both men and women. Mortality risk was not raised above that of the background population for elective groin hernia repair, but it was increased 7-fold after emergency operations and 20-fold if bowel resection was undertaken. Overall SMR was 1.4 (95% confidence interval, 1.2 1.6) for men and 4.2 (95% confidence interval, 3.2-5.4) for women, in accordance with a greater proportion of emergency operations among women compared with men, 17.0%, versus 5.1%. CONCLUSIONS: Mortality risk following elective hernia repair is low, even at high age. An emergency operation for groin hernia carries a substantial mortality risk. After groin hernia repair, women have a higher mortality risk than men due to a greater risk for emergency procedure irrespective of hernia anatomy and a greater proportion of femoral hernia. PMID- 17414618 TI - Should preoperative mechanical bowel preparation be abandoned? PMID- 17414619 TI - Prophylactic ilioinguinal neurectomy in open inguinal hernia repair. PMID- 17414620 TI - Gene expression profiles as biomarkers for the prediction of chemotherapy drug response in human tumour cells. AB - Genome profiling approaches such as cDNA microarray analysis and quantitative reverse transcription polymerase chain reaction are playing ever-increasing roles in the classification of human cancers and in the discovery of biomarkers for the prediction of prognosis in cancer patients. Increasing research efforts are also being directed at identifying set of genes whose expression can be correlated with response to specific drugs or drug combinations. Such genes hold the prospect of tailoring chemotherapy regimens to the individual patient, based on tumour or host gene expression profiles. This review outlines recent advances and challenges in using genome profiling for the identification of tumour or host genes whose expression correlates with response to chemotherapy drugs both in vitro and in clinical studies. Genetic predictors of response to a variety of anticancer agents are discussed, including the anthracyclines, taxanes, topoisomerase I and II inhibitors, nucleoside analogs, alkylating agents, and vinca alkaloids. PMID- 17414621 TI - Peroxisome proliferator-activated receptor-gamma antagonists exhibit potent antiproliferative effects versus many hematopoietic and epithelial cancer cell lines. AB - Peroxisome proliferator-activated receptor-gamma ligands have preclinical and clinical anticancer activity. Most studies in this area address agonists, with relatively few reports on anticancer effects of peroxisome proliferator-activated receptor-gamma antagonists. Thus, we evaluated the two pure peroxisome proliferator-activated receptor-gamma antagonists, T0070907 and GW9662, on a panel of hematopoietic and epithelial cell lines. The peroxisome proliferator activated receptor-gamma antagonists and a reference agonist (pioglitazone) were tested in an in-vitro proliferation assay on a panel of seven hematopoietic and nine epithelial cancer cell lines, some of which are chemoresistant. Peroxisome proliferator-activated receptor-gamma expression was measured by immunoblotting, as was the effect of treatment with these agents on peroxisome proliferator activated receptor-gamma levels. The effect of exogenous interleukin-6, an antiapoptotic cytokine, on growth inhibition was evaluated as well as the apoptotic effects of these drugs. The peroxisome proliferator-activated receptor gamma antagonists showed significantly greater potency on all cell lines (IC50s of 3.2-29.7 versus 26.5-78.7 micromol/l for pioglitazone) and greater maximum growth inhibition. Peroxisome proliferator-activated receptor-gamma levels did not correlate with growth inhibition in this panel of cell lines. Combinations of peroxisome proliferator-activated receptor-gamma antagonists and the agonist actually showed schedule-dependent increases in growth inhibition. Exogenous interleukin-6 did not induce resistance to these agents. Both the antagonists and the agonist induced apoptosis, but only the former drugs showed caspase dependence. These two peroxisome proliferator-activated receptor-gamma antagonists have significantly more potent in-vitro antiproliferative effects versus hematopoietic and epithelial cancer cell lines. This effect does not correlate with peroxisome proliferator-activated receptor-gamma levels, suggesting alternative mechanisms or other targets of action. These findings support further translational studies to explore the mechanism of action and therapeutic potential of this class of agents. PMID- 17414622 TI - Leukotriene B4 receptor antagonist LY293111 induces S-phase cell cycle arrest and apoptosis in human pancreatic cancer cells. AB - We have previously shown that the leukotriene B4 receptor antagonist, LY293111 inhibits proliferation and induces apoptosis in human pancreatic cancer cells both in vitro and in vivo. In the current study, we investigated the molecular mechanisms of LY293111-induced apoptosis and cell cycle arrest. Two human pancreatic cancer cell lines were used in this study, MiaPaCa-2 and AsPC-1. Cell cycle analysis by flow cytometry showed a dramatic increase in the percentage of apoptotic cells as well as S-phase arrest after treatment with 250 nmol/l LY293111 for up to 48 h. Western blotting indicated that LY293111 treatment induced cytochrome c release from the mitochondria into the cytosol, accompanied by caspase-9, caspase-7 and caspase-3 activation, and cleavage of poly ADP-ribose polymerase. Caspase-8 was not activated by LY293111. A decrease was found in the expression of the antiapoptotic proteins, Bcl-2 and Mcl-1, and an increase in the proapoptotic protein, Bax. LY293111 reduced the expression of CDK2, cyclin A and cyclin E, consistent with the S-phase arrest observed in these cells. The expression of cyclin-dependent kinase inhibitors, p21 and p27 was not affected by LY293111 treatment. In conclusion, LY293111 induces apoptosis in human pancreatic cancer cells through the mitochondria-mediated pathway. LY293111 also induces S phase arrest with downregulation of CDK2, cyclin A and cyclin E. Blockade of leukotriene B4 metabolic pathway may provide a novel treatment for human pancreatic cancer. PMID- 17414623 TI - Necrotic pathway in human osteosarcoma Saos-2 cell death induced by chloroacetaldehyde. AB - Chloroacetaldehyde, a metabolite of the anticancer drug ifosfamide, may be responsible for serious adverse effects like encephalopathy in ifosfamide chemotherapy. In this study, we demonstrate that chloroacetaldehyde, but not ifosfamide, induces cell death in human osteosarcoma Saos-2 cells and we investigated the mechanism by which this occurs. Chloroacetaldehyde above 30 micromol/l induced significant cell death in a time-dependent manner. Thiol compounds such as N-acetyl cysteine, glutathione and dithiothreitol protected the cells against chloroacetaldehyde-induced cell death, although other nonthiol compounds and the antioxidative enzymes superoxide dismutase and catalase did not, suggesting that reactive oxygen species might not mediate cell death. In cells exposed to chloroacetaldehyde, levels of both total thiols and glutathione were significantly reduced. Chloroacetaldehyde also collapsed the mitochondrial membrane potential of these cells, induced the release of cytochrome c from mitochondria to the cytosol and significantly reduced cellular ATP levels during the course of death. The mitochondrial potential collapse was also prevented by thiol compounds. Flow cytometric analyses by means of annexin-V and propidium iodide double staining and immunofluorescence staining of active caspase-3 revealed that cells subjected to a lethal dose of chloroacetaldehyde displayed features characteristic of necrosis and that caspase-3 was not activated in response to chloroacetaldehyde. Taken together, these findings suggest that Saos 2 cells exposed to chloroacetaldehyde die by necrosis resulting from a decrease in intracellular thiols, disruption of the mitochondrial membrane potential and the depletion of cellular ATP. PMID- 17414624 TI - Induction of apoptosis in human breast adenocarcinoma MCF-7 cells by pterocarnin A from the bark of Pterocarya stenoptera via the Fas-mediated pathway. AB - Pterocarnin A, isolated from the bark of Pterocarya stenoptera (Juylandaceae), was investigated for its antiproliferative activity in human breast adenocarcinoma MCF-7 cells. To identify the anticancer mechanism of pterocarnin A, we assayed its effects on apoptosis, cell cycle distribution, and levels of p53, p21/WAF1, Fas/APO-1 receptor and Fas ligand. The results showed that pterocarnin A induced apoptosis of MCF-7 cells without mediation of p53 and p21/WAF1. We suggest that the Fas/Fas ligand apoptotic system is the main pathway of pterocarnin A-mediated apoptosis of MCF-7 cells. Our study reports here for the first time that the activity of the Fas/Fas ligand apoptotic system may participate in the antiproliferative activity of pterocarnin A in MCF-7 cells. PMID- 17414625 TI - ADH1, an N-cadherin inhibitor, evaluated in preclinical models of angiogenesis and androgen-independent prostate cancer. AB - The conversion from E-cadherin to N-cadherin has been observed in several human cancer types, including prostate cancer, with more homogenous expression of N cadherin detected in high-grade prostate tumors. N-cadherin, in vitro, has been shown to promote cell mobility, migration and invasion of several cancer cell lines, indicating the possibility of N-cadherin as a molecular target of cancer therapy. Herein, we examined the potential of an N-cadherin inhibitor, ADH1, in reducing tumor angiogenesis ex vivo and delaying tumor progression in vivo. Our data demonstrate that ADH1, at the dosages evaluated, does not display either antiangiogenic activity in a rat aortic ring assay or antitumor potential in a PC3 subcutaneous xenograft tumor model. We detected cytotoxic activity in human umbilical vein endothelial cells, PC3, and Tsu-Pr1 cells, when ADH1 exposure was evaluated at 500 micromol/l or above. PMID- 17414626 TI - Antiangiogenic and antitumor activity of a novel vascular endothelial growth factor receptor-2 tyrosine kinase inhibitor ZD6474 in a metastatic human pancreatic tumor model. AB - ZD6474 is a novel, orally available inhibitor of vascular endothelial growth factor receptor kinase insert domain receptor/flk-1 tyrosine kinase activity with additional activity against the epidermal growth factor receptor-1 tyrosine kinase. The aim of this study was to evaluate ZD6474, alone and in combination with gemcitabine, in an orthotopic model of metastatic pancreatic cancer. Nude mice (nine to 10/group) were injected orthotopically with 1x10(6) L3.6pl human pancreatic cancer cells. Eight days later, treatment was initiated with vehicle only, gemcitabine (100 mg/kg intraperitoneal twice weekly), ZD6474 (50 mg/kg oral once daily) or a combination of the two treatments. Animals were killed on day 24 posttreatment initiation. The phosphorylation status level of vascular endothelial growth factor receptor-2 and epidermal growth factor receptor as well as the phosphorylation level of AKT and extracellular signal-regulated kinase-1/2 in different human pancreatic carcinoma cells and in human umbilical vein endothelial cells was analyzed by Western blotting. Compared with controls (1231 mg), the mean weight of treated tumors was reduced to 836, 541 and 308 mg in the gemcitabine, ZD6474 and combination groups, respectively. Lymph node metastasis was significantly reduced in both the ZD6474 alone and combined treatment groups, with 3/10 and 1/5 animals developing metastases, compared with 10/10 and 9/9 in the control and gemcitabine groups (P<0.003 and <0.0003, respectively). Microvessel density and cell proliferation were significantly reduced in the ZD6474 and combined treatment groups (P<0.02). Immunohistochemistry of tumor samples following treatment with ZD6474 resulted in a reduction of the activated and phosphorylated epidermal growth factor receptor, whereas total epidermal growth factor receptor levels were comparable with control tumors. On the basis of Western blot analysis, ZD6474 provides inhibition of tumor angiogenesis through an anti-vascular endothelial growth factor receptor-2 mechanism and inhibition of cancer cell growth through an anti-epidermal growth factor receptor mechanism. ZD6474 decreased primary pancreatic tumor growth and reduced lymph node and liver metastases compared with controls or gemcitabine alone. Tumor growth was inhibited further in animals receiving ZD6474 and gemcitabine in combination. PMID- 17414627 TI - Phase II study of epirubicin plus oxaliplatin and infusional 5-fluorouracil as first-line combination therapy in patients with metastatic or advanced gastric cancer. AB - The purpose of this study was to evaluate the efficacy and safety of an epirubicin, oxaliplatin and infusional 5-fluorouracil combination in patients with advanced gastric cancer. Patients with previously untreated advanced measurable gastric cancer received epirubicin (50 mg/m2, day 1), oxaliplatin (130 mg/m2 2-h infusion, day 1) and 5-fluorouracil (750 mg/m2, 24-h infusion, day 1-3) every 3 weeks. The primary endpoint of this phase II study was the response rate according to Response Evaluation Criteria in Solid Tumors. Out of 48 patients, 46 were evaluable for efficacy and 48 for toxicity. A median of five cycles (range 1 6) was administered. The overall best response rate was 47.8% (95% confidence interval 33-63%) including 2.2% complete responses and 45.6% partial responses. The median time for progression and median overall survival was 5 (95% confidence interval 4.1-5.9) and 11 months (95% confidence interval 8.1-13.9), respectively. Grade 3/4 neutropenia and leukocytopenia were observed in 25 and 12.5% of patients, respectively. Grade 3/4 nonhematological toxicities included nausea (6.3%), vomiting (14.6%), neurological toxicity (10.4%) and mucositis (2.1%). The epirubicin, oxaliplatin and infusional 5-fluorouracil regimen was effective and well tolerated as a front-line chemotherapy for patients with metastatic or advanced gastric cancer, and should be evaluated further. PMID- 17414628 TI - A phase I study of bendamustine hydrochloride administered once every 3 weeks in patients with solid tumors. AB - The present phase I trial was planned to assess the maximum tolerated dose, the dose-limiting toxicity and the pharmacokinetics of bendamustine hydrochloride in a once every 3 weeks schedule, and to recommend a safe dose for future phase II studies. Included were patients with refractory solid tumors. Bendamustine hydrochloride was administered as a short intravenous infusion over 30 min. The starting dose was defined at 160 mg/m2 and dose escalation used increments of 20 mg/m2. Plasma and urine samples were analyzed using validated high-pressure liquid chromatography/fluorescence assays. Twenty-six patients (14 men, 12 women) were enrolled for the study. At 280 mg/m2, one out of four patients developed a thrombocytopenia grade 4, two experienced grade 3 fatigue and three experienced cardiac toxicity (grade 2). The latter toxicity was considered dose limiting also and further dose escalation was stopped. Plasma pharmacokinetics parameters of bendamustine hydrochloride and its metabolites were assessed in 15 patients. Mean pharmacokinetic parameters of bendamustine hydrochloride were a tmax of 32.3 min, a t1/2 of 37.8 min, a volume of distribution of 14.2 l/m and a clearance of 287.8 ml/min/m2. No dose dependency of bendamustine hydrochloride was observed within the used dose range. The metabolites comprised only 23% of the overall area under the concentration-time curve. The maximum tolerated dose of bendamustine hydrochloride on day 1 q 3 weeks is 280 mg/m2. Fatigue and cardiac toxicity were dose limiting. The plasma pharmacokinetics data of bendamustine and its metabolites were in accordance with previous reports. The recommended dose for future trials is 260 mg/m2 every 3 weeks. PMID- 17414629 TI - Chemoimmunotherapy in the treatment of metastatic gastric cancer. AB - Docetaxel, capecitabine and 5-fluorouracil have been shown to be active in the treatment of metastatic gastric adenocarcinoma. Consistent with this finding, the aim of this study was to test this combination in a clinical trial. Forty-one patients with metastatic gastric adenocarcinoma and a median age of 64 years were recruited for the study. The treatment was based on the administration of docetaxel 60 mg/m2 every 4 weeks, leucovorin 200 mg/m2, 5-fluorouracil 400 mg/m2 bolus, and capecitabine 1000 mg/m2 twice daily on days 1 and 2 every 2 weeks. Patients achieving a clinical benefit were treated, as maintenance immunotherapy, with low-dose interleukin-2 and 13-cis-retinoic acid. The primary end point of this phase II study was the response rate. The secondary end points relied on the evaluation of the immunological parameters, toxicity, and progression-free survival and overall survival. The overall response rate in the 41 evaluable patients was estimated to be 49%. Median progression-free and overall survival was 9.5 and 21.1 months, respectively. Grade 3 and 4 hematological toxicities were neutropenia and thrombocytopenia in 44 and 5% of patients, respectively. A sustained improvement of evaluated immunological parameters with a negligible toxicity profile was observed in the 27 patients treated with interleukin 1-2/13 cis-retinoic acid. Docetaxel in combination with leucovorin, 5-fluorouracil and capecitabine followed by low-dose interleukin 1-2 and 13-cis-retinoic acid is well tolerated, and shows a significant activity in patients with metastatic gastric adenocarcinoma. PMID- 17414630 TI - Phase I/II study of irinotecan (CPT-11) and S-1 in the treatment of advanced gastric cancer. AB - A phase I/II study to determine the recommended dose for combination therapy with CPT-11 (irinotecan hydrochloride) and S-1 (tegafur, gimestat and otastat potassium) for advanced or recurrent gastric cancer, and to assess the safety and efficacy of this therapy. In the phase I portion of the study, S-1 was administered from day 1 to 14 at a fixed dose approved in Japan (80 mg/m2/day), and CPT-11 was administered on days 1 and 8, with its dose being escalated to 100 from 80 mg/m2. This regimen was repeated at 3-week intervals. The phase II portion of the study assessed the efficacy and safety of this regimen at the recommended dose determined in the phase I portion of the study. Seven patients were enrolled in the phase I portion of the study. The dose-limiting toxicity was the delay of administration owing to adverse reactions (leucopenia and diarrhea). The maximum tolerated dose of CPT-11 was 100 mg/m2 and the recommended dose was determined to be 80 mg/m2. In the phase II portion of the study, 10 patients with no prior chemotherapy regimen were enrolled. The median number of treatment cycles given was 4.5, the response rate was 20.0% (2/10) in all patients, the tumor control rate stable disease or better response was 60% (6/10) and the mean survival time was 311 days. Major adverse reactions included a decreased hemoglobin level, diarrhea, nausea and anorexia of grade 3 or worse (each occurred in 10% of the patients). Other adverse reactions were slight and well tolerated. The present combination therapy with CPT-11 and S-1 produced a low response rate but a high tumor control rate (stable disease or better response) and slight prolongation of survival time. This is a well-tolerated ambulatory regimen for advanced gastric cancer. PMID- 17414631 TI - Role of pegylated liposomal doxorubicin (Caelyx) in the treatment of relapsing ovarian cancer. AB - The most significant factor predicting response to second-line chemotherapy is the time interval elapsed from the end of chemotherapy to relapse occurrence. Two types of situations may be considered: patients with platinum-sensitive relapse (relapse-free interval longer than 6 months) and patients with platinum refractory relapse (progression during treatment or relapse-free interval under 6 months). Pegylated liposomal doxorubicin is a doxorubicin formulation. Encapsulation in liposomes confers it different pharmacokinetic characteristics and a more favorable toxicity profile. The following review examines the efficacy and safety of pegylated liposomal doxorubicin for the treatment of relapsing epithelial ovarian cancer. PMID- 17414633 TI - The challenging integration of platinum compounds, taxanes, and molecular targeted therapies in the multidisciplinary treatment of squamous cell carcinoma of the head and neck. AB - There have been important advances in the treatment of locally advanced squamous cell carcinoma of the head and neck (SCCHN). For patients with unresectable disease, the use of platinum-based chemoradiotherapy has improved the 3-year survival rate from 15-20% to 35-50%. The results of recent studies involving sequential therapy of induction chemotherapy including taxanes and chemoradiation have shown encouraging survival rates, near to 60-70%. Moreover, progress in the understanding of the molecular changes that lead to the development of these tumors is bringing novel diagnostic and therapeutic approaches into clinical practice, promising new hope to patients with a devastating and lethal disease. PMID- 17414634 TI - Chemoprevention of squamous cell carcinoma of the head and neck. AB - PURPOSE OF REVIEW: The aim of this article is to summarize progress in understanding of the biology of squamous cell carcinoma of the head and neck and of trials to prevent malignant conversion of oral premalignant lesions and the development of second primary tumors in those already treated for squamous cell carcinoma of the head and neck. RECENT FINDINGS: The understanding of squamous cell carcinoma of the head and neck biology is rapidly evolving. Clinical trials for chemoprevention are involving more diverse regimens, following disappointing results of retinoid monotherapy. In-vitro and animal studies form the rationale for the next generation of studies, employing combination, synergistic treatments. SUMMARY: Based on trial data to date, no recommendation for intervention with a chemopreventive agent can be made. It is clear, however, that smoking cessation is an effective intervention for preventing oral premalignant lesions and second primary tumors. Promising trials are being conducted and designed currently. The future of this area of study will involve rational choice of multidrug regimens based on current understanding of the biology of squamous cell carcinoma of the head and neck. PMID- 17414635 TI - Individual patients' data meta-analyses in head and neck cancer. AB - PURPOSE OF REVIEW: This is a review of the experience obtained at the Institute Gustave Roussy, evaluating the role of chemotherapy and of altered fractionated radiotherapy in locally advanced head and neck cancer. The database included nearly 120 randomized trials, and about 25,000 patients, with a median follow-up of 6 years. RECENT FINDINGS: In the chemotherapy database (Meta-Analysis of Chemotherapy in Head, Neck Cancer and Meta-Analysis of Chemotherapy in Nasopharynx Carcinoma), concomitant cisplatin-based radiotherapy-chemotherapy provided the most significant benefit on locoregional control and survival, both in head and neck squamous cell carcinomas and nasopharyngeal carcinomas. In head and neck squamous cell carcinomas, the benefit of adding concomitant chemotherapy was found to be in the same order of magnitude whether radiotherapy was postoperative or definitive. In the altered radiotherapy database (Meta-Analysis of Radiotherapy in Carcinoma of Head and Neck), among the different types of altered fractionated radiotherapy, hyperfractionation provided the most significant benefit. The benefit associated with altered fractionated radiotherapy and of concomitant chemotherapy markedly decreased with increasing age. SUMMARY: This database provided a unique tool to evaluate long-term effects of chemotherapy and altered fractionated radiotherapy in head and neck cancer. This allowed the oncological community to obtain a reliable characterization of the magnitude of the treatment benefits in this type of cancer and to base patient care and future research on strong evidence. PMID- 17414636 TI - The role of taxanes and targeted therapies in locally advanced head and neck cancer. AB - PURPOSE OF REVIEW: This review presents new data on the role of taxanes and targeted therapies in the management of squamous cell carcinoma of the head and neck. RECENT FINDINGS: Taxane-containing triplets are clearly superior as an induction regimen in locally advanced squamous cell carcinoma of the head and neck when compared with cisplatin/5 fluorouracil which has been the standard for two decades. Preliminary data suggest that the addition of a taxane to cisplatin/5 fluorouracil as induction regimen followed by chemoradiation may be superior to chemoradiation alone. The addition of cetuximab to radiation prolongs locoregional control and survival without increasing mucositis. Areas of active investigation are the search for epidermal growth factor receptor mutations and the optimal way of integrating epidermal growth factor receptor-directed therapies into standard management. Meanwhile new targets are explored. SUMMARY: Taxane/cisplatin/5 fluorouracil induction chemotherapy is clearly superior to cisplatin/5 fluorouracil. Epidermal growth factor receptor directed therapies can safely be combined with radiation and the combination shows encouraging results. PMID- 17414637 TI - Reirradiation in squamous cell head and neck cancer: recent developments and future directions. AB - PURPOSE OF REVIEW: The purpose of this review is to highlight the most important developments in the management of recurrent or second primary head and neck carcinoma in previously irradiated areas by reirradiation that have been published in the medical literature in the past year. RECENT FINDINGS: Recent research indicates that long-term survival can be achieved in a proportion of patients using more advanced chemo-reirradiation protocols in the primary as well as in postoperative reirradiation setting. Despite the promising results with regard to locoregional tumour control and survival, treatment-related acute and late morbidity remains of major concern. SUMMARY: As an increasing number of patients currently receive more effective initial treatment regimens, recurrent and second primary tumours in previously irradiated areas nowadays may represent a more radio-resistant population than reported in previous studies. Therefore, full-dose chemo-reirradiation should only be applied in well selected cases. PMID- 17414638 TI - Clinical trial design limitations in head and neck squamous cell carcinomas. AB - PURPOSE OF REVIEW: The present article reviews the randomized trials contributing to the establishment of current standards for the treatment of head and neck cancer. It provides critical analysis of their methodology in order to facilitate future trial design. RECENT FINDINGS: From a prognosis perspective, head and neck cancers are a heterogeneous group of diseases. Following a number of randomized clinical trials evaluating the role of chemotherapy in the induction, concomitant and adjuvant settings, there has been considerable improvement in the treatment of locally advanced head and neck cancers during the last decade. It is, however, difficult to interpret and compare the results optimally and to build on efficient trial designs as most of the trials included patients with different levels of essential prognostic factors. SUMMARY: All key randomized trials will be reviewed according to eligibility criteria, subgroup issues, trial power and historical controls. Methodological interpretation and possible plans for the next generation of clinical trials will be presented. PMID- 17414639 TI - Standard, and novel cytotoxic and molecular-targeted, therapies for HNSCC: an evidence-based review. AB - (1) Head and neck squamous cell carcinoma is the sixth most frequently occurring cancer worldwide.(2) Chemotherapy has shown some success as part of multimodal treatment schedules for locally advanced, nonmetastatic head and neck squamous cell carcinoma, and too a much lesser extent for metastatic head and neck squamous cell carcinoma.(3) A recent meta-analysis of 32 studies involving >10,000 patients concluded that chemotherapy added to radiotherapy produces a large survival advantage relative to radiotherapy alone.(4) Concurrent or alternate chemoradiotherapy, with a schedule based on cisplatin, has an established place in the management of locally advanced nonmetastatic head and neck squamous cell carcinoma. PMID- 17414640 TI - Testicular germ cell tumors. AB - PURPOSE OF REVIEW: Preclinical and clinical developments in germ cell tumors over the past year are summarized. RECENT FINDINGS: Attenuations in the rising incidence of testicular germ cell tumors are beginning to be observed in certain European populations. Additional data on predisposing factors related to race, estrogenic exposure, cryptorchidism, and infertility are becoming available. Significant work on the genetic and molecular alterations in tissue specimens and cell culture models of germ cell tumors continues. Additional treatment strategies for advanced stages of the disease are being evaluated. Cardiovascular and metabolic consequences of therapies in long-term testicular germ cell tumor survivors are being further clarified. Late relapses of successfully treated patients are also being increasingly recognized. SUMMARY: More effective treatments for intermediate risk, poor risk, and recurrent germ cell tumors need to be developed, while long-term toxicities of therapies need to be further modified. Given these challenges, active research on these fronts continues and remains a priority. PMID- 17414641 TI - Prostate cancer update: 2006. AB - PURPOSE OF REVIEW: This review highlights the most important developments in the diagnosis, prevention, and treatment of prostate cancer published in the medical literature over the past year. RECENT FINDINGS: Recent research has revealed a relatively high rate of prostate specific antigen screening in elderly men regardless of comorbidity status. Other findings include data that finasteride may increase the sensitivity of prostate specific antigen to detect prostate cancer. In terms of use of androgen deprivation therapy as part of primary therapy, a randomized trial has demonstrated that immediate therapy for locally advanced prostate cancer improves overall survival. There is growing literature, however, confirming the risks associated with this treatment, including an increased incidence of cardiovascular events and diabetes. New randomized trial data of adjuvant radiotherapy for high-risk disease have not demonstrated a survival benefit over observation. For patients who experience disease relapse following local therapy, definitions of biochemical failure following prostatectomy and radiation therapy are proposed. In more advanced disease, immunotherapy has preliminarily demonstrated survival benefit in a randomized trial. SUMMARY: Data reported over the last year have significant implications for those involved in the management of prostate cancer, ranging from primary care physicians to medical oncologists, urologists, and radiation oncologists. PMID- 17414642 TI - Renal cell carcinoma. AB - PURPOSE OF REVIEW: This review highlights recent contributions to the biology and treatment of renal cell carcinoma, the expanded use of antiangiogenic agents as well as interest in other inhibitory drug mechanisms. In addition, recent findings are reported on biomarkers which are undergoing investigation as correlative prognostic indicators of either survival or response to treatment. RECENT FINDINGS: Advances in our understanding of the molecular biology underpinning renal cell carcinoma and the introduction of new targeted therapeutics with benefit in the metastatic setting have had a major impact on the treatment of this disease. SUMMARY: The management of metastatic renal cell carcinoma has undergone a dramatic evolution in the past year, marked by the approval of two drugs by the US Food and Drug Administration. These drugs have demonstrated improved progression-free survival as well as potentially improved overall survival for patients with metastatic disease. These groundbreaking treatment strategies have fueled a surge in translational studies expanding our knowledge of the molecular biology of renal cell carcinoma. PMID- 17414643 TI - Bladder cancer. AB - PURPOSE OF REVIEW: This article reviews the diagnosis and management of bladder cancer with an emphasis on studies and developments over the past year. RECENT FINDINGS: Cystoscopy remains the mainstay in the detection and surveillance of bladder cancer, though efforts continue in the development of urinary bladder cancer markers. Superficial bladder cancer continues to be managed predominantly through transurethral resection with perioperative instillation of chemotherapy recommended for most patients. Intravesical bacille Calmette-Guerin (including a maintenance regimen) should be used for those at high risk for progression. Muscle invasive disease continues to be managed by radical cystectomy. Research continues on the use of laparoscopy, the effect on patient's health-related quality of life, and the potential role for bladder preservation strategies. The role of neoadjuvant versus adjuvant chemotherapy around the time of cystectomy remains to be resolved. The mainstays of chemotherapy remain methotrexate, vinblastine, doxorubicin, and cisplatin, and gemcitabine and cisplatin, but work is ongoing to develop new regimens, especially in patients who cannot take cisplatin. SUMMARY: Although great strides continue to be made each year in the diagnosis and management of bladder cancer considerably more work needs to be done in order to improve the lives of our patients with this disease. PMID- 17414644 TI - Pediatric genitourinary tumors. AB - PURPOSE OF REVIEW: We will review the 2005 and 2006 literature on pediatric genitourinary tumors. RECENT FINDINGS: Survival continues to improve for primary renal, bladder/prostate and testicular tumors in childhood. The addition of more intensive chemotherapy for anaplastic histology disease, recognition of loss of heterozygosity for chromosomes 1p and 16q as an adverse prognostic factor in favorable histology Wilms' tumor, and the utilization of molecular markers to better characterize all renal tumors will better enable individualized therapy. Recognition and treatment of anaplastic histology and bilateral Wilms' tumor remains a challenge. In rhabdomyosarcoma, genitourinary site and embryonal histology confer a relatively favorable prognosis. SUMMARY: Advances in molecular oncology, diagnostic imaging, surgical approaches and long-term follow-up of childhood cancer survivors drive risk-stratified therapy in pediatric genitourinary tumors. PMID- 17414645 TI - Bone metastases in prostate cancer: a targeted approach. AB - PURPOSE OF REVIEW: New therapies are needed for hormone refractory prostate cancer. Promising new treatments specifically target metastatic disease in the bone, the predominant site of spread in prostate cancer. Recent advances in the understanding of the biology of the tumor-bone microenvironment are leading to the development of new bone-targeted therapies. In this article, we review current and developing clinical strategies designed to control prostate-cancer bone metastases. RECENT FINDINGS: Newly developed biologic therapies that target prostate-cancer bone metastases have shown promising results in animal models and clinical trials. Additionally, currently established drugs such as bisphosphonates and bone-seeking radiopharmaceuticals are being applied in novel ways in an effort to improve patient care. SUMMARY: Therapies targeting bone in advanced prostate cancer have demonstrated improvements in morbidity, and more recently, in overall survival. The continuing development and use of these targeted therapies have the potential to alter the course of this currently fatal disease. PMID- 17414646 TI - Imaging of prostate cancer. AB - PURPOSE OF REVIEW: Appropriate imaging of prostate cancer is a crucial component of staging and therapy application. The purpose of this review is to highlight the most important developments in novel imaging modalities reported in the past year. RECENT FINDINGS: Transrectal ultrasound is used to guide needle biopsy and brachytherapy. Improved results are obtained with color and power Doppler transrectal ultrasound with sonographic contrast agents. The role of elastography in prostate cancer remains to be elucidated. Magnetic resonance imaging is now widely used for staging before treatment and accumulating data indicate the utility of this technique with magnetic resonance spectroscopy in staging and follow-up. Positron-emission tomography alone or especially in combination with computed tomography imaging with the new radiotracers (11)C-choline, (18)F fluorocholine, (11)C-acetate and (18)F-fluoride have shown promising results. Further investigations in larger clinical studies are necessary to establish the role of these imaging techniques in the management of patients with prostate cancer. SUMMARY: This report provides a summary of novel types of imaging and indicates their promise in prostate cancer. PMID- 17414647 TI - Bibliography. Current world literature. Head and neck. PMID- 17414648 TI - Presidential Address: 21st Scientific Meeting of the International Society of Hypertension: dietary sodium and cardiovascular disease: the 'J'-shaped relation. PMID- 17414649 TI - The sympathetic nervous system and the metabolic syndrome. AB - Studies performed in the past two decades have unequivocally shown that several of the components of the metabolic syndrome are associated with indirect and direct markers of adrenergic overdrive. This is the case for hypertension and obesity, in which resting tachycardia, elevated plasma norepinephrine values, increased sympathetic nerve traffic, as well as augmented levels of total and regional norepinephrine spillover have been reported. This is also the case for insulin resistance, i.e. a metabolic condition frequently complicating the various components of the pathological condition identified as the 'metabolic syndrome'. After briefly describing the epidemiological and the cardiovascular risk profile of the disease, this paper will examine the behaviour of the sympathetic nervous system in the metabolic syndrome as well as the mechanisms potentially responsible for this neurogenic abnormality. This will be followed by an analysis of the role played by neuroadrenergic factors in disease progression as well as in the pathogenesis of its complications. Finally, the therapeutic implications of these findings will be highlighted. PMID- 17414650 TI - Putting blood pressure in its place. AB - The benefit of blood pressure-lowering therapy in individuals with normal blood pressure who are at high cardiovascular risk creates a challenge in how best to advocate such therapy. Arguments based on a putative causal role for blood pressure and the need to reduce blood pressure may be less likely to succeed than arguments based on the need to prevent and to reverse the cardiovascular disease that causes cardiovascular events. PMID- 17414651 TI - What is the true place of blood pressure in cardiovascular risk management? PMID- 17414652 TI - Twenty-four-hour ambulatory blood pressure profiles of high-risk patients in general practice: data from an ambulatory blood pressure monitoring registry. PMID- 17414653 TI - Metabolic syndrome and arterial stiffness: evidence for gender disparity and early effects of non-traditional risk factors? PMID- 17414654 TI - Predicting hypertension complications from small artery structure. PMID- 17414655 TI - Left ventricular hypertrophy: beyond the image and defining the human cardiac phenotype in hypertension. PMID- 17414656 TI - Bashing diuretics or failure of surrogate endpoint? PMID- 17414657 TI - Blood pressure-dependent and independent effects of agents that inhibit the renin angiotensin system. AB - OBJECTIVES: To evaluate the blood pressure-dependent and independent effects of angiotensin-converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARB) on major cardiovascular events. METHODS: Using data from 26 large-scale trials comparing an ACEI or an ARB with placebo or another drug class, meta regression analyses were conducted in which treatment-specific relative risks for major cause-specific outcomes [stroke, major coronary heart disease (CHD) events and heart failure] were regressed against follow-up blood pressure differences. RESULTS: From a total of 146 838 individuals with high blood pressure or an elevated risk of cardiovascular disease, 22 666 major cardiovascular events were documented during follow-up. The analyses showed comparable blood pressure dependent reductions in risk with ACEI and ARB (P >or= 0.3 for all three outcomes). The analyses also showed that ACEI produced a blood pressure independent reduction in the relative risk of CHD of approximately 9% (95% confidence interval 3-14%). No similar effect was detected for ARB, and there was some evidence of a difference between ACEI and ARB in this regard (P = 0.002). For both stroke and heart failure there was no evidence of any blood pressure independent effects of either ACEI or ARB. CONCLUSION: There are similar blood pressure-dependent effects of ACEI and ARB for the risks of stroke, CHD and heart failure. For ACEI, but not ARB, there is evidence of blood pressure-independent effects on the risk of major coronary disease events. PMID- 17414658 TI - Does self-reported history of hypertension predict cardiovascular death? Comparison with blood pressure measurement in a 19-year prospective study. AB - OBJECTIVES: Hypertension as assessed by blood pressure (BP) measurement is one of the most important risk factors for cardiovascular diseases (CVD). Self-reported history of hypertension (self-reported HT) is an easy way to obtain information on BP and is known to have a certain sensitivity and high specificity for hypertension confirmed by BP measurement (confirmative HT). Thus, it might predict CVD mortality, but few studies have reported on this relationship. METHODS: We followed 6427 participants aged 30-59 years without a history of CVD for 19 years. The multivariate-adjusted hazard ratio (HR) of CVD mortality was estimated by the Cox proportional hazard model. RESULTS: The sensitivity and specificity of self-reported HT for confirmative HT were 52-65% and 95%, respectively. The multivariate-adjusted HR of self-reported HT for CVD death was 2.49 [95% confidence interval (CI) = 1.72-3.61]. Compared to participants with neither self-reported HT nor confirmative HT, those with confirmative HT showed a consistently higher HR for CVD mortality. Self-reported HT without confirmative HT was also significantly related to CVD mortality (HR = 2.10, 95% CI = 1.04 4.26). These tendencies were unchanged when we further adjusted for systolic BP (SBP) level. The age-adjusted mortality rate of individuals with self-reported HT corresponded to the age-adjusted mortality rate of individuals whose SBP was 160 179 mmHg. CONCLUSION: Self-reported HT could screen one-half of the participants for confirmative HT and was significantly associated with CVD mortality. These results indicate that self-reported HT can be a useful tool to screen for individuals with high BP if it is difficult to perform BP measurements continuously among all members of a community. PMID- 17414659 TI - Alcohol consumption and blood pressure in the adult US population: assessment of gender-related effects. AB - OBJECTIVE: Our objective was to assess the gender-related effects of alcohol consumption on blood pressure (BP) in a representative sample of the adult US population. METHODS: We examined data from the National Health and Nutrition Examination Survey 1999-2000. The effects of various risk factors for hypertension on BP were examined with analysis of covariance statistics. RESULTS: Of the 5448 adults over 20 years of age, 2650 (48.7%) reported the intake of one or more drinks per day over the past year. In this population, the mean +/- SEM age was 46.9 +/- 0.34 years, the body mass index was 24.8 kg/m, 1257 (47.4%) were women, systolic BP was 124.3 +/- 0.44 mmHg and diastolic BP was 72.7 +/- 0.27 mmHg. Hypertension was reported in 21.1%, diabetes in 5.1% and cigarette smoking in 39.7%. A significant effect on systolic BP was shown with age (P < 0.01), body mass index (P < 0.01), race (P = 0.01), gender (P < 0.01) and diabetes (P < 0.01). The interaction with gender and alcohol drinking level was significant (P = 0.02). Post-hoc analysis localized the source of this effect. There was a significant increase in systolic BP between one and three and between one and four, but not between one and two, drinks per day in men. This effect was not observed in women. CONCLUSION: Consistent with previous reports, our study suggests that alcohol intake up to two drinks per day has no effect on BP. There was a gender-related effect of alcohol intake in excess of two drinks per day on BP, with increased BP observed only in men but not in women. PMID- 17414660 TI - Age-dependent association of alcohol drinking with pulse pressure. AB - BACKGROUND: Wide pulse pressure is a potent risk factor for atherosclerotic disease, and hypertension is induced by habitual alcohol drinking. This concise study investigated whether age affects the relationship between alcohol drinking and pulse pressure. METHODS: The participants were 99 661 male workers (20-69 years old) who had received periodic health check-ups. The mean levels of pulse pressure and blood pressure were compared among three different groups divided by the average amount of daily alcohol intake (nondrinkers; light drinkers, < 30 g ethanol/day; heavy drinkers, >or= 30 g ethanol/day) in each age group. RESULTS: The mean pulse pressure was decreased in the thirties and forties age groups compared with that in the twenties age group, and then was increased with advance of age in the fifties and sixties age groups. Pulse pressure was significantly wider in heavy drinkers than in nondrinkers and light drinkers in all age groups, and the difference between pulse pressure of nondrinkers and that of heavy drinkers was greater in the fifties and sixties age groups than in the younger groups. Pulse pressure in light drinkers was significantly wider than that in nondrinkers in the fifties and sixties age groups but was not different from that in nondrinkers in the twenties to forties age groups. CONCLUSIONS: Association of alcohol intake with pulse pressure is dependent on the amount of alcohol and is stronger in the elderly. Age as well as amount of alcohol intake should therefore be taken into account when the influence of alcohol drinking on pulse pressure is considered. PMID- 17414661 TI - Ambulatory blood pressure monitoring in hypertensive patients with high cardiovascular risk: a cross-sectional analysis of a 20,000-patient database in Spain. AB - OBJECTIVE: To evaluate ambulatory blood pressure monitoring (ABPM) parameters in a broad sample of high-risk hypertensive patients. METHODS: The Spanish Society of Hypertension is developing a nationwide project in which more than 900 physicians send ABPM registries and corresponding clinical records to a central database via www.cardiorisc.com. Between June 2004 and July 2005 a 20 000-patient database was obtained; 17 219 were valid for analysis. RESULTS: We identified 6534 patients with high cardiovascular risk according to the 2003 European Society of Hypertension/European Society of Cardiology guidelines stratification score. Office blood pressure (BP) was 158.8/89.9 mmHg and 24-h BP was 135.8/77.0 mmHg. Patients with grade 3 BP in the office showed ambulatory systolic BP values less than 160 mmHg in more than 80%. A non-dipping pattern was observed in 3836 cases (58.7%), whereas this abnormality was present in 47.9% of patients with low to-moderate risk [odds ratio (OR) 1.54; 95% confidence interval (CI) 1.45-1.64]. The prevalence of non-dippers was higher as ambulatory BP increased ( approximately 70% when 24-h systolic BP > 155 mmHg) and was similar in both groups. At the lowest levels of BP (24-h systolic BP < 135 mmHg) a non-dipping pattern was more prevalent in high-risk cases (56.6 versus 45.7%; OR 1.51; 95% CI 1.40-1.64). CONCLUSION: There was a remarkable discrepancy between office and ambulatory BP in high-risk hypertensive patients. The prevalence of a non-dipper BP pattern was almost 60%. In the lowest levels of ambulatory BP, high-risk patients showed a higher prevalence of non-dipping BP than lower-risk cases. These observations support the recommendation of a wider use of ABPM in high-risk hypertensive patients. PMID- 17414662 TI - Heritability of left ventricular mass in Japanese families living in Hawaii: the SAPPHIRe Study. AB - OBJECTIVE: Established determinants of left ventricular (LV) mass explain only a modest fraction of its variability. Family studies to date suggest that a proportion of the unexplained variability can be accounted for by additive polygenic effects. An estimate of this proportion has not been reported previously in an East Asian population. The objective of this study was to estimate the heritability of LV mass in Japanese families living in Hawaii. DESIGN AND METHODS: We analyzed data by components of variance in a sample of 169 hypertensive families (n = 476 subjects) and, separately, in a population-based sample of 256 families (n = 501 subjects) participating in the Honolulu Heart Program. RESULTS: In multivariate models, established predictors of LV mass explained about half the total variance of LV mass. Using SOLAR, our estimates of the narrow sense heritability of LV mass ranged from 42.5% (SE 9.8, P < 0.0001) in our sample of hypertensive families to 60.6% (SE 11.7, P < 0.0001) in our population-based sample of families. Parametric bootstrap analyses confirmed that the inference for each sample was appropriate. CONCLUSIONS: Assuming the absence of shared familial environmental effects, close to half of the unexplained variance of LV mass in Japanese subjects living in Hawaii is genetic in nature. This estimate was observed in two independent samples. Therefore, the pursuit of novel genetic determinants of LV mass through either whole genome or candidate gene association studies of this population may be worthwhile. Such studies are certainly feasible. PMID- 17414663 TI - Hypertension and hypothyroidism: results from an ambulatory blood pressure monitoring study. AB - OBJECTIVE: To examine differences between hypothyroid patients and healthy volunteers in 24-h ambulatory blood pressure parameters. METHODS: The study population consisted of 100 individuals who were recently diagnosed for hypothyroidism. These patients had never been treated before with antihypertensive treatment or received drugs for hypothyroidism. All participants underwent 24-h ambulatory blood pressure monitoring. The control group consisted of 100 healthy volunteers matched one to one for gender and age with the hypothyroid participants. RESULTS: Clinic systolic and diastolic blood pressures were significantly higher in patients with hypothyroidism compared with volunteers. The mean 24-h systolic blood pressure and 24-h pulse pressure were significantly higher in patients with hypothyroidism compared with volunteers. The 24-h systolic blood pressure variability was also significantly higher in patients with hypothyroidism. Fasting serum cholesterol tended to be higher in patients with hypothyroidism compared with volunteers but the difference was not statistically significant, while fasting serum triglycerides were significantly higher. Body mass index was also significantly higher in patients with hypothyroidism. CONCLUSIONS: These findings indicate that hypothyroidism may be an important predictor of higher mean 24-h systolic blood pressure, 24-h pulse pressure and 24-h systolic blood pressure variability, parameters of ambulatory blood pressure monitoring that have been previously associated with higher cardiovascular target organ damage. PMID- 17414664 TI - Evidence for an estradiol-agonistic action of nebivolol in spontaneously hypertensive rats. AB - OBJECTIVES: Unlike classical beta1-selective blockers, nebivolol (NEB) has vasodilatory properties due to the release of nitric oxide (NO) by a mechanism that is, so far, unknown. We hypothesized that NEB stimulates NO release by binding to estrogen receptors (ER) and subsequent activation of endothelial NO synthase (eNOS). The aim of this study was to elucidate the underlying mechanism of NEB action by investigating estradiol-dependent effects of NEB on the NO system in spontaneously hypertensive rats (SHR). METHODS: The effects of NEB on the NO system were determined by measuring urinary nitrate/nitrite (NOx) as well as eNOS and caveolin-1 protein expression in aortae. RESULTS: NEB did not influence NOx excretion in sham-operated (SO) female rats during proestrus. In male and ovariectomized female (OVX) rats, NEB increased NOx excretion significantly, whereas N(G)-nitro-L-arginine methyl ester (L-NAME) inhibited the NEB-induced increase in NOx. ER blockade with ICI182,780 prevented NEB-induced NOx excretion in OVX rats. In the aortae of SO females, NEB treatment did not alter eNOS expression. In OVX rats eNOS expression was increased two-fold after NEB application and this could be prevented by pretreatment with ICI182,780. In contrast to eNOS, NEB did not influence caveolin-1 expression in either group. CONCLUSION: The ability of NEB to up-regulate NOx excretion in male and OVX SHR and the inhibitory effect of ICI182,780 on NEB-induced NOx excretion suggests that NEB has an estradiol-agonistic action in vivo. NEB provokes NO generation by up-regulation of eNOS protein expression, whereas the expression of the negative eNOS regulator caveolin-1 remains unaffected. PMID- 17414665 TI - Clustering of metabolic syndrome risk factors and arterial stiffness in young adults: the Northern Ireland Young Hearts Project. AB - OBJECTIVES: This study aimed to investigate whether the clustering of the risk factors of the metabolic syndrome (MetS) is associated with stiffness of central and peripheral arterial segments; whether these associations are similar in men and women; and whether insulin resistance and low-grade inflammation mediate any such associations. BACKGROUND: Increased arterial stiffness may explain, at least in part, the increased cardiovascular and diabetes risk associated with the MetS. However, the mechanisms linking the MetS to an increased arterial stiffness are incompletely understood, and gender differences may exist. METHODS: Cross sectional analyses of data on 313 young men and women (mean age 23 years) from the Northern Ireland Young Hearts Project. Subjects were categorized according to the number of traits of the MetS; in addition a continuous MetS score was calculated. Arterial stiffness was assessed by measuring pulse wave velocity (PWV) in three arterial segments using a non-invasive optical method. RESULTS: The prevalence of the MetS was similar for men (10.6%) and women (10.5%). After adjustment for potential confounders and other cardiovascular risk factors, PWV of the three arterial segments investigated increased with increasing traits of the MetS in women only. Women with the MetS, as compared to those without risk factors of the syndrome, had greater PWV of the aorto-iliac (+14.0%, P = 0.016), the aorto-radial (+13.2%, P = 0.010) and aorto-dorsalis pedis (+11.8%, P = 0.011) segments. A great deal (up to 75%) of the association between the MetS and aortic iliac PWV was mediated by heart rate, inflammation markers [C-reactive protein (CRP) and fibrinogen] and insulin resistance [homeostatic model assessment insulin resistance (HOMA-IR)], whereas these variables did not explain much of the association between the MetS and PWV of the peripheral segments. CONCLUSIONS: Young women with the MetS show increased stiffness of peripheral and central arteries, a mechanism that may explain their increased cardiovascular risk. Low grade inflammation, insulin resistance and sympathetic activation explain much of the adverse impact of the MetS on central, but not peripheral, arterial stiffness. PMID- 17414666 TI - Small artery structure is an independent predictor of cardiovascular events in essential hypertension. AB - OBJECTIVE: Structural abnormality of resistance arteries is a characteristic pathophysiological phenomenon in essential hypertension and can be assessed in vitro as an increase in the media: lumen ratio (M: L) of isolated small arteries. We have investigated whether M: L is a risk predictor in uncomplicated essential hypertensive patients. Recently, high M: L was demonstrated as a prognostic marker in patients at high cardiovascular risk, including normotensive type 2 diabetic patients. Since diabetes is associated with pressure-independent changes in M: L, the relevance of this finding to essential hypertension has been uncertain. METHODS: We conducted a follow-up survey of 159 essential hypertensive patients, who had previously been submitted to a M: L evaluation while participating in a clinical trial. They composed a homogeneous moderate-risk group, with no concomitant diseases, and represented 1661 years of follow-up. RESULTS: Thirty patients suffered a documented predefined cardiovascular event during follow-up. Increased relative risk (RR) was associated with M: L >or= 0.083 (mean level of the hypertensive cohort), RR = 2.34 [95% confidence interval (CI) 1.11-4.95], and with M: L >or= 0.098 (mean level of a normotensive control group + 2SD), RR = 2.49 (95% CI 1.21-5.11). Both results remained significant (RR = 2.19, 95% CI 1.04-4.64, and RR = 2.20, 95% CI 1.06-4.56, respectively) when adjusted for Heart Score level (10-year mortality risk-estimate, integrating age, gender, systolic blood pressure, cholesterol and smoking). CONCLUSION: Abnormal resistance artery structure independently predicts cardiovascular events in essential hypertensive patients at moderate risk. PMID- 17414667 TI - Small artery structure adapts to vasodilatation rather than to blood pressure during antihypertensive treatment. AB - OBJECTIVE: Correction of the abnormal structure of resistance arteries in essential hypertension may be an important treatment goal in addition to blood pressure (BP) reduction. We investigated how this may be achieved in a prospective clinical study. METHODS: Plethysmography was used to measure forearm resting vascular resistance (Rrest) and minimum vascular resistance (Rmin) as a measure of vascular structure. Two different groups of patients with essential hypertension were examined at baseline and after 6 months of antihypertensive treatment. In group A, 21 patients with never-treated essential hypertension were treated by their general practitioners using a variety of drugs to allow an assessment of the drug-independent effects. In group B, 28 beta-blocker-treated patients were shifted to angiotensin II receptor blocker treatment (eprosartan) to allow vasodilatation with no change in BP. RESULTS: In group A, mean ambulatory blood pressure (ABP) fell from 119 +/- 2 (SE) to 103 +/- 2 mmHg (P < 0.01), whereas mean ABP was unchanged in group B (100 +/- 1 to 99 +/- 1 mmHg, P = NS). Both groups showed similar reductions in Rrest (-33.4 and -28.5%, respectively) and in Rmin (-15.4 and -15.6%, respectively). There was a strong correlation between changes in Rrest and Rmin within both groups (r = 0.57, P < 0.01 and r = 0.68, P < 0.0001, respectively), whereas the change in BP in group A was not correlated to the change in Rmin (r = -0.03). CONCLUSION: The correction of forearm resistance artery structure during antihypertensive treatment depends on the vasodilatation achieved rather than BP reduction. PMID- 17414668 TI - Arterial stiffness as an independent predictor of longitudinal changes in cognitive function in the older individual. AB - BACKGROUND: Cognitive decline significantly contributes to disability in older individuals. We previously demonstrated cross-sectionally that arterial stiffness [pulse wave velocity (PWV)] was associated with memory loss independently of traditional cardiovascular risk factors and of neuroimaging findings in older individuals without prior stroke. The present study aimed to evaluate PWV as a predictor of longitudinal changes in cognitive function in older individuals reporting memory problems. PARTICIPANTS AND METHODS: We studied 102 older individuals (mean age 79 +/- 6 years; 31 men, 71 women) reporting memory problems. PWV was measured noninvasively by Complior. Traditional cardiovascular risk factor levels were measured. Global cognitive function was measured by the Mini-Mental State Examination (MMSE) (maximum score = 30) at baseline and at follow-up visit. Cerebral computed tomography evaluated the presence of microvascular damage or cortical atrophy. Individuals with prior stroke or atrial fibrillation were excluded. RESULTS: The baseline MMSE was 22.9 +/- 5.5; 61% were hypertensive, 26.8% diabetic, 9.4% smokers, 10.5% taking statins, and 21.1% taking nitrates. The average PWV was 13.5 +/- 2.2 m/s. After a median follow-up of 12 months, the average per-year decline in MMSE was 2.9 points or 12.1%. Multiple regression models showed that PWV independently predicted cognitive decline (model R2 = 0.50). PWV was the single strongest predictor of cognitive decline, explaining 15.2% of the total variance (each 1 m/s increase in PWV was associated with an average 0.74 per-year decrease in MMSE score, P < 0.001). CONCLUSION: In older individuals, arterial stiffness (PWV) is a strong predictor of loss in cognitive function, independent of age, sex, education, and traditional cardiovascular risk factors. PMID- 17414669 TI - Enhanced angiotensin II production by renal mesangium is responsible for apoptosis/proliferation of endothelial and epithelial cells in a model of malignant hypertension. AB - OBJECTIVE: The systemic renin-angiotensin system (RAS) plays a crucial role in the pathogenesis of malignant hypertension. However, the intrarenal RAS might be at least equally important. We investigated the relationship between intrarenal RAS and mesangial, epithelial and endothelial cell proliferation/apoptosis in a model of malignant hypertension. METHODS: Cultured murine mesangial cells were subjected to 160 mmHg hydrostatic pressure for 1 h. Angiotensin II was assessed by radio-immunoassay (RIA); pro-metalloproteinase-1 (pro-MMP-1) by enzyme-linked immunosorbent assay (ELISA); hydrogen peroxide (H2O2) by photocolorimetric assay, apoptosis by terminal dUTP (2-deoxyuridine 5'-triphosphate) nick-end labelling (TUNEL), p53 by western blot and proliferation by [H]thymidine incorporation, with or without angiotensin II and/or angiotensin II type 1/angiotensin II type 2 (AT-1/AT-2) receptor blockers. Endothelial and epithelial cells were similarly treated, and the same parameters evaluated. Further, untreated cells of both lines were cultured in conditioned medium of mesangial cells exposed to pressure. Their proliferation, apoptosis and angiotensin II production were also assessed. RESULTS: High hydrostatic pressure increased angiotensin II production by mesangial cells, coinciding with augmented apoptosis and proliferation. Co stimulation with exogenous angiotensin II amplified both effects. Pressure per se evoked no response in endothelial/epithelial cells, while exogenous angiotensin II stimulated proliferation and apoptosis. No augmentation of p53 expression was evident. These effects were abolished by anti-angiotensin-II peptide, saralasine and losartan, but not by PD123319. Incubation of untreated cells in medium of mesangium subjected to pressure, augmented proliferation and apoptosis. No significant changes were noticed in pro-MMP or H2O2. CONCLUSIONS: Mesangium plays a deleterious role in the pathogenesis of malignant hypertension. High hydrostatic pressure stimulates angiotensin II synthesis by mesangial cells. The latter is responsible for hypercellularity and apoptotic death of mesangial, endothelial and epithelial cells. In this model, exaggerated apoptosis and proliferation are mediated via the angiotensin II pathway independently of p53 gene activation. PMID- 17414670 TI - Inhibition of Rho-kinase attenuates nephrosclerosis and improves survival in salt loaded spontaneously hypertensive stroke-prone rats. AB - OBJECTIVES: We examined whether the Rho/Rho-kinase pathway is involved in the pathogenesis of nephrosclerosis in severely hypertensive rats and assessed the effects of long-term treatment with a Rho-kinase inhibitor, fasudil, on kidney function, histological findings, gene expressions, and survival. We also attempted to elucidate the mechanisms involved. METHODS: We studied the following four groups: control Wistar-Kyoto rats (WKY), untreated salt-loaded spontaneously hypertensive stroke-prone rats (SHR-SP), low-dose fasudil (15 mg/kg per day) treated SHR-SP, and high-dose fasudil (30 mg/kg per day)-treated SHR-SP. After 8 weeks' treatment, the effects of fasudil were examined. RESULTS: Untreated SHR-SP were characterized by increased blood pressure without circadian variation, decreased kidney function, abnormal renal morphological findings, and increased messenger RNA expression levels of transforming growth factor beta, collagen I, collagen III, p40phox, p47phox, plasminogen activator inhibitor 1, and intracellular adhesion molecule 1 in the renal cortex, compared with WKY. Long term high-dose fasudil treatment significantly improved renal function (serum creatinine -32%, creatine clearance +39%), proteinuria (-92%) and histological findings (glomerular injury score -57%, arteriolar injury score -55%, fibrous area -40%, ED-1-positive cells -43%) without changing blood pressure or circadian variation, compared with untreated SHR-SP. In addition, fasudil significantly improved increased mRNA expression levels in the renal cortex. Furthermore, high dose fasudil significantly prolonged survival time compared with untreated SHR-SP (P < 0.01). Low-dose fasudil treatment improved these variables slightly, but did not affect most significantly. CONCLUSION: The Rho/Rho-kinase pathway participates in the pathogenesis of nephrosclerosis in SHR-SP independently of blood pressure-lowering activity, partly by upregulation of the gene expressions of extracellular matrix, oxidative stress, adhesion molecules, and antifibrinolysis. PMID- 17414671 TI - Function and regulation of epithelial sodium transporters in the kidney of a salt sensitive hypertensive rat model. AB - OBJECTIVE: To determine the function and regulation of thiazide-sensitive NaCl co transporters (NCC), NaK2Cl co-transporters (NKCC2), and epithelial sodium channels (ENaC) in the kidneys of a salt-sensitive hypertensive model. DESIGN AND METHODS: Neonatal Wistar rats were treated with capsaicin or vehicle. Seven-week old male rats were treated for 2 weeks with: vehicle plus a normal (Con-NS) or high (Con-HS) sodium diet, and capsaicin pretreatment plus a normal (Cap-NS) or high (Cap-HS) sodium diet. Mean arterial pressure (MAP), renal excretory function, and protein expression determined by western blot were performed. RESULTS: MAP was increased in Cap-HS compared with other groups. Trichlormethiazide increased urine sodium excretion (UNaV) and urine flow rate (UFR) and decreased MAP in Cap-HS rats only. Furosemide increased UNaV and UFR in Cap-NS, Con-HS and Cap-HS, and decreased MAP in Cap-HS rats only. Amiloride had no effect on UNaV, UFR and MAP in any group. Renal NCC contents were increased in Cap-HS compared with Con-NS, Con-HS and Cap-NS rats, and NKCC2 expression was increased in Cap-NS, Con-HS and Cap-HS compared with Con-NS rats. No change was found in ENaC alpha subunit expression. The capsaicin-induced release of calcitonin gene-related peptide from renal tissues was decreased in Cap-HS and Cap-NS compared with Con-HS and Con-NS rats. CONCLUSION: NCC and possibly NKCC2, but not ENaC, were functionally upregulated in the kidneys of rats subjected to sensory nerve degeneration plus high salt intake, suggesting that sensory neurotransmitters may regulate the expression of the former but not the latter, which may underlie the development of salt-sensitive hypertension in this model. PMID- 17414672 TI - Excessive increase in left ventricular mass identifies hypertensive subjects with clustered geometric and functional abnormalities. AB - BACKGROUND: Left ventricular mass (LVM) exceeding needs to sustain haemodynamic load has been termed 'inappropriate left ventricular mass'. We hypothesized that inappropriate LVM identifies hypertensive patients with clustered cardiac geometric and functional abnormalities. METHODS: For this purpose, 359 hypertensive individuals without prevalent cardiovascular disease underwent Doppler echocardiography. Observed LVM exceeding more than 28% of the value predicted for individual cardiac work, body size and sex was defined as inappropriate LVM. Concentric left ventricular geometry was defined as age adjusted relative wall thickness (RWT) greater than 0.40. Systolic dysfunction was defined as ejection fraction less than 50% or midwall shortening less than 14.7%. Diastolic dysfunction was defined as isovolumic relaxation time (IVRT) greater than 100 ms, E-velocity deceleration time greater than 220 ms or age and heart rate-normalized early/late (E/A) ratio less than 0.66. Left ventricular hypertrophy (LVH) was defined as an LVM index greater than 49.2 g/m2.7 in men and 46.7 g/m2.7 in women. RESULTS: As expected, inappropriate LVM was associated with higher RWT, lower left ventricular systolic function, longer IVRT and prolonged E deceleration time (all P < 0.05). Patients with inappropriate LVM had a higher prevalence of concentric geometry (65.5 versus 40.4%), systolic dysfunction (67.9 versus 47.4%) and diastolic dysfunction (46.4 versus 39%; all P < 0.001) than those with LVH. Inappropriate LVM had greater sensitivity (0.89 versus 0.54) and specificity (0.82 versus 0.62; both P < 0.01) than LVH in identifying patients with clustered left ventricular concentric geometry, systolic and diastolic dysfunction. CONCLUSIONS: Inappropriate LVM is associated with a cluster of concentric left ventricular geometry, delayed left ventricular relaxation and reduced systolic performance. Compared with LVH, inappropriate LVM is more accurate at identifying patients with clustered left ventricular geometric and functional abnormalities. PMID- 17414673 TI - Electrocardiographic characteristics and metabolic risk factors associated with inappropriately high left ventricular mass in patients with electrocardiographic left ventricular hypertrophy: the LIFE Study. AB - BACKGROUND: To investigate electrocardiographic (ECG) and metabolic abnormalities associated with left ventricular (LV) mass inappropriately high for workload and body size (termed 'inappropriate left ventricular mass'; ILVM) in hypertensive patients with ECG left ventricular hypertrophy (LVH). METHODS: In patients enrolled in the Losartan Intervention for Endpoint Reduction (LIFE) Echocardiographic Substudy, LV structure and functions were assessed by echocardiography; Sokolow-Lyon and Cornell voltage, QRS duration, Cornell voltage duration product and ST strain pattern in leads V5-V6 were evaluated on standard ECG tracings. ILVM was defined as observed LV mass greater than 128% of that predicted by sex, body size and stroke work. RESULTS: In univariate analysis, compared with subjects with appropriate LV mass (n = 593), ILVM (n = 348) was associated with older age, diabetes, higher body mass index, lower systolic blood pressure, higher serum creatinine and urinary albumin/creatinine levels, higher LV mass index and greater prevalence of wall motion abnormalities (all P < 0.05). ILVM was associated with higher Cornell voltage and voltage-duration product but not higher Sokolow-Lyon voltage, with longer QRS and higher prevalences of ECG ST strain and echocardiographic wall motion abnormalities, independent of covariates including echocardiographically defined LVH or LV geometry. In separate logistic models, the likelihood of ILVM was significantly related to prolonged QRS duration, higher Cornell voltage, and greater Cornell voltage-duration independently (all P < 0.01). CONCLUSION: In hypertensive patients with ECG LVH, ILVM was associated with prolonged QRS duration and higher Cornell voltage, with ECG ST strain pattern, and with echocardiographic wall motion abnormalities independent of traditionally defined LVH. PMID- 17414674 TI - Assessment of long-term antihypertensive treatment by clinic and ambulatory blood pressure: data from the European Lacidipine Study on Atherosclerosis. AB - OBJECTIVES: Information on the features of long-term modifications of clinic and 24-h ambulatory blood pressure (ABP) by treatment is limited. The present study aimed to address this issue. METHODS: Ambulatory BP monitoring and clinic BP (CBP) measurements were performed at baseline and at yearly intervals over a 4 year follow-up period in 1523 hypertensives (56.1 +/- 7.6 years) randomized to treatment with lacidipine or atenolol in the European Lacidipine Study on Atherosclerosis (ELSA). RESULTS: CBP was always greater than ABP, while reductions in all BP values (greater for CBP than for ABP) were on average maintained throughout 4 years, CBP changes showing limited relationship with ABP changes (r = 0.14-0.27). BP reductions by treatment during daytime and night-time were correlated (r = 0.63-0.73). BP normalization was achieved in a greater percentage of patients for CBP (41.7%) than for ABP (25.3%), with systolic BP control being always less common than diastolic BP control. BP normalization was more frequent at single yearly visits than throughout the 4 years. Twenty-four hour BP variability was reduced by treatment over 4 years in absolute but not in normalized units. CONCLUSIONS: The present study provides the best evidence available on long-term effect of antihypertensive treatment on both ABP and CBP. On average, ABP was sustainedly reduced by treatment throughout the follow-up period, but 24-h BP was more difficult to control than CBP. In several patients, ABP control was unstable between visits, the percentage of patients under control over 4 years being much less than that of those controlled at each year. Treatment induced a reduction in absolute but not in normalized BP variability estimates. This has clinical implications because of the prognostic importance of ABP mean values and variability. PMID- 17414675 TI - Superior effect of an angiotensin-converting enzyme inhibitor over a diuretic for reducing aortic systolic pressure. AB - BACKGROUND: In recent studies, benefit has been shown for angiotensin-converting enzyme (ACE) inhibitors and calcium antagonists over a beta-blocker in hypertension, through a greater reduction in aortic than brachial systolic and pulse pressure. No data are available on diuretics, even though these are the preferred initial treatment of patients with mild hypertension. METHODS: In this study, 101 patients with mild essential hypertension were randomly assigned to an 8-week period of monotherapy with enalapril 10 mg a day or indapamide 2.5 mg a day. Central as well as brachial systolic, augmented, and pulse pressure were determined using SphygmoCor, as in the REASON and CAFE trials. RESULTS: Enalapril and indapamide reduced brachial systolic, diastolic, mean and pulse pressure to the same extent, and the heart rate was unchanged. Although there was no difference in brachial pressure with enalapril and indapamide, enalapril caused a greater fall in estimated aortic systolic, and pulse pressures. The augmentation index, an index of wave reflection, fell from 33.7 to 28.3% with enalapril but was unchanged with indapamide. CONCLUSION: Results infer a reduction in wave reflection with enalapril, causing a fall in aortic pressure augmentation, and a corresponding fall in aortic systolic and pulse pressure. These were not apparent from brachial cuff measurements. Results show that a diuretic, like a beta blocker agent, is not as effective a therapy as an ACE inhibitor in reducing aortic systolic and pulse pressure, and that the difference is not attributable to a change in heart rate. PMID- 17414677 TI - ESH update on hypertension management. PMID- 17414678 TI - Usefulness of the vasodilator minoxidil in resistant hypertension. PMID- 17414681 TI - Is anorexia nervosa a delusional disorder? An assessment of eating beliefs in anorexia nervosa. AB - Anorexia nervosa (AN) is a serious mental illness, characterized in part by intense and irrational beliefs about shape and weight, including fear of gaining weight. Although these beliefs are considered to be a diagnostic criterion for the illness, they have not been systematically characterized. This study used the Brown Assessment of Beliefs Scale (BABS) to identify the dominant belief that interfered with eating in a sample of underweight patients with AN (N=25). The degree of insight was assessed quantitatively. The majority of participants (68%) spontaneously reported a dominant belief consistent with fear of gaining weight or becoming fat. Twenty percent of patients were categorized as delusional. The total score on the BABS was significantly correlated with the drive-for-thinness subscale of the Eating Disorders Inventory (EDI) (r=0.41, p=0.04), but did not correlate with overall measures of AN severity (body mass index [BMI], duration of illness, lowest BMI, other subscales of the EDI, or total EDI score). These findings highlight the centrality of fear of fat in AN and suggest the possibility that there is a subgroup of patients whose concerns about their weight reaches delusional proportions. This subpopulation of patients warrants further study, since patients with more delusional beliefs may have a form of AN that is more refractory to treatment. PMID- 17414682 TI - Pharmacologic reduction of CNS noradrenergic activity in PTSD: the case for clonidine and prazosin. AB - This article reviews the neurobiologic rationale for and presents clinical guidance concerning the use of medications that reduce central nervous system noradrenergic activity in the treatment of intrusive symptoms of posttraumatic stress disorder. The authors reviewed neurobiological studies, nonclinical studies using animal models, clinical case reports, open-label drug studies, and blinded, placebo-controlled drug studies. This review of the basic science and clinical literature, and the authors' clinical experience with culturally and demographically diverse populations, indicate that clonidine and prazosin can play a useful role in treating sleep disturbance and hyperarousal in posttraumatic stress disorder, with minimal adverse effects and low financial cost. PMID- 17414683 TI - Medication adherence and the use of long-acting antipsychotics in bipolar disorder. AB - BACKGROUND: Adherence to treatment is a major determinant of outcome in bipolar disorder. Poor insight, attitudes towards treatment, and poor understanding of medications and the illness can all lead to reduced adherence. Nonadherence and partial adherence both also appear to play a significant role in relapse. Thirty to forty percent of patients with bipolar disorder who attempt to be adherent to treatment are actually only partially adherent. Clinicians frequently address the problem of poor adherence by adding an antipsychotic medication to the mood stabilizer regimen. The availability of a long-acting atypical antipsychotic raises the possibility of using this agent to prevent bipolar relapse. METHODS: The literature on the use of depot antipsychotics in bipolar illness is reviewed, based on a search of PubMed and Ovid Medline. RESULTS: No randomized, controlled trials of depot antipsychotics in bipolar illness have been performed. However, several case series and naturalistic trials that have used first generation agents suggest that depot antipsychotics are effective in reducing relapse in bipolar illness. CONCLUSIONS: Depot antipsychotics, including long-acting first and second generation agents, can be important adjuncts in the long-term management of bipolar illness. Controlled trials with these agents in bipolar disorder are warranted. PMID- 17414684 TI - An open-label study of olanzapine in children and adolescents with schizophrenia. AB - The purpose of this open-label study was to evaluate the use of olanzapine in the treatment of children and adolescents with schizophrenia. Sixteen children who were 8-17 years of age and met DSM-IV criteria for schizophrenia were admitted into a 10-week, open-label, optimizing dose study of olanzapine. The Brief Psychiatric Rating Scale (BPRS), the Positive and Negative Syndrome Scale (PANSS), and the Clinical Global Impression (CGI)-Severity/Improvement scales were used to assess improvement during the study. Of the 16 subjects who completed the study, 12 demonstrated significant improvement on end of treatment BPRS, CGI, and PANSS scores compared with baseline. Male subjects showed greater improvement and also gained more weight. Weight gain occurred in all but 2 subjects. Weight gain was significant, with patients averaging a gain of about 6.2 kg during the 6-week course of the study. Two of the subjects experienced extrapyramidal symptoms. The average dose of olanzapine for all subjects was 0.17 mg/kg. PMID- 17414685 TI - Polypharmacy: when is it rational? AB - In this article, the authors discuss when it makes sense to consider using more than one medication to treat a single condition. They give a brief history of the use of polypharmacy in psychiatry and discuss how new discoveries in psychotropic drug development are making polypharmacy an increasingly important topic today. The authors then present a list of 10 criteria to guide the rational use of psychotropic polypharmacy and explain each in detail with examples drawn from clinical practice. PMID- 17414686 TI - Deconstructing depression. PMID- 17414687 TI - Issues related to possession of firearms by individuals with mental illness: an overview using California as an example. PMID- 17414688 TI - PsychiatryOnline. PMID- 17414689 TI - Neuroleptic malignant syndrome induced by low dose aripiprazole in first episode psychosis. PMID- 17414690 TI - Successful monotherapy treatment with aripiprazole in a patient with schizophrenia and prolactinoma. PMID- 17414691 TI - Residual anxiety symptoms in depressed primary care patients. AB - BACKGROUND: The goal of this study was to characterize the burden of anxiety among residual depressive symptoms in naturalistic primary care settings. METHODS: A post-hoc analysis of a database comprised of naturalistically treated depressed patients across Canada was done. This bilingual (English and French), multi-center, randomized validation study was conducted in 47 primary care settings in four provinces of Canada. Patients who met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text-Revision (DSM-IV-TR) criteria for a major depressive episode, in the context of a major depressive disorder (N=454) were enrolled. Eligible patients received open-label, flexible-dose antidepressant treatment. The analysis reported here was limited to patients whose depression severity was evaluated using the Hamilton Depression Rating Scale (HAMD-17) (n=205). Patients completing 8 weeks of open-label antidepressant treatment (n=157) were considered evaluable. As a proxy for anxiety symptoms, scores on 6 items from the HAMD-17 (psychological anxiety, somatic anxiety, gastrointestinal distress, fatigue, hypochondriasis, and insight into illness) were summed to arrive at a composite anxiety score, which was then used to calculate an anxiety ratio (with the composite anxiety score as the numerator and the total HAMD-17 score as the denominator). RESULTS: The composite anxiety ratio at baseline did not correlate with the probability of remitting at endpoint (p=0.534). After 8 weeks of antidepressant therapy, remitting patients evinced a statistically significant decrease in anxiety ratio (p=0.041). Moreover, an inverse correlation was noted between severity of anxious symptoms at endpoint and probability of remission (p=0.026). The burden of anxiety, presented as the anxiety ratio, was higher in non-remitting patients at endpoint (p=0.828). CONCLUSION: Residual depressive symptoms represent ongoing illness activity in depression. Sharpening the focus of therapeutic interventions in the clinical environment calls for tracking and managing residual anxiety symptoms. PMID- 17414692 TI - A study of polypharmacy with second generation antipsychotics in patients with severe and persistent mental illness. AB - BACKGROUND: There is a paucity of empirical support for polypharmacy with second generation (atypical) antipsychotics (SGAs), especially in understudied populations. OBJECTIVE: To investigate the frequency, effectiveness, and safety of this practice in patients with severe and persistent mental illness who are chronically hospitalized. METHODS: A chart review was conducted at a state psychiatric hospital in Syracuse, NY. The study subjects (N=26) were chronically hospitalized individuals with DSM-IV diagnoses of schizophrenia or schizoaffective disorder who were initially prescribed at least one SGA and then received at least one other SGA during the study period. Demographic and clinical data were collected. Baseline and 6-month assessments were compared for statistical significance (p<0.05). RESULTS: Of the 117 chronically hospitalized inpatients at the study center, 22.2% (N=26) received treatment regimens involving polypharmacy with SGAs. These patients as a group achieved statistically significant reductions on their scores on the Brief Psychiatric Rating Scale (34.2 +/- 11.0 compared with 25.3 +/- 11.8; p=0.016) and the Clinical Global Impressions-Improvement Scale (5.5 +/- 0.6 compared with. 5.0 +/- 0.8; p=0.016) at 6 months. There was a significant decrease in the use of prn medications (7.6 +/- 19.6 compared with 1.6 +/- 2.6; p<0.04). However, the number of patients receiving anticholinergic medications increased from 5 to 8 (p<0.04). CONCLUSIONS: Polypharmacy with SGAs is quite frequent among chronic inpatients with severe and persistent mental illness despite a limited empirical database supporting its use. The results of our pilot study do not demonstrate the effectiveness and safety of this practice. However, methodological shortcomings may have contributed to our failure to detect a true, positive effect. Controlled studies are needed to accurately determine the risks and benefits of SGA polypharmacy. PMID- 17414693 TI - Clinical predictors of renal allograft histopathology: a comparative study of single-lesion histology versus a composite, quantitative scoring system. AB - BACKGROUND: Progressive injury that is refractory to conventional immunosuppression remains the major hurdle to indefinite survival of transplanted organs. Several clinical risk factors of chronic renal allograft rejection have been identified; although some (e.g., acute rejection) are direct manifestations of immunological injury, others (e.g., donor age) have been more difficult to conceptually link with graft dysfunction. METHODS: We conducted formal multivariate statistical analyses to reveal associations between established clinical risk factors and allograft histopathology. In a multicenter protocol biopsy-controlled study, 17 clinical risk factors were studied in relation to either the composite Chronic Allograft Damage Index (CADI) score or, to each of eight individual histological indices, using multiple linear regression with forward selection. RESULTS: Nine clinical risk factors were not significantly associated with any histopathological index. Four (donor age, acute rejection, recipient age, and cold ischemia time) were associated both with the total CADI score and, to varying extents, with the individual histopathological indices. In our analysis, clinical risk factors accounted for, at best, only about 60% of the interindividual variation in histopathological score. CONCLUSIONS: Our study reveals a missing link between specific clinical risk factors and early histopathological findings that are known to presage accelerated failure of clinically healthy grafts. Given the complex relationship between clinical risk factors, early histopathological changes, and graft outcome, we conclude that composite, quantitative histological indices are best suited to for evaluation of the histological status of the transplant. PMID- 17414694 TI - CADI, Canti, Cavi. PMID- 17414695 TI - Risk factors, graft fibrosis, and outcome: making the links. PMID- 17414696 TI - Predicting allograft survival: abundant data, but insufficient knowledge? PMID- 17414697 TI - Importance of age in evolving graft histology. PMID- 17414698 TI - The matrix comes to lung transplantation. AB - Lung transplantation is complicated by fibroproliferation, which is likely mediated in part by matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs. This commentary briefly discusses what is known about these mediators in fibrotic pulmonary diseases and how an important new study by Yoshida and colleagues sheds light on the diverse functions of these proteins in alloimmune inflammation. PMID- 17414699 TI - Generation of functional islet-like clusters after monolayer culture and intracapsular aggregation of adult human pancreatic islet tissue. AB - BACKGROUND: Cellular replacement therapy represents a promising strategy for treating type I diabetes; however, such an approach is limited due to the inadequate availability of human donor tissue. Here we investigated the extent to which human islet tissue can be expanded in monolayer culture and brought back to islet function. METHODS: Adult human pancreatic cells were proliferated with a serum-free media in monolayer cultures through multiple passages. Expanded cells were dispersed and encapsulated in alginate-poly-l-lysine microcapsules wherein the cells spontaneously coalesced into islet-like clusters. Encapsulated cell clusters were subsequently transplanted into the peritoneal cavity of streptozotocin-induced diabetic severe combined immunodeficiency mice. RESULTS: The cultured monolayer cells secreted insulin in response to glucose stimulation and maintained endocrine gene expression. Encapsulated islet-like clusters displayed cellular architecture similar to freshly isolated and encapsulated adult human islets maintained in culture, exhibiting an immunoreactive core of insulin, glucagon, and somatostatin, as well as peripheral cytokeratin-19 staining. Encapsulated aggregates significantly reduced hyperglycemia in transplanted mice within 1 week and normoglycemia was achieved after 5 weeks. Human C-peptide was detected in transplanted mice concomitant with the reduction in hyperglycemia. Capsules recovered 8 weeks posttransplantation exhibited insulin immunoreactivity. CONCLUSIONS: Collectively, these data indicate that adult human pancreatic islet cells can be expanded by three serial passages while maintaining their endocrine properties and can yield functional islet-like cell clusters through intracapsular aggregation that reverse hyperglycemia in diabetic mice. This culture and aggregation process could serve as a platform for proliferation and differentiation studies of endocrine lineage cells. PMID- 17414700 TI - Retinal complications in patients with solid organ or bone marrow transplantations. AB - BACKGROUND: The administration of systemic immunosuppressive agents to recipients of solid organ or bone marrow transplants results in an immunocompromised status. As the number of organ recipients and their life span increase with recent progress in organ transplantation, ocular complications tend to become more diverse and serious. METHODS: From 1995 to 2005, 3656 cases of organ transplantations were performed at Asan Medical Center. The medical records of 1198 of these patients who had been examined at the Department of Ophthalmology were reviewed. RESULTS: Retinal complications were diagnosed in 33 of the transplant recipients; five with bone marrow transplantation, 16 with kidney transplantation, seven with liver transplantation (LT), and five with heart transplantation. Diagnoses included 11 cases of CMV (cytomegalovirus) retinitis, three of acute retinal necrosis, one of progressive outer retinal necrosis, five of fungal chorioretinitis, one of toxoplasmic retinochoroiditis, three of central retinal vein occlusion, and nine of central serous chorioretinopathy. While CMV, fungal, or toxoplasmic chorioretinitis developed frequently in association with extraocular infection or organ rejection, herpetic infection manifested only in the eye without any rejection. Most infectious cases responded well to the standard therapeutic regimen. Interestingly, central retinal vein occlusion developed exclusively following LT, possibly in relation to coagulation cascade abnormality. CONCLUSIONS: To our best knowledge, this comprehensive review presents the largest series of ocular complication in organ transplant recipients. Familiarity with the potential ocular complications as well as a high index of suspicion is warranted to the practicing ophthalmologists. PMID- 17414701 TI - Changes in coronary arterial dimensions early after cardiac transplantation. AB - BACKGROUND: Significant changes in coronary artery structure, including intimal thickening and vessel remodeling, occur early after cardiac transplantation. The degree to which these changes compromise coronary lumen dimensions, and the clinical factors that affect these changes, remain controversial. METHODS: Thirty eight adult cardiac transplant recipients underwent coronary angiography and volumetric intravascular ultrasound (IVUS) evaluation of the left anterior descending artery within 8 weeks of transplantation and at 1 year. Clinical parameters including donor and recipient characteristics, rejection episodes, and serology were prospectively recorded. Two-dimensional IVUS measurements and vessel, lumen and plaque volume were calculated at both time points and compared. Multivariate regression analysis was performed to reveal clinical predictors of change in coronary dimensions. RESULTS: During the first year after transplantation, significant decreases in vessel size (negative remodeling) and lumen size were observed with significant increases in plaque burden based on IVUS analyses. Loss of lumen volume correlated significantly with the degree of negative remodeling (R=0.82, P<0.0001), but not with changes in plaque burden (R=0.08, P=0.64). Patients with the greatest increase in plaque volume had significantly less negative remodeling (R=0.53, P=0.0006). Transplant recipient cytomegalovirus (CMV) antibody seropositivity and lack of aggressive prophylaxis against CMV infection/reactivation were significant independent predictors of greater negative remodeling (P<0.01 and P=0.03, respectively) and greater lumen loss (P=0.02 and P=0.03, respectively). CONCLUSION: Negative remodeling is primarily responsible for coronary artery lumen loss during the first year after cardiac transplantation. CMV seropositivity and lack of aggressive CMV prophylaxis correlate with increased negative remodeling, resulting in greater lumen loss. PMID- 17414702 TI - Association of elevated pretransplant sCD30 levels with graft loss in 206 patients treated with modern immunosuppressive therapies after renal transplantation. AB - BACKGROUND: Recent reports suggest that high pretransplant serum levels of soluble CD30 (sCD30) are a risk factor for rejections after kidney transplantation. The aim of our study was to elucidate the predictive value of pretransplant sCD30 levels for kidney transplantation outcome in a single-center patient cohort that has been treated with modern immunosuppressive therapies after transplantation. METHODS: We retrospectively analyzed sCD30 in multiple pretransplant sera from 206 patients, of whom 174 were transplanted with a cadaveric kidney and 32 patients received an allograft from a living donor. Renal function after transplantation was estimated by measuring serum creatinine and by rejection diagnosis. RESULTS: We could demonstrate a statistically significant association between increased pretransplant sCD30 values and graft failures (P=0.005). Receiver operating curve analysis revealed a cutoff value of 124 U/mL pretransplant sCD30. A multivariate analysis confirmed pretransplant sCD30 values >124 U/mL (P=0.011) and rejection episodes (P<0.0001) as independent risk factors for graft loss. CONCLUSION: Our study revealed a strong correlation between pretransplant sCD30 levels and the incidence of graft failure, but we could not confirm that the development of rejection episodes is correlated with pretransplant sCD30 values. PMID- 17414703 TI - Long-term cell monitoring of kidney recipients after an antilymphocyte globulin induction with and without steroids. AB - BACKGROUND: Because of several side effects, the corticosteroid usage has been minimized in kidney transplantation. The increased acute rejection episodes associated with their withdrawal may counterbalance with induction treatment using polyclonal antilymphocyte globulin (ALG). The effects of ALG on blood cell phenotype have already been the subject of several reports. However, to date, no data are available concerning the comparison of blood phenotype when ALG is given with or without steroids and no gene profiling study has been performed. METHODS: We report here on a longitudinal blood cell analysis of a selected cohort of kidney recipients enrolled in a randomized study of steroid avoidance or withdrawal (during 6 months) during ALG induction. RESULTS: In the two groups, ALG quickly and massively depleted all the T cells and natural killer cells, but not B cells. Interestingly, the lymphopenia-driven homeostatic proliferation of CD4 and CD8T cells strongly differed with persistent low CD4 (including CD25CD4) T-cell counts. Effector memory CD8T cells reappeared rapidly. ALG induced apoptosis-associated molecules and increased myeloid cell genes. However, few genes were found differentially expressed with a low fold ratio between the two groups during and at distance of corticotherapy. CONCLUSION: Thus initial steroid avoidance or withdrawal associated with ALG induction has a weak influence on phenotype and transcriptional pattern of blood leukocytes. In contrast, ALG therapy induces an early and strong depletion of all T-cell subsets with contrasted long-lasting homeostatic regulation. PMID- 17414704 TI - Stable kidney function in the second decade after kidney transplantation while on cyclosporine-based immunosuppression. AB - BACKGROUND: Calcineurin inhibitors (CNIs) have been the mainstay of immunosuppressive protocols in kidney transplantation over the past 20 years. However, in some recipients, the adverse effects of CNIs contribute to chronic allograft nephropathy and death with function--the two leading causes of late graft loss. Other recipients maintain stable graft function. METHODS: We studied the impact of continuing CNI-based immunosuppression in the second decade after kidney transplantation. From 1984 through 1996, a total of 1,263 patients underwent a primary kidney transplant at the University of Minnesota and received cyclosporine-based immunosuppression. Antibody induction was used only in deceased donor recipients. RESULTS: The actuarial 20-year patient survival rate was 38%; graft survival, 30%; and death-censored graft survival, 60%. The annual mean serum creatinine level for recipients whose grafts survived > or =1 year remained stable, although recipients with a history of > or =1 acute rejection episode had a higher serum creatinine level vs. recipients who were rejection free. The annual mean calculated creatinine clearance was also stable over time. In addition, for recipients who were acute rejection-free, chronic allograft nephropathy/chronic rejection was only responsible for 9% of graft losses. CONCLUSIONS: Our study suggests that some kidney transplant recipients tolerate long-term CNI-based immunosuppression with stable creatinine levels. Identifying certain recipients' predisposition to CNI toxicity and individualizing immunosuppressive therapy may be important in order to improve long-term kidney function, while simultaneously preserving low short-term acute rejection rates. PMID- 17414705 TI - Determinants of the complement-fixing ability of recipient presensitization against HLA antigens. AB - BACKGROUND: The presence of preformed alloantibodies with the ability to activate complement may pose a particular risk for kidney allograft rejection. The aim of this study was to evaluate variables that determine the complement-fixing capability of human leukocyte antigen (HLA) sensitization. METHODS: Sixty-five sensitized patients with > or =10% pretransplant panel-reactive antibody (PRA) levels uncovered by immunoglobulin G [IgG]FlowPRA HLA class I and/or class II screening were included. Applying modified FlowPRA screening, sera were evaluated for patterns of alloreactive IgG subclasses and IgM, and, in parallel, for their complement-activating ability assessed by flow cytometric detection of human complement split product deposition ([C4d]FlowPRA). RESULTS: Approximately two thirds (68%) of tested sera were found to contain complement-fixing alloreactivity (> or =10%[C4d]FlowPRA). IgG1 type panel reactivity was predominant (detectable HLA class I and II reactivity in 93% and 91% of IgG positive sera), followed by IgG3 (49%/44%), IgG2 (44%/27%), and IgG4 (19%/11%). Applying partial correlation we found an independent correlation of both %[IgG1]FlowPRA and %[IgG3]FlowPRA with %[C4d]FlowPRA reactivities (P< or =0.01). In addition, for IgG1 a contribution of the amount of bound alloantibody to complement-fixation was observed. Complement-fixation was also favored by the simultaneous presence of alloreactive IgG1, IgG3, and IgM. Previous grafting, but not pregnancy and transfusion, was independently associated with complement fixing sensitization (P<0.05), presumably due to increased IgG1 type reactivity. CONCLUSIONS: Anti-HLA antibody-triggered complement activation is dependent on both the pattern of Ig reactivities and the amount of bound antibody. Previous transplantation represents a major risk factor for the development of complement fixing sensitization. PMID- 17414706 TI - Allotransplantation of cultured parathyroid progenitor cells without immunosuppression: clinical results. AB - BACKGROUND: Hypoparathyroidism is a well-known consequence of extensive thyroid and parathyroid surgery. Allotransplantation of cultured parathyroid cells can be considered as an alternative to vitamin D3 and calcium supplementation in treatment of hypoparathyroidism. We present the long-term allotransplant activity in 85 patients who had undergone cellular allotransplantation for surgical hypoparathyroidism. Also, a modified technique to prepare parathyroid explants is described for obtaining a new nonimmunogenic cell population. METHODS: From March 1990 to December 2004, 85 patients underwent 116 allotransplantations of cultured parathyroid cells. Mean recipient age was 46.2+/-11.1 years. Donors were selected from patients undergoing parathyroidectomy for secondary and tertiary hyperparathyroidism. RESULTS: After 6 weeks of cultivation and freezing, the parathyroid cells decreased their normal human leukocyte antigen (HLA) class I ABC expression and were free of HLA class II positive cells. The viability of cultured cells was 95.15+/-2.94%. Eighty-five patients underwent primary allotransplantation. Of these, 25 patients subsequently underwent a repeat procedure. In six cases, the parathyroid cells were obtained from the same donor and in 19 cases from a different donor. For all patients, the mean cellular allograft survival was 6.35+/-13.08 months. In 64 patients (55.1%), the allografts retained their endocrine function for more than 2 months. CONCLUSIONS: The present study has shown that in some patients parathyroid cell allotransplantation may be considered a method of treatment for permanent hypoparathyroidism after thyroid surgery. Graft function and/or survival did not depend on the baseline viability or secretory activity of cultured cells used for transplantation. PMID- 17414707 TI - The effect of FK778 on the progression of chronic allograft nephropathy in a rat model. AB - BACKGROUND: Chronic allograft nephropathy (CAN) still belongs to the leading causes of graft loss over the long term. The leflunomide derivative FK778 is a novel immunosuppressant with improved pharmacokinetic properties that effectively prolonged graft survival in several transplantation models. In the present study, we investigated the effects of FK778 at different phases after transplantation on the progression of CAN. METHODS: Fisher 344 kidneys were orthotopically transplanted into Lewis recipients. Recipients were treated with FK778 (5 mg/kg/day) over different time periods (early: days 0-10 only, continuous: day 0 to week 24, or late: weeks 16-24 only posttransplantation). Proteinuria was measured every 4 weeks, whereas grafts were harvested at 24 weeks posttransplantation for morphological and immunohistochemical analysis as well as transforming growth factor-beta and platelet derived growth factor-B chain expression. RESULTS: Continuous treatment with FK778 ameliorated the progression of CAN, whereas late treatment reduced proteinuria and resulted in a similar grade of CAN as compared to animals with continuous treatment. In contrast, FK778 given only during the early phase after transplantation had no effect on the progression of CAN as compared to controls. CONCLUSION: In summary, FK778 is a potent immunosuppressive drug that can delay the progression of CAN, even when given at later stages after transplantation. PMID- 17414708 TI - Riboflavin-mediated reduction of oxidant injury, rejection, and vasculopathy after cardiac allotransplantation. AB - BACKGROUND: Riboflavin is a well-known nutritional supplement that has been shown to exhibit antioxidant properties and protect tissue from oxidative damage. We hypothesized that riboflavin given during cardiac ischemia-reperfusion (I/R) might reduce subsequent acute rejection, after allotransplantation, and coronary allograft vasculopathy (CAV). METHODS: A murine heterotopic cardiac transplantation model was used to test whether riboflavin improves I/R injury and acute/chronic rejection. RESULTS: Riboflavin significantly reduced oxidant production and inflammatory mediator production induced by I/R injury, as evidenced by decreased levels of malondialdehyde, myeloperoxidase activity, and tumor necrosis factor alpha. Administration of riboflavin also improved graft survival and suppressed T-cell infiltration and donor-reactive alloantibody formation during the early period after allotransplantation. A murine long-term cardiac allograft model using immunosuppression (preoperative anti-murine CD4 and anti-CD8) was employed to investigate the effect of riboflavin against CAV at 60 days. Riboflavin-treated grafts exhibited a significant decrease in the severity of coronary artery luminal occlusion as compared with saline-treated grafts (17.4+/-1.8% vs. 43.5+/-5.6%, P=0.0012). However, there was no significant effect of riboflavin to reduce donor-reactive alloantibodies in this chronic model. CONCLUSIONS: These data indicate that riboflavin improves early I/R injury and reduces the development of CAV, most likely due to alloantigen-independent effects such as reduced early graft oxidant stress. Riboflavin administered in the setting of cardiac allograft transplantation appears to be a powerful means to reduce early graft lipid peroxidation, leukocytic infiltration, and cytokine production as well as to suppress the late development of cardiac allograft vasculopathy. PMID- 17414709 TI - Application of the two-layer method on pancreas digestion results in improved islet yield and maintained viability of isolated islets. AB - BACKGROUND: Oxygenation of the pancreas during preservation by the two-layer method (TLM) has shown beneficial effects in islet transplantation. Here, we apply this concept (oxygenation) to the isolation process. METHODS: Rat pancreases were digested using four different methods. Pancreases were digested with preoxygenated perfluorocarbon (PFC) in group 2 and without it in group 1. Additionally, adenosine was included in the collagenase solution in subgroups B but not in subgroups A. Islet yields and viability were compared between groups. RESULTS: Tissue oxygen tension in group 1 was essentially zero during digestion, but rapidly reached around 300 mm Hg and was maintained in group 2. The tissue adenosine triphosphate (ATP) level in rat pancreas just after laparotomy (control) was 4.2+/-0.7 micromol/g dry weight; after digestion, it was 0.12+/ 0.03 micromol/g, 0.70+/-0.10 micromol/g, 0.30+/-0.18 micromol/g, and 2.90+/-0.80 micromol/g in groups 1A, 1B, 2A, and 2B, respectively. No significant differences were observed between group 2B and control (P=0.19). Islet yields (IEQ/pancreas) were 1600+/-400, 1400+/-400, 1300+/-400, and 2400+/-100 in groups 1A, 1B, 2A, and 2B, respectively. The islet yield of group 2B was significantly higher than other groups (P<0.05). The cure rate after transplanting 200 islets into athymic nude mice did not differ (80% in all groups). The stimulation indices in the four groups were also the same. CONCLUSIONS: Tissue ATP levels after digestion were well maintained using TLM with adenosine digestion method. Consequently, greater numbers of islets could be retrieved. The new method was at least equivalent to islet function isolated by conventional method. Clinical study is therefore warranted. PMID- 17414710 TI - Saliva secretion stimulated by grafted nerve in submandibular gland allograft in dogs. AB - BACKGROUND: Recent studies have described submandibular gland allografts in animal models; however, the amount of saliva secretion or nervous regeneration in those animals have not been reported. Herein, we investigated saliva secretion from submandibular gland allografts in beagle dogs. METHODS: Using a vascularized submandibular gland transplantation method, we extracted portions of the submandibular gland including the duct from beagle dogs and placed them into the submental region of age- and weight-matched dogs. Differences in the amount of saliva secretion and histologic appearance were compared based on the existence of chorda tympani branches in the allograft. RESULTS: At 10 weeks after transplantation, the amount of resting saliva in the group grafted with the nerve was clearly increased, whereas the quantity of that in the group that underwent transplantation without the nerve was quite low. In the former group, responses were demonstrated after taste stimulation and electronic nerve stimulation. CONCLUSIONS: Our results showed a clear relationship between the presence of a nerve in grafted submandibular glands and saliva secretion. PMID- 17414711 TI - Transplant tolerance associated with a Th1 response and not broken by IL-4, IL-5, and TGF-beta blockade or Th1 cytokine administration. AB - BACKGROUND: Specific transplant tolerance is mediated by CD4 T cells that die unless supported by T-cell derived cytokines and donor antigen. This study examined the role of Th1 and Th2 cytokines in the maintenance of tolerance. METHODS: Tolerance to fully allogeneic PVG cardiac allografts in DA rats was induced by short-term anti-CD3 monoclonal antibody therapy. Responses of tolerant cells to donor and third party antigen were assessed in vivo by examination of the infiltrate in the heart and application of skin grafts, and in vitro in mixed lymphocyte culture. Cell subsets were stained, induction of cytokine mRNA assayed by reverse-transcriptase polymerase chain reaction and the role of cytokines determined by treating with blocking monoclonal antibody to cytokines or cytokine administration. RESULTS: Tolerated grafts had a T cell and macrophage infiltrate with increased mRNA for Th1 cytokines, interleukin (IL)-2, and interferon (IFN) gamma but not Th2 cytokines. Peripheral lymphocytes proliferated in mixed lymphocyte culture and expressed Th1 cytokine mRNA. Tolerant hosts accepted PVG and rejected Lewis skin allografts and the lymph nodes draining both these grafts had similar induction of Th1 and Th2 cytokine mRNA. Treatment of tolerant rats with Th1 cytokines IL-2, IFN-gamma, and IL-12p70 or monoclonal antibody that blocked IL-4, IL-5, and transforming growth factor-beta did not prevent acceptance of PVG skin grafts. CONCLUSIONS: These studies in a model of tolerance regulated by CD4CD25 T cells demonstrated there was no defect in Th1 responses. Tolerance was due to regulation that was not solely dependent on IL-4, IL-5, or transforming growth factor-beta and was not inactivated or overwhelmed by administration of Th1 cytokines, IL-2, IFN-gamma or IL-12p70. PMID- 17414712 TI - Involvement of the programmed death-1/programmed death-1 ligand pathway in CD4+CD25+ regulatory T-cell activity to suppress alloimmune responses. AB - BACKGROUND: Immune regulatory CD4+CD25+ T (regulatory T; Treg) cells play a vital role in the induction and maintenance of self-tolerance. They are essential for the homeostasis of T cells, the prevention of autoimmunity, and the induction of tolerance to allogeneic donor grafts. However, the underlying mechanism of their functions remains mostly elusive. Therefore, we investigated here a crucial role of Treg cells in their response to alloantigen via the programmed death (PD)-1/PD 1 ligand (PD-L1) pathway. METHODS: In vitro mixed lymphocyte reaction (MLR) assay, graft-versus-host disease (GvHD) and a skin transplantation model were used to evaluate the mechanisms of PD-1/PD-L1 pathway. RESULTS: Blockade of the PD-1/PD-L1 pathway using anti-PD-L1 monoclonal antibodies (mAb) is found to inhibit Treg cell's ability to suppress and restore CD4+CD25-T-cell proliferation in vitro. GvHD was lethal after adoptive transfer of allogeneic C57BL/6 (H-2K) spleen cells to NOD/SCID (H-2K) mice unless CD25+ T cells were also included. Strikingly, the suppression of GvHD by CD25+ cells was abrogated by anti-PD-L1 mAb administration. The abrogation of Treg-cell-mediated suppression could also be demonstrated in a Balb/c (H-2K) to B6/Rag-2KO (H-2K) skin-allograft model. CONCLUSIONS: The blockade of the PD-1/PD-L1 pathway abrogates Treg-mediated immunoregulation, thus suggesting that the PD-1/PD-L1 pathway is required for Treg suppression of the alloreactive responses of CD4+CD25-T cells. This finding has important implications for clarifying the mechanisms of allograft rejection and GvHD. PMID- 17414713 TI - Although pig allogeneic mesenchymal stem cells are not immunogenic in vitro, intracardiac injection elicits an immune response in vivo. AB - BACKGROUND: In vitro, mesenchymal stem cells (MSCs) have demonstrated a low immunogenic profile. In this study, we tested the immune response to allogeneic MSCs in immunocompetent swines both in vitro and in vivo. METHODS: Major histocompatibility complex-controlled swine leukocyte antigen (SLA) and SLA were used as donor and recipient, respectively. In vitro, proliferative responses were tested by mixed lymphocyte reaction (MLR) or cocultures and cytokine profiling by enzyme-linked immunosorbent assay. In vivo, allogeneic MSCs were injected in cardiac infarcted area (n=3) and compared with subcutaneous injections of either MSCs (n=2) or peripheral blood mononuclear cells (PBMCs; n=2). Two additional animals received a skin graft as controls. No immunosuppression was used. Specific antidonor humoral responses were tested by flow cytometry and complement dependent cytotoxicity assay. RESULTS: In vitro, either unstimulated MSCs or interferon (IFN)-gamma stimulated MSC failed to elicit a proliferative response (stimulation index: 1.23 vs. 1.12 vs. 36.9 for controls, P<.001). Concomitantly to the absence of proliferation to MSCs, low production of IFN-gamma and interleukin-2 was evidenced in supernatants while the production of Th2 cytokines was comparable to controls. In vivo, all animals receiving skin grafts, subcutaneous PBMCs and intracardiac MSCs injections developed donor-specific cellular and humoral responses (immunoglobulins M and G) with antibody-complement mediated cytotoxicity. Subcutaneous MSCs injection needed a rechallenge to similarly develop a cytotoxic humoral response. CONCLUSIONS: Intracardiac allogeneic porcine mesenchymal stem cells elicit an immune response despite their low immunogenic profile in vitro. This result suggests that in vivo characteristics of allogeneic MSCs might differ and emphasizes the importance of pursuing research both in vitro and in vivo. PMID- 17414714 TI - Diagnosis of acute renal allograft rejection by analyzing whole blood mRNA expression of lymphocyte marker molecules. AB - BACKGROUND: Currently, the diagnosis of acute rejection after kidney transplantation is based on a kidney biopsy taken after clinical rejection suspicion. A robust, noninvasive diagnostic method would allow easier and more frequent monitoring of the patient and the graft. Potentially, a straightforward method would be the analysis of lymphocyte marker molecule expression from whole blood samples. METHODS: Whole blood samples were collected prospectively in a single kidney transplantation center from 50 adult kidney recipients transplanted between 2001 and 2005. The mRNA expression of granzyme B, perforin, FasL, granulysin, CD154, ICOS, CTLA4 and PD-1 were analyzed with real-time quantitative polymerase chain reaction. RESULTS: The expression of ICOS and CD154 were significantly lower in rejection patients than in control patients (P<0.001). Both genes gave statistically significant area under receiver operating characteristic curve (AUC; 0.87, 0.88) with 84% sensitivity and 100% specificity for CD154 and 76% and 86% for ICOS, respectively. In paired rejection and postrejection therapy samples, the expression of both genes significantly increased during rejection therapy (P<0.001). When rejection patients were compared to patients biopsied because of other reasons of graft dysfunction, both CD154 and ICOS were lower in rejection patients but only CD154 was statistically significant (P=0.028, AUC=0.740, sensitivity 52%, specificity 90%). The other studied genes gave no consistent statistically significant results. CONCLUSIONS: The whole blood gene expression quantities of costimulatory molecules CD154 and ICOS reasonably robustly differentiated rejection patients from control patients. The clinical use of the analysis is limited by poor capability to differentiate patients with rejection from patients with other causes of graft dysfunction. PMID- 17414715 TI - Metalloproteinase inhibition has differential effects on alloimmunity, autoimmunity, and histopathology in the transplanted lung. AB - BACKGROUND: Upregulation of matrix metalloproteinases (MMPs) has been associated with chronic lung allograft rejection known as bronchiolitis obliterans syndrome. It has been suggested that MMP inhibition could prevent the rejection response. However, the effect of MMP inhibition on lung allograft rejection has not been reported. METHODS: Utilizing a rat model of lung transplantation, tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) were overexpressed by gene therapy in F344 rat lung allografts prior to transplantation into WKY recipient rats. Separately, WKY rats that received F344 lung allografts were treated systemically with COL-3, a global MMP inhibitor. RESULTS: TIMP-1 and TIMP-2 had differential effects on delayed type hypersensitivity (DTH) responses to donor antigens and type V collagen, an autoantigen involved in the rejection response. Neither TIMP-1 or TIMP-2 affected the onset of rejection pathology. COL-3 suppressed DTH responses to donor antigens and type V collagen, abrogated local production of tumor necrosis factor-alpha, and interleukin-1beta. Although it did not prevent rejection pathology, COL-3 (30 mg/kg) induced intragraft B cell hyperplasia suggestive of posttransplant proliferative disorder (PTLD). CONCLUSIONS: These data identify a complex role for MMPs and TIMPs in the immunopathogenesis of lung allograft rejection, and indicate their effects are not limited to matrix remodeling. PMID- 17414716 TI - Epithelial-to-mesenchymal transition of human proximal tubular epithelial cells: effects of rapamycin, mycophenolate, cyclosporin, azathioprine, and methylprednisolone. AB - Epithelial-to-mesenchymal transition (EMT) of renal proximal tubular epithelial cells (PTEC) into myofibroblasts is an important step in the pathogenesis of chronic allograft nephropathy. The effects of commonly used immunosuppressives in renal transplantation on EMT are not known. PTEC were cultured in transforming growth factor-beta to induce EMT. The effects of the immunosuppressives on cell morphology and alpha-smooth muscle actin were studied by phase contrast microscopy, immunocytochemistry, and western blotting. The effects on versican were studied by [S] labeling and polyacrylamide gel electrophoresis. Rapamycin and mycophenolate mofetil (MMF) prevented EMT and moreover returned myofibroblasts to PTEC morphology. These immunosuppressives also reduced versican production by both PTEC and myofibroblasts. Cyclosporine A, azathioprine, and methylprednisolone were less effective than rapamycin and MMF. Moreover, these immunosuppressives did not decrease versican. Rapamycin and MMF have a greater inhibitory effect on EMT in vitro than older immunosuppressives and may result in less fibrosis and a better long-term allograft survival. PMID- 17414717 TI - Donor T-cell development in host thymus after heterotopic limb transplantation in mice. AB - We developed a mouse model of heterotopic limb transplantation in which we took advantage of Thy1.1 and Thy1.2 congenic strains to track and characterize donor T cells, to determine the role of recipient's thymus in mixed T-cell chimerism induction as well as transplant immunocompetence. The vascularized Thy1.1 limb graft composed of femur, muscle, and skin (VBT) survived long-term in more than 87.5% of Thy1.2 recipients. Percentages of donor-type Thy1.1 T cells increased from day 30 to 90 in thymus and spleen of recipients. Most peripheral donor T cells displayed a naive phenotype and a few others were regulatory T cells. Thymectomy prevented peripheral T-cell chimerism. Congenic VBT in immunodeficient RAG mice restored their ability to reject skin allografts. These observations suggest that donor T cells differentiated in host thymus might contribute to maintenance of mixed chimerism after transplantation of tissue composite grafts that include vascularized bone. PMID- 17414718 TI - Intrahepatic detection of FOXP3 gene expression after liver transplantation using minimally invasive aspiration biopsy. AB - Intragraft accumulation of Forkhead box P3 (FOXP3)-positive regulatory T cells (Treg) is associated with local suppression of alloresponses in transplantation models. In the current study, the utility of the minimally invasive fine needle aspiration biopsy for the intragraft detection of FOXP3 and interferon (IFN) gamma mRNA expression was investigated in clinical liver transplantation (LTx). Intragraft FOXP3 increased within the first year after LTx, but not in blood. Elevated FOXP3, but not IFN-gamma expression, in the liver was observed after hepatitis C virus (HCV) reinfection and after a previous episode of acute rejection. These data show the feasibility of aspiration biopsy for intragraft monitoring of gene expression. Intrahepatic FOXP3 levels are associated with HCV reinfection, a history of acute rejection, and increased within the first year after LTx. Differences in gene expression between the graft and blood underline the importance of local immune monitoring. PMID- 17414719 TI - Dapsone-induced artifactual a1c reduction in islet transplant recipients. PMID- 17414720 TI - Venous sinus thrombosis in a renal transplant patient. PMID- 17414721 TI - Transplantation of dual adult kidneys into a recipient with minimal abdominal vascular access. PMID- 17414722 TI - Transplantation of a horseshoe kidney. PMID- 17414723 TI - Applicability of a nomogram to predict DGF. PMID- 17414724 TI - Proinsulin c-peptide exerts beneficial effects in endotoxic shock in mice. AB - OBJECTIVE: Insulin connecting peptide (c-peptide) aids the folding of proinsulin and has been considered to have little biological activity. Recently, c-peptide has been shown to improve diabetic neuropathy and nephropathy as well as vascular inflammation. In vitro studies have reported that c-peptide may activate peroxisome proliferator-activated receptor-gamma, a nuclear transcription factor that plays a regulatory role in inflammation. This study was designed to investigate the biological effects of c-peptide during endotoxemia. DESIGN: Prospective, randomized laboratory investigation that used an established murine model of endotoxic shock. SETTING: University hospital laboratory. SUBJECTS: Mice were subjected to endotoxic shock by intraperitoneal administration of Escherichia coli lipopolysaccharide. INTERVENTIONS: Mice received vehicle or c peptide (70-140 nmol/kg) intraperitoneally at 3 hrs and 6 hrs after lipopolysaccharide. Mortality was monitored for 96 hrs. In a separate experiment, mice were killed at 4, 7, and 18 hrs after lipopolysaccharide administration. Lungs and plasma were collected for biochemical assays. MEASUREMENTS AND MAIN RESULTS: In vehicle-treated mice, endotoxic shock resulted in lung injury and was associated with a 41% survival rate and elevation in plasma tumor necrosis factor alpha, macrophage inflammatory protein-1alpha, monocyte chemoattractant protein 1, and keratinocyte-derived chemokine levels. Lung nuclear levels of phosphorylated extracellular signal-regulated kinases 1 and 2 were significantly increased in vehicle-treated mice. On the other hand, lung nuclear expression and DNA binding of proliferator-activated receptor-gamma were decreased in comparison to control animals. Treatment with c-peptide (140 nmol/kg) improved survival rate (68%) and reduced plasma levels of tumor necrosis factor-alpha, macrophage inflammatory protein-1alpha, and monocyte chemoattractant protein-1, but it did not exert hypoglycemic effects. Treatment with c-peptide also up-regulated lung nuclear expression and DNA binding of proliferator-activated receptor-gamma and reduced phosphorylation of extracellular signal-regulated kinases 1 and 2 in comparison to vehicle-treated mice. CONCLUSIONS: Our data show that c-peptide has beneficial effects in endotoxic shock, and this therapeutic effect is associated with activation of proliferator-activated receptor-gamma. PMID- 17414725 TI - Sepsis incidence and outcome: contrasting the intensive care unit with the hospital ward. AB - OBJECTIVE: To describe the outcome of patients with sepsis according to location on a ward or in an intensive care unit. DESIGN: Prospective multicentered observational study. SETTING: Three academic hospitals in Madrid, Spain. PATIENTS: Consecutive patients with sepsis admitted to participating hospitals from March 1 to June 30, 2003. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During the study period, 15,852 patients >18 yrs of age were admitted. Sepsis was identified in 702 patients, giving an estimated cumulative incidence rate of 367 cases per 100,000 adult area residents per year and a cumulative incidence rate among patients admitted to the hospital of 4.4%. Most septic patients had a community-acquired infection (71%). Severe sepsis developed in 199 patients (incidence rate, 104 cases per 100,000 adult area residents per year), and 59 patients developed septic shock (incidence rate, 31 cases per 100,000 adult area residents per year). Most of the patients met the criteria for severe sepsis or septic shock on the same day that they would have qualified for the septic status one step down the scale. In the other patients, the median time between sepsis and severe sepsis was 2 days (interquartile range, 2-5) and between severe sepsis and septic shock was 3 days (interquartile range, 1-4). Only 32% of severe sepsis patients received intensive care. The hospital mortality for all septic patients was 12.8%; for severe sepsis, 20.7%; and for septic shock, 45.7%. CONCLUSIONS: This study shows the high incidence of sepsis in a general population of patients admitted to hospital. A significant proportion of patients with severe sepsis are not transferred to the intensive care unit. PMID- 17414726 TI - Video-based training increases sterile-technique compliance during central venous catheter insertion. AB - OBJECTIVE: To evaluate the effect of an online training course containing video clips of central venous catheter insertions on compliance with sterile practice. DESIGN: Prospective randomized controlled study. SETTING: Admitting area of a university-based high-volume trauma center. SUBJECTS: Surgical and emergency medicine residents rotating through the trauma services. INTERVENTIONS: An online training course on recommended sterile practices during central venous catheter insertion was developed. The course contained short video clips from actual patient care demonstrating common noncompliant behaviors and breaks regarding recommended sterile practices. A 4-month study with a counterbalanced design compared residents trained by the video-based online training course (video group) with those trained with a paper version of the course (paper group). Residents who inserted central venous catheters but received neither the paper nor video training were used as a control group. Consecutive central venous catheter insertions from 12 noon to 12 midnight except Sundays were video recorded. MEASUREMENTS AND MAIN RESULTS: Sterile-practice compliance was judged through video review by two surgeons blinded to the training status of the residents. Fifty residents inserted 73 elective central venous catheters (19, 31, and 23 by the video, paper, and control group operators, respectively) into 68 patients. Overall compliance with proper operator preparation, skin preparation, and draping was 49% (36 of 73 procedures). The training had no effect on selection of site and skin preparation agent. The video group was significantly more likely than the other two groups to fully comply with sterile practices (74% vs. 33%; odds ratio, 6.1; 95% confidence interval, 2.0-22.0). Even after we controlled for the number of years in residency training, specialty, number of central venous catheters inserted, and central venous catheter site chosen, the video group was more likely to comply with recommended sterile practices (p = .003). CONCLUSIONS: An online training course, with short video clips of actual patient care demonstrating noncompliant behaviors, improved sterile-practice compliance for central venous catheter insertion. Paper handouts with equivalent content did not improve compliance. PMID- 17414727 TI - Effect of lower limb compression devices on thermodilution cardiac output measurement. AB - OBJECTIVE: The aim of this study was to determine whether lower limb (calf) sequential compression devices (SCDs) have a significant effect on thermodilution cardiac output measurements using a pulmonary artery catheter. DESIGN: Prospective clinical investigation. SETTING: Surgical and neurosurgical intensive care units in a university hospital. PATIENTS: A total of 43 patients with pulmonary artery catheters and bilateral lower limb SCDs. MEASUREMENTS AND MAIN RESULTS: Cardiac output was measured (average of three) when the SCDs were off (T1), during the first 2-4 secs of the inflation cycle (T2), during seconds 4-8 of the inflation cycle (T3), and when the SCDs were off again (T4). Cardiac output measurements were consistently lower when measured during the SCD inflation cycle. The decrease in cardiac output ranged from 7.58% to 49.5%, with a mean reduction of 24.51% in the first 2-4 seconds and 20.61% during seconds 4-8 (p < .001). Two patients displayed an increase in cardiac output during the inflation cycle; one patient had an increase of 2.78% and the other an increase of 13.5%. In 11 patients, measurements were also made using a pulse contour analysis cardiac output device, but no changes in pulse contour-analysis cardiac output were observed during the same time period. CONCLUSIONS: Thermodilution cardiac output measurements via a pulmonary artery catheter should not be done during the inflation cycle of lower limb SCDs because they produce a falsely low cardiac output. PMID- 17414728 TI - Neutrophil-derived S100A12 in acute lung injury and respiratory distress syndrome. AB - OBJECTIVE: Both persistent accumulation and activation of neutrophils may contribute to the most severe form of acute lung injury, acute respiratory distress syndrome. We analyzed the expression of neutrophil-derived S100A12 and the proinflammatory receptor for advanced glycation end products (RAGE) in patients with acute respiratory distress syndrome. Additional in vivo and in vitro experiments were performed to further analyze the contribution of S100A12 to pulmonary inflammation. SUBJECTS: We included 14 patients with acute respiratory distress syndrome and eight controls. In addition, 16 healthy subjects were included in an experimental lipopolysaccharide challenge model. INTERVENTIONS: Concentrations of S100A12 and soluble RAGE were analyzed in bronchoalveolar lavage fluid. The expression of S100A12 and RAGE in lung biopsies from patients was analyzed by immunohistochemistry. S100A12 was also analyzed in bronchoalveolar lavage fluid from eight healthy subjects after challenge with lipopolysaccharide and compared with eight controls who received placebo inhalation. Effects of S100A12 on endothelial cells were analyzed in vitro. MAIN RESULTS: Patients with acute respiratory distress syndrome had significantly enhanced pulmonary S100A12 expression and higher S100A12 protein concentrations in bronchoalveolar lavage fluid than controls. Levels of soluble RAGE were not significantly elevated in acute respiratory distress syndrome. S100A12 concentrations decreased with time from disease onset. In healthy volunteers, S100A12 was elevated in bronchoalveolar lavage fluid after lipopolysaccharide inhalation. In vitro experiments confirmed strong proinflammatory effects of human S100A12. CONCLUSIONS: S100A12 and its receptor RAGE are found at high concentrations in pulmonary tissue and bronchoalveolar lavage fluid in acute lung injury. S100A12 expression may reflect neutrophil activation during lung inflammation and contribute to pulmonary inflammation and endothelial activation via binding to RAGE. PMID- 17414729 TI - Levosimendan restores both systolic and diastolic cardiac performance in lipopolysaccharide-treated rabbits: comparison with dobutamine and milrinone. AB - OBJECTIVE: Current treatment strategies for severe septic conditions (i.e., intravenous fluids, vasopressors, and cardiac inotropes) reestablish fluid balance and improve cardiac systole but do not address diastolic dysfunction. Our study aimed to fully characterize both systolic and diastolic abnormalities of sepsis-associated heart failure and to identify treatment that would support full cycle cardiac improvement. DESIGN: Endotoxin-injected rabbits, an animal model of abnormal cardiac function in human sepsis, were used to delineate cardiac abnormalities and to examine effects of drug treatments on heart systolic and diastolic function (n = 30); saline-injected animals served as comparators (n = 17). As treatment, three inotropes commonly used for treatment of cardiac failure were infused for 45 mins in separate animal groups-milrinone, dobutamine, and levosimendan. MEASUREMENTS: Variables of left ventricular systolic and diastolic function were assessed with a pressure conductance catheter. Measurements were made before and after endotoxin/saline injection and before and after inotrope treatment. RESULTS: Pressure-volume analyses of the left ventricle showed marked impairment in systolic function and in all indices of diastolic function (isovolumic relaxation time constant, left ventricular end-diastolic pressure, and end-diastolic pressure-volume relationship) in endotoxin-treated rabbits. The inotropes, milrinone, dobutamine, and levosimendan, could each partially or completely restore systolic function in the lipopolysaccharide-treated rabbits. However, only levosimendan therapy led to additional beneficial effects on left ventricular relaxation and diastolic function. CONCLUSIONS: Cardiac failure in severe sepsis results from impairments in both systolic and diastolic functions. Treatment with the calcium sensitizer levosimendan improved both systolic and diastolic cardiac functions in septic animals, but cyclic adenosine monophosphate dependent inotropes milrinone and dobutamine only improved systolic function. PMID- 17414730 TI - Phase II, randomized, controlled trial of high-dose N-acetylcysteine in high-risk cardiac surgery patients. AB - OBJECTIVE: To assess the effect of high-dose N-acetylcysteine on renal function in cardiac surgery patients at higher risk of postoperative renal failure. DESIGN: Multiblind, placebo-controlled, randomized, phase II clinical trial. SETTING: Operating rooms and intensive care units of two tertiary referral hospitals. PATIENTS: A total of 60 cardiac surgery patients at higher risk of postoperative renal failure. INTERVENTIONS: Patients were allocated to either 24 hrs of high-dose N-acetylcysteine infusion (300 mg/kg body weight in 5% glucose, 1.7 L) or placebo (5% glucose, 1.7 L). MEASUREMENTS AND MAIN RESULTS: The primary outcome measure was the absolute change in serum creatinine from baseline to peak value within the first five postoperative days. Secondary outcomes included the relative change in serum creatinine, peak serum creatinine level, serum cystatin C, and in urinary output. Further outcomes were needed for renal replacement therapy, length of ventilation, and length of stay in the intensive care unit and hospital. Randomization was successful and patients were well balanced for preoperative and intraoperative characteristics. There was no significant attenuation in the increase in serum creatinine from baseline to peak when comparing N-acetylcysteine with placebo (64.5 +/- 91.2 and 38.0 +/- 42.4 mumol/L, respectively; p = .15). Also, there was no attenuation in the increase in serum cystatin C from baseline to peak for N-acetylcysteine compared with placebo (0.45 +/- 0.43 and 0.30 +/- 0.33 mg/L, respectively; p = .40). Likewise, there was no evidence for differences in any other clinical outcome. CONCLUSIONS: In this phase II, randomized, controlled trial, high-dose N-acetylcysteine was no more effective than placebo in attenuating cardiopulmonary bypass-related acute renal failure in high-risk cardiac surgery patients. PMID- 17414731 TI - Predictive value of N-terminal pro-brain natriuretic peptide in severe sepsis and septic shock. AB - OBJECTIVE: The aim of this study was to evaluate the predictive value of N terminal pro-brain natriuretic peptide (NT-proBNP) on mortality in a large, unselected patient population with severe sepsis and septic shock. DESIGN AND SETTING: Prospective observational cohort study about incidence and prognosis of sepsis in 24 intensive care units in Finland (the FINNSEPSIS study). PATIENTS: A total of 254 patients with severe sepsis or septic shock. MEASUREMENTS: After informed consent, the blood tests for NT-proBNP analyses were drawn on the day of admission and 72 hrs thereafter. Patients' demographic data were collected, and intensive care unit and hospital mortality and basic hemodynamic and laboratory data were recorded daily. MAIN RESULTS: NT-proBNP levels at admission were significantly higher in hospital nonsurvivors (median, 7908 pg/mL) compared with survivors (median, 3479 pg/mL; p = .002), and the difference remained after 72 hrs (p = .002). The receiver operating characteristic curves of admission and 72 hr NT-proBNP levels for hospital mortality resulted in area under the curve values of 0.631 (95% confidence interval, 0.549-0.712; p = .002) and 0.648 (95% confidence interval, 0.554-0.741; p = .002), respectively. In logistic regression analyses, NT-proBNP values at 72 hrs after inclusion and Simplified Acute Physiology Score for the first 24 hrs were independent predictors of hospital mortality. Pulmonary artery occlusion pressure (p < .001), plasma creatinine clearance (p = .001), platelet count (p = .03), and positive blood culture (p = .04) had an independent effect on first-day NT-proBNP values, whereas after 72 hrs, only plasma creatinine clearance (p < .001) was significant in linear regression analysis. CONCLUSION: NT-proBNP values are frequently increased in severe sepsis and septic shock. Values are significantly higher in nonsurvivors than survivors. NT-proBNP on day 3 in the intensive care unit is an independent prognostic marker of mortality in severe sepsis. PMID- 17414733 TI - Mentoring and lifelong learning: a critical part of who we are and what we do. PMID- 17414732 TI - Recombinant human activated protein C inhibits local and systemic activation of coagulation without influencing inflammation during Pseudomonas aeruginosa pneumonia in rats. AB - OBJECTIVE: Alveolar fibrin deposition is a hallmark of pneumonia. It has been proposed that recombinant human activated protein C exerts lung-protective effects via anticoagulant and anti-inflammatory pathways. We investigated the role of the protein C system in pneumonia caused by Pseudomonas aeruginosa, the organism that is predominantly involved in ventilator-associated pneumonia. DESIGN: An observational clinical study and a controlled, in vivo laboratory study. SETTING: Multidisciplinary intensive care unit and a research laboratory of a university hospital. PATIENTS AND SUBJECTS: Patients with unilateral ventilator-associated pneumonia and male Sprague-Dawley rats. INTERVENTIONS: Bilateral bronchoalveolar lavage was performed in five patients with unilateral ventilator-associated pneumonia. A total of 62 rats were challenged with intratracheal P. aeruginosa (10 colony-forming units), inducing pneumonia. Rats were randomized to treatment with normal saline, recombinant human activated protein C, heparin, or recombinant tissue plasminogen activator. MEASUREMENTS AND MAIN RESULTS: Patients with pneumonia demonstrated suppressed levels of protein C and activated protein C in bronchoalveolar lavage fluid obtained from the infected site compared with the contralateral uninfected site. Intravenous administration of recombinant human activated protein C in rats with P. aeruginosa pneumonia limited bronchoalveolar generation of thrombin-antithrombin complexes, largely preserving local antithrombin activity. However, recombinant human activated protein C did not have effects on neutrophil influx and activity, expression of pulmonary cytokines, or bacterial clearance. CONCLUSIONS: In patients with ventilator-associated pneumonia, the pulmonary protein C pathway is impaired at the site of infection, and local anticoagulant activity may be insufficient. Recombinant human activated protein C prevents procoagulant changes in the lung; however, it does not seem to alter the pulmonary host defense against P. aeruginosa pneumonia. PMID- 17414734 TI - IKappaB-kinase/nuclear factor-kappaB signaling prevents thermal injury-induced gut damage by inhibiting c-Jun NH2-terminal kinase activation. AB - OBJECTIVE: The molecular mechanism of major burn-induced gut damage is not clear. This study is to determine whether IkappaB-kinase (IKK)/nuclear factor-kappaB signaling in intestinal mucosa maintains gut function through the regulation of the c-Jun NH2-terminal kinase (JNK) and p38 phosphorylation. DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university hospital. SUBJECTS: Thermal injury models in mice. INTERVENTIONS: Conditional intestinal epithelial cell IKKbeta knockout (Vil-Cre/Ikkbeta(F/Delta) mice and control (Ikkbeta(F/Delta) mice were subjected to 30% total body surface area third-degree burn. JNK inhibitor (SP600125) or p38 inhibitor (SB203580) was given to mice immediately after burn injury. MEASUREMENTS AND MAIN RESULTS: Thermal injury induced a significant increase of intestinal permeability, nuclear factor-kappaB DNA-binding activity, phosphorylated JNK, phosphorylated p38, and caspase 3 expression of intestinal mucosa in Vil-Cre/Ikkbeta(F/Delta) mice compared with those of Ikkbeta(F/Delta) mice. BCL-xL and cellular FLICE inhibitory protein, but not GADD45beta (growth arrest and DNA damage-inducing protein beta), cellular inhibitor of apoptosis 1, Bfl-1, or TRAIL, messenger RNA expression was significantly decreased in Vil-Cre/Ikkbeta(F/Delta) mice compared with that of Ikkbeta(F/Delta) mice. SP600125 decreased intestinal permeability and increased phosphorylated p38 and tumor necrosis factor receptor-associated factor 2 expression of intestinal mucosa in Vil-Cre/Ikkbeta(F/Delta) mice. SB203580 treatment enhanced thermal injury-induced gut damage in Vil-Cre/Ikkbeta(F/Delta) mice. CONCLUSIONS: Thermal injury induces nuclear factor-kappaB activation of intestinal mucosa and IKK protects intestinal mucosa from thermal injury-induced gut damage. IKK blocks caspase 3 expression by up-regulating BCL-xL and cellular FLICE inhibitory protein expression. IKK inhibits JNK and p38 but not p44/42 phosphorylation of intestinal mucosa. JNK inhibition increases p38 and tumor necrosis factor receptor-associated factor 2 expression and decreases thermal injury-induced gut damage. Taken together with the enhanced thermal injury induced gut damage by p38 inhibition, we conclude that IKK maintains gut function by inhibiting JNK phosphorylation, which suppresses p38 phosphorylation and induces gut damage. PMID- 17414735 TI - Prediction of short-term and long-term outcomes after cardiac arrest: a prospective multivariate approach combining biochemical, clinical, electrophysiological, and neuropsychological investigations. AB - OBJECTIVE: To determine the prognostic accuracy of biochemical, clinical, electrophysiological, and neuropsychological investigations in predicting outcomes after cardiac arrest. DESIGN: Prospective study. SETTING: Intensive care unit of the Hamburg-Eppendorf University Medical Center, Hamburg, Germany. PATIENTS: A total of 80 patients (mean age, 63.79 +/- 15.85 yrs) after cardiopulmonary resuscitation. INTERVENTIONS: Serial blood samples (days 2-4), clinical examinations (days 2 and 4), sensory-evoked potentials (day 4), and neuropsychological assessments ( 0.44). CONCLUSIONS: Parallel imaging performed in a 3-T MR system improves hepatic MRA both quantitatively and qualitatively. PMID- 17414750 TI - Hepatic venous congestion after right-lobe living-donor liver transplantation: the added value of delayed-phase imaging on CT. AB - PURPOSE: To describe and determine the additional value of delayed-phase imaging of hepatic venous congestion after living-donor liver transplantation. MATERIALS AND METHODS: Twenty-eight patients who had surgical ligation of the middle hepatic vein (HV) in living-donor liver transplantation underwent 3-phase computed tomography scans. Two radiologists analyzed in consensus the presence and pattern of the hepatic attenuation difference and the opacification of the HV in the congested areas of the liver during each phase of the initial and follow up computed tomography scanning. The imaging findings were correlated with the serum bilirubin level. RESULTS: Opacification of the HV was observed more frequently in 22 (92%) of 24 hyperattenuating areas on delayed-phase (DP) scans than in 2 (50%) of 4 hypoattenuating areas in the congested areas of the liver. Patients with persistent hypoattenuatation in the congested areas on all phases (14%) showed significantly persistent hyperbilirubinemia after postoperative 4 weeks and showed a higher mortality rate (50%) than did the other patients with hyperattenuation on DP scans. CONCLUSIONS: A hypoattenuating area of the liver during DP scans indicates severe hepatic congestion and is correlated with hyperbilirubinemia and a high mortality rate. PMID- 17414751 TI - Poorly versus moderately differentiated hepatocellular carcinoma: vascularity assessment by computed tomographic hepatic angiography in correlation with histologically counted number of unpaired arteries. AB - PURPOSE: To determine the vascularity of moderately and poorly differentiated hepatocellular carcinoma (mHCC and pHCC, respectively) as observed on and depicted by computed tomography during hepatic angiography and to perform pathological correlation. MATERIALS AND METHODS: Eighty-seven consecutive patients with 89 hepatocellular carcinomas (61 mHCCs and 28 pHCCs) were surgically resected in our hospital. The degree of contrast enhancement on computed tomography during hepatic angiography of the tumors was classified into high attenuation (H), isoattenuation (I), and low attenuation (L). We also examined hepatocellular carcinomas measuring less than 4 cm in diameter. Pathologically, the number of unpaired arteries in the tumors was determined (x200 magnification). RESULTS: The number of mHCC and pHCC in each degree of enhancement (H/I/L) was 59:1:1 and 19:6:3, respectively. The number of mHCC and pHCC measuring less than 4 cm without portal invasion was 48 and 15, respectively; the number of these tumors in each degree of enhancement (H/I/L) was 47:1:0 and 11:3:1, respectively. The mean number of unpaired arteries was 8.9 +/- 4.4 in mHCC and 5.2 +/- 4.3 in pHCC, respectively. All results were statistically significant (P < 0.01). CONCLUSIONS: Our results indicated that the arterial blood supply of pHCC was lower than that of mHCC. PMID- 17414752 TI - Free-breathing motion compensation using template matching: a technique allowing for tracer kinetic modeling of liver metastases. AB - Modeling tracer kinetics from dynamic magnetic resonance imaging (dMRI) to understand microvascular characteristics typically requires acquisitions longer than 1 breath-hold. This has limited the application of dMRI in assessment of the upper abdomen. Here we present a template-based motion correction strategy for dMRI of liver metastases based on the correlation coefficient (CC), originally developed for tracking coronary arteries. This postprocessing method allows patient free breathing during sagittal dMRI acquisition and allows a more precise parametric mapping using tracer kinetic models. In a study of 6 subjects, a 64 x 64 template was accurately tracked retrospectively with mean CC = 0.72 +/- 0.07. Mean superior-inferior displacement tracked was 1.82 +/- 1.20 pixels, whereas mean anterior-posterior displacement was 7.72 +/- 4.58 pixels. Application of the CC method significantly improved the global fit (chi2) of a tracer kinetic model throughout tumor regions. Therefore, use of the CC postprocessing method for dMRI scans can improve the precision of dMRI tracer kinetic models. PMID- 17414753 TI - Prediction of viable tumor in hepatocellular carcinoma treated with transcatheter arterial chemoembolization: usefulness of attenuation value measurement at quadruple-phase helical computed tomography. AB - OBJECTIVE: To assess the usefulness of attenuation value measurement at quadruple phase helical computed tomography (CT) for predicting viable tumor in hepatocellular carcinoma (HCC) treated with transcatheter arterial chemoembolization (TACE). METHODS: Thirty-two patients who had an iodized-oil defect area (IODA) in HCCs treated with TACE were included in our study; these patients were divided into group 1 (n = 21) with viable tumor and group 2 (n = 11) without viable tumor in the IODA. All the patients underwent quadruple-phase helical CT (unenhanced and contrast-enhanced hepatic arterial, portal venous and equilibrium phases) before and after TACE. The attenuation difference of the IODA between unenhanced and each contrast-enhanced phase was measured, and the attenuation degree of the IODA relative to the hepatic parenchyma were subjectively assessed and then compared between the 2 groups. RESULTS: The mean attenuation differences of the IODAs were 28.8, 35.9, and 25.6 Hounsfield unit (HU) in group 1 and 0.4, 1.9, and 2.0 HU in group 2 at the hepatic arterial, portal venous, and equilibrium phases, respectively, with statistically significant difference for each phase (P < 0.05). The IODAs had attenuation difference of more than 20 HU on at least 1 contrast-enhanced phase in group 1 and less than 5 HU at all contrast-enhanced phases in group 2. For the attenuation degree of IODAs relative to the hepatic parenchyma, 12 patients (57%) showed hyperattenuation at the hepatic arterial phase, and remaining nine (43%) at the hepatic arterial phase and all patients at the portal venous and equilibrium phases showed isoattenuation or hypoattenuation in group 1. In group 2, all the patients showed hypoattenuation at all the 3 phases. CONCLUSIONS: The presence of viable tumor of the IODA in HCC treated with TACE can be precisely assessed by measuring attenuation values, strongly suggesting viable tumor when the attenuation difference is more than 20 HU on at least 1 contrast-enhanced phase at quadruple-phase helical CT. PMID- 17414754 TI - Gastric true leiomyoma: computed tomographic findings and pathological correlation. AB - OBJECTIVE: This study describes the computed tomographic (CT) findings of true leiomyomas in the stomach. METHODS: Eleven patients with histopathologically proven gastric true leiomyomas were finally enrolled. All cases were CD117 (c kit) negative on immunohistochemical study. We retrospectively reviewed the CT findings of gastric true leiomyomas for the following considerations: location, morphological features (size, contour, tumor growth pattern, and enhancement pattern), and ancillary findings (ulceration and calcification). RESULTS: All leiomyomas were in the cardia, with an average tumor size of 36 mm (range, 13-47 mm). Computed tomography scans revealed the morphological features as follows: tumor growth pattern: intraluminal mass (n = 10) and extraluminal masses (n = 1); contour: lobulated margin (n = 7) or smooth margin (n = 4). All lesions showed homogeneous contrast enhancement. Most of the tumors showed lower enhancement than those in the liver (n = 10). One case showed ulceration but no calcification. CONCLUSIONS: In conclusion, gastric leiomyomas are mainly located in the cardia and usually appeared as homogenous low attenuated masses on computed tomography. PMID- 17414755 TI - Determination of 3-dimensional zonal renal volumes using contrast-enhanced computed tomography. AB - OBJECTIVE: To determine the accuracy with which total and cortical renal volumes may be measured in vivo with contrast-enhanced computed tomography (CT) in an animal model. METHODS: Seven female Yorkshire pigs were scanned in vivo using both a 64-channel multidetector row computed tomography (MDCT) scanner (Siemens Medical Solutions, Erlangen, Germany) and a C-arm CT scanner (Siemens AXIOM). Kidneys were scanned in corticomedullary and nephrographic phases after contrast injection. Three-dimensional volumetric analysis of CT data was performed to determine renal cortical and overall renal volumes, and renal lengths. We measured renal weights, water displacement volumes, and overall and cortical renal volumes of resected kidneys. RESULTS: Overall and cortical renal volumes by CT showed excellent correlation (R2 values varying from 0.77 to 0.92) with anatomic measurements of renal volume. There was poor correlation of anatomic renal volumes with renal lengths by CT. CONCLUSIONS: Three-dimensional analysis of contrast-enhanced CT provides accurate measurement of overall and cortical renal volumes in vivo. PMID- 17414756 TI - Interferometer-based phase-contrast X-ray computed tomography of colon cancer specimens: comparative study with 4.74-T magnetic resonance imaging and optical microscopy. AB - OBJECTIVE: Phase-contrast X-ray computed tomography (PCCT) with an interferometer can reveal the inner soft tissue structures of biological objects without contrast agent, and the image quality is thought to resemble that of magnetic resonance imaging (MRI). Comparative study among PCCT, MRI, and optical microscopy was undertaken. METHODS: Three formalin-fixed colon cancer specimens from nude mice were imaged both by PCCT with a reconstructed volumetric resolution of (0.018)3 mm3 and 4.74-T MRI with that of (0.075)3 mm3. RESULTS: Phase-contrast X-ray computed tomography with an interferometer clearly demonstrated the inner structures of colon cancer masses, such as cancer, necrosis, surrounding tumor vessels, and skin, in a similar way to low-magnified optical microscopic images and had approximately 4.0-fold higher signal-to-noise ratio than MRI. CONCLUSIONS: With formalin-fixed biological samples, PCCT exhibited higher image quality than MRI and was thought to be suitable for detailed imaging of soft tissue with high volumetric resolution. PMID- 17414757 TI - Comparison of 2 different protocols for ingestion of low-attenuating oral contrast agent for multidetector computed tomography of the abdomen. AB - OBJECTIVE: To evaluate 2 different oral contrast application protocols concerning degree of intestinal contrast filling and distension in multidetector computed tomography. METHODS: Examinations of 260 patients were retrospectively analyzed. Group 1 (n = 205) was prepared with 1000 mL of water with 28 g of dissolved mannitol which was ingested for 1 hour; group 2 (n = 55) was prepared with 2000 mL of water containing 56 g of mannitol which was ingested for 2 hours. Small intestine was divided into 4 quadrants; colon was divided into 4 segments. Contrast filling and distension of bowel loops were graded with a 3-point scale and compared using chi2 testing. RESULTS: Besides the right upper quadrant, no significant differences in contrast filling and distention were found for small bowel segments. Colonic segments were significantly better contrast filled and distended in group 2 (0.001 < P < 0.025). CONCLUSIONS: A higher amount of water with 28 g/L of mannitol and a prolonged ingestion of oral contrast media result in significantly better contrast filling and bowel distension in the ileocecal region, the colon, and the rectum. PMID- 17414758 TI - [F-18]fluorodeoxyglucose positron emission tomography and positron emission tomography: computed tomography in recurrent and metastatic cholangiocarcinoma. AB - OBJECTIVES: We retrospectively assessed the diagnostic utility of dedicated positron emission tomography (PET) and hybrid PET-computed tomography (CT) scans with [F-18]fluorodeoxyglucose (FDG) in the imaging evaluation of patients with known or suspected recurrent and metastatic cholangiocarcinoma. METHODS: The study group included 24 patients (13 males and 11 females; age range, 34-75 years) with known or suspected recurrent and metastatic cholangiocarcinoma. We performed 8 dedicated PET scans (Siemens 953/A, Knoxville, Tenn) in 8 patients and 24 hybrid PET-CT scans (Siemens Biograph, Knoxville, Tenn) in 16 patients. Four patients underwent both pretreatment and posttreatment scans. Nonenhanced CT transmission scans were obtained for attenuation correction after administration of oral contrast material. PET images were obtained 60 minutes after the intravenous administration of 15 mCi (555 MBq) FDG. Prior treatments included surgery alone in 12 patients, surgery and chemotherapy in 6 patients, and surgery and combined chemoradiation therapy in 6 patients. Diagnostic validation was conducted through clinical and radiologic follow-up (2 months to 8 years). RESULTS: PET and CT were concordant in 18 patients. PET-CT correctly localized a hypermetabolic metastatic lesion in the anterior subdiaphragmatic fat instead of within the liver and was falsely negative in intrahepatic infiltrating type cholangiocarcinoma. PET was discordant with CT in 6 patients. PET was negative in an enlarged right cardiophrenic lymph node on CT, which remained stable for 1 year. In 1 patient, PET-CT scan showed hypermetabolic peritoneal disease in the right paracolic gutter without definite corresponding structural abnormalities, which was subsequently confirmed on a follow-up PET-CT scan performed 6 months after the initial study, at which time peritoneal nodular thickening was evident on concurrent CT. PET-CT documented the progression of locally recurrent and metastatic disease in another patient based on interval appearance of several new hypermetabolic lesions and significant increase in the standardized uptake values of the known lesions despite little interval change in the size and morphologic character of lesions on concurrent CT. It was also helpful in excluding metabolically active disease in patients with contrast enhancement at either surgical margin of hepatic resection site or focally within hepatic parenchyma and in an osseous lesion. Overall, based on the clinically relevant patient basis for detection of recurrent and metastatic cholangiocarcinoma, the sensitivity and specificity of PET (alone and combined with CT) were 94% and 100% and, for CT alone, were 82% and 43%, respectively. CONCLUSIONS: FDG PET and PET-CT are useful in the imaging evaluation of patients with cholangiocarcinoma (except for infiltrating type) for detection of recurrent and metastatic disease and for assessment of treatment response. In particular, the combined structural and metabolic information of PET-CT enhances the diagnostic confidence in lesion characterization. PMID- 17414759 TI - Magnetic resonance imaging of endometrial cancer and clear cell cancer. AB - OBJECTIVE: We attempt to clarify specific imaging findings useful for differential diagnosis between endometrioid cancer and clear cell cancer. METHODS: We examined magnetic resonance imaging features on 11 endometrioid cancer and 9 clear cell cancer patients. We compared them, focusing on the locularity, characteristic and growth pattern of solid components, and association with endometrial diseases. RESULTS: Endometrioid cancer had a predominantly multilocular appearance, but clear cell cancer showed almost equally multilocular and unilocular appearances. For the growth pattern, endometrioid cancer in 6 patients demonstrated a "centric pattern" and an "eccentric pattern" in 4, whereas clear cell cancer showed predominantly an eccentric pattern. An internal slit in the solid component was seen in 4 endometrioid cancer patients but not seen in any of the patients with clear cell cancer. CONCLUSIONS: The magnetic resonance imaging appearance of the internal slit in solid components and the presence of endometrial disease are key factors in differentiating endometrioid cancer from clear cell cancer, although lesion locularity and growth pattern are not specific in both. PMID- 17414760 TI - Value of dynamic enhanced magnetic resonance imaging for distinguishing between ovarian fibroma and subserous uterine leiomyoma. AB - PURPOSE: The purpose of this study was to assess the accuracy of magnetic resonance imaging (MRI), particularly, dynamic MRI, in distinguishing ovarian fibromas from subserous uterine leiomyomas. MATERIAL AND METHODS: Fifteen ovarian fibromas and 15 subserous uterine leiomyomas were retrospectively reviewed. All MR examinations included dynamic contrast-enhanced (DCE) sequences. Morphological criteria (size, T1 and T2 signals, ovarian tissue, associated uterine leiomyoma, and pelvic fluid), arterial vessels, time-intensity curves (maximal enhancement and signal intensity at 30, 60, and 90 seconds), and signal intensity on delayed T1-weighted images were compared between the 2 groups. RESULTS: No significant difference in morphological criteria was noted between the 2 types of masses. Visualization of arterial vessels was more frequent in uterine leiomyomas than in ovarian fibromas (P= 0.002). The DCE MR enhancement rate was higher for uterine leiomyomas than for ovarian fibromas in terms of both maximal enhancement (P < 0.001) and enhancement rate at 30 (P = 0.009), 60 (P = 0.007), and 90 seconds (P = 0.0009). On delayed T1 postcontrast sequence, no statistical difference exists between signal intensity of ovarian fibromas and uterine leiomyomas. CONCLUSION: Our findings suggest that DCE MRI can distinguish ovarian fibromas from uterine leiomyomas and should be used if sonography fails to show the origin of a pelvic mass. PMID- 17414761 TI - Magnetic resonance fistulography for the demonstration of anovaginal fistula: an alternative imaging technique? AB - Anovaginal fistulae (AVF) are frequently seen in patients with inflammatory bowel disease, especially in Crohn disease with active colonic inflammation. Herein, we report a 21-year-old woman with Crohn disease suffering from vaginal discharge and anal pain. Although clinical presentation was very suggestive of AVF, physical examination and colonoscopy were inconclusive. We used an alternative technique and performed magnetic resonance fistulography by applying rectal contrast for the demonstration of AVF. PMID- 17414762 TI - Superficial endometrial spread of squamous cell cervical carcinoma: a diagnostic challenge at magnetic resonance imaging. AB - The magnetic resonance imaging findings in a case of superficial endometrial spread of squamous cervical carcinoma are presented. Early enhancement of a relatively small cervical mass was associated with a fluid-distended endometrial cavity. An irregular layer of early-enhancing tumor was seen lining the endometrial cavity. Histopathologic examination confirmed the presence of an invasive squamous cell carcinoma of the cervix that superficially spread into the lower uterine segment and endometrial cavity, replacing most of the endometrial lining as a layer of tumor. The tumor also deeply invaded the myometrium in a permeative fashion throughout the bulk of the uterine body. Failure to appreciate the cervical abnormality might have resulted in an imaging diagnosis of endometrial carcinoma. We report this challenging case to broaden the described spectrum of magnetic resonance imaging findings in cervical carcinoma, because recognition of this unusual pattern of tumor spread may facilitate correct radiological interpretation. PMID- 17414763 TI - Magnetic resonance imaging of the ischiorectal fossa: spectrum of disease. AB - OBJECTIVE: Pelvic magnetic resonance imaging (MRI) studies are widely used to assess the rectum, anal canal, and their environment. The purpose of this article is to review the current role of MRI in the evaluation of diseases involving the ischiorectal fossa and their imaging features. CONCLUSIONS: The radiologist plays an essential role in the evaluation of some conditions typically located in this space and of other conditions that occasionally involve this area or invade the ischioanal space. The accurate assessment of these diseases that MRI provides further cements its role as the primary technique for the evaluation of pelvic pathology. PMID- 17414764 TI - Use of 40-detector row computed tomography before catheter coronary angiography to select early conservative versus early invasive treatment for patients with low-risk acute coronary syndrome. AB - OBJECTIVE: Using catheter coronary angiography (CAG) as reference standard, we examined the agreement of 40-detector row computed tomography (MDCT) in triaging patients into the 2 controversial strategies of managing low-risk acute coronary syndrome (ACS). METHODS: Seventy-eight patients with low-risk ACS received both MDCT and CAG. Early invasive strategy was assigned for the patient if there was significant stenosis (> or =50% diameter stenosis) in any of the coronary artery segments with diameter larger than 1.5 mm. The results of MDCT were compared with the CAG for agreement. RESULTS: The overall agreement of the early conservative/early invasive strategy assignment was 92.3%, with kappa value of 0.82 between MDCT and CAG. Only 1 patient needing early invasive strategy was missed by MDCT. CONCLUSION: Forty-detector row computed tomography is reliable in triaging patients into the 2 strategies of managing low-risk ACS. PMID- 17414765 TI - Optimization of contrast material administration for electrocardiogram-gated computed tomographic angiography of the chest. AB - OBJECTIVE: Electrocardiogram-gated computed tomographic angiography is increasingly used in the differential diagnosis of acute chest pain. We studied the optimal timing of contrast material injection using a test bolus and a bolus tracking technique. MATERIALS AND METHODS: Thirty patients were prospectively included in the study. Volume and flow of high concentration contrast material were adapted to body weight. The scan delay was determined using either a test bolus or a bolus-tracking technique. Attenuation profiles of the different vascular districts were measured to evaluate the timing techniques. RESULTS: In all the patients except for one, an adequate and homogeneous contrast enhancement of more than 200 Hounsfield units (HU) was achieved (285 +/- 45 HU) in the different vascular districts. The pulmonary transit time in the test bolus group was 7 seconds (range, 4-11 seconds). Differences and variability of pulmonary and aortic enhancement were small in both groups (13 +/- 48 HU vs -9 +/- 21 HU), with differences of less than 70 HU over the craniocaudal range and very small intraindividual differences between pulmonary attenuation and systemic attenuation. CONCLUSIONS: Contrast administration regimens for electrocardiogramgated computed tomographic angiography of the chest can be optimized using the bolus-tracking method in the ascending aorta, with a short delay after trigger. Body weight adaptation of volume and injection rate of the contrast material results in a reliable simultaneous opacification of the pulmonary and systemic vasculature. PMID- 17414766 TI - Coronary angiography by 64-detector row computed tomography using low dose of contrast material with saline chaser: influence of total injection volume on vessel attenuation. AB - OBJECTIVE: To assess the influence of total injection volume on thoracic great vessels and coronary arteries enhancement in 64-detector row computed tomography (CT) coronary angiography using low dose of contrast material. METHODS: Sixty patients underwent cardiac CT (64 x 0.5 mm, 0.4 rot/s) using 40 mL of contrast material (350 mg of Iodine per milliliter) in 30 patients and 50 mL in 30 patients. Computed tomography densities (Hounsfield units) in ascending aorta, descending aorta, and main pulmonary artery were measured at every second with the time of CT data acquisition recorded in each reconstructed image. Computed tomography densities of proximal and distal coronary arteries were also measured. Differences in CT densities between 40 and 50 mL contrast material were assessed with the Student t test. In addition, the relation between the injection volume (mL) of contrast material per kilogram body weight and contrast enhancement in coronary arteries was studied. RESULTS: The average attenuations in the ascending and descending aorta and coronary arteries were significantly lower in 40-mL group than in 50-mL group (<0.05). In addition, the average attenuations in the pulmonary artery were significantly lower in 40-mL group than 50-mL group (<0.01). Every patient with the total injection volume of more than 0.9 mL/kg body weight showed a contrast enhancement more than 250 Hounsfield units. CONCLUSIONS: The reduction of total injection volume lowered the enhancement of thoracic great vessels and coronary arteries in 64-detector row cardiac CT. The injection volume of at least 0.9 mL/kg body weight was necessary for a steady contrast enhancement in coronary arteries. PMID- 17414767 TI - Patient-specific contrast injection protocols for cardiovascular multidetector row computed tomography. AB - OBJECTIVE: To develop patient-specific contrast injections for uniform enhancement of cardiovascular multidetector row computed tomography (MDCT) images. METHODS: Sixty-two patients were imaged using electrocardiogram (ECG) gated spiral MDCT. Thirty patients (group 1) received a uniphasic injection; the remaining 32 patients (group 2) received patient-specific multiphasic injections. For group 2 patients, the vasculature between injection and imaging sites was considered a "gray box" whose transfer function was determined from a test bolus injection and the resulting enhancement in the left side of the heart. This transfer function was used to determine the injection necessary to achieve 250 Hounsfield units in the left side of the heart. Intraindividual and interindividual variation of enhancement were determined for both groups. Superior vena cava (SVC) artifacts were graded on a 4-point scale. RESULTS: The measured indices of intraindividual variation were significantly smaller in group 2 than in group 1 (P < 0.05), indicating improved uniformity with patient specific injections. The interindividual variation of mean enhancement in group 2 was smaller than in group 1, but the difference was not significant. The severity of SVC artifacts was significantly reduced (P < 0.05) for thinner patients (<83 kg) in group 2 compared with similar patients in group 1. CONCLUSIONS: Patient specific multiphasic contrast injections yielded more uniform enhancement in the left side of the heart on MDCT images with reduced intraindividual variation of enhancement compared with standard uniphasic injections. Patient-specific injections also reduced SVC artifacts in patients <83 kg. PMID- 17414768 TI - Assessment of myocardial viability in a reperfused porcine model: evaluation of different MSCT contrast protocols in acute and subacute infarct stages in comparison with MRI. AB - OBJECTIVE: To assess myocardial viability in acute and subacute infarcts using different multislice spiral computed tomography contrast protocols with magnetic resonance imaging (MRI) correlation. METHODS: Seven pigs were studied with 64 multislice spiral computed tomography and MRI (1.5 T) at a median of 1 and 21 days after temporary occlusion of the second diagonal branch. Computed tomography was performed at 3, 5, 10, and 15 minutes after injection of contrast medium. Contrast agent was applied either as a bolus (protocol 1; n = 7 for the first; n = 5 for the second scan) or as a bolus plus 30 mL of subsequent 0.1 mL/s low-flow (protocol 2; n = 7 for the first; n = 6 for the second scan). Finally, histological sections were obtained. Volumes of infarcted myocardium were assessed as the percentage of the left ventricle. Computed tomography attenuation values were obtained, and image quality was assessed. RESULTS: When compared with protocol 1, protocol 2 provided greater Hounsfield unit attenuation difference between viable and nonviable myocardium at 5, 10, and 15 minutes (P = 0.19; 0.003; 0.0006) and an additional significant contrast between nonviable myocardium and ventricular blood at 3 and 5 minutes (P < 0.001). Image quality was rated significantly higher with the use of protocol 2 at 5, 10, and 15 minutes (P < or = 0.027) and for all time points use of protocol 2 resulted in improved correlation of acute and subacute infarct size with MRI. CONCLUSIONS: Good correlation of infarct zones with MRI was achieved for both acute and subacute infarcts. With the use of a bolus/low-flow protocol, image quality was substantially improved by means of a higher tissue contrast. PMID- 17414769 TI - Nonenhanced magnetic resonance lymphoductography: visualization of lymphatic system of the trunk on 3-dimensional heavily T2-weighted image with 2-dimensional prospective acquisition and correction. AB - The objective of this study was to evaluate the usefulness of the nonenhanced 3 dimensional heavily T2-weighted images obtained with 2-dimensional prospective acquisition and correction in visualization of the lymphatic system of the trunk. The thoracic duct, cisterna chyli, and lumbar lymphatics were well shown in 9 healthy volunteers. This nonenhanced magnetic resonance lymphoductography is thought to be a suitable method in visualization of the lymphatic system of the trunk. PMID- 17414770 TI - Magnetic resonance imaging of lymphatic vessels without image subtraction: a practicable imaging method for routine clinical practice? AB - OBJECTIVE: To assess the feasibility of a time-efficient, high-resolution magnetic resonance lymphangiography (HR MRL) protocol without image subtraction for the detection of lymphatic vessels in patients with primary and secondary lymphedema. METHODS: Three consecutive patients with lymphedema of the lower extremities (2 primary bilateral, 1 secondary unilateral) underwent HR MRL without image subtraction. An amount of 9 mL of gadodiamide and 1 mL of mepivacaine hydrochloride 1% were subdivided into 5 portions and injected intracutaneously into the dorsal aspect of each foot outside the scanner before image acquisition. Magnetic resonance imaging was performed with a 1.5-T system equipped with high-performance gradients. For HR MRL, a 3-dimensional, spoiled gradient-echo sequence (Volumetric Interpolated Breath-hold Examination) was used. The extent and distribution of the lymphedema was evaluated using a heavily T2-weighted, 3-dimensional turbo-spin echo sequence. RESULTS: The HR MRL bilaterally detected the inguinal lymph nodes and the lymphatic vessels in the lower and upper leg in the 2 patients with primary lymphedema. In the patient with left-sided secondary lymphedema, the inguinal lymph nodes and the lymphatic vessels in the lower and upper leg were depicted on the right side. The diameter of the displayed lymphatic vessels varied between 1 and 5 mm. Three-dimensional, maximum-intensity projection images of different angles of view provided detailed outlining of the lymphatic vessels and differentiation from veins, which showed a lower signal intensity. CONCLUSION: The HR MRL without image subtraction is safe, technically feasible, and has the potential to become a diagnostic imaging tool in daily clinical practice because of its time efficiency. PMID- 17414771 TI - Dynamic multislice computed tomography findings for parotid gland tumors. AB - OBJECTIVE: Our aim was to research the enhancement features of parotid gland masses in detail and characterize if the masses were Warthin tumors, adenomas, or malignant tumors. METHODS: The prospective study included 25 parotid tumors in 21 patients. Neck computed tomography (CT) was performed using a multislice CT unit. A full-neck CT examination was done at 30 seconds after completion of contrast injection, and then tumor-level images were obtained at 90 seconds and at 5 and 25 minutes. Computed tomography number (lesion density in Hounsfield units) was determined at each phase, and differences within and among tumor groups were statistically analyzed. Diagnoses were confirmed by histopathology. RESULTS: There were 11 Warthin tumors, 8 pleomorphic adenomas, 5 malignant tumors, and 1 basal cell adenoma. Ten Warthin tumors showed rapid contrast enhancement at 30 seconds and rapid reduction of enhancement from the first to the fourth phase. The basal cell adenoma showed also a peak enhancement at 30 seconds. Seven pleomorphic adenomas showed increased enhancement through the first 3 phases. Four malignant tumors showed peak enhancement at 90 seconds. Statistically significant differences within and among tumor groups were determined. CONCLUSIONS: The data suggest that peak tumor enhancement at 30 and 90 seconds, respectively, might identify Warthin and malignant tumors. Increased enhancement through all phases might be an indicator for diagnosing pleomorphic adenomas. PMID- 17414772 TI - Gliomatosis peritonei associated with immature ovarian teratoma: a mimicker of peritoneal dissemination of malignant diseases. AB - A rare case of gliomatosis peritonei associated with immature ovarian teratoma was reported. The computed tomographic findings of gliomatosis peritonei were similar to the classic findings of peritoneal dissemination of malignant tumors. To avoid overestimating the clinical stage of ovarian tumors, radiologists should be aware of this rare condition related to teratomas. PMID- 17414773 TI - Quantitative microcomputed tomography assessment of intratrabecular, intertrabecular, and cortical bone architecture in a rat model of severe renal osteodystrophy. AB - OBJECTIVE: To determine the effects of renal osteodystrophy (ROD) on bone microarchitecture in growing rats. METHODS: A total of 24 rats underwent 5/6 nephrectomy (NX) and were fed a high-phosphorus diet to induce ROD; another 6 underwent sham NX. In vitro microcomputed tomography images (GEMS, London, Ontario, Canada) were obtained in the femoral metaphysis and midshaft. RESULTS: Trabecular and cortical bone volume/total volume (BV/TV) were significantly lower in NX specimens because of pores within the trabeculae and along the endosteal surface. Topological analysis using component labeling in 3-dimensions verified that trabecular pores connected to the marrow space. After the trabecular pores were filled using a morphological filter, trabecular thickness was significantly increased in NX. In contrast, cortical thickness was significantly decreased in NX compared with controls; however, after filling the endocortical pores, thickness did not differ. CONCLUSIONS: The ROD resulted in decreased cortical and trabecular BV/TV, increased porosity, and increased trabecular thickness. Advanced image processing algorithms demonstrated the effects of cortical and trabecular porosity on BV/TV and structure in ROD. PMID- 17414774 TI - Body mass index in ambulatory cerebral palsy patients. AB - Malnutrition is a common problem in children with cerebral palsy. Although malnutrition is often recognized in patients with severe cerebral palsy, it can be unrecognized in less severely affected patients. The consequences of malnutrition are serious, and include decreased muscle strength, poor immune status, and depressed cerebral functioning. Low body mass index has been used as a marker for malnutrition. The purpose of this study was to determine which patients in an ambulatory cerebral palsy patient population were at risk for low body mass index. A retrospective chart review was performed on 75 patients. Age, sex, height, weight, type of cerebral palsy, and functional status [gross motor functional classification system (GMFCS) level] was recorded from the chart. Descriptive statistics with bivariate and multivariate regression analyses were performed. Thirty-eight boys and 37 girls with an average age of 8.11 years were included in the study. Unique to our patient population, all cerebral palsy patients were independent ambulators. Patients with quadriplegic cerebral palsy had a significantly lower body mass index than those with diplegic and hemiplegic cerebral palsy. Patients with a GMFCS III had significantly lower body mass index than those with GMFCS I and II. When multivariate regression analysis to control for age and sex was performed, low body mass index remained associated with quadriplegic cerebral palsy and GMFCS III. Malnutrition is a common health problem in patients with cerebral palsy, leading to significant morbidity in multiple organ systems. We found that in an ambulatory cerebral palsy population, patients with lower functional status or quadriplegia had significantly lower body mass index, suggesting that even highly functioning ambulatory cerebral palsy patients are at risk for malnutrition. PMID- 17414775 TI - Epiphyseal separations in spastic cerebral palsy. AB - Metaphyseal and diaphyseal fractures have been reported in children with severe spastic cerebral palsy, but there is no reference to epiphyseal separations in these patients. We report nine epiphyseal separations involving the distal femur and proximal humerus in four severely affected children with spastic quadriplegic cerebral palsy. The clinico-radiological features confirmed the cause to be scurvy. These epiphyseal separations have a good prognosis unlike the physeal injuries reported in children with myelodysplasia. Treatment with vitamin C and splintage resulted in rapid healing with excellent remodeling. Scurvy should be considered a potential cause for an epiphyseal slip in a child with severe spastic cerebral palsy. Routine dietary supplementation of vitamin C is recommended for these malnourished and nonambulant children. PMID- 17414776 TI - Evaluating upper body movements during gait in healthy children and children with diplegic cerebral palsy. AB - Movements of the lower limbs during gait have been analysed extensively whereas data on upper body movements are scarce. The aim of this study was to evaluate upper body movements during gait in nine healthy children and 10 children with diplegic cerebral palsy. Children were investigated using a full-body marker set to calculate the upper body kinematics of trunk and arms. When the healthy children were compared with the children with cerebral palsy, the latter compensated more for their gait deviations and were less stable. This was expressed by their greater variability in arm movements and increased movements at the thorax. The thorax showed an increased forward tilt with greater range of motion over the gait cycle. The shoulders were more abducted with increased elbow flexion. Gait analysis with the full-body marker set has offered prospects for a better understanding of compensatory mechanisms for the pathological gait pattern in children with diplegic cerebral palsy. PMID- 17414777 TI - Proximal femoral resection-interposition arthroplasty in cerebral palsy. AB - Hip dislocation is common in children with total body cerebral palsy. It is associated with pain, difficulty in seating and problems with perineal hygiene. We present a retrospective study of 15 patients who had undergone 21 proximal femoral resection-interposition arthroplasties between 1990 and 2005. The mean age at the time of surgery was 16.2 years (range 11-26 years). Mean follow-up was 3.4 years (range 1-7 years). Ninety percent of patients had pain relief. Perineal hygiene and sitting was improved in all cases. A lateral femoral head defect was noted in 90% of hips and was associated with pain, and significant degenerative changes. Twelve hips had heterotopic ossification (11 type-1, one type-2 and 0 type-3). No major complications were seen. We would submit that proximal femoral resection is a good salvage procedure for the painful, dislocated hip, with an excellent chance of resolving symptoms. The finding of a lateral femoral head defect on the radiograph is associated with significant degenerative changes. PMID- 17414778 TI - Management of long toe flexor spasticity in the equinus foot in cerebral palsy. AB - Toe flexor tightness becomes apparent in the plantigrade attitude achieved by isolated tendoachilles lengthening in cerebral palsy. This frequently persists, resulting in abnormal weight-bearing on the tips of the toes. To address this problem, at the time of limited tendoachilles lengthening, we performed Z lengthenings of the flexor hallucis longus and the flexor digitorum longus tendons above the medial malleolus, preserving the pulley mechanism. From January 1998 to October 2001, 41 children (74 feet) with cerebral palsy underwent lengthening of the tendoachilles and long toe flexor tendons. The mean age at surgery was 7 years. The correction achieved was successfully maintained long term, permitting a normal heel-metatarsal head weight-bearing pattern. Lengthening of the long toe flexors, in conjuction with tendoachilles lengthening, is an effective means of achieving a balanced foot in cerebral palsy. PMID- 17414779 TI - The lack of evidence of the effect of selective ultrasound screening on the incidence of late developmental dysplasia of the hip in the Greater Glasgow Region. AB - The aim of our study was to establish whether the use of ultrasound to screen babies with risk factors has reduced the number of children presenting late with developmental dysplasia of the hip. Screening based on selection by risk factors was introduced in 1997. The average annual incidence from 1992 to 1996 was 0.84 and from 1997 to 2001 was 0.57 per 1000 live births. This decline in incidence of late developmental dysplasia of the hip was not significant (P=0.088). Sixty-four children (82%) had nothing that could be perceived as a risk factor for the condition. Our study presents population-based findings and it is clear that selective screening is not working in our region. It is unlikely that there will be one solution that could be applied throughout the United Kingdom given the diversity in the access to healthcare facilities. Clear leadership and accountability are required if we are to make any progress towards abolishing this condition that causes a significant morbidity in young adults. PMID- 17414780 TI - Ultrasound screening in developmental hip dysplasia: do all scanned hips need to be followed up? AB - Many ways of detecting hip instability in the newborn infant exist, including a history of risk factors, clinical examination and ultrasound. We investigated our practice of 'at risk' screening using subjective, dynamic and static, ultrasound, followed by radiographic evaluation at 12 months. We found that the average age at presentation was 71 days, with the most common reason for referral being a clicking hip. None of the hips deemed stable under review deteriorated, and all those considered normal at first visit had a normal radiograph at 12 months. Abnormal radiographs were found in all the other groups. PMID- 17414781 TI - Role of preoperative arthrography in early Perthes disease as a decision-making tool. Is it really necessary? AB - Arthrograms are commonly done in conjunction with an examination under anaesthesia (EUA), before any surgical intervention in Perthes disease. This usually is done as an independent day case procedure in the radiology department. The aim of this study is to confirm that the affected hip meets the prerequisites of a containment procedure. This study was carried out to investigate whether arthrogram findings ever altered the preoperative clinical decision of proposed osteotomy. One hundred and seven cases were reviewed retrospectively. Forty patients were excluded. Decision was not changed in 62 out of 67 hips, after the procedure. Five patients required different management following arthrogram and this was based mainly on EUA findings. EUA under image intensification helps the surgeon to decide the angle of varus osteotomy, required to contain the femoral head in the acetabulum and confirms an adequate range of abduction to undertake the procedure. This quick assessment can be done at the time of surgery under the same anaesthesia without the need for arthrography. Our findings question the role of arthrography as a routine before surgical intervention in Perthes disease and suggest that it may represent an unnecessary cost. We now reserve arthrography for cases of late Perthes disease where valgus osteotomy may be appropriate or for those unusual or questionable cases where clinical findings are inconclusive and therefore the type or appropriateness of treatment is uncertain. PMID- 17414782 TI - Valgus slipped capital femoral epiphysis without posterior displacement: two case reports. AB - Valgus slipped capital femoral epiphysis is a rare entity; approximately, 26 cases in 31 hips have been reported in the orthopedic literature since 1926. It has been argued that epiphyseal displacement occurs only in a posterior or posteromedial direction, and that the radiographic appearance of true valgus or lateral slippage is an optical illusion owing to the rotation of the femur. We present the cases of two sisters with bilateral lateral slipped capital femoral epiphysis without a posterior component of displacement. In these cases, the radiographs suggested that the direction of displacement was in the lateral plane and was confirmed with computerized tomography and three-dimensional reconstructions. Analysis demonstrated the absence of posterior slippage of the proximal femoral epiphysis. The slippage was treated with percutaneous screw placement. In cases in which the direction of the slip may be in question, advanced imaging techniques may be useful for precise determination of the screw placement. This may assist in the preoperative planning, and reduce the risk of inappropriate screw placement. PMID- 17414783 TI - Long-term follow-up of the surgically corrected clubfoot. AB - We describe a 17-year follow-up of 44 surgically corrected clubfeet. They were classified preoperatively as mild, moderate or severe and grouped into those operated on before or after 3 months of age. Moderate and severe deformities underwent a posteromedial release after a trial of conservative management. The mean Ponseti score for both groups was 83.9 (good). Severely deformed feet achieved a more favourable result when operated upon before 3 months than those operated later. We conclude that early application of surgery yields better results with severely deformed feet and produces a good functional outcome in the majority of feet. PMID- 17414784 TI - Calcaneal derotation osteotomy for clubfoot revision surgery. AB - To describe a derotation calcaneal osteotomy technique and assess its efficacy in the correction of relapsed clubfoot. Twenty-one osteotomies were performed in 20 children with recurrent clubfoot. Nineteen children had been previously treated operatively. The derotation osteotomy was the first procedure performed in one case. The procedure combined medial and plantar releases, followed by a curvilinear osteotomy of the calcaneus. Patients were evaluated clinically and with standing dorsoplantar and lateral radiographs preoperatively, postoperatively and at follow-up. The talocalcaneal angle was considered as the most important criteria to evaluate the deformity correction. The clubfoot was idiopathic in 16 cases and neurological in five cases. Mean age at surgery was 7 years old (range 3.4-12 years). Total number of procedures per foot averaged 2.4 (range 4-1). The mean postoperative follow-up period was 2.8 years (range 2-6 years). The talocalcaneal angle increased significantly after the procedure (P<0.001), and no significant loss of correction was seen at latest follow-up (P=0.17). Two scarring complications occurred. Only one foot underwent further surgery after the calcaneal osteotomy for residual forefoot adduction, associated to a cavus and severe fibrosis. The calcaneal curvilinear osteotomy, in which the calcaneoforefoot unit derotation is performed around the talus but within the calcaneus, is a safe and efficient technique that can be proposed for clubfoot revision surgery. PMID- 17414785 TI - Acute correction of severe neglected club feet using a circumferential incision. AB - Severe neglected club feet in older children are a problem frequently seen in less privileged countries. Gradual correction using external fixation is a reliable method but needs sufficient infrastructure and professional supervision for follow-up. Acute corrections may be justified if such infrastructure is not available and a single stage solution is needed. A circumferential fasciocutaneous flap has been developed to allow skin coverage for acute corrections, which has been used besides a modification of other flaps used in correction of severe club feet. PMID- 17414786 TI - Polydactyly of the foot. AB - Fifty-six patients with 72 duplicated toes were analysed. Postaxial duplication accounted for 79%, and the most common anatomical pattern was duplication of the proximal phalanx with a wide metatarsal head. Forty-two patients with 55 duplications were clinically and radiographically evaluated at long-term follow up (mean 22.5 years). Results were satisfactory in 91% of the patients. Poor results were often associated with preaxial polydactyly because of persistent hallux varus. Surgical treatment is usually straightforward but must be individualized, and some anatomical and surgical details should be considered to obtain a better result. PMID- 17414787 TI - Prevention of ulnar nerve injury during fixation of supracondylar fractures in children by 'flexion-extension cross-pinning' technique. AB - No consensus exists concerning the best pin configuration for displaced supracondylar fractures of the humerus in children. Although cross-pinning is the most stable biomechanically, this configuration may cause iatrogenic ulnar nerve palsy. For the last 7 years, we have been using a three-pin fixation technique with insertion of two K wires from the lateral side (elbow in full flexion) and the third wire through the medial side (elbow in full extension). We used this technique in 67 displaced supracondylar fractures without any complications related to the ulnar nerve. The technique provides excellent stability and eliminates the risk of iatrogenic ulnar nerve palsy. PMID- 17414788 TI - Carpal tunnel syndrome in lysosomal storage disorders: simple decompression or external neurolysis? AB - Thirty-two patients (61 limbs) with lysosomal storage disorders underwent surgery for 'carpal tunnel syndrome'. Twenty-two limbs underwent a simple decompression whereas 39 limbs underwent additional neurolysis and tenosynovectomy. Data were incomplete for six patients (12 limbs). The mean age at operation for the decompression group (11 patients) was 10.5 years and for the neurolysis group (15 patients), 6.9 years. Overall, using a neurophysiological rating system, 39% of limbs demonstrated sensory improvement and 47% motor improvement with no significant difference between the groups. Only in the neurolysis group was a decline in motor conduction (2/29 limbs) or sensory conduction (four limbs) noted. Using neurophysiological criteria, no added benefit from external neurolysis was identified. PMID- 17414789 TI - Unilateral hypoplasia of the trapezius muscle in a 10-year-old boy: a case report. AB - We present a 10-year-old boy with a partial absence of or a hypoplastic right trapezius. At present, his only concern is shoulder asymmetry. No family history of significance and no history of trauma exist. His radiographs confirm changes in bony anatomy secondary to the altered balance of muscle forces on the skeleton. We have not identified any other clinical report of a partial or total absence of the trapezius although it has been defined in cadaveric cases. Similarly, some papers have described an absence of trapezius in combination with other abnormalities. In these cases, an abnormal blood supply has been described in contrast to the normal neurovascular anatomy identified in the cadaveric cases with partial absence. If this patient develops painful disability, the Eden-Lange procedure may be an appropriate treatment as for patients with spinal accessory nerve palsies. PMID- 17414790 TI - Limb lengthening by the Vilarrubias method: the Sheffield Children's Hospital experience. AB - The Vilarrubias method of limb lengthening is a percutaneous procedure with soft tissue releases. This reduces soft tissue tension and prevents joint contractures during longer lengthenings. This method has been used for 99 lower limb lengthenings. The mean length gained was 92 mm and the mean Bone Healing Index was 41.3 days/cm. There were 45 pin site problems, 33 flexion contractures, 33 angular deformities and 15 stress fractures. This complication rate compares well with other techniques. We recommend that the Vilarrubias method can be considered for lengthenings, where the aim is greater than 15% of the bone length with potential soft tissue problems. PMID- 17414791 TI - The effects of mass immunization on Haemophilus influenzae type B-related orthopaedic disease. AB - Haemophilus influenzae type B has been responsible for a significant proportion of cases of septic arthritis in children in the past. This study looks at the effects of vaccination on childhood orthopaedic infection. Data were collected from 1988 by the Public Health Laboratory Services. Data were analysed with a two sample t-test. A total of 16 of 17 cases were attributed to H. influenzae type B. Fourteen cases occurred in 5 years before mass immunization. Only two cases occurred in the 8 years following immunization. The incidence of H. influenzae septic arthritis in children has fallen significantly since the introduction of immunization (P=0.009). Vaccination has resulted in a significant fall in the incidence of H. influenzae type B septic arthritis in children. PMID- 17414792 TI - Surgical treatment of scoliosis associated with central core disease: minimizing the effects of malignant hyperthermia with provocation tests. AB - We report the case of a 7-year-old boy with thoracolumbar scoliosis and central core disease who had a history of malignant hyperthermia. He had scoliosis with Cobb's angle deteriorating to 67 degrees (thoracic) and 59 degrees (lumbar). A provocation test of general anesthesia was performed to confirm no hyperthermic reaction. Then, he underwent surgical correction by a posterior approach. The thoracic curve was reduced to 38 degrees and the lumbar curve to 42 degrees . He has been followed up for 2 years without any complications. This is the first report with a detailed description of perioperative management on surgical treatment of scoliosis associated with central core disease. PMID- 17414793 TI - Incidence, etiology, and symptomatology of upper respiratory illness in elite athletes. AB - PURPOSE: Upper respiratory illness (URI) is the most common medical condition affecting elite athletes. The aims of this study were to identify and evaluate the incidence, pathogenic etiology, and symptomatology of acute URI during a 5 month training and competition period. METHODS: Thirty-two elite and 31 recreationally competitive triathletes and cyclists, and 20 sedentary controls (age range 18.0-34.1 yr) participated in a prospective surveillance study. Nasopharyngeal and throat swabs were collected from subjects presenting with two or more defined upper respiratory symptoms. Swabs were analyzed using microscopy, culture, and PCR testing for typical and atypical respiratory pathogens. The Wisconsin Upper Respiratory Symptom Survey (WURSS-44) was used to assess symptomatology and functional impairment. RESULTS: Thirty-seven URI episodes were reported in 28 subjects. Incidence rate ratios for illness were higher in both the control subjects (1.93, 95% CI: 0.72-5.18) and elite athletes (4.50, 1.91 10.59) than in the recreationally competitive athletes. Infectious agents were identified in only 11 (two control, three recreationally competitive, and six elite) out of 37 illness episodes. Rhinovirus was the most common respiratory pathogen isolated. Symptom and functional impairment severity scores were higher in subjects with an infectious pathogen episode, particularly on illness days 3 4. CONCLUSION: The results confirm a higher rate of URI among elite athletes than recreationally competitive athletes during this training and competition season. However, because pathogens were isolated in fewer than 30% of URI cases, further study is required to uncover the causes of unidentified but symptomatic URI in athletes. Despite the common perception that all URI are infections, physicians should consider both infectious and noninfectious causes when athletes present with symptoms. PMID- 17414794 TI - Exercise training-induced changes in coagulation factors in older adults. AB - The coagulation cascade plays a critical role in the development of cardiovascular disease (CVD). Elevated plasma prothrombin fragment 1 + 2 (F1 + 2) and factor VIII antigen (FVIII:Ag) levels have been associated with a hypercoagulable state, enhancing the risk for vascular thrombotic events. Aerobic training is known to reduce CVD risk, and an improved coagulation profile may contribute to this reduction. PURPOSE: To analyze the effect of 6 months of standardized aerobic exercise training on resting F1 + 2 and FVIII:Ag levels in men and postmenopausal women aged 50-75 while accounting for several possibly confounding factors. MATERIALS AND METHODS: Sedentary men (N=16) and women (N=31) underwent supervised aerobic training 3 d x wk(-1) for 6 months while maintaining the American Heart Association step 1 diet. Baseline and final testing included measurement of F1 + 2, FVIII:Ag, plasma lipoprotein-lipid levels, body composition, and VO2max. RESULTS: When adjusted for baseline values and changes in diastolic blood pressure with training, F1 + 2 was found to decrease significantly with exercise training from 1.493 +/- 0.058 to 1.422 +/- 0.059 nM (P=0.014). FVIII:Ag levels were found to increase significantly with training when adjusted for baseline values, from 152.5 +/- 6.7% of standard at baseline to 156.0 +/- 6.1% of standard at final testing (P=0.005). Training-induced changes in coagulation markers were independent of changes in blood lipids, aerobic capacity, and body composition. CONCLUSIONS: : These results indicate that endurance training has a significant impact on the coagulation cascade, reducing coagulation activity in the common pathway and thrombin formation at rest while increasing the activation potential of the intrinsic pathway. PMID- 17414795 TI - Effect of free-living daily physical activity on salivary secretory IgA in elderly. AB - PURPOSE: The aim of this work was to assess the relationship in elderly subjects between free-living daily physical activity and mucosal immunity, especially salivary secretory immunoglobulin A (SIgA). METHODS: Elderly volunteers (114 men and 170 women) aged 71.3 +/- 0.3 yr (range: 65-86 yr) participated in this study. Resting saliva samples were collected in the morning. Saliva samples stimulated by chewing a sterile cotton ball at a frequency of 60/60 s were collected. The SIgA concentration was measured using enzyme-linked immunosorbent assay (ELISA), and the SIgA secretion rate was calculated. Free-living step count (steps per day), energy expenditure (kJ x kg(-1) x d(-1)), and activity durations (min x d( 1)) at specific intensity levels (inactive, light, moderate, and vigorous) were evaluated using an electric pedometer. The data obtained were stratified by pedometer-determined steps per day using quartiles (Q1-Q4) for distribution. RESULTS: Elderly in quartiles showed step counts of 2962 +/- 94, 5118 +/- 62, 6832 +/- 59, and 9951 +/- 264 steps per day. Significant differences were found in the mean step count (P<0.0001), energy expenditure (P<0.0001), and activity duration (P<0.0001) with increasing pedometer-determined activity quartiles. Both SIgA concentration and SIgA-secretion rate were significantly higher for Q3 than for Q1 (P<0.05). Meanwhile, saliva flow rates showed no significant differences across quartiles. CONCLUSION: These results suggest that a free-living daily physical activity level of approximately 7000 steps per day might be regarded as a moderate daily physical activity target for elderly people to improve mucosal immune function. PMID- 17414796 TI - Depression and pain in retired professional football players. AB - PURPOSE: To assess the prevalence of depressive symptoms and difficulty with pain in retired professional football players, difficulties with the transition from active athletic competition to retirement, perceptions of barriers to receiving assistance for those difficulties, and recommended programs to provide such assistance. METHODS: Survey sent to 3377 retired members of the National Football League Players Association (NFLPA), with usable responses received from 1617 members (functional response rate, 48.6%). RESULTS: Respondents were categorized as experiencing no to mild depression (N=1366; 84.5%) or moderate to severe depression (N=237; 14.7%). Respondents were also categorized according to whether they reported difficulty with pain as not or somewhat common (N=837; 51.8%) versus quite or very common (N=769; 47.6%). Respondents most frequently reported trouble sleeping, financial difficulties, marital or relationship problems, and problems with fitness, exercise, and aging, all of which were strongly correlated with the presence of moderate to severe depression and with quite or very common difficulty with pain. The same difficulties were even more commonly experienced by respondents who reported both moderate to severe depression and quite or very common difficulty with pain, compared with those who reported low scores in both domains. CONCLUSION: Retired professional football players experience levels of depressive symptoms similar to those of the general population, but the impact of these symptoms is compounded by high levels of difficulty with pain. The combination of depression and pain is strongly predictive of significant difficulties with sleep, social relationships, financial difficulties, and problems with exercise and fitness. A hypothesis explaining this association is that significant musculoskeletal disability and chronic pain interferes with physical activity and fitness during retirement and increases the risk of depression. PMID- 17414797 TI - Physical activity, obesity status, and glycemic control: The ATTICA study. AB - PURPOSE: We aimed to evaluate the relationship of physical activity and obesity with glycemic control and insulin resistance. METHODS: A randomized, population based, cross-sectional health and nutrition survey was conducted in the province of Athens, Greece. Subjects included 1514 men and 1528 women without evidence of cardiovascular or other chronic disease. Participants were classified as inactive, minimally active, or health-enhancing physical activity (HEPA) active based on the International Physical Activity Questionnaire. Insulin sensitivity was assessed by the homeostatic model (HOMA), and overweight or obesity was assessed according to BMI (BMI >or= 25). Related social, biological, and lifestyle factors were also recorded and used as potential confounders. RESULTS: Five hundred sixty-five (37.3%) men and 493 (32.3%) women were classified as physically active. From the 1058 (34.8%) subjects who were classified as active, 306 (10.1%) met the criteria for HEPA active, and the rest were minimally active. HEPA active and minimally active subjects smoked less and had lower BMI, waist, and waist-to-hip ratio. Lean and overweight or obese subjects with sedentary lifestyle had greater levels of glucose, insulin, and insulin resistance [corrected] However, overweight or obese volunteers with physical activity levels classified as HEPA had similar levels of glucose and insulin sensitivity, with lower insulin than lean inactive individuals. Linear regression analysis between HOMA and physical activity, taking into consideration several social and biological factors, showed that physical activity (MET x min x wk(-1)), age, BMI, and total energy intake are important predictors of HOMA, whereas other factors such as waist circumference did not reach statistical significance. CONCLUSION: Our data show that physical activity is a significant factor on insulin sensitivity, whereas increased physical activity may ameliorate the well-known effects of obesity on insulin sensitivity. PMID- 17414798 TI - Reliability and validity of questions on the youth media campaign longitudinal survey. AB - PURPOSE: The study evaluated the reliability and validity of the physical activity questions in the Youth Media Campaign Longitudinal Survey (YMCLS), a nationally representative survey of 9- to 13-yr-old youth. METHODS: The participants were 192 youth aged 9-13 yr (93 males and 99 females) in grades 4-8 from eight schools in a large, ethnically diverse school district. Participants completed two YMCLS phone interviews, which were administered 1 wk apart by trained interviewers. Reliability was examined by comparing data from two administrations of the survey. Validity was examined by comparing YMCLS measures from the second administration with temporally matched measures from an accelerometer and a detailed activity log. RESULTS: Reliability coefficients were high for estimates of organized activity (intraclass correlation coefficient (ICC)=0.78) and moderate for estimates of free-time activity (ICC=0.60) and total weekly activity (ICC=0.60). Estimates of total weekly activity sessions were significantly correlated with the accelerometer (r=0.24) and the activity log (r=0.46). Estimates of activity time and activity sessions on the previous day were also significantly correlated with the accelerometer (r=0.53 and 0.37, respectively) and the activity log (r=0.37 and 0.47, respectively). Correlations between the YMCLS and the activity log were higher for organized activity (r=0.72) than for free-time activity (r=0.46). Reliability and validity coefficients were similar for boys and girls, but older youth (11-13 yr) had higher coefficients than younger students (9-10 yr). CONCLUSION: The YMCLS has acceptable reliability and validity for estimating habitual physical activity in youth aged 9-13 yr. PMID- 17414799 TI - Intraindividual variation of objectively measured physical activity in children. AB - PURPOSE: This study examined the seasonal and intraindividual variation in objectively measured physical activity in 11- to 12-yr-olds. METHODS: Children were asked to wear a uniaxial accelerometer for 7 d four times throughout the course of about a year. A random-intercepts model was used to separate the inter- and intraindividual components of physical activity. Gender, age, body mass index (BMI), height, and month of measurement were fitted to the model as potential confounders. RESULTS: A total of 315 children had valid data for at least two measurement occasions, and 244 had data for all four measurement occasions. The unadjusted intraclass correlation coefficient (ICC) for total activity (counts per minute) was 0.54; 0.49 after adjusting for gender, age, and BMI; and 0.53 after adjusting for gender, age, BMI, and month. Further adjustment for pubertal status at baseline had no effect on the ICC. Restricting the analysis to only those with data for all four measurement occasions (N=244), or to measurements taken on school days only, had no effect on the ICC. The fully adjusted ICC was 0.51 for weekdays only and 0.39 for weekend days only. For minutes of moderate to vigorous physical activity, minutes of vigorous activity, minutes of sedentary behavior, and number of 30-min blocks of sedentary behavior, the fully adjusted ICC were 0.45, 0.37, 0.59, and 0.39, respectively. The analysis was repeated for boys and girls separately, but the differences in ICC were small. CONCLUSION: There was substantial intraindividual variation in the objectively measured physical activity of these children. Studies using single a measurement occasion where physical activity is the exposure should take this into account to adjust for regression dilution. PMID- 17414800 TI - Comparison of Kenz Lifecorder EX and ActiGraph accelerometers in 10-yr-old children. AB - A new accelerometer, the Kenz Lifecorder EX (LC; Suzuken Co. Ltd, Nagoya, Japan), offers promise as a feasible monitor alternative to the commonly used Actigraph (AG: Actigraph LLC, Fort Walton Beach, FL). PURPOSE: This study compared the LC and AG accelerometers and the Yamax SW-200 pedometer (DW) under free-living conditions with regard to children's steps taken and time in light-intensity physical activity (PA) and moderate to vigorous PA (MVPA). METHODS: Participants (N=31, age=10.2 +/- 0.4 yr) wore LC, AG, and DW monitors from arrival at school (7:45 a.m.) until they went to bed. Time in light and MVPA intensities were calculated using two separate intensity classifications for the LC (LC_4 and LC_5) and four classifications for the AG (AG_Treuth, AG_Puyau, AG_Trost, and AG_Freedson). Both accelerometers provided steps as outputs. DW steps were self recorded. Repeated-measures ANOVA was used to assess overlapping monitor outputs. RESULTS: There was no difference between DW and LC steps (Delta=200 steps), but a nonsignificant trend was observed in the pairwise comparison between DW and AG steps (Delta=1001 steps, P=0.058). AG detected significantly greater steps than the LC (Delta=801 steps, P=0.001). Estimates of light-intensity activity minutes ranged from a low of 75.6 +/- 18.4 min (LC_4) to a high of 309 +/- 69.2 min (AG_Treuth). Estimates of MVPA minutes ranged from a low of 25.9 +/- 9.4 min (LC_5) to a high of 112.2 +/- 34.5 min (AG_Freedson). No significant differences in MVPA were seen between LC_5 and AG_Treuth (Delta=4.9 min) or AG_Puyau (Delta=1.7 min). CONCLUSION: The LC detected a comparable number of steps as the DW but significantly fewer steps than the AG in children. Current results indicate that the LC_5 and either AG_Treuth or AG_Puyau intensity derivations provide similar mean estimates of time in MVPA during-free living activity in 10 yr-old children. PMID- 17414801 TI - Oat beta-glucan effects on neutrophil respiratory burst activity following exercise. AB - Fatiguing exercise has been associated with a decrease in certain functions of neutrophils, whereas moderate exercise has generally been associated with an increase. Consumption of oat beta-glucan (ObetaG), a soluble fiber and mild immune system enhancer, may offset the immunosuppression associated with intense training and perhaps further enhance the benefits of moderate exercise. PURPOSE: To test the effects of ObetaG consumption on neutrophil function and number after both moderate and fatiguing exercise. METHODS: Male mice were assigned to one of six treatment groups. Fatiguing exercise mice (Ftg-ObetaG and Ftg-H2O) ran to volitional fatigue on a treadmill for three consecutive days, and moderate exercise mice (Mod-ObetaG and Mod-H2O) ran for six consecutive days for 1 h. Control mice (Con-ObetaG and Con-H2O) were exposed to the treadmill environment but did not run. ObetaG was consumed in the drinking water (approximately 0.6 mL x d(-1)) for 10 consecutive days. After rest or exercise on the last day of training, mice were given a 1-mL i.p. injection of thioglycollate. Mice were sacrificed 3 h later; neutrophils were harvested from the peritoneal cavity and counted, and their respiratory burst activity was measured using flow cytometry. RESULTS: Both moderate exercise and ObetaG increased neutrophil burst activity, whereas fatiguing exercise had no effect. Neutrophil number was increased by fatiguing exercise and ObetaG, but not moderate exercise. There were no additive effects of exercise and ObetaG on either of these variables. CONCLUSION: These data suggest that although not additive in their effects, both ObetaG and exercise can alter overall neutrophil respiratory burst activity (number and/or function), but only ObetaG increased both number and function, which may have important ramifications for defense against infection. PMID- 17414802 TI - Antioxidant supplementation and immunoendocrine responses to prolonged exercise. AB - PURPOSE: Antioxidant supplementation may modulate systemic cortisol and interleukin-6 (IL-6) responses to prolonged exercise, but it is unclear whether such effects are also associated with a reduction in the magnitude of immunodepression. The purpose of the present study was to examine the effects of daily vitamin C (L-ascorbic acid, 1000 mg x d(-1)) and vitamin E (RRR-alpha tocopherol, 400 IU x d(-1)) supplementation on immunoendocrine responses to prolonged exercise. METHODS: Twenty healthy, recreationally active males cycled for 2.5 h at approximately 60% of maximal oxygen uptake after 4 wk of placebo (PLA, N=10) or antioxidant (AO, N=10) supplementation. RESULTS: A significant group x time interaction was observed for plasma cortisol concentration (P=0.008), and the postexercise increase was greater (P<0.05) in the PLA compared with AO group (approximately 170% compared with an approximately 120% increase above baseline). Plasma IL-6 concentration was significantly increased after exercise to a similar extent in both groups. Plasma free F2-isoprostane concentration was significantly increased after exercise and was unaffected by AO supplementation, whereas plasma TBARS was unaffected by exercise in the PLA group but was lower after exercise in the AO group than in the PLA group. Circulating neutrophil count was significantly increased after exercise, and in vitro bacteria-stimulated elastase release per neutrophil was significantly decreased to a similar extent in both groups. CONCLUSIONS: These results suggest that 4 wk of AO supplementation may blunt the cortisol response to a single 2.5-h bout of prolonged exercise independently of changes in oxidative stress or plasma IL-6 concentration, but it is not effective at modulating the exercise-induced neutrophilia or depression of neutrophil function. PMID- 17414803 TI - Effects of eccentric exercise on NF-kappaB activation in blood mononuclear cells. AB - PURPOSE: Changes in nuclear factor kappaB (NF-kappaB) activation induced in peripheral blood mononuclear cells (PBMC) by acute eccentric exercise and by submaximal eccentric training were investigated. METHODS: Eleven subjects carried out two bouts of eccentric exercise separated by 6 wk of training. RESULTS: Soreness, vertical jump height, and plasma creatine kinase were significantly modified after the first bout. NF-kappaB activation, p50 and p65, phospho IkappaBalpha and phospho-IKK protein level, and Mn-SOD expression increased in PBMC, whereas IkappaBalpha protein level was significantly reduced. Changes were significantly attenuated after the second exercise bout. An additional group of nine subjects carried out the two bouts of exercise without training. Effects on NF-kappaB activation were similar after the second bout compared with the first, despite a reduction in markers of muscle injury (repeated bout effect). CONCLUSION: Training significantly attenuates the NF-kappaB-dependent pathway changes induced in PBMC by eccentric exercise, with no contribution from the repeated bout effect. PMID- 17414804 TI - Aerobic high-intensity intervals improve VO2max more than moderate training. AB - PURPOSE: The present study compared the effects of aerobic endurance training at different intensities and with different methods matched for total work and frequency. Responses in maximal oxygen uptake (VO2max), stroke volume of the heart (SV), blood volume, lactate threshold (LT), and running economy (CR) were examined. METHODS: Forty healthy, nonsmoking, moderately trained male subjects were randomly assigned to one of four groups:1) long slow distance (70% maximal heart rate; HRmax); 2)lactate threshold (85% HRmax); 3) 15/15 interval running (15 s of running at 90-95% HRmax followed by 15 s of active resting at 70% HRmax); and 4) 4 x 4 min of interval running (4 min of running at 90-95% HRmax followed by 3 min of active resting at 70%HRmax). All four training protocols resulted in similar total oxygen consumption and were performed 3 d.wk for 8 wk. RESULTS: High-intensity aerobic interval training resulted in significantly increased VO2max compared with long slow distance and lactate-threshold training intensities (P<0.01). The percentage increases for the 15/15 and 4 x 4 min groups were 5.5 and 7.2%, respectively, reflecting increases in V O2max from 60.5 to 64.4 mL x kg(-1) x min(-1) and 55.5 to 60.4 mL x kg(-1) x min(-1). SV increased significantly by approximately 10% after interval training (P<0.05). CONCLUSIONS: : High-aerobic intensity endurance interval training is significantly more effective than performing the same total work at either lactate threshold or at 70% HRmax, in improving VO2max. The changes in VO2max correspond with changes in SV, indicating a close link between the two. PMID- 17414805 TI - Exercise-induced decrease in insular cortex rCBF during postexercise hypotension. AB - The insular cortex (IC), a region of the brain involved in blood pressure (BP) modulation, shows decreases in regional cerebral blood flow (rCBF) during postexercise hypotension (PEH). PURPOSE: To determine whether changes in IC neural activity were caused by prior exercise or by changes in BP, this investigation compared patterns of rCBF during periods of hypotension, which was induced by prior exercise (i.e., PEH) and sodium nitroprusside (SNP) infusion and a cold pressor (CP), to restore BP. METHODS: Ten subjects were studied on three different days with randomly assigned conditions: i) resting baseline; ii) PEH; and iii) SNP-induced hypotension (matched to the PEH BP decrease). Data were collected for heart rate (HR) and mean BP, and rCBF was assessed using single photon emission computed tomography (SPECT) as an index of brain activation. RESULTS: Using ANOVA across conditions, there were differences (P<0.05; mean +/- SD) from baseline during PEH for HR (+12 +/- 3 bpm) and mean BP (-8 +/- 2 mm Hg) and during SNP-induced hypotension (HR = +15 +/- 4 bpm; MBP = -9 +/- 2 mm Hg), with no differences between PEH and SNP. After exercise, there were decreases (P<0.05) in the leg sensorimotor area, anterior cingulate, and the right and left inferior thalamus, right inferior insula, and left anterior insular regions. During SNP-induced hypotension, there were significant increases in the right and left inferior thalamus and the right and left inferior anterior IC. CP during PEH increased BP and IC activity. CONCLUSIONS: Data show that reductions in IC neural activity are not caused by acute BP decreases. Findings suggest that exercise can lead to a temporary decrease in IC neural activity, which may be a significant neural factor contributing to PEH. PMID- 17414806 TI - Why does power output decrease at high pedaling rates during sprint cycling? AB - PURPOSE: The objective of this study was to partly explain, from electromyographical (EMG) activity, the decrease in power output beyond optimal pedaling rate (PRopt) during sprint cycling. METHODS: Eleven cyclists performed four 8-s nonisokinetic sprints on a cycle ergometer against four randomized friction loads (0.5, twice 0.75, and 0.9 N x kg(-1) of body mass). Power output and EMG activity of both right and left gluteus maximus, rectus femoris, biceps femoris, and vastus lateralis were measured continuously. Individual crank cycles were analyzed. Crank angles corresponding to the beginning and the peak of each downstroke and EMG burst onset and offset crank angles were computed. Moreover, crank angles corresponding to the beginning and the end of muscle force response were determined assuming a 100-ms lag time between the EMG activity and the relevant force response (or electromechanical delay). RESULTS: Muscle coordination (EMG onset and offset) was altered at high pedaling rates. Thus, crank angles corresponding to muscle force response increased significantly with pedaling rate. Consequently, at pedaling rates higher than the optimal pedaling rate, force production of lower-limb extensor muscles was shifted later in the crank cycle. Mechanical data confirmed that downstrokes occurred later in the crank cycle when pedaling rate increased. Hence, force was produced on the pedals during less effective crank cycle sectors of the downstroke and during the beginning of the upstroke. CONCLUSION: During nonisokinetic sprint cycling, the decrease in power output when pedaling rates increased beyond PRopt may be partly explained by suboptimal muscle coordination. PMID- 17414807 TI - High REE in Sumo wrestlers attributed to large organ-tissue mass. AB - PURPOSE: It is unknown whether high resting energy expenditure (REE) in athletes is attributable to changes in organ-tissue mass and/or metabolic rate. The purpose of this study was to examine the contribution of organ-tissue mass of fat free mass (FFM) components to REE for Sumo wrestlers who have large FFM and REE. We investigated the relationship between the REE measured by indirect calorimetry and the REE calculated from organ-tissue mass using a previously published approach. METHODS: Ten Sumo wrestlers and 11 male untrained college students (controls) were recruited to participate in this study. FFM was estimated by two component densitometry. Contiguous magnetic resonance imaging (MRI) images with a 1-cm slice thickness were obtained from the top of head to the ankle joints, and the cross-sectional area and volume were determined for skeletal muscle (SM), liver, kidney, and brain. The volume of adipose tissue, heart, and residual was calculated from each equation. The volume units were converted into mass by an assumed constant density. The measured REE was determined by indirect calorimetry. The calculated REE was estimated as the sum of individual organ tissue masses (seven body compartments) multiplied by their metabolic rate constants. RESULTS: The measured REE for Sumo wrestlers (2286 kcal x d(-1)) was higher (P<0.01) than for controls (1545 kcal x d(-1)). Sumo wrestlers had a greater amount of FFM and FFM components (e.g., SM, liver, and kidney), except for brain. The ratio of measured REE to FFM and the measured REE adjusted by FFM were similar between the two groups. The measured REE values for Sumo wrestlers were not significantly different from the calculated REE values. CONCLUSIONS: The high REE for Sumo wrestlers can be attributed not to an elevation of the organ tissue metabolic rate, but to a larger absolute amount of low and high metabolically active tissue including SM, liver, and kidney. PMID- 17414808 TI - Gastrocnemius medialis and vastus lateralis oxygenation during whole-body vibration exercise. AB - PURPOSE: The aim of this study was to investigate the effects of different whole body vibration (WBV) frequencies on oxygenation of vastus lateralis (VL) and gastrocnemius medialis (GM) muscles during static squatting in sedentary and physically active healthy males. METHODS: Twenty volunteers (age: 24.6 +/- 2.9 yr; body mass: 80.6 +/- 11.8 kg; height: 178.1 +/- 7.6 cm) participated in this study. Ten subjects were sedentary individuals and 10 were athletes practicing different sports. All subjects completed four trials (control, and 30-, 40-, and 50-Hz WBV) in a randomized controlled crossover design. The trials consisted of static squatting on a vibrating platform for a total duration of 110 s. Muscle oxygenation status was recorded with near-infrared spectroscopy. RESULTS: The data analysis revealed no significant treatment-by-time interactions in tissue oxygenation index (TOI) or Delta total hemoglobin volume (tHb) in VL and GM muscles. A significant main effect of time in TOI of both VL and GM muscles was identified (P<0.001). VL TOI significantly decreased by 2.8% at 90 s in the control condition and by 3.3% at 110 s in the 30-Hz condition; VL TOI significantly increased by 2.1 and 3.0% at 30 s in the 40- and 50-Hz conditions, respectively. GM TOI significantly decreased by 3.2% at 60 s, by 4.1% at 90 s, and by 4.3% at 110 s in the control condition, and by 5.5% at 110 s in the 30-Hz condition. CONCLUSION: This study showed that WBV exercise with frequencies of 30, 40, and 50 Hz and small amplitudes does not affect muscle oxygenation of VL and GM muscles to a higher degree than a nonvibration condition. PMID- 17414809 TI - BMI and health-related physical fitness in Taiwanese youth 9-18 years. AB - PURPOSE: The purpose of the study was to evaluate the cross-sectional relationship between BMI and a physical fitness index (PFI) based on four indicators of fitness in a national sample of Taiwanese youth. METHODS: Height, weight, and four measures of physical fitness (sit-ups completed in 60 s, standing long jump, sit and reach, and 800- or 1600-m run/walk) were measured in a national sample of 102,765 Taiwanese youth 9-18 yr of age: 50,940 girls and 51,825 boys. BMI was calculated for each subject. Within each sex-specific half year age group, students were classified into five BMI categories based on national percentiles: very low, BMI < 5th percentile; low, BMI >or= 5th but < 15th percentiles; normal, BMI >or= 15th but < 85th percentiles; high, BMI >or= 85th but < 95th percentiles; and very high, BMI >or= 95th percentiles. Z-scores based on sex- and age-specific means and standard deviations were calculated, and the sum of z-scores for the four fitness tests was used as a PFI. Differences in PFI between BMI categories within each sex-specific half-year age group were compared with ANOVA with Bonferroni adjustments. Sex-specific regressions of PFI on BMI, using a nonlinear quadratic model, were done in four broader age categories. RESULTS: Relationships between BMI and PFI are nonlinear and vary with age from late childhood through adolescence. With increasing age during adolescence, the relationship becomes parabolic, and the peaks of the parabola are sharper in adolescent boys than girls. CONCLUSION: PFI declines in a curvilinear manner with increasing BMI among youth 9-18 yr of age, but the slope of the relationship varies with age. PMID- 17414810 TI - The effect of extrinsic motivation on cycle time trial performance. AB - PURPOSE: Athletes occasionally follow pacing patterns that seem unreasonably aggressive compared with those of prerace performances, potentially because of the motivation provided by competition. This study evaluated the effect of extrinsic motivation on cyclists' time trial performance. METHODS: Well-trained recreational cyclists (N=7) completed four 1500-m laboratory time trials including a practice trial, two self-paced trials, and a trial where a monetary reward was offered. Time, total power output, power output attributable to aerobic and anaerobic metabolic sources, VO2, and HR were measured. RESULTS: The time required for the second, third, and last (extrinsically motivated) time trials was 133.1 +/- 2.1, 134.1 +/- 3.4, and 133.6 +/- 3.0 s, respectively, and was not different (P>0.05). There were no differences for total (396 +/- 19, 397 +/- 23, and 401 +/- 17 W), aerobic (253 +/- 12, 254 +/- 10, and 246 +/- 13 W), and anaerobic (143 +/- 14, 143 +/- 21, and 155 +/- 11 W) power output. The highest VO2 was not different over consecutive time trials (3.76 +/- 0.19, 3.73 +/- 0.16, and 3.71 +/- 0.22 L x min(-1)). When ranked by performance, without reference to the extrinsic motivation (131.9 +/- 2.4, 133.4 +/- 2.4, and 135.4 +/ 2.5 s), there was a significant difference for the first 100 m and from 100 to 300 m in power output, with a larger total power (560 +/- 102, 491 +/- 82, and 493 +/- 93; and 571 +/- 94, 513 +/- 41, and 484 +/- 88 W) and power attributable to anaerobic sources (446 +/- 100, 384 +/- 80, and 324 +/- 43; and 381 +/- 87, 383 +/- 90, and 289 +/- 91 W) for the fastest trial. CONCLUSION: Extrinsic motivation did not change the time trial performance, suggesting that 1500-m performance is extremely stable and not readily changeable with simple external motivation. The results suggest that spontaneous improvement in performance for time trials of this duration is attributable to greater early power output, which is primarily attributable to anaerobic metabolic sources. PMID- 17414811 TI - Influence of speed and step frequency during walking and running on motion sensor output. AB - PURPOSE: Studies have reported strong linear relationships between accelerometer output and walking/running speeds up to 10 km x h(-1). However, ActiGraph uniaxial accelerometer counts plateau at higher speeds. The aim of this study was to determine the relationships of triaxial accelerometry, uniaxial accelerometry, and pedometry with speed and step frequency (SF) across a range of walking and running speeds. METHODS: Nine male runners wore two ActiGraph uniaxial accelerometers, two RT3 triaxial accelerometers (all set at a 1-s epoch), and two Yamax pedometers. Each participant walked for 60 s at 4 and 6 km x h(-1), ran for 60 s at 10, 12, 14, 16, and 18 km x h(-1), and ran for 30 s at 20, 22, 24, and 26 km x h(-1). Step frequency was recorded by a visual count. RESULTS: ActiGraph counts peaked at 10 km x h(-10 (2.5-3.0 Hz SF) and declined thereafter (r=0.02, P>0.05). After correction for frequency-dependent filtering, output plateaued at 10 km x h(-1) but did not decline (r=0.77, P<0.05). Similarly, RT3 vertical counts plateaued at speeds > 10 km x h(-1) (r=0.86, P<0.01). RT3 vector magnitude and anteroposterior and mediolateral counts maintained a linear relationship with speed (r>0.96, P<0.001). Step frequency assessed by pedometry compared well with actual step frequency up to 20 km x h(-1) (approximately 3.5 Hz) but then underestimated actual steps (Yamax r=0.97; ActiGraph pedometer r=0.88, both P<0.001). CONCLUSION: Increasing underestimation of activity by the ActiGraph as speed increases is related to frequency-dependent filtering and assessment of acceleration in the vertical plane only. RT3 vector magnitude was strongly related to speed, reflecting the predominance of horizontal acceleration at higher speeds. These results indicate that high-intensity activity is underestimated by the ActiGraph, even after correction for frequency-dependent filtering, but not by the RT3. Pedometer output is highly correlated with step frequency. PMID- 17414812 TI - The effect of acute exercise on serum brain-derived neurotrophic factor levels and cognitive function. AB - Brain-derived neurotrophic factor (BDNF) is one of a family of neurotrophic factors that participates in neuronal transmission, modulation and plasticity. Previous studies using animals have demonstrated that acute and chronic exercise leads to increases in BDNF in various brain regions. PURPOSE: To determine the effects of acute exercise on serum BDNF levels in humans, and to determine the relationship between exercise intensity and BDNF responses. Additionally, the relationship between changes in BDNF and cognitive function was examined. METHODS: Fifteen subjects (25.4 +/- 1.01 yr; 11 male, 4 female) performed a graded exercise test (GXT) for the determination of VO2max and ventilatory threshold (VTh) on a cycle ergometer. On separate days, two subsequent 30-min endurance rides were performed at 20% below the VTh (VTh - 20) and at 10% above the VTh (VTh + 10). Serum BDNF and cognitive function were determined before and after the GXT and endurance rides with an enzyme-linked immunosorbent assay (ELISA) and the Stroop tests, respectively. RESULTS: The mean VO2max was 2805.8 +/- 164.3 mL x min(-1) (104.2 +/- 7.0% pred). BDNF values (pg x mL(-1)) increased from baseline (P<0.05) after exercise at the VTh + 10 (13%) and the GXT (30%). There was no significant change in BDNF from baseline after the VTh - 20. Changes in BDNF did not correlate with VO2max during the GXT, but they did correlate with changes in lactate (r=0.57; P<0.05). Cognitive function scores improved after all exercise conditions, but they did not correlate with BDNF changes. CONCLUSION: BDNF levels in humans are significantly elevated in response to exercise, and the magnitude of increase is exercise intensity dependent. Given that BDNF can transit the blood-brain barrier in both directions, the intensity-dependent findings may aid in designing exercise prescriptions for maintaining or improving neurological health. PMID- 17414813 TI - Heart rate variability as a predictor of negative mood symptoms induced by exercise withdrawal. AB - INTRODUCTION/PURPOSE: Negative mood symptoms occur frequently in sedentary populations, but individual vulnerability factors for developing these complaints have not been systematically evaluated. This investigation examined whether the autonomic nervous system (ANS) serves a role in the development of negative mood after controlled exercise withdrawal. METHODS: Forty participants (mean age of 31.3 +/- 7.5 yr, 55% women) who exercised regularly (>or= 30 min of continuous aerobic exercise at least three times a week during the past 6 months) were randomized either to withdrawal from regular aerobic exercise (N=20) or to continue regular aerobic exercise (N=20) for 2 wk. Measurements were taken before exercise withdrawal and at 2-wk follow-up. Various dimensions of negative mood were measured with the multidimensional fatigue inventory, profile of mood states, and Beck depression inventory-II. ANS activity was assessed by heart rate variability (HRV) analyses, examining low-frequency (0.04-0.15 Hz: lf) and high frequency (hf) domains (0.15-0.40 Hz). The lf/hf ratio was used as index of sympathovagal balance. Protocol adherence was documented by ambulatory activity monitoring. RESULTS: Exercise withdrawal resulted in significantly higher negative mood scores at follow-up compared with control (P<0.05). Baseline lf/hf ratios correlated with the increases in symptoms (r>0.4; P<0.05) in the exercise withdrawal group independently of gender, age, weight, baseline fitness level, and baseline symptom status. The exercise-withdrawal and control groups displayed no significant change in hf HRV, lf HRV, or lf/hf HRV during the 2 wk. CONCLUSION: Reduced parasympathetic ANS activity as measured by HRV is predictive of the development of negative mood after deprivation of usual exercise activities. No significant changes in HRV were observed during the 2-wk exercise deprivation period. These findings are relevant to the understanding of mood changes in response to short-term exercise withdrawal, such as sports injuries and recovery from medical procedures. PMID- 17414814 TI - Study findings challenge core components of a current model of exercise thermoregulation. PMID- 17414816 TI - Calculation of muscle activation using neuromuscular electrical stimulation. PMID- 17414818 TI - Reap the benefits of the student-teacher relationship. PMID- 17414819 TI - CA-MRSA education important. PMID- 17414820 TI - STI education still critical. PMID- 17414821 TI - A new breakthrough in diabetes treatment. PMID- 17414822 TI - Zonisamide as treatment for seizures. PMID- 17414823 TI - Ensuring clinical competency for NPs in acute care. PMID- 17414828 TI - Putting insomnia to rest. PMID- 17414830 TI - Challenges in chronic constipation management. PMID- 17414832 TI - Intranasal steroids and allergic rhinitis. PMID- 17414836 TI - Hepatobiliary pathology. AB - PURPOSE OF REVIEW: Recent papers on disorders of the liver and biliary tract which clarify their pathogenesis and attendant morphologic changes are highlighted. RECENT FINDINGS: The concept of 'bystander hepatitis' was cited in studies showing hepatic infiltration of CD8-positive T cells in the setting of extrahepatic infections such as influenza virus and severe acute respiratory syndrome. Diabetic liver lesions include glycogenic hepatopathy (in which poor diabetic control leads to swollen, glycogen-filled hepatocytes without fat, steatohepatitis or fibrosis) and diabetic hepatosclerosis in which there is diffuse perisinusoidal fibrosis (type IV collagen) without zonal predilection. Ground-glass hepatocellular inclusions (positive with periodic acid-Schiff stain for glycogen) were reported in three separate series of patients who were hepatitis B virus-negative, often transplant recipients, immunosuppressed and on multiple medications. A Banff consensus paper expertly compared and contrasted the histologic features which characterize the various causes of late liver allograft dysfunction. SUMMARY: Informative papers emerged this past year concerning collateral damage to the liver in extrahepatic infections, diabetic lesions and causes of liver dysfunction after transplantation, among other topics. PMID- 17414837 TI - Cholestasis and cholestatic syndromes. AB - PURPOSE OF REVIEW: This review focuses on the recent advances in cholestatic liver diseases. While there is an emphasis placed on translational and treatment focused studies, basic science studies with the greatest impact on the field are also covered. RECENT FINDINGS: Highlights include new discoveries for the role of the farsenoid X receptor and sodium-dependent taurocholate cotransporting polypeptide; new insights into the pathogenesis of progressive familial intrahepatic cholestasis type 1, biliary atresia, intrahepatic cholestasis of pregnancy, and primary biliary cirrhosis; new information for assessing prognosis in biliary atresia and primary biliary cirrhosis; and important clinical trials in intrahepatic cholestasis of pregnancy, primary biliary cirrhosis and primary sclerosing cholangitis. SUMMARY: The studies of 2006 have furthered our understanding of cholestasis and cholestatic syndromes. While we continue to add to our knowledge of pathogenesis and treatment for many of these diseases, much work remains. PMID- 17414838 TI - Inherited metabolic disease of the liver. AB - PURPOSE OF REVIEW: The past decade has seen extraordinary growth in our understanding of the pathophysiology of Wilson disease, genetic hemochromatosis and alpha-1 antitrypsin deficiency as we continue to elucidate the molecular and cellular machinery involved in their pathogenesis. The continued progress in the elaboration of the molecular biology, genetics, epidemiology, and management of these prototypical inherited metabolic diseases will be the focus of this review. RECENT FINDINGS: Wilson disease and genetic hemochromatosis involve defects in metal transport with copper and iron accumulation in hepatocytes, respectively. In alpha-1 antitrypsin deficiency, hepatocytes accumulate defective alpha-1 antitrypsin that misfolds. As a more complete picture of the molecular biology of the proteins and genes involved in transport has evolved, so has our understanding of the etiopathogenesis of these disorders and the variety of phenotypes observed. Finally, new ideas regarding the clinical management of these disorders will emerge with elucidation of the cellular basis for these diseases. SUMMARY: The recent developments detailed in this article have important implications for the future diagnosis and treatment of these diseases. Recent discoveries link molecular defects with alterations in the functional machinery of the cell, and provide new avenues for advancing the diagnosis and treatment of these disorders. PMID- 17414839 TI - Drug-induced liver disease in 2006. AB - PURPOSE OF REVIEW: To identify the key publications of 2006 dealing with drug induced liver injury. RECENT FINDINGS: When given in therapeutic doses over 14 days, acetaminophen produced significant asymptomatic elevations in alanine aminotransferase among healthy volunteers, suggesting that subclinical injury may be more common than previously thought. Acute liver failure in children was shown to differ in several important respects from that seen in adults, notably a much lower incidence of acetaminophen toxicity with nearly half of all cases being indeterminate in origin. The first cases of hepatotoxicity with telithromycin, a new class of ketolide antibiotic, were described along with reports suggesting liver injury from ezetimibe among other agents. The potential for chronic injury to develop after acute drug-induced liver injury was analyzed in a large Swedish database; 5-6% of cases were judged to become chronic, with drugs causing cholestatic injury predominating. Among well described hepatotoxins, new reports appeared with highly active antiretroviral therapy agents, herbal therapies and several antibiotics. Finally, the safe use of pravastatin and pioglitazone was demonstrated in patients with chronic liver disease in controlled clinical trials. SUMMARY: Drug-induced liver injury remains an important concern for many existing drugs as well as for agents in development. PMID- 17414840 TI - Autoimmune liver disease. AB - PURPOSE OF REVIEW: The aim of this article is to review studies that improve the diagnosis and treatment of autoimmune hepatitis and suggest new drug and molecular interventions. RECENT FINDINGS: Elderly patients have an indolent but aggressive disease that responds well to corticosteroid therapy. Variant syndromes are artificial designations that reflect uncertainties regarding the diagnostic limits of classical disease. Antibodies to cyclic citrullinated proteins and complex assays for antibodies to actin and alpha-actinin may have prognostic value. Defects in the number and function of T regulatory cells may enhance cell-mediated cytotoxicity. HLA DRB113 may be a risk factor in some North American patients, and disease outcome may be influenced by the 'dose' of alleles encoding critical residues. Screening for thiopurine methyltransferase deficiency does not predict azathioprine intolerance. Treatment until normalization of the laboratory and histological features reduces the risk of relapse by 30-50%. Adverse outcomes in pregnancy are associated with antibodies to soluble liver antigen/liver pancreas and Ro/SSA. SUMMARY: Novel serological tests may have prognostic value. Defects in the suppressor activity of regulatory T cells may promote liver injury. Genetic predispositions strongly influence disease occurrence and outcome. Laboratory and histological features should be normal prior to drug withdrawal. Azathioprine toxicities cannot be predicted. PMID- 17414841 TI - Update on viral hepatitis: 2006. AB - PURPOSE OF REVIEW: This is a concise review of recent developments in the field of viral hepatitis, based on publications between December 2005 and November 2006. RECENT FINDINGS: Elevated hepatitis B virus DNA levels in patients in their 40s with perinatally acquired hepatitis B virus infection increases the risk for cirrhosis and hepatocellular carcinoma. Six approved therapies are available for chronic hepatitis B. Entecavir is a potent antiviral for nucleoside-naive patients. For lamivudine resistant hepatitis B virus infection, adefovir should be added to lamivudine to reduce the risk of adefovir-resistant mutations; however, tenofovir may be a more promising alternative to adefovir. A shorter duration of treatment wth pegylated interferon and ribavirin is sufficient for genotype 2 hepatitis C infection but the benefits of extending treatment to 72 weeks for genotype 1 needs to be confirmed. Pegylated interferon monotherapy was shown to be effective in patients with hepatitis D and ribavirin provides no additional benefit. SUMMARY: New developments in the past year will help us fine tune the treatment of viral hepatitis. Even as new treatments are approved, the potential benefits of treatment should be weighed against the risk of drug resistant mutations with long-term therapy. PMID- 17414842 TI - Advancements in hepatocellular carcinoma. AB - PURPOSE OF REVIEW: This review primarily focuses on new developments in the prevention and treatment of hepatocellular carcinoma. RECENT FINDINGS: Molecular markers for tumor biology are still in development, but advances thus far are promising. The Barcelona Clinic Liver Cancer (BCLC) staging system offers the best prognostic information in patients with hepatocellular carcinoma. While surgery remains the gold standard for the treatment of hepatocellular carcinoma, new methods are emerging with greater potential response, lower risk and lower cost. Percutaneous local ablative therapy has proved very effective. Chemoemobolization therapy in hepatocellular carcinoma has been effective in selected patients with improved survival reported in a large cohort. SUMMARY: New methods for the management of hepatocellular carcinoma need to continue to evolve. This includes more focused therapies to reduce tumor development or more effective adjuvants to promote positive pre and posttransplant response. With the advent of better treatment options due to a clearer understanding of the pathogenesis of hepatocellular carcinoma, there is hope that the seemingly insurmountable burden of this disease will come under better control. PMID- 17414843 TI - Portal hypertension and its complications. AB - PURPOSE OF REVIEW: Portal hypertension is responsible for most of the complications associated with cirrhosis, specifically variceal hemorrhage, ascites and hepatic encephalopathy. Progress in understanding the pathophysiology of portal hypertension and improvements in the diagnosis and management of its complications that have occurred over the last year are discussed. RECENT FINDINGS: Endothelial dysfunction contributes to the pathogenesis of portal hypertension and may represent a novel therapeutic target. Hepatic venous pressure gradient measurements, when properly performed, are useful in the management of patients with cirrhosis. Hyponatremia in cirrhosis has prognostic value and novel aquaretic and other agents may provide alternative approaches to the management of chronic liver disease. The mechanisms for bacterial translocation in cirrhosis that predisposes patients to infectious complications, such as spontaneous bacterial peritionitis, are being explored. Adrenal insufficiency is common in septic patients with advanced cirrhosis and corticosteroids may provide a survival benefit. Pulmonary disease complicates the management of patients with advanced liver disease. SUMMARY: Significant advances continue to be made in the diagnosis and management of the complications of portal hypertension in the face of an increasing burden of chronic liver disease. PMID- 17414844 TI - Alcohol and the liver. AB - PURPOSE OF REVIEW: This review aims to acquaint the reader with advances in 2006 in the epidemiology, genetics, detection, pathogenesis and treatment of alcoholic liver disease. RECENT FINDINGS: Important discoveries have been made in pathogenesis and mechanism of disease, with great emphasis on the many pathways leading to oxidative stress, and the novel mechanism of endoplasmic reticulum stress that is proving to be important in the pathogenesis of many liver diseases. The reliability of ethyl glucuronide and other biomarkers for the detection of alcohol abuse is being better established. There have been no treatment advances for alcoholic liver disease but, on balance, steroids are still favored for carefully selected patients with alcoholic hepatitis. Many compounds tested in rodents may now be available for consideration for clinical trials. Criteria for patient selection and refusal for liver transplantation are being established but the 6 months abstinence rule still holds. SUMMARY: Insights are being made into the pathogenesis of alcoholic liver disease but safe and effective therapies for both alcoholic hepatitis and alcoholic cirrhosis have yet to be discovered. PMID- 17414845 TI - Liver transplantation: an update 2007. AB - PURPOSE OF REVIEW: Recent attention in liver transplantation has focused on equity in organ allocation and management of posttransplant complications. RECENT FINDINGS: Adoption of the model for end-stage liver disease for liver allocation has been successful in implementing a system based on medical urgency rather than waiting time. Refinements are being studied in improving the prediction of mortality and improving transplant benefit by balancing pretransplant mortality and posttransplant survival. Emerging literature is examining expansion of the current criteria for transplantation of hepatocellular carcinoma and the role of neoadjuvant therapy. Chronic renal dysfunction after liver transplantation is a source of considerable morbidity. Nephron-sparing immunosuppression regimens are emerging with encouraging results. Hepatitis C virus infection is difficult to differentiate histologically from rejection, although newer markers are being developed. Antiviral and immunosuppressive strategies for reducing the severity of hepatitis C virus recurrence are discussed. Alcohol relapse is common after liver transplant in alcoholic liver disease patients and can lead to worse outcomes. SUMMARY: Organ allocation tends to evolve under the model for end-stage liver disease with a focus on maximizing transplant benefit. Hepatitis C virus, hepatocellular carcinoma, chronic renal dysfunction and alcohol relapse are major challenges, and continued research in these areas will undoubtedly lead to better outcomes for transplant recipients. PMID- 17414846 TI - Cholangiocyte biology. AB - PURPOSE OF REVIEW: Cholangiocytes are increasingly recognized as biologically important epithelia because of the diverse array of cellular processes in which they participate. Collectively, these processes define normal function and, when disturbed, account for abnormalities that cause disease. In addition, the cholangiocyte is the target cell of diverse pathologic processes (i.e. the cholangiopathies). Advances in animal models, imaging and gene silencing have further defined the roles that cholangiocytes play in signaling, transport of water, ions and solutes, and alterations that result in cholestasis. The pace of advances in technology justifies a yearly summary to identify trends, and apprise the readership of the most significant developments in cholangiocyte biology. RECENT FINDINGS: Recent progress includes insights into the molecular mechanisms of bile secretion, the development of new experimental models, technologies, hypotheses and therapies relevant to disease. Major advances have also included the identification of novel roles for receptors and a better understanding of mechanistic pathways and biologic processes. SUMMARY: This compendium of current activities in cholangiocyte biology may promote collegial sharing and exchange of novel concepts, ideas, reagents and probes, thereby promoting positive advances in the field. PMID- 17414847 TI - Pediatric hepatobiliary disease. AB - PURPOSE OF REVIEW: This review summarizes publications in pediatric hepatobiliary disease from the past year. These studies contribute to the understanding of the epidemiology, histopathology, predictors of outcome and treatment of some important pediatric liver and biliary disorders. RECENT FINDINGS: Advances in nonalcoholic fatty liver disease, primary sclerosing cholangitis, neonatal hemochromatosis, acute liver failure (from the Pediatric Acute Liver Failure Study Group), and liver transplantation are summarized. SUMMARY: Continued investigation into these hepatobiliary disorders has the potential to significantly impact the health of children. PMID- 17414848 TI - Sclerosing cholangitis. AB - PURPOSE OF REVIEW: Primary sclerosing cholangitis is a chronic cholestatic liver disease characterized by strictures of the biliary tree complicated by cirrhosis and cholangiocarcinoma. It is immune mediated, although the precise aetiology remains unknown. RECENT FINDINGS: Research into etiopathogenesis and epidemiology, diagnosis of cholangiocarcinoma, associations with inflammatory bowel disease and autoimmune pancreatitis, and medical therapy are discussed. SUMMARY: Multiple gene polymorphisms associated with primary sclerosing cholangitis have been investigated. Common inflammatory bowel disease-associated polymorphisms do not confer any susceptibility to primary sclerosing cholangitis; the role of intercellular adhesion molecule-1 gene polymorphisms and CCR5 mutations remain unclear. Elevated IgG4 has been demonstrated in a subgroup of primary sclerosing cholangitis patients, which may indicate an overlap with autoimmune pancreatitis and possible responsiveness to steroids. Biliary brush cytology may assist in diagnosis of cholangiocarcinoma, although further clinical indicators are required. Animal studies suggest the superiority of 24 norursodeoxycholic acid over ursodeoxycholic acid in reducing histological disease progress; translational studies in humans are now required. PMID- 17414849 TI - Cholangiocarcinoma: emerging approaches to a challenging cancer. AB - PURPOSE OF REVIEW: Cholangiocarcinomas are malignancies arising from biliary tract epithelia that are associated with a poor prognosis. The diagnosis, evaluation and management of these tumors continue to be clinically challenging. In this review, we will discuss recent information that may influence the approach to the diagnosis and management of this devastating tumor. RECENT FINDINGS: Epidemiological studies are defining patient groups at increased risk of cancer in regions of low prevalence. Emerging approaches that may improve evaluation and management include the potential use of genomic and proteomic markers, cytogenetic and digital image analysis, endoscopic ultrasound with fine needle aspiration, and cholangioscopy. Adoption of aggressive surgical approaches along with the use of preoperative portal vein embolization, hepatic and caudate lobe resection and preoperative laparoscopy may improve survival. SUMMARY: Recent conceptual and technical advances will impact the clinical approach to cholangiocarcinoma and offer the promise of improving outcomes from this difficult tumor. PMID- 17414850 TI - Updates in biliary endoscopy 2006. AB - PURPOSE OF REVIEW: Endoscopic retrograde cholangiopancreatography is reserved primarily for therapeutic reasons. Recent literature continues to support therapeutic uses of the technique. This review addresses the literature in the field of biliary endoscopy for the year 2006 and is intended to assist gastroenterologists and gastrointestinal surgeons in everyday practice. RECENT FINDINGS: Endoscopic management of choledocholithiasis in gallstone pancreatitis, a newer approach in the endoscopic management of malignant biliary strictures, the broadening therapeutic indications including the use of gallbladder stenting and the performance of endoscopic retrograde cholangiopancreatography in patients with Roux-en-Y gastric bypass are discussed. Safety of the technique continues to be addressed. The risks of pancreatitis after endoscopic retrograde cholangiopancreatography as well as morbidity in the elderly are addressed. SUMMARY: Major updates in the management of biliary tract disease using biliary endoscopy are discussed over a broad range of biliary tract diseases. The literature emphasizes the broadening therapeutic role of endoscopic retrograde cholangiopancreatography as well as improvements in our understanding of risk factors for complications and the potential for their prevention. PMID- 17414851 TI - Bibliography. Current world literature. Liver. PMID- 17414852 TI - Creation of a career enhancement program for a hospital-based education and development department. AB - This article reviews the development of a career enhancement program for hospital based staff development consultants in a complex, centralized department consisting of diverse educator roles. Just as incentives for career enhancement are essential for retaining bedside nurses, it is equally vital for master's prepared educators to have a growth and recognition plan. This theory-based model can provide the needed link to achieve individual and organizational goals related to educator development and recognition. PMID- 17414854 TI - Nurses' perceptions of stress and support in the preceptor role. AB - This study was undertaken to examine whether staff nurses experience stress when they are precepting, and the sources of the stress. Another purpose was to describe preceptors' perceptions of support from others, such as coworkers and managers. A convenience sample of 65 registered nurse preceptors completed an investigator-developed questionnaire asking for their views on stress and support in the preceptor role. Most respondents (89%) reported that precepting is stressful to some degree, with the most common reasons being workload, skill level of the preceptee, organizational support, and preceptor confidence. Despite the high levels of stress reported, most subjects described adequate support as preceptors from coworkers and managers. Preceptors are critical for the training and retention of new nurses. It is important that stressors related to precepting be acknowledged and strategies be employed to reduce stress. Recommendations for support and strategies are included. PMID- 17414856 TI - An innovative approach to orient nursing students. AB - Limited resources force facilities to think beyond traditional approaches to teaching. It is vital for nursing leaders to be pioneers in the delivery of educational programs and to orient students to provide safe, competent care to clients. This research focuses on the use of a self-paced computer orientation program to facilitate the transition of nursing students into the clinical area. The data support a decrease in traditional classroom orientation time and reveal satisfaction among students. PMID- 17414857 TI - A nurse residency program for competency development using human patient simulation. AB - A new graduate registered nurse residency program integrating human patient simulation was developed at an academic medical center. The program focused on orientation through skill-based learning, critical thinking, human factors engineering, and patient safety using simulated experiences for a wide variety of high-risk, low-frequency, as well as high-frequency, commonly occurring clinical events and situations. Structured evaluations demonstrated that simulation serves as a highly effective strategy for developing competency, confidence, and readiness for entry-into-practice. It strengthened assessment and clinical skills, and enhanced nurse residents' ability to apply critical thinking to simulated patient scenarios. The time and cost of orientation decreased while recent graduate nurse satisfaction with orientation was high. PMID- 17414858 TI - A growing challenge: patient education in a diverse America. AB - Patient education is a vital part of nursing practice, but the inability to provide consistent culturally sensitive patient care to minority populations has most certainly contributed to disparities in health and healthcare. This article explores minority populations in the United States and their characteristics in relation to health and healthcare, popular cultural competence theories, and nursing school curricula, and discusses teaching strategies for developing more culturally competent nursing professionals. PMID- 17414859 TI - Staff development story. PMID- 17414860 TI - Team of teachers. PMID- 17414861 TI - Preceptorship. PMID- 17414864 TI - "Retinal Migraine" is an oxymoron. PMID- 17414865 TI - Most cases labeled as "retinal migraine" are not migraine. AB - BACKGROUND: Monocular visual loss has often been labeled "retinal migraine." Yet there is reason to believe that many such cases do not meet the criteria set out by the International Headache Society (IHS), which defines "retinal migraine" as attacks of fully reversible monocular visual disturbance associated with migraine headache and a normal neuro-ophthalmic examination between attacks. METHODS: We performed a literature search of articles mentioning "retinal migraine," "anterior visual pathway migraine," "monocular migraine," "ocular migraine," "retinal vasospasm," "transient monocular visual loss," and "retinal spreading depression" using Medline and older textbooks. We applied the IHS criteria for retinal migraine to all cases so labeled. To be included as definite retinal migraine, patients were required to have had at least two episodes of transient monocular visual loss associated with, or followed by, a headache with migrainous features. RESULTS: Only 16 patients with transient monocular visual loss had clinical manifestations consistent with retinal migraine. Only 5 of these patients met the IHS criteria for definite retinal migraine. No patient with permanent visual loss met the IHS criteria for retinal migraine. CONCLUSIONS: Definite retinal migraine, as defined by the IHS criteria, is an exceedingly rare cause of transient monocular visual loss. There are no convincing reports of permanent monocular visual loss associated with migraine. Most cases of transient monocular visual loss diagnosed as retinal migraine would more properly be diagnosed as "presumed retinal vasospasm." PMID- 17414866 TI - Wall-eyed bilateral internuclear ophthalmoplegia from lesions at different levels in the brainstem. AB - Wall-eyed bilateral internuclear ophthalmoplegia (WEBINO) is a rarely reported syndrome. There is dispute about whether WEBINO is caused by a pontine or a midbrain lesion and whether the medial rectus subnuclei are implicated. In a study of the clinical and imaging features of four patients with WEBINO, we found that that three of four lesions involved the midbrain but not necessarily the medial rectus subnuclei. PMID- 17414867 TI - Optic neuropathy associated with periostitis in relapsing polychondritis. AB - Optic neuropathy is an uncommon manifestation of relapsing polychondritis (RPC), a rare systemic disease affecting cartilaginous and proteoglycan-rich structures. The optic neuropathy has been attributed to ischemia, intrinsic inflammation of the optic nerve, or spread of inflammation to the nerve from adjacent intraconal orbital tissues. We report a case of recurrent corticosteroid-responsive optic neuropathy in which MRI did not show ocular, optic nerve, or intraconal orbital abnormalities but did show periosteal thickening and enhancement in the apical orbit and adjacent intracranial space consistent with periostitis. The periostitis, which is a manifestation of a systemic vasculitis or an autoimmune reaction to progenitors of cartilage, probably caused the optic neuropathy by compression or inflammation. It is important to recognize this mechanism of optic neuropathy as its imaging features may be a subtle yet critical clue to an underlying systemic condition that can be life-threatening if not properly managed. PMID- 17414868 TI - Ocular motor disorders in mitochondrial encephalopathy with lactic acid and stroke-like episodes with the 3271 (T-C) point mutation in mitochondrial DNA. AB - BACKGROUND: Ocular motor function can provide insights into areas of dysfunction within the nervous system. There are no published eye movement recordings in patients with mitochondrial encephalopathy with lactic acid and stroke-like episodes (MELAS). Our purpose in this study was to analyze the ocular motor features of a family with MELAS with a (T-C) mutation at nucleotide position 3271 in the mitochondrial tRNA-Leu gene. METHODS: The search coil method was used to record visually-guided saccades, antisaccades, and triangular pursuit tasks in the horizontal and vertical planes in three patients in a Japanese family with MELAS. RESULTS: The patients showed saccadic dysmetria and prolonged saccadic reaction times, deficits in the ability to suppress reflex eye movements, and increased reaction time during antisaccades, downbeat nystagmus, square wave jerks, and impairment in pursuit. CONCLUSIONS: On the basis of eye movement recordings, patients with MELAS have frontal cortex as well as cerebellar dysfunction. PMID- 17414869 TI - Painful sixth cranial nerve palsy caused by a malignant trigeminal nerve sheath tumor. AB - A 17-year-old woman developed a sixth cranial nerve palsy from a malignant peripheral nerve sheath tumor of the trigeminal nerve. This case is unusual in that the principal symptom was diplopia stemming from a sixth cranial nerve palsy. Pain was mild, and trigeminal function was preserved. Imaging evidence of rapid growth of the cavernous sinus mass gave rise to an initial impression that the cause might be inflammatory. Treatment with gamma knife stereotactic radiosurgery produced some improvement in sixth cranial nerve function and reduction in tumor size over a follow-up period of 9 months. PMID- 17414870 TI - Modified Lundie loops improve apraxia of eyelid opening. AB - BACKGROUND: Current treatments are unsatisfactory for improving apraxia of eyelid opening, defined as a delay or inability to open closed eyelids voluntarily in the presence of intact motor pathways. METHODS: Improvement in functional health was assessed using the Blepharospasm Disability Scale (BDS) in five consecutive patients with apraxia of eyelid opening treated with wire loops affixed behind ordinary spectacles (Lundie loops) and modified to provide pressure on the brow as a stimulus to keep the eyelids elevated. RESULTS: All five patients showed improvement in BDS scores. The mean percentage of normal activity of the study population improved from 25% to 37.6%. Outdoor activities were not significantly altered with the use of the device. CONCLUSIONS: Modified Lundie loops appear to be helpful in improving the functional health of patients with eyelid apraxia. These results will need to be verified in larger trials. PMID- 17414871 TI - Vogt-Koyanagi-Harada syndrome in a case of multiple sclerosis. AB - A 34-year-old woman in whom multiple sclerosis (MS) had been diagnosed 2 years earlier developed progressive bilateral visual loss associated with peripapillary exudative retinal detachment and other features of Vogt-Koyanagi-Harada (VKH) syndrome. She was treated with corticosteroid pulses and interferon beta-1A with visual acuity improvement and resolution of the retinal detachment. This is the first reported case of VKH syndrome in a patient with MS. The combination of VKH syndrome and MS suggests a common autoimmune pathogenesis. PMID- 17414872 TI - Mitochondrial ophthalmoplegia with fatigable weakness and elevated acetylcholine receptor antibody. AB - A 25-year-old man with chronically progressive ptosis and bilateral ophthalmoplegia displayed fatigability and fluctuation of ptosis, an abnormal single-fiber electromyogram, and a markedly elevated acetylcholine receptor antibody level. Yet a muscle biopsy showed clear evidence of a mitochondrial cytopathy, and the clinical features did not improve after treatment with prednisone. This case emphasizes the difficulty in differentiating mitochondrial cytopathy from myasthenia gravis and points out that elevated acetylcholine receptor antibody levels may occur in nonmyasthenic conditions. PMID- 17414873 TI - Accelerated growth of an orbital schwannoma during pregnancy. AB - An incidentally diagnosed unilateral orbital mass in a 34-year-old woman grew at an accelerated rate during pregnancy with deterioration of visual function. Removed early after delivery by a transcranial approach, the tumor was histologically diagnosed as a richly vascularized cystic schwannoma containing red blood cell components. The accelerated tumor growth was attributed to intratumoral hemorrhage. Although acoustic schwannomas have been reported to enlarge during pregnancy, this phenomenon has only been described once for orbital schwannomas. Because this type of tumor may enlarge during pregnancy and threaten visual function, surgery should not be delayed. PMID- 17414874 TI - Transorbital intracranial penetrating injury from impaling on an earpick. AB - An inebriated 86-year-old man impaled himself on a wooden earpick that penetrated through the superior orbital fissure into the prepontine cistern. The patient underwent surgery immediately by a lateral suboccipital approach, and the earpick was pulled out through the wound with control of hemorrhage from the cavernous sinus. He survived this event with no neurologic deficits apart from complete ipsilateral ophthalmoplegia and ptosis. Prompt imaging and surgical intervention allowing direct visualization of the foreign body and prevention of intracranial complications are part of proper management of this problem. PMID- 17414875 TI - Papilledema in obstructive hydrocephalus caused by giant cell astrocytoma of tuberous sclerosis. AB - A 5-year-old girl with progressive hemiparesis and headache was found by brain imaging to have a large tumor centered at the foramen of Monro, blocking cerebrospinal outflow and producing massive lateral ventriculomegaly. Total excision of the mass led to a pathologic diagnosis of giant cell astrocytoma. Dermatologic abnormalities had been detected shortly after birth but were unexplained. Abdominal imaging disclosed renal cysts, and ophthalmologic examination disclosed papilledema and retinal plaques. On this basis, a diagnosis of tuberous sclerosis (TS) was finally made. Two months after surgery, papilledema had resolved, and visual function appeared to be normal. Although the patient apparently escaped visual loss, other reports affirm that giant cell astrocytoma, a common tumor in TS, may go undetected for long enough to produce irreversible optic neuropathy from chronic papilledema. Because patients with TS may not report visual loss, they should undergo periodic ophthalmologic screening. PMID- 17414876 TI - Susac syndrome in a patient with hepatitis C. AB - A 38-year-old woman seropositive for hepatitis C developed headache, sensorineural hearing loss, encephalopathy, and retinal arteriolar occlusions. Brain MRI showed signal abnormalities in the basal ganglia and corpus callosum. These features are consistent with Susac syndrome, a multifocal central nervous system disorder of uncertain etiology. This is the first reported case of Susac syndrome in a patient with hepatitis C. PMID- 17414877 TI - Rethinking neuromyelitis optica (Devic disease). AB - Neuromyelitis optica (NMO), or Devic disease, has been distinguished from multiple sclerosis (MS) by the presence of optic neuritis that is usually bilateral, simultaneous, and often severe, myelopathic findings accompanied by longitudinally extensive spinal cord imaging abnormalities, no brain imaging abnormalities typical of MS, and often rapid progression to debility and even death. Researchers at the Mayo Clinic have identified an immunoglobulin marker of NMO (the "NMO antibody") that binds selectively to the aquaphorin-4 water channel and may play a causative role. This marker has been found in Japanese patients with opticospinal MS, prompting the suggestion that NMO and Japanese opticospinal MS are the same disorder. The NMO antibody, which predicts frequent relapse of myelopathy and optic neuritis, is also found in patients with lupus erythematosus and Sjogren syndrome who also have severe optic neuritis and longitudinally extensive myelitis. Because this antibody is also found in patients with optic neuritis and myelitis who have brain signal abnormalities atypical of MS, the diagnosis of NMO has been revised to allow inclusion of these brain imaging abnormalities. Proper distinction of NMO from MS is important because the two disorders may respond differently to immune modulatory therapy. PMID- 17414878 TI - Simmons Lessell: the gaon of neuro-ophthalmology. PMID- 17414880 TI - Fluctuating ptosis, diplopia, and normal pupils with intracavernous aneurysm. PMID- 17414881 TI - Retinal migraine. PMID- 17414882 TI - Amiodarone and ischemic optic neuropathy. PMID- 17414883 TI - The role of FDG PET in the management of lymphoma: what is the evidence base? AB - [18F]Fluorodeoxyglucose positron emission tomography (18F-FDG PET) is playing an increasing role in the management of both Hodgkin and non-Hodgkin lymphoma, offering potential advantages in the accuracy of disease assessment at a number of points in the management pathway. This review evaluates the current level of confidence in the use of PET technology in (1) initial staging, (2) the assessment of early response to chemotherapy, (3) the assessment of residual masses at completion of initial treatment, (4) follow-up, and (5) radiotherapy planning. PMID- 17414884 TI - The role of FDG PET in the management of lymphoma: practical guidelines. AB - Practical guidelines for the use of FDG PET in the management of lymphoma are given, based on the evidence presented in the previous article. A statement and recommendations are given where appropriate. The recommendations are summarized at the end of the paper. PMID- 17414885 TI - 68Ga-DOTATATE PET in neuroectodermal tumours: first experience. AB - BACKGROUND AND AIM: Phaeochromocytoma is initially imaged with computed tomography (CT) or magnetic resonance imaging (MRI) but functional imaging is commonly needed to assess disease activity, the presence of metastasis and response to therapy. Traditionally, this is done with 123I -MIBG with good sensitivity and specificity. However, spatial resolution remains limited even with SPECT. We aimed to assess the utility of a new somatostatin analogue PET tracer, 68Ga-DOTATATE in the management of phaeochromocytoma. METHODS: We retrospectively reviewed five patients with malignant phaeochromocytoma who underwent imaging with CT and 123I-MIBG and compared the results with those of PET imaging using 68Ga-DOTATATE. Blinded analysis of the numbers and extent of lesions were done for all imaging modality. RESULTS: Two patients had negative 123I-MIBG and positive 68Ga-DOTATATE scans. One had a weakly positive 123I-MIBG and a strongly positive 68Ga-DOTATATE scan. One had a positive 123I-MIBG and positive 68Ga-DOTATATE scans. The fifth patient was negative to all imaging including CT. 68Ga-DOTATATE showed more lesions with higher uptake and better resolution compared to 123I-MIBG. CONCLUSION: The findings in our small group of patients demonstrate the value of somatostatin receptor PET imaging in malignant phaeochromocytoma. In lesions with no or low MIBG uptake, the next investigation of choice should be PET imaging with 68Ga-DOTATATE, in view to therapy with Y labelled DOTATATE. PMID- 17414886 TI - In search of an unknown primary tumour presenting with cervical metastases: performance of hybrid FDG-PET-CT. AB - OBJECTIVES: In patients with cervical lymph node metastases from unknown primary tumour (UPT), the primary tumour is frequently localized in the head and neck area. Because the detection of the primary tumour is of importance to optimize the patient's management and allows a targeted therapy, the performances of hybrid positron emission tomography-computed tomography (PET-CT) using fluorodeoxyglucose (FDG) were evaluated in a retrospective study. METHODS: Thirty eight consecutive patients with cervical lymph node metastases, and in whom the primary was not detected by the comprehensive diagnostic work-up including endoscopy and conventional imaging methods, were referred for a PET-CT scan. RESULTS: PET-CT was positive with an increased FDG focal uptake suggesting the potential primary site in 68% of patients (26/38), which guided the biopsies during a second rigid panendoscopy in 17 of these 26 patients: 13 primary tumours were then histologically proven. PET-CT showed distant lesions in three patients. It had treatment-related implications in 23/38 patients (60%), consisting of modification of radiation planning, surgery or abstention from surgery. CONCLUSION: Hybrid FDG-PET-CT is helpful for the detection of a potential head and neck primary tumour. Furthermore, hybrid FDG-PET-CT has the ability to diagnose occult or distant second tumour and metastatic disease and modify patient management. PMID- 17414887 TI - The value of quantifying 18F-FDG uptake in thyroid nodules found incidentally on whole-body PET-CT. AB - OBJECTIVE: To determine if quantification of [18F]fluorodeoxyglucose (18F-FDG) uptake in a thyroid nodule found incidentally on whole-body 18F-FDG positron emission tomography-computed tomography (PET-CT) can be used to discriminate between malignant and benign aetiology. METHODS: A retrospective review of all patients with focally high uptake in the thyroid as an incidental finding on 18F FDG PET-CT from May 2003 through May 2006. The uptake in the nodules was quantified using the maximum standardized uptake value (SUVmax). The aetiology was determined by cytology and/or ultrasound, or on histopathology. RESULTS: Incidental focally high uptake was found in 79/7347 patients (1.1%). In 31/48 patients with adequate follow-up, a benign aetiology was determined. Median SUVmax for the benign group was 5.6, range 2.5-53. Malignancy was confirmed in 15/48 patients. The malignancies were papillary thyroid carcinoma in 12, metastasis from squamous cell carcinoma in one, and lymphoma in two. Median SUVmax for the malignant lesions was 6.4, range 3.5-16. Cytology suspicious for follicular carcinoma was found in 2/48 patients. No statistical difference (P=0.12) was found among the SUVmax between the benign and malignant groups. CONCLUSION: Focally high uptake of 18F-FDG in the thyroid as an incidental finding occurred in 1.1% of the patients. Malignancy was confirmed or was suspicious in 17/48 (35%) of the patients that had adequate follow-up. There was no significant difference in the SUVmax between benign and malignant nodules. PMID- 17414888 TI - Influence of the different biokinetics of sestamibi and tetrofosmin on the interpretation of myocardial perfusion imaging in daily practice. AB - AIM: Digestive activity can interfere with the interpretation of myocardial perfusion single photon emission computed tomography using sestamibi or tetrofosmin. Compared with sestamibi, the liver clearance of tetrofosmin is more rapid, but its absolute cardiac uptake is lower. In this study, the activity of sestamibi and tetrofosmin was quantified after exercise or pharmacological stress and at rest to objectify the biokinetic differences and to evaluate whether there is a correlation between quantitative measurements and the visual assessment of image quality. METHODS: Left ventricular activity and five ratios (R1-R5) of cardiac to adjacent extra-cardiac activity were quantified in 204 sestamibi (68 exercise stress/56 pharmacological stress/80 rest) and 221 tetrofosmin (67 exercise stress/59 pharmacological stress/95 rest) studies. Image quality was assessed by a three-point score (1, good; 2, moderate; 3, poor) and correlated with the heart to left supra-diaphragmatic region (R1) and heart to right supra diaphragmatic region (R2) ratios. RESULTS: The mean left ventricular activity was higher for sestamibi, especially at rest (sestamibi, 0.21+/-0.05 counts/pixel/injected MBq; tetrofosmin, 0.16+/-0.042 counts/pixel/injected MBq; P<0.001). By contrast, most ratios were higher with tetrofosmin, particularly for the exercise stress and rest studies. Using the three-point quality scoring, more sestamibi than tetrofosmin studies were scored as 3 (12.2% versus 6.3%), also particularly for the exercise stress and rest studies. A highly significant relationship was found between decreasing R1 and R2 and an increasing quality score, regardless of the radiopharmaceutical used (P values between 0.02 and <0.001). CONCLUSIONS: Despite a lower cardiac uptake, the more rapid liver clearance of tetrofosmin than sestamibi significantly improves the ratios of cardiac to digestive activity, especially after exercise or at rest. These quantitative differences in biokinetics result in less poor scans with tetrofosmin in daily practice. PMID- 17414889 TI - Electrocardiograph-gated single photon emission computed tomography radionuclide angiography presents good interstudy reproducibility for the quantification of global systolic right ventricular function. AB - BACKGROUND: Electrocardiograph-gated single photon emission computed tomography (SPECT) radionuclide angiography provides accurate measurement of right ventricular ejection fraction and end-diastolic and end-systolic volumes. AIM: In this study, we report the interstudy precision and reliability of SPECT radionuclide angiography for the measurement of global systolic right ventricular function using two, three-dimensional volume processing methods (SPECT-QBS, SPECT 35%). These were compared with equilibrium planar radionuclide angiography. METHODS: Ten patients with chronic coronary artery disease having two SPECT and planar radionuclide angiography acquisitions were included. RESULTS: For the right ventricular ejection fraction, end-diastolic volume and end-systolic volume, the interstudy precision and reliability were better with SPECT-35% than with SPECT-QBS. The sample sizes needed to objectify a change in right ventricular volumes or ejection fraction were lower with SPECT-35% than with SPECT-QBS. The interstudy precision and reliability of SPECT-35% and SPECT-QBS for the right ventricle were better than those of equilibrium planar radionuclide angiography, but poorer than those previously reported for the left ventricle with SPECT radionuclide angiography on the same population. CONCLUSION: SPECT-35% and SPECT-QBS present good interstudy precision and reliability for right ventricular function, with the results favouring the use of SPECT-35%. The results are better than those of equilibrium planar radionuclide angiography, but poorer than those previously reported for the left ventricle with SPECT radionuclide angiography. They need to be confirmed in a larger population. PMID- 17414890 TI - The relation between striatal dopamine D2/D3 receptor availability and sleep quality in healthy adults. AB - PURPOSE: Increasing evidence, primarily from animal studies and patients with compromised neurotransmitter systems, indicates a possibly important role for dopamine in modulating sleep. We therefore conducted this study to explore the relation between sleep and dopamine in healthy adults. METHODS: We used the Pittsburgh Sleep Quality Index (PSQI) to assess sleep quality. Higher PSQI scores indicate a lower quality of sleep. Striatal dopamine D2/D3 receptor availability was determined using [123I]iodo-benzamide (IBZM) SPECT. Fifty-five healthy volunteers (32 men, 23 women; mean age, 36.7+/-12.1 years), including 25 good sleepers and 30 poor sleepers, were recruited. We analysed the correlation between the PSQI and D2/D3 receptor availability in good and poor sleepers based on Pearson's product-moment after removing the effects of gender and age. We also analysed differences in D2/D3 receptor availability between good and poor sleepers. RESULTS: In poor sleepers, there was no statistically significant relationship between the global, individual components of the PSQI score and D2/D3 receptor availability. However, in good sleepers, the score of the sleep duration component was significantly negatively correlated with D2/D3 receptor availability in the caudate. There was no significant difference in D2/D3 receptor availability between good and poor sleepers. CONCLUSION: Our findings demonstrate that healthy good sleepers with higher D2/D3 receptor availability in the caudate sleep longer. Poor sleep in healthy subjects might be not primarily related to the dopaminergic system. PMID- 17414891 TI - Development of formulae for accurate measurement of the glomerular filtration rate by renal dynamic imaging. AB - AIM: Currently, the widely adopted renal dynamic imaging in clinical practice uses Gates' method to calculate the glomerular filtration rate (GFR), but many researchers have proven that Gates' method may result in bias. Thus, this article explores alternative improved formulae to calculate GFR by renal dynamic imaging. METHODS: Three hundred and sixty-seven patients were selected and their GFR values were measured using renal dynamic imaging and the two-plasma method with 99mTc-diethylenetriaminepentaacetic acid (99mTc-DTPA) as the imaging agent. With the two-plasma GFR as reference value, two equations were obtained from linear and non-linear regression analyses between the renal uptake percentage and two plasma GFR. The 367 patients were divided into two random groups, with the first group used to derive the regression formulae and the second to verify the formulae. Finally, all patients were studied to derive the formulae to calculate GFR. The comparison of our formulae with the commonly used Gates' formula was conducted by the Bland-Altman method. RESULTS: The linear and non-linear GFR formulae were as follows: GFR (ml/min/1.73 m2)=(631.633 x renal uptake percentage - 2.040) x 1.73/BSA (BSA, body surface area) and GFR (ml/min/1.73 m2)=(-1996.585 x renal uptake percentage2 + 1013.526 x renal uptake percentage - 12.739) x 1.73/BSA, respectively. The biases of the GFR values calculated using the linear and non-linear formulae and Gates' formula relative to the two-plasma GFR were 2.5 +/- 19.1 ml/min/1.73 m2, -2.0 +/- 19.3 ml/min/1.73 m2 and 3.4 +/- 19.4 ml/min/1.73 m2, respectively. CONCLUSIONS: The GFR values calculated using our new formulae correlate better with the reference GFR value than does GFR calculated by Gates' formula, and the GFR values measured using the non-linear formula are more accurate than those obtained using the linear formula. PMID- 17414892 TI - Can oncology patients' central venous catheters be used for isotope assessment of glomerular filtration rate? An in-vitro study. AB - BACKGROUND: Isotope assessment of glomerular filtration rate (GFR) is frequently performed in patients with central venous catheters (CVCs). Use of the CVCs for administration of tracer and subsequent blood sampling would be less distressing for patients (particularly paediatric) and would reduce the frequency of failed samples due to poor venous access. However, the GFR test is quantitative and could be affected by incomplete tracer delivery due to adhesion to the CVC and also by contamination of blood samples due to adhered tracer leaching back into the sampled blood as it passes through the CVC. AIM: This in vitro study aimed to quantify the effects on GFR assessment of tracer adhesion and leaching, in single lumen and dual-lumen CVCs. METHOD: New and clinically used single-lumen CVCs were injected with tracer (99mTc-DTPA and 51Cr-EDTA) and then flushed repeatedly with saline. The outflows were assayed in a gamma counter and, where possible, the CVCs were imaged on a gamma camera to take snap shots of tracer movement throughout a GFR assessment. In a separate experiment, a phantom patient was used to compare blood sampling through a dual lumen CVC with peripheral sampling. RESULT AND CONCLUSION: A CVC successfully delivers >99% of tracer. Subsequent blood samples can be taken through the other lumen of a dual-lumen CVC but not through a single-lumen as this significantly alters the GFR result due to contamination. PMID- 17414893 TI - Re: training staff for the delivery of PET/CT services in the UK. PMID- 17414894 TI - Passive incorporation of radioisotopes and airport security. PMID- 17414895 TI - Is there life in MARS? PMID- 17414896 TI - Three-dimensional kinetics of the lumbar spine and hips in low back pain patients during sit-to-stand and stand-to-sit. AB - STUDY DESIGN: Experimental, biomechanical study to determine the kinetics of the lumbar spine and hips during sit-to-stand and stand-to-sit. OBJECTIVE: To investigate the effects of back pain, with and without limitation in straight leg raise, on the joint moment and power of the lumbar and hips during sit-to-stand and stand-to-sit. SUMMARY OF BACKGROUND DATA: Movements of the lumbar spine and hips, and their coordination have been reported to be affected by the presence of low back pain (LBP), especially in those with a positive straight leg raise. However, the literature has no information concerning moment and power characteristics of the lumbar spine and hips during sit-to-stand and stand-to-sit in such patients. METHODS: Twenty asymptomatic subjects, 20 LBP patients, and 20 patients with LBP and a positive straight leg raise sign were requested to perform the sit-to-stand and stand-to-sit activities. Electromagnetic sensors were attached to the body segments to measure their kinematics while 2 nonconductive force plates gathered ground reaction force data. Biomechanical models were used to determine the muscle moments and power at the lumbosacral (L5/S1) joint and hips. RESULTS: Muscle moments acting at the lumbar spine and hip in the sagittal plane were found to decrease in subjects with LBP, but there were significant increases in moments in other planes of motion. The power patterns of the spine and hips were also significantly altered, particularly in subjects with a positive straight leg raise sign. CONCLUSIONS: Back pain subjects exhibit compensatory movements and altered load sharing strategies during the sit to-stand and stand-to-sit activities. Exercise therapy should take account of these changes so that the normal kinematic and kinetic characteristics of the spine and hips can be restored. PMID- 17414897 TI - Fluoroscopic video to identify aberrant lumbar motion. AB - STUDY DESIGN: A prospective, case-control design. OBJECTIVES: To develop a kinematic model that characterizes frequently observed movement patterns in patients with low back pain (LBP). SUMMARY OF BACKGROUND DATA: Understanding arthrokinematics of lumbar motion in those with LBP may provide further understanding of this condition. METHODS: Digital fluoroscopic video (DFV) was used to quantify the magnitude and rate of attainment of sagittal plane intersegmental angular and linear displacement from 20 individuals with LBP and 20 healthy control subjects during lumbar flexion and extension. Three fellowship trained spine surgeons subsequently qualitatively analyzed the DFVs to determine normality of movement. Final classification was based on agreement between their symptom and motion status (11 with LBP and aberrant motion and 14 healthy controls without aberrant motion). Independent t tests, receiver operator characteristic curves, and accuracy statistics were calculated to determine the most parsimonious set of kinematic variables able to distinguish patients with LBP. RESULTS: Eight kinematic variables had a positive likelihood ratio > or = 2.5 and entered the model. Six of the variables described a disruption in the rate of attainment of angular or linear displacement during midrange postures. When 4 or more of these variables were present, the positive likelihood ratio was 14.0 (confidence interval 3.2-78.5), resulting in accurately identifying 96% of participants. CONCLUSIONS: DFV was useful for discriminating between individuals with and without LBP based on kinematic parameters. Disruptions in how the motion occurred during midrange motions were more diagnostic for LBP than range of motion variables. Cross validation of the model is required. PMID- 17414898 TI - Analysis of pedicle and translaminar facet fixation in a multisegment interbody fusion model. AB - STUDY DESIGN: This is a biomechanical study. We compared pedicle screws and translaminar screws in the setting of multisegment interbody fusions. OBJECTIVES: We investigated the significance of the middle segment pedicle screws in a 2 level interbody fusion model and examined translaminar screw fixation as an alternative to pedicle screw fixation in this 2-level model. SUMMARY OF BACKGROUND DATA: Surgical treatment of disc disease focuses on restoration of normal disc height and restriction of abnormal motion. Interbody fusion significantly reduces motion and restores disc space height. Combined anterior and posterior fusion improves fusion rates. METHODS: Human cadavers were tested for range of motion (ROM) and stiffness. Each specimen underwent a 2-level interbody fusion and posterior fixation. There were 3 types of posterior fixation: pedicle screws at 3 levels, pedicle screws at 2 levels, and translaminar facet screws. Biomechanical testing was repeated for each group. RESULTS: The overall ROM decreased after the interbody fusion (P < 0.05). There were no significant differences in construct stiffness for torsion, but there were significant differences when comparing intact to the 3 groups for both flexion and extension (P < 0.05). ROM at L2 and L4 was significantly less for all groups in comparison to the intact condition (P < 0.05). CONCLUSIONS: This study supports the omission of the middle segment screws in a 2-level interbody fusion. In addition, translaminar screws are similar to pedicle fixation in a 2-level interbody lumbar fusion. PMID- 17414899 TI - Transmaxillary and transmandibular approach to a C1 chordoma. AB - STUDY DESIGN: Case report. OBJECTIVES: To demonstrate the efficacy of a transmaxillary and transmandibular approach in achieving a wide view and the aggressive resection of a retropharyngeal chordoma originating from C1. SUMMARY OF BACKGROUND DATA: Although aggressive surgical resection has been recommended for the treatment of chordomas, wide exposure of the tumors in the upper cervical region is a challenge. METHODS: A 19-year-old man presented with a large ossified retropharyngeal chordoma (6 cm in diameter) originating from the right side of the anterior arch of C1, and extending from the clivus to the C2/3 intervertebral disc level in the sagittal plane. A posterior occipitocervical (O-C3) fusion with an iliac bone graft was first performed. Ten days after the fusion, the tumor was resected using a mandible and tongue midsplitting approach combined with a Le Fort I (transmaxillary) osteotomy, which allowed us to expose the entire tumor. The tumor was hard and immovable because of ossification. The main part of the tumor was resected from the anterior arch of C1, and then the C1 anterior arch was resected en bloc. The retropharyngeal wall was reconstructed using a vascularized radial forearm flap. Radiation therapy (60 Gy) was performed after surgery. RESULTS: No local recurrence or metastasis was observed 3 years after the operation. The patient had no complaints and has returned to his previous job as a manual laborer. CONCLUSIONS: A transmaxillary and transmandibular approach allowed us to obtain a complete view of a large immovable chordoma located ventral to the upper cervical spine. This enabled us to resect totally the tumor into 2 pieces without major complications or sequelae. This approach is useful for the resection of large tumors located in the median upper cervical spine. PMID- 17414900 TI - Long-segment fusion of the thoracolumbar spine in conjunction with a motion preserving artificial disc replacement: case report and review of the literature. AB - STUDY DESIGN: Case report. OBJECTIVE: We reviewed the case of a 44-year-old woman who underwent long-segment fusion and an artificial disc replacement. SUMMARY OF BACKGROUND DATA: There have been many reported advantages and disadvantages of stopping the fusion at L5, with the theoretical benefits being preserved motion, shorter operative time, allowing the remaining disc to compensate for curve correction cephalad in the lumbar spine, and a decreased likelihood for the development of a pseudarthrosis at that distal level. METHODS: As the issue of the fate of the L5-S1 motion segment continues to be debated, we present the case of a medium-segment thoracolumbar fusion carried down to the L4 stable vertebra, an intervening healthy L4-L5 disc space, with the placement of an artificial disc arthroplasty at the L5-S1 level for a degenerative and discographically positive pain generator. RESULTS: At 2-year follow-up, her L5-S1 artificial disc replacement level shows 11 degrees range of motion and consolidated fusion from T12 to L4 with complete resolution of her axial back pain. Her T12-L4 construct is stable, and the L4-L5 level is unaffected at the latest follow-up. Her clinical outcome has been excellent with her return to a very active lifestyle. CONCLUSION: Artificial disc replacement below a long-segment fusion is a viable alternative to performing fusion to additional motion segments. PMID- 17414902 TI - Effectiveness of a back pain prevention program: a cluster randomized controlled trial in an occupational setting. AB - STUDY DESIGN: A cluster randomized controlled trial and economic evaluation with a 12-month follow-up and with work department as the unit of randomization. OBJECTIVE: To evaluate the effectiveness of a prevention program for low back pain (LBP) in an occupational setting with an economic evaluation. SUMMARY OF BACKGROUND DATA: LBP accounts for high economic costs in Western societies. Little is known on the effectiveness and related costs and savings of prevention programs for LBP. METHODS: The study population consisted of workers in physically demanding jobs from 9 large companies located throughout The Netherlands. In each company, 2 comparable work units were randomly allocated, resulting in 18 clusters with 258 workers assigned to the intervention group and 231 workers to the control group. RESULTS: Results in our study did not show significant differences in effects or costs savings of the program. Indirect costs related to work absence and productivity losses accounted for the majority (84%) of total costs due to LBP. CONCLUSIONS: This study provides no evidence for the adoption of this worksite prevention program for LBP. PMID- 17414903 TI - Efficacy of mesenchymal stem cell enriched grafts in an ovine posterolateral lumbar spine model. AB - STUDY DESIGN: Four groups of 6 animals underwent single-level noninstrumented posterolateral lumbar fusion (PLF) with one of the following grafts: 1) autograft, 2) cell-enriched beta-tricalcium phosphate (TCP), 3) TCP with whole bone marrow, and 4) TCP alone. Plain radiographs were taken after surgery and at death, 6 months after surgery. Explanted spine segments were analyzed by manual palpation, micro-CT, and histology. OBJECTIVE: A sheep spine fusion study was undertaken to evaluate the healing performance of a TCP graft enriched with osteoprogenitor cells using Selective Cell Retention technology (SCR), compared with autograft, TCP with whole bone marrow, and TCP alone. SUMMARY OF BACKGROUND DATA: Improved bone healing with previously demonstrated using grafts enriched in osteoprogenitor cells. METHODS: Cell-enriched grafts were obtained by processing 30 mL of bone marrow through 10 mL of TCP. TCP was also used either saturated with bone marrow or alone. RESULTS: At 6 months, 33% of the SCR-enriched TCP and 25% of the autograft sites were fused, compared with 8% of the TCP plus whole bone marrow and 0% of the TCP alone. Histology of fused samples showed denser bone formation in the SCR-enriched TCP grafts than in the autograft sites. CONCLUSIONS: The use of SCR-enriched TCP and autograft resulted in similar fusion rates in an ovine posterolateral noninstrumented lumbar spine fusion model. PMID- 17414905 TI - Comparative analysis of serum proteomes to discover biomarkers for ossification of the posterior longitudinal ligament. AB - STUDY DESIGN: Serum proteomes from normal subjects and the patients with ossification of the posterior longitudinal ligament (OPLL) were analyzed by using proteomics. OBJECTIVES: To identify novel serologic biomarkers for diagnosing OPLL. SUMMARY OF BACKGROUND DATA: OPLL can compress the spinal cord, and special planning is required for surgeries that are done from the front of the cervical spine. However, the definitive serologic biomarkers for OPLL are still unclear. METHODS: The 2-dimensional electrophoresis patterns of sera from OPLL patients and normal subjects were compared. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and electrospray ionization quadruple time-of-flight mass spectrometry. RESULTS: Nine spots that were differentially expressed in the sera of OPLL patients were found and were identified. PRO2675, human serum albumin in a complex with myristic acid and tri-iodobenzoic acid, an unknown protein, chain B of the crystal structure of deoxy-human hemoglobin beta6, pro-apolipoprotein, ALB protein, retinol binding protein and chain A of human serum albumin mutant R218h complexed with thyroxine (3,3',5,5', tetraiodo-L-thyronine), were up regulated in the sera of OPLL patients, whereas alpha1-microglobulin/bikunin precursor was down-regulated. CONCLUSIONS: These proteins could be used as diagnostic biomarkers of OPLL. PMID- 17414906 TI - The mutant guppy syndrome curveback as a model for human heritable spinal curvature. AB - STUDY DESIGN: This study investigated the morphology, pathogenesis, and inheritance of idiopathic-like spinal curvature in the guppy syndrome, curveback. OBJECTIVE: To determine whether curveback could be applied as a model for the primary factors that contribute to heritable spinal curvature in humans, specifically, the etiopathogenesis of human familial idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Although a genetic basis is accepted, phenotypic complexity and the lack of an animal model with noninduced curvature have made identification of idiopathic scoliosis etiology difficult. It is well established that humans and fish share many genes with similar tissue and temporal expression characteristics, and comparisons between human and fish genomes have proven to be valuable for understanding the genetics of diseases affecting humans. METHODS: The curveback lineage of guppies was constructed from a single curved male crossed to a normal female. Offspring (103) from the original cross were scored from birth until death for the presence and magnitude of spinal curvature. Genetic architecture was investigated through selective inbreeding, analysis of the distribution of curve magnitude in the mature population, and assessment of curve dynamics during development. Computed tomography assessed vertebral detail. RESULTS: Computed tomography reveals that vertebral breakage or fusion is not associated with the curveback syndrome. Inbreeding demonstrates a strong genetic influence on curveback, and the distribution of curve magnitude among adult fish suggests polygenic inheritance. There is a female bias for curves of high magnitude and curves that resolve before maturity. There is developmental variability for the age of curve onset, curve progression, and final curve magnitude. CONCLUSIONS: Observed parallels between the curveback syndrome and human idiopathic scoliosis suggest that the guppy model is an unexploited resource for the identification of primary etiological factors involved in curvature. As models for biomedical research, teleosts offer great potential regarding spinal stability and deformity. PMID- 17414907 TI - Sterilization and strength of 70/30 polylactide cages: e-beam versus ethylene oxide. AB - STUDY DESIGN: In vitro and in vivo studies on the degradation of 70/30 poly(L,DL lactide) (PLDLLA) cages. OBJECTIVE: To evaluate the effect of e-beam and ethylene oxide sterilization on degradation and strength. SUMMARY OF BACKGROUND DATA: e beam-sterilized PLDLLA cages were shown to maintain mechanical strength for at least 6 months during degradation studies in vitro. Yet failure of the cages was observed after only 3 months in vivo. We hypothesized that degradation characteristics and mechanical strength could be improved by sterilizing the cages through ethylene oxide (EtO) instead of e-beam. METHODS: PLDLLA cages were sterilized either by e-beam or EtO, and degraded in phosphate-buffered saline. Each month, cages were compressed until failure. Inherent viscosity was determined as a measure of degradation. For the in vivo evaluation, e-beam- or EtO-sterilized cages were implanted at L3-L4 in a standardized goat model. After 3 or 6 months, retrieved segments were scanned by high-resolution magnetic resonance imaging. Also, inherent viscosity of the polymer was measured. RESULTS: e-beam sterilization strongly decreased inherent viscosity of PLDLLA compared with EtO sterilization, but initial strength was only affected marginally. After 6 months, the strength of the e-beam-sterilized cages dropped, while that of EtO sterilized cages was maintained. Degradation in vivo was slightly faster than in vitro. In both groups, however, mechanical failure occurred at 3 months after implantation. CONCLUSIONS: Inherent viscosity decreases with degradation time, but strength only decreases when inherent viscosity is below a certain threshold. Above this threshold, mechanical strength is a property of the polymer and independent of inherent viscosity. e-beam sterilization strongly decreases inherent viscosity and thus advances mechanical degradation. EtO sterilization delays degradation but does not increase initial strength. Early failure of PLDLLA cages in the goat model thus is unrelated to sterilization method and requires further study. PMID- 17414908 TI - Intradiscal pressure, shear strain, and fiber strain in the intervertebral disc under combined loading. AB - STUDY DESIGN: Finite element study. OBJECTIVE: To investigate intradiscal pressure, shear strain between anulus and adjacent endplates, and fiber strain in the anulus under pure and combined moments. SUMMARY OF BACKGROUND DATA: Concerning anulus failures such as fissures and disc prolapses, the mechanical response of the intervertebral disc during combined load situations is still not well understood. METHODS: A 3-dimensional, nonlinear finite element model of a lumbar spinal segment L4-L5 was used. Pure unconstraint moments of 7.5 Nm in all anatomic planes with and without an axial preload of 500 N were applied to the upper vertebral body. The load direction was incrementally changed with an angle of 15 degrees between the 3 anatomic planes to realize not only moments in the principle motion planes but also moment combinations. RESULTS: Intradiscal pressure was highest in flexion and lowest in lateral bending. Load combinations did not increase the pressure. A combination of lateral bending plus flexion or lateral bending plus extension strongly increased the maximum shear strains. Lateral bending plus axial rotation yielded the highest increase in fiber strains, followed by axial rotation plus flexion or axial rotation plus extension. The highest shear and fiber strains were both located posterolaterally. An additional axial preload tended to increase the pressure, the shear, and fiber strains essentially for all load scenarios. CONCLUSIONS: Combined moments seem to lead to higher stresses in the disc, especially posterolaterally. This region might be more susceptible to disc failure and prolapses. These results may help clinicians better understand the mechanical causes of disc prolapses and may also be valuable in developing preventive clinical strategies and postoperative treatments. PMID- 17414909 TI - Musculotendon and fascicle strains in anterior and posterior neck muscles during whiplash injury. AB - STUDY DESIGN: A biomechanical neck model combined with subject-specific kinematic and electromyographic data were used to calculate neck muscle strains during whiplash. OBJECTIVES: To calculate the musculotendon and fascicle strains during whiplash and to compare these strains to published muscle injury thresholds. SUMMARY OF BACKGROUND DATA: Previous work has shown potentially injurious musculotendon strains in sternocleidomastoid (SCM) during whiplash, but neither the musculotendon strains in posterior cervical muscles nor the fascicle strains in either muscle group have been examined. METHODS: Experimental human subject data from rear-end automobile impacts were integrated with a biomechanical model of the neck musculoskeletal system. Subject-specific head kinematic data were imposed on the model, and neck musculotendon and fascicle strains and strain rates were computed. Electromyographic data from the sternocleidomastoid and the posterior cervical muscles were compared with strain data to determine which muscles were being eccentrically contracted. RESULTS: SCM experienced lengthening during the retraction phase of head/neck kinematics, whereas the posterior muscles (splenius capitis [SPL], semispinalis capitis [SEMI], and trapezius [TRAP]) lengthened during the rebound phase. Peak SCM fascicle lengthening strains averaged (+/-SD) 4% (+/-3%) for the subvolumes attached to the mastoid process and 7% (+/-5%) for the subvolume attached to the occiput. Posteriorly, peak fascicle strains were 21% (+/-14%) for SPL, 18% (+/-16%) for SEMI, and 5% (+/-4%) for TRAP, with SPL strains significantly greater than calculated in SCM or TRAP. Fascicle strains were, on average, 1.2 to 2.3 times greater than musculotendon strains. SCM and posterior muscle activity occurred during intervals of muscle fascicle lengthening. CONCLUSIONS: The cervical muscle strains induced during a rear-end impact exceed the previously-reported injury threshold for a single stretch of active muscle. Further, the larger strains experienced by extensor muscles are consistent with clinical reports of pain primarily in the posterior cervical region following rear-end impacts. PMID- 17414910 TI - A biomechanical assessment of thoracic spine stapling. AB - STUDY DESIGN: An in vitro immature bovine study of thoracic spine fixation using shape memory alloy for minimally invasive adolescent scoliosis treatment. OBJECTIVE: The purpose of this study was to detect differences in thoracic spine range of motion due to the placement of spinal staples manufactured from a common shape memory alloy. SUMMARY OF BACKGROUND DATA: Scoliosis surgery attempts to both correct the existing deformity and prevent further curve progression. The gold standard in surgical treatment of scoliosis is posterior instrumentation and fusion using pedicle screws or hooks. Fusionless techniques generally use less invasive procedures than fusion methods. One such technique, anterior stapling of the vertebrae, theoretically enables preservation of growth, motion, and spinal function. However, the degree of stability afforded by this method has not been reported. METHODS: Eight immature bovine specimens (T4-T9) were used in this study. Nitinol staples were cooled to below the transition temperature and placed on the spine such that they spanned the disc space. The specimen was then heated above the critical temperature and the staple tines engaged the vertebral endplates. A common flexibility protocol was used to determine the ranges of motion (ROM) in flexion-extension, lateral bending, and axial rotation. The intact spine and 4 construct variants (combinations of staple type and placement) were evaluated using this testing protocol. ANOVA statistics with post hoc testing was used to discern statistical differences. RESULTS: Not all staple variants were able to achieve significant reductions with respect to the intact condition. ROM was significantly restricted in axial rotation and lateral bending with the introduction of staple instrumentation. Further, there seemed to be a mechanical equivalence between a single double-prong staple and 2 single-prong staples. Our data indicate that staple fixation does not result in consistently elevated adjacent segment motion. CONCLUSIONS: Our results imply that staples are able to significantly restrict motion while not achieving motion reductions that one would achieve with fusion-promoting instrumentation. The choice between double- and single-prong staples remains a matter of preference. Neither staple variant provided a mechanical advantage. The single-prong staple did allow more control in the placement of the staple over the disc space. The addition of an anterior staple significantly reduced the overall flexion-extension ROM. PMID- 17414911 TI - Anterior cervical discectomy and fusion without instrumentation. AB - STUDY DESIGN: Review of clinical file information and postoperative imaging, collected prospectively over a period of 14 years, in anticipation of study. OBJECTIVES: 1) Assessment of technical success in achieving anterior cervical fusion without internal fixation; 2) assessment of postoperative neck pain relevant to technical success or failure of fusion; and 3) assessment of morbidity arising from iliac crest bone graft donor site. SUMMARY OF BACKGROUND DATA: After anterior cervical discectomy and bone grafting for cervical radiculopathy or the intractable pain of cervical spondylosis, common clinical practice varies widely between the extremes of internal fixation in all cases, and never applying fixation. The clinical information and relevant imaging of 97 consecutive patients, 46 male, was reviewed at 12 months after surgery. METHODS: All surgery was performed at no more than 2 contiguous levels, by one surgeon (S.M.E.). After anterior discectomy alone, or combined with posterior vertebral body margin osteophytectomy, anterior bone grafting (Smith-Robinson) was performed at each level using a tricortical autogenous iliac crest bone block inserted under compression. In the interests of maximizing resource allocation and minimizing potential complications, all surgery was completed without internal fixation. A postoperative semirigid cervical collar was prescribed for 2 months. RESULTS: In 54 patients having 1-level fusion, there were 6 pseudarthroses (11%). In 43 patients having 2-level fusion, 12 patients demonstrated pseudarthroses (28% of patients) at a total of 18 levels (21% of levels). Only 2 of the 97 patients had pain related to the donor site. CONCLUSIONS: These results tend to confirm published reports of high pseudarthrosis rates in anterior cervical fusions carried out at 2 or more levels without fixation, as against improved fusion rates when internal fixation is applied. The authors are inclined to change their practice to include internal fixation in the form of anterior plating for fusions carried out at more than one level. Patients with technically successful fusions were less likely to have postoperative neck pain. Donor site pain was not a significant postoperative complication. PMID- 17414913 TI - Incidence and outcome of whiplash injury after multiple trauma. AB - STUDY DESIGN: A retrospective study of 101 consecutive polytrauma patients with regard to whiplash injury. OBJECTIVES: To investigate the incidence and evaluate long-term outcome of whiplash injury following high-energy trauma. SUMMARY OF BACKGROUND DATA: Chronic whiplash injury has been widely reported in the literature, following low-energy trauma. Very few studies exist on whiplash injury following high-energy trauma. METHODS: A total of 101 consecutive polytrauma patients admitted to our Level I Trauma Center over a 2-year period, fulfilling the inclusion criteria (age >18 years, high-energy trauma [a fall from a height >2 m, road traffic accidents with speed >30 km/h], and Injury Severity Score >16), were assessed. Whiplash injury was defined according to Quebec Task Force guidelines. The study group (n = 13) included patients who developed whiplash injury symptoms and the control group (n = 88) those who did not. The Neck Disability Index was calculated as an outcome measure for patients complaining of whiplash injury symptoms. The mean follow-up was 17 months. The chi2 and Student t tests were used for the statistical analysis (SPSS 12.1; SPSS, Inc., Chicago, IL). RESULTS: Only 13 out of 101 patients (1 female/12 male) (13%) complained of whiplash injury. There was a significantly higher rate of neck pain at triage (P < 0.001) and higher combined mean of Abbreviated Injury Score of upper torso (P < 0.0001) in the study group, elucidating the cause of whiplash injury. The Neck Disability Index was <24 points, indicating only mild-to moderate disability in these patients. Whiplash injury incidence in this study (13%) was similar to the incidence of neck pain in the general population. CONCLUSIONS: The incidence of whiplash injury following polytrauma was found to be low in our study. There is no dose-response relation between magnitude of trauma severity and incidence of whiplash injury. PMID- 17414914 TI - Total lumbar disc replacement: different results for different levels. AB - STUDY DESIGN: Prospective study analyzing midterm clinical results of total lumbar disc replacement (TDR) with ProDisc II performed at different lumbar motion segments. OBJECTIVES: To assess the influence of the disc level and number of discs replaced following TDR on postoperative outcome. SUMMARY OF BACKGROUND DATA: Multisegmental disc replacement procedures belong to the so-called "off label" indications for disc replacement, which still lack evidence of noninferiority when compared with fusion procedures. Results from uncontrolled clinical trials regarding monosegmental versus multisegmental disc replacements are contradictory. METHODS: The influence of the level and the number of lumbar discs replaced on postoperative outcome was analyzed prospectively according to Visual Analogue Scale (VAS), Oswestry Disability Index (ODI), and numerous clinical parameters. Post-TDR pain patterns were analyzed with fluoroscopically guided spine infiltrations. RESULTS: A total of 99 patients from 3 treatment groups with a mean follow-up of 25.8 months (range, 12.1-57.5 months) achieved significant improvement of preoperative VAS and ODI levels (P < 0.05). Best results and highest patient satisfaction rates (90.9%) were achieved in patients with monosegmental TDR at L4-L5 (n = 22). Results deteriorated when monosegmental TDR was performed at the lumbosacral junction (n = 57) with a tendency toward statistical significance at 24-month follow-up (P = 0.07). Postoperative outcome was significantly inferior following bisegmental disc replacements at L4-L5 + L5 S1 (n = 20) with a considerably higher complication rate when compared with monosegmental TDR procedures. Fluoroscopically guided spine infiltrations confirmed that the incidence of postoperative pain from posterior joint structures was 9.1% (n = 2) for L4-L5 TDR, 28.1% (n = 16) following L5-S1, and 60.0% (n = 12) for bisegmental-TDR at L4-L5 + L5-S1, respectively. CONCLUSION: The level and the number of lumbar disc replacements influence postoperative outcome significantly. Satisfactory outcome was achieved for monosegmental L4-L5 and L5-S1 disc replacement procedures with best results achieved following TDR at L4-L5. For bisegmental TDR, complication rates are significantly higher and inferior postoperative results are to be expected. The incidence of postoperative pain originating from facet and/or iliosacral joints is currently underestimated and will require further investigation. PMID- 17414915 TI - The influence of fracture mechanism and morphology on the reliability and validity of two novel thoracolumbar injury classification systems. AB - STUDY DESIGN: The Thoracolumbar Injury Severity Score (TLISS) and the Thoracolumbar Injury Classification and Severity Score (TLICS) were prospectively evaluated. OBJECTIVES: To compare the reliability and validity of the TLISS and TLICS schemes to determine the importance of injury mechanism and morphology to the identification and treatment of thoracolumbar fractures. SUMMARY OF BACKGROUND DATA: Two novel algorithms have been developed for the categorization and management of thoracolumbar injuries: the TLISS system emphasizing injury mechanism and the TLICS scheme involving injury morphology. METHODS: The clinical and radiographic findings of 25 patients with thoracolumbar fractures were prospectively presented to 5 groups of surgeons with disparate levels of training and experience with spinal trauma. These injuries were consecutively scored, first using the TLISS and then 3 months later with the TLICS. The recommended treatments proposed by the 2 schemes were compared with the actual management of each patient. RESULTS: For both algorithms, the interrater kappa statistics of all subgroups (mechanism/morphology, status of the posterior ligaments, total score, predicted management) were within the range of moderate to substantial reproducibility (0.45-0.74), and there were no statistically significant differences noted between the respective kappa values. Interrater correlation was higher for the TLISS paradigm on mechanism/morphology, integrity of the posterior ligaments, and proposed management (P < or = 0.01). The TLISS and TLICS schemes both exhibited excellent overall validity. CONCLUSIONS: Although both schemes were noted to have substantial reproducibility and validity, our results indicate the TLISS is more reliable than the TLICS, suggesting that the mechanism of trauma may be a more valuable parameter than fracture morphology for the classification and treatment thoracolumbar injuries. Since these injury characteristics are interrelated and are critical to the maintenance of spinal stability, we think that both concepts should be considered during the assessment and management of these patients. PMID- 17414916 TI - The sensitivity of review results to methods used to appraise and incorporate trial quality into data synthesis. AB - STUDY DESIGN: Systematic review. OBJECTIVE: To determine whether results and conclusions on the effectiveness of exercise for workers with neck pain vary with the Cochrane Back Review Group Guidelines and best-evidence synthesis review methods. To identify methodologic weaknesses associated with these review methods that may impact on the validity of their results. SUMMARY OF BACKGROUND DATA: The Cochrane Back Review Group Guidelines and best-evidence synthesis have different approaches to appraising trial quality and incorporating quality into data synthesis. The impact of different review methods on the reproducibility and validity of review results is unknown. METHODS AND RESULTS: Systematic search of Medline, Embase, CINAHL, and Cochrane databases, without language restrictions. Twelve trials were selected. Two review methods were used to appraise trial quality and to incorporate quality into data synthesis. As recommended by the Cochrane Back Review Group Guidelines, trials were assigned quality scores using a scale. Results of all 12 trials were stratified into levels of evidence according to their scores. Based on these results, no treatment recommendation could be formulated. Best-evidence synthesis critically appraised methodology; trials were accepted on the strength of their scientific merit or rejected due to risk of bias. According to the 4 trials accepted for best-evidence synthesis, workers should be activated with exercise given its beneficial effect on patient perceived recovery. Both the Cochrane Back Review Group Guidelines and best evidence synthesis reviews were found to have weaknesses associated with their methods. CONCLUSIONS: Review results and conclusions are sensitive to methods for appraising trial quality and incorporating quality into data synthesis when the evidence consists largely of low-quality trials. Both the Cochrane Back Review Group Guidelines and best-evidence synthesis methods were found to have strengths and methodologic weaknesses that healthcare decision-makers should be aware of when interpreting systematic reviews. PMID- 17414917 TI - Key factors in back disability prevention: a consensus panel on their impact and modifiability. AB - STUDY DESIGN: Modified Delphi panel using Q-methodology. OBJECTIVE: To reach consensus on the relative impact and modifiability of factors that might prevent participation restrictions in people with back pain as defined by the WHO International Classification of Functioning, Disability and Health. SUMMARY OF BACKGROUND DATA: Back disability prevention should focus on factors with large impact that are relatively easy to change and supported by most stakeholders. An evidence-based consensus panel can provide interim direction until definitive evidence is available. METHODS: Evidence summaries for 32 factors were used by 33 researchers and stakeholders in a 3-round Delphi process to rank the factors' relative impact and modifiability. Consensus was judged as strong (>85% of panel members), moderate (50%-84%), or low (33%-49%). RESULTS: Most available research focused on return to work and often left unexplored other participation domains and environmental factors at home. The panel had substantial disagreements, particularly on the impact of changes to physical functioning and activities required at work. After 3 rounds, there was strong consensus that care provider reassurance had a high impact. There was moderate consensus that expectation of recovery and decreased fears had a high impact. Back supports, care provider reassurance, and patient knowledge were deemed most modifiable. CONCLUSIONS: Until definitive evidence is available, back disability prevention interventions will likely need to address multiple factors simultaneously and emphasize efforts to improve care provider reassurance. Consensus findings may vary depending on the panel. Research on nonoccupational participation, environmental factors at home, and reasons for discordant interpretation of evidence is needed. PMID- 17414918 TI - Systematic review of randomized trials comparing lumbar fusion surgery to nonoperative care for treatment of chronic back pain. AB - STUDY DESIGN: Systematic review of randomized trials comparing surgical to nonsurgical treatment of discogenic back pain. OBJECTIVE: Compare research methods and results. SUMMARY OF BACKGROUND DATA: Recent reports have increased debate about the role of surgery in the treatment of chronic back pain associated with lumbar disc degeneration. We conducted a systematic review of randomized trials comparing lumbar fusion surgery to nonsurgical treatment of chronic back pain associated with lumbar disc degeneration. METHODS: A literature search identified 5 randomized trials that compared fusion to nonoperative treatment for chronic low back pain. Excluding 1 trial for spondylolisthesis, we compared study participants, interventions, analyses, and outcomes in 4 trials that focused on nonspecific chronic back. RESULTS: All trials enrolled similar subjects. One study suggested greater improvement in back-specific disability for fusion compared to unstructured nonoperative care at 2 years, but the trial did not report data according to intent-to-treat principles. Three trials suggested no substantial difference in disability scores at 1-year and 2-years when fusion was compared to a 3-week cognitive-behavior treatment addressing fears about back injury. However, 2 of these trials were underpowered to identify clinically important differences. The third trial had high rates of cross-over (>20% for each treatment) and loss to follow-up (20%); it is unclear how these affected results. CONCLUSIONS: Surgery may be more efficacious than unstructured nonsurgical care for chronic back pain but may not be more efficacious than structured cognitive-behavior therapy. Methodological limitations of the randomized trials prevent firm conclusions. PMID- 17414919 TI - Bone morphogenetic proteins and bone defects: a systematic review. AB - STUDY DESIGN: A systematic review of scientific literature. OBJECTIVE: The study aimed to determine whether bone morphogenetic proteins (BMPs) are more effective in treating bone defects than traditional techniques, such as grafting autologous bone. SUMMARY OF BACKGROUND DATA: BMPs were used in several human randomized controlled trials (RCTs). There are both logical arguments and an empirical basis for using RCTs to evaluate the effects of health care interventions and restrict systematic reviews to such a kind of study design. METHODS: An electronic search was made in the databases of MEDLINE, EMBASE (through MeSH and Emtree), and the Cochrane Central Register of Controlled Trials, extended to May 31, 2006, with no linguistic restrictions. RCTs that compare bone regeneration achieved through BMPs versus that obtained by traditional methods entered the study. RESULTS: The 17 publications that met the criteria, divided into subgroups by type of bone, were tabulated by salient characteristics and evaluated through the items proposed by van Tulder et al. However, as the studies differed widely (in terms of site, sample size, dosage of active principle, carrier, clinical and radiologic data recording), it was possible to carry out a metaanalysis of clinical and radiologic outcome only for the subgroup that evaluated the vertebrae, where it was observed that BMPs offer a slightly but statistically significant greater efficacy than do traditional techniques. CONCLUSIONS: The use of BMPs at the vertebrae can eliminate the need for surgery to harvest autologous bone. The only large study carried out on the other sites suggests that BMPs should be used at a concentration of 1.5 mg/mL to treat fractures of the tibia. However, further RCTs of good methodological quality are advisable so as to clarify the effectiveness of BMPs in clinical practice. PMID- 17414920 TI - Re: Villarraga ML, Cripton PA, Teti SD, et al. Wear and corrosion in retrieved thoracolumbar posterior internal fixation. Spine 2006;31:2454-62. PMID- 17414921 TI - Association between sciatica and microbial infection: true infection or culture contamination? PMID- 17414924 TI - Re: Whitman JM, Flynn TW, Childs JD, et al. A comparison between two physical therapy treatment programs for patients with lumbar spinal stenosis: a randomized clinical trial. Spine 2006;31:2541-9. PMID- 17414926 TI - Interferon gamma-inducible protein 10: a predictive marker of successful treatment response in hepatitis C virus/HIV-coinfected patients. AB - BACKGROUND: Elevated pretreatment interferon (IFN) gamma-inducible protein 10 (IP 10/CXCL10) levels are a marker of treatment nonresponse in hepatitis C virus (HCV)-monoinfected patients. We undertook this study to determine if IP-10 is a marker of treatment outcome in HCV/HIV-coinfected patients. METHODS: Nineteen HCV/HIV-coinfected patients were treated with weight-based pegylated (PEG) IFNalpha-2b (1.5 microg/kg) once weekly plus weight-based ribavirin (1000 or 1200 mg) daily for up to 48 weeks. Plasma IP-10, monokine induced by IFNgamma/CXCL9 (Mig), and IFN-inducible T-cell alpha-chemoattractant/CXCL11 (I-TAC) levels were measured by enzyme-linked immunosorbent assay on samples obtained frequently during the first 3 PEG-IFN doses and throughout treatment. RESULTS: Median pretreatment plasma IP-10 (interquartile range [IQR]) levels were significantly lower in virological responders (n=6) at 217 (IQR: 181-301) pg/mL compared with nonresponders (n=13) at 900 (IQR: 628-2048) pg/mL (P=0.002), whereas pretreatment Mig and I-TAC levels did not differ significantly. Plasma IP-10 levels of 400 pg/mL before treatment and on days 7 and 14 could be used to identify likely coinfected PEG-IFN/ribavirin nonresponders. PEG-IFN-induced elevations in IP-10 were greater in virological responders than in nonresponders (approximately 10 fold vs. approximately 4-fold) after the first PEG-IFN dose. CONCLUSIONS: IP-10 may be a biomarker of HCV treatment outcome in difficult-to-treat HCV/HIV coinfected patients. PMID- 17414927 TI - Clinically significant anemia in HIV-infected pregnant women in India is not a major barrier to zidovudine use for prevention of maternal-to-child transmission. AB - OBJECTIVES: To determine the prevalence of anemia (serum hemoglobin <10 g/dL) and assess zidovudine use and toxicity in HIV-positive pregnant women in India. METHODS: From 2002 through 2006, 24,105 pregnant women in Pune were screened for HIV and anemia. As part of an infant prevention of mother-to-child transmission (PMTCT) trial, enrolled HIV-positive women (n = 467) were assessed for anemia and associated outcomes, comparing women receiving zidovudine for >or=2 weeks versus no zidovudine. RESULTS: The prevalence of anemia was 38.7% in HIV-positive women. Anemic women were as likely as nonanemic women to receive zidovudine. At delivery, regardless of anemia status at enrollment, women receiving >or=2 weeks of zidovudine were 70% less likely to be anemic compared with women receiving no zidovudine (odds ratio = 0.28, 95% confidence interval: 0.14 to 0.57; P < 0.01), received iron and folic acid supplements for longer periods, and had no increased adverse delivery or newborn birth outcomes. CONCLUSIONS: A significant proportion of HIV-positive pregnant women in India present for antenatal care with anemia. With concurrent iron and folic acid supplementation, however, zidovudine use is not associated with persistent or worsening anemia or associated adverse outcomes. In Indian community settings, all pregnant HIV-positive women should receive early anemia treatment. Mild anemia should not limit zidovudine use for PMTCT in India. PMID- 17414928 TI - The changing pattern of hospital care for persons living with HIV: 2000 through 2004. AB - OBJECTIVE: To compare inpatient utilization and costs by persons living with HIV in 2000 with inpatient utilization and costs in 2004. DATA SOURCES: Data on 91,343 hospital discharge abstracts representing all HIV-related admissions in 6 states (California, Florida, New Jersey, New York, South Carolina, and Washington state) in 2000 and data from 72,829 hospital discharge abstracts representing all HIV-related admissions in the same states in 2004 are used. These data were obtained from the Agency for Healthcare Research and Quality's Healthcare Cost and Utilization Project, and they were combined with data on the number of persons living with HIV that were obtained from the Centers for Disease Control and Prevention and 2 state departments of health. STUDY DESIGN: This study compares the hospital care received by persons living with HIV in 6 states in calendar year 2000 with the hospital care received by persons living with HIV in calendar year 2004 in the same 6 states. This study also compares population based measures of hospital utilization (ie, to measure the average utilization of hospital care per person living with HIV in each state) across the 6 states. RESULTS: This study found that the average age of a hospitalized patient with HIV rose from 41 to 44 years and that the average number of diagnoses rose from 6.0 to 7.4. Moreover, it was found that the average number of admissions per person living with HIV fell 39% and that the percentages of female and black patients with HIV remained the same. CONCLUSIONS: Hospitalized patients living with HIV are getting older and sicker, although the average number of admissions per person living with HIV continues to fall. PMID- 17414929 TI - Pharmacokinetics of emtricitabine, tenofovir, and GS-9137 following coadministration of emtricitabine/tenofovir disoproxil fumarate and ritonavir boosted GS-9137. AB - OBJECTIVES: To evaluate the potential for clinically relevant drug-drug interaction between emtricitabine/tenofovir disoproxil fumarate (FTC/TDF) and the ritonavir-boosted HIV integrase inhibitor GS-9137 (GS-9137/r). METHODS: Healthy adults were administered FTC/TDF (200/300 mg once daily) for 7 days, followed by randomization to the order of receiving GS-9137/r (50/100 mg once daily) and GS 9137/r plus FTC/TDF in a crossover fashion under fed conditions for 10 days. Pharmacokinetic (PK) blood draws were performed on days 7, 17, and 27. Lack of PK alteration for FTC, tenofovir (TFV), and GS-9137 was defined as a 90% confidence interval (CI) for the estimated ratio of geometric least squares means (coadministration/alone) between 70% and 143% for the primary PK parameters: maximum observed plasma concentration (Cmax), area under the plasma concentration time curve over dosing interval (AUCtau), and trough concentration (Ctau). RESULTS: Twenty-four of the 26 enrolled subjects completed the study with no serious adverse events or discontinuations attributable to adverse events. FTC, TFV, and GS-9137 PKs were unaffected during coadministration, with Cmax, AUCtau, and Ctau, meeting the protocol definition of equivalence and also the stricter bioequivalence criteria (90% CI: 80% to 125%). FTC and TFV PK parameters were comparable to historical values. CONCLUSION: There is no clinically relevant drug drug interaction between FTC/TDF and GS-9137/r on their coadministration. PMID- 17414930 TI - Effect of micronutrients and iron supplementation on hemoglobin, iron status, and plasma hepatitis C and HIV RNA levels in female injection drug users: a controlled clinical trial. AB - BACKGROUND: Iron deficiency is common among female injection drug users, but it is unclear whether iron supplementation can reduce anemia and improve iron status without increasing plasma hepatitis C virus (HCV) or HIV RNA levels. METHODS: We conducted a phase 3, double-blind, randomized, controlled clinical trial of daily micronutrients with 18 mg of iron (iron group) versus micronutrients without iron (control group) for 12 months among hepatitis C-positive female injection drug users in Baltimore, Maryland. The main outcome measures were hemoglobin, markers of iron status, plasma HCV RNA, plasma HIV RNA, and liver enzymes at 6 and 12 months of follow-up. RESULTS: Four hundred fifty-eight women (320 HIV-negative and 138 HIV-positive) enrolled in the trial. There were no significant differences in the proportion of women with anemia, ferritin<30 ng/mL, log10 plasma HCV RNA, or log10 plasma HIV RNA between treatment groups at enrollment. The proportion with anemia in the iron and control groups, respectively, was 20.7% versus 31.3% (P=0.026) at 6 months and 26.2% versus 30.4% (P=0.5) at 12 months; with ferritin<30 ng/mL, the proportion was 29.2% versus 55.5% (P<0.0001) at 6 months and 26.2% versus 46.9% (P=0.0018) at 12 months. In the iron and control groups, respectively, mean log10 plasma HCV RNA (IU/mL) was 5.2 versus 5.2 (P=0.86) at 6 months and 5.4 versus 5.3 (P=0.6) at 12 months. Among HIV positive subjects, mean log10 plasma RNA (copies/mL) in the iron and placebo groups, respectively, was 3.8 versus 3.7 (P=0.75) at 6 months and 3.7 versus 4.1 (P=0.19) at 12 months. There were no significant differences in liver enzyme levels between the treatment groups at enrollment, 6 months, and 12 months. CONCLUSIONS: A daily micronutrient supplement with iron can reduce anemia and improve iron status in female injection drug users without increasing plasma HCV or HIV RNA levels or altering liver enzymes. PMID- 17414931 TI - Antiretroviral durability and tolerability in HIV-infected adults living in urban Kenya. AB - BACKGROUND: Insufficient data exist on the durability and tolerability of first line antiretroviral therapy (ART) regimens provided by HIV treatment programs implemented in developing countries. METHODS: Longitudinal observation of clinical, immunologic, and treatment parameters of all HIV-infected adult patients initiated on ART was performed at Saint Mary's Mission Hospital in Nairobi, Kenya from September 2004 until August 2006. RESULTS: A total of 1286 patients were analyzed (59.1% female). Initial ART regimens were primarily stavudine, lamivudine, and nevirapine (62.1%). Median ART duration was 350 days (11.6 months). Significant improvements in clinical and immunologic status were noted after 12 months of therapy. ART switches occurred in 701 (54.5%) patients. The cumulative incidence of ART switch at 12 months was 78.4%. Concurrent ART related toxicities (40.6%) and tuberculosis treatment interactions (28.1%) were the most frequent reasons for ART switch. Baseline AIDS symptoms (hazard rate [HR]=1.59, 95% confidence interval [CI]: 1.28 to 1.98; P<0.01) and a CD4 countor=800 CD4 cells/mm in 5299 therapy-naive patients starting HAART, (2) CD4 cell count changes during 7 years of uninterrupted HAART in a subset of 544 patients, and (3) reaching a plateau in CD4 cell restoration after 5 years of HAART in 366 virologically suppressed patients. RESULTS: Among patients with <50, 50 to 200, 200 to 350, 350 to 500, and >or=500 CD4 cells/mm at baseline, respectively, 20%, 26%, 46%, 73%, and 87% reached >or=800 CD4 cells/mm within 7 years of starting HAART. Periods with HIV RNA levels >500 copies/mL and age >or=50 years were associated with lesser increases in CD4 cell counts between 6 months and 7 years. Having reached >or=800 CD4 cells/mm at 5 years, age >or=50 years, and >or=1 HIV RNA measurement >1000 copies/mL between 5 and 7 years were associated with a plateau in CD4 cell restoration. CONCLUSIONS: Restoration to CD4 cell counts >or=800 cells/mm is feasible within 7 years of HAART in most HIV-infected patients starting with >or=350 cells/mm and achieving sufficient suppression of viral replication. Particularly in patients >or=50 years of age, it may be beneficial to start earlier than current guidelines recommend. PMID- 17414937 TI - Genotyping of Chlamydia trachomatis from endocervical specimens in Brazil. AB - BACKGROUND: There is no data concerning genotyping of Chlamydia trachomatis from Brazilian samples. GOAL: To characterize the genotype of C. trachomatis detected in women assisted at a STD public clinic and establish the prevalence of this infection in that population. STUDY DESIGN: Endocervical samples of a group of 100 women were tested for chlamydial infection with PCR directed to C. trachomatis cryptic plasmid. Genotyping of positive samples were done after omp1 amplification and sequencing. RESULTS: The overall prevalence of C. trachomatis infection was 19%, with the highest prevalence in women between 15 and 25 years old (68.4%). Four genotypes were found associated with endocervical infections: D, E, F, and K. Sequence analysis revealed a coinfection of genotypes D and E in 1 woman. CONCLUSIONS: To our knowledge this is the first study to characterize Brazilian C. trachomatis endocervical samples and Brazilian C. trachomatis genotype coinfection. Our results also emphasize the importance of routine diagnosis of C. trachomatis for the control of this STD. PMID- 17414938 TI - Effects of pharmacological therapy on gait and cognitive function in depressed patients. AB - OBJECTIVE: To investigate the relationship among affective status, cognitive function, and gait in depressed patients and to evaluate the effects of treatment of depression on gait and cognitive function. METHODS: Nineteen patients recently diagnosed with clinical depression (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria) were recruited from a psychiatric outpatient clinic. Evaluation included the Hamilton Depression Rating Scale (HAM-D), the Mini-Mental State Examination, a computerized neuropsychological battery (Mindstreams, NeuroTrax Corp, New York, NY), and Barthel's Index of Instrumental Activities of Daily Living. Temporal parameters of gait were quantified using a stopwatch and force-sensitive insoles. All assessments were performed at baseline and after approximately 10 weeks of treatment with antidepressants. RESULTS: The patients' mean age was 68.6 +/- 9.1 years (15 women). Therapy significantly (P < 0.001) improved the affective state (HAM-D scores). There were small but significant improvements in gait speed (P = 0.033), stride time variability (P = 0.036), and gait asymmetry (P = 0.038). With the exception of the hand-eye coordination index, all tested cognitive domains also improved significantly. Baseline depression scores correlated with changes in depression: patients with higher HAM-D scores at baseline had more significant improvement in their affect (P < 0.001). Changes in HAM-D were not significantly correlated with changes in gait or changes on computerized tests of cognitive function (P > 0.10). CONCLUSIONS: Depressive symptoms are associated with gait and cognitive impairment. Moreover, the present results suggest that these domains improve in response to antidepressant medication. PMID- 17414939 TI - Pramipexole in levodopa-treated Parkinson disease patients of African, Asian, and Hispanic heritage. AB - BACKGROUND: Little is known regarding the effects of antiparkinsonian drugs in US racial or ethnic minorities. OBJECTIVE: : To evaluate the safety, tolerability, and efficacy of adjunctive pramipexole in Parkinson disease (PD) patients of African, Asian, or Hispanic heritage stably treated with levodopa. DESIGN: Multicenter, parallel-group, double-blind, randomized, placebo-controlled trial. SETTING: : Seventeen Parkinson Study Group sites in the United States and Puerto Rico. PATIENTS: One hundred forty-four PD patients of African, Asian, or Hispanic heritage enrolled from January 1997 to August 1998 and observed until October 1998. INTERVENTION: Subjects received pramipexole or placebo (3:1 ratio), 0.375 mg/d to a maximum tolerated dose ( or = 34) in RT it is important to consider the possibility of a dual HIV-1 infection. PMID- 17415036 TI - Intensification of a triple-nucleoside regimen with tenofovir or efavirenz in HIV 1-infected patients with virological suppression. AB - OBJECTIVE: To compare a quadruple-nucleoside with an efavirenz-containing regimen for treatment of HIV-1 infection. DESIGN: A randomized, open-label study of the AIDS Clinical Trials Group (ACTG). METHODS: Subjects receiving zidovudine/lamivudine/abacavir on ACTG 5095 with HIV-1 RNA less than 200 copies/ml were randomly assigned to intensify either with tenofovir or efavirenz. Subjects were followed for time to treatment failure, defined as either virological failure or treatment discontinuation. Analyses were intent-to-treat. RESULTS: One hundred and seventy subjects (21% women; 56% non-white) entered the study. At baseline, 95 and 73% had HIV-1-RNA levels less than 200 and 50 copies/ml, respectively; the median CD4 cell count was 453 cells/microl. Over a median 79 weeks follow-up, 165 (97%) completed the study, three (2%) discontinued, and two (1%) died. Treatment failure occurred in 31 subjects: 18 (21%) (quadruple nucleosides) and 13 (15%) (efavirenz-containing regimen); however the failure-time curves crossed and demonstrated a non-constant treatment effect over time, characterized by more early treatment failures on the efavirenz containing regimen and more late treatment failures on the four-nucleoside regimen. HIV-1 RNA remained suppressed in more than 88% of subjects to less than 200 copies/ml and in more than 78% to less than 50 copies/ml at weeks 24, 48, and 72, without differences by treatment arm. There were no significant differences between the regimens in CD4 cell increases, time to new grade 3/4 adverse events, or adherence. CONCLUSION: The safety, tolerability, and efficacy of the four nucleoside regimen were not significantly different from the efavirenz-containing regimen. These pilot data support further investigation of the quadruple nucleoside regimen. PMID- 17415037 TI - Triple-class antiretroviral drug resistance: risk and predictors among HIV-1 infected patients. AB - BACKGROUND: HIV-1 triple-class antiretroviral drug resistance (TC-DR) may substantially limit therapeutic options and compromise clinical outcomes. OBJECTIVE: To estimate TC-DR prevalence and incidence, and identify risk factors for TC-DR acquisition. METHODS: We identified patients in the University of North Carolina HIV Cohort Study with TC-DR HIV-1 variants. Nucleos(t)ide reverse transcriptase inhibitor (NRTI), non-nucleoside reverse transcriptase inhibitor (NNRTI), and major protease inhibitor (PI) mutations, were based on the International AIDS Society - USA guidelines. Prevalence was estimated with the exact binomial distribution, incidence with the exact Poisson distribution, and multivariable analyses were performed using logistic regression. RESULTS: Of 1587 patients, half initiated therapy with HAART (N = 789), 20% (N = 320) with non HAART combination therapy, and 30% (N = 478) with one NRTI. The median time on therapy was 5.7 years [interquartile range (IQR) 2.9, 8.6], the median number of previous antiretroviral agents was six (IQR 4, 8), and 47% (N = 752) were exposed to at least one NRTI, NNRTI and PI. Assuming patients without genotypes did not harbor TC-DR virus, the prevalence of TC-DR among all antiretroviral-experienced patients was 8% [95% confidence interval (CI) 6%, 9%]. The prevalence was 3% (95% CI 2%, 4%) and 12% (95% CI 10%, 15%) among patients treated initially with HAART and non-HAART, respectively. The number of antiretroviral agents received and initiating therapy with non-HAART or an unboosted PI, increased TC-DR risk in multivariable analyses. CONCLUSION: The majority of patients with TC-DR have extensive antiretroviral exposure, particularly to non-HAART regimens, whereas HAART initiators are at low risk of acquiring TC-DR during a median of 4 years of follow-up. PMID- 17415038 TI - Sex differences in the clinical, immunological and virological parameters of HIV infected patients treated with HAART. AB - OBJECTIVE: To compare the clinical, virological and immunological parameters of men and women at baseline and during antiretroviral treatment. METHODS: Analysis over time of data collected prospectively from of 2620 patients in a large cohort of HIV-infected patients followed for 12 months after initiating a nelfinavir based antiretroviral regimen. RESULTS: Women had higher CD4 cell counts (P < 0.001), lower viral load (P < 0.001) and more favourable clinical profile (P < 0.001) than men at baseline. Following treatment, antiretroviral drug-naive women had higher CD4 cell count (P = 0.01) over time than drug-naive men but similar virological responses (P = 0.6); among drug-experienced individuals, women had also better immunological (P = 0.06) and similar virological (P = 0.3) responses compared with men. Consequently, the viroimmunological profile was significantly more favourable in women at each time point. The rates of clinical progression or death were also lower in women (P = 0.008), although drug toxicity was observed more commonly in women (P = 0.09). The highest viroimmunological responses were observed during the first 3 months of therapy in both sexes, although virological responses were achieved up to the 6th month in drug-naive patients. Sex was significantly associated with clinical (P = 0.01), virological (P = 0.01) and immunological (P = 0.006) responses to antiretroviral treatment in multivariate analyses after adjustment for other variables. The differences between genders were not explained by different adherence to therapy. CONCLUSIONS: Women have more favourable clinical and viroimmunological patterns than men both at baseline and during antiretroviral treatment. Sex has a small but significant influence on the clinical and laboratory outcomes of HIV infection. PMID- 17415039 TI - The impact of male circumcision on HIV incidence and cost per infection prevented: a stochastic simulation model from Rakai, Uganda. AB - OBJECTIVES: To estimate the impact of male circumcision on HIV incidence, the number of procedures per HIV infection averted, and costs per infection averted. METHODS: A stochastic simulation model with empirically derived parameters from a cohort in Rakai, Uganda was used to estimate HIV incidence, assuming that male circumcision reduced the risks of HIV acquisition with rate ratios (RR) ranging from 0.3 to 0.6 in men, their female partners, and in both sexes combined, with circumcision coverage 0-100%. The reproductive number (R0) was also estimated. The number of HIV infections averted per circumcision was estimated from the incident cases in the absence of surgery minus the projected number of incident cases over 10 years following circumcision. The cost per procedure ($69.00) was used to estimate the cost per HIV infection averted. RESULTS: Baseline HIV incidence was 1.2/100 person-years. Male circumcision could markedly reduce HIV incidence in this population, particularly if there was preventative efficacy in both sexes. Under many scenarios, with RR < or = 0.5, circumcision could reduce R0 to < 1.0 and potentially abort the epidemic. The number of surgeries per infection averted over 10 years was 19-58, and the costs per infection averted was $1269-3911, depending on the efficacy of circumcision for either or both sexes, assuming 75% service coverage. However, behavioral disinhibition could offset any benefits of circumcision. CONCLUSION: Male circumcision could have substantial impact on the HIV epidemic and provide a cost-effective prevention strategy if benefits are not countered by behavioral disinhibition. PMID- 17415040 TI - Voluntary counselling and testing: uptake, impact on sexual behaviour, and HIV incidence in a rural Zimbabwean cohort. AB - OBJECTIVES: To examine the determinants of uptake of voluntary counselling and testing (VCT) services, to assess changes in sexual risk behaviour following VCT, and to compare HIV incidence amongst testers and non-testers. METHODS: Prospective population-based cohort study of adult men and women in the Manicaland province of eastern Zimbabwe. Demographic, socioeconomic, sexual behaviour and VCT utilization data were collected at baseline (1998-2000) and follow-up (3 years later). HIV status was determined by HIV-1 antibody detection. In addition to services provided by the government and non-governmental organizations, a mobile VCT clinic was available at study sites. RESULTS: Lifetime uptake of VCT increased from under 6% to 11% at follow-up. Age, increasing education and knowledge of HIV were associated with VCT uptake. Women who took a test were more likely to be HIV positive and to have greater HIV knowledge and fewer total lifetime partners. After controlling for demographic characteristics, sexual behaviour was not independently associated with VCT uptake. Women who tested positive reported increased consistent condom use in their regular partnerships. However, individuals who tested negative were more likely to adopt more risky behaviours in terms of numbers of partnerships in the last month, the last year and in concurrent partnerships. HIV incidence during follow-up did not differ between testers and non-testers. CONCLUSION: Motivation for VCT uptake was driven by knowledge and education rather than sexual risk. Increased sexual risk following receipt of a negative result may be a serious unintended consequence of VCT. It should be minimized with appropriate pre- and post-test counselling. PMID- 17415041 TI - Highly efficient HIV transmission to young women in South Africa. AB - BACKGROUND: Young women in sub-Saharan Africa are at very high risk of HIV acquisition, and high prevalence levels have been observed among women reporting one lifetime partner and few sexual contacts. Such findings have led to hypotheses that the probability of HIV transmission from men to women must be far higher than previously appreciated. METHODS: We used the data from a cross sectional national household survey of HIV among South African women aged 15-24 years to estimate the per-partnership transmission probability from men to women. Estimates were obtained using maximum likelihood methods and a transmission probability model allowing for random error in the self-reported number of lifetime partners. Sensitivity analyses were employed to assess the robustness of the per-partnership transmission probability estimates to the assumed HIV prevalence in male partners. RESULTS: HIV prevalence in women was 21.2% (95% confidence interval 17.9-24.5). The mean reported number of lifetime partners was 2.3. A significant increase in prevalence was observed with increasing lifetime partner numbers (P = 0.02). For a range of plausible values of the partner prevalence, the estimated per-partnership transmission probability varied from 0.74 to 1.00 with 95% confidence intervals ranging from 0.56 to 1.00. DISCUSSION: The per-partnership probability of HIV transmission from men to women in this population was very high. Before this, the majority of studies examining per partnership transmission probabilities estimated values below 50%. Identifying the factors that may drive the efficient spread of HIV in sub-Saharan Africa is essential for the development of effective prevention interventions. PMID- 17415042 TI - Monitoring death and cancer in children born to HIV-infected women in England and Wales: use of HIV surveillance and national routine data. AB - There may be long-term adverse health effects of in-utero antiretroviral therapy exposure. Data on children reported through national HIV surveillance were linked to routinely collected cancer and death data: a process known as "flagging". Ninety-five per cent (2612) of reported children born in 2001-2004 in England or Wales who were uninfected or of indeterminate infection status were flagged. By the end of 2005, no cancers and 14 deaths (three uninfected and 11 indeterminate) had been notified. PMID- 17415043 TI - Elevated plasma levels of high mobility group box protein 1 in patients with HIV 1 infection. AB - High mobility group box protein 1 (HMGB1) is a potent proinflammatory mediator. It has a dichotomic effect on HIV-1 replication in vitro but its role in vivo is unknown. Here we report the novel finding that plasma HMGB1 levels are elevated in HIV-1-infected patients, with the highest concentrations in patients with clinical complications. HMGB1 is likely to contribute to immunoactivation in HIV 1 infection in vivo. PMID- 17415044 TI - Primary genotypic resistance of HIV-1 to the maturation inhibitor PA-457 in protease inhibitor-experienced patients. AB - Sequences from 82 protease inhibitors (PI)-experienced patients were analysed for the presence of previously described in-vitro resistance mutations to PA-457 located in the C-terminal capside (H226Y, L231F, L231M) and in the N-terminal SP1 (A1V, A3T, A3V) within the CA-SP1 boundary domain. Overall, the CA-SP1 cleavage site was highly conserved in PI pre-treated patients and only one patient showed an L231M mutation. The impact of this mutation should be further addressed in vivo. PMID- 17415045 TI - Mortality after myocardial infarction in HIV-infected patients who have initiated HAART. AB - We identified all Danish HIV patients registered with myocardial infarction (MI) when on HAART (44 patients) and compared their mortality with that of matched patients with MI and no HIV and patients with HIV and no MI. Mortality in HIV infected MI patients was not significantly different to that of MI only patients in the first 90 days post-MI and thereafter was not significantly different to that of HIV patients without MI. PMID- 17415046 TI - Tuberculosis manifested by immune reconstitution inflammatory syndrome during HAART. AB - The proportion of tuberculosis manifested by immune reconstitution inflammatory syndrome (IRIS) after HAART has not been established. We describe the incidence and clinical features of tuberculosis manifested by IRIS after HAART in an intermediate tuberculosis burden area. The findings suggest that a significant proportion of the tuberculosis occurring early after starting HAART is manifested by IRIS. PMID- 17415047 TI - Treatment resistance after sequential interruption of a non-nucleoside reverse transcriptase inhibitor-based regimen. PMID- 17415048 TI - Effectiveness and durability of non-nucleoside reverse transcriptase inhibitor based therapy in HIV clinical practice. PMID- 17415049 TI - Immune reconstitution syndrome in tuberculosis and HIV-co-infected patients: Th1 explosion or cytokine storm? PMID- 17415050 TI - Atazanavir in plasma-exchange treatment. PMID- 17415051 TI - Partial splenic embolization as a treatment for hypersplenism in HIV/hepatitis C virus-co-infected patients. PMID- 17415052 TI - Child sexual abuse, HIV, and tuberculosis in sub-Saharan Africa: a case report from rural Cameroon. PMID- 17415053 TI - Response to Morrison et al., "Hormonal contraception and the risk of HIV acquisition". PMID- 17415054 TI - Loss of HIV-1-specific T-cell responses associated with very rapid HIV-1 disease progression. PMID- 17415055 TI - Polymorphisms in the 3' untranslated region of the fractalkine (CX3CL1) gene and the risk of HIV-1 infection and disease progression. PMID- 17415056 TI - Mycobacterium sherrisii: a new opportunistic agent in HIV infection? PMID- 17415057 TI - Health policy and practice management issues in orthopaedic surgery: editorial comment. PMID- 17415058 TI - Should operations be regionalized? The empirical relation between surgical volume and mortality. 1979. PMID- 17415060 TI - Disability evaluation: principles of treatment of compensable injuries. 1936. PMID- 17415061 TI - An epidemic of infantile paralysis. 1890. PMID- 17415062 TI - Orthopaedic procedure volume and patient outcomes: a systematic literature review. AB - The association between greater hospital procedure volumes and improved patient outcomes has been well established with respect to a variety of procedures and treatments. However, this association in orthopaedics has not been summarized systematically. We reviewed existing literature on associations between hospital and surgeon procedure volume and patient outcomes in orthopaedic surgery. The patient outcomes examined were mortality, hip dislocation, infection, revision, complications, functional outcome, and satisfaction. Of the 26 articles reviewed, most examined outcomes after primary joint arthroplasties (predominantly hip arthroplasties) with a relatively limited number of studies examining revision arthroplasties, hip fractures, spine, or general orthopaedics. No studies evaluated any other subspecialties. We found an association between higher hospital volumes and lower rates of mortality and hip dislocation. We also found an association between higher surgeon volume and lower rates of hip dislocation. All other associations were negative or inconclusive. In addition, surgeon volume had a greater effect on patients than hospital volume for primary and revision joint arthroplasties, whereas hospital volume was more strongly related to outcome than surgeon volume for the other procedures examined. Our findings suggest the need for additional studies in the various subspecialties to establish more definitive conclusions. PMID- 17415063 TI - Electronic medical records for the orthopaedic practice. AB - Electronic medical records (EMR) will be the standard method of medical documentation within the next decade. Making the conversion from paper to digital records will be costly in terms of actual cash outlay for an EMR system and the time investment in staff training, conversion of existing records and loss of operational efficiency during the implementation phase. The most difficult part of converting to EMR is ensuring the system you select will meet your practice's current objectives and those in years to come as technology--and the general population's adoption of it--changes. There are tremendous opportunities for current and future technologies to improve efficiency and reduce practice overhead, but selecting a system with limitations may make it difficult to realize these potentials. My aims are to: (1) explain why an EMR will be important to an orthopaedic practice and establish goals of EMR conversion; (2) outline attributes desirable in an EMR system; and (3) provide some insight regarding future direction of EMR. PMID- 17415064 TI - 50 years ago in CORR: Slipping of the upper femoral epiphysis. PMID- 17415066 TI - Is the diagnosis of melancholia important in shaping clinical management? PMID- 17415067 TI - Psychiatry benefits from psychopathology and diagnosis in shaping clinical management. PMID- 17415068 TI - The validity of melancholia. PMID- 17415069 TI - Melancholia: the importance of validity. PMID- 17415070 TI - Is the diagnosis of melancholia important in shaping clinical management? PMID- 17415071 TI - The development of the diagnostic and statistical manual of mental disorders version V substance use disorders section: establishing the research framework. PMID- 17415072 TI - Alcohol use among college students: an international perspective. AB - PURPOSE OF REVIEW: The present review of published articles during 2005-2006 on alcohol use among college students in Africa, Asia, Australasia, Europe and South America assesses the prevalence of alcohol use, hazardous drinking and related problems, and reviews the effectiveness of intervention methods and implications for future research. RECENT FINDINGS: Research on alcohol use and related problems in college students is lacking in many regions of the world. We identified 26 papers in peer-reviewed journals, from Australia, Brazil, Ecuador, Egypt, Germany, Hong Kong, Ireland, Lebanon, New Zealand, Nigeria, Sweden, The Netherlands and Turkey. SUMMARY: More comprehensive studies with systematic methodologies in the world regions reviewed here are needed to yield representative results on alcohol use and related risk and protective factors in college settings. College students in many countries are at elevated risk for heavy drinking, with serious immediate health risks, such as drink-driving and other substance use; and longer term risks, such as alcohol dependence. The prevalence of hazardous drinking in Australasia, Europe and South America appears similar to that in North America, but is lower in Africa and Asia. Alcohol policies should be reviewed and prevention programmes initiated in light of research evidence, for this high-risk population. PMID- 17415073 TI - The treatment of alcohol dependence. AB - PURPOSE OF REVIEW: The present review summarizes current research on the management of alcohol dependence, including pharmacotherapy, psychosocial interventions and treatment of alcohol dependence with comorbid psychiatric disorders. RECENT FINDINGS: Among recent studies, naltrexone has demonstrated the most consistent effect in reducing alcohol consumption in the context of behavioral therapy. In contrast to most previous studies, acamprosate did not show significant benefits on treatment outcomes relative to placebo. The combined use of naltrexone and acamprosate appeared to be safe and well tolerated but there was no additional therapeutic benefit. With the exception of topiramate, there are currently no new, effective medications for alcohol dependence. Of the psychosocial interventions, such as social behavior and network therapy, cognitive behavioral therapy, and motivational enhancement therapy, no one appears to be superior to another. Psychiatric comorbidity is common in alcohol dependent patients; however, there are too few studies to effectively guide treatment practice. SUMMARY: Progress has been made with pharmacotherapy and psychosocial interventions for alcohol-dependent individuals. More research is needed, however, in developing newer medications and psychosocial interventions in alcohol-dependent populations and in those with comorbid psychiatric conditions, and to improve the strategies to engage patients in continuing care. PMID- 17415074 TI - Cannabis use and psychiatric and cogitive disorders: the chicken or the egg? AB - PURPOSE OF REVIEW: Cannabis is the world's most commonly used illicit drug. In this review, we consider the recent literature on the effects of cannabis on mental health and on cognition. RECENT FINDINGS: Cannabis use in adolescence increases the risk of later schizophrenia-like psychoses, especially in genetically vulnerable individuals. Not surprisingly, patients already suffering from psychosis who use cannabis have a worse outcome than those who do not. These effects of cannabis may be consequent on its impact on the dopamine system. There is less evidence of cannabis playing an aetiological role in other mental disorders including depression, but there have been far fewer studies. Heavy cannabis use has also been shown to affect memory and learning performance, both in healthy individuals and in patients suffering from psychosis. Combined cognitive-behavioural therapy and motivational interviewing seems a promising psychological intervention to achieve a cessation of cannabis use in patients suffering from schizophrenia. SUMMARY: Further research is needed to understand the biological mechanisms underlying the effects of cannabis on mental health, but intervention strategies to help patients abstain should currently be implemented in psychiatric services, and public education campaigns should be directed at increasing awareness of the health risks of cannabis. PMID- 17415075 TI - Responding to the threat of HIV among persons with mental illness and substance abuse. AB - PURPOSE OF REVIEW: This article discusses current knowledge regarding the threat of HIV among persons with mental illness and substance abuse, and strategies for reducing this threat. It contains a review of the prevalence and consequences of dual/triple diagnosis, HIV risk behaviour and current HIV risk-reduction interventions among persons with dual diagnosis and interventions for triply diagnosed individuals. RECENT FINDINGS: Many persons with dual diagnosis remain undetected and there is a high prevalence of sexual risk behaviours among persons with dual diagnosis. Case management and supportive housing programmes are feasible options for the delivery of HIV risk-reduction interventions among such patients, and the adaptation of integrated behavioural treatment interventions can improve behavioural and healthcare utilization outcomes. SUMMARY: The developing world continues to see an escalation in HIV incidence. A more complete understanding of mental health, substance use and HIV serostatus interactions is needed to serve vulnerable populations. Mental health status not only mediates HIV risk behaviours, but positive serostatus has various effects on mental health. Co-morbid substance abuse is common among HIV-positive individuals with mental illness, resulting in serious adverse effects. Separate services for individuals with co-occurring substance abuse are less effective than integrated treatment programmes. PMID- 17415076 TI - Juvenile drug courts: emerging outcomes and key research issues. AB - PURPOSE OF REVIEW: In consideration of the widespread adoption of juvenile drug court programs during the past decade, the purpose of this review is to examine the effectiveness of juvenile drug courts and suggest priorities for juvenile drug court research. RECENT FINDINGS: Consistent with the much more extensive adult drug court literature and the few uncontrolled evaluations of juvenile drug court, findings from a recent randomized clinical trial suggest that juvenile drug court is more effective than family court in decreasing participant criminal behavior and substance use. Perhaps due to the intensive surveillance that juvenile drug court participants receive, however, these favorable outcomes did not translate to reduced rates of rearrest or incarceration during the 12-month study period. In addition, the integration of evidence-based substance-abuse treatments into juvenile drug court enhanced participant substance-related outcomes and rates of juvenile drug court completion. SUMMARY: Although the widespread dissemination of juvenile drug courts has exceeded clear and unambiguous evidence of their effectiveness, few other criminal justice programs have shown such promise with drug-abusing offenders. Moreover, the integration of evidence-based treatments of adolescent substance abuse holds the potential to further enhance the effectiveness of juvenile drug courts. PMID- 17415078 TI - Brain stimulation for the treatment of psychiatric disorders. AB - PURPOSE OF REVIEW: There has been a resurgence of interest in brain stimulation techniques as therapies for psychiatric disorders. Various names are used for this class of treatments: neuromodulation, somatic therapies, brain stimulation techniques. The methods in this class range from non-invasive (transcranial magnetic stimulation) to invasive brain surgery (deep brain stimulation). RECENT FINDINGS: Within the past year, the results of several large multicenter trials have been published, clearing the way for US Food and Drug Administration approval of vagus nerve stimulation for recurrent treatment-resistant depression and a pending consideration of approving transcranial magnetic stimulation for the treatment of depression. SUMMARY: This article reviews the most important recent clinically relevant manuscripts in this rapidly expanding new field. PMID- 17415079 TI - The neuropsychiatry of impulsivity. AB - PURPOSE OF REVIEW: Impulsive symptoms occur across neuropsychiatric disorders, with important ramifications for everyday functioning and quality of life. This article considers recent developments in the neuropsychological assessment of impulsivity with a focus on the ability to suppress motor responses (response inhibition). RECENT FINDINGS: Using objective tests, response inhibition deficits were identified in several neuropsychiatric conditions associated with impulsivity, namely attention deficit hyperactivity disorder, trichotillomania, obsessive-compulsive disorder, and chronic substance abuse. Deficits were also found in unaffected first-degree relatives of attention deficit hyperactivity disorder and obsessive-compulsive disorder patients. Evidence from patients with focal brain lesions and from healthy volunteers using functional MRI and transcranial stimulation implicated the right inferior frontal gyrus in response inhibition. Pharmacological manipulations of the serotonin system had no detectable behavioural effects on response inhibition, whereas manipulations of the noradrenaline system did. SUMMARY: Neuropsychological assessment shows great promise in the investigation of impulsivity and its brain substrates. These results support a key role for response inhibition, a function linked to the right inferior frontal gyrus, in the manifestation of impulsivity. Measures of response inhibition will contribute to the search for psychiatric endophenotypes, novel treatments, and more optimal diagnostic classification systems for neuropsychiatric disorders. PMID- 17415080 TI - Acurate diagnosis of delirium in elderly patients. AB - PURPOSE OF REVIEW: Delirium remains one of the most common complicating diagnoses in ailing elderly patients and a leading cause of morbidity, decreased quality of life, prolonged hospital stay, institutionalization and mortality. Despite its clinical importance and health-related costs, it often remains unrecognized or misdiagnosed. We evaluate currently available tools for the screening and diagnosis of delirium, their relevance and suitability for use in various clinical settings, as well as interobserver consistency amongst doctors and other nonclinician interviewers. RECENT FINDINGS: Extensive clinical trial evidence has been published recently concerning advances on the three fundamental elements of delirium assessment in elderly people: identification, severity assessment and reporting of existing predisposing and precipitating factors. SUMMARY: Despite advances on the pathophysiology and recognition of delirium, its detection relies on individual clinical expertise, a high index of suspicion and repeated cognitive testing of high-risk patients. Delirium diagnosis remains a clearly underresearched area; particularly, more work is required to adapt cognitive screening tools for use by nonclinicians, to develop cost-effective biochemical and molecular diagnostic techniques and to assess the effects of divulging updated consensus guidelines. PMID- 17415081 TI - Cognitive heterogeneity in schizophrenia. AB - PURPOSE: Further clarification of the nature of cognitive heterogeneity in schizophrenia is needed to aid the endophenotype approach to the understanding of the genetic basis of the disorder. This review summarizes recent neuropsychological studies of schizophrenia, aimed at establishing whether there are valid forms of cognitive impairment that can be defined with the use of neuropsychological measures in patients with schizophrenia, and studies that have attempted to relate specific neuropsychological findings to genetic polymorphisms. RECENT FINDINGS: There is good evidence for significant cognitive heterogeneity in schizophrenia. It is not yet clear, however, whether this heterogeneity is better accounted for by a general loss of function, varying in degree between different patients, or by impairment in specific cognitive abilities, for example working memory. Molecular genetic studies have provided evidence for associations of single nucleotide polymorphisms with both specific and general impairments, with some additional support for a working memory deficit from neuroimaging studies. SUMMARY: Larger, better controlled studies are needed before the genetic sources of cognitive heterogeneity in schizophrenia can be accurately characterized. This will be aided with the development and use of more specific neuropsychological tasks that can accurately discriminate between different cognitive domains. PMID- 17415082 TI - Magnetic resonance analysis of amygdalar volume in Alzheimer's disease. AB - PURPOSE OF REVIEW: With the rising prevalence of Alzheimer's disease, there is an increasing need for better comprehension of its pathophysiology. The purpose of this article is to review recent studies investigating the association between amygdalar volume and clinical symptoms in Alzheimer's disease. The first part describes the technique of MRI segmentation of amygdala. The advantages and risks of various segmentation techniques are noted. In the second part, the role of amygdalar volume in the assessment of clinical diagnosis is discussed. The third part encounters the relationship between the amygdalar atrophy and its neuropsychological correlates. RECENT FINDINGS: Numerous MRI studies showed the same degree of hippocampal and amygdalar volume loss. MRI volumetry of the amygdala may be relevant as a marker of dementia severity in Alzheimer's disease. Asymmetry in amygdalar atrophy is useful in separating Alzheimer's disease and frontotemporal lobar degeneration. There has been a lack of direct relationship between the atrophy of amygdala and neuropsychiatric symptoms in Alzheimer's disease. SUMMARY: Although time-consuming, the manual tracing represents the golden standard in MRI volumetry of amygdala. The pathogenesis of neuropsychiatric symptoms in Alzheimer's disease is complex and their manifestation is therefore not attributable to the amygdalar atrophy only. PMID- 17415083 TI - The neuropsychiatry of multiple sclerosis: a review of recent developments. AB - PURPOSE OF REVIEW: The aim of this review is to summarize the current literature on the neuropsychiatry of multiple sclerosis (MS). RECENT FINDINGS: Data from community samples have supported earlier findings from tertiary referral centres of high rates of depression in MS patients. Neuroimaging offers important clues as to the pathogenesis of depression, but psychosocial factors cannot be ignored and emerge as equally important predictors. Cognitive-behavioural therapy is an effective treatment, rivalling standard dosing of sertraline in patients with depression. An allied disorder--pseudobulbar affect--occurs in up to 10% of MS patients and responds well to a combination of dextromethorphan and quinidine. Cognitive dysfunction affects approximately 40% of MS patients. Markers of cerebral atrophy have emerged as more important correlates of impaired cognition than lesion volume. Moreover, functional MRI studies have demonstrated the brain's ability to compensate, in part, for damage. Should the disease burden be too severe, however, compensatory mechanisms fail and cognitive deficits increase accordingly. SUMMARY: Neuropsychiatric abnormalities are common in MS patients. No aspect of mentation is spared. Advances in neuroimaging are increasing our understanding of the pathogenesis of these disorders. Translating these findings into improved methods of treatment for patients presents researchers with pressing challenges. PMID- 17415084 TI - Bibliography. Current world literature. Addictive disorders. PMID- 17415085 TI - Cancer prevention: from public health interventions to individual tailoring. PMID- 17415086 TI - Cumulative cancer mortality risk and potential years of life lost to 64 years of age in Ireland, 1953-2002. AB - Premature cancer mortality trends were examined by reviewing cumulative mortality risk ('cumulative risk' hereafter) and potential years of life lost (PYLL) up to and including 64 years of age between 1953 and 2002 in Ireland. Trends were assessed quantitatively by Joinpoint analysis of both measures (with PYLL expressed as an age-standardized rate). The age of 64 years was used for these summary measures to reflect the focus of the Irish Government's cancer strategy on cancer in the under-65 population. Some differences emerged when ranking the significant types of cancer using cumulative risk and PYLL values. In general, however, the two methods generated similar overall trends, although PYLL rates tended to show steeper or longer-term declines, presumably reflecting the greater weight given to deaths at younger ages. Most cancers have, in recent years, shown a downward, or levelling-off of, trend for both sexes. The only exceptions were significant increases for oesophageal cancer in men (both measures), and prostate cancer (cumulative risk), cervical cancer (PYLL rate) and lymphoma in both sexes (cumulative risk). Rankings based on both cumulative risk and PYLL showed that male lung cancer is still the leading cause of premature death from cancer in Ireland, despite recent falls in mortality rates. Breast cancer has consistently been the leading cause of premature cancer death in women since the 1950s. Stomach cancer was once the second leading cause of premature cancer death in women, but since the 1960s it has been replaced by lung cancer. Ovarian cancer, having had a middle ranking for many years has, since the early 1990s, become the third leading cause of premature cancer death for women. PMID- 17415087 TI - Levels and correlates of alpha-fetoprotein in normal pregnancies among Caucasian and Chinese women. AB - Alpha-fetoprotein has been suggested to have anti breast cancer properties both in adult life and in utero. We studied correlates of maternal serum alpha fetoprotein levels in normal singleton pregnancies. This was a prospective study relying on women attending maternal units in major teaching hospitals in Boston, USA and Shanghai, China. Specifically, 212 Caucasian women in Boston, and 196 Asian women in Shanghai provided blood samples at the 16th and 27th gestational week. Maternal serum alpha-fetoprotein levels were measured and correlated with maternal and newborn parameters through multiple regression procedures, controlling for a set of potential confounders, including maternal levels of measured hormones. Alpha-fetoprotein was strongly inversely associated with maternal prepregnancy body mass index (-4.73 ng/l at the 27th week per 1 kg/m of body mass index, with 95% confidence intervals -7.09 to -2.36), whereas it was not related to parity, gender of offspring or birth weight. Duration of gestation was inversely associated with maternal alpha-fetoprotein levels, particularly among Caucasian women (-0.22 weeks per 60 ng/l of alpha-fetoprotein at the 27th week, with 95% confidence intervals -0.39 to -0.05). In normal pregnancies, maternal alpha-fetoprotein is inversely related to prepregnancy body mass index and appears to have a physiologic role on duration of gestation rather than on birth weight. PMID- 17415088 TI - Chemoprevention of lung cancers: lessons from CARET, the beta-carotene and retinol efficacy trial, and prospects for the future. AB - The objective of this paper was to review the strategies for lung cancer chemoprevention. A retrospective assessment of the major findings from the most informative lung cancer chemoprevention clinical trials [alpha-tocopherol (vitamin E), beta-carotene trial and beta-carotene and retinol efficacy trial] was employed. Both trials and many others showed no benefit from what was once the prime candidate for lung cancer chemoprevention, beta-carotene. Furthermore, both trials found that beta-carotene, alone or in combination with vitamin E or retinyl palmitate, increased the incidence of lung cancers and the total and cardiovascular mortality rates. In conclusion, design, conduct, documentation, relationships with participants, and preparedness for unexpected findings are all important for chemoprevention research. Trials are necessary to test inferences from observational epidemiology and animal models. Multiple classes of promising agents are available for evaluation and for eventual randomized trials. PMID- 17415089 TI - Host and environmental factors for gastric cancer in Babol, the Caspian Sea Coast, Iran. AB - To clarify host and environmental factors for gastric carcinogenesis, we obtained information about gastric cancer mortality in Babol, in the North of Iran, and recruited 130 participants aged 30-80 years from the general population of Babol in 2004. A urea breath test, assessment of IgG antibodies to Helicobacter pylori, a pepsinogen test, a marker of chronic atrophic gastritis, and determination of urinary excretions of sodium and potassium were performed. Diet and lifestyle information was also obtained using a questionnaire. The stomach cancer mortality rate for men in Babol (38.2/10(5)) was found to be somewhat lower than that for Japanese men (45.1/10(5)), while the mortality for women (26.9/10(5)) was higher than for Japanese women (20.9/10(5)). Positive rates for the urea breath test were 77.5 and 81.8% for Iranian men and women, respectively. Helicobacter pylori IgG antibodies were present in 68.7 and 73.7% of Iranian men and women, respectively, both values being marginally higher than for Japanese. We also found 51.0 and 52.8% to be positive for a pepsinogen test, significantly higher than the Japanese values. Urinary excretions of salt and potassium in this population appeared approximately the same as the consumption in Japanese. The elevated gastric cancer mortality in both men and women in Babol seems, by and large, to be related to higher H. pylori infection rates and prevalence of chronic atrophic gastritis. Certain factors, including H. pylori DNA diversity, host factors and their interactions, together with the level of medical practice, prevalence of and access to secondary prevention of stomach cancer, may also be associated with the relatively high mortality. PMID- 17415090 TI - Expression of CD147 as a significantly unfavorable prognostic factor in hepatocellular carcinoma. AB - The aim of this study is to investigate whether the expression of CD147 could be a prognostic factor for hepatocellular carcinoma. Tissue samples from 111 hepatocellular carcinoma patients were immunohistochemically stained with anti CD147, anti-matrix metalloproteinases-2 and anti-vascular endothelial growth factor antibodies. Tumor microvessel density was evaluated using CD34. The survival curves were estimated by Kaplan-Meier analysis and the prognostic significance of the marker was analyzed using the log-rank test. In addition, the identification of relevant prognostic factors was performed by multivariate Cox regression analysis. CD147 was mainly expressed in cancerous lesions and its expression was positively correlated with metalloproteinases-2 (P<0.0001), vascular endothelial growth factor (P<0.0001) and microvessel density CD34 (P<0.0001). Furthermore, CD147 was significantly associated with the presence of venous invasion (P=0.0013), tumor size (P<0.0001) and pTNM tumor stages (P=0.0001), as well as serum alpha-fetoprotein level (P<0.0001). Patients with positive expression of CD147 had poorer tumor recurrence-free survival than those with negative expression of CD147 (P<0.0001). Analyzed by a proportional hazard model, strong expression of CD147 had the highest risk ratio of recurrence among these markers (P<0.0001). The findings suggest that CD147 may be a significant independent predictor of poor survival in patients with hepatocellular carcinoma, and may be involved in tumor growth, invasion and angiogenesis in hepatocellular carcinoma. PMID- 17415091 TI - Translational approaches for the prevention of estrogen receptor-negative breast cancer. AB - Breast cancer prevention has focused heavily on endocrine interventions using selective estrogen receptor modulators and aromatase inhibitors. Tamoxifen, the stereotypical selective estrogen receptor modulator, significantly reduces the breast cancer incidence in high-risk women. Selective estrogen receptor modulators and aromatase inhibitors, however, only prevent the development of estrogen receptor-positive breast cancer and have no effect in reducing the risk of estrogen receptor-negative breast cancer, which has poor prognosis. Thus, preventive therapies for estrogen receptor-negative breast cancer are clearly needed. Recently, a number of novel chemopreventive agents targeting nonendocrine pathways have been developed and shown to prevent estrogen receptor-negative mammary tumorigenesis in animal models. These agents include rexinoids, selective cyclooxygenase-2 inhibitors, tyrosine kinase inhibitors, and others. In this review, we discuss the effects of selective estrogen receptor modulators and aromatase inhibitors, as well as novel agents targeting nonendocrine pathways. We also discuss the promise of combining these agents for the effective prevention of all forms of breast cancer. PMID- 17415092 TI - Evaluation of mathematical models for breast cancer risk assessment in routine clinical use. AB - Chemoprevention, prophylactic surgery, and intensified screening can be offered to patients with an increased lifetime risk, p(life), for breast cancer. Estimation of p(life) includes BRCA analysis and risk estimation based on individual risk factors and family history. MENDEL and BRCAPRO are models that estimate the probability of BRCA1/2-mutations, p(mut), and p(life). In this study, the models are compared with Ford and Claus penetrance/frequency functions. The results were compared with the Tyrer-Cuzick model. Genetic analysis of 111 breast cancer-affected patients from 103 kindreds with a family history of breast and/or ovarian cancer (German Consortium for Hereditary Breast and Ovarian Cancer) was carried out by sequencing BRCA1 and BRCA2. p(life) and p(mut) were calculated with MENDEL, BRCAPRO(Claus), BRCAPRO(Ford), as well as the Tyrer-Cuzick model. The accuracy of p(mut) was analyzed by receiver operating characteristics, and p(life) of each model was compared. The strongest correlation of p(life) was shown by BRCAPRO(Ford)/MENDEL, at r=0.69; no correlation was shown by BRCAPRO(Claus)/MENDEL, at r=0.018. The Tyrer-Cuzick model had the strongest correlations with MENDEL and BRCAPRO(Ford). For MENDEL and BRCAPRO, low correlation or p(mut)-prediction was improved by excluding kindreds with ovarian cancer. p(mut) showed the best accuracy for BRCAPRO(Ford) and MENDEL. BRCAPRO and MENDEL are useful tools for calculating p(mut). They can provide support in decision-making for or against genetic analysis. Estimations of p(life) and p(mut) based on a mathematical model should use algorithms and penetrance/frequency data appropriate to the population counseled. Reproductive/hormonal data, should be incorporated as Tyrer-Cuzick does. PMID- 17415093 TI - Evaluation of effectiveness of quality-assured mammography screening in Germany: sample size considerations and design options. AB - In cancer screening, it is considered mandatory not only to prove the efficacy of the screening test, but also, as a permanent commitment, to demonstrate and quantify the effectiveness of service screening in terms of mortality reduction when the test becomes part of a screening programme. In Germany, a nationwide organized and quality-assured mammography screening programme among 50-69-year old women, a target population of about 10 million women, is currently implemented. The envisaged short implementation period of about 2 years raises the issue of how to estimate the effectiveness of the programme in terms of long term mortality reduction. On the basis of sample size calculations, a strategy for evaluation of mortality reduction is outlined. Our approach compares breast cancer mortality in different geographical areas or temporally within geographical areas. The latter design appears useful also for established programmes to examine site-specific mortality reduction by screening in late starting areas, which are frequently used as control areas of a geographical comparison, but rarely a subject of evaluation on their own. Emphasis has to be put on the collection of the required data from the very beginning of the programme. For this, performance of cancer registration and proper linkage with the screening programme must be enhanced. In the trade-off between data protection and public interest in quality-assurance of medical care, emphasis should be put on assurance of high-level medical care whose relevance may be counted in terms of the numbers of deaths prevented. It is from this the individual truly benefits. PMID- 17415094 TI - Hormonal interventions to prevent hormonal cancers: breast and prostate cancers. AB - In 1998, the concept of breast cancer prevention became a reality with the approval of tamoxifen to reduce the risk of developing breast cancer in women at increased risk for the disease. This approval was based on decades of research on selective estrogen receptor modulators providing an understanding of the role of the estrogen receptor in breast cell growth, and an appreciation of the carcinogenic process. Although results from the Breast Cancer Prevention Trial demonstrated a 49% reduction in breast cancer in women at increased risk, there were associated toxicities related to the estrogenic effects of tamoxifen; that is, deep vein thrombosis, pulmonary embolism, and endometrial cancer. In an effort to improve its benefit-risk profile, tamoxifen is now being compared with raloxifene, a selective estrogen receptor modulator approved for the treatment and prevention of osteoporosis. This equivalency prevention Study of Tamoxifen and Raloxifene completed accrual of 19 747 high-risk postmenopausal women in November 2004. Meanwhile, another class of estrogen-directed drugs, the aromatase inhibitors, have shown efficacy in breast cancer adjuvant trials, spawning a number of prevention trials that have recently been initiated. As with breast cancer the hormonal contribution to prostate carcinogenesis was the basis for the Prostate Cancer Prevention Trial which showed that finasteride, an androgen antagonist, reduces the incidence of prostate cancer compared to placebo. PMID- 17415095 TI - Use of electric blankets and association with prevalence of endometrial cancer. AB - The objective of this study was to assess the relationship between electric blanket use and prevalence of endometrial cancer for women. Information relating to women enrolled in the Women's Health Initiative Observational Data Set (n=93 676) used to test the relationship factors associated with endometrial cancer included older age at screening, younger age at last menstrual period, region of domicile (highest prevalence in the South), less than a high school education, lower income, body mass index >25 kg/m, low parity, unopposed use of estrogen, never use of estrogen plus progesterone, past alcohol use, higher percentage of daily calories from fat and any electric blanket use. Following a univariate identification of factors significantly related to endometrial cancer, stepwise logistic regression analysis was performed for those factors with P values of less than 0.001 in the univariate analysis. Using electric blankets was associated with a 15% higher prevalence of endometrial cancer than never having used electric blankets (odds ratio=1.15, 95% confidence interval: 1.03-1.27). After controlling for variables significantly associated with endometrial cancer, use of electric blankets for 20 years or more was associated with 36% higher prevalence of endometrial cancer (odds ratio=1.36, 95% confidence interval: 1.16 1.59). Although we were unable to determine the duration of electric blanket use before diagnosis of endometrial cancer, we found that women using electric blankets for 20 years or more had a significantly higher prevalence. PMID- 17415096 TI - Assessment of risk factors for oral squamous cell carcinoma in Chidambaram, Southern India: a case-control study. AB - Oral squamous cell carcinoma, the fifth most common cancer worldwide, is a major cause of morbidity and mortality in India. The effect of lifestyle factors, including tobacco chewing, smoking and alcohol drinking, diet and dental care, on the risk of oral cancer was investigated in a case-control study conducted in Rajah Muthiah Dental College and Hospital, Annamalainagar, Annamalai University, Chidambaram, Tamil Nadu, India during the period 1991-2003. The study included 388 oral squamous cell carcinoma cases and an equal number (388) of age and sex matched controls. All participants were interviewed using a structured questionnaire that contained data on demographic factors, family history of cancer, tobacco habits, use of alcohol, frequency, duration, cessation of these habits, dietary practices and oral hygiene. The data were analysed using multiple logistic regression model. Among people with chewing habits, those who chewed betel quid with tobacco [odds ratio (OR) 3.19, 95% confidence interval (CI): 0.48 2.13] and tobacco alone (OR 2.89) showed a greater risk than controls. Bidi smoking (OR 4.63) and alcohol drinking (OR 1.65) emerged as significant risk factors for oral cancer. These three habits showed increasing risk with increasing frequency and increase in duration of habits. Addition of alcohol to other habits also enhanced the risk for oral cancer. The combination of chewing and smoking together with alcohol drinking showed very high relative risk (OR 11.34). A positive association was observed between non-vegetarian diet, poor oral hygiene and poor dentition with the risk of oral squamous cell carcinoma. The fact that these risk factors are modifiable emphasizes the need for increasing awareness among the general public and policy makers as a first step in the prevention and control of oral squamous cell carcinoma. PMID- 17415097 TI - Access point analysis in smoking and nonsmoking adolescents: findings from the European Smoking Prevention Framework Approach study. AB - This study analyzed possibilities to access European adolescents for tobacco control activities in out-of-school settings as part of comprehensive tobacco control programs. Data on leisure time behaviors of secondary school students were gathered during three waves from six European Union countries as part of the European Smoking Prevention Framework Approach study. The results revealed that streets, shops, sport clubs, swimming pools and snack bars were the most popular leisure time settings for adolescents across the six countries. In addition, leisure time activities were similar across the six countries. European-wide prevention programmes can reach adolescents in similar out-of-school settings. Furthermore, smokers and nonsmokers differed in the places that they visited. Whereas smokers preferred social places such as bars, house/rave parties, discos, snack bars, streets and shops, nonsmokers preferred sport clubs and libraries. Smokers tended to visit more unsupervised and less structured places where smoking tends to be permitted, whereas nonsmokers tended to visit places with more structured activities and places under adult supervision. Over time, both smoking and nonsmoking adolescents tended to spend more of their leisure time in settings with low adult supervision. As structured activities under the supervision of adults can have a protective effect against the involvement in risky behaviours, participation in these types of activities must be promoted. Additional strategies for settings with less supervision need to be developed as they attract more smoking adolescents and older nonsmoking adolescents. PMID- 17415098 TI - Passive cancer detection and medical help seeking for cancer symptoms: (in)adequate behavior and psychosocial determinants. AB - The present study explored the performance and psychosocial determinants of passive cancer detection behavior, that is, attentiveness to cancer symptoms, and medical help seeking, and investigated potential dependency between these two behaviors. A detailed telephone survey was conducted among 459 respondents, aged 55 years or older. The survey assessed passive detection behavior and appropriately timed medical help seeking, regarding 14 cancer symptoms. Knowledge of cancer symptoms and various other psychosocial determinants were also measured. Knowledge of cancer symptoms and adequate passive detection behavior was low to moderate. Timely medical help-seeking behavior was low to moderate for urgent symptoms but relatively high for prolonged symptoms. Overall, women had higher knowledge levels, paid more attention to cancer symptoms, and performed more timely help seeking than men. Passive detection behavior was positively associated with premotivational awareness factors (knowledge and awareness), female gender, and perceived advantages. Timely medical help seeking was positively related to cognitive motivational factors (perceived advantages, self efficacy expectations, and intention) and negatively related to educational level and perceived susceptibility to cancer. Furthermore, a strong positive association was found between the performance of passive detection behavior and timely medical help seeking. The suboptimal levels of knowledge and performance of early cancer detection behaviors found in this study emphasize a need for educational efforts in the area of early cancer detection. The effectiveness of these efforts may benefit from considering the distinct sets of determinants of passive detection behavior and medical help seeking. PMID- 17415099 TI - Transforming the care delivery model in preparation for the clinical nurse leader. PMID- 17415100 TI - Shortage strategies: retaining the experienced nurse. PMID- 17415101 TI - Hospital billing and reimbursement: charging for inpatient nursing care. PMID- 17415102 TI - Competencies and tips for effective leadership: from novice to expert. PMID- 17415103 TI - Recruit, retain, and recognize with a professional nurse ambassador program. PMID- 17415104 TI - Shared governance: making the transition in practice and perception. AB - The shared governance literature contains numerous examples of how to design and implement nursing shared governance models. However, there is a major gap between design/implementation and a change in culture. A change in nursing culture will support viability of this governance model. The authors detail the steps taken by a shared governance transition team to help a large nursing organization make changes in governance process and perception as well as to incorporate a maintenance plan. PMID- 17415105 TI - The role of information technology in healthcare communications, efficiency, and patient safety: application and results. AB - The authors document the development and deployment of technology that serves to enhance healthcare communications, efficiency, and patient safety at an 881-bed, level 2 trauma center. This technology includes a patient-centric call process using real-time, rules-based task assignments and tracking software that guides personalized and timely response to the individual patient' needs and allows the mining of data for outcome analysis. PMID- 17415106 TI - Failure to rescue: a literature review. AB - Rapid response teams have been advocated as an intervention to reduce failure to rescue events. Such teams can improve nurse autonomy and control to rescue patients deteriorating in a medical surgical setting. The purpose of this review is to enhance nurse executives' understanding of failure to rescue as a nurse sensitive outcome, tested interventions, and implications for future research. The emergence of failure to rescue as an outcome measure will be initially discussed. Research regarding the relationship between failure to rescue and registered nurse staffing as well as research examining the potential to reduce failure-to-rescue events will be explored. PMID- 17415107 TI - Nursing support, workload, and intent to stay in Magnet, Magnet-aspiring, and non Magnet hospitals. AB - This study examined the differences between nurses' (N = 3,337) scores on organizational support, workload, satisfaction, and intent to stay between Magnet, Magnet-aspiring, and non-Magnet hospitals. The study was conducted using the Individual Workload Perception Scale, a valid and reliable tool with 32 Likert scale items, with nurses from 11 states, 15 institutions, and 292 diverse units. Results indicate that nurses at Magnet hospitals had significantly better scores on all subscales. Furthermore, nurses from Magnet-aspiring hospitals had better scores than did nurses from non-Magnet facilities. Conclusions of the study indicate that the Magnet program is meeting its intended goal: to provide a professional practice environment for staff nurses. Nurse executives may consider using the Individual Workload Perception Scale as a way to assess their organization's culture as it relates to professional practice of the registered nurse. PMID- 17415108 TI - The use of p16 as a marker for cervical cancer precursor lesions. PMID- 17415109 TI - Diagnostic loop electrosurgical excisional procedure for discrepancy: do preoperative factors predict presence of significant cervical intraepithelial neoplasia? AB - OBJECTIVE: Although pathological discrepancy between Pap smear and biopsy is an accepted indication to perform a diagnostic loop electrosurgical excision procedure (LEEP), this procedure is not without complications. Our objective was to determine the incidence of cervical intraepithelial neoplasia (CIN) 2,3 and patient factors that increase the likelihood of detecting CIN 2,3. MATERIALS AND METHODS: We performed a retrospective chart review of patients who underwent a diagnostic LEEP for pathological discrepancy at a university-based colposcopy clinic. Pathological discrepancy is defined as a high-grade Pap smear with a colposcopically directed biopsy of CIN 1 or less. Demographic, cytological, and histological information were collected using a computerized database. The patients were divided into 2 groups (CIN 2,3 and CIN 1 or less) based on the pathology from the LEEP specimen. Patient factors were compared with final pathological results using chi(2) test, Student t test, Wilcoxon rank sum test, and multivariate analysis as indicated. RESULTS: A total of 102 patients were identified. Seven patients had normal specimens, 3 had HPV changes, 25 had CIN 1, 29 had CIN 2, and 38 had CIN 3. Thirty-five patients (34%) had CIN 1 or less, whereas 67 patients (66%) had CIN 2,3. The 2 groups were comparable in terms of age (30.4 vs 28.1 years), parity (2.2 vs 1.9), and age of coitarche (16.3 vs 16.4 years). No statistical difference existed between the groups regarding race, smoking status, Pap smear, history of previous cytological abnormality, contraception method, number of previous sexual partners, and HIV status. The majority of patients (75%) had not undergone previous treatment of CIN. The CIN 2,3 group were more likely than the CIN 1 or less group to have had previous treatment or biopsy for CIN (66% vs 34%; p = .004). Univariate (p = .004) and multivariate (p < .001) analysis demonstrated previous treatment of CIN as the only significant factor predicting CIN 2,3. CONCLUSION: Two thirds of women undergoing a LEEP for pathological discrepancy between Pap smear and cervical biopsy will have CIN 2,3. Women that have had previous treatment of CIN are more likely to have CIN 2,3 detected on their LEEP specimen. PMID- 17415110 TI - Comparison of human papilloma virus testing and spectroscopy combined with cervical cytology for the detection of high-grade cervical neoplasia. AB - OBJECTIVE: This study compared the performance of cervical cytology plus human papilloma virus testing (Pap + HPV) or cervical spectroscopy (Pap + CS) for identifying high-grade cervical neoplasia in a high-risk population of women referred for colposcopy. MATERIALS AND METHODS: Each of 113 subjects underwent spectroscopy, thin-layer cytology, HPV testing, colposcopy, biopsy when indicated, and/or endocervical curettage. Evaluable data for analysis were collected for 102 of the subjects. Sensitivity and specificity were calculated for both strategies. RESULTS: Pap + HPV and Pap + CS achieved equivalent sensitivities (95%) for high-grade lesions, with both detecting 17 of 18 histology confirmed cervical intraepithelial neoplasia (CIN) 2+ lesions. Pap + HPV had a specificity of only 27.4% compared with 65.5% for Pap + CS (p < .0001). CONCLUSIONS: Spectroscopic interrogation of the cervix is equally sensitive and 2 fold more specific than HPV testing when combined with cervical cytology for identifying high-grade cervical neoplasia. PMID- 17415111 TI - Cyclooxygenase 2 expression in cervical intraepithelial neoplasia and vulvar cancer. AB - OBJECTIVE: The purpose of this study is to describe the expression of cyclooxygenase (COX) 2 cervical intraepithelial neoplasia (CIN) and vulvar cancer specimens. MATERIALS AND METHODS: Archived tissues from 21 cases including 6 cases negative for CIN (no CIN), 71 low-grade (CIN 1) diseases, 8 high-grade (CIN 2, 3) diseases, and 14 vulvar cancer cases were examined. Immunohistochemistry was evaluated in COX-2 expression in tissue using an isoform-specific COX-2 polyclonal antibody. Specimens were assigned an immunohistochemical score for intensity of staining and the percent of cells stained. The slides were scored by 2 independent pathologists and compared across histological categories. RESULTS: A greater proportion of cells were stained in specimens with high-grade CIN (p = 0.01). Staining intensity was not statistically different among the 3 groups. Higher scores were found for vulvar cancer as compared with normal vulva (p = 0.01). CONCLUSIONS: The increase in COX-2 in cervical cancer precursors may provide a potential target for prevention studies. PMID- 17415112 TI - Is atypical squamous cells that cannot exclude high-grade squamous intraepithelial lesion clinically significant? AB - OBJECTIVE: To determine the cumulative risk of cervical intraepithelial neoplasia (CIN) 2 or 3 in patients with atypical squamous cells, cannot exclude HSIL (ASC H). METHODS: A retrospective analysis was performed to identify patients referred to the dysplasia clinic with ASC-H. Initial evaluation included colposcopy, endocervical curettage, and an ectocervical biopsy, when indicated, in all the patients. A follow-up evaluation was performed at 6 and 12 months. Cumulative histological diagnosis of CIN 2 or 3 at 12 months served as the clinical end point. RESULTS: Two hundred twenty-nine patients with ASC-H and with a mean age of 32.8 years were evaluated. At the time of initial colposcopy, only 10.0% (23/229; 95% CI = 6.5%-15%) of the patients had histological evidence of CIN 2 or 3. The cumulative risk of CIN 2 or 3 was 12.2% (95% CI = 8%-17%). CONCLUSIONS: Evaluation of patients with ASC-H with colposcopy does lead to the detection of CIN 2 or 3 but perhaps at a rate lower than previously reported. PMID- 17415113 TI - Outcomes after treatment of cervical intraepithelial neoplasia among women with HIV. AB - OBJECTIVE: To describe outcomes after treatment of cervical intraepithelial neoplasia (CIN) in women with HIV. MATERIALS AND METHODS: Women in two prospective cohort studies, the Women's Interagency HIV Study (WIHS) and the HIV Epidemiology Research Study (HERS), were followed every 6 months after treatment of CIN using human papillomavirus (HPV) testing and cytology with colposcopy as indicated. Identification of CIN or a squamous intraepithelial lesion (SIL) within 6 months was defined as treatment failure and later disease as recurrence. RESULTS: Follow-up was available for 170 HIV-seropositive and 15 HIV-seronegative women. Treatment failed in 84 (45%) women (79 HIV seropositive and 5 HIV seronegative). Failure was more likely in women with lower CD4 counts (CD4 < 200 cells/microL: odds ratio [OR] = 2.96; 95% CI = 1.4-6.2) and detectable HPV DNA (OR 8.20; 95% CI = 1.8-37.4; p = .01). After successful treatment, recurrence free probabilities at 1,2, 3, and 5 years were .79, .64, .49, and .34, respectively. HIV-seronegative women were less likely to recur than HIV seropositive women (p = .03). In multivariable analysis of HIV-positive women, recurrence was more likely among women treated for CIN 2,3 (hazard ratio [HR] = 2.4; 95% CI = 1.4-4.8), those with CD4 count of less than 200 cells/microL (HR = 2.9; 95% CI = 1.3-6.5) and those with HPV after treatment (HR 2.9; 95% CI = 1.4 6.1); oncogenic HPV was more strongly associated with recurrence than nononcogenic HPV (p(trend) = .009). Most failures and recurrences were low grade, but one adenocarcinoma was diagnosed 4.2 years after therapy for CIN 1. CONCLUSION: Treatment failure and recurrence are common in women with HIV but are usually low grade. PMID- 17415114 TI - Immunohistochemical expression of p16(INK4A) in normal uterine cervix, nonneoplastic epithelial lesions, and low-grade squamous intraepithelial lesions. AB - OBJECTIVES: In this study, the authors analyzed the immunoexpression of p16 in high-risk human papillomavirus DNA-negative normal and nonneoplastic cervical epithelia, in low-grade cervical intraepithelial neoplasia (CIN), high-grade CIN, and squamous cell carcinoma. MATERIALS AND METHODS: A retrospective study, in which 58 normal cervical hysterectomy samples, 56 nonneoplastic cervical biopsies, 88 CIN 1, 33 CIN 2, 32 CIN 3, and 47 invasive squamous cell carcinoma biopsies, were evaluated for p16 immunoexpression. Human papillomavirus tests were also performed. RESULTS: p16 immunohistochemistry seems to reveal possible different biological subgroups of lesions among morphologically similar mildly dysplastic cervical epithelia. CONCLUSION: Distribution patterns of p16 protein might be useful to predict different outcomes in CIN 1. PMID- 17415115 TI - Primary spindle cell sarcoma of the vagina treated with neoadjuvant radiation and pelvic exenteration. AB - OBJECTIVE: Malignant neoplasms of the vagina are rare gynecologic tumors. Primary vaginal sarcomas are even more unusual lesions, representing fewer than 2% of malignant vaginal lesions. CASE: We present a case of a primary vaginal spindle cell sarcoma, treated with neoadjuvant radiation followed by total pelvic exenteration. The patient remains without evidence of disease 2 years after surgery. CONCLUSIONS: The mainstay of treatment of vaginal sarcomas is surgical. Neoadjuvant radiation treatment may decrease surgical morbidity and lead to long term cure. PMID- 17415116 TI - Auditing the use of colposcopy versus general gynecology clinics to investigate women with postcoital or intermenstrual bleeding: a case for a new outpatient service. AB - OBJECTIVE: The aim of this audit was to determine whether initial referral to a general gynecology clinic (GOPD) or a colposcopy clinic is the most efficient means of managing women with postcoital (PCB) or intermenstrual (IMB) bleeding. MATERIALS AND METHODS: A prospective audit of women with PCB or IMB was conducted. Sixty referrals from primary care were note-logged and alternatively allocated to either clinic. Data were collected by case note review. RESULTS: Sixty women were referred for abnormal bleeding, 33 (55%) of which were seen at the GOPD and 27 (45%) at the colposcopy clinic. Investigations included Chlamydia testing, which was deficient in both settings, whereas ultrasound scans and hysteroscopies were performed in 9% of patients attending the GOPD compared with none in the colposcopy clinic. The most common finding at the GOPD was cervical ectopy in 14 (42%) patients. No abnormality was found in 9 (28%) patients. At colposcopy, 14 (52%) had no abnormality detected, whereas 8 (30%) had cervical ectopy and 3 (11%) had cervical intraepithelial neoplasia. There were no cases of malignancy found in this study. CONCLUSION: Most patients with PCB and IMB will not have a serious abnormality. Our study proposes that a new PCB/IMB outpatient service can improve the efficiency of treatment of these women by providing appropriate guidelines to standardize their care. PMID- 17415117 TI - Giant cell fibroblastoma of the vulva at the site of a previous fibroepithelial stromal polyp: a case report. AB - BACKGROUND: Giant cell fibroblastoma (GCF) is an unusual soft tissue tumor, occurring predominantly in infants and children, and rarely in adults. Giant cell fibroblastoma develops de novo in the dermis or subcutis with a predilection for the extremities, the abdominal and chest walls, umbilical and inguinal regions. CASE: A GCF arose at the same site (labium majus of vulva) as a previous cellular fibroepithelial stromal polyp in a 28-year-old woman. CONCLUSION: We report a case of GCF of the vulva, an unreported site. PMID- 17415118 TI - Surgical resection of vulvar metastases of endometrial cancer: a presentation of two cases. AB - OBJECTIVE: Endometrial cancer generally carries a good prognosis. However, 10% to 15% of patients will manifest recurrent disease. One half of these recurrences are confined to the vagina. Whereas pelvic recurrence is most common in patients who do not receive postoperative adjuvant radiation therapy, distant metastases predominate among patients who received postoperative radiation therapy. Surgical resection of disease may be possible, therapeutic and even curative, in select patients with isolated cancer recurrence. CASE 1: A 63-year-old patient presented 7 years after treatment of endometrial cancer with a vulvar lesion and groin mass. The lesions were successfully resected and confirmed to be recurrent endometrial cancer. Adjuvant radiation and chemotherapy were prescribed leading to a complete clinical response. This patient survived without evidence of disease for 1 year. However, she eventually died 8 months later because of a disease recurrence. CASE 2: An 83-year-old patient with a history of a hysterectomy for endometrial cancer and radiation therapy for a vaginal vault recurrence presented with an exophytic labial mass. After radical wide excision of her vulvar mass and bilateral groin dissection, final pathology revealed that the mass was consistent with recurrent endometrial cancer. This patient remains without evidence of disease 18 months after treatment of disease recurrence. CONCLUSIONS: Uncommon sites of recurrence of endometrial cancer may include the vulva. These rare metastases may be amenable to surgical resection with adjuvant therapy as indicated. PMID- 17415119 TI - Clinical question: ask the expert. PMID- 17415120 TI - Home study course: spring 2007. AB - OBJECTIVE: The Home Study Course is intended for the practicing colposcopist or practitioner who is seeking to develop or enhance his/her colposcopic skills. The goal of the course is to present colposcopic cases that are unusual or instructive in terms of appearance, presentation, or management, or that demonstrate new and important knowledge in the area of colposcopy or pathology. Participants may benefit from reading and studying the material or from testing their knowledge by answering the questions. ACCME ACCREDITATION: The American Society for Colposcopy and Cervical Pathology (ASCCP) is accredited by the Accreditation Council for Continuing Medical Education (ACCME) to provide continuing medical education for physicians. The ASCCP designates this education activity for a maximum of 1 AMA PRA Category I Credit trade mark. Physicians should only claim credit commensurate with the extent of their participation in the activity. The ASCCP also designates their educational activity for 1 Category 1 credit hour of the ASCCP's Program for Continuing Professional Development. Credit is available for those who choose to apply. The Home Study Course is planned and produced in accordance with the ACCME's Essential Areas and Elements. DISCLOSURE: The clinical history and images in the Home Study Course may represent an actual case, but not always. To improve educational quality, some gross, cytological, or histological images may come from photographic libraries. Good teaching cases are often difficult to obtain, and we encourage our readers to submit cases with high-quality images to the Home Study Course editor or executive editor to consider for publication. Lastly, faculty must disclose any significant financial interest or relationship with proprietary entities that may have a direct relationship to the subject matter. For this course, the authors had the following relationships to report: Kevin J. Mitchell, MD: Digene Corporation: Speaker: Honorarium; Stockholder Cytyc Corporation: Stockholder 3M Corporation: Phase 2 Drug Testing: Research funding. Lisa Flowers, MD: No such relationship to report ACKNOWLEDGMENT: Cytology and histology courtesy of Dennis O'Connor, MD. PMID- 17415123 TI - Practice patterns, safety, and rationale for tracheostomy tube changes: a survey of otolaryngology training programs. AB - INTRODUCTION: Tracheotomy for long-term ventilation is a common surgical procedure in the hospital setting. Although the postoperative care is often perceived as routine, complications associated with tracheostomy changes may result in loss of airway and death. In addition, the practice patterns, rationale, and complications related to tube changes have been poorly described. STUDY DESIGN AND METHODS: A survey of chief residents in accredited otolaryngology training programs was performed to determine the management strategies, rationale, and complications associated with postoperative tracheostomy tube changes. RESULTS: The first tube change was performed after a mean of 5.3 (range, 3-7) days after the procedure, most frequently by junior residents. The first change was performed in a variety of locations including the intensive care unit (88%), step down unit (80%), and regular floor (78%). Twenty five percent performed these changes at night or on weekends. The most frequently reported rationale for performing routine tracheotomy changes was examination of the stoma for maturity (46%), prevention of stomal infection (46%), and confirmation of stability for transport to a less monitored setting (41%). Twenty five (42%) respondents reported awareness of a loss of airway, and nine (15%) respondents reported awareness of a death as a result of the first tube change at their institution during their residency. A statistically significant higher incidence of airway loss was reported by respondents who reported performing the first tube change on the floor (96.1% vs. 63.6%). CONCLUSION: There is significant variability in the approach to postoperative tracheostomy tube management. The occurrence of major complications including deaths from routine tube changes requires an examination of the rationale and safety of this practice. PMID- 17415124 TI - Flexible laryngoscopy-guided pharyngeal pH monitoring in infants. AB - OBJECTIVE: To evaluate flexible laryngoscopy-guided pharyngeal pH probe monitoring as an alternative accurate and practical pharyngeal pH probe placement technique to eliminate the need for radiographs and esophageal manometry for pharyngeal pH monitoring in infants with extraesophageal reflux disease (EERD). MATERIALS AND METHODS: Infants with suspected laryngopharyngeal acid reflux (LPR) who underwent pharyngoesophageal pH monitoring were included. Data analysis included description of the flexible laryngoscopy-guided pharyngoesophageal pH monitoring technique and pharyngoesophageal pH monitoring findings, including the number of acid reflux episodes and percent acid exposure time. RESULTS: Six infants (3 boys, 3 girls, age range, 2 wk-7.5 mo) with suspected LPR underwent pharyngoesophageal pH monitoring. Flexible laryngoscopy was used, under direct vision, to guide pH probe placement in the laryngopharyngeal region. The esophageal pH probe was located 5 cm distal to the pharyngeal pH probe. All infants tolerated the procedure. Of the six infants, four had pharyngeal acid reflux, and six had esophageal acid reflux. The number of acid reflux episodes ranged from 4 to 81 in the pharynx and from 5 to 173 in the esophagus. The percentage of acid exposure time was between 0% and 1.2% in the pharynx and between 0.1% and 1.5% in the esophagus. CONCLUSION: With the aid of flexible laryngoscopy, a pH probe can be placed in the laryngopharyngeal region in infants undergoing pharyngeal pH monitoring. Findings documented that not all esophageal acid reflux reach the pharynx. Flexible laryngoscopy-guided pharyngeal pH probe placement can be used to detect LPR in infants with EERD. PMID- 17415125 TI - Quantifying the physiology of laryngotracheal stenosis: changes in pulmonary dynamics in response to graded extrathoracic resistive loading. AB - OBJECTIVES/HYPOTHESIS: A model of airway obstruction was developed to study the impact of changes in airway resistance on ventilatory mechanics. This was used to derive quantitative indices of airway obstruction to aid in the objective diagnosis and physiological monitoring of adult patients with laryngotracheal stenosis (LTS). METHODS: Six airway resistors, the characteristics of which mirrored the selective impediment to inspiratory airflow that occurs in patients with LTS, were created and calibrated. Maximum-effort flow-volume loops were obtained from 15 volunteers with resistors placed in series with the spirometer. Diagnostic and monitoring performances of various flow-volume indices were assessed with receiver-operating characteristics (ROC) and analysis of variance (ANOVA), respectively. Promising indices were further evaluated in patients with LTS. RESULTS: Experimentally, the ratio of expiratory and inspiratory flows at midvital capacity (MEF50/MIF50) and peak expiratory to inspiratory flow ratio (PEF/PIF) had diagnostic sensitivities of 87% and 89%, respectively. The best index, both experimentally and clinically, was the ratio of area under the expiratory and inspiratory curves (ratio of integrals), with experimental and clinical sensitivities of 97% and 100%, respectively. A clinical specificity of 95%, and area under the curve of 0.965 were achieved for this index. Both PEF/PIF and the ratio of integrals could identify step changes in airway resistance greater than 10 cmH2O . sec . L (P < .05; ANOVA). CONCLUSIONS: Flow-volume testing is simple and noninvasive and can be used to quantify the diagnosis and physiological monitoring of patients with LTS. The ratio of areas under the expiratory and inspiratory curves appears to be the optimal index for this purpose. PMID- 17415126 TI - Extraorbital skull base idiopathic pseudotumor. AB - OBJECTIVES/HYPOTHESIS: The term idiopathic pseudotumor (IP) refers to a nonspecific, nonneoplastic inflammatory process without identifiable local or systemic causes, which is one of the most common causes of intraorbital space occupying lesions. Occasionally, orbital pseudotumors may extend to other areas of the skull base. Rarely, pseudotumors may present as a skull base mass with no involvement of the orbit. The ophthalmology literature has detailed reviews of IP as an intraorbital space-occupying lesion, but lesions involving only the skull base and sparing the orbit are rare in the literature. We present a review of our experience with six patients with extraorbital skull base pseudotumor and a review of the relevant literature. METHOD: Retrospective case study. RESULTS: We treated six patients with extraorbital skull base pseudotumor between 1996 and 2004. Four patients had lesions in the pterygopalatine and/or infratemporal fossae, and two patients had lesions in the superior aspect of the parapharyngeal space. Five patients had excellent initial response to steroids. Two of the pseudotumors recurred. One patient partially responded to steroids and is currently stable with limited disease. Our literature review demonstrated 16 previously published reports of IP of the skull base sparing the orbit. CONCLUSIONS: A well-documented intraorbital lesion, inflammatory pseudotumor may present outside the orbit. Treatment of IP of the skull base is controversial and may involve corticosteroids or surgical resection, or both. Other chemotherapeutic agents and radiotherapy may be considered in steroid-resistant patients. Pathological subtype, ease and safety of resection, safety of high-dose corticosteroid use, the surgeon's comfort, and the patient's preference must be included in the decision-making process for treatment. PMID- 17415127 TI - Concentration and distribution of hyaluronic acid in human vocal folds. AB - OBJECTIVE/HYPOTHESIS: To evaluate the concentration and distribution of hyaluronic acid (HA) in human adult vocal folds. STUDY DESIGN: Experimental study. METHODS: Histologic samples of the anterior, middle, and posterior regions of the vocal folds were studied to determine the tissue distribution of HA. The HA was traced with hyaluronan-binding proteins (HAPB) from bovine cartilage. Chemical bonds were detected through the avidin-biotin immunoperoxidase method. A control with the specific Streptomyces hyalurolyticus hyaluronidase was conducted to avoid false-positive results. The concentration levels of HA were determined through a fluorescence-based assay method with the same HAPB and europium-labeled streptavidin. RESULTS: Women presented higher HA concentrations than men in every studied region of the vocal folds and overall HA concentration levels twice as higher. In regard to the distribution of HA, the macula flavae and the vocal fold intermediate and deep layers presented marked staining, with moderate staining around the vessels, on the gland capsules, and the connective tissue surrounding the vocal fold muscular fibers. CONCLUSIONS: HA is diffusely distributed along the vocal folds; the profuse amounts of HA observed in the female vocal folds may be related to impact absorption, acting as a protective factor against the characteristic high vibration frequency. PMID- 17415128 TI - A complex muscle fiber network in the cricothyroid muscle: a scanning electron microscopic study. AB - OBJECTIVES: To examine the three-dimensional ultrastructure of cricothyroid (CT) muscle fibers to elucidate their morphologic characteristics with regard to the specific functions of the muscle. STUDY DESIGN: An anatomic animal study. METHODS: The CT muscles of five adult rats were processed using the HCl hydrolysis method to remove the peri- and intramuscular connective tissue components. The fine muscle fiber arrangements were observed by scanning electron microscopy. RESULTS: Complex muscle fiber interconnections (myomyous junctions) were identified in the muscles. Myomyous junctions were characterized by the connection of many villous processes that were 1 to 2 mum in diameter and 2 to 4 mum in length with a serrated appearance at the ends of a lateral branch or a bifurcating trunk of the muscle fibers. CONCLUSIONS: Myomyous junctions form a complex muscle fiber network, which is thought to synchronize the activity of the CT muscle fibers. This is the first three-dimensional demonstration of the muscle fiber network in the CT muscle; this network may play a major role in the functional specificity of the CT muscle. PMID- 17415129 TI - Use of recombinant factor VIIa during excision of vascular anomalies. AB - BACKGROUND: The surgical excision of vascular anomalies is often accompanied with significant perioperative bleeding. Novel hemostatic agents, including recombinant factor VIIa (rVIIa), have been shown to reduce bleeding in hemophilia and trauma patients along with decreasing blood loss during various surgical procedures. The role of rVIIa during excision of vascular anomalies has not been examined. METHODS: A retrospective chart review of patients from 2001 to 2003 who received perioperative rVIIa during excision of vascular anomalies at one institution. RESULTS: Nine patients were identified who received perioperative rVIIa during removal of their vascular anomalies (7 venous malformations, 1 lymphatic-venous malformation, 1 arteriovenous malformation). All patients received at least one dose of rVIIa during the perioperative period (2/9 received 2 doses). An accurate account of hourly blood loss was reported, and could be graphed, in three patients. These patients displayed significant reductions in hourly blood loss after the administration of rVIIa. These cases are detailed in this report. Surgeons reported subjective improvements in operative bleeding, efficiency, and operative time in six of six patients after the administration of rVIIa intraoperatively. Reduced postoperative bleeding was reported in two patients who received rVIIa for persistent drain output. CONCLUSION: rVIIa may be an effective adjunct in improving the surgical efficiency and outcome of excised vascular anomalies. PMID- 17415130 TI - Outcome measurement for patients receiving hearing aid services. AB - OBJECTIVE: The purpose of this paper is to describe a useful patient report outcome measure for a hearing aid program that can be completed annually, with limited staff resources, and can be used to document performance excellence. STUDY DESIGN AND METHODS: The International Outcome Inventory for Hearing Aids (IOI-HA) questionnaire was mailed to all patients receiving new hearing aids and rehabilitative services during a 1 year period at Mayo Clinic (Rochester, MN). The data were analyzed to assess outcomes. RESULTS: Overall, patients reported a high level of satisfaction and an improved quality of life. The results compared favorably with published norms and with the previous year's results. CONCLUSION: The IOI-HA was implemented easily with little incremental cost. An annual project such as the one described can be used to demonstrate to administrators, patients, and payers that the clinic's rehabilitative hearing aid services have positive outcomes and that program quality is being monitored. PMID- 17415131 TI - An alternate approach for surgical dressing of bone-anchored hearing aid abutment sites. AB - OBJECTIVE: To present an alternative surgical dressing for bone-anchored hearing aid (BAHA) abutment sites, comparing it with the manufacturer's recommended "healing cap" in terms of split thickness skin graft (STSG) survival. STUDY DESIGN AND SETTING: A retrospective review of 30 patients who underwent unilateral BAHA implantation at a tertiary referral center. Patients were divided into two groups on the basis of the surgical dressing for the BAHA abutment site. Group 1 had a "healing cap" dressing, and group 2 had a bolster dressing. STSG survival was evaluated. RESULTS: Between May 2002 and July 2006, 30 patients underwent BAHA implantation. Seven patients received the "healing cap" dressing, and 23 patients received the bolster dressing. There was 100% STSG survival in the bolster dressing group and 71% skin graft survival in the healing cap group (P = .048). CONCLUSION: A traditional bolster dressing had improved STSG survival as compared with the manufacturer's recommended "healing cap." Additional benefits of a bolster are that it is maintenance free, well suited for noncompliant patients, and inexpensive. PMID- 17415132 TI - Evaluation of radiofrequency thermal ablation results in inferior turbinate hypertrophies by magnetic resonance imaging. AB - OBJECTIVES: Nasal obstruction caused by inferior turbinate hypertrophies is one of the most frequent problems in otolaryngology. Treatment of this disease may involve medical and surgical methods. Thermal ablation with radiofrequency energy has become quite popular in the recent years as one of the surgical methods used when medical therapy is not adequate. STUDY DESIGN: Prospective, clinical trial. METHODS: Thermal ablation with radiofrequency energy was used in this study on the inferior turbinates of 21 patients who had nasal obstruction caused by inferior turbinate hypertrophy only. The results were evaluated subjectively by preoperative and postoperative patient- and physician-assigned visual analogue scales and objectively by magnetic resonance imaging (MRI) investigation. RESULTS: By the end of the postoperative week 10, 64.76% recovery was detected according to the patient evaluation, and 40.75% recovery was detected according to the physician evaluation. Measurement of the average volumes of the inferior turbinates by MRI revealed a 8.70% postoperative reduction. The most significant change was detected in the anterior-posterior length measurement performed in the axial plane. CONCLUSIONS: These results suggest that thermal ablation with radiofrequency energy is an easily applied, efficient, and reliable technique in treatment of the inferior turbinate hypertrophy, and that anterior-posterior length measurement in the axial section of the inferior turbinate by MRI, which is thought as an objective evaluation method, could be an efficient diagnostic tool in detecting the efficiency of radiofrequency on inferior turbinate. PMID- 17415133 TI - Congenital cholesteatoma: risk factors for residual disease and retraction pockets--a report on 117 cases. AB - OBJECTIVES: To define predictors of residuals and retraction pockets (RP) in children operated on for congenital cholesteatoma (CC). DESIGN AND SETTING: Retrospective review (1996-2005), academic center. PATIENTS: One hundred seventeen patients treated for CC corresponding to modified Derlacki's criteria were included (median age, 6.5 yr). No case of RP at time of diagnosis, with a mean follow-up of 2.5 years after last surgery. MAIN OUTCOME MEASURES: Clinical and surgical data influencing outcome. Multivariate analysis. RESULTS: Two groups were defined after CC removal: group I (12 cases), no second look required and no case of subsequent re-intervention; group II (105 cases), planned second look always performed (mean delay, 12.1 mo), no difference of sex ratio (M/F = 2). Group I patients were younger than in group II (3.3 vs. 5.9 yr, P < .001). All of them had a normal contralateral eardrum and a disclosure of CC by routine examination (vs. 19% in group II, P < .001). In group I, the mass occupied one or two anterior quadrants (41.6% and 58.4%, respectively) versus more than two quadrants in 46.6% in group II. Residuals and RP rates were 41% and 15%, respectively (only in group II). Predictors for residuals were atticotomy (odds ratio [OR] 2.9, 95% confidence interval [CI] 1.3-6.7) and destruction of stapes (OR 4.3, 95% CI 1.7-10.5). Predictors for RP were eustachian tube extension (OR 6.8, 95% CI 1.7-26.8) and nonreconstructed atticotomy (OR 5.9, 95% CI 1.1-30.9). CONCLUSIONS: Young children with small CC had no recurrences. Residuals were more frequent in case of atticotomy and stapes destruction. RP occurred especially in cases of eustachian tube extension and if cartilage tympanoplasty was not performed. Tympanic and canal wall reinforcement should be considered in extensive CC. PMID- 17415134 TI - Clinical outcome after surgery for 75 patients with parotid sialadenitis. AB - OBJECTIVE: To analyze the clinical outcome of patients treated surgically for chronic parotid sialadenitis. STUDY DESIGN: A retrospective cohort study of patients whose clinicopathologic information had been collected prospectively onto a dedicated head and neck database. METHODS: Between 1987 and 2006, a total of 75 patients had 78 parotidectomies, 17 superficial and 61 near-total, to treat chronic parotid sialadenitis. The clinicopathologic data including treatment morbidity and outcome were analyzed. RESULTS: Temporary postoperative facial weakness occurred in 26 (33%) patients, and this was permanent, although partial, in one (1%) patient. There was no significant difference (P > .05) in the incidence of temporary facial nerve neuropraxia in the superficial (35%) and near total parotidectomy (33%) groups. Recurrence of symptoms was noted in two patients who had undergone near-total parotidectomy and none of the patients who had undergone superficial parotidectomy. Chronic parotid sialadenitis was effectively treated in 97% of patients undergoing parotidectomy. CONCLUSION: Near total parotidectomy is a safe and efficacious surgical treatment in the management of patients with debilitating severe chronic parotid sialadenitis. PMID- 17415135 TI - Use of computational modeling to predict responses to upper airway surgery in obstructive sleep apnea. AB - OBJECTIVES: Despite the well-recognized consequences of obstructive sleep apnea (OSA), its treatment remains unsatisfactory. Therapeutic strategies are complicated by often poor adherence in the case of continuous positive airway pressure or the highly variable efficacy in the case of many upper airway surgeries. Computational models of the upper airway using finite element analysis to simulate the effects of various anatomic and physiologic manipulations on pharyngeal mechanics could be helpful in predicting surgical success. STUDY DESIGN: Computational and physiologic study. METHODS: Using representative OSA magnetic resonance images and experimentally measured upper airway dilator muscle activities, we developed a working two-dimensional and a partial three dimensional model of the upper airway. RESULTS: As predicted from experimental measurements, the OSA model airway has a closing pressure of -2 cm H2O. Manipulations such as palatal stiffening, palatal resection, and tongue stiffening all have demonstrable effects on pharyngeal mechanics. We have also developed a partial three-dimensional OSA model in which we simulate the mechanics of the pharyngeal airway in the mid-sagittal and parasagittal slices, spanning more than 1 inch in thickness. Using this model, we have observed important effects of tongue and palatal stiffening on anteroposterior collapse of the pharyngeal airway. CONCLUSIONS: Our data suggest that computational modeling is feasible and can be used to generate hypotheses for subsequent clinical trials regarding anatomic manipulations in OSA. We further believe that the goal of individualizing OSA therapy on the basis of underlying mechanisms could be facilitated by computational modeling. PMID- 17415136 TI - Tonsil surgery in youths: good results with a less invasive method. AB - OBJECTIVE: Comparison of two types of tonsil surgery for 16- to 25-year-old patients, with respect to primary morbidity, snoring, and recurrent infections after 1 year. Teenagers and young adults are a significant proportion (26%) of the population that receive tonsil surgery each year and appear to suffer more pain than younger children. Recurrent tonsillitis, in combination with obstructive problems, is the main indication for surgery. METHOD: One hundred fourteen patients 16 to 25 years of age were randomized to tonsillotomy (TT) with radiosurgery (RF) (Ellman International) or to cold tonsillectomy (TE). Pain and analgesics were logged until patients were pain free. RESULTS: Thirty-two patients were operated on with TT and 44 with TE. The TT group had less blood loss during surgery and no postoperative bleedings, compared with the TE group (2 primary and 4 late hemorrhages). The TT group recorded significantly less pain from the first day, had less need of analgesics (diclofenac and paracetamol), and were pain free and in school/at work 4 days earlier than the TE group. After 7 days, the TE patients had lost a mean of 1.8 kg compared with TT, with no significant weight loss. After 1 year, both groups were satisfied. The positive effect on snoring was the same for both groups. There were few throat infections in both groups. CONCLUSION: TT with RF is an effective method for tonsil surgery for many teenagers and young adults, with much less postoperative morbidity than regular TE. Long-term follow-up is necessary. PMID- 17415137 TI - Rethinking nasal osteotomies: an anatomic approach. AB - OBJECTIVES: To review our experience with our anatomic (medial, transverse, and lateral) nasal osteotomy technique for correcting the asymmetric bony nasal vault in esthetic and functional rhinoplasty. METHODS: All patients undergoing anatomic nasal osteotomy technique by the senior author (y.d.) from August 1997 to August 2005 with a minimum follow-up of 6 months were reviewed. Preoperative and postoperative photographs and clinical examination were analyzed to determine restoration of a symmetric bony nasal vault configuration. RESULTS: A total of 322 patients met the inclusion criteria. Percent of patients who had favorable results with restoration of a symmetric bony nasal vault configuration was 98.1%. Six (1.9%) patients required revision surgery at the level of the nasal bones with repeat osteotomies or dorsal rasping. All six of these patients had favorable outcomes after revision surgery. CONCLUSIONS: The anatomic nasal osteotomy technique represents an effective, simple, and logical approach for correcting asymmetric bony nasal vault. PMID- 17415138 TI - Biofilms in ear, nose, and throat infections: how important are they? AB - BACKGROUND: Biofilms present a new challenging concept in sustaining chronic, common antibiotic-resistant ear, nose, and throat (ENT) infections. They are communities of sessile bacteria embedded in a matrix of extracellular polymeric substances of their own synthesis that adhere to a foreign body or a mucosal surface with impaired host defense. The aim of this paper is to review the literature on ENT diseases that can be attributed to biofilm formation and to discuss options for future treatment. MATERIALS AND METHODS: Literature review from Medline and database sources. Electronic links and related books were also included. STUDY SELECTION: Controlled clinical trials, animal models, ex vivo models, laboratory studies, retrospective studies, and systematic reviews. DATA SYNTHESIS: Biofilm formation is a dynamic five-step process guided by interbacterial communicating systems. Bacteria in biofilms express different genes and have markedly different phenotypes from their planktonic counterparts. Detachment of cells, production of endotoxin, increased resistance to the host immune system, and provision of a niche for the generation of resistant organisms are biofilm processes that could initiate the infection process. Effective prevention and management strategies include interruption of quorum sensing, inhibition of related genes, disruption of the protective extrapolymer matrix, macrolides (clarithromycin and erythromycin), and mechanical debridement of the biofilm-bearing tissues. With regard to medical indwelling devices, surface treatment of fluoroplastic grommets and redesign of cochlear implants could minimize initial microbial colonization. CONCLUSION: As the role of biofilms in human infection becomes better defined, ENT surgeons should be prepared to deal with their unique and tenacious nature. PMID- 17415139 TI - The treatment of idiopathic sudden sensorineural hearing loss using pulse therapy: a prospective, randomized, double-blind clinical trial. AB - OBJECTIVES: The etiology and treatment of idiopathic sudden sensorineural hearing loss (ISSHL) is still unclear. The anti-inflammatory effect of corticosteroids is thought to play an important part in the recovery from ISSHL. We aimed to determine whether a more powerful anti-inflammatory technique using pulse therapy is effective in the treatment of ISSHL. METHODS: In a randomized, prospective, double-blind, multicenter clinical trial, we recruited 81 patients with ISSHL. Patients were randomly allocated to pulse therapy (300 mg dexamethasone for 3 consecutive days followed by 4 days of placebo) or control treatment (prednisone 70 mg per day tapered in steps of 10 mg per day to 0 mg). The primary outcome was hearing recovery as measured by pure-tone audiometry and speech audiometry after 12 months. Secondary outcomes were subjective parameters such as hearing recovery, tinnitus, vertigo, and a pressure sensation in the ear. RESULTS: The overall improvement in pure-tone thresholds and speech discrimination scores was not significantly better in patients who were given dexamethasone than those who were given standard prednisone. Hearing improved from 71 dB HL to 36 dB HL in the dexamethasone group and from 75 dB HL to 42 dB HL in the prednisone group. Speech discrimination scores of 100% were achieved by 64% of dexamethasone-treated patients and by 57% of the prednisone group. CONCLUSION: Pulse therapy is equally effective and safe as standard-dose prednisone. Pulse therapy suppresses both humoral and cellular immune responses and therefore has a wider anti-inflammatory effect. PMID- 17415140 TI - Clinical symptomatology and paranasal sinus involvement with nasal septal perforation. AB - OBJECTIVE: Determine the symptom manifestations, clinical impact, and incidence of chronic rhinosinusitis (CRS) in patients with newly diagnosed nasal septal perforation. METHODS: A consecutive series of adult patients with nasal septal perforation were prospectively studied at the time of endoscopic diagnosis with the rhinosinusitis symptom inventory (RSI) and sinus computed tomography (CT). Patients' symptoms in the RSI symptom domains were computed. From the CT scan, septal perforation size and Lund scores were obtained. A separate (control) cohort of patients with CRS without septal perforation was matched to these patients for age, sex, and Lund score. RSI symptom domain comparisons were conducted between groups to determine the additional symptom burden conferred by septal perforation. RESULTS: Thirty-three patients with septal perforation were enrolled (mean age, 48.2 yr; 69.7% female). Mean perforation size was 1.9 (SD, 2.1) cm. The mean Lund score was 5.8 (SD, 5.3); 16 (57.1%) patients met radiographic criteria for a concurrent diagnoses of CRS. Patients with septal perforation reported significant nasal and facial symptom domain scores (56.8 and 47.0, respectively [range, 0-100]). Oropharyngeal and systemic symptoms were less severe (29.7 and 34.7, respectively). However, after comparison with the matched control patients, no statistically significant differences were identified in sinonasal symptoms between patients with and without septal perforation (all P > .131). CONCLUSIONS: Concurrent CRS may frequently accompany nasal septal perforation and may require appropriate treatment along with the perforation itself. The presence of septal perforation does not appear to significantly augment symptom severity in CRS. PMID- 17415141 TI - Expressions of aquaporin-2, vasopressin type 2 receptor, transient receptor potential channel vanilloid (TRPV)1, and TRPV4 in the human endolymphatic sac. AB - OBJECTIVE: To localize aquaporin (AQP)2, vasopressin type 2 receptor (V2-R), and transient receptor potential channel vanilloid subfamily 1, 4 (TRPV1, TRPV4) in the human endolymphatic sac (ES). METHODS: Three samples of human ES were sampled during the removal of vestibular schwannoma by way of the translabyrinthine approach. The samples were immediately fixed in 4% paraformaldehyde and embedded in OCT compound; immunohistochemistry was performed with AQP2, V2-R, TRPV1, and TRPV4 polyclonal antibodies. RESULTS: AQP2, V2-R, TRPV1, and TRPV4 proteins were detected in the epithelial layer of the ES but were not observed in connective tissue around the ES. TRPV1 was also expressed in blood vascular endothelial cells of the connective tissue of ES. CONCLUSIONS: AQP2, V2-R, and TRPV4 were expressed in the luminal epithelium of human ES. The same characteristic distribution of water and ion channels is seen in the kidney, where a significant amount of fluid is filtrated and resorbed. ES probably plays an active role in the homeostasis of the endolymph. PMID- 17415142 TI - Animal model for cholesteatoma induced in the gerbil: will the profiles of differentiation/growth-regulatory markers be similar to the clinical situation? AB - INTRODUCTION: Cholesteatoma is a benign tumor of the middle ear characterized by an aggressive and invasive potential. The only current treatment being surgery, it is important to have access to a reliable animal model to study and better understand cholesteatoma pathogenesis. Our study aimed to examine the biological validity of the most common experimental model of cholesteatoma: the Mongolian gerbil. MATERIAL AND METHODS: We have induced cholesteatoma by surgical ligature of the gerbil's external auditory duct. Quantitative comparison of eight biological markers involved in inflammation (macrophage migration inhibitory factor [MIF]), cell differentiation (retinoic acid receptors-alpha, -beta, and gamma), and cell adhesion/apoptosis (galectins-1, -3, -7, and -8). The immunohistochemical staining was quantified by computer-assisted microscopy. RESULTS: Two immunohistochemical parameters were determined in sections. The labeling index (LI) represents the percentage of tissue area specifically stained, and the mean optical density (MOD) denotes the staining intensity index. The LI reveals statistically significant differences for each marker tested. The MOD also shows statistically significant differences except for MIF (P = .259). CONCLUSION: From the panel of markers, the majority of staining parameters was statistically significantly different between sections of the animal model and clinical specimen. These data do not support the concept of complete validity of the popular animal model. PMID- 17415143 TI - Demands on caring relatives of head and neck cancer patients. AB - BACKGROUND: Relatives of cancer patients experience high levels of stress that influence the quality of life of these individuals. To investigate whether there is a necessity for simultaneous supportive care of patient relatives, we performed for the first time a study asking the closest relatives of head and neck cancer patients about their needs during and after the treatment to consider how to optimize the situation for such patient groups. MATERIAL AND METHODS: Patients' relatives were assessed using an anonymous self-report questionnaire that was established in our department by expanding on a questionnaire for cancer patients' relatives from the psycho-oncologic society in Switzerland. The evaluation was multidimensional, cancer specific, and relative based. RESULTS: Relatives feel confronted themselves with cancer, although indirectly. The majority of the respondents were of the opinion that simultaneous psychological care of the patients and for the caring relatives would be helpful to cope with the situation. CONCLUSION: This study shows the significant impact of cancer on caring relatives of head and neck cancer patients. In our opinion, health services should become more aware of this potential to ensure that the needs of the involved patient relatives are met as well as those of the patients. PMID- 17415144 TI - Key messages from the National Prospective Tonsillectomy Audit. AB - OBJECTIVES: Investigation of the occurrence of postoperative hemorrhage after tonsillectomy and risk factors for these complications. STUDY DESIGN: Prospective National Audit with electronic web-based data collection from the National Health Service and independent hospitals in England and Northern Ireland. METHODS: Consenting patients undergoing tonsillectomy between July 2003 and September 2004 were included. A central database of patient and surgical variables was developed for analysis of primary and secondary postoperative hemorrhage and development of risk models for tonsillectomy complications. RESULTS: The Audit received data from 277 hospitals in England and Northern Ireland on 40,514 patients. Analysis was conducted on 33,921 (84%) who gave consent. Over the whole Audit 1,197 (3.5%) postoperative hemorrhages were recorded. One hundred eighty-eight (0.6%) patients sustained a primary hemorrhage and 1,033 (3%) a secondary hemorrhage (24 had both). Elevated hemorrhage rates were observed in tonsillectomies performed using diathermy for dissection and hemostasis compared with cold steel dissection and ties for hemostasis. National guidance issued midway through the Audit influenced practice and reduced hemorrhage rates. Risk models for postoperative hemorrhage were developed incorporating the effect of the guidance. For hemorrhage, compared with the cold steel group, bipolar diathermy tonsillectomy had an odds ratio of 2.47 (1.81-3.36), P < .0001, and bipolar diathermy scissors tonsillectomy an odds ratio of 3.20 (2.09-4.90), P < .0001. Use of bipolar diathermy for hemostasis only after cold steel dissection carried an intermediate risk, odds ratio 1.57 (1.16-2.13), P = .004. CONCLUSIONS: The results confirm that "hot" tonsillectomy techniques carry a substantially elevated risk of postoperative hemorrhage when diathermy is used as a dissection tool in tonsillectomy. PMID- 17415145 TI - Optimal graft thickness for different sizes of tympanic membrane perforation in cartilage myringoplasty: a finite element analysis. AB - OBJECTIVE: The purpose of this study was to determine, using finite element analysis, the optimal graft thickness for cartilage myringoplasty in patients with different sizes of tympanic membrane (TM) perforations. STUDY DESIGN: We developed a cartilage plate-TM-coupled model using high-resolution computed tomography and finite element analysis. The geometric models of the perforated TM were generated using Patran and ANSYS software. METHOD: Three different sizes of TM perforations (15%, 55%, and 85%, representing small, medium, and large perforations, respectively) were created in the pars tensa. A cartilage plate was used to repair the eardrum perforation, and the new TM-cartilage coupled complex was loaded into our three-dimensional biomechanical model for analysis. The frequency-amplitude responses for different cartilage thicknesses were compared with those for natural TM. RESULTS: Our results show that, first, in cases with 85% perforation, the frequency-amplitude responses that were most similar to natural TM at lower frequencies were for graft thicknesses of 0.2 mm and for 0.1 mm at higher frequencies. Second, in cases with 55% posterior perforation of the TM, assessment of the predicted vibration amplitude of different thicknesses of the cartilage plate showed that a cartilage plate of less than 0.2 mm had a frequency response function similar to that of a natural TM in umbo and stapes footplate displacement. Finally, for a central perforation involving 15% of the TM, a cartilage plate of less than 1.0 mm showed a frequency response function similar to that of TM in umbo and stapes-footplate displacement. CONCLUSIONS: On the basis of our biomechanical analysis, the optimal thickness of a cartilage graft for myringoplasty appears to be 0.1 to 0.2 mm for medium and large TM perforations. For small perforations, a cartilage of less than 1.0 mm is a good compromise between mechanical stability and low acoustic transfer loss. PMID- 17415146 TI - Plasma radiofrequency preceded by pressure recording enhances success for treating sleep-related breathing disorders. AB - OBJECTIVES: Sleep-related breathing disorders (SRBD) that cannot be treated conservatively are commonly treated using uvulopalatopharyngoplasty, although success rates are generally less than 70%. The purpose of this study was to assess a plasma-mediated radiofrequency (RF)-based coblation assisted upper airway procedure (CAUP) to treat SRBD patients determined to have obstruction localized to the upper (mid)-pharyngeal region. STUDY DESIGN: Prospective case series (n = 40). METHODS: Six patients had socially bothersome snoring, and 34 patients had mild to moderate sleep apnea (apnea/hypopnea index [AHI] <20), with the primary level of obstruction (>50%) within the upper pharyngeal region as determined using whole night recordings, including airway pressure fluctuation monitoring (ApneaGraph, MRA-Medical Ltd, Gloucestershire, UK). CAUP consisted of making a lateral palatal incision, ablating three upward channels on each side of the midline (fan-shaped) into the soft palate using a plasma mediated RF-based device (ArthroCare Corporation, Austin, TX), and performing a partial uvulectomy. Clinical outcomes included the Epworth Sleepiness Scale (ESS), partner rating of snoring using a visual analogue scale (VAS), and night-time apnea and hypopnea events (AHI, hypopnea index [HI], apnea index [AI]). RESULTS: Patients were 28 to 68 (46 +/- 12) years old; 28 (70%) were male. Preoperatively, clinical assessment scores (median +/- interquartile range) were as follows: ESS (11.0 +/- 3.0), VAS (8.15 +/- 1.00), AHI (9.58 +/- 5.58), HI (9.00 +/- 5.29), AI (0.333 +/- 0.625). After CAUP, no postoperative scarring, fibrosis, or any other clinically significant side effects were observed. Postoperatively (9.1 +/- 1.5; 7-15 mo), ESS (4.0 +/- 1.0), VAS (2.70 +/- 1.38), AHI (3.75 +/- 2.92), HI (3.58 +/- 2.50), and AI (0.167 +/- 0.167) were significantly improved (P < .001). CONCLUSION: CAUP preceded by site-specific obstruction diagnosis using pressure recording is a well-tolerated outpatient treatment that is well suited for treating SRBD. PMID- 17415147 TI - Olfaction and olfactory epithelium in mice treated with zinc gluconate. AB - OBJECTIVE: We assessed whether a nasal spray containing zinc gluconate (ZG) compromises the integrity of olfactory epithelium and olfactory function. METHODS: Axonal transport of horseradish peroxidase from olfactory epithelium to the olfactory bulb was studied in 2- to 21-day survival mice given intranasal injections of 2, 8, or 50 microL of ZG (approximately 4, 15, and 94 times the equivalent recommended human dose). Other similarly treated mice were tested using precision olfactometry to detect and discriminate odors. RESULTS: Anatomic changes were graded as a function of dose and survival time. Two microliter injections had no discernable effect. while the 50 microL volume produced substantial deafferentation of input to the olfactory bulb in short-survival cases. Nearly complete restitution of input occurred within 3 weeks. At each volume and survival time, zinc sulfate (ZS) had a greater effect. Behaviorally, 2 microL and 8 microL ZG-treated mice and those given multiple injections of 2 microL ZG performed as well as controls, whereas those given 50 microL were hyposmic but not anosmic. ZS-treated mice performed more poorly, and those injected with 50 microL were anosmic for the first 8 to 10 test days. CONCLUSIONS: A massive dose of a ZG nasal spray did cause a transient disruption of the olfactory epithelium and compromised olfaction. More moderate volumes, even those far in excess of a recommended dose, were largely without effect on odor detection and discrimination tasks. These outcomes fail to support the claims from recent clinical case reports that use of a ZG-containing nasal spray can produce anosmia. PMID- 17415148 TI - Vascularized mucoperiosteal pull through flap for closure of large septal perforation: a new technique. PMID- 17415149 TI - Management of thyroglossal duct cysts after failed Sistrunk procedure. PMID- 17415150 TI - Using stereolithographic models to plan mandibular reconstruction for advanced oral cavity cancer. PMID- 17415151 TI - Endoscopic endonasal resection of a synchronous pituitary adenoma and a tuberculum sellae meningioma: technical case report. AB - OBJECTIVE: The presence of a sellar macroadenoma with a concomitant tuberculum sellae meningioma typically requires a craniotomy for extirpation of the meningioma. We describe a single endoscopic approach for resection of both lesions. PRESENTATION: A 52-year-old woman with complaints of continuous headache and right temporal visual field loss was found to have a 1.9 x 2.1 x 1.7-cm enhancing sellar mass on magnetic resonance imaging scans associated with a second enhancing extra-axial lesion at the planum sphenoidale measuring 1.0 x 0.6 cm and encroaching on the right optic nerve. INTERVENTION: After the endoscopic transnasal resection of the pituitary tumor, the planum sphenoidale was drilled and the underlying dura was incised. The suprasellar tumor was identified and completely resected. Histological evaluation confirmed the concomitant presence of a meningioma and pituitary adenoma. CONCLUSION: With advances in endoscopic and image-guidance technology and increasing understanding of the endoscopic anatomy of the sellar region, surgeons are capable of reaching both intrasellar and suprasellar/anterior cranial base region tumors through a single endoscopic approach. The use of endoscopes in transsphenoidal approaches may obviate the need for additional craniotomies in properly selected patients. PMID- 17415154 TI - Functional brain mapping and its applications to neurosurgery. AB - Functional brain mapping may be useful for both preoperative planning and intraoperative neurosurgical decision making. "Gold standard" functional studies such as direct electrical stimulation and recording are complemented by newer, less invasive techniques such as functional magnetic resonance imaging. Less invasive techniques allow more areas of the brain to be mapped in more subjects (including healthy subjects) more often (including pre- and postoperatively). Expansion of the armamentarium of tools allows convergent evidence from multiple brain mapping techniques to bear on pre- and intraoperative decision making. Functional imaging techniques are used to map motor, sensory, language, and memory areas in neurosurgical patients with conditions as diverse as brain tumors, vascular lesions, and epilepsy. In the future, coregistration of high resolution anatomic and physiological data from multiple complementary sources will be used to plan more neurosurgical procedures, including minimally invasive procedures. Along the way, new insights on fundamental processes such as the biology of tumors and brain plasticity are likely to be revealed. PMID- 17415155 TI - Percutaneous transforaminal lumbar interbody fusion for the treatment of degenerative lumbar instability. AB - OBJECTIVE: Percutaneous spinal instrumentation techniques may be helpful to reduce approach-related morbidity inherent to conventional open surgery. This article reports technique, clinical outcomes, and fusion rates of percutaneous transforaminal lumbar interbody fixation (pTLIF). Results are compared with those of mini-open transforaminal lumbar interbody fixation (oTLIF) using a muscle splitting (Wiltse) approach. METHODS: pTLIF was performed in 43 patients with single-level and 10 patients with bi- or multilevel lumbar discopathy or degenerative pseudolisthesis resulting in axial back pain and claudication, pseudoradicular, or radicular symptoms. Decompression, discectomy, and interbody cage insertion were performed through 18-mm tubular retractors followed by percutaneous pedicle screw-rod fixation. Clinical outcome was assessed by early postoperative pain scores (visual analog score) and standardized functional outcome questionnaires (American Academy of Orthopedic Surgeons lumbar spine and Roland-Morris low back pain score). Fusion rates were assessed by thin-slice computed tomographic scan at 16 months. Clinical outcome, time in the operating room, intraoperative blood loss, and postoperative access-site pain were compared with an institutional reference series of 67 oTLIF procedures. RESULTS: Excellent and good clinical results were obtained in 46 (87%) out of 53 patients at 16 months. The time spent in the operating room was equivalent and the blood loss reduced compared with oTLIF (P < 0.01). There was no morbidity related to instrumentation. Postoperative pain was significantly lower after pTLIF after the second postoperative day (P < 0.01). The overall clinical outcome was not different from oTLIF at 8 and 16 months. CONCLUSION: pTLIF allows for safe and efficient minimally invasive treatment of single and multilevel degenerative lumbar instability with good clinical results. Further prospective studies investigating long-term functional results are required to assess the definitive merits of percutaneous instrumentation of the lumbar spine. PMID- 17415156 TI - Microsurgical technique and results of a series of 63 spheno-orbital meningiomas. AB - OBJECTIVE: For this study, spheno-orbital meningiomas (SOMs) are defined as intraosseus meningiomas at the base of the anterior and middle cranial fossa, involving the sphenoid wing and orbit associated with a carpet-like, soft tissue component. We describe a surgical series of 63 SOMs, including surgical technique, complications, and recurrences. METHODS: For this retrospective series, patient charts of those who underwent operation for SOM by or under the supervision of the senior author (JS) at two institutions were identified and reviewed. Follow-up data was collected for up to 17 years (median, 4.5 yr). RESULTS: Between 1983 and 2003, 63 patients with a mean age of 51 years underwent operation for SOM. The most common preoperative sign was proptosis (79%), followed by visual acuity deficits (27%). Preoperative symptoms included a visual acuity below 0.8 (47%) and visual field deficits of 32%. Bony resection included the lesser sphenoid wing, the lateral orbital wall, the orbital roof, the optic canal, and the middle fossa base. Periorbital tumor infiltration led to intraorbital resection in 32 cases, and five cases presented a solid intraorbital tumor. After surgery, proptosis improved in 77% of the patients; 64% of the patients had improved visual acuity, and 58% of the patients with preoperative deficits had improved visual fields. Postoperatively, 12 patients showed temporary oculomotor deficits, and eight patients showed permanent oculomotor deficits. Temporary (n = 9) and persisting (n = 2) ptosis was observed. The median follow-up period was 4.5 years. Seventy-six percent of patients had tumor residuals, of which 61% were stable and 39% were progressive. Eleven tumor residuals were operated and four were treated by radiation. CONCLUSION: Complete surgical resection of SOMs is frequently impossible because the involvement of delicate structures of the orbital cone is common. Although some persisting neurological deficits are possible, proptosis and other visual deficits are often relieved. Two-thirds of tumor rests remained stable during the follow-up period. Consequently, the surgical aim should be the relief of leading symptoms rather than radical resection. PMID- 17415157 TI - Thoracic kyphotic deformity reduction with a distractible titanium cage via an entirely posterior approach. AB - OBJECTIVE: Surgical correction of thoracic kyphotic deformity is often associated with significant surgical and neurological morbidity and unsatisfactory reduction of kyphosis, especially in patients who cannot tolerate anterior thoracic procedures because of associated comorbidity. We describe a technique in which kyphotic deformity of the thoracic and thoracolumbar spine is corrected, decompressed, and stabilized with a circumferential fixation construct from a lone posterior approach. METHODS: We reviewed the radiographic and clinical outcomes of seven patients undergoing vertebrectomy via a bilateral modified costotransversectomy approach followed by posterior placement of a distractible cage, reduction of the deformity via cage distraction, and supplemental dorsal instrumentation. All patients possessed thoracic/thoracolumbar kyphosis; however, a transthoracic approach was thought to be high risk because of medical comorbidity. RESULTS: Seven patients underwent this procedure for thoracolumbar kyphosis resulting from a spinal tumor, osteomyelitis, and fracture. Vertebrectomies were performed at T2-T3, T4-T5, T5-T6, T12-L1, and L1. The mean preoperative kyphosis was 28.6 degrees, the mean postoperative kyphosis at the time of the final follow-up examination was 12.1 degrees, and the mean change in kyphosis was 53%. The mean long-term follow-up period was approximately 16 months. At the time of the final follow-up examination for all patients, there was no decline in neurological function, and pain management consisted of minimal use of oral narcotics. CONCLUSION: This technique allows for circumferential decompression of the spinal cord via a posterior approach in patients with thoracic kyphotic deformities who cannot tolerate anterior thoracic approaches. In addition, in situ distraction of the expandable cage allows correction of sagittal imbalance and restores height without the potential loss of spinal height associated with osteotomies. PMID- 17415158 TI - Posterior keyhole corpectomy with percutaneous pedicle screw stabilization in the surgical management of lumbar burst fractures. AB - OBJECTIVE: The authors present a new method of minimally invasive surgical management of lumbar burst fractures through the posterior approach. The method includes minimally invasive corpectomy and interbody fusion, both of which are performed through a keyhole approach, and percutaneous pedicle screw fixation of the fracture. The technique of the posterior keyhole corpectomy presented in this report is a novel and original concept of the first author (AM). The percutaneous pedicle screw stabilization is performed with the use of a percutaneous instrumentation system (Sextant; Medtronic, Inc., Minneapolis, MN). The Sextant system has been dedicated and used in nontrauma degenerative cases; the novel aspect of this system is its application in spine fractures. Indications for the method include Denis classification subtype B or Magerl subtype A.3.1 burst fractures. Both subtypes represent fractures with failure and retropulsion of the upper part of the vertebral body. METHODS: The clinical experience of this study includes four cases of burst fractures with significant retropulsion and occlusion of the spinal canal. Long-term results were assessed at a minimum follow-up period of 1 year (maximum, 3.5 yr). The follow-up assessments included: 1) the quality of decompression and reconstruction of the spinal canal (computed tomographic and magnetic resonance imaging scanning); 2) the stability of the operated segment (dynamic x-rays); 3) the quality of interbody fusion (computed tomographic scanning and dynamic x-rays); and 4) correction of the fracture kyphosis and its postoperative loss (measurements of Cobb angles for the assessment of sagittal plane deformity). The minimum armamentarium requirements for this method include a typical micro lumbar discectomy retractor set; a surgical microscope; two-plane intraoperative fluoroscopy; and a system for percutaneous pedicle screw stabilization (Sextant). "Posterior keyhole corpectomy" indicates corpectomy of the posterior upper half of the vertebral body or removal of the retropulsed bone fragment via two keyhole skin incisions on both sides of the spinous process (each skin incision measures 2 to 3 cm long). Exposure of the retropulsed fragment (the posterior upper part of the vertebral body) is achieved by medial or complete facetectomy along with complete or medial resection of the pedicle. This has to be performed bilaterally. Percutaneous stabilization requires four additional stab skin incisions. RESULTS: We observed no surgery-related complications (neurological, hardware, dural tears, or deep or superficial wound infections); there was perfect decompression and clearance of the spinal canal (confirmed by computed tomographic and magnetic resonance imaging scanning); and there was solid stability at the affected segments (confirmed by dynamic x-rays). Healed fusion was noted in all patients but one. The latter patient had no clinical symptoms of spinal instability. Kyphotic deformity was corrected and reversed into lordosis in three patients. Loss of deformity correction was noted in all patients; however, all patients retained lordotic alignment of the affected segment. CONCLUSION: The advantages of this method include sparing the posterior elements (lamina, spinous process, supraspinous and interspinous ligaments, and paravertebral muscles), safety of the decompression provided by the use of a surgical microscope, and perfect illumination of the operating field. The drawbacks of the method include limitation to certain types of burst fractures, the method is surgically demanding, and the method requires development of a special retractor system to eliminate the cumbersome alternate insertion and the reinsertions of the typical microdiscectomy retractor set. PMID- 17415159 TI - Ventricular catheter placement with a frameless neuronavigational system: a 1 year experience. AB - OBJECTIVE: Accurate placement of ventricular catheters decreases the incidence of proximal catheter failure. The use of a frameless, interactive neuronavigational system can optimize catheter placement. METHODS: Thirty-four ventricular catheters were placed using a Medtronic electromagnetic frameless neuronavigational system (Medtronic Navigation, Inc., Louisville, CO) during a 12 month period. The patients ranged in age from 11 months to 79 years; the mean age was 40.8 years. Nineteen male and 12 female patients participated in the study. The indications for ventricular catheter placement included obstructive hydrocephalus, normal pressure hydrocephalus, pseudotumor cerebri, intrathecal therapy, and tumor cyst aspiration. RESULTS: No proximal failures have been reported to date. One infection necessitated shunt removal. Three postoperative deaths occurred because of non-catheter-related events. CONCLUSION: Frameless neuronavigation in the placement of ventricular catheters assures accurate catheter placement, thereby decreasing the incidence of proximal catheter failure. The absence of rigid head fixation allows additional cohorts to benefit from the apparatus. The use of the electromagnetic system provides a safe, simple, and easy adjunct to optimal catheter placement. PMID- 17415160 TI - Endoscopic transaqueductal removal of fourth ventricular neurocysticercosis with an angiographic catheter. AB - OBJECTIVE: Fourth ventricular neurocysticercosis (FVNCC) usually presents with obstructive hydrocephalus. Available treatment options are medical, external cerebrospinal fluid diversion, microsurgical, or endoscopic removal alone or in combination. We present our experience of transaqueductal removal of FVNCC by angiographic catheter with endoscopic third ventriculostomy with a rigid endoscope. METHODS: Ten patients (five male and five female patients; age range, 12-45 yr; mean, 23.2 years) with FVNCC with obstructive hydrocephalus underwent endoscopic removal along with endoscopic third ventriculostomy in a single sitting, through a frontal precoronal burr hole. Diagnosis was established on imaging and confirmed on histology in all of the cases. The Gaab Universal Endoscope System along with 4-mm, 30-degree rigid telescopes was used to enter the third ventricle, and a cut length of angiographic catheter was negotiated through the aqueduct for removal of FVNCC. RESULTS: Removal of the cyst was performed in all cases. A 30-degree rigid telescope provided excellent image quality, with the ability to address intra-FVNCC through the dilated aqueduct with a curved-tip catheter. None of these patients required any further surgery. There were no significant operative or postoperative complications in any of the cases. All of the patients were asymptomatic, with an average follow-up of 18 months. CONCLUSION: Transaqueductal removal of an intra-fourth ventricular cyst along with endoscopic third ventriculostomy with a rigid endoscope and catheter is an effective treatment and obviates the need for posterior cranial fossa exploration. PMID- 17415161 TI - In situ hinge craniectomy. AB - OBJECTIVE: To describe an optional method for performing decompressive craniectomy using in situ hinge craniectomy technique in patients with traumatic brain injury and stroke. METHODS: Sixteen patients underwent surgery for treatment of presenting pathology followed by the placement of hinge craniectomy. The technique is detailed. RESULTS: Six patients with traumatic head injury and 10 with stroke underwent treatment of their primary pathologies with subsequent hinge craniectomy. Of these patients, more than half underwent refixation of the hinge in a minor procedure after recovery. No patient had complications related to this technique and none required further cranial decompression. In patients with intracranial pressure monitoring, all displayed values in the normal range. CONCLUSION: In this limited study, in situ hinge craniectomy proved useful in the treatment of patients experiencing stroke or traumatic brain injury. This procedure has the potential to eliminate the additional second incision to explant the bone flap or the refrigeration storage of the bone flap. Also, the second operation to restore the cranial contour by reimplanting the bone flap or by the creation of a cranioplasty with artificial material would not be necessary. PMID- 17415162 TI - Electrocorticographic frequency alteration mapping: a clinical technique for mapping the motor cortex. AB - OBJECTIVE: Electrocortical stimulation (ECS) has been well established for delineating the eloquent cortex. However, ECS is still coarse and inefficient in delineating regions of the functional cortex and can be hampered by after discharges. Given these constraints, an adjunct approach to defining the motor cortex is the use of electrocorticographic signal changes associated with active regions of the cortex. The broad range of frequency oscillations are categorized into two main groups with respect to the sensorimotor cortex: low and high frequency bands. The low frequency bands tend to show a power reduction with cortical activation, whereas the high frequency bands show power increases. These power changes associated with the activated cortex could potentially provide a powerful tool in delineating areas of the motor cortex. We explore electrocorticographic signal alterations as they occur with activated regions of the motor cortex, as well as its potential in clinical brain mapping applications. METHODS: We evaluated seven patients who underwent invasive monitoring for seizure localization. Each patient had extraoperative ECS mapping to identify the motor cortex. All patients also performed overt hand and tongue motor tasks to identify associated frequency power changes in regard to location and degree of concordance with ECS results that localized either hand or tongue motor function. RESULTS: The low frequency bands had a high sensitivity (88.9 100%) and a lower specificity (79.0-82.6%) for identifying electrodes with either hand or tongue ECS motor responses. The high frequency bands had a lower sensitivity (72.7-88.9%) and a higher specificity (92.4-94.9%) in correlation with the same respective ECS positive electrodes. CONCLUSION: The concordance between stimulation and spectral power changes demonstrate the possible utility of electrocorticographic frequency alteration mapping as an adjunct method to improve the efficiency and resolution of identifying the motor cortex. PMID- 17415163 TI - The use of multiplanar trajectory planning in the stereotactic placement of depth electrodes. AB - OBJECTIVE: To assess the value of multiplanar reconstruction software in trajectory planning for depth electrode insertion in medically refractory epilepsy. METHODS: A series of 29 patients undergoing frame-based hippocampal depth electrode insertion were identified. In 19 patients, preoperative trajectory planning was conducted in axial, coronal, and sagittal planes using standard-axis software. In 10 patients, preoperative trajectory planning was conducted with multiplanar reconstruction software. Postoperative magnetic resonance imaging scans were evaluated to study the quality of insertion. Target accuracy was assessed by measuring the mean shortest distance to strictly defined hippocampal borders in the coronal plane ("coronal deviation"). Additionally, the number of electrode contacts placed within the amygdalohippocampal structure was assessed. RESULTS: With the use of multiplanar reconstruction software, there was a statistically insignificant increase in coronal deviation (standard-axis software group, 0.09 +/- 0.50 mm; multiplanar reconstruction group, 0.37 +/- 1.16 mm). However, the use of multiplanar planning strategies resulted in approximately one additional electrode contact inserted in the amygdalohippocampal structure (standard-axis software group, 3.42 +/- 0.89; multiplanar reconstruction group, 4.36 +/- 0.93; P < 0.01). CONCLUSION: The use of reconstructed planes in preoperative trajectory planning allows for the insertion of additional electrode contacts within the target structure. PMID- 17415164 TI - Dual microelectrode technique for deep brain stereotactic surgery in humans. AB - OBJECTIVE: To improve functional stereotactic microelectrode localization of small deep brain structures by developing and evaluating a recording system with two closely separated independently controlled microelectrodes. METHODS: Data were obtained from 52 patients using this dual microelectrode technique and 38 patients using the standard single microelectrode technique for subthalamic nucleus localization in patients with Parkinson's disease. RESULTS: There was a decrease in the incidence of noncontributory trajectories, defined as a single penetration made by the pair of closely spaced parallel microelectrodes, owing to microelectrode failure (from 7.2% to <1%), an improved localization and verification of nuclear borders, and a significant decrease in the number of trajectories used to localize the subthalamic nucleus from a median of three to two per initial operative side (P < 0.001). The technique also provides the novel opportunity to examine population activity by correlating the discharge between two closely spaced simultaneously recorded neurons and can be used to monitor the electrophysiological effects of local electrical stimulation or microinjections of pharmacological agents. CONCLUSION: Our experience indicates that the use of two closely spaced microelectrodes improves the utility of microelectrode localization in minimally invasive functional neurosurgery. PMID- 17415165 TI - Comparison of two techniques to postoperatively localize the electrode contacts used for subthalamic nucleus stimulation. AB - OBJECTIVE: Cerebral ventriculography (Vg) and magnetic resonance imaging (MRI) scanning are routine procedures to determine the implanted electrode placement into the subthalamic nucleus (STN) and are used in several centers that provide deep brain stimulation for Parkinson's disease patients. However, because of image distortion, MRI scan accuracy in determining electrode placement is still matter of debate. The objectives of this study were to verify the expected localization of the electrode contacts within the STN and to compare the stereotactic coordinates of these contacts determined intraoperatively by Vg with those calculated postoperatively by MRI scans. To our knowledge, this is the first study attempting to compare the "gold standard" of stereotactic accuracy (Vg) with the anatomic resolution provided by MRI scans. METHODS: Images from 18 patients with Parkinson's disease who underwent bilateral operation were used in this study. Among the 36 chronically stimulated contacts, 28 contacts (78%) were localized in the dorsolateral part of the STN. The remaining eight contacts (22%) were located more dorsally in the zona incerta, close to the upper border of the STN. RESULTS: Significant differences were found between Vg and MRI scans regarding the mediolateral x coordinate of the contacts for both left and right electrodes and regarding the right-sided anteroposterior y coordinate. No statistical difference was found for the left-sided y coordinate and the dorsoventral z coordinate for both sides. CONCLUSION: If we assume that Vg is an imaging gold standard, our results suggest that postoperative MRI scanning may induce a slight image translation compared with Vg. However, MRI scans allowed localization of most of the contacts within the STN. PMID- 17415166 TI - Graded repair of cranial base defects and cerebrospinal fluid leaks in transsphenoidal surgery. AB - OBJECTIVE: A graded approach to cerebrospinal fluid (CSF) leak repair after transsphenoidal surgery is presented. METHODS: Patients undergoing endonasal tumor removal during an 8-year period were reviewed. Intraoperative CSF leaks were classified as Grade 0, no leak observed; Grade 1, small leak without obvious diaphragmatic defect; Grade 2, moderate leak; or Grade 3, large diaphragmatic/dural defect. Cranial base repair was tailored to the leak grade as Grade 0, collagen sponge; Grade 1, two-layered collagen sponge repair with intrasellar titanium mesh buttress; Grade 2, intrasellar and sphenoid sinus fat grafts with collagen sponge overlay and titanium buttress; and Grade 3, same as Grade 2 with CSF diversion in most cases. A provocative tilt test was performed before patient discharge to assess the integrity of the CSF leak repair. Protocol modifications adopted in 2003 included an intrasellar fat graft in Grade 1 leaks with a large intrasellar dead space, frequent use of BioGlue (CryoLife, Inc., Atlanta, GA) in Grade 1, 2, and 3 leaks, and CSF diversion for all Grade 3 leaks. RESULTS: Among 668 cases in 620 patients (475 pituitary adenomas and 145 other lesions), an intraoperative CSF leak was observed in 57% of the cases: 32.5% Grade 1, 15% Grade 2, and 8.7% Grade 3. Postoperative repair failures occurred in 17 cases (2.5%), including 0.7, 3, 1, and 12% of Grade 0, 1, 2, and 3 CSF leaks, respectively. Bacterial meningitis occurred in three patients (0.45%). After protocol modifications in 2003, repair failures decreased from 4 to 1.2% (P = 0.02). CONCLUSION: A graded repair approach to CSF leaks in transsphenoidal surgery avoids tissue grafts and CSF diversion in more than 60% of patients. Protocol modifications adopted in the last 340 cases have reduced the failure rate to 1% overall and 7% for Grade 3 leaks. Provocative tilt testing before patient discharge is helpful in the timely diagnosis of postoperative CSF leaks. PMID- 17415167 TI - Wound complications associated with the use of bovine serum albumin glutaraldehyde surgical adhesive in pediatric patients. AB - OBJECTIVE: The use of dural sealants has become common in neurosurgery. Ten patients in whom a bovine albumin-glutaraldehyde combination (BioGlue; Cryolife, Inc., Kennesaw, GA) was implanted have had wound complications. The clinical experience of these patients is presented along with recommendations regarding the use of BioGlue. METHODS: All clinical information was reviewed for the patients who had wound complications after implantation of BioGlue. Data collected included diagnosis, type of original surgery, clinical presentation of the wound complication, culture results, and management. RESULTS: BioGlue was implanted in 75 patients during 77 procedures. Ten patients (13%) had wound complications. There were six boys and four girls with an average age of 5.2 years (range, 11 mo-16 yr). Three had undergone spinal procedures for detethering of the spinal cord and seven underwent craniotomies (five infratentorial, two supratentorial). Six patients presented with purulent drainage, two had periorbital cellulites, and two had swelling and fluctuance without drainage. All patients were managed operatively by washout, debridement, and removal of the BioGlue followed by a course of intravenous antibiotics. The average interval between the implantation and the debridement was 12.5 weeks (range, 2.5-28 wk). All patients had purulence around the BioGlue. Positive cultures were obtained in seven patients (Staphylococcus species in six and Streptococcus pneumoniae in one), including five who presented with wound drainage. CONCLUSION: We have found a strong association between the use of BioGlue and postoperative wound complications in our pediatric neurosurgical practice and, thus, discourage its future use. BioGlue triggers an intense acute pyogenic and chronic granulomatous inflammatory response, which is an ideal environment for bacterial growth. The bacteria may arise from the BioGlue or other foreign bodies or, more likely, from the skin if the inflammation breaches the skin surface. PMID- 17415168 TI - Cervical arthroplasty with the PRESTIGE LP cervical disc. AB - The technique for anterior cervical arthroplasty using the PRESTIGE LP (Medtronic Sofamor Danek, Memphis, TN) cervical disc is illustrated. Surgical positioning and the operative approach are demonstrated. Our methods of retractor placement and surgical exposure are discussed. The technique used for decompression and endplate preparation is presented in detail. Optimal arthroplasty device positioning is demonstrated. The surgical pearls and pitfalls of anterior cervical arthroplasty are highlighted. PMID- 17415169 TI - Resection of a benign brachial plexus nerve sheath tumor using intraoperative electrophysiological monitoring. AB - OBJECTIVE: Benign peripheral nerve sheath tumors arising from the brachial plexus are rare. Neurosurgeons often lack the clinical and surgical experience to optimize the management of these uncommon tumors. We filmed a video depicting the surgical resection of a benign peripheral nerve sheath tumor involving the brachial plexus. METHODS: An illustrative case was used to demonstrate the proper management of a brachial plexus nerve sheath tumor including the important role of intraoperative electrophysiological neuromonitoring during tumor resection. RESULTS: Using an illustrative case, we describe a systematic approach in the evaluation and surgical management of patients with a brachial plexus nerve sheath tumor. The importance of taking a thorough clinical history, performing a thorough physical examination, applying high-resolution magnetic resonance imaging techniques to visualize the pathology, and using intraoperative electrophysiological neuromonitoring during surgical exposure and resection of the tumor are stressed. Combined with appropriate postoperative treatment, these techniques minimize the risks and increase the likelihood of achieving a good clinical outcome. CONCLUSION: Brachial plexus nerve sheath tumors are challenging mass lesions that should be evaluated and surgically resected by an experienced team of physicians to optimize clinical outcome. PMID- 17415170 TI - Avoidance of carotid artery injuries in transsphenoidal surgery with the Doppler probe and micro-hook blades. AB - OBJECTIVE: Internal carotid artery (ICA) injury during sellar dural opening is a potentially catastrophic complication of transsphenoidal surgery. We describe two ICA injuries that occurred early in our endonasal transsphenoidal experience. We then describe our subsequent protocol to prevent this complication in which we use the Doppler probe for carotid localization and micro-hook blades for lateral dural opening. METHODS: All patients undergoing endonasal tumor removal were analyzed since beginning this approach in 1998. Of 631 procedures (585 patients), three patients sustained an ICA injury. RESULTS: In the first 114 procedures (105 patients) in which the Doppler probe was not used and hook blades were used infrequently, two (1.8%) ICA injuries occurred. In both cases, a right nostril approach was used and the left ICA was punctured on dural opening with a straight scalpel; both patients recovered without neurological sequelae. In the subsequent 517 procedures in which the Doppler probe and hook blades were used in all cases, one (0.19%) probable ICA injury occurred during an attempted removal of a cavernous sinus schwannoma, although there was no angiographic evidence of vascular injury. There were no ICA or other intracranial vascular injuries in the last 510 procedures for tumors not solely confined to the cavernous sinus. CONCLUSION: Cavernous carotid localization with the Doppler probe before dural opening and angled hook blades for lateral dural opening can help minimize the risk of ICA injury and are recommended for all transsphenoidal operations. Because of the wider contralateral exposure provided by the endonasal approach, the ICA contralateral to the nostril of approach is at higher risk of injury on dural opening. PMID- 17415171 TI - Neuronavigation without rigid pin fixation of the head in left frontotemporal tumor surgery with intraoperative speech mapping. AB - OBJECTIVE: Intraoperative speech mapping has evolved into the "gold standard" for neurosurgical removal of lesions near the language cortex. The integration of neuronavigation into a multimodal protocol can improve the reliability of this type of operation, but most systems require rigid fixation of the patient's head throughout the operation. This article describes and evaluates a new noninvasively attached sensor-based reference tool, which can replace rigid pin fixation of the patient's head during awake craniotomies. METHODS: The attachment technique and the resulting application accuracy were investigated under clinical conditions in 13 patients undergoing awake craniotomy with intraoperative mapping of cortical language sites. RESULTS: Spatial information was used for updating the image guidance by continuously adjusting the image planes relative to the position of the patient's head. The mean registration error achieved with this technique was 1.53 +/- 0.51 mm (fiducial registration error +/- standard deviation). The system's median application accuracy between dura opening and closure ranged from 0.83 to 1.85 mm (position error). CONCLUSION: The use of a reference sensor can replace uncomfortable pin fixation of the patient's head during navigation-supported awake craniotomies. Application accuracy is not affected by repositioning of the patient or by unavoidable head movements. Thus, this technique enables full exploitation of the benefits of navigation in a multimodal operative protocol without the need to rigidly fix the patient's head. PMID- 17415172 TI - Angled screw holes for anterior posts and a frame-positioning device for gamma knife radiosurgery: allowing for better targeting of intracranial lesions. AB - OBJECTIVE: Gamma knife radiosurgery requires frame positioning so that the treatment target is as close to the center of the frame and as low as possible to cover all of the posterior fossa contents. In this study, we report the use of two devices developed by the senior author (MWM) that facilitate these two crucial objectives in the treatment of intracranial targets using the gamma knife. METHODS: Custom front posts with threaded screw holes drilled at 5-, 10-, and 15-degree angles were created by the manufacturer at our request. A U-shaped metal device for frame positioning was designed in-house and fits into the holes at the 100-mm mark on the lateral sides of the Leksell stereotactic frame base. This allowed the positioning device to snap securely into the frame for use in positioning. The positioning device was constructed so that the lowest possible frame position would be achieved with each frame application, while avoiding collisions with the magnetic resonance imaging localizer box. RESULTS: Angled front posts allowed for pin contacts with the cranium anterior and/or superior to the superior temporal line despite a lateral or posterior position of the frame. This avoided penetration of the temporalis muscle and reduced discomfort for patients. The U-shaped metal device was used in place of the Velcro straps or ear bars routinely used for frame positioning in which the distance from the frame base to the top of the head must always be measured to avoid collisions with the localizer box. During the past 2 years, these devices have been used on a daily basis, achieving the desired results. In many cases, their use has avoided the need for frame repositioning and rescanning for targets that cannot be reached because of inexact frame positioning. CONCLUSION: A new design with angled screw holes in the front posts used for gamma knife radiosurgery allows surgeons to avoid penetration of the temporalis muscle and to maintain a perpendicular orientation of the fixation screw to the outer table of the cranium. They may also prevent mechanical creep caused by the obliquity of pin contact with the cranium and resulting loss of torque. We also present a simple device that may be useful in frame positioning. The device ensures a frame position as low as possible without the need for measurement at the time of frame positioning. PMID- 17415173 TI - Neuronavigation for arteriovenous malformation surgery by intraoperative three dimensional ultrasound angiography. AB - OBJECTIVE: Neuronavigational devices have traditionally used preoperative imaging with limited possibilities for adjustment to brain shift and intraoperative manipulation of the surgical lesions. We have used an intraoperative imaging and navigation system that uses navigation on intraoperatively acquired three dimensional ultrasound data, as well as preoperatively acquired magnetic resonance imaging scans and magnetic resonance angiograms. The usefulness of this system for arteriovenous malformation (AVM) surgery was evaluated prospectively. METHODS: Nine consecutive patients with Spetzler Grade 1 (n = 3), 2 (n = 3), 3(n = 2) or 4 (n = 1) AVMs underwent operation using this intraoperative imaging and navigation system. The system provides real-time rendering of three-dimensional angiographic data and can visualize such projections in a stereoscopic (virtual reality) manner using special glasses. The experiences with this technology were analyzed and the outcomes assessed. Angiographic reconstructions of three dimensional images were obtained before and after resection. RESULTS: Conventional navigation on the basis of preoperative magnetic resonance angiography was helpful to secure positioning of the bone flap; stereoscopic visualization of the same data represented a powerful means to construct a mental three-dimensional picture of the extent of the AVM and the feeder anatomy even before skin incision. Intraoperative ultrasound corresponded well to the intraoperative findings and allowed confirmation of feeding vessels in surrounding gyri and rapid identification of the perinidal dissection planes, regardless of brain shift. The latter feature was particularly helpful because the intraoperative navigational identification of surgical planes leads to minimal exploration into the nidus or dissection at a greater distance from the malformation. Application of the system was thought to increase surgical confidence. In two patients, postresection ultrasound prompted additional nidus removal. Ultrasound angiography seemed to allow some degree of resection control, although its sensitivity was not thought to be sufficient. All AVMs were radically removed without new permanent morbidity. CONCLUSION: The complexities of handling the pathological vessels of AVMs were ameliorated by intraoperative three-dimensional ultrasound and navigation because the three-dimensional outline of the vasculature (feeders, nidus, and draining veins) provided a means to adapt resection strategies, define dissection planes, and interpret intraoperative findings. It is difficult to provide a scientifically valid definition of "added value." However, in our experience, the added confidence and the improved mental image of the lesion that resulted from this technology improved the quality and flow of surgery. PMID- 17415174 TI - Combined x-ray and magnetic resonance imaging facility: application to image guided stereotactic and functional neurosurgery. AB - OBJECTIVE: To assess the feasibility of a hybrid imaging setup combining x-ray and magnetic resonance imaging (MRI) in the setting of both stereotactic and functional neurosurgery. METHODS: A combined x-ray and MRI scanning facility with a trolley system for a fast patient transfer between both modalities was installed in a neurosurgical setting. A registration algorithm for fusion of MRI scans and x-ray images was derived for augmentation of fluoroscopic x-ray projection images with MRI scan data, such as anatomic structures and planned probe trajectories. Phantom measurements were obtained between both modalities for estimation of registration accuracy. Practical application of our system in stereotactic and functional neurosurgery was tested in brachytherapy, deep brain stimulation, and motor cortex stimulation. RESULTS: Phantom measurements yielded a mean spatial deviation of 0.7 +/- 0.3 mm with a maximum deviation of 1.1 mm for MRI scans versus x-rays. Augmentation of x-ray images with MRI scan data allowed intraoperative verification of the planned trajectory and target in three types of neurosurgical procedures: positioning iodine seeds in brachytherapy in one case with cerebellar metastasis, placement of electrodes for deep brain stimulation in two cases of advanced Parkinson's disease, and placement of an epidural grid for motor cortex stimulation in two cases of intractable pain. CONCLUSION: Combined x-ray and MRI-guided stereotactic and functional neurosurgery is feasible. Augmentation of x-ray projection images with MRI scan data, such as planned probe trajectories and MRI scan segmented anatomic structures may be beneficial for probe guidance in stereotactic and functional neurosurgery. PMID- 17415175 TI - Intraoperative application of thermography in extracranial-intracranial bypass surgery. AB - OBJECTIVE: The extracranial-intracranial bypass may have the potential to improve hemodynamic cerebral ischemia caused by occlusive diseases of the main cerebral arteries. Intraoperative confirmation of effective distribution of blood flow via the donor arteries to the involved region will assure a successful bypass surgery. METHODS: Infrared thermography was used to measure the temperature of the cortical surface at the operative field. Regional cerebral blood flow (rCBF) was measured with a laser Doppler flow meter. Changes in the cortical surface temperature before and after temporary occlusion of the bypass were compared with changes in rCBF values in the corresponding sites. RESULTS: Thermographic examination demonstrated a heterogeneous increase of cortical surface temperature caused by the blood flow via the extracranial-intracranial bypass and was closely related to rCBF changes. CONCLUSION: Thermography is useful not only to demonstrate the distribution of blood flow through the extracranial-intracranial bypass but also to quantitatively evaluate the rCBF changes in the operative field. PMID- 17415176 TI - A novel, inexpensive method of image coregistration for applications in image guided surgery using augmented reality. AB - OBJECTIVE: Augmented reality (AR) is a technique in which an overlay of a virtual image to a live picture is performed to create a new image in which both original images coexist as a single image. This results in the visualization of internal structures through overlying tissues. The objective was to describe an easy, inexpensive, and successful method to coregister with AR in an image-guided surgery setting using the resources at hand. METHODS: Cortical information was obtained with a volumetric acquisition of 200 0.8-mm thick, cerebral magnetic resonance imaging scans in an axial T1-weighted sequence. For the venous anatomy, a contrast phase at 7 mm/s velocity was used. This data was reconstructed in a three-dimensional fashion using MRIcro software (v. 1.37, freeware, courtesy of Chris Rorden) and was overlaid to a digital image of the cerebral cortex either pre- or intraoperatively. RESULTS: Eight patients were studied. There was an adequate coregistration in seven of the patients as confirmed by intraoperative ultrasound, frame-based stereotaxy, or obvious anatomic homology between the three-dimensional magnetic resonance imaging scan virtual reconstruction and the live image obtained during surgery. AR was not possible in one case of a cerebellar lesion. CONCLUSION: AR coregistration capabilities are adequate when revised by other intraoperative guidance devices. When performed with "freeware" software and conventional digital cameras, it is relatively inexpensive, which makes it a potential tool for surgical planning and noncontinuous intraoperative guidance in neurosurgery. Its largest drawbacks are the inability to function in deep-seated lesions and its lack of tracking devices, which gives it a noncontinuous coregistration nature. PMID- 17415177 TI - Optically neuronavigated ultrasonography in an intraoperative magnetic resonance imaging environment. AB - OBJECTIVE: To develop a clinically useful method that shows the corresponding planes of intraoperative two-dimensional ultrasonography and intraoperative magnetic resonance imaging (MRI) scans determined with an optical neuronavigator from an intraoperative three-dimensional MRI scan data set, and to determine the qualitative and the quantitative spatial correspondence between the ultrasonography and MRI scans. METHODS: An ultrasound probe was interlinked with an ergonomic and MRI scan-compatible ultrasonography probe tracker to the optical neuronavigator used in a low-field intraoperative MRI scan environment for brain surgery. Spatial correspondence measurements were performed using a custom-made ultrasonography/MRI scan phantom. In this work, instruments to combine intraoperatively collected ultrasonography and MRI scan data with an optical localization method in a magnetic environment were developed. The ultrasonography transducer tracker played an important role. Furthermore, a phantom for ultrasonography and MRI scanning was produced. This is the first report, to our knowledge, regarding the possibility of combining the two most important intraoperative imaging modalities used in neurosurgery, ultrasonography and MRI scanning, to guide brain tumor surgery. RESULTS: The method was feasible and, as shown in an illustrative surgical case, has direct clinical impact on image guided brain surgery. The spatial deviation between the ultrasonography and the MRI scans was, on average, 1.90 +/- 1.30 mm at depths of 0 to 120 mm from the ultrasonography probe. CONCLUSION: The overall result of this work is a unique method to guide the neurosurgical operation with neuronavigated ultrasonography imaging in an intraoperative MRI scanning environment. The relevance of the method is emphasized in minimally invasive neurosurgery. PMID- 17415178 TI - Intraoperative ultrasound using phase inversion harmonic imaging: first experiences. AB - OBJECTIVE: To study the feasibility of intraoperative ultrasound using the phase inversion harmonic imaging (PIHI) technique. METHODS: Eight patients with intracranial middle cerebral artery aneurysms and five patients with arteriovenous malformations were studied after written informed consent. A first ultrasound study was performed through the intact dura mater after cranial trepanation to assess the pathology, its feeding artery, and downstream segments. A second ultrasound study was performed immediately after intervention to monitor the success of the procedure. All patients were studied using a Siemens Sonoline Antares ultrasound machine (Siemens Medical Solutions USA, Inc., Malvern, PA) before and after intravenous administration of an ultrasound contrast agent (Optison; GE Healthcare, Milwaukee, WI). Other than conventional brightness mode, PIHI is sensitive to the nonlinear acoustic response of tissue, and especially to ultrasound contrast agent microbubbles. The latter enables contrast-specific vascular imaging. RESULTS: PIHI provided anatomically detailed information. In combination with an ultrasound contrast agent, angiography-like views of the vascular pathologies, including their surrounding vessels, could be obtained. Flow velocities in afferent and downstream vascular segments, as well as inside the pathology, could be assessed. Flow dynamics inside the aneurysm sac or the arteriovenous malformation could be studied in real-time. Postintervention, contrast-enhanced PIHI could be used to immediately monitor the success of the surgical procedure. CONCLUSION: PIHI enables intraoperative visualization and morphological assessment of neurovascular pathologies, such as middle cerebral artery aneurysms or arteriovenous malformations. In combination with an ultrasound contrast agent, the flow dynamics of these lesions can be displayed in real-time. PMID- 17415179 TI - Traumatic loading of the Bryan cervical disc prosthesis: an in vitro study. AB - OBJECTIVE: The Bryan disc prosthesis (Medtronic Sofamor Danek, Memphis, TN) relies on a precise fit between the device and the vertebral endplates to provide immediate stability after cervical arthroplasty. The safety of the cervical arthroplasty in the setting of trauma is unknown. We compare the segmental strength of the normal cervical spine and the cervical spine after single-level arthroplasty. METHODS: Fifteen cadaveric cervical spines with arthroplasty placed at the C5-C6 level were compared with 16 intact cadaveric controls. A pure moment was applied to induce flexion, extension, or axial rotation until the segment failed. RESULTS: The prosthesis provided 63, 45, and 69% of the strength of a normal spine during flexion, extension, and rotation, respectively. There were no cases of prosthesis expulsion. CONCLUSION: After insertion of the Bryan disc, the remaining ligamentous tissues provide adequate acute stability to the spine. PMID- 17415180 TI - Neuronavigational endoscopic endonasal sellar and parasellar surgery using a 2-mm diameter lens rigid-rod endoscope: a cadaver study. AB - OBJECTIVE: Most of the endoscopes used for endonasal transsphenoidal surgery use 4-mm diameter lenses. The applicability of a newly developed neuroendoscope with a lens diameter of only 2 mm was tested in endonasal transsphenoidal pituitary surgery. METHODS: The newly developed rigid-rod neuroendoscope with a 2-mm lens and an endoscope with a 4-mm lens were coupled with a navigation system and used for this comparative study. Comparison between the views obtained with these two devices was performed in a model and in formalin-fixed cadaver heads. A pure endonasal approach was used to reach and explore the sellar and parasellar regions. The navigation system was used to locate the same position in both lenses for image comparison. RESULTS: The sellar and parasellar regions could be reached and explored using the new endoscope with the 2-mm lens and an oval shaped irrigation and suction channel. The visual field appeared to be reduced compared with that of the 4-mm lens. However, this reduction was compensated by greater mobility and easier introduction and maneuvering of the instruments at the sellar level. Reduced image size and brightness were also found using the 2 mm lens compared with the 4-mm lens. These differences could be overcome by increasing the amount of light and enlarging the image but with subsequent reduction in image resolution. CONCLUSION: The small diameter of this neuroendoscope resulted in good maneuverability and maintained a fine quality of vision. Children and patients with small nostrils are good candidates for the use of such a device. PMID- 17415182 TI - Symptomatic mass effect of a hypertrophied pericranial flap after repair of a dural defect: case report. AB - OBJECTIVE: To detail a potential complication of the use of vascularized pericranial flaps in the repair of dural defects, namely, flap hypertrophy secondary to venous engorgement. CLINICAL PRESENTATION: A 23-year-old man with a left parietal lobe hemangiopericytoma underwent a craniotomy for tumor resection. The resultant dural defect was repaired with a vascularized pericranial flap. On postoperative Day 3, the patient developed headache, confusion, aphasia, and right upper extremity apraxia. Imaging revealed an extra-axial collection at the craniotomy site; on reexploration, a swollen, engorged pericranial flap causing mass effect was found. INTERVENTION: The pericranial flap was excised. CONCLUSION: In this case, hypertrophy of the vascularized pericranial flap is hypothesized to have occurred because of venous congestion, possibly secondary to restriction of venous outflow by the overlying bone flap. PMID- 17415183 TI - Chronic subdural hematoma associated with subdural rhabdomyosarcoma: case report. AB - OBJECTIVE: Subdural rhabdomyosarcoma is very rare, and even more unusual is the association between these sarcomas and chronic subdural hematoma. In this report, we present a case of subdural rhabdomyosarcoma that developed in a chronic refractory subdural hematoma. CLINICAL PRESENTATION: A 40-day-old boy was admitted with coma and intractable seizure after a head trauma caused by a fall. A subdural hematoma was detected, which later became chronic and refractory to treatment. INTERVENTION: The patient repeatedly underwent different surgical interventions over 2 years. Finally, extensive bilateral frontotemporoparietal craniotomy was performed at the age of 30 months, which revealed a huge subdural mass. The mass and membranes were resected. Pathological examination confirmed the diagnosis of embryonal-type rhabdomyosarcoma. Radiation therapy was performed, but the tumor recurred and the child died at the age of 3 years. CONCLUSION: Whether the subdural hematoma or the tumor was the initial pathology is debatable. We think the sarcoma probably developed gradually because of chronic inflammation and proliferation in the subdural collection. PMID- 17415184 TI - Superior sagittal sinus thrombosis caused by calvarial tuberculosis: case report. AB - OBJECTIVE: Tuberculosis affects the central nervous system in various ways but has not been reported to cause venous sinus thrombosis. In this case report, extensive calvarial tuberculosis caused compression and occlusion of the superior sagittal sinus in an adult man. Early diagnosis combined with surgical and medical treatment resulted in cure of the disease. CLINICAL PRESENTATION: A 34 year-old man presented with a nonhealing sinus in the frontal area after a trivial injury without any neurological deficits or features of raised intracranial pressure. Magnetic resonance imaging scans showed evidence of extensive bilateral extradural granulations, bone destruction, and thrombosis of the anterior half of superior sagittal sinus. INTERVENTION: The affected bone and extradural granulations were surgically excised. Histopathology showed tuberculous osteomyelitis, and the patient received anti-tuberculous treatment, after which he had a good recovery. CONCLUSION: Calvarial tuberculosis as a cause of sagittal sinus thrombosis is a rare condition and is reported here for the first time, to our knowledge. Surgical excision of the compressing granulations followed by medical treatment is curative without sequelae if the intervention is performed before the disease extends intradurally. PMID- 17415185 TI - Neuromuscular choristoma of the oculomotor nerve: case report. AB - OBJECTIVE: Neuromuscular choristomas (NMC) are rare benign tumors of the peripheral nerves. We report an NMC affecting the oculomotor nerve. CLINICAL PRESENTATION: An 18-year-old girl presented with long-standing intermittent retro orbital pain and oculomotor paresis. Magnetic resonance imaging scans demonstrated a small nodular lesion on the left oculomotor nerve, similar to the findings for a schwannoma. INTERVENTION: The tumor was resected with the parental oculomotor nerve, which was reconstructed using a peroneal nerve graft. Postoperatively, the patient became pain-free, and her oculomotor function partially recovered. Histologically, the lesion consisted of well-differentiated smooth muscle fibers intermingled with mature nerve elements consistent with the diagnosis of an NMC, although the possibility of leiomyoma in this rare location was not excluded completely. CONCLUSION: NMC may need histological confirmation for diagnosis if they occur in the intracranial space. The resection is feasible, and the function of the affected nerve can be at least partially restored with the nerve reconstruction. PMID- 17415186 TI - Lymphangioma of the clivus: case report. AB - OBJECTIVE: Different clival lesions have been reported in the literature previously. The lymphangioma reported in the present case seems to be the first lesion localized in the clivus. CLINICAL PRESENTATION: The patient presented with a history of diplopia and headache. Imaging studies showed a lytic lesion at the cranial base. INTERVENTION: Transsphenoidal excision of the lesion resulted in total resolution of the patient's complaints. CONCLUSION: Lymphangioma must be kept in mind for the differential diagnosis of lytic lesions of the cranial base. Surgical removal leads to immediate improvement of the symptoms. PMID- 17415187 TI - Extracranial nasopharyngeal craniopharyngioma: case report. AB - OBJECTIVE: Craniopharyngiomas (CPs) are benign tumors that almost always occur in a suprasellar location, making complete resection difficult and often necessitating radiotherapy. A case of CP presenting in an unusual location in an 8-year-old boy highlights the goals of CP treatment. CLINICAL PRESENTATION: An 8 year-old boy sought treatment for symptoms of nasal obstruction and snoring. He also had a history of mild developmental delay, and his father had a thyroglossal duct cyst resected in his own youth. After tonsillectomy and adenoidectomy failed to improve the patient's symptoms, nasal endoscopy and biopsy revealed an intranasal CP. After this treatment, he experienced chronic thin brown nasal discharge. Magnetic resonance imaging further revealed tumor invading the sphenoid body and the clivus and that the tumor had no intracranial extension. INTERVENTION: The child underwent surgical resection via a Le Fort I osteotomy approach. Complete resection was accomplished based on intraoperative frozen section pathological examination and postoperative magnetic resonance imaging results. CONCLUSION: CP with no intracranial extension is a very rare but benign tumor. We recommend vigorous attempts to resect such tumors completely to minimize the chance of recurrence and the possible need for radiotherapy. Although radiotherapy controls CP growth quite well, it has its own risks that should be avoided if possible. Cranial base techniques may facilitate total resection. PMID- 17415188 TI - Symptomatic Chiari type I malformation after radiation therapy in an infant: case report. AB - OBJECTIVE: One previous case report has described the development of a Chiari I malformation in a child after fractionated radiation therapy to the anterior cranial base. The authors present an additional case of a 17-month-old girl treated for an isolated, malignant, rhabdoid tumor of the right neck with neoadjuvant chemotherapy, surgical exploration, and radiation therapy. CLINICAL PRESENTATION: The patient developed a Chiari I malformation and cervical syringomyelia 1.5 years after the treatment. The patient experienced two episodes of unresponsiveness before diagnosis, and the malformation was then identified on magnetic resonance imaging scans. INTERVENTION: Suboccipital decompressive craniectomy, and C1 laminectomy with autologous duraplasty were performed. RESULTS: Postoperative magnetic resonance imaging scans revealed reconstitution of cerebrospinal fluid at the foramen magnum and ascent of the cerebellar tonsils. The patient remains asymptomatic 5.5 years after treatment. CONCLUSION: The importance of this association is discussed in the context of the increasing use of radiation therapy in young children and infants. PMID- 17415192 TI - The development of spinal neurosurgery: a historical perspective. PMID- 17415193 TI - The evolving presence of spinal neurosurgery in the spectrum of neurological surgery. PMID- 17415194 TI - The evolving presence of spinal neurosurgery in the spectrum of neurological surgery. PMID- 17415195 TI - Magnetic resonance imaging-guided focused ultrasound for thermal ablation in the brain: a feasibility study in a swine model. AB - INTRODUCTION: Magnetic resonance imaging (MRI)-guided focused ultrasound is a novel technique that was developed to enable precise, image-guided targeting and destruction of tumors by thermocoagulation. The system, ExAblate2000, is a focused ultrasound delivery system embedded within the MRI bed of a conventional diagnostic MRI scanner. The device delivers small volumetric sonications from an ultrasound phased array transmitter that converge energy to selectively destroy the target. Temperature maps generated by the MRI scanner verify the location and thermal rise as feedback, as well as thermal destruction. To assess the safety, feasibility, and precision of this technology in the brain, we have used the ExAblate system to create predefined thermal lesions in the brains of pigs. METHODS: Ten pigs underwent bilateral craniectomy to provide a bone window for the ultrasound beams. Seven to 10 days later, the animals were anesthetized and positioned in the ExAblate system. A predefined, 1-cm frontal para ventricular region was delineated as the target and treated with multiple sonications. MRI was performed immediately and 1 week after treatment. The animals were then sacrificed and the brains removed for pathological study. The size of individual sonication points and the location of the lesion were compared between the planned dose maps, posttreatment MRI scans, and pathological specimen. RESULTS: High-energy sonications led to precise coagulation necrosis of the specified targets as shown by subsequent MRI, macroscopic, and histological analysis. The thermal lesions were sharply demarcated from the surrounding brain with no anatomic or histological abnormalities outside the target. CONCLUSION: MRI-guided focused ultrasound proved a precise and an effective means to destroy anatomically predefined brain targets by thermocoagulation with minimal associated edema or damage to adjacent structures. Contrast-enhanced T1-, T2-, and diffusion-weighted MRI scans may be used for real-time assessment of tissue destruction. PMID- 17415196 TI - The potential of ferumoxytol nanoparticle magnetic resonance imaging, perfusion, and angiography in central nervous system malignancy: a pilot study. AB - OBJECTIVE: Ferumoxytol, an iron oxide nanoparticle that targets phagocytic cells, can be used in magnetic resonance imaging of malignant brain tumors and can be administered as a bolus, allowing dynamic imaging. Our objectives were to determine the optimum time of delayed contrast enhancement of ferumoxytol, and to compare ferumoxytol and gadolinium contrast agents for magnetic resonance angiography and perfusion. METHODS: Twelve patients with malignant brain tumors underwent serial magnetic resonance imaging multiple times up to 72 hours after ferumoxytol injection at both 1.5 and 3-T. The enhancement time course was determined for ferumoxytol and compared with a baseline gadolinium scan. Perfusion, time-of-flight and dynamic magnetic resonance angiography and T1 weighted scans were compared for the two agents. RESULTS: The lesions were detectable at all field strengths, even with an intraoperative 0.15-T magnet. Maximal ferumoxytol enhancement intensity was at 24 to 28 hours after administration, and the enhancing volume subsequently expanded with time into a non-gadolinium-enhancing, high T2-weighted signal region of tumor-infiltrated brain. Dynamic studies were assessed with both agents, indicating early vascular leak with gadolinium but not with ferumoxytol. CONCLUSION: Our most important finding was that gadolinium leaks out of blood vessels early after injection, whereas ferumoxytol stays intravascular in the "early" phase, thereby increasing the accuracy of tumor perfusion assessment. As a magnetic resonance imaging contrast agent, ferumoxytol visualizes brain tumors at all field strengths evaluated, with delayed enhancement peaking at 24 to 28 hours after administration. PMID- 17415197 TI - Treatment options for third ventricular colloid cysts: comparison of open microsurgical versus endoscopic resection. AB - OBJECTIVE: We retrospectively reviewed our experience treating third ventricular colloid cysts to compare the efficacy of endoscopic and transcallosal approaches. METHODS: Between September 1994 and March 2004, 55 patients underwent third ventricular colloid cyst resection. The transcallosal approach was used in 27 patients; the endoscopic approach was used in 28 patients. Age, sex, cyst diameter, and presence of hydrocephalus were similar between the two groups. RESULTS: The operating time and hospital stay were significantly longer in the transcallosal craniotomy group compared with the endoscopic group. Both approaches led to reoperations in three patients. The endoscopic group had two subsequent craniotomies for residual cysts and one repeat endoscopic procedure because of equipment malfunction. The transcallosal craniotomy group had two reoperations for fractured drainage catheters and one operation for epidural hematoma evacuation. The transcallosal craniotomy group had a higher rate of patients requiring a ventriculoperitoneal shunt (five versus two) and a higher infection rate (five versus none). Intermediate follow-up demonstrated more small residual cysts in the endoscopic group than in the transcallosal craniotomy group (seven versus one). Overall neurological outcomes, however, were similar in the two groups. CONCLUSION: Compared with transcallosal craniotomy, neuroendoscopy is a safe and effective approach for removal of colloid cysts in the third ventricle. The endoscope can be considered a first-line treatment for these lesions, with the understanding that a small number of these patients may need an open craniotomy to remove residual cysts. PMID- 17415198 TI - Response of C-reactive protein after craniotomy for microsurgery of intracranial tumors. AB - OBJECTIVE: C-reactive protein (CRP) is used as an indicator of inflammatory processes. However, its expression is unspecific and will increase after surgery. Without normative data on its regular course after craniotomy, postoperative infection can barely be detected or excluded. METHODS: CRP was measured for 10 days in 46 patients who underwent elective craniotomy for microsurgery of intracranial tumors. RESULTS: After craniotomy, CRP rapidly increased to reach a peak mean value of 32.43 +/- 38.02 mg/l (P < 0.001) on the second postoperative day. In 39 patients (85%), CRP reached its maximum level within the first 2 days. From Days 3 to 5 after surgery, mean CRP values constantly and significantly declined (P < 0.001) to arrive at a mean of 6.67 +/- 10.80 mg/l) on the fifth postoperative day. On Day 4, the mean CRP level returned to below one-third of the peak value (10.63 +/- 17.08 mg/l). On an individual basis, this was true for 34 patients (74%). During the period of initial increase of CRP until the second postoperative day, there was no significant correlation between CRP and erythrocyte sedimentation rate, body temperature, hemoglobin, hematocrit, red cell count, platelet count, and white cell count. CRP increase was more pronounced with intrinsic brain tumors and was not correlated to the degree of malignancy. CONCLUSION: Because CRP can be considerably increased in patients for 4 days after regular intracranial surgery, its diagnostic value during this period is limited. Only prolonged elevation or a secondary increase may indicate an ongoing infection. PMID- 17415199 TI - Endovascular coil embolization of ruptured and unruptured posterior circulation aneurysms: review of a 10-year experience. AB - OBJECTIVE: Treatment of posterior circulation aneurysms poses a great technical challenge for the practicing neurosurgeon. The advent of endovascular techniques has made such treatment more feasible. We report our experience with the endovascular management of ruptured and unruptured posterior circulation aneurysms during the past 10 years. METHODS: A retrospective analysis was performed on all patients with posterior circulation aneurysms undergoing endovascular treatment at Jefferson Hospital for Neuroscience between July 1995 and December 2005. This yielded 275 patients (67 men and 208 women). The degree of aneurysm occlusion was determined by the operating endovascular neurosurgeon at the time of the procedure. Successful embolization was defined as greater than 95% occlusion of the dome without any coil prolapsing into the parent vessel. Clinical outcome was evaluated using the modified Glasgow Outcome Scale. Clinical follow-up data was obtained for 262 patients (95.3%); the follow-up period ranged from 1 to 94 months (mean, 31.8 mo for procedures performed before 2004 and 13.3 mo for procedures performed during 2004 and 2005). Angiographic follow-up data was obtained for 224 patients (84.8%) for periods ranging from 6 to 94 months (mean, 31.3 mo for procedures performed before 2004 and 13.7 mo for procedures performed during 2004 and 2005). RESULTS: Based on the Hunt and Hess grading scale, the patient population included 106 patients (38.5%) with unruptured aneurysms, 43 patients (15.6%) with Grade I aneurysms, 16 patients (5.8%) with Grade II aneurysms, 56 patients (20.5%) with Grade III aneurysms, and 54 patients (19.6%) with Grade IV aneurysms. The locations of the posterior circulation aneurysms included 189 (68.7%) in the basilar apex or posterior cerebral artery, 23 (8.4%) in the basilar trunk/anterior inferior cerebellar artery, 22 (8%) in the superior cerebellar artery, and 41 (14.9%) in the vertebral artery or posterior inferior cerebellar artery. Of the 275 patients, 208 (76%) were women and 67 (24%) were men. The mean age at the time of treatment was 53.9 years (range, 7-90 yr). Of all patients treated, 237 patients (87.8%) had successful embolization (>95% occlusion of the dome). On angiographic follow-up, 55 patients (24.5%) developed recanalization of at least 5%. Retreatment was required in 11 patients (4.9%; 0.01%/patient yr) and rehemorrhage occurred in three patients (1.1%; 0.003%/patient yr). Clinical follow-up was graded using the modified Glasgow Outcome Scale (mGOS) and revealed 229 patients (87.4%) in the mGOS I category, 12 patients (4.6%) in the mGOS II category, eight patients (3%) in the mGOS III category, two patients (0.8%) in the mGOS IV category, and 11 patients (4.2%) were deceased (mGOS V). Clinically significant vasospasm requiring angioplasty occurred in 11 patients (6.5%) with subarachnoid hemorrhage, and 120 patients (71%) with subarachnoid hemorrhage required ventricular shunts. Complications causing clinical morbidity occurred in 14 patients (5.1%) and ranged from postoperative ischemia to recurrent subarachnoid hemorrhage. Of all clinical factors evaluated, Hunt and Hess grade was the strongest predictor of good clinical outcome (P < 0.0001). CONCLUSION: Endovascular coil embolization of posterior circulation aneurysms is an effective treatment in the short term but is associated with recurrence, which requires close surveillance, possible retreatment, and can, albeit very rarely, lead to rehemorrhage. Future technological advancements such as the development of biologically active coils will be essential in the permanent obliteration of aneurysms. PMID- 17415200 TI - The effects of diffuseness and deep perforating artery supply on outcomes after microsurgical resection of brain arteriovenous malformations. AB - OBJECTIVE: Diffuse arteriovenous malformations (AVM) have non-compact niduses, irregular margins, and intervening brain parenchyma. Deep perforating arteries often contribute to the ragged border of these diffuse AVMs. We hypothesized that diffuseness and deep perforator supply increase the difficulties and risks associated with microsurgical AVM resection. METHODS: Diffuseness was quantified using computer-generated outlines of AVMs on angiograms, contour plots with varying image intensities, and calculations of nidus area-intensity profiles. Diffuse AVMs had nonlinear area-intensity profiles with high transition intensities ([I*] greater than 0.5). A consecutive series of 304 patients who were treated with microsurgical AVM resection over a period of 7.8 years was analyzed, along with quantification of diffuseness in a subset of 103 consecutive patients. Neurological outcomes were assessed by using the Modified Rankin Scale, and logistic regression analysis was used to identify predictors of deterioration and poor outcome at late follow-up evaluation. RESULTS: Diffuse niduses were observed in 25% of patients, and 18% of patients had deep perforating artery supply. Patients with compact AVMs were more likely to have good outcomes or overall improvement (88 and 87%, respectively) than patients with diffuse AVMs (65 and 54%, respectively) (P = 0.008 and P < 0.001, respectively). Similarly, absence of deep perforator supply was associated with good outcomes or improvement in 85 and 78% of patients, respectively, compared with 63 and 64% of patients, respectively, in patients with deep perforator supply (P < 0.001 and P = 0.028, respectively). By logistic regression analysis, diffuseness and deep perforator supply were both associated with significant increases in surgical risk. CONCLUSION: Diffuseness and deep perforating artery supply are subtle features of an AVM that predict worse outcomes after microsurgical resection. Diffuseness makes surgical planes more difficult to determine and follow, whereas deep perforators are friable, poorly visualized, and located in eloquent white matter tracts. The Spetzler-Martin grading scale does not directly account for these two features; however, they should be considered carefully when making treatment recommendations to patients with AVMs. PMID- 17415201 TI - Cognitive outcome after aneurysmal subarachnoid hemorrhage: time course of recovery and relationship to clinical, radiological, and management parameters. AB - OBJECTIVE: Although many patients show a satisfactory physical outcome after aneurysmal subarachnoid hemorrhage (SAH), disabling cognitive dysfunction may still be present. This study focuses on the time course of cognitive recovery during the first year after aneurysmal SAH, and relates the neuropsychological test results to clinical, radiological, and management parameters. METHODS: Thirty-two patients were followed prospectively with neuropsychological examinations at 3, 6, and 12 months after SAH. Test results were compared with clinical entry variables, management variables, and pre- and postoperative radiological findings. RESULTS: The time course of cognitive recovery after aneurysmal SAH is heterogeneous, with motor and psychomotor functions recovering within the first 6 months, whereas verbal memory did not improve significantly until at least 6 months after the ictus. Clinical and radiological parameters reflecting the impact of the bleed were related to memory function, intelligence, and aphasia. The site of aneurysm and mode of treatment could not be linked to neuropsychological outcome. The time length of volume-controlled mechanical ventilation as a reflector of the aggregated consequences of being subjected to an aneurysm rupture correlated with both motor and psychomotor functioning and memory performance, predominantly 6 to 12 months after SAH, but was not linked to intelligence or aphasia. CONCLUSION: The various cognitive functions have different time courses of recovery, with verbal memory requiring the longest time. Parameters reflecting the impact of the bleed and patient management can be linked to neuropsychological outcome. PMID- 17415202 TI - Predictors of cerebral infarction in patients with aneurysmal subarachnoid hemorrhage. AB - OBJECTIVE: Cerebral infarction would be expected to be associated with poor outcome after aneurysmal subarachnoid hemorrhage (SAH), although there are few data on which to base this assumption. The goals of this study were to determine the impact of cerebral infarction on outcome and to examine predictors of infarction in these patients. METHODS: Univariate and multivariable statistical methods were used to examine the impact of cerebral infarction on the Glasgow Outcome Scale score 3 months after SAH among 3567 patients entered into four prospective, randomized, double-blind, placebo-controlled trials of tirilazad conducted in neurosurgical centers around the world between 1991 and 1997. Patient demographics, clinical variables, radiographic characteristics, and treatment variables associated with cerebral infarction were also determined by the same methods. RESULTS: Seven hundred and seven (26%) out of 2741 patients with complete data had cerebral infarction on computed tomographic scans 6 weeks after SAH. Multivariable logistic regression showed that cerebral infarction increased the odds of unfavorable outcome by a factor of 5.4 (adjusted odds ratio, 5.4; 95% confidence interval, 4.2-6.8; P < 0.0001), which was a higher odds ratio than all other factors associated with outcome. The proportion of explained variance in outcome was also highest for cerebral infarction and accounted for 39% of the explained variance. Multivariable analysis found that cerebral infarction was significantly associated with increasing patient age, worse neurological grade on admission, history of hypertension or diabetes mellitus, larger aneurysm, use of prophylactically or therapeutically induced hypertension, temperature more than 38 degrees C 8 days after SAH, and symptomatic vasospasm. CONCLUSION: Cerebral infarction was strongly associated with poor outcome after aneurysmal SAH. The most important potentially treatable factor associated with infarction was symptomatic vasospasm. PMID- 17415203 TI - Tumor necrosis factor-alpha gene and cerebral aneurysms. AB - OBJECTIVE: The pathogenesis of intracranial aneurysms is still uncertain. In addition to atherosclerosis, immunological factors may play a role in the disease. Recent studies have suggested that tumor necrosis factor-alpha (TNF alpha), one of the main proinflammatory cytokines, may play a key role in the formation and rupture of cerebral aneurysms. The purpose of this study is to evaluate the association of a functionally active polymorphism (-308 G or =7 mm) and poor lateral wall motion (1 or 2 of 5, with 3 being rated as average) measured by multiview videofluoroscopy and nasendoscopy. RESULTS: The mean preoperative score on the speech and voice rating scale was 10.5 (range, 4 to 13), with a mean postoperative score of 1.9 (range, 0 to 8) following Furlow palatoplasty and sphincter pharyngoplasty in 13 patients. Two patients required an additional surgical procedure to achieve complete correction of velopharyngeal incompetence without nasal airway obstruction, to achieve a final mean score of 0.8 (range, 0 to 2) among the 12 patients who completed surgical management, achieving a highly significant reduction in nasality (p < 0.0001). CONCLUSIONS: Patients with velopharyngeal incompetence who have a black hole on nasendoscopy consisting of a large velopharyngeal gap and poor lateral wall motion are at high risk for recurrent velopharyngeal incompetence or nasal airway obstruction following surgical management. Initial treatment with a Furlow palatoplasty and sphincter pharyngoplasty has a high rate of success in these patients and does not preclude further surgical correction if needed, with minimal risk of nasal airway compromise. PMID- 17415249 TI - Identifying reproducible patterns of calvarial dysmorphology in nonsyndromic sagittal craniosynostosis may affect operative intervention and outcomes assessment. AB - BACKGROUND: The authors tested the premise that there are four distinctive patterns of calvarial dysmorphology in nonsyndromic sagittal craniosynostosis that can be reproducibly recognized. METHODS: Twenty-nine computed tomographic scan data sets of infants met the following criteria: nonsyndromic sagittal craniosynostosis, age younger than 12 months, and satisfactory computed tomographic data. Osseous reformations were constructed in the anteroposterior, right lateral, and vertex projections for each patient. From these images, four templates--coronal constriction, occipital protuberance, bifrontal bossing, and bitemporal protrusion--were selected as prototypes of the specific dysmorphologies the authors observed in patients with sagittal craniosynostosis. Four residents assigned the 29 calvarial image sets to one of the four templates or, if they were unable to do so, to the group "other." The sortings were then assessed for clustering. The same patient computed tomographic data were reformatted with osseous color images, which were then sorted according to template group by eight senior craniofacial surgeons, who repeated the task approximately 3 months later. The repeatability and assessment of clustering of image sets using the templates was evaluated. RESULTS: In the residents' pilot study, 41 percent (12 of 29) of patients had 100 percent concordance rates, 31 percent (nine of 29) had 75 percent concordance, 24 percent (seven of 29) had 50 percent, and 3 percent (one of 29) had 25 percent concordance. In summary, greater than 70 percent of the patient image sets could be sorted with at least 75 percent concordance by residents. In the senior surgeons' study, 90 percent of patients could be identified as falling into two of five possible groups. Senior raters demonstrated nearly 70 percent repeatability between sortings. CONCLUSION: These findings support the hypothesis that there are identifiable and reproducible patterns of varying calvarial dysmorphology in patients with sagittal craniosynostosis. PMID- 17415250 TI - Surgical treatment of periocular hemangiomas: a single-center experience. AB - BACKGROUND: At Great Ormond Street Hospital for Children, patients with periocular hemangiomas are assessed in a multidisciplinary team setting using a protocol developed in 1999. As part of this protocol, surgery is indicated for lesions that continue to cause amblyopia despite treatment with systemic or intralesional steroids or both. Surgery is performed by one of the authors (D.D.). This experience is described. METHODS: A retrospective notes review was undertaken of surgically excised periocular hemangiomas. RESULTS: Eighteen consecutive cases (15 girls and three boys) were identified as having surgical treatment over a 5-year period. The mean age at the time of surgery was 22 months (range, 5 months to 3 years). All but one of the patients had amblyopia or a threat to normal visual development. The follow-up ranged from 1 month to 4 years. Three were lower lid lesions and 15 were upper lid lesions. When there was a threat to normal visual development, preoperative imaging by ultrasound, magnetic resonance imaging, or contrast computed tomography was performed. No rebound growth or deterioration in visual development occurred in this group of patients. CONCLUSIONS: There is limited experience in the literature of excisional surgery for the treatment of periocular hemangiomas. The authors present a series of 18 patients who have been treated at Great Ormond Street Hospital for Children after either failed medical therapies or after patient request. With the appropriate indications and surgical principles, periocular hemangiomas in children can be safely and effectively excised. PMID- 17415251 TI - Limited incision nonendoscopic brow lift. AB - BACKGROUND: The authors compared the nonendoscopic brow lift technique to the popular endoscopic procedure to determine whether it offers a less complex and less expensive but equally effective alternative. METHODS: A retrospective comparison of the senior author's experience with the endoscopic brow lift (100 patients; years 1999 to 2004) and the nonendoscopic brow lift (93 patients; years 2002 to 2005) was conducted. Using a three-incision approach for both procedures (one midline and two temporal), endoscopic visualization was used to assist in the last 2 cm of subperiosteal dissection over the superior orbital rim only in the endoscopic technique. In the nonendoscopic technique, this final dissection was performed without the endoscope, and the expected path of the supraorbital and supratrochlear neurovascular bundles through preoperative marking of their meridians was respected. Effective brow elevation, operative times, size of incisions, complications, and overall patient satisfaction were compared between groups. RESULTS: The authors found no significant difference in average brow elevation between the two brow lift groups (4 mm). However, the nonendoscopic brow lift was completed, on average, 20 minutes faster than the endoscopic brow lift (30 minutes versus 50 minutes) and required a smaller incision than the endoscopic brow lift (2 cm versus 2.5 cm). No nonendoscopic patient experienced permanent complications, but one endoscopic patient developed permanent paresthesias of the forehead secondary to supraorbital/supratrochlear nerve injury. Overall patient satisfaction was equivalent in both groups. CONCLUSIONS: The limited incision nonendoscopic brow lift is a safe and effective alternative to the endoscopic technique. With thorough anatomical knowledge of this region, it offers equivalent brow elevation, shorter operative times, smaller incisions, similarly low complications rates, and patient satisfaction and eliminates the need for costly and cumbersome endoscopic equipment. PMID- 17415252 TI - An experimental model for improving fat graft viability and purity. AB - BACKGROUND: Autologous fat is an excellent soft-tissue filler, given its abundance, ease of harvest, and natural appearance. However, graft longevity is unpredictable and is reported in the literature to be between 3 months and 8 years. METHODS: A genetically identical, age- and sex-matched mouse experiment was used to develop a model. Inguinal fat pads were subjected to different harvest and preparatory techniques. Primary endpoints-viability and purity-were assessed with the trypan blue viability assay and component counting with a hemocytometer. RESULTS: Viability and purity were highest after excisional harvest versus blunt or needle harvest, presumably secondary to differences in cellular trauma. Saline wash or centrifugation after harvest produced modest but statistically significant improvements in viability and purity. However, if grafts harvested in any fashion were treated with an initial collagenase digestion followed by an idealized centrifugation regimen and a single wash step, viability and purity were consistently 96 percent and 93 percent, respectively. CONCLUSIONS: Using an in vitro murine model, the authors have systematically developed a clinically practical model for creating a pure single-cell suspension of viable adipocytes that is reproducible, regardless of tissue harvest method. PMID- 17415253 TI - Spontaneous breast enlargement following liposuction of the abdominal wall: does a link exist? AB - BACKGROUND: A retrospective study was undertaken to determine the specific incidence of breast enlargement following liposuction of the abdomen (alone or in combination with the flanks), and to compare its effect with a control group of patients who had undergone abdominoplasty only and, where possible, identify corresponding variables. METHODS: Forty-eight of 84 patients (57 percent) who had undergone a tumescent liposuction procedure of at least the abdominal wall and/or flanks and 53 of 104 patients (51 percent) who had undergone abdominoplasty met the entry criteria and formed the study group and the control group, respectively. The medical records were reviewed retrospectively, patient interviews were conducted and, where possible, the patients were examined. Patients who had undergone previous breast or abdominal wall surgery or who had a history of or were breast-feeding at the time of the study were excluded. RESULTS: In the liposuction group, 23 of the 48 patients (48 percent) reported an increase in their breast size postoperatively. This could objectively be confirmed (by an actual increase of at least one bra cup size) in 19 patients (40 percent). Nine of 19 responders (47 percent) presented with a weight gain of as little as 4 percent of body mass index following liposuction, whereas this was observed in only one of 29 of the nonresponder group (3 percent) (p < 0.0001). In the abdominoplasty group, 11 patients (21 percent) claimed to have perceived an increase in breast size, which was objectively confirmed by an increase in bra cup size in six (11 percent) only. Four of the six responders (with an increased cup size) reported a weight gain from as little as 4 percent of body mass index, compared with nine of the remaining 47 patients (19 percent) comprising the nonresponder group (p < 0.01). CONCLUSIONS: Liposuction of the abdominal wall and/or flanks is followed by breast enlargement in a significant number of patients (40 percent), a risk that is significantly higher when compared with patients who have undergone abdominoplasty only. Patients should be informed about the possibility and risk of breast enlargement following liposuction of the abdominal wall in particular. PMID- 17415254 TI - Factors impacting thromboembolism after bariatric body contouring surgery. AB - BACKGROUND: The purpose of this study was to define the risk of venous thromboembolism within the massive weight loss population undergoing body contouring procedures. METHODS: Retrospective analysis of massive weight loss patients who had body contouring operations between March of 1998 and September of 2004 was performed. Patient factors studied included age, gender, medical comorbidities including history of thromboembolic complications, depression, tobacco use, preoperative/postoperative body mass index, surgery, and transfusion. RESULTS: There were 138 cases, and the female-to-male ratio was 5:1. Procedures were often combined: 128 patients had abdominal surgery, 36 had a back lift, 41 had brachioplasty, 29 had chest surgery, and 47 had a thigh lift. The most common complications were related to healing (n = 28) and seroma (n = 18). Three patients had postoperative deep venous thrombosis requiring anticoagulation, and one had a fatal pulmonary embolism, making the overall venous thromboembolism risk 2.9 percent. The mean body mass index at contour was 48.5 for patients with venous thromboembolism versus 31.8 for patients who did not develop venous thromboembolism (p = 0.01). Looking at this subgroup of 45 patients, the risk of venous thromboembolism was 8.9 percent, with no risk found in patients with a body mass index less than 35 (p = 0.01). CONCLUSIONS: The risk of venous thromboembolism with contouring surgery for massive weight loss is comparable to that for gastric bypass surgery. Body mass index in the obese range appears to be a leading risk factor. The authors' data support routine prophylaxis against venous thromboembolism. Recommendations for high-risk patients are discussed. PMID- 17415256 TI - Flap ischemia after pedicled tissue transfer to a lateral thigh wound: an example of complication assessment using the end result system of E. A. Codman. PMID- 17415258 TI - Ernest Amory Codman, M.D. (1869 to 1940): the influence of the End Result Idea on plastic and reconstructive surgery. PMID- 17415260 TI - About teaching. PMID- 17415263 TI - Use of prelaminated free forearm flap with tissue expansion in reconstruction. PMID- 17415264 TI - Liposuction and pulmonary embolism: the role of D-dimers. PMID- 17415265 TI - On the ethics of composite tissue allotransplantation (facial transplantation). PMID- 17415267 TI - Radix malorum cupiditas est. PMID- 17415268 TI - Improvised instrument for expander placement. PMID- 17415270 TI - Anthropometric analysis of the slant of palpebral fissures. PMID- 17415271 TI - Fatal side effect of maggot treatment on wound healing. PMID- 17415272 TI - Use of the internal mammary vessels in breast reconstruction: a cautionary note. PMID- 17415273 TI - A simple method of preventing snapper fallout during syringe liposuction. PMID- 17415274 TI - Primary repair of zone II flexor tendon lacerations at 5 weeks and 5 days after injury. PMID- 17415275 TI - A technique to facilitate accurate and atraumatic tendon repair. PMID- 17415276 TI - Management of an unusual extreme extension contracture of an index finger: a custom-designed exercise program in achieving a good range of movement and prevention of recontraction with the reverse fasciocutaneous forearm flap and full-thickness skin graft. PMID- 17415277 TI - Uncommon features of an uncommon problem: embedded ring syndrome in a child. PMID- 17415278 TI - Botulinum toxin as a treatment for leg ulcers. PMID- 17415279 TI - A custom-made latex conduit for simulation of microvascular repair. PMID- 17415280 TI - Adverse reactions to wound dressings: a financial incendiary. PMID- 17415282 TI - Aliskiren (Tekturna) for hypertension. PMID- 17415283 TI - Subcutaneous immune globulin (SCIG). PMID- 17415284 TI - What can doctors do about health literacy? PMID- 17415285 TI - A tale of two fungi in a person with HIV. PMID- 17415286 TI - Kinesigenic paroxysmal hemidyskinesia as the initial presentation of multiple sclerosis. AB - Fewer than a hundred cases of paroxysmal dystonia have been described in patients with multiple sclerosis (MS). Even fewer cases of hemidyskinesia triggered by repetitive movements (paroxysmal kinesigenic hemidyskinesia--PKD) have been reported in MS patients. We describe the case of a woman, age 18 years at the onset of MS, and a man age 35 years at the onset of MS, who presented with PKD as the initial symptom. Magnetic resonance images (MRI) of these patients showed different areas of acute lesions possibly related to PKD; MRI of one of the patients demonstrated 1 lesion in the subcortical parietal area and another in the thalamic region and showed 2 lesions in the cervical spinal cord in the other patient. PMID- 17415287 TI - Comparing the efficacy of medications for ADHD using meta-analysis. AB - OBJECTIVE: Medications used to treat attention-deficit/hyperactivity disorder (ADHD) have been well researched, but comparisons among drugs are hindered by the absence of direct comparative trials. METHODS: We analyzed recent published literature on the pharmacotherapy of ADHD to describe the variability of drug placebo effect sizes. A literature search was conducted to identify double-blind, placebo-controlled studies of ADHD youth published after 1979. Meta-analysis regression assessed the influence of medication type and study design features on medication effects. RESULTS: Twenty-nine trials met criteria and were included in this meta-analysis. These trials studied 15 drugs using 17 different outcome measures of hyperactive, inattentive, impulsive, or oppositional behavior. The most commonly identified treatments included both methylphenidate and amphetamine compounds. After stratifying trials on the class of drug studied (short-acting stimulant vs long-acting stimulant vs nonstimulant), we found significant drug differences for both study design variables and effect sizes. The differences among the 3 classes of drug remained significant after correcting for study design variables. CONCLUSION: Uniformity appears to be lacking in how medication effectiveness is assessed and in many study design parameters. Comparing medication effect sizes from different studies will be biased without accounting for variability in study design parameters. Although these differences obscure comparisons among specific medications, they do allow for conclusions about the differential effects of broad classes of medications used to treat ADHD. PMID- 17415288 TI - Advanced practice nurses say "No" to a mandatory Doctor of Nursing Practice degree. PMID- 17415289 TI - Endocannabinoids--the brain's own marijuana--may be linked to the metabolic syndrome. PMID- 17415290 TI - A safety assessment of topical calcineurin inhibitors in the treatment of atopic dermatitis. AB - Atopic dermatitis (AD) is a common childhood disease that can disrupt the lives of patients and their families and, in turn, affect their quality of life. The goal of treatment is the long-term control of AD by minimizing the frequency and severity of flares. Topical corticosteroids of various potencies have been the mainstay of pharmacologic treatment of AD flares. In the past few years, the introduction of topical calcineurin inhibitors (TCIs) has provided physicians with an effective, well-tolerated alternative to topical corticosteroids. In January 2006, a boxed warning and a patient medication guide were added to TCI product labels in the United States after the US Food and Drug Administration raised concerns about their safety. These concerns were based on rare cases of skin malignancy and lymphoma, and a theoretical risk stemming from the systemic use of calcineurin inhibitors in animal studies and transplant patients. However, the boxed warning states that no causal link has been established between TCI use and malignancy. Pharmacokinetic studies have also shown that treatment with TCIs leads to only minimal systemic absorption. In addition, controlled, blinded studies have found no evidence of systemic immunosuppression and no causal relationship between the use of TCIs and the occurrence of lymphoma or other malignancies. Overall, TCIs have been shown to be an effective and valuable treatment option for AD. PMID- 17415291 TI - Laboratory results that should be ignored. PMID- 17415292 TI - A fatal case of propylthiouracil-induced ANCA-positive vasculitis. AB - A 60-year-old man with a history of Graves' disease, treated with propylthiouracil (PTU), presented with a rash, pancytopenia, and lymphadenopathy. The patient subsequently developed acute renal failure and diffuse alveolar hemorrhage. Despite discontinuation of PTU and aggressive therapy including high dose steroids, cyclophosphamide, and plasma exchange, the patient died. PTU induced antineutrophilic cytoplasmic antibody (ANCA)-positive vasculitis has been well described in the literature. Patients with this condition are almost always positive for pANCA or anti-myeloperoxidase (MPO). Patients can have varying presentations, and symptoms usually resolve with discontinuation of the drug. Some patients, however, require high-dose steroids, immunosuppressives, or plasmapheresis. Rarely, fatalities can occur from this condition. The mechanism of PTU-induced vasculitis is not well understood. PMID- 17415293 TI - Brief communication: economic comparison of opportunistic infection management with antiretroviral treatment in people living with HIV/AIDS presenting at an NGO clinic in Bangalore, India. AB - CONTEXT: Highly active antiretroviral treatment (HAART) usage in India is escalating. With the government of India launching the free HAART rollout as part of the "3 by 5" initiative, many people living with HIV/AIDS (PLHA) have been able to gain access to HAART medications. Currently, the national HAART centers are located in a few district hospitals (in the high- and medium-prevalence states) and have very stringent criteria for enrolling PLHA. Patients who do not fit these criteria or patients who are too ill to undergo the prolonged wait at the government hospitals avail themselves of nongovernment organization (NGO) services in order to take HAART medications. In addition, the government program has not yet started providing second-line HAART (protease inhibitors). Hence, even with the free HAART rollout, NGOs with the expertise to provide HAART continue to look for funding opportunities and other innovative ways of making HAART available to PLHA. Currently, no study from Indian NGOs has compared the direct and indirect costs of solely managing opportunistic infections (OIs) vs HAART. OBJECTIVE: Compare direct medical costs (DMC) and nonmedical costs (NMC) with 2005 values accrued by the NGO and PLHA, respectively, for either HAART or exclusive OI management. STUDY DESIGN: Retrospective case study comparison. SETTING: Low-cost community care and support center--Freedom Foundation (NGO, Bangalore, south India). PATIENTS: Retrospective analysis data on PLHA accessing treatment at Freedom Foundation between January 1, 2003 and January 1, 2005. The HAART arm included case records of PLHA who initiated HAART at the center, had frequent follow-up, and were between 18 and 55 years of age. The OI arm included records of PLHA who were also frequently followed up, who were in the same age range, who had CD4+ cell counts < 200/microliter (mcL) or an AIDS-defining illness, and who were not on HAART (solely for socioeconomic reasons). A total of 50 records were analyzed. Expenditures on medication, hospitalization, diagnostics, and NMC (such as food and travel for a caregiver) were calculated for each group. RESULTS: At 2005 costs, the median DMC plus NMC in the OI group was 21,335 Indian rupees (Rs) (mean Rs 24,277/-) per patient per year (pppy) (US $474). In the HAART group, the median DMC plus NMC was Rs 18,976/- (mean Rs 21,416/-) pppy (US $421). Median DMC plus NMC pppy in the OI arm was Rs 13623.7/- paid by NGO and Rs 1155/- paid by PLHA. Median DMC and NMC pppy in the HAART arm were Rs 1425/- paid by NGO and Rs 17,606/- paid by PLHA. CONCLUSIONS: Good health at no increased expenditure justifies providing PLHA with HAART even in NGO settings. PMID- 17415294 TI - Pandemic influenza preparedness. PMID- 17415295 TI - Obscurity vs transparency in health measurement. PMID- 17415296 TI - Public reason and bioethics. PMID- 17415297 TI - Herniation of gravid uterus: report of 2 cases and review of literature. AB - Anterior abdominal wall hernias are uncommon, and herniation of a gravid uterus into these hernias is even rarer. Although reducible initially, the herniation of pregnant uterus may be complicated by incarceration and subsequent strangulation within the hernial sac, late in the course of pregnancy. There is no consensus over the management of this rare condition. Each case should be individualized. If uncomplicated, a conservative approach until term followed by delivery and herniorrhaphy is a good option. Here, 2 cases of herniation of gravid uterus into the anterior abdominal wall are described along with a brief review of literature pertaining to its presentation, complications, and management. PMID- 17415298 TI - A 14-year-old girl with abdominal pain and emesis. PMID- 17415299 TI - The patient safety movement finally is saving lives and raising hopes. PMID- 17415300 TI - Blueprint for change: will new hospitals be safer hospitals? PMID- 17415301 TI - Is medical student choice of a primary care residency influenced by debt? AB - CONTEXT: The average medical student accumulates more than $120,000 in debt upon graduation. OBJECTIVE: The purpose of this study was to investigate whether medical student debt affects residency choice. DESIGN: This was a cross-sectional research study. SETTING: This study was a 5-year analysis of student debt and residency choice for 2001-2005 graduates from 3 US medical schools (n = 2022): Tulane University School of Medicine, New Orleans, Louisiana; University of South Florida College of Medicine, Tampa, Florida; and Louisiana State University School of Medicine in New Orleans. MAIN OUTCOME MEASURES: Individual student data were collected from offices of financial aid for debt prior to and during medical school to determine total debt at graduation. Total debt (adjusted for inflation) was compared with residency match results coded according to specialties listed in the Graduate Medical Education Directory 2005-2006. Graduates were coded into either primary care (PC) or nonprimary care (NPC) specialty categories. Logistic regression for the choice of a PC residency was used with 4 predictors: (1) total debt, (2) medical school, (3) year of graduation, and (4) number of years of training required for a residency program. RESULTS: Mean total debt for the study population was $89,807 (SD = 54,925). Graduates entering PC did not have significantly less total debt than those entering NPC ($87,206 vs $91,430; P = .09). Further, total debt was not a predictor of a PC residency after adjusting for medical school, year of graduation, and years of training in residency (P = .64). CONCLUSION: There is no association between PC residency choice and debt. We conclude that medical students make residency decisions on the basis of a complex set of factors. PMID- 17415302 TI - Repetitive prostatic massage and drug therapy as an alternative to transurethral resection of the prostate. AB - We describe 5 men with urinary retention and indwelling urethral catheters who were treated with repetitive prostatic massage, antimicrobials, alpha blockers, and--in 2 cases--finasteride. We retrospectively reviewed the charts of all patients presenting to the genitourinary clinic with indwelling urinary catheters during a 1-year period. Five men (mean age, 70 years; range, 64-76; SD 4.47) presented to the Manila Genitourinary Clinic (Cebu Branch), Cebu, Philippines, wearing indwelling urinary catheters placed for acute urinary retention. Urologists had told all 5 men that they needed to undergo transurethral resection of the prostate (TURP). The Cebu genitourinary physician removed the catheters, instituted repetitive prostatic massage, and diagnosed all 5 patients with prostatitis. All 5 patients received repetitive prostatic massage, alpha-blocker medication, and antibiotic therapy, whereas finasteride was given to 2 patients. During treatment, statistically significant improvements occurred in global symptom severity scores, urethral white blood cell (WBC) counts, WBC counts of the expressed prostatic secretions (EPS), EPS red blood cell (RBC) counts, urinary WBC counts, and urinary RBC counts. Fluorescing Chlamydia elementary bodies disappeared in 3 of the 4 positive patients by the end of treatment. (One patient was not available for retesting.) Repetitive prostatic massage, antimicrobial therapy, alpha-blocker therapy, and--in 2 cases--finasteride enabled catheter removal in all 5 men (100%) as well as successful urination in all 5 men (100%). TURP has been prevented for a mean of 2.53 years (range, 16-38 months). PMID- 17415304 TI - Twenty-five years of AIDS: where are we now? PMID- 17415305 TI - Financial conflicts of interest are related to voting patterns at FDA Advisory Committee meetings. PMID- 17415306 TI - Case 14: a woman with bilateral Bell's palsy. PMID- 17415307 TI - Esophageal duplication cyst--a guest case in robotic and computer-assisted surgery from the University of Nebraska Medical Center. PMID- 17415308 TI - Will transparency of performance information improve quality? PMID- 17415309 TI - Another hidden epidemic: limited health literacy. PMID- 17415310 TI - Prevalence of coronary risk factors, clinical presentation, and complications in acute coronary syndrome patients living at high vs low altitudes in Yemen. AB - BACKGROUND: A comparative retrospective study was performed to compare the distribution of risk factors and complications in patients with acute coronary syndrome (ACS) at high-altitude vs low-altitude areas in Yemen. METHODS: The records of 768 patients from Sana'a (high altitude) and Aden (low altitude) were reviewed. Risk factors assessed were age, hypertension, diabetes mellitus, hyperlipidemia, cigarette smoking, and reported history and family history of coronary artery disease (CAD). Complications of ACS of interest were heart failure, arrhythmias, cerebrovascular accident (CVA), and death. RESULTS: The mean age of ACS patients at high altitude was significantly lower than those at low altitude (55.3 years [SD = 8.2] vs 56.8 years [SD = 7.1]; P = .007). History of hyperlipidemia was significantly higher in high-altitude patients than in low altitude patients (49.2% vs 38.3%; odds ratio [OR] = 1.563; P = .002). Reported history of CAD was also significantly higher at higher altitudes (16.7% vs 9.4%; OR = 1.933; P = .003). Previous history of diabetes mellitus and tobacco smoking was slightly higher with borderline significance. Hypertension and reported family history of CAD were comparable among high- and low-altitude patients. In terms of in-hospital complications, CVAs were significantly higher in high altitude patients than in low-altitude patients (7.8% vs 4.4%; P = .0001). Heart failure, arrhythmias, and death rates were comparable in both groups of patients. Wall motion abnormalities were comparable, whereas the ejection fraction was lower in the high-altitude patients (49.8% [SD = 16.08] vs 54.8% [SD = 16.23]; P = .0001). CONCLUSIONS: ACS occurs at a younger age at high altitudes. Patients who live in high-altitude regions are also more likely to have hyperlipidemia and a previous history of CAD. Stroke and reduced left ventricular ejection fraction (LVEF) occur more commonly in high-altitude ACS patients. High altitude may generally be a risk factor for ACS. PMID- 17415311 TI - Pulmonary hypertension, hyperthyroidism, and fenfluramine: a case report and review. AB - We report a case of pulmonary hypertension presenting with sudden cardiac arrest, hyperthyroidism and fenfluramine usage. This fatal case of pulmonary hypertension and valvular heart disease is associated with the use of an anorectic drug that had been withdrawn from the market more than eight years ago. Clinicians should alert to the side effects of appetite suppressant and slimming agents. The association between pulmonary hypertension in relation to hyperthyroidism and fenfluramine usage is reviewed. PMID- 17415312 TI - Understanding the scourge of HIV/AIDS in sub-Saharan Africa. AB - Sub-Saharan Africa is the part of the world that has been hit hardest by the HIV epidemic. To fight the spread of HIV in the continent, it is necessary to know and effectively address the factors that drive the spread of HIV. The purpose of this article is to review the factors associated with the spread of the HIV epidemic in sub-Saharan Africa and to propose 6 essential activities, which we refer to by the acronym "ESCAPER," to help curb the spread of HIV/AIDS in Africa. PMID- 17415313 TI - We can do more to avoid medication errors. PMID- 17415314 TI - Transparent measures are essential for coordinating and rewarding quality improvement in healthcare. PMID- 17415316 TI - Addition of basal insulin to oral antidiabetic agents: a goal-directed approach to type 2 diabetes therapy. AB - The objective of this article is to review current findings in the published literature on the efficacy of insulin therapy in combination with oral antidiabetic agents, with a focus on practical information that might help to provide an evidence-based template for selecting how best to combine oral agents and basal insulin in patients with type 2 diabetes. Here we review the current oral agents used to treat type 2 diabetes, their mechanisms of action, and how they can be combined with insulin therapy to help patients achieve guideline recommended glycemic goals. While practical advice exists for initiating a therapeutic regimen comprised of basal insulin and oral agent(s), direction as to appropriate therapy for individual patients with differing physiologic requirements is needed. Oral antidiabetic therapy in combination with insulin provides an effective therapeutic option for patients who are unable to achieve or maintain glycemic goals on oral therapy alone. PMID- 17415315 TI - Biological response modifiers in cancer. AB - We have seen a surge in the use of immunotherapy for the treatment of cancer. Biological response modifiers can act passively by enhancing the immunologic response to tumor cells or actively by altering the differentiation/growth of tumor cells. Active immunotherapy with cytokines such as interferons (IFNs) and interleukins (IL-2) is a form of nonspecific active immune stimulation. The use of IL-2 has recently been approved by the United States Food and Drug Administration (FDA) for the treatment of renal cell carcinoma and metastatic colorectal cancer. Considerable success has been achieved with the use of immunotherapy, especially in the area of passive immunotherapy using monoclonal antibodies--in particular, radiolabeled monoclonal antibodies. In addition to the various monoclonal antibodies that have been used in clinical trials, other strategies such as the use of antiangiogenic agents and matrix metalloprotease inhibitors (MMPIs) have also met with some success. Recently, the FDA approved bevacizumab, an anti-vascular endothelial growth factor (VEGF) agent, for the treatment of metastatic melanoma. This review also sheds light on the various angiogenesis inhibitors in clinical trials, the increasing use of thalidomide in cancer, and the upcoming potential cancer vaccines designed to activate cell mediated immune responses against tumor antigens. PMID- 17415317 TI - A survey of neurologists on bothersome patient behaviors. AB - CONTEXT: Although much has been published on patient complaints about physicians, little is known about patient behaviors that physicians find bothersome. OBJECTIVE: To obtain more information about bothersome patient behaviors, we performed a pilot survey of neurologists. METHODS: A survey was performed among neurologists attending the Texas Neurological Society Winter Conference in 2006 in Austin, Texas. Using 5-point Likert scales, respondents rated 30 patient behaviors on how often they observed or experienced the behavior in the last year and how bothersome the behavior was to them. Respondents were asked to list their top 5 most bothersome patient behaviors, provide examples of unnecessary phone calls received after hours, and provide any other comments about the survey. RESULTS: The response rate was 42.3% (78 respondents). Almost all of the patient behaviors were found to be bothersome or very bothersome. The top 5 most bothersome patient behaviors from most to least were the following: no show for appointment, verbally abusive with your staff, poor compliance with medications or treatment, late for appointment, and do not know the medications that they are taking. Of the 30 items, those behaviors as well as answering cell phones during office visits and unnecessary phone calls after hours were among the most highly rated as bothersome. CONCLUSIONS: Some of the bothersome patient behaviors may have deleterious effects on patient care, whereas others may make the practice of medicine unpleasant for physicians. A number of the behaviors may be reduced by providing patients with practice policies. Others are best addressed through ongoing communication and education. PMID- 17415318 TI - Primary care physicians: a vanishing species or a profession reinvented? PMID- 17415319 TI - Is the food industry playing games with our children? PMID- 17415320 TI - Effects of an 8-week high-protein or high-carbohydrate diet in adults with hyperinsulinemia. AB - CONTEXT: Incidence of insulin resistance (IR) in Americans is steadily rising. IR may be ameliorated with < or = 5% loss in body weight. OBJECTIVE: To examine effects of 2 weight-loss diets on body weight and composition in overweight adults with IR. DESIGN: Participants randomly assigned to a high-protein, low-fat (HPLF) or a high-carbohydrate, low-fat (HCLF) diet for 8 weeks. SETTING: All meals prepared and weighed in the metabolic kitchen at Arizona State University. Lunch consumed on-site; all other meals packaged for home consumption. PATIENTS: Twenty overweight, healthy participants with elevated fasting serum insulin (> or = 15 microU/L) were recruited. INTERVENTIONS: Both diets were low-fat (27% kcal from fat; < 7% saturated, < or = 10% monounsaturated, and < or = 10% polyunsaturated) and energy-restricted (energy levels were 1200, 1500, 1700 or 2000 kcal); HPLF: 32% protein, 41% carbohydrate; HCLF: 59% carbohydrate, 14% protein. Energy levels were assigned on the basis of participant's resting metabolic rate. MAIN OUTCOME MEASURES: Body composition, etabolic indices, fasting plasma glucose, and insulin. RESULTS: No significant differences were found in the main outcome measures between the diets. Body weight (HPLF: -4.9 kg; HCLF: -4.0 kg) and total percent body fat (HPLF: -1.5%; HCLF: -0.4%) significantly reduced from baseline to week 8 (P = .005 and P = .035, respectively). CONCLUSION: Both diets promoted > or = 5% loss in body weight and significantly reduced percent body fat. PMID- 17415321 TI - End the war on obesity: make peace with your patients. PMID- 17415322 TI - Ingestion of superwarfarin leading to coagulopathy: a case report and review of the literature. AB - Superwarfarins are found in many pesticides, including D-con, Prufe I and II, Ramik, Talon-G, Ratak, and Contrac. Ingestion of can lead to significant morbidity and even mortality. Physicians need to consider this diagnosis in any patient presenting with coagulopathy of unclear etiology. We present a patient with superwarfarin-induced coagulopathy and review previous cases in adults in the literature. The patient is a 60-year-old man who presented to our medical center with painless hematuria. Laboratory studies revealed an elevated prothrombin time (PT) (42.5 seconds), partial thromboplastin time (PTT) (64.6 seconds), and international normalized ratio (INR) of 7. Liver-associated enzymes were normal, and complete blood cell count (CBC) showed no evidence of disseminated intravascular coagulation. Subsequent work-up included the absence of an inhibitor by mixing study and deficiencies of vitamin K-dependent coagulation factors. The patient's warfarin level was negative. A brodifacoum level was positive, confirming superwarfarin-induced coagulopathy. The patient is currently doing well with normal coagulation studies after receiving high doses of vitamin K for several weeks. The cause of his exposure to superwarfarin remains uncertain. Physicians need to be cognizant of this unusual cause of coagulopathy in adults. The appropriate diagnostic work-up and unique features of therapy are discussed. PMID- 17415323 TI - A comparative study of HIV/AIDS: the knowledge, attitudes, and risk behaviors of schizophrenic and diabetic patients in regard to HIV/AIDS in Nigeria. AB - CONTEXT: Studies on knowledge and risk behaviors related to HIV/AIDS reported from developed countries have shown that people with psychiatric disorders constitute a special risk group. In Nigeria, although similar studies have been conducted on various population groups, there has, so far, been no reported study on people suffering from psychiatric disorders. OBJECTIVE: The present study set out to compare knowledge, attitudes, and risk behaviors related to HIV/AIDS among schizophrenic patients and diabetic patients. METHOD: Ninety-eight consecutive schizophrenic patients attending the outpatient clinics of a psychiatric hospital over a period of 8 weeks completed an interviewer's administered questionnaire. The interview covered demographics, risk behaviors, knowledge related to HIV/AIDS, and patients' attitudes toward people infected with HIV/AIDS. Their responses were compared with those of 56 diabetic patients who were similarly interviewed in a teaching hospital. RESULTS: Compared with the diabetic patients, the schizophrenic patients were significantly less sexually active in the previous 12 months (P < .05). They had more misconceptions about HIV/AIDS and were less tolerant towards people living with HIV/AIDS compared with the diabetic patients. They were also more likely to engage in high-risk behaviors. CONCLUSION: Mental health providers rarely educate psychiatric patients about HIV/AIDS and should be more involved in doing so. Despite being less sexually active, patients with schizophrenia engaged in risk behaviors as did the diabetic patients. PMID- 17415324 TI - Cutaneous and presumed visceral leishmaniasis in a soldier deployed to Afghanistan. AB - Leishmaniasis has been frequently diagnosed in US military personnel returning from duty in Southwest Asia. The majority of cases have demonstrated cutaneous disease, although a few cases of visceral disease have been documented. We present the case of an immunocompetent, HIV-negative, US Army soldier who suffered both visceral and cutaneous manifestations of leishmaniasis after returning from deployment in Afghanistan. Overlap of cutaneous and visceral involvement is rare and has not been reported in our cohort. Latent Plasmodium vivax infection may have been an exacerbating cofactor. We discuss this case and present a review of the literature. PMID- 17415325 TI - Awakening Hippocrates: a call for health providers to serve where most needed. PMID- 17415326 TI - Nuclear power is contraindicated as a solution to global warming because of nuclear mutagenesis. PMID- 17415327 TI - Deaths of detainees in the custody of US forces in Iraq and Afghanistan from 2002 to 2005. AB - In light of the large number of detainees who continue to be taken and held in US custody in settings with limited judicial or public oversight, deaths of detainees warrant scrutiny. We have undertaken the task of reviewing all known detainee deaths between 2002 and early 2005 based on reports available in the public domain. Using documents obtained from the Department of Defense through a Freedom of Information Act request, combined with a review of anecdotal published press accounts, 112 cases of death of detainees in United States custody (105 in Iraq, 7 in Afghanistan) during the period from 2002 to early 2005 were identified. Homicide accounted for the largest number of deaths (43) followed by enemy mortar attacks against the detention facility (36). Deaths attributed to natural causes numbered 20. Nine were listed as unknown cause of death, and 4 were reported as accidental or natural. A clustering of 8 deaths ascribed to natural causes in Iraq in August 2003 raises questions about the adequacy and availability of medical care, as well as other conditions of confinement that may have had an impact on the mortality rate. PMID- 17415328 TI - Brain-stem listeriosis: a comparison of SPECT and MRI findings. AB - Listeria monocytogenes, although uncommon as a cause of illness in the general population, can result in serious illness when it affects pregnant women, neonates, the elderly, and immunocompromised individuals. Typically, it is a food borne organism. This report describes a case of brain-stem listeriosis in a previously healthy 51-year-old woman. The diagnosis was based on clinical findings, the results of cerebrospinal fluid (CSF) analysis, CSF culture, and magnetic resonance imaging (MRI) findings. MRI demonstrated upper brain stem and cerebellar peduncle involvement. In addition, Tc-99m exametazime (HMPAO)-labeled single photon emission computed tomography (SPECT) of the brain revealed bilateral cerebellar hypoperfusion. Antibiotic therapy resulted in partial clinical recovery after 3 weeks. At the end of 6 months, brain-stem findings had nearly resolved. However, although minimal residual findings were observed on MRI at 6 months, bilateral diffuse cerebellar hypoperfusion remained on Tc-99m HMPAO brain SPECT. PMID- 17415329 TI - Bacterial pericarditis and tamponade due to nonencapsulated Haemophilus influenzae complicating a case of adult community-acquired pneumonia. AB - We report a case of bacterial pericarditis in an immunologically competent adult female caused by nonencapsulated Haemophilus influenzae (H influenzae) that was complicated by the acute development of life-threatening pericardial tamponade. H influenzae is a gram-negative coccobacillus, a pathogen most frequently associated with childhood exanthema (otitis media, meningitis) and, less frequently, adult pneumonia. Encapsulated, type b, or typable H influenzae is the strain implicated in childhood infections. On the other hand, nonencapsulated or nontypable H influenzae is the specific strain most often associated with exacerbation of chronic obstructive airway disease. Bacterial pericarditis caused by either subtype of H influenzae is exceedingly rare. We have located only 15 previously reported cases of H influenzae pericarditis occurring in adults in the world medical literature, the majority of which date back to the pre-antibiotic era. In 12 of these 15 cases (the only cases in which typing could be accomplished), the encapsulated strain of H influenzae was cultured from the pericardial fluid. Thus, to the best of our knowledge, we are reporting here the first case of bacterial pericarditis caused by nonencapsulated H influenzae in an immunologically competent adult. PMID- 17415330 TI - The functioning brains of us physicians should be guidance systems, not books of knowledge. PMID- 17415331 TI - How to create a new consensus on quality improvement measures in health care. PMID- 17415332 TI - Awareness of authorship criteria and conflict: survey in a medical institution in India. AB - CONTEXT: Assignment of appropriate authorship on a scientific paper is important for researchers and the public; inappropriate authorship may generate conflict. OBJECTIVES: To determine the extent of: (a) awareness of criteria for authorship and acknowledgment among researchers in a medical institution, (b) conflict concerning authorship issues in the research environment, and (c) their interrelationship. DESIGN AND SETTING: A questionnaire self-administered by the faculty in a teaching hospital in India in March 2006. MAIN OUTCOME MEASURES: Rate and level of awareness of International Committee of Medical Journal Editors (ICMJE) Uniform Requirements for Manuscripts Submitted to Biomedical Journals, or other rate and level of awareness of international published authorship criteria; rate and type of authorship conflict. RESULTS: Ninety-five teaching faculty were contacted; 77 responded. Fifty respondents (65%) reported that they were aware that authorship criteria existed, but only 34 (44%) correctly identified a source. Overall, 30 (39%) respondents reported conflict over authorship issues relating mainly to ownership of data, gift authorship, and issues like academic competition, personality differences, intellectual passion, or order of authorship. There was no difference between number of authorship criteria correctly identified by respondents who reported conflict compared with those who did not (P = .78). CONCLUSIONS: There was poor awareness of criteria for authorship and acknowledgement among the faculty in research publications. Conflict over authorship issues existed in the research environment, but was not influenced by the level of awareness. PMID- 17415333 TI - Can urine lamivudine be used to monitor antiretroviral treatment adherence? AB - Patient adherence to treatment is an important factor in the effectiveness of antiretroviral regimens. Adherence to treatment could be monitored by estimation of antiretroviral drugs in biological fluids. We aimed to obtain information on the quantity and duration of excretion of lamivudine in urine following oral administration of a single dose of 300 mg and to assess its suitability for adherence monitoring purposes. Spot urine samples were collected before dosing and at 4, 8, 12, 24, 28, 32, 48, 72, and 96 hours post dosing from 10 healthy subjects, and lamivudine was estimated by high-pressure liquid chromatography (HPLC). Lamivudine values were expressed as a ratio of urine creatinine. About 91% of the ingested drug was excreted by 24 hours, and the concentration thereafter in urine was very negligible. A lamivudine value of 0.035 mg/mg creatinine or less at 48 hours is suggestive of a missed dose in the last 24 hours. The study findings showed that estimation of urine lamivudine in spot specimens could be useful in monitoring patient adherence to antiretroviral treatment. However, this needs to be confirmed on a larger sample size and among patients on once-daily and twice-daily treatment regimens. PMID- 17415334 TI - Off-label prescriptions: do we need more restraint or more evidence? PMID- 17415335 TI - Ban trans fats in 2007. PMID- 17415336 TI - Associations between body mass index and the prevalence of low micronutrient levels among US adults. AB - BACKGROUND: Low micronutrient levels observed with increasing adiposity may result from inadequate nutrient intake and/or alterations in nutrient metabolism. OBJECTIVE: To examine the association between body mass index (BMI) and micronutrient levels among a nationally representative sample of US adults aged > or = 19 years. DESIGN: Using nationally representative cross-sectional data from the National Health and Nutrition Examination Survey III (NHANES III), we examined odds ratios of low micronutrient levels using logistic regression adjusting for covariates. MEASUREMENTS: Nutritional biomarker levels (as indicated by serum levels of total carotenoids, alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin, lycopene, vitamin E, vitamin C, selenium, vitamin A, vitamin D, folate, vitamin B12, and red blood cell folate) among men and nonpregnant women, by BMI category. RESULTS: Overweight and obese adults had higher odds of low levels for a number of nutrients compared with normal-weight adults. Odds of being low in multiple micronutrients was most common among overweight and obese premenopausal women. CONCLUSION: These findings underscore the need for further assessment of specific micronutrient inadequacies among persons who are overweight or obese. Specifically, research is needed to determine whether these inadequacies are due to insufficient dietary intake, altered metabolic processes, or both. PMID- 17415337 TI - Low-dose enalapril in the treatment of surgical cutaneous hypertrophic scar and keloid--two case reports and literature review. AB - Hypertrophic scars and keloids are 2 forms of excessive cutaneous scarring that occur in predisposed individuals. The healing process varies greatly among patients, and the risk of a bad scar evolution is unpredictable. Keloids create disfiguring scars with associated erythema and pain or pruritus or restricted range of motion, and are a major cause of morbidity. A fortuitous observation was made by the first author of this study who, at age 54, developed an erythematous and painful postsurgical abdominal keloid scar after undergoing left colectomy for colon adenocarcinoma. Four months later, after treatment with low-dose enalapril (10 mg, once a day) for mild arterial hypertension, her keloid scar rapidly improved and she eventually made a complete recovery. second case involved a 70-year-old female with diabetes who was affected by a long-standing postsurgical abdominal keloid scar of 2 years' duration. She was intentionally treated with the same low dose of enalapril, and, after 6 months of therapy, the bad scar showed marked improvement. We conducted an exhaustive search of the literature pertaining to the wound healing process, specifically to determine whether angiotensin-converting enzyme (ACE) inhibitors have a healing effect on wounds. ACE inhibitors are known to induce reduction of left ventricular collagen content and to attenuate remodeling during the postinfarctual period (thus improving ventricular function), and they have been shown to exert a pulmonary antifibrotic effect. After conducting this literature search, it became apparent that no data on cutaneous scars and ACE inhibitors are available. During the posttraumatic or postoperative stage, it is useful to achieve the best possible aesthetic results and to decrease the risk of a disfiguring keloid scar, thereby avoiding revision surgery; to this purpose, an early treatment with a low dose of enalapril is a possible solution, even if further confirmatory observations are needed. PMID- 17415339 TI - Is America's meth scourge really coming to an end? PMID- 17415340 TI - I use fingernail surgery to remove my seborrheic keratoses. PMID- 17415341 TI - Intrahepatic lymphocyte phenotypes in hepatitis C virus infection: a comparison between cirrhotic and non-cirrhotic livers. AB - AIM: The pathogenesis of viral hepatitis involves the activation of cellular immunity, including intrahepatic lymphocytes (IHL). Lym-phocyte phenotypes play a fundamental role in the pathogenesis of chronic hepatitis C virus (HCV) infection, the progression of liver fibrosis and subsequent hepatocellular carcinoma. The aim of this study was to evaluate the frequency of intrahepatic mononuclear cell phenotypes in patients with chronic HCV. Another aim was to assess the relationship of nonparenchymal cells with liver fibrosis. METHODS: Liver fibrosis was evaluated with the Histologic Activity Index. Fourteen liver biopsies showed mild fibrosis (group 1), and 11 bridging fibrosis (group 2). Fourteen samples were explants from HCV patients who underwent liver transplantation (group 3). CD4 and CD8 T-lymphocytes, CD20 (B lymphocytes), CD16 (macrophage), and CD57 (NK) cells were detected using monoclonal antibodies on paraffin-embedded tissue. RESULTS: A minority of lobular cells stained for T- or B-lymphocytes. Most lobular cells stained with macrophage antibodies, and were more common in bridging fibrosis, compared to mild fibrosis. The percentages of lobular CD4 and CD8 cells were significantly lower in regenerative nodules of cirrhotic livers. There was a strong negative correlation between lobular CD8 and fibrosis score (R= -0.65), and a strong positive correlation between CD16-stained mononuclear cells (macrophages) and fibrosis score (R=0.66). In portal and periportal areas, CD4 but not CD8 lymphocytes decreased in parallel with fibrosis. B-lymphocytes were more commonly found in the portal areas than in the lobule. CD57-positive cells were rare in both lobule and portal areas, and their frequency was not different in the three groups studied. CONCLUSIONS: In hepatitis C, lobular mononuclear cells are mostly macrophages and appear associated with bridging fibrosis. Cirrhotic livers display significantly lower numbers of lobular CD4 and CD8 lymphocytes. This finding could help explain a decrease in immune surveillance and the promotion of neoplastic growth in HCV associated cirrhosis. PMID- 17415342 TI - Management of chronic hepatitis C. AB - Chronic hepatitis C is a major cause of cirrhosis and primary liver cancer (hepatocellular carcinoma). Decompensated cirrhosis or hepatocellular carcinoma secondary to hepatitis C is the first cause of liver transplantation in Europe and in the United States. The prognosis of chronic hepatitis C depends on the progression of fibrosis which determines the risk of developing cirrhosis and its complications. Knowledge of the natural history and the factors associated with the progression of fibrosis is essential for the patient's management. The risk of the progression of fibrosis is difficult to predict in one particular patient. Liver biopsy remains the best test to evaluate the severity of fibrosis, determine its prognosis and discuss the therapeutic options. At present, in a patient with hepatitis C, combined therapy associating pegylated alpha interferon and ribavirin results in a sustained response in approximately 55% of cases. Based on existing results, the sustained virological response with this treatment option appears to be long lasting, to be associated with a histological benefit and is also probably associated with a reduction in the risk of cirrhosis and hepatocellular carcinoma. The management of hepatitis C virus infections must include better knowledge of the natural history of the disease and existing available antiviral treatments (pegylated interferon and ribavirin) as well as in depth knowledge of the aims of treatment, the results obtained, the predictive factors of response and side effects. With close follow-up, doses can be rapidly modified and erythropoietin more frequently administered; new molecules may also be developed in this context. This paper will discuss the natural history, the factors associated with the progression of fibrosis, the predictive factors of response to treatment, and existing and future treatments for hepatitis C. PMID- 17415343 TI - Diagnosis and treatment of chronic hepatitis B: an update. AB - The diagnosis of chronic hepatitis B virus (HBV) infection is made using a combination of serological, virologic, biochemical, and histologic markers. The natural history of HBV infection can be divided into four phases: immune tolerance, immune clearance (HBeAg-positive chronic hepatitis B), inactive HBsAg carrier, and reactivation (HBeAg-negative chronic hepatitis B). Patients in the immune clearance and reactivation phases, with elevated alanine aminotransferase (ALT) and HBV DNA levels, are candidates for antiviral therapy. The primary goal of therapy for chronic hepatitis B is suppression of viral replication, which has been shown to reduce hepatic necroinflammation and retard progression of hepatic fibrosis. Long-term suppression of serum HBV DNA is likely to reduce progression to cirrhosis and hepatic decompensation and decrease the risk of hepatocellular carcinoma. Current antiviral therapy for chronic hepatitis B includes interferon alfa, peginterferon alfa-2a, lamivudine, adefovir, entecavir, and telbivudine. In patients with HBeAg-positive chronic hepatitis B, antiviral treatment is indicated when the serum HBV DNA level is = or > 10(5) copies/mL (20,000 IU/mL) and the ALT level is elevated. For HBeAg-negative patients, the threshold for initiation of therapy is lower, i.e., a serum HBV DNA level = or > 10(4) copies/mL (2,000 IU/mL) in association with an elevated ALT level. The presence of at least moderate necroinflammation and the presence of fibrosis on liver biopsy, which is optional and not mandatory before therapy, may be useful in supporting the decision to initiate therapy, particularly in patients with normal ALT levels. While undergoing therapy, patients require monitoring every 3 to 6 months to ensure compliance and to test for the development of resistance if an oral agent is used. Issues that remain controversial or need to be studied further are the necessity of a baseline liver biopsy, the HBV DNA and ALT thresholds for initiation of therapy, the optimal duration of antiviral therapy, selection of one agent over another, and the role of combination therapy. PMID- 17415344 TI - Evolving concepts in the diagnosis, pathogenesis and treatment of autoimmune hepatitis. AB - The diagnostic criteria for autoimmune hepatitis have been codified, and a scoring system can quantify the strength of the diagnosis. Centrilobular (zone 3) necrosis signifies acute disease, and severe acute and fulminant presentations of autoimmune hepatitis are recognized. The absence of symptoms at presentation may identify some patients who do not require treatment, but therapeutic decisions must be based on disease activity not symptoms, especially since 26-70% of asymptomatic patients become symptomatic. Elderly patients have more advanced disease at presentation, but they respond well to treatment. Antibodies to soluble liver antigen/liver pancreas, asialoglycoprotein receptor, actin, and liver cytosol type 1 have prognostic value. Molecular mimicry between viral and self-antigens is the likely basis for the autoimmune response. Susceptibility alleles optimize antigen presentation. Polymorphisms influence immunocyte activation, counter-regulatory actions within the cytokine milieu, and apoptotic pathways for hepatocyte and immunocyte death. Perturbations in the populations of T regulatory cells and natural killer T cells disrupt immune homeostasis. Cyclosporine, mycophenolate mofetil, and budesonide afford new treatment opportunities, and molecular interventions at critical pathogenic pathways are feasible, especially within the cytokine network. Confident animal models of the human disease and a collaborative network of clinical investigators are the requisites for progress. PMID- 17415345 TI - Diagnostic and therapeutic questions in overlap syndromes of autoimmune hepatitis. AB - The term ''overlap syndromes of liver diseases'' includes coexistence of autoimmune hepatitis (AIH), primary sclerosing cholangitis (PSC), primary biliary cirrhosis (PBC). Due to their unknown etiology, as well as their variable presentation with mixed clinical and biochemical features, these overlap syndromes are often a diagnostic and therapeutic challenge for hepatologists. The most frequent association reported occurs between AIH and PBC. More rare is the overlap between AIH and PSC, typical in young age and often concomitant with an inflammatory bowel disease as ulcerative colitis. The treatment of choice is based on ursodeoxycholic acid and immunosoppressive drugs, used at the same time or consecutively, according to the course of disease. Histological examination seems an important tool, but often does not help for a correct diagnosis due to lack of specificity. Two particular forms of variant syndrome are the so called outlier syndromes, without clear characteristics of overlap: the autoimmune cholangitis, probably a form of PBC anti-mitochondrial antibodies negative, and the hepatitis C virus related with stigmata of autoimmunity, such as nonspecific autoantibodies at low titer. The diagnostic score system elaborated in 1999 by the International AIH Group can help for diagnosis, even if its definite validity is lacking. PMID- 17415346 TI - Alcoholic and nonalcoholic forms of fatty liver disease. AB - Fatty liver (or steatosis) is an increasingly common finding in histolgical evaluation of liver biopsies. It is frequently associated with obesity, diabetes, metabolic syndromes, and/or alcohol abuse. When the steatosis is accompanied by inflammation and fibrosis, the risk of progression to severe liver disease is high. The aim of this paper is to review the clinical features, pathophysiology, natural history, and options for the treatment of nonalcoholic and alcoholic forms of fatty liver disease. PMID- 17415347 TI - Endoscopic frontiers in the field of hepatology. AB - Hepatology is considered a cognitive specialty, but it will not be surprising if a subgroup of future hepatologists (''invasive hepatologists'') performed a variety of advanced endoscopic, laparoscopic, vascular or ablative procedures just like interventional gastroenterologists, interventional radiologists or minimally invasive surgeons. The increase in the prevalence of liver diseases including hepatocellular carcinoma, and effective treatment of end-stage liver disease with liver transplantation has expanded the subspecialty of hepatology into a major specialty. Therefore, it is only natural that some of the trainees in hepatology, familiar with invasive procedures just like their counterparts in gastroenterology, may become subspecialized in invasive aspects of this specialty, traditionally performed by interventional endoscopists, radiologists and surgeons. Moreover, there will be major developments in the management of the complications of liver disease. Endoscopic screening with esophageal capsule endoscopy and, to a lesser extent, ultrathin upper gastrointestinal endoscopy may replace conventional endoscopy. In addition to standard treatments for esophageal varices, removable esophageal stents with expansile pressure may be utilized in refractory variceal hemorrhage. Transjugular intrahepatic portosystemic shunts may be performed by hepatologists. Advances in argon plasma coagulation, cryotherapy and photodynamic therapy may result in novel treatment options for portal hypertensive gastropathy. Single-fiber cholangioscopy will allow for directed endoscopic screening for cholangiocarcinoma and primary sclerosing cholangitis in high-risk individuals. Minilaparoscopy will allow a macroscopic assessment of the liver surface as well as the ability to target specific regions for histopathology, and treatment including radiofrequency ablation of liver cancer. Endoscopic ultrasound (EUS) may provide the potential to directly measure portal vein pressure and this may have a future role in titration and optimization of pharmacological therapy of portal hypertension. EUS and fine needle aspiration may be used for staging hepatocellular and bile duct cancer. Finally, natural orifice transluminal surgery and endoscopic ultrasound-guided angiography may allow for targeted therapies traditionally outside the realm of the hepatologists. PMID- 17415348 TI - Assessment of liver fibrosis in the clinical setting: something is changing? AB - Clinical management of compensated chronic liver diseases (CLD) requires precise definition of the stage of liver fibrosis which is the key histologic predictor of progression to cirrhosis. Several methods are used to assess liver fibrosis. Among those, percutaneous liver biopsy is still the gold standard. However, the recent introduction of liver imaging techniques, the rising of statistical tests able to classify CLD noninvasively, and a reconsideration of its potential complications, have contributed to an audit of the evolving role of liver biopsy. At present, there is an increasing interest for noninvasive approaches to evaluate the stage of liver fibrosis in the clinical work-up of patients with CLD. Transient elastography (FibroScan) is a new, noninvasive method to assess liver stiffness and, consequently, the degree of liver fibrosis. Since its use in the clinical setting is of great interest, further studies should define the exact role of this procedure. PMID- 17415349 TI - Mixed cryoglobulinemia: only hepatitis C virus? PMID- 17415350 TI - Prediction models in urology. PMID- 17415351 TI - Defining the role of overactive bladder treatments in men with lower urinary tract symptoms. PMID- 17415352 TI - Klinefelter syndrome in clinical practice. AB - Klinefelter syndrome is the most common sex-chromosome disorder; it affects approximately one in every 660 men. This syndrome is characterized by the presence of one or more extra X chromosomes, and the karyotype 47,XXY is the most prevalent type. The 'prototypic' man with Klinefelter syndrome has traditionally been described as tall, with narrow shoulders, broad hips, sparse body hair, gynecomastia, small testicles, androgen deficiency, azoospermia and decreased verbal intelligence. A less distinct phenotype has, however, been described. Klinefelter syndrome is an underdiagnosed condition; only 25% of the expected number of patients are diagnosed, and of these only a minority are diagnosed before puberty. Patients with Klinefelter syndrome should be treated with lifelong testosterone supplementation that begins at puberty, to secure proper masculine development of sexual characteristics, muscle bulk and bone structure, and to prevent the long-term deleterious consequences of hypogonadism; however, the optimal testosterone regimen for patients with Klinefelter syndrome remains to be established. PMID- 17415353 TI - The epidemiology of bladder and kidney cancer. AB - Bladder and kidney cancer together account for about 5% of cancers worldwide, and represent the 9th and 14th most common cancers in terms of absolute numbers, respectively. Our knowledge of these two cancers, however, indicates that they have strikingly different etiology. A number of important occupational, lifestyle and genetic factors have been implicated in bladder carcinogenesis, which have greatly increased our understanding of this disease. In some instances, identification of these factors has contributed to the prevention or reduction of exposure to bladder carcinogens. Conversely, the etiology of kidney cancer is less well elucidated, which limits the possibilities for reducing its incidence by this approach. It is likely that much remains to be uncovered about the causes of these two cancers, and these advances will increase our appreciation of the biology of these cancers and perhaps lead to further possibilities for their prevention. In this article, we evaluate the epidemiology of bladder and kidney cancer and also discuss potential priorities for future research. PMID- 17415354 TI - Hereditary urological cancer syndromes. AB - Hereditary urological cancer syndromes are rare, but it is important that they are recognized because they have important prognostic implications; prompt diagnosis can dramatically improve patient outcomes. The urologist or urological oncologist should, therefore, ascertain which tumors of the many seen in clinical practice warrant referral for the opinion of a clinical geneticist. Despite the aggressive natural history of most inherited urological cancer syndromes, organ preserving treatments are desirable because these syndromes predispose affected patients to the formation of multifocal and metachronous tumors. Identification of the molecular mechanisms that underlie carcinogenesis in both familial and sporadic urological cancers has, in some cases, resulted in novel and specifically targeted approaches to therapy. Patients who present with early onset or multiple tumors should be carefully investigated for the presence of a hereditary cancer syndrome, and once a diagnosis is made, appropriate screening should be instigated for family members to enable early detection of tumors both within and outside the urogenital tract. PMID- 17415355 TI - A case of scrotal sarcoidosis that mimicked tuberculosis. AB - BACKGROUND: A 38-year-old man presented with bilateral testicular swelling, night sweats and weight loss with little response to antibiotics. Examination revealed systemic lymphadenopathy and multiple small masses arising from both testes. INVESTIGATIONS: Scrotal ultrasound revealed multiple intratesticular, hypoechoic lesions; chest radiograph and abdominal ultrasound were normal. A CT scan revealed multilevel lymphadenopathies. A Heaf (tuberculin) skin test was negative. Testicular biopsy revealed multiple granulomata, some of which showed patchy central necrosis. DIAGNOSIS: A diagnosis of tuberculosis was initially made. After 3 months of clinical deterioration despite antitubercular drug therapy, however, the diagnosis was changed to sarcoidosis because blood results revealed hypercalcemia, elevated serum angiotensin-converting enzyme, and an elevated erythrocyte sedimentation rate. MANAGEMENT: The patient improved dramatically on corticosteroid therapy, with complete regression of all testicular lesions on imaging after 2 months. Steroids were tapered, then discontinued after 6 months. The patient remained in complete remission, but became oligospermic by the 3 year follow-up. PMID- 17415356 TI - Needles galore, but no place to sew. AB - Large-scale sequencing projects to identify mutations related to cancer and psychiatric disorders will generate a lot of data. But the usefulness of these results will be limited unless we have good models to test their contribution to disease. PMID- 17415357 TI - Changes to US drug safety laws take center stage. PMID- 17415358 TI - Critics slam Russian trial to test pig pancreas for diabetics. PMID- 17415359 TI - Nifty transport device doubles durability of donor organs. PMID- 17415360 TI - Pricey cancer genome project struggles with sample shortage. PMID- 17415361 TI - Controversy over cervical cancer vaccine spurs safety surveillance. PMID- 17415362 TI - Plagiarized report on patent laws shames Indian scientists. PMID- 17415363 TI - Aggressive drug marketing tactics trigger backlash. PMID- 17415365 TI - The sunshine cure. PMID- 17415366 TI - Straight talk from... Robert Klein. PMID- 17415367 TI - NMR-based prediction of cardiovascular risk in diabetes. PMID- 17415369 TI - Netting bacteria in sepsis. PMID- 17415370 TI - Providing AID to p53 mutagenesis. PMID- 17415371 TI - Lung turns to AA (adenosine analogues) to dry out. PMID- 17415372 TI - Immune cells may fend off Alzheimer disease. PMID- 17415373 TI - MicroRNA may have macro effect on sudden death. PMID- 17415374 TI - T effectors outfox T regulators in autoimmunity. PMID- 17415375 TI - Premature poking: impulsivity, cocaine and dopamine. PMID- 17415379 TI - Suitability of a US3-inactivated HSV mutant (L1BR1) as an oncolytic virus for pancreatic cancer therapy. AB - Recently, the use of oncolytic viruses against cancer has attracted considerable attention. We studied the potential of the US3 locus-deficient herpes simplex virus (HSV), L1BR1, for oncolytic virus therapy. Its high specificity and potency indicate that L1BR1 is a promising candidate as a new oncolytic virus against pancreatic cancer. Moreover, the virus exhibited the unique characteristic of increasing apoptosis when used in combination with anticancer drugs. We assessed the feasibility of using the US3 locus-deficient HSV named L1BR1 as a new replication-competent oncolytic virus for the treatment of pancreatic cancer. The US3 locus of HSV has been shown to be a key gene in producing a multifunctional protein kinase that inhibits apoptosis induced by viral infections, chemicals and ultraviolet (UV) light. L1BR1 has been reported to be more than 10 000-fold less virulent than the parental virus in mice. In this study, we examined the tumor specificity and oncolytic effect of this attenuated replication-competent virus, L1BR1, in pancreatic cancers derived from SW1990, Capan2 and Bxpc-3cells compared with the parent virus and other well-known oncolytic herpes viruses (R3616 and hrR3). We also studied the efficacy of L1BR1 for the induction of apoptosis as an attribute of this virus in combination with the anticancer drugs 5FU and cisplatin. The combined treatment of the pancreatic cancer cells with L1BR1 and these anticancer drugs enhanced apoptosis significantly. More importantly, L1BR1 showed the lowest replication capacity in normal human hepatocytes, but the highest tumor-reducing effect in vivo among the oncolytic herpes viruses tested. In addition, L1BR1 significantly increased the induction of apoptosis of cancer cells when treated in combination with anticancer drugs although the parental virus inhibited the induction of apoptosis. These results suggest that L1BR1 is promising as a new anticancer oncolytic virus. PMID- 17415380 TI - Lentiviral vector neutral endopeptidase gene transfer suppresses prostate cancer tumor growth. AB - Neprilysin (neutral endopeptidase, NEP) is a cell surface peptidase whose expression is lost in approximately 50% of prostate cancers (PC). NEP normally functions to inactivate peptides such as bombesin and endothelin-1, and potentiates the effects of the PTEN tumor suppressor via a direct protein-protein interaction. NEP loss contributes to PC progression. We investigated the therapeutic efficacy of using a lentiviral vector system to restore NEP expression in PC cells. Third-generation lentiviral vectors encoding wild-type NEP (L-NEP) or green fluorescent protein (L-GFP) were introduced into NEP deficient 22RV1 PC cells. Cells infected with L-NEP or L-GFP at a multiplicity of infection of 10 demonstrated NEP enzyme activity of 1171.2+/-4.9 and 17.2+/-5.3 pmol/microg/min (P<0.0001), respectively. Cell viability, proliferation and invasion were each significantly inhibited in 22RV1 cells expressing NEP compared with control cells infected with L-GFP (P<0.01). Analysis of known downstream effects of NEP showed NEP-expressing cells exhibiting decreased Akt and focal adhesion kinase phosphorylation and increased PTEN protein expression. Finally, injection of L-NEP into established 22RV1 xenograft tumors significantly inhibited tumor growth (P<0.01). These experiments demonstrate that lentiviral NEP gene transfer is a novel targeted strategy for the treatment of NEP-deficient PC. PMID- 17415381 TI - Transposon-based interferon gamma gene transfer overcomes limitations of episomal plasmid for immunogene therapy of glioblastoma. AB - Despite improvements in gene delivery technology, transient expression of plasmid DNA has limited the efficacy of nonviral vectors applied to cancer gene therapy. We previously developed plasmid DNA vectors capable of transgene integration and long-term expression in human glioblastoma cells by utilizing the Sleeping Beauty (SB) transposable element. In this study, we compared the efficacy of interferon gamma (IFN-gamma) immunogene therapy using episomal or SB vectors in a syngeneic GL261 glioma model. Gene delivery was achieved by intratumoral convection enhanced delivery of DNA/polyethylenimine complexes. Only mice treated with SB transposase-encoding DNA to facilitate chromosomal integration exhibited a significant increase in survival (P<0.05). SB-mediated intratumoral gene transfer caused sustained IFN-gamma expression assessed by reverse transcription polymerase chain reaction, of both vector-derived and endogenous IFN-gamma, whereas expression following episomal plasmid gene transfer was undetectable within 2 weeks. Median survival was enhanced further when SB-mediated IFN-gamma gene transfer was combined with CpG oligodeoxynucleotides as adjuvant therapy. Prolonged survival positively correlated with tumor regression measured by in vivo bioluminescent imaging, and enhanced T-cell activation revealed by the ELISPOT assay. SB appears to improve the efficacy of cytokine gene therapy using nonviral vectors by enhancing the duration of transgene expression. PMID- 17415382 TI - Upregulation and redistribution of integrin alpha6beta4 expression occurs at an early stage in pancreatic adenocarcinoma progression. AB - Pancreatic adenocarcinomas are highly invasive cancers for reasons that are currently unclear. Here we sought to determine if the proinvasive integrin alpha6beta4 may be related to pancreatic adenocarcinoma tumor progression. Expression of integrin alpha6beta4 was analyzed via immunohistochemistry for the beta4 subunit in normal pancreas, pancreatic intraepithelial neoplasia (PanIN) lesions, pancreatic adenocarcinomas and chronic pancreatitis. In normal pancreatic ducts, integrin alpha6beta4 was noted only at the cell's basal interface with the basement membrane. In pancreatic adenocarcinomas, 92% (104/113) demonstrated overexpression of integrin alpha6beta4 and altered localization to the cytoplasm and membranous regions. This pattern of expression was observed in all PanIN lesions as early as PanIN-1A, and was evident in lesions that were juxtapositioned to normal epithelium. In contrast, 93% (13/14) of chronic pancreatitis samples resembled the staining pattern of normal pancreas. When cancer was present in areas of chronic pancreatitis, this altered expression of alpha6beta4 integrin identified the cancer. We conclude that integrin alpha6beta4 is expressed only on the basal surface of ductal cells in normal pancreas and chronic pancreatitis. During pancreatic adenocarcinoma progression, the alpha6beta4 integrin is dramatically overexpressed and displays altered localization at the earliest stages of PanIN, thus representing an early event in pancreatic adenocarcinoma progression. PMID- 17415383 TI - Adaptive beam shaping by controlled thermal lensing in optical elements. AB - We describe an adaptive optical system for use as a tunable focusing element. The system provides adaptive beam shaping via controlled thermal lensing in the optical elements. The system is agile, remotely controllable, touch free, and vacuum compatible; it offers a wide dynamic range, aberration-free focal length tuning, and can provide both positive and negative lensing effects. Focusing is obtained through dynamic heating of an optical element by an external pump beam. The system is especially suitable for use in interferometric gravitational wave interferometers employing high laser power, allowing for in situ control of the laser modal properties and compensation for thermal lensing of the primary laser. Using CO(2) laser heating of fused-silica substrates, we demonstrate a focal length variable from infinity to 4.0 m, with a slope of 0.082 diopter/W of absorbed heat. For on-axis operation, no higher-order modes are introduced by the adaptive optical element. Theoretical modeling of the induced optical path change and predicted thermal lens agrees well with measurement. PMID- 17415384 TI - Double-cavity radiometer for high-flux density solar radiation measurements. AB - A radiometric method has been developed, suitable for both total power and flux density profile measurement of concentrated solar radiation. The high-flux density radiation is collected by a first optical cavity, integrated, and driven to a second optical cavity, where, attenuated, it is measured by a conventional radiometer operating under a stationary irradiation regime. The attenuation factor is regulated by properly selecting the aperture areas in the two cavities. The radiometer has been calibrated by a pulsed solar simulator at concentration levels of hundreds of suns. An optical model and a ray-tracing study have also been developed and validated, by which the potentialities of the radiometer have been largely explored. PMID- 17415385 TI - Analysis of multimask fabrication errors for diffractive optical elements. AB - As design algorithms for diffractive optical elements improve, the limiting factor becomes the fabrication process. It is hoped a better understanding of fabrication errors will allow elements with greater tolerance to be designed. This is important for high-power laser fiber coupling, where hot spots lead to failure. We model seven different fan-out gratings applying misetch, misalignment, and feature rounding. Our main findings are that misetch can lead to improved results, misalignment is strongly asymmetric, and both the pi and pi/2 masks can dominate misalignment. Rounding has a r(2) dependence and potentially can be incorporated into the design stage. Finally we present some experimental data for misalignment. PMID- 17415386 TI - Triple-sensor multiplexed frequency-modulated continuous-wave interferometric fiber-optic displacement sensor. AB - A triple-sensor multiplexed fiber-optic displacement sensor, which can measure the displacements of three different objects or the three-dimensional displacement of a single object, is introduced. The sensor is based on the principles of optical frequency-modulated continuous-wave interference and frequency-division multiplexing. The beat signals from the individual sensors are assigned in the frequency domain and separated with different electrical bandpass filters. The displacements of objects can be determined simultaneously by detecting the phase shifts of the corresponding signals. The cross talk between the individual sensors is evaluated, and an accuracy of 0.08 microm in a dynamic range of 1000 microm is achieved. PMID- 17415387 TI - Analysis of tuning of Bragg wavelength of photowritten fiber Bragg gratings during the inscription process using a biprism. AB - This paper presents an analysis of a simple biprism-based technique that enables inscription of multiple fiber Bragg gratings (FBGs), each with different Bragg wavelength, into a section of an optical fiber. The method offers a tuning range for Bragg wavelength that is comparable to that of the multiple mirror interferometer method. This has an additional advantage over multiple mirror interferometer methods, in that it does not require shifting of the fiber position perpendicular to the fiber axis, over a wide tuning range. Results of an analysis on how such tuning of Bragg wavelengths affects other important parameters of FBGs such as length, bandwidth, and reflectivity have been presented. PMID- 17415388 TI - Accuracy improvement in peak positioning of spectrally distorted fiber Bragg grating sensors by Gaussian curve fitting. AB - To improve measurement accuracy of spectrally distorted fiber Bragg grating temperature sensors, reflection profiles were curve fitted to Gaussian shapes, of which center positions were transformed into temperature information. By applying the Gaussian curve-fitting algorithm in a tunable bandpass filter demodulation scheme, approximately 0.3 degrees C temperature resolution was obtained with a severely distorted grating sensor, which was much better than that obtained using the highest peak search algorithm. A binary search was also used to retrieve the optimal fitting curves with the least amount of processing time. PMID- 17415389 TI - Particle-sizing methods: a stationary-phase-based comparison. AB - Stationary-phase approximation is used to examine and compare the reliability and interpretability of two main methods of particle sizing. The first method, differential light scattering, regards spherical objects having different refractive indices. Theoretical expressions are obtained, enabling the derivation of optical and geometrical properties of the object from its scattering pattern. The second method, automated microscope imaging, is considered with theoretical implications for the finite aperture of the objective lens. It is shown that, besides the well known Rayleigh resolution limit, finite aperture must affect size measurement due to optical properties of the particles. Simulation and experimental results for both differential light scattering and microscope imaging of polystyrene beads are in good agreement with theory. PMID- 17415390 TI - Simplified optical scatterometry for periodic nanoarrays in the near-quasi-static limit. AB - Scatterometry is now proven to be a very powerful technique for measurement of subwavelength periodic structures. However it requires heavy numerical calculations of the scattered optical waves from the structure. For periodic nanoarrays with feature size less than 100 nm, it is possible to simplify this using the Rytov near-quasi-static approximation valid for feature periods only few time less than the wavelength. The validity is investigated by way of comparison with exact numerical results obtained with the eigenfunctions approach. It is shown to be adequate for the determination of the structure parameters from the specularly reflected or transmitted waves and their polarization or ellipsometric properties. The validity of this approach is applied to lamellar nanoscale grating photoresist lines on Si substrate. The high sensitivity of the signals to the structure parameters is demonstrated using wavelengths of only few times the period. PMID- 17415391 TI - Modified Debye model parameters of metals applicable for broadband calculations. AB - The finite-difference time-domain method can provide broadband results if the excitation source is a pulse. This demands that the parameters of modeled materials have to be applicable over broad frequency bands. We optimize the modified Debye model parameters for gold, silver, copper, platinum, and aluminum using a large-scale nonlinear optimization algorithm. The complex relative permittivities calculated using the optimized parameters agree well with experimental values over broad frequency bands. The associated root-mean-square deviations are 0.49%, 3.52%, 4.13%, 1.64%, and 0.66%, respectively. We also provide an example of broadband calculations. The obtained broadband results are verified by a series of steady-state calculations. PMID- 17415392 TI - Attenuation in waveguides on FR-4 boards due to periodic substrate undulations. AB - The guided-mode attenuation associated with optical-interconnect-polymer waveguides fabricated on FR-4 printed-circuit boards is quantified. The rigorous transmission-line network approach is used and the FR-4 substrate is treated as a long-period substrate grating. A quantitative metric for an appropriate matrix truncation is presented. The peaks of attenuation are shown to occur near the Bragg conditions that characterize the leaky-wave stop bands. For a typical 400 microm period FR-4 substrate with an 8 microm corrugation depth, a buffer layer thickness of about 40 microm is found to be needed to make the attenuation negligibly small. PMID- 17415393 TI - Simulation of intensity of a patterned thin-film structure. AB - An enhanced thin-film model is proposed to simulate the reflecting intensity of a patterned thin-film structure received by a CCD. This improved thin-film model can be adopted to simulate the intensity profile of a patterned multilayer and diffractive structure. The model is matched to the image by adding together components from all of the patterned regions in the target and multiplying the sum by a complex reflectance transfer matrix, which represents all of the material characteristics. PMID- 17415394 TI - Scanning near-field optical microscopy signal processing and resolution. AB - To increase the signal-to-noise ratio and to remove the spatially slow varying signals, a lock-in amplifier is often used in scanning probe microscopy. The signal reconstructed from the lock-in data contains the contributions of the evanescent and homogeneous waves that are mixed in the near-field zone (i.e., at a very short distance). The resolution is determined and a method is given to suppress the useless background information. Experimental images of nanoparticles are processed. PMID- 17415395 TI - Pseudorandom phase masks for superresolution imaging from subpixel shifting. AB - We present a method for overcoming the pixel-limited resolution of digital imagers. Our method combines optical point-spread function engineering with subpixel image shifting. We place an optimized pseudorandom phase mask in the aperture stop of a conventional imager and demonstrate the improved performance that can be achieved by combining multiple subpixel shifted images. Simulation results show that the pseudorandom phase-enhanced lens (PRPEL) imager achieves as much as 50% resolution improvement over a conventional multiframe imager. The PRPEL imager also enhances reconstruction root-mean-squared error by as much as 20%. We present experimental results that validate the predicted PRPEL imager performance. PMID- 17415396 TI - Tropospheric ozone differential-absorption lidar using stimulated Raman scattering in carbon dioxide. AB - A UV ozone differential-absorption lidar (DIAL) utilizing a Nd:YAG laser and a single Raman cell filled with carbon dioxide (CO(2)) is designed, developed, and evaluated. The generated wavelengths are 276, 287, and 299 nm, comprising the first to third Stokes lines of the stimulated Raman scattering technique. The correction terms originated from the aerosol extinction, the backscatter, and the absorption by other gases are estimated using a model atmosphere. The experimental results demonstrate that the emitted output energies were 13 mJ/pulse at 276 nm and 287 nm and 5 mJ/pulse at 299 nm, with pump energy of 91 mJ/pulse and a CO(2) pressure of 0.7 MPa. The three Stokes lines account for 44.0% of the available energy. The use of argon or helium as a buffer gas in the Raman cell was also investigated, but this leads to a dramatic decrease in the third Stokes line, which makes this wavelength practically unusable. Our observations confirmed that 30 min of integration were sufficient to observe ozone concentration profiles up to 10 km. Aerosol extinction and backscatter correction are estimated and applied. The aerosol backscatter correction profile using 287 and 299 nm as reference wavelengths is compared with that using 355 nm. The estimated statistical error is less than 5% at 1.5 km and 10% at 2.6 km. Comparisons with the operational carbon-iodine type chemical ozonesondes demonstrate 20% overestimation of the ozone profiles by the DIAL technique. PMID- 17415397 TI - Pump efficiency improvement of a C-band tunable fiber laser using optical circulator and tunable fiber gratings. AB - We propose and demonstrate a tunable fiber laser based on an optical circulator (OC) and two tunable fiber Bragg gratings (TFBGs). The OC acts as a pump power router to improve the pumping efficiency, and a 4% increase in overall conversion efficiency has been observed. The combined tuning spectra range of two TFBGs could cover the entire C-band spectrum from 1530 to 1560 nm. Stable laser output power above 10 dBm is obtained using 1.9 m of erbium-doped fiber and TFBGs with 50% reflectivity. With power equalization by using variable optical attenuators, the power variation is less than 0.1 dB in the whole C band with narrow linewidth of 0.05 nm. A signal-to-noise ratio of 60 dB and a continuous tuning resolution of 0.5 nm have been achieved. The TFBG-based tunable fiber laser can be a promising light source for WDM transmission and fiber sensor applications. PMID- 17415398 TI - Effects of optical feedback in a birefringence-Zeeman dual frequency laser at high optical feedback levels. AB - Optical feedback effects are studied in a birefringence-Zeeman dual frequency laser at high optical feedback levels. The intensity modulation features of the two orthogonally polarized lights are investigated in both isotropic optical feedback (IOF) and polarized optical feedback (POF). In IOF, the intensities of both beams are modulated simultaneously, and four zones, i.e., the e-light zone, the o-light and e-light zone, the o-light zone, and the no-light zone, are formed in a period corresponding to a half laser wavelength displacement of the feedback mirror. In POF, the two orthogonally polarized lights will oscillate alternately. Strong mode competition can be observed, and it affects the phase difference between the two beams greatly. The theoretical analysis is presented, which is in good agreement with the experimental results. The potential use of the experimental results is also discussed. PMID- 17415399 TI - Passively Q-switched Yb3+:YCa4O(BO3)3 laser with InGaAs quantum wells as saturable absorbers. AB - A diode-pumped Yb:YCOB laser at 1086 nm is passively Q switched by using InGaAs quantum wells as saturable absorbers and utilizing the Bragg mirror structure as an output coupler. With an absorbed pump power of 9.2 W the laser produces pulses of 100 ms duration with average pulse energy of as much as 165 microJ at a pulse repetition rate of 7 kHz. PMID- 17415400 TI - Model-based far-field alignment algorithm for Gaussian beamlike single-mode optical devices. AB - Single-mode device-to-fiber alignment automation is usually achieved with a classical mathematical optimization approach. We present a different approach, which is based on the identification of particular intrinsic characteristics of the coupled optical power and on estimating residual axial, transverse, and angular misalignments in the far field. Such a model-based approach is based on the physical nature of the optical coupling phenomenon and can replace or be complementary to already known automated alignment methods. An alignment algorithm is described and validated experimentally using two single-mode fibers as Gaussian beam emitter and receiver. PMID- 17415401 TI - Phase determination for a Fourier transform infrared spectrometer in emission mode. AB - Beam-splitter emission strongly influences the spectra measured with a Fourier transform spectrometer (FTS) as it affects the entire phase behavior, in particular in emission spectroscopy. The various radiation contributions of the scene and the FTS itself have different phases in the complex spectrum. As a specific feature, the radiation of the beam splitter is rotated by approximately pi/2 relative to the scene effective radiation. By classical methods of phase correction, the radiation components of different phases are mixed in the complex plane, which may lead to serious errors in the calibrated spectra. For this reason, the nature of the FTS phase has been studied, and a statistical phase determination method has been developed. It allows us to determine the phase function of the scene by minimizing the correlation between the imaginary and the real parts of the complex spectrum and by reducing the variance of the imaginary part. Thus phase accuracies of 10 to 30 mrad can be achieved. In addition, the remaining error of the phase can be calculated for each individual spectrum. The total phase error and its effect on the spectra are discussed. PMID- 17415402 TI - Improvement of amplified spontaneous emission performance by doping tris(8 hydroxyquinoline) aluminum (Alq3) in dye-doped polymer thin films. AB - A well-known red fluorescent dye 4-(dicy-anomethylene)-2-t-butyl-6(1,1,7,7 tetramethyljulolidyl-9-enyl)-4H-pyran (DCJTB) was codoped with an electron transport organic molecule tris(8-hydroxyquinoline) aluminum (Alq(3)) in a host matrix of polystyrene (PS), and the amplified spontaneous emission (ASE) was studied by optically pumping. It was found that the ASE performance was significantly improved by the introduction of Alq(3). The Alq(3):DCJTB:PS blending thin films showed a low threshold (2.4 microJ/pulse) and a high net gain coefficient (109.95 cm(-1)) compared with the pure DCJTB:PS system (threshold of 15.2 microJ/pulse and gain of 35.94 cm(-1)). The improvement of the ASE performance was considered to be attributable to the effective Foster energy transfer from Alq(3) to DCJTB. Our results demonstrate that the Alq(3):DCJTB could be a promising candidate as gain medium for red organic diode lasers. PMID- 17415403 TI - Two-dimensional biosensor arrays based on surface plasmon resonance phase imaging. AB - We present a biosensor design based on capturing the two-dimensional (2D) phase image of surface plasmon resonance (SPR). This 2D SPR imaging technique may enable parallel label-free detection of multiple analytes and is compatible with the microarray chip platform. This system uses our previously reported differential phase measurement approach, in which 2D phase maps obtained from the signal (P) and reference (S) polarizations are compared pixel by pixel. This technique greatly improves detection resolution as the subtraction step can eliminate measurement fluctuations caused by external disturbances as they essentially appear in both channels. Unlike conventional angular SPR systems, in which illumination from a range of angles must be used, phase measurement requires illumination from only one angle, thus making it well suited for 2D measurement. Also, phase-stepping introduced from a moving mirror provides the necessary modulation for accurate detection of the phase. In light of the rapidly increasing need for fast real-time detection, quantification, and identification of a range of proteins for various biomedical applications, our 2D SPR phase imaging technique should hold a promising future in the medical device market. PMID- 17415404 TI - Monte Carlo and discrete-ordinate simulations of spectral radiances in a coupled air-tissue system. AB - We perform a detailed comparison study of Monte Carlo (MC) simulations and discrete-ordinate radiative-transfer (DISORT) calculations of spectral radiances in a 1D coupled air-tissue (CAT) system consisting of horizontal plane-parallel layers. The MC and DISORT models have the same physical basis, including coupling between the air and the tissue, and we use the same air and tissue input parameters for both codes. We find excellent agreement between radiances obtained with the two codes, both above and in the tissue. Our tests cover typical optical properties of skin tissue at the 280, 540, and 650 nm wavelengths. The normalized volume scattering function for internal structures in the skin is represented by the one-parameter Henyey-Greenstein function for large particles and the Rayleigh scattering function for small particles. The CAT-DISORT code is found to be approximately 1000 times faster than the CAT-MC code. We also show that the spectral radiance field is strongly dependent on the inherent optical properties of the skin tissue. PMID- 17415405 TI - Combined enhanced fluorescence and label-free biomolecular detection with a photonic crystal surface. AB - A 2D photonic crystal surface with a different period in each lateral direction is demonstrated to detect biomolecules using two distinct sensing modalities. The sensing mechanisms both rely on the generation of a resonant reflection peak at one of two specific wavelengths, depending on the polarization of light that is incident on the photonic crystal. One polarization results in a resonant reflection peak in the visible spectrum to coincide with the excitation wavelength of a fluorophore, while the orthogonal polarization results in a resonant reflection peak at an infrared wavelength which is used for label-free detection of adsorbed biomolecules. The photonic crystal resonance for fluorescence excitation causes enhanced near fields at the structure surface, resulting in increased signal from fluorophores within 100 nm of the device surface. Label-free detection is performed by illuminating the photonic crystal with white light and monitoring shifts in the peak reflected wavelength of the infrared resonance with a high-resolution imaging detection instrument. Rigorous coupled-wave analysis was used to determine optimal dimensions for the photonic crystal structure, and devices were fabricated using a polymer-based nanoreplica molding approach. Fluorescence-based and label-free detection were demonstrated using arrays of spots of dye-conjugated streptavidin. Quantification of the fluorescent signal showed that the fluorescence output from protein spots on the photonic crystal was increased by up to a factor of 35, and deposited spots were also imaged in the label-free detection mode. PMID- 17415406 TI - Metallic film optimization in a surface plasmon resonance biosensor by the extended Rouard method. AB - The extended Rouard method is applied to the computation of a multi-absorbing layer system for the optimization of surface plasmon resonance (SPR) sensors. Specifically, the effect of the properties of a metallic layer on the shape of the reflectivity and sensitivity curve is demonstrated in the case of a Kretschmann configuration. This theoretical investigation allows us to establish the best optical properties of the metal to obtain a localized SPR, given the illuminating beam properties. Toward the development of a sensitive biosensor based on SPR, we quantify the changes in reflectivity of such an optical biosensor induced by the deposition of a nanometric biochemical film as a function of the metal film characteristics and the illumination operating conditions. The sensitivity of the system emphasizes the potential of such biophotonic technology using metallic multilayer configurations, especially with envisioned metamaterials. PMID- 17415407 TI - Deductive biocomputing. AB - BACKGROUND: As biologists increasingly rely upon computational tools, it is imperative that they be able to appropriately apply these tools and clearly understand the methods the tools employ. Such tools must have access to all the relevant data and knowledge and, in some sense, "understand" biology so that they can serve biologists' goals appropriately and "explain" in biological terms how results are computed. METHODOLOGY/PRINCIPAL FINDINGS: We describe a deduction based approach to biocomputation that semiautomatically combines knowledge, software, and data to satisfy goals expressed in a high-level biological language. The approach is implemented in an open source web-based biocomputing platform called BioDeducta, which combines SRI's SNARK theorem prover with the BioBike interactive integrated knowledge base. The biologist/user expresses a high-level conjecture, representing a biocomputational goal query, without indicating how this goal is to be achieved. A subject domain theory, represented in SNARK's logical language, transforms the terms in the conjecture into capabilities of the available resources and the background knowledge necessary to link them together. If the subject domain theory enables SNARK to prove the conjecture--that is, to find paths between the goal and BioBike resources--then the resulting proofs represent solutions to the conjecture/query. Such proofs provide provenance for each result, indicating in detail how they were computed. We demonstrate BioDeducta by showing how it can approximately replicate a previously published analysis of genes involved in the adaptation of cyanobacteria to different light niches. CONCLUSIONS/SIGNIFICANCE: Through the use of automated deduction guided by a biological subject domain theory, this work is a step towards enabling biologists to conveniently and efficiently marshal integrated knowledge, data, and computational tools toward resolving complex biological queries. PMID- 17415408 TI - Phase 1 study of two merozoite surface protein 1 (MSP1(42)) vaccines for Plasmodium falciparum malaria. AB - OBJECTIVES: To assess the safety and immunogenicity of two vaccines, MSP1(42) FVO/Alhydrogel and MSP1(42)-3D7/Alhydrogel, targeting blood-stage Plasmodium falciparum parasites. DESIGN: A Phase 1 open-label, dose-escalating study. SETTING: Quintiles Phase 1 Services, Lenexa, Kansas between July 2004 and November 2005. PARTICIPANTS: Sixty healthy malaria-naive volunteers 18-48 y of age. INTERVENTIONS: The C-terminal 42-kDa region of merozoite surface protein 1 (MSP1(42)) corresponding to the two allelic forms present in FVO and 3D7 P. falciparum lines were expressed in Escherichia coli, refolded, purified, and formulated on Alhydrogel (aluminum hydroxide). For each vaccine, volunteers in each of three dose cohorts (5, 20, and 80 microg) were vaccinated at 0, 28, and 180 d. Volunteers were followed for 1 y. OUTCOME MEASURES: The safety of MSP1(42) FVO/Alhydrogel and MSP1(42)-3D7/Alhydrogel was assessed. The antibody response to each vaccine was measured by reactivity to homologous and heterologous MSP1(42), MSP1(19), and MSP1(33) recombinant proteins and recognition of FVO and 3D7 parasites. RESULTS: Anti-MSP1(42) antibodies were detected by ELISA in 20/27 (74%) and 22/27 (81%) volunteers receiving three vaccinations of MSP1(42) FVO/Alhydrogel or MSP1(42)-3D7/Alhydrogel, respectively. Regardless of the vaccine, the antibodies were cross-reactive to both MSP1(42)-FVO and MSP1(42)-3D7 proteins. The majority of the antibody response targeted the C-terminal 19-kDa domain of MSP1(42), although low-level antibodies to the N-terminal 33-kDa domain of MSP1(42) were also detected. Immunofluorescence microscopy of sera from the volunteers demonstrated reactivity with both FVO and 3D7 P. falciparum schizonts and free merozoites. Minimal in vitro growth inhibition of FVO or 3D7 parasites by purified IgG from the sera of the vaccinees was observed. CONCLUSIONS: The MSP1(42)/Alhydrogel vaccines were safe and well tolerated but not sufficiently immunogenic to generate a biologic effect in vitro. Addition of immunostimulants to the Alhydrogel formulation to elicit higher vaccine-induced responses in humans may be required for an effective vaccine. PMID- 17415409 TI - Mechanosensation and Transduction in Osteocytes. AB - There may be no single mechanoreceptor in osteocytes, but instead a combination of events that has to be triggered for mechanosensation and transduction of signal to occur. Possibilities include shear stress along dendritic processes and/or the cell body, cell deformation in response to strain, and primary cilia. These events could occur independently or simultaneously to activate mechanotransduction. Signal initiators include calcium channel activation and ATP, nitric oxide, and prostaglandin release. Means of signal transfer include gap junctions and hemichannels, and the release of signaling molecules into the bone fluid. Questions remain regarding the magnitude of strain necessary to induce an osteocyte response, how the response propagates within the osteocyte network, and the timing involved in the initiation of bone resorption and/or formation on the bone surface. Mechanical loading in the form of shear stress is clearly involved not only in mechanosensation and transduction, but also in osteocyte viability. It remains to be determined if mechanical loading can also affect mineral homeostasis and mineralization, which are newly recognized functions of osteocytes. PMID- 17415410 TI - Sulfide-Arrested Growth of Gold Nanorods. AB - The growth of gold nanorods can be arrested at intermediate stages by treatment with Na(2)S, providing greater control over their optical resonances. Nanorods prepared by the seeded reduction of AuCl(4) in aqueous cetyltrimethylammonium bromide solutions in the presence of AgNO(3) typically exhibit a gradual blueshift in longitudinal plasmon resonance, over a period of hours to days. This "optical drift" can be greatly reduced by adding millimolar concentrations of Na(2)S to quench nanorod growth, with an optimized sulfur:metal ratio of 4:1. The sulfide-treated nanorods also experience a marked redshift as a function of Na(2)S concentration to produce stable plasmon resonances well into the near infrared. Sulfide treatment permitted a time-resolved analysis of nanorod growth by transmission electron microscopy, revealing two distinct periods: an initial growth burst (t < 15 min) that generates dumbbell-shaped nanorods with flared ends and a slower phase (t > 30 min) favoring growth around the midsection, leading to nanorods with the more familiar oblate geometry. The blueshift in plasmon resonance that accompanies the dumbbell-to-oblate shape transition correlates more strongly with changes in the length-to-midsection (L/D(1)) ratio rather than the length-to-end width (L/D(2)) ratio, based on the empirical relationship introduced by El-Sayed and co-workers. PMID- 17415411 TI - Relationships between probabilistic Boolean networks and dynamic Bayesian networks as models of gene regulatory networks. AB - A significant amount of attention has recently been focused on modeling of gene regulatory networks. Two frequently used large-scale modeling frameworks are Bayesian networks (BNs) and Boolean networks, the latter one being a special case of its recent stochastic extension, probabilistic Boolean networks (PBNs). PBN is a promising model class that generalizes the standard rule-based interactions of Boolean networks into the stochastic setting. Dynamic Bayesian networks (DBNs) is a general and versatile model class that is able to represent complex temporal stochastic processes and has also been proposed as a model for gene regulatory systems. In this paper, we concentrate on these two model classes and demonstrate that PBNs and a certain subclass of DBNs can represent the same joint probability distribution over their common variables. The major benefit of introducing the relationships between the models is that it opens up the possibility of applying the standard tools of DBNs to PBNs and vice versa. Hence, the standard learning tools of DBNs can be applied in the context of PBNs, and the inference methods give a natural way of handling the missing values in PBNs which are often present in gene expression measurements. Conversely, the tools for controlling the stationary behavior of the networks, tools for projecting networks onto sub networks, and efficient learning schemes can be used for DBNs. In other words, the introduced relationships between the models extend the collection of analysis tools for both model classes. PMID- 17415412 TI - Ewing sarcoma gene EWS is essential for meiosis and B lymphocyte development. AB - Ewing sarcoma gene EWS encodes a putative RNA-binding protein with proposed roles in transcription and splicing, but its physiological role in vivo remains undefined. Here, we have generated Ews-deficient mice and demonstrated that EWS is required for the completion of B cell development and meiosis. Analysis of Ews(-/-) lymphocytes revealed a cell-autonomous defect in precursor B lymphocyte (pre-B lymphocyte) development. During meiosis, Ews-null spermatocytes were deficient in XY bivalent formation and showed reduced meiotic recombination, resulting in massive apoptosis and complete arrest in gamete maturation. Inactivation of Ews in mouse embryonic fibroblasts resulted in premature cellular senescence, and the mutant animals showed hypersensitivity to ionizing radiation. Finally, we showed that EWS interacts with lamin A/C and that loss of EWS results in a reduced lamin A/C expression. Our findings reveal essential functions for EWS in pre-B cell development and meiosis, with proposed roles in DNA pairing and recombination/repair mechanisms. Furthermore, we demonstrate a novel role of EWS in cellular senescence, possibly through its interaction and modulation of lamin A/C. PMID- 17415413 TI - Inhibition of TGF-beta with neutralizing antibodies prevents radiation-induced acceleration of metastatic cancer progression. AB - We investigated whether TGF-beta induced by anticancer therapies accelerates tumor progression. Using the MMTV/PyVmT transgenic model of metastatic breast cancer, we show that administration of ionizing radiation or doxorubicin caused increased circulating levels of TGF-beta1 as well as increased circulating tumor cells and lung metastases. These effects were abrogated by administration of a neutralizing pan-TGF-beta antibody. Circulating polyomavirus middle T antigen expressing tumor cells did not grow ex vivo in the presence of the TGF-beta antibody, suggesting autocrine TGF-beta is a survival signal in these cells. Radiation failed to enhance lung metastases in mice bearing tumors that lack the type II TGF-beta receptor, suggesting that the increase in metastases was due, at least in part, to a direct effect of TGF-beta on the cancer cells. These data implicate TGF-beta induced by anticancer therapy as a pro-metastatic signal in tumor cells and provide a rationale for the simultaneous use of these therapies in combination with TGF-beta inhibitors. PMID- 17415414 TI - Mice lacking inhibitory leptin receptor signals are lean with normal endocrine function. AB - The adipose-derived hormone, leptin, acts via its receptor (LRb) to convey the status of body energy stores to the brain, decreasing feeding and potentiating neuroendocrine energy expenditure. The failure of high levels of leptin in most obese individuals to promote weight loss defines a state of diminished responsiveness to increased leptin, termed leptin resistance. Leptin stimulates the phosphorylation of several tyrosine residues on LRb to mediate leptin action. We homologously replaced LRb in mice with a receptor with a mutation in one of these sites (Tyr985) in order to examine its role in leptin action and signal attenuation in vivo. Mice homozygous for this mutation are neuroendocrinologically normal, but females demonstrate decreased feeding, decreased expression of orexigenic neuropeptides, protection from high-fat diet induced obesity, and increased leptin sensitivity in a sex-biased manner. Thus, leptin activates autoinhibitory signals via LRb Tyr985 to attenuate the anti adiposity effects of leptin, especially in females, potentially contributing to leptin insensitivity in obesity. PMID- 17415415 TI - Crosstalk between Gi and Gq/Gs pathways in airway smooth muscle regulates bronchial contractility and relaxation. AB - Receptor-mediated airway smooth muscle (ASM) contraction via G(alphaq), and relaxation via G(alphas), underlie the bronchospastic features of asthma and its treatment. Asthma models show increased ASM G(alphai) expression, considered the basis for the proasthmatic phenotypes of enhanced bronchial hyperreactivity to contraction mediated by M(3)-muscarinic receptors and diminished relaxation mediated by beta(2)-adrenergic receptors (beta(2)ARs). A causal effect between G(i) expression and phenotype has not been established, nor have mechanisms whereby G(i) modulates G(q)/G(s) signaling. To delineate isolated effects of altered G(i), transgenic mice were generated overexpressing G(alphai2) or a G(alphai2) peptide inhibitor in ASM. Unexpectedly, G(alphai2) overexpression decreased contractility to methacholine, while G(alphai2) inhibition enhanced contraction. These opposite phenotypes resulted from different crosstalk loci within the G(q) signaling network: decreased phospholipase C and increased PKCalpha, respectively. G(alphai2) overexpression decreased beta(2)AR-mediated airway relaxation, while G(alphai2) inhibition increased this response, consistent with physiologically relevant coupling of this receptor to both G(s) and G(i). IL-13 transgenic mice (a model of asthma), which developed increased ASM G(alphai), displayed marked increases in airway hyperresponsiveness when G(alphai) function was inhibited. Increased G(alphai) in asthma is therefore a double-edged sword: a compensatory event mitigating against bronchial hyperreactivity, but a mechanism that evokes beta-agonist resistance. By selective intervention within these multipronged signaling modules, advantageous G(s)/G(q) activities could provide new asthma therapies. PMID- 17415416 TI - Modulation of adverse cardiac remodeling by STARS, a mediator of MEF2 signaling and SRF activity. AB - Cytoskeletal proteins have been implicated in the pathogenesis of cardiomyopathy, but how the cytoskeleton influences the transcriptional alterations associated with adverse cardiac remodeling remains unclear. Striated muscle activator of Rho signaling (STARS) is a muscle-specific actin-binding protein localized to the Z disc that activates serum response factor-dependent (SRF-dependent) transcription by inducing nuclear translocation of the myocardin-related SRF coactivators MRTF A and -B. We show that STARS expression is upregulated in mouse models of cardiac hypertrophy and in failing human hearts. A conserved region of the STARS promoter containing an essential binding site for myocyte enhancer factor-2 (MEF2), a stress-responsive transcriptional activator, mediates cardiac expression of STARS, which in turn activates SRF target genes. Forced overexpression of STARS in the heart sensitizes the heart to pressure overload and calcineurin signaling, resulting in exaggerated deterioration in cardiac function in response to these hypertrophic stimuli. These findings suggest that STARS modulates the responsiveness of the heart to stress signaling by functioning as a cytoskeletal intermediary between MEF2 and SRF. PMID- 17415417 TI - Interaction of acetylcholinesterase inhibitor donepezil with ionic channels of the neuronal membrane. AB - The effects of donepezil on voltage-dependent Ca(2+)- and low-threshold K(+) current were studied on isolated molluscan neurons using two-electrode voltage clamp technique. Donepezil reduced the amplitude of voltage-dependent Ca(2+) current (IC(50)=7.9 microM) and shifted the current-voltage relationships toward hyperpolarization. Donepezil in low concentration (5 microM) increased, while in higher concentrations (>or=10 microM) decreased the low-threshold K(+)-current. PMID- 17415418 TI - Intralysosomal accumulation of gadolinium and lysosomal damage during selective depression of liver macrophages in vivo. AB - Kinetics of gadolinium accumulation was studied by inductively coupled plasma emission spectroscopy after intravenous injection of this agent (7.5 mg/kg) to CBA mice. Gadolinium exhibits lysosomotropic properties (long-term selective accumulation in lysosomes in vivo). Gadolinium uptake by hepatic cells attained maximum 1 h after its intravenous injection and remained at this level during the next day. Accumulation of gadolinium in hepatocytic lysosomes disturbed their osmotic properties (as was seen from the increase in free acid phosphatase activity, which persisted for 19 days). Serum activities of beta-D-galactosidase and beta-D-glucuronidase also increased (24-72 h and day 19). Selective depression of liver macrophages (24-48 h) was accompanied by a decrease in serum chitotriosidase activity. We conclude that accumulation of gadolinium in lysosomes of liver macrophages leads to their damage and elimination of a certain population of macrophages (primarily large cells). Changes in activity of serum lysosomal enzymes also reflect repopulation of liver macrophages. PMID- 17415419 TI - Effect of routine therapy and selective plasmapheresis on electrical properties of cryoglobulins during ischemic stroke. AB - The charge properties of cryoglobulins were examined during the first 21 days of ischemic stroke (atherothrombotic and cardioembolic). Routine drug therapy produced no effect on the charge of cryoglobulins. Plasmapheresis significantly modified the electrokinetic parameters of cryoglobulins during atherothrombotic stroke and produced less pronounced effect on these proteins during cardioembolic stroke. PMID- 17415420 TI - Peculiarities of autonomic regulation assessed by variability of hemodynamic parameters in rats with different stress resistance. AB - Analysis of contribution of sympathetic and parasympathetic systems into heart rate variability carried out using atenolol and atropine showed that August rats are characterized by enhanced tone of the sympathetic system and reduced tone of the parasympathetic system compared to Wistar rats. Reduced tone of the parasympathetic system is also confirmed by lower sensitivity of the baroreflex. Blockade of NO synthesis with Nw-nitro-L-arginine more markedly increased blood pressure variability in August rats compared to Wistar rats. The data attest to a certain rigidity of the autonomic cardiovascular regulation in August rats. PMID- 17415421 TI - Effect of activated protein C on secretory activity of rat peritoneal mast cells. AB - Generation of thrombin and activated protein C in the inflammatory focus was demonstrated in rats with experimental acute peritonitis. The contents of thrombin and activated protein C peaked by the 30th and 120th minute of inflammation, respectively. In vitro study showed a decrease in spontaneous and compound 48/80-induced secretion of beta-hexosaminidase by peritoneal mast cells under the influence of activated protein C in low concentrations. The antiinflammatory effect of protein C in the focus of acute peritonitis is probably realized through NO release from peritoneal mast cells. This conclusion is derived from the data that L-NAME abolishes the protective effect of activated protein C. PMID- 17415422 TI - Peculiarities of active avoidance conditioning in rats with various forms of inherited arterial hypertension. AB - Peculiarities of active avoidance conditioning were studied in NISAG rats (hereditary stress-induced arterial hypertension) and spontaneously hypertensive SHR rats. Conditioning was successful in 90% normotensive Wistar rats and in only 9.1% NISAG rats. Hypertensive SHR rats were intermediate between Wistar and NISAG rats by their learning capacity (66.7%). Our results suggest that differences in learning capacity of hypertensive rats are determined by genetic characteristics of animal behavior and emotional state, rather than blood pressure elevation. PMID- 17415423 TI - Effect of rapid slight cooling of the skin in various phases of immunogenesis on the immune response. AB - Slight cooling had no effect on heat emission and heat production, but modulated the immune response to antigen in animals. Changes in the immune response upon rapid slight cooling of the skin (by 1.5 degrees C) depended on the phase of immunogenesis corresponding to cold exposure. When cooling was performed immediately after immunization, antibody production increased in the spleen and blood, while antigen binding in the spleen remained unchanged. Cold exposure on day 5 after antigen treatment as well as immunization at the peak of cooling did not modulate antibody production, but increased antigen binding in the spleen. Our findings attest to an important role of the temperature factor in the formation of the immune response, which should be taken into account during vaccination. PMID- 17415424 TI - Effect of endotoxemia on skin antioxidant enzymes under experimental conditions. AB - Intraperitoneal injection of bacterial lipopolysaccharide in a dose of 1 mg/kg was followed by prestimulation of whole blood leukocytes in rats. Activities of peroxide- and lipoperoxide-utilizing antioxidant enzymes glutathione peroxidase, glutathione S-transferase, and catalase increased 1 day after lipopolysaccharide administration, while the content of malonic dialdehyde in the skin remained unchanged. PMID- 17415425 TI - Ceruloplasmin prevents hemostatic disorders during experimental hyperammonemia. AB - Experimental hyperammonemia in rats was accompanied by hemostatic disorders manifesting in coagulopathy (activation of the intrinsic pathway of blood coagulation) and suppression of platelet function. Ceruloplasmin in a total dose of 60 mg/kg effectively normalized coagulation hemostasis and functional activity of platelets by improving secretory processes in platelets and increasing aggregation rate. PMID- 17415426 TI - Effect of acoustic stimulation on cell composition of auditory brain structures in Krushinskii-Molodkina rats. AB - We studied the effect of acoustic stimulation on cell composition of auditory brain structures in Krushinskii-Molodkina rats. Significant structural changes in the inferior colliculi of rats with high seizure activity were revealed 1 month after acoustic stimulation. Therefore, this brain structure plays a role in the development of audiogenic epileptic activity. PMID- 17415427 TI - Effect of mexidol and nitroglycerine on iron-sulfur centers, cytochrome P-450, and nitric oxide formation in liver tissue of experimental animals. AB - In experiments on mice we studied the effect of individual or combined treatment with mexidol and nitroglycerine on iron-sulfur centers of the mitochondrial respiratory chain, cytochrome P-450 of the endoplasmic reticulum, and nitric oxide formation in the liver tissue. Mexidol had a potent effect on these parameters and protected iron-sulfur centers from oxidation, including that induced by nitroglycerine. PMID- 17415428 TI - Antioxidant and neuroprotective properties of N-docosahexaenoyl dopamine. AB - N-Docosahexaenoyl dopamine exhibited antioxidant activity in the test with a stable oxygen radical galvinoxyl. This compound produced a dose-dependent protective effect on cultured granular cells from rat cerebellum under conditions of oxidative stress. N-Docosahexaenoyl dopamine decelerated the development of symptoms of Parkinson's disease in mice receiving neurotoxin 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine. PMID- 17415429 TI - Stimulatory effect of fragments from transcribed region of ribosomal repeat on human peripheral blood lymphocytes. AB - Fragments from the transcribed region of the ribosomal repeat include considerable amounts of unmethylated CpG DNA motifs. These motifs activate immune cells via the interaction with Toll receptors. In vitro experiments confirmed the stimulatory effect of transcribed region of ribosomal repeat on human lymphocytes. Culturing of lymphocytes in a medium containing 2-20,000 ng/ml fragments from transcribed region of ribosomal repeat was accompanied by structural changes in the nucleus in a considerable number of cells. These changes manifested in translocation of pericentromeric heterochromatin from the membrane to the center of the nucleus and activation of the nucleolus and were accompanied by a significant increase in interleukin-6 production and slight stimulation of tumor necrosis factor-a synthesis. The transcribed region of the ribosomal repeat and E. coli DNA had various effects on quantitative parameters of lymphocytes. Our results suggest the existence of mechanisms of stimulation not mediated by the interaction of CpG DNA motifs with Toll receptors. PMID- 17415430 TI - Autofluorescence of low-density lipoproteins modified as a result of autooxidation. AB - Autooxidation of low-density lipoproteins during incubation at 37 degrees C was accompanied by accumulation of LPO products, decrease in UV autofluorescence (FUV), and increase in autofluorescence in the visible band (FVIS). The degree of low-density lipoprotein modification was estimated by calculating the FVIS/FUV ratio. A positive correlation was revealed between this ratio and concentration of thiobarbituric acid-reactive LPO products (r=0.76, p<0.001). Autooxidation of low-density lipoproteins increased availability of tryptophanyls for fluorescence quenchers and inductive resonance energy transfer from tryptophanyls to adducts formed in the reaction of apoprotein and LPO products. These changes probably play a role in the decrease in FUV. PMID- 17415431 TI - Melphalan in ultralow doses decreases the severity of experimental colitis in mice. AB - We studied the effect of melphalan in ultralow doses on mice with experimental colitis induced by substitution of drinking water for 5% dextran sulfate. Daily treatment with melphalan in a dose of 0.025 mg/kg improved the general state of animals. The influence of melphalan was evaluated by quantitative clinical, pathomorphological, and laboratory parameters. Melphalan had a local and systemic antiinflammatory effect. PMID- 17415432 TI - Protective effect of energy metabolism regulators in alteration of gravitation load under experimental conditions. AB - Mitochondrial substrate-based preparations corrected disorders, caused by long term exposure to abnormal gravitation vector in head-down tilt (hanging) test in rats. The preparations produced systemic and polyorgan protective effects consisting in correction of the blood prooxidant/antioxidant balance, energy metabolism in musculus soleus, and minimization of morphological changes in the liver and kidneys. PMID- 17415433 TI - Antinecrotic and antioxidant effects of superoxide dismutase during skin ischemia. AB - Antinecrotic activity of SOD was studied in rats with experimental skin ischemia. Treatment with SOD increased activity of endogenous SOD in skin homogenates (by 70 and 26% compared to the ischemic and intact skin, respectively). However, the rate of superoxide anion generation remained unchanged after SOD treatment. Creatine phosphate content and NAD/NADH redox potential increased by 16 and 21%, respectively, on day 3 after SOD administration. The increase in functional activity of the energy supply system and rise in the reserve capacity of the antioxidant protection system contribute to inhibition of lactate dehydrogenase and creatine phosphokinase and decrease in the cytolysis index under the influence of SOD. Our results indicate that SOD produces the antinecrotic effect and holds much promise for the therapy of skin ischemia. PMID- 17415434 TI - Negative chronotropic effect of cannabinoids and their water-soluble emulsion is related to activation of cardiac CB1 receptors. AB - Intravenous injection of cannabinoids dissolved in cremophore EL:ethanol:NaCl mixture and water-soluble emulsion of the same cannabinoids caused identical negative chronotropic effects in chloralose-narcotized rats. Selective CB1 and CB2 receptor antagonist HU-210 also induced a negative chronotropic effect in rats, while pre-injection of CB1 receptor antagonist SR 141716A completely abolished this effect of HU-210. Selective CB2 receptor antagonist SR 144528 had no effect on HU-210-induced bradycardia. Preinjection of ganglioblocker hexamethonium also did not abolish the negative chronotropic effect of HU-210 and ACPA. Perfusion of isolated rat heart with Krebs-Henseleit solution containing HU 210 in a final concentration of 100 nM reduced heart rate. It was shown that the negative chronotropic effect of cannabinoids is mediated through activation of cardiac CB1 receptors. PMID- 17415435 TI - Gastroprotective effect of natural non-starch polysaccharides. AB - Experiments on mice and rats with modeled neurogenic and indomethacin-induced injuries to the gastric mucosa showed that preventive courses of calcium alginate, calcium pectate, and low-esterified pectin improved the resistance of the gastroduodenal mucosa to the destructive effect of the above factors. PMID- 17415436 TI - Effects of L-, D-, and DL-carnitine on morphometric parameters of skeletal muscle and exercise performance of laboratory animals receiving carnitine-deficient diet. AB - Serum concentration of L-carnitine, the mean thickness of the skeletal muscle fiber, and exercise performance in the forced swimming test decreased in rats receiving a carnitine-deficient diet. Treatment with L-carnitine compensated for carnitine deficiency, while racemate and D-stereoisomer did not increase its level. L-Carnitine, but not racemate and D-stereoisomer, promoted recovery of the skeletal muscle fiber thickness and exercise performance of rats. PMID- 17415437 TI - Analysis of antioxidant properties of chitosan and its oligomers. AB - Oral treatment with chitosan with a molecular weight approximately 105, but not its oligomer, reduced plasma content of free-radical oxidation products in normal rats and animals treated for the bone marrow form of radiation sickness and stimulated the recovery processes in involved bone marrow and peripheral blood. PMID- 17415438 TI - Distribution and elimination of milk angiogenin in mouse immunocompetent organs and brain. AB - Dynamics of angiogenin content in the serum, thymus, bone marrow, and brain of mice was studied after intravenous injection. The maximum angiogenin retention was detected in the thymus, while high rate of its elimination in the brain. PMID- 17415439 TI - Gastroprotective properties of 11-deoxymisoprostol (prostaglandin E1 analog) and its effect on the level of sialic acids in gastric tissue of rats with peptic ulcer. AB - 11-Deoxymisoprostol (prostaglandin E1 analog) exhibited a pronounced gastroprotective effect on various models of experimental ulcers induced by nonsteroid antiinflammatory drugs. A relationship between high resistance of the gastroduodenal mucosa under the effect of 11-deoxymisoprostol and changes in the level of sialic acid was detected. PMID- 17415440 TI - In vivo anticancer activity of lysates from trypanosoma cruzi of different genetic groups. AB - Lyzed epimastigotes of Trypanosoma cruzi clones P209-1, Gamba1, Sp104-1, MASu, Y7/1, MN12, Cl-Brener, 86/2036, Y7/2-1 inhibit the growth of Ehrlich adenocarcinoma in mice. the tumor decreased 1.5-3 times after 12 daily injections of lysates from 15 million epimastigotes. The protective effect progressed after the injections were discontinued and depended on the dose and lysate producer clone. Trypanosoma lysates in the studied doses were nontoxic. PMID- 17415441 TI - Development of differential sensitivity of CaOv ovarian adenocarcinoma cells to antitumor agents under conditions of hypoxia. AB - We studied the role of VEGF signal pathway in autocrine regulation of tumor cell growth and survival under conditions of hypoxia. Hypoxia-resistant CaOv/H substrain with high level of VEGF-A secretion was obtained by long-term culturing of CaOv ovarian adenocarcinoma cells with CoCl2 (hypoxia inductor). VEGF-A directly participates in autocrine regulation of CaOv cell growth, including the maintenance of cell growth under conditions of hypoxia or cytostatic treatment. On the other hand, CaOv/H cells retain high apoptotic potential and are characterized by high expression of p27Kip1 (cyclin-dependent kinase inhibitor), which attests to possible involvement of this inhibitor into the regulation of apoptotic response of cells under conditions of hypoxia. PMID- 17415442 TI - Synthesis of highly tritium labeled steroids for evaluation of aromatase and steroidsulfatase activities in endometrial tumors. AB - Data on activities of estrogen-synthesis enzymes (aromatase and steroidsulfatase) in endometrial tumors were obtained by sensitive and highly precise radioisotope methods. The results clear out some pathogenetic aspects in the development and progress of malignant tumors in the endometrium. PMID- 17415443 TI - Study of optical parameters of the nucleoli under the effect of transcription inhibitors by coherent phase microscopy. AB - The use of coherent phase microscopy for online quantitative registration of nucleolar reaction to transcription inhibition is validated. Reduction of phase thickness of the nucleoli was detected during the first minutes of the experiment; 30 min after addition of the drug rarefaction zones predominated and areas of condensation were seen. These changes reflect the dynamics of disorders in the nucleolar ultrastructure during transcription inhibition. PMID- 17415444 TI - Roller organ cultures of the retina from postnatal rats. AB - Whole retinas of 2-14-day-old rats were cultured in a roller device for 2-14 days. Floating retinas of 7-14-day-old rats formed hole spheroid structures (spheroids) with the wall completely retaining the linear structure and layer-by layer cellular and fibrous architecture, including the outer nuclear, outer plexiform, inner nuclear, inner plexiform layers, layers of ganglion cells and nerve fibers. The retina obtained at earlier terms of development often formed folds, with pyknotic nuclei of dead neurons in their deep compartments. In organ cultures of the retina isolated from rats at early postnatal periods, rosettes were formed in sites of local injury to the outer nuclear layer and pigmented epithelium. Roller organ cultures can be used for in vitro studies of the development and experimental diseases of the retina. PMID- 17415445 TI - Proliferative activity of retinal vascular cells in newborn rat at different oxygenation modes. AB - We evaluated the relationship between the percentage of oxygen in inhaled air and alternation of this parameter and proliferative activity of cells in retinal vessels of normal newborn rats. The relationships between these parameters and the mean diameter of retinal vessels were evaluated. The study was carried out on total retinal preparations and tangential sections of the retina by the immunoperoxidase and immunofluorescent methods. Hypoxia and hyperoxia significantly suppressed proliferative activity, while alternation of hyperoxia and normoxia significantly increased both proliferative activity of vascular cells and the mean diameter of retinal vessels. PMID- 17415446 TI - Changes in wound field lipids in rat skin. AB - Intricate dynamics of lipid transformations was detected in rat skin wound field during regeneration. The content of lipids increased in the granulation fibrous tissue during the early period of regeneration and decreased later. On day 23 the regeneration process was over, the formed scar tissue differed from intact skin by the content of many lipid fractions. in the crust and skin sites adjacent to the wound the regeneration was associated with significant changes in the lipid composition. PMID- 17415447 TI - Multipotent stem and progenitor cells of the olfactory epithelium. AB - In recent decades, a wide spectrum of fetal and embryonic stem and progenitor cells were used for cell therapy of diseases of the central nervous system, but the olfactory glial ensheathing cells exhibited certain advantages due to their biological properties and capacity to stimulate regeneratory processes in spinal injury. The therapeutic effect of a heterogeneous complex of olfactory epithelial cells is more pronounced; apart from glial ensheathing cells, this complex includes fibroblasts, Schwann cells, stem and progenitor cells of this structure. The use of minimally invasive methods for isolation of human olfactory epithelial tissue is important for clinical practice, because they provide cells for autologous transplantation and rule out graft rejection immune reaction and the risk of transmission viral infection and transfer of genetic defects, which can be associated with allotransplantation. PMID- 17415448 TI - In vitro and in vivo differentiation of mesenchymal stem cells in the cardiomyocyte direction. AB - The possibility of mesenchymal stem cell differentiation in the cardiomyocyte direction was studied on Wistar-Kyoto rats with myocardial infarction induced by ligation of the left coronary artery. In vitro treatment of mesenchymal stem cells with 5-azacitidine led to spontaneous contractions of about 15% cells in culture. Analysis of the expression of matrix RNA showed expression of fetal and functional markers of the myocardium in this cell culture. In vivo on day 21 after myocardial infarction and intravenous transplantation of mesenchymal stem cells into the periinfarction area, myocardial cells carrying donor label were detected. Immunohistochemical analysis showed that these cells were cardiomyocytes integrated into the myocardium. These cells can be a result of differentiation of transplanted mesenchymal stem cells or fusion of endogenous cardiomyocytes with exogenous mesenchymal stem cells. PMID- 17415449 TI - Combined immunoperoxidase analysis for visualization of cells of the blood-brain barrier. AB - A method for double immunoperoxidase staining of the blood-brain barrier cell elements was developed using 2mB6 monoclonal antibodies specifically visualizing brain capillary antigen and antibodies to glial fibrillary acidic protein (GFAP). The method is based on consecutive visualization of antigen structures in one section: first using a substrate mixture containing diaminobenzidine (brown coloring) and then a mixture with diaminobenzidine and CoCl(2) (blue coloring). This method visualizes cerebral capillary cells and fibrillar astrocytes interacting with them. The method can be used in all variants of double immunoperoxidase studies and for immunodetection of several antigens in immunoblotting analysis. PMID- 17415450 TI - Effect of mesenchymal stem cell transplantation on cognitive functions in rats with ischemic stroke. AB - The effect of intravenous transplantation of mesenchymal stem cells on the recovery of cognitive functions was studied in Wistar-Kyoto rats after brain stroke induced by occlusion of the middle cerebral artery in the left hemisphere. Analysis 2 and 5 weeks after stroke showed that transplantation of mesenchymal stem cells 3 days after middle cerebral artery occlusion reduced the area of cerebral injury, preserved cognitive functions, and decreased mortality in experimental animals. PMID- 17415451 TI - Optimum conditions for culturing of human bone marrow and adipose tissue mesenchymal precursor cells. AB - We present a technology of culturing of human mesenchymal stem cells under conditions excluding the presence of animal sera or additional growth factors, but preserving high proliferative potential and the capacity of these cells to multilineage differentiation. Human umbilical serum was used as the alternative material. We found that in the presence of human umbilical serum mesenchymal stem cells more effectively proliferate and retain their differentiation capacity. The proposed technology yields 109-1010 morphologically and functionally identical cells. PMID- 17415452 TI - Cytofluorometric analysis of phenotypes of human bone marrow and umbilical fibroblast-like cells. AB - Comparative analysis of the expression of some surface markers of human bone marrow mesenchymal stem cells, umbilical fibroblast-like cells, and skin fibroblasts was carried out by the flow cytofluorometry method. Mesenchymal stem cells and umbilical fibroblast-like cells were similar by the levels of expression of the main histocompatibility complex antigens, adhesion molecules, and some growth factor receptors. The profile of skin fibroblast surface antigens was characterized by higher expression of the markers typical of differentiated cells. The results prove the possibility of using umbilical fibroblast-like cells as an alternative source of mesenchymal stem cells for cell replacement therapy. PMID- 17415453 TI - Stromal regulation of hemopoietic stem cells in long-term human bone marrow tissue cultures under the effect of parathyroid hormone. AB - We studied the interaction between different categories of hemopoietic precursors with parathyroid hormone-activated stromal microenvironment. Improved survival of early precursors capable long-term hemopoiesis maintenance and increased number of later short-term repopulating precursors was demonstrated on the model of co culturing of human bone marrow cells on a layer of adherent cells of long-term bone marrow cultures treated with parathyroid hormone. These changes correlate with increased expression of genes involved in the maintenance of the hemopoietic stem cells in the sublayer activated by parathyroid hormone. Simultaneously, the expression of some stromal differentiation genes, adhesion molecules for hemopoietic stem cells, and growth factors increased in adherent cell layers treated with parathyroid hormone. These findings attest to activating effect of parathyroid hormone on cells forming the niches for both early and later hemopoietic precursors, and hence parathyroid hormone can be used as a potential agent promoting expansion of early hemopoietic stem cells ex vivo. PMID- 17415454 TI - The role of donor and recipient in tissue implantation. AB - The relationships between embryonic stem and cambial cells in the ontogeny were evaluated on the basis of our data on organ embryogenesis and in vivo implantation of epithelial tissues and published data. We demonstrated the role of recipient tissues in the implantation process. Aseptic inflammation developing in response to the implant activates proliferation of the adjacent donor tissues. Proliferation and differentiation of the implanted donor tissues correspond to inflammation phases in the focus of implantation, are regulated by factors of the recipient organism, and are histogenetically determined. PMID- 17415455 TI - Effect of mesenchymal stem cells on rejection of xenogenic bone transplant. AB - We studied the effects of mesenchymal stem cells on rejection of xenogenic bone transplant. Experiments were carried out on Wistar-Kyoto rats transplanted chicken demineralized bone matrix populated or not populated with mesenchymal stem cells to the site of parietal bone defect. Histological analysis showed complete resorption of the xenogenic transplant not populated with mesenchymal stem cells by day 119. In experimental animals chicken matrix without signs of tissue inflammation was in fact completely retained over the entire period of observation. Numerous new vessels, mineralization fields, and areas of bone tissue formation were seen in the transplant. PMID- 17415456 TI - Use of thermosensitive polymer material on the basis of N-isopropylacrylamide and N-tert-butylacrylamide copolymer in cell technologies. AB - We developed thermosensitive polymer substrates on the basis of N isopropylacrylamide and N-tert-butylacrylamide co-polymer and studied their interaction with cultured substrate-dependent mammalian cells. It was shown that these polymers promote cell adhesion and proliferation at a level comparable to polystyrene treated for cell culturing and provide effective cell detachment after lowering culturing temperature below a critical level determined by phase transition temperature in aqueous solutions of polymers. A dependence of phase transition temperature on the ratio between N-isopropylacrylamide and N-tert butylacrylamide was demonstrated. Differences in the dynamics of cell detachment from the surface of polymer substrates with various proportions between the components were shown. PMID- 17415457 TI - Effect of hypobaric hypoxia on the development of long-term posttetanic potentiation in slices of rat olfactory cortex: correction with hypoxic preconditioning. AB - We studied the effect of in vivo hypobaric hypoxia on the development (after 3 h) of in vitro long-term posttetanic potentiation in Wistar rats. Severe hypoxia suppressed induction of posttetanic potentiation in slices of the olfactory cortex. Preconditioning exposure (moderate hypobaric hypoxia) prevented inhibition of posttetanic potentiation induced by severe hypoxia. PMID- 17415458 TI - Dependence of blood pressure changes in cats during acute hypoxic hypoxia on the type of carotid sinus reflex. AB - The type and degree of blood pressure changes in cats were studied in acute experiments under conditions of acute hypoxic hypoxia (40% decrease in oxygen partial pressure). During hypoxia, blood pressure increased in cats with pressor type of the carotid sinus reflex and decreased in animals with depressor type of this reflex. Our results indicate that the direction and degree of hypoxic changes in blood pressure in animals coincide with variations in this parameter in response to the carotid sinus reflex. PMID- 17415459 TI - Role of hepatic opioid receptors in the regulation of bile excretion. AB - Experiments on isolated rat liver perfused with Ringer-Krebs bicarbonate buffer showed that stimulation of delta-opiate receptors in this organ increases bile flow rate and taurocholate secretion. Stimulation of micro-opiate receptors decelerated bile production and inhibited taurocholate secretion. Acceleration of bile production and stimulation of taurocholate secretion under the influence of dalargin is probably related to its interaction with delta-opiate receptors in the liver. PMID- 17415460 TI - Activities of prolyl endopeptidase and dipeptidyl peptidase IV in brain structures of rats with dopamine deficiency-dependent MPTP-induced depressive syndrome. AB - The development of MPTP-induced depressive syndrome in rats was accompanied by activation of prolyl endopeptidase and dipeptidyl peptidase IV in the brain frontal cortex. Prolyl endopeptidase activity in the striatum also increased under these conditions. Our results indicate that proline-specific peptidases in the target structures of the brain dopaminergic system are involved in the pathogenesis of dopamine deficiency-dependent depressive states. PMID- 17415461 TI - Role of cannabinoid receptors in the regulation of cardiac contractility during ischemia/reperfusion. AB - We studied the effect of selective ligands of cannabinoid (CB) receptors on contractility of isolated Langendorff-perfused rat heart under conditions of 45 min total ischemia and 30-min reperfusion. Perfusion with a solution containing selective CB receptor agonist HU-210 for 10 min before ischemia increased the severity of reperfusion contractile dysfunction. This drug decreased left ventricular developed pressure and maximum rates of contraction and relaxation, but had no effect on heart rate and end-diastolic pressure. The negative inotropic effect of the drug was transitory and disappeared after 5-min reperfusion. Pretreatment with selective CB1 receptor antagonist SR141716A and selective CB2 receptor antagonist SR144528 had no effect on heart rate and myocardial contractility during reperfusion. Our results indicate that stimulation of CB receptors can increase the degree of reperfusion-induced cardiac contractile dysfunction. However, endogenous cannabinoids are not involved in the development of myocardial contractile dysfunction during ischemia/reperfusion of the isolated heart. PMID- 17415462 TI - Neuroimmunomodulatory effect of antibodies against GABA on acute generalized and chronic epileptiform activity. AB - We studied the possibility of induction of anti-GABA autoantibodies during chronic epileptization of the brain and the effect of systemic intraperitoneal administration of anti-GABA antibodies to C57Bl/6 mice on acute generalized and chronic epileptiform activity. It was found for the first time that chronic epileptization is accompanied by induction of anti-GABA autoantibody synthesis. Antibodies against GABA produced a proepileptic effect. PMID- 17415463 TI - Mechanisms of hemolytic anemia during experimental methemoglobinemias. AB - A complex study of the peripheral erythron component was performed during methemoglobinemias induced by single administration of sodium nitrate and phenylhydrazine in LD50. Administration of methemoglobin-forming agents to rats induced the development of hemolytic anemia. The pathogenesis of this disorder included significant long-term modifications of the erythrocyte membrane. The severity and duration of anemia syndrome depended on chemical structure of xenobiotics, blood methemoglobin level, and the duration of the acute period of methemoglobinemia. PMID- 17415464 TI - Relationship between natural concentration of heavy water isotopologs and rate of H2O2 generation by mitochondria. AB - Relationship between natural concentration of heavy water isotope homologs in natural water and the kinetics of H(2)O(2) generation by isolated rat liver mitochondria in the presence of succinic acid as the substrate was studied. Natural concentrations of heavy water isotopologs significantly inhibited the studied reaction. The decrease in heavy isotopolog content in natural water leads to disinhibition and significant acceleration of the studied reaction. PMID- 17415465 TI - Cell microvesicles during experimental endotoxemia. AB - The dynamics of microvesicle formation in arterial blood in generalized Schwartzman phenomenon was studied. Successive (with 24-h interval) intravenous injections of endotoxin to rabbits in a dose of 1 mg/kg and 3 mg/kg caused an increase in the content of microvesicles in the blood, some of them containing ecto-5'-nucleotidase. Biphasic changes in arterial blood clotting time and erythrocyte hemolysis were observed. PMID- 17415466 TI - Plasma content of cholesterol and glycerol alcohols depends on the number of fatty acid double bonds in lipoprotein lipid pool. AB - The number of the fatty acid double bonds (unsaturation degree) in the plasma lipid pool was evaluated by automated ozone titration. A positive relationship between the content of double bonds, cholesterol, and glycerol was detected. The higher was plasma cholesterol level, the more double bonds contained fatty acids. At cholesterol level of 0.8-18.9 mmol/liter the double bond/cholesterol ratio approached 4.0 and double bond/glycerol ratio was 1.3-1.5. The maximum content of double bonds was detected in LDL. It was hypothesized that the greater part of plasma cholesterol is esterified with arachidonic acid possessing 4 double bonds; three fatty acids are esterified with glycerol, which altogether have 1-3 double bonds. It seems that plasma cholesterol level indirectly but significantly reflects the level of essential polyenic fatty acids, while glycerol reflects the levels of saturated and unsaturated fatty acids. The role of cholesterol in the plasma consists in the participation in transfer of essential fatty acid in the nonpolar form and their absorption by cells. LDL are the main transmitters of essential polyenic fatty acids in the form of cholesterol esters. PMID- 17415467 TI - Effect of serum from cardiovascular patients on catalytic activity of secretory phospholipase A2 (IIA). AB - Incubation of patients' serum catalytically active by type IIA secretory phospholipase A2 (SP-IIA) with serum containing the enzyme in a high concentration but exhibiting no catalytic activity in 1:1 volume ratio led to a significant inhibition of SP-IIA catalytic activity. donor and patient sera with low levels of SP-IIA had no effect on the serum with SP-IIA activity under these conditions. However, the increase in the content of patients' serum with a low level of SP-IIA in the incubation mixture to 1:2 (v/v) and of donor serum to 1:3 (v/v) also led to blockade of SP-IIA catalytic activity. These results indicate that human serum contains an SP-IIA inhibitor and its concentration decreases significantly in sera with SP-IIA activity. PMID- 17415468 TI - Adaptive effects of dexamethasone in stress exposure. AB - Disorders in the compensatory adaptive processes underlie the development of many somatic diseases. The neuroimmunoendocrine complex and one of its main components, the hypothalamic-pituitary-adrenal system, play the leading role in the maintenance of the adaptation reserve. Treatment with a synthetic analog of hydrocortisone (final effector of this system) in low doses modulates its function by inducing positive changes in the adaptive mechanisms under conditions of stress exposure. PMID- 17415469 TI - Comparative analysis of efficiency and specificity of various sorbents for apheresis of low-density lipoproteins. AB - Efficiency of sorbents for LDL apheresis was compared in vitro. The sorbents based on ion-exchange interaction of the ligand with LDL (Liposorber, DALI) exhibited minimum specificity towards the eliminated component, while immunosorbents (LNP-Lipopak, LDR-TheraSorb) were most efficient. By sorption capacity, the available hemosorbents are inferior to plasmasorbents, which explains low efficiency of the therapy based on single application of hemosorbents especially in patients with consederably increased content of LDL cholesterol. PMID- 17415470 TI - Pharmacodynamic parameters of anticoagulants based on sulfated polysaccharides from marine algae. AB - Fucoidans isolated from Fucus evanescens and Laminaria cichorioides kelp can inhibit thrombin and factor Xa of the blood coagulation system. In rats, intravenous injection of fucoidans dose-dependently increased anticoagulant activity of the plasma. Fucoidans can form complexes with protamine sulfate. The observed quantitative differences in the action of fucoidans can result from different sulfation degree and the presence of various types of glycoside bonds in polysaccharide molecules. PMID- 17415471 TI - Effect of 5HT2C-receptor agonist MK-212 on blood corticosterone level and behavior in mice. AB - A dose-dependent the effect of 5HT2C-receptor agonist MK-212 on mouse behavior was demonstrated. Intraperitoneal injection of MK-212 in high doses (0.5 and 1.0 mg/kg) increased blood level of corticosterone in mice and reduced their motor activity. In low doses of 0.1 and 0.2 mg/kg, the agonist reduced anxiety, but had no effect on motor activity. It is hypothesized that low doses of MK-212 exhibited anxiolytic activity in mice. PMID- 17415472 TI - Naloxone-blocked depriming effect of anxiolytic selank on apomorphine-induced behavioral manifestations of hyperfunction of dopamine system. AB - Peptide anxiolytic selank (Thr-Lys-Pro-Arg-Pro-Gly-Pro) applied intraperitoneally in doses of 0.01, 0.1, 1.0, and 10.0 mg/kg to mice reduces behavioral manifestations of dopaminergic system induced by apomorphine in the verticalization test. This effect was comparable to that of atypical antipsychotic olanzapine in near-therapeutic doses (0.1 and 1.0 mg/kg, intraperitoneally) and was blocked with nonselective opioid receptor antagonist naloxone (10 mg/kg, intraperitoneally). Radioreceptor assay showed that selank did not displace nonselective D2-dopamine receptor antagonist (3)H-spiperone (EC50>100 microM) and delta- and micro-opioid receptor ligand 3H-DADLE (EC50>40 microM) from specific binding sites on rat brain membranes. It is hypothesized that the revealed behavioral effect of selank is mediated by its modulating effect on the endogenous opioid system and specifically, by its effect on activity of enkephalin-degrading enzymes. PMID- 17415473 TI - Effect of chronic consumption of sodium valproate and melatonin on seizure activity in Krushinskii-Molodkina rats. AB - Experiments on Krushinskii-Molodkina rats with hereditary predisposition to audiogenic seizures showed that chronic consumption of aqueous solution of melatonin (50 mg/liter) had no effect on the pattern of seizures induced by 20 fold acoustic stimulation. Sodium valproate (50 mg/liter) insignificantly decreased the seizure response. Combined treatment with sodium valproate and melatonin produced a potent anticonvulsant effect, i.e. increased the latency and decreased the severity of audiogenic seizures. However, myoclonus in animals receiving combined treatment with these drugs developed much more rapidly compared to rats receiving melatonin or sodium valproate monotherapy. PMID- 17415474 TI - Circadian variations in immune values and serum melatonin in asthmatics. AB - Circadian variations in the immune status and serum melatonin were studied in asthmatics during exacerbation stage before and after treatment. Deviations from the normal (donor) immunogram values at 9.00 and 21.00 and decreased correlations between immune values and blood melatonin level were detected. The correlations of immune values between each other and with melatonin over the circadian cycle increased after treatment. PMID- 17415475 TI - Surface adhesins of lactobacilli loosely connected to the cell wall and eliminated into the environment during culturing in liquid nutrient media. AB - Six lactobacillus species and 4 clones of one of them were studied in order to clear out the ratio between the adhesion capacities of concanavalin A-reactive glycoprotein adhesins on the surface of the bacterial cell and glycoprotein adhesins released into the broth during culturing in liquid nutrient media. The adhesive activity of cultures is largely determined by the strain rather than species appurtenance. Elimination of glycoprotein adhesins from the bacterial cell and their antagonistic activity towards Candida albicans were demonstrated in specific interactions of glycoprotein adhesins with immune serum and concanavalin A. PMID- 17415476 TI - Plasma NO-binding activity in patients with hematological malignancies: suppressive effect of glucocorticoids. AB - Plasma NO-binding activity was studied in patients with various forms of hematological malignancies. The method used in the study quantitatively evaluated the plasma capacity to bind NO, which reflects the intensity of NO production and the degree of patient's stress resistance. Plasma NO-binding activity significantly decreases in patients with hematological malignancies. Glucocorticoid treatment promotes the decrease in plasma NO-binding activity, which was dose-dependent. PMID- 17415477 TI - Different effects of carbon ion and gamma-irradiation on expression of receptor activator of NF-kB ligand in MC3T3-E1 osteoblast cells. AB - We investigated the effects of carbon ion and gamma-irradiation on osteoblastic MC3T3-E1 cells by comparing mRNA expression levels for RANKL and osteoprotegerin by RT-PCR. MC3T3-E1 cells were irradiated with 2, 4, or 6 Gy of carbon ions or gamma-rays, and total RNA was harvested 1, 2, 3, 5, or 7 days after irradiation. The RANKL mRNA/OPG mRNA ratio in carbon ion-irradiated MC3T3-E1 cells was lower, while in gamma-irradiated MC3T3-E1 cells this ratio was higher than in non irradiated cells. To evaluate osteoclastogenesis of MC3T3-E1 cells, carbon ion- or gamma-irradiated cells were co-cultured with non-irradiated cells from murine bone marrow. Staining for tartrate-resistant acid phosphatase (TRAP) in co cultures showed that carbon ion irradiation suppressed osteoclastogenesis. This result is consistent with the lower RANKL/OPG mRNA ratio for carbon ion irradiated cells. These results suggest that carbon ion irradiation acts primarily on osteoblastic cells, leading to a decrease in the RANKL/OPG mRNA ratio. This effect, in turn, leads to a decrease in osteoclastogenesis and osteoclast activity, which results in an increase in bone volume. PMID- 17415478 TI - Possible involvement of lactate in neuroglial interaction through nicotinic cholinergic synapses in the cranial cervical sympathetic ganglion. AB - Activities of LDH and its H- and M-isoforms in neurons and satellite gliocytes of the cranial cervical sympathetic ganglion in rabbits under normal conditions and during nicotinic cholinergic synapse blockade were evaluated by integral cytophotometry in tissue sections. Normally activity of H-isoform predominates in neurons and M-isoform in satellite gliocytes. Blockade of the cranial cervical sympathetic ganglion significantly decreased LDH activity (H- and M-isoforms) in neurons in direct proportion to the number of blocked nicotinic cholinergic receptors. Activity of M-isoform in satellite gliocytes decreased with increasing the degree of blockade, while activity of H-isoform did not change. The isoenzyme profile of LDH in satellite gliocytes reached the level of intact neurons. Presumably, lactate production in satellite gliocytes is regulated by sympathetic neurons through nicotinic cholinergic synapses. PMID- 17415479 TI - Morphological and immunological analysis of the oral mucosa in tobacco smoking and odontopreparation. AB - Tobacco smoking causes pronounced structural reorganization of the gingival mucosa and its atrophic changes. Odontopreparation stimulates destructive processes in the gingival mucosa promoting activation of local immunity (stimulation of immunoglobulin synthesis). Significant differences in the local humoral immunity (IgG dysimmunoglobulinemia) and cytokine spectrum in nonsmoking and tobacco-smoking patients were detected. Subcompensated level of immune resistance in tobacco smokers prompts referring them to a group at a high risk of chronic pathological processes of the oral cavity. PMID- 17415480 TI - Structural reactions of the buccal mucosa in diabetic parodontopathy. AB - Pronounced changes in the capillaries, hemodynamic disorders, epitheliocyte degeneration and atrophy develop in the buccal mucosa of patients with types I and II diabetes mellitus in the absence of inflammatory cellular infiltration or with facultative infiltration. The morphogenesis of pathological changes can be regarded as primary diabetic microangiopathy causing metabolic disorders with the development of degenerative and atrophic changes in all structural components of the buccal mucosa and development of diabetic parodontopathy, a primary degenerative process. PMID- 17415481 TI - A method for evaluating the fluctuations in the intensity of substance transport between blood and bones in the predominant direction. AB - The dynamics of fluctuations in the intensity of the blood-bone flow of substances in the predominant and opposite directions was evaluated using the percent radioactivity difference coefficient and its graphic presentation. The technology of calculating this coefficient is described. PMID- 17415482 TI - Rapid method for evaluation of cholinesterase inhibitors. AB - The fluorescence intensity of reversible inhibitor ethidium bromide fluorophore complex with equine blood butyryl cholinesterase decreases in the presence of inhibitor (tacrine) not fluorescing in the visible spectrum. An express method for tacrine evaluation is developed. PMID- 17415483 TI - The use of statistical methods for predicting the status of patients with acute cerebral and coronary ischemia. AB - The most significant informative clinical laboratory parameters were determined and the exceptional value of cryoglobulin concentrations for predicting the clinical status was demonstrated. The efficiency of the proposed protocol of selective plasmapheresis for patients with atherothrombotic variant of stroke is confirmed, mathematical models describing and predicting the patient status are plotted. PMID- 17415484 TI - Model of immunogenic uveitis in rabbits. AB - Six models of experimental immunogenic uveitis induced by injection of normal horse serum were tried in rabbits. A convenient working variant of the model was developed characterized by high activity and long duration of the inflammatory process in the eye. Creation of this model requires pre-sensitization (subcutaneous injection of 5 ml normal horse serum) followed by intravitreal injection of the resolving dose of horse serum on day 9. An adapted improved model of experimental immunogenic uveitis in rabbits is proposed. PMID- 17415485 TI - Effect of hypoxia during early organogenesis on cardiac activity and noradrenergic regulation in the postnatal period. AB - Cardiac activity in rats during the postnatal period was studied in vitro and in vivo after exposure of rat pups to antenatal acute hypobaric hypoxia at the stage of organogenesis (day 9-10 of gestation). Cultured cardiomyocytes from rat pups exposed to antenatal hypoxia were characterized by increased rate of contractions and decreased reactivity to norepinephrine. Heart rate elevation, predominance of sympathetic influences on cardiac activity, and significant increase in norepinephrine concentration in the cerebral cortex were found in freely moving animals exposed to antenatal hypoxia. Our results indicate that hypoxia at the stage of organogenesis modulated cardiac activity during the postnatal period, which manifested at the level of effector structures in the heart and activity of regulatory systems. PMID- 17415487 TI - Intermediate temperature proton conductors for PEM fuel cells based on phosphonic acid as protogenic group: a progress report. AB - The melting behaviour and transport properties of straight chain alkanes mono- and difunctionalized with phosphonic acid groups have been investigated as a function of their length. The increase of melting temperature and decrease of proton conductivity with increasing chain length is suggested to be the consequence of an increasing ordering of the alkane segments which constrains the free aggregation of the phosphonic acid groups. However, the proton mobility is reduced to a greater extent than the proton diffusion coefficient indicating an increasing cooperativity of proton transport with increasing length of the alkane segment. The results clearly indicate that the "spacer concept", which had been proven successful in the optimization of the proton conductivity of heterocycle based systems, fails in the case of phosphonic acid functionalized polymers. Instead, a very high concentration of phosphonic acid functional groups forming "bulky" hydrogen bonded aggregates is suggested to be essential for obtaining very high proton conductivity. Aggregation is also suggested to reduce condensation reactions generally observed in phosphonic acid containing systems. On the basis of this understanding, the proton conductivities of poly(vinyl phosphonic acid) and poly(meta-phenylene phosphonic acid) are discussed. Though both polymers exhibit a substantial concentration of phosphonic acid groups, aggregation seems to be constrained to such an extent that intrinsic proton conductivity is limited to values below sigma = 10(-3) S cm(-1) at T = 150 degrees C. The results suggest that different immobilization concepts have to be developed in order to minimize the conductivity reduction compared to the very high intrinsic proton conductivity of neat phosphonic acid under quasi dry conditions. In the presence of high water activities, however, (as usually present in PEM fuel cells) the very high ion exchange capacities (IEC) possible for phosphonic acid functionalized ionomers (IEC >10 meq g(-1)) may allow for high proton conductivities in the intermediate temperature range (T approximately 120 -160 degrees C). PMID- 17415488 TI - Carbon materials for supercapacitor application. AB - The most commonly used electrode materials for electrochemical capacitors are activated carbons, because they are commercially available and cheap, and they can be produced with large specific surface area. However, only the electrochemically available surface area is useful for charging the electrical double layer (EDL). The EDL formation is especially efficient in carbon pores of size below 1 nm because of the lack of space charge and a good attraction of ions along the pore walls. The pore size should ideally match the size of the ions. However, for good dynamic charge propagation, some small mesopores are useful. An asymmetric configuration, where the positive and negative electrodes are constructed from different materials, e.g., activated carbon, transition metal oxide or conducting polymer, is of great interest because of an important extension of the operating voltage. In such a case, the energy as well as power is greatly increased. It appears that nanotubes are a perfect conducting additive and/or support for materials with pseudocapacitance properties, e.g. MnO(2), conducting polymers. Substitutional heteroatoms in the carbon network (nitrogen, oxygen) are a promising way to enhance the capacitance. Carbons obtained by one step pyrolysis of organic precursors rich in heteroatoms (nitrogen and/or oxygen) are very interesting, because they are denser than activated carbons. The application of a novel type of electrolyte with a broad voltage window (ionic liquids) is considered, but the stability of this new generation of electrolyte during long term cycling of capacitors is not yet confirmed. PMID- 17415489 TI - Hydrogen storage in nanoporous carbon materials: myth and facts. AB - We used Grand canonical Monte Carlo simulation to model the hydrogen storage in the primitive, gyroid, diamond, and quasi-periodic icosahedral nanoporous carbon materials and in carbon nanotubes. We found that none of the investigated nanoporous carbon materials satisfy the US Department of Energy goal of volumetric density and mass storage for automotive application (6 wt% and 45 kg H(2) m(-3)) at considered storage condition. Our calculations indicate that quasi periodic icosahedral nanoporous carbon material can reach the 6 wt% at 3.8 MPa and 77 K, but the volumetric density does not exceed 24 kg H(2) m(-3). The bundle of single-walled carbon nanotubes can store only up to 4.5 wt%, but with high volumetric density of 42 kg H(2) m(-3). All investigated nanoporous carbon materials are not effective against compression above 20 MPa at 77 K because the adsorbed density approaches the density of the bulk fluid. It follows from this work that geometry of carbon surfaces can enhance the storage capacity only to a limited extent. Only a combination of the most effective structure with appropriate additives (metals) can provide an efficient storage medium for hydrogen in the quest for a source of "clean" energy. PMID- 17415490 TI - Fructose/dioxygen biofuel cell based on direct electron transfer-type bioelectrocatalysis. AB - One-compartment biofuel cells without separators have been constructed, in which d-fructose dehydrogenase (FDH) from Gluconobacter sp. and laccase from Trametes sp. (TsLAC) work as catalysts of direct electron transfer (DET)-type bioelectrocatalysis in the two-electron oxidation of d-fructose and four-electron reduction of dioxygen as fuels, respectively. FDH adsorbs strongly and stably on Ketjen black (KB) particles that have been modified on carbon papers (CP) and produces the catalytic current with the maximum density of about 4 mA cm(-2) without mediators at pH 5. The catalytic wave of the d-fructose oxidation is controlled by the enzyme kinetics. The location and the shape of the catalytic waves suggest strongly that the electron is directly transferred to the KB particles from the heme c site in FDH, of which the formal potential has been determined to be 39 mV vs. Ag|AgCl|sat. KCl. Electrochemistry of three kinds of multi-copper oxidases has also been investigated and TsLAC has been selected as the best one of the DET-type bioelectrocatalyst for the four-electron reduction of dioxygen in view of the thermodynamics and kinetics at pH 5. In the DET-type bioelectrocatalysis, the electron from electrodes seems to be transferred to the type I copper site of multi-copper oxidases. TsLAC adsorbed on carbon aerogel (CG) particles with an average pore size of 22 nm, that have been modified on CP electrodes, produces the catalytic reduction current of dioxygen with a density of about 4 mA cm(-2), which is governed by the mass transfer of the dissolved dioxygen. The FDH-adsorbed KB-modified CP electrodes and the TsLAC-adsorbed CG modified CP electrodes have been combined to construct one-compartment biofuel cells without separators. The open-circuit voltage was 790 mV. The maximum current density of 2.8 mA cm(-2) and the maximum power density of 850 microW cm( 2) have been achieved at 410 mV of the cell voltage under stirring. PMID- 17415491 TI - The potential of organic polymer-based hydrogen storage materials. AB - The challenge of storing hydrogen at high volumetric and gravimetric density for automotive applications has prompted investigations into the potential of cryo adsorption on the internal surface area of microporous organic polymers. A range of Polymers of Intrinsic Microporosity (PIMs) has been studied, the best PIM to date (a network-PIM incorporating a triptycene subunit) taking up 2.7% H(2) by mass at 10 bar/77 K. HyperCrosslinked Polymers (HCPs) also show promising performance as H(2) storage materials, particularly at pressures >10 bar. The N(2) and H(2) adsorption behaviour at 77 K of six PIMs and a HCP are compared. Surface areas based on Langmuir plots of H(2) adsorption at high pressure are shown to provide a useful guide to hydrogen capacity, but Langmuir plots based on low pressure data underestimate the potential H(2) uptake. The micropore distribution influences the form of the H(2) isotherm, a higher concentration of ultramicropores (pore size <0.7 nm) being associated with enhanced low pressure adsorption. PMID- 17415492 TI - Direct electron transfer in nanostructured sol-gel electrodes containing bilirubin oxidase. AB - Bilirubin oxidase encapsulated within a silica sol-gel/carbon nanotube composite electrode effectively catalyzed the reduction of molecular oxygen into water through direct electron transfer at the carbon nanotube electrode surface. In this nanocomposite approach, the silica matrix is designed to be sufficiently porous for substrate molecules to have access to the enzyme and yet provides a protective cage for immobilization without affecting biological activity. The incorporation of carbon nanotubes adds electrical connectivity and increases active electrode surface area. The standard surface electron transfer rate constant was calculated to be 59 s(-1) which indicates that the carbon nanotube side walls are primarily responsible for electron transfer. The use of direct electron transfer processes simplifies biofuel cell fabrication by eliminating the need for redox mediator and ion-conducting separators. PMID- 17415493 TI - Electrochemical kinetics of porous, carbon-decorated LiFePO4 cathodes: separation of wiring effects from solid state diffusion. AB - We try to identify the rate-determining step of electrochemical kinetics of a LiFePO(4)-carbon composite electrode by varying the mass of electrode and, additionally, by varying the charge-discharge current in a wide range. It is shown that the reversible capacity is almost independent of electrode mass at currents lower than ca. 1 C (170 mAh g(-1)). At higher currents, however, the reversible capacity starts to drop significantly. The electrode resistance determined from the corresponding polarization voltage shows inverse proportionality with mass at currents smaller than 1 C. At higher currents the electrode resistance is almost independent of electrode mass. We conclude that at lower currents (below 1 C) the main transport step is related to the active particles themselves (either to incorporation reaction or solid state diffusion of Li). At higher currents the contribution of electronic and ionic transport towards the active particles becomes substantial and should be taken into account when designing high-rate insertion electrodes. PMID- 17415494 TI - A new anode for solid oxide fuel cells with enhanced OCV under methane operation. AB - A new SOFC anode material based upon oxygen excess perovskite related phases has been synthesised. The material shows better electrochemical performance than other alternative new anodes and comparable performance to the state-of-art of the electrodes, Ni-YSZ cermets, under pure hydrogen. Furthermore, this material shows an enhanced performance under methane operation with high open circuit voltages, i.e. 1.2-1.4 V at 950 degrees C, without using steam excess. The effect of the anode configuration was tested in one and four layer configurations. The optimised electrode polarisation resistances were just 0.12 ohm cm(2) and 0.36 ohm cm(2), at 950 degrees C, in humidified H(2) and humidified CH(4), respectively. Power densities of 0.5 W cm(-2) and 0.35 W cm(-2) were obtained in the same conditions. A very low anodic overpotential of 100 mV at 1 A cm(-2) was obtained under humidified H(2) at 950 degrees C. Samples were tested for two days in reducing and oxidising conditions, alternating heating and cooling processes from 850 degrees C to 950 degrees C, showing stable electrode performance and open circuit voltages. The results show that the substituted strontium titanates are very promising anode materials for SOFC. PMID- 17415495 TI - In situ solid state 11B MAS-NMR studies of the thermal decomposition of ammonia borane: mechanistic studies of the hydrogen release pathways from a solid state hydrogen storage material. AB - The mechanism of hydrogen release from solid state ammonia borane (AB) has been investigated via in situ solid state (11)B and (11)B{(1)H} MAS-NMR techniques in external fields of 7.1 T and 18.8 T at a decomposition temperature of 88 degrees C, well below the reported melting point. The decomposition of AB is well described by an induction, nucleation and growth mechanistic pathway. During the induction period, little hydrogen is released from AB; however, a new species identified as a mobile phase of AB is observed in the (11)B NMR spectra. Subsequent to induction, at reaction times when hydrogen is initially being released, three additional species are observed: the diammoniate of diborane (DADB), [(NH(3))(2)BH(2)](+)[BH(4)](-), and two BH(2)N(2) species believed to be the linear (NH(3)BH(2)NH(2)BH(3)) and cyclic dimer (NH(2)BH(2))(2) of aminoborane. At longer reaction times the sharper features are replaced by broad, structureless peaks of a complex polymeric aminoborane (PAB) containing both BH(2)N(2) and BHN(3) species. The following mechanistic model for the induction, nucleation and growth for AB decomposition leading to formation of hydrogen is proposed: (i) an induction period that yields a mobile phase of AB caused by disruption of the dihydrogen bonds; (ii) nucleation that yields reactive DADB from the mobile AB; and (iii) growth that includes a bimolecular reaction between DADB and AB to release the stored hydrogen. PMID- 17415496 TI - Mesoporous and nanowire Co3O4 as negative electrodes for rechargeable lithium batteries. AB - The conversion reactions associated with mesoporous and nanowire Co(3)O(4) when used as negative electrodes in rechargeable lithium batteries have been investigated. Initially, Li is intercalated into Co(3)O(4) up to x approximately 1.5 Li in Li(x)Co(3)O(4). Thereafter, both materials form a nanocomposite of Co particles imbedded in Li(2)O, which on subsequent charge forms CoO. The capacities on cycling increase on initial cycles to values exceeding the theoretical value for Co(3)O(4) + 8 Li(+) + 8e(-) --> 4 Li(2)O + 3 Co, 890 mAhg( 1), and this is interpreted as due to charge storage in a polymer layer that forms on the high surface area of nanowire and mesoporous Co(3)O(4). After 15 cycles, the capacity decreases drastically for the nanowires due to formation of grains that are separated one from another by a thick polymer layer, leading to electrical isolation. In contrast, the mesoporous Co(3)O(4) losses its mesoporosity and forms a morphology similar to bulk Co(3)O(4) (Co particles imbedded in Li(2)O matrix) with which it exhibits a similar capacity on cycling. In contrast to mesoporous lithium intercalation compounds, which show superior capacity at high rates compared to bulk materials, mesoporosity does not seem to improve the capacity of conversion reactions on extended cycling. If, however, mesoporosity could be retained during the conversion reaction, then higher capacities could be obtained in such systems. PMID- 17415497 TI - Dependence of the photoelectrochemical performance of sensitised ZnO on the crystalline orientation in electrodeposited ZnO thin films. AB - The influence of the crystal orientation in porous crystalline films of ZnO electrodeposited on the photoelectrochemical characteristics of the films is studied. For differently oriented ZnO thin films following removal of the respective structure-directing agent (SDA) and adsorption of a sensitiser, time resolved photocurrent measurements, intensity modulated photocurrent spectroscopy (IMPS), intensity modulated photovoltage spectroscopy (IMVS) and current-voltage curves were measured in acetonitrile-based electrolytes containing I(3)(-)/I(-) as the redox electrolyte. The crystal orientation has a significant influence on the charge transport across such films and hence is reflected in the observed electrode kinetics. Films originally grown in the presence of, e.g., Coumarin 343 as a SDA, showed a significantly faster response to illumination. Increased electron diffusion coefficients and diffusion lengths were calculated from the results of IMPS and IMVS, caused by a faster electron movement in the films. Implications of these findings on further improvements of sensitised ZnO films prepared by electrochemical deposition are discussed. PMID- 17415498 TI - The influence of membrane electrode assembly water content on the performance of a polymer electrolyte membrane fuel cell as investigated by 1H NMR microscopy. AB - The relation between the performance of a self-humidifying H(2)/O(2) polymer electrolyte membrane fuel cell and the amount and distribution of water as observed using (1)H NMR microscopy was investigated. The integrated (1)H NMR image signal intensity (proportional to water content) from the region of the polymer electrolyte membrane between the catalyst layers was found to correlate well with the power output of the fuel cell. Several examples are provided which demonstrate the sensitivity of the (1)H NMR image intensity to the operating conditions of the fuel cell. Changes in the O(2)(g) flow rate cause predictable trends in both the power density and the image intensity. Higher power densities, achieved by decreasing the resistance of the external circuit, were found to increase the water in the PEM. An observed plateau of both the power density and the integrated (1)H NMR image signal intensity from the membrane electrode assembly and subsequent decline of the power density is postulated to result from the accumulation of H(2)O(l) in the gas diffusion layer and cathode flow field. The potential of using (1)H NMR microscopy to obtain the absolute water content of the polymer electrolyte membrane is discussed and several recommendations for future research are provided. PMID- 17415499 TI - Exposure and health risk of gallium, indium, and arsenic from semiconductor manufacturing industry workers. PMID- 17415500 TI - Quantitative structure-activity relationships for predicting the joint toxicity of substituted anilines and phenols to algae. PMID- 17415501 TI - Arsenic distribution in a tilapia (Oreochromis mossambicus) water-sediment aquacultural ecosystem in blackfoot disease hyperendemic areas. PMID- 17415502 TI - Interaction between a synthetic dye bath and selenium in their toxicity to Dunaliella tertiolecta under two light intensities. PMID- 17415504 TI - Comment on van Brummen et al.: The effect of vaginal and cesarean delivery on lower urinary tract symptoms: what makes the difference? PMID- 17415505 TI - Accurate diagnosis of acute abdomen in FMF and acute appendicitis patients: how can we use procalcitonin? AB - This study was conducted to define the value of procalcitonin (PCT) levels in the differential diagnosis of abdominal familial Mediterranean fever (FMF) attacks from acute appendicitis. From October 2006 to January 2007, 28 FMF (12 males, 16 females) patients with acute abdominal attacks and 34 patients (18 males) with acute abdomen who underwent operation with the clinical diagnosis of acute appendicitis were consecutively enrolled in this study. FMF patients with concurrent infectious diseases were excluded. PCT values were measured by an immunofluorescent method using the B.R.A.H.M.S. PCT kit (B.R.A.H.M.S. Diagnostica, Berlin, Germany). Erythrocyte sedimentation rate (ESR), C-reactive proteins (CRP) and leucocyte levels were also noted. Mean disease duration in FMF patients was 9.6 +/- 8.1 years (range 2-33 years) and all were on colchicine therapy with a mean colchicine dosage of 1.2 +/- 0.4 mg/day. Among the operated patients, 5 were excluded: 3 patients had normal findings and 2 had intestinal perforation (PCT levels were 2.69 and 4.93 ng/ml, respectively) at operative and pathologic evaluation. There were no significant differences between the two groups with respect to gender and age (p was not significant (NS) for all). Acute phase reactants and PCT levels were increased in patients with FMF compared to patients with acute appendicitis (0.529[0.12 +/- 0.96] vs 0.095 [0.01-0.80] p < 0.001, respectively). PCT levels higher than 0.5 ng/ml were found in 11% (3/28) of FMF patients compared to 62% (18/29) of acute appendicitis patients (p < 0.001). Our results suggest that PCT could be a useful test in the differentiation of abdominal FMF attacks from acute appendicitis, though it should not supplant more conventional investigations. PMID- 17415506 TI - Accrual of organ damage over time in Argentine patients with systemic lupus erythematosus: a multi-centre study. AB - The goals of this study were to ascertain damage in patients with systemic lupus erythematosus (SLE) from five rheumatologic centres in Argentina and to examine overall damage, damage by domain and damage by item within each domain. We performed a retrospective observational study including patients with SLE (ACR 1997 revised and modified criteria) from five rheumatology centres in Argentina. Organ damage was scored using the SLICC/ACR DI (SDI), ascertained at years 1, 2, 5 and 10. Three centres provided information up to the fifth year. Of the 197 patients, 88.3% were women and their mean age was 33.2 years. The mean disease duration and follow-up were 7.6 and 5.3 years, respectively. Damage accrued gradually over time with SDI ranging from 0.52 (+/-1.1) at year 1 up to 2.46 (+/ 2.1) at year 10. The renal system was the most involved system, followed by the neuropsychiatric, the cardiovascular and the musculoskeletal systems. Proteinuria, cognitive impairment, pericarditis, avascular necrosis, cataract and alopecia were the predominant items in their respective systems. Systems such as peripheral vascular, pulmonary, gastrointestinal, diabetes, malignancy and premature gonadal failure were not frequent. Overall SDI had a gradual increase over time. Damage in each domain of SDI, except for diabetes, had a similar behaviour. Behaviour of items in each domain varied. PMID- 17415507 TI - Microscopic polyangiitis and myasthenia gravis: the battle of Occam and Hickam. AB - Microscopic polyangiitis (MPA) is a pauci-immune, necrotising, small-vessel vasculitis with an incidence of 3.6 per million population that typically presents in adulthood. Myasthenia gravis (MG), the most common disorder of the neuromuscular junction is rare, with an incidence of four per million population. We present the case of an adolescent girl previously diagnosed with MPA at age 7 years who presented with breathlessness and respiratory failure aged 15 years. The respiratory symptoms were due to thymoma-MG, which was successfully treated with cholinesterase inhibitors and thymectomy. This case report illustrates that the well-established doctrines of Occam's razor and of 'common conditions occurring commonly' are not universally applicable, and that in the adolescent age group, one should still consider Hickam's dictum. PMID- 17415508 TI - Myelo-optico-neuropathy in copper deficiency occurring after partial gastrectomy. Do small bowel bacterial overgrowth syndrome and occult zinc ingestion tip the balance? AB - Acquired copper deficiency has recently been recognized as a cause of myeloneuropathy mimicking subacute combined degeneration due to vitamin B-12 deficiency. A remote history of gastric surgery is frequently associated with this syndrome. However, the very limited prevalence of severe copper deficiency in patients with a history of gastric surgery suggests that additional contributing factors are likely to be involved. We describe a patient with copper deficiency and a previous Billroth II partial gastrectomy for gastric carcinoma, presenting with severe myelo-optico-neuropathy, demyelinating lesions of the brain, and subjective hyposmia. An abnormal glucose breath test also revealed small bowel bacterial overgrowth syndrome. Copper replacement therapy associated with antibiotic therapy was effective in preventing further neurological damage and in obtaining mild improvement. We propose that copper status should be evaluated in all patients presenting with unexplained noninflammatory myeloneuropathy. Small bowel bacterial overgrowth syndrome should be investigated as a cause of generalized malabsorption and a possible contributing factor to copper deficiency after gastric surgery, as should occult zinc ingestion. PMID- 17415509 TI - N-acetylaspartic acid in cerebrospinal fluid of multiple sclerosis patients determined by gas-chromatography-mass spectrometry. AB - BACKGROUND: Axonal degeneration is considered to play a major role in the development of clinical disability in multiple sclerosis (MS). N-AcetylAspartic Acid (NAA) is a neuron-specific marker constantly identified in MR-spectroscopy studies of the normal and MS brain. To our knowledge there are no studies available that evaluated NAA in cerebrospinal fluid (CSF) as a possible marker for disease severity. OBJECTIVE: To evaluate CSF concentrations of NAA in MS in relation to disease phenotype, clinical measures of disability and MRI markers of disease burden. METHODS: NAA concentrations were determined in CSF of 46 patients with MS (26 relapsing remitting (RRMS), 12 secondary progressive (SPMS) and 8 primary progressive (PPMS)). Prior to lumbar puncture, MS-patients underwent MRI and clinical examination, including the Expanded Disability Status Scale (EDSS) and the MS Functional Composite (MSFC). Additionally, CSF concentrations of NAA were determined in 12 patients with other neurological diseases (OND). RESULTS: Median CSF NAA concentration was 0.74 (IQR: 0.59-0.94) in RRMS , 0.54 (IQR: 0.35 0.73) in SPMS and 0.83 mumol/l (IQR: 0.56-1.03) in PPMS patients. SPMS patients had a significantly lower NAA concentration than RRMS patients. NAA concentrations correlated with EDSS (r = )0.37, p = 0.016), MSFC (r = 0.41, p = 0.010), normalised brain volume (r = 0.49, p = 0.001), T2 lesion load (r = )0.35, p = 0.021) and black hole lesion load (r = )0.47, p = 0.002). No differences were observed between OND (median: 0.57 IQR: 0.28-0.73) and MS patients. CONCLUSION: CSF NAA concentration in MS patients is related to clinical performance and MRI measures of disease burden and may therefore be an important neuron specific marker of disease severity and possibly progression. PMID- 17415510 TI - A novel mutation at the N-terminal of SMN Tudor domain inhibits its interaction with target proteins. AB - Although most patients with spinal muscular atrophy (SMA) are homozygous for deletion of the SMN1 gene, some patients bear one SMN1 copy with a subtle mutation. Detection of such an intragenic mutation may be helpful not only in confirming diagnosis but also in elucidating functional domains of the SMN protein. In this study, we identified a novel mutation in SMN1 of two Japanese patients with type I SMA. DHPLC and sequencing analysis revealed that they harbored a point mutation in SMN1 exon 3, 275G > C, leading to tryptophan-to serine substitution at amino acid 92 (W92S) at the Nterminal of SMN Tudor domain. In-vitro protein binding assays showed that the mutation severely reduced interaction of the domain with SmB protein and fibrillarin, suggesting that it impairs the critical function of SMN. In conclusion, we reported here that a novel mutation, W92S, in the Tudor domain affects the interaction of SMN with the target proteins. PMID- 17415511 TI - Transcranial ultrasound in different monogenetic subtypes of Parkinson's disease. AB - Hyperechogenicity of the substantia nigra (SN) has been found to be a typical sign in idiopathic Parkinson's disease (PD), prevalent in more than 90% of affected individuals. To see whether SN hyperechogenicity is also characteristic for monogenetically caused PD, we investigated PD patients with alpha-synuclein, LRRK2, parkin, PINK1 and DJ-1 mutations by transcranial sonography (TCS). In all these patients the area of SN echogenicity was significantly larger than in healthy controls, but smaller, than in idiopathic PD. As SN hyperechogenicity could be related to an increased iron content of the SN, these findings suggest that iron may play a less significant role in the pathogenesis of monogenetically caused compared to idiopathic PD. PMID- 17415512 TI - Headaches during angiography and endovascular procedures. AB - BACKGROUND: The occurrence of headaches during cerebral angiography or endovascular procedures is an ill-defined issue, as limited information is available to define its frequency, risk factors, pathogenesis and implications for future pain management. PURPOSE: To determine the frequency of headaches during endovascular procedures (HdEVP) and to define their characteristics and risk factors for their occurrence. METHODS: Prospective cross-sectional observational study of adult patients undertaking cerebral angiography or endovascular procedures, recording HdEVP clinical features, previous headache history and procedural variables. RESULTS: One hundred and twenty two procedures were evaluated. HdEVP occurred in 13 patients, both in diagnostic and therapeutic procedures. Pain was described as brief, stabbing or localized pressure of mild to moderate intensity, felt ipsilaterally to the manipulated vessel. Its occurrence was associated with therapeutic interventions (p = 0.007), female gender (p = 0.015) and previous history of more than 4 headache episodes per month (p = 0.018). CONCLUSION: HdEVP is an uncommon brief headache that should be further evaluated in the future. Its pathogenesis is probably related to mechanical vessel distension, which activates the trigeminovascular nociceptive system in susceptible individuals. PMID- 17415514 TI - Dielectric relaxation spectrum of poly (epsilon-caprolactone) networks hydrophilized by copolymerization with 2-hydroxyethyl acrylate. AB - The dielectric relaxation spectrum of polycaprolactone (PCL) networks hydrophilized with different amounts of 2-hydroxyethyl acrylate (HEA) is investigated. PCL is a semicrystalline polyester with a complex relaxation spectrum that includes the main alpha relaxation and two secondary modes (beta, gamma) at lower temperatures. The overlapping of the different relaxational modes was split by using several Havriliak-Negami functions. Crosslinking the material modifies the dynamics of the main relaxation process as reflected by the parameters that characterize the Vogel behavior of the process and the dynamic fragility. The incorporation of HEA units in the network results in a material with microphase separation: two alpha processes are detected, the one corresponding to the PCL chains and the new one associated to nanometric regions that contain different amount of both comonomers. The incorporation of the HEA units in the system involves the presence of a new beta(sw) relaxation due to the link of two side chains by water molecules through hydrogen bonding. PMID- 17415516 TI - Predicting siRNA efficiency. AB - Since the identification of RNA-mediated interference (RNAi) in 1998, RNAi has become an effective tool to inhibit gene expression. The inhibition mechanism is triggered by introducing a short interference double-stranded RNA (siRNA,19 approximately 27 bp) into the cytoplasm, where the guide strand of siRNA (usually antisense strand) binds to its target messenger RNA and the expression of the target gene is blocked. RNAi has been widely applied in gene functional analysis, and as a potential therapeutic strategy in viral diseases, drug target discovery, and cancer therapy. Among the factors which may compromise inhibition efficiency, how to design siRNAs with high efficiency and high specificity to its target gene is critical. Although many algorithms have been developed for this purpose, it is still difficult to design such siRNAs. In this review, we will briefly discuss prediction methods for siRNA efficiency and the problems of present approaches. PMID- 17415517 TI - The neural basis of functional neuroimaging signal with positron and single photon emission tomography. AB - Functional imaging techniques such as positron and single-photon emission tomography exploit the relationship between neural activity, energy demand and cerebral blood flow to functionally map the brain. Despite the fact that neurobiological processes are not completely understood, several results have revealed the signals that trigger the metabolic and vascular changes accompanying variations in neural activity. Advances in this field have demonstrated that release of the major excitatory neurotransmitter glutamate initiates diverse signaling processes between neurons, astrocytes and blood perfusion, and that this signaling is crucial for the occurrence of brain imaging signals. Better understanding of the neural sites of energy consumption and the temporal correlation between energy demand, energy consumption and associated cerebrovascular hemodynamics gives novel insight into the potential of these imaging tools in the study of metabolic neurodegenerative disorders. PMID- 17415518 TI - A systems biology approach to genetic studies of pancreatitis and other complex diseases. AB - Pancreatitis is usually inflammation of the pancreas without infection. Our understanding of pancreatitis has been built on autopsy studies, surgical biopsies and surrogate markers of inflammation and fibroses, including abdominal imaging techniques and pancreatic functional studies. However, the discovery that a number of different environmental factors and various genetic abnormalities are seen in patients with similar appearing pancreatitis phenotypes teaches us that end-stage pathology is not the disorder. Understanding complex associations and interactions requires that the components and their interactions be organized, stratified and functionally defined. Systems biology, in the broad sense, provides the approach and tools to define the complex mechanisms driving pathology. As the mathematics behind these pathways and mechanisms are defined and calibrated, the potential pathology of patients with early signs of disease can be predicted, and a number of patient-specific targets for intervention can be defined. PMID- 17415519 TI - Cancer systems biology: exploring cancer-associated genes on cellular networks. AB - Genomic alterations lead to cancer complexity and form a major hurdle for comprehensive understanding of the molecular mechanisms underlying oncogenesis. In this review, we describe recent advances in studying cancer-associated genes from a systems biology point of view. The integration of known cancer genes onto protein and signaling networks reveals the characteristics of cancer genes within networks. This approach shows that cancer genes often function as network hub proteins which are involved in many cellular processes and form focal nodes in information exchange between many signaling pathways. Literature mining allows constructing gene-gene networks, in which new cancer genes can be identified. The gene expression profiles of cancer cells are used for reconstructing gene regulatory networks. By doing so, genes which are involved in the regulation of cancer progression can be picked up from these networks, after which their functions can be further confirmed in the laboratory. PMID- 17415520 TI - Network analysis approach for biology. AB - The biological system is a complex physicochemical system consisting of numerous dynamic networks of biochemical reactions and signaling interactions between cellular components. This complexity makes it virtually unanalyzable by traditional methods. Hence, biological networks have been developed as a platform for integrating information from high- to low-throughput experiments for analysis of biological systems. The network analysis approach is vital for successful quantitative modeling of biological systems. The numerous online pathway databases vary widely in coverage and representation of biological processes. An integrated network-based information system for querying, visualization and analysis promised successful integration of data on a large scale. Such integrated systems will greatly facilitate the understanding of biological interactions and experimental verification. PMID- 17415522 TI - Hepatocyte growth factor in invasive growth of carcinomas. AB - Hepatocyte growth factor is a multifunctional cytokine of the tumor microenvironment. An important advance in the knowledge of cancer progression has been the appreciation that the tumor invasive phenotype is strongly influenced by microenvironmental stimuli. Malignant tumor cells recruit vasculature and stroma through the production of growth factors and cytokines. The locally activated microenvironment (both cellular and extracellular elements) in turn modifies the proliferative and invasive behavior of the tumor cells. Hepatocyte growth factor accomplishes most of the functions of the invasive program in carcinomas (loss of adhesive junctions, motility, angiogenesis, survival/apoptosis), and may interact with other signals such as hypoxia. The purpose of the present review is to highlight examples of the progress in this area. The influence of hepatocyte growth factors on the carcinoma invasive phenotype is considered by evaluating the gene targets and the network of transcription factors activated in the specific responses. PMID- 17415523 TI - Monothiol glutaredoxins: a common domain for multiple functions. AB - Monothiol glutaredoxins with the CGFS sequence at the active site are widespread among prokaryotes and eukaryotes. Two subclasses exist, those with a single glutaredoxin domain and those with a thioredoxin-like region followed by one or more glutaredoxin domains. Studies in Saccharomyces cerevisiae have demonstrated the role of the Grx5 protein in the biogenesis of iron-sulfur clusters. Grx5 homologues in other eukaryotes could carry out similar functions. Two S. cerevisiae monothiol glutaredoxins with the thioredoxin-like extension, Grx3 and Grx4, are modulators of the transcriptional activator Aft1, which regulates iron uptake in yeast. The human PICOT protein is a Grx3/Grx4 homologue with the same hybrid primary structure that regulates protein kinase C activity and may participate in physiological processes such as control of cardiac function. Therefore, monothiol glutaredoxins share a common basic structural motif and biochemical mechanism of action, while participating in a diversity of cellular functions as protein redox regulators. PMID- 17415524 TI - FABP7 expression in glioblastomas: relation to prognosis, invasion and EGFR status. AB - FABP7 expression has been analysed in a series of 123 glioblastomas (68 pure GBM, 55 GBMO, i.e. with oligodendroglial component). FABP7, found in 91/123 samples, showed a pure cytoplasmic expression in 69 cases, and cytoplasmic+nuclear expression in 22 cases. FABP7 expression was associated with pure GBM histology and shorter survival (15.7 months versus 21.5 months). Nuclear expression of FABP7 was more specifically related to EGFR amplification and more invasive tumors. These data, although they need to be confirmed by further studies, support the relation between FABP7, astrocytic features, invasion and poor prognosis and suggests that EGFR amplification is associated with nuclear translocation of FABP7. PMID- 17415525 TI - The anti-cancer compound Nordy inhibits CXCR4-mediated production of IL-8 and VEGF by malignant human glioma cells. AB - The chemokine receptor CXCR4 plays an important role in tumor growth, angiogenesis and metastasis. Our previous studies showed that Nordy, a synthetic chiral compound of nordihydroguaiaretic acid, inhibited the growth and angiogenesis of various malignant tumors. In this study we examined the capacity of Nordy to regulate CXCR4-mediated production of angiogenic factors by human glioblastoma cells. We found that Nordy potently inhibited CXCR4 ligand SDF-1 induced production of IL-8 and vascular endothelial cell growth factor, two important angiogenic factors implicated in the progression of malignant tumors. Further study revealed that the effect of Nordy was attributable to its down regulation of the expression of functional CXCR4 in glioblastoma cells. These results suggest that the anti-cancer activity of Nordy is due, at least in part, to its suppression of the chemokine receptor CXCR4 thus reducing the production of angiogenic factors by tumor cells. PMID- 17415526 TI - Regulation of tumor pH and the role of carbonic anhydrase 9. AB - The high metabolic rate required for tumor growth often leads to hypoxia in poorly-perfused regions. Hypoxia activates a complex gene expression program, mediated by hypoxia inducible factor 1 (HIF1alpha). One of the consequences of HIF1alpha activation is up-regulation of glycolysis and hence the production of lactic acid. In addition to the lactic acid-output, intracellular titration of acid with bicarbonate and the engagement of the pentose phosphate shunt release CO(2) from cells. Expression of the enzyme carbonic anhydrase 9 on the tumor cell surface catalyses the extracellular trapping of acid by hydrating cell-generated CO(2) into [see text] and H(+). These mechanisms contribute towards an acidic extracellular milieu favoring tumor growth, invasion and development. The lactic acid released by tumor cells is further metabolized by the tumor stroma. Low extracellular pH may adversely affect the intracellular milieu, possibly triggering apoptosis. Therefore, primary and secondary active transporters operate in the tumor cell membrane to protect the cytosol from acidosis. We review mechanisms regulating tumor intracellular and extracellular pH, with a focus on carbonic anhydrase 9. We also review recent evidence that may suggest a role for CA9 in coordinating pH(i) among cells of large, unvascularized cell clusters. PMID- 17415527 TI - Regulation of DNA repair in hypoxic cancer cells. AB - Emerging evidence indicates that the tumor microenvironmental stress of hypoxia can induce genetic instability in cancer cells. We and others have found that the expression levels of key genes within the DNA mismatch repair (MMR) and homologous recombination (HR) pathways are coordinately repressed by hypoxia. These decreases are associated with functional impairments in both MMR and HR repair under hypoxic conditions, and thus they represent a possible mechanistic explanation for the observed phenomenon of hypoxia-induced genetic instability. In parallel, studies also indicate that several DNA damage response factors are activated in response to hypoxia and subsequent reoxygenation, including ATM/ATR, Chkl/Chk2 and BRCA1. Taken together, these findings reveal that hypoxia induces a unique cellular stress response involving an initial, acute DNA damage response to hypoxia and reoxygenation, followed by a chronic response to prolonged hypoxia in which selected DNA repair pathways are coordinately suppressed. In this review, we discuss these pathways and the possible mechanisms involved, as well as the consequences for genetic instability and tumor progression within the tumor microenvironment. PMID- 17415528 TI - Effects of hypoxia on tumor metabolism. AB - Rapidly growing tumors invariably contain hypoxic regions. Adaptive response to hypoxia through angiogenesis, enhanced glucose metabolism and diminished but optimized mitochondrial respiration confers survival and growth advantage to hypoxic tumor cells. In this review, the roles of hypoxia, the hypoxia inducible factors, oncogenes and tumor suppressors in metabolic adaptation of tumors are discussed. These new insights into hypoxic metabolic alterations in tumors will hopefully lead us to target tumor bioenergetics for the treatment of cancers. PMID- 17415529 TI - Targeting hypoxia cell signaling for cancer therapy. AB - Hypoxia, a decrease in oxygen levels, is a hallmark of solid tumors. Hypoxic cells are more resistant to killing by ionizing radiation and chemotherapy, are more invasive and metastatic, resistant to apoptosis, and genetically unstable. Over the last two decades, the discovery of Hypoxia Inducible Factors, a family of transcription factors crucially involved in the response of mammalian cells to oxygen deprivation, has led to the identification of a molecular target associated with hypoxia suitable for the development of cancer therapeutics. These features of solid tumors may offer a unique opportunity for selective therapeutic approaches. A number of strategies targeting hypoxia and/or Hypoxia Inducible Factors (HIF) have been developed over the last several years and will be described. The exponentially growing interest in therapeutic strategies targeting hypoxia/HIF will undoubtedly generate more active compounds for preclinical and clinical development. A rational development plan aimed to validate target inhibition in preclinical models and early clinical trials is essential for a rapid translation of these agents to the treatment of human cancers. PMID- 17415530 TI - Retinal pigment epithelial tear after intravitreal ranibizumab for subfoveal CNV secondary to AMD. AB - PURPOSE: To report a case of retinal pigment epithelial tear following intravitreal ranibizumab injection for subfoveal choroidal neovascularization. METHODS: Retrospective single case review. RESULTS: A 78-year-old Caucasian female was treated with intravitreal ranibizumab for occult subfoveal choroidal neovascularization (CNV) from age-related macular degeneration (AMD). She returned for evaluation with decreased vision and was found to have a retinal pigment epithelial tear on biomicroscopy. Fluorescein angiography and OCT testing confirmed the clinical findings. CONCLUSION: Although a pigment epithelial tear in neovascular AMD can represent natural history, prior reports of such tears after thermal laser, photodynamic therapy with verteporfin and following intravitreal injection of pegaptanib Na combined with this case report suggest that clinicians should be aware of and monitor patients for the possibility of this complication after intravitreal injections of ranibizumab. PMID- 17415531 TI - Late dislocation of an anterior chamber intraocular lens into the vitreous. AB - We report the case of a patient with dislocation of an anterior chamber intraocular lens (AC IOL) into the vitreous, 5 years after surgery. The cause of the dislocation may have been the size of the pupil and habitual eye rubbing. This case report describes a rare complication of AC IOL implantation in eyes of previous iris lesions or pupil abnormalities. This complication should be kept in mind when an AC IOL implantation is planned for the correction of aphakia. PMID- 17415532 TI - Development of a large scale process for the conversion of polysialogangliosides to monosialotetrahexosylganglioside with a novel strain of Brevibacterium casei producing sialidase. AB - A bioconversion process of producing GM1 (monosialotetrahexosylganglioside) on an industrial scale was developed with a novel sialidase-producing strain Brevibacterium casei. The sialidase hydrolyzed polysialogangliosides to produce GM1 but did not act on GM1. When Brevibacterium casei was cultured in a synthetic medium containing crude pig brain gangliosides (10% w/v) at 30 degrees C for 24 h in a 50 l fermenter, most of the polysialogangliosides were converted to GM1. The content of GM1 was increased from 9% in crude gangliosides to 45% with 70% (w/w) yield. PMID- 17415533 TI - The neuroprotective effect of Withania somnifera root extract in MPTP-intoxicated mice: an analysis of behavioral and biochemical variables. AB - We studied the influence of Withania somnifera (Ws) root extract (100 mg/kg body weight) on parkinsonism induced by 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP; i.p, 20 mg/kg body weight for 4 days), via the analysis of behavioral features and the oxidant-antioxidant imbalance in the midbrain of mice. A significant alteration in behavior, increased levels of thiobarbituric acid reactive substance (TBARS), and increased activities of superoxide dismutase (SOD) and catalase (CAT) were noticed in this region of brain in MPTP-treated mice. Oral treatment with the root extract resulted in a significant improvement in the mice's behavior and antioxidant status, along with a significant reduction in the level of lipid peroxidation. The results indicated that at least part of the chronic stress-induced pathology may be due to oxidative stress, which is mitigated by Ws. Further studies are needed to assess the precise mechanism to support the clinical use of the plant as an antiparkinsonic drug. PMID- 17415534 TI - The construction of the eukaryotic expression plasmid pcDNA3.1/azurin and the increased apoptosis of U2OS cells transfected with it. AB - In our previous study, we demonstrated that azurin could selectively trigger apoptosis in human osteosarcoma cell line U2OS cells. However, the rate of apoptosis (35.8 +/- 3.2%) is not very high, and azurin is too expensive to obtain readily. To solve these problems, we constructed a eukaryotic expression plasmid containing the azurin gene with an influenza virus haemagglutinin 9 peptide HA epitope tag, and transfected the recombinant plasmid pcDNA3.1(+)/azurin into U2OS cells. RT-PCR and Western blot analysis validated the successful transfection and the expression of the azurin-HA protein. Conspicuous apoptosis of the transfected cells was detected by flow cytometry (FCM) and the DNA ladder test. The apoptosis rate reached 64.3 +/- 13.1%. The transcriptional levels of the Bax and p53 genes increased significantly in U2OS cells transfected with pcDNA3.1(+)/azurin, but the Bcl-2 mRNA level decreased. There was no difference in the levels of Bcl-xl mRNA and Survivin mRNA. We propose that the transfection of the recombinant plasmid pcDNA3.1(+)/azurin can significantly induce apoptosis in U2OS cells. This is closely associated with the up-regulation of the transcriptional level of the Bax and p53 genes, and the down-regulation of that of the Bcl-2 gene. PMID- 17415535 TI - [Condylar resection alone versus alloplastic reconstruction. Comparison of long term functional and cosmetic results]. AB - After condylar resection a decision on whether to proceed immediately to reconstruction has to be decided. In this paper, two patients who had undergone hemimandibulectomy including exarticulation, in one because of an expanded keratocystic odontogenic tumour and in the other because of oral squamous cell carcinoma, are presented. In one patient a metallic condylar reconstruction plate combined with an iliac crest graft was implanted for primary mandibular reconstruction, whereas in the other the part of the mandible that had been removed and the condylar head were not replaced. One patient was followed up for 5 years and the other for 6 years. Functional (max. incisal distance, protrusive and lateral excursions, occlusion and joint noises) and cosmetic results (scarring, facial nerve function), and also quality of life with and without primary mandibular replacement by a metallic condylar reconstruction plate are compared. PMID- 17415536 TI - [Attention deficit and hyperactivity disorder: what is currently known and its significance for ENT specialists]. AB - Attention deficit hyperactivity disorder (ADHD) is a behavioural disorder characterised by marked problems with inattention and increased impulsiveness, with or without hyperactivity; in most cases it becomes obvious in nursery-school age children, and the latest point for its initial appearance is while the child affected is at primary school. Even in children of normal intelligence, the cognitive, emotional, and social development is decidedly limited, which can have corresponding ramifications for adult life. If left untreated it can progress to dissocial development and give the impression of an impulsive personality disorder in adulthood.ENT specialists are also confronted with children, adolescents and even adults in whom ADHD has already been diagnosed in the course of investigations/treatment. If the suspicion of hearing impairment as the cause of problems in daily life is not confirmed in a patient not hitherto known to have ADHD ENT specialists should also consider ADHD in the differential diagnosis. In this report the diagnostic and therapeutic strategies currently applied for ADHD and their importance for ENT specialists are presented. PMID- 17415537 TI - [Case allocation of extensive operations on head and neck within the German DRG system 2004-2007: what is the net result of the continued developments in case allocation?]. AB - BACKGROUND: When the German DRG system was implemented there was some doubt about whether patients with extensive head and neck surgery would be properly accounted for. Significant efforts have therefore been invested in analysis and case allocation of those in this group. The object of this study was to investigate whether the changes within the German DRG system have led to improved case allocation. METHODS: Cost data received from 25 ENT departments on 518 prospective documented cases of extensive head and neck surgery were compared with data from the German institute dealing with remuneration in hospitals (InEK). Statistical measures used by InEK were used to analyse the quality of the overall system and the homogeneity of the individual case groups. RESULTS: The reduction of variance of inlier costs improved by about 107.3% from the 2004 version to the 2007 version of the German DRG system. The average coefficient of cost homogeneity rose by about 9.7% in the same period. Case mix index and DRG revenues were redistributed from less extensive to the more complex operations. Hospitals with large numbers of extensive operations and university hospitals will gain most benefit from this development. CONCLUSION: Appropriate case allocation of extensive operations on the head and neck has been improved by the continued development of the German DRG system culminating in the 2007 version. Further adjustments will be needed in the future. PMID- 17415538 TI - [Propionic acidemia and sensorineural hearing loss: is there a connection at the molecular genetics level?]. AB - CURRENT KNOWLEDGE: Propionic acidemia is caused by a gene defect leading to malfunction of the enzyme propionyl-CoA carboxylase (PCC) and in turn to a pathologic accumulation of propionic acid. Many mutations have been found at the molecular genetic level over the past 20 years, and their implications for the limitation of enzyme activity of PCC in propionic acidemia are discussed. SCIENTIFIC QUESTION AND AIMS OF THE STUDY: As an elevated incidence of deafness has been observed in patients with propionic acidemia, the question arises of whether mutations primarily responsible for this disease could also be the underlying cause for a genetic form of deafness. METHODS AND RESULTS: As well as a standard pure tone audiogram, a pedigree was elaborated and DNA isolated for each family concerned. In one family several subjects displayed mutations of both the PCCA and the PCCB -subunits; these included only one girl whose phenotype was affected, however. CONCLUSIONS: Mutation of the PCCB subunit p.R113X has not previously been mentioned in the literature. According to our present knowledge no connection can be assumed between either of the two mutations and the severe sensorineural hearing loss. PMID- 17415539 TI - [Swelling of the neck region]. PMID- 17415540 TI - [Implementation of the speech comprehension test for complex syntactic structures (after D.V. Bishop) in 6- and 8-year-old primary school children]. AB - Standardized tests for speech comprehension have been lacking for the diagnostic investigation of primary school children with problems in hearing and speech development. The informal speech comprehension test for complex syntactic structures ("Sprachverstandnistest fur komplexe syntaktische Strukturen", SKSS), a shortened and in Swiss German translated modification of the Test for Reception of Grammar of Bishop, was analyzed for its logical validity of content, difficulty indices, selectivity, reliability and standard values in percent. We conducted the SKSS on 64 primary school children (first grade, average age 6.9 years) and 84 primary school children (second grade, average age 8.2 years). Four test sentences were excluded because of their ambiguous nature. The remaining 26 test sentences most often indicated low difficulty. However, there was an obvious difference between the two groups in terms of difficulty as well reliability. Based on our results, the test proved to be useful for screening. It allows both quantitative and qualitative interpretations. PMID- 17415541 TI - [Form stability after ear correction using the anterior scoring technique: a prospective study to compare anterior and the posterior approaches]. AB - BACKGROUND: Protruding ears are usually caused by an overdeveloped concha and the lack of an anthelical fold. To reconstruct this fold, we applied the open anterior cartilage scoring technique. There are two different approaches for this technique, anterior access or posterior access. We have used the latter for years. After finding some minor partial relapses, we started a prospective study to compare the objective results of both techniques. PATIENTS AND METHOD: Forty patients underwent surgery (20 with anterior access and 20 with posterior access). The position of the ear to the head, which was defined by several standardized points of the ear (distance between the helical rim and the mastoid), was measured preoperatively, 3 weeks postoperatively after removing the anchoring splint sutures, and 3 months postoperatively. RESULTS: The differences between the measurements in the third week and third month show postoperative stability. CONCLUSIONS: We could achieve the same stability using both techniques, but obtained more symmetrical results with posterior access. PMID- 17415542 TI - Molecular defense mechanisms of Barrett's metaplasia estimated by an integrative genomics. AB - Barrett's esophagus is characterized by the replacement of squamous epithelium with specialized intestinal metaplastic mucosa. The exact mechanisms of initiation and development of Barrett's metaplasia remain unknown, but a hypothesis of "successful adaptation" against noxious reflux components has been proposed. To search for the repertoire of adaptation mechanisms of Barrett's metaplasia, we employed high-throughput functional genomic and proteomic methods that defined the molecular background of metaplastic mucosa resistance to reflux. Transcriptional profiling was established for 23 pairs of esophageal squamous epithelium and Barrett's metaplasia tissue samples using Affymetrix U133A 2.0 GeneChips and validated by quantitative real-time polymerase chain reaction. Differences in protein composition were assessed by electrophoretic and mass spectrometry-based methods. Among 2,822 genes differentially expressed between Barrett's metaplasia and squamous epithelium, we observed significantly overexpressed metaplastic mucosa genes that encode cytokines and growth factors, constituents of extracellular matrix, basement membrane and tight junctions, and proteins involved in prostaglandin and phosphoinositol metabolism, nitric oxide production, and bioenergetics. Their expression likely reflects defense and repair responses of metaplastic mucosa, whereas overexpression of genes encoding heat shock proteins and several protein kinases in squamous epithelium may reflect lower resistance of normal esophageal epithelium than Barrett's metaplasia to reflux components. Despite the methodological and interpretative difficulties in data analyses discussed in this paper, our studies confirm that Barrett's metaplasia may be regarded as a specific microevolution allowing for accumulation of mucosal morphological and physiological changes that better protect against reflux injury. PMID- 17415543 TI - [Aspects of pain therapy: a survey of neurologic care in Germany]. AB - BACKGROUND: The state of organized care of neurologic pain patients in Germany was to be established with the the help of our survey. METHODS: A five-page questionnaire was sent to 391 neurologic department heads. RESULTS: Nearly 60% of the departments participated in the survey. Almost all neurologic departments admit pain patients, especially for further diagnostics and neurologic treatment. These are accepted as important neurologic tasks. The quality of care is excellent. For the diagnostics of chronic pain, multidimensional questionnaires are used in nearly 64% of the departments and psychometric tests in almost 70%. Sixty percent of all units document the acute course of pain treatment. Follow-up examinations are performed in 26% of all hospitals. CONCLUSION: Structures exist concerning diagnostics and therapy for neurological in- and outpatients with pain. Care procedures for inpatients with pain need improvement. Outcome is seldom documented. PMID- 17415544 TI - Perturbation of hyaluronan metabolism predisposes patients with type 1 diabetes mellitus to atherosclerosis. AB - AIMS/HYPOTHESIS: Cardiovascular disease contributes to mortality in type 1 diabetes mellitus, but the specific pathophysiological mechanisms remain to be established. We recently showed that the endothelial glycocalyx, a protective layer of proteoglycans covering the endothelium, is severely perturbed in type 1 diabetes, with concomitantly increased plasma levels of hyaluronan and hyaluronidase. In the present study, we evaluated the relationship between hyaluronan and hyaluronidase with carotid intima-media thickness (cIMT), an established surrogate marker for cardiovascular disease. SUBJECTS AND METHODS: Non-smoking type 1 diabetes patients without micro- or macrovascular complications and matched controls were recruited and cIMT of both carotid arteries was measured. To evaluate the relationship between cIMT and hyaluronan and hyaluronidase as well as other parameters, uni- or multivariate regression analyses were performed. RESULTS: We included 99 type 1 diabetes patients (age 10 72 years) and 99 age- and sex-matched controls. Mean cIMT, HbA(1c), high sensitivity C-reactive protein, hyaluronan and hyaluronidase were significantly increased in type 1 diabetes vs controls. Plasma hyaluronan and hyaluronidase were correlated in type 1 diabetes. In univariate regression analyses, mean IMT was associated with plasma hyaluronan, age and male sex, whereas after multivariate analysis only age and sex remained statistically significant. CONCLUSIONS/INTERPRETATION: We conclude that type 1 diabetes patients show structural changes of the arterial wall associated with increased hyaluronan metabolism. These data may lend further support to altered glycosaminoglycan metabolism in type 1 diabetes as a potential mechanism involved in accelerated atherogenesis. PMID- 17415545 TI - The Pro12Ala polymorphism in PPARG2 increases the effectiveness of primary prevention of cardiovascular disease by a lifestyle intervention. PMID- 17415547 TI - Plasminogen activator inhibitor-1 production is pathogenetic in experimental murine diabetic renal disease. AB - AIMS/HYPOTHESIS: Plasminogen activator inhibitor-1 (PAI-1, also known as serpin peptidase inhibitor, clade E [nexin, plasminogen activator inhibitor type 1], member 1 [SERPINE1]) plays a pathogenetic role in renal fibrosis. It is upregulated in experimental and human diabetic nephropathy. These studies assessed the effect of PAI-1 deficiency and overproduction on renal disease in experimental diabetes. MATERIALS AND METHODS: Diabetes was induced by injection of streptozotocin in 6-week-old PAI-1-deficient mice, transgenic mice overexpressing Pai-1 and control mice. Animals were killed after 24 weeks of diabetes or after observation alone. RESULTS: Pai-1 mRNA was upregulated in kidneys from genetically normal mice with diabetes and in non-diabetic Pai-1 transgenic mice. PAI-1 was not further increased in kidneys from Pai-1 transgenic mice with diabetes. Diabetes-associated albuminuria and glomerular injury, as well as renal alpha-smooth muscle actin production, were ameliorated in diabetic PAI-1-deficient mice, an amelioration associated with attenuated increases in renal matrix metallopeptidase-2 expression and activity. Diabetic Pai-1 transgenic mice did not develop increased albuminuria or glomerular injury, but the tubulointerstitial area was modestly enhanced. In addition to the findings in diabetic mice, abnormalities also developed in 30-week-old PAI-1-deficient and Pai-1 transgenic mice without diabetes. PAI-1 deficiency resulted in increased tubulointerstitial area, TGFB1 protein and alpha-smooth muscle actin. Non diabetic 30-week-old Pai-1 transgenic mice developed similar renal abnormalities and increased matrix metallopeptidase-2 activity, together with a modest increase in serum glucose and HbA(1c). CONCLUSIONS/INTERPRETATION: These results demonstrate that endogenous PAI-1 deficiency protects mice from glomerular injury in longer term diabetes and that endogenous PAI-1 maintains normal renal interstitial structure in ageing not associated with diabetes. PMID- 17415548 TI - Glucokinase activator PSN-GK1 displays enhanced antihyperglycaemic and insulinotropic actions. AB - AIMS/HYPOTHESIS: We evaluated the insulinotropic and antihyperglycaemic actions of glucokinase activators (GKAs), especially through acute and subchronic studies in rodent diabetes models with (2R)-2-(4-cyclopropanesulphonylphenyl)-N-(5 fluorothiazol-2-yl)-3-(tetrahydropyran-4-yl)propionamide (PSN-GK1), a novel and potent GKA. MATERIALS AND METHODS: The action of PSN-GK1 on or in the following were investigated: (1) on human liver glucokinase, insulin secretion from MIN6 cells and 2-deoxy-D: -[(3)H]glucose (2-DG) uptake into rat hepatocytes; and (2) in Zucker diabetic fatty rats and in non-diabetic C57Bl/6, diabetic db/db and ob/ob mice. RESULTS: At 5 mmol/l glucose, PSN-GK1 activated glucokinase (4.3 fold, median effective concentration [EC(50)] 130 nmol/l), increased MIN6 insulin secretion (26-fold, EC(50) 267 nmol/l) and 2-DG hepatocytic uptake (threefold, EC(50) 1 micromol/l); at higher glucose concentrations, EC(50)s and fold effectiveness were both lower. In C57Bl/6 mice, PSN-GK1 reduced blood glucose at 1 and 10 mg/kg (by mouth), but insulin was increased significantly at only the higher dose. In hyperinsulinaemic 10-mmol/l glucose clamps, PSN-GK1 increased 2 DG incorporation into liver glycogen sixfold, directly demonstrating liver effects. PSN-GK1 improved glycaemic profiles in db/db mice and Zucker diabetic fatty rats, diabetic animal models in which GKA efficacy has not previously been described, without causing hypoglycaemia. In ob/ob mice, it dose-dependently reduced excursions in OGTTs. Moreover, after subchronic administration, no tachyphylaxis was evident and glycaemia was improved without alterations to lipid levels, liver weight, glycogen content or body weight. CONCLUSIONS/INTERPRETATION: PSN-GK1 was potently antihyperglycaemic through its effects on insulin release and hepatic glucose metabolism. It is one of the most potent GKAs described in the literature and is active in diabetic animal models where GKAs have not been reported to show efficacy to date. Ongoing human trials are investigating the potential of this novel therapeutic approach. PMID- 17415549 TI - The role of physical activity in the management of impaired glucose tolerance: a systematic review. AB - Although physical activity is widely reported to reduce the risk of type 2 diabetes in individuals with prediabetes, few studies have examined this issue independently of other lifestyle modifications. The aim of this review is to conduct a systematic review of controlled trials to determine the independent effect of exercise on glucose levels and risk of type 2 diabetes in people with prediabetes (IGT and/or IFG). A detailed search of MEDLINE (1966-2006) and EMBASE (1980-2006) found 279 potentially relevant studies, eight of which met the inclusion criteria for this review. All eight studies were controlled trials in individuals with impaired glucose tolerance. Seven studies used a multi-component lifestyle intervention that included exercise, diet and weight loss goals and one used a structured exercise training intervention. Four studies used the incidence of diabetes over the course of the study as an outcome variable and four relied on 2-h plasma glucose as an outcome measure. In the four studies that measured the incidence of diabetes as an outcome, the risk of diabetes was reduced by approximately 50% (range 42-63%); as these studies reported only small changes in physical activity levels, the reduced risk of diabetes is likely to be attributable to factors other than physical activity. In the remaining four studies, only one reported significant improvements in 2-h plasma glucose even though all but one reported small to moderate increases in maximal oxygen uptake. These results indicate that the contribution of physical activity independent of dietary or weight loss changes to the prevention of type 2 diabetes in people with prediabetes is equivocal. PMID- 17415550 TI - Genetic association of single nucleotide polymorphisms in endonuclease G-like 1 gene with type 2 diabetes in a Japanese population. AB - AIMS/HYPOTHESIS: In order to identify type 2 diabetes disease susceptibility gene(s) in a Japanese population, we applied a region-wide case-control association test to the 20.4 Mb region between D3S1293 and D3S2319 on chromosome 3p24.3-22.1, supported by linkage to type 2 diabetes and its related traits in Japanese and multiple populations. MATERIALS AND METHODS: We performed a two stage association test using 1,762 Japanese persons with 485 gene-centric, evenly spaced, common single nucleotide polymorphism (SNP) markers with minor allele frequency >0.1. For mouse studies, total RNA was extracted from various organs of BKS.Cg-+Lepr(db)/+Lepr(db) and control mice, and from MIN6, NIH3T3 and C2C12 cell lines. RESULTS: We detected a landmark SNP375 (A/G) (rs2051211, p = 0.000046, odds ratio = 1.33, 95% CI 1.16-1.53) in intron 5 of the endonuclease G-like 1 (ENDOGL1) gene. Systematic dense SNPs approach identified a susceptibility linkage disequilibrium (LD) block of 116.5 kb by |D'|, an LD units map and a critical region of 2.1 kb by r (2) in ENDOGL1. A haplotype-based association test showed that an at-risk haplotype is associated with disease status (p = 0.00001). The expression of ENDOGL1 was rather ubiquitous with relatively abundant expression in the brain and also in a pancreatic islet beta cell line. Mouse Endogl1 expression increased in pancreatic islets of hyperglycaemic BKS.Cg +Lepr(db)/+Lepr(db) mice compared with that in control mice. CONCLUSIONS/INTERPRETATION: Based on the population genetics, fine mapping of LD block and haplotype analysis, we conclude that ENDOGL1 is a candidate disease susceptibility gene for type 2 diabetes in a Japanese population. Further analysis in a larger sample size is required to substantiate this conclusion. PMID- 17415551 TI - Risk of hypoglycaemia in types 1 and 2 diabetes: effects of treatment modalities and their duration. AB - AIMS/HYPOTHESIS: We explored the epidemiology of hypoglycaemia in individuals with insulin-treated diabetes by testing the hypothesis that diabetes type and duration of insulin treatment influence the risk of hypoglycaemia. MATERIALS AND METHODS: This was an observational study over 9-12 months in six UK secondary care diabetes centres. Altogether 383 patients were involved. Patients were divided into the following three treatment groups for type 2 diabetes: (1) sulfonylureas, (2) insulin for <2 years and (3) insulin for >5 years, and into two treatment groups for type 1 diabetes, namely <5 years disease duration and >15 years disease duration. Self-reported (mild and severe) and biochemical episodes (interstitial glucose <2.2 mmol/l using continuous glucose monitoring) were recorded. RESULTS: Mild hypoglycaemia in type 2 diabetic patients on insulin for <2 years was less frequent than in type 1 patients with <5 years disease duration (mean rate: 4 vs 36 episodes per subject-year, p < 0.001). In type 2 diabetic patients treated with sulfonylureas or insulin for <2 years, no differences were observed in the proportion experiencing severe hypoglycaemia (7 vs 7%, difference 0 [95% CI: -7 to 9%]), mild symptomatic (39 vs 51%, difference 12 [-3 to 25%]) or interstitial glucose <2.2 mol/l (22 vs 20%, difference 2 [-13 to 10%]). Severe hypoglycaemia rates were comparable in patients with type 2 diabetes on sulfonylureas or insulin < 2 years (0.1 and 0.2 episodes per subject year) and far less frequent than in type 1 diabetes (<5 years group, 1.1; >15 years group, 3.2.episodes per subject-year). CONCLUSIONS/INTERPRETATION: During early insulin use in type 2 diabetes, the frequency of hypoglycaemia is generally equivalent to that observed in patients treated with sulfonylureas and considerably lower than during the first 5 years of treatment in type 1 diabetes. PMID- 17415552 TI - Do alcohol cues facilitate or impair cognitive processing in recently detoxified alcoholics? Commentary on Noel et al. (2007). PMID- 17415554 TI - Pharmacogenetics of SLCO1B1: haplotypes, htSNPs and hepatic expression in three distinct Asian populations. AB - OBJECTIVE: The aim of this study was to characterize the population frequency of SLCO1B1 polymorphic variants in three distinct healthy Asian populations, namely Chinese (n = 100), Malay (n = 100) and Indian (n = 100), and to explore the association between haplotype-tagged single nucleotide polymorphisms (htSNPs) on hepatic SLCO1B1 mRNA expression. METHODS: The distribution of polymorphic variants in the SLCO1B1 gene at eight loci that spanned approximately 48 kb was investigated in the three different Asian ethnic groups and in 32 non-cancerous liver tissues from Chinese patients. RESULTS: Of the 26 polymorphisms screened, we found eight polymorphic variants that differed in genotypic and allelic frequencies between the Chinese, Malay and Indian populations. Significant interethnic differences were observed in the genotype frequency distributions across the promoter SNP [g.-11187G>A (P = 0.030)] as well as three coding region SNPs [c.388G>A (P < 0.001); c.571T>C (P < 0.001); c.597C>T (P < 0.001)] in the healthy subjects. Haplotype analysis revealed 12 different haplotypes in both the Chinese and Malay populations and 18 haplotypes in the Indian population. In both the Malay and Indian populations, the htSNPs were c.388A>G, c.571T>C and c.597C>T, whereas in the Chinese population they were g.-11187G>A, c.388A>G and c.597C>T. The c.388A>G and c.597C>T htSNPs accounted for more than 70% of the variations between the three major haplotypes in each Asian ethnic group. In terms of the c.388A>G htSNPs, genotypic-phenotypic association analyses revealed that there was no effect on SLCO1B1 expression in hepatic tissues; in addition, no genotypic-phenotypic associations were evident with regards to the c.597C>T htSNP. CONCLUSION: Future studies should investigate the phenotypic effects of the c.388A>G htSNP on the disposition of OATP1B1 substrates in Asian populations. PMID- 17415555 TI - Midazolam anesthesia during rigid and flexible cystoscopy. AB - The objective of this study was to investigate the usefulness and safety of midazolam-induced anesthesia for cystoscopy. From September 2005 to March 2006, 80 patients scheduled for regular outpatient follow-up cystoscopy participated in this study. The patients were randomized and classified according to the cystoscope type and midazolam use as follows: group 1 (10 men and 10 women), flexible cystoscopy + midazolam; group 2 (10 men and 10 women), flexible cystoscopy + no midazolam; group 3 (10 men and 10 women), rigid cystoscopy + midazolam; and group 4 (10 men and 10 women), rigid cystoscopy + no midazolam. Immediately after the procedure, the patients were asked to rate their comfort level using a ten-point visual linear analog self-assessment pain scale. The patients assigned in the midazolam group experienced no side-effects from the midazolam. Blood pressure and pulse rate did not change significantly during the procedure. The degree of pain experienced by group 1 was lower than other groups (P < 0.05) and group 4 had a significantly greater pain score than other groups (P < 0.05). No difference was evident between group 2 and 3 regarding the pain score. Midazolam anesthesia may relieve pain during rigid and flexible cystoscopy. Our findings suggest that midazolam anesthesia during cystoscopy is useful and safe. PMID- 17415556 TI - Modulation of the reaction cycle of the Na+:Ca2+, K+ exchanger. AB - Ca(2+) concentration in retinal photoreceptor rod outer segment (OS) strongly affects the generator potential kinetics and the receptor light adaptation. The response to intense light stimuli delivered in the dark produce potential changes exceeding 40 mV: since the Ca(2+) extrusion in the OS is entirely controlled by the Na(+):Ca(2+), K(+) exchanger, it is important to assess how the exchanger ion transport rate is affected by the voltage and, in general, by intracellular factors. It is indeed known that the cardiac Na(+):Ca(2+) exchanger is regulated by Mg-ATP via a still unknown metabolic pathway. In the present work, the Na(+):Ca(2+), K(+) exchanger regulation was investigated in isolated OS, recorded in whole-cell configuration, using ionic conditions that activated maximally the exchanger in both forward and reverse mode. In all species examined (amphibia: Rana esculenta and Ambystoma mexicanum; reptilia: Gecko gecko), the forward (reverse) exchange current increased about linearly for negative (positive) voltages and exhibited outward (inward) rectification for positive (negative) voltages. Since hyperpolarisation increases Ca(2+) extrusion rate, the recovery of the dark level of Ca(2+) (and, in turn, of the generator potential) after intense light stimuli results accelerated. Mg-ATP increased the size of forward and reverse exchange current by a factor of approximately 2.3 and approximately 2.6, respectively, without modifying their voltage dependence. This indicates that Mg-ATP regulates the number of active exchanger sites and/or the exchanger turnover number, although via an unknown mechanism. PMID- 17415557 TI - Spectral analysis of Delayed Luminescence from human skin as a possible non invasive diagnostic tool. AB - In vivo measurements of Delayed Luminescence (DL), the low-level photo-induced emission which lasts for a longer time after switching off the excitation light, have been performed on human skin, with the aim to develop a technique for optical biopsy. Preliminary tests have been performed on healthy volunteers, measuring the time decays of the spectral components (lambda(emiss) = 400-800 nm) starting 10 mus after switching off the excitation (lambda(exc) = 337 nm). Significant differences in the decay trends of DL from different subjects were revealed and quite a good reproducibility for the same subject was observed. The modeling of experimental data has been examined in detail in order to get parameters, characterizing the theoretical fit, whose changes may be correlated with age differences and seasonal variations. PMID- 17415558 TI - Characterization of two beta-carotene ketolases, CrtO and CrtW, by complementation analysis in Escherichia coli. AB - The pathways from beta-carotene to astaxanthin are crucial key steps for producing astaxanthin, one of industrially useful carotenoids, in heterologous hosts. Two beta-carotene ketolases (beta-carotene 4,4'-oxygenase), CrtO and CrtW, with different structure are known up to the present. In this paper, we compared the catalytic functions of a CrtO ketolase that was obtained from a marine bacterium Rhodococcus erythropolis strain PR4, CrtO derived from cyanobacterium Synechosistis sp. PCC6803, and CrtW derived from a marine bacterium Brevundimonas sp. SD212, by complementation analysis in Escherichia coli expressing the known crt genes. Results strongly suggested that a CrtO-type ketolase was unable to synthesize astaxanthin from zeaxanthin, i.e., only a CrtW-type ketolase could accept 3-hydroxy-beta-ionone ring as the substrate. Their catalytic efficiency for synthesizing canthaxanthin from beta-carotene was also examined. The results obtained up to the present clearly suggest that the bacterial crtW and crtZ genes are a combination of the most promising gene candidates for developing recombinant hosts that produce astaxanthin as the predominant carotenoid. PMID- 17415559 TI - Preparing a highly specific inert immunomolecular-magnetic beads for rapid detection and separation of S. aureus and group G Streptococcus. AB - The rapid detection and separation of Staphylococcus aureus and group G Streptococcus was based on the affinity chromatography interactions between Fc fragment of human IgG and protein A/G (located on the cell wall of S. aureus and group G Streptococcus). In this case, immobilization of antibodies had to take place in a different and complementary way than in the case of conventional immunosensors. In this study, three different kinds of immunomolecular-magnetic beads (IMB) were prepared for rapid detection and separation of S. aureus and group G Streptococcus (GGS). The Fc regions of the immobilized antibodies were fully accessible to adsorb protein A or protein G. On the contrary, conventional immunosensors had to have fully accessible Fab regions to facilitate the antigen antibody recognition. It was suggested that the worse method of immobilization of the antibodies for conventional use would yield the better results for this specific use. In this study, we also perfectly solved the nonspecific adsorptions and interaction problems, which were the most serious critical problems for all kinds of sensors. It was achieved by blocking the excess surface groups of aldehyde IMB and the Fab region of the immobilized antibodies with aldehyde dextran. PMID- 17415560 TI - Re-examining the association of os acromiale with supraspinatus and infraspinatus tears. AB - PURPOSE: To re-evaluate the relationship between os acromiale and rotator cuff tears. METHODS: We retrospectively analyzed 84 magnetic resonance imaging studies of the shoulder. Forty-two subjects with os acromiale (n = 42; 32 men and ten women, age 25-81 years, mean 47.6 years) were compared with age- and gender matched subjects with no evidence of os acromiale (controls). Arthroscopy data were available in 19 os acromiale and 12 control subjects. Statistical analyses were performed to determine differences between groups regarding rotator cuff tears affecting the supraspinatus and infraspinatus tendons detected by magnetic resonance imaging and arthroscopy. Analysis of os acromiale type, ossicle synchondrosis edema, acromioclavicular joint degenerative changes and step-off deformity at the synchondrosis were tabulated. RESULTS: No statistically significant difference between the os acromiale and control groups was noted, either on magnetic resonance imaging or arthroscopy, with regard to tears of the supraspinatus (P = 1.000 and 0.981, respectively) and infraspinatus (P = 1.000 and 0.667, respectively) tendons. There was a statistically significant increased number of supraspinatus (P = 0.007) and infraspinatus (P = 0.03) tears in a comparison of subjects with os acromiale and step-off deformity (10/42) vs os acromiale without step-off deformity (32/42). CONCLUSION: The presence of os acromiale may not significantly predispose to supraspinatus and infraspinatus tendon tears. However, subjects with step-off deformity of an os acromiale are at greater risk of rotator cuff tears than are similar subjects without such deformity. PMID- 17415561 TI - Extracorporeal shock wave treatment for shoulder calcific tendonitis: a systematic review. AB - The treatment of patients with calcific tendonitis is typically conservative, including physical therapy, iontophoresis, deep friction, local or systemic application of noninflammatory drugs, needle irrigation-aspiration of calcium deposit, and subacromial bursal steroid injection. If the pain becomes chronic or intermittent after several months of conservative treatment, arthroscopic and open procedures are available to curette the calcium deposit, and additional subacromial decompression can be performed if necessary. As an alternative, minimally invasive extracorporeal shock wave therapy (ESWT) has been postulated to be an effective treatment option for treating calcific tendinitis of the shoulder, before surgery. Herein we discuss the indications, mechanism of therapeutic effect, efficacy of treatment, and complications after ESWT application. PMID- 17415562 TI - Imaging of pseudoneoplastic masses associated with allografts. AB - OBJECTIVE: The objective of this study is to describe the imaging features of non neoplastic masses suspected of being tumor recurrences adjacent to allografts. The allografts were utilized for the treatment of various musculoskeletal tumors. MATERIALS AND METHODS: We reviewed the medical records and imaging studies of 56 patients who were suspected of having recurrent tumors following surgical resection and allograft replacement treatment for a variety of musculoskeletal neoplasms. The imaging modalities included radiographs, CT, and MRI. RESULTS: There were 47 cases of recurrent tumors. All tumor recurrences were in the soft tissues of the surgical bed (41 patients), or in the native bone adjacent to the host/allograft junction (6 patients). None of the recurrences originated in the allografts. Nine patients suspected of having recurrences were discovered to have non-neoplastic masses. Five of these were very closely related to the allograft, wrapping around parts of the allograft, and the other 4 were in the surgical bed, 2 of which were abscesses and two were seromas. CONCLUSION: Most masses arising in the vicinity of allografts implanted following resection of musculoskeletal tumors represent recurrent neoplasms. A minority are reactive processes or abscesses or fluid collections. These "pseudoneoplasms," specifically those closely related to the allografts, have specific imaging characteristics that help distinguish them from recurrent tumors. PMID- 17415563 TI - Early detection and quantification of murine melanoma bone metastases with magnetic resonance imaging. AB - OBJECTIVE: Bone metastases occur in approximately 80% of patients with advanced cancer and cause significant morbidity. There are currently no established means by which to identify the early growth of micro-metastatic cells or their effects on bone at a time when curative therapy might be initiated. We postulated that high-resolution magnetic resonance imaging (MRI) could detect and quantify the growth and destructive effects of bone micrometastases. DESIGN: Using a mouse model for metastasis of malignant melanoma, we have examined the ability of MRI to quantify cortical bone destruction and the percentage of the medullary cavity occupied by tumour, trabecular bone, and marrow. The results from MRI were compared to histomorphometry (the reference standard) and to radiographs. RESULTS: In vivo gradient-echo and spin-echo MRI demonstrated that metastatic melanoma replaced the marrow space but that the cortical bone integrity was preserved (P < or = 0.001). The smallest detectable micrometastasis had an area of 0.323 mm(2). In contrast, we observed no trends after quantifying the radiograph data. CONCLUSION: These approaches delineated the limits of MRI in its ability to quantify tumour burden and the effect on bone in this model. Given the increasing use of MRI as a non-invasive clinical diagnostic method, the present findings may be applicable in detecting bone metastases in the clinical setting at an early and potentially treatable stage. PMID- 17415565 TI - T cells are crucial for the anti-metastatic effect of anti-epidermal growth factor receptor antibodies. AB - Experimental evidences supporting the epidermal growth factor receptor (EGFR) as an important molecule for tumor metastasis had been accumulated. Currently, anti EGFR monoclonal antibodies (mAbs) constitute a promising approach for the treatment of patients with metastatic tumors. However, the mechanisms associated with the potent anti-metastatic effect of these mAbs have not been completely elucidated due to the lack of appropriate syngeneic preclinical models. In this paper, we have investigated the effects of 7A7, an antibody specific to murine EGFR, on the metastatic properties of D122 murine lung carcinoma. 7A7 mAb significantly impaired metastatic spread of D122 cells in C57BL/6 mice by direct anti-proliferative and pro-apoptotic effects on tumor metastasis. 7A7 mAb capacity to inhibit EGFR activation on D122 cells could contribute to its anti metastatic effect. In addition, 7A7 mAb was able to induce in vitro antibody dependent cell-mediated cytotoxicity on D122 cells. Interestingly, 7A7 mAb treatment increased the number of natural killer cells, T lymphocytes and dendritic cells infiltrating the metastatic sites. More strikingly, depletion of CD8(+) and CD4(+) T cells in vivo completely abrogated the 7A7 mAb anti metastatic activity whereas function of natural killer cells was irrelevant. This study supports an in vivo role for T cell response in the mechanism of action of anti-EGFR mAbs, suggesting the induction of an adjuvant effect. PMID- 17415564 TI - Abdominal visceral fat accumulation is associated with the results of (123)I metaiodobenzylguanidine myocardial scintigraphy in type 2 diabetic patients. AB - PURPOSE: We tested the hypothesis that increased abdominal visceral accumulation (VFA) is associated with insulin resistance and cardiovascular autonomic dysfunction in type 2 diabetic patients not receiving insulin treatment. METHODS: The fat distribution was evaluated by measuring the VFA by abdominal computed tomography at the umbilical level. The study group consisted of 24 type 2 diabetic patients with high VFA (> or =100 cm(2), age 60 +/- 8 years, high VFA group). The control group consisted of 19 age-matched type 2 diabetic patients with normal VFA (<100 cm(2), age 60 +/- 7 years, normal VFA group). Cardiovascular autonomic function was assessed by baroreflex sensitivity, heart rate variability, plasma norepinephrine concentrations, and cardiac (123)I metaiodobenzylguanidine (MIBG) scintigraphy. RESULTS: Early and delayed (123)I MIBG myocardial uptake values were lower (p < 0.005 and p < 0.0001, respectively) and the percent washout rate of (123)I-MIBG was higher (p < 0.0005) in the high VFA group than in the normal VFA group. The fasting plasma insulin concentrations (p < 0.005) and the homeostasis model assessment (HOMA) index values (p < 0.0005) were higher in the high VFA group than in normal VFA group. Multiple regression analysis revealed that the level of VFA was independently predicted by the HOMA index values and the myocardial uptake of (123)I-MIBG during the delayed phase. CONCLUSION: Our results demonstrate that the level of VFA is associated with depressed cardiovascular autonomic function and insulin resistance in patients with type 2 diabetes mellitus. PMID- 17415566 TI - Hyponatremia after administration of conditioning regimen in myelodysplastic syndrome with empty sella after glandula pituitaria surgery. PMID- 17415567 TI - Characterization of genes encoding novel peptidases in the biocontrol fungus Trichoderma harzianum CECT 2413 using the TrichoEST functional genomics approach. AB - Proteolytic enzymes (EC 3.4) secreted by Trichoderma strains are receiving increasing attention because of their potential implication in the Trichoderma biocontrol abilities. We have used an expressed sequence tag (EST) approach to identify genes encoding extracellular peptidases in T. harzianum CECT 2413 grown under several biocontrol-related conditions. Based on BlastX results and Gene Ontology annotation, a total of 61 (among 3478) unique sequences (unisequences) were predicted to encode enzymes with peptidase activity, three corresponding to secreted peptidases already known from this Trichoderma strain (PAPA, PRA1 and P6281). Further manual screening based on the functional identity and cellular location of the best matches revealed ten unisequences encoding novel extracellular peptidases. We report the characterization of the corresponding genes as well as a potential orthologous gene of the intracellular peptidase PAPB from T. asperellum. In each case, full-length coding sequences were obtained, and deduced proteins were compared at phylogenetic level with peptidases from other organisms. T. harzianum CECT 2413 novel peptidases included six serine endopeptidases (EC 3.4.21) belonging to the families S1, S8 and S53, three aspartic endopeptidases (EC 3.4.23) of the family A1, one metallo-endopeptidase (EC 3.4.24) of the family M35, and one aminopeptidase (EC 3.4.11) of the family M28. Results obtained by Northern blot analyses demonstrated that the genes within a family are differentially regulated in response to different culture conditions, suggesting that they have diverse functional roles. PMID- 17415568 TI - Propagation of the [PIN+] prion by fragments of Rnq1 fused to GFP. AB - Prions are viewed as enigmatic infectious entities whose genetic properties are enciphered solely in an array of self-propagating protein aggregate conformations. Rnq1, a yeast protein with yet unknown function, forms a prion named [PIN+] for its ability to facilitate the de novo induction of another prion, [PSI+]. Here we investigate a set of RNQ1 truncations that were designed to cover major Rnq1 sequence elements similar to those important for the propagation of other yeast prions: a region rich in asparagines and glutamines and several types of oligopeptide repeats. Proteins encoded by these RNQ1 truncations were tested for their ability to (a) join (decorate) pre-existing [PIN+] aggregates made of wild-type Rnq1 and (b) maintain the heritable aggregated state in the absence of wild-type RNQ1. While the possible involvement of particular sequence elements in the propagation of [PIN+] is discussed, the major result is that the efficiency of transmission of [PIN+] from wild-type Rnq1 to a fragment decreased with the fragment's length. PMID- 17415569 TI - The symbiosis phenotype and expression patterns of five nodule-specific genes of Astragalus sinicus under ammonium and salt stress conditions. AB - In previous works, we isolated 14 nodule-specific or nodule-enhanced genes from Astragalus sinicus by suppressive subtractive hybridization. In this study, we have further identified the expression patterns of five nodule-specific genes of A. sinicus under salt and ammonium stress. Transcription levels of genes tested were quantified by quantitative fluorescence real-time RT-PCR. Results showed that: (1) About 80 mM NaCl and all stress treatments containing (NH(4))(2)SO(4) significantly inhibited nitrogen-fixing capacity of inoculated plants. About 40 mM NaCl showed relative lighter inhibition. (2) Compare with positive control at normal conditions, the expressions of all genes were significantly reduced by all ammonium stress. (3) Under salt stress without exogenous nitrogen, transcription levels of AsIIA255 and AsE246 were significantly increased after treatment for 3 days. But expressions of AsG2411, AsIIC2512, and AsB2510 were suppressed by 80 mM NaCl and not significantly affected by 40 mM NaCl. (4) Under salt stress with exogenous nitrogen, expressions of AsG2411, AsIIC2512, AsB2510, and AsIIA255 were significantly suppressed. While, the transcription level of AsE246 under 80 mM NaCl containing 1 mM (NH(4))(2)SO(4) was still higher than that of positive control. The correlation of the expression profiles of three cysteine cluster protein (CCP) genes (AsG2411, AsIIC2512, AsIIA255) and one lipid transfer protein (LTP) gene (AsE246) with the nitrogen-fixing capacities of nodules in each treatments may explain the molecular mechanisms of their supposed functions in symbiosis and nitrogen-fixing process. Our results also implied that AsIIA255 and AsE246 might play a role in the response of A. sinicus to salt stress to facilitate the nitrogen-fixation process. PMID- 17415570 TI - Endoscopic third ventriculostomy in children younger than 2 years of age. AB - INTRODUCTION: Endoscopic third ventriculostomy (ETV) for the treatment of hydrocephalus of different etiologies is still controversial in children younger than 2 years of age. The success rate of ETV in this group of patients is analyzed in this study. MATERIALS AND METHODS: The series consisted of 21 patients treated with ETV. The mean age of the patients was 6.7 months, ranging from 9 days to 15 months (16 patients were younger than 1 year). The study included hydrocephalus due to idiopathic aqueductal stenosis (eight) and other congenital anomalies (four) as well as posthemorrhagic (three) and tumor-related occlusive hydrocephalus (three). Two patients presented with shunt infection and one with a shunt failure. ETV was considered to be successful when shunting could be avoided. RESULTS: ETV was successful in nine patients, with a mean follow-up period of 26.2 months. The procedure was successful in four patients with idiopathic aqueductal stenosis, in two with other congenital anomalies, in one posthemorrhagic, and in two with a tumor-related hydrocephalus. In 12 patients, the ETV was unsuccessful after a mean follow-up of 3.3 months. These patients required a shunt. Ten of them were less than 1 year old when ETV was performed. In one tumor-related hydrocephalus, a shunt was inserted after a meningitis after tumor removal. CONCLUSIONS: The success of ETV in children younger than 2 years of age suffering from non-communicating hydrocephalus seems to be dependent on both age and etiology. Our results show an overall success rate of 43%. In 37.5% of the children younger than 1 year of age, ETV was successful. ETV in patients with hydrocephalus due to idiopathic aqueductal stenosis seems to be more beneficial than in other causes of hydrocephalus. PMID- 17415571 TI - The extended endoscopic endonasal approach to the clivus and cranio-vertebral junction: anatomical study. AB - OBJECT: Lesions located in the retroclival area and at the level of the cranio vertebral junction are typically approached through a variety of anterior, antero lateral and postero-lateral skull-based approach, either alone or in combination. The aim of this anatomical study was to demonstrate the possibility of an endoscopic endonasal approach to the clivus and cranio-vertebral junction. MATERIALS AND METHODS: Five fresh cadaver heads injected with colored latex were used. A modified endonasal endoscopic approach was made through two nostrils in all cases. Endoscopic dissections were performed using rigid endoscopes, 4 mm in diameter, 18 cm in length, with 0 degrees lenses. RESULTS: Access to the clivus was possible using a lower trajectory when compared to that necessary for the sellar region. The sphenoid sinus is entered, and its inferior wall is completely removed, permitting the union of the sphenoidal and rhinopharyngeal parts of the clivus. The entire clivus can be removed, and the cranio-vertebral junction is opened, removing the anterior arch of atlas and the odontoid process. After the opening of the dura, the anterior surface of the pons and upper spinal cord with corresponding nerves and vasculature are visible. CONCLUSION: The endoscopic endonasal approach to the clivus and cranio-vertebral junction could be a valid alternative for surgical treatment of anterior lesions of these regions. Adequate endoscopic skill, lab training on cadavers and dedicated tools are required for clinical applications of the approach. PMID- 17415572 TI - Endoscopic management of intra and paraventricular CSF cysts. AB - BACKGROUND: The management of intra- and paraventricular lesions is one of the fields in which modern neuroendoscopic techniques have given the most significant contribution in terms of reduction in operative invasiveness and postoperative complications. In this context, fluid cysts represent an ideal ground on which results obtained with traditional surgical procedures (open surgery, shunting) have more and more to compare with the increasing number of encouraging results obtained by neuroendoscopy. OBJECTIVE: The objective of this study was to retrospectively evaluate the results of the endoscopic treatment of intra- and paraventricular cysts performed at our institution and how they compare with what has been reported in the literature, concerning the results of open surgery, shunting, and endoscopic management. MATERIALS AND METHODS: All the patients with intra- and paraventricular cerebrospinal fluid (CSF) cysts endoscopically managed at our institution between March 2000 and October 2006 were included. Based on cyst location documented by magnetic resonance imaging (MRI), the cohort of patients examined was subdivided in five groups: A, paraventricular cysts; B, choroids plexuses cysts; C, secondary intraventricular cysts in previously hydrocephalus shunted patients; D, quadrigeminal cistern arachnoid cysts; and E, suprasellar arachnoid cysts. The neuroendoscopic procedures were performed with a 30 degrees rigid endoscope (Storz Decq, 3.8 mm). The surgical plan and best trajectory were selected on preoperative MR imaging. Postoperatively, all patients underwent CT scans in the first 48 h after surgery and MR control 3 months after surgery. RESULTS: There were 26 patients (18 M/8F). The mean age at diagnosis was 8.95 years. Five of 26 cases were adult patients. Four patients had paraventricular cysts; 2 patients had a choroids plexuses cyst; 6 patients a secondary intraventricular cyst in previously shunted hydrocephalus; 11 patients had a quadrigeminal cistern arachnoid cyst and 3 patients had a suprasellar arachnoid cyst. Fifteen patients had an associated hydrocephalus at the time of the surgical treatment (one patient in group A, six patients in group C, six in group D, and two in group E). Twenty-three out of 26 patients underwent endoscopic management of their cystic lesion as primary procedure; in the remaining three cases, it represented a secondary procedure after open cyst marsupialization or/and the implant of a cystoperitoneal shunt. Endoscopic management consisted in a cystoventriculostomy (CV) in 19 patients. Third ventriculostomy (ETV) was associated to CV in seven cases, and it was performed at the same time of the CV procedure in all these cases. There was no mortality, neither operative morbidity. At a mean follow-up of 2.15 years (0.1-4 years), a complete resolution of preoperative clinical symptoms and signs was recorded in 80.9% of symptomatic patients. Control MRI showed a reduced cyst size in 25/26 patients and a stable cyst size in the last case, with signs of CSF flow between the cyst and the cerebral ventricles in all cases. One out of 15 patients with associated hydrocephalus had a persistent ventriculomegaly without signs of increased intracranial pressure. CONCLUSIONS: Our results confirm that endoscopic management of intra- and paraventricular cysts is a valid alternative to open surgery as well as to shunting procedures. Control of clinical symptoms and signs was obtained in around 80% of our patients, while radiological evidence of cyst size reduction occurred in more than 95% of them. These rates are comparable with results of open surgery and shunting. The main advantage of neuroendoscopy is the low incidence of complications, a result that is confirmed by the present series. PMID- 17415573 TI - Telovelar approach: technical issues for large fourth ventricle tumors. AB - OBJECTS: Fourth ventricle is conventionally accessed via resection of the part of the vermis for total excision of the tumors at the expense of significant morbidity. Numerous avenues have been identified to minimize the morbidity; some of which include transforaminal, subtonsillar, telovelar approaches, etc. These approaches are devised on the basis that accurate dissection along the natural avascular planes will avoid injury to the important structures in this area minimizing morbidity. We attempt to emphasize the technique of telovelar approach and the problems encountered while employing this technique for excision of large fourth ventricle tumors. MATERIALS AND METHODS: Fifteen patients with fourth ventricle tumors were operated during January to September 2005 at Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala, India. Fourteen of these cases were medulloblastomas, and one was ependymoma. All the patients were operated in prone oblique position via telovelar approach. CONCLUSION: Clear understanding of the normal anatomy will help in meticulous dissection and will result in reduced morbidity. Significant incidence of postoperative ataxia and mutism is seen with this approach in large tumors, and this can be avoided by staged dissection of the uvulotonsillar cleft. PMID- 17415574 TI - Absence of heterogeneous nuclear ribonucleoproteins and survival motor neuron protein in TDP-43 positive inclusions in frontotemporal lobar degeneration. AB - TDP-43 was recently identified as the major disease protein in neuronal inclusions in frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). TDP-43 becomes redistributed from the nucleus to the cytoplasm, ubiquitinated, hyperphosphorylated and cleaved to generate C-terminal fragments, thereby linking mismetabolism of TDP-43 to the pathogenesis of FTLD-U. The function of TDP-43 is unclear, however it has been shown that TDP-43 might act as transcription repressor and activator of exon skipping through interaction with proteins of the heterogeneous nuclear ribonucleoprotein (hnRNP) family as well as a scaffold for nuclear bodies through interactions with survival motor neuron protein. To investigate whether these binding partners might be associated with TDP-43 pathology, we studied the expression and localization of proteins of the hnRNP family (hnRNP A1, A2/B1, C1/C2) and SMN protein in affected brain regions in patients with sporadic and familial FTLD-U and normal control brains by immunohistochemistry and biochemical analysis. In contrast to TDP-43, no changes in subcellular distribution, no labeling of pathologic inclusions and no biochemical alterations were detectable for the tested hnRNPs and SMN in FTLD-U brains compared to controls. These results argue against a role of these binding partners in the pathogenesis of FTLD-U and emphasize the specificity of TDP-43 as marker for FTLD-U pathology. PMID- 17415575 TI - Netherton syndrome: mutation analysis of two Taiwanese families. AB - Netherton syndrome (NS) is a severe autosomal recessive skin disorder characterized by congenital ichthyosiform erythroderma, hair shaft abnormalities, and atopic diathesis. Recently, pathogenic mutations were identified in serine protease inhibitor Kazal-type 5 (SPINK5), the gene that encodes lympho-epithelial Kazal-type related inhibitor (LEKTI), a type of serine protease inhibitor involved in the regulation of skin barrier formation and immunity. In the present report, we describe the mutation analysis of two Taiwanese patients with NS. Patient 1 has heterozygous mutations; the maternal allele has novel T808I (C-T transition in codon 808) and the paternal allele has recurrent R790X (C-T transition in codon 790). Patient 2 is homozygous for a novel polymorphism R267Q (G-A transition in codon 267). The change was not detected in the patient's father. Haplotype analysis revealed that the patient was homozygous for the 5 single nucleotide polymorphisms in the genomic sequence of SPINK5 as well as the flanking (GT)(17) and D5S413, in addition to the discrepancy of R267Q. Nevertheless real-time quantitative PCR analysis revealed no microdeletion in the genomic sequence of SPINK5. Thus uniparental disomy of maternal SPINK5 allele was indicated. PMID- 17415576 TI - Expression of human beta-defensins in patients with mycosis fungoides. AB - The human beta-defensins (hBDs) are peptides with a strong antimicrobial activity. Patients with atopic dermatitis (AD) and mycosis fungoides (MF) are prone to skin infections. We aimed to investigate the mRNA expression of hBDs in lesional and non-lesional skin of MF patients, and to compare the data with hBD levels found in AD patients and healthy controls. In this prospective pilot study, 13 patients with MF were recruited. Punch biopsies were harvested from the centre of the tumour (lesional) as well as a healthy skin site (non-lesional controls). In addition to the specimens of MF patients, skin samples (healthy controls) were obtained from healthy subjects (n = 15) and patients with acute AD (n = 14). In order to detect mRNA of hBDs, we performed quantitative real-time reverse transcriptase polymerase chain reaction. As compared to healthy controls, skin of patients with MF (non-lesional and lesional) and AD patients showed significantly lower hBD-1 mRNA expression and significantly higher hBD-2 and hBD 3 mRNA expression. HBD-1 mRNA levels of lesional skin were significantly lower than those of non-lesional skin. By contrast, significantly increased hBD-2 and hBD-3 mRNA expression was found in lesional skin of MF patients when compared to non-lesional skin. HBD mRNA expression in lesional skin of MF patients did not significantly differ from hBD expression that was observed in AD lesions. We observed an identical pattern of hBD expression in MD and AD suggesting a common regulatory mechanism that might mainly be driven by T helper 2 lymphocytes. PMID- 17415577 TI - UV-light induced radiation recall dermatitis after a chemoradiotherapy organ preservation protocol. AB - This case report documents a UV-light-induced recall phenomenon and reviews the medical literature. In this patient, we observed a severe recall phenomenon precipitated by an extensive sunlight exposure after a chemoradiotherapy organ preservation protocol. Radiation recall phenomenon is a rare but well-described phenomenon, without clear radiation or drug-specific characteristics. In the medical literature, radiation recall following UV-sunlight exposure seems to be an exceptional event. The etiology remains unknown but could involve local hypersensitivity through a non-immune activation of inflammatory pathways. Due to the increasing number of patients with head and neck malignancies managed with induction chemotherapy and/or chemoradiation organ preservation protocols, the otorhinolaryngologist--head and neck surgeons as well as the radiation therapist should become aware and familiar with this phenomenon. PMID- 17415578 TI - Activity of chemotherapy in the palliative treatment of salivary gland tumors: review of the literature. AB - Chemotherapy is sometimes used as palliation for patients with salivary gland tumors not amenable to curative treatment. However, if chemotherapy improves survival is unknown. To identify some prognostic parameters in this disease, we conducted an extensive review of the literature. The prognostic value of the baseline clinical characteristics and the different chemotherapy regimens used was assessed using the Cox proportional hazards regression model on the available individual data. In addition, the effect of the different chemotherapy drugs on median survival time was evaluated using meta-weighted linear regression with dummy covariates referring to each chemotherapy drug. The total number of patients included in these studies that fit our inclusion criteria was 264 patients. By reviewing carefully the papers and by contacting the different authors, we were able to retrieve the individual data of 205 patients. In the multivariate Cox analysis, only the use of platinum-based chemotherapy was identified as an independent predictor of an increased survival (P = 0.01). These results were confirmed in a meta-analysis where median survival was increased by 2.5 (95% IC:0.7-4.4) and 4.9 (95% IC:0.45-9.4) months for patients treated with platinum (P = 0.007) and anthracyclin-based (P = 0.03) chemotherapy, respectively. Although exploratory, our analysis suggests that treatment with a platinum-based chemotherapy regimen may be an independent factor of better survival for patients with incurable salivary gland neoplasms. PMID- 17415579 TI - Applicability of autologous incus: the influence of age and localization of cholesteatoma. AB - There is a correlation between macroscopic and histologic changes of incus in chronic otitis with cholesteatoma. The incidence and types of damage of auditory ossicles in chronic otitis media were extensively studied, but except some studies on reoperations there are no morphometric studies of human auditory ossicles in chronic otitis media. In order to define the rate of ossicular applicability for tympanoplasties in different macroscopic forms of damage, we analysed histologic changes of human incudes in correlation to their macroscopic alterations. Histomorphometric study of 82 intraoperatively removed human incudes, from chronic otitis media with cholesteatoma (attic, sinus, tensa cholesteatoma), was performed. They were divided according to the age of patients and localization of cholesteatoma. Erosions, vitality, vascular areas, osteomyelitic zones, and applicability of ossicles were measured. Progressive increase of erosions, decrease of vitality, dilatation of vascular spaces and osteomyelitic areas, and decrease of rate of potential applicability for tympanoplasties were found. Residual osteomyelitis was present in incudes with advanced external osteitis and ossicular defect. Macroscopic changes of incudes in cholesteatoma and their histologic structure correlate. Localisation of cholesteatoma is important for ossicular damage. Defects of long process with external osteitis of incudes and more intense pathological changes demand their removal and use of other reconstructive materials for tympanoplasties in order to avoid tympanoplasty failure. PMID- 17415580 TI - Bone anchored hearing aids: a second fixture reduces auditory deprivation in Cambridge. AB - Bone anchored hearing aids are well established for canal atresia, otosclerosis and chronic suppurative otitis media. Refinements in technique to maximise gain while keeping the complications to a minimum are desirable. This study was taken up in order to explore the potential advantage of a second or spare fixture placed at the time of primary surgery. A group of patients who underwent BAHA insertion at The Emmeline Centre for Cochlear Implants and Bone Anchored Hearing Aids, Addenbrookes hospital, Cambridge, UK with the placement of a spare fixture between 1999 and 2002 were compared to those patients with one fixture BAHA undertaken from 1991. Main outcome measures were complications encountered and duration of disability, (i.e. loss of hearing while waiting for new fixture placement). Both groups had similar incidence of complications, but the group with two fixtures suffered a shorter period of disability when a fixture failed. In our experience the use of second or spare fixture reduces the duration of disability. It engenders no additional complications with minimal extra cost. PMID- 17415581 TI - Long-term results in patients after combined laser total arytenoidectomy with posterior cordectomy for bilateral vocal cord paralysis. AB - The problem of ventilation efficiency after total laser arytenoidectomy with posterior cordectomy due to bilateral vocal cord paralysis is discussed. There are a number of views on the usefulness and efficacy of different surgical procedures aimed at widening the glottis, but the studies concerning the long term functional results are still lacking. The objective of the study is to evaluate the durability of ventilation results in patients after laser arytenoidectomy with posterior cordectomy based on the comparison between the early postoperative results and those assessed after a period of 5 years. Thirty patients (24 females, 6 males) aged between 30 and 80 (mean 58.5) with bilateral vocal cord paralysis after thyroid surgery, who underwent laser arytenoidectomy with posterior cordectomy, were analyzed. Ventilation tests were performed immediately after the operation and 5 years later with Body-Master Laab (Jaeger). The actual and predicted values of FVC, FEF(25), FEF(50), FEF(75), PEF, MMEF(75/25), AREA(Ex), sR(T0T), R(T0T) were compared and analyzed. Also, the values of the inspiratory parameters FIV(1), FIF(50), PIF and the coefficients FEF(50)/FIF(50) and FIV(1)/FEV(1) were assessed. Moreover, subjective evaluation was performed based on a questionnaire. The statistical analysis reveals a significant decrease in FIV(1), FIF(50), PIF and FIV(1)/FEV(1). FEF(50)/FIF(50) and sR(T0T) (actual and predicted values) increased significantly after 5 years from the operation. However, the patients did not complain nor was this tendency reflected in their answers to the questionnaire. The probable reasons for such ventilation results are discussed. The role of phoniatric rehabilitation and the time elapsed is emphasized. In general, it can be concluded that laser arytenoidectomy with posterior cordectomy is a durable and effective procedure, although the objective assessment does not fully match the patient's subjective impression. PMID- 17415582 TI - Peculiarities of the effect of low-dose-rate radiation simulating high-altitude flight conditions on mice in vivo. AB - In the present work, the effect of a low-dose rate of high-LET radiation in polychromatic erythrocytes of mice bone marrow was investigated in vivo. The spectral and component composition of the radiation field used was similar to that present in the atmosphere at an altitude of about 10 km. The dose dependence, adaptive response, and genetic instability in the F1 generation born from males irradiated under these conditions were examined using the micronucleus test. Irradiation of the mice was performed for 24 h per day in the radiation field behind the concrete shield of the Serpukhov accelerator. Protons of 70 GeV were used over a period of 15-31 days, to accumulate doses of 11.5-31.5 cGy. The experiment demonstrated that irradiation of mice in vivo in this dose range leads to an increase in cytogenetic damage to bone marrow cells, but does not induce any adaptive response. In mice pre-irradiated with a dose of 11.5 cGy, an increase in sensitivity was observed after an additional irradiation with a dose of 1.5 Gy. The absence of an adaptive response suggests existence of genetic instability. PMID- 17415583 TI - Effects of ageing on the fine distribution of the circadian CLOCK protein in reticular formation neurons. AB - Many biochemical, physiological and behavioural processes, from bacteria to human, exhibit roughly 24 h cyclic oscillations defined as circadian rhythms. However, during ageing, numerous aspects of the circadian biology undergo alterations; in particular, the sleep pattern changes, with more frequent awakenings and shorter sleep time. The basic mechanism of the circadian clock relies on intracellular molecular pathways involving interlocking transcriptional/translational feedback loops, and CLOCK protein, a transcription factor, is essential for normal circadian rhythms. In this study, the fine distribution of CLOCK protein has been analysed, in adult and old rats, at different phases of the daily cycle in the neurons of the medullary reticular formation, involved in the control of the sleep-wake cycle. The results demonstrate quali-quantitative modifications of CLOCK protein in the neurons of old animals, suggesting that such a deregulation of the intracellular clock mechanism may play some role in the degeneration of the sleep-wake circadian cycle. PMID- 17415584 TI - Improved antigen retrieval in freeze-fracture cytochemistry by evaporation of carbon as first replication layer. AB - The recently developed freeze-fracture replica immunolabeling technique uses sodium dodecyl sulfate to clean replicas obtained from chemically unfixed, rapidly frozen cells by evaporation of platinum as first and carbon as second replication layer. The detergent dissolves remains of cellular material with the exception of components which are in direct contact to the replica film. Membrane lipids and membrane protein complexes of the protoplasmic and the exoplasmic membrane halves remain attached to the replica film and are accessible for cytochemical localization. We immunolabeled the membrane proteins caveolin-1 and connexin 43 in mouse cell lines as well as the membrane attached protein tetrachloroethene reductive dehalogenase (PceA) in bacterial cells at freeze fracture replicas generated by different evaporation parameters. The labeling experiments for caveolin-1 and the PceA showed that freeze-fracture replication of cellular membranes accomplished with thin platinum layers as well as replication with carbon as first evaporation layer lead in these cases to an improved antigen retrieval, whereas the labeling efficiency of connexin 43 was not affected by different evaporation conditions. PMID- 17415585 TI - Appraised leadership styles, psychosocial work factors, and musculoskeletal pain among public employees. AB - OBJECTIVES: The main aim of this study was to explore the associations between appraised leadership styles, psychosocial work factors and musculoskeletal pain among subordinates in four different public service sectors from an epidemiological perspective. METHODS: A cross-sectional questionnaire study was conducted; data from 2,403 public sector employees in subordinate positions (86% women) were analysed. The appraised leadership styles were measured through items from a modified version of the CPE questionnaire (C change, P production/structure, E employee/relation). The structure validity of the CPE model was examined by principal component analysis (PCA). Univariate and multivariate analyses of associations between levels of musculoskeletal pain and appraised leadership styles and with psychosocial work factors were conducted. Odds ratios (ORs) with confidence intervals (CIs) of 95% were used as a measure of associations. RESULTS: There were small variations in the appraisals of the immediate manager among the subordinates. However, the associations between musculoskeletal pain and leadership styles varied according to sector. Poor appraisals (low scores) on "change" and "employee relation" dimensions were associated with high levels of musculoskeletal pain in two sectors: home and health care services. In the domestic catering services, poor appraisals of managers in the "production/structure" dimension had the strongest association with high levels of pain. In general, poor appraisals of the "change" dimension was most strongly associated with high levels of musculoskeletal pain. "High work demands" had the strongest association with high levels of pain, particularly among the men. CONCLUSIONS: Poor appraisals of managers and their leadership styles were associated with high levels of musculoskeletal pain among both female and male subordinates in different public service sectors. There is therefore a great need of further studies of the mechanisms behind the relationships between the leadership styles and their impact on health among the genders. PMID- 17415586 TI - Spike-timing-dependent plasticity for neurons with recurrent connections. AB - The dynamics of the learning equation, which describes the evolution of the synaptic weights, is derived in the situation where the network contains recurrent connections. The derivation is carried out for the Poisson neuron model. The spiking-rates of the recurrently connected neurons and their cross correlations are determined self- consistently as a function of the external synaptic inputs. The solution of the learning equation is illustrated by the analysis of the particular case in which there is no external synaptic input. The general learning equation and the fixed-point structure of its solutions is discussed. PMID- 17415587 TI - Identification of novel small RNAs in tomato (Solanum lycopersicum). AB - To date, the majority of plant small RNAs (sRNA) have been identified in rice, poplar and Arabidopsis. To identify novel tomato sRNAs potentially involved in tomato specific processes such as fruit development and/or ripening, we cloned 4,018 sRNAs from tomato fruit tissue at the mature green stage. From this pool of sRNAs, we detected tomato homologues of nine known miRNAs, including miR482; a poplar miRNA not conserved in Arabidopsis or rice. We identified three novel putative miRNAs with flanking sequence that could be folded into a stem-loop precursor structure and which accumulated as 19-24nt RNA. One of these putative miRNAs (Put-miRNA3) exhibited significantly higher expression in fruit compared with leaf tissues, indicating a specific role in fruit development processes. We also identified nine sRNAs that accumulated as 19-24nt RNA species in tomato but genome sequence was not available for these loci. None of the nine sRNAs or three putative miRNAs possessed a homologue in Arabidopsis that had a precursor with a predicted stem-loop structure or that accumulated as a sRNA species, suggesting that the 12 sRNAs we have identified in tomato may have a species specific role in this model fruit species. PMID- 17415588 TI - The P239S palladin variant does not account for a significant fraction of hereditary or early onset pancreas cancer. AB - The P239S palladin variant has recently been suggested to play a role in hereditary pancreatic cancer. We estimated the contribution of the P239S variant, and surrounding sequence, to familial and early-onset pancreatic cancer. The P239S germline variant was identified in one of 84 high-risk cases and one of 555 controls. The case reported an elderly relative with pancreas cancer. We conclude that this variant does not appear to account for a significant fraction of hereditary or early-onset pancreas cancer. PMID- 17415589 TI - Genetic variation in the effect of a facultative symbiont on host-plant use by pea aphids. AB - Ecological specialisation on different host plants occurs frequently among phytophagous insects and is normally assumed to have a genetic basis. However, insects often carry microbial symbionts, which may play a role in the evolution of specialisation. The bacterium Regiella insecticola is a facultative symbiont of pea aphids (Acyrthosiphon pisum) where it is found most frequently in aphid clones feeding on Trifolium giving rise to the hypothesis that it may improve aphid performance on this plant. A study in which R. insecticola was eliminated from a single naturally infected aphid clone supported the hypothesis, but a second involving two aphid clones did not find the same effect. We created a series of new pea aphid-R. insecticola associations by injecting different strains of bacteria into five aphid clones uninfected by symbionts. For all aphid clones, the bacteria decreased the rate at which aphids accepted Vicia faba as a food plant and reduced performance on this plant. Their effect on aphids given Trifolium pratense was more complex: R. insecticola negatively affected acceptance by all aphid clones, had no effect on the performance of four aphid clones, but increased performance of a fifth, thus demonstrating genetic variation in the effect of R. insecticola on pea aphid host use. We discuss how these results may explain the distribution and frequency of this symbiont across different aphid populations. PMID- 17415590 TI - The influence of life form on carbon and nitrogen relationships in tropical rainforest ferns. AB - Tropical ferns are characterized by a high diversity of plant life forms, yet there have been few large-scale studies on the functional ecology of these different forms. We examined epiphytic, hemiepiphytic, and terrestrial ferns, and asked whether there are differences in the mineral nutrition and water relations across different growth forms of a diverse assemblage of species. We measured specific leaf area, leaf nitrogen concentrations, and natural abundance of the stable isotopes delta(15)N and delta(13)C of 48 fern species from 36 genera across a wide range of habitats at La Selva Biological Station in Costa Rica. We found that epiphytes were significantly different in all measured variables from hemiepiphytic and terrestrial species, and that terrestrial and soil-rooted hemiepiphytes were indistinguishable in all variables excluding SLW. A multivariate analysis revealed that aspects of N nutrition were the most reliable at separating epiphytic species from other life forms. Our study demonstrates that the natural abundance of both C and N as well as N relations and leaf morphology are useful when segregating different plant life forms, and that the N cycle of epiphytic and terrestrial habitats function independently from each other. PMID- 17415591 TI - Genetic composition of resident populations influences establishment success of immigrant species. AB - We conducted an outdoor container experiment to test the hypothesis that the genetic composition of resident populations influences the establishment success of immigrant species. We manipulated the genetic compositions (source populations) of populations of the water flea Daphnia magna, a strong competitor in pond and shallow lake zooplankton communities, and monitored the establishment success of immigrant cladoceran species of a regional species pool. We show that establishment success is affected by the source population of the resident D. magna as well as by the presence/absence of macrophytes and the presence/absence of fish in the containers. Our results provide evidence that the genetic composition of resident populations can impact community assembly and metacommunity dynamics, and that community genetics can influence ecosystem functioning. PMID- 17415592 TI - The origin and development of individual size variation in early pelagic stages of fish. AB - Size variation among individuals born at the same time in a common environment (within cohorts) is a common phenomenon in natural populations. Still, the mechanisms behind the development of such variation and its consequences for population processes are far from clear. We experimentally investigated the development of early within-cohort size variation in larval perch (Perca fluviatilis). Specifically we tested the influence of initial variation, resulting from variation in egg strand size, and intraspecific density for the development of size variation. Variation in egg strand size translated into variation in initial larval size and time of hatching, which, in turn, had effects on growth and development. Perch from the smallest egg strands performed on average equally well independent of density, whereas larvae originating from larger egg strands performed less well under high densities. We related this difference in density dependence to size asymmetries in competitive abilities leading to higher growth rates of groups consisting of initially small individuals under high resource limitation. In contrast, within a single group of larvae, smaller individuals grew substantially slower under high densities whereas large individuals performed equally well independent of density. As a result, size variation among individuals within groups (i.e. originating from the same clutch) increased under high densities. This result may be explained by social interactions or differential timing of diet shifts and a depressed resource base for the initially smaller individuals. It is concluded that to fully appreciate the effects of density-dependent processes on individual size variation and size-dependent growth, consumer feedbacks on resources need to be considered. PMID- 17415593 TI - Brown bear body mass and growth in northern and southern Europe. AB - We tested six hypotheses to explain expected geographical differences in body masses of 1,771 brown bears (Ursus arctos) from northern and southern Europe (Sweden and Norway compared with Slovenia and Croatia): Bergmann's rule, the fasting endurance hypothesis, and the dietary meat hypothesis, which predicted larger bears in the north; and hypotheses stressing the role of high primary productivity, high population density, low seasonality, and length of the growing season, which predicted larger bears in the south. Although brown bear populations in North America vary greatly in body mass, we found no significant difference in body mass between the two European populations using a new analytical approach incorporating modeled age-standardized body masses in linear models, when correcting for sex and season. The greater variation in North America may be due primarily to the presence of large bears that feed on salmon (Oncorhynchus spp.), which does not occur in Europe. Asymptotic body masses were 115 +/- 9 (SE) kg in spring and 141 +/- 9 kg in autumn for southern females, 248 +/- 25 and 243 +/- 24 kg for southern males, 96 +/- 2 and 158 +/- 4 kg for northern females, and 201 +/- 4 and 273 +/- 6 kg for northern males, respectively. Northern bears gained more body mass before hibernation and lost more during hibernation than southern bears, probably because hibernation was twice as long in the north. Northern bears gained and southern bears lost mass during the spring, perhaps due to the greater availability and use of protein rich food in spring in the north. As reproductive success in bears is correlated with adult female body mass in interpopulation comparisons, brown bears may have relatively similar reproductive rates throughout Europe, although minimum age at primiparity and litter interval are lower in the south. PMID- 17415594 TI - Resistance to rust fungi in Lolium perenne depends on within-species variation and performance of the host species in grasslands of different plant diversity. AB - The hypothesis that plant species diversity and genetic variation of the host species decrease the severity of plant diseases is supported by studies of agricultural systems, but experimental evidence from more complex systems is scarce. In an experiment with grassland communities of varying species richness (1, 2, 4, 8, 16, and 60 species) and functional group richness (1, 2, 3, and 4 functional groups), we used different cultivars of Lolium perenne (perennial ryegrass) to study effects of biodiversity and cultivar identity on the occurrence and severity of foliar fungal diseases caused by Puccinia coronata (crown rust) and P. graminis (stem rust). Cultivar monocultures of perennial ryegrass revealed strong differences in pathogen susceptibility among these cultivars. Disease intensity caused by both rust fungi decreased significantly with growing species richness of species mixtures. The response to the diversity gradient was related to the decreased density and size of the host individuals with increasing species richness. The occurrence of other grass species known to be possible hosts of the pathogens in the experimental mixtures did not promote disease intensity in L. perenne, indicating that there was a high host specificity of pathogen strains. Differences in pathogen susceptibility among perennial ryegrass cultivars persisted independent of diversity treatment, host density and host individual size, but resulted in a cultivar-specific pattern of changes in pathogen infestation across the species-richness gradient. Our study provided evidence that within-species variation in pathogen susceptibility and competitive interactions of the host species with the environment, as caused by species diversity treatments, are key determinants of the occurrence and severity of fungal diseases. PMID- 17415595 TI - Effectiveness of conservative treatments for the lumbosacral radicular syndrome: a systematic review. AB - Patients with a lumbosacral radicular syndrome are mostly treated conservatively first. The effect of the conservative treatments remains controversial. To assess the effectiveness of conservative treatments of the lumbosacral radicular syndrome (sciatica). Relevant electronic databases and the reference lists of articles up to May 2004 were searched. Randomised clinical trials of all types of conservative treatments for patients with the lumbosacral radicular syndrome selected by two reviewers. Two reviewers independently assessed the methodological quality and the clinical relevance. Because the trials were considered heterogeneous we decided not to perform a meta-analysis but to summarise the results using the rating system of levels of evidence. Thirty trials were included that evaluated injections, traction, physical therapy, bed rest, manipulation, medication, and acupuncture as treatment for the lumbosacral radicular syndrome. Because several trials indicated no evidence of an effect it is not recommended to use corticosteroid injections and traction as treatment option. Whether clinicians should prescribe physical therapy, bed rest, manipulation or medication could not be concluded from this review. At present there is no evidence that one type of treatment is clearly superior to others, including no treatment, for patients with a lumbosacral radicular syndrome. PMID- 17415596 TI - Structural analysis of Cu(II) ligation to the 5'-GMP nucleotide by pulse EPR spectroscopy. AB - Simple copper salts are known to denature poly d(GC). On the other hand, copper complexes of substituted 1,4,7,10,13-pentaazacyclohexadecane-14,16-dione are able to convert the right-handed B form of the same DNA sequence to the corresponding left-handed Z form. A research program was started in order to understand why Cu(II) as an aquated ion melts DNA and induces the conformational change to Z-DNA in the form of an azamacrocyclic complex. In this paper, we present a continuous wave and pulse electron paramagnetic resonance study of the mononucleotide model system Cu(II)-guanosine 5'-monophosphate . Pulse EPR methods like electron nuclear double resonance and hyperfine sublevel correlation spectroscopy provide unique information about the electronic and geometric structure of this model system through an elaborate mapping of the hyperfine and nuclear quadrupole interactions between the unpaired electron of the Cu(II) ion and the magnetic nuclei of the nucleotide ligand. It was found that the Cu(II) ion is directly bound to N7 of guanosine 5'-monophosphate and indirectly bound via a water of hydration to a phosphate group. This set of experiments opens the way to more detailed structural characterization of specifically bound metal ions in a variety of nucleic acids of biological interest, in particular to understand the role of the metal-(poly)nucleotide interaction. PMID- 17415597 TI - Ion conductance vs. pore gating and selectivity in KcsA channel: modeling achievements and perspectives. AB - KcsA potassium channel belongs to a wide family of allosteric proteins that switch between closed and open states conformations in response to a stimulus, and act as a regulator of cation activity in living cells. The gating mechanism and cation selectivity of such channels have been extensively studied in the literature, with a revival emphasis these latter years, due to the publication of the crystallized structure of KcsA. Despite the increasing number of research and review papers on these topics, quantitative interpretation of these processes at the atomic scale is far from achieved. On the basis of available experimental and theoretical data, and by including our recent results, we review the progresses in this field of activity and discuss the weaknesses that should be corrected. In this spirit, we partition the channel into the filter, cavity, extra and intracellular media, in order to analyze separately the specificity of each region. Special emphasis is brought to the study of an open state for the channel and to the different properties generated by the opening. The influence of water as a structural and dynamical component of the channel properties in closed and open states, as well as in the sequential motions of the cations, is analyzed using molecular dynamics simulations and ab initio calculations. The polarization and charge transfer effects on the ions' dynamics and kinetics are discussed in terms of partial charge models. PMID- 17415598 TI - Evaluation of autism traits in Angelman syndrome: a resource to unfold autism genes. AB - Linkage and cytogenetics studies have found the Angelman syndrome (AS) chromosomal region to be of relevance to autism disorder (AD) or autism spectrum disorder (ASD). Autism is considered part of the behavioural phenotype in AS based on formal autism assessments (autism diagnostic interview-revised [ADI-R] and autism diagnostic observation schedule [ADOS]), which have mainly addressed the deleted AS group. We explored 23 AS patients including all genetic subtypes and made a co-morbid diagnosis of AD/ASD in 14/23 (61%), which does not include 4 cases classified within the broader autism spectrum disorder (bASD). Deletions accounted for the main fraction (35%), ubiquitin-protein ligase E3A (UBE3A) mutation represented 13%, imprinting defects and uniparental disomy 9 and 4%, respectively. UBE3A mutations due to lack of the homologous to the E6-associated protein carboxyl terminus domain (n = 3) were associated with the ASD, while more distal mutations (n = 3) seem to escape from a co-morbid diagnosis of autism/autism spectrum. Differences in severity of autistic features were seen across subtypes of AS, with some behavioural features being unique to AS and some representing all forms of developmental disability. Autism signs (poor/lack of eye contact, showing, spontaneous initiation of joint attention, social quality of overtures [ADOS algorithm items for Diagnostic and Statistical Manual of Mental Disorders-IV (DSM-IV)/International Statistical Classification of Diseases and Related Health Problems-10 (ICD-10) autism diagnosis belonging to the reciprocal social interaction domain]) discriminating all the co-morbid AS categories from non-autistic AS belonged to the social interaction domain. Impairments in the communication domain (gestures, pointing, use of another's body, frequency of vocalisation towards others [ADOS algorithm items for DSM IV/ICD-10 autism diagnosis belonging to the communication domain]) justified classification of co-morbid AD/ASD vs the classification of less affected bASD. Evaluation of the behaviour domain suggested that repetitive sensory and motor behaviours correlate with a low developmental profile rather than being specific to autism. PMID- 17415599 TI - Behavioural evidence for separating components within visuo-spatial working memory. AB - Several different sources of evidence support the idea that visuo-spatial working memory can be segregated into separate cognitive subsystems. However, the nature of these systems remains unclear. Recently we reported data from neurological patients suggesting that information about visual appearance is retained in a different subsystem from information about spatial location. In this paper we report latency data from neurologically intact participants showing an experimental double dissociation between memory for appearance and memory for location. This was achieved by use of a selective dual task interference technique. This pattern provides evidence supporting the segregation of visuo spatial memory between two systems, one of which supports memory for stimulus appearance and the other which supports memory for spatial location. PMID- 17415600 TI - Doppler waveforms of the ureteric jet: an overview and implications for the presence of a functional sphincter at the vesicoureteric junction. AB - This paper is a comprehensive review of the Doppler waveform appearance of ureteric jets. Six jet waveform patterns have been identified: monophasic, biphasic, triphasic, polyphasic, square and continuous. Details of the physical properties of jet patterns and their changes under various physiological conditions are illustrated. The immature monophasic ureteric jet pattern is common in infancy and early childhood up to around 4 years of age. This pattern is also noted to have a high incidence in older children with urinary tract infection/vesicoureteric reflux, nocturnal enuresis and in other special physiological conditions such as in children undergoing general anaesthesia, in women during pregnancy, and in patients who have had ureteric transplantation. A hypothesis of dual myogenic and neurogenic components is proposed to explain the mode of action of the vesicoureteric junction (VUJ). The implication of this hypothesis is that it alters the scientific basis of the understanding of the VUJ. Furthermore, the application of colour Doppler US to ureteric jets may provide a non-invasive technique to study the physiology or pathophysiology of the VUJ in humans. This might shed light on new novel approaches to the monitoring and treatment of diseases related to VUJ function. PMID- 17415601 TI - US findings of metacarpophalangeal joints in children with idiopathic juvenile arthritis. AB - BACKGROUND: Juvenile idiopathic arthritis (JIA) is the most common cause of chronic arthritis in children, with frequent involvement of the metacarpophalangeal joints (MCPJ). OBJECTIVE: To compare US findings with those of radiography and clinical examination. MATERIALS AND METHODS: All MCPJs in 20 children with JIA (17 females, median age 9.7 years, range 3.6 to 16.8 years) were evaluated clinically and imaged with gray-scale and color Doppler US, and 90 MCPJs were also imaged radiographically. Each MCPJ was graded on physical examination from 0 (normal) to 4 (severe) by the patient's rheumatologist. RESULTS: US demonstrated abnormalities in 64 of 200 MCPJs (32.0%), including pannus vascularity and/or tenosynovitis in 55 joints (27.5%) (pannus vascularity in 43, tenosynovitis in 40) and bone destruction in 25 joints (12.5%). Overall, US abnormalities and physical examination scores were significantly associated (P < 0.001). However, interobserver agreement between US and clinical evaluation was poor (kappa 0.1) and between US and radiography was only fair (kappa 0.4). CONCLUSION: US of the MCPJ in children with JIA can demonstrate cartilage thinning, bone erosions, and pannus vascularity. Abnormal US findings are significantly correlated with severity of disease as evaluated clinically. PMID- 17415602 TI - Priapism in children: treatment with embolotherapy. AB - BACKGROUND: Priapism is defined as involuntary, prolonged penile erection caused by factors other than sexual arousal, and is classified as either low-flow or high-flow. Embolotherapy is an accepted form of therapy in adults with high-flow priapism. Because the differences in etiology, management and outcome are significant, accurate and timely diagnosis is imperative. OBJECTIVE: The purpose of this report is to present our experience with embolotherapy for treatment of high-flow priapism in three children. PATIENTS AND METHODS: This was a retrospective study. During an 18-month period, three boys ranging in age from 6 to 15 years presented with priapism. All three children were treated with embolotherapy. RESULTS: All three children were successfully treated with angiography and embolotherapy. One boy had a presentation that initially raised the possibility of low-flow priapism. No complications occurred, and to date all children are able to maintain normal erections. CONCLUSION: Subselective transcatheter embolization is the procedure of choice for high-flow priapism. In cases where priapism persists despite adequate therapy, angiography might be useful to exclude high-flow disease. In children with high-flow priapism, selective occlusion of the penile arteriovenous fistula led to detumescence and normal erectile function. PMID- 17415603 TI - Bilhemia: a fatal complication following percutaneous placement of a transhepatic inferior vena cava catheter in a child. AB - A transhepatic central venous catheter was implanted in a 2-year-old child with a history of multiple venous access procedures and superior and inferior vena cava thrombosis. After 2 weeks, inadvertent dislodgement of the catheter was complicated by a biloma. The biloma was percutaneously drained, but a biliary venous fistula led to a rapidly progressive and fatal bilhemia. We report this case as an infrequent complication of transhepatic catheterization. PMID- 17415604 TI - Magnetic resonance angiography of an ipsilateral double aortic arch due to persistent left fourth and fifth aortic arches. AB - Persistence of a fifth aortic arch with an ipsilateral fourth arch manifested as a double-lumen aortic arch (DLAA) is a rare congenital anomaly. We present one of the first cases diagnosed by magnetic resonance angiography (MRA) in a 7-year-old girl referred for treatment of subaortic membrane and aortic regurgitation. PMID- 17415608 TI - Methionine aminopeptidase II: A molecular chaperone for sarcoplasmic reticulum calcium ATPase. AB - The monoclonal antibody to the beta-subunit of H(+)/K(+)-ATPase (mAbHKbeta) cross reacts with a protein that acts as a molecular chaperone for the structural maturation of sarcoplasmic reticulum (SR) Ca(2+)-ATPase. We partially purified a mAbHKbeta-reactive 65-kDa protein from Xenopus ovary. After in-gel digestion and peptide sequencing, the 65-kDa protein was identified as methionine aminopeptidase II (MetAP2). The effects of MetAP2 on SR Ca(2+)-ATPase expression were examined by injecting the cRNA for MetAP2 into Xenopus oocytes. Immunoprecipitation and pulse-chase experiments showed that MetAP2 was transiently associated with the nascent SR Ca(2+)-ATPase. Synthesis of functional SR Ca(2+)-ATPase was facilitated by MetAP2 and prevented by injecting an antibody specific for MetAP2. These results suggest that MetAP2 acts as a molecular chaperone for SR Ca(2+)-ATPase synthesis. PMID- 17415609 TI - Modulation of the permeability transition pore by inhibition of the mitochondrial K(ATP) channel in liver vs. brain mitochondria. AB - Inhibition of the mitochondrial K(ATP) (mitoK(ATP)) channel abrogates the beneficial effects of preconditioning induced by a brief episode of sublethal ischemia. We studied the effect of 5-hydroxydecanoate, a well-known inhibitor of the mitoK(ATP) channel, on swelling of isolated liver and brain mitochondria. Volume changes were determined by measurement of light absorbance at 540 nm. Mitochondrial swelling induced by adding Ca(2+ )ions correlated with opening of the permeability transition pore as shown by modulation by 1 microM cyclosporin A. In brain mitochondria, 5-hydroxydecanoate did not significantly affect Ca(2+) induced swelling. In contrast, 50 or 500 microM 5-hydroxydecanoate increased swelling of liver mitochondria by 9.7 +/- 5.1% (n = 6, P = 0.057) and 29.4 +/- 1.4% (n = 5, P < 0.0001), respectively. The effect of 5-hydroxydecanoate was blocked by cyclosporin A and was dependent on the presence of potassium in the medium. In medium containing 200 microM ATP to inhibit the mitoK(ATP )channel, 5 hydroxydecanoate did not further increase Ca(2+)-induced swelling. We conclude that inhibition of the mitoK(ATP) channel exerts its detrimental effect by facilitation of permeability transition pore opening. PMID- 17415610 TI - Developmental expression of the outer hair cell motor prestin in the mouse. AB - The development of motor protein activity in the lateral membrane of the mouse outer hair cell (OHC) from postnatal day 5 (P5) to P18 was investigated under whole-cell voltage clamp. Voltage-dependent, nonlinear capacitance (C (v)), which represents the conformational fluctuations of the motor molecule, progressively increased during development. At P12, the onset of hearing in the mouse, C (v) was about 70% of the mature level. C (v) saturated at P18 when hearing shows full maturation. On the other hand, C (lin), which represents the membrane area of the OHC, showed a relatively small increase with development, reaching steady state at P10. This early maturation of linear capacitance is further supported by morphological estimates of surface area during development. These results, in light of recent prestin knockout experiments and our results with quantitative polymerase chain reaction, suggest that, rather than the incorporation of new motors into the lateral membrane after P10, molecular motors mature to augment nonlinear capacitance. Thus, current estimates of motor protein density based on charge movement may be exaggerated. A corresponding indicator of motor maturation, the motor's operating voltage midpoint, V (pkcm), tended to shift to depolarized potentials during postnatal development, although it was unstable prior to P10. However, after P14, V (pkcm) reached a steady-state level near -67 mV, suggesting that intrinsic membrane tension or intracellular chloride, each of which can modulate V (pkcm), may mature at P14. These developmental data significantly alter our understanding of the cellular mechanisms that control cochlear amplification and provide a foundation for future analysis of genetic modifications of mouse auditory development. PMID- 17415611 TI - Microbial environmental genomics. PMID- 17415612 TI - Across space and time: social responses to large-scale biophysical systems. AB - The conceptual rubric of ecosystem management has been widely discussed and deliberated in conservation biology, environmental policy, and land/resource management. In this paper, I argue that two critical aspects of the ecosystem management concept require greater attention in policy and practice. First, although emphasis has been placed on the "space" of systems, the "time" -- or rates of change -- associated with biophysical and social systems has received much less consideration. Second, discussions of ecosystem management have often neglected the temporal disconnects between changes in biophysical systems and the response of social systems to management issues and challenges. The empirical basis of these points is a case study of the "Crown of the Continent Ecosystem," an international transboundary area of the Rocky Mountains that surrounds Glacier National Park (USA) and Waterton Lakes National Park (Canada). This project assessed the experiences and perspectives of 1) middle- and upper-level government managers responsible for interjurisdictional cooperation, and 2) environmental nongovernment organizations with an international focus. I identify and describe 10 key challenges to increasing the extent and intensity of transboundary cooperation in land/resource management policy and practice. These issues are discussed in terms of their political, institutional, cultural, information-based, and perceptual elements. Analytic techniques include a combination of environmental history, semistructured interviews with 48 actors, and text analysis in a systematic qualitative framework. The central conclusion of this work is that the rates of response of human social systems must be better integrated with the rates of ecological change. This challenge is equal to or greater than the well-recognized need to adapt the spatial scale of human institutions to large-scale ecosystem processes and transboundary wildlife. PMID- 17415614 TI - The dynamic of the t-haplotype in wild populations of the house mouse Mus musculus domesticus in Israel. AB - The t-haplotype, a variant of the proximal part of the mouse chromosome 17, is composed of at least four inversions and is inherited as a single genetic unit. The haplotype causes embryonic mortality or male sterility when homozygous. Genes within the complex are responsible for distortion of Mendelian transmission ratio in males. Thus, the t-haplotype in heterozygous males is transferred to over 95% of the progeny. We examined the dynamic and behavior of the t-haplotype in wild populations of the house mouse in Israel. The Israeli populations show high frequency (15%-20%) of both partial and complete t-carrying mice, supporting the suggestion that the t-complex evolved in the M. domesticus line in the Israeli region. In one population that had the highest frequency of t-carrying individuals, we compared the level of gene diversity between t-carrying and normal mice in the marker's loci: H-2 locus of the major histocompatibility complex (MHC) on the t-haplotype of chromosome 17, three microsatellites on other chromosomes, and the mitochondrial D-loop. Genetic variability was high in all tested loci in both t and (+) mice. All t mice carried the same chromosome and showed the same H-2 haplotype. While t-carrying mice showed significant H-2 heterozygotes access, (+) mice expressed significant H-2 heterozygote deficiency. There were no differences in the level of gene diversity between t and (+) mice in the other loci. Heterozygosity level at the MHC may be an additional factor in the selective forces balancing the t-haplotype polymorphism. PMID- 17415615 TI - Atypical lung changes in a 19-year-old woman with Crohn's disease. PMID- 17415616 TI - Chemical organisation theory. AB - Complex dynamical reaction networks consisting of many components that interact and produce each other are difficult to understand, especially, when new component types may appear and present component types may vanish completely. Inspired by Fontana and Buss (Bull. Math. Biol., 56, 1-64) we outline a theory to deal with such systems. The theory consists of two parts. The first part introduces the concept of a chemical organisation as a closed and self maintaining set of components. This concept allows to map a complex (reaction) network to the set of organisations, providing a new view on the system's structure. The second part connects dynamics with the set of organisations, which allows to map a movement of the system in state space to a movement in the set of organisations. The relevancy of our theory is underlined by a theorem that says that given a differential equation describing the chemical dynamics of the network, then every stationary state is an instance of an organisation. For demonstration, the theory is applied to a small model of HIV-immune system interaction by Wodarz and Nowak (Proc. Natl. Acad. USA, 96, 14464-14469) and to a large model of the sugar metabolism of E. Coli by Puchalka and Kierzek (Biophys. J., 86, 1357-1372). In both cases organisations where uncovered, which could be related to functions. PMID- 17415617 TI - Weight loss maintenance in African-American women: focus group results and questionnaire development. AB - BACKGROUND: African-American women are disproportionately affected by obesity. Weight loss can occur, but maintenance is rare. Little is known about weight loss maintenance in African-American women. OBJECTIVES: (1) To increase understanding of weight loss maintenance in African-American women; (2) to use the elicitation procedure from the theory of planned behavior (TPB) to define the constructs of attitude, subjective norms, and perceived behavioral control regarding weight loss and maintenance; and (3) to help develop a relevant questionnaire that can be used to explore weight loss and maintenance in a large sample of African Americans. DESIGN: Seven focus groups were conducted with African-American women: four with women successful at weight loss maintenance, three with women who lost weight but regained it. Discussions centered on weight loss and maintenance experiences. PARTICIPANTS: Thirty-seven African-American women. APPROACH: Content analysis of focus group transcripts. RESULTS: Weight loss maintainers lost 22% of body weight. They view positive support from others and active opposition to cultural norms as critical for maintenance. They struggle with weight regain, but have strategies in place to lose weight again. Some maintainers struggle with being perceived as sick or too thin at their new weight. Regainers and maintainers struggle with hairstyle management during exercise. The theoretical constructs from TPB were defined and supported by focus group content. CONCLUSIONS: A weight loss questionnaire for African Americans should include questions regarding social support in weight maintenance, the importance of hair management during exercise, the influence of cultural norms on weight and food consumption, and concerns about being perceived as too thin or sick when weight is lost. PMID- 17415618 TI - Value-based insurance design: a "clinically sensitive, fiscally responsible" approach to mitigate the adverse clinical effects of high-deductible consumer directed health plans. PMID- 17415619 TI - Homeless people's perceptions of welcomeness and unwelcomeness in healthcare encounters. AB - BACKGROUND: Homeless people face many barriers to obtaining health care, and their attitudes toward seeking health care services may be shaped in part by previous encounters with health care providers. OBJECTIVE: To examine how homeless persons experienced "welcomeness" and "unwelcomeness" in past encounters with health care providers and to characterize their perceptions of these interactions. DESIGN: Qualitative content analysis of 17 in-depth interviews. PARTICIPANTS: Seventeen homeless men and women, aged 29-62 years, residing at 5 shelters in Toronto, Canada. APPROACH: Interpretive content analysis was performed using iterative stages of inductive coding. Interview transcripts were analyzed using Buber's philosophical conceptualization of ways of relating as "I It" (the way persons relate to objects) and "I-You" (the way persons relate to dynamic beings). RESULTS: Most participants perceived their experiences of unwelcomeness as acts of discrimination. Homelessness and low social class were most commonly cited as the perceived basis for discriminatory treatment. Many participants reported intense emotional responses to unwelcoming experiences, which negatively influenced their desire to seek health care in the future. Participants' descriptions of unwelcoming health care encounters were consistent with "I-It" ways of relating in that they felt dehumanized, not listened to, or disempowered. Welcoming experiences were consistent with "I-You" ways of relating, in that patients felt valued as a person, truly listened to, or empowered. CONCLUSIONS: Homeless people's perceptions of welcomeness and unwelcomeness are an important aspect of their encounters with health care providers. Buber's "I-It" and "I-You" concepts are potentially useful aids to health care providers who wish to understand how welcoming and unwelcoming interactions are fostered. PMID- 17415620 TI - Care management dosage. AB - BACKGROUND: The care of patients with complex illnesses requires careful management, but systems of care management (CM) vary in their structure and effectiveness. OBJECTIVE: To create a framework identifying components of broad based CM interventions and validate the framework, including using this framework to evaluate the contribution of varying components on outcomes of patients with chronic illness. DESIGN: We create the framework using retrospective information about CM activities and services over 12 months and categorize it using cluster and factor analysis. We then validate this framework through content and criterion techniques. Content validity is assessed through a Delphi study and criterion validity through relationship of the dosage measures and patterns of care to process and outcomes measures. PARTICIPANTS: Patients with diabetes and/or cardiovascular disease receiving CM services in a model known as Care Management Plus implemented in primary care. RESULTS: Six factors of CM activity were identified, including a single dosage summary measure and 5 separate patterns of care. Of these, the overall dosage summary measure, face-to-face time, duration of follow-up, and breadth of services were all related to improved processes for hemoglobin A1c and LDL testing and control. Brief intense patterns of care and high face-to-face care manager time were also related to improved outcomes. CONCLUSIONS: Using this framework, we isolate components of a CM intervention directly related to improved process of care or patient outcomes. Current efforts to structure CM to include face-to-face time and multiple diseases are discussed. PMID- 17415621 TI - A study on the social structure and dispersal patterns of hamadryas baboons living in a commensal group at Taif, Saudi Arabia. AB - Three levels of hamadryas social structure--the one male unit (OMU), the band, and the troop--have been observed at all sites studied, but a fourth--the clan- has been observed at only one site, Erer-Gota, Ethiopia, during a longitudinal check of the dispersion of identified individuals. The clan is important since it appears to provide the basis for male philopatry, although comparative data is needed from other sites to confirm this. We studied a huge commensal group of hamadryas baboons (over 600 animals) in Saudi Arabia. We put ear tags on baboons between 1998 and 2004 and analyzed social structure, relying on the interactions of these tagged animals by focusing especially on their dispersal patterns from OMUs. OMU membership tended to be looser than that of the Ethiopian hamadryas. Females tended to shift between OMUs on an individual basis in our study group, whereas the collapse of an OMU was a major occasion of adult female transfer in Ethiopia. We found neither stable bands (a "band" in our study group was defined as a regional assemblage of OMUs) nor clans that lasted for several years. Some OMUs moved and transferred into neighboring areas over both the short and long term. Further, some post-adolescent males appeared to move out of the study area. The ratio of adult females in an OMU in our study group was larger than for any other documented study site, and this may be the reason for enhanced female transfer between OMUs. A large proportion of the adolescent females showed no clear membership to OMUs, and no "initial units" (commonly observed in Ethiopia) were discernible. The ease with which young males acquired adult females at the study site must have disrupted the formation of a clan, a "male-bonded society." PMID- 17415622 TI - Functional characterization of two distinct Mg(2+) extrusion mechanisms in cardiac sarcolemmal vesicles. AB - Cardiac ventricular myocytes extrude a sizeable amount of their total Mg(2+) content upon stimulation by beta-adrenergic agonists. This extrusion occurs within a few minutes from the application of the agonist, suggesting the operation of rapid and abundantly represented Mg(2+) transport mechanisms in the cardiac sarcolemma. The present study was aimed at characterizing the operation of these transport mechanisms under well defined conditions. Male Sprague-Dawley rats were used to purify a biochemical standardized preparation of sealed rat cardiac sarcolemmal vesicles. This experimental model has the advantage that trans-sarcolemmal cation transport can be studied under specific extra- and intra vesicular ionic conditions, in the absence of intracellular organelles, and buffering or signaling components. Magnesium ion (Mg(2+)) transport was assessed by atomic absorbance spectrophotometry. The results reported here indicate that: (1) sarcolemma vesicles retained trapped intravesicular Mg(2+) in the absence of extravesicular counter-ions; (2) the addition of Na(+) or Ca(2+) induced a rapid and concentration-dependent Mg(2+) extrusion from the vesicles; (3) co-addition of maximal concentrations of Na(+) and Ca(2+) resulted in an additive Mg(2+) extrusion; (4) Mg(2+ )extrusion was blocked by addition of amiloride or imipramine; (5) pre-treatment of sarcolemma vesicles with alkaline phosphatase at the time of preparation completely abolished Na(+)- but not Ca(2+)-induced Mg(2+) extrusion; (6) Na(+)-dependent Mg(2+) transport could be restored by stimulating vesicles loaded with protein kinase A catalytic subunit and ATP with membrane permeant cyclic-AMP analog; (7) extra-vesicular Mg(2+) could be accumulated in exchange for intravesicular Na(+) via a mechanism inhibited by amiloride or alkaline phosphatase treatment; (8) Mg(2+) accumulation could be restored via cAMP/protein kinase A protocol. Overall, these data provide compelling evidence for the operation of distinct Na(+)- and Ca(2+)-dependent Mg(2+) extrusion mechanisms in sarcolemma vesicles. The Na(+)-dependent mechanism appears to be specifically activated via protein kinase A/cAMP-dependent phosphorylation process, and can operate in either direction based upon the cation concentration gradient across the sarcolemma. The Ca(2+)-dependent mechanism, instead, only mediates Mg(2+) extrusion in a cAMP-independent manner. PMID- 17415623 TI - Regulation of skeletal muscle differentiation in fibroblasts by exogenous MyoD gene in vitro and in vivo. AB - MyoD of the myogenic regulatory factors (MRFs) family regulates the skeletal muscle differentiation program. In this study, stably transfected NIH3T3-derived cell lines were established, in which exogenous MyoD was expressed at high levels. Transcriptional activation of endogenous muscle regulatory gene and induction towards the skeletal muscle lineages were observed with phase-contrast microscopy when continuously cultured in vitro. Moreover, to determine their ability of myogenic formation in vivo, the transfected cells were implanted in nude mice subcutaneously for up to 10 weeks. The morphological characterization of inductive cells was observed using transmission electron microscope and histological staining. Myogenesis of fibroblasts incubated in the medium was activated by overexpression of MyoD, and the cells were accumulated and fused into multinucleated myotubes. Correlatively, RT-PCR and immunohistochemistry confirmed the increased expression of characteristic downstream molecule myogenin and mysion heavy chains during myogenic differentiation. Ecoptic myogenesis was found and remained stable phenotype when the transfected cells were seeded in vivo. Our results suggest that MyoD can be considered to be a determining factor of myogenic lineages, and it may play an important role in the cell therapy and cell-mediated gene therapy of the skeletal muscle. PMID- 17415624 TI - Rice allelopathy induced by methyl jasmonate and methyl salicylate. AB - Methyl jasmonate (MeJA) and methyl salicylate (MeSA) are important signaling molecules that induce plant defense against insect herbivores and microbial pathogens. We tested the hypothesis that allelopathy is an inducible defense mechanism, and that the JA and SA signaling pathways may activate allelochemicals release. Exogenous application of MeJA and MeSA to rice (Oryza sativa L.) enhanced rice allelopathic potential and led to accumulation of phenolics, an increase in enzymatic activities, and gene transcription of phenylalanine ammonia lyase (PAL) and cinnamate 4-hydroxylase (C4H), two key enzymes in the phenylpropanoid pathway. Aqueous extracts of the leaves of rice IAC165, a putative allelopathic variety, treated with MeSA (5 mM) or MeJA (0.05 mM), showed increased inhibitory effects (25 and 21%, respectively) on root growth of barnyardgrass (Echinochloa crus-galli L.), and increased inhibitory effects (18 and 23%, respectively) on shoot growth. Aqueous extracts from leaves of Huajingxian 1 rice, a putative nonallelopathic variety treated with MeJA and MeSA, caused 63 and 24% inhibition of root growth in barnyardgrass seedlings. The root exudates of both IAC165 and Huajingxian 1 plants treated with MeJA and MeSA for 48 hr also showed significant increases in their inhibitory effects on root growth of barnyardgrass seedlings. At the four-leaf stage, levels of 3,4 hydroxybenzoic acid, vanillic acid, coumaric acid, and ferulic acid that accumulated in the leaves were 5.3-, 31.3-, 2.2-, and 1.7-fold higher in response to MeJA exposure, and 3.3-, 13.1-, 2.0-, and 2.2-fold higher in response to MeSA. Treatments of MeSA and MeJA enhanced the PAL activity in the rice leaves up to 52.3 and 80.1%, respectively, whereas C4H activity was increased by 40.2 and 67%. Gene transcription of PAL and C4H in rice leaves significantly increased after the plants were subjected to treatment with MeJA and MeSA. These results suggest that allelopathy may be an active defense mechanism, and that plant signaling compounds are potentially valuable in its regulation. PMID- 17415625 TI - Induction of plant volatiles by herbivores with different feeding habits and the effects of induced defenses on host-plant selection by thrips. AB - Induced plant responses to attack by chewing insects have been intensively studied, but little is known about plant responses to nonchewing insects or to attack by multiple herbivores with different feeding habits. We examined volatile emissions by tobacco, Nicotiana tabacum, in response to feeding by the piercing sucking insect western flower thrips (WFT), Frankliniella occidentalis, the chewing herbivore Heliothis virescens, and both herbivores simultaneously. In addition, we examined the effects of herbivore-induced plant defenses on host plant selection by WFT. Plants responded to thrips feeding by consistently releasing five compounds. Simultaneous feeding by WFT and H. virescens elicited the same 11 compounds emitted in response to caterpillar feeding alone; however, two compounds, alpha-humulene and caryophyllene oxide, were produced in greater amounts in response to simultaneous herbivory. In choice tests, thrips consistently preferred uninduced plants over all other treatments and preferred plants damaged by caterpillars and those treated with caterpillar saliva over those treated with caterpillar regurgitant. The results are consistent with a previous finding that caterpillar regurgitant induces the release of significantly more volatile nicotine than plants damaged by caterpillars or plants treated with caterpillar saliva. A repellent effect of nicotine on WFT was confirmed by encircling unwounded plants with septa releasing volatile nicotine. Our results provide the first direct evidence that thrips feeding induces volatile responses and indicates that simultaneous herbivory by insects with different feeding habits can alter volatile emissions. In addition, the findings demonstrate that induced plant responses influence host-plant selection by WFT and suggest that the induction of volatile nicotine may play a role in this process. PMID- 17415626 TI - Foliar phenolics are differently associated with Epirrita autumnata growth and immunocompetence. AB - The quality of available food may affect insect herbivores directly (via growth and survivorship) and/or indirectly (by modifying insect vulnerability to parasitoids and pathogens). We examined the relationship between different phenolic compounds, belonging to various phenolic groups, in Betula pubescens spp. czerepanovii (mountain birch) foliage and the larval performance of the geometrid Epirrita autumnata (autumnal moth). Direct effects on insect performance were described by pupal weight, developmental rate, and survivorship; indirect effects were described by the encapsulation rate of an implant inserted into the insect hemocoel, a commonly used way to describe insect immune defense. We found profound differences in the effects of different phenolic categories: several individual hydrolyzable tannins were associated positively with larval performance but negatively with level of immune defense, whereas flavonoid glycosides were inversely related to larval survival but showed no association with the larvae immune defense. PMID- 17415627 TI - Mapping and ablation: a worldwide perspective. AB - Currently, many ablative techniques have reported excellent long-term results in restoring and maintaining sinus rhythm among patients with paroxysmal and persistent AF. Catheter ablation of AF should not be performed too late over time when recurrent paroxysmal AF progresses to the persistent or to the permanent form. Among patients with permanent AF, the stepwise approach requires very extensive lesions in both the left and right atrium to obtain the same success rate as reported by CPVA as performed in Milan many years ago. Long-term prospective multicenter randomized studies comparing the impact of medical therapy with catheter ablation strategy on "hard" outcomes such as morbidity and mortality are required to better define the patient population that may mostly benefit from ablation at the lowest risk and acceptable cost. PMID- 17415628 TI - Fighting against sudden death: a single or multidisciplinary approach. AB - There are many causes of sudden death ranging from accidents and suicide to vascular events and arrhythmias. Most sudden deaths will occur in people who have not been diagnosed with a serious heart condition but at a very low annual rate. Many of these events are probably vascular and might be prevented by reducing the risk of developing coronary disease. Only a minority of sudden deaths occur in people with established cardiac disease, but in patients with major structural heart disease, the annual rate is high. The causes of sudden death are many in this clinical setting also, but dominated by ventricular arrhythmias and vascular events. There is good evidence that conventional treatments for heart failure, including ACE inhibitors, beta-blockers, aldosterone antagonists and cardiac resynchronization devices reduce the risk of sudden death. Evidence that statins, aspirin or revascularisation are safe or effective in patients with heart failure is currently lacking. Implantable defibrillators confer a small but definite additional survival advantage by treating arrhythmias that have not been prevented. PMID- 17415629 TI - Variability v.s. synchronicity of neuronal activity in local cortical network models with different wiring topologies. AB - Dynamical behavior of a biological neuronal network depends significantly on the spatial pattern of synaptic connections among neurons. While neuronal network dynamics has extensively been studied with simple wiring patterns, such as all-to all or random synaptic connections, not much is known about the activity of networks with more complicated wiring topologies. Here, we examined how different wiring topologies may influence the response properties of neuronal networks, paying attention to irregular spike firing, which is known as a characteristic of in vivo cortical neurons, and spike synchronicity. We constructed a recurrent network model of realistic neurons and systematically rewired the recurrent synapses to change the network topology, from a localized regular and a "small world" network topology to a distributed random network topology. Regular and small-world wiring patterns greatly increased the irregularity or the coefficient of variation (Cv) of output spike trains, whereas such an increase was small in random connectivity patterns. For given strength of recurrent synapses, the firing irregularity exhibited monotonous decreases from the regular to the random network topology. By contrast, the spike coherence between an arbitrary neuron pair exhibited a non-monotonous dependence on the topological wiring pattern. More precisely, the wiring pattern to maximize the spike coherence varied with the strength of recurrent synapses. In a certain range of the synaptic strength, the spike coherence was maximal in the small-world network topology, and the long range connections introduced in this wiring changed the dependence of spike synchrony on the synaptic strength moderately. However, the effects of this network topology were not really special in other properties of network activity. PMID- 17415630 TI - Tactile perception in adults with autism: a multidimensional psychophysical study. AB - Although sensory problems, including unusual tactile sensitivity, are heavily associated with autism, there is a dearth of rigorous psychophysical research. We compared tactile sensation in adults with autism to controls on the palm and forearm, the latter innervated by low-threshold unmyelinated afferents subserving a social/affiliative submodality of somatosensation. At both sites, the groups displayed similar thresholds for detecting light touch and innocuous sensations of warmth and cool, and provided similar hedonic ratings of the pleasantness of textures. In contrast, increased sensitivity to vibration was seen in the autism group on the forearm, along with increased sensitivity to thermal pain at both sites. These findings suggest normal perception along with certain areas of enhanced perception in autism, consistent with previous studies. PMID- 17415631 TI - Interferon-gamma promotes differentiation of neural progenitor cells via the JNK pathway. AB - It has been reported that interferon-gamma (IFN-gamma) facilitates differentiation of PC-12 cells and murine adult neural stem cells. Here we show that IFN-gamma promotes the differentiation of C17.2 neural progenitor cells (NPC) into a neuronal phenotype characterized by neurite outgrowth and the expression of the neuronal marker protein beta-III tubulin. IFN-gamma induced an increase in the activity c-jun N-terminal kinase (JNK) without affecting activities of extracellular signal-regulated kinases (ERKs 1 and 2). An inhibitor of JNK blocked the ability of IFN-gamma to promote differentiation of NPC into neurons, whereas an inhibitor of ERKs 1 and 2 did not. Our findings show that the pro-inflammatory cytokine, IFN-gamma has the potential to stimulate neurogenesis, suggesting roles for this cytokine in development and repair of the nervous system. PMID- 17415632 TI - A meta-analysis of antibiotic therapy for active ulcerative colitis. AB - To systematically evaluate the efficacy of antibacterial therapy in ulcerative colitis, we carried out a meta-analysis of controlled clinical trials. Within the time period 1966 through September 2006, PUBMED, EMBASE, and SCOPUS were searched for clinical trial studies that investigated the efficacy of antibiotics in ulcerative colitis. We considered clinical remission as our key outcome of interest. Of 122 studies, 10 randomized placebo-controlled clinical trials matched our criteria and were included in the analysis (530 patients). All the studies used antibiotics as an adjunct therapy to conventional treatment of ulcerative colitis (i.e., corticosteroids and 5-aminosalycilic acid). Pooling of these trials yielded odds ratio (OR) of 2.14 (95% confidence interval [CI], 1.48 3.09; P<0.0001) in favor of antimicrobial therapy. Meta-analysis of short-term trials (5-14 days) showed a higher rate of clinical remission in patients treated with antibiotics (OR, 2.02; 95% CI, 1.36-3). These results suggest that adjunctive antibacterial therapy is effective for induction of clinical remission in ulcerative colitis. PMID- 17415633 TI - Bone morphogenetic protein 7 is elevated in patients with chronic liver disease and exerts fibrogenic effects on human hepatic stellate cells. AB - Hepatic stellate cells (HSCs) are the main extracellular matrix (ECM)-producing cells in liver fibrogenesis. The excessive synthesis of ECM proteins deteriorates hepatic architecture and results in liver fibrosis and cirrhosis. This study investigated the role of bone morphogenetic protein 7 (BMP7) as a member of the transforming growth factor (TGF)-beta superfamily in chronic liver disease. Plasma levels of BMP7 were significantly elevated in patients with chronic liver disease compared with healthy controls. Immunohistochemistry of cirrhotic human liver demonstrated upregulated BMP7 protein expression in hepatocytes as compared with normal human liver. Because gene expression for all putative BMP7 receptors was induced during the culture activation process of primary human HSCs, we studied the effects of BMP7 on hTERT immortalized human HSCs in vitro. BMP7, as expressed and secreted after infection with adenoviruses encoding BMP7 (AdBMP7), increased proliferation of HSCs. The mRNA and protein expression of type I collagen and fibronectin was increased in BMP7-stimulated HSCs. Elevated systemic and hepatic levels of BMP7 in patients with chronic liver disease may contribute to progression of liver fibrogenesis in vivo. PMID- 17415634 TI - Da-Cheng-Qi-Tang promotes the recovery of gastrointestinal motility after abdominal surgery in humans. AB - In order to examine the effects of Da-Cheng-Qi-Tang (DCQT) on gastrointestinal motility functions after abdominal surgery in humans, 33 patients with abdominal surgeries and 36 patients with cholecystectomies were divided into the DCQT and the control groups at random. Electrogastrography (EGG) and gastroduodenojejunal manometry was performed and the levels of plasma motilin were measured by radioimmunoassay. The results were as follows: (1) on the day of surgery, the ratio of EGG normal frequency in the DCQT group was higher than in the control group (P=0.0016); (2) the power of EGG in the DCQT group was higher than in the control group on the second and third days after surgeries (P=0.0011 and P=0.0215, respectively); (3) the percentage of normal bowel peristalsis was significantly higher in the DCQT group than in the control group (P<0.01); and (4) in the DCQT group, the plasma motilin level reached its peak earlier than in the control group. Our results suggest that DCQT can increase plasma motilin, enhance gastrointestinal motility, improve gastric dysrythmia, and reduce gastroparesis after abdominal surgery. PMID- 17415635 TI - Further validation of the IBS-QOL: female Mexican IBS patients have poorer quality of life than females from North Carolina. AB - The Irritable Bowel Syndrome (IBS)-Quality of Life (QOL) is the most extensively validated health-related quality of life (HRQOL)-specific instrument for IBS with appropriate evidence for psychometric validity. Our aim was to linguistically validate the IBS-QOL for Mexico according to standard guidelines, to conduct further psychometric validation, and to compare the HRQOL between IBS patients from Mexico and North Carolina (University of North Carolina). Construct validity was tested by correlating scores from the Mexican Spanish IBS-QOL with those for anxiety and depression obtained by the Hospital Anxiety and Depression scale. Also, HRQOL from Rome I female IBS patients who consulted a tertiary referral center in Mexico was compared with that of female patients from UNC matched by age and bowel habit. A general univariate linear model was done to determine the most important variable over HRQOL, place of origin, or bowel habit. The majority of the IBS-QOL items had a negative correlation with depression as well as with anxiety. Compared to patients from UNC, the Mexican ones reported significant lower scores on Body Image and Health Worry and a trend in Interference with activities and in the Overall score. There were some differences in Dysphoria and Interference that were related to bowel habit, independently of the place of origin. In conclusion, the IBS-QOL validated in Mexican Spanish has shown construct validity. Using this instrument we found that female IBS patients who consulted a tertiary referral center in Mexico have lower HRQOL than those in North Carolina at least in factors such as Body Image and Health Worry. PMID- 17415636 TI - Positive CAGE screen correlates with cirrhosis in veterans with chronic hepatitis C. AB - The current study examines the relationship between problem alcohol use and severity of liver disease by self-administered questionnaires using both the CAGE questionnaire and beverage-specific quantity-frequency questions. The cohort consisted of 38 patients with cirrhosis (10 with decompensated liver disease) and 62 with mild fibrosis (stage 0-1), of comparable mean age and estimated duration of infection. Although mean alcohol consumption was similar in both groups, a positive CAGE screen (defined as two or more affirmative answers) was significantly more common among cirrhotics (OR = 5.24; 95% CI, 1.78-15.39) and tended to be associated with decompensated liver disease (OR = 13.3; 95% CI, 0.67 256) among cirrhotics. In multivariate analysis, only inflammatory grade on liver biopsy (OR = 67.7; 95% CI, 10.6-431) and positive CAGE score (OR = 8.09; 95% CI, 1.15-57.1) were independent predictors of cirrhosis. These findings suggest that the CAGE questionnaire predicts advanced liver disease better than daily or lifetime drinking measures. PMID- 17415637 TI - Oral erythromycin accelerates impaired gastrointestinal motility after endoscopic mucosal resection. AB - Gastrointestinal motility may be impaired after endoscopic mucosal resection of gastric lesions. We investigated whether oral erythromycin could improve motility. Twenty patients were divided randomly into groups that received oral omeprazole with or without erythromycin. Motility was recorded overnight at 3 days before and 4 days after endoscopic resection using a microtransducer probe. In the group without erythromycin, gastric phase III activity decreased significantly after endoscopic resection, while it was increased significantly by erythromycin (P < 0.01). After resection, there were significantly more gastric phase III events in the erythromycin group (P < 0.05). The interval between the start of the evening meal and the initial gastric phase III activity was significantly prolonged after resection, while this interval was significantly shortened by erythromycin (P < 0.05). The gastric phase III cycle length was also significantly shortened by erythromycin (P < 0.05). Postprandial and fasting gastrointestinal motility were impaired after endoscopic resection, and postprandial as well as fasting motility were improved by oral erythromycin. PMID- 17415638 TI - Poorly differentiated adenocarcinoma arising from a hyperplastic polyp: which polyps should be endoscopically resected? PMID- 17415639 TI - Melanoma of the anus disguised as hemorrhoids: surgical management illustrated by a case report. PMID- 17415641 TI - Detection and treatment of early flat and depressed colorectal cancer using high magnification chromoscopic colonoscopy: a change in paradigm for Western endoscopists? AB - Japanese researchers reported flat and depressed colorectal lesions in the 1980s. Such lesions were thought irrelevant to Western populations and described as "Phantom" or "Akitas" carcinoma. Many depressed neoplasms arise through the de novo pathogenic sequence and demonstrate early invasive characteristics. All investigators report difficulties in identifying flat and depressed lesions using conventional colonoscopy. Failure to detect and treat such lesions may be responsible for the current shortfalls in secondary colorectal cancer prevention. Given the introduction of colorectal cancer screening programs in the United Kingdom, Europe, and the United States, it is essential to re-evaluate the significance of flat lesions as applicable to Western cohorts and explore the safety and efficacy of new endoscopic technology and interventional therapeutics. PMID- 17415640 TI - Zinc and the liver: an active interaction. AB - Zinc is an essential trace element, exerting important antioxidant, anti inflammatory, and antiapoptotic effects. It affects growth and development and participates in processes such as aging and cancer induction. The liver is important for the regulation of zinc homeostasis, while zinc is necessary for proper liver function. Decreased zinc levels have been implicated in both acute and chronic liver disease states, and zinc deficiency has been implicated in the pathogenesis of liver diseases. Zinc supplementation offers protection in experimental animal models of acute and chronic liver injury, but these hepatoprotective properties have not been fully elucidated. In the present review, data on zinc homeostasis, its implication in the pathogenesis of liver diseases, and its effect on acute and chronic liver diseases are presented. It is concluded that zinc could protect against liver diseases, although up to now the underlying pathophysiology of zinc and liver interactions have not been defined. PMID- 17415642 TI - Computed tomography of pancreatic implantation with malignant transformation of leiomyomatosis peritonealis disseminata in a man. AB - Leiomyomatosis peritonealis disseminata (LPD) is a rare condition in females. We report a case of pancreatic implantation with malignant transformation of LPD in a man. A 42-year-old man presented with a 16-month history of persistent pain in the right upper abdomen with dorsolumbar radiating pain and weight loss. Spiral computed tomography revealed diffuse enlargement of pancreas with multiple, miliary, micronodular, low-density lesions. To the best of our knowledge, it is the first report of the pancreatic implantation of LPD and the third report of LPD in a male. PMID- 17415643 TI - Combination of adrenaline injection and detachable snare application as haemostatic preventive measures, before polypectomy of large colonic polyps in children. PMID- 17415644 TI - Chronic hypergastrinemia: causes and consequences. AB - The hormone gastrin plays 2 important roles in gastrointestinal physiology--1 as a major factor in meal-stimulated gastric acid secretion and the other as a trophic hormone for epithelial and enterochromaffin cells. These roles are exaggerated to the point of pathology under conditions of chronic hypergastrinemia as exemplified by the Zollinger-Ellison syndrome and pernicious anemia. More recently, the concern about the potential risk of chronic hypergastrinemia has risen because of the widespread use of proton pump inhibitors for maintenance therapy in reflux esophagitis. For this reason, we present a concise overview of the origin, causes, and potential risks of chronic hypergastrinemia. PMID- 17415645 TI - A fulminant enterocolitis. PMID- 17415646 TI - Gut toxicity during hemopoietic stem cell transplantation may predict acute graft versus-host disease severity in patients. AB - Graft-versus-host disease (GVHD) is the primary complication of allogeneic, hemopoietic, stem cell transplantation (HSCT). Murine models suggest that gut toxicity, induced by the intensive chemotherapy preceding hematopoietic stem cell infusion, aggravates systemic GVHD. In HSCT patients gut toxicity correlates with chemotherapy intensity. The present study investigates acute GVHD severity and intestinal toxicity in patients undergoing allogeneic HSCT. In 38 patients intestinal permeability was assessed before and after chemotherapy (on days -1, +4, +7 and +14 as related to the stem cell infusion). Cumulative acute GVHD (days 0-100) and clinical intestinal toxicity (days 0-14) were evaluated in parallel. Patients with mild, acute GVHD (grades 0-I) had better-preserved intestinal barrier function (P=0.04) and less pronounced cumulative clinical intestinal toxicity (P=0.02) compared with patients with more severe acute GVHD (grades II IV). Gut toxicity predicts acute GVHD severity. Therefore, gut protective strategies may diminish GVHD severity in allogeneic HSCT patients. PMID- 17415647 TI - Proteomic analysis of protein plugs: causative agent of symptoms in patients with choledochal cyst. AB - Symptoms of choledochal cysts are caused by protein plugs. We performed proteomic analysis of protein plugs to elucidate formation mechanism. Protein plugs were obtained from three pediatric patients with choledochal cyst. Proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel bands common to the samples were excised for mass spectrometry. Mass spectra were compared with the NCBI database for protein identification. Gel bands of protein plug samples were predominant at 14 kilodaltons (kDa), followed by 29 kDa. Four other thin bands were common to the plug samples. Four bands (including 14 and 29 kDa) were identified as lithostathine, and one band as serum albumin. Plugs consisted mostly of lithostathine, a protein secreted by pancreatic acinar cells into pancreatic juice. The mechanism involves trypsinogen and lithostathine regurgitating into the cyst through an aberrant union of pancreaticobiliary ducts. Activated trypsin cleaves soluble lithostathine into insoluble forms that aggregate to form plugs. PMID- 17415648 TI - Living donor liver transplantion for primary biliary cirrhosis with autoimmune hemolytic anemia: a case report. PMID- 17415649 TI - Role of platelet-activating factor in acid-induced esophageal mucosal injury. AB - Studies on the pathophysiology of reflux esophagitis have focused on the associated motility and/or structural abnormalities, with relatively little attention directed to inflammatory mediators involved in the acid-induced mucosal injury. Mast cells line the subepithelial lamina propria in both humans and the opossum model, and are ideally positioned to respond to luminal agents that cross the mucosal barrier. To determine whether certain mast cell mediators are involved in acid-induced mucosal injury, epithelial injury scores following 60 min of luminal perfusion of the opossum esophagus with 100 mM HCl were compared in the presence and absence of two different mast cell stabilizers (disodium cromoglycate and doxantrazole) or the selective platelet-activating factor antagonist TCV-309. In control animals acid perfusion caused release of PAF and significant epithelial injury, characterized by epithelial sloughing and cleft formation. This injury was unaffected by pretreatment with disodium cromoglycate or doxantrazole but was completely prevented by TCV-309 (histology damage score, 2.40+/-0.28 in controls vs 0.50 +/- 0.14 in TCV-309-treated animals). These studies suggest that platelet-activating factor is an important mediator of acid induced esophageal mucosal damage. PMID- 17415650 TI - Endoscopic resection of a duodenal gangliocytic paraganglioma. AB - A gangliocytic paraganglioma (GP) is an extremely rare neurogenic tumor nearly located in the second portion of the duodenum and has been regarded as benign. We report a case of duodenal GP in a 53-year-old man. The GP arose in the second portion of the duodenum and was shown by histological examination to consist of epithelioid cells, spindle cells, and ganglion-like cells. An endoscopic ultrasonography showed a round, well-demarcated, inhomogeneous, submucosal tumor in the 3rd-4th layer. We resected it by endoscopic mucosal resection. PMID- 17415651 TI - A rare association of primary biliary cirrhosis with antiphospholipid [corrected] antibody syndrome. PMID- 17415652 TI - Symptomatic and virological response to antiviral therapy in hepatitis C associated with extrahepatic complications of cryoglobulimia. AB - Mixed cryoglobulins are detected in 50% of patients with hepatitis C; fortunately, few have vasculitis affecting skin, peripheral nerves, kidneys, and synovia. This study was designed to identify the natural history of symptomatic cryoglobulinemia and evaluate the response to antiviral therapy. Patients with hepatitis C complicated by symptomatic cryoglobulinemia were assessed for their disease manifestations and response to antiviral therapy. Of 83 patients identified, 56 patients with a minimum of 12 months follow-up were reviewed. Manifestations included dermatologic (75%), rheumatologic (57%), neurologic (34%), and renal (proteinuria 25%). Antiviral therapy was given to 38, of whom 9 were retreated for symptomatic and/or virological nonresponse. Antiviral therapy included interferon monotherapy (n= 8), pegylated-interferon monotherapy (n= 5), consensus-interferon (n= 2), interferon + ribavirin (n= 18), and pegylated interferon + ribavirin (n= 14). Treatment provided sustained symptomatic response in 31 (82%) and virological response in 16 (42%) patients. Symptomatic cryoglobulinemia responds well to antiviral therapy, even when virological response is not achieved. PMID- 17415653 TI - Masters Allen syndrome: a review. PMID- 17415654 TI - Alterations of Toll-like receptor 4 expression on peripheral blood monocytes during the early stage of human acute pancreatitis. AB - We sought to study Toll-like receptor 4 (TLR4) expression on peripheral blood mononuclear cells (PBMCs) during the early stage of human acute pancreatitis (AP). Thirty consecutive patients with acute pancreatitis admitted within 24 hr of onset of abdominal pain were enrolled prospectively in this study. Blood samples were taken by venipuncture at admission and on the third and seventh days after admission. PBMCs were isolated, and TLR4 and CD14 expression on PBMCs was detected by flow cytometer. Serum tumor necrosis factor (TNF)-alpha, interleukin, and lipase were detected simultaneously. Relations among these parameters were analysis. In mild AP, TLR4 expression increased on the first day of admission and then continued to decline for several days. On the seventh day, TLR4 expression was almost normal compared with that of the normal control. The alteration of serum TNF-alpha was coincidence with TLR4. We conclude that mononuclear macrophages might be ignited through TLR4 (the gatekeeper of the innate immune system) and lead to production of TNF-alpha. PMID- 17415655 TI - Amelioration of steatohepatitis with pentoxifylline in a novel nonalcoholic steatohepatitis model induced by high-fat diet. AB - We sought to evaluate the effects of pentoxifylline (PTX) on steatohepatitis in a novel experimental nonalcoholic steatohepatitis (NASH) model induced by a high fat diet (HFD). Thirty-three male Sprague-Dawley rats were randomly divided into 3 groups. The first group received only standard rat diet (control group); groups 2 (placebo group) and 3 were given HFD, ad libitum. After week 4, 0.5 mL of physiologic serum was injected subcutaneously to the placebo group and 50 mg/kg/d PTX was given intraperitoneally to the third group (group PTX). After 6 weeks all rats were humanely killed. Serum biochemistry, tumor necrosis factor-alpha (TNF alpha), plasma, and liver tissue malondialdehyde (MDA) were analyzed. Histopathologically, steatosis, ballooning degeneration, inflammation, and fibrosis were determined. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, plasma and liver tissue MDA, and plasma TNF-alpha levels were significantly higher in placebo group than in the control group. Tumor growth factor-beta levels, however, were comparable in the placebo and control groups. On histopathologic examination, steatosis, inflammatory cells per square millimeter, and ballooning degeneration were significantly higher in the placebo group than in the control group. No fibrosis or Mallory bodies were found in the placebo group. AST, ALT, plasma and liver tissue MDA, and plasma TNF-alpha levels were significantly lower in PTX group compared to the placebo group. Histopathologically, steatosis, mean number of inflammatory cells/mm(2) and ballooning degeneration in PTX group were also significantly lower than in the placebo group. In conclusion, PTX strikingly ameliorates steatohepatitis in this novel NASH model not only by inhibiting the TNF-alpha but also suppressing the oxidative stress markers. PMID- 17415656 TI - Alterations in epithelial and mesenchymal intestinal gene expression during doxorubicin-induced mucositis in mice. AB - In the current study we aimed to gain insight into epithelial-mesenchymal cross talk and progenitor compartment modulation during doxorubicin (DOX)-induced mucositis in mice. Intestinal segments were collected on various days after DOX treatment. DOX-induced damage at day 1-2 was characterized by increased epithelial proliferation and apoptosis and a decrease in the expression of epithelial differentiation markers. Concurrently, T-cell factor-4 (TCF4) levels increased and the epithelial differentiation enhancing factor, bone morphogenic protein-4 (BMP4), decreased. During severe damage (day 3), BMP4 levels were significantly increased, which inversely correlated with epithelial proliferation. At the same time, the expression of the epithelial differentiation markers was increasing again. At day 7, BMP4 levels were down-regulated, while the levels of the epithelial differentiation markers and TCF4 were normalized again. These data suggest that in response to DOX-induced damage, BMP4 and TCF4 are modulated in such a way that homeostasis of the progenitor compartment is partly preserved. PMID- 17415657 TI - Re: Severe toxic hepatitis. PMID- 17415658 TI - Multiple myeloma presenting with malabsorption. PMID- 17415659 TI - Enhanced sensitivity to CD95-induced apoptosis in ob/ob mice. AB - Hepatocyte apoptosis was recently described for NASH patients. The pathomechanisms are incompletely understood, but upregulation of the death receptor Fas was detectable on hepatocytes of NASH patients. We analyzed the sensitivity of fatty liver against CD95/Fas-mediated apoptotic cell death by injection of agonistic anti-Fas antibody (Jo2) in obese ob/ob mice and lean control animals. Ob/ob mice died within 12 hrs, whereas control animals survived. Liver enzymes were significantly increased compared to those in control mice (P < 0.001). Histological analysis and also TUNEL assay of liver sections from ob/ob mice exhibited massive liver injury. Activity of caspase 3 was significantly more enhanced in livers of ob/ob mice after Jo2 challenge. The increased sensitivity was confirmed in vitro by using ob/ob-derived primary hepatocytes. CD95 expression was similar in ob/ob and control mice. However, hepatocytes from ob/ob mice revealed a decreased mitochondrial membrane potential, suggesting that mitochondria play a potential role in this increased susceptibility. PMID- 17415660 TI - In vitro characterization of a compliant biodegradable scaffold with a novel bioreactor system. AB - The influence of scaffold compliance on blood vessel tissue engineering remains unclear and compliance mismatch issues are important to an in vivo tissue engineering approach. We have designed and constructed a modular bioreactor system that is capable of imparting pulsatile fluid flow while simultaneously measuring vessel distension with fluid pressure changes in real time. The setup uses a pneumatic PID control system to generate variable fluid pressure profiles via LabVIEW and an LED micrometer to monitor vessel distension to an accuracy of +/-2 microm. The bioreactor was used to measure the compliance of elastomeric poly(1,8-octanediol citrate) (POC) scaffolds over physiologically relevant pressure ranges. The compliance of POC scaffolds could be adjusted by changing polymerization conditions resulting in scaffolds with compliance values that ranged from 3.8 +/- 0.2 to 15.6 +/- 4.6%/mmHg x 10(-2), depending on the distension pressures applied. Furthermore, scaffolds that were incubated in phosphate-buffered saline for 4 weeks exhibited a linear increase in compliance (2.6 +/- 0.9 to 7.7 +/- 1.2%/mmHg x 10(-2)) and were able to withstand normal physiological blood pressure without bursting. The ability to tailor scaffold compliance and easily measure vessel compliance in real time in vitro will improve our understanding of the role of scaffold compliance on vascular cell processes. PMID- 17415661 TI - A nonlinear model of cardiac autonomic control in obstructive sleep apnea syndrome. AB - Using the Volterra-Wiener approach, we employed a minimal model to quantitatively characterize the linear and nonlinear effects of respiration (RCC) and arterial blood pressure (ABR) on heart rate variability (HRV) in normal controls and subjects with moderate-to-severe obstructive sleep apnea syndrome (OSAS). Respiration, R-R interval (RRI), blood pressure (BP) and other polysomnographic variables were recorded in eight normal controls and nine OSAS subjects in wakefulness, Stage 2 and rapid eye-movement sleep. To increase respiratory and cardiovascular variability, a preprogrammed ventilator delivered randomly timed inspiratory pressures that were superimposed on a baseline continuous positive airway pressure. Except for lower resting RRI in OSAS subjects, summary statistical measures of RRI and BP and their variabilities were similar in controls and OSAS. In contrast, RCC and ABR gains were significantly lower in OSAS. Nonlinear ABR gain and the interaction between respiration and blood pressure in modulating RRI were substantially reduced in OSAS. ABR gain increased during sleep in controls but remained unchanged in OSAS. These findings suggest that normotensive OSAS subjects have impaired daytime parasympathetic and sympathetic function. Nonlinear minimal modeling of HRV provides a useful, insightful, and comprehensive approach for the detection and assessment of abnormal autonomic function in OSAS. PMID- 17415662 TI - Numerical simulation of asymmetrically altered growth as initiation mechanism of scoliosis. AB - The causes of idiopathic scoliosis are still uncertain; buckling is mentioned often, but never proven. The authors hypothesize another option: unilateral postponement of growth of MM Rotatores or of ligamentum flavum and intertransverse ligament. In this paper, both buckling and the two new theories of scoliotic initiation are studied using a new finite element model that simulates the mechanical behavior of the human spine. This model was validated by the stiffness data of Panjabi et al. (J. Biomech. 9:185-192, 1976). After a small correction of the prestrain of some ligaments and the MM Rotatores the model appeared to be valid. The postponement in growth was translated in the numerical model in an asymmetrical stiffness. The spine was loaded axially and the resulting deformation was analyzed for the presence of the coupling of lateral deviation and axial rotation that is characteristic for scoliosis. Only unilateral postponement of growth of ligamentum flavum and intertransverse ligament appeared to initiate scoliosis. Buckling did not initiate scoliosis. PMID- 17415663 TI - Reactive oxygen species induced by proteasome inhibition in neuronal cells mediate mitochondrial dysfunction and a caspase-independent cell death. AB - While increasing evidence shows that proteasome inhibition triggers oxidative damage, mitochondrial dysfunction and death in neuronal cells, the regulatory relationship among these events is unclear. Using mouse neuronal cells we show that the cytotoxicity induced by mild (0.25 microM) and potent (5.0 microM) doses of the proteasome inhibitor, N-Benzyloxycarbonyl-Ile-Glu (O-t-butyl)-Ala leucinal, (PSI) involved a dose-dependent increase in caspase activation, overproduction of reactive oxygen species (ROS) and a mitochondrial dysfunction manifested by the translocation of the proapoptotic protein, Bax, from the cytoplasm to the mitochondria, membrane depolarization and the release of cytochrome c and the apoptosis inducing factor (AIF) from mitochondria to the cytoplasm and nucleus, respectively. Whereas caspase or Bax inhibition failed to prevent mitochondrial membrane depolarization and neuronal cell death, pretreatments with the antioxidant N-acetyl-L-cysteine (NAC) or overexpression of the antiapoptotic protein Bcl-xL abrogated these events in cells exposed to mild levels of PSI. These findings implicated ROS as a mediator of PSI-induced cytotoxicity. However, depletions in glutathione and Bcl-xL with potent proteasome inhibition exacerbated this response whereupon survival required the cooperative protection of NAC with Bcl-xL overexpression. Collectively, ROS induced by proteasome inhibition mediates a mitochondrial dysfunction in neuronal cells that culminates in death through caspase- and Bax-independent mechanisms. PMID- 17415665 TI - Identification of thermostable beta-xylosidase activities produced by Aspergillus brasiliensis and Aspergillus niger. AB - Twenty Aspergillus strains were evaluated for production of extracellular cellulolytic and xylanolytic activities. Aspergillus brasiliensis, A. niger and A. japonicus produced the highest xylanase activities with the A. brasiliensis and A. niger strains producing thermostable beta-xylosidases. The beta-xylosidase activities of the A. brasiliensis and A. niger strains had similar temperature and pH optima at 75 degrees C and pH 5 and retained 62% and 99%, respectively, of these activities over 1 h at 60 degrees C. At 75 degrees C, these values were 38 and 44%, respectively. Whereas A. niger is a well known enzyme producer, this is the first report of xylanase and thermostable beta-xylosidase production from the newly identified, non-ochratoxin-producing species A. brasiliensis. PMID- 17415666 TI - Regioselective and enzymatic production of gamma-resorcylic acid from resorcinol using recombinant Escherichia coli cells expressing a novel decarboxylase gene. AB - A recombinant Escherichia coli, expressing the rdc gene, which encodes a gamma resorcylic acid decarboxylase (Rdc) reversibly catalyzing regioselective carboxylation of resorcinol derived from Rhizobium radiobacter WU-0108, converted 20 mM resorcinol to gamma-resorcylic acid with a 44% (mol/mol) yield at 30 degrees C for 7 h. The recombinant E. coli cells were recyclable at least five times for use as biocatalysts. PMID- 17415667 TI - A phosphate starvation-induced acid phosphatase from Oryza sativa: phosphate regulation and transgenic expression. AB - A phosphate starvation-induced acid phosphatase cDNA was cloned from the rice, Oryza sativa. The cDNA encoding O. sativa acid phosphatase (OsACP1) has 1100 bp with an open reading frame of 274 amino acid residues. The deduced amino acid sequence of OsACP1 cDNA showed 53% identity to tomato acid phosphatase and 46-50% identity to several other plant phosphatases. OsACP1 expression was up-regulated in the rice plant and in cell culture in the absence of phosphate (Pi). The induced expression of OsACP1 was a specific response to Pi starvation, and was not affected by the deprivation of other nutrients. OsACP1 expression was responsive to the level of Pi supply, with transcripts of OsACP1 being abundant in Pi-deprived root. The OsACP1 cDNA was expressed as a 30 kDa polypeptide in baculovirus-infected insect Sf9 cells. In addition, the OsACP1 gene was introduced into Arabidopsis via Agrobacterium-mediated transformation. Functional expression of the OsACP1 gene in the transgenic Arabidopsis lines was confirmed by Northern blot and Western blot analyses, as well as phosphatase activity assays. These results suggest that the OsACP1 gene can be used to develop new transgenic dicotyledonous plants able to adapt to Pi-deficient conditions. PMID- 17415668 TI - Assignment of 1H, 13C, and 15N resonances for SF2 RNA recognition motif 2. PMID- 17415669 TI - 1H, 13C and 15N resonance assignments of rabbit prion protein (91-228). PMID- 17415670 TI - Accumulation of vitamin E in potato (Solanum tuberosum) tubers. AB - Vitamin E (tocopherol) is a powerful antioxidant essential for human health and synthesized only by photosynthetic organisms. The effects of over-expression of tocopherol biosynthetic enzymes have been studied in leaves and seeds, but not in a non-photosynthetic, below-ground plant organ. Genetic and molecular approaches were used to determine if increased levels of tocopherols can be accumulated in potato (Solanum tuberosum L.) tubers through metabolic engineering. Two transgenes were constitutively over-expressed in potato: Arabidopsis thaliana p hydroxyphenylpyruvate dioxygenase (At-HPPD) and A. thaliana homogentisate phytyltransferase (At-HPT). alpha-Tocopherol levels in the transgenic plants were determined by high-performance liquid chromatography. In potato tubers, over expression of At-HPPD resulted in a maximum 266% increase in alpha-tocopherol, and over-expression of At-HPT yielded a 106% increase. However, tubers from transgenic plants still accumulated approximately 10- and 100-fold less alpha tocopherol than leaves or seeds, respectively. The results indicate that physiological and regulatory constraints may be the most limiting factors for tocopherol accumulation in potato tubers. Studying regulation and induction of tocopherol biosynthesis should reveal approaches to more effectively engineer crops with enhanced tocopherol content. PMID- 17415671 TI - Over-expression of an arabidopsis family A sucrose phosphate synthase (SPS) gene alters plant growth and fibre development. AB - The objective of this study was to manipulate the intracellular pools of sucrose by differentially expressing exogenous sucrose phosphate synthase (SPS) and investigating its role in regulating plant growth and fibre development. Tobacco (Nicotiana tabacum cv. Xanthi) plants were transformed with an arabidopsis SPS gene under the regulation of the ubiquitously expressed tandem repeat of the 35S cauliflower mosaic virus promoter, and subject to growth trials and fibre characterization. It was apparent that over-expression of SPS resulted in substantially elevated concentrations of sink sucrose pools compared to wild-type plants, while source tissue sucrose pools remained the same. All transformed plants had significantly increased stem height, which was ascribed to internode elongation, and greater stem diameters, longer fibers and increased total dry biomass relative to the control plants. Difference in the chemical composition of either the storage or structural carbohydrates of the wild-type and SPS transgenic lines were only minor. The correlation between increased stem sucrose content and plant phenotypes with elevated SPS gene expression confirm a role for sucrose availability in controlling plant growth and fibre elongation. PMID- 17415672 TI - Adopting the good reFLEXes when generating conditional alterations in the mouse genome. AB - Major advances have been made in the use of the Cre/loxP system for conditional gene targeting in the mouse. By combining the ability of Cre recombinase to invert or excise a DNA fragment, depending upon the orientation of the flanking loxP sites, and the use of wild-type loxP and variant lox511 sites, we devised an efficient and reliable Cre-mediated genetic switch, called FLEX, through which expression of a given gene can be turned off, while expression of another one can be simultaneously turned on. We discuss how this innovative, flexible and powerful approach, which virtually adapts to any kind of site-specific recombinase (e.g., Cre and Flp recombinases), can be used to easily generate, even at high throughput and genome wide scale, many genetic modifications in a conditional manner, including those which were considered as difficult or impossible to achieve. PMID- 17415673 TI - Bitrophic and tritrophic effects of Bt Cry3A transgenic potato on beneficial, non target, beetles. AB - Insect-resistant transgenic plants have been suggested to have unpredictable effects on the biodiversity of the agro-ecosystem, including potential effects on insect natural enemies, beneficial in control of crop pests. Whilst carnivorous as adults, many of these predators may also consume plant tissues, in particular plant pollen and nectar. Coleoptera are important in terms of agro-ecological research not only because of the large number of species in this order, but also because of their role as biological control agents. Thus any detrimental impact on this group of insects would be highly undesirable. The effects of potato expressing the coleopteran-specific Bacillus thuringiensis delta-endotoxin Cry3A (Bt Cry3A) on the ladybird beetle Harmonia axyridis and the carabid beetle Nebria brevicollis were investigated via the bitrophic interaction of the adult ladybird with potato flowers and the tritrophic interaction of the carabid consuming a non target potato pest. Immunoassays confirmed accumulation of the transgene product in potato leaves and floral tissues (at levels of up to 0.01% (pollen) and 0.0285% (anthers) of total soluble protein). Despite H. axyridis and N. brevicollis belonging to the targeted insect order, no significant effects upon survival or overall body mass change of either beetle were observed. Furthermore, Bt Cry3A had no detrimental effects on reproductive fitness of either beetle species, either in terms of fecundity or subsequent egg viability. Behavioural analysis revealed no significant impact of Bt Cry3A on beetle activity or locomoter behaviour. Ligand blots indicate that this is due to either the absence of Bt-binding sites in brush border membrane vesicles (BBMV) isolated from Nebria brevicollis, or in the case of Harmonia axyridis, the binding did not functionally lead to behavioural or physical effects. PMID- 17415674 TI - A germline transgenic silkworm that secretes recombinant proteins in the sericin layer of cocoon. AB - A silk thread of the silkworm, Bombyx mori, is composed of the insoluble inner fibroin and the hydrophilic outer sericin layer, which are synthesized in the posterior and middle silk gland (MSG), respectively. This study aimed to develop a novel sericin 1 gene (ser1) promoter-driven recombinant expression system using transgenic silkworms, in which recombinant proteins are synthesized in MSG and secreted into the sericin layer. To obtain a high level of gene expression, we tested whether a baculovirus-derived enhancer, hr3, and a trans-regulator, IE1, are capable of stimulating the transcriptional activity of the ser1 promoter, using a transient gene expression system. The results showed that hr3 and IE1 cooperatively increased the ser1 promoter activity more than 30-fold. Then, transgenic silkworms were generated which expressed the EGFP with the signal peptide in MSG under the control of the hr3-linked ser1 promoter and IE1 gene. The silkworms exclusively secreted the EGFP into the sericin layer of cocoons as predicted. The expressed EGFP was extractable from cocoons through a simple procedure with neutral pH buffer solution. The expression system developed in this study enables us to produce recombinant proteins in bulk that can be easily extracted and purified. PMID- 17415675 TI - Stress inducible expression of the DREB1A transcription factor from xeric, Hordeum spontaneum L. in turf and forage grass (Paspalum notatum Flugge) enhances abiotic stress tolerance. AB - The dehydration-responsive element binding proteins (DREB1)/C-repeat (CRT) binding factors (CBF) function as transcription activators and bind to the DRE/CRT cis-acting element commonly present in the promoters of abiotic stress regulated genes. A DREB1A transcription factor ortholog was isolated from a xeric, wild barley (Hordeum spontaneum L.) accession, originating from the Negev desert. Sequence comparison revealed a very high degree of sequence conservation of HsDREB1A to the published barley (Hordeum vulgare L.) DREB1A. Constitutive expression of the HsDREB1A gene was able to trans-activate a reporter gene under transcriptional control of the stress-inducible HVA1s and Dhn8 promoters. HsDREB1A was subcloned under transcriptional control of the stress-inducible barley HVA1s promoter and introduced into the apomictic bahiagrass (Paspalum notatum Flugge) cultivar 'Argentine'. HsDREB1A integration and stress inducible expression was detected in primary transgenic bahiagrass plants and apomictic seed progeny by Southern blot, RT-PCR and northern blot analysis respectively. Transgenic bahiagrass plants with stress-inducible expression of HsDREB1A survived severe salt stress and repeated cycles of severe dehydration stress under controlled environment conditions, in contrast to non-transgenic plants. The observed abiotic stress tolerance is very desirable in turf and forage grasses like bahiagrass, where seasonal droughts and irrigation restrictions affect establishment, persistence or productivity of this perennial crop. PMID- 17415676 TI - PSA--"prostate-specific antigen" or "patient stress & anxiety": a questionnaire study. PMID- 17415677 TI - Cell death in the Purkinje cells of the cerebellum of senescence accelerated mouse (SAMP(8)). AB - The cerebella of SAMP(8) (accelerated aging mouse) and SAMR(1) controls were analyzed by Western Blotting of tyrosine hydroxylase and choline acetyltransferase, as well as by TUNEL and histological silver staining. Both tyrosine hydroxylase and choline acetyltransferase levels were higher in SAMR(1) than in SAMP(8). There was also an age-related decrease in enzyme levels in SAMP(8), with the reduction of tyrosine hydroxylase being more apparent. Concomitantly, there was an age-related increase of apoptosis in the medial neocerebellum and the vermis as revealed by TUNEL, with changes being significant in the SAMP(8) strain. Histologically, some Purkinje cells appeared to disappear during aging. Taken together, the data suggests that the aging SAMP(8) strain displays differential Purkinje cell death in the medial cerebellum and that some of the dying cells are likely to be catecholaminergic. PMID- 17415678 TI - Healthy aging: regulation of the metabolome by cellular redox modulation and prooxidant signaling systems: the essential roles of superoxide anion and hydrogen peroxide. AB - The production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) has long been proposed as leading to random deleterious modification of macromolecules with an associated progressive development of age associated systemic disease. ROS and RNS formation has been posited as a major contributor to the aging process. On the contrary, this review presents evidence that superoxide anion (and hydrogen peroxide) and nitric oxide (and peroxynitrite) constitute regulated prooxidant second messenger systems, with specific sub cellular locales of production and are essential for normal metabolome and physiological function. The role of these second messengers in the regulation of the metabolome is discussed in terms of radical formation as an essential contributor to the physiologically normal regulation of sub-cellular bioenergy systems; proteolysis regulation; transcription activation; enzyme activation; mitochondrial DNA changes; redox regulation of metabolism and cell differentiation; the concept that orally administered small molecule antioxidant therapy is a chimera. The formation of superoxide anion/hydrogen peroxide and nitric oxide do not conditionally lead to random macromolecular damage; under normal physiological conditions their production is actually regulated consistent with their second messenger roles. PMID- 17415679 TI - Nonclassical export pathway: overexpression of NCE102 reduces protein and DNA damage and prolongs lifespan in an SGS1 deficient Saccharomyces cerevisiae. AB - In this study, we used our recently developed screening method, Bud-Scar-based Screening (BSS), to screen a yeast cDNA expression library in an SGS1 deletion BY4742 yeast strain. One gene involved in a nonclassical export pathway, NCE102, was found to extend the life span of Deltasgs1 yeast. Deletion of NCE102 in a wild type yeast strain increased its sensitivity to oxidative stress upon diethylmaleate (DEM) treatment but did not shorten its lifespan, indicating that this gene is not essential in determining yeast lifespan. Transformation of NCE102 into either Deltance102 or Deltasgs1 strains could rescue its tolerance to DEM stress, indicating that NCE102 is protective during oxidative stress. Moreover, overexpression of NCE102 in Deltasgs1 strain leads to reduced protein damage. However, overexpression of NCE102 in wild type yeast strain BY4742 neither protected against oxidative stress due to DEM nor extended yeast lifespan compared to its parental wild type strain, indicating that nonclassical export is redundant and DNA repair is fully sufficient in the wild type strain. We therefore demonstrate that a nonclassical export pathway functions as an alternative clearance/detoxification pathway to eliminate damaged material, when the basic repair pathway is not sufficient. PMID- 17415680 TI - Age-dependent differential expression and activity of rat liver cytosolic inorganic pyrophosphatase gene. AB - Besides epigenetic factors, the genetic make-up and differential gene expression not only determines aging and disease susceptibility but also the functional activity of cells in an individual. Analysis of a variety of mammalian tissues revealed that the age-associated differentially expressed genes mainly belong to inflammation, stress, and metabolism. Intracellular PPi is a by-product of multiple biosynthetic reactions and its hydrolysis by cytosolic inorganic pyrophosphatase (iPPase) has long been considered as an important homeostatic mechanism favoring biosynthesis. In this paper we report an age-associated increase ( approximately 2-fold) in the expression of rat liver cytosolic iPPase gene by differential display PCR and northern blot analysis. Expression profiling of iPPase by RNA slot blot analysis in several other tissues revealed no significant change with aging. A comparative spectrophotometric and in-gel analysis of iPPase activity in whole cell lysate (WCL) of liver, brain, skeletal muscle, heart, spleen and kidney exhibited that liver of old rats (24 months ) has approximately 2-fold more activity than the adult (4 months) ones and also its activity is highest among the tissues. The specificity of iPPase activity in the spectrophotometric assay and in-gel analysis was confirmed by specific iPPase inhibitors like CaCl(2) and NaF. PMID- 17415682 TI - A novel gene STYK1/NOK is upregulated in estrogen receptor-alpha negative estrogen receptor-beta positive breast cancer cells following estrogen treatment. AB - The human STYK1/NOK protein is approximately 30-35% similar to mouse fibroblast growth factor receptor 3 and a kinase homologue in D. melanogaster in the tyrosine protein kinase region. STYK1/NOK was identified as being up regulated in MDA-MB-231, an estrogen receptor-alpha negative breast cancer cell line, following 12 h of estrogen treatment at 1x10(-9) M. On further investigation of STYK1/NOK in estrogen treated cell line MDA-MB-231, STYK1/NOK was up regulated at 6 h post treatment when compared to untreated cells. We also investigated the expression levels of STYK1/NOK in other breast cancer cell lines MCF-7, MDA-MB 231, BT-549, and MDA-MB-435S using QRT-PCR. In addition, the analysis of message accumulation was increased with other synthetic estrogen response modifiers. We propose that the regulation of STYK1/NOK is achieved independent of ERalpha and suggests further investigation to the relevance of this kinase in breast cancer progression. PMID- 17415683 TI - US and UK versions of the EQ-5D preference weights: does choice of preference weights make a difference? AB - BACKGROUND: Most US studies that estimate EQ-5D index score generally apply the UK preference weights. We compared the validity of a newly-developed US weights to the UK weights for use of EQ-5D as a measure of health-related quality of life. METHODS: Data were collected from a randomized clinical trial for patients with HIV (n = 1,126) in the US. Convergent validity was examined by comparing Pearson correlations of EQ-5D index scores with the MOS-HIV Health Survey scale scores and Physical and Mental Health Summary (PHS, MHS) scores using the US and UK weights. Known-groups validity of EQ-5D US versus UK index scores was compared using clinical variables (CD4+ cell count and HIV viral load), and the MOS-HIV PHS and MHS. Score changes in the EQ-5D index from baseline to week 50 were examined using effect size (ES) estimates. RESULTS: The mean EQ-5D index scores was slightly higher using US weights than UK weights (0.87 vs. 0.84, respectively). The correlation coefficient for EQ-5D utilities using the US and UK weights was 0.98. The correlations of EQ-5D index scores with the MOS-HIV scores were moderate and similar using the US and UK weights. The EQ-5D index scores discriminated equally well for both versions between levels of CD4+ count, HIV viral load, and PHS and MHS scores (P < 0.05), suggesting equivalent known groups validity. The changes in EQ-5D index scores from baseline to week 50 were similar for both versions (ES: 0.21 vs. 0.22 for US and UK, respectively), suggesting equivalent responsiveness to score changes. CONCLUSIONS: EQ-5D index scores generated using UK and US preference weights showed equivalent psychometric properties. For assessing treatment benefit in a single population, the use of either the UK or US weights as a measure of HRQOL will not change inferences. However, for comparisons across US and UK populations, the choice between these two weights should be based on their relevance to the study population. PMID- 17415684 TI - Hysteroscopic findings in women at risk of HNPCC. Results of a prospective observational study. AB - OBJECTIVES: To report the feasibility and results of diagnostic hysteroscopy in women at risk of HNPCC. METHODS: Fifty-seven women with mismatch repair gene mutations (n = 11) or Amsterdam II criteria (n = 46) were followed-up prospectively from January 1999 to March 2005. Flexible hysteroscopy was performed once a year. The endometrium was sampled routinely. RESULTS: Of 91 attempted hysteroscopies, 10 failed. The endometrial mucosa appeared normal in 34 cases. Polyps were seen in 12 cases, atrophy in 11, hypertrophy in 10, and fibroids in 7; two hysteroscopies suspected malignancy. A micropolypoid appearance was visualized during five hysteroscopies (5/81, 6%). Of the 86 endometrial biopsy attempts, 64 were successful and showed atrophy (n = 14), proliferation (n = 12), secretion (n = 27), polyps (n = 6), simple hyperplasia without atypia (n = 3), or cancer (n = 2). Micropolypoid appearance was not associated with a specific histological pattern. Operative hysteroscopy was done in 24 cases; in two patients with apparently benign focal lesions the results showed simple hyperplasia without atypias. Five patients underwent hysterectomy (simple hyperplasia without atypias, n = 2; endometrioid adenocarcinoma, n = 2; or secretory mucosa, n = 1). This study led to diagnosis of endometrial simple hyperplasia in 6% of cases and of cancer in 3%. CONCLUSIONS: In patients at risk of HNPCC, hysteroscopy appears feasible to screen endometrial pathology. Two cancers have been diagnosed over 91 patient-years at risk. Hysteroscopy should be compared to sonography as a screening tool in women at risk of HNPCC. PMID- 17415685 TI - Lateral canthal dynamics, correlation with periorbital anthropometric measurements, and effect of age and sleep preference side on eyelid metrics and lateral canthal tendon. AB - PURPOSE: To evaluate dynamic and static properties of lateral canthal tendon and involutional periorbital anthropometric and lateral canthal changes with any possible effect of sleep reference side on these changes. METHODS: Ninety-two healthy adult subjects with a mean age of 43.5 years were enrolled in the study. Lateral canthal tendon lengths (LCT), canthal movement amplitudes, and other periorbital anthropometric parameters were measured. Any effects of age and sleep preference side on anthropometric and particularly lateral canthal tendon measurements were evaluated. Pearson correlation analysis, one-way analysis of variance, and t-test for paired samples were used for statistical evaluation. RESULTS: LCT length was weakly correlated with age, lower lid tractability, horizontal palpebral fissure length, and interpupillary distance and also inversely correlated with margin reflex distance, but not with other measurements. Age was also correlated with lower lid tractability, and with the ratio of LCT to the horizontal palpebral fissure length (LCT-HPFL). No correlation between age and horizontal palpebral fissure length was detected. Age was weakly and inversely correlated with canthal height but not with canthal movement amplitudes. No effect of sleep preference side on LCT length, canthal height, or lower lid tractability was detected. CONCLUSIONS: Lateral canthus has a dynamic structure that is not correlated with any other periorbital anthropometric measurements. LCT length is correlated with lower lid tractability, and increases with age, unlike horizontal palpebral fissure that stays constant with increasing LCT-HPFL ratio. Sleep preference side has no effect on the eyelid metrics and LCT. PMID- 17415686 TI - A comparison of the effect of carbomer-, cellulose- and mineral oil-based artificial tear formulations. AB - PURPOSE: To compare the efficacy, safety, and local tolerance between carbomer based artificial tears, cellulose-, and mineral oil-based artificial tears. METHODS: A randomized, open-label, parallel-group comparative 28-day study was designed for 67 patients who were randomized into three treatment groups. Measurements included the scoring of total subjective symptoms and objective signs, Schirmer-Jones test values, and tear break-up time (BUT) at baseline, and after 2 and 4 weeks of treatment. Safety of study treatment was also assessed. Outcomes measured at baseline and 2 and 4 weeks follow-up included the scoring of total subjective symptoms and objective signs, Schirmer-Jones test values, and tear BUT, subjective assessments, and safety. RESULTS: There were no differences regarding total scores, Schirmer-Jones test, or tear BUT at baseline among these three groups at 2 and 4 weeks. Patients in all three treatment groups experienced a significant improvement from baseline in total scores and Schirmer-Jones test values after treatment. Subjective assessment was better with carbomer-based treatment. CONCLUSIONS: Each artificial tear formulation successfully relieved symptoms and signs of keratoconjunctivitis sicca. The tolerance of carbomer-based artificial tears was comparable to that of cellulose- and mineral oil-based artificial tears. PMID- 17415687 TI - Different conjunctival adaptive response in patients with aqueous-deficient and with mucous-deficient dry eyes. AB - PURPOSE: To describe the different cellular adaptive patterns found in the conjunctival epithelium from patients with aqueous-deficient and mucous-deficient dry eyes. METHODS: The authors studied different conjunctival areas, by impression cytology and by biopsy, 50 eyes with facial nerve paralysis (FNP), 50 eyes with ocular cicatricial pemphigoid (OCP), and 50 eyes from patients with primarily Sjogren syndrome (1SS). RESULTS: Eyes with FNP from the first clinical grade showed a progressive alteration of the nonsecretory cells, with a significant decrease in density goblet cells, generally with a PAS-positive staining. Eyes with OCP, during clinical grades 1 and 2, showed a slow deterioration of the nonsecretory cells; but from clinical grade 3, there was a significant increase of the cellular size and the thickness of the conjunctiva. Goblet cells showed a significant decrease in density from clinical grade 1, generally with a PAS-negative staining. Eyes with 1SS during clinical grades 1 and 2 showed a progressive alteration of the nonsecretory cells, with a significant decrease in density goblet cells, and a PAS-positive staining. From clinical grade 3 appeared a significant increase of nonsecretory cellular size and thickness of conjunctiva, with a significant decrease in goblet cell counts, and a PAS-negative staining. CONCLUSIONS: Patients with FNP (a primarily aqueous deficient alteration) follow completely the squamous metaplasia process. Patients with OCP (a primarily mucous-deficient syndrome) have a hypertrophy and hyperplasia process along the ocular surface. Patients with 1SS (a primarily aqueous-deficient and mucin-deficient alteration) have a squamous metaplasia process, but from clinical grade 3 also appears a hypertrophy and hyperplasia process. PMID- 17415688 TI - Safety and efficacy of two ocular anesthetic methods for phacoemulsification: topical anesthesia and viscoanesthesia (VisThesia). AB - PURPOSE: VisThesia is a new ophthalmic viscosurgical device (OVD) which has 1% lidocaine combined with 1.5% sodium hyaluronate. This is a prospective evaluation of the safety and efficacy of VisThesia used in association with phacoemulsification. METHODS: A total of 114 eyes were divided into two groups. Fifty-nine eyes were treated with tetracaine + oxybuprocaine topical anesthesia and DuoVisc OVD and 55 eyes were treated with VisThesia, for use as both topical anesthetic and OVD. Endothelial cell counts were measured at 30 days postoperatively and compared to preoperative baseline values. Pain and discomfort was subjectively evaluated by patients using a visual analog pain scale (0-10). RESULTS: All surgeries were uneventful with no intraoperative or immediate postoperative complications. Patients receiving topical anesthesia had a mean pain score of 1.1+/-6.8 compared to a mean score of 1.3+/-4.6 for patients receiving VisThesia (p=0.59). Postoperatively, endothelial cell loss at 1 month was greater for patients receiving VisThesia (20.32%+/-43.75) than for those receiving the topical anesthetic (8.8%+/-59.6; p<0.0001). CONCLUSIONS: The results from the visual analog pain scale were comparable between groups, showing that VisThesia provides similar pain relief to topical anesthesia. Specular microscopy performed at 30 days postoperatively showed a significantly greater loss of endothelial cells with the use of VisThesia, suggesting that the 1% lidocaine concentration used in VisThesia may be toxic to corneal endothelial cells. PMID- 17415689 TI - Is pulse synchronized pneumotonometry more reproducible than routine pneumotonometry and more in agreement with Goldmann applanation tonometry? AB - PURPOSE: This study compared the variability of intraocular pressure (IOP) measurements taken with pulse synchronized pneumotonometry (PNT) and with routine PNT (without pulse synchronization) and evaluated the agreement between PNT and Goldmann applanation tonometry (GAT). METHODS: In this prospective study, 148 eyes from 78 patients were enrolled. Patients were randomized into two groups. In the first group (A), the sequence of measurements was pulse synchronized PNT, routine PNT, and GAT. In the second group (B), the sequence of measurements was routine PNT, pulse synchronized PNT, and GAT. The mean of three measurements was averaged for PNT and GAT. All the measurements were performed by the same investigator, who was masked to GAT measurements. The mean IOP measurements and intrapatient standard deviations among the three tonometers were compared using analysis of variance measurement. Bland & Altman plots were used to assess the agreement between PNT and GAT. RESULTS: The variability of IOP measurements taken with pulse synchronized PNT was significantly lower than with routine PNT in both groups (1.2+/-0.7 vs 1.3+/-0.7 mmHg in group A and 1.1+/-0.9 vs 1.3+/-1.0 mmHg in group B [p<0.001], respectively). PNT measurements with and without pulse synchronization were on average +/-2 mmHg higher than GAT measurements in both groups (p<0.001). The 95% limits of agreement between PNT and GAT varied between 3.8 and 8.5 mmHg. CONCLUSIONS: Pulse synchronized PNT gives more reproducible measurements than routine PNT. The agreement between PNT and GAT is poor. PMID- 17415690 TI - A 1-year study to compare the efficacy and safety of once-daily travoprost 0.004%/timolol 0.5% to once-daily latanoprost 0.005%/timolol 0.5% in patients with open-angle glaucoma or ocular hypertension. AB - PURPOSE: The objective of the study was to compare the intraocular pressure (IOP) lowering efficacy and safety of travoprost 0.004%/timolol 0.5% ophthalmic solution (Trav/Tim) to latanoprost 0.005%/timolol 0.5% ophthalmic solution (Lat/Tim), dosed once daily in the morning, in patients with open-angle glaucoma (OAG) or ocular hypertension (OH). METHODS: This was a randomized, double-masked, multicenter, parallel group, active-controlled study conducted at 41 sites. At the eligibility visit the patients were randomized (1:1) to the assigned masked medication if they met inclusion/exclusion criteria, and the mean IOP values in the eligible eyes were > or =24 mmHg at 9 AM and > or =21 mmHg at 11 AM and 4 PM. Patients were excluded if the mean IOP in either eye was >36 mmHg. Patients were instructed to administer the assigned medication each morning at 9 AM. During the treatment phase of the study, IOP was measured at 9 AM at week 2, week 6, month 3, and month 9. At the month 6 and month 12 visits, IOP was measured at 9 AM, 11 AM, and 4 PM. Statistical methods included a repeated measures analysis of variance (ANOVA); to test for noninferiority, a 95% confidence interval for the treatment group difference was constructed based on the ANOVA results for each time point at month 12. RESULTS: Patients (n=408) with OAG or OH were enrolled at 41 sites. One patient withdrew prior to receiving medication so 207 in the Trav/Tim group and 200 in the Lat/Tim group were evaluable for safety. Baseline demographic characteristics as well as IOP values showed no statistical differences between the two groups. Trav/Tim provided lower mean IOP values than Lat/Tim that were statistically significant at the week 2 9 AM (p=0.0081), month 6 9 AM (p=0.0056), and month 6 11 AM (p=0.0128) time points and at 9 AM time point pooled across all visits (p=0.0235) when mean IOP was 0.6 mmHg lower in the Trav/Tim group. Treatment-related adverse events were mild in both groups. Although hyperemia was reported from a higher percentage of patients in Trav/Tim group, differences in average hyperemia scores between the two groups were not considered clinically relevant. CONCLUSIONS: Travoprost 0.004%/timolol 0.5% ophthalmic solution produced mean IOP levels that are statistically noninferior to latanoprost 0.005%/timolol 0.5% ophthalmic solution. Furthermore, at 9:00 AM, 24 hours after dosing, IOP was statistically lower for travoprost 0.004%/timolol 0.5% pooled across all visits. Travoprost 0.004%/timolol 0.5% fixed combination ophthalmic solution is an effective treatment for reducing IOP and it is safe and well-tolerated in patients with OAG or OH. PMID- 17415691 TI - Nd:YAG laser iridotomy in the management of secondary glaucoma associated with Behcet's disease. AB - PURPOSE: To report the outcome of Nd:YAG laser iridotomy in the management of secondary glaucoma associated with Behcet's disease (BD). METHODS: In this prospective study, Nd:YAG laser iridotomy was performed on eyes with secondary angle-closure and pupillary block glaucoma associated with BD. The pretreatment and post-treatment intraocular pressures (IOP) and the number of antiglaucoma medications were compared by Mann-Whitney U test. RESULTS: The study consisted of 16 eyes of 11 patients (2 female, 9 male, mean age 39.2+/-8.9 years). Post treatment follow-up ranged from 6 to 36 months (mean 13.8+/-8.9). The mean IOP was 21.6+/-2.5 mmHg on 2.5+/-0.6 medications before iridotomy. IOP reduced to 17.7+/-2.5 mmHg on 1+/-0.6 medications at the first month and 17.1+/-3.2 mmHg on 1.7+/-0.9 medications at the sixth month of treatment. The differences between IOP and number of antiglaucoma medications at baseline and at the sixth month of the treatment was statistically significant (p<0.00001). For four eyes trabeculectomy with mitomycin C and for one eye Ahmed valve implantation were performed in the follow-up period. CONCLUSIONS: Nd:YAG laser iridotomy can provide reduction of IOP and the number of antiglaucoma medications in selected cases with secondary glaucoma associated with Behcet's disease. PMID- 17415692 TI - Comparison of Swedish interactive threshold algorithm and full threshold algorithm for glaucomatous visual field loss. AB - PURPOSE: To compare the prevalence of visual field loss, the sensitivity distribution, and the size and depth of glaucomatous visual field defects using the standard full threshold (FT) and the Swedish interactive threshold algorithm (SITA) standard (SS) procedures in patients with early or suspected glaucoma. METHODS: Automated perimetry findings were retrospectively evaluated in 53 patients (105 eyes) with early or suspected glaucoma. RESULTS: The number of eyes judged to have glaucomatous visual field loss by SS (48 eyes) was significantly larger than what was found with FT (35 eyes), and 70 eyes were classified as pre perimetric glaucoma. In these 70 eyes, there were many locations where the sensitivity was significantly higher with SS than with FT (intrasubject difference), and SS had less intersubject variability than FT at most locations. The cumulative decibel scores at the region of glaucomatous defects were larger with SS (206.2+/-103.3 dB) than with FT (162.1+/-87.5 dB) (p=0.02), which indicated that the depth of defects measured by SS was shallower than that by FT. The sizes of defects were significantly larger with SS (11.2+/-5.6) than with FT (9.7+/-5.1) (p<0.05). CONCLUSIONS: Glaucomatous defects were measured as being significantly shallower and larger with SS than with FT. In addition, the prevalence of visual field defect was higher with SS according to some of the criteria for glaucomatous visual field defects. These results might be related to the fact that SS strategy has a lower variability and to the Bayesian statistical properties of the SITA algorithm. PMID- 17415693 TI - Test-retest variability in visual field testing using frequency doubling technology. AB - PURPOSE: To quantify the magnitude of test-retest variability (TRV) for normal subjects in serial visual fields (VF) using the frequency doubling technology (FDT) instrument. METHODS: Twenty-one healthy adults, aged 23 to 60 years, underwent four serial FDT VF tests, using the full-threshold C-20 program of the Zeiss-Humphrey FDT analyzer, on one randomly chosen eye. The VF tests were spaced 2 to 4 days apart. All subjects performed two preliminary FDT tests in order to minimize any learning effect. Test-retest variability was calculated as the standard deviation of each location's sensitivity value across the four VF tests. RESULTS: Mean TRV (+/-SD) for the entire field was 2.44+/-1.32 dB. Mean TRV (+/ SD) for the superior, inferior, nasal, and temporal hemifields were 2.48+/-1.3, 2.40+/-1.4, 2.40+/-1.3, and 2.48+/-1.3 dB, respectively. Mean TRV (+/-SD) for the foveal location, the 4 central, and the 12 peripheral locations were 2.49+/-1.4, 2.16+/-1.2, and 2.54+/-1.4 dB, respectively. CONCLUSIONS: TRV was found to be rather uniform across the visual field of the commercially available FDT device, with only a mild, clinically insignificant, effect of both eccentricity and age on TRV. Variability in the FDT VF, for normal subjects, was found to be more uniform than that of both standard and short wavelength automated perimetry. In addition, a strong inverse correlation was found, in normal subjects, between the mean sensitivity and TRV. PMID- 17415694 TI - Combined phacoemulsification and nonpenetrating deep sclerectomy in the treatment of chronic angle-closure glaucoma with cataract. AB - PURPOSE: To review the result of nonpenetrating deep sclerectomy (NPDS) combined with phacoemulsification in the treatment of chronic angle-closure glaucoma (CACG) with coexisting cataract. METHODS: This is a retrospective review of 29 eyes of 26 patients who had undergone combined NPDS and phacoemulsification for cataract and CACG between January 2001 and June 2003. The visual acuity, intraocular pressure (IOP), and complications were analyzed. RESULTS: The mean follow-up period was 33.8 months (range 23.3 to 54.0 months). Postoperative visual acuity improved in 21 eyes (72%) and remained the same in 6 eyes (21%). The IOP was reduced significantly from 20.3+/-3.9 mmHg (mean +/- SD) preoperatively to 15.9+/-3.1 mmHg postoperatively at last follow-up visit (p<0.001). The number of antiglaucoma medications was also reduced significantly from 2.9+/-0.8 (mean +/- SD) preoperatively to 1.0+/-1.2 at last follow-up (p<0.001). Fifteen eyes (52%) achieved complete success with IOP < or = 21 mmHg without antiglaucoma medications and 25 eyes (86%) achieved qualified success with IOP < or = 21 mmHg with or without medications at the last follow-up visit. Of the 25 eyes achieving qualified success, 24 (96%) had a reduction in the number of medications. There were 4 failures, defined as uncontrolled IOP requiring further filtering operation or oral drug treatment. Intraoperative complications included one accidental anterior chamber puncture and one iris plug intraoperatively. Postoperative complications included one choroidal effusion, three wound leaks requiring repair, and two punctate epithelial erosions. There was no shallowing of the anterior chamber, hyphema, hypotony, or infection encountered. CONCLUSIONS: Combined NPDS and phacoemulsification could be a safe and effective surgical option for the management of CACG with cataract. PMID- 17415695 TI - Diagnostic accuracy of the Moorfields Regression Analysis using the Heidelberg Retina Tomograph in glaucoma patients with visual field defects. AB - PURPOSE: This study was designed to determine the specificity and sensitivity of the Heidelberg Retina Tomograph I (HRT) using the Moorfields Regression Analysis (MRA) in differentiating healthy from glaucomatous eyes. METHODS: In this cross sectional study, 74 eyes of 37 healthy subjects and 87 eyes of 47 glaucoma patients were examined with Octopus standard automated perimetry and HRT. Only one eye per patient was used for statistical analysis. According to visual field index mean defect (MD) glaucoma patients were divided into three groups with early (MD < 6 dB), moderate (6 dB < MD < 12 dB), and advanced visual field loss (MD > 12 dB). The sensitivity and specificity of optic nerve head examinations using the MRA of HRT were evaluated by two criteria (criteria 1, as diseased if classified by MRA as outside normal limits; criteria 2, as diseased if classified by MRA as borderline). The correlations between the topographic parameters and visual field index MD were measured by correlation coefficient and presented by scatter plot. RESULTS: The specificity and sensitivity of HRT-MRA examination were, respectively, 100% and 68.1% when borderline cases were considered normal (criteria 1) and 97.3% and 85.1% when borderline cases were considered glaucomatous (criteria 2). The sensitivity of the MRA in eyes with different stages of visual field loss was 59.1% for early, 54.5% for moderate, and 92.8% for advanced visual field loss with criteria 1; the figures were, respectively, 81.8%, 72.7%, and 100% with criteria 2. The statistically significant correlations with moderate strength of association (r = 0.40-0.59) were found for rim area, rim volume, cup to disc area ratio, mean retinal nerve fiber layer (RNFL) thickness, and RNFL cross-sectional area. CONCLUSIONS: The MRA showed an excellent specificity and good sensitivity using criteria 2 including as glaucomatous optic discs those classified by MRA as borderline. Although correlations of moderate strength were found between rim area, rim volume, mean RNFL thickness, RNFL cross-sectional area, and visual field index MD, great interindividual variation limits the prediction of one parameter from the other. Therefore, in clinical practice both structural and functional examinations should be performed in order to characterize glaucomatous damage. PMID- 17415696 TI - Sector-based analysis of frequency doubling technology sensitivity and optic nerve head shape parameters. AB - PURPOSE: To evaluate which Heidelberg Retina Tomograph (HRT) parameter is the best predictor of frequency doubling technology (FDT) sensitivity by using a sector based analysis between FDT and optic nerve head shape parameters such as cup shape measure (CSM) and rim area (RA), which have been shown to have the best correlation with FDT indices among all the HRT parameters. METHODS: One eye was randomly chosen from 100 patients with primary open angle glaucoma (abnormal visual field and/or abnormal optic nerve and untreated intraocular pressure above 21 mm Hg). All the patients were examined with Humphrey field analyzer (HFA), program 24-2, SITA standard, FDT program C 20, and HRT. RA and CSM for the HRT analysis and mean deviation, pattern standard deviation, and the sensitivity of each tested point for the FDT test were considered in this study. All the parameters were calculated as both global and sector measurements. Findings were analyzed using Pearson's correlation coefficient and linear regression model. RESULTS: Significant (p<0.001) correlation was found between FDT indices and HRT RA and CSM. Significant (p<0.001) sector correlation was found between FDT sensitivity and RA and CSM, but when a linear regression model was applied, RA was the most predictive parameter of FDT. Temporal CSM was more strongly correlated to FDT fovea sensitivity than temporal RA. Furthermore, sector HRT parameters were better correlated to HFA than to FDT. CONCLUSIONS: In this glaucomatous group, cup shape measure and RA were significantly correlated to FDT indices. RA was more predictive of FDT abnormality than CSM in all the considered sectors except in the temporal one. However, HFA was a stronger predictor of HRT parameters than FDT. PMID- 17415697 TI - Intravitreal bevacizumab therapy for choroidal neovascularization secondary to age-related macular degeneration: 6-month results of an open-label uncontrolled clinical study. AB - PURPOSE: To investigate the 6-month safety and clinical outcomes of intravitreal injections of bevacizumab administered to treat choroidal neovascularization secondary to age-related macular degeneration. METHODS: Twenty-seven patients underwent 1.25 mg intravitreal injections of bevacizumab at baseline. A similar intravitreal injection was administered to all eyes at 1 and 2 month follow-up visits. At baseline and at each follow-up visit (1, 2, 3, and 6 months), patients underwent best-corrected visual acuity (BCVA) measurement, fluorescein angiography, indocyanine green angiography, and optical coherence tomography. Laboratory testing, visual field analyses, and endothelial cell counts were performed at baseline and third and sixth months. RESULTS: At 3 months, the mean BCVA remained substantially stable at 20/100. Mean central retinal thickness (CRT) decreased from 373 to 279 microm (p<0.01). Mean lesion greatest linear dimension (GLD) decreased from 4087 to 3782 microns (p<0.01). At 6 months, mean BCVA slightly decreased from 20/100(-1) to 20/125(-3) (not significant, p=0.40). Mean CRT was still inferior to baseline (305 microm, p<0.01). Mean lesion GLD was 4186 microm, not different from baseline values (p=0.59), but superior to 3-month mean GLD (p<0.01). Significant visual field defects or endothelial cell losses were not detected at 3 and 6 months. Laboratory testing did not reveal any clinically significant deviations compared to baseline values. CONCLUSIONS: Intravitreal therapy using bevacizumab over 6 months showed stabilization of visual acuity and choroidal neovascularization activity; the safety data were convincing. PMID- 17415698 TI - Retinal imaging by spectral optical coherence tomography. AB - PURPOSE: To demonstrate applicability of high speed spectral optical coherence tomography (SOCT) method for imaging retinal pathologies in clinical conditions. METHODS: SOCT was performed in 67 eyes with different macular diseases. Examinations were carried out with the prototype SOCT instrument constructed in the Institute of Physics, Nicolaus Copernicus University, Torun, Poland. A broadband superluminescent diode was used as a light source. RESULTS: The disturbances of retinal layer structure concerning mainly outer segments of photoreceptors were observed in case of central serous chorioretinopathy and choroidal neovascularization in age-related macular degeneration. Large drusen were often related to significant changes of outer nuclear layer thickness and reflectivity. CONCLUSIONS: SOCT detects small disturbances of the retinal structure and helps to precisely determine layers involved in different pathologies. PMID- 17415699 TI - A new method of treating macular holes. AB - PURPOSE: To evaluate the effect of pars plana vitrectomy (PPV) combined with joining of the hole edges on the closure of full-thickness macular holes. METHODS: In a prospective consecutive clinical trial, standard PPV with internal limiting membrane (ILM) maculorhexis was performed in 25 eyes from 25 patients with stage 3 or 4 idiopathic macular holes (Group A). The retina was massaged gently around the hole from the periphery to the center in order to approximate the edges as closely as possible. The edges of the macular hole were then joined with forceps and gently pressed together. At the end of surgery, air was used for intravitreal tamponade. Patients were required to remain in a face-down position for 1 day postoperatively. For the comparison, a retrospective analysis of outcomes of surgical treatment of 27 eyes of 27 patients with stage 3 to 4 idiopathic macular hole (Group B), whose surgery included standard three-port PPV, followed with ILM peeling, was performed. The closed macular holes were categorized into two patterns based on optical coherence tomography: flat/closed and flat/open. RESULTS: The overall closure rate was 92+/-5.4% over a minimum follow-up period of 6 months in Group A, and 86+/-6.2% in Group B. Best-corrected visual acuity improved from 0.1+/-0.014 (ranged from 0.02 to 0.5) before surgery to 0.29+/-0.03 (ranged from 0.2 to 0.7) after surgery in Group A, and from 0.1+/ 0.05 (ranged from 0.05 to 0.4) before surgery to 0.22+/-0.04 (ranged from 0.05 to 0.4) after surgery in Group B. No significant difference was found in absolute light sensitivity of macula, intraocular pressure, or lens opacification. A common postoperative complication in Group A was retinal pigment epitheliopathy, which developed in 18 cases (72%). CONCLUSIONS: Mechanical joining and compression of the retinal edges during surgery for stage 3 or 4 idiopathic macular holes appears to yield a promising anatomic and functional result. PMID- 17415700 TI - Dominant optic atrophy in a Japanese family with OPA1 frameshift mutation (V942fsX966). AB - PURPOSE: The authors report the ophthalmic characteristics of a male proband in a Japanese family with autosomal dominant optic atrophy (DOA) harboring a frameshift mutation in the OPA1 gene. METHODS: Conventional ophthalmologic examinations including static automated perimetry were performed, as well as assessment of the three-generation family history. The peripapillary retinal nerve fiber layer (RNFL) was evaluated using scanning laser polarimetry. Mutation screening of the OPA1 gene was performed with polymerase chain reaction amplification and direct sequencing. RESULTS: A frameshift mutation (p.V942fsX966) was identified in the proband and his mother. In comparison with the adolescent onset of visual loss in the proband and his maternal grandfather, the mother presented with only subtle temporal disc pallor and has never been aware of any visual disturbances. Symmetric thinned peripapillary RNFL was detected in the proband, whose visual field abnormalities were limited to central scotomas and were without mean deviation worsening between 11 to 17 years of age in both eyes. The proband's logMAR visual acuity (0.52 to 0.7) has remained almost unchanged for more than 10 years since initial evaluation at age 10. CONCLUSIONS: The OPA1 mutation may be correlated with slow progression of DOA, and with phenotypic variations within the family. Further study is necessary to determine whether symmetric thinned peripapillary RNFL represents a feature of DOA. PMID- 17415701 TI - Ultrabiomicroscopy (UBM) in flap dislocation following LASIK: a case report. AB - PURPOSE: To describe postoperative laser-assisted in situ keratomileusis (LASIK) flap dislocation occurred after trauma METHODS: Ultrabiomicroscopy (UBM) is used to obtain a high-resolution imaging of the cornea. RESULTS: The UBM results are presented and compared with histologic and confoscan findings CONCLUSIONS: The technique is useful and easy to perform, offering more opportunities to study the anatomical changes in LASK flap dislocation occurred after trauma. PMID- 17415702 TI - Development of early choroidal neovascular membrane in a young myope after LASIK. AB - PURPOSE: To describe development of early choroidal neovascular membrane (CNVM) after laser-assisted in situ keratomileusis (LASIK) procedure in a young myopic patient that was successfully managed by photodynamic therapy (PDT). METHODS: A retrospective interventional case report. RESULTS: A 20-year-old woman with myopic astigmatism underwent uneventful LASIK surgery resulting in best-corrected visual acuity (BCVA) of 20/20 bilaterally. One week later, the patient presented with decreased VA in the right eye and was found to have clinical evidence of central serous chorioretinopathy (CSC). She was treated with systemic corticosteroids without any improvement. Two weeks after LASIK, the patient's VA was 20/200 and clinical evidence of early CNVM was confirmed by fluorescein angiography, indocyanine green angiography, and optical coherence tomography (OCT). PDT resulted in the regression of her CNVM with improvement in her VA and OCT findings. CONCLUSIONS: Systemic corticosteroids may enhance CNVM in patients with LASIK-induced early CNVM. PDT may be effective in the resolution of LASIK induced CNVM. PMID- 17415703 TI - Removal of Thelazia callipaeda from the subconjunctival space. AB - PURPOSE: To report the finding of Thelazia callipaeda within the human subconjunctival space. METHODS: An 81-year-old man with a history of traumatic conjunctival laceration that occurred 2 years previously had white worms in the subconjunctival space of his right eye. RESULTS: Five worms were removed from the subconjunctival space via a local peritomy, since there was no conjunctival laceration noted during the examination. These worms were identified as T callipaeda. CONCLUSIONS: T callipaeda cannot dig holes in the ocular wall due to the lack of hooks or sharp spines within the mouth. Therefore, the authors speculate that these worms entered the subconjunctival space through a conjunctival laceration that had occurred 2 years previously. PMID- 17415704 TI - Intravitreal bevacizumab for neovascular glaucoma following central retinal artery occlusion. AB - PURPOSE: To report a case of neovascular glaucoma due to central retinal artery occlusion treated with a single intravitreal injection of bevacizumab. METHODS: A 68-year-old patient with a 10-week history of central retinal artery occlusion presented with neovascularization of the iris and the angle and intraocular pressure of 30 mm Hg. The patient received a single injection of 1.25 mg bevacizumab in 0.1 mL intravitreally. RESULTS: Iris and angle neovascularization regressed within 48 hours of the injection. Intraocular pressure dropped from 30 to 15 mm Hg, and there was marked improvement in patient comfort. Panretinal photocoagulation was applied 4 weeks after the injection. CONCLUSIONS: Bevacizumab seems to be a useful adjunct to panretinal photocoagulation in the treatment of neovascular glaucoma. PMID- 17415705 TI - Optical coherence tomography study of vitreoretinal interface in full thickness macular hole associated with optic disk pit maculopathy. AB - PURPOSE: To evaluate by optical coherence tomography (OCT) the vitreous involvement in full thickness macular hole associated with optic disk pit maculopathy. METHODS: Two patients with optic disk pit maculopathy and full thickness macular hole underwent OCT for evaluation of vitreous involvement in the pathogenesis of the disease. Fluorescein angiography and fundus photographs were also performed. RESULTS: OCT documented the presence of vitreous traction at the edge of the full thickness macular hole in the first case and on the elevated macula in the second case. Partial posterior vitreous detachment was also noticed. CONCLUSIONS: Previously unreported OCT findings such as vitreous traction at the edge of the macular hole and on the elevated macula were found. OCT provided evidence for vitreous participation in full thickness macular hole associated with optic disk pit maculopathy. OCT was also able to show that the full thickness macular hole and optic disk pit maculopathy were probably caused by similar vitreous tractional forces. PMID- 17415706 TI - Treatment of the mild cognitive impairment (MCI). AB - According to Evidence-Based-Medicine, any proposal for the rationale treatment of mild cognitive impairment (MCI) must be based on the results of double-blind, randomized clinical trials (RCTs). However, since MCI at the present time does not constitute a homogeneous clinical syndrome, it is still inappropriate to propose a specific drug treatment. Moreover, RCTs assessing the therapeutic value of acetylcholinesterase-inhibitors (AChEIs) are negative either trying to improve symptoms (memory performance) or preventing the conversion from MCI to real Alzheimer's Disease (AD). The same negative results were obtained with drugs targeting some systems considered as the early steps of the pathophysiological cascade leading to dementia: non-steroidal anti-inflammatory compounds (rofecoxib), sex steroid hormones (testosterone, estrogens), or antioxidants (tocopherol). Either MCI is considered as the very early phase of development of AD (and then the treatments will aim at preventively antagonizing the hallmarks of the disease) or MCI is a new entity (and then the drugs will target the associated neurochemical disturbances such as tau protein or soluble Abeta oligomers); MCI could also be considered as a monosymptomatic syndrome (amnesia) leading to the development of pure pro-mnestic drugs. These three hypotheses will be presented on the basis of the neurobiology and the pharmacology, and examples of potentially active candidates will be discussed. PMID- 17415708 TI - Identification of a novel noncoding RNA gene, NAMA, that is downregulated in papillary thyroid carcinoma with BRAF mutation and associated with growth arrest. AB - In search of tumor suppressor genes in papillary thyroid carcinoma (PTC), we previously used gene expression profiling to identify genes underexpressed in tumor compared with paired unaffected tissue. While searching for loss of heterozygosity (LOH) in genomic regions harboring candidate tumor suppressor genes, we detected LOH in a approximately 20 kb region around marker D9S176. Several ESTs flanking D9S176 were underexpressed in PTC tumors, and for one of the ESTs, downregulation was highly associated with the activating BRAF mutation V600E, the most common genetic lesion in PTC. A novel gene, NAMA, (noncoding RNA associated with MAP kinase pathway and growth arrest) containing the affected EST was cloned and characterized. NAMA is weakly expressed in several human tissues, and the spliced forms are primarily detected in testis. Several characteristics of NAMA suggest that it is a nonprotein coding but functional RNA; it has no long open reading frames (ORFs); the exons exhibit low sequence identity in the evolutionarily conserved regions; it is inducible by knockdown of BRAF, inhibition of the MAP kinase pathway, growth arrest and DNA damage in cancer cell lines. We suggest that NAMA is a noncoding RNA associated with growth arrest. PMID- 17415709 TI - Expression of B7-H1 in breast cancer patients is strongly associated with high proliferative Ki-67-expressing tumor cells. AB - B7-H1, a co-inhibitory molecule, plays a role in immune escape of tumors. We have shown previously the expression of this molecule in breast cancer patients and demonstrated its association with high histological grade, progesterone and estrogen receptor negative status, all of which are known to have direct impact on cell proliferation. In the present work, we investigated the effect of proliferation, as measured by Ki-67 and mitotic count, on the induction of B7-H1. We used H&E stained sections to score for mitotic count in 69 breast cancer patients. Immunohistochemistry was used to investigate B7-H1 and Ki-67 expression. The relationship between B7-H1 induction and cell proliferation was further investigated in primary cultured cells. B7-H1 expression was recorded in patients with a high mitotic index (p = 0.007). There was a high significant correlation between B7-H1 expression and the presence of the proliferative marker Ki-67 (p < 0.001) indicating the association of proliferation with B7-H1 induction. Furthermore, B7-H1 was gradually induced in proliferating cells of 8/8 primary cell lines as measured by Ki-67 expression. Finally, B7-H1 was downregulated in quiescent cells and upregulated in cells stimulated with a mitogen confirming the association of proliferation with the induction of B7-H1. We have shown for the first time a direct association between proliferation and the expression of B7-H1 in breast cancer patients. The relationship between B7-H1 induction and cell proliferation was also thoroughly investigated in vitro, in which a strong link between B7-H1 expression and the presence of the proliferative Ki-67 marker was clearly demonstrated. PMID- 17415710 TI - Epigenetic silencing of the candidate tumor suppressor gene PROX1 in sporadic breast cancer. AB - Extensive hypermethylation and consecutive transcriptional silencing of tumorsuppressor genes have been documented in multiple tumor entities including breast cancer. In a microarray based genome-wide methylation analysis of five sporadic breast carcinomas we identified a hypermethylated CpG island within the first intron of the prospero related homeobox gene 1 (PROX1). We, therefore, investigated CpG island methylation of PROX1 in a series of 33 pairs of primary breast cancer and corresponding normal tissue samples by bisulfite sequencing and COBRA analyses. Seventeen of these (52%) breast cancer samples revealed a significant accumulation of methylated CpG sites along with a significant reduction of PROX1 transcription compared to normal breast tissues of the same patients. Frequent methylation was also observed in brain metastases from primary breast cancer (21/37 = 57% of cases). Secondary, we analysed 38 brain metastases of primary breast carcinomas and detected a significantly reduced expression of PROX1 compared to normal breast tissue (p < 0.001) and primary breast carcinomas (p < 0.05), respectively. Additionally, treatment of breast cancer cell lines with demethylating agents could reactivate PROX1 transcription. In summary, we have identified PROX1 as a novel target gene that is hypermethylated and transcriptionally silenced in primary and metastatic breast cancer. PMID- 17415711 TI - P53-specific T cell responses in patients with malignant and benign ovarian tumors: implications for p53 based immunotherapy. AB - Despite intensive treatment, 70% of the ovarian cancer patients will develop recurrent disease, emphasizing the need for new approaches such as immunotherapy. A promising antigenic target for immunotherapy in ovarian cancer is the frequently overexpressed p53 protein. The aim of the study was to evaluate the nature and magnitude of the baseline anti-p53 immune response in ovarian cancer patients. P53-specific T cell responses were detected in both half of the ovarian cancer patients as in the group of control subjects, consisting of women with benign ovarian tumors and healthy controls. Importantly, while in the control group p53-specific immunity was detected among the CD45RA(+) naive subset of T cells only, the p53-specific T-cell responses in ovarian cancer patients were also present in the CD45RO(+) memory T-cell subset, suggesting that in the cancer patients sufficient amounts of cancer-derived p53 was presented to induce the formation of a p53-specific memory T-cell response. Further characterization of the p53-specific memory T-cell responses revealed that in addition to the type 1 cytokine IFN-gamma also the type 2 cytokines IL-4 and IL-5, as well as the immunosuppressive cytokine IL-10 were produced. Notably, p53-specific T cells were not only detected in the peripheral blood, but also among tumor infiltrating lymphocytes and in tumor-draining lymph nodes. In conclusion, the existence of a weak mixed T-helper type 1 and 2 p53-specific T-cell repertoire supports the rationale of using p53 long peptides in vaccination strategies aiming at the induction of p53-specific Th1/CTL immunity. PMID- 17415712 TI - Ras-MEK-ERK signaling cascade regulates androgen receptor element-inducible gene transcription and DNA synthesis in prostate cancer cells. AB - Treatment of prostate cancer (CaP) patients frequently involves androgen ablation, but resistance often develops and androgen-insensitive tumors emerge. The molecular basis for the development of refractory CaP that grows in an androgen-independent manner is poorly understood, but alterations in growth factor signaling pathways are likely to be involved. We examined the growth factor modulation of androgen-receptor element (ARE)-inducible luciferase reporter gene activity and consequent DNA synthesis as a measure of proliferative growth in androgen-dependent LNCaP or androgen-independent PC3 or DU145 CaP cells. The synthetic androgen R1881 stimulated ARE-inducible reporter gene activity and prostate-specific antigen expression in LNCaP cells and the MEK/ERK inhibitor U0126 or the anti-androgen bicalutamide (casodex) prevented both of these responses. Activated V12-Ha-Ras expression in LNCaP cells also stimulated ARE-inducible gene transcription, and U0126 or the farnesyltransferase inhibitor FTI-277 but not bicalutamide blocked this. ARE-inducible reporter gene activity was elevated already in PC3 cells, and ERK was constitutively activated in serum starved LNCaP or DU145 cells. U0126 inhibited each of these responses and also inhibited DNA synthesis in all 3 CaP cell lines. These results demonstrate that chronic stimulation of the Ras-MEK-ERK signaling pathway can sustain ARE inducible gene transcription and growth of CaP cells, and suggests that components of this pathway may offer targets for cancer therapy. PMID- 17415713 TI - Expression of oncofetal RNA-binding protein CRD-BP/IMP1 predicts clinical outcome in colon cancer. AB - The oncofetal CRD-BP/IMP1 RNA binding protein regulates posttranscriptionally a handful of RNA transcripts, implicated in cell adhesion and invadopodia formation and was recently identified as a target of the beta-catenin/Tcf transcription factor that is constitutively activated in colorectal carcinomas (CRCs). The expression of CRD-BP/IMP1 was studied in normal adult intestines and CRCs. In normal mucosa, CRD-BP/IMP1 immunoreactivity was observed in few scattered cells located predominantly at or near the bottom of the crypts, whereas in CRCs the protein was detectable in tumor cells of 50% of the specimens analyzed. CRD BP/IMP1 mRNA expression was measured by qRT-PCR in 78 CRCs. Thirty-two (41%) of the specimens were negative or had negligible expression, whereas the remaining forty-six (59%) expressed a wide range of CRD-BP/IMP1 mRNA levels. CRD-BP/IMP1 mRNA expression correlated with that of the putative stem/progenitor cell marker Musashi-1 mRNA (p = 0. 035). CRD-BP/IMP1 positive tumors metastasized and/or recurred more frequently (p = 0.001) and its expression defined a group of patients with shorter survival (p = 0.014). Furthermore, in a multivariate analysis CRD-BP/IMP1 expression was found to be an independent predictor of survival (p = 0.015). For stage I & II patients, the differences in metastasis/recurrence and survival rates remained significant (p = 0.001 and 0.033, respectively). These findings indicate that CRD-BP/IMP1 positive tumors exhibit early disease dissemination and unfavorable prognosis. PMID- 17415714 TI - Socioeconomic inequalities in alcohol related cancer mortality among men: to what extent do they differ between Western European populations? AB - We aim to study socioeconomic inequalities in alcohol related cancers mortality [upper aerodigestive tract (UADT) (oral cavity, pharynx, larynx, oesophagus and liver)] in men and to investigate whether the contribution of these cancers to socioeconomic inequalities in cancer mortality differs within Western Europe. We used longitudinal mortality datasets, including causes of death. Data were collected during the 1990s among men aged 30-74 years in 13 European populations [Madrid, the Basque region, Barcelona, Turin, Switzerland (German and Latin part), France, Belgium (Walloon and Flemish part, Brussels), Norway, Sweden, Finland]. Socioeconomic status was measured using the educational level declared at the census at the beginning of the follow-up period. We conducted Poisson regression analyses and used both relative [Relative index of inequality (RII)] and absolute (mortality rates difference) measures of inequality. For UADT cancers, the RII's were above 3.5 in France, Switzerland (both parts) and Turin whereas for liver cancer they were the highest (around 2.5) in Madrid, France and Turin. The contribution of alcohol related cancer to socioeconomic inequalities in cancer mortality was 29-36% in France and the Spanish populations, 17-23% in Switzerland and Turin, and 5-15% in Belgium and the Nordic countries. We did not observe any correlation between mortality rates differences for lung and UADT cancers, confirming that the pattern found for UADT cancers is not only due to smoking. This study suggests that alcohol use substantially influences socioeconomic inequalities in male cancer mortality in France, Spain and Switzerland but not in the Nordic countries and nor in Belgium. PMID- 17415716 TI - Sunscreen use and increased duration of intentional sun exposure: still a burning issue. AB - Sunscreen use is often proposed for sun protection because of their ability to block UV-induced sunburns (the sun protection factor--SPF). Among suntan seekers, however, risk of cutaneous melanoma may be increased because of extended sun exposure duration. We made a systematic review of the evidence linking sunscreen use to sun exposure duration. Five observational studies found that when sun exposure was associated with willingness to get a tan or to stay longer in the sun (i.e., intentional sun exposure), sunscreen use was associated with duration of sun exposure 13-39% longer. Paradoxically, sunburns tend to be more frequent among sunscreen users, probably because of greater natural sun sensitivity. When sun exposure was not intentional, sunscreen use did not increase time spent in the sun. Two European double-blind randomized trials conducted among young sun seekers found daily sun exposure duration, especially sunbathing, 19-25% longer with use of SPF 30 than with use of SPF 10 sunscreens. One randomized trial in a holiday resort in France found a 3-13% increase in sun exposure duration with use of SPF 12 versus SPF 40 sunscreen. But, the SPF 12 groups used 3.6-4.2 more sunscreen than the SPF 40 group, and thus the actual SPF in the SPF 12 group was higher than in the SPF 40 groups. In conclusion, sunscreen use leads to longer duration of sun exposure when sun exposure is intentional, but not when sun exposure is non intentional. PMID- 17415717 TI - Improving respiration in patients with tetraplegia by functional electrical stimulation: an anatomical perspective. AB - Patients with tetraplegia often have respiratory complications because of paralysis of the abdominal and intercostal muscles. Functional electrical stimulation (FES) has been used to improve breathing in these patients by applying surface stimulation to the abdominal muscles. We aimed to find the best nerves to stimulate directly to increase tidal volume and make cough more effective. Surface electrodes were placed on a patient's abdominal wall to find the optimum points for surface stimulation. These positions were plotted on a transparent sheet. The abdomino-intercostal nerves were dissected in five male dissecting room cadavers matched for size with the patient. The plastic sheet was then superimposed over each of the dissections to clarify the relationship between optimum surface stimulation points and the underlying nerves. Results show that the optimum surface stimulation points overlie the course of abdomino intercostal nerves T9, 10, and 11. The success with selecting stimulation points associated with T9, 10, and 11 is probably because of the large mass of abdominal muscle supplied by these nerves. The constant position of the nerves below the ribs makes the intercostal space a possible site for direct stimulation of the abdomino-intercostal nerves. PMID- 17415718 TI - Variant superficial fibular (peroneal) nerve anatomy in the middle third of the lateral leg. PMID- 17415719 TI - A variation of fibularis quartus muscles: musculus fibulocalcaneus externum. PMID- 17415720 TI - Central venous catheterization -- an anatomical review of a clinical skill -- Part 1: subclavian vein via the infraclavicular approach. AB - The safe and successful performance of a central venous catheterization (CVC) requires a specific knowledge of anatomy in addition to a working knowledge. Misunderstanding the anatomy may result in failure or complications. This review aims to aid understanding of the anatomical framework, pitfalls, and complications of CVC of the subclavian (SCV). CVC is common practice amongst surgeons, anesthesiologists, and emergency room physicians during the preparations for major surgical procedures such as open-heart surgery, as well as, for intensive care monitoring and rapid restoration of blood volume. Associated with this technique are certain anatomical pitfalls and complications that can be successfully avoided if one possesses a thorough knowledge of the contraindications, regional anatomy, and rationale of the technique. PMID- 17415721 TI - Intra-articular inclusions of the elbow joint complex. AB - An abundant clinical literature implicates intra-articular inclusions in various pathological conditions of the elbow, but the anatomical literature offers piecemeal descriptions of these structures. The aim of this study was to determine the prevalence, disposition, and gross morphology of intra-articular inclusions of the entire elbow joint complex. Twenty-eight elbow joints from 17 adult embalmed cadavera were studied by macrodissection. Three types of inclusions were identified. Fat pads and fibroadipose meniscoids were found in all joints while capsular rims occurred in 50% of radiohumeral and humeroulnar joints. Fat pads were located opposite the olecranon, coronoid, and radial fossae. Fibroadipose meniscoids were found in the nonarticular waists of the trochlear notch and protruded into the posterolateral aspect of the radiohumeral joint. Fibroadipose meniscoids and fat pads are characteristic structures of the elbow joint complex. Fat pads appear to act as deformable space fillers, while the radio-humeral fibroadipose meniscoid appears to protect articular cartilage exposed during normal movement. Arguments are raised whether these structures may act as the nidus for arthrofibrosis and that the radiohumeral meniscoid may be involved in lateral epicondylalgia. PMID- 17415722 TI - Different dental care setting: does income matter? AB - In this paper we consider the use of mobile dental clinics as a means of improving access to dental care among primary school children in Southern Thailand by reducing the opportunity cost of service use to parents. Parents' willingness to pay (WTP) is measured for three different services provided in a community hospital dental clinic and a school-based mobile clinic. Although the service setting does not affect significantly the WTP for treatment directly, the estimated positive association between WTP and income is modified by setting. The results indicate that the potential for mobile clinics to increase utilization of services among primary school children is associated with parents' income, with the difference in valuation of dental services between the two settings being less among lower income parents than higher income parents. However, even among lower income parents our results indicate that the potential for increasing service utilization among children depends on the improvements in access associated with the mobile clinic not being achieved at the opportunity cost of lower levels of effectiveness. PMID- 17415723 TI - Triazolediylidenes: a versatile class of ligands for the preparation of discrete molecules of homo- and hetero-binuclear complexes for improved catalytic applications. PMID- 17415724 TI - Chemically immobilized single-stranded oligonucleotides on praseodymium oxide nanoparticles as an unlabeled DNA sensor probe using impedance. PMID- 17415725 TI - Solvent effects on hydrogen bonding. PMID- 17415726 TI - The mechanism of RNA strand scission: an experimental measure of the Bronsted coefficient, beta nuc. PMID- 17415727 TI - Kinetically stable, flat-lying thiolate monolayers. PMID- 17415728 TI - Enantioselective synthesis of 1,2-diarylaziridines by the organocatalytic reductive amination of alpha-chloroketones. PMID- 17415729 TI - CH3ReO3 on gamma-Al2O3: understanding its structure, initiation, and reactivity in olefin metathesis. PMID- 17415730 TI - Chiral N-heterocyclic carbenes in natural product synthesis: application of Ru catalyzed asymmetric ring-opening/cross-metathesis and Cu-catalyzed allylic alkylation to total synthesis of baconipyrone C. PMID- 17415731 TI - Synthesis of the otteliones A and B: use of a cyclopropyl group as both a steric shield and a vinyl equivalent. PMID- 17415732 TI - Surface-chemistry control to silence gene expression in Drosophila Schneider 2 cells through RNA interference. PMID- 17415733 TI - Directed assembly of cluster-based supramolecules into one-dimensional coordination polymers. PMID- 17415734 TI - Hybrid silica nanoparticles for multimodal imaging. PMID- 17415735 TI - Concise total synthesis of (-)-calycanthine, (+)-chimonanthine, and (+) folicanthine. PMID- 17415736 TI - Large-scale, hot-filament-assisted synthesis of tungsten oxide and related transition metal oxide nanowires. AB - A scalable and versatile method for the large-scale synthesis of tungsten trioxide nanowires and their arrays on a variety of substrates, including amorphous quartz and fluorinated tin oxide, is reported. The synthesis involves the chemical-vapor transport of metal oxide vapor-phase species using air or oxygen flow over hot filaments onto substrates kept at a distance. The results show that the density of the nanowires can be varied from 10(6)-10(10) cm(-2) by varying the substrate temperature. The diameter of the nanowires ranges from 100 20 nm. The results also show that variations in oxygen flow and substrate temperature affect the nanowire morphology from straight to bundled to branched nanowires. A thermodynamic model is proposed to show that the condensation of WO(2) species primarily accounts for the nucleation and subsequent growth of the nanowires, which supports the hypothesis that the nucleation of nanowires occurs through condensation of suboxide WO(2) vapor-phase species. This is in contrast to the expected WO(3) vapor-phase species condensation into WO(3) solid phase for nanoparticle formation. The as-synthesized nanowires are shown to form stable dispersions compared to nanoparticles in various organic and inorganic solvents. PMID- 17415737 TI - Low-temperature synthesis of gamma-alumina nanocrystals from aluminum acetylacetonate in nonaqueous media. PMID- 17415738 TI - New light on an old story: formation of melam during thermal condensation of melamine. AB - We report on the existence and formation of the carbon nitride precursor melam (H(2)N)(2)(C(3)N(3))NH(C(3)N(3))(NH(2))(2), thereby clarifying one of the last unresolved issues posed by the complex thermal condensation of melamine C(3)N(3)(NH(2))(3). Experimental proof is put forward that melam is a direct condensation product of melamine, but can be detected only in small amounts under special reaction conditions owing to its rapid transformation into melem. The coexistence of melamine and melem during thermal condensation yields two adduct phases with distinct compositions [C(3)N(3)(NH(2))(3)](2)[C(6)N(7)(NH(2))(3)] and [C(3)N(3)(NH(2))(3)][C(6)N(7)(NH(2))(3)](2). They may be considered as co crystallizates of melamine and melem and can be isolated as intermediates between 590 and 650 K prior to the formation of single-phase melem C(6)N(7)(NH(2))(3). Melam (C2/c, a=1811.0(4), b=1086.7(2), c=1398.4(3) pm, beta=96.31(3) degrees, V=2735.3(9)x10(6) pm(3), T=130 K) adopts a ditriazinylamine-type structure with a twisted conformation about the bridging NH moiety and transforms into melem around 640 K. Two compounds deriving from melam have been synthesized by solution and solid-state reactions. The salt melamium diperchlorate C(6)N(11)H(11)(ClO(4))(2).2H(2)O (C2/c, a=1747.8(4), b=1148.2(2), c=993.6(2) pm, beta=118.79(3) degrees, V=1747.4(6)x10(6) pm(3), T=130 K) crystallizes as a dihydrate and exhibits a doubly protonated, planar melam core. In the neutral complex Zn[C(6)N(11)H(9)]Cl(2) (P2(1)/c, a=743.00(15), b=2233.2(5), c=762.5(2) pm, beta=99.86(3) degrees, V=1246.5(4)x10(6) pm(3), T=200 K), melam acts as a symmetrically bent bidentate ligand, which is coordinated to the Lewis acid Zn site through two ring nitrogen atoms. PMID- 17415739 TI - Unmasking melon by a complementary approach employing electron diffraction, solid state NMR spectroscopy, and theoretical calculations-structural characterization of a carbon nitride polymer. AB - Poly(aminoimino)heptazine, otherwise known as Liebig's melon, whose composition and structure has been subject to multitudinous speculations, was synthesized from melamine at 630 degrees C under the pressure of ammonia. Electron diffraction, solid-state NMR spectroscopy, and theoretical calculations revealed that the nanocrystalline material exhibits domains well-ordered in two dimensions, thereby allowing the structure solution in projection by electron diffraction. Melon ([C(6)N(7)(NH(2))(NH)](n), plane group p2 gg, a=16.7, b=12.4 A, gamma=90 degrees, Z=4), is composed of layers made up from infinite 1D chains of NH-bridged melem (C(6)N(7)(NH(2))(3)) monomers. The strands adopt a zigzag type geometry and are tightly linked by hydrogen bonds to give a 2D planar array. The inter-layer distance was determined to be 3.2 A from X-ray powder diffraction. The presence of heptazine building blocks, as well as NH and NH(2) groups was confirmed by (13)C and (15)N solid-state NMR spectroscopy using (15)N labeled melon. The degree of condensation of the heptazine core was further substantiated by a (15)N direct excitation measurement. Magnetization exchange observed between all (15)N nuclei using a fp-RFDR experiment, together with the CP-MAS data and elemental analysis, suggests that the sample is mainly homogeneous in terms of its basic composition and molecular building blocks. Semiempirical, force field, and DFT/plane wave calculations under periodic boundary conditions corroborate the structure model obtained by electron diffraction. The overall planarity of the layers is confirmed and a good agreement is obtained between the experimental and calculated NMR chemical shift parameters. The polymeric character and thermal stability of melon might render this polymer a pre-stage of g-C(3)N(4) and portend its use as a promising inert material for a variety of applications in materials and surface science. PMID- 17415740 TI - Anion-templated calix[4]arene-based pseudorotaxanes and catenanes. AB - We present the rational design and anion-binding properties of the first anion templated pseudorotaxanes and catenanes in which the "wheel" component is provided by a calix[4]arene macrobicyclic unit. The designs and syntheses of two new calix[4]arene macrobicycles, 2 and 3, are presented, and the abilities of these new species both to bind anions and to undergo anion-dependent pseudorotaxane formation are demonstrated. Furthermore, it is shown that performing ring-closing metathesis reactions on some of these pseudorotaxane assemblies gives novel catenane species 14 and 15, in which the yield of interlocked molecule obtained is critically dependent on the presence of a suitable anion template, namely, chloride. Exchange of the chloride anion in catenane 14 a for hexafluorophosphate gives catenane 14 d, which contains a unique anion-binding domain defined by the permanently interlocked hydrogen-bond donating calix[4]arene macrobicycle and pyridinium macrocycle fragments. The anion-binding properties of this domain are presented, and shown to differ from non-interlocked components. PMID- 17415741 TI - Beyond the structure-property relationship paradigm: influence of the crystal structure and microstructure on the Li+ conductivity of La2/3Li(x)Ti(1-x)Al(x)O3 Oxides. AB - The crystal structures of several oxides of the La(2/3)Li(x)Ti(1-x)Al(x)O(3) system have been studied by selected-area electron diffraction, high-resolution transmission electron microscopy, and powder neutron diffraction, and their lithium conductivity has been by complex impedance spectroscopy. The compounds have a perovskite-related structure with a unit cell radical2 a(p)x2 a(p)x radical2 a(p) (a(p)=perovskite lattice parameter) due to the tilting of the (Ti/Al)O(6) octahedra and the ordering of lanthanum and lithium ions and vacancies along the 2 a(p) axis. The Li(+) ions present a distorted square-planar coordination and are located in interstitial positions of the structure, which could explain the very high ionic conductivity of this type of material. The lithium conductivity depends on the oxide composition and its crystal microstructure, which varies with the thermal treatment of the sample. The microstructure of these titanates is complex due to formation of domains of ordering and other defects such as strains and compositional fluctuations. PMID- 17415742 TI - Bone formation as a potential motive force of tooth eruption in the rat molar. AB - The objectives of this anatomical study were to (1) determine if significant bone growth occurs in the base of the alveolar bony crypt of the first mandibular molar to move the tooth through the eruption pathway; (2) determine if the osteogenesis in the crypt correlates with the published chronological gene expression of bone morphogenetic protein-2 (BMP-2) in the dental follicle; and (3) determine chronologically and regionally the crypt bone activity. To accomplish this, the alveolar bony crypts of rat mandibular molars from postnatal days 3 to 18 were processed and examined by scanning electron microscopy (SEM). In addition, mandibles and teeth of ages 12-18 were prepared for light microscopy. SEM demonstrated that bone formation occurs in the basal (apical) portion of the alveolar bony crypt at day 3, whereas bone resorption concurrently is ongoing in the coronal region of the crypt. By day 9, the crypt is beginning to be reduced in depth as the result of basal bone formation, and by day 14, the base of the crypt immediately under the tooth is almost completely filled with bone to form the interradicular septum. At day 18, the tooth erupts as bone formation likely elevates the molar. Bone growth in the basal area of the crypt correlates with a previous study showing enhanced BMP-2 expression in the dental follicle. Thus, SEM indicates that the motive force of tooth eruption likely is bone formation at the base of the alveolar crypt and this osteogenesis may relate to BMP-2 production in the dental follicle. PMID- 17415743 TI - The basilar venous plexus. AB - The basilar venous plexus is the anteromedian venous channel of the posterior cranial fossa that has many conflicting and brief descriptions in the extant literature. To our knowledge, no single study has been performed that analyzed this venous structure in detail. The aim of the current study was to elucidate further the anatomy of this structure of the posterior cranial fossa. The authors examined twenty adult cadaveric specimens following injection of the internal jugular veins or cavernous sinus to observe the morphology of the basilar plexus. All specimens were found to have a basilar plexus which was always plexiform and very variable in nature. This structure was dorsal to the clivus superiorly and dorsal clivus and overlying tectorial membrane inferiorly. The mean diameter of the channels making up this plexus was 1.1 mm. Communication was always found between the basilar plexus and the inferior petrosal sinuses and this was the primary route used to drain the basilar sinus out of the cranium. In fact, these two venous structures were more or less contiguous across the midline at multiple levels. In seven specimens (35%), the basilar plexus communicated with veins draining into the hypoglossal canal. The basilar plexus communicated with the marginal sinus in 12 specimens (60%). This plexus became much less concentrated as it descended inferior to a plane between the jugular tubercles. No specimen was found to have connections with veins of the anterior brain stem or ventral surface of the clivus. The basilar plexus is a highly variable posterior fossa venous structure. Clinicians and radiologists should take into account this variability when managing cerebral venous disorders or interpreting imaging studies of the skull base. PMID- 17415744 TI - Thoracic radicular vessels by simultaneous intra-arterial and intravenous injection of araldite. AB - Spinal dural arteriovenous fistulas (SDAVF) consist of a shunt between a radicular artery and a radicular vein, resulting in a progressive paraparesis. They are most prevalent in middle-aged men (male to female ratio 5 to 1). It is unknown why the shunt develops. It is possible that there are anatomical differences between men and women, which may account for the sex difference in prevalence. We performed a study with simultaneous arterial and venous araldite injection in 5 male and 5 female human cadavers using different colors. The mean age of the human cadavers was 78 years (range 70-91). The human cadavers were not known to have suffered from spinal disease. We did not find significant differences in thoracic vasculature between men and women. Two different types of radicular arteries could be identified: The first was the arterial feeder of the root ganglion or the dura mater, which is also called the distal radicular artery. The second was the tributary of the anterior spinal artery, which is also called the medullary artery. We found three arteriovenous anastomoses between the radicular artery and the corresponding vein, and three between the radicular artery and venous plexus. We found a total of six thoracic arteriovenous shunts in four cadavers but their role in the pathogenesis of SDAVF remains uncertain. No vascular anatomic differences between men and women were found. PMID- 17415745 TI - An accessory muscle within the suboccipital triangle. PMID- 17415746 TI - Cisterna chyli: a detailed anatomic investigation. AB - With recent laparoscopic advancements in retroperitoneal and thoracic surgical procedures, familiarity with major lymphatic structures, such as including the cisterna chyli (CC) and thoracic duct (TD), has proven beneficial in avoiding misdiagnosis and iatrogenic intraoperative injury. In this light, the aim of our study was to explore and delineate the topography of the CC, classify the different patterns of lymphatic tributaries, and categorize its varying location with respect to the vertebral bodies. The anatomy of the CC was examined in 120 adult human cadavers. The CC was found in 83.3% of the specimens and both the tributaries of the CC and the location, with respect to vertebral level, demonstrated wide variation. The results were classified into four types. The most common tributary configuration (type I), found in 45% specimens, was a single CC formed by the union of the left lumbar trunk (LT) and the intestinal trunk (IT). In 30% the CC was formed where the IT opened into the TD and the right lumbar trunk (RT), LT, retroaortic nodes (RN) and branches from the intercostal lymphatics (IL) joined variably (type II). In 20% the CC was formed by the junction of the RT and IT (type III), while in 5% there was a variable confluence pattern of lymphatic trunks that could not be classified (type IV). The CC was located at L1-L2 (type A) in 63%, T12-L1 (type B) in 21%, T11-T12 (type C) in 8%, T10-11 (type D) in 5%, and T9-10 (Type E) in 3%, of the specimens. The CC was found in the retrocrural space and, in 75% of the cases, to the right of the abdominal aorta. We hope that the data supplied by this study will provide useful information in the future to anatomists, radiologists and surgeons alike. PMID- 17415747 TI - Gliclazide: pharmacokinetic-pharmacodynamic relationships in rats. AB - The relationship between the pharmacokinetics of gliclazide and its antidiabetic efficacy were evaluated on the basis of experimental determination of changes with time in the plasma levels of this antidiabetic agent and those of glucose. The experiment included rats with both initial normal glycaemia and alloxan induced hyperglycaemia (glycaemia increased by a minimum of 30%). Pharmacokinetic and pharmacodynamic parameters were examined in the interval of 30 to 180 min after p.o. administration of a single dose of 25 mg/kg of gliclazide. The drug was administered on day 4, following a single i.v. dose of either 50 mg/kg of alloxan (hyperglycaemic group) or the injection vehicle (control group). Even though the biological availability of gliclazide was similar in both normoglycaemic and hyperglycaemic animals, the gliclazide-induced hypoglycaemizing response was not uniform: until 60 min, the decrease of glycaemia was smaller in animals with alloxan hyperglycaemia (23% decrease at 60 min) in contrast to the normoglycaemic animals (36% decrease at 60 min), at later times, the intensity of this hypoglycaemizing effect of gliclazide persisted in the hyperglycaemic animals, while in the normoglycaemic ones, a reversal of the hypoglycaemizing effect occurred. PMID- 17415748 TI - Environmental complexity affects contextual fear conditioning following hippocampal lesions in rats. AB - Contextual fear conditioning has become a benchmark measure for hippocampal function, even though several studies report successful acquisition in hippocampal-damaged rodents. The current study examined whether environmental complexity may account for these discrepancies. We directly compared single session contextual fear conditioning in rats in a simple vs. complex environment. Hippocampal lesions led to reduced fear conditioning in both contexts, as measured by freezing, but the effect was significantly greater in the complex context. As well, lesions led to generalized fear when the complex context was paired with shock, but not when the simple context was paired. We suggest that the representation of the simple context formed by rats with hippocampal lesions was adequate to support associative learning, but the representation of the complex context, which depended to a greater extent on relational learning, was not. The results were interpreted as consistent with theories of hippocampal function that emphasize its role in integrating multiple stimulus elements in a memory trace. PMID- 17415749 TI - Retroperitoneal hemorrhage from inferior epigastric artery. PMID- 17415750 TI - The sural nerve is not always a pure sensory nerve. PMID- 17415751 TI - The sperm nuclear matrix is required for paternal DNA replication. AB - The mammalian sperm nucleus provides an excellent model for studying the relationship between the formation of nuclear structure and the initiation of DNA replication. We previously demonstrated that mammalian sperm nuclei contain a nuclear matrix that organizes the DNA into loop domains in a manner similar to that of somatic cells. In this study, we tested the minimal components of the sperm nucleus that are necessary for the formation of the male pronucleus and for the initiation of DNA synthesis. We extracted mouse sperm nuclei with high salt and dithiothreitol to remove the protamines in order to form nuclear halos. These were then treated with either restriction endonucleases to release the DNA not directly associated with the nuclear matrix or with DNAse I to digest all the DNA. The treated sperm nuclei were injected into oocytes, and the paternal pronuclear formation and DNA synthesis was monitored. We found that restriction digested sperm nuclear halos were capable of forming paternal pronuclei and initiating DNA synthesis. However, when isolated mouse sperm DNA or sperm DNA reconstituted with the nuclear matrices were injected into oocytes, no paternal pronuclear formation or DNA synthesis was observed. These data suggest that the in situ nuclear matrix attachment organization of sperm DNA is required for mouse paternal pronuclear DNA synthesis. PMID- 17415752 TI - Longitudinal excursion and strain in the median nerve during novel nerve gliding exercises for carpal tunnel syndrome. AB - Nerve and tendon gliding exercises are advocated in the conservative and postoperative management of carpal tunnel syndrome (CTS). However, traditionally advocated exercises elongate the nerve bedding substantially, which may induce a potentially deleterious strain in the median nerve with the risk of symptom exacerbation in some patients and reduced benefits from nerve gliding. This study aimed to evaluate various nerve gliding exercises, including novel techniques that aim to slide the nerve through the carpal tunnel while minimizing strain ("sliding techniques"). With these sliding techniques, it is assumed that an increase in nerve strain due to nerve bed elongation at one joint (e.g., wrist extension) is simultaneously counterbalanced by a decrease in nerve bed length at an adjacent joint (e.g., elbow flexion). Excursion and strain in the median nerve at the wrist were measured with a digital calliper and miniature strain gauge in six human cadavers during six mobilization techniques. The sliding technique resulted in an excursion of 12.4 mm, which was 30% larger than any other technique (p/= 18 years with histologically proven recurrent or metastatic nasopharyngeal carcinoma, an Eastern Cooperative Oncology Group (ECOG) performance status /= 12 weeks were eligible. Measurable disease outside prior radiotherapy ports was required, unless a subsequent progression of the lesion was documented. An interval of >/= 12 months was required between the previous chemotherapy (neoadjuvant, concurrent chemoradiotherapy or adjuvant) and study entry. Prior radiotherapy or surgery were allowed. All patients had adequate bone marrow (WBC >4000/mL, platelets >100000/mL), hepatic (bilirubin <1,5 mg/dL, SGPT <1.5xN), and renal function (serum creatinine <1.5 mg/dL or creatinine clearance >60 mL/min). Chemotherapy consisted of paclitaxel 175 mg/m(2), given as a 3-hour infusion, followed by carboplatin dosed to an area under the concentration- time curve (AUC) of 6 mg*min/mL, administered every 21 days. RESULTS: 40 patients entered the study. There were 3 complete responders (CR) and 8 partial responders (PR), for an overall response rate (ORR) of 27.5% (95% confidence interval - C.I.: 14.5-44). Median time to progression (TTP) was 3.5 months, and median survival was 11.5 months. Grade 3-4 toxicity included leucopenia (17.5% of the patients), anaemia (17.5%), thrombocytopenia (10%), neutropenia (7.5%), and peripheral neuropathy (2.5%). CONCLUSION: These data indicate that the combination of paclitaxel and carboplatin can be safely administered on an outpatient basis, but it is only moderately active against relapsed/metastatic nasopharyngeal carcinoma patients. PMID- 17415809 TI - Split course radiation with concurrent vinorelbine and cisplatin in locally advanced non-small cell lung cancer. A phase II study. AB - PURPOSE: Recent results coming from large randomized trials suggest that for locally advanced non-small cell lung cancer (NSCLC), integration of chemotherapy (CT) with irradiation (RT) should be concurrent rather than sequential. This study aimed at evaluating the actually delivered RT and CT dose intensities (DI), along with the toxicity and efficacy of a split course RT program with concurrent CT. PATIENTS AND METHODS: From October 2000 to September 2002, 24 patients with histologically or cytologically documented NSCLC were included. Patients' characteristics were as follows: males/females=22/2, median age=59 years, stage IIIB/IIIA=22/2 patients, ECOG PS 0-1=15 (62%) and PS 2=9 (38%). HISTOLOGY: adenocarcinoma/ squamous cell/large cell/unclassified 10/6/1/7, respectively. Four cycles of vinorelbine (VNB) 25 mg/m(2) and cisplatin (CDDP) 40 mg/m(2) on days 1+8 were administered (days 1,8,22,29,57,64,78,85). Concurrent with the second CT cycle, RT (2 courses of 30 Gy separated by a 2-week break) was delivered, with a plan to achieve a total dose of 60 Gy, with a fractionation schedule of 2 Gy/day/5 days weekly. RESULTS: The intended RT dose was delivered to 21 (88%) patients with a relative DI of 0.93. Nineteen (79%) patients received more than 3 CT cycles. The relative DI for VNB and CDDP were 0.88 and 0.83, respectively. During treatment 3 (13%) patients experienced WHO grade 3-4 hematologic toxicity while ECOG grade 3 esophagitis was recorded also in 3 patients. At the end of treatment 14 (58%) patients achieved an objective response (2 complete - CR and 12 partial response - PR), while 8 (33%) patients had stable disease (SD) and 2 (8%) progressive disease (PD). After a median follow up of 15 months (range 3-26), 15 (62%) patients relapsed. There were 8 (33%) patients with local relapse and 7 (29%) with distant metastases. The median progression free (PFS) and overall survival (OS) were 10 (range 2-24) and 15 (range 5-24) months, respectively, with an estimated 1 and 2-year survival rates of 55% and 10%, respectively. CONCLUSION: Our concurrent schedule allows for good CT and RT DI, with low associated toxicities. The efficacy data are considered promising, taking into account the high proportion of stage IIIB patients evaluated. PMID- 17415810 TI - Correlation of steroid hormone receptor status with histological and nuclear grading in breast carcinoma. AB - PURPOSE: The present study was performed to evaluate the immunohistochemical analysis of estrogen receptor (ER) and progesterone receptor (PR) in invasive breast carcinomas of various histological subtypes and grades. In this paper an attempt was made to establish a correlation between hormone receptor status and histological and nuclear grading of breast carcinoma. MATERIALS AND METHODS: Immunohistochemistry was performed on paraffin sections of 80 invasive breast carcinomas (38 ductal, 18 lobular, 18 ducto-lobular, 2 medullary, 2 mucinous, 1 tubular and 1 papillary). The same scoring system was used for immunohistochemically stained ER and PR. The results were compared with the histological and nuclear grade and analyzed by the chisquare test. RESULTS: Positive immunoreactivity for ER and PR were seen in 71.25% and 60.00% cases, respectively. Both ER and PR positive immunostaining was observed in all (100%) well-differentiated (grade I) breast carcinomas, while in grade II tumors ER and PR-positive cancer cells were 76.36% and 61.62%, respectively. The corresponding figures for grade III carcinomas were 41.18% and 35.29%. A significant association (p <0.05) between different histological grades of breast carcinomas and ER and PR immunoreactivity was found. No significant association was found between nuclear grade of breast carcinoma and ER and PR immunoreactivity. CONCLUSION: The results presented herein suggest that histological grade of invasive breast carcinoma was significantly associated with ER and PR immunoreactivity, while nuclear grade alone showed no correlation. Moreover, our findings showed that ER and PR positivity declined with increasing tumor grade. PMID- 17415811 TI - Do we really benefit from checking tumor markers in detecting recurrence in gastrointestinal cancer? AB - PURPOSE: To evaluate the role of tumor markers carcinoembryonic antigen (CEA) and CA 19.9 in the early detection of local or systemic recurrence in gastrointestinal malignancies. PATIENTS AND METHODS: Twenty-six patients with operable gastrointestinal cancer, who had elevated levels of either CEA or CA 19.9 or both during the postoperative follow-up period were evaluated. Serum estimation of tumor markers were carried out at 3-month intervals and the imaging and endoscopic procedures were performed at 6-month intervals or when a patient had an elevated tumor marker during follow-up. RESULTS: The difference of mean serum levels of CA 19.9 but not of CEA was found to be statistically significant between the two groups of patients with or without radiographically / endoscopically evident recurrent disease (p < 0.05). CONCLUSION: CA 19.9 was found to be a better, though not specific, indicator of recurrence. The relative small number of patients precludes reaching a firm conclusion. Further studies are needed to establish the role of these markers in determining early recurrence and their impact in overall survival. PMID- 17415812 TI - Immunohistochemical analysis of prostate-specific antigen in female breast cancer. AB - PURPOSE: Prostate-specific antigen (PSA), a glycoprotein initially thought to be produced only by the epithelial cells of the prostate, has recently been found in various tissues and tumors. It has been suggested that the expression of PSA in breast cancer is a good prognostic indicator and correlated with favorable prognosis. However, in recent years opposite results have been reported. In this study, we investigated the immunoreactivity of PSA in female breast cancer to find out any relationship between PSA and prognostic parameters. PATIENTS AND METHODS: Sections of formalin-fixed, paraffin- embedded samples from 109 invasive ductal carcinomas were immunostained for PSA. The staining results were analyzed in relation to age, tumor size, histologic grade, axillary lymph node status and steroid receptors. RESULTS: PSA immunoreactivity was seen in only 11 (10.1%) out of 109 cases. All PSA positive cases were also estrogen (ER) and progesterone receptor (PR) positive. We found a statistically significant correlation between PSA and the expression of steroid receptors, while no correlation was detected with the other factors. CONCLUSION: The detection of PSA, using immunohistochemistry, does not seem to be a reliable prognostic criterion for female breast cancer patients or a marker of tumor origin. PMID- 17415813 TI - The role of ascorbic acid, selenium, and glutathione on benzo[a]pyrene-induced carcinogenesis in wistar rats. AB - PURPOSE: The carcinogenic action of polycyclic aromatic hydrocarbons (PAHs) can be inhibited by endogenous or exogenous compounds. This study was designed to elucidate the modifying action of 3 endogenous inhibitors- ascorbic acid (vit C) used alone, and selenium (Se) used in combination with glutathione (GSH). MATERIALS AND METHODS: Chemical carcinogenesis was induced by benzo[a]pyrene(BaP). A hundred wistar rats were divided into 3 groups: the first group (G I) consisted of 42 animals, representing the control group. The two experimental groups (G II and G III) consisted of 38 and 20 rats, respectively. All groups were injected with BaP(10.08 mg subcutaneously-s.c). The first experimental G II was given only vit C (520 mg in 2% sugar solution per os - p.o.). The second experimental G III was given Se (0.1 mg p.o.) with GSH (200 mg p.o.). Tumor incidence and mean survival time were determined. Histological examination of the developed and excised tumors took place following death. The carcinogenic potency (CP) and anticarcinogenic potency (AP) of the substances used were calculated. RESULTS: A statistically significant difference regarding the mean survival time in the two experimental groups (238.4-/+31 days and 344.9 /+48 days, respectively) compared to the control group (183.8-/+28 days) was found (p < 0.001). The CP of each of the 3 groups was 54.3, 41.2, and 28.9 units, respectively. The AP of vit C used alone was 13.1 units, representing a significant anticarcinogenic effect. The combination of Se + GSH showed an AP of 25.4 units, resulting in a significant prolongation of the mean survival time, which is considered a potent anticarcinogenic effect. Furthermore, a statistically significant difference was found also when the mean survival time of G III animals was compared with G II. CONCLUSION: Vit C on its own and Se in combination with GSH represent strong endogenous inhibitors that can inhibit/reduce the carcinogenic action of BaP-induced carcinogenesis in wistar rats. The combination therapy used offered better in vivo results. PMID- 17415814 TI - Do collimator angle setup errors cause dose inhomogeneity at the junction zone during craniospinal irradiation? A phantom study. AB - PURPOSE: Craniospinal irradiation (CSI) has some geometric uncertainties, especially at the junction zone. In this study we tried to evaluate how possible random setup errors of the collimator angle may contribute to these uncertainties. MATERIALS AND METHODS: Cranial and spinal fields were drawn on RW3 solid water phantom in accordance with the divergence matching technique (DMT). Field dimensions were 18x18 cm and 6x30 cm, respectively. We placed light insulated Kodak X-Omat V films at the junction zone, then we irradiated the films with different collimator angles with both Co-60 and 4 MV conditions, and determined how the junction zone was affected from random setup errors in DMT. RESULTS: 10.6 degrees collimator angle was proper for 30 cm upper spinal field. For Co-60 machine the dose homogeneity of this angle was +4.5%. For the angles of 8, 9, 11 and 12 degrees the homogeneities were -13%, - 11%, +5% and +10 %, respectively. For 4 MV photon the dose homogeneity of the 10.6 degrees collimator angle was +3%. For the angles of 8, 9, 11 and 12 degrees the homogeneities were 17%, -14%, +6 % and +13%, respectively. CONCLUSION: As the CSI has some geometric uncertainties, serious dose inhomogeneities may occur at the junction zone. The collimator angle is of great importance and any random setup errors may not be tolerated. PMID- 17415815 TI - Clonal heterogeneity in a patient with acute promyelocytic leukemia. AB - A 18-year-old man was diagnosed with acute promyelocytic leukemia (APL). The conventional cytogenetic analysis revealed normal karyotype 46, XY, t(15; 17). Reverse transcriptase polymerase chain reaction (RTPCR) identified PML-RARa chimeric transcripts. Complete remission (CR) was attained with 3 induction courses of Ara-C, daunorubicin and all-trans retinoic acid (ATRA). Three years later the patient relapsed. The blasts in bone marrow aspirate at relapse had AML M3 morphology, and RT-PCR was positive for PML-RARa transcripts. The patient was treated with ATRA and daunorubicin without success. Two months later the blasts in bone marrow aspirate showed AML-M2 morphology, the karyotype was 47, XY, +8 and RT-PCR revealed the presence of AML1-ETO transcripts and absence of PML-RARa transcripts. The patient attained second CR with 3 induction courses -a course with Ara-C and daunorubicin and 2 courses with idarubicin, Ara-C and etoposide. PMID- 17415816 TI - Unusual metastasis of renal cell carcinoma to the nasal cavity. AB - Metastasis of renal cell carcinoma (RCC) to the nose and paranasal sinuses is very rare. We present two cases of RCC metastatic to the nose who were treated with palliative radiotherapy. Although the prognosis was poor for both of the patients, the main symptoms (epistaxis and unilateral airway obstruction) were successfully palliated with irradiation. PMID- 17415817 TI - Sclerosing stromal tumor of the ovary: report of three cases. AB - We present 3 cases displaying sclerosing stromal tumor (SST) of the ovary and describe their immunohistochemical and electron microscopic findings. The patients were 23, 24 and 28-year-old and had an ovarian mass. Histologically, the tumors had the typical appearance of SST of the ovary. A pseudolobular pattern which was composed of cellular areas and less cellular areas was seen on low power microscopy. PMID- 17415818 TI - A case of nevoid basal cell cutaneous syndrome - radiological, CT and MRI findings. AB - The nevoid basal cell carcinoma syndrome (NBCCS) is a rare, still well-defined autosomal dominant disorder, characterized by multiple cutaneous basal cell carcinomas, jaw keratocysts and a variety of other tumors and developmental anomalies. The nevi turn malignant with time, and thus, early diagnosis, follow up, and treatment are imperative. This article presents a case of a 60-year-old man with classical form of NBCCS. The images from conventional radiography, CT and MRI are discussed in the context of clinical presentation of the disease, and the importance of imaging modalities in the diagnostic process is stressed. PMID- 17415819 TI - Theodor Billroth (1829-1894) and other protagonists of gastric surgery for cancer. AB - Theodor Billroth was one of the great surgical giants of all time. In Vienna he created one of the finest schools in the history of surgery where he carried out pioneering work in experimental studies, surgical pathology and operative surgery. He pioneered gastric surgery for cancer. In 1881 he carried out the first successful partial gastrectomy for cancer. The first pylorectomy was carried out in 1879 by Jules Pean of Paris but his patient died on the fifth postoperative day. PMID- 17415821 TI - Colorectal cancer and chemoprevention. PMID- 17415820 TI - Basics of cancer pharmacogenomics. PMID- 17415822 TI - The significance of perioperative anemia in patients with resectable gastrointestinal tract tumors. AB - Cancer-related anemia occurs in more than 50% of patients with malignancies and constitutes a common finding in patients with gastrointestinal tract tumors. In the present article we present the possible pathogenetic mechanisms as well as the appropriate clinical and laboratory investigations for the anemia, which is associated with gastrointestinal tract malignancies. Moreover, we conducted a MEDLINE database search between 1992- 2003, focusing on the currently available methods for perioperative correction and treatment of anemia. The currently available evidence suggests that perioperative allogenic blood transfusion is associated with increased rates of postoperative infections and constitutes an independent adverse prognostic factor in gastrointestinal malignancies; autologous blood transfusions are of no clinical benefit compared to allogenic transfusions, as autologous blood is not immunologically neutral; and the shortterm results of erythropoietin (EPO) use remain controversial, while its long-term results remain unknown. Correction of anemia in every patient with resectable gastrointestinal malignancy is mandatory, because it improves surgical stress response, wound healing process and quality of life. Although EPO administration constitutes the treatment of choice for patients receiving chemotherapy and/ or radiotherapy, the best perioperative method for anemia correction remains unknown and further prospective randomized studies are required. From the surgical point of view, any effort for "bloodless surgery" should be attempted. PMID- 17415823 TI - Phase I study of postoperative radiotherapy with concomitant weekly irinotecan, 5 fluorouracil and folinic acid in locally advanced rectal cancer. AB - PURPOSE: 5- fluorouracil (5-FU)-based chemotherapy with concomitant pelvic radiotherapy represents the gold standard of the adjuvant treatment of high-risk rectal cancer. This study aimed to determine the maximum tolerated dose (MTD) of weekly irinotecan (CPT-11) when combined with fixed 5FU/FA doses and pelvic irradiation. PATIENTS AND METHODS: Twenty- four patients with stage II or III rectal cancer were accrued. All had undergone curative surgery before entering the study. Standard pelvic radiotherapy was delivered (50.4 Gy, 1.8 Gy/ fraction in 5.5 weeks). The 5-FU/FA doses were 350/250 (mg/m(2)) in the first 6 patients and 250/100 in the remaining patients. Weekly doses of CPT-11 started at 30 mg/m(2) with escalation steps of 10 mg/m(2). CPT-11 was escalated when 3 patients had been monitored for 8 weeks, without a dose limiting toxicity (DLT). RESULTS: Twenty-three out of 24 patients completed the chemoradiation course. Only 1 patient discontinued the treatment due to persistent grade 3 diarrhea. Of the 144 planned weekly chemotherapy cycles, only 7 were omitted as a result of persisting grade 2-3 gastrointestinal toxicity in 3 patients and grade 3 neutropenia in 1 patient. Grade 3 gastrointestinal DLTs were observed at doses at the level of 30/250/100 in 1 patient and 70/250/100 in 2 patients. Late DLTs were severe radiation dermatitis and colitis at 40/ 350/250 (1 patient) and 70/250/100 (2 patients), respectively. With a follow-up of 18 months 20 (83.3%) patients remain disease- free. CONCLUSION: The administration of weekly CPT-11/ 5FU/FA with concomitant pelvic radiotherapy is feasible and effective. This treatment schedule is associated with mild myelosuppression and mild to moderate gastrointestinal toxicity. Caution should be paid on late radiotherapy-induced toxicities. The MTD of weekly CPT-11 is 30 mg/m(2) when combined with 5FU/FA doses (mg/m(2)) of 350sol;250 and reaches 60 mgsol;m(2) with lower doses of 5FU/FA (250/100). PMID- 17415824 TI - A randomized trial of 5-fluorouracil, leucovorin, cisplatin and epirubicin (PELF) versus 5-fluorouracil, leucovorin and etoposide (ELF) given as adjuvant chemotherapy to patients with resected advanced gastric adenocarcinomas. AB - PURPOSE: To compare the efficacy and toxicity between 2 regimens [5 fluorouracil (5-FU), leucovorin and etoposide (ELF) and 5-FU, leucovorin, cisplatin and epirubicin (PELF)] administered as postoperative adjuvant chemotherapy to patients with completely resected advanced gastric cancer. PATIENTS AND METHODS: Between 1998-2002, 78 patients with advanced gastric cancer were randomly assigned to receive 6 cycles of adjuvant ELF or PELF combination chemotherapy after complete surgical tumor resection. Endpoints were disease-free survival (DFS), overall survival and treatment toxicity. RESULTS: ELF was administered to 37 and PELF to 41 patients. Median overall survival was 12.3 months in the ELF group and 17.2 months in the PELF group (p=0.01), respectively. For the ELF group the median DFS was 17 weeks (range 7-160 weeks), while for the PELF group it was 35 weeks (range 12-172 weeks) (p=0.0004). Two-year overall survival was 8% (3 patients) in the ELF group and 24% (10 patients) in the PELF group (p=0.03). Grade 2 hematologic toxicity occurred in 21% (8 patients) in the ELF group and in 22% (9 patients) in the PELF group (p=0.5) and grade 3 in 29% (11 patients) in the ELF group and in 24% (10 patients) in the PELF group (p=0.2). Grade 2 non hematologic toxicity was seen in 8% (3 patients) in the ELF group and in 17% (7 patients) in the PELF group (p=0.2). Grade 3 non-hematologic toxicity occurred in 29% (11 patients) in the ELF group and in 26.8% (11 patients in the PELF group) (p=0.2). CONCLUSION: PELF combination chemotherapy resulted in DFS and overall survival advantage compared with ELF. No significant differences in hematologic or non-hematologic toxicities between the two groups were registered. PELF is superior to ELF and represents a valid option for the treatment of gastric cancer. PMID- 17415825 TI - Radical radiotherapy in the treatment of carcinoma of the anal canal - single center experience. AB - PURPOSE: Radiotherapy -/+ chemotherapy is the first-line treatment for carcinoma of the anal canal in most oncology institutions. The aim of our study was to evaluate the efficacy and toxicity of radical radiotherapy in the treatment of this carcinoma. PATIENTS AND METHODS: Definitive radiotherapy was performed in 41 patients with squamous cell anal carcinoma. The majority (51.2%) of them were in T3 stage. Positive inguinal lymph nodes were found in 10 (24.4%) patients. Initially, all patients received external beam radiotherapy with pelvic fields, followed, after a 2-week gap in some patients or in continuity, by local external beam or brachytherapy boost. The total tumor dose ranged from 55-75 Gy. RESULTS: Acute complications were noticed in 32 (78%) patients with moist skin perineal desquamation being the most frequent (63.5%). On the first follow-up examination (2 months after the end of radiotherapy) 65.9% of the patients were in complete remission (CR), 31.7% in partial remission (PR), while in one (2.4%) patient locoregional disease progression (PD) was registered. With a mean follow- up time of 38.4 months (range 12-90 months) 75.6% of the patients were disease-free, while local or distant disease progression was diagnosed in 24.4% of them. Local disease control was achieved in 82.9% of the patients. The 5-year overall survival was 76.59%. Late sequelae, which were of low grade (1 and 2), were registered in 22 (53.7%) patients. CONCLUSION: Our study confirmed good treatment results and acceptable toxicity of definitive radiotherapy in the treatment of anal canal carcinoma. PMID- 17415826 TI - Antitumour effect of a- and d- lactam androgen nitrogen mustards on non-small cell lung carcinoma. AB - PURPOSE: We tested 3 alkylating esters of D-lactam androsterone, 3 alkylating esters of A-lactam testosterone and the alkylating nitrogen mustard components of these esters, for antineoplastic activity on non-small cell lung carcinoma (NSCLC) in vitro and in vivo. MATERIALS AND METHODS: Cytostatic and cytotoxic activity was evaluated in vitro against 10 human NSCLC cell lines. The in vitro testing was performed with the MTT metabolic-colorimetric assay and the mean concentrations of each drug that generated 50% or total (100%) growth inhibition (GI50 and TGI, respectively) as well as the drug concentrations that produced cytotoxicity against 50% of the cultured cells (IC50) were calculated. Furthermore, the in vivo antitumour effect was determined against the relatively chemo-resistant Lewis lung carcinoma (LLC) on mice. The acute toxicity of the tested compounds was appointed in C57BL mice and the antitumor effect on LLC was assessed from the percent increase in median lifespan of the treated animals over the untreated (control) (T/C%). RESULTS: The lactam steroidal esters presented lower toxicity and increased antineoplastic activity in vitro and in vivo compared to their respective alkylating components. An A-lactam testosterone ester namely: 17beta-hydroxy- 3-aza-A-homo-4alpha-androsten-4-one-p-N,N-bis (2chloroethyl) amino phenoxy acetate (ALT-CAPOA) performed significantly higher anticancer activity in vitro and in vivo. This compound generated 37.5% 90-day disease free survivors (cures) against LLC. CONCLUSION: These results indicate a high antitumor potential of lactam steroid alkylating esters depended on the alkylating moiety as well as on the modified steroidal carrier. Preclinical research supports that ALT-CAPOA generates well-tolerated toxicity as well as superior antitumor activity against NSCLC. These significant results call for further clinical development. PMID- 17415827 TI - Superoxide dismutases and p53 protein levels in blood cells of breast cancer patients. AB - PURPOSE: Radiation treatment of breast cancer (BC) often results in post-therapy complications. The undesired sequelae could be avoided by the diagnostic screening of biomarkers for prediction of ionizing radiation (IR)-linked injury of healthy tissues. PATIENTS AND METHODS: The expression of antioxidative defence enzymes CuZn- and Mn-superoxide dismutase (CuZnSOD, MnSOD) and tumor suppressor protein p53 was measured in blood cells of 19 women with BC (age groups 30-45 and 46-60 years) and respective controls. The proteins were detected by specific immunostaining and quantified by laser-scanning densitometry. RESULTS: Constitutive expression of CuZnSOD was significantly elevated in the group of BC patients (up to 254 arbitrary units, AU/mL) relative to the control group (105 130 AU/mL). The constitutive expression of MnSOD was elevated (up to 94 AU/mL) in the group of BC patients relative to the controls (53-56 AU/mL). p53 was also constitutively more expressed in BC patients (35-42 AU/mL) than in controls (32 33 AU/mL). Both MnSOD and p53 were inducible by (60)Co gamma-ray IR (up to 170 AU/mL and 51 AU/mL, respectively) in the BC patient group. The levels of IR induced p53 correlated inversely with MnSOD levels. CONCLUSION: The constitutive expression of all 3 proteins could be a useful biomarker for the presence of BC, but only MnSOD overexpression may be the predictive biomarker for selection of BC patients that would be less susceptible to IR-linked complications. PMID- 17415828 TI - Serum HER2 and CA 15-3 in breast cancer patients. AB - PURPOSE: The HER2 oncogene encodes a transmembrane tyrosine kinase receptor. This molecule could be a new marker for prognosis and response to therapy in patients with advanced breast cancer. However, the extracellular domain of c-erbB-2 (HER2) transmembrane receptor undergoes proteolytic cleavage from the fulllength protein by metalloproteases, and is shed into the blood as a circulating antigen. To determine the clinical utility of this oncoprotein, the soluble form of HER2 was assayed in the serum of breast cancer patients. PATIENTS AND METHODS: Serum levels of breast carcinoma antigens CA 15-3 and HER2 were determined in 60 patients, 40 with localized (group A) and 20 with metastatic (group B) breast carcinoma. CA 15-3 measurements served as "gold standard" to which HER2 diagnostic and/or prognostic value was compared. Sera from 10 healthy women served as controls. RESULTS: Serum levels of the tested tumor markers HER2 and CA 15-3 were significantly higher in cancer patients compared to controls. CA 15-3 correlated with bulky initial tumor, whereas HER2 showed no differences between healthy individuals and group A patients. The serum levels of the tested markers in group B patients were significantly higher (p <0.001) than the serum levels of patients in group A. Striking increase in serum levels of HER2 was found in 52.7% and CA 15-3 in 52.9% of patients with metastatic cancer. A combination of markers was more sensitive than using one marker alone. In this regard, 90% of the patients with metastasis had at least one of the markers increased. CONCLUSION: The results of this study suggest that the HER2 oncoprotein may be potentially useful in detecting recurrence of breast cancer. However, to improve sensitivity and specificity in the diagnosis and monitoring of breast cancer, the use of multiple tumor markers should be employed. PMID- 17415829 TI - Hormone receptors in benign intracranial meningiomas. AB - PURPOSE: Although some embryologic data suggest that the neural crest elements, which are not a target tissue for estrogen and progesterone, participate in the forming of meningothelial cells, meningiomas show hormone receptors. The aim of this study was to investigate the estrogen (ER) and progesterone receptors (PR) content of benign meningiomas (WHO grade I) which were completely resected. MATERIALS AND METHODS: Paraffin-embedded tissue sections of 30 intracranial meningiomas were examined immunohistochemically using monoclonal antibodies for ER and PR. RESULTS: All 30 (100%) tumor samples were ER-negative. Seventeen (57%) of them were positive for PR. These receptors were found in 8 of 17 (47.50%) tumors in female patients and in 9 of 13 (69.23%) tumors in male patients (Mann Whitney p <0.05). Age, localization of tumors and histological subtype did not correlate with PR status. CONCLUSION: This study demonstrated complete absence of ER in benign meningiomas. In contrast, PR were positive in more than half of the patients. PR positivity was significantly higher in male patients. PMID- 17415830 TI - Diagnosis and follow up of primary ocular melanoma by radioimmunoscintigraphy. AB - PURPOSE: To determine the usefulness of immunoscintigraphy with Tc-99m labeled anti-melanoma antibodies in the diagnosis of primary ocular melanoma. Furthermore, to try to determine whether the treatment of ocular melanoma by contact radiotherapy had an influence on the immunoscintigraphic disease detection. PATIENTS AND METHODS: Immunoscintigraphy was performed on 17 patients: 9 patients with newly diagnosed choroidal melanoma (group A), 6 patients with choroidal melanoma treated by ruthenium contact radiotherapy (group B) and 2 patients with melanoma conjunctivae (group C). F(ab')2 fragments of 225.28S monoclonal antibody directed against a high molecular weight melanoma- associated antigen (HMW-MAA) were used. Planar scintigtaphy was done 4 hours after intravenous (i.v.) injection of 300-445 MBq of labeled antibodies, in anterior Waters projection, with 95 degrees of cranial tilt of the gamma-camera head. 500,000 counts static images were acquired in 256x256 resolution matrix. RESULTS: All group A patients showed positive immunoscintigraphy. The maximum diameter of these tumors, measured by ultrasound, was between 9 and 18 mm, with maximal prominence between 5 and 10.5 mm. Both patients of group C showed negative immunoscintigraphy, due to the small dimensions of the tumors (less than 2.7 mm). In all 6 patients of group B immunoscintigraphy was negative although the dimensions of the tumors were sufficient enough to be visualized by immunoscintigraphy (diameter between 8.3 and 17 mm and prominence between 5 and 10.3 mm). CONCLUSION: According to our preliminary results, we concluded that immunoscintigraphy was useful diagnostic modality in the detection of small intraocular tumors. The negative immunoscintigraphy in patients with tumors treated by contact radiotherapy could be explained by the biological regression of the tumor that precedes regression of its volume. This finding indicates that immunoscintigraphy could be useful in the follow-up of conservatively or surgically treated patients with choroidal melanoma. PMID- 17415832 TI - Lymphoma of the breast - case report and review of the literature. PMID- 17415831 TI - Beam characteristics of kilovoltage radiotherapy unit. AB - PURPOSE: To present the beam characteristics of the Siemens II orthovoltage x-ray therapy machine. MATERIALS AND METHODS: x-ray beam qualities with the tube operating between 100 and 300 kV were determined. The field flatness, central axis depth dose, and relative output factors were measured for several applicator sizes for rectangular cones at 40 cm, and for circular cones at 30 cm focal skin distance (FSD). RESULTS: The central axis depth doses were selectively compared with values presented in the literature and found to agree to within -/+3 % shallow depths for two energy ranges. The field edges had 85-95 % isodose line. Relative output factors of all the available cones were different. CONCLUSION: The beam quality for all the stations should be determined and relative output factors should be measured for all the available cones. PMID- 17415833 TI - Successful palliative treatment of an inoperable giant squamous skin cancer: a case report. AB - A 63-year-old male patient with locally advanced stage giant squamous cell skin cancer in the back was admitted to our clinic. He was treated by radiotherapy with palliative intent and achieved a nearly complete response. However, the tumor recurred 3 times after various palliative treatments (surgery, chemotherapy). The tumor finally progressed and the patient died 28 months after his first admission to the clinic. We present this case because of the unexpectedly excellent response to radiotherapy and achievement of successful palliation with other treatment modalities. PMID- 17415834 TI - A case of melanoma metastatic to the gallbladder and the common bile duct with clinical presentation of obstructive jaundice and bilirubinemia. AB - Cutaneous melanoma is known to have the capacity to metastasize to virtually any organ. Because melanoma metastases to the gallbladder and the extrahepatic bile ducts are usually clinically asymptomatic, there are only few reports on this topic in living patients. We describe a case of melanoma metastatic to the gallbladder and the common bile duct presenting with obstructive jaundice and bilirubinemia. PMID- 17415835 TI - Human immunodeficiency virus-negative tonsil Kaposi's sarcoma and Hodgkin's disease: case report and review of the literature. AB - Kaposi's sarcoma (KS) is a multicentric malignant neoplastic vascular disorder characterized by multiple violet-colored nodules of the skin. The coexistence of KS with other primary malignancies, especially of the lymphoreticular system, has been frequently noted. However, the association of Hodgkin's disease with KS is a rare occurrence. In this article we present the case of a 33-year-old man with human immunodeficiency virus (HIV)-negative KS of the tonsil, occurring in the radiotherapy field for Hodgkin's disease treated 20 years ago. PMID- 17415837 TI - Rudolf virchow (1821-1902): founder of cellular pathology and pioneer of oncology. AB - Rudolf Virchow, distinguished pathologist, physical anthropologist, and statesman, was probably the most distinguished German pathologist of his age, and is regarded as the founder of Cellular Pathology. He contributed greatly to the study of tumors, leukemia, hygiene, and sanitation. PMID- 17415836 TI - A case of exocrine-endocrine neoplasm of the pancreas in a patient with ulcerative colitis with literature review. AB - Until now only Japanese authors have reported 4 cases of pancreatic neoplasm associated with ulcerative colitis (UC). We report on a case of a 44-year-old woman who was operated on for complicated UC and an exocrine-endocrine neoplasm of the pancreas, where the endocrine component was presented by pancreatic polypeptide (PP)-producing cells. By means of molecular genetics methods we found microsatellite instability (MSI) in the markers D18S35, FGA and p53 in the colonic lining, and loss of heterozygosity (LOH) in the p53 marker in the pancreatic tumor. A literature review concerning the coexistence of these two conditions showed that PP is involved in the pathogenesis of the UC and that UC increases the risk of development of extracolonic neoplasms. PMID- 17415838 TI - The past and the future of cancer genetics. AB - Carcinogenesis represents a multistep process associated with accumulation of somatic mutations in the classes of genes that regulate cell proliferation, apoptosis as well as DNA repair. Oncogenes, positive regulators of cell proliferation are activated during carcinogenesis. On the contrary, tumor suppressor genes, negative regulators of cell proliferation have to be inactivated. Mutations in genes that function in the maintenance of genomic stability are manifested by increase in the mutation rate in cancer cells that drive tumor progression. In general, on the basis of malignant transformation lies the abrogation of the balance between cell proliferation and cell apoptosis. The genetic mechanisms included in the transformation of normally acting genes comprise a wide spectrum of events, such as gene mutation, gene and chromosome rearrangement and gene amplification. Besides the role of somatic gene alteration in the development of sporadic cancer, germline mutations are the basis of a substantial number of inherited cancer syndromes. The future decades will be marked with the expansion of data exploiting cancer genetics, epigenetics and genomics into clinical practice. Consequently, translational cancer research should provide the generating of new targeted therapies, since individual molecular profiling of a patient?s tumor should increase efficacy of conventional anticancer therapies such as chemotherapy and radiotherapy. PMID- 17415839 TI - Is Open Access to medical research literature relevant to low- and middle-income countries? AB - At the end of 2003, the United Nations held the first phase of the World Summit on the Information Society, the aim of which is to develop a better understanding of the international impact of the information and communication technology revolution. Part of the first meeting's declaration referred to the Open Access movement. This article describes what Open Access is, its implications for medical research in low- and middle-income (LAMI) countries, and considers whether Open Access will make a difference. PMID- 17415840 TI - Good Scientific Practice. Part IV. Conflict of interest. PMID- 17415841 TI - The status of chemoprevention in the current cancer therapeutic armamentarium. AB - Cancer mortality nowadays remains unacceptably high despite immense advances in the understanding of the mechanisms of carcinogenesis, in bringing potent new drugs to the clinic and in treating several rare forms of cancer. Many scientists suggest that overall cancer mortality statistics are unlikely to change in a fundamental way until there has been a re-orientation of emphasis in cancer research that will direct greater resources towards prevention of disease development, rather that treatment of end-stage disease. Cancer chemoprevention represents a rather new rational approach in the management of cancer. Although the results of chemoprevention clinical trials will appear in the near future, the current preclinical and initial clinical published data outline the significant future perspective of cancer chemoprevention. PMID- 17415842 TI - Carcinoma in situ of the urinary bladder. AB - Urinary bladder cancer ranks 4th among cancers in men and 8th in women. Superficial bladder cancer (stages Tis, Ta and T1) accounts for 75% of bladder cancer cases, whereas carcinoma in situ (CIS) may appear as an entity of its own, or coexist with another tumor. The diagnosis of CIS is based on urine cytology, cystoscopy and biopsy. Long term follow-up is mandatory due to its high recurrence rate and its invasive potential. Because of this, radical cystectomy has been the gold standard treatment till the mid 1980's. The emergence of intravesical infusion of drugs, especially of BCG, changed the management of CIS altogether. Today, BCG infusion after the initial transurethral resection (TUR) is considered the best treatment, although a consensus regarding the ideal dosing scheme or the maintenance scheduling, has not yet been reached. This, as well as newly developedtherapeutic means, underscore the need for further study on the ideal treatment of CIS of the urinary bladder. PMID- 17415843 TI - The RACOX phase I study: radiation (RA), capecitabine (C) and oxaliplatin (OX) as adjuvant treatment of stage II and III rectal cancer. AB - PURPOSE: The aim of this phase I trial was to deter- mine the maximum tolerated dose (MTD) of adjuvant che- motherapy (CT) with oxaliplatin in combination with capecitabine during concomitant pelvic radiotherapy (RT) in patients with rectal cancer. PATIENTS AND METHODS: Eligible patients had pathological stage II (T3 4N0M0) or III (any T N1-2M0) rectal adenocarcinoma, and no prior treatment other than curative resection. Fixed capecitabine dose (825 mg/m(2) bid on days 1-14 and 22-35) was given and external beam RT was delivered to the pelvis (50.4 Gy in 27 fractions in 5.5 weeks, with field reduction after 45 Gy in linear accelerator, 18Mev). Oxaliplatin was tested at 4 dose levels: 100, 110, 120 and 130 mg/m(2). The dose of oxaliplatin was escalated when all 3 entered patients at each level had been monitored for at least 8 weeks after the CT/RT course without dose limiting toxicities (DLTs). In the presence of a DLT at any dose level, a further 3 patients were enrolled. If only 1 of the 6 patients experienced a DLT, escalation could proceed. The MTD was defined as the level at which >/= 2 of 3 to 6 patients experienced DLTs. Fifteen patients (10 males and 5 females, median age 62 years) were enrolled at oxaliplatin dose levels of 100 (n=3), 110 (n=3), 120 (n=3) and 130 mg/m(2) (n=6). RESULTS: All patients completed the planned CT/RT course. Dose reduction or delay of the 2nd CT cycle was not required. No DLTs were observed at all dose levels. Overall, gastrointestinal and neurological toxicities were mild and transient. Toxicities included non-dose-limiting nausea / vomiting, diarrhea, dysesthesias in 2 level III and in 1 level IV patients. Grade II myelotoxicity, mainly neutropenia, was seen in 6 patients. With a median follow-up of 4 months (range 2-12) after the completion of CT/RT, late toxicities were restricted to grade II radiation colitis and dermatitis in 2 and 2 patients, respectively. CONCLUSION: The combination of pelvic RT, capecitabine and 3-weekly oxaliplatin is feasible and well tolerated. The MTD was not reached up to the dose of 130 mg/m(2) of oxaliplatin, which is the recommended dose. PMID- 17415844 TI - Intraoperative electron beam radiotherapy followed by moderate doses of external beam radiotherapy in the treatment of resected soft tissue sarcomas of the extremities. AB - PURPOSE: Soft tissue sarcomas (STS) have a high incidence of local recurrence. In an effort to improve the local control rate and the survival in patients with STS, treatment strategies employing intraoperative electron beam radiotherapy (IOERT) in combination with external beam radiotherapy (EBRT) and extensive surgical resection have been explored. This study assesses the rate of overall survival (OS), local control and toxicity of this multimodal approach for primary and recurrent STS of the extremities. PATIENTS AND METHODS: From 1999 to 2004, 36 patients were treated at Agios Savvas Cancer Hospital for primary or recurrent extremity STS with IOERT as a component of their treatment. All patients underwent surgical resection, IOERT, and most of them received postoperative EBRT with a median dose of 45 Gy. Chemotherapy was given to patients with high grade tumors. Thirteen patients were treated for primary disease and 23 for isolated local recurrence.The locations of the tumors were as follows: upper limbs n=19, lower limbs n=17. Tumor size was >5 cm in 16 (44%) patients and high-grade histology (II-III) was present in 24 (67%) patients. Six (17%) patients had positive surgical margins. RESULTS: With a median follow up of 24 months (range 6 48) OS was 72% (84.5% for patients with low grade lesions compared to 65% for high grade lesions, p=0.127, and 90% for tumors <5 cm compared to 50% for tumors >5 cm, p=0.0136). Overall local tumor control rate was 89% (92% in primary disease group versus 87% in isolated local recurrence group, p=0.136, and 93% for patients with negative surgical margins versus 67% for those with positive margins, p=0.0013). Distant metastases occurred in 10 patients (1 of 13 (8%) with primary disease, and 9 of 23 (39%) with isolated local recurrence). All distant metastases were to the lungs. Twelve (33%) patients developed moderate neurotoxicity. CONCLUSION: In selected patients, IOERT results in excellent local control and OS with acceptable toxicity. PMID- 17415845 TI - Local administration of interferon-alpha in cases of cervical intraepithelial neoplasia associated with human papillomavirus infection. AB - PURPOSE: To investigate the efficacy and the optimal dose of topical interferon alpha in cases of cervical intraepithelial neoplasia (CIN) associated with human papillomavirus (HPV) infection. PATIENTS AND METHODS: From January 1995 to December 1997, 75 patients with CIN (45 with CIN I-III, and 30 with CIS) associated with HPV infection were treated with local administration of interferon-alpha (group 1). From January 2002 to June 2003, after polymerase chain reaction (PCR) determination of HPV types 6, 11, 16, 18, 31 and 33, treatment with interferon-alpha was administered to 21 patients with CIN I-II (group 2). HPV 16 was found in 11 cases and HPV 31 in 10 cases. In cases of CIS, the interferon treatment followed conization. In all cases interferon was injected intralesionally or at periphery of the conization, 3 times per week at dose of 3 million (m) IU. Ten interferon administrations were given in all of the cases. RESULTS: In group 1 after 5 administrations the cytological findings returned to normal in 48 (64%) cases, and in 27 (36%) cases only dyskaryotic cells were found. In all cases the cytological findings reverted to normal after 10 administrations of interferon. In 45 cases with CIN I-III treated with interferon only (without conization), biopsy was carried out, and all of the patients were found free of dysplastic lesions. In group 2 the cytological findings of all 21 cases returned to normal after 10 administrations of interferon. As of December 2003, both the cytological and histological findings were negative for CIN and HPV. CONCLUSION: These results suggest that treatment with interferon-alpha is an effective therapeutic method for CIN associated with HPV infection, and 10 administrations of 3 mIU constitute the optimal dose. PMID- 17415846 TI - Localization, clinical and pathological characteristics and survival in sporadic colon cancer patients younger than 40 and over 65 years of age. AB - PURPOSE: Colorectal cancer is predominantly a disease of older population, but occasionally it affects younger patients, in whom very often diagnosis is overseen and treatment begins late. The aim of our study was to compare localization, clinical and pathological characteristics and survival of sporadic colorectal cancer patients aged up to 40 and over 65 years. PATIENTS AND METHODS: The first group (group I) included 19 patients under 40 years and the second group (group II) 28 patients aged over 65 years, treated during 1997-2001. Patients with family history of colon cancer and inflammatory disease of the colon were not included. Arithmetic mean, standard deviation, Fisher's test, Student 's t test, x(2) test and the Kaplan-Meier method were used in the statistical analysis of the results. RESULTS: There was no difference among the tested groups regarding tumor localization. The most frequent localization was in the rectum and left colon. At presentation, in group I patients, besides the metastases in the liver and lymph nodes, colorectal cancer infiltrated also the duodenum, stomach, right kidney capsule in one patient, and adnexa in two patients. In group II patients we registered only liver and lymph node metastases. Pathologically, tubular and mucinous forms were present in all of the patients up to 40 years of age, while only one patient over 65 had tumor with mucinous component. In group I, Astler-Coller stage B was found in 1.5% of the patients, stage C in 72.5% and stage D in 26%; in group II, stage B was found in 1.5%, stage C in 84.5% and stage D in 14%. Grade III was 36.8% in group I and 17.8% in group II. No statistical differences were found in stage distribution (p=0.36) and grade (p=0.06) between group I and II. Five-year overall survival was 57.8% and 28.5% in younger and older patients, respectively (p=0.053). CONCLUSION: The results obtained showed no difference in clinical symptomatology and tumor localization in both groups. The incidence of more aggressive tumors was higher in younger persons. However, early detection combined with more aggressive therapeutic approach, could enable significant improvement of the 5 year survival of younger patients with colon cancer. PMID- 17415847 TI - Evaluation of pulmonary complications of radiotherapy in breast cancer patients. AB - PURPOSE: To study the early and late pulmonary complications of radiotherapy (RT) in patients with operated breast cancer who received postoperative RT. PATIENTS AND METHODS: Radiation pneumonia (RP) and radiation fibrosis (RF) rates were evaluated after 3 and 18 months from the end of RT, using the Radiation Therapy Oncology Group and the European Organization for the Research and Treatment of Cancer (RTOG/ EORTC) combined toxicity classification scale. Evaluation included physical examination, high resolution computed tomography (HRCT) of the thorax, and pulmonary function tests (PFTs). The incidence of RP and RF, the relationship between RP and RF and possible predisposing factors and the impact of RT on the PFTs were analyzed. RESULTS: Between December 2000 and March 2001 35 patients were included in the study. Due to several reasons 29 patients were evaluable for RP and 25 for RF. On the 3rd month post-RT, 17 (59%) patients developed RP. Three (18%) of the cases were grade 1, 13 (76%) grade 2 and 1 (6%) grade 3. One patient was radiologically diagnosed with early RF. When evaluated for RF, 9 (36%) of 25 patients were found to have RF. Four (45%) of them were grade 1, and 5 (55%) grade 2. FEV1, FEV1/FVC, VC, FVC values showed different degrees of decline on the 3rd month. The most prominent change occurred with DLCO/ VA ratio which decreased by 20%. On the 18th month, all values returned to at least the pretreatment levels. CONCLUSION: RT-induced RP and RF are quite frequent. As clinical findings are generally nonspesific, radiological findings of RP and RF should be known. Early and late effects on PFTs are generally mild and transient. PMID- 17415848 TI - Survival of patients with resected pIIIa - N2 non small cell lung cancer: suggestion for subclassification. AB - PURPOSE: The group of completely resected stage IIIa- N2 non small cell lung cancer (NSCLC) patients is considered heterogeneous in various aspects including survival and pattern of recurrence. The prognostic factors still remain controversial. Clinical trials dealing with multimodal strategy for N2 NSCLC are being watched with keen interest, and the feasibility of this strategy is to be confirmed. In the present study we attempted to clarify the role of different clinicopathological factors which separate patients into high and low- risk groups based on the disease-free and overall survival. PATIENTS AND METHODS: The study comprised 60 consecutive patients with pathologically (p) proven N2 NSCLC who had undergone complete surgical disease resection with curative intent between January 1997 and September 2000. All patients had an apparently resectable disease at preoperative staging and thoracotomy. Extensive mediastinal lymph node dissection was performed when possible and consisted of removal of all ipsilateral mediastinal lymph nodes. Patients were submitted to postoperative split course adjuvant radiotherapy. Cumulative survival rates were calculated by the Kaplan-Meier method. The analyzed prog- nostic factors were age, histological type and grade of differentiation, clinical (c) stage, cN status and tumor size and were tested for statistical significance by univariate analysis using log rank test and Willcoxon test. RESULTS: The median follow-up period was 36 months (range 24-72 months). Tumor size, cN status and age were significant predictors of survival. Large tumor size (T3) was significantly worse predictor compared to T1 (p=0.046) and cN2 status evaluated by computed tomography (CT= also showed statistically significant unfavorable prognosis in comparison with cN0 and cN1 (p=0.0185). Patients over 65 years had significantly worse prognosis compared to those under 65 years (p=0.008). CONCLUSION: This study identified stage IIIa - N2 NSCLC prognostic subgroups and suggests different therapeutic approach according to the subgroup profile. PMID- 17415849 TI - Diagnostic procedures of pleural malignant mesothelioma: our experience. AB - PURPOSE: The morphology of the epithelioid malignant mesothelioma (MM) of the pleura is similar to lung adenocarcinoma involving pleura. The aim of this study was to evaluate the value of immunohistochemistry in the accurate diagnosis of MM, especially of the epithelioid type with needle biopsy of the pleura. MATERIALS AND METHODS: The diagnosis of MM was established with pleural needle biopsy and tumor immunophenotyping in 30 patients. A broad spectrum of monoclonal antibodies was applied: HBME-1, E-cadherin, calretinin, cytokeratin 5/6, vimentin, thyroid transcription factor (TTF-1) and surfactant apoprotein A (SP A). RESULTS: We diagnosed 24 epithelioid, 2 biphasic and 4 sarcomatoid MM. HMBE-1 was the most sensitive tumor marker of the epithelioid type, being positive in 100% of the cases. Calretinin, E-cadherin and cytokeratin 5/6 were positive in 70%, 73%, and 50% of all tumors, respectively. TTF-1 and SP-A were negative in all MM. Vimentin was positive in spindle cells of all sarcomatoid and biphasic MM (20%). CONCLUSION: The accurate diagnosis of MM is mandatory for appropriate treatment decision (surgical or nonsurgical). Our results demonstrate that HMBE-1 is a most useful diagnostic antibody for epithelioid MM, and TTF-1 for lung adenocarcinoma (its thyroid origin excluded) involving pleura. PMID- 17415850 TI - Expression of galectin-3 and carcinoembryonic antigen in the trophoblast of the gestational trophoblastic disease. AB - PURPOSE: Galectin-3, an endogenous beta-galactoside- binding protein, has been implicated in the regulation of many physiological and pathological cellular processes through specific interactions with complementary ligands. The level of galectin-3 expression has been correlated with metastatic potential in many tumor types. The present study was designed to investigate possible correlation of the expression of galectin-3 with carcinoembryonic antigen (CEA), one of the putative galectin-3 ligands, in the transformed trophoblast of the gestational trophoblastic diseases (GTDs) compared to the invasive trophoblast (interstitial and endovascular) of the normal first trimester of pregnancy placenta. MATERIALS AND METHODS: Double immunohistochemistry was used to examine paraffin sections of 9 specimens of normal placenta, 8 cases of invasive mole and 7 cases of choriocarcinoma. Immunoreactivity was scored semiquantitatively to include both the percent of trophoblast cells stained as well as the intensity of staining. RESULTS: Data presented here demonstrate similar patterns of expression for galectin-3 and CEA in transformed GTDs trophoblastic cells, as well as in normal invasive trophoblast. Upregulation of galectin-3 in transformed GTDs trophoblast was followed by high levels of CEA expression in the same tissue specimen (significant differences at < 0.05, Mann-Whitney Rank Sum Test). CONCLUSION: These results may point to the CEA as one of the ligands for galectin-3 in transformed trophoblast, which merits further investigation. PMID- 17415851 TI - Cytotoxic effects of a mixed ligand copper(II) chelate complex against a panel of human and murine cancer cells in vitro. Theoretical study of the mechanism of biologic action through molecular modelling. AB - PURPOSE: In an effort to discover new compounds with anticancer activity, we have developed a novel copper (II) [Cu(II)] chelate complex with a tridentate ONNSchiff base ligand and the anion of salicylate and we evaluated the in vitro chemosensitivity of various human and murine tumor cell lines by measuring cell growth inhibition. The ultimate goal was to evaluate the existence of a potential antitumor activity of this complex. Beyond the cytotoxic activity assessment of the complex, we aimed at the elucidation of the underlying mechanism of action of this complex and its interactions with biological molecules, carrying out theoretical (quantum-chemical) calculations. MATERIALS AND METHODS: Cells grown in adherence or in suspension in 96-well microplates were exposed to Cu(II) complex for 24, 48 or 72 h. In vitro drug cytotoxicity was assessed by SRB and XTT colorimetric assays. Molecular modelling tools were used applying semiempirical and ab initio calculations. RESULTS: A series of experiments was carried out, showing a potent cytotoxic activity against most of the tested cancer cell lines. Quantum-chemical calculations demonstrate that the mechanism of the cellular damage can be explained, at least in part, by the ability of the nucleobases and nucleotides to be subject to nucleophilic attack on copper. CONCLUSION: Profound growth inhibitory effects were observed for the tested Cu(II) complex. It was also verified the hypothesis that the mechanism of action of this complex involves binding to DNA and RNA. These findings prompt to search for possible interaction of this complex with other cellular elements of fundamental importance for cell proliferation. PMID- 17415852 TI - Antioxidative and antiproliferative effects of Satureja montana L. extracts. AB - PURPOSE: To study in vitro the antioxidative effect of 6 Satureja montana L. extracts on free radicals and their antiproliferative effect on human tumor cell lines. MATERIALS AND METHODS: The antioxidative effect of extracts on 2, 2 diphenyl-1-picryhydrazyl (DPPH) radical was investigated by electron spin resonance (ESR) spectroscopy. Cell growth effect was measured by sulforhodamine B colorimetric assay on HeLa (human cervix epidermoid carcinoma), HT-29 (human colon adenocarcinoma), and MCF-7 (human breast adenocarcinoma) cell lines. IC(50) values were calculated from the concentration response curves following 48 h exposure time. RESULTS: The antioxidative activity of extracts increased dose dependently at mass concentrations ranging from 0.05 to 0.3 mg/ml, and decreased in the following order: n-butanol > methanol > water > ethyl acetate > petroleum ether. All extracts effected cell growth but in a different way, depending on the extract dose and cell line. Extracts exhibited antiproliferative effect on HeLa cell line with IC(50) values ranging from 0.41 to 0.84 mg/ml except petroleum ether (IC(50) >1 mg/ml). Petroleum ether and chloroform extracts stimulated proliferation of HeLa cells within a concentration range from 0.0625 to 0.125 mg/ml. No extract reduced MCF-7 cells growth by 50% even at the concentration of 1 mg/ml. Only petroleum ether and chloroform extracts induced significant growth inhibition of HT-29 cells (IC(50) was approximately 0.74 mg/ml for both extracts). Strong stimulation of HT-29 proliferation was observed within a concentration range from 0.0625 to 0.25 mg/ml for petroleum ether, n-butanol and chloroform extract, and from 0.0625 to 0.5 mg/ml for methanol and water extracts, respectively. CONCLUSION: The obtained results indicated that Satureja montana L. extracts are strong antioxidants in vitro. ESR data demonstrated that n-butanol, methanol and water Satureja montana L. extracts possess high antioxidative activity. Chloroform extract did not show any antioxidative activity. Satureja montana L. extracts selectively inhibited the growth of human tumor cells. PMID- 17415853 TI - A comparison of Monte Carlo simulation with experimental dosimetric techniques for a 6 MV stereotactic radiotherapy unit. AB - PURPOSE: To compare Monte Carlo simulation with conventional dosimetry techniques for stereotactic radiotherapy (SRT), since accurate dosimetry of narrow photon beams is very complicated and has often been questioned, mainly due to the lack of lateral electronic equilibrium and uncertainty in beam energy in terms of steep dose gradients. MATERIALS AND METHODS: In this work a Monte Carlo (MC, EGS4) simulation for dosimerty study was performed for the 6MV home made SRT unit of the University Hospital of Patras (Hellas). The results were compared with conventional small field dosimetry techniques such as ionization chamber, TLD's, and films (conventional and radiochromic). Hence, a comparison of many of the dosimetric techniques currently being used in small field dosimetry was attempted. RESULTS: It was shown that all techniques are in reasonable agreement (within -/+ 2%) and that Monte Carlo can be used as a reliable reference for the dosimetry of the SRT beams, especially where lateral electronic equilibrium does not exist, as long as accurate simulation can be achieved. CONCLUSION: This study is only limited by the insurance of accurate simulation of the linear accelerator, which can be a difficult task since it is limited by the availability of the manufacturer's designs and the availability of computers and computer time for adequate runs, but it could become a useful tool for Monte Carlo simulations, as it contains detailed analysis of the run parameters and component modules selection. PMID- 17415854 TI - Paget's "seed and soil" hypothesis revisited. AB - Paget's "seed and soil" hypothesis emphasises the importance of the interaction between the tumour cell and its environment in order for metastasis to occur. An unusual case of metastatic renal carcinoma is described with review of the literature surrounding the "seed and soil" hypothesis. PMID- 17415855 TI - Peutz-Jeghers syndrome combined with breast cancer, cervical carcinoma and ovarian gonadoblastoma: a case report. AB - Peutz-Jeghers syndrome (PJS) is a rare autosomal dominant condition characterized by gastrointestinal polyps, mucocutaneous pigmentation and an increased risk for cancer. In this report, a 34-year-old woman with PJS associated with a rare ovarian tumor (gonadoblastoma) and synchronous breast and cervical carcinoma is discussed and the relevant literature is reviewed. PMID- 17415856 TI - Ovarian fibrothecoma associated with Meigs' syndrome and elevated serum CA 125. AB - Meigs' syndrome consists of a benign ovarian tumor accompanied by ascites and hydrothorax. Elevated serum CA 125 in postmenopausal women with a solid adnexal mass, ascites and pleural effusion is highly suggestive of a malignant ovarian tumor. We report on an unusual case of a benign fibrothecoma of the ovary associated with Meigs' syndrome and elevated CA 125 level in an 62-year-old white female. Although rarely, a benign ovarian tumor should be considered in the differential diagnosis of an adnexal mass associated with Meigs' syndrome and elevated serum CA 125 levels. PMID- 17415857 TI - Atypical presentation of an unusual lung tumor. AB - We describe a case of a rare bronchial epithelial- myoepithelial carcinoma of the lung in a 36-year-old man. Tumor enucleation was carried out and the patient was put on long-term follow up. Tumor categorisation was difficult. The patient is disease-free on the 30th postoper- ative month. The clinicopathological uniqueness of this neoplasm is reviewed. PMID- 17415859 TI - Paul Ehrlich (1854-1915): founder of chemotherapy and pioneer of haematology, immunology and oncology. AB - The Jewish-German histologist, microbiologist, bio- chemist, immunologist and pharmacologist Paul Ehrlich, by his research founded chemotherapy and pioneered im- munology, haematology and oncology. For his contribu- tion in immunology he was awarded the Nobel Prize of Medicine and Physiology in 1908. PMID- 17415858 TI - Primary cerebral gliosarcoma: a case presentation with review of the literature. AB - In this presentation we describe a rare case of a 42- year-old female with a large right frontal gliosarcoma (GS) treated with gross total resection of the tumor and postoperative external radiotherapy. The patient did not respond to treatment and she died 3 months after the end of radio- therapy. We also present a review of the literature on epidemiology, pathogenesis, clinical presentation, diagnosis and treatment of this uncommon clinical entity. PMID- 17415860 TI - Good Scientific Practice: ethical codex of science. I. Formulating ethical codex of science: A Pan-European initiative. PMID- 17415861 TI - The role of endocrine therapy in early breast cancer: adjuvant treatment of preand postmenopausal patients. AB - Two major trends are evident from ongoing conferences and panels of experts on early breast cancer adjuvant treatment. The first strongly suggests the application of wellestablished standards of adjuvant treatment in everyday clinical practice. The second encourages participation in international adjuvant clinical studies of breast cancer. Our growing knowledge of the role of endocrine adjuvant therapy in early breast cancer has emerged from the findings of large adjuvant studies. Meta-analyses of numerous randomised trials have accumulated evidence that has been analysed on many occasions by international panels of experts and has become a major source of consensus recommendations. Nevertheless, many open questions still exist, and new ones arise. This paper reviews the current standards for the adjuvant treatment of endocrine-dependent breast cancer, as well as some perspectives emerging from ongoing clinical studies. PMID- 17415862 TI - Paclitaxel or docetaxel combined with platinum in advanced ovarian cancer? AB - Epithelial carcinoma of the ovary is one of the most common gynecologic malignancies and ranks 5th as a cause of cancer death in women, with half of all cases occurring in women aged over 65. Because ovarian cancer is often asymptomatic in its early stages, most patients have widespread disease at the time of diagnosis. Surgery is required for correct diagnosis and accurate staging, representing also the main form of treatment for early-stage disease which is confined to the ovaries. In advanced stages (III and IV) where the tumor has spread beyond the pelvis, the initial surgical treatment is followed by chemotherapy. Ovarian tumors are sensitive to chemotherapy, and most stage III and IV patients receive chemotherapy to increase survival, disease-free interval and improve quality of life (QoL). Until the mid 1990s standard chemotherapy of advanced ovarian cancer involved a platinum compound, either cisplatin or carboplatin, administered either alone or in combination with cyclophosphamide. New therapy standards emerged after the publication of a trial by the Gynecologic Oncology Group ( GOG) in 1996 and the first reports of an intergroup trial in 1998 that confirmed the GOG results. These studies demonstrated that patients treated with cisplatin/paclitaxel had significantly higher response rates, progression-free survival and overall survival compared with the previous standard treatment of cisplatin plus cyclophosphamide. PMID- 17415863 TI - Is there an alternative approach to abdominoperineal resection for the treatment of very low lying rectal cancer? Five-year results. AB - PURPOSE: To investigate whether sphincter-saving surgery combined with preoperative radiotherapy and postoperative adjuvant chemotherapy can compare successfully with abdominoperineal resection in the treatment of very low lying rectal cancer. PATIENTS AND METHODS: Two groups of patients were studied. Group I included 65 patients (T2-3 N0-2 M0) with very low lying rectal cancer treated since 1995 by a sphincter- saving surgical procedure. All of them received preoperatively external radiotherapy and postoperatively adjuvant chemotherapy with 5-fluorouracil (5-FU) and folinic acid. This group was prospectively evaluated. Group II included 122 patients with the same tumor stage and location as Group I, who were treated by abdominoperineal resection and were retrospectively evaluated. The rate of local recurrence was compared between the two groups. RESULTS: In Group I patients the local recurrence rate was 13.84% (9 patients) for a period ranging from 6 months to the end of the second year after the operation. This group of patients were followed up to 60 months after the initial treatment. The local recurrence rate in Group II patients was 17.21% (21 patients; p >0.05). CONCLUSION: We consider that sphincter-preserving surgery combined with preoperative radiotherapy and postoperative adjuvant chemotherapy represents a successful alternative to abdominoperineal resection of very low lying rectal adenocarcinoma. PMID- 17415864 TI - Diagnostic possibilities of the electroexcision and cold knife conization for precancerous lesions of the uterine cervix. AB - PURPOSE: The aim of the present study was to assess and compare the diagnostic possibilities of the loop electroexcision procedure (LEEP) and of conventional surgical conization of the uterine cervix in women with cervical precancerous lesions. PATIENTS AND METHODS: The study involved 80 women aged between 22 and 63 years (mean age 38.75 years), with cytological findings indicating various degrees of dyscaryosis. The patients were subjected to colposcopic targeted biopsy with subsequent (within 3 months) wide excision (40 patients with LEEP and 40 with cold knife conization). RESULTS: In both groups of women coincidence of the results to 1 degree of difference was found in 34 (85%) women. Analysis of the results obtained showed no statistically significant difference (p >0.05), both in the cases of full agreement as well as in the cases with 1 degree of difference, between the findings obtained by LEEP and surgical conizations on the one hand, and targeted biopsies on the other hand, as well as between the two excisional procedures. Also, no statistically significant difference between LEEP and surgical conization was found in comparing the frequencies of resection lines in healthy tissues and in the cases with visible squamous/ columnar junction (p= 0.418). At the same time, such a difference was found in comparing the cases with invisible junction (p= 0.0003), which was due to the cases with positive edges after LEEP. CONCLUSION: The results of the present study have evidenced the good possibilities of the LEEP and surgical conization as precision diagnostic tools in precancerous lesions of the uterine cervix. Based on the results obtained we think that LEEP has advantages in lower-grade lesions with visible boundary between squamous and columnar epithelium. In all other cases the method of choice should be surgical conization. PMID- 17415865 TI - T cell receptor-mediated signalling in T cells from multiple myeloma patients. AB - PURPOSE: Several abnormalities of the T-cell receptor (TCR) CD3 transduction pathway in cancer patients including B-cell neoplasms have been reported. Adequate activation of the receptor is important to provide stimulus for the T cell to start its effector functions. Activation of TCR/ CD3 results in recruitment of tyrosine kinases and changes of tyrosine phosphorylation levels. We investigated phosphorylation levels in T-cells from multiple myeloma patients at diagnosis and compared them to normal individuals. PATIENTS AND METHODS: We quantitatively analyzed protein phosphorylation of resting and anti-CD3 stimulated Tcells in 10 previously untreated multiple myeloma patients and 6 healthy donors using two-colour flow cytometry. The results were shown as a median fluorescence intensity values. A specific monoclonal antibody directed against phosphotyrosine was used. RESULTS: No significant differences between helper Tcells and cytotoxic T-cells and no significant differences between multiple myeloma patients and controls were found. CONCLUSION: The flow cytometry technique used in our experiment allows a quick insight into phosphorylation pattern following activation of T-cells. Further experiments combining this method with measuring the activity of particular tyrosine kinases of the activation cascade may bring the necessary information explaining the nature of T cell dysfunction. PMID- 17415866 TI - Serum estimation of carcinoembryonic antigen, beta(2)-microglobulin and calcitonin as tumor markers in primary lung cancer. AB - PURPOSE: The aim of this study was to estimate the diagnostic significance of carcinoembryonic antigen (CEA), beta(2)-microglobulin (beta(2)-MG), and calcitonin (CAL) as tumor markers in patients with lung cancer. PATIENTS AND METHODS: Sixty patients were included and CEA, beta(2)-MG, and CAL serum levels were measured before treatment, and one and 6 months after treatment. RESULTS: The results taken showed that CEA mean baseline level was 23.4 ng/ml (normal range 0-5 ng/ml and up to 8 ng/ml in smokers), elevated in 80% of small-cell lung cancer (SCLC) cases, in 93% of adenocarcinomas, in 84% of squamous-cell carcinomas, and in 84% of large-cell carcinomas. The mean beta(2)-MG level was 3.6 mg/l (normal range 0.7-3.4 mg/l), elevated in 70% of SCLC, in 40% of adenocarcinomas, in 66% of large-cell and in 50% of squamouscell carcinomas. The mean CAL level was 170 pg/ml (normal < 26 pg/ml), elevated in 60% of SCLC, in 33% of adenocarcinomas, in 16% of large-cell and in 16% of squamouscell carcinomas. CONCLUSION: CEA, beta(2)-MG, and CAL can be used as tumor markers with diagnostic value in primary lung cancer. PMID- 17415867 TI - Influence of venezuelan equine encephalitis, influenza and mumps vaccine viral strains on morphological patterns and viability of human tumor cells in vitro. AB - PURPOSE: To study the cytolytic and cytostatic effects in some human tumor cells lines after their infection with vaccine viral stains. MATERIALS AND METHODS: Cultures of human tumor cell lines (Hep-2, P3HR-1 and K-562) were exposed to vaccine viral strains of mumps, influenza and venezuelan equine encephalitis (VEE) to determine the impact of these strains on cell mitotic activity, nuclear abnormalities in interphase, extent of cell vacuole dystrophy and cell viability. RESULTS: Vaccine strains of the VEE and mumps viruses induced definite cytolytic effect in all studied cell lines, whereas the influenza virus showed such ability only in the Hep-2 cell line. Moreover, the VEE virus induced cytostatic effect in Hep-2, K-562, P3HR-1 cell lines; the mumps virus in Hep-2 and P3HR-1; and the influenza virus in Hep-2 and K- 562 cell lines. CONCLUSION: Vaccine strains of both the VEE and mumps viruses were able to induce definite cytolytic effect in all studied cell lines. However, further investigations are required to develop novel immunological methods for human tumor treatment. PMID- 17415868 TI - Neoplastic diseases prevalence in a turkish university hospital. AB - PURPOSE: The purpose of this study was to register the different cancer cases diagnosed in our hospital with an aim to define the most common and treatable cancer types and help define accurate targets for the allocation of the already limited resources of the Ministry of Health. MATERIALS AND METHODS: We surveyed 12 months of the hospital's pathology records to determine the prevalence of various cancer types. RESULTS: Out of 9720 biopsy and cytology samples, 662 were cancer cases. Breast and gastric cancers were higher and colorectal cancers were lower than the series reported from the United States. Of the pulmonary malignancies, lung cancer in general was not particularly higher in proportion to other cases, something interesting for a country with smoking rates exceeding 60% of the adult population. Squamous cell lung cancer was more common compared to the rates reported in the western world statistics. CONCLUSION: Although biases may exist, as certain cancers are more amenable to surgical intervention and physician groups may have special interest toward a particular cancer, distribution of cancer cases in Turkey is probably similar to the western world. PMID- 17415869 TI - Epidemiological prognosis of cervical cancer in Bulgaria for the period 1997 2010. AB - PURPOSE: The aim of this study was to make a prognosis for the expected levels of the basic epidemiological indicators (incidence and mortality) of cervical cancer for the period 1997 through 2010, based on the surveillance data and the disease trends obtained during the screening activities performed so far (1970-1996). MATERIALS AND METHODS: The data we studied came from the National Cancer Registry and the Central Institute of Statistics. The statistical processing of all data was performed using the software package SPSS / + v.501 for Windows and the prognosis for the expected levels of incidence and mortality of invasive cervical cancer, and incidence of carcinoma in situ (CIS) were derived through extrapolation analysis. RESULTS: Based on these data, and on the assumption that all factors affecting the incidence and mortality rates will preserve their intensity and the direction of their impact, a fairly unfavourable prognosis of increase of these indicators was obtained, with levels of about 23.7 and 9.3 per 100,000 women yearly for incidence and mortality, respectively, at the end of the prognostic period, i.e. in 2010. For comparison, the actual incidence and mortality at the start (1970) of the screening program in our country were 12.1 and 3.9 per 100,000/year women, respectively. On the contrary, further decline of the CIS incidence rate (ranging between 3.1 to 6.6 per 100,000 women for the period 1970- 1996) is expected for the period 1997-2010, and the predicted level of this indicator at the end of the prognostic period was 2.15 per 100,000 women yearly. CONCLUSION: Based on the results of this study we conclude that an aggravating unfavourable tendency of both incidence and mortality for invasive cancer and of incidence for CIS can be expected for the period 1997-2010 should the situation of lack of cervical screening programs persists. PMID- 17415870 TI - Field size and depth dependence of wedge factor for internal wedge of dual energy linear accelerator. AB - PURPOSE: To investigate the field size and depth dependence of wedge factors for internal wedge in the isocentrical setup for 6 and 15 MV photon beams. MATERIALS AND METHODS: Measurements were performed at d(max) 5, 10, and 15 cm depths for fields 5x5 cm(2) to 20x20 cm(2). The source-isocenter distance (SID) was 100 cm. Relative wedge factors (RWFs) were calculated to describe the dependence of the wedge factor on the field size and depth. RESULTS: The dependence of wedge factors on field size and depth were less than 2% at 6 MV and 1% at 15 MV Xray beams for 15 degrees and 30 degrees wedges. When the field size increased from 5x5 to 20x20 cm(2), the field size dependence increased by 2.7% and 5.2% for 45 degrees and 60 degrees wedge, respectively, at 6 MV. The dependence on the field size was less than 2.5% for 45 degrees and 60 degrees wedges at 15 MV. The depth dependence of 60 degrees wedge was up to 6.5% for large field size at the depths at 6 MV. For 15 MV beam, RWFs showed only a small change with increasing depth. CONCLUSION: The magnitude of error in using a single wedge factor was minimal for thin wedges. However, for 6 MV use of a single wedge factor produced errors up to 6% with 60 degrees wedge filter. Therefore, in the presence of 45 degrees and 60 degrees wedges in the beam, a field size and depth dependence wedge factor or a separate output factor (OF) curve for wedged beams should be used. PMID- 17415871 TI - Radiotherapy of craniopharyngioma in childhood; single institution experience. AB - PURPOSE: To present the effect of radiotherapy after subtotal excision of craniopharyngioma in childhood and the problems associated with the side-effects after radiotherapy of that rare benign tumor in children. PATIENTS AND METHODS: The study included 8 children with craniopharyngioma, treated with subtotal excision followed by radiotherapy. The median period of observation was 4 years. Radiotherapy was delivered with Co60 telegammatherapeutic machine (Rokus), with daily radiation dose (DRD) of 1.5-1.8 Gy to a total radiation dose (TRD) of 50-56 Gy. RESULTS: All 8 treated children achieved local tumor control (LTC) which was verified with dynamic observations by computed tomography (CT) and magnetic resonance imaging (MRI). In 50% of the cases the eye changes (vision, papillae, perimetric vision) were normalized, and in the remainder an improvement was noted. No child developed neurological or psychical disturbances and no memory or concentration problems ever appeared. From them 50% are very good students. CONCLUSION: Radiotherapy is the main method for achieving a long-term LTC after subtotal excision for craniopharyngioma in childhood. The radiation dose used was well tolerated by brain structures and no late neurological sequelae were observed in our patients. PMID- 17415872 TI - Philadelphia variant, t(5;9;22)(q13;q34;q11), in a case with chronic myeloid leukemia. AB - In this paper we report on a case of chronic myeloid leukemia (CML) with a Philadelphia variant involving chromosome 5 as a second change of the standard Philadelphia chromosome (Ph). Molecular analysis found a fusion gene BCR-ABL with participation of exons b3 and a2, respectively (b3a2). The molecular variant of the fusion gene BCR/ ABL or the rare involvement of chromosome 5 could possibly explain the mild course of the disease. PMID- 17415873 TI - Two cases of radiation-induced cancers 20 years after treatment of first malignancies. AB - Studies on radiation-induced breast and thyroid cancers have demonstrated that age at exposure is a major determinant of risk, with the greatest risk being for those irradiated as children and adolescents. In measuring the outcome of treatment, the ability to provide toxicity-free survival and minimal risk of second cancer may in future be regarded as important as achieving disease-free survival or local control. Herein we present two cases of secondary cancer with good treatment outcome that occurred 20 years after treatment for primary malignancies in childhood. PMID- 17415874 TI - Myeloma metastasis to the testis: findings on gray-scale and color Doppler ultrasonography with literature review. AB - Testicular metastasis of multiple myeloma is very rare and only 49 cases have been reported in the world literature. We report on a case of a 59-year-old man with a 9- month history of multiple myeloma who presented with painless swelling of the left testis. An ultrasound (US) examination was performed and the diagnosis was documented histologically. The US findings and the differential diagnosis are discussed in the light of color Doppler ultrasonography (CDUS). PMID- 17415875 TI - Gustave-Samuel Roussy (1874-1948): founder of the homonymous Institute of Cancer. AB - We present briefly through this paper the main stages of life and career of French professor Gustave-Samuel Roussy (1874-1948) as well as his efforts for the foundation of the homonymous Institute of Cancer. PMID- 17415876 TI - Solubility studies of oleanolic acid and betulinic acid in aqueous solutions and plant extracts of Viscum album L. AB - Mistletoe (Viscum album L.) contains the triterpene acids oleanolic acid (OA) and betulinic acid (BA), which were found to have anti-tumour properties. In this study, the solubilities of OA and BA were studied in water (up to 0.02 microg/mL in each case) and in alkaline solutions of 10 mM trisodium phosphate (pH 11.5; OA: 77.2 microg/mL; BA: 40.1 microg/mL). Furthermore, triterpene acids were quantified in aqueous mistletoe extracts (pH 7.3; drug to extract ratio 1:25). OA (1.1 microg/mL) and BA (0.9 microg/mL) were extracted with a yield of less than 5%. Preparing plant extracts with basic pH values resulted in a triterpene acid content of 9.3 microg/mL OA and 5.2 microg/mL BA (pH 12.1), reaching neither the solubility limits nor a complete extraction of the plant material. The triterpene acid content of neutral plant extracts above the solubility limit could be due to interactions with biocolloids. Interaction studies were performed by gel permeation chromatography. Different mechanisms of the dissolution at pH 7.3 and pH 10.2 are discussed. PMID- 17415877 TI - Resveratrol/phloroglucinol glycosides from the roots of Lysidice rhodostegia. AB - Two new phloroglucinol glycosides, lysidisides C (1) and D (2), together with two new resveratrol glycosides, lysidisides E (3) and F (4), were isolated from the n BuOH extract of the roots of Lysidice rhodostegia. The structures were elucidated on the basis of spectroscopic and chemical evidence. The antioxidant activity of the isolates was also investigated PMID- 17415878 TI - Alkaloids from Stemona mairei. PMID- 17415879 TI - Inhibition of bone resorption in cultures of mouse calvariae by apicularen A. AB - Apicularens A and B were isolated from the myxobacterial genus Chondromyces apiculatus JW184. Apicularen A inhibited bafilomycin A1-sensitive ATP-dependent proton transport into microsome vesicles more potently than apicularen B. Bone resorption in cultures of mouse calvariae induced by human parathyroid hormone (PTH) or interleukin-1beta (IL-1beta) was inhibited by apicularen A at 10 and 100 nM, while apicularen B had no effect. The bisphosphonate incadronate inhibited bone resorption at 100 nM, being less effective than apicularen A. Our findings indicate that apicularen A inhibits bone resorption induced by PTH or IL-1beta more potently than apicularen B, probably due to inhibition of the V-ATPase. PMID- 17415880 TI - Inhibitory activities of lichen-derived compounds against methicillin- and multidrug-resistant Staphylococcus aureus. AB - The inhibitory effects of selected phenolic lichen substances were tested against a panel of methicillin- and multidrug-resistant Staphylococcus aureus. Depsidones with long alkyl chains on both of the aromatic rings were consistently active against the strains tested, comparable to or better than the level of clinically used antibacterial drugs. A similar level of activity was also observed for rhizocarpic acid. The previously described cytotoxic pentacyclic compound hybocarpone was by far the most active, exhibiting minimum inhibitory concentrations (MICs) of 4-8 microg/mL (8.13-16.3 microM) against a range of multidrug efflux pump expressing strains of S. aureus. PMID- 17415881 TI - Crossing barriers in transplantation. AB - In 1978, Jonathan Sprent and Robert Korngold proved that graft-versus-host disease (GVHD) is caused by donor T cells that attack the host's non-MHC antigens. T cell depletion of donor grafts has since become a staple of transplantation strategies to combat leukemia and other inherited blood disorders. PMID- 17415882 TI - The immature spine. Proceedings of a symposium organized by the Shriners Hospital for Children, the Scoliosis Research Society, and the Pediatric Orthopaedic Society of North America. October 26, 2005. PMID- 17415883 TI - Retinal degenerative diseases. Proceedings of the XI International Symposium on Retinal Degeneration (RD2004). August 23-28, 2004. Perth, Australia. PMID- 17415884 TI - Readers' responses to "is national health insurance coming to the United States?". PMID- 17415885 TI - Proceedings of the 9th International Conference on Medical Image Computing and Computer-Assisted Intervention, MICCAI 2006. Part I. October 1-6, 2006. Copenhagen, Denmark. PMID- 17415886 TI - Readers' responses and authors' reply to "medical marijuana: politics trumps science at the FDA". PMID- 17415887 TI - Readers' responses and authors' reply to "medical marijuana: politics trumps science at the FDA". PMID- 17415888 TI - Readers' responses and authors' reply to "medical marijuana: politics trumps science at the FDA". PMID- 17415889 TI - Readers' responses and authors' reply to "medical marijuana: politics trumps science at the FDA". PMID- 17415890 TI - Proceedings of the 9th International Conference on Medical Image Computing and Computer-Assisted Intervention, MICCAI 2006. Part II. October 1-6, 2006. Copenhagen, Denmark. PMID- 17415891 TI - Proceedings of the 19th International Conference on Information Processing in Medical Imaging, IPMI 2005. July 10-15, 2005. Glenwood Springs, Colorado, USA. PMID- 17415892 TI - Readers' and author's responses to "ethics and anomalous (alternative) medical practices". PMID- 17415893 TI - Readers' and author's responses to "ethics and anomalous (alternative) medical practices". PMID- 17415894 TI - Medicalising domestic violence. PMID- 17415895 TI - Proceedings of the Third International Symposium on Gene and Stem Cell Therapy for Heart Failure and Other Cardiovascular Diseases. PMID- 17415896 TI - [Risk classification in community-acquired pneumonia]. PMID- 17415897 TI - Diagnosis of ventilator-associated pneumonia. PMID- 17415898 TI - Diagnosis of ventilator-associated pneumonia. PMID- 17415899 TI - Translating proteomics from bench to bedside. Abstracts from the Human Proteome Organisation (HUPO) 5th Annual World Congress. October 28-November 1, 2006. Long Beach, California. PMID- 17415900 TI - Biomarkers for prediction of cardiovascular events. PMID- 17415901 TI - Biomarkers for prediction of cardiovascular events. PMID- 17415902 TI - Biomarkers for prediction of cardiovascular events. PMID- 17415903 TI - Biomarkers for prediction of cardiovascular events. PMID- 17415904 TI - Biomarkers for prediction of cardiovascular events. PMID- 17415905 TI - Re: Reconstruction of the male urethra in strictures. PMID- 17415906 TI - Door-to-balloon time in acute myocardial infarction. PMID- 17415907 TI - Door-to-balloon time in acute myocardial infarction. PMID- 17415908 TI - Door-to-balloon time in acute myocardial infarction. PMID- 17415909 TI - Door-to-balloon time in acute myocardial infarction. PMID- 17415910 TI - Re: Should we screen for bladder cancer in a high-risk population? A cost per life-year saved analysis. PMID- 17415911 TI - Re: Nomograms provide improved accuracy for predicting survival after radical cystectomy. PMID- 17415912 TI - Re: Nephroureterectomy for treating upper urinary tract transitional cell carcinoma. time to change the treatment paradigm? PMID- 17415913 TI - Re: Poorly differentiated prostate cancer treated with radical prostatectomy: long-term outcome and incidence of pathological downgrading. PMID- 17415914 TI - Whipple's disease. PMID- 17415915 TI - Hemothorax after percutaneous cryoablation of the kidney: part 2. PMID- 17415917 TI - Evidence-based practice: what is it? PMID- 17415918 TI - Langerhans cell histiocytosis of the skull on cytologic squash preparations. AB - We present a case in which a primary cytodiagnosis of Langerhans cell histiocytosis (LCH) of the skull was made using squash preparations. The patient, a 25-year-old male, presented with raised intracranial pressure and decreased visual acuity. Magnetic resonance imaging revealed a large skull lesion with osteolytic features in the left frontal bone. The patient underwent surgical resection by the extended basal frontal epidural approach. The squash preparation smears were cellular and demonstrated a mixed population of small, mature lymphocytes, eosinophils, and a high histiocytes content. The histiocytes occurred as isolated or loosely cohesive and clustered. They possessed abundant cytoplasm with rounded cell shape and had characteristic nuclear features, composed of fine chromatin and delicate nuclear membranes. The cytologic features of these histiocytes were consistent with Langerhans cells (LCs). A final impression of LCH of the skull was rendered. Subsequent histopathology confirmed the diagnosis. LCs reacted with both S-100 protein and CD1a immunohistochemically. The demonstration of Birbeck granules on electron microscopic study was also noted. Whenever squash preparation yields a mixed population of mature lymphocytes, eosinophils, and histiocytes, the cytologists should be aware of and consider LCH as a diagnostic possibility. PMID- 17415919 TI - Clear cell and eosinophilic oncocytomas of salivary gland: cytological variants or parallels? AB - Oncocytomas are uncommon tumors of the salivary gland. They have an abundance of mitochondria, which is manifested as granular eosinophilic cytoplasm by light microscopy. On histological sections, presence of cytoplasmic glycogen and/or fixation artifact can impart cytoplasmic clearing, and oncocytomas with a predominance of clear cytoplasm are labeled clear cell oncocytomas. Two forms of oncocytoma, eosinophilic and clear cell, have been described in the surgical pathology literature. The purpose of this manuscript is to conduct a comparative cytological assessment to ascertain parallels and differences between the two variants. PMID- 17415920 TI - Cardiac angiosarcoma: report of a case diagnosed by echocardiographic-guided fine needle aspiration. AB - The authors present a case of cardiac angiosarcoma diagnosed by fine-needle aspiration (FNA) in a 33-year-old male. A percutaneous right atrial mass biopsy was performed via a right internal jugular venous approach and under echocardiographic guidance. FNA showed pleomorphic spindle-shaped cells present singly and in small clusters in a background of hemorrhage. Occasional pseudo acini or microacinar structures were observed, that are lined with abnormal spindly to polygonal cells, indicative of vasoformative structures. Immunostain for CD31 was strongly positive. A diagnosis of cardiac angiosarcoma was made, following which, the patient underwent treatment with chemotherapy. PMID- 17415921 TI - Esophageal leiomyosarcoma diagnosed by endoscopic ultrasound-guided fine-needle aspiration. AB - Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) has proven itself to be a reliable method for the diagnosis of gastrointestinal stromal tumors and other gastrointestinal mesenchymal neoplasms. The diagnosis and distinction of these entities remain important, as these neoplasms have different prognoses and treatment options. We present a case of a 14.5-cm esophageal mass in a 67-yr-old woman sampled by EUS-FNA. Smears showed a cellular, spindle cell lesion with marked cytologic and nuclear pleomorphism. A cell block showed neoplastic cells reacting strongly with antibodies to smooth muscle antigens and not with antibodies to keratin, CD34 and CD117. Resection confirmed the diagnosis of esophageal leiomyosarcoma. To the best of our knowledge, this represents the first reported case of an esophageal leiomyosarcoma diagnosed by EUS-FNA. Clinical, imaging, and cytologic features as well as differential diagnosis are discussed. PMID- 17415922 TI - Invasive ductal carcinoma of the breast with signet-ring cell and mucinous carcinoma components: diagnostic utility of immunocytochemistry of signet-ring cells in aspiration cytology materials. AB - Breast cancer with signet-ring cells is very rare. In this article, we present a case of invasive ductal carcinoma with signet-ring cells and mucinous carcinoma components, which could be diagnosed as a primary cancer by immunocytochemical study of fine-needle aspiration cytology material. A 73-yr-old Japanese woman noticed a palpable mass in the left breast. The aspiration cytology disclosed the monotonous proliferation of signet-ring cell with cytological atypia. The immunocytochemical examination of neoplastic cells showed a positive reaction for estrogen receptor. The extensive examination of body before the operation did not disclose any tumors in other anatomic sites. The histological examination of surgically resected breast tumor showed invasive ductal carcinoma with foci of signet-ring cell and mucinous carcinomas. Finally, our preliminary report suggests that immunocytochemical examination of aspiration cytology materials may provide useful information in searching the primary site of breast carcinoma with signet-ring cells. PMID- 17415923 TI - Fine-needle aspiration of malignant mesothelioma with unusual morphologic features: a case report. AB - Malignant mesothelioma is an aggressive neoplasm linked to asbestos exposure. Most mesothelioma patients present with pleural effusion and the fluid is typically sent for cytological examination. Therefore, cytopathologists are most familiar with features of mesothelioma in fluid preparations. We present here a case of malignant mesothelioma with unusual cytological features diagnosed on FNA. The diagnosis was confirmed by immuno-histochemical and electron microscopic studies. In addition, we compare the cytomorphological features observed in malignant effusion versus fine-needle aspiration. PMID- 17415924 TI - Readers' and author's responses to "body contouring after bariatric surgery is critical for optimal cosmetic results". PMID- 17415925 TI - Readers and authors respond to "introducing the doctor of nursing practice". PMID- 17415926 TI - Reader responds to "could computerization harm patient safety?". PMID- 17415927 TI - Readers' and author's responses to "information on complementary and alternative medicine in US government databases is biased". PMID- 17415928 TI - Readers respond to "resting metabolic rate: a critical, primary care screening test". PMID- 17415929 TI - Reader's response to "aiming high: 10 steps to a high-performance health system". PMID- 17415930 TI - Readers respond to "the $1.6 trillion question: if we're spending so much on healthcare, why so little improvement in quality?". PMID- 17415931 TI - Reader's response and author's reply to "the unexpected death of a patient in the clinical setting: some ethical reflections". PMID- 17415932 TI - Readers' and author's responses to "fixing the US physician shortage requires many more slots for resident physicians in training". PMID- 17415933 TI - Reader's response to "providing health insurance for all: what we can learn from Massachusetts". PMID- 17415934 TI - Readers respond to "balancing evidence-based medicine and cultural competence in the quest to end healthcare disparities". PMID- 17415935 TI - Reader's response and author's reply to "personalized medicine: finding the patient's 'doctor within'". PMID- 17415936 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17415937 TI - Protest and resentment. PMID- 17415938 TI - Reader's response to author's reply to "readers' response to preventing childhood obesity: an open letter to the US Congress". PMID- 17415939 TI - Findings of scientific misconduct. PMID- 17415940 TI - Bibliography. Current world literature. Valvular heart disease. PMID- 17415941 TI - Bibliography. Current world literature. Heart transplantation. PMID- 17415942 TI - Fokko Johannes van der Woude, 1953-2006. PMID- 17415943 TI - Linguistic issues and literacy barriers in nutrition. PMID- 17415945 TI - Nomenclature for two intravenous solutions. AB - May I comment on the names applied to two solutions commonly used in critical care practice, 0.9% sodium chloride (NaCl) and 5.0% glucose? I am highly aware that the pinpricking remarks following are trifling in the context of the clinical importance of the articles referred to; I am simply trying to write about scientifically correct and/or consistent use of expressions. PMID- 17415946 TI - Finding pneumo: purulent pericarditis presenting with pulsus paradoxus. PMID- 17415947 TI - Negative-pressure pulmonary oedema with normal concentration of B-type natriuretic peptide. PMID- 17415948 TI - Leptospirosis: an unusual presentation. PMID- 17415949 TI - Theoretically speaking. David Clary talks molecular quantum mechanics with Hilary Crichton. PMID- 17415950 TI - Striking a balance between the good and bad effects of UV. An up side to ozone depletion? PMID- 17415951 TI - Anthropometric measurements and dietary habits of schoolchildren in Qatar. AB - INTRODUCTION: Very little is known about the anthropometric measurements and dietary habits of children in the Arabian Gulf in general, and in Qatar in particular. OBJECTIVES: To conduct anthropometrics measurements on schoolchildren in Qatar and to explore their dietary habits. METHODS: A cross-sectional study was carried out, targeting 271 (124 male and 147 female) primary school students. RESULTS: Using the body mass index as the index of adiposity, 54.8% and 23.1% of Grade One male and female children were underweight, 3.2% and 8.8% were overweight, and 1.6% and 5.4% were obese, respectively. Breakfast, lunch, and dinner were eaten by 65%, 86%, and 87% on a daily basis. Anthropometric measurements differed significantly (P < 0.05) between genders and the other children of the National Center for Health Statistics population. CONCLUSION: schoolchildren in Qatar may be considered as having less overweight and obesity occurrence than most children in the Gulf. PMID- 17415952 TI - Antinutritional factor content and hydrochloric acid extractability of minerals in pearl millet cultivars as affected by germination. AB - Four pearl millet cultivars of two different species--Kordofani and Ugandi (Pennisetum typhoideum) and Madelkawaya and Shambat (Pennisetum glaucum)--were germinated for 6 days. The germinated grains were dried and milled. Phytic acid and polyphenol contents and hydrochloric acid (HCl) extractability of minerals from the malt flours were determined at intervals of 2 days during germination. Phytic acid and polyphenol contents decreased significantly (P <0.01) with an increase in germination time, with a concomitant increase in HCl extractable minerals. However, the major mineral content was significantly decreased while that of trace minerals was increased with germination time. When the grains were germinated for 6 days, Madelkawaya had higher extractable calcium while Ugandi had higher extractable phosphorus, whereas iron and manganese recorded high levels in Shambat and Madelkawaya, respectively. There was good correlation between antinutritional factors reduction and the increment in extractable minerals with germination time. PMID- 17415953 TI - Total phenol, antioxidant and free radical scavenging activities of some medicinal plants. AB - Phenols, a major group of antioxidant phytochemicals, have profound importance due to their biological and free radical scavenging activities. To identify their potential sources, extracts of some plants were studied for their total phenolic content (TPC), antioxidant (AOA) and free radical scavenging activities (FRSA) by different methods at multiple concentrations followed by specific phenolic composition. The amount of TPC varied from 2.8 mg/g (Withania somnifera, roots) to 107.8 mg/g (Cassia fistula, fruits) and the AOA from 24.2% (Curcuma zeoderia, leaves) to 96.9% (Trewia nudiflora, leaves). Bark of Azadirachta indica, fruits of C. fistula, and leaves and fruits of T. nudiflora were found to have high TPC (89.8- 107.8 mg/g) and high AOA (84.8-96.9%). Promising plant parts were studied for their FRSA and reducing power (RP), where the bark of A. indica, Casuarina equisetifolia and Cinnamomum zeylanicum, flowers of Indigofera tinctoria, fruits of Lawsonia inermis, and fruits and leaves of T. nudiflora showed a very low inhibitory concentration value ranging from 0.14 to 0.26 mg/ml, efficiency concentration value from 6.1 to 11.6 mg/mg DPPH and reducing power value from 0.6 to 2.8 ascorbic acid equivalents (ASE/ml), and reasonably high values (8.5 -16.2) of anti-radical power (ARP), indicating their strong FRSA. They also showed better inhibition of hydroxyl radical induced deoxyribose degradation than that of reference standard. Fruits of C. fistula with high phenols (107.8 mg/g) showed poor reducing power (5.9 ASE/ml) and ARP (4.7); in contrast, the bark of C. equisetifolia and fruits of L. inermis were with comparatively lower phenols (72.1 and 75.8 mg/g) but exhibited good ARP (16.2 and 14.4) and reducing power (0.7 and 0.6 ASE/ ml, respectively). Some of the plants were also found effective in protecting plasmid DNA nicking induced by hydroxyl radicals generated by Fenton's reaction. They were further assayed for their specific phenolic composition through high-performance liquid chromatography and MS/MS, where the amount of caffeic acid varied from 0.312 to 0.797 mg/g, chlorogenic acid from 0.018 to 2.109 mg/g, ellagic acid from 0.009 to 0.902 mg/g, ferulic acid from 0.036 to 0.078 mg/g, gallic acid from 0.192 to 3.597 mg/g, kaempferol from 0.011 to 0.910 mg/g, quercetin from 0.047 to 1.106 mg/g and rutin from 0.059 to 2.029 mg/g. PMID- 17415954 TI - A food-group based algorithm to predict non-heme iron absorption. AB - OBJECTIVE: To develop an algorithm to predict the percentage non-heme iron absorption based on the foods contained in a meal (wholemeal cereal, tea, cheese, etc.). Existing algorithms use food constituents (phytate, polyphenols, calcium, etc.), which can be difficult to obtain. DESIGN: A meta-analysis of published studies using erythrocyte incorporation of radio-isotopic iron to measure non heme iron absorption. METHODS: A database was compiled and foods were categorized into food groups likely to modify non-heme iron absorption. Absorption data were then adjusted to a common iron status and a weighted multiple regression was performed. RESULTS: Data from 53 research papers (3,942 individual meals) were used to produce an algorithm to predict non-heme iron absorption (R(2) =0.22, P < 0.0001). CONCLUSIONS: The percentage non-heme iron absorption can be predicted from information on the types of foods contained in a meal with similar efficacy to that of food-constituent-based algorithms (R(2) = 0.16, P= 0.0001). PMID- 17415955 TI - Induction of apoptosis by immature fruits of Prunus salicina Lindl. cv. Soldam in MDA-MB-231 human breast cancer cells. AB - The goal of this study was to evaluate the anticancer effect of Prunus salicina Lindl. cv. Soldam at three maturity stages (immature, midmature and mature stages). Previous studies have shown that this fruit (plums) possesses hematopoiesis effects, prevents osteoporosis and has anti-mutagenic effects. An acetone extract of immature P. salicina Lindl. cv. Soldam fruit contained higher levels of total phenolics and condensed tannins than midmature and mature plums. The results showed that an acetone extract of immature plums possesses cytotoxic effects, which are related to the activity of the total polyphenols in the fruits. Apoptosis in MDA-MB-231 cells mediated by the immature plums was associated with an increase in Bax levels and a reduction in Bcl-2 levels and the cleavage of caspase 3, caspase 7, caspase 9 and poly-(ADP-ribose) polymerase. These results indicate that immature fruit of P . salicina Lindl. cv. Soldam can be regarded as a safe and promising new dietary source for decreasing the risk of developing breast cancer. PMID- 17415956 TI - Comparative studies on storage stability of ferrous iron in whole wheat flour and flat bread (naan). AB - Whole wheat flour was fortified with premix containing ferrous sulfate, ethylenediamine tetraacetic acid and folic acid (20.0:20.0:1.5 ppm) and was stored at ambient temperature for 60 days. Naans (flat bread) were prepared from 0 ppm, 25 ppm, 50 ppm and 75 ppm ferrous iron-fortified flour samples at 10-day intervals and were analyzed for physicochemical constants and sensory evaluation. It was observed that flour containing 75 ppm ferrous sulfate contained the highest iron residues. The total iron in flour samples showed no significant difference, while ferrous iron significantly decreased in fortified flour (0.53 3.08%) and in the naans (0.42-3.48%) because of its oxidation to ferric iron during storage. The phytic acid content decreased (0.886-0.810%) significantly during the same storage period. Iron levels affected some sensory characteristics significantly (P < or =0.05), including color, texture, flexibility, chewability and overall acceptability of the naans, but not taste and flavor. The sensory attributes of naans illustrated that naans containing 50 ppm ferrous iron are more acceptable than those prepared with 75 ppm ferrous iron. PMID- 17415957 TI - Influence of the technological know-how of producers on the biochemical characteristics of red sorghum malt from small scale production units in Ouagadougou (Burkina Faso). AB - The process of red sorghum malt production was monitored three times in five production units (PU) selected upon their ability to produce malt flours having a high capacity to fluidify high-energy-density gruels. Raw, germinated and degermed seeds were analysed for macronutrient, soluble sugars, phytate and cyanide contents and alpha-amylase activity. Know-how differences between producers lay mainly in the duration and type of equipment used for steeping and germination. Moreover, three PUs applied a maturation step before sun-drying and one PU added ashes to steeped seeds before germination. No significant difference was detected in the proximate composition of malts from the five PUs. For all PUs, traditional malting increased the protein content and decreased the lipid and ash contents, while the fibre content was not affected. Significant increases in sugar contents and in alpha-amylase activity were observed but in variable proportions from one PU to another. The phytate content decreased significantly in all PUs. The cyanide content increased in all PUs but more drastically or less drastically according to the PU. Finally, degerming lowered the cyanide content to an acceptable level for human consumption. The between-PU variability may be due either to the nature and origin of the raw seeds or to technological know-how differences between producers. Further investigations are needed to optimize and standardize the malting process with a view to maximizing alpha-amylase and phytase activities and minimizing the variability of their biochemical characteristics. PMID- 17415958 TI - Antioxidant capacity, total phenolics and sugar content of selected Thai health beverages. AB - This study was to determine antioxidant capacity, total phenolics and sugar content of 12 pasteurized and sterilized Thai health beverages, products of The Royal Chitralada Projects. The antioxidant capacities were analyzed using 2,2 diphenyl-l-picrylhydrazyl radical scavenging and the photochemiluminescence (PCL) assay. Folin-Ciocalteu assay and Nelson's reducing sugar test were used to determine total phenolic compounds and sugar contents, respectively. Sacred lotus root drink showed the significantly highest antioxidant capacity in both equivalents to trolox and equivalents to ascorbic acid but not in the PCL test. In contrast, chrysanthemum drink and roselle drink showed the significantly highest values of both the total antioxidant capacity of water and lipid-soluble substances in the PCL assay. Bael fruit drink had the significantly highest total phenolic compounds. There were significant correlations between the total phenolic compounds and the antioxidant capacity values of both assays (r = 0.4 0.5). PMID- 17415959 TI - [Pay attention to the study on the role of orbital fatty tissue in thyroid associated ophthalmopathy]. AB - Increasing orbital fatty tissue volume and fatty cell number in patients with thyroid associated ophthalmopathy (TAO) can induce high orbital pressure, eyeball protrusion and visual acuity damage. Orbital fatty tissue as a new endocrine organ can secrete various fatty cell factors, growth factors and protein molecules. Some factors and protein molecules participate in the occurrence and progress of TAO. Therefore, we should pay high attention to study the important role of orbital fatty tissue in TAO. PMID- 17415960 TI - [Choice the appropriate therapy according to the clinical features of the thyroid associated ophthalmopathy]. AB - The most common etiology of unilateral and bilateral exophthalmos is thyroid associated ophthalmopathy(TAO). The diagnosis is not difficult, but the treatment is still controversy. It is the key how to recognize the clinical presentation correctly, and to select suitable therapy. PMID- 17415961 TI - [The application of botulinum toxin A in the treatment of restrictive strabismus in thyroid associated ophthalmopathy]. AB - OBJECTIVE: To observe the clinical effects of botulinum toxin A in the treatment of restrictive strabismus in thyroid associated ophthalmopathy (TAO). METHODS: Clinical data in 33 patients with restrictive strabismus in TAO treated with botulinum toxin A were analyzed retrospectively. Botulinum toxin A was injected into extraocular muscles guided by electromyography. Clinical data included the eye position, the eye movement and diplopia. RESULTS: There were 25 cases of vertical strabismus, 3 horizontal strabismus and 5 vertical combining horizontal strabismus in this group. The average degree of horizontal deviation was 35. 00+/ 20. 53 (20 - 80 , M = 27. 59 ) , the average degree of vertical deviation was 29. 33+/-17. 27 ( 10 - 100 , M = 27. 5 ) . The average follow-up time was ( 17. 04+/ 12. 77) months (5. 00+/-67. 73 months). Sixty one extraocular muscles were injected, including 31 inferior rectus, 16 medial rectus, 10 superior rectus and 4 lateral rectus. Each rectus was injected 6. 48+/-2. 12 times (4 -11times, M = 6). The interval between injections averaged (2.96+/-0. 70) months (0. 50 - 26. 00 months). The deviation degrees was decreased after injections, 15 cases were cured and did not require further surgery, 12 cases were improved and 6 cases showed poor response. The dosage for each injection averaged (8. 16+/-1. 43) U. CONCLUSIONS: Botulinum toxin A injection has a good effect in the treatment of restrictive strabismus in TAO. Better results were obtained when treated earlier. Compare with paralytic strabismus and comitant strabismus, the dosage of botulinum toxin A in each injection in TAO is higher, the mean changes of degree of deviation is slighter, the interval between injections and the duration of effect is shorter. Injection dosage should increase after repeated injections. Intolerable diplopia could be eliminated in some patients, therefore, a muscle surgery could be avoided. PMID- 17415962 TI - [Clinical application of orbital scintigraphy with 99Tc(m)-octreotide in patients with thyroid associated ophthalmopathy]. AB - OBJECTIVE: To analyze the feasibility of estimation of thyroid associated ophthalmopathy (TAO) activity with orbital scintigraphy using 99Tcm-octreotide. METHODS: There were 30 cases of TAO and 11 volunteers without eye disease or thyroid disease (control group, CG). Some examinations were performed before treatment. Based on the clinical activity score (CAS) , all TAO patients were divided into 2 groups, active group (AG) and inactive group (IAG). Orbit scintigraphy study of orbit was completed within 1. 0 - 1. 5 h after intravenous injection of 740 MBq 99Tcm-octreotide. The best transverse slice for displaying eye was selected to take semi quantitative analysis. The regions of interest of right orbit (R) , left orbit (L) and occipital (0) were selected. The uptake ratios (UR) were calculated in (R + L)/20. And then, all active patients accepted immunosuppressive therapy with prednisone after 131I therapy. Six patients performed octreotide scan twice. The statistical analyses included ANOVA, Spearman regression analysis and paired t-test using SPSS 10. 0. RESULTS: 99Tcm octreotide biokinetics showed a rapid blood clearance through urinary system. The orbit appeared as a "cold area" in CG. The orbit uptake of AG had a visual orbital accumulation of 99Tcm-octreotide, but little in IAG ( URact = 1.40 +/-0. 18, URinact = 1. 15+/-0. 10, URcontr= 1. 07 +/-0. 20). There was a significant difference between IAG and AG of UR (P <0. 001). A statistically significant correlation was found between CAS and UR (URall vs. CASall r = 0. 859,P <0. 001). After 12 weeks treatment with prednisone, the signs and symptoms of inflammation had lightened, UR pretreat vs. UR posttreat(t = 4. 39, P = 0. 007) ; CAS pretreat vs. CAS posttreat ( Z = - 5. 51, P < 0. 001 ). CONCLUSIONS: Orbital scintigraphy with 99Tcm-octreotide is a method for judging activity of inflammation in TAO based on the distribution of somatostatin receptors, lymphocyte infiltration and fibroblast activating in retrobulbar tissue. There is a good correlation between CAS and UR. The scintigraphy can estimate activity of TAO and help to formulate therapy program and evaluate the therapeutic effects. PMID- 17415963 TI - [Establishment of an animal model for thyroid associated ophthalmopathy by treatment of mice with human thyrotropin receptors-activated splenocytes]. AB - OBJECTIVE: To establish an animal model for thyroid-associated ophthalmopathy (TAO) by treatment of homoimmune BALB/c mice with human thyrotropin receptors ( hTSHR )-activated splenocytes. METHODS Twenty three BALB/c mice were randomly divided into group A and B, which were immune with recombinant plasmid pcDNA3. 1/hTSHR or blank plasmid pcDNA3. 1 for 3 times at 3-week intervals, respectively. Mice in these two groups were sacrificed 18 weeks after immunization and the splenocytes were isolated. Other 26 homoimmune BALB/c mice were divided randomly into group C and group D which received splenocytes from group A and group B, respectively. Four weeks later the orbital tissues were harvested for pathological examination and serum was collected for TT4, TSH and TRAb measurements. RESULTS After immunization with recombinant plasmid pcDNA3. l/hTSHR, orbital tissues displayed edema and mucinous degeneration in 25% of mice but no proliferation of adipose tissue and fibrous tissue (group A). After immunization with hTSHR-activated splenocytes, orbital tissues displayed proliferation of adipose tissues and fibrous tissues, degeneration and disruption of muscular fibers microscopically in 50% of mice (group C). Under electronic microscope, expansion of smooth endoplasmic reticulum and disorder of myofibrils were found in these mice. In all mice from group B and group D, orbital tissues were normal histologically. After immunization with splenocytes, serum total T4 levels were significantly elevated ( P = 0. 036) in group C (7. 130+/-1.017) [microg /dl when compared with group D (6. 431+/-0. 573) microg /dl. Serum TSH levels were significantly reduced (P = 0. 020) in group C (0. 070+/-0. 032) microIU/ml when compared with group D (0. 098+/-0. 020) microIU/ml. Serum TRAb levels were slightly increased in group C(0. 202 +/-0. 067) as compared to group D(0. 151+/-0. 055) , however, this difference was statistically non-significant (P = 0. 055 ). CONCLUSIONS: TAO animal model established by immunizing homoimmune BALB/c mice with hTSHR-activated splenocytes is similar to human TAO in histological characteristics and is a feasible and reliable experimental method. Our study also proves that the thyrotropin receptor and T cell aiming directly at this auto-antigen play important roles in the autoimmune process of TAO. PMID- 17415964 TI - [Peripapillary atrophies in primary open-angle glaucoma and glaucoma-like normal disks]. AB - OBJECTIVE: To evaluate the variety of peripapillary atrophy (PPA) and its relationship with visual field among primary open angle glaucoma (POAG), normal tension glaucoma (NTG), and normal subjects with glaucoma-like disk (GLD) that had no evidence of visual field defect and retinal nerve fiber layer defect. METHODS: The study included 42 eyes of 42 patients with POAG, 40 eyes of 40 patients with NTG, and 45 eyes of normal subjects with GLD. Stereo color optic disc photographs were morphometrically evaluated by a video camera and computer system. The refractive range ( spherical equivalent) of all these subjects was between + 3.00D and -3.00D. RESULTS: The alpha-zone and beta-zone of PPA occurred significantly (P <0. 05) more frequent in both patients with POAG( 100. 0% ,48. 9% )and with NTG(95.0% ,37. 5% )than that of normal subjects with GLD(85.4% ,19. 1% ) , but no difference was found between the POAG and NTG groups (P > 0. 05). The beta-zone area of PPA was significantly ( P < 0. 05) enlarged in POAG compared with GLD normal subjects. While there is no significant difference in alpha-and beta-zone area between all other groups. In glaucoma patients, The alpha-zone or the beta-zone area of PPA weakly correlated with the glaucomatous visual field defect While cup/disk area ratio showed a relatively strong correlation. Cup/disc area ratio and age were the only two risk factors for the occurrence of glaucomatous visual field defect among all the factors including PPA by logistic regression analysis. CONCLUSIONS: Our results demonstrate that cup/disc area ratio is well correlated with glaucomatous visual field defect. The occurrence of alpha-zone, beta-zone, and beta-zone areas of PPA may not be a clinically useful parameter, for diagnosing or differentiating POAG and NTG . PMID- 17415965 TI - [DNA damage and repair induced by acute exposure of microwave from mobile phone on cultured human lens epithelial cells]. AB - OBJECTIVE: To investigate the effects of acute exposure of low-power 217 Hz modulated 1. 8 GHz microwave radiation on the DNA damage of human lens epithelial cells (hLECs) and repair. METHODS: Cultured hLECs were exposed to 217 Hz modulated 1. 8 GHz microwave radiation at SAR (specific absorption rate) of 1. 0, 2. 0, 3. O0 and 4. 0 W/kg for 2 hours in an sXc-1800 incubator and irradiate system, the DNA single strand breaks were detected with comet assay ( single-cell gel electrophoresis) in sham-irradiated cells and irradiated cells incubated for varying periods: 0, 30 and 60 minutes after irradiation. Images of comets were digitized and analyzed using an Imagine-pro plus software, and the indexes used in this study were tail length (TL) and tail moment (TM). BrdU was added into the medium with additional one hour incubation after radiation, the cell proliferation rate was determined using a BrdU-kit. RESULTS: The difference of DNA-breaks between the exposure and sham exposure groups induced by 1.0 and 2.0 W/kg irradiation were not significant in each time points (P > 0.05) ; there were significant difference in both groups at the exposure dose of 3. 0 and 4. 0 W/kg immediately and at the time of 30 minutes after irradiation (P <0. 01) ; if the radiation exposure time was beyond one hour no differences were be able to detected in 3.0 W/kg group (P > 0. 05) compared with control, but the evidence of significant DNA damage still existed in 4. 0 W/kg group at the same time point. Cell proliferation rate had no significant difference when the application of SAR was < or = 3. 0 W/kg (P >0. 05) , however the cell proliferation was decreased significantly at the dose of 4. 0 W/kg irradiation ( P < 0. 01). CONCLUSIONS: No effective DNA damage was induced using comet assay after 2 hours irradiation of 1. 8 GHz microwave on hLECs at the dose SAR < or = 3.0 W/kg. 4.0 W/kg irradiation caused significantly DNA damage and inhibition of hLECs proliferation. PMID- 17415966 TI - [Effects of human retinal glial cells under hypoxic conditions on circulating endothelial progenitor cells]. AB - OBJECTIVE: The specific signaling events by which circulating endothelial progenitor cells (EPCs) responding to alterations of the retinal vasculature induced by hypoxia are not yet completely understood. Our goal was to generate a suitable experimental model for the identification of such factors as well as the effects of these factors on the EPCs. METHODS: Primary cultures of human retinal glial cells were incubated under hypoxic or normoxic conditions. Effects of hypoxia were confirmed by analysis of HIF-lalpha and VEGF expression. Circulating EPCs (CD34 +, CD133+ double-positive cells) were isolated by FACS Vantage from the human periphery blood. EPCs were treated with conditioned media from retinal glial cells incubated under either hypoxic or normoxic conditions. Effects of conditioned media on the proliferation and differentiation of EPCs were evaluated. RESULTS: Incubation of human retinal glial cells at low oxygen concentration for 24 hours lead to HIF-lalpha expression and a significant increase of VEGF released to the media[ (319. 16 +/-34. 12) pg/ml versus (220. 28+/-24. 33) pg/ml] as compared to the control cultures incubated at normoxic conditions (P <0. 01). Treatment of EPCs with conditioned medium from retinal glial cells incubated at low oxygen concentration resulted in an increase of proliferative activity (day 3: 0. 83+/-0. 10 versus 0. 61+/-0. 22,P <0. 05; day 7: 1. 53+/-0. 21 versus 0. 88+/-0. 25, P <0. 05) and presence of differentiated endothelial cells as compared to cells cultured with conditioned media from normoxic cultures. CONCLUSIONS: Conditioned media from retinal glial cells contain factors capable of activating the proliferation and differentiation of circulating EPCs. Analysis of the effects of conditioned medium from neuronal glial cells incubated under hypoxic conditions on circulating human EPCs might be useful in providing an effective experimental method for the identification of the specific signaling molecules involved in the activation of circulating EPCs for the regeneration of retinal blood vessels. PMID- 17415967 TI - [Relationship between blue light-induced apoptosis and mitochondrial membrane potential and cytochrome C in cultured human retinal pigment epithelium cells]. AB - OBJECTIVE: To investigate the effect of blue light on apoptosis and mitochondrial permeability transition (MPT) of cultured human retinal pigment epithelium (RPE) cells in vitro. METHODS: Human RPE cells were exposed to blue light (wave length 470 -490 nm). The present study consisted of three parts. Part one studied the effect of various intensities of blue light on the RPE cells. Cells were irradiated with (500+/-100) lx (group 1) , (2000+/-500) lx (group 2) and (3000+/ 500)lx ( group 3) blue light, and followed by 24 hours observation. Part two studied the effect of various duration of blue light at identical intensity on the RPE cells. For the study on various subtypes of RPE cells, cells were irradiated by blue light at (2000+/-500) x for 6, 12, and 24 hours. For the study of mitochondrial membrane potential, cells were irradiated for 3, 6, and 12 hours. Part three studied cells irradiated with blue light at identical intensity and duration, but with various prolongation of post-exposure culture. The prolongation of post-exposure culture was 6, 12, 24, and 36 hours. Phototoxicity was quantified at various periods after exposure by staining of the nuclei of membrane-compromised cells, by TdT-dUTP terminal nick-end labeling (TUNEL) of apoptotic cells and by Annexin V labeling for phosphatidylserine exposure. Transmission electronmicroscopy was used to determine the ultrastructure changes of RPE cells. Mitochondrial membrane potential ( deltaPsim ) was measured by rhodamine 123 staining and subsequent flow cytometry. Cytochrome C activity was assayed by ELISA. Caspase-3 was detected by colorimetric assay. RESULTS: TUNEL positive labeling cells in first group of part two study showed cell shrinkage, membrane blebbing, apoptotic body, condensation and fragmentation of chromatin. Mitochondrial swelling, extinction of inner mitochondrial membrane ridge, dilation of rough endoplasmic reticulum and increase of the lysosome were also observed in transmission electronmicroscopy. Blue light at (500 +/- 100) x intensity did not induce damage to RPE cells, but decrease of delta Psim was observed. A significant increase of apoptotic, apoptotic necrotic and necrotic percentages, as well as significant decrease of deltaPsim were observed at higher light intensity in part one study. Increase of apoptotic percentage was the main response to shorter exposure of blue light. Increase of apoptotic necrotic and necrotic percentage and pronounced decrease of deltaPsim occurred in cells irradiated by longer exposure in part two study. In part 3 study, apoptotic response was increased gradually during 6 and 12 hours prolongation of post exposure culture, more apoptotic necrosis or necrosis were found after post exposure 24 hours. Decrease of deltaPsim was observed in 6 hours prolongation of post-exposure culture and lasting for 48 hours. The concentration of cytochrome C was significantly increased in post-exposure 24 and 36 hours, without any changes of Caspase-3 activity. CONCLUSIONS: Blue light exposure can induce damages to human RPE cells in vitro, which include apoptosis, apoptotic necrosis and necrosis. These changes are caused by triggering the mitochondrial permeability transition, which results in decrease of deltaPsim and release of cytochrome C. deltaPsim can be used as a earlier parameter of blue light-induced apoptosis. PMID- 17415968 TI - [Effects of magnetic field on MAPK signaling pathways of human retinal pigment epithelial cells bound with beads in vitro]. AB - OBJECTIVE: To observe the changes of mitogen activated protein kinase (MAPK) signaling pathways of human retinal pigment epithelial cells( RPEs) bound with beads under stretch caused by the force from magnetic field in vitro. METHODS: Ferric oxide microparticles, coAted with collagen, were added to dishes containing substrate-attached RPEs. After incubation, the cell layer bound with beads was laid in a magnetic field, the cells were stretched by the magnetic force. The activation status of the extracellular signal-regulated protein kinase (ERK), c-jun N-terminal kinase (JNK), and p38 in RPEs was determined over a time course from 3 to 30 minutes with western-blot analysis. To examine the role of p38 kinase in the response to stretching, cells were grown for 30 minutes in the presence or absence of inhibitor of p38 (SB203580). The changes of the expression of active p38 kinase were observed with fluorescence staining. RESULTS: Total ERK, JNK, and p38 were detected in RPEs. Active ERK and p38 were detected but active JNK was not detected. Activation of ERK was unchanged during the time course. In contrast, p38 activation was barely detected in the normal cells, but this stress-activated protein kinase exhibited a robust activation after 5 minutes in the magnetic field. SB203580 blocked the p38 activation during stretch stimulation. The stretch stimulation also increased the fluorescence staining of active p38. CONCLUSION: A magnetic field can affect RPEs bound with beads. The effect may be partially through p38 signaling pathway. PMID- 17415969 TI - [Expression of muscarinic acetylcholine receptor-1 in human retinal pigment epithelium]. AB - OBJECTIVE: The aim of this study was to identify the presence of muscarinic acetylcholine receptors-I (M1 receptor) in human retinal pigment epithelium (RPE) in order to determine the role of M1 receptor in the maintenance of function of RPE and its role in the occurrence and development of myopia. METHODS: The 3rd 5th passages of RPE cells established in our laboratory were used in the present study. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect mRNA expression of M1 receptor in cultured RPE. Immunocytochemistry was used to detect M1 receptor protein in the RPE cells. RESULTS: Cultured RPE demonstrated mRNA expression of M1 receptor in RT-PCR. Protein of M1 receptor was presented in the RPE under immunocytochemistry. CONCLUSIONS: This study demonstrated the presence of M1 receptor in human RPE at both mRNA and protein levels. M1 receptor plays an important role in the maintenance of function of RPE. Injection of M1 receptor antagonist into the vitreous can delay the occurrence and inhibit the development of myopia, which is possibly related to the inhibition of RPE cells function. PMID- 17415970 TI - [Study of genetic mutation locus in a family with congenital aniridia]. AB - OBJECTIVE: The aim of this study was to determine the genetic mutation locus of congenital aniridia. METHODS: Peripheral vein blood (2-5 ml) was collected from all members of a congenital aniridia family. DNA was extracted, then, the primers of polymorphic microsatellite genetic markers were synthesized, followed by polymerase chain reaction (PCR) analysis. Polyacrylamide gel electrophoresis (PAGE) was used to analyze the denatured PCR products and haplotype linkage analysis was applied according to the relationship between bands and family members to determine the association between the phenotype of aniridia and PAX6 gene. Fourteen exons of human PAX6 gene were amplified by PCR and allele specific variations were detected by single strand conformation polymorphism (SSCP). By comparing the difference of bands between patients and the unaffected members, the mutated exons was detected. The mutated locus was detected by direct DNA automated sequencing of the PCR products with different SSCP bands. RESULTS: PAX6 mutation was linked to the occurrence of aniridia in this family. An extra band corresponding to exon 9 in PAX6 was found by single strand conformational polymorphism analysis in all aniridia patients in this family, but not detected in the unaffected members. A mutation of C to T was detected by sequencing at the nucleotide 1080 that converts the Arg codon (CGA) to a termination codon (TGA). CONCLUSION: Mutation of PAX6 gene can result in the occurrence of congenital aniridia. PMID- 17415971 TI - [The suppression of the experimental autoimmune uveoretinitis with retinal S antigens by intranasal tolerance induction]. AB - OBJECTIVE: The aim was to investigate the suppression of rat experimental autoimmune uveoretinitis( EAU) by induced immune tolerance via intranasal administration of retinal S antigens. METHODS: The bovine S antigen was purified from bovine retina by salt precipitation and ionic exchange chromatography, the female Lewis rats were used to induce immune tolerance by intranasal administration with purified bovine retinal S antigens and then the rats were used to produce the EAU model by retinal S antigens challenge. The rate of EAU occurrence, the clinical and histological scores, the skin delayed-type hypersensitivity and lymphocyte proliferation stimulated by retinal S antigen and concanavalin A were recorded. The adjunct effect of cyclophosphamide on tolerance induction was observed. RESULTS: After intranasal administration of retinal S antigens, EAU was induced in two of eight ( 25% ) rats in tolerant group, sis of six (100%) rats in control group , the difference of EAU induction rate was significant in tolerant group compared with control (P = 0. 0097) . The average onset time in tolerant group were 16. 5 days, the control group was 10. 3 days, the difference was significant ( F = 26. 32, P = 0. 000; q = 9. 723, P <0. 01). The average clinical scores of EAU in tolerant group were 0. 89, the control group was 3. 94, the difference was significant( F = 12. 48 ,P = 0. 000; q = 7. 904, P < 0. 01 ). The average histological scores of EAU in tolerant group were 1. 21, the control group was 4. 12, the difference was significant( F = 11. 80, P = 0. 000; q = 7. 510,P <0. 01). The histological features in tolerant group were iris blood vessels slightly dilation, few exudates in anterior chamber and vitreous cavity; there were slighter retina swallow and the photoreceptors damages in the tolerant group. The skin delayed-type hypersensitivity and the proliferative responses of lymphocytes stimulated by S antigen and concanavalin A in tolerant group were slighter than that in the control group. Intraperitoneal injection of cyclophosphamide enhanced the effect of immune tolerance slightly. Only intraperitoneal injection of cyclophosphamide did not diminish the severity of the rat EAU. CONCLUSION: The intranasal induced tolerance by retinal S antigens can suppress effectively the prevalence of rat experimental autoimmune uveoretinitis induced by retinal S antigens. PMID- 17415972 TI - [Biometry of the anterior chamber diameter]. AB - Interest in the use of phakic intraocular lens for the treatment of high myopia and hyperopia is increasing. Several approaches have been advocated including angle-supported phakic intraocular lens. The implantation of anterior chamber intraocular lens is the good remedy for the cases in whom posterior chamber intraocular lens can not be implanted. However, the implantation of the anterior chamber intraocular lens has been associated with several potential complications, many of which arise from poor sizing. To reduce the incidence of these complications, accurate anterior chamber biometry is necessary; therefore we summarized the research advances of improvement of the biometry of anterior chamber diameter. PMID- 17415973 TI - [Science-based management of infectious disease crisis]. PMID- 17415974 TI - [Second thoughts on metabolic syndrome]. PMID- 17415975 TI - [Metabolic syndrome: cunent status and perspective]. PMID- 17415976 TI - [Is hypertension a component of metabolic syndrome?]. PMID- 17415977 TI - [Metabolic syndrome and prevention of cardiovascular diseases]. PMID- 17415978 TI - [Epidemiological study on injuries among children in Beijing]. AB - OBJECTIVE: To explore the morbidity, burdens caused and risk factors of injuries among children and to provide information for policy making on prevention. METHODS: By PPS sampling method, we selected 28 000 households in Beijing and using a questionnaire developed by UNICEF, TASC and China CDC. RESULTS: The morbidity of child injury in Beijing was 2.25%, higher than that of adults (1.56%) with 6-9 year group the highest (2.83% ). The morbidity of boys was 3.04% higher than that of girls (1.37%). Among the 7 death cases, 3 were caused by injury(2 RTA, 1 drowning). The three leading causes of injuries were fall, animal bite and road traffic accident in children. Among children aged 0-5 years, fall, animal and burn/machine injuries were the leading causes. Among children aged 0-5 years, the leading causes were hurt by animals, fall and RTA with the leading causes as fall, sprain and RTA. The average medical cost of injury cases was 1617.57 Yuan. There were 3500 children (injured orphan) whose parents were injured leading to death or disability in Beijing in 2003. CONCLUSION: Injury was the leading cause of child deaths in Beijing with children of 6-9 years old having the highest morbidity with RTA the most severe type of injuries in Beijing which affects both children and their parents. PMID- 17415979 TI - [The current status and the changing patterns of perinatal health care in some southern and northern areas of China, 1994 - 2000]. AB - OBJECTIVE: To understand the current status and its changing patterns of perinatal health care in some southern and northern areas of China from 1994 to 2000. METHODS: Data was collected in 13 counties/cities covered by Perinatal Health Care Surveillance System which was part of the Sino-American cooperative project on neural tube defects prevention, established in 1992. The study population consisted of 368 589 women who delivered single live births of at least 20 gestational weeks from 1994 through 2000. Chi-square test was employed to test the difference on the proportions of women with prenatal checkup, women with early prenatal care onset, women with at least five prenatal visits, hospital delivery versus home delivery as well as the proportion of women with at least three postpartum visits. Two-way ANOVA was used to test the differences of mean of gestational weeks at first prenatal visit and the mean of prenatal visits between different year and areas. RESULTS: The proportion of women with prenatal checkup remained over 99.0% from 1994 through 2000. The mean of gestational weeks at first prenatal visit changed from 13.1 weeks in 1994 to 10.7 weeks in 2000. The proportions of women with early prenatal care onset increased from 65.5% in 1994 to 79.4% in 2000 while women with at least five prenatal visit increasing from 34.1% to 71.8%, hospital delivery increasing from 91.3% to 98.8%, home delivery decreasing from 5.6 % to 0. 6%, women with at least three postpartum visits increasing from 80.5 % to 95.1%. Except for the proportion of women with early prenatal care onset, other proportions in northern areas were lower than those in southern areas in the same year. Greater differences between urban and rural areas in northern areas were found than in southern areas. CONCLUSION: The utilization of perinatal health care services was greatly improved in both southern and northern areas from 1994 to 2000. However, there were still obvious differences in the perinatal health care services between northern and southern areas as well as between urban and rural areas in northern China. PMID- 17415980 TI - [Study on physical activities status of citizens from different subpopulation in Pudong, Shanghai]. AB - OBJECTIVE: To understand the levels of total physical activities among different populations, and the distribution of four domains. METHODS: With proportional stratified sampling and cluster sampling method, 3555 subjects were selected including officials, company staff, high school students, community population and floating population. RESULTS: The proportions for inactive, insufficiently active, minimally active and health enhancing physical active (HEPA active) as a whole were 20.5%, 10.1%, 26.5% and 42.9% respectively. Community population had the highest level of HEPA active which was 65.6%. Floating population had the highest level of inactivity of 33.1%. Apart from the male floating population, the active level in transportation, physical activity among the investigated were the highest compared to other three active domains. Leisure-time physical activity was in the opposite. CONCLUSION: The type of physical activities among general citizens were mainly of physical activity related to transportation but less leisure-time physical activity was seen. Floating population had the highest level of both HEPA activity and inactivity. PMID- 17415981 TI - [Study on the current status and relative factors on reproductive health of unmarried floating population in Huizhou City]. AB - OBJECTIVE: To understand the current status and relative factors of reproductive health (RH) on unmarried floating population. METHODS: Both quantitative and qualitative surveys were employed. Quantitative survey would include questionnaire admimistration, physic examination and laboratory test while qualitative study including focus/nominal group discussion, individual and resource persons interview. All of the investigative targets were selected under informed consent. RESULTS: Among 1333 investigated unmarried floating population, the mean score on knowledge of RH was 12.97 +/- 12.20 (the full score was 100). Among studied persons, 10.20% of them were in favor of unmarried sexual activity while 50.64% of them did not care about it and 26.90% of them had ever experienced sexual activity. Among females who experienced sexual activity, 25.45% of them had induced abortion. It was found, through multivariate logistic analysis, that sexual activity at younger age, less education, agreed on having sexual activity under partner's request, sexual activity in half of a year, and pregnancy were the major risk factors on RH of unmarried floating population. CONCLUSION: The problem on the current status of RH among unmarried floating population needs to be addressed as they were lack of knowledge on RH and having more risk factors on RH. PMID- 17415982 TI - [Study on the comparison of three diagnostic criteria of metabolic syndrome in Jiangsu population]. AB - OBJECTIVE: To compare the significance of the application in Jiangsu population using the diagnostic criteria for metabolic syndrome(MS) proposed by the IDF in 2005, ATP III in 2005 and CDS in 2004. METHODS: Based on the populations in Jiangsu province from a project of Multiple Metabolic Disorders and MS, the study was conducted including 5888 cases, with data of plasma glucose, lipid profile, blood pressure, serum insulin etc. MS was diagnosed and compared according to these three definitions respectively. RESULTS: The age-adjusted MS prevalence rates were 17.48%, 21.95% and 9.59% according to the IDF(2005), ATP III (2005) and CDS (2004) respectively. The agreement in the diagnosis of MS using ATP III (2005) and CDS(2004) definitions was 85.11%, and the agreement in the diagnosis of MS using IDF(2005) and CDS definitions was 87.35%. The agreement in the diagnosis of MS using IDF (2005) and ATP III (2005) definitions was 95.14%. The MS subjects diagnosed by the ATP III (2005) was 1.26 higher than subjects diagnosed by the IDF(2005) definition. The ratios of prevalence rates of high waist circumference(WC), MS_IDF (2005) and MS_ATP III (2005) was 2.17, 1.99 and 1.54 in sex ratio (woman to man). CONCLUSION: The agreement in the diagnosis of MS using the IDF(2005) and ATP III (2005) definition was higher than using CDS(2004) and other two definitions. For diagnosing MS, the cut off of WC in IDF(2005) and ATP III (2005) seemed not appropriate and the diagnostic criteria used for ATP III (2005) (waist circumference of man 85 cm, woman 80 cm) could identify more MS. PMID- 17415983 TI - [Geography and host distribution of Crimean-Congo hemorrhagic fever in the Tarim Basin]. AB - OBJECTIVE: To determine the infective status and natural distribution of Xinjiang hemorrhagic fever (XHF; Crimean-Congo hemorrhagic fever, CCHF) in ticks, rodents and livestock in the Tarim Basin. METHODS: The pathogenic materials of ticks or rodents' viscera and blood samples of sheep were inoculated into sucking mouse of 24 to 48-hour old. Materials with typical clinic symptoms were identified with RPHA and IFA. RT-PCR was taken to detect special S gene segment of Crimean-Congo hemorrhagic fever virus (CCHFV) in the objective material. RESULTS: All the samples of ticks, rodents' viscera and blood samples of sheep from 21 counties (cities) in the Tarim Basin were divided into 422 groups and inoculated into sucking mouse at laboratory. 49 materials with typical clinic symptoms were obtained. The morbidity rate with typical clinic XHF was high in Bachu, Yuli, Yutian and Ruoqiang. There were 43 samples identified with RPHA with 6 positive samples and positive rate of 1.4%. The materials with positive RPHA were found in Yuli, Luntai and Yutian. 42 samples were identified with IFA and 13 positive samples with the positive rate of 3.1%. The positive materials of IFA were found in Bachu, Yuli, Minfeng, Luntai and Yutian. 32 samples were detected with RT-PCR and there were 31 samples with special S gene segment of CCHFV (329- 548 nt). The positive materials was widely distributed in Aksu, Awat, Bachu, Luopu, Yuli, Minfeng, Qiemo, Ruoqiang, Luntai and Yutian. The highest infective rate was in Hyalomma asiaticum kozlovi, and followed by sheep. S gene segment was detected in viscera of M. meridianus. CONCLUSION: XHF relied on the river in the southern part of Xinjiang and distributed in the areas with Populus euphratica shrub in desert and oasis in the Tarim Basin. The main vector and host were Hyalomma asiaticum kozlovi. Livestock such as sheep, camel, L. yarkandensis, M. meridianus and Euchoreutes naso could serve as the deposited host of XHF. PMID- 17415984 TI - [Rapid simultaneous detection of Salmonella and Shigella using modified molecular beacons and real-time PCR]. AB - OBJECTIVE: Dual detection of Salmonella and Shigella using modified molecular beacons and real-time PCR was developed. The established method was applied to rapid diagnosis of Salmonella and Shigella' food poisoning, and for routine monitoring programs. METHODS: Two sets of primers were designed based on the core sequence of invA gene and ssaR gene published on GenBank to detect Salmonella, and ipaH gene were selected to detect Shigella. Three corresponding modified molecular beacons labeled with different fluorophors were designed. The molecular beacons and primer sets were tested against numerous strains from 55 different bacterial species. Then the two assays were combined to establish the dual real time PCR assay, and were applied to the food poisoning diagnosis and surveillance. RESULTS: For the modified molecular beacons-based dual real-time PCR assay, the sensitivity achieved was 69-93 fg/microl, 32-64 CFU/ml or 1-2 CFU/PCR reaction. There was no cross-reaction with other bacteria served as control. The dual real-time PCR assay was used to detect 134 Salmonella strains and 67 Shigella strains but no false signals were observed. 1100 food poisoning samples were tested with 569 Salmonella and 42 were Shigella identified by real time PCR. Among the positive samples, 551 were detected Salmonella and 41 were Shigella by traditional culture method. The overall test could be finished within 2 hours to one day starting from sample preparation. CONCLUSION: The modified molecular beacons-based dual real-time PCR assay was rapid, sensitive, and specific. It could be applied to the rapid diagnosis of Salmonella and Shigella' food poisoning. PMID- 17415985 TI - [A clinical study on hepatitis B virus genotype]. AB - OBJECTIVE: To study the hepatitis B virus (HBV) genotype and its relation to clinical degree and responsiveness to antiviral therapy on hepatitis in order to guide the clinical therapy. METHODS: We amplified HBV S gene by polymerase chain reaction (PCR), using the second-round PCR product, which was digested by restriction fragment length polymorphism (RFLP). This genotype method was designed under the analysis of the restriction fragment length polymorphism and using the restriction enzymes that identified the genotype-specific sequences. Five restriction enzymes, Hph I , Nci I , Alw I, Ear I and NlaIV, were identified in genotype-specific RFLP from the S gene region. Representative sequences from the S genome region of each HBV genotype were aligned to show the restriction sites by the five restriction enzymes. The amplified S gene nucleotide sequences were sequenced by dideoxy-chain-termination method and the corresponding amino acid sequence was deduced using DNASIS software. Later, they were genotyped by comparing to representative S gene sequences obtained from GenBank. This confirmed the results of RFLP HBV genotyping methods, coincident with that of S gene sequence. RESULTS: Genotypes A, B, C, D were classified in 216 patients with HBV and DNA positive. The results showed that: 1 case (0.46%) of genotype A, 19 cases genotype B (8.8% ), 175 genotype C (81.02%) and 21 genotype D (9.72%). A total of 86 patients in the hospital were divided into either genotype C cases (69) or non-genotype C cases (17). CONCLUSION: Genotype C was the major genotype in Changchun. Among HBV patients, type C was 80.95%, followed by genotypes B and D. Both hepatocellular carcinoma and liver cirrhosis showed relations with genotype C. PMID- 17415986 TI - [Genotype identification and sequence analysis of Orientia tsutsugamushi isolated from Shandong area]. AB - OBJECTIVE: To determine genotype, nucleotide sequence homology and phylogenesis of Orientia tsutsugamushi isolated from Shandong, China. METHODS: Orientia tsutsugamushi isolated from patients, Apodemus agrarius and Leptotrombidium scutellare in Shandong area were identified by nested-PCR. On the basis of the nucleotide sequence of the gene that encoding the Ot M, 56 x 10(3) antigen, the primers were frequently used in Japan and Korea. Nucleotide sequences of three isolates were determined. The DNA sequences were compared with nucleotide sequences of Orientia tsutsugamushi registered in GenBank for sequence homology analysis. Phylogenetic analysis of the isolates and'some published sequences was carried out with Neighbor-joining method by MEGA 3.1 software. RESULTS: 481- 507 bp DNA fragments encoding Orientia tsutsugamushi M, 56 x 10(3) protein were amplified successfully in the samples of Gilliam, Karp, Kato and Shandong isolates by group-specific primers. The corresponding target fragments of the three international reference strains of Gilliam, Karp, and Kato were amplified successfully with each of their own type specific primers. 523 bp DNA fragments were amplified successfully from Shandong isolates by the nPCR with Kawasaki specific primer, and no DNA fragment was amplified by the nPCR with Gilliam, Karp, Kato, Kuroki and Saitama-specific primer. Comparing with the sequences of Orientia tsutsugamushi registered in GenBank, all the Shandong isolates shared higher than 95% nucleotide sequence homology with Kawasaki strain founded in Japan. Data from phylogenetic analysis showed that Shandong isolates belonged to the same branch with Kawasaki strain. CONCLUSION: To facilitate international comparison and communication, the primers should be employed in the Orientia tsutsugamushi research in China. Orientia tsutsugamushi isolated in China were similar to Kawasaki strain PMID- 17415987 TI - [Comparison of multilocus sequence typing system and pulsed-field gel electrophoresis in typing of Salmonella enteritidis]. AB - OBJECTIVE: To compare the discriminatory ability between multilocus sequence typing system (MLST) and pulsed-field gel electrophoresis (PFGE). METHODS: Salmonella enteritidis strains, isolated from food in China were identified by MLST, under PCR products of thrA, purE, sucA, aroC, hemD, dnaN and Sdf I. The same set of strains was typed by PFGE using Xba I, Spe I as the restriction enzyme in order to compare the discriminatory power of the methods. RESULTS: Data from MLST revealed the lack of diversity among the strains of the same serotype and the number of variable nucleotide sites per locus ranged from 1 to 23 between Salmonella typhi LT2 and other serotypes of Salmonella. However, 50 Salmonella enteritidis strains were identified as 11 patterns and more sub-patterns by PFGE. CONCLUSION: In strain typing, PFGE was the highly discriminatory method comparing to the MLST system. PMID- 17415988 TI - [Study on the differences on HA1 regions between epidemic strains and vaccine strains of influenza virus subtype A3 from 1988 to 2005]. AB - OBJECTIVE: To study the differences between epidemic strains of influenza virus subtype A3 circulated in China and Occident in past 18 years, in genetic level, and vaccine strains recommended by WHO in corresponding time. METHODS: Amino acid sequences of HA1 regions of the epidemic strains, which circulated in China and Occident from 1988 to 2005, and the vaccine strains of influenza virus subtype A3 were compared by BioEdit and analyzed on the differences of HA1 and it's antigen determinants RESULTS: Differences between epidemic strains and vaccine strains recommended in corresponding year, both in HA1 and it' s antigen determinant regions, were obviously greater than that between epidemic strains and vaccine strains recommended in next round (P< 0.01). However, differences between epidemic strains in Occident and vaccine strains recommended in corresponding year were slightly greater than that between epidemic strains and vaccine strains recommended in next round and it was not marked (P >0.05). In addition, differences between epidemic strains and vaccine strains which being used for several years, whether in China or in Occident, constantly increased accompanying the used time prolonged. CONCLUSION: There was an obvious lag between vaccine strains recommended by WHO, analyzed in genetic level, and epidemic strains of influenza virus subtype A3 circulated in China. PMID- 17415989 TI - [Study on the immunogenicity of major leptospiral genus-specific protein antigens and the distribution of antigens in different serogroups of Leptospira interrogans]. AB - OBJECTIVE: The determination of antigenicity and immunogenicity of Leptospira interrogans genus-specific outer envelope proteins (OEPs) will offer evidence for developing universal leptospiral genetic engineering vaccine and detection kit. METHODS: In this study, Ni-NTA affinity chromatography is used to purify the recombinant products rLipL21, rOmpL1/1, rOmpL1/2, rLipL32/1, rLipL32/2, rLipL41/1 and rLipL41/2 expressed by the major genotypes of four leptospiral OEPs of 15 serogroups. SDS-PAGE is applied to examine the expression and purity of the recombinant proteins. Rabbits are intracutaneously immunized with the recombinant proteins to obtain antisera. Microscope agglutination test (MAT) is used to measure the cross inmmunoagglutination titers of antisera. The OMPs of the reference standard strains belonging to 15 serogroups of L. interrogans in China and L. biflexa strain Patoc I are prepared using salt-denature method. By each of the antisera as the first antibody, Western blot assay is established to detect the natural expressions and immunoreactivity of the four OEPs. RESULTS: The outputs of rLipL21, rLipL32/1, rLipL32/2, rLipL41/1l, rLipL41/2, rOmpL1/1 and rOmpL1/2 are 10%, 40%, 35%, 15%, 10%, 30% and 15%, respectively. Each the purified recombinant proteins shows a single fragment after SDS-PAGE. Each the rabbit antisera displays extensive cross immunoreactivity between the products expressed by different genotypes of the same gene and the MAT titers ranging from 1:2-1:128. All the four OEPs can be detectable in the OEPs preparations. However, LipL21 is found to exist only in L. interrrogans. CONCLUSION: The results of this study indicate that all the four OEPs are superficial genus-specific antigens of Leptospira which can be used as the candidate antigens of leptospiral universal vaccine and detection kit. PMID- 17415990 TI - [Study on the relationship between Gln223arg variant in leptin receptor gene and obesity]. AB - OBJECTIVE: To investigate the association between Gln223Arg variant in leptin receptor (LEPR) gene and obesity. METHODS: The subjects were from Yu county, Shanxi province, China. Data on cardiovascular risk factors was collected with questionnaire and their Gln223Arg variants in LEPR gene were genotyped. The relation between Gln223Arg variant in LEPR and obesity was analyzed. RESULTS: The genotype frequencies of G. G, G. A and AA were 0.7679, 0.2171 and 0.015 respectively and the allele frequencies of G and A were 0.8764 and 0.1236. The distribution of genotype was in Hardy-Weinberg equilibrium (P = 0.934). The results of single factor analysis showed that the carriers with the Arg223 alleles had higher weight (63.2 kg vs. 61.9 kg; P= 0.0307) and body mass index (24.4 kg/rn vs. 24.1 kg/m2; P = 0.0898) than those noncarriers of these alleles. Results from multivariate analysis indicated that the carriers with the Arg223 alleles had higher body mass index (24.5 kg/m2 vs. 24.1 kg/m ;P= 0.0396) and prevalence of obesity (OR = 1.30, 95% CI: 0.957-1.767; P= 0.0935). CONCLUSION: These data indicated that Gln223Arg variant in LEPR gene was possibly associated with obesity, respectively. PMID- 17415991 TI - [Study on mannose-binding protein gene polymorphisms and susceptibility to pulmonary tuberculosis]. AB - OBJECTIVE: To explore the association between the genetic polymorphisms of mannose-binding protein (MBP) alleles and susceptibility to pulmonary tuberculosis. METHODS: 125 pulmonary tuberculosis cases and 198 healthy controls were collected. A case-control study was conducted. Three structural gene mutations in exon 1 of MBP gene (codon 52, codon 54 and codon 57) were studied. Polymerase chain reaction with sequence-specific primers (PCR-SSP) was carried out in the polymorphism in MBP alleles. Information on related risk factors of tuberculosis was collected, using a pre-tested questionnaire. Univariate and multivariate logistic analyses were conducted with SPSS software package. RESULTS: The frequencies of mutant heterozygote or homozygote of MBP-52, 54, 57 were 8.0%, 7.2% and 0.4% for cases and 5.3%, 4.3%, 0.5% for controls, respectively. The distribution of mutant genotypes of MBP did not show significant difference between tuberculosis patients and control by Mantel Haenszel chi2 on sex. The univariate analysis demonstrated that body mass index, marital status, vaccinal vestige, bacillus of Calmette-Guerin vaccine immunization, contacted with pulmonary tuberculosis patients, familial traits were the risk factors of pulmonary tuberculosis. After adjusting those related environmental factors in the multivariate logistic analyses, the total MBP (MBP 52, MBP-54 and MBP-57) and MBP-52 heterozygote genotypes were significantly overrepresented in cases, with adjusted OR (95% CI) being 2.182 (1.058-4.499) and 2.574 (1.028-6.446). CONCLUSION: Total MBP and MBP-52 mutant genotypes might be associated with the susceptibility to pulmonary tuberculosis. PMID- 17415992 TI - [A newly discovered the viral zoonoses and related risk factor, prevention and control]. PMID- 17415993 TI - [Countermeasure and progress of microbiological laboratory biosafety management in China]. PMID- 17415995 TI - [Analysis on dependency of daily life and it's relationship with chronic disease among rural elderly population in Anhui Province]. PMID- 17415994 TI - [Interaction between familial cancer history and smoking on the risk of lung cancer in a Chinese population]. PMID- 17415996 TI - [Study on the epidemiological characteristics and preventive measures of rabies in Shanghai from 2001-2005]. PMID- 17415997 TI - [Investigation on a outbreak of hepatitis A]. PMID- 17415998 TI - [Advances in research on biological target volume and conformal intensity modulated radiotherapy]. PMID- 17415999 TI - [Glycogen synthase kinase-3beta (GSK-3beta) promotes proliferation of ovarian cancer cells in vitro]. AB - OBJECTIVE: To investigate the effect of glycogen synthase kinase-3beta (GSK 3beta) on the proliferation of human ovarian cancer cells. METHODS: Two human ovarian cancer cell lines SKOV3 and ES-2 were analysed for the expression of GSK 3beta and phosphorylated GSK-3beta (pGSK-3beta) by Western blot analysis. Cell growth curve analysis done by cell count was used to investigate the effect of GSK-3beta inhibitors on the growth of SKOV3 and ES-2 cells. Four plasmids, namely, GSK-3betaS9A, GID5-6, GID5-6LP and the control vector, were cotransfected respectively with the green fluorescent protein (GFP) into SKOV3 cells by electroporation, and then BrdU incorporation assay was adopted to analyse the role of GSK-3beta activity in the proliferation of ovarian cancer cells. After transfection, G418 was added to the medium to select those stably transfected cells, which were used to investigate the long term effect of GSK-3beta activity change on the proliferation of ovarian cancer cells by colony formation assay. RESULTS: Both SKOV3 and ES-2 cells expressed GSK-3beta, though the expression level of pGSK-3beta was lower in SKOV3 than in ES-2 cells. GSK-3beta inhibitors attenuated the growth of SKOV3 and ES-2 cells. Transfection with GSK-3betaS9A to upregulate the GSK-3beta activity resulted in the increase of BrdU incorporation in SKOV3 cells compared with that in the control vector. On the contrary, transfection with GID5-6 to downregulate GSK-3beta activity decreased the BrdU incorporation in SKOV3 cells, compared with that in GID5-6LP, which is a control vector of GID5-6. Stable transfection with GSK-3betaS9A increased the colony number while stable transfection with GID5-6 decreased the colony number, compared with each control vector. CONCLUSION: GSK-3beta can promote the proliferation of ovarian cancer cells. Inhibition of GSK-3 p may become a potential theraputic PMID- 17416000 TI - [The regulation of EKI-785 to the growth of U937 cell line]. AB - OBJECTIVE: The aim of this study was to explore the expression of erbBs in U937, an acute monocyte leukemia cell line, and their impact on the growth of this cell line. METHODS: Expression of erbBs was detected by RT-PCR and expression of erbB2 at protein level by Western blot. After U937 cells was treated with EKI-785, an irreversible specific inhibitor of erbBs, the growth was assessed by MTT and growth curve, apoptosis was detected by Annexin V-FITC Apoptosis Detection Kit, and signal pathway was detected by Western blot. RESULTS: erbB2-4 were expressed in U937 cell line, but not erbB1. Especially, protein of erbB2 was expressed in this cell line. After treating with EKI-785, the growth of U937 cells was inhibited and early apoptosis was induced. Moreover, the Ras/MAPK and the PI3K/Akt signaling pathways were all blocked. CONCLUSION: erbBs may play key roles in the development of some leukemia. Therefore, erbBs may become new targets of treatment to leukemia, and EKI-785 has a potency of clinic use to leukemia. PMID- 17416001 TI - [The inhibitory effect of recombinant polypeptide CH50 of fibronectin on invasion and angiogenesis of tumors]. AB - OBJECTIVE: To investigate the inhibitory effect of recombinant polypeptide CH50 of fibronectin on invasion and angiogenesis of tumors, and analyze the possible molecular mechanism of the therapeutic effect of polypeptide CH50 on tumors. METHODS: The tumor model was established by inoculation of H22 hepatocarcinoma cells in mice. The tumor gene therapy was performed by in vivo gene transfection with a method based on hydrodynamics to express polypeptide CH50. After treatment, the inhibitory effect on tumor invasion and angiogenesis was observed by histotology with HE staining of tumor tissues. The expresison of MMP-9 mRNA and protein at the edge of tumor tissue was evaluated by RT-PCR and gelatin zymography, respectively. RT-PCR was used to detect the expression of the related genes in H22 cells treated with polypeptide CH50. Cell adhesion assay was used to analyze the influence of polypeptide CH50 on the binding of cells to fibrinogen. RESULTS: (1) Eukaryotic expression plasmid pCH510 was expressed in vivo in a non targeting manner and produced a significant inhibitory effect on tumor growth. The therapy with polypeptide CH50 resulted in pronounced necrosis of tumor cells in pCH510 group, compared with that in control groups at histological level. (2) Polypeptide CH50 could inhibit the growth, invasion and angiogenesis of the tumor, and interfere the formation of new collateral circulation in the tumor. (3) The expression level of MMP-9 protein at the edge of tumor tissue was significantly decreased after treatment, especially the activation of pro-MMP-9 was inhibited significantly, whereas the expression level of MMP-9 mRNA was not influenced. (4) The expression of alphav, 33 and cdc2 mRNAs in H22 cells treated with polypeptide CH50 was down-regulated. (5) Cell adhesion assay manifested that polypeptide CH50 can affect the adhesion ability of H22 cells. CONCLUSION: Polypeptide CH50 can inhibit tumor growth and angiogenesis by suppressing the functions of MMP-9 and integrin alphavbeta3. PMID- 17416002 TI - [Construction of deltaNp63 specific small hairpin RNA expressing plasmid and its role in bladder cancer--a preliminary study]. AB - OBJECTIVE: To construct deltaNp63 specific small hairpin RNA (shRNA) expressing plasmid,to examine its inhibitory effect to the expression of deltaNp63 protein and mRNA in transitional cell carcinoma of the bladder (TCCB) , its effect on TCCB cells cycle and proliferation. METHODS: DeltaNp63 specific oligonucleotides were designed and synthesized. These oligonucleotides were annealed to form double strand DNA fragments and this fragment was cloned into Pgenesil-1 plasmid. The recombinant deltaNp63-shRNA expression construct was confirmed by using Pst I + Sal I double digestion and by sequencing. Fluorescence staining was used to confirm the success of transfection in TCCB cells under the fluorescence microscope. The inhibitory effect of deltaNp63-shRNA construct was examined with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical staining assay. The cell cycle of TCCB cells was assayed by flow cytometry (FCM). The cellular proliferation of TCCB cells was assayed by tetrazolium bromide (MTT) colorimetry. RESULTS: The deltaNp63-shRNA expression plasmid was successfully constructed and transfected into TCCB cells. It can effectively reduce the expression of deltaNp63 protein and mRNA. The reduction rate of deltaNp63 mRNA was 63.0%, and the G0/G1 ratio was increased and S phase was decreased in transfected TCCB cells. The cellular proliferation was also lower in transfected 5637 cells in comparrison with that of non-transfected TCCB cells. CONCLUSION: A deltaNp63-shRNA expression plasmid, constructed from Pgenesil-1 plasmid, can successfully be transfected into TCCB cells and can effectively inhibit the expression of deltaNp63 protein and mRNA. It also can take part in regulation of the cell cycling and inhibit the cellular proliferation of TCCB cells. PMID- 17416003 TI - [Relation between c-erbB1, c-erbB2, MAPK expression and resistance to tamoxifen in breast cancer cells in vitro]. AB - OBJECTIVE: To study the growth regulation pathway and the mechanism of acquired resistance to tamoxifen (TAM) in breast cancer cells. METHODS: TAM was used to induce wild-type MCF-7 human breast cancer cell line and establish a tamoxifen resistant (TAM-R) cell line. RT-PCR, Western blot and immuocytochemical techniques were used to detect and compare mRNA and protein of c-erbB1, cerbB2, c erbB3, c-erbB4 in wild-type MCF-7 and TAM-R MCF-7 cell lines. RESULTS: Compared with wild-type MCF-7 cells, the mRNA of c-erbB1 increased 6 times (P < 0.05) and the protein 3 times higher (P < 0.05), and the mRNA of c-erbB2 increased 3 times (P < 0.05) and the protein 1.5 times higher (P < 0.05) in TAM-R MCF-7 cells. However, comparable levels of c-erbB3 mRNA and protein were expressed in both cell lines. c-erbB4 could not be detected. Under basic conditions, phosphorylated c-erbB1/c-erbB2 and c-erbB1/c-erbB3 heterodimers but not c-erbB2/c-erbB3 receptor heterodimers were detected in TAM-R cells in association with increased level of phosphorylated MAPK. CONCLUSION: Our findings demonstrated that the development of TAM-resistance in MCF-7 cells is related with the autocrine release and action of an c-erbB1-specific ligand inducing preferential c-erbB1/c-erbB2 dimerization and downstream activation of the MAPK pathway. PMID- 17416004 TI - [Assessment of cervical intraepithelial neoplasia (CIN) lesions by DNA image cytometry]. AB - OBJECTIVE: To compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions. METHODS: 87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry. RESULTS: Of the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method. CONCLUSION: The study demonstrated that DNA ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions. PMID- 17416005 TI - [Comparison of treatment planning by carbon ion radiotherapy and by intensity modulated radiotherapy for prostatic adenocarcinoma]. AB - OBJECTIVE: To evaluate the potential benefit of carbon ion radiotherapy (C-ion RT) through comparison with photon intensity-modulated radiotherapy (IMRT) in dose distribution for prostatic adenocarcinoma. METHODS: In randomly selected 5 patients, treatment planning of C-ion RT (4 coplanar beams) and IMRT (7 coplanar fields) were worked out by computer working station. In order to make a meaningful comparison, it was defined that the 95% isodose surface had to cover 100% of the PTV in each plan; all dose was given as normalized dose with the definition of the minimum dose of the PTV being equal to 95% of prescribed dose. Dose-volume histograms (DVHs) of the tumor and organ-at-risks (OARs) were calculated. Volume irradiated more than or equal to some specified doses, conformity index ( CI) , and inhomogeneity coefficient (IC) of each treatment plan was compared, respectively. RESULTS: With C-ion RT, the mean irradiated volumes (in %) of the rectum were significantly smaller than that with IMRT except for 95% dose level, and C-ion RT could provide complete protection to the posterior rectal wall. In addition, C-ion RT could also remarkably reduce the dose to the bladder, femoral heads and non-target normal tissues at each dose level. Dose conformation and homogeneity in the target volume of C-ion RT were better than that in IMRT (mean CI50%, 3.36 vs. 5.04, mean CI95%, 1.20 vs. 1.46, mean IC, 0.03 vs. 0.12). CONCLUSION: Compared with IMRT, C-ion RT can obtain better dose distribution, and may reduce tumor recurrence and radiation-induced complications in prostatic adenocarcinoma. PMID- 17416006 TI - [The effect of amino acid nutritional support on serum tryptophan and melatonin in lung cancer patients receiving chemotherapy]. AB - OBJECTIVE: To investigate the effect of amino acid parenteral nutritional (PN) support on serum tryptophan and melatonin in non-small cell lung cancer (NSCLC) patients receiving chemotherapy. METHODS: Seventy-two patients with inoperable NSCLC were divided into three groups randomly: control group, 250 ml/d amino acids PN therapy group and 500 ml/d amino acids PN therapy group. The same NP (cisplatin + vinorelbine) chemotherapy was carried out in all the three groups. During three sessions of chemotherapy,amino acids PN therapy was given to the amino acids PN therapy groups. Serum tryptophan and melatonin concentration changes were assessed before and after chemotherapy. RESULTS: After chemotherapy the concentration of MT and Try were much lower than that before chemotherapy in the three group patients (P < 0.05). But the concentration of MT and Try in the PN group patients was higher than that in control group patients. The concentration of MT and Try in the 500 ml/d amino acid parenteral nutritional support group patients were significantly higher than that in the 250 ml/d group patients, the difference was significant (P < 0.05). CONCLUSION: Amino acid parenteral nutritional support is beneficial to improve the lower concentration of serum MT and Try in NSCLC patients receiving chemotherapy, and a more significant effect can be achieved by the 500 ml/d amino acid parenteral nutritional support treatment. PMID- 17416007 TI - [Ginsenosides and dexamethasone in managing the liver injury and renal function after transcatheter arterial chemoembolization for hepatic carcinoma patient]. AB - OBJECTIVE: To observe the protective effect of ginsenosides (GS) or low dose of glucocorticoid dexamethasone (Dex) alone or combined in managing the liver injury and renal function after transcatheter arterial chemoembolization (TACE). METHODS: 120 patients with primary liver carcinoma were randomly divided into four groups (A, B, C, D) with 30 patients in each. Group A was treated with placebo; group B with Dex; group C with GS and group D with Dex plus GS. The changes in liver and renal function after TACE were observe according to the WHO criteria for side effects of anti-cancer drug. RESULTS: Compared with group A, Dex combined with GS was able to reduce the level of TB, ALT/AST, BUN and Child grade, which significantly protected the liver and kidney (P < 0. 05). However, Dex or GS alone could also improve some parameters of liver and renal function after TACE (P < 0.05). CONCLUSION: Dex combined with GS is effective in managing the liver injury and renal function after transcatheter arterial PMID- 17416008 TI - [Observation and clinical significance of adjuvant chemotherapy-induced amenorrhea in premenopausal breast cancer patients]. AB - OBJECTIVE: A retrospective analysis of 160 pre-menopausal breast cancer patients was carried out to elucidate the the menstrual outcome in those cases who had undergone adjuvant chemotherapy after surgery, and evaluate the relationship between chemotherapy-induced amenorrhea (CIA) and recurrence of the disease. METHODS: 160 pre-menopausal breast cancer patients were collected, 62/159 (39.0%) of them were node positive, 91/158 (57.6%) were ER positive, and 95/155 (61.3%) were PR positive. 111 cases had infiltrative ductal carcinoma, 26 cases had infiltrative lobular carcinoma, and 22 cases with others. In 152 cases data were collected by face-to-face interview and 8 cases by phone conversation. Types and cycles of chemotherapy regimen as well as menstrual abnormalities were recorded before, during, and after chemotherapy completion. Follow up duration was 12-72 months after chemotherapy completion for all patients. RESULTS: 107 (66.9%) developed CIA, 24 cases returned to normal menses (22.4%), 83 cases continued CIA during more than 12-month follow up (77.6%). The rate of CIA increased with age (P < 0.01). During the follow up, disease free survival (DFS) rate was 85.9% in CIA group and 79.2% in non-CIA group, with no statistically significant difference. But in hormonal receptor positive patients, DFS was 80.0% in non-CIA and 90.1% in CIA, respectively (P = 0.04), showing a significant difference. Because of the small number of died cases, no analysis of the overall outcome was carried out. CONCLUSION: Adjuvant chemotherapy causes ovarian function suppression, and may further leading to amenorrhoea. Women who experienced amenorrhoea after chemotherapy had a significantly better disease-free survival (DFS) rate showed by univariate analysis than women who continued normal menstruation. Chemotherapy is insufficient therapy for very young patients who are in high risk with hormone responsive disease, particularly when chemotherapy fails to induce amenorrhea. Further research is needed to evaluate interventional chemotherapy to improve the quality of life in women with early stage breast cancer who experienced ovarian toxicity. The post-chemotherapy menstruation status is a clinically valuable, objective and salient marker for sufficient endocrine effect of chemotherapy in ER/PR-positive premenopausal patients. PMID- 17416009 TI - [Clinicopathological characteristics and prognosis of remnant stomach cancer- report of 45 cases]. AB - OBJECTIVE: To investigate the clinicopathological characteristics and prognostic factors of remnant stomach cancer. METHODS: The clinicopathological and prognosis data of 45 patients with remnant stomach cancer were retrospectively analyzed. RESULTS: The remnant stomach cancer are likely to develop in males with a ratio of male to female: 44:1. Their initial operation modes of these patients were Billroth II subtotal gastrectomy in 40 patients, Billroth I subtotal gastrectomy in 4 and proximal subtotal gastrectomy in 1. The interval from the initial operation to the diagnosis of remnant stomach cancer was 5 to 42 years with an average of 23 years. Of these 45 patients, 28 had lesion at anastomotic site, 9 in the gastric cardia and 8 in other locations; 19 had radical resection, 16 palliative resection and 10 exploration alone except one who had an anastomosis of remnant stomach with the jejunum. The histology types included: 1 un differentiated adenocarcinoma, 36 poorly-differentiated adenocarcinoma, 7 moderately-differentiated adenocarcinoma and 1 well-differentiated adenocarcinoma. The 1-, 3-, 5-year survival rates of patients with radical resection were significantly better than those treated with palliative resection, which was 100% vs. 62.5%, 78.8% vs. 25%, 47.2% vs. 0, respectively (P < 0.05). All ten patients without resection died within 2 years with an average survival time of 12 months. The 5-year survival rate of stage I, II, III and IV was 100%, 75%, 17.8% and 0, respectively (P < 0.05). CONCLUSION: Remnant stomach cancer prevalently occurs in the male usually 10 years after Birroth II gastrectomy. The lesions is mainly located at anastomotic site. Poorly-differentiated adenocarcinoma is found to be the prevalent histological type of advanced remnant stomach cancer. The prognosis of remnant stomach cancer is correlated with pTNM stage and whether having been treated with complete resection or not. Patients with early remnant stomach cancer may survive for a long time if radical resection can be done. PMID- 17416010 TI - [Expression of activated ERK1/2 and cyclin D1 in oral squamous cell carcinoma]. PMID- 17416011 TI - [Significance of multidisplinary surgery in chest wall resection and reconstruction for selected patients with breast cancer]. AB - OBJECTIVE: To evaluate the clinical significance and survival benefits of chest wall resection and reconstruction (CWRR) by multidisciplinary surgery for selected patients with locally advanced or recurrent breast cancer in order to address the importance of collaboration between ablative (breast surgery or/and thoracic surgery) and reconstructive teams during CWRR. METHODS: The data of 44 patients who underwent multidisciplinary CWRR at The University of Texas M. D. Anderson Cancer Center between March 2001 and June 2004 were retrospectively analyzed, which included the CWRR techniques used, patient characteristics and treatment outcomes. Survival of patients with primary, recurrent or metastatic disease, and that of those with curative or palliative resection were also compared. RESULTS: All patients were female aged 34-83 years with primary (n=19), recurrent (n=15) or metastatic breast cancer (n=10). The surgery modes included curative resection(n=36) and palliative (n=8) with a mean defect size of skin: 218.4 cm2; of bony chest wall: 113.9 cm2 (n=15). Immediate reconstruction (n=43) with prosthesis (n=10) or without (n=34) for most of these patients. All of them required soft tissue coverage with pedicled flap (n=37) or free flap (n=13). The average hospital and ICU stay was 6.6 days and 3.4 days (n=8), respectively. The morbidity was acceptable and no 30-day mortality happened. Neither was there difference in median survival (44.7 m vs. 36.0 m, P = 0.752) nor in 1-, 2-, and 3 year survival rates between primary breast cancer (78.4%, 78.4%, 39.2%) and recurrent one (70.9%, 70.9%, 70.9%, P > 0.05). However, both median survival (16.0 m) and the 1-, 2-, and 3-year survival rates (30.0%, 15.0%, 0) in patients with metastasis were much poorer than that in those with primary breast cancer (P = 0.003) or recurrence(P =0. 018). The survival of patients underwent curative resection (36.0 m, 71.5%, 65.7%, 65.7%) were much longer than those with palliative resection (15.1 m, 35.1%, 23.4%, 7.8%, P = 0.018). CONCLUSION: With full control of systemic and local disease by up-front multidisciplinary strategy, chest wall resection and reconstruction could improve long-term survival if curative resection achieved or the quality of life if palliative resection done for breast cancer patients with tumor invading the chest wall or local recurrence. Ablative and reconstructive surgeons should be included in surgery team in order to guarantee the possibility of extensive resection and effective reconstruction in a single stage with high safety, good survival and minimal morbidity. PMID- 17416012 TI - [Value of cardiopulmonary bypass for locally advanced lung cancer involving the left atrium]. AB - OBJECTIVE: To review the experience and assess the value of cardiopulmonary bypass (CPB) in the treatment of locally advanced lung cancer involving the left atrium. METHODS: From Jan. 1999 to Dec. 2002, lobectomy or pneumonectomy combined with partial resection of the left atrium were carried out in 52 such patients, which included 13 with assistance of CPB and 39 without. RESULTS: There was one postoperative death in each of the CPB and non-CPB groups due to brbonchopleural fistula and pulmonary infection. Six patients in CPB and 14 in non-CPB groups developed postoperative cardiac complication including arrhythmia, pneumonia and heart failure. The 1-, 3-year survival rate of CPB and non-CPB groups was 69.3%, 66.7% and 38.5%, 38.5%, respectively. CONCLUSION: Combined resection of locally advanced lung cancer with partially involved left atrium through cardiopulmonary bypass was safe and effective, and may not increase the postoperative complication and risk. PMID- 17416013 TI - [Primary germ cell tumor in the mediastinum-report of 47 cases]. AB - OBJECTIVE: To investigate the clinical characterstics, effective treatment and prognosis in patients with primary germ cell tumors in the mediastinum. METHODS: The data of 47 such patients treated from 1967 to 2005 were retrospectively reviewed. RESULTS: Of these 47 patients, 41 were male and 6 female with a median age of 26 years; 8 (17.0%) had seminoma, and 39 (83.0%) non-seminoma. The overall 1-, 3-, 5-year survival rate was 63.4%, 37.5% and 34.8% with a median survival time of 16 months; which was 100%, 83.3% and 83.3% for seminoma and 56.4%, 30% and 27.3% for nonseminoma, respectively. The pathologic type was found to be the only independent prognostic factor (P = 0.045). CONCLUSION: Primary mediastinal serminoma is sensitive to radiotherapy or chemotherapy with a good prognosis, but the prognosis of primary mediastinal non-seminoma is poor. Cisplatin-based chemotherapy still plays a key role in the treatment of primary mediastinal non seminoma as the survival of those has been improved. PMID- 17416014 TI - [Assessment of neoadjuvant chemotherapy in breast cancer patients]. AB - OBJECTIVE: To assess the response of neoadjuvant chemotherapy and its influencing factors in the breast cancer patients. METHODS: 171 patients with stage II or operable stage III breast cancers were treated with neoadjuvant chemotherapy before surgery between January 2004 and May 2005. Of these, 160 received and completed > or =3 cycles of neoadjuvant chemotherapy, 11 received only 2 cycles. The regimens of neoadjuvant chemotherapy were: CEF (CTX, Epirubicin, 5-Fu); NE (Navelbine, Epirubicin); TEC (Taxotere, Epirubicin, CTX). Response of neoadjuvant chemotherapy was evaluated in all patients by palpation, ultrasonography and pathological methods. RESULTS: Complete response rate and clinical objective response rate determined by clinical palpation (cCR, cOR), ultrasonography (sCR, sOR) and pathology (pCR) was 18.7% and 88.3%; 4.1% and 74.9%; 15.2%, respectively. The correspondence rate of the pCR with cCR and sCR was 43.8% and 42.9%, respectively. It was showed by univariate analysis that patient whose tumor was < or =3 cm in diameter, or ER negative or grade 3 were more likely to achieve a pCR than those whose tumor was >3 cm, or ER positive or grade 1. Logistic regression analysis showed that only tumor size was the significant predictive factor for response to neoadjuvant chemotherapy in patients with primary breast cancer. CONCLUSION: Patient with small, or ER negative or grade 3 tumor may have better pathological response to neoadjuvant chemotherapy, particularly, the tumor size is more predictive of pCR. Palpation or ultrasonography may have a tendency either to under- or to overestimate pCR. Breast neoplasms/drug therapy; PMID- 17416015 TI - [Impact of extracapsular lymph node spread in the ipsilateral neck on contralateral neck metastasis and prognosis of laryngeal cancer]. AB - OBJECTIVE: To evaluate the impact of extracapsular lymph node spread (ECS) in the ipsilateral neck on the contralateral neck metastasis and prognosis of laryngeal cancer. METHODS: The data of 184 laryngeal cancer patients who underwent laryngectomy and simultaneous radical or modified radical neck lymph node dissection between Jan. 1994 and Dec. 1997 were retrospectively analyzed. Of these 184 patients, 144 underwent unilateral neck lymph node dissection and 40 bilateral; 159 had supraglottic lesion and 25 transglottic. All had squamous cell carcinoma. The clinical T stage was T1 in 3, T2 63, T3 77, T4 41; N stage: NO in 123, N1 38, N2a 5, N2b 11, N2c 7. Transparent lymph node detection and continuous sectioning method were applied to all dissected neck lymph nodes. Statistical analysis was carried out using SPSS software package ( version 11.5). Survival curves were calculated through the Kaplan-Meier model. Impact of extracapsular lymph node spread in the ipsilateral neck on prognosis was assessed using the Log rank test. RESULTS: Of these 184 patients, neck lymph node metastasis was pathologically proven in 80, 26 had ECS in the ipsilateral neck with a ECS rate of 32.5% (26/80). The ECS incidence was positively correlated with advanced pathological N stage and metastatic lymph nodes (P < 0.01). The incidence of the contralateral neck metastasis and ipsilateral neck recurrence with ECS were higher than those without ECS, which was 46.2% versus 24.1%, and 34.6% versus 7.4%, respectively (P < 0.05). The 3- and 5-year survival rates of patients with ECS were significantly lower than those of patients without ECS, which was 53.9% versus 70.4%, and 23.1% versus 57.4%, respectively (P = 0.0125). CONCLUSION: Extracapsular lymph node spread is found to be an important prognostic factor in the laryngeal cancer. Bilateral neck dissection may be mandatory due to patients with ECS have a higher incidence of contralateral neck metastasis. The capsule of metastatic lymph nodes should be pathologically checked and reported in order to determine the extra-capsular spread status. PMID- 17416016 TI - [The application of multi-slice CT dynamic enhancement scan in the diagnosis and treatment of colonic lymphomas]. AB - OBJECTIVE: To discuss the value of multi-slice CT dynamic enhancement scan in the diagnosis and treatment of colonic lymphomas. METHODS: 16 patients with colonic lymphomas underwent multi-slice CT dynamic enhancement scans, images of axial and reconstructive images of VR, MPR and CTVE were analyzed, patients were respectively diagnosed. RESULTS: Appearances of primary colorectal lymphomas were categorized into focal and diffuse lesions. Focal and diffuse lesions were 6 and 10 patients, respectively. The accuracy rate of diagnosis was 87.5%. CONCLUSION: MSCT dynamic scan has distinctive superiority in diagnosis and treatment of colonic lymphomas. PMID- 17416017 TI - [Perspectives on clinical studies of cancers of the esophagus and gastric cardia from the point of view of high incidence field research]. PMID- 17416018 TI - An invitation to do the write thing. PMID- 17416019 TI - The HPV vaccine: a major public health breakthrough. PMID- 17416020 TI - Factors to consider before referring patients for major cancer surgery. PMID- 17416021 TI - The nonoperative management of shoulder instability. PMID- 17416022 TI - An unusual rash in an elderly man. PMID- 17416023 TI - Type 2 diabetes in children and adolescents: screening, diagnosis, and management. PMID- 17416024 TI - Circumferential electrical contact burn of a finger. PMID- 17416025 TI - Pelvic pain, vaginal spotting, and a right adnexal mass. PMID- 17416026 TI - A mysterious patch of white hair on a young boy's scalp. PMID- 17416027 TI - Case of the month. Septic arthritis with concomitant gas-producing myonecrosis in the periarticular soft tissues of his knees. PMID- 17416028 TI - [Nutricion Hospitalaria included in Science Citation Index Expanded. Impact factor for 2008]. PMID- 17416029 TI - [The MEI network unites Latin America around malnutrition]. PMID- 17416030 TI - Effects of a very low birth weight newborn on family: literature review. AB - OBJECTIVE: The present study is a literature review to identify the effects of a very low birth weight newborn on family. This is an important instrument to clarify epidemiological issues and to suggest the directions for health policy efforts. METHOD: A three-step review was carried out using databases of journals indexed for Medline/Lilacs/ Scielo/Cochrane published between 1966 and 2005 using specific criteria of inclusion. The first step selected 12 articles from 2889 when searching for the keyword "very low birth weight infant"; the second step used the crossing of keyword "premature infant" with other pertinent keywords and terms resulting in 191 articles generating 7 more articles matching the criteria of inclusion. The third step was to analyze the references of articles in steps 1 and 2 (12 + 7 = 19), selecting 3 additional ones totalizing 22 selected articles. RESULT: Evidences in literature state that the families of very low birth weight newborns suffer potential negative effects on their operational dynamics, which is associated to the clinical seriousness, the age and the neuropsycomotor development of such children. It seems that the mother is the most affected member due to the situation imposed to the family, and the one who needs psychosocial support more frequently. CONCLUSION: The number of existing studies is still insufficient to clarify whether the effects on the family considering all their aspects are preponderantly positive or negative. PMID- 17416031 TI - [Animal models for the study of systemic inflammatory response and parenteral nutrition]. AB - The systemic inflammatory response syndrome (SRIS) seems to be due to the activation of the toll-like receptors, specific of the inflammatory response cells, through concrete cytosolic signals which lead to a cascade of reactions acting cytokins, growing factors and others inflammatory mediators. This kind of work revewes and discusses several classifications of animals models to study the SRIS, and propose to divide these models according to concrete goals, which can be the following ones: (1) To study innate and adaptative receptors of regulatory gens in the SRIS. (2) To study signals receptors (cytokines and growing factors). (3) To study the answer to signals. (4) To study treatments through specifics antinflammatory blockage. (5) Specific models of sepsis. (6) Others inducing models of SRIS. (7) Others therapeutical models. -Antinflammatories. Antiacoagulans: Coagulations inhibition in human assays. Phase II Anticoagulans: Antitrombine III, PCA and TFPI. -Antibiotics. -Replacing Volume Treatments. Surgical Treatments. As to the animals models to study Parenteral Nutrition, we could make the next classifications and sum it up: (1) Animal models to study the parenteral via of administration. (2) Models to study viability, absorption and local tolerance of the administration via. (3) Study models for complications. (4) Animal models to study pharmacodynamic, metabolization and to investigate the tolerance of new molecules or substrates. PMID- 17416032 TI - [Animal models in glucose intolerance and type-2 diabetes]. AB - There stilla are many unknown facts about the pathogenic mechanisms of such a prevalente disease nowadays as i type-2 diabetes mellitus (DM2). The advances in diabetes prevention and management will greatly depend on the understanding of these mechanisms; therefore, it will be essential to keep on using animal models on which carrying out experiments that would be urethical in humans. DM2 represents a heterogeneous group of diseases characterized by an increase in insulin resistance at periphera tissues and a deterioration in insulin secretion by pancreatic beta-cells, both abnormalities being highly interweaved. The mentioned heterogeneity of DM2 is also reflected by the high diversity of useful animal models for its study. The main DM2 models are reviewed, classifying them by their mechanism of action in spontaneous or induced. Also, two categories in each one of them are distinguiseed: analogous models, which try to imitate the human disease, and intrinsic models, with which we pretend to answer specific questions about the disease. The decision about which model to case for a particular experiment usually is multifactorial. Ideally, the experiments should be carried out in several different models, taking into account that none of them completely reflects the complexity of human DM2 and that precautions should be taken when trying to extrapolate the findings to the clinical practice. PMID- 17416033 TI - [Experimental models of cardiovascular disease]. AB - The present work describes clinically useful experimental models for the study of cardiovascular disease and emphasites the models used to determine the pathophysiologic mechanisms of atherosclerosis, as well as to evaluate the effects of nutritional and pharmacological products on the development of this complex inflammatory process present in many cardiovascular diseases. Animal models in which ahterosclerosis may be induced by dietary changes are reviewed, as well as those in which modification in one or more genes (knock-out and knock in animals), or the incorporation of foreign genes from other species lead to early cardiovascular disease. On the other hand, some of the cell lines most frequently used in studying molecular mechanisms of atherosclerosis and assessment of substances with nutritional or pharmacological interest are considered. PMID- 17416034 TI - [In vivo experimental models of inflammatory bowel disease and colorectal cancer]. AB - There is no ideal animal model for the study of human pathologies such as IBD or CRC. For the last decade, genetically engineered animal models have been incorporated to models induced by exogenous agents. Experimental models of IBD reproduce heterogeneous intestinal inflammatory conditions, as it occurs in the human being, whereas CRC models imitate those mutations found in man although with different phenotypic repercussions. In both cases, these experimental systems are influenced by genetics and the environment. Today, animal models represent valued, complex, and complete bioassays for the study of new therapeutic strategies in IBD (IL-10, anti-IL-12, probiotics) and protective agents in CRC (n-3 fatty acids, NSAIDS, and COX-2 inhibitors). PMID- 17416035 TI - [Experimental models on hemorragic shock]. AB - This review addresses the pathophysiology and treatment of hemorrhagic shock, a condition produced by rapid and significant loss of intravascular volume. Hemorrhagic shock may lead sequentially to hemodynamic instability, decreases in oxygen delivery, decreased tissue perfusion and cellular hypoxia. Multiple organ failure, a systemic inflammatory process that leads to dysfunction of different vital organs, is a frequent complication after hemorrhagic shock and accounts for a high incidence of mortality. The pathogenesis of organ injury secondary to hypovolemic insults is still not completely understood, but both experimental studies and clinical observations indicate that macrophages are activated by translocated endotoxin-bacteria and ischemia/reperfusion. Activated Kupffer cells release pathologically active substances such as inflammatory cytokines, reactive oxygen species, and nitric oxide, all of which may participate in the mechanisms of hemorrhagic shock. Moreover, increased free radical production during hemorrhagic shock and resuscitation gives place to an increase in oxidative stress that would contribute to the organ damage. In the last few years, a number of experiments have been performed in an attempt to understand the pathophysiology and treatment of hemorrhagic shock. Different studies have shown positive effects on hemorrhagic shock treatment by antioxidant, amino acid, and lipid administration. PMID- 17416036 TI - [Animal models of fulminant hepatic failure]. AB - Fulminant hepatic failure (FHF) is a very serious clinical sindrome that, in spite of the important therapeutical advances that have taken place in the last years by means of bioartifical hepatic support devices and hepatic transplantation, is still associated to a high mortality. Knowledge and treatment of the FHF have been limited by the lack of satisfactory animal models. Among the attempts to develop a suitable model are surgical models, such as hepatectomy and total and/or partial devascularization, or the use of chemical substances with hepatic toxicity, such as acetaminophen, azoximethane, galactosamine or thioacetamide, among others. However, most of these models do not adequatly reflect the pattern of the human disease and all of them present important limitations. Although viral hepatitis is one of the most frequent causes of FHF, the use of viral agents to develop animal models has been little and unfortunate. Our group has recently developed a viral animal model of FHF by means of the inoculation of rabbits with the virus of the rabbit hemorrhagic disease. This model displays biochemical, and histological characteristics, and clinical signs that ressemble those in human FHF. In the present article, the most widely used animal models of FHF, together with their main advantages and disadvantages, are presented. PMID- 17416037 TI - [IV Debate Forum of the SENPE: questions on nutrition research]. AB - The IV Debate Forum of the SENPE values from a multidisciplinary and multi professional perspective the questions on clinical research in nutrition in Spain, focusing on enteral nutrition due to the lack of legislation on this issue. The concluding remarks point out the SENPE commitment with promoting quality, education and facilitation of research, greater help to emergent groups, looking for financial support, and timely information on the several summons from public systems and reference centers PMID- 17416038 TI - [II BAXTER-SENPE workshop: peripheral parenteral nutrition]. AB - During the II BAXTER-SENPE workshop, a multidisciplinary nutrition expert committee discusses on indications, advantages and drawbacks of Peripheral Parenteral Nutrition (PPN), being defined as PN compounded by the three essential elements, vitamins, and minerals. Its composition implies a CH: lipids ratio of 60:40, a limited lipid daily intake (1.3 g/kg of body weight/day) and no more than 30% of total calories), and a nitrogen load not higher than 10 g of nitrogen in a maximum volume of 2500 mL. PPN can be administered by the peripheral route since its osmolarity is lower than 800 mOsm/L and pH is 6.0-7.4; in this way, possible adverse affects of central venous accesses are prevented. Currently, PPN by represent up to 50% of all PNs prescribed at a hospital Studies on efficacy, safety and pharmacoeconomic show that PPN is very useful therapeutic resource in certain clinical situations, both medical and surgical, to prevent metabolic nutritional worsening of the patient, thus having an impact on significant reduction of complications, the number of interventions, and hospital stay. PMID- 17416039 TI - [Nutritional support in patients with open abdomen]. AB - OBJECTIVE: We have designed a protocol to evaluate the tolerance, effectiveness and the nutritional support in post-surgical patients with an open abdomen. PATIENTS AND METHODS: We have made a prospective descriptive study of patients submitted to surgery and left with an open abdomen with a Bogoti bag, and have received nutritional support. The patients who have stayed for less than 4 days with the open abdomen where excluded. A group received total parenteral nutrition (TPN) with mixtures all in one; another group received enteral nutrition (EN) through a needle catheter jejunostomy (NCJ) and a third group received mixed nutritional support. To evaluate the tolerance to EN, we have included those patients receiving this type of nutrition for at least 4 consecutive days without having diarrhea or pain. A jejunostomy catheter was placed in all patients by need o puncturing during the surgical act. SETTING: The Surgical Department I of Angel Larralde University Hospital, I.V.S.S. Valencia-Venezuela, during the period from May 2002 to May 2005. RESULTS: 24 patients entered the study, 46% recived mixed nutritional support (Enteral and Parenteral), 33% exclusively TPN, and 31% exclusively EN. 75% have evolved favorably, discharged to their home in good general condition; 25% died from multiple organ failure due to severe sepsis. About nutritional support, 66% of the patients did not present complications; 21% of those receiving TPN presented hyperglycemia; and 13% of those receiving EN presented diarrhea. About EN effectiveness, 69% of the patients have reached 80% of the estimated caloric objective within days 4-5 from the beginning of formula administration. CONCLUSIONS: The integral care of the patients with an open abdomen, added to a nutritional support regimen tailored to each patient's condition can help decreasing the hypermetabolic response, as well as moridity and mortality. PMID- 17416040 TI - [Does beer have an impact on weight gain? Effects of moderate beer consumption on body composition]. AB - BACKGROUND: The effects of alcohol consumption on body weight have been evaluated in a large number of studies suggesting to be inversely as well as positively related to body weight and body fat. OBJECTIVE: This study examined the relationships between moderate beer consumption and anthropometrical parameters in Spanish healthy adults. METHOD: After a 30 day alcohol abstemious period, 58 healthy volunteers were submitted to a daily moderate consumption of beer during the following 30 days. Weight, height, skinfolds and circumferences were measured at three points: (a) basal; (b) abstemious and (c) after moderate consumption of beer. RESULTS: Biceps skinfold (mm) increased (p < 0.05) in men after moderate beer consumption in regard to abstinence period (5.74 +/- 1.70 vs. 6.23 +/- 1.74). No significant differences were observed in the rest of anthropometrical parameters studied in both women and men along the study intervention. CONCLUSION: Moderate beer consumption during one month did not mostly change weight and weight-related parameters in healthy adults. PMID- 17416041 TI - Recommendations from SPNS/GEAM/SENBA/SENPE/AEDN/SEDCA/GESIDA on nutrition in the HIV-infected patient. AB - OBJECTIVE: to make recommendations on the approach to nutritional problems (malnutrition, cachexia, micronutrient deficiency, obesity, lipodystrophy) affecting HIV-infected patients. METHODS: these recommendations have been agreed upon by a group of expertes in the nutrition and care of HIV-infected patients, on behalf of the different groups involved in drafting them. Therefore, the latest advances in pathophysiology, epidemiology, and clinical care presented in studies published in medical journals or at scientific meetings were evaluated. RESULTS: there is no single method of evaluating nutrition, and diferent techniques--CT, MRI, and DXA--must be combined. The energy requirements of symptomatic patients increase by 20-30%. There is no evidence to support the increase in protein or fat intake. Micronutrient supplementation in only necessary in special circumstances (vitamin A in children and pregnant woman). Aerobic and resistance excercise is beneficial both for cardiovascular health and for improving lean mass and muscular strength. It is important to follow the rules of food safety at every stage in the chain. Therapeutic intervention in anorexia and cachexia must be tailored, by combining nutritional and pharmacological support (appetite stimulants, anabolic steroids, and, in some cases, testosterone). Artificial nutrition (oral supplementation, enteral or parenteral nutrition) is safe and efficacious, and improves nutritional status and response to therapy. In children, nutritional recommendations must be made early, and are a necessary component of therapy. CONCLUSION: appropriate nutritional evaluation and relevant therapeutic action are an essential part of the care of HIV-infected patients. PMID- 17416042 TI - Recovery from experimental malnutrition with soymilk: immunological and genetic aspects. AB - Experimental malnutrition models have been useful to study the effects of malnutrition at early ages. Substantial evidence exists that malnutrition in critical stages of development could result in chromosomal damages. The effect of nutritional rehabilitation with soymilk as a complement of a restricted diet, on plasma and muscle proteins, chromosomal integrity, and unspecific and mucosa immune responses, was studied. Adult male and female Wistar rats (5 weeks old) were assigned to different nutritional conditions: (a) 14 days on protein restricted diet (corn flour and water), followed by 14 days in which water was replaced by soymilk, as nutritional rehabilitation; (b) the same conditions above but periods of 28 days of a protein restricted diet, and 28 days of nutritional rehabilitation and (c) age-matched malnourished (protein restricted diet without nutritional rehabilitation) and normally nourished controls. After both nutritional rehabilitation periods, the weights reached were significantly higher (p < 0.001) than the malnourished control values, but lower than the normal control ones. Plasma protein concentrations were similar in all groups. Muscle proteins that were diminished during the restricted diet, reached normal control values after both rehabilitation periods. The protein restricted diet, produced numeric and structural chromosomal abnormalities. Nutritional rehabilitation was only partially able to revert these abnormalities. The phagocytic activity and gut mucosa IgA-secreting cells were significantly reduced (p < 0.001) during the restricted diet; both nutritional rehabilitation periods induced a significant increase of both, phagocytic activity and IgA secreting cells. These values were similar to controls. Our results show that the supplementation of a protein restricted diet with soymilk improved tissue protein content, as well as unspecific and gut mucosa immune responses, even though it was not able to reinstate fully normal body weight and a normal chromosome karyotype. PMID- 17416043 TI - [Urolithiasis]. AB - In 2006, 70 to 80% of calculi are treated with extracorporeal lithotripsy, 7% with percutaneous nephrolithotomy, 5 to 15% with ureteroscopy and less than 1% with laparoscopic surgery or open procedure. Twenty years after its introduction, extracorporeal lithotripsy is still predominantly used. Today though, its indications are better defined. At the same time, considerable advances have been made in the field of intracorporeal lithotripsy, which have contributed to the development-of ureteroscopy and percutaneous nephrolithotomy. Nowadays, urologists have to master all surgical techniques to best adapt their therapeutic decisions. Finally, they should always keep in mind that treatment for urolithiasis is not limited to surgery. A medical management is essential to prevent recurrences; it concerns all patients with lithiasis, from the very first event, and it is based on a systematic screening for risk factors. This first intention etiological study is carried out in an ambulatory setting, it is easy and cost-effective. It essentially consists in a morphological and constitutional analysis of the calculus by infrared spectrophotometry. Once risk factors for nephrolithiasis have been identified, measures to readjust patient diet and a medical treatment, when necessary, can be implemented. PMID- 17416044 TI - [Sneddon syndrome]. PMID- 17416045 TI - [Williams-Beuren syndrome]. AB - Williams-Beuren syndrome (MIM 194050) is a rare genetic disease (incidence: 1/20 000) which is rather easy to identify during childhood. This developmental disorder associates cardiac defect (supra valvular aortic stenosis most often) in 75% of cases, suggestive face dysmorphism and specific cognitive and behavioural profile. Cognitive profile consists of visuo-spatial bearing deficits contrasting with a relatively well-preserved language. Affected children present with a hyper social behaviour, especially toward strangers; they have a distinctive sensitivity to noise and music. Arterial tension and renal functions have to be looked after during the whole life. Medical and educative management must be done on a multidisciplinary way combining pediatric, cardio-pediatric, orthodontic and psychological approach. Adult patients are generally not completely self sufficient. Williams-Beuren syndrome is due to a chromosomal microdeletion in the q11.23 region of one chromosome 7. It can not be seen on a conventional karyotype and is detected by a FISH (fluorescent in situ hybridization) analyse. This microdeletion leads to the suppression of many genes, mainly the elastin gene. PMID- 17416046 TI - [Iron overload. Diagnosing and treating prior to complications]. PMID- 17416047 TI - [Iron metabolism]. AB - Iron is required for proper cell life functioning. Iron metabolism dysfunction leads in humans to deleterious situations. Maintenance of correct iron plasmatic bioavailability is crucial to permit adequate iron addressing to the different cell types. This implicates especially macrophages and enterocytes, ensuring the import of iron into plasma, as well as systemic signals, including hepcidin a peptide mainly produced by hepatocytes and secreted in plasma, which modulates iron leakage from these cells into plasma. The control of intracellular iron content is under the dependence of the IRE/IRP system which modulates cellular iron ingress and storage. The description of new iron metabolism genes, including hepcidin, paves the road for novel diagnostic tools and therapeutic strategies in the field of diseases associated with iron metabolism abnormalities. PMID- 17416048 TI - [Diagnosis of iron overload]. AB - Hepatic iron overload conditions can be classified as genetic, mainly related to HFE haemochromatosis, and secondary, mainly associated with haematological and hepatic disorders and with metabolic syndrome. The strict affirmation of iron excess relies upon liver biopsy, MRI or calculation of the amount of iron removed through phlebotomies. Determination of its cause relies upon the assessment of transferrin saturation which, when increased, suggests the diagnosis of either haemochromatosis--implying HFE testing--or overload secondary to dysmyelopoiesis or to end-stage cirrhosis, and, when normal, suggests the diagnosis of dysmetabolic iron overload syndrome. PMID- 17416049 TI - [Practical management of HFE hemochromatosis]. AB - HFE-linked, or type 1, hemochromatosis is by far, in the causasians, the most frequent form of chronic iron overload of genetic origin. Its practical management has been recently defined by the French Agency "Haute Autorite de sante". It rests upon a new classification of phenotypic expression. This staging comprises 5 grades according to the combined results of plasma transferrin saturation, ferritinemia and clinical data. The extension of the initial work-up as well as the follow-up modalities is adapted to the staging. A liver biopsy remains indicated only in case of suspected cirrhosis. Venesection therapy is the reference treatment. It is indicated when grade is > or = 2 (i.e. when both transferrin saturation and ferritinemia are increased). The phlebotomy volume should be adapted to the patient's weight. The goal of depletion treatment is to obtain and maintain ferritinemia < or = 50 microg/L. Venesections, especially for maintenance therapy, can be performed at home, which requires a close coordination between the various medical and paramedical partners. Family screening, based upon HFE test, transferrin saturation and serum ferritin, is highly recommended. PMID- 17416050 TI - [Iron overload and chronic liver diseases]. AB - Several liver diseases can be associated with liver iron overload: alcohol liver disease, non alcoholic fatty liver disease, chronic viral hepatitis and porphyria cutanea tarda. These diseases are frequently associated with abnormal iron tests but iron overload is infrequent. Hyperferritinemia in these patients can be explained by cellular necrosis, synthesis increase, inflammation and iron overload. Research of C282Y mutation should be performed in patients who stopped alcohol for at least 1 or 2 weeks and transferrin saturation > 45%. Several studies are conducted to explore the mechanisms of iron overload and the role of iron in hepatic fibrogenesis and carcinogenesis. These studies will help to better define the role of venesection and iron chelators in patients with chronic liver diseases. PMID- 17416051 TI - [Iron overload and arthropathy]. AB - Arthropathy is a major and distinctive manifestation of hemochromatosis, resembling degenerative joint disease with involvement of unusual articular sites, almost identical to the arthropathy in calcium pyrophosphate dihydrate crystals deposition disease (chondrocalcinosis), associated with osteoporosis. Hemochromatosis is the clinical condition of iron overload; it can be genetic or secondary. It is important to know how to diagnose early genetic hemochromatosis since it is a frequent, life-threatening and treatable condition. Early bio clinical symptoms are weakness, arthralgias and increase of serum tranferrin saturation. PMID- 17416052 TI - [Post transfusionnal iron overload]. AB - Post transfusionnal iron overload is related to both a degree of RBC units transfused and excess intestinal absorption of Fe related to dyserythopoiesis. This condition is associated with high rates of morbidity and mortality, mainly of cardiac origin. Iron chelation therapy in patients with thalassemia who were effectively chelated has prevented or partially reversed some of these consequences. However the reference treatment by deferoxamine is strenuous and compliance is poor. Two oral iron chelators, deferiprone and deferasirox, provide potentially useful treatment for iron overload, both agents are relatively well tolerated (mainly deferasirox). They were at least as effective as DFO for decreasing iron burdens in comparative trials, and were associated with improved cardiac outcomes (for deferiprone). A new enthusiastic area for iron chelation with less burdensome treatments is open; however a long term follow up is required before giving up pumps and needles. PMID- 17416053 TI - [Patient's record. Treatment of hemochromatosis]. PMID- 17416054 TI - [Faith and ten commands]. PMID- 17416055 TI - [Psoriasis]. PMID- 17416056 TI - [Acute and chronic headaches]. PMID- 17416057 TI - [Aortic stenosis]. PMID- 17416058 TI - [Vomiting in adults (with treatment)]. PMID- 17416059 TI - [Normal ageing: biological, functional and relational aspects. Epidemiological and sociological date. Prevention of the pathological ageing]. PMID- 17416060 TI - [The revolving solarium of Aix-les-Bains (France)]. PMID- 17416061 TI - Analysis of the filamentous bacteriophage genomes integrated into Neisseria gonorrhoeae FA1090 chromosome. AB - Bioinformatic analysis of the genome sequence of Neisseria gonorrhoeae revealed presence of four specific prophage islands. Based on the similarity with other DNA phage sequences they seem to belong to the filamentous ssDNA phages group. Phages belonging to this group are also present in the genome of Neisseria meningitidis. The nucleotide and amino acids sequence of Ngo phi6 and Ngo phi7 show similar genetic organization and high homology on DNA and amino acid level. The Ngo phi9 contains only part of the genomes of the Ngo phi6-8 prophages. Several functionally same genes of different origin are duplicated, with no homology to their counterparts in phages Ngo phi6, Ngo phi7 and Ngo phi9. The prophage sequences of nucleotides of Ngo phi6 and Ngo phi7 contain specific blocks of genes responsible for phage DNA replication and structural proteins. Comparative analysis at nucleotide and amino acid level suggests that these sequences can encode functionally active phages. The genetic organization of the Ngo phi6 suggests that it can serve as a prototype of filamentous phage of N. gonorrhoeae. Presence of the genomic ssDNA of these phages in the cultures of N. gonorrhoeae confirms this conclusion. PMID- 17416062 TI - Replication system of plasmid pMTH4 of Paracoccus methylutens DM12 contains an enhancer. AB - The replication system of plasmid pMTH4 (22 kb) of dichloromethane-degrading Paracoccus methylutens DM12 (Alphaproteobacteria) has been cloned within a mini replicon pMTH100 (4.7 kb) and preliminarily characterized. Functional analysis, performed with a series of mutated plasmid mini-derivatives, showed that the replicator region consists of three elements: (i) gene repA coding for a replication initiation protein RepA, (ii) origin of replication (oriV), placed in the promoter region of repA and containing a set of imperfect directly repeated sequences (iterons) together with putative DnaA and IHF-binding DNA sequences as well as (iii) an enhancer (0.65 kb) upstream of oriV. We showed that the enhancer was not crucial for plasmid replication, however, it was necessary to assure the proper plasmid copy number. Additionally its presence has increased the strength of a determinant of incompatibility (located within the oriV region) as well as the level of transcription carried from the repA promoter. The enhancer region was shown not to encode any proteins or promoter sequences. We speculate that this region might constitute a site of binding of plasmid or host-encoded proteins that are able to interact with the origin, which positively regulates the initiation of replication. PMID- 17416063 TI - Acquisition of iron by enterococci: some properties and role of assimilating ferric iron reductases. AB - Ferric iron reductases activities have been occurred in 91% of investigated enterococci strains. Maximum activity occurred with coenzyme NADH as the reductant and the presence of cofactor FMN was necessary. Mg(II) ions has not stimulated reductases activity. Treatment of cells with proteolytic enzymes had not effect on iron reduction. The whole cells and cell fraction-cytoplasmic membrane and cytoplasm showed Fe(III)-reducing activity. The highest specific activity was associated with cytoplasm. The activity in cytoplasmic membrane was not related to iron concentration in the growth medium. In cytoplasm the activity was stimulated after growth in low-iron medium. Ferric iron reductases of enterococci characterized the broad substrate specificity. The iron in form of ferric ammonium citrate, lactoferrin and ferrioxamine B were the best iron sources for enterococcal ferric iron reductases. PMID- 17416064 TI - Susceptibility of Listeria monocytogenes strains isolated from dairy products and frozen vegetables to antibiotics inhibiting murein synthesis and to disinfectants. AB - The susceptibility of 96 strains of Listeria monocytogenes isolated from food to antibiotics and disinfectants currently used in human therapy, veterinary, medicine and food industry was determined by a standard operating procedure- broth dilution method. Antimicrobial agents included the beta-lactams ampicillin and penicillin, the lantibiotic nisin, and the disinfectants benzalkonium chloride and chlorhexidine gluconate. Among the studied strains we found 13 strains with 8-fold, 7 strains with 16-fold and 2 strains with 32-fold decreased susceptibility to ampicillin, as determined by MIC, compared to wild type reference strain. Interestingly, the mentioned strains were isolated from frozen vegetables and soups, none of the isolates from dairy products showed any elevated resistance to the studied antimicrobial agents. The occurrence in food products of strains with increased resistance to ampicillin is disquieting, especially since 3-lactams are the most frequent antibiotic of choice in the therapy of infections caused by the pathogen. PMID- 17416065 TI - Partial characterization and optimization of production of extracellular alpha amylase from Bacillus subtilis isolated from culturable cow dung microflora. AB - Studies of alpha-amylase production by Bacillus subtilis (CM3) isolated earlier from cow dung microflora, were carried out. The optimum temperature, pH and incubation period for amylase production were 50-70 degrees C, 5.0-9.0 and 36 h, respectively. Enzyme secretion was very similar in the presence of any of the carbon sources tested (soluble starch, potato starch, cassava starch, wheat flour, glucose, fructose, etc.). Yeast extract and ammonium acetate (1%) as nitrogen sources gave higher yield compared to other nitrogen sources (peptone, malt extract, casein, asparagine, glycine, beef extract), whereas ammonium chloride, ammonium sulphate and urea inhibited the enzyme activity. Addition of Ca+2 (10-40 mM) to the culture medium did not result in further improvement of enzyme production, whereas the addition of surfactants (Tween 20, Tween 40, Tween 80, and sodium lauryl sulphate) at 0.02% resulted in 2-15% increase in enzyme production. There were no significant variations in enzyme yield between shaked flask and laboratory fermentor cultures. The purified enzyme is in two forms with molecular mass of 18.0 +/- 1 and 43.0 +/- 1 kDa in native SDS-PAGE. PMID- 17416066 TI - Production of tannase through submerged fermentation of tannin-containing plant extracts by Bacillus licheniformis KBR6. AB - Tannins are water-soluble polyphenolic compounds found in plants as secondary metabolites. The presence of these substances in the barks of eight different plants was initially examined and their crude extracts were used separately as a substrate for production of tannase through submerged fermentation by Bacillus licheniformis KBR6. Tannase production as well as biodegradation of the substrate reached a maximum within 15 to 18 h against crude tannin extract obtained from Anacardium occidentale. Among different concentrations of the crude tannin tested, 0.5% (w/v) induced maximum synthesis of enzyme. Tannase production was higher by almost two-fold in the presence of crude tannin compared to pure tannic acid used as a substrate. It seems that industrial production of tannase, using bark extract of A. occidentale can be a very simple and suitable alternative to presently used procedures. PMID- 17416067 TI - In vitro activity of synthetic antimicrobial peptides against Candida. AB - Yeast-like fungi are the most common cause of fungal infections in humans. Actually, in the age of opportunistic infections and increasing resistance, development of modern antifungal agents becomes a very important challenge. This paper describes synthesis and antimicrobial assay of four naturally occurring peptide antibiotics (aurein 1.2, citropin 1.1, temporin A, uperin 3.6) and three chemically engineered analogues actually passing clinical trials (iseganan, pexiganan, omiganan) against Candida strains isolated from patients with infections of the oral cavity or respiratory tract. The peptides were synthesized using solid-phase method and purified by high-performance liquid chromatography. Biological tests were performed using the broth microdilution method. The antifungal activity of the peptide antibiotics was compared to that of nystatin and amphotericin B. We found synthetic peptides to be generally less potent than amphotericin B or nystatin. However, some of the naturally occurring peptides still retained reasonable antifungal activities which were higher than these of iseganan, pexiganan or omiganan. We think that the naturally occurring peptide antibiotics included in our study can be a good matrix for development of novel antifungal compounds. PMID- 17416068 TI - Effects of culture conditions on production of extracellular laccase by Rhizoctonia praticola. AB - It was found that the soil-dwelling fungus Rhizoctonia praticola 93A was capable to produce laccase in submerged cultures. Effects of culture conditions on the enzyme biosynthesis in shaken flask and aerated bioreactor cultures were evaluated to improve the yields of the process. Production of extracellular laccase was considerably intensified by the addition of Cu2+ to a carbon-limited and nitrogen-sufficient culture medium (C/N = 0.98). When an optimized medium containing glucose (2 g/l) and L-asparagine (1.5 g/l) was used and enzyme synthesis was stimulated by addition of 5 microM Cu2+ before inoculation, maximal laccase activities obtained in a batch cultivation were, approximately, 1000 nkat/l. Under these conditions, addition to the medium of the aromatic inducer 2,5-xylidine (1 mM) led to a 10-fold increase in laccase activity. Laccase productivity in shaken flask cultures was also enhanced (to more than 4000 nkat/l on day 3) by using a medium with the initial pH of 7.5. Such a high value of the optimal medium pH for laccase production by R. praticola is exceptional among the ligninolytic fungi. In fermenter fungal cultures supplemented with cupric ions, the highest laccase activity (about 4000 nkat/l after 3 days' cultivation) was reached after 24-h incubation using a bioreactor with the aeration rate of 21/min, the agitation speed of 200 rpm, and a constant medium pH of 8.0. PMID- 17416069 TI - Growth of Penicillium verrucosum and production of ochratoxin A on nonsterilized wheat grain incubated at different temperatures and water content. AB - The results of two experiments with wheat grain inoculated with Penicillium verrucosum are reported. In Experiment I, wheat grain, containing 10, 20 and 30% water, was incubated for 2 weeks at 10, 15, 21 and 28 degrees C. In Experiment II, wheat grain, containing 14, 16, 18, 20 and 22% water, was incubated for 2 weeks at 10, 15, and 20 degrees C. At initial moisture content (IMC) of the wheat grain up to 16% neither P. verrucosum growth nor ochratoxin A (OTA) formation were observed. In the range of IMC from 18% to 22% both the fungal growth and OTA synthesis were distinct, and the parameters were higher at higher temperature in the range 10-21 degrees C. A temperature of 28 degrees C was probably too high for proper metabolism of the fungus, including OTA formation. OTA formation was distinctly related to P. verrucosum abundance in the temperature range 10-21 degrees C, expressed both as the counts of fungal colony forming units (CFU) on agar DYSG medium and diameters of the fungal colonies growing around the wheat kernels placed on the surface of DYSG medium. OTA formation and abundance of P. verrucosum were negatively correlated with the percentage of wheat kernels, placed on DYSG medium, with growing colonies of fungi different from P. verrucosum. CFU counts of P. verrucosum on the wheat grain were significantly related to the diameter of the fungal colonies growing around the wheat kernels placed on DYSG medium. The relationship is described by an exponential regression equation. PMID- 17416070 TI - Detection of cytomegalovirus and Helicobacter pylori DNA in arterial walls with grade III atherosclerosis by PCR. AB - It has been suggested that some microorganisms may play a role in the etiology or progression of atherosclerotic plaques. The purpose of this study was to assess for the presence of Helicobacter pylori and cytomegalovirus (CMV) DNA using polymerase chain reaction (PCR) technique in vascular-wall specimens obtained during autopsy. Four to 5 mm long samples from 3 different vascular wall specimens (coronary, carotid and abdominal aortas) of 30 patients (23 male, 7 female) were taken for pathologic and microbiologic investigations during autopsy. H. pylori DNA was found in 48.2% atherosclerotic and 19.6% non atherosclerotic vascular wall specimens, whereas CMV DNA was found in 37.9% atherosclerotic and 32.7% non-atherosclerotic vascular wall specimens. In terms of CMV DNA detection, no statistically significant differences between the atherosclerotic and non-atherosclerotic groups were present (P > 0.05). However, there was a statistically significant difference between the atherosclerosis and non-atherosclerotic groups in terms of H. pylori DNA in coronary and abdominal aorta arteries (p = 0.016 and p = 0.0029 respectively) but not in carotid arteries (p = 1.00). In conclusion, the correlation between H. pylori and atherosclerosis could be suggested. These finding warrant further investigation regarding the role of H. pylori in atherosclerosis. PMID- 17416071 TI - Detection of methicillin resistance in hospital environmental strains of coagulase-negative staphylococci. AB - The aim of this study was to evaluate methicillin resistance detection methods currently used when studying coagulase-negative staphylococci (CONS). The resistance to oxacillin of 142 strains from seven species of CoNS isolated from the Intensive Care Unit environments was tested. The methods used were: disc diffusion test with cefoxitin (FOX30) and oxacillin (OX1), oxacillin agar screen test with 6 mg/l of oxacillin (MHOXA), latex test for PBP2a (LA) and detection of mecA via PCR. One hundred and one isolates were methicillin-resistant in at least one of methods used, but only 74 were mecA-positive. Of the 68 mecA-negative strains: two were positive by OX1, the LA and MHOXA methods; three by the LA and MHOXA; and 22 only by OX1 test. Most of these strains were from the novobiocin resistant CoNS group. The results obtained for all tested strains using FOX30 showed complete concordance with the presence of the mecA gene. PMID- 17416072 TI - [Subrenal aorta resection procedure in dissections of the thoracic aorta using fenestration technique]. AB - Three cases of dissections of the thoracic aorta involving pelvic vasculature were managed in the Plzen surgical clinic during 2004-2005, using aortic fenestration technique. In one case, the procedure was urgent due to acute ischemia of an extremity. The other two procedures were planned procedures. The outcome of the three procedures was successful and no treatment complications were recorded. PMID- 17416073 TI - [The incidence of postoperative residual curarization in the recovery room after rocuronium administration]. AB - In 513 patients, we investigated residual curarization after general anaesthesia with rocuronium administered, without peroperative neuromuscular blockade monitoring. On admission to the recovery room, the ulnar nerve was stimulated submaximally (30 mA) and the evoked muscle response was quantified with accelerometry (TOF-Watch SX, Organon). The postoperative residual curarization was defined as a TOF-ratio < 0.9 and could be demonstrated in 174 patients (34 %). Compared to the group with adequate recovery, these patients received larger rocuronium dose [45.4 (SD 13.2) mg vs. 40.4 (SD 14.3) mg, p < 0.01], less experienced anaesthesiologists conducted their case [p <0.01], shorter time had elapsed since the last rocuronium dose [58.4 (20.9) min. vs. 64.9 (27.2) min., p < 0.05], their core temperature was lower [35.4 (0.6) degrees C vs. 35.8 (0.6) degrees C, p < 0.011 and on average, they received less neostigmine during anaesthesia [0.26 (0.47) mg vs. 0.57 (0.71) mg, p < 0.01]. We conclude that it is necessary to antagonize residual block after rocuronium, especially in the absence of perioperative neuromuscular monitoring. PMID- 17416074 TI - [Insufficient anastomoses in sigmoideal and rectal resections. A retrospective study conducted in a surgical clinic in Hradec Kralove]. AB - INTRODUCTION: Anastomotic insufficiency is the primary cause of postoperative morbidity and mortality following resection procedures of the large intestine and rectum. MATERIAL AND METHODOLOGY: In the retrostpective study, the authors analysed rates of rectal and sigmoideal anastomotic insufficiencies in patients operated for rectal and sigmoideal carcinomas in the Faculty Hospital Surgical Clinic in Hradec Kralove from 2000 to 2004. At the same time, the authors analysed risk factors of the insufficiencies. The subject of protective derivation stomies is discussed. RESULTS: In the group with primary colorectal anastomosis, the anastomotic insufficiency occured in 11% of the group subjects, in the group with primary sigmoideal anastomosis in 9.1% of the group subjects. Out of the total of 215 subjects, the anastomotic insufficiency occurred in 23 subjects (10.7 %), 6 cases were fatal and the overall postoperative mortality was 1.56 % . In the anastomotic insufficiency group, it reached 13.04 %. The difference between the studied groups is significant (p<0,001, OR = 10.5). CONCLUSION: Postoperative mortality in sigmoideal and rectal resection procedures correlates with anastomotic insufficiency. PMID- 17416075 TI - [The Peutz-Jeghers syndrome--a case review]. AB - INTRODUCTION: The authors present a case review of a localized Peutz-Jeghers syndrome of the caecum, emulating a carcinoma. METHODS: The patient who presented with insignificant family history with a diagnosed stenosing tumor of his appendix and histological findings of highly suspected adenocarcinoma, was indicated for surgical revision and right-sided hemicolectomy. RESULTS: The patient underwent laparoscopically assissted right-sided hemicolectomy and his postoperative course was adequate. The final histological finding diagnosed solitary hamartoma of the Peutz-Jeghers polyp. CONCLUSION: Differential diagnostic reasoning in patients with negative family history and unclear case history with findings of polypous tumorous lesions of the GIT, should also consider the Peutz-Jeghers syndrome. The syndrome is related to a high risk of malignancies in the elderly. PMID- 17416076 TI - [Liver hemangiomas--diagnostic and treatment strategy]. AB - Liver hemangiomas are benign mesenchymal tumors or the liver. The majority of them are asymptomatic, diagnosed accidentally during abdominal examination for other reasons. Ultrasonography. computer tomography and magnetic resonance are the main diagnostic instruments. Symptomatic, large cavernous hemangiomas or hemangiomas located on the liver surface with a risk of rupturing because of their size, are indicated for surgical management or radiointervetion. The authors present a patient group of 34 subjects with cavernous hemangiomas of the liver indicated for surgery (N = 25), or radiointervention (N = 9), based on the above critera. The thirty-day mortality rate was nil and morbididty 5.9%. The authors recommend enucleation of the hemangioma, which is technically simple and safe. From a long-term perspective, radiointervention is frequently poorly effective, and, therefore, should be indicated in polymorbid patients. PMID- 17416077 TI - [Laparoscopic correction of posttraumatic uterovaginal descent with rectal prolapse]. AB - A case review of laparoscopic management of the uterovaginal descent and rectal prolaps in one step in a young female following her complicated pelvic fracture. PMID- 17416078 TI - [Multiple diverticulosis of the small and large intestine. A case review]. AB - The authors present a case review of a patient with relapsing diverticulosis of the large and small intestine. They describe surgical strategy in the patient and discuss ethiopathogenesis of the diverticular disorder, as well as options and extent of the surgical treatment. PMID- 17416079 TI - [The role of CT for indicating laparotomy in blunt abdominal trauma: comparing CT results and surgical findings in a group of 101 patients. Prospective study]. AB - BACKGROUND: The helical contrast CT plays an important role in the management of abdominal trauma. For surgeons its reliability and utilization remain questionable. OBJECTIVE: The study aimed at correlating CT results at admission with surgical findings in patients undergoing laparotomy for blunt abdominal trauma, detecting CT sensitivity for therapeutic laparotomy and uncovering organ injury. The group of patients is from one hospital; therefore mistakes from different result interpretations or data collecting are not present. METHOD: A rentgenologist's conclusion during patient admission, late independent CT evaluation by an experienced rentgenologist and surgical finding were statistically correlated. RESULTS: 101 out of 139 patients underwent laparotomy after initial CT. Mean ISS were 27.8. CT sensitivity (SE) for indicating therapeutic laparotomy was 88.77/ 89.88%, predictive value positive result (PVPR) 92.94/94.11%, and false negativity (FN) 11.23/10.11 %. SE in liver and spleen trauma was 88.76/95.50%, PVPR 96.34/93.41%, and FN 11.24%/ 4.50%. SE in bowel and mesenteric injury was 27.27/18.18 %, SE in diaphragm injury was 33.33 %. CONCLUSION: the study presented high SE of CT forecasting therapeutic laparotomy for blunt abdominal trauma. The high SE of CT in liver and spleen injuries is useful in indicating conservative therapy. The CT accuracy was not confirmed in bowel. mesenteric or diaphragm injury. It strongly recommend patient observation even with negative CT. PMID- 17416080 TI - [Undiagnosed posterior dislocations of the shoulder joint. A preliminary report]. AB - Posterior dislocations are infrequent injuries of the proximal humerus, counting for 1-4% of all shoulder dislocations. Frequently, they are related to epileptic seisures or electric current injuries or accompany complete anatomical neck fractures of the proximal humerus. Very frequently, they remain undiagnosed and their late diagnosis results in poor functional outcomes. The authors assess the surgical procedure and its results in five cases of primarily undiagnosed posterior dislocations. In two cases, transposition of the m. subscapularis was performed, in one case hemiarthroplasty, and in two cases derotation osteotomy of the proximal humerus was used. Although the study group was small, the preliminary results of the procedures for chronic posterior dislocation of the shoulder joint favours preservative procedures compared to hemiarthroplasty of the shoulder joint. PMID- 17416081 TI - [Opportunity or frustration in social services: a law for dependence]. PMID- 17416082 TI - [Adolescent adjustments. The social support perspective]. AB - In this article, the authors analyze the relationship which exists among different sources of social support and three health indicators during adolescence: the presence of depressions, alcohol consumption and smoking cannabis. The sources of social support analyzed are: support by the father support by the mother; support by one's steady boy/girl friend, and support by one's best friend. The sample for this study is comprised of 1002 adolescents aged 12 to 20. Results show that a greater perception of support by the father and by the mother leads to lesser incidences of depression. The perception of greater paternal support is also connected to lesser consumption of both alcohol and cannabis. However the perception of greater support by one's steady boy/girlfriend is associated with a higher degree of alcoholic consumption during adolescence. PMID- 17416083 TI - [Respiratory therapy and essential hypertension (II). An incidence study]. AB - The authors carried out a study on the occurrence of breathing controlled by respiratory cycles comprised by short inhalations and long exhalations. The results showed a significant decrease regards the base line parameter for both systolic and diastolic arterial pressure at the end of treatment which had 8 sessions, at six months and at two years. The systolic and diastolic values fall within the recommended WHO values in 1999. To receive more extensive information regarding materials, procedures and results, please contact the authors. PMID- 17416084 TI - [Neonatal screening for congenital metabolic pathologies. Ethical and social aspects. Implications for nurses and midwives]. AB - The authors describe the diseases which are detectable by means of neonatal screening programs available in the different Spanish Autonomous Communities. They analyze some of the ethical and social implications which may occur and they emphasize what those programs mean for nursing professionals, fundamentally midwives and nurses who work in primary health care units or pediatric units. PMID- 17416085 TI - [Pain during venipuncture. The efficacy of physical contact]. AB - The authors show the relationship between the use of physical contact and the decrease in pain during venipuncture, or blood-drawing, by means of a clinical test and an experimental, random, controlled study. Random sample by means of a systematic probability sampling. Two groups, control and experimental, were set up, employing physical contact in the latter group. To measure pain, an Analogical Visual Scale from 0-10 (EVA 0-10) was used and a data-register sheet was developed. Pain decreased by 37.45% (p = 0.000) in the experimental group, one of the main objectives of this study. PMID- 17416086 TI - [Mistreatment: detection of mistreatment in a sample feminine population]. AB - The authors identify possible psychological mistreatment and symptoms of anxiety and depression in a sample of women who were attended to in the "Sant Josep" Primary Health Care Center in L'Hospitalet de Llobregat in Barcelona Province between February and June 2005 by means of an observational, descriptive and transversal study with a consecutive study of women older than 18 who were attended to by nurses for whatever health problem they had or who filed administrative paperwork. Data was obtained through the use of a self administering questionnaire on which these items were recorded: social-health variables, symptoms of anxiety and depression, and questions elaborated ad hoc in order to identify mistreatment. More than half of the study participants complied with the criteria used to indicate symptoms of anxiety and depression, which recommended a follow-up session. The authors observed an association between depression and mistreatment. Part of the results of this study were presented in The 1st World Congress on Gender-Specific Medicine in February 2006 in Berlin. Financial Sources: The School of Nursing at the University of Barcelona (PREU12004/10) and the Catalan Institute for Women (U-39/05). PMID- 17416087 TI - [Risk of a fall in a critical hospital. The cognoscible state and the dependence level]. AB - Falls by the elderly are an important cause of fatalities, they tend to increase hospital stays and they increase health care payments and family expenses. Consequently falls have an adverse effect which requires their prevention by identifying risk factors and establishing preventive measures. Given the importance of this type of accident, the authors attempt to discover if there is a relationship between a fall and the cognoscible state of those patients -to carry out this evaluation, the "Lobo" test is applied- and the fall and the degree of independence a patient has by means of the Barthel test. The results obtained show that, compared to the control group, patients included in the "GDR" sector (Diagnostically Related Groups) of the central nervous system and the GDR of the locomotive apparatus suffered more falls and therefore, in consequence, the factors which predict a fall have more relationship with a patient's basic disease than with the number of clinical processes which affect a patient. As opposed to our initial idea risk of fall factors, we found no association to lower values on the Lobo nor Barthel scales; which is to say that functional dependencies or cognoscible deterioration do not lead to more falls. PMID- 17416088 TI - [HIV sexual transmission. Sexuality and at risk behavior]. AB - At the present time, HIV infection occurs mainly as a sexually transmitted disease everywhere in the world as 75-80% HIV contagion occurs by this means. Although 40-70% of cases of HIV infection continues to occur among young male homosexuals, the annual incidence in new cases by which HIV is transmitted through heterosexual contact is undergoing a constant increase throughout the world, especially among females; 40% in males and close to 80% of the new infections by heterosexual contact in females. The findings make obvious the need to insist on preventive measures as the only way to stop this means of HIV transmission. PMID- 17416089 TI - [Controversy over one use only medical devices]. AB - At the present time, concern has arisen as to whether or not medical devices designed for one use only should be reutilized after the appearances of outbreaks of Hepatitis C infections in Spain as well as in other countries. 80% of Spanish hospitals, both public and private, reprocess these one use only devices; there is no regulation at this time which guarantees this practice. The increase in endoscopic surgeries has led to an increase in health care costs, plus an increase in the residues which derive from their use. This fact has caused the reutilization of one use only devices in order to balance costs. According to a study in the United States, the price of a reprocessed one use only device is approximately half the price of a new one, and at times, the cost of these reprocessed devices is less than 10% that of a new one. In our country, a precise classification regarding which materials may or may not be reused does not exist, nor do protocols for sterilization and reprocessing and reuse of one use only devices. An agreement should be made by the parties involved; the administration, sanitary professionals, manufacturers of medical devices and the users which guarantees a balance between the important increase in health care costs due to throwing away these products and the insecurity which their use without a ratified guarantee means. PMID- 17416090 TI - [Therapeutic strategies for acute type A aortic dissection]. AB - We herein present the early and mid-term outcomes of therapeutic strategies for acute type A aortic dissection in our department. Subjects were 75 patients who were admitted to our department from January 2001 to October 2006. A total of 33 patients had thrombosed dissection: emergent surgery was indicated for cases in which the maximal ascending aortic diameter was > or =50 mm or when ulcer-like projection (ULP) was observed in the ascending aorta. Only 1 case of rupture related death was observed, in a patient who had a maximal ascending aortic diameter of 52 mm and refused surgery. Although 8 patients were converted to surgery during the chronic phase, elective surgery was recommended in all cases. Surgery consisted of entry resection using open distal anastomosis under circulatory arrest at a bladder temperature of 25 degrees C, with antegrade cerebral perfusion into the 3 cervical branches of arch aorta based on bilateral axillary artery. In-hospital mortality of the 62 patients who underwent surgery was low (4.8%) and no dissection-related deaths were reported for the midterm outcomes. In addition, a low rate of cerebral infarction was observed among cases who had residual dissection of the brachiocephalic arteries after surgery. These findings demonstrate the validity of the therapeutic strategies in our department. PMID- 17416091 TI - [Surgical treatment for acute type A aortic dissection]. AB - The purpose of this study was to assess the factors for clinical outcome of the surgical treatment of acute type A aortic dissection. From April 1996 to March 2006, 44 patients underwent emergency operation for acute type A dissection within 2 weeks from the onset. Resection of the intimal tear was performad with open distal anastomosis. The mean age was 63.4 (range 29-83) years, and 28 were female. As for their preoperative condition, 5 patients were in severe hemodynamic instability including cardiac arrest in 2, apnea in 1, and rupture in 4. Distal resection extended to ascending aorta in 24 patients (54.5%), hemiarch in 7 (15.9%), and total arch in 13 (29.5%). 30-day mortality was 4.5% and the incidence of stroke was 13.6%. Several methods were used including axillary artery cannulation and central repair with adventitial inversion technique. Patients with malperfusions caused by acute type A dissection should undergo immediate aortic reconstruction by adequate circulatory assisting methods. PMID- 17416092 TI - [Surgical treatment for acute type A aortic dissection; for better postoperative quality of life]. AB - From November 1999 to December in 2005, 114 patients with acute type A aortic dissection underwent surgical treatment on an emergency basis. The overall in hospital mortality was 7.9% (9 patients). Four were rupture cases before cardiopulmonary bypass. De novo postoperative stroke rate was 3.5% (4 patients). But all of them were discharged on foot. There were 6 rupture cases before operation. Unfortunately only 2 patients survived. Preoperative stroke due to malperfusion occurred in 19 cases (16.7%). Among them, those with clear consciousness had tendency to better social rehabilitation than those with drowsiness. We had experienced 2 vegetable states in the group of drowsiness after the operations. For better outcome, we must avoid rupture before operation and reconsider the timing of operation in the case of brain ischemia. PMID- 17416093 TI - [Early and late sugical results of emergency operation for acute type A aortic dissection]. AB - We evaluated the results of 213 emergency operations of acute type A aortic dissection in our center from January 2003 to December 2006. They were 101 male and 112 female, and the mean age was 64.6 years. The hospital mortality rate of all cases was 13.6% (29/213). And that of cases with malperfusion was 31.9% (15/47). They consisted of stroke 8/17 (47.1%), myocardial ischemia 5/27 (18.5%) [right coronary artery: 2/22 (9.1%), left main trunk: 3/5 (60.0%)], and intestinal ischemia 2/3 (66.7%). The hospital mortality rate of pulseless electrical activity (PEA) cases was 57.1% (4/7), and that of aortic rupture cases was 33.3% (3/9). On the other hand, the mortality rate of cases with cardiac tamponade alone was 4/45 (8.9%). That of cases without cardiac tamponade and malperfusion was 3/105 (2.9%), and was significantly (p < 0.05) lower than that with malperfusion. PMID- 17416094 TI - [Is delayed surgical repair admitted for the thrombosed aortic dissection?]. AB - The initial treatment for the thrombosed aortic dissection is still controversial. Accordingly, we sought to evaluate the strategy of its surgical repair. Ninety-six (35 type A and 61 type B) acute thrombosed aortic dissection were studied retrospectively. Initially all of them were treated medically. The ratio of the false and the true lumen (F/T ratio) was calculated on the onset. The uncontrollable cardiac tamponade, recanalization, large ulcer-like projection (ULP) and enlargement of the dissected aorta had a delayed surgical repair during the follow-up period. Eighteen of the type A and 14 of the type B were surgically treated and showed good result. The 1- and 5-year survival rates and the event free survival rates for the type A and the type B were almost equal with no statistical difference. The mean F/T ratio for the type A was 31% for the operative and 51% of the nonoperative cases (p = 0.007). The maximum size of the initial aorta of the operative cases was larger than that in the nonoperative for the type B. The conservative therapy for the thrombosed aortic dissection decreased the number of the unnecessary operation and the strategy of the delayed surgical repair was justified properly. PMID- 17416095 TI - [Surgical strategy for acute type A aortic dissection with cerebral ischemia due to acute occlusion of aortic arch branches]. AB - In the treatment of acute type A aortic dissection, it is important to cope effectively with cerebral ischemia due to preoperative acute occlusion of arch branches and intraoperative cerebral malperfusion under extracorporeal circulation. The validity of our surgical strategy for such cases was evaluated. Our surgical strategies are as follow; for cases with preoperative cerebral infarction and disturbance of consciousness total aortic arch replacement is performed after the improvement of brain edema, and for cases of transient cerebral ischemia, emergency operation is performed. In the emergency operation, selective cerebral perfusion through the carotid artery of the diseased side is initiated as soon as possible. In conclusion, our surgical strategy for acute type A aortic dissection with cerebral ischemia due to acute occlusion of aortic arch branches is acceptable. There was no significant difference between the cerebral ischemia group and the control group concerning hospital mortality, cerebral complication and the 5-year survival rate. PMID- 17416096 TI - [Coronary malperfusion due to acute type A aortic dissection; surgical strategy and results]. AB - Coronary malperfusion due to acute type A aortic dissection (DAA) is a lethal complication. It is especially difficult to rescue the patients with left coronary malperfusion because of acute global myocardial infarction (AMI), even with successful surgical treatments, including the replacement of the ascending aorta and coronary artery bypass grafting (CABG). We review our experience and illustrate our approach to these critically ill patients. In addition, we classify the mechanism of malperfusion into 4 types based upon perioperative findings and discuss surgical management indivisually. From January 1990 to April 2005, a total of 260 patients were operated for DAA in our institution. Twenty (7.7%) patients, 11 men and 9 women were suffering from coronary malperfusion due to DAA. The mean age was 55 (range 28-72) years. The right coronary artery was involved in 9 patients, and the left in 11. All procedures such as graft replacement and CABG were done on an emergent or urgent basis. Hospital mortality rate of right coronary malperfusion was 22% (2/9 patients), and that related to left coronary malperfusion was 5/11 (45%). Assisting device was required in 9 cases, veno-arterial bypass (VAB) in 6 cases, left ventricular assist system (LVAS) in 1, left heart bypass (LHB) in 1, LHB+right heart bypass (RHB) in 1. We lost all patients using VAB. Only 3 patients supported with strong assist device survived. Aggressive myocardial resuscitation and early operation are the key factors in the management of these critically ill patients. But once severe myocardial infarction occurs, V-A bypass (percutaneous cardiopulmonary support) is useless in treating patients with DAA who develop severe heart failure. We recommend to implant stronger assist device including LVAS immediately before exacerbation of multiple organ failure. In conclusion, surgical management is not easy for emergency patients with DAA in association with myocardial ischemia. However, reasonable surgical results can be obtained with supplemental CABG and strong mechanical support of the left ventricle. PMID- 17416097 TI - [Treatment of acute type B aortic dissection with organ ischemia]. AB - Nine patients, who suffered from acute type B aortic dissection with organ ischemia, were treated at our hospital from 2004 to 2006. Their mean age was 60.3 (range 37-73) years. Eight of them required surgical intervention. Two patients with mesenteric-ischemia underwent superior mesenteric artery (SMA) bypass surgery and their conditions were relieved. However, 1 of them died of aortic rupture 6 months later. One patient with celiac artery occlusion was at first treated nonsurgically, but was subjected to resection of the small intestine 3 weeks later because of ulcer perforation induced by ischemia. The other 5 patients with lower extremity ischemia underwent bypass surgery and were discharged. Bypass surgery is a reliable procedure for the treatment of acute type B aortic dissection with organ ischemia, allowing prompt resolution of ischemia. PMID- 17416098 TI - [Treatment strategy for acute type A aortic dissection; how to manage mesenteric ischemia]. AB - Mesenteric ischemia is a dreaded complication of acute type A aortic dissection. From January 1994 to December 2004, 134 patients with acute type A aortic dissection were operated. Eleven patients showed postoperative mesenteric ischemia. Mortality of such patients was much higher than that without mesenteric ischemia (81.8 vs. 10.6% , p < 0.0001). Preoperative mesenteric and/or lower extremity ischemia were revealed to be the risk factors of postoperative mesenteric ischemia. Our strategy to manage these patients is as follows; patients who are suffering mesenteric and/or lower extremity ischemia preoperatively, or those whose computed tomography (CT) shows stenosis, obstruction, or dissection of the superior mesenteric artery, should be recognized as high-risk patients of postoperative mesenteric ischemia. Their mesenteric circulation should be examined directly with laparotomy after the central repair. If the mesenteric circulation seems to be suboptimal, iliac artery-superior mesenteric artery bypass should be performed. PMID- 17416099 TI - [Transapical aortic cannulation in the operation of acute type A aortic dissection]. AB - From June 2003 to November 2006, transapical aortic cannulation was performed in 73 patients (41 men and 32 women, mean age 63 years, 64 hemiarch repair and 9 total arch replacement) with acute type A aortic dissection. A 1-cm incision was made in the apex of the left ventricle, and a 7-mm soft and flexible cannula was passed through the apex and across the aortic valve until positioned in the ascending aorta under guidance by transesophageal echocardiography. In all cases, cardiopulmonary bypass flow was sufficient. There were no malperfusion events. Our results showed that transapical aortic cannulation was secure and useful for repair of acute type A aortic dissection. PMID- 17416100 TI - [The strategy to avoid malperfusion of vital organs during operation for acute aortic dissection; multi-monitoring and right brachial artery perfusion]. AB - BACKGROUND: Malperfusion of vital organs is the most serious complication during cardiopulmonary bypass for acute aortic dissection. METHOD: From 2001 to 2006, 40 patients underwent operation for acute type A aortic dissection. Right brachial artery perfusion was performed in 20 patients. From May 2005, transesophageal echocardiography and cerebral oxygenation measured by near infrared spectroscopy were continuously monitored during operation. RESULTS: There were 3 in-hospital deaths and 3 brain infarction. Switching of blood flow during cardiopulmonary bypass occurred in 3 patients; 2 patients suffered from diffuse brain infarction while. in the other patient, switching of perfusion was detected at once by multi monitoring and resolved by induction of right brachial artery perfusion. CONCLUSION: Close monitoring of cerebral oxygenation by near infrared spectroscopy, transesophageal echocardiography and right brachial artery perfusion are effective in operation for acute aortic dissection. PMID- 17416101 TI - [Durability of aortic root preservation using gelatin-resorcin-formalin glue in acute type A aortic dissection]. AB - During the last 9 years, aortic root preservation using gelatin-resorcin-formalin (GRF) glue was performed in 63 patients as a part of surgery for acute type A aortic dissection. Residual aortic regurgitation (AR) was evaluated, grading 0 to IV+ by echocardiography. The survival and root reoperation-free rates were also assessed. The operative mortality was 9.5% (6 patients). Early postoperative AR < or = I+, = II+ and > or =III+ were 93, 7 and 0%, respectively. Late postoperative AR > or =III+ was observed in 4 patients. Root reoperation was performed in 4 patients (7.0%). In a case of reoperation, medial degeneration was found in the aortic wall, suggesting toxic effect of GRF glue. The actuarial survival and root reoperation-free rates at 9 years were 73 and 80%, respectively. In conclusion, aortic root preservation with the proper use of GRF glue has long-term durability with very low adverse effect. PMID- 17416102 TI - [Adequate size of elephant trunk in the procedure of total arch replacement for acute aortic dissection]. AB - Total aortic arch replacement using the 'elephant trunk (ET)' procedure has commonly been applied to acute aortic dissection, but enlargement of a residual false lumen of the descending thoracic aorta sometimes occurs. We performed endovascular stent-grafting to close the entry as the second operation and obtained successful outcomes. From April 1997 to January 2004, we performed the modified ET procedure for acute aortic dissection in 29 patients and evaluated postoperative changes of the false lumen. In many cases of the residual false lumen, kinks and wrinkles were observed at the site of the ET grafts in the descending aorta. An adequate length of ET would be about 8 cm long to prevent kinking, and a diameter about 20 mm to prevent wrinkles. PMID- 17416103 TI - A brief history of tau: the evolving view of the microtubule-associated protein tau in neurodegenerative diseases. AB - The major historical milestones in tau-research are reviewed, with their implications for changing perspectives about the significance of tau-pathology in neurodegeneration. Abnormalities of tau-protein characterize the pathology of numerous neurodegenerative disorders, both sporadic and inherited. Over the years, opinions regarding the significance of tau in disease pathogenesis, particularly in Alzheimer's disease, have fluctuated. Early caution about the role of tau as a significant factor in neurodegenerative disease, especially Alzheimer's disease, has been superseded by acceptance of its key involvement in pathways which led to cell dysfunction and death. The discovery of familial "tauopathies", associated with tau-gene mutations, has confirmed that tau dysmetabolism can independently lead to neurodegeneration. Debate about the centrality of its role remains, but current evidence makes it difficult to ignore the importance of tau in many neurodegenerative diseases. By examining the evolution of research on tau, related to advances in technology and the emergence of new diseases, the future developments needed to resolve remaining issues in the tau-story may be discerned. PMID- 17416104 TI - Necrotizing neurosarcoid: three cases with varying presentations. AB - BACKGROUND: Neurosarcoid affects approximately 5% of patients with sarcoidosis. A significantly more rare entity, necrotizing sarcoidosis affecting the central nervous system, has been confirmed previously in only three case reports. This paper documents three additional cases of necrotizing neurosarcoid, involving a wide spectrum of central nervous system (CNS) locations. RESULTS: One patient presented to the emergency department after being found unresponsive. The second patient was referred due to hearing loss and the third patient sought care due to weakness and numbness of his left lower extremity. Locations of involvement were diverse and included diffuse leptomeningeal involvement, a cerebellopontine angle mass and a thoracic spinal cord lesion. All patients eventually underwent surgical biopsy, and histologic review of tissue samples revealed necrotizing granulomatous inflammation. Serum ACE levels were available for two of the patients and were within normal limits. Once the diagnosis of necrotizing neurosarcoid was confirmed, all patients were treated with systemic corticosteroid therapy; one patient was also treated with an immunosuppressive agent. CONCLUSIONS: Necrotizing neurosarcoid may occur more commonly than previously described and should be considered in the differential diagnosis of patients without systemic manifestations of sarcoidosis. PMID- 17416105 TI - Progressive multifocal leukoencephalopathy in a lymphoma patient with complete remission after treatment with cytostatics and rituximab: case report and review of the literature. AB - A 44-year-old woman presented with dysarthria, visual disturbances, ataxia and cognitive impairment. There was a rapid progression of her neurological disease, and she died 8 months later. She was previously treated for a low-grade follicular B-cell lymphoma; complete remission was achieved by conventional radiotherapy and chemotherapy, including rituximab. Two years later, the neurological symptoms and signs started. MRI revealed a cerebral demyelinating process. Serology was negative. Autopsy disclosed areas in cerebral white matter with grey discoloration. Microscopy revealed demyelination, oligodendroglial viral inclusions and gliosis with bizarre astrocytes. Polymerase chain reaction (PCR) was positive for JC virus. These findings were consistent with progressive multifocal leukoencephalopathy (PML). This is one of recent reports on PML occurring in a patient treated with the anti-20 monoclonal antibody rituximab. PMID- 17416106 TI - Three-dimensional analysis of pathological characteristics of a microaneurysm. AB - The etiology of Charcot-Bouchard microaneurysms (MAs), especially before rupture, has been unclear. CASE REPORT: A surgical specimen of an MA was removed from an acute intracerebral hemorrhage in a 69-year-old man. The MA was fixed with formalin and embedded in paraffin, and serial sections were cut and stained by the Azan and Elastica van Gieson methods and immunohistochemically. Three dimensional reconstructive analysis was performed. RESULTS AND DISCUSSION: The MA presented as local enlargement of an artery with a maximal diameter of 840 microm, but was not ruptured. The wall of the MA consisted of thickened, fine collagen fibrils, which were found to be similar to adventitia immunohistochemically, but with no internal elastic lamina or medial smooth muscle layer. The lumen was empty, with no mural thrombi, but the inner wall endothelial cells had expanded from the normal and/or residual arterial wall, which suggested that the MA had been present for a long period of time. Because the bare wall of the MA consisted of thickened collagen fibrils, the MA was not ruptured, but the possibility of rupture in the future was deemed high, with the risk of intracerebral hemorrhage. PMID- 17416107 TI - Effects of caffeic acid phenethyl ester on cerebral cortex: structural changes resulting from middle cerebral artery ischemia reperfusion. AB - Overproduction of free radicals is important in the pathogenesis of the cerebral damage induced by ischemia reperfusion. Caffeic acid phenethyl ester, an active component of propolis extract, exhibits antioxidant properties. The study was carried out in 16 male Wistar albino rats, divided into two groups: ischemia reperfusion and ischemia reperfusion with caffeic acid phenethyl ester. The middle cerebral artery was occluded for 60 min with an intraluminal suture, followed by 24-h reperfusion. In this study, widespread infarcted areas, red neurons (eosinophilic degeneration), pyknotic cells, vacuolization and neuroglial cell infiltration were observed in the cerebral cortex in the ischemia reperfusion group. In the caffeic acid phenethyl ester group, slightly infarcted areas were observed and neuroglial cell infiltration was not determined. Congestion of choroid plexus and pia mater was found more severe in the ischemia reperfusion group than in the caffeic acid phenethyl ester group. In the caffeic acid group, neuroglial cell activation was rare. Vacuolization, an indication of brain edema, was prevented by caffeic acid phenethyl ester. In the present study, we showed that pre-treatment with a single i.p. injection of caffeic acid phenethyl ester at 50 microM/kg dose reduced the structural changes. PMID- 17416108 TI - Intermittent administration of ceftazidime to burns patients: influence of glomerular filtration. AB - OBJECTIVE: The pharmacokinetics of ceftazidime, the antibiotic of choice for treating acute P. aeruginosa infections, may be modified in burns patients. The aim of this study was to identify the factors causing variations in the serum antibiotic concentrations in bums patients. METHODS: 30 patients with serious burns were randomly divided into two groups. Group 1 received a dose of ceftazidime of 2 x 3 g/24 hours. The second group received the same dose but divided into 6 administrations. Blood samples were taken at 24 (M1) and 48 hours (M2) after the start of treatment and the peak and trough serum concentrations of ceftazidime measured by HPLC. Depending on the results, frequency and/or dose was modified to obtain trough concentrations (Cmin) equal to 16 mg/l, i.e. 4 times the MIC. Either the same dose was maintained, but mostly divided up, or it was increased to 1 g x 8 administrations or it was decreased to 1 g x 4 or 1 g x 3. The serum concentrations of ceftazidime obtained were analyzed taking into account the characteristics of the burns patients (multivariate correlation). RESULTS: From the first sample (M1) Cmin was lower than the target concentration in 50% of the patients in Group 1 and 20% in Group 2. The modification of the dosing regimen put into place after the first analysis, led to the patients being further divided into four groups before the second blood sampling. Finally, 5 patients ended up in Group 1. In all patients and for all administration times, a negative correlation was found between Cmin and the creatinine clearance, calculated by using Cockcroft's formula. CONCLUSION: This study highlights the peculiarities of ceftazidime pharmacokinetics seen in burns patients with high interindividual variability. Based on Cmin monitoring and a predefined therapeutic range, dose adjustment was often required. Ceftazidime clearance is correlated with creatinine clearance (Cockcroft's formula), suggesting that this parameter could be used for a priori or a posteriori dose individualization. To respect the summary of the product characteristics (SPC) and reduce the variability in trough concentrations, the dose should be fractionated (1 g x 6) over a 24-hour period or even given as a continuous infusion. Trough concentrations must be evaluated to adapt the dosage regimen to attain target concentrations of 4 x the MIC. PMID- 17416109 TI - Isopropanolic black cohosh extract and recurrence-free survival after breast cancer. AB - OBJECTIVE: To investigate the influence of an isopropanolic Cimicifuga racemosa extract (iCR) on recurrence-free survival after breast cancer, including estrogen dependent tumors. METHODS: This pharmacoepidemiologic observational retrospective cohort study examined breast cancer patients treated at general, gynecological and internal facilities linked to a medical database in Germany. The main endpoint was disease-free survival following a diagnosis of breast cancer. The impact of treatment with iCR following diagnosis was analyzed by Cox-proportional hazards models, controlling for age and other confounders. RESULTS: Of 18,861 patients, a total of 1,102 had received an iCR therapy. The mean overall observation time was 3.6 years. Results showed that iCR was not associated with an increase in the risk of recurrence but associated with prolonged disease-free survival. After 2 years following initial diagnosis, 14% of the control group had developed a recurrence, while the iCR group reached this proportion after 6.5 years. The primary Cox regression model controlling for age, tamoxifen use and other confounders demonstrated a protractive effect of iCR on the rate of recurrence (hazard ratio 0.83, 95% confidence interval 0.69 0.99). This effect remained consistent throughout all variations of the statistical model, including subgroup analyses. TNM status was unknown but did not bias the iCR treatment decision as investigated separately. Hence, it was assumed to be equally distributed between treatment groups. Correlation analyses showed good internal and external validity of the database. CONCLUSION: An increase in the risk of breast cancer recurrence for women having had iCR treatment, compared to women not treated with iCR is unlikely. PMID- 17416110 TI - Intraoperative small-dose ketamine does not reduce pain or analgesic consumption during perioperative opioid analgesia in children after tonsillectomy. AB - OBJECTIVE: Ketamine inhibits the NMDA receptors via non-competitive antagonism, resulting in an antihyperalgesic effect achieved by doses of ketamine much smaller than are required for analgesia. The aim of this study was to determine the extent to which small-dose ketamine, when used in conjunction with remifentanil, has a morphine-sparing effect in the perioperative period. MATERIALS AND METHODS: In this randomized, double-blind, placebo-controlled prospective study, we enrolled 40 children undergoing tonsillectomy. Anesthetic care was standardized. Intraoperative analgesia was provided with remifentanil 0.5 microg x kg(-1) followed by an infusion of 0.25 microg x kg(-1) x min(-1). Group I (ketamine, n = 20) received a bolus dose of ketamine 0.5 mg x kg(-1) followed by a continuous infusion of 2 microg x kg(-1) x min(-1) before start of surgery. The infusion was stopped when surgery ended. Group II (placebo, n=20) received normal saline in the same manner. Pain was assessed postoperatively using the Children's Hospital Eastern Ontario Pain Scale (CHEOPS; range of scores 4 13), and total morphine consumption was recorded in the postanesthesia care unit (PACU). Patients were transferred to the ward and morphine was administered via a patient-controlled analgesia (PCA) device and analgesia was recorded using a visual analogue scale (VAS) (0 - 10). RESULTS: Intraoperative remifentanil consumption was not different between the ketamine group (0.29+/-0.09 microg x kg x min(-1) ) and the control group (0.24+/-0.07 microg x kg x min(-1)). There were no significant differences between CHEOPS scores and VAS score between the two groups. The total mean morphine consumption in the ward was not significantly different between the two groups: 376.5 +/-91.6 microg x kg(-1) with ketamine and 384.4+/-97.3 microg x kg(-1) with placebo. The time-to-first analgesic requirement was also similar in both groups. CONCLUSIONS: Small-dose ketamine did not decrease postoperative pain after tonsillectomy in children when added to a continuous intraoperative remifentanil infusion. PMID- 17416111 TI - Transgenic avian-derived recombinant human interferon-alpha2b (AVI-005) in healthy subjects: an open-label, single-dose, controlled study. AB - BACKGROUND/AIMS: This study characterized the safety and pharmacological properties of AVI-005, a novel glycosylated recombinant human interferon-alpha2b produced from the egg whites of chickens transfected with human cDNA. METHODS: 18 healthy volunteers received single subcutaneous rising doses (0.5, 1.66 or 5 million international units, MIU) of AVI-005. A randomized parallel comparator group of 10 subjects received 5 MIU of unglycosylated IFN-alpha2b (Intron A). The pharmacokinetic parameters t1/2, tmax, Cmax, AUC0-24h, Vd, and clearance were compared between AVI-005 and unglycosylated IFN-alpa2b. RESULTS: At equipotent doses, AVI-005 had a larger AUC0-24h than the control interferon. Pharmacodynamic markers ofneopterin and beta2-microglobulin for the two treatments were similar. These markers were increased by AVI-005 in a dose-dependent manner. Pharmacodynamic responses to treatment with AVI-005 were shown by the change in mRNA expression for interferon inducible protein kinase and 2'5'-oligoadenylate synthetase. Adverse events in the two groups were qualitatively and quantitatively similar. CONCLUSION: AVI-005 demonstrates biological activity and pharmaco-kinetic properties in humans that support further development. PMID- 17416112 TI - Antimicrobial use at a university hospital: appropriate or misused? A qualitative study. AB - OBJECTIVE: To evaluate the quality of antimicrobial drug use in a university hospital medical department (Department of Medicine, University Hospital Rijeka, Croatia) with 279 hospital-beds in wards containing patients from endocrinology, gastroenterology, hematology, clinical immunology, cardiology and coronary care unit, nephrology and pulmonology sections of the hospital. METHODS: The appropriateness of antimicrobial treatment for all in-patients in the Department of Medicine was assessed in a prospective, longitudinal survey carried out during a 21-week period using Kunin's criteria where Categories I and II indicate "appropriate therapy", Categories III and IV indicate major deficiency in the choice or use of antimicrobials. Category V indicates unjustified antimicrobial administration. RESULTS: During the study period, a total of 438 patients were treated with antimicrobials in the Department of Medicine. Of these, 159 (36%) received antimicrobials appropriately (Category I and II), 180 (41%) needed antimicrobials (Category III and IV) but they should have been prescribed differently. The main reason for inappropriate antimicrobial treatment was the wrong choice of antimicrobials (broad-spectrum where a narrow spectrum antibiotic would have been sufficient). In the case of 99 patients (23%) an indication for antimicrobial therapy did not exist (Category V). CONCLUSION: The main reason for suboptimal use of antimicrobials was the over-prescribing of broad-spectrum antimicrobials. This situation should be corrected e.g. by changes in the post graduate medical teaching program. PMID- 17416113 TI - A bioequivalence study of gliclazide based on quantification by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry. AB - OBJECTIVE: The aim of this study was to evaluate, in human volunteers, the performance of one gliclazide tablet formulation (gliclazide 80 mg tablet from EMS Industria Farmaceutica Ltda.) against two reference gliclazide tablet formulations (Diamicron 80 mg tablet from Servier do Brazil Ltda. and Diamicron 80 mg tablet from Servier (Ireland) Industries Limited). METHODS: The study had an open, randomized, three-period crossover design with a one-week washout interval between doses. The samples were obtained over a 48-h interval after each oral administration of gliclazide. The samples were extracted from plasma using diethylether : hexane (80 : 20, v/v) and the extracts were analyzed by high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-MS/ MS). Chromatography was performed isocratically using a Jones Chromatography Genesis C8 120A 4u. The method had a chromatographic run time of 2.5 min and a calibration curve of the range of 0.02- 10 microg x ml(-1) (r(2) > 0.9993). The limit of quantification was 0.02 microg x ml(-1). RESULTS: The geometric mean and 90% confidence intervals (CI) for the Gliclazide/Diamicron (Ireland) ratio were 588.68% (90% CI= 491.16, 705.58%) for AUClast, 423.50% (90% CI = 338.25, 530.23%) for AUCinf, and 1395.77% (90% CI= 1116.62, 1744.72%) for Cmax. The geometric mean and 90% confidence intervals (CI) for the Gliclazide/Diamicron (Brazil) ratio were 249.16% (90% CI = 207.96, 298.54%) for AUCiast, 249.16% (90% CI = 207.96 - 298.54%) for AUCinf, and 188.04% (90% CI - 151.72, 233.05%) for Cmax. CONCLUSION: Since the 90% CI for Cmax, AUClast and AUC(0-infinity) ratios were all outside the 125% interval proposed by the US Food and Drug Administration, we concluded that the gliclazide test formulation were not bioequivalent to either reference formulation. Interestingly, the pharmacokinetic parameters such as Cmax, AUClast of both reference formulations are compatible with neither the literature nor the profile of an immediate release formulation. In addition, both reference formulations were not bioequivalent in themselves, indicating significant differences in reference product formulation. PMID- 17416114 TI - Interaction between activated VD3 and Ca channel blockers in patients undergoing hemodialysis. PMID- 17416115 TI - Clinical impact of CYP2C19 polymorphism on the action of proton pump inhibitors: a review of a special problem. PMID- 17416117 TI - Conflict in the perioperative environment. PMID- 17416116 TI - Beverley McNeil writes on inadvertent hypothermia. PMID- 17416118 TI - I never could get the hang of Thursdays. PMID- 17416119 TI - Discharge criteria: 'just in case'. AB - According to the Association of Anaesthetists of Great Britain and Ireland (AAGBI 2002) the increasing complexity of surgical procedures has increased the need for improved monitoring of the postoperative patient. 'Recovery' of a patient has moved from a brief period of observation towards one of assessment and evaluation during the immediate postoperative phase. The purpose of this article is to explore current practice with a focus on the application and importance of discharge criteria. Critical analysis will highlight the responsibilities and accountability of the recovery practitioner (a registered nurse (RN) or operating department practitioner (ODP)) when discharging the patient. Conversely, the article will also explore the tensions in relation to effective application of the discharge criteria and the recovery practitioner's professional accountability. The term 'recovery practitioner' will be used throughout this paper to refer to those who are chiefly employed to care for patients in the recovery room, or more contemporarily referred to as the post anaesthetic care unit (PACU). PMID- 17416120 TI - United States operating room nurses: work environment perceptions. AB - Concerns about pending retirement of nurses working in the operating room (OR) are undeniable. The nurses' work environment and level of perceived support is part of the equation of why a nurse may choose to retire or stay in the workforce. This study compares nurses' perceptions of the work environment between OR nurses and nurses who work in other type units; and compares the work environment perceptions of OR nurses between institutions of two sizes (<300 beds and >300 beds). Findings include: OR nurses have better perceptions of their work environment than nurses in other type units and OR nurses in smaller hospitals are more satisfied with their work environment, workload and perceptions of organisational support. Implications for OR work environments are discussed. PMID- 17416121 TI - Preoperative hair removal: a systematic review. AB - Preparing patients for surgery has traditionally included the routine removal of body hair from the intended surgical wound site. However, there are studies which claim that preoperative hair removal is deleterious to patients, perhaps by causing surgical site infections (SSIs), and should not be carried out. The objective of this review was to determine if routine preoperative hair removal results in fewer SSIs than not removing hair. Eleven randomised controlled trials were included in this review. There is insufficient evidence to state whether removing hair impacts on surgical site infection or when is the best time to remove hair. However, if it is necessary to remove hair then both clipping and depilatory creams results in fewer SSIs than shaving using a razor. PMID- 17416122 TI - The Smith-Petersen trifin nail. PMID- 17416123 TI - The effect of anterior versus posterior glide joint mobilization on external rotation range of motion in patients with shoulder adhesive capsulitis. AB - STUDY DESIGN: Randomized clinical trial. OBJECTIVE: To compare the effectiveness of anterior versus posterior glide mobilization techniques for improving shoulder external rotation range of motion (ROM) in patients with adhesive capsulitis. BACKGROUND: Physical therapists use joint mobilization techniques to treat motion impairments in patients with adhesive capsulitis. However, opinions of the value of anterior versus posterior mobilization procedures to improve external rotation ROM differ. METHODS AND MEASURES: Twenty consecutive subjects with a primary diagnosis of shoulder adhesive capsulitis and exhibiting a specific external rotation ROM deficit were randomly assigned to 1 of 2 treatment groups. All subjects received 6 therapy sessions consisting of application of therapeutic ultrasound, joint mobilization, and upper-body ergometer exercise. Treatment differed between groups in the direction of the mobilization technique performed. Shoulder external rotation ROM measured initially and after each treatment session was compared within and between groups and analyzed using a 2-way ANOVA, followed by paired and independent t tests. RESULTS: There was no significant difference in shoulder external rotation ROM between groups prior to initiating the treatment program. A significant difference between groups (P = .001) was present by the third treatment. The individuals in the anterior mobilization group had a mean improvement in external rotation ROM of 3.0 degrees (SD, 10.8 degrees; P = .40), whereas the individuals in the posterior mobilization group had a mean improvement of 31.3 degrees (SD, 7.4 degrees; P < .001). CONCLUSIONS: A posteriorly directed joint mobilization technique was more effective than an anteriorly directed mobilization technique for improving external rotation ROM in subjects with adhesive capsulitis. Both groups had a significant decrease in pain. PMID- 17416124 TI - Efficacy of a C1-C2 self-sustained natural apophyseal glide (SNAG) in the management of cervicogenic headache. AB - STUDY DESIGN: Randomized, double-blind, placebo controlled trial. OBJECTIVES: To determine the effect of a C1-C2 self-sustained natural apophyseal glide (SNAG) on cervicogenic headache. BACKGROUND: Cervicogenic headache is a common condition causing significant disability. Recent studies have shown a high incidence of C1 C2 dysfunction, evaluated by the flexion-rotation test (FRT), in subjects with cervicogenic headache. To manage this dysfunction, Mulligan has described a C1-C2 self-SNAG, though no studies have investigated the efficacy of this intervention approach. METHODS: A sample of 32 subjects (mean _ SD age, 36 +/- 3 years) with cervicogenic headache and FRT limitation were randomized into a C1-C2 self-SNAG or placebo group. After an initial instruction and practice visit in the clinic, interventions consisted of exercises applied independently by the subject twice daily at home on a continual basis. FRT range was measured twice, before and immediately after the instruction and practice visit. Headache symptoms were determined by a headache index over time, assessed by questionnaire preintervention, at 4 weeks postintervention, and at 12 months postintervention. RESULTS: No differences were found in baseline measures between groups. Immediately after the initial instruction and practice visit performed with the supervision of the therapist, FRT range increased by 15 degrees (SD, 9) for the C1-C2 self-SNAG group (P < .001), which was significantly more than 5 degrees (SD, 5) for the placebo intervention (P < .001). There was also a significant interaction for the variable headache index between group and time (P < .001), indicating that group difference was dependent on time. There was no difference in headache index scores at baseline between groups. Headache index scores were substantially less in the C1-C2 self-SNAG group (mean +/- SD points at 4 weeks, 31 +/- 9; mean +/- SD points at 12 months, 24 +/- 9) compared to the placebo group (mean +/- SD points at 4 weeks, 51 +/- 15; mean +/- SD points at 12 months, 44 +/- 13) at 4 weeks (P < .001) and 12 months (P < .001), with an overall (+/ SD) reduction of 54% (+/-17%) for the individuals in the C1-C2 self-SNAG group. CONCLUSIONS: These results provide evidence for the efficacy of the C1-C2 self SNAG technique in the management of individuals with cervicogenic headache. PMID- 17416125 TI - A randomized controlled comparison of stretching procedures for posterior shoulder tightness. AB - STUDY DESIGN: Randomized controlled trial, OBJECTIVES: To compare changes in shoulder internal rotation range of motion (ROM), for 2 stretching exercises, the "cross-body stretch" and the "sleeper stretch," in individuals with posterior shoulder tightness. BACKGROUND: Recently, some authors have expressed the belief that the sleeper stretch is better than the cross-body stretch to address glenohumeral posterior tightness because the scapula is stabilized. METHODS: Fifty-four asymptomatic subjects (20 males, 34 females) participated in the study. The control group (n=24) consisted of subjects with a between-shoulder difference in internal rotation ROM of less than 10 degrees, whereas those subjects with more than a 10 degrees difference were randomly assigned to 1 of 2 intervention groups, the sleeper stretch group (n=15) or the cross-body stretch group (n=15). Shoulder internal rotation ROM, with the arm abducted to 90 degrees and scapula motion prevented, was measured before and after a 4-week intervention period. Subjects in the control group were asked not to engage in any new stretching activities, while subjects in the 2 stretching groups were asked to perform stretching exercises on the more limited side only, once daily for 5 repetitions, holding each stretch for 30 seconds. RESULTS: The improvements in internal rotation ROM for the subjects in the cross-body stretch group (mean +/- SD, 20.0 degrees +/- 12.9 degrees) were significantly greater than for the subjects in the control group (5.9 degrees +/- 9.4 degrees, P = .009). The gains in the sleeper stretch group (12.4 degrees +/- 10.4 degrees) were not significant compared to those of the control group (P = .586) and those of the cross-body stretch group (P = .148). CONCLUSIONS: The cross-body stretch in individuals with limited shoulder internal rotation ROM appears to be more effective than no stretching in controls without internal rotation asymmetry to improve shoulder internal rotation ROM. While the improvement in internal rotation from the cross body stretch was greater than for the sleeper stretch and of a magnitude that could be clinically significant, the small sample size likely precluded statistical significance between groups. PMID- 17416126 TI - Sex differences in pain drawing area for individuals with chronic musculoskeletal pain. AB - STUDY DESIGN: Cross-sectional. OBJECTIVES: To (1) determine the association between pain severity and pain drawing area for men and women; (2) determine if sex differences exist in pain severity or pain drawing area; (3) determine the relative influence of pain severity, anatomical location of pain, personality, and psychological coping factors on pain drawing area for men and women. BACKGROUND: Pain drawings have been postulated to assist in clinical decision making regarding classification and treatment of musculoskeletal pain. Prior studies have been ambiguous on this topic, possibly because they have not considered if sex differences exist for pain drawing area. METHODS AND MEASURES: One hundred twenty-six subjects referred to a multidisciplinary chronic pain clinic with chronic musculoskeletal pain were included in this study. Subjects completed a pain drawing, the Multidimensional Pain Inventory (MPI), the Coping Strategies Questionnaire (CSQ), and the Minnesota Multiphasic Personality Inventory (MMPI-2). Pearson correlations investigated the associations of pain severity and pain drawing area, independent t tests investigated sex differences in pain severity and pain drawing area, and multiple regression investigated factors that influenced pain drawing area. RESULTS: P a in severity w as positively correlated with pain drawing area for men (r = 0.38, P = .003) and women (r = 0.23, P = .052), accounting for approximately 14% and 5% of the total variance, respectively. There was no significant sex difference in pain severity ratings, but women reported a significantly larger area of symptoms on the pain drawings (effect size, 0.61; P = .002). The sex difference in pain drawing area was consistent across different anatomical locations of pain. In women, the final regression model accounted for 39% (P < .001) of the variance in pain drawing area, with anatomical location of pain (beta = .42, P < .001) and hypochondriasis (beta = .31, P = .005) as the only unique predictors in the final model. In men, the regression model accounted for 27% (P = .003) of the variance in pain drawing area, with pain severity (beta = .32, P = .021) and a coping style of ignoring pain (beta = -.32, P = .018) as the only unique predictors in the final model. CONCLUSIONS: Women had larger pain drawing area and this area was significantly associated with anatomical location of pain and hypochondriasis. Men had smaller pain drawing area and this area was associated with pain severity and a coping style of ignoring pain. These findings suggest that clinicians interpreting pain diagram area should consider the sex of the individual. PMID- 17416127 TI - Investigation of clinician agreement in evaluating movement quality during unilateral lower extremity functional tasks: a comparison of 2 rating methods. AB - STUDY DESIGN: Nonexperimental. OBJECTIVES: To determine interrater and intrarater agreement for 2 methods of evaluating movement quality during 2 lower extremity functional tasks, and to descriptively compare levels of agreement between the 2 methods. BACKGROUND: Clinicians typically use observational analysis to evaluate movement quality during functional tasks, but the extent of agreement is unknown. METHODS AND MEASURES: Twenty-five uninjured subjects performed 3 trials of unilateral squat and lateral step-down tasks. Three clinicians evaluated the trunk, pelvis, and hips for coronal plane and transverse plane movement deviations. Two rating methods were used: assessment of the entire movement ("overall method") and rating each segment individually ("specific method"). Movement deviation severity was rated using basic clinical guidelines and ratings were repeated from videotape. Percent agreement and weighted kappa coefficients were calculated between rater pairs and rating sessions. Generalized kappa coefficients were calculated across raters. RESULTS: Interrater and intrarater percent agreement were higher using the overall method. Interrater weighted kappa coefficients were similar between rating methods (overall method, 0-0.55; specific method, 0.23-0.53). Intrarater weighted kappa coefficients were higher for the specific method (0.38-0.68) compared to the overall method (0.13-0.50). Generalized kappa coefficients were also higher for specific method compared to the overall method (unilateral squat, 0.19 and 0.01, respectively; lateral step down, 0.22 and 0.18, respectively) and 95% confidence intervals remained above zero. CONCLUSIONS: Rating movement at body segments appears to result in agreement among raters that is better than chance. Neither rating method produced high agreement, indicating a need to develop more explicit criteria for rating movement deviation severity. PMID- 17416128 TI - An acute bout of static stretching does not affect maximal eccentric isokinetic peak torque, the joint angle at peak torque, mean power, electromyography, or mechanomyography. AB - STUDY DESIGN: Repeated-measures experimental design. OBJECTIVE: To examine the acute effects of static stretching on peak torque, the joint angle at peak torque, mean power output, and electromyographic and mechanomyographic amplitudes and mean power frequency of the vastus lateralis and rectus femoris muscles during maximal eccentric isokinetic muscle actions. BACKGROUND: A bout of static stretching may impair muscle strength during isometric and concentric muscle actions, but it is unclear how static stretching may affect eccentric force production. METHODS AND MEASURES: Fifteen men (mean +/- SD age, 23.4 +/- 2.4 years) performed maximal eccentric isokinetic muscle actions of the dominant and nondominant knee extensor muscles at 60 degrees x s(-1) and 180 degrees x s(-1) on an isokinetic dynamometer, while electromyographic and mechanomyographic amplitudes (root-mean-square) and mean power frequency were calculated for the vastus lateralis and rectus femoris muscles. Peak torque (Nm), the joint angle at peak torque (degrees), and mean power output (W) values were recorded by the dynamometer. Subsequently, the dominant lower extremity knee extensors underwent static stretching exercises, then the assessments were repeated. RESULTS: There were no stretching-related changes in peak torque, the joint angle at peak torque, mean power output, electromyographic or mechanomyographic amplitude, or mean power frequency (P > .05). However, there were expected velocity-related, limb-related, and muscle-related differences (P < or = .05) that were unrelated to the stretching intervention. CONCLUSION: These results suggest that static stretching does not affect maximal eccentric isokinetic torque or power production, nor does it change muscle activation. PMID- 17416129 TI - Insidious onset of shoulder girdle weakness. AB - STUDY DESIGN: Resident's case problem. BACKGROUND: An 18-year-old man presented to physical therapy 3 days after insidious onset of painless left shoulder girdle weakness. DIAGNOSIS: Decreased light touch sensation was noted on the lateral left shoulder. In addition, weakness was present with shoulder abduction, flexion, external rotation, and internal rotation. Results of magnetic resonance imaging and radiography of the cervical spine, brachial plexus, and left shoulder were normal, Electromyography and nerve conduction velocity study findings were consistent with axillary nerve palsy. The results of the physical examination and diagnostic studies were most consistent with axillary nerve mononeuropathy, probably caused by traction or pressure due to wearing a pack while hiking or firing a weapon. DISCUSSION: With sling protection, limitation of physical activity, and gradual return to progressive resistance exercises, the patient had full return of strength and function 2 1/2 months after onset of symptoms. The differential diagnosis for shoulder girdle weakness should be well understood by physical therapists. This knowledge will help the therapist promptly identify the cause of shoulder girdle weakness and initiate appropriate treatment. If the condition requires further evaluation or treatment by another healthcare provider, prompt identification of pathology will allow appropriate timely referral. PMID- 17416130 TI - [Cognitive enrichment in farm animals--the impact of social rank and social environment on learning behaviour of dwarf goats]. AB - The influence of social rank and social environment on visual discrimination learning of small groups of Nigerian dwarf goats (Capra hircus, n = 79) was studied using a computer-controlled learning device integrated in the animals' home pen. The experiment was divided into three sections (LE1, LE1 u, LE2; each 14d). In LE1 the goats learned a discrimination task in a socially stable environment. In LE1u animals were mixed and relocated to another pen and given the same task as in LE1. In LE2 the animals were mixed and relocated again and given a new discrimination task. We used drinking water as a primary reinforcer. The rank category of the goats were analysed as alpha, omega or middle ranking for each section of the experiment. The rank category had an influence on daily learning success (percentage of successful trials per day) only in LE1 u. Daily learning success decreased after mixing and relocation of the animals in LE1 u and LE2 compared to LE1. That resulted in an undersupply of drinking water on the first day of both these tasks. We discuss social stress induced by agonistic interactions after mixing as a reason for that decline. The absolute learning performance (trials to reach the learning criterion) of the omega animals was lower in LE2 compared to the other rank categories. Furthermore, their absolute learning performance was lower in LE2 compared to LE1. For future application of similar automated learning devices in animal husbandry, we recommend against the combination of management routines like mixing and relocation with changes in the learning task because of the negative effects on learning performance, particularly of the omega animals. PMID- 17416131 TI - Expert opinion based modelling of the risk of human infection with H5N1 through the consumption of poultry meat in Germany. AB - According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model. The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation. The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply. The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia. PMID- 17416132 TI - Rotaviruses: diversity and zoonotic potential--a brief review. AB - Rotaviruses, a genus within the family Reoviridae, are among the most important etiological agents of severe diarrhoeal illness in humans and animals worldwide. Their genome, consisting of 11 segments of double-stranded RNA, is characterized by genetic variability including (i) point mutations, (ii) genomic reassortment, and (iii) genome rearrangements, thus leading to the considerable diversity of rotaviruses. Animal rotaviruses are regarded as a potential reservoir for genetic exchange with human rotaviruses. There is now increasing evidence that animal rotaviruses can infect humans, either by direct transmission of the virus or by contributing one or several genes to reassortants with essentially a human strain genetic background. As mixed infections are a prerequisite for reassortment events, cosurveillance of animal and human rotavirus strains will be vital to gain a better understanding of the relationships between cocirculating viruses, as well as assessing any relevant vaccination programs. PMID- 17416133 TI - [Detection of DNA of the finch polyomavirus in diseases of various types of birds in the order Passeriformes]. AB - Between 2000 and 2004 a disease occurred in an aviary in Germany affecting various bird species belonging to the order Passeriformes including Collared Grosbeaks (Mycerobas affinis), Eurasian Bullfinches (Pyrrhula pyrrhula griseiventris), Brown Bullfinches (Pyrrhula nipalensis), Grey-headed bullfinches (Pyrrhula erythaca) and Yellow-bellied Tits (Periparus venustulus). The major clinical signs included increased mortality of fledglings and young birds, as well as feather disorders and feather loss in adult birds. In addition, adult Eurasian Bullfinches showed in one year a disease course, in which the major symptom was inflammation of the skin beginning on the basis of the beak and spreading over the head occurring a few days before death. Bacteriological and parasitological investigations did not reveal any consistent findings. Using a newly developed polymerase chain reaction protocol, DNA of the recently discovered finch polyomavirus (FPyV) was demonstrated in several affected birds. Because of the consistent detection of FPyV-DNA and the similarity of the symptoms with those observed during infection with the closely related avian polyomavirus in other bird species, an etiological role of FPyV in the observed disease is assumed. PMID- 17416134 TI - [Bacteriological and virological status in upper respiratory tract infections of cats (cat common cold complex)]. AB - Between October 2002 and January 2005,460 bacteriological samples from cats with an acute upper respiratory tract infection were analysed in clinical field studies in two accredited laboratories in Germany. Oropharyngeal swabs were taken from these cats and sent to the laboratories for routine diagnostics. In the swab samples of 460 cats 382 bacteria strains were isolated. The following bacteria were isolated most frequently: Pasteurella spp. (32.5 %), Staphylococcus spp. (18.5 %), Escherichia coli (17.0 %), Streptococcus spp. (9.1 %), Pseudomonas spp. (6.9 %) and Klebsiella spp. (3.0 %). Bordetella bronchiseptica was found in 0.4 % of the animals To evaluate possible regional and time influences, the animals were split into three populations: 1: Germany, laboratory A; 2: Germany, laboratory B; 3: France and Belgium, laboratory B. In population 1 an 2 Pasteurella spp. were found most frequently with 42.2 % and 36.5 %, respectively. The second most frequently isolated bacterial species were Staphylococcus spp. with 14.1 % and 21.4 % and E. coli with 13.6 % and 17.5 % respectively. In population 3 Staphylococcus spp., E. coli (20 % each) and Pasteurella spp. (18.5 %) were isolated at almost the same frequency. Virological parameter were additionally analysed in 328 cats (population 2 and 3). Serum samples were analysed for antibodies specific for Feline Calicivirus (FCV) and Feline Immunodeficiency Virus (FIV) and for Feline Leukaemia Virus (FeLV) antigen. Oropharyngeal swabs were analysed for Feline Herpesvirus (FHV) by using PCR. Calicivirus-specific antibodies were found in 99.6 % of the cats of population 2 and in 100 % of the animals in population 3. Herpesvirus was detected in 15.3 % and 23.3 % of the cats, respectively. FeLV-Antigen was found in 0.4 % of the animals in population 2 and in 10.1 % of the cats in population 3, while FIV antibodies were identified in 8.7 % of the animals of population 2 and in 6.1 % of the cats of population 3. In total FHV was found in 19.3 % and FCV-specific antibodies in 99.7 % of the animals. 5.3 % of the cats carried FeLV-Antigen, and 7.4 % FIV-specific antibodies. The results of the bacteriological analysis as well as the results of the virological examination confirm previously published data. In this study Pasteurella spp. were most frequently isolated (32.5 %). PMID- 17416135 TI - [Detection of rhodococcus equi by microbiological culture and by polymerase chain reaction in samples of tracheobronchial secretions of foals]. AB - The goal of the present study was to investigate whether new PCR-methods would improve diagnostic of R. equi. In a first step, sensitivity and specificity of the PCR-methods in respect to the"gold standard" microbiological culture were determined. Secondly, sensitivity and specificity of both microbiological methods were evaluated in respect to the clinical diagnosis. The tracheobronchial secretions of 48 foals with pulmonary abscesses and of 37 healthy foals were evaluated by bacteriological culture as well as by four PCR-methods: aceA-, ideR , vapA- and VP-PCR. In respect to the"gold standard" microbiological culture, the sensitivity of most PCR methods lay between 63.9 and 69.4 % except the vapA-PCR (27.8 %). The specificity of all PCR methods in this comparison was between 98 to 100 %. In this analysis, clinical diagnosis had a low sensitivity (66.7 %) and a low specificity (51.0 %). In respect to the clinical diagnosis, microbiological culture sensitivity was 50.0 % and specificity 67.7 9%. In this analysis, sensitivity rates of aceA-, ideR and VP-PCR methods lay between 33.3 and 37.5 %, sensitivity of the vapA-PCR was lower (10.4 %). The specificity of all PCR methods ranged from 78.4 to 86.5 %. In conclusion, these results show that the diagnostic potential of the microbiological methods"Culture" and "PCR" is different and that for the diagnosis of R. equi-pneumonia in foals the combination of microbiological culture with PCR should be used for examination of samples of the airways of foals. PMID- 17416136 TI - Metabolic and clinical traits in horses undergoing feed deprivation for elective orthopaedic surgery. AB - The objective of this study was to investigate some metabolic and clinical effects of feed deprivation in horses that were submitted for orthopaedic surgery. The effects of preoperative feed restriction were investigated in 20 horses submitted for elective orthopaedic surgery. The patients were fasted from 12 hours before until 4 hours after surgery. Serum free amino acids, glucose,free fatty acids (FFA), white blood cell counts, creatine kinase (CK) and aspartate aminotransferase (AST) were determined 24 hours before surgery, 2 hours after the end of anaesthesia and 24 and 72 hours after surgery. Besides, abdominal sounds, appetite, faecal quality and body temperature were examined. Serum free amino acids did not react homogenously, concentrations were partly increasing or decreasing. Plasma glucose and FFA increased after surgery and returned to their preoperative levels 72 hours after surgery. A significant rise of the segmented granulocytes occurred 24 hours after surgery, all other parameters of the leukogram did not exceed the physiological range. AST reached its highest activity 24 hours after surgery, whereas CK activities were highest at 2 hours after surgery. Abdominal sounds were significantly reduced until 24 hours after surgery, however, appetite was not depressed. Faecal quality was physiological after surgery. Mean body temperature stayed within the physiological range. In conclusion, a relatively short perioperative fasting period had significant effects on the metabolic traits in horses, however the effects on physiological functions were minor. The consequences of major surgical procedures need to be addressed in future studies. PMID- 17416137 TI - [Biodegradation of estrogens in stream water]. AB - Estrogens of human and animal origin that reach the aquatic environment may enter human or animal organism and act as endocrine disruptors.To investigate the persistency of estrogens in laboratory experiments, estrone respectively 17beta estradiol were added to stream water sampled from river Spree in Berlin. The concentration of estrone and 17beta-estradiol was quantified using enzyme-immuno assay. The estrone concentration decreased to less than 5 % of the starting concentration at storage temperature of 5 degrees C within 56 days and at storage temperature of 20 degrees C within 14 days. If the estrone were added to autoclaved stream water, no biodegradation was observed. Biodegradation was enhanced when activated sludge was added. Escherichia coli, Pseudomonas fluorenscens and Aeromonas hydrophila in monoculture did not degrade estrone in autoclaved stream water. The concentrations of 17beta-estradiol and estrone decreased similarly. The logistic function proved to be suitable to describe the course of time for the decrease of concentration. PMID- 17416138 TI - [Qualification of the Stewart variables for the assessment of the acid-base status in healthy dogs and dogs with different diseases]. AB - Peter Stewart criticized the traditional theory of the acid-base status by Henderson-Hasselbalch as too simple and incomplete. He developed a new model with 3 independent variables: (1) pCO2, (2) SID (strong ion difference) and (3) Atot (Acid total). In healthy and ill dogs the diagnostic usefulness of both acid-base models were compared. This study included n=58 healthy dogs and 3 clinical cases of sick dogs. The age of the healthy dogs was 5.0 (2.0-7.0) years (= median (1. 3. quartil)). The 3 clinical cases included (1) a dog with septic shock, (2) with acute renal insufficiency, and (3) with hypovolaemic shock due to gastric torsion. Venous blood was taken of all dogs and the acid-base parameters were determined within < or =30 minutes. Electrolytes and albumin were determined in blood serum and used for calculation of the Stewart variables. Limits of reference intervals (x+/-1.96 - s) were determined for the healthy dogs yielding pCO2 = 3.6-6.5 kPa, [SID3] = 33.1-50.9 mmol/l resp. [SID4] = 31.8-49.6 mmol/l and [Al = 8.5-13.1 mmol/l. In Case 1 the Henderson-Hasselbalch parameters demonstrated the presence of a strong metabolic acidosis with mild respiratory influence (pH, [HCO3-], [BE] and PCO2 at upper range of normal). Analysis of the Stewart variables [SID3] resp. [SID4] revealed an electrolyte imbalance with [Cl ] and [lactate-] as the reason for metabolic acidosis. Case 2 showed a metabolic acidosis with respiratory compensation (pH, [HCO3-], [BE] and PCO2). Analysis of the Stewart variables with [SID3] resp. [SID4 caused by [K+], [Na+] and [lactate ]demonstrated the acidotic metabolism due to a renal malfunction. Case 3 had a metabolic acidosis (pH-value in the lower range) caused by electrolyte imbalances ([SID4]. The Stewart variables allow a better understanding of the causes of acid base-disturbances in animals with implications for successful therapy via infusion. PMID- 17416139 TI - [Resection of the distal/proximal interphalangeal joint and digit amputation in 21 breeding bulls--indications, clinical findings and longterm outcome]. AB - Case records of 21 breeding bulls suffering from advanced septic claw disorders and treated with resection of the distal interphalangeal (= DIP) joint (n = 14), resection of the proximal interphalangeal joint (n = 1) and digit amputation (n = 6) were evaluated retrospectively (1992-2005) regarding the following parameters: breed, age, body mass, pre-treatment, localisation of disease, lameness degree, diagnosis, surgical method, outcome, postoperative survival time and return to breeding use. The mean age of the bulls was 3.6+/-1.7 years and the mean body mass 840.7 + 200.8 kg; range: 500-1200 kg), Surgery was carried out to treat the following conditions: white line infection (n = 9), sole ulcers (n = 5) with osteolysis of the distal sesamoid bone, the tuberculum flexorium, infection of the deep digital flexor tendon at insertion and the DIP-joint; osteomyelitis of the distal and middle phalanx with septic arthritis of the DIP-joint following deep penetrating wounds (n = 3); open fracture of the coffin bone with severe osteolysis/osteomyelitis and septic fibrino-purulent arthritis of the DIP-joint (n = 2); purulent arthritis of the proximal interphalangeal joint after a penetrating wound (n = 1) and a severe osteolysis/osteomyelitis of the coffin bone originating from a toe ulcer (n = 1). Three bulls were culled within 2-12 weeks after surgery due to postsurgical complications. In three bulls which were treated successfully six years ago, long-term follow-up information could not be obtained. Complete recovery was considered in 15 bulls. They returned to full breeding use three to four months after surgery, with a mean postoperative survival time of 24.1 months:21.2 months after joint resection and 27.2 months after amputation. At the time of this survey three bulls were still alive with 4,7 and 84 months after surgery, respectively. Both surgical methods were successful in breeding bulls, the resection of the distal interphalangeal joint is recommended as first option. PMID- 17416140 TI - Effect of different claw trimming methods on the pressure distribution under the bovine claw--an in vitro study. AB - This survey focusses on the effects of various claw trimming methods as well as the effect of the different resulting claw shapes on the pressure distribution under the sole. 64 bovine claws were trimmed according to 3 different trimming methods, the functional claw trimming method by Toussaint Raven and two other specially modified methods. The alternative methods resulted on the one hand in long and acute-angled claws, on the other hand in a very steep claw profile. The limb samples were attached to the hydraulic plunger of a material testing machine and pressed onto a pressure distribution plate with a predefined load. The pressure distribution pattern was recorded before and after trimming. An obvious stress concentration could be observed in the bulbar region of the outer claws. All of the applied claw trimming methods induced a redistribution of load onto the inner claws while relieving the bulbar area of the outer claws. The inner claws were the main reason for the enlarged floor contact area caused by the trimming techniques. The results show that all techniques led to an improvement of stress conditions. In some cases, methods 2 and 3 were slightly superior in reducing maximum pressure and enlarging floor contact area. PMID- 17416141 TI - Approved: The Joint Commission's hospital performance measurement implementation plan through 2008. PMID- 17416142 TI - Approved: revisions to 2007 National Patient Safety Goals and Universal Protocol. PMID- 17416143 TI - Correction: Standard LD.2.140, Element of Performance 9, for laboratory. PMID- 17416144 TI - Correction: Strategic surveillance system information. PMID- 17416145 TI - Revision: Standard HR.4.20, Element of Performance 16, for ambulatory care. PMID- 17416146 TI - How long should a course of rehabilitation be? PMID- 17416147 TI - Four weeks' intensive rehabilitation generates significant health effects in COPD patients. AB - Changes in health according to World Health Organization's International Classification of Functioning, Disability and Health (ICF) after four weeks of pulmonary rehabilitation (PR) were investigated. Gender differences in the response to PR, and the correlation between improvements in the two components of ICF (Body functions and Activities and Participation) were examined. Twenty-two men and 18 women with chronic obstructive pulmonary disease in Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage II-IV attended in-patient, multidisciplinary PR consisting of endurance training four to five times/week at 70% of peak work rate (WRpeak), resistance training three to four times/week at 72% of 15 repetitions maximum, educational sessions and individual counselling. The results were compared to those of 20 Chronic Obstructive Pulmonary Disease (COPD) patients included after the same criteria and investigated while waiting for admission to PR. In the rehabilitation group, we found significant improvements in health related quality of life (HRQoL) (-7 units, St. George's Respiratory Questionnaire), arm (6%) and leg (15%) maximal voluntary contraction, peak oxygen uptake (6%), WRpeak (60%) and treadmill endurance time (93%). At iso WR, ventilation and dyspnoea were significantly lower, but inspiratory capacity remained unchanged. Improvements in HRQoL correlated with increases in peak ventilation, but not in muscle strength or exercise capacity. Men improved their six-minute walking distance significantly in contrast to women. Clinically important improvements in HRQoL were found in two out of three of the men, and one out of three of the women. Four weeks of intensive PR generated significant health effects comparable to longer lasting programmes. Changes in exercise capacity and muscle strength were not related to improvements in HRQoL. The gender differences in the response to PR deserve attention in future studies. PMID- 17416148 TI - Assessment of sleep studies in myotonic dystrophy. AB - Myotonic dystrophy (MD) is the commonest adult muscular dystrophy and is associated with respiratory muscle weakness. The role of screening sleep studies is unclear in MD. We prospectively evaluated polysomnography/overnight oximetry in a group of MD patients and related this to the daytime respiratory function in an attempt to evaluate the usefulness of screening sleep studies. Twenty-five patients with type I MD [15 males; mean age (SD) 40.0 (10.9) years] who had at least one symptom suggestive of nocturnal hypoventilation were included in the study. We performed spirometry, maximal inspiratory and expiratory mouth pressures, sniff nasal inspiratory pressure, arterial blood gases and polysomnography or overnight oximetry. Excessive tiredness and sleepiness were the most common presenting symptoms. Prevalence of sleep related breathing disorder (SRBD) was 36%. FVC was found to be normal in 33% of subjects with significant SRBD. Mouth pressures were reduced more than FVC, even in patients with normal overnight oxygen saturation. Of all the daytime measures, FVC correlated best with arterial carbon dioxide tension (r = -0.7). Sleep studies were useful to identify a small group of myotonic dystrophy patients (12%, three out of 25 in our series) with SRBD that would have been missed with routine daytime assessments. Targeted sleep monitoring in patients who are older, with multiple symptoms suggestive of SRBD, especially if they are overweight seems to be the best way to utilize the existing resources. Home unattended oximetry was well tolerated and offers a practical screening tool in this challenging patient group where excess daytime sleepiness is often due to causes other than SRBD. PMID- 17416149 TI - Pulmonary function tests in children with beta-thalassemia major. AB - Lung function abnormality is a known complication of thalassemia, but the results of studies in pulmonary function have been inconsistent. This study was conducted to describe the type of lung impairment in thalassemic children. Pulmonary function tests were conducted in 40 children with beta-thalassemia major, 23 males and 17 females. Tests included spirometry, total lung capacity (TLC), single breath diffusing capacity of the lung for carbon monoxide (DL(CO)) and arterial blood gases. Serum ferritin level was measured in all children to study its relationship to lung function impairment. A predominantly restrictive pattern was seen in 14 patients (35%). These patients had a significant reduction in RV, FVC, TLC and PEF with an FEV1/FVC ratio of more than 75%. Obstructive airway disease was found in six patients (15%), with an FEV1/FVC ratio less than 75%, increased RV and reduced FEF(25%-75%). Impairment of diffusion was found in 10 patients (25%), with DL(CO) reduced to less than 80% of the predicted value. Arterial blood gases results showed that no patient was hypoxic. No correlation was found between the severity of restrictive or obstructive disease and the serum ferritin level. There was a significant linear correlation between age and serum ferritin level (P < 0.019). Patients with thalassemia have a predominantly restrictive lung dysfunction pattern. This may be due to pulmonary parenchymal pathology, although the reason for the obstructive pattern seen in a small proportion of patients remains obscure. PMID- 17416150 TI - A randomized evaluation of the acute efficacy, acceptability and tolerability of flutter and active cycle of breathing with and without postural drainage in non cystic fibrosis bronchiectasis. AB - Airway clearance is integral to the management of bronchiectasis, yet there is no evidence as to the optimal modality. The aim of this randomized prospective study was to evaluate the acute efficacy, acceptability and tolerability of three airway clearance techniques in non-cystic fibrosis (non-CF) bronchiectasis. Flutter, active cycle of breathing technique (ACBT) and ACBT with postural drainage (ACBT-PD) were evaluated in random order over a week in 36 patients (mean age 62 years, range 33-83), with stable non-CF bronchiectasis. Total sputum wet weight for ACBT-PD was twice that of either ACBT alone or Flutter. No objective difference in treatment duration was noted. All three techniques were well accepted and tolerated. Patient preference was 16 (44%) for Flutter, eight (22%) ACBT and 12 (33%) for ACBT-PD. Patient demography, factors such as upper airways or reflux symptoms, previous use or acute efficacy did not predict preference. This is the first randomized systematic evaluation of acute efficacy, acceptability and tolerability of Flutter, ACBT and ACBT-PD in non-CF bronchiectasis. All three techniques were well tolerated but ACBT-PD proved superior in terms of acute efficacy. Patient preference for treatment modality could not be predicted. PMID- 17416151 TI - Evaluating hospital at home and early discharge schemes for patients with an acute exacerbation of COPD. AB - Hospital at home and early discharge schemes for patients experiencing an acute exacerbation of their chronic obstructive pulmonary disease, appear to be an effective and safe option for selected patients and these services have become increasingly common. Here we discuss the evaluation of such schemes including: the rationale for evaluation; aspects of quality which might be considered for evaluation; the role of evaluation frameworks, quantitative and qualitative evaluation and steps in planning an evaluation. PMID- 17416152 TI - Respiratory health in Brazil. AB - According to the Brazilian Constitution each and every Brazilian has the right to free healthcare. Approximately 40% of the population have taken out a private or company funded health care program. Brazil is the world's largest producer of alcohol derived from sugar cane and the burning of the sugar cane leaves produces a large amount of fine and ultrafine particles what increases the relative risk of hospital admissions by children increases 3.5-fold due to respiratory diseases. Chronic obstructive pulmonary disease (COPD) ranks as the sixth greatest cause of death in Brazil, with a prevalence of 15.8% according to the fixed-ratio FEV1/FVC = 0.70 (PLATINO Study). The occurrence of wheezing in 13-14 year-old children over the last 12 months was 20%, while asthma diagnosed by a doctor was at around 10% (ISAAC Study). Mortality from asthma has decreased from 2.2/ 100,000 in 1980 to 1.37 in 1996, with currently 2500 deaths/year. Smoking is decreasing in Brazil; 32.6% of the adult population were smokers in 1989 while in 2002, a randomized survey carried out in over 107 Brazilian cities with a population of 200 thousand or over showed that the smoking population over 35 years of age had decreased to 24% (17% of overall population). Approximately one third of the Brazilian population has been infected by the tuberculosis bacillus and estimates agree that there are 116, 000 new cases per year but that only 90,000 are reported. Furthermore Brazil is ranked 14th in the world in the number of tuberculosis cases and the 22nd in incidence. A Family Health Program was created in 1994 and in 2005 there were 24,600 Family Health Teams functioning in Brazil, covering a total of 4986 cities, corresponding to approximately 45% of the 180 million Brazilian population. PMID- 17416153 TI - Thoracic oscillation, such as high frequency chest wall oscillators, provides oscillation directly to the chest wall. PMID- 17416154 TI - Is it necessary to use a noseclip in the performance of spirometry using a wedge bellows device? AB - There is no current consensus among published guidelines on whether noseclips are required during spirometry testing. This study investigated the effect of noseclips on spirometric measurements in patients with a range of disease. Fifty two patients (30 male; mean age 58.0 years, range 19-78; mean FEV1 82.6% predicted, range 23.8-128.3%) performed measurements of VC, FVC and FEV1 according to ARTP/BTS guidelines (1994) using a wedge bellows spirometer (Vitalograph Model S, Bucks, U.K.). All patients performed two sets of measurements (with and without noseclips) in random order (Group 1 = noseclips first; n=30; Group 2 (without noseclips first, n=22). Tests were conducted by qualified physiologists. Measurements obtained with and without the use of noseclips were similar (mean differences FEV1 -0.030 L SD 0.210 and -0.005 L SD 0.093 for Groups 1 and 2 respectively; FVC -0.007 L SD 0.109 and -0.040 L SD 0.117; VC 0.036L SD 0.137 and -0.040 L SD 0.150) and were not dependent on patient group or previous test experience. Four patients had differences outside the 95% confidence limits for each parameter. There were no significant correlations between the differences with and without noseclips and severity of lung disease, age, smoking history, BMI or lung volume (all P > 0.100). The within patient coefficient of variation did not depend on the testing method. Use of noseclips during spirometry does not systematically affect the results obtained or the within-subject repeatability. Marked individual differences highlight the importance of maintaining consistency in the method adopted for a particular patient. PMID- 17416155 TI - Bottom-up gaze shifts and fixations learning by imitation. AB - The ability to follow the gaze of conspecifics is a critical component in the development of social behaviors, and many efforts have been directed to studying the earliest age at which it begins to develop in infants. Developmental and neurophysiological studies suggest that imitative learning takes place once gaze following abilities are fully established and joint attention can support the shared behavior required by imitation. Accordingly, gaze-following acquisition should be precursory to most machine learning tasks, and imitation learning can be seen as the earliest modality for acquiring meaningful gaze shifts and for understanding the structural substrate of fixations. Indeed, if some early attentional process, based on a suitable combination of gaze shifts and fixations, could be learned by the robot, then several demonstration learning tasks would be dramatically simplified. In this paper, we describe a methodology for learning gaze shifts based on imitation of gaze following with a gaze machine, which we purposefully introduced to make the robot gaze imitation conspicuous. The machine allows the robot to share and imitate gaze shifts and fixations of a caregiver through a mutual vergence. This process is then suitably generalized by learning both the scene salient features toward which the gaze is directed and the way saccadic programming is attained. Salient features are modeled by a family of Gaussian mixtures. These together with learned transitions are generalized via hidden Markov models to account for humanlike gaze shifts allowing to discriminate salient locations. PMID- 17416156 TI - Skill acquisition through program-level imitation in a real-time domain. AB - This paper presents an imitation learning system capable of learning tasks in a complex dynamic real-time environment. In this paper, we argue that social learning should be thought of as a special case of general skill learning, and that the biases it presents to the skill learning problem radically simplify learning for species with sufficient innate predisposition to harness this power. We decompose skill learning into four subproblems, then show how a modification of Roy's CELL system can address all these problems simultaneously. Our system is demonstrated working in the domain of a real-time virtual-reality game, Unreal Tournament. PMID- 17416157 TI - On learning, representing, and generalizing a task in a humanoid robot. AB - We present a programming-by-demonstration framework for generically extracting the relevant features of a given task and for addressing the problem of generalizing the acquired knowledge to different contexts. We validate the architecture through a series of experiments, in which a human demonstrator teaches a humanoid robot simple manipulatory tasks. A probability-based estimation of the relevance is suggested by first projecting the motion data onto a generic latent space using principal component analysis. The resulting signals are encoded using a mixture of Gaussian/Bernoulli distributions (Gaussian mixture model/Bernoulli mixture model). This provides a measure of the spatio-temporal correlations across the different modalities collected from the robot, which can be used to determine a metric of the imitation performance. The trajectories are then generalized using Gaussian mixture regression. Finally, we analytically compute the trajectory which optimizes the imitation metric and use this to generalize the skill to different contexts. PMID- 17416158 TI - Correspondence mapping induced state and action metrics for robotic imitation. AB - This paper addresses the problem of body mapping in robotic imitation where the demonstrator and imitator may not share the same embodiment [degrees of freedom (DOFs), body morphology, constraints, affordances, and so on]. Body mappings are formalized using a unified (linear) approach via correspondence matrices, which allow one to capture partial, mirror symmetric, one-to-one, one-to-many, many-to one, and many-to-many associations between various DOFs across dissimilar embodiments. We show how metrics for matching state and action aspects of behavior can be mathematically determined by such correspondence mappings, which may serve to guide a robotic imitator. The approach is illustrated and validated in a number of simulated 3-D robotic examples, using agents described by simple kinematic models and different types of correspondence mappings. PMID- 17416159 TI - A developmental roadmap for learning by imitation in robots. AB - In this paper, we present a strategy whereby a robot acquires the capability to learn by imitation following a developmental pathway consisting on three levels: 1) sensory-motor coordination; 2) world interaction; and 3) imitation. With these stages, the system is able to learn tasks by imitating human demonstrators. We describe results of the different developmental stages, involving perceptual and motor skills, implemented in our humanoid robot, Baltazar. At each stage, the system's attention is drawn toward different entities: its own body and, later on, objects and people. Our main contributions are the general architecture and the implementation of all the necessary modules until imitation capabilities are eventually acquired by the robot. Also, several other contributions are made at each level: learning of sensory-motor maps for redundant robots, a novel method for learning how to grasp objects, and a framework for learning task description from observation for program-level imitation. Finally, vision is used extensively as the sole sensing modality (sometimes in a simplified setting) avoiding the need for special data-acquisition hardware. PMID- 17416160 TI - Incremental learning of tasks from user demonstrations, past experiences, and vocal comments. AB - Since many years the robotics community is envisioning robot assistants sharing the same environment with humans. It became obvious that they have to interact with humans and should adapt to individual user needs. Especially the high variety of tasks robot assistants will be facing requires a highly adaptive and user-friendly programming interface. One possible solution to this programming problem is the learning-by-demonstration paradigm, where the robot is supposed to observe the execution of a task, acquire task knowledge, and reproduce it. In this paper, a system to record, interpret, and reason over demonstrations of household tasks is presented. The focus is on the model-based representation of manipulation tasks, which serves as a basis for incremental reasoning over the acquired task knowledge. The aim of the reasoning is to condense and interconnect the data, resulting in more general task knowledge. A measure for the assessment of information content of task features is introduced. This measure for the relevance of certain features relies both on general background knowledge as well as task-specific knowledge gathered from the user demonstrations. Beside the autonomous information estimation of features, speech comments during the execution, pointing out the relevance of features are considered as well. The results of the incremental growth of the task knowledge when more task demonstrations become available and their fusion with relevance information gained from speech comments is demonstrated within the task of laying a table. PMID- 17416161 TI - Global optimization with multivariate adaptive regression splines. AB - This paper presents a novel procedure for approximating the global optimum in structural design by combining multivariate adaptive regression splines (MARS) with a response surface methodology (RSM). MARS is a flexible regression technique that uses a modified recursive partitioning strategy to simplify high dimensional problems into smaller yet highly accurate models. Combining MARS and RSM improves the conventional RSM by addressing highly nonlinear high-dimensional problems that can be simplified into lower dimensions, yet maintains a low computational cost and better interpretability when compared to neural networks and generalized additive models. MARS/RSM is also compared to simulated annealing and genetic algorithms in terms of computational efficiency and accuracy. The MARS/RSM procedure is applied to a set of low-dimensional test functions to demonstrate its convergence and limiting properties. PMID- 17416162 TI - Energy-based control of a haptic device using brakes. AB - This paper proposes an energy-based control method of a haptic device with electric brakes. Unsmooth motion is frequently observed in a haptic system using brakes during a wall-following task. Since it is generally known that a haptic system using brakes is passive due to brake's characteristics, its energy behavior has seldom been investigated. However, force distribution at the end effector reveals that the unsmooth motion of a haptic system using brakes represents active behavior of the system in the specific direction. A force control scheme is proposed that computes the gain for smooth motion by considering the energy behavior of a system. Experiments show that smooth wall following is possible with a proposed force control scheme. PMID- 17416163 TI - Stemming via distribution-based word segregation for classification and retrieval. AB - A novel corpus-based method for stemmer refinement, which can provide improvement in both classification and retrieval, is described. The method models the given words as generated from a multinomial distribution over the topics available in the corpus and includes a procedurelike sequential hypothesis testing that enables grouping together distributionally similar words. The system can refine any stemmer, and its strength can be controlled with parameters that reflect the amount of tolerance to be allowed in computing the similarity between the distributions of two words. Although obtaining the morphological roots of the given words is not the primary objective, the algorithm automatically does that to some extent. Despite a huge reduction in dictionary size, classification accuracies are seen to improve significantly when the proposed system is applied on some existing stemmers for classifying 20 Newsgroups and WebKB data. The refinements obtained are also suitable for cross-corpus stemming. Regarding retrieval, its superiority is extensively demonstrated with respect to four existing methods. PMID- 17416164 TI - Navigating a mobile robot by a traversability field histogram. AB - This paper presents an autonomous terrain navigation system for a mobile robot. The system employs a two-dimensional laser range finder (LRF) for terrain mapping. A so-called "traversability field histogram" (TFH) method is proposed to guide the robot. The TFH method first transforms a local terrain map surrounding the robot's momentary position into a traversability map by extracting the slope and roughness of a terrain patch through least-squares plane fitting. It then computes a so-called "polar traversability index" (PTI) that represents the overall difficulty of traveling along the corresponding direction. The PTIs are represented in a form of histogram. Based on this histogram, the velocity and steering commands of the robot are determined. The concept of a virtual valley and an exit condition are proposed and used to direct the robot such that it can reach the target with a finite-length path. The algorithm is verified by simulation and experimental results. PMID- 17416165 TI - An automatically tuning intrusion detection system. AB - An intrusion detection system (IDS) is a security layer used to detect ongoing intrusive activities in information systems. Traditionally, intrusion detection relies on extensive knowledge of security experts, in particular, on their familiarity with the computer system to be protected. To reduce this dependence, various data-mining and machine learning techniques have been deployed for intrusion detection. An IDS is usually working in a dynamically changing environment, which forces continuous tuning of the intrusion detection model, in order to maintain sufficient performance. The manual tuning process required by current systems depends on the system operators in working out the tuning solution and in integrating it into the detection model. In this paper, an automatically tuning IDS (ATIDS) is presented. The proposed system will automatically tune the detection model on-the-fly according to the feedback provided by the system operator when false predictions are encountered. The system is evaluated using the KDDCup'99 intrusion detection dataset. Experimental results show that the system achieves up to 35% improvement in terms of misclassification cost when compared with a system lacking the tuning feature. If only 10% false predictions are used to tune the model, the system still achieves about 30% improvement. Moreover, when tuning is not delayed too long, the system can achieve about 20% improvement, with only 1.3% of the false predictions used to tune the model. The results of the experiments show that a practical system can be built based on ATIDS: system operators can focus on verification of predictions with low confidence, as only those predictions determined to be false will be used to tune the detection model. PMID- 17416166 TI - Feasibility study on a hyperacuity device with motion uncertainty: two-point stimuli. AB - Can a device that can perform hyperacuity vision tasks be built? In this paper, a feasibility study based on separation discrimination is conducted. Two types of ideal detectors are considered. The first is the stimulus defined statistically (SDS) detector by Geisler and Davilla. The second is one that estimates the uncertainty and then takes out its effect. In the first method, an array of many ideal stimulus defined exactly (SDE) detectors covers uncertainties and forms the ideal SDS detector. When the separation distance between the SDE detectors is around 1 arcmin, the SDS detector can achieve nearly optimal performance. To cover the motion uncertainty with nearly optimal performance, the SDE detector at each position needs to cover 16 directions, and at each direction, it needs to cover speeds with an increment of 0.5 degrees/s. Typically, the SDS detector needs 7776 SDE detectors to deal with a speed up to 2 degrees/s stimulus movement with a randomly selected direction and a 9 min x 9 min position uncertainty region. This ideal observer can achieve a hyperacuity threshold of 2-4 arcsec. Its threshold is almost constant over the range of speeds covered by the SDS detector. Using the second method, position estimation and motion tracking capability is examined. With perfect position estimation and motion tracking, the SDS detector can be reduced to a single SDE detector that is tuned to correct position and motion direction and speed. Two lower bounds on the estimation variance are examined, namely: 1) the Cramer-Rao bound and 2) the Ziv-Zakai bound. The results showed that if an estimation algorithm that can achieve the performance of bounds can be found, then the second method could achieve a hyperacuity capability of 1 s or less. The human visual system may more likely adopt the first method, but the second is simpler to use in building up a device using microprocessors. In this paper, dot-pair templates are used, which precisely match to dot-pair stimuli, to compute the likelihood and make a decision. The main difference between this paper and that of Geisler and Davilla lies in using a discrete sum over an array of SDE detectors to closely approximate continuous integration over the uncertain region, which makes it much easier in hardware implementation. PMID- 17416167 TI - A study on expertise of agents and its effects on cooperative Q-learning. AB - Cooperation in learning (CL) can be realized in a multiagent system, if agents are capable of learning from both their own experiments and other agents' knowledge and expertise. Extra resources are exploited into higher efficiency and faster learning in CL as compared to that of individual learning (IL). In the real world, however, implementation of CL is not a straightforward task, in part due to possible differences in area of expertise (AOE). In this paper, reinforcement-learning homogenous agents are considered in an environment with multiple goals or tasks. As a result, they become expert in different domains with different amounts of expertness. Each agent uses a one-step Q-learning algorithm and is capable of exchanging its Q-table with those of its teammates. Two crucial questions are addressed in this paper: "How the AOE of an agent can be extracted?" and "How agents can improve their performance in CL by knowing their AOEs?" An algorithm is developed to extract the AOE based on state transitions as a gold standard from a behavioral point of view. Moreover, it is discussed that the AOE can be implicitly obtained through agents' expertness in the state level. Three new methods for CL through the combination of Q-tables are developed and examined for overall performance after CL. The performances of developed methods are compared with that of IL, strategy sharing (SS), and weighted SS (WSS). Obtained results show the superior performance of AOE-based methods as compared to that of existing CL methods, which do not use the notion of AOE. These results are very encouraging in support of the idea that "cooperation based on the AOE" performs better than the general CL methods. PMID- 17416168 TI - State-based control of fuzzy discrete-event systems. AB - To effectively represent possibility arising from states and dynamics of a system, fuzzy discrete-event systems (DESs) as a generalization of conventional DESs have been introduced recently. Supervisory-control theory based on event feedback has been well established for such systems. Noting that the system state description, from the viewpoint of specification, seems more convenient, we investigate the state-based control of fuzzy DESs in this paper. An approach to finding all fuzzy states that are reachable by controlling the system is presented first. After introducing the notion of controllability for fuzzy states, a necessary and sufficient condition for a set of fuzzy states to be controllable is then provided. It was also found that event- and state-based controls are not equivalent, and the relationship between them was further discussed. Finally, we examine the possibility of driving a fuzzy DES under control from a given initial state to a prescribed set of fuzzy states and then keeping it there indefinitely. PMID- 17416169 TI - Reinforcement learning neural-network-based controller for nonlinear discrete time systems with input constraints. AB - A novel adaptive-critic-based neural network (NN) controller in discrete time is designed to deliver a desired tracking performance for a class of nonlinear systems in the presence of actuator constraints. The constraints of the actuator are treated in the controller design as the saturation nonlinearity. The adaptive critic NN controller architecture based on state feedback includes two NNs: the critic NN is used to approximate the "strategic" utility function, whereas the action NN is employed to minimize both the strategic utility function and the unknown nonlinear dynamic estimation errors. The critic and action NN weight updates are derived by minimizing certain quadratic performance indexes. Using the Lyapunov approach and with novel weight updates, the uniformly ultimate boundedness of the closed-loop tracking error and weight estimates is shown in the presence of NN approximation errors and bounded unknown disturbances. The proposed NN controller works in the presence of multiple nonlinearities, unlike other schemes that normally approximate one nonlinearity. Moreover, the adaptive critic NN controller does not require an explicit offline training phase, and the NN weights can be initialized at zero or random. Simulation results justify the theoretical analysis. PMID- 17416170 TI - An ensemble-based incremental learning approach to data fusion. AB - This paper introduces Learn++, an ensemble of classifiers based algorithm originally developed for incremental learning, and now adapted for information/data fusion applications. Recognizing the conceptual similarity between incremental learning and data fusion, Learn++ follows an alternative approach to data fusion, i.e., sequentially generating an ensemble of classifiers that specifically seek the most discriminating information from each data set. It was observed that Learn++ based data fusion consistently outperforms a similarly configured ensemble classifier trained on any of the individual data sources across several applications. Furthermore, even if the classifiers trained on individual data sources are fine tuned for the given problem, Learn++ can still achieve a statistically significant improvement by combining them, if the additional data sets carry complementary information. The algorithm can also identify-albeit indirectly-those data sets that do not carry such additional information. Finally, it was shown that the algorithm can consecutively learn both the supplementary novel information coming from additional data of the same source, and the complementary information coming from new data sources without requiring access to any of the previously seen data. PMID- 17416171 TI - Developmental word grounding through a growing neural network with a humanoid robot. AB - This paper presents an unsupervised approach of integrating speech and visual information without using any prepared data. The approach enables a humanoid robot, Incremental Knowledge Robot 1 (IKR1), to learn word meanings. The approach is different from most existing approaches in that the robot learns online from audio-visual input, rather than from stationary data provided in advance. In addition, the robot is capable of learning incrementally, which is considered to be indispensable to lifelong learning. A noise-robust self-organized growing neural network is developed to represent the topological structure of unsupervised online data. We are also developing an active-learning mechanism, called "desire for knowledge," to let the robot select the object for which it possesses the least information for subsequent learning. Experimental results show that the approach raises the efficiency of the learning process. Based on audio and visual data, they construct a mental model for the robot, which forms a basis for constructing IKRI's inner world and builds a bridge connecting the learned concepts with current and past scenes. PMID- 17416172 TI - Segmentation-assisted detection of dirt impairments in archived film sequences. AB - In this correspondence, a novel segmentation-assisted method for film-dirt detection is proposed. We exploit the fact that film dirt manifests in the spatial domain as a cluster of connected pixels whose intensity differs substantially from that of its neighborhood, and we employ a segmentation-based approach to identify this type of structure. A key feature of our approach is the computation of a measure of confidence attached to detected dirt regions, which can be utilized for performance fine tuning. Another important feature of our algorithm is the avoidance of the computational complexity associated with motion estimation. Our experimental framework benefits from the availability of manually derived as well as objective ground-truth data obtained using infrared scanning. Our results demonstrate that the proposed method compares favorably with standard spatial, temporal, and multistage median-filtering approaches and provides efficient and robust detection for a wide variety of test materials. PMID- 17416173 TI - Emergent behaviors of protector, refugee, and aggressor swarms. AB - Simple rules, when executed by individual agents in a large group, or swarm, can lead to complex behaviors that are often difficult or impossible to predict knowing only the rules. However, aggregate behavior is not always unpredictable even for swarm models said to be beyond analysis. For the class of swarming algorithms examined herein, we analytically identify several possible emergent behaviors and their underlying causes: clustering, drifting, and explosion. They also analyze the likelihood of these behaviors emerging from randomly selected swarm configurations and present a few examples. The analytic results are illustrated via simulation. PMID- 17416174 TI - A potential field model using generalized sigmoid functions. AB - The lack of a potential field model capable of providing accurate representations of objects of arbitrary shapes is considered one major limitation in applying the artificial potential field method in many practical applications. In this correspondence, we propose a potential function based on generalized sigmoid functions. The generalized sigmoid model can be constructed from combinations of implicit primitives or from sampled surface data. The constructed potential field model can achieve an accurate analytic description of objects in two or three dimensions and requires very modest computation at run time. In this correspondence, applications of the generalized sigmoid model in path-planning tasks for mobile robots and in haptic feedback tasks are presented. The validation results in this correspondence show that the model can effectively allow the user or mobile robot to avoid penetrations of obstacles while successfully accomplishing the task. PMID- 17416175 TI - An intelligent subtitle detection model for locating television commercials. AB - A strategy for locating television (TV) commercials in TV programs is proposed. Based on the observation that most TV commercials do not have subtitles, the first stage exploits six subtitle constraints and an adaptive neurofuzzy inference system model to determine whether a frame contains a subtitle or not. The second stage involves locating the mark-in/mark-out points using a genetic algorithm. An interactive user interface allows users to efficiently identify and fine-tune the exact boundaries separating the commercials from the program content. Furthermore, erroneous boundaries are manually corrected. Experimental results show that the precision rate and recall rates exceed 90%. PMID- 17416176 TI - Nonfragile Hinfinity control for uncertain neutral systems with time-varying delays via the LMI optimization approach. AB - The nonfragile Hinfinity control problem for a neutral system with time-varying delays is considered. Delay-dependent criteria are proposed to guarantee the stabilization and disturbance attenuation of systems. A linear matrix inequality optimization approach is used to solve the nonfragile Hinfinity control problem. Nonfragile Hinfinity control for an unperturbed neutral system is investigated first. Then, nonfragile Hinfinity control for an uncertain neutral system is derived directly from the unperturbed case. Finally, two examples are illustrated to show the improvement of this correspondence. PMID- 17416177 TI - [Sign of the times]. PMID- 17416178 TI - [Management of problem wounds--a case report: amputation could be avoided]. PMID- 17416179 TI - [An efficient method for managing problem wounds: maggot therapy in biosurgery]. PMID- 17416180 TI - [Therapy option after kidney transplantation--2: Improving activities at the beginning of a new life]. PMID- 17416181 TI - [Problems from general practice--solutions for general practice: preoperative fasting times]. PMID- 17416182 TI - [Pressure ulcer prevention in German hospitals. An analysis of nursing practice with regard to guideline recommendations]. AB - Pressure ulcers are still a common health problem. Prevention and therapy of pressure ulcers require extensive nursing resources. Based on epidemiological data the use of preventive interventions and devices in hospital patients at risk was analysed. One object of this study was to compare the nursing practice with the recommendations given by a national practice guideline about pressure ulcer prevention by the German Network for Quality Development in Nursing (DNQP). Since 2002 each year a study has been conducted in order to investigate the prevalence of pressure ulcers in German hospitals. For the present study data of 9159 patients at pressure ulcer risk collected in three years (from 2002 to 2004) were analysed. RESULTS: In the course of the investigated period interventions and devices were increasingly adjusted to the recommendations of the guideline. This trend appears to be slightly, but it is noticeable. The number of process requirements realised for each patient increased particularly if patients showed a high risk or a pressure ulcer was already present. To improve the quality of care preferably all process criteria according to the guideline should be considered if preventive measures are planned. PMID- 17416183 TI - [Comment about the policy statement of the MDS. A position statement of the assessment group of the University Witten/Herdecke with regard to nursing documentation]. AB - In 2005 the Medical Advisory Service of Social Health Insurance (MDS) in Germany published a policy statement with regard to the nursing process and documentation. According to the intention of the association, this statement should be considered as recommendations which are able to improve nursing practice and to contribute to streamlining of bureaucracy in nursing care. Recognising the broad impact of this publication on nursing institutions, a working group on nursing assessment of the University Witten/Herdecke conducted a critical review of the statement. Significant criteria for evaluation were the primary role of nursing documentation, quality requirements for the documentation as well as recent scientific results concerning the implementation of nursing process and assessment-based nursing diagnoses. The review revealed that the statement lacks of a clear rationale and its content appears to be merely research-based. Therefore it has to be questioned if the publication will accomplish the claimed effects. In fact, future quality criteria for health care are to be developed independently on the basis of scientific results and in consideration of the experiences of all concerned social groups. PMID- 17416184 TI - [Bremen International Study Curriculum for Nursing and Health Management: 2nd graduate survey: diploma--and then what?]. PMID- 17416185 TI - [Hospital certification according to proCum inkl. KTOQ (pCC certification): responsible for special charge]. PMID- 17416186 TI - [Decubitus ulcer prevention expert standard--excerpts from implementation: on the path to continuous improvements]. PMID- 17416187 TI - [Clinical case management in an Australian clinic: planning discharge already at admission]. PMID- 17416188 TI - [Mandatory treatment of mentally competent patients: patient's will has priority]. PMID- 17416189 TI - [From case to case: deciding moments]. PMID- 17416190 TI - Pathophysiological actions of protease activated receptors (PARs). AB - Serine proteases such as thrombin, mast cell tryptase, trypsin, or cathepsin G, for example, are highly active mediators with diverse biological activities. So far, proteases have been considered to act primarily as degradative enzymes in the extracellular space. However, their biological actions in tissues and cells suggest important roles as a part of the body's hormonal communication system during inflammation and immune response. These effects can be attributed to the activation of a new subfamily of G protein-coupled receptors, termed protease activated receptors (PARs). Four members of the PAR family have been cloned so far. Thus, certain proteases act as signaling molecules that specifically regulate cells by activating PARs. After stimulation, PARs couple to various G proteins and activate signal transduction pathways resulting in the rapid transcription of genes that are involved in inflammation. For example, PARs are widely expressed by cells involved in immune responses and inflammation, regulate endothelial-leukocyte interactions, and modulate the secretion of inflammatory mediators or neuropeptides. Together, the PAR family necessitates a paradigm shift in thinking about hormone action, to include proteases as key modulators of biological function. Novel compounds that can modulate PAR function may be potent candidates for the treatment of inflammatory or immune diseases. PMID- 17416191 TI - An enantiospecific HPLC method for the determination of (S)-enantiomer impurities in (R)-tolterodine tartarate. AB - A high-performance liquid chromatographic method was developed for the separation of the enantiomers of tolterodine tartarate. The proposed method was applied to the determination of (S)-isomer in (R)-tolterodine tartarate, and satisfactory results were obtained. The enantiomers of tolterodine tartarate were separated on a Chiralpak AD-H (250 mm x 4.6 mm) column containing amylose tris-(3,5 dimethylphenyl-carbamate) at room temperature. The mobile phase consisted of n hexane and isopropyl alcohol in the ratio of 85:15 (v/v) with 0.075% triethylamine (TEA) and 0.05% trifluoroacetic acid (TFA) as the additive. The flow rate was kept at 0.5 ml/min, and UV detection wavelength was set at 283 nm. The calibration curves of (S)-enantiomer in the concentration range from 0.05 microg/ml to 1 microg/ml range were linear. The relative standard deviations of within-day and between-day were less than 2% (n = 3). The limit of detection (LOD) was 0.75 ng (S/N = 3) and the limit of quantification (LOQ) was 0.05 microg/ml (RSD < 4.1%, n = 3). The determination recoveries of the (S)-enantiomer were in the range of 98.2-104.8%. The results demonstrated that the developed HPLC method was a reliable, simple technique and was applicable to the purity determination of (R)- tolterodine tartarate. PMID- 17416192 TI - A simple HPLC-UV method for the determination of dimenhydrinate and related substances--dentification of an unknown impurity. AB - During the revision of the dimenhydrinate monograph of the European Pharmacopoeia a HPLC-UV method was developed. The procedure described allows a qualitative and quantitative determination of both dimenhydrinate compounds and of thirteen related substances. Furthermore a hitherto unknown impurity was identified and integrated into the purity check. Also 18 samples of dimenhydrinate have been tested. Thereby the relevant impurities of dimenhydrinate could be nominated and quantified. PMID- 17416193 TI - Simultaneous determination of trimethoprim and sulphamethoxazole in veterinary formulations by chromatographic multivariate methods. AB - A comparative chromatographic study was developed for the simultaneous quantitative resolution of trimethoprim (TMP) and sulphamethoxazole (SMX) in veterinary formulations. Multi-wavelength chromatograms were recorded by using diode array detector (DAD) system at the five-wavelength set consisting of 220, 230, 240, 250 and 260 nm. In the first step, five different calibration equations at the above wavelengths for each drug were obtained by using the relationship between concentration and peak area. These calibration graphs were used for the quantitative evaluation of TMP and SMX in samples. These single-wavelength applications were called traditional LC method. In the second step, principal component regression (PCR) and partial least-squares (PLS) calibrations were applied to the above mentioned multi-wavelength chromatograms. The amount of two investigated drugs in samples was determined by the constructed PCR and PLS calibrations. The experimental results obtained from each single-wavelength calibration graph were compared with those obtained by the chemometric approaches and chromatographic multivariate approaches give successful results more than traditional LC method. PMID- 17416194 TI - Penetrometry and estimation of the flow rate of powder excipients. AB - In this work, penetrometry with a sphere was employed to study the flow properties of non-consolidated pharmaceutical powder excipients: sodium chloride, sodium citrate, boric acid, and sorbitol. In order to estimate flow rate, the pressure of penetration in Pascals was used. Penetrometry measurement with a sphere requires modification of the measurement container, in particular by decreasing the diameter of the container, to prevent undesirable movement of material in a direction opposite to that in which the sphere penetrates. Thus penetrometry by a sphere seems to be similar to indentation by the Brinell hardness tester. The pressure of penetration was determined from the depth of penetration by analogy with the Brinell hardness number and an equation for the inter conversion of the two variables is presented. The penetration pressure allowed direct estimation of the flow rate only for those powder excipients with a size fraction in the range of 0.250-0.630 mm. Using the ratio of penetration pressure to bulk density, a polynomial quadratic equation was generated from which the flow rates for the group of all tested powders could be estimated. Finally, if the inverse ratio of bulk density and penetration pressure was used as an independent variable, the flow rate could be estimated by linear regression with the coefficient of determination r2 = 0.9941. In conclusion, using sphere penetrometry, the flow properties of non-consolidated powder samples could be investigated by indentation. As a result, a linear regression in which the flow rate was directly proportional to the powder bulk density and inversely proportional to the penetration pressure could be best recommended for the estimation of the flow rate of powder excipients. PMID- 17416195 TI - Prediction of the optimized solvent composition for solubilization of drugs in water-cosolvent mixtures. AB - The capability of the Jouyban-Acree model for predicting the optimized solvent composition of binary solvents for solubilization of drugs is shown employing solubility of drugs in aqueous mixtures of dioxane, ethanol and polyethylene glycol 400. The established model constants of the Jouyban-Acree model and solubility of drugs in water and cosolvent are used to predict the maximum solubility of in the binary solvent mixture (log Xm(max)) and the corresponding solvent composition (f1,max). The accuracy of the predicted log Xm(max) and f1,max is studied using average absolute error (AAE) of predicted and observed values. The AAEs were 0.10 +/- 0.12 and 0.08 +/- 0.10, respectively for log Xm(max) and f1,max. The method provided acceptable predictions and is recommended for practical applications. The main advantage of the proposed method is its extension to temperatures higher/lower than room temperature. PMID- 17416196 TI - Evaluation of preservative systems in opthalmic suspension of polymyxin B and dexamethasone by linear regression. AB - An ophthalmic suspension of polymyxin B and dexametasone with 13 different preservative systems was evaluated by a linear regression method for rapid screening, using D-values. The products were challenged against Pseudomonas aeruginosa, Burkholderia cepacia, Staphylococcus aureus, Candida albicans and Aspergillus niger. Since linear regression showed correlation coefficients higher than 0.84 the decimal reduction time (D-value) was then calculated. The comparison of D-values obtained for each microorganism and each product led to the selection of the formula 9 = 10, which was selected to be submitted to the efficacy of antimicrobial preservation according to the European Pharmacopoeia. PMID- 17416197 TI - Cytotoxicity evaluation of anionic nanoliposomes and nanolipoplexes prepared by the heating method without employing volatile solvents and detergents. AB - Submicron lipid vesicles (nanoliposomes) are being used as carriers of bioactive compounds. In addition, complexes of nanoliposomes and nucleic acids (nanolipoplexes) are promising tools for the treatment of cancer, and viral and genetic disorders. Toxicity of some of these formulations, however, still remains a concern in their clinical utilisation. To address this problem, anionic liposomes were prepared by two different techniques, the conventional thin-film method, and the heating method (HM), in which no volatile organic solvent or detergent is used. An anionic nanolipoplex was constructed by incorporating plasmid DNA (pcDNA3.1/His B/lacZ) into the HM-nanoliposomes by the mediation of calcium. The toxicity of the nanoliposomes, with and without plasmid and Ca2+, was assessed using a human bronchial epithelial cell line (16HBE14o-) in the presence of serum. Cytotoxicity evaluations performed by two different assays (i.e. NRU and MTT) indicated that HM-nanoliposomes were completely non-toxic in the cell-line tested, whereas conventional liposomes revealed significant levels of toxicity. This may be due to the presence of trace amounts of chloroform and/or methanol applied during their preparation. Similar results were obtained for different sizes of lipid vesicles (prepared by 100 nm and 400 nm pore-size filters). In addition, it was observed that incorporation of DNA (15 microg/ 285 microg lipid) and Ca2+ (50 mM) to the nanoliposomes did not have any effect on their cytotoxicities. These findings indicate that the HM-liposomes have great potential as non-toxic delivery vehicles in human gene therapy and drug delivery applications while liposomes made using organic solvents should be used with caution. PMID- 17416198 TI - Interaction of neomycin, tobramycin and amikacin with melanin in vitro in relation to aminoglycosides-induced ototoxicity. AB - The aim of this study was to examine in vitro the interaction between aminoglycoside antibiotics displaying adverse ototoxic effects and melanin which is a constituent of the inner ear. The binding of neomycin, tobramycin and amikacin to model synthetic melanin was studied. It has been demonstrated that all the investigated aminoglycosides form stable complexes with melanin biopolymer. The obtained results show that the amount of drug bound to melanin increases with the increase of initial drug concentration and the incubation time. An analysis of drugs binding to melanin by the use of Scatchard plots has shown that at least two classes of independent binding sites must be implicated in the studied aminoglycoside antibiotic-melanin complexes formation: strong binding sites (n1) with the association constant K1 approximately 0.2-2.0 x 10(5) M(-1) and weak binding sites (n2) with K2 approximately 1.0-4.9 x 10(3) M(-1). Based on the values of association constants the following order of drugs affinity to DOPA-melanin was found: tobramycin > amikacin >> neomycin. The ability of the analyzed aminoglycoside antibiotics to form complexes with melanin in vitro may be one of the reasons for their ototoxicity in vivo, as a result of their accumulation in melanin in the inner ear. PMID- 17416199 TI - Apigenin increases osteoblastic differentiation and inhibits tumor necrosis factor-alpha-induced production of interleukin-6 and nitric oxide in osteoblastic MC3T3-E1 cells. AB - Many plant-derived substances have estrogenic activities. Due to their ability to bind the estrogen receptor, these compounds have the potential to counteract the deleterious effects of estrogen deficiency on bone. In this study, the effects of apigenin on the function of osteoblastic MC3T3-E1 cells and the production of local factors in osteoblasts were investigated. Apigenin (0.01 microM) significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells (P < 0.05). The effect of apigenin in increasing ALP activity and collagen content was completely prevented by the presence of 10(-6) M cycloheximide and 10 6 M tamoxifen, suggesting that apigenin's effect results from a newly synthesized protein component and might be partly involved in estrogen action. Locally derived mediators in bone play a crucial role in the regulation of bone remodeling, i.e., bone formation and bone resorption processes. The effect of apigenin on the TNF-alpha-induced production of IL-6 and nitric oxide (NO) in osteoblasts was examined. Treatment with apigenin (10 microM) decreased the TNF alpha-induced production of IL-6 and NO in osteoblasts. Taken together, these results suggest that apigenin may represent new pharmacological tools for the treatment of osteoporosis. PMID- 17416200 TI - Effect of aloe vera leaf gel extract on membrane bound phosphatases and lysosomal hydrolases in rats with streptozotocin diabetes. AB - Diabetes mellitus is known to promote deterioration of membrane function and impair intra cellular metabolism in the organism. The aim of the present study was to examine the effect of the ethanolic extract from Aloe vera leaf gel on membrane bound phosphatases and lysosomal hydrolases in the liver and kidney of streptozotocin (STZ)-induced diabetic rats. The rats treated with STZ showed significant alterations in the activities of membrane bound phosphatases and lysosomal hydrolases in the liver and kidney. Oral administration of Aloe vera gel extract at a dose of 300 mg/kg body weight/day to STZ-induced diabetic rats for a period of 21 days significantly restored the alterations in enzymes activity to near normalcy. These results were compared with glibenclamide, a reference drug. Thus, the present study confirms that Aloe vera gel extract possesses a significant beneficial effect on membrane bound phosphatases and lysosomal hydrolases. PMID- 17416201 TI - Metabolism of parthenin by Beauveria bassiana ATCC 7159. AB - Two new metabolites were isolated from the biotransformation reaction of parthenin by Beauveria bassiana ATCC 7159 along with the known naturally occurring sesquiterpene lactones hymenolin and dihydrocoronopilin. The new metabolites: are 3beta,4beta,6beta-trihydroxy-10alphaH,11alpha-methyl-ambrosa-1 en-12-oic acid-gamma-lactone and 3alpha,6beta-dihydroxy-4beta-hydroperoxy 10alphaH, 11alpha-methyl-ambrosa-1-en-12-oic acid-gamma-lactone. The structure elucidation of these compounds was achieved by different spectroscopic methods. PMID- 17416202 TI - Flavan-3-ols and procyanidins from the bark of Salix purpurea L. AB - From a commercial aqueous ethanolic extract obtained from the bark of Salix purpurea L. the flavan-3-ols catechin, epicatechin, gallocatechin, catechin-3-O (1-hydroxy-6-oxo-2-cyclohexene-1-carboxylic acid)-ester, the dimeric procyanidins B1, B3 and the trimeric procyanidins epicatechin-(4B-->8)-catechin-(4alpha-->8) catechin and epicatechin-(4beta-->8)-epicatechin-(4beta-->8)-catechin were isolated. Structure elucidation was performed by NMR, CD, MS, degradation and optical rotation methods. A fraction containing higher oligomeric procyanidins was investigated by 13C NMR. Data indicate an average degree of oligomerization of 4 to 5 flavan-3-ol units with dihydroxylated B-rings and predominance of 2,3 cis-stereochemistry. PMID- 17416203 TI - Influence of ethanol water-content on gatifloxacin recrystallisations. AB - Gatifloxacin is a flouroquinolone anti-bacterial agent. This study proved that even the presence of small amounts of water in the recrystallisation medium influenced the product obtained. Different crystal forms were produced from various binary mixtures of ethanol and water. Where mixtures had a water content greater than (or equal to) 50% v/v, only hydrated crystal forms were produced and no ethanol was incorporated into the lattice. PMID- 17416204 TI - Prolonged cytotoxic effect of aqueous extracts from dried viscum album on bladder cancer cells. AB - Aqueous extracts from whole dried mistletoe (Viscum album L., Iscucin) are often used in anti-cancer treatment. We studied the effect of extracts obtained from mistletoe bushes that grew on different host trees on bladder cancer cells by means of MTT-colorimetric cell proliferation/survival assays. The extracts possessed concentration-dependent cytotoxic properties whose extent varied with the host tree, but did not always correlate with the corresponding mistletoe lectin content. A 2-hours treatment of bladder cancer cells triggered a later, strong cytotoxic effect. This prolonged effect suggests that instillation with Iscucin has therapeutic potential for bladder cancer patients. PMID- 17416205 TI - Prednisolone concentration in the cochlea of patients with perilymph fistula. AB - After iv administration of 200 mg prednisolone in patients with perilymph fistula, concentrations of the drug in the cochlea were determined. A specially adapted LC method was used for analysis. Mean concentrations of prednisolone in the perilymphe reached 95 ng/ml after 15-25 min, and 338 ng/ml after 30-45 min. The values reached 8 and 41% of the corresponding serum concentrations, respectively. PMID- 17416206 TI - Sphingosine-1-phosphate lyase in immunity and cancer: silencing the siren. AB - Sphingosine-1-phosphate (S1P) is a bioactive lipid that promotes cell survival, proliferation and migration, platelet aggregation, mediates ischemic preconditioning, and is essential for angiogenesis and lymphocyte trafficking. Sphingosine-1-phosphate lyase (SPL) is the enzyme responsible for the irreversible degradation of S1P and is, thus, in a strategic position to regulate these same processes by removing available S1P signaling pools, that is, silencing the siren. In fact, recent studies have implicated SPL in the regulation of immunity, cancer surveillance and other physiological processes. Here, we summarize the current understanding of SPL function and regulation, and discuss how SPL might facilitate cancer chemoprevention and serve as a target for modulation of immune responses in transplantation settings and in the treatment of autoimmune disease. PMID- 17416207 TI - Predictive value of local and core laboratory echocardiographic assessment of cardiac function in patients with chronic stable angina: The ACTION study. AB - AIMS: To evaluate the relationship between echocardiographic cardiac function and outcome in patients with stable symptomatic angina. METHODS: Baseline echo left ventricular ejection fraction and volume data measured in a central laboratory was available for 7016 patients (92% of the total) participating in the ACTION trial (A Coronary disease Trial Investigating Outcome with Nifedipine GITS). Ejection fraction was also measured by investigators. Evaluation of the different echocardiographic variables was based on adjusted hazard ratios comparing the unfavourable limit of the 90% range of the variable concerned to the favourable limit. RESULTS: The centrally measured ejection fraction was the most powerful predictor of all-cause death (adjusted hazard ratio=2.5), myocardial infarction, any stroke or transient ischaemic attack and overt heart failure (adjusted hazard ratio=4.5). The addition of either end systolic volume or end diastolic volume to ejection fraction did not materially affect the power of prediction. Compared to the central ejection fraction measurement, the investigator-measured ejection fraction was a less powerful predictor for all outcomes considered. CONCLUSION: Routine echocardiography carefully analysed by standardised methods provides useful prognostic information in patients with stable angina, including for total mortality. PMID- 17416208 TI - The role of comorbid psychiatric conditions in health status in epilepsy. AB - Comorbid psychiatric conditions are highly prevalent in patients with epilepsy, yet the long-term implications across multiple mental health conditions are poorly understood. We examined the association between psychiatric diagnoses and self-reported health status in veterans with epilepsy. ANCOVA models were used to derive adjusted SF-36V scores for individuals with epilepsy alone (N=7379) or with additional psychiatric conditions (N=6320): depression, schizophrenia, bipolar disorder, anxiety disorder, substance abuse, and posttraumatic stress disorder (PTSD). Compared with patients with epilepsy alone, scores of veterans with comorbid psychiatric diagnoses averaged 21% lower across all domains. Role Limitation scales exhibited the greatest decrement across domains. A PTSD diagnosis consistently corresponded to lower scores, followed by depression. Schizophrenia contributed the least detriment to perceived health status. Comorbid psychiatric conditions impart significant emotional and physical burdens, requiring timely recognition and treatment of these disorders. Patients with epilepsy are uniquely at risk for high physical-psychiatric comorbidity profiles, with concomitant losses in perceived health status. PMID- 17416209 TI - Health-related quality of life in adolescents with well-controlled epilepsy. AB - OBJECTIVE: The purpose of the work described in this article was to evaluate the health-related quality of life (HRQOL) of adolescents with well-controlled epilepsy. METHODS: The Quality of Life in Epilepsy Inventory for Adolescents 48 (QOLIE-AD-48) was completed by 71 subjects with uncomplicated epilepsy who had been seizure-free for more than a year. HRQOL was measured in eight domains: Health Perception, Epilepsy Impact, Memory/Concentration, Physical Functioning, Stigma, Social Support, School Behavior, and Attitudes toward Epilepsy. In addition, antiepileptic drug (AED) therapy and concern over seizures recurring were analyzed in relation to HRQOL. RESULTS: The mean HRQOL total scores were 83.9 for boys and 83.06 for girls. The highest scores were observed in the Physical Functioning and School Behavior domains; the lowest in the Attitudes toward Epilepsy domain. Girls reported more positive perceptions in the Stigma and Social Support domains. Greater intake of AEDs and concern over seizures recurring accounted for poorer HRQOL. CONCLUSIONS: Adolescents with favorable seizure control evaluated their HRQOL as satisfactory. However, they perceived their best adjustment to epilepsy to be in the Physical Functioning and School Behavior domains. AED intake and concern over seizures recurring were predictors of HRQOL in this group. PMID- 17416211 TI - MCT-1 oncogene downregulates p53 and destabilizes genome structure in the response to DNA double-strand damage. AB - Tumor suppressor p53 protein mediates checkpoint controls and the apoptotic program that are critical for maintaining genomic integrity and preventing tumorigenesis. Forced-induction of MCT-1 decreased p53 expression before and after genomic insults. While inhibiting protein synthesis, the levels of ubiquinated-p53 and the phospho-MDMA2 were significantly increased in ectopic MCT 1 cells. Abrogation of the proteosome degradation process attenuated p53 destabilization and p21 down-regulation by MCT-1. Concomitantly, MCT-1 overexpression enhanced the phosphorylation status of MAPK (ERK1/ERK2). While MCT 1 gene knockdown or MEK/ERK pathway inhibition dramatically reduced MAPK phosphorylation, the genotoxin-induced p53 and p21 production were noticeably elevated. Upon Etoposide treatment, ectopic MCT-1 cells relaxed S-phase and G2/M checkpoints followed by G1 phase progressing. Moreover, cells inducing with MCT-1 abridged accumulations of G2/M populations in the response to gamma-irradiation. The polyploidy (DNA content>4N) populations were increased in association with p53 loss in MCT-1 oncogenic cells. Alkaline comet assay validated that ectopic MCT-1 cells were less susceptibility to the genotoxicity. Furthermore, the allocation of nuclear MCT-1 induced by the genotoxic stress was moderately coincided with gamma-H2AX appearances. Throughout damage-repairing process, ectopic MCT-1 cells displayed many larger chromosomes and multiple chromosomal fusions compared to the controls that showed increase in chromosomal breaks/gaps and minute chromosomal fragments. Spectral karyotyping analysis precisely identified the acquisition of a single extra copy of chromosome 14 together with a complex genome organizations in ectopic MCT-1 cells, including extra copies of chromosome segments that had been translocated to derivative chromosomes 6 [der(6)] and 9 [der(9)]. In conclusion, MCT-1 deregulates p53-p21 network and impairs the damage checkpoints those are robustly connected to oncogenic chromosomal abnormalities. PMID- 17416210 TI - Psychosis: atypical limbic epilepsy versus limbic hyperexcitability with onset at puberty? AB - Phencyclidine (PCP), ketamine (Special K), and MK-801 are noncompetitive N-methyl d-aspartate (NMDA) antagonists that produce acute psychosis in humans. The psychosis produced by these psychomimetic drugs is indistinguishable from schizophrenia and includes both positive and negative symptoms. This drug-induced psychosis occurs after puberty in humans. On the basis of the MK-801-induced spike-and-wave activity in rats and increased blood flow and metabolism in brain of patients with psychosis caused by these psychomimetics, this brief review argues that this psychosis is an atypical form of limbic epilepsy. Moreover, there is a specific limbic thalamcortical psychosis circuit that mediates cell injury in limbic cortex of rodents and may mediate this PCP-induced psychosis in humans. It is proposed that this thalamocortical psychosis circuit develops at puberty and can mediate PCP and ketamine-mediated psychosis and possibly the psychosis of schizophrenia, bipolar disease and other disorders that have their onset at puberty. Finally, based on this developmentally regulated psychosis/epilepsy-related thalamocortical circuitry, it is proposed that antiepileptic drugs that promote GABAergic mechanisms may decrease the probability of episodic psychosis from any cause. PMID- 17416212 TI - TASK-like potassium channels and oxygen sensing in the carotid body. AB - Chemosensing by type-1 cells of the carotid body involves a series of events which culminate in the calcium-dependent secretion of neurotransmitter substances which then excite afferent nerves. This response is mediated via membrane depolarisation and voltage-gated calcium entry. Studies utilising isolated cells indicates that the membrane depolarisation in response to hypoxia, and acidosis, appears to be primarily mediated via the inhibition of a background K(+)-current. The pharmacological and biophysical characteristics of these channels suggest that they are probably closely related to the TASK subfamily of tandem-P-domain K(+)-channels. Indeed they show greatest similarity to TASK-1 and -3. In addition to being sensitive to hypoxia and acidosis, the background K(+)-channels of the type-1 cell are also remarkably sensitive to inhibition of mitochondrial energy metabolism. Metabolic poisons are known potent stimulants of the carotid body and cause membrane depolarisation of type-1 cells. In the presence of metabolic inhibitors hypoxic sensitivity is lost suggesting that oxygen sensing may itself be mediated via depression of mitochondrial energy production. Thus these TASK like background channels play a central role in mediating the chemotransduction of several different stimuli within the type-1 cell. The mechanisms by which metabolic/oxygen sensitivity might be conferred upon these channels are briefly discussed. PMID- 17416213 TI - Development of a sensitive liquid chromatography method coupled with a tandem mass spectrometric detection for the clinical analysis of vinflunine and 4-O deacetyl vinflunine in blood, urine and faeces. AB - A sensitive and specific liquid chromatographic method coupled with tandem mass spectrometric detection was set up and fully validated for the simultaneous quantification of vinflunine (VFL) and its pharmacologically active metabolite, 4 O-deacetyl vinflunine (DVFL). The two compounds, as well as vinblastine (used as internal standard), were deproteinised from blood and faeces, analysed on a cyano type column and detected on a Micromass Quattro II system in the positive ion mode after ionisation using an electrospray ion source. In blood, linearity was assessed up to 200 ng/ml for vinflunine and 100 ng/ml for 4-O-deacetyl vinflunine. The lower limit of quantification was validated at 250 pg/ml for both compounds. In other biological media, the linearity was assessed within the same range; the limit of quantification was adjusted according to the expected concentration levels of each compound. This method was first developed in order to identify the structures and to elucidate the metabolic pathway of vinflunine. Thanks to its high sensitivity and specificity, the method has enabled the quantification of vinflunine and 4-O-deacetyl vinflunine in blood at trace levels, and has contributed to the knowledge of vinflunine metabolism by monitoring up to 10 metabolites. PMID- 17416214 TI - High-performance liquid chromatographic method with UV photodiode-array, fluorescence and mass spectrometric detection for simultaneous determination of galantamine and its phase I metabolites in biological samples. AB - Galantamine, an alkaloid isolated from the bulbs and flowers of Caucasian snowdrop (Galanthus woronowii, Amaryllidaceae) and related species, is employed in human medicine for the treatment of various neuromuscular and neurodegenerative diseases. After the administration, the products of oxidative biotransformation (O-desmethyl-galantamine, N-desmethyl-galantamine, galantamine N-oxide) and chiral conversion (epigalantamine) are formed in various concentrations from parent compound. For the identification and determination of galantamine and its phase I metabolites in blood plasma and tissues, a new bioanalytical method based on a reversed-phase high-performance liquid chromatography with UV photodiode-array, fluorescence and mass spectrometric detection was developed, validated and applied to pharmacokinetic and biotransformation studies. Sample preparation included a homogenization of the rat tissues (liver, brain, hypophysis) in a phosphate buffer 0.05 mol/L pH 7.4. Plasma samples and tissue homogenates were purified using a mixed-mode solid phase extraction (Waters Oasis MCX cartridges). Galantamine, its above-mentioned metabolites and the internal standard codeine were separated on a Discovery HS F5 column (Supelco, 150 mmx4.6 mm I.D., 5 microm) at flow rate of 1 mL/min using a linear gradient elution. UV photodiode-array and mass spectrometric detection were employed for the identification of individual galantamine metabolites in various biomatrices, the fluorescence detection (lambdaexcit=280 nm/lambdaemiss=310 nm) was chosen for the quantification of galantamine and its metabolites. The developed method was applicable in liver tissue in the range from 0.50 to 63.47 nmol/g of galantamine, from 0.32 to 41.42 nmol/g of O desmethyl-galantamine, from 0.54 to 69.40 nmol/g of N-desmethyl-galantamine and from 0.70 to 89.03 nmol/g of epigalantamine. Limit of detection was found to be 0.04 nmol/g for galantamine, 0.19 nmol/g for O-desmethyl-galantamine, and 0.07 nmol/g for N-desmethyl-galantamine and epigalantamine. PMID- 17416215 TI - A 4-year treatment with clodronate plus calcium and vitamin D supplements does not improve bone mass in primary biliary cirrhosis. AB - INTRODUCTION: International guidelines for managing osteoporosis in cirrhosis or severe cholestasis indicate a <-2.5 t-score as a cut-off for medical treatment, while no treatment is recommended in the case of osteopenia (t-scores ranging from -1.0 to -2.5). AIM: We conducted a prospective study in primary biliary cirrhosis with a view to optimizing the rationale for the medical treatment of bone loss. METHODS: All naive post-menopausal women with primary biliary cirrhosis were enrolled in the study. Bone metabolism was evaluated by measuring 25-hydroxy-vitamin D, parathyroid hormone, osteocalcin. Bone mineral density was assessed at the lumbar spine by dual-photon X-ray absorptiometry at the baseline and every 2 years for up to 4 years. Patients with either osteopenia or osteoporosis received the following treatment: oral calcium carbonate (1000 mg/day)+vitamin D3 (880 IU/day)+i.m. disodium clodronate 100mg every 10 days for 4 years. RESULTS: Ninety-six patients completed the study: 30 had a normal bone mineral density (group 1), 37 had osteopenia (group 2), 29 had osteoporosis (group 3). No significant differences in biochemical parameters of bone metabolism were observed between the three groups. A total of 288 bone mineral density measurements were taken. Linear regression analysis failed to reveal significant changes in t-score over the follow-up in all groups. CONCLUSIONS: A 4 year treatment with clodronate+calcium/vitamin D3 supplements does not significantly improve osteoporosis or osteopenia in primary biliary cirrhosis women in menopause, but prevents the natural bone loss in these patients. Extensive international trials are warranted to optimize the prevention and treatment of bone loss in primary biliary cirrhosis. PMID- 17416216 TI - A new classification system for retrosternal goitre based on a systematic review of its complications and management. AB - INTRODUCTION: There is no standard definition for thyroid glands extending below the thoracic inlet, and there are no clear guidelines for pre-operatively identifying those patients that may require an intrathoracic approach. We therefore reviewed the current literature in order to establish the current practices regarding the management of retrosternal goitres (RSGs), and propose a classification system to aid pre-operative planning for this important group of patients. MATERIALS AND METHODS: A PubMed Medline search was conducted using the search terms 'retrosternal', 'substernal', 'intrathoracic', 'mediastinal', 'goitre' and 'goiter', resulting in 626 hits. Exclusion criteria reduced the number of papers to the 34 used for this review. RESULTS: A total of 34 papers totaling 2426 patients were included. Eighty-four percent of patients operated on for RSG were achieved via a cervical approach, with the remainder also requiring manubriotomy (3.1%), full sternotomy (6.6%) or thoracotomy (4%). Tracheomalacia occurred in 1% of patients and Superior Vena Cava syndrome (SVC) in 3.2%. There was a clear and highly significant association between the extent and definition of RSG and reported complications, as well as the approach used, with the incidence of tracheomalacia, SVC and need for intrathoracic approach increasing more than 10-fold in cases of RSG reaching the aortic arch. DISCUSSION: There is a clear need to establish a common standard in the definition and description of the extent of RSG. Using our findings, we propose a new, simple, 3-grade classification system of RSGs, based on their relation with the aortic arch and the right atrium. PMID- 17416217 TI - Sample handling strategies for the determination of persistent trace organic contaminants from biota samples. AB - Even after emergence of most advanced instrumental techniques for the final separation, detection, identification and determination of analytes, sample handling continues to play a basic role in environmental analysis of complex matrices. In fact, sample preparation steps are often the bottleneck for combined time and efficiency in many overall analytical procedures. Thus, it is not surprising that, in the last two decades, a lot of effort has been devoted to the development of faster, safer, and more environment friendly techniques for sample extraction and extract clean up, prior to actual instrumental analysis. This article focuses on the state of the art in sample preparation of environmental solid biological samples dedicated to persistent organic pollutants (POPs) analysis. Extraction techniques such as Soxhlet extraction, sonication-assisted extraction, supercritical fluid extraction (SFE), microwave-assisted extraction (MAE), pressurised liquid extraction (PLE) and matrix solid-phase dispersion (MSPD) are reviewed and their most recent applications to the determination of POPs in biota samples are provided. Additionally, classical as well as promising novel extraction/clean-up techniques such as solid phase microextraction (SPME) are also summarized. Finally, emerging trends in sample preparation able to integrate analytes extraction and their adequate clean-up are presented. PMID- 17416218 TI - Optimisation of a matrix solid-phase dispersion method for the determination of organophosphate compounds in dust samples. AB - A fast and inexpensive sample preparation procedure based on the matrix solid phase dispersion (MSPD) technique is proposed for the isolation of several organophosphate esters (mainly employed as flame retardants and plasticizers) from indoor dust samples. Extraction and clean-up were carried out in a single step and target compounds were determined by gas chromatography (GC) with nitrogen-phosphorus detection (NPD). The main parameters affecting extraction yield and selectivity, such as type and amount of dispersant material, clean-up co-sorbent and extraction solvent, were evaluated and optimised. Under final conditions, 0.5 g of dust were dispersed with equal amounts of anhydrous sodium sulphate and Florisil, and loaded on the top of a polypropylene cartridge containing 0.5 g of alumina. The dispersed sample was washed with 2 mL of n hexane to remove the least polar interferences and analytes were eluted with 3 mL of acetone. Recoveries of the proposed method for spiked samples ranged from 80 to 116%, and the day-to-day variability remained between 5 and 10%. Data on levels of organophosphate species in dust from private houses and vehicle cabins are provided. In both cases, the lowest concentrations corresponded to the short chain, non-chlorinated, alkyl organophosphates, whereas mean values above 1 microg g(-1) were measured for the rest of analytes. PMID- 17416219 TI - pH-resistant titania hybrid organic-inorganic sol-gel coating for solid-phase microextraction of polar compounds. AB - A novel titania-hydroxy-terminated silicone oil (titania-OH-TSO) sol-gel coating was developed for solid-phase microextraction of polar compounds. In general, titania-based sol-gel reaction is very fast and need to be decelerated by the use of suitable chelating agents. But in the present work, a judiciously designed sol solution ingredients was used to create the titania-OH-TSO coating without the addition of any chelating agent, which simplified the sol-gel procedure. Thanks to the variety of titania's adsorption sites and their acid-base characteristics, aromatic amines, phenols and polycyclic aromatic hydrocarbons were efficiently extracted and preconcentrated from aqueous samples followed by thermal desorption and GC analysis. The newly developed sol-gel hybrid titania coating demonstrated excellent pH stability, and retained its extraction characteristics intact even after continuous rinsing with a 3 M HCl or NaOH solution for 12 h. Furthermore, it could withstand temperatures as high as 320 degrees C. Practical application was demonstrated through the analysis of six aromatic amines in dye process wastewater. A linearity of four orders of magnitude was obtained with correlation coefficient better than 0.9982. The detection limits ranged from 0.22 to 0.84 microg L(-1) and the repeatability of the measurements was <7.0%. The recoveries of these compounds studied in the wastewater were in the ranges 83.6-101.4%, indicating the method accuracy. PMID- 17416220 TI - Determination of triazine herbicides in sheep liver by microwave-assisted extraction and high performance liquid chromatography. AB - A multi-residue method developed for the analysis of triazine herbicides, simazine, atrazine, propazine and prometryne, in sheep liver is presented. The method is based on microwave-assisted extraction (MAE) of sheep liver using methanol as extractant and analysis of extracts by high performance liquid chromatography (HPLC) and ultraviolet detection. MAE operational parameters, the solvent type and volume, extraction temperature and time, were optimized in detail with respect to extraction efficiency of the target compounds from sheep liver. The recoveries of the method at two different spiked levels were assessed by analyzing spiked liver samples and were found to be in the range from 90 to 102% with good precision (<11%). PMID- 17416221 TI - Determination of molybdenum in environmental samples. AB - Determination of molybdenum in different kinds of environmental samples is often a challenging task for analysts. Its concentration is usually very low and the sample matrix may cause serious interferences during measurement. Therefore, preconcentration and separation methods should be used to solve these problems and render more sensitive, accurate and interference-free determination. Recent developments in sample treatment, such as solid phase and liquid-liquid extraction as well as coprecipitation are presented, including flow-based methodology. In addition, important extension and improvements in analytical methods for determinations of molybdenum are updated. Some examples of speciation analysis are also presented. PMID- 17416222 TI - Improved quality control in gas chromatography interfaced to stable isotope ratio mass spectrometry by application of derivative chromatography. AB - Compound-specific isotope analysis using gas chromatography interfaced to isotope ratio mass spectrometry (GC-IRMS) is a versatile technique for applications ranging from source appointment and the elucidation of biochemical pathways. When delta(13)C values are going to be determined, the sample is combusted to CO(2) and the resulting gas is analyzed relative to a standard with known stable carbon isotope ratio. With the combustion step any information on the identity of a peak is lost. Co-eluting compounds can no more be identified which can lead to significant alterations of the delta(13)C value of the analyte. For improvement of the QA/QC protocols in GC-IRMS, we used first, second, and third order derivative chromatography. The suitability of the technique was studied using mixtures of 2,3,3',4,4',5,5'-heptachloro-1'-methyl-1,2'-bipyrrole (Q1) and 2,2',4,5,5'-pentachlorobiphenyl (PCB 101). By application of different GC oven programs four scenarios ranging from baseline separation to full co-elution were obtained. Derivative chromatography enabled identification of the interference of Q1 with PCB 101 even when both peaks fully co-eluted. Although the delta(13)C values could not be determined from interfered scenarios, the use of derivative spectroscopy will help to prevent acceptance of incorrect data due to co elutions. Derivative chromatography was finally used to study the peak purity of 2,2',3,4,4'-pentabromodiphenyl ether (BDE 85) in technical pentabromo diphenyl ether (DE-71). Already the first order derivative demonstrated that this key-BDE congener was interfered by a compound identified as 2,2',4,4',6,6' hexabromodiphenyl ether (BDE 155). PMID- 17416223 TI - Evaluation of strontium isotope abundance ratios in combination with multi elemental analysis as a possible tool to study the geographical origin of ciders. AB - In order to evaluate alternative analytical methodologies to study the geographical origin of ciders, both multi-elemental analysis and Sr isotope abundance ratios in combination with multivariate statistical analysis were estimated in 67 samples from England, Switzerland, France and two Spanish regions (Asturias and the Basque Country). A methodology for the precise and accurate determination of the (87)Sr/(86)Sr isotope abundance ratio in ciders by multicollector inductively coupled plasma mass spectrometry (MC-ICP-MS) was developed. Major elements (Na, K, Ca and Mg) were measured by ICP-AES and minor and trace elements (Li, Be, B, Al, Sc, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ga, As, Se, Rb, Sr, Y, Mo, Cd, Sn, Sb, Cs, Ba, La, Ce, W, Tl, Pb, Bi, Th and U) were measured by ICP-MS using a collision cell instrument operated in multitune mode. An analysis of variance (ANOVA test) indicated that group means for B, Cr, Fe, Ni, Cu, Se, Cd, Cs, Ce, W, Pb, Bi and U did not show any significant differences at the 95% confidence level, so these elements were rejected for further statistical analysis. Another group of elements (Li, Be, Sc, Co, Ga, Y, Sn, Sb, La, Tl, Th) was removed from the data set because concentrations were close to the limits of detection for many samples. Therefore, the remaining elements (Na, Mg, Al, K, Ca, Ti, V, Mn, Zn, As, Rb, Sr, Mo, Ba) together with (87)Sr/(86)Sr isotope abundance ratio were considered for principal component analysis (PCA) and linear discriminant analysis (LDA). Finally, LDA was able to classify correctly 100% of cider samples coming from different Spanish regions, France, England and Switzerland when considering Na, Mg, Al, K, Ca, Ti, V, Mn, Zn, As, Rb, Sr, Mo, Ba and (87)Sr/(86)Sr isotope abundance ratio as original variables. PMID- 17416224 TI - Combinatorial electrochemistry using metal nanoparticles: from proof-of-concept to practical realisation for bromide detection. AB - Principles and practical application of combinatorial electrochemistry in search for new electroactive materials in electroanalysis have been explored. Nanoparticles of three different metals: silver, gold and palladium have been independently synthesized on the glassy carbon spherical powder surface by electroless deposition process and characterized using both spectroscopic and electrochemical techniques. These three materials were then combined together onto basal plane pyrolytic graphite electrode surface and the application of the combinatorial approach to find the electrode material for bromide detection as model target analyte was demonstrated. The component electroactive for bromide detection was next identified and it was found that silver nanoparticles were the active ones. A composite electrode based on silver nanoparticle modified glassy carbon powder and epoxy resin was then fabricated and it was found to allow accurate determination of bromide. The electroactivity for the bromide determination of the composite electrode was compared with that of a bulk silver electrode and it was shown that the composite electrode is very efficient with a comparable electroactivity with only a portion of precious metals being used for its construction. PMID- 17416225 TI - Development of fast Fourier transform continuous cyclic voltammetry at Au microelectrode in flowing solutions as a novel method for sub-nanomolar monitoring of lidocaine in injection and biological fluids. AB - In this work a novel method for the fast monitoring of lidocaine in flow injection systems has been developed. The fast Fourier transform continuous cyclic voltammetry (FFTCV) at gold microelectrode in flowing solution system was used for determination of lidocaine in its pharmaceutical formulation. The presented technique was very simple, precise, accurate, time saving and economical, compared with all of the previously reported methods. The recommended technique demonstrated some advantages over other reported methods. Firstly, there was no need for the oxygen removal from the test solution. Secondly, a picomolar detection limit was achieved, and additionally, the method was fast enough for the determination of any such compound, in a wide variety of chromatographic methods. The method was linear across the concentration range of 240-1.1 x 10(5) pg mL(-1) (r=0.996) with a limit of detection and quantitation 117.3 and 240 pg mL(-1), respectively. As a conclusion this system offers the requisite accuracy, sensitivity, precision and selectivity to assay lidocaine in injections. PMID- 17416226 TI - Neutral carriers based polymeric membrane electrodes for selective determination of mercury (II). AB - The potentiometric response characteristics of mercury ion-selective membrane electrodes based on 2-amino-6-purinethiol (I(1)) and 5-amino-1, 3, 4-thiadiazole 2-thiol (I(2)) were described. Ion selectivities were tested for various plasticizers, which were used as solvent mediators to incorporate the ionophores into the membrane. Effects of experimental parameters such as membrane composition, nature and amount of plasticizers and additives, pH and concentration of internal solution on the potential response of Hg(2+) electrodes were investigated. The best performance was obtained with the electrode having a membrane composition (w/w) of (I(1)) (3.17%): PVC (31.7%): DOP (dioctylpthalate) (63.4%): NaTPB (sodium tetraphenylborate) (1.58%). The proposed electrode reveals a Nernstian response over Hg(2+) ion in the concentration range of 7.0 x 10(-8) 1.0 x 10(-1) M with limit of detection 4.4 x 10(-8) M. The electrode shows good discrimination toward Hg(2+) ion with respect to most common cations. It shows a short response time (10s) for whole concentration range and can be used for 2 months without any considerable divergence in potentials. For evaluation of the analytical applicability, the electrode was used in the determination of Hg(2+) ion in different environmental and biological samples. The practical utility of the membrane electrode has also been observed in the presence of surfactants. PMID- 17416227 TI - Quartz crystal microbalance biosensor for recombinant human interferon-beta detection based on antisense peptide approach. AB - Quartz crystal microbalance (QCM) biosensors for recombinant human interferon beta (rhIFN-beta) were constructed by utilizing antisense peptides adhering to the QCM gold surfaces. Two antisense peptides, both corresponding to the N terminal fragment 1-14 of rhIFN-beta, were used in this study. Antisense peptide AS-1 was the original antisense peptide and AS-2 was the modified antisense peptide based on the antisense peptide degeneracy. Both antisense peptides were immobilized on the gold electrodes of piezoelectric crystals, respectively, via a self-assembling monolayer of 1,2-ethanedithiol. The binding affinity between rhIFN-beta and each immobilized antisense peptide in solution was evaluated using a quartz crystal microbalance-flow injection analysis (QCM-FIA) system. The dissociation constant of rhIFN-beta on the antisense peptide AS-1 and AS-2 biosensor was (1.89+/-0.101) x 10(-4) and (1.22+/-0.0479) x 10(-5) mol L(-1), respectively. The results suggested that AS-2 had a higher binding affinity to rhIFN-beta than AS-1. The detection for rhIFN-beta using each biosensor was precise and reproducible. The linear response ranges of rhIFN-beta binding to both biosensors were same with a concentration range of 0.12-0.96 mg mL(-1). The results demonstrated the successful construction of highly selective QCM biosensors using antisense peptide approach, and also confirmed the feasibility of increasing antisense peptide binding affinity by appropriate sequence modification. PMID- 17416228 TI - A cam-based laser-induced fluorescence scanner for capillary array electrophoresis. AB - Capillary array electrophoresis (CAE) is an important high throughput analytical technique. Laser-induced fluorescence (LIF) has been the dominant detection method for CAE owing to its low limit of detection (LOD) and wide linear dynamic range (LDR). Linear LIF scanners were first used in CAE because linear motions of an objective match well with a common planar array of capillaries. A problem with linear scanners is that the motor is required accelerating/decelerating so that all capillaries can be properly scanned, which makes motion control complicated and reduces the duty cycle. Rotary scanners were developed to overcome this problem. While rotary scanners have been successfully applied in CAE, the capillaries have to be arranged in a circular format, which can be inconvenient in some cases. In this report, we describe a cam-based LIF scanner as an alternative technique for CAE detection. In this system, a rotary motor is mechanically linked with a capillary holder via a cam. During operation, the motor carries the cam in a rotary motion that drives an array of capillaries on the holder to move back and forth across the objective for fluorescence detection. Using this design, the capillaries can be parallel-arranged in a plane while the motor acceleration/deceleration is avoided. To demonstrate the feasibility of this approach, we constructed a prototype instrument with a constant-velocity scanning distance of approximately 10 mm, a scanning frequency of 3 Hz and a duty cycle of approximately 70%. The scanner exhibited a LOD of 69 pM of fluorescein and a LDR of 3.5 orders of magnitude. Multiplexed capillary SDS PAGE was performed on this scanner for protein separations. PMID- 17416229 TI - Quantification of protein based on single-molecule counting by total internal reflection fluorescence microscopy with adsorption equilibrium. AB - We developed a sensitive single-molecule imaging method for quantification of protein by total internal reflection fluorescence microscopy with adsorption equilibrium. In this method, the adsorption equilibrium of protein was achieved between solution and glass substrate. Then, fluorescence images of protein molecules in a evanescent wave field were taken by a highly sensitive electron multiplying charge coupled device. Finally, the number of fluorescent spots corresponding to the protein molecules in the images was counted. Alexa Fluor 488 labeled goat anti-rat IgG(H+L) was chosen as the model protein. The spot number showed an excellent linear relationship with protein concentration. The concentration linear range was 5.4 x 10(-11) to 8.1 x 10(-10) mol L(-1). PMID- 17416230 TI - Multiplexing fibre optic near infrared (NIR) spectroscopy as an emerging technology to monitor industrial bioprocesses. AB - The application of near infrared spectroscopy in bioprocessing has been limited by its dependence on calibrations derived from single bioreactor at a given time. Here, we propose a multiplexed calibration technique which allows calibrations to be built from multiple bioreactors run in parallel. This gives the flexibility to monitor multiple vessels and facilitates calibration model transfer between bioreactors. Models have been developed for the two key analytes: glucose and lactate using Chinese hamster ovary (CHO) cell lines and using analyte specific information obtained from the feasibility studies. We observe slight model degradation for the multiplexed models in comparison to the conventional (single probe) models, decrease in r(2) values from 89.4% to 88% for glucose whereas for lactate from 92% to 91.8% and a simultaneous increase in the number of factors as the model incorporates the inter-probe variability, nevertheless the models were fit for purpose. The results of this particular application of implementing multiplexed-NIRS to monitor multiple bioreactor vessels are very encouraging, as successful models have been built on-line and validated externally, which proffers the prospect of reducing timelines in monitoring the vessels considerably, and in turn, providing improved control. PMID- 17416231 TI - Simultaneous non-instrumental detection of aflatoxin B1 and ochratoxin A using a clean-up tandem immunoassay column. AB - A set-up and simple method based on the clean-up tandem immunoassay approach was developed for the visual detection of two analytes. The method was based on a 1 mL column with one clean-up layer and two detection immunolayers. As detection immunolayers CNBr-activated Sepharose 4B with coupled secondary rabbit anti-mouse antibodies was used. Different specific antibodies were coupled to each detection immunolayer. The analysis was realised in a competitive ELISA format with visual detection of the developed colour for each detection immunolayer and took 20 min for six sample extracts. The described method was applied to the simultaneous detection of aflatoxin B1 and ochratoxin A in spices with cut-off levels at 5 and 10 microg kg(-1), respectively. Results were confirmed by LC-MS/MS with immunoaffinity column clean-up. PMID- 17416232 TI - Characterisation of thorium-ethylenediaminetetraacetic acid and thorium nitrilotriacetic acid species by electrospray ionisation-mass spectrometry. AB - Electrospray ionisation-mass spectrometry (ESI-MS) has been used to investigate the speciation of Th in the presence of two common organic complexing agents found in radioactive wastes, EDTA and NTA. These ligands may enhance radionuclide migration from nuclear wastes and contaminated land, and characterisation of the complexes formed will improve our understanding of their environmental mobility. When acetic acid and ammonia were used to adjust the pH, the dominant Th-EDTA species changed from the mixed ligand [ThEDTAac](-) complex to [ThEDTAOH](-) and [ThEDTA(OH)(2)ac](3-) as the pH increased, with all species co-existing, to some extent, over the pH range (2.5-10.8). A previously suggested Th-NTA species, [ThNTA(2)](2-), dominates from pH 6.0-8.5 but at high pH (10.0-10.8) NTA was not able to solubilise Th to a measurable extent. In the presence of both EDTA and NTA, Th formed a mixed ligand complex, [ThEDTANTA](3-) which has also been characterised in independent experiments. The identification of the importance of [ThNTA(2)](2-), [ThEDTANTA](3-) and [ThEDTA(OH)(2)](2-) (with an additional acetate ligand) highlights that these species are not included in current speciation databases and thus may impact on predictions of the environmental mobility of Th. ESI-MS has therefore been used to identify and confirm fundamentally important Th complexes. It has proved to be a useful tool for elucidating radionuclide speciation, adding to our knowledge of Th geochemistry and providing a means of critically examining existing stability constant data. PMID- 17416233 TI - Simultaneous determination of tiopronin and d-penicillamine in human urine by liquid chromatography with ultraviolet detection. AB - d-Penicillamine and tiopronin are drugs widely used for the treatment of many diseases. Because of the relatively high frequency of side effects to these compounds, some of which are dose-related, drug monitoring in urine samples during treatment is advisable. In this paper, we describe a simple method for the determination of tiopronin and d-penicillamine in human urine. The method was based on derivatization with 2-chloro-1-methylquinolinium tetrafluoroborate followed by ion-pairing reversed-phase liquid chromatography separation and ultraviolet-absorbance detection. 2-S-quinolinium derivatives of thiols were detected at 355 nm. The derivatization was optimized in terms of pH and time of the reaction. Baseline separation was achieved on an analytical Zorbax SB C-18 (5 microm, 150 mm x 4.6 mm) column with a mobile phase consisting of pH 2.0 0.09 mol L(-1) trichloroacetic acid buffer (component A) and acetonitrile (component B) pumped at 1.0 mL min(-1). Gradient elution was used: 0-4 min, 12% B; 4-8 min, 12 40% B; 8-12 min, 40-12% B. The d-penicillamine and tiopronin standards added to the urine show that the response of the detector is linear within the range studied, from 1 to 200 micromol L(-1) urine. The imprecision ranges for tiopronin and d-penicillamine were within 1.61-8.24% and 2.92-10.60%, respectively. The analytical accuracy for determined compounds was from 97.24 to 109.39%. The lower limits of detection and quantitation were 0.5 micromol L(-1) and 1.0 micromol L( 1) urine, respectively. This method can be used for routine clinical monitoring of the title thiol-drugs. Cysteine can be measured concurrently, if needed. PMID- 17416234 TI - Immunohistochemical examination of epiphyseal growth plates of Japanese Brown cattle with chondrodysplasia. AB - A new type of inherited chondrodysplasia is described in Japanese Brown cattle, but the basic defects of the epiphyseal growth plate (EGP) in the limb long bones, and proliferation and differentiation of the chondrocytes in the EGP, are not yet understood. In the present study, the EGPs of the limb long bones in eight cases of chondrodysplasia and four normal (control) cattle were examined histologically and immunohistochemically. In the control cattle, proliferative chondrocytes (PCs) and hypertrophic chondrocytes (HCs) were arranged in columns parallel to the long axis of the bone, and HCs were situated on the metaphyseal side of the EGP. In all the affected cattle, many chondrocytes with a hypertrophic appearance were detected in the inner areas of the central portion of the EGP. The PC columns were short and arranged irregularly. Bone tissue and small blood vessels were found frequently in these areas. Six affected cattle showed complete EGP-closure. Backscattered electron (BSE) imaging showed that the calcified cartilage matrix was restricted to the lower region of the hypertrophic zone (HZ) of the EGP in the control cattle, while the calcified cartilage matrix and bone tissue were scattered in the inner areas of the EGP in all the chondrodysplastic cattle. Immunohistochemistry revealed type X collagen in the HCs and cartilage matrix of the HZ in the control cattle. In all the affected cattle, type X collagen was detected in apparently hypertrophic chondrocytes in the inner areas of the EGP. Type II collagen was detected in the entire EGP in all the affected cattle, as in the controls. BrdU (5-bromo-2'-deoxyuridine), injected intravenously 1h before euthanasia was detected in many PCs in the EGP in the control cattle; none, however, was detected in the central portion of the EGP in any affected animal. These observations indicate that differentiation into HCs and calcification of cartilage matrix occur in the inner areas of the central portion of the EGP in chondrodysplasia of Japanese Brown cattle. Differentiation into the HCs at this abnormal site may be caused by the inadequate proliferation and disorganization of the PCs. Premature EGP-closure, observed commonly in chondrodysplasia of Japanese Brown cattle, was thought to be caused by replacement of the calcified cartilage in the inner areas of the EGP by bone tissue. PMID- 17416235 TI - Histomorphological and immunohistochemical characterization of equine granulosa cell tumours. AB - Benign (n=33) and malignant metastasizing (n=1) granulosa cell tumours (GCTs) from 34 mares aged 3-21 years, and normal (control) ovaries from nine mares aged 3-10 years, were examined histologically and immunohistochemically (for inhibin alpha, glutathione S-transferase alpha [GSTalpha], c-erbB-2 oncoprotein [cerb], cytokeratin, vimentin, desmin and alpha-actin), the results being related where appropriate to clinical signs and endocrinological data. Availability permitting, serum samples from GCT-affected mares before and several weeks after ovariectomy were examined for the following hormones: oestradiol, progesterone and testosterone (by radioimmunoassay); and inhibin B (by a cross-reactive ELISA). Histological examination revealed that the GCTs were predominantly well differentiated neoplasms. The metastasizing GCT differed immunohistochemically from the benign GCTs in respect of the expression patterns of vimentin, cerb and GSTalpha in the granulosa cells. A notable feature was the presence of Leydig like cells in mares with stallion-like behaviour or elevated serum testosterone, or both. GSTalpha immunolabelling indicated that the Leydig-like cells were potential producers of steroid hormone. From the immunohistochemical and endocrinological findings it was concluded that GCTs produce abnormally high concentrations of inhibin, which reduce the release of follicle-stimulating hormone, leading to atrophy of the contralateral ovary-a finding in 27 of the mares. PMID- 17416236 TI - Expression of Cox-1 and Cox-2 in canine mammary tumours. AB - The aim of this study was to investigate immunohistochemically the expression of cyclooxygenase-1 (Cox-1) and cyclooxygenase-2 (Cox-2) in canine mammary tumours of different histological types. Cox-1 and Cox-2 enzyme expression was evaluated in 70 mammary samples (four normal, six hyperplastic, 60 neoplastic [21 benign and 39 malignant]). Cox-1 expression was identified in all the samples, and Cox-2 in all the mammary lesions except ductal hyperplasia. Two of the four normal mammary gland samples showed focal immunoreactivity for Cox-2. Cox-1 immunoexpression did not differ significantly between benign and malignant lesions (P=0.272). Cox-2 immunoexpression was higher in malignant tumours than in benign counterparts (P<0.001). Of the malignant tumours, carcinosarcomas and tubulopapillary and squamous cell carcinomas had the highest Cox-2 scores. The study showed that malignant tumours had the highest values of Cox-2 expression, and Cox-2 immunolabelling was particularly intense in histological types classically associated with high malignancy. This suggests that nonsteroidal anti inflammatory drugs (NSAIDs), particularly Cox-2 inhibitors, may have a useful role to play in the treatment of canine malignant mammary tumours. PMID- 17416237 TI - Retrospective study of 82 cases of canine lymphoma in Austria based on the Working Formulation and immunophenotyping. AB - In human beings the prevalence of different non-Hodgkin's lymphoma (NHL) subtypes varies according to geographical region. The aim of this study was to classify canine lymphomas in Austria and to compare the results with those of similar studies in other countries. Eighty-two NHLs were classified according to their morphology (based on the Working Formulation) and their immunophenotype (determined with anti-T-cell and anti-B-cell antibodies). Forty-two (51.2%) were of B-cell subtype, 24 (29.3%) of T-cell subtype, and 16 (19.5%) remained unclassified, because of either negative labelling (9/16) or immunoreaction with both antibodies (7/16). Diffuse lymphomas predominated (99%) over follicular lymphomas, while intermediate grade lymphomas (61%) outnumbered high-grade lymphomas (23.2%) and low grade lymphomas (13.4%). The most common subtype was the diffuse large cell lymphoma (40.2%), followed by the large cell immunoblastic lymphoma (13.4%) and the diffuse small lymphocytic lymphoma (13.4%). Follicular large cell lymphoma and small noncleaved cell lymphoma were uncommon (1.2%). Generally, these findings accord with those of similar studies in Western Europe, making the existence of specific risk factors in Austria unlikely. PMID- 17416238 TI - PEPFAR and the fight against HIV/AIDS. PMID- 17416239 TI - Is it time for a public-health approach to eating disorders? PMID- 17416240 TI - The risks of nanotechnology for human health. PMID- 17416241 TI - Darunavir: promising initial results. PMID- 17416242 TI - Pneumococcal vaccination and public health. PMID- 17416243 TI - More evidence to favour newborn screening for cystic fibrosis. PMID- 17416244 TI - Animal research: the debate continues. PMID- 17416245 TI - Fetal alcohol syndrome: law and ethics. PMID- 17416246 TI - Every hospital needs a Gerry Robinson. PMID- 17416247 TI - Clinical update: diagnosis and treatment of essential tremor. PMID- 17416248 TI - Mark Dybul: US Global AIDS Coordinator in charge of PEPFAR. PMID- 17416249 TI - Drug treatments for obesity. PMID- 17416250 TI - Drug treatments for obesity. PMID- 17416251 TI - Drug treatments for obesity. PMID- 17416254 TI - Congenital syphilis in China. PMID- 17416255 TI - Lessons from HIV counselling and testing campaigns. PMID- 17416256 TI - Maternal fish consumption and children's development. PMID- 17416257 TI - A jumpstart for student researchers in Iran. PMID- 17416258 TI - Maternal fish consumption and children's development. PMID- 17416259 TI - Origins of computed tomography. PMID- 17416260 TI - Metals, orthopaedic implants, and risk of cancer. PMID- 17416261 TI - Efficacy and safety of darunavir-ritonavir at week 48 in treatment-experienced patients with HIV-1 infection in POWER 1 and 2: a pooled subgroup analysis of data from two randomised trials. AB - BACKGROUND: The continuing, randomised, multinational, phase IIB POWER 1 and 2 studies aim to evaluate efficacy and safety of darunavir in combination with low dose ritonavir in treatment-experienced HIV-1-infected patients. We did a pooled subgroup analysis to update results at week 48 for patients receiving the recommended dose of darunavir-ritonavir compared with those receiving other protease inhibitors (PIs). METHODS: After 24-week dose-finding phases and primary efficacy analyses, patients randomised to receive darunavir-ritonavir were given 600/100 mg twice daily, and patients receiving control PIs continued on assigned treatment into the longer-term, open-label phase; all patients continued on optimised background regimen. We assessed patients who had reached week 48 or discontinued earlier at the time of analysis; for the darunavir-ritonavir group, only patients who received 600/100 mg twice daily from baseline were included. Analyses were intention-to-treat. The POWER 2 study (TMC114-C202) is registered with ClinicalTrials.gov (NCT00071097). FINDINGS: At week 48, 67 of 110 (61%) darunavir-ritonavir patients compared with 18 of 120 (15%) of control PI patients had viral load reductions of 1 log10 copies per mL or greater from baseline (primary endpoint; difference in response rates 46%, 95% CI 35%-57%, p<0.0001). Based on a logistic regression model including stratification factors (baseline number of primary PI mutations, use of enfuvirtide, baseline viral load) and study as covariates, the difference in response was 50% (odds ratio 11.72, 95% CI 5.75-23.89). In the darunavir-ritonavir group, rates of adverse events were mostly lower than or similar to those in the control group when corrected for treatment exposure. No unexpected safety concerns were identified. INTERPRETATION: Efficacy responses with darunavir-ritonavir 600/100 mg twice daily plus optimised background regimen were greater than those with control PI and were sustained to at least week 48, with favourable safety and tolerability in treatment-experienced patients. This regimen could expand the treatment options available for such patients. PMID- 17416262 TI - Decline in pneumonia admissions after routine childhood immunisation with pneumococcal conjugate vaccine in the USA: a time-series analysis. AB - BACKGROUND: Routine infant immunisation with seven-valent pneumococcal conjugate vaccine (PCV7) began in the USA in 2000. Although invasive pneumococcal disease has declined substantially, the programme's effect on hospital admissions for pneumonia is unknown. We therefore assessed the effect of the programme on rates of all-cause and pneumococcal pneumonia admissions. METHODS: Data from the Nationwide Inpatient Sample, the largest inpatient database available in the USA, were analysed with an interrupted time-series analysis that used pneumonia (all cause and pneumococcal) admission rates as the main outcomes. Monthly admission rates estimated for years after the introduction of PCV7 vaccination (2001-2004) were compared with expected rates calculated from pre-PCV7 years (1997-1999). The year of vaccine introduction (2000) was excluded, and rates of admission for dehydration were assessed for comparison. FINDINGS: At the end of 2004, all-cause pneumonia admission rates had declined by 39% (95% CI 22-52) for children younger than 2 years, who were the target population of the vaccination programme. This annual decline in all-cause pneumonia admissions of 506 (291-675) per 100,000 children younger than 2 years represented about 41,000 pneumonia admissions prevented in 2004. During the 8 study years, 10,659 (2%) children younger than 2 years admitted with pneumonia were coded as having pneumococcal disease; these rates declined by 65% (47-77). This decline represented about 17 fewer admissions per 100,000 children in 2004. Admission rates for dehydration for children younger than 2 years remained stable over the study period. INTERPRETATION: The reduction in all-cause pneumonia admissions in children younger than 2 years provides an estimate of the proportion of childhood pneumonias attributable to Streptococcus pneumoniae in the USA that are vaccine preventable. Our results contribute to the growing body of evidence supporting the beneficial effects of the pneumococcal conjugate vaccines in children. PMID- 17416263 TI - Economic implications of newborn screening for cystic fibrosis: a cost of illness retrospective cohort study. AB - BACKGROUND: Newborn screening for cystic fibrosis might not be introduced if implementation and running costs are perceived as prohibitive. Compared with clinical diagnosis, newborn screening is associated with clinical benefit and reduced treatment needs. We estimate the potential savings in treatment costs attributable to newborn screening. METHODS: Using the UK Cystic Fibrosis Database, we used a prevalence strategy to undertake a cost of illness retrospective snapshot cohort study. We estimated yearly costs of long-term therapies and intravenous antibiotics for 184 patients who were diagnosed as a result of screening as newborn babies, and 950 patients who were clinically diagnosed aged 1-9 years in 2002. Costs of adding cystic fibrosis screening to an established newborn screening service in Scotland were adjusted to 2002 prices and applied to the UK as a whole. Costs were recalculated in US$. FINDINGS: Cost of therapy for patients diagnosed by newborn screening was significantly lower than equivalent therapies for clinically diagnosed patients: mean ($7228 vs $12 008, 95% CI of difference -6736 to -2028, p<0.0001) and median ($352 vs $2442, 1916 to -180, p<0.0001). When we limited the clinically diagnosed group to only those diagnosable with a 31 cystic fibrosis transmembrane regulator mutation assay and assumed similar disease progression in the clinically diagnosed group as in the newborn screening group, we showed that mean ($3,397,344) or median ($947,032) drug cost savings could have offset the estimated cost of adding cystic fibrosis to a UK national newborn screening service ($2,971,551). INTERPRETATION: Including indirect costs savings, newborn screening for cystic fibrosis might have even greater financial benefits to society than our estimate shows. Clinical, social, and now economic evidence suggests that universal newborn screening programmes for cystic fibrosis should be adopted internationally. PMID- 17416264 TI - Plague. AB - Bubonic plague is an often fulminant systemic zoonosis, caused by Yersinia pestis. Conventional microbiology, bacterial population genetics, and genome sequence data, all suggest that Y pestis is a recently evolved clone of the enteric pathogen Yersinia pseudotuberculosis. The genetic basis of this organism's rapid adaptation to its insect vector (the flea) with transmission between mammalian hosts by novel subcutaneous and pneumonic routes of infection is becoming clearer. This transition provides a paradigm for the way in which new pathogens could emerge. Plague in humans is controlled by suppression of rodent reservoir hosts and their fleas and by early detection and treatment of cases of disease. Detection systems for plague in non-endemic regions might now be needed because of a bioterrorism threat. Rapid diagnostic tests are available and a subunit vaccine is in clinical trials. PMID- 17416265 TI - Renin-angiotensin system and cardiovascular risk. AB - The renin-angiotensin system is a major regulatory system of cardiovascular and renal function. Basic research has revealed exciting new aspects, which could lead to novel or modified therapeutic approaches. Renin-angiotensin system blockade exerts potent antiatherosclerotic effects, which are mediated by their antihypertensive, anti-inflammatory, antiproliferative, and oxidative stress lowering properties. Inhibitors of the system-ie, angiotensin converting enzyme inhibitors and angiotensin receptor blockers, are now first-line treatments for hypertensive target organ damage and progressive renal disease. Their effects are greater than expected by their ability to lower blood pressure alone. Angiotensin receptor blockers reduce the frequency of atrial fibrillation and stroke. Renin angiotensin system blockade delays or avoids the onset of type 2 diabetes and prevents cardiovascular and renal events in diabetic patients. Thus, blockade of this system will remain a cornerstone of our strategies to reduce cardiovascular risk. PMID- 17416266 TI - Global perspectives on the sexual and reproductive health of adolescents: patterns, prevention, and potential. AB - Worldwide, societal shifts and behavioural patterns exacerbated by unique developmental vulnerabilities create a confluence of factors that place today's adolescents at heightened risks for poor health outcomes. Country-level data show that continued investment in effective prevention and treatment strategies is essential to protect adolescents' sexual and reproductive health. Whereas strategies must be tailored to the developmental needs of this age group and their social contexts, effective approaches are multifaceted. All adolescents need access to quality youth-friendly services provided by clinicians trained to work with this population. Sex education programmes should offer accurate, comprehensive information while building skills for negotiating sexual behaviours. Girls and boys also need equal access to youth development programmes that connect them with supportive adults and with educational and economic opportunities. Although progress has been made since the 1994 International Conference on Population and Development, adolescents continue to be disproportionately burdened by threats to their sexual and reproductive health. PMID- 17416267 TI - Varicella vaccination in a child with acute lymphoblastic leukaemia. PMID- 17416268 TI - Is there a need for an animal model for spinal manipulation research? PMID- 17416269 TI - Chiropractic care of musculoskeletal disorders in a unique population within Canadian community health centers. AB - OBJECTIVE: This study was part of a larger demonstration project integrating chiropractic care into publicly funded Canadian community health centers. This pre/post study investigated the effectiveness of chiropractic care in reducing pain and disability as well as improving general health status in a unique population of urban, low-income, and multiethnic patients with musculoskeletal (MSK) complaints. METHODS: All patients who presented to one of two community health center-based chiropractic clinics with MSK complaints between August 2004 and December 2005 were recruited to participate in this study. Outcomes were assessed by a general health measure (Short Form-12), a pain scale (VAS), and site-specific disability indexes (Roland-Morris Questionnaire and Neck Disability Index), which were administered before and after a 12-week treatment period. RESULTS: Three hundred twenty-four patients with MSK conditions were recruited into the study, and 259 (80.0%) of them were followed to the study's conclusion. Clinically important and statistically significant positive changes were observed for all outcomes (Short Form-12: physical composite score mean change = 4.9, 95% confidence interval [CI] = 3.8-6.0; VAS: current pain mean change = 2.3, 95% CI = 1.9-2.6; Neck Disability Index: mean change = 6.8, 95% CI = 5.4-8.1; Roland Morris Questionnaire: mean change = 4.3, 95% CI = 3.6-5.1). No adverse events were reported. CONCLUSIONS: Patients of low socioeconomic status face barriers to accessing chiropractic services. This study suggests that chiropractic care reduces pain and disability as well as improves general health status in patients with MSK conditions. Further studies using a more robust methodology are needed to investigate the efficacy and cost-effectiveness of introducing chiropractic care into publicly funded health care facilities. PMID- 17416270 TI - Age- and sex-specific reference values of a test of neck muscle endurance. AB - OBJECTIVE: This study evaluates age- and sex-specific reference values for neck muscle endurance (NME). METHODS: In this cross-sectional study, 116 randomly selected, healthy volunteers (ages 25-64 years) stratified according to age and gender participated. Dorsal and ventral NME was measured in seconds until exhaustion in a laying-down position. A weight of 4 kg for men or 2 kg for women was used in the dorsal procedure. The ventral procedure was performed without external load. Background and physical activity data were obtained and used in the analysis of NME performance. RESULTS: Mean values for dorsal and ventral NME were about 7 and 2.5 minutes for men and 8.5 and 0.5 minutes for women, respectively. The cutoff values for subnormal dorsal and ventral NME were 157 and 56 seconds for men and 173 and 23 seconds for women, respectively. Women's NME was 122% of men's NME in the dorsal (P = .17) and 24% of men's NME in the ventral (P < .0001) procedure. There were no significant differences among age groups. In multiple regression analysis, physical activity explained 4% of variability in the performance of the dorsal NME; and sex explained 37% of the variability in the performance of ventral NME. CONCLUSION: The reference values and the cutoff points obtained could be used in clinical practice to identify patients with a subnormal NME. Sex is an important consideration when using both the test procedure and the reference values. PMID- 17416271 TI - Flexion mobilizations with movement techniques: the immediate effects on range of movement and pain in subjects with low back pain. AB - OBJECTIVE: This study investigates the immediate effects of flexion mobilizations with movement techniques (MWMs) on spinal range of movement in individuals with low back pain and also their impact on pain. A preliminary attempt has been made to describe the clinical profiles of subjects who were thought to benefit from MWMs. METHOD: A small-scale explanatory study was conducted using a crossover design, placebo-controlled, with subjects and assessors blinded. After assessment by physiotherapists, 26 subjects with low back pain with pain on lumbar flexion, thought to be appropriate for treatment with MWMs, participated. Subjects received an MWM intervention and a placebo intervention in a randomized order. Lumbar spinal flexion and extension and pain during flexion were recorded immediately before and after each intervention, using double inclinometry and visual analogue scales. RESULTS: Mean spinal range of movement increased significantly with the MWM intervention, as compared with the placebo (true flexion: MWMs 49.2 degrees [SD 16.4], placebo 45.3 degrees [SD 14.1], P = .005; total flexion: MWMs 76.7 degrees [SD 22.4], placebo 69.7 degrees [SD 21.5], P = .005). Mean pain scores did not change. CONCLUSIONS: The MWMs produced statistically significant, but small, immediate spinal mobility increases but no pain reduction when compared with placebo. By introducing clinical judgment into the subject selection process for the trial, 19 (73%) of 26 subjects benefited from MWMs techniques in terms of range of movement and/or pain intensity, whereas 9 (35%) subjects showed such changes with the placebo intervention. PMID- 17416272 TI - Immediate effects of manipulation of the talocrural joint on stabilometry and baropodometry in patients with ankle sprain. AB - OBJECTIVE: This study assessed the immediate effects of talocrural joint manipulation on stabilometric and baropodometric outcomes in patients with grade II ankle sprain. METHODS: Fifty-two field hockey players (35 men and 17 women) between 18 and 40 years old (mean = 22.5 years, SD = 3.6 years) were included in this study. A simple blind, intrapatient, placebo-controlled, and repeated measures study was carried out. All the patients underwent a baropodometric study performed with a Foot Work force platform (4 times; pre-post placebo group and pre-post intervention group). The sample was subjected to two techniques of manipulative treatment: (a) talocrural joint manipulation and (b) posterior gliding manipulation over the talus. In a second instance, placebo manipulation was applied. Unilateral analysis of variance and multivariate analysis of variance were used for statistical analysis. RESULTS: The results in the intervention group revealed significant differences in the percentage of posterior load on the foot (P = .015) and the percentage of bilateral anterior load (P = .02) before and after the manipulation. The placebo group did not show any change in any of the variables except for area (P = .045). Intergroup comparison revealed statistically significant differences in the increase in percentage of posterior load on the manipulated foot, percentage of bilateral posterior load, percentage of anterior load on the manipulated foot, and percentage of bilateral anterior load (with the exception of the total load on the foot). CONCLUSIONS: The application of caudal talocrural joint manipulation, as compared with placebo manipulation, in athletic patients with grade II ankle sprain redistributed the load supports at the level of the foot. PMID- 17416273 TI - Influence of posture on the range of axial rotation and coupled lateral flexion of the thoracic spine. AB - OBJECTIVE: This study examines the influence of posture on the range of axial rotation of the thorax and the range and direction of the coupled lateral flexion. METHODS: The ranges of mid thoracic axial rotation and coupled lateral flexion were measured in 52 asymptomatic subjects (aged 18-43 years) using an optical motion analysis system. To examine the influence of posture on primary and coupled motion, we initiated axial rotation from a neutral sitting posture and from end-range thoracic flexion and extension. RESULTS: There was a significant decrease in the range of thoracic rotation in flexion compared with the neutral and extended postures (P < .001). The mean range of coupled lateral flexion was 8.9% of the axial rotation range in the neutral posture and increased to 14.3% and 23.2% in the extended and flexed postures, respectively. Patterns of coupled motion varied between subjects, but an ipsilateral pattern was more common in the flexed posture, whereas a contralateral pattern was more common in the neutral and extended postures. CONCLUSIONS: The ranges and patterns of coupled motion of the thorax appear to be strongly influenced by the posture from which the movement is initiated. This has important implications in relation to the interpretation of clinical tests of thoracic motion and in consideration of mechanisms of development of thoracic pain disorders. PMID- 17416274 TI - Should plain films of the lumbar spine be taken in the posterior-to-anterior or anterior-to-posterior position? A study using decision analysis. AB - OBJECTIVE: The objective of this study was to mathematically propose the best position for frontally oriented lumbar spine radiographs. METHODS: Decision analysis with a decision tree was used in this study. Factors used in the analysis were radiation dose, ease of implementation, image quality, cost, and patient comfort. Data on these factors were gathered by reviewing the literature. RESULTS: Radiation dose was found to be the only factor with significant differences between the posterior-to-anterior (PA) position and the anterior-to posterior (AP) position, and PA films gave lower doses to sensitive organs as compared with AP films. There is also some evidence that PA films show better overall image quality as compared with AP films. CONCLUSIONS: Based on the information used in this study, lumbar spine plain film radiographs should be obtained in the PA position. PMID- 17416275 TI - Developmental issues in chiropractic: a South African practitioner and patient perspective. AB - OBJECTIVE: This study explores pertinent aspects of chiropractic practice in contemporary South Africa in terms of the domains of beliefs, philosophy, professional matters, and education. METHODS: Ten practitioners were purposively sampled. From these, 3 were used as gatekeepers to access 6 patients. Semistructured interviews were conducted with the 16 respondents, and computer assisted thematic analysis was used to interpret data. RESULTS: The practitioners in this study distinguish between 2 main views of the chiropractic identity--the "technician" and "physician." The patients interviewed responded that the strengths of the chiropractic profession lie in its social desirability and its health care utility. Two factors seem important in endorsing chiropractic, namely, on the grounds of patient beliefs and philosophical views with respect to health care and the model of practice encountered in the chiropractor's office. However, some patients seem confused by the lack of health care system integration and consequently display uncertainty of the status the chiropractor can claim professionally and educationally. Practitioners portrayed a view, indicating that chiropractic cannot claim coherence in any of the 3 domains investigated. Issues such as the apparent lack of conceptual identity of chiropractic; the absence of a coherent, marketable model of chiropractic practice; and poor interprofessional relationships with mainstream health care stakeholders were observed. The issue of integrated public health care practices as part of the education process and the concomitant perceived lack of exposure especially to black South Africans emerged as interesting and pertinent developmental themes in the local context. CONCLUSIONS: The international discourse related to issues in the domains of philosophy, professionalism, and education and their effect on the practice of chiropractic seem reflected locally. The notion that an increased presence in the public sector might simultaneously increase social awareness and facilitate health care system integration requires further investigation. PMID- 17416276 TI - Chronic mechanical neck pain in adults treated by manual therapy: a systematic review of change scores in randomized clinical trials. AB - OBJECTIVE: This study provides a systematic analysis of group change scores in randomized clinical trials of chronic neck pain not due to whiplash and not including headache or arm pain treated with manual therapy. METHODS: A comprehensive literature search of clinical trials of chronic neck pain treated with manual therapies up to December 2005. Only clinical trials scoring above 11.5 (Amsterdam-Maastricht Scale) were included in the analysis. RESULTS: From 1980 citations, 19 publications were selected. Of the 16 trials analyzed (3 were rejected for poor quality), 9 involved spinal manipulation (12 groups), 5 trials (5 groups) were for spinal mobilization or nonmanipulative manual therapy (1 trial overlapped), and 2 trials (2 groups) involved massage therapy. No trials included trigger point therapy or manual traction of the neck. For manipulation studies, the mean effect size (ES) at 6 weeks for 7 trials (10 groups) was 1.63 (95% confidence interval [CI], 1.13-2.13); 1.56 (95% CI, 0.73-2.39) at 12 weeks for 4 trials (5 groups); 1.22 (95% CI, 0.38-2.06) from 52 to 104 weeks for 2 trials (2 groups). For mobilization studies, 1 trial reported an ES of 2.5 at 6 weeks, 2 trials reported full recovery in 63.8% to 71.7% of subjects at 7 to 52 weeks, and 1 trial reported greater than 2/10 point pain score reduction in 78.3% of subjects at 4 weeks. For massage studies, 1 reported an ES of 0.03 at 6 weeks, whereas the other reported mean change scores of 7.89/100 and 14.4/100 at 1 and 12 weeks of, respectively. CONCLUSIONS: There is moderate- to high-quality evidence that subjects with chronic neck pain not due to whiplash and without arm pain and headaches show clinically important improvements from a course of spinal manipulation or mobilization at 6, 12, and up to 104 weeks posttreatment. The current evidence does not support a similar level of benefit from massage. PMID- 17416277 TI - Sacral fatigue fracture in a female runner: a case report. AB - OBJECTIVE: This case report describes and discusses the clinical presentation, diagnosis, and management of a patient with a sacral fatigue fracture. CLINICAL FEATURES: A 26-year-old female long-distance runner presented with nonspecific low-back and buttock pain that prevented her from training. INTERVENTION AND OUTCOME: Radiographic findings on the patient's lumbar spine and pelvis were interpreted as normal. Single-photon emission computed tomography and magnetic resonance imaging were performed, revealing a fatigue fracture of the left sacral ala. The patient discontinued training for 6 months and gradually returned to running. CONCLUSIONS: A high index of suspicion should prompt investigation with skeletal scintigraphy, computed tomography, or magnetic resonance imaging. Sacral stress fractures may respond well to conservative measures if diagnosed in a timely fashion. PMID- 17416278 TI - Twenty-year-old pathogenic "active" postsurgical scar: a case study of a patient with persistent right lower quadrant pain. AB - OBJECTIVE: This case study describes a patient with persistent right lower quadrant and low back pain who experienced relief after manual mobilization techniques of an old appendectomy scar. CLINICAL FEATURES: A 53-year-old man with pain in the right lower quadrant of the abdomen and low back had previously failed several trials of multimodal treatments. He had an irritated old appendectomy scar in the right lower quadrant. Degenerative disk findings were also noted in the upper lumbar spine. INTERVENTIONS AND OUTCOMES: Manual mobilization of the superficial and deep layers of the scar tissue was applied. The patient experienced an immediate pain reduction after the first treatment. Nine treatments in total were administered to the patient. CONCLUSIONS: Assessment and treatment of "active" scar tissue may comprise an important component of the management of locomotor dysfunction and associated pain syndromes. PMID- 17416279 TI - Introducing the external link model for studying spine fixation and misalignment: part 1--need, rationale, and applications. AB - This is the first article in a series introducing a new animal model, the External Link Model that we propose will allow researchers to produce and study spine lesions with the cardinal biomechanical features of the chiropractic subluxation: fixation (hypomobility) and misalignment. PMID- 17416280 TI - Chiropractic management of cow's milk protein intolerance in infants with sleep dysfunction syndrome: a therapeutic trial. PMID- 17416281 TI - Sulindac sulfone modulates beta-catenin in human cholesteatoma cell culture. AB - BACKGROUND: External auditory canal cholesteatoma (EACC) is a chronic inflammation of the bony ear meatus. Its etiology is not clearly understood. Other than surgical intervention, conservative methods are investigated for different cholesteatomas. Inducing apoptosis seems to be an appropriate strategy. Sulindac sulfone is a new class of targeted and pro-apoptotic drugs. It provokes apoptosis by inducing phosphorylation of beta-catenin, which is a multifunctional protein in the cell-cell adhesion complex. METHODS: EACC-cell cultures were incubated with different concentrations of sulindac sulfone (400 and 800 micromol). After 16, 24, and 48 h, beta-catenin concentrations were determined by ELISA, Western blot, and immunohistochemical analysis. RESULTS: After 48 h incubation with 400 micromol sulindac sulfone, the average level of beta-catenin showed a decrease of 46% (0.004337 microg/mL) from those determined at 16 h with the same concentration of sulindac sulfone. At 800 micromol sulindac sulfone, the treated cell culture showed a reduction of 66.2% (0.003443 microg/mL). Comparing total protein content and the fraction of beta-catenin at different points in time, the concentration of beta-catenin decreased in both EACC cell cultures, 400 micromol (minus 63%) and 800 micromol (minus 81%). CONCLUSIONS: The results presented in this paper are the first to demonstrate the chemopreventive effects of the agent sulindac sulfone on cholesteatomas. The greatest decrease of beta catenin was observed between 16 and 24 h incubation. The inhibitory effect of sulindac sulfone as a local treatment seems to be a useful additional tool for nonsurgical approach to the therapy of EACCs. PMID- 17416282 TI - Transcriptional upregulation of genes related to virulence activation in Entamoeba histolytica. AB - BACKGROUND: To understand the molecular basis of virulence variability in Entamoeba histolytica, this study presents results about differential gene expression induced by E. histolytica trophozoites in liver of hamsters in order to produce experimental amebic liver abscess (ALA) and consequently reactivate its virulence. METHODS: Amebic cultures were studied before (BALA) and after (AALA) inoculation in hamster peritoneal cavity. Markers of pathogenicity such as the rate of erythrophagocytosis, hemolytic activity, and cytotoxic effects on MDCK cell monolayers were evaluated in order to correlate these phenotypic characteristics to differential gene expression between virulent and non-virulent strains. Genotypic variability was determined by genetic polymorphism using the random-amplified polymorphic DNA (RAPD) technique, which defines the parasite genomic plasticity. mRNA differential display was used in order to identify variable transcripts levels. RESULTS: The rate of erythrophagocytosis and hemolytic activity were notably increased in AALA in comparison with BALA E. histolytica cultures, as well as the cytotoxic effect on MDCK cells. An increment in the transcription level of several mRNA was shown. CONCLUSIONS: The RAPD technique allowed us to confirm differences in number and size of polymorphic markers bands between virulent and non-virulent stages, suggesting genomic adaptability in E. histolytica. Eight different genes (membrane-bound acid phosphatase, cysteine proteinase, two different ribosomal proteins, heat shock transcription factor, ribosomal RNA, aldehyde dehydrogenase-1 and patatin-like phospholipase) were sequenced and may be associated with a biological function related to the virulence of E. histolytica. Together these findings show genomic variability between virulent and non-virulent cultures of E. histolytica. PMID- 17416283 TI - Protective effects of erdosteine on doxorubicin-induced hepatotoxicity in rats. AB - BACKGROUND: Oxidative stress has an important role in the pathogenesis of doxorubicin-induced hepatotoxicity. The aim of this study was to investigate the hepatoprotective effects of erdosteine, an antioxidant agent, on doxorubicin induced hepatotoxicity. METHODS: Rats were divided into control, doxorubicin alone (20 mg/kg, i.p.) and doxorubicin plus erdosteine (50 mg/kg/day, oral) groups. At the end of the 10(th) day, liver tissues were removed for light microscopy and analysis. The levels of tissue protein carbonyl content, malondialdehyde and nitric oxide, as well as the activities of superoxide dismutase, catalase, were determined. RESULTS: The tissue of the doxorubicin group showed some histopathological changes such as necrosis, hepatocyte degeneration, sinusoidal dilatation, hemorrhage and vascular congestion and dilatation. In the doxorubicin plus erdosteine group, histopathological evidence of hepatic damage was markedly reduced. Biochemical parameters were consistent with histological parameters. CONCLUSIONS: Doxorubicin caused hepatotoxicity, and erdosteine treatment prevented lipid peroxidation and protein oxidant in liver tissue. PMID- 17416284 TI - Relationship between white blood cell count and Framingham Risk Score in asymptomatic men. AB - BACKGROUND: There is an independent association between white blood cell (WBC) and coronary heart disease (CHD) risk. However, the relationship between WBC and Framingham Risk Score (FRS) remains unclear. METHODS: This is a cross-sectional study on a consecutive sample of 520 white asymptomatic men (mean age 46 +/- 7 years) without CHD. The study population was divided into WBC quartiles (x10(9) cells/L): 1(st) quartile: 3.1-5.3 (n = 139), 2(nd) quartile: 5.4-6.1 (n = 129), 3(rd) quartile: 6.2-7.1 (n = 131), 4(th) quartile: >/=7.2 (n = 121), and into tertiles according to the 10-year FRS: 1(st) tertile (low risk <5%, n = 180, 35%), 2(nd) tertile (intermediate risk 5-12%, n = 210, 40%), 3(rd) tertile (high risk: >/=13%, n = 130, 25%). RESULTS: WBC correlated only weakly with FRS (r = 0.18, p = 0.001). Among individual components of FRS, WBC correlated minimally with smoking (r = 0.12, p = 0.003), systolic blood pressure (r = 0.07, p = 0.1), and high-density lipoprotein cholesterol (r = -0.06, p = 0.1). However, no correlation was observed with age (p = 0.3) and total cholesterol (p = 0.5). Nearly one third (31%) of men in the low-risk (FRS <5%) had WBC count in the 1(st) quartile compared to 20% of those classified as high risk (FRS >/=13%). The prevalence of WBC in the 4(th) quartile increased across FRS tertiles (18, 22, 32%) (p = 0.09). CONCLUSIONS: WBC correlates weakly with FRS or its individual components. Since WBC count is strongly related to CHD, WBC may reflect different components of cardiovascular risk, which might not be captured by traditional cardiovascular risk factors used in calculating FRS. Inflammatory biomarkers afford adjunctive value to FRS and may be used to improve CHD risk stratification. PMID- 17416285 TI - Gastric emptying effect by 4-aminopyridine in patients with chronic spinal cord injury. AB - BACKGROUND: 4-Aminopyridine (4-AP) given to patients with chronic spinal cord injury (SCI) has shown beneficial effects in some somatic and autonomic functions, although patients often develop dyspeptic symptoms. 4-AP is a potassium-channel blocker capable of altering gastro-pyloric functions as demonstrated experimentally. Our objective was to examine the influence of 4-AP treatment on gastric emptying in patients with chronic SCI. METHODS: Gastric emptying was measured by the acetaminophen absorption test in 18 patients (9 with cervical and 9 with thoracic injury), and 9 healthy volunteers. Patients received increasing oral doses, 5 mg day(-1) of oral 4-AP (5-30 mg day(-1)), for 12 weeks. Patients were studied before and at the end of the last week of 4-AP treatment, whereas healthy volunteers (without 4-AP treatment) were studied only once. Whole blood samples of 2.5 mL were drawn at 0 (before 1 g of oral acetaminophen) and at 15, 30, 45, 60, 75, 90, 105 and 120 min postdose. Acetaminophen concentration in plasma was determined by high-pressure liquid chromatography. RESULTS: Treatment with 4-AP significantly delayed stomach emptying in patients with chronic SCI, considering the significant decreasing of acetaminophen absorption (t paired test, p <0.05). This effect did not correlate either to the level or ASIA score of the injury (linear regression correlation analysis, r(2) = 0.003 and 0.015, respectively). No significant differences were observed by comparing data of patients before 4-AP treatment with healthy volunteers. CONCLUSIONS: 4-AP intake in patients with chronic spinal cord injury significantly slowed gastric emptying regardless of level and ASIA score of the injury. PMID- 17416286 TI - Macular edema in unregulated type 2 diabetic patients following glycemic control. AB - BACKGROUND: We undertook this study to evaluate the changes in macular edema of uncontrolled type 2 diabetes mellitus patients with the regulation of hyperglycemia. METHODS: The study population was comprised of 35 type 2 diabetes mellitus patients who had poorly regulated blood glucose values. Ophthalmic examinations including baseline and 6-month macular edema index values of patients by Heidelberg Retinal Tomography (HRT) macular module were done. RESULTS: Twenty four (68.6%) female patients and 11 (31.4%) male patients with a mean age of 50.7 +/- 10.3 (mean +/- SD) years and mean diabetic duration of 9.8 +/- 7.5 years participated in the study. Twenty two (62.9%) did not have diabetic retinopathy (DR), whereas 13 (37.2%) had background DR with macular edema. There was a significant correlation between duration of diabetes and HRT-II macula edema index for the right and left eyes (r = 0.40, p = 0.21 and r = 0.40, p = 0.22, respectively). CONCLUSIONS: Macular edema did not change significantly by regulation of glycemic control in the study group. PMID- 17416287 TI - Effect of paraoxonase 1 polymorphisms on the response of lipids and lipoprotein associated enzymes to treatment with fluvastatin. AB - BACKGROUND: Decreased paraoxonase 1 (PON1) and increased total serum lipoprotein associated phospholipase A(2) (Lp-PLA(2)) activities are suggested to be risk factors for vascular disease. Common PON1 genetic polymorphisms (Q192R and L55M) significantly affect PON1 activity and may also influence high-density lipoprotein (HDL)-associated Lp-PLA(2) activity. However, little is known about the possible effect of PON1 common genetic polymorphisms on the response of lipids as well as PON1 and Lp-PLA(2) activities to treatment with statins. METHODS: Two hundred two hypercholesterolemic patients were treated with fluvastatin 40 mg/day. Fasting serum lipids, Q192R and L55M PON1 polymorphisms as well as PON1 and Lp-PLA(2) (total serum and HDL-associated) activities were determined before and after 6 months of treatment. RESULTS: Fluvastatin treatment did not affect HDL-cholesterol or apolipoprotein (apo) AI but resulted in significant decreases in total cholesterol, triglycerides, low-density lipoprotein-cholesterol, apo B and apo E, as well as total serum Lp-PLA(2) activity. In contrast, PON1 activity significantly increased. None of these changes was influenced by Q192R or L55M PON1 polymorphisms. Overall, HDL-Lp PLA(2) did not change but L55M polymorphism significantly influenced its response to fluvastatin. Specifically, LL homozygotes experienced a significant increase, while M carriers (LM or MM) experienced a non-significant decrease in HDL-Lp PLA(2) activity (p = 0.030 between groups). CONCLUSIONS: Q192R and L55M PON1 polymorphisms did not affect the response of lipids, PON1 and total serum Lp PLA(2) to treatment with a statin. However, L55M PON1 polymorphism significantly modulated the response of HDL-Lp-PLA(2). It should be noted that this is an association study and therefore provides no proof but only indication that PON1 may also exert Lp-PLA(2) activity in HDL. PMID- 17416288 TI - Acute hyperhomocysteinemia induces microvascular and macrovascular endothelial dysfunction. AB - BACKGROUND: Hyperhomocysteinemia (Hhcy) has been shown to induce endothelial dysfunction due to a decrease in bioavailable nitric oxide (NO) by increased vascular oxidant stress. This can be detected as an impairment of endothelium dependent vasodilation in conductance arteries, like brachial or coronary arteries. The effect of Hhcy on endothelial function (EF) in small resistance vessels that critically determine organ perfusion, however, has not been studied systematically in humans. Therefore, we simultaneously determined macro- and microvascular EF in 11 healthy subjects before and during acute Hhcy induced by an oral methionine challenge. METHODS: Macrovascular EF was determined by measuring endothelium-dependent flow-mediated vasodilation of the brachial artery by vascular ultrasound and microvascular EF by measuring skin perfusion during iontophoresis of acetylcholine using laser Doppler fluxmetry. RESULTS: Oral methionine significantly increased homocysteine levels by about 5.1-fold. Acute Hhcy leads to a significant decrease in flow-mediated vasodilation of the brachial artery from 8.1 +/- 0.5% to 3.6 +/- 0.6% and to a significant decrease in the ratio of acetylcholine-stimulated vs. baseline laser Doppler flow in the forearm skin (from 9.2 +/- 1.0- to 7.8 +/- 1.3-fold). CONCLUSIONS: Acute Hhcy impairs macro- as well as microvascular (EF) in humans. PMID- 17416289 TI - 24-hour in-hospital mortality predictions in coronary artery bypass grafting patients. AB - BACKGROUND: The purpose of this study was to determine the factors that can help predict risk of mortality in the first 24 h of coronary artery bypass grafting (CABG), because mortality within a few hours of surgery is a disastrous event for surgeons and the patient's family. METHODS: The study population consisted of 120 in-hospital mortality cases (1.07%) from 11,183 patients who underwent CABG from February 2002 to February 2006 by the same group of surgeons in a referral center. One group consisted of 40/120 (about 33.3%) patients who died during the first 24 h after surgery. The second group consisted of 80/120 patients (66.7%) who died between the 2(nd) and 30(th) day postoperatively. A set of data was gathered from the surgery database of the hospital and analyzed in a univariate model. RESULTS: Among the studied variables, only the following factors proved to be significant: previous percutaneous transluminal coronary angioplasty (PTCA), previous cerebrovascular accident (CVA), cardiopulmonary bypass (CBP) time, and postoperative atrial fibrillation (AF) (p /=35 years, in the countryside of Liaoning, China. Serum lipids levels were proposed by National Cholesterol Education Program Adult Treatment Panel III. RESULTS: Of the study population 34.4% had borderline high total cholesterol (TC), 16.9% had high TC, and 8.8% had low high-density lipoprotein cholesterol (HDL-C). The population with borderline high, high, and very high low-density lipoprotein cholesterol (LDL-C) was 15.7, 3.5, and 0.9%, respectively. In addition, 15.9% had borderline high triglycerides (TG), 17.8% had high TG and 2.0% had very high TG. After adjusting for independent variables, TC and TG had the same risk with fasting plasma glucose (FPG) and body mass index. Risk factors of TC and LDL-C were women, age and FPG. Mongolia nationality as a protective factor positively correlated with TC, LDL-C, and TG. Women, age, drinking, and classification of blood pressure were protective factors but Mongolia nationality was a risk factor for HDL-C. CONCLUSIONS: This study demonstrated that the prevalence of dyslipidemia was relatively high among rural hypertensive patients in China and there were many modifiable risk factors for dyslipidemia such as hypertension, weight, and impaired glucose metabolism. To prevent dyslipidemia and to reduce the increasing burden of cardiovascular disease in rural areas with the undeveloped economy of China, the first aim for us is to reduce these risk factors. PMID- 17416292 TI - Screening for retinopathy of prematurity: results of a 7-year study of underweight newborns. AB - BACKGROUND: Retinopathy of prematurity (ROP) is a serious complication of treatment and extension of survival in premature infants and leads to blindness unless recognized and treated early. An ROP prospective screening survey was performed, enrolling all premature newborns weighing <2000 g at delivery. METHODS: A total of 2014 infants had a retinal evaluation by indirect ophthalmoscopy at 4-6 weeks of age. If any ROP stage was detected, the patient was followed periodically to assess treatment. All premature infants with threshold retinopathy were treated by transscleral cryotherapy at the time of detection. RESULTS: We found 449 infants (22.2%) with ROP in any stage, and 230 (11.42%) with threshold retinopathy who underwent cryotherapy. In the 500-1000 g group (n = 334), 48.2% had ROP in any stage and 92 (27.5%) had threshold retinopathy. In the 1001-1500 g group (n = 1374), 257 (18.7%) had any ROP stage and 122 (8.8%) had stage 3. In the 1501-2000 g group, 306 neonates were evaluated, 31 (10%) had any ROP stage and 16 (5.2%) underwent cryotherapy. A total of 198/230 infants (86%) with threshold retinopathy who received cryotherapy had complete recovery, but 5% developed unilateral and 9% bilateral retinal detachment. There were no complications related to anesthesia. CONCLUSIONS: Overall ROP rate was 2.68/1000 deliveries and 22.2% of premature infants <2000 g had any ROP stage, 11.42% with retinal detachment risk received cryotherapy with 86% successful results. PMID- 17416294 TI - Distribution of HLA-B27 subtypes in ankylosing spondylitis in an Israeli population. AB - BACKGROUND: The aim of this study was to investigate the contribution of the B27 subtypes to ankylosing spondylitis (AS) expression in a group of Jewish patients from Israel and to compare their distribution with that found in Mexican Mestizo patients. Several HLA-B27 alleles have been clearly associated with AS. Among them, B( *)2705 and B( *)2702 are involved in susceptibility in different populations worldwide. The aim of this study was to investigate the associated subtypes in Israel and to compare the results with Mexican Mestizos, who have Semitic genes as part of their ancestry. METHODS: This is a case/control study that included a group of 24 HLA-B27+ Israeli patients with AS and 51 B27+ healthy subjects, most of them Ashkenazi Jews. The B27 subtypes were characterized using a PCR-SSP method. RESULTS: Only B( *)2702 and B( *)2705 alleles were present in AS patients. However, their allele frequency was not significantly different from that found in the control group, probably because of the small sample size: B( *)2702 (patients 62.5% vs. controls 41.2%, OR = 2.31) and B( *)2705 (patients 37.5% vs. controls 50.9%). Two additional alleles were present only in the controls in low frequency: B( *)2707(5.9%) and B( *)2701(1.9%). It is clear that the major susceptibility allele in Ashkenazi Jews from Israel is B( *)2702. CONCLUSIONS: The only allele conferring risk to AS expression in Israeli Jews was B( *)2702, as was previously described in Mexican Mestizos. Populations of Mediterranean ancestry, such as Latin Americans, should be further explored to understand the contribution of ethnicity to the etiopathogenesis of AS. PMID- 17416293 TI - APOC3 promoter polymorphisms C-482T and T-455C are associated with the metabolic syndrome. AB - BACKGROUND: Despite the growing epidemic of the metabolic syndrome (MetS), few studies have evaluated genetic polymorphisms associated with the MetS phenotype. One candidate, APOC3, modulates lipid and lipoprotein metabolism and the promoter polymorphisms C-482T/T-455C are associated with loss of insulin downregulation. METHODS: One hundred twenty two consecutive MetS cases were matched by age, sex and race in a 1:1 case-control design to evaluate the prevalence of common polymorphisms in the following candidate genes: APOC3, APOE, B3AR, FABP2, GNB3, LPL, and PPARalpha and PPARgamma. RESULTS: Compared to controls, MetS subjects exhibited a greater prevalence of APOC3 promoter polymorphisms. Specifically, the frequency of the variant C-482T and T-455C alleles was 70.5 and 81.9% of cases compared to 43.4 and 54.1% in controls, respectively (p <0.0001). Overall, APOC3 promoter variants were associated with a greater likelihood of MetS compared to wild type [C-482T (OR: 4.3; 95% CI: 2.2, 8.6 [p <0.0001]), T-455C (OR: 3.6; 95% CI: 2.0, 6.7 [p <0.0001])]. No material differences were identified between the other genetic variants tested and prevalence of MetS. CONCLUSIONS: These data, therefore, suggest that the APOC3 promoter polymorphisms C-482T and T-455C are associated with the MetS. PMID- 17416295 TI - Antioxidant therapy in idiopathic tinnitus: preliminary outcomes. AB - BACKGROUND: Reactive oxygen species (ROS) play an important role in several pathogenic processes, damaging various structural and functional cellular components. The endothelium is at major risk of radical-induced lesions and this damage is most manifest in microcirculation. It has been recently observed that ROS are implicated in the pathology of the inner ear and the peripheral and central pathways. In a previous study we detected high serum values of ROS in subjects with idiopathic tinnitus. The purpose of the present study was to evaluate the validity of antioxidant treatment in tinnitus sufferers with high ROS values. METHODS: The study considered 31 consecutive patients with unilateral idiopathic tinnitus. The mean pure tone audiometric threshold (PTA), tinnitus loudness, subjective disturbance level [visual analogue scale (VAS) determination], and the indirect ROS dosage 48 h before and after medical treatment were evaluated. Patients underwent an 18-week oral treatment with a mix of phospholipids and vitamins (glycerophosphorylcholine, glycerophosphorylethanolamine, beta-carotene, vitamin C, vitamin E). RESULTS: ROS levels were significantly reduced following antioxidant treatment (malonaldehyde: 2.10 vs. 1.98 mumol/dL, p = 0.003; 4-hydroxynonenal: 2.36 vs. 2.16 mumol/dL, p = 0.002) In addition, great improvement was observed in the reduction of tinnitus (VAS and tinnitus loudness evaluations). No significant changes in audiometric threshold occurred. CONCLUSIONS: Oral antioxidant therapy in patients with idiopathic tinnitus seems to reduce the subjective discomfort and tinnitus intensity and may be considered as an additional treatment modality. PMID- 17416296 TI - HPRT deficiency in a two-month-old child presenting acute renal failure and gout with a new deletion of two bases in exon 3 of the HPRT gene. AB - We describe an HPRT deficiency in a 2-month-old child who presented acute renal failure and gout with normal mental and motor development for age. The patient was diagnosed with Lesch-Nyhan disease and showed a new mutation, a deletion of two bases in exon 3 of the HPRT gene (c.269-270delAT). PMID- 17416297 TI - Hemorheology by microchannel method in males with metabolic syndrome. PMID- 17416299 TI - Hardware flexibility of laboratory automation systems: analysis and new flexible automation architectures. AB - Development of flexible laboratory automation systems has attracted tremendous attention in recent years as biotechnology scientists perform diverse types of protocols and tend to continuously modify them as part of their research. This article is a system level study of hardware flexibility of laboratory automation architectures for high-throughput automation of various sample preparation protocols. Hardware flexibility (system components' adaptability to protocol variations) of automation systems is addressed through the introduction of three main parametric flexibility measures functional, structural, and throughput. A new quantitative measurement method for these parameters in the realm of the Axiomatic Theory is introduced in this article. The method relies on defining probability of success functions for flexibility parameters and calculating their information contents. As flexibility information content decreases, automation system flexibility increases. PMID- 17416300 TI - An automated metrics system to measure and improve the success of laboratory automation implementation. AB - The authors describe a system for collecting usage metrics from widely distributed automation systems. An application that records and stores usage data centrally, calculates run times, and charts the data was developed. Data were collected over 20 months from at least 28 workstations. The application was used to plot bar charts of date versus run time for individual workstations, the automation in a specific laboratory, or automation of a specified type. The authors show that revised user training, redeployment of equipment, and running complimentary processes on one workstation can increase the average number of runs by up to 20-fold and run times by up to 450%. Active monitoring of usage leads to more effective use of automation. Usage data could be used to determine whether purchasing particular automation was a good investment. PMID- 17416301 TI - Simulation-based analysis of fluid flow and electrokinetic phenomena in microfluidic devices. AB - Recent advances in microfabrication techniques, sensing methods, and miniaturization have enabled automated analysis of samples using microfluidic systems. Each unique application requires successful custom development of integrated lab-on-a-chip devices. This involves design, analysis and characterization of individual components, (pumps, valves, mixers, separators, sensors) and the integrated system. In this regard, first-principle-based simulations of the underlying complex multiphysics phenomena can provide detailed understanding of device function. An overview of modeling and simulation-based analysis for the design and development of microfluidic devices is presented. In particular, the authors highlight some key factors affecting the performance of lab-on-a-chip systems such as surface tension effects, analyte dispersion, Joule heating, and mass transport limitations, and delineate the parameters that influence them. The limitations of these modeling techniques and future needs are discussed. PMID- 17416302 TI - Noninvasive fluid property measurements using acoustic methods. AB - The properties of a fluid are normally determined using invasive methods. These methods may lead to possibly contaminating or consuming the sample. When only very small amounts of a valuable sample exist, noninvasive measurement methods are preferred. The properties of fluids can then be used to deduce additional properties based on known relationships. In one case, the surface tension of a fluid may be used to determine the concentration of a fluid. The authors describe a measurement technique involving excitation of the surface of the fluid and the measurement of its response. An acoustic wave is used to both excite and monitor the surface of the liquid. This technique is used to determine the concentration of DMSO and water in solution, and the result determines the amount of fluid needed to deliver an accurate amount of solute in solution. PMID- 17416303 TI - Nanoporous noninvasive cellular electrical activity-based analysis devices. AB - In recent years, rapid advancements have been made in the biomedical applications of microtechnology and nanotechnology. While the focus of such technologies have been primarily on in vitro analytical and diagnostic tools, more recently in vivo therapeutic and sensing applications have gained attention. The long-term integration of cells with inorganic materials provides the basis for novel sensing platforms. The work presented here focuses on the ability to maintain cells long-term in nanoporous silicon-based microenvironments. This article describes the creation of nanoporous, biocompatible, alumina membranes as a platform for incorporation into a cell-based device targeted for in situ recording of cellular electrical activity variations due to the changes associated with the surrounding microenvironments. Studies described herein focus on the interaction of nanoporous alumina substrates embedded in silicon patterned with cells of interest. The fidelity of such a system is demonstrated in terms of viability, proliferation, and functionality. The capability of such microfabricated nanoporous membranes, as in vitro for cell-based assays for sensing and drug delivery applications, is also demonstrated. It has potential in vivo application for therapeutic immunoisolation. PMID- 17416304 TI - A 21st century HPLC workflow for process R&D. AB - A unique high-performance liquid chromatographic (HPLC) workflow specifically designed for the rigors of process development has been developed. A key feature of the workflow is the creation of an HPLC software-hardware platform designed to automatically and systematically screen samples using a matrix of columns and eluents to aggressively search for impurities. The workflow platform was assembled from commercial hardware components and both custom and commercial HPLC software. The platform can be used to challenge existing HPLC methods or to develop new methods. Three real world examples are provided to illustrate the utility of the platform to rigorously assess the complexity of samples and to develop new and improved HPLC methods. PMID- 17416305 TI - The use of a dual dye photometric calibration method to identify possible sample dilution from an automated multichannel liquid-handling system. AB - A significant difference between the reported volumes delivered by a fluid-filled automated liquid-handling system with eight stainless steel fixed tips was observed when a gravimetric method and a dual dye photometric method were used to measure delivered volumes between 20 and 200 microL. A series of experiments, using the default pipetting parameters, was performed which demonstrated that the difference in the reported volume was due to a dilution effect by the system liquid of the liquid handler and did not indicate an error in the delivered volumes. This dilution effect led to a decrease in the reported volume by the dual dye method, which alerted the user to the problem of sample dilution. In contrast, the gravimetric method provided the expected volume, and therefore did not alert the user to the problem. Without optimization of pipetting parameters, the dilution issue can be significant, because the resultant change in the concentration of the compound(s) of interest in the sample could lead to an unidentified error in the sample assay. PMID- 17416306 TI - Verifying liquid-handler performance for complex or nonaqueous reagents: a new approach. AB - Multichannel volume dispensing devices, such as automated liquid handlers, are widely used in drug discovery assays and other high-throughput screening processes. The performance of these systems is heavily based on the ability to deliver proper volumes of specific reagents. Discussed herein is the recent research on broadening existing methods for accurately assessing liquid-handler performance when dispensing complex or nonaqueous reagents. Accurate and reliable adjustment of liquid-handler protocols for varied reagent types could have far reaching adoption in all scientific communities. PMID- 17416307 TI - Assay optimization: a statistical design of experiments approach. AB - With the transition from manual to robotic HTS in the last several years, assay optimization has become a significant bottleneck. Recent advances in robotic liquid handling have made it feasible to reduce assay optimization timelines with the application of statistically designed experiments. When implemented, they can efficiently optimize assays by rapidly identifying significant factors, complex interactions, and nonlinear responses. This article focuses on the use of statistically designed experiments in assay optimization. PMID- 17416308 TI - Automated solutions for total RNA isolation from diverse sample types. AB - In response to the increasing demands to generate larger amounts of quality data faster, specifically in the area of RNA isolation to support gene expression assays, the authors have adopted several automated solutions for isolating total RNA from a variety of sample types, (eg, blood, cells, and tissue). By using automated solutions, the authors were able to increase their throughput without compromising data integrity. PMID- 17416309 TI - Using a microcantilever array for detecting phase transitions and stability of DNA. AB - The authors report the extension of the microcantilever platform to study the thermal phase transition of biomolecules as they are heated. Microcantilever based sensors directly translate changes in Gibbs free energy due to macromolecular interactions into mechanical responses. The authors observed surface stress changes in response to thermal dehybridization of double-stranded DNA oligonucleotides that are attached onto one side of a microcantilever. Once the cantilever is heated, the DNA undergoes a transition as the complementary strand melts, which results in changes in the cantilever deflection. This deflection is due to changes in the electrostatic, ionic, and hydration interaction forces between the remaining immobilized DNA strands. This new technique has allowed the authors to probe DNA melting dynamics and leads to a better understanding of the stability of DNA complexes on surfaces. PMID- 17416310 TI - A low-cost, low-power consumption, miniature laser-induced fluorescence system for DNA detection on a microfluidic device. AB - With a focus on low-cost and low-power consumption, a miniature laser-induced fluorescence (LIF) detection system was assembled using a 635 nm red diode laser as the excitation source and a photodiode element coupled with an operational amplifier for signal collection. The primary elements of the miniature system, namely the laser and the detection system, cost a combined $70 and required only 270 mW of power for operation. When compared to conventional systems assembled using an argon-ion laser source and a photomultiplier tube, this represents a 98% decrease in the cost, and greater than 5000-fold decrease in power consumption. Quantitation of DNA on microdevices using the miniature LIF detection system was also performed with an error of less than 15%. This detection system is a step in the direction of commercializing microfluidic instrumentation by reducing the cost and power required for operation. PMID- 17416311 TI - Automated forensic DNA purification optimized for FTA card punches and identifiler STR-based PCR analysis. AB - Forensic labs globally face the same problem-a growing need to process a greater number and wider variety of samples for DNA analysis. The same forensic lab can be tasked all at once with processing mixed casework samples from crime scenes, convicted offender samples for database entry, and tissue from tsunami victims for identification. Besides flexibility in the robotic system chosen for forensic automation, there is a need, for each sample type, to develop new methodology that is not only faster but also more reliable than past procedures. FTA is a chemical treatment of paper, unique to Whatman Bioscience, and is used for the stabilization and storage of biological samples. Here, the authors describe optimization of the Whatman FTA Purification Kit protocol for use with the AmpFlSTR Identifiler PCR Amplification Kit. PMID- 17416312 TI - Determination of cell colony formation in a high-content screening assay. AB - The use of cell colony formation assays for research and clinical applications to assess the functional integrity of cells after in vitro manipulations is extensive. Key areas include hematopoietic stem cell research, cell transformation studies, and predicting the response of tumors to chemotherapeutic agents. Traditionally, enumeration of colonies has involved laborious and subjective counting by hand using a microscope. Here, laser scanning microplate cytometry has been used to provide an automated high-content readout of the effects of cytostatic agents on colony formation. This approach determines colony number through the application of a volume algorithm. Such an approach permits the differentiation of cytostatic effects where the number of colonies and size remains constant, and cytotoxic effects where the size and number may be reduced. Application of microplate cytometry thus offers significant benefits over alternative analytical methods in the search for novel chemotherapeutic agents. PMID- 17416313 TI - Assessing hERG channel inhibition using PatchXpress. AB - PatchXpress, an automated 16-channel parallel patch clamp system, was used to determine inhibition of human ether-a-go-go related gene (hERG) potassium channels by known blockers. A monoclonal cell line stably expressing hERG potassium channels was generated in CHO-K1 cells. Results were compared to conventional patch clamp experiments using similar voltage protocols and solutions. Success rates were evaluated for cell recordings under a variety of conditions, including Accumax versus trypsin treatment to harvest cells, single versus double compound additions, and polystyrene versus glass-coated compound plates. Finally, polystyrene versus glass-coated compound plates were evaluated, and the authors found that for some compounds (but not all), preparation of compound samples in glass-coated plates resulted in inhibition that more closely matched data obtained by conventional experiments. PMID- 17416314 TI - A fully automated robotic system for high throughput fermentation. AB - High throughput robotic systems have been used since the 1990s to carry out biochemical assays in microtiter plates. However, before the application of such systems in industrial fermentation process development, some important specific demands should be taken into account. These are sufficient oxygen supply, optimal growth temperature, minimized sample evaporation, avoidance of contaminations, and simple but reliable process monitoring. A fully automated solution where all these aspects have been taken into account is presented. PMID- 17416315 TI - A buoyancy-driven compact thermocycler for rapid PCR. AB - The authors have designed a novel convective flow-based thermocycling system capable of performing high-speed DNA amplification via the polymerase chain reaction in a simplified and inexpensive format. Successful amplification of a 191 bp influenza-A target is demonstrated within 25 minutes using a 10 muL reaction volume with no modification to standard laboratory protocols. The system is simple to assemble and can be readily integrated with existing laboratory instrumentation for automated operation. PMID- 17416316 TI - Unusually low plasma concentration of lipopolysaccharide following 160-km race. PMID- 17416318 TI - Establishment of the Australasian paediatric Charcot-Marie-Tooth disease registry. PMID- 17416319 TI - Unilateral lens extraction for high anisometropic myopia in children and adolescents: is this prudent? PMID- 17416320 TI - Probing and intubation as primary treatment for nasolacrimal duct obstruction? PMID- 17416321 TI - Simulated torsional disparity disrupts horizontal fusion and stereopsis. AB - PURPOSE: To investigate how increasing torsional disparity affects clinical measures of the quality of binocular single vision. METHODS: Synoptophore targets were adjusted to present varying degrees of torsional disparity (relative to the fusion-free rest position of the eyes) such as might be observed in a patient with excyclotropia. This disparity was increased in 2 degrees increments up to 10 degrees in 21 normal subjects. The fusional divergence and convergence amplitudes and random-dot stereoacuity were measured at each level of torsional disparity. RESULTS: Horizontal fusional vergence was not significantly altered from the baseline measures at 2 degrees and 4 degrees of torsional disparity. However, both the divergence and the convergence amplitudes declined and were significantly different than the baseline measures at 6 degrees . A similar decline in the random-dot stereoacuity measurements was evident. Fine levels of stereopsis were decidedly affected at greater than 4 degrees of torsional disparity, while more gross levels were similarly affected beyond 6 degrees . CONCLUSIONS: Although human binocular vision can demonstrate the sensory capacity for cyclofusion beyond 10 degrees , torsional disparity of 6 degrees or more significantly degrades horizontal fusional vergence and stereopsis. Torsion in excess of this degree might therefore be a barrier to asymptomatic and comfortable binocular single vision and may require appropriate attention in the planning of more successful surgery for superior oblique palsy and cyclovertical incomitant strabismus. PMID- 17416322 TI - Use of the combined recession and resection of a rectus muscle procedure in the management of incomitant strabismus. AB - BACKGROUND: Scott described a method of achieving the effect of a posterior fixation procedure by combining resection and recession of a rectus muscle, while maintaining the ability to adjust primary position alignment with adjustable sutures. METHODS: A retrospective review of combined recession-resection procedures on rectus muscles between 1998 and 2002 was carried out. RESULTS: We identified 22 patients, 17 females and 5 males. The mean age at presentation was 44 years. The etiology was felt to be paralytic in seven patients (31.8%), mechanical/restrictive in seven patients (31.8%), and due to residual childhood strabismus in eight patients (36.4%). Twenty patients had undergone previous strabismus surgery, ranging from one to six operations (mean, 1.75). Scott procedures were performed on 25 rectus muscles of 22 patients; 12 on the lateral rectus muscles, 7 on the inferior rectus muscles, 3 on the medial rectus muscles, and 3 on the superior rectus muscles. All but one patient had a measurable improvement in gaze incomitance, and 11 of 12 who had a measurement of the field of binocular single vision showed improvement. Twenty patients had follow-up periods of 3 months or more, with a range of 3 to 30 months and a mean of 9.35 months. CONCLUSIONS: The combined recession-resection procedure advocated by Scott has a role in the management of incomitant strabismus. PMID- 17416323 TI - Linear nondisplaced orbital fractures with muscle entrapment. AB - PURPOSE: Linear nondisplaced orbital floor fractures with muscle entrapment occur in the pediatric population. These fractures occur with minimal trauma and few external signs of injury. This study reviews the clinical findings, radiologic findings and interpretations, preoperative and postoperative ocular motility, and outcomes in this subset of orbital fracture patients treated with early surgical repair. METHODS: Review of 12 children with linear orbital floor fractures with inferior rectus muscle entrapment, who underwent surgical repair by a single surgeon. RESULTS: All affected eyes demonstrated significant limitation to elevation and, in seven, depression preoperatively. All 12 patients were operated within 4 days of injury. Radiologist interpretation of computed tomographic (CT) findings recognized fracture in 9 of 11 cases in which a dictated report was available. The radiology report correctly identified muscle entrapment in only three cases and was equivocal in three other cases. The ophthalmologist, based on clinical examination and observation of the CT images, correctly identified findings consistent with linear orbital fracture with muscle entrapment in every case. Surgical findings included a nondisplaced linear floor fracture with muscle entrapment. In the early postoperative period limited elevation was present in 10 patients and limited depression was present in 7. Duction deficits and diplopia resolved in 4 days to 5 months. CONCLUSIONS: Despite prompt surgical repair, limited elevation and depression occur in the early postoperative period, possibly due to muscle edema, hemorrhage, and ischemia. Recovery of normal ocular motility may take weeks or months. External signs of injury may be minimal and radiologic interpretation may not recognize fracture or muscle entrapment. PMID- 17416324 TI - Severe visual impairment in children with mild or moderate retinal residua following regressed threshold retinopathy of prematurity. AB - PURPOSE: To describe clinical features of patients from the Cryotherapy for Retinopathy of Prematurity (CRYO-ROP) trial who, after developing severe ROP in infancy, had minimal or moderate retinal residua in at least one eye but a visual acuity of worse than 20/200 in both eyes at the 10 year examination. METHODS: Data from the 10 year CRYO-ROP Trial follow-up exams were evaluated to identify all patients with retinal outcomes of no retinopathy of prematurity (ROP) residua, straightened temporal vessels, or macular heterotopia in at least one eye, but visual acuity less than 20/200 in both eyes. Presence of optic atrophy, nystagmus, and optic disk cupping and developmental survey results were examined. RESULTS: Of 247 patients examined at 10 years, 16 met our inclusion criteria. At the last age at which the following data were recorded, seven had optic atrophy at the 10 year examination, one had optic disk cupping >0.5 at the 5(1/2) year examination, and eight had nystagmus under binocular conditions at the 24 month examination. Nine patients had a below-normal developmental test score on the Functional Independence Measure for Children (WeeFIM). After clinical data interpretation, we concluded that the predominant cause of visual impairment was postgeniculate disease in five patients, ROP in six patients, and combined anterior and posterior visual pathway disease in two patients; in three patients data were insufficient to make a determination. CONCLUSIONS: Poor visual function with mild to moderate retinal residua of severe ROP in at least one eye is relatively rare. In such patients, anterior, posterior, or combined visual pathway disease can occur. PMID- 17416325 TI - Cataract surgery for congenital cataract: endothelial cell characteristics, corneal thickness, and impact on intraocular pressure. AB - PURPOSE: To investigate whether central corneal thickness (CCT), endothelial cell characteristics, and intraocular pressure (IOP) are affected in patients with previous congenital cataract surgery and to focus on their clinical significance. METHODS: CCT and IOP measurements and specular microscopy were performed in 31 eyes of 17 cases of extracted congenital cataracts and 40 eyes of 20 age- and sex matched participants as control group. The mean of three pachymetry measurements of the central cornea was taken as CCT. IOP was checked using an applanation tonometer. RESULTS: The mean corneal thickness of the eyes with extracted congenital cataract (632+/-45 microm) was significantly greater than that of the control eyes (546+/-33 microm; p<0.001). There was no significant difference in the corneal endothelial cell count, coefficient of variation (CV), and mean cell area (AVG) of endothelial cells between operated eyes and the control group. The mean measured IOP in the operated group (22.8+/-3.3 mm Hg) was significantly greater than IOP in controls (14.1+/-1.8 mm Hg, p<0.001). CONCLUSIONS: Although the corneas were clinically clear and there was no significant difference in endothelial characteristics of eyes with extracted congenital cataract and controls, central corneas of operated eyes were significantly thicker than those of controls. To differentiate the actual glaucoma from ocular hypertension in these patients, the central corneal thickness measurement should strongly be considered. PMID- 17416326 TI - Surgical findings with the tinted AcrySof intraocular lens in children. AB - INTRODUCTION: The AcrySof foldable acrylic intraocular lens (IOL) has become an accepted and frequent means of treating pediatric aphakia. A new version of this lens with blue-light filtering properties purports to offer superior retinal protection. We describe our experience with this tinted lens and compare it to findings with the standard, nontinted IOL. METHODS: A chart review identified 29 eyes of 21 children (tinted lens group; age 11 months to 13 years) who had the tinted IOL implanted. The standard lens group consisted of 38 eyes (31 patients; age 11 months to 15 years). Most eyes had an intact posterior capsule at the time of implantation. RESULTS: The incidence of postoperative inflammation was fairly high in both groups (71% in the tinted lens group, 60% in the standard lens group). The tinted lens group showed a higher incidence of transient inflammation than the standard lens group (p=0.03) but the rates of nontransient sequelae (posterior synechiae, iris synechiae, membranes formation, capsule, or IOL opacification) were similar (66% versus 47%; p=0.11). Posterior capsule opacification occurred in 15 eyes in the tinted lens group and 16 eyes in the standard lens group (p=0.15) but was visually significant in only 10 and 11 eyes, respectively (p=0.77). CONCLUSIONS: Transient inflammation is higher with implantation of tinted versus nontinted IOLs, but long-term inflammatory sequelae are roughly equal, as is the rate of posterior capsule opacification. The question as to whether the chromophore in the tinted lens caused the higher incidence of transient inflammation is unresolved. PMID- 17416327 TI - Incidence of severe retinopathy of prematurity before and after a modest reduction in target oxygen saturation levels. AB - BACKGROUND: Previous studies suggest that reducing target oxygen saturation levels to 85-93% decreases the incidence of severe retinopathy of prematurity (ROP). Our aim was to determine if a more modest reduction in target oxygen saturation levels also reduces ROP incidence. METHODS: One neonatal intensive care unit instituted new oxygen saturation guidelines that changed target levels from the upper 90s to 90-96%. We conducted a retrospective cohort study to determine the proportion of eyes that progressed to (1) threshold or type-1 ROP and (2) stage 3. These proportions were compared between two groups of all eligible infants born up to 10 months before (higher oxygen group, n = 46) and up to 16 months after (lower oxygen group, n = 59) the policy change. Binomial regression was used to calculate relative risks adjusted for birth weight, gestational age, apnea, days of mechanical ventilation, and length of hospital stay. RESULTS: Sixteen of 90 eyes (18%) in the higher oxygen group developed threshold or type-1 ROP versus 16 of 118 eyes (14%) in the lower oxygen group (adjusted relative risk = 0.66, 95% CI = 0.29, 1.51). Twenty-two of 88 eyes (25%) in the higher oxygen group developed stage 3 ROP versus 26 of 118 eyes (22%) in the lower oxygen group (adjusted relative risk = 0.76, 95% CI = 0.43, 1.37). CONCLUSIONS: We observed a small but statistically insignificant reduction in the incidence of severe ROP after a modest reduction in target oxygen saturation levels to 90 to 96% in the first several weeks of life. PMID- 17416328 TI - Outcomes in children with nasolacrimal duct obstruction: Significance of persistent symptoms while stents are in place. AB - To evaluate the correlation between persistent symptoms while stents are in place and final outcome in children with nasolacrimal duct obstruction (NLDO). A retrospective observational case series, with medical record review that included indications for surgery, surgical procedure, presence of symptoms while stents were in place, and final outcome after stent removal. Twenty-eight children with NLDO had nasolacrimal duct stents placed in 42 eyes. Twenty-one of the 42 eyes (50%) had minimal or no signs or symptoms of NLDO while stents were in place, and 18 of 21 (86%) were symptom-free after stent removal. Twenty-one of the 42 eyes (50%) remained symptomatic while stents were in place. Eleven of these 21 eyes (52%) had good outcomes after stent removal. Ten (48%) of these patients had persistent symptoms after stent removal requiring further treatment. The prognosis for a good outcome is excellent if symptoms of NLDO resolve while stents are in place. The prognosis is poorer if symptoms of NLDO persist, but more than half of such patients still have good outcomes. Careful counseling of parents regarding these outcomes should be performed before considering additional interventions. PMID- 17416329 TI - Preexisting posterior capsule defect progressing to white mature cataract. AB - We present two children discovered to have a total cataract in one eye with a posterior subcapsular cataract in the other eye. Sequential photography documented rapid progression of the posterior subcapsular cataract to a preexisting posterior capsule defect and subsequently to a white, mature cataract. We propose that early intervention be considered in cases with any posterior subcapsular changes (no matter how subtle) and history of total cataract in the fellow eye, especially in any situation where loss of follow-up is likely to occur. In the event surgery is not advised, parents should be warned about possible cataract progression and the importance of regular follow-up examinations. PMID- 17416330 TI - Anterior necrotizing scleritis after strabismus surgery in a child. AB - Anterior necrotizing scleritis is a rare but potentially devastating complication of ocular surgery that most often occurs after cataract surgery in elderly patients who may have an underlying systemic autoimmune condition(1) or, less likely, an infectious cause.(2) We describe the management and outcome of a case of bilateral anterior necrotizing scleritis after postoperative infection in a 19 month-old girl who had recently undergone strabismus surgery. PMID- 17416331 TI - Primary superior oblique muscle-levator muscle synkinesis. AB - Congenital ocular aberrant innervations can involve oculomotor, trigeminal, abducens, facial, and glossopharyngeal nerves.(1) Involvement of the trochlear nerve is considered rare or nonexistent.(1) In this report we present a case of upper eyelid retraction while looking in the field of action of the superior oblique muscle, indicating a primary superior oblique-levator muscle synkinesis. PMID- 17416332 TI - Control of intermittent esotropia by head shaking. AB - Head shaking is a rare phenomenon that refers to repetitive, low-amplitude rhythmic oscillations of the head around the cervical axis. In otherwise neurologically normal children with concomitant ophthalmic disease, head shaking (and/or nodding, ie, vertical oscillations) can be an associated finding or a compensatory mechanism to improve visual acuity.(1) Head shaking as a compensatory mechanism to maintain ocular alignment in intermittent esotropia is the subject of this case report. PMID- 17416333 TI - Periosteal fixation in third nerve palsy. PMID- 17416334 TI - The precaruncular approach to the medial orbit. PMID- 17416336 TI - Ligand-binding prediction in the resistance-nodulation-cell division (RND) proteins. AB - The resistance-nodulation-cell division (RND) protein family is a ubiquitous group of proteins primarily present in bacteria. These proteins, involved in the transport of multiple drugs across the cell envelope in bacteria, exhibit broad substrate specificity and act like efflux pumps. In this work, a protein belonging to the RND protein family, AcrB of Escherichia coli was used as a working model to predict in silico the compounds transported by 47 RND proteins. From AcrB we extracted and clustered 14 amino acids directly involved in substrate interactions. This clustering provides enough information to identify 16 groups that correlates with the ligand they extrude, such as proteins expelling aromatic hydrocarbons (SrpB cluster) or proteins expelling heavy metals (CnrA cluster). The relationship between conserved, cluster-specific and variable residues indicates that although the ligand-binding domain is conserved in structure, it has enough flexibility to recognize specifically a diversity of molecules. PMID- 17416337 TI - Integrating subcellular location for improving machine learning models of remote homology detection in eukaryotic organisms. AB - A significant challenge in homology detection is to identify sequences that share a common evolutionary ancestor, despite significant primary sequence divergence. Remote homologs will often have less than 30% sequence identity, yet still retain common structural and functional properties. We demonstrate a novel method for identifying remote homologs using a support vector machine (SVM) classifier trained by fusing sequence similarity scores and subcellular location prediction. SVMs have been shown to perform well in a variety of applications where binary classification of data is the goal. At the same time, data fusion methods have been shown to be highly effective in enhancing discriminative power of data. Combining these two approaches in the application SVM-SimLoc resulted in identification of significantly more remote homologs (p-value<0.006) than using either sequence similarity or subcellular location independently. PMID- 17416338 TI - Tautomeric and microscopic protonation equilibria of some alpha-amino acids. AB - The acid-base equilibria of several amino acids, namely glycine, l-alanine, l valine, l-leucine, l-phenylalanine, l-serine, l-methionine, and l-isoleucine, have been characterized in terms of microscopic protonation constants and tautomeric ratios. In this study, the microscopic protonation constants and tautomeric ratios of eight amino acids in different ethanol-water mixtures have been determined. The variation of microscopic constants of amino acids is discussed on the basis of solute-solvent interactions, and the ratio of dipolar ionic form to neutral form is determined in ethanol-water mixtures (20/80% ethanol by volume). The dipolar ionic form still greatly predominates, even in 80% ethanol, as estimated from potentiometric titration data. PMID- 17416339 TI - Capillary electrophoresis is a sensitive monitor of the hairpin-random coil transition in DNA oligomers. AB - Capillary electrophoresis (CE) has been used to characterize the hairpin-random coil transition of four octamers in the GCxxxxGC minihairpin family, where xxxx is GAAA, TTTC, TTTT, or AAAA. The transition can be monitored by CE because differences in the frictional coefficients of the hairpin and coil forms of each octamer lead to a difference of approximately 9% in the free solution mobilities of the two conformations. The GAAA octamer is unusually stable, with a melting temperature of 65 degrees C. The TTTT octamer forms a minihairpin with a melting temperature of 29 degrees C, the TTTC octamer has a melting temperature of 16 degrees C, and the AAAA octamer has a melting temperature below 0 degrees C. The thermal transitions of the TTTT, TTTC, and AAAA octamers are well fitted by a structure prediction algorithm; however, the GAAA minihairpin is considerably more stable than predicted. The melting temperature of the GAAA minihairpin is reduced to 47 degrees C in aqueous buffers containing 7.2M urea and to 33 degrees C in buffers containing 7.2M urea plus 40% (v/v) formamide. The combined results indicate that CE is a sensitive technique for monitoring conformational transitions in small DNA molecules. PMID- 17416340 TI - Conformation of 3'CMP bound to RNase A using TrNOESY. AB - The conditions for accurately determining distance constraints from TrNOESY data on a small ligand (3'CMP) bound to a small protein (RNase A, <14 kDa) are described. For small proteins, normal TrNOESY conditions of 10:1 ligand:protein or greater can lead to inaccurate structures for the ligand-bound conformation due to the contribution of the free ligand to the TrNOESY signals. By using two ligand:protein ratios (2:1 and 5:1), which give the same distance constraints, a conformation of 3'CMP bound to RNase A was determined (glycosidic torsion angle, chi=-166 degrees ; pseudorotational phase angle, 0 degrees < or = P < or =36 degrees ). Ligand-protein NOESY cross peaks were also observed and used to dock 3'CMP into the binding pocket of the apo-protein (7rsa). After energy minimization, the conformation of the 3'CMP:RNase A complex was similar to the X ray structure (1 rpf) except that a C3'-endo conformation for the ribose ring (rather than C2'-exo conformation) was found in the TrNOESY structure. PMID- 17416341 TI - C5aR-mediated myocardial ischemia/reperfusion injury. AB - The complement system activation can mediate myocardial ischemia and reperfusion (I/R). Inhibition of C5a activity reveals attenuation of I/R-induced myocardial infarct size. However, the contribution of C5a receptor (C5aR) to I/R injury remains to be unknown. Here, we reported that both mRNA and protein for the C5aR were constitutively expressed on cardiomyocytes and upregulated as a function of time after I/R-induced myocardial cell injury in mice. Blockade of C5aR markedly decreased microvascular permeability in ischemic myocardial area and leukocyte adherence to coronary artery endothelium. Importantly, the blocking of C5aR with an anti-C5aR antibody was associated with inhibition in activation of protein kinase C delta (PKC-delta) and induction of PKC-mediated mitogen-activated protein kinase phosphatases-1 (MKP-1) leading to the increased activity of p42/p44 mitogen-activated protein (MAP) kinase cascade. These data provide evidence that C5aR-mediated myocardial cell injury is an important pathogenic factor, and that C5aR blockade may be useful therapeutic targets for the prevention of myocardial I/R injury. PMID- 17416342 TI - The liver X receptor agonist T0901317 acts as androgen receptor antagonist in human prostate cancer cells. AB - T0901317 is a potent non-steroidal synthetic liver X receptor (LXR) agonist. T0901317 blocked androgenic stimulation of the proliferation of androgen dependent LNCaP 104-S cells and androgenic suppression of the proliferation of androgen-independent LNCaP 104-R2 cells, inhibited the transcriptional activation of an androgen-dependent reporter gene by androgen, and suppressed gene and protein expression of prostate specific antigen (PSA), a target gene of androgen receptor (AR) without affecting gene and protein expression of AR. T0901317 also inhibited binding of a radiolabeled androgen to AR, but inhibition was much weaker compared to the effect of the antiandrogens, bicalutamide and hydroxyflutamide. The LXR agonist T0901317, therefore, acts as an antiandrogen in human prostate cancer cells. PMID- 17416343 TI - Demonstration of bile acid transport across the mammalian peroxisomal membrane. AB - It is well established that peroxisomes play a crucial role in de novo bile acid biosynthesis. The primary bile acids resulting from peroxisomal beta-oxidation are conjugated to either glycine or taurine in the peroxisomal lumen by a bile acid aminotransferase (BAT). These conjugated bile acids are subsequently secreted into the bile. In this paper we show that the export of glycine- and taurine-conjugated bile acids from mammalian peroxisomes proceeds via specific transporter. The transport activity of this protein was detected by reconstitution of peroxisomal membrane proteins in liposomes and measuring the uptake of radiolabeled substrates into these proteoliposomes. The transporter was further characterized using this assay, which led to the identification of DIDS as an inhibitor of the peroxisomal bile-acid transporter, and allowed us to establish some kinetic parameters for the transport activity. PMID- 17416344 TI - A possible role for TSLP in inflammatory arthritis. AB - Thymic stromal lymphopoietin (TSLP) is an IL-7-like cytokine that triggers dendritic cell-mediated Th2-type inflammatory responses and is considered as a master switch for allergic inflammation. In this study, we found increased levels of TSLP and, also TNF-alpha as previously reported, in synovial fluid specimens derived from patients with rheumatoid arthritis (RA) when compared with those from patients with osteoarthritis (OA). In addition, TNF-alpha up-regulated TSLP expression in RA- and OA-derived synovial fibroblasts, which was inhibited by IFN gamma. Furthermore, anti-TSLP neutralizing antibody ameliorated a TNF-alpha dependent experimental arthritis induced by anti-type II collagen antibody in mice. Collectively, these results suggest that TSLP, as a downstream molecule of TNF-alpha, may be involved in the pathophysiology of inflammatory arthritis. TSLP might thus play a role not only in allergic diseases but also in inflammatory arthritis such as RA. PMID- 17416345 TI - Induction of cytotoxic T lymphocyte response against Mycobacterial antigen using domain I of anthrax edema factor as antigen delivery system. AB - We have investigated the efficiency of N-terminal 1-260 residues of Edema factor (EFn) as a delivery system for ESAT-6, an antigenic protein of Mycobacterium tuberculosis H(37)R(v), into the cytosol of mammalian cells. The EFn.ESAT-6 recombinant protein was obtained by genetic fusion of EFn and ESAT-6 DNA. Our data shows that in the presence of PA, EFn.ESAT-6 fusion protein is internalized into the cytosol of antigen presenting cells, and the splenocytes produced both Th1 and Th2 cytokines in vitro. Further, EFn.ESAT-6 elicited effective cytotoxic T lymphocyte (CTL) response in an in vitro CTL assay. This study for the first time demonstrates that EFn can be used as a vehicle to deliver heterologous proteins of therapeutic importance. PMID- 17416346 TI - Mutation in the SH1 helix reduces the activation energy of the ATP-induced conformational transition of myosin. AB - The SH1 helix is a joint that links the converter subdomain to the rest of the myosin motor domain. Recently, we showed that a mutation within the SH1 helix in Dictyostelium myosin II (R689H) reduced the elasticity and thermal stability of the protein. To reveal the involvement of the SH1 helix in ATP-dependent conformational changes of the motor domain, we have investigated the effects of the R689H mutation on the conformational changes of the converter, using a GFP based fluorescence resonance energy transfer method. Although the mutation does not seem to strongly affect conformations, we found that it significantly reduced the activation energy required for the ATP-induced conformational transition corresponding to the recovery stroke. Given the effects of the mutation on the mechanical properties of myosin, we propose that the SH1 helix plays an important role in the mechanochemical energy conversion underlying the conformational change of the myosin motor domain. PMID- 17416347 TI - Effects of apoE genotype on macrophage inflammation and heme oxygenase-1 expression. AB - In order to gain a more comprehensive understanding of the aetiology of apolipoprotein E4 genotype-cardiovascular disease (CVD) associations, the impact of the apoE genotype on the macrophage inflammatory response was examined. The murine monocyte-macrophage cell line (RAW 264.7) stably transfected to produce equal amounts of human apoE3 or apoE4 was used. Following LPS stimulation, apoE4 macrophages showed higher and lower concentrations of tumour necrosis factor alpha (pro-inflammatory) and interleukin 10 (anti-inflammatory), respectively, both at mRNA and protein levels. In addition, increased expression of heme oxygenase-1 (a stress-induced anti-inflammatory protein) was observed in the apoE4-cells. Furthermore, in apoE4-macrophages, an enhanced transactivation of the key redox sensitive transcription factor NF-kappaB was shown. Current data indicate that apoE4 macrophages have an altered inflammatory response, which may contribute to the higher CVD risk observed in apoE4 carriers. PMID- 17416348 TI - Gene transfer of cyto-protective molecules in corneal endothelial cells and cultured corneas: analysis of protective effects in vitro and in vivo. AB - The loss of corneal endothelial cells plays a critical role in many corneal diseases and is a common phenomenon following cornea transplantation. In addition, the non-regenerative capacity of human corneal endothelial cells (HCEC) ultimately requires appropriate protection of corneal tissues during ex vivo storage to ensure vitality of the cells. However, only 70% of donor corneas can be used for grafting because of endothelial deficiencies. Corneal endothelial cell loss during storage is mainly induced by apoptotic cell death. This study was undertaken, for proof of principle, to investigate whether over-expression of cyto-protective molecules Bcl-x(L), Bag-1, and HO-1 prevents the loss of corneal endothelial cells both in vitro and in vivo. We demonstrate that gene transfer of both Bcl-x(L) and HO-1 has cyto-protective effects on HCEC in vitro. However, gene transfer of a single cyto-protective molecule does not prevent its rejection upon transplantation in a MHC class I/II disparate rat model. PMID- 17416349 TI - UK114, a YjgF/Yer057p/UK114 family protein highly conserved from bacteria to mammals, is localized in rat liver peroxisomes. AB - Mammalian UK114 belongs to a highly conserved family of proteins with unknown functions. Although it is believed that UK114 is a cytosolic or mitochondrial protein there is no detailed study of its intracellular localization. Using analytical subcellular fractionation, electron microscopic colloidal gold technique, and two-dimensional gel electrophoresis of peroxisomal matrix proteins combined with mass spectrometric analysis we show here that a large portion of UK114 is present in rat liver peroxisomes. The peroxisomal UK114 is a soluble matrix protein and it is not inducible by the peroxisomal proliferator clofibrate. The data predict involvement of UK114 in peroxisomal metabolism. PMID- 17416350 TI - Glucose regulation of CDK7, a putative thiol related gene, in experimental diabetic nephropathy. AB - We previously described the identification of the 3'end of an unknown gene CDK7 using differential display which appeared to be up-regulated in diabetic kidneys [R.A. Page, C.A. Morris, J.D. Williams, C.J. von Ruhland, A.N. Malik, Isolation of diabetes-associated kidney genes using differential display, Biochem. Biophys. Res. Commun. 232 (1997) 49-53]. Here we show that CDK7 is a putative thiol related gene which is regulated by glucose in human and rat renal cells. CDK7 mRNA increased by >threefold in cultured human mesangial cells grown in high glucose for 4 days. In the kidneys of the GK rat, a model of type II diabetes, CDK7 showed a steady age-related increase in mRNA, increasing to >sixfold in 40 week GK rats compared to normoglycemic age-matched Wistar rat kidneys, this increase correlates with progressive hyperglycemia. CDK7 mRNA is widely expressed, showing particularly high levels of expression in rat and human liver, and encodes a putative 338 amino acids highly conserved peptide with several conserved domains, including a cys-pro-arg-cys domain conserved in 15 diverse species which is similar to the catalytic centre of thioredoxin, suggesting a role in oxidative stress. PMID- 17416351 TI - Cis astaxanthin and especially 9-cis astaxanthin exhibits a higher antioxidant activity in vitro compared to the all-trans isomer. AB - In recent years, a number of studies have implicated the potent antioxidant property of astaxanthin in various experimental systems; however, these studies employed only the all-trans isomer. On the other hand, it has been reported that all-trans natural astaxanthin is readily isomerized to cis-trans, especially 9 cis and 13-cis isomers, under certain conditions by chemical analysis; however, the biological activities of the cis isomers of astaxanthin are little known. In the present study, we investigated the antioxidant activity of 9-cis and 13-cis astaxanthin compared to the all-trans isomer in vitro. In a stable radical DPPH scavenging activity test and in rat microsome and rabbit erythrocyte ghost membrane lipid peroxidation systems induced by AAPH and t-BuOOH, respectively, the results apparently showed that cis-astaxanthin, especially 9-cis astaxanthin, exhibited a higher antioxidant effect than the all-trans isomer. In addition, during polyunsaturated fatty acid (PUFA) oxidation, both DHA and linoleic acid hydroperoxides formation were markedly inhibited by astaxanthin isomers addition in the order 9-cis >13-cis >all-trans. Furthermore, 9-cis also exhibited the most effective inhibition of the generation of ROS induced by 6-hydroxydopamine (6 OHDA) in human neuroblastoma SH-SY5Y cells among the astaxanthin isomers, as well as on the degradation of collagen type II induced by DHA and linoleic acid hydroperoxides. The above-mentioned results suggest, for the first time, that cis isomer astaxanthin, especially 9-cis astaxanthin, has a much higher antioxidant potency than that of the all-trans isomer. PMID- 17416352 TI - Structural analysis of four and half LIM protein-2 in dilated cardiomyopathy. AB - Dilated cardiomyopathy (DCM) is a cardiac disease characterized by dilated ventricle and systolic dysfunction. Most of the DCM patients are sporadic cases, but a certain population of DCM patients can be familial cases caused by mutations in genes for sarcomere/Z-disc components including titin/connectin. However, disease-causing mutations could be identified only in a part of the familial DCM patients, suggesting that there should be other disease causing genes for DCM. To explore a novel disease gene for DCM, we searched for mutations in FHL2, encoding for four and half LIM protein 2 (FHL2) in DCM patients, because FHL2 is known to associate with titin/connectin. A missense mutation, Gly48Ser, was identified in a patient with familial DCM. Functional analysis demonstrated that the FHL2 mutation affected the binding to titin/connectin. Because FHL2 protein is known to tether metabolic enzymes to titin/connectin, these observations suggest that the Gly48Ser mutation may be involved in the pathogenesis of DCM via impaired recruitment of metabolic enzymes to the sarcomere. PMID- 17416353 TI - Apolipoprotein-J prevention of fetal cardiac myoblast apoptosis induced by ethanol. AB - Over-consumption of ethanol (EtOH) represents a major health problem. This study was to test the cytotoxicity of EtOH in cardiac stem cells or myoblasts, and the potential protective effect of apolipoprotein-J (ApoJ), a stress-responding, chaperone-like protein in high-density lipoprotein, on EtOH-injured cardiac myoblasts. In culture, EtOH-exposed canine fetal myoblasts underwent apoptosis in a concentration- and time-dependent manner. Expression ApoJ by cDNA transfection markedly reduced EtOH-induced apoptosis in the cells. ApoJ expression also restored partially the mitochondrial membrane potential and prevented the release of cytochrome-c from mitochondria into cytoplasma. Thus, ApoJ serves as a cytoprotective protein that protects cardiac stem cells against EtOH cytotoxicity. PMID- 17416354 TI - Norepinephrine: The redheaded stepchild of Parkinson's disease. AB - Parkinson's disease (PD) affects approximately 1% of the world's aging population. Despite its prevalence and rigorous research in both humans and animal models, the etiology remains unknown. PD is most often characterized by the degeneration of dopamine (DA) neurons in the substantia nigra pars compacta (SNc), and models of PD generally attempt to mimic this deficit. However, PD is a true multisystem disorder marked by a profound but less appreciated loss of cells in the locus coeruleus (LC), which contains the major group of noradrenergic neurons in the brain. Historic and more recent experiments exploring the role of norepinephrine (NE) in PD will be analyzed in this review. First, we examine the evidence that NE is neuroprotective and that LC degeneration sensitizes DA neurons to damage. The second part of this review focuses on the potential contribution of NE loss to the behavioral symptoms associated with PD. We propose that LC loss represents a crucial turning point in PD progression and that pharmacotherapies aimed at restoring NE have important therapeutic potential. PMID- 17416355 TI - Feedback loops in intracellular signal processing and their potential for identifying novel signalling proteins. AB - Mapping of intracellular signalling networks has attracted a lot of attention during recent years. Approaches which have been used to characterise functional and physical relationships within signalling systems and to identify novel components have included bioinformatics, genomics, proteomics and genetics. However, the importance of feedback loops in the regulation of signal processing systems has often been ignored. Here we report a novel focus for our previous screening platform, which evaluates the potential of autocatalytic feedback for functional screening of cDNA expression libraries. We confirm that the use of such systems is feasible. We propose that the use of feedback loops in functional screens may introduce a bias towards the detection of signalling molecules, which are likely to be of key regulatory importance due to the sensitivity of the system towards their expression levels. PMID- 17416356 TI - Autoantibodies against human tropomyosin isoform 5 in ulcerative colitis destroys colonic epithelial cells through antibody and complement-mediated lysis. AB - INTRODUCTION: Patients with ulcerative colitis (UC) have IgG1 antibodies in serum and colon against human tropomyosin isoform 5 (hTM5), a cytoskeletal microfilament protein found intracellularly and on the surface of colonic epithelial cells (EC). These antibodies may be pathogenic in UC. METHODS: Sera from patients with UC (n=110) or Crohn's disease (CD) (n=50) and from healthy individuals (Hl) (n=30) were preincubated with recombinant hTM5 or bovine serum albumin (BSA), then cultured for 4h with (51)Cr-labelled colonic adenocarcinoma cells (LS180). Cytotoxicity was determined by (51)Cr release assay. RESULTS: All serum samples lysed up to 36% of LS180 cells regardless of the source of the serum. However, adding hTM5 to UC, but not to CD or HI, sera reduced cytotoxicity by up to 75%. This hTM5-induced inhibition of cytotoxicity was found especially with sera from UC patients with active disease, and was found even after total colectomy. The hTM5-induced inhibition was mediated by purified IgG from UC sera. Complement was involved since hTM5-induced inhibition of cytotoxicity declined with either heat inactivation of the sera or premixing sera with Fc fragments. CONCLUSIONS: This study shows that hTM5-specific IgG autoantibodies present in UC sera destroy LS180 cells by antibody and complement-mediated lysis. Such a phenomenon was not seen in CD or HI. This suggests an autoantigenic role of hTM5 and anti-hTM5 antibodies in the pathogenesis of UC. This observation may lead to novel diagnostic and therapeutic possibilities. PMID- 17416357 TI - An enhanced agent based model of the immune system response. AB - We describe some recent enhancements introduced in C-ImmSim, a simulator of the immune system response that we have been developing for a number of years along with preliminary results produced by the simulation of the Highly Active Anti Retroviral Therapy in HIV-1 infected patients. PMID- 17416358 TI - B-spline interpolation for bend intra-oral radiographs. AB - Digital subtraction radiography (DSR) has been demonstrated to improve the detection of minute bone changes during the diagnosis of Periodontal diseases. However, during a dental X-ray session, it is not possible to control the bending of the X-ray film when installed in a patient's mouth. This leads to errors in the subsequent analysis. Error compensation can be done by a warping transformation, in conjunction with a reference wire grid attached to the X-ray film. However, the wire grid has to be attached to the film manually. The grid itself is a source of error in that it obscures the image and causes information loss. In this paper we propose a compensation method without the use of a reference grid. It is a simple algorithm based on B-spline interpolation and image scaling. No prior information about the bending is required. When compared with warping transformation, the method induces less disturbance to the pixel values during the compensation process. As a result, the proposed method produces more accurate image data for subsequent analysis. PMID- 17416359 TI - Mutagenic effect of freezing on mitochondrial DNA of Saccharomyces cerevisiae. AB - Although suggested in some studies, the mutagenic effect of freezing has not been proved by induction and isolation of mutants. Using a well-defined genetic model, we supply in this communication evidence for the mutagenic effect of freezing on mitochondrial DNA (mtDNA) of the yeast Saccharomyces cerevisiae. The cooling for 2 h at +4 degrees C, followed by freezing for 1 h at -10 degrees C and 16 h at 20 degrees C resulted in induction of respiratory mutations. The immediate freezing in liquid nitrogen was without mutagenic effect. The study of the stepwise procedure showed that the induction of respiratory mutants takes place during the freezing at -10 and -20 degrees C of cells pre-cooled at +4 degrees C. The genetic crosses of freeze-induced mutants evidenced their mitochondrial rho- origin. The freeze-induced rho- mutants are most likely free of simultaneous nuclear mutations. The extracellular presence of cryoprotectants did not prevent the mutagenic effect of freezing while accumulation of cryoprotectors inside cells completely escaped mtDNA from cryodamage. Although the results obtained favor the notion that the mutagenic effect of freezing on yeast mtDNA is due to formation and growth of intracellular ice crystals, other reasons, such as impairment of mtDNA replication or elevated levels of ROS production are discussed as possible explanations of the mutagenic effect of freezing. It is concluded that: (i) freezing can be used as a method for isolation of mitochondrial mutants in S. cerevisiae and (ii) given the substantial development in cryopreservation of cells and tissues, special precautions should be made to avoid mtDNA damage during the cryopreservation procedures. PMID- 17416360 TI - Plasmodium yoelii: contribution of oocysts melanization to natural refractoriness in Anopheles dirus. AB - It is well known that Anopheles dirus is naturally refractory to rodent malaria parasite, Plasmodium yoelii, but the mechanism is still largely unknown. Here, we found that some P. yoelii taken into An. dirus could develop into oocysts, but oocysts were partially melanized at 7 days and completely melanized at 15 days post-infectious blood meal. Transmission electronic microscopy could find the melanized P. yoelii oocysts in An. dirus as early as 5 days post-infection, with a few haemocytes attaching to the melanized oocysts, indicating a typical humoral melanization reaction. Although the change of protein pattern at 24h post infection suggested that other unknown mechanisms and/or factors might be involved in killing ookinetes, our data implied that oocysts melanization was one of the mechanisms of An. dirus to block P. yoelii development. In addition, activity of phenoloxidase, such as monophenol oxidase and o-diphenoloxidase, in haemolymph of An. dirus fed on infectious blood meal was much higher than that of mosquitoes fed on 5% glucose or normal mouse blood (p<0.05), implying the possible role of PO in oocysts melanization by An. dirus. PMID- 17416361 TI - Identification and characterization of a novel toxin-antitoxin module from Bacillus anthracis. AB - Comparative genome analysis of Bacillus anthracis revealed a pair of linked genes encoding pemK (K, killer protein) and pemI (I, inhibitory protein) homologous to pem loci of other organisms. Expression of PemK in Escherichia coli and Bacillus anthracis was bacteriostatic whereas the concomitant expression of PemI reversed the growth arrest. PemK expression effectively inhibited protein synthesis with no significant effect on DNA replication. Coexpression and interaction of these proteins confirmed it to be a Type II addiction module. Thermal denaturation analysis reflected poor conformational stability of PemI as compared to PemK. Circular dichroism analysis indicated that PemI contains twice the amount of beta sheets as PemK. Gel retardation assays demonstrated that PemI binds to its upstream DNA sequence. This study reports the first evidence of an active chromosome encoded toxin-antitoxin locus in B. anthracis. PMID- 17416362 TI - Modulation of nuclear internalization of Tat peptides by fluorescent dyes and receptor-avid peptides. AB - The nuclear internalization of biomolecules by Tat peptide provides a mechanism to deliver drugs to cells. However, translocation of molecular imaging probes to the nucleus may induce undesirable mutagenesis. To assess the feasibility of retaining its cell permeating effect without nuclear translocation, Tat-peptide was conjugated with a somatostatin receptor (STR)-avid ligand (Oct) and labeled with fluorescent dyes. The results show that Tat-Oct-5-FAM (fluorescein 5' carboxylic acid) remained in the cytoplasm of STR-positive AR42J cells. Co incubation of Tat-Oct-5-FAM with ATP induced nuclear translocation. These data suggest that both dye and Oct-STR endocytosis complex could modulate nuclear internalization of Tat peptides. PMID- 17416363 TI - Preferences of subfertile women regarding elective single embryo transfer: additional in vitro fertilization cycles are acceptable, lower pregnancy rates are not. AB - With identical pregnancy rates after elective single embryo transfer (ET) and double ET strategies consisting of three cycles of IVF or intracytoplasmic sperm injection (ICSI) plus transfers of thawed/frozen embryos if available, 46% of the women undergoing IVF/ICSI favor elective single ET. If elective single ET lowers pregnancy chances with 1%, 3%, or 5%, the percentage of women preferring elective single ET drops to 34%, 24%, and 15%, respectively. If four, five, or six cycles with elective single ET are needed to match the success rate of three cycles with double ET, the percentage of women with a preference for elective single ET drops from 46% to 40%, 36%, and 35% respectively. PMID- 17416364 TI - Effect of tamoxifen treatment on the semen quality and fertility of the male rat. AB - OBJECTIVES: To determine the effects of tamoxifen treatment on the seminal quality and fertility of the male Wistar rat. DESIGN: Experimental prospective study. SETTING: Animal research and university laboratory facility. SUBJECT(S): Seventy-five-day-old male and female Wistar rats. INTERVENTION(S): Sperm quality parameters were assayed in seminal and epididymal sperm samples in control and treated rats at a dose of 0.4 mg tamoxifen/kg per day. In addition, mating studies were performed, and different fertility parameters were assayed. RESULT(S): Tamoxifen treatment significantly decreased sperm concentration and motility in seminal and epididymal sperm. Sperm viability and hypo-osmotic swelling test results were shown not to be altered. The copulatory plug was absent or severely impaired in tamoxifen-treated males. When mating experiments were performed, a significant decrease in the fertility index and increased percentages of preimplantation and postimplantation embryo loss were also observed. CONCLUSION(S): Tamoxifen treatment significantly altered sperm quality in seminal and epididymal sperm. These alterations were present in testis and epididymis, and additional negative effects on the sexual accessory glands were observed. Finally, these alterations were capable of seriously compromising fertility ability of these male rats. PMID- 17416365 TI - AZFc partial deletions in Chilean men with severe spermatogenic failure. AB - OBJECTIVE: To determine the prevalence of AZFc subdeletions in infertile Chilean men with severe spermatogenic impairment. DESIGN: Prospective analysis. SETTING: University infertility clinic. PATIENT(S): Ninety-five secretory azo/oligozoospermic men without AZFc Y chromosome microdeletions: 71 whose testicular histology showed severe spermatogenic impairment and 24 who exhibited reduced testicular volume and elevated serum FSH levels. As controls, we studied 77 men (50 fertile and/or normozoospermic, and 27 with azoospermia and normal spermatogenesis). INTERVENTION(S): Peripheral blood was drawn to obtain genomic DNA for polymerase chain reaction (PCR) digestion assays of DAZ-sequence nucleotide variants and for AZFc-STS PCR after a complete testicular characterization (biopsy, hormonal, and physical evaluation). MAIN OUTCOME MEASURE(S): DAZ genes and AZFc subdeletion types. RESULT(S): In cases we observed two "gr/gr" subdeletions (2.1%), one with absence of DAZ1/DAZ2 (g1/g2 subtype), and the other with absence of DAZ3/DAZ4 (r2/r4 subtype). Additionally, we found a g1/g3 subdeletion in a patient with Sertoli-cell-only syndrome. In controls, we observed two gr/gr subdeletions with absence of DAZ1/DAZ2 (2.6%) in a fertile/normozoospermic and in an obstructive azoospermic man. CONCLUSION(S): AZFc subdeletions do not seem to cause severe impairment of spermatogenesis. Moreover, gr/gr-DAZ1/DAZ2 subdeletions do not appear to affect fertility in Chilean men. PMID- 17416366 TI - Empowering patients undergoing in vitro fertilization by providing Internet access to medical data. AB - OBJECTIVE: To study the effect of an Internet-based personal health record on the empowerment of patients undergoing IVF. DESIGN: Randomized clinical trial. SETTING: Patients undergoing IVF and intracytoplasmic sperm injection (ICSI) in an academic research environment. PATIENT(S): We selected patients who were undergoing an IVF or ICSI treatment, have an Internet connection, and speak fluent Dutch. INTERVENTION(S): An Internet-based personal health record that provides patients with general and personal information concerning their given treatment and that also provides facilities for communication with fellow patients and physicians. MAIN OUTCOME MEASURE(S): Patient empowerment (measured as a multidimensional concept consisting of self-efficacy, actual and perceived knowledge, and involvement in the decision process), patient satisfaction, meaning of infertility problems, social support, anxiety, and depression. RESULT(S): A total of 91 female and 89 male participants were suitable for analysis. No significant differences were observed in per person change in patient empowerment. We did not find any significant differences regarding per person change in patient satisfaction, the meaning of infertility problems, social support, anxiety, and depression. CONCLUSION(S): Usage of the personal health record did not have any effects on patient empowerment, but, at the same time, the study did not find that the personal health record had any significant adverse effects either. PMID- 17416367 TI - Three-dimensional power Doppler ultrasound diagnosis and conservative treatment of ectopic pregnancy in a cesarean section scar. AB - OBJECTIVE: To describe three-dimensional ultrasonographic features of an ectopic pregnancy in a cesarean scar. DESIGN: Description of a case. SETTING: Case report of one patient. PATIENT(S): A 38-year-old women with three previous cesarean deliveries. INTERVENTION(S): Three-dimensional ultrasonography was performed for diagnosis and treatment. MAIN OUTCOME MEASURE(S): Local administration of methotrexate under ultrasonographic guidance. RESULT(S): Ultrasonographic study revealed a gestational sac in the anterior wall of the uterine isthmus with peritrophoblastic flow. It was treated conservatively and successfully with local methotrexate administration under ultrasonographic guidance. CONCLUSION(S): The early diagnosis of ectopic cesarean scar pregnancy allows the conservative treatment with local administration of methotrexate under ultrasonographic guidance. PMID- 17416368 TI - Seasonal reproductive biology of two species of freshwater catfish from the Venezuelan floodplains. AB - Oxydoras sifontesi and Pimelodus blochii are seasonal breeder fish. Spawn occurs once a year over a short interval of time, at the beginning of the rainy season. The gonadosomatic index (GSI), and plasma levels of steroid hormones of P. blochii and O. sifontesi were studied from fish samples, collected from the Portuguesa River (Portuguesa State, Venezuela) in 1998 and 2004-2005, respectively. Gonadal tissue samples were obtained and processed for histology. A macroscopic classification of the degree of gonadal maturation was performed using a six-stage maturity scale. Data was analyzed and compared as a function of the gonadal maturation stage. The GSI of both O. sifontesi and P. blochii increases from stages II to V (preparatory and prespawning periods) and decreases in the stage VI (postspawning). In males, the GSI was usually lower than in females. In both species, the higher plasmatic concentration of 17beta-estradiol (17beta-E) and testosterone (T) were obtained from specimens in stages IV or V. A significant decrease in both hormones was observed in stage VI, except for the males of P. blochii where T concentration did not change between gonadal stages IV-VI. The relevance of these results is discussed in relation to the seasonality and the particular hydrological conditions of the region. PMID- 17416369 TI - Multivariate economic performance assessment of an MPC controlled electric arc furnace. AB - Economic performance is very important to advanced process control projects investigating whether the investment of control technology is worthwhile. In this paper economic performance assessment of a simulated electric arc furnace is conducted. The dependence of controlled variables and the corresponding economic impact are highlighted. PMID- 17416370 TI - Dose-dependent effect of rosuvastatin on apolipoprotein B-100 kinetics in the metabolic syndrome. AB - In a randomized, double-blind, crossover trial of 5-week treatment period with placebo or rosuvastatin (10 or 40 mg/day) with 2-week placebo wash-outs between treatments, the dose-dependent effect of rosuvastatin on apolipoprotein (apo) B 100 kinetics in metabolic syndrome subjects were studied. Compared with placebo, there was a significant dose-dependent decrease with rosuvastatin in plasma cholesterol, triglycerides, LDL cholesterol, apoB and apoC-III concentrations and in the apoB/apoA-I ratio, lathosterol:cholesterol ratio, HDL cholesterol concentration and campesterol:cholesterol ratio also increased significantly. Rosuvastatin significantly increased the fractional catabolic rates (FCR) of very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL) and LDL apoB and decreased the corresponding pool sizes, with evidence of a dose-related effect. LDL apoB production rate (PR) fell significantly with rosuvastatin 40 mg/day with no change in VLDL and IDL-apoB PR. Changes in triglycerides were significantly correlated with changes in VLDL apoB FCR and apoC-III concentration, and changes in lathosterol:cholesterol ratio were correlated with changes in LDL apoB FCR, the associations being more significant with the higher dose of rosuvastatin. In the metabolic syndrome, rosuvastatin decreases the plasma concentration of apoB-containing lipoproteins by a dose-dependent mechanism that increases their rates of catabolism. Higher dose rosuvastatin may also decrease LDL apoB production. The findings provide a dose-related mechanism for the benefits of rosuvastatin on cardiovascular disease in the metabolic syndrome. PMID- 17416371 TI - Uric acid level and its association with carotid intima-media thickness in patients with hypertension. AB - OBJECTIVE: Carotid intima-media thickness (C-IMT) measured noninvasively by ultrasonography is now widely used as a surrogate marker for atherosclerotic disease and directly associated with increased risk of cardiovascular disease. Hyperuricemia (HU) is a well recognized risk factor for cardiovascular diseases. The independence of this association from other confounding factors has remained controversial. But the possible contributory effect of HU to carotid intima-media thickness (C-IMT) produced by hypertension (HT) has not been clarified yet. The study was designed to assess the C-IMT in patients with hypertension (HT) with or without HU. METHODS: The study participants consisted of 30 patients (men 60%, mean age+/-S.D.: 49+/-11 years) with HT without HU, and 25 patients with HT and HU (men 52%, mean age+/-S.D.: 52+/-12 years), and 25 age-matched healthy control subjects (men 56%, mean age+/-S.D.: 50+/-13 years). All study groups were examined by B-mode ultrasound to measure the C-IMT at the far wall of the common carotid artery. RESULTS: C-IMT were significantly higher in the patient groups (HT without HU and HT with HU) compared to the control cases (0.70+/-0.14, 0.83+/ 0.16 versus 0.57+/-0.16, mm, respectively, p<0.001). In the patients groups, patients with HU had significantly higher carotid IMT compared to the patients without HU. In stepwise linear regression analysis, we found that serum uric acid (SUA) levels independently but modestly associated with C-IMT (beta=0.42, p=0.002). CONCLUSION: We have shown that higher SUA levels are associated with atherogenesis independent from hypertension. Prospective studies will be necessary to confirm and extend these findings including early screening for hyperuricemia and lowering of SUA level looking at potential benefits in slowing progression of C-IMT in hypertensive patients. PMID- 17416372 TI - Levels of extra domain A containing fibronectin in human atherosclerotic plaques are associated with a stable plaque phenotype. AB - BACKGROUND: Extra domain A (EDA), splice-variant of fibronectin, is a Toll-like receptor 4 (Tlr4) ligand. Recently, EDA has been demonstrated to enhance atherogenesis in mice but human data on the role of EDA in atherosclerotic disease are lacking. We hypothesized that EDA is associated with unstable plaque phenotypes and that plasma EDA could serve as biomarker for atherosclerosis. METHODS: EDA levels were assessed in carotid endarterectomy specimen (206 patients) and related with plaque phenotype. In a second patient cohort, systemic EDA levels in atherosclerotic patients (73 patients) were compared to risk-factor matched controls (68 patients). RESULTS: EDA plaque levels were associated with characteristics of stable plaques; more smooth muscle cells (P=0.003), more collagen (P=0.071) and less fat (P=0.023). Concomitantly, asymptomatic patients showed higher EDA values in the plaque compared to symptomatic patients (P=0.004). EDA plasma levels did not differ between atherosclerotic patients versus controls (P=0.134). CONCLUSION: EDA plaque levels are higher in asymptomatic patients and are associated with a stable plaque phenotype. EDA is not a plasma marker for atherosclerotic disease. These results suggest that local presence of endogenous Tlr4 ligand EDA is not associated with in an unstable plaque phenotype in humans. PMID- 17416373 TI - Direct and indirect high-performance liquid chromatography enantioseparation of trans-4-hydroxy-2-nonenoic acid. AB - trans-4-Hydroxy-2-nonenoic acid (HNEA) is a marker of lipid peroxidation resulting from the metabolism of trans-4-hydroxy-2-nonenal (HNE). Direct and indirect RP-HPLC methods for the separation of HNEA enantiomers were developed and compared. The indirect method involved pre-column derivatization with a chiral amino agent, (1S,2S)-(+)-2-amino-1-(4-nitrophenyl)-1,3-propanediol, and subsequent separation of diastereomers on a Spherisorb ODS2 column. The direct separation of HNEA enantiomers was performed using the chiral stationary phase, Chiralpak AD-RH. Validation parameters including limit of quantification, linear range, accuracy and precision were determined. The indirect separation method was successfully applied for the determination of enantiomeric ratio of HNEA in rat brain mitochondrial lysate, and showed that HNEA was formed (R) enantioselectively from HNE. PMID- 17416374 TI - Normal phase high performance liquid chromatography for fractionation of organic acid mixtures extracted from crude oils. AB - Crude oil contains such an extensive range of compounds that a complete analysis is impossible. Fractionation by chemical properties is often used to simplify analytical handling. This work presents a high performance liquid chromatography (HPLC) method using normal phase chromatography on a cyano-bonded phase column to separate acid extracts from crude oils into four fractions; non-polar compounds, saturated carboxylic acids, phenols and polyfunctional acids. The method has been developed both in analytical scale for characterisation of acid extracts, and in preparative scale to provide sufficient sample amounts for further analysis by complementary methods. PMID- 17416375 TI - Towards unsupervised analysis of second-order chromatographic data: automated selection of number of components in multivariate curve-resolution methods. AB - A method to apply multivariate curve-resolution unattendedly is presented. The algorithm is suitable to perform deconvolution of two-way data (e.g. retrieving the individual elution profiles and spectra of co-eluting compounds from signals obtained from a chromatograph equipped with multiple-channel detection: LC-DAD or GC-MS). The method is especially adequate to achieve the advantages of deconvolution approaches when huge amounts of data are present and manual application of multivariate techniques is too time-consuming. The philosophy of the algorithm is to mimic the reactions of an expert user when applying the orthogonal projection approach--multivariate curve-resolution techniques. Basically, the method establishes a way to check the number of significant components in the data matrix. The performance of the method was superior to the Malinowski F-test. The algorithm was tested with HPLC-DAD signals. PMID- 17416376 TI - Trace analysis of total fluorine in human blood using combustion ion chromatography for fluorine: a mass balance approach for the determination of known and unknown organofluorine compounds. AB - The number of perfluorochemicals (PFCs) that have been found in biological and environmental matrices is increasing as analytical standards and methods evolve. Perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) constitute only a fraction of the total suite of PFCs found in environmental and biological matrices. A robust method and approach is needed to evaluate the mass of fluorinated compounds in biological matrices. In this study, we developed a method to measure total fluorine (TF) and organic fluorine (TOF) in human blood matrices using combustion ion chromatography (CIC). Blood matrices (whole blood, serum, and plasma) were analyzed in bulk to determine TF. An aliquot of the blood was also extracted with organic solvents such as methyl-tert-butyl ether (MTBE) and hexane, and organic and aqueous extracts were separated, to fractionate organofluorines from inorganic fluorine. The organic layer was analyzed for TF by CIC, and for known PFCs by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). PFCs measured by HPLC-MS/MS accounted for >80% of the TF in the organic fraction. The aqueous fraction contained inorganic fluorine and other non-extractable organofluorines. However, in the bulk sample, fluoride and non-extractable organofluorines accounted for >70% of the TF in blood samples from the general population. In occupationally exposed individuals, known organofluorines accounted for a major proportion of the TF. These results suggest the existence of yet uncharacterized fluorine fraction in human blood. Further studies are needed to characterize the aqueous fraction that contains inorganic fluorine and non-extractable forms of fluorine. PMID- 17416377 TI - Statistical designs and response surface techniques for the optimization of chromatographic systems. AB - This paper describes fundamentals and applications of multivariate statistical techniques for the optimization of chromatographic systems. The surface response methodologies: central composite design, Doehlert matrix and Box-Behnken design are discussed and applications of these techniques for optimization of sample preparation steps (extractions) and determination of experimental conditions for chromatographic separations are presented. The use of mixture design for optimization of mobile phases is also related. An optimization example involving a real separation process is exhaustively described. A discussion about model validation is presented. Some applications of other multivariate techniques for optimization of chromatographic methods are also summarized. PMID- 17416378 TI - Immunoaffinity centrifugal precipitation chromatography. AB - Purification of proteins based on immunoaffinity has been performed using a solid support coated with antibody against the target proteins. The method requires immobilizing the antibody onto the solid support using protein A or G, and has a risk of adsorptive loss of target proteins onto the solid support. Centrifugal precipitation chromatography has been successfully used to purify enzymes, such as ketosteroid isomerase and hyaluronidase without the use of solid support. The purpose of this study is to demonstrate that immunoaffinity centrifugal precipitation chromatography is capable of isolating an antigen by exploiting antigen-antibody binding. The separation was initiated by filling both channels with 40% saturated ammonium sulfate (AS) of pH 4-4.5 followed by loading 20 microl of human plasma (National Institutes of Health blood bank) mixed with 2 mg of rabbit anti-HSA (human serum protein) antibody (Sigma). Then, the sample channel was eluted with water at 0.03 ml/min and AS channel with 40% AS solution of pH 4-4.5 at 1 ml/min until all non-binding components were eluted. Then, the releasing reagent (50% AS solution containing 0.5 M glycine and 10% ammonium hydroxide at pH 10) was introduced through the AS channel to release the target protein (HSA). The retained antibody was recovered by eluting the sample channel with water at 1 ml/min. A hollow fiber membrane device at the outlet (MicroKros, Spectrum, New Brunswick, NJ, USA) was provided on-line dialysis of the eluent before fractions were collected, so that the fractions could be analyzed by SDS PAGE (sodium dodecyl sulfate - polyacrylamide gel electrophoresis) without further dialysis. The current method does not require immobilizing the antibody onto a matrix, which is used by the conventional immunoaffinity chromatography. This method ensures full recovery of the antigen and antibody, and it may be applied to purification of other proteins. PMID- 17416379 TI - Development and validation of a high-performance liquid chromatographic method using fluorescence detection for the determination of vardenafil in small volumes of rat plasma and bile. AB - A new, simple and sensitive high-performance liquid chromatography (HPLC) method with fluorescence detection was developed and validated for the determination of vardenafil in small volumes of rat plasma and bile. The absorbance and fluorescence characteristics of vardenafil were studied and factors that affect the HPLC resolution and fluorescence intensity were examined and optimized. Vardenafil and the internal standard cisapride were extracted using acetonitrile. The separation was achieved on a C18 column at 35 degrees C using acetonitrile-50 mM ammonium acetate aqueous solution (pH 6.8) (40:60) as mobile phase. At a flow rate of 1 ml/min, the total run time was 18 min. Fluorescence was measured with excitation and emission set at 280 and 470 nm, respectively. The calibration curves were linear from 10 to 1000 ng/ml and 0.2-100 microg/ml for plasma and bile samples, respectively. The intra- and inter-day imprecision did not exceed 10.8%, and the accuracy was within 9.6% deviation of the nominal concentration. The method was used successfully to investigate the disposition and biliary excretion of vardenafil in rats. PMID- 17416380 TI - Development and validation of a multi-residue method for pesticide determination in honey using on-column liquid-liquid extraction and liquid chromatography tandem mass spectrometry. AB - We report on the development and validation under ISO 17025 criteria of a multi residue confirmatory method to identify and quantify 17 widely chemically different pesticides (insecticides: Carbofuran, Methiocarb, Pirimicarb, Dimethoate, Fipronil, Imidacloprid; herbicides: Amidosulfuron, Rimsulfuron, Atrazine, Simazine, Chloroturon, Linuron, Isoxaflutole, Metosulam; fungicides: Diethofencarb) and 2 metabolites (Methiocarb sulfoxide and 2 Hydroxytertbutylazine) in honey. This method is based on an on-column liquid liquid extraction (OCLLE) using diatomaceous earth as inert solid support and liquid chromatography (LC) coupled to mass spectrometry (MS) operating in tandem mode (MS/MS). Method specificity is ensured by checking retention time and theoretical ratio between two transitions from a single precursor ion. Linearity is demonstrated all along the range of concentration that was investigated, from 0.1 to 20 ng g(-1) raw honey, with correlation coefficients ranging from 0.921 to 0.999, depending on chemicals. Recovery rates obtained on home-made quality control samples are between 71 and 90%, well above the range defined by the EC/657/2002 document, but in the range we had fixed to ensure proper quantification, as levels found in real samples could not be corrected for recovery rates. Reproducibility is found to be between 8 and 27%. Calculated CCalpha and CCbeta (0.0002-0.943 ng g(-1) for CCalpha, and 0.0002-1.232 ng g(-1) for CCbeta) show the good sensitivity attained by this multi-residue analytical method. The robustness of the method has been tested in analyzing more than 100 raw honey samples collected from different areas in Belgium, as well as some wax and bee samples, with a slightly adapted procedure. PMID- 17416381 TI - Microemulsion electrokinetic chromatography with on-line atmospheric pressure photoionization-mass spectrometric detection of medium polarity compounds. AB - In this paper, we present the determination of pharmaceuticals employing microemulsion electrokinetic chromatography (MEEKC) with atmospheric pressure photoionization-mass spectrometric (APPI-MS) detection. This recent hyphenated technique allows to overcome some disadvantages of MEEKC, namely its inherent incompatibility with MS detection. Important parameters like microemulsion (ME) composition, the composition of the sheath liquid and APPI-MS detection parameters have been investigated. Using the optimized set of parameters, the eight selected substances could be detected down to concentrations between 3 mg L(-1) (phenacetin) and 41 mg L(-1) (diltiazem). Switching to the MS2 mode, the use of specific transitions for the detection of each analyte provided improved detection limits in the range of 0.6 mg L(-1) (carbamazepine) to 6 mg L(-1) (metoprolol). Calibration curves were linear over one to two orders of magnitude with correlation coefficients better than 0.98. PMID- 17416382 TI - Effects of capillary coating and beta-cyclodextrin additive to the background electrolyte on separation of sulphonated azodyes by capillary zone electrophoresis. AB - Fourteen azodyes containing one to five acidic groups were separated by capillary zone electrophoresis (CZE). The effects of the pH and beta-cyclodextrin additive to the background electrolyte on the separation of sulphonated azodyes were investigated. The effects of the working conditions significantly differ in non coated fused silica capillaries and in capillaries coated with polyacrylamide. Splitting of the zones of metal-complex dyes was observed in polyacrylamide coated capillaries and the background electrolyte with 10 mmol/L beta cyclodextrin, due to the separation of stereoisomeric forms of the dyes, which were separated for the first time using CE. Relations between the structure of the sulphonated azodyes and the electrophoretic mobilities are discussed. Naphthalene mono- to tetrasulphonic acids were used as the standards for the calibration of migration scale of the analysed dyes. PMID- 17416383 TI - In situ functionalization of N-acryloxysuccinimide-based monolith for reversed phase electrochromatography. AB - Capillary electrochromatography (CEC) monolithic columns were prepared following a two-step synthetic pathway based on (i) UV-induced in situ radical polymerization of N-acryloxysuccinimide (NAS) and ethylene dimethacrylate (EDMA) and (ii) in situ functionalization of the NAS-containing monolithic matrix with various alkylamines. The first synthetic step was performed using toluene as a porogenic solvent. The successful grafting of the alkylamines onto the reactive matrix was confirmed on the basis of qualitative analysis of Raman spectra recorded before and after the chemical modification step. All the electrochromatographic results indicate a strong dependence of the retention, efficiency and selectivity of the monolithic columns on small variations of mobile phase composition and nature of the grafted aliphatic selector in agreement with the typical reversed-phase behaviour. Van Deemter plots for a series of alkylbenzene homologues injected on a column bearing hexyl-segments as side chains are also presented. PMID- 17416384 TI - Electric field induced instabilities in thin confined bilayers. AB - A long wave nonlinear theory and simulations on the electric field induced instability of a thin (<1000 nm thick) viscous bilayer resting on a solid substrate are presented. The instabilities in these systems are initiated by one of the two basic short time modes of deformation at the twin interfaces-in-phase bending or out of phase squeezing. Linear stability analysis (LSA) is carried out to identify the conditions for these modes. It is shown that these modes can be switched and the relative amplitudes of deformation at the interfaces can be profoundly altered by varying the thicknesses, viscosities, interfacial tensions and dielectric constants of the films. Nonlinear simulations are presented to support the results obtained from the LSA. In addition, simulations show a number of interesting interfacial morphologies including: (a) embedded upper layer in the array of lower layer columns, (b) columns of the upper layer grown towards the substrate and sheathed by the lower layer liquid, (c) lower layer columns sheathed by the upper layer liquid leading to concentric core-shell columns, (d) droplets of upper liquid on the largely undisturbed lower layer, (e) symmetry breaking traveling waves at the interfaces, and (f) evolution of two different wavelengths at the two interfaces of a bilayer. The effects of viscous and the capillary resistances on the evolution of instability and morphology are also discussed. PMID- 17416385 TI - Modification of silica gel by attachment of 2-mercaptobenzimidazole for use in removing Hg(II) from aqueous media: a thermodynamic approach. AB - The compound 2-mercaptobenzimidazole (MBI) was attached onto a silica gel surface by homogeneous and heterogeneous routes. Both silica modification methodologies resulted in similar products, named SiM(hom) and SiM(het), respectively. These materials were characterized by surface area, infrared, thermogravimetry, and 13C and 29Si NMR spectroscopy. The ability of these materials to remove divalent mercury from aqueous solution was followed by a series of adsorption isotherms adjusted to a modified Langmuir equation. The maximum number of moles adsorbed was determined to be 1.35+/-0.3 and 1.42+/-0.17 mmolg-1 for SiMhet and SiMhomt, respectively. These interactions were calorimetrically followed and the thermodynamic data showed the following exothermic enthalpic values: -23.04+/-1.4 and -20.08+/-1.2 kJmol-1 for SiMhom and SiMhet, respectively. All liquid/solid interface adsorptions were spontaneous in nature and enthalpically driven. PMID- 17416386 TI - Enhancement of photocatalytic activity of P25 TiO2 by vanadium-ion implantation under visible light irradiation. AB - An ion-implantation method was used to prepare V-ion-implanted P25 TiO2 photocatalysts. Their photocatalytic activity for the degradation of formic acid under visible light irradiation (lambda>450 nm) was investigated. Upon implantation of V ions into the lattice of P25 TiO2, the photoactivity was remarkably enhanced. HRTEM images showed that the implanted V ions existed in the form of VO2(T) in the lattice of P25 TiO2. The intensity of photoluminescence (PL) spectra of V-ion-implanted P25 TiO2 decreased with the increase of the amount of implanted V ions, indicating the decrease of electron-hole pair recombination. It was also observed that the lower the PL intensity of V-ion implanted P25 TiO2, the higher the photoactivity. PMID- 17416387 TI - Microstructure and structural transition in coconut oil microemulsion using semidifferential electroanalysis. AB - The microemulsification in coconut oil/octadecyltrimethylammonium bromide/iso pentanol/water system is investigated. The effect of iso-pentanol concentrations on the size of single-phase microemulsion region is discussed. It is found that the maximum microemulsion domain is obtained when cosurfactant (iso-pentanol)-to surfactant (octadecyltrimethylammonium bromide) mass ratio is 1.5. The diffusion coefficients of ferrocene (electroactive probe) in microemulsion microenvironment are measured by semidifferential electroanalysis. The microstructure and structural transition from water-in-oil to oil-in-water microemulsions through a bicontinuous structure is examined. The results are found to be in agreement with that of conductivity measurements. PMID- 17416388 TI - Effects of 3-amino-1,2,4-triazole on the inhibition of copper corrosion in acidic chloride solutions. AB - Effects of 3-amino-1,2,4-triazole (ATA) on the inhibition of copper corrosion in 0.5 M HCl solutions have been studied using gravimetric, electrochemical, and Raman spectroscopy investigations. Weight-loss measurements after varied immersion periods revealed that the dissolution rate of copper decreased to a minimum, while the inhibition efficiency (zeta%) and consequently the degree of surface coverage (theta) increased with the presence of ATA and the increase of its concentration. Potentiodynamic polarization, chronoamperometric, and electrochemical impedance spectroscopy (EIS) measurements after 0, 24, and 48 h immersion of the copper electrode in the test solutions showed that the presence of ATA molecules significantly decreased cathodic, anodic, and corrosion (jcorr) currents and corrosion rates (Rcorr) and greatly increased polarization resistance (Rp), zeta%, and theta; this effect was increased on increasing the ATA content in the solution. Raman spectroscopy confirmed that ATA molecules strongly adsorbed onto the copper surface, blocking its active sites and preventing it from being corroded easily. PMID- 17416389 TI - Effects of dynamics on ecological networks. AB - Ecological food webs define the feeding patterns of interacting species. The architecture of such networks may be affected by dynamical processes operating within them, ultimately influencing the capacity of the networks to persist. As yet relatively little is known about these effects. We compared the architecture of ecological networks with a fixed number of species, constructed in four contrasting ways: (I) topological networks, which required only that species had prey to eat; (II) persistent networks, in which species had also to persist under a simple model of population dynamics; (III) assembled networks, built up by sequential addition of species with dynamical persistence at each step in the sequence; (IV) evolved networks where, in addition to dynamical persistence, body size of species was determined by a simple mutation-selection process. Dynamics had fundamental effects on architecture, the networks of classes II, III and IV being restricted to a small number of trophic levels, in contrast to the non dynamic, topological class I networks. Class III assembled networks tended to have fewer trophic levels and a more pyramidal biomass distribution than networks of classes II and IV. In evolved class IV networks, the smallest consumers converged to similar body sizes, whereas larger consumers evolved more slowly and did not show such convergence. The results indicate that dynamics affect the architecture of food webs, and that assumptions about simultaneous arrival, sequential arrival and evolution lead to different outcomes. Sequential assembly was shown to have a special property of finding rare sets of persistent species in a small number of steps, suggesting that the rarity of stable communities is not a serious problem in the development of complex communities. PMID- 17416390 TI - An immersed boundary framework for modelling the growth of individual cells: an application to the early tumour development. AB - A biomechanical approach in modelling the growth and division of a single fully deformable cell by using an immersed boundary method with distributed sources is presented, and its application to model the early tumour development is discussed. This mathematical technique couples a continuous description of a viscous incompressible cytoplasm with the dynamics of separate elastic cells, containing their own point nuclei, elastic plasma membranes with membrane receptors, and individually regulated cell processes. This model enables one to focus on the biomechanical properties of individual cells and on communication between cells and their microenvironment, simultaneously allowing for the formation of clusters or sheets of cells that act together as one complex tissue. Several examples of early tumours growing in various geometrical configurations and with distinct conditions of their initiation and progression are also presented to show the strength of our approach in modelling different topologies of the growing tissues in distinct biochemical conditions of the surrounding media. PMID- 17416391 TI - Prisoner's dilemma posed by fitness-associated recombination strategies. AB - Genetic recombination is a central and repeated topic of study in the evolution of life. However, along with the influence of recombination on evolution, we understand surprisingly little of how selection shapes the nature of recombination. One explanation for recombination is that it allows organisms to escape from perilous situations where they experience very low fitness. As a corollary, it has been suggested that selection should favor recombination at low fitness and not at high fitness (fitness-associated recombination, FAR), and theory suggests that such strategies can indeed be selected. Here we develop models to further investigate the evolution of FAR. Consistent with previous works, we find that FAR can invade and dominate over a strategy of uniform recombination that is independent of fitness. However, our simulation results suggest that extreme FAR strategies, known as group-elitism, are not necessarily superior to other FAR strategies. Moreover, we argue that FAR domination will often occur with a net loss of mean population fitness. Interestingly, this suggests that the strategy of not recombining at high fitness will sometimes be analogous to a defector strategy from the famous "prisoner's dilemma" game: a selfish strategy that is selected but leads to a loss of mean fitness for all players. PMID- 17416392 TI - Determination of the optimal therapeutic protocols in cancer immunotherapy. AB - Cancer immunotherapy aims at eliciting an immune system response against the tumor. However, it is often characterized by toxic side-effects. Limiting the tumor growth and, concurrently, avoiding the toxicity of a drug, is the problem of protocol design. We formulate this question as an optimization problem and derive an algorithm for its solution. Unlike the standard optimal control approach, the algorithm simulates impulse-like drug administrations. It relies on an exact computation of the gradient of the cost function with respect to any protocol by means of the variational equations, that can be solved in parallel with the system. In comparison with previous versions of this method [F. Castiglione, B. Piccoli, Optimal control in a model of dendritic cell transfection cancer immunotherapy, Bull. Math. Biol. 68 (2006) 255-274; B. Piccoli, F. Castiglione, Optimal vaccine scheduling in cancer immunotherapy, Physica A. 370 (2) (2007) 672-680], we optimize both the timing and the dosage of each administration and introduce a penalty term to avoid clustering of subsequent injections, a requirement consistent with the clinical practice. In addition, we implement the optimization scheme to simulate the case of multi therapies. The procedure works for any ODE system describing the pharmacokinetics and pharmacodynamics of an arbitrary number of therapeutic agents. In this work, it was tested for a well known model of the tumor-immune system interaction [D. Kirschner, J.C. Panetta, Modeling immunotherapy of tumor-immune interaction, J. Math. Biol. 37 (1998) 235-252]. Exploring three immunotherapeutic scenarios (CTL therapy, IL-2 therapy and combined therapy), we display the stability and efficacy of the optimization method, obtaining protocols that are successful compromises between various clinical requirements. PMID- 17416393 TI - Prophylaxis or treatment? Optimal use of an antiviral stockpile during an influenza pandemic. AB - We introduce a novel mathematical model that effectively incorporates contact tracing in a realistic distribution mechanism for antiviral drugs in an influenza pandemic scenario. A strategy focused on targeted provision of post-exposure prophylaxis, rather than treatment, will provide the greatest chance of minimising the impact of an influenza pandemic. Targeted post-exposure prophylaxis delays the onset of the pandemic and for a wide range of parameter values, a delay of the order of 6-18 months may be achievable. This may provide enough time to develop and distribute a vaccine. In contrast, a treatment based strategy typically does not delay the onset of a pandemic by an appreciable amount and, in general, is not capable of significantly reducing the attack rate from baseline. PMID- 17416394 TI - Rubbish index and diarrhoea in Salvador, Brazil. AB - Urban solid waste (USW) is a public health problem worldwide that has not yet been completely evaluated in relation to childhood health. The main purpose of this study was to investigate associations between diarrhoea and exposure to USW among preschool children. A longitudinal study was carried out with 922 under threes living in Salvador, Bahia, between 2000 and 2001. Both community and individual/home levels (strata) were evaluated by questionnaire and direct observation. The multilevel model, using Poisson regression with overdispersion, evaluated their contribution to the occurrence of diarrhoea. The children presented a mean longitudinal prevalence of 2.7% of days with diarrhoea during a mean period of 23 follow-up weeks. The main factors associated with diarrhoea were rubbish-in-street index [prevalence ratio (PR)=1.30; 95% CI 1.02-1.66] and intrahome rubbish packaging (PR=3.59; 95% CI 2.01-6.42). Other important variables were water-in-street index, child age, hygiene and cleanliness near the house, number of people per house, and drainage problems. The variables measured at the community level explained 4.77% of the variability of the children's number of days with diarrhoea. PMID- 17416395 TI - Dynamics of infection with multiple transmission mechanisms in unmanaged/managed animal populations. AB - Deterministic and stochastic models motivated by Salmonella transmission in unmanaged/managed populations are studied. The SIRS models incorporate three routes of transmission (direct, vertical and indirect via free-living infectious units in the environment). With deterministic models we are able to understand the effects of different routes of transmission and other epidemiological factors on infection dynamics. In particular, vertical transmission has little influence on this dynamics, whereas the higher the indirect (direct) transmission rate the greater the tendency to persistent oscillation (stable endemic states). We show that the sustained cycles are also prone to demographic effect, i.e., persistent oscillation becomes impossible in the managed case (in the sense of balanced recruitment and death rates) by comparing with results in unmanaged populations (exponential population dynamics). Further, approximations of quasi-stationary distributions are derived for stochastic versions of the proposed models based on a diffusion approximation to the infection process. The effect of transmission parameters on the ratio of mean to standard deviation of the approximating distribution, used to judge the validity of the approximations and the expected time until fade out of infection, is further discussed. We conclude that strengthening any route of transmission may or may not reduce the expected time to fade out of infection, depending on the population dynamics. PMID- 17416397 TI - Bilateral long-range interaction between right and left visual hemifield. AB - Long-range interaction has been reported to be limited in space within a few degrees. Here, we present a new type of interaction by means of a bilateral configuration using two Gabor signals (GSs). Two horizontally oriented GSs, the first defined as a cue and the second as a probe, appeared in the right and left peripheral visual fields in mirror symmetrical regions. The detection threshold of the GS probe was found to decrease significantly up to cue-probe separations of 10 degrees tested. Since the interaction was sensitive to symmetrical locus, as well as specific to the direction of the horizontal axis, the results suggest novel long-range interaction extending toward the periphery with a mirror symmetrical configuration. This may be acquired by neuronal communication, which directly connects bilateral receptive fields in the right and left visual cortices. PMID- 17416396 TI - Normal ocular development in young rhesus monkeys (Macaca mulatta). AB - PURPOSE: The purpose of this study was to characterize normal ocular development in infant monkeys and to establish both qualitative and quantitative relationships between human and monkey refractive development. METHODS: The subjects were 214 normal rhesus monkeys. Cross-sectional data were obtained from 204 monkeys at about 3 weeks of age and longitudinal data were obtained from 10 representative animals beginning at about 3 weeks of age for a period of up to 5 years. Ocular development was characterized via refractive status, corneal power, crystalline lens parameters, and the eye's axial dimensions, which were determined by retinoscopy, keratometry, phakometry and A-scan ultrasonography, respectively. RESULTS: From birth to about 5 years of age, the growth curves for refractive error and most ocular components (excluding lens thickness and equivalent lens index) followed exponential trajectories and were highly coordinated between the two eyes. However, overall ocular growth was not a simple process of increasing the scale of each ocular component in a proportional manner. Instead the rates and relative amounts of change varied within and between ocular structures. CONCLUSION: The configuration and contribution of the major ocular components in infant and adolescent monkey eyes are qualitatively and quantitatively very comparable to those in human eyes and their development proceeds in a similar manner in both species. As a consequence, in both species the adolescent eye is not simply a scaled version of the infant eye. PMID- 17416398 TI - Induced internal noise in perceptual artificial scotomas created by surrounding dynamic noise. AB - Research has shown that exposure to a homogeneous gray patch surrounded by a dynamic noise background causes filling-in of the artificial scotoma by the twinkling noise from the surround. When the background is switched off, observers report perception of a prolonged patch of twinkling noise in the unstimulated area. We studied the effects of exposure to a centrally presented artificial scotoma and the twinkling aftereffect on the threshold for detecting a foveal Gabor patch embedded in external scotoma noise. The detection thresholds were mainly elevated in the absence of scotoma noise and less affected at higher levels of scotoma noise. The analysis of the experimental data using the equivalent input noise approach revealed that the reduced contrast sensitivity is due to induced internal noise whose variance is proportional to the strength of the surrounding noise. We did not find significant effects on the internal noise in a control experiment using flickering Gaussian noise samples of 1.6 Hz which did not cause filling-in and dynamic afterimage. These findings suggest that the perceptual phenomena caused by artificial scotomas may reflect increased variability of neural activity due to long-range interactions between the surrounding noise and unstimulated region of the artificial scotoma. PMID- 17416399 TI - Influence of biofilms on the movement of colloids in porous media. Implications for colloid facilitated transport in subsurface environments. AB - Colloid transport through porous media can be influenced by the presence of biofilms. Sterile and non-sterile sand columns were investigated using Laponite RD as model colloid and a highly mucoid strain of Pseudomonas aeruginosa as model biofilm former. Laponite RD was marked specifically by fluorescent complexes with rhodamine 6G. Breakthrough curves (BTCs) were used as parameters for determination of colloid transport characteristics. In the sterile columns, the colloid was mobile (collision efficiencies from 0.05 to 0.08) both after the presence of Na(+) and Ca(2+) ions followed by deionised water influent. In the biofilm-grown column, the same treatment did not result in colloid retention in the case of Na(+) exposure, but in altered or enhanced colloid transport. In the case of Ca(2+) ions exposure, colloid retention increased with biofilm age. After 3 weeks, almost complete retention was observed. Similar observations were made in columns packed with material from slow sand filtration units. These data reveal the complex interactions between biofilms, cations and colloid transport. Changes in the electrolyte composition of water percolating the subsurface can frequently occur and will result in different colloid transport characteristics with regard to the dominating species of ions and the relative abundance of microbial biofilms. This has to be considered when modelling colloid transport through the subsurface. PMID- 17416400 TI - Persulfate-induced photochemical decomposition of a fluorotelomer unsaturated carboxylic acid in water. AB - The persulfate (S(2)O(8)(2-))-induced photochemical decomposition of C(3)F(7)CF=CHCOOH in water was investigated to develop a method to neutralize stationary sources of fluorotelomer unsaturated carboxylic acids (FTUCAs), which have recently been detected in the environment, and are considered to be more toxic than the environmentally persistent perfluorocarboxylic acids (PFCAs). Photolysis of S(2)O(8)(2-) produced highly oxidative sulfate radical anions (SO(4)(-)), which efficiently decomposed C(3)F(7)CF=CHCOOH to F(-) and CO(2) via C(3)F(7)COOH. With an initial S(2)O(8)(2-) concentration of 12.5mM and irradiation from a 200-W xenon-mercury lamp, C(3)F(7)CF=CHCOOH at a concentration of 680 microM was completely decomposed within 5 min. When 8.00 mM S(2)O(8)(2-) was used, the initial rate of C(3)F(7)CF=CHCOOH decomposition induced by 254-nm light irradiation was 45 times as high as that with photolysis alone. The apparent quantum yield for the C(3)F(7)CF=CHCOOH decomposition with 6.25 mM S(2)O(8)(2-) and 254-nm light was 2.4, indicating that virtually all SO(4)(-) anions produced by the photolysis of S(2)O(8)(2-) contribute to the decomposition of C(3)F(7)CF=CHCOOH. PMID- 17416401 TI - Comparison of natural estrogen removal efficiency in the conventional activated sludge process and the oxidation ditch process. AB - The presence of natural estrogens, 17beta-estradiol (E2), estrone (E1) and estriol (E3), as well as estrogenic activity in wastewater influents and secondary effluents were investigated in 20 full-scale wastewater treatment plants in Japan. In all of the influent samples, natural estrogens were detected at concentrations above the minimum limits of detection (0.5ng/L). The concentrations of natural estrogens detected in the effluent of oxidation ditch plants were generally lower than previously reported values. On the other hand, in the conventional activated sludge plants, increments of E1 during biological treatment were frequently observed although E2 and E3 were removed effectively in the process. The removal rates of natural estrogens or estrogenic activity show no observed statistical relationship with the solids retention time (SRT) and the hydraulic retention time (HRT). However, the plants with high SRT or HRT generally showed high and stable removal of both natural estrogens and estrogenic activity. PMID- 17416402 TI - Studies on the restoration succession of PFU microbial communities in a pilot scale microcosm. AB - In order to imitate the restoration succession process of natural water ecosystem, a laboratory microcosm system of constant-flow-restoration was designed and established. A eutrophycation lake, Lake Donghu, was selected as the subject investigated. Six sampling stations were set on the lake, among which the water of station IV was natural clean water, and others were polluted with different degrees. Polyurethane foam unit microbial communities, which had colonized in the stations for a month, were collected from these stations and placed in their respective microcosms, using clean water of station IV to gradually replace the water of these microcosms. In this process, the healthy community in clean water continuously replaced the damaged communities in polluted water, the restoration succession of the damaged communities was characterized by weekly determination of several functional and structural community parameters, including species number (S), diversity index (DI), community pollution value (CPV), heterotrophy index (HI), and similarity coefficient. Cluster analysis based on similarity coefficient was used to compare the succession discrepancies of these microbial communities from different stations. The ecological succession of microbial communities during restoration was investigated by the variable patterns of these parameters, and based on which, the restoration standards of these polluted stations were suggested in an ecological sense. That was, while being restored, the water of station 0 (supereutrophycation) should be substituted with natural clean water by 95%; station I (eutrophycation), more than 90%; station II (eutrophycation), more than 85%; station III (eutrophycation), about 85%; station V (mesoeutrophycation), less than 50%. The effects of the structural and functional parameters in monitoring and assessing ecological restoration are analyzed and compared. PMID- 17416403 TI - Toxicity of ferric chloride sludge to aquatic organisms. AB - Iron-rich sludge from a drinking water treatment plant (DWTP) was investigated regarding its toxicity to aquatic organisms and physical and chemical composition. In addition, the water quality of the receiving stream near the DWTP was evaluated. Experiments were carried out in August 1998, February 1999 and May 1999. Acute toxicity tests were carried out on a cladoceran (Daphnia similis), a midge (Chironomus xanthus) and a fish (Hyphessobrycon eques). Chronic tests were conducted only on D. similis. Acute sludge toxicity was not detected using any of the aquatic organisms, but chronic effects were observed upon the fecundity of D. similis. Although there were relatively few sample dates, the results suggested that the DWTP sludge had a negative effect on the receiving body as here was increased suspended matter, turbidity, conductivity, chemical oxygen demand (COD) and hardness in the water downstream of the DWTP effluent discharge. The ferric chloride sludge also exhibited high heavy metal concentrations revealing a further potential for pollution and harmful chronic effects on the aquatic biota when the sludge is disposed of without previous treatment. PMID- 17416404 TI - Elimination of racemic and enantioenriched metalaxyl based fungicides under tropical conditions in the field. AB - The elimination has been studied of racemic and enantioenriched metalaxyl applied as an emulsifiable concentrate and wettable powder combined with copper in a Cameroonian field site. The kinetics of the degradation/dissipation of metalaxyl and its acid metabolite were investigated using reversed phase HPLC-MS/MS, while the enantiomeric ratios were measured by HPLC-MS/MS using a Chiralcel OD-H HPLC column. Some soil enzymes activities were determined concurrently for 120d. The elimination of racemic metalaxyl was shown to be enantioselective, with the R enantiomer being degraded more slowly than the S-enantiomer. Dissipation followed approximate square root first-order kinetics (R>0.98) without lag phases. The enantiomers of metalaxyl have different elimination rates, with half-lives ranging from only 0.8 to 1.5 days. After application to soil, the elimination of metalaxyl in the copper containing formulation was slower. The activities of acid phosphatase, alkaline phosphatase, and alkaline glucosidase were monitored throughout the experiments. No significant influence of metalaxyl and copper could be observed on these parameters. The significantly shorter half-life values of all forms of metalaxyl under field conditions, compared to the previously reported laboratory derived ones, may have implications for the plant disease control with these fungicides in tropical rainforest areas. PMID- 17416405 TI - Enhanced phytoremediation of arsenic contaminated land. AB - In an attempt to clean up arsenic (As) contaminated soil, the effects of phosphorus (P) fertilizer and rhizosphere microbes on arsenic accumulation by the silverback fern, Pityrogramma calomelanos, were investigated in both greenhouse and field experiments. Field experiments were conducted in Ron Phibun District, an As-contaminated area in Thailand. Soil (136-269 microg As g(-1)) was collected there and used in the greenhouse experiment. Rhizosphere microbes (bacteria and fungi) were isolated from roots of P. calomelanos growing in Ron Phibun District. The results showed that P-fertilizer significantly increased plant biomass and As accumulation of the experimental P. calomelanos. Rhizobacteria increased significantly the biomass and As content of the test plants. Thus, P-fertilizer and rhizosphere bacteria enhanced As-phytoextraction. In contrast, rhizofungi reduced significantly As concentration in plants but increased plant biomass. Therefore, rhizosphere fungi exerted their effects on phytostabilization. PMID- 17416406 TI - CD57 expression correlates with alveolitis severity in subjects with beryllium induced disease. AB - BACKGROUND: Despite numerous studies in human beings linking surface phenotype of blood T cells with their functional characteristics, little is known about this relationship on antigen-specific CD4(+) T cells residing in a target organ. OBJECTIVE: The aims of this study were to determine the relationship between CD57 expression, a marker of T-cell senescence, and severity of chronic beryllium disease (CBD) and to determine the phenotypic and functional characteristics that differentiate beryllium-specific CD4(+) T cells in lung and blood. METHODS: CD57 expression on beryllium-responsive IFN-gamma-expressing and IL-2-expressing CD4(+) T cells in blood and lung of 17 beryllium-sensitized and 20 CBD subjects was determined. RESULTS: CD57 expression was significantly higher on bronchoalveolar lavage (BAL) than blood CD4(+) T cells in both beryllium sensitized and CBD subjects. Expression of CD57 on BAL CD4(+) T cells was directly correlated with the lymphocytic alveolitis. In blood and BAL, higher CD57 expression was seen on more differentiated CD4(+) memory T-cell subsets. Although CD57 expression on blood and BAL cells was associated with a reduced proliferative potential, examination of beryllium-specific CD4(+) T cells in blood and lung revealed no difference in CD57 expression on cells that produced IFN-gamma only versus IFN-gamma and IL-2. CONCLUSION: These data suggest that CD57 expression on CD4(+) T cells is an important phenotypic marker to assess lung inflammation and the functional competence of the CD4(+) T-cell compartment in CBD. CLINICAL IMPLICATIONS: These findings suggest that CD57 is a marker of lung inflammation and potentially, disease severity. PMID- 17416407 TI - Achieving and maintaining asthma control in an urban pediatric disease management program: the Breathmobile Program. AB - BACKGROUND: National guidelines suggest that, with appropriate care, most patients can control their asthma. The probabilities of children achieving and maintaining control with ongoing care are unknown. OBJECTIVE: We sought to evaluate the degree to which children in a lower socioeconomic urban setting achieve and maintain control of asthma with regular participation in a disease management program that provides guideline-based care. METHODS: Interdisciplinary teams of asthma specialists use mobile clinics to offer ongoing care at schools and county clinics. A guideline-derived construct of asthma control is recorded at each visit. RESULTS: Two thousand one hundred eighty-five enrollees were eligible to evaluate the time to first achieve control, and 1591 patients were eligible to evaluate subsequent control maintenance. Depending on severity, 70% to 87% of patients with persistent asthma achieved control by visit 3, and 89% to 98% achieved control by visit 6. Subsequent control maintenance was highly variable. Thirty-nine percent of patients displayed well-controlled asthma (control at >90% of subsequent visits), whereas 13% displayed difficult-to control asthma (<50% of subsequent visits). Patients from each baseline severity category were found in each group. Maintenance of control was influenced by physician-estimated compliance with the treatment plan, baseline severity, and the interval between clinic visits. CONCLUSIONS: Many children can achieve asthma control with regular visit intervals and guideline-based care; however, long-term control can be highly variable among patients in all severity categories. CLINICAL IMPLICATIONS: These findings highlight the need and feasibility for systematically tracking each patient's clinical response to individualize therapy and guide the use of population management strategies. PMID- 17416408 TI - The relationship between seroatopy and symptoms of either allergic rhinitis or asthma. AB - BACKGROUND: Epidemiologic data on allergic rhinitis and asthma are frequently based on self-reported symptoms. OBJECTIVE: This cross-sectional study examined the relationship between self-reported symptoms and histories of allergic rhinitis or asthma and a marker of allergic sensitization, allergen-specific IgE. METHODS: We surveyed 702 pregnant women in Michigan. Blood samples were analyzed for specific IgE to 9 allergens: dust mites (Dermatophagoides farinae and Dermatophagoides pteronyssinus), cat, dog, cockroach, ragweed, timothy grass, egg, and Alternaria alternata. Seratopy was defined as a specific IgE greater than or equal to 0.35 kU/L to any allergen. RESULTS: Seroatopy was found in 66.7% of those with hay fever symptoms, 68.3% with a physician's diagnosis of asthma, and 72.1% of those with both conditions. These results differed significantly from asymptomatic subjects, where 49.8% of patients without hay fever and 50.4% without asthma were seroatopic. Race and education did not modify the relationships. Symptoms related to specific exposures were modest predictors of positive specific IgE to related allergens (positive predictive values from 26.5% to 50.3%). CONCLUSION: Self-reported symptoms of allergic rhinitis or asthma were significantly associated with allergic sensitization, but the odds ratios were of relatively low magnitude for this historical information to be considered evidence of current allergic sensitization. A 66% to 68% probability existed that those with symptoms of allergic rhinitis or asthma would have a positive specific IgE test. CLINICAL IMPLICATIONS: Self-reported histories of hay fever or asthma alone are only modest predictors of allergic sensitization. When knowledge of allergic sensitization is important, information beyond self-reported symptoms is necessary. PMID- 17416409 TI - Severe asthma in adults: what are the important questions? AB - The term severe refractory asthma (SRA) in adults applies to patients who remain difficult to control despite extensive re-evaluation of diagnosis and management following an observational period of at least 6 months by a specialist. Factors that influence asthma control should be recognized and adequately addressed prior to confirming the diagnosis of SRA. This report presents statements according to the literature defining SRA in order address the important questions. Phenotyping SRA will improve our understanding of mechanisms, natural history, and prognosis. Female gender, obesity, and smoking are associated with SRA. Atopy is less frequent in SRA, but occupational sensitizers are common inducers of new-onset SRA. Viruses contribute to severe exacerbations and can persist in the airways for long periods. Inflammatory cells are in the airways of the majority of patients with SRA and persist despite steroid therapy. The T(H)2 immune process alone is inadequate to explain SRA. Reduced responsiveness to corticosteroids is common, and epithelial cell and smooth muscle abnormalities are found, contributing to airway narrowing. Large and small airway wall thickening is observed, but parenchymal abnormalities may influence airway limitation. Inhaled corticosteroids and bronchodilators are the mainstay of treatment, but patients with SRA remain uncontrolled, indicating a need for new therapies. PMID- 17416410 TI - Nuclear reprogramming in embryos generated by the transfer of yak (Bos grunniens) nuclei into bovine oocytes and comparison with bovine-bovine SCNT and bovine IVF embryos. AB - Although inter-species SCNT may be useful for increasing and preserving populations of endangered species, there are many reports that inter-species nuclear transfer embryos only develop to the blastocyst stage. In this study, yak bovine SCNT blastocysts were successfully implanted in the surrogate bovine uterus but failed to develop to term or aborted. To clarify the reasons, we examined yak-bovine SCNT blastocyst development, total cell number, inner cell mass (ICM) number, trophoblast (TE) cell number and relative gene expression in yak fibroblast cells and yak-bovine SCNT embryos at various stages. The potential for development of yak-bovine SCNT embryos to blastocysts was 30+/-5.7% (mean+/ S.E.M.); the total cell number was 85.3+/-16.3, fewer than in IVF bovine embryos (106.2+/-18.2) but within the reported range (60-300). The yak-bovine SCNT blastocysts had a lower ratio of TE cells to total cells (43.9+/-8.7%) than bovine IVF embryos (59.4+/-3.4%; P<0.05) or bovine-bovine SCNT (69.5+/-5.4%; P<0.05). Also, several yak-bovine SCNT embryos had abnormal initiation of expression of both Mash2 and IL6. However, expression of vimentin, collagen, Cx43 and PSMC3 were normal in yak fibroblast cells and yak-bovine SCNT embryos. In conclusion, we inferred that the normal allocation of ICM and TE cells in yak bovine SCNT embryos and embryo-specific gene reprogramming may be important for successful inter-species animal cloning. PMID- 17416411 TI - Neural evidence for the interplay between language, gesture, and action: a review. AB - Co-speech gestures embody a form of manual action that is tightly coupled to the language system. As such, the co-occurrence of speech and co-speech gestures is an excellent example of the interplay between language and action. There are, however, other ways in which language and action can be thought of as closely related. In this paper we will give an overview of studies in cognitive neuroscience that examine the neural underpinnings of links between language and action. Topics include neurocognitive studies of motor representations of speech sounds, action-related language, sign language and co-speech gestures. It will be concluded that there is strong evidence on the interaction between speech and gestures in the brain. This interaction however shares general properties with other domains in which there is interplay between language and action. PMID- 17416412 TI - High lubricious surface of cobalt-chromium-molybdenum alloy prepared by grafting poly(2-methacryloyloxyethyl phosphorylcholine). AB - Osteolysis caused by wear particles from polyethylene in artificial hip joints is of great concern. Various bearing couple combinations, bearing material improvements, and surface modifications have been attempted to reduce such wear particles. With the aim of reducing the wear and developing a novel artificial hip-joint system, we created a highly lubricious metal-bearing material: A 2 methacryloyloxyethyl phosphorylcholine (MPC) polymer was grafted onto the surface of the cobalt-chromium-molybdenum (Co-Cr-Mo) alloy. For ensuring the long-term retention of poly(MPC) on the Co-Cr-Mo alloy, we used a 4-methacryloxyethyl trimellitate anhydride (4-META) intermediate layer and photo-induced graft polymerization technique to create a strong bonding between the Co-Cr-Mo substrate and the poly(MPC) chain via the 4-META layer. The Co-Cr-Mo alloy was pretreated with nitric acid and O(2) plasma to facilitate efficient interaction between the 4-META carboxyl group and the surface hydroxyl group on the Cr oxide passive layer of the Co-Cr-Mo alloy. After MPC grafting, the MPC unit peaks were clearly observed in the Fourier-transform infrared spectroscopy with attenuated total reflection (FT-IR/ATR) and X-ray photoelectron spectroscopy (XPS) spectra of the Co-Cr-Mo surface. Tribological studies with a pin-on-plate machine revealed that surface MPC grafting markedly lowered the friction coefficient. We concluded that the grafted poly(MPC) layer successfully provided high lubricity to the Co-Cr-Mo surface. PMID- 17416413 TI - Clonogenic acute myelogenous leukemia cells are heterogeneous with regard to regulation of differentiation and effect of epigenetic pharmacological targeting. AB - Differentiation-inducing therapy with the DNA-methylation inhibitor Decitabine (5'-aza-deoxycytidine) and histone deacetylase (HDAC) inhibitors are now considered in acute myelogenous leukemia (AML). We investigated the in vitro effects of Decitabine and two structurally unrelated HDAC inhibitors (Sodium 4 phenyl butyrate, Tricostatin A) on clonogenic AML cells. Based on morphological criteria we identified four major colony types: (i) non-erythroid colonies, (ii) erythroid colonies that were detected only for a subset of patients and could be further sub classified into mature and immature forms, and (iii) intermediate colonies. Erythroid differentiation was associated with low CD34 expression. The colonies showed differences in morphology, viability, cell cycle distribution and expression of differentiation markers. Both Decitabine and the two HDAC inhibitors altered AML cell expression of differentiation markers, whereas the drugs did not have any major influence on cell cycle distribution. However, the pharmacological effects differed between the four colony subsets, and differences were also detected between the two HDAC inhibitors. We conclude that clonogenic AML cells can be classified into well-defined subsets based on their differentiation, and these subsets differ in their biological characteristics as well as their response to pharmacological targeting of epigenetic regulation. PMID- 17416414 TI - A new era for ribonucleoside reductase inhibition. PMID- 17416415 TI - Arsenic in the cerebrospinal fluid of a patient receiving arsenic trioxide for relapsed acute promyelocytic leukemia with CNS involvement. AB - We report on a 42-year-old patient whose relapse of acute promyelocytic leukaemia (APL) included meningeal infiltration. Since he had previously experienced ATRA syndrome, he received arsenic trioxide (ATO) plus intrathecal therapy with cytarabine, prednisone, and methotrexate. We measured the concentration of arsenic in his cerebrospinal fluid (CSF). Arsenic showed a peak CSF concentration of 0.008 mg/l (0.11 micromol/l) and a nadir of 0.002 mg/l (0.027 micromol/l), both representing about 14% of blood levels. ATO thus crosses the blood-CSF barrier when administered intravenously, but the concentration in CSF is probably not sufficient for treatment of meningeal leukemia. PMID- 17416416 TI - The incidence and survival of Waldenstrom's Macroglobulinaemia in South East England. AB - Waldenstrom's Macroglobulinaemia (WM) is an uncommon B-cell lymphoproliferative disorder defined as a predominately inter-trabecular bone marrow infiltration of small lymphocytes with an IgM monoclonal gammopathy. There are little reliable incidence and survival data for the disease in the UK since epidemiological studies have usually grouped it with other plasma cell dyscrasias. This study uses data from the South Thames Haematology Register and the Thames Cancer Registry for South East England to describe the incidence and survival of WM, and the influence of selected clinical factors on survival. Between 1999 and 2001, there were 152 new cases of WM recorded in the South Thames Haematology Register, giving an age standardised rate of 0.55 per 100,000 European standard population (0.73 for males and 0.42 for females). The incidence increased with age, and the median age at diagnosis was 75 years (range 45-93 years). The estimated 5 year survival was 57% (95% CI: 47-66%). Age over 70, haemoglobin less than 10 g/L and the Eastern Cooperative Oncology Group (ECOG) Performance Status grade 3-4 at diagnosis were associated with worse survival. Between 1985 and 2002, the Thames Cancer Registry recorded 750 cases of WM occurring in the wider area of South East England. The relative 5 year survival for patients aged less than 70 years was 70% (95% CI: 60-81%) and for patients aged 70 and over it was 50% (95% CI: 41 60%). PMID- 17416417 TI - Effects of anthocyanin-rich extract from red cabbage leaves on meristematic cells of Allium cepa L. roots treated with heavy metals. AB - The incubation of Allium cepa L. roots in Pb(NO3)2, Cd(NO3)(2)x4H2O or Cr(NO3)(3)x9H2O solution at the concentration of 100 microM lowered the mitotic index (MI) value in meristem by 58%, 39%, 48%, respectively. The proportion of mitotic phases (mainly prophases and telophases) in MI value was also changed. Moreover, mitotic disturbances such as: c-metaphases, sticky and lagging chromosomes, chromosome bridges, binucleate cells, micronuclei, "budding" nuclei and nucleoli partly outside nuclei were induced in the presence of the tested heavy metals, most frequently after Pb treatment. Pre-incubation in the ATH-rich extract from red cabbage leaves caused 2.5%, 1.8% or 1.6% increase in MI value as compared to the meristematic cells of A. cepa L. roots treated only with Cd, Pb, or Cr, respectively. Additionally, the ATH-rich extract was responsible for changing phase index values towards the control level in the material incubated in Pb or Cd. Moreover, the total number of mitotic abnormalities induced by the tested metals was reduced due to the preincubation in the ATH-rich extract, most effectively in the roots treated with Cd (by 2/3) while by half in Cr presence. These data suggest a protective action of the ATH-rich extract from red cabbage leaves against heavy metal toxicity. PMID- 17416418 TI - Animal experimentation in Japan: regulatory processes and application for microbiological studies. AB - We have conducted animal experimentation as a highly effective technique in biological studies. Also in microbiological studies, we have used experimentation to prevent and treat many infectious diseases in humans and animals. In Japan, the 'Law for the Humane Treatment and Management of Animals', which covers the consideration of the three R principles, refinement, replacement and reduction for an international humane approach to animal experimentation came into effect in June 2006. Looking towards the straightforward operation of the law in animal experimentation, three government ministries established new basic guidelines for experimentation performed in their jurisdictional research and testing facilities. For future microbiological studies involving animals in Japan, we need to perform animal experiments according to the basic guidelines in association with overseas management systems. In this report, we discussed essential actions for the management of animal experimentation in microbiological studies in Japan. PMID- 17416419 TI - A missense polymorphism in porcine interferon-gamma cDNA affects antiviral activity of the protein variant. AB - We determined the interferon-gamma (IFN-gamma) cDNA sequence from three porcine breeds, Duroc, Landrance/Duroc hybrid, and Landrance breeds. Five single nucleotide polymorphisms (SNPs) of porcine IFN-gamma (PoIFN-gamma) were identified, respectively, at positions 269 (A/G), 376 (C/T), 426 (T/C), and 465 (T/C) of the coding sequence in Landrance/Duroc hybrid, and at position 251 (A/G) in Landrance breed. Among them, A269G and A251G polymorphisms resulted in Q67R and K61R replacements in the mature protein. PoIFN-gamma cDNAs of Duroc breed (PoIFN-gamma-W) and Landrance/Duroc hybrid (PoIFN-gamma-M), which, respectively, encoded Q67 and R67, were introduced into a prokaryotic expression vector pET32 to express recombinant PoIFN-gamma-W (rPoIFN-gamma-W) and rPoIFN-gamma-M protein variants in Escherichia coli. The identity of both protein variants was further confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We then compared bioactivities of these two recombinant proteins. Although both recombinant protein variants exhibited comparable activities in antiproliferation of PK-15 cells and in nitric oxide (NO) induction of porcine peripheral monocytes, antiviral activity of rPoIFN gamma-W protein was significantly higher (P<0.001) than that of rPoIFN-gamma-M protein in a plaque inhibition assay using pseudorabies virus (PRV). IC50 values of rPoIFN-gamma-W and rPoIFN-gamma-M protein in anti-PRV assay were determined as 5.3+/-1.3 and 9.3+/-4.3nM, respectively. In conclusion, we have identified five novel SNPs in PoIFN-gamma cDNA, including two missense polymorphisms that result in Q67R and K61R replacements. Our results further demonstrate that Q67R can markedly reduce antiviral activity of the PoIFN-gamma protein. This is the first report that shows the functional SNP in the coding region of IFN-gamma. In the future, it is imperative to determine whether Q67R replacement in IFN-gamma may have disease association. PMID- 17416420 TI - Intrinsic and extrinsic factors associated with night waking in 9-month-old infants. AB - This study examined relations between infant night waking and both daytime behaviors reflective of poor behavioral and emotional regulation (intrinsic factors) and parent behaviors that may contribute to infant night waking (extrinsic factors) in 41 infants. Mothers completed questionnaires and an infant sleep and crying diary. More time awake at night was related to separation distress, frequent daytime crying, dysregulation, co-sleeping with parents, breast feeding, and being put to bed asleep. More frequent waking was related to separation distress, frequent daytime crying, co-sleeping, and breast feeding. The combination of intrinsic and extrinsic factors predicted night waking better than behaviors from one category alone. Implications for parenting are discussed. PMID- 17416421 TI - The effect of positive emotion on infants' gaze shift. AB - Infants often voluntarily glance at their social partner during their toy play, disengaging their gaze from a toy and selecting a caregiver as their new looking target. This study posed two research questions: Do positive emotions disengage infants' gaze from their point of fixation, and do positive emotions facilitate the selection of the caregiver as their next looking target? The rate of gaze shifts was calculated for neutral and positive emotional states during their toy play. Across all ages, infants exhibited more disengagement from their point of fixation in the positive state than in the neutral one. However, 6- and 9-month old infants revealed no difference in selecting a caregiver or a physical object as their next looking target in the positive state, but 12-month-olds increased gazing at caregivers in the positive state. These results were discussed with regard to the role of positive emotions on the development of infants' initiating joint attention. PMID- 17416422 TI - Communication and crying in newborns. AB - Few studies have analysed newborns' crying behaviour in relation to the communicative context. In the present study, duration, latency and dysphony of newborns' cry were analysed in three different communicative conditions: absent, continuous and discontinuous tactile communication. Thirty newborns were randomly assigned to three tactile communicative procedures. Subsequently they were presented with two visual stimuli, face (social-stimulus) and square (non-social stimulus). The study showed that newborns cried only during visual stimuli presentation both in absent and continuous communication conditions. Newborns with absent communication cried more, with shorter latency and more dysphony than newborns with continuous communication. No significant differences were found in duration, latency and dysphony of newborns' cry between the two stimuli. Newborns in the discontinuous communicative condition did not cry either during tactile communication or during visual stimuli presentation. Different communicative contexts affected the newborns' crying behaviour during stimuli presentation. Different interpretations are discussed here in order to explain the results. PMID- 17416423 TI - CO2 laser supraglottoplasty for severe laryngomalacia: a study of symptomatic improvement. AB - OBJECTIVE: To investigate post-operative symptom improvement in patients with severe laryngomalacia. STUDY DESIGN: Severe laryngomalacia was diagnosed in 138 patients (average age of 6.97 months) by bronchoscopy. Laryngomalacia was defined by the direction of supraglottic collapse: type A (posterolateral), type B (complete), and type C (anterior). As multiple laryngomalacia types within an individual were common, patients were further categorized into group I (type A only), group II (type B or B+A), and group III (type C, C+A, or C+B+A). CO(2) laser supraglottoplasty was performed. Improvements in inspiratory stridor, suprasternal retraction, substernal retraction, feeding difficulty, choking, post feeding vomit, failure to thrive, and cyanosis were investigated. The presence of a symptom was scored as 1, and the absence as 0. The total score of symptoms was calculated for each patient. General medical history, age at time of surgery, type of laryngomalacia, post-operative intubation period, duration in ICU and dates of postoperative admission were recorded. RESULTS: Overall symptom improvement was observed in 82.6% of patients, with statistically significant resolution evident in group III (B-value=0.79, 95% CI: -0.01, 1.59). Symptoms were not well improved in patients with cerebral palsy (n=32, B-value=-1.02, 95% CI: -1.80, -0.25; p<0.01). The two most improved symptoms were substernal retraction and suprasternal retraction, while the two least improved symptoms were choking and feeding difficulties. CONCLUSION: CO(2) laser supraglottoplasty is an effective treatment option for severe laryngomalacia, especially for group III laryngomalacia cases in the absence of cerebral palsy. It has the superiority of facilitating significant symptomatic resolution and reducing the post operative complications. PMID- 17416424 TI - PPAR-gamma: therapeutic target for ischemic stroke. AB - The peroxisome proliferator activated receptors (PPARs), which belong to the nuclear receptor superfamily, are key regulators of glucose and fat metabolism. The PPAR-gamma isoform is involved in the regulation of cellular glucose uptake, protection against atherosclerosis and control of immune reactions. In addition, the activation of PPAR-gamma effectively attenuates neurodegenerative and inflammatory processes in the brain. Here, we review a novel aspect of beneficial and clinically relevant PPAR-gamma actions: neuroprotection against ischemic injury mediated by intracerebral PPAR-gamma, which is expressed in neurons and microglia. Together with the recent observation that the PPAR-gamma ligand pioglitazone reduces the incidence of stroke in patients with type 2 diabetes, this review supports the concept that activators of PPAR-gamma are effective drugs against ischemic injury. PMID- 17416425 TI - A short history of the rise of the molecular pharmacology of ionotropic drug receptors. AB - Remarkably, perhaps, for many pharmacologists today, just over 25 years ago, receptors were still considered hypothetical entities. The isolation and identification of Langley's receptive substance (Ehrlich's side-chains) required efforts from diverse groups; serendipity also facilitated its purification and subsequent biochemical and molecular characterization. In this review, I consider some of the key individuals and breakthrough technical developments from the late 1950s to the early 1990s that lead to the cloning of the first receptors. I focus on the nicotinic acetylcholine receptor to illustrate the complexities in this field and because it was the first receptor to be cloned. This brief history will also touch upon the implications of the rise of molecular pharmacology for the development of new drugs. PMID- 17416426 TI - Restoring HOmeostasis: is heme oxygenase-1 ready for the clinic? AB - Inflammation and immunity result in a wide range of disease processes, including atherosclerosis, vascular thrombosis and sepsis. Heme oxygenase-1 (HO-1) is a key enzyme that is integral to the temporal and spatial regulation of the host response and, together with its products carbon monoxide (CO) and bilirubin, is crucial for maintaining homeostasis and the preservation of function and life. An increasing number of reports demonstrates that HO-1, CO and bilirubin regulate the immune response. As CO and bilirubin enter clinical trials, there are obstacles to be addressed before their full therapeutic potential can be achieved. In this article, we delineate the challenges that lie ahead regarding toxicity, pharmacokinetics and mechanisms of action to be able to take full advantage of the powerful cytoprotective properties of these agents for clinical benefit. PMID- 17416427 TI - Mutation analysis of lamivudine resistant hepatitis B virus strains by TaqMan PCR. AB - Hepatitis B virus infected patients on long-term lamivudine treatment are exposed to a 15-32% risk compounded annually of developing resistance mutations. Such resistance results in a progression of the liver damage caused by chronic hepatitis B, and may also impair the effect of other antivirals through cross resistance. At present lamivudine is used frequently as monotherapy because of its relatively low price and negligible side effects. Thus, simple methods for identifying resistance mutations are required. A method based on real-time polymerase chain reaction with TaqMan chemistry is described. The method combines both primer specificity, in order to target wild type and mutant viral strains at codon 180, and a mixture of three minor groove binding probes distinguishing the YMDD wild type and the YVDD and YIDD variants at codon 204. The accuracy of the method was verified by concordance with results of direct sequencing and restriction fragment length polymorphism when examining 27 samples from five patients, in whom lamivudine resistance was known to have developed. This method is rapid, cost effective, and should prove useful for monitoring patients treated with lamivudine. PMID- 17416428 TI - High throughput functional analysis of HIV-1 env genes without cloning. AB - Functional human immunodeficiency virus type 1 (HIV-1) env genes have been widely used for vaccine design, neutralization assays, and pathogenesis studies. However, obtaining bona fide functional env clones is a time consuming and labor intensive process. A new high throughput method has been developed to characterize HIV-1 env genes. Multiple rev/env gene cassettes were obtained from each of seven HIV-1 strains using single genome amplification (SGA) PCR. The cytomegalovirus (CMV) promoter was amplified separately by PCR. A promoter PCR (pPCR) method was developed to link both PCR products using an overlapping PCR method. Pseudovirions were generated by cotransfection of pPCR products and pSG3 Delta env backbone into 293T cells. After infecting TZM-bl cells, 75 out of 87 (86%) of the rev/env gene cassettes were functional. Pseudoviruses generated with pPCR products or corresponding plasmid DNA showed similar sensitivity to six HIV 1 positive sera and three monoclonal antibodies, suggesting neutralization properties are not altered in pPCR pseudovirions. Furthermore, sufficient amounts of pseudovirions can be obtained for a large number of neutralization assays. The new pPCR method eliminates cloning, transformation, and plasmid DNA preparation steps in the generation of HIV-1 pseudovirions. This allows for quick analysis of multiple env genes from HIV-1 infected individuals. PMID- 17416429 TI - Insertions in the beta3-beta4 loop of reverse transcriptase of human immunodeficiency virus type 1 and their mechanism of action, influence on drug susceptibility and viral replication capacity. AB - Introduction of antiretroviral therapy combining protease and reverse transcriptase (RT) inhibitors has dramatically improved the quality of life and survival of patients infected with the human immunodeficiency virus (HIV). However, effective long-term therapy of HIV-infection has been severely hampered by the development of drug resistance. Resistance to antiretroviral drugs is generally conferred by specific amino acid substitutions in the target gene of the drug. Yet, occasionally gene insertions are being observed. The most commonly observed insertion is seen during substrate analogue RT inhibitor therapy and is selected in the beta3-beta4 loop of the RT enzyme. This flexible loop is located in the fingers subdomain of the enzyme and plays an important role in substrate binding. The acquisition of drug resistance related mutations or insertions might come at a price, which is reduced performance of the enzyme resulting in a diminished replication capacity of the virus. Various types of insertions have been described, and, in this review, we have summarized these data and discussed the mechanism of action of the RT inserts and their impact on both drug susceptibility and replication capacity. PMID- 17416430 TI - The effect of veterinary-treated clinical mastitis and pregnancy status on culling in Swedish dairy cows. AB - The interaction of the effects of pregnancy status and veterinary-treated clinical mastitis on culling in Swedish dairy cattle was analyzed with survival analysis. The data were from 978,780 cows with first calvings between 1988 and 1996. Four breeds (Swedish Red and White (SRB), Swedish Friesian (SLB), Swedish Polled Breed and Jersey) were included in the analysis, together with the SRB x SLB crossbreds. Length of productive life was defined as the number of days between first calving and culling or censoring (end of data collection). The model (Weibull proportional hazard) included the interaction of parity by pregnancy status by veterinary-treated clinical mastitis, peak test-day milk yield deviation within herd-year-parity, age at first calving, year by season, region, breed, herd production level, and the random effect of herd. The effects of pregnancy status and veterinary-treated clinical mastitis were modeled as time dependent covariates. The lactation was divided into five stages during which a veterinary-treated clinical mastitis and culling might occur and in which the pregnancy status was assumed to be known and culling could occur. Open cows had a pronounced effect on culling: they had a very high risk of being culled in all lactations, and it was even higher if they were treated for mastitis in early lactation. For pregnant cows, the later they got pregnant during the lactation, the greater their risk to be culled. The risk associated with cases of veterinary treated clinical mastitis remained important throughout the lactation. PMID- 17416431 TI - Dystroglycan: a possible mediator for reducing congenital muscular dystrophy? AB - Alpha-dystroglycan is a highly glycosylated peripheral protein forming a complex with the membrane-spanning beta-dystroglycan and establishing a connection between the extracellular matrix and the cytoskeleton. In skeletal muscle, as part of the larger dystrophin-glycoprotein complex, dystroglycan is believed to be essential for maintaining the structural and functional stability of muscle fibers. Recent work highlights the role of abnormal dystroglycan glycosylation at the basis of glycosyltransferase-deficient congenital muscular dystrophies. Notably, modulation of glycosyltransferase activity can restore alpha dystroglycan receptor function in these disorders. Moreover, transgenic approaches favoring the interaction between dystroglycan and the extracellular matrix molecules also represent an innovative way to restore skeletal muscle structure. These pioneering approaches might comprise an important first step towards the design of gene-transfer-based strategies for the rescue of congenital muscular dystrophies involving dystroglycan. PMID- 17416432 TI - Production of poly-beta-hydroxybutyrate (PHB) by Vibrio spp. isolated from marine environment. AB - Bacteria isolated from marine sediments were screened for their ability to accumulate polyhydroxyalkanoates. Among the isolates, four Vibrio spp. (strain M11, M14, M20 and M31) were studied in detail. All synthesized intracellular lipid inclusions during growth on diverse carbon sources including acetate, glycerol, succinate, glucose and sucrose. The inclusions were identified to be poly-beta-hydroxybutyrate (PHB) using gas chromatography and nuclear magnetic resonance analysis. No other type of polyhydroxyalkanoates (PHAs) was found to be accumulated by these marine isolates, suggesting that the diversity of PHAs produced in marine environments may be not as versatile as found in other environments. Strain M11 accumulated PHB in concentrations as high as 41% of cell dry weight when grown in medium containing 4% of sodium chloride. One of the Vibrio spp. was identified to be closely related to Vibrio natriegens (98% identity) by partial 16S rDNA sequence homology. V. natriegens has the shortest generation time (9.8 min) of any bacterium and this characteristic may be an exploitable trait for the industrial production of PHB. PMID- 17416433 TI - Dengue viruses can infect human primary lung epithelia as well as lung carcinoma cells, and can also induce the secretion of IL-6 and RANTES. AB - Dengue viruses (DENV) are herein demonstrated for the first time as being able to infect and replicate in human primary lung epithelium and various lung cancer cell lines. The detection of dengue virus particles and viral negative strand RNA synthesis in the cell, in conjunction with the release of viral progenies in culture supernatants, support the notion that lung cells are susceptible to dengue virus infection. The replication efficiency of DENV in lung cancer cells from high to low is: DEN-2 (dengue virus type-2), DEN-3, DEN-4 and DEN-1. Moreover, the susceptibility of the six lung cancer cell lines to DEN-2 infection is: SW1573>A549>H1435; H23; H520; Bes2B. DEN-2 infection significantly increased the expression levels of IL-6 and RANTES in four of the six lung cancer cell lines, which is consistent with the high expression levels of these molecules in DHF/DSS patients. IL-6 expression induced by DEN-2 infection was NF-kappaB dependent. In summary, our results indicate that lung epithelial cell is a possible target of dengue viruses and IL-6 and RANTES may play pivotal roles in lung related immuno-pathogenesis. PMID- 17416435 TI - Monoclonal antibody and porcine antisera recognized B-cell epitopes of Nsp2 protein of a Chinese strain of porcine reproductive and respiratory syndrome virus. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important pathogens for swine industry. The non-structural protein 2 (Nsp2) is considered to be one of the immunogenic proteins of PRRSV. In this study, the B-cell epitopes of the Nsp2 protein of a North American type Chinese strain PRRSV BJ-4 were identified on a prokaryotic expressed Nsp2 fragment (73 567aa). A total of six monoclonal antibodies (mAbs) recognizing different epitopes on the expressed protein were prepared. All six mAbs exhibited immunoreactivity with the denatured Nsp2 protein in Western blotting and produced strong perinuclear staining in PRRSV infected MARC-145 cells in an immunofluorescence assay. Pepscan analysis revealed six distinct linear epitopes for the six mAbs, respectively, and of which four were identified to be novel linear Nsp2 B-cell epitopes: T(73)LPERVRPPDDWAT(86), D(385)ELKDQMEED(394), P(452)VPAPRRKVGSDCGS(466), and P(467)VSLGGDVPNS(477). All of the six mAb specific peptides could be recognized by porcine PRRSV antiserum, indicating that the epitopes involving these synthetic peptides were immunogenic and immunodominant during PRRSV infection in pigs. Our results provided valuable information for developing novel PRRSV vaccines using the Nsp2 epitopes as potential serological markers. PMID- 17416436 TI - Perturbation of erythrocyte antioxidant barrier, lipid peroxidation and protein carbonylation in non-diabetic first degree relatives of patients with type 2 diabetes. AB - OBJECTIVE: Oxidative stress plays an important role in the pathogenesis of type 2 diabetes. But it is still discussed whether oxidative stress precedes or merely reflects diabetic complications. The present study was carried out to search for the possibility of oxidative stress among the first degree relatives of patients with type 2 diabetes, as they are more prone to develop type 2 diabetes. METHODS: This study has been conducted on 30 first degree relatives of patients with type 2 diabetes and 34 healthy subjects without any known family history of diabetes. Whole blood glutathione, plasma malondialdehyde (MDA), protein carbonylation, fasting glucose levels and the activities of anti-oxidant enzymes glutathione peroxidase, catalase and glutathione S-transferase were measured. RESULTS: The antioxidant enzyme glutathione peroxidase, plasma MDA and protein carbonyl levels were significantly elevated in the test group compared with controls. The glutathione levels were significantly decreased in the test group. CONCLUSION: This study reveals alteration of antioxidant status and oxidative stress among the first degree relatives of patients with type 2 diabetes. PMID- 17416434 TI - Vaccines to prevent severe acute respiratory syndrome coronavirus-induced disease. AB - An important effort has been performed after the emergence of severe acute respiratory syndrome (SARS) epidemic in 2003 to diagnose and prevent virus spreading. Several types of vaccines have been developed including inactivated viruses, subunit vaccines, virus-like particles (VLPs), DNA vaccines, heterologous expression systems, and vaccines derived from SARS-CoV genome by reverse genetics. This review describes several aspects essential to develop SARS CoV vaccines, such as the correlates of protection, virus serotypes, vaccination side effects, and bio-safeguards that can be engineered into recombinant vaccine approaches based on the SARS-CoV genome. The production of effective and safe vaccines to prevent SARS has led to the development of promising vaccine candidates, in contrast to the design of vaccines for other coronaviruses, that in general has been less successful. After preclinical trials in animal models, efficacy and safety evaluation of the most promising vaccine candidates described has to be performed in humans. PMID- 17416437 TI - Changes and factors associated with dentists' willingness to treat patients with severe disabilities. AB - OBJECTIVE: This study investigated changes in dentists' willingness to treat severely disabled patients and to understand dentists' opinions on reimbursements after the implementation of a dental care financial reward program in Taiwan. METHODS: Three hundred dentists from 29 teaching hospitals were randomly selected to answer a structured questionnaire, and 184 structured questionnaires were returned. Multiple regression analysis was used to examine the factors associated with dentists' willingness to treat severely disabled patients. RESULTS: Approximately 60% of the dentists said reimbursements for treatment of severely disabled patients were reasonable. 50.4% of dentists were willing or very willing to treat disabled patients. Seventy-nine percent dentists affected by the program had a higher willingness but 83.7% dentists said this program did not make a significant difference to their income. 52.8% of dentists agreed the program would increase the quality of dental care. The factors significantly affecting dentists' willingness included dentist's age, specialty field, perception of the program in promoting the quality of dental services, and perception of the ability to provide adequate treatments for severely disabled patients. CONCLUSIONS: The rewards program significantly increased the willingness of most hospital-base dentists to treat the severely disabled patients although the effect of incentive to their income was limited. PMID- 17416438 TI - Beyond ecological traps: perceptual errors and undervalued resources. AB - Ecological traps arise when animals choose to settle in poor-quality habitats, leading to a reduction in their survival or productivity. Although this phenomenon has received comprehensive theoretical treatment in the recent literature, the corollary of the ecological trap (when animals choose to avoid good-quality habitats) is rarely discussed. Failure to recognize high-quality sites could influence the ability of a population to reach its threshold size. An exploration of this phenomenon brings new insights into existing cases of ecological traps. By considering the full range of pitfalls faced during settlement, the relationship between habitat quality and population size might become clearer. We argue here that an improved understanding of factors determining the colonization of high-quality sites could aid conservationists in mitigating the damaging effects of maladaptive habitat selection. PMID- 17416439 TI - Phenotype ontologies: the bridge between genomics and evolution. AB - Understanding the developmental and genetic underpinnings of particular evolutionary changes has been hindered by inadequate databases of evolutionary anatomy and by the lack of a computational approach to identify underlying candidate genes and regulators. By contrast, model organism studies have been enhanced by ontologies shared among genomic databases. Here, we suggest that evolutionary and genomics databases can be developed to exchange and use information through shared phenotype and anatomy ontologies. This would facilitate computing on evolutionary questions pertaining to the genetic basis of evolutionary change, the genetic and developmental bases of correlated characters and independent evolution, biomedical parallels to evolutionary change, and the ecological and paleontological correlates of particular types of change in genes, gene networks and developmental pathways. PMID- 17416440 TI - A meta-analysis to assess the efficacy of oral antiviral treatment to prevent genital herpes outbreaks. AB - BACKGROUND: Efficacy of oral antiviral therapies, ie, acyclovir, valacyclovir (VACV), and famciclovir, for suppression of recurrent genital herpes was studied at different doses and regimens. OBJECTIVE: We sought to compare the clinical efficacies of the different oral antiviral drugs prescribed prophylactically to suppress recurrent genital herpes. METHODS: MEDLINE and EMBASE databases were searched for articles on genital herpes and selected antiviral drugs. The selected trials were: parallel randomized clinical trials testing prophylactic oral antiviral treatment of genital herpes versus placebo in immunocompetent and nonpregnant patients. RESULTS: Fourteen randomized clinical trials were selected, including a total of 6158 patients. The global relative risk of developing at least one recurrence during the study was reduced by 47% (95% confidence interval 45%-49%) in antiviral drug groups compared with the placebo. The best evaluated regimens, with comparable efficacies, were given twice daily, ie, acyclovir (400 mg twice daily), VACV (250 mg twice daily), and famciclovir (250 mg twice daily), or once daily (VACV 500 mg). LIMITATIONS: The only end point available for all the studies was the number of patients presenting at least one recurrence of genital herpes during the observation period. CONCLUSION: The results of this first meta-analysis confirmed the high clinical efficacy of oral acyclovir, VACV, or famciclovir for prophylaxis against recurrent genital herpes. PMID- 17416441 TI - Creatine improves health and survival of mice. AB - The supplementation of creatine (Cr) has a marked neuroprotective effect in mouse models of neurodegenerative diseases. This has been assigned to the known bioenergetic, anti-apoptotic, anti-excitotoxic, and anti-oxidant properties of Cr. As aging and neurodegeneration share pathophysiological pathways, we investigated the effect of oral Cr supplementation on aging in 162 aged C57Bl/6J mice. Outcome variables included "healthy" life span, neurobehavioral phenotyping, as well as morphology, biochemistry, and expression profiling from brain. The median healthy life span of Cr-fed mice was 9% higher than in control mice, and they performed significantly better in neurobehavioral tests. In brains of Cr-treated mice, there was a trend towards a reduction of reactive oxygen species and significantly lower accumulation of the "aging pigment" lipofuscin. Expression profiling showed an upregulation of genes implicated in neuronal growth, neuroprotection, and learning. These data show that Cr improves health and longevity in mice. Cr may be a promising food supplement to promote healthy human aging. PMID- 17416442 TI - An atom counting strategy towards analyzing the biological activity of sex hormones. AB - A simple and effective molecular descriptor, viz., the number of atoms in a molecule (N(A)) is made use of in the development of the quantitative structure activity relationship (QSAR). A series of testosterone derivatives with various biological activities and estrogen derivatives with the activities in terms of relative binding affinity (RBA) are considered to find out the potential of N(A) in predicting the activities of those molecules. It is heartening to note that N(A) along with the electrophilicity index (omega) is capable of explaining the biological activities of the male and female hormones. PMID- 17416443 TI - Manipulation of the infectious bronchitis coronavirus genome for vaccine development and analysis of the accessory proteins. AB - Infectious bronchitis coronavirus (IBV) is the cause of the single most economically costly infectious disease of domestic fowl in the UK--and probably so in many countries that have a developed poultry industry. A major reason for its continued dominance is its existence as many serotypes, determined by the surface spike protein (S), cross-protection being poor. Although controlled to some degree by live and inactivated vaccines, a new generation of IB vaccines is called for. Reverse genetic or 'infectious clone' systems, which allow the manipulation of the IBV genome, are key to this development. New vaccines would ideally be: genetically stable (i.e. maintain a stable attenuated phenotype); administered in ovo; and be flexible with respect to the source of the spike protein gene. Rational attenuation of IBV requires the identification of genes that are simultaneously not essential for replication and whose absence would reduce pathogenicity. Being able to modify a 'core' vaccine strain to make it applicable to a prevailing serotype requires a procedure for doing so, and the demonstration that 'spike-swapping' is sufficient to induce good immunity. We have demonstrated that four small IBV proteins, encoded by genes 3 and 5, are not essential for replication; failure to produce these proteins had little detrimental affect on the titre of virus produced. Our current molecularly cloned IBV, strain Beaudette, is non-pathogenic, so we do not know what effect the absence of these proteins would have on pathogenicity. That said, plaque size and composition of various gene 3/5 recombinant IBVs in cell culture, and reduced output and ciliostasis in tracheal organ cultures, shows that they are less aggressive than the wild-type Beaudette. Consequently these genes remain targets for rational attenuation. We have recently obtained evidence that one or more of the 15 proteins encoded by gene 1 are also determinants of pathogenicity. Hence gene 1 is also a target for rational attenuation. Replacing the S protein gene of Beaudette with that from the pathogenic M41 strain resulted in a recombinant virus that was still non-pathogenic but which did induce protection against challenge with M41. We have since made other 'spike-swapped' recombinants, including ones with chimaera S genes. Uniquely, our molecular clone of Beaudette is benign when administered to 18-day-old embryos, even at high doses, and induces immunity after this route of vaccination. Taken together, our results point to the creation of a new generation of IB vaccines, based on rational modification of the genome, as being a realisable objective. PMID- 17416444 TI - Benign schwannoma of the left cervical phrenic nerve. AB - A patient presented with a benign schwannoma of the cervical phrenic nerve on the left side of the neck. Analysis of the aspirate suggested the diagnosis and it was confirmed on imaging. The patient had the lesion excised with preservation of the phrenic nerve. PMID- 17416446 TI - Benzo[a]pyrene-induced elevation of GSH level protects against oxidative stress and enhances xenobiotic detoxification in human HepG2 cells. AB - Glutathione (GSH) is one of the most important antioxidants in mammalian cells. It also plays an important role in chemical detoxification. Some evidence showed that polycyclic aromatic hydrocarbons, such as benzo[a]pyrene (B[a]P [50-32-8]), could increase GSH content as a defense mechanism against oxidative stress as well as to promote its detoxification. However, there has been very little study on clarifying the role GSH plays in antioxidation and detoxification actions. Therefore, the present study aims to analyze intracellular glutathione metabolism in the human hepatoma cells (HepG2) upon exposure to B[a]P. Exposure of the cells to B[a]P (1-100 microM) for 24 h did not cause significant cell death in this cell line. By selecting the sublethal concentration of 10 microM, B[a]P caused a significant increase in GSH and a small (13%) but significant decrease in glutathione reductase activity. However, there was no change in the activity of glutathione peroxidase, and no detectable increase in reactive oxygen species (ROS) production. Treatment with B[a]P caused up to 1.5 folds increase in gamma glutamylcysteine synthatase (gamma-GCS) activity over control. Buthioneine sulfoximine (BSO), an inhibitor of gamma-GCS, could suppress GSH increase in a dose-dependent manner. Assessment of the oxidative state of the cells indicated that the increase in GSH caused the cells to become more reduced. Thus, the results concluded that cells were not suffering from oxidative stress at 24 h after treatment with 10 microM B[a]P. Upon analyzing the activities of detoxification enzymes, there was an increase in the activity of CYP1A subfamily monooxygenases and glutathione S-transferase. Both changes occurred prior to the changes in gamma-GCS activity and the increase in GSH. In summary, results of the present study demonstrate that B[a]P caused an activation of detoxification enzymes. The increase in intracellular GSH level was due to activation of gamma GCS activities. Oxidative stress may not be an important risk factor for B[a]P (at 10 microM of up to 24 h) induced injury. PMID- 17416447 TI - Effect of hypoxia on RAW264.7 macrophages apoptosis and signaling. AB - Hypoxia, a decrease in oxygen tension occurring in pathological tissues, has a profound effect on macrophage functions. In this study, we have investigated cellular and molecular responses of murine macrophage-like RAW264.7 cells to low oxygen tension. Our study revealed that hypoxia induced RAW264.7 cells apoptosis and cell cycle arrest at G0/G1 phase. The result of Western blotting showed that the expression of apoptosis related signaling molecules, such as AKT and JNK was activated under hypoxia. The result of electrophoretic mobility shift assay showed that the DNA binding activity of nuclear factor kappa B (NF-kappaB) was also increased by hypoxic stimulation. Furthermore, gene expression profiles of RAW264.7 macrophages induced by hypoxia showed that hypoxia treatment may alter expression of genes related to apoptosis, survival, cell cycle, metabolism and structural matrix. PMID- 17416448 TI - Acute myocardial infarction complicated by ventricular standstill terminated by thrombolysis and transcutaneous pacing. AB - Although the overall need for emergency pacing following AMI has reduced with the wide spread use of thrombolysis the availability of transcutaneous pacing offers an emergency non-invasive strategy to optimise circulation following CHB or ventricular standstill. Transcutaneous pacing also facilitates the safe instigation of thrombolytic therapy to achieve reperfusion of the conduction system following AMI. PMID- 17416449 TI - Near death experiences, cognitive function and psychological outcomes of surviving cardiac arrest. AB - Cardiac arrest is associated with a number of cognitive processes as well as long term psychological outcomes. Recent studies have indicated that approximately 10 20% of cardiac arrest survivors report cognitive processes, including the ability to recall specific details of their resuscitation from the period of cardiac arrest. In addition it has been demonstrated that these cognitive processes are consistent with the previously described near death experience and that those who have these experiences are left with long term positive life enhancing effects. There have also been numerous studies that have indicated that although the quality of life for cardiac arrest survivors is generally good, some are left with long term cognitive impairments as well as psychological sequelae such as post-traumatic stress disorder. This paper will review near death experiences, cognitive function and psychological outcomes in survivors of cardiac arrest. PMID- 17416451 TI - Editorial comment on: A prospective randomized study comparing monopolar and bipolar transurethral resection of prostate using transurethral resection in saline (TURIS) system. PMID- 17416452 TI - Editorial comment on: A prospective randomized study comparing monopolar and bipolar transurethral resection of prostate using transurethral resection in saline (TURIS) system. PMID- 17416453 TI - A prospective randomized study comparing monopolar and bipolar transurethral resection of prostate using transurethral resection in saline (TURIS) system. AB - OBJECTIVES: To compare transurethral resection of prostate (TURP) using monopolar and bipolar transurethral resection in saline (TURIS) system. MATERIALS AND METHODS: A prospectively randomized study was conducted between January 2004 and January 2005. Patient demographics and indications for surgery were recorded. The safety end points studied were occurrence of complications and decline in postoperative serum sodium (Na(+)) and hemoglobin (Hb) levels. Efficacy end points were resection time, weight of resected prostate tissue, and improvement in International Prostate Symptoms Score (IPSS) and maximum flow rate (Q(max)) in patients' uroflow over 12 mo. RESULTS: One hundred consecutive patients were randomized and completed the study, with 52 patients in the monopolar TURP group and 48 in the TURIS group. At baseline, the two groups were comparable; they had at least 12 mo of follow-up. Mean resection time and mean weight of resected prostate tissue were comparable for both groups. Declines in the mean postoperative serum Na(+) for TURIS and monopolar TURP groups were 3.2 and 10.7 mmol/l, respectively (p<0.01). However, there was no statistical difference in the decline in postoperative Hb between the two groups. There were two cases of clinically significant transurethral resection syndrome in the monopolar group. Urethral strictures were observed in three cases of TURIS and one patient in the monopolar group. The IPSS and Q(max) improvements were comparable between the two groups at 12 mo of follow-up. CONCLUSIONS: Bipolar TURP using the TURIS system is clinically comparable to monopolar TURP at 1 yr with an improved safety profile. PMID- 17416454 TI - FISH: a tool we really need? PMID- 17416455 TI - Re: Fabio Calabro and Cora N. Sternberg. Current indications for chemotherapy in prostate cancer patients. Eur urol 2007;51:17-26. PMID- 17416456 TI - The long-term outcome of medical therapy for BPH. AB - OBJECTIVES: The lack of cure with medical therapy implies life-long treatment emphasising the need for a thorough understanding of the long-term outcome. We review the natural history, markers for progression, placebo effect, efficacy, pharmacoeconomic aspects, and preventive measures. METHODS: Literature review with particular reference to long-term controlled studies using plant extracts, alpha1-blockers, 5alpha-reductase inhibitors (5-ARIs), and combination therapy. RESULTS: There is a long-lasting (>or=12 mo) placebo response of symptoms (20% decrease) and maximum flow rate (10% rise). The five long-term controlled trials of plant extracts are inconclusive and therefore their role in contemporary medical management is still controversial. The alpha1-blockers provide fast amelioration of symptoms yet have no relevant impact on the risk of acute urinary retention or surgery. Combination therapy should be reserved for moderately or severely symptomatic patients with a high risk of progression; in the majority of patients the alpha1-blocker can be safely stopped after 6-12 mo. The preventive use of 5-ARIs in men with no or mild symptoms at risk of progression is scientifically sound yet not generally accepted mainly for economic reasons. CONCLUSIONS: A sharp contrast exists between the duration of the longest controlled trial (4.5 yr) and the situation in real life with treatment periods up to one or two decades of life. Real-life and registry data will be the only source of this important information in the future. PMID- 17416457 TI - A role of FRK in regulation of embryonal pancreatic beta cell formation. AB - The fyn-related-kinase (FRK) is a non-receptor tyrosine kinase expressed in various tissues, and among them, is the islets of Langerhans. The role of FRK in pancreatic beta cells has been addressed by studies of knockout or FRK transgenic mice. These experiments have shown that FRK overexpression in beta cells leads to an increased susceptibility to the beta cell toxin streptozotocin and to cytotoxic cytokines, suggesting that FRK may participate in events leading to beta cell destruction. However, these mice also exhibit an increased relative beta cell volume and increased beta cell replication following partial pancreatectomy, suggesting a positive role for FRK in the regulation of beta cell number as well. To further assess the significance of FRK for beta cell replication, we studied the beta cell area and islet cell replication in FRK null mice. We currently observed that the FRK knockout mouse showed no difference in the insulin positive cell area or in the percentage of Ki67-stained proliferating islet cells at adulthood, when compared to wild-type control. In addition, adult FRK(-/-) mice performed normally when subjected to an intravenous glucose tolerance test. To elucidate whether FRK affects pancreatic beta cell number during embryogenesis and shortly after birth, pancreata were collected from FRK( /-) mice at these stages. Histological analysis of insulin stained pancreatic sections showed that the insulin positive cell area in FRK(-/-) mice was reduced at embryonal day 15 and at birth to 31 and 70% of that of wild-type mice, respectively. FRK(-/-) pancreas weight on day 1 neonatally was similar to that of the control, indicating that the obtained results were not due to altered pancreatic growth. Taken together, these results show that FRK affects beta cell number during embryogenesis and early in life, but is probably redundant for beta cell number and function in adult animals under normal conditions. PMID- 17416458 TI - Activation of protein kinase Czeta mediates luteinizing hormone- or forskolin induced NGFI-B expression in preovulatory granulosa cells of rat ovary. AB - We have previously demonstrated that luteinizing hormone (LH) induces a rapid and transient expression of NGFI-B in the ovary. In this report, we investigated the signaling pathway for LH- and forskolin-induced NGFI-B expression in cultured rat granulosa cells of preovulatory follicles. LH- or forskolin-induced NGFI-B expression was suppressed by high dose of protein kinase C (PKC) inhibitor RO 31 8220 (10 microM), but not by low doses RO 31-8220 (0.1-1.0 microM) or adenylate cyclase inhibitor MDL-12,300A, implicating the involvement of atypical PKCs. Kinase assay revealed that LH treatment of granulosa cells resulted in a rapid stimulation of atypical PKCzeta activity. Interestingly, like LH, forskolin was also able to activate PKCzeta. Treatment with the cell-permeable PKCzeta-specific inhibitor pseudosubstrate peptide inhibited LH-or forskolin-induced NGFI-B expression, indicating the essential role of PKCzeta. Consistent with this promise, in granulosa cells depleted of diacylglycerol sensitive PKCs by prolonged treatment with tetradecanoylphobol-13-acetate, LH or forskolin could still induce NGFI-B expression, and RO 31-8220 or the PKCzeta pseudosubstrate peptide inhibited LH- or forskolin-induced NGFI-B expression. Furthermore, overexpression of dominant-negative PKCzeta in primary granulosa cells using a replication-defective adenovirus vector resulted in the suppression of LH- or forskolin-induced NGFI-B expression. Our findings demonstrate that PKCzeta, which is activated by LH or forskolin, contributes to the induction of NGFI-B in granulosa cells of preovulatory follicles. PMID- 17416459 TI - Study of the separation efficiency of a demister vane with response surface methodology. AB - Numerical simulations of a demister vane with various geometries and operating conditions were performed to study the separation efficiency. The numerical solutions were carried out using commercial CFD code Fluent 6.1. A prediction model of the separation efficiency was obtained based on response surface methodology by means of the statistical software program Minitab V14. The results show that not only the vane spacing and flue gas velocity, but also vane height (including height of curve and upright region) and vane turning angles, play an important role in influencing the separation efficiency. Compared with some experimental and simulative conclusions, the results indicate that present prediction model can estimate the effects of different geometries and operation conditions on the separation efficiency, and can direct the optimum design of demisters. PMID- 17416460 TI - Investigation of the catalytic wet peroxide oxidation of phenol over different types of Cu/ZSM-5 catalyst. AB - In this work oxidation of phenol with hydrogen peroxide on Cu/ZSM-5 catalysts was studied. The catalysts samples were prepared by two different methods: by ionic exchange from the protonic form of commercial ZSM-5 zeolite, and by direct hydrothermal synthesis. Characterization of the catalysts extends to X-ray diffraction (XRD), while the adsorption techniques were used for the measurement of the specific surface area. The catalytic tests were carried out in a stainless steel Parr reactor in batch operation mode at the atmospheric pressure and the temperature range from 50 to 80 degrees C. The mass ratio of the active metal component on the zeolite was in the range of 1.62-3.24 wt.%. for catalyst prepared by direct hydrothermal synthesis and 2.23-3.52 wt.% for catalyst prepared by ion exchange method. The initial concentration of phenol and hydrogen peroxide was 0.01 and 0.1 mol dm(-3), respectively. The influence of different methods of Cu/ZSM-5 preparation on their catalytic performance was monitored in terms of phenol conversion and degree of metal leached into aqueous solution. PMID- 17416461 TI - Copper and cadmium adsorption on pellets made from fired coal fly ash. AB - Studies on the utilization of low cost adsorbents for removal of heavy metals from wastewaters are gaining attention. Fired coal fly ash, a solid by-product that is produced in power plants worldwide in million of tonnes, has attracted researchers' interest. In this work, fly ash was shaped into pellets that have diameter in-between 3-8mm, high relative porosity and very good mechanical strength. The pellets were used in adsorption experiments for the removal of copper and cadmium ions from aqueous solutions. The effect of agitation rate, equilibration time, pH of solution and initial metal concentration were studied. The adsorption of both cations follows pseudo-second order kinetics reaching equilibrium after an equilibration time of 72 h. The experimental results for copper and cadmium adsorption fit well to a Langmuirian type isotherm. The calculated adsorption capacities of pellets for copper and cadmium were 20.92 and 18.98 mg/g, respectively. Desorption experiments were performed in several extraction media. The results showed that both metals were desorbed substantially from pellets under acidic solutions. For this reason, metal saturated pellets were encapsulated in concrete blocks synthesized from cement and raw pulverized fly ash in order to avoid metal desorption. The heavy metals immobilization after encapsulation in concrete blocks was tested through desorption tests in several aqueous media. The results showed that after 2 months in acidic media with pH 2.88 and 4.98 neither copper nor cadmium were desorbed thus indicating excellent stabilization of heavy metals in the concrete matrix. As a conclusion, the results showed that fly ash shaped into pellets could be considered as a potential adsorbent for the removal of copper and cadmium from wastewaters. Moreover, the paper proposes an efficient and simple stabilization process of the utilized adsorbents thus guarantying their safe disposal in industrial landfills and eliminating the risk of pollution for groundwater and other natural water receivers. PMID- 17416462 TI - Characterisation of products of tricalcium silicate hydration in the presence of heavy metals. AB - The hydration of tricalcium silicate (C(3)S) in the presence of heavy metal is very important to cement-based solidification/stabilisation (s/s) of waste. In this work, tricalcium silicate pastes and aqueous suspensions doped with nitrate salts of Zn(2+), Pb(2+), Cu(2+) and Cr(3+) were examined at different ages by X ray powder diffraction (XRD), thermal analysis (DTA/TG) and (29)Si solid-state magic angle spinning/nuclear magnetic resonance (MAS/NMR). It was found that heavy metal doping accelerated C(3)S hydration, even though Zn(2+) doping exhibited a severe retardation effect at an early period of time of C(3)S hydration. Heavy metals retarded the precipitation of portlandite due to the reduction of pH resulted from the hydrolysis of heavy metal ions during C(3)S hydration. The contents of portlandite in the control, Cr(3+)-doped, Cu(2+) doped, Pb(2+)-doped and Zn(2+)-doped C(3)S pastes aged 28 days were 16.7, 5.5, 5.5, 5.5, and <0.7%, respectively. Heavy metals co-precipitated with calcium as double hydroxides such as (Ca(2)Cr(OH)(7).3H(2)O, Ca(2)(OH)(4)4Cu(OH)(2).2H(2)O and CaZn(2)(OH)(6).2H(2)O). These compounds were identified as crystalline phases in heavy metal doping C(3)S suspensions and amorphous phases in heavy metal doping C(3)S pastes. (29)Si NMR data confirmed that heavy metals promoted the polymerisation of C-S-H gel in 1-year-old of C(3)S pastes. The average numbers of Si in C-S-H gel for the Zn(2+)-doped, Cu(2+)-doped, Cr(3+)-doped, control, and Pb(2+)-doped C(3)S pastes were 5.86, 5.11, 3.66, 3.62, and 3.52. And the corresponding Ca/Si ratios were 1.36, 1.41, 1.56, 1.57 and 1.56, respectively. This study also revealed that the presence of heavy metal facilitated the formation of calcium carbonate during C(3)S hydration process in the presence of carbon dioxide. PMID- 17416463 TI - Biological nitrate removal from wastewater of a metal-finishing industry. AB - An upflow packed bed reactor at laboratory scale has been operated for a continuous period of 5 months to investigate the technical feasibility of biological nitrate removal applied to the effluent of the coagulation sedimentation wastewater of a metal-finishing industry. The reactor was fed with industrial wastewater in a five-fold dilution to reproduce the global spill in the factory (20/80, industrial wastewater/domestic wastewater) with a concentration of nitrate between 141 and 210 gNO(3)-N/m(3). Methanol was added as a carbon source for denitrification. Inlet flow rate was progressively increased from 9 to 40 L/day (nitrogen input load from 45 to 250 gNO(3)-N/(m(3)h)). The highest observed denitrification rate was 135 gNO(3)-N/(m(3)h) at a nitrate load of 250 gNO(3)-N/(m(3)h), and removal efficiencies higher than 90% were obtained for loads up to 100 gNO(3)-N/(m(3)h). A mass relation between COD consumed and NO(3)-N removed around 3.31 was observed. Better results were achieved in a previous stage using tap water with nitrate added as a sole pollutant as a synthetic feed (critical load of 130 gNO(3)-N/(m(3)h) and denitrification rate of 200 gNO(3)-N/(m(3)h) at a nitrate load of 250 gNO(3)-N/(m(3)h)). This fact could indicate that the chemical composition of the industrial source hinders to some extent the performance of the biological process. Whatever case, results demonstrated the viability of the denitrification process for the global industrial wastewater. A simple model based on Monod kinetics for substrate consumption, and constant biomass concentration was applied to model the industrial wastewater treatment, and a reasonably good fitting was obtained. PMID- 17416464 TI - Distribution of heavy metals in Lakes Doirani and Kerkini, Northern Greece. AB - The distribution of heavy metals in two lakes of high ecological significance, Doirani and Kerkini, located in Northern Greece was studied. Eight metals (Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) were determined in water, total suspended solids, fine and coarse sediments. Moreover, the modified BCR fractionation scheme was employed in sediments and suspended solids to determine soluble, oxidisable, reducible and residual fractions of metals. The Lake Doirani presents higher metal concentrations in aqueous phase than Lake Kerkini; Cd, Cu, Ni, Pb and Zn are above the chronic freshwater quality criteria for aquatic life. In both lakes, Fe and Mn are the most abundant elements in total suspended solids whereas Cd the less abundant. The Lake Kerkini exhibits higher concentrations of all the examined metals in sediments comparing to the Lake Doirani, however the concentrations are lower than the sediment quality guidelines. Cd in sediments is mainly in soluble fraction, Pb and Cu exhibit significant oxidisable fractions whereas, Cr and Fe associated mainly with residual fraction. PMID- 17416465 TI - Studies on sorption of Cd(II) on Tata chromite mine overburden. AB - Chromite mine overburden containing iron as oxide/hydroxide, a waste material generated in chromite mines was used as sorbent for cadmium. The iron content of material was 43.75% with a specific surface area of 50.8m(2)/g. Batch experiments were conducted to study the sorption behavior of Cd(II) on this material. The variable experimental parameters were: time, pH, temperature, Cd(II) and sample concentration. The point of zero charge (PZC) of the overburden sample was experimentally determined as 6.48 which shifted to a pH of 7.8 when the sample was equilibrated with 100 mg/L Cd(II) solution. Maximum loading capacity of the overburden sample was found to be approximately 19 mg Cd/g of material. It was observed that within 30 min the sorption attains equilibrium. Hence, the sorption data generated at 30 min with various initial Cd(II) concentrations and temperatures were taken to evaluate the thermodynamic parameters, i.e., DeltaG degrees , DeltaH degrees and DeltaS degrees . The DeltaG degrees values reflect the feasibility of the metal removal from aqueous solution. The negative values of DeltaH degrees confirmed the exothermic sorption of cadmium and the positive DeltaS degrees values suggested the increased randomness at the solid-solution interface. The sorption data fitted well to both the Langmuir and Freundlich isotherm models indicating a monolayer sorption. The value of Freundlich parameter 'n' (n is indicative of sorption intensity) lying between 1.46 and 1.59 shows that the surface of the sorbent is heterogeneous in nature. PMID- 17416466 TI - Genetic characterisation of six Sarcocystis species from reindeer (Rangifer tarandus tarandus) in Norway based on the small subunit rRNA gene. AB - Six Sarcocystis species, i.e. Sarcocystis grueneri, Sarcocystis rangi, Sarcocystis tarandivulpes, Sarcocystis hardangeri, Sarcocystis rangiferi and Sarcocystis tarandi have previously been described from reindeer based on sarcocyst morphology. In order to validate and expand the species descriptions, the complete small subunit (ssu) rRNA gene was sequenced and used to genetically characterise the six species. The aim was to reveal possible genetic variation in the ssu rRNA gene within each Sarcocystis species, between the species from reindeer, but also between the reindeer species and related Sarcocystis species characterised in other studies. Muscle tissue from the heart and diaphragm was sampled from 18 adult semi-domesticated reindeer at a field abattoir in northern Norway. Sarcocysts were excised from the tissue and classified into one of the six known Sarcocystis species based on their morphology. Two cysts of each of the six species from two different animals were randomly selected for further DNA analyses. The complete ssu rRNA gene was amplified by the polymerase chain reaction (PCR) and subsequently sequenced. The complete ssu rRNA gene sequences were used to compare the six species with each other and with other previously sequenced Sarcocystis species retrieved from GenBank. There was little sequence variation between two isolates of the same species, but the six species differed from each other by insertions, deletions and single nucleotide polymorphisms (SNPs), mainly located in variable regions of the gene. The identity between the six species from reindeer was approximately the same as when other Sarcocystis species using a different intermediate host were compared with each other. This study supported previous findings of reindeer being the intermediate host for at least six Sarcocystis species and the results also indicated the existence of certain nucleotide positions within the ssu rRNA gene that are unique to Sarcocystis species with a canine definitive host. PMID- 17416467 TI - A longitudinal study of seroprevalence and seroconversion of Neospora caninum infection in dairy cattle in northeast Thailand. AB - A long-term study was carried out in 11 dairy herds in the Khon Kaen province of northeast Thailand between August 2001 and November 2004. The objective was to investigate seroprevalence dynamics of Neospora caninum infection in the herds and to demonstrate patterns of seroconversion in individual cattle. Each herd was visited once a year, in total four times, and sera from cattle > 3 months of age and farm dogs as well as a sample from the bulk milk were collected. All samples were analysed for presence of specific antibodies by an N. caninum iscom ELISA. The overall percentage of antibody-positive cattle was constant and varied only between 10 and 13% over the 4 years, but the variation in within-herd seroprevalence between herds was substantial. Two herds had > or = 20% seropositive animals at all samplings and consistently high bulk milk OD, whereas two herds had no seropositive animal at the last two samplings and low bulk milk OD. Five herds had a decreasing trend of within-herd seroprevalence, whereas the remaining six herds had a higher portion of test-positive individuals at the end of the study. A total of 424 individuals were sampled more than once; 344 (81%) and 32 (8%) were consistently antibody-negative and antibody-positive, respectively. The proportions of animals that changed from being seronegative to seropositive and from being seropositive to seronegative between the years were 3.9-4.6% and 19-39%, respectively. Apparent vertical and horizontal transmission rates were 58% (95% CI; 44-71%) and 5% (95% CI; 3-7%), respectively. In conclusion, the overall percentage of N. caninum antibody-positive cattle was constant over the years, but the within-herd seroprevalence varied substantially between the herds. Seroconversions were likely to occur in individual cattle although most animals had consistent serological status throughout the study. PMID- 17416468 TI - A simple prognostic scoring system for patients with unresectable hepatocellular carcinoma treated by chemo-embolization. AB - BACKGROUND: Hepatocellular carcinoma (HCC) comprises heterogeneous groups of patients with differing outcomes. METHODS: In order to attempt to identify patient sub-sets, we retrospectively examined the records of 750 patients with biopsy-proven unresectable HCC, who were treated with hepatic artery chemo embolization and were followed till death. We used the Cox proportional hazard model, as it was neutral with respect to prejudged cut-off between short and long survival. RESULTS: On univariate analysis, we found a shorter survival to be associated with male gender, presence of cirrhosis, portal hypertension, portal vein thrombosis and elevations of bilirubin, GGTP, ALKP, AFP, DCP, PT and albumin. Five factors were found to be statistically significant (p<0.05) on multivariate analysis, namely presence of cirrhosis or ascites and elevations of AFP, ALKP or GGTP. We developed a simplified scoring system based upon the sum of the hazard ratios of each of these five factors. By combining the two factors with the heaviest hazard ratios from our multivariable analysis, namely AFP (+ = >100 ng/mL) and ALKP (+ = >100 IU/mL), we found a simple parsimonious prognostic tool, which segregated the patients into survival groups, namely AFP- ALKP-; either AFP+ or ALKP+; and AFP+ ALKP+; these three groups corresponded to a 24 month survival of 70%, 32% and 12%, respectively. CONCLUSION: we found that only two lab functions, AFP and ALKP levels, in our large HCC patient cohort undergoing hepatic artery chemo-embolization, had prognostic significance. PMID- 17416469 TI - Rapid acquisition of preference in concurrent schedules: effects of reinforcement amount. AB - Four pigeons were trained on concurrent variable-interval 30-s schedules. Relative reinforcer amounts arranged across the two alternatives were varied across sessions according to a pseudorandom binary sequence [cf., Hunter, I., Davison, M., 1985. Determination of a behavioral transfer function: white-noise analysis of session-to-session response-ratio dynamics on concurrent VI schedules. J. Exp. Anal. Behav. 43, 43-59]; the ratios (left/right) were either 1/7 or 7/1. Reinforcer amount was manipulated by varying the number of 1.2s hopper presentations. Sessions ended after 30 reinforcers (15 for each alternative). After approximately 30 sessions, response ratios for all pigeons began to track the changes in amount ratio (i.e., subjects' responding showed a moderate increase in sensitivity of responding to reinforcer amount). Characteristics of responding were similar to procedures in which reinforcer rate and immediacy have been manipulated, although sensitivity estimates for amount were lower than those previously obtained with rate and immediacy. This procedure may serve as a useful method for studying the effects of certain environmental manipulations (e.g., drug administration) on sensitivity to reinforcer amount. PMID- 17416470 TI - Evaluation of OPRM1 variants in heroin dependence by family-based association testing and meta-analysis. AB - OPRM1, which codes for the mu-opioid receptor, is the most frequently studied candidate gene for opioid dependence. Despite numerous allelic association studies, no definitive conclusion has been reached regarding the role of OPRM1 polymorphisms in determining risk for opioid dependence. We attempted to resolve this by conducting a family-based association study and meta-analysis which may be more robust and powerful, respectively, than traditional case-control analyses. First, we genotyped three single nucleotide polymorphisms (SNPs) of OPRM1 in 1208 individuals from 473 Han Chinese families ascertained on the basis of having two or more siblings with DSM-IV-defined opioid dependence. The Val6Ala and Arg111His SNPs were detected, but with low minor allele frequencies (0.002 and 0.001, respectively). The Asn40Asp SNP was more informative (minor allele frequency: 0.419), but no significant evidence was observed for either a dominant (p=0.810) or additive (p=0.406) effect of this polymorphism on risk for opioid dependence. In addition, a meta-analysis of case-control studies of opioid dependence was performed, and found a similar lack of evidence for an association with the Asn40Asp SNP (p=0.859). Although a role of OPRM1 polymorphisms in determining risk for opioid dependence cannot be entirely discounted, a major contribution of the Asn40Asp polymorphism seems unlikely. Further analysis is warranted in samples from specific ancestral groups. In addition, it is critical that other OPRM1 variants, including all haplotype-tagging and amino-acid-coding SNPs, be tested for an influence on risk for opioid dependence, since the Asn40Asp polymorphism is only one of several hundred known mutations in the gene. PMID- 17416471 TI - Prevalence of group C rotaviruses in weaning and post-weaning pigs with enteritis. AB - Diarrheic fecal specimens collected from porcine herds were screened for the presence of group C rotaviruses using a reverse transcription-polymerase chain reaction (RT-PCR) assay. A total of 188 samples were tested and 54 were positive. When compiled these data with diagnostic results on group A rotaviruses and enteric caliciviruses we found that all but 5 group C rotavirus positive samples contained at least one additional virus. A subset of samples were subjected to nucleotide sequencing. The selected strains showed an unexpectedly wide range of nucleotide sequence heterogeneity (88.6-100%) to each other and to the reference porcine group C rotavirus strain, Cowden. The nucleotide sequence identity to the genuine bovine and human strains were, respectively, 86.8 and 87.2% or less. In conclusion, our study demonstrates that infection with group C rotavirus is frequent in Italian piggeries. The considerable rate of multiple infections requires further studies to investigate the pathogenic potential of group C rotaviruses in pigs, alone or in mixed infection, and raises challenges in the laboratory diagnosis of porcine enteric infections. PMID- 17416472 TI - Enhanced protection of modified human acidic fibroblast growth factor with polyethylene glycol against ischemia/reperfusion-induced retinal damage in rats. AB - Molecular modification with polyethylene glycol (PEGylation) is an effective approach to improve protein biostability and decrease protein immunogenic activity. To create a PEGylated recombinant human acid fibroblast growth factor (rhaFGF) and improve its bio-stability, we have produced a rhaFGF mutant (rhaFGF(ser98,132)) by replacing the 98th and the 132nd cysteine residues with serine residues. The rhaFGF(ser98,132) that retains the bioactivity of rhaFGF was then site-specifically conjugated with PEG-maleimide at the 31st cysteine residue. PEGylated rhaFGF(ser98,132) has less effect than the native rhaFGF(ser98,132) on stimulating 3T3 cell proliferation in vitro; however, its biostability at a prolonged incubation under various temperatures and resistance to trypsinization were significantly enhanced, and half-life time in vivo was elongated while its immunogenicity was significantly decreased. The physiological function of PEGylated rhaFGF(ser98,132) was evaluated in a rat model of retinal ischemia/reperfusion injury, showing that in vivo supplementation of PEGylated rhaFGF(ser98,132) provided a significantly better protection than the native rhaFGF(ser98,132) against ischemia/reperfusion-induced retinal morphological changes and lipid peroxidation. The protection is probably mediated by antioxidant protective mechanisms. PMID- 17416473 TI - Detection of lot-to-lot variations in the amorphous microstructure of lyophilized protein formulations. AB - Reconstitution of lyophilized protein formulations sometimes results in a cloudy solution, depending on the compositions and manufacturing conditions, which causes quality concerns. In this study, the lyophilized protein formulation of recombinant human Interleukin-11 (rhIL-11) was investigated using different lots with varying dissolution behaviors upon reconstitution due to differing processing conditions. In an attempt to distinguish the solid structures in the different lots, relatively new techniques such as inverse gas chromatography (IGC) and thermally stimulated depolarized current (TSDC) as well as powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) were adopted for analysis. PXRD, DSC, and IGC all failed to distinguish between the solid structures, but TSDC was able to discern the differences. Interestingly, TSDC suggested that the variations in dissolution behavior were attributable to the differences in molecular mobility and the micro heterogeneity of amorphous components in the solid structures. Since even the cloudiest reconstituted solutions became transparent in several minutes, it was likely that the differences in the solid structures of the different lots of lyophilized cakes were slight. This study demonstrates the usefulness of TSDC in the analysis of lot-to-lot variations in amorphous pharmaceuticals. PMID- 17416474 TI - Cochlear implants and ex vivo BDNF gene therapy protect spiral ganglion neurons. AB - Spiral ganglion neurons often degenerate in the deaf ear, compromising the function of cochlear implants. Cochlear implant function can be improved by good preservation of the spiral ganglion neurons, which are the target of electrical stimulation by the implant. Brain derived neurotrophic factor (BDNF) has previously been shown to enhance spiral ganglion survival in experimentally deafened ears. Providing enhanced levels of BDNF in human ears may be accomplished by one of several different methods. The goal of these experiments was to test a modified design of the cochlear implant electrode that includes a coating of fibroblast cells transduced by a viral vector with a BDNF gene insert. To accomplish this type of ex vivo gene transfer, we transduced guinea pig fibroblasts with an adenovirus with a BDNF gene cassette insert, and determined that these cells secreted BDNF. We then attached BDNF-secreting cells to the cochlear implant electrode via an agarose gel, and implanted the electrode in the scala tympani. We determined that the BDNF expressing electrodes were able to preserve significantly more spiral ganglion neurons in the basal turns of the cochlea after 48 days of implantation when compared to control electrodes. This protective effect decreased in the higher cochlear turns. The data demonstrate the feasibility of combining cochlear implant therapy with ex vivo gene transfer for enhancing spiral ganglion neuron survival. PMID- 17416476 TI - Frontotemporal abnormal gyration with infantile spasms in identical twins. AB - We describe the case of 10-year-old twin girls, who were born to nonconsanguineous parents and showed pes equinovarus, infantile spasms, and severe psychomotor retardation. Neither patient showed marked microcephaly. Neuroimaging revealed a simplified gyral pattern in the bilateral frontotemporal areas, a reduced volume of cerebral white matter, and a thin corpus callosum in both patients. The cerebral cortex presented with significant thickness in the anterior temporal region at the age of 10 years, which suggest that the simplified gyri can evolve into the pachygyria during postnatal period. These cases may represent a hereditary brain malformation with a distinct pattern of abnormal cortical gyration. The age-dependent change in appearance of dysgenetic cortex should be further explored in cases with brain malformations. PMID- 17416477 TI - REMOVED: Assessment of Pulmonary Venous Variation by Multidetector row CT: Clinical Implication for Catheter Ablation Techniques for Atrial Fibrillation. AB - This article has been removed, consistent with Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). The Publisher apologizes for any inconvenience this may cause. PMID- 17416478 TI - Effects of maternal hyperhomocysteinemia induced by high methionine diet on the learning and memory performance in offspring. AB - In this study, we suggest that chronic maternal hyperhomocysteinemia results in learning deficits in the offspring due to delayed brain maturation and altered expression pattern of neural cell adhesion molecule. Although the deleterious effects of hyperhomocysteinemia were extensively investigated in the adults, there is no clear evidence suggesting its action on the developing fetal rat brain and cognitive functions of the offspring. Therefore, in the present work we aimed to investigate effects of maternal hyperhomocysteinemia on the fetal brain development and on the behavior of the offspring. A group of pregnant rats received daily methionine (1 g/kg body weight) dissolved in drinking water to induce maternal hyperhomocysteinemia, starting in the beginning of gestational day 0. The levels of glial fibrillary acidic protein, S100B protein, and neural cell adhesion molecule were determined in the tissue samples from the pups. Learning and memory performances of the young-adult offsprings were tested using Morris water maze test. There were significant reductions in the expressions of glial fibrillary acidic protein and S100B protein in the brains of maternally hyperhomocysteinemic pups on postnatal day 1, suggesting that hyperhomocysteinemia delays brain maturation. In conclusion, maternal hyperhomocysteinemia changes the expression pattern of neural cell adhesion molecule and therefore leads to an impairment in the learning performance of the offspring. PMID- 17416479 TI - Development and process evaluation of an internet-based physical activity behaviour change program. AB - OBJECTIVES: Limited research exists addressing issues of website development and attitudes towards websites that promote physical activity. This paper describes the design and process evaluation of a physical activity intervention, from a randomised trial (RT) that examined the effectiveness of delivery modes (face-to face, Internet-mediated (combined Internet and face-to-face) and Internet-only). METHODS: Evaluation surveys were completed at the end of the RT intervention (n=159), and a random sample (n=23) participated in focus group discussions. Differences were assessed using ANOVA, t-tests, and qualitative analysis. RESULTS: Face-to-face participants were more satisfied with their group allocation than the Internet groups. Internet groups reported accessing the Internet at home, and that they liked the self-paced nature of the website. Difficulties associated with time and motivation were raised by the Internet groups. No differences were found for content acceptability and credibility. Website usability scores were high for both Internet groups. CONCLUSION: These findings support the use of the Internet in terms of content satisfaction and usability; however, issues were raised regarding the acceptability of an Internet based program when traditional face-to-face delivery options are available. PRACTICE IMPLICATIONS: Identifying avenues to support user perceptions of 'motivation', and mechanisms for enhancing engagement and retention, and user acceptability are required. PMID- 17416480 TI - Long-term outcome of 37 patients with Wegener's granulomatosis with renal involvement. AB - OBJECTIVES: This prospective study sought to assess the long-term outcome and identify prognostic factors of patients with kidney disease related to Wegener's granulomatosis (WG). METHODS: Of 50 patients with newly diagnosed WG who were enrolled between 1990 and 1993 in a trial comparing IV and oral cyclophosphamide (CYC), 37 had renal disease, either histologically proven or diagnosed based on laboratory findings. Their principal demographic, laboratory and therapeutic data, and progression to end-stage renal disease and/or death (ESRD) provided a basis for survival analysis, using Cox proportional hazards models. RESULTS: Of the 37 patients (M/F ratio, 23/14; mean age, 55.1+/-12.1 years; antineutrophil cytoplasm antibody-positivity, 97%; ear-nose-throat involvement, 75%; pulmonary involvement, 78%; IV/oral CYC, 23/14), 36 had glomerulonephritis and one had a granulomatous renal tumor; 22 (59%) had initial serum creatinine levels >150 micromol/L. During a mean follow-up of 6.4+/-4.7 years, 15 (41%) patients died and two developed ESRD (10-year dialysis-free survival: 51+/-17%). Only one of the nine patients with renal relapses was alive without ESRD at the end of the study. According to uni- and multivariate analyses, dialysis-free survival was significantly shorter for patients with initial serum creatinine >150 micromol/L (10-year dialysis-free survival, 24+/-18% versus 89+/-21%) (hazard ratios=20.2 and 21.7; P<0.005), while the initial route of CYC administration did not influence outcome. CONCLUSION: These observations confirm the poor survival and functional outcome associated with renal involvement of WG and highlight the strong prognostic impact of renal function at diagnosis and of renal relapses during follow-up. Conversely, the initial route of CYC administration appears to have no effect on survival. PMID- 17416481 TI - Effect of endoplasmic reticulum stress preconditioning on cytotoxicity of clinically relevant nephrotoxins in renal cell lines. AB - The cytoprotection of LLC-PK1 cells afforded by endoplasmic reticulum (ER) stress preconditioning suggests that the ER plays an important role during drug-induced renal toxicity. However, in vitro studies have been largely limited to LLC-PK1 cells and model toxins. Therefore, we tested the hypothesis that cytoprotection following ER stress preconditioning is a common property of renal cell lines (LLC PK1 (pig), NRK-52E (rat), HEK293 (human), MDCK (dog)) and extends to clinically relevant nephrotoxins. ER stress inducers (tunicamycin, thapsigargin and oxidized dithiothreitol (DTTox)) resulted in a dose-dependent increase in GRP78 and GRP94 stress protein expression, but the magnitude of induction was cell line- and inducer-dependent. Toxicity of the model toxins iodoacetamide and tert butylhydroperoxide was modified by preconditioning. DTTox was effective in decreasing the toxicity in all cell lines, but protection was variable with tunicamycin and thapsigargin. Toxicity of clinically relevant drugs (cisplatin, gentamicin, glyoxylate, cyclosporine A, p-aminophenol) was significantly decreased in cells preconditioned by tunicamycin or DTTox. These results demonstrate that ER stress preconditioning offers cytoprotection against clinically relevant nephrotoxins in renal cell lines from multiple species, although there were qualitative and quantitative differences between the cell lines. These results support the hypothesis that ER is involved in drug-induced renal toxicity. PMID- 17416482 TI - The oxidative damage of butenolide to isolated erythrocyte membranes. AB - Butenolide (CAS No. 16275-44-8), a mycotoxin produced by several Fusarium species, has been shown to be a potential risk factor for animal and human health. This study was undertaken to investigate the potential oxidative damage of butenolide to biomembranes in vitro using the erythrocyte membrane model. Following exposure of isolated rat erythrocyte membranes to butenolide, the extent of oxidative damage was assessed by measuring lipid peroxidation, -SH groups content, Ca2+/Mg2+-ATPase and Na+/K+-ATPase activities, and conformational changes in membrane proteins. It was observed that butenolide resulted in a significant lipid peroxidation, revealed by a concentration-dependent increase in the level of thiobarbituric acid reactive substances (TBARS). Similarly, this toxin induced a concentration-dependent decrease in the content of membrane total -SH groups, as well as free -SH groups. Membrane-bound enzymes were also impaired by the toxin, demonstrated by the marked inhibition of the activities of Na+/K+ ATPase and Ca2+/Mg2+-ATPase. Conformational changes in membrane proteins were determined using electron paramagnetic resonance (EPR) spin labeling. Butenolide caused an increase in the ratio of weakly to strongly immobilized components (W/S ratio) in a manner of concentration-dependent, indicating conformational changes in membrane proteins occurred. In conclusion, these findings indicate that butenolide is capable of inducing significant oxidative damage to membrane lipids and proteins. PMID- 17416483 TI - Arsenite-induced apoptosis is prevented by antioxidants in zebrafish liver cell line. AB - This study evaluated oxidative stress-induced apoptosis as a possible mechanism of arsenite toxicity in zebrafish liver cell line (ZFL cells). The heat shock protein 70 (HSP70), a chaperone protein, appears to provide protection against oxidative stress and apoptosis. Using the MTT assay, we demonstrated that survival of ZFL cells treated with arsenite for 24h decreased in a dose-dependent manner. The possible mechanisms that promote the cytotoxicity of arsenite were addressed. Cell viability assays revealed that arsenite caused a dose-dependent increase in cell death, and pretreatment of the ZFL cells with antioxidants blunted these effects. Antioxidants such as N-acetyl-cysteine (NAC, 5 mM) and dithiothreitol (DTT, 80 microM) significantly prevented ZFL cells from arsenite induced death. Nuclear staining was performed using 1 microg/ml Hoechst, and cells were analyzed with a fluorescent microscope. Arsenite (30 microM) induced massive apoptosis that was identified by morphology and condensation and fragmentation of the nuclei of the ZFL cells. Pretreatment with NAC or DTT before arsenite insult effectively protected the cells against oxidative stress-induced apoptosis from the arsenite. Using a transfected human hsp 70 promoter-enhanced green fluorescent protein (EGFP) reporter, pHhsp70-EGFP, the induction of HSP70 against oxidative stress-induced apoptosis by arsenite was observed. The induction of HSP70 by arsenite increased in a dose-dependent manner, and pretreatment of transfected ZFL cells with NAC or DTT before arsenite insult reduced EGFP expression. Taken together, our results provide evidence that stimulation of the heat shock response is a sensitive biomarker of arsenic exposure and that arsenite causes oxidative stress-induced apoptosis in ZFL cells. PMID- 17416484 TI - Detecting attention-deficit/hyperactivity disorder (ADHD) in adults with intellectual disability The use of Conners' Adult ADHD Rating Scales (CAARS). AB - There is an increasing interest in the diagnosis of attention deficit/hyperactivity disorder (ADHD) in adulthood. It is also thought that ADHD is more prevalent in the field of intellectual disability (ID) than in the general population, but there are not many experimental studies. Since ADHD diagnosis in adults is more difficult, specific rating scales correlated to the main diagnostic systems have been created but have not been applied to people with ID. This work presents an application of an ADHD screening rating scale, the Conners' Adult ADHD Rating Scales (CAARS) screening version to 46 adults with ID. The resulting prevalence of "ADHD-positive" was 19.6%. These data are in accord with results reported in the general adult literature. Our data suggest that ID and attention disorders can co-occur. Therefore, not only can ADHD be a valid psychiatric diagnosis for a child with ID but for an adult with ID as well. The CAARS can be considered a useful clinical instrument to survey ADHD in ID. PMID- 17416485 TI - Concerning the differentiability of the energy function in vector quantization algorithms. AB - The adaptation rule of Vector Quantization algorithms, and consequently the convergence of the generated sequence, depends on the existence and properties of a function called the energy function, defined on a topological manifold. Our aim is to investigate the conditions of existence of such a function for a class of algorithms including the well-known 'K-means' and 'Self-Organizing Map' algorithms. The results presented here extend several previous studies and show that the energy function is not always a potential but at least the uniform limit of a series of potential functions which we call a pseudo-potential. It also shows that a large number of existing vector quantization algorithms developed by the Artificial Neural Networks community fall into this class. The framework we define opens the way to studying the convergence of all the corresponding adaptation rules at once, and a theorem gives promising insights in that direction. PMID- 17416486 TI - Evidence against enhanced glutamate transport in the anticonvulsant mechanism of the ketogenic diet. AB - Excessive glutamatergic neurotransmission is considered an underlying factor of epilepsy. Energy-dependent glutamate transporters clear extracellular glutamate to limit neuronal excitability. Evidence suggests that reduced expression and/or activity of glutamate transporters contribute to hyperexcitability and progressive seizure activity in rats. By comparison, treatment with the anticonvulsant ketogenic diet (KD) results in increased mRNA expression of the neuronal glutamate transporter EAAC1, elevated energy reserves, and an increased resistance to seizures in rats. The goal of the current study was to determine whether the expression and/or re-uptake activity of glutamate transporters were elevated in hippocampal tissue of rats after KD treatment. Rats were fed either a ketogenic- or control diet for 4-5 weeks. Western blot analysis showed that protein levels of EAAC1, GLT-1 and GLAST glutamate transporters were not changed in hippocampus, cerebral cortex, or cerebellum after KD. Electron microscopic evidence indicated that the KD did not affect hippocampal EAAC1 distribution. In addition, the re-uptake activity of (3)H-glutamate into hippocampal proteoliposomes was similar in both KD and control tissue extracts. These multiple studies suggest that the anticonvulsant nature of the KD does not stem from enhanced glutamate re-uptake. PMID- 17416487 TI - FK506 inhibits the enhancing effects of TGF-beta on wound healing in a rabbit dermal ulcer model. PMID- 17416488 TI - Insular cortex and neuropsychiatric disorders: a review of recent literature. AB - The insular cortex is located in the centre of the cerebral hemisphere, having connections with the primary and secondary somatosensory areas, anterior cingulate cortex, amygdaloid body, prefrontal cortex, superior temporal gyrus, temporal pole, orbitofrontal cortex, frontal and parietal opercula, primary and association auditory cortices, visual association cortex, olfactory bulb, hippocampus, entorhinal cortex, and motor cortex. Accordingly, dense connections exist among insular cortex neurons. The insular cortex is involved in the processing of visceral sensory, visceral motor, vestibular, attention, pain, emotion, verbal, motor information, inputs related to music and eating, in addition to gustatory, olfactory, visual, auditory, and tactile data. In this article, the literature on the relationship between the insular cortex and neuropsychiatric disorders was summarized following a computer search of the Pub Med database. Recent neuroimaging data, including voxel based morphometry, PET and fMRI, revealed that the insular cortex was involved in various neuropsychiatric diseases such as mood disorders, panic disorders, PTSD, obsessive-compulsive disorders, eating disorders, and schizophrenia. Investigations of functions and connections of the insular cortex suggest that sensory information including gustatory, olfactory, visual, auditory, and tactile inputs converge on the insular cortex, and that these multimodal sensory information may be integrated there. PMID- 17416489 TI - Permeation of permanently positive charged molecules through artificial membranes -influence of physico-chemical properties. AB - The aim of this study was to investigate the permeation properties of 20 permanently positive charged molecules in the parallel artificial membrane permeability assay (PAMPA). Eight of them were derivatives of the N-alkyl isoquinolinium salt and 12 were congeners of the dye rhodamine 110. Five out of 12 molecules from the rhodamine 110 series have one additional carboxylic group and two have two carboxylic acids. The experimentally derived effective permeability values (P(e)) cover a range of 3-4 log units. Ten compounds showed low permeabilities (P(e)<0.1x10(-6)cm/s), four medium permeabilities (0.1x10(-6)< or =P(e)<1x10(-6)cm/s) and six were highly permeable (P(e)> or =1x10(-6)cm/s). In addition, computational models were built with a number of calculated molecular descriptors and evaluated for their ability to predict membrane permeability. It turned out that the experimental P(e) values can be explained by electronic properties and parameters describing the shape of molecules. This work provides evidence that permanently charged molecules can have high passive membrane permeabilities provided that the charge can be spread over several aromatic ring systems. PMID- 17416490 TI - [Rotavirus vaccines and the risk of intussusception]. PMID- 17416491 TI - [Epilepsy surgery in children]. PMID- 17416492 TI - [A project of the French National Birth Cohort: ELFE]. PMID- 17416493 TI - [Why does asthma remain an undertreated disease? How about patient's responsibility?]. PMID- 17416494 TI - [Current concepts on risk and protective factors for sudden infant death syndrome]. PMID- 17416495 TI - [Who are exactly giften children?]. PMID- 17416496 TI - [Children in foster and residential care, 20 years later]. PMID- 17416497 TI - [Use of rapid antigen detection tests in unusual infections by Group A beta hemolytic streptococcus]. PMID- 17416498 TI - [Unethical situations in international adoption]. PMID- 17416499 TI - [Angiomas: therapeutic indications]. PMID- 17416500 TI - [Pediatric vascular malformations: embolization]. PMID- 17416501 TI - [Sudden infant death: from research tracts to the observed reality]. PMID- 17416502 TI - [Transition in adult care for HIV perinatally infected adolescents and young adults]. PMID- 17416503 TI - [Food allergy: during childhood or all the life?]. PMID- 17416504 TI - [Sedation and analgesia in children: regional anaesthesia]. PMID- 17416505 TI - [Fertility disorders in endocrinopathies]. PMID- 17416506 TI - [Surveillance of sudden unexpected deaths in infants less than 2 years]. PMID- 17416507 TI - [Medical management of a newborn with a congenital abnormality of the kidney and the urinary tract (CAKUT)]. PMID- 17416508 TI - Effects of rhamnolipid-biosurfactant on cell surface of Pseudomonas aeruginosa. AB - The effect of rhamnolipid-biosurfactant produced by Pseudomonas sp. PS-17 on cell surface structures of Pseudomonas aeruginosa NBIMCC 1390 was studied. The results demonstrated that the rhamnolipid at concentrations below and above CMC provoked a multi-component response of the bacterial cells without affecting their growth and viability. Above CMC, the rhamnolipid caused reduction of total cellular LPS content of 22%, which can be associated with an increase in cell hydrophobicity to 31% adherence. The rhamnolipid-biosurfactant at concentration below CMC did not affect the LPS component of the bacterial outer membrane but caused changes in OMP composition of P. aeruginosa. Examination of the OMP profiles revealed that the amount of major proteins (Opr F, Opr D, Opr J and Opr M) markedly decreased. To our knowledge this is the first report on the rhamnolipid biosurfactant interactions with bacterial cells showing changes in outer membrane proteins of P. aeruginosa. In both concentrations, the biosurfactant caused changes in cell surface morphology. The results indicate that the rhamnolipid biosurfactant from Pseudomonas sp. PS-17 has a potential application in the relatively new field of biomedicine. PMID- 17416509 TI - Assessment and comparison of the environmental performances of a regional incinerator network. AB - In Emilia-Romagna region (Northern Italy) the integrated waste treatment system consists of material collection and recycling, incineration with energy recovery and landfill as final disposal. In particular, at least one incineration plant is working in almost every province of the region. In this work, a screening life cycle assessment approach is applied to seven different incinerators, to compare the different plant technologies and identify the most relevant environmental impacts and processes. The characterization method used in the life cycle impact assessment step is Eco-indicator 99. The functional unit is 1 ton of waste input. As a first result, it can be noted that while the combustion systems are rather similar, the main variables are ascribable to gas cleaning options and efficiency in energy recovery, which result in quite different environmental performances. Among heavy metals, particular attention must be paid to Cd and As, due to their high toxicity, despite their low quantities. The impact due to dioxin emission is orders of magnitude lower than other contaminants (e.g., heavy metals). Furthermore, a catalytic system could be useful for a complete removal of organic contaminants and for a more effective abatement of nitrogen oxides. Finally, the environmental impact assessment sorts the various plants according to their age, i.e., the most recent plants provide the best environmental performances for the same quantity of combusted waste. PMID- 17416510 TI - CCA-treated wood disposed in landfills and life-cycle trade-offs with waste-to energy and MSW landfill disposal. AB - Chromated copper arsenate (CCA)-treated wood is a preservative treated wood construction product that grew in use in the 1970s for both residential and industrial applications. Although some countries have banned the use of the product for some applications, others have not, and the product continues to enter the waste stream from construction, demolition and remodeling projects. CCA treated wood as a solid waste is managed in various ways throughout the world. In the US, CCA-treated wood is disposed primarily within landfills; however some of the wood is combusted in waste-to-energy (WTE) facilities. In other countries, the predominant disposal option for wood, sometimes including CCA-treated wood, is combustion for the production of energy. This paper presents an estimate of the quantity of CCA-treated wood entering the disposal stream in the US, as well as an examination of the trade-offs between landfilling and WTE combustion of CCA treated wood through a life-cycle assessment and decision support tool (MSW DST). Based upon production statistics, the estimated life span and the phaseout of CCA treated wood, recent disposal projections estimate the peak US disposal rate to occur in 2008, at 9.7 million m(3). CCA-treated wood, when disposed with construction and demolition (C&D) debris and municipal solid waste (MSW), has been found to increase arsenic and chromium concentrations in leachate. For this reason, and because MSW landfills are lined, MSW landfills have been recommended as a preferred disposal option over unlined C&D debris landfills. Between landfilling and WTE for the same mass of CCA-treated wood, WTE is more expensive (nearly twice the cost), but when operated in accordance with US Environmental Protection Agency (US EPA) regulations, it produces energy and does not emit fossil carbon emissions. If the wood is managed via WTE, less landfill area is required, which could be an influential trade-off in some countries. Although metals are concentrated in the ash in the WTE scenario, the MSW landfill scenario releases a greater amount of arsenic from leachate in a more dilute form. The WTE scenario releases more chromium from the ash on an annual basis. The WTE facility and subsequent ash disposal greatly concentrates the chromium, often oxidizing it to the more toxic and mobile Cr(VI) form. Elevated arsenic and chromium concentrations in the ash leachate may increase leachate management costs. PMID- 17416511 TI - Life cycle assessment of disposal of residues from municipal solid waste incineration: recycling of bottom ash in road construction or landfilling in Denmark evaluated in the ROAD-RES model. AB - Two disposal methods for MSWI bottom ash were assessed in a new life cycle assessment (LCA) model for road construction and disposal of residues. The two scenarios evaluated in the model were: (i) landfilling of bottom ash in a coastal landfill in Denmark and (ii) recycling of bottom ash as subbase layer in an asphalted secondary road. The LCA included resource and energy consumption, and emissions associated with upgrading of bottom ash, transport, landfilling processes, incorporation of bottom ash in road, substitution of natural gravel as road construction material and leaching of heavy metals and salts from bottom ash in road as well as in landfill. Environmental impacts associated with emissions to air, fresh surface water, marine surface water, groundwater and soil were aggregated into 12 environmental impact categories: Global Warming, Photochemical Ozone Formation, Nutrient Enrichment, Acidification, Stratospheric Ozone Depletion, Human Toxicity via air/water/soil, Ecotoxicity in water/soil, and a new impact category, Stored Ecotoxicity to water/soil that accounts for the presence of heavy metals and very persistent organic compounds that in the long term might leach. Leaching of heavy metals and salts from bottom ash was estimated from a series of laboratory leaching tests. For both scenarios, Ecotoxicity(water) was, when evaluated for the first 100 yr, the most important among the twelve impact categories involved in the assessment. Human Toxicity(soil) was also important, especially for the Road scenario. When the long-term leaching of heavy metals from bottom ash was evaluated, based on the total content of heavy metals in bottom ash, all impact categories became negligible compared to the potential Stored Ecotoxicity, which was two orders of magnitudes greater than Ecotoxicity(water). Copper was the constituent that gave the strongest contributions to the ecotoxicities. The most important resources consumed were clay as liner in landfill and the groundwater resource which was potentially spoiled due to leaching of salts from bottom ash in road. The difference in environmental impacts between landfilling and utilization of bottom ash in road was marginal when these alternatives were assessed in a life cycle perspective. PMID- 17416512 TI - Activated carbon from Ceiba pentandra hulls, an agricultural waste, as an adsorbent in the removal of lead and zinc from aqueous solutions. AB - The ability of low-cost activated carbon prepared from Ceiba pentandra hulls, an agricultural waste material, for the removal of lead and zinc from aqueous solutions has been investigated. In the batch tests experimental parameters were studied, including solution pH, contact time, adsorbent dose and initial metal ions concentration. The adsorbent exhibited good sorption potential at pH 6.0. Maximum removal of lead (99.5%) and of zinc (99.1%) with 10 g/l of sorbent was observed at 50 mg/L sorbate concentration. Removals of about 60-70% occurred in 10 min, and equilibrium was attained at around 50 min for both metals. The functional groups (CO, SO,-OH) present on the carbon surface were responsible for the adsorption of metal ions. The adsorption parameters were analysed using both the Freundlich and Langmuir models. The data are better fitted by the Freundlich isotherm as compared to Langmuir model, and the adsorption capacities for lead and zinc were 25.5 and 24.1 mg/g, respectively. Kinetics of adsorption obeyed a second order rate equation and the rate constant was found to be 2.71 x 10(-2) and 2.08 x 10(-2) g/mg/min for lead and zinc, respectively. The desorption studies were carried out using dilute HCl, and the effect of HCl concentration on desorption was studied. Maximum desorptions of 85% for lead and 78% for zinc were attained with 0.15 M HCl. PMID- 17416513 TI - Integrated microsystem for non-invasive electrophysiological measurements on Xenopus oocytes. AB - We propose a new non-invasive integrated microsystem for electrophysiological measurements on Xenopus laevis oocytes. Xenopus oocyte is a well-known expression system for various kinds of ion channels, that are potential tools in drug screening. In the traditional "Two Electrode Voltage Clamp" (TEVC) method, delicate micromanipulation is required to impale an oocyte with two microelectrodes. In our system, a non-invasive electrical access to the cytoplasm is provided by permeabilizing the cell membrane with an ionophore (e.g. nystatin). Unlike the classical patch-clamp or "macropatch" techniques, this method does not require removal of the vitelline membrane. Cell handling is significantly simplified, resulting in more robust recordings with increased throughput. Moreover, because only part of the oocyte surface is exposed to reagents, the required volume of reagent solutions could be reduced by an order of magnitude compared to the TEVC method. The fabrication process for this disposable microchip, based on poly-dimethylsiloxane (PDMS) molding and glass/PDMS bonding, is cost-efficient and simple. We tested this new microdevice by recording currents in oocytes expressing the human Epithelial Sodium Channel (hENaC) for membrane potentials between -100 and +50 mV. We recorded benzamil sensitive currents with a large signal-to-noise ratio and we also obtained a benzamil concentration-inhibition curve displaying an inhibition constant IC(50) of about 50 nM, comparable to previously published values obtained with the TEVC technique. PMID- 17416514 TI - Detection of heavy metal toxicity using cardiac cell-based biosensor. AB - Biosensors incorporating mammalian cells have a distinct advantage of responding in a manner which offers insight into the physiological effect of an analyte. To investigate the potential applications of cell-based biosensors on heavy metal toxicity detection, a novel biosensor for monitoring electrophysiological activity was developed by light-addressable potentiometric sensor (LAPS). Extracellular field potentials of spontaneously beating cardiomyocytes could be recorded by LAPS in the range of 20 microV to nearly 40 microV with frequency of 0.5-3 Hz. After exposed to different heavy metal ions (Hg(2+), Pb(2+), Cd(2+), Fe(3+), Cu(2+), Zn(2+); in concentration of 10 microM), cardiomyocytes demonstrated characteristic changes in terms of beating frequency, amplitude and duration under the different toxic effects of ions in less than 15 min. This study suggests that, with the physiological monitoring, it is possible to use the cardiac cell-based biosensor to study acute and eventually chronic toxicities induced by heavy metal ions in a long-term and no-invasive way. PMID- 17416515 TI - Direct electrochemistry of glucose oxidase and electrochemical biosensing of glucose on quantum dots/carbon nanotubes electrodes. AB - Because of their unique chemical, physical and electronic properties, Quantum dots (QDs) and carbon nanotubes (CNTs) are now extremely attractive and important nanomaterials in bioanalytical applications. In this work, CdTe QDs with the size of about 3 nm were prepared and a novel electrochemical biosensing platform of glucose based on CdTe/CNTs electrode was explored. This CdTe/CNTs electrode was prepared by first mixing CdTe QDs, CNTs, Nafion, and glucose oxidase (GOD) in appropriate amounts and then modifying this mixture on the glass carbon electrode (GC). Transmission electron microscopy (TEM) was used to observe the dispersion of CdTe QDs on carbon nanotubes and cyclic voltammetry (CV) was used to investigate the electrochemical behavior of the CdTe/CNTs electrode. A pair of well-defined quasi-reversible redox peaks of glucose oxidase were obtained at the CdTe/CNTs based enzyme electrode by direct electron transfer between the protein and the electrode. The immobilized glucose oxidase could retain bioactivity and catalyze the reduction of dissolved oxygen. Due to the synergy between the CdTe QDs and CNTs, this novel biosensing platform based on QDs/CNTs electrode responded even more sensitively than that based on GC electrode modified by CdTe QDs or CNTs alone. The inexpensive, reliable and sensitive sensing platform based on QDs/CNTs electrode provides wide potential applications in clinical, environmental, and food analysis. PMID- 17416516 TI - Optically responsive nanoparticle layers for the label-free analysis of biospecific interactions in array formats. AB - A novel nanocomposite surface is prepared by coating surface-adsorbed dielectric colloidal particles with a contiguous layer of gold nanoparticles. The resulting surface shows pronounced optical extinction in reflection with the extinction peaks located in the UV-Vis and NIR region of the electromagnetic spectrum. The peak positions of these maxima change very sensitively with the adsorption of organic molecules onto the surface. For the adsorption of a monolayer of octadecanethiol, we observe a peak shift of 55 nm on average, which is about five times that of established label-free sensing methods based on propagating and localized surface plasmons. In a first proof-of-principle experiment, the interaction of peptides with specific antibodies has been detected without labeling by means of a fiber-optical set-up with microscopic lateral resolution. To avoid crosstalk in high-density arrays, the optically responsive surface areas can be locally separated on a micro- or even nanometer scale. Accordingly, the newly developed optically responsive surfaces are well suited for integration into high-density peptide or DNA arrays as demanded in genomics, proteomics, and biomedical research in general. PMID- 17416517 TI - Blending effects of biodiesels on oxidation stability and low temperature flow properties. AB - To improve the oxidation stability and the low temperature flow properties of a biodiesel mixture, the dependence of the oxidation stability and the cold filter plugging point (CFPP) on the fatty acid compositions was examined. Three different kinds of biodiesels, palm, rapeseed, and soybean biodiesels, were blended with the different weight ratios. The oxidation stability and the CFPP of the blended biodiesels had a close relationship with the compositions of the major fatty acid components. The oxidation stability of the blended biodiesels decreased as the total contents of the linoleic and linolenic acids increased. The correlation was obtained as Y=117.9295/X+2.5905 (07000-fold). The masking of its highly hydrophilic groups led to the creation of related prodrug 28, which demonstrated an anticoagulant effect after oral dosing. PMID- 17416534 TI - Stage- and tissue-expression of genes involved in the biosynthesis and signalling of ethylene in reproductive organs of damson plum (Prunus domestica L. subsp. insititia). AB - In this work, four cDNA clones (Pd-ACS1,AJ890088; Pd-ETR1 and Pd-ERS1, AJ890092, AJ890091; and Pd-CTR1, AJ890089) encoding an ACC-synthase, two putative ethylene (ET) receptors, and a putative MAPKKK, respectively, were isolated and phylogenetically characterized in Prunus domestica L. subsp. insititia. Their expression was studied by real-time PCR during flower (closed, open and senescent) and fruit (early green, late green, maturation and ripening) development of damson plum, which is climateric. While two peaks of ET production were quantified at early green and ripening stages in whole fruits, the seed was not able to produce it during maturation and ripening stages. All studied genes were differentially expressed during flower and fruit development. In general, the level of transcripts of Pd-ACS1 was higher in fruits than in flowers. However, it was noteworthy that: (1) Pd-ACS1 expression was hardly detected in closed flowers and at low levels during early green stage; and fruit development provoked a notable differential expression in seeds, and pericarp; (2) the results of Pd-ACS1 expression during fruit development suggest a preponderant role of this gene from late green stage onward. The stamen was the only floral organ in which expression of both Pd-ETR1 and Pd-ERS1 receptor genes was not significantly altered during development; however, their expression decreased concomitantly with development of pistil (only floral organ to register a net ET production when fertilized) and during first days of ovary development (the highest ET production during all fruit development). Contrary to Pd-ERS1, the level of Pd-ETR1 mRNA was temporally quite similar in the seed. With regard Pd ETR1, even its expression was very scarce during maturation of mesocarp, was stimulated during ripening. In the epicarp, Pd-ERS1 and Pd-ETR1 were low expressed during pit hardening increasing onward and decreasing during ripening. Pd-CTR1 expression was in the seed>mesocarp>>epicarp. Spatial and temporal levels of Pd-ACS1, Pd-ETR1, Pd-ERS1 and Pd-CTR1 mRNAs described in this work demonstrate that the expression of these genes is not always constitutive and that control of its transcription may play an important role in regulating the development of reproductive organs of damson plum. PMID- 17416535 TI - Post-mediastinoscopy mortality due to drug-eluting stent thrombosis. PMID- 17416536 TI - Management options in neonates and infants with critical left ventricular outflow tract obstruction. AB - Despite being one of the most common congenital cardiac abnormalities, the treatment of critical left ventricular outflow tract obstruction (LVOTO) in the neonate and infant remains a significant challenge. Critical LVOTO in the neonate and infant includes continuous and diverse spectrum of anatomic diagnoses ranging from hypoplastic left heart complex to isolated aortic valvular stenosis with otherwise normally formed left heart structures. Depending on related anatomic features, there are multiple single ventricle and biventricular management strategies available for patients with critical LVOTO; all of which carry significant risk of death and requirement for future interventions. Because of the wide array of morphologic lesions and physiologic manifestations, application of the currently available published data to guide decision making in the individual patients can be challenging. Therefore, the aim of this review is to describe the different therapeutic strategies that can be applied to neonates presenting with critical LVOTO taking into consideration each patient's unique physiologic and anatomic findings. PMID- 17416537 TI - Surgery of saphenous vein graft aneurysm based on 64-slice computed tomography (CT) diagnostic assessment. PMID- 17416538 TI - Phylogenetic relationships of the Lake Tanganyika cichlid tribe Lamprologini: the story from mitochondrial DNA. AB - The Lamprologini are the most species-rich and diverse tribe of Lake Tanganyika cichlids, comprising around 90 described species. We reconstruct the most complete ( approximately 70 species) mtDNA phylogeny to date for this tribe, based on NADH dehydrogenase subunit 2 (ND2 approximately 1047 bp) and the non coding control region ( approximately 874 bp) and examine the degree to which mtDNA trees are good proxies for species trees. Phylogenetic relationships are assessed using Bayesian inference, maximum likelihood and maximum parsimony to determine the robustness of relationships. The resulting topologies are largely congruent and only the tree produced by an unpartitioned BI analysis is rejected using the non-parametric likelihood-based AU test. The trees are remarkably balanced, with two major clades consistently recovered in all analyses and with reasonable support. A smaller clade of deep-water species is also recovered. Overall support is good, when compared to some groups that have undergone adaptive radiation and rapid lineage formation. The much-expanded phylogeny of the group helps resolve the placement of some previously problematic taxa, such as Neolamprologus moori, highlighting the importance of greater taxonomic sampling. The results include a number of divergent placements of closely related species, and the following genera Neolamprologus, Lamprologus, Julidiochromis, Telmatochromis are not monophyletic, with alternative hypotheses consistent with traditional taxonomy providing a significantly worse fit to the data. We find several examples of divergent mtDNA taxa sequences of presumed closely related species. This could be due to incorrect taxonomy or to the failure of the mtDNA to reflect species relationships and may support the hypothesis that speciation within this group has been facilitated by introgressive hybridisation. PMID- 17416539 TI - Diagnostic agreement at the total hip using different DXA systems and the NHANES III database. AB - In 1997, the National Health and Nutrition Examination Survey III (NHANES III) total hip reference database was adopted for T-score derivation in an effort to optimize diagnostic agreement among densitometers from different manufacturers. Our study was undertaken to evaluate the effectiveness of the NHANES III standardized database at achieving agreement in diagnostic classification (normal, osteopenia, or osteoporosis) based on total hip T-scores comparing 2 different dual-energy X-ray absorptiometry (DXA) systems. This was a retrospective analysis of standard bilateral hip and lumbar spine scans done in duplicate for 60 women scanned on both a GE Lunar Prodigy and Hologic Delphi DXA system. Classification based on lumbar spine T-scores using manufacturer-specific databases was also compared as no standardized lumbar spine reference database exists. Subject's mean age was 62 yr (range: 47-83 yr). There was no statistically significant difference in diagnostic classification between DXA systems (Prodigy vs Delphi), with agreement (same women classified same way) of 92% at the left total hip. Agreement was 100% when T-scores were greater than or equal to -0.8 and less than or equal to -1.2. There was 90% agreement between DXA systems at the lumbar spine. For both hip and spine, all diagnostic discrepancies occurred when the T-scores were at or near transition thresholds between normal and osteopenia or osteopenia and osteoporosis. The difference in mean T-scores between DXA systems at left total hip was 0.11 vs 0.32 for lumbar spine (p less than 0.001). Use of the NHANES III standardized database results in good diagnostic agreement at total hip between Prodigy and Delphi. PMID- 17416540 TI - Putative role of heparan sulfate proteoglycan expression and shedding on the proliferation and survival of cells after photodynamic therapy. AB - INTRODUCTION: Photodynamic therapy is based on the selective retention of a photosensitizer by highly proliferating cells and its activation with light at the appropriate wavelength. This combination generates reactive oxygen species that ultimately kill the cells. Some cells, however, may survive photodynamic therapy and the interaction of these cells with the extracellular matrix has profound effect in tumor biology. The knowledge of photodynamic therapy action on the extracellular matrix has not been fully explored. It has been focused mainly on integrins, matrix metalloproteinases and on growth factors and immunological mediators. Other important molecules involved in the regulation of many cell processes are the glycosaminoglycans, polymers of disaccharide units, present on the cell surface and in the extracellular matrix. In most cases, the glycosaminoglycans occur as proteoglycans. AIMS: The purpose of the present investigation is to evaluate heparan sulfate proteoglycan expression and shedding, and its relation to the survival of the remaining cells, after a liposomal-AlClPc based photodynamic treatment. MATERIALS: A wild-type endothelial cell derived from rabbit aorta and its counterpart transfected with EJ-ras oncogene were used. RESULTS: Both cell lines presented augmented heparan sulfate proteoglycan syndecan-4 mRNA expression, augmented synthesis of heparan sulfate chains and increased shedding. Also, the formation of stress fibers on the border of the cells and the arrest in G(1) phase of the cell cycle was observed. CONCLUSIONS: These results show that surviving cells after photodynamic therapy exhibit changes in their morphology and cell processes that differ from that of non-treated cells, and these changes are probably hindering the cells from resuming normal proliferation. PMID- 17416541 TI - Alternative splicing in cancer: noise, functional, or systematic? AB - Pre-messenger RNA splicing is a fine-tuned process that generates multiple functional variants from individual genes. Various cell types and developmental stages regulate alternative splicing patterns differently in their generation of specific gene functions. In cancers, splicing is significantly altered, and understanding the underlying mechanisms and patterns in cancer will shed new light onto cancer biology. Cancer-specific transcript variants are promising biomarkers and targets for diagnostic, prognostic, and treatment purposes. In this review, we explore how alternative splicing cannot simply be considered as noise or an innocent bystander, but is actively regulated or deregulated in cancers. A special focus will be on aspects of cell biology and biochemistry of alternative splicing in cancer cells, addressing differences in splicing mechanisms between normal and malignant cells. The systems biology of splicing is only now applied to the field of cancer research. We explore functional annotations for some of the most intensely spliced gene classes, and provide a literature mining and clustering that reflects the most intensely investigated genes. A few well-established cancer-specific splice events, such as the CD44 antigen, are used to illustrate the potential behind the exploration of the mechanisms of their regulation. Accordingly, we describe the functional connection between the regulatory machinery (i.e., the spliceosome and its accessory proteins) and their global impact on qualitative transcript variation that are only now emerging from the use of genomic technologies such as microarrays. These studies are expected to open an entirely new level of genetic information that is currently still poorly understood. PMID- 17416542 TI - Matrix metalloproteinase-induced epithelial-mesenchymal transition: tumor progression at Snail's pace. AB - Matrix metalloproteinases (MMPs) are enzymes that digest components of the extracellular matrix (ECM) as well as cell surface receptors for soluble factors and junctional proteins involved in cell-cell and cell-ECM interactions. MMPs are involved in many physiological processes that require tissue remodeling but are also expressed in nearly all tumors, where they stimulate tumor growth, invasion, and metastasis. Previous studies have shown that expression of stromelysin-1/MMP 3 in the mammary glands of transgenic mice causes the production of invasive carcinomas; our recent investigations provide insight into how MMPs can promote the development of such tumors. We show that exposing mouse mammary epithelial cells to MMP-3 stimulates epithelial-mesenchymal transition (EMT), a phenotypic alteration in which epithelial cells acquire invasive mesenchymal characteristics. EMT is a necessary component of embryonic development, but acquisition of EMT characteristics by cancer cells facilitates invasion and metastasis. MMP-3-induced EMT involves induction of Snail, a transcription factor long studied for its role in physiological EMT but which is increasingly recognized as a factor involved in tumor progression and malignancy. In this review, we examine how MMPs and Snail function in tumor progression and how identification of an MMP-Snail signaling axis provides insight into new anticancer therapeutic strategies. PMID- 17416543 TI - Tandem gene arrays: a challenge for functional genomics. AB - In sequenced plant genomes, 15% or more of the identified genes are members of tandem-arrayed gene families. Because mutating only one gene in a duplicated pair often produces no measurable phenotype, this poses a particular challenge for functional analysis. To generate phenotypic knockouts, it is necessary to create deletions that affect multiple genes, select for rare meiotic recombination between tightly linked loci, or perform sequential mutant screens in the same plant line. Successfully implemented strategies include PCR-based screening for fast neutron-induced deletions, selection for recombination between herbicide resistance markers, and localized transposon mutagenesis. Here, we review the relative merits of current genetic approaches and discuss the prospect of site directed mutagenesis for generating elusive knockouts of tandem-arrayed gene families. PMID- 17416544 TI - Rhizosphere communication of plants, parasitic plants and AM fungi. AB - Plants use an array of secondary metabolites to defend themselves against harmful organisms and to attract others that are beneficial. However, the attraction of beneficial organisms could also lead to abuse by malevolent organisms. An exciting example of such abuse is the relationship between plants, beneficial mutualistic arbuscular mycorrhizal fungi and harmful parasitic plants. Signalling molecules called strigolactones, which are secreted by plant roots in low concentrations, induce the growth of both obligate biotrophs. Here, we review the importance of strigolactones for these two interactions and discuss possible developments that should further clarify the role of these signalling molecules in rhizosphere processes. PMID- 17416545 TI - Plant dormancy in the perennial context. AB - A key feature of the perennial life style in plants is the ability to cease meristem activity and to establish a dormant state in which the meristem is rendered insensitive to growth-promoting signals for some time before it is released and can resume growth. The seasonal cycling between growth and dormancy has received little attention despite its importance for perennial behaviour. In this review, we reconsider seasonal cycles of growth and dormancy in view of a new definition of dormancy as a state within the meristem, together with recent exciting developments in the study of perennials, particularly the identification of common signalling intermediates between flowering time and growth cessation in trees. PMID- 17416546 TI - Shopping for plastids. AB - Recent suggestions that endosymbionts in a diatom and an amoeba represent independent origins of plastids from those in plants and algae raise again the question of how many times plastids have evolved. In this Opinion article, we review the evidence for a single origin or multiple origins of primary plastids. Although the data are widely taken as supporting a single origin, we stress the assumptions underlying that view, and argue for a more cautious interpretation. We also suggest that the implicit view of plastids being acquired from single ancestors at a single point (or points) in time is an over-simplification. PMID- 17416547 TI - High-definition genome profiling for genetic marker discovery. AB - Genetic mapping is a key step towards isolating genes and genetic markers associated with phenotypic traits by elucidating their genetic positions. The success of this approach depends on precision in pinpointing genetic positions and the effectiveness of the discovery process. Recent advances in microarray technology and the increasing availability of genomic information have provided an opportunity to use microarrays to scan effectively for genetic variations at the whole-genome scale, enabling the production of high-definition gene-based genetic maps, in combination with functional analyses and identification of trait associated genetic marker candidates with high precision. In this review, we discuss the concept, process, tools and applications of microarray-based high definition genetic analysis. This post-genomics approach should help to identify causative genetic variation by uniting genetic and functional information. PMID- 17416548 TI - Vibrational spectroscopy of resveratrol. AB - In this article the authors deal with the experimental and theoretical interpretation of the vibrational spectra of trans-resveratrol (3,5,4'-trihydroxy trans-stilbene) of diverse beneficial biological activity. Infrared and Raman spectra of the compound were recorded; density functional calculations were carried out resulting in the optimized geometry and several properties of the molecule. Based on the calculated force constants, a normal coordinate analysis yielded the character of the vibrational modes and the assignment of the measured spectral bands. PMID- 17416549 TI - Pathogenomics of helicobacter. AB - The pathogenic bacterium Helicobacter pylori infects half of the human population and is one of the genetically most diverse bacterial species known. H. pylori was one of the first bacterial species whose genome was sequenced in 1997, and the first species for which two complete sequences from independent isolates were available for within-species comparisons. For almost 10 years, genomic and post genomic analysis has contributed enormously to our understanding of the pathogenesis of H. pylori infection. This review summarizes the available information, emphasizing work performed in the framework of the PathoGenoMik funding initiative (2001-2006) of the German Ministry of Education and Research. PMID- 17416550 TI - Partial aortic root remodelling for fistula between the non-coronary sinus and the right atrium. AB - This report describes a rare case of the fistula between the non-coronary sinus and the right atrium (RA) after ascending aortic replacement for chronic aortic dissection. A 67-year-old lady had been suddenly suffering from severe dyspnoea with general fatigue for a couple of days. Trans-thoracic echocardiogram in the emergency room demonstrated massive shunt flow from the non-coronary sinus to the RA with remarkable dilatation of the RA, right ventricle (RV) and inferior vena cava, similar to the rupture of sinus of Valsalva (Konno-type IV). The fistula was successfully treated by partial remodelling of the aortic root in an emergency basis because of her life-threatening illness. Some remaining diseased aortic root, which may be related to initial dissection or inappropriate use of gelatin-resorcin-formalin glue at the previous ascending aortic replacement, may cause this kind of serious events. Modified aortic root remodelling method with only diseased sinus resected was successfully applied to the localised aortic root disorder. PMID- 17416551 TI - Urgent transcatheter closure of patent foramen ovale followed by elective right sided valve surgery for decompensated carcinoid heart disease. AB - The presence of a patent foramen ovale (PFO) is associated with morbidity and mortality in patients with carcinoid heart disease (CHD). We report a 66-year-old male patient with tricuspid and pulmonary valve regurgitation secondary to CHD, who developed severe hypoxia due to a right-to-left shunt through a PFO. A 35 mm Amplatzer septal occluder was deployed to reduce the right-to-left shunt as an urgent procedure. Tricuspid and pulmonary valve replacements were electively performed using ON-X mechanical prostheses (31/33 mm and 19 mm, respectively) 70 days after the percutaneous procedure. Transcatheter closure of a PFO prior to definitive right-sided valve surgery can be a useful treatment for CHD patients with acute haemodynamic derangement. PMID- 17416552 TI - Coming of age: affiliate member profile and participation in the annual scientific meeting of the cardiac society of Australia and New Zealand. AB - BACKGROUND: Nursing, allied health and technical personnel are increasingly being recognised as pivotal in the diagnosis and management of heart disease. This recognition is mirrored in research, scholarship and professional development activities. Documenting the evolution and progression of a group's professional development is a useful strategy in informing future strategic initiatives. AIM: The purpose of this paper is to illustrate the development and participation of the Affiliates group within the Cardiac Society of Australia and New Zealand (CSANZ). METHOD: Data related to CSANZ membership, participation in the Annual Scientific Meeting as well as the number, type and ranking of abstracts were retrieved from CSANZ records for the period 1995-2003. These data were analysed using descriptive statistics. RESULTS: Since the introduction of the Affiliate member status in 1988, membership has grown steadily, as has participation of members in the governance of the CSANZ. Mean abstract grades of Affiliate members are increasingly comparable with those of the FCSANZ, Ordinary and Associate members. CONCLUSIONS: Affiliate members are increasing their profile in the highly competitive environment of the Annual Scientific Meeting, demonstrating the critical role of nursing, allied health and technical professions in cardiovascular health and science. PMID- 17416553 TI - Ex vivo gene therapy cures a blistering skin disease. AB - A recent publication that describes gene therapy treatment of a patient with an inherited blistering skin disease, epidermolysis bullosa, demonstrates for the first time that gene therapy can cure a disease of solid tissue. The treatment relies on ex vivo transduction of autologous epidermal stem cells with a normal copy of the defective gene, followed by reconstitution of the patient's skin with epithelial sheets that are grown from these genetically corrected cells. This approach holds promise for treatment not only of inherited disorders of the skin but also of other solid tissues that are becoming amenable to tissue engineering. PMID- 17416555 TI - Determination and pharmacokinetics of orientin in rabbit plasma by liquid chromatography after intravenous administration of orientin and Trollius chinensis Bunge extract. AB - A high-performance liquid chromatography (HPLC) method was developed and validated for the determination of orientin in rabbit plasma using ultraviolet (UV) absorbance detection. Orientin is the active constituent of purified herbal extract (TRO PE) from the flower of Trollius chinensis Bunge. Protein precipitation was used as the sample preparation technique. A Diamonsil C18 column (150 mmx4.6 mm, 5 microm) was equilibrated with a mobile phase composed of 0.1% acetic acid/methanol/acetonitrile (80/5/15, v/v/v). The calibration curve of orientin in rabbit plasma was linear in the concentration range of 0.530-53.0 microg/mL. This validated method was successfully applied to a pharmacokinetic study in rabbits after the intravenous administrations of orientin and TRO PE at three different doses. PMID- 17416556 TI - Halitosis associated volatiles in breath of healthy subjects. AB - BACKGROUND: Halitosis can have an intra- or extra-oral origin. In all cases, bad breath is caused by the presence of volatile organic compounds originating from the mouth or the expired air. They can be specific for certain diseases or infections. STUDY OBJECTIVE: This study explored the presence and concentration of these volatile compounds normally associated with halitosis in the breath of healthy symptomless volunteers. METHODS: Alveolar and mouth air of 40 healthy volunteers as well as environmental air were analyzed by gas chromatography-mass spectrometry (GC-MS) and by a commercially available GC device (OralChroma). RESULTS: 14 compounds, associated with halitosis could be detected. All of them except carbon disulfide, appeared to be (partly) produced endogenously and/or in the mouth. Acetone, 2-butanone, 2-pentanone and 1-propanol were common to all volunteers in both alveolar and mouth air and indole and dimethyl selenide in alveolar air. CONCLUSIONS: GC-MS seems a promising tool for differential diagnosis of halitosis, with the possibility to detect extra-oral causes, which often remain undetected unless characterized by a specific smell. PMID- 17416557 TI - Monitoring phosphatase reactions of multiple phosphorylated substrates by reversed-phase HPLC. AB - In an approach to gain insight into the sequence-dependent dephosphorylation of multiple phosphotyrosyl-containing peptides by the phosphatases SHP-1 and PTP1B, we applied a chromatographic technique for the analysis of the dephosphorylation products. Mono-, bi- and triphosphorylated reference peptides corresponding to positions 1999-2014 in the activation loop of the receptor tyrosine kinase Ros were first analyzed by reversed-phase HPLC and MALDI-TOF/TOF mass spectrometry. Then, the respective products from enzymatic treatment were investigated by HPLC and compared to the standard peptides. The results obtained in this study emphasize the advantage of monitoring phosphatase reactions for mono- and biphosphorylated peptides using the described procedure rather than spectrophotometric and fluorimetric methods that do not allow for a clear identification of the products formed. PMID- 17416554 TI - Homeostasis of exercise hyperpnea and optimal sensorimotor integration: the internal model paradigm. AB - Homeostasis is a basic tenet of biomedicine and an open problem for many physiological control systems. Among them, none has been more extensively studied and intensely debated than the dilemma of exercise hyperpnea - a paradoxical homeostatic increase of respiratory ventilation that is geared to metabolic demands instead of the normal chemoreflex mechanism. Classical control theory has led to a plethora of "feedback/feedforward control" or "set point" hypotheses for homeostatic regulation, yet so far none of them has proved satisfactory in explaining exercise hyperpnea and its interactions with other respiratory inputs. Instead, the available evidence points to a far more sophisticated respiratory controller capable of integrating multiple afferent and efferent signals in adapting the ventilatory pattern toward optimality relative to conflicting homeostatic, energetic and other objectives. This optimality principle parsimoniously mimics exercise hyperpnea, chemoreflex and a host of characteristic respiratory responses to abnormal gas exchange or mechanical loading/unloading in health and in cardiopulmonary diseases - all without resorting to a feedforward "exercise stimulus". Rather, an emergent controller signal encoding the projected metabolic level is predicted by the principle as an exercise-induced 'mental percept' or 'internal model', presumably engendered by associative learning (operant conditioning or classical conditioning) which achieves optimality through continuous identification of, and adaptation to, the causal relationship between respiratory motor output and resultant chemical mechanical afferent feedbacks. This internal model self-tuning adaptive control paradigm opens a new challenge and exciting opportunity for experimental and theoretical elucidations of the mechanisms of respiratory control - and of homeostatic regulation and sensorimotor integration in general. PMID- 17416558 TI - Reconsideration of sample pH adjustment in bioanalytical liquid-liquid extraction of ionisable compounds. AB - Liquid-liquid extraction (LLE) is widely used as a simple and robust sample preparation technique in bioanalytical sample preparation. When extracting ionisable compounds, most bioanalysts adjust the pH of the sample to achieve fully unionized compounds. Usually, a generally accepted rule is applied to adjust the pH of the aqueous phase, known as the pKa+/-2 rule, depending on the acid/base characteristics of the analyte. By taking a closer look at the general equations that describe the extraction behaviour of ionisable compounds, we extended this pH adjustment rule by taking the distribution ratio and the volume of both liquid phases into account. By choosing an extraction pH based on this extended rule, the selectivity of the extraction can be influenced without loss of recovery. As a measure of this selectivity, two equations were proposed to indicate the ability of the extraction system to discriminate between two compounds. Also, milder extraction pH can be used for pH labile analytes. To use this new rule quantitatively, a new calculation method for the determination of the distribution ratio was derived. These calculations were based on normalized recoveries making this method less susceptible to errors in absolute recovery determination. The proposed equations were supported by demonstrating that careful pH adjustment can lead to higher selectivity. The main conclusion was that a closer look at the extraction pH in bioanalytical methods extends the possibilities of obtaining a higher selectivity or the possibilities of extracting pH labile analytes at milder pH conditions without loss of recovery. PMID- 17416559 TI - One-step elimination of L-cysteine desulfhydrase from crude enzyme extracts of Pseudomonas sp. TS1138 using an immunomagnetic affinity matrix improves the enzymatic production of L-cysteine. AB - In this study, a high efficiency immunomagnetic affinity matrix was developed to eliminate L-cysteine desulfhydrase (CD), which decomposes L-cysteine, in crude enzyme extracts from Pseudomonas sp. TS1138. After cloning and expression in Escherichia coli, recombinant CD was purified to raise polyclonal antibodies from mice. The anti-CD antibody was cross-linked to staphylococcal protein A-magnetic cellulose microspheres (MCMS) with dimethyl pimelimidate (DMP). The natural CD was eliminated from the crude enzyme extracts by treatment with the cross-linked antibody-protein A-MCMS, resulting in a high level of L-cysteine production. The conversion rate of DL-2-amino-Delta2-thiazoline-4-carboxylic acid (DL-ATC) to L cysteine increased significantly from 61.9 to 96.2%. The cross-linked antibody protein A-MCMS showed its durability after repetitive use, maintaining a constant binding capacity for CD during five cycles. This study may lead to a convenient and cost-efficient method to produce L-cysteine by enzymatic conversions. PMID- 17416560 TI - Association between chronic hepatitis B virus infection and diabetes among Asian Americans and Pacific Islanders. AB - BACKGROUND: Asians have a higher prevalence of both diabetes (diabetes mellitus) and chronic hepatitis B virus infection compared to Caucasians. The aim of this study was to investigate whether hepatitis B virus infection was associated with diabetes mellitus among Asian Americans and Pacific Islanders. METHODS: We reviewed the electronic medical records of 411 Asian and 424 Pacific Islanders seen at our medical centre over a 5-year period. Diabetes mellitus was defined by the presence of two or more random blood glucose levels > or =200mg/dL, an ICD-9 diagnostic code of diabetes mellitus, or use of medications for diabetes mellitus. Hepatitis B virus infection was defined by a positive HBsAg test. RESULTS: Diabetes mellitus was diagnosed in 223 of the 835 subjects (26.7%), whereas hepatitis B virus infection was diagnosed in 56 (13.8%) of the 407 subjects tested for HBsAg. Overall, the prevalence of diabetes mellitus was significantly higher in patients with hepatitis B virus than in those without hepatitis B virus (58.9% vs. 33.3%, P<0.001), and this remained significant after adjustment for potential confounding variables (OR=3.17; 95% CI, 1.58-6.35). When Asians and Pacific Islanders were analysed separately, the prevalence of diabetes mellitus in patients with hepatitis B virus was significantly higher than in those without hepatitis B virus among Asians (65.0% vs. 27.5%, P<0.001) but not in Pacific Islanders (43.8% vs. 37.1%, P=0.60). Among the 390 subjects who were tested for both hepatitis B virus and hepatitis C virus, the prevalence of diabetes mellitus was 29.4% in uninfected subjects, 44.4% in patients with hepatitis B virus monoinfection, 47.2% in patients with hepatitis C virus monoinfection and 85.0% in patients with hepatitis B virus and hepatitis C virus coinfection (P<0.001). CONCLUSIONS: Hepatitis B virus infection is strongly associated with diabetes mellitus among Asian Americans, but not in Pacific Islanders, whereas hepatitis C virus infection was associated with diabetes mellitus in both ethnic groups. PMID- 17416561 TI - Derivative spectrophotometry in the visible region using absorbance versus log wavelength or wavenumber determination of cyanocobalamin in injection solutions. AB - Two approaches have been adopted to increase the steepness of the slopes and the sharpness of the curvature of an absorption curve in the visible region. These are recording (i) A (1cm) versus log wavelength or (ii) A (1cm) versus wavenumber. The computer program was tested by calculating the ratios of the first derivative optima for a Gaussian band, and proved that changing wavelength into log wavelength or wavenumber is effective to squeeze an absorption curve non linearly. Spectrophotometric identification of cyanocobalamin in the visible region was carried out using the ratios (Delta D1/D2) calculated for lambda, log lambda and nu scales. The ratios were highly reproducible and independent of concentration. First, second and fourth-order derivative determination of cyanocobalamin in injection solutions was performed in the visible region at 20040.08, 17452.01, (20040.08 - 17452.01) cm(-1) for D11(nu), D12(nu) Delta D1 (D11-D12) (nu), respectively, and at 18018.02 cm(-1) for D2(nu) and D4(nu). PMID- 17416562 TI - Optimization of levofloxacin analysis by RP-HPLC using multivariate calibration technique. AB - A rapid and sensitive reverse phase high performance liquid chromatographic (RP HPLC) method for the analysis of levofloxacin from bulk materials, dosage formulations and human serum is described. This isocratic method employs, a Nucleosil, C18 (10 microm, 25 cm x 0.46 cm) column with a mobile phase of water and acetonitrile (6:5), where in phosphoric acid was used to adjust the pH to 2.9 and propylparaben as an internal standard. Optimization of levofloxacin analysis was carried out using multivariate calibration technique and detector response was recorded at five different wave lengths. A linear response (r > 0.9999) was observed in the range of 40 to 10000 ng ml(-1). The method shows good recoveries, intra and inter-day relative standard deviations were less than 1.2 %. Validation parameters as specificity, accuracy and robustness were also determined. The method can conveniently be used for analysis of levofloxacin pharmacokinetic levels in human serum and pharmaceutical formulations. PMID- 17416563 TI - Enterococci vs coliforms as a possible fecal contamination indicator: baseline data for Karachi. AB - Fecal contamination of drinking water is the major cause of water borne illnesses. For long time coliforms are exploited as fecal contamination indicator. However, recent studies indicate low survival rate of coliforms in stress conditions, hence it's use as indicator of fecal pollution is being abandoned in many parts of the developed world. Implementation of such strategy demands availability of local data in the cities like Karachi. The present study provides a comparison between coliforms and enterococcal load and its variation in sewage samples collected (June, August and November, 2006) from eighteen towns of Karachi. All the diluted samples were selective media to obtain colony-forming units (CFU) mainly for coliforms and enterococci. The bacteria isolated were identified on the basis of conventional microbiological methods. Observations thus obtained were subjected to rigorous statistical analysis. The total load of enterococci was found in range of 1.27-8.47 X 10(7) as compared to coliforms (3.03-13.9 X 10(7)). However, segregation of data reveals greater inter town variability in CFU/ml both in coliforms and enterococci as suggested by their cumulative standard deviation +/-1.5 X 107. Furthermore, CFU/ml of both coliforms and enterococci also varies to variable scale when collected at different time intervals and at intra town level. Conclusively, the studies suggest high survival rate and lower variability of Enterococci compared to escherichia hence indicating its potential advantage to be used as fecal contamination indicator. PMID- 17416564 TI - A study of the digestive effect of dates on sucrose and starch. AB - A study of the digestive effect of dates on sucrose and starch has been carried out. Digestion of sucrose and dates and starch and dates at pH 7.4 results in the breakdown of both sucrose and starch to glucose and fructose. On the contrary, digestion of sucrose and starch alone at pH 7.4 does not cause any breakdown suggesting that dates do affect the breakdown of sucrose and starch in the intestine. The variations in the analytical results are of the order of 1-2% in terms of RSD. PMID- 17416565 TI - Serotonin-1A receptor responsiveness in stress and following adaptation to stress. AB - Stress is one of the environmental factors that may predispose psychiatric illness such as, depression. Stress may come from external environment in the form of stimuli such as heat, cold, loud noise and lack of oxygen. A deficiency of serotonin (5-hydroxytryptamine; 5-HT) is described in human depression. Parallel studies on experimental animals show that exposure to an uncontrollable stress inducing situation elicits behavioral deficits and increases serotonin metabolism in the brain. Stress-induced behavioral deficits and the increases of brain serotonin did not occur when the stress was administered repeatedly for 5 days, suggesting adaptation has occurred. The present study shows that responses to 8-hydroxy-2-(di-n-propylamino)tetraline (8-OH-DPAT), a selective 5-HT 1A agonist decreased following exposure to single stress and the decreases were normalized following adaptation to stress. The drug 8-OH-DPAT was also found to attenuate stress-induced behavioral deficits. The results are discussed in the context of stress-induced psychiatric disorder such as, depression and its treatment by 5-HT 1A agonist. PMID- 17416566 TI - Evaluation of Tectona grandis leaves for wound healing activity. AB - The frontal leaves of Tectona grandis (Verabinaceae) are widely used in the folklore for the treatment of various kinds of wounds, especially burn wound. The present study was carried out to evaluate the effect of hydrochloric extract of Tectona grandis on experimentally induced wounds in rats and compare the effects observed with a known wound healing agent, Aloe vera. The models selected were excision wound, incision wound, burn wound and dead space wound. A suitable gel formulation was selected for the application using cellophane membrane penetration. In the excision wound and burn wound models, animals treated with Tectona grandis leaf extract showed significant reduction in period of epithelisation and wound contraction 50%. In the incision wound model, a significant increase in the breaking strength was observed. Tectona grandis leaf extract treatment orally produced a significant increase in the breaking strength, dry weight and hydroxyproline content of the granulation tissue in dead space wound. It was concluded that Tectona grandis leaf extract applied topically (5% and 10% gel formulation) or administered orally (250 mg and 500 mg/kg body weight) possesses wound healing activity. PMID- 17416567 TI - Two triterpenes lupanone and lupeol isolated and identified from Tamarindus indica linn. AB - Tamarindus indica, a useful medicinal plant was subjected to phytochemical investigation. Two triterpenes (lupanone and lupeol) have been isolated from the leaves of this plant. Their structures were elucidated with the help of physico chemical methods and spectroscopic techniques. The lupanone and lupeol from this plant are being reported for the first time. PMID- 17416568 TI - Antitumor and antibacterial activity of ethylacetate extract of Ludwigia hyssopifolia linn and its active principle piperine. AB - An alkaloid constituent 1-[5-(1,3-benzodioxol-5-yl)-1-oxo-2,4 pentadienyl]piperidine, trivial name piperine was isolated from Ludwigia hyssopifolia Linn. (Family-Onagraceae). The ethylacetate extract of the plant and the isolated compound piperine were studied for antitumor and in vitro antibacterial activity. Ethylacetate extract showed 73.05 and 84.14% inhibition of Agrobacterium tumefaciens-induced crown gall tumor formation in potato disc. Piperine exhibited antitumor activity with IC50 value of 13.50 microg/disc. Both ethylacetate extract and piperine showed mild to moderate antibacterial activity against selected Gram-positive and Gram-negative bacteria. PMID- 17416569 TI - In vitro interactions of captopril with H2-receptor antagonists. AB - Captopril is effective in the treatment of hypertension of all grades of severity. H2-receptors antagonists block gastric acid secretion and some cardiovascular effects of histamine. In view of the fact that, simultaneous administration of both drugs may alter the antihypertensive effect of captopril, present paper deals with the in vitro availability studies of captopril in presence of commonly used H2-receptor antagonists like cimetidine, ranitidine and famotidine. In order to simulate various pH levels in GI tract and to find out the kinetics and energetics of captopril-H2-receptor antagonist interactions, these studies were carried out in buffers of pH 4, 7.4 and 9 at 37 degrees C and at elevated temperatures. These studies clearly indicate that most of the H2 receptor antagonists bind to captopril, forming charge-transfer complexes. As a result, the availability of captopril was affected by the concurrent administration of H2-receptor antagonists. Accordingly coadministration of both the drugs should be avoided. PMID- 17416570 TI - Activity of synergistic combination Amoxy-cassia against Salmonella. AB - The object of this study was to formulate new, cost effective anti-microbial combination for Multi Drug Resistant (MDR) Salmonella enterica Serover Typhi (SEST) based on the synergistic activity of amoxicillin with the aqueous fruit solution of Cassia fistula (CFF), a medicinal plant found in Pakistan which when tested alone have weak antimicrobial activity against blood isolates of MDR SEST. MIC of MDR SEST for amoxicillin and CFF alone was 750 microg/ml and 3750 microg/ml. The MIC of amoxicillin in combination with CFF was 23.4-187.5 microg/ml and 0.37-1.56 microg/ml for MDR and MDS SEST respectively. Fractional Inhibitory Concentration Index (FICI) using chequer-board titration suggested synergism for 80% MDR and MDS SEST tested, no antagonism observed. Time kill kinetics showed difference > log2 in CFU of CFF having sub-lethal amoxicillin concentrations. This novel combination is named as Amoxy-cassia. In vivo it is found to be non toxic at 1 gm/body weight of mice. PMID- 17416571 TI - Antibacterial and cytotoxic activity of the acetone extract of the flowers of Salvia sclarea and some natural products. AB - The aim of this study was to investigate the antibacterial and the cytotoxic activity of the acetone extract of the flowers of Salvia sclarea and of some natural products (sclareol, sclareolide and ambrox). The antibacterial and the cytotoxic activity were determined by the dilution method. Sclareolide, ambrox and sclareol demonstrated a good antibacterial activity against Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27950, Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 29212. The acetonic extract of the flowers of Salvia sclarea has a significant cytotoxic activity against Hep-2 cells. PMID- 17416572 TI - Spectrophotometric methods for the simultaneous analysis of meclezine hydrochloride and pyridoxine hydrochloride in bulk drug and pharmaceutical formulations. AB - Three new spectrophotometric procedures for the simultaneous determination of pyridoxine hydrochloride and meclezine hydrochloride are described. The first method depends on the application of simultaneous equation to resolve the interference due to spectral overlapping. The analytical signals were measured at 231 and 220 nm. Calibration graphs were established for 1 to 20 microGmL(-1) for pyridoxine hydrochloride and 0.5 to 10 microGmL(-1) for meclezine hydrochloride in binary mixture. In the second method, the determination of pyridoxine hydrochloride and meclezine hydrochloride was performed by measuring the absorbances at 290 and 235 nm in the simple absorbance spectra of their mixture. In third method a yellowish orange complex of pyridoxine hydrochloride was formed with ferric chloride, which absorbs in the visible region with lambda(max) at 445 nm. Calibration curve of complex formation range was conducted in between 20 to 250 microGmL(-1). These methods were validated with respect to accuracy, precision, linearity, limit of detection and quantification. Regression analysis of Beer's plot showed good correlation in a general concentration range of 1 to 20 microGml(-1) with correlation coefficient (r = 0.9999 and 0.9999; CV < 0.858) for pyridoxine hydrochloride, whereas meclezine hydrochloride concentration range 0.5 to 10 microGmL(-1) with correlation coefficient (r = 0.9998 and 0.9998; CV < 0.826). These methods can be readily applied, without any interference from the excipients. The suggested procedures were successfully applied to the determination of these compounds in synthetic mixtures and in pharmaceutical preparations, with high percentage of recovery, good accuracy and precision. PMID- 17416573 TI - Design and optimization of NSAID loaded nanoparticles. AB - The objective of the study was to design and evaluate NSAID loaded Nanoparticles drug delivery system, where Flurbiprofen (model drug) Nanoparticles with suitable size range are envisaged to concentrate at inflammation sites due to increase fragility of blood vessels at those sites and increased aggregation and prostaglandin synthesis. Materials used were surfactant (pluronic F 68) and polymer (poly lactic co glycolic acid; PLGA). The flurbiprofen loaded nanoparticles were prepared by solvent diffusion nano-precipitation method. Experiment was carried out following 3(2) factorial designs, where drug-polymer ratio was varied to optimize the formulation. From I.R studies no drug-polymer interaction was found. Particles size analysis was done using Malvern Mastersizer. Two parameters, namely, drug-polymer ratio and solvent-nonsolvent ratio were chosen for optimization following the factorial design. Amount of drug loading and surfactant were kept constant, and only polymer load was varied. The in-vitro drug release profile from nanoparticles was found to follow Higuchi square root kinetics implying a diffusion dependent release as is expected of an insoluble, non-swellable nature of PLGA. It indicated that nanoparticles formed were matrix in nature, in which flurbiprofen dispersed uniformly. Suitable polynomial models were generated and statistically validated using ANOVA for the different responses, namely drug release (maximization) and particle size (minimization). Those models were solved numerically and simultaneously to optimize the required formulation. Optimized formulation were found to have a polymer-drug ratio of 18.89:1 and manufactured at a nonsolvent-solvent ratio of 4:1 to maximized release after 8 hrs and minimized particle size. The methodology avoids the use of organic solvent and thus provides a safe, reproducible and fast method of production of nanoparticles. The study collaborates on the feasibility and suitability of aqueous polymeric drug delivery system, employing statistical design to develop a clinically useful Nanoparticle system with targeting potential. PMID- 17416574 TI - Multivariate calibrations in UV spectrophotometric analysis. AB - Calibration allows the user to relate instrumental measurements to the sample of interest. Multivariate calibration allows for the analysis of several measurements from several samples or specimens. The method contributes to the two steps procedure where step one involves the calibration of data and second step involves the prediction that are made or based on the calibration. In calibration, indirect measurements are made from samples where the amount of the analyte has been predetermined, usually by an independent assay or technique. These measurements, along with the predetermined analyte levels, comprise a group known as the calibration set. This set is used to develop a model that relates the amount of sample to the measurements by the instrument. In some cases, the construction of the model is simple due to a certain relationship, such as Beer's Law in the application of UV spectroscopy. Unlike spectroscopy, other cases can be much more complex, and it is in these cases where construction of the model is time-consuming step. Once the model is constructed, it can predict analyte levels based on measurements of new samples. It can be used to separate samples from interferences without the need of highly selective measurements for the analyte. Calibration techniques (used in the calibration step) differ in determining coefficient values for the preceding or similar equations. PMID- 17416575 TI - [Our experiences in treating patients with Hodgkin disease in the last decade]. AB - INTRODUCTION: Recently, in the diagnostics and treatment of Hodgkin's disease significant developments have occurred. AIM: To summarize the clinical and histological data of patients with Hodgkin's disease, treated at the 3rd Department of Internal Medicine, University of Debrecen between 1995-2004. In 2006 January, the mean follow-up was 69 (12-132) months. METHODS: Patients data was analyzed by using SPSS statistical software. RESULTS: The mean age of the 163 patients at the diagnosis was 36 years (14-75), with bimodal age distribution, the most frequent disease subtype was mixed-cell Hodgkin's disease (48.5%). 41.1% of the patients was at early stage, 15.7% had the worst prognosis, while 28.8% had bulky tumor. 7 patients had radiotherapy, 63 had chemotherapy, while at 92 patients combined modality treatment was used. 61.6% of radiotherapies were involved field, 61 patients received cyclophosphamide, vincristine, procarbazine, prednisolone, adriamycine, bleomycin, vinblastine, 87 adriamycine, bleomycin, vinblastine, 7 had other chemotherapies. As the response to the primary treatment 146 complete, 10 partial remission occurred, while 6 patients showed no response. 10 patients with partial remission and 5 non-responders were continually treated. 27 patients with complete remission had relapse, while 15 had high dose treatment with autologous peripheral stem cell transplantation. During the follow-up 18 patients died, 11 due to the lymphoma progression, or as the result of treatment, 6 had secondary malignancies, 1 due to other reasons. The 10-year prognosed overall survival was 83% (in details: early, advanced, favourable vs. unfavourable: 100% vs. 87.8%, 88.9% vs. 41.6%), the event free survival was 70% (82.6% vs. 70.8%, 64.5% vs. 0%). CONCLUSION: The treatment results of our Hodgkin's disease patients improved, additionally we showed that patients with early stage favourable disease the treatment toxicity should be reduced, while patients with advanced, unfavourable prognosis (10% of all patients) aggressive primary treatment should be used even with more severe side effects and complications. PMID- 17416576 TI - [Hyperglycemia and mortality in critically ill children]. AB - Several pathologic conditions are accompanied by stress-induced hyperglycemia in non-diabetic individuals which influences mortality and morbidity. AIMS: Prospective studies in adults support that glycemic control is an independent predictor of survival and normoglycemia has a beneficial effect on the outcome of patients. Few data are available in children, however a retrospective study documented significant correlation between mortality and duration, intensity and peak value of hyperglycemia. In the present study, the authors investigated the relationship between blood glucose level and pathological process in a multidisciplinary pediatric intensive care department retrospectively. RESULTS: It has been shown that highest blood glucose values were associated with fatal outcome independent of diagnosis (mean: 14,38 mmol/l) and with septicemia independent of final outcome (mean: 13,97 mmol/l). Patients with fatal outcome were hyperglycemic during the total duration (mean: 7,59 mmol/l) and on the last day (mean: 7,00 mmol/l) of treatment. Patients who survived had significantly lower blood glucose over the whole duration (mean: 6,52 mmol/l; p < 0,01) and on the last day of treatment (mean: 5,28 mmol/l; p < 0,01) than those who died. Percent of treatment days with blood glucose > = 6,1 mmol/l was significantly lower in case of survival as compared with fatal cases (42,68 vs. 74.07 %; p < 0,01) and the highest rate was observed in those with fatal septcemia (mean: 76,52 %). CONCLUSIONS: These data support that, similarly to adults, critical condition in children induces sustained hyperglycemia and higher peak values and longer duration of elevated blood glucose are associated with higher mortality rate. Septicemia proved to be potent inductor of abnormalities of carbohydrate metabolism. PMID- 17416577 TI - [Risk of recurrence in major craniospinal malformations between 1976 and 2005]. AB - INTRODUCTION: Craniospinal malformations are among the commonest developmental disorders. Due to its incidence the knowledge of the risk of recurrence is very important not only for the geneticist but also for the married couples. METHOD: In the period between January 1st, 1976 and December 31st, 2005, among the 75320 documented cases at the Genetic Counselling Units of the Departments of Obstetrics and Gynecology of the University of Debrecen and Semmelweis University Budapest, consultations were requested due to previous craniospinal malformations in the patients' history in 3030 cases (4.2%). This paper contains the most important informations about these cases of craniospinal malformations. The greatest risk of non-central nervous system developmental anomalies was associated with holoprosencephaly. RESULTS: In approximately two thirds of the cases (65.1%), the couples sought counselling because of malformation in a previous pregnancy. In these cases, the risk of recurrence was thought to be 5.2%, while in the case of two affected children this figure stood at 21.9%. When the risk of recurrence was analysed according to the various types, much lower figures (3.8%) were found for neural tube defects compared to other anomalies in this study. CONCLUSION: Analysing the values for the risk of recurrence in five year periods, neural tube defects (particularly anencephaly and spina bifida) showed detectable decrease, which could be attributed to a widening use of folic acid supplementation around about the time of conception and during pregnancy. PMID- 17416578 TI - [Astrocytes in health and disease]. AB - It is now known that astrocytes are not merely supporting cells but they also play an important role in neuronal functions. Astrocytes tightly ensheath neuronal synapses and regulate the excitation of neurons by the uptake of neurotransmitters; regulate the cerebral blood flow, cerebral fluid volume and extracellular concentrations of ions. They also supply fuel in the form of lactate and provide free radical scavengers such as glutathione for active neurons. These facts indicate that impaired function of astrocytes may lead to neuronal dysfunction. After brain injury (stroke, trauma or tumors) astrocytes are swollen and release active molecules such as glutamate or free radicals resulting in neuronal dysfunction. Thus, investigation of the molecular mechanisms of astrocyte function may reveal novel targets for the development of therapeutic tools in neuronal diseases. PMID- 17416579 TI - [Hungarian national dietary survey, 2003-2004. Micronutrients: mineral salts]. AB - In the course of the 3rd national nutritional survey data were collected in 2003, and the paper on the energy and nutrition intake of a sample group consisting of 1179 people all aged above 19, was published in the Hungarian Medical Journal (Vol. 146, No. 34, 2005). The present paper focuses on one group of micronutrients, namely the minerals' intake. The authors evaluated the results mainly in comparison with the data of the two previous Hungarian national surveys, and the international and national recommendations. The average magnesium intake was considered adequate in both sexes, while iron, zinc and copper intakes also met the recommended level in men. On the other hand, the insufficient calcium and the excessive phosphorus intakes, as well as the disproportionate calcium-phosphorus ratio in both sexes are unfavourable facts. The adverse results include insufficient iron, zinc and copper intake in women and the excessive sodium intake in both sexes, as well. The authors emphasize the importance of spreading the principles of healthy nutrition as well as co operating with the food industry in order to prevent the occurrence of mineral nutrients deficiency. PMID- 17416580 TI - [Celiac disease]. PMID- 17416582 TI - [6th Hungarian Futurology Conference, Gyor, October 6-7, 2006]. PMID- 17416585 TI - Allosteric regulation of SecA: magnesium-mediated control of conformation and activity. AB - In bacteria, the SecA protein associates with a ubiquitous protein channel SecYEG where it drives the post-translational secretion of pre-proteins across the plasma membrane. The high-resolution structures of both proteins have been determined in their resting states; however, the mechanism that couples ATP hydrolysis to active transport of substrate proteins through the membrane is not well understood. An analysis of the steady-state ATPase activity of the enzyme reveals that there is an allosteric binding site for magnesium distinct from that associated with hydrolysis of ATP. We have demonstrated that this regulation involves a large conformational change to the SecA dimer, which exerts a strong influence on the turnover and affinity for ATP, as well as the affinity for ADP. The strong inhibitory influence of magnesium on the ATPase activity can be countered by cardiolipin and conditions that promote protein translocation. PMID- 17416586 TI - FoxP3 enhances HIV-1 gene expression by modulating NFkappaB occupancy at the long terminal repeat in human T cells. AB - FoxP3 determines the development of CD4+CD25+ regulatory T (Treg) cells and represses interleukin-2 (IL-2) expression in Treg cells. However, human immunodeficiency virus type 1 (HIV-1) infects and replicates efficiently in FoxP3+ Treg cells. We report that, while inhibiting IL-2 gene expression, FoxP3 enhances gene expression from HIV-1 long terminal repeat (LTR). This FoxP3 activity requires both the N- and C-terminal domains and is inactivated by human IPEX (immunodysregulation, polyendocrinopathy, enteropathy, X-linked syndrome) mutations. FoxP3 enhances HIV-1 LTR via its specific NFkappaB binding sequences in an NFkappaB-dependent fashion in T cells but not in HEK293 cells. FoxP3 decreases level of histone acetylation at the interleukin-2 locus but not at the HIV-1 LTR. Although NFkappaB nuclear translocation is not altered, FoxP3 enhances NFkappaB-p65 binding to HIV-1 LTR. These data suggest that FoxP3 modulates gene expression in a promoter sequence-dependent fashion by modulating chromatin structure and NFkappaB activity. HIV-1 LTR has evolved to both highjack the T cell activation pathway for expression and to resist FoxP3-mediated suppression of T-cell activation. PMID- 17416587 TI - Site-specific mutations in HIV-1 gp41 reveal a correlation between HIV-1-mediated bystander apoptosis and fusion/hemifusion. AB - The loss of CD4(+) T cells in HIV-1 infections is hypothesized to be caused by apoptosis of bystander cells mediated by cell surface-expressed HIV-1 Env glycoprotein. However, the mechanism by which Env mediates this process remains controversial. Specifically, the role of HIV-1 gp120 binding to CD4 and CXCR4 versus the fusion process mediated by gp41 remains unresolved. Env-induced apoptosis in bystander cells has been shown to be gp41-dependent and correlates with the redistribution of membrane lipids between Env-expressing cells and target cells (hemifusion). Using a rational mutagenesis approach aimed at targeting Env function via the gp41 subunit, we examined the role of HIV gp41 in bystander apoptosis. A mutation in the fusion domain of gp41 (V513E) resulted in a fusion-defective Env that failed to induce apoptosis. A mutation in the gp41 N terminal helix (G547D) reduced cell fusion capacity and apoptosis; conversely, an Env mutant with a deletion of the gp41 cytoplasmic tail (Ct Del) enhanced both cell-to-cell fusion and apoptosis. Most significantly, an Env mutant containing a substitution in the loop region of gp41 (D589L) mediated transfer of lipids (hemifusion) to bystander cells but was defective in cell-to-cell and to a lesser degree virus-to-cell fusion. This mutant was still able to induce apoptosis in bystander cells. Hence, we have provided the first direct evidence that gp41 mediated hemifusion is both required and sufficient for induction of apoptosis in bystander cells. These results may help to explain the mechanism of HIV-1 Env induced T cell depletion. PMID- 17416588 TI - ADAM-15/metargidin mediates homotypic aggregation of human T lymphocytes and heterotypic interactions of T lymphocytes with intestinal epithelial cells. AB - Intestinal epithelial cells (IEC) play an immunoregulatory role in the intestine. This role involves cell-cell interactions with intraepithelial lymphocytes that may also play a role in some enteropathies. The discovery of the RGD motif containing Protein ADAM-15 (a disintegrin and metalloprotease-15) raises the question of its involvement in these cell-cell interactions. Cell adhesion assays were performed using the Jurkat E6.1 T cell line as a model of T lymphocytes and Caco2-BBE monolayers as a model of intestinal epithelia. Our results show that an anti-ADAM-15 ectodomain antibody inhibited the attachment of Jurkat cells on Caco2-BBE monolayers. Overexpression of ADAM-15 in Caco2-BBE cells enhanced Jurkat cell binding, and overexpression of ADAM-15 in Jurkat cells enhanced their aggregation. Mutagenesis experiments showed that both the mutation of ADAM-15 RGD domain or the deletion of its cytoplasmic tail decreased these cell-cell interactions. Moreover, wound-healing experiments showed that epithelial ADAM-15 mediated Jurkat cell adhesion to Caco2-BBE cells enhances the mechanisms of wound repair. We also found that ADAM-15-mediated aggregation of Jurkat cells increases the expression of tumor necrosis factor-alpha mRNA. These results demonstrate the following: 1) ADAM-15 is involved in heterotypic adhesion of intraepithelial lymphocytes to IEC as well as in homotypic aggregation of T cells; 2) both the RGD motif and the cytoplasmic tail of ADAM-15 are involved for these cell-cell interactions; and 3) ADAM-15-mediated cell-cell interactions are involved in mechanisms of epithelial restitution and production of pro-inflammatory mediators. Altogether these findings point to ADAM-15 as a possible therapeutic target for prevention of inappropriate T cell activation involved in some pathologies. PMID- 17416589 TI - Genetic evidence supporting caveolae microdomain regulation of calcium entry in endothelial cells. AB - Various cellular signals initiate calcium entry into cells, and there is evidence that lipid rafts and caveolae may concentrate proteins that regulate transmembrane calcium fluxes. Here, using mice deficient in caveolin-1 (Cav-1) and Cav-1 knock-out reconstituted with endothelium-specific Cav-1, we show that Cav-1 is essential for calcium entry in endothelial cells and governs the localization and protein-protein interactions between transient receptor channels C4 and C1. Thus, Cav-1 is required for calcium entry in vascular endothelial cells and perhaps other specialized cell types containing caveolae. PMID- 17416590 TI - Thrombospondin-1 inhibits nitric oxide signaling via CD36 by inhibiting myristic acid uptake. AB - Although CD36 is generally recognized to be an inhibitory signaling receptor for thrombospondin-1 (TSP1), the molecular mechanism for transduction of this signal remains unclear. Based on evidence that myristic acid and TSP1 each modulate endothelial cell nitric oxide signaling in a CD36-dependent manner, we examined the ability of TSP1 to modulate the fatty acid translocase activity of CD36. TSP1 and a CD36 antibody that mimics the activity of TSP1 inhibited myristate uptake. Recombinant TSP1 type 1 repeats were weakly inhibitory, but an anti-angiogenic peptide derived from this domain potently inhibited myristate uptake. This peptide also inhibited membrane translocation of the myristoylated CD36 signaling target Fyn and activation of Src family kinases. Myristate uptake stimulated cGMP synthesis via endothelial nitric-oxide synthase and soluble guanylyl cyclase. CD36 ligands blocked myristate-stimulated cGMP accumulation in proportion to their ability to inhibit myristate uptake. TSP1 also inhibited myristate stimulated cGMP synthesis by engaging its receptor CD47. Myristate stimulated endothelial and vascular smooth muscle cell adhesion on type I collagen via the NO/cGMP pathway, and CD36 ligands that inhibit myristate uptake blocked this response. Therefore, the fatty acid translocase activity of CD36 elicits proangiogenic signaling in vascular cells, and TSP1 inhibits this response by simultaneously inhibiting fatty acid uptake via CD36 and downstream cGMP signaling via CD47. PMID- 17416591 TI - Different modes of binding of mono- and biaromatic effectors to the transcriptional regulator TTGV: role in differential derepression from its cognate operator. AB - Members of the IclR family of regulators exhibit a highly conserved effector recognition domain and interact with a limited number of effectors. In contrast with most IclR family members, TtgV, the transcriptional repressor of the TtgGHI efflux pump, exhibits multidrug recognition properties. A three-dimensional model of the effector domain of TtgV was generated based on the available three dimensional structure of several IclR members, and a series of point mutants was created. Using isothermal titration calorimetry, we determined the binding parameters of the most efficient effectors for TtgV and its mutant variants. All mutants bound biaromatic compounds with higher affinity than the wild-type protein, whereas monoaromatic compounds were bound with lower affinity. This tendency was particularly pronounced for mutants F134A and H200A. TtgVF134A bound 4-nitrotoluene with an affinity 13-fold lower than that of TtgV (17.4+/-0.6 microM). This mutant bound 1-naphthol with an affinity of 5.7 microM, which is seven times as great as that of TtgV (40 microM). The TtgVV223A mutant bound to DNA with the same affinity as the wild-type TtgV protein, but it remained bound to the target operator in the presence of effectors, suggesting that Val-223 could be part of an intra-TtgV signal recognition pathway. Thermodynamic analyses of the binding of effectors to TtgV and to its mutants in complex with their target DNA revealed that the binding of biaromatic compounds resulted in a more efficient release of the repressor protein than the binding of monoaromatics. The physiological significance of these findings is discussed. PMID- 17416592 TI - Further studies in the reduction of residual in Harshaw TLD-100H (LiF:Mg,Cu,P). AB - This paper describes further investigations and results in the reduction of residual signal in Harshaw TLD-100H, 600H and 700H (LiF:Mg,Cu,P). TLD-100H is an advanced, relatively new dosimetric material with near tissue-equivalence, flat energy response, and the ability to measure beta, photon and, more importantly, neutrons all from the same base material. The simple glow curve structure provides insignificant fade over extended dosimetric periods of up to 1 y. A criticism of the material has been the residual as compared with TLD-100 (LiF:Mg,Ti). We will show how high-temperature peaks cause the residual signal. We will also show how the various parameters and conditions of the residual measurement technique as well as the configuration of the sample affect the residual measurements. A brief description of the experimental paths taken during our investigation will be presented. We will show how we have reduced the high temperature peaks of LiF:Mg,Cu,P in our manufacturing process while not affecting other dosimetric properties of this material. The improvements and material properties that need to be shown have been incorporated into our production processes. LiF:Mg,Cu,P stands as a premier choice for personal dosimetry and has been integrated into personal, environmental and extremity configurations of the Harshaw TLD family-line of products. PMID- 17416593 TI - Characterization of neutron field in a NPP workplace. AB - At the Krsko Nuclear Power Plant (NPP), albedo dosimeters are used for personal neutron dosimetry. Spectrometric measurements allow determination of reference dosimetric values of realistic neutron fields to be used for calibration of albedo dosimeters. The Laboratory for Neutron Metrology and Dosimetry from the Institute for Radiological Protection and Nuclear Safety (IRSN) was in charge of characterising neutron fields in the plant at two representative points with high neutron and gamma dose rate. Calibration of the dosimeters in the workplace used to be performed only by a spherical survey meter. Based on the reference dosimetric values, the Plant Dosimetry Laboratory has verified the response of albedo dosimeters. PMID- 17416594 TI - Chimeric structural stabilities in the coiled-coil structure of the NECK domain in human lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1). AB - LOX-1 (lectin-like oxidized low-density lipoprotein receptor 1) is the major oxidized LDL (OxLDL) receptor on endothelial cells. The extracellular part of LOX 1 comprises an 82-residue stalk region (NECK) and a C-type lectin-like ligand binding domain (CTLD). The NECK displays sequence similarity to the coiled-coil region of myosin, having been suggested it adopts a rod-like structure. In this article, we report the structural analyses of human LOX-1 NECK using a variety of approaches including limited proteolysis, chemical cross-linking, circular dichroism (CD) and NMR. Our analysis reveals a unique structural feature of the LOX-1 NECK. Despite significant sequence similarity with the myosin coiled-coil, LOX-1 NECK does not form a uniform rod-like structure. Although not random, one third of the N-terminal NECK is less structured than the remainder of the protein and is highly sensitive to cleavage by a variety of proteases. The coiled-coil structure is localized at the C-terminal part of the NECK, but is in dynamic equilibrium among multiple conformational states on a mus-ms time scale. This chimeric structural property of the NECK region may enable clustered LOX-1 on the cell surface to recognize OxLDL. PMID- 17416595 TI - Crucial roles for chromatin dynamics in cellular memory. AB - Cellular memory is defined as a long-term maintenance of a particular pattern of gene expression through many rounds of cell division or even after cell division. It is critical for development and differentiation of multicellular organisms. Chromatin dynamics including histone modification, histone replacement and chromatin remodeling play key roles in cellular memory. PMID- 17416596 TI - Angiotensin II-induced sudden arrhythmic death and electrical remodeling. AB - Rats harboring the human renin and angiotensinogen genes (dTGR) feature angiotensin (ANG) II/hypertension-induced cardiac damage and die suddenly between wk 7 and 8. We observed by electrocardiogram (ECG) telemetry that ventricular tachycardia (VT) is a common terminal event in these animals. Our aim was to investigate electrical remodeling. We used ECG telemetry, noninvasive cardiac magnetic field mapping (CMFM) at wk 5 and 7, and performed in vivo programmed electrical stimulation at wk 7. We also investigated whether or not losartan (Los; 30 mg x kg(-1) x day(-1)) would prevent electrical remodeling. Cardiac hypertrophy and systolic blood pressure progressively increased in dTGR compared with Sprague-Dawley (SD) controls. Already by wk 5, untreated dTGR showed increased perivascular and interstitial fibrosis, connective tissue growth factor expression, and monocyte infiltration compared with SD rats, differences that progressed through time. Left-ventricular mRNA expression of potassium channel subunit Kv4.3 and gap-junction protein connexin 43 were significantly reduced in dTGR compared with Los-treated dTGR and SD. CMFM showed that depolarization and repolarization were prolonged and inhomogeneous. Los ameliorated all disturbances. VT could be induced in 88% of dTGR but only in 33% of Los-treated dTGR and could not be induced in SD. Untreated dTGR show electrical remodeling and probably die from VT. Los treatment reduces myocardial remodeling and predisposition to arrhythmias. ANG II target organ damage induces VT. PMID- 17416597 TI - Pulmonary arterial hypertension: a disease of tethers, SNAREs and SNAPs? AB - Histological and electron microscopic studies over the past four decades have highlighted "plump," "enlarged" endothelial, smooth muscle, and fibroblastic cellular elements with increased endoplasmic reticulum, Golgi stacks, and vacuolation in pulmonary arterial lesions in human and in experimental (hypoxia and monocrotaline) pulmonary arterial hypertension. However, the contribution of disrupted intracellular membrane trafficking in the pathobiology of this disease has received insufficient attention. Recent studies suggest a pathogenetic role of the disruption of intracellular trafficking of vasorelevant proteins and cell surface receptors in the development of this disease. The purpose of this essay is to highlight the molecular regulation of vesicular trafficking by membrane tethers, SNAREs and SNAPs, and to suggest how their dysfunction, directly and/or indirectly, might contribute to development of pulmonary arterial hypertension in experimental models and in humans, including that due to mutations in bone morphogenetic receptor type 2. PMID- 17416598 TI - Human recombinant chromogranin A-derived vasostatin-1 mimics preconditioning via an adenosine/nitric oxide signaling mechanism. AB - The acidic protein chromogranin A (CgA) is the precursor of several regulatory peptides generated by specific proteolytic processes. Human recombinant CgA NH(2) terminal fragment STA-CgA(1-78) (hrSTA-CgA(1-78)), containing vasostatin-1 (CgA(1 76)) domain, exerts a negative inotropic effect and counteracts the beta adrenergic positive inotropic effect on the rat heart. We hypothesized an involvement of nitric oxide (NO)-dependent pathway in both cardiodepression and cardioprotection by hrSTA-CgA(1-78). We also hypothesized an involvement of adenosine A(1) receptor and protein kinase C (PKC) in cardioprotection by hrSTA CgA(1-78). Therefore, we evaluated whether 1) the cardioinhibition mediated by hrSTA-CgA(1-78) involves the G(i/o) proteins/NO-dependent signal transduction cascade, 2) hrSTA-CgA(1-78) induces ischemic preconditioning-like protective effects on the myocardium, and 3) inhibition of NO synthase (NOS), adenosine A(1) receptor, or PKC affects hrSTA-CgA(1-78) protection. Using the isolated rat heart, we found that the reduction of left ventricular pressure (LVP), rate pressure product, and maximal values of the first derivative of LVP elicited by hrSTA-CgA(1-78) at 33 nM is abolished by blocking G(i/o) proteins with pertussis toxin, scavenging NO with hemoglobin, and blocking NOS activity with N(G) monomethyl-l-arginine or N(5)-(iminoethyl)-l-ornithine, soluble guanylate cyclase with 1H-[1,2,4]oxadiazole-[4,4-a]quinoxalin-1-one, and protein kinase (PKG) with KT5823. Data suggest the involvement of the G(i/o) proteins/NO-cGMP-PKG pathway in the hrSTA-CgA(1-78)-dependent cardioinhibition. When given before 30 min of ischemia, hrSTA-CgA(1-78) significantly reduced the size of the infarct from 64 +/- 4 to 32 +/- 3% of the left ventricular mass. This protective effect was abolished by either NOS inhibition or PKC blockade and was attenuated, but not suppressed, by the blockade of A(1) receptors. These results suggest that hrSTA CgA(1-78) activity triggers two different pathways: one of these pathways is mediated by A(1) receptors, and the other is mediated by NO release. As with repeated brief preconditioning ischemia, hrSTA-CgA(1-78) may be considered a stimulus strong enough to trigger both pathways, which may converge on PKC. PMID- 17416599 TI - Increased mitochondrial H2O2 production promotes endothelial NF-kappaB activation in aged rat arteries. AB - Previous studies have shown that the aging vascular system undergoes pro atherogenic phenotypic changes, including increased oxidative stress and a pro inflammatory shift in endothelial gene expression profile. To elucidate the link between increased oxidative stress and vascular inflammation in aging, we compared the carotid arteries and aortas of young and aged (24 mo old) Fisher 344 rats. In aged vessels there was an increased NF-kappaB activity (assessed by luciferase reporter gene assay and NF-kappaB binding assay), which was attenuated by scavenging H(2)O(2). Aging did not alter the vascular mRNA and protein expression of p65 and p50 subunits of NF-kappaB. In endothelial cells of aged vessels there was an increased production of H(2)O(2) (assessed by 5,6 chloromethyl-2',7'-dichlorodihydrofluorescein diacetate-acetyl ester fluorescence), which was attenuated by the mitochondrial uncoupler FCCP. In young arteries and cultured endothelial cells, antimycin A plus succinate significantly increased FCCP-sensitive mitochondrial H(2)O(2) generation, which was associated with activation of NF-kappaB. In aged vessels inhibition of NF-kappaB (by pyrrolidenedithiocarbamate, resveratrol) significantly attenuated inflammatory gene expression and inhibited monocyte adhesiveness. Thus increased mitochondrial oxidative stress contributes to endothelial NF-kappaB activation, which contributes to the pro-inflammatory phenotypic alterations in the aged vaculature. Our model predicts that by reducing mitochondrial H(2)O(2) production and/or directly inhibiting NF-kappaB novel anti-aging pharmacological treatments (e.g., calorie restriction mimetics) will exert significant anti-inflammatory and vasoprotective effects. PMID- 17416600 TI - Rescue of tropomyosin-induced familial hypertrophic cardiomyopathy mice by transgenesis. AB - Familial hypertrophic cardiomyopathy (FHC) is a disease caused by mutations in contractile proteins of the sarcomere. Our laboratory developed a mouse model of FHC with a mutation in the thin filament protein alpha-tropomyosin (TM) at amino acid 180 (Glu180Gly). The hearts of these mice exhibit dramatic systolic and diastolic dysfunction, and their myofilaments demonstrate increased calcium sensitivity. The mice also develop severe cardiac hypertrophy, with death ensuing by 6 mo. In an attempt to normalize calcium sensitivity in the cardiomyofilaments of the hypertrophic mice, we generated a chimeric alpha-/beta-TM protein that decreases calcium sensitivity in transgenic mouse cardiac myofilaments. By mating mice from these two models together, we tested the hypothesis that an attenuation of myofilament calcium sensitivity would modulate the severe physiological and pathological consequences of the FHC mutation. These double-transgenic mice "rescue" the hypertrophic phenotype by exhibiting a normal morphology with no pathological abnormalities. Physiological analyses of these rescued mice show improved cardiac function and normal myofilament calcium sensitivity. These results demonstrate that alterations in calcium response by modification of contractile proteins can prevent the pathological and physiological effects of this disease. PMID- 17416601 TI - Transplanted embryonic stem cells following mouse myocardial infarction inhibit apoptosis and cardiac remodeling. AB - We have previously shown that mouse embryonic stem (ES) cells transplanted following myocardial infarction (MI) differentiate into the major cell types in the heart and improve cardiac function. However, the extent of regeneration was relatively meager compared with the observed functional improvement. Therefore, we hypothesize that mechanisms in addition to regeneration contribute to the functional improvement from ES cell therapy. In this study, we examined the effect of mouse ES cells transplanted post-MI on cardiac apoptosis, fibrosis, and hypertrophy. MI was produced by left coronary artery ligation in C57BL/6 mice. Two different mouse ES cell lines, expressing enhanced green fluorescent protein and beta-galactosidase, respectively, were tested. Post-MI intramyocardial injection of 3 x 10(4) ES cells was compared with injection of medium alone. Terminal deoxynucleotidyl nick end labeling (TUNEL), immunofluorescence, and histology were used to examine the effect of transplanted ES cells on apoptosis, fibrosis, and hypertrophy. Two weeks post-MI, ES cell-transplanted hearts exhibited a significant decrease in TUNEL-stained nuclei (mean +/- SE; MI+medium = 12 +/- 1.5%; MI+ES cells = 6.6 +/- 1%, P < 0.05). TUNEL-positive nuclei were confirmed to be apoptotic by colabeling with a caspase-3 antibody. Cardiac fibrosis was 57% less in the MI+ES cell group compared with the MI + medium group (P < 0.05) as shown with Masson's trichrome staining. Picrosirius red staining confirmed a decreased amount of collagen present in the MI+ES cell group. Cardiomyocyte hypertrophy was significantly decreased following ES cell transplantation compared with medium control animals. In conclusion, transplanted mouse ES cells in the infarcted heart inhibit apoptosis, fibrosis, and hypertrophy, thereby reducing adverse remodeling. PMID- 17416602 TI - Cardiomyocyte-restricted restoration of nitric oxide synthase 3 attenuates left ventricular remodeling after chronic pressure overload. AB - Although nitric oxide synthase (NOS)3 is implicated as an important modulator of left ventricular (LV) remodeling, its role in the cardiac response to chronic pressure overload is controversial. We examined whether selective restoration of NOS3 to the hearts of NOS3-deficient mice would modulate the LV remodeling response to transverse aortic constriction (TAC). LV structure and function were compared at baseline and after TAC in NOS3-deficient (NOS3(-/-)) mice and NOS3(-/ ) mice carrying a transgene directing NOS3 expression specifically in cardiomyocytes (NOS3(-/-TG) mice). At baseline, echocardiographic assessment of LV dimensions and function, invasive hemodynamic measurements, LV mass, and myocyte width did not differ between the two genotypes. Four weeks after TAC, echocardiographic and hemodynamic indexes of LV systolic function indicated that contractile performance was better preserved in NOS3(-/-TG) mice than in NOS3(-/ ) mice. Echocardiographic LV wall thickness and cardiomyocyte width were greater in NOS3(-/-) mice than in NOS3(-/-TG) mice. TAC-induced cardiac fibrosis did not differ between these genotypes. TAC increased cardiac superoxide generation in NOS3(-/-TG) but not NOS3(-/-) mice. The ratio of NOS3 dimers to monomers did not differ before and after TAC in NOS3(-/-TG) mice. Restoration of NOS3 to the heart of NOS3-deficient mice attenuates LV hypertrophy and dysfunction after TAC, suggesting that NOS3 protects against the adverse LV remodeling induced by prolonged pressure overload. PMID- 17416603 TI - Rho kinase is an effector underlying Ca2+-desensitizing hypoxic relaxation in porcine coronary artery. AB - Acute hypoxia dilates most systemic arteries leading to increased tissue perfusion. We have previously shown that at high-stimulus conditions, porcine coronary artery was relaxed by hypoxia without a change in intracellular [Ca(2+)] (27). This Ca(2+)-desensitizing hypoxic relaxation (CDHR) was validated in permeabilized porcine coronary artery smooth muscle (PCASM) in which hypoxia decreased force and myosin regulatory light chain phosphorylation (p-MRLC) despite fixed [Ca(2+)] (10). Rho kinase-dependent phosphorylation of myosin phosphatase-targeting subunit 1 (p-MYPT1) is associated with decreased MRLC phosphatase activity and increased Ca(2+) sensitivity of both p-MRLC and force. We recently reported that p-MYPT1 dephosphorylation was a key effector in CDHR (33). In the current study, we tested the hypothesis that Rho kinase and not p MYPT1 phosphatase is the regulated enzyme involved in CDHR. We used alpha-toxin to permeabilize deendothelialized PCASM. CDHR was attenuated in contractions attributable to myosin light chain kinase (MLCK, in the presence of the Rho kinase inhibitor Y-27632). In contrast, hypoxia relaxed contractions attributable to Rho kinase phosphorylation of MYPT1 and MRLC or MRLC alone (in the presence of the MLCK inhibitor ML7). Using an in situ assay, we showed that Rho kinase activity, measured as thiophosphorylation of MYPT1 and MRLC, was nearly abolished by hypoxia. The in vitro activity of the catalytically active fragment of Rho kinase was not affected by hypoxia. Our evidence strongly implicates that hypoxia directly inhibits Rho kinase-dependent phosphorylation of MYPT1. This underlies the decreases in both p-MYPT1 and p-MRLC and thereby leads to the Ca(2+) desensitizing hypoxic relaxation. PMID- 17416604 TI - Enhanced activity of ventricular Na+-HCO3- cotransport in pressure overload hypertrophy. AB - The Na(+)-HCO(3)(-) cotransporter (NBC) plays a key role in intracellular pH (pH(i)) regulation in normal ventricular muscle. However, the state of NBC in nonischemic hypertrophied hearts is unresolved. In this study, we examined functional and molecular properties of NBC in adult rat ventricular myocytes. The cells were enzymatically isolated from both normal and hypertrophied hearts. Ventricular hypertrophy was induced by pressure overload created by suprarenal abdominal aortic constriction of 50% for 7 wk. pH(i) was measured in single cells using the fluorescent pH indicator 2',7'-bis(2-carboxyethyl)5 (6)carboxyfluorescein. Real-time PCR analysis was used to quantitatively assess expression of NBC-encoding mRNA, including SLC4A4 (encoding electrogenic NBC, NBCe1) and SLC4A7 (electroneutral NBC, NBCn1). Our results demonstrate that: 1) mRNA levels of both the electrogenic NBCe1 (SLC4A4) and electroneutral NBCn1 (SLC4A7) forms of NBC were increased by aortic constriction, 2) the onset of NBC upregulation occurred within 3 days after constriction, 3) normal and hypertrophied ventricles displayed regional differences in NBC expression, 4) acid extrusion via NBC (J(NBC)) was increased significantly in hypertrophied myocytes, 5) although acid extrusion via Na(+)/H(+) exchange was also increased in hypertrophied myocytes, the relative enhancement of J(NBC) was larger, 6) membrane depolarization markedly increased J(NBC) in hypertrophied myocytes, and 7) losartan, an ANG II AT(1) receptor antagonist, significantly attenuated the upregulation of both NBCs induced by 3 wk of aortic constriction. Enhanced NBC activity during hypertrophic development provides a mechanism for intracellular Na(+) overload, which may render the ventricles more vulnerable to Ca(2+) overload during ischemia-reperfusion. PMID- 17416605 TI - TNF-alpha blockade decreases oxidative stress in the paraventricular nucleus and attenuates sympathoexcitation in heart failure rats. AB - Oxidative stress plays an important role in the pathophysiology of cardiovascular disease. Recent evidence suggests that cytokines induce oxidative stress and contribute to cardiac dysfunction. In this study, we investigated whether increased circulating and tissue levels of tumor necrosis factor (TNF)-alpha in congestive heart failure (CHF) modulate the expression of NAD(P)H oxidase subunits, Nox2 and its isoforms, in the paraventricular nucleus (PVN) of the hypothalamus and contribute to exaggerated sympathetic drive in CHF. Heart failure was induced in Sprague-Dawly rats by coronary artery ligation and was confirmed using echocardiography. Pentoxifylline (PTX) was used to block the production of cytokines for a period of 5 wk. CHF induced a significant increase in the production of reactive oxygen species (ROS) in the left ventricle (LV) and in the PVN. The mRNA and protein expression of TNF-alpha, Nox1, Nox2, and Nox4 was significantly increased in the LV and PVN of CHF rats. CHF also decreased ejection fraction, increased Tei index, and increased circulating catecholamines (epinephrine and norepinephrine) and renal sympathetic activity (RSNA). In contrast, treatment with PTX in CHF rats completely blocked oxidative stress and decreased the production of TNF-alpha and Nox2 isoforms both in the LV and PVN. PTX treatment also decreased catecholamines and RSNA and prevented further decrease in cardiac function. In summary, TNF-alpha blockade attenuates ROS and sympathoexcitation in CHF. This study unveils new mechanisms by which cytokines play a role in the pathogenesis of CHF, thus underscoring the importance of targeting cytokines in heart failure. PMID- 17416606 TI - Repairing the myofilaments to heal the heart. PMID- 17416607 TI - Enhanced cardioprotection against ischemia-reperfusion injury with a dipyridamole and low-dose atorvastatin combination. AB - Atorvastatin (ATV) limits infarct size (IS) by activating Akt and ecto-5 nucleotidase, which generates adenosine. Activated Akt and adenosine activate endothelial nitric oxide synthase (eNOS). When given orally, high doses (10 mg/kg) are needed to achieve full protection. We determined whether dipyridamole (DIP), by preventing the reuptake of adenosine, has a synergistic effect with ATV in reducing myocardial IS. In this study, rats received 3-days of the following: water, ATV (2 mg.kg(-1).day(-1)), DIP (6 mg.kg(-1).day(-1)), or ATV + DIP. In addition, rats received 3-days of the following: aminophylline (Ami; 10 mg.kg( 1).day(-1)) or Ami + ATV + DIP. Rats underwent 30 min of myocardial ischemia followed by 4 h of reperfusion (IS protocol), or hearts were explanted for immunoblotting. As a result, IS in the controls was 34.0 +/- 2.8% of the area at risk. ATV (33.1 +/- 2.1%) and DIP (30.5 +/- 1.5%) did not affect IS, whereas ATV + DIP reduced IS (12.2 +/- 0.5%; P < 0.001 vs. each of the other groups). There was no difference in IS between the Ami alone (48.1 +/- 0.8%) and the Ami + ATV + DIP (45.8 +/- 2.9%) group (P = 0.422), suggesting that Ami completely blocked the protective effect. Myocardial adenosine level in the controls was 30.6 +/- 3.6 pg/microl. ATV (51.0 +/- 4.9 pg/microl) and DIP (51.5 +/- 6.8 pg/microl) caused a small increase in adenosine levels, whereas ATV + DIP caused a greater increase in adenosine levels (66.4 +/- 3.1 pg/microl). ATV and DIP alone did not affect myocardial Ser473 phosphorylated-Akt and Ser1177 phosphorylated-eNOS levels, whereas ATV + DIP significantly increased them. In conclusion, low-dose ATV and DIP had synergistic effects in reducing myocardial IS and activation of Akt and eNOS. This combination may have a potential benefit in augmenting the eNOS mediated pleiotropic effects of statins. PMID- 17416608 TI - Tumor necrosis factor-alpha is toxic via receptor 1 and protective via receptor 2 in a murine model of myocardial infarction. AB - Tumor necrosis factor (TNF)-alpha induced in damaged myocardium has been considered to be cardiotoxic. TNF-alpha initiates its biological effects by binding two distinct receptors: R1 (p55) and R2 (p75). Although TNF-alpha has been shown to be cardiotoxic via R1-mediated pathways, little is known about the roles of R2-mediated pathways in myocardial infarction (MI). We created MI in R1 knockout (R1KO), R2KO, and wild-type (WT) mice by ligating the left coronary artery. Functional, histological, and biochemical analyses were performed 4 wk after ligation. Although infarct size was not different among WT, R1KO, and R2KO mice, post-MI survival was significantly improved in R1KO but not R2KO mice. R1KO significantly ameliorated contractile dysfunction after MI, whereas R2KO significantly exaggerated ventricular dilatation and dysfunction. Myocyte hypertrophy and interstitial fibrosis in noninfarct myocardium was exacerbated in R2KO but not in R1KO mice. Expression of R1, which was not affected by MI and was nullified in R1KO mice, was significantly upregulated in R2KO mice. In contrast, expression of R2, which was significantly upregulated by MI and was nullified in R2KO mice, was unaffected in R1KO mice. Meanwhile, TNF-alpha expression, which was significantly upregulated in noninfarct myocardium after MI, was not affected by R1KO or R2KO. However, transcript levels of IL-6, IL-1beta, transforming growth factor-beta, and monocyte chemotactic protein-1, which were significantly upregulated after MI, were significantly downregulated in R1KO mice. In contrast, transcript levels of IL-6 and IL-1beta were significantly further upregulated in R2KO mice. TNF-alpha is toxic via R1 and protective via R2 in a murine model of MI. Selective blockade of R1 may be a candidate therapeutic intervention for MI. PMID- 17416609 TI - Rho kinase-mediated local cold-induced cutaneous vasoconstriction is augmented in aged human skin. AB - Cutaneous vasoconstriction (VC), a critical thermoregulatory response to cold, is generally impaired with aging. However, the effects of aging on local cooling induced VC and its underlying mechanisms are poorly understood. We tested whether aged skin exhibits attenuated localized cold-induced VC and whether Rho kinase mediated cold-induced VC is augmented with age. Skin blood flow was monitored with laser Doppler flowmetry (LDF) on seven young and seven older subjects. Cutaneous vascular conductance (CVC; LDF/mean arterial pressure) was expressed as percentage change from baseline (%DeltaCVC(base)). In protocol 1, two forearm skin sites were cooled to six temperatures (31.5-19 degrees C) for 10 min each or two temperatures (29 degrees C, 24 degrees C) for 30 min each, with no age differences in the magnitude of VC. In protocol 2, three forearm skin sites were instrumented for intradermal microdialysis and cooled to 24 degrees C for 40 min. During minutes 1-5, there was no age difference in CVC responses at control sites (young: -45 +/- 6% vs. older: -46 +/- 3%, P > 0.9). Adrenoceptor antagonism (yohimbine + propranolol) abolished VC in young (to +15 +/- 13%, P < 0.05) but only partially inhibited VC in older subjects (to -23 +/- 6%, P < 0.05). Rho kinase inhibition plus adrenoceptor antagonism (yohimbine + propranolol + fasudil) abolished VC in both groups. During minutes 35-40, there was no age difference in control (young: -77 +/- 4% vs. older: -70 +/- 2%, P > 0.3) or adrenoceptor-antagonized responses (young: -61 +/- 3% vs. older: -55 +/- 2%, P > 0.3); however, Rho kinase inhibition plus adrenoceptor antagonism blocked more VC in older compared with young subjects (-19 +/- 11% vs. -35 +/- 3%, P < 0.05). Although its magnitude remains unaffected, cold-induced VC becomes less dependent on adrenergic and more dependent on Rho kinase signaling with advancing age. PMID- 17416610 TI - Phosphorylation of human inhibitor-1 at Ser67 and/or Thr75 attenuates stimulatory effects of protein kinase A signaling in cardiac myocytes. AB - The depressed function of failing hearts has been partially attributed to increased protein phosphatase-1 through its impaired regulation by inhibitor-1. Phosphorylation of inhibitor-1 at Thr35 by PKA results in potent inhibition of protein phosphatase-1 activity, while phosphorylation at Ser67 or Thr75 by PKC attenuates the inhibitory activity. To examine the functional role of dual-site (Ser67, Thr75) phosphorylation of inhibitor-1 by PKC, the constitutively phosphorylated Ser67 (S67D) and/or Thr75 (T75D) human inhibitor-1 forms were expressed in adult cardiomyocytes. Expression of either single or double phosphorylated inhibitor-1 was associated with similar decreases in cardiac contractility, indicating that maximal inhibition can be elicited by each of these sites alone and that their inhibitory effects are not additive. Notably, activation of the cAMP pathway could only partially reverse the depressed contractile parameters. Accordingly, protein phosphatase-1 activity remained elevated, phosphorylation of phospholamban at Ser16 was decreased, and the EC(50) values of the sarcoplasmic reticulum calcium transport system were higher compared with controls. Thus phosphorylation of Ser67 and/or Thr75 in inhibitor-1 may mitigate the stimulatory effects of the cAMP pathway, resulting in compromised cardiac function. PMID- 17416611 TI - Molecular basis for PP2A regulatory subunit B56alpha targeting in cardiomyocytes. AB - Protein phosphatase 2A (PP2A) is a multifunctional protein phosphatase with critical roles in excitable cell signaling. In the heart, PP2A function is linked with modulation of beta-adrenergic signaling and has been suggested to regulate key ion channels and transporters including Na/Ca exchanger, ryanodine receptor, inositol 1,4,5-trisphosphate receptor, and Na/K ATPase. Although many of the functional roles and molecular targets for PP2A in heart are known, little is established regarding the cellular pathways that localize specific PP2A isoform activities to subcellular sites. We report that the PP2A regulatory subunit B56alpha is an in vivo binding partner for ankyrin-B, an adapter protein required for normal subcellular localization of the Na/Ca exchanger, Na/K ATPase, and inositol 1,4,5-trisphosphate receptor. Ankyrin-B and B56alpha are colocalized and coimmunoprecipitate in primary cardiomyocytes. Using multiple strategies, we identified the structural requirements on B56alpha for ankyrin-B association as a 13 residue motif in the B56alpha COOH terminus not present in other B56 family polypeptides. Finally, we report that reduced ankyrin-B expression in primary ankyrin-B(+/-) cardiomyocytes results in disorganized distribution of B56alpha that can be rescued by exogenous expression of ankyrin-B. These new data implicate ankyrin-B as a critical targeting component for PP2A in heart and identify a new class of signaling proteins targeted by ankyrin polypeptides. PMID- 17416612 TI - Bending dynamics of fluctuating biopolymers probed by automated high-resolution filament tracking. AB - Microscope images of fluctuating biopolymers contain a wealth of information about their underlying mechanics and dynamics. However, successful extraction of this information requires precise localization of filament position and shape from thousands of noisy images. Here, we present careful measurements of the bending dynamics of filamentous (F-)actin and microtubules at thermal equilibrium with high spatial and temporal resolution using a new, simple but robust, automated image analysis algorithm with subpixel accuracy. We find that slender actin filaments have a persistence length of approximately 17 microm, and display a q(-4)-dependent relaxation spectrum, as expected from viscous drag. Microtubules have a persistence length of several millimeters; interestingly, there is a small correlation between total microtubule length and rigidity, with shorter filaments appearing softer. However, we show that this correlation can arise, in principle, from intrinsic measurement noise that must be carefully considered. The dynamic behavior of the bending of microtubules also appears more complex than that of F-actin, reflecting their higher-order structure. These results emphasize both the power and limitations of light microscopy techniques for studying the mechanics and dynamics of biopolymers. PMID- 17416613 TI - Scattering of exciting light by live cells in fluorescence confocal imaging: phototoxic effects and relevance for FRAP studies. AB - As exciting light in a scanning confocal microscope encounters a cell and its subcellular components, it is refracted and scattered. A question arises as to what proportion of the exciting light is scattered by subcellular structures and whether cells in the vicinity of the imaged area, i.e., cells that are not directly illuminated by the laser beam, can be affected by either an exposure to scattered light and ensuing phototoxic reactions, or by the products of photoactivated reactions diffusing out of the directly illuminated area. We have designed a technique, which allows us to detect subtle cell photodamage and estimate the extent and range of phototoxic effects inflicted by interaction between scattered exciting light and fluorescent probes in the vicinity of the illuminated area. The technique is based on detecting an increased influx of acridine orange into photodamaged cells, which is manifested by a change of color. We demonstrate that phototoxic effects can be exerted not only on the illuminated cell, but also on fluorescently labeled neighboring cells. The damage inflicted on neighbors is due to exposure to light scattered by the imaged (i.e., directly illuminated) cell, but not phototoxic products diffusing out of the directly illuminated area. When light encounters a cell nucleus, scattering is so intense that photodamage can be inflicted even on fluorescently labeled cells located within a radius of approximately 90 microm, i.e., several cell diameters away. This range of scattering is comparable with that caused by the glass bead resting on a coverslip (up to 120 microm). The intense scattering of exciting light imposes limits on FRAP, FLIP, and other techniques employing high intensity laser beams. PMID- 17416614 TI - A freeze-fracture transmission electron microscopy and small angle x-ray diffraction study of the effects of albumin, serum, and polymers on clinical lung surfactant microstructure. AB - Freeze-fracture transmission electron microscopy shows significant differences in the bilayer organization and fraction of water within the bilayer aggregates of clinical lung surfactants, which increases from Survanta to Curosurf to Infasurf. Albumin and serum inactivate all three clinical surfactants in vitro; addition of the nonionic polymers polyethylene glycol, dextran, or hyaluronic acid also reduces inactivation in all three. Freeze-fracture transmission electron microscopy shows that polyethylene glycol, hyaluronic acid, and albumin do not adsorb to the surfactant aggregates, nor do these macromolecules penetrate the interior water compartments of the surfactant aggregates. This results in an osmotic pressure difference that dehydrates the bilayer aggregates, causing a decrease in the bilayer spacing as shown by small angle x-ray scattering and an increase in the ordering of the bilayers as shown by freeze-fracture electron microscopy. Small angle x-ray diffraction shows that the relationship between the bilayer spacing and the imposed osmotic pressure for Curosurf is a screened electrostatic interaction with a Debye length consistent with the ionic strength of the solution. The variation in surface tension due to surfactant adsorption measured by the pulsating bubble method shows that the extent of surfactant aggregate reorganization does not correlate with the maximum or minimum surface tension achieved with or without serum in the subphase. Albumin, polymers, and their mixtures alter the surfactant aggregate microstructure in the same manner; hence, neither inhibition reversal due to added polymer nor inactivation due to albumin is caused by alterations in surfactant microstructure. PMID- 17416615 TI - Nonequivalence of membrane voltage and ion-gradient as driving forces for the bacterial flagellar motor at low load. AB - Many bacterial species swim using flagella. The flagellar motor couples ion flow across the cytoplasmic membrane to rotation. Ion flow is driven by both a membrane potential (V(m)) and a transmembrane concentration gradient. To investigate their relation to bacterial flagellar motor function we developed a fluorescence technique to measure V(m) in single cells, using the dye tetramethyl rhodamine methyl ester. We used a convolution model to determine the relationship between fluorescence intensity in images of cells and intracellular dye concentration, and calculated V(m) using the ratio of intracellular/extracellular dye concentration. We found V(m) = -140 +/- 14 mV in Escherichia coli at external pH 7.0 (pH(ex)), decreasing to -85 +/- 10 mV at pH(ex) 5.0. We also estimated the sodium-motive force (SMF) by combining single-cell measurements of V(m) and intracellular sodium concentration. We were able to vary the SMF between -187 +/- 15 mV and -53 +/- 15 mV by varying pH(ex) in the range 7.0-5.0 and extracellular sodium concentration in the range 1-85 mM. Rotation rates for 0.35-microm- and 1 microm-diameter beads attached to Na(+)-driven chimeric flagellar motors varied linearly with V(m). For the larger beads, the two components of the SMF were equivalent, whereas for smaller beads at a given SMF, the speed increased with sodium gradient and external sodium concentration. PMID- 17416616 TI - GFP-mut2 proteins in trehalose-water matrixes: spatially heterogeneous protein water-sugar structures. AB - We report investigations on the properties of nanoenvironments around single-GFP mut2 proteins in trehalose-water matrixes. Single-GFPmut2 molecules embedded in thin trehalose-water films were characterized in terms of their fluorescence brightness, bleaching dynamics, excited state lifetime, and fluorescence polarization. For each property, sets of approximately 100-150 single molecules have been investigated as a function of trehalose content and hydration. Three distinct and interconverting families of proteins have been found which differ widely in terms of bleaching dynamics, brightness, and fluorescence polarization, whose relative populations sizably depend on sample hydration. The reported results evidence the simultaneous presence of different protein-trehalose-water nanostructures whose rigidity increases by lowering the sample hydration. Such spatial inhomogeneity is in line with the well-known heterogeneous dynamics in supercooled fluids and in nonsolid carbohydrate glasses and gives a pictorial representation of the sharp, sudden reorganization of the above structures after uptake <==>release of water molecules. PMID- 17416617 TI - Molecular dynamics simulations of indolicidin association with model lipid bilayers. AB - Identifying the mechanisms responsible for the interaction of peptides with cell membranes is critical to the design of new antimicrobial peptides and membrane transporters. We report here the results of a computational simulation of the interaction of the 13-residue peptide indolicidin with single-phase lipid bilayers of dioleoylphosphatidylcholine, distearoylphosphatidylcholine, dioleoylphosphatidylglycerol, and distearoylphosphatidylglycerol. Ensemble analysis of the membrane-bound peptide revealed that, in contrast to the extended, linear backbone structure reported for indolicidin in sodium dodecyl sulphate detergent micelles, the peptide adopts a boat-shaped conformation in both phosphatidylglycerol and phosphatidylcholine lipid bilayers, similar to that reported for dodecylphosphocholine micelles. In agreement with fluorescence and NMR experiments, simulations confirmed that the peptide localizes in the membrane interface, with the distance between phosphate headgroups of each leaflet being reduced in the presence of indolicidin. These data, along with a concomitant decrease in lipid order parameters for the upper-tail region, suggest that indolicidin binding results in membrane thinning, consistent with recent in situ atomic force microscopy studies. PMID- 17416618 TI - Protein hydrophobic collapse and early folding steps observed in a microfluidic mixer. AB - We demonstrate that the sub-millisecond protein folding process referred to as "collapse" actually consists of at least two separate processes. We observe the UV fluorescence spectrum from naturally occurring tryptophans in three well studied proteins, cytochrome c, apomyoglobin, and lysozyme, as a function of time in a microfluidic mixer with a dead time of approximately 20 mus. Single value decomposition of the time-dependent spectra reveal two separate processes: 1), a spectral shift which occurs within the mixing time; and 2), a fluorescence decay occurring between approximately 100 and 300 micros. We attribute the first process to hydrophobic collapse and the second process to the formation of the first native tertiary contacts. PMID- 17416619 TI - Systematic colocalization errors between acridine orange and EGFP in astrocyte vesicular organelles. AB - Dual-color imaging of acridine orange (AO) and EGFP fused to a vesicular glutamate transporter or the vesicle-associated membrane proteins 2 or 3 has been used to visualize a supposedly well-defined subpopulation of glutamatergic astrocytic secretory vesicles undergoing regulated exocytosis. However, AO metachromasy results in the concomitant emission of green and red fluorescence from AO-stained tissue. Therefore, the question arises whether AO and EGFP fluorescence can be distinguished reliably. We used evanescent-field imaging with spectral fluorescence detection as well as fluorescence lifetime imaging microscopy to demonstrate that green fluorescent AO monomers inevitably coexist with red fluorescing AO dimers, at the level of single astroglial vesicles. The green monomer emission spectrally overlaps with that of EGFP and produces a false apparent colocalization on dual-color images. On fluorophore abundance maps calculated from spectrally resolved and unmixed single-vesicle spectral image stacks, EGFP is obscured by the strong green monomer fluorescence, precluding the detection of EGFP. Hence, extreme caution is required when deriving quantitative colocalization information from images of dim fluorescing EGFP-tagged organelles colabeled with bright and broadly emitting dyes like AO. We finally introduce FM4 64/EGFP dual-color imaging as a remedy for imaging a distinct population of astroglial fusion-competent secretory vesicles. PMID- 17416620 TI - Substrate chemistry-dependent conformations of single laminin molecules on polymer surfaces are revealed by the phase signal of atomic force microscopy. AB - The conformation of single laminin molecules adsorbed on synthetic substrates is directly observed making use of the phase magnitude in tapping mode atomic force microscopy (AFM). With AFM, it is not possible to differentiate the proteins on the substrate if use is made of the height signal, since the roughness of the material becomes of the same order of magnitude as the adsorbed protein, typically 10 nm height. This work shows how AFM can be exploited to reveal protein conformation on polymer materials. Different laminin morphologies are observed on a series of different copolymers based on ethyl acrylate and hydroxyethyl acrylate as a function of the surface density of -OH groups: from globular to completely extended morphologies of the protein molecules are obtained, and the onset of laminin network formation on some substrates can be clearly identified. The results stress the importance of the underlying synthetic substrate's surface chemistry for the biofunctional conformation of adsorbed proteins. PMID- 17416621 TI - Molecular mechanism of the Zn2+-induced folding of the distal CCHC finger motif of the HIV-1 nucleocapsid protein. AB - HIV-1 nucleocapsid protein, NCp7, contains two highly conserved CCHC zinc fingers. Binding of Zn(2+) drives NCp7 from an unfolded to a highly folded structure that is critical for its functions. Using the intrinsic fluorescence of Trp(37), we investigated, by the stopped-flow technique, the folding of NCp7 distal finger through the pH dependence of its Zn(2+) association and dissociation kinetics. Zn(2+) binding was found to involve four different paths associated with the four deprotonated states of the finger. Each binding path involves the rapid formation of an intermediate complex that is subsequently rearranged and stabilized in a rate-limiting step. The equilibrium and kinetic rate constants of the full Zn(2+)-binding process have been determined. At neutral pH, the preferential pathway for the Zn(2+)-driven folding implies Zn(2+) binding to the deprotonated Cys(36) and His(44) residues, in the bidentate state of the finger. The resulting intermediate is then converted with a rate constant of 500 s(-1) into a more suitably folded form, probably through a rearrangement of the peptide backbone around Zn(2+) to optimize the binding geometry. This form then rapidly leads to the final native complex, through deprotonation of Cys(39) and Cys(49) residues and intramolecular substitution of coordinated water molecules. Zn(2+) dissociation is also characterized by a multistep process and occurs fastest via the deprotonated Zn(2+)-bound bidentate state with a rate constant of 3 s(-1). Due to their critical role in folding, the intermediates identified for the first time in this study may constitute potential targets for HIV therapy. PMID- 17416622 TI - Estimation of ion channel kinetics from fluctuations of macroscopic currents. AB - For single channel recordings, the maximum likelihood estimation (MLE) of kinetic rates and conductance is well established. A direct extrapolation of this method to macroscopic currents is computationally prohibitive: it scales as a power of the number of channels. An approximated MLE that ignored the local time correlation of the data has been shown to provide estimates of the kinetic parameters. In this article, an improved approximated MLE that takes into account the local time correlation is proposed. This method estimates the channel kinetics using both the time course and the random fluctuations of the macroscopic current generated by a homogeneous population of ion channels under white noise. It allows arbitrary kinetic models and stimulation protocols. The application of the proposed algorithm to simulated data from a simple three-state model on nonstationary conditions showed reliable estimates of all the kinetic constants, the conductance and the number of channels, and reliable values for the standard error of those estimates. Compared to the previous approximated MLE, it reduces by a factor of 10 the amount of data needed to secure a given accuracy and it can even determine the kinetic rates in macroscopic stationary conditions. PMID- 17416623 TI - Optimal specificity and function for flexible biomolecular recognition. AB - Biomolecular associations often accompanied by large conformational changes, sometimes folding and unfolding. By exploring an exactly solvable model, we constructed the free energy landscape and established a general framework for studying the biomolecular flexible binding process. We derived an optimal criterion for the specificity and function for flexible biomolecular binding where the binding and conformational folding are coupled. PMID- 17416624 TI - Membrane-binding/modification model of signaling protein activation and analysis of its control by cell morphology. AB - A mechanism for cell shape control of intracellular signal transduction, whereby the average concentration of activated proteins in the cytosol increases as the height of the cell decreases, has been described recently. An important modification of this analysis is offered, recognizing that signaling proteins are not only activated at the plasma membrane but must first form complexes with signaling molecules that reside there, such as receptors and lipids. With these more realistic boundary conditions, it is shown that the region of parameter space where cell shape amplifies the average cytosolic activity is greatly expanded. Moreover, this model allows for amplification of the activated protein bound at the membrane, which is considered more relevant for certain, spatially driven signaling processes in cell migration. PMID- 17416625 TI - Different effects of lipid chain length on the two sides of a membrane and the lipid annulus of MscL. AB - Quenching of the fluorescence of Trp residues in a membrane protein by lipids with bromine-containing fatty acyl chains provides a powerful technique for measuring lipid-protein binding constants. Single Trp residues have been placed on the periplasmic and cytoplasmic sides of the mechanosensitive channel of large conductance MscL from Mycobacterium tuberculosis to measure, separately, lipid binding constants on the two faces of MscL. The chain-length dependence of lipid binding was found to be different on the two sides of MscL, the chain-length dependence being more marked on the cytoplasmic than on the periplasmic side. To determine if lipid binding constants are affected by the properties of the lipid molecules not in direct contact with MscL (the bulk lipid), the amount of bulk lipid present in the system was varied. The binding constant of the short-chain phospholipid didodecylphosphatidylcholine was found to be independent of the molar ratio of lipid/MscL pentamer over the range 500:1-50:1, suggesting that lipid binding constants are determined largely by the properties of the lipid molecules interacting directly with MscL. These results point to a model in which lipid molecules located on the transmembrane surface of a membrane protein (the annular lipid molecules), by playing a dominant role in the interaction between a membrane protein and the surrounding lipid bilayer, could effectively buffer the membrane protein from changes in the properties of the bulk lipid bilayer. PMID- 17416626 TI - Single-molecule spectroscopic characterization of light-harvesting 2 complexes reconstituted into model membranes. AB - The spectroscopic properties of the light-harvesting 2 complexes (LH2) from the purple bacterium Rhodopseudomonas acidophila (strain 10050) in detergent micelles and reconstituted into lipid membranes have been studied by single-molecule spectroscopy. When LH2 complexes are solubilized from their host biological membranes by nondenaturing detergents, such as LDAO, there is a small 2-nm spectral shift of the B850 absorption band in the ensemble spectrum. This is reversed when the LH2 complexes are put back into phospholipid vesicles, i.e., into a more native-like environment. The spectroscopic properties on the single molecule level of the detergent-solubilized LH2 complexes were compared with those reconstituted into the lipid membranes to see if their detailed spectroscopic behavior was influenced by these small changes in the position of the B850 absorption band. A detailed analysis of the low-temperature single molecule fluorescence-excitation spectra of the LH2 complexes in these two different conditions showed no significant differences. In particular, the distribution of the spectral splitting between the circular k = +/-1 exciton states of the B850 absorption band and the distribution of the mutual angle between the k = +/-1 exciton states are identical in both cases. It can be concluded, therefore, that the LH2 complexes from Rps. acidophila are equally stable when solubilized in detergent micelles as they are when membrane reconstituted. Moreover, when they are solubilized in a suitable detergent and spin coated onto a surface for the single-molecule experiments they do not display any more structural disorder than when in a phospholipid membrane. PMID- 17416627 TI - A fiber-based ratiometric optical cardiac mapping channel using a diffraction grating and split detector. AB - Optical fiber-based mapping systems are used to record the cardiac action potential (AP) throughout the myocardium. The optical AP contains a contraction induced motion artifact (MA), which makes it difficult to accurately measure the action potential duration (APD). MA is removed by preventing contraction with electrical-mechanical uncoupling drugs, such as 2,3-butanedione monoxime (BDM). We designed a novel fiber-based ratiometric optical channel using a blue light emitting diode, a diffraction grating, and a split photodetector that can accurately measure the cardiac AP without the need for BDM. The channel was designed based on simulations using the optical design software ZEMAX. The channel has an electrical bandwidth of 150 Hz and an root mean-square dark noise of 742 muV. The channel successfully recorded the cardiac AP from the wall of five rabbit heart preparations without the use of BDM. After 20-point median filtering, the mean signal/noise ratio was 25.3 V/V. The APD measured from the base of a rabbit heart was 134 +/- 8.4 ms, compared to 137.6 +/- 3.3 ms from simultaneous microelectrode recordings. This difference was not statistically significant (p-value = 0.3). The quantity of MA removed was also measured using the motion ratio. The reduction in MA was significant (p-value = 0.0001). This fiber-based system is the first of its kind to enable optical APD measurements in the beating heart wall without the use of BDM. PMID- 17416628 TI - Changing the charge distribution of beta-helical-based nanostructures can provide the conditions for charge transfer. AB - In this work we present a computational approach to the design of nanostructures made of structural motifs taken from left-handed beta-helical proteins. Previously, we suggested a structural model based on the self-assembly of motifs taken from Escherichia coli galactoside acetyltransferase (Protein Data Bank 1krr, chain A, residues 131-165, denoted krr1), which produced a very stable nanotube in molecular dynamics simulations. Here we modify this model by changing the charge distribution in the inner core of the system and testing the effect of this change on the structural arrangement of the construct. Our results demonstrate that it is possible to generate the proper conditions for charge transfer inside nanotubes based on assemblies of krr1 segment. The electronic transfer would be achieved by introducing different histidine ionization states in selected positions of the internal core of the construct, in addition to specific mutations with charged amino acids that altogether will allow the formation of coherent networks of aromatic ring stacking, salt-bridges, and hydrogen bonds. PMID- 17416629 TI - Pore formation by a Bax-derived peptide: effect on the line tension of the membrane probed by AFM. AB - Bax is a critical regulator of physiological cell death that increases the permeability of the outer mitochondrial membrane and facilitates the release of the so-called apoptotic factors during apoptosis. The molecular mechanism of action is unknown, but it probably involves the formation of partially lipidic pores induced by Bax. To investigate the interaction of Bax with lipid membranes and the physical changes underlying the formation of Bax pores, we used an active peptide derived from helix 5 of this protein (Bax-alpha5) that is able to induce Bax-like pores in lipid bilayers. We report the decrease of line tension due to peptide binding both at the domain interface in phase-separated lipid bilayers and at the pore edge in atomic force microscopy film-rupture experiments. Such a decrease in line tension may be a general strategy of pore-forming peptides and proteins, as it affects the energetics of the pore and stabilizes the open state. PMID- 17416630 TI - Stochastic signal processing and transduction in chemotactic response of eukaryotic cells. AB - Single-molecule imaging analysis of chemotactic response in eukaryotic cells has revealed a stochastic nature in the input signals and the signal transduction processes. This leads to a fundamental question about the signaling processes: how does the signaling system operate under stochastic fluctuations or noise? Here, we report a stochastic model of chemotactic signaling in which noise and signal propagation along the transmembrane signaling pathway by chemoattractant receptors can be analyzed quantitatively. The results obtained from this analysis reveal that the second-messenger-production reactions by the receptors generate noisy signals that contain intrinsic noise inherently generated at this reaction and extrinsic noise propagated from the ligand-receptor binding. Such intrinsic and extrinsic noise limits the directional sensing ability of chemotactic cells, which may explain the dependence of chemotactic accuracy on chemical gradients that has been observed experimentally. Our analysis also reveals regulatory mechanisms for signal improvement in the stochastically operating signaling system by analyzing how the SNR of chemotactic signals can be improved on or deteriorated by the stochastic properties of receptors and second-messenger molecules. Theoretical consideration of noisy signal transduction by chemotactic signaling systems can further be applied to signaling systems in general. PMID- 17416631 TI - Probing the nanoscale viscoelasticity of intracellular fluids in living cells. AB - We have used fluorescence correlation spectroscopy to determine the anomalous diffusion properties of fluorescently tagged gold beads in the cytoplasm and the nucleus of living cells. From the extracted mean-square displacement v(tau) approximately tau(alpha), we have determined the complex shear modulus G(omega) approximately omega(alpha) for both compartments. Without treatment, all tested cell lines showed a strong viscoelastic behavior of the cytoplasm and the nucleoplasm, highlighting the crowdedness of these intracellular fluids. We also found a similar viscoelastic response in frog egg extract, which tended toward a solely viscous behavior upon dilution. When cells were osmotically stressed, the diffusion became less anomalous and the viscoelastic response changed. In particular, the anomality changed from alpha approximately 0.55 to alpha approximately 0.66, which indicates that the Zimm model for polymer solutions under varying solvent conditions is a good empirical description of the material properties of the cytoplasm and the nucleoplasm. Since osmotic stress may eventually trigger cell death, we propose, on the basis of our observations, that intracellular fluids are maintained in a state similar to crowded polymer solutions under good solvent conditions to keep the cell viable. PMID- 17416632 TI - Enumeration of oligomerization states of membrane proteins in living cells by homo-FRET spectroscopy and microscopy: theory and application. AB - Protein-protein interactions play a pivotal role in biological signaling networks. It is highly desirable to perform experiments that can directly assess the oligomerization state and degree of oligomerization of biological macromolecules in their native environment. Homo-FRET depends on the inverse sixth power of separation between interacting like fluorophores on the nanometer scale and is therefore sensitive to protein oligomerization. Homo-FRET is normally detected by steady-state or time-resolved fluorescence anisotropy measurements. Here we show by theory and simulation that an examination of the extent of homotransfer as measured by steady-state fluorescence anisotropy as a function of fluorophore labeling (or photodepletion) gives valuable information on the oligomerization state of self-associating proteins. We examine random distributions of monomers, dilute solutions of oligomers, and concentrated solutions of oligomers. The theory is applied to literature data on band 3 protein dimers in membranes, GPI-linked protein trimers in "rafts," and clustered GFP-tagged epidermal growth factor receptors in cell membranes to illustrate the general utility and applicability of our analytical approach. PMID- 17416633 TI - RNA-editing-associated protein 1 null mutant reveals link to mitochondrial RNA stability. AB - In trypanosomatids, uridylate residues are post-transcriptionally added to or deleted from pre-mRNAs during the complex process of RNA editing. Editing is carried out exclusively in the mitochondrion of these parasites and involves numerous proteins assembled into protein and ribonucleoprotein complexes. Previously we identified RNA-editing-associated protein -1 (REAP-1), an RNA binding protein found in the mitochondrion of Trypanosoma brucei. REAP-1 was shown to specifically recognize and bind to pre-mRNAs that require editing and was proposed to act as a recruitment factor to deliver pre-mRNAs to editing complexes. To help define the role of REAP-1, we have now constructed REAP-1 null mutants. We show that the null mutants, although viable, have a significant growth defect. RNA levels within the mitochondrion were evaluated using reverse transcriptase real-time PCR. Surprisingly, the results show that mitochondrial RNA levels are increased, regardless of the editing status of the RNA. All RNA tested, whether unedited, edited, or never edited were increased in the mutant cell line relative to wild-type levels. This study provides the first evidence for a role of REAP-1 in RNA metabolism. PMID- 17416634 TI - Structures of tRNAs with an expanded anticodon loop in the decoding center of the 30S ribosomal subunit. AB - During translation, some +1 frameshift mRNA sites are decoded by frameshift suppressor tRNAs that contain an extra base in their anticodon loops. Similarly engineered tRNAs have been used to insert nonnatural amino acids into proteins. Here, we report crystal structures of two anticodon stem-loops (ASLs) from tRNAs known to facilitate +1 frameshifting bound to the 30S ribosomal subunit with their cognate mRNAs. ASL(CCCG) and ASL(ACCC) (5'-3' nomenclature) form unpredicted anticodon-codon interactions where the anticodon base 34 at the wobble position contacts either the fourth codon base or the third and fourth codon bases. In addition, we report the structure of ASL(ACGA) bound to the 30S ribosomal subunit with its cognate mRNA. The tRNA containing this ASL was previously shown to be unable to facilitate +1 frameshifting in competition with normal tRNAs (Hohsaka et al. 2001), and interestingly, it displays a normal anticodon-codon interaction. These structures show that the expanded anticodon loop of +1 frameshift promoting tRNAs are flexible enough to adopt conformations that allow three bases of the anticodon to span four bases of the mRNA. Therefore it appears that normal triplet pairing is not an absolute constraint of the decoding center. PMID- 17416635 TI - Induction of drug resistance and transformation in human cancer cells by the noncoding RNA CUDR. AB - Refractory to apoptosis induced by anticancer drugs is one of the major causes of drug resistance in human cancers. The involvement of noncoding RNA (ncRNA) in cancer cell drug resistance has not yet been reported. By using the technique of RT-PCR-based differential display, a novel gene, cancer up-regulated drug resistant (CUDR) gene, was found to be overexpressed in a doxorubicin-resistant subline of human squamous carcinoma A431 and A10A cells, which were also more resistant to drug-induced apoptosis. The full-length CUDR mRNA transcript is approximately 2.2 kb as detected by Northern blot analysis and has no sequence homology with other genes identified so far. Interestingly, no distinct open reading frame was found throughout the CUDR cDNA sequence, and no recombinant protein was detected from in vitro translation or from a protein lysate of human cancer cells after CUDR transfection. Therefore, CUDR is likely to exert its function as a noncoding RNA. Stable transfection with the CUDR gene was found to induce resistance to doxorubicin and etoposide as well as drug-induced apoptosis in A431 cells. By Western blot analysis, down-regulations of caspase 3 were observed in CUDR transfectants. On the other hand, overexpression of CUDR promoted anchorage-independent growth in A431 cells. Results from the present study suggest that CUDR may likely regulate the drug sensitivity and promote cellular transformation at least through caspase 3-dependent apoptosis. PMID- 17416636 TI - Induction of isoforms of tetrapyrrole biosynthetic enzymes, AtHEMA2 and AtFC1, under stress conditions and their physiological functions in Arabidopsis. AB - In the tetrapyrrole biosynthetic pathway, isoforms of glutamyl-tRNA reductase (HEMA2) and ferrochelatase1 (FC1) are mainly expressed in nonphotosynthetic tissues. Here, using promoter-beta-glucuronidase constructs, we showed that the expressions of Arabidopsis (Arabidopsis thaliana) HEMA2 (AtHEMA2) and FC1 (AtFC1) were induced in photosynthetic tissues by oxidative stresses such as wounding. Transcript levels and beta-glucronidase activity were rapidly induced within 30 min, specifically in the wound area in a jasmonate-independent manner. Transcriptome analysis of wound-specific early inducible genes showed that AtHEMA2 and AtFC1 were coinduced with hemoproteins outside plastids, which are related to defense responses. Ozone fumigation or reagents generating reactive oxygen species induced the expression of both genes in photosynthetic tissues, suggesting that reactive oxygen species is involved in the induction. Since cycloheximide or puromycin induced the expression of both genes, inhibition of cytosolic protein synthesis is involved in the induction of these genes in photosynthetic tissues. The physiological functions of AtHEMA2 and AtFC1 were investigated using insertional knockout mutants of each gene. Heme contents of the roots of both mutants were about half of that of the respective wild types. In wild-type plants, heme contents were increased by ozone exposure. In both mutants, reduction of the ozone-induced increase in heme content was observed. These results suggest the existence of the tetrapyrrole biosynthetic pathway controlled by AtHEMA2 and AtFC1, which normally functions for heme biosynthesis in nonphotosynthetic tissues, but is induced in photosynthetic tissues under oxidative conditions to supply heme for defensive hemoproteins outside plastids. PMID- 17416637 TI - RNAi silencing of genes for elicitation or biosynthesis of 5-deoxyisoflavonoids suppresses race-specific resistance and hypersensitive cell death in Phytophthora sojae infected tissues. AB - Isoflavonoids are thought to play an important role in soybean (Glycine max) resistance to Phytophthora sojae. This was addressed by silencing two genes for their biosynthesis and a third gene controlling their elicitation. Silencing of genes for isoflavone synthase (IFS) or chalcone reductase (CHR) was achieved in soybean roots through an Agrobacterium rhizogenes-mediated RNAi approach. Effectiveness of silencing was followed both by quantitative reverse transcriptase-polymerase chain reaction and high-performance liquid chromatography analyses. Silencing either IFS or CHR led to a breakdown of Rps mediated resistance to race 1 of P. sojae in 'W79' (Rps 1c) or 'W82' (Rps 1k) soybean. Loss of resistance was accompanied by suppression of hypersensitive (HR) cell death in both cultivars and suppression of cell death-associated activation of hydrogen peroxide and peroxidase. The various results suggest that the 5 deoxyisoflavonoids play a critical role in the establishment of cell death and race-specific resistance. The P. sojae cell wall glucan elicitor, a potent elicitor of 5-deoxyisoflavonoids, triggered a cell death response in roots that was also suppressed by silencing either CHR or IFS. Furthermore, silencing of the elicitor-releasing endoglucanase (PR-2) led to a loss of HR cell death and race specific resistance to P. sojae and also to a loss of isoflavone and cell death responses to cell wall glucan elicitor. Taken together, these results suggest that in situ release of active fragments from a general resistance elicitor (pathogen-associated molecular pattern) is necessary for HR cell death in soybean roots carrying resistance genes at the Rps 1 locus, and that this cell death response is mediated through accumulations of the 5-deoxyisoflavones. PMID- 17416638 TI - Sebacina vermifera promotes the growth and fitness of Nicotiana attenuata by inhibiting ethylene signaling. AB - Sebacina vermifera, a growth-promoting endophytic fungus, significantly increases Nicotiana attenuata's growth but impairs both its herbivore resistance and its accumulation of the costly, jasmonic acid (JA)-regulated defense protein, trypsin proteinase inhibitor (TPI). To determine if the fungi's growth-promoting effects can be attributed to lower TPI-related defense costs, we inoculated transformed N. attenuata plants silenced in their ability to synthesize JA, JA-isoleucine, and TPI by antisense (lipoxygenase 3 [as-lox3] and Thr deaminase [as-td]) and inverted repeat (ir-tpi) expression, and found that inoculation promoted plant growth as in untransformed wild-type plants. Moreover, herbivore-elicited increases in JA and JA-isoleucine concentrations did not differ between inoculated and uninoculated wild-type plants. However, inoculation significantly reduced the morphological effect of 1-aminocyclopropane-1-carboxylic acid on wild type seedlings in a triple response assay, suggesting that ethylene signaling was impaired. Furthermore, S. vermifera failed to promote the growth of N. attenuata plants transformed to silence ethylene production (1-aminocyclopropane-1 carboxylic acid oxidase [ir-aco]). Inoculating wild-type plants with S. vermifera decreased the ethylene burst elicited by applying Manduca sexta oral secretions to mechanical wounds. Accordingly, oral secretion-elicited transcript levels of the ethylene synthesis genes NaACS3, NaACO1, and NaACO3 in inoculated plants were significantly lower compared to these levels in uninoculated wild-type plants. Inoculation accelerated germination in wild-type seeds; however, uninoculated wild-type seeds germinated as rapidly as inoculated seeds in the presence of the ethylene scrubber KMnO(4). In contrast, neither inoculation nor KMnO(4) exposure influenced the germination of ir-aco seeds. We conclude that S. vermifera increases plant growth by impairing ethylene production independently of JA signaling and TPI production. PMID- 17416639 TI - REP27, a tetratricopeptide repeat nuclear-encoded and chloroplast-localized protein, functions in D1/32-kD reaction center protein turnover and photosystem II repair from photodamage. AB - The goal of this research is elucidation of the molecular mechanism for the unique photosystem II (PSII) damage and repair cycle in chloroplasts. A frequently occurring, irreversible photooxidative damage inhibits the PSII charge separation reaction and stops photosynthesis. The chloroplast PSII repair process rectifies this adverse effect by selectively removing and replacing the photoinactivated D1/32-kD reaction center protein (the chloroplast-encoded psbA gene product) from the massive (>1,000 kD) water-oxidizing and O2-evolving PSII holocomplex. DNA insertional mutagenesis in the model organism Chlamydomonas reinhardtii was applied for the isolation and characterization of rep27, a repair aberrant mutant. Gene cloning and biochemical analyses in this mutant resulted in the identification of REP27, a nuclear gene encoding a putative chloroplast targeted protein, which is specifically required for the completion of the D1 turnover process but is not essential for the de novo biogenesis and assembly of the PSII holocomplex in this model green alga. The REP27 protein contains two highly conserved tetratricopeptide repeats, postulated to facilitate the psbA mRNA cotranslational insertion of the nascent D1 protein in the existing PSII core template. Elucidation of the PSII repair mechanism may reveal the occurrence of hitherto unknown regulatory and catalytic reactions for the selective in situ replacement of specific proteins from within multiprotein complexes. PMID- 17416640 TI - Involvement of the histone acetyltransferase AtHAC1 in the regulation of flowering time via repression of FLOWERING LOCUS C in Arabidopsis. AB - Histone acetylation is an important posttranslational modification correlated with gene activation. In Arabidopsis (Arabidopsis thaliana), the histone acetyltransferase AtHAC1 is homologous to animal p300/CREB (cAMP-responsive element-binding protein)-binding proteins, which are the main histone acetyltransferases participating in many physiological processes, including proliferation, differentiation, and apoptosis. The functions of p300/CREB-binding proteins in animals are well characterized, whereas little is known about the roles of AtHAC1 in developmental control in Arabidopsis. Lesions in AtHAC1 caused pleiotropic developmental defects, including delayed flowering, a shortened primary root, and partially reduced fertility. Analysis of the molecular basis of late flowering in hac1 mutants showed that the hac1 plants respond normally to day length, gibberellic acid treatment, and vernalization. Furthermore, the expression level of the flowering repressor FLOWERING LOCUS C (FLC) is increased in hac1 mutants, indicating that the late-flowering phenotype of hac1 mutants is mediated by FLC. Since histone acetylation is usually associated with the activation of gene expression, histone modifications of FLC chromatin are not affected by mutations in HAC1 and expression levels of all known autonomous pathway genes are unchanged in hac1 plants, we propose that HAC1 affects flowering time by epigenetic modification of factors upstream of FLC. PMID- 17416641 TI - Regulation of arbuscular mycorrhization by carbon. The symbiotic interaction cannot be improved by increased carbon availability accomplished by root specifically enhanced invertase activity. AB - The mutualistic interaction in arbuscular mycorrhiza (AM) is characterized by an exchange of mineral nutrients and carbon. The major benefit of AM, which is the supply of phosphate to the plant, and the stimulation of mycorrhization by low phosphate fertilization has been well studied. However, less is known about the regulatory function of carbon availability on AM formation. Here the effect of enhanced levels of hexoses in the root, the main form of carbohydrate used by the fungus, on AM formation was analyzed. Modulation of the root carbohydrate status was performed by expressing genes encoding a yeast (Saccharomyces cerevisiae) derived invertase, which was directed to different subcellular locations. Using tobacco (Nicotiana tabacum) alcc::wINV plants, the yeast invertase was induced in the whole root system or in root parts. Despite increased hexose levels in these roots, we did not detect any effect on the colonization with Glomus intraradices analyzed by assessment of fungal structures and the level of fungus-specific palmitvaccenic acid, indicative for the fungal carbon supply, or the plant phosphate content. Roots of Medicago truncatula, transformed to express genes encoding an apoplast-, cytosol-, or vacuolar-located yeast-derived invertase, had increased hexose-to-sucrose ratios compared to beta-glucuronidase-transformed roots. However, transformations with the invertase genes did not affect mycorrhization. These data suggest the carbohydrate supply in AM cannot be improved by root-specifically increased hexose levels, implying that under normal conditions sufficient carbon is available in mycorrhizal roots. In contrast, tobacco rolC::ppa plants with defective phloem loading and tobacco pyk10::InvInh plants with decreased acid invertase activity in roots exhibited a diminished mycorrhization. PMID- 17416642 TI - The transiently generated nonphotochemical quenching of excitation energy in Arabidopsis leaves is modulated by zeaxanthin. AB - Upon the transition of dark-adapted plants to low light, the energy-dependent quenching (qE) of excitation energy is only transiently induced due to the only transient generation of the transthylakoid pH gradient. We investigated the transient qE (qE(TR)) in different Arabidopsis (Arabidopsis thaliana) mutants. In dark-adapted plants, qE(TR) was absent in the npq4 mutant (deficient in the PsbS protein) and the pgr1 mutant (restricted in lumen acidification). In comparison with wild-type plants, qE(TR) was reduced in the zeaxanthin (Zx)-deficient npq1 mutant and increased in the Zx-accumulating npq2 mutant. After preillumination of plants (to allow the synthesis of large amounts of Zx), the formation and relaxation of qE(TR) was accelerated in all plants (except for npq4) in comparison with the respective dark-adapted plants. The extent of qE(TR), however, was unchanged in npq1 and npq4, decreased in npq2, but increased in wild type and pgr1 plants. Even in presence of high levels of Zx, qE(TR) in pgr1 mutants was still lower than that in wild-type plants. In the presence of the uncoupler nigericin, qE(TR) was completely abolished in all genotypes. Thus, the transient qE(TR) shows essentially the same characteristics as the steady-state qE; it is strictly dependent on the PsbS protein and a low lumen pH, but the extent of qE(TR) is largely modulated by Zx. These results indicate that qE(TR) does not represent a different quenching mechanism in comparison with the steady state qE, but simply reflects the response of qE to the dynamics of the lumen pH during light activation of photosynthesis. PMID- 17416643 TI - Stability of plant defense proteins in the gut of insect herbivores. AB - Plant defense against insect herbivores is mediated in part by enzymes that impair digestive processes in the insect gut. Little is known about the evolutionary origins of these enzymes, their distribution in the plant kingdom, or the mechanisms by which they act in the protease-rich environment of the animal digestive tract. One example of such an enzyme is threonine (Thr) deaminase (TD), which in tomato (Solanum lycopersicum) serves a dual role in isoleucine (Ile) biosynthesis in planta and Thr degradation in the insect midgut. Here, we report that tomato uses different TD isozymes to perform these functions. Whereas the constitutively expressed TD1 has a housekeeping role in Ile biosynthesis, expression of TD2 in leaves is activated by the jasmonate signaling pathway in response to herbivore attack. Ingestion of tomato foliage by specialist (Manduca sexta) and generalist (Trichoplusia ni) insect herbivores triggered proteolytic removal of TD2's C-terminal regulatory domain, resulting in an enzyme that degrades Thr without being inhibited through feedback by Ile. This processed form (pTD2) of TD2 accumulated to high levels in the insect midgut and feces (frass). Purified pTD2 exhibited biochemical properties that are consistent with a postingestive role in defense. Shotgun proteomic analysis of frass from tomato-reared M. sexta identified pTD2 as one of the most abundant proteins in the excrement. Among the other tomato proteins identified were several jasmonate inducible proteins that have a known or proposed role in anti-insect defense. Subtilisin-like proteases and other pathogenesis-related proteins, as well as proteins of unknown function, were also cataloged. We conclude that proteomic analysis of frass from insect herbivores provides a robust experimental approach to identify hyperstable plant proteins that serve important roles in defense. PMID- 17416644 TI - Nitrogen fixation control under drought stress. Localized or systemic? AB - Legume-Rhizobium nitrogen fixation is dramatically affected under drought and other environmental constraints. However, it has yet to be established as to whether such regulation of nitrogen fixation is only exerted at the whole-plant level (e.g. by a systemic nitrogen feedback mechanism) or can also occur at a local nodule level. To address this question, nodulated pea (Pisum sativum) plants were grown in a split-root system, which allowed for half of the root system to be irrigated at field capacity, while the other half was water deprived, thus provoking changes in the nodule water potential. Nitrogen fixation only declined in the water-deprived, half-root system and this result was correlated with modifications in the activities of key nodule's enzymes such as sucrose synthase and isocitrate dehydrogenase and in nodular malate content. Furthermore, the decline in nodule water potential resulted in a cell redox imbalance. The results also indicate that systemic nitrogen feedback signaling was not operating in these water-stressed plants, since nitrogen fixation activity was maintained at control values in the watered half of the split-root plants. Thus, the use of a partially droughted split-root system provides evidence that nitrogen fixation activity under drought stress is mainly controlled at the local level rather than by a systemic nitrogen signal. PMID- 17416645 TI - Structural properties of periplasmic SodCI that correlate with virulence in Salmonella enterica serovar Typhimurium. AB - Salmonella enterica strains survive and propagate in macrophages by both circumventing and resisting the antibacterial effectors normally delivered to the phagosome. An important aspect of Salmonella resistance is the production of periplasmic superoxide dismutase to combat phagocytic superoxide. S. enterica serovar Typhimurium strain 14028 produces two periplasmic superoxide dismutases: SodCI and SodCII. Both enzymes are produced during infection, but only SodCI contributes to virulence in the animal. Although 60% identical to SodCII at the amino acid level with very similar enzymatic properties, SodCI is dimeric, protease resistant, and tethered within the periplasm via a noncovalent interaction. In contrast, SodCII is monomeric and protease sensitive and is released from the periplasm normally by osmotic shock. We have constructed an enzymatically active monomeric SodCI enzyme by site-directed mutagenesis. The resulting protein was released by osmotic shock and sensitive to protease and could not complement the loss of wild-type dimeric SodCI during infection. To distinguish which property is most critical during infection, we cloned and characterized related SodC proteins from a variety of bacteria. Brucella abortus SodC was monomeric and released by osmotic shock but was protease resistant and could complement SodCI in the animal. These data suggest that protease resistance is a critical property that allows SodCI to function in the harsh environment of the phagosome to combat phagocytic superoxide. We propose a model to account for the various properties of SodCI and how they contribute to bacterial survival in the phagosome. PMID- 17416646 TI - Predation by Bdellovibrio bacteriovorus HD100 requires type IV pili. AB - Early electron microscopy and more recent studies in our laboratory of Bdellovibrio bacteriovorus cells indicated the presence of narrow fibers at the nonflagellar pole of this unusual predatory bacterium. Analysis of the B. bacteriovorus HD100 genome showed a complete set of genes potentially encoding type IV pili and an incomplete gene set for Flp pili; therefore, the role of type IV pili in the predatory life cycle of B. bacteriovorus HD100 was investigated. Alignment of the predicted PilA protein with known type IV pilins showed the characteristic conserved N terminus common to type IVa pilins. The pilA gene, encoding the type IV pilus fiber protein, was insertionally inactivated in multiple Bdellovibrio replicate cultures, and the effect upon the expression of other pilus genes was monitored by reverse transcriptase PCR. Interruption of pilA in replicate isolates abolished Bdellovibrio predatory capability in liquid prey cultures and on immobilized yellow fluorescent protein-labeled prey, but the mutants could be cultured prey independently. Expression patterns of pil genes involved in the formation of type IV pili were profiled across the predatory life cycle from attack phase predatory Bdellovibrio throughout the intraperiplasmic bdelloplast stages to prey lysis and in prey-independent growth. Taken together, the data show that type IV pili play a critical role in Bdellovibrio predation. PMID- 17416647 TI - Flagellar motility is critical for Listeria monocytogenes biofilm formation. AB - The food-borne pathogen Listeria monocytogenes attaches to environmental surfaces and forms biofilms that can be a source of food contamination, yet little is known about the molecular mechanisms of its biofilm development. We observed that nonmotile mutants were defective in biofilm formation. To investigate how flagella might function during biofilm formation, we compared the wild type with flagellum-minus and paralyzed-flagellum mutants. Both nonmotile mutants were defective in biofilm development, presumably at an early stage, as they were also defective in attachment to glass during the first few hours of surface exposure. This attachment defect could be significantly overcome by providing exogenous movement toward the surface via centrifugation. However, this centrifugation did not restore mature biofilm formation. Our results indicate that it is flagellum mediated motility that is critical for both initial surface attachment and subsequent biofilm formation. Also, any role for L. monocytogenes flagella as adhesins on abiotic surfaces appears to be either minimal or motility dependent under the conditions we examined. PMID- 17416648 TI - Molecular analysis of genes in Nostoc punctiforme involved in pilus biogenesis and plant infection. AB - Hormogonia are the infective agents in many cyanobacterium-plant symbioses. Pilus like appendages are expressed on the hormogonium surface, and mutations in pil like genes altered surface piliation and reduced symbiotic competency. This is the first molecular evidence that pilus biogenesis in a filamentous cyanobacterium requires a type IV pilus system. PMID- 17416649 TI - SOS induction in a subpopulation of structural maintenance of chromosome (Smc) mutant cells in Bacillus subtilis. AB - The structural maintenance of chromosome (Smc) protein is highly conserved and involved in chromosome compaction, cohesion, and other DNA-related processes. In Bacillus subtilis, smc null mutations cause defects in DNA supercoiling, chromosome compaction, and chromosome partitioning. We investigated the effects of smc mutations on global gene expression in B. subtilis using DNA microarrays. We found that an smc null mutation caused partial induction of the SOS response, including induction of the defective prophage PBSX. Analysis of SOS and phage gene expression in single cells indicated that approximately 1% of smc mutants have fully induced SOS and PBSX gene expression while the other 99% of cells appear to have little or no expression. We found that induction of PBSX was not responsible for the chromosome partitioning or compaction defects of smc mutants. Similar inductions of the SOS response and PBSX were observed in cells depleted of topoisomerase I, an enzyme that relaxes negatively supercoiled DNA. PMID- 17416650 TI - Heterocyst development and diazotrophic metabolism in terminal respiratory oxidase mutants of the cyanobacterium Anabaena sp. strain PCC 7120. AB - Heterocyst development was analyzed in mutants of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 bearing inactivated cox2 and/or cox3 genes, encoding heterocyst-specific terminal respiratory oxidases. At the morphological level, the cox2 cox3 double mutant (strain CSAV141) was impaired in membrane reorganization involving the so-called honeycomb system that in the wild type strain is largely or exclusively devoted to respiration, accumulated glycogen granules at conspicuously higher levels than the wild type (in both vegetative cells and heterocysts), and showed a delay in carboxysome degradation upon combined nitrogen deprivation. Consistently, chemical analysis confirmed higher accumulation of glycogen in strain CSAV141 than in the wild type. No impairment was observed in the formation of the glycolipid or polysaccharide layers of the heterocyst envelope, consistent with the chemical detection of heterocyst-specific glycolipids, or in the expression of the heterocyst-specific genes nifHDK and fdxH. However, nitrogenase activity under oxic conditions was impaired in strain CSAV135 (cox3) and undetectable in strain CSAV141 (cox2 cox3). These results show that these dedicated oxidases are required for normal development and performance of the heterocysts and indicate a central role of Cox2 and, especially, of Cox3 in the respiratory activity of the heterocysts, decisively contributing to protection of the N(2) fixation machinery against oxygen. However, in contrast to the case for other diazotrophic bacteria, expression of nif genes in Anabaena seems not to be affected by oxygen. PMID- 17416651 TI - Role of RNA structure and susceptibility to RNase E in regulation of a cold shock mRNA, cspA mRNA. AB - Degradation of the cspA mRNA in vivo is very rapid at temperatures greater than 30 degrees C and is moderately dependent on RNase E. Investigations in vitro show that degradosomes prepared from normal or cold-shocked cultures cleave the cspA mRNA preferentially at a single site in vitro between two stem-loops approximately 24 residues 3' to the termination codon and approximately 31 residues from the 3' end. The site of cleavage is independent of the temperature and largely independent of the phosphorylation status of the 5' end of cspA mRNA. A 5' stem-loop, potential occlusion of the initiation and termination codons, temperature-dependent translational efficiency, and the position of the RNase E cleavage site can explain the differential stability of the cspA mRNA. PMID- 17416652 TI - SigmaE regulates and is regulated by a small RNA in Escherichia coli. AB - RybB is a small, Hfq-binding noncoding RNA originally identified in a screen of conserved intergenic regions in Escherichia coli. Fusions of the rybB promoter to lacZ were used to screen plasmid genomic libraries and genomic transposon mutants for regulators of rybB expression. A number of plasmids, including some carrying rybB, negatively regulated the fusion. An insertion in the rep helicase and one upstream of dnaK decreased expression of the fusion. Multicopy suppressors of these insertions led to identification of two plasmids that stimulated the fusion. One contained the gene for the response regulator OmpR; the second contained mipA, encoding a murein hydrolase. The involvement of MipA and OmpR in cell surface synthesis suggested that the rybB promoter might be dependent on sigma(E). The sequence upstream of the +1 of rybB contains a consensus sigma(E) promoter. The activity of rybB-lacZ was increased in cells lacking the RseA anti sigma factor and when sigma(E) was overproduced from a heterologous promoter. The activity of rybB-lacZ and the detection of RybB were totally abolished in an rpoE null strain. In vitro, sigma(E) efficiently transcribes from this promoter. Both a rybB mutation and an hfq mutation significantly increased expression of both rybB-lacZ and rpoE-lacZ fusions, consistent with negative regulation of the sigma(E) response by RybB and other small RNAs. Based on the plasmid screens, NsrR, a repressor sensitive to nitric oxide, was also found to negatively regulate sigma(E)-dependent promoters in an RseA-independent fashion. PMID- 17416653 TI - In vitro biosynthesis of ether-type glycolipids in the methanoarchaeon Methanothermobacter thermautotrophicus. AB - The biosynthesis of archaeal ether-type glycolipids was investigated in vitro using Methanothermobacter thermautotrophicus cell-free homogenates. The sole sugar moiety of glycolipids and phosphoglycolipids of the organism is the beta-D glucosyl-(1-->6)-D-glucosyl (gentiobiosyl) unit. The enzyme activities of archaeol:UDP-glucose beta-glucosyltransferase (monoglucosylarchaeol [MGA] synthase) and MGA:UDP-glucose beta-1,6-glucosyltransferase (diglucosylarchaeol [DGA] synthase) were found in the methanoarchaeon. The synthesis of DGA is probably a two-step glucosylation: (i) archaeol + UDP-glucose --> MGA + UDP, and (ii) MGA + UDP-glucose --> DGA + UDP. Both enzymes required the addition of K(+) ions and archaetidylinositol for their activities. DGA synthase was stimulated by 10 mM MgCl(2), in contrast to MGA synthase, which did not require Mg(2+). It was likely that the activities of MGA synthesis and DGA synthesis were carried out by different proteins because of the Mg(2+) requirement and their cellular localization. MGA synthase and DGA synthase can be distinguished in cell extracts greatly enriched for each activity by demonstrating the differing Mg(2+) requirements of each enzyme. MGA synthase preferred a lipid substrate with the sn 2,3 stereostructure of the glycerol backbone on which two saturated isoprenoid chains are bound at the sn-2 and sn-3 positions. A lipid substrate with unsaturated isoprenoid chains or sn-1,2-dialkylglycerol configuration exhibited low activity. Tetraether-type caldarchaetidylinositol was also actively glucosylated by the homogenates to form monoglucosyl caldarchaetidylinositol and a small amount of diglucosyl caldarchaetidylinositol. The addition of Mg(2+) increased the formation of diglucosyl caldarchaetidylinositol. This suggested that the same enzyme set synthesized the sole sugar moiety of diether-type glycolipids and tetraether-type phosphoglycolipids. PMID- 17416654 TI - Identification of the sigmaB regulon of Bacillus cereus and conservation of sigmaB-regulated genes in low-GC-content gram-positive bacteria. AB - The alternative sigma factor sigma(B) has an important role in the acquisition of stress resistance in many gram-positive bacteria, including the food-borne pathogen Bacillus cereus. Here, we describe the identification of the set of sigma(B)-regulated genes in B. cereus by DNA microarray analysis of the transcriptome upon a mild heat shock. Twenty-four genes could be identified as being sigma(B) dependent as witnessed by (i) significantly lower expression levels of these genes in mutants with a deletion of sigB and rsbY (which encode the alternative sigma factor sigma(B) and a crucial positive regulator of sigma(B) activity, respectively) than in the parental strain B. cereus ATCC 14579 and (ii) increased expression of these genes upon a heat shock. Newly identified sigma(B)-dependent genes in B. cereus include a histidine kinase and two genes that have predicted functions in spore germination. This study shows that the sigma(B) regulon of B. cereus is considerably smaller than that of other gram positive bacteria. This appears to be in line with phylogenetic analyses where sigma(B) of the B. cereus group was placed close to the ancestral form of sigma(B) in gram-positive bacteria. The data described in this study and previous studies in which the complete sigma(B) regulon of the gram-positive bacteria Bacillus subtilis, Listeria monocytogenes, and Staphylococcus aureus were determined enabled a comparison of the sets of sigma(B)-regulated genes in the different gram-positive bacteria. This showed that only three genes (rsbV, rsbW, and sigB) are conserved in their sigma(B) dependency in all four bacteria, suggesting that the sigma(B) regulon of the different gram-positive bacteria has evolved to perform niche-specific functions. PMID- 17416655 TI - Identification of essential residues in apolipoprotein N-acyl transferase, a member of the CN hydrolase family. AB - Apolipoprotein N-acyl transferase (Lnt) is an essential membrane-bound protein involved in lipid modification of all lipoproteins in gram-negative bacteria. Essential residues in Lnt of Escherichia coli were identified by using site directed mutagenesis and an in vivo complementation assay. Based on sequence conservation and known protein structures, we predict a model for Lnt, which is a member of the CN hydrolase family. Besides the potential catalytic triad E267 K335-C387, four residues that directly affect the modification of Braun's lipoprotein Lpp are absolutely required for Lnt function. Residues Y388 and E389 are part of the hydrophobic pocket that constitutes the active site. Residues W237 and E343 are located on two flexible arms that face away from the active site and are expected to open and close upon the binding and release of phospholipid and/or apolipoprotein. Substitutions causing temperature-dependent effects were located at different positions in the structural model. These mutants were not affected in protein stability. Lnt proteins from other proteobacteria, but not from actinomycetes, were functional in vivo, and the essential residues identified in Lnt of E. coli are conserved in these proteins. PMID- 17416656 TI - Molecular interaction between lipoteichoic acids and Lactobacillus delbrueckii phages depends on D-alanyl and alpha-glucose substitution of poly(glycerophosphate) backbones. AB - Lipoteichoic acids (LTAs) have been shown to act as bacterial counterparts to the receptor binding proteins of LL-H, LL-H host range mutant LL-H-a21, and JCL1032. Here we have used LTAs purified by hydrophobic interaction chromatography from different phage-resistant and -sensitive strains of Lactobacillus delbrueckii subsp. lactis. Nuclear magnetic resonance analyses revealed variation in the degree of alpha-glucosyl and D-alanyl substitution of the 1,3-linked poly(glycerophosphate) LTAs between the phage-sensitive and phage-resistant strains. Inactivation of phages was less effective if there was a high level of D alanine residues in the LTA backbones. Prior incubation of the LTAs with alpha glucose-specific lectin inhibited the LL-H phage inactivation. The overall level of decoration or the specific spatial combination of alpha-glucosyl-substituted, D-alanyl-substituted, and nonsubstituted glycerol residues may also affect phage adsorption. PMID- 17416657 TI - Distribution of protein A on the surface of Staphylococcus aureus. AB - Surface proteins of Staphylococcus aureus fulfill many important roles during the pathogenesis of human infections and are anchored to the cell wall envelope by sortases. Although the chemical linkage of proteins to cell wall cross bridges is known, the mechanisms whereby polypeptides are distributed on the staphylococcal surface have not been revealed. We show here that protein A, the ligand of immunoglobulin, is unevenly distributed over the staphylococcal surface. Upon removal with trypsin, newly synthesized polypeptide is deposited at two to four discrete foci. During subsequent growth, protein A appears to be slowly distributed from these sites. When viewed through multiple focal planes by laser scanning microscopy, protein A foci are arranged in a circle surrounding the bacterial cell. This pattern of distribution requires the LPXTG sorting signal of protein A as well as sortase A, the transpeptidase that anchors polypeptides to cell wall cross bridges. A model is presented whereby protein A deposition at discrete sites coupled with cell wall synthesis enables distribution of protein A on the staphylococcal surface. PMID- 17416658 TI - Purification and characterization of mycobacterial phospholipase A: an activity associated with mycobacterial cutinase. AB - We describe mycobacterial phospholipase A activity (MPLA) and, using reverse genetics, have associated this activity with putative mycobacterial cutinase. PLAs, which hydrolyze fatty acids on phospholipids, play a significant role in human inflammatory states and disease pathogenesis. In prokaryotes, the recognition of their role in virulence is more recent. Cutinases are serine esterases whose primary substrate is cutin, the waxy exterior layer of plants. Mycobacterium tuberculosis has maintained seven putative cutinases, though it should not encounter cutin; we demonstrate that known cutinases and MPLA cleave phospholipids in a PLA-type manner and also hydrolyze Tween. We analyzed cutinase motifs in mycobacteria and found the motif very prevalent. All mycobacteria tested had MPLA activity. These studies suggest an alternative use for putative cutinases by the M. tuberculosis group that is likely related to MPLA activity and lipid metabolism. PMID- 17416659 TI - The zinc-responsive regulator Zur controls a zinc uptake system and some ribosomal proteins in Streptomyces coelicolor A3(2). AB - In various bacteria, Zur, a zinc-specific regulator of the Fur family, regulates genes for zinc transport systems to maintain zinc homeostasis. It has also been suggested that Zur controls zinc mobilization by regulating some ribosomal proteins. The antibiotic-producing soil bacterium Streptomyces coelicolor contains four genes for Fur family regulators, and one (named zur) is located downstream of the znuACB operon encoding a putative zinc uptake transporter. We found that zinc specifically repressed the level of znuA transcripts and that this level was derepressed in a Delta zur mutant. Purified Zur existing as homodimers bound to the znuA promoter region in the presence of zinc, confirming the role of Zur as a zinc-responsive repressor. We analyzed transcripts for paralogous forms of ribosomal proteins L31 (RpmE1 and RpmE2) and L33 (RpmG2 and RpmG3) for their dependence on Zur and found that RpmE2 and RpmG2 with no zinc binding motif of conserved cysteines (C's) were negatively regulated by Zur. C negative RpmG3 and C-positive RpmE1 were not regulated by Zur. Instead, they were regulated by the sigma factor sigma(R) as predicted from their promoter sequences. The rpmE1 and rpmG3 genes were partially induced by EDTA in a manner dependent on sigma(R), suggesting that zinc depletion may stimulate the sigma(R) regulatory system. This finding reflects a link between thiol-oxidizing stress and zinc depletion. We determined the Zur-binding sites within znuA and rpmG2 promoter regions by footprinting analyses and identified a consensus inverted repeat sequence (TGaaAatgatTttCA, where uppercase letters represent the nucleotides common to all sites analyzed). This sequence closely matches that for mycobacterial Zur and allows the prediction of more genes in the Zur regulon. PMID- 17416660 TI - Characterization of a C-C bond hydrolase from Sphingomonas wittichii RW1 with novel specificities towards polychlorinated biphenyl metabolites. AB - Sphingomonas wittichii RW1 degrades chlorinated dibenzofurans and dibenzo-p dioxins via meta cleavage. We used inverse PCR to amplify dxnB2, a gene encoding one of three meta-cleavage product (MCP) hydrolases identified in the organism that are homologues of BphD involved in biphenyl catabolism. Purified DxnB2 catalyzed the hydrolysis of 8-OH 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate (HOPDA) approximately six times faster than for HOPDA at saturating substrate concentrations. Moreover, the specificity of DxnB2 for HOPDA (k(cat)/K(m) = 1.2 x 10(7) M(-1) s(-1)) was about half that of the BphDs of Burkholderia xenovorans LB400 and Rhodococcus globerulus P6, two potent polychlorinated biphenyl (PCB) degrading strains. Interestingly, DxnB2 transformed 3-Cl and 4-OH HOPDAs, compounds that inhibit the BphDs and limit PCB degradation. DxnB2 had a higher specificity for 9-Cl HOPDA than for HOPDA but a lower specificity for 8-Cl HOPDA (k(cat)/K(m) = 1.7 x 10(6) M(-1) s(-1)), the chlorinated analog of 8-OH HOPDA produced during dibenzofuran catabolism. Phylogenetic analyses based on structure guided sequence alignment revealed that DxnB2 belongs to a previously unrecognized class of MCP hydrolases, evolutionarily divergent from the BphDs although the physiological substrates of both enzyme types are HOPDAs. However, both classes of enzymes have mainly small hydrophobic residues lining the subsite that binds the C-6 phenyl of HOPDA, in contrast to the bulky hydrophobic residues (Phe106, Phe135, Trp150, and Phe197) found in the class II enzymes that prefer substrates possessing a C-6 alkyl. Thr196 and/or Asn203 appears to be an important determinant of specificity for DxnB2, potentially forming hydrogen bonds with the 8-OH substituent. This study demonstrates that the substrate specificities of evolutionarily divergent hydrolases may be useful for degrading mixtures of pollutants, such as PCBs. PMID- 17416661 TI - Citrate sensing by the C4-dicarboxylate/citrate sensor kinase DcuS of Escherichia coli: binding site and conversion of DcuS to a C4-dicarboxylate- or citrate specific sensor. AB - The histidine protein kinase DcuS of Escherichia coli senses C(4)-dicarboxylates and citrate by a periplasmic domain. The closely related sensor kinase CitA binds citrate, but no C(4)-dicarboxylates, by a homologous periplasmic domain. CitA is known to bind the three carboxylate and the hydroxyl groups of citrate by sites C1, C2, C3, and H. DcuS requires the same sites for C(4)-dicarboxylate sensing, but only C2 and C3 are highly conserved. It is shown here that sensing of citrate by DcuS required the same sites. Binding of citrate to DcuS, therefore, was similar to binding of C(4)-dicarboxylates but different from that of citrate binding in CitA. DcuS could be converted to a C(4)-dicarboxylate-specific sensor (DcuS(DC)) by mutating residues of sites C1 and C3 or of some DcuS-subtype specific residues. Mutations around site C1 aimed at increasing the size and accessibility of the site converted DcuS to a citrate-specific sensor (DcuS(Cit)). DcuS(DC) and DcuS(Cit) had complementary effector specificities and responded either to C(4)-dicarboxylates or to citrate and mesaconate. The results imply that DcuS binds citrate (similar to the C(4)-dicarboxylates) via the C(4) dicarboxylate part of the molecule. Sites C2 and C3 are essential for binding of two carboxylic groups of citrate or of C(4)-dicarboxylates; sites C1 and H are required for other essential purposes. PMID- 17416662 TI - Flagellar motility and structure in the hyperthermoacidophilic archaeon Sulfolobus solfataricus. AB - Flagellation in archaea is widespread and is involved in swimming motility. Here, we demonstrate that the structural flagellin gene from the crenarchaeaon Sulfolobus solfataricus is highly expressed in stationary-phase-grown cells and under unfavorable nutritional conditions. A mutant in a flagellar auxiliary gene, flaJ, was found to be nonmotile. Electron microscopic imaging of the flagellum indicates that the filaments are composed of right-handed helices. PMID- 17416663 TI - Colicin E2 is still in contact with its receptor and import machinery when its nuclease domain enters the cytoplasm. AB - Colicins reach their targets in susceptible Escherichia coli strains through two envelope protein systems: the Tol system is used by group A colicins and the TonB system by group B colicins. Colicin E2 (ColE2) is a cytotoxic protein that recognizes the outer membrane receptor BtuB. After gaining access to the cytoplasmic membrane of sensitive Escherichia coli cells, ColE2 enters the cytoplasm to cleave DNA. After binding to BtuB, ColE2 interacts with the Tol system to reach its target. However, it is not known if the entire colicin or only the nuclease domain of ColE2 enters the cell. Here I show that preincubation of ColE2 with Escherichia coli cells prevents binding and translocation of pore forming colicins of group A but not of group B. This inhibition persisted even when cells were incubated with ColE2 for 30 min before the addition of pore forming colicins, indicating that ColE2 releases neither its receptor nor its translocation machinery when its nuclease domain enters the cells. These competition experiments enabled me to estimate the time required for ColE2 binding to its receptor and translocation. PMID- 17416664 TI - The phosphotransferase system formed by PtsP, PtsO, and PtsN proteins controls production of polyhydroxyalkanoates in Pseudomonas putida. AB - The genome of Pseudomonas putida KT2440 encodes five proteins of the phosphoenolpyruvate-carbohydrate phosphotransferase system. Two of these (FruA and FruB) form a dedicated system for fructose intake, while enzyme I(Ntr) (EI(Ntr); encoded by ptsP), NPr (ptsO), and EII(Ntr) (ptsN) act in concert to control the intracellular accumulation of polyhydroxyalkanoates, a typical product of carbon overflow. PMID- 17416665 TI - Characterization of a novel bifunctional dihydropteroate synthase/dihydropteroate reductase enzyme from Helicobacter pylori. AB - Tetrahydrofolate is a ubiquitous C(1) carrier in many biosynthetic pathways in bacteria, importantly, in the biosynthesis of formylmethionyl tRNA(fMet), which is essential for the initiation of translation. The final step in the biosynthesis of tetrahydrofolate is carried out by the enzyme dihydrofolate reductase (DHFR). A search of the complete genome sequence of Helicobacter pylori failed to reveal any sequence that encodes DHFR. Previous studies demonstrated that the H. pylori dihydropteroate synthase gene folP can complement an Escherichia coli strain in which folA and folM, encoding two distinct DHFRs, are deleted. It was also shown that H. pylori FolP possesses an additional N-terminal domain that binds flavin mononucleotide (FMN). Homologous domains are found in FolP proteins of other microorganisms that do not possess DHFR. In this study, we demonstrated that H. pylori FolP is also a dihydropteroate reductase that derives its reducing power from soluble flavins, reduced FMN and reduced flavin adenine dinucleotide. We also determined the stoichiometry of the enzyme-bound flavin and showed that half of the bound flavin is exchangeable with the soluble flavins. Finally, site-directed mutagenesis of the most conserved amino acid residues in the N-terminal domain indicated the importance of these residues for the activity of the enzyme as a dihydropteroate reductase. PMID- 17416666 TI - Characterization of a small mobilizable transposon, MTnSag1, in Streptococcus agalactiae. AB - The transposon MTnSag1 from Streptococcus agalactiae carried an IS1-like transposase gene and the lnu(C) gene, which encoded a lincosamide nucleotidyltransferase. MTnSag1 could be mobilized by the conjugative transposon Tn916. An intermediate circular form of MTnSag1 and a putative origin of transfer at the 3' end of the lnu(C) gene were characterized. PMID- 17416667 TI - Genome of Methylobacillus flagellatus, molecular basis for obligate methylotrophy, and polyphyletic origin of methylotrophy. AB - Along with methane, methanol and methylated amines represent important biogenic atmospheric constituents; thus, not only methanotrophs but also nonmethanotrophic methylotrophs play a significant role in global carbon cycling. The complete genome of a model obligate methanol and methylamine utilizer, Methylobacillus flagellatus (strain KT) was sequenced. The genome is represented by a single circular chromosome of approximately 3 Mbp, potentially encoding a total of 2,766 proteins. Based on genome analysis as well as the results from previous genetic and mutational analyses, methylotrophy is enabled by methanol and methylamine dehydrogenases and their specific electron transport chain components, the tetrahydromethanopterin-linked formaldehyde oxidation pathway and the assimilatory and dissimilatory ribulose monophosphate cycles, and by a formate dehydrogenase. Some of the methylotrophy genes are present in more than one (identical or nonidentical) copy. The obligate dependence on single-carbon compounds appears to be due to the incomplete tricarboxylic acid cycle, as no genes potentially encoding alpha-ketoglutarate, malate, or succinate dehydrogenases are identifiable. The genome of M. flagellatus was compared in terms of methylotrophy functions to the previously sequenced genomes of three methylotrophs, Methylobacterium extorquens (an alphaproteobacterium, 7 Mbp), Methylibium petroleiphilum (a betaproteobacterium, 4 Mbp), and Methylococcus capsulatus (a gammaproteobacterium, 3.3 Mbp). Strikingly, metabolically and/or phylogenetically, the methylotrophy functions in M. flagellatus were more similar to those in M. capsulatus and M. extorquens than to the ones in the more closely related M. petroleiphilum species, providing the first genomic evidence for the polyphyletic origin of methylotrophy in Betaproteobacteria. PMID- 17416668 TI - Characterization of an arginine:pyruvate transaminase in arginine catabolism of Pseudomonas aeruginosa PAO1. AB - The arginine transaminase (ATA) pathway represents one of the multiple pathways for L-arginine catabolism in Pseudomonas aeruginosa. The AruH protein was proposed to catalyze the first step in the ATA pathway, converting the substrates L-arginine and pyruvate into 2-ketoarginine and L-alanine. Here we report the initial biochemical characterization of this enzyme. The aruH gene was overexpressed in Escherichia coli, and its product was purified to homogeneity. High-performance liquid chromatography and mass spectrometry (MS) analyses were employed to detect the presence of the transamination products 2-ketoarginine and L-alanine, thus demonstrating the proposed biochemical reaction catalyzed by AruH. The enzymatic properties and kinetic parameters of dimeric recombinant AruH were determined by a coupled reaction with NAD(+) and L-alanine dehydrogenase. The optimal activity of AruH was found at pH 9.0, and it has a novel substrate specificity with an order of preference of Arg > Lys > Met > Leu > Orn > Gln. With L-arginine and pyruvate as the substrates, Lineweaver-Burk plots of the data revealed a series of parallel lines characteristic of a ping-pong kinetic mechanism with calculated V(max) and k(cat) values of 54.6 +/- 2.5 micrromol/min/mg and 38.6 +/- 1.8 s(-1). The apparent K(m) and catalytic efficiency (k(cat)/K(m)) were 1.6 +/- 0.1 mM and 24.1 mM(-1) s(-1) for pyruvate and 13.9 +/- 0.8 mM and 2.8 mM(-1) s(-1) for l-arginine. When L-lysine was used as the substrate, MS analysis suggested Delta(1)-piperideine-2-carboxylate as its transamination product. These results implied that AruH may have a broader physiological function in amino acid catabolism. PMID- 17416669 TI - Interspecies DNA microarray analysis identifies WblA as a pleiotropic down regulator of antibiotic biosynthesis in Streptomyces. AB - Using Streptomyces coelicolor microarrays to discover regulators of gene expression in other Streptomyces species, we identified wblA, a whiB-like gene encoding a putative transcription factor, as a down-regulator of doxorubicin biosynthesis in Streptomyces peucetius. Further analysis revealed that wblA functions pleiotropically to control antibiotic production and morphological differentiation in streptomycetes. Our results reveal a novel biological role for wblA and show the utility of interspecies microarray analysis for the investigation of streptomycete gene expression. PMID- 17416670 TI - Functional genomics enables identification of genes of the arginine transaminase pathway in Pseudomonas aeruginosa. AB - Arginine utilization in Pseudomonas aeruginosa with multiple catabolic pathways represents one of the best examples of the metabolic versatility of this organism. To identify genes involved in arginine catabolism, we have employed DNA microarrays to analyze the transcriptional profiles of this organism in response to L-arginine. While most of the genes involved in arginine uptake, regulation, and metabolism have been identified as members of the ArgR (arginine-responsive regulatory protein) regulon in our previous study, they did not include any genes of the arginine dehydrogenase (ADH) pathway. In this study, 18 putative transcriptional units of 38 genes, including the two known genes of the ADH pathway, kauB and gbuA, were found to be inducible by exogenous L-arginine in the absence of ArgR. To identify the missing genes that encode enzymes for the initial steps of the ADH pathway, the potential physiological functions of those candidate genes in arginine utilization were studied by growth phenotype analysis of knockout mutants. Expression of these genes was induced by L-arginine in an aruF mutant strain devoid of a functional arginine succinyltransferase pathway, the major route of arginine utilization. Disruption of dadA, a putative catabolic alanine dehydrogenase-encoding gene, in the aruF mutant produced no growth on L arginine, suggesting the involvement of L-alanine in arginine catabolism. This hypothesis was further supported by the detection of an L-arginine-inducible arginine:pyruvate transaminase activity in the aruF mutant. Knockout of aruH and aruI, which encode an arginine:pyruvate transaminase and a 2-ketoarginine decarboxylase in an operon, also abolished the ability of the aruF mutant to grow on L-arginine. The results of high-performance liquid chromatography analysis demonstrated consumption of 2-ketoarginine and suggested that generation of 4 guanidinobutyraldehyde occurred in the aruF mutant but not in the aruF aruI mutant. These results led us to propose the arginine transaminase pathway that removes the alpha-amino group of L-arginine via transamination instead of oxidative deamination by dehydrogenase or oxidase as originally proposed. In the same genetic locus, we also identified a two-component system, AruRS, for the regulation of arginine-responsive induction of the arginine transaminase pathway. This work depicted a wider network of arginine metabolism than we previously recognized. PMID- 17416671 TI - Eukaryotic-type serine/threonine protein kinase StkP is a global regulator of gene expression in Streptococcus pneumoniae. AB - Signal transduction pathways in both prokaryotes and eukaryotes utilize protein phosphorylation as a key regulatory mechanism. Recent studies have proven that eukaryotic-type serine/threonine protein kinases (Hank's type) are widespread in many bacteria, although little is known regarding the cellular processes they control. In this study, we have attempted to establish the role of a single eukaryotic-type protein kinase, StkP of Streptococcus pneumoniae, in bacterial survival. Our results indicate that the expression of StkP is important for the resistance of S. pneumoniae to various stress conditions. To investigate the impact of StkP on this phenotype, we compared the whole-genome expression profiles of the wild-type and DeltastkP mutant strains by microarray technology. This analysis revealed that StkP positively controls the transcription of a set of genes encoding functions involved in cell wall metabolism, pyrimidine biosynthesis, DNA repair, iron uptake, and oxidative stress response. Despite the reduced transformability of the stkP mutant, we found that the competence regulon was derepressed in the stkP mutant under conditions that normally repress natural competence development. Furthermore, the competence regulon was expressed independently of exogenous competence-stimulating peptide. In summary, our studies show that a eukaryotic-type serine/threonine protein kinase functions as a global regulator of gene expression in S. pneumoniae. PMID- 17416672 TI - An early modern human from Tianyuan Cave, Zhoukoudian, China. AB - Thirty-four elements of an early modern human (EMH) were found in Tianyuan Cave, Zhoukoudian, China in 2003. Dated to 42,000-39,000 calendrical years before present by using direct accelerator mass spectrometry radiocarbon, the Tianyuan 1 skeleton is among the oldest directly dated EMHs in eastern Eurasia. Morphological comparison shows Tianyuan 1 to have a series of derived modern human characteristics, including a projecting tuber symphyseos, a high anterior symphyseal angle, a broad scapular glenoid fossa, a reduced hamulus, a gluteal buttress, and a pilaster on the femora. Other features of Tianyuan 1 that are more common among EMHs are its modest humeral pectoralis major tuberosities, anteriorly rotated radial tuberosity, reduced radial curvature, and modest talar trochlea. It also lacks several mandibular features common among western Eurasian late archaic humans, including mandibular foramen bridging, mandibular notch asymmetry, and a large superior medial pterygoid tubercle. However, Tianyuan 1 exhibits several late archaic human features, such as its anterior to posterior dental proportions, a large hamulus length, and a broad and rounded distal phalangeal tuberosity. This morphological pattern implies that a simple spread of modern humans from Africa is unlikely. PMID- 17416673 TI - Network of coregulated spliceosome components revealed by zebrafish mutant in recycling factor p110. AB - The spliceosome cycle consists of assembly, catalysis, and recycling phases. Recycling of postspliceosomal U4 and U6 small nuclear ribonucleoproteins (snRNPs) requires p110/SART3, a general splicing factor. In this article, we report that the zebrafish earl grey (egy) mutation maps in the p110 gene and results in a phenotype characterized by thymus hypoplasia, other organ-specific defects, and death by 7 to 8 days postfertilization. U4/U6 snRNPs were disrupted in egy mutant embryos, demonstrating the importance of p110 for U4/U6 snRNP recycling in vivo. Surprisingly, expression profiling of the egy mutant revealed an extensive network of coordinately up-regulated components of the spliceosome cycle, providing a mechanism compensating for the recycling defect. Together, our data demonstrate that a mutation in a general splicing factor can lead to distinct defects in organ development and cause disease. PMID- 17416674 TI - Group selection and kin selection: two concepts but one process. AB - In a recent paper, Traulsen and Nowak use a multilevel selection model to show that cooperation can be favored by group selection in finite populations [Traulsen A, Nowak M (2006) Proc Natl Acad Sci USA 103:10952-10955]. The authors challenge the view that kin selection may be an appropriate interpretation of their results and state that group selection is a distinctive process "that permeates evolutionary processes from the emergence of the first cells to eusociality and the economics of nations." In this paper, we start by addressing Traulsen and Nowak's challenge and demonstrate that all their results can be obtained by an application of kin selection theory. We then extend Traulsen and Nowak's model to life history conditions that have been previously studied. This allows us to highlight the differences and similarities between Traulsen and Nowak's model and typical kin selection models and also to broaden the scope of their results. Our retrospective analyses of Traulsen and Nowak's model illustrate that it is possible to convert group selection models to kin selection models without disturbing the mathematics describing the net effect of selection on cooperation. PMID- 17416675 TI - The alanine-rich XAO peptide adopts a heterogeneous population, including turn like and polyproline II conformations. AB - The solution structure of the hepta-alanine polypeptide Ac-X(2)A(7)O(2)-NH(2) (XAO) has been a matter of controversy in the current literature. On one side of the argument is a claim that the peptide adopts a mostly polyproline II (PPII) structure, with a <20% population of beta conformations at room temperature [Shi Z, Olson CA, Rose GA, Baldwin RL, Kallenbach NR (2002) Proc Natl Acad Sci USA 99:9190-9195], whereas the other side of the argument insists that the peptide exists as an ensemble of conformations, including multiple beta-turn structures [Makowska J, Rodziewicz-Motowidlo S, Baginska K, Vila JA, Liwo A, Chmurzynski L, Scheraga HA (2006) Proc Natl Acad Sci USA 103:1744-1749]. We have used an excitonic coupling model to simulate the amide I band of the FTIR, vibrational circular dichroism, and isotropic and anisotropic Raman spectra of XAO, where, for each residue, the backbone dihedral angle varphi was constrained by using the reported (3)J(CalphaHNH) values and a modified Karplus relation. The best reproduction of the experimental data could only be achieved by assuming an ensemble of conformations, which contains various beta-turn conformations ( approximately 26%), in addition to beta-strand ( approximately 23%) and PPII ( approximately 50%) conformations. PPII is the dominant conformation in segments not involved in turn formations. Most of the residues were found to sample the bridge region connecting the PPII and right-handed helix troughs in the Ramachandran plot, which is part of the very heterogeneous ensemble of conformations generally termed type IV beta-turn. PMID- 17416676 TI - Symmetry matters for the electronic structure of core complexes from Rhodopseudomonas palustris and Rhodobacter sphaeroides PufX-. AB - Low-temperature (1.4 K), single-molecule fluorescence-excitation spectra have been recorded for individual reaction center-light-harvesting 1 complexes from Rhodopseudomonas palustris and the PufX(-) strain of Rhodobacter sphaeroides. More than 80% of the complexes from Rb. sphaeroides show only broad absorption bands, whereas nearly all of the complexes from Rps. palustris also have a narrow line at the low-energy end of their spectrum. We describe how the presence of this narrow feature indicates the presence of a gap in the electronic structure of the light-harvesting 1 complex from Rps. palustris, which provides strong support for the physical gap that was previously modeled in its x-ray crystal structure. PMID- 17416677 TI - The effect of public health measures on the 1918 influenza pandemic in U.S. cities. AB - During the 1918 influenza pandemic, the U.S., unlike Europe, put considerable effort into public health interventions. There was also more geographic variation in the autumn wave of the pandemic in the U.S. compared with Europe, with some cities seeing only a single large peak in mortality and others seeing double peaked epidemics. Here we examine whether differences in the public health measures adopted by different cities can explain the variation in epidemic patterns and overall mortality observed. We show that city-specific per-capita excess mortality in 1918 was significantly correlated with 1917 per-capita mortality, indicating some intrinsic variation in overall mortality, perhaps related to sociodemographic factors. In the subset of 23 cities for which we had partial data on the timing of interventions, an even stronger correlation was found between excess mortality and how early in the epidemic interventions were introduced. We then fitted an epidemic model to weekly mortality in 16 cities with nearly complete intervention-timing data and estimated the impact of interventions. The model reproduced the observed epidemic patterns well. In line with theoretical arguments, we found the time-limited interventions used reduced total mortality only moderately (perhaps 10-30%), and that the impact was often very limited because of interventions being introduced too late and lifted too early. San Francisco, St. Louis, Milwaukee, and Kansas City had the most effective interventions, reducing transmission rates by up to 30-50%. Our analysis also suggests that individuals reactively reduced their contact rates in response to high levels of mortality during the pandemic. PMID- 17416678 TI - The role of alkane coordination in C-H bond cleavage at a Pt(II) center. AB - The rates of C H bond activation for various alkanes by [(N-N)Pt(Me)(TFEd(3))](+) (N N = Ar N C(Me) C(Me) N Ar; Ar = 3,5-di-tert-butylphenyl; TFE-d(3) = CF(3)CD(2)OD) were studied. Both linear and cyclic alkanes give the corresponding alkene-hydride cation [(N-N)Pt(H)(alkene)](+) via (i) rate determining alkane coordination to form a C H sigma complex, (ii) oxidative cleavage of the coordinated C H bond to give a platinum(IV) alkyl-methyl-hydride intermediate, (iii) reductive coupling to generate a methane sigma complex, (iv) dissociation of methane, and (v) beta-H elimination to form the observed product. Second-order rate constants for cycloalkane activation (C(n)H(2n)), are proportional to the size of the ring (k approximately n). For cyclohexane, the deuterium kinetic isotope effect (k(H)/k(D)) of 1.28 (5) is consistent with the proposed rate determining alkane coordination to form a C H sigma complex. Statistical scrambling of the five hydrogens of the Pt-methyl and the coordinated methylene unit, via rapid, reversible steps ii and iii, and interchange of geminal C H bonds of the methane and cyclohexane C H sigma adducts, is observed before loss of methane. PMID- 17416679 TI - Public health interventions and epidemic intensity during the 1918 influenza pandemic. AB - Nonpharmaceutical interventions (NPIs) intended to reduce infectious contacts between persons form an integral part of plans to mitigate the impact of the next influenza pandemic. Although the potential benefits of NPIs are supported by mathematical models, the historical evidence for the impact of such interventions in past pandemics has not been systematically examined. We obtained data on the timing of 19 classes of NPI in 17 U.S. cities during the 1918 pandemic and tested the hypothesis that early implementation of multiple interventions was associated with reduced disease transmission. Consistent with this hypothesis, cities in which multiple interventions were implemented at an early phase of the epidemic had peak death rates approximately 50% lower than those that did not and had less steep epidemic curves. Cities in which multiple interventions were implemented at an early phase of the epidemic also showed a trend toward lower cumulative excess mortality, but the difference was smaller (approximately 20%) and less statistically significant than that for peak death rates. This finding was not unexpected, given that few cities maintained NPIs longer than 6 weeks in 1918. Early implementation of certain interventions, including closure of schools, churches, and theaters, was associated with lower peak death rates, but no single intervention showed an association with improved aggregate outcomes for the 1918 phase of the pandemic. These findings support the hypothesis that rapid implementation of multiple NPIs can significantly reduce influenza transmission, but that viral spread will be renewed upon relaxation of such measures. PMID- 17416680 TI - Insulin receptors in beta-cells are critical for islet compensatory growth response to insulin resistance. AB - Insulin and insulin-like growth factor 1 (IGF1) are ubiquitous growth factors that regulate proliferation in most mammalian tissues including pancreatic islets. To explore the specificity of insulin receptors in compensatory beta-cell growth, we examined two models of insulin resistance. In the first model, we used liver-specific insulin receptor knockout (LIRKO) mice, which exhibit hyperinsulinemia without developing diabetes due to a compensatory increase in beta-cell mass. LIRKO mice, also lacking functional insulin receptors in beta cells (beta IRKO/LIRKO), exhibited severe glucose intolerance but failed to develop compensatory islet hyperplasia, together leading to early death. In the second model, we examined the relative significance of insulin versus IGF1 receptors in islet growth by feeding high-fat diets to beta IRKO and beta-cell specific IGF1 receptor knockout (beta IGFRKO) mice. Although both groups on the high-fat diet developed insulin resistance, beta IRKO, but not beta IGFRKO, mice exhibited poor islet growth consistent with insulin-stimulated phosphorylation, nuclear exclusion of FoxO1, and reduced expression of Pdx-1. Together these data provide direct genetic evidence that insulin/FoxO1/Pdx-1 signaling is one pathway that is crucial for islet compensatory growth response to insulin resistance. PMID- 17416681 TI - Tale of two spikes in bacteriophage PRD1. AB - Structural comparisons between bacteriophage PRD1 and adenovirus have revealed an evolutionary relationship that has contributed significantly to current ideas on virus phylogeny. However, the structural organization of the receptor-binding spike complex and how the different symmetry mismatches are mediated between the spike-complex proteins are not clear. We determined the architecture of the PRD1 spike complex by using electron microscopy and three-dimensional image reconstruction of a series of PRD1 mutants. We constructed an atomic model for the full-length P5 spike protein by using comparative modeling. P5 was shown to be bound directly to the penton base protein P31. P5 and the receptor-binding protein P2 form two separate spikes, interacting with each other near the capsid shell. P5, with a tumor necrosis factor-like head domain, may have been responsible for host recognition before capture of the current receptor-binding protein P2. PMID- 17416682 TI - The species, sex, and stage specificity of a Caenorhabditis sex pheromone. AB - Four species in the ELEGANS group of subgenus the Caenorhabditis are distinguished by two very different mating systems: androdioecy in C. elegans and Caenorhabditis briggsae with males and self-fertilizing hermaphrodites and dioecy in Caenorhabditis remanei and Caenorhabditis sp. strain CB5161 with males and females. Using chemotaxis assays, we demonstrate that females secrete a potent sex pheromone that attracts males from a distance, whereas hermaphrodites do not. The female sex pheromone is not species-specific, with males of all four species attracted to both the C. remanei and Caenorhabditis sp. female sex pheromones. The pheromone is, however, sex-specific, with only females secreting the pheromone and attracting only males. Furthermore, the sex pheromone is stage specific, with female secretion and male detection of the pheromone beginning near adulthood. Females lose their attractiveness immediately after mating but regain it several hours after mating ceases. Finally, the female somatic gonad is required for sex-pheromone production, and the male-specific cephalic neurons (CEM) are required for male response. PMID- 17416683 TI - Recombinant Saccharomyces cerevisiae strain expressing a model cytochrome P450 in the rat digestive environment: viability and bioconversion activity. AB - An innovative "biodrug" concept, based on the oral administration of living recombinant microorganisms, has recently emerged for the prevention or treatment of various diseases. An engineered Saccharomyces cerevisiae strain expressing plant P450 73A1 (cinnamate-4-hydroxylase [CA4H] activity) was used, and its survival and ability to convert trans-cinnamic acid (CIN) into p-coumaric acid (COU) were investigated in vivo. In rats, the recombinant yeast was resistant to gastric and small intestinal secretions but was more sensitive to the conditions found in the large intestine. After oral administration of yeast and CIN, the CA4H activity was shown in vivo, with COU being found throughout the rat's digestive tract and in its urine. The bioconversion reaction occurred very fast, with most of the COU being produced within the first 5 min. The gastrointestinal sac technique demonstrated that the recombinant yeast was able to convert CIN into COU (conversion rate ranging from 2 to 5%) in all the organs of the rat's digestive tract: stomach, duodenum, jejunum, ileum, cecum, and colon. These results promise new opportunities for the development of drug delivery systems based on engineered yeasts catalyzing a bioconversion reaction directly in the digestive tract. PMID- 17416684 TI - Clonothrix fusca Roze 1896, a filamentous, sheathed, methanotrophic gamma proteobacterium. AB - Crenothrix polyspora Cohn 1870 and Clonothrix fusca Roze 1896 are two filamentous, sheathed microorganisms exhibiting complex morphological differentiation, whose phylogeny and physiology have been obscure for a long time due to the inability to cultivate them. Very recently, DNA sequencing data from uncultured C. polyspora-enriched material have suggested that Crenothrix is a methane-oxidizing gamma-proteobacterium (39). In contrast, the possible ecological function of C. fusca, originally considered a developmental stage of C. polyspora, is unknown. In this study, temporal succession of two filamentous, sheathed microorganisms resembling Cohn's Crenothrix and Roze's Clonothrix was observed by analyzing the microbial community of an artesian well by optical microscopy. Combined culture-based and culture-independent approaches enabled us to assign C. fusca to a novel subgroup of methane-oxidizing gamma-proteobacteria distinct from that of C. polyspora. This assignment was supported by (i) methane uptake and assimilation experiments, (ii) ultrastructural data showing the presence in C. fusca cytoplasm of an elaborate membrane system resembling that of methanotrophic gamma-proteobacteria, and (iii) sequencing data demonstrating the presence in its genome of a methanol dehydrogenase alpha subunit-encoding gene (mxaF) and a conventional particulate methane mono-oxygenase alpha subunit encoding gene (pmoA) that is different from the unusual pmoA (u-pmoA) of C. polyspora. PMID- 17416685 TI - Evaluation of two surface sampling methods for detection of Erwinia herbicola on a variety of materials by culture and quantitative PCR. AB - This research was designed to evaluate surface sampling protocols for use with culture and quantitative PCR (QPCR) amplification assay for detection of the gram negative bacterial biothreat simulant Erwinia herbicola on a variety of surface materials. Surfaces selected for evaluation were wood laminate, glass and computer monitor screens, metal file cabinets, plastic arena seats, nylon seat cushions, finished concrete flooring, and vinyl tile flooring. Laboratory and test chamber studies were performed to evaluate two sampling methods, a sponge and a macrofoam swab, for detection of E. herbicola on surface materials. In laboratory trials, seven materials were inoculated with a known concentration of E. herbicola cells and samples were collected from the surfaces of the materials to determine sampling efficiencies. Culture analysis was ineffective for assessing E. herbicola collection efficiency because very few culturable cells were obtained from surface samples. QPCR demonstrated that E. herbicola DNA was present in high concentrations on all of the surface samples, and sampling efficiencies ranged from 0.7 to 52.2%, depending on the sampling method and the surface material. The swab was generally more efficient than the sponge for collection of E. herbicola from surfaces. Test chamber trials were also performed in which E. herbicola was aerosolized into the chamber and allowed to settle onto test materials. Surface sampling results supported those obtained in laboratory trials. The results of this study demonstrate the capabilities of QPCR to enhance the detection and enumeration of biocontaminants on surface materials and provide information on the comparability of sampling methods. PMID- 17416686 TI - Ammonia-oxidizing beta-proteobacteria from the oxygen minimum zone off northern Chile. AB - The composition of ammonia-oxidizing bacteria from the beta-Proteobacteria subclass (betaAOB) was studied in the surface and upper-oxycline oxic waters (2- to 50-m depth, approximately 200 to 44 microM O(2)) and within the oxygen minimum zone (OMZ) suboxic waters (50- to 400-m depth, < or =10 microM O(2)) of the eastern South Pacific off northern Chile. This study was carried out through cloning and sequencing of genes coding for 16S rRNA and the ammonia monooxygenase enzyme active subunit (amoA). Sequences affiliated with Nitrosospira-like cluster 1 dominated the 16S rRNA gene clone libraries constructed from both oxic and suboxic waters. Cluster 1 consists exclusively of yet-uncultivated betaAOB from marine environments. However, a single clone, out of 224 obtained from the OMZ, was found to belong to Nitrosospira lineage cluster 0. To our knowledge, cluster 0 sequences have been derived from betaAOB isolated only from sand, soil, and freshwater environments. Sequences in clone libraries of the amoA gene from the surface and upper oxycline could be grouped in a marine subcluster, also containing no cultured representatives. In contrast, all 74 amoA sequences originating from the OMZ were either closely affiliated with cultured Nitrosospira spp. from clusters 0 and 2 or with other yet-uncultured betaAOB from soil and an aerated-anoxic Orbal process waste treatment plant. Our results reveal the presence of Nitrosospira-like betaAOB in both oxic and suboxic waters associated with the OMZ but with a clear community shift at the functional level (amoA) along the strong oxygen gradient. PMID- 17416687 TI - Regulation of caffeate respiration in the acetogenic bacterium Acetobacterium woodii. AB - The anaerobic acetogenic bacterium Acetobacterium woodii can conserve energy by oxidation of various substrates coupled to either carbonate or caffeate respiration. We used a cell suspension system to study the regulation and kinetics of induction of caffeate respiration. After addition of caffeate to suspensions of fructose-grown cells, there was a lag phase of about 90 min before caffeate reduction commenced. However, in the presence of tetracycline caffeate was not reduced, indicating that de novo protein synthesis is required for the ability to respire caffeate. Induction also took place in the presence of CO(2), and once a culture was induced, caffeate and CO(2) were used simultaneously as electron acceptors. Induction of caffeate reduction was also observed with H(2) plus CO(2) as the substrate, but the lag phase was much longer. Again, caffeate and CO(2) were used simultaneously as electron acceptors. In contrast, during oxidation of methyl groups derived from methanol or betaine, acetogenesis was the preferred energy-conserving pathway, and caffeate reduction started only after acetogenesis was completed. The differential flow of reductants was also observed with suspensions of resting cells in which caffeate reduction was induced prior to harvest of the cells. These cell suspensions utilized caffeate and CO(2) simultaneously with fructose or hydrogen as electron donors, but CO(2) was preferred over caffeate during methyl group oxidation. Caffeate-induced resting cells could reduce caffeate and also p-coumarate or ferulate with hydrogen as the electron donor. p-Coumarate or ferulate also served as an inducer for caffeate reduction. Interestingly, caffeate-induced cells reduced ferulate in the absence of an external reductant, indicating that caffeate also induces the enzymes required for oxidation of the methyl group of ferulate. PMID- 17416688 TI - Epilithic cyanobacterial communities of a marine tropical beach rock (Heron Island, Great Barrier Reef): diversity and diazotrophy. AB - The diversity and nitrogenase activity of epilithic marine microbes in a Holocene beach rock (Heron Island, Great Barrier Reef, Australia) with a proposed biological calcification "microbialite" origin were examined. Partial 16S rRNA gene sequences from the dominant mat (a coherent and layered pink-pigmented community spread over the beach rock) and biofilms (nonstratified, differently pigmented microbial communities of small shallow depressions) were retrieved using denaturing gradient gel electrophoresis (DGGE), and a clone library was retrieved from the dominant mat. The 16S rRNA gene sequences and morphological analyses revealed heterogeneity in the cyanobacterial distribution patterns. The nonheterocystous filamentous genus Blennothrix sp., phylogenetically related to Lyngbya, dominated the mat together with unidentified nonheterocystous filaments of members of the Pseudanabaenaceae and the unicellular genus Chroococcidiopsis. The dominance and three-dimensional intertwined distribution of these organisms were confirmed by nonintrusive scanning microscopy. In contrast, the less pronounced biofilms were dominated by the heterocystous cyanobacterial genus Calothrix, two unicellular Entophysalis morphotypes, Lyngbya spp., and members of the Pseudanabaenaceae family. Cytophaga-Flavobacterium-Bacteroides and Alphaproteobacteria phylotypes were also retrieved from the beach rock. The microbial diversity of the dominant mat was accompanied by high nocturnal nitrogenase activities (as determined by in situ acetylene reduction assays). A new DGGE nifH gene optimization approach for cyanobacterial nitrogen fixers showed that the sequences retrieved from the dominant mat were related to nonheterocystous uncultured cyanobacterial phylotypes, only distantly related to sequences of nitrogen-fixing cultured cyanobacteria. These data stress the occurrence and importance of nonheterocystous epilithic cyanobacteria, and it is hypothesized that such epilithic cyanobacteria are the principal nitrogen fixers of the Heron Island beach rock. PMID- 17416690 TI - Binding of Cyt1Aa and Cry11Aa toxins of Bacillus thuringiensis serovar israelensis to brush border membrane vesicles of Tipula paludosa (Diptera: Nematocera) and subsequent pore formation. AB - Bacillus thuringiensis serovar israelensis (B. thuringiensis subsp. israelensis) produces four insecticidal crystal proteins (ICPs) (Cry4A, Cry4B, Cry11A, and Cyt1A). Toxicity of recombinant B. thuringiensis subsp. israelensis strains expressing only one of the toxins was determined with first instars of Tipula paludosa (Diptera: Nematocera). Cyt1A was the most toxic protein, whereas Cry4A, Cry4B, and Cry11A were virtually nontoxic. Synergistic effects were recorded when Cry4A and/or Cry4B was combined with Cyt1A but not with Cry11A. The binding and pore formation are key steps in the mode of action of B. thuringiensis subsp. israelensis ICPs. Binding and pore-forming activity of Cry11Aa, which is the most toxic protein against mosquitoes, and Cyt1Aa to brush border membrane vesicles (BBMVs) of T. paludosa were analyzed. Solubilization of Cry11Aa resulted in two fragments, with apparent molecular masses of 32 and 36 kDa. No binding of the 36 kDa fragment to T. paludosa BBMVs was detected, whereas the 32-kDa fragment bound to T. paludosa BBMVs. Only a partial reduction of binding of this fragment was observed in competition experiments, indicating a low specificity of the binding. In contrast to results for mosquitoes, the Cyt1Aa protein bound specifically to the BBMVs of T. paludosa, suggesting an insecticidal mechanism based on a receptor-mediated action, as described for Cry proteins. Cry11Aa and Cyt1Aa toxins were both able to produce pores in T. paludosa BBMVs. Protease treatment with trypsin and proteinase K, previously reported to activate Cry11Aa and Cyt1Aa toxins, respectively, had the opposite effect. A higher efficiency in pore formation was observed when Cyt1A was proteinase K treated, while the activity of trypsin-treated Cry11Aa was reduced. Results on binding and pore formation are consistent with results on ICP toxicity and synergistic effect with Cyt1Aa in T. paludosa. PMID- 17416689 TI - Phylogenetic composition of Rocky Mountain endolithic microbial ecosystems. AB - The endolithic environment, the pore space in rocks, is a ubiquitous microbial habitat. Photosynthesis-based endolithic communities inhabit the outer few millimeters to centimeters of rocks exposed to the surface. Such endolithic ecosystems have been proposed as simple, tractable models for understanding basic principles in microbial ecology. In order to test previously conceived hypotheses about endolithic ecosystems, we studied selected endolithic communities in the Rocky Mountain region of the United States with culture-independent molecular methods. Community compositions were determined by determining rRNA gene sequence contents, and communities were compared using statistical phylogenetic methods. The results indicate that endolithic ecosystems are seeded from a select, global metacommunity and form true ecological communities that are among the simplest microbial ecosystems known. Statistical analysis showed that biogeographical characteristics that control community composition, such as rock type, are more complex than predicted. Collectively, results of this study support the idea that patterns of microbial diversity found in endolithic communities are governed by principles similar to those observed in macroecological systems. PMID- 17416691 TI - Salivaricin P, one of a family of two-component antilisterial bacteriocins produced by intestinal isolates of Lactobacillus salivarius. AB - Lactobacillus salivarius DPC6005, a porcine intestinal isolate, produces a two component bacteriocin, salivaricin P, with homology to ABP-118 produced by a human probiotic L. salivarius strain. Indeed, molecular characterization revealed that while the peptides Sln1 and ABP-118alpha are identical, their companion peptides (Sln2 and ABP-118beta, respectively) differ by two amino acids. This observation suggests that two-component bacteriocins may be a common feature of intestinal L. salivarius strains. PMID- 17416692 TI - Microbial diversity in natural asphalts of the Rancho La Brea Tar Pits. AB - Bacteria commonly inhabit subsurface oil reservoirs, but almost nothing is known yet about microorganisms that live in naturally occurring terrestrial oil seeps and natural asphalts that are comprised of highly recalcitrant petroleum hydrocarbons. Here we report the first survey of microbial diversity in ca. 28,000-year-old samples of natural asphalts from the Rancho La Brea Tar Pits in Los Angeles, CA. Microbiological studies included analyses of 16S rRNA gene sequences and DNA encoding aromatic ring-hydroxylating dioxygenases from two tar pits differing in chemical composition. Our results revealed a wide range of phylogenetic groups within the Archaea and Bacteria domains, in which individual taxonomic clusters were comprised of sets of closely related species within novel genera and families. Fluorescent staining of asphalt-soil particles using phylogenetic probes for Archaea, Bacteria, and Pseudomonas showed coexistence of mixed microbial communities at high cell densities. Genes encoding dioxygenases included three novel clusters of enzymes. The discovery of life in the tar pits provides an avenue for further studies of the evolution of enzymes and catabolic pathways for bacteria that have been exposed to complex hydrocarbons for millennia. These bacteria also should have application for industrial microbiology and bioremediation. PMID- 17416693 TI - Detection and genotyping of Arcobacter and Campylobacter isolates from retail chicken samples by use of DNA oligonucleotide arrays. AB - To explore the use of DNA microarrays for pathogen detection in food, we produced DNA oligonucleotide arrays to simultaneously determine the presence of Arcobacter and the presence of Campylobacter in retail chicken samples. Probes were selected that target housekeeping and virulence-associated genes in both Arcobacter butzleri and thermotolerant Campylobacter jejuni and Campylobacter coli. These microarrays showed a high level of probe specificity; the signal intensities detected for A. butzleri, C. coli, or C. jejuni probes were at least 10-fold higher than the background levels. Specific identification of A. butzleri, C. coli, and C. jejuni was achieved without the need for a PCR amplification step. By adapting an isolation method that employed membrane filtration and selective media, C. jejuni isolates were recovered from package liquid from whole chicken carcasses prior to enrichment. Increasing the time of enrichment resulted in the isolation of A. butzleri and increased the recovery of C. jejuni. C. jejuni isolates were further classified by using an additional subset of probes targeting the lipooligosaccharide (LOS) biosynthesis locus. Our results demonstrated that most of the C. jejuni isolates likely possess class B, C, or H LOS. Validation experiments demonstrated that the DNA microarray had a detection sensitivity threshold of approximately 10,000 C. jejuni cells. Interestingly, the use of C. jejuni sequence-specific primers to label genomic DNA improved the sensitivity of this DNA microarray for detection of C. jejuni in whole chicken carcass samples. C. jejuni was efficiently detected directly both in package liquid from whole chicken carcasses and in enrichment broths. PMID- 17416694 TI - Novel methods for genetic transformation of natural Bacillus subtilis isolates used to study the regulation of the mycosubtilin and surfactin synthetases. AB - Natural isolates of Bacillus subtilis are often difficult to transform due to their low genetic competence levels. Here we describe two methods that stimulate natural transformation. The first method uses plasmid pGSP12, which expresses the competence transcription factor ComK and stimulates competence development about 100-fold. The second method stimulates Campbell-type recombination of DNA ligation mixtures in B. subtilis by the addition of polyethylene glycol. We employed these novel methods to study the regulation of the synthetases for the lipopeptide antibiotics mycosubtilin (myc) and surfactin (srfA) in B. subtilis strain ATCC 6633. By means of lacZ reporter fusions, it was shown that the expression of srfA is >100 times lower in strain ATCC 6633 than in the laboratory strain B. subtilis 168. Expression of the myc operon was highest in rich medium, whereas srfA expression reached maximal levels in minimal medium. Further genetic analyses showed that the srfA operon is mainly regulated by the response regulator ComA, while the myc operon is primarily regulated by the transition state regulator AbrB. Although there is in vitro evidence for a synergistic activity of mycosubtilin and surfactin, the expression of both lipopeptide antibiotics is clearly not coordinated. PMID- 17416695 TI - Effect of Phosphorus on survival of Escherichia coli in drinking water biofilms. AB - The effect of phosphorus addition on survival of Escherichia coli in an experimental drinking water distribution system was investigated. Higher phosphorus concentrations prolonged the survival of culturable E. coli in water and biofilms. Although phosphorus addition did not affect viable but not culturable (VBNC) E. coli in biofilms, these structures could act as a reservoir of VBNC forms of E. coli in drinking water distribution systems. PMID- 17416697 TI - Telling their stories, telling our stories: physicians' experiences with patients who decide to forgo or stop treatment for cancer. AB - There is currently very little research on how physicians respond to patients with cancer who decide to forgo or stop medically recommended "curative" therapy. The purpose of this article is to report on a qualitative study with 12 oncology specialists in Israel and Australia that addresses this question. The findings indicate that physicians tend to construct patients and their decisions in terms of mutually exclusive categories that focus on curability of the disease, rationality of the patient's decision, and patients' personal attributes. Physicians' constructions of their experience focus on uncertainty and concern. Although contextual factors play a role in how physicians act in this situation, Israeli and Australian oncologists are remarkably similar in how they describe their own and their patients' experiences. PMID- 17416698 TI - The implications of dying cancer patients' talk on cardiopulmonary resuscitation and do-not-resuscitate orders. AB - Current medical emphasis on autonomy requires that patients be primary in authorizing do-not-resuscitate (DNR) orders, countermanding provision of cardiopulmonary resuscitation (CPR) on terminally ill patients. The assumptions that patients make regarding CPR and DNR orders will influence their choices about them. Using discursive analysis, the authors examined the speech of 28 patients dying of cancer regarding the appropriateness of refraining from CPR or of instituting DNR orders. Most participants identified CPR as inappropriate in their circumstances, favoring institution of DNR orders. However, a minority drew on dominant construals of DNR orders and CPR to locate themselves outside the category of suitable candidates for DNR orders, thus justifying a preference for CPR--even though some had current DNR orders. Doctors' and patients' assessments of eligibility for DNR orders might not coincide, and when patient autonomy is presumed by patients to be determinant, discrepancies between patient expectations and instituted medical practice are inevitable. PMID- 17416699 TI - Family photographs: expressions of parents raising children with disabilities. AB - The purpose of this qualitative, descriptive study was to capture the everyday lives of parents raising a child with a disability. Fifteen parental dyads raising a child with Down syndrome, developmental disabilities, visual impairments, or speech disorders used a disposable camera to photograph images important to them. Interviews about these photographs reflected everyday life. Four themes emerged from content analysis of the photos: active activities, quiet activities, activities of daily living, and care of the child with a disability. Six themes emerged when discussing photographs participants would have liked to take but did not. These themes were acceptance, concern for the future, joys, struggles, balance of good and bad times, and normalcy. Findings support the usefulness of photography as a method of obtaining information from parents raising a child with a disability through qualitative means and provide nurses with a beginning understanding of these parents' lives. PMID- 17416700 TI - Multilingual translation issues in qualitative research: reflections on a metaphorical process. AB - Focus on the translation and validation of measurement instruments has left a gap in the discussion on how to construct multilingual qualitative tools, such as interviews. Traditional methods of forward and backward translation have been criticized for weak conceptual equivalence, a crucial issue when multiple language interview methods are used. Through a creative arts metaphor of weaving, the authors describe an alternative process of multicentric translation used in the development of an interview guide designed to explore the impact of transition on palliative care patients in six European countries. Four identified core constructs illuminate this multicentric process: Cohesion, Congruence, Clarity, and Courtesy. Mutual reciprocity between researcher and translator offers greater possibility for construction of nuance and meaning, particularly where cultural parameters influence the collection and meaning of sensitive data from vulnerable populations. The translator therefore becomes a collaborator in the research process, which strengthens the rigor of language-based inquiry. PMID- 17416701 TI - Gender and the gynecological examination: women's identities in doctors' narratives. AB - The authors explore the constructions of gender in male doctors' narratives of gynecological examinations. Focusing on the ways in which gender identities are constructed in the stories of the medical encounter, they argue, first, that gender is more flexible during the visit with a gynecologist than has been suggested. Gendered identities are assumed and put aside as the interaction progresses, with its final stage--the pelvic examination--being constructed with gender removed. Second, they argue that undressing is invested with a special status during the examination. It is a gendered rite of passage between the two different ungendered subject positions of the doctor and the patient. They conclude that contrary to the assumptions in the literature on gynecological interactions, it is the genderization of undressing that is most conducive to securing the least face-threatening gynecological examination for the woman. PMID- 17416702 TI - Stepping across the line: information sharing, truth telling, and the role of the personal carer in the Australian nursing home. AB - The author draws on an Australian study using multiple qualitative methods to investigate truth telling in aged care. Thematic analysis of data from five nursing homes involving 23 personal care assistants revealed participants' role understanding as influencing their perceptions about truth telling in practice. Five themes emerged: role as the happy comfort carer, division of labor, division of disclosure, role tension and frustration, and managing the division of disclosure. Role emphasis on comfort and happiness and a dominant perception that telling the truth can cause harm mean that disclosure will be withheld, edited, or partial. Participants'role understanding divides labor and disclosure responsibility between the personal carer and registered nurse. Personal carers' strategies for managing the division of disclosure include game playing, obfuscation, lying (denial), and the use of nonverbals. These perceptions about personal carer role, information sharing, and truth telling are paramount for understanding and improving nursing home eldercare. PMID- 17416703 TI - Lost in translation, or the true text: mental health nursing representations of psychology. AB - One theme of academic discourse and research in mental health nursing is the exploration and application of psychosocial models of nursing practice. Despite this, the influence of disciplinary psychological knowledge on practitioners' talk about nursing practice has not been extensively researched. To address this gap, the authors analyzed talk about psychological work using transcripts of 10 focus groups involving 59 mental health nurses. Nurses identified a psychological domain of practice as central to their work. Given the amount of time spent with clients, nurses are the prime resource for psychological work. Psychological talk was organized into three categories related to the nursing process and organizational context and analyzed through empowerment and critical perspectives on power. Although technical ("formal") and everyday ("informal") discourses were generally well integrated, the authors question the oral basis to this body of knowledge in terms of accountability to service users and as a marker of disempowerment. PMID- 17416704 TI - Narrative accounts of hereditary risk: knowledge about family history, lay theories of disease, and "internal" and "external" causation. AB - In this study, the authors sought to examine how risk information is articulated in relation to health problems that people identify as personally important and relevant. The respondents were receptive to health education messages, using different types of information in relation to its personal relevance and as a resource for managing and exercising control over perceived risk. People were not fatalistic about disease risk, as reported in previous research. Instead, they were responsive to complex public health messages and actively engaged in rationalizing their health risks, although this did not necessarily result in behavioral change. Consequently, a theoretical distinction exists between taking responsibility for evaluating complex public health messages and taking responsibility for behavioral change. The authors conclude that people's rationalizations about health risks often mirror the medical model of disease, suggesting that they are responsive to, and not fatalistic toward, such public health information. PMID- 17416705 TI - Applications of performance ethnography in nursing. AB - In this article, the authors explore the origins of and theory behind performance ethnography and demonstrate how it can be used effectively in nursing research, education, and practice. They have included a performance text created by one of the authors based on semistructured, qualitative interviews with parents of children with genetic conditions. The text highlights the lived experiences of the parents, allowing the "audience" to appreciate more fully the complexities of parenting children with genetic conditions. The authors also discuss specific ways in which nurses can use performance ethnography in their professional practice, including several examples. Performance ethnography can be a creative method for nurses to communicate with patients and families, research audiences, colleagues, and students. Clinicians, researchers, educators, and students can learn to use performance ethnography to achieve the goal of a clear understanding of human needs and conditions for improved quality of care. PMID- 17416706 TI - Double vision uncertainty: the bilingual researcher and the ethics of cross language research. AB - Translation is a significant factor in cross-language research and can make an overarching impact on research outcomes and ethical adequacy. In this article, the author examines ethical considerations of language translation and interpretation in human science research. The focus is narrowed to ethical aspects of research design in which a bilingual researcher assumes a double role, as an interpreter and translator in his or her inquiry with monolingual, non English-speaking research participants. The bilingual researcher has a great advantage of expertise and clear vision in the cross-cultural ethical environment. However, such clarity of the ethical vision can cause many reasonable doubts, because it might not fully fit into the framework of standardized, rational ethics. The author shows that the dualism of the bilingual researcher's position can either induce caution or carry solutions, but both aspects of this role contribute to its essential significance for good cross cultural knowledge exchange. PMID- 17416707 TI - Doing synchronous online focus groups with young people: methodological reflections. AB - Although online focus groups are emerging as a worthwhile methodological approach for qualitative researchers, reporting has been constrained in several ways. The majority of studies report asynchronous groups, whereas others employ synchronous exchanges, the efficacy of which with young people has seldom been explored. Considering the popularity of the Internet as a communication tool for young people, this missed opportunity is surprising. Based on a series of synchronous online focus groups with young people, the authors explore why this approach might be an effective way of engaging young people with appearance-related concerns in research. In this article, they discuss the process of hosting and moderating synchronous online focus groups, highlighting some of the ethical, pragmatic, and personal challenges that might face researchers using this method. Through a reflexive approach, they intend to inform and encourage qualitative researchers to consider alternative ways of engaging young people in research. PMID- 17416708 TI - The inevitable pull of the river's current: interpretations derived from a single text using multiple research traditions. AB - The purpose of this article is to make visible the multiple ways in which doctoral students from various qualitative research traditions learned to think, read, and write interpretively as they completed an assignment requiring the interpretive analysis of a common interview transcript. Students were asked to offer a convincing account of the text and to demonstrate an understanding of what it means to interpret within their selected research tradition. Shared and disputed meanings arising from the interpretive process are presented and discussed. This description of their collective experience might be useful to novice researchers and their mentors. PMID- 17416709 TI - A tribute to Agnes Marie Aamodt, PhD, RN, FAAN, 1921-2006. PMID- 17416710 TI - American Academy of Nursing Expert Panel report: developing cultural competence to eliminate health disparities in ethnic minorities and other vulnerable populations. AB - The members of the Expert Panel on Cultural Competence of the American Academy of Nursing (AAN) envisioned this article to serve as a catalyst to action by the Academy to take the lead in ensuring that measurable outcomes be achieved that reduce or eliminate health disparities commonly found among racial, ethnic, uninsured, underserved, and underrepresented populations residing throughout the United States. The purposes of this article are to (a) assess current issues related to closing the gap in health disparities and achieving cultural competence, (b) discuss a beginning plan of action from the Expert Panel on Cultural Competence for future endeavors and continued work in these areas beyond the 2002 annual conference on Closing the Gap in Health Disparities, and (c) provide clearly delineated recommendations to assist the Academy to plan strategies and to step forward in taking the lead in reshaping health care policies to eliminate health care and health disparities. PMID- 17416711 TI - Culturally congruent care: putting the puzzle together. AB - This article presents the 3-D puzzle model of culturally congruent care, defines the levels and constructs employed by this model, presents some assumptions, and lays out some basic propositions as a foundation for further work. There are many extant frameworks and theories of culture and cultural competence in nursing and health care; the model presented here draws heavily from prior work and is an attempt to present a synthesis of concepts and processes in a new way. The ideas presented here build particularly on pioneering work in transcultural nursing by Leininger. Leininger's work specifically focuses on the use of qualitative methods to understand the ways in which culture influences nursing care from an emic, or insider, perspective. The 3-D puzzle model extends Leininger's work to include concrete articulations of constructs relevant to design and implementation of intervention strategies for teaching and measuring competency among nurses and other providers. PMID- 17416712 TI - Validating the General Sleep Disturbance Scale among Chinese American parents with hospitalized infants. AB - The purpose of this cross-sectional descriptive study was to assess the reliability and validity of the English and Chinese versions of the General Sleep Disturbance Scale (GSDS) among Chinese American parents with hospitalized infants. A convenience sample of 22 mothers and 22 fathers in the San Francisco area participated in this study. Cronbach alpha coefficients were .85 and .81 for the English and Chinese versions, respectively. Concurrent validity was demonstrated through correlations with wrist actigraphy readings. Although results of the study indicate that the GSDS is an acceptable instrument to measure parents' sleep disturbance among the Chinese American population, a need for further work was identified. PMID- 17416713 TI - The concepts of health and preventive health practices of Chinese Australian women in relation to cancer screening. AB - Despite an emphasis on mammographic screening in Australia, Chinese Australian women have low participation rates. This qualitative study investigated how concepts of health and health promotion influence Chinese Australian women's decisions to participate in cancer screening, which is an important issue for nurses who work with multicultural populations. In-depth interviews were conducted with 20 Chinese Australian women. Using thematic data analysis, the findings showed that health and illness are taken-for-granted experiences of everyday life. When they were asymptomatic, most informants saw no reason to suspect that they may have diseases. Consistent with these health beliefs, the women focused on preserving and promoting health and overall well-being in everyday life rather than attempting to detect hidden disease by screening. These ideas and practices influenced behavior in relation to cancer diagnosis and in particular toward mammography. PMID- 17416714 TI - Modification and translation of the Thai version of the Youth Risk Behavior Survey. AB - The Youth Risk Behavior Survey (YRBS) has been used as a standard and updated as required for 15 years to measure risk behavior among American youth. This article discusses the modification and translation of the YRBS to be used to measure risk among youth in Thailand. The steps involved included not only modification and translation but also testing of cross-cultural equivalences (semantic, content, and conceptual) and assessment of feasibility and acceptability of the modified survey among adolescents in Thailand. The process is described and findings discussed. PMID- 17416715 TI - It's a long way from the office to the creek bed: remote area mental health nursing in Australia. AB - Mental health nursing in remote Australian Aboriginal communities provides opportunities and challenges unique from other experiences of nursing. The purpose of this article was to explore how mental health nurses experience working in remote communities and how they developed relevant knowledge and skills. Six remote area mental health nurses participated in audiotaped conversational style interviews. Narratives were transcribed and analyzed thematically. Themes identified were being focused on relationships, adapting to the culture, struggling with two contexts, being enraptured by place, and being transformed. The participants adapted and developed ways of working at the interface of another culture and also derived personal and professional benefit from the experience. PMID- 17416716 TI - From their own voices: the lived experience of African American registered nurses. AB - This phenomenological study described the lived experience of African American registered nurses providing nursing care to individuals, families, and communities in southeast Louisiana. Data were collected from 13 African American registered nurses using semistructured interviews and a focus group. Analysis of the phenomenological data revealed two essential themes, (a) connecting with the patient and (b) proving yourself; and four incidental themes, (a) a fulfilling dream, (b) being invisible and voiceless, (c) surviving and persevering, and (d) mentoring and role modeling. The findings revealed that the general perception among participants was that they were not fully accepted as equal professionals by their Caucasian nurse colleagues, other health care providers, and sometimes patients. The findings of the study indicate the immediate need to address and resolve the issues of diversity within the nursing profession. Nursing will also have to reform its system and practices to embrace and support diversity. PMID- 17416717 TI - Breaching the wall: interviewing people from other cultures. AB - The interview as a data collection tool is an essential component of qualitative research. Many nurses are familiar with the process of interviewing through its use in the practice environment; however, in-depth interviewing for the purpose of research is a unique process. The ability to conduct an effective in-depth interview requires skill in the use of specific techniques, in particular when interviewing people from other cultures. A number of factors specific to the researcher, the participant, and the research context can affect the interview procedure. As global margins diminish, nurse researchers will increasingly find themselves working with people from ethnic groups that are different from the dominant culture. This article discusses strategies to improve the interview process in such circumstances. Techniques to enhance the process, along with avoidable potential pitfalls, will be illustrated using an example of conducting research with participants from the culturally diverse environment of Malaysian Borneo. PMID- 17416718 TI - Culturally competent interventions for Hispanic adults with type 2 diabetes: a systematic review. AB - Culturally competent interventions have been developed to improve outcomes for Hispanic adults with type 2 diabetes. The purpose of this systematic review is to synthesize the research on culturally competent interventions for this vulnerable population. A systematic approach was used to locate empirical reports (n = 11). Interventions were multifaceted with the majority demonstrating significant improvements in clinical outcomes, behavioral outcomes, and diabetes-related knowledge. Culturally competent interventions have the potential to improve outcomes in Hispanic adults with type 2 diabetes. However, improvements were modest and attrition was moderate to high in many studies. Addressing linguistic and cultural barriers to care are important beginnings to improving health outcomes for Hispanic adults with type 2 diabetes. PMID- 17416719 TI - Being in another world: transcultural student experiences using service learning with families who are homeless. AB - Developing skills in cultural competence is a recognized theoretical strategy in schools of nursing. Nursing faculty know that students need to be sensitized to the concept of diversity; however, many are struggling with the best way to teach cultural competence. This article describes transcultural experiences from service learning clinical rotations at a family homeless shelter, described by students as being in another world. Student narratives provide valuable information about structuring clinical learning activities to promote understanding of cultural differences and similarities. Clinical experiences using a traditional model versus those using service learning, the role of reflection, and teaching strategies promoting transcultural learning through service learning are explored. PMID- 17416720 TI - Mental health issues among urban Korean American immigrants. AB - Korean immigrants' perception and understanding of mental health and illness were surveyed during four monthly mental health seminars in New York City. In all, 86 Korean immigrants attended the seminars and 34 completed the survey. The seminar participants were primarily financially stable Korean women who were married, educated, and had lived in America more than 10 years. All seminar leaders were Korean immigrants who were working in the mental health field and/or educational setting. Most of the participants acknowledged the need for mental health services but did not seek professional help and coped with the stressors of immigrant life by endurance, patience, and religion. Feedback from seminar leaders noted the following: (a) greater seminar attendance than anticipated, (b) participants' openness to their mental illness issues, (c) need for tailored mental health program for Koreans. Findings support an understanding of the Korean immigrants' mental health issues as complex, chronic, and serious. PMID- 17416721 TI - President's message: moving forward, making a difference. PMID- 17416722 TI - Complications and long-term outcomes of the ligation of congenital portosystemic shunts in 49 cats. AB - Only two of 49 cats undergoing surgical ligation of congenital extra- and intrahepatic portosystemic shunts died perioperatively, a mortality rate comparable with the mortality rates of dogs undergoing surgical attenuation of congenital portosystemic shunts and cats in which the shunts are attenuated with an ameroid ring constrictor. Thirty (83 per cent) of the 36 cats for which long term information was available were still alive at a median follow-up period of 47 months (range six to 105 months); the outcome was excellent (no clinical signs) in 20 of them (median follow-up 37 months, range six to 105 months) and good (minimal clinical signs) in seven (median follow-up 39 months, range 10 to 73 months) and none of these 27 cats was on any long-term medication or special diet. The only major cause of morbidity was the development of neurological signs in 18 (37 per cent) of the cats. These included seizures and a wide variety of other neurological signs, and their development and persistence was not affected by the presence of preoperative seizures, the type of shunt, the degree of shunt attenuation or the age of the cat. The serum concentrations of ammonia and preprandial bile acids were normal or significantly below normal in the cats with neurological signs. Liver histopathology was similar in the cats with and without neurological signs. Ten (56 per cent) of the 18 cats that developed neurological signs recovered normal neurological function long term. PMID- 17416723 TI - Effects of housing and intake of methionine on the growth and wear of hoof horn and the conformation of the hooves of first-lactation Holstein heifers. AB - This study investigated the effects of housing primiparous lactating Holstein heifers either in cubicles with butyl rubber mats, cubicles with thicker mattresses filled with chopped rubber, or straw yards, and supplementing them with methionine for the first 13 weeks of lactation on the rates of growth of their hoof horn and the conformation of their feet. Overall, the rate of growth of the hoof horn increased significantly after calving but the rate of wear did not increase. In contrast, the type of housing had no significant effect on the growth of the hoof horn, but the heifers in straw yards had significantly lower rates of lateral horn wear. The changes in growth and wear rates resulted in changes in hoof conformation, including the toe angle and the width of the bearing surface, which suggested that heel bulb hypertrophy occurred after the heifers had been moved from soft underfoot conditions on to concrete. Providing 115 per cent of their estimated methionine requirements had no significant effect on the rates of growth and wear of the hoof horn. PMID- 17416724 TI - Preliminary studies of particle-mediated gene delivery to the joints of dogs. AB - This paper describes a preliminary evaluation of particle-mediated bombardment via the Helios gene gun for the delivery of therapeutic genes to synovial cells in culture. A reporter gene, enhanced green fluorescent protein, was delivered to rabbit synovial fibroblasts (HIG-82) using gold particle (1.0 microm) bombardment to evaluate transfection efficiency at helium pressures of 100 and 150 psi. Transfection of cells occurred at these pressures despite some cell death. The in vitro delivery of gold particles to samples of synovial membrane and articular cartilage from a freshly euthanased dog was also studied to examine depth of penetration of gold particles (1.0 microm) at helium pressures of 250 and 500 psi. Light microscopical examination of histological sections of the synovial membrane showed that particles of gold had penetrated the lining cells of the synovium. However, no gold particles had penetrated the articular cartilage even at 500 psi. PMID- 17416725 TI - Outbreak of Newcastle disease in pheasants (Phasianus colchicus) in south-east England in July 2005. PMID- 17416726 TI - Isolation of Mycoplasma bovis from the respiratory tract of cattle in Bosnia and Herzegovina. PMID- 17416727 TI - Efficacy of levamisole and Tarantula cubensis venom for the treatment of bovine cutaneous papillomatosis. PMID- 17416728 TI - Controlling sheep scab by eradication. PMID- 17416730 TI - The DELLA domain of GA INSENSITIVE mediates the interaction with the GA INSENSITIVE DWARF1A gibberellin receptor of Arabidopsis. AB - Gibberellic acid (GA) promotes seed germination, elongation growth, and flowering time in plants. GA responses are repressed by DELLA proteins, which contain an N terminal DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. Mutations of or within the DELLA domain of DELLA repressors have been described for species including Arabidopsis thaliana, wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare), and we show that these mutations confer GA insensitivity when introduced into the Arabidopsis GA INSENSITIVE (GAI) DELLA repressor. We also demonstrate that Arabidopsis mutants lacking the three GA INSENSITIVE DWARF1 (GID1) GA receptor genes are GA insensitive with respect to GA-promoted growth responses, GA-promoted DELLA repressor degradation, and GA regulated gene expression. Our genetic interaction studies indicate that GAI and its close homolog REPRESSOR OF ga1-3 are the major growth repressors in a GA receptor mutant background. We further demonstrate that the GA insensitivity of the GAI DELLA domain mutants is explained in all cases by the inability of the mutant proteins to interact with the GID1A GA receptor. Since we found that the GAI DELLA domain alone can mediate GA-dependent GID1A interactions, we propose that the DELLA domain functions as a receiver domain for activated GA receptors. PMID- 17416731 TI - Replication-independent long-distance trafficking by viral RNAs in Nicotiana benthamiana. AB - Viruses with separately encapsidated genomes could have their genomes introduced into different leaves of a plant, thus necessitating long-distance trafficking of the viral RNAs for successful infection. To examine this possibility, individual or combinations of genome segments from the tripartite Brome mosaic virus (BMV) were transiently expressed in leaves of Nicotiana benthamiana plants using engineered Agrobacterium tumefaciens. BMV RNA3 was found to traffic from the initial site of expression to other leaves of the plant, as detected by RNA gel blot analyses and also by the expression of an endoplasmic reticulum-targeted green fluorescent protein. When RNA3 trafficked into leaves containing the BMV replication enzymes, RNA replication, transcription, and virion production were observed. RNA3 trafficking occurred even when it did not encode the movement or capsid proteins. However, coexpression of the movement protein increased the trafficking of BMV RNAs. BMV RNA1 and RNA2 could also traffic throughout the plant, but less efficiently than RNA3. All three BMV RNAs trafficked bidirectionally to sink leaves near the apical meristem as well as to the source leaves at the bottom of the stem, suggesting that trafficking used the phloem. These results demonstrate that BMV RNAs can use a replication-independent mechanism to traffic in N. benthamiana. PMID- 17416732 TI - SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis. AB - SIZ1 is a SUMO E3 ligase that facilitates conjugation of SUMO to protein substrates. siz1-2 and siz1-3 T-DNA insertion alleles that caused freezing and chilling sensitivities were complemented genetically by expressing SIZ1, indicating that the SIZ1 is a controller of low temperature adaptation in plants. Cold-induced expression of CBF/DREB1, particularly of CBF3/DREB1A, and of the regulon genes was repressed by siz1. siz1 did not affect expression of ICE1, which encodes a MYC transcription factor that is a controller of CBF3/DREB1A. A K393R substitution in ICE1 [ICE1(K393R)] blocked SIZ1-mediated sumoylation in vitro and in protoplasts identifying the K393 residue as the principal site of SUMO conjugation. SIZ1-dependent sumoylation of ICE1 in protoplasts was moderately induced by cold. Sumoylation of recombinant ICE1 reduced polyubiquitination of the protein in vitro. ICE1(K393R) expression in wild-type plants repressed cold-induced CBF3/DREB1A expression and increased freezing sensitivity. Furthermore, expression of ICE1(K393R) induced transcript accumulation of MYB15, which encodes a MYB transcription factor that is a negative regulator of CBF/DREB1. SIZ1-dependent sumoylation of ICE1 may activate and/or stabilize the protein, facilitating expression of CBF3/DREB1A and repression of MYB15, leading to low temperature tolerance. PMID- 17416733 TI - Rice NON-YELLOW COLORING1 is involved in light-harvesting complex II and grana degradation during leaf senescence. AB - Chlorophyll degradation is an aspect of leaf senescence, which is an active process to salvage nutrients from old tissues. non-yellow coloring1 (nyc1) is a rice (Oryza sativa) stay-green mutant in which chlorophyll degradation during senescence is impaired. Pigment analysis revealed that degradation of not only chlorophylls but also light-harvesting complex II (LHCII)-bound carotenoids was repressed in nyc1, in which most LHCII isoforms were selectively retained during senescence. Ultrastructural analysis of nyc1 chloroplasts revealed that large and thick grana were present even in the late stage of senescence, suggesting that degradation of LHCII is required for the proper degeneration of thylakoid membranes. Map-based cloning of NYC1 revealed that it encodes a chloroplast localized short-chain dehydrogenase/reductase (SDR) with three transmembrane domains. The predicted structure of the NYC1 protein and the phenotype of the nyc1 mutant suggest the possibility that NYC1 is a chlorophyll b reductase. Although we were unable to detect the chlorophyll b reductase activity of NYC1, NOL (for NYC1-like), a protein closely related to NYC1 in rice, showed chlorophyll b reductase activity in vitro. We suggest that NYC1 and NOL encode chlorophyll b reductases with divergent functions. Our data collectively suggest that the identified SDR protein NYC1 plays essential roles in the regulation of LHCII and thylakoid membrane degradation during senescence. PMID- 17416734 TI - The balance between protein synthesis and degradation in chloroplasts determines leaf variegation in Arabidopsis yellow variegated mutants. AB - An Arabidopsis thaliana leaf-variegated mutant yellow variegated2 (var2) results from loss of FtsH2, a major component of the chloroplast FtsH complex. FtsH is an ATP-dependent metalloprotease in thylakoid membranes and degrades several chloroplastic proteins. To understand the role of proteolysis by FtsH and mechanisms leading to leaf variegation, we characterized the second-site recessive mutation fu-gaeri1 (fug1) that suppressed leaf variegation of var2. Map based cloning and subsequent characterization of the FUG1 locus demonstrated that it encodes a protein homologous to prokaryotic translation initiation factor 2 (cpIF2) located in chloroplasts. We show evidence that cpIF2 indeed functions in chloroplast protein synthesis in vivo. Suppression of leaf variegation by fug1 is observed not only in var2 but also in var1 (lacking FtsH5) and var1 var2. Thus, suppression of leaf variegation caused by loss of FtsHs is most likely attributed to reduced protein synthesis in chloroplasts. This hypothesis was further supported by the observation that another viable mutation in chloroplast translation elongation factor G also suppresses leaf variegation in var2. We propose that the balance between protein synthesis and degradation is one of the determining factors leading to the variegated phenotype in Arabidopsis leaves. PMID- 17416735 TI - High serum-free light chain levels and their rapid reduction in response to therapy define an aggressive multiple myeloma subtype with poor prognosis. AB - Serum-free light chain (SFLC) levels are useful for diagnosing nonsecretory myeloma and monitoring response in light-chain-only disease, especially in the presence of renal failure. As part of a tandem autotransplantation trial for newly diagnosed multiple myeloma, SFLC levels were measured at baseline, within 7 days of starting the first cycle, and before both the second induction cycle and the first transplantation. SFLC baseline levels higher than 75 mg/dL (top tertile) identified 33% of 301 patients with higher near-complete response rate (n-CR) to induction therapy (37% vs 20%, P = .002) yet inferior 24-month overall survival (OS: 76% vs 91%, P < .001) and event-free survival (EFS: 73% vs 90%, P < .001), retaining independent prognostic significance for both EFS (HR = 2.40, P = .008) and OS (HR = 2.43, P = .016). Baseline SFLC higher than 75 mg/dL was associated with light-chain-only secretion (P < .001), creatinine level 176.8 microM (2 mg/dL) or higher (P < .001), beta-2-microglobulin 297.5 nM/L (3.5 mg/L) or higher (P < .001), lactate dehydrogenase 190 U/L or higher (P < .001), and bone marrow plasmacytosis higher than 30% (P = .003). Additional independent adverse implications were conferred by top-tertile SFLC reductions before cycle 2 (OS: HR = 2.97, P = .003; EFS: HR = 2.56, P = .003) and before transplantation (OS: HR = 3.31, P = .001; EFS: HR = 2.65, P = .003). Unlike baseline and follow up analyses of serum and urine M-proteins, high SFLC levels at baseline reflecting more aggressive disease-and steeper reductions after therapy identified patients with inferior survival. PMID- 17416736 TI - NPM-ALK oncogenic kinase promotes cell-cycle progression through activation of JNK/cJun signaling in anaplastic large-cell lymphoma. AB - Anaplastic large-cell lymphoma (ALCL) frequently carries the t(2;5)(p23;q35), resulting in aberrant expression of nucleophosmin-anaplastic lymphoma kinase (NPM ALK). We show that in 293T and Jurkat cells, forced expression of active NPM-ALK, but not kinase-dead mutant NPM-ALK (210K>R), induced JNK and cJun phosphorylation, and this was linked to a dramatic increase in AP-1 transcriptional activity. Conversely, inhibition of ALK activity in NPM-ALK(+) ALCL cells resulted in a concentration-dependent dephosphorylation of JNK and cJun and decreased AP-1 DNA-binding. In addition, JNK physically binds NPM-ALK and is highly activated in cultured and primary NPM-ALK(+) ALCL cells. cJun phosphorylation in NPM-ALK(+) ALCL cells is mediated by JNKs, as shown by selective knocking down of JNK1 and JNK2 genes using siRNA. Inhibition of JNK activity using SP600125 decreased cJun phosphorylation and AP-1 transcriptional activity and this was associated with decreased cell proliferation and G2/M cell cycle arrest in a dose-dependent manner. Silencing of the cJun gene by siRNA led to a decreased S-phase cell-cycle fraction associated with upregulation of p21 and downregulation of cyclin D3 and cyclin A. Taken together, these findings reveal a novel function of NPM-ALK, phosphorylation and activation of JNK and cJun, which may contribute to uncontrolled cell-cycle progression and oncogenesis. PMID- 17416737 TI - The chemokine GRObeta mobilizes early hematopoietic stem cells characterized by enhanced homing and engraftment. AB - Mobilized peripheral blood hematopoietic stem cells (PBSCs) demonstrate accelerated engraftment compared with bone marrow; however, mechanisms responsible for enhanced engraftment remain unknown. PBSCs mobilized by GRObeta (GRObeta(Delta4)/CXCL2(Delta4)) or the combination of GRObeta(Delta4) plus granulocyte colony-stimulating factor (G-CSF) restore neutrophil and platelet recovery faster than G-CSF-mobilized PBSCs. To determine mechanisms responsible for faster hematopoietic recovery, we characterized immunophenotype and function of the GRObeta-mobilized grafts. PBSCs mobilized by GRObeta(Delta4) alone or with G-CSF contained significantly more Sca-1(+)-c-kit(+)-lineage(-) (SKL) cells and more primitive CD34(-)-SKL cells compared with cells mobilized by G-CSF and demonstrated superior competitive long-term repopulation activity, which continued to increase in secondary and tertiary recipients. GRObeta(Delta4) mobilized SKL cells adhered better to VCAM-1(+) endothelial cells compared with G CSF-mobilized cells. GRObeta(Delta4)-mobilized PBSCs did not migrate well to the chemokine stromal derived factor (SDF)-1alpha in vitro that was associated with higher CD26 expression. However, GRObeta(Delta4)-mobilized SKL and c-Kit(+) lineage(-) (KL) cells homed more efficiently to marrow in vivo, which was not affected by selective CXCR4 and CD26 antagonists. These data suggest that GRObeta(Delta4)-mobilized PBSCs are superior in reconstituting long-term hematopoiesis, which results from differential mobilization of early stem cells with enhanced homing and long-term repopulating capacity. In addition, homing and engraftment of GRObeta(Delta4)-mobilized cells is less dependent on the SDF 1alpha/CXCR4 axis. PMID- 17416738 TI - Similar but not the same: normobaric and hyperbaric pulmonary oxygen toxicity, the role of nitric oxide. AB - Pulmonary manifestations of oxygen toxicity were studied and quantified in rats breathing >98% O(2) at 1, 1.5, 2, 2.5, and 3 ATA to test our hypothesis that different patterns of pulmonary injury would emerge, reflecting a role for central nervous system (CNS) excitation by hyperbaric oxygen. At 1.5 atmosphere absolute (ATA) and below, the well-recognized pattern of diffuse pulmonary damage developed slowly with an extensive inflammatory response and destruction of the alveolar-capillary barrier leading to edema, impaired gas exchange, respiratory failure, and death; the severity of these effects increased with time over the 56 h period of observation. At higher inspired O(2) pressures, 2-3 ATA, pulmonary injury was greatly accelerated but less inflammatory in character, and events in the brain were a prelude to a distinct lung pathology. The CNS-mediated component of this lung injury could be attenuated by selective inhibition of neuronal nitric oxide synthase (nNOS) or by unilateral transection of the vagus nerve. We propose that extrapulmonary, neurogenic events predominate in the pathogenesis of acute pulmonary oxygen toxicity in hyperbaric oxygenation, as nNOS activity drives lung injury by modulating the output of central autonomic pathways. PMID- 17416739 TI - Prevention of endotoxin-induced systemic response by bone marrow-derived mesenchymal stem cells in mice. AB - Bone marrow-derived mesenchymal stem cells (BMDMSCs) appear to be important in repair of the chronic lung injury caused by bleomycin in mice. To determine effects of these BMDMSCs on an acute inflammatory response, we injected C57BL/6 mice intraperitoneally with 1 mg/kg endotoxin followed either by intravenous infusion of 5 x 10(5) BMDMSCs, the same number of lung fibroblasts, or an equal volume of normal saline solution. Lungs harvested 6, 24, and 48 h and 14 days after endotoxin showed that BMDMSC administration prevented endotoxin-induced lung inflammation, injury, and edema. Although we were able to detect donor cells in the lungs at 1 day after endotoxin, by 14 days no donor cells were detected. BMDMSC administration suppressed the endotoxin-induced increase in circulating proinflammatory cytokines without decreasing circulating levels of anti inflammatory mediators. Ex vivo cocultures of BMDMSC and lung cells from endotoxemic animals demonstrated a bilateral conversation in which lung cells stimulated proliferation and migration of stem cells and suppressed proinflammatory cytokine production by lung cells. We conclude that BMDMSCs decrease both the systemic and local inflammatory responses induced by endotoxin. These effects do not require either lung engraftment or differentiation of the stem cells and are due at least in part to the production of stem cell chemoattractants by the lungs and to humoral and physical interactions between stem cells and lung cells. We speculate that mobilization of this population of BMDMSCs may be a general mechanism for modulating an acute inflammatory response. PMID- 17416740 TI - Proinflammatory response of alveolar epithelial cells is enhanced by alveolar macrophage-produced TNF-alpha during pulmonary ischemia-reperfusion injury. AB - Pulmonary ischemia-reperfusion (IR) injury entails acute activation of alveolar macrophages followed by neutrophil sequestration. Although proinflammatory cytokines and chemokines such as TNF-alpha and monocyte chemoattractant protein-1 (MCP-1) from macrophages are known to modulate acute IR injury, the contribution of alveolar epithelial cells to IR injury and their intercellular interactions with other cell types such as alveolar macrophages and neutrophils remain unclear. In this study, we tested the hypothesis that following IR, alveolar macrophage-produced TNF-alpha further induces alveolar epithelial cells to produce key chemokines that could then contribute to subsequent lung injury through the recruitment of neutrophils. Cultured RAW264.7 macrophages and MLE-12 alveolar epithelial cells were subjected to acute hypoxia-reoxygenation (H/R) as an in vitro model of pulmonary IR. H/R (3 h/1 h) significantly induced KC, MCP-1, macrophage inflammatory protein-2 (MIP-2), RANTES, and IL-6 (but not TNF-alpha) by MLE-12 cells, whereas H/R induced TNF-alpha, MCP-1, RANTES, MIP-1alpha, and MIP-2 (but not KC) by RAW264.7 cells. These results were confirmed using primary murine alveolar macrophages and primary alveolar type II cells. Importantly, using macrophage and epithelial coculture methods, the specific production of TNF alpha by H/R-exposed RAW264.7 cells significantly induced proinflammatory cytokine/chemokine expression (KC, MCP-1, MIP-2, RANTES, and IL-6) by MLE-12 cells. Collectively, these results demonstrate that alveolar type II cells, in conjunction with alveolar macrophage-produced TNF-alpha, contribute to the initiation of acute pulmonary IR injury via a proinflammatory cascade. The release of key chemokines, such as KC and MIP-2, by activated type II cells may thus significantly contribute to neutrophil sequestration during IR injury. PMID- 17416741 TI - Behavioral cross-tolerance between repeated intracerebellar nicotine and acute Delta(9)-tetrahydrocannabinol-induced cerebellar ataxia: role of cerebellar nitric oxide. AB - We have previously demonstrated that acute intracerebellar nicotine or N-methyl-4 (3-pyridinyl)-3-buten-1-amine (RJR-2403), a selective alpha(4)beta(2) nicotinic acetylcholine receptor (nAChR) agonist, dose dependently attenuates Delta(9) tetrahydrocannabinol (Delta(9)THC)-induced ataxia. Presently, we have shown that intracerebellar nicotine (1.25, 2.5, and 5 ng; once daily for 5 days) and RJR 2403 (250, 500, and 750 ng; once daily for 5 days) significantly attenuate cerebellar Delta(9)-THC-induced ataxia dose dependently, suggesting the development of cross-tolerance between nicotine or RJR-2403 with Delta(9)-THC in male CD-1 mice. Intracerebellar RJR-2403 (750 ng) microinfused for 1, 2, 3, 5, and 7 days (once daily) significantly attenuated Delta(9)-THC-induced ataxia in the 3-, 5-, and 7-day treatment groups; optimal cross-tolerance was evident at day 5 and persisted till 36 h after the last RJR-2403 microinfusion. Intracerebellar microinfusion of hexamethonium (nAChR antagonist; 1 microg) or dihydro-beta-erythroidine hydrobromide (alpha(4)beta(2) nAChR antagonist; 500 ng) for 5 days 10 min before daily intracerebellar nicotine or RJR-2403 microinfusion virtually abolished cross-tolerance between nicotine or RJR-2403 and Delta(9) THC, indicating nAChR participation. In addition, microinfusion of antagonists 10 min after daily intracerebellar nicotine or RJR-2403 failed to alter the cross tolerance, suggesting possible involvement of downstream cerebellar second messenger mechanisms. Finally, the cerebellar concentration of nitric oxide products [total sum of nitrite + nitrate (NO(x))] was increased after 5 days of intracerebellar nicotine or RJR-2403 treatment, which was decreased by acute intracerebellar Delta(9)-THC treatment. The "nicotine or RJR-2403 + Delta(9)-THC" treatments significantly increased cerebellar NO(x) levels compared with treatment with Delta(9)-THC alone, supporting a functional correlation between cerebellar nitric oxide production and cerebellar Delta(9)-THC-induced ataxia and suggesting participation of nitric oxide in the observed cross-tolerance between nicotine/RJR-2403 and Delta(9)-THC. PMID- 17416742 TI - A novel family of potent negative allosteric modulators of group II metabotropic glutamate receptors. AB - Group II metabotropic glutamate receptors (mGluRs), mGluR2 and mGluR3, play a number of important roles in mammalian brain and represent exciting new targets for certain central nervous system disorders. We now report synthesis and characterization of a novel family of derivatives of dihydrobenzo[1,4]diazepin-2 one that are selective negative allosteric modulators for group II mGluRs. These compounds inhibit both mGluR2 and mGluR3 but have no activity at group I and III mGluRs. The novel mGluR2/3 antagonists also potently block mGluR2/3-mediated inhibition of the field excitatory postsynaptic potentials at the perforant path synapse in hippocampal slices. These compounds induce a rightward shift and decrease the maximal response in the glutamate concentration-response relationship, consistent with a noncompetitive antagonist mechanism of action. Furthermore, radioligand binding studies revealed no effect on binding of the orthosteric antagonist [(3)H]LY341495 [2S-2-amino-2-(1S,2S-2-carboxycyclopropan-1 yl)-3-(xanth-9-yl)propionic acid]. Site-directed mutagenesis revealed that a single point mutation in transmembrane V (N735D), previously shown to be an important residue for potentiation activity of the mGluR2 allosteric potentiator LY487379 [N-(4-(2-methoxyphenoxy)phenyl)-N-(2,2,2-trifluoroethylsulfonyl)pyrid-3 ylmethylamine], is not critical for the inhibitory activity of negative allosteric modulators of group II mGluRs. However, this single mutation in human GluR2 almost completely blocked the enhancing activity of biphenyl-indanone A, a novel allosteric potentiator of mGluR2. Our data suggest that these two positive allosteric modulators of mGluR2 may share a common binding site and that this site may be distinct from the binding site for the new negative allosteric modulators of group II mGluRs. PMID- 17416743 TI - PolyScan: an automatic indel and SNP detection approach to the analysis of human resequencing data. AB - Small insertions and deletions (indels) and single nucleotide polymorphisms (SNPs) are common genetic variants that are thought to be associated with a wide variety of human diseases. Owing to the genome's size and complexity, manually characterizing each one of these variations in an individual is not practical. While significant progress has been made in automated single-base mutation discovery from the sequences of diploid PCR products, automated and reliable detection of indels continues to pose difficult challenges. In this paper, we present PolyScan, an algorithm and software implementation designed to provide de novo heterozygous indel detection and improved SNP identification in the context of high-throughput medical resequencing. Tests on a human diploid PCR-based sequence data set, consisting of 90,270 traces from 13 genes, indicate that PolyScan identified approximately 90% of the 151 consensus indel sites and approximately 84% of the 1546 heterozygous indels previously identified by manual inspection. Tests on tumor-derived data show that PolyScan better identifies high quality, low-level mutations as compared with other mutation detection software. Moreover, SNP identification improves when reprocessing the results of other programs. These results suggest that PolyScan may play a useful role in the post human genome project research era. PMID- 17416744 TI - Rapid evolution of an X-linked microRNA cluster in primates. AB - MicroRNAs (miRNAs) are a growing class of small RNAs (about 22 nt) that play crucial regulatory roles in the genome by targeting mRNAs for cleavage or translational repression. Most of the identified miRNAs are highly conserved among species, indicating strong functional constraint on miRNA evolution. However, nonconserved miRNAs may contribute to functional novelties during evolution. Recently, an X-linked miRNA cluster was reported with multiple copies in primates but not in rodents or dog. Here we sequenced and compared this miRNA cluster in major primate lineages including human, great ape, lesser ape, Old World monkey, and New World monkey. Our data indicate rapid evolution of this cluster in primates including frequent tandem duplications and nucleotide substitutions. In addition, lineage-specific substitutions were observed in human and chimpanzee, leading to the emergence of potential novel mature miRNAs. The expression analysis in rhesus monkeys revealed a strong correlation between miRNA expression changes and male sexual maturation, suggesting regulatory roles of this miRNA cluster in testis development and spermatogenesis. We propose that, like protein-coding genes, miRNA genes involved in male reproduction are subject to rapid adaptive changes that may contribute to functional novelties during evolution. PMID- 17416745 TI - Fine mapping of a swine quantitative trait locus for number of vertebrae and analysis of an orphan nuclear receptor, germ cell nuclear factor (NR6A1). AB - The number of vertebrae in pigs varies and is associated with meat productivity. Wild boars, which are ancestors of domestic pigs, have 19 vertebrae. In comparison, European commercial breeds have 21-23 vertebrae, probably owing to selective breeding for enlargement of body size. We previously identified two quantitative trait loci (QTL) for the number of vertebrae on Sus scrofa chromosomes (SSC) 1 and 7. These QTL explained an increase of more than two vertebrae. Here, we performed a map-based study to define the QTL region on SSC1. By using three F2 experimental families, we performed interval mapping and recombination analyses and defined the QTL within a 1.9-cM interval. Then we analyzed the linkage disequilibrium of microsatellite markers in this interval and found that 10 adjacent markers in a 300-kb region were almost fixed in European commercial breeds. Genetic variation of the markers was observed in Asian local breeds or wild boars. This region encoded an orphan nuclear receptor, germ cell nuclear factor (NR6A1, formerly known as GCNF), which contained an amino acid substitution (Pro192Leu) coincident with the QTL. This substitution altered the binding activity of NR6A1 to its corepressors, nuclear receptor associated protein 80 (RAP80) and nuclear receptor corepressor 1 (NCOR1). In addition, somites of mouse embryos demonstrated expression of NR6A1 protein. Together, these results suggest that NR6A1 is a strong candidate for one of the QTL that influence number of vertebrae in pigs. PMID- 17416746 TI - Efficient target-selected mutagenesis in Caenorhabditis elegans: toward a knockout for every gene. AB - Reverse genetic or gene-driven knockout approaches have contributed significantly to the success of model organisms for fundamental and biomedical research. Although various technologies are available for C. elegans, none of them scale very well for genome-wide application. To address this, we implemented a target selected knockout approach that is based on random chemical mutagenesis and detection of single nucleotide mutations in genes of interest using high throughput resequencing. A clonal library of 6144 EMS-mutagenized worms was established and screened, resulting in the identification of 1044 induced mutations in 109 Mbp, which translates into an average spacing between exonic mutations in the library of only 17 bp. We covered 25% of the open reading frames of 32 genes and identified one or more inactivating mutations (nonsense or splice site) in 84% of them. Extrapolation of our results indicates that nonsense mutations for >90% of all C. elegans genes are present in the library. To identify all of these mutations, one only needs to inspect those positions that- given the known specificity of the mutagen--can result in the introduction of a stop codon. We define these positions as nonsense introducing mutations (NIMs). The genome-wide collection of possible NIMs can be calculated for any organism with a sequenced genome and reduces the screening complexity by 200- to 2000 fold, depending on the organism and mutagen. For EMS-mutagenized C. elegans, there are only approximately 500,000 NIMs. We show that a NIM genotyping approach employing high-density microarrays can, in principle, be used for the genome-wide identification of C. elegans knockouts. PMID- 17416747 TI - Fast-X on the Z: rapid evolution of sex-linked genes in birds. AB - Theoretical work predicts natural selection to be more efficient in the fixation of beneficial mutations in X-linked genes than in autosomal genes. This "fast-X effect" should be evident by an increased ratio of nonsynonymous to synonymous substitutions (dN/dS) for sex-linked genes; however, recent studies have produced mixed support for this expectation. To make an independent test of the idea of fast-X evolution, we focused on birds, which have female heterogamety (males ZZ, females ZW), where analogous arguments would predict a fast-Z effect. We aligned 2.8 Mb of orthologous protein-coding sequence of zebra finch and chicken from 172 Z-linked and 4848 autosomal genes. Zebra finch data were in the form of EST sequences from brain cDNA libraries, while chicken genes were from the draft genome sequence. The dN/dS ratio was significantly higher for Z-linked (0.110) than for all autosomal genes (0.085; P=0.002), as well as for genes linked to similarly sized autosomes 1-10 (0.0948; P=0.04). This pattern of fast-Z was evident even after we accounted for the nonrandom distribution of male-biased genes. We also examined the nature of standing variation in the chicken protein coding regions. The ratio of nonsynonymous to synonymous polymorphism (pN/pS) did not differ significantly between genes on the Z chromosome (0.104) and on the autosomes (0.0908). In conjunction, these results suggest that evolution proceeds more quickly on the Z chromosome, where hemizygous exposure of beneficial nondominant mutations increases the rate of fixation. PMID- 17416748 TI - Identification of mouse Duffy antigen receptor for chemokines (Darc) as a BMD QTL gene. AB - It is now well known that bone mineral density (BMD) variance is determined by both genetic and environmental factors. Accordingly, studies in human and animal models have revealed evidence for the presence of several quantitative trait loci (QTL) that contribute to BMD variations. However, the identification of BMD QTL genes remains a big challenge. In the current study, we focused our efforts to identify the BMD candidate gene in chromosome 1 (Chr 1) QTL that was detected from a cross involving high BMD CAST/EiJ (CAST) and low BMD C57BL/6J (B6) mice. To this end, we have combined several approaches including: (1) fine mapping the BMD QTL in Chr 1 of the B6.CAST F2 female mice using a large number of polymorphic markers; (2) the generation of congenic sublines of mice by repeated backcrossing of CAST with B6 mice and phenotype characterization; (3) expression profiling genes in the QTL region; and (4) SNP analyses to identify the mouse Duffy Antigen Receptor for Chemokines (Darc) as a candidate gene for Chr 1 BMD QTL2. We verified the involvement of the Darc protein in BMD variation by evaluating the skeletal phenotype of Darc-knockout mice and congenic sublines of mice carrying small chromosomal segments from CAST BMD QTL. Based on the findings that Darc-antibody blocked formation of multinucleated osteoclasts in vitro and that Darc from CAST binds chemokines, known to regulate osteoclast formation, with reduced affinity compared with Darc from B6 mice, we conclude that Darc regulates BMD negatively by increasing osteoclast formation, and that the genetic association between Darc gene polymorphism and BMD variations in humans merits investigation. PMID- 17416749 TI - Distinct mechanisms for trans-mediated mobilization of cellular RNAs by the LINE 1 reverse transcriptase. AB - Long Interspersed Element-1 (LINE-1 or L1) sequences comprise approximately 17% of human DNA and ongoing L1 retrotransposition continues to impact genome evolution. The L1-encoded proteins also can mobilize other cellular RNAs (e.g., Alu retrotransposons, SVA retrotransposons, and U6 snRNAs), which comprise approximately 13% of human DNA. Here, we demonstrate that the trans-mediated mobilization of non-L1 RNAs can occur by either template choice or template switching mechanisms. Remarkably, these mechanisms are not mutually exclusive, as both processes can operate sequentially on the same RNA template. Finally, we provide evidence that efficient U6 snRNA retrotransposition requires both ORF1p and ORF2p, providing indirect evidence for the action of ORF1p in U6 snRNA retrotransposition. Thus, we propose that the LINE-1-encoded reverse transcriptase can mediate the retrotransposition of non-L1 RNAs by distinct mechanisms. PMID- 17416751 TI - Microarrays and epidemiology: not the beginning of the end but the end of the beginning... PMID- 17416750 TI - X chromosome cDNA microarray screening identifies a functional PLP2 promoter polymorphism enriched in patients with X-linked mental retardation. AB - X-linked Mental Retardation (XLMR) occurs in 1 in 600 males and is highly genetically heterogeneous. We used a novel human X chromosome cDNA microarray (XCA) to survey the expression profile of X-linked genes in lymphoblasts of XLMR males. Genes with altered expression verified by Northern blot and/or quantitative PCR were considered candidates. To validate this approach, we documented the expected changes of expression in samples from a patient with a known X chromosome microdeletion and from patients with multiple copies of the X chromosome. We used our XCA to survey lymphoblast RNA samples from 43 unrelated XLMR males and found 15 genes with significant (>or=1.5-fold) reduction in expression in at least one proband. Of these, subsequent analysis confirmed altered expression in 12. We followed up one, PLP2, at Xp11.23, which exhibits approximately fourfold decreased expression in two patients. Sequencing analysis in both patients revealed a promoter variant, -113C>A, that alters the core binding site of the transcription factor ELK1. We showed that PLP2-(-113C>A) is sufficient to cause reduced expression using a luciferase reporter system and is enriched in a cohort of males with probable XLMR (14 of 239, 5.85%) as compared to normal males (9 of 577, 1.56%) (chi2=11.07, P<0.001). PLP2 is expressed abundantly in the pyramidal cells of hippocampus and granular cells of the cerebellum in the brain. We conclude that our XCA screening is an efficient strategy to identify genes that show significant changes in transcript abundance as candidate genes for XLMR. PMID- 17416752 TI - Physical activity and endometrial cancer risk, a systematic review of current evidence. AB - OBJECTIVE: To assess the epidemiologic evidence for the association between physical activity and endometrial cancer risk, taking into account the methodologic quality of each study. DESIGN: Systematic review, best evidence synthesis. DATA SOURCES: Studies were identified through a systematic review of literature available on PubMed through December 2006. REVIEW METHODS: We included cohort and case-control studies that assessed total and/or leisure time and/or occupational activities in relation to the incidence of endometrial cancer. The methodologic quality of the studies was assessed with a comprehensive scoring system. RESULTS: The included cohort (n = 7) and case-control (n = 13) studies consistently show that physical activity is associated with a decreased risk of endometrial cancer. The best evidence synthesis showed that the majority (80%) of 10 high-quality studies found risk reductions of >20%. Pooling of seven high quality cohort studies that measured total, leisure time, or occupational activity showed a significantly decreased risk of endometrial cancer (summary estimate: OR, 0.77; 95% CI, 0.70-0.85) for the most active women. Case control studies with relatively unfavorable quality scores reported divergent risk estimates, between 2-fold decreased and 2-fold increased risk. Effect modification by body mass index or menopausal status was not consistently observed. Evidence for an effect of physical activity during childhood or adolescence was limited. CONCLUSIONS: Physical activity seems to be associated with a reduction in the risk of endometrial cancer, which is independent of body weight. Further studies, preferably prospective cohort studies, are needed to determine the magnitude of the risk reduction and to assess which aspects of physical activity contribute most strongly to the reduced risk and in which period of life physical activity is most effective. PMID- 17416753 TI - S100A6 is increased in a stepwise manner during pancreatic carcinogenesis: clinical value of expression analysis in 98 pancreatic juice samples. AB - There are few reports describing the diagnostic significance of S100A6 expression in clinical samples obtained from patients with pancreatic disease. In the present study, we measured S100A6 expression in pancreatic tissues and juice to evaluate its involvement in pancreatic carcinogenesis. We did quantitative real time reverse transcription-PCR to measure mRNA expression in microdissected cells and pancreatic juice samples. Microdissected invasive ductal carcinoma and intraductal papillary mucinous neoplasm (IPMN) cells expressed significantly higher levels of S100A6 than did microdissected pancreatitis-affected epithelial and normal cells (all comparison; P < 0.008). Median levels of S100A6 in invasive ductal carcinoma were higher than those in IPMN, and those in pancreatitis affected epithelial cells tended to be higher than those in normal cells, although these differences were not statistically significant. In analyses of pancreatic juice, IPMN and pancreatic cancer samples expressed significantly higher levels of S100A6 than did chronic pancreatitis samples (both; P < 0.017), but levels in pancreatic cancer and IPMN samples did not differ form each other. Receiver operating characteristic (ROC) curve analysis revealed that measurement of S100A6 was useful for discriminating cancer (area under the ROC curve, 0.864) or IPMN (area under the ROC curve, 0.749) from chronic pancreatitis. The present data suggest that expression of S100A6 is increased in a stepwise manner during pancreatic carcinogenesis and may be a biomarker for evaluating malignant potential. Measurement of S100A6 in pancreatic juice may be useful to detect early pancreatic cancer or identify individuals with high-risk lesions that may progress to pancreatic cancer. PMID- 17416754 TI - Dietary mutagen exposure and risk of pancreatic cancer. AB - To investigate the association between dietary exposure to food mutagens and risk of pancreatic cancer, we conducted a hospital-based case-control study at the University of Texas M. D. Anderson Cancer Center during June 2002 to May 2006. A total of 626 cases and 530 noncancer controls were frequency matched for race, sex and age (+/-5 years). Dietary exposure information was collected via personal interview using a meat preparation questionnaire. A significantly greater portion of the cases than controls showed a preference to well-done pork, bacon, grilled chicken, and pan-fried chicken, but not to hamburger and steak. Cases had a higher daily intake of food mutagens and mutagenicity activity (revertants per gram of daily meat intake) than controls did. The daily intakes of 2-amino-3,4,8 trimethylimidazo[4,5-f]quinoxaline (DiMeIQx) and benzo(a)pyrene (BaP), as well as the mutagenic activity, were significant predictors for pancreatic cancer (P = 0.008, 0.031, and 0.029, respectively) with adjustment of other confounders. A significant trend of elevated cancer risk with increasing DiMeIQx intake was observed in quintile analysis (P(trend) = 0.024). A higher intake of dietary mutagens (those in the two top quintiles) was associated with a 2-fold increased risk of pancreatic cancer among those without a family history of cancer but not among those with a family history of cancer. A possible synergistic effect of dietary mutagen exposure and smoking was observed among individuals with the highest level of exposure (top 10%) to PhIP and BaP, P(interaction) = 0.09 and 0.099, respectively. These data support the hypothesis that dietary mutagen exposure alone and in interaction with other factors contribute to the development of pancreatic cancer. PMID- 17416755 TI - Gastric cancer is related to early Helicobacter pylori infection in a high prevalence country. AB - BACKGROUND AND AIMS: Chile ranks fifth in the world among countries with the highest incidence of gastric cancer. The aim was to quantify the association between Helicobacter pylori infection and gastric cancer mortality at the county of residence. METHODS: A cross-sectional household survey, a probability sample of the Chilean adult population, provided 2,615 participants in whom serum H. pylori IgG antibodies were measured (ELISA). The spatial pattern of 48,367 deaths due to gastric cancer which occurred from 1985 to 2002 was analyzed using a hierarchical Poisson regression model; 333 counties were categorized as low, medium, and high gastric cancer mortality with median gastric cancer death rates of 11.4, 19.1, and 26.0 per 100,000 inhabitants, respectively. The association between H. pylori positivity and gastric cancer mortality in the county of residence was assessed by multivariate Poisson regression for complex samples. RESULTS: H. pylori prevalence was 73.0% [95% confidence intervals (CI), 70.0 76.0], higher in men [prevalence rate ratio (PRR), 1.1 (95% CI, 1.01-1.20)], peaked at ages 45 to 64, and dropped after age 65. It was higher among residents in counties with high gastric cancer mortality (79.7%; 95% CI, 76.4-82.6) compared to counties with low gastric cancer mortality (62.3%; 95% CI, 53.8-70.2; corresponding PRR, 1.3; 95% CI, 1.1-1.5); under age 24, H. pylori infection was 79.7% (95% CI, 72.2-85.6) versus 39.8% (95% CI, 19.6-64.2) among residents in counties with high and low gastric cancer mortalities, respectively (PRR, 2.0; 95% CI, 1.1-3.7). CONCLUSIONS: The high prevalence of H. pylori at younger ages was associated with high gastric cancer mortality in the base population. PMID- 17416756 TI - Dietary fiber and risk of colorectal cancer in the Japan collaborative cohort study. AB - To examine the association of dietary fiber with the risk of colorectal cancer in a population with a high incidence of cancer and a low fiber intake, we analyzed the data from the Japan Collaborative Cohort Study. From 1988 to 1990, 43,115 men and women aged 40 to 79 years completed a questionnaire on dietary and other factors. Intake of dietary fiber was estimated using a food frequency questionnaire. Rate ratios (RR) were computed by fitting proportional hazards models. During the mean follow-up of 7.6 years, 443 colorectal cancer cases were recorded. In all participants, we found a decreasing trend in risk of colorectal cancer with increasing intake of total dietary fiber; the multivariate-adjusted RRs across quartiles were 1.00, 0.96 [95% confidence interval (95% CI), 0.72 1.27], 0.72 (0.53-0.99), and 0.73 (0.51-1.03; P(trend) = 0.028). This trend was exclusively detected for colon cancer: the corresponding RRs were 1.00, 0.90 (95% CI, 0.64-1.26), 0.56 (0.38-0.83), and 0.58 (0.38-0.88; P(trend) = 0.002). The decrease in RRs with increasing intake of dietary fiber was larger in men than in women. No material differences appeared in the strength of associations with the risk between water-soluble and insoluble dietary fiber. For food sources of fiber, bean fiber intake was somewhat inversely correlated with colorectal cancer risk. This prospective study supported potential protective effects of dietary fiber against colorectal cancer, mainly against colon cancer. The role of dietary fiber in the prevention of colorectal cancer seems to remain inconsistent, and further investigations in various populations are warranted. ( PMID- 17416757 TI - Do factors related to endogenous and exogenous estrogens modify the relationship between obesity and risk of colorectal adenomas in women? AB - Obesity has consistently been associated with increased colorectal cancer risk in men, but not in women. In the absence of postmenopausal hormone use (PMH), adipose-derived estrogen is the primary determinant of circulating estrogen levels in postmenopausal women, perhaps ameliorating the mitogenic effects of obesity in this group. Using data from a case-control study in the United States, we examined associations among obesity, potential modifying effects of factors related to endogenous and exogenous estrogen levels, and risk of colorectal adenoma. Cases (n = 219) were women of ages 30 to 74 years with colonoscopy proven, incident, sporadic, pathology-confirmed, adenomatous polyps of the colon and rectum. Two control groups were recruited: colonoscopy-confirmed polyp-free women (n = 438) and age- and zip code frequency-matched women randomly selected from the community (n = 247). Multivariate odds ratios and 95% confidence intervals (95% CI) for obese [body mass index (BMI) >or=30.0; compared with nonobese, BMI <25.0] premenopausal women were 2.09 (95% CI, 0.81-5.41) versus colonoscopy controls, and 5.18 (95% CI, 1.40-19.32) versus population controls. For PMH users, the corresponding odds ratios were 0.29 (95% CI, 0.12-0.70) versus colonoscopy controls and 0.64 (95% CI, 0.23-1.83) versus population controls. There was no significant association of BMI with adenoma risk for PMH nonusers. Findings for waist-to-hip ratio were similar to those for BMI. These data support the hypothesis that risk for colorectal adenoma may be increased with obesity among premenopausal women but decreased among postmenopausal women, especially if they also take PMH. PMID- 17416758 TI - Dietary flavonoids and the risk of colorectal cancer. AB - In vitro and in vivo laboratory data point to chemoprotective effects of flavonoids on colorectal cancer. However, there has been limited epidemiologic research on the dietary intake of flavonoids and risk of colorectal cancer. Recent expansions of dietary databases to include flavonoid data now make such studies feasible. Association between the six main classes of flavonoids and the risk of colorectal cancer was examined using data from a national prospective case-control study in Scotland, including 1,456 incident cases and 1,456 population-based controls matched on age, sex, and residence area. Dietary, including flavonoid data, were obtained from a validated, self-administered food frequency questionnaire. Risk of colorectal cancer was estimated using conditional logistic regression models in the whole sample and stratified by sex, smoking status, and cancer site and adjusted for established and putative risk factors. After energy adjustment, reductions in colorectal cancer risk associated with the highest quartiles of intake (versus the lowest quartile) were 27% for flavonols [odds ratio (OR), 0.73; P(trend) = 0.012], 32% for quercetin (OR, 0.68; P(trend) = 0.001), 32% for catechin (OR, 0.68; P(trend) < 0.0005); 26% for epicatechin (OR, 0.74; P(trend) = 0.019), and 22% for procyanidins (OR, 0.78; P(trend) = 0.031). The significant dose-dependent reductions in colorectal cancer risk that were associated with increased consumption of flavonols, quercetin, catechin, and epicatechin remained robust after controlling for overall fruit and vegetable consumption or for other flavonoid intake. The risk reductions were greater among nonsmokers, but no interaction beyond a multiplicative effect was present. Sex-specific or cancer-type differences were not observed. No risk reductions were associated with intake of flavones (P(trend) = 0.64), flavonones (P(trend) = 0.22), and phytoestrogens (P(trend) = 0.26). This was the first of several a priori hypotheses to be tested in this large study and showed strong and linear inverse associations of flavonoids with colorectal cancer risk. PMID- 17416759 TI - Perceived risk of colorectal cancer: sources of risk judgments. AB - Perceived risk of disease plays a key role in health behaviors, making it an important issue for cancer-preventive behavior research. This study describes studies using complimentary methodologies to investigate the determinants of perceived risk of developing colorectal cancer. In study 1, questionnaires were mailed to a community sample of 1,056 adults ages 45 to 65 years. They assessed risk factors for colorectal cancer and perceived risk of colorectal cancer and asked respondents to explain why they had rated their risk as they had. Consistent with previous studies, risk factors for colorectal cancer were significantly correlated with perceived risk, although associations were relatively weak. The most frequently cited reasons for risk judgments were diet, family history, and symptoms/general health. Not surprisingly, correlations between each risk factor and perceived risk were stronger among participants who had attributed their risk to that factor. Study 2 used semi-structured interviews to explore individuals' explanations for their perceived risk of colorectal cancer. Adults (n = 18) ages 60 to 63 years volunteered a variety of explanations, but their experiences of cancer seemed to be most salient to their risk estimates. These studies suggest that attributions people make for their risk judgments are important in understanding how they conceptualize risk, but appreciation of experiential and affective influences may be useful in fully understanding perceptions of risk. PMID- 17416760 TI - Insulin resistance-related genes and advanced left-sided colorectal adenoma. AB - BACKGROUND: Insulin resistance has been linked with colorectal neoplasia through a number of mechanistic and observational studies. Allelic variants of genes encoding components of the insulin pathway, including insulin (INS), insulin receptor (INSR), and insulin receptor substrate-1 and insulin receptor substrate 2 (IRS1 and IRS2) have been associated with hyperinsulinemia and insulin resistance and may, therefore, predict susceptibility to colorectal neoplasia. METHODS: We investigated whether single nucleotide polymorphisms (SNP) in the INS, INSR, IRS1, and IRS2 genes are associated with risk of advanced left-sided colorectal adenoma, a cancer precursor. We analyzed 20 SNPs in a largely Caucasian study population comprising 766 cases with advanced adenomas of the distal colon and 771 controls, all of whom had undergone flexible sigmoidoscopy as part of the screening arm of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. RESULTS: Overall, we found limited evidence for a role of gene variants of the insulin signaling pathway and prevalence of advanced colorectal adenoma. We observed a statistically significant interaction between INSR genotypes and body mass index (BMI) with colorectal adenoma prevalence (P value for global test = 0.003) and suggestion of an interaction between INSR genotypes and glycemic load (P value for global test = 0.06); however, exploration of the interaction of BMI and glycemic load with the individual SNPs in INSR did not suggest a single SNP that may explain the significance of these global tests of interaction and did not yield any consistent patterns. CONCLUSION: These findings do not provide strong evidence for associations between polymorphic variation in genes of the insulin signaling pathway and advanced left-sided colorectal adenoma. Evidence for interaction between INSR variants and BMI and glycemic load for risk of advanced left-sided colorectal adenoma requires independent confirmation, and genotyping of INSR across a broader region and at greater density may be necessary to fully elucidate the nature of these interactions. PMID- 17416761 TI - Incidence and duration of cervical human papillomavirus 6, 11, 16, and 18 infections in young women: an evaluation from multiple analytic perspectives. AB - OBJECTIVE: To estimate the incidence and duration of cervical human papillomavirus (HPV)-6, HPV-11, HPV-16, and HPV-18 infections in a population of young American women. METHODS: The study population consisted of U.S. women who at baseline were 16 to 23 years of age, reported zero to five lifetime sexual partners, never having been pregnant, and never having had a prior abnormal Papanicolaou test and were enrolled in the placebo arm of a randomized multicenter clinical trial of a HPV-16 L1 virus-like particle vaccine. Women underwent type-specific endocervical/ectocervical swab HPV DNA testing at approximately 6-month intervals for up to 48 months of follow-up. To contribute person-time in the analyses of type-specific HPV incidence, a woman must have had at least three satisfactory swab specimens available and been negative for the relevant HPV type (HPV-6, HPV-11, HPV-16, or HPV-18) on her first two trial swabs. The duration of incident HPV infections was estimated using Kaplan-Meier survival analysis methods. RESULTS: Person-years of exposure ranged by type specific analysis from 2,645 to 3,188, with an incidence rate per 100 person years of 3.6 for HPV-6, 0.4 for HPV-11, 5.4 for HPV-16, and 2.1 for HPV-18. With censoring at the time of treatment for cervical intraepithelial neoplasia, where done, the mean duration of incident infections was 9.3, 8.4, 18.2, and 16.4 months, respectively, for HPV-6 (n = 103), HPV-11 (n = 13), HPV-16 (n = 142), and HPV-18 (n = 62). When the duration of HPV infections was truncated at the time of cervical intraepithelial neoplasia detection (any grade), where applicable, mean duration figures were 8.4, 8.1, 14.0, and 15.1 months for HPV-6, HPV-11, HPV-16, and HPV-18 infections, respectively. CONCLUSIONS: Previous studies of the mean duration of cervical HPV infection have been based on prevalent infections and/or featured relatively short duration of follow-up. This study tested women for HPV infection over a period of up to 48 months and observed a mean duration of incident HPV-16/HPV-18 infections approximately twice that of HPV-6/HPV-11. PMID- 17416762 TI - Insulin-like growth factor-I and risk of high-grade cervical intraepithelial neoplasia. AB - Insulin-like growth factors (IGF) and their binding proteins (IGFBP) have been implicated in the risk of several epithelial or glandular tumors, including prostate cancer, breast cancer, and colon cancer. Cervical cancer, which is also of epithelial origin, has been shown to overexpress receptors for IGF-I, and plasma levels of IGF-I have been positively associated with cervical cancer precursors in one epidemiologic study. In this case-control study, we investigated plasma levels of IGF-I and IGFBP-3 in relation to the risk of histologically confirmed high-grade cervical intraepithelial neoplasia (HGCIN) and the risk of human papillomavirus (HPV) infection. Included in this analysis were 329 cases and 621 controls recruited from clinics affiliated with two Montreal-area hospital centers. We observed a reduced risk of HGCIN for increasing levels of IGF-I, with an adjusted odds ratio (OR) of 0.40 (95% confidence interval, 0.19-0.87) for the highest quartile relative to the lowest quartile of IGF-I. No association was observed between IGFBP-3 levels and HGCIN. Among controls, IGF-I was associated with a decreased risk of being positive for HPV-16 or HPV-18, with an adjusted odds ratio of 0.20 (95% confidence interval, 0.05-0.87) for the highest quartile relative to the lowest quartile of IGF-I. There was no association observed between IGFBP-3 levels and HPV infection status. IGF-I-mediated effects seemed to predominate among women <30 years of age. In contrast to the previously reported study, our results suggest that levels of IGF-I in young women may be inversely associated with HGCIN, a precursor to cervical cancer. PMID- 17416763 TI - Lifetime weight history and endometrial cancer risk by type of menopausal hormone use in the NIH-AARP diet and health study. AB - Obesity and menopausal estrogen therapy are established risk factors for endometrial cancer. However, the joint effects of obesity and menopausal hormone therapy on endometrial cancer risk are incompletely understood. We addressed this issue in a cohort of 103,882 women ages 50 to 71 years at baseline in 1995 to 1996. During a median of 4.6 years, which contributed to a total of 455,304 person-years of follow-up through 2000, 677 cases of endometrial cancer were ascertained. Both baseline body mass index (BMI) and adult weight gain were associated with increased endometrial cancer risk. The multivariate relative risk (RR) comparing obese with normal weight women (BMI >30 versus <25 kg/m(2)) was 3.03 [95% confidence interval (95% CI), 2.50-3.68]. Compared with women with stable weight (gained or lost <5 kg) between age 18 and baseline, women who gained >or=20 kg had a RR of 2.75 (95% CI, 1.96-3.86). Menopausal hormone therapy significantly modified the relations of BMI (P(interaction) < 0.001) and adult weight gain (P(interaction) = 0.004) to endometrial cancer risk. Compared with normal weight, the RRs for obesity were 5.41 (95% CI, 4.01-7.29) among women who never used menopausal hormone therapy, 2.53 (95% CI, 1.21-5.30) among former menopausal hormone therapy users, and 1.44 (95% CI, 1.00-2.05) among current users. Compared with a stable weight between age 18 and baseline, the RRs for weight gain of >or=20 kg among never users and ever users of menopausal hormone therapy were 5.35 (95% CI, 3.01-9.52) and 1.43 (95% CI, 0.96-2.15), respectively. We conclude that both current adiposity and adult weight gain are associated with substantial increases in the risk of endometrial cancer, with relations particularly evident among never users of menopausal hormone therapy. PMID- 17416764 TI - Effects of aerobic exercise training on estrogen metabolism in premenopausal women: a randomized controlled trial. AB - BACKGROUND: Regular physical activity may alter estrogen metabolism, a proposed biomarker of breast cancer risk, by shifting metabolism to favor production of 2 hydroxyestrone (2-OHE1). Few studies, however, have examined this question using a randomized controlled trial. PURPOSE: To examine the effects of 12 weeks of aerobic exercise training on 2-OHE1 and 16alpha-hydroxyestrone (16alpha-OHE1) in premenopausal women. METHODS: Participants were healthy, regularly menstruating, Caucasian women, 20 to 35 years, body mass index of 18 to 29.9, not using pharmacologic contraceptives, with average or below average fitness [maximal oxygen consumption (VO(2max)), <40 mL/kg/min]. Following a baseline menstrual cycle, participants (N = 32) were randomly assigned to a 12-week aerobic exercise training intervention (n = 17) or usual lifestyle (n = 15). Height, body mass, body composition by dual-energy X-ray absorptiometry, and VO(2max) were measured at baseline and following the intervention. Urine samples were collected in the luteal phase of four consecutive menstrual cycles. RESULTS: The exercise group increased VO(2max) by 14% and had significant, although modest, improvements in fat and lean body mass. No significant between-group differences were observed, however, for the changes in 2-OHE1 (P = 0.944), 16alpha-OHE1 (P = 0.411), or the ratio of 2-OHE1 to 16alpha-OHE1 (P = 0.317). At baseline, there was an inverse association between body fat and 2-OHE1 to 16alpha-OHE1 ratio (r = -0.40; P = 0.044); however, it was the change in lean body mass over the intervention that was positively associated with a change in 2-OHE1 to 16alpha-OHE1 ratio (r = 0.43; P = 0.015). CONCLUSIONS: A 12-week aerobic exercise training intervention significantly improved aerobic fitness and body composition but did not alter estrogen metabolism in premenopausal women. Interestingly, an increase in lean body mass was associated with a favorable change in 2-OHE1 to 16alpha-OHE1 ratio. PMID- 17416765 TI - Age at menarche and menopause and breast cancer risk in the International BRCA1/2 Carrier Cohort Study. AB - BACKGROUND: Early menarche and late menopause are important risk factors for breast cancer, but their effects on breast cancer risk in BRCA1 and BRCA2 carriers are unknown. METHODS: We assessed breast cancer risk in a large series of 1,187 BRCA1 and 414 BRCA2 carriers from the International BRCA1/2 Carrier Cohort Study. Rate ratios were estimated using a weighted Cox-regression approach. RESULTS: Breast cancer risk was not significantly related to age at menopause [hazard ratio [HR] for menopause below age 35 years, 0.60 [95% confidence interval (95% CI), 0.25-1.44]; 35 to 40 years, 1.15 [0.65-2.04]; 45 to 54 years, 1.02 [0.65-1.60]; >or=55 years, 1.12 [0.12-5.02], as compared with premenopausal women]. However, there was some suggestion of a reduction in risk after menopause in BRCA2 carriers. There was some evidence of a protective effect of oophorectomy (HR, 0.56; 95% CI, 0.29-1.09) and a significant trend of decreasing risk with increasing time since oophorectomy, but no apparent effect of natural menopause. There was no association between age at menarche and breast cancer risk, nor any apparent association with the estimated total duration of breast mitotic activity. CONCLUSIONS: These results are consistent with other observations suggesting a protective effect of oophorectomy, similar in relative effect to that in the general population. The absence of an effect of age at natural menopause is, however, not consistent with findings in the general population and may reflect the different natural history of the disease in carriers. PMID- 17416766 TI - IL6, aspirin, nonsteroidal anti-inflammatory drugs, and breast cancer risk in women living in the southwestern United States. AB - Interleukin-6 is a cytokine thought to be involved in inflammation, insulin, and estrogen-related pathways. We evaluate genetic variation in the IL6 gene with risk of breast cancer. We also evaluate breast cancer associations with aspirin and nonsteroidal anti-inflammatory drugs. A breast cancer case-control study (n = 1,527 non-Hispanic white cases, 1,601 non-Hispanic white controls, 798 Hispanic/Native American cases, and 924 Hispanic/Native American controls) was conducted among women living in the southwestern United States (4-Corner's Breast Cancer Study). Five IL6 single nucleotide polymorphisms (SNP) and IL6 haplotypes based on these SNPs were evaluated. Allele frequencies were significantly different between non-Hispanic white and Hispanic/Native American women. Among postmenopausal women not recently exposed to hormones, the AG/GG genotypes of rs1800797 (-596A>G) and the GC/CC genotypes of rs1800795 (-174G>C) significantly reduced risk of breast cancer among non-Hispanic white women [odds ratio (OR), 0.69; 95% confidence interval (95% CI), 0.48-1.00 and OR, 0.68; 95% CI, 0.47 0.99, respectively] and Hispanic/Native American women (OR, 0.48; 95% CI, 0.28 0.83 and OR, 0.44; 95% CI, 0.26-0.99, respectively). Haplotypes of the five IL6 SNPs further defined these associations. Recent aspirin use significantly decreased risk of breast cancer among postmenopausal Hispanic/Native American women not recently exposed to hormones (OR, 0.56; 95% CI, 0.33-0.96). Among non Hispanic white, the inverse association with aspirin was not statistically significant. IL6 genotype and haplotype significantly modified the association between aspirin and breast cancer, with the greatest effect modification being among women not recently exposed to hormones [P interaction = 0.06 (for non Hispanic white) and 0.04 (for Hispanic/Native American) and SNP rs1800796 or 572G>C]. These data suggest that IL6 is associated with breast cancer risk and modifies the association between estrogen and aspirin and breast cancer risk. PMID- 17416767 TI - Detecting an association between socioeconomic status and late stage breast cancer using spatial analysis and area-based measures. AB - OBJECTIVES: To assess the relationship between socioeconomic status (SES) and late stage breast cancer using the cluster detection software SaTScan and U.S. census-derived area-based socioeconomic measures. MATERIALS AND METHODS: Florida's 18,683 women diagnosed with late stage breast cancer (regional or distant stage) between 1998 and 2002 as identified by Florida's population-based, statewide, incidence registry were analyzed by SaTScan to identify areas of higher-than-expected incidence. The relationship between SES and late stage breast cancer was assessed at the neighborhood (block group) level by combining the SaTScan results with area-based SES data. RESULTS: SaTScan identified 767 of Florida's 9,112 block groups that had higher-than-expected incidence of late stage breast cancer. After controlling for patient level insurance status, county level mammography prevalence, and urban/rural residence in the logistic regression model, women living in neighborhoods of severe and near poverty were respectively 3.0 and 1.6 times more likely to live in areas of higher-than expected incidence of late stage breast cancer when compared with women living in nonpoverty. Additionally, areas in the lowest quartile of mammography usage were almost seven times more likely to have higher-than-expected incidence than areas in the higher quartiles. CONCLUSIONS: In addition to confirming the importance of mammography, results from the present study suggest that "where" you live plays an important role in defining the risk of presenting with late stage breast cancer. Additional research is urgently needed to understand this risk and to leverage the strengths and resources present in all communities to lower the late stage breast cancer burden. PMID- 17416768 TI - Circulating insulin-like growth factor-I and binding protein-3 and the risk of breast cancer. AB - Four meta-analyses and literature reviews have concluded that a positive association exists between circulating levels of insulin-like growth factor-I (IGF-I) and IGF-binding protein-3 (IGFBP-3) and breast cancer risk for premenopausal but not postmenopausal women. Recently, a large prospective study reported an association with IGF-I and IGFBP-3 concentration for breast cancer diagnosed after, but not before, the age of 50 years; and in a large cohort of primarily premenopausal women, IGF-I and IGFBP-3 were not associated with breast cancer risk. We did a case-cohort study within the Melbourne Collaborative Cohort Study, which included a random sample of 1,901 women (subcohort) and 423 breast cancer cases diagnosed during a mean of 9.1 years of follow-up. IGF-I and IGFBP-3 concentrations were measured in plasma collected at baseline. The association between quartiles of IGF concentration and breast cancer risk was tested using a Cox model adjusted for known and potential confounders. The hazard ratio (HR) for breast cancer comparing the fourth with the first quartiles was 1.20 [95% confidence interval (95% CI), 0.87-1.65] for IGF-I and 1.09 (95% CI, 0.78-1.53) for IGFBP-3. Both associations varied with age: for IGF-I, the HRs for breast cancer comparing the fourth with the first quartiles were 0.60 (95% CI, 0.25 1.45) before age 50 and 1.61 (95% CI, 1.04-2.51) after age 60 (test for the log linear trend of HR according to age, P = 0.05); for IGFBP-3, the HRs were 0.79 (95% CI, 0.34-1.83) before age 50 and 1.62 (95% CI, 1.03-2.55) after age 60 (test for log-linear trend, P = 0.08). IGF-I and IGFBP-3 were positively associated with breast cancer risk in older women but not in younger women. More prospective studies are needed to clarify the age dependence of the association between IGF-I and IGFBP-3 and breast cancer. PMID- 17416769 TI - A systematic approach to analysing gene-gene interactions: polymorphisms at the microsomal epoxide hydrolase EPHX and glutathione S-transferase GSTM1, GSTT1, and GSTP1 loci and breast cancer risk. AB - OBJECTIVE: We undertook a case-control study in an Australian Caucasian population-based sample of 1,246 cases and 664 controls to assess the roles of detoxification gene polymorphisms EPHX T>C Tyr(113)His, GSTT1 deletion, GSTM1 deletion, and GSTP1 A>G Ile(105)Val on risk of breast cancer. METHODS: We systematically addressed the main effects and possible gene-gene interactions using unconditional logistic regression to estimate odds ratios (OR) adjusted for potential confounders and using standard model building approaches based on likelihood theory. RESULTS: There was a decreased risk associated with the EPHX CC genotype [OR, 0.60; 95% confidence interval (95% CI), 0.43-0.84; P = 0.003], marginally significant evidence of increased risk with GSTM1 null genotype (OR, 1.21; 95% CI, 1.00-1.47; P = 0.05), but no association with GSTT1 null genotype (OR, 1.12; 95% CI, 0.86-1.45; P = 0.4) or GSTP1 (OR, 0.95; 95% CI, 0.82-1.10; P = 0.5) genotype. The full model with all interactions gave a significantly better fit than a main-effects-only model (P < 0.001), providing evidence for gene-gene interactions. The most parsimonious model included main effects for EPHX, GSTT1, and GSTM1; a two-way interaction between EPHX and GSTM1; and a three-way interaction between EPHX, GSTM1, and GSTT1. Predicted risks were greatest for women carrying deletions of both GSTT1 and GSTM1, with either the EPHX TC genotype (OR, 2.02; 95% CI, 1.19-3.45; P = 0.009) or EPHX CC genotype (OR, 3.54; 95% CI, 1.29-9.72; P = 0.14). CONCLUSION: Detoxification gene polymorphisms may interact with each other to result in small groups of individuals at modestly increased risk. We caution against overinterpretation and suggest that pooling of similarly large studies is needed to clarify the possible role of such complex gene-gene interactions on breast cancer risk. 2007;16(4):769-74). PMID- 17416770 TI - The epidemiology of arm and hand swelling in premenopausal breast cancer survivors. AB - BACKGROUND: Breast cancer survivors suffer from lymphedema of the arm and/or hand. Accurate estimates of the incidence and prevalence of lymphedema are lacking, as are the effects of this condition on overall quality of life. METHODS: Six hundred twenty-two breast cancer survivors (age, or=32 ng/mL) versus inadequate 25-hydroxyvitamin D levels had a modestly decreased risk of serous ovarian cancer (RR, 0.64; 95% CI, 0.39-1.05). Overall, our results do not suggest that plasma vitamin D levels are associated with risk of ovarian cancer. However, we observed significant associations in some subgroups, which should be evaluated further in other studies because increasing vitamin D intake is an easy preventive measure to adopt. PMID- 17416772 TI - Extracellular activity of cyclic AMP-dependent protein kinase as a biomarker for human cancer detection: distribution characteristics in a normal population and cancer patients. AB - The overexpression of cyclic AMP (cAMP)-dependent protein kinase (PKA) has been reported in patients with cancer, and PKA inhibitors have been tested in clinical trials as a novel cancer therapy. The present study was designed to characterize the population distribution of extracellular activity of cAMP-dependent protein kinase (ECPKA) and its potential value as a biomarker for cancer detection and monitoring of cancer therapy. The population distribution of ECPKA activity was determined in serum samples from a Chinese population consisting of a total of 603 subjects (374 normal healthy volunteers and 229 cancer patients). The serum ECPKA was determined by a validated sensitive radioassay, and its diagnostic values (including positive and negative predictive values) were analyzed. The majority of normal subjects (>70%) have undetectable or very low levels of serum ECPKA. In contrast, the majority of cancer patients (>85%) have high levels of ECPKA. The mean ECPKA activity in the sera of cancer patients was 10.98 units/mL, 5-fold higher than that of the healthy controls (2.15 units/mL; P < 0.001). In both normal subjects and cancer patients, gender and age had no significant influence on the serum ECPKA. Among factors considered, logistic analysis revealed that the disease (cancer) is the only factor contributing to the elevation of ECPKA activity in cancer patients. In conclusion, ECPKA may function as a cancer marker for various human cancers and can be used in cancer detection and for monitoring response to therapy with other screening or diagnostic techniques. PMID- 17416773 TI - Exposure to ethylene oxide in hospitals: biological monitoring and influence of glutathione S-transferase and epoxide hydrolase polymorphisms. AB - Ethylene oxide is considered as a human carcinogen. A biomarker of exposure would be a useful instrument to assess the risk in occupationally exposed workers. This cross-sectional study aimed at examining (a) whether the urinary excretion of a metabolite of ethylene oxide, 2-hydroxyethyl mercapturic acid (HEMA), could be used for monitoring occupational exposure and (b) whether glutathione S transferase (GST) and epoxide hydrolase genotypes influenced biological monitoring. Exposure to ethylene oxide was measured by personal sampling in 80 hospital workers (95% of those eligible). HEMA concentrations were determined in three urine samples (baseline, end of shift, and next morning) by liquid chromatography with tandem mass spectrometry. GSTs (GSTT1, GSTM1, and GSTP1) and epoxide hydrolase (EPHX1) were also genotyped. The influence of exposure, genotypes, and several other factors was examined in multiple regression analyses. Exposure was always <1 parts per million. On a group basis, exposure and a non-null GSTT1 genotype increased the HEMA concentrations in the urine sample collected at the end of the shift and these factors remained statistically significant after considering possible confounding or modifying factors. PMID- 17416774 TI - Grilled meat consumption and PhIP-DNA adducts in prostate carcinogenesis. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is the major heterocyclic amine generated from cooking meats at high temperatures, and dietary exposures have been shown to induce prostate cancer in rats. PhIP derives its carcinogenic potential through the formation of PhIP-DNA adducts. The purpose of this study was to examine whether self-reported consumption and preparation doneness of grilled meats were associated with PhIP-DNA adduct levels in prostate epithelial cells. The study population consisted of 268 African-American and Caucasian men who underwent radical prostatectomy for prostate cancer. PhIP-DNA adducts in tumor and adjacent nontumor cells were measured using immunohistochemical methods, and dietary meat intake information was based on food frequency questionnaires. Data were analyzed using multivariate linear regression models. After adjusting for age at prostatectomy and race, grilled meat consumption (P = 0.002) was significantly associated with higher adduct levels in tumor cells, but this association seemed to be primarily due to consumption of grilled red meats (P = 0.001) as opposed to grilled white meat consumption (P = 0.15). Among the specific food items, grilled hamburger consumption had the most significant association with adduct level in tumor cells (P = 0.002). Similar trends in positive associations with grilled meat consumption and adduct levels were observed in nontumor cells, but none of these associations reached statistical significance. Our results suggest that dietary interventions targeted at lower consumption of grilled red meats may reduce prostate cancer risk via the PhIP prostate carcinogenic pathway. PMID- 17416775 TI - Confirmation of a positive association between prostate cancer risk and a locus at chromosome 8q24. AB - BACKGROUND: Family-based linkage studies, association studies, and studies of tumors have highlighted human chromosome 8q as a genomic region of interest for prostate cancer susceptibility loci. Recently, a locus at 8q24, characterized by both a single nucleotide polymorphism (SNP) and a microsatellite marker, was shown to be associated with prostate cancer risk in Icelandic, Swedish, and U.S. samples. Although the data were provocative, the U.S. samples were not population based, which precludes assessment of the potential contribution of this locus to prostate cancer incidence in the United States. METHODS: We analyzed both markers in a population-based, case-control study of middle-aged men from King County, Washington. RESULTS: Overall, there was a significant positive association between the A allele of the SNP rs1447295 and prostate cancer risk [odds ratio, 1.4; 95% confidence interval (95% CI), 1.1-2.0] but no significant association with the microsatellite DG8S737. However, significant associations were observed for both markers in men with high Gleason scores. Adjusting for age, first-degree family history of prostate cancer, and prostate cancer screening history, the adjusted odds ratios were 1.4 (95% CI, 1.1-1.8) for the A allele of the SNP and 1.9 (95% CI, 1.2-2.8) for the -10 allele of the microsatellite. CONCLUSIONS: These data suggest that the locus on chromosome 8q24 harbors a genetic variant associated with prostate cancer and that the microsatellite marker is a stronger risk factor for aggressive prostate cancers defined by poorly differentiated tumor morphology. PMID- 17416776 TI - Telomere length, cigarette smoking, and bladder cancer risk in men and women. AB - Truncated telomeres are among the defining characteristics of most carcinomas. Given the role of telomeres in tumorigenesis, we reasoned that constitutionally short telomeres might be associated with an increased risk of bladder cancer. Using quantitative real-time PCR, we measured relative telomere length in bladder cancer cases and healthy controls and evaluated the association between telomere length, cigarette smoking, and bladder cancer risk in a case-control study nested within the Health Professionals Follow-up Study and a case-control study nested within the Nurses' Health Study. Telomeres were significantly shorter in bladder cancer cases (n = 184) than in controls (n = 192). The mean relative telomere length in cases was 0.23 (SD, 0.16) versus 0.27 (SD, 0.15) in controls (P = 0.001). The adjusted odds ratio for bladder cancer was 1.88 (95% confidence interval, 1.05, 3.36) for individuals in the quartile with the shortest telomeres as compared with individuals in the quartile with the longest telomeres (P(trend) = 0.006). We observed a statistically significant difference in telomere length among men and women (P < 0.001); however, the interaction between gender, telomere length, and bladder cancer risk was not significant. We also observed a significant difference in telomere length across categories of pack-years of smoking (P = 0.01). These findings suggest that truncated telomeres are associated with an increased risk of bladder cancer. PMID- 17416777 TI - Can we trust national smoking prevalence figures? Discrepancies between biochemically assessed and self-reported smoking rates in three countries. AB - BACKGROUND: National smoking prevalence estimates are the primary basis for assessing progress in tobacco control across the world. They are based on surveys of self-reported cigarette smoking. It has been assumed that this is sufficiently accurate for policy purposes, but this assumption has not been adequately tested. METHODS: We report data from the 2003 Health Survey for England, the U.S. National Health and Nutrition Examination Survey for 2001-2002, and the 2004 national smoking behaviors survey in Poland as examples of countries at different stages in the "tobacco epidemic." Self-reported cigarette and total tobacco smoking prevalence were assessed by means of the standard questions used in each country. In subsamples, specimens were collected for analysis of cotinine (saliva, N = 1,613 in England; serum, N = 4,687 in the United States; and saliva, N = 388 in Poland) providing an objective means of determining active smoking. A cut point of 15 ng/mL was used to discriminate active smoking from passive smoke exposure. RESULTS: Self-reported cigarette smoking prevalence using the standard methods underestimated true tobacco smoking prevalence by an estimated 2.8% in England, 0.6% in the United States, and 4.4% in Poland. Cotinine concentrations in those misclassified as nonsmokers were indicative of high levels of smoke intake. INTERPRETATION: Underestimation of smoking prevalence was minimal in the United States but significant in England and Poland. A review of methodologies for assessing tobacco smoking prevalence worldwide is urgently needed. PMID- 17416778 TI - The UDP-glucuronosyltransferase 2B17 gene deletion polymorphism: sex-specific association with urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol glucuronidation phenotype and risk for lung cancer. AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone is a potent and abundant procarcinogen found in tobacco smoke, and glucuronidation of its major metabolite, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), by UDP glucuronosyltransferases (UGT) including UGT2B17 is an important mechanism for 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone detoxification. Both copies of the UGT2B17 gene are deleted in approximately 10% of Whites and the deletion is associated with a reduction in NNAL glucuronidation activity in vitro. In this study, we examined the effects of the UGT2B17 deletion (0/0) on NNAL glucuronidation rates in a sample of 82 healthy cigarette smokers and further examined its effects on lung cancer risk in a separate case-control study. In the healthy smokers study, a lower urinary ratio of NNAL-glucuronide to NNAL was observed in women with the UGT2B17 deletion (0/0) as compared with women with either the wild-type or heterozygous genotypes (P = 0.058). There were no significant differences in this ratio by genotype in men (P = 0.597). In the case control study of 398 lung cancer patients and 697 community controls, the UGT2B17 deletion (0/0) was associated with a significant increase in risk of lung cancer in women (odds ratio, 2.0; 95% confidence interval, 1.01-4.0). The risk for the subset of women with lung adenocarcinoma was 2.8 (95% confidence interval, 1.2 6.3). The deletion was not associated with other lung histologic types in women and was not associated with the risk for any lung histologic types in men. The association of the UGT2B17 deletion with increased lung adenocarcinoma in women is consistent with its association with decreased NNAL glucuronidation rates in women and with studies showing that NNAL is a selective inducer of lung adenocarcinoma in experimental animals. PMID- 17416779 TI - Clinical and biological activity of soy protein powder supplementation in healthy male volunteers. AB - PURPOSE: To determine if a commonly used soy protein supplement exhibits biological activity in vivo and in vitro, we evaluated an over-the-counter soy protein powder supplement using blood from healthy male volunteers and in an estrogen receptor in vitro assay. SUBJECTS AND METHODS: We recruited healthy male volunteers 18 years of age or older that were in good health. Treatment consisted of consuming two scoops (56 g) of pure soy protein powder (Puritan's Pride, Oakdale, NY) daily for 28 days. Serum testosterone and luteinizing hormone (LH) levels were collected on days -7, 0, 14, and 28 of therapy, and day 42. A reporter estrogen receptor (ER) assay was used to determine the effect on ER-beta and ER-alpha in vitro. RESULTS: Twelve subjects were enrolled with a mean age of 32.25 years (range 25 to 47). Serum testosterone decreased 19%(+/-22%) during the 4-week use of soy protein powder (P = 0.021) and increased within 2 weeks after we discontinued soy protein powder. Serum LH concentrations decreased during the 4-week use of soy protein powder then increased within 2 weeks after we stopped the soy protein powder, but the changes did not reach statistical significance (P = 0.20). Soy protein powder was found to induce agonist activity to ER-beta using a reporter estrogen receptor assay in yeast. CONCLUSION: Soy protein powder decreases serum testosterone levels in healthy men and acts as an ER-beta agonist; the significance of this biological effect with respect to cancer prevention needs further study. PMID- 17416780 TI - Health care utilization by adult long-term survivors of hematopoietic cell transplant: report from the Bone Marrow Transplant Survivor Study. AB - The high intensity of therapy and prolonged immune suppression after hematopoietic cell transplantation (HCT) increase the risk of long-term complications and health care needs among survivors. The aim of this study was to evaluate the current status of health care utilization by long-term HCT survivors and to identify factors associated with lack of utilization. A total of 845 individuals who had undergone HCT between 1974 and 1998 at age 21 years or older and survived 2 or more years after HCT participated in the study. Health care utilization was assessed through a mailed questionnaire in three domains: general contact with health care system, general physical examination, and cancer/HCT related visit. The median age at HCT was 38.2 years, and the median length of follow-up was 6.4 years. Overall, 98% of allogeneic and 94% of autologous HCT survivors reported medical contact 11+ years after HCT. Cancer/HCT-related visits decreased with increasing time from HCT (allogeneic HCT, 98-57%; autologous HCT, 94-63%). The prevalence of general physical examination increased with time (allogeneic HCT, 56-74%; autologous HCT, 72-81%). Primary care physicians provide health care for an increasing number of adult long-term survivors of HCT, emphasizing the need for increased awareness of the long-term follow-up needs of the HCT survivors by the health care providers. PMID- 17416781 TI - Mouthwash as a low-cost and safe specimen transport medium for human papillomavirus DNA testing of cervicovaginal specimens. AB - The usefulness of mouthwash as a transport medium for cervical specimens for carcinogenic human papillomavirus (HPV) DNA testing has not been evaluated. Two cervical specimens were collected from each of 34 patients, with one placed in mouthwash (Scope, Proctor and Gamble, Inc.) and the other in a liquid cytology medium commonly used for HPV DNA testing in alternating order. Paired specimens were tested by a PCR assay for carcinogenic HPV and a PCR HPV genotyping assay for 37 HPV types at 0, 3, and 6 weeks after collection; the results of the HPV genotyping assay were categorized into HPV risk groups according to cancer risk (HPV-16 > HPV-18 > other carcinogenic HPV types > noncarcinogenic HPV types > negative). After 4 months of storage, specimens were tested using a second, non PCR test for carcinogenic HPV. We observed a >or=94% total agreement and kappa values of >or=0.88 between media at each time point for PCR-detected carcinogenic HPV. We observed a >or=74% total agreement, >or=0.62 unweighted kappa, and >or=0.75 linearly weighted kappa between media at each time point for PCR detected HPV cancer risk category. Finally, we observed an 88% total agreement and kappa of 0.77 between media for carcinogenic HPV detection using a second test after 4 months of storage. We suggest that mouthwash might be used as a low cost, safe, nonflammable storage and transport medium for cervical specimens for HPV DNA testing in cervical cancer screening programs. PMID- 17416782 TI - Glioma risk in relation to serum levels of insulin-like growth factors. AB - Several studies have suggested that insulin-like growth factors (IGF) are related to cancer risk. We investigated the associations between serum levels of IGF-I and IGF-binding protein-3 and glioma risk. A nested case-control study was conducted within a cancer prevention study, including 29,133 men (ages 50-69 years). In total, 22 glioma cases and 400 randomly selected controls were included. Serum samples were collected a minimum of 5 years before cancer diagnosis. Serum concentrations were measured using ELISA and divided into tertiles based on measurements among controls. Odds ratios and 95% confidence intervals were calculated using the lowest tertile as the reference category. No statistical association was detected between glioma and IGF-binding protein-3. IGF-I was inversely associated with glioma when comparing the lowest tertile with the other tertiles combined (odds ratio, 0.3; 95% confidence interval, 0.1-0.7). The results encourage future research on IGFs in relation to brain tumors in larger studies. PMID- 17416783 TI - Induction of the phase 2 response in mouse and human skin by sulforaphane containing broccoli sprout extracts. AB - The isothiocyanate sulforaphane was isolated from broccoli extracts in a bioactivity-guided fractionation as the principal and very potent inducer of cytoprotective phase 2 enzymes and subsequently shown to inhibit tumor development in animal models that involve various carcinogens and target organs. Because broccoli and broccoli sprouts are widely consumed, extracts obtained from them are viewed as convenient vehicles for sulforaphane delivery to humans. In relation to our current interest in devising strategies for protection against UV light-induced skin cancer, it was necessary to examine the safety and efficacy of topical application of sulforaphane-containing broccoli sprout extracts as single and multiple doses in both mice and humans. Topical application of an extract delivering 100 nmol sulforaphane/cm(2) increased the protein levels of NAD(P)H:quinone oxidoreductase 1 (NQO1), glutathione S-transferase A1, and heme oxygenase 1, three representative phase 2 enzymes, in mouse skin epidermis. Quantitative assessment of the activity of NQO1 24 h after dosing showed increases of 1.5- and 2.7-fold after application of single and multiple (thrice, every 24 h) doses, respectively. A dose-escalation safety study in healthy human subjects revealed no adverse reactions when doses as high as 340 nmol of sulforaphane in the form of broccoli sprout extracts were applied topically to the center of a 1-cm-diameter circle drawn on the volar forearm. A subsequent efficacy study showed that despite the interindividual differences in basal levels, the enzyme activity of NQO1 in homogenates of 3-mm full thickness skin punch biopsies increased in a dose-dependent manner, with maximum increases of 1.5- and 4.5-fold after application of 150 nmol doses, once or three times (at 24 h-intervals), respectively, thus providing direct evidence for induction of the phase 2 response in humans. PMID- 17416784 TI - No association of consumption of animal foods with risk of ovarian cancer. PMID- 17416785 TI - An evaluation of methods of assessing impaired awareness of hypoglycemia in type 1 diabetes. PMID- 17416786 TI - Maternal obesity and risk of gestational diabetes mellitus. AB - OBJECTIVE: Numerous studies in the U.S. and elsewhere have reported an increased risk of gestational diabetes mellitus (GDM) among women who are overweight or obese compared with lean or normal-weight women. Despite the number and overall consistency of studies reporting a higher risk of GDM with increasing weight or BMI, the magnitude of the association remains uncertain. This meta-analysis was conducted to better estimate this risk and to explore differences across studies. RESEARCH DESIGN AND METHODS: We identified studies from three sources: 1) a PubMed search of relevant articles published between January 1980 and January 2006, 2) reference lists of publications selected from the PubMed search, and 3) reference lists of review articles on obesity and maternal outcomes published between January 2000 and January 2006. We used a Bayesian model to perform the meta-analysis and meta-regression. We included cohort-designed studies that reported obesity measures reflecting pregnancy body mass, that had a normal weight comparison group, and that presented data allowing a quantitative measurement of risk. RESULTS: Twenty studies were included in the meta-analysis. The unadjusted ORs of developing GDM were 2.14 (95% CI 1.82-2.53), 3.56 (3.05 4.21), and 8.56 (5.07-16.04) among overweight, obese, and severely obese compared with normal-weight pregnant women, respectively. The meta-regression analysis found no evidence that these estimates were affected by selected study characteristics (publication date, study location, parity, type of data collection [retrospective vs. prospective], and prevalence of GDM among normal weight women). CONCLUSIONS: Our findings indicate that high maternal weight is associated with a substantially higher risk of GDM. PMID- 17416787 TI - The effects of hypogonadism on body composition and bone mineral density in type 2 diabetic patients. PMID- 17416788 TI - Visfatin response to glucose is reduced in women with gestational diabetes mellitus. PMID- 17416789 TI - Dietary patterns and the metabolic syndrome in Korean adolescents: 2001 Korean National Health and Nutrition Survey. PMID- 17416790 TI - A1C in gestational diabetes mellitus in Asian Indian women. PMID- 17416791 TI - Ferritin and transferrin are associated with metabolic syndrome abnormalities and their change over time in a general population: Data from an Epidemiological Study on the Insulin Resistance Syndrome (DESIR). AB - OBJECTIVE: The aim of this work was to study cross-sectional and longitudinal relations between iron stocks (ferritin) and the iron transport protein (transferrin) with the metabolic syndrome and its abnormalities. RESEARCH DESIGN AND METHODS: A total of 469 men and 278 premenopausal and 197 postmenopausal women from the French Data from an Epidemiological Study on the Insulin Resistance Syndrome (DESIR) cohort, aged 30-65 years, were followed over 6 years. RESULTS: Higher concentrations of both ferritin and transferrin were associated with the International Diabetes Federation (IDF) and the National Cholesterol Education Program Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults Adult Treatment Panel III original and revised definitions of the metabolic syndrome at baseline: for the IDF definition of the metabolic syndrome, the standardized, age-adjusted odds ratios (95% CI) for log(ferritin) were 1.49 (1.14-1.94) for men, 2.10 (1.27-3.48) for premenopausal women, and 1.80 (1.21-2.68) for postmenopausal women; for transferrin they were, respectively, 1.94 (1.53-2.47), 2.22 (1.32-3.75), and 2.14 (1.47-3.10). After 6 years of follow-up, the change in the presence of the metabolic syndrome was associated with higher baseline values in all three groups: log(ferritin), 1.46 (1.13-1.89), 1.28 (0.85-1.94), and 1.62 (1.10-2.38); and transferrin, 1.41 (1.10 1.81), 1.63 (1.05-2.52), and 1.51 (1.02-2.22). Among syndrome components, hypertriglyceridemia at 6 years was the component most strongly associated with baseline ferritin and transferrin. The odds of an incident IDF-defined metabolic syndrome after 6 years was more than fourfold higher when ferritin and transferrin values were both above the group-specific top tertile, in comparison with participants with both parameters below these thresholds. CONCLUSIONS: This is the first prospective study associating ferritin and transferrin with the metabolic syndrome and its components. When both markers of the iron metabolism are elevated, the incidence of the metabolic syndrome is increased in men and both pre- and postmenopausal women. PMID- 17416792 TI - Interleukin-6 in obese children and adolescents with and without glucose intolerance. PMID- 17416793 TI - Normal glucose tolerance and gestational diabetes mellitus: what is in between? AB - OBJECTIVE: The aim of this article was to define the metabolic phenotype of pregnant women with one abnormal value (OAV) during an oral glucose tolerance test (OGTT) and to test whether OAV could be considered metabolically comparable to gestational diabetes mellitus (GDM) or a specific entity between GDM and normal pregnancy. RESEARCH DESIGN AND METHODS: After 100-g 3-h OGTTs, 4,053 pregnant women were classified as having GDM, OAV, or normal glucose tolerance (NGT). Those with OAV were subdivided into three subgroups: fasting hyperglycemia (one abnormal value at fasting during an OGTT), 1-h hyperglycemia (one abnormal value at 1 h during an OGTT [1h-OAV]), or 2- or 3-h hyperglycemia (one abnormal value at 2 or 3 h during an OGTT). As derived from the OGTT, we measured insulin sensitivity (insulin sensitivity index [ISI] Matsuda) and insulin secretion (homeostasis model assessment for the estimation of beta-cell secretion [HOMA-B], first- and second-phase insulin secretion). The product of the first-phase index and the ISI was calculated to obtain the insulin secretion-sensitivity index (ISSI). RESULTS: GDM was diagnosed in 17.9% and OAV in 18.7% of pregnant women; women with GDM and OAV were older and had higher BMI and serum triglyceride levels than those with NGT (all P < 0.05). Women with NGT had the highest ISI followed by those with OAV (-21.7%) and GDM (-32.1%). HOMA-B results were comparable with those for OAV and GDM but significantly (P < 0.01) lower than those for NGT; first- and second-phase insulin secretion appeared progressively reduced from that in women with NGT to that in women with OAV and GDM (P < 0.01). ISSI was higher in women with NGT than in women with either OAV (-34%) or GDM ( 51.7%) (P < 0.001). Among OAV subgroups, the 1h-OAV subgroup showed the lowest ISSI (P < 0.05). CONCLUSIONS: OAV and GDM are clinically indistinguishable, and both groups are different from women with NGT. Women with GDM and OAV showed impaired insulin secretion and insulin sensitivity, although these defects are more pronounced in women with GDM. Compared with other OAV subgroups, 1h-OAV could be considered a more severe condition. PMID- 17416794 TI - Insulin resistance and clustering of cardiometabolic risk factors in urban teenagers in southern India. AB - OBJECTIVE: We sought to study the occurrence of cardiometabolic risk variables, their clustering, and their association with insulin resistance among healthy adolescents in urban south India. RESEARCH DESIGN AND METHODS: School children aged 12-19 years (n = 2,640; 1,323 boys and 1,317 girls) from diverse socioeconomic backgrounds were studied. Demographic, social, and medical details were obtained; anthropometry and blood pressure were measured. Fasting plasma glucose, insulin, and lipid profiles were measured. Clusters of risk variables were identified by factor analysis. Association of insulin resistance (homeostasis model assessment) with individual risk variables and their clusters were assessed. RESULTS: One or more cardiometabolic abnormalities (i.e., low HDL cholesterol, elevated triglycerides, fasting plasma glucose, or blood pressure) was present in 67.7% of children (in 64.8% of normal weight and 85% of overweight children). Insulin resistance was associated with the above abnormalities except HDL cholesterol. It also showed significant positive association with BMI, waist circumference, body fat percentage, and total cholesterol (P < 0.0001). Factor analysis identified three distinct clusters, with minor differences in the sexes: 1) waist circumference and blood pressure; 2) dyslipidemia, waist circumference, and insulin; and 3) waist circumference, glucose, and plasma insulin, with minor differences in the sexes. Insulin was a component of the lipid and glucometabolic cluster. In girls, it was a component of all three clusters. CONCLUSIONS: Cardiometabolic abnormalities are present in nearly 68% of young, healthy, Asian Indian adolescents and even among those with normal weight. Insulin resistance is associated with individual cardiometabolic factors, and plasma insulin showed association with clustering of some variables. PMID- 17416795 TI - Circulating retinol-binding protein-4, insulin sensitivity, insulin secretion, and insulin disposition index in obese and nonobese subjects. AB - OBJECTIVE: Recent investigations disclosed an upregulation of retinol-binding protein-4 (RBP4) in the adipose tissue of several insulin-resistant mouse models and increased serum RBP4 concentration in subjects with obesity and type 2 diabetes in association with insulin resistance. There is some experimental evidence that RBP4 also could been linked to insulin secretion. RESEARCH DESIGN AND METHODS: We aimed to evaluate insulin secretion, insulin sensitivity, insulin disposition index (minimal model analysis), and circulating RBP4 (enzyme-linked immunosorbent assay) in nondiabetic men with a wide range of obesity (n = 107). RESULTS: Serum RBP4 concentration was nonsignificantly different among lean, overweight, and obese subjects. Circulating RBP4 was not associated with age, BMI, waist-to-hip ratio, or metabolic parameters, including insulin sensitivity (r = -0.03, P = 0.6). On the contrary, circulating RBP4 was negatively associated with insulin secretion, especially in obese subjects (r = -0.48, P = 0.007), in whom RBP4 also was linked to insulin disposition index (r = -0.44, P = 0.01). On multiple regression analyses to predict insulin secretion (acute insulin response [AIR(g)]), insulin sensitivity was the only factor that contributed to 17% of AIR(g) variance in nonobese subjects. In obese subjects, however, RBP4 emerged as an independent factor that contributed independently to AIR(g) variance (23%). CONCLUSIONS: Our results suggest that oversecretion of RBP4 may negatively affect beta-cell function directly or by preventing the binding of transthyretin to its receptor. These mechanisms could be behind the association between increased circulating RBP4 and type 2 diabetes. RBP4 could be one signal from insulin resistant tissues that impacts on beta-cell secretion. PMID- 17416796 TI - Incretin levels and effect are markedly enhanced 1 month after Roux-en-Y gastric bypass surgery in obese patients with type 2 diabetes. AB - OBJECTIVE: Limited data on patients undergoing Roux-en-Y gastric bypass surgery (RY-GBP) suggest that an improvement in insulin secretion after surgery occurs rapidly and thus may not be wholly accounted for by weight loss. We hypothesized that in obese patients with type 2 diabetes the impaired levels and effect of incretins changed as a consequence of RY-GBP. RESEARCH DESIGN AND METHODS: Incretin (gastric inhibitory peptide [GIP] and glucagon-like peptide-1 [GLP-1]) levels and their effect on insulin secretion were measured before and 1 month after RY-GBP in eight obese women with type 2 diabetes and in seven obese nondiabetic control subjects. The incretin effect was measured as the difference in insulin secretion (area under the curve [AUC]) in response to an oral glucose tolerance test (OGTT) and to an isoglycemic intravenous glucose test. RESULTS: Fasting and stimulated levels of GLP-1 and GIP were not different between control subjects and patients with type 2 diabetes before the surgery. One month after RY GBP, body weight decreased by 9.2 +/- 7.0 kg, oral glucose-stimulated GLP-1 (AUC) and GIP peak levels increased significantly by 24.3 +/- 7.9 pmol x l(-1) x min( 1) (P < 0.0001) and 131 +/- 85 pg/ml (P = 0.007), respectively. The blunted incretin effect markedly increased from 7.6 +/- 28.7 to 42.5 +/- 11.3 (P = 0.005) after RY-GBP, at which it time was not different from that for the control subjects (53.6 +/- 23.5%, P = 0.284). CONCLUSIONS: These data suggest that early after RY-GBP, greater GLP-1 and GIP release could be a potential mediator of improved insulin secretion. PMID- 17416797 TI - TCF7L2 polymorphisms modulate proinsulin levels and beta-cell function in a British Europid population. AB - Rapidly accumulating evidence shows that common T-cell transcription factor (TCF)7L2 polymorphisms confer risk of type 2 diabetes through unknown mechanisms. We examined the association between four TCF7L2 single nucleotide polymorphisms (SNPs), including rs7903146, and measures of insulin sensitivity and insulin secretion in 1,697 Europid men and women of the population-based MRC (Medical Research Council)-Ely study. The T-(minor) allele of rs7903146 was strongly and positively associated with fasting proinsulin (P = 4.55 x 10(-9)) and 32,33 split proinsulin (P = 1.72 x 10(-4)) relative to total insulin levels; i.e., differences between T/T and C/C homozygotes amounted to 21.9 and 18.4% respectively. Notably, the insulin-to-glucose ratio (IGR) at 30-min oral glucose tolerance test (OGTT), a frequently used surrogate of first-phase insulin secretion, was not associated with the TCF7L2 SNP (P > 0.7). However, the insulin response (IGR) at 60-min OGTT was significantly lower in T-allele carriers (P = 3.5 x 10(-3)). The T-allele was also associated with higher A1C concentrations (P = 1.2 x 10(-2)) and reduced beta-cell function, assessed by homeostasis model assessment of beta-cell function (P = 2.8 x 10(-2)). Similar results were obtained for the other TCF7L2 SNPs. Of note, both major genes involved in proinsulin processing (PC1, PC2) contain TCF-binding sites in their promoters. Our findings suggest that the TCF7L2 risk allele may predispose to type 2 diabetes by impairing beta-cell proinsulin processing. The risk allele increases proinsulin levels and diminishes the 60-min but not 30-min insulin response during OGTT. The strong association between the TCF7L2 risk allele and fasting proinsulin but not insulin levels is notable, as, in this unselected and largely normoglycemic population, external influences on beta-cell stress are unlikely to be major factors influencing the efficiency of proinsulin processing. PMID- 17416798 TI - Hypoglycemic action of thiazolidinediones/peroxisome proliferator-activated receptor gamma by inhibition of the c-Jun NH2-terminal kinase pathway. AB - Type 2 diabetes results from progressive pancreatic beta-cell dysfunction caused by chronic insulin resistance. Activation of c-Jun NH2-terminal kinase (JNK) inhibits insulin signaling in cultured cells and in vivo and thereby promotes insulin resistance. Conversely, the peroxisome proliferator-activated receptor (PPAR) gamma synthetic ligands thiazolidinediones (TZDs) enhance insulin sensitivity. Here, we show that the TZDs rosiglitazone and troglitazone inhibit tumor necrosis factor-alpha-induced JNK activation in 3T3-L1 adipocytes. Our results indicate that PPARgamma mediates this inhibitory action because 1) it is reproduced by other chemically unrelated PPARgamma agonist ligands and blocked by PPARgamma antagonists; 2) it is enhanced by PPARgamma overexpression; and 3) it is abrogated by PPARgamma RNA interference. In addition, we show that rosiglitazone inhibits JNK activation and promotes the survival of pancreatic beta-cells exposed to interleukin-1beta. In vivo, the abnormally elevated JNK activity is inhibited in peripheral tissues by rosiglitazone in two distinct murine models of obesity. Moreover, rosiglitazone fails to enhance insulin induced glucose uptake in primary adipocytes from ob/ob JNK1-/- mice. Accordingly, we demonstrate that the hypoglycemic action of rosiglitazone is abrogated in the diet-induced obese JNK1-deficient mice. In summary, we describe a novel mechanism based on targeting the JNK signaling pathway, which is involved in the hypoglycemic and potentially in the pancreatic beta-cell protective actions of TZDs/PPARgamma. PMID- 17416799 TI - Variations in adiponectin receptor genes and susceptibility to type 2 diabetes in women: a tagging-single nucleotide polymorphism haplotype analysis. AB - Adiponectin has been associated with low diabetes risk. The metabolic effects of adiponectin are mediated by adiponectin receptors 1 (ADIPOR1) and 2 (ADIPOR2). We conducted a prospective, nested case-control study of 714 cases of type 2 diabetes and 1,120 control subjects. Six polymorphisms in ADIPOR1 and 16 polymorphisms in ADIPOR2 were determined. Haplotypes inferred from ADIPOR1 polymorphisms were significantly associated with diabetes risk (overall test, 2log-likelihood = 15.1 on 5 df; P = 0.0098). A single copy of haplotype 001100 (0, common allele; and 1, minor allele) was associated with 24% decreased risk (odds ratio [OR] 0.76 [95% CI 0.61-0.96], P = 0.02) compared with the most common haplotype, 110000, adjusting for age, BMI, and other covariates. A 3' untranslated region (UTR) polymorphism, rs1139646, showed the strongest and nominally significant association with greater diabetes risk (unadjusted OR 1.26 [1.03-1.53] and adjusted OR 1.36 [1.10-1.70]). However, such an association became marginal after controlling for multiple comparisons by permutation test (P = 0.08 on the basis of 10,000 permutations). There were not significant associations between ADIPOR2 polymorphisms, individually or in haplotypes, and the risk of type 2 diabetes. In conclusion, our data indicate significant associations between ADIPOR1 haplotypes and diabetes risk but do not support a relation between ADIPOR2 variability and the disease. PMID- 17416800 TI - Quantitative trait loci for fasting glucose in young Europeans replicate previous findings for type 2 diabetes in 2q23-24 and other locations. AB - Long before reaching diagnostic cutoff levels for type 2 diabetes, fasting glucose can be a powerful risk marker for this disease. We conducted a genome wide search for fasting glucose as a quantitative trait in 412 young European sib pairs including obese children, with adjustment for sex, age, and BMI. We identified more quantitative trait loci specific to fasting glucose and more significant than would be found by simple chance estimated by permutation tests. The strongest linkage was on chromosome 2q (logarithm of odds [LOD] = 3.00) in a region previously linked to type 2 diabetes as a disease. We also found linkage signals of fasting glucose with 7q (LOD = 2.03), 8q (1.28), 17p (2.12), 17q (1.4), and 11p (1.33). These findings suggest that the quantitative genetics of fasting glucose could contribute to the search for type 2 diabetes genes. PMID- 17416801 TI - Triglyceride uptake and lipoprotein lipase-generated fatty acid spillover in the splanchnic bed of dogs. AB - The action of lipoprotein lipase on triglyceride-rich lipoproteins generates fatty acids that are either transported into tissues or mix with circulating free fatty acids (FFAs) via a process known as spillover. In the present study, arterial, portal vein, and hepatic vein sampling catheters were surgically placed in nine mongrel dogs. The animals were subsequently studied after a 42-h fast during infusion of [14C]oleate and a lipid emulsion containing [3H]triolein; the emulsion was used as a surrogate for the study of chylomicron metabolism. More than one-half of splanchnic [3H]triglyceride uptake occurred in the liver, and substantial fractional spillover of [3H]oleate was observed in both liver and nonhepatic tissues (approximately 50% each). There was a significant correlation between FFA release from nonhepatic tissues (presumably visceral fat) and nonhepatic fractional spillover (R = 0.81, P < 0.01), consistent with a model in which the rate of intracellular lipolysis influences spillover by determining the direction of net fatty acid flow between the cell and the interstitium. There was a significant correlation between "true" and "net" splanchnic spillover (R = 0.84, P < 0.005), the latter representing calculation of spillover between arterial and hepatic venous blood without portal venous data. Metabolism of chylomicron triglycerides in visceral fat may be an important source of portal venous FFAs. PMID- 17416802 TI - How American is the American Journal of Neuroradiology? PMID- 17416803 TI - Presence of activated microglia in a high-signal lesion on T1-weighted MR images: a biopsy sample re-examined. PMID- 17416804 TI - The Neuroimaging Center of the Pediatric Brain Tumor Consortium-collaborative neuroimaging in pediatric brain tumor research: a work in progress. AB - As an essential part of the National Cancer Institute (NCI)-funded Pediatric Brain Tumor Consortium (PBTC), the Neuroimaging Center (NIC) is dedicated to infusing the study of pediatric brain tumors with imaging "best practice" by producing a correlative research plan that 1) resonates with novel therapeutic interventions being developed by the wider PBTC, 2) ensures that every PBTC protocol incorporates an imaging "end point" among its objectives, 3) promotes the widespread implementation of standardized technical protocols for neuroimaging, and 4) facilitates a quality assurance program that complies with the highest standards for image data transfer, diagnostic image quality, and data integrity. To accomplish these specific objectives, the NIC works with the various PBTC sites (10 in all, plus NCI/ National Institute of Neurological Diseases and Stroke representation) to ensure that the overarching mission of the consortium--to better understand tumor biology and develop new therapies for central nervous system tumors in children--is furthered by creating a uniform body of imaging techniques, technical protocols, and standards. Since the inception of the NIC in 2003, this broader mandate has been largely accomplished through a series of site visits and meetings aimed at assessing prevailing neuroimaging practices against NIC-recommended protocols, techniques, and strategies for achieving superior image quality and executing the secure transfer of data to the central PBTC. These ongoing evaluations periodically examine investigations into targeted drug therapies. In the future, the NIC will concentrate its efforts on improving image analysis for MR imaging and positron emission tomography (PET) and on developing new ligands for PET; imaging markers for radiation therapy; and novel systemic, intrathecal, and intralesional therapeutic interventions. PMID- 17416805 TI - Unique imaging findings in the facial bones of renal osteodystrophy. AB - Facial skeletal changes associated with hyperparathyroidism assume 3 radiographic patterns: osteitis fibrosa cystica, fibrous dysplasia, and leontiasis ossea. The 3rd pattern is unique to renal osteodystrophy. We report a case of uremic leontiasis ossea with CT images illustrating significant hypertrophy of the jaws with serpiginous tunneling within the bone and poor visualization of the cortical bone. PMID- 17416806 TI - Arachnoid granulations of the posterior temporal bone wall: imaging appearance and differential diagnosis. AB - Arachnoid granulations are rarely seen on high-resolution CT (HRCT) at the posterior temporal bone wall, where they appear as erosions, without bone spicules and often with a lobulated surface. Differential diagnosis includes endolymphatic sac tumor, paraganglioma, chordoma, and chondromatous and metastatic tumors. MR imaging can confirm the diagnosis because arachnoid granulations behave like CSF without gadolinium enhancement. This report aims to illustrate the appearance and differentiation of temporal bone arachnoid granulations on HRCT and MR imaging. PMID- 17416807 TI - Spinal metastasis from endolymphatic sac tumor. AB - We describe a case of endolymphatic sac tumor with drop metastasis to the spine. Our review of the literature showed that this is only the 2nd reported case of such an occurrence. PMID- 17416808 TI - Intraosseous lipoma of the left frontoethmoidal sinuses and nasal cavity. AB - Intraosseous lipomas are very rare slow-growing benign tumors that may appear as congenital lesions or may be acquired (metaplasia from a pre-existing lipoma). Only a handful of head and neck cases have been reported in the literature. We present the first reported case of a solitary osteolipoma involving the sinonasal tract in a 66-year-old man with sinonasal symptoms. CT showed a lesion involving the left frontal sinus, extending into the ethmoid sinus with insinuation along the left middle turbinate. It appeared to be an atypical mass with areas of high attenuation (calcification) within. PMID- 17416809 TI - Focal hyperostosis on CT of sinonasal inverted papilloma as a predictor of tumor origin. AB - BACKGROUND AND PURPOSE: To evaluate the CT characteristics of focal hyperostosis in patients with sinonasal inverted papilloma and to correlate these characteristics with the detection of the origins of tumors. MATERIALS AND METHODS: Paranasal sinus and nasal cavity CT images of 76 patients were reviewed retrospectively to detect areas within which there was focal hyperostosis. We correlated the sites on the CT scans within which there was focal hyperostosis with the origin of the tumors described in the corresponding patient's medical records. We also evaluated the CT features of focal hyperostosis according to the origin of tumors. RESULT: Surgical evaluation of 55 lesions with focal hyperostosis in CT images revealed that 49 of these lesions coincided with the actual origin of tumor. The CT-based determination of the locations of the areas of focal hyperostosis corresponded to the actual tumor origin in 89.1% of cases. Especially in cases with focal hyperostosis within the frontal, maxillary, sphenoid, and posterior ethmoid sinuses, areas of focal hyperostosis corresponded to the origin of tumor without exception. In the evaluation of the CT features of focal hyperostosis, 2 patterns of localized bone thickening were noted. Plaquelike bone thickening was seen mainly when focal hyperostosis involved the lateral wall of the nasal cavity. On the other hand, cone-shaped bone thickening was seen only in the walls of the paranasal sinuses or the bony septum. CONCLUSIONS: A high correlation between the origin of the inverted papilloma and focal hyperostosis on CT might facilitate preoperative prediction of tumor origin by radiologists and rhinologists. PMID- 17416810 TI - Intra-aneurysmal hemodynamics during the growth of an unruptured aneurysm: in vitro study using longitudinal CT angiogram database. AB - BACKGROUND AND PURPOSE: The role of blood-flow biomechanics on the size, morphology, and growth of cerebral aneurysms is poorly known. The purpose of this study was to evaluate intra-aneurysmal hemodynamics before and after aneurysm growth. MATERIALS AND METHODS: A flow-simulation study was performed in a middle cerebral artery (MCA) aneurysm with a bleb that grew after 1-year follow-up. Geometrically realistic in vitro models before and after aneurysm growth were constructed on the basis of CT angiograms. Blood-flow velocity, vorticity, and wall shear stress were obtained by using particle imaging velocimetry and laser Doppler velocimetry. RESULTS: No significant quantitative differences were noted among the overall flow structures before and after aneurysm growth, with the exception of less vorticity in the bleb after aneurysm growth. A circulating flow pattern was seen within the aneurysm domes. A blood-flow separation was observed at the margins of the bleb. No impingement of inward flow into the enlarging bleb was noted. Before the aneurysm growth, the wall shear stress was high at the aneurysm neck and also at the margin of the bleb. The value of wall shear stress decreased in the deeper part of the bleb. This value decreased even more after the aneurysm growth. CONCLUSIONS: Intra-aneurysmal hemodynamic structures before and after the growth of an MCA aneurysm were compared. Further investigation with a similar approach is mandatory to obtain a firm conclusion. PMID- 17416811 TI - Diagnostic accuracy of 3D time-of-flight MR angiography compared with digital subtraction angiography for follow-up of coiled intracranial aneurysms: influence of aneurysm size. AB - BACKGROUND AND PURPOSE: 3D time-of-flight MR angiography (3D TOF MRA) may be used as noninvasive alternative to digital subtraction angiography (DSA) for the follow-up of patients with intracranial aneurysms treated with Guglielmi detachable coils (GDCs). We aimed to determine the influence of aneurysm size and location on diagnostic accuracy of 3D TOF MRA for follow-up of intracranial aneurysms treated with GDCs. MATERIALS AND METHODS: Two hundred and one 3D TOF MRAs in 127 consecutive patients with 136 aneurysms were compared with DSA as standard of reference. Sensitivity and specificity of 3D TOF MRA for detection of residual or reperfusion of the aneurysms was calculated with regard to aneurysm size and location. RESULTS: Overall sensitivity and specificity of MRA was 88.5% and 92.9%, respectively. Sensitivity was lower for aneurysms or=70% internal carotid artery stenosis. Transcranial Doppler (TCD) was done before, 6 hours, and 30 days after CAS. Cerebrovascular reserve to apnea (CVR) was measured. RESULTS: On the ipsilateral side, basal middle cerebral artery (MCA) middle flow velocity (MFV) and pulsatility index (PI) increased from 49.7 cm/s and 0.85, respectively, to 62.5 cm/s and 1.09 immediately after CAS (P < .05); anterior cerebral artery (ACA) MFV and PI increased from 50.7 cm/s and 0.90, to 58.9 cm/s and 1.08 (P < .05); and posterior cerebral artery (PCA) MFV decreased from 47.5 to 36.5 cm/s, with no change in PI (P < .05). On the contralateral side, nonsignificant changes were seen on MCA, immediately after CAS; ACA and PCA MFV decreased from 63.7 and 45.3 cm/s to 50.3 and 38.6 cm/s, respectively (P < .05); ACA and PCA PI increased from 0.96 and 1.00, respectively, to 1.04 and 1.04 (P < .05). At 30 days, ipsilateral MCA MFV and PI were 52.8 cm/s and 1.12, respectively (P < .05), and contralateral values were 49.6 cm/s (P < .05) and 1.02 (nonsignificant), respectively. Basal ipsilateral and contralateral CVR improved from 26.0% to 37.0% (P < .05), and from 30.1% to 33.5% (nonsignificant), respectively, at 30 days. CONCLUSIONS: CAS produces an early significant increase of MFV and PI in the ipsilateral anterior circulation. This effect is maintained, though minor, 30 days later. In addition, CAS results in an improvement of CVR at 30 days. PMID- 17416814 TI - Reduction of radiation dose for cerebral angiography using flat panel detector of direct conversion type: a vascular phantom study. AB - BACKGROUND AND PURPOSE: Compared with image intensifier television (I.I.-TV) system, an angiography system using the flat panel detector (FPD) of direct conversion type has a high spatial resolution, which may improve image quality, reduce patient exposure, or both. Our purpose was to evaluate the detection of simulated aneurysmal blebs under dose reduction with the FPD system in comparison with the I.I.-TV system. MATERIALS AND METHODS: A vascular phantom was designed to simulate various intracranial aneurysms with and without blebs, and this phantom was filled with 3 different concentrations of contrast material (300, 150, and 100 mg I/mL). 2D digital subtraction angiography (DSA) at low-dose mode of FPD system was compared with 2D DSA at a standard-dose mode of FPD system and a conventional mode of I.I.-TV system. Data analysis was based on 171 observations (57 aneurysms [20 with bleb and 37 without bleb] x 3 contrast material concentrations) by each of 7 radiologists, and the detection performances of blebs were compared using a receiver operating characteristic (ROC) analysis. RESULTS: The mean dose measurements with a phantom during 2D DSA were 0.36 mGy/frame with low-dose mode of FPD system, 0.72 mGy/frame with standard-dose mode of FPD system and 0.76 mGy/frame with I.I.-TV system. The mean Az at 100 mg I/mL was significantly higher for low-dose mode of FPD than for conventional-dose mode of I.I.-TV mean Az, 0.85 versus 0.56; P < .01), though differences were not significant with 150 and 300 mg I/mL between both systems. CONCLUSION: The FPD system allows a considerable dose reduction during 2D DSA without loss of the image quality. PMID- 17416815 TI - Dural arteriovenous fistulas with cortical venous drainage: incidence, clinical presentation, and treatment. AB - BACKGROUND AND PURPOSE: Our purpose was to report our experience with intracranial dural arteriovenous fistulas (DAVFs) with cortical venous drainage during a 12-year period. PATIENTS AND METHODS: Between January 1994 and January 2006, 91 patients with intracranial DAVFs presented at our institution, and 29 (32%) had cortical venous drainage. There were 5 women and 24 men (mean age, 53.9 years; range, 24-77). Clinical presentation was intraparenchymal or subarachnoid hemorrhage in 18 patients (62%), seizures in 4 patients (14%), visual symptoms in 2 patients (7%), pulsatile bruit in 1 patient (3%), and the DAVF (14%) was incidentally discovered in 4 patients. RESULTS: In 2 patients, the DAVF had been obliterated spontaneously at the time of scheduled embolization 10 and 2 months after hemorrhage, respectively. Five patients with an anterior fossa DAVF underwent successful surgery. In 14 patients, the DAVF was completely occluded with embolization alone, and in 7 patients, embolization was followed by surgery. Altogether, complete occlusion was angiographically confirmed in 28 of 29 DAVFs; the result of radiosurgery of 1 DAVF is pending. There were no complications of surgery; embolization was complicated by postembolization hemorrhage in 1 patient (3%). CONCLUSION: Most DAVFs with cortical venous drainage have an aggressive clinical course. Treatment by a neurovascular team by using surgery, embolization, or a combination resulted in cure in all cases, with a very low complication rate. PMID- 17416816 TI - Reversibility of venous sinus obstruction in idiopathic intracranial hypertension. AB - Elevated intracranial intravenous pressure seems to be of importance in pseudotumor cerebri syndromes, either as a cause (secondary intracranial hypertension) or as a consequence (idiopathic intracranial hypertension) of increased intracranial pressure. We present 3 case reports in which diagnostic imaging before and after CSF diversion provided evidence that narrowing of the transverse sinuses is a secondary phenomenon. Stent angioplasty of the venous sinuses should not be considered a therapeutic approach in these cases. PMID- 17416817 TI - Differentiating normal myelination from hypoxic-ischemic encephalopathy on T1 weighted MR Images: a new approach. AB - BACKGROUND AND PURPOSE: Hypoxic-ischemic cerebral changes can be difficult to distinguish from normal myelination on T1-weighted images. We hypothesized that comparing signal intensity (SI) of brain structures on T1-weighted images enables differentiation of myelination from hypoxic-ischemic brain damage. MATERIALS AND METHODS: T1-weighted images, obtained in 57 infants aged 1-104 days and born after a gestational age of 35 weeks or older, were retrospectively evaluated. Subjects were assigned to a patient (n = 23, with perinatal hypoxic-ischemic encephalopathy [HIE] stage 2/3) or a control group (n = 34). In each subject, an SI score was assigned to 19 brain structures on the basis of pairwise comparisons with the other 18 structures. In both groups, mean total SI scores were calculated for the 19 structures. Independent samples t tests assessed whether the mean total score of a structure differed significantly between the 2 groups. Logistic regression assessed which comparison was best to distinguish between the groups and to predict the presence of hypoxic-ischemic injury. RESULTS: In patients, mean total SI scores for posterolateral putamen (PP) and peri-Rolandic cortex (PC) were significantly higher (P = .000 for both). Mean total SI scores of the posterior limb of internal capsule (PLIC) and the corona radiata (CR) were significantly lower in patients (P = .000 and 0.005, respectively). Two comparisons (PLIC versus CR, PP versus PC) were best to distinguish patients and controls and to predict absence or presence of HIE (P < .0001). CONCLUSION: SI changes due to hypoxia-ischemia can be differentiated from normal myelination by comparing SI of 4 brain structures on T1-weighted images. PMID- 17416818 TI - Defining optic nerve tortuosity. AB - BACKGROUND AND PURPOSE: Optic nerve tortuosity is one of several nonmalignant abnormalities documented on MR imaging in patients with neurofibromatosis type 1 and may be related to the development of optic pathway gliomas. This study seeks an operational definition for optic nerve tortuosity. MATERIALS AND METHODS: A focus group of 3 pediatric neuroradiologists reviewed 20 MR images of the brain and orbits of patients suspected to have optic nerve tortuosity in the absence of optic pathway glioma and found 6 radiographic factors that occurred frequently. Subsequently, 28 MR images were assessed for the presence of optic nerve tortuosity, using a global assessment question that reflects a neuroradiologist's confidence in the presence of optic nerve tortuosity, and for the presence of the 6 radiographic factors, to identify a combination of these factors that best predicted a diagnosis of optic nerve tortuosity. RESULTS: We found perfect inter rater agreement between 3 readers on the presence/absence of tortuosity in 75% of cases. Lack of congruity of the optic nerves, in more than 1 coronal section and dilation of the subarachnoid space surrounding the optic nerves, when found together are sensitive (89%) and specific (93%) for a diagnosis of tortuosity on the global scale. The absence of these 2 factors, along with absence of deviation of the optic nerve within the axial plane, provides a reliable test to exclude tortuosity. CONCLUSION: Lack of congruity of the optic nerves in more than 1 coronal section and dilation of the subarachnoid space surrounding the optic nerves together provide an operational radiographic definition of optic nerve tortuosity. PMID- 17416819 TI - Shape mapping of the hippocampus in young children with autism spectrum disorder. AB - BACKGROUND AND PURPOSE: We hypothesized the occurrence of characteristic hippocampal-shape alterations in young children with autistic spectrum disorder (ASD) who also exhibit deficits on neuropsychologic tests of medial temporal lobe (MTL) function. MATERIALS AND METHODS: Coronal 3D MR images were acquired from 3- to 4-year-old children with ASD (n = 45) and age-matched children with typical development (n = 13). Children with ASD were further subclassified into those with autism disorder (AD, n = 29) or pervasive developmental disorder-not otherwise specified (PDD-NOS) (n = 16). Variations in hippocampal shape were evaluated by using large-deformation high-dimensional brain mapping. RESULTS: Hippocampal shape measures distinguished children with ASD from those with typical development; within the ASD sample, children with AD were distinguished from those with PDD-NOS. Hippocampal-shape alterations in children with ASD were correlated with degree of mental retardation and performance deficits on tests of MTL function. CONCLUSIONS: Children with ASD exhibited an alteration of hippocampal shape consistent with inward deformation of the subiculum. This pattern of hippocampal-shape deformations in the children with ASD was accentuated in the more severely affected subgroup of children with AD and was associated with deficits on neuropsychologic tests of MTL but not prefrontal function. Hippocampal-shape deformation in the children with ASD was observed to be similar to a pattern of hippocampal shape deformation previously reported in adults with MTL epilepsy. Although the children with ASD, and those with AD in particular, PDD-NOS are at high risk for epilepsy as they enter adolescence, the specificity and causal relationship of this pattern of hippocampal-shape deformation to the development of seizures is not yet known. PMID- 17416820 TI - Hemimegalencephaly: a study of abnormalities occurring outside the involved hemisphere. AB - BACKGROUND AND PURPOSE: Hemimegalencephaly is a rare but well-known congenital malformation with ipsilateral enlargement of the hemicerebrum. However, very little is known about changes in structures outside the involved hemisphere in patients with this condition. We investigated morphologic abnormalities occurring outside the affected hemisphere by MR imaging in a large series of patients with hemimegalencephaly. MATERIALS AND METHODS: MR imaging findings for 30 patients with hemimegalencephaly were retrospectively reviewed and evaluated for structures outside the involved hemisphere on routine MR images, such as cranial nerves (I, II, V), brain vessels, subdural and subarachnoid spaces, brain stem, and cerebellum, on both the ipsilateral and contralateral sides. RESULTS: The ipsilateral olfactory and optic nerves were enlarged in 8 (26.7%) and 1 (3.3%) of the 30 patients, respectively, without enlargement on the contralateral side. No asymmetry was noted in the trigeminal nerves. Asymmetric vascular dilations in the ipsilateral cerebral hemisphere were observed in 12 of the 30 patients (40%), in deep cerebral vessels in 11 patients (36.7%), and in superficial cerebral vessels in 8 patients (26.7%). Ipsilateral brain stem and hemicerebellar asymmetric enlargement was detected in 2 patients (6.7%) and 14 patients (46.7%), respectively. Abnormal cerebellar folia were observed on the ipsilateral side in 6 patients (20%) and on the contralateral side in 3 patients (10%). CONCLUSION: Ipsilateral olfactory nerve enlargement, cerebral vascular dilations, cerebellar enlargement, and bilateral or ipsilateral abnormal architecture of the cerebellar folia are often associated with hemimegalencephaly. PMID- 17416821 TI - Vertebroplasty, first 1000 levels of a single center: evaluation of the outcomes and complications. AB - BACKGROUND AND PURPOSE: Percutaneous polymethylmethacrylate (PMMA) vertebroplasty has become a common procedure for treatment of pain and disability associated with vertebral compression fractures. We reviewed the experience with our first 1000 consecutively treated vertebral compression fractures in an attempt to demonstrate both the short- and long-term safety and efficacy of percutaneous vertebroplasty. MATERIALS AND METHODS: The first 1000 compression fractures treated by vertebroplasty at our institution were identified from a comprehensive prospectively acquired vertebroplasty data base. All patients treated with vertebroplasty were included, regardless of the underlying pathologic cause. Chart reviews of the procedure notes, imaging studies, clinical visits, and follow-up telephone interviews were performed for each patient. Evaluation at each follow-up time point included pain response (subjective and visual analog pain score), change in mobility, change in pain medication usage, and modified Roland-Morris Disability Questionnaire. Statistical analysis was performed on the pain response and change in the Roland-Morris score at each follow-up time point. Significant procedure-related complications that occurred from the time of the procedure were also specifically extracted from the patients' charts. RESULTS: There was a dramatic improvement in all the evaluated parameters following percutaneous vertebroplasty. The improvement in pain, mobility, medication usage, and Roland-Morris score was noticed immediately after the procedure and persisted through the 2-year follow-up. There was a low rate of complications from the procedure, the most common being rib fractures. CONCLUSION: According to our results, practitioners can quote a high success rate and low complication rate for vertebroplasty when making treatment recommendations for painful spinal compression fractures. PMID- 17416822 TI - Vertebroplasty for osteoporotic fractures with spinal canal compromise. AB - BACKGROUND AND PURPOSE: Percutaneous vertebroplasty can aggravate spinal canal narrowing, especially in patients with preoperative retropulsion. The purpose of this study was to evaluate changes in spinal canal dimension during percutaneous vertebroplasty for osteoporotic fractures with retropulsion. MATERIALS AND METHODS: We reviewed all cases of osteoporotic vertebral fractures treated with vertebroplasty. Twenty-one patients (25 vertebrae) had a retropulsed fragment that compromised the dimension of the spinal canal on preoperative imaging. We measured the degree of retropulsion before and after vertebroplasty to evaluate changes in spinal canal dimension. We also evaluated pain, neurologic status, vertebral body height, and wedge angle. RESULTS: Mean retropulsion was 4.2 mm before and 4.4 mm after vertebroplasty. There was no statistically significant difference (P = .32). Mean increase in vertebral body height was 2.6 mm anteriorly, 1.7 mm centrally, and 0.3 mm posteriorly. Mean decrease in wedge angle was 4.7 degrees. There were statistically significant improvements in height and wedge angle (P < .01). None of our patients developed new symptoms during vertebroplasty or thereafter. Twenty of 21 patients (95%) showed partial or complete immediate pain relief, whereas 1 patient did not improve. CONCLUSION: Vertebroplasty can be performed safely in patients with spinal canal compromise. This procedure can reduce pain, increase vertebral body height, and decrease wedge angle without worsening of retropulsion. PMID- 17416823 TI - Follow-up MR imaging in patients with pyogenic spine infections: lack of correlation with clinical features. AB - BACKGROUND AND PURPOSE: Follow-up MR imaging examinations are increasingly used to monitor response to treatment in patients with spine infection. We aim to describe follow-up MR imaging examination findings 4-8 weeks after diagnosis and initiation of treatment of spine infections and to compare with clinical findings. MATERIALS AND METHODS: Thirty-three patients with spinal infection and available baseline and 4-8-week follow-up MRIs were included in this retrospective cohort study. Baseline and follow-up MR imaging were graded by 2 neuroradiologists blinded to clinical characteristics and outcome. Clinical findings and outcomes were independently obtained by retrospective review of the medical record. RESULTS: Compared with baseline MR imaging examinations, follow up MR imaging more frequently demonstrated vertebral body loss of height (26/33 [79%] versus 14/33 [47%]; P < .001) and less frequently demonstrated epidural enhancement (19/32 [59%] versus 29/33 [88%]; P = .008), epidural canal abscess (3/32 [9%] versus 15/33 [45%]; P = .001), and epidural canal compromise (10/32 [31%] versus 19/33 [58%]; P = .008). Most follow-up MR imaging examinations demonstrated less paraspinal inflammation and less epidural enhancement compared with baseline. However, vertebral body enhancement, disk space enhancement, and bone marrow edema more often were equivocal or appeared worse compared with baseline. Twenty-one of 32 (66%) follow-up MR imaging examination overall grades were considered improved, 5 (16%) were equivocal, and 6 (19%) were worse. No single MR imaging finding was associated with clinical status. CONCLUSION: Soft tissue findings, not bony findings, should be the focus of clinicians interpreting follow-up MR imaging results. No single MR imaging parameter was associated with the patients' clinical status. PMID- 17416824 TI - Plasma-mediated radiofrequency ablation assisted percutaneous cement injection for treating advanced malignant vertebral compression fractures. AB - BACKGROUND AND PURPOSE: Using percutaneous cement injection to treat vertebral compression fractures (VCF) stemming from advanced malignancy, particularly those showing posterior cortical defect and epidural extension, is associated with higher risk of complications compared with treating benign osteoporotic VCF. The purpose of this study was to examine the clinical feasibility of a technique designed to improve control of cement placement. MATERIALS AND METHODS: A prospective series of patients (n = 15) with metastatic lesions having epidural extension of tumor and/or cortical disruption were treated. The percutaneous procedure consisted of using a plasma-mediated radio-frequency-based device to etch a cavity within the affected vertebral body and filling the cavity and adjacent interstices with bone cement. Change in pain status was determined by asking the patient to grade back pain preoperatively and at the 2-4-week postprocedure examination using a visual analog scale. RESULTS: An adequate amount of cement was injected in all cases. Extraosseous extension of cement was observed in 4 cases (anterior cortex, n = 3; through neural foramen, n = 1) but was clinically inconsequential. No thermal or neuronal insult was observed during the procedure in any case. Thirteen of the 15 (87%) patients reported decreased pain. CONCLUSION: Dissolution of tissue rather than displacement to create a cavity before injecting bone cement permitted well-directed cement deposition into the compromised vertebral body, which may allow a safer procedure to be conducted in patients with advanced malignant VCF. Clinical benefits may include avoiding more extensive surgery and reducing the risk of complications associated with conventional bone cement injection procedures. PMID- 17416825 TI - Lower cervical nerve root block using CT fluoroscopy in patients with large body habitus: another benefit of the swimmer's position. AB - We describe a method of performing lower cervical nerve root block (CNRB) with CT fluoroscopy in patients with large body habitus using the swimmer's position. This approach reduces image noise with acceptable visualization of vital structures and improved foraminal/root access. Anticipated use of the swimmer's position coupled with minimally modified radiation exposure parameters can limit radiation dose to operator/patient and reduce procedure time to match that of CNRB using CT fluoroscopy in typical patients. PMID- 17416826 TI - MR imaging findings of paravertebral gossypiboma. AB - BACKGROUND AND PURPOSE: "Gossypiboma" is a term used to describe a mass within the body that comprises a cotton matrix surrounded by a foreign-body reaction, which is extremely rare after spinal surgery. The purpose of this study was to describe the clinical and MR imaging features of gossypibomas in the paravertebral area of 7 patients. MATERIALS AND METHODS: Between October 1999 and December 2005, 7 cases of paravertebral gossypibomas were confirmed pathologically at 2 institutions. The clinical features were reviewed retrospectively. We recorded MR imaging features and differential MR imaging based diagnoses. RESULTS: All patients were symptomatic; the most frequent symptom was nonspecific back or neck pain. None of the patients presented with signs of infection or required surgery urgently. In 3 of the 7 patients, gossypiboma was included in the preoperative differential imaging diagnosis. In all patients, the gossypiboma was located in the vicinity of the initial site of surgery. Typically, gossypibomas were visible in T2-weighted images as a mass with a hyperintense center and hypointense rim and exhibited strong peripheral enhancement in contrast-enhanced images. Retained surgical gauze was removed surgically from 4 patients; in the remaining patients, the surgical specimens contained suture threads. CONCLUSION: Gossypiboma should be included in the differential diagnosis of the paravertebral mass when a mass with a hyperintense center and peripheral hypointense rim on T2-weighted images and strong peripheral rim enhancement on contrast-enhanced T1-weighted MR images is seen in postoperative patients. PMID- 17416827 TI - The self-pith. AB - We describe a middle-aged woman who inserted a sewing needle into her spinal cord in an attempt at performing her own acupuncture. Reports of neurologic injury are rare in the literature, despite the widespread use of acupuncture. This is the first case we have identified involving spinal cord injury from self-performed acupuncture. PMID- 17416828 TI - Efficacy and sensitivity of axial scans and different reconstruction methods in the study of the ulcerated carotid plaque using multidetector-row CT angiography: comparison with surgical results. AB - BACKGROUND AND PURPOSE: Carotid plaque ulceration is an important risk factor for stroke, and its diagnosis may be very important to plan a correct therapeutic approach. We hypothesized that axial scans and various reconstruction methods could have different specificity and sensitivity in the study of plaque ulceration. The object of this study was to evaluate their role and diagnostic efficacy in patients with carotid plaque complicated by ulceration through the comparison with surgical results. MATERIALS AND METHODS: From January 2004 to November 2005, 109 patients who underwent a carotid endarterectomy were analyzed using CT angiography for a total of 218 carotid arteries. We assessed every carotid for the presence of ulcerations. For each patient axial image, maximum intensity projection (MIP), multiplanar reconstruction (MPR), shaded surface display (SSD), and volume rendering (VR) reconstructions were obtained. RESULTS: Multidetector row CT angiography (MDCT) found 32 ulcerations; surgical confirmation underlined an overall 93.9% sensitivity (95% confidence interval [CI] 0.858-1.021), and a 98.7% specificity (95% CI, 0.961-1.012). Axial scans and volume rendering images demonstrated the highest sensitivity (90.9% and 87.9%, respectively); SSD, on the contrary, showed the lowest sensitivity: 39.4% (95% CI sensitivity, 0.227-0.561). CONCLUSION: Axial scans plus VR reconstruction techniques offer superior depiction of carotid plaque ulceration compared with MIP, MPR, and SSD. PMID- 17416829 TI - Diffusion tensor imaging-based fractional anisotropy quantification in the corticospinal tract of patients with amyotrophic lateral sclerosis using a probabilistic mixture model. AB - BACKGROUND AND PURPOSE: In amyotrophic lateral sclerosis (ALS), fiber degeneration within the corticospinal tract (CST) can be quantified by diffusion tensor imaging (DTI) as an indirect marker of upper motor neuron involvement. A new method of measuring quantitative DTI parameters using a probabilistic mixture model for fiber tissue and background in the corticospinal tract of patients with ALS is evaluated. MATERIALS AND METHODS: Axial echo-planar imaging (EPI) DTI datasets (6 gradient directions, 10 repetitions) were acquired for 10 patients and 20 healthy control subjects. The diffusion tensor was visualized in a multiplanar viewer using a unique color coding method. Pure fiber tissue inside a region is separated from background and mixture voxels using a probabilistic mixture model. This allows for a reduction of errors as a result of partial volume effects and measurement variability. RESULTS: Fractional anisotropy (FA) was measured within the CST at levels ranging from internal capsule to pons. Mean coefficients of variation of intrarater, scan-rescan, and inter-rater reproducibility were 2.4%, 3.0%, and 5.7%, respectively. Optimal measurement positions along the CST with respect to minimum variability and maximum difference between patients and healthy subjects were identified in the caudal half of the internal capsule. Moreover, a negative correlation between the age corrected FA and the disease duration but not the ALS Severity scale score was found. CONCLUSION: The new software for fiber integrity quantification is suited to assess FA in the corticospinal tract with high reproducibility. Thus, this tool can be useful in future studies for monitoring disease status and potential treatment efficiency. PMID- 17416830 TI - Automated discrimination between very early Alzheimer disease and controls using an easy Z-score imaging system for multicenter brain perfusion single-photon emission tomography. AB - BACKGROUND AND PURPOSE: In Alzheimer disease (AD), a peculiar regional cerebral blood flow (rCBF) abnormality has been reported in the posterior cingulate gyri and precunei, even at a very early stage. We performed a multicenter brain perfusion single-photon emission tomography (SPECT) study to evaluate the discrimination ability of an easy Z-score imaging system (eZIS) with a common normal data base between patients with very early AD at the stage of mild cognitive impairment and age-matched healthy controls. MATERIALS AND METHODS: For a multicenter study, SPECT images of 40 patients with AD and 40 healthy volunteers were acquired from 4 gamma camera systems in 4 different institutions. Systematic differences of SPECT images between gamma cameras were corrected by using conversion maps calculated from the SPECT images of the same brain phantom. Receiver operating characteristic (ROC) analysis was performed to discriminate patients and controls by using a Z-score in the volume of interest (VOI), which had been defined as a region related to AD in subjects other than those in a multicenter study. RESULTS: Bilateral posterior cingulate gyri, precunei, and parietal cortices were defined as a VOI showing rCBF reduction in very early AD. A new indicator of rCBF abnormality in the VOI provided 86% accuracy for distinction of AD and healthy controls in the multicenter study. The area under the ROC curve was 0.934. CONCLUSION: Because an eZIS can use a common normal data base by converting site-specific SPECT data to the core data, the eZIS was useful for automated diagnosis of very early AD in routine studies in multiple institutions. PMID- 17416831 TI - Prediction of cerebral hyperperfusion after carotid endarterectomy using cerebral blood volume measured by perfusion-weighted MR imaging compared with single photon emission CT. AB - BACKGROUND AND PURPOSE: Cerebral hyperperfusion syndrome is a rare but serious complication of carotid revascularization, including carotid endarterectomy (CEA) and carotid stent placement, which can occur in patients with preoperative impairments in cerebral hemodynamics. The purpose of this study was to determine whether preoperative cerebral blood volume (CBV) measured by perfusion-weighted MR imaging (PWI) could identify patients at risk for cerebral hyperperfusion after CEA. MATERIALS AND METHODS: CBV was measured by using PWI before CEA in 70 patients with unilateral internal carotid artery (ICA) stenosis (>or=70%) and without contralateral ICA steno-occlusive disease. Cerebral blood flow (CBF) was also measured by using single-photon emission CT before and immediately after CEA and on the 3rd postoperative day. RESULTS: A significant correlation was observed between preoperative CBV and increases in CBF immediately after CEA (r = 0.785, P < .0001). Whereas hyperperfusion immediately after CEA (CBF increase of >or=100% compared with preoperative values) was observed in 7 of 15 patients (47%) with elevated preoperative CBV, no patients with normal preoperative CBV exhibited post-CEA hyperperfusion. Furthermore, elevated preoperative CBV was the only significant independent predictor of post-CEA hyperperfusion. Finally, hyperperfusion syndrome developed on the 5th postoperative day in 2 of the 7 patients who displayed hyperperfusion immediately after CEA. CONCLUSION: Measurements of preoperative CBV by PWI might help to identify patients at risk for cerebral hyperperfusion after CEA in the absence of contralateral ICA steno occlusive disease. PMID- 17416832 TI - Serial changes in CT cerebral blood volume and flow after 4 hours of middle cerebral occlusion in an animal model of embolic cerebral ischemia. AB - BACKGROUND AND PURPOSE: Neuroimaging techniques have the potential to improve acute stroke treatment by selecting the appropriate patients for thrombolytic therapy. In this study, we examined changes in cerebral blood flow (CBF) and cerebral blood volume (CBV) in an animal model of middle cerebral artery occlusion and used these to identify the parameters that best differentiate between oligemic and infarct regions. MATERIALS AND METHODS: Permanent middle cerebral artery occlusion was performed in 17 New Zealand white rabbits. CT perfusion imaging was performed before (baseline), 10, and 30 minutes after the stroke, and then every 30 minutes up to 3 hours. After a final scan at 4 hours, the brain was removed, cut corresponding to CT sections, and stained with 2,3,5 triphenyltetrazolium chloride (TTC) to identify infarcted tissue. A logistic regression model with the 4-hour post-CBF and -CBV values as independent variables was used to determine the binary tissue outcome variable (oligemia or infarction). RESULTS: Infarcted regions were characterized by a significant decrease (P < .005) in both CBV and CBF, whereas oligemic (CBF < 25 mL . 100 g( 1) . min(-1), not infarcted) regions showed a significant decrease (P < .005) in CBF with maintenance of CBV at or near baseline values. From the perfusion parameters at the 4-hour time point, logistic regression by using CBV*CBF resulted in a sensitivity of 90.6% and a specificity of 93.3% for infarction. CONCLUSION: CBF and CBV values obtained from CT perfusion imaging can be used to distinguish between oligemic and infarct regions. This information could be used to assess the viability of ischemic brain tissue. PMID- 17416833 TI - CT angiography and perfusion CT in cerebral vasospasm after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: We investigated the association of multisection CT angiography (MSCTA) and perfusion CT (PCT) for the characterization of vasospasm secondary to aneurysmal subarachnoid hemorrhage. MATERIALS AND METHODS: Among 27 patients with symptomatic cerebrovascular vasospasm investigated by digital subtraction angiography (DSA), 18 underwent both cerebral PCT and MSCTA. For the remaining 9, only PCT or MSCTA could be performed. MSCTA was compared with DSA for the detection and characterization of vasospasm on 286 intracranial arterial segments. PCT maps were visually reviewed for mean transit time, relative cerebral blood flow, and relative cerebral blood volume abnormalities and were qualitatively compared with the corresponding regional vasospasm detected by DSA. RESULTS: Vasospasm was grouped into 2 categories: mild-moderate and severe. The depiction of vasospasm by MSCTA showed the best sensitivity, specificity, and accuracy at the level of the A2 and M2 arterial segments (100% for each), in contrast to the carotid siphon (45%, 100%, and 85% respectively). The characterization of vasospasm severity by MSCTA showed a sensitivity, specificity, and accuracy of 86.8%, 96.8%, and 95.2%, respectively, for mild moderate vasospasm, and 76.5%, 99.5%, and 97.5%, respectively, for severe vasospasm. The PCT abnormalities were related to severe vasospasm in 9 patients and to mild-to-moderate vasospasm in 2. The sensitivity, specificity, and accuracy of PCT in detecting vasospasm were 90%, 100%, and 92.3%, respectively, for severe vasospasm, and 20%, 100%, and 38.5%, respectively, for mild-moderate vasospasm. CONCLUSION: MSCTA/PCT can assess the location and severity of cerebrovascular vasospasm and its related perfusion abnormalities. It can identify severe vasospasm with risk of delayed ischemia and can thus guide the invasive treatment. PMID- 17416834 TI - Acute Korsakoff-like amnestic syndrome resulting from left thalamic infarction following a right hippocampal hemorrhage. AB - Korsakoff-like amnestic syndromes have been rarely described following structural lesions of the central nervous system. In this report, we describe a case of acute Korsakoff-like syndrome resulting from the combination of a left anteromedian thalamic infarct and a right hippocampal hemorrhage. We also review the literature relevant to the neuropathology and pathophysiology of Korsakoff syndrome and anterograde amnesia. PMID- 17416835 TI - Histogram analysis versus region of interest analysis of dynamic susceptibility contrast perfusion MR imaging data in the grading of cerebral gliomas. AB - BACKGROUND AND PURPOSE: Histogram analysis can be applied to dynamic susceptibility contrast (DSC) perfusion MR imaging datasets and can be as effective as traditional region-of-interest (ROI) measurements of relative cerebral blood volume (rCBV), an operator-dependent method. We compare the routine ROI method with histogram analysis in the grading of glial neoplasms. MATERIALS AND METHODS: Ninety-two patients underwent conventional and DSC MR imaging. Routine rCBV (rCBVmax) measurements were obtained from ROIs of the maximal abnormality within the glioma. Histogram analysis rCBVT was performed with an ROI drawn around the maximal tumor diameter. Spearman rank correlations measured associations among glioma grade, rCBVmax, and histogram measures. Mann Whitney tests compared grade with respect to rCBV and histogram measures. Logistic regression and McNemar test compared the utility of rCBVmax and histogram measures for detecting high grade gliomas. RESULTS: Routine rCBVmax analysis showed significant correlation with grade (r = 0.734, P < .001). Histogram rCBVT metrics showed significant correlation with grade (P < .008); the 3 highest were rCBVT SD, SD50, and mean25 (r = 0.718, 0.684, and 0.683, respectively). Grade could be predicted by rCBVmax (P < .001) as well as rCBV(T) (P < .008). Three rCBVT histogram measures (SD, SD25, and SD50) detected high grade glioma with significantly higher specificity than rCBVmax when the diagnostic tests were constrained to have at least 95% sensitivity. CONCLUSION: rCBVT histogram analysis is as effective as rCBVmax analysis in the correlation with glioma grade. Inexperienced operators may obtain perfusion metrics using histogram analyses that are comparable with those obtained by experienced operators using ROI analysis. PMID- 17416836 TI - Correlation of diffusion tensor and dynamic perfusion MR imaging metrics in normal-appearing corpus callosum: support for primary hypoperfusion in multiple sclerosis. AB - BACKGROUND AND PURPOSE: Hypoperfusion of the normal-appearing white matter in multiple sclerosis (MS) may be related to ischemia or secondary to hypometabolism from wallerian degeneration (WD). This study evaluated whether correlating perfusion and diffusion tensor imaging (DTI) metrics in normal-appearing corpus callosum could provide support for an ischemic mechanism for hypoperfusion. MATERIALS AND METHODS: Fourteen patients with relapsing-remitting MS (RRMS) and 17 control subjects underwent perfusion MR imaging and DTI. Absolute measures of cerebral blood volume (CBV), cerebral blood flow (CBF), and mean transit time (MTT) were calculated. Mean diffusivity (MD) and fractional anisotropy (FA) maps were computed from DTI data. After visual coregistration of perfusion and DTI images, regions of interest were placed in the genu, central body, and splenium of normal-appearing corpus callosum. Pearson product-moment correlation coefficients were calculated using mean DTI and perfusion measures in each region. RESULTS: In the RRMS group, CBF and CBV were significantly correlated with MD in the splenium (r = 0.83 and r = 0.63, respectively; both P < .001) and in the central body (r = 0.86 and r = 0.65, respectively; both P < .001), but not in the genu (r = 0.23 and 0.25, respectively; both P is nonsignificant). No significant correlations were found between MTT and DTI measures or between FA and any perfusion measure in the RRMS group. No significant correlations between diffusion and perfusion metrics were found in control subjects. CONCLUSION: In the normal-appearing corpus callosum of patients with RRMS, decreasing perfusion is correlated with decreasing MD. These findings are more consistent with what would be expected in primary ischemia than in secondary hypoperfusion from WD. PMID- 17416837 TI - Contribution of diffusion tensor MR imaging in detecting cerebral microstructural changes in adults with neurofibromatosis type 1. AB - BACKGROUND AND PURPOSE: After an early progression of signal intensity changes in T2-weighted MR images, also known as "neurofibromatosis bright objects," in patients with neurofibromatosis type 1 (NF-1), there is a tendency toward regression or even disappearance in early adulthood. The purpose of this study was to investigate whether adult patients with NF-1 exhibit generalized microstructural alterations even in normal-appearing brain regions. MATERIALS AND METHODS: Conventional and diffusion tensor MR imaging of the brain was obtained in 10 adult patients with NF-1 and 10 age-matched healthy volunteers. Apparent diffusion coefficient (ADC) and fractional anisotropy (FA) were measured in brain stem, basal ganglia, thalamus, corpus callosum, and frontal and parietooccipital white matter regions. RESULTS: Significantly increased ADC and decreased FA values were found in all regions of interest and in all patients with NF-1, irrespective of their scholastic achievement and subsequent professional performance, compared with control subjects (P < .001). There were no significant correlations with the age (P > .1) or with the lateralization between brain hemispheres (P > .05). CONCLUSION: Diffusion tensor imaging reveals globally elevated FA and decreased ADC values in the mature brains of patients with NF-1, which is most likely a consequence of diffuse and basic alterations in cerebral microstructure that result from the underlying gene mutation. PMID- 17416838 TI - Hyperintense putaminal rim at 3T reflects fewer ferritin deposits in the lateral marginal area of the putamen. AB - BACKGROUND AND PURPOSE: The aim of this study was to clarify the cause of hyperintense putaminal rim (HPR) on the basis of 3T MR imaging-pathologic correlations. MATERIALS AND METHODS: We evaluated brain MR images from 75 subjects 13 to 85 years of age on T2-weighted fast spin-echo (FSE) images at 3T. We also assessed HPR on postmortem T2-weighted FSE images from 4 postmortem cases 1, 12, 63, and 83 years of age. To clarify the cause of HPR, we used 3 staining methods: the Kluver-Barrera method to observe the myelin sheath, the Berlin blue method to observe hemosiderin, and ferritin immunohistochemistry to observe ferritin. The postmortem MR images were compared with the histologic findings in each case. RESULTS: HPR was absent or vague in subjects under 30 years of age but present in subjects in their 30s-60s and again became vague in those subjects older than 70 years of age. The postmortem MR imaging-pathologic correlations revealed that ferritin deposits were slight in the lateral marginal area of the putamen in the 63-year-old subject showing present HPR, but in the 83-year-old subject with no HPR, ferritin deposits were prominent in the lateral marginal area of the putamen as well as in other areas. CONCLUSION: Age-related disproportion in ferritin deposits between the lateral marginal area and the remainder of the putamen causes hypointensity of the latter and the relative hyperintensity of the former, which is depicted as HPR with 3T MR imaging. PMID- 17416839 TI - Partially recanalized chronic dural sinus thrombosis: findings on MR imaging, time-of-flight MR venography, and contrast-enhanced MR venography. AB - BACKGROUND AND PURPOSE: The imaging appearance of chronic, partially recanalized dural sinus thrombosis has been incompletely described. We sought to more fully characterize the imaging findings of this entity on MR imaging, time-of-flight MR venography (TOF-MRV), and elliptic centric-ordered contrast-enhanced MR venography (CE-MRV). MATERIALS AND METHODS: From a data base of patients with cerebral venous thrombosis, 10 patients were identified with imaging and clinical findings consistent with the diagnosis of chronic, partially recanalized, dural sinus thrombosis. All patients had MR imaging of the brain without and with contrast. Nine patients underwent MRV, and 6 had both CE-MRV and TOF-MRV. Thirty four venous segments were thrombosed and were assessed in detail for multiple imaging features. RESULTS: Most thrombosed segments were isointense to gray matter on T1-weighted images (85%), and hyperintense to gray matter on T2 weighted images (97%). Visible serpiginous intrathrombus flow voids were visible in 23 segments (8/10 patients) corresponding with areas of flow signal intensity on TOF-MRV and enhancing channels on contrast MRV. Eighty-four percent of thrombosed segments enhanced equal to or greater than venographically normal venous sinuses. TOF-MRV and CE-MRV were abnormal in all patients, and CE-MRV more completely characterized the thrombosed segments. The imaging appearance did not change in those patients with follow-up imaging (average 13.6 months). CONCLUSION: Chronic, partially recanalized, venous thrombosis has a characteristic appearance on MR and MRV. CE-MRV was abnormal in all cases, despite the intense enhancement of the thrombosed segments. Because of the highly selected nature of the cases reported, further study is required to determine whether these findings are present in all cases of this condition. PMID- 17416840 TI - Utility of navigator-prospective acquisition correction technique (PACE) for reducing motion in brain MR imaging studies. AB - In this retrospective review, we demonstrate the utility of using a commercially available gated navigator sequence (prospective acquisition correction technique [PACE]) to reduce rhythmic breathing motion on brain MR images. For purposes of this report, we studied 2 sedated patients who had marked head rocking, one due to deep breathing and snoring and the other due to ventilator support. Motion degraded the routine images, which, despite a short acquisition time, were nondiagnostic. After application of PACE, a technique commonly used in abdominal studies, the brain images in both patients were judged to be of acceptable diagnostic quality. The use of PACE to diminish head motion is now routine at our institution when sedated or ventilated patients degrade images with involuntary head rocking. PMID- 17416848 TI - The quest for ovarian cancer's Holy Grail: can CA-125 still be the chalice of early detection? PMID- 17416849 TI - On the TRAIL toward death receptor-based cancer therapeutics. PMID- 17416850 TI - Cutting back on computed tomography scan frequency in curative oncology: get the picture. PMID- 17416851 TI - Randomized trial of two or five computed tomography scans in the surveillance of patients with stage I nonseminomatous germ cell tumors of the testis: Medical Research Council Trial TE08, ISRCTN56475197--the National Cancer Research Institute Testis Cancer Clinical Studies Group. AB - PURPOSE: Surveillance is a standard management approach for stage I nonseminomatous germ cell tumors (NSGCT). A randomized trial of two versus five computed tomography (CT) scans was performed to determine whether the number of scans influenced the proportion of patients relapsing with intermediate- or poor prognosis disease at relapse. METHODS: Patients with clinical stage I NSGCT opting for surveillance were randomly assigned to chest and abdominal CT scans at either 3 and 12 or 3, 6, 9, 12, and 24 months, with all other investigations identical in the two arms. Three of five patients were allocated to the two-scan schedule. Four hundred patients were required. RESULTS: Two hundred forty-seven patients were allocated to a two-scan and 167 to five-scan policy. With a median follow-up of 40 months, 37 relapses (15%) have occurred in the two-scan arm and 33 (20%) in the five-scan arm. No patients had poor prognosis at relapse, but two (0.8%) of those relapsing in the two-scan arm had intermediate prognosis compared with 1 (0.6%) in the five-scan arm, a difference of 0.2% (90% CI, -1.2% to 1.6%). No deaths have been reported. CONCLUSION: This study can rule out with 95% probability an increase in the proportion of patients relapsing with intermediate or poor-prognosis disease of more than 1.6% if they have two rather than five CT scans as part of their surveillance protocol. CT scans at 3 and 12 months after orchidectomy should be considered a reasonable option in low-risk patients. PMID- 17416852 TI - Multi-institutional validation of a new renal cancer-specific survival nomogram. AB - PURPOSE: We tested the hypothesis that the prediction of renal cancer-specific survival can be improved if traditional predictor variables are used within a prognostic nomogram. PATIENTS AND METHODS: Two cohorts of patients treated with either radical or partial nephrectomy for renal cortical tumors were used: one (n = 2,530) for nomogram development and for internal validation (200 bootstrap resamples), and a second (n = 1,422) for external validation. Cox proportional hazards regression analyses modeled the 2002 TNM stages, tumor size, Fuhrman grade, histologic subtype, local symptoms, age, and sex. The accuracy of the nomogram was compared with an established staging scheme. RESULTS: Cancer specific mortality was observed in 598 (23.6%) patients, whereas 200 (7.9%) died as a result of other causes. Follow-up ranged from 0.1 to 286 months (median, 38.8 months). External validation of the nomogram at 1, 2, 5, and 10 years after nephrectomy revealed predictive accuracy of 87.8%, 89.2%, 86.7%, and 88.8%, respectively. Conversely, the alternative staging scheme predicting at 2 and 5 years was less accurate, as evidenced by 86.1% (P = .006) and 83.9% (P = .02) estimates. CONCLUSION: The new nomogram is more contemporary, provides predictions that reach further in time and, compared with its alternative, which predicts at 2 and 5 years, generates 3.1% and 2.8% more accurate predictions, respectively. PMID- 17416853 TI - Meta-analysis of BRCA1 and BRCA2 penetrance. AB - PURPOSE: Genetic counseling is now routinely offered to individuals at high risk of carrying a BRCA1 or BRCA2 mutation. Risk prediction provided by the counselor requires reliable estimates of the mutation penetrance. Such penetrance has been investigated by studies worldwide. The reported estimates vary. To facilitate clinical management and counseling of the at-risk population, we address this issue through a meta-analysis. METHODS: We conducted a literature search on PubMed and selected studies that had nonoverlapping patient data, contained genotyping information, used statistical methods that account for the ascertainment, and reported risks in a useable format. We subsequently combined the published estimates using the DerSimonian and Laird random effects modeling approach. RESULTS: Ten studies were eligible under the selection criteria. Between-study heterogeneity was observed. Study population, mutation type, design, and estimation methods did not seem to be systematic sources of heterogeneity. Meta-analytic mean cumulative cancer risks for mutation carriers at age 70 years were as follows: breast cancer risk of 57% (95% CI, 47% to 66%) for BRCA1 and 49% (95% CI, 40% to 57%) for BRCA2 mutation carriers; and ovarian cancer risk of 40% (95% CI, 35% to 46%) for BRCA1 and 18% (95% CI, 13% to 23%) for BRCA2 mutation carriers. We also report the prospective risks of developing cancer for currently asymptomatic carriers. CONCLUSION: This article provides a set of risk estimates for BRCA1 and BRCA2 mutation carriers that can be used by counselors and clinicians who are interested in advising patients based on a comprehensive set of studies rather than one specific study. PMID- 17416854 TI - Understanding of prognosis among parents of children with cancer: parental optimism and the parent-physician interaction. AB - PURPOSE: Patients often overestimate their chances of surviving cancer. Factors that contribute to accurate understanding of prognosis are not known. We assessed understanding of likelihood of cure and functional outcome among parents of children with cancer and sought to identify factors that place parents at risk for overly optimistic beliefs about prognosis. PATIENTS AND METHODS: We conducted a cross-sectional survey of 194 parents of children with cancer (response rate, 70%) who were treated at the Dana-Farber Cancer Institute and Children's Hospital in Boston, MA, and the children's physicians. Parent and physician expectations for likelihood of cure and functional outcome were compared. In 152 accurate or optimistic parents, we determined factors associated with accurate understanding of likelihood of cure compared with optimism. RESULTS: The majority of parents (61%) were more optimistic than physicians about the likelihood of cure. Parents' beliefs about other outcomes of cancer treatment were similar (quality-of-life impairment, P = .70) or more pessimistic (physical impairment, P = .01; intellectual impairment, P = .01) than physicians' beliefs. Parents and physicians were more likely to agree about chances of cure when physicians had confidence in knowledge of prognosis (odds ratio [OR] = 2.55, P = .004) and allowed parents to take their preferred decision-making role (OR = 1.89, P = .019). CONCLUSION: Parents of children with cancer are overly optimistic about chances of cure but not about other outcomes of cancer therapy. Parents tend to be overly optimistic about cure when physicians have little confidence and when the decision-making process does not meet parents' preferences. These findings suggest that physicians are partly responsible for parents' unrealistic expectations about cure. PMID- 17416855 TI - Melanoma in children and teenagers: an analysis of patients from the National Cancer Data Base. AB - PURPOSE: This study examines the demographics, presentation, and outcomes of children and teenagers with melanoma using a US hospital-based oncology database. PATIENTS AND METHODS: Data from the National Cancer Data Base from 1985 through 2003 were examined for demographics, presentation, and survival of patients aged 1 to 19 years, as well as a comparison group of patients aged 20 to 24 years. Two sided linear and Pearson chi2 tests were calculated to examine associations. Proportions were compared using two-sided z tests. Five-year overall observed survival was evaluated using the Kaplan-Meier method and the log-rank test. Cox proportional hazards regression was used to estimate risk of mortality. RESULTS: Of 3,158 patients aged 1 to 19 years, 96.3% had cutaneous melanoma, 3.0% had ocular melanoma, and 0.7% had an unknown primary tumor. Cutaneous melanoma in patients aged 1 to 19 years was more common in girls (55.5%) and patients older than 10 years (90.5%). The demographics and presentation of cutaneous melanoma were age related; younger children were significantly more likely to be nonwhite and male and more likely to present with a head and neck primary tumors and with regional or distant metastases (linear chi2, P < .001 for sex, race, and extent of disease). Poorer survival was associated with higher stage and younger age. In contrast to patients aged 20 to 24 years, survival was not related to thickness in patients aged 1 to 19 years with localized invasive melanoma. CONCLUSION: Melanoma in children and teenagers differs from melanoma in young adults in demographics, presentation, and survival. Further investigation is warranted to elucidate possible biologic correlates of the unique aspects of melanoma in children and teenagers. PMID- 17416856 TI - Identification of angiogenesis/metastases genes predicting chemoradiotherapy response in patients with laryngopharyngeal carcinoma. AB - PURPOSE: To identify genes related to angiogenesis/metastasis that predict locoregional failure in patients with laryngopharyngeal cancer (LPC) undergoing chemoradiotherapy (CRT) treatment. METHODS: Tumor tissue was collected and snap frozen from 35 sequential patients with histologically confirmed LPC being treated with CRT. Gene expression analysis was performed using a novel cDNA array consisting of 277 genes functionally associated with angiogenesis (n = 152) and/or metastasis (n = 125). Locoregional response was correlated to the gene expression profiles to identify genes associated with outcome. These genes were internally validated by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and validated externally by immunohistochemistry analysis on an independent set of patients. RESULTS: Locoregional failure occurred in nine of 35 patients. Seventeen genes from the cDNA microarray correlated with locoregional failure (two-sample t test, P < .05). Seven genes were chosen for additional analysis based on the availability of antibodies for immunohistochemistry. Of these seven genes, real-time RT-PCR validated four genes: MDM2, VCAM-1, erbB2, and H-ras (Wilcoxon rank sum test, P = .008, .02, .04, and .04, respectively). External validation by immunohistochemistry confirmed MDM2 and erbB2 as being predictive of locoregional response. Controlling for stage of disease, positivity for MDM2 or erbB2 was an independent negative predictor of locoregional disease free survival. CONCLUSION: Genomic screening by cDNA microarray and validation internally by real-time RT-PCR and externally by immunohistochemistry have identified two genes (MDM2 and erbB2) as predictors of locoregional failure in LPC patients treated with CRT. The role of these genes in treatment selection and the functional basis for their activity in CRT response merit additional consideration. PMID- 17416857 TI - Individual patient data meta-analysis of docetaxel administered once every 3 weeks compared with once every week second-line treatment of advanced non-small cell lung cancer. AB - PURPOSE: Although several randomized trials have been performed comparing weekly docetaxel (wD) with standard docetaxel once every 3 weeks (3wD) as second-line treatment of advanced non-small-cell lung cancer (NSCLC), no single trial had sufficient power to detect clinically relevant differences in survival. METHODS: We performed a meta-analysis based on individual patient data from all identified randomized trials comparing wD with 3wD as second-line treatment of advanced NSCLC. Baseline characteristics, treatment assigned, and outcome data were collected for each patient. The primary end point was overall survival. All statistical analyses were stratified by trial. RESULTS: Five eligible trials were identified for a total of 865 patients: 433 patients had been assigned to 3wD, and 432 patients had been assigned to wD. Median age was 62 years (range, 26 to 80 years). Performance status was 0 in 23%, 1 in 58%, and 2 in 16% of patients; 91% of the patients had received previous platinum, and 14% had received previous paclitaxel. With 733 deaths recorded (85%), median survival was 27.4 weeks for patients treated with 3wD, and 26.1 weeks for patients treated with wD (P = .24, log-rank test). There was no significant heterogeneity among the five trials. No relevant differential effect was detected in subgroup analyses. Significantly less severe and febrile neutropenia was reported with wD (P < .00001 for both), whereas no significant differences were observed for anemia, thrombocytopenia, and nonhematologic toxicity. CONCLUSION: wD shows similar efficacy compared with 3wD, and represents an alternative for second-line treatment of advanced NSCLC. PMID- 17416858 TI - Serum CA-125 in relation to adnexal dysplasia and cancer in women at hereditary high risk of ovarian cancer. AB - PURPOSE: Serum CA-125 level is commonly used as indicator for ovarian cancer recurrence. However, its value for the prediction of neoplastic lesions is unknown. The aim of this study was to investigate whether CA-125 concentrations are indicative of adnexal dysplasia and cancer in women at hereditary high risk of ovarian/tubal cancer. PATIENTS AND METHODS: CA-125 was obtained from 424 women at hereditary high risk of ovarian/tubal cancer attending the VU University Medical Center (Amsterdam, the Netherlands) between 1993 and 2005. Serum samples obtained at the second-to-last (n = 64) and last (n = 98) visit before surgery were tested in women who underwent adnexal surgery for diagnostic (n = 9) or prophylactic (n = 89) reasons. Serum samples obtained from 370 age-matched healthy women were used as controls. RESULTS: Both the absolute value (P < .0001) and the serial change (P < .0001) of CA-125 were predictive for ovarian cancer (n = 8). For adnexal dysplasia (n = 23), the absolute value of CA-125 (P = .003) was predictive, but the serial change in CA-125 was not (P = .32). The odds ratio for adnexal dysplasia versus nondysplasia in the highest tertile (CA-125 levels 14 U/mL) compared with the lowest tertile (CA-125 < 10 U/mL) was 6 (95% CI, 1.32 to 36.66). CONCLUSION: In patients at hereditary high risk for adnexal cancer, both the absolute value of serum CA-125 and the change in serial CA-125 are predictors for ovarian cancer. Remarkably, the absolute value of CA-125 is also predictive for adnexal dysplasia. CA-125 values should, therefore, be taken into account in the decision toward prophylactic bilateral salpingo-oophorectomy. PMID- 17416859 TI - Phase I pharmacokinetic and biologic correlative study of mapatumumab, a fully human monoclonal antibody with agonist activity to tumor necrosis factor-related apoptosis-inducing ligand receptor-1. AB - PURPOSE: To assess the safety, pharmacokinetics, and preliminary evidence of antitumor activity of mapatumumab (HGS-ETR1, TRM-1), a fully human agonist monoclonal antibody directed to the tumor necrosis factor-related apoptosis inducing ligand receptor-1 (TRAIL-R1). PATIENTS AND METHODS: Patients with advanced solid malignancies were treated with escalating doses of mapatumumab intravenously (IV) administered over 30 to 120 minutes, initially as a single dose and then repetitively. Plasma mapatumumab concentrations were measured and serum was assayed to detect human antimapatumumab antibody formation. Archival tumor specimens were collected to detect the presence of TRAIL-R1 by immunohistochemistry. RESULTS: Forty-nine patients received 158 courses at doses ranging from 0.01 to 10 mg/kg IV. Initially, patients received mapatumumab as a single dose, then every 28 days repetitively, and then 10 mg/kg every 14 days. Mild (grade 1 or 2) fatigue, fever, and myalgia were the most frequently reported nonhematologic adverse events related to mapatumumab, whereas hematologic toxicity was not clinically significant. The mean (+/- standard deviation) clearance and terminal elimination half-life values for mapatumumab at 10 mg/kg every 14 days were 3.7 mL/d/kg (+/- 1.5 mL/d/kg) and 18.8 days (+/- 10.1 days), respectively. TRAIL-R1 was documented in 68% of patients' tumors assayed. Nineteen patients had stable disease, with two lasting 9 months. CONCLUSION: Mapatumumab can be administered safely and feasibly at 10 mg/kg IV every 14 days. The absence of severe toxicities and the attainment of plasma mapatumumab concentrations that are active in preclinical models warrant further disease directed studies of this agent alone and in combination with chemotherapy in a broad array of tumors. PMID- 17416860 TI - Late effects of cancer and hematopoietic stem-cell transplantation on spouses or partners compared with survivors and survivor-matched controls. AB - PURPOSE: Little is known about the long-term effects of cancer and hematopoietic stem-cell transplantation (HCT) on spouses or partners. The purpose of this study was to examine the health-related quality of life and post-traumatic growth (PTG) of spouses/partners compared with survivors and controls and to identify factors associated with those outcomes. PATIENTS AND METHODS: HCT survivor/partner pairs (n = 177), coupled continuously since HCT, were drawn from 40 North American transplantation centers. Married peer-nominated acquaintances (of survivors) served as controls (n = 133). Outcomes were measured a mean of 6.7 years after HCT (range, 1.9 to 19.4 years). RESULTS: As expected, self-reported partner physical health was similar to controls and better than survivors (P < .001). However, partners reported more fatigue and cognitive dysfunction than controls (P < .001 for both), although less than survivors. Partners and survivors reported more depressive symptoms and sleep and sexual problems than controls (P < .001, P < .01, and P < .01, respectively). Odds of partner depression were nearly 3.5 times that of controls (P < .002). Depressed partners were less likely than depressed survivors to receive mental health treatment (P < .04). Partners reported less social support (P < .001), dyadic satisfaction (P < .05), and spiritual well-being (P < .05) and more loneliness (P < .05) than both survivors and controls. In contrast to survivors, partners reported little PTG (P < .001). Factors associated with partner outcomes included partner health problems, coping, female sex, social constraint, survivor depression, optimism, multiple life changes, and social support. CONCLUSION: Spouses/partners experience similar emotional and greater social long-term costs of cancer and HCT than survivors without the potential compensatory benefits of PTG. Some of the factors associated with partner outcomes are amenable to intervention. PMID- 17416861 TI - Does one person provide it all? Primary support and advanced care planning for women with cancer. AB - PURPOSE: We challenged a common clinical assumption that female cancer patients designate the same individual who provides them with primary support in advanced care planning roles. PATIENTS AND METHODS: During 6 months, we conducted a survey of female cancer patients seen in a women's oncology program. Women were asked to name their health care proxy (HCP), emergency contact (EC), and primary support, and to define their own meaning of primary support. The person identified by chart review (C-ID) as that patient's primary contact was also abstracted. The frequency that the same person served all three roles was determined. Concordance between the C-ID and surveyed responses was calculated. RESULTS: Two hundred fifteen (98%) of 219 women agreed to participate. The median age was 58 years (range, 29 to 85 years). Women were surveyed a median of 14.3 months since diagnosis (range, 0.3 to 214 months). Although 80.4% of women surveyed named a first-degree relative to fill all three roles, only 56.7% of respondents identified the same individual for each of these roles. For those in relationships, 54% named their partner in all three roles. The majority of women characterized support as emotional (46.7%) or as emotional and structural (22.6%). Concordance was strongest between the C-ID and both EC and HCP. CONCLUSION: More than 40% of women with cancer in our sample did not name the same person for support and for advanced care planning roles. We also discovered a lack of uniformity in definitions of primary support. The implications of these findings on decision making and outcomes in women with cancer require additional study. PMID- 17416862 TI - PancPRO: risk assessment for individuals with a family history of pancreatic cancer. AB - PURPOSE: The rapid fatality of pancreatic cancer is, in large part, the result of an advanced stage of diagnosis for the majority of patients. Identification of individuals at high risk of developing pancreatic cancer is a first step towards the early detection of this disease. Individuals who may harbor a major pancreatic cancer susceptibility gene are one such high-risk group. The goal of this study was to develop and validate PancPRO, a Mendelian model for pancreatic cancer risk prediction in individuals with familial pancreatic cancer, to identify high-risk individuals. METHODS: PancPRO was built by extending the Bayesian modeling framework developed for BRCAPRO, trained using published data, and validated using independent prospective data on 961 families enrolled onto the National Familial Pancreas Tumor Registry, including 26 individuals who developed incident pancreatic cancer during follow-up. RESULTS: We developed a risk prediction model, PancPRO, and free software for the estimation of pancreatic cancer susceptibility gene carrier probabilities and absolute pancreatic cancer risk. Model validation demonstrated an observed to predicted pancreatic cancer ratio of 0.83 (95% CI, 0.52 to 1.20) and high discriminatory ability, with an area under the receiver operating characteristic curve of 0.75 (95% CI, 0.68 to 0.81) for PancPRO. CONCLUSION: PancPRO is the first risk prediction model for pancreatic cancer. When we validated our model using the largest registry of familial pancreatic cancer, our model provided accurate risk assessment. Our findings highlight the importance of detailed family history for clinical cancer risk assessment and demonstrate that accurate genetic risk assessment is possible even when the causative genes are not known. PMID- 17416863 TI - Palliative radiotherapy trials for bone metastases: a systematic review. AB - PURPOSE: The objective is to update previous meta-analyses with a systematic review of randomized palliative radiotherapy (RT) trials comparing single fractions (SFs) versus multiple fractions (MFs). METHODS: The analysis includes all published reports from randomized trials comparing SF or MF schedules for the treatment of painful bone metastases with localized RT. A systematic review was performed using the random-effects model with Review Manager version 4.1 (Cochrane Collaboration, Oxford, UK). The odds ratio and 95% CI were calculated for each trial and presented in a forest plot. RESULTS: A total of 16 randomized trials from 1986 onward were identified. For intention-to-treat patients, the overall response (OR) rates for pain were similar for SF at 1,468 (58%) of 2,513 patients and MF RT at 1,466 (59%) of 2,487 patients. The complete response (CR) rates for pain were 23% (545 of 2,375 patients) for SF and 24% (558 of 2,351 patients) for MF RT. No significant differences were found in response rates. Trends showing an increased risk for SF RT arm patients in terms of pathological fractures and spinal cord compressions were observed, but neither were statistically significant (P = .75 and P = .13, respectively). The likelihood of re-treatment was 2.5-fold higher (95% CI, 1.76 to 3.56) in SF RT arm patients (P < .00001). Repeated analysis of these end points, excluding dropout patients, did not alter the conclusions. Generally, no significant differences with respect to acute toxicities were observed between the arms. CONCLUSION: No significant differences in the arms were observed for overall and CR rates in both intention to-treat and assessable patients. However, a significantly higher re-treatment rate with SFs was evident. PMID- 17416864 TI - Hodgkin's lymphoma involving the CNS. PMID- 17416865 TI - Trastuzumab-associated peripheral vascular toxicity. PMID- 17416866 TI - Desmoplastic small round-cell tumor of the pancreas. PMID- 17416867 TI - Improving informed consent in clinical trials: successful piloting of a decision aid. PMID- 17416868 TI - Single-agent chemotherapy for the treatment of elderly patients with advanced non small-cell lung cancer: what is the best drug? PMID- 17416869 TI - Do erythropoietin receptors on cancer cells explain unexpected clinical findings? PMID- 17416870 TI - Hyperthermic isolated limb perfusion with tumor necrosis factor is a useful therapy for advanced melanoma of the limbs. PMID- 17416871 TI - p73 and p53 pathway in human breast cancers. PMID- 17416872 TI - Performance of biopsies in clinical research. PMID- 17416873 TI - Changes in bone mineral density after adjuvant aromatase inhibitors and fracture risk in breast cancer patients. PMID- 17416874 TI - Learning implicitly: effects of task and severity after stroke. PMID- 17416875 TI - Do first years really last a lifetime? AB - Multiple early childhood intervention programs have been introduced to provide at risk children with a better start in life. This is due to the recognition that early childhood experiences, both positive and negative, can affect the physical, mental, behavioral, and economic well-being of the child. A few of these programs have evaluated long-term outcomes and have demonstrated that the effects of interventions may still be apparent years after the intervention. Pediatricians need to become more aware of the information available about the impact of early childhood events and become more active in promoting proven best intervention practices at the local, state, and national level. PMID- 17416876 TI - Hemangiomas: an overview. PMID- 17416877 TI - Management of child with fever after international travel. PMID- 17416878 TI - Studying injury prevention: practices, problems, and pitfalls in implementation. AB - This prospective, randomized, controlled trial was conducted to determine feasibility and effectiveness of a chronic care model approach to injury prevention compared with standard anticipatory guidance. Enrolled caregivers of children aged 0 to 5 years received focused counseling from a physician and health assistant, educational handouts, phone follow-up, and access to free safety devices and automobile restraint evaluations. Only 35.1% of eligible parents participated. Home visits were completed at 6 months to observe safety practices. Injuries were gleaned from parent report and medical record review. Safety practices were evaluated in 27 households. Chart review showed no significant difference in the number of medically attended injuries between groups (P = 0.6). The impact of the chronic care model on injury prevention in primary care could not be determined with certainty. Evaluating effectiveness of injury prevention strategies on actual safety practices with direct observation is challenging. PMID- 17416879 TI - Improvement of cardiac auscultation skills in pediatric residents with training. AB - Cardiac murmurs, most of which are harmless, are present in more than 50% of children. Good auscultation skills are required to prevent unnecessary referrals. The auscultation skills of a group of 21 pediatric residents were assessed. Based on their identification of key features such as S1, S2, timing, grade, location, quality of the murmur, and any extra sounds, residents were asked to make a clinical diagnosis. The overall diagnostic accuracy for all residents was 30% and improved with years of training. The average score for correctly identified key features was 46% with no significant improvement with year of training. Residents who had completed a cardiology rotation scored better than the others (41% vs 18% for correct diagnosis). Clinical auscultation skills of pediatric residents in our institution showed significant improvement with training, especially in the ability to recognize a harmless heart murmur. PMID- 17416880 TI - Throat culture is necessary after negative rapid antigen detection tests. AB - This study was conducted to determine if culture confirmation is needed for a negative rapid antigen detection test. Data on 18,509 tests done in patients younger than 18 years old were reviewed. Of the 14,167 (76.5%) that were negative, 968 (6.8%) were associated with positive cultures. No significant seasonal variation was noted. Significant differences were found between hospital and pediatric practices in the percentage of patients with a negative rapid antigen detection test who actually had group A beta-hemolytic streptococcus (3.5% to 9.8%). This study supports the recommendation of culture confirmation of a negative rapid antigen detection test and validation of results within an individual practice if confirmatory cultures are not being performed. This study showed a high false-negative rate of the negative rapid antigen detection test and variation among hospital and pediatric practices for rates of positive culture after a negative rapid antigen detection test. PMID- 17416881 TI - Survival times and complications of catheters used for outpatient parenteral antibiotic therapy in children. AB - Outpatient parenteral antibiotic therapy is routinely used in pediatrics, but few data are available on catheter-associated complications and survival times. Catheter-associated complications, defined as mechanical or nonmechanical, and survival times in peripherally inserted central catheters and central venous catheters used for outpatient parenteral antibiotic therapy in childhood were compared. The life test procedure was performed to determine survival time. Cox proportional hazards model was used to compare the independent effect of variables such as age and gender on catheter survival. There were 104 peripheral and 130 central venous catheters, of which 28 peripheral and 19 central catheters had mechanical complications, and 13 peripheral and 17 central catheters had nonmechanical complications. Peripheral catheters are more likely to develop mechanical complications and have a shorter survival time than central venous catheters. For outpatient parenteral antibiotic therapy longer than 6 weeks, central venous catheters appear to be a better choice. PMID- 17416882 TI - Self-efficacy for smoking cessation counseling parents in primary care: an office based intervention for pediatricians and family physicians. AB - Few pediatricians or family physicians routinely counsel parental smokers to quit smoking. Poor self-efficacy in smoking cessation counseling skills may be one barrier to counseling. Analysis of self-efficacy scores of physicians participating in the Clean Air for Healthy Children program demonstrates that pediatricians had higher self-efficacy scores for explaining the health risks of environmental tobacco smoke on children (P < .05); family physicians had higher self-efficacy scores for smoking cessation counseling knowledge (P < .05). Posttraining, both pediatricians and family physicians who participated in an office-based smoking cessation counseling program had significantly higher scores in all 4 self-efficacy domains (P < .01). PMID- 17416883 TI - Severe human metapneumovirus infection in hospitalized children. AB - Human metapneumovirus is a recently discovered pathogen that causes upper and lower respiratory tract disease in children. This study describes the course of illness in hospitalized children with this infection. During a 6-month period, 11 children were diagnosed with human metapneumovirus infection by reverse transcription-polymerase chain reaction. Oxygen supplementation was required for 82% of patients. Severe disease developed in 45%, and mechanical ventilation was required. An apparent life-threatening event was the indication for hospitalization of 27% of patients infected with human metapneumovirus. Children with underlying asthma or neuromuscular disease had a prolonged hospitalization. PMID- 17416884 TI - Spontaneous spinal epidural hematoma after seizure: a case report. PMID- 17416885 TI - An interesting case of primary autoimmune neutropenia. PMID- 17416886 TI - Pigmented villonodular synovitis causing painless chronic knee swelling in an adolescent. AB - It is uncommon for an adolescent to present with a painless chronic knee effusion without a history of trauma. Acute knee effusions are usually caused by internal derangement, patellar dislocations, or fractures. The differential diagnosis of a chronic knee effusion must include the more uncommon diagnoses of juvenile rheumatoid arthritis, septic arthritis, reactive arthritis, synovial osteochondromatosis, synovial hemangioma, synovial sarcoma, or pigmented villonodular synovitis. Radiographic imaging, plain radiographs, and magnetic resonance imaging in addition to a biopsy specimen are most helpful in differentiating these diagnoses. Pigmented villonodular synovitis is an uncommon diagnosis that is rarely seen in an adolescent. Treatment includes synovectomy via arthroscopy or open arthrotomy. PMID- 17416887 TI - Chronic cocaine exposure in a toddler revealed by hair test. PMID- 17416888 TI - Digoxin toxicity. PMID- 17416889 TI - Neck pain. PMID- 17416890 TI - Dysphonia. PMID- 17416891 TI - Postpartum depression. PMID- 17416892 TI - Role of SP65 in assembly of the Dictyostelium discoideum spore coat. AB - Like the cyst walls of other protists, the spore coat of Dictyostelium discoideum is formed de novo to protect the enclosed dormant cell from stress. Spore coat assembly is initiated by exocytosis of protein and polysaccharide precursors at the cell surface, followed by the infusion of nascent cellulose fibrils, resulting in an asymmetrical trilaminar sandwich with cellulose filling the middle layer. A molecular complex consisting of cellulose and two proteins, SP85 and SP65, is associated with the inner and middle layers and is required for proper organization of distinct proteins in the outer layer. Here we show that, unlike SP85 and other protein precursors, which are stored in prespore vesicles, SP65 is, like cellulose, synthesized just in time. By tagging the SP65 locus with green fluorescent protein, we find that SP65 is delivered to the cell surface via largely distinct vesicles, suggesting that separate delivery of components of the cellulose-SP85-SP65 complex regulates its formation at the cell surface. In support of previous in vivo studies, recombinant SP65 and SP85 are shown to interact directly. In addition, truncation of SP65 causes a defect of the outer layer permeability barrier as seen previously for SP85 mutants. These observations suggest that assembly of the cellulose-SP85-SP65 triad at the cell surface is biosynthetically regulated both temporally and spatially and that the complex contributes an essential function to outer layer architecture and function. PMID- 17416893 TI - Further characterization of the signaling proteolysis step in the Aspergillus nidulans pH signal transduction pathway. AB - The Aspergillus nidulans pH-responsive transcription factor PacC is modulated by limited, two-step proteolysis. The first, pH-regulated cleavage occurs in the 24 residue highly conserved "signaling protease box" in response to the alkaline pH signal. This is transduced by the Pal signaling pathway, containing the predicted calpain-like cysteine protease and likely signaling protease, PalB. In this work, we carried out classical mutational analysis of the putative signaling protease PalB, and we describe 9 missense and 18 truncating loss-of-function (including null) mutations. Mutations in the region of and affecting directly the predicted catalytic cysteine strongly support the deduction that PalB is a cysteine protease. Truncating and missense mutations affecting the C terminus highlight the importance of this region. Analysis of three-hemagglutinin-tagged PalB in Western blots demonstrates that PalB levels are independent of pH and Pal signal transduction. We have followed the processing of MYC(3)-tagged PacC in Western blots. We show unequivocally that PalB is essential for signaling proteolysis and is definitely not the processing protease. In addition, we have replaced 15 residues of the signaling protease box of MYC(3)-tagged PacC (pacC900) with alanine. The majority of these substitutions are silent. Leu481Ala, Tyr493Ala, and Gln499Ala result in delayed PacC processing in response to shifting from acidic to alkaline medium, as determined by Western blot analysis. Leu498Ala reduces function much more markedly, as determined by plate tests and processing recalcitrance. Excepting Leu498, this demonstrates that PacC signaling proteolysis is largely independent of sequence in the cleavage region. PMID- 17416894 TI - Chlamydomonas reinhardtii CNX1E reconstitutes molybdenum cofactor biosynthesis in Escherichia coli mutants. AB - We have isolated and characterized the Chlamydomonas reinhardtii genes for molybdenum cofactor biosynthesis, namely, CNX1G and CNX1E, and expressed them and their chimeric fusions in Chlamydomonas and Escherichia coli. In all cases, the wild-type phenotype was restored in individual mutants as well as in a CNX1G CNX1E double mutant. Therefore, CrCNX1E is the first eukaryotic protein able to complement an E. coli moeA mutant. PMID- 17416895 TI - Barrier activity in Candida albicans mediates pheromone degradation and promotes mating. AB - Mating in Candida albicans and Saccharomyces cerevisiae is regulated by the secretion of peptide pheromones that initiate the mating process. An important regulator of pheromone activity in S. cerevisiae is barrier activity, involving an extracellular aspartyl protease encoded by the BAR1 gene that degrades the alpha pheromone. We have characterized an equivalent barrier activity in C. albicans and demonstrate that the loss of C. albicans BAR1 activity results in opaque a cells exhibiting hypersensitivity to alpha pheromone. Hypersensitivity to pheromone is clearly seen in halo assays; in response to alpha pheromone, a lawn of C. albicans Deltabar1 mutant cells produces a marked zone in which cell growth is inhibited, whereas wild-type strains fail to show halo formation. C. albicans mutants lacking BAR1 also exhibit a striking mating defect in a cells, but not in alpha cells, due to overstimulation of the response to alpha pheromone. The block to mating occurs prior to cell fusion, as very few mating zygotes were observed in mixes of Deltabar1 a and alpha cells. Finally, in a barrier assay using a highly pheromone-sensitive strain, we were able to demonstrate that barrier activity in C. albicans is dependent on Bar1p. These studies reveal that a barrier activity to alpha pheromone exists in C. albicans and that the activity is analogous to that caused by Bar1p in S. cerevisiae. PMID- 17416896 TI - Involvement of a Magnaporthe grisea serine/threonine kinase gene, MgATG1, in appressorium turgor and pathogenesis. AB - We isolated an MgATG1 gene encoding a serine/threonine protein kinase from the rice blast fungus Magnaporthe grisea. In the DeltaMgatg1 mutant, in which the MgATG1 gene had been deleted, autophagy was blocked; the mutant also showed fewer lipid droplets in its conidia, lower turgor pressure of the appressorium, and such defects in morphogenesis as delayed initiation and slower germination of conidia. As a result of lower turgor pressure of the appressorium, the DeltaMgatg1 mutant lost its ability to penetrate and infect the two host plants, namely, rice and barley. However, normal values of the parameters and infective abilities were restored on reintroducing an intact copy of the MgATG1 gene into the mutant. Autophagy is thus necessary for turnover of organic matter during the formation of conidia and appressoria and for normal development and pathogenicity in M. grisea. PMID- 17416897 TI - Genomic and biochemical analysis of lipid biosynthesis in the unicellular rhodophyte Cyanidioschyzon merolae: lack of a plastidic desaturation pathway results in the coupled pathway of galactolipid synthesis. AB - The acyl lipids making up the plastid membranes in plants and algae are highly enriched in polyunsaturated fatty acids and are synthesized by two distinct pathways, known as the prokaryotic and eukaryotic pathways, which are located within the plastids and the endoplasmic reticulum, respectively. Here we report the results of biochemical as well as genomic analyses of lipids and fatty acids in the unicellular rhodophyte Cyanidioschyzon merolae. All of the glycerolipids usually found in photosynthetic algae were found, such as mono- and digalactosyl diacylglycerol, sulfolipid, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. However, the fatty acid composition was extremely simple. Only palmitic, stearic, oleic, and linoleic acids were found as major acids. In addition, 3-trans-hexadecanoic acid was found as a very minor component in phosphatidylglycerol. Unlike the case for most other photosynthetic eukaryotes, polyenoic fatty acids having three or more double bonds were not detected. These results suggest that polyunsaturated fatty acids are not necessary for photosynthesis in eukaryotes. Genomic analysis suggested that C. merolae lacks acyl lipid desaturases of cyanobacterial origin as well as stearoyl acyl carrier protein desaturase, both of which are major desaturases in plants and green algae. The results of labeling experiments with radioactive acetate showed that the desaturation leading to linoleic acid synthesis occurs on phosphatidylcholine located outside the plastids. Monogalactosyl diacylglycerol is therefore synthesized by the coupled pathway, using plastid-derived palmitic acid and endoplasmic reticulum-derived linoleic acid. These results highlight essential differences in lipid biosynthetic pathways between the red algae and the green lineage, which includes plants and green algae. PMID- 17416898 TI - Eap1p, an adhesin that mediates Candida albicans biofilm formation in vitro and in vivo. AB - Candida albicans is the leading cause of systemic fungal infections in immunocompromised humans. The ability to form biofilms on surfaces in the host or on implanted medical devices enhances C. albicans virulence, leading to antimicrobial resistance and providing a reservoir for infection. Biofilm formation is a complex multicellular process consisting of cell adhesion, cell growth, morphogenic switching between yeast form and filamentous states, and quorum sensing. Here we describe the role of the C. albicans EAP1 gene, which encodes a glycosylphosphatidylinositol-anchored, glucan-cross-linked cell wall protein, in adhesion and biofilm formation in vitro and in vivo. Deleting EAP1 reduced cell adhesion to polystyrene and epithelial cells in a gene dosage dependent manner. Furthermore, EAP1 expression was required for C. albicans biofilm formation in an in vitro parallel plate flow chamber model and in an in vivo rat central venous catheter model. EAP1 expression was upregulated in biofilm-associated cells in vitro and in vivo. Our results illustrate an association between Eap1p-mediated adhesion and biofilm formation in vitro and in vivo. PMID- 17416899 TI - Molecular phylogenetics of Candida albicans. AB - We analyzed data on multilocus sequence typing (MLST), ABC typing, mating type like locus (MAT) status, and antifungal susceptibility for a panel of 1,391 Candida albicans isolates. Almost all (96.7%) of the isolates could be assigned by MLST to one of 17 clades. eBURST analysis revealed 53 clonal clusters. Diploid sequence type 69 was the most common MLST strain type and the founder of the largest clonal cluster, and examples were found among isolates from all parts of the world. ABC types and geographical origins showed statistically significant variations among clades by univariate analysis of variance, but anatomical source and antifungal susceptibility data were not significantly associated. A separate analysis limited to European isolates, thereby minimizing geographical effects, showed significant differences in the proportions of isolates from blood, commensal carriage, and superficial infections among the five most populous clades. The proportion of isolates with low antifungal susceptibility was highest for MAT homozygous a/a types and then alpha/alpha types and was lowest for heterozygous a/alpha types. The tree of clades defined by MLST was not congruent with trees generated from the individual gene fragments sequenced, implying a separate evolutionary history for each fragment. Analysis of nucleic acid variation among loci and within loci supported recombination. Computational haplotype analysis showed a high frequency of recombination events, suggesting that isolates had mixed evolutionary histories resembling those of a sexually reproducing species. PMID- 17416900 TI - Stable preanaphase spindle positioning requires Bud6p and an apparent interaction between the spindle pole bodies and the neck. AB - Faithful partitioning of genetic material during cell division requires accurate spatial and temporal positioning of nuclei within dividing cells. In Saccharomyces cerevisiae, nuclear positioning is regulated by an elegant interplay between components of the actin and microtubule cytoskeletons. Regulators of this process include Bud6p (also referred to as the actin interacting protein Aip3p) and Kar9p, which function to promote contacts between cytoplasmic microtubule ends and actin-delimited cortical attachment points. Here, we present the previously undetected association of Bud6p with the cytoplasmic face of yeast spindle pole bodies, the functional equivalent of metazoan centrosomes. Cells lacking Bud6p show exaggerated movements of the nucleus between mother and daughter cells and display reduced amounts of time a given spindle pole body spends in close association with the neck region of budding cells. Furthermore, overexpression of BUD6 greatly enhances interactions between the spindle pole body and mother-bud neck in a spindle alignment defective dynactin mutant. These results suggest that association of either spindle pole body with neck components, rather than simply entry of a spindle pole body into the daughter cell, provides a positive signal for the progression of mitosis. We propose that Bud6p, through its localization at both spindle pole bodies and at the mother-bud neck, supports this positive signal and provides a regulatory mechanism to prevent excessive oscillations of preanaphase nuclei, thus reducing the likelihood of mitotic delays and nuclear missegregation. PMID- 17416901 TI - Trypanosoma brucei encodes a bifunctional capping enzyme essential for cap 4 formation on the spliced leader RNA. AB - The 5' end of kinetoplastid mRNA possesses a hypermethylated cap 4 structure, which is derived from standard m7GpppN (cap 0) with additional methylations at seven sites within the first four nucleosides on the spliced leader RNA. In addition to TbCe1 guanylyltransferase and TbCmt1 (guanine N-7) methyltransferase, Trypanosoma brucei encodes a second cap 0 forming enzyme. TbCgm1 (T. brucei cap guanylyltransferase-methyltransferase) is a novel bifunctional capping enzyme consisting of an amino-terminal guanylyltransferase domain and a carboxyl terminal methyltransferase domain. Recombinant TbCgm1 transfers the GMP to spliced leader RNA (SL RNA) via a covalent enzyme-GMP intermediate, and methylates the guanine N-7 position of the GpppN-terminated RNA to form cap 0 structure. The two domains can function autonomously in vitro. TbCGM1 is essential for parasite growth. Silencing of TbCGM1 by RNA interference increased the abundance of uncapped SL RNA and lead to accumulation of hypomethylated SL RNA. In contrast, silencing of TbCE1 and TbCMT1 did not affect parasite growth or SL RNA capping. We conclude that TbCgm1 specifically cap SL RNA, and cap 0 is a prerequisite for subsequent methylation events leading to the formation of mature SL RNA. PMID- 17416902 TI - The DinG protein from Escherichia coli is a structure-specific helicase. AB - The Escherichia coli DinG protein is a DNA damage-inducible member of the helicase superfamily 2. Using a panel of synthetic substrates, we have systematically investigated structural requirements for DNA unwinding by DinG. We have found that the helicase does not unwind blunt-ended DNAs or substrates with 3'-ss tails. On the other hand, the 5'-ss tails of 11-15 nucleotides are sufficient to initiate DNA duplex unwinding; bifurcated substrates further facilitate helicase activity. DinG is active on 5'-flap structures; however, it is unable to unwind 3'-flaps. Similarly to the homologous Saccharomyces cerevisiae Rad3 helicase, DinG unwinds DNA.RNA duplexes. DinG is active on synthetic D-loops and R-loops. The ability of the enzyme to unwind D-loops formed on superhelical plasmid DNA by the E. coli recombinase RecA suggests that D-loops may be natural substrates for DinG. Although the availability of 5'-ssDNA tails is a strict requirement for duplex unwinding by DinG, the unwinding of D-loops can be initiated on substrates without any ss tails. Since DinG is DNA damage inducible and is active on D-loops and forked structures, which mimic intermediates of homologous recombination and replication, we conclude that this helicase may be involved in recombinational DNA repair and the resumption of replication after DNA damage. PMID- 17416903 TI - Peroxisome proliferator activator receptor gamma coactivator-1 expression is reduced in obesity: potential pathogenic role of saturated fatty acids and p38 mitogen-activated protein kinase activation. AB - Peroxisome proliferator activator receptor-gamma coactivator 1 (PGC-1) is a major candidate gene for diabetes-related metabolic phenotypes, contributing to decreased expression of nuclear-encoded mitochondrial genes in muscle and adipose tissue. We have demonstrated that muscle expression of PGC-1alpha and -beta is reduced in both genetic (Lep(ob)/Lep(ob)) and acquired obesity (high fat diet). In C57BL6 mice, muscle PGC-1alpha expression decreased by 43% (p < 0.02) after 1 week of a high fat diet and persisted more than 11 weeks. In contrast, PGC-1alpha reductions were not sustained in obesity-resistant A/J mice. To identify mediators of obesity-linked reductions in PGC-1, we tested the effects of cellular nutrients in C2C12 myotubes. Although overnight exposure to high insulin, glucose, glucosamine, or amino acids had no effect, saturated fatty acids potently reduced PGC-1alpha and -beta mRNA expression. Palmitate decreased PGC-1alpha and -beta expression by 38% (p = 0.01) and 53% (p = 0.006); stearate similarly decreased expression of PGC-1alpha and -beta by 22% (p = 0.02) and 39% (p = 0.02). These effects were mediated at a transcriptional level, as indicated by an 11-fold reduction of PGC-1alpha promoter activity by palmitate and reversal of effects by histone deacetylase inhibition. Palmitate also (a) reduced expression of tricarboxylic acid cycle and oxidative phosphorylation mitochondrial genes and (b) reduced oxygen consumption. These effects were reversed by overexpression of PGC-1alpha or -beta, indicating PGC-1 dependence. Palmitate effects also required p38 MAPK, as demonstrated by 1) palmitate-induced increase in p38 MAPK phosphorylation, 2) reversal of palmitate effects on PGC-1 and mitochondrial gene expression by p38 MAPK inhibitors, and 3) reversal of palmitate effects by small interfering RNA-mediated decreases in p38alpha MAPK. These data indicate that obesity and saturated fatty acids decrease PGC-1 and mitochondrial gene expression and function via p38 MAPK-dependent transcriptional pathways. PMID- 17416904 TI - DNase X is a glycosylphosphatidylinositol-anchored membrane enzyme that provides a barrier to endocytosis-mediated transfer of a foreign gene. AB - DNase X is the first mammalian DNase to be isolated that is homologous to DNase I. In this study, we have examined its function using a novel monoclonal antibody and showed it to be expressed on the cell surface as a glycosylphosphatidylinositolanchored membrane protein. High level expression was observed in human muscular tissues and in myotubes obtained in vitro from RD rhabdomyosarcoma cells. We observed that RD myotubes incorporated a foreign gene, lacZ, by endocytosis but that expression of the encoded coding product, beta galactosidase, was strongly inhibited. Overexpression of DNase X inhibited endocytosis-mediated gene transfer, whereas knockdown of DNase X with small interfering RNA had the opposite effect. These results reveal that DNase X provides a cell surface barrier to endocytosis-mediated gene transfer. PMID- 17416905 TI - Effects of in vitro oocyte maturation and embryo culture on the expression of glucose transporters, glucose metabolism and insulin signaling genes in rhesus monkey oocytes and preimplantation embryos. AB - Glucose plays a fundamental role during oogenesis and embryogenesis, satisfying the metabolic demands of oocytes and embryos, providing for stored energy reserves in the form of glycogen and supporting nucleotide biosynthesis via the pentose phosphate pathway. Glucose also contributes to the production of amino acids, glycosylated proteins and extracellular components. A detailed understanding of the molecular mechanisms that mediate and regulate glucose uptake and metabolism at different stages of oogenesis and preimplantation embryogenesis could greatly benefit the development of improved methods for in vitro oocyte maturation and in vitro embryo production. Although these processes have been examined in a variety of rodent and agricultural species, detailed information has not yet been described for non-human primates. In this study, we examined the expression of the genes encoding glucose transporters, glucose metabolism enzymes and potential regulators of glucose metabolism in rhesus monkey oocytes and embryos. The data reveal stage-specific regulation of expression of specific types of glucose transporters, stage-specific changes in expression of genes related to different pathways of glucose metabolism and temporal changes in the expression of mRNAs related to insulin signaling. Additionally, the data reveal significant differences in expression of some of these genes in cultured embryos as compared with flushed embryos and between oocytes and embryos obtained following different hormonal stimulation and oocyte maturation protocols. PMID- 17416906 TI - Multicentre evaluation of the adenosine agonist GR79236X in patients with dental pain after third molar extraction. AB - BACKGROUND: Adenosine is analgesic in humans, and the selective adenosine A1 receptor agonist GR79236X has significant anti-nociceptive activity in an animal pain model of inflammatory pain. METHODS: Seventy-nine patients with moderate pain after third molar extraction under general anaesthesia were randomized to receive a 15 min double-blind infusion containing either GR79236X 4 microg kg-1, GR79236X 10 microg kg-1, diclofenac 50 mg, or saline placebo. Rescue analgesia was promptly available to all patients. RESULTS: Meaningful pain relief (mild or no pain) was attained by 9 (47%) patients in the placebo group, 12 (63%) patients in the GR79236 4 microg kg-1 group, 10 (48%) patients in the 10 microg kg-1 group, and 16 (80%) patients in the diclofenac 50 mg group. Neither dose of GR79236 produced a significant improvement over placebo, but diclofenac was superior to both placebo (P=0.036) and GR79236 10 microg kg-1 (P=0.034). Median times to rescue or additional analgesia were 62, 100, 60, and 363 min for patients receiving placebo, GR79236 4 microg kg-1, 10 microg kg-1, and diclofenac 50 mg, respectively (diclofenac significantly longer than placebo, P=0.002 log rank test). Pain control was poor in the placebo group and in both GR79236 groups, with between 79 and 86% of patients having good pain control (i.e. mild or no pain) for <20% of the time compared with only 30% of patients who received diclofenac. CONCLUSION: We found no evidence of efficacy of GR79236 compared with placebo, but the active control diclofenac was effective. It is possible that a higher dose of GR79236 might have been effective or that i.v. administration of this drug does not achieve appropriate concentrations in the brain or peripheral nerves. PMID- 17416907 TI - Oral clonidine vs midazolam in the prevention of sevoflurane-induced agitation in children. a prospective, randomized, controlled trial. AB - BACKGROUND: This randomized, double-blind study tested the hypothesis that, in comparison with midazolam, premedication with oral clonidine reduces the incidence of emergence agitation in preschool children anaesthetized with sevoflurane. METHODS: Sixty-eight ASA I-II children undergoing circumcision were randomized into three groups to receive different oral premedication given 30 min before anaesthesia: midazolam 0.5 mg kg-1, clonidine 2 microg kg-1, and clonidine 4 microg kg-1. Sevoflurane anaesthesia was administered via a facemask (O2/N2O: 40/60). Analgesia was with penile block (bupivacaine 0.5% 0.3 ml kg-1) and rectal paracetamol (30 mg kg-1). During the first postoperative hour, children were evaluated using a modified 'objective pain scale'. RESULTS: Only the 4 microg kg 1 dose of clonidine was associated with a significant reduction in emergence agitation. Fewer children in the clonidine 4 microg kg-1 group displayed agitation (25%) than in the midazolam group (60%) (P=0.025). Incidence of hypotension and bradycardia, time to first micturition and first drink did not differ among groups. CONCLUSIONS: In comparison with midazolam, clonidine 4 microg kg-1 reduced sevoflurane-induced emergence agitation without increasing postoperative side-effects. PMID- 17416908 TI - Removal of the laryngeal tube in children: anaesthetized compared with awake. AB - BACKGROUND: Laryngeal tube (LT) is a useful airway device in children, but there is no objective evidence that removal of LT in awake state is better than in anaesthetized state. So, we compared the incidence of respiratory adverse events after the removal of LT, either under anaesthesia or on awakening. METHODS: Seventy healthy children between 1 and 12 yr of age were enrolled in this study. Anaesthesia was induced and maintained with sevoflurane. After induction of anaesthesia, patients were randomized into two groups: removal of LT in anaesthetized state (Group A: 2% sevoflurane) and in awake state (Group B). During and within 1 min of the removal of LT, airway complications such as upper airway obstruction, cough, vomiting, teeth clenching, hypersalivation, desaturation <90%, and laryngospasm were recorded. RESULTS: Cough (37.1 vs 2.9%), hypersalivation (28.6 vs 5.7%), desaturation (20 vs 0%), and LT dislocation during emergence relating to the patient's movement (26.5 vs 0%) occurred more frequently in Group B (P < 0.05). Upper airway obstruction occurred more frequently (68.6 vs 31.4%) in Group A, and it was easily resolved by chin or jaw lifting. CONCLUSION: LT removal in anaesthetized state reduced cough, hypersalivation, and prevented tube displacement and hypoxia. Upper airway obstruction in the anaesthetized state should be predicted and managed with chin or jaw lifting. PMID- 17416909 TI - Is there an anatomical substrate for idiopathic paroxysmal atrial fibrillation? A case-control echocardiographic study. AB - AIM: Idiopathic paroxysmal atrial fibrillation (AF) occurs in patients with apparently normal heart. Its mechanisms may be complex and are poorly understood. The aim of the study was to evaluate whether patients with idiopathic AF have any structural abnormality that may explain the occurrence of AF. METHODS AND RESULTS: A case-control study was undertaken including 60 consecutive patients (mean age 48 +/- 12 years; 75% men) with idiopathic AF admitted to the emergency department. Sixty sex- and age-matched healthy volunteers made up the control group. An echocardiogram was performed in all patients and volunteers to assess the left atrial (LA) and ventricular (LV) dimensions and valvular function. LV diastolic function was also evaluated by analysis of the LV inflow and pulmonary vein flow velocity patterns and tissue Doppler imaging of the mitral annulus. All AF patients showed normal echocardiographic studies with similar LV dimensions, ejection fraction, and diastolic function when compared with normal controls. However, patients with AF had larger LA dimensions (27 +/- 3 vs. 24 +/- 3 mm/m(2)), LA area (10 +/- 2 vs. 8 +/- 2 mm(2)/m(2)), and LA volume (27 +/- 9 vs. 19 +/- 6 mL/m(2)) (P < 0.05 for all). Among patients with AF, there were no differences in LA size between patients with a first episode or recurrent paroxysmal episodes. CONCLUSION: Patients with idiopathic AF showed larger left atria when compared with controls, there being no differences between patients with a first episode or a recurrence. This suggests the presence of an enhanced substrate to develop idiopathic lone AF. PMID- 17416910 TI - Transvenous cryo-ablation of the slow pathway for the treatment of atrioventricular nodal re-entrant tachycardia: a single-centre initial experience study. AB - AIMS: Within the last several years, transvenous cryo-ablation has become an alternative method to perform ablation of the slow-pathway. This study evaluated the acute and long-term safety and effectiveness of atrio-ventricular nodal re entrant tachycardia (AVNRT) cryo-ablation. METHODS AND RESULTS: The first 69 consecutive patients with AVNRT (60 slow-fast, 4 fast-slow, and 5 slow-slow) who underwent slow-pathway cryo-ablation were included. Mean age was 37 +/- 15, body weight 68 +/- 14 kg, symptom duration 125 +/- 104 months, and number of ineffective antiarrhythmic (AA) drugs 1.8 +/- 1.4. A 7 Fr cryo-catheter (Cryocath(A)) was used, with initially 4-mm-tip and later with 6-mm-tip electrode. Cryo-mapping (n = 7.9 +/- 8.4 per pt) was performed at the temperature of -30 degrees C to test the effect on the target ablation site. Successful cryo mapping was defined as abolition of nodal conduction jump or AV nodal refractory period prolongation. Cryo-ablation (n = 5.1 +/- 4.9 per pt) was then applied by freezing to -75 degrees C for 4 min in duration if no AV-block occurred. Acute procedural success (defined as AVNRT non-inducibility) after the first cryo ablation attempt was achieved in 60/69 patients (87%). During cryo-ablation, inadvertent transient AV-block was encountered in 14 patients (five I AV-block and nine II-III AV-block). A mid-septal target site was the only variable correlated with inadvertent AV-block occurrence during cryo-ablation (P < 0.02). Long-term clinical success after cryo-ablation was globally achieved in 56/66 (85%) with a mean follow-up of 18 +/- 9 months (3 pts dropped-out). After the first procedure, 41/66 (62%) had no relapse, eight had a dramatic reduction in AVNRT duration-frequency and considered themselves cured, and five needed previously ineffective AA (with no relapse in three, drastic reduction in AVNRT duration-frequency in two). The five last patients needed one or more procedures, after which one had no recurrence and one had reduction in duration-frequency. Absence of recurrence after the first procedure was positively correlated with 6 mm-tip cryo-catheter use (<0.001) and negatively with acute procedural success (<0.001). At multivariate analysis, both were independently significant (<0.04 and <0.008, respectively). Long-term clinical success was correlated only with 6 mm-tip cryo-catheter use (<0.001). CONCLUSIONS: Slow pathway cryo-ablation is associated with an acute success but a higher recurrence rate. A 6-mm-tip cryo catheter seems to assure during cryo-ablation a large acute and long-term success. AV-block seems non-guaranteed by a negative cryo-mapping, stressing on need of a careful surveillance. Nevertheless, the theoretical advantage of avoiding the risk of permanent AV-block when compared with radiofrequency needs larger series to be demonstrated. PMID- 17416911 TI - Architecture of the pruned tree: impact of contrasted pruning procedures over 2 years on shoot demography and spatial distribution of leaf area in apple (Malus domestica). AB - BACKGROUND AND AIMS: Demography and spatial distribution of shoots are rarely studied on pruned trees. The present 2-year study deals with the effect of pruning strategies on shoot demography and development, and consequences on the spatial distribution of leaf area in three architecturally contrasted - from type II to IV - apple cultivars: 'Scarletspur Delicious', 'Golden Delicious' and 'Granny Smith'. METHODS: All trees were initially subjected during 5 years to Central Leader training with winter heading on all long shoots. For 2 years, half of the trees were further trained with Centrifugal training, where removal of flowering shoots - called extinction pruning - was carried out along the trunk and at the bottom of branches at flowering time. During these 2 years, shoot type (vegetative, inflorescence) and length, and the three-dimensional spatial distribution of all shoots were assessed with an electromagnetic digitizer. KEY RESULTS: Shoot demography, frequency of transitions toward an inflorescence from either an inflorescence (bourse-over-bourse) or a vegetative shoot (trend toward flowering), and the number of bourse-shoots per bourse were strongly affected by cultivar, with little influence of tree manipulation. In contrast, the proportion of vegetative long shoots developing from previous year latent buds was significantly lower in Centrifugal-trained trees for the three cultivars. Canopy volume showed large variations between cultivars, but only that of 'Granny Smith' was affected by tree manipulation in the 2 years. Spatial distribution of shoots varied significantly according to cultivar and manipulation. In 'Scarletspur Delicious' and, to a lesser extent 'Golden Delicious', the distribution of vegetative and flowering shoots in the outer and the inner parts, respectively, was not affected by tree manipulation. In contrast, in 'Granny Smith', vegetative shoots were stimulated in the periphery of Central Leader trees, whereas flowering shoots were stimulated in the periphery of Centrifugal-trained trees. CONCLUSIONS: In apple, the variability of responses to contrasted pruning strategies partly depends on the genetically determined growth and flowering habit of the cultivar. PMID- 17416912 TI - Portrait of the expansin superfamily in Physcomitrella patens: comparisons with angiosperm expansins. AB - BACKGROUND AND AIMS: Expansins are plant cell wall loosening proteins important in a variety of physiological processes. They comprise a large superfamily of genes consisting of four families (EXPA, EXPB, EXLA and EXLB) whose evolutionary relationships have been well characterized in angiosperms, but not in basal land plants. This work attempts to connect the expansin superfamily in bryophytes with the evolutionary history of this superfamily in angiosperms. METHODS: The expansin superfamily in Physcomitrella patens has been assembled from the Physcomitrella sequencing project data generated by the Joint Genome Institute and compared with angiosperm expansin superfamilies. Phylogenetic, motif, intron and distance analyses have been used for this purpose. KEY RESULTS: A gene superfamily is revealed that contains similar numbers of genes as found in arabidopsis, but lacking EXLA or EXLB genes. This similarity in gene numbers exists even though expansin evolution in Physcomitrella diverged from the angiosperm line approx. 400 million years ago. Phylogenetic analyses suggest that there were a minimum of two EXPA genes and one EXPB gene in the last common ancestor of angiosperms and Physcomitrella. Motif analysis seems to suggest that EXPA protein function is similar in bryophytes and angiosperms, but that EXPB function may be altered. CONCLUSIONS: The EXPA genes of Physcomitrella are likely to have maintained the same biochemical function as angiosperm expansins despite their independent evolutionary history. Changes seen at normally conserved residues in the Physcomitrella EXPB family suggest a possible change in function as one mode of evolution in this family. PMID- 17416913 TI - Linking the evolution of gender variation to floral development. AB - BACKGROUND AND AIMS: In the present review, I have endeavoured to conduct a joint assessment of the thinking underlying the evolutionary genetics of gender polymorphism and the developmental genetics of gender determination. It is my hope, through highlighting the historical development of ideas in two related but somewhat disparate sets of scientific literature, to encourage a synthetic perspective that integrates the two. SCOPE: An overview is provided of various theories on the evolution of sex polymorphism and examples of evidence that has been brought to bear in support of them. Current knowledge on floral development is summarized, with an emphasis on gender variation. Finally, an attempt is made to integrate the two perspectives with the hope that it will encourage future research at the interface. CONCLUSIONS: Evolutionary models of gender evolution have, of necessity, posited genetic effects that are relatively simple in their impacts. Emerging insights from developmental genetics have demonstrated that the underlying reality is a more complex matrix of interacting factors. The study of gender variation in plants is poised for significant advance through the integration of these two perspectives. Bringing genomic tools to bear on population-level processes, we may finally develop a comprehensive perspective on the evolution of floral gender. PMID- 17416914 TI - New insights into the functional morphology of the lever mechanism of Salvia pratensis (Lamiaceae). AB - BACKGROUND AND AIMS: The functional morphology of Salvia pratensis flowers was re investigated, after new insights revealed that pollen dispensing is one of the main functions of the staminal lever. In particular, no detailed information was available regarding the process of pollen transfer and the forces arising between the pollen-bearing thecae and the pollinating bee's body. The assumption was made that these forces play a significant role in pollen dispensing. METHODS: The functional morphology of S. pratensis flowers and the interaction between flowers and bees (Apis mellifera) were studied by reconstructing stress and strains by using qualitative and semi-quantitative theoretical analysis. Flowers were manipulated to study the spatial arrangement of the filament and lever, and of the head and proboscis of the visiting bee inside the tube. Photographs and films of bee visits on flowers were used to analyse the interaction of pollinator and staminal lever. KEY RESULTS: The spoon-shaped lower lever of S. pratensis has a small hole through which a bee introduces its proboscis into the corolla tube. Although mentioned for the first time by Kerner von Marilaun in 1891, presented here is the first drawing and the first photograph showing this interaction in detail. The analysis of the interaction of flower visitor and the lever mechanism revealed that the position of bees on different flowers is spatially very similar. Flower morphology constrains postures of legitimately nectar-probing bees within narrow bounds. A theoretical discussion on structural elements and force progression in the flower allows the principles of lightweight architecture in flower morphology to be recognized. CONCLUSIONS: The staminal lever of S. pratensis is a pollen-dispensing device. It seems to influence the amount of pollen deposited on pollinators by determining the forces arising between the pollinator and the pollen. The relevant forces occur either during the first, dynamic phase or during the second, almost static phase of a flower visit. PMID- 17416915 TI - eSET irrespective of the availability of a good-quality embryo in the first cycle only is not effective in reducing overall twin pregnancy rates. AB - INTRODUCTION: In several clinics, elective single-embryo transfer (eSET) is applied in a selected group of patients based on age and the availability of a good-quality embryo. Whether or not eSET can be applied irrespective of the presence of a good-quality embryo in the first cycle, to further reduce the twin pregnancy rate, remains to be elucidated. METHODS: In patients <38 years two transfer strategies were compared, which differed in the first cycle only: group A (n = 141) received eSET irrespective of the availability of a good-quality embryo, and group B (n = 174) received eSET when a good-quality embryo was available while otherwise they received double embryo transfer (DET; referred to as eSET/DET transfer policy). In any subsequent cycle, in both groups the eSET/DET transfer policy was applied. RESULTS: After completion of their IVF treatment (including a maximum of three fresh cycles and the transfer of frozen thawed embryos), comparable cumulative live birth rates (62.4% in group A and 62.6% in group B) and twin pregnancy rates (10.1 versus 13.4%) were found. However, patients in group A required significantly more fresh (2.0 versus 1.8) and frozen (0.8 versus 0.5) cycles. CONCLUSIONS: The transfer of one embryo in the first cycle, irrespective of the availability of a good-quality embryo, in all patients <38 years, is not an effective transfer policy for reducing the overall twin pregnancy rate. PMID- 17416916 TI - Correlation of maternal plasma total cell-free DNA and fetal DNA levels with short term outcome of first-trimester vaginal bleeding. AB - BACKGROUND: The current methods using sonographic parameters and/or maternal serum beta-HCG levels to predict spontaneous abortion are not satisfactory. The aim of this study was to determine whether maternal plasma fetal DNA and total DNA levels could be used to predict spontaneous abortion. METHODS: We prospectively studied pregnant women who presented with vaginal bleeding in the first trimester of pregnancy, and those who had no vaginal bleeding (controls). DYS14 and the beta-globin gene were used to measure the maternal plasma levels of fetal and total DNA, respectively, by real-time PCR. RESULTS: A total of 1114 women were studied. Both maternal plasma fetal and total DNA concentrations increased with gestation from 6 to 11.6 weeks in the controls. The multiple of medians (MoMs) of fetal and total DNA concentration in those who miscarried were significantly greater (P < 0.001) than in the normal controls by about 5- and 4 fold respectively. Using a cut-off value of 1.6 MoMs for total DNA to predict spontaneous abortion, the sensitivity was 98.2% and false positive rate was 4.7%. However, using a cut-off value of 1.8 MoMs for fetal DNA, the corresponding figures were 97% and 44.3%, respectively. CONCLUSIONS: Both maternal plasma fetal and total DNA concentrations increased throughout the first trimester. Significantly high levels of fetal and total DNA were found in those who miscarried. PMID- 17416917 TI - Decision-making about seeking medical advice in an internet sample of women trying to get pregnant. AB - INTRODUCTION: In light of the near universal desire to become a parent one would expect most people to seek medical advice if they were experiencing difficulties conceiving. Yet less than 55% do so and we sought to understand more about this paradox by comparing the psychosocial profile and decision-making of women not yet engaged in the medical process to that of those who had consulted. METHODS: A Fertility Decision-Making Questionnaire was designed and posted on a website dedicated to women trying to conceive. RESULTS: A total of 426 women completed the questionnaire, 56% had not yet consulted a doctor about conceiving (non consulter, NC). Women who had sought treatment had more positive treatment beliefs, and a greater willingness to know if a problem existed, than those who had not yet consulted. Almost 20% of NC women already met the medical definition of infertility and this subgroup had a greater fear of discovering a problem and of being labelled infertile than other women in the study. CONCLUSIONS: Seeking medical advice for fertility problems is mainly associated with what women know or want to know about their fertility and their emotional reactions to that knowledge. Negative reactions can substantially delay seeking help in 20% of women. PMID- 17416918 TI - Decisions for the fate of frozen embryos: fresh insights into patients' thinking and their rationales for donating or discarding embryos. AB - BACKGROUND: In the final decision for the disposition of unused IVF embryos patients must choose between options involving either donation or destruction, and this decision must be made in a context where there is tension about the status of embryos (i.e. whether viewed as potential children or as a base for further development) and whether embryo donation is adoption or tissue donation. This study explored the emotive experience of making a decision for either the destruction or donation of unused embryos. METHODS: Thirty-three patients (9 women and 12 couples) who discarded embryos and 15 (7 women and 4 couples) who donated embryos were interviewed. Interview data were analysed with particular attention to elements of moral deliberation and use of analogy. RESULTS: Adoption and tissue donation metaphors were both identified, and further, a metaphor of pregnancy termination was identified and found to be highly influential in the decision to donate embryos. Contrary to the majority of current evidence, this study found that participants who discarded embryos emphasized the adoption metaphor while embryo donors emphasized the metaphor of pregnancy termination. For each group the decision was driven by awareness of the option they did not want. CONCLUSIONS: The pregnancy termination metaphor emerged as morally relevant and this holds implications for defining and discussing embryo discard in counselling and consent processes. PMID- 17416919 TI - Evaluation of the dynamic responses of female pelvic floor using a novel vaginal probe. AB - The female pelvic floor (PF) provides anatomical support to many visceral organs, such as uterus, bladder, urethra, vagina, and rectum. Physiologically, the PF is made up of a number of highly coordinated muscle groups organized to respond to postural and abdominal stresses to maintain continence. In this article, we describe a new methodology for the evaluation of PF strength using a novel vaginal probe design, having force and displacement sensors. This design was derived on the basis of imaging data showing that force/displacement characteristics are important determinants of the integrity of the PF function. The prototype probe used was constructed to evaluate the dynamic responses to slow voluntary contractions as well as reflex stress contractions. Initial clinical experiments were performed on nine healthy female subjects. The probe recorded the force and displacement signals on the anterior and posterior sides of the subjects' middle vaginal wall in voluntary PF muscle contraction and cough. The time domain and frequency domain characteristics of the dynamic responses, including the force and displacement responses, of the vaginal wall were measured and the power and energy associated with the dynamic responses of the PF were analyzed showing the differences between the dynamic characteristics of the voluntary PF muscle contraction and cough. Results show that voluntary PF muscle contractions have higher amplitudes, longer duration, and higher power than reflex contractions. The design of this probe enables the measurement of force and displacement during rapidly occurring events. PMID- 17416920 TI - Striatal contributions to reward and decision making: making sense of regional variations in a reiterated processing matrix. AB - The striatum is the major input nucleus of the basal ganglia. It is thought to play a key role in learning on the basis of positive reinforcement and in action selection. One view of the striatum conceives it as comprising a reiterated matrix of processing units that perform common operations in different striatal regions, namely synaptic plasticity according to a three-factor rule, and lateral inhibition. These operations are required for reinforcement learning and selection of previously reinforced actions. Analysis of the behavioral effects of circumscribed lesions of the striatum, however, suggests regional specialization of learning and decision-making operations. We consider how a basic processing unit may be modified by regional variations in neurochemical parameters, for example, by the gradient in density of dopamine terminals from dorsal to ventral striatum. These variations suggest subtle differences between dorsolateral and ventromedial striatal regions in the temporal properties of dopamine signaling, which are superimposed on regional differences in connectivity. We propose that these variations make sense in relation to the temporal structure of activity in striatal inputs from different regions, and the requirements of different learning operations. Dorsolateral striatal (DLS) regions may be subject to brief, precisely timed pulses of dopamine, whereas ventromedial striatal regions integrate dopamine signals over a longer time course. These differences may be important for understanding regional variations in the contribution to reinforcement of habits, versus incentive processes that are sensitive to the value of expected rewards. PMID- 17416921 TI - Model-based fMRI and its application to reward learning and decision making. AB - In model-based functional magnetic resonance imaging (fMRI), signals derived from a computational model for a specific cognitive process are correlated against fMRI data from subjects performing a relevant task to determine brain regions showing a response profile consistent with that model. A key advantage of this technique over more conventional neuroimaging approaches is that model-based fMRI can provide insights into how a particular cognitive process is implemented in a specific brain area as opposed to merely identifying where a particular process is located. This review will briefly summarize the approach of model-based fMRI, with reference to the field of reward learning and decision making, where computational models have been used to probe the neural mechanisms underlying learning of reward associations, modifying action choice to obtain reward, as well as in encoding expected value signals that reflect the abstract structure of a decision problem. Finally, some of the limitations of this approach will be discussed. PMID- 17416922 TI - Splitting the difference: how does the brain code reward episodes? AB - Animal research and human brain imaging findings suggest that reward processing involves distinct anticipation and outcome phases. Error terms in popular models of reward learning (such as the temporal difference [TD] model) do not distinguish between the updating of expectations in response to reward cues and outcomes. Thus, correlating a single error term with neural activation assumes recruitment of similar neural substrates at each update. Here, we split the error term to separately model reward prediction and prediction errors, and compare the fit of single versus split error terms to functional magnetic resonance imaging (FMRI) data acquired during a monetary incentive delay task. We speculate and find that while the nucleus accumbens computes gain prediction in response to cues, the mesial prefrontal cortex (MPFC) computes gain prediction errors in response to outcomes. In addition to offering a more comprehensive and anatomically situated view of reward processing, split error terms generate novel predictions about psychiatric symptoms and lesion-induced deficits. PMID- 17416923 TI - Integration of cognitive and motivational information in the primate lateral prefrontal cortex. AB - The prefrontal cortex (PFC), particularly the lateral prefrontal cortex (LPFC), has an important role in cognitive information processing. The area receives projections from sensory association cortices and sends outputs to motor-related areas. Neurons in LPFC code the behavioral significance of stimuli, which can be abstract precursors for complex motor commands and are structured hierarchically. Loss of these neurons leads to a lack of flexibility in decision making, such as seen in stereotyped behaviors. However, to make more appropriate decisions the code for behavioral significance has to reflect the subject's own desires and demands. Indeed, LPFC has connections with reward-related areas, such as the orbitofrontal cortex (OFC), basal ganglia, and medial prefrontal cortex. Recently, many studies have reported reward modulation of neural codes of behavioral significance. Using an asymmetric reward paradigm, we can investigate the functional specificity of LPFC neurons that code both cognitive information and motivational information. In this review, we will discuss details of neuronal properties of LPFC neurons from the viewpoints of cognitive information processing and motivational information processing, and the question of how these two pieces of information are integrated. Abstract coding and contextual representations in the cognitive information processing are functional characteristics of LPFC. Such functional specificity in LPFC cognitive processes is supported by a long-term scale of reward history in the motivational information processing. The integration enables us to make an elaborate decision with respect to goal-directed behavior in complex circumstances. PMID- 17416924 TI - Functional anatomy of the female pelvic floor. AB - The anatomic structures in the female that prevent incontinence and genital organ prolapse on increases in abdominal pressure during daily activities include sphincteric and supportive systems. In the urethra, the action of the vesical neck and urethral sphincteric mechanisms maintains urethral closure pressure above bladder pressure. Decreases in the number of striated muscle fibers of the sphincter occur with age and parity. A supportive hammock under the urethra and vesical neck provides a firm backstop against which the urethra is compressed during increases in abdominal pressure to maintain urethral closure pressures above the rapidly increasing bladder pressure. This supporting layer consists of the anterior vaginal wall and the connective tissue that attaches it to the pelvic bones through the pubovaginal portion of the levator ani muscle, and the uterosacral and cardinal ligaments comprising the tendinous arch of the pelvic fascia. At rest the levator ani maintains closure of the urogenital hiatus. They are additionally recruited to maintain hiatal closure in the face of inertial loads related to visceral accelerations as well as abdominal pressurization in daily activities involving recruitment of the abdominal wall musculature and diaphragm. Vaginal birth is associated with an increased risk of levator ani defects, as well as genital organ prolapse and urinary incontinence. Computer models indicate that vaginal birth places the levator ani under tissue stretch ratios of up to 3.3 and the pudendal nerve under strains of up to 33%, respectively. Research is needed to better identify the pathomechanics of these conditions. PMID- 17416925 TI - Physiology of upward transport in the human female genital tract. AB - The uterus and fallopian tubes represent a functionally united peristaltic pump under the endocrine control of ipsilateral ovary. We have examined this function by using hysterosalpingoscintigraphy (HSS), recording of intrauterine pressure, electrohysterography, and Doppler sonography of the fallopian tubes. An uptake of labeled particles into the uterus was observed during the follicular and luteal phases of the cycle after application into the vagina. Transport into the oviducts, however, could only be demonstrated during the follicular phase. Furthermore, the predominant transport was into the tube ipsilateral to the ovary containing the dominant follicle. The pregnancy rate following spontaneous intercourse or insemination was higher in those women in whom ipsilateral transport could be demonstrated. The amount of material transported to the ipsilateral tube was increased after oxytocin administration, as demonstrated by radionuclide imaging and by Doppler sonography following instillation of ultrasound contrast medium. An increase in the basal tone and amplitude of contractions was observed after oxytocin administration. These results support the idea that the uterus and fallopian tubes act as a peristaltic pump, which increases transport of sperm into the oviduct ipsilateral to the ovary bearing the dominant follicle. Oxytocin appears to play a critical role in this peristaltic pump. A failure of the peristaltic mechanism is possibly responsible for infertility. We propose the term tubal transport disorder (TTD) as a nosological entity. Results from HSS could be a useful adjunct for choosing treatment modalities in patients with patent fallopian tubes suffering from infertility. These patients may be better served with in vitro fertilization (IVF). PMID- 17416926 TI - Biosafety and selectable markers. AB - Francisella tularensis (F. tularensis) is one of the most infectious pathogens known. Although the disease caused by this bacterium is rarely fatal with appropriate antibiotic therapy, it is often severely debilitating. Laboratory work with F. tularensis poses a significant hazard, and it is essential that appropriate laboratory facilities, trained personnel, and suitable working practices are in place in any organization working with this pathogen. Work with human virulent strains should be carried out at containment level 3. Vaccines play an important potential role in the protection of individuals working with F. tularensis, but the live vaccine strain (LVS) vaccine is not currently licensed for use in humans. Because antibiotic therapy is central to the treatment of human tularemia, antibiotic resistance markers for use in genetic manipulation studies should be carefully selected. PMID- 17416927 TI - Fetal blood flow in branching models of the chorionic arterial vasculature. AB - Fetal development depends on adequate exchange of materials between the fetus and maternal circulatory systems, which requires optimal distribution of blood vessels over the chorionic plate to ensure perfusion of the whole placental volume. Based on a previous investigation of the architecture of the chorionic vessels in the human placenta, we developed in this study typical models for the dichotomous and monopodial segments of the chorionic arteries of a mature placenta. Each model also included some intraplacental (IP) vessels that branch off into the cotyledons perpendicular to the chorionic arteries. Computational analysis of steady blood flow through these models was performed to explore the distribution of fetal blood over the chorionic plate. The results demonstrated that energy losses are small in the monopodial model, which explains their efficient delivery of fetal blood over the chorionic plate in cases of a marginal cord insertion. On the other hand, the dichotomous model is efficient in distributing a relatively large volume of blood over large areas near the bifurcation. Accordingly, the combination of dichotomous and monopodial bifurcation in a normal chorionic plate ensures a uniform blood perfusion of the placenta. Simulations with narrow daughter and IP vessels did not result in significant changes in the main mother tubes, supporting clinical observations in which umbilical blood flow remains normal although some peripheral vessels may be occluded. PMID- 17416928 TI - Cost, benefit, tonic, phasic: what do response rates tell us about dopamine and motivation? AB - The role of dopamine in decision making has received much attention from both the experimental and computational communities. However, because reinforcement learning models concentrate on discrete action selection and on phasic dopamine signals, they are silent as to how animals decide upon the rate of their actions, and they fail to account for the prominent effects of dopamine on response rates. We suggest an extension to reinforcement learning models in which response rates are optimally determined by balancing the tradeoff between the cost of fast responding and the benefit of rapid reward acquisition. The resulting behavior conforms well with numerous characteristics of free-operant responding. More importantly, this framework highlights a role for a tonic signal corresponding to the net rate of rewards, in determining the optimal rate of responding. We hypothesize that this critical quantity is conveyed by tonic levels of dopamine, explaining why dopaminergic manipulations exert a global affect on response rates. We further suggest that the effects of motivation on instrumental rates of responding are mediated through its influence on the net reward rate, implying a tight coupling between motivational states and tonic dopamine. The relationships between phasic and tonic dopamine signaling, and between directing and energizing effects of motivation, as well as the implications for motivational control of habitual and goal-directed instrumental action selection, are discussed. PMID- 17416929 TI - New BODIPY lipid probes for fluorescence studies of membranes. AB - Many fluorescent lipid probes tend to loop back to the membrane interface when attached to a lipid acyl chain rather than embedding deeply into the bilayer. To achieve maximum embedding of BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) fluorophore into the bilayer apolar region, a series of sn-2 acyl-labeled phosphatidylcholines was synthesized bearing 4,4-difluoro-1,3,5,7-tetramethyl-4 bora-3a,4a-diaza-s-indacene-8-yl (Me(4)-BODIPY-8) at the end of C(3)-, C(5)-, C(7)-, or C(9)-acyl. A strategy was used of symmetrically dispersing the methyl groups at BODIPY ring positions 1, 3, 5, and 7 to decrease fluorophore polarity. Iodide quenching of the phosphatidylcholine probes in bilayer vesicles confirmed that the Me(4)-BODIPY-8 fluorophore was embedded in the bilayer. Parallax analysis of Me(4)-BODIPY-8 fluorescence quenching by phosphatidylcholines containing iodide at different positions along the sn-2 acyl chain indicated that the penetration depth of Me(4)-BODIPY-8 into the bilayer was determined by the length of the linking acyl chain. Evaluation using monolayers showed minimal perturbation of <10 mol% probe in fluid-phase and cholesterol-enriched phosphatidylcholine. Spectral characterization in monolayers and bilayers confirmed the retention of many features of other BODIPY derivatives (i.e., absorption and emission wavelength maxima near 498 nm and approximately 506-515 nm) but also showed the absence of the 620-630 nm peak associated with BODIPY dimer fluorescence and the presence of a 570 nm emission shoulder at high Me(4) BODIPY-8 surface concentrations. We conclude that the new probes should have versatile utility in membrane studies, especially when precise location of the reporter group is needed. PMID- 17416930 TI - p53R2-dependent ribonucleotide reduction provides deoxyribonucleotides in quiescent human fibroblasts in the absence of induced DNA damage. AB - Human fibroblasts in culture obtain deoxynucleotides by de novo ribonucleotide reduction or by salvage of deoxynucleosides. In cycling cells the de novo pathway dominates, but in quiescent cells the salvage pathway becomes important. Two forms of active mammalian ribonucleotide reductases are known. Each form contains the catalytic R1 protein, but the two differ with respect to the second protein (R2 or p53R2). R2 is cell cycle-regulated, degraded during mitosis, and absent from quiescent cells. The recently discovered p53-inducible p53R2 was proposed to be linked to DNA repair processes. The protein is not cell cycle-regulated and can provide deoxynucleotides to quiescent mouse fibroblasts. Here we investigate the in situ activities of the R1-p53R2 complex and two other enzymes of the de novo pathway, dCMP deaminase and thymidylate synthase, in confluent quiescent serum-starved human fibroblasts in experiments with [5-(3)H]cytidine, [6 (3)H]deoxycytidine, and [C(3)H(3)]thymidine. These cells had increased their content of p53R2 2-fold and lacked R2. From isotope incorporation, we conclude that they have a complete de novo pathway for deoxynucleotide synthesis, including thymidylate synthesis. During quiescence, incorporation of deoxynucleotides into DNA was very low. Deoxynucleotides were instead degraded to deoxynucleosides and exported into the medium as deoxycytidine, deoxyuridine, and thymidine. The rate of export was surprisingly high, 25% of that in cycling cells. Total ribonucleotide reduction in quiescent cells amounted to only 2-3% of cycling cells. We suggest that in quiescent cells an important function of p53R2 is to provide deoxynucleotides for mitochondrial DNA replication. PMID- 17416931 TI - Cytochrome b pseudogene originated from a highly divergent mitochondrial lineage in genus Rupicapra. AB - We have identified a nuclear pseudogene (numt) of cytochrome b (cytb) in chamois. The comparison of a fragment of 402 nucleotides of cytb and the pseudogene between the 2 species Rupicapra rupicapra and Rupicapra pyrenaica allowed direct measurement of relative rates and patterns of evolution. Mitochondrial genes evolved 7 to 12 times faster than their nuclear counterparts. Substitutions in the nucleus include a frameshift and a stop codon. Phylogenetic analysis of nuclear and mitochondrial lineages on Rupicapra and related species showed that the nuclear branch evolved as a functional mitochondrial gene until the split of the 2 species of chamois and as a typical pseudogene later on. We propose that the pseudogene originated from a highly divergent mitochondrial lineage that did not persist in the mitochondrion and transposed to the nucleus in a time close to speciation. The concurrence of highly differentiated lineages at speciation points to hybridization between highly divergent populations. PMID- 17416932 TI - Meiotic restitution in wheat polyhaploids (amphihaploids): a potent evolutionary force. AB - Polyploidy is well recognized as a major force in plant speciation. Among the polyploids in nature, allopolyploids are preponderant and include important crop plants like bread wheat, Triticum aestivum L. (2n = 6x = 42; AABBDD genomes). Allopolyploidy must result through concomitant or sequential events that entail interspecific or intergeneric hybridization and chromosome doubling in the resultant hybrids. To gain insight into the mechanism of evolution of wheat, we extracted polyhaploids of 2 cultivars, Chinese Spring (CS) and Fukuhokomugi (Fuko), of bread wheat by crossing them with maize, Zea mays L. ssp. mays. The derived Ph1-polyhaploids (2n = 3x = 21; ABD) showed during meiosis mostly univalents, which produced first-division restitution (FDR) nuclei that in turn gave rise to unreduced (2n) male gametes with 21 chromosomes. The haploids on maturity set some viable seed. The mean number of seeds per spike was 1.45 +/- 0.161 in CS and 2.3 +/- 0.170 in Fuko. Mitotic chromosome preparations from root tips of the derived plantlets revealed 2n = 42 chromosomes, that is, twice that of the parental polyhaploid, which indicated that they arose by fusion of unreduced male and female gametes formed by the polyhaploid. The Ph1-induced univalency must have produced 2n gametes and hence bilateral sexual polyploidization and reconstitution of disomic bread wheat. These findings highlight the quantum jump by which bread wheat evolved from durum wheat in nature. Thus, bread wheat offers an excellent example of rapid evolution by allopolyploidy. In the induced polyhaploids (ABD) that are equivalent of amphihaploids, meiotic phenomena such as FDR led to regeneration of parental bread wheat, perhaps a simulation of the evolutionary steps that occurred in nature at the time of the origin of hexaploid wheat. PMID- 17416933 TI - Mitochondrial DNA diversity and population structure among southern right whales (Eubalaena australis). AB - The population structure and mitochondrial (mt) DNA diversity of southern right whales (Eubalaena australis) are described from 146 individuals sampled on 4 winter calving grounds (Argentina, South Africa, Western Australia, and the New Zealand sub-Antarctic) and 2 summer feeding grounds (South Georgia and south of Western Australia). Based on a consensus region of 275 base pairs of the mtDNA control region, 37 variable sites defined 37 unique haplotypes, of which only one was shared between regional samples of the Indo-Pacific and South Atlantic Oceans. Phylogenetic reconstruction of the southern right whale haplotypes revealed 2 distinct clades that differed significantly in frequencies between oceans. An analysis of molecular variance confirmed significant overall differentiation among the 4 calving grounds at both the haplotype and the nucleotype levels (F(ST) = 0.159; Phi(ST) = 0.238; P < 0.001). Haplotype diversity was significantly lower in the Indo-Pacific (h = 0.701 +/- 0.037) compared with the South Atlantic (h = 0.948 +/- 0.013), despite a longer history of exploitation and larger catches in the South Atlantic. In fact, the haplotype diversity in the Indo-Pacific basin was similar to that of the North Atlantic right whale that currently numbers about 300 animals. Multidimensional scaling of genetic differentiation suggests that gene flow occurred primarily between adjacent calving grounds within an ocean basin, with mixing of lineages from different calving grounds occurring on feeding grounds. PMID- 17416934 TI - Building character. PMID- 17416935 TI - Pediatric cataract. PMID- 17416936 TI - Vitreoretinal surgery in the ambulatory surgery center in the United States. AB - There is a growing trend in the ambulatory surgery centers of the United States to perform vitreoretinal cases. Logistic and financial considerations affect health care delivery systems worldwide. This article presents the barriers that prevent or impede the performance of posterior segment surgery in the ambulatory surgery center environment and also lists some of the advantages. PMID- 17416937 TI - Role of malondialdehyde and superoxide dismutase in cataractogenesis. AB - Malondialdehyde (MDA) and superoxide dismutase (SOD) in lens and plasma of patients with age-related and myopic cataract were assessed. Higher levels of lens MDA in myopic patients showed a greater lipid peroxidation process than in patients with age-related cataract indicating higher oxidative stress in myopic lens. The difference in plasma MDA of myopic cataract and age-related cataract patients was not significant. The SOD level was lower in myopic patients than in patients with age-related cataract. PMID- 17416938 TI - Visual acuity in central and branch vein retinal occlusion in the presence of macular edema: 1 year of follow-up. AB - One year after laser treatment of 117 eyes with central or branch retinal vein occlusion in the presence of macular edema, visual acuity was equally improved in the central retinal vein occlusion laser-treated and branch retinal vein occlusion non-laser-treated eyes. We concluded that fluorangiographically evident ischemic areas or significant macular edema are indications for laser therapy, and that a poor long-term functional prognosis is always related to more severe macular damage at the onset. PMID- 17416939 TI - A comparison of latanoprost monotherapy with a combination therapy of timolol/dorzolamide in patients with primary open-angle glaucoma. AB - We compared latanoprost monotherapy therapy with timolol/ dorzolamide in patients with primary open-angle glaucoma to evaluate the effects on intraocular pressure (IOP) and occurrence of adverse events. IOP and topical side effects were evaluated at the beginning, first, and third months. Mean IOP was decreased at the third month. The most common side effect was hyperemia (43.6%). We concluded that latanoprost reduces IOP better than fixed combination and its topical side effects are tolerable. PMID- 17416940 TI - Corneal and conjunctival impression cytology in soft contact lens wearers. AB - This study assessed changes in corneal and conjunctional cytology in 25 soft contact lens (CL) wearers and compared them with matched controls. We found 80% of grade IV keratinization in CL users for more than 7 years. Keratinization was directly proportional to duration of CL use. Conjunctival cytological changes worsened with decreasing pH, whereas corneal keratinization showed no significant correlation. PMID- 17416941 TI - Optical coherence tomography following photodynamic therapy with verteporfin for subfoveal predominantly classic choroidal neovascularization. AB - We characterized the changes in optical coherence tomography following photodynamic therapy (PDT) for subfoveal predominantly classic choroidal neovascularization (CNV) in 26 eyes before, 1 week after, and 3 months after treatment. There appears to be a temporal decrease in central retinal thickness, increase in central retinal pigment epithelium/CNV complex thickness, and decrease in subretinal fluid following PDT, making it a useful adjunct in the management of CNV due to of age-related macular degeneration. PMID- 17416942 TI - Early vs late intraocular pressure following trabeculectomy with releasable suture in advanced glaucoma. AB - We retrospectively studied long-term outcome after trabeculectomy with releasable suture in advanced glaucoma in 67 consecutive glaucomic eyes. Intraocular pressure (IOP) was recorded preoperatively and periodically following surgery for up to 2 years. Early (<1 month) postoperative IOP was a reasonable indicator of long-term IOP control in eyes with advanced glaucoma. PMID- 17416943 TI - Orbital cellulitis and irreversible visual loss owing to acute sinusitis. AB - We report a case with orbital cellulitis and irreversible visual loss caused by asymptomatic acute sinusitis in a patient without symptoms or history of sinusitis. The prompt use of intravenous antibiotics in combination with early surgical drainage may prevent irreversible visual loss in the affected eye, and threatened visual loss in the fellow eye can be reduced. PMID- 17416944 TI - Bilateral combined cilioretinal artery and central retinal vein occlusion. AB - A rare case with bilateral combined cilioretinal artery and central retinal vein occlusion (CRVO) is reported in a 45-year-old man. Work-up was positive only for already-present type 2 diabetes. The patient first developed ischemic-type CRVO and cilioretinal artery occlusion in his right eye, and underwent successful trabeculectomy with mitomycin-C following panretinal and grid macular photocoagulation. The fellow eye developed a nonischemic type of CRVO and cilioretinal artery occlusion 4 years after the first episode with good visual outcome. PMID- 17416945 TI - Hemi-retinal vein occlusion following LASIK. AB - We report a case of hemi-retinal vein occlusion following laser-assisted in situ keratomileusis (LASIK) in a healthy 46-year-old Caucasian male. A hemi-retinal vein occlusion following LASIK could be coincidental. However, young age, absence of risk factors and negative laboratory testing require consideration of a causal relationship. PMID- 17416946 TI - Carcinoma ex pleomorphic adenoma of the lacrimal gland: a case report. AB - Carcinoma ex pleomorphic adenoma affects glandular structures, occurring mostly in major salivary glands and less commonly in the lacrimal gland. We present a rare case in the lacrimal gland. We highlight the importance of keeping this rare tumor in mind and the early detection of symptoms because such a malignant transformation could occur in pleomorphic adenomas, with the proportion dependent on the duration of the tumor. PMID- 17416947 TI - Unusual intraorbital foreign body: a case report. AB - Penetrating eye injury is an important cause of ocular morbidity in India, especially in the pediatric and adolescent age group. Sometimes these injuries are associated with retained intraorbital foreign body. We report herein for the first time a child who had an open globe injury with a retained unusually large intraorbital foreign body (iron nut). PMID- 17416948 TI - Bilateral full-thickness macular hole: a case report. AB - Bilateral full-thickness macular hole is a rare retinal disease. A 63-year-old man complained of metamorphopsy on his left eye. Pars plana vitrectomy was performed for stage 2 macular hole with anatomical and functional success. Postoperatively, the right eye showed the same symptoms, and stage 2 macular hole, detected by ocular coherency tomography, developed within 4 months, but closed after pars plana vitrectomy. PMID- 17416949 TI - A case of cystoid macular edema treated by intravitreal triamcinolone in a patient with systemic lupus erythematosis. AB - We describe the use of intravitreal triamcinolone in the treatment of macular edema associated with systematic lupus erythematosus providing short-term resolution of cystoid macular edema and increase in visual acuity. No side effects from the injection have been seen as yet. We suggest this treatment to be considered in patients for whom laser treatment may be inappropriate or not feasible, or even as an alternative/safer option for all patients. PMID- 17416950 TI - Scientific revolution. AB - Has science truly reached its ultimate peak? Some may say that it has, and that our current knowledge base is as good as it is going to get. If we look closely, we may see that we are not standing on the peak, but rather on the edge of a grand precipice that overlooks a future filled with infinite possibilities. There is so much in the universe, and even on our own planet, that we still do not understand. How we choose to validate new knowledge will largely determine how future technologies will be developed for the benefit of man. Perhaps all fields of medicine are still in an infantile stage. There is so much that we have learned, but think of how much more there is to understand. For example, the project called the Fugo Bladetrade mark is now penetrating into multiple fields of medicine outside of ophthalmology. It employs plasma fields to perform surgery. Plasma technology may dramatically revolutionize the way surgery is performed. Rather than cutting or burning tissue, plasma allows us to ablate tissue using very low powers and even allows for portable systems. Plasma offers clean incisions with non-cauterizing hemostasis and less swelling. Other changes in the field of ophthalmology are dramatic. Dr. Daljit Singh has introduced decompression of aqueous from the posterior chamber of the eye, which is an immense step forward for glaucoma. These advances ultimately benefit the patient. Yet, because no major corporate machine owns the rights to this technology, it is often difficult to get the message out. PMID- 17416951 TI - Corneal disk detachment. PMID- 17416952 TI - Orbicularis plication for ptosis: a third alternative. AB - The surgery involves exposure of the orbicularis oculi muscle via a skin flap starting near the upper orbital margin then progressing downward. The orbicularis oculi fibers near the lid margin are then joined to the proximal orbicularis fibers and the skin flap is sutured back to normal position. Over 9 years, 265 operations have been performed on a wide variety and severity of ptosis. The technique is a viable addition/alternative to the existing methods of ptosis surgery. PMID- 17416953 TI - Estimating intraocular pressure using a glass rod. AB - Four masked examiners studied the usefulness of the glass-rod for estimating intraocular pressure (IOP) in a living, anesthetized rabbit eye manometrically set to 135 different random values. With training, the average error in IOP estimation improved from 7.42 +/- 5.56 mmHg to 5.13 +/- 4.48 mmHg. Gentle palpation of the corneal apex with a glass-rod provides tactile and visual clues that may assist in estimating the IOP. PMID- 17416954 TI - Subtenon lidocaine vs topical proparacaine in adult strabismus surgery. AB - Intraoperative subtenon 2% lidocaine and topical 0.5% proparacaine in patients undergoing strabismus surgery were compared. No additional systemic analgesics and sedatives were used. Mean and total pain scores intraoperatively and postoperatively were not different. Each anesthetic agent provides good intraoperative anesthesia and postoperative analgesia. Topical 0.5% proparacaine may be preferred because of its easy administration and fewer side effects, lack of hospital admission, and immediate and predictable alignment of the eyes. PMID- 17416955 TI - Surgical approaches used in the reconstruction of the eyelids after excision of malignant tumors. AB - The choice of reconstructive technique for eyelid defects after malignant tumors was based on tumor location and defect size after tumor excision. Treatments included direct closure for defects measuring less than 25%, Tenzel flap for defects measuring 25-50%, lid-sharing procedures for defects measuring more than 50%, skin grafts for tumors affecting the inner canthus advancement flap for outer canthus tumors, and orbital exenteration for malignant tumors demonstrating orbital invasion. PMID- 17416956 TI - Goniotomy ab interno "a glaucoma filtering surgery" using the Fugo Plasma Blade. AB - Sixteen eyes with absolute glaucoma were treated and followed up for a minimum of six months. Goniotomy ab interno using the Fugo Blade was found to be a safe alternative to conventional trabeculectomy, which safely and effectively reduced intraocular pressure in more than 80% of cases. PMID- 17416957 TI - Conjunctival flaps for pterygium surgery. AB - The literature reveals that pterygium surgery focuses on adjunctive therapy such as mitomycin C and 5-fluorouracil, and beta irradiation with bare sclera technique to decrease recurrence. The following description of the basic surgical technique of conjunctival flaps shows that it is simple, easier to perform, has faster healing time, and is more effective in preventing recurrence than any of the others. PMID- 17416958 TI - Delayed foveal reattachment in scleral buckle surgery for inferior retinal detachment. AB - The study evaluates the prevalence and course of subfoveal serous detachment (SSD) following successful retinal reattachment surgery. SSD was commonly seen in postprimary scleral buckle eyes for inferior rhegmatogenous retinal detachment vs none in the primary pars plana vitrectomy group, irrespective of the break localization. PMID- 17416959 TI - Oxidants and ocular tumors. AB - Serum superoxide dismutase and catalase assays were performed using spectrophotometry in 60 adults and children with benign or malignant tumors and in controls. There was a statistically significant difference in the antioxidative status of children with intraocular tumors (primary retinoblastoma) compared with children without tumors. The difference was not significant in adults. These enzymes may be of value in the early diagnosis of malignant intraocular tumor, especially retinoblastoma. PMID- 17416960 TI - Effect of pterygium excision by limbal conjunctival auotografting on tear function tests. AB - We investigated the effects of pterygium excision using the limbal conjunctival autografting technique on the tear function tests in 14 eyes of 13 patients. Since no difference was found in the Schirmer and tear break-up time tests at one and six months postoperative vs preoperative pterygium excision does not affect tear function test results during short-term follow up. PMID- 17416961 TI - Photodynamic therapy and intravitreal triamcinolone for extrafoveal choroidal neovascularization in neovascular age-related macular degeneration. AB - We report a case of extrafoveal choroidal neovascularization (CNV) secondary to neovascular age-related macular degeneration treated with photodynamic therapy (PDT) and intravitreal triamcinolone. Nine months following PDT and intravitreal triamcinolone, no ophthalmoscopic or angiographic evidence of recurrent CNV in the left eye was found. The intraocular pressure (IOP) increased from 10 mmHg on presentation to 20 mmHg at 9 months. No IOP-lowering agents were required. The mild nuclear sclerosis remained unchanged. PMID- 17416962 TI - Combination of photodynamic therapy, intravitreal triamcinolone injection, and standard laser photocoagulation in radiation retinopathy: a case report. AB - We report the combination of various therapy options in a 58-year-old man diagnosed with bilateral radiation retinopathy and left radiation optic neuropathy. Four mg/0.1 mL triamcinolone acetonide was injected intravitreally in both eyes followed by bilateral photodynamic treatment 2 weeks after. Bilateral panretinal photocoagulation and right focal laser treatment for residual macular edema were performed 2 months later. At 18 months, visual acuity is 20/200 in the right eye and hand motions in the left eye with no signs of active retinopathy. PMID- 17416963 TI - Blunt ocular trauma in one eye: a photo documentation. AB - Blunt ocular trauma can cause both structural and functional damage to the eye. We present a 13-year-old male who suffered a blunt injury with a wooden stick. Ocular examination revealed all the classic seven rings of trauma. Although ocular examination should always focus on these signs, all the signs are rarely present in a patient presenting with blunt trauma. Despite multistructural damage, the patient regained useful vision with medical management alone. PMID- 17416964 TI - Fungal endophthalmitis complicating subconjunctival injection of triamcinolone acetonide in anterior scleritis. AB - Fungal endophthalmitis following subconjunctival triamcinolone acetonide in anterior scleritis is a rare complication. Two patients with anterior scleritis, who received subconjunctival injection of triamcinolone developed fungal endophthalmitis. Both patients were positive by smear and polymerase chain reaction for 28S ribosomal RNA gene for fungus. Colletotrichum dematium was identified in one patient. Both patients responded well to pars plana vitrectomy and antifungal therapy. PMID- 17416965 TI - Gi and RGS proteins provide biochemical control of androgen receptor nuclear exclusion. AB - Nuclear localization of androgen receptors (ARs) is essential for their activity. Melatonin induces AR nuclear exclusion via increase in cGMP, calcium, and protein kinase C (PKC) activation, presumably through G-protein(s). The effects of regulators of G-protein signaling (RGS) on AR localization were studied in AR expressing PC3 cells. Gi-specific RGS10 inhibited melatonin but not cGMP-induced AR nuclear exclusion, independent of androgen. No evidence for Gq activation by melatonin was found. However, Gi/Gq-selective RGS4 inhibited AR nuclear exclusion downstream of PKC activation--an effect that was abrogated by constitutively active Gq. RGS10 and RGS4, but not RGS2, ablated the inhibitory effects of melatonin on AR reporter gene activity. For the first time, these data show regulation by Gi and Gi-specific RGS protein-mediated AR nuclear exclusion, which is potentially important in the treatment of AR-dependent cancers and neurodegenerative disorders. They also reveal a role for a Gq protein downstream of PKC activation in AR nuclear localization. PMID- 17416966 TI - Spinal and supraspinal changes in tumor necrosis factor-alpha expression following excitotoxic spinal cord injury. AB - The role of tumor necrosis factor-alpha (TNF-alpha) after spinal cord injury (SCI) is well characterized in the cord, but the impact of this inflammatory process on supraspinal levels is unknown. This study examines TNF-alpha mRNA and protein levels in the brains and spinal cords of mice after SCI. Mice received intraspinal injections of quisqualic acid (QUIS) to create an excitotoxic injury that is known to result in pain behaviors. An ELISA determined serum levels of TNF-alpha, whereas real-time PCR and Western blot analysis were used to determine mRNA and protein levels, respectively, at 3, 6, 12, 24, 48, 72 h, or 14 d postinjury. No difference existed in serum TNF-alpha levels between sham- and QUIS-injected animals. TNF-alpha mRNA in the cord was increased at 3, 6, 12, and 24 h in QUIS-injected animals relative to shams. TNF-alpha protein was elevated at 12 and 48 h postinjury. TNF-alpha mRNA levels in the brain were elevated at 12 and 24 h, with elevated protein levels at 6 h. Animals that developed pain behaviors had increased levels of TNF-alpha mRNA in the brain. Excitotoxic SCI results in altered TNF-alpha mRNA and protein levels in the cords and brains of mice within 6 h of injury. These changes likely contribute to the pathogenesis of injury within the cord. The role of TNF-alpha in the brain postinjury has not been defined but might contribute to the development of pain post-SCI. PMID- 17416967 TI - p35/Cyclin-dependent kinase 5 is required for protection against beta-amyloid induced cell death but not tau phosphorylation by ceramide. AB - Ceramide is a bioactive sphingolipid that can prevent calpain activation and beta amyloid (A beta) neurotoxicity in cortical neurons. Recent evidence supports A beta induction of a calpain-dependent cleavage of the cyclin-dependent kinase 5 (cdk5) regulatory protein p35 that contributes to tau hyperphosphorylation and neuronal death. Using cortical neurons isolated from wild-type and p35 knockout mice, we investigated whether ceramide required p35/cdk5 to protect against A beta-induced cell death and tau phosphorylation. Ceramide inhibited A beta induced calpain activation and cdk5 activity in wild-type neurons and protected against neuronal death and tau hyperphosphorylation. Interestingly, A beta also increased cdk5 activity in p35-/- neurons, suggesting that the alternate cdk5 regulatory protein, p39, might mediate this effect. In p35 null neurons, ceramide blocked A beta-induced calpain activation but did not inhibit cdk5 activity or cell death. However, ceramide blocked tau hyperphosphorylation potentially via inhibition of glycogen synthase kinase-3beta. These data suggest that ceramide can regulate A beta cell toxicity in a p35/cdk5-dependent manner. PMID- 17416968 TI - Testicular regulation of neuronal glucose and monocarboxylate transporter gene expression profiles in CNS metabolic sensing sites during acute and recurrent insulin-induced hypoglycemia. AB - Recurrent insulin-induced hypoglycemia (RIIH) impairs glucose counter-regulatory function in male humans and rodents and, in the latter, diminishes neuronal activation in CNS structures that monitor metabolic homeostasis, including the lateral hypothalamic area (LHA) and dorsal vagal complex (DVC). We investigated whether habituated neuronal reactivity in CNS sensing sites to hypoglycemia is correlated with modified monocarboxylate and/or glucose uptake by using quantitative real-time RT-PCR to analyze neuronal monocarboxylate transporter (MCT2) and glucose transporter variant (GLUT and GLUT4) gene expression profiles in the microdissected LHA, ventromedial nucleus hypothalamus (VMH), and DVC after one or multiple insulin injections. Because orchidectomy (ORDX) maintains uniform glycemic responses to RIIH in male rats, we also examined whether regional gene response patterns are testes dependent. In the intact male rat DVC, MCT2, GLUT3, and GLUT4 gene expression was not altered by acute hypoglycemia but was enhanced by RIIH. MCT2 and GLUT3 mRNA levels in the ORDX rat DVC did not differ among groups, but GLUT4 transcripts were progressively increased by acute and recurrent hypoglycemia. Precedent hypoglycemia decreased or increased basal MCT2 and GLUT4 gene expression, respectively, in the intact rat LHA; LHA GLUT3 transcription was augmented by RIIH in intact rats only. Acute hypoglycemia suppressed MCT2, GLUT3, and GLUT4 gene expression in the intact rat VMH, a response that was abolished by RIIH. In ORDX rats, VMH gene transcript levels were unchanged in response to one dose of insulin but were selectively diminished during RIIH. These data demonstrate site-specific, testes-dependent effects of acute and recurrent hypoglycemia on neuronal metabolic substrate transporter gene expression in characterized rat brain metabolic sensing loci and emphasize the need to assess the impact of potential alterations in glucose and lactate uptake during RIIH on general and specialized, e.g., metabolic monitoring, functions of neurons in those sites. PMID- 17416969 TI - Distribution of histone deacetylases 1-11 in the rat brain. AB - Although protein phosphorylation has been characterized more extensively, modulation of the acetylation state of signaling molecules is now being recognized as a key means of signal transduction. The enzymes responsible for mediating these changes include histone acetyl transferases and histone deacetylases (HDACs). Members of the HDAC family of enzymes have been identified as potential therapeutic targets for diseases ranging from cancer to ischemia and neurodegeneration. We initiated a project to conduct comprehensive gene expression mapping of the 11 HDAC isoforms (HDAC1-11) (classes I, II, and IV) throughout the rat brain using high-resolution in situ hybridization (ISH) and imaging technology. Internal and external data bases were employed to identify the appropriate rat sequence information for probe selection. In addition, immunohistochemistry was performed on these samples to separately examine HDAC expression in neurons, astrocytes, oligodendrocytes, and endothelial cells in the CNS. This double-labeling approach enabled the identification of specific cell types in which the individual HDACs were expressed. The signals obtained by ISH were compared to radiolabeled standards and thereby enabled semiquantitative analysis of individual HDAC isoforms and defined relative levels of gene expression in >50 brain regions. This project produced an extensive atlas of 11 HDAC isoforms throughout the rat brain, including cell type localization, providing a valuable resource for examining the roles of specific HDACs in the brain and the development of future modulators of HDAC activity. PMID- 17416970 TI - Localizations of endogenous APP/APP-proteolytic products are consistent with microtubular transport. AB - Dementia of the Alzheimer type (DAT) is associated with the accumulation of beta amyloid (A beta) peptides derived from beta-amyloid precursor protein (APP). Goldstein and coworkers have suggested that APP acts as a cargo receptor connecting post-Golgi vesicles and motor proteins. Sisodia and colleagues have suggested that APP is a passive passenger within the vesicles. Both views predict that one should be able to visualize colocalizations of APP with microtubules, the object of the present investigation. To avoid possible artifacts created by APP overexpression, we studied endogenous expression in a human neuroblastoma cell line (SK-N-SH). Using high resolution fluorescence microscopy and antibodies specific for the amino termini of APP and A beta sequences, we found that endogenous APP and A beta peptide immunoreactivities colocalized with microtubules in interphase cells. Disruption of microtubules, followed by fixation at various time points during repolymerization, allowed us to observe the sequence and timing of these colocalizations in interphase cells. In addition, to our surprise, we found that A beta immunoreactivities colocalize with the mitotic spindle, a bundle of specialized microtubules. Because of the condensed cytoplasm found in neurons, we suggest that SK-N-SH cells might be a more convenient experimental system for exploring the mechanisms that underlie these protein localizations and the pathology that might result from altered APP protein structure and function. PMID- 17416971 TI - PreproThyrotropin-releasing hormone 178-199 affects tyrosine hydroxylase biosynthesis in hypothalamic neurons: a possible role for pituitary prolactin regulation. AB - ProThyrotropin-releasing hormone (proTRH) is a prohormone widely distributed in many areas of the brain. After biosynthesis, proTRH is subjected to post translational processing to generate TRH and seven non-TRH peptides. Among these non-TRH sequences, we found previously that preproTRH178-199 could regulate the secretion of prolactin in suckled rats by their pups. Dopamine (DA), the main regulator of prolactin secretion, is produced in dopaminergic tyrosine hydroxylase (TH)-positive neurons in the hypothalamic arcuate nucleus (ARC). In this study we investigated whether prolactin release during the estrous sexual cycle is regulated by preproTRH178-199 through its effect on DA neurons of the ARC. We observed that biotinylated preproTRH178-199 bound to neurons in the ARC; this was higher during proestrus than during diestrus. Binding of preproTRH178 199 to DA neurons was seen only during proestrus in the ARC. Using primary neuronal hypothalamic cultures we found that preproTRH178-199 peptide decreased TH levels in a dose-responsive manner, whereas intra-ARC administration of preproTRH178-199 induced a 20-fold increase in plasma prolactin levels. Together, these results suggest a potential role for preproTRH178-199 in regulating dopaminergic neurons involved in the inhibition of pituitary prolactin release. PMID- 17416972 TI - Thrombin inhibits aquaporin 4 expression through protein kinase C-dependent pathway in cultured astrocytes. AB - Aquaporin 4 (AQP4) is a key molecule for maintaining water balance in the central nervous system, and its dysfunction might cause brain edema. However, little is known about the regulation of AQP4 expression. Because thrombin has been implicated in brain edema formation, the purpose of this study is to determine whether thrombin affects expression of AQP4 in astrocytes. Here, the effect of thrombin on AQP4 expression in vitro was evaluated using Western blot analysis and RT-PCR. Meanwhile, we investigated whether the effect of thrombin on AQP4 expression was due to protease-activated receptor 1 (PAR-1). In addition, we examined the role of protein kinase C (PKC) in the effect of thrombin on AQP4 expression using Western blot analysis. We found that thrombin did not affect cell viability at concentrations of 0.05, 0.5, 5, or 50 nM but killed astrocytes at concentrations of 500 nM, with approx 72% of astrocytes surviving at 500 nM thrombin. Our data showed that AQP4 protein expression achieved only 28% of controls in 500 nM thrombin treatment, even if astrocytes survived approx 72% of controls at 500 nM thrombin. Thrombin significantly inhibited AQP4 in a time- and dose dependent manner in vitro (p<0.05). Cathepsin-G, a thrombin PAR-1 inhibitor, reversed significantly (p<0.05) the effect of thrombin on AQP4 mRNA and protein expression in astrocytes. We also observed that PKC inhibitor H-7 or prolonged pretreatment with TPA can rapidly increase AQP4 expression (p<0.05). Thrombin might inhibit AQP4 expression in rat astrocytes, and this effect is possibly mediated by the PKC pathway. PMID- 17416973 TI - Biochemical properties of genetic recombinant xylanase II. AB - The aim of this study was to overexpress the xylanase II gene of Trichoderma reesei in Escherichia coli and determine the characteristics of the recombinant enzyme. Recombinant xylanase II gene was constructed by ligating the cDNA of xylanase, obtained from reverse transcriptase-polymerase chain reaction, and fused with NusA protein of pET-431b plasmid. An Ni2+-NTA affinity column was used to further purify the recombinant xylanase II. The molecular mass of the recombinant enzyme measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approx 76 kDa (including 55 kDa of NusA and 21 kDa of xylanase II), and the isoelectric point and specific activity were 7.5 and 225 U/mg, respectively. The optimal reaction temperature and pH for the recombinant enzyme were 50 degrees C and 4.0, respectively. The recombinant enzyme was stable at a pH range of 5.0-10.0 and maintained 95% residual activity after incubating at 30-35 degrees C for 30 min. The kinetic parameters KM and Vmax of the recombinant xylanase II were 13.8 mg/mL and 336 micromol/(mg.min), respectively, using birchwood xylan as the substrate. PMID- 17416974 TI - Fluorophore-assisted carbohydrate electrophoresis as detection method for carbohydrate-protein interactions. AB - Fluorophore-assisted carbohydrate electrophoresis (FACE) is a straightforward, sensitive method for determining the presence and relative abundance of individual (oligo)saccharide in a(n) (oligo)saccharide mixture. The single terminal aldehydes of (oligo)saccharides were tagged with the charged fluorophore 8-aminonaphthalene-1,3,6-trisulfonate (ANTS), and separated with high resolution on the basis of size by polyacrylamide gel electrophoresis. ANTS fluorescence labeling is not biased by (oligo)saccharide length. Therefore, band fluorescence intensity is directly related to the relative abundance of individual (oligo)saccharide moieties in heterogeneous sample. In the same time, it also indicates that FACE can be used to investigate the interactions of carbohydrates and proteins. PMID- 17416975 TI - Kinetics of gas-phase hydrolysis of ethyl acetate catalyzed by immobilized lipase. AB - Reactions catalyzed by supported enzymes present important advantages when compared with those in aqueous media or organic solvents: separation of enzymes from substrate is easily accomplished, enzyme stability may be improved, and control of the reaction products is more accurate. We present the experimental results of the kinetic study of ethyl acetate hydrolysis in gaseous phase catalyzed by a commercial immobilized lipase (Lipozyme IM; Novo Nordisk). The hydrolysis reaction was studied as a function of ethyl ester and water partial pressure at a constant temperature of 318 K. The amount of biocatalyst used was varied between 100 and 300 mg, and the reaction was studied in a flow-through glass microreactor. Under the conditions used, water was an important parameter in the gas-phase reaction. Activation energy was 24.8 kJ/mol and the overall order of reaction was one. Finally, a Bi-Bi reaction mechanism is proposed. PMID- 17416976 TI - Degradation of bisphenol A by Bacillus pumilus isolated from kimchi, a traditionally fermented food. AB - Novel bisphenol A (BPA)-degrading bacterial strains, designated as BP-2CK, BP 21DK, and BP-22DK, were isolated from kimchi, a traditionally fermented food. These isolates were identified as Bacillus pumilus and efficiently degraded BPA in a medium supplemented with nutrients such as peptone, beef extract, and yeast extract. Strains BP-2CK, BP-21DK, and BP-22DK successfully degraded 25, 25, and 50 ppm of BPA, respectively, and all strains exhibited BPA-degrading activity in the presence of 10% NaCl. Accumulation of the metabolites including 4 hydroxyacetophenone, one of the intermediates produced by the other BPA-degrading bacteria, was not observed in BPA degradation by the isolated strains. These results indicate that the isolated food-derived bacteria are applicable for the construction of efficient and safer systems for the removal of BPA. PMID- 17416977 TI - Decolorization of textile dye effluent by genetically improved bacterial strains. AB - Synthetic dyes are released into the environment from textile industrial effluents. The discharge of this colored wastewater into rivers and lakes leads to a reduction in sunlight penetration in natural water bodies, which, in turn, decreases both photosynthetic activity and dissolved oxygen concentration and is toxic to living beings. Bacterial isolates are optimized for growth and biomass production before using them for decolorizing dye effluent. The bacterial isolates Bacillus sp. 1 and Bacillus sp. 2 were employed at different percentages by volume with standard nutrient concentration. Of these bacterial isolates Bacillus sp. 2 recorded maximum color reduction. The pH and electrical conductivity (EC) were reduced in the decolorized effluent, and a reduction in biologic oxygen demand, chemical oxygen demand, total suspended solids, and total dissolved solids (TDS) were also observed. PMID- 17416978 TI - How polyamine synthesis inhibitors and cinnamic acid affect tropane alkaloid production. AB - Hairy roots of Brugmansia candida produce the tropane alkaloids scopolamine and hyoscyamine. In an attempt to divert the carbon flux from competing pathways and thus enhance productivity, the polyamine biosynthesis inhibitors cyclohexylamine (CHA) and methylglyoxal-bis-guanylhydrazone (MGBG) and the phenylalanine-ammonia lyase inhibitor cinnamic acid were used. CHA decreased the specific productivity of both alkaloids but increased significantly the release of scopolamine (approx 500%) when it was added in the mid-exponential phase. However, when CHA was added for only 48 h during the exponential phase, the specific productivity of both alkaloids increased (approx 200%), favoring scopolamine. Treatment with MGBG was detrimental to growth but promoted release into the medium of both alkaloids. However, when it was added for 48 h during the exponential phase, MGBG increased the specific productivity (approx 200%) and release (250- 1800%) of both alkaloids. Cinnamic acid alone also favored release but not specific productivity. When a combination of CHA or MGBG with cinnamic acid was used, the results obtained were approximately the same as with each polyamine biosynthesis inhibitor alone, although to a lesser extent. Regarding root morphology, CHA inhibited growth of primary roots and ramification. However, it had a positive effect on elongation of lateral roots. PMID- 17416979 TI - Preparation of chitosan nanoparticles as carrier for immobilized enzyme. AB - This work investigated the preparation of chitosan nanoparticles used as carriers for immobilized enzyme. The morphologic characterization of chitosan nanoparticles was evaluated by scanning electron microscope. The various preparation methods of chitosan nanoparticles were discussed and chosen. The effect of factors such as molecular weight of chitosan, chitosan concentration, TPP concentration, and solution pH on the size of chitosan nanoparticles was studied. Based on these results, response surface methodology was employed. The results showed that solution pH, TPP concentration, and chitosan concentration significantly affected the size of chitosan nanoparticles. The adequacy of the predictive model equation for predicting the magnitude orders of the size of chitosan nanoparticles was verified effectively by the validation data. Immobilization conditions were investigated as well. The minimum particles size was about 42 +/- 5 nm under the optimized conditions. The optimal conditions of immobilization were as follows: one milligram of neutral proteinase was immobilized on chitosan nanoparticles for about 15 min at 40 degrees C. Under the optimized conditions, the enzyme activity yield was 84.3%. PMID- 17416980 TI - Modeling and computational simulation of dilution and biochemical materials balance equations for partially emptied batch reactors. AB - Sequencing batch reactors (SBRs) including aerobic SBRs and anaerobic SBRs (ASBRs) are partially emptied batch reactors that are widely used as bioprocesses in pollution control. We present dilution and biochemical materials balance modeling equations and simulation results for the partially emptied batch reactors, especially for ASBR treatment of low-strength wastewater. The simulated substrate and microbial concentrations for both dilution and materials balance equations follow the same pattern during both feeding and reaction times. However, the results of the materials balance equations show microbial activities during feeding as well as during reaction times and were found to be more appropriate for the biologic system in which substrate removal is associated with microbial growth. Furthermore, the simulation results point to the need to foster high microbial accumulation in the system during startup to optimize the process performance and the need to operate the system at a short reaction time, especially for low substrate concentrations. The results were found to be in agreement with the results of prior laboratory studies. PMID- 17416981 TI - Characterization of immobilized nuclease P1. AB - To improve the efficiency of the use of nuclease P1, enzyme immobilization technology was applied using nuclease P1. Characterization of immobilized nuclease P1 on different supports was studied. The results showed that the optimum pH and temperature of nuclease P1 immobilized via different supports were enhanced. The immobilized enzyme was obviously stable when stored for long periods and was reusable. The best results were obtained when nuclease P1 was immobilized on chitosan nanoparticles. The nanoparticles were applied to protect the activity of nuclease P1 and improved enzyme activity by 13.17% over that of free nuclease P1 at the same conditions. The Michaelis constant Km and Vmax were determined for free and immobilized enzyme as well. PMID- 17416982 TI - Expression of Cu, Zn-superoxide dismutase gene from Saccharomyces cerevisiae in Pichia pastoris and its resistance to oxidative stress. AB - The gene for the Cu, Zn-superoxide dismutase (SOD) from the yeast Saccharomyces cerevisiae was cloned, characterized, and overexpressed in the methylotrophic Pichia pastoris. The sod gene sequence obtained is 465 bp and encodes 154 amino acid residues. The sod gene sequence was cloned into the pPIC9K vector, yielding pAB22. The linearized pAB22 DNA, digested with restriction enzyme SacI, was transformed into the genome of the GS115 strain of the yeast P. pastoris. The SOD was purified from the cultured yeast by ammonium sulfate precipitation and DEAE cellulose column chromatography. This relatively simple purification method produced a single band on analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The overexpressed SOD protein was shown to have immunologically biologic activity and to be enzymatically active. The yeast overexpressing Cu, Zn SOD appeared to be more resistant to oxidative stress such as paraquat, menadione, and heat shock. PMID- 17416983 TI - Manipulation of cell-cell adhesion using bowtie-shaped microwells. AB - Traditional methods to study cell-cell adhesion have been limited by their inability to manipulate cell-cell interactions without simultaneously affecting other microenvironmental factors. Here we describe a novel method that enables the culture of cells with precise simultaneous control of both cell-cell and cell substratum adhesion. Using microfabricated stamps of poly(dimethylsiloxane), we construct bowtie-shaped agarose microwells into which cells can be cultured. The degree to which cells spread is controlled by the size of the microwell; cell cell contacts form between neighboring cells within the microwell. This chapter describes the details of stamp fabrication, agarose microwell construction, and cell culture in micropatterned substrata. PMID- 17416984 TI - Separation of cell-cell adhesion complexes by differential centrifugation. AB - The number of proteins found associated with cell-cell adhesion substructures is growing rapidly. Based on potential protein-protein interactions, complex protein networks at cell-cell contacts can be modeled. Traditional studies to examine protein-protein interactions include co-immunoprecipitation or pull-down experiments of tagged proteins. These studies provide valuable information that proteins can associate directly or indirectly through other proteins in a complex. However, they do not clarify if a given protein is part of other protein complexes or inform about the specificity of those interactions in the context of adhesion substructures. Thus, it is not clear if models compiled from these types of studies reflect the combination of protein interactions in the adhesion complex in vivo for a specific cell type. Therefore, we present here a method to separate cell-cell contact membrane substructures with their associated protein complexes based on their buoyant behavior in iodixanol density gradients. Analysis of 16 proteins of the apical junctional complex (AJC) in epithelial Madin-Darby canine kidney cells revealed a more simple organization of the AJC adhesion complex than that predicted from the combination of all possible protein protein interactions defined from co-immunoprecipitation and pull-down experiments. PMID- 17416985 TI - Analysis of neutrophil chemotaxis. AB - Neutrophils are the initial responders to bacterial infection or other inflammatory stimuli and comprise a key component of the innate immune response. In addition to their unique morphology and antimicrobial activity, neutrophils are characterized by the ability to migrate rapidly up shallow gradients of attractants in vivo. The directed migration of neutrophils, referred to as chemotaxis, requires the temporal and spatial regulation of intracellular signaling pathways allowing the neutrophil to detect a gradient of attractant, polarize, and migrate rapidly toward the highest concentration of the chemoattractant. A challenge to understanding neutrophil chemotaxis is the inherent difficulty encountered when working with primary neutrophils, which are difficult to purify in the resting state, are not easily transfected, are terminally differentiated, and have a short life span after purification. Here we discuss neutrophil purification methods and chemotaxis assays and provide methodology for working with a neutrophil-like cell line, the HL-60 promyelocytic leukemia cell line. We also discuss methods for HL-60 transfection using retroviral approaches and chemotaxis assays used with differentiated HL-60 cells. PMID- 17416986 TI - Analysis of leukocyte migration through monolayers of cultured endothelial cells. AB - In this chapter methods are described for analyzing the adhesion and migration of isolated leukocytes on endothelial cell monolayers that have been cultured on different substrates and treated with cytokines. When endothelial cells are grown on porous filters inserted in wells, the levels of leukocyte adhesion and migration are calculated from the number added and the numbers retrieved from the upper and lower chambers. Fluorescence microscopic examination of the fixed filters can be used to ascertain whether leukocytes are retained above or below the filter. Direct observations of the time course of migration can be made when endothelial cells are cultured in six-well plates after leukocytes are allowed to settle onto them for a short period. In a more specialized assay, leukocytes are perfused through glass capillaries coated with endothelial cells, and again, direct video-microscopic observations are made. In this assay all stages of capture, immobilization, and migration can be followed. In general, the filter based assay has the highest throughput and greatest ease of use but yields less detailed information, whereas the flow-based assay is most difficult to set up but is most physiologically relevant. PMID- 17416987 TI - Biochemical purification of pseudopodia from migratory cells. AB - Cell migration requires the formation of a leading pseudopodium (lamellipodium) in the direction of movement. This process requires signal amplification to facilitate directional sensing mechanisms that lead to actin-mediated membrane extension. However, it has been difficult to study pseudopodia formation because it has not been possible to purify this structure for biochemical analysis. Here we describe a method to biochemically purify the protruding pseudopodium from the cell body compartment using polycarbonate microporous filters. Cells are cultured on top of 3.0-microm porous filters and allowed to extend pseudopodia through the small pores to the undersurface in response to a gradient of either chemokine or extracellular matrix (ECM) protein. Pseudopodia and cell bodies are then differentially scraped from the filter surface into lysis buffer for biochemical analysis. Using this method, it is possible to identify novel pseudopodium and cell body proteins as well as study the spatiotemporal organization of signaling processes that regulate pseudopodium formation and cell polarity. This method will help facilitate our understanding of how cells protrude pseudopodia through small openings in the ECM and vasculature during cancer cell invasion, immune cell surveillance, and embryonic development. PMID- 17416988 TI - Dynamic assessment of cell-matrix mechanical interactions in three-dimensional culture. AB - Cell-matrix mechanical interactions play a defining role in a range of biological processes such as developmental morphogenesis and wound healing. Despite current agreement that fibroblasts exert mechanical forces on the extracellular matrix (ECM) to promote structural organization of the collagen architecture, the underlying mechanisms of force generation and transduction to the ECM are not completely understood. Investigation of these processes has been limited, in part, by the technical challenges associated with simultaneous imaging of cell activity and fibrillar collagen organization. To overcome these limitations, we have developed an experimental model in which cells expressing proteins tagged with enhanced green fluorescent protein are plated inside fibrillar collagen matrices, and high magnification time-lapse differential interference contrast and fluorescent imaging is then performed. Using this system, focal adhesion movement and reorganization in isolated cells can be directly correlated with collagen matrix deformation and changes in the mechanical behavior of fibroblasts can be assessed over time. PMID- 17416989 TI - Quantitative analyses of cell adhesion strength. AB - Biochemical and mechanical analyses of integrin-mediated cell adhesion have been limited by the inability to apply controlled detachment forces and the inherent complexities of the adhesive process, including cell spreading, integrin clustering, cytoskeletal interactions, and non-uniformly distributed focal complexes. A comprehensive set of techniques to analyze mechanical and biochemical events at the cell-extracellular matrix interface is presented. The spinning disk assay provides a rigorous hydrodynamic assay to measure adhesion strength. Crosslinking/extraction and wet-cleaving biochemical approaches isolate and quantify integrins bound to their ligands and adhesive components recruited to focal adhesions. These techniques provide an experimental framework for the rigorous analysis of cell adhesion. PMID- 17416990 TI - Using RNA interference to knock down the adhesion protein TES. AB - RNA interference (RNAi) is a specific and efficient method to knock down protein levels using small interfering RNAs (siRNAs), which target mRNA degradation. RNAi can be used in mammalian cell culture systems to target any protein of interest, and several studies have used this method to knock down adhesion proteins. We used siRNAs to knock down the levels of TES, a focal adhesion protein, in HeLa cells. We demonstrated knockdown of both TES mRNA and TES protein. Although total knockdown of TES was not achieved, the observed reduction in TES protein was sufficient to result in a cellular phenotype of reduced actin stress fibers. PMID- 17416991 TI - Assaying calpain activity. AB - The calpains represent a well-conserved family of intracellular proteases that exhibit broad substrate specificity and consequently influence many cellular processes. Calpain activity regulates the turnover of integrin-linked focal adhesions, which controls cell adhesion to extracellular matrix substrates, cell migration across such substrates, as well as the signalling output from the associated integrin receptors. Thus, calpain activity regulates both the physical interaction and biochemical communication between cells and the extracellular environment that is vital for normal cell function. Modulation of calpain activity is associated with a number of human diseases, and recent studies have identified calpain activity as an important therapeutic target in several disease areas. These studies have driven the development of in vitro and live-cell-based assays for high-throughput identification and evaluation of calpain inhibitors. However, many of the unique features of calpain activity devalue the physiological relevance of existing assays. In this chapter we describe a modified approach that monitors calpain activity by taking into account several of the factors that control calpain activity and substrate specificity under physiological conditions. The development of such second-generation assays may help to identify more effective calpain intervention strategies. PMID- 17416992 TI - Analysis of focal adhesions and cytoskeleton by custom microarray. AB - Focal adhesions and the cell cytoskeleton (intermediate filaments, microfilaments, microtubules) are involved in mechanotransduction-both direct (transduction of mechanical forces to the nucleus) and indirect (transduction of chemical signaling cascades to the nucleus). Thus, observation of changes in focal adhesion and cytoskeletal organization can be invaluable in research such as drug treatments and medical material testing in vitro. Here we describe how to stain human fibroblasts for vinculin (located to focal adhesions), actin (microfilaments), tubulin (microtubules), and vimentin (intermediate filaments) and how to perform custom microarray experiments. Comparative analysis of the immunofluorescence and array data should allow the researcher to build up a global picture of changes to both direct and indirect mechanotransduction through the cytoskeleton from focal adhesions. PMID- 17416993 TI - Proteomic analysis of cell surface membrane proteins in leukemic cells. AB - Plasma membrane proteins play a key role in cellular processes such as migration, adhesion, and cell survival. The comprehensive annotation of the leukemic cell plasma membrane proteome allows the identification of proteins that may be involved in the pathogenesis of disease and may provide novel therapeutic targets. The identification of known adhesion molecules or novel proteins with similar attributes to adhesion molecules provides the starting point for the generation of hypothesis on the role of these proteins in adhesion processes. In order to identify these novel proteins, we have developed a proteomics methodology using purified plasma membranes prepared from human leukemic cells. PMID- 17416994 TI - Bioinformatic analysis of adhesion proteins. AB - Proteins that mediate cell-cell and cell-extracellular matrix (ECM) adhesion have been fundamental in the evolution of multicellular animals. Fibrillar collagens, proteoglycans, integrins, and cadherins are present in all animals from sponges to mammals, and many other adhesion proteins have arisen during animal evolution. In general, adhesion proteins are large multidomain molecules and are encoded in larger gene families in vertebrates than in invertebrates. With the increasing availability of completely sequenced genomes representing different points on the animal tree of life, bioinformatics is proving to be a very valuable approach for the analysis of the domain organization and relationships of adhesion proteins, which can direct or enhance experimental tests. Here we describe, with examples from the literature, the major methods for identifying sequence homologies; analyzing domain organization and potential for oligomerization; analyzing sequence relationships by multiple sequence alignments and phylogenetic trees, and assessing adhesion proteins as components of functional pathways and tissue systems through comparative genomics. PMID- 17416995 TI - Analysis of integrin dynamics by fluorescence recovery after photobleaching. AB - Cell migration is a complex cellular behavior that involves the controlled reorganization of the link between the actin cytoskeleton and the extracellular matrix. This mechanical connection is provided by transmembrane receptors of the integrin family. Integrins are heterodimeric receptors that undergo an allosteric switch when activated by external or intracellular signals, providing binding sites for ligands of the extracellular matrix and actin-associated cytoplasmic adapter proteins. Techniques such as fluorescence recovery after photobleaching (FRAP) are used to analyze the remodeling of green fluorescent protein (GFP) tagged integrin receptors within focal adhesions, demonstrating that the dynamics of the remodeling of integrins in substrate adhesion sites is carefully regulated by extracellular ligands, cytoskeletal adapter proteins, and the actin cytoskeleton. FRAP analysis of GFP-tagged integrins is a tool that allows one to detect and dissect the internal dynamics of apparently immobile focal adhesions, allowing one to perceive the hierarchies and mechanisms of focal adhesion formation and dispersion. PMID- 17416996 TI - Double-hydrogel substrate as a model system for three-dimensional cell culture. AB - When cultured on two-dimensional surfaces, most adherent cells show profound differences from those in their native habitats. In addition to chemical factors, it is likely that both physical parameters, such as substrate rigidity, and topographical factors, such as the asymmetry in integrin anchorage, play a major role in the differences. We have designed a simple culture system that provides flexible, adhesive substrates for both dorsal and ventral cell surfaces. Fibroblasts in this system show the spindle or stellate morphology found in native tissues. The ease of preparation, versatility, and optical quality of this model system should greatly facilitate the understanding of cellular behavior and functions in vivo. PMID- 17416997 TI - In vitro actin assembly assays and purification from Acanthamoeba. AB - The actin cytoskeleton is essential to all eukaryotic cells. In addition to playing important structural roles, assembly of actin into filaments powers diverse cellular processes, including cell motility and endocytosis. Actin polymerization is tightly regulated by various cofactors, which control spatial and temporal assembly of actin as well as the physical properties of these filaments. Development of an in vitro model of actin polymerization from purified components has allowed for great advances in determining the effects of these proteins on the actin cytoskeleton. The pyrene actin assembly assay is a powerful tool for determining the effect of a protein on the kinetics of actin assembly, either directly or as mediated by proteins such as nucleators or capping factors. In addition, fluorescently labeled phalloidin can be used to visualize the filaments that are created in vitro to give insight into how these proteins influence actin filament superstructure. PMID- 17416998 TI - Light, photoreceptors, and circadian clocks. AB - Research over the past decade has focused increasingly on the photoreceptor mechanisms that regulate the circadian system in all forms of life. Some of the results to emerge are surprising. For example, the rods and cones within the mammalian eye are not required for the alignment (entrainment) of circadian rhythms to the dawn-dusk cycle. There exists a population of directly light sensitive ganglion cells within the eye that act as brightness detectors; these regulate both circadian rhythms and melatonin synthesis. An understanding of these "circadian photoreceptor" pathways, and the features of the light environment used for entrainment, have been and will continue to be heavily dependent on the appropriate use and measurement of light stimuli. Furthermore, if results from different laboratories, or species, are to be compared in any meaningful sense, standardized methods for light measurement and manipulation need to be adopted by circadian biologists. To this end, we describe light measurement in terms of both radiometric and photometric units and consider the appropriate use of light as a stimulus in circadian experiments. In addition, the construction of action spectra has been very helpful in associating photopigments with particular responses in a broad range of photobiological systems. Because the identity of the photopigments mediating circadian responses to light are often not known, we have also taken this opportunity to provide a step-by-step approach to conducting action spectra, including the construction of irradiance response curves, the calculation of relative spectral sensitivities, photopigment template fitting, and the underlying assumptions behind this approach. The aims of this chapter are to provide an accessible introduction to photobiological methods and explain why these approaches need to be applied to the study of circadian systems. PMID- 17416999 TI - Statistical analysis of biological rhythm data. AB - The author has developed an ensemble of digital signal analysis techniques applicable to biological time series containing circadian and ultradian periodicities that is of very high resolution and functions well even in the presence of extreme noise and trend. A method for quantifying the significance, strength, and regularity of the rhythmic process is included. To illustrate these techniques, the author presents analyses of artificial periodic data containing varying amounts of noise, trend, and multiple periodicities. The periods and amplitudes of circadian and, where included, ultradian periodicities, and all other components of the test signals are known exactly. Analyses are illustrated in a step-by-step manner and the results are compared with the known input parameters. Trends are removed; spectra, autocorrelation functions, and rhythmicity indices are produced and discussed. References covering theory and details of all analyses are supplied. All programs employed are available from the author free of charge. PMID- 17417000 TI - Rhythmic conidiation in Neurospora crassa. AB - In the filamentous fungus Neurospora crassa the production of asexual spores (conidia) is regulated by its circadian clock. When the fungus is grown on a thin layer of agar medium in long growth tubes (so-called "race tubes"), restricting its growth to one direction only, bright orange bands are clearly visible. This banding pattern persists with a periodicity of approx 24 h in the absence of any environmental stimuli. The bands are formed by alternating zones of nonsporulating mycelium and mycelium laden with orange conidia. Assaying Neurospora conidiation on race tubes is a simple yet powerful and versatile tool for studying the circadian clock of this model organism. PMID- 17417001 TI - Monitoring and analyzing Drosophila circadian locomotor activity. AB - In the 1970s, the intriguing discovery of autonomous circadian rhythmicity at the behavioral level in Drosophila set the starting point for one of the most remarkably rapid advancements in the understanding of the genetic and molecular bases of a complex behavioral trait. To this end, the design of appropriate electronic devices, apt to continuously monitor behavioral activity, has proven to be fundamental to such progress. In particular, most of the mutational screens performed to date in the search for genes involved in circadian rhythmicity were based on monitoring Drosophila mutants for alterations in the circadian pattern of locomotor activity. Many different experimental paradigms, based on the use of circadian locomotor activity monitors, have been developed. Experiments can be designed to determine (1) the natural period, (2) the capacity to adapt to day night cycles with photoperiods of differing length, and (3) the phase of the circadian activity cycles with respect to the entraining stimulus. Here we describe some of the rationale and the steps required to set up experiments to monitor circadian locomotor activity in Drosophila. Suggestions for the statistical analysis of the data obtained in such experiments are also provided. PMID- 17417002 TI - Automated video image analysis of larval zebrafish locomotor rhythms. AB - A method is described for measurement of the circadian activity rhythms of up to 150 larval zebrafish simultaneously with a single video image analysis system. Most of the required equipment and software are commercially available, although some components are custom-built. PMID- 17417003 TI - Locomotor activity in rodents. AB - Many of the behavioral parameters exhibited by an organism show daily fluctuations. These may persist under constant environmental conditions, demonstrating that they are governed by an endogenous (circadian) clock. The monitoring of locomotor activity in rodents is probably one of the most common methods to track this endogenous timing system. The analysis of locomotor activity rhythms can provide several parameters that may be used to describe the status of this endogenous clock. In the past few years several companies have developed hardware and software systems that allow the collection and the analysis of activity data using a personal computer. PMID- 17417004 TI - Analysis of circadian leaf movement rhythms in Arabidopsis thaliana. AB - Arabidopsis thaliana is the model organism for the study of the higher plant circadian clock. The physiological change in position of young leaves and cotyledons in Arabidopsis seedlings reveals an overt circadian rhythm. Measuring these leaf movements provides a simple and reliable assay of the plant circadian clock and, unlike systems based on the firefly luciferase reporter gene, requires no prior genetic manipulation of the plant. As such, leaf movement can be used to measure circadian rhythms in plants lacking luciferase reporter genes, or as an independent measure of the clock in plants that do possess the transgene. The imaging system described in this chapter can also be adapted to measure circadian rhythms in other plant species displaying rhythmic leaf movements. PMID- 17417005 TI - Detection of rhythmic bioluminescence from luciferase reporters in cyanobacteria. AB - The unicellular cyanobacterium Synechococcus elongatus PCC 7942 is the model organism for studying prokaryotic circadian rhythms. Although S. elongatus does not display an easily measurable overt circadian behavior, its gene expression is under circadian control; hence, a "behavior" is created by linking a cyanobacterial promoter to either the bacterial luxAB or firefly luc luciferase genes to create reporter fusions whose activity can be easily monitored by bioluminescence. Numerous vectors have been created in our lab for introducing luciferase reporter genes into the S. elongatus chromosome. A choice of methods and equipment to detect light production from the luciferase fusions provides a means for high-throughput, automated mutant screens as well as testing rhythms from two promoter fusions within the same cell culture. PMID- 17417006 TI - Analysis of rhythmic gene expression in adult Drosophila using the firefly luciferase reporter gene. AB - The study of circadian clock function in Drosophila relies heavily on the analysis of rhythmic gene expression. Typically, individuals or groups of flies collected during a specific time of the circadian day need to be sacrificed, followed by the extraction of clock gene products. This procedure makes it impossible to analyze molecular rhythms in an individual over time. To measure clock gene expression within the living animal, firefly luciferase can be used as real-time reporter gene. This chapter describes how rhythmic expression of clock or clock-controlled genes can be measured in living adult Drosophila. A survey of all existing clock-related luciferase transgenics is given. PMID- 17417007 TI - Monitoring circadian rhythms in Arabidopsis thaliana using luciferase reporter genes. AB - Arabidopsis thaliana is internationally adopted as the model plant species for molecular genetics. As such a huge range of resources are available for its study. Arabidopsis does not display any obvious circadian rhythms; however, rhythms in gene expression can be readily detected. For this reason the promoters of rhythmically expressed genes have been fused to the firefly luciferase gene. Using this reporter gene we have developed a number of automated techniques for monitoring luciferase activity in vivo in plants. This provides us with a robust and high-throughput assay for the circadian clock in Arabidopsis. PMID- 17417008 TI - Specialized techniques for site-directed mutagenesis in cyanobacteria. AB - Synechococcus elongatus PCC 7942 is an excellent model system for studying the molecular mechanism of the circadian clock in cyanobacteria. The "plastic" genetic characteristics of this organism have facilitated the development of various methods for mutagenesis of its chromosome. These methods are based on homologous recombination between the chromosome and foreign DNA, introduced to the cyanobacteria by either transformation or conjugation. Here we describe different approaches to mutagenize the chromosome of S. elongatus, including insertional mutagenesis, hit-and-run allele replacement, rps12-mediated gene replacement, and regulated expression of genes from ectopic sites, the neutral sites of the S. elongatus genome. PMID- 17417009 TI - Novel strategies for identification of clock genes in Neurospora with insertional mutagenesis. AB - As the molecular mechanism of circadian clocks has reached high complexity, the fungal model system, Neurospora crassa, is increasingly important for clock research. It offers the possibility of extensive biochemical experimentation and thorough description of circadian properties. Realization of the full potential is dependent on efficient, high-throughput methods. We have combined several protocols to develop abundant and inexpensive production of mutants, and subsequent identification of the affected gene. We applied a novel screening protocol and, after screening several hundred mutants, identified a known clock gene, frequency. Furthermore, the methods described here can easily be adapted to various insertional constructs (e.g., those with alternative selection markers or that facilitate overexpression) or combined with strains carrying clock-regulated reporter genes. PMID- 17417010 TI - Mutagenesis with Drosophila. AB - Forward genetics in Drosophila has profoundly affected our understanding of circadian rhythms in this organism and, more generally, in the animal kingdom. Most Drosophila pacemaker genes were discovered through the isolation of gene variants affecting the free-running period of the circadian pacemaker. There are different ways to mutagenize flies. An alkylating agent can be used to randomly alter the fly genome, or transposable elements can be mobilized to disrupt or increase the expression of the targeted genes. The advantages of these different methods are complementary. PMID- 17417011 TI - Mutagenesis in Arabidopsis. AB - Ethyl methane sulfonate (EMS) mutagenesis in Arabidopsis is the most widely used mutagenesis technique. EMS has high mutagenicity and low mortality and can be used in any laboratory with a fume hood. The chemical principle of EMS mutagenesis is simple; it is based on the ability of EMS to alkylate guanine bases, which results in base mispairing. An alkylated guanine will pair with a thymine base, resulting primarily in G/C to A/T transitions, which ultimately results in an amino acid change or deletion. There are several advantages to EMS mutagenesis compared with other mutagenesis techniques available for Arabidopsis. First, EMS generates a high density of nonbias irreversible mutations in the genome, which permits saturation mutagenesis without having to screen a large number of individual mutants. Second, EMS mutagenesis not only generates loss-of function mutants, but can also generate novel mutant phenotypes, which include dominant or gain-of-function versions of proteins owing to alterations of specific amino acids. This chapter describes the use of EMS mutagenesis in Arabidopsis and how mutagenized plant populations should be handled after the mutagenesis event. PMID- 17417012 TI - Yeast two-hybrid screening. AB - Yeast two-hybrid screening represents a sensitive in vivo method for the identification and analysis of protein-protein interactions. The principle is based on the ability of a separate DNA-binding domain (DNA-BD) and activation domain (AD) to reconstitute a functional transactivator when brought into proximity. In the MATCHMAKER yeast two-hybrid system, a bait protein is expressed as a fusion to the GAL4 DNA-BD, whereas the prey protein is expressed as a fusion to the GAL4 AD. When a bait and a prey protein interact, the DNA-BD and AD form a functional transactivator, resulting in activation of reporter gene expression in yeast reporter strains. The method described in this chapter can be used to identify novel protein interactions, analyze protein-protein interactions between two known proteins, as well as dissect interacting protein domains. PMID- 17417013 TI - Microarrays: quality control and hybridization protocol. AB - Microarray technology is an exciting and promising tool, and is increasingly employed for studying circadian rhythms. To obtain optimal results from this technology, it is important to perform quality control experiments before engaging in genome-wide expression analysis. In this chapter, we provide an overview of quantitative polymerase chain reaction experiments using the ABI PRISM 7900HT system for quality control of samples, and microarray experiments using the Affymetrix GeneChip system for genome-wide expression analysis. PMID- 17417014 TI - Microarrays: statistical methods for circadian rhythms. AB - Microarrays are promising tools that are increasingly being applied to the study of circadian rhythms. The large and complex datasets they generate, however, mean they require a new approach on how to design experiments, handle datasets, translate results, and derive conclusions. This technology also requires statistical methods for the correct interpretation of data generated by the microarrays. In this chapter, we provide an overview of analytical methods applied to microarray experiments for the identification of genes with circadian expression. PMID- 17417015 TI - Identification of clock genes using difference gel electrophoresis. AB - Proteomics is the study of the complete set of proteins encoded by the genome. The study of the proteome involves the investigation of changes in protein abundance, localization, involvement in multiprotein complexes, and detection of different protein isoforms and posttranslational modifications under defined conditions, such as the circadian cycle. This type of approach complements comparative gene expression studies providing additional information with respect to posttranscriptional processing. One of the key techniques used to study the proteome is two-dimensional gel electrophoresis. This technique has the ability to separate complex protein mixtures with high resolution. A significant improvement in this technology has been development of difference gel electrophoresis. Here, proteins are first labeled with one of three spectrally resolvable fluorescent cyanine dyes before being separated in two dimensions according to their charge and size, respectively. Multiplexing can accurately and reproducibly quantify protein expression across multiple gels. A multiple-gel approach allows the detection of differentially expressed protein spots using statistical methods to compare expression across different experimental groups. The proteins can be subsequently identified by mass spectrometric methods. This approach now allows more complex experimental designs, such as the time course experiments essential to the study of circadian rhythms. PMID- 17417016 TI - Isolation of total RNA from Neurospora mycelium. AB - In filamentous fungi, including the model organism Neurospora crassa, plentiful biological tissue from which RNA can be extracted may be obtained by allowing fungal spores to germinate and form a mycelium in liquid culture. The mycelium constitutes a mosaic of multinuclear, tubular filaments known as hyphae or mycelia. In general, when exposed to air, fungal hyphae quickly start to develop spores, which are often colorful. However, when submerged in liquid under rapid agitation large amounts of vegetatively growing mycelium can be obtained, which can be easily harvested by means of filtration. To preserve the physiological state of the culture, the mycelium is snap-frozen, and then to free its contents, the mycelium is ground under liquid nitrogen to break all hyphal structures. Here a method to extract high-quality total RNA from Neurospora mycelium using TRIzol reagent is described. PMID- 17417017 TI - RNA extraction from Drosophila heads. AB - In Drosophila, input, pacemaker, and output genes are expressed circadianly. mRNA oscillations contribute largely to these rhythms. Determining RNA levels of circadian genes is thus frequently necessary to understand their regulation, or the effect of mutations and genetic manipulations on the function of the circadian pacemaker. RNA extraction is the prelude to several techniques aimed at measuring RNA levels. The procedure presented in this chapter is a rapid method to obtain a clean preparation of total RNA from fly heads that can be used for RNase protection, Northern blots, and real-time polymerase chain reaction (see Chapters 23-25). PMID- 17417018 TI - Extraction of plant RNA. AB - This protocol details an RNA preparation for medium-scale, high-purity RNA production from higher plants. It uses hot acid phenol with standard sodium acetate ethanol precipitation and is suitable for producing RNA for both Northern blotting and enzyme-based downstream applications such as RT-PCR and microarray studies. PMID- 17417019 TI - RNA extraction from mammalian tissues. AB - Purification of intact RNA is the primary step of many molecular biology techniques, including Northern blotting, RNase protection, quantitative polymerase chain reaction, and microarray assays. RNA extraction is typically conducted using either a phenol-chloroform or a solid phase method. This article concentrates primarily on the former approach, which is highly versatile, and is easily adapted to different tissues ranging from whole organs down to submillimeter biopsy punches. The major problem with RNA extraction is the ubiquitous nature of RNases, enzymes that rapidly degrade RNA. RNases may be difficult to eliminate, although with care and appropriate countermeasures, reproducible extraction of high-quality RNA from important biological samples should be attainable. This article focuses on the isolation of RNA from the tissue collection step, homogenization all the way through to the quantification of the purified nucleic acid, providing guidelines for the prevention of the problems associated with RNAse contamination. PMID- 17417020 TI - Northern analysis of sense and antisense frequency RNA in Neurospora crassa. AB - In Northern analysis the presence of specific RNA transcripts is detected and their quantity can be estimated. RNA is separated using denaturing agarose gel electrophoresis and is subsequently transferred and fixed to a solid support, such as a nitrocellulose filter. When labeled probes are hybridized to these immobilized RNA molecules, their presence can be visualized by autoradiography. Here we describe Northern hybridization using radioactively labeled riboprobes to show circadian expression of endogenous sense and antisense frequency RNA in the filamentous fungus Neurospora crassa. PMID- 17417021 TI - RNase protection assay. AB - The RNase protection assay is a standard approach to determine mRNA levels of a gene of interest in different tissues, developmental stages, or times of the day. Splicing or promoter variants can be studied with specific probes. It is widely used in chronobiology to study the temporal profile of expression of circadian genes and the effects of genetic manipulation on these oscillations. Methods to generate the riboprobes and to perform the RNase protection assay itself are described in this chapter. PMID- 17417022 TI - Quantitative polymerase chain reaction. AB - Quantitative PCR (qPCR) has entered widespread use with the increasing availability of real-time PCR. By the incorporation of fluorescent dyes in the reaction mixture, increases in amplification products can be monitored throughout the reaction, enabling measurements to be taken in the exponential phase of the reaction, before the reaction plateau. Whatever the platform or chemistry involved, the starting point of a real-time assay is a tissue-specific RNA and the end point of a real-time reaction is an amplification plot. As such, rather than focusing on specific platforms or chemistries, herein we address the basic principles that underlie sample preparation, experimental design, use of internal controls, assay considerations, and approaches to data analysis. The advent of real-time PCR has enabled high-throughput analysis of multiple transcripts from small tissue samples, with an unparalleled dynamic range and sensitivity. However, to new users, this technique may seem to require extensive optimization and troubleshooting to obtain reliable data; this is further compounded by the mass of technical variations present throughout the literature. The aim of this article is to provide the necessary basics to get a quantitative real-time PCR assay up and running, and to address some of the problems that may arise and how these may be resolved. PMID- 17417023 TI - Protein extraction, fractionation, and purification from cyanobacteria. AB - This chapter deals with methods of protein extraction from cyanobacterial cells based on work in the circadian model organism Synechococcus elongatus PCC 7942. Some of these techniques have already been used successfully for analysis of circadian rhythms in cyanobacteria, whereas others are heretofore unpublished, but may yield exciting results in the near future. PMID- 17417024 TI - Protein extraction from Drosophila heads. AB - In Drosophila, the concentration and phosphorylation levels of several important circadian proteins (e.g., PERIOD, TIMELESS) oscillate on a 24-h basis. A simple and rapid method for extracting proteins from fly heads is presented here. The extracts can immediately be loaded onto an sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel to assay the effects of mutations, genetic manipulations, or environmental conditions on the oscillations of circadian proteins by Western blotting. They can also be used for immunoprecipitation experiments. PMID- 17417025 TI - Plant protein extraction. AB - A method is presented for the extraction of total protein from Arabidopsis thaliana tissue. The protocol was designed for the solubilization of a range of proteins and their efficient and quantitative recovery. It is especially compatible with the small quantities of available tissue often associated with this species and was originally intended for Western blot preparations. Samples extracted using this method can be quantitated directly using a commercially available kit. PMID- 17417026 TI - Protein extraction from mammalian tissues. AB - For Western blotting and coimmunoprecipitation (coIP), protein samples must be extracted from tissues. The protocol described in this chapter has been used to extract clock proteins from mammalian tissues as diverse as liver, kidney, and brain. The extraction protocol is mild enough to be used for coIP as well as Western blotting. Simply, clock proteins are extracted from tissues by freezing and thawing, and homogenizing with a handheld homogenizer. This procedure extracts most (>90%) of the clock proteins from mammalian tissues. PMID- 17417027 TI - Western blotting. AB - Western blotting is one of the most commonly used biochemical techniques to detect a specific protein from a mixture of proteins such as tissue extracts. Antibodies to the specific antigen are used to detect the protein. The mixture of proteins is resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a membrane. A specific antigen immobilized on the membrane is detected and visualized by a primary antibody, a secondary antibody-peroxidase conjugate, and a chemiluminescent reagent. PMID- 17417028 TI - Coimmunoprecipitation assay. AB - As with most other proteins, clock proteins physically interact with one another. Coimmunoprecipitation (coIP) is the most straightforward technique to study protein-protein interactions in vivo, if antibodies against the proteins of interest are available. To perform coIP, first an antibody against a target protein is coupled to Sepharose beads through protein A or G, then the complexes containing the target protein are immunoprecipitated with the antibody-coupled beads by centrifugation. Protein components in the complexes are visualized by Western blotting using antibodies specific to the different components. PMID- 17417029 TI - In vitro phosphorylation and kinase assays in Neurospora crassa. AB - Phosphorylation assay is a widespread technique usually necessary for the identification of a specific kinase substrate and/or for the measurement of kinase activity. As an example of the technique, here we describe an assay aimed to test the phosphorylation of the myelin basic protein (MBP) by protein kinase C (PKC), which is overexpressed and purified from Neurospora. The kinase is immunopurified from Neurospora using the expression vector pMYX2 and the FLAG epitope. The purified PKC and the MBP are then incubated in the presence of radioactive ATP, and the phosphorylated product is separated using the polyacrylamide gel electrophoresis technique. PMID- 17417030 TI - Basic protocols for Drosophila S2 cell line: maintenance and transfection. AB - Cells in culture have been increasingly employed in the dissection of intracellular processes. They are generally easier to handle than the organism of study and certainly less complex, which facilitates testing for specific functions and protein-protein interactions. This chapter will describe the extremely simple steps required to keep a healthy S2 cell culture going. PMID- 17417031 TI - Coimmunoprecipitation on Drosophila cells in culture. AB - Coimmunoprecipitation (coIP) provides evidence that two or more proteins can be found in the same complex. It can be performed in vitro (employing in vitro transcribed and translated proteins, or proteins expressed in Escherichia coli) or from transfected cells, which assess whether the interaction takes place in a more functional context. This chapter includes a general description and guidelines to carry out coIP in transfected Schneider's cells. PMID- 17417032 TI - Basic protocols for zebrafish cell lines: maintenance and transfection. AB - Cell lines derived from zebrafish embryos show great potential as cell culture tools to study the regulation and function of the vertebrate circadian clock. They exhibit directly light-entrainable rhythms of clock gene expression that can be established by simply exposing cultures to light-dark cycles. Mammalian cell lines require treatments with serum or activators of signaling pathways to initiate transient, rapidly dampening clock rhythms. Furthermore, zebrafish cells grow at room temperature, are viable for long periods at confluence, and do not require a CO2-enriched atmosphere, greatly simplifying culture conditions. Here we describe detailed methods for establishing zebrafish cell cultures as well as optimizing transient and stable transfections. These protocols have been successfully used to introduce luciferase reporter constructs into the cells and thereby monitor clock gene expression in vivo. The bioluminescence assay described here lends itself particularly well to high-throughput analysis. PMID- 17417033 TI - Manipulation of mammalian cell lines for circadian studies. AB - In mammals, the central circadian pacemaker resides in the hypothalamic suprachiasmatic nucleus (SCN), but circadian oscillators also exist in peripheral tissues. We have used wild-type and cryptochrome (mCry)-deficient mouse embryonic fibroblasts (MEFs) to demonstrate that the peripheral oscillator is mechanistically very similar to the oscillator in the SCN. Following serum shock activation, fibroblasts are able to sustain an SCN-like temporal expression profile of all known genes (i.e., antiphase oscillation of Bmal1 and Dbp genes), but are not able to produce oscillations in the absence of functional mCry genes. Remarkably, the analysis of mCry1-/- and mCry2-/- MEFs revealed the capacity to control period length in immortalized cell lines. Thus, the use of mammalian cells has become one of the most convenient methods for monitoring the molecular clock machinery and analyzing clock proteins at the functional/structural level. Here, we present the necessary protocols to (1) derive and culture a fibroblast cell line from wild-type and knockout mouse skin and (2) transfect cells at high efficiency to use in functional clock-protein studies. PMID- 17417034 TI - Reporter assays. AB - Transcriptional feedback loops are at the core of the molecular clockworks. As single clock genes were cloned it was compelling to develop an assay that allowed simple and direct functional testing of putative activators or repressors of transcription. This chapter includes a general description and guidelines to carry out transcriptional assays in transiently transfected Schneider's cells. PMID- 17417035 TI - Use of firefly luciferase activity assays to monitor circadian molecular rhythms in vivo and in vitro. AB - Circadian rhythms in metabolic, physiological, and behavioral processes are regulated by biological clocks. Many of these rhythmic processes can be measured over many days or weeks using automated recording devices, thus making it possible to precisely calculate period, phase, and amplitude values. With the advent of luciferase reporter genes and machines capable of quantifying luciferase-generated bioluminescence over long time frames, it is now possible to precisely monitor the rhythms in gene expression that underlie circadian clock function. These assays can be used to monitor gene expression in large numbers of individual plants and animals, and/or various cultured tissues and cells. After acquiring bioluminescence data, rhythm analysis programs are used to calculate the period, phase, amplitude, and overall levels of gene expression for individuals or groups, and to measure their statistical significance. Here we will describe how luciferase assays are performed and analyzed to measure gene expression rhythms in Drosophila. PMID- 17417036 TI - Suprachiasmatic nucleus cultures that maintain rhythmic properties in vitro. AB - Brain slices prepared from early postnatal rodents can be maintained in culture from many weeks to months. In culture, brain slices retain their original characteristic cytoarchitecture (organotypic) and continue to differentiate and mature in vitro resembling the characteristics of the original tissue in vivo. Therefore, this fascinating approach allows us to investigate fundamental issues of structure, function, and development of the central nervous system. This chapter introduces two techniques for culturing slices of mammalian brain tissue that are most commonly used at present. PMID- 17417037 TI - RNA in situ hybridizations on Drosophila whole mounts. AB - RNA in situ hybridization is a commonly used technique to achieve spatiotemporal detection of transcripts in tissues. This chapter gives an overview of novel techniques using fluorescent dyes, signal amplification methods, and confocal microscopy in regard to chronobiological applications on Drosophila adult brains. PMID- 17417038 TI - In situ hybridization of suprachiasmatic nucleus slices. AB - The progress in the understanding of the molecular machinery of mammalian circadian clocks, in combination with the well-established role of the hypothalamic suprachiasmatic nucleus (SCN) as a master circadian clock, has provided an invaluable system for the study of the molecular basis of circadian rhythmicity. Using in situ hybridization (ISH) techniques that label specific clock-gene mRNAs within the SCN, researchers can now elucidate the core molecular oscillatory mechanisms underlying specific circadian physiological and behavioral phenotypes. In this chapter, two methods for ISH within the SCN are described. The first method is based on the fluorescent labeling of mRNA and is suitable for confocal microscopy analysis and double labeling techniques. The second method is based on the radioactive labeling of mRNA and is more sensitive and more adequate for the relative quantification of mRNA species. PMID- 17417039 TI - Immunohistochemistry in Drosophila: sections and whole mounts. AB - This chapter describes immunohistochemistry on Drosophila heads (brains) with respect to antigens involved in the circadian system. Two different methods have been successfully performed in several labs: immunolabeling on whole-mount brains and on cryostat sections of entire heads. Both methods are addressed here. The primary antisera can be detected by enzyme-labeled or fluorescence-labeled secondary antibodies. The advantages of the different methods are discussed. PMID- 17417040 TI - Immunocytochemistry on suprachiasmatic nucleus slices. AB - Immunocytochemistry (ICC) is a sensitive and powerful method that is used to localize and identify cells containing a particular antigen. This chapter is dedicated to ICC of suprachiasmatic nucleus (SCN) slices. After a brief introduction to the technique, the materials and methods sections describe two different methods to obtain SCN slices--the first one for fixed tissue, the second one for fresh frozen tissue--followed by the description of two methods of antibody detection: the indirect method and the avidin-biotin complex one. In addition, some remedies to the most common problems encountered while performing ICC and some alternative protocols are discussed. PMID- 17417041 TI - Immunofluorescence analysis of circadian protein dynamics in cultured mammalian cells. AB - The timing of both entry and permanence of core-clock proteins in the nucleus is critical to maintain the correct pace of the clock mechanism. Several such proteins, namely CRYPTOCHROMEs (CRY), PERIODs (PER), and BMAL1, were recently shown to contain nuclear transport signals that facilitate their "shuttling" between the nucleus and the cytoplasm. This type of dynamic intracellular movement not only regulates protein localization, but also often affects functions by determining interactive partners and protein turnover. Because most clock genes have been identified by genetic screening in Drosophila and by gene knockdown in mammals, it is important to develop cellular techniques to study the structure-function and regulation of the corresponding proteins. Here we present working protocols for immunofluorescence studies of clock proteins in mammalian cultured cells. This technique allows the visualization in the cell of one or multiple proteins at the same time. PMID- 17417043 TI - Organizational and client determinants of cost in outpatient substance abuse treatment. AB - BACKGROUND: Understanding variation in the cost of outpatient substance abuse treatment is important for improving the delivery and financing of care. Studies that examine how the cost of treatment relates to treatment program and client characteristics can provide important data about variables that affect unit costs of treatment. Such analyses can inform those who are responsible for setting appropriate reimbursement rates and can give important cost data to program directors responsible for delivering cost-effective treatment. AIMS OF THE STUDY: The aim of this study is to describe the results from cost function analyses of outpatient substance abuse treatment programs sampled in the Alcohol and Drug Services Study (ADSS). The ADSS is a national study conducted in the late 1990s to collect organizational, client, and cost data of the specialty sector. METHODS: The authors examined how organizational and client characteristics affect the cost per episode and the cost per enrollment day of outpatient care. The analysis incorporates organizational variables such ownership, average length of stay, and visits per enrollment day, as well as client characteristics such as gender, age, and primary drug of choice. For further applicability for current treatment policy, the ADSS cost data were inflated from 1997 to 2005 dollars. Mixed model regressions using log-log and log-linear relationships were developed. RESULTS: Several organizational characteristics have statistically significant coefficients in the model estimating cost per episode, including log of point prevalence (-0.53, p<.01), log of average length of stay (0.73, p<.01), log of visits per enrollment day (0.45, p<.01), log of labor index (0.50, p<.01), proportion of counselor time spent in direct counseling (-0.52, p<.01), and location outside a metropolitan area (-0.19. p<.05). None of the client variables are statistically significant in this model. The analysis of cost per enrollment day indicates diseconomies of scope for programs that provide a broader array of ancillary services. DISCUSSION: Findings suggest there exist increasing returns to scale in outpatient substance abuse treatment. Mergers of substance abuse treatment programs may be economically beneficial. Other major determinants of cost include the average length of stay, wage rates, visits per enrollment day, and direct client contact time. Increased efficiency may enable programs to control costs in these areas. In addition, many of the patterns identified in the model represent the way in which outpatient substance abuse treatment facilities are reimbursed for services. As these patterns become more specified for client conditions, client factors may become statistically significant in determining costs. The potential problem of endogeneity is addressed. Limitations of the study include possible inaccuracies in non-personnel cost data, changes in the treatment system unaccounted for in the model, and limited market area information with regard to input prices. IMPLICATIONS FOR FURTHER RESEARCH: If further research indicates economies of scale, policymakers might consider supporting the merging of treatment programs. Also, further research into the optimal-mix of ancillary and treatment services would provide useful data for treatment programs seeking to balance resource constraints while providing important clinical and support activities. Lastly, research is needed to understand the relationship between treatment costs and service reimbursement. PMID- 17417044 TI - Racial differences in the use of adjunctive psychotropic medications for patients with schizophrenia. AB - BACKGROUND: Antidepressants, mood stabilizers and anxiolytics are frequently used in addition to antipsychotic medications in order to control ancillary symptom complexes among patients with schizophrenia. Although extensive data has demonstrated racial disparities in diagnostic and treatment patterns for schizophrenia, little is known about racial differences in the use of adjunctive medications. Since these adjunctive medications may significantly improve clinical outcomes, further investigation is warranted. AIMS OF THE STUDY: The purpose of this study was to determine the rates at which adjunctive antidepressants, mood stabilizers and anxiolytics were prescribed to black patients and white patients with schizophrenia. The authors hypothesized that black patients would be less likely to receive any adjunctive medication as compared to white patients. METHODS: This cross-sectional study utilized data from an academic medical center in Rochester, New York. Three multivariate logistic regression were performed to model the likelihood of receiving each of the three types of adjunctive medication. The primary independent variable was race, and the models also controlled for clinical and sociodemographic factors such as diagnosis, age, gender, living situation, and insurance status. RESULTS: Of the 456 patients included in the study, 37% received an antidepressant, 33% received a mood stabilizer and 23% received an anxiolytic. All patients received an antipsychotic medication. In each multivariate model, race was significantly associated with likelihood of receiving the adjunctive medication (p<.01 for all). None of the covariates was consistently associated with receipt of an adjunctive medication. DISCUSSION: Even after controlling for potentially confounding clinical and sociodemographic factors, race was strongly associated with receipt of adjunctive medications. This finding is consistent with previous work demonstrating racial disparities in the treatment and management of schizophrenia. The cross-sectional study design, however, limits the conclusions that can be drawn from this study, as length-time bias may have led to a misrepresentation of how many patients receive adjunctive medications. Caution should be observed in interpreting our results and replication is necessary. IMPLICATIONS FOR FURTHER RESEARCH: Differences in the use of adjunctive medications may be due to true differences in the frequency of ancillary symptom complexes. Previous work, however, suggests that differences are more likely to be due to underdiagnosis of affective or anxiety symptoms among black patients. Among patients with recognized ancillary symptoms complexes, black patients may also be less likely than white patients to receive treatment. This may be due to racial differences in accessibility of mental health care, physician perceptions of patients, and patient beliefs and preferences. In future work, use of standardized diagnostic tools may help to determine actual rates of ancillary symptom complexes and to identify racial variations. Additionally, further research is needed to clarify the underlying biases and behaviors that affect use of adjunctive medications among patients with schizophrenia in whom ancillary symptom complexes have been identified. PMID- 17417045 TI - Cost of schizophrenia in England. AB - BACKGROUND: Despite the wide-ranging financial and social burdens associated with schizophrenia, there have been few cost-of-illness studies of this illness in the UK. AIM OF THE STUDY: To provide up-to-date, prevalence based estimate of all costs associated with schizophrenia for England. METHODS: A bottom-up approach was adopted. Separate cost estimates were made for people living in private households, institutions, prisons and for those who are homeless. The costs included related to: health and social care, informal care, private expenditures, lost productivity, premature mortality, criminal justice services and other public expenditures such as those by the social security system. Data came from many sources, including the UK-SCAP (Schizophrenia Care and Assessment Program) survey, Psychiatric Morbidity Surveys, Department of Health and government publications. RESULTS: The estimated total societal cost of schizophrenia was 6.7 billion pounds in 2004/05. The direct cost of treatment and care that falls on the public purse was about 2 billion pounds; the burden of indirect costs to the society was huge, amounting to nearly 4.7 billion pounds. Cost of informal care and private expenditures borne by families was 615 million pounds. The cost of lost productivity due to unemployment, absence from work and premature mortality of patients was 3.4 billion pounds. The cost of lost productivity of carers was 32 million pounds. Estimated cost to the criminal justice system was about 1 million pounds. It is estimated that about 570 million pounds will be paid out in benefit payments and the cost of administration associated with this is about 14 million pounds. DISCUSSION: It is difficult to compare estimates from previous cost-of-illness studies due to differences in the methods, scope of analyses and the range of costs covered. Costs estimated in this study are detailed, cover a comprehensive list of relevant items and allow for different levels of disaggregation. The main limitation of the study is that data came from a variety of secondary sources and some official data publicly available was not the latest. IMPLICATIONS FOR HEALTH CARE PROVISION: Schizophrenia continues to be a high cost illness because of the range of health needs that people have. Despite the shifting balance of care away from hospital-based care, the health care costs of treating and supporting people with schizophrenia remain high. IMPLICATIONS FOR HEALTH POLICIES: Decision-makers need to recognise the breadth of economic impacts, well beyond the health system as conventionally defined. For example, as nearly 80% of schizophrenia patients remain unemployed, the cost of lost productivity is especially large. IMPLICATIONS FOR FURTHER RESEARCH: Better measurement of criminal justice services costs, private expenditures borne by families and valuation of lost quality of life could improve the estimates further. PMID- 17417046 TI - Migration, mental health and costs consequences in Romania. AB - BACKGROUND: Legal and illegal circulatory migration from Romania reached huge proportions after 2000, following the lifting of the visa requirements for EU Shengen countries. So far, the impact of migration on health has received scarce attention from Romanian authorities. AIMS OF THE STUDY: To describe the socio demographic and clinical profile of the migrants who have developed mental illness, estimate their services use in terms of hospitalization and to analyze the cost impact on the Romanian health system and on the migrants' co-payments, to discuss the possible relationships between migration and mental health. METHODS: A semi-structured interview, designed by the authors, has been administered to 50 migrants admitted to the Second Psychiatric Clinic Cluj Napoca, Romania, to investigate the following areas: immigration status, working conditions, income, housing, insurance and social bonds. The clinical symptomatology of these patients was assessed using the Brief Psychiatric Rating Scale (BPRS). The average cost of hospitalization per day per patient, the total costs of hospitalization and the migrants' co-payment through personal contribution to the insurance system were estimated. RESULTS: Most of the patients were young, single, with no previous experience abroad and with few social ties in the host country, with unqualified and insecure jobs. In this group, 45 out of 50 had schizophrenia spectrum disorders diagnoses. The hospitalization length of these patients was slightly shorter than the hospitalization of non-migrant patients with the same diagnosis. Individuals from rural areas had longer hospitalisation than those coming from urban areas. Those who left the country illegally and those who worked illegally had shorter hospitalisations. The average costs of hospitalization per day per patient were Euro 15.56; and the total costs were Euro 14,054.92. In order to cover the costs of hospitalization in the native country due to an illness with the onset abroad, a patient should work and contribute 4.65 years (on the basis of minimum salary) as a co-payment for the hospitalization in the native country. DISCUSSION: The patients considered in the present study fit the general profile of the emigrants. Discrimination, social isolation, insecurity may increase the risk for mental illness. A relevant role in the length of hospitalization may be due to the urban/rural dimension: patients from rural areas have less access to mental health care, therefore when they are finally hospitalised, they may remain longer because, in case of relapse, rapid readmission may not be possible. The shorter stay in hospital for those who left the country and worked illegally may be due to better resistance to stress and flexibility, to an illness that was not in an advanced phase, or to inability to afford higher personal co-payment for longer hospitalizaton. IMPLICATIONS FOR HEALTH POLICIES: More accurate and careful screening for mental illness should be applied at least for legal emigrants. Cultural adjustment programs should be organized prior to departure. Contact and counseling points in the host country would be important for prevention and treatment of mental illnesses. Programs focused on circulatory migrants and aimed at developing work opportunities in the native country would increase their self confidence and enable them to perceive their return as success and not as failure. PMID- 17417047 TI - EuroPRevent Congress, 19-21 April 2007, Madrid, Spain. Abstracts. PMID- 17417048 TI - Genome, proteome, and transcriptomes: the new systems approach to research. PMID- 17417049 TI - Mitochondrial dysfunction: impact on exercise performance and cellular aging. AB - Innovative noninvasive methods open a new window on the cell in vivo. This window reveals that the tempo of mitochondrial dysfunction with age varies among muscles and in proportion to Type II muscle fiber content. Exercise training can reverse age-related dysfunction, thereby providing an intervention to slow the pace of aging and disability in the elderly. PMID- 17417050 TI - Human muscle protein synthesis after physical activity and feeding. AB - The increase in protein synthesis after feeding is a systemic transient storage phenomenon, whereas physical exercise stimulates a local longer-term adaptive response. Providing nutrition after physical activity takes advantage of the anabolic signaling pathways that physical activity has initiated by providing amino acid building blocks and energy for protein synthesis. PMID- 17417051 TI - Primary cell cultures in the study of human muscle metabolism. AB - Skeletal muscle plays an important role in whole-body metabolism. Some research has used cell cultures raised from human biopsy tissue to study mechanisms that regulate skeletal muscle metabolism. The purpose of the current paper is to provide evidence indicating the efficacy of primary human skeletal muscle cell cultures as a tool to study substrate regulation and control in human tissue. PMID- 17417052 TI - Influence of racket properties on injuries and performance in tennis. AB - Racket properties may affect tennis elbow development. Shock and vibration to the arm is influenced by the location of ball impacts on the racket head, racket stiffness, and grip force. Beginners experience increased arm loads, and they hit the ball lower on the racket head. Tennis rackets behave differently during actual play compared with the performance predicted by physics. PMID- 17417053 TI - Development of independent walking in toddlers. AB - Surprisingly, despite millions of years of bipedal walking evolution, the gravity related pendulum mechanism of walking does not seem to be implemented at the onset of independent walking, requiring each toddler to develop it. We discuss the precursor of the mature locomotor pattern in infants as an optimal starting point strategy for gait maturation. PMID- 17417054 TI - Muscle power: the interaction of cycle frequency and shortening velocity. AB - Force-velocity, force-time, and force-length relations of muscle are typically evaluated with reductionist techniques in which force is measured while the variable of interest is manipulated. However, force produced during voluntary movement arises from complex interaction of these contractile characteristics. Nonetheless, neuromuscular performance for cyclical, work-producing movements can be predicted from the interaction of average muscle shortening velocity and movement frequency. PMID- 17417055 TI - The PDE4B gene confers sex-specific protection against schizophrenia. AB - BACKGROUND: Phosphodiesterase 4B (PDE4B) is a candidate gene for schizophrenia and affective disorders through its disruption by a chromosomal translocation in an individual with schizophrenia, its inhibition by the antidepressant rolipram, and its physical interaction with another key candidate, Disrupted in Schizophrenia (DISC1). OBJECTIVE: To determine the contribution made by PDE4B to the population risk of schizophrenia and bipolar disorder by carrying out a case control association study. METHODS: Twenty-six tagging single nucleotide polymorphisms were selected across the PDE4B gene and genotyped in DNA samples from 386 schizophrenia cases, 368 bipolar disorder cases and 455 controls. MAIN RESULTS: Single single nucleotide polymorphisms and a resulting haplotype conferred a protective effect against schizophrenia in the female population. The haplotype result remained significant after correction for multiple testing (P=0.012). CONCLUSION: The observation that a PDE4B haplotype alters the genetic risk of schizophrenia in the Scottish population complements the known participation of this gene in biological processes associated with mental illness. Further studies are needed to replicate this finding and identify underlying sequence variants. PMID- 17417056 TI - Temperamental fearfulness in childhood and the serotonin transporter promoter region polymorphism: a multimethod association study. AB - OBJECTIVES: Early-emerging, temperamental differences in fear-related traits may be a heritable vulnerability factor for anxiety disorders. Previous research indicates that the serotonin transporter promoter region polymorphism is a candidate gene for such traits. METHODS: Associations between 5-HTTLPR genotype and indices of fearful child temperament, derived from maternal report and standardized laboratory observations, were examined in a community sample of 95 preschool-aged children. RESULTS: Children with one or more long alleles of the 5 HTTLPR gene were rated as significantly more nervous during standardized laboratory tasks than children who were homozygous for the short alleles. Children homozygous for the short alleles were also rated as significantly shyer, by maternal report, than those with at least one copy of the long allele of the 5 HTTLPR gene. CONCLUSIONS: This study extends the literature linking the short alleles of the serotonin transporter promoter region polymorphism to fear and anxiety-related traits in early childhood and adulthood, and is one of very few studies to examine the molecular genetics of preschoolers' temperament using multiple measures of traits in a normative sample. PMID- 17417057 TI - The Gilles de la Tourette syndrome: a principal component factor analytic study of a large pedigree. AB - BACKGROUND: The genetics and phenotypes of Gilles de la Tourette syndrome are complicated. Once indicated to be inherited as a single major autosomal dominant condition, several areas of interest on many chromosomes and one gene have been identified for Gilles de la Tourette syndrome, but no results have been replicated. Factor analytic studies suggest that there are more than one Gilles de la Tourette syndrome phenotype and it is not a unitary condition. OBJECTIVE: To characterize Gilles de la Tourette syndrome phenotypes in a group of individuals who underwent a complete genome scan. METHODS: We studied 85 members of a multiply affected multigenerational kindred, of whom 69 displayed Gilles de la Tourette syndrome-related symptoms (tics, obsessive-compulsive behaviours, obsessive-compulsive symptoms, attention deficit hyperactivity symptoms), using first a hierarchical cluster analysis followed by a principal component factor analysis. RESULTS: Three significant factors resulted from our analysis, accounting for approximately 42% of the symptomatic variance: Factor 1 (predominantly 'pure tics'), Factor 2 (predominantly 'attention deficit hyperactivity disorder and aggressive behaviours') and Factor 3 (predominantly 'depression-anxiety-obsessional symptoms and self-injurious behaviours'). Different kinds of tics occurred in all three factors. Only frowning/raising eyebrows and sniffing/smelling loaded significantly on both Factors 1 and 3. CONCLUSION: Our results give further evidence that the genetics of Gilles de la Tourette syndrome is complex and suggest that Gilles de la Tourette syndrome is not a unitary condition, thus confirming the results of earlier studies which have described several Gilles de la Tourette syndrome phenotypes. Although a genome scan on the pedigree reported three areas of interest and the present study found three factors, further studies would have to be undertaken to elucidate whether the three factors 'mapped' with the genetic data. Possible reasons for our findings and suggestions for future research are discussed. PMID- 17417058 TI - Monoamine oxidase a gene is associated with borderline personality disorder. AB - OBJECTIVE: Monoamine oxidase A is a mitochondrial enzyme involved in the degradation of certain neurotransmitter amines: serotonin and norepinephrine. As for its role in aggression, impulsivity, suicide and mood liability, monoamine oxidase A can be considered a functional candidate in borderline personality disorder. METHODS: To test for this hypothesis we genotyped two polymorphic markers in monoamine oxidase A gene, a promoter VNTR and an rs6323 (T941G) in exon 8, in 111 Caucasian borderline personality disorder patients and 289 Caucasian healthy controls. Association analyses using individual marker and haplotype data were performed by a program of COCAPHASE in UNPHASED (MRC Human Genome Mapping Project Resource Centre, Cambridge, UK). RESULTS: We found that the borderline personality disorder patients had a high frequency of the high activity VNTR alleles (chi=4.696, P=0.03) and a low frequency of the low activity haplotype (chi=5.089, P=0.02). CONCLUSION: These results show that the monoamine oxidase A gene may play an important role in the etiological development of the borderline personality disorder. PMID- 17417059 TI - A dopamine D2 receptor gene-related polymorphism is associated with schizophrenia in a Spanish population isolate. AB - Numerous lines of evidence have highlighted the involvement of the dopamine system in the pathophysiology of schizophrenia. Association studies of dopaminergic genes such as the dopamine D2 receptor gene (DRD2), however, have produced contradictory results. To test the hypothesis that DRD2 polymorphisms are associated with schizophrenia, we investigated two DRD2-related polymorphisms (TaqI A1/A2 or rs1800497 and -141-C Ins/Del or rs1799732) in a Spanish population isolate from northern Spain consisting of 165 controls and 119 patients with schizophrenia. The TaqI A1 allele was less frequent in schizophrenic patients than in controls (P=0.002). A similar association was found for the TaqI A2/A2 genotype (P=0.0003). No association was found for the DRD2 -141-C Ins/Del polymorphism. The strong association between a potentially functional polymorphism, downstream of the DRD2 gene and schizophrenia, suggests that the direct or indirect functional effects of this polymorphism, acting on either the ANKK1 or DRD2 genes, may play a role in the pathophysiology of schizophrenia. PMID- 17417060 TI - Meta-analysis reveals no association of the Val66Met polymorphism of brain derived neurotrophic factor with either schizophrenia or bipolar disorder. AB - BACKGROUND: A long-term controversy exists on whether or not major psychotic disorders can be discretely divided into two groups, for example, schizophrenia and bipolar disorder. Many genes and polymorphisms have been studied for a role in both disorders, including the Val66Met (also known as rs 6265 or G196A) variant of brain-derived neurotrophic factor (BDNF). Many case-control association studies have been performed to see if BDNF could serve as a useful clinical diagnostic biomarker for schizophrenia or bipolar disorder, but results have been equivocal. OBJECTIVE: To determine, by meta-analysis, if the Val66Met polymorphism of BDNF influences risk for either schizophrenia, bipolar disorder, or both. METHODS: We searched Pubmed, Medline, and PsycInfo using keywords including Val66Met, Rs6265, G196A, BDNF, schizophrenia, and bipolar disorder. A total of 13 studies for schizophrenia and 11 studies for bipolar disorder were combined by random-effects meta-analysis. MAIN RESULTS: The pooled results from the schizophrenia sample (2955 patients; 4035 controls) and the bipolar disorder sample (3143 patients; 6347 controls) indicated lack of significance with either of the two psychoses, with pooled odds ratios of 1.00 (P=0.944) and 0.95 (P=0.161), respectively. CONCLUSION: Although there are some limitations on the study, our results indicate there is a lack of association between the Val66Met polymorphism and either of the two psychoses. A larger sample size, and evaluation of more single-nucleotide polymorphisms are needed to obtain more robust and conclusive findings regarding the relationship between the BDNF gene and psychosis. PMID- 17417061 TI - Heritability of illicit drug use and transition to dependence in Southwest California Indians. AB - OBJECTIVE: Native Americans have high rates of drug use and dependence yet little is known concerning its etiology or clinical course. These analyses were conducted to describe the heritability of the use of a variety of illicit drugs, as well as the conditional probability of transitioning from use to dependence for each drug class in a community sample of Native American men and women. METHODS: The sample included 460 participants (190 men and 270 women), recruited through community effort, from eight contiguous Indian reservations in Southern California. Participants were assessed using the Semi-Structured Assessment for the Genetics of Alcoholism. The Semi-Structured Assessment for the Genetics of Alcoholism interview retrospectively asks about the initial use and drug dependence of the following illicit drug classes: marijuana, cocaine, stimulants, sedatives, opiates, hallucinogens, and solvents. Heritability of initial use was determined using SOLAR (http://www.sfbr.org/solar/). RESULTS: Ninety-one percent of this select Indian population had tried at least one of the illicit drug classes. The most commonly tried substance was marijuana (88%), followed by stimulants (60%), cocaine (44%), hallucinogens (34%), and solvents (20%). The heritability of initiation of drug use ranged from 0.14 for cocaine to 0.59 for marijuana. The conditional probability of transition from initiation to drug dependence ranged from 0.66 for stimulants to 0.06 for hallucinogens. CONCLUSIONS: These findings suggest that heritability of the initiation of substance use, in Southwest California Indians, may be similar to other population samples. In this population, however, high rates of dependence on marijuana, opiates, and stimulants are seen once initiation of the use of the substance has occurred. PMID- 17417062 TI - MTHFD 1958G>A and MTR 2756A>G polymorphisms are associated with bipolar disorder and schizophrenia. AB - BACKGROUND: The etiology of bipolar disorder (BD) and schizophrenia is very complex. Polymorphic variants of genes encoding enzymes of the monoaminergic may be involved in development of BD and schizophrenia. Therefore, we examined the prevalence of 1958G>A polymorphism of MTHFD1 gene, encoding trifunctional folate enzyme 5,10-methylenetetrahydrofolate dehydrogenase, 5,10 methenyltetrahydrofolate cyclohydrolase and 10-formyltetrahydrofolate synthetase (MTHFD1), and 2756A>G variant of methionine synthase (MTR) gene in patients with BD (n=200), schizophrenia (n=200) and in controls (n=300). OBJECTIVE: We investigated the genotypic and allelic frequencies of MTHFD1 1958G>A (R653Q) and MTR 2756A>G (D919G) gene polymorphisms in a group of bipolar (n=200) and schizophrenic patients (n=200), as well as in controls (n=300). METHODS: The distributon of genotypes in all groups was tested for deviation from Hardy Weinberg equilibrium (HWE). The Pearson's chi-square (chi) test and Fisher's exact test were applied to assess differences in the genotypic and allelic (respectively) distribution between groups of patients and controls. MAIN RESULTS: We found that MTHFD1 1958AA or 1958AG genotypes constitute risk factors for development of bipolar disorder type I (BDI) or schizophrenia with odds ratios (OR)=1.743 (95% CI=1.211-2.508; P=0.0027; P (corr)=0.0054) and 2.667 (95% CI=1.845-3.854; P=0.0001; P (corr)=0.0002), respectively. In the same groups, the MTR 2756GG or 2756AG genotypes also constitute significant risk factors in occurrence of BDI and schizophrenia with OR=1.621 (95% CI=1.130-2.326; P=0.0086; P (corr)=0.0172) and 1.556 (95% CI=1.085-2.232; P=0.0160; P (corr)=0.032), respectively. Gender classification of patients indicated significant association only of MTHFD1 1958A allele with BDI and schizophrenia in the male patients OR=1.838 (95% CI=1.114-3.031; P=0.0166; P (corr)=0.0332) and OR=3.964 (95% CI=2.358-6.663; P=0.0001 P (corr)=0.0002), respectively. CONCLUSION: Since MTHFD and MTR genes are located in 14q24 and 1q43 loci, our findings support the significance of chromosomes 14q and 1q in etiopathogenesis of bipolar disorder and schizophrenia. PMID- 17417063 TI - Catechol-o-methyltransferase polymorphism and susceptibility to major depressive disorder modulates psychological stress response. AB - OBJECTIVES: The stress response is related to both physiological and psychological factors and is strongly marked by a neuroendocrine component. Genetic factors are believed to underlie individual differences in the degree of stress resilience and thereby contribute in determining susceptibility to stress related pathologies like major depressive disorder (MDD). Little, however, is known about the genetic influence on the endocrine and behavioural stress response in relation to MDD. METHODS: Here, we sought to examine the effects of the catechol-o-methyltransferase polymorphism on psychological stress in three groups of individuals with different degrees of susceptibility to MDD (i.e. healthy controls, healthy high risk probands to MDD and those suffering from MDD). This genotype is involved in the metabolism of catecholamines (dopamine, norepinephrine and epinephrine). RESULTS: Allelic variations of this polymorphism were found to influence the degree of subjective stress experience and plasma epinephrine stress response. Interactions between catechol-o-methyltransferase polymorphism and diagnostic group in measures of plasma epinephrine, cortisol and subjective responses to psychological stress were also found, with the influence of the different alleles on these measures differing between healthy controls relative to MDD patients and high risk probands. CONCLUSION: These observations support a possible role for catechol-o-methyltransferase polymorphism in the endocrine and subjective response to psychological stress and thus may qualify as a possible candidate gene involved in the pathogenesis of MDD. PMID- 17417064 TI - Genetic pathway of major depressive disorder in shortening telomeric length. AB - CONTEXT: Shortened telomeric length has been associated with aging and monoamine oxidase A gene activity. The pathways of genetic activity and mental disorder in shortening telomeric length, however, remain unclear. OBJECTIVE: The purpose of this study was to explore the possible pathways of the monoamine oxidase A promoter, ApoE polymorphisms and telomeric length in major depressive disorder. METHODS: This study enrolled 253 unrelated patients with major depression in southern Taiwan, and was carried out between March 2001 and September 2004. In addition, 411 controls were randomly selected from the general population in southern Taiwan. RESULTS: Hierarchical regression showed that the influence of the monoamine oxidase A promoter and ApoE2 polymorphisms was not statistically significant to telomeric length, when additionally adjusting for major depressive disorder. A final robust structural equation model showed that aging and major depressive disorder both have a statistically significant shortening effect on telomeric length. Moreover, sex, the Apoepsilon2 allele, and the monoamine oxidase A promoter polymorphism have indirect effects on telomeric length. CONCLUSION: Major depressive disorder is a mediating factor between the monoamine oxidase A promoter polymorphism and telomeric length. PMID- 17417065 TI - Polymorphisms of the Fyn kinase gene and a performance on the Wisconsin Card Sorting Test in schizophrenia. AB - The glutamatergic system has been implicated in the pathogenesis and prefrontal cortex dysfunctions in schizophrenia. The Src-family tyrosine kinase Fyn plays a key role in the interaction between brain-derived neurotrophic factor and glutamatergic receptor N-methyl-D-aspartate, in prefrontal cortex. We estimated an association between three polymorphisms of Fyn gene and performance on the Wisconsin Card Sorting Test, measuring prefrontal cortex functions, in 188 schizophrenic patients. Patients with T/T genotype of IVS10+T/C polymorphism and T/T genotype of Ex12+894T/G polymorphism made significantly less perseverative errors in the Wisconsin Card Sorting Test compared with patients with remaining genotypes, and obtained numerically better results in other Wisconsin Card Sorting Test domains. No significant differences in Wisconsin Card Sorting Test performance were found as to -93 A/G polymorphism. The main finding of the study is showing a relationship between polymorphisms of the Fyn gene, related to the function of glutamatergic system, and a performance on neuropsychological test of prefrontal cortex activity in schizophrenic patients. PMID- 17417066 TI - A case-control study of genetic association of the PLA2G4A gene with schizophrenia in a Chinese population. PMID- 17417067 TI - Monitoring of inosine monophosphate dehydrogenase activity as a biomarker for mycophenolic acid effect: potential clinical implications. AB - Mycophenolic acid (MPA) is a reversible inhibitor of inosine monophosphate dehydrogenase (IMPDH) and, in combination with other immunosuppressive drugs, effectively inhibits rejection in solid organ transplant recipients. MPA has a relatively narrow therapeutic window and exhibits wide inter- and intrapatient pharmacokinetic (PK) variability. This has stimulated the use of therapeutic drug monitoring as a strategy to tailor the MPA exposure to each patient's individual needs. Despite increasing therapeutic drug monitoring use, PK-assisted dosing is not universally adopted in part because of MPA's complex PK behavior. Targeting inosine monophosphate IMPDH activity as a surrogate pharmacodynamic (PD) marker of MPA-induced immunosuppression may allow for increased precision when used in an integrated PK-PD fashion, providing a more accurate assessment of efficacy and aid in limiting toxicity. IMPDH activity displays wide interpatient variability but relatively small intrapatient variability even after long-term administration of MPA. The advent of calcineurin and corticosteroid-sparing regimens necessitates more patient-specific PK-PD parameters, which can be used throughout the posttransplant period to optimize MPA exposure and immediate and long-term graft and patient outcomes. Quantification of IMPDH posttransplant may serve as a stable, surrogate PD marker of MPA-induced immunosuppression when combined with current PK and monitoring strategies. PMID- 17417068 TI - Analytical methods of biological monitoring for exposure to pesticides: recent update. AB - Extensive use of synthetic pesticides for agricultural and nonagricultural purposes began in the past 50 years. As a result of their wide and extensive application, exposure to hazardous pesticides is a concern to the general population and occupationally exposed persons. Robust methods are therefore needed for measuring markers of pesticide exposure. This article presents a review of the most recently published analytical methodologies and instrumentations developed for and applied to biological monitoring of exposure to pesticides of various classes. Most of the methods reviewed here are based on chromatography combined with mass spectrometry detection. This work clearly demonstrates that although gas chromatography still appears to be the most widely employed technique for pesticide analysis in various biological samples, recently a trend has been observed toward the use of liquid chromatography coupled with tandem mass spectrometry. PMID- 17417069 TI - Indinavir trough concentration as a determinant of early nephrolithiasis in HIV-1 infected adults. AB - Indinavir plasma levels are associated with antiretroviral efficacy; however, little data are available regarding toxicity. We assessed the relationship between indinavir pharmacokinetic (PK) characteristics and severe nephrolithiasis as well as other severe or serious adverse reactions. Patients included in the ANRS CO8 APROCO-COPILOTE cohort and receiving 800 mg indinavir three times daily as a first-line protease inhibitor were eligible for this study. To be included in the analysis, their plasma sample at month 1 (M1) had to be available (n = 282) to estimate using population PK modeling, indinavir PK characteristics, ie, maximum (Cmax) and trough plasma (Cres) concentrations, area under the curve (AUC), and observed/predicted concentration ratio (CR). A Cox model was used to estimate the independent effect of Cmax, Cres, AUC, and CR on the hazard of severe nephrolithiasis and serious adverse reactions. At M1, median Cmax was 6205 ng/mL, Cres 631 ng/mL, AUC 24,242 ng . h/mL, and CR 0.6. After a median follow up of 12 months, 11% of patients (30 of 282) had experienced at least one serious adverse reaction among which 12 were nephrolithiasis. In the multivariate analyses, early high indinavir Cres (ie, >/=1000 ng/mL at M1) was associated with a higher rate of severe nephrolithiasis (hazard ratio = 6.7; 95% confidence interval = 1.8-25.2; P < 0.01) and was also associated with a higher rate of all serious adverse reactions but only when nephrolithiasis were included among those cases. Prospective and early indinavir Cres determination should be recommended in the patient's care management and dosage adjustments. PMID- 17417070 TI - Efficacy of low-dose boosted saquinavir once daily plus nucleoside reverse transcriptase inhibitors in pregnant HIV-1-infected women with a therapeutic drug monitoring strategy. AB - The efficacy of low-dose, ritonavir-boosted saquinavir (SQV/rtv) once daily plus 2 nucleoside retrotranscriptase inhibitors (NRTIs) in pregnant human immunodeficiency virus (HIV)-1-infected women was prospectively evaluated, ensuring a SQV minimum concentration (Cmin) >/=100 ng/mL with a therapeutic drug monitoring strategy. The primary clinical endpoint was the percentage of women with an HIV-RNA viral load (VL) of <50 copies/mL at the time of delivery. Forty nine pregnancy episodes were included, with a median CD4 count and VL of 441/muL and 3710 copies/mL, respectively. Two patients were lost to follow-up and 1 patient discontinued treatment because of abdominal discomfort. SQV levels were in excess of the target Cmin in 43 of 46 episodes (93.4%) in which the end of pregnancy was reached on 1200/100 mg daily. The dosage was increased to 1600/100 mg in the remaining 3 episodes to achieve the target levels. By an intention-to treat analysis, VL was undetectable at delivery in 43 episodes (87.7%; 95% confidence interval, 78.5-96.9) after a median of 18 weeks of treatment (range, 3 39). In the 3 episodes remaining, VLs of 110,400 copies/mL and no available data were observed after only 3 weeks of treatment. Mild adverse events attributable to SQV/rtv occurred in 6 of 49 pregnancies (12.2%). No cases of HIV vertical transmission were observed. The pharmacokinetics, efficacy, and tolerability of this regimen suggest that once-daily low-dose boosted SQV may be considered an appropriate option in PI-naive or limited-PI-experienced HIV-infected pregnant women. Nevertheless, therapeutic drug monitoring is advisable to maintain appropriate levels throughout pregnancy. PMID- 17417071 TI - Influence of underlying disease on busulfan disposition in pediatric bone marrow transplant recipients: a nonparametric population pharmacokinetic study. AB - Busulfan is an alkylating agent used in a conditioning regimen prior to bone marrow transplantation. Busulfan has a narrow therapeutic index, giving rise to major liver toxicity (veno-occlusive disease), and a wide interpatient and intrapatient pharmacokinetic variability. This report presents the results of a population pharmacokinetic analysis leading to models based on underlying diseases requiring bone marrow transplantation. One hundred children received oral busulfan-based conditioning regimens between March 1998 and February 2006. Busulfan pharmacokinetic parameter estimates (Ka, first order absorption rate constant; Vs, volume of distribution related to the body weight; and Cl/F, apparent clearance) were estimated by using the nonparametric adaptative grid (NPAG) algorithm in patients divided into four groups according to initial diagnosis: metabolic diseases, hemoglobinopathies, hematological malignancies, and immune deficiencies. Ka and Vs did no differ significantly in the four subgroups. Cl/F and areas under the plasma concentration curve were significantly different in the four groups. Cl/F was significantly higher in the hemoglobinopathies group (P = 0.002), with a mean value of 7.78 L . h, whereas the immune deficiencies group was characterized by the lowest Cl/F (3.59 L . h). Interindividual variability was shown by high interindividual parameter percent coefficients of variation (CV%) but, nevertheless, with less diversity in the population parameter distributions for Vs in the three subgroups-metabolic diseases, hemoglobinopathies, and malignant diseases-and in Cl/F for patients with hemoglobinopathies. The fit was good for busulfan concentration predictions based on Bayesian individual posterior values, with little bias and good precision. In comparison with the overall population, the only model of subgroup presenting a greater precision was patients with hemoglobinopathies (P = 0.002). Use of these more specific models of a given disease may well result in more accurate individualization of busulfan dose regimens, especially in very sparse blood sampling situations. PMID- 17417072 TI - Dose-dependent effects of the 3435 C>T genotype of ABCB1 gene on the steady-state plasma concentration of fluvoxamine in psychiatric patients. AB - This study investigated effects of the 3435 C>T genotype of the adenosine triphosphate-binding cassette subfamily B member 1 (ABCB1, MDR1) gene on the steady-state plasma concentration of fluvoxamine (FLV). METHODS: Sixty-two psychiatric patients were treated with different doses (50, 100, 150, and 200 mg/d) of FLV. Blood samples were collected after at least 2 weeks of treatment with the same daily dose to obtain steady-state concentrations of FLV, and 3435 C>T genotype was determined by polymerase chain reaction. RESULTS: FLV concentration-to-dose ratio was significantly different among 3435 C>T genotype groups at the 200 mg/d dose (P = 0.019). A post-hoc analysis revealed that FLV concentration-to-dose ratio was significantly higher in the TT genotype group as compared with the CC genotype group at the 200 mg/d dose (median value of concentration-to-dose ratio (ng/mL)/(mg/d), 0.861 vs 0.434, P = 0.026). FLV concentration-to-dose ratio was significantly higher in the CT + TT genotype group than the CC genotype group at the 200 mg/d dose (median value of concentration-to-dose ratio (ng/mL)/(mg/d), 0.618 vs 0.434, P = 0.031). At 50, 100, and 150 mg/d dose, FLV concentration-to-dose ratios were not significantly different among 3435 C>T genotype groups. At 50, 100, and 150 mg/d dose, no significant differences were found in FLV concentration-to-dose ratios between the CT + TT genotype group and CC genotype group. CONCLUSIONS: This study suggests that pharmacokinetics of FLV depend on ABCB1 gene polymorphism only at the 200 mg/d dose. PMID- 17417073 TI - Thymidylate synthase (TYMS) and dihydropyrimidine dehydrogenase (DPYD) polymorphisms in the Korean population for prediction of 5-fluorouracil associated toxicity. AB - The important cellular proteins for 5-fluorouracil (5-FU) metabolism are the major target enzymes, thymidylate synthase, and the rate-limiting enzyme in the degradation pathway, dihydropyrimidine dehydrogenase. Adverse drug reactions to 5 FU-based chemotherapy have been reported to be in part the result of polymorphisms in the thymidylate synthase (TYMS) and dihydropyrimidine dehydrogenase (DPYD) genes. Therefore, we examined the type and frequency of polymorphisms in the TYMS and DPYD genes in 100 healthy Korean individuals and compared these findings with 21 patients with colorectal cancer who had a grade 3 or greater toxic response to 5-FU treatment. Genotyping analysis of the promotor enhancer region (TSER) and the 3'-untranslated region (3'-UTR) of the TYMS gene as well as haplotype analysis were conducted in all 121 study participants. For the TSER and the 3'-UTR of the TYMS gene, similar genotypes and allele frequencies were observed in control subjects and patients. For the haplotype analysis of the single nucleotide polymorphism G > C at the 12th nucleotide of the second repeat of the 3R allele of the TSER, different haplotype frequencies were noted in comparisons between the two groups; in addition, we found that the 3RC-del 6bp was significantly associated with severe toxicity with 5-FU treatment. Extensive polymorphisms in the DPYD gene were observed; in addition, four polymorphisms were related to the known DPYD allelic variants or to allelic variants that alter protein structure, among which the most common polymorphism was 1627A > G, observed in 20.5% of all alleles. The 496A > G allele and a novel 1774C > T allele were identified in two patients. The DPYD*2A allele, causing exon 14 skipping, was not identified in the study group. The findings, from Korean patients with colon cancer, suggest that polymorphisms of the DPYD gene are not associated with an increased risk for toxic response to 5-FU. These findings suggest that there may be an important relationship between the TYMS haplotypes examined and 5-FU toxicity. The novel variant in the DPYD gene, identified in this study, should be further investigated to confirm its functional significance. A large sample is required before DPYD or TYMS genotyping could be used as markers for individualized treatment of patients with colorectal cancer. PMID- 17417074 TI - Determination of fenofibric acid concentrations by HPLC after anion exchange solid-phase extraction from human serum. AB - Triglycerides are increasingly being recognized as a risk factor for cardiovascular disease. Research efforts to identify sources of variability in triglyceride-lowering response to the lipid-lowering drug fenofibrate require quantification of the active acidic form of this PPAR-alpha agonist. Anion exchange solid-phase extraction, in combination with reverse-phase high performance liquid chromatography (HPLC), rapidly and accurately determines steady-state fenofibric acid serum concentrations. Chromatographic separation under isocratic conditions, with use of ultraviolet detection at 285 nm, provides clean baseline and sharp peaks for clofibric acid, 1-napthyl acetic acid (internal standards), and fenofibric acid. Commonly prescribed and over-the counter nonsteroidal anti-inflammatory drugs (NSAIDs) were screened for assay interference, and the assay was employed to quantify fenofibric acid in more than 800 human subject specimens. Fenofibric acid analysis was found to be linear over the range of 0.5 to 40 mg/L and was validated with either internal standard. Accuracies ranged from 98.65% to 102.4%, whereas the within- and between-day precisions ranged from 1.0% to 2.2% and 2.0% to 6.2%, respectively. NSAIDs had minimal interference with the assay, which succeeded in quantifying fenofibric acid in more than 843 of 846 serum samples from human subjects, many taking a variety of coadministered medications. Anion-exchange solid-phase extraction in combination with reverse-phase HPLC accurately determines steady-state fenofibric acid serum concentrations in humans without interference from NSAIDs or commonly administered medications. This method is suitable for quantification of fenofibric acid for clinical pharmacokinetic studies in patients with dyslipidemia. PMID- 17417075 TI - Effects of prior administration of amodiaquine on the disposition of halofantrine in healthy volunteers. AB - The prevalence of multidrug-resistant malaria parasites brings about the switch from an antimalarial drug with poor therapeutic outcome to an effective alternative, resulting in overlap in the plasma drug levels. In this study, the influence of prior administration of amodiaquine on the pharmacokinetics and electrocardiographic effect of halofantrine (HF) was investigated in healthy volunteers. Ten healthy male subjects were each given single oral doses of 500 mg HF alone or with 600 mg of amodiaquine hydrochloride (AQ) administered 24 hours before the HF dose in a crossover fashion. Blood samples, collected at predetermined time intervals, were analyzed for HF and its major metabolite, desbutylhalofantrine (HFM) using a validated high-performance liquid chromatography method. Electrocardiogram for each volunteer was taken at predetermined time points. Results showed that prior administration of amodiaquine resulted in no significant changes (P > 0.05) in any of the pharmacokinetic parameters of HF. For example, the parameter values for HF alone and with AQ were: Cmax 144 +/- 53 versus 164 +/- 58 microg/L; T1/2beta 142 +/- 23 versus 139 +/- 28 hours; Cl/F 37.3 +/- 13.9 versus 32.3 +/- 11.4 L/h; and metabolic ratio 1.2 +/- 0.5 vs 1.1 +/- 0.6 Similarly, the disposition of HFM was not significantly altered (P > 0.05) after an earlier exposure to amodiaquine. In addition, the presence of AQ was linked with a further lengthening of the QT interval compared with the effect of HF alone. This study suggests that prior administration of AQ does not result in a significant alteration of the pharmacokinetics of HF but may be associated with an increased risk of QT prolongation. It may be necessary to exercise caution in the use of HF for malaria treatment in persons who have recently received AQ. PMID- 17417076 TI - Limited sampling strategy for the estimation of mycophenolic acid area under the plasma concentration-time curve in adult patients undergoing liver transplant. AB - Mycophenolate mofetil (MMF), the oral prodrug of mycophenolic acid (MPA), is increasingly used in liver transplantation and plays a central role in the immunosuppressive regimen in liver transplantation. To study pharmacokinetic pharmacodynamic relationships and therapeutic drug monitoring of MPA in the clinical setting, limited sampling strategies have been investigated for the estimation of MPA areas under the curves (AUCs). Thirty-eight adult patients undergoing liver transplant (31 males, seven females) receiving 1.0 g MMF twice daily and concomitant tacrolimus provided a total of 72 pharmacokinetic profiles. Multiple stepwise regression analysis was used to determine the algorithms for limited sampling strategies. Twenty-eight one-, two-, three-, and four-sampling estimation models were fitted (r = 0.288-0.964) to all the profiles using linear regression and were used to estimate MPA AUC0-12h comparing those estimates with the corresponding AUC0-12h values calculated with the linear trapezoidal rule, including all 10 timed MPA concentrations. The four-point estimates at C1h, C2h, C6h, and C8h resulted in the best correlation between estimated AUC and true AUC when using the formula AUC = 6.03 + 0.89C1h + 1.94C2h + 2.24C6h + 4.64 C8h (r = 0.911). Bland and Altman analysis revealed good agreement between estimated AUC and AUC from the full profile. This limited sampling strategy provides an effective approach for estimation of full MPA AUC0-12h in patients undergoing liver transplant receiving concomitant tacrolimus therapy. PMID- 17417077 TI - Determination of free and total cortisol in plasma and urine by liquid chromatography-tandem mass spectrometry. AB - Cortisol is an important adrenal steroid hormone involved in the regulation of metabolic homeostasis. A new liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) multiple reactant monitoring (MRM) procedure for the measurement of cortisol concentration in plasma ultrafiltrate, whole plasma, and urine was developed and validated. Plasma, plasma ultrafiltrate, or urine was extracted by ethyl acetate. The extract was subjected to liquid chromatography with an Inertsil ODS-3 column with an aqueous NH4Cl (1 mM, pH 9.0):methanol mobile phase. The presence of NH4Cl in the mobile phase induced the formation of [M+Cl] in the first quadrupole at m/z 397 and 409 for cortisol and 6alpha methylprednisolone (internal standard), respectively. In the collision cell, the complex dissociated to the neutral parent and the chloride ion at m/z 35; the latter ion was used for quantification. The calibration curve was linear from 0.5 to 100 ng/mL. The lower limit of quantification was 0.50 ng/mL and the limit of detection was 0.25 ng/mL. For quality control samples prepared in water, the intrabatch assay precision was 5.6%, 9.6%, and 9.9% at 50, 10, and 1 ng/mL, respectively. The interbatch assay precision was 4.2%, 6.3%, and 7.5% at 50, 10, and 1 ng/mL, respectively. For measurement of endogenous cortisol in plasma and urine samples, the intra-assay and interassay precision was 10.8% and 4.8% for total plasma cortisol, 13.1% and 5.2% for free plasma cortisol, 10.9% and 13.1% for cortisol protein-binding free fraction, and 8.9% and 14.4% for urine cortisol, respectively. A simple procedure of ultrafiltration coupled with the highly sensitive LC-MS/MS quantification offered a rapid and reproducible assay for plasma free cortisol, which may be useful in the assessment of adrenal function in patients, especially critically ill patients with abnormal protein binding. It may also be useful for plasma and urinary cortisol measurements in pharmacodynamic studies of adrenocorticoid response. PMID- 17417078 TI - 13C-erythromycin breath test as a noninvasive measure of CYP3A activity in newborn infants: a pilot study. AB - The intravenous C-erythromycin breath test (EBMT) has been largely used in adults as a validated probe to measure hepatic cytochrome P450 3A4 and 3A5 (CYP3A4/5) activity in vivo. Additionally, the oral EBMT has been suggested to measure combined hepatic and intestinal CYP3A4/5 activity. Both hepatic and intestinal CYP3A4/5 activities are low in neonates, but the exact developmental pattern is not known. Also, a knowledge gap exists on the impact of comedication or disease state on CYP3A4/5 activity in this population. However, to use the radioactive test in newborns is not feasible, for obvious ethical reasons. Hence, the aim of this pilot study was to determine if stable isotope-labeled C-erythromycin could be used alternatively. Preterm infants who needed treatment with erythromycin for ureaplasma infection were given an oral 10 to 15 mg/kg C-(N-dimethyl) erythromycin dose. Pharmacy regulations did not permit intravenous administration. Exhaled air samples were collected predose and up to 24 hours post-dose and analyzed for CO2 and CO2 with gas chromatography-mass spectrometry. Three patients received oral C-erythromycin. CO2 did not change significantly from baseline, showed a maximum blood concentration at 20 hours (+12%), and decreased over 24 hours (-16%) in these patients, respectively. Because none of these patients showed a consistent peak in C enrichment, in accordance with maximum blood concentration of oral erythromycin in preterms, we stopped this pilot trial after 3 patients. In conclusion, the lack of a consistent change in exhaled CO2 after oral C-erythromycin in this pilot study precludes the routine use of oral 13C-EBMT in preterm infants as a noninvasive probe of CYP3A4/5 activity. We speculate that this lack of change is due to developmentally low intestinal and hepatic CYP3A activity. PMID- 17417079 TI - A simple and sensitive high-performance liquid chromatographic method for quantification of PXD101, a histone deacetylase inhibitor in human plasma. AB - PXD101, a new histone deacetylase inhibitor, is currently undergoing phase I/II clinical trials as an anticancer drug. This study describes a simple and sensitive high-performance liquid chromatography ultraviolet method developed for the quantification of PXD101 in human plasma samples to support such trials. Following solid phase extraction at room temperature, the analytes were separated on a 5 mum C18 150 x 2.1 mm column using gradient elution (mobile phase of acetonitrile and 25 mM NaH2PO4, pH 2.8) at a flow rate of 0.5 mL/min and ultraviolet detection at 268 nm. Oxamflatin was used as an internal standard. PXD101 and the internal standard were eluted at about 7.9 min and 13.6 min, respectively. The lower limit of quantification of PXD101 in plasma was 10 ng/mL. The calibration curves for concentrations in the range of 10 to 2,000 ng/mL gave excellent linearity (r = 0.999). The coefficients of variation for intraday and interday assays were all less than 10%. The accuracy of all concentration determinations ranged from 98.0% to 102.0%. There were no problems with PXD101 stability following freeze-thawing, short-term exposure to room temperature, postextraction stability up to 24 hours, and sample storage at -70 degrees C for 3 months. The reported method, with dilution integrity of up to 50 fold, has the requisite sensitivity and is suitable for pharmacokinetic studies of PXD101. PMID- 17417080 TI - Doping-control analysis of the 5alpha-reductase inhibitor finasteride: determination of its influence on urinary steroid profiles and detection of its major urinary metabolite. AB - 5alpha-Reductase inhibitors such as finasteride are prohibited in sports according to the World Anti-Doping Agency. This class of drugs is used therapeutically to treat benign prostatic hyperplasia, as well as male baldness, by decreasing 5alpha-reductase activity. Accordingly, metabolic pathways of endogenous as well as synthetic steroids are influenced, which complicates the evaluation of steroid profiles in sports drug testing. The possibility of manipulating steroid excretion profiles and, presumably, to mask steroid abuse was investigated in 5 administration studies with use of finasteride at different doses, with and without coadministration of 19-norandrostenedione. The evaluation of urinary steroid profiles demonstrated the intense effect of finasteride on numerous crucial analytical parameters, in particular the production of 5alpha steroids such as androsterone and 5alpha-androstane-3alpha,17beta-diol, which was significantly reduced. In addition, the excretion of the main metabolite of norandrostenedione, norandrosterone, was significantly suppressed, by up to 84%, in elimination studies. For doping-control analysis the use of 5alpha-reductase inhibitors causes considerable problems because steroid profile parameters, which are commonly considered stable, are highly affected and complicate the detection of steroid abuse. In addition, the suppression of production and renal excretion of 5alpha-steroids such as 19-norandrosterone generated from anabolic agents such as 19-norandrostenedione may lead to false-negative doping-control results, because urine specimens are reported positive only when a threshold level of 2 ng/mL is exceeded. Finally, a method for the determination of the major urinary metabolite of finasteride (carboxy-finasteride) in routine doping-control screening with use of liquid chromatography-tandem mass spectrometry is described, allowing the detection of carboxy-finasteride for up to 94 hours in urine specimens collected after an oral administration of 5 mg of finasteride. PMID- 17417081 TI - Concentrations of scheduled prescription drugs in blood of impaired drivers: considerations for interpreting the results. AB - We report the concentrations of scheduled prescription drugs in blood samples from people arrested in Sweden for driving under the influence of drugs (DUID). The investigation covered a 2 year period 2004 (N = 7052 cases) and 2005 (N = 7759 cases) and was prompted by recent legislation stipulating zero-concentration limits in blood for controlled substances. However, prescription drugs are exempt from the zero-limit law provided that the medication was being used in accordance with a doctor's prescription. The blood concentrations of various psychoactive substances were compared with the limits of quantitation of the analytic method used and the so-called therapeutic concentration range according to various reference books and tabulations. Diazepam [N = 1950 (26%)] and nordazepam [N = 2168 (28%)] were the therapeutic agents most frequently identified in these forensic blood samples along with other benzodiazepines such as alprazolam [N = 430 (5.6%)], flunitrazepam [N = 308 (4.0%)], and nitrazepam [N = 222 (2.9%)]. The newer hypnotics, exemplified by zolpidem [N = 148 (1.9%)] and zopiclone [N = 111 (1.5%)], were also high on the list of psychoactive substances identified. Interpreting the concentration of a prescription drug in blood in relation to whether the person had taken an overdose or was abusing the substance in question is not always easy. The age, gender, degree of obesity, and ethnicity of the person concerned; the pharmacokinetic profile of the drug; polymorphism of drug metabolizing enzymes as well as liver and kidney function and blood hematocrit need to be considered. Among preanalytic factors, stability of the drug in blood after sampling, the type of tubes and preservatives used, the dosage form and route of administration deserve consideration. When therapeutic drug monitoring concentrations are compared with forensic toxicology results, then the plasma-to whole blood distribution ratio of the drug also needs to be considered. In blood samples from DUID suspects, the concentrations of many commonly used sedatives and hypnotics exceeded the accepted therapeutic limits, which gives an indication of the abuse potential of these types of medications. PMID- 17417082 TI - Estimation of the initial dose setting of vancomycin therapy with use of cystatin C as a new marker of renal function. AB - In recent years, it has been suggested that the glomerular filtration rate (GFR) can be predicted on the basis of serum cystatin C concentrations and that this measurement is more sensitive than serum creatinine concentration as a marker of renal function. In this study, to investigate the clinical utility of the initial dose setting of vancomycin by the population mean method with use of serum cystatin C as a marker of renal function, we compared the correlations between measured vancomycin concentrations and predicted vancomycin concentrations based on serum cystatin C or serum creatinine concentrations in elderly (>/=65 years old) and nonelderly (<65 years old) patients. An analysis of prediction accuracy (bias) and precision was evaluated by calculating the mean prediction error (ME), the mean absolute error (MAE), and the root mean squared prediction error (RMSE). For nonelderly patients (n = 50), there was no significant difference in the MAE based on the use of serum creatinine or serum cystatin C concentration. However, for elderly patients (n = 105), the MAE based on serum cystatin C concentration was significantly better than that based on serum creatinine level. These results suggest that serum cystatin C is a good marker of renal function in comparison with serum creatinine for dose setting of vancomycin, especially in an elderly population. PMID- 17417085 TI - Magnetic resonance imaging of uterine malignancies. AB - Magnetic Resonance Imaging (MRI) is the criterion standard in the assessment and staging of uterine cancer because of the high tissue contrast between glandular lining and inner and outer muscle layers of the uterine corpus and cervix on T2 weighted scans. It is also an essential tool in monitoring treatment response and in assessing disease recurrence in these patients. The key to a successful examination lies in good patient preparation, use of antiperistaltic agents, and a meticulous scanning technique. Endometrial carcinomas are the most common uterine malignancy, occurring in a primarily postmenopausal population. Dynamic contrast-enhanced scans may be required in addition to T2-weighted images to assess the presence of myometrial invasion in this age group. Cervical cancers occur in a younger population and are readily assessed with T2-weighted imaging. Use of an endovaginal receiver coil allows high spatial resolution imaging of the cervix, which is particularly useful when assessing patients for fertility sparing procedures such as trachelectomy. Uterine sarcomas are 1% to 3% of all uterine malignancies and usually arise from a mixture of homologous and heterologous elements. Rarely, lymphoma, small cell carcinoma and metastatic deposits involve the uterus. This review summarizes the optimal scanning techniques for demonstrating uterine malignancy and discusses the role of imaging and the MRI appearances of uterine tumors. PMID- 17417086 TI - Magnetic resonance imaging of adnexal masses. AB - Adnexal masses are common in women of all ages. A range of physiological and benign ovarian conditions that develop in women, especially in the reproductive age, and adnexal malignancies can be evaluated with magnetic resonance imaging (MRI). Management of women with adnexal masses is frequently guided by imaging findings; therefore, precise characterization of adnexal pathology should be performed whenever possible. Magnetic resonance imaging is useful in characterization of adnexal masses that are not completely evaluated by ultrasound because it can provide additional information on soft tissue composition of adnexal masses based on specific tissue relaxation times and allows multiplanar imaging at large field of view to define the origin and extent of pelvic pathology. The patients most likely to benefit from MRI are pregnant women and those who are premenopausal and have masses that have complex features on ultrasound but do not have raised cancer antigen 125 tumor marker levels. The overlap in imaging appearance among different cell type malignancies makes it difficult to predict exact histology based on MRI appearance; however, MRI has a high accuracy in differentiating benign from malignant masses. Teratomas, endometriomas, simple and hemorrhagic cysts, fibromas, exophytic or extrauterine fibroids, and hydrosalpinges can be diagnosed with high confidence. In this article, the authors review the histopathologic background and MRI features of adnexal masses and discuss the role of MRI in the differentiation of benign from malignant adnexal pathologies. PMID- 17417087 TI - Magnetic resonance imaging of benign uterine pathology. AB - Magnetic resonance imaging is commonly used for the identification and characterization of many pelvic abnormalities. Magnetic resonance provides the most comprehensive and detailed view of the uterus of any imaging modality. This article focuses on the magnetic resonance imaging features used to recognize and describe congenital uterine anomalies and benign conditions of the uterus. PMID- 17417088 TI - Magnetic resonance imaging of abdominal pain during pregnancy. AB - The objective of this review is to demonstrate magnetic resonance imaging as an important adjunct to ultrasound and computed tomography in the evaluation of the pregnant patient with abdominal pain. With the advent of ultrafast T2-weighted pulse sequences, fetal and bowel motion cause minimal artifact on the images. An accurate diagnosis can often be made in a few minutes based on these high contrast images performed in 2 or 3 planes. T1-weighted gradient echo images with and without fat saturation are useful for identifying blood and fat, especially in the case of adnexal masses. Gadolinium-diethylenetriamine pentaacetic acid is rarely used to diagnose inflammatory or obstructive disease and is reserved for those patients with suspected malignancies. PMID- 17417089 TI - Magnetic resonance imaging of pelvic floor defects in women. AB - Contemporary fast magnetic resonance imaging techniques allow dynamic evaluation of the entire female pelvic floor with excellent visualization of pelvic organs and muscular and fascial supportive structures in a single noninvasive study that does not expose the patient to ionizing radiation. This article focuses on the role of magnetic resonance imaging in defining pelvic floor defects that can guide surgical management of women with pelvic organ prolapse, especially those who undergo evaluation for symptoms of multicompartmental involvement before a complex pelvic floor reconstruction or those who have failed previous repairs. PMID- 17417090 TI - Imaging the female pelvis at 3.0 T. AB - Three-Tesla whole body imaging is rapidly becoming part of routine clinical practice. Although it is generally thought that pelvic imaging at 3.0 T will be beneficial because of increased signal to noise and greater spectral separation, adjustments in protocol and sequence parameters are necessary to optimize image quality. The question remains as to whether 3.0-T imaging will offer further benefits beyond 1.5 T in terms of lesion characterization and functional imaging. This article aims to address safety concerns and to illustrate the potential benefits and technical challenges of imaging the female pelvis at 3.0 T. Imaging protocols and sequence parameters for routine gynecologic indications are suggested, and potential clinical applications at 3.0 T are discussed such as magnetic resonance spectroscopy, perfusion, diffusion weighted imaging, and the use of alternate contrast agents. PMID- 17417091 TI - Case report: Fatigue fracture of the proximal fibula with secondary common peroneal nerve injury. AB - Stress fractures of the proximal fibula are uncommon and usually result from axial loading, which is mostly described in runners. We report an unusual mechanism of such a fracture in a circus performer resulting from repetitive direct horizontal loading from a trapeze bar. In addition, the bony injury resulted in a secondary injury to the common peroneal nerve with corresponding weakness. Both injuries responded well to nonoperative treatment and the athlete had an excellent recovery with no residual symptoms. He was able to resume his training with the use of protective padding applied to the proximal legs. Fracture of the proximal fibula caused by direct repetitive stress to the bone with a secondary compression injury to the common peroneal nerve is a previously undescribed injury. We report a patient who presented with this injury, the possible mechanisms of such injury, its management, and outcome. PMID- 17417092 TI - Absence of HOXD10 mutations in idiopathic clubfoot and sporadic vertical talus. AB - The genetic etiology of idiopathic clubfoot is unknown. There have been cases reported in which both clubfoot and vertical talus appears in the same family; therefore, the genes responsible for vertical talus are reasonable candidates for idiopathic clubfoot. A mutation in HOXD10 was previously identified in a family with isolated congenital vertical talus. To determine whether HOXD10 is involved in the etiology of idiopathic clubfoot, HOXD10 coding and 5' and 3' untranslated regions were resequenced in 190 patients (177 with clubfoot, 10 with sporadic vertical talus, and 3 with both clubfoot and vertical talus), and 160 ethnically matched control subjects. Rare nonsynonymous HOXD10 amino acid substitutions (Leu154Val, Asn202Lys, and Thr175Ala), likely benign variants, were all detected once in patients and control subjects. Nucleotide substitutions were also identified in HOXD10 intronic and 3' untranslated regions, but were not more frequent in cases compared to controls. To investigate the possibility that unsequenced regulatory regions play a role in this disorder, we performed linkage analysis with markers on chromosome 2q near HOXD10 in one large family. We found no evidence of linkage near the HOXD gene cluster on chromosome 2q, suggesting genes other than HOXD10 are responsible for idiopathic clubfoot. PMID- 17417093 TI - Joint salvage for pathologic fracture of giant cell tumor of the lower extremity. AB - Pathologic fracture through giant cell tumor is thought to be associated with higher rates of recurrence and poor functional outcome. We compared patients with and without pathologic fracture through giant cell tumor of weightbearing long bones. We retrospectively reviewed 139 patients with giant cell tumor of weightbearing long bones with (n = 43) and without (n = 96) pathologic fracture at presentation; the two groups had similar demographics. Joint salvage was successful in 84% of the fracture group and 96% of the nonfracture group. Five year recurrence-free survival rates were comparable between the two groups (82.6% [95% confidence interval, 69.1-95.9%] in the fracture group and 77.9% [95% confidence interval, 67.7-88.1%] in the non-fracture group). There was a trend toward lower 5-year metastatic-free survival in the fracture group (94.7% [95% confidence interval, 87.3-100%]) than in the nonfracture group (97.3% [95% confidence interval, 93.5-100%]). Functional outcome was good and similar in the two groups. Arthrofibrosis was more common in the group with pathologic fracture. Joint salvage for patients with pathologic fractures through giant cell tumor of weightbearing bones is a reasonable option with functional outcomes and recurrence rates comparable to those of patients without fracture. PMID- 17417094 TI - Functional outcomes after treatment of aggressive tumors in the distal radius. AB - We aimed to determine whether wide excision and radiocarpal arthrodesis in patients with aggressive tumors of the distal radius resulted in more significant functional deficit than joint salvage procedures. We compared functional outcomes, surgical complications, and local tumor relapse in 33 consecutive patients. Fourteen patients initially had joint-sparing procedures, whereas 19 underwent wide resection and arthrodesis. At minimum final followup of 18 months (mean, 96 months; range, 18-204 months), local tumor relapse occurred in five of 14 patients after joint salvage versus zero of 19 after arthrodesis. Two of five patients with tumor recurrence retained their native joints, whereas three underwent resection and arthrodesis. Six of 19 patients had complications, including one nonunion, for which we performed additional surgery after arthrodesis. All patients remained disease-free at followup and none developed metastases. Disabilities of the Arm, Shoulder and Hand score, Toronto Extremity Salvage Score, and Musculoskeletal Tumor Society scores all indicated more disability after arthrodesis. Persistent pain, radiographic wrist arthritis, and weaker grip strength was seen in some patients following joint salvage. Whenever possible, joint preservation is preferable for the result of better extremity function, but weaker grip, higher local tumor relapse, and late arthritis remain concerns. PMID- 17417095 TI - Large tumors of the axilla: limb-sparing resection versus amputation in 27 patients. AB - Tumors of the axilla are rare and pose a surgical challenge because they are usually large at presentation and in close proximity to the major neurovascular bundle of the upper extremity. The use of detailed preoperative evaluation studies and extensile surgical exposure for these tumors enabled us to determine tumor resectability and proceed with a safe resection or perform an amputation when required. We retrospectively reviewed 27 patients who underwent resection of an axillary tumor from 1989 to 2004 and analyzed their presenting symptoms, results of preoperative studies, type of surgery, and functional outcome. Tumors were exposed using a utilitarian shoulder approach that revealed no tumor invasion of the neurovascular bundle in 19 patients and invasion in eight. The former group was treated with tumor resection and the latter with forequarter amputation. Neurologic deficit, limb edema, and angiographic observation of arterial narrowing were associated with amputation. Good function was achieved in 15 of 19, shoulder range of motion was preserved in 14 of 19, and local tumor control was maintained in 16 of 19 patients who underwent a limb-sparing resection. PMID- 17417097 TI - Infected distal biceps tendon repairs: three case reports. AB - Numerous reports address the risk of infection with surgery about the elbow. However, there is little information regarding treatment of infected distal biceps repairs. We reviewed the records of 37 consecutive patients with biceps repairs between January 1, 2000, and January 1, 2005, to identify cases of postoperative infection. Three patients were identified and their records examined to identify the responsible organisms and any forms of treatment. Two patients presented acutely less than 4 weeks after surgery with frank purulence draining from the incision area. The third patient presented 4 months after tendon repair. In both patients with acute infection, the infection resolved without removal of any stitches or hardware. In the patient with chronic infection, the anchoring material was removed because it was no longer holding the biceps tendon to the radial tuberosity. These data suggest acute infections can be treated with antibiotics and debridement without taking down the tendon repair. However, as with any chronic musculoskeletal infection, all foreign material may need removal before a second repair. Our data demonstrate deep infection is not an uncommon complication of distal biceps tendon repairs that initially can be treated without taking down the construct. PMID- 17417098 TI - Limb salvage surgery for osteosarcoma: effective low-cost treatment. AB - Successful management of osteosarcoma with limb salvage surgery is a challenging problem in the developing world. We report our early results with low-cost chemotherapy (without methotrexate) and low-cost limb salvage surgery. We prospectively collected data for 135 patients with histologically proven high grade osteosarcoma of the extremities given neoadjuvant chemotherapy and treated with limb salvage surgery between January 2000 and February 2004. A locally designed and fabricated stainless steel customized megaprosthesis was used in 78 patients. Disease status and function was evaluated at followup ranging from 24 to 72 months. Followup data were available for 120 of the 135 patients. There were eight infections, four prosthesis breakages and three loosenings. Eighteen patients had local recurrence; 17 of these patients also developed lung metastases. Seventy-three patients (61%) were disease-free at followup. The group of 34 patients with 100% necrosis had better disease-free survival (79%). According to a modified Enneking system, the average functional score was 25.5 of 30 (85%) for the lower extremity and 20 of 30 (66%) for the shoulder. Our preliminary results suggest osteosarcoma can be managed well in a developing country in a cost-effective way. Limb salvage surgery has now become the standard of care. PMID- 17417099 TI - Simple triage scoring system predicting death and the need for critical care resources for use during epidemics. AB - OBJECTIVES: In the event of pandemic influenza, the number of critically ill victims will likely overwhelm critical care capacity. To date, no standardized method for allocating scarce resources when the number of patients in need far exceeds capacity exists. We sought to derive and validate such a triage scheme. DESIGN: : Retrospective analysis of prospectively collected data. SETTING: Emergency departments of two urban tertiary care hospitals. PATIENTS: Three separate cohorts of emergency department patients with suspected infection, comprising a total of 5,133 patients. INTERVENTIONS: None. MEASUREMENTS: A triage decision rule for use in an epidemic was developed using only those vital signs and patient characteristics that were readily available at initial presentation to the emergency department. The triage schema was derived from a cohort at center 1, validated on a second cohort from center 1, and then validated on a third cohort of patients from center 2. The primary outcome for the analysis was in-hospital mortality. Secondary outcomes were intensive care unit admission and use of mechanical ventilation. MAIN RESULTS: Multiple logistic regression demonstrated the following as independent predictors of death: a) age of >65 yrs, b) altered mental status, c) respiratory rate of >30 breaths/min, d) low oxygen saturation, and e) shock index of >1 (heart rate > blood pressure). This model had an area under the receiver operating characteristic curve of 0.80 in the derivation set and 0.74 and 0.76 in the validation sets. When converted to a simple rule assigning 1 point per covariate, the discrimination of the model remained essentially unchanged. The model was equally effective at predicting need for intensive care unit admission and mechanical ventilation. CONCLUSIONS: If, as expected, patient demand far exceeds the capability to provide critical care services in an epidemic, a fair and just system to allocate limited resources will be essential. The triage rule we have developed can serve as an initial guide for such a process. PMID- 17417100 TI - High prevalence of subtherapeutic plasma concentrations of efavirenz in children. AB - OBJECTIVE: Efavirenz-containing regimens using concentration-controlled dosing have been shown to provide potent antiretroviral activity in children. In many settings, concentration-controlled dosing is not available. In this study, efavirenz plasma concentrations were evaluated in South African HIV-infected children receiving efavirenz-based antiretroviral treatment. METHODS: Three consecutive blood samples were drawn between 12 and 24 hours after dosing in 15 HIV-infected children receiving the recommended daily doses of efavirenz. Validated liquid chromatography tandem mass spectrometry methods were used to determine plasma levels of efavirenz. The trough concentration (Cmin) of efavirenz was estimated by extrapolation of the log-linear regression line of the 3 concentration versus time points to 24 hours. RESULTS: The estimated Cmin was <1 mg/L in 6 (40%) of the children. Three of the 5 children with detectable viral loads had low efavirenz concentrations. Marked bimodality in efavirenz concentrations was observed. CONCLUSIONS: Our findings, together with those of previous studies, indicate that many children dosed according to the current guidelines do not achieve adequate efavirenz exposure. Because low efavirenz concentrations are associated with the rapid emergence of efavirenz-resistant mutations and treatment failure, the current recommended efavirenz doses should be re-evaluated, especially in developing countries, where therapeutic drug monitoring is seldom available. PMID- 17417101 TI - HIV-1 Subtype C gag-specific T-cell responses in relation to human leukocyte antigens in a diverse population of HIV-infected Ethiopians. AB - Knowledge of the most dominant T-cell epitopes in the context of the local human leukocyte antigen (HLA) background is a prerequisite for the development of an effective HIV vaccine. In 100 Ethiopian subjects, 16 different HLA-A, 23 HLA-B, and 12 HLA-C specificities were observed. Ninety-four percent of the population carried at least 1 of the 5 most common HLA-A and/or HLA-B specificities. HIV specific T-cell responses were measured in 48 HIV-infected Ethiopian subjects representing a wide range of ethnicities in Ethiopia using the interferon (IFN) gamma enzyme-linked immunospot (Elispot) assay and 49 clade C-specific synthetic Gag peptides. Fifty-eight percent of the HIV-positive study subjects showed T cell responses directed to 1 or more HIV Gag peptides. Most Gag-specific responses were directed against the subset of peptides spanning Gag p24. The breadth of response ranged from 1 to 9 peptides, with most (78%) individuals showing detectable responses to <3 Gag peptides. The magnitude of HIV-specific T cell responses was not associated with HIV viral load but correlated positively with CD4 T-cell counts. The most frequently targeted Gag peptides overlapped with those previously described for HIV-1 subtype C-infected southern Africans, and therefore can be used in a multiethnic vaccine. PMID- 17417102 TI - HIV prevalence and undiagnosed infection among a community sample of gay men in Scotland. AB - OBJECTIVE: To examine HIV prevalence among men in gay bars in Glasgow and Edinburgh, Scotland. METHODS: Self-complete questionnaires and Orasure oral fluid collection kits (OraSure Technologies, Inc., Bethlehem, PA), to collect samples to be tested anonymously for HIV antibodies, were distributed. A total of 1744 men completed questionnaires (66.1% response rate), and 1350 provided samples (51.6% response rate). RESULTS: HIV prevalence was 4.4% (95% confidence interval: 3.5% to 5.7%). Positivity was associated with older age (mean of 36 years for positive men vs. 32 years for negative men), having 10 or more anal intercourse (AI) partners (12.8% positive vs. 3.4% of men with <10 AI partners, P < 0.05), and self-reported sexually transmitted infections (STIs) in the previous year (12.3% positive vs. 3.5% of men without STIs in the previous year; P < 0.05). Of the HIV-positive men, 41.7% were undiagnosed. More than half had had a negative HIV test result and perceived themselves to be HIV-negative. Men who had not used a genitourinary medicine clinic or had an STI in the previous year were more likely to be undiagnosed (65.0% and 52.5%, respectively). CONCLUSIONS: This is the first study to assess HIV prevalence among a community sample of gay men in Scotland. There is an urgent need for HIV prevention efforts to address the high level of undiagnosed infection and incorrect assumptions of status. PMID- 17417103 TI - Probability sample estimates of bathhouse sexual risk behavior. AB - OBJECTIVE: Previous research links high rates of unprotected anal intercourse (UAI) with men who go to bathhouses; however, the literature provides no prevalence estimates. An exit survey of a probability sample was conducted to describe the prevalence of risk activity at the bathhouse. METHODS: Data are from a 2-stage probability sample of men exiting a gay bathhouse (n = 400). RESULTS: During their visit, 91.5% of men had oral sex and 44.2% had anal sex (11.1% reported UAI and 5.5% reported unprotected receptive anal intercourse). In the prior 3 months, 85% reported having anal sex, which was more likely to be unprotected when it occurred in a private home or hotel as opposed to a public setting (P < 0.001). Moreover, having UAI at home was a significant correlate of risk during the bathhouse visit (P < 0.001). CONCLUSIONS: Most men at the bathhouse engaged in oral sex rather than anal sex, and most anal sex included use of condoms. Furthermore, men were more likely to have UAI in a private home than in any public setting. The bathhouse seems to have facilitated condom use when anal sex occurred on-site. PMID- 17417104 TI - Hunsaker Mon-Jet tube for secondary tracheoesophageal puncture. PMID- 17417105 TI - Emerging trends in the performance of parathyroid surgery. AB - OBJECTIVE: The management of hyperparathyroidism has evolved rapidly in the past decade with the introduction of intraoperative parathyroid hormone testing, radioguided surgery, and endoscopic surgery. Not surprisingly, there is a corresponding movement toward specialization of surgeons providing increasingly sophisticated treatments for head and neck endocrine disorders. We sought to identify trends in the disciplines performing parathyroid surgery. DESIGN: Nonrandomized, controlled comparison of surgical caseloads and publication volumes. METHODS AND MATERIALS: Two metrics designed to reflect the proportion of parathyroidectomies being performed by otolaryngologist-head and neck surgeons (OHNS) and general surgeons (GS) were chosen: the operative case logs of graduates from American training programs in OHNS and GS from 1996 through 2005 and the number of scientific papers published relating to parathyroid surgery during two timeframes (1991-1995 and 2001-2005). RESULTS: There was a gradual increase in the mean number of parathyroid surgeries performed by GS residents from 6.0 in 1996 to a peak of 9.2 in 2004; this volume has begun to decline (to 8.5 in 2005). During the same timeframe, the mean number of parathyroidectomies performed by OHNS residents rose sharply and steadily from 1.8 in 1996 to 10.9 in 2005. The number of American GS parathyroid publications from 1991 to 1995 was 41, compared with 108 in the period 2001 to 2005. During the same timeframe, the number of American OHNS parathyroid papers increased from 1 to 27. The relative proportion of parathyroid publications authored by American otolaryngologists rose from 2.4% to 20.0% (P = .006). CONCLUSIONS: Increasingly, otolaryngologists are the primary surgeons in parathyroid operations as indicated by two surrogate metrics. Graduating chief residents in otolaryngology now perform more parathyroid procedures than chief residents in general surgery, and a growing proportion of parathyroid publications are being authored by otolaryngologists. PMID- 17417106 TI - Transpterygoid transposition of a temporoparietal fascia flap: a new method for skull base reconstruction after endoscopic expanded endonasal approaches. AB - BACKGROUND: Endoscopic expanded endonasal approaches (EEAs) for the resection of lesions of the anterior and ventral skull base can create large defects that present a significant risk of postoperative cerebrospinal fluid (CSF) leak. These defects, especially in patients who received preoperative radiotherapy, are best reconstructed with vascularized tissue. The Hadad-Bassagasteguy flap, a pedicled nasoseptal flap, is our preferred method for reconstruction. This option is not available, however, in patients who underwent a previous posterior septectomy or in those with tumors that invade the pterygopalatine fossa (PPF) or sphenoid sinus rostrum. In this scenario, we have used a temporoparietal fascial flap (TPFF) for the reconstruction of large surgical defects. METHODS: We developed a new technique for the transposition of the TPFF into the nasal cavity to reconstruct skull base defects after EEA. The flap is harvested using a conventional hemicoronal incision but is then advanced to the defect using a temporal-infratemporal tunnel and an endonasal transpterygoid approach. The latter is created using an endoscopic endonasal approach that involves the resection of the posterior wall of the antrum, dissection of the PPF, and partial resection of the pterygoid plates. These maneuvers open a bone window to accommodate the flap. The soft tissue tunnel, extending from the temporal to the infratemporal and then to the PPF, is opened with percutaneous tracheostomy dilators. We present a detailed description of the surgical technique and a retrospective review of two cases in which we used this technique. RESULTS: Two patients with large CSF fistulas who had undergone preoperative radiotherapy were reconstructed transposing the TPFF through a transpterygoid tunnel. We obtained an adequate exposure for placing the flap endonasally, and the flap provided complete coverage of the skull base defect. Both CSF leaks were resolved without any additional morbidity from the flap or the access technique. CONCLUSION: The TPFF is a reliable and versatile method for the reconstruction of the anterior, middle, clival, and parasellar skull base after EEAs. Its harvesting requires an external incision; thus, it is not our preferred method of reconstruction. It is recommended for large dural defects in patients with previous posterior septectomy and previous radiation treatment. PMID- 17417107 TI - Utility of lip biopsy in the diagnosis and treatment of Sjogren's syndrome. AB - OBJECTIVE: Sjogren's syndrome (SS) is an autoimmune disease involving primarily the salivary and lacrimal glands ("sicca" symptoms) with extraglandular features including joint and nervous system involvement. In patients with a questionable diagnosis of SS but severe extraglandular symptoms, a lip biopsy is often performed to firmly establish the diagnosis of SS. This study was undertaken to identify areas of uncertainty regarding the accuracy of biopsy interpretation and the diagnostic benefits of the procedure. STUDY DESIGN: : Retrospective review of clinical and pathologic data. METHODS: Clinical data from 47 patients referred to the otolaryngology-head and neck surgery service for lip biopsy were reviewed. Archival pathologic specimens were scored using the currently accepted grading system. RESULTS: The grading system was incorrectly applied during initial interpretation of 45% (n = 21) of specimens. This resulted in five (10%) pathologic misdiagnoses and 16 (34%) nondiagnoses. On re-interpreting the specimens with consistent application of the grading system, 62% (n = 29) of the biopsies were definitely positive, and 36% (n = 17) were negative. Neither positive serology nor the presence of sicca symptoms predicted a positive biopsy (likelihood ratio = 0.95 and 0.96, respectively). Lip biopsy guided treatment in at least 65% (n = 31) of patients but was ignored in the presence of other clinical findings in 17% (n = 8) of patients. CONCLUSIONS: Consistent application of the grading system is essential in avoiding incorrect diagnosis and aiding clinical decisions. However, not all patients undergoing lip biopsy will derive diagnostic benefit from the procedure. In this series, clinical symptoms and serology did not predict positive lip biopsy. PMID- 17417108 TI - Functional outcome of vocal fold medialization thyroplasty with a hydroxyapatite implant. AB - OBJECTIVES: Unilateral vocal fold paralysis can cause a persistent incomplete glottal closure during phonation, resulting in impaired voice function. The aim of this study was to evaluate functional results of medialization thyroplasty using a hydroxyapatite implant (VoCoM). STUDY DESIGN: Prospective observational cohort study. METHODS: Between 1999 and 2003, a total of 26 patients (19 men, 7 women) undergoing medialization thyroplasty using a hydroxyapatite implant because of unilateral vocal fold paralysis were enrolled in the study. To evaluate voice function, the following parameters were measured preoperatively and postoperatively: mean fundamental frequency, mean sound pressure level, frequency and amplitude range (voice range profile), and maximum phonation time. A perceptual assessment of hoarseness was conducted using the Roughness, Breathiness, Hoarseness scale. Furthermore, the magnitude of voice related impairment of the patient's communication skills was rated on a 7-point scale. A combined parameter called the Voice Dysfunction Index (VDI) was used to rate vocal performance. RESULTS: All patients showed a statistically significant improvement in the VDI, in perceptual voice analysis, in maximum phonation time, and in the dynamic range of voice. One patient experienced a postoperative wound hemorrhage as a minor complication. No further complications or implant extrusions were observed. CONCLUSIONS: Medialization thyroplasty using a hydroxyapatite implant is a secure and efficient phonosurgical procedure. Voice quality and patient satisfaction improve significantly after treatment. PMID- 17417109 TI - Activity/stability of human pepsin: implications for reflux attributed laryngeal disease. AB - OBJECTIVES/HYPOTHESIS: Exposure of laryngeal epithelia to pepsin during extra esophageal reflux causes depletion of laryngeal protective proteins, carbonic anhydrase isoenzyme III (CAIII), and squamous epithelial stress protein Sep70. The first objective of this study was to determine whether pepsin has to be enzymatically active to deplete these proteins. The second objective was to investigate the effect of pH on the activity and stability of human pepsin 3b under conditions that might be found in the human esophagus and larynx. STUDY DESIGN: Prospective translational research study. METHODS: An established porcine in vitro model was used to examine the effect of active/inactive pepsin on laryngeal CAIII and Sep70 protein levels. The activity and stability of pepsin was determined by kinetic assay, measuring the rate of hydrolysis of a synthetic pepsin-specific substrate after incubation at various pH values for increasing duration. RESULTS: Active pepsin is required to deplete laryngeal CAIII and Sep70. Pepsin has maximum activity at pH 2.0 and is inactive at pH 6.5 or higher. Although pepsin is inactive at pH 6.5 and above, it remains stable until pH 8.0 and can be reactivated when the pH is reduced. Pepsin is stable for at least 24 hours at pH 7.0, 37 degrees C and retains 79% +/- 11% of its original activity after re-acidification at pH 3.0. CONCLUSIONS: Detectable levels of pepsin remain in laryngeal epithelia after a reflux event. Pepsin bound there would be enzymatically inactive because the mean pH of the laryngopharynx is pH 6.8. Significantly, pepsin could remain in a form that would be reactivated by a subsequent decrease in pH, such as would occur during an acidic reflux event or possibly after uptake into intracellular compartments of lower pH. PMID- 17417111 TI - Barotrauma and decompression illness of the inner ear: 46 cases during treatment and follow-up. AB - INTRODUCTION: Diving accidents affecting the inner ear are much more common than was once thought. Among the 319 patients treated in our clinic between January 2002 and November 2005, 46 cases involved 44 divers with symptoms of acute inner ear disorders. The objective of the present article is to investigate the symptoms of the acute disorders and assess any residual damage. STUDY DESIGN: Retrospective case analysis. MATERIALS AND METHODS: The medical records were used to study the cases of 18 divers treated for inner ear decompression illness on 20 occasions and 26 divers who had inner ear barotrauma. The symptoms of the disorder at the beginning of treatment, latency period before the first therapeutic measures, kind of initial therapy, symptoms after the accident, and hearing and balance functions at the last examination in our clinic were assessed. Divers with inner ear decompression illness were examined via means of transcranial or carotid Doppler ultrasonography for the presence of a vascular right-to-left (R/L) shunt. RESULTS: Of 18 divers with inner ear decompression illness, 17 reported vertigo as the main symptom. In one diver, the inner ear decompression illness was manifested bilaterally. The divers with inner ear decompression illness had been treated with hyperbaric oxygen therapy in 14 of 20 cases; the average latency period before the start of therapy was 40 hours (median, 10 h). In 15 (83%) of 18 patients, a large R/L shunt was detected, and in 14 (78%) of 18 patients, residual cochleovestibular damage was detected. Only 9 of 26 patients with inner ear barotrauma mentioned feeling dizzy, and in no patient was vertigo the main symptom. Twenty-one patients complained of tinnitus, whereas 20 complained of hearing loss. The hearing loss ranged from an unobtrusive difference of 10 dB between the ears up to complete deafness. Three patients were subjected to tympanoscopy because of suspected rupture of the round window membrane. Of patients with inner ear barotrauma, 78% had residual cochleovestibular damage. CONCLUSION: We describe for the first time a patient with bilateral manifestation of inner ear decompression illness. Inner ear decompression illness is frequently associated with a R/L shunt; therefore, after a diving accident, the patient's fitness to dive should be assessed via a specialist in diving medicine. Both decompression illness and barotrauma of the inner ear result in residual cochleovestibular damage in more than three of four patients. PMID- 17417113 TI - Implementation of a medical emergency team in a large pediatric teaching hospital prevents respiratory and cardiopulmonary arrests outside the intensive care unit. AB - OBJECTIVE: We implemented a medical emergency team (MET) in our free-standing children's hospital. The specific aim was to reduce the rate of codes (respiratory and cardiopulmonary arrests) outside the intensive care units by 50% for >6 months following MET implementation. DESIGN: Retrospective chart review and program implementation. SETTING: A children's hospital. PATIENTS: None. INTERVENTIONS: The records of patients who required cardiorespiratory resuscitation outside the critical care areas were reviewed before MET implementation to determine activation criteria for the MET. Codes were prospectively defined as respiratory arrests or cardiopulmonary arrests. MET preventable codes were prospectively defined. The incidence of codes before and after MET implementation was recorded. MEASUREMENTS AND MAIN RESULTS: Twenty-five codes occurred during the pre-MET baseline compared with six following MET implementation. The code rate (respiratory arrests + cardiopulmonary arrests) post-MET was 0.11 per 1,000 patient days compared with baseline of 0.27 (risk ratio, 0.42; 95% confidence interval, 0-0.89; p = .03). The code rate per 1,000 admissions decreased from 1.54 (baseline) to 0.62 (post-MET) (risk ratio, 0.41; 95% confidence interval, 0-0.86; p = .02). For MET-preventable codes, the code rate post-MET was 0.04 per 1,000 patient days compared with a baseline of 0.14 (risk ratio, 0.27; 95% confidence interval, 0-0.94; p = .04). There was no difference in the incidence of cardiopulmonary arrests before and after MET. For codes outside the intensive care unit, the pre-MET mortality rate was 0.12 per 1,000 days compared with 0.06 post-MET (risk ratio, 0.48; 95% confidence interval, 0-1.4, p = .13). The overall mortality rate for outside the intensive care unit codes was 42% (15 of 36 patients). CONCLUSIONS: Implementation of a MET is associated with a reduction in the risk of respiratory and cardiopulmonary arrest outside of critical care areas in a large tertiary children's hospital. PMID- 17417114 TI - Increasing use of extracorporeal life support in methicillin-resistant Staphylococcus aureus sepsis in children. AB - BACKGROUND: Pediatric cases of fulminant community-associated methicillin resistant Staphylococcus aureus (MRSA) infections requiring extracorporeal life support (ECLS) have been reported, but the frequency of ECLS use for severe presentations of staphylococcal disease is unknown. OBJECTIVE: To describe the frequency and characteristics of children with MRSA infections requiring ECLS using local and international databases. METHODS: The reasons for use of ECLS in children 0-18 yrs of age were determined in both the Vanderbilt Children's Hospital medical record system and the Extracorporeal Life Support Organization database during the years 1994-2005. Demographic characteristics, ventilatory management, and measurements of cardiopulmonary status in subjects undergoing ECLS with a pre-ECLS diagnosis of infection with Staphylococcus aureus and MRSA were included. RESULTS: Three subjects with MRSA sepsis requiring ECLS were identified at Vanderbilt since 2000. Before that time, no cases due to MRSA were reported. The three subjects were previously healthy adolescents with severe necrotizing pneumonia associated with skin/soft-tissue infection and two died. A total of 45 patients requiring ECLS for MRSA infection were identified in the International Extracorporeal Life Support Organization database, with nearly half reported in the past 2 yrs (20 of 45 patients). The median age was 2.4 yrs (interquartile range, 0.36-14 yrs), with peaks noted in infancy and adolescence. In Extracorporeal Life Support Organization subjects with MRSA, survival to discharge was highest in infants and young children aged 1-4 yrs (65% and 71%, respectively) and lowest in the age ranges of 5-9 yrs and 13-18 yrs (0% and 31%, respectively). There were no statistically significant differences in pre-ECLS ventilatory settings, cardiopulmonary status, or frequency of complications between survivors and nonsurvivors. CONCLUSIONS: The use of ECLS for MRSA infection seems to be increasing both locally and internationally. High mortality rates, particularly in older patients, are concerning and highlight the increasing problem with this pathogen. PMID- 17417115 TI - Pertussis: severe clinical presentation in pediatric intensive care and its relation to outcome. AB - OBJECTIVE: To describe our institutional experience in the management of infants and children with pertussis admitted during a 20-yr period (January 1985 through December 2004) and also to study the relation between method of presentation and outcome. SETTING: Pediatric intensive care unit in a university-affiliated tertiary pediatric hospital in Melbourne, Australia. DESIGN/METHODS: Retrospective review of medical records and radiology reports of patients with a diagnosis of pertussis identified from the pediatric intensive care unit database. RESULTS: A total of 49 patients (median age, 6 wks; interquartile range, 4-8 wks) required 55 admission episodes to the pediatric intensive care unit. Main reasons for admission were apnea with or without cough paroxysms (63%), pneumonia (18%), and seizures (10%). None of the infants had completed the primary course of immunization, and 94% had not received a single dose of pertussis vaccine. Infants presenting with pneumonia presented earlier (p = .001), had longer intensive care stay (p = .007), higher white cell count (p < or = .001), lower Pao2 at admission (p = .020), and higher mortality. Six infants out of seven needing circulatory support died (including all four treated with extracorporeal membrane oxygenation), and all deaths (n = 7) occurred in infants who had pneumonia at presentation. CONCLUSION: Patients with pertussis, presenting as apnea (with or without cough paroxysms), treated in the pediatric intensive care unit had 100% survival. However, pneumonia as the main reason for admission and the need for circulatory support is associated with a very poor outcome. A deeper understanding of the molecular basis of Bordetella pertussis and its relation to the human host might offer means for future therapies. PMID- 17417116 TI - A critical appraisal of "Blind and bronchoscopic sampling methods in suspected ventilator-associated pneumonia: a multicentre prospective study" by Mentec et al. (Intensive Care Med 2004; 30:1319-1326). PMID- 17417117 TI - Energy expenditure in critically ill children. AB - OBJECTIVE: To assess the measured resting energy expenditure pattern over time in a group of critically ill children who were admitted to a pediatric intensive care unit and to determine whether a hypermetabolic response, i.e., >10% above predicted, occurred in a pattern similar to that observed in adults. A secondary aim was to compare the accuracy of a newly derived prediction equation specific to the pediatric intensive care unit and the measured resting energy expenditure. DESIGN: A prospective, clinical, observational study. SETTING: A pediatric intensive care unit of a tertiary care medical center. PATIENTS: Forty-four children (29 males, 15 females) ages 2 wks to 17 yrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: During the course of their stay in the pediatric intensive care unit, 44 patients' measured resting energy expenditure was assessed using indirect calorimetry 94 times at up to three time points. The first measurement was at a mean time of 25 +/- 10 (+/-sd) hrs after admission, the second at 73 +/- 16 hrs, and the third immediately before discharge, which occurred at a mean of 193 +/- 93 hrs after admission. Measured energy expenditure varied only slightly (7% to 10%) from the first to second and the second to third measurements. Evidence for hypermetabolism was not apparent. Generally, the prediction equations performed well. Mean measured resting energy expenditure for all measurements was 821 +/- 653 kcals/24 hrs. The Schofield equation estimate was 798 +/- 595 kcals/24 hrs and the White equation estimate was 815 +/- 564 kcals/24 hrs (p = not significant). Nineteen (20%) measurements were >110% above the age-appropriate Schofield-predicted equation, and 30 measurements (32%) were <90% below that predicted by Schofield. Consequently, 45% of measured resting energy expenditure measurements were within 90% to 110% of that predicted by the Schofield equation. The White equation was inaccurate (not within 10% of measured resting energy expenditure) in 66 of 94 measurements (70%). The discrepancy was greatest (100%) in children with measured resting energy expenditure <450 kcal/24 hrs. CONCLUSION: The hypermetabolic response apparent in adults was not evident in these critically ill children. Currently available prediction equations cannot substitute for indirect calorimetry measurement of energy expenditure in guiding nutritional support in pediatric intensive care units. PMID- 17417118 TI - Identification of adrenal insufficiency in pediatric critical illness. AB - OBJECTIVE: To determine physicians' beliefs and practices regarding adrenal dysfunction in pediatric critical illness. DESIGN: Cross-sectional mail survey. SETTING: Canada. PARTICIPANTS: All members of the Canadian Pediatric Endocrine Group and all physicians identified as practicing pediatric intensive care medicine in any of 16 tertiary care teaching centers in Canada. INTERVENTIONS: Three pediatric intensive care physicians and three pediatric endocrinologists reviewed the questionnaire before administration to ensure clarity. We asked participants to report their views on the following: a) the frequency of adrenal insufficiency in pediatric critical illness; b) diagnosis/definition of adrenal insufficiency in pediatric critical illness; and c) the use of empirical glucocorticoids in fluid/vasopressor-resistant hypotension in pediatric critical illness. MEASUREMENTS AND MAIN RESULTS: Forty-six of 57 (80.7%) endocrinologists responded, with 43 participating (75.4%). Among intensivists, 59 of 70 (84.3%) responded with no refusals. Of intensivists, 81.4% believe that adrenal insufficiency occurs sometimes or often in critically ill pediatric intensive care unit patients, whereas 41.8% of endocrinologists believe adrenal insufficiency occurs never or rarely in these patients. Six definitions of adrenal insufficiency were proposed (varying cortisol level vs. peak/increment of cortisol in response to corticotropin), with no consensus on the definition of adrenal insufficiency from the endocrinologists or intensivists. Half (50.9%) of intensivists said they would sometimes or often empirically treat hypotensive pediatric patients with glucocorticoids, whereas 81.0% of endocrinologists would occasionally or never recommend glucocorticoids on this basis. CONCLUSIONS: There is no consensus among pediatric intensivists or endocrinologists as to how often adrenal insufficiency occurs in pediatric critical illness or how to diagnose this condition. Despite this lack of consensus, however, many pediatric intensivists would empirically treat hypotensive patients who they suspect may have adrenal insufficiency. Prospective studies are required to determine the definition, frequency, and appropriate treatment of adrenal insufficiency in critically ill pediatric patients. PMID- 17417119 TI - Mortality before and after initiation of a computerized physician order entry system in a critically ill pediatric population. AB - OBJECTIVE: A worrisome increase in mortality has been reported recently following the initiation of a computerized physician order entry (CPOE) system in a critically ill pediatric transport population. We tested the hypothesis that such a mortality increase did not occur after the initiation of CPOE in a pediatric population that was directly admitted to the neonatal and pediatric intensive care units at Montefiore Medical Center during two 6-month periods before CPOE and one 6-month period immediately after CPOE was initiated. Mortality in the pre and post-CPOE time periods was compared, and adjustment for potentially confounding covariates was performed. SETTING: The pediatric and neonatal intensive care units at Montefiore Medical Center. PATIENTS: All patients admitted from the emergency room or operating room or as transfers from other institutions directly to the pediatric and neonatal intensive care units at Montefiore Medical Center. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Overall, 29 (3.16%) of the 917 patients in the pre-CPOE period and nine (2.41%) of the 374 patients in the post-CPOE period died during their hospital stay (p = .466). The power to detect the hypothesized mortality increase was 81.7%. The variables that remained significant risk factors for mortality after adjustment were shock (odds ratio, 9.41; 95% confidence interval, 2.90-30.49), prematurity (odds ratio, 3.57; 95% confidence interval, 1.74-7.30), male gender (odds ratio, 3.31; 95% confidence interval, 1.47-7.69), or a hematologic/oncologic diagnosis (odds ratio, 3.14; 95% confidence interval, 1.44-6.86). Post-CPOE initiation status remained unassociated with mortality after adjusting for all covariates (odds ratio, 0.71; 95% confidence interval, 0.32-1.57). CONCLUSION: Mortality did not increase during CPOE initiation. PMID- 17417120 TI - Successful use of extracorporeal membrane oxygenation in severe necrotizing pneumonia caused by Staphylococcus aureus. AB - OBJECTIVE: To report the successful use of extracorporeal membrane oxygenation (ECMO) as rescue therapy for severe necrotizing pneumonia secondary to infection by the Staphylococcus aureus species. DESIGN: Case series. SETTING: Pediatric intensive care unit at a freestanding tertiary care children's hospital. PATIENTS: Two pediatric patients with severe S. aureus-induced necrotizing pneumonia requiring rescue with ECMO. Both patients survived with good neurologic outcomes. One patient required the use of activated factor VII for severe bleeding while on ECMO, with no thrombotic effect on the ECMO circuit. CONCLUSION: ECMO as rescue support should be considered in a timely fashion for refractory hypoxemic respiratory failure resulting from S. aureus pneumonia, including patients with necrotizing pneumonia. Use of ECMO support in such cases, coupled with aggressive measures aimed at minimizing bleeding, such as the use of activated factor VII, may result in excellent short- and long-term outcomes for such patients. PMID- 17417121 TI - High-frequency percussive ventilation in a pediatric patient with hydrocarbon aspiration. AB - INTRODUCTION: To describe ventilator management using a high-frequency percussive ventilator (HFPV), after other modes of mechanical ventilation failed. DESIGN: Case series. SETTING: Pediatric intensive care unit. PATIENTS: Previously healthy 11-month-old male with severe aspiration pneumonitis from mineral oil. INTERVENTIONS: The patient was initially placed on a conventional ventilator in a pressure-regulated volume-control mode but needed higher-than-normal pressures to maintain adequate ventilation. A decision was made to switch the patient to a pressure-control/pressure-support mode. At the end of the third day of pressure control/pressure-support mode, a decision was made to attempt airway pressure release ventilation. During a trial attempt, saturation levels deteriorated and a decision was made to place the patient on a high-frequency oscillator. The patient remained on this mode of ventilation for 6 days. On the sixth day, the chest radiograph showed a worsening of his pneumonia, and the patient started to deteriorate. A decision was made to try the HFPV in an attempt to mobilize secretions and any residual mineral oil. Immediately after initiating the HFPV and for 4 hrs thereafter, large amounts of secretions -- including a thick, oily substance -- were suctioned from the airways. Within 12-24 hrs, oxygenation improved dramatically and Fio2 was weaned. During the next 12 hrs, the patient was weaned off HFPV onto a conventional ventilator, and he was extubated 48 hrs after initiating HFPV. CONCLUSIONS: In this case, HFPV used as an alternative mode of ventilation successfully mobilized secretions that were otherwise unobtainable and that we believe led to the swift recovery of this child. HFPV should be given consideration as a mode of ventilation when mobilization of secretions is an issue. PMID- 17417122 TI - Tissue plasminogen activator for a left atrial thrombus after Senning repair. AB - OBJECTIVE: To avoid the surgical removal of an obstructive thrombus in a Senning baffle by the administration of recombinant tissue-type plasminogen activator. SETTING: A pediatric intensive care unit in a children's university hospital. PATIENTS: A 3-yr-old male was diagnosed with a large left atrial thrombus 2 wks after Senning repair for D-transposition of the great arteries. The child presented with massive chylous pleural, pericardial effusions, and cardiac tamponade, secondary to partial obstruction of the pulmonary venous channel. INTERVENTION: Thrombolysis with recombinant tissue-type plasminogen activator was instituted. RESULTS: We observed a resolution of the thrombus in <48 hrs. Minor local bleeding was the only noted side effect. No signs of systemic thromboembolization were detected. CONCLUSION: Early thrombolysis with recombinant tissue-type plasminogen activator could be considered a possible alternative to surgical thrombectomy in selected postoperative pediatric cases, although there may be a potential risk of serious bleeding. PMID- 17417123 TI - Subarachnoid-pleural fistula in an infant treated with mechanical positive pressure ventilation. AB - OBJECTIVE: To report a pediatric case of subarachnoid-pleural fistula, its diagnosis, and its treatment. DESIGN: Case report. SETTING: Pediatric intensive care unit. PATIENT: A 9-month-old boy, presenting with severe pleural effusion after posterior chest wall surgery. INTERVENTIONS: Subarachnoid-pleural fistula was confirmed by isolating beta2-transferrin in the pleural fluid and with magnetic resonance cisternography revealing the location. The patient had a healthy outcome, and the fistula dried without surgery, using positive-pressure ventilation and a chest drain. CONCLUSIONS: We used beta2-transferrin to confirm the diagnosis of clinically suspected subarachnoid-pleural fistula. High resolution computed tomographic and magnetic resonance cisternography are the best techniques to localize the fistula. The currently recommended treatment is surgery; we suggest that bilevel positive-pressure ventilation, especially with noninvasive techniques, could be a treatment alternative, reducing the flow of cerebral spinal fluid through the fistula and allowing spontaneous closure. PMID- 17417124 TI - Transhepatic Broviac catheter placement for long-term central venous access in critically ill children with complex congenital heart disease. AB - OBJECTIVE: Critically ill children with cardiac disease often require prolonged central venous access. Thrombosis of systemic veins or the need to preserve vessels for future cardiac procedures limits sites for placement of central venous catheters in these patients. This study evaluates the use of Broviac placement via the transhepatic approach for this patient population. DESIGN: A retrospective review. SETTING: A tertiary care center. PATIENTS: All children with complex congenital heart disease who underwent transhepatic Broviac placement between May 2000 and April 2004. INTERVENTIONS: Transhepatic Broviac placement. MEASUREMENTS AND MAIN RESULTS: Thirty-two children with a median age of 5 months (20 days-5.3 yrs) and a median weight of 4.2 kg (2.2-24.9 kg) underwent 40 transhepatic Broviac placements. There were three (8.8%) procedural related complications. One patient suffered an intra-abdominal bleed requiring an urgent laparotomy and removal of the Broviac, one patient required transfusion because of a mild self-contained intra-abdominal bleed, and one patient developed temporary complete heart block. There was one catheter infection. Thrombus was noted by echocardiography on the tip of two Broviacs; however, no intracardiac vegetations or embolic events occurred. There was no mortality related to the procedure. Broviacs remained in place for a median of 36 days (1 day-6 months). Five Broviacs were dislodged inadvertently (two during cardiac massage and three resulting from patient manipulation). The remaining Broviacs were electively removed safely without coil embolization. At a median follow-up of 3.5 months (10 days-3 yrs), there have been no long-term complications related to the Broviacs. CONCLUSIONS: Transhepatic Broviac catheters can be used safely in critically ill children with cardiac disease and remain indwelling for adequate periods of time. This modality of prolonged vascular access should be considered for children whose veins are occluded or need to be preserved for future procedures. PMID- 17417125 TI - Pediatric staff perspectives on organ donation after cardiac death in children. AB - OBJECTIVES: The aims of this project were to describe whether pediatric clinical staff members believe that a donation after cardiac death (DCD) program could be consistent with the mission and core values of a children's hospital and to identify what staff consider essential to the acceptability of such a program. DESIGN: Qualitative study. SETTING: Children's hospital. SUBJECTS: Pediatric clinical staff. INTERVENTIONS: Data were gathered from pediatric clinical staff during eight focus groups conducted in a children's hospital in March and April 2005. MEASUREMENTS AND MAIN RESULTS: Eighty-eight staff members participated. Six major themes emerged from qualitative analysis of the data: a) identifying children who could be candidates for DCD; b) considering the best interests of the dying child; c) approaching parents about DCD; d) preparing parents for their child's DCD; e) doing DCD well; and f) maintaining program integrity. Themes were used to construct a conceptual framework describing a model pediatric DCD program. Pediatric staff voiced numerous concerns. However, they identified "making it happen for families" who voice a desire to participate in organ donation as the primary reason for program adoption. CONCLUSIONS: This study provides a framework for understanding pediatric staff perspectives on DCD programs in children. Results suggest several possible elements that may be helpful in framing interdisciplinary dialogue and informing institutional practices in the design of a pediatric DCD program. PMID- 17417126 TI - Preventable harm occurring to critically ill children. AB - OBJECTIVE: To develop a trigger tool for identifying adverse events occurring in critically ill pediatric patients; to identify and characterize adverse events and preventable adverse events experienced by critically ill pediatric patients; and to characterize the patients who experience preventable adverse events. DESIGN: Retrospective chart review using a trigger tool. SETTING: Pediatric intensive care unit of a tertiary, university-affiliated pediatric hospital. PATIENTS: A systematic sample of 259 pediatric intensive care unit patients from a 1-yr period. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured frequency of occurrence (0.19 preventable adverse events per patient-day), severity of harm (78% minor, 19% moderate, 3% serious, no deaths), and type of event (sedation, 22%; skin, 16%; medical device complication, 14%; pulmonary, 13%; and cardiovascular, 11%). Patients who experienced preventable adverse events were younger, had longer lengths of stay, and had higher illness burdens. Preventable adverse events occurred more frequently among surgical patients than medical patients. CONCLUSIONS: Preventable adverse events occurred fairly frequently in the pediatric intensive care unit, but serious harm was rare. Conditions that increased the likelihood of a preventable adverse event were a) need for sedation or pain control; b) relative immobility; and c) need for vascular devices, feeding tubes, or ventilators. Adverse event prevention strategies that focus on improving patient monitoring under increased-risk conditions and improving early detection and treatment of potential harm will likely be more effective than strategies aimed at general error prevention. PMID- 17417127 TI - Increased calcium supplementation is associated with morbidity and mortality in the infant postoperative cardiac patient. AB - OBJECTIVE: The purpose of this study was to assess the association of calcium replacement therapy with morbidity and mortality in infants after cardiac surgery involving cardiopulmonary bypass. DESIGN: Retrospective chart review. SETTING: The cardiac intensive care unit at a tertiary care children's hospital. PATIENTS: Infants undergoing cardiac surgery involving cardiopulmonary bypass between October 2002 and August 2004. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Total calcium replacement (mg/kg calcium chloride given) for the first 72 postoperative hours was measured. Morbidity and mortality data were collected. The total volume of blood products given during the first 72 hrs was recorded. Infants with confirmed chromosomal deletions at the 22q11 locus were noted. Correlation and logistic regression analyses were used to generate odds ratios and 95% confidence intervals, with p < .05 being significant. One hundred seventy one infants met inclusion criteria. Age was 4 +/- 3 months and weight was 4.9 +/- 1.7 kg at surgery. Six infants had deletions of chromosome 22q11. Infants who weighed less required more calcium replacement (r = -.28, p < .001). Greater calcium replacement correlated with a longer intensive care unit length of stay (r = .27, p < .001) and a longer total hospital length of stay (r = .23, p = .002). Greater calcium replacement was significantly associated with morbidity (liver dysfunction [odds ratio, 3.9; confidence interval, 2.1-7.3; p < .001], central nervous system complication [odds ratio, 1.8; confidence interval, 1.1 3.0; p = .02], infection [odds ratio, 1.5; confidence interval, 1.0-2.2; p < .04], extracorporeal membrane oxygenation [odds ratio, 5.0; confidence interval, 2.3-10.6; p < .001]) and mortality (odds ratio, 5.8; confidence interval, 5.8 5.9; p < .001). Greater calcium replacement was not associated with renal insufficiency (odds ratio, 1.5; confidence interval, 0.9-2.3; p = .07). Infants with >1 sd above the mean of total calcium replacement received on average fewer blood products than the total study population. CONCLUSIONS: Greater calcium replacement is associated with increasing morbidity and mortality. Further investigation of the etiology and therapy of hypocalcemia in this population is warranted. PMID- 17417128 TI - Institutional policies on determination of medically inappropriate interventions: use in five pediatric patients. AB - OBJECTIVE: To describe recent experience using the Texas Advance Directives Act to facilitate care of terminally ill children managed in the two tertiary pediatric hospitals of the Texas Medical Center, Houston, TX. DESIGN: Retrospective chart review. SETTING: Two multidisciplinary pediatric intensive care units in Houston, TX. PATIENTS: Five terminally ill children whose parents were unable to acquiesce to comfort or palliative care. INTERVENTIONS: Implementation of the Texas Advanced Directives Act of 1999. RESULTS: Suspension of interventions thought to be medically inappropriate by the physicians of record in four of the five cases, with transfer of care in one instance. CONCLUSIONS: Use of institutional policies in accordance with the Texas Advance Directives Act may assist in the care of terminally ill children and their families. PMID- 17417129 TI - Assessment of parental presence during bedside pediatric intensive care unit rounds: effect on duration, teaching, and privacy. AB - OBJECTIVE: There is a paucity of literature evaluating the effects of family member presence during bedside medical rounds in the pediatric intensive care unit. We hypothesized that, when compared with rounds without family members, parental presence during morning medical rounds would increase time spent on rounds, decrease medical team teaching/education, increase staff dissatisfaction, create more stress in family members, and violate patient privacy in our open unit. DESIGN: Prospective, blinded, observational study. SETTING: Academic pediatric intensive care unit with 12 beds. PARTICIPANTS: A total of 105 admissions were studied, 81 family members completed a survey, and 187 medical team staff surveys were completed. INTERVENTIONS: Investigators documented parental presence and time allocated for presentation, teaching, and answering questions. Surveys related to perception of goals, teaching, and privacy of rounds were distributed to participants. MEASUREMENTS: Time spent on rounds, time spent teaching on rounds, and medical staff and family perception of the effects of parental presence on rounds. RESULTS: There was no significant difference between time spent on rounds in the presence or absence of family members (p = NS). There is no significant difference between the time spent teaching by the attending physician in the presence or absence of family members (p = NS). Overall, parents reported that the medical team spent an appropriate amount of time discussing their child and were not upset by this discussion. Parents did not perceive that their own or their child's privacy was violated during rounds. The majority of medical team members reported that the presence of family on rounds was beneficial. CONCLUSIONS: Parental presence on rounds does not seem to interfere with the educational and communication process. Parents report satisfaction with participation in rounds, and privacy violations do not seem to be a concern from their perspective. PMID- 17417131 TI - Abnormalities of coagulation related to the use of inhaled nitric oxide before extracorporeal membrane oxygenation. AB - OBJECTIVE: Evaluation of the influence of previous inhaled nitric oxide (iNO) treatment on the occurrence of clotting complications and disseminated intravascular coagulation during extracorporeal membrane oxygenation (ECMO). DESIGN: Retrospective study in newborns treated with venoarterial ECMO during a 5 yr period. SETTING: Neonatal intensive care unit of a university medical center. PATIENTS: A total of 59 newborns with severe respiratory insufficiency treated with venoarterial ECMO. INTERVENTIONS: Patients received iNO before ECMO (iNO group) or not (control group). MEASUREMENTS AND MAIN RESULTS: There were no differences between the groups for patient characteristics and medication use before ECMO, except for norepinephrine. After correction for diagnosis and duration of ECMO, significantly more clotting complications and disseminated intravascular coagulation as individual variables were seen in the iNO group. For the combination of clotting complications and disseminated intravascular coagulation, there was a significantly higher prevalence in the iNO group. CONCLUSIONS: In our population, we found a remarkable relationship between clotting complications or disseminated intravascular coagulation and iNO use before ECMO treatment, which needs further prospective research before conclusions can be drawn. PMID- 17417132 TI - Cervical infection with Chlamydia trachomatis and Neisseria gonorrhoeae in women from ten areas in four continents. A cross-sectional study. AB - OBJECTIVES: Better information on the prevalence of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infection is needed in many world areas. STUDY DESIGN: Cross-sectional study of population-based samples of nonpregnant women aged 15 to 44 years in Nigeria, Colombia, Argentina, Vietnam (2 areas), China, Thailand (2 areas), Korea, and Spain. 5,328 consenting women aged 15 to 44 years participated. Exfoliated cervical cells were collected and testing for CT and NG and human papillomavirus (HPV) was done using PCR-based assays. RESULTS: Age standardized CT prevalence ranged between 0.2% (95% confidence interval, CI: 0.0 0.7%) in Spain and 5.6% (95% CI: 3.4-7.8%) in Nigeria. NG ranged between 0% (with broad CIs) in several areas and 2.6% (95% CI: 1.0-4.2%) in Nigeria. Prevalence of CT in all areas combined was greater in women aged 15 to 24 (4.5; 95% CI: 3.4 5.8%) than 25 to 44 (2.6; 95% CI: 2.1-3.1%), whereas NG prevalence was similar in the 2 age groups (0.3%). The only significant risk factors were NG infection (for CT), CT infection (for NG) and infection with high-risk HPV types (for both). CONCLUSIONS: The prevalence of CT and, most notably, NG was relatively low in a variety of countries. Our findings, however, do not apply to subsets of high-risk women who are likely to be underrepresented in our population-based samples. PMID- 17417133 TI - Self-reported mechanical problems during condom use and semen exposure. Comparison of two randomized trials in the United States of America and Brazil. AB - OBJECTIVES/GOAL: To compare self-reported condom use problems and objectively determined semen exposure in 2 populations. STUDY DESIGN: Two randomized crossover trials in the United States and Brazil compared the failure rates of the female condom (FC) and male condom (MC). Participants used both condom types, completed condom-specific questionnaires to report problems, and collected precoital and postcoital samples of vaginal fluid. Prostate-specific antigen (PSA) was detected by immunoassay. RESULTS: Problems with condom use were reported less frequently in the Brazilian study (rate difference: FC = 24%, P <0.0001, MC = 5%, P = 0.003). By contrast, the PSA detection rates were similar for both the FC and the MC (rate difference: FC = 2%, MC = 1%, not significant). These results suggest that the PSA detection rate was similar in the 2 study groups and that self-reported problems may be a less reliable measure of condom failure. CONCLUSIONS: Use of biomarkers of condom failure like PSA may help to strengthen the validity of studies promoting behavior change for the prevention of sexually transmitted diseases. PMID- 17417134 TI - [Protein transduction, from technology to physiology]. AB - In the early 90s, we found that the DNA-binding domain (homeodomain) of Antennapedia, a homeoprotein transcription factor, was internalized by live cells gaining access to their cytoplasm and nuclei. It was soon revealed that internalization is due to the third helix of the homeodomain, composed of sixteen amino acids. This short peptide baptized Penetratin is the first of a large series of transduction peptides widely used for the internalization of all sorts of cargoes in vitro and in vivo. Although transduction peptides are being developed with the latter practical goal, the most intriguing outcome of our initial observation is that full-length homeoproteins are transferred between cells and have non-cell autonomous transcriptional and translational activities. This new signaling mechanism requires that homeoproteins be internalized and secreted. Secretion is Golgi independent and requires a small sequence also present in the homeodomain but distinct from the Penetratin sequence. The consequences of this novel signaling mechanism are briefly discussed. PMID- 17417135 TI - [Tracking Trojan peptides in cells]. AB - Trojan peptides or cell-penetrating peptides (CPP) are natural or designed peptides identified as cellular membrane-crossing molecules, in particular through their potency to vehiculate various kinds of compounds to the cytoplasm and nucleus of living cells. The indirect methods used so far to detect these peptides in cells led to controversial hypotheses on the mechanism of their cell entry. Therefore, we have developed a MALDI-TOF mass spectrometry-based quantification method to track these peptides inside cells. This new method is presented in this review. PMID- 17417136 TI - [Transcription factor movement in plants]. AB - In the last few years, the intercellular trafficking of regulatory proteins has emerged as a novel mechanism of cell-to-cell communication in plant development. Here I present a review of the documented cases of transcription factors movement in plants and examine the common themes underlying these different examples. PMID- 17417137 TI - [Transduction peptides: structural-functional analyses in model membranes]. AB - Peptide-membrane interaction is the first step required for peptide cell internalization. In this paper we studied the interactions of substance P, Penetratin and an amphiphilic 16mer (RL16) peptide in two different model membranes, giant unilamellar vesicles and large unilamellar vesicles. Penetratin was able to induce the formation of tubes inside the giant vesicles without changes in membrane permeability. On the contrary, RL16 induced the disruption of giant vesicles and the permeabilization of large vesicles. Substance P showed none of these effects. PMID- 17417138 TI - [Ex vivo expansion of human hematopoietic stem cells by passive transduction of the HOXB4 homeoprotein]. AB - Expansion of human hematopoietic stem cells (HSCs) is a challenge for cellular therapy. It currently relies on either the use of recombinant cytokines or transfer of transcription factor genes. Among these, the HOXB4 homeoprotein is of particular interest since it promotes the expansion of mouse HSCs without inducing leukemia. To prevent potential deleterious side effects associated with stable HOXB4 gene transfer into the cells, we took advantage of the ability of homeoproteins to passively pass through cell membranes. We have shown that, when co-cultured with stromal cells engineered to secrete HOXB4, human stem cells and immature progenitors clearly were expanded. This expansion was associated with enhanced stem cell repopulating capacity in vivo and maintenance of pluripotentiality. The role that HOXB4 plays on stem cell expansion has also been tested on human lymphoid progenitors. We found that our model of protein transfer was also able to induce the expansion of the immature lympho-myeloid and pro-T/NK progenitors as well as of more mature NK progenitors. We then looked for synergistic activities between HOXB4 and other homeoproteins such as HOXC4. We found that HOXC4 was able to promote the expansion of human hematopoietic cells in vitro roughly as HOXB4 did and that the presence of both HOXB4 and HOXC4 molecules induced even higher expansion levels of these cells. Our method provides a basis for developing cell therapy strategies using expanded HSCs that are not genetically modified. PMID- 17417139 TI - [Paul Bert, scientist and politician]. AB - Paul Bert worked with Claude Bernard, one of the leading physiologists of the 19th century. In his laboratory at the College de France in Paris, Paul Bert carried out fascinating experiments in particular on respiratory processes, leading him to publish "La pression barometrique" in 1878. In this book are recalled his discovery of oxygen pressure decrease with altitude, divers diseases, the improved safety protocols in hyperbaric conditions, and the first development of gas anaesthetics for surgery. He was the third President of the Societe de Biologie. Paul Bert was also a politician with strong convictions. Minister of Education under Gambetta's short term government, he initiated the fight for social equality and secular education and became one of the most prestigious figures of the developing socialist party. He received many distinctions and was given a state funeral. PMID- 17417140 TI - [The exposure of man to altitude when flying: from Paul Bert to today]. AB - This paper is organised around Paul Bert's inputs to modern aviation physiology. Paul Bert gave a masterly historical work, which established the notion of oxygen partial pressure as the right physiological parameter for the body. Human protection against aviation altitude takes this notion into account, by modifying either the pressure surrounding the body or the oxygen fraction in inhaled gases. We also consider the case of protection against accidental loss of aircraft pressurization. PMID- 17417141 TI - [Diving: barometric pressure and neurochemical mechanisms]. AB - The studies of Paul Bert, presented in his book "La Pression Barometrique" in 1878, were at the origin of the modern hyperbaric physiology. Indeed his research demonstrated the effects of oxygen at high pressure, that compression effects must be dissociated from decompression effects, and that neurological troubles and death of divers during or after decompression were due to the fast rate of decompression. However, it is only in 1935 that the work of Behnke et al. attributed the complaints reported at 3 bars and above in compressed air or nitrogen-oxygen mixture to the increase in partial pressure of nitrogen which induces nitrogen narcosis. Little is known about the origins and mechanisms of this narcosis. The traditional view was that anaesthesia or narcosis occurred when the volume of a hydrophobic membrane site was caused to expand beyond a critical amount by the absorption of molecules of a narcotic gas. The observation of the pressure reversal effect during general anaesthesia has long supported this lipid theory. However, recently, protein theories have met with increasing recognition since results with gaseous anaesthetics have been interpreted as evidence for a direct gas-protein interaction. The question is to know whether inert gases, that disrupt dopamine and GABA neurotransmissions and probably glutamatergic neurotransmission, act by binding to neurotransmitter protein receptors. PMID- 17417142 TI - [From 1878 to 2006 - working in hyperbaric conditions during tunnelling]. AB - To review the impact of Paul Bert's researches on hyperbaric work in tunnelling, the status of the industry in 1878 is described. Mostly based on the application of Triger's machine it was used to mine coal below the water table or to dig foundations for bridges in rivers or close to rivers. The results and conclusions obtained by Paul Bert which are applicable in that particular field are listed. The major steps of research or remarkable achievements in construction between 1878 and 2006 are presented as well as the evolution of decompression tables. Improvement in safety and conditions of caisson workers has been continuous until the technical revolution resulting from the introduction and the development of tunnelling boring machines (TBM) in the late 80's. TBM technology has resulted in major changes in tunnel construction. Hyperbaric interventions have also changed completely since human operators no longer work in pressurized conditions. Only occasional inspections and repairs are carried out under pressure. Present performance in hyperbaric conditions are reported, and high pressures reached in the 2000's using saturation technology are described. The future of hyperbaric works is also discussed whether for very high pressure, or complete replacement of caisson workers in TBMs. These descriptions show that Paul Bert provides us with very clear directions to improve safety in hyperbaric conditions and that none of his recommendations were mistaken, most being still relevant. PMID- 17417143 TI - [How to live together with our differences]. AB - Analysing Paul Bert's contributions to the creation of a public, secular and compulsory school and also the meaning of the French 1905 law which separates Churches from the State, this contribution suggests to enlarge the concept of secularity from a religious to a non-religious acception, which opens on a new conception of the European Union. PMID- 17417144 TI - [A case report of a recurrence of Mooren's ulcer after cataract surgery]. AB - We report the case of Mooren's ulcer recurrence after uncomplicated cataract surgery in a 61-year-old woman. This cataract developed because of repetitive inflammation of the anterior chamber and corticotherapy. Local and general corticotherapy with cyclosporin 2% drops was started in association with an anterior lamellar graft and a conjunctival recession due to a preperforation condition. Secondarily cyclophosphamide was necessary to control recurrence with a good anatomic result and an increase in visual acuity. The case updates physiopathologic and diagnostic data on this rare limbic autoimmune ulcerative disease. The diagnosis was made by histology and the dosage of specific autoantibodies against cornea. The prevention of recurrence after surgery requires a long clinical quiescent period, minimally invasive surgery long after inflammation has subsided, and a gradual tapering of corticotherapy over several weeks. PMID- 17417145 TI - [Prepapillary arterial loop and retinal arterial branch occlusion]. AB - Preretinal arterial loops are congenital vascular anomalies that originate from a main branch of the central retinal artery on the optic disc. These arterial loops are usually unilateral and asymptomatic, but they can be associated with retinal artery branch occlusion. We report one case of inferior temporal retinal artery occlusion in a patient with preretinal arterial loops. Two different mechanisms are thought to be the cause of occlusion: twisting of the loop or thrombosis. PMID- 17417146 TI - [Utility of the Visante OCT in the follow-up of glaucoma surgery]. AB - PURPOSE: To evaluate the utility of the Visante OCT in the follow-up of filtering surgery. MATERIAL: and methods: In this study, we evaluated 38 filtering blebs in 31 patients following trabeculectomy (4) and nonpenetrating deep sclerectomy (NPDS). In 11 cases, mitomycin C was used during surgery. In patients who underwent NPDS, a collagen implant was placed within the scleral site in eight eyes and seven eyes had goniopuncture during follow-up. All patients had complete ophthalmologic examination with morphological evaluation of the bleb and intraocular pressure measurement as well as Visante OCT evaluation. RESULTS: The trabeculectomy site as well as the trabeculodescemetic membrane (in NPDS), the scleral flap, the conjunctival flap, the iris, and the relationship between these structures were analyzed. Functioning blebs had a hyporeflective and irregular conjunctival tissue associated with a route for aqueous humor under the scleral flap from the anterior chamber toward the subconjunctival space. This filtration route was also observed within nonfunctioning encapsulated blebs. Flat and encapsulated nonfunctioning blebs had dense and hyperreflective conjunctival tissue. In the particular case of nonfunctioning flat blebs, there was no route for aqueous humor filtration under the scleral flap. CONCLUSION: The Visante OCT is a simple and noninvasive imaging technique with good resolution, allowing the analysis of morphologic changes occurring in eyes after filtering surgery. PMID- 17417147 TI - [Quality of life in patients with wet age-related macular degeneration treated with photodynamic therapy]. AB - PURPOSE: To analyze the benefit of photodynamic therapy in terms of quality of life in patients with wet age-related macular degeneration. METHODS: A retrospective study was conducted on 33 patients with subretinal neovascularization receiving visual rehabilitation in a low vision clinic (ARAMAV, Nimes) in southern France. Twenty had been treated with photodynamic therapy (PDT group) and 13 had not (non-PDT group). In the non-PDT group, the patients had been treated with thermal photocoagulation, transpupillary thermotherapy, or external radiotherapy, or had not received any treatment. Visual acuity, reading speed, reading endurance, and quality of life were compared. Two quality-of-life scales were employed: the VF-14 and NEI-VFQ-25. RESULTS: A significant difference between the two groups in terms of quality of life was observed. The VF-14 score (p<0.01) and two parameters (near activities and distant activities) of the NEI VFQ-25 were significantly higher (both p<0.01) in the PDT group. Visual acuity, reading speed, and reading endurance were similar in both groups. CONCLUSION: Although no difference in terms of functional parameters was observed, photodynamic therapy could preserve the central retina and thus enable a better use of the residual visual function, which could explain the better quality of life perceived by the patients in the PDT group. PMID- 17417148 TI - [A new approach for better comprehension of diseases of the ocular surface]. AB - The mechanistic view of dry eye disease aims at completing the classic etiological approach that classifies the disease as parallel ocular surface disorders leading to lacrimal film impairment and dry eye. This approach proposes two levels of ocular surface impairment (with standard etiologies, previously validated in the NEI/Industry workshop), which may not be independent diseases but rather risk factors and/or ways to enter a self-stimulated biological process involving the ocular surface. All external disorders proposed in this model, although unlikely to be fully exhaustive, are classical mechanisms considered to be causes of tear film impairment and ocular surface damage, by tear instability and evaporation, tear hyposecretion, or both. These mechanisms, sometimes alone- when severe or becoming chronic or repeatedly present on the ocular surface and when two or more are present--may cause the patient to enter the self-stimulated loop. Tear film instability/imbalance can be considered as the key point of dry eye disease. It will cause local or diffuse hyperosmolarity of the tear film and therefore of superficial epithelial cells of the cornea and/or conjunctiva, stimulating epithelial cells and resident inflammatory cells. Cell damage in the cornea and conjunctiva, by means of apoptosis and direct mechanical and/or osmotic stress, will stimulate the reflex neurosensory arc, in turn stimulating lacrimal gland and neurogenic inflammation, with inflammatory cytokine release, MMP activation, and inflammatory involvement of the conjunctival epithelium. Goblet cell loss is thus directly related to chronic inflammation and surface cell apoptosis subsequent to cell hyperosmolarity and chronic damage, resulting in further tear film instability/imbalance. On the other hand, bacterial changes and an imbalance resulting from specific diseases or from tear film abnormalities may trigger release of endotoxins, lipopolysaccharides, and/or lipase activation, causing eyelid inflammation, meibomian gland dysfunction, and lipidic changes, directly influencing tear film stability and favoring tear evaporation. The lipidic hypothesis therefore participates in the vicious circle as a parallel, independent, or complementary loop. This mechanistic approach proposes a synthetic combination of mechanisms previously validated independently, with two levels of ocular surface impairment, a first level including many possible acute or chronic causes that favor or trigger the imbalance and can be reversible if correctly and specifically managed when possible, and the further involvement of a series of biological cascades centered by tear film imbalance and inflammatory stimulation, finally acting as an independent vicious circle, however the patient entered the loop. Clinically, this approach may explain examples of dry eye syndrome occurring after ocular surgery, contact lens wear, chronic allergy or systemic or topical drugs, and the long-lasting effect even though all causal factors have been removed or have disappeared. This model should be considered as a basis for further reflection on biological mechanisms that could be even more complex but individually constitute potential leads for targeting therapeutic strategies to allow patients to leave the loop even though the triggering factors are still present or can only be attenuated, such as in Sjogren syndrome or ocular rosacea. It also should be considered a complement to more classic etiological and severity classifications aimed at understanding and classifying the large number of diseases that may cause dry eye disease and better assessing the major impairment it causes on the patient's quality of life. PMID- 17417149 TI - [Discoid lupus erythematosus with eyelid involvement. A series of nine patients]. AB - INTRODUCTION: Discoid lupus erythematosus (DLE) is a chronic autoimmune skin disease that usually affects the sun-exposed skin. Palpebral involvement occurs uncommonly. MATERIAL AND METHOD: The goal of our study was to present the clinical and therapeutic features of a series of nine patients with discoid lupus erythematosus with eyelid involvement. RESULTS: Lesions of discoid lupus were more frequent in the lower eyelids (seven cases). The palpebral location was the only manifestation of the disease in a 34-year-old woman. In the other cases, cutaneous lesions of typical discoid lupus were noted. Seven patients responded to therapy with antimalarial drugs associated with local corticosteroids and photoprotection. DISCUSSION: Eyelid lesions of discoid lupus erythematosus are rare. Involvement of the lower eyelids is more common. It is important to diagnose discoid lupus of the eyelids because misdiagnosis (isolated form) can delay treatment and cause deformities. The treatment is systemic antimalarial drugs, which have an excellent clinical response. PMID- 17417150 TI - [Late endophthalmitis complicating glaucoma filtering surgery without adjunctive antifibrotic agents]. AB - PURPOSE: To identify the incidence, infecting organisms, and clinical results of late endophthalmitis after trabeculectomy without antifibrotic agents. MATERIAL AND METHODS: The current retrospective study investigated 613 trabeculectomies without adjunctive antifibrotic agents performed between January 1994 and December 2003 in the Clinic of Ophthalmology of Beni-Messous University Hospital (Algiers, Algeria). RESULTS: Eight eyes presented with late endophthalmitis after an uneventful postoperative period, for an incidence of 1.3%. The endophthalmitis occurred between 1 year and 7 years after the filtering surgery. Conjunctival swabs were taken in all cases. Vitreous swabs were taken in only six cases. There were five positive cases. The organisms identified were Staphylococcus aureus (one case), alpha hemolytic streptococci (two cases), Haemophilus sp. (one case), and Pseudomonas aeruginosa (pyocyanic) (one case). The anatomic and visual results were poor after treatment of endophthalmitis. One case deteriorated, ending in enucleation. CONCLUSION: Late endophthalmitis may complicate glaucoma filtering surgery, several months or years later, after a postoperative period without incident. The surgical indications of glaucoma must take into account the complications related to this surgery. PMID- 17417151 TI - [Management of hyperopia in children]. AB - PURPOSE: To study the prevalence of hyperopia in school-aged children and to analyze the factors that increase the risk of squint or amblyopia in a retrospective study. METHODS: Three hundred eyes of 150 children with hyperopia who did not have anisometropia > or =1.5 D were selected. Complete ophthalmological examination was performed for all children. Hyperopia was defined when spherical equivalent was +0.5 D or greater. Amblyopia was screened and treated by patching therapy and then penalisation. Complete spectacle correction was achieved in children with high hyperopia (+3.5 D or greater) or in presence of squint or amblyopia. A statistical analysis compared the results using the Mann-Whitney test and the chi square test. RESULTS: The mean age was 9.5+/-2.7 years. Girls were statistically more represented than boys. The mean sphere measured overall was +2 D (+/-1.65). Severe hyperopia was detected in 19% of the children; it was latent in 35% of children. Strabismus was detected in 7% and was accommodative in 25%. Esotropia was the most prevalent deviation (72.8%). The prevalence of amblyopia was 12%. The mean sphere measured in amblyopic children was 5.66 D (+/-1.64 D). Initial depth of amblyopia was mild to moderate and 98% of the children achieved iso-acuity after patching therapy. The correlation between severe hyperopia, amblyopia, and squint was statistically significant. Indeed, the risk ratios of squint and amblyopia, 5.2 and 3.70, respectively, were significantly high in children with severe hypermetropia. Complete spectacle correction improved final visual acuity and reduced the angle deviation in accommodative esotropia. CONCLUSION: Children with hyperopia of +3.5 D or greater have an increased risk of amblyopia and squint that threatens their visual function. Hyperopic correction should be prescribed even if no strabismus or amblyopia is detected in order to prevent this risk. Screening programs should also be promoted to detect these children at an early age. PMID- 17417152 TI - [Clinical evaluation of the Pascal dynamic contour tonometer]. AB - PURPOSE: The Pascal dynamic contour tonometer (DCT) was designed to measure IOP independently of corneal properties. This study aimed at 1) assessing the intra- and interindividual variability of DCT IOP measurements, the differences between DCT and applanation tonometry IOP measurements (APL), and their correlations with central corneal thickness (CCT); 2) analyzing the variability of the ocular pulse amplitude (OPA) and its correlations with age, blood pressure (BP), cardiac beat pulse (CP), diagnosis of glaucoma, IOP, and severity of glaucomatous visual field (VF) defects. METHODS: Twenty-five normal subjects (25 eyes), 14 patients with ocular hypertension (27 eyes), and 54 glaucomatous patients (104 eyes) were included in this prospective study. In the first 12 normal subjects, three consecutive IOP measurements were taken by three different observers using DCT, directly followed by three measurements with APL by the same observer. In the following 13 subjects, the reverse sequence was followed. In the other group, the IOP measurements (three DCT and three APLs) were taken by the same observer. Only DCT measurements with quality levels 1-3 were considered for analysis. RESULTS: In the normal group, DCT IOP measurement variability varied between 4.4%-7.3% (intraobserver variation coefficient) and 8% (interobserver variation coefficient). DCT IOP measurement was not influenced by the sequence of measurements or the observer. DCT overestimated IOP by a mean of 2.2 mmHg compared with APL (p<0.001). The 95% limits of agreement for each subject tested with both tonometers ranged from -0.5 mmHg to +6.3 mmHg. IOP APL and DCT measurements were strongly correlated. Both DCT and APL were not correlated with CCT. OPA ranged from 1.2 mmHg to 6.6 mmHg (mean, 3.1+/-1.2 mmHg) and was comparable between the three observers. Intraobserver OPA variability ranged from 7.6% to 9.5%. The interobserver OPA variability coefficient was 8.8%. OPA was only correlated with systolic BP (p<0.05). In glaucomatous patients, the correlation between DCT and APL IOP measurements was highly significant (r=0.860, p<0.001). DCT overestimated IOP by a mean 2 mmHg compared with APL (p<0.001). IOP differences between both tonometers were not influenced by the sequence of measurements. Unlike APL, DCT was not or only slightly influenced by CCT (p=0.07 for DCT; p=0.001 for APL). The mean difference between IOP DCT and APL was larger in thin corneas (<520 microm): 2.8+/-3.1 mmHg versus 0.8+/-2.3 mmHg in thick corneas (580 microm) (p=0.001). OPA was not correlated with age. It was positively correlated with IOP (p<0.001), systolic BP (p=0.047), and MD (mean deviation) (p=0.018). It was negatively correlated with diastolic BP (p=0.003), cardiac frequency (p<0.001), severity of glaucomatous VF defects (p=0.002), and PSD (pattern standard deviation) (p=0.008). It was significantly higher in the OHT subgroup and significantly lower in the NTG subgroup (p<0.05). In both groups, the IOP difference between DCT and APL was not correlated with age (p>0.05). CONCLUSIONS: IOP measurements with the Pascal(R) DCT and APL correlated well and were reproducible. DCT IOP measurement variability was slightly higher than APL with relatively wide 95% limits of agreement. Considering the entire study population, DCT overestimated IOP by a mean 2.0 mmHg compared with APL. DCT was independent of CCT, especially in thin corneas. The DCT does not appear to be clinically advantageous over the Goldmann tonometer in the IOP measurement in thick corneas. Therefore an IOP follow-up by APL tonometry and pachymetry appeared to be mandatory for the interpretation of the true IOP. Interindividual OPA variations were high, as was measurement variability. OPA was correlated with BP, cardiac frequency, IOP, diagnosis of glaucoma, and severity of glaucomatous VF defects. These must be considered in the clinical interpretation of this parameter. PMID- 17417153 TI - [Improved visualization of fundus with green-light ophthalmoscopy]. AB - Monochromatic light accentuates details of different retinal layers because of its variable absorption and reflectance by structures both within and above these layers. Red-free light is little used, although it is an elementary method. Green light ophthalmoscopy, with its short wavelength, enhances some fundus and vitreous structures and may make the examination of pathologic conditions (premacular pathology, vascular abnormalities, etc.) easier. Furthermore, it is often more comfortable for the patient. PMID- 17417154 TI - [Optical coherence tomography follow-up of a case of solar maculopathy]. AB - Solar maculopathy is caused by sun-gazing without protection and in most cases appears to be reversible. Retinal damage may be caused by photochemical changes combined with a rise in temperature at the time of sun observation. We report a case of bilateral solar retinopathy following direct sun gazing after observation of the partial eclipse on 3 October 2005. A 25-year-old Caucasian man presented 24 h after watching the eclipse with symptoms of blurred vision and a bright positive central scotoma. Total direct viewing time was approximately 5 min. While observing the eclipse, the patient kept both eyes open. On presentation, his visual acuity was 20/25 in his right eye and 20/32 in his left eye. Ophthalmoscopic examination revealed a bilateral yellowish-white spot in the center of the foveal region. Static visual field examination did not disclose any absolute scotoma but a decrease in the foveal threshold was noted in both eyes. Optical coherence tomography (OCT) examination revealed a bilateral increased reflectivity of the inner foveal retina that was greater in the left eye. This finding was associated with a hyporeflective area of the underlying retinal pigment epithelium: choriocapillaris complex and an increase in retinal thickness. Eight days later, visual acuity was 20/20 in each eye. Funduscopy showed the yellow lesion had vanished. Four months after exposure, funduscopy was normal and OCT scan showed normal reflectivity of all retinal layers in the fovea. However, static visual field examination disclosed a decreased foveal threshold on the left eye. We compare the OCT findings in this patient with the data reported in the literature. Indeed, various OCT findings have been reported depending on the intensity and frequency of sun exposure. This observation demonstrates that OCT appears to be potentially useful in the evolutive follow-up of solar maculopathy, and emphasizes the importance of eye safety during solar eclipse observation. PMID- 17417155 TI - [The cellular players in allergy]. AB - Ocular allergy, with a significant increase of prevalence, is one of the most usual disease of the ocular surface. But, in a same entity, it means different kinds of clinical diseases with acute hayfever conjunctivitis, usually easy to diagnose, but also chronic perennial forms without any functional or clinical specificity among others ocular surface diseases. These clinical aspects are underlain by complex cellular mecanisms which are essentially located inside the conjunctiva. Initially, allergen cross the lacrimal tear film, the first natural barrier of the eye, then in the conjunctival epithelium, meet the antigen presenting cells, the Langerhans cells, lymphocytes and all the others components of the inflammatory and allergic reaction, including mast cells and eosinophils. The cornea could also be involved in allergic diseases, mostly in severe ones such as vernal or atopic keratoconjunctivitis, but most often, its pathology is due to direct cytotoxic effect of chemical mediators, such as eosinophil proteins, rarely results from mechanical or immunological pathways. PMID- 17417156 TI - [Different clinical forms of conjunctival allergy]. AB - Allergic conjunctivitis is a frequent disorder, benign in most cases and often associated with other manifestations of atopy such as rhinitis or asthma. This disorder is increasing steadily because of the influence of environmental factors such as pollution, smoking, etc. Allergic conjunctivitis can be broken down into five clinical forms: (1) seasonal and intra-annual conjunctivitis, (2) vernal keratoconjunctivitis, (3) atopic keratoconjunctivitis, (4) gigantopapillary conjunctivitis, and (5) contact allergies (palpebral conjunctival form). Analysis of the patterns and signs of the disorder as well as the circumstances surrounding its manifestation and an allergy workup generally facilitate eliminating the allergen responsible once it has been identified. PMID- 17417157 TI - [Childhood allergic conjunctivitis: the allergist's point of view]. AB - Allergic conjunctivitis is one of the most frequent reasons for a child's consultation with an ophthalmologist. Once the diagnosis of conjunctivitis is made and the clinical form is clearly established, the search for a cause is the most complicated step. Consultation with an allergist is necessary, in particular when questioning the patient points toward an allergic cause or brings up conditions suggestive of atopic disease. Ocular allergy can follow a type I hypersensitivity, i.e., mediated by IgE: most frequently this is acute and chronic allergic conjunctivitis. Demonstrating the allergen requires prick tests, which are easy to carry out and painless; their sensitivity is currently satisfactory. In vitro tests can complete the workup. In case of doubt on the allergen responsability for allergy, an allergenic provocation test can confirm or refute the allergen's involvement. Ocular allergy can also stem from a type IV mechanism, i.e., a cell-mediated mechanism such as in contact allergy involving different topical substances. In these cases, the clinical aspect and the patient's history may require consultation with a dermatologist-allergist for patch tests. To adapt practices when encountering conjunctivitis in a child, the ophthalmologist should be familiar with the allergens that may set off a conjunctivitis episode. Knowledge of the mechanisms at play also helps direct the examination toward allergy or another cause. In all these steps, collaboration between the ophthalmologist and the allergist is indispensable. PMID- 17417158 TI - [Conjunctival provocation test]. AB - The conjunctival provocation test (CPT) is a human model of ocular allergy that has been used to study the ocular response to allergenic stimuli and to evaluate antiallergic therapy. Considerable useful information has been gained on the ocular allergic response and drug efficacy using the conjunctival provocation test and naturally occurring seasonal allergic conjunctivitis. The safety and the efficacy of these CPTs has resulted in many authors using this tool in the etiologic diagnosis of allergic conjunctivitis. This article summarizes the possibilities of these CPTs as a tool for the study, evaluation, diagnosis, and therapeutic follow-up of allergic conjunctivitis. PMID- 17417159 TI - [Conditions bordering on allergy]. AB - Chronic allergic conjunctivitis constitutes a complex ocular surface disease involving many mechanisms, extending well beyond the simple field of mast cells and IgE. Lacrimal film, the eyelids, the environment, especially iatrogenic in origin, closely interact with each other and involve many cell systems such as goblet cells, eosinophils, and lymphocytes. It is therefore imperative to reach a better understanding of the mechanisms associated and eliminate confounding pathologies that may mimic allergic conjunctivitis. Dry eye syndrome or rosacea may be very close to chronic allergic diseases, and long-term use of preservative containing eyedrops may result in inflammatory reactions that may be very difficult to discriminate from a primarily allergic disease. PMID- 17417160 TI - [Clinical-tomographic correlation of the iridocorneal opening following laser iridotomy]. PMID- 17417161 TI - [Risk factors for microbial keratitis]. PMID- 17417162 TI - [Case reports, a relic of the past?]. PMID- 17417163 TI - [Management of lung cancer in developing countries]. PMID- 17417164 TI - [Sleep: the breakthrough]. PMID- 17417165 TI - [Habitual snoring. Prevalence and risk factors in a sample of the French male population]. AB - INTRODUCTION: As there are important differences in the prevalence of snoring in the literature we have studied this prevalence and the risk factors in a sample of the active male population of Lorraine. METHODS: 850 men, aged 22 to 66 tears, volunteered for the study and completed a questionnaire on the frequency of snoring, their lifestyle and their personal and family histories. The usual anthropomorphic measurements were made together with a non-invasive examination of the upper airways. RESULTS: The prevalence of habitual snoring was 34.6%, increasing with age, weight and the derived indices. We identified, by logistic regression, the factors independently associated with habitual snoring in univariate analysis. These were age, weight, nocturnal nasal obstruction, a medical diagnosis of obstructive sleep apnoea, and hypertrophy of the soft palate and uvula. There was evidence that height had a protective effect but exercise activity did not appear to be significant. CONCLUSION: The 35% prevalence of habitual snoring in our sample of middle aged men is similar to other studies in the literature using the same methods. The present study confirmed that age, weight, girth, nasal obstruction and nasopharyngeal abnormalities are risk factors. The protective effect of height was a new finding not identified in previous studies. PMID- 17417166 TI - [General practice management of obstructive sleep apnea in the Midi-Pyrenees]. AB - INTRODUCTION: Obstructive sleep apnoea syndrome (OSAS) is a significant public health problem. In the French care system general practitioners have an important role to play in its management especially in the field of early diagnosis. METHODS: The first part of this work is a descriptive case-control study looking at health resource usage of OSAS patients treated with continuous positive airway pressure (CPAP) in the MidiPyrenees area. The second part is a descriptive study which involved by interviewing 100 general practitioners settled in the area in order to evaluate their knowledge and their practice regarding OSAS. Results were compared with a previous study (1) performed in 2002 with similar methodology. RESULTS: Our results confirm that OSAS remains to a large extent undiagnosed (mean OSAS patients was less than 6 per physician) and underline the close connection between OSAS and cardiovascular diseases. A low degree of knowledge concerning the symptoms, main complications and treatment of OSAS was observed in this group of physicians. There was, however a relative improvement in the answers compared to the study performed in 2002. Two or more complications were mentioned by 67% of practitioners (33% in 2002) and only 13% of them were unable to identify at least one possible complication of OSAS (50% in 2002). CONCLUSION: This study confirms the impact on health of OSAS, characterized by a higher consumption of various medications than in non OSAS patients. Knowledge of OSAS remains low in general practitioner population in France. An important effort has to be made to improve learning of this disease by continuous medical training and during initial medical course. PMID- 17417167 TI - [Pulmonary infections with Mycobacterium xenopi in patients without HIV infection]. AB - OBJECTIVE: To determine the incidence, clinical characteristics, microbiological features and outcome of Mycobacterium xenopi infections in patients attending a university hospital. METHODS: We reviewed the files of HIV-seronegative patients meeting ATS criteria for M. xenopi pulmonary infection between 1993 and 2004. RESULTS: Ten patients were studied (7 men, 60+/-27 years). All but one had underlying chronic health disorders (chronic lung disease, cancer, alcoholism, systemic steroid therapy). The clinical and radiological findings were those associated with tuberculosis. Acid-fast bacilli were detected by direct examination in 9 cases, and antituberculous treatment prescribed in 8 patients. Specific treatment was started an average of 60+/-25 days after sampling, and generally combined a fluoroquinolone, clarithromycin and rifampicin, with or without ethambutol, for a mean of 11.4 months (1-37 months). Five patients had surgical excision (diagnostic in 1 case). Four patients died of their underlying disease. Two patients recovered with antibiotics alone and three with antibiotics and surgery. One patient was lost to follow-up after five months. CONCLUSION: Pulmonary infection by M. xenopi is rare in HIV-seronegative patients. The prognosis depends mainly on the patient's underlying health status. Surgery is an important component of treatment. PMID- 17417168 TI - [Prevalence of symptoms of sleep apnea syndrome. Study of a French middle-aged population]. AB - OBJECTIVE: To study the prevalence of symptoms of sleep apnoea syndrome (SAS) in a large French middle-aged population and to establish what proportion have symptoms that justify further investigation with a sleep study. METHODS: We performed a cross-sectional study of 2,195 men and 2,247 women, 33 to 69 year old (DESIR. cohort) recording responses to a self-administered "sleep" questionnaire and a general questionnaire including socio-economic characteristics and lifestyle factors. RESULTS: The prevalence of symptoms in men and women were respectively: snoring frequently (28%, 14%), frequent daytime sleepiness (14%, 18%) and frequent apnoeas (5%, 2%). Overall, 8.5% of men and 6.3% of women reported a pattern of symptoms suggestive of OSA, as they snored and had daytime sleepiness and/or apnoeas. This pattern was associated, for both sexes, with age, body mass index and after adjustment on these two factors, to a mediocre self reported health status and treatment with benzodiazepines or other sedatives. For men only, the OSA pattern of symptoms was also associated with, hypertension, alcohol consumption and smoking. CONCLUSION: Snoring, daytime hypersomnolence and witnessed apnoeas are symptoms frequently observed in the general population. Subjects with a combination of these abnormalities suggesting a high probability of sleep apnoea syndrome and in whom a sleep study is warranted represent 7.5% of the adult population. PMID- 17417169 TI - [Changes in inspiratory capacity during acute respiratory failure in COPD patients]. AB - INTRODUCTION: Inspiratory Capacity (IC), which reflects dynamic pulmonary hyperinflation, correlates with outcome in moderate exacerbation of COPD. Whether this is also true in COPD with acute respiratory failure (ARF) has not been studied. METHODS: A prospective multicenter assessment of IC measurement feasibility, reliability, time-course and relationship to outcome in COPD with ARF was conducted. Dyspnea (visual analogue scale) and IC were repeatedly measured. Outcome was classified as not favourable (death or intubation or non invasive ventilation increased or patient referred to ICU from respiratory ward) or favourable (none of the above criteria). RESULTS: Fifty patients were included and 48 analysed. IC measurement was possible in all but one patient and its coefficient of variation was 9+/-8%. Between inclusion into the study and discharge, IC increased from 39.9+/-15.5 to 50.2+/-14.5% pred (p<0.001) and dyspnea declined from 48+/-23 to 33+/-22 mm (p<0.001). Inclusion IC was not different on average between patients with or without favourable outcome. CONCLUSION: In COPD patients with ARF, IC measurement at bedside was feasible and reproducible. IC was low at entry and increased over time from inclusion to discharge tending to correlate with patient outcome. PMID- 17417170 TI - [Which definition to use when defining airflow obstruction?]. AB - INTRODUCTION: There is no clear consensus as to what constitutes an obstructive ventilatory defect (OVD). According to the American Thoracic Society and European Respiratory Society, it is defined as being when the ratio of the forced expiratory volume (FEV1) and the slow expiratory vital capacity (VC) is below the lower limit of normal (LLN). According to the Global initiative for chronic Obstructive Lung Disease and the British Thoracic Society, it is an FEV1/forced expiratory vital capacity (FVC)<0.70 and an FEV1<80%. In addition, in daily practice, the OVD is diagnosed by a "Fixed ratio" FEV1/FVC<0.70 or30/hr] in 8 patients. Those having AHI<30 hr or a failure of HRP (5 patients) were studied by full polysomnography in the sleep laboratory (PSG). RESULTS: In 52 patients the AHI obtained by HRP and analysed manually correlated weakly with that obtained by PSG (n=52; p<0.001; r=0.36). The AHI-PSG was 27.1+/-2.8/hr and the AHI-HRP was 12+/-0.9/hr. The mean difference (HRP-PSG) was 15.1+/-37.5/hr with poor concordance. A better cut off value in terms of efficacy of HRP was an AHI of 10/hr, with sensitivity of 61.4% and a specificity of 100%. CONCLUSION: A negative result by HRP does not exclude OSAS and full PSG is required in patients suspected of having this condition. PMID- 17417172 TI - [Catamenial hemoptysis during hormone replacement treatment]. AB - INTRODUCTION: Catamenial haemoptysis is a rare clinical entity resulting from the presence of ectopic intra pulmonary endometrial tissue, either parenchymatous or endobronchial. The main diagnostic criterion is the periodic character of the haemoptysis which is synchronous with menstruation. CASE REPORT: The authors report a case of catamenial haemoptysis due to endobronchial endometriosis in a 46 year old menopausal woman receiving hormone replacement treatment (HRT). She presented with 3 episodes of haemoptysis synchronous with the first days of her menstrual cycle. A thoracic CT scan showed ground glass lesions with micronodulation. Bronchoscopy showed violacious lesions bleeding on contact. The endobronchial and CT abnormalities had disappeared by day 5. After withdrawal of the HRT the haemoptysis did not recur during a follow-up of 2 years. CONCLUSION: Endobronchial endometrioisis remains a rare occurrence. This is the first case reported in a menopausal woman with artificial cycles receiving hormone replacement therapy. PMID- 17417173 TI - [Serious complication of radiofrequency treatment of inoperable bronchial carcinoma]. AB - INTRODUCTION: Surgery remains the only curative treatment for primary non-small cell bronchial carcinoma. It is mainly appropriate for small, localised tumours. Some patients have contra-indications to surgery and radiofrequency offers a minimally invasive alternative with few complications. It is performed under general anaesthesia by a percutaneous approach. The main complications are mechanical, primarily pneumothorax, and infections are uncommon and generally mild. CASE REPORT: We report the case of a man treated by radiofrequency for a small bronchial carcinoma. The procedure was rapidly complicated by infection of the area treated, spreading throughout both lung fields and requiring intensive and prolonged antibiotic treatment. Resolution of the infection was slow despite appropriate treatment. The rapid onset and spread are explained by the immunosuppressed state of the patient. CONCLUSION: Radiofrequency is a recent treatment for bronchial carcinoma that is developing rapidly. Though it is associated with low morbidity and mortality the possibility of potentially fatal infective complications in certain patients should be recognised. PMID- 17417174 TI - [Pulmonary actinomycosis with chest wall fistula formation]. AB - INTRODUCTION: Actinomycosis is a suppurative infection caused by bacteria of the genus actinomyces. It is a rare cause of pulmonary infection and can be difficult to diagnose as its presentation may mimic cancer or tuberculosis. In the absence of treatment of pulmonary lesions fistulae can develop. We report a case of pulmonary actinomycosis complicated by a chest wall fistula that was managed by the thoracic surgery unit at Chu Ibn Sina de Rabat. CASE REPORT: A 45 year old man who was previously fit and well presented with a left sided chest wall swelling with cutaneous fistulae. Pulmonary auscultation revealed evidence of left apical consolidation. Otherwise clinical assessment was unremarkable. Plain chest radiology revealed left apical opacification. CT scanning revealed a left pulmonary mass and percutaneous biopsy confirmed the diagnosis of actinomycosis. The patient was started on treatment and experienced a clinical and radiological improvement sustained at one year's follow up. CONCLUSION: We review the clinical and radiological characteristics of this condition as well as diagnostic difficulties, histological findings, therapeutic options and its natural history. PMID- 17417175 TI - [Nocardial pulmonary infection]. AB - INTRODUCTION: Nocardial pneumonias are due to a genus of aerobic, filamentous, partly acid-alcohol fast, mainly Gram positive, actinomycetes. CASE REPORT: We report here two cases of nocardial pneumonia. The first was a 62 year old man with a history of fludaribine treatment and bone marrow transplant for lymphocytic leukaemia. During the investigation of pyrexia evidence of N. farcinica infection was found in the bronchial secretions. The second case was a man of 61 receiving long term corticosteroids and cytotoxic chemotherapy. Investigation of a pneumonia with pleural effusion found evidence, on culture of blood and pleural fluid, of disseminated infection with N. nova (cerebral, pleural, pulmonary and splenic). CONCLUSION: Nocardiosis is a rare cause of pneumonia mainly occurring in immuno-compromised adults (corticosteroid therapy, HIV infection, transplantation, cancer or leukaemia). It should be suspected in the presence of pleuro-pulmonary symptoms associated with neurological and cutaneous signs, general deterioration and weight loss. The microbiology laboratory should be advised of this eventuality as soon as possible in order to optimise the search for the organism. PMID- 17417176 TI - [Pulmonary arterial hypertension due to tumor emboli]. AB - INTRODUCTION: Pulmonary arterial hypertension (PAH) is rare in the presence of malignancy and tumour embolisation is one of several possible pathological mechanisms. CASE REPORTS: We report our experience of 5 clinical cases and undertake a literature revue of the pathophysiological mechanisms and of the possible diagnostic and therapeutic approaches. CONCLUSIONS: Neoplastic PAH due to tumour micro-emboli is rare and the diagnosis difficult to establish. Cytological examination of pulmonary arterial blood could allow early institution of appropriate chemotherapy and lead to an improvement in the grave prognosis of this condition. PMID- 17417177 TI - [Pulmonary artery stump thrombosis following pneumonectomy]. AB - INTRODUCTION: Pulmonary artery stump thrombosis has been reported after pneumonectomy. The prevalence of risk factors for and outcome associated with this condition remain to be clearly evaluated. CASE REPORT: We describe three cases of pulmonary artery stump thrombosis occurring after pneumonectomy. No pulmonary embolism was observed despite the absence of anticoagulation therapy. The prevalence of arterial stump thrombosis after pneumonectomy has been estimated as 12%. It is often detected by CT scanning performed systematically as part of the routine follow up of lung cancer. Thrombus was identified as a filling defect with concave or convex margins, located at the bottom of the stump. DISCUSSION: Only one case of pulmonary embolism associated with pulmonary artery stump thrombosis has been described. The absence of complications associated with thrombus in this site suggests that anticoagulant therapy is not likely to be of benefit. PMID- 17417178 TI - [Multiple cavitating pulmonary nodules in a virology laboratory assistant]. PMID- 17417180 TI - [Diagnostic approach to infectious lung diseases in patients immunocompromised by HIV]. PMID- 17417181 TI - [Community-acquired respiratory infections in non-immunocompromised adults]. PMID- 17417182 TI - [Acute community-acquired pneumonia in non-immunocompromised infants]. PMID- 17417183 TI - [Research training in respiratory medicine, 2006]. PMID- 17417218 TI - [Abstracts of the Neurology Meeting of the French Speaking Community, Paris, France, 11-14 April 2007]. PMID- 17417600 TI - Polymorphisms of chemokine and chemokine receptor genes in idiopathic immune mediated posterior segment uveitis. AB - PURPOSE: Chemokines are important inflammatory mediators that play a crucial role in uveitis. Polymorphisms in chemokine genes can alter the expression of these genes in the inflammatory cells, which, in turn, can affect the clinical phenotype of the disease. The purpose of this study was to identify polymorphisms in chemokine genes that can predict visual outcome in patients with immune mediated posterior segment uveitis. METHODS: This is a case-control study of 141 Caucasians with idiopathic immune-mediated posterior segment uveitis and 282 controls matched by age and ethnicity. Six polymorphisms in four genes, (MCP-1 2518A/G, RANTES-403G/A, RANTES-28C/G, CCR2 V64I, CCR5-59029G/A, and CCR5 32 bp deletion) were analyzed by sequence specific primers polymerase chain reaction. RESULTS: Patients with G allele at MCP-1-2581 developed the disease at an early age as compared to patients with A allele corrected p value pc=0.003. Also patients with A allele at RANTES-403 position developed less severe disease and had better visual outcome when compared with patients with G allele (pc=0.02) Final visual acuity after 18 months was better in patients with 32 bp deletion of the CCR5 gene and in patients with the CCR2 wild-type genotype pc=0.02 and pc=0.04, respectively. Patients with the CCR2 64I allele also had a higher risk of developing an elevated intraocular pressure as compared to patients with the wild-type genotype (pc=0.007). CONCLUSIONS: Though the utility for prediction of disease susceptibility of the studied polymorphisms in chemokine genes is in general not robust, we have found that polymorphisms in chemokine genes can influence the outcome of patients with idiopathic immune-mediated posterior segment uveitis. These associations require further analysis in other groups of patients. PMID- 17417601 TI - Elevated insulin signaling disrupts the growth and differentiation pattern of the mouse lens. AB - PURPOSE: Insulin and insulin-like growth factors (IGFs) are putative regulators of cell proliferation and differentiation during lens development. Transgenic mice that overexpress IGF-1 in the lens have been previously described. To further understand the ocular functions of this growth factor family, the in vivo effects of insulin expression on lens development were investigated using transgenic mice. METHODS: Expression of insulin receptor (IR) and IGF-1 receptor (IGF-1R) in mouse lens was examined by reverse-transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization. Transgenic mice that overexpress insulin in the lens were generated using two different promoters: a fiber-cell specific alphaA-crystallin (alphaA) promoter and a modified alphaA-promoter linked to the chicken delta1-crystallin enhancer (called the deltaenalphaA promoter). The deltaenalphaA promoter is active in both lens epithelial and fiber cells. The lens phenotypes were analyzed by histology and immunohistochemistry. Protein expression was examined by western blotting. RESULTS: Normal mouse lenses express both the insulin receptor (IR) and the IGF-1 receptor (IGF-1R), and their expression is highest at the lens periphery where the germinative and transitional zones are located. In transgenic mice, insulin expression in the lens induced cataract formation. The severity of the cataracts reflected the level of transgene expression, independent of the type of promoter used. In severely affected families, the spherical shape of the lens was altered and the lenses were smaller than normal. Histological analysis showed no evidence of premature differentiation of the anterior epithelial cells. In contrast to the IGF-1 mice, insulin transgenic mice exhibited an anterior shift in the location of the germinative and transitional zones, leading to a reduction of the lens epithelial compartment. Additional alterations included expansion of the lens transitional zone, variable nuclear positioning in the lens bow region, and inhibition of fiber cell denucleation and terminal differentiation. CONCLUSIONS: Elevated intraocular insulin does not enhance proliferation nor induce differentiation of mouse lens epithelial cells. Since an increase in IGF-1 causes a posterior shift of the lens geminative and transitional zones, while an increase in insulin causes an anterior shift of these zones, our results suggest that these two growth factors may work together to control the location of this structural domain during normal lens development. Our data also suggest that increased insulin-signaling activity in the lens can antagonize the endogenous signals that are responsible for fiber cell maturation and terminal differentiation. PMID- 17417602 TI - Phospholipase A2 in chamber angle of normal eyes and patients with primary open angle glaucoma and exfoliation glaucoma. AB - PURPOSE: Phospholipase A2 (PLA2) is a growing family of lipolytic enzymes that play a key role in various biological processes including general lipid metabolism, membrane homeostasis, and in diseases such as atherosclerosis, arthritis, and acute pancreatitis. Oxidative stress as well as inflammation may be associated with glaucoma pathogenesis. Therefore, our aim was to examine the expression of group IIA secretory PLA2 (sPLA2-IIA), group V secretory PLA2 (sPLA2 V), calcium-independent PLA2 (iPLA2), and cytosolic PLA2 (cPLA2) type in the trabecular meshwork (TM) and the canal of Schlemm in normal eyes and in juxtacanalicular tissue samples from patients with primary open angle glaucoma (POAG) or exfoliation glaucoma (ExG). METHODS: TM tissues were isolated from healthy donor eyes for corneal transplantation. Specimens of inner wall of the Schlemm's canal and the juxtacanalicular tissue were collected during deep sclerectomy from the eyes of patients who had POAG or ExG. Antibodies against PLA2s (sPLA2-IIA, sPLA2-V, iPLA2, and cPLA2) and a standard immunohistochemical procedure were used for the analysis. Quantification of immunoreactions was provided using a Photoshop-based image analysis. Double-staining immunofluorescence of macrophages and sPLA2-IIA was performed by using confocal microscopy. RESULTS: sPLA2-IIA was not present in normal TM. In contrast, sPLA2 IIA levels were significantly higher in glaucoma patients than in controls. Furthermore, sPLA2-IIA expression was much higher in POAG when compared to ExG. iPLA2 was found to predominate in normal human TM, and it demonstrated strong labeling in the uveal and corneoscleral meshwork. The staining of juxtacanalicular meshwork was only moderate in density. In contrast, expression of the enzyme was significantly decreased in glaucoma patients, especially in ExG, when compared to normal controls or to POAG. In addition, strong regional differences were detected in sPLA2-IIA and iPLA2 levels in POAG, whereas immunostaining of these enzymes was much lower and rather uniform throughout ExG sample. In POAG, sPLA2-IIA staining was restricted to certain parts of the trabecular samples where sPLA2-IIA positive macrophages were also present. Immunostaining of sPLA2-V or cPLA2 was low, and no significant changes were found in levels of these enzymes between normal and glaucomatous samples. CONCLUSIONS: sPLA2-IIA, an oxidative stress marker in atherosclerosis, is overexpressed especially in POAG. This result supports the hypothesis that oxidative stress may play a significant role in the pathogenesis of POAG. In ExG, a dramatic decrease in the expression level of iPLA2, a housekeeping enzyme in phospholipid remodeling, may indicate imbalance in phospholipid turnover and also inhibition of normal physiological functions in the TM. These findings may contribute to understanding the pathogenesis of POAG and ExG and may be important for the development of novel therapeutic strategies to different glaucomas. PMID- 17417603 TI - Focal adhesion kinase (FAK) expression and activation during lens development. AB - PURPOSE: Regulation of lens development involves an intricate interplay between growth factor (e.g. FGF and TGFbeta) and extracellular matrix (ECM) signaling pathways. Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that plays key roles in transmitting ECM signals by integrins. In this study, we delineated patterns of FAK expression and tyrosine phosphorylation (Y397) in the developing lens and investigated its regulation by FGF2. We also examined FAK expression and activation during disrupted fiber differentiation in mice expressing a dominant negative TGFbeta receptor. METHODS: FAK expression and activation (phosphorylation on Y397) was studied in embryonic and postnatal rodent lenses by in situ hybridization, immunofluorescence, and western blotting. Rat lens explants were used to investigate the effects of FGF2 on FAK expression and activation. Immunofluorescence and western blotting were used to examine FAK expression and phosphorylation in transgenic mice that express a dominant negative TGFbeta receptor. RESULTS: FAK is widely expressed and phosphorylated during embryonic stages of lens morphogenesis and differentiation. However, in postnatal lenses its expression and activation becomes restricted to the posterior germinative zone and the transitional zone at the lens equator. While both NH2- and COOH-terminal antibodies revealed cytoplasmic and membrane associated staining in lens cells, the NH2-terminal antibody also showed FAK was present in fiber cell nuclei. In vitro, FAK expression and phosphorylation on Y397 were increased by concentrations of FGF2 that initiate lens epithelial cell migration (10 ng/ml) and differentiation (50 ng/ml) but not proliferation (5 ng/ml). Moreover, reactivity for Y397 phosphorylated FAK is prominent in the nuclei of differentiating fibers both in vivo and in vitro. Disruption of TGFbeta like signals by ectopic expression of a dominant-negative TGFbeta receptor (TbetaRII(D/N)) results in abnormal lens fiber differentiation in transgenic mice. While FAK expression is initiated normally in the posterior germinative zone of TbetaRII(D/N) transgenic lenses, as fiber differentiation proceeds, FAK becomes localized to a perinuclear compartment, decreases its association with the cytoskeleton and is poorly phosphorylated on Y(397). CONCLUSIONS: FAK is widely expressed and activated during early lens morphogenesis. During secondary lens fiber differentiation, FAK is expressed and phosphorylated on Y397 as epithelial cells exit the cell cycle, initiate migration at the equator, and undergo differentiation in the transitional zone. During terminal fiber differentiation an NH2-terminal fragment of FAK, including Y397, is translocated to the nucleus. The expression, activation, and nuclear localization of FAK are regulated, at least partly, by FGF2. FAK activity and subcellular localization are also modulated by TGFbeta-like signals. In fiber cells of TbetaRII(D/N) transgenic lenses, FAK is abnormally retained in a perinuclear compartment, loses its association with the cytoskeleton, and is poorly phosphorylated. These data suggest that integrin signaling via FAK plays important roles during lens differentiation, mediated by FGFs and TGFbeta-superfamily signals. PMID- 17417604 TI - Cloning and characterization of canine PAX6 and evaluation as a candidate gene in a canine model of aniridia. AB - PURPOSE: Mutations in PAX6 cause human aniridia. The small eye (sey) mouse represents an animal model for aniridia. However, no large animal model currently exists. We cloned and characterized canine PAX6, and evaluated PAX6 for causal associations with inherited aniridia in dogs. METHODS: Canine PAX6 was cloned from a canine retinal cDNA library using primers designed from human and mouse PAX6 consensus sequences. An RH3000 radiation hybrid panel was used to localize PAX6 within the canine genome. Genomic DNA was extracted from whole blood of dogs with inherited aniridia, and association testing was performed using markers on CFA18. Fourteen PAX6 exons were sequenced and scanned for mutations, and a Southern blot was used to test for large deletions. RESULTS: Like the human gene, canine PAX6 has 13 exons and 12 introns, plus an alternatively spliced exon (5a). PAX6 nucleotide and amino acid sequences were highly conserved between dog, human, and mouse. The canine PAX6 cDNA sequence determined in this study spans 2 large gaps present in the current canine genomic sequence. Radiation hybrid mapping placed canine PAX6 on CFA18 in a region with synteny to HSA11p13. Exon scanning revealed single nucleotide polymorphisms, but no pathological mutations, and Southern blot analysis revealed no differences between normal and affected animals. CONCLUSIONS: Canine PAX6 was cloned and characterized, and results provide sequence information for gaps in the current canine genome sequence. Canine PAX6 nucleotide and amino acid sequences, as well as gene organization and map location, were highly homologous with that of the human gene. PAX6 was evaluated in dogs with an inherited form of aniridia, and sequence analysis indicated no pathological mutations in the coding regions or splice sites of aniridia-affected dogs, and Southern blot analysis showed no large deletions. PMID- 17417605 TI - Ca2+ channels in retinal pigment epithelial cells regulate vascular endothelial growth factor secretion rates in health and disease. AB - PURPOSE: Choroidal neovascularization (CNV) is the most severe complication in age-related macular degeneration. The major angiogenic factor involved is vascular endothelial growth factor (VEGF) secreted by the retinal pigment epithelium (RPE). Since RPE cells express neuroendocrine L-type Ca2+ channels we investigated their involvement in VEGF secretion in normal RPE cells and RPE cells from patients with CNV. METHODS: Freshly isolated and cultured RPE cells were studied using the patch-clamp technique and ELISA-based secretion assays. RESULTS: Both freshly isolated and cultured cells showed whole-cell Ba2+ currents with properties of L-type Ca2+ currents: high activation threshold, sensitivity to dihydropyridines (10 muM nifedipine) and slow inactivation. VEGF-A secretion was elevated by BayK8644 (10 microM) or basic fibroblast growth factor (bFGF, 10 ng/ml), both of which are able to activate L-type channels. Cells from CNV tissue also showed nifedipine-sensitive Ba2+ currents, which displayed a voltage dependent activation at more negative potentials, faster inactivation and changed regulation by tyrosine kinase pp60(c-src). The CNV RPE cells showed higher VEGF secretion rates which were reduced by nifedipine. CONCLUSIONS: Thus, L-type Ca2+ channels in normal RPE cells regulate the secretion of VEGF. RPE cells from eyes with CNV maintain a VEGF secretion regulated by nifedipine-sensitve Ca2+ channels which might be of importance for the development of CNV. PMID- 17417606 TI - Adenoviral gene transfer of bioactive TGFbeta1 to the rodent eye as a novel model for anterior subcapsular cataract. AB - PURPOSE: To produce a gene-transfer model of rodent anterior subcapsular cataracts (ASC) using a replication-deficient, adenoviral vector containing active TGFbeta1. Establishment of this model will be important for further investigations of TGFbeta-induced signaling cascades in ASC. METHODS: Adenovirus containing the transgene for active TGFbeta1 (AdTGFbeta1), beta-galactosidase (AdLacZ), green fluorescent protein (AdGFP) or no transgene (AdDL) was injected into the anterior chamber of C57Bl/6, Smad3 WT and Smad3 KO mice. Four and 21 days after injection, animals were enucleated and eyes were processed and examined by routine histology. Immunolocalization of markers indicative of epithelial to mesenchymal transition (EMT), fibrosis, proliferation and apoptosis was also carried out. RESULTS: By day 4, treatment with AdLacZ demonstrated transgene expression in multiple structures of the anterior chamber including the lens epithelium. In contrast to AdDL, treatment with AdTGFbeta1 produced alphaSMA positive subcapsular plaques in all three groups of mice, which shared features reminiscent of human ASC. At day 21, plaques remained alphaSMA-positive and extensive extracellular matrix deposition was observed. The AdTGFbeta1 model was further employed in Smad3 deficient mice and this resulted in the development of small ASC. CONCLUSIONS: Gene transfer of active TGFbeta1 using an adenoviral vector produced cataractous plaques four days postinjection, which were found to develop independent of functional Smad3. PMID- 17417607 TI - Identification of three novel NHS mutations in families with Nance-Horan syndrome. AB - PURPOSE: Nance-Horan Syndrome (NHS) is an infrequent and often overlooked X linked disorder characterized by dense congenital cataracts, microphthalmia, and dental abnormalities. The syndrome is caused by mutations in the NHS gene, whose function is not known. The purpose of this study was to identify the frequency and distribution of NHS gene mutations and compare genotype with Nance-Horan phenotype in five North American NHS families. METHODS: Genomic DNA was isolated from white blood cells from NHS patients and family members. The NHS gene coding region and its splice site donor and acceptor regions were amplified from genomic DNA by PCR, and the amplicons were sequenced directly. RESULTS: We identified three unique NHS coding region mutations in these NHS families. CONCLUSIONS: This report extends the number of unique identified NHS mutations to 14. PMID- 17417608 TI - Phenotypes, distribution, and morphological features of antigen-presenting cells in the murine cornea following intravitreal injection. AB - PURPOSE: To study the phenotypes, distribution, and morphologies of different antigen-presenting cells (APCs) in the murine cornea. METHODS: Intravitreal injection of fluorescently tagged ovalbumin (OVA) or antibodies to MHC-II (I A(d)), F4/80, CD11c, B7-1, and B7-2 was performed to label cells in the murine cornea. Light and transmission electron microscopy were used to examine corneal histology. Intravital microscopy, epifluorescence microscopy, and confocal microscopy were used to evaluate the labeled cells. In vitro staining was performed to validate the in vivo staining and localize the labeled cells. Three dimensional rotatable images were taken to evaluate relationships between two differently labeled cells. RESULTS: Histological examination revealed no observable change in the cornea following intravitreal injection. In vivo staining showed that OVA+ cells and cells positive for MHC-II, F4/80, CD11c, B7 1, or B7-2 were noted throughout the cornea with a decreasing density from limbus toward the central cornea. Two populations with distinct morphological features were identified among these APCs. Labeled cells were found beneath the epithelium or in the shallow stroma in the central and paracentral cornea, but in all layers in the peripheral cornea. A number of F4/80+ and CD11c+ cells were also positive for OVA, MHC-II, B7-1, or B7-2. Rotatable images showed a close contact between two differently labeled cells. CONCLUSIONS: Intravitreal injection of labeled antibodies can be adapted to visualize labeled cells in the cornea. APCs with distinct morphologies, phenotypes, and distribution may contribute to the immunologically privileged feature of the cornea. PMID- 17417609 TI - Investigation of founder effects for the Thr377Met Myocilin mutation in glaucoma families from differing ethnic backgrounds. AB - PURPOSE: The aim of this study was to determine if there is a common founder for the Thr377Met myocilin mutation in primary open angle glaucoma (POAG) families with various ethnic backgrounds. METHODS: Genomic DNA of 24 POAG-affected individuals from nine pedigrees with the Thr377Met mutation and 104 unaffected family members was genotyped with six microsatellite markers and four single nucleotide polymorphisms. The families were from Greece, India, Finland, the USA, and Australia. To assess the degree of linkage disequilibrium across MYOC in the general population we also investigated data generated from the HapMap consortium. RESULTS: Three distinct haplotypes associated with the Thr377Met myocilin mutation were identified. The families from the USA and Greece, as well as the three Australian families originating from Greece and the former Yugoslavian Republic of Macedonia had one common haplotype. Interestingly, however, HapMap data suggest that linkage disequilibrium across MYOC was not strong. CONCLUSIONS: The Thr377Met myocilin mutation has arisen at least three separate times. Evidence for genetic founder effects in this prevalent age related, yet heterogeneous, disease has important implications for future gene identification strategies. PMID- 17417610 TI - Inhibition by brimonidine of forskolin-induced nitric oxide synthase expression in human ciliary bodies in vitro. AB - PURPOSE: To investigate the mRNA and protein expression of nitric oxide synthase (NOS) in human ciliary bodies in vitro. The effect of the adenylcyclase activator forskolin and/or the alpha2-adrenergic agonist brimonidine (an ocular hypotensive agent that inhibits aqueous humor formation) on NOS mRNA or protein expression was also studied. METHODS: Frozen human ciliary bodies obtained from local eye bank were thawed and incubated with 0.1 mM forskolin for 24 h in the absence or in the presence of 100 muM brimonidine. The mRNA and protein expression of three NOS isoforms (neuronal NOS or nNOS, inducible NOS or iNOS, endothelial NOS or eNOS) were assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively. RESULTS: mRNA and protein expression of three NOS isoforms were detected in human ciliary bodies. Forskolin significantly up-regulated the mRNA and protein expression of nNOS, but not that of iNOS or of eNOS. In the presence of brimonidine, the forskolin-induced up-regulation of nNOS mRNA or protein expression was significantly inhibited. CONCLUSIONS: In human ciliary body (where aqueous humor is produced), brimonidine inhibits the up regulation of nNOS expression induced by forskolin. PMID- 17417611 TI - Myocilin mutations among primary open angle glaucoma patients of Kanyakumari district, South India. AB - PURPOSE: Glaucoma can be defined as optic neuropathy leading to irreversible blindness if not treated in time. Primary open angle glaucoma (POAG) is the most common form of glaucoma. The myocilin (MYOC) gene has been found to mutate in both sporadic and familial cases of POAG worldwide. About 90% of these mutations have been seen to cluster at exon III of the gene. There are documented reports of mutations in the MYOC gene among POAG patients from different parts of India. The southernmost tip of the Indian subcontinent (Kanyakumari district) has remained isolated from all these studies. The aim of this study was to indicate or rule out the disease causative role of the MYOC gene mutations in these patients by screening the MYOC gene for mutations among POAG patients of the Kanyakumari district. METHODS: One hundred POAG patients from the Kanyakumari District of South India were recruited for the study. The MYOC gene was screened using the PCR-SSCP methodology followed by DNA sequencing. The sequences were analyzed using BLAST. Secondary structures of the amino acid sequences with a variation were predicted. RESULTS: Two probable disease-causing variations (mutations), Ser331Thr and Pro370Leu, were each observed in one patient apiece. Two polymorphisms, (Tyr347Tyr and Thr325Thr) were also observed in the patients. Ser331Thr is a novel conservative change while Pro370Leu is a widely reported mutation with an associated severe disease phenotype. CONCLUSIONS: The presence of the mutations in the patients suggests the causative role of the MYOC gene among POAG patients in the Kanyakumari district of India. The mutation frequency of 2% corresponds well with the other reports from India and other countries. However, the mutation rate reported from a population in the eastern part of India was much higher. Screening of patients from different parts of India is essential to estimate the overall mutation frequency. More functional studies on the MYOC gene are required to elucidate the pathophysiology of POAG. PMID- 17417612 TI - Atomic force microscopy measurements of lens elasticity in monkey eyes. AB - PURPOSE: To demonstrate the feasibility of measuring the elasticity of intact crystalline lenses using atomic force microscopy (AFM). METHODS: AFM elasticity measurements were performed on intact lenses from 18 fresh cynomolgus monkey cadaver eyes (4-10 years old, <1 day postmortem) that had been left attached to their zonule-ciliary body-sclera framework. The eyes were prepared by bonding a plastic ring on the sclera after removal of the conjunctival, adipose, and muscle tissues. The posterior pole was sectioned, with the excess vitreous removed, and the eye's anterior section was placed on a Teflon slide to protect the posterior pole of the lens. The cornea and iris were then sectioned. The lens-zonule ciliary body-sclera section was then placed in a Petri dish filled with balanced salt solution in an AFM system designed for force measurements. Next, the central pole of the anterior surface of the intact lens was probed with the AFM cantilever tip. The recorded AFM cantilever deflection-indentation curves were used to derive force-indentation curves for the lens after factoring out the deflection of the cantilever on a hard surface. Young's modulus of the lens was calculated from the force-indentation relation using the Hertz model. RESULTS: Young's modulus was 1,720+/-880 Pa (range: 409-3,210 Pa) in the 18 cynomolgus monkey lenses. CONCLUSIONS: AFM can be used to provide measurements of the elasticity of the whole lens including the capsule. Values obtained using AFM on cynomolgus monkey lenses are similar to published values obtained using dynamic mechanical analysis on young human lenses. PMID- 17417613 TI - Three new PAX6 mutations including one causing an unusual ophthalmic phenotype associated with neurodevelopmental abnormalities. AB - PURPOSE: The PAX6 gene was first described as a candidate for human aniridia. However, PAX6 expression is not restricted to the eye and it appears to be crucial for brain development. We studied PAX6 mutations in a large spectrum of patients who presented with aniridia phenotypes, Peters' anomaly, and anterior segment malformations associated or not with neurological anomalies. METHODS: Patients and related families were ophthalmologically phenotyped, and in some cases neurologically and endocrinologically examined. We screened the PAX6 gene by direct sequencing in three groups of patients: those affected by aniridia; those with diverse ocular manifestations; and those with Peters' anomaly. Two mutations were investigated by generating crystallographic representations of the amino acid changes. RESULTS: Three novel heterozygous mutations affecting three unrelated families were identified: the g.572T>C nucleotide change, located in exon 5, and corresponding to the Leucine 46 Proline amino-acid mutation (L46P); the g.655A>G nucleotide change, located in exon 6, and corresponding to the Serine 74 Glycine amino-acid mutation (S74G); and the nucleotide deletion 579delG del, located in exon 6, which induces a frameshift mutation leading to a stop codon (V48fsX53). The L46P mutation was identified in affected patients presenting bilateral microphthalmia, cataracts, and nystagmus. The S74G mutation was found in a large family that had congenital ocular abnormalities, diverse neurological manifestations, and variable cognitive impairments. The 579delG deletion (V48fsX53) caused in the affected members of the same family bilateral aniridia associated with congenital cataract, foveal hypolasia, and nystagmus. We also detected a novel intronic nucleotide change, IVS2+9G>A (very likely a mutation) in an apparently isolated patient affected by a complex ocular phenotype, characterized primarily by a bilateral microphthalmia. Whether this nucleotide change is indeed pathogenic remains to be demonstrated. Two previously known heterozygous mutations of the PAX6 gene sequence were also detected in patients affected by aniridia: a de novo previously known nucleotide change, g.972C>T (Q179X), in exon 8, leading to a stop codon and a heterozygous g.555C>A (C40X) recurrent nonsense mutation in exon 5. No mutations were found in patients with Peters' anomaly. CONCLUSIONS: We identified three mutations associated with aniridia phenotypes (Q179X, C40X, and V48fsX53). The three other mutations reported here cause non-aniridia ocular phenotypes associated in some cases with neurological anomalies. The IVS2+9G>A nucleotide change was detected in a patient with a microphthalmia phenotype. The L46P mutation was detected in a family with microphthalmia, cataract, and nystagmus. This mutation is located in the DNA binding paired-domain and the crystallographic representations of this mutation show that this mutation may affect the helix-turn-helix motif, and as a consequence the DNA-binding properties of the resulting mutated protein. Ser74 is located in the PAX6 PD linker region, essential for DNA recognition and DNA binding, and the side chain of the Ser74 contributes to DNA recognition by the linker domain through direct contacts. Crystallographic representations show that the S74G mutation results in no side chain and therefore perturbs the DNA-binding properties of PAX6. This study highlights the severity and diversity of the consequences of PAX6 mutations that appeared to result from the complexity of the PAX6 gene structure, and the numerous possibilities for DNA binding. This study emphasizes the fact that neurodevelopmental abnormalities may be caused by PAX6 mutations. The neuro-developmental abnormalities caused by PAX6 mutations are probably still overlooked in the current clinical examinations performed throughout the world in patients affected by PAX6 mutations. PMID- 17417617 TI - Treatment of macula-on retinal detachments. PMID- 17417619 TI - Acute central retinal artery occlusion in Adamantiades-Behcet disease. PMID- 17417620 TI - Correlation between microaneurysm closure rate and reduction in macular thickness following laser photocoagulation of diabetic macular edema. AB - AIM: To correlate the microaneurysmal closure rate measured on fundus fluorescein angiography (FFA) with reduction in macular thickness observed on optical coherence tomography (OCT) following laser photocoagulation of diabetic macular edema. MATERIALS AND METHODS: A prospective observational case series. Fifty patients (50 eyes) of type II diabetes mellitus with clinically significant macular oedema (CSME) underwent focal/grid laser photocoagulation. OCT and FFA were performed at baseline and at 2 and 12 weeks following laser photocoagulation to measure the change in macular thickness and the number of leaking microaneurysms respectively. Statistical analysis was performed using paired ttest and Pearson's correlation test. RESULTS: A significant reduction in macular thickness was seen at both 2 (P=0.02) and 12 weeks (P<0.0001), most remarkably in the central 1 mm quadrant. However, microaneurysm closure was only 0.67% at 2 weeks, which increased to 89.6% by 12 weeks. The change in retinal thickness correlated significantly with the decrease in the number of leaking microaneurysms at 12 weeks (r=0.597, P<0.0001), but not at 2 weeks (r=-0.228, P=0.112). On OCT, the final reduction in retinal thickness achieved at 12 weeks from baseline correlated significantly with the initial decrease in retinal thickness at 2 weeks (r=0.66, P<0.0001). However, on FFA, the final closure rate of leaking microaneurysms at 12 weeks from baseline did not correlate with the initial closure rate at 2 weeks (r=-0.039, P=0.413). CONCLUSION: Following laser photocoagulation for CSME, an OCT at 2 weeks is more informative and better correlates with the final outcome than an FFA at 2 weeks. PMID- 17417621 TI - Vascularised epiretinal membrane associated with vasoproliferative tumour. PMID- 17417622 TI - 'Layer by layer' phacoemulsification in posterior polar cataract with pre existing posterior capsular rent. AB - PURPOSE: To evaluate a new technique of 'Layer by Layer' phacoemulsification in eyes with posterior polar cataract with pre-existing posterior capsular rent (PCR). METHODS: Layer by layer phacoemulsification was performed in eight eyes of eight patients of posterior polar cataract with pre-existing posterior capsular rent. All the patients had evidence of a posterior capsular defect showing presence of iridescent refractile lens particles in vitreous with onion skinning of central dense plaque. RESULTS: The mean age of the patients was 43.75+/-2.5 years and six patients were men. All the eyes had vertical posterior capsular defect irrespective of the site of entry of phaco probe. Vitrectomy was not required in any of the eyes. Single piece foldable acrylic intraocular lens was implanted in the bag in all the eyes. The mean preoperative best corrected visual acuity was 0.14+/-0.05, which improved to 0.77+/-0.27. CONCLUSION: Our technique of 'Layer by layer' phacoemulsification is an effective and safe technique for the management of posterior polar cataract with pre-existing PCR. PMID- 17417623 TI - Ketamine sedation during the treatment of retinopathy of prematurity. AB - AIMS: To report the use of ketamine sedation as an alternative anaesthetic method for babies undergoing treatment for retinopathy of prematurity (ROP). METHODS: All babies who underwent treatment for ROP over a 2-year period were included in this study. The babies preoperative weight, medical condition, and ventilation status was recorded. Data were collected on their ventilation status pre-, intra , and postprocedure. Any change in their cardiac or respiratory status during or in the subsequent 3 days following the treatment was noted. RESULTS: Eleven babies, 22 eyes, required treatment over this period. The procedure was well tolerated with only three babies having intraoperative complications, which all resolved spontaneously. Two babies had postoperative complications requiring additional ventilation. In no case was the procedure abandoned owing to anaesthetic complications. CONCLUSIONS: The use of ketamine sedation allows the laser to be performed in a ward setting and avoids the potential risk of general anaesthesia and inter- and intra-hospital transfer. It has been found to produce few intra- or postoperative complications for the infant, while providing satisfactory conditions for the treatment of ROP. PMID- 17417624 TI - Does an iridotomy provide protection against narrowing of the anterior chamber angle during Valsalva maneuvre in eyes with primary angle closure. AB - PURPOSE: To evaluate changes at the anterior chamber angle during Valsalva manoeuver, in eyes having primary angle closure (PAC) and a patent laser peripheral iridotomy. METHODS: Twenty-three eyes of 23 consecutive patients underwent a recording of applanation tonometry, measurement of the anterior chamber angle recess, angle opening distance, iris thickness, anterior chamber depth, and pupil size on ultrasound biomicroscopy before and during the Valsalva maneuver. The Valsalva manoeuver was standardized to a pressure of 40 mmHg for 15 s, using a manometer. RESULTS: The mean baseline intraocular pressure changed from 18.86+3.79 to 26.73+4.73 mmHg during Valsalva, (P<0.0001). The anterior chamber angle recess narrowed from 16.62+6.24 to 6.5+4.02 degrees (P<0.0001). There was a significant increase in the thickness of the ciliary body from 0.92+0.25 to 1.17+0.31 mm (P=0.0006) and in the iris thickness from 0.47+0.17 to 0.57+0.11 mm (P=0.007). A significant increase in pupillary diameter (P=0.008) and a decrease in the angle opening distance (P<0.0001) also occurred during Valsalva, whereas there was no significant change in the anterior chamber depth (P=0.056). The angle recess during Valsalva had a positive correlation with the baseline anterior chamber angle (r=0.41, P=0.05) and a negative correlation with the ciliary body thickness (r=-0.52, P=0.046). CONCLUSIONS: The induction of Valsalva maneuver in day-to-day activities can lead to significant anterior segment angle shallowing and can lead to progression from the PAC stage to primary angle closure glaucoma in such predisposed eyes. The presence of a patent laser iridotomy may not prevent irido trabecular apposition during the Valsalva maneuver. PMID- 17417625 TI - Priming, initiation and synchronization of the segmentation clock by deltaD and deltaC. AB - Zebrafish somitogenesis is governed by a segmentation clock that generates oscillations in expression of several Notch pathway genes, including her1, her7 and deltaC. Using a combination of pharmacological inhibition and Mendelian genetics, we show that DeltaD and DeltaC, two Notch ligands, represent functionally distinct signals within the segmentation clock. Using high resolution fluorescent in situ hybridization, the oscillations were divided into phases based on eight distinct subcellular patterns of mRNA localization for 140,000 cells. her1, her7 and deltaC expression was examined in wild-type, deltaD(-/-) and deltaC(-/-) embryos. We identified areas within the tailbud where the clock is set up in the progenitor cells (priming), where the clock starts running (initiation), and where the clocks of neighbouring cells are entrained (synchronization). We find that the clocks of motile cells are primed by deltaD in a progenitor zone in the posterior tailbud and that deltaD is required for cells to initiate oscillations on exiting this zone. Oscillations of adjacent cells are synchronized and amplified by deltaC in the posterior presomitic mesoderm as cell movement subsides and cells maintain stable neighbour relationships. PMID- 17417626 TI - Voltage-dependent anion channels are dispensable for mitochondrial-dependent cell death. AB - Mitochondria are critically involved in necrotic cell death induced by Ca(2+) overload, hypoxia and oxidative damage. The mitochondrial permeability transition (MPT) pore - a protein complex that spans both the outer and inner mitochondrial membranes - is considered the mediator of this event and has been hypothesized to minimally consist of the voltage-dependent anion channel (Vdac) in the outer membrane, the adenine-nucleotide translocase (Ant) in the inner membrane and cyclophilin-D in the matrix. Here, we report the effects of deletion of the three mammalian Vdac genes on mitochondrial-dependent cell death. Mitochondria from Vdac1-, Vdac3-, and Vdac1-Vdac3-null mice exhibited a Ca(2+)- and oxidative stress-induced MPT that was indistinguishable from wild-type mitochondria. Similarly, Ca(2+)- and oxidative-stress-induced MPT and cell death was unaltered, or even exacerbated, in fibroblasts lacking Vdac1, Vdac2, Vdac3, Vdac1-Vdac3 and Vdac1-Vdac2-Vdac3. Wild-type and Vdac-deficient mitochondria and cells also exhibited equivalent cytochrome c release, caspase cleavage and cell death in response to the pro-death Bcl-2 family members Bax and Bid. These results indicate that Vdacs are dispensable for both MPT and Bcl-2 family member-driven cell death. PMID- 17417628 TI - Flies without a spindle checkpoint. AB - Mad2 has a key role in the spindle-assembly checkpoint (SAC) - the mechanism delaying anaphase onset until all chromosomes correctly attach to the spindle. Here, we show that unlike every other reported case of SAC inactivation in metazoans, mad2-null Drosophila are viable and fertile, and their cells almost always divide correctly despite having no SAC and an accelerated 'clock', which is caused by premature degradation of cyclin B. Mitosis in Drosophila does not need the SAC because correct chromosome attachment is achieved very rapidly, before even the cell lacking Mad2 can initiate anaphase. Experimentally reducing spindle-assembly efficiency renders the cells Mad2-dependent. In fact, the robustness of the SAC may generally mask minor mitotic defects of mutations affecting spindle function. The reported lethality of other Drosophila SAC mutations may be explained by their multifunctionality, and thus the 'checkpoint' phenotypes previously ascribed to these mutations should be considered the consequence of eliminating both the checkpoint and a second mitotic function. PMID- 17417627 TI - p53 gain-of-function cancer mutants induce genetic instability by inactivating ATM. AB - Tp53 is the most commonly mutated tumour-suppressor gene in human cancers. In addition to the loss of tumour-suppression function, some missense mutants gain novel oncogenic activities. To elucidate the nature of the gain of function, we introduced the most common p53 cancer mutations (R248W and R273H) independently into the humanized p53 knock-in (HUPKI) allele in mice. Tumour-suppressor functions of p53 are abolished in p53-mutant mice. Several lines of evidence further indicate gain-of-function of p53 mutants in promoting tumorigenesis. p53(R248W) mice rapidly succumb to certain types of cancers not commonly observed in p53(-/-) mice. Interchromosomal translocations, a type of genetic instability rarely observed in p53(-/-) cells, are readily detectable in p53-mutant pre-tumor thymocytes. Although normal in p53(-/-) mouse cells, the G(2)-M checkpoint is impaired in p53-mutant cells after DNA damage. These acquired oncogenic properties of mutant p53 could be explained by the findings that these p53 mutants interact with the nuclease Mre11 and suppress the binding of the Mre11 Rad50-NBS1 (MRN) complex to DNA double-stranded breaks (DSBs), leading to impaired Ataxia-telangiectasia mutated (ATM) activation. Therefore, p53 gain-of function mutants promote tumorigenesis by a novel mechanism involving active disruption of critical DNA damage-response pathways. PMID- 17417629 TI - A functional genomic screen identifies a role for TAO1 kinase in spindle checkpoint signalling. AB - Defects in chromosome-microtubule attachment trigger spindle-checkpoint activation and delay mitotic progression. How microtubule attachment is sensed and integrated into the steps of checkpoint-signal amplification is poorly understood. In a functional genomic screen targeting human kinases and phosphatases, we identified a microtubule affinity-regulating kinase kinase, TAO1 (also known as MARKK) as an important regulator of mitotic progression, required for both chromosome congression and checkpoint-induced anaphase delay. TAO1 interacts with the checkpoint kinase BubR1 and promotes enrichment of the checkpoint protein Mad2 at sites of defective attachment, providing evidence for a regulatory step that precedes the proposed Mad2-Mad1 dependent checkpoint signal amplification step. We propose that the dual functions of TAO1 in regulating microtubule dynamics and checkpoint signalling may help to coordinate the establishment and monitoring of correct congression of chromosomes, thereby protecting genomic stability in human cells. PMID- 17417630 TI - Cdk1 coordinates cell-surface growth with the cell cycle. AB - The mechanisms that control cell growth during the cell cycle are poorly understood. In budding yeast, cyclin dependent kinase 1 (Cdk1) triggers polarization of the actin cytoskeleton and bud emergence in late G1 through activation of the Cdc42 GTPase. However, Cdk1 is not thought to be required for subsequent growth of the bud. Here, we show that Cdk1 has an unexpected role in controlling bud growth after bud emergence. Moreover, we show that G1 cyclin-Cdk1 complexes specifically phosphorylate multiple proteins associated with Cdc24, the guanine nucleotide-exchange factor (GEF) that activates the Cdc42 GTPase. A mutant form of a Cdc24-associated protein that fails to undergo Cdk1-dependent phosphorylation causes defects in bud growth. These results provide a direct link between Cdk1 activity and the control of polarized cell growth. PMID- 17417631 TI - Chemical genetics reveals a complex functional ground state of neural stem cells. AB - The identification of self-renewing and multipotent neural stem cells (NSCs) in the mammalian brain holds promise for the treatment of neurological diseases and has yielded new insight into brain cancer. However, the complete repertoire of signaling pathways that governs the proliferation and self-renewal of NSCs, which we refer to as the 'ground state', remains largely uncharacterized. Although the candidate gene approach has uncovered vital pathways in NSC biology, so far only a few highly studied pathways have been investigated. Based on the intimate relationship between NSC self-renewal and neurosphere proliferation, we undertook a chemical genetic screen for inhibitors of neurosphere proliferation in order to probe the operational circuitry of the NSC. The screen recovered small molecules known to affect neurotransmission pathways previously thought to operate primarily in the mature central nervous system; these compounds also had potent inhibitory effects on cultures enriched for brain cancer stem cells. These results suggest that clinically approved neuromodulators may remodel the mature central nervous system and find application in the treatment of brain cancer. PMID- 17417632 TI - A new function for the fragile X mental retardation protein in regulation of PSD 95 mRNA stability. AB - Fragile X syndrome (FXS) results from the loss of the fragile X mental retardation protein (FMRP), an RNA-binding protein that regulates a variety of cytoplasmic mRNAs. FMRP regulates mRNA translation and may be important in mRNA localization to dendrites. We report a third cytoplasmic regulatory function for FMRP: control of mRNA stability. In mice, we found that FMRP binds, in vivo, the mRNA encoding PSD-95, a key molecule that regulates neuronal synaptic signaling and learning. This interaction occurs through the 3' untranslated region of the PSD-95 (also known as Dlg4) mRNA, increasing message stability. Moreover, stabilization is further increased by mGluR activation. Although we also found that the PSD-95 mRNA is synaptically localized in vivo, localization occurs independently of FMRP. Through our functional analysis of this FMRP target we provide evidence that dysregulation of mRNA stability may contribute to the cognitive impairments in individuals with FXS. PMID- 17417633 TI - CLOCK and NPAS2 have overlapping roles in the suprachiasmatic circadian clock. AB - Heterodimers of CLOCK and BMAL1, bHLH-PAS transcription factors, are believed to be the major transcriptional regulators of the circadian clock mechanism in mammals. However, a recent study shows that CLOCK-deficient mice continue to exhibit robust behavioral and molecular rhythms. Here we report that the transcription factor NPAS2 (MOP4) is able to functionally substitute for CLOCK in the master brain clock in mice to regulate circadian rhythmicity. PMID- 17417634 TI - Choice of cranial window type for in vivo imaging affects dendritic spine turnover in the cortex. AB - Determining the degree of synapse formation and elimination is essential for understanding the structural basis of brain plasticity and pathology. We show that in vivo imaging of dendritic spine dynamics through an open-skull glass window, but not a thinned-skull window, is associated with high spine turnover and substantial glial activation during the first month after surgery. These findings help to explain existing discrepancies in the degree of dendritic spine plasticity observed in the mature cortex. PMID- 17417635 TI - Olfactory processing and behavior downstream from highly selective receptor neurons. AB - In both the vertebrate nose and the insect antenna, most olfactory receptor neurons (ORNs) respond to multiple odors. However, some ORNs respond to just a single odor, or at most to a few highly related odors. It has been hypothesized that narrowly tuned ORNs project to narrowly tuned neurons in the brain, and that these dedicated circuits mediate innate behavioral responses to a particular ligand. Here we have investigated neural activity and behavior downstream from two narrowly tuned ORN types in Drosophila melanogaster. We found that genetically ablating either of these ORN types impairs innate behavioral attraction to their cognate ligand. Neurons in the antennal lobe postsynaptic to one of these ORN types are, like their presynaptic ORNs, narrowly tuned to a pheromone. However, neurons postsynaptic to the second ORN type are broadly tuned. These results demonstrate that some narrowly tuned ORNs project to dedicated central circuits, ensuring a tight connection between stimulus and behavior, whereas others project to central neurons that participate in the ensemble representations of many odors. PMID- 17417636 TI - Comprehensive analysis of the gene encoding filaggrin uncovers prevalent and rare mutations in ichthyosis vulgaris and atopic eczema. AB - We recently reported two common filaggrin (FLG) null mutations that cause ichthyosis vulgaris and predispose to eczema and secondary allergic diseases. We show here that these common European mutations are ancestral variants carried on conserved haplotypes. To facilitate comprehensive analysis of other populations, we report a strategy for full sequencing of this large, highly repetitive gene, and we describe 15 variants, including seven that are prevalent. All the variants are either nonsense or frameshift mutations that, in representative cases, resulted in loss of filaggrin production in the epidermis. In an Irish case control study, the five most common European mutations showed a strong association with moderate-to-severe childhood eczema (chi2 test: P = 2.12 x 10( 51); Fisher's exact test: heterozygote odds ratio (OR) = 7.44 (95% confidence interval (c.i.) = 4.9-11.3), and homozygote OR = 151 (95% c.i. = 20-1,136)). We found three additional rare null mutations in this case series, suggesting that the genetic architecture of filaggrin-related atopic dermatitis consists of both prevalent and rare risk alleles. PMID- 17417637 TI - A QTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase. AB - Grain weight is one of the most important components of grain yield and is controlled by quantitative trait loci (QTLs) derived from natural variations in crops. However, the molecular roles of QTLs in the regulation of grain weight have not been fully elucidated. Here, we report the cloning and characterization of GW2, a new QTL that controls rice grain width and weight. Our data show that GW2 encodes a previously unknown RING-type protein with E3 ubiquitin ligase activity, which is known to function in the degradation by the ubiquitin proteasome pathway. Loss of GW2 function increased cell numbers, resulting in a larger (wider) spikelet hull, and it accelerated the grain milk filling rate, resulting in enhanced grain width, weight and yield. Our results suggest that GW2 negatively regulates cell division by targeting its substrate(s) to proteasomes for regulated proteolysis. The functional characterization of GW2 provides insight into the mechanism of seed development and is a potential tool for improving grain yield in crops. PMID- 17417638 TI - Genetic reconstruction of a functional transcriptional regulatory network. AB - Although global analyses of transcription factor binding provide one view of potential transcriptional regulatory networks, regulation also occurs at levels distinct from transcription factor binding. Here, we use a genetic approach to identify targets of transcription factors in yeast and reconstruct a functional regulatory network. First, we profiled transcriptional responses in S. cerevisiae strains with individual deletions of 263 transcription factors. Then we used directed-weighted graph modeling and regulatory epistasis analysis to identify indirect regulatory relationships between these transcription factors, and from this we reconstructed a functional transcriptional regulatory network. The enrichment of promoter motifs and Gene Ontology annotations provide insight into the biological functions of the transcription factors. PMID- 17417639 TI - Estrogen receptor alpha (ESR1) gene amplification is frequent in breast cancer. AB - Using an Affymetrix 10K SNP array to screen for gene copy number changes in breast cancer, we detected a single-gene amplification of the ESR1 gene, which encodes estrogen receptor alpha, at 6q25. A subsequent tissue microarray analysis of more than 2,000 clinical breast cancer samples showed ESR1 amplification in 20.6% of breast cancers. Ninety-nine percent of tumors with ESR1 amplification showed estrogen receptor protein overexpression, compared with 66.6% cancers without ESR1 amplification (P < 0.0001). In 175 women who had received adjuvant tamoxifen monotherapy, survival was significantly longer for women with cancer with ESR1 amplification than for women with estrogen receptor-expressing cancers without ESR1 amplification (P = 0.023). Notably, we also found ESR1 amplification in benign and precancerous breast diseases, suggesting that ESR1 amplification may be a common mechanism in proliferative breast disease and a very early genetic alteration in a large subset of breast cancers. PMID- 17417640 TI - Production of Ins(1,3,4,5)P4 mediated by the kinase Itpkb inhibits store-operated calcium channels and regulates B cell selection and activation. AB - Antigen receptor-mediated production of inositol-1,4,5-trisphosphate (Ins(1,4,5)P3) in lymphocytes triggers the release of Ca2+ from intracellular stores; this release of Ca2+ results in the opening of store-operated Ca2+ channels in the plasma membrane. Here we report that mice lacking Ins(1,4,5)P3 3 kinase B (Itpkb), which converts Ins(1,4,5)P3 to inositol-1,3,4,5 tetrakisphosphate (Ins(1,3,4,5)P4), had impaired B lymphocyte development and defective immunoglobulin G3 antibody responses to a T lymphocyte-independent antigen. Itpkb-deficient B lymphocytes had the phenotypic and functional features of tolerant B lymphocytes and showed enhanced activity of store-operated Ca2+ channels after B lymphocyte receptor stimulation, which was reversed by the provision of exogenous Ins(1,3,4,5)P4. Our data identify Itpkb and its product Ins(1,3,4,5)P4 as inhibitors of store-operated Ca2+ channels and crucial regulators of B cell selection and activation. PMID- 17417641 TI - Toll-like receptor 9-dependent activation by DNA-containing immune complexes is mediated by HMGB1 and RAGE. AB - Increased concentrations of DNA-containing immune complexes in the serum are associated with systemic autoimmune diseases such as lupus. Stimulation of Toll like receptor 9 (TLR9) by DNA is important in the activation of plasmacytoid dendritic cells and B cells. Here we show that HMGB1, a nuclear DNA-binding protein released from necrotic cells, was an essential component of DNA containing immune complexes that stimulated cytokine production through a TLR9 MyD88 pathway involving the multivalent receptor RAGE. Moreover, binding of HMGB1 to class A CpG oligodeoxynucleotides considerably augmented cytokine production by means of TLR9 and RAGE. Our data demonstrate a mechanism by which HMGB1 and RAGE activate plasmacytoid dendritic cells and B cells in response to DNA and contribute to autoimmune pathogenesis. PMID- 17417642 TI - Solution-phase deposition and nanopatterning of GeSbSe phase-change materials. AB - Chalcogenide films with reversible amorphous-crystalline phase transitions have been commercialized as optically rewritable data-storage media, and intensive effort is now focused on integrating them into electrically addressed non volatile memory devices (phase-change random-access memory or PCRAM). Although optical data storage is accomplished by laser-induced heating of continuous films, electronic memory requires integration of discrete nanoscale phase-change material features with read/write electronics. Currently, phase-change films are most commonly deposited by sputter deposition, and patterned by conventional lithography. Metal chalcogenide films for transistor applications have recently been deposited by a low-temperature, solution-phase route. Here, we extend this methodology to prepare thin films and nanostructures of GeSbSe phase-change materials. We report the ready tuneability of phase-change properties in GeSbSe films through composition variation achieved by combining novel precursors in solution. Rapid, submicrosecond phase switching is observed by laser-pulse annealing. We also demonstrate that prepatterned holes can be filled to fabricate phase-change nanostructures from hundreds down to tens of nanometres in size, offering enhanced flexibility in fabricating PCRAM devices with reduced current requirements. PMID- 17417643 TI - Giant sharp and persistent converse magnetoelectric effects in multiferroic epitaxial heterostructures. AB - Magnetoelectric coupling between magnetic and electrical properties presents valuable degrees of freedom for applications. The two most promising scenarios are magnetic-field sensors that could replace low-temperature superconducting quantum interference devices, and electric-write magnetic-read memory devices that combine the best of ferroelectric and magnetic random-access memory. The former scenario requires magnetically induced continuous and reversible changes in electrical polarization. These are commonly observed, but the coupling constants thus obtained are invalid for data-storage applications, where the more difficult to achieve and rarely studied magnetic response to an electric field is required. Here, we demonstrate electrically induced giant, sharp and persistent magnetic changes (up to 2.3 x 10(-7) s m(-1)) at a single epitaxial interface in ferromagnetic 40 nm La(0.67)Sr(0.33)MnO(3) films on 0.5 mm ferroelectric BaTiO(3) substrates. X-ray diffraction confirms strain coupling via ferroelastic non-180( composite function) BaTiO(3) domains. Our findings are valid over a wide range of temperatures including room temperature, and should inspire further study with single epitaxial interfaces. PMID- 17417644 TI - Extrafluorescent electroluminescence in organic light-emitting devices. AB - Organic light-emitting devices (OLEDs) are a promising technology for flat-panel displays and solid-state lighting. While OLED efficiencies have increased dramatically in recent years, further progress is complicated by the fact that the vast majority of organic materials are fluorescent and therefore emit only from molecular excited states ('excitons') with spin 0, or 'singlet' spin symmetry. Here, we demonstrate the ability to manipulate the fraction of excitons which form as singlets in fluorescent materials by altering the OLED structure. We insert a mixing layer that affects only charge-transfer (CT) states, which are the precursors to excitons. As a result, we triple the singlet fraction and the efficiency of the red fluorophore DCM2. We term fluorescence enhanced by CT spin mixing 'extrafluorescence', and show that its origin is in part an inversion of the usual energetic ordering of the singlet and triplet CT states. PMID- 17417645 TI - A genomic integration method to visualize localization of endogenous mRNAs in living yeast. AB - mRNA localization may be an important determinant for protein localization. We describe a simple PCR-based genomic-tagging strategy (m-TAG) that uses homologous recombination to insert binding sites for the RNA-binding MS2 coat protein (MS2 CP) between the coding region and 3' untranslated region (UTR) of any yeast gene. Upon coexpression of MS2-CP fused with GFP, we demonstrate the localization of endogenous mRNAs (ASH1, SRO7, PEX3 and OXA1) in living yeast (Saccharomyces cerevisiae). PMID- 17417646 TI - Chemoselective probes for metabolite enrichment and profiling. AB - Chemical probes that target classes of proteins based on shared functional properties have emerged as powerful tools for proteomics. The metabolome rivals, if not surpasses, the proteome in terms of size and complexity, suggesting that efforts to profile metabolites would also benefit from targeted technologies. Here we apply the principle of chemoselective probes to the metabolome, creating a general strategy to tag, enrich and profile large classes of small molecules from biological systems. Key to success was incorporation of a protease-cleavage step to release captured metabolites in a format compatible with liquid chromatography-mass spectrometry (LC-MS) analysis. This technology, termed metabolite enrichment by tagging and proteolytic release (METPR), is applicable to small molecules of any physicochemical class, including polar, labile and low mass (<100 Da) compounds. We applied METPR to profile changes in the thiol metabolome of human cancer cells treated with the antioxidant N-acetyl-L cysteine. PMID- 17417647 TI - Multiplexed analysis of glycan variation on native proteins captured by antibody microarrays. AB - Carbohydrate post-translational modifications on proteins are important determinants of protein function in both normal and disease biology. We have developed a method to allow the efficient, multiplexed study of glycans on individual proteins from complex mixtures, using antibody microarray capture of multiple proteins followed by detection with lectins or glycan-binding antibodies. Chemical derivatization of the glycans on the spotted antibodies prevented lectin binding to those glycans. Multiple lectins could be used as detection probes, each targeting different glycan groups, to build up lectin binding profiles of captured proteins. By profiling both protein and glycan variation in multiple samples using parallel sandwich and glycan-detection assays, we found cancer-associated glycan alteration on the proteins MUC1 and CEA in the serum of pancreatic cancer patients. Antibody arrays for glycan detection are highly effective for profiling variation in specific glycans on multiple proteins and should be useful in diverse areas of glycobiology research. PMID- 17417648 TI - Cyclic AMP-regulated exocytosis of Escherichia coli from infected bladder epithelial cells. AB - The superficial bladder epithelium is a powerful barrier to urine and also serves as a regulator of bladder volume, which is achieved by apical exocytosis of specialized fusiform vesicles during distension of the bladder. We report that type 1 fimbriated uropathogenic Escherichia coli (UPEC) circumvents the bladder barrier by harboring in these Rab27b/CD63-positive and cAMP-regulatable fusiform vesicles within bladder epithelial cells (BECs). Incorporation of UPEC into BEC fusiform compartments enabled bacteria to escape elimination during voiding and to re-emerge in the urine as the bladder distended. Notably, treatment of UPEC infected mice with a drug that increases intracellular cAMP and induces exocytosis of fusiform vesicles reduced the number of intracellular E. coli. PMID- 17417649 TI - Noninvasive detection of macrophages using a nanoparticulate contrast agent for computed tomography. AB - Sudden fibrous cap disruption of 'high-risk' atherosclerotic plaques can trigger the formation of an occlusive thrombus in coronary arteries, causing acute coronary syndromes. High-risk atherosclerotic plaques are characterized by their specific cellular and biological content (in particular, a high density of macrophages), rather than by their impact on the vessel lumen. Early identification of high-risk plaques may be useful for preventing ischemic events. One major hurdle in detecting high-risk atherosclerotic plaques in coronary arteries is the lack of an imaging modality that allows for the identification of atherosclerotic plaque composition with high spatial and temporal resolutions. Here we show that macrophages in atherosclerotic plaques of rabbits can be detected with a clinical X-ray computed tomography (CT) scanner after the intravenous injection of a contrast agent formed of iodinated nanoparticles dispersed with surfactant. This contrast agent may become an important adjunct to the clinical evaluation of coronary arteries with CT. PMID- 17417650 TI - Sema3a maintains normal heart rhythm through sympathetic innervation patterning. AB - Sympathetic innervation is critical for effective cardiac function. However, the developmental and regulatory mechanisms determining the density and patterning of cardiac sympathetic innervation remain unclear, as does the role of this innervation in arrhythmogenesis. Here we show that a neural chemorepellent, Sema3a, establishes cardiac sympathetic innervation patterning. Sema3a is abundantly expressed in the trabecular layer in early-stage embryos but is restricted to Purkinje fibers after birth, forming an epicardial-to-endocardial transmural sympathetic innervation patterning. Sema3a(-/-) mice lacked a cardiac sympathetic innervation gradient and exhibited stellate ganglia malformation, which led to marked sinus bradycardia due to sympathetic dysfunction. Cardiac specific overexpression of Sema3a in transgenic mice (SemaTG) was associated with reduced sympathetic innervation and attenuation of the epicardial-to-endocardial innervation gradient. SemaTG mice demonstrated sudden death and susceptibility to ventricular tachycardia, due to catecholamine supersensitivity and prolongation of the action potential duration. We conclude that appropriate cardiac Sema3a expression is needed for sympathetic innervation patterning and is critical for heart rate control. PMID- 17417651 TI - Reverse signaling through GITR ligand enables dexamethasone to activate IDO in allergy. AB - Glucocorticoid-induced tumor necrosis factor receptor (GITR) on T cells and its natural ligand, GITRL, on accessory cells contribute to the control of immune homeostasis. Here we show that reverse signaling through GITRL after engagement by soluble GITR initiates the immunoregulatory pathway of tryptophan catabolism in mouse plasmacytoid dendritic cells, by means of noncanonical NF-kappaB dependent induction of indoleamine 2,3-dioxygenase (IDO). The synthetic glucocorticoid dexamethasone administered in vivo activated IDO through the symmetric induction of GITR in CD4(+) T cells and GITRL in plasmacytoid dendritic cells. The drug exerted IDO-dependent protection in a model of allergic airway inflammation. Modulation of tryptophan catabolism via the GITR-GITRL coreceptor system might represent an effective therapeutic target in immune regulation. Induction of IDO could be an important mechanism underlying the anti-inflammatory action of corticosteroids. PMID- 17417652 TI - Derivation of engraftable skeletal myoblasts from human embryonic stem cells. AB - Human embryonic stem cells (hESCs) are a promising source for cell therapy in degenerative diseases. A key step in establishing the medical potential of hESCs is the development of techniques for the conversion of hESCs into tissue restricted precursors suitable for transplantation. We recently described the derivation of multipotent mesenchymal precursors from hESCs. Nevertheless, our previous study was limited by the requirement for mouse feeders and the lack of in vivo data. Here we report a stroma-free induction system for deriving mesenchymal precursors. Selective culture conditions and fluorescence-activated cell sorting (FACS)-mediated purification yielded multipotent mesenchymal precursors and skeletal myoblasts. Skeletal muscle cells undergo in vitro maturation resulting in myotube formation and spontaneous twitching. We found that hESC-derived skeletal myoblasts were viable after transplantation into the tibialis anterior muscle of SCID/Beige mice, as assessed by bioluminescence imaging. Lack of teratoma formation and evidence of long-term myoblast engraftment suggests considerable potential for future therapeutic applications. PMID- 17417653 TI - Structure of the human GINS complex and its assembly and functional interface in replication initiation. AB - The eukaryotic GINS complex is essential for the establishment of DNA replication forks and replisome progression. We report the crystal structure of the human GINS complex. The heterotetrameric complex adopts a pseudo symmetrical layered structure comprising two heterodimers, creating four subunit-subunit interfaces. The subunit structures of the heterodimers consist of two alternating domains. The C-terminal domains of the Sld5 and Psf1 subunits are connected by linker regions to the core complex, and the C-terminal domain of Sld5 is important for core complex assembly. In contrast, the C-terminal domain of Psf1 does not contribute to the stability of the complex but is crucial for chromatin binding and replication activity. These data suggest that the core complex ensures a stable platform for the C-terminal domain of Psf1 to act as a key interaction interface for other proteins in the replication-initiation process. PMID- 17417654 TI - Bluetongue virus VP4 is an RNA-capping assembly line. AB - Eukaryotic organisms cap the 5' ends of their messenger RNAs by a series of four chemical reactions. Some viruses achieve this using a single molecule; the crystal structure of such an enzyme from bluetongue virus reveals an elongated modular architecture that provides a scaffold for an assemblage of active sites, two contributed by a domain of novel structure. PMID- 17417655 TI - Homology-driven chromatin remodeling by human RAD54. AB - Human RAD51 and RAD54 are key players in homologous recombination, a process that requires homology recognition and strand invasion by a RAD51-single-stranded DNA (ssDNA) nucleoprotein filament and chromatin remodeling by RAD54. Here we use in vitro chromatin reconstitution systems to show that RAD51-ssDNA stimulates RAD54 dependent chromatin remodeling in a homology-dependent, polarity-independent manner. This stimulation was not seen with RAD54B or other remodelers. Chromatin remodeling by RAD54 enabled strand invasion by RAD51-ssDNA on nucleosomal templates, which was homology- and polarity-dependent. Three natural RAD54 mutants found in primary cancer cells showed specific defects in remodeling or in the RAD54-RAD51 interaction. We propose that RAD54 is recruited by RAD51-ssDNA filament to the chromatin of the intact chromosome and that it remodels that chromatin to facilitate accessibility for strand exchange. PMID- 17417656 TI - Iron-regulatory proteins limit hypoxia-inducible factor-2alpha expression in iron deficiency. AB - Hypoxia stimulates erythropoiesis, the major iron-utilization pathway. We report the discovery of a conserved, functional iron-responsive element (IRE) in the 5' untranslated region of the messenger RNA encoding endothelial PAS domain protein 1, EPAS1 (also called hypoxia-inducible factor-2alpha, HIF2alpha). Via this IRE, iron regulatory protein binding controls EPAS1 mRNA translation in response to cellular iron availability. Our results uncover a regulatory link that permits feedback control between iron availability and the expression of a key transcription factor promoting iron utilization. They also show that an IRE that is structurally distinct from, for example, the ferritin mRNA IRE and that has been missed by in silico approaches, can mediate mechanistically similar responses. PMID- 17417657 TI - Severe intravascular haemolysis following minor group mismatched peripheral blood stem cell transplantation. PMID- 17417658 TI - Safety of vaccinating sibling donors with live-attenuated varicella zoster vaccine before hematopoietic stem cell transplantation. AB - Reactivation of varicella zoster virus (VZV), clinically manifested as herpes zoster (HZ) is a common complication after hematopoietic stem cell transplantation (HSCT). The optimum prophylaxis for this disease has not been defined. In this study, we examined the effects of vaccinating donors with a live attenuated vaccine with particular reference to their immune responses and the outcome of HSCT patients. Forty prospective HLA-matched sibling donors were vaccinated before HSCT. There were humoral immune responses in both sero-positive (P<0.01) and sero-negative (P=0.058) donors. Cellular immune response was assayed in 26 donors. Significant correlation was observed between cellular immune responses as enumerated by thymidine incorporation and interferon gamma secretion (P<0.001) and the latter was used in subsequent analyses. Significant response was observed in sero-negative (6/26) and a group of sero-positive (13/26) donors while 7/26 sero-positive donors showed no response. Thirty-four HSCT were performed. These patients have a lower, albeit insignificant, risk of HZ compared with historical controls and only 3/34 patients developed single dermatomal HZ at 6, 9 and 28 months after HSCT. No patients developed VZV-related mortality. Vaccinating donors with live-attenuated VZV vaccine was safe, but whether it confers a significant protection to the patients would require further study. PMID- 17417659 TI - The potential role of recombinant activated FVII in the management of critical hemato-oncological bleeding: a systematic review. AB - Recombinant activated factor VII (rFVIIa) is an hemostatic agent that was originally developed for the treatment of hemorrhage in patients with hemophilia and inhibitors. However, in the last few years rFVIIa has been employed with success in a broad spectrum of congenital and acquired bleeding conditions. In this systematic review we present the current knowledge on the use of this drug in patients suffering from hemato-oncological disorders, which are quite commonly complicated by severe hemorrhage. On the whole, data in the literature suggest a potential role for rFVIIa in the management of bleeding unresponsive to standard therapy in patients with hematological malignancies, including those undergoing bone marrow transplant. However, the vast majority of the currently available data are derived from uncontrolled studies including single cases or small series of patients. Thus, further trials with larger numbers of patients are needed to establish the most appropriate doses and timing of rFVIIa and to assess its efficacy and safety in this setting. PMID- 17417660 TI - Early recovery of aggressive cytotoxic cells and improved immune resurgence with post-transplant immunotherapy for multiple myeloma. AB - A phase I/II trial evaluated early administration and dose escalation of interleukin (IL)-2 with granulocyte macrophage colony stimulating factor (GM-CSF) post-transplant. Following melphalan (200 mg/m(2)) and an autologous transplant, IL-2 was initiated (day 0) and continued for 4 weeks. GM-CSF (250 mcg/m(2)/day) began on day 5. Fifteen of 19 patients completed therapy. No treatment-related deaths occurred. IL-2 (1 x 10(6) IU/m(2)/day) was not tolerated in two of six patients due to > or =grade 3 fatigue/diarrhea (n=1) or supraventricular tachycardia (n=1). The maximum tolerated dose of IL-2 was 6 x 10(5) IU/m(2)/day; this dose was well tolerated by 11 of 13 patients. Neutrophil and platelet engraftment occurred on day 13 (median; range 10-17 days) and day 13 (median; range 0-74 days), respectively. When compared to control patients, there was a marked increase in the number of CD3+ T cells (P=0.005), CD4+ T cells (P=0.01), CD8+ T cells (P=0.001) and CD4+CD25+Treg cells (P=0.015) post-transplant. Cytotoxicity directed against myeloma cells was markedly increased when compared to control patients (P=0.017). This unique trial design using early administration of IL-2 with GM-CSF during the period of lymphodepletion, demonstrated a marked increase in the number and function of early cytotoxic effector T cells, without suppression of engraftment. PMID- 17417661 TI - Cognitive function in the acute course of allogeneic hematopoietic stem cell transplantation for hematological malignancies. AB - The aim of the study was to assess cognitive performance in patients with hematological malignancies before, and 3 months after, allogeneic hematopoietic stem cell transplant (HSCT). A consecutive sample of 39 patients was assessed before admission with a comprehensive neuropsychological test battery and health related quality-of-life (HRQoL) questionnaires; 19 of these patients were retested around 100 days post HSCT. Test results were compared with normative data and revealed minimal differences at both time points in the level of group means. One parameter - simple reaction time - was significantly worse (prolonged) at second measurement after HSCT. According to the definition of an impairment score (more than three impaired functions), 26% of patients were classified as impaired before as well as after HSCT. Neuropsychological test results did not vary systematically according to medical variables such as extent of pretreatment, graft-versus-host-disease (GvHD) and kind of conditioning protocol. As a dimension of HRQoL, self-rated cognitive function was in the normal range before and after HSCT. Significant correlations between HRQoL and neuropsychological parameters were related to symptom scales. This study showed impairments of neuropsychological performance for a subgroup of patients before and after allogeneic HSCT. Systematic effects of conditioning, medical variables or self-rated HRQoL could not be observed. PMID- 17417662 TI - New clinical grading system for chronic GVHD predicts duration of systemic immunosuppressive treatment and GVHD-specific and overall survival. AB - We investigated outcomes according to a new clinical grading system for chronic graft-versus-host disease (chronic GVHD) in 38 patients who developed chronic GVHD after an allogeneic hematopoietic stem cell transplantation. We categorized the patients into three grade groups, namely, grade I, grade II and grade III, according to the presence of three risk factors: extensive skin involvement, thrombocytopenia (TP) and progressive type of onset. Sixteen patients were classified into grade 1, 19 into grade II and three into grade III. The probability of withdrawal of systemic immunosuppression (IST) at 1, 2 and 3 years was 61, 76 and 87%, respectively. Patients with grades 2 or 3 chronic GVHD had prolonged duration of systemic IST compared to grade 1 (P=0.043). The probability of GVHD-specific survival (GSS) at 5 years was 52%. Twenty-two of 38 patients with chronic GVHD were still alive and the estimated 3-year overall survival (OS) rate was 60%, whereas that for the group with chronic GVHD grade I and grade II+III was 64 and 48% (P<0.05). Multivariate analysis showed that prior occurrence of acute GVHD, chronic GVHD grade, serum bilirubin over 1.5 mg/dl, date of diagnosis of chronic GVHD (day 150) and transplantation risk factor were independent prognostic factors for GSS and OS. PMID- 17417663 TI - Correction of immunodeficiency associated with NEMO mutation by umbilical cord blood transplantation using a reduced-intensity conditioning regimen. PMID- 17417664 TI - Reconstitution of adenovirus-specific cell-mediated immunity in pediatric patients after hematopoietic stem cell transplantation. AB - Adenovirus (adv) is a significant cause of morbidity and mortality in pediatric hematopoietic stem cell transplant recipients, and control of infection seems to require antigen-specific T cells. We evaluated the recovery of adv-specific cellular immunity in this patient population related to degree of T-cell immunosuppressive therapy and compared this to adv cellular immunity of normal donors. Over 12 months, we monitored for adv DNA in stool and blood of patients and in the blood of a normal donor group. Twenty-two pediatric hematopoietic stem cell transplant (HSCT) patients (14 months-20 years) who received matched-related (MRD n=6), mismatched related (Haplo n=6) or matched unrelated donor (MUD n=10) grafts, were followed and results compared to healthy controls (n=8). Adv was detected by polymerase chain reaction in blood and/or stool from 81.8% of patients on at least one occasion post-HSCT, but only 68% of patients developed symptomatic adv infections. Recovery of adv-specific T cells was significantly delayed in the MUD and Haplo recipients, whereas recovery in the MRD group was similar to levels detected in healthy donors within 30 days post-transplant. In conclusion, recipients of alternative donor transplants at our institution have significantly delayed adv-specific cellular immune recovery, which correlates to an increased risk of adv-associated morbidity and mortality. PMID- 17417665 TI - Interaction of Toll-like receptors with bacterial components induces expression of CDX2 and MUC2 in rat biliary epithelium in vivo and in culture. AB - The mechanism of transformation of biliary epithelium leading to intestinal metaplasia, which is significantly involved in biliary diseases, remains unclear. CDX2, an intestine-specific transcription factor, is thought to regulate intestinal mucin MUC2 (mucus core protein) expression. We took advantage of polycystic kidney (PCK) rats as a model of chronic suppurative cholangitis with intestinal metaplasia and of cultured biliary epithelial cells (BECs) from PCK rats to clarify the causal relation between bacterial components such as pathogen associated molecular patterns (PAMPs) and the development of intestinal metaplasia of bile ducts. Histological, immunohistochemical, and in situ hybridization studies were conducted in PCK rat livers. In cultured BECs, CDX2 and MUC2 were expressed following treatment with PAMPs and inhibitors (anti-Toll like receptor (TLR)2/TLR4 antibody, nuclear factor-kappaB (NF-kappaB) inhibitor MG132). Chronic suppurative cholangitis with intestinal metaplasia developed as the PCK rats aged, and intestinal metaplasia and aberrant CDX2 and MUC2 expression developed in parallel. Intraluminal bacteria and the expression of TLR2 and TLR4 in BECs were demonstrated in the bile ducts, showing chronic suppurative cholangitis. In cultured BECs, treatment with PAMPs induced upregulation of CDX2 and MUC2 expression, and this effect was abolished by pretreatment with anti-TLR2 and anti-TLR4 antibody and MG132. A knockdown of CDX2 by CDX2 small interfering RNA inhibited MUC2 expression in cultured BECs induced by PAMPs, and transfection of CDX2 expression vector induced MUC2 expression. In conclusion, bacterial components may induce upregulation of the CDX2 expression followed by MUC2 expression via TLR and the NF-kappaB system in cultured BECs, and could be related to the development of intestinal metaplasia of the bile ducts. PMID- 17417666 TI - Predicting impact factor one year in advance. AB - The first impact factor (IF) to reflect the sole efforts of a new editorial team occurs 4 years into what is usually a 5-year editorship, owing to the lag times of: paper accrual and publication, accumulation of citations in derivative literature, and compiling of such citations by the Thomson ISI Web of Knowledge service. Through weekly collection of citation data from the Web of Science over the past 2 years, we now demonstrate that the evolution of IF can be tracked weekly over the course of a calendar year, enabling prediction of the next year's IF beginning at the middle of the previous year. The methodology used to track the developing IF for Lab Invest is presented in this study and a prediction made for the 2006 IF, along with IF predictions for other general pathology journals (American Journal of Pathology, Journal of Pathology, Modern Pathology, American Journal of Surgical Pathology, and Human Pathology). Despite the fact that the 2006 IF for Lab Invest will not be issued until June 2007, it became apparent as early as July 2006 that the Lab Invest IF would be greatly improved over 2004 and 2005 by a predicted 0.5 units. However, as important as IF can be to a journal, it is vital not to let IF considerations influence every aspect of the editors' decisions. Rather, the significance of early prediction lies in earlier validation of editorial policies for journal management as a whole, and reassurance that the philosophy for journal operations is on track. PMID- 17417667 TI - Changes of gene expression of iron regulatory proteins during turpentine oil induced acute-phase response in the rat. AB - In the present study, turpentine oil was injected in the hind limb muscle of the rat to stimulate an acute-phase response (APR). The changes in the gene expression of cytokines and proteins known to be involved in the iron regulatory pathway were then studied in the liver and in extra-hepatic tissue. In addition to the strong upregulation of interleukin-6 (IL-6) and IL-1 beta observed in the inflamed muscle, an upregulation of the genes for IL1-beta and tumor necrosis factor-alpha, but not IL-6, were detectable in the liver. Hepatic Hepc gene expression increased to a maximum at 6 h after the onset of APR. An upregulation of transferrin, transferrin receptor 1 (TfR1), TfR2, ferritin-H, iron responsive element binding protein-1 (IRP1), IRP2 and divalent metal transporter gene expression was also found. Hemojuvelin (Hjv)-, ferroportin 1-, Dcytb-, hemochromatosis-gene- and hephaestin gene expression was downregulated. Hepcidin (Hepc) gene expression was not only detectable in extra-hepatic tissues such as heart, small intestine, colon, spleen and kidney but it was also upregulated under acute-phase conditions, with the Hjv gene being regulated antagonistically. Fpn-1 gene expression was downregulated significantly in heart, colon and spleen. Most of the genes of the known proteins involved in iron metabolism are expressed not only in the liver but also in extra-hepatic tissues. Under acute-phase conditions, acute-phase cytokines (eg IL-6) may modulate the gene expression of such proteins not only in the liver but also in other organs. PMID- 17417668 TI - Macrophage Serum-Based Adhesion to Plasma-Processed Surface Chemistry is Distinct from That Exhibited by Fibroblasts. AB - Plasma-polymerized films deposited from AlAm, HxAm, NVP, NVFA, AA and FC were compared to TCPS and PS surfaces in supporting cellular attachment, viability, and proliferation in serum-based culture in vitro for extended periods of time (>7 d). Surface patterns were created using multi-step depositions with physical masks. Cell adhesion in the presence of serum was compared for (monocyte-) macrophage and fibroblast cell lines. Cellular response was tracked over time, reporting adhesive behavior, proliferative rates, and morphological changes as a function of surface chemistry. Micropatterned surfaces containing different surface chemistries and functional groups (e.g. -NH(2), -COOH, -CF(3)) produced differential cell adhesive patterns for NIH 3T3 fibroblasts compared to J774A.1, RAW 264.7 or IC-21 (monocyte-) macrophage cell types. Significantly, macrophage adhesion is substantial on surfaces where fibroblasts do not adhere under identical culture conditions. PMID- 17417669 TI - Parenting Interventions for Drug-Dependent Mothers and Their Young Children: The Case for an Attachment-Based Approach. AB - Maternal substance abuse is the most common factor involved when children come to the attention of the child welfare system. Although there is a clear need for clinical trials to evaluate parenting interventions for drug-dependent women, few studies to date have systematically examined the efficacy of interventions for this population. We first review six published reports of outpatient interventions that aimed to enhance the caregiving skills of substance-abusing mothers caring for children between birth and 5 years of age. After discussing implications of these preliminary studies, we then describe an attachment-based intervention that addresses these implications and has demonstrated preliminary feasibility in a pilot trial. PMID- 17417670 TI - Novel synthon for incorporating 1,3-dimethyl imidazolium group into molecular architecture. AB - The synthesized 1,3-dimethylated imidazolium carbaldehydes serves as synthons for incorporating a permanently cationic imidazolium group into molecular framework. The utility of new synthon was demonstrated in a variety of reactions: Knoevenagel, Wittig, Schiff base formation, based mediated electrophilic substitution and oxidation, including synthesis of the natural product norzooanemonin. PMID- 17417671 TI - [Theory and practice of primary cancer prevention]. AB - The primary aim of cancer prevention is to stop carcinogens from entering the body. Since the low doses involved in carcinogenesis do not cause true toxicological effects, usual toxicological analytic methods do not allow the detection of the early effects of carcinogens. Exposure to chemical carcinogens causes damage to nuclear chromatin, the most vulnerable part of the cell, by inducing DNA damage, chromosomal abnormalities and mutations, which foreshadow the danger of cancer development. In such cases intervention is possible in two ways. On the one hand, we attempt to remove the causative agent from the environment, while on the other we aid the elimination of somatic mutations. The latter is called active prevention; the introduction of substances into the body that can help the elimination of defective cells (apoptosis induction) or stop processes responsible for elongation errors (i.e. with antioxidants). Concerning our own studies, we present the results of 25 years of research on the genotoxicological characteristics of workers exposed to various chemicals, which show that active prevention can in fact be effective in conjunction with information on specific biomarkers. We present in detail the genotoxic changes found in hospital nurses who routinely administer intravenous cytostatic therapy, and the relationship of these changes to their immunotoxic and clinical laboratory parameters. Genotoxic substances decrease the oxidative burst and natural killer (NK) cell activity, which may explain the immunosuppressive effects of occupational exposures. We also present the detailed results of a follow-up study involving two groups of industrial workers. We monitored the status of workers involved in benzene production for 15 years and of asphalt industry workers for 8 years. In both studies we concluded that genotoxic effects can be decreased by ensuring appropriate working conditions, while a temporary lapse in these conditions or accidental changes lead to increases in genotoxic parameters. Since genotoxic effects develop over an extended period (4-5 months), they are independent of hygienic conditions at any single inspection and, thus, their detection also offers a way to ascertain true exposure levels. Our studies also show a connection between genotoxic effects and immune function, which is adversely affected not only by occupational exposures, but also by medications and smoking. From our results with workers in the oil and asphalt industries, we concluded that the levels of chromosomal aberrations (CAs) and sister chromatid exchange (SCE) increase in proportion to exposure levels and decrease with a certain delay following the attenuation of the exposure. We could not detect an increased frequency of any chronic disease in industrial workers. The increased numbers of iron deficiency anemia and thyroid disease in nurses providing cytostatic therapy was, however, related to their occupational exposure. PMID- 17417672 TI - [Quality control of HER2 immunohistochemistry--results from a Hungarian study]. AB - Whether a breast carcinoma is HER2-positive or -negative has a significant impact on its treatment. The most common method of evaluating the HER2 status of breast tumors is immunohistochemistry. Preliminary data suggest that the proportion of HER2-positive tumors in Hungary shows a wide range from laboratory to laboratory, and overall it just reaches the bottom levels of the incidence reported in the literature (12% 3+ and 11% 2+ cases). A 3-round quality-control test was implemented on a voluntary and anonymous basis. Participating laboratories had to immunostain and evaluate 4 tumor samples per circulation, according to their daily routine. The authors of the present article gave an expert opinion in all cases, and this was compared with the individual laboratories' evaluation and the real FISH-controlled HER2 status of the samples. On the basis of the participants' and experts' evaluation 22/218 and 21/218 HER2 3+ cases were underscored, corresponding to an underevaluation rate of 10%. As most samples were from HER2-positive tumors, overscoring was less common (1%), but reached 5% (individual laboratories' evaluation) or 7% (expert evaluation) when the 2+ scoring of negative cases was also considered. Each case was discussed interactively with the participants and technical advise was also given when deemed necessary. The evaluation of the HER2 status of breast cancers gives reliable results only if adequate quality control measures are implemented, and this study was an important step in this respect. PMID- 17417673 TI - [Dosimetric evaluation of interstitial high-dose-rate implants for localised prostate cancer]. AB - PURPOSE: Quantitative evaluation of dose distributions of high-dose-rate prostate implants in order to make a later comparison with clinical outcome. MATERIAL AND METHODS: Treatment plans of 169 implants for 161 patients were evaluated using dose-volume histograms. The planning was based on transrectal US imaging and 10 Gy (100%) dose was prescribed to the surface of the prostate. The tolerance dose to urethra and rectum was 125% and 80%, respectively. The volume of the prostate was measured, and its fraction receiving 90%, 100%, 150% and 200% of the prescribed dose was calculated (V90, V100, V150, V200). The dose delivered to 90% of the prostate volume (D90) and the minimum dose in the prostate (Dmin) were determined. The dose nonuniformity ratio (DNR) and the dose homogeneity index (DHI) were calculated to quantify the dose homogeneity. The coverage index (CI) was determined, and the dose conformality to the target volume was assessed with the use of the conformal index (COIN). Maximal dose to rectum (Dr) and urethra (Du) reference points, dose to volume of 2 cm3 of the rectum (D2) and 0.1 cm3 and 1% of the urethra (D0.1, D1) were determined, too. Correlation analysis was performed between point and volume doses. In most patients in-vivo dose measurement was performed in the rectum with semiconductor detectors. RESULTS: The median number of needles was 16, the mean prostate volume was 25.5 cm3. The mean V90, V100, V150 and V200 were 98%, 94%, 41% and 14%, respectively. The mean D90 was 107%, and the Dmin was 82%. The mean dose to rectum and urethra reference points was 75% and 120%, respectively. The mean volume doses were D2=49% for the rectum, D0.1=128% and D1=143% for the urethra. The correlation coefficients were: R(Dr,D2)=0.69, R(Du,D0.1)=0.55, R(Du,D1)=0.23. The mean DNR was 0.39, while the DHI was 0.57. On average, 94% of the target volume received at least the prescribed dose (CI=0.94) and the mean COIN was 0.64. The mean maximal measured dose in the rectum was 2.67 Gy. CONCLUSIONS: Our US-based treatment plans based on the real positions of catheters provided acceptable dose distributions. In the majority of our cases the dose to urethra and rectum was kept below the defined tolerance level. The dose of rectal reference points correlated well with rectal dose-volume parameters but for urethra dose determination the use of the D1 volumetric parameter is recommended. Finding correlations between dose-volume parameters and clinical side effects requires further analysis. PMID- 17417674 TI - [Histopathological prognostic factors of malignant melanoma]. AB - The incidence of melanocytic skin lesions, including malignant melanoma has increased in the past few years; histopathologists and dermatopathologists have to face them more often. The correct treatment of melanoma patients by the oncodermatologist and oncologist is based on the histopathological report containing the most important histological prognostic factors. However, the accurate interpretation of these factors may be difficult in the everyday practice, especially in reporting tumor thickness, the level of invasion, the type of exulceration and regression. It is important to standardize the content of the histopathological reports in a reproducible way. PMID- 17417675 TI - [The role of emotional labour in oncology]. AB - Oncologists and related health care professionals (HCPs) do not only have to follow professional protocols in their everyday work, but also have to communicate proper attitudes towards patients suffering from malignant diseases. This task is often a heavier load than the implementation of professional activities themselves. The present article is based on a survey on HCP work motivation, employment parameters and correlations with emotional labour. Fifty oncology HCPs at Debrecen University Medical Health Sciences Centre volunteered to participate in this survey containing 20 simple-choice questions. More than 90 percent of HCPs make an effort to hide their emotional state, giving way to possible negative side effects. The survey showed significant differences between the level of emotional labour of those working in the field of oncology longer or shorter than ten years. Surface and deep emotional labour is more frequent among professionals already working in oncology for a longer period of time. This can serve us with explanation to the burn-out syndrome so frequent in this profession. To diminish the load of emotional labour, healthcare institutes have to aim at hiring employees that spontaneously fit the emotional and behavioural norms facing them, and do not need officially prescribed behavioural norms for everyday work. Their constant need for respect and appreciation of their values must be kept in mind, because the capability of genuine emotional labour diminishes parallel to the number of years spent in work. PMID- 17417676 TI - [Human recombinant erythropoietin-alpha increases the efficacy of irradiation in preclinical model]. AB - According to recent data erythropoietin receptor (EPOR) is expressed not only by bone marrow erythroid progenitors but by endothelial- and cancer cells and it was suggested that erythropoietin (EPO) may affect their functions as well. We have analyzed the effects of recombinant human erythropoietin-alpha (rHuEPOalpha) on radiation sensitivity of EPOR+ human epidermoid carcinoma (A431) xenograft model. In vivo rHuEPOalpha treatment was started after tumor cell inoculation into SCID mice. 5 Gy irradiation was performed on day 14, the effect of which was measured on day 22. Hemoglobin level, tumor-associated microvessels and HIF-1alpha expression of the xenograft were monitored during the experiment. rHuEPOalpha administration prevented the development of tumor-induced anemia of SCID mice and reduced the level of HIF-1alpha expression of the xenograft tumor without affecting tumor growth. We have found that rHuEPOalpha treatment significantly increased the efficacy of antitumor effect of irradiation which was partly mediated by complex effects on tumor-associated microvessels. In vitro rHuEPOalpha did not affect proliferation of A431 cells but enhanced the antiproliferative and proapoptotic effects of irradiation. We concluded that rHuEPOalpha administration positively modulated the antitumoral effects of irradiation at least by two pathways, decreasing systemic hypoxia resulting in decreased tumoral HIF-1alpha expression and enhancing direct effects on tumor associated microvessels. PMID- 17417677 TI - [Retroperitoneal adenoma]. AB - The authors describe a tumor localized next to the left adrenal gland. On histological examination the lipid-rich tumor was found to be partly a solid adrenocortical adenoma and partly a tubular oncocytoma. PMID- 17417678 TI - Tetracycline-inducible expression systems: new strategies and practices in the transgenic mouse modeling. AB - To accurately analyze the function of transgene(s) of interest in transgenic mice, and to generate credible transgenic animal models for multifarious human diseases to precisely mimic human disease states, it is critical to tightly regulate gene expression in the animals in a conditional manner. The ability to turn gene expression on or off in the restricted cells or tissues at specific time permits unprecedented flexibility in dissecting gene functions in health and disease. Pioneering studies in conditional transgene expression have brought about the development of a wide variety of controlled gene expression systems, which meet this criterion. Among them, the tetracycline-controlled expression systems (e.g. Tet-off system and Tet-on system) have been used extensively in vitro and in vivo. In recent years, some strategies derived from tetracycline inducible system alone, as well as the combined use of Tet-based systems and Cre/lox P switching gene expression system, have been newly developed to allow more flexibility for exploring gene functions in health and disease, and produce credible transgenic animal models for various human diseases. In this review these newly developed strategies are discussed. PMID- 17417679 TI - Identification of nuclear factor-kappaB responsive element within the neuronal nitric oxide synthase exon 1f-specific promoter. AB - Transcriptional regulation of the neuronal nitric oxide synthase gene (nNOS) is particularly complex as 12 distinct transcripts derived from different first exons are expressed in a tissue- and cell-specific manner. The exon 1f mRNA is relatively highly expressed in nervous system and relies upon exon 1f-specific promoter activity. Using conventional and real-time reverse transcription polymerase chain reaction, we found exon 1f mRNA was the major transcript of the nNOS gene in human neuroblastoma SK-N-SH cells. We analyzed a 1090 bp fragment of 1f promoter by TRANSFAC-TESS and Match softwares and luciferase assay, and found an important positive transcriptional regulation region that contained a putative nuclear factor (NF)-kappaB binding site. Subsequently, using electrophoresis mobility shift and chromatin immunoprecipitation assays, we identified this site to be the NF-kappaB responsive element, a crucial positive regulator in the activation of the nNOS 1f promoter. Taken together, our study identified an NF kappaB responsive element within nNOS 1f promoter and showed that it plays an important role in the transactivation of nNOS 1f mRNA, the major transcript of nNOS in SK-N-SH cells. PMID- 17417680 TI - Characterization, evolution and expression of the calmodulin1 genes from the amphioxus Branchiostoma belcheri tsingtauense. AB - Two full-length cDNAs, named CaM1a and CaM1b, encoding the highly conserved calmodulin1 (CaM1) proteins, were isolated from the cDNA library of amphioxus Branchiostoma belcheri tsingtauense. There are only two nucleotide differences between them, producing one amino acid difference between CaM1a and CaM1b. Comparison of the amino acid sequence of CaM1 reveals that the B. belcheri tsingtauense CaM1a is identical with CaM1 proteins of B. floridae and B. lanceolatum, Drosophila melanogaster CaM, ascidian Halocynthia roretzi CaMA and mollusk Aplysia californica CaM, and CaM1b differs only at one position (138, Asn to Asp). The phylogenetic analysis indicates that the CaM1 in all three amphioxus species appears to encode the conventional CaM and CaM2 might be derived from gene duplication of CaM1. Southern blot suggests that there are two copies of CaM1 in the genome of B. belcheri tsingtauense. Northern blot and in situ hybridization analysis shows the presence of two CaM1 mRNA transcripts with various expression levels in different adult tissues and embryonic stages in amphioxus B. belcheri tsingtauense. The evolution and diversity of metazoan CaM mRNA transcripts are also discussed. PMID- 17417681 TI - Polymorphism analyses of hepatitis B virus X gene in hepatocellular carcinoma patients from southern China. AB - Chronic hepatitis B virus (HBV) infection is one of the major causes of hepatocellular carcinoma (HCC), and the HBV X (HBx) gene plays a critical role in the molecular pathogenesis of HBV-related HCC. We have investigated whether there are particular HBx gene mutations associated with HCC in patients from southern China. The HBx gene was examined in 51 paraffin-embedded tumor tissue samples from patients with HCC and 25 serum samples from the HBV carrier by nested polymerase chain reaction (PCR), single-stranded conformational polymorphism and heteroduplex analysis. The HBx genes with potentially important mutations from tumor tissue samples were cloned, sequenced and aligned with the published HBx gene sequence. HBV genotypes in tumor tissue samples were analyzed by nested PCR. Analyses of HBx gene polymorphism showed that 31.3% of HBx gene fragments in tumor tissue samples had a special pattern. A common deletion at nt 382-400 of the HBx gene accompanied by 29 point mutations was detected in four randomly selected tumor tissue samples with this pattern which caused a frame-shift in the HBx open reading frame with a new stop codon at nt 1818, resulting in an HBx polypeptide chain truncated at the C end in these cases. Among the four randomly selected samples, three were HBV genotype B, and one was not detected by our present assay. In another tumor tissue sample, amplification of the full-length HBx gene yielded a shorter fragment. Sequencing of this fragment revealed a 264 bp deletion between nt 1577 and 1840 of the HBV gene. These results suggest that HBx gene mutation occurs frequently in HCC samples, and the deletion at nt 382 400 of the HBx gene might play a role in carcinogenesis of HCC in southern China. PMID- 17417682 TI - Inhibition of monosodium urate monohydrate-mediated hemolysis by vitamin E. AB - Microcrystals of monosodium urate monohydrate (MSUM) induce cytolysis and hemolysis in erythrocytes. In this report, we studied the effect of vitamin E on MSUM-mediated hemolysis in human erythrocytes. Vitamin E significantly inhibited hemolysis induced by MSUM. The hydroxyl group in the chromanol ring of vitamin E is dispensable for protecting erythrocytes against hemolysis induced by MSUM, indicating that the inhibitory effect of vitamin E is not due to its antioxidant properties. However, both the chromanol ring and the isoprenoid side chain are important for vitamin E to suppress MSUM-induced hemolysis. Our current study suggests that vitamin E inhibits hemolysis induced by MSUM as a membrane stabilizer. PMID- 17417683 TI - Inhibition of tumor growth in mice by endostatin derived from abdominal transplanted encapsulated cells. AB - Endostatin, a C-terminal fragment of collagen 18a, inhibits the growth of established tumors and metastases in vivo by inhibiting angiogenesis. However, the purification procedures required for large-scale production and the attendant cost of these processes, together with the low effectiveness in clinical tests, suggest that alternative delivery methods might be required for efficient therapeutic use of endostatin. In the present study, we transfected Chinese hamster ovary (CHO) cells with a human endostatin gene expression vector and encapsulated the CHO cells in alginate-poly-L-lysine microcapsules. The release of biologically active endostatin was confirmed using the chicken chorioallantoic membrane assay. The encapsulated endostatin-expressing CHO cells can inhibit the growth of primary tumors in a subcutaneous B16 tumor model when injected into the abdominal cavity of mouse. These results widen the clinical application of the microencapsulated cell endostatin delivery system in cancer treatment. PMID- 17417684 TI - Organic solvents mediate self-assembly of GAV-9 peptide on mica surface. AB - Self-assembly of peptides into fibrils and other morphologies has attracted much attention in many fields, especially in nanofabrication, pathology and biochemistry. In this paper, self-assembly of GAV-9 peptide in organic solvents, ethanol and acetone, was investigated using atomic force microscopy (AFM) and attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR). The results indicated that GAV-9 self-assembled into various nanostructures in both solvents after deposited and evaporated on mica. Fibrils with beta-sheet conformation were observed in both solvents when the peptide concentration was higher than 280 microM. However, ordered fibrils with beta-sheet conformation were formed in ethanol, but not in acetone, with a peptide concentration ranging from 7 microM to 28 microM. We attribute the formation of various nanostructures to the different physicochemical properties of the polar organic solvents on the self-assembly of GAV-9 peptide. PMID- 17417685 TI - Immune responses and protective efficacy induced by 85B antigen and early secreted antigenic target-6 kDa antigen fusion protein secreted by recombinant bacille Calmette-Guerin. AB - In an attempt to improve immune responses and protective efficacy, we constructed two recombinant bacille Calmette-Guerin (rBCG) strains expressing an 85B antigen (Ag85B) and early secreted antigenic target-6 kDa antigen (ESAT6) of Mycobacterium tuberculosis (MTB) fusion protein. Both rBCG strains have the same protein insertion but in a different order (Ag85B-ESAT6 and ESAT6-Ag85B). The cultured supernatant of rBCG strains and the sera from the mice immunized with the fusion protein Ag85B-ESAT6 or ESAT6-Ag85B formed a band with a fraction size of 37 kDa, equalivalent to the sum of Ag85B and ESAT6. Six weeks after BALB/c mice were immunized with BCG or rBCG, spleen lymphocytes showed significant proliferation in response to culture filtrate protein of MTB. Compared with the BCG group, mice vaccinated with rBCG elicited a high level increase of immunoglobulin G antibodies to culture filtrate protein in the serum. The gamma interferon levels in the lymphocyte culture medium supernatants increased remarkably in the rBCG1 group, significantly higher than that of the BCG immunized group (p<0.05). Four weeks after vaccination, mice were infected with M. tuberculosis H37Rv and a dramatic reduction in the numbers of MTB colony forming units in the spleens and lungs was observed in the two rBCG immunization groups. Although these rBCG strains were more immunogenic, their protective effect was comparable to the classical BCG strain, and there were no significant differences between two rBCG groups (p>0.05). PMID- 17417686 TI - Cell-specific regulation of APOBEC3F by interferons. AB - Human cytidine deaminase APOBEC3F (A3F) has broad anti-viral activity against hepatitis B virus and retroviruses including human immunodeficiency virus type 1. However, its regulation in viral natural target cells such CD4+ T lymphocytes, macrophages, and primary liver cells has not been well studied. Here we showed that A3F was up-regulated by interferon (IFN)-alpha in primary hepatocytes and multiple liver cell lines as well as macrophages. Although the IFN-alpha signaling pathway was active in T lymphoid cells and induction of other IFN stimulated genes such as PKR was detected, A3F and APOBEC3G (A3G) were not induced by IFN-alpha in these cells. Thus, additional factors other than known IFN-stimulated genes also regulated IFN-alpha-induced A3F expression distinctly. A3F and A3G expression levels in primary hepatocytes, especially after IFN-alpha stimulation, were comparable to those in CD4+ T lymphocytes in some individuals. Significant variations of A3F and A3G expression in primary hepatocytes from various subjects were observed. Individual variations in A3F and/or A3G regulation and expression might influence the clinical outcomes of hepatitis B infection. PMID- 17417687 TI - Different transcription profiles of SOCS-3, ob and IGF-I genes and their possible correlations in obese and lean pigs. AB - Pig breeds have significant differences in fat deposition and muscle development ability. However, the molecular mechanism behind these differences is still unknown. In this study, the expression patterns of three candidate genes, suppressor of cytokine signaling 3 (SOCS-3), obesity (ob) and insulin-like growth factor I (IGF-I), which are involved in adipose metabolism or muscle development, were analyzed. Total RNA was extracted from dorsal subcutaneous adipose tissue and longissimus of 8-month-old Bamei and Largewhite pigs. Semiquantitative reverse transcription-polymerase chain reaction was used to determine the expression levels of the SOCS-3 and ob genes in adipose tissue, and SOCS-3 and IGF-I genes in muscle tissue. The results showed that in adipose tissue the expression level of SOCS-3 was significantly higher in Bamei (obese) pigs than that in Largewhite (lean) pigs (P<0.01). However, in muscle tissue it was significantly lower in Bamei than that in Largewhite pigs (P<0.01). Furthermore, the expression of SOCS-3 was positively correlated to that of ob in adipose tissue and that of IGF-I in muscle tissue. These findings suggest that the difference in SOCS-3 gene expression levels in adipose and muscle tissues, the relationship between SOCS-3 and ob in adipose tissue, and that between SOCS-3 and IGF-I in muscle tissue, might contribute to the different fat deposition and muscle development ability between obese and lean pigs. PMID- 17417688 TI - Central vestibular disorders. AB - Dizziness or vertigo is an erroneous perception of selfmotion or object-motion as well as an unpleasant distortion of static gravitational orientation. It is caused by a mismatch between the vestibular, visual, and somatosensory systems. Thanks to their functional overlap, the three systems are able to compensate, in part, for each other's deficiencies. Thus, vertigo is not a well-defined disease entity, but rather a multisensory syndrome that results when there is a pathological dysfunction of any of the stabilizing sensory systems (e.g., central vestibular disorders, peripheral vestibular diseases with asymmetric input into the vestibular nuclei). This article provides an overview of the most important and frequent forms of central vestibular vertigo syndromes, including basilar/vestibular migraine, which are characterized by ocular motor, postural, and perceptual signs. In a simple clinical classification they can be separated according to the three major planes of action of the vestibulo-ocular reflex: yaw, roll, and pitch. A tonic imbalance in yaw is characterized by horizontal nystagmus, lateropulsion of the eyes, past-pointing, rotational and lateral body falls, and lateral deviation of the perceived straight-ahead. A tonic imbalance in roll is defined by torsional nystagmus, skew deviation, ocular torsion, tilts of head, body, and the perceived vertical. Finally, a tonic imbalance in pitch can be characterized by some forms of upbeat or downbeat nystagmus, fore-aft tilts and falls, and vertical deviation of the perceived straight ahead. The thus defined syndromes allow for a precise topographic diagnosis as regards their level and side. PMID- 17417689 TI - The peptidoglycan recognition proteins LCa and LCx. AB - Infection of bacteria triggers innate immune defense reactions in Drosophila. So far, the only bacterial component known to be recognized by the insect innate immune system is peptidoglycan, one of the most abundant constituents of the bacterial cell wall. Insects use peptidoglycan recognition proteins to detect peptidoglycan and to activate innate immune responses. Such specialized peptidoglycan receptors appear to have evolved from phage enzymes that hydrolyze bacterial cell walls. They are able to bind specific peptidoglycan molecules with distinct chemical moieties and activate innate immune pathways by interacting with other signaling proteins. Recent X-ray crystallographic studies of the peptidoglycan recognition proteins LCa, and LCx bound to peptidoglycan have provided structural insights into recognition of peptidoglycan and activation of innate immunity in insects. PMID- 17417690 TI - Regulation of the NADPH oxidase activity and anti-microbial function of neutrophils by arachidonic acid. AB - Arachidonic acid (AA), a second-messenger molecule released from membrane phospholipids by phospholipase A(2) in activated cells, is a stimulator of neutrophil responses, including the oxygen-dependent respiratory burst. The polyunsaturated fatty acid is also the precursor of biologically active eicosanoids. There are several mechanisms by which AA stimulates the respiratory burst. These include the direct binding of AA to S100 proteins which regulate the assembly of the NADPH oxidase as well as the activation of key signaling molecules which control the respiratory burst. Arachidonic acid also stimulates it own release from membrane phospholipids and this contributes to optimal respiratory burst activity. Thus, increased levels of AA at sites of inflammation will influence the magnitude and course of the inflammatory response, not only by directly affecting the function of infiltrating neutrophils and other leukocytes, but also through its metabolites generated by lipoxygenases and cyclooxygenases. PMID- 17417691 TI - Cytokine production by human leukocytes with different expressions of natural antiviral immunity and the effect of antibodies against interferons and TNF alpha. AB - INTRODUCTION: Two activities of innate antiviral immunity were studied: the resistance of human peripheral blood mononuclear cells (PMBCs) ex vivo to viral infection and the production of cytokines. MATERIALS AND METHODS: Samples of blood were taken from healthy blood donors and from persons with frequent infections of the upper respiratory system. PMBCs were isolated by gradient centrifugation. Vesicular stomatitis virus (VSV) was used as the indicatory virus to infect PMBCs. The cytokines: IFN, TNF, and IL-6 were titrated by biological methods and IL-10 by ELISA. RESULTS: Blood donors were divided for two groups: those with VSV-resistant and those with VSV-sensitive PMBCs and secretion of cytokines by them was compared. The resistant PMBCs produced more cytokines than the sensitive ones. A statistically significant difference, was found only in the case of the IFNs. To examine the contribution of IFNs and TNF in maintaining resistance, leukocytes from both groups were treated with specific anti-cytokine antibodies. The authors' previous study showed that the elimination of spontaneous IFN-alpha, IFN-beta, IFN-gamma, and TNF-alpha from resistant leukocytes resulted in increased VSV replication This indicates the important role of cytokines. In VSV-sensitive PMBCs, anti-IFN-alpha showed the opposite effect (decreased virus replication). In the absence of spontaneous IFN-alpha, disturbances in cytokine production were observed. CONCLUSIONS: Complete resistance of PMBC to VSV infection is accompanied by higher cytokine release, The paradoxical effect of anti-IFN-alpha on virus replication in leukocytes sensitive to viral infection may be attributed to changes in the cytokine profile balance, i.e. high TNF production by VSV-infected leukocytes and a complete reduction of IL-6 production. PMID- 17417693 TI - Do Langerhans cells play a role in vulvar epithelium resistance to squamous cell carcinoma? AB - INTRODUCTION: Langerhans cells (LCs) are a very important part of the skin immune system. MATERIALS AND METHODS: Skin biopsies taken from 13 women after the removal of vulvar squamous cell carcinoma (SCC) who had not been treated earlier for any vulvar diseases were investigated. The control group consisted of 12 women who underwent a plastic surgical operation of the vulva region. Immunohistochemical staining was performed on formalin-fixed, paraffin-embedded tissues samples using antihuman CD1a antibody (NCL-CD1a-235, Novocastra). RESULTS: This study showed a large decrease in LCs in vulvar SCC. CONCLUSIONS: It is postulated that the reduction in the number of LCs may be one of the reasons for a higher tendency of carcinogenesis in the vulvar region. Their role as a main element of the skin immune system in the initiation of this process needs further investigation. It is possible that research on LCs in the skin will cast a new light on their role and even contribute to the prophylaxis and treatment of skin and mucosa carcinomas. PMID- 17417692 TI - Glucose transport in human peripheral blood lymphocytes influenced by type 2 diabetes mellitus. AB - INTRODUCTION: The objective of this study was to evaluate glucose transport into lymphocytes in healthy subjects and patients with type 2 diabetes mellitus (DM) treated either with diet only or with insulin and to propose peripheral blood lymphocytes as a convenient model for cellular glucose transport studies. MATERIALS AND METHODS: Sixty subjects with type 2 DM, 30 treated with diet only and 30 with insulin, were investigated. Thirty healthy subjects matched for age, weight, and sex served as a control group. Deoxy-D-glucose, 2-[(3)H(G)] transport was studied in isolated peripheral blood lymphocytes. Expression of glucose transporters was ascertained by immunocytochemical identification and by Western blotting. RESULTS: In lymphocytes from the control group, deoxy-D-glucose uptake increased gradually with the duration of the experiment. In diabetics treated with insulin, the maximal increase in deoxy-D-glucose uptake was observed after 30 min of the investigation, followed by a plateau phase. In diabetics treated with diet, deoxy-D-glucose uptake increased slowly during the first 30 min. The presence of GLUT1 and GLUT3 in lymphocytes was confirmed in this study. CONCLUSIONS: Glucose transport into lymphocytes is altered in type 2 DM. In lymphocytes from diabetics, the dynamics of deoxy-D-glucose uptake significantly differed from that in healthy subjects. There was also a significant difference between the diabetic groups, representing different modes of therapy and stages of the disease. Glucose transport into lymphocytes is apparently influenced by DM as well as by the mode of therapy. We suggest that peripheral blood lymphocytes may become a promising model for studies on glucose transport in diabetes. PMID- 17417694 TI - Characterization of new glycolipid biosurfactants, tri-acylated mannosylerythritol lipids, produced by Pseudozyma yeasts. AB - Mannosylerythritol lipids (MELs) are glycolipid biosurfactants produced by Pseudozyma yeasts. They show not only the excellent interfacial properties but also versatile biochemical actions. In the course of MEL production from soybean oil by P. antarctica and P. rugulosa, some new extracellular glycolipids (more hydrophobic than the previously reported di-acylated MELs) were found in the culture medium. The most hydrophobic one was identified as 1-O-alka(e)noyl-4-O [(4',6'-di-O-acetyl-2',3'-di-O-alka(e)noyl)-beta-D-mannopyranosyl]-D-erythritol, namely tri-acylated MEL. Others were tri-acylated MELs bearing only one acetyl group. The tri-acylated MEL could be prepared by the lipase-catalyzed esterification of a di-acylated MEL with oleic acid implying that the new glycolipids are synthesized from di-acylated MELs in the culture medium containing the residual fatty acids. PMID- 17417695 TI - Immobilization of polyphenol oxidase on chitosan-SiO2 gel for removal of aqueous phenol. AB - A partially purified potato polyphenol oxidase (PPO) was immobilized in a cross linked chitosan-SiO2 gel and used to treat phenol solutions. Under optimized conditions (formaldehyde 20 mg/ml, PPO 4 mg/ml and pH 7.0), the activity of immobilized PPO was 1370 U/g and its Km value for catechol was 12 mM at 25 degrees C. The highest activity of immobilized enzyme was at pH 7.4. Immobilization stabilized the enzyme with 73 and 58% retention of activity after 10 and 20 days, respectively, at 30 degrees C whereas most of the free enzyme was inactive after 7 days. The efficiency of removing phenol (10 mg phenol/l) by the immobilized PPO was 86%, and about 60% removal efficiency was retained after five recycles. The immobilized PPO may thus be a useful for removing phenolic compounds from industrial waste-waters. PMID- 17417696 TI - Unique effect of nebivolol on coronary hemodynamics: it's all a matter of flow. PMID- 17417697 TI - Statin pleiotropy: a popular concept but clinical significance unclear. PMID- 17417698 TI - Molecular evidence and sequence analysis of a natural reassortant between cucumber mosaic virus subgroup IA and II strains. AB - Cucumber mosaic virus (CMV) is a tripartite RNA virus and has been divided into three subgroups, named IA, IB, and II. Some studies have found a few natural reassortants between CMV subgroups, although reassortment between CMV subgroups is infrequent. In our present work, a CMV reassortant, named CMV-Tsh, was obtained from a tomato plant. The complete sequence of CMV-Tsh genomic RNAs has been determined and analyzed. The results of sequence comparisons and phylogenetic analyses revealed that CMV-Tsh RNAs 1 and 3 are derived from one or two CMV subgroup II strain(s), while RNA2 is derived from a CMV subgroup IA strain. A PCR and restriction enzyme analysis-based method was developed to analyze the possibility of mixed infection by CMV strains of different subgroup in the CMV-Tsh-infected tomato plant. The results of the restriction enzyme analysis proved that CMV-Tsh is the unique strain in the tomato plant. Taken together, CMV-Tsh is a natural reassortant having CMV subgroup IA RNA2 and subgroup II RNAs 1 and 3. PMID- 17417699 TI - Pentachlorophenol residues in suspended particulate matter and sediments from the Yangtze River catchment of Wuhan, China. PMID- 17417700 TI - Treatment with zoledronic acid ameliorates negative geometric changes in the proximal femur following acute spinal cord injury. AB - Acute spinal cord injury is associated with rapid bone loss and an increased risk of fracture. In this double-blind, randomized, placebo-controlled trial, 17 patients were followed for 1 year after administration of either 4 or 5 mg of zoledronic acid or placebo. Bone mineral density (BMD) and structural analyses of the proximal femur were performed using the hip structural analysis program at entry, 6 months, and 12 months. The 17 subjects completed 12 months of observation, nine receiving placebo and eight zoledronic acid. The placebo group showed a decrease in BMD, cross-sectional area, and section modulus and an increase in buckling ratio at each proximal femur site at 6 and 12 months. Six months after zoledronic acid, BMD, cross-sectional area, and section modulus increased at the femoral neck and intertrochanteric regions and buckling ratio decreased consistent with improved bone stability. However, at 12 months, the femoral narrow-neck values declined to baseline. In contrast to placebo, the intertrochanteric region and femur shaft were maintained at or near baseline through 12 months in the zoledronic acid-treated group. Urine N-telopeptide excretion was increased at baseline and declined in both the placebo and treatment groups during the 12 months of observation. We conclude that a single administration of zoledronic acid will ameliorate bone loss and maintain parameters of bone strength at the three proximal femur sites for 6 months and at the femur intertrochanteric and shaft sites for 12 months. PMID- 17417701 TI - Secondary transport of amino acids in prokaryotes. AB - Amino acid transport is a ubiquitous phenomenon and serves a variety of functions in prokaryotes, including supply of carbon and nitrogen for catabolic and anabolic processes, pH homeostasis, osmoprotection, virulence, detoxification, signal transduction and generation of electrochemical ion gradients. Many of the participating proteins have eukaryotic relatives and are successfully used as model systems for exploration of transporter structure and function. Distribution, physiological roles, functional properties, and structure-function relationships of prokaryotic alpha-amino acid transporters are discussed. PMID- 17417702 TI - Mechanism and putative structure of B(0)-like neutral amino acid transporters. AB - The Na(+)-dependent transport of neutral amino acids in epithelial cells and neurons is mediated by B(0)-type neutral amino acid transporters. Two B(0)-type amino acid transporters have been identified in the neurotransmitter transporter family SLC6, namely B(0)AT1 (SLC6A19) and B(0)AT2 (SLC6A15). In contrast to other members of this family, B(0)-like transporters are chloride-independent. B(0)AT1 and B(0)AT2 preferentially bind the substrate prior to the Na(+)-ion. The Na(+) concentration affects the K ( m ) of the substrate and vice versa. A kinetic scheme is proposed that is consistent with the experimental data. An overlapping binding site of substrate and cosubstrate has been demonstrated in the bacterial orthologue LeuT( Aa ) from Aquifex aeolicus, which elegantly explains the mutual effect of substrate and cosubstrate on each other's K ( m )-value. LeuT( Aa ) is sequence-related to transporters of the SLC6 family, allowing homology modeling of B(0)-like transporters along its structure. PMID- 17417703 TI - Serotonin transporters--structure and function. PMID- 17417704 TI - Structure and function of sodium-coupled GABA and glutamate transporters. AB - Neurotransmitter transporters are key elements in the termination of the synaptic actions of the neurotransmitters. They use the energy stored in the electrochemical ion gradients across the plasma membrane of neurons and glial cells for uphill transport of the transmitters into the cells surrounding the synapse. Therefore specific transporter inhibitors can potentially be used as novel drugs for neurological disease. Sodium-coupled neurotransmitter transporters belong to either of two distinct families. The glutamate transporters belong to the SLC1 family, whereas the transporters of the other neurotransmitters belong to the SLC6 family. An exciting and recent development is the emergence of the first high-resolution structures of archeal and bacterial members belonging to these two families. In this review the functional results on prototypes of the two families, the GABA transporter GAT-1 and the glutamate transporters GLT-1 and EAAC1, are described and discussed within the perspective provided by the novel structures. PMID- 17417705 TI - Molecular modeling of PepT1--towards a structure. AB - The proton-coupled uptake of di- and tri-peptides is the major route of dietary nitrogen absorption in the intestine and of reabsorption of filtered protein in the kidney. In addition, the transporters involved, PepT1 (SLC15a1) and PepT2 (SLC15a2), are responsible for the uptake and tissue distribution of a wide range of pharmaceutically important compounds, including beta-lactam antibiotics, angiotensin-converting enzyme inhibitors, anti-cancer and anti-viral drugs. PepT1 and PepT2 are large proteins, with over 700 amino acids, and to date there are no reports of their crystal structures, nor of those of related proteins from lower organisms. Therefore there is virtually no information about the protein 3-D structure, although computer-based approaches have been used to both model the transmembrane domain (TM) layout and to produce a substrate binding template. These models will be discussed, and a new one proposed from homology modeling rabbit PepT1 to the recently crystallized bacterial transporters LacY and GlpT. Understanding the mechanism by which PepT1 and PepT2 bind and transport their substrates is of great interest to researchers, both in academia and in the pharmaceutical industries. PMID- 17417706 TI - Structure and function of cationic amino acid transporters (CATs). AB - The CAT proteins (CAT for cationic amino acid transporter) are amongst the first mammalian amino acid transporters identified on the molecular level and seem to be the major entry path for cationic amino acids in most cells. However, CAT proteins mediate also efflux of their substrates and thus may also deplete cells from cationic amino acids under certain circumstances. The CAT proteins form a subfamily of the solute carrier family 7 (SLC7) that consists of four confirmed transport proteins for cationic amino acids: CAT-1 (SLC7A1), CAT-2A (SLC7A2A), CAT-2B (SLC7A2B), and CAT-3 (SLC7A3). SLC7A4 and SLC7A14 are two related proteins with yet unknown function. One focus of this review lies on structural and functional differences between the different CAT isoforms. The expression of the CAT proteins is highly regulated on the level of transcription, mRNA stability, translation and subcellular localization. Recent advances toward a better understanding of these mechanisms provide a second focus of this review. PMID- 17417708 TI - Toward ribosomal RNA catalytic activity in the absence of protein. AB - The ribosome is the ribonucleoprotein particle responsible for translation of genetic information into proteins. The RNA component of the ribosome has been implicated as the catalytic entity for peptide bond formation based on protease resistance and structural data indicating an all-RNA active site. Nevertheless, peptidyl transfer by ribosomal RNA (rRNA) alone has not been demonstrated. In an attempt to show such activity we generated a minimal construct that comprises much of the 23S rRNA peptidyl transferase center, including the central loop and the A- and P-loops. This minimal rRNA domain was inactive in peptide bond formation under all conditions tested. The RNA was subsequently subjected to six rounds of in vitro selection designed to enrich for this activity. The result was a mutated rRNA sequence that could catalyze the covalent linkage of an A-site and P-site substrate; however, the product did not contain a peptide bond. The current study is an example of an in vitro derived alternate function of rRNA mutants and illustrates the evolutionary possibility that the protoribosome may have used amino acids as substrates before it gained the ability to join them into peptides. Though peptidyl transferase activity in the absence of protein remains elusive, the ease with which alternate catalytic activity was selected from rRNA with a small number of mutations suggests that rRNA may have inherent activity. This study represents a step on the path toward isolating that native activity. PMID- 17417709 TI - Randomized clinical trial comparing two mastectomy techniques. AB - BACKGROUND: The unit was considering the routine use of diathermy scissors for standard mastectomy surgery. We therefore aimed to assess scientifically the outcome of patients following the use of this instrument in their operation before accepting it as a routine procedure. METHODS: A single blind randomized control trial compared the outcome of patients undergoing simple mastectomy using either the standard scalpel blade technique or the bipolar cutting scissors technique. Each arm of the trial contained 30 patients. RESULTS: The two primary outcome measures were blood loss intraoperatively and the operating time. There was a significant difference between the two groups, with a statistically significant benefit in the scissors group in terms of the secondary outcome measures of chest wall clearance and skin flap development as assessments of surgical completeness of mastectomy. There is no evidence of any other secondary outcome measures differing between the treatment groups. CONCLUSION: There is strong evidence that using electric scissors reduces intraoperative blood loss and operating time. There is some evidence that the scissors may provide better surgical completeness of mastectomy. PMID- 17417710 TI - Co-morbidity is a strong predictor of early death and multi-organ system failure among patients with acute pancreatitis. AB - A small but significant percentage of patients with acute pancreatitis die within 2 weeks of hospitalization, usually with multiorgan system failure. To determine the effect of chronic medical comorbidities on early death, we conducted a retrospective analysis of all patients who were hospitalized in California with first-time pancreatitis between 1992 and 2002. Among 84,713 patients, 1514 (1.8%) died within 2 weeks. In a risk-adjusted multivariate model, the strongest predictors of early death were age 65 to 75 years (OR = 2.6, 95% CI: 2.2-3.1 versus <55 years), age over 75 years (OR = 5.2, 95% CI: 4.4-6.1), and the presence of either two chronic comorbid conditions (OR = 3.5, CI: 2.7-4.6) or three or more comorbidities (OR = 7.4, 95% CI: 5.7-9.5). Among the 14,280 patients younger than 55 years who had no chronic comorbid conditions, only 14 (0.1%) died in the first 14 days compared to 701 (5.9%) of 24,852 patients 64 years or older who had three or more comorbidities (RR = 29, 95% CI: 17-50). Comorbid conditions associated with early death included recent cancer, heart failure, renal disease, and liver disease. We conclude that advancing age and the number of chronic comorbid conditions are very strong predictors of early death among patients with acute pancreatitis. PMID- 17417711 TI - A review of resistance patterns and phenotypic changes in gastrointestinal stromal tumors following imatinib mesylate therapy. AB - Gastrointestinal stromal tumors are neoplastic lesions that arise from the interstitial cells of Cajal and are associated with somatic mutations in the tyrosine kinase receptor, KIT. The only known curative therapy is complete surgical resection. Unfortunately, postsurgical recurrence rates exceed 50% and most tumors are resistant to standard chemotherapy and radiation. Imatinib mesylate, a novel tyrosine kinase inhibitor, holds promise as a potential adjuvant therapy to prevent recurrence and improve long-term survival. However, as resistance data emerge, it appears that a potential "escape pathway" may originate from secondary mutations in the KIT receptor. This paper reviews the historical clinical experience with imatinib mesylate and discusses resistance patterns following targeted therapy. We highlight this review with an interesting case report that illustrates unique phenotypic tumoral changes associated with imatinib mesylate resistance. PMID- 17417712 TI - Stomach-preserving distal pancreatectomy with combined resection of the celiac artery: radical procedure for locally advanced cancer of the pancreatic body. AB - To enhance the resectability of cancer of the pancreatic body, a new surgical technique should be developed. Of 25 patients with cancer of the pancreatic body who underwent distal pancreatectomy with curative intent, seven with cancer invasion around the celiac artery underwent stomach-preserving distal pancreatectomy with combined resection of the celiac artery. This procedure secured arterial blood supply to the whole stomach and liver via the inferior pancreaticoduodenal artery without arterial reconstruction. There was no postoperative mortality. One patient developed transient passage disturbance in the duodenum. Another one developed a minor pancreatic fistula. No patients had serious complications related to ischemia of the stomach or liver. The quality of life of the patients after surgery was well maintained, and planned adjuvant therapy was accomplished. Local recurrence was evident in only two patients. The median survival time of patients who underwent distal pancreatectomy with (n = 7) or without (n = 18) resection of the celiac artery was 19 and 25 months, respectively. The overall survival rate was not significantly different between the two groups (P = 0.5300). The present study suggests that this surgical procedure is a rational approach to locally advanced pancreatic body cancer invading around the celiac artery. In view of the feasibility of this procedure, it can also be adopted for less advanced cancer of the pancreatic body to enhance local control and survival. PMID- 17417713 TI - Functional results following elective laparoscopic sigmoidectomy after CT-proven diagnosis of acute diverticulitis evaluation of 43 patients and review of the literature. AB - We performed a prospective study to analyze the functional results following elective laparoscopic sigmoidectomy for computed tomography (CT)-proven diagnosis of acute diverticulitis and review the literature. Forty-three of 45 available patients (96%) who had laparoscopic sigmoidectomy for CT-proven acute diverticulitis answered, after a mean time of 40 months, a questionnaire exploring new abdominal symptoms, bowel function, and the patient's own judgement of the surgical outcome. Surgical technique aimed at removing all the sigmoid by taking down the splenic flexure and do a colorectal anastomosis. Four patients (9%) complained of new abdominal pain. Bowel function was reported as better for 24 patients (56%), unchanged for 16 patients (37%), and worse for 3 (7%). Twenty patients (47%) considered their final result as excellent to good, 17 patients (40%) as satisfying, and 6 patients (13%) as mediocre. Male gender, absence of preoperative history compatible with an irritable bowel syndrome, length of resected sigmoid and residual acute inflammation on histology are statistically predictive of a better postoperative degree of satisfaction. After elective laparoscopic sigmoidectomy for CT-proven diverticulitis, a great majority of patients are very satisfied with their postoperative general comfort. PMID- 17417714 TI - Problem behavior in a community sample of 14- and 19-month-old children: common versus uncommon behaviors, structure, and stability. AB - Few studies have examined the presence, structure, and stability of behavior problems in a community sample of 14- and 19-month-old infants. A questionnaire with items on emotional, attentional, and impulsive behavior and social communication was completed by the parents of 6,491 infants aged 14 months and 1,803 infants aged 19 months. Particularly externalizing behavior problems were reported to present sometimes or often for 50% of more of the sample and could be considered as common. In contrast, social communication problems were reported to be present in less than 10% of the sample. Overall, boys showed more problem behaviors than girls. Principal component analysis at the 14 months data revealed seven factors, which could all be replicated in the 19 months data. Pearson correlations between scores at 14 months and 19 months were highest for oppositional and attention factors (0.68 and 0.63) and lowest for the inhibiton factor (0.38). More than 50% of those scoring in the top 10% for total problem score at 14 months were in the top 10% at 19 months. These results will facilitate the recognition of psychopathology at very early age and the study of its development over time. PMID- 17417715 TI - NMR assignment of the E. coli type 1 pilus protein FimF. PMID- 17417716 TI - Species of Calydiscoides Young, 1969 (Monogenea: Diplectanidae) from lethrinid fishes, with the redescription of all of the type-specimens and the description of C. euzeti n. sp. from Lethrinus rubrioperculatus and L. xanthochilus off New Caledonia. AB - Type-specimens of all known species of Calydiscoides Young, 1969 from lethrinids, namely C. australis Young, 1969 (type-species), C. difficilis (Yamaguti, 1953) Young, 1969, C. duplicostatus (Yamaguti, 1953) Young, 1969, C. rohdei Oliver, 1984 and C. gussevi Oliver, 1984, were re-examined; the male copulatory organs (MCO) of the type-specimens are figured for comparison. Two groups are distinguished within Calydiscoides: the 'difficilis' group, with a massive MCO (C. difficilis and C. rohdei), and the 'australis' group, with a branched MCO (C. australis, C. duplicostatus and C. gussevi). Species of Calydiscoides were found in seven species of Lethrinus (Lethrinidae) off New Caledonia, South Pacific. C. australis and C. gussevi from L. miniatus, their type-host, and C. rohdei from L. atkinsoni, its type-host, are re-described from New Caledonian specimens; all were previously known only from off eastern Australia and are new geographical records for New Caledonia. C. euzeti n. sp. is described from specimens collected from L. rubrioperculatus (type-host) and L. xanthochilus off New Caledonia. Specimens from the two hosts have slightly different measurements of the haptoral hard-parts but a similar morphology of the MCO. The new species belongs to the 'australis' group and is characterised by its MCO, which is 70-83 mum in length, elongate in shape with a characteristic anterior curved whip. PMID- 17417717 TI - Revision of the Grillotia erinaceus (van Beneden, 1858) species complex (Cestoda: Trypanorhyncha), with the description of G. brayi n. sp. AB - Members of the trypanorhynch cestode genus Grillotia Guiart, 1927 belonging to the Grillotia erinaceus (van Beneden, 1858) species complex are redescribed. The type-species of the genus, G. erinaceus, is redescribed from Raja spp. in the eastern and western north Atlantic and the Mediterranean. The redescription establishes the presence of: an hermaphroditic sac; internal and external seminal vesicles (but absence of an accessory seminal vesicle); a uterine pore; and the attachment of the retractor muscle in the mid-region of the tentacular bulb. G. pseuderinaceus Dollfus, 1969 and G. recurvispinis Dollfus, 1969 from Raja spp. in the Mediterranean are considered to be synonyms of G. erinaceus, following Palm (2004). G. dollfusi Carvajal, 1971 from R. chilensis Guichenot off Chile is redescribed from the paratypes and features of the terminal genitalia, consistent with those of G. erinaceus, are described for the first time. G. musculara (Hart, 1936) is redescribed from new material collected from the type-host, R. rhina (Jordan & Gilbert), off Nanaimo on the western coast of Canada. The features of the terminal genitalia of G. musculara are similar to those of the G. erinaceus group. The morphological features of G. borealis Keeney & Campbell, 2001 from Bathyraja spp. in the Bering Sea and the Sea of Okhotsk are summarised and illustrations of this species provided. G. brayi n. sp. is described from Amblyraja radiata from the coasts of Iceland and Britain. The new species differs from other members of the complex in lacking modified hooks 1 and 1' at the base of the tentacle (differentiating it from G. erinaceus), a long pars vaginalis (differentiating it from G. dollfusi) and uncinate hooks in the band on the external surface of the tentacle (differentiating it from G. musculara). Brief descriptions are provided of two apparently new species of Grillotia currently represented in collections by single specimens. PMID- 17417718 TI - Topographical and numerical study of the idiosomal integumentary structures of the larva of four Neotropical species of Amblyomma Koch, 1844 (Acari: Ixodidae). AB - Integumentary structures of the larvae of Amblyomma longirostre (Koch, 1844), A. parvum Aragao, 1908, A. rotundatum Koch, 1844 and from three populations of A. cajennense (Fabricius, 1787) were studied using light microscopy. A new nomenclature for the localisation of the integumentary structures is proposed. Three types of integumentary structures were identified in the larval idiosoma of the four Amblyomma species: lyrifissures, small glands and large wax glands. These structures were observed isolated or associated over the entire idiosoma, except in the scutum, which lacked lyrifisures and large wax glands. Large wax glands were the most stable within and between the tick species, followed by lyrifissures and small glands. Small glands, although relatively stable, showed the highest number of numerical variations within and between the tick species. Even though there were intra-population variations in the topographical and numerical pattern of some integumentary structures of A. cajennense larvae, there was a definitive pattern for most of the specimens, as showed by the similar modal and mean numbers of integumentary structures per tick side. The patterns of lyrifissures, small glands and large wax glands showed little differences when compared between the four Amblyomma species; however, a few differences were well evident. These differences were sufficient to differentiate larvae of the four species. Thus, we expect that the study of integumentary structures on the larvae of other Amblyomma species will be useful in future taxonomic keys for the identification of Amblyomma larvae from the Neotropical region. PMID- 17417719 TI - Functional analysis of Arabidopsis genes involved in mitochondrial iron-sulfur cluster assembly. AB - Machinery for the assembly of the iron-sulfur ([Fe-S]) clusters that function as cofactors in a wide variety of proteins has been identified in microbes, insects, and animals. Homologs of the genes involved in [Fe-S] cluster biogenesis have recently been found in plants, as well, and point to the existence of two distinct systems in these organisms, one located in plastids and one in mitochondria. Here we present the first biochemical confirmation of the activity of two components of the mitochondrial machinery in Arabidopsis, AtNFS1 and AtISU1. Analysis of the expression patterns of the corresponding genes, as well as AtISU2 and AtISU3, and the phenotypes of plants in which these genes are up or down-regulated are consistent with a role for the mitochondrial [Fe-S] assembly system in the maturation of proteins required for normal plant development. PMID- 17417720 TI - Carnitine transporter and holocarboxylase synthetase deficiencies in The Faroe Islands. AB - Carnitine transporter deficiency (CTD) and holocarboxylase synthetase deficiency (HLCSD) are frequent in The Faroe Islands compared to other areas, and treatment is available for both disorders. In order to evaluate the feasibility of neonatal screening in The Faroe Islands we studied detection in the neonatal period by tandem mass spectrometry, carrier frequencies, clinical manifestations, and effect of treatment of CTD and HLCSD. We found 11 patients with CTD from five families and 8 patients with HLCSD from five families. The natural history of both disorders varied extensively among patients, ranging from patients who presumably had died from their disease to asymptomatic individuals. All symptomatic patients responded favourably to supplementation with L: -carnitine (in case of CTD) or biotin (in case of HLCSD), but only if treated early. Estimates of carrier frequency of about 1:20 for both disorders indicate that some enzyme-deficient individuals remain undiagnosed. Prospective and retrospective tandem mass spectrometry (MS/MS) analyses of carnitines from neonatally obtained filter-paper dried blood-spot samples (DBSS) uncovered 8 of 10 individuals with CTD when using both C(0) and C(2) as markers (current algorithm) and 10 of 10 when using only C(0) as marker. MS/MS analysis uncovered 5 of 6 patient with HLCSD. This is the first study to report successful neonatal MS/MS analysis for the diagnosis of HLCSD. We conclude that CTD and HLCSD are relatively frequent in The Faroe Islands and are associated with variable clinical manifestations, and that diagnosis by neonatal screening followed by early therapy will secure a good outcome. PMID- 17417721 TI - Mango profilin: cloning, expression and cross-reactivity with birch pollen profilin Bet v 2. AB - Mango can cause severe anaphylactic reactions. Profilin has been assumed partly responsible for the cross-reactivity between mango fruit and other allergens but has not been finally clarified. In this study, two isoforms of mango fruits profilin were amplified by RT-PCR and 3'RACE from total RNA. Each mango profilin cDNA includes an open reading frame coding for 131 amino acids. The deduced amino acid sequence of the corresponding protein show high identity with other allergenic profilins. Expression of the recombinant mango profilin was carried out in Escherichia coli BL21(DE3) using vector PET28a and the purification of the recombinant protein was performed via affinity chromatography with Ni+ coupled to sepharose. IgE reactivity of recombinant mango profilin was investigated by immunoblot and 8 of 18 mango-allergic patients tested presented specific IgE antibodies to recombinant mango profilin. IgE-inhibition and ELISA inhibition experiments were performed to analyze mango profilin cross-reactivity with profilins from birch pollen and high cross-reactivities have been found. PMID- 17417722 TI - Applying item response theory and computer adaptive testing: the challenges for health outcomes assessment. AB - OBJECTIVES: We review the papers presented at the NCI/DIA conference, to identify areas of controversy and uncertainty, and to highlight those aspects of item response theory (IRT) and computer adaptive testing (CAT) that require theoretical or empirical research in order to justify their application to patient reported outcomes (PROs). BACKGROUND: IRT and CAT offer exciting potential for the development of a new generation of PRO instruments. However, most of the research into these techniques has been in non-healthcare settings, notably in education. Educational tests are very different from PRO instruments, and consequently problematic issues arise when adapting IRT and CAT to healthcare research. RESULTS: Clinical scales differ appreciably from educational tests, and symptoms have characteristics distinctly different from examination questions. This affects the transferring of IRT technology. Particular areas of concern when applying IRT to PROs include inadequate software, difficulties in selecting models and communicating results, insufficient testing of local independence and other assumptions, and a need of guidelines for estimating sample size requirements. Similar concerns apply to differential item functioning (DIF), which is an important application of IRT. Multidimensional IRT is likely to be advantageous only for closely related PRO dimensions. CONCLUSIONS: Although IRT and CAT provide appreciable potential benefits, there is a need for circumspection. Not all PRO scales are necessarily appropriate targets for this methodology. Traditional psychometric methods, and especially qualitative methods, continue to have an important role alongside IRT. Research should be funded to address the specific concerns that have been identified. PMID- 17417723 TI - Effects of expanded cardiac rehabilitation on psychosocial status in coronary artery disease with focus on type D characteristics. AB - Type D personality has been shown to increase the risk for cardiovascular events in patients with coronary artery disease (CAD). We investigated the effects of expanded cardiac rehabilitation on type D score and psychosocial characteristics in 224 CAD patients randomised to either expanded cardiac rehabilitation (stress management, increased physical training, stay at a "Patient Hotel" after discharge and cooking sessions), or routine rehabilitation. Follow-up was 1 year. At baseline patients with a high type D score [patients in the upper quartile of type D score (Q4) i.e., type D patients] had a lower sense of coherence (p < 0.001), a lower quality of life (p < 0.001), more depressive symptoms (p < 0.001) and increased anxiety (p < 0.001) as compared to patients with a low type D score (Q1). During follow-up, type D patients (Q4) randomised to intervention had significant decrements in type D-score (p < 0.01), depression and anxiety (p < 0.05) and an increment in quality of life scores (p < 0.001). Quality of life was also improved in control type D patients (Q4; p < 0.01) but no significant changes were seen in type D score, depression or anxiety. Expanded cardiac rehabilitation reduces type D score, anxiety and depressive symptoms, and improves the quality of life in type D patients. PMID- 17417724 TI - State and trait emotions in delinquent adolescents. AB - OBJECTIVE: To examine the structure of emotions and affective dysregulation in juvenile delinquents. METHOD: Fifty-six juvenile delinquents from a local juvenile hall and 169 subjects from a local high school were recruited for this study. All participants completed psychometric testing for trait emotions followed by measurements of state emotions under two conditions (free association and stress condition). Finally, delinquent participants completed a detailed assessment of past trauma using the Childhood Trauma Interview (CTI). RESULTS: Delinquents exhibit significantly higher levels of negative state and trait emotions when compared to a high school sample. In the delinquent sample chronicity of physical trauma affects the longstanding variable of trait emotionality and severity of trauma, specifically emotional abuse and witnessing violence, shapes negative emotional outcomes in state emotionality. In addition, delinquents appear to experience a wider range of emotions than the comparison sample and were more likely to experience a confluence of state emotions of sadness and anger under stressed conditions. CONCLUSION: Adolescent delinquents appear to have a different experience of negative emotions than comparison adolescents. The experience of emotions appears to differ in state and trait conditions. These emotions may be related to childhood experiences of trauma. PMID- 17417725 TI - Breast-conserving surgery for T3/T4 breast cancer: an analysis of 196 patients. AB - INTRODUCTION: Breast conservation therapy (BCT) increases quality of life and self-esteem of breast cancer patients. In special cancer centers up to 90% of patients are treated with BCT. T3/T4 breast cancer is one of the few contraindications for BCT. However, retrospective data suggest that BCT may be eligible in selected cases of T3/T4 breast cancer. METHOD: We analyzed retrospectively 196 breast cancer patients (operated between 1995 and 2004) suffering from T3/T4 tumors and compared BCT and radiotherapy with mastectomy in these patients in terms of overall survival (OS), local recurrence free-survival (LRFS) and breast cancer-related death (BCRD). RESULT: Demographic data showed no significant differences in prognostic factors between patients treated with mastectomy compared with BCT. Kaplan-Meier curves demonstrated no significant difference for OS, LRFS and BCRD between the two groups. DISCUSSION: Our data strongly suggest that BCT with R0 resection followed by radiotherapy is feasible in patients with T3/T4 breast cancer. Prospective studies have to be performed to further investigate this issue. PMID- 17417726 TI - Astrocytic expressions of phosphorylated Akt, GSK3beta and CREB following an excitotoxic lesion in the mouse hippocampus. AB - Glycogen synthase kinase 3beta (GSK3beta) is believed to play important roles in the regulation of synaptic plasticity, cell survival and circadian rhythms in the mature CNS. However, although several studies have been focused on the GSK3beta, little is known about GSK3beta changes in glial cells under neuropathological conditions. In this study, we evaluated the expressions of molecules associated with the GSK3beta signaling pathway, following the induction of an excitotoxic lesion in mouse brain by kainic acid (KA) injection, which caused pyramidal cell degeneration in the hippocampal CA3 region. In injured hippocampi, Ser47-Akt (protein kinase B, PKB) phosphorylation increased from 4 h until 1 day post injection (PI). Ser9-GSK3beta and Ser133-cAMP responsive element-binding protein (CREB) phosphorylations showed similar spatiotemporal patterns in hippocampi at 1 day until 3 days PI. Double immunohistochemistry also showed that these phosphorylated forms of Akt, GSK3beta and CREB were expressed in astrocytes. For the first time, our data demonstrate the injury-induced astrocytic changes in the levels of phosphorylation of Akt, -GSK3beta and -CREB in vivo, which may reflect mechanisms of glial cells protection or adaptive response to damage. PMID- 17417727 TI - Ischemic colitis associated with segmental arteriovenous malformation mimicking inflammatory bowel disease in a familial adenomatous polyposis patient. PMID- 17417728 TI - Posttraumatic growth, immunity and survival in patients with hepatoma. AB - This study was designed to assess the relationship between posttraumatic growth (PTG), immunity, and survival in patients with biopsy-proven hepatocellular carcinoma (HCC). Forty-one patients with HCC were administered the Posttraumatic Growth Inventory. Total and differential peripheral blood leukocytes (PBL) counts were measured at baseline, 3-, and 6-month follow-up. Survival also was measured from the data of diagnosis to death. Participants who scored above the median for the PTGI total scale score (423 days) survived 186 days longer than participants who scored below the median PTGI total score (237 days). Patients with PTG scores above the median had higher PBL counts. The results of this study suggest that patients with greater PTG scores recover more rapidly from chemotherapy in regards to their white blood cell counts. Further research is warranted regarding the possible immune mediation between PTG and survival. PMID- 17417729 TI - The effect of bariatric surgery on liver histology. PMID- 17417730 TI - Nodular gastritis: an endoscopic indicator of Helicobacter Pylori infection. AB - We prospectively assessed the relationship between nodular gastritis and Helicobacter pylori infection. Of 1409 adults who underwent endoscopy for persistent dyspepsia between June 2004 and August 2005, 41 (2.9%) patients were diagnosed with nodular gastritis (11 [27%] men and 30 [73%] women). The mean age was 45.9 years. A control group of 65 patients without nodular gastritis was also evaluated. The prevalence of H. pylori infection was higher in patients with nodular gastritis than in controls (38/41 [93%] vs. 33/65 [51%]). Of 21 patients treated to eradicate H. pylori, the nodular gastritis pattern resolved or improved in 16 patients on subsequent endoscopy. This study suggests that a nodular pattern of the gastric mucosa on endocscopy is a good indicator for H. pylori infection in adults, with the high positive predictive value of 92.7%. PMID- 17417731 TI - Prescribing patterns and awareness of adverse effects of infliximab: a health survey of gastroenterologists. AB - We sought to determine prescribing patterns and awareness of adverse drug reactions to infliximab among gastroenterologists. A questionnaire was developed and mailed to all gastroenterologists in Maryland and Washington, D.C. Ninety-six of 336 (28.6%) gastroenterologists responded; 86% of respondents use infliximab often or sometimes and 48% infuse infliximab on-site. Only 48% of respondents use immunomodulators prior to infusing infliximab. Thirty-three percent of respondents do not prescribe maintenance infliximab. Respondents reported that infusion reactions occur in 12.9% of infliximab infusions. Most respondents order a purified protein derivative prior to starting infliximab. Respondents underestimated the risk of serious infection, death, demyelinating diseases, and malignancy and overestimated the risk of congestive heart failure. We conclude that a substantial number of gastroenterologists underutilize immunomodulators and fail to prescribe maintenance infliximab. Further, respondents were unaware of the frequency of major adverse events associated with infliximab. Education regarding treatment algorithms in CD and infliximab-related side effects is needed. PMID- 17417732 TI - Heat shock preconditioning induces protein carbonylation and alters antioxidant protection in superficially injured guinea pig gastric mucosa in vitro. AB - According to our previous studies, heat shock preconditioning of gastric mucosa requires modulation of protein synthesis and eicosanoid pathways to induce protection against superficial injury. This may be caused by heat shock-induced oxidative stress. We studied the effect of heat shock preconditioning with normothermic recovery on redox status in superficially injured (1.25 mmol NaCl for 5 min) Ussing chamber perfused guinea pig gastric mucosa allowed to recover for 3 hr after injury. Protein oxidation, lipid peroxidation, level of superoxide dismutase, level of heat shock protein 72 (HSP72), and level of oxygen radical absorbance capacity were measured. Superficial injury increased lipid peroxidation. Heat shock preconditioning decreased oxygen radical absorbance capacity and increased protein carbonyl and HSP72 levels, but inhibited electrophysiologic recovery. Exposure to indomethacin and arachidonic acid (AA) partially abolished this pro-oxidative and inhibitory effect on recovery, but maintained HSP72 levels and decreased protein carbonyls, lipid peroxidation, and oxygen radical absorbance capacity. In conclusion, superficial injury increased lipid peroxidation. Heat shock preconditioning alone induced oxidative stress via indomethacin- and AA-sensitive mechanisms. The development of optimal cytoprotective strategy may therefore require control of oxidative stress and modulation of the eicosanoid pathways. PMID- 17417733 TI - Significance of enhanced expression of nitric oxide syntheses in splenic sinus lining cells in altered portal hemodynamics of idiopathic portal hypertension. AB - Idiopathic portal hypertension (IPH) is characterized by noncirrhotic portal hypertension due mainly to increased intrahepatic, presinusoidal resistance to portal blood flow. Marked splenomegaly is always seen in IPH. To clarify the pathogenetic significance of splenomegaly, immunohistochemical expression of inducible nitric oxide synthese (iNOS), endothelial NOS (eNOS), and endothelin-1 (ET-1) in spleens from patients with IPH was examined. Sinus lining cells of IPH spleens showed diffuse and strong expression of iNOS and eNOS. Sinus lining cells of spleens from patients with liver cirrhosis (LC) also showed positive signals for iNOS and eNOS, but the staining intensity was significantly weak. ET-1 was detectable in only a few mononuclear leukocytes in the red pulp of both IPH and LC spleens. These results suggest that NO liberated in spleen, rather than ET-1, is responsible for the dilatation of splenic sinuses, leading to splenomegaly, and thereby contributes to portal hypertension in IPH. PMID- 17417734 TI - Assessing obesity and other related health problems of mentally ill Hispanic patients in an urban outpatient setting. AB - OBJECTIVE: This paper describes the role of an agency Clinical Director in developing a project to assess and begin to address obesity-related health problems of patients treated in a community-based mental health clinic in New York City. After a five year review of outpatient deaths revealed a high rate of deaths from cardiovascular and diabetes-related issues, the Clinical Director assembled a group of clinicians, researchers, and administrative staff to design a pilot project to assess health and nutrition status of primarily Hispanic day treatment patients with severe and persistent mental illness. METHOD: About 69 of the 105 patients at the clinic were assessed by chart review, interview about nutritional habits and medical care, and somatic measurements for blood pressure, weight, girth, body mass index (BMI), glucose and lipid levels. RESULTS: Patients were predominantly between the ages of 25 and 64 years, 51% were female, and 78% were Hispanic. Around 57% were diagnosed with schizophrenia-spectrum disorders, 86% were receiving antipsychotic medications, and 25% were on two or more antipsychotics. Only 11% of the women and 41% of the men had normal weight. A total of 29% of the women and 18% of the men were overweight (BMI = 25-29.9); and an additional 60% of the women and 41% of the men were obese (BMI > or = 30). Atypical antipsychotic treatment was significantly associated with obesity (BMI > or = 30) (chi sq = 5.5, df = 1, P < 0.025). Using American Heart Association criteria, waist measurements showed significant abdominal obesity among female patients. Blood pressure was elevated in 77% of the patients: 45% were pre hypertensive with BP 120-139/80-89 and 32% were hypertensive with BP > or = 140/90. About 53% had elevated random blood glucoses (>110 mg/dl). On the positive side, patients generally had had recent medical follow-up, and most had adequate cooking facilities. CONCLUSIONS: This project revealed that these predominantly Hispanic, severely mentally ill individuals were at high risk for cardiac illness, highlighting the need for developing culturally-sensitive interventions in urban outpatient psychiatric settings. Findings were disseminated in educational presentations and clinical discussions, and have mobilized an institutional effort to significantly improve medical monitoring for these patients. PMID- 17417736 TI - Neural network analysis of ex-vivo expansion of hematopoietic stem cells. AB - The shortage of hematopoietic stem cells (HSCs) greatly limits their widespread clinical applications. Few studies however, investigated the relationship between the cellular expansion and the influencing factors although wide variety results of the ex-vivo expansion of HSCs existed in literature. Here, a back-propagation (BP) neural network model was employed to evaluate the ex-vivo expansions of nuclear cells (NCs), CD34(+) cells, and colony-forming units (CFU-Cs), where the output was the cellular expansion folds and the inputs include inoculated density, cytokines, resources, serum, stroma, culture time, and bioreactor types. Around 124, 86, and 90 samples were used to train the neural network for the expansion evaluations of NCs, CD34(+ )cells, and CFU-Cs, respectively, while 17, 14, and 10 samples were applied to predict respectively. The results show that for the training of network, the interval accuracy of the expansion folds for the different cells is 85.5, 86.1, and 86.7%, respectively, while the truth-value accuracy is still up to 59.7, 50.0, and 62.2%, respectively within a relative error (RE) of +/-20%. For the prediction of network, the interval accuracy can be up to 82.4, 71.4, and 70%, respectively, while the truth-value accuracy is only 29.4, 14.3, and 50.0%, respectively (RE = +/-20%). Moreover, six verification experiments were carried out based on our interval predicted values and the results proved that the five group predicted conditions lead to the correct expansion of the HSCs with the accuracy more than 80%. Considering the complexity of HSC expansion and complicated wide range of the experimental data, such relatively high interval accuracy for training and prediction as well as verification are satisfied. Therefore this nonlinear modeling makes it possible to describe quantitatively the effects of the culture conditions on the HSC expansion and to predict the optimal culture conditions for higher ex-vivo expansion of HSCs. PMID- 17417737 TI - An in vitro system to evaluate the effects of ischemia on survival of cells used for cell therapy. AB - Maintaining cell viability is a major challenge associated with transplanting cells into ischemic myocardium to restore function. A likely contributor to significant cell death during cardiac cell therapy is hypoxia/anoxia. We developed a system that enabled quantification and association of cell survival with oxygen and nutrient values within in vitro constructs. Myoblasts were suspended in 2% collagen gels in 1 cm diameter x 1 cm deep constructs. At 48 +/- 3 h post-seeding, oxygen levels were measured using microelectrodes and gels were snap-frozen. Bioluminescence metabolite imaging and TUNEL staining were performed on cryosections. Oxygen and glucose consumption and lactate production rates were calculated by fitting data to Fick's second law of diffusion with Michaelis Menten kinetics. Oxygen levels dropped to 0 mmHg and glucose levels dropped from 4.28 to 3.18 mM within the first 2000 mum of construct depth. Cell viability dropped to approximately 40% over that same distance and continued to drop further into the construct. We believe this system provides a reproducible and controllable test bed to compare survival, proliferation, and phenotype of various cell inputs (e.g., myoblasts, mesenchymal stem cells, and cardiac stem cells) and the impact of different treatment regimens on the likelihood of survival of transplanted cells. PMID- 17417738 TI - Prevalence and treatment strategies of dyskinesia in patients with Parkinson's disease. AB - A study into the prevalence and treatment of dyskinesia in Parkinson's disease (PD) patients was performed with 380 PD specialists' completed interviews relating to PD and retrospectively completed 1900 patient record forms for patients with dyskinesia. Physicians reported, that 34% of their PD patients experience dyskinesia, 57% of dyskinetic PD patients were affected by moderately to-completely disabling dyskinesia. Treatment of dyskinesia was looked upon as not satisfactory, fractionating of levodopa dose was used as first choice therapeutic option of dyskinesia. PMID- 17417739 TI - Late-onset frontotemporal dementia associated with a novel PGRN mutation. AB - We describe a new mutation in the PGRN gene (A303AfsX57) associated with late onset frontotemporal dementia and with "cat's eye" shaped intranuclear and cytoplasmatic ubiquitin immunoreactive inclusions in the neuropathological exam. The A303AfsX57 mutation is consistent with a nucleotide deletion in exon 8 (c908delC). This deletion causes a frameshift at codon 303 that introduces a premature termination codon (A303AfsX57). PMID- 17417740 TI - Functional polymorphisms of the 5-HT1A and 5-HT1B receptor are associated with clinical symptoms in migraineurs. AB - Migraine is regarded as a polygenic disease and serotonergic pathways appear to play a major role in its pathogenesis. In the present study, the role of the 5 HT1A and 5-HT1B receptors in migraine was evaluated. The human 5-HT1A receptor gene transcription is modulated by a functional C-1019G promoter polymorphism. The 5-HT1B receptor is the main effector of vasoconstriction in meningeal and cerebral arteries and its functional G861C promoter polymorphism was investigated. We report a positive association of the GG genotype of the 5-HT1A promoter polymorphism with avoidance of physical activity during a migraine attack in comparison to the CC genotype (p = 0.008). Moreover, a positive association of the CC genotype of the G861C polymorphism of the 5-HT1B receptor with the reported intensity of the headache attack on the visual analogue scale was observed (CC 8.3 +/- 1.5 vs. GG 6.9 +/- 1.8; p < 0.05). An association of either polymorphism with migraine with or without aura could not be found. For the first time, our results indicate a role of allelic variation of the 5-HT1A receptor in motion related discomfort in migraineurs and a role of the 5-HT1B receptor polymorphism in headache intensity. PMID- 17417741 TI - Modification of L-DOPA pharmacological activity by MAO inhibitors. AB - Dopamine behaves mainly as a MAO-A substrate in rodent brain, but selective inhibition of MAO-B results in an increased turning activity following L-DOPA administration in hemi-Parkinsonian rodents. Unilateral substantia nigra dopaminergic denervation results in serotonergic hyper-innervation which may increase the contribution of MAO-A in the denervated striatum. Possibly as a result of this, there was no change in striatal MAO-A activity when 95% of dopaminergic innervation was reduced by 6-hydroxydopamine, as assessed by apomorphine-induced turning activity. MAO-B as well as MAO-A may contribute to deamination of dopamine produced from L-DOPA. PMID- 17417742 TI - Testosterone derivatives are neuroprotective agents in experimental diabetic neuropathy. AB - In this study we have assessed the effect of testosterone (T), dihydrotestosterone (DHT) and 5alphaandrostan-3alpha, 17beta-diol (3alpha-diol) therapies on diabetic neuropathy. Diabetes was induced in adult male rats by the injection of streptozotocin and resulted in decreased T and increased 3alpha-diol levels in plasma and in decreased levels of pregnenolone and DHT in the sciatic nerve. Moreover, a reduced expression of the enzyme converting Tinto DHT (i.e., the 5alpha-reductase) also occurs at the level of sciatic nerve, suggesting that the decrease of DHT levels could be due to an impairment of this enzyme. Chronic treatment for 1 month with DHT or 3alpha-diol increased tail nerve conduction velocity and partially counteracted the increase of thermal threshold induced by diabetes. Treatment with DHT increased tibial Na(+),K(+)-ATPase activity and the expression of myelin protein P0 in the sciatic nerve.DHT, 3alpha-diol and T reversed the reduction of intra-epidermal nerve fiber density induced by diabetes. These observations indicate that T metabolites can reverse behavioral, neurophysiological, morphological and biochemical alterations induced by peripheral diabetic neuropathy. PMID- 17417743 TI - Recurrent aseptic meningitis: a new CSF complication of Sjogren's syndrome. PMID- 17417744 TI - Efficacy of anti-coagulant treatment with argatroban on cardioembolic stroke. AB - Argatroban, which is a thrombin inhibitor, has an indication as a treatment in the acute phase on atherosclerotic ischemic stroke in Japan. Howeve, in cardioembolic stroke, argatroban is considered to be contraindicated with the side effect of hemorrhage, though there is no clear clinical evidence to show that argatroban increases hemorrhagic compared with heparin. The efficacy of anticoagulant treatment with argatroban on cardioembolic stroke was evaluated retrospectively in this study. We identified 3,113 patients from the Japan Standard Stroke Registry Study who had had a cardioembolic ischemic stroke. We excluded patients with the anti-platelet treatment or the combination therapy of anticoagulation. Our analyses are therefore based on a cohort of 2,529 patients who were treated either with heparin, and argatroban, or with no anti-coagulation treatment. With multivariable regression, hemorrhagic it was shown that hemorrhage was significantly reduced in heparin and argatroban treatments in the patients with mild severity. There was no significant difference in the recurrence of ischemic stroke between the treatments. Both argatroban and heparin showed dramatic improvement compared with the no treatment standard, but only heparin achieved statistical significance for mortality and change in NIHSS score (admission to discharge) in the moderate stroke subgroup [NIHSS 11-22]. Both heparin and argatroban [more so than heparin alone] have a significantly reduced mortality risk. From the present study, it is suggested that argatroban may be useful on cardioembolic stroke, increasing the improvement of recovery of stroke severity without increasing the risk of hemorrhage. Further prospective studies are awaited for evaluating better the efficacy of argatroban on cardioembolic stroke. PMID- 17417745 TI - Psychogenic tremor and related disorders. AB - Psychogenic tremor is the most common form (55%) of all psychogenic movement disorders. Almost 75% of presenting patients are female. Onset is often abrupt. Preceding events include work related injuries and other accidents. Important clinical characteristic include variability of direction, amplitude and frequency. A positive entrainment test, presence of the co-contraction sign, absence of finger tremor and slowness of voluntary movements are suggestive of psychogenic origin. Co-morbidity with psychiatric disorders is common and includes somatoform disorders (35%), such as pain, diffuse sensory loss, conversion disorders and depression (14%). One fifth (21%) of patients are involved in litigation or compensation issues. Diagnosis is based on history, clinical signs and investigations. A multimodal approach including neuropsychiatric evaluation and psychotherapy should be offered. Prognosis, however, is relatively poor particularly if the condition has persisted for over a year and in the long-term up to 80-90% of patients continue to have abnormal movements. PMID- 17417748 TI - [A word as introduction]. PMID- 17417746 TI - The EU-regulation on medicinal products for paediatric use: impacts on child and adolescent psychiatry and clinical research with minors. AB - BACKGROUND: At present the EU-regulation on medicinal products for paediatric use is in the final legislation phase. The Regulation will bring essential changes to the policy of research with minors, to funding and to regulations of drug development in Europe. METHOD: The article analyses contents of the regulation and possible effects on research with and treatment of mentally ill minors. RESULTS: The regulation seems to be a chance to improve pharmacological treatment for children and bring similar research conditions to Europe as they already exist in the US. Some terms of the regulation must be considered as critical due to vague definitions and ambiguously defined policies in some articles. The designated expert committee will be a powerful institution, but it remains to be seen whether this committee will act in the intended way. It is an existing and real danger that European child and adolescent psychiatry will be neglected by the new regulation, if there is no participation of scientists of this discipline in committees. The regulation makes it necessary for child and adolescent psychiatry to strengthen research in clinical trials and developmental psychopharmacology to get benefits from new legislation and improve health care for mentally ill minors. PMID- 17417749 TI - [Interpretation of serological examinations]. AB - The article summarizes our experience and rules concerning interpretation of serological examination results. It describes in brief the mechanisms of antibody production and the laboratory methods in use. All factors influencing the interpretation of serological examinations are analyzed in detail. Finally, typical situations of application of the serological method in infectious disease diagnostics are mentioned. PMID- 17417750 TI - [Importance of urgent cerebrospinal fluid examination for early diagnosis of central nervous system infections]. AB - To a certain extent, the cerebrospinal fluid (CSF) composition reflects the current status of the central nervous system (CNS). Therefore, studying changes in even the most essential CSF parameters provides enormous scope for obtaining valuable information about processes in the CNS in relation to its disorders. The article aims at presenting our current conception of urgent CSF examination with special emphasis on early diagnosis of central nervous infections. In particular, the focus is on evaluating energy conditions in the CSF compartment and permeability of the blood-brain and blood-CSF barriers, CSF cytology, detecting CNS tissue destruction and bleeding into CNS pathways and monitoring the levels of systemic inflammatory activity. PMID- 17417751 TI - [Value of Western blotting in serodiagnosis of chlamydial infections]. AB - The Western blot (WB) method was verified for serological diagnosis of chlamydial infections. MATERIAL AND METHODS: For testing, sera previously examined by the microimmunofluorescence (MIF) test with either ambiguous results or those suggesting persistent infection were used. RESULTS: Whereas the investigation confirmed adequate sensitivity and specificity of the MIF test for diagnosing Chlamydophila pneumoniae infection, it was less sensitive in case of Chlamydia trachomatis. Long-term persistence of IgA antibodies, detected by the MIF test, was often not confirmed by WB. CONCLUSIONS: The results support the view that detecting antibodies alone, without appropriate clinical symptoms, is not sufficient for antibiotic treatment of any infection. PMID- 17417752 TI - [Syphilis screening and the antiphospholipid syndrome]. AB - Based on literature data and their own experience, the authors recommend in case syphilis screening results are positive for antibodies against a nonspecific antigen and negative for antibodies against a specific antigen to consider this fact a signal of the possible presence of antiphospholipid syndrome. PMID- 17417753 TI - [Guidelines for antiretroviral therapy of human immunodeficiency virus (HIV) infected subjects in the Czech Republic]. AB - HIV infection remains an incurable disease because of the impossibility to eradicate the HIV from the organism. However, the combination antiretroviral therapy (cART) is able to efficiently limit HIV replication and slow down progression of immunodeficiency and thus prolong and improve the quality of HIV+ patients? lives. In HIV(+) pregnant women, the antiretroviral therapy substantially reduces the risk of vertical transmission of the infection. According to present knowledge, the cART is indicated mainly in symptomatic patients with stage B or C diseases and for vertical transmission and postexposure prophylaxis; less clear is the indication of cART for treating acute HIV infection and in asymptomatic patients. Various guidelines for the use of antiretroviral agents issued worldwide, e.g. in the USA, Europe or by the WHO, are not completely identical. The authors present a draft of recommendations for the use of antiretroviral agents in the Czech Republic based on the above mentioned guidelines as well as on their own experience with taking care of HIV/AIDS patients. PMID- 17417754 TI - [Authors' response to M. Giboda's comment on the article Schistosomiasis cases at the Teaching Hospital Kralovske Vinohrady in Prague published in Clinical Microbiology and Infectious Diseases]. PMID- 17417755 TI - Critical care nephrology: can we clone the 'Vicenza Model'? PMID- 17417756 TI - In vitro comparison of two bioartificial liver support systems: MELS CellModule and AMC-BAL. AB - Clinically applied bioartificial liver (BAL) support systems are difficult to compare with regard to overall hepatocyte-specific function and clinical outcome. We compared two clinically applied BAL systems, the Modular Extracorporeal Liver Support (MELS) CellModule and the AMC-bioartificial liver (AMC-BAL) in an in vitro set-up. Both BAL systems were loaded with 10 billion freshly isolated porcine hepatocytes, cultured for 7 days and tested on days 1, 2, 4 and 7. Average decrease in hepatocyte-specific functions over 7 days was 9.7%. Three parameters differed between both bioreactors: lidocaine elimination at days 1 and 2 was significantly higher in the AMCBAL, ammonia elimination showed a significantly higher trend for the AMC-BAL over 7 days and LDH release was significantly lower at day 7 for the MELS CellModule. In conclusion, this first in vitro comparison of two clinically applied BAL systems shows comparable functional capacity over a period of 7 days. PMID- 17417757 TI - The influence of different cultivation conditions on the metabolic functionality of encapsulated primary hepatocytes. AB - The clinical application of bioartificial liver support systems (BALS) is still limited because of technical problems associated with the storage, transport and scale-up of common systems. The encapsulation of primary hepatocytes could solve these problems since the scale-up depends only on the number of the beads and encapsulation leads to protection of the cells during the process of freezing and thawing. Many efforts have been made to find an appropriate material for the encapsulation of primary hepatocytes in terms of mechanical resistance as well as appropriate bio- and hemo-compatibility. This study focuses on the improvement of the metabolic functionality of encapsulated primary hepatocytes. A comparison between two different cultivation models showed that dynamic cultivation conditions lead to a 20.4-fold increase in the albumin production and a 5.21-fold increase in the urea synthesis of encapsulated hepatocytes. Furthermore, the influence of different ratios of the number of the cells to the volume of the media was analyzed. Encapsulated hepatocytes cultured with a high amount of medium were characterized by a significantly higher metabolic activity compared to encapsulated hepatocytes cultured with a low level of medium. Interestingly, the cell concentration per mL alginate has no significant influence on the metabolic activity of encapsulated hepatocytes. In conclusion, different optimization strategies are discussed and, finally, the functionality of encapsulated hepatocytes is compared to the standard model of hepatocyte culture, the collagen sandwich. PMID- 17417758 TI - Correlation between HSP-72 expression and IL-8 secretion in human mesothelial cells. AB - BACKGROUND: Cytotoxicity of peritoneal dialysis fluid (PDF) and peritoneal inflammation are currently regarded as the two major culprits for chronic mesothelial injury and peritoneal membrane failure. In this study, we correlated induction of HSP-72, as a marker of the cellular stress response, to secretion of IL-8, as a marker for pro-inflammatory cytokines, in mesothelial cells upon sublethal PDF exposure. METHODS: Primary omental cell cultures of human mesothelial cells were subjected to sublethal PDF exposure times (CAPD2, Fresenius, Germany). At the end of a 24 hour recovery period, induction of HSP-72 in the cell homogenate and IL-8 secretion in the supernatant was assessed by immunodensitometry and ELISA, respectively. RESULTS: PDF exposure times from 15 min to 60 min resulted in progressively increased HSP-72 expression levels (267 vs 320 vs 419% of controls, p<0.05 vs controls) as well as increased IL-8 secretion (323 vs 528 vs 549% of controls, p<0.05 vs controls) with full cell viability (MTT unchanged to control). HSP-72 expression was statistically significantly correlated with IL-8 secretion. CONCLUSIONS: The significant correlation between HSP-72 expression and IL-8 secretion suggests that the regulation of pro-inflammatory pathways in mesothelial cells exposed to PDF may represent an integral part of their stress response. Future studies to investigate the cellular regulatory mechanism involved are warranted. PMID- 17417759 TI - Nanofibrous poly(epsilon-caprolactone)/poly(vinyl alcohol)/chitosan hybrid scaffolds for bone tissue engineering using mesenchymal stem cells. AB - In the present study, based on a biomimetic approach, novel 3D nanofibrous hybrid scaffolds consisting of poly(epsilon-caprolactone), poly(vinyl alcohol), and chitosan were developed via a multi-jet electrospinning method. The influence of chemical, physical, and structural properties of the scaffolds on the differentiation of mesenchymal stem cells into osteoblasts, and the proliferation of the differentiated cells were investigated. Osteogenically induced cultures revealed that cells were well-attached, penetrated into the construct and were uniformly distributed. The expression of early and late phenotypic markers of osteoblastic differentiation was upregulated in the constructs cultured in osteogenic medium. PMID- 17417760 TI - Interferon therapy for HCV-associated glomerulonephritis: meta-analysis of controlled trials. AB - BACKGROUND: A relationship between hepatitis C virus (HCV) infection and chronic glomerulonephritis (GN) has been asserted on the grounds of epidemiological and experimental data. Although this suggests a role for an antiviral approach to HCV associated GN instead of the more conventional immunosuppressive (or supportive) therapy, the optimal management of HCV related glomerulonephritis remains controversial. OBJECTIVE: To compare antiviral with immunosuppressive therapy for HCV-associated GN. DESIGN: Meta-analysis of controlled clinical trials (CCTs) of the two treatments (antiviral versus immunosuppressive) of HCV-associated GN. METHODS: We used the fixed or random effects model of DerSimonian and Laird, with heterogeneity and sensitivity analyses. The rate of proteinuria and serum creatinine decrease after therapy for HCV-associated GN were regarded as the most reliable outcome end-points. RESULTS: We identified six studies involving 145 unique patients with HCV-associated GN. Pooling of study results demonstrated that proteinuria decreased more commonly after IFN than corticosteroid therapy (OR 1.92 (95% CI, 0.39; 9.57), NS), P-test for heterogeneity, 0.06 (I2=52.9%). In a sensitivity analysis including only CCTs using standard IFN-doses, OR was 3.86 (95% CI, 1.44; 10.33, (P=0.007)), P-test for heterogeneity, 0.18 (I2=35.9%). No improvement of serum creatinine after IFN compared to immunosuppressive therapy was noted (OR, 0.59 (95% CI, 0.21; 1.65), NS), P-test for heterogeneity, 0.76 (I2=0%). Only three CCTs gave information on the rate of proteinuria decrease over follow-up (OR, 5.08 (95% CI, 0.69; 37.31), NS). A few major side effects were noted after IFN administration. CONCLUSIONS: Our meta-analysis indicates that standard IFN-doses were more effective than immunosuppressive therapy in lowering proteinuria of patients with HCV-related glomerulonephritis. However, no significant improvement in serum creatinine was seen by IFN or steroid therapy across the studies. Also, information on proteinuria recurrence after the completion of antiviral therapy was not sufficient. Prospective, randomized trials based on combined antiviral therapy (pegylated IFN plus ribavirin) with adequate dose and follow-up are required to assess the efficacy and safety of antiviral treatment of HCV-associated glomerulonephritis. PMID- 17417761 TI - Continuous renal replacement therapy in neonates and young infants during extracorporeal membrane oxygenation. AB - Extracorporeal membrane oxygenation (ECMO) is a therapy that ensures adequate tissue oxygen delivery in patients suffering cardiac and/or respiratory failure that are unresponsive to conventional therapy. During ECMO, it is common to see a decrease in urine output that may be associated with acute renal failure. In this context, continuous renal replacement therapy (CRRT) should be considered. Our aim is to evaluate a pioneer experience in Latin America, related to the use of CRRT in a group of neonatal-pediatric patients during ECMO. We conducted a retrospective review of patients treated with ECMO at our institution between May 2003 and May 2005. Twelve infants were treated with ECMO, six of them also underwent CRRT. The main reasons for CRRT initiation were fluid overload and progressive azotemia. Observed complications were clots in the filter and excessive ultrafiltration. CRRT was successful in fluid management and solute clearance in all patients. Discharge survival rate was 83%, all of them with normal renal function. Concurrent CRRT with ECMO is technically feasible and efficacious in the management of fluid overload and solute clearance. We report the first experience with these therapies in a Latin American neonatal-pediatric ECMO program associated with the Extracorporeal Life Support Organization. PMID- 17417762 TI - Extracorporeal membrane oxygenation and extracorporeal albumin dialysis in pediatric patients with sepsis and multi-organ dysfunction syndrome. AB - Extracorporeal membrane oxygenation (ECMO) is used in managing patients with potentially reversible cardio-respiratory failure refractory to conventional methods. Multiorgan dysfunction syndrome (MODS), usually due to sepsis, remains the main cause of mortality in such patients. We report a series of six pediatric patients with sepsis-induced MODS where extracorporeal albumin dialysis (EAD) was used while the patients were on ECMO. The age of the patients ranged between 1 month and 17 years. The mean pediatric index of mortality (PIM) score at admission was 67.5%. All these patients further deteriorated with MODS and EAD was used as rescue therapy. At institution of EAD, 4 patients had dysfunction of 4 organs and 2 patients had dysfunction of 5 organs. The number of EAD cycles ranged between 1 and 3. Three out of the 6 patients (50%) survived to discharge from the intensive care unit and two of the six patients (33%) survived to hospital discharge. According to our previous experience and published results, all these patients would have been expected to die. The present results suggest that EAD may prove to have a role in the treatment of pediatric patients with sepsis-induced MODS. Further research is required to identify the group of patients who would benefit most by EAD as well as understand the clearance of inflammatory mediators and other mechanisms involved with the use of EAD. PMID- 17417763 TI - Renal replacement therapy in acute renal failure: which index is best for dialysis dose quantification? AB - BACKGROUND: The "delivered dose" of dialysis may significantly affect the outcome of acute renal failure (ARF) patients requiring dialysis. Our study aimed to elucidate which dose quantification method offers an appropriate parameter to compare different treatments in ARF patients. METHODS: Six sustained low efficiency daily dialysis (SLEDD), and 7 continuous venovenous hemofiltration (CVVH) patients with a prescribed Kt/V of 1.0 were studied during a single treatment. CVVH was studied over the first 24 hours after initiation. SLEDD was performed for 6-12 h. Solute clearance (K) was determined by direct dialysate quantification (DDQ). The single-pool Kt/V (spKt/V), equilibrated Kt/V (eqKt/V), equivalent renal urea clearance (EKRc), and solute removal index (SRI) were calculated. RESULTS: There were no significant differences at enrollment between the SLEDD and the CVVH groups in any patient characteristics except for the serum creatinine levels. The prescribed Kt/V of both groups was similar (SLEDD, 0.9+/ 0.22; CVVH, 1.10+/-0.12, p=NS). The EKRc, which is used to verify kinetic equivalence among patients treated with differing renal replacement therapies (RRT), was higher in CVVH (15.7 in SLEDD; 27.4 in CVVH, p<0.0001), despite the fact that there was no difference between the delivered spKt/V for the SLEDD (1.05+/-0.40) and the CVVH (1.10+/-0.11) groups. The values for SRIurea (0.61 in SLEDD; 1.04 in CVVH, p=0.001), SRIcreatinine (0.55 in SLED; 1.02 in CVVH, p<0.0001), and SRIphosphate (1.81 in SLED; 3.60 in CVVH, p=0.03) were higher in CVVH. The EKRc is calculated assuming a steady state, which is an incorrect assumption in ARF patients with hypercatabolism. CONCLUSION: The SRI calculated using direct dialysate effluent quantification appears to be more reliable as an index of the dialysis dose compared to other methods in ARF patients. However, the use of the dialysate-side SRI is limited by the difficulty of dialysate effluent collection. PMID- 17417764 TI - Citrate anticoagulation during CVVH in high risk bleeding patients. AB - BACKGROUND: Regional citrate anticoagulation (RCA) is an effective form of anticoagulation for continuous renal replacement therapy (CRRT) in patients with contraindications to heparin. Its use has been very limited, possibly because of the need for special infusion solutions and difficult monitoring of the metabolic effects. OBJECTIVE: To investigate the safety and the feasibility of an RCA method for continuous veno-venous hemofiltration (CVVH) using commercially available replacement fluid. METHODS: We evaluated 11 patients at high risk of bleeding, requiring CVVH. RCA was performed using commercially available replacement fluid solutions to maintain adequate acid-base balance. We adjusted the rate of citrate infusion to achieve a post-filter ionized calcium concentration [iCa] <0.4 mmol/L when blood flow was <250 ml/min, or <0.6 mmol/L when blood flow was >250 ml/min. When needed, we infused calcium gluconate to maintain systemic plasma [iCa] within the normal range. RESULTS: Twenty-nine filters ran for a total of 965.5 h. Average filter life was 33.6+/-20.5 h. Asymptomatic hypocalcemia was detected in 6.9% of all samples. No [iCa] values <0.9 mmol/L were observed. Hypercalcemia (1.39+/-0.05 mmol/L) occurred in 2.5% of all samples. We observed hypernatremia (threshold 153 mmol/L) and alkalosis (threshold 7.51) in only 9.3% and 9.4% respectively of all samples, mostly concomitantly. No patient showed any signs of citrate toxicity. CONCLUSIONS: We developed a protocol for RCA during CVVH using commercially available replacement fluid that proved safe, flexible and applicable in an Intensive Care Unit (ICU) setting. PMID- 17417765 TI - Incidence and risk factors of coronary artery disease in patients on chronic hemodialysis. AB - Aim of the study was to estimate the incidence of coronary heart disease (CAD) in patients (pts) with end stage renal disease (ESRD) maintained on chronic hemodialysis (HD) and its association with the presence of predisposing factors. The study included 171 dialysis pts (107 male (M) and 64 female (F)). Mean age of pts was 67+/-13 years, mean time on dialysis 52.7+/-44 months and Body Mass Index (BMI) 25.9+/-3.7 kg/m2. Fifty pts (29.2%) were clinically diagnosed with CAD. The diagnosis was established by coronary angiography in 24 (48%) and in 26 by combined dipyridamole-exercise thallium imaging (52%). Pts' data in association with the development of CAD that were recorded included age, sex, smoking habits, hypertension, obesity, the presence of diabetes mellitus (DM), hyperlipidemia, anemia, low albumin levels, secondary hyperparathyroidism (SHP), the presence of chronic inflammation, as evidenced by the presence of elevated levels of CRP and hyperhomocysteinemia. There was a statistically significant association of increasing age and CAD (p<0.0001). Relative risk (RR) was significantly increased i) in male pts compared to female pts (RR: 8.56, p<0.001), ii) in anemic pts compared to pts with hemoglobin levels< or =11 g/dL (RR: 8.26, p<0.0001), iii) in obese pts compared to pts with BMI < or =30 (RR: 5.09, p<0.005) and iv) in pts with increased levels of homocysteine compared to pts with levels of homocysteine <15 |IM (RR: 4.14, p<0.0001). Using linear regression analysis, CAD was associated with the inadequacy of HD (r = - 0.05, p<0.0001), time on HD (r =0.04, p =0.012) and increasing age (r =0.24, p<0.001). There was no statistically significant association between CAD and the presence of the other traditional risk factors. The incidence of CAD in dialysis pts is significantly increased with age, male sex, obesity, time on dialysis, the presence of anemia, hyperhomocysteinemia and inadequacy of HD. PMID- 17417766 TI - Continuous hemofiltration dose calculation in a newborn patient with congenital heart disease and preoperative renal failure. AB - OBJECTIVE: To report a case of a newborn patient with renal failure due to polycystic kidneys requiring renal replacement therapy, and total anomalous pulmonary venous return requiring major cardiosurgical intervention. SETTING: Pediatric cardiosurgery operatory room and pediatric cardiologic intensive care. PATIENT: A 6-day-old newborn child weighing 3.1 kg. RESULTS: Renal function (creatinine value and urine output) was monitored during the course of the operation and intraoperative renal replacement therapy was not initiated. Serum creatinine concentration decreased from 4.4 to 3 mg/dL at cardiopulmonary bypass (CPB) start and to 1.5 at the end of surgery: the creatinine decrease was provided by the dilutional effect of CPB priming and the infusion of fresh blood from transfusions together with an adequate filtration rate (800 m/L in about 120 minutes). After the operation, extracorporeal membrane oxygenation (ECMO) for ventricular dysfunction and continuous hemofiltration for anuria refractory to medical therapy were prescribed. The hemofiltration machine was set in parallel with the ECMO machine at a blood flow rate of 60 ml/min and a predilution replacement solution infusion of 600 ml/h (4.5 ml/min of creatinine clearance once adjusted on extracorporeal circuits; 3000 mL/m2 hemofiltration): after a single hemofiltration session lasting 96 hours, serum creatinine reached optimal steady state levels around 0.5 mg/dL on postoperative day 2 and 3. CONCLUSION: Administration of intraoperative continuous hemofiltration is not mandatory in the case of a 3-kg newborn patient with established renal failure needing major cardiosurgery: hemodilution secondary to CPB, transfusion of hemoderivates, and optimal UF rate appear to be effective methods for achieving solute removal. If postoperative continuous hemofiltration is started, however, a "dialytic dose" of 4.5 ml/min allows an adequate creatinine clearance, quick achievement of a steady state of serum creatinine concentration and an eventual acceptable rate of inflammatory mediator removal. PMID- 17417768 TI - The evolution of knowledge in medicine (speech given at the University of Kosice- Slovak Republic--on October 6, 2006 on receiving honorary degree in medicine). PMID- 17417770 TI - Abstracts from the American Society of Pediatric Hematology/Oncology (ASPHO) 20th Annual Meeting. May 3-6, 2007. Toronto, Canada. PMID- 17417769 TI - 'Do not dialyze': hits the nail on the head, but ... PMID- 17417771 TI - Histone deacetylase inhibitor, suberoylanilide hydroxamic acid (Vorinostat, SAHA) profoundly inhibits the growth of human pancreatic cancer cells. AB - Tumor suppressor genes are often silenced in human cancer; this can occur by transcriptional repression by deacetylation in the promoter regions, mediated by histone deacetylase (HDAC). HDAC inhibitors can block cancer cell growth by restoring expression of tumor suppressor genes. In this study, we investigated the effects of a HDAC inhibitor, suberoylanilide hydroxamic acid (SAHA) on pancreatic cancer cells. SAHA inhibited the growth of 6 pancreatic cancer cell lines in a dose-dependent manner as measured by MTT and clonogenic assays (ED(50) approximately 10(-6) M) associated with induction of apoptosis, G2 cell cycle arrest and also induced differentiation as indicated by morphology and increased expression of cytokeratin 7. It increased expression of p21(WAF1) (independent of the mutational status of p53), C/EBPalpha, RARalpha and E-cadherin; these genes have been associated with decreased proliferation in other cancers. SAHA decreased cyclin B1 expression; this cyclin normally promotes progression through G2 of the cell cycle. SAHA mediated acetylation of histone H3 globally, as well as, associated with the p21(WAF1) promoter, as measured by chromatin immunoprecipitation. SAHA also decreased levels of c-myc and cyclin D1, independent of an active beta-catenin pathway. In further studies, the combination of SAHA and an inhibitor of DNA methylation, 5-Aza-2'-deoxycytidine, had an enhanced antiproliferative effect on pancreatic cancer cells. In summary, SAHA inhibited the growth of human pancreatic cancer cells by inducing apoptosis, differentiation and cell cycle arrest, as well as increase in the expression of several tumor suppressor genes. SAHA is a novel, promising therapeutic agent for human pancreatic cancers. PMID- 17417772 TI - Resident stromal cell-derived MMP-9 promotes the growth of colorectal metastases in the liver microenvironment. AB - Colorectal cancer, the second leading cause of cancer deaths in the United States, has a high probability of metastasizing to the liver. Matrix metalloproteinases (MMPs) are a family of zinc dependent endopeptidases that are implicated in cancer metastasis and many aspects of tumor progression. Using a splenic injection experimental metastasis model, mice that are genetically deficient in MMP-9 demonstrated a nearly 2-fold decrease in liver weight compared with wild type (WT) mice following injection with MC38 syngeneic colorectal cancer cells. Bioluminesence imaging data demonstrates an early negative effect on tumor growth in MMP-9 null mice when compared with WT controls. Reconstitution of the bone marrow in MMP-9-/- mice with cells competent to produce MMP-9 did not recapitulate the WT phenotype of overwhelming burden of metastatic disease in the liver. In situ hybridization revealed the presence of MMP-9 mRNA in both MC38 tumor cells and in surrounding stromal cells, and immunostaining with anti MMP-9 was consistent with MMP-9 expression by resident liver Kupffer cells. Stromal derived MMP-9 contributes to the establishment and growth of colorectal metastases in the liver. Stromal MMP-9 was derived from resident cells, most likely Kupffer cells, as opposed to infiltrating bone marrow-derived cells and the effect of stromal MMP-9 was independent of expression of MMP-9 by tumor cells. Stromal-derived MMP-9 may represent an important target for selective inhibition in the treatment of metastases. (c) 2007 Wiley-Liss, Inc. PMID- 17417773 TI - hTERT inhibits the Epstein-Barr virus lytic cycle and promotes the proliferation of primary B lymphocytes: implications for EBV-driven lymphomagenesis. AB - Transformation of primary B lymphocytes by Epstein-Barr Virus (EBV) requires the establishment of a latent infection, the expression of several latent viral proteins and a sustained telomerase activity. We investigated the interplay between the activation of human telomerase reverse transcriptase (hTERT), the catalytic rate-limiting component of the telomerase complex, and the expression of latent/lytic EBV genes during the establishment of a stably latent EBV infection of normal B lymphocytes. Cell cultures at early passages after EBV infection greatly differed in their timing of hTERT expression and telomerase activation. Induction of hTERT was dependent on the balance between latent and lytic EBV gene expression, being positively associated with a high ratio of latent/lytic isoforms of latent membrane protein 1, and negatively associated with the expression of BZLF1 gene, the main activator of the viral lytic cycle. In turn, hTERT expression was followed by a decrease in EBV lytic gene expression and virus production. Ectopic expression of hTERT in BZLF1-positive B cell cultures resulted in BZLF1 down-regulation, increased resistance to lytic cycle induction, and enhanced in vitro growth properties, whereas hTERT inhibition by siRNA triggered the activation of the EBV lytic cycle. These findings indicate that hTERT contributes by multiple mechanisms to the EBV-driven transformation of B lymphocytes and suggest that hTERT may constitute a therapeutic target for EBV associated B cell lymphomas. PMID- 17417774 TI - In vitro and in vivo evaluation and a case report of intense nanosecond pulsed electric field as a local therapy for human malignancies. AB - When delivered to cells, very short duration, high electric field pulses (nanoelectropulses) induce primarily intracellular events. We present evidence that this emerging modality may have a role as a local cancer therapy. Five hematologic and 16 solid tumor cell lines were pulsed in vitro. Hematologic cells proved particularly sensitive to nanoelectropulses, with more than a 60% decrease in viable cells measured by MTT assay 96 hr after pulsing in 4 of 5 cell lines. In solid tumor cell lines, 10 out of 16 cell lines had more than a 10% decrease in viable cells. AsPC-1, a pancreatic cancer cell line, demonstrated the greatest in vitro sensitivity among solid tumor cell lines, with a 64% decrease in viable cells. When nanoelectropulse therapy was applied to AsPC-1 tumors in athymic nude mice, responses were seen in 4 of 6 tumors, including clinical complete responses in 3 of 6 animals. A single human subject applied nanoelectropulse therapy to his own basal cell carcinoma and had a complete pathologic response. In summary, we demonstrate that electric pulses 20 ns or less kill a wide variety of human cancer cells in vitro, induce tumor regression in vivo, and show efficacy in a single human patient. Therefore, nanoelectropulse therapy deserves further study as a potentially effective cancer therapy. PMID- 17417775 TI - The screening of the second-site suppressor mutations of the common p53 mutants. AB - Second-site suppressor (SSS) mutations in p53 found by random mutagenesis have shown to restore the inactivated function of some tumor-derived p53. To screen novel SSS mutations against common mutant p53s, intragenic second-site (SS) mutations were introduced into mutant p53 cDNA in a comprehensive manner by using a p53 missense mutation library. The resulting mutant p53s with background and SS mutations were assayed for their ability to restore the p53 transactivation function in both yeast and human cell systems. We identified 12 novel SSS mutations including H178Y against a common mutation G245S. Surprisingly, the G245S phenotype is rescued when coexpressed with p53 bearing the H178Y mutation. This result indicated that there is a possibility that intragenic suppressor mutations might restore the protein function in an intermolecular manner. The intermolecular mechanism may lead to novel strategies for restoring inactivated p53 function and tumor suppression in cancer treatment. PMID- 17417776 TI - Human papillomavirus infection in women in Shenzhen City, People's Republic of China, a population typical of recent Chinese urbanisation. AB - Select cancer registries report that cervical cancer is relatively rare in the People's Republic of China, but may not be representative of the entire country. We carried out a survey of human papillomavirus (HPV) prevalence in 3 samples of women, i.e., general population, factory workers, and tertiary sector workers, in Shenzhen City in 2004. All participants were interviewed and offered gynaecological examination. HPV detection in exfoliated cervical cells was performed using a GP5+/6+ PCR-based assay. Overall HPV prevalence was 18.4% among the general population (n = 534), 11.2% among factory workers (n = 269) and 18.8% among tertiary sector workers (n = 224). Corresponding prevalence for high-risk HPV types was 13.5%, 8.2% and 13.8%, respectively. The most commonly found HPV types were HPV16, 52, 58, 31 and 39. HPV prevalence significantly increased with age in the general population, whereas it was highest below age 25 years in tertiary sector workers. Associations of HPV prevalence with indicators of sexual behaviour were stronger among tertiary sector workers than in the other samples of women. High HPV prevalence in all age groups and the appearance of a 'western type' peak in HPV prevalence among young women employed in the tertiary sector raise important questions concerning the real cervical cancer burden, and its control, in urban China. PMID- 17417777 TI - Epstein-Barr virus B95.8 produced in 293 cells shows marked tropism for differentiated primary epithelial cells and reveals interindividual variation in susceptibility to viral infection. AB - Epstein-Barr virus (EBV), a well-characterised B-lymphotropic agent is aetiologically linked to B cell lymphoproliferations, but the spectrum of diseases the virus causes also includes oral hairy leukoplakia, a benign epithelial lesion, as well as carcinomas of the nasopharynx and of the stomach. However, it is still unclear how EBV accesses and transforms primary epithelial cells. Sixteen samples consisting of primary epithelial cells from the sphenoidal sinus or from tonsils were infected with GFP-tagged recombinant B95.8 EBVs produced in the 293 cell line. The rate of infection was assessed by counting GFP positive cells and cells expressing viral proteins. Primary epithelial cells from all samples were found to be sensitive to EBV infection but there was a marked interindividual variation among the tested samples (2-48% positive cells). This suggests heterogeneity in terms of sensitivity to EBV infection in vivo and therefore possibly to EBV-associated diseases of the epithelium. The virus showed a preferential tropism for differentiated epithelial cells (p63 negative, involucrin positive). In all cases, infected cells expressed EBV lytic proteins but also the LMP1 protein. The viral tropism for differentiated cells and the permissivity of these cells for virus replication reproduced in vitro cardinal features of oral hairy leukoplakia. We have identified a source of EBV that shows unusually strong epitheliotropism for primary epithelial cells that will allow detailed analysis of virus-cell interactions during virus infection, replication and virus-mediated transformation. PMID- 17417778 TI - Genetic investigation of DNA-repair pathway genes PMS2, MLH1, MSH2, MSH6, MUTYH, OGG1 and MTH1 in sporadic colon cancer. AB - Mutations in DNA repair genes have previously been identified as causative factors for hereditary nonpolyposis colon cancer (HNPCC). Recent evidence also supports an association between DNA sequence variation in these genes and sporadic colorectal carcinoma (CRC). Genetic investigation of DNA repair genes PMS2, MLH1, MSH2, MSH6, MUTYH, OGG1 and MTH1, as possible susceptibility factors for sporadic CRC, was done using both a haplotype tagging and a candidate (i.e. coding) single nucleotide polymorphism (SNP) approach. Some 1,068 patients with operated CRC (median age at diagnosis: 59 years) were compared to 738 sex-matched control individuals (median age: 67 years). Haplotype tagging SNPs, previously reported risk variants and all known coding SNPs with a minor allele frequency >0.005 were genotyped in PMS2 (N = 10), MLH1 (N = 11), MSH2 (N = 18), MSH6 (N = 15), MUTYH (N = 7), OGG1 (N = 11) and MTH1 (N = 3). No evidence for an association between CRC and any of the 7 genes was detected, neither with the tagging or coding SNPs nor in a sliding window haplotype analysis (all nominal p values >0.05). The previously reported risk variants D132H in MLH1 and R154H in OGG1 were not even observed in the German population. Genetic CRC risk factors so far identified in DNA repair genes seem to be rare and population-specific. Their association with the disease could not be replicated in German CRC samples. It remains to be elucidated by more systematic, large-scale experiments whether common variants in the same genes, but present across populations, represent risk factors for sporadic CRC. PMID- 17417779 TI - Homeobox gene HOP has a potential tumor suppressive activity in human lung cancer. AB - The homeobox containing gene HOP (Homeodomain Only Protein) was identified in the developing heart and lung where it functions downstream of Nkx2.5 and Nkx2.1 to modulate cardiac and lung gene expression. Previously, we found that HOP was downregulated in lung cancer. In this study, we constructed an expression vector containing the full-length cDNA of HOP and transfected it into a lung cancer cell line H2170. Stable transfection led to an increased expression of HOP confirmed by Northern blot analysis. HOP positive transfectants remarkably reduced the growth rate and the ability of anchorage-independent growth in soft agar, and moreover suppressed the tumor formation in nude mice compared to controls. Transient transfection of Nkx2.1 into H2170 resulted in the overexpression of HOP, and correspondingly, siRNA silencing of Nkx2.1 reduced the expression of HOP in lung cancer cells. Treatment with a differentiation modulating agent 5 bromodeoxyuridine (BrdU) led to restoration of HOP expression in a small cell lung cancer cell line H526. In 29 paired primary lung tumor samples, loss of heterozygosity (LOH) analysis was performed by using the 3 microsatellite markers D4S189, D4S231 and D4S392 around the region of chromosome 4q12 where HOP locates. LOH was only found in 4 out 23 cases (17.4%) indicating that allelic loss is a rare genetic event not responsible for the downregulation of HOP in lung cancer. Taken together, our data suggest that HOP is a potential tumor suppressor possibly involved in lung cancer differentiation, and functions downstream of Nkx2.1. PMID- 17417780 TI - A specific inducible nitric oxide synthase inhibitor, ONO-1714 attenuates inflammation-related large bowel carcinogenesis in male Apc(Min/+) mice. AB - It is generally assumed that inflammation influences carcinogenesis. We previously reported that dextran sodium sulfate (DSS) strongly enhances colon carcinogenesis in the Apc(Min/+) mice and the over-expression of inducible nitric oxide synthase (iNOS) contributes to this enhancement. In the current study, we investigated the effect of a selective iNOS inhibitor, ONO-1714 on colitis related colon carcinogenesis in the Apc(Min/+) mouse treated with DSS. Male C57BL/6J Apc(Min/+) and Apc(+/+) mice were exposed to 1% DSS in their drinking water for 7 days. ONO-1714 was given to the mice at a dose level of 50 or 100 ppm in diet for 5 weeks (during the administration of DSS). The tumor inhibitory effects by ONO-1714 were assessed at week 5 by counting the incidence and multiplicity of colonic neoplasms. Additionally, we assessed serum lipid levels and colonic mRNA expression for cyclooxygenase (COX)-2, iNOS, tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta. Feeding with ONO-1714 significantly inhibited the occurrence of colonic adenocarcinoma in a dose dependent manner in the Apc(Min/+) mice. In addition, the treatment with ONO-1714 significantly lowered the serum triglyceride levels and mRNA expression levels of COX-2, TNFalpha and IL-1beta of colonic mucosa in the DSS-treated Apc(Min/+) mice. Neither ONO-1714 nor DSS affected the colonic pathology in the Apc(+/+) mice. Our findings may suggest that ONO-1714 could therefore serve as an effective agent for suppression of colitis-related colon cancer development in the Apc(Min/+) mice. PMID- 17417781 TI - Combined evaluation of Rad51 and ERCC1 expressions for sensitivity to platinum agents in non-small cell lung cancer. AB - DNA repair enzyme expression in tumor cells possibly affects sensitivity to anti cancer agents. The aim of this study was to determine the relationship between expression status of DNA repair enzymes and chemosensitivity in patients with non small cell lung cancer (NSCLC). NSCLC tissues prepared from the surgical specimens of 41 patients were subjected to immunohistochemical analysis for Rad51 and ERCC1 proteins and to a chemosensitivity test using the MTT assay. The relationships between the expression status of the DNA repair enzymes and ex vivo chemosensitivity to various agents were evaluated. A positive expression for Rad51 and ERCC1 was observed in 17 cases (41%) and 20 cases (49%), respectively. The positivity of Rad51 was closely related to a certain histologic type of squamous cell carcinoma and poor differentiation, and the positivity of ERCC1 tended to be related to squamous cell carcinoma. In chemosensitivity tests, sensitivities to CDDP and CBDCA were significantly lower when both 2 enzymes were positive (p = 0.012 and 0.04 in CDDP, 0.014 and 0.03 in CBDCA). Both Rad51 and ERCC1 expressions showed no significant relationship with sensitivities to paclitaxel, etoposide, vinorelbine, gemcitabine, 5-FU, or irinotecan. In conclusion, combined expression of Rad51 and ERCC1 expression is associated with resistance to platinum agents in the ex vivo study of clinical NSCLC, and evaluation of expression status of both DNA repair enzymes would be a predictor for clinical response to platinum-based chemotherapies. PMID- 17417782 TI - Increased health care utilization among long-term cancer survivors compared to the average Dutch population: a population-based study. AB - In the present study, self-reported health care utilization of cancer survivors is compared with those of an age- and gender-matched normative population and predictors of health care utilization are identified. A population-based, cross sectional survey among 1893 long-term survivors of endometrial and prostate cancer and malignant lymphomas (Hodgkin's and non-Hodgkin's) diagnosed between 1989 and 1998 was conducted using the cancer registry of the Comprehensive Cancer Centre South. Cancer survivors visited their general practitioner somewhat more often compared to the age and gender-matched general Dutch population but this effect was not always statistically significant. In addition, they visited their medical specialist significantly more often. Survivors only sporadically (0-3%) visited or required a dietician, sexologist, oncology nurse, pastor, creative therapy or recovery program. Contact with a psychologist, physiotherapist and other cancer survivors took place somewhat more often. Patients visited a medical specialist less often if they were diagnosed with endometrial cancer (OR = 0.2; 95% CI = 0.1-0.5), if they were diagnosed between 10-15 years ago (OR = 0.6; 95% CI = 0.1-0.5) and if they were not married or divorced (OR = 0.5; 95% CI = 0.3 0.9). Contact with a psychologist was related to having a university or college degree (OR = 3.6; 95% CI = 1.3-9.4). Cancer survivors visited their specialist more often compared to the normative population. Changes in health care, such as less administrative work for the specialist and more efficiency, are probably necessary in order to cope adequately with the increasing demand on the system. PMID- 17417783 TI - Second neoplasms after oesophageal cancer. AB - The authors considered the incidence of second neoplasms among 1,672 oesophageal cancers diagnosed between 1974 and 2004 in the Cancer Registries of the Swiss Cantons of Vaud and Neuchatel, and followed-up to 2004. A total of 141 second neoplasms were observed versus 38.5 expected, corresponding to a standardized incidence ratio (SIR) of 3.7 (95% confidence interval: 3.1-4.3). The SIRs were statistically significant for cancers of the oral cavity and pharynx (57.3), larynx (24.3), lung (6.6) and intestines (2.6). The SIRs were higher in subjects diagnosed below age 50 and in the first year after diagnosis. The SIR of upper digestive and respiratory tract neoplasms was higher for oesophageal cancers diagnosed in the upper (87.5) and middle (68.1), as compared with the lower third (19.4). There was no rise of second oral, pharyngeal and laryngeal cancer with advancing age, and their incidence tended indeed to decline from 100/1,000 at age 40-49 to 25/1,000 at age 70-79. There was no tendency to rise with age in the incidence of first oesophageal cancer in subjects who subsequently developed another upper digestive or respiratory tract neoplasm. The excess risks of upper digestive and respiratory tract neoplasms are attributable to increased diagnosis and registration of second neoplasms following a diagnosis of oesophageal cancer, as well as to heavy tobacco and alcohol consumption in oesophageal cancer cases. The absence of rise in incidence with age is also compatible with the existence of a subset of the population of susceptible individuals. PMID- 17417784 TI - hOGG1 Ser326Cys polymorphism and susceptibility to gallbladder cancer in a Chinese population. AB - The human oxoguanine glycosylase 1(hOGG1) gene encodes a DNA glycosylase that is involved in excision repair of 8-OH-dG (8-hydroxy-2-deoxyguanine) from oxidatively-damaged DNA. To determine whether hOGG1 plays a role in the risk for adenocarcinoma of the gallbladder, we tested the association of this polymorphism with gallbladder cancer in a Chinese population-based, case control study of 204 cases and 209 controls. The subjects were genotyped with a polymerase chain reaction-restriction fragment length polymorphism (PCR-RELP) assay. The association between the genetic polymorphism of this gene and risk of the cancer was examined by using a multivariate analysis. We found that the distribution of hOGG1 Ser326Cys genotypes among controls (Ser/Ser, 37.3%; Ser/Cys, 53.6% and Cys/Cys, 9.1%) was significantly different from that among gallbladder cancer cases (Ser/Ser, 43.1%; Ser/Cys, 36.3% and Cys/Cys, 20.6%). Significantly increased risk for gallbladder cancer was both the hOGG1 326Ser/Cys (Odds ratio [OR] = 1.9, 95% confidence interval (CI) = 1.0-3.7) and hOGG1 326Cys/Cys genotypes (OR = 4.5, 95% CI = 1.1-22.4). We observed no statistically significant association between hOGG1 genotype and gallbladder cancer association in gallstone absence. In contrast, a near-significant increase in risk for gallbladder cancer was observed for gallstone presence with the hOGG1 326Ser/Cys genotype (OR = 2.2, CI = 1.4-3.5) whereas a significant increase in association for gallbladder cancer was observed for gallstone presence with the 326Cys/Cys genotype (OR = 6.1, CI = 2.1-27.2). These data corresponded with the fact that a significant trend towards increased association for gallbladder cancer was observed with potentially higher-risk hOGG1 genotypes in gallstone presence(p < 0.001, chi(2) trend test)but not in gallstone absence(p = 0.89, chi(2) trend test). A significant increase in risk for gallbladder cancer was observed for larger gallstone (those with stone diameters 2 cm or greater) with the hOGG1 326Ser/Cys(OR = 1.9, 95% CI = 1.1-2.9) and hOGG1 326Cys/Cys genotypes(OR = 5.9, 95% CI = 1.6-18.0). These data are consistent with the observation that a significant trend towards increased risk for gallbladder cancer was observed with potentially higher-risk hOGG1 genotypes in gallbladder cancer patients with larger gallstone (p < 0.001, chi(2) trend test). However, we observed no statistically significant association between hOGG1 genotype and gallbladder cancer risk in gallbladder cancer patients with smaller gallstone (those with stone diameters 2 cm smaller) (hOGG1 326Ser/Cys:OR = 2.2, 95% CI = 0.8-4.0; hOGG1 326Cys/Cys:OR = 2.9, 95% CI = 0.6-29.4; p = 0.06, chi(2) tread test). These results suggest that hOGG1 Ser326Cys polymorphism is associated with gallbladder cancer risk. PMID- 17417785 TI - High rate of centrosome aberrations and correlation with proliferative activity in patients with untreated B-cell chronic lymphocytic leukemia. AB - B-cell chronic lymphocytic leukemia (CLL) is characterized by a high rate of clonal genomic alterations and a low proliferative activity with cell cycle arrest in G(0)/G(1) phase. Recently, centrosome aberrations have been described as a possible cause of chromosomal instability and aneuploidy in many human malignancies. To investigate whether centrosome aberrations do occur in CLL and whether they correlate with common prognostic factors and disease activity, we examined peripheral blood mononuclear cells (PBMC) from 70 patients with previously untreated CLL using an antibody to gamma-tubulin. All 70 CLL samples displayed significantly more cells with centrosome aberrations (median: 26.0%, range 11.0-41.5%) as compared to peripheral blood B lymphocytes from 20 age matched, healthy individuals (median: 2.0%, range 0-6%; p < 0.001). The extent of centrosome aberrations correlated with the proliferative activity of the CLL cases as measured by lymphocyte doubling time (p = 0.02) as well as with time to first treatment (p = 0.05). Accordingly, more centrosome aberrations were found in PHA-stimulated T lymphocytes from healthy individuals as well as in B cells from surgically removed tonsil tissue of patients with acute tonsillitis as compared to the peripheral blood B lymphocytes from the control group. In contrast, no correlation was observed between centrosome aberrations and immunoglobulin VH gene mutation status or cytogenetically defined risk groups. These findings suggest that, despite the common observation of most CLL cells remaining in G(0)/G(1) phase, their centrosome replication process is deregulated and correlates to the proliferative activity of CLL cells. PMID- 17417786 TI - Enhanced transgenesis by intracytoplasmic injection of envelope-free lentivirus. AB - We demonstrate enhanced transgenesis in mice by intracytoplasmic injection of envelope-free lentivirus. Envelope-free lentivirus carrying the green fluorescent protein (GFP) gene under the control of the ubiquitin promoter (LVU-GFP) was microinjected into the cytoplasm of mouse zygotes prior to embryo transfer. Ninety-seven percent (31/32) of the adult mice were confirmed transgenic by PCR and Southern blot analysis; all founder mice express GFP when tail snips were examined by fluorescent microscopy prior to genomic DNA extraction. Transgene insertion numbers ranging from 1 to 32 were revealed by Southern blot analysis. Germline transmission was confirmed by the presence of transgene in F1 offspring. As expected, a lower transgenic rate (2.2%; 1/46) resulted when envelope-free LVU GFP was microinjected into the perivitelline space (PVS) because cell recognition followed by membrane fusion between the viral envelope and the target cell is prerequisite for successful infection by envelope viruses. Here we demonstrate the competence of envelope-free lentivirus in establishing stable gene integration by germline transgenesis in mice at high efficiency, by intracytoplasmic viral injection (INVI) of envelope-free lentivirus into mouse zygotes. PMID- 17417787 TI - Drosophila Bcl-2 proteins participate in stress-induced apoptosis, but are not required for normal development. AB - Many developing tissues require programmed cell death (PCD) for proper formation. In mice and C. elegans, developmental PCD is regulated by the Bcl-2 family of proteins. Two bcl-2 genes are encoded in the Drosophila genome (debcl/dBorg1/Drob 1/dBok and buffy/dBorg2) and previous RNAi-based studies suggested a requirement for these in embryonic development. However, we report here that, despite the fact that many tissues in fruit flies are shaped by PCD, deletion of the bcl-2 genes does not perturb normal development. We investigated whether the fly bcl-2 genes regulate non-apoptotic processes that require caspases, but found these to be bcl-2 gene-independent. However, irradiation of the mutants demonstrates that DNA damage-induced apoptosis, mediated by Reaper, is blocked by buffy and that debcl is required to inhibit buffy. Our results demonstrate that developmental PCD regulation in the fly does not rely upon the Bcl-2 proteins, but that they provide an added layer of protection in the apoptotic response to stress. PMID- 17417788 TI - Double-inducible gene activation system for caspase 3 and 9 in epidermis. AB - Expression of genes with tight and precise temporal and spatial control is desired in a wide variety of applications ranging from cultured cells and transgenic animals to gene therapy. While current inducible systems, such as RU486 and chemical inducers of dimerization (CID), have improved earlier inducible models (Gossen et al., 1995, Science. 268:1766-1769; Wang et al., 1994, Proc Natl Acad Sci USA 91:8180-8184), no single system is perfect at present. One potential drawback of these systems is leakage of transgene expression, causing limitations of each system. We have developed an inducible model containing both RU486 and CID systems, which in addition to inducing caspase activation, has potential applicability specifically to other genes encoding proteins that require a dimerization event for activation. This Double-Inducible Gene Activation System generates two barriers for the target gene expression and protein activation thereby minimizing leakage. PMID- 17417789 TI - Precursor natural killer cell leukemia. AB - Natural killer (NK) cell tumors are a rare and heterogeneous group of disorders. Immature NK cell tumors are less common, and are less recognized and defined than mature NK cell tumors. There is insufficient experience of diagnosis and treatment with immature NK cell tumors, especially in pediatric patients. Here we describe a pediatric patient with precursor NK cell leukemia and review the literature of the previously reported cases in children to further help characterize the diagnosis, treatment, and outcome. PMID- 17417790 TI - Successful treatment of refractory immune hemolysis following unrelated cord blood transplant with Campath-1H. AB - Immune-mediated hemolytic anemia is a well-recognized complication of hematopoietic stem cell transplantation. We report on a 6-year-old boy with X linked adrenoleukodystrophy who developed severe delayed alloimmune hemolytic anemia associated with immune-mediated neutropenia and thrombocytopenia following major ABO incompatible unrelated cord blood transplantation. The patient's cytopenias were refractory to treatment with corticosteroids, cyclosporine, intravenous immune globulin, rituximab, and pentostatin. After one course of Campath-1H his hematologic parameters normalized, suggesting that the compound may be an effective therapy for complex immunohematologic disorders complicating hematopoietic stem cell transplantation. The case also emphasizes the importance of T-cells in transplant associated immune cytopenias. PMID- 17417791 TI - False positivity of FDG-PET/CT in a child with Hodgkin disease. AB - Role of Positron Emission Tomography (PET) with F-18-2-fluoro-2-deoxy-D-glucose (FDG) in staging of Hodgkin disease is well established despite several controversies. We report a Stage III Hodgkin lymphoma patient with false positive FDG-PET/CT results. Seven-year-old male with Hodgkin lymphoma was in remission at end of chemotherapy. At third and fourth month of postchemotherapy follow-up, increased Gallium uptake and positive FDG-PET/CT in right lower quadrant of abdomen was observed. Open biopsy revealed lymphoid hyperplasia. He has been followed for 21 months without any evidence of disease. Despite its documented benefit, we believe that results of FDG-PET/CT should be interpreted with great caution in order to avoid unnecessary interventions. PMID- 17417792 TI - Helicobacter pylori associated gastric Burkitt lymphoma. AB - Chronic Helicobacter pylori (H. pylori) infection has been linked to lymphoma of gastric mucosa-associated lymphoid tissue (MALT), a tumor that is typically localized at presentation. Sporadic Burkitt lymphoma (BL) frequently presents as an abdominal mass. However, primary gastric BL is unusual and the role of H. pylori in its pathogenesis is unclear. We describe a 12 year old with localized gastric BL and concurrent H. pylori infection. In this report, we discuss the potential association between H. pylori and gastric BL and also highlight certain similarities between gastric MALT lymphomas and BL arising in the stomach. PMID- 17417793 TI - Identification of a crystal cell-specific enhancer of the black cells prophenoloxidase gene in Drosophila. AB - In Drosophila, Black cells (Bc) encodes a Prophenoloxidase and is expressed late in the maturation of crystal cells, which are blood cells involved in wound healing and immune encapsulation. Enhancer analysis of Bc revealed a 1,025-bp upstream sequence that regulates gene expression in a crystal cell exclusive pattern. Expression of this fragment is altered by mutations in the GATA family serpent (srp) and RUNX family lozenge (lz) genes; Srp and Lz are required for crystal cell specification. Deletional analysis uncovered a 330-bp crystal cell specific sequence, which contains two GATA and three Lz binding sites. Mutational analysis revealed that both GATA sites are necessary, but not sufficient for crystal cell expression. However, one of the Lz sites is essential for crystal cell expression. Thus, Srp and Lz do not just specify the crystal cell lineage, but also regulate the later differentiation of these cells. Additionally, we now have a sensitive tool for marking crystal cells in live animals. PMID- 17417794 TI - X-linked dyskeratosis congenita in Malaysia. PMID- 17417795 TI - Proliferative index and CD15 expression in pediatric classical Hodgkin lymphoma. AB - BACKGROUND: This study was conducted to assess the clinical and prognostic significance of lack of CD15 expression, proliferative index (PI), and expression of tumor suppressor protein p53 in pediatric classical Hodgkin lymphoma (CHL). PROCEDURE: Pre-treatment lymph node (LN) biopsies were studied by immunohistochemistry for immunophenotyping of the lymphoma and with Ki-67 (PI) and p53 antibodies. Expression of CD15 antigen on the Hodgkin and Reed-Sternberg (H-RS) cells, proliferation, and apoptosis parameters were correlated with clinical stage, response to chemotherapy alone, overall (OS) and failure-free survival (FFS). RESULTS: One hundred and twenty-one children with CHL were studied. Expression of Ki-67 and p53 in H-RS cells was seen in 100% and 89.9% of the cases, respectively. Loss of CD15 expression, seen in 12 (9.9%) cases, was significantly associated with p53 negativity and was an independent prognostic factor for poor OS and poor FFS. PI CO pi* back-donation in this series. PMID- 17417826 TI - Front waves in the NO + NH3 reaction on Pt{100}. AB - In the present work, we spatially extended a brand new kinetic mechanism of the NO + NH3 reaction on Pt{100} to simulate the experimentally observed spatiotemporal traveling waves. The kinetic mechanism developed by Irurzun, Mola, and Imbihl (IMI model) improves the former model developed by Lombardo, Fink, and Imbihl (LFI model) by replacing several elementary steps to take into account experimental evidence published since the LFI model appeared. The IMI model achieves a better agreement with the experimentally observed dependence of the oscillation period on temperature. In the present work, the IMI model is extended by considering Fickean diffusion and coupling via the gas phase. Traveling waves propagating across the surface are obtained at realistic values of temperature and partial pressure. A transition from amplitude to phase waves is observed, induced either by temperature or by the gas global coupling strength. The traveling waves simulated in the present work are not associated with fixed defects, in agreement with experimental evidence of spiral centers capable of moving on the surface. Also, the IMI model adequately predicts the presence of macroscopic oscillations in the partial pressures of the reactants coexisting with front wave patterns on the surface. PMID- 17417827 TI - An efficient LDA+U based tight binding approach. AB - The functionals usually applied in DFT calculations have deficiencies in describing systems with strongly localized electrons such as transition metals or rare earth (RE) compounds. In this work, we present the self-consistent charge density based functional tight binding (SCC-DFTB) calculation scheme including LDA+U like potentials and apply it for the simulation of RE-doped GaN. DFTB parameters for the simulation of GaN and a selection of rare earth ions, where the f electrons were explicitly included in the valence, have been created. The results of the simulations were tested against experimental data (where present) and against various more sophisticated but computationally more costly DFT calculations. Our approach is found to correctly reproduce the geometry and the energetic of the studied systems. PMID- 17417828 TI - The structure and energetics of 3He and 4He nanodroplets doped with alkaline earth atoms. AB - We present systematic results, based on density functional calculations, for the structure and energetics of 3He and 4He nanodroplets doped with alkaline earth atoms. We predict that alkaline earth atoms from Mg to Ba go to the center of 3He drops, whereas Ca, Sr, and Ba reside in a deep dimple at the surface of 4He drops, and Mg is at their center. For Ca and Sr, the structure of the dimples is shown to be very sensitive to the He-alkaline earth pair potentials used in the calculations. The 5s5p <-- 5s2 transition of strontium atoms attached to helium nanodroplets of either isotope has been probed in absorption experiments. The spectra show that strontium is solvated inside 3He nanodroplets, supporting the calculations. In the light of our findings, we emphasize the relevance of the heavier alkaline earth atoms for analyzing mixed 3He-4He nanodroplets, and in particular, we suggest their use to experimentally probe the 3He-4He interface. PMID- 17417829 TI - Excitation energies and photoabsorption oscillator strengths of the Rydberg series in CF3Cl. A linear response and quantum defect study. AB - Vertical excitation energies of the CF(3)Cl molecule have been obtained from a response function approach with a CC reference function to determine absolute photoabsorption oscillator strengths in the molecular-adapted quantum defect orbital formalism (MQDO). The present work covers more highly excited Rydberg states than have been experimentally reported. Assessing of the reliability of the present calculations is provided through a comparative analysis between the results of the molecule and the Cl atom. This can be used to allow for predictions of the same type of properties in other analogous systems. PMID- 17417830 TI - Hydration of cyclohexylamines: CPCM calculation of hydration Gibbs energy of the conformers. AB - This work presents a theoretical study on the hydration of cyclohexylamine and isomers of cyclohexyldiamine. All possible conformers were fully optimized in solution using the conductor-like polarizable continuum model (CPCM) and density functional theory. Values of the Gibbs energy of solvation, its respective contributions (electrostatic, nonelectrostatic and conformational change), and the relative Gibbs energy of the conformers in aqueous solution and gas phase are reported. From these values and the Boltzmann populations of the conformers in both phases, the weighted mean values of DeltaG(solv) for the compounds are calculated. Three structural features were found to be important for the hydration of these compounds: the distance between the two NH2 groups (proximity disfavors hydration), their position relative to the ring (equatorial is preferred over axial), and the orientation of the nitrogen lone-pairs (gauche is more favorable to hydration than trans). In the particular case of vicinal cyclohexyldiamines, in addition to these two factors, the relative orientation of one group to the other should also be taken into account. PMID- 17417831 TI - A refined pharmacophore identifies potent 4-amino-7-chloroquinoline-based inhibitors of the botulinum neurotoxin serotype A metalloprotease. AB - We previously identified structurally diverse small molecule (non-peptidic) inhibitors (SMNPIs) of the botulinum neurotoxin serotype A (BoNT/A) light chain (LC). Of these, several (including antimalarial drugs) contained a 4-amino-7 chloroquinoline (ACQ) substructure and a separate positive ionizable amine component. The same antimalarials have also been found to interfere with BoNT/A translocation into neurons, via pH elevation of the toxin-mediated endosome. Thus, this structural class of small molecules may serve as dual-function BoNT/A inhibitors. In this study, we used a refined pharmacophore for BoNT/A LC inhibition to identify four new, potent inhibitors of this structural class (IC50's ranged from 3.2 to 17 muM). Molecular docking indicated that the binding modes for the new SMNPIs are consistent with those of other inhibitors that we have identified, further supporting our structure-based pharmacophore. Finally, structural motifs of the new SMNPIs, as well as two structure-based derivatives, were examined for activity, providing valuable information about pharmacophore component contributions to inhibition. PMID- 17417832 TI - Graphical representation and numerical characterization of H5N1 avian flu neuraminidase gene sequence. AB - The high degree of virulence and potential for development of a pandemic strain of the H5N1 avian flu has resulted in wide interest in characterization of the various genes of the H5N1 virus genome. We have considered for our analysis all 173 available complete sequences, as of February 2006, of the neuraminidase gene, which is the target of the most effective treatment regimen comprising the inhibitors oseltamivir and zanamivir. We have used a 2D graphical representation of the neuraminidase RNA sequences of H5N1 strains to identify a few distinct structural motifs. The H5N1 strains were split into two main classes: strains that were benign to human beings in the years up to 1996 and the period 1999-2002 and strains that were highly pathogenic to humans in the periods 1997 and 2003 to present. Comparisons with earlier H1N1 pandemic and epidemic strains have also been made to understand the current status of the gene. Our findings indicate that the base composition and distribution patterns are significantly different in the two periods, and this may be of interest in studying mutational changes in such viral genes. PMID- 17417833 TI - Electrolytes for dye-sensitized solar cells based on interhalogen ionic salts and liquids. AB - In this paper, we report on the preparation of interhalogen ionic liquids of the general formula [K+]XY2-, where K+=1,3-dialkylimidazolium, 1,2,3 trialkylimidazolium, or N-alkylpyridinium; XY2-=IBr2- or I2Br-. These compounds were characterized in solution and the solid state by NMR, IR, Raman, and mass spectroscopy. The crystal structure of the compound [Me2BuIm]IBr2 (7) shows that the IBr2- anion has a linear Br-I-Br structure. Indications of an equilibrium between different forms of XY2- anions in solution are observed. Interhalogen ionic salts and liquids were used as electrolyte components for encapsulated monolithic dye-sensitized solar cells. Overall light-to-electricity conversion efficiencies up to 6.4%, 5.0%, and 2.4% at 1000 W/m2 were achieved by using electrolytes based on interhalogen ionic salts and gamma-butyrolactone, glutaronitrile, or native ionic liquids as solvents, respectively. Moreover, in terms of stability, the cell performance lost 9-14% of the initial performance after 1000 h illumination at 350 W/m2. PMID- 17417834 TI - Defects in divided zinc-copper aluminate spinels: structural features and optical absorption properties. AB - Zn1-xCuxAl2O4 (0 < or = x < 0.30) compounds have been synthesized by polyesterification using metallic salts and annealing at low temperatures as well as by conventional solid state. XRD-powder data refinements (Rietveld method) have demonstrated that both compound series crystallize in the spinel structure (Fd3m) and exhibit similar inversion rates. This low-temperature route lead to metastable phases with crystallite sizes around 40 nm whereas particle sizes are larger than 1 moicrom in the case of solid-state route. This preparative method largely described in the literature allows stabilizing reduced copper states thanks to the presence of reductive organic species, which are decomposed below T = 700 degrees C. The absorption spectra of the x = 0.15 composition exhibit strong differences depending on the synthesis route. These differences can be explained by the occurrence of Cu2+/Cu+ mixed valencies in compounds prepared by the low-temperature route; 33% of monovalent copper has been identified in the x = 0.15 composition prepared by low-temperature process, whereas the solid-state compound contains only divalent copper. Reductive properties of polyesterification reaction implying citric acid and low annealing temperature (T = 700 degrees C) are mainly responsible of the occurrence of the Cu2+/Cu+ mixed valencies. Actually, the annealing under air at T = 1000 degrees C of divided zinc-copper aluminates prepared at low temperatures (T = 700 degrees C) leads to the oxidation reaction Cu+ --> Cu2+ + e- confirmed by the evolution of magnetic measurements, ESR spectra, and optical absorption properties. Defects such as oxygen vacancies in the anionic network leading to reduction in the cations coordination number could also explain the strong evolution of optical absorption spectra especially around lambda = 700 nm where intervalencies transfer (Cu+/Cu2+) as well as intra-atomic d-d transitions (Cu2+ in a 5-fold coordination) can occur. Finally the occurrence of monovalent and divalent copper at the surface of such divided oxides, probably in tetrahedral sites, has been demonstrated by FTIR spectroscopy using the co-adsorption of CO and NO as probe molecules. PMID- 17417835 TI - Oxygenation of simple zinc alkyls: surprising dependence of product distributions on the alkyl substituents and the presence of water. AB - Oxygenation reactions of dialkylzinc solutions have been investigated. Me2Zn reacts with oxygen in the absence of water to give the bis(heterocubane) [(MeZn)6Zn(OMe)8], whereas Et2Zn and iPr2Zn afford the mono(heterocubanes) [(RZn)4(OR)4]. In the presence of small amounts of water (added during or after the oxygenation) the product types are reversed for Me2Zn and Et2Zn giving [(MeZn)4(OMe)4] and [(EtZn)6Zn(OEt)8], while being retained for iPr2Zn (giving [(iPrZn)4(OiPr)4]). Full characterization of all products by NMR spectroscopy, mass spectrometry, and elemental analyses is provided, and crystal structures of [(EtZn)6Zn(OEt)8] and [(iPrZn)4(OiPr)4] are reported. A rationalization of the different reactivities is attempted on the basis of DFT-calculated energies of some key reactants. PMID- 17417836 TI - Comparative study of uranyl(VI) and -(V) carbonato complexes in an aqueous solution. AB - Electrochemical, complexation, and electronic properties of uranyl(VI) and -(V) carbonato complexes in an aqueous Na2CO3 solution have been investigated to define the appropriate conditions for preparing pure uranyl(V) samples and to understand the difference in coordination character between UO22+ and UO2+. Cyclic voltammetry using three different working electrodes of platinum, gold, and glassy carbon has suggested that the electrochemical reaction of uranyl(VI) carbonate species proceeds quasi-reversibly. Electrolysis of UO22+ has been performed in Na2CO3 solutions of more than 0.8 M with a limited pH range of 11.7 < pH < 12.0 using a platinum mesh electrode. It produces a high purity of the uranyl(V) carbonate solution, which has been confirmed to be stable for at least 2 weeks in a sealed glass cuvette. Extended X-ray absorption fine structure (EXAFS) measurements revealed the structural arrangement of uranyl(VI) and -(V) tricarbonato complexes, [UO2(CO3)3]n- [n = 4 for uranyl(VI), 5 for uranyl(V)]. The bond distances of U-Oax, U-Oeq, U-C, and U-Odist are determined to be 1.81, 2.44, 2.92, and 4.17 A for the uranyl(VI) complex and 1.91, 2.50, 2.93, and 4.23 A for the uranyl(V) complex, respectively. The validity of the structural parameters obtained from EXAFS has been supported by quantum chemical calculations for the uranyl(VI) complex. The uranium LI- and LIII-edge X-ray absorption near-edge structure spectra have been interpreted in terms of electron transitions and multiple-scattering features. PMID- 17417837 TI - Intermolecular electron transfer from photogenerated Ru(bpy)3+ to [2Fe2S] model complexes of the iron-only hydrogenase active site. AB - Visible light-driven intermolecular electron transfer was observed from a reduced species Ru(bpy)3+, photogenerated via a reductive quenching of the ruthenium photosensitizer by a diethyldithiocarbamate anion, to bioinspired [2Fe2S] model complexes of the iron-only hydrogenase active site. The results indicate that Ru(bpy)32+ can act as a photoactive functional model of the [4Fe4S] cluster, playing the role of an electron-transfer relay. The photogenerated FeIFe0 species, which is proposed to be a crucial intermediate for proton reduction catalyzed electrochemically by the [2Fe2S] complexes, gives promise in the light driven dihydrogen evolution using diiron complexes as surrogates of noble platinum catalysts. PMID- 17417838 TI - Ultrafast proton transfer to solvent: molecularity and intermediates from solvation- and diffusion-controlled regimes. AB - Photoinduced proton transfer (PT) from cations 6-hydroxyquinolinium (6HQc) and 6 hydroxy-1-methylquinolinium (6MQc) to water and alcohols, and solvation of the zwitterionic conjugate base 1-methylquinolinium-6-olate (6MQz) were studied with stationary and transient absorption spectroscopy and by quantum chemical calculations. Transient emission spectra from 6MQz in acetonitrile and protic solvents shift dynamically to the red without changing their shape and intensity. The shift matches the solvation correlation function C(t) either measured with known solvatochromic probes coumarin 343 and coumarin 153 or derived from infrared/dielectric-loss data on neat solvents. This indicates that 6MQz monitors the solvation dynamics and that no intramolecular electron transfer occurs on a subpicosecond or longer time scale. The PT dynamics S(t) from 6HQc and 6MQc closely follows C(t), being initially 2-3 times slower. This allows for the conclusion that PT is controlled by solvation, with a barrier of 2 kJ/mol. In water, a pre-condition of this ultrafast reaction seems to be hydrogen-bonding between the negatively charged oxygen and two water molecules, resulting in a complex 6HQc:H2O:H2O. The complex is stable due to a high (47 kJ/mol) bonding energy between 6HQc and a water molecule. In acetonitrile, the reaction equilibrium is strongly shifted to the cation. There an intermediate PT state was detected, which may be ascribed to the cationic form 6HQc:H2O due to residual water impurities. In water-acetonitrile mixtures, the ultrafast solvent controlled PT is followed by a diffusion-controlled reaction; the measured rate kD approximately 1010 s-1 M-1 is characteristic for simple bimolecular diffusion. The dependence of the short-time PT signal on water concentration can be fitted with a Poisson distribution of water molecules around the cation. Altogether, the short-time and long-time behaviors provide strong evidence that diffusion of only one water molecule is sufficient to detach the proton. Subsequent solvent stabilization of the products completes the PT reaction. PMID- 17417839 TI - The origins of femtomolar protein-ligand binding: hydrogen-bond cooperativity and desolvation energetics in the biotin-(strept)avidin binding site. AB - The unusually strong reversible binding of biotin by avidin and streptavidin has been investigated by density functional and MP2 ab initio quantum mechanical methods. The solvation of biotin by water has also been studied through QM/MM/MC calculations. The ureido moiety of biotin in the bound state hydrogen bonds to five residues, three to the carbonyl oxygen and one for each--NH group. These five hydrogen bonds act cooperatively, leading to stabilization that is larger than the sum of individual hydrogen-bonding energies. The charged aspartate is the key residue that provides the driving force for cooperativity in the hydrogen bonding network for both avidin and streptavidin by greatly polarizing the urea of biotin. If the residue is removed, the network is disrupted, and the attenuation of the energetic contributions from the neighboring residues results in significant reduction of cooperative interactions. Aspartate is directly hydrogen-bonded with biotin in streptavidin and is one residue removed in avidin. The hydrogen-bonding groups in streptavidin are computed to give larger cooperative hydrogen-bonding effects than avidin. However, the net gain in electrostatic binding energy is predicted to favor the avidin-bicyclic urea complex due to the relatively large penalty for desolvation of the streptavidin binding site (specifically expulsion of bound water molecules). QM/MM/MC calculations involving biotin and the ureido moiety in aqueous solution, featuring PDDG/PM3, show that water interactions with the bicyclic urea are much weaker than (strept)avidin interactions due to relatively low polarization of the urea group in water. PMID- 17417840 TI - Spin-state selective carbon-detected HNCO with TROSY optimization in all dimensions and double echo-antiecho sensitivity enhancement in both indirect dimensions. AB - A carbon-detected TROSY-optimized experiment correlating 1HN, 15N, and 13C' resonances, referred to as c-TROSY-HNCO is presented, in which the 1HN and 15N TROSY effects are maintained in both indirect dimensions, while the directly detected 13C' is doubly TROSY-optimized with respect to 1HN and 15N. A new strategy for sensitivity enhancement, the so-called double echo-antiecho (dEA), is described and implemented in the c-TROSY-HNCO experiment. dEA offers sensitivity enhancement of square root of 2 in both indirect dimensions and is generally applicable to many multidimensional experiments. A carbon-detected HNCO experiment, c-HNCO, without TROSY optimization and sensitivity enhancement is also designed for comparison purposes. Relaxation simulations show that for a protein with a rotational correlation time of 10 ns or larger, the c-TROSY-HNCO experiment displays comparable or higher signal-to-noise (S/N) ratios than the c HNCO experiment, although the former selects only 1/4 of the initial magnetization relative to the later. The high resolution afforded in the directly detected carbon dimension allows direct measurement of the doublet splitting to extract 1JCalphaC' scalar and 1DCalphaC' residual dipolar couplings. Simulations indicate that the c-TROSY-HNCO experiment offers higher precision (lower uncertainty) compared to the c-HNCO experiment for larger proteins. The experiments are applied to 15N/13C/2H/[Leu,Val]-methyl-protonated IIBMannose, a protein of molecular mass 18.6 kDa with a correlation time of approximately 10 ns at 30 degrees C. The experimental pairwise root-mean-square deviation for the measured 1JCalphaC' couplings obtained from duplicate experiments is 0.77 Hz. By directly measuring the doublet splitting, the experiments described here are expected to be much more tolerant to nonuniform values of 1JCalphaC' (or 1JCalphaC' + 1DCalphaC' for aligned samples) and pulse imperfections due to the smaller number of applied pulses in the "out-and-stay" coherence transfer in the c-HNCO-TROSY experiment relative to conventional 1H-detected "out-and-back" quantitative J correlation experiments. A carbon-detected TROSY-optimized experiment correlating 1HN, 15N, and 13C' resonances, referred to as c-TROSY-HNCO is presented, in which the 1HN and 15N TROSY effects are maintained in both indirect dimensions, while the directly detected 13C' is doubly TROSY-optimized with respect to 1HN and 15N. A new strategy for sensitivity enhancement, the so called double echo-antiecho (dEA), is described and implemented in the c-TROSY HNCO experiment. dEA offers sensitivity enhancement of in both indirect dimensions and is generally applicable to many multidimensional experiments. PMID- 17417841 TI - Circular DNA and DNA/RNA hybrid molecules as scaffolds for ricin inhibitor design. AB - Ricin Toxin A-chain (RTA) catalyzes the hydrolytic depurination of A4324, the first adenosine of the GAGA tetra-loop portion of 28S eukaryotic ribosomal RNA. Truncated stem-loop versions of the 28S rRNA are RTA substrates. Here, we investigate circular DNA and DNA/RNA hybrid GAGA sequence oligonucleotides as minimal substrates and inhibitor scaffolds for RTA catalysis. Closing the 5'- and 3'-ends of a d(GAGA) tetraloop creates a substrate with 92-fold more activity with RTA (kcat/Km) than that for the d(GAGA) linear form. Circular substrates have catalytic rates (kcat) comparable to and exceeding those of RNA and DNA stem loop substrates, respectively. RTA inhibition into the nanomolar range has been achieved by introducing an N-benzyl-hydroxypyrrolidine (N-Bn) transition state analogue at the RTA depurination site in a circular GAGA motif. The RNA/DNA hybrid oligonucleotide cyclic GdAGA provides a new scaffold for RTA inhibitor design, and cyclic G(N-Bn)GA is the smallest tight-binding RTA inhibitor (Ki = 70 nM). The design of such molecules that lack the base-paired stem-loop architecture opens new chemical synthetic approaches to RTA inhibition. PMID- 17417842 TI - Intermolecular activation of C-X (X = H, O, F) bonds by a TiCt?Bu linkage. PMID- 17417843 TI - Processive rotaxane systems. Studies on the mechanism and control of the threading process. AB - The threading behavior of a zinc analogue of a previously reported processive manganese porphyrin catalyst onto a series of polymers of different lengths is reported. It is demonstrated that the speed of the threading process is determined by the opening of the cavity of the toroidal porphyrin host, which can be tuned with the help of axial ligands that coordinate to the metal center in the porphyrin. PMID- 17417844 TI - Environmental modulation of protein cation-pi interactions. PMID- 17417846 TI - Magnetic bistability and single-crystal-to-single-crystal transformation induced by guest desorption. PMID- 17417845 TI - Heterolytic activation of H-X (X = H, Si, B, and C) bonds: an experimental and theoretical investigation. AB - The highly electrophilic, coordinatively unsaturated, 16-electron [Ru(P(OH)3)(dppe)2][OTf]2 (dppe = Ph2PCH2CH2PPh2) complex 1 activates the H-H, the Si-H, and the B-H bonds, in H2(g), EtMe2SiH and Et3SiH, and H3B.L (L = PMe3, PPh3), respectively, in a heterolytic fashion. The heterolysis of H2 involves an eta2-H2 complex (observable at low temperatures), whereas the computations indicate that those of the Si-H and the B-H bonds proceed through unobserved eta1 species. The common ruthenium-containing product in these reactions is trans [Ru(H)(P(OH)3)(dppe)2][OTf], 2. The [Ru(P(OH)3)(dppe)2][OTf]2 complex is unique with regard to activating the H-H, the Si-H, and the B-H bonds in a heterolytic manner. These reactions and the heterolytic activation of the C-H bond in methane by the model complex [Ru(POH)3)(H2PCH2CH2PH2)2][Cl][OTf], 4, have been investigated using computational methods as well, at the B3LYP/LANL2DZ level. While the model complex activates the H-H, the Si-H, and the B-H bonds in H2, SiH4, and H3B.L (L = PMe3, PPh3), respectively, with a low barrier, activation of the C-H bond in CH4 involves a transition state of 57.5 kcal/mol high in energy. The inability of the ruthenium complex to activate CH4 is due to the undue stretching of the C-H bond needed at the transition state, in comparison to the other substrates. PMID- 17417847 TI - Polyfluorinated amino acids for sensitive 19F NMR-based screening and kinetic measurements. AB - Two novel series of polyfluorinated amino acids (PFAs) were designed and synthesized according to a very short and scalable synthetic sequence. The advantages and limitations of these moieties for screening purposes are presented and discussed. The potential applications of these PFAs were tested with their incorporation into small arginine-containing peptides that represent suitable substrates for the enzyme trypsin. The enzymatic reactions were monitored by 19F NMR spectroscopy, using the 3-FABS (three fluorine atoms for biochemical screening) technique. The high sensitivity achieved with these PFAs permits a reduction in substrate concentration required for 3-FABS. This is relevant in the utilization of 3-FABS in fragment-based screening for identification of small scaffolds that bind weakly to the receptor of interest. The large dispersion of 19F isotropic chemical shifts allows the simultaneous measurement of the efficiency of the different substrates, thus identifying the best substrate for screening purposes. Furthermore, the knowledge of KM and Kcat for the different substrates allows the identification of the structural motifs responsible for the binding affinity to the receptor and those affecting the chemical steps in enzymatic catalysis. This enables the construction of suitable pharmacophores that can be used for designing nonpeptidic inhibitors with high affinity for the enzyme or molecules that mimic the transition state. The novel PFAs can also find useful application in the FAXS (fluorine chemical shift anisotropy and exchange for screening) experiment, a 19F-based competition binding assay for the detection of molecules that inhibit the interaction between two proteins. PMID- 17417848 TI - Rh(I)-catalyzed carbonylative cyclization reactions of alkynes with 2 bromophenylboronic acids leading to indenones. AB - The Rh-catalyzed reaction of alkynes with 2-bromophenylboronic acids involves carbonylative cyclization to give indenones. The key steps in the reaction involve the addition of an arylrhodium(I) species to an alkyne and the oxidative addition of C-Br bonds on the adjacent phenyl ring to give vinylrhodium(I) species II. The regioselectivity depends on both the electronic and the steric nature of the substituents on the alkynes. A bulky group and an electron withdrawing group favor the -position of indenones. In the case of silyl- or ester-substituted alkynes, the regioselectivity is extremely high. The selectivity increases in the order SiMe3 > COOR >> aryl >> alkyl. The reaction of norbornene with 2-bromophenylboronic acids under 1 atm of CO gives the corresponding indanone derivative. The reaction of alkynes with 2 bromophenylboronic acids under nitrogen gives naphthalene derivatives, in which two molecules of alkynes are incorporated. A vinylrhodium complex similar to II can also be generated by a different route by employing 2 bromophenyl(trimethylsilyl)acetylene and arylboronic acids in the presence of Rh(I) complex as the catalyst, resulting in the formation of indenones. The reaction of 1-(2-bromophenyl)-hept-2-yn-1-one with PhB(OH)2 in the presence of Rh(I) complex also resulted in carbonylative cyclization to give an indan-1,3 dione derivative. PMID- 17417849 TI - Lamellar structure in poly(ala-gly) determined by solid-state NMR and statistical mechanical calculations. AB - Lamellar structure of poly(Ala-Gly) or (AG)n in the solid was examined using 13C solid-state NMR and statistical mechanical approaches. Two doubly labeled versions, [1-13C]Gly14[1-13C]Ala15- and [1-13C]Gly18[1-13C]Ala19 of (AG)15 were examined by two-dimensional (2D) 13C spin diffusion NMR in the solid state. In addition five doubly labeled [15N,13C]-versions of the same peptide, (AG) 15 and 15 versions labeled [3-13C] in each of the successive Ala residues were utilized for REDOR and 13C CP/MAS NMR measurements, respectively. The observed spin diffusion NMR spectra were consistent with a structure containing a combination of distorted beta-turns with a large distribution of the torsion angles and antiparallel beta-sheets. The relative proportion of the distorted beta-turn form was evaluated by examination of 13C CP/MAS NMR spectra of [3-13C]Ala-(AG)15. In addition, REDOR determinations showed five kinds of atomic distances between doubly labeled 13C and 15N nuclei which were also interpreted in terms of a combination of beta-sheets and beta-turns. Our statistical mechanical analysis is in excellent agreement with our Ala Cbeta 13C CP/MAS NMR data strongly suggesting that (AG)15 has a lamellar structure. PMID- 17417850 TI - Determining the orientation of uniaxially rotating membrane proteins using unoriented samples: a 2H, 13C, AND 15N solid-state NMR investigation of the dynamics and orientation of a transmembrane helical bundle. AB - Membrane protein orientation has traditionally been determined by NMR using mechanically or magnetically aligned samples. Here we show a new NMR approach that abolishes the need for preparing macroscopically aligned membranes. When the protein undergoes fast uniaxial rotation around the bilayer normal, the 0 degrees -frequency of the motionally averaged powder spectrum is identical to the frequency of the aligned protein whose alignment axis is along the magnetic field. Thus, one can use unoriented membranes to determine the orientation of the protein relative to the bilayer normal. We demonstrate this approach on the M2 transmembrane peptide (M2TMP) of influenza A virus, which is known to assemble into a proton-conducting tetrameric helical bundle. The fast uniaxial rotational diffusion of the M2TMP helical bundle around the membrane normal is characterized via 2H quadrupolar couplings, C-H and N-H dipolar couplings, 13C chemical shift anisotropies, and 1H T1rho relaxation times. We then show that 15N chemical shift anisotropy and N-H dipolar coupling measured on these powder samples can be analyzed to yield precise tilt angles and rotation angles of the helices. The data show that the tilt angle of the M2TMP helices depends on the membrane thickness to reduce the hydrophobic mismatch. Moreover, the orientation of a longer M2 peptide containing both the transmembrane domain and cytoplasmic residues is similar to the orientation of the transmembrane domain alone, suggesting that the transmembrane domain regulates the orientation of this protein and that structural information obtained from M2TMP may be extrapolated to the longer peptide. This powder-NMR approach for orientation determination is generally applicable and can be extended to larger membrane proteins. PMID- 17417851 TI - Tunable-alignment chiral system based on gelatin for NMR spectroscopy. PMID- 17417852 TI - Molecular basis for water-promoted supramolecular chirality inversion in helical rosette nanotubes. AB - Helical rosette nanotubes (RNTs) are obtained through the self-assembly of the GwedgeC motif, a self-complementary DNA base analogue featuring the complementary hydrogen bonding arrays of both guanine and cytosine. The first step of this process is the formation of a 6-membered supermacrocycle (rosette) maintained by 18 hydrogen bonds, which then self-organizes into a helical stack defining a supramolecular sextuple helix whose chirality and three-dimensional organization arise from the chirality, chemical structure, and conformational organization of the GwedgeC motif. Because a chiral GwedgeC motif is predisposed to express itself asymmetrically upon self-assembly, there is a natural tendency for it to form one chiral RNT over its mirror image. Here we describe the synthesis and characterization of a chiral GwedgeC motif that self-assembles into helical RNTs in methanol, but undergoes mirror image supramolecular chirality inversion upon the addition of very small amounts of water (<1% v/v). Extensive physical and computational studies established that the mirror-image RNTs obtained, referred to as chiromers, result from thermodynamic (in water) and kinetic (in methanol) self-assembly processes involving two conformational isomers of the parent GwedgeC motif. Although derived from conformational states, the chiromers are thermodynamically stable supramolecular species, they display dominant/recessive behavior, they memorize and amplify their chirality in an achiral environment, they change their chirality in response to solvent and temperature, and they catalytically transfer their chirality. On the basis of these studies, a detailed mechanism for supramolecular chirality inversion triggered by specific molecular interactions between water molecules and the GwedgeC motif is proposed. PMID- 17417853 TI - Thermal control over the topology of cleavable polymers: from linear to hyperbranched structures. PMID- 17417854 TI - Lipid transmembrane asymmetry and intrinsic membrane potential: two sides of the same coin. PMID- 17417855 TI - Kedarcidin chromophore: synthesis of its proposed structure and evidence for a stereochemical revision. PMID- 17417856 TI - Substrate profiling of protein tyrosine phosphatase PTP1B by screening a combinatorial peptide library. PMID- 17417857 TI - X-shaped rigid arylethynes to mediate the assembly of nanoparticles. PMID- 17417858 TI - Selective activation and functionalization of linear alkanes initiated under ambient conditions by a tungsten allyl nitrosyl complex. PMID- 17417861 TI - Long-distance radical cation hopping in DNA: the effect of thymine-Hg(II)-thymine base pairs. AB - Thymine-Hg(II)-thymine base pairs have been incorporated in an oligonucleotide duplex to study their effect on DNA-mediated charge transport. The introduction of a formally charged Hg atom inside the DNA base core does not significantly alter the charge hopping and trapping properties, as discussed in this paper. Hg(II) replaces the protons normally found on thymines within the complex and acts like a "big proton" in terms of its role in DNA charge transport. PMID- 17417862 TI - How accurate are DFT treatments of organic energies? AB - Increasing awareness that popular functionals fail to describe many energies accurately has ended expectations of black-box DFT usage. The performance of nine density functionals, compared by computing the bond separation energies of 72 illustrative hydrocarbons with available experimental data, reveals that only Zhao and Truhlar's recently proposed M05-2X functional, with a 2.13 kcal/mol average deviation from experiment, performs satisfactorily. B3LYP and other functionals show larger deviations. PMID- 17417863 TI - One-pot regioselective synthesis of chromanyl(phenyl)-lambda(3)-iodanes: tandem oxidative cyclization and lambda(3)-iodanation of 3-phenylpropanols. AB - Reaction of 3-phenylpropanol with an activated iodosylbenzene-18-crown-6 complex [PhI(OH)BF4-18C6] in dichloromethane in the presence of BF3-Et2O afforded directly the 6-chromanyl(phenyl)-lambda3-iodane-18C6 complex through tandem oxidative intramolecular cyclization yielding chroman and its regioselective phenyl-lambda3-iodanation. PMID- 17417864 TI - Physicochemical properties of native adzuki bean (Vigna angularis) 7S globulin and the molecular cloning of its cDNA isoforms. AB - 7S globulin (vicilin), the major seed storage protein in adzuki bean [Vigna angularis], was purified by ammonium sulfate fractionation, gel filtration column chromatography, and anion-exchange column chromatography that resulted in two fractions. On SDS-PAGE, both fractions gave two major and some minor bands, but there was a difference in the minor band compositions between the two fractions. Thermal stability, solubility, surface hydrophobicity, and emulsifying ability of these three samples were analyzed. Although there was no difference in solubility and emulsifying ability among the samples, thermal stability and surface hydrophobicity were different. These differences might be due to the differences in subunit compositions. cDNAs were cloned by reverse transcription-polymerase chain reaction (RT-PCR) using primers designed on the basis of the determined N terminal sequences of the major bands. We obtained three isoforms of cDNAs, which had highest homology with the mung bean 8Salpha globulin (7S globulin), and then soybean beta-conglycinin (7S globulin) beta subunit among legume plants. Adzuki bean 7S globulin isoforms contain more methionine and tryptophan than mung bean 8Salpha globulin and soybean beta-conglycinin beta subunit. In addition, high mannose types of glycans were attached to two or one N-glycosylation sites of adzuki bean 7S globulins. PMID- 17417865 TI - Role of kappa-casein in the association of denatured whey proteins with casein micelles in heated reconstituted skim milk. AB - Reconstituted skim milk at pH from 6.5 to 7.1 was unheated, preheated (68 degrees C/20 min), or heated at 90 degrees C for 20-30 min. On preheating, the size of the casein micelles decreased by about 5-20 nm, with a greater effect at higher pH. The casein micelle size of the heated milk at pH 6.5 increased by about 30 nm when compared to that of the unheated or preheated milk. As the pH was increased before heating, the particle size gradually decreased so that, at pH 7.1, the size was markedly smaller than that for the unheated milk and slightly smaller than that for the preheated milk. High levels (about 85%) of denatured whey protein associated with the casein micelles at pH 6.5, and this level decreased as the pH increased so that, at pH 7.1, low levels (about 15%) were associated with the micelles. Low levels of alphaS-casein and beta-casein were found in the serum regardless of the heat treatment or the pH of the milk. At pH 6.5, low levels (about 10%) of kappa-casein were also found in the milk serum. In the unheated milk, the level of serum kappa-casein increased slightly with increasing pH; in the heated samples, the level of serum kappa-casein increased markedly and linearly with increasing pH so that, at pH 7.1, about 70% of the kappa-casein was in the serum phase. The results of this study indicate that the pH dependence of the levels of serum phase kappa-casein may be responsible for the change in distribution of the whey proteins between the colloidal and serum phases. This is the first report to demonstrate significant levels of dissociation of kappa casein from the micelles at pH between 6.5 and 6.7, although this dissociation phenomenon is well known on heating milk at high temperatures at pH above 6.7. PMID- 17417866 TI - Freshness and quality criteria of iced farmed Senegalese sole (Solea senegalensis). AB - Senegalese sole (Solea senegalensis) is a high-value commercial species with increasing importance in aquaculture. The aim of this work was to study the quality changes of this species during chilled storage under refrigeration, through sensory and chemical methods. In particular, the optimization of a quality index method (QIM) scheme was proposed as well as the definition of sensory and chemical quality criteria. A shelf life of 15 days was reported, and a QIM scheme based on a total of 22 demerit points (dp) was proposed. Biogenic amines were never detected, and the usual spoilage indicators, such as TVB-N and TMA-N, were not significantly produced during the period of sensory-acceptable quality (15 days). On the basis of the significant correlations (p<0.001) between sensory data and Ki values, a quality index (QI) 19 dp and a Ki>40% indicate unacceptable quality of iced Senegalese sole. PMID- 17417867 TI - Nucleophilic reactions of phorate and terbufos with reduced sulfur species under anoxic conditions. AB - The reactions of phorate and terbufos with bisulfide (HS-), polysulfide (Sn2-), thiosulfate (S2O32-), and thiophenolate (PhS-) were examined in well-defined aqueous solution under anoxic conditions to investigate their role in the degradations of phorate and terbufos. Reactions were monitored at various concentrations of reduced sulfur species to obtain the second-order rate constants. The reactivity of the reduced sulfur species decreased in the order Sn2- > PhS- > HS- > S2O32-. Hydrolysis products, formaldehyde and diethyl disulfide/di-tert-butyl disulfide, indicated that OH-/H2O attacked the carbon atom between the two sulfur atoms, the so-called thioacetal carbon, which is very reactive due to the presence of the two neighboring sulfur atoms. The reaction of phorate and terbufos with PhS- was investigated to study the transformation products in the reactions with reduced sulfur species. The transformation products demonstrated that the observed increase in rate constants in the reaction with reduced sulfur species compared to hydrolysis could result from the nucleophilic attack of reduced sulfur species at the alpha-carbon of the ethoxy group and at the thioacetal carbon atom. The temperature dependence of measured second-order rate constants of the reaction of phorate and terbufos with HS- over 25-50 degrees C was investigated to explore activation parameters, which are not significantly different for phorate and terbufos. All of the observations may imply similar pathways in the degradation of phorate and terbufos in the presence of reduced sulfur species. Slightly higher hydrolysis rates of terbufos and second-order reaction rate constants for the reactions with sulfur species of terbufos compared with those for phorate are observed, which could be attributed to the slightly different substituents. PMID- 17417868 TI - Fish species identification in surimi-based products. AB - Whole fish morphologically identified as belonging to Theragra chalcogramma, Merluccius merluccius, Merluccius hubbsi, and Merluccius capensis and 19 fish products commercialized as surimi with different commercial brands and labeled as T. chalcogramma were analyzed by direct sequence analysis of the cytochrome b gene. A phylogenetic analysis of surimi products was performed as well. Results demonstrated that mislabeling is a large-scale phenomenon, since 84.2% of surimi based fish products sold as T. chalcogramma (16/19) were prepared with species different from the one declared. In fact, only three samples (samples 15-17) were found to belong to T. chalcogramma. In the remaining samples, Merluccidae (samples 4-14), Gadidae (samples 18 and 19), Sparidae (sample 1), and Pomacentridae (samples 2 and 3) families were detected. A phylogenetic tree was constructed, and the bootstrap value was calculated. According to this methodology, 11 samples were grouped in the same clade as Merluccius spp. PMID- 17417869 TI - Racemization at C-2 of naringin in sour oranges with increasing maturity determined by chiral high-performance liquid chromatography. AB - Naringin is the major flavanone-7-O-glycoside of sour orange, and it is mainly responsible for the bitter taste of the fruit. The relative content of (2S)- and (2R)-naringin in the albedo of sour oranges during maturation in the entire season was determined by normal phase HPLC using Chiralcel OD-H as chiral stationary phase and n-hexane/ethanol doped with 0.5% of TFA as mobile phase. HPLC traces were complicated by the presence of (2S)-neohesperidin, and a software-guided analysis was developed to assess the relative amount of the C-2 diastereomers of naringin. A sigmoid curve was obtained showing variation from 94% of (2S)-naringin in very immature fruits to 69.7% in mature fruit samples. Spontaneous epimerization of (2S)-naringin in the ethanolic solutions of the albedo was noted after prolonged keeping for less acid samples. The separation of the C-2 diastereomers of other flavanoid-7-O-glycosides present in Citrus (neoeriocitrin, neohesperidin, and eriocitrin) was also obtained by chiral HPLC and afforded the identification, including stereochemistry, of the eluting peaks in the HPLC traces of the albedo extracts. PMID- 17417870 TI - Oral toxicity of beta-N-acetyl hexosaminidase to insects. AB - Insect chitin is a potential target for resistance plant proteins, but plant derived chitin-degrading enzymes active against insects are virtually unknown. Commercial beta-N-acetylhexosaminidase (NAHA), a chitin-degrading enzyme from jack bean Canavalia ensiformis, caused significant mortality of fall armyworm Spodoptera frugiperda larvae at 75 microg/gm, but no significant mortality was noted with Aspergillus niger NAHA. Maize Zea mays callus transformed to express an Arabidopsis thaliana clone that putatively codes for NAHA caused significantly higher mortality of cigarette beetle Lasioderma serricorne larvae and significantly reduced growth rates (as reflected by survivor weights) of S. frugiperda as compared to callus that expressed control cDNAs. Tassels from model line Hi-II maize (Z. mays) plants transformed with the NAHA gene fed to S. frugiperda caused significantly higher mortality than tassels transformed to express glucuronidase; mortality was significantly correlated with NAHA expression levels detected histochemically. Leaf disks from inbred Oh43 maize plants transformed with the NAHA gene on average had significantly less feeding by caterpillars than null transformants. Leaf disks of Oh43 transformants caused significant mortality of both S. frugiperda and corn earworm Helicoverpa zea larvae, which was associated with higher expression levels of NAHA detected by isoelectric focusing, histochemically, or with antibody. Overall, these results suggest that plant NAHA has a role in insect resistance. Introduction of NAHA genes or enhancement of activity through breeding or genetic engineering has the potential to significantly reduce insect damage and thereby indirectly reduce mycotoxins that are harmful to animals and people. PMID- 17417871 TI - Glyphosate-resistant and -susceptible soybean (Glycine max) and canola (Brassica napus) dose response and metabolism relationships with glyphosate. AB - Experiments were conducted to determine (1) dose response of glyphosate-resistant (GR) and -susceptible (non-GR) soybean [Glycine max (L.) Merr.] and canola (Brassica napus L.) to glyphosate, (2) if differential metabolism of glyphosate to aminomethyl phosphonic acid (AMPA) is the underlying mechanism for differential resistance to glyphosate among GR soybean varieties, and (3) the extent of metabolism of glyphosate to AMPA in GR canola and to correlate metabolism to injury from AMPA. GR50 (glyphosate dose required to cause a 50% reduction in plant dry weight) values for GR (Asgrow 4603RR) and non-GR (HBKC 5025) soybean were 22.8 kg ae ha-1 and 0.47 kg ha-1, respectively, with GR soybean exhibiting a 49-fold level of resistance to glyphosate as compared to non GR soybean. Differential reduction in chlorophyll by glyphosate was observed between GR soybean varieties, but there were no differences in shoot fresh weight reduction. No significant differences were found between GR varieties in metabolism of glyphosate to AMPA, and in shikimate levels. These results indicate that GR soybean varieties were able to outgrow the initial injury from glyphosate, which was previously caused at least in part by AMPA. GR50 values for GR (Hyola 514RR) and non-GR (Hyola 440) canola were 14.1 and 0.30 kg ha-1, respectively, with GR canola exhibiting a 47-fold level of resistance to glyphosate when compared to non-GR canola. Glyphosate did not cause reduction in chlorophyll content and shoot fresh weight in GR canola, unlike GR soybean. Less glyphosate (per unit leaf weight) was recovered in glyphosate-treated GR canola as compared to glyphosate-treated GR soybean. External application of AMPA caused similar injury in both GR and non-GR canola. The presence of a bacterial glyphosate oxidoreductase gene in GR canola contributes to breakdown of glyphosate to AMPA. However, the AMPA from glyphosate breakdown could have been metabolized to nonphytotoxic metabolites before causing injury to GR canola. Injury in GR and non-GR canola from exogenous application of AMPA was similar. PMID- 17417872 TI - Representation, comparison, and interpretation of metabolome fingerprint data for total composition analysis and quality trait investigation in potato cultivars. AB - Understanding attributes of crop varieties and food raw materials underlying desirable characteristics is a significant challenge. Metabolomics technology based on flow infusion electrospray ionization mass spectrometry (FIE-MS) has been used to investigate the chemical composition of potato cultivars associated with quality traits in harvested tubers. Through the combination of metabolite fingerprinting with random forest data modeling, a subset of metabolome signals explanatory of compositional differences between individual genotypes were ranked for importance. Interpretative analysis of highlighted signals based on ranking behavior, intensity correlations, and mathematical relationships of ion masses correctly predicted metabolites associated with flavor and pigmentation traits in potato tubers. GC-MS profiling was used to further validate proposed compositional differences. The potential for the development of a database strategy for large scale, long-term projects requiring comparison of chemical composition in plant breeding, mutant population analysis in functional genomics experiments, or food raw material analysis is described. PMID- 17417873 TI - Solid-state nuclear magnetic resonance measurements of HIV fusion peptide to lipid distances reveal the intimate contact of beta strand peptide with membranes and the proximity of the Ala-14-Gly-16 region with lipid headgroups. AB - Human immunodeficiency virus (HIV) infection begins with fusion between viral and host cell membranes and is catalyzed by the HIV gp41 fusion protein. The approximately 20 N-terminal apolar residues of gp41 are called the HIV fusion peptide (HFP), interact with the host cell membrane, and play a key role in fusion. In this study, the membrane location of peptides which contained the HFP sequence (AVGIGALFLGFLGAAGSTMGARS) was probed in samples containing either only phospholipids or phospholipids and cholesterol. Four HFPs were examined which each contained 13CO labeling at three sequential residues between G5 and G16. The 13CO chemical shifts indicated that HFP had predominant beta strand conformation over the labeled residues in the samples. The internuclear distances between the HFP 13CO groups and the lipid 31P atoms were measured using solid-state nuclear magnetic resonance rotational-echo double-resonance experiments. The shortest 13CO-31P distances of 5-6 A were observed for HFP labeled between A14 and G16 and correlated with intimate association of beta strand HFP and membranes. These results were confirmed with measurements using HFPs singly labeled with 13CO at A6 or A14. To our knowledge, these data are the first measurements of distances between HIV fusion peptide nuclei and lipid P, and qualitative models of the membrane location of oligomeric beta strand HFP which are consistent with the experimental data are presented. Observation of intimate contact between beta strand HFP and membranes provides a rationale for further investigation of the relationship between structure and fusion activity for this conformation. PMID- 17417874 TI - The autolytic regulation of human kallikrein-related peptidase 6. AB - Human kallikrein-related peptidase 6 (KLK6) is a member of the kallikrein family of serine-type proteases, characterized as an arginine-specific digestive-type protease capable of degrading a wide-variety of extracellular matrix proteins. KLK6 has been proposed to be a useful biomarker for breast and ovarian cancer prognosis, is abundantly expressed in the CNS and cerebrospinal fluid, and is intimately associated with regions of active inflammatory demyelination in multiple sclerosis (MS) lesions. Inhibition of KLK6 results in delayed onset and reduced severity of symptoms associated with experimental autoimmune encephalomyelitis, suggesting a key effector role for this protease in CNS inflammatory disease. KLK6 has been shown to autolytically cleave internally, leading to inactivation and suggesting a negative feedback inhibition control mechanism. Alternatively, the ability of KLK6 to self-activate has also been reported, suggesting a positive feedback activation loop control mechanism. Activation of pro-KLK6 requires hydrolysis after a Lys residue; however, KLK6 exhibits 2 order of magnitude reduced affinity for hydrolysis after Lys versus Arg residues; therefore, the ability to autolytically activate has been called into question. In the present study the catalytic activity of KLK6 toward its pro sequence and internal autolytic sequence is characterized. The results show that the ability of KLK6 to activate pro-KLK6 is essentially negligible when compared to the rate of the internal autolytic inactivation or to the ability of other proteases to activate pro-KLK6. The results thus show that the primary autolytic regulatory mechanism of KLK6 is negative feedback inhibition, and activation is likely achieved through the action of a separate protease. PMID- 17417875 TI - A conserved region between the heptad repeats of paramyxovirus fusion proteins is critical for proper F protein folding. AB - Paramyxoviruses are a diverse family that utilizes a fusion (F) protein to enter cells via fusion of the viral lipid bilayer with a target cell membrane. Although certain regions of the F protein are known to play critical roles in membrane fusion, the function of much of the protein remains unclear. Sequence alignment of a set of paramyxovirus F proteins and analysis utilizing Block Maker identified a region of conserved amino acid sequence in a large domain between the heptad repeats of F1, designated CBF1. We employed site-directed mutagenesis to analyze the function of completely conserved residues of CBF1 in both the simian virus 5 (SV5) and Hendra virus F proteins. The majority of CBF1 point mutants were deficient in homotrimer formation, proteolytic processing, and transport to the cell surface. For some SV5 F mutants, proteolytic cleavage and surface expression could be restored by expression at 30 degrees C, and varying levels of fusion promotion were observed at this temperature. In addition, the mutant SV5 F V402A displayed a hyperfusogenic phenotype at both 30 and 37 degrees C, indicating that this mutation allows for efficient fusion with only an extremely small amount of cleaved, active protein. The recently published prefusogenic structure of PIV5/SV5 F (Yin, H. S., et al. (2006) Nature 439, 38 44) indicates that residues within and flanking CBF1 interact with the fusion peptide domain. Together, these data suggest that CBF1-fusion peptide interactions are critical for the initial folding of paramyxovirus F proteins from this important viral family and can also modulate subsequent membrane fusion promotion. PMID- 17417876 TI - A single nucleotide linked to a switch in metal ion reactivity preference in the HDV ribozymes. AB - The two ribozymes of hepatitis delta virus (HDV) cleave faster in divalent metal ions than in monovalent cations, and a variety of divalent metal ions can act as catalysts in supporting these higher rates. Although the ribozymes are closely related in sequence and structure, they display a different metal ion preference; the genomic form cleaves moderately faster in Mg2+ than in Ca2+ while the reverse is true for the antigenomic ribozyme. This difference raises questions about understanding the catalytic role of the metal ion in the reaction. We found that the metal ion reactivity preference correlated with the identity of a single nucleotide 5' of the cleavage site (-1 position). It is a U in the genomic sequence and a C in the antigenomic sequence. With both ribozymes, the reactivity preference for Mg2+ and Ca2+ could be reversed with a change at this position (C to U or U to C). Moreover, with an A at position -1, there was a relative increase in cleavage rates in low concentrations of Mn2+ for both ribozymes. Metal ion reactivity preference was also linked to changes in pH, and the pH-rate profiles could be shifted with nucleotide changes at position -1. Together, the data provide biochemical evidence in support of an organized active site, as seen in the crystal structures, where at least one metal ion, an ionizable group, and the conformation of the phosphate backbone at the cleavage site interact in concert to promote cleavage. PMID- 17417877 TI - Reduction of high-affinity beta2-adrenergic receptor binding by hyperforin and hyperoside on rat C6 glioblastoma cells measured by fluorescence correlation spectroscopy. AB - Beta-adrenergic receptors (beta-AR) are potential targets for antidepressants. Desensitization and downregulation of beta-AR are discussed as possible modes of action for antidepressants. We have investigated the effects of hyperforin and hyperoside, compounds with potentially antidepressant activity from St. John's Wort, on the binding behavior and dynamics of beta2-AR in living rat C6 glioblastoma cells, compared to desipramine (desmethylimipramine; DMI) by means of fluorescence correlation spectroscopy (FCS) and fluorescence microscopy. FCS binding studies with the fluorescently labeled ligand Alexa532-noradrenaline (Alexa532-NA) binding to beta2-AR of C6 cells showed a significant reduction in total beta2-AR binding after preincubation with hyperforin and hyperoside for 3 days, respectively, which was also found for DMI. This was mainly observed in high-affinity receptor-ligand complexes with hindered lateral mobility (D2 = 1.1 (+/-0.4) microm2/s) in the biomembrane. However, internalization of beta2-AR was found neither in z-scans of these C6 cells nor in HEK 293 cells stably transfected with GFP-tagged beta2-adrenergic receptors (beta2AR-GFP) after incubation up to 6 days with either DMI, hyperforin, or hyperoside. Thus, under these conditions reduction of beta2-AR binding was not mediated by receptor internalization. Additionally, preincubation of C6 cells with DMI, hyperforin, and hyperoside led to a loss of second messenger cAMP after beta2-adrenergic stimulating conditions with terbutaline. Our current results indicate that hyperforin and hyperoside from St. John's Wort, as well as DMI, reduce beta2 adrenergic sensitivity in C6 cells, emphasizing the potential usefulness of St. John's Wort dry extracts in clinical treatment of depressive symptoms. PMID- 17417878 TI - Consolidation of the thioredoxin fold by peptide recognition: interaction between E. coli thioredoxin fragments 1-93 and 94-108. AB - Escherichia coli thioredoxin (TRX) catalyzes redox reactions via the reversible oxidation of the conserved active center WCGPC. TRX is a monomeric alpha/beta protein with a fold characterized by a central beta-sheet surrounded by alpha helical elements. The interaction of the C-terminal alpha-helix (helix 5) of TRX against the remainder of the protein involves the close packing of hydrophobic surfaces, opening the possibility of studying a fine-tuned molecular recognition phenomenon. To evaluate the relevance of this interaction on the folding mechanism of TRX, we characterize TRX1-93, a truncated variant of TRX devoid of the last stretch of 15 amino acid residues that includes helix 5. TRX1-93 may possibly represent a molecular form where the folding process becomes interrupted, giving rise to a structure exhibiting the features of a molten globule state. This was assessed by circular dichroism, intrinsic fluorescence, binding of the probe ANS, size-exclusion chromatography, limited proteolysis, and calorimetry. Remarkably, fragment TRX1-93 interacts with peptide TRX94-108 (KD approximately 2-12 microM), bringing forth the restoration of native-like signatures and enzymic function. This represents a molecular event of reciprocal structure selection where both partners gain order, thus leading to long-range consequences on conformation. In this context, the binding of the C-terminal helix could signify a late event in the consolidation of the overall TRX fold. PMID- 17417879 TI - Critical role of Arg/Lys343 in the species-dependent recognition of phosphatidylinositol by pulmonary surfactant protein D. AB - Surfactant protein D (SP-D) plays important roles in lung host defense. However, it can also recognize specific host molecules and contributes to surfactant homeostasis. The major known surfactant-associated ligand is phosphatidylinositol (PI). Trimeric neck-carbohydrate recognition domains (NCRDs) of rat and human SP D exhibited dose-dependent, calcium-dependent, and inositol-sensitive binding to solid-phase PI and to multilamellar PI liposomes. However, the rat protein exhibited a >5-fold higher affinity for solid-phase PI than the human NCRD. In addition, human dodecamers, but not full-length human trimers, efficiently coprecipitated with multilamellar PI liposomes in the presence of calcium. A human NCRD mutant resembling the rat and mouse proteins at position 343 (hR343K) showed much stronger binding to PI. A reciprocal rat mutant with arginine at the position of lysine 343 (rK343R) showed weak binding to PI, even weaker than that of the wild-type human protein. Crystal complexes of the human trimeric NCRD with myoinositol and inositol 1-phosphate showed binding of the equatorial OH groups of the cyclitol ring of the inositol to calcium at the carbohydrate binding site. Myoinositol binding occurred in two major orientations, while inositol 1 phosphate appeared primarily constrained to a single, different orientation. Our studies directly implicate the CRD in PI binding and reveal unexpected species differences in PI recognition that can be largely attributed to the side chain of residue 343. In addition, the studies indicate that oligomerization of trimeric subunits is an important determinant of recognition of PI by human SP-D. PMID- 17417880 TI - Mechanistic characterization of the bifunctional aminoglycoside-modifying enzyme AAC(3)-Ib/AAC(6')-Ib' from Pseudomonas aeruginosa. AB - A recently discovered bifunctional antibiotic-resistance enzyme named AAC(3) Ib/AAC(6')-Ib', from Pseudomonas aeruginosa, catalyzes acetylation of aminoglycoside antibiotics. Since both domains are acetyltransferases, each was cloned and purified for mechanistic studies. The AAC(3)-Ib domain appears to be highly specific to fortimicin A and gentamicin as substrates, while the AAC(6') Ib' domain exhibits a broad substrate spectrum. Initial velocity patterns indicate that both domains follow a sequential kinetic mechanism. The use of dead end and product inhibition and solvent-isotope effect reveals that both domains catalyze their reactions by a steady-state ordered Bi-Bi kinetic mechanism, in which acetyl-CoA is the first substrate that binds to the active site, followed by binding of the aminoglycoside antibiotic. Subsequent to the transfer of the acetyl group, acetylated aminoglycoside is released prior to coenzyme A. The merger of two genes to create a bifunctional enzyme with expanded substrate profile would appear to be a recent trend in evolution of resistance to aminoglycoside antibiotics, of which four examples have been documented in the past few years. PMID- 17417881 TI - Functional characterization of the heterooligomeric EbrAB multidrug efflux transporter of Bacillus subtilis. AB - The Bacillus subtilis genome contains two tandem genes, ebrA and ebrB, which encode two homologues of the SMR family of multidrug efflux transporters. The sequences of EbrA and EbrB are highly similar to each other and to that of EmrE, the prototypical SMR transporter of Escherichia coli. Drug resistance profiling and drug binding experiments showed that the presence of both EbrA and EbrB is required for proper transport function. EbrA and EbrB directly interact and combine to form a functional transporter. They likely form a heterodimer in analogy to the EmrE homodimer. Mutagenesis experiments indicate that the conserved membrane-embedded glutamates in the first transmembrane helices of both EbrA and EbrB are required for multidrug efflux activity. However, the two glutamates are nonequivalent since EbrA E15 is required for substrate binding while EbrB E14 is not. Our studies support a model in which functional residues in EbrAB are relegated to at least two sets that participate in distinct steps of the active drug transport process. PMID- 17417882 TI - The effect of aggregation on the electrical conductivity of spin-coated polymer/carbon nanotube composite films. AB - This paper considers the feasibility of replacing indium tin oxide (ITO) with spin-coated, polymer-based composite films that are filled with multiwalled carbon nanotubes (MWNTs). The coating mixture consists of a solvent with low volatility, a dissolved thermoplastic polymer, and MWNTs. The high aspect ratio of MWNTs and their good electrical conductivity enable electrical percolation at very low concentrations, so that films can be prepared that conduct electricity while retaining good optical transparency. Although the MWNTs are driven to aggregate by Van der Waals interactions, the high viscosity of the polymer/solvent solution enables the preparation of metastable, homogeneous dispersions. However, exposing the mixtures to shear leads to aggregation, the magnitude of which depends on the duration of the shear. This effect could be observed directly in spin-coated films using both optical microscopy and conductivity measurements, with aggregation causing a drop in conductivity at high nanotube loading, and more complex non-monotonic behavior at concentrations approaching the percolation threshold. PMID- 17417883 TI - Surface ionization state and nanoscale chemical composition of UV-irradiated poly(dimethylsiloxane) probed by chemical force microscopy, force titration, and electrokinetic measurements. AB - The surface chemistry and ionization state of cross-linked poly(dimethylsiloxane) (PDMS) exposed to UV/ozone were studied as a function of treatment time. Various complementary and independent experimental techniques were utilized, which yielded information on the macroscopic as well as the nanometric scale. The average chemical composition of the PDMS surface was quantitatively investigated by time-of-flight secondary ion mass spectrometry (ToF-SIMS). It was found that the top 1-2 nm surface layer was dominated by silanol groups (-SiOH) for which the concentration increased with increasing treatment dose. The lateral distributions of the silanol groups were analyzed on the nanometer scale by means of atomic force microscopy (AFM) with chemically functionalized tip probes in aqueous buffer solutions at varying pHs. Spatially dependent pull-off force curves (also called "force volume" imaging) indicated the presence of strong chemical heterogeneity of the probed surface. This heterogeneity took the form of patches of silanol functionalities with high local concentration surrounded by a matrix of predominantly hydrophobic domains at low pH. The average pull-off forces for the entire surface scanned were significantly reduced for pH values larger than a characteristic pK(a) constant (in the range between 4.5 and 5.5). The extent of the decrease in the pull-off force and the particular value of pK(a) were found to be a function of treatment time and to differ from the commonly reported values for silanol functional groups on a homogeneous silica surface. These dependences were ascribed to the evoking of a protonation/deprotonation process of the surface silanol groups which was sensitive to the hydrophobic/hydrophilic balance of their close molecular environment. Intermolecular hydrogen bonding may also account for the shifts in the surface pK(a). Furthermore, depending on the nature of the electrolyte, a third effect related to double layer composition, as determined by specific ion adsorption, was quantitatively analyzed by streaming potential measurements in the presence of sodium chloride and phosphate electrolytes. PMID- 17417884 TI - Simple correlation-corrected theory of systems described by screened coulomb interactions. AB - We present a simple correlation-corrected density functional treatment of dispersions containing macroions, where these are assumed to interact via screened Coulomb potentials, as given by Debye-Huckel theory. A straightforward mean-field description even fails to qualitatively capture important correlation effects displayed by such systems. However, if an effective, correlation corrected potential is adopted at short range, then the predictions are in qualitative and semiquantitative agreement with simulated results. The correlation corrections are evaluated in a manner that is completely analogous to those recently presented in correlation-corrected Poisson-Boltzmann theory (Forsman, J. J. Phys. Chem. B 2004, 108, 9236). The accuracy of the theory is evaluated by comparison with simulation data on systems displaying correlation generated packing effects and stratification forces. PMID- 17417885 TI - Preparation of size-controlled, highly populated, stable, and nearly monodispersed Ag nanoparticles in an organic medium from a simple interfacial redox process using a conducting polymer. AB - The reducing property of an organically soluble conducting polymer (poly(o methoxyaniline), POMA) is used to prepare monodisperse, size-controlled, highly populated, and highly stable silver nanoparticles in an organic medium through an interfacial redox process with an aqueous AgNO3 solution. The transition of emeraldine base (EB) to the pernigraniline base (PB) form of POMA occurs during nanoparticle formation, and the nitrogen atoms of POMA(PB) stabilize Ag nanoparticles by coordination to the adsorbed Ag(+) on the nanoparticle surface. The conductivity of the nanocomposite is on the order of 10(-11) S/cm, indicating that no doping of POMA occurs under the preparation conditions. The nanoparticles are free of excess oxidant and external stabilizer particles. The POMA (EB) concentration tailors the size of nanoparticles, and at its higher concentration (0.01% POMA with 0.01 N AgNO3), very dense Ag nanoparticles (6 x 10(15) particles/m(2)) of almost uniform size and shape are produced. The rate constant and Avrami exponent values of the nanoparticle formation are measured from the time-dependent UV-vis spectra using the Avrami equation. The Avrami exponent (n) values are close to 1, indicating 2D athermal nucleation with the circular shape of the nuclei having diffusion-controlled growth. The rate constant values are almost independent of AgNO3 concentration but are strongly dependent on POMA concentration. The higher rate constant with increasing POMA(EB) concentration has been attributed for the lowering of nanoparticle size due to increased nucleation density. PMID- 17417886 TI - Localization of sphingomyelin in cholesterol domains by imaging mass spectrometry. AB - The location of each lipid in a palmitoyloleoylphosphatidylcholine/18:0 sphingomyelin/cholesterol monolayer system is laterally resolved using imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) without the necessity of adding fluorescent labels. This system of coexisting immiscible liquid phases shows cholesterol domains with sizes and shapes comparable to those in the fluorescence microscopy literature. The results show that SM localizes with cholesterol and that palmitoyloleoylphosphatidylcholine is excluded. Moreover, the segregation is not complete, and there is a small amount of both phospholipids distributed throughout. PMID- 17417887 TI - Preparation and surface characterization of polymer nanoparticles designed for incorporation into hybrid materials. AB - We prepared water dispersions of poly(n-butyl methacrylate-st-butyl acrylate) crosslinked core-shell nanoparticles functionalized with different amounts of trimethoxisilane (TMS) groups in the outer shell. The purpose of the TMS groups is to chemically bind the rubbery particles to a nanostructured silica network, using sol-gel copolymerization. Here, we present nanoparticles containing 13 mol % and 30 mol % of TMS groups in the outer shell and compare their surface morphology with particles that do not contain TMS. The particles are prepared by a two-step seeded emulsion polymerization technique at neutral pH. In the first step, we obtained crosslinked seed particles (44 nm in diameter) by a batch process. In the second step, we used a semi-continuous emulsion polymerization technique under starved feed conditions to obtain monodispersed particles of controlled composition and size (ca. 100 nm in diameter). Fluorescence decay measurements were performed in situ on the dispersions, using a pair of cationic dyes adsorbed onto the surface of the nanoparticles: rhodamine 6G as the energy transfer donor and malachite green carbinol hydrochloride as the acceptor. The kinetics of Forster resonance energy transfer (FRET) between the dyes is sensitive to the donor-acceptor distance, allowing us to obtain the binding distribution of the dyes at the nanoparticle surface. For the unmodified nanoparticles, we found a dye distribution that corresponds to an average interface thickness of delta = (5.2 +/- 0.2) nm. For the samples containing 13 mol % and 30 mol % of TMS groups in the outer shell we obtained broader interfaces, with widths of delta = (6.2 +/- 0.2) nm and delta = (6.5 +/- 0.1) nm respectively. This broadening of the distribution with the surface modification is interpreted in terms of the increase in free volume of the shell caused by the TMS groups. Finally, we studied the effect of temperature on the water-polymer interface fuzziness, in order to evaluate the accessibility of the TMS groups during the sol-gel synthesis of nanostructured hybrid materials. PMID- 17417888 TI - Clathrate formation mechanism of supercritical hydrogen adsorption on copper(II) benzoate pyrazine. AB - The adsorption isotherms of supercritical hydrogen on [Cu2(bz)4(pyz)]n were measured at 77 K up to 10 MPa. The amount of supercritical hydrogen adsorbed on [Cu2(bz)4(pyz)]n at 77 K was 1.4 wt % at 10 MPa. The adsorption isotherms of supercritical hydrogen on [Cu2(bz)4(pyz)]n showed a stepwise adsorption that suggests clathrate formation between [Cu2(bz)4(pyz)]n and hydrogen molecules. PMID- 17417889 TI - Hard versus soft particle electrokinetics of silica colloids. AB - To verify the existence of a gel layer at the surface of silica, dependences of the electrophoretic mobility of fresh and aged colloidal silica particles on the KCl concentration are measured. These dependences, corrected for the relaxation/polarization effect, are fitted by analytical expressions based on the model of hard, soft, and brush surfaces. A bad fit is obtained for both silicas when its surface is considered ideal (hard). Much better fits are achieved with the invariable soft layer model for the fresh silica but especially for the aged silica whose surface is less charged probably as a result of an extension and/or loosening of the layer. A perfect fit is found for aged silica when applying a trivial model of the soft polyelectrolyte layer combined with the scaling model of polyelectrolyte brushes. PMID- 17417890 TI - Influence of Brownian motion on blood platelet flow behavior and adhesive dynamics near a planar wall. AB - We used the platelet adhesive dynamics computational method to study the influence of Brownian motion of a platelet on its flow characteristics near a surface in the creeping flow regime. Two important characterizations were done in this regard: (1) quantification of the platelet's ability to contact the surface by virtue of the Brownian forces and torques acting on it, and (2) determination of the relative importance of Brownian motion in promoting surface encounters in the presence of shear flow. We determined the Peclet number for a platelet undergoing Brownian motion in shear flow, which could be expressed as a simple linear function of height of the platelet centroid, H from the surface Pe (platelet) = . (1.56H + 0.66) for H > 0.3 microm. Our results demonstrate that at timescales relevant to shear flow in blood Brownian motion plays an insignificant role in influencing platelet motion or creating further opportunities for platelet-surface contact. The platelet Peclet number at shear rates >100 s-1 is large enough (>200) to neglect platelet Brownian motion in computational modeling of flow in arteries and arterioles for most practical purposes even at very close distances from the surface. We also conducted adhesive dynamics simulations to determine the effects of platelet Brownian motion on GPIbalpha-vWF-A1 single-bond dissociation dynamics. Brownian motion was found to have little effect on bond lifetime and caused minimal bond stressing as bond rupture forces were calculated to be less than 0.005 pN. We conclude from our results that, for the case of platelet-shaped cells, Brownian motion is not expected to play an important role in influencing flow characteristics, platelet-surface contact frequency, and dissociative binding phenomena under flow at physiological shear rates (>50 s( 1)). PMID- 17417891 TI - Interaction of metal porphyrins with fullerene C60: a new insight. AB - The electronic structure and bonding in the noncovalent, supramolecular complexes of fullerene C60 with a series of first-row transition metal porphines MP (M=Fe, Co, Ni, Cu, Zn) have been re-examined with DFT methods. A dispersion correction was made for the C60-MP binding energy through an empirical method (J. Comput. Chem. 2004, 25, 1463). Several density functionals and two types of basis sets were employed in the calculations. Our calculated results are rather different from those obtained in a recent paper (J. Phys. Chem. A 2005, 109, 3704). The ground state of C60.FeP is predicted to be high spin (S=2); the low-spin (S=0), closed-shell state is even higher in energy than the intermediate-spin (S=1) state. With only one electron in the Co-dz2 orbital, the calculated Co-C60 distance is in fact rather short, about 0.1 A longer than the Fe-C60 distance in high-spin C60.FeP. Double occupation of an M-dz2 orbital in MP prevents close association of any axial ligand, and so the Ni-C60, Cu-C60, and Zn-C60 distances are much longer than the Co-C60 one. The evaluated MP-C60 binding energies (Ebind) are 0.8 eV (18.5 kcal/mol) for M=Fe/Co and 0.5 eV (11.5 kcal/mol) for M=Ni/Cu/Zn (Ebind is about 0.2 eV larger in the case of C60-MTPP). They are believed to be reliable and accurate based on our dispersion-corrected DFT calculations that included the counterpoise (CP) correction. The effects of the C60 contact on the redox properties of MP were also examined. PMID- 17417892 TI - Hybridization detection of DNA by measuring organic small molecule amplified resonance light scattering signals. AB - The interaction of organic small molecules (OSMs) with a biological molecule is very important. In this contribution, quinone-imine dyes including Acridine Yellow (AY), Neutral Red (NR), Acridine Orange (AO), Brilliant Cresyl Blue (BCB), Thionin (TN), Azur A (AA), Azur B (AB), and Methylene Blue (MB) respectively with double strand DNA (dsDNA) and single strand DNA (ssDNA) were investigated based on the measurements of enhanced resonance light scattering (RLS) and TEM. Mechanism investigations have shown that groove binding occurs between dsDNA and these OSMs, which depends on G-C sequences of dsDNA and the volumes of OSMs. With the amplified RLS signals resulting from the interactions of OSMs with DNA, a new technique has been proposed to detect the hybridization and mismatch of DNA labeling neither the target nor the probe DNA. The results have suggested that the extent of the amplified RLS signals of dsDNA by AY is the maximum among these eight OSMs, and therefore, it has been selected as a typical model system for further discussions. PMID- 17417893 TI - Watching ultrafast barrierless excited-state isomerization of pseudocyanine in real time. AB - The photoinduced excited-state processes in 1,1'-diethyl-2,2'-cyanine iodine are investigated using femtosecond time-resolved pump-probe spectroscopy. Using a broad range of probe wavelengths, the relaxation of the initially prepared excited-state wavepacket can be followed down to the sink region. The data directly visualize the directed downhill motion along the torsional reaction coordinate and suggest a barrierless excited-state isomerization in the short chain cyanine dye. Additionally, ultrafast ground-state hole and excited-state hole replica broadening is observed. While the narrow excited-state wavepacket broadens during pump-probe overlap, the ground-state hole burning dynamics takes place on a significantly longer time-scale. The experiment reported can be considered as a direct monitoring of the shape and the position of the photoprepared wavepacket on the excited-state potential energy surface. PMID- 17417894 TI - Dynamics of solvent and rotational relaxation of coumarin-153 in room-temperature ionic liquid 1-butyl-3-methyl imidazolium tetrafluoroborate confined in poly(oxyethylene glycol) ethers containing micelles. AB - We have investigated solvent and rotational relaxation of coumarin 153 (C-153) in room-temperature ionic liquid (RTILs) 1-butyl-3-methyl-imidazolium tetrafluoroborate ([bmim][BF(4)]) and the ionic liquid confined in alkyl poly(oxyethylene glycol) ethers containing micelles. We have used octaethylene glycol monotetradecyl ether (C(14)E(8)) and octaethylene glycol monododecyl ether (C(12)E(8)) as surfactants. In the [bmim][BF(4)]-C(14)E(8) micelle, we have observed only a 22% increase in solvation time compared to neat [bmim][BF(4)], whereas in the [bmim][BF(4)]-C(12)E(8) system, we have observed approximately 57% increase in average solvation time due to micelle formation. However, the slowing down in solvation time on going from neat RTIL to RTIL-confined micelles is much smaller compared to that on going from water to water confined micellar aggregates. The 22-57% increase in solvation time is attributed to the slowing down of collective motions of cations and anions in micelles. The rotational relaxation times become faster in both the micelles compare to neat [bmim][BF(4)]. PMID- 17417895 TI - Intermolecular interactions and dynamics of room temperature ionic liquids that have silyl- and siloxy-substituted imidazolium cations. AB - The intermolecular interactions and dynamics of novel ionic liquids with alkylsilyl and alkylsiloxy substitutions on the cations are studied by measuring the intermolecular vibrational spectra and reorientational dynamics using femtosecond Kerr effect methods. The new ionic liquids include 1 dimethylphenylsilylmethyl-3-methylimidazolium (PhSi-mim+), and 1-methyl-3 pentamethyldisiloxymethylimidazolium (SiOSi-mim+) cations paired with the bis(trifluoromethylsulfonyl)imide (NTf(2)-) anion. Measured ionic liquid viscosities are surprisingly low for such bulky cation substituents. DFT electronic structure calculations on the isolated ions provide additional information about the electrostatic interactions. PMID- 17417896 TI - Protic ionic liquids: preparation, characterization, and proton free energy level representation. AB - We give a perspective on the relations between inorganic and organic cation ionic liquids (ILs), including members with melting points that overlap around the borderline 100 degrees C. We then present data on the synthesis and properties (melting, boiling, glass temperatures, etc.) of a large number of an intermediate group of liquids that cover the ground between equimolar molecular mixtures and ILs, depending on the energetics of transfer of a proton from one member of the pair to the other. These proton-transfer ILs have interesting properties, including the ability to serve as electrolytes in solvent-free fuel cell systems. We provide a basis for assessing their relation to aprotic ILs by means of a Gurney-type proton-transfer free energy level diagram, with approximate values of the energy levels based on free energy of formation and pK(a) data. The energy level scheme allows us to verify the relation between solvent-free acidic and basic electrolytes, and the familiar aqueous variety, and to identify neutral protic electrolytes that are unavailable in the case of aqueous systems. PMID- 17417897 TI - Activation of p300 histone acetyltransferase by small molecules altering enzyme structure: probed by surface-enhanced Raman spectroscopy. AB - Reversible acetylation of nucleosomal histones and nonhistone proteins play pivotal roles in the regulation of all the DNA templated phenomenon. Dysfunction of the enzymes involved in the acetylation/deacetylation leads to several diseases. Therefore, these enzymes are the targets for new generation therapeutics. Here, we report the synthesis of trifluoromethyl phenyl benzamides and their effect on histone acetyltransferase (HAT) activity of p300. One of these benzamides, CTPB (N-(4-chloro-3-trifluoromethyl-phenyl)-2-ethoxy-6 pentadecyl-benzamide), was discovered as a potent activator of the p300 HAT activity. We have found that pentadecyl hydrocarbon chain of CTPB is required to activate the HAT only under certain context. Furthermore, our results show that the relative position of -CF3 and -Cl in CTB (N-(4-chloro-3-trifluoromethyl phenyl)-2-ethoxy-benzamide) is also very critical for the activation. Surface enhanced Raman spectroscopy (SERS) of p300 and the HAT activator complexes evidently suggest that the activation of HAT activity is achieved by the alteration of p300 structure. Therefore, apart from elucidating the chemical basis for small molecule mediated activation of p300, this report also describes, for the first time, Raman spectroscopic analysis of the complexes of histone modifying enzymes and their modulators, which may be highly useful for therapeutic applications. PMID- 17417898 TI - Interpolymer complexes based on the core/shell micelles. interaction of polystyrene-block-poly(methacrylic acid) micelles with linear poly(2 vinylpyridine) in 1,4-dioxane water mixtures and in aqueous media. AB - The size and structural changes of nanoparticles formed after the addition of poly(2-vinylpyridine), PVP, to block copolymer micelles of polystyrene-block poly(methacrylic acid), PS-PMA, were studied by light scattering and atomic force microscopy. Due to the strong hydrogen bonding between PVP and PMA segments, complex structures based on the core/shell micelles form in mixed selective solvents. As proven by a combination of light scattering and atomic force microscopy, individual PS-PMA micelles are "glued" together by PVP chains. The dialysis against solvents with a high content of water results in transient increase in polydispersity and turbidity of originally clear solutions. However, the precipitated polymer material dissolves in basic buffers and stable soluble nanoparticles reform in aqueous media. The behavior of their solutions was studied in a broad pH range by light scattering, atomic force microscopy and capillary zone electrophoresis. PMID- 17417899 TI - Structures of linear poly(ethylene oxide) compounds and potassium complexes in dichloromethane. AB - The H-1 NMR spectra of a series of linear poly(ethylene oxides) compounds (POE compounds) were measured in dichloromethane-d2 at 25 degrees C, where the POE compounds (HO-(CH2CH2O-)n-R) were unsubstituted POE, HEOn (R=H, n=3, 4, 6), and alkyl-substituted POE, DEOn (R=C12H25, n=4, 6, 8) and MeEO6 (R=CH3, n=6). All the peaks of H-1 NMR signals were assigned to each methylene proton of POE. The chemical shifts and coupling constant between vicinal protons were evaluated by a complete spin analysis. The spectral changes of POE compounds by the addition of potassium ion were measured at various metal-to-POE ratios. The chemical shift change of each methylene proton by the formation of the complex was evaluated. The downfield shift of methylene protons caused by the complex formation indicates that the ethylene oxide that the ethylene oxide moiety is coordinating to surround the potassium ion in the same manner as the cyclic crown ether complexes. The results of spin lattice relaxation time measurements of DEO6 suggest that all the methylenes of the ethylene oxides are immobilized by the coordination to the metal ion. Thus, it was confirmed that all oxygens of POE are participating in the complex formation. PMID- 17417900 TI - Insights into the structure and dynamics of a room-temperature ionic liquid: ab initio molecular dynamics simulation studies of 1-n-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]) and the [bmim][PF6]-CO2 mixture. AB - Ab initio molecular dynamics (AIMD) studies have been carried out on liquid 1-n butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]) and its mixture with CO2 using the Car-Parrinello molecular dynamics (CPMD) method. Results from AIMD and empirical potential molecular dynamics (MD) have been compared and were found to differ in some respects. With a strong resemblance to the crystal, the AIMD simulated neat liquid exhibits many cation-anion hydrogen bonds, a feature that is almost absent in the MD results. The anions were observed to be strongly polarized in the condensed phase. The addition of CO2 increased the probability of this hydrogen bond formation. CO2 molecules in the vicinity of the ions of [bmim][PF6] exhibit larger deviations from linearity in their instantaneous configurations. The polar environment of the liquid induces a dipole moment in CO2, lifting the degeneracy of its bending mode. The calculated splitting in the vibrational mode compares well with infrared spectroscopic data. The solvation of CO2 in [bmim][PF6] is primarily facilitated by the anion, as seen from the radial and spatial distribution functions. CO2 molecules were found to be aligned tangential to the PF6 spheres with their most probable location being the octahedral voids of the anion. The structural data obtained from AIMD simulations can serve as a benchmark to refine interaction potentials for this important room temperature ionic liquid. PMID- 17417901 TI - Virial coefficients and demixing of athermal nonadditive mixtures. AB - We compute the fourth virial coefficient of a binary nonadditive, hard-sphere mixture over a wide range of deviations from diameter additivity and size ratios. Hinging on this knowledge, we build up a y expansion (Barboy, B.; Gelbart, W. N. J. Chem. Phys. 1979, 71, 3053) in order to trace the fluid-fluid coexistence lines, which we then compare with the available Gibbs-ensemble Monte Carlo data and with the estimates obtained through two refined integral-equation theories of the fluid state. We find that in a regime of moderately negative nonadditivity and largely asymmetric diameters, relevant to the modeling of sterically and electrostatically stabilized colloidal mixtures, the fluid-fluid critical point is unstable with respect to crystallization. PMID- 17417902 TI - H2O3 as a reactive oxygen species: formation of 8-oxoguanine from its reaction with guanine. AB - Reaction of guanine with H2O3 in the absence and presence of a water molecule leading to the formation of 8-oxoguanine (8-oxoG) was investigated. Initial calculations were performed using imidazole (Im) as a model for the five-membered ring of guanine. The reactant, intermediate, and product complexes as well as transition states were obtained in gas phase at the B3LYP/6-31+G* and B3LYP/AUG cc-pVDZ levels of theory. In all the cases, except for the reactions involving imidazole, single-point energy calculations were performed in gas phase at the MP2/AUG-cc-pVDZ level of theory. Solvation calculations in aqueous media were carried out using the polarizable continuum model (PCM) of the self-consistent reaction field (SCRF) theory. Vibrational frequency analysis and intrinsic reaction coordinate (IRC) calculations were performed to ensure that the transition states connected the reactant and product complexes properly. Zero point energy (ZPE)-corrected total energies and Gibbs free energies at 298.15 K in gas phase and aqueous media were obtained. When a reaction of H2O3 in place of H2O2 with guanine is considered, the major barrier energy which is encountered at the first step is almost halved showing that H2O3 would be much more reactive than H2O2. Considering the reaction schemes investigated here and the observed fact that H2O3 is dissociated easily under ambient conditions, it appears that H2O3 would serve as an effective reactive oxygen species. PMID- 17417903 TI - Design for hydrogen storage materials via observation of adsorption sites by computer tomography. AB - An effective method denoted as "computer tomography for materials" (mCT) was employed to study the adsorption sites inside metal-organic frameworks (MOFs) at any positions and any view angles. For MOF-5, the first adsorption site alpha( COO)3 was clearly observed from the mCT images; it locates at the position where three -COO groups joined like a cup. There are four alpha(-COO)3 sites around the Zn4O cluster. Two of them located at the diagonal of the Zn4O cluster are in the same plane "A", whereas the other two equivalent adsorption sites are in another plane "B", which is about 5.4 A away from the plane A. It was found that the electronegativity of oxygen atoms is very important to the adsorption of hydrogen molecules. The hydrogen amount adsorbed in MOFs might be enhanced by introducing some strong electronegative atoms to the organic linkers or frameworks. On the basis of this point of view, five new MOF materials were designed. The adsorbed amounts both in number of hydrogen molecules per unit cell and weight uptake for all of the designed MOFs were calculated. The adsorption amounts of designed MOFs were improved, and the amount for MOF-d5 at 1 bar is as high as 3.7 wt %. It is nearly 5-6 times of that of MOF-5 as a whole. It can be observed that extra adsorption sites were formed in the pores and the effective occupation rate of pore space was obviously improved viewing from the mCT images. These results may give helpful suggestions for the synthetic experimentalists. PMID- 17417904 TI - Tuning supramolecular structuring at the nanoscale level: nonstoichiometric soluble complexes in dilute mixed solutions of alginate and lactose-modified chitosan (chitlac). AB - Two oppositely charged polysaccharides, alginate and a lactose-modified chitosan (chitlac), have been used to prepare dilute binary polymer mixtures at physiological pH (7.4). Because of the negative charge on the former polysaccharide and the positive charge on the latter, polyanion-polycation complex formation occurred. A complete miscibility between the two polysaccharides was attained in the presence of both high (0.15 M) and low (0.015 M) concentrations of simple 1:1 supporting salt (NaCl), as confirmed by turbidity measurements; phase separation occurred for intermediate values of the ionic strength (I). The binary solutions were further characterized by means of light scattering, specific viscosity, and fluorescence quenching measurements. All of these techniques pointed out the fundamental role of the electrostatic interactions between the two oppositely charged polysaccharides in the formation of nonstoichiometric polyelectrolyte soluble complexes in dilute solution. Fluorescence depolarization (P) experiments showed that the alginate chain rotational mobility was impaired by the presence of the cationic polysaccharide when 0.015 M NaCl was used. Moreover, upon addition of calcium, the P values of the binary polymer mixture in 0.015 M NaCl increased more rapidly than that of an alginate solution without chitlac, suggesting an efficient crowding of the negatively charged alginate chains caused by the polycation. PMID- 17417905 TI - Cationic amphiphilic model networks based on symmetrical ABCBA pentablock terpolymers: synthesis, characterization, and modeling. AB - Eight isomeric networks based on equimolar terpolymers were synthesized using group transfer polymerization (GTP) and were characterized in terms of their swelling properties. Two hydrophilic monomers, the nonionic methoxy hexa(ethylene glycol) methacrylate (HEGMA) and the ionizable 2-(dimethylamino)ethyl methacrylate (DMAEMA), and a hydrophobic (nonionic) monomer, methyl methacrylate (MMA), were employed for the syntheses. 1,4 Bis(methoxytrimethylsiloxymethylene)cyclohexane (MTSMC) was used as the bifunctional GTP initiator, while ethylene glycol dimethacrylate (EGDMA) served as the cross-linker. Seven of the networks were model networks, six of which were based on the symmetrical pentablock terpolymers ABCBA, ACBCA, BACAB, BCACB, CBABC, and CABAC, whereas the seventh model network was based on the statistical terpolymer. The eighth network was a randomly cross-linked network based on the statistical terpolymer, prepared by the simultaneous quaterpolymerization of the three monomers and the cross-linker. The molecular weights and molecular weight distributions of the linear pentablock terpolymer precursors, as well as those of their homopolymer and ABA triblock copolymer precursors, were characterized by gel permeation chromatography (GPC) in tetrahydrofuran. The sol fraction of each network was measured and found to be relatively low. The aqueous degrees of swelling of all networks were found to increase at acidic pH due to the ionization of the DMAEMA tertiary amine units. The acidic degrees of swelling of the pentablock terpolymer networks were lower than those of their statistical counterparts due to microphase separation in the former type of networks, also confirmed by thermodynamic calculations and small-angle neutron scattering experiments. PMID- 17417906 TI - Antibacterial polypropylene via surface-initiated atom transfer radical polymerization. AB - Polypropylene (PP) coated by a non-leachable biocide was prepared by chemically attaching poly(quaternary ammonium) (PQA) to the surface of PP. The well-defined poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA), a precursor of PQA, was grown from the surface of PP via atom transfer radical polymerization (ATRP). The tertiary ammine groups in PDMAEMA were consequently converted to QA in the presence of ethyl bromide. Successful surface modification was confirmed by ATR FTIR, contact angle measurement, and an antibacterial activity test against Escherichia coli (E. coli). The biocidal activity of the resultant surfaces depends on the amount of the grafted polymers (the number of available quaternary ammonium units). With the same grafting density, the surface grafted with relatively high MW polymers (M(n) > 10,000 g/mol) showed almost 100% killing efficiency (killing all of the input E. coli (2.9 x 10(5)) in the shaking test), whereas a low biocidal activity (85%) was observed for the surface grafted with shorter PQA chains (M(n) = 1,500 g/mol). PMID- 17417907 TI - Cytotoxic withanolides from Tubocapsicum anomalum. AB - Fifteen new withanolides (1-8, 11-17) and two known withanolides, withanolide D (9) and 17alpha-hydroxywithanolide D (10), were isolated from the stems, roots, and leaves of Tubocapsicum anomalum using bioassay-directed fractionation. The structures were determined by spectroscopic and chemical methods, and the absolute configurations were established by CD analysis and by the Mosher ester method. The structure of 1 and 3 were further confirmed by X-ray crystallographic analysis. Compounds 1, 4-6, 8-10, and 13 showed significant cytotoxic activity against Hep G2, Hep 3B, A-549, MDA-MB-231, MCF-7, and MRC-5 cell lines. PMID- 17417908 TI - Ecdysteroids from Serratula wolffii roots. AB - Two new natural ecdysteroids, 20,22-didehydrotaxisterone (1) and 1-hydroxy-20,22 didehydrotaxisterone (2), were isolated from the roots of Serratula wolffii. Their structures were elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. The biological activities of these compounds were determined via oral aphid (Acyrthosiphon pisum (Harris)) tests. Compound 1 was inactive and compound 2 exhibited very low toxicity in the oral aphid test. The activities of these two ecdysteroids were in agreement with those of other 22 deoxyecdysteroids. PMID- 17417909 TI - Expeditious access to unprotected racemic pyroglutamic acids. AB - A series of biologically intriguing pyroglutamic acids were synthesized in racemic form by employing indole-isonitrile and ammonium acetate in the Ugi 4 center-3-component reaction of gamma-ketoacids. PMID- 17417910 TI - A practical Buchwald-Hartwig amination of 2-bromopyridines with volatile amines. AB - A practical Buchwald-Hartwig amination of 2-bromopyridines with volatile amines is developed in sealed tubes. The method provides an expedient entry to a variety of secondary and tertiary aminopyridines that are otherwise not readily synthesized. PMID- 17417911 TI - Multistep synthesis on SU-8: combining microfabrication and solid-phase chemistry on a single material. AB - SU-8 is an epoxy-novolac resin and a well-established negative photoresist for microfabrication and microengineering. The photopolymerized resist is an extremely highly crosslinked polymer showing outstanding chemical and physical robustness with residual surface epoxy groups amenable for chemical functionalization. In this paper we describe, for the first time, the preparation and surface modification of SU-8 particles shaped as microbars, the attachment of appropriate linkers, and the successful application of these particles to multistep solid-phase synthesis leading to oligonucleotides and peptides attached in an unambiguous manner to the support surface. PMID- 17417912 TI - Solid-phase synthesis of a thymidinyl dipeptide urea library. AB - A thymidinyl dipeptide urea library with structural similarity to the nucleoside peptide class of antibiotics was designed and synthesized. To generate the library, a solid-phase synthesis was developed starting from 5'-azidothymidine attached to a polystyrene butyl diethylsilane (PS-DES) resin support. This study describes the prelibrary solid-phase synthesis development including maximum loading capacity optimization, selection of orthogonal functionalized side-chain protection strategies, synthesis of a 64-member test library, and optimization of the final cleavage step. Using the optimized procedures, we synthesized a 1000 member library in a 50 micromol quantity using IRORI-directed sorting technology in MiniKans, producing the target library in good yields and purity. PMID- 17417913 TI - Electronic and structural characteristics of zinc-blende wurtzite biphasic homostructure GaN nanowires. AB - We report a new biphasic crystalline wurtzite/zinc-blende homostructure in gallium nitride nanowires. Cathodoluminescence was used to quantitatively measure the wurtzite and zinc-blende band gaps. High-resolution transmission electron microscopy was used to identify distinct wurtzite and zinc-blende crystalline phases within single nanowires through the use of selected area electron diffraction, electron dispersive spectroscopy, electron energy loss spectroscopy, and fast Fourier transform techniques. A mechanism for growth is identified. PMID- 17417914 TI - Quantitative proteome analysis of barley seeds using ruthenium(II)-tris (bathophenanthroline-disulphonate) staining. AB - This paper describes the application of the recently introduced fluorescence stain Ruthenium(II)-tris-(bathophenanthroline-disulphonate) (RuBP) on a comparative proteome analysis of two phenotypically different barley lines. We carried out an analysis of protein patterns from 2-D gels of the parental lines of the Oregon Wolfe Barley mapping population DOM and REC and stained with either the conventional colloidal Coomassie Brilliant Blue (cCBB) or with the novel RuBP solution. We wished to experimentally verify the usefulness of such a stain in evaluating the complex pattern of a seed proteome, in comparison to the previously used cCBB staining technique. To validate the efficiency of visualization by both stains, we first compared the overall number of detected protein spots. On average, 790 spots were visible by cCBB staining and 1200 spots by RuBP staining. Then, the intensity of a set of spots was assessed, and changes in relative abundance were determined using image analysis software. As expected, staining with RuBP performed better in quantitation in terms of sensitivity and dynamic range. Furthermore, spots from a cultivar-specific region in the protein map were chosen for identification to asses the gain of biological information due to the staining procedure. From this particular region, eight spots were visualized exclusively by RuBP and identification was successful for all spots, proving the ability to identify even very low abundant proteins. Performance in MS analysis was comparable for both protein stains. Proteins were identified by MALDI-TOF MS peptide mass fingerprinting. This approach was not successful for all spots, due to the restricted entry number for barley in the database. Therefore, we subsequently used LC-ESI-Q-TOF MS/MS and de novo sequencing for identification. Because only an insufficient number of proteins from barley is annotated, an EST-based identification strategy was chosen for our experiment. We wished to test whether under these limitations the application of a more sensitive stain would lead to a more advanced proteome approach. In summary, we demonstrate here that the application of RuBP as an economical but reliable and sensitive fluorescence stain is highly suitable for quantitative proteome analysis of plant seeds. PMID- 17417915 TI - Supramolecular photochirogenesis with biomolecules. Mechanistic studies on the enantiodifferentiation for the photocyclodimerization of 2-anthracenecarboxylate mediated by bovine serum albumin. AB - Photophysics and photochemistry of 2-anthracenecarboxylate (AC) bound to bovine serum albumin (BSA) were investigated in detail for the first time by electronic absorption, circular dichroism (CD), steady-state and time-resolved fluorescence, fluorescence quenching, and product analysis studies. Through the spectroscopic investigations, it was revealed that the four independent binding pockets of BSA, which are known to accommodate 1, 3, 2, and 3 AC molecules in the order of decreasing affinity, are distinctly different in hydrophobicity, chiral environment, and accessibility. Interestingly, AC bound to site 1 gave highly structured fluorescence with dual lifetimes of 4.8 and 2.1 ns in an intensity ratio of 3:2, which may be assigned to the existence of two positional or orientational isomers within the very hydrophobic site 1. In contrast, the lifetime of AC in site 2 was much longer (13.3 ns), and ACs in sites 3 and 4 have broader fluorescence spectra with lifetimes that were practically indistinguishable from that in bulk water (15.8 ns). Although each of sites 2-4 simultaneously binds multiple AC molecules, no CD exciton coupling or static fluorescence quenching was detected, indicating that ACs bound to each site are not in close proximity to each other. Quenching studies with nitromethane further confirmed the significant difference in accessibility among the binding sites; thus, ACs bound to sites 1 and 2 are highly protected from the attack of the quencher, affording 32 and 10 times smaller rate constants than that for free AC in water. Product studies in the presence and absence of nitromethane more clearly revealed the photochirogenic performance of each binding site. Although the addition of nitromethane did not greatly alter the product distribution, the enantiomeric excesses (ee's) of chiral cycloadducts 2 and 3 were critically manipulated by selectively retarding the photoreaction occurring at the more accessible binding sites. Thus, the highest ee of 38% was obtained for 2 in the presence of 18 mM nitromethane, while the highest ee of 58% was attained for 3 in the absence of nitromethane, both at [AC]/[BSA]=3.6. PMID- 17417916 TI - [Helicobacter pylori infection: antibiotic resistance]. PMID- 17417917 TI - [Helicobacter pylori erradication and its relation to antibiotic resistance and CYP2C19 status]. AB - OBJECTIVE: to assess the efficacy of rabeprazole (RPZ), amoxicillin (Am), and clarithromycin (Cla) (7 vs. 14 days) in the eradication of H. pylori, and to determine the effect of strain-specific antibiotic resistance and host CYP2C19 status. MATERIAL AND METHODS: first, we determined the CYP2C19 status of 100 healthy subjects to establish a sample size for the clinical trial. Then, 59 H. pylori-infected patients were randomized to receive RPZ (20 mg daily) plus Cla (500 mg b.d.) and Am (1,000 mg b.d.) for 7 vs. 14 days. The MIC for Am and Cla were determined using the agar dilution method. The CYP2C19 genotype was determined by the PCR-RFLP method. RESULTS: In the per-protocol analysis (PP) eradication rates were 89.7 and 72% for the 7- and 14-day groups (p = 0.159). In the intention to-treat analysis (ITT) eradication rates were 86.7 and 62.1% in the 7- and 14-day groups, respectively (p = 0.06). None of the strains was resistant to Am, and 4 strains were resistant to Cla: 3 (11.1%) in the 14-day group and 1 (4%) in the 7-day group. Neither strain-specific antibiotic resistance nor host CYP2C19 status influenced eradication rates. CONCLUSIONS: both 7- and 14-day therapies were effective for H. pylori eradication. Strain resistance and CYP2C19 status do not seem to influence eradication rates in the studied population. PMID- 17417918 TI - [Role of malnutrition in intestinal anastomosis collagenization: an analysis of procollagen (PINP) and carboxyterminal telopeptide (ICTP) by radioimmunoassay]. AB - INTRODUCTION: some clinical, anatomo-pathological, and technical factors influence the correct healing of intestinal suture following an intestinal resection. One of the most influential factors is patient nutritional status. OBJECTIVES: to evaluate the influence of malnutrition on the viability of primary intestinal anastomosis by the analysis of collagen I deposition. METHODS: 40 Wistar rats, radioimmunoassay material. We used 2 groups of rats, 20 animals in each group: a control group (A) and a "malnutrition" group (B). RESULTS: there was a decrease in PINP (procollagen) deposition in the colon of group B rats as compared to the colon of group A (0.3620 and 0.4340 mg/g respectively) (p = 0.032). There is an increase in ICTP (carboxyterminal telopeptide) in the colon of group B (0.9545 as against 0.8460 mg/g in group A) (p = 0.875). In anastomoses of group B there was a decrease in PINP synthesis as compared to group A (0.376 and 0.468 mg/g respectively, p = 0.002). As regards ICTP, there was an increase in group B (p = 0.330). In relation to the control group no differences were observed in ICTP increases in group B (p = 1). CONCLUSIONS: colonic anastomosis increases the levels of PINP and ICTP in healed tissue (p = 0.000); malnutrition reduces collagenization in anastomoses (p = 0.000). PMID- 17417919 TI - [Perforation risk in esophageal endoscopic mucosal resection with ligation: an experimental study with two ligator models]. AB - OBJECTIVE: endoscopic mucosal resection with ligation (EMRL) is considered an efficient, safe method for the treatment of some esophageal, gastric and colorectal tumors. We conducted this study using a porcine model in order to compare the safety of esophageal EMRL with two multiband ligation systems, since many centers only use these ligator models in EMRL (commercialized for varix ligation). METHODS: eight pigs were used, which were submitted to 23 esophageal resections without previous injection. Ten resections were conducted using the Six Shooter Saeed model, and 13 resections used the Speedband Superview Super 7 model. The technique was also compared by making random cuts either above or below the band. RESULTS: five perforations occurred, all of them using the Speedband model. This represents 38.5% of total in the Speedband model group. No perforation occurred when using the Six Shooter model (p = 0.046). On the contrary, no significant differences were found regarding frequency of perforation when cutting above or below the band. CONCLUSIONS: esophageal EMRL using the Speedband model without previous injection leads to perforation in a high percentage of cases in an experimental animal model. Further studies are required to find out whether a previous injection may increase the safety of this technique with this ligator model. PMID- 17417920 TI - [Serologic diagnosis of Helicobacter pylori in endoscopy personnel. Serology in endoscopists]. AB - OBJECTIVE: the Helicobacter pylori entered into the medical world, particularly in the field of the Gastroenterology, revolutionizing it, and changing partially our physiopathological concepts and treatment of the gastroduodenal ulcer. This infestation results in an immune response, that is measured by means of serologic tests, which are very useful in the massive screening studies, mainly in population at risk like those who are institutionalized and the personnel of health, particularly workers of the departments of Gastroenterology. METHOD: motivated for this, we carry out a prospective study in 38 endoscopists using as controls 38 workers not related with a Gastroenterology department. Thus, we analyzed the presence of antibodies against Helicobacter pylori among the personnel of the departments of Gastroenterology of three hospitals of Havana City. These personnel filled a form designed for this purpose containing following data: age, sex, time of work as endoscopist, use of protection means, and history of gastroduodenal pathology. RESULTS: there were 24 women in each group (63,2%). Whereas the seroprevalence among endoscopists was 39,47%, in the control group, only three people were seropositives (7,89%). We found a positive correlation between the time working as endoscopists and the rate of seropositivity. CONCLUSIONS: our study demonstrated that endoscopists have a significantly higher risk than the controls of contracting the infection by the Helicobacter pylori, and that the seropositive endoscopists have sintomatology more frequently than the seronegative endoscopists, although these differences were not statistically significant. PMID- 17417921 TI - Energy change in the formation of conjugated bilirubin: a possible responsive mechanism for liver cell pathology. AB - Bilirubin is formed when red blood cells die and their hemoglobin is broken down within the macrophages into heme and globins. In the liver, bilirubin is conjugated with UDP-glucuronate, making it water-soluble diglucuronide. Concerning this conjugation, a molecule of bilirubin reacts with two molecules of glucoronic acid. However, the nature of this energy-consuming reaction in the formation of conjugated bilirubin has never been reported, and this can be important for its potential implication in hyperbilirubinemia. In this work, the author calculated the energy required by conjugated-bilirubin formation per molecule. The energy required for complex formation is -22 kCal/mol. The nature of this energy-producing reaction can be a good explanation. Increased energy delivery in conjugated hyperbilirubinemia in liver disease might be a responsive mechanism to hepatic damage. PMID- 17417922 TI - Pneumatosis intestinalis and hepatic-portal-mesenteric venous gas in intestinal ischemia. AB - OBJECTIVE: the purpose of this study was to report the authors experience in the diagnosis, management, and clinical outcome of intestinal ischemia with pneumatosis intestinalis (PI) and hepatic-portal-mesenteric venous gas (HPMVG). EXPERIMENTAL DESIGN: a retrospective analysis. METHODS: the medical records of 12 cases of intestinal ischemia with PI and HPMVG spanning the last 10 years were reviewed. Clinical characteristics, imaging study results, surgical findings, histological findings, management, and clinical outcome were evaluated. RESULTS: of the 12 cases that were evaluated, 9 patients presented with abdominal distension, three, with abdominal pain, five, with shock on admission, and six, with fever. Most patients (83%) had a history of diabetes mellitus. Dilated loops of bowel were identified on plain film radiography in 9 of 12 of patients. Only 3 cases were initially suspected of having abnormal gas over the RUQ or intramural intestine. All patients underwent abdominal computed tomography (CT). Nine patients were diagnosed with ischemic changes of the intestine (2 cases had colon involvement), and 3 patients were found to have a gangrenous intestine. While all patients underwent segmental resection of the intestine, ten of 12 patients died post-operatively. CONCLUSIONS: abdominal CT findings of PI and HPMVG in intestinal ischemia do not appear to either directly correlate with the gross findings or predict clinical outcome. The clinical outcome of patients affected with this condition was poor, even following surgical intervention. A positive clinical outcome may depend on the severity and extent of the underlying disease(s). PMID- 17417923 TI - [Chronic intestinal pseudo-obstruction]. AB - Chronic intestinal pseudo-obstruction (CIPO) is a syndrome characterized by the presence of recurrent episodes of clinical intestinal obstruction in the absence of obstructive lesions. Although this syndrome is rare, it causes a high morbidity. It is caused by a disturbance of the intestinal motility, that results in a failure of the progression of the intestinal content. Basically, the failure of the intestinal motility is a consequence of muscular disorder, neurological disorder or both. Usually, CIPO is secondary to other systemic disease; however, in the last years, many cases of primary CIPO have been described. The use of new manometric tecniques and specific histological procedures have allowed to clarify the pathogenesis of some of these entities including mitochondrial diseases and paraneoplasic syndromes. Clinical manifestations of CIPO are diverse, depending on the location and extension of the motility disorder. As the diagnosis of this disease is usually not an easy task, patients frecuently undergo unnecesary surgical interventions, are diagnosed of psyquiatric disorders, or the correct diagnosis is delayed several years after the first symptoms arise. The aims of the treatment are to maintain the nutritional condition and to improve symptoms using nutritional measures, drugs or, eventually, endoscopical or surgical procedures. PMID- 17417924 TI - [Massive mesenteric angiomatosis and low digestive hemorrhage in a patient with Klippel-Trenaunay-Weber syndrome]. PMID- 17417925 TI - [Benign intracranial hypertension and ulcerative colitis]. PMID- 17417926 TI - [Hepatoportal sclerosis, an unfrequent cause of portal hypertension]. PMID- 17417927 TI - [Cutaneous metastasis in a patient with colangiocarcinoma]. PMID- 17417928 TI - Synchronous small-cell lung cancer following colorectal carcinoma. An uncommon entity in the elderly. PMID- 17417929 TI - [Acute abdomen due to massive abdominal wall hematoma]. PMID- 17417930 TI - [Giant pneumoperitoneum secondary to percutaneous endoscopic gastrostomy]. PMID- 17417931 TI - State of diabetes care in the United States. AB - As of 2005, it was estimated that 7% of the US population, approximately 21 million people, have diabetes. The major concern with diabetes is long-term complications, which are responsible for increased rates of morbidity and mortality. Studies have shown that lower glycosylated hemoglobin reduces the microvascular and macrovascular complications associated with diabetes. To achieve this goal, the American Diabetes Association provides treatment goals to aggressively control diabetes to improve outcomes and decrease morbidity and mortality. Although studies have proved the beneficial effects of currently used agents, there are still various concerns, including weight gain, high risk of hypoglycemia, poor postprandial control, and failure to maintain long-term glycemic control. With the advent of new incretin-related therapies, some of these concerns may be addressed. Diabetes is of growing concern, and better knowledge of treatment options and goals should be a priority for all healthcare professionals. PMID- 17417932 TI - Guidelines and performance measures for diabetes. AB - Diabetes mellitus has become a major public health problem in the United States and around the world. A result of genetic predisposition combined with detrimental changes in lifestyle, type 2 diabetes is reaching epidemic proportions. The costs are very high, both in terms of financial burden and quality of life. A wide variety of healthcare organizations have developed diabetes guidelines to improve the quality of care for patients. Increasingly, there is agreement and collaboration on such guidelines, leading to the next step in quality management, performance measures. There is a growing trend to find ways to provide incentives for improved diabetes care. Initiatives such as the National Committee for Quality Assurance's physician recognition program, the Bridges to Excellence Diabetes Care Link program, the National Diabetes Quality Improvement Alliance, and the American Medical Association Physician Consortium for Performance Improvement are all helping to establish an incentive for improved diabetes care. PMID- 17417933 TI - New technologies and therapies in the management of diabetes. AB - Although there are numerous effective pharmacotherapeutic agents available to treat type 2 diabetes, 5% to 10% of the population with diabetes experience secondary failure. To help combat this issue, it is imperative that clinicians understand the limitations of some current therapies. Secondary failure can be due to decreasing beta cell function, poor adherence to treatment, weight gain, reduction of exercise, changes in diet, or illness. Glycemic control and cardiovascular risk reduction are of paramount concern; however, the nonglycemic effects of several new therapies to treat diabetes may be advantageous and positively affect the long-term cost of therapy. The discoveries of amylin and glucagonlike peptide-1 have furthered our understanding of the abnormalities involved in diabetes, enabling the development of additional therapeutic options. Incretin-based therapy, including incretin mimetics such as exenatide and the yet to-be-approved dipeptidyl peptidase-4 inhibitors, and new basal and inhaled insulin may change the way we currently treat type 2 diabetes. PMID- 17417934 TI - Economic and clinical impact of innovative pharmacy benefit designs in the management of diabetes pharmacotherapy. AB - Diabetes has enormous personal, economic, and societal costs. Despite the proven financial and clinical benefits of tight glycemic control, there remain a number of barriers to improving outcomes, including poor adherence to pharmacotherapy, which is responsible for inadequate control of diabetes and the associated financial burden. There are many factors that may affect an individual's adherence to a medication regimen, such as personal beliefs, the complexity of the regimen, and, most importantly, cost. Many benefit plans are affected by the rising cost incurred by its members, which could be potentially resolved if benefit plans think of innovative ways to promote adherence. One such example can be found in the benefit design chosen by Pitney Bowes. Their pharmacy benefit design contained diabetes medications in tier 1, which were under tier 2 in the past. Findings showed that as patient adherence increases with time, associated healthcare costs and annual pharmacy costs decrease. PMID- 17417935 TI - Fundamental components of attention. AB - A mechanistic understanding of attention is necessary for the elucidation of the neurobiological basis of conscious experience. This chapter presents a framework for thinking about attention that facilitates the analysis of this cognitive process in terms of underlying neural mechanisms. Four processes are fundamental to attention: working memory, top-down sensitivity control, competitive selection, and automatic bottom-up filtering for salient stimuli. Each process makes a distinct and essential contribution to attention. Voluntary control of attention involves the first three processes (working memory, top-down sensitivity control, and competitive selection) operating in a recurrent loop. Recent results from neurobiological research on attention are discussed within this framework. PMID- 17417936 TI - Orbitofrontal cortex and its contribution to decision-making. AB - Damage to orbitofrontal cortex (OFC) produces an unusual pattern of deficits. Patients have intact cognitive abilities but are impaired in making everyday decisions. Here we review anatomical, neuropsychological, and neurophysiological evidence to determine the neuronal mechanisms that might underlie these impairments. We suggest that OFC plays a key role in processing reward: It integrates multiple sources of information regarding the reward outcome to derive a value signal. In effect, OFC calculates how rewarding a reward is. This value signal can then be held in working memory where it can be used by lateral prefrontal cortex to plan and organize behavior toward obtaining the outcome, and by medial prefrontal cortex to evaluate the overall action in terms of its success and the effort that was required. Thus, acting together, these prefrontal areas can ensure that our behavior is most efficiently directed towards satisfying our needs. PMID- 17417937 TI - Trinucleotide repeat disorders. AB - The discovery that expansion of unstable repeats can cause a variety of neurological disorders has changed the landscape of disease-oriented research for several forms of mental retardation, Huntington disease, inherited ataxias, and muscular dystrophy. The dynamic nature of these mutations provided an explanation for the variable phenotype expressivity within a family. Beyond diagnosis and genetic counseling, the benefits from studying these disorders have been noted in both neurobiology and cell biology. Examples include insight about the role of translational control in synaptic plasticity, the role of RNA processing in the integrity of muscle and neuronal function, the importance of Fe-S-containing enzymes for cellular energy, and the dramatic effects of altering protein conformations on neuronal function and survival. It is exciting that within a span of 15 years, pathogenesis studies of this class of disorders are beginning to reveal pathways that are potential therapeutic targets. PMID- 17417938 TI - Visual perception and memory: a new view of medial temporal lobe function in primates and rodents. AB - The prevailing view of medial temporal lobe (MTL) function has two principal elements: first, that the MTL subserves memory but not perception, and second, that the many anatomically distinctive parts of the MTL function together in the service of declarative memory. Recent neuropsychological studies have, however, challenged both opinions. First, studies in rodents, nonhuman primates, and humans suggest that the perirhinal cortex represents information about objects for both mnemonic and perceptual purposes. Second, the idea that MTL components contribute to declarative memory in similar ways has also been contradicted. Whereas the perirhinal cortex plays an essential role in familiarity-based object recognition, the hippocampus contributes little, if at all, to this function. In both primates and rodents, the hippocampus contributes to the memory and perception of places and paths, whereas the perirhinal cortex does so for objects and the contents of scenes. PMID- 17417939 TI - The medial temporal lobe and recognition memory. AB - The ability to recognize a previously experienced stimulus is supported by two processes: recollection of the stimulus in the context of other information associated with the experience, and a sense of familiarity with the features of the stimulus. Although familiarity and recollection are functionally distinct, there is considerable debate about how these kinds of memory are supported by regions in the medial temporal lobes (MTL). Here, we review evidence for the distinction between recollection and familiarity and then consider the evidence regarding the neural mechanisms of these processes. Evidence from neuropsychological, neuroimaging, and neurophysiological studies of humans, monkeys, and rats indicates that different subregions of the MTL make distinct contributions to recollection and familiarity. The data suggest that the hippocampus is critical for recollection but not familiarity. The parahippocampal cortex also contributes to recollection, possibly via the representation and retrieval of contextual (especially spatial) information, whereas perirhinal cortex contributes to and is necessary for familiarity-based recognition. The findings are consistent with an anatomically guided hypothesis about the functional organization of the MTL and suggest mechanisms by which the anatomical components of the MTL interact to support the phenomenology of recollection and familiarity. PMID- 17417941 TI - Virus-induced disease: altering host physiology one interaction at a time. AB - Virus infections are the cause of numerous plant disease syndromes that are generally characterized by the induction of disease symptoms such as developmental abnormalities, chlorosis, and necrosis. How viruses induce these disease symptoms represents a long-standing question in plant pathology. Recent studies indicate that symptoms are derived from specific interactions between virus and host components. Many of these interactions have been found to contribute to the successful completion of the virus life-cycle, although the role of other interactions in the infection process is not yet known. However, all share the potential to disrupt host physiology. From this information we are beginning to decipher the progression of events that lead from specific virus host interactions to the establishment of disease symptoms. This review highlights our progress in understanding the mechanisms through which virus-host interactions affect host physiology. The emerging picture is one of complexity involving the individual effects of multiple virus-host interactions. PMID- 17417942 TI - Dual effect of butyrate on IL-1beta--mediated intestinal epithelial cell inflammatory response. AB - Butyrate (NaBu), a product of intestinal microbial metabolism, has been proposed as an anti-inflammatory agent for treating inflammatory bowel diseases. However, the molecular mechanisms implicated in the modulation of intestinal epithelial cell inflammatory response to NaBu remain unknown. Here, microarray analysis performed on nontransformed human crypt intestinal epithelial cells (HIEC) shows that NaBu regulated specifically the short-term IL-1beta -dependent induction of different inflammatory genes. While NaBu significantly increased the IL-1beta induction of genes like SAA2, C3, and IL-1alpha , other inflammatory genes like CXCL5, CXCL11, and IL-1beta were decreased. Induction of various genes such as CXCL8, CCL20, and IL-6 was unaffected by NaBu. We show that, compared to genes that are upregulated or downregulated by NaBu, genes that are unaffected by NaBu were induced more rapidly after IL-1beta treatment and contained a higher concentration of transcription factor binding sites in their promoter region. In addition, transient treatment with IL-1beta was sufficient for subsequent induction of NaBu-upregulated and NaBu-unaffected classes of genes, while a continuous presence of IL-1beta was required for NaBu-downregulated gene expression. In conclusion, our results suggest that fundamental differences predispose inflammatory genes to specific regulation by NaBu in intestinal epithelial cells, thereby allowing precise control of inflammation. PMID- 17417940 TI - Dynamic aspects of CNS synapse formation. AB - The mammalian central nervous system (CNS) requires the proper formation of exquisitely precise circuits to function correctly. These neuronal circuits are assembled during development by the formation of synaptic connections between thousands of differentiating neurons. Proper synapse formation during childhood provides the substrate for cognition, whereas improper formation or function of these synapses leads to neurodevelopmental disorders, including mental retardation and autism. Recent work has begun to identify some of the early cellular events in synapse formation as well as the molecular signals that initiate this process. However, despite the wealth of information published on this topic in the past few years, some of the most fundamental questions about how, whether, and where glutamatergic synapses form in the mammalian CNS remain unanswered. This review focuses on the dynamic aspects of the early cellular and molecular events in the initial assembly of glutamatergic synapses in the mammalian CNS. PMID- 17417943 TI - Plasmid DNA and IL-4 modulate expression of mHC class I and costimulatory molecules in B lymphocytes. AB - B lymphocytes are capable of spontaneous internalization of plasmid (p)DNA, an event that set in motion the antigen-presenting function in this class of hemopoietic cells. Previously, we showed that priming of CD8 T lymphocytes by spontaneously transgenic B lymphocytes requires T-cell help, and that this can be replaced by soluble IL-4. To better understand this phenomenon we studied the relative role of pDNA and IL-4 on the expression of MHC-I and a panel of critical costimulatory molecules (CD40, CD80, CD86, OX40L, and LAG-3). Whereas upregulation of MHC-I is contributed by pDNA, IL-4 mainly upregulates CD86 and to a lesser degree CD40. The two effects appear to be independent. In addition, however, it was found that IL-4 stabilizes MHC-I transcription in lymphocytes after spontaneous transgenesis with pDNA. These results further our understanding of events that take place in specialized mammalian cells after exposure to pDNA. They also point to the fact after pDNA internalization that the antigen presenting function of B lymphocytes can be complemented by IL-4, a cytokine normally produced by activated CD4 T cells. PMID- 17417944 TI - Nanosecond pulsed electric fields have differential effects on cells in the S phase. AB - Nanosecond pulsed electric fields (nsPEFs) are a type of nonthermal, nonionizing radiation that exhibit intense electric fields with high power, but low energy. NsPEFs extend conventional electroporation (EP) to affect intracellular structures and functions and depending on the intensity, can induce lethal and nonlethal cell signaling. In this study, HCT116 human colon carcinoma cells were synchronized to the S-phase or remained unsynchronized, exposed to electric fields of 60 kV/cm with either 60-ns or 300-ns durations, and analyzed for apoptosis and proliferative markers. Several nsPEF structural and functional targets were identified. Unlike unsynchronized cells, S-phase cells under limiting conditions exhibited greater membrane integrity and caspase activation and maintained cytoskeletal structure. Regardless of synchronization, cells exposed to nsPEFs under these conditions primarily survived, but exhibited some turnover and delayed proliferation in cell populations, as well as reversible increases in phosphatidylserine externalization, membrane integrity, and nuclei size. These results show that nsPEFs can act as a nonligand agonist to modulate plasma membrane (PM) and intracellular structures and functions, as well as differentially affect cells in the S-phase, but without effect on cell survival. Furthermore, nsPEF effects on the nucleus and cytoskeleton may provide synergistic therapeutic actions with other agents, such as ionizing radiation or chemotherapeutics that affect these same structures. PMID- 17417945 TI - Is low-frequency distribution of TGF-beta genotype associated with increased risk for end-stage renal disease? AB - End-stage renal disease has been associated with an inflammatory state. TGF-beta plays a critical role in antiinflammation counteracting inflammatory cytokines, wound healing, and tissue repair. We, therefore, speculated the protective role of TGF-beta in renal inflammation rather than inducing fibrosis. Three polymorphisms of TGF-beta (713-8delC), i.e., C deletion in intron sequence 8 base prior to exon-5 by PCR-RFLP and codon-10, Leu/Pro, and codon-25, Arg/Pro by Amplification Refractory Mutation System (ARMS-PCR) techniques were genotyped in 228 end-stage renal disease (ESRD) patients and 180 controls. Linkage disequilibrium (LD) and haplotype analysis was performed by Arlequin software. Our data showed positive association between codon-10 polymorphism and ESRD risk (P < 0.001; OR 4.845, 95% CI 2.57-9.11 for Pro/Pro). However, genotype frequencies were comparable in patients and controls for 713-8delC, while in the case of codon-25, a trend of higher frequency of Pro/Pro genotype (16.2% versus 10.0%) was observed but the P-value did not reach significant (P = 0.187). Significant association of codon-10 Pro/Pro was observed in patients with glomerulonephritis (P = 0.001; OR 4.138, 95%CI 2.1-8.13). LD was found significant between codon-10 and 25 (P = 0.021). Haplotype "Pro-Pro" showed 1.8 fold higher risk for ESRD (p = 0.003; OR = 1.867, 95%CI = 1.229-2.838). A combined analysis of the effect of TGF-beta (codon-10) with C-deletion and codon 25 showed significant difference for TGF-beta(10)-TGF-beta(C-del) (P = 0.010). In conclusion, the present study suggests that low-producing genotype (Pro/Pro) of TGF-beta (codon-10) polymorphism is associated with ESRD. Haplotype analysis further suggested that "Pro-Pro" (low producer) is associated with higher risk for ESRD. Thus, high-producing genotype of TGF-beta may be beneficial and may play a potential role in the resolution of renal inflammation. PMID- 17417946 TI - Gefitinib-sensitive EGFR lacking residues 746-750 exhibits hypophosphorylation at tyrosine residue 1045, hypoubiquitination, and impaired endocytosis. AB - Gefitinib-sensitive nonsmall cell lung cancers (NSCLC) are characterized by somatic mutations in the kinase domain of epidermal growth factor receptor (EGFR). The mutant EGFR forms are reported to mediate characteristic signal transduction pathways that are different from those mediated by the wild-type EGFR and are involved in transformation in vivo. We have examined signal transduction pathways initiated from a frequently identified gefitinib sensitizing mutant EGFR lacking residues 746-750 by employing a mouse fibroblast cell line that is free of endogenous EGFR and transiently transfected COS-7 cells. Upon EGF stimulation, the deletion-mutant EGFR mediated prolonged downstream signals. The analysis of the phosphotyrosine patterns of the receptor revealed that the deletion-mutant EGFR lacked phosphorylation at tyrosine residue 1045, which is the major binding site of Cbl. The EGF-induced endocytosis of the deletion-mutant EGFR was impaired. The ubiquitination and downregulation of the deletion-mutant EGFR were also reduced. On the other hand, another mutant, EGFR, possessing a L858R substitution, exhibited phosphorylation at 1045 and its downstream signalings were not prolonged. These data suggest that the signal transduction pathways initiated from these mutant forms are different, and that impaired endocytosis might be responsible for the prolonged signals mediated by the deletion-mutant EGFR. PMID- 17417947 TI - Contrasting impact of DNA repair gene XRCC1 polymorphisms Arg399Gln and Arg194Trp on the risk of lung cancer in the north-Indian population. AB - DNA repair forms the most effective defense system against DNA damage. The XRCC1 gene product is implicated in single-strand and base-excision repair mechanisms. Our main aim was to investigate the relationship between the XRCC1 gene with lung cancer on the north-Indian population. Blood samples from 225 North-Indian subjects including 103 newly diagnosed cases and 122 population-based healthy persons were collected. XRCC1 genotypes were detected using a PCR-RFLP technique. The data were analyzed by logistic regression analysis. XRCC1 polymorphisms at codon 399 were found to be protective in the development of lung cancer (OR--0.6, 95% CI--0.46-0.80, p-0.0008). The codon 194 Trp/Trp genotype was associated with a slightly increased risk of lung cancer. When assessed in nonsmokers, only the Arg/Trp genotype of XRCC1 codon 194 was positively associated with lung cancer (OR--2.3, 95% CI--0.77-7.20). Smoking also seemed to significantly interact with the combined genotypes of XRCC1 codon 399 Arg/Gln/Gln/Gln. In conclusion, the results have suggested that the XRCC1 gene might be the risk genotype for lung cancer in this population. PMID- 17417949 TI - Chondrogenic differentiation potential of human mesenchymal stem cells photoencapsulated within poly(ethylene glycol)-arginine-glycine-aspartic acid serine thiol-methacrylate mixed-mode networks. AB - Chondrogenesis of human mesenchymal stem cells (hMSCs) encapsulated in poly(ethylene glycol) (PEG)-based hydrogels was studied in the presence and absence of 5 ng/mL transforming growth factor beta and chondrogenic medium to better understand the role of the gel environment on this process. The lack of any cell-polymer interactions led to decreasing cell viability, as measured using adenosine triphosphate, over a 14-day period. The extent of chondrogenic differentiation was evaluated by immunostaining, and although viability dramatically decreased, cells cultured in chondrogenic differentiation medium expressed higher levels of collagen type II. Cells cultured in hMSC control medium remained undifferentiated and continued to express CD105, a MSC marker. To increase cell survival, arginine-glycine-aspartic acid-serine (RGDS) was incorporated into gels using a novel mixed-mode thiol-ene reaction by synthesizing a cysteine-cysteine-arginine-glycine-aspartic acid-serine-cysteine cysteine-glycine, N-terminus to C-terminus peptide sequence with pendant cysteine residues. A concentration of 5 mM RGDS incorporated into the network maintained 75% viability in control cultures. Further studies demonstrated that 5-mM RGDS chondrogenic cultures had greater gene expression for aggrecan and collagen II in conjunction with producing twice as much glycosaminoglycan as 0-mM chondrogenic cultures and 7 times that of control cultures. Incorporation of this peptide sequence not only allows for sustained viability, but also contributes to initiating chondrogenesis. PMID- 17417953 TI - Analysis of climatic predictions for extrinsic incubation of Dirofilaria in the United kingdom. AB - Transmission of dirofilariasis in Europe is dependent upon the presence of sufficient numbers of infected dogs, susceptible mosquitoes, and a suitable climate to permit extrinsic incubation of Dirofilaria in the mosquito intermediate host. Dogs returning to the United Kingdom from overseas have been infected with several vector-borne infections (Babesia, Ehrlichia, Leishmania, and Dirofilaria), and this paper assesses the climatic constraints on the potential rate of extrinsic incubation of Dirofilaria in the United Kingdom. A model using an established algorithm based on accumulated temperature predicts that summer temperatures during 1995-2000 may have been sufficient to permit complete incubation of Dirofilaria in 2 of the years for large parts of southern/central England, and 5 years around London. The occurrence of autochthonous transmission would be dependent upon additional factors related to frequency of returning infected dogs and the distribution and abundance of putative mosquito vectors, and these should be studied further. PMID- 17417954 TI - Bird species potentially involved in introduction, amplification, and spread of West Nile virus in a Mediterranean wetland, the Camargue (Southern France). AB - West Nile virus (WNV) is a mosquito-transmitted Flavivirus with a transmission cycle involving birds as amplifying hosts. Wild birds are also believed to carry WNV over large distances and are able to introduce it into new areas during migration and dispersal. In this paper, our objective is to provide lists of birds potentially involved in the introduction, the amplification and the spread of WNV in the Camargue, a Mediterranean wetland in the south of France where several WNV outbreaks have occurred since the 1960s. Bird species were classified according to the following ecological factors: migratory status and provenance area, used biotopes, abundance and period of presence in the Camargue. The obtained lists of bird species potentially involved in the introduction, amplification and spread of WNV should prove useful to determine target species on which further studies on WNV ecology in birds could be focused. PMID- 17417955 TI - Effect of gender on clinical and epidemiologic features of Lyme borreliosis. AB - The aim is to highlight the influence of patients' gender on Lyme borreliosis and especially erythema migrans (EM), focusing on exposure to tick bites, epidemiology, and the clinical picture. All studies were conducted in the county of Blekinge, located in southeastern Sweden. A prospective study was conducted in 235 individuals (women, n=110; men, n=125) engaged in recreational or occupational activities focusing on exposure to tick bites. A retrospective epidemiologic study evaluating 123,495 electronic patients' records (women, n=61,712; men, n=61,783) and a prospective clinical study including 118 patients (women, n=54; men, n=64) 18 years or older seeking care for EM >or= cm in diameter with genospecies verified by polymerase chain reaction (PCR) were conducted. Women 40 years or older had a 48% higher risk than men 40 years or older and 42% higher risk than women younger than 40 years of attracting tick bites (0.0188 versus 0.0127 and 0.0188 versus 0.0132 tick bites respectively per hour). Additionally they had a 96% higher risk than men younger than 40 years of attracting tick bites (0.0188 versus 0.0096). The annual incidence rate of EM in women was 506 and in men 423 cases per 100,000 inhabitants (p<0.001). Significant differences in incidence rates occurred in those 40 years or older. Odds ratios for males infected with Borrelia afzelii developing nonannular EM were 0.09 (95% confidence interval [CI] 0.03 to 0.33) in comparison to females infected by Borrelia afzeli. Significant gender differences in the risk of contracting tick bites, incidence rates, and clinical picture of EM have been observed. Exposure to tick bites alone may not explain these observations and further studies need to be done to clarify the biologic, immunologic, and sociological mechanisms causing these differences. PMID- 17417956 TI - Detection of Borrelia burgdorferi DNA in lizards from Southern Maryland. AB - Lizards serve as hosts for Ixodes ticks in the western and southeastern United States and may affect the transmission cycles of Borrelia burgdorferi in these regions. In Maryland, the role of lizards in the maintenance and transmission cycle of this pathogen has not been examined. We tested 29 lizards (Sceloporus undulatus and Eumeces spp.) and 21 ticks from these lizards for the presence of B. burgdorferi. Eight lizards were positive by polymerase chain reaction (PCR) for at least one B. burgdorferi-specific marker. This is the first report of B. burgdorferi sensu stricto detected in lizards in the mid-Atlantic region. Although the viability of the B. burgdorferi in these lizards was unconfirmed, recovery of bacterial DNA in a tail snip suggests that the infections in these lizards were disseminated. This study suggests that some lizards indigenous to the mid-Atlantic region may serve as alternative reservoirs for B. burgdorferi. In areas where lizard densities are high, these less efficient reservoirs may affect the enzootic cycle of this tick-borne pathogen. PMID- 17417957 TI - Ixodes ricinus density and infection prevalence of Borrelia burgdorferi sensu lato along a North-facing altitudinal gradient in the Rhone Valley (Switzerland). AB - Questing Ixodes ricinus ticks were sampled monthly along a north-facing altitudinal gradient in the canton of Valais, Switzerland, from March 2004 to February 2005. Tick density and infection with Borrelia burgdorferi sensu lato were monitored. Ticks were collected by flagging vegetation at three different altitudes (750 m, 880 m, and 1020 m above sea level). Ticks were examined for Borrelia by polymerase chain reaction (PCR) followed by reverse line blot. At the three altitudes, questing tick activity was not observed under 10 degrees C and was reduced when saturation deficit was higher than 5 mm Hg, most questing tick activity was occurred between 2 mm Hg and 7 mm Hg. Tick density and peak tick density were highest at 1020 m. High saturation deficits at the lowest altitudes appear to impair the tick population. The prevalence of B. burgdorferi infection in nymphs and adults decreased with altitude. The prevalence of infection was higher in adult ticks (47%) than in nymphs (29%). Four B. burgdorferi sensu lato genospecies were detected: B. afzelii (40%), B. garinii (22%), B. valaisiana (12%) and B. burgdorferi sensu stricto (6%). Mixed infections were detected in 13% of infected ticks. PMID- 17417958 TI - The role of wild boar (Sus scrofa) in the eco-epidemiology of R. slovaca in Northeastern Spain. AB - Rickettsia slovaca is considered the etiological agent of tick-borne lymphadenopathy (TIBOLA), an emerging disease transmitted by tick bites. Dermacentor marginatus constitutes the most important vector and wild boar (Sus scrofa) is the main wild host in our area. The epidemiology of this tick-borne rickettsioses has not been completely clarified. During hunting season 2004, wild boar sera sample were collected from northeastern Spain. Ticks were collected both from wild boar and from flagging vegetation in the same areas where wild boar were hunted. Serologic study was carried out using the immunofluorescent antibody (IFA) technique. Ticks were identified as D. marginatus and Rhipicephalus uranicus. Based on sequence analysis on ompA gene, R. slovaca was identified in 30.5% D. marginatus ticks removed from wild boar and in 33.3% D. marginatus collected from flagging vegetation. Rickettsia sp. RpA4 was identified in 4 specimens of D. marginatus removed from wild boar. Twelve of 23 wild boar were seropositive to R. slovaca. Results suggested wild boar are exposed to R. slovaca infection and this pathogen is well established in the wild cycle of D. marginatus in our area. PMID- 17417959 TI - Prevalence of and risk factors for canine tick infestation in the United States, 2002-2004. AB - Knowledge of the geographic range and seasonal activity of vector ticks is important for determining which people or animals are at risk of acquiring tick borne infections. Several time-consuming methods requiring large-scale organization are used to map geographic and seasonal variations in tick distribution. A cost-effective, complementary approach to study tick distribution using a large nation-wide veterinary database is described in this paper. Prevalence of canine tick infestation in 40 states was estimated by analyzing electronic medical records of more than 8 million dog visits to Banfield veterinary hospitals in 2002-2004. Prevalence was defined as the proportion of dog visits in which tick infestation was recorded, and was expressed per 10,000 dog visits with 95% confidence intervals (CI). The overall prevalence (CI) of tick infestation was 52 (51, 53) dogs per 10,000 dog visits. Among states, Oklahoma (249 [229, 271) dogs with ticks per 10,000 dog-visits), Arkansas (242 [213, 274]), Connecticut (136 [119, 155]), West Virginia (130 [105, 161]), and Rhode Island (122 [97, 154]) ranked highest in prevalence of canine tick infestation. Overall prevalence peaked from May through July, although monthly prevalence varied by geographic region. In multiple logistic regression, younger dogs, male dogs, and sexually intact dogs, were at increased risk of tick infestation. Toy breeds were least likely to be infested, but no linear pattern of risk was evident with body weight. Identified risk factors should enable veterinarians to prevent tick infestation in pet dogs although differences in risk of tick infestation may be related to outdoor activity of dogs. Feasibility of collecting information for surveillance of vectors ticks on a national level using this large, electronic veterinary database is discussed. PMID- 17417960 TI - Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. AB - Aedes albopictus, commonly known as the Asian tiger mosquito, is currently the most invasive mosquito in the world. It is of medical importance due to its aggressive daytime human-biting behavior and ability to vector many viruses, including dengue, LaCrosse, and West Nile. Invasions into new areas of its potential range are often initiated through the transportation of eggs via the international trade in used tires. We use a genetic algorithm, Genetic Algorithm for Rule Set Production (GARP), to determine the ecological niche of Ae. albopictus and predict a global ecological risk map for the continued spread of the species. We combine this analysis with risk due to importation of tires from infested countries and their proximity to countries that have already been invaded to develop a list of countries most at risk for future introductions and establishments. Methods used here have potential for predicting risks of future invasions of vectors or pathogens. PMID- 17417961 TI - Mortality and reproductive dynamics of Aedes aegypti (Diptera: Culicidae) fed human blood. AB - Mortality is a critical factor in determining a mosquito's ability to transmit pathogens. We investigated the effect of human blood feeding and reproduction on mortality of the dengue virus vector, Aedes aegypti, by conducting a life-table study of male and female mosquitoes maintained on one of three diets: 10% sucrose, human blood or human blood plus 10% sucrose. We examined the effect of host availability by offering human blood to mosquitoes every day or every other day. Mortality of females was age-dependent and best fit by a logistic or logistic-Makeham model. The availability of blood increased survival; survival of females fed blood plus sugar was greater than those only fed sugar. There was a peak in mortality of females fed blood alone early in life that coincided with the initiation of oviposition. When females in the blood alone group were offered blood daily, their mortality was significantly lower than when they were offered blood every other day. Unlike some previous studies, females fed blood plus sugar had higher fitness than females fed blood alone. Increased fitness may have been due to differences in housing mosquitoes individually in separate cages versus as a group of many mosquitoes in each cage. It was not due to longer survival of males who had access to sugar as a food source. Our results demonstrate that reproductively active Ae. aegypti exhibit age-dependent mortality, which refutes the assumption of age-independent mosquito mortality and underscores the need to incorporate age-dependent factors into pathogen transmission models and research on mosquito biology in general. PMID- 17417962 TI - Interview with Scott B. Halstead, M.D. PMID- 17417965 TI - Relative scotomata in the "normal" eye of functionally amblyopic patients. A scanning laser ophthalmoscope (SLO) micreperimetric study. AB - PURPOSE: To evaluate amblyopic patients with scanning laser ophthalmoscope (SLO) microperimetry to determine whether SLO assessment and data might provide useful information in our understanding of amblyopia and determine its utility in the evaluation of amblyopic patients. METHODS: In this retrospective case series, selected for SLO testing case series, clinical data of forty-six patients with amblyopia were reviewed after completion of treatment for anisometropic or strabismic amblyopia. Ten ophthalmologically age-matched, normal patients served as controls. All patients were tested with the SLO, evaluating for the presence of macular scotomata. SLO findings were assessed within each group and between groups. RESULTS: A macular scotoma was found in the amblyopic eye of 25 of 26 anisometropic amblyopic patients and all 20 strabismic amblyopia patients. Twenty of 26 patients with anisometropic amblyopia had a relative scotoma in the non amblyopic "normal" eye. All 20 patients with strabismic amblyopia also had a non amblyopic "normal" contralateral eye scotoma. None of the normal control patients had a scotoma in either eye. Several ocular and binocular clinical features were correlated to scotoma findings within and between groups. CONCLUSION: The SLO proved useful for the assessment of some features of amblyopia. A scotoma was identified not only in the amblyopic eye of all but one amblyopic patient, as expected, but also in almost all of the fellow non- amblyopic, presumed "normal" contralateral eyes, and in spite of treatment normalization of visual acuity and stereoacuity in several cases. Thus, the ocular and binocular pathological effects of unilaterally functional amblyopia are not limited to the amblyopic eye but may also be seen, to a sub-clinical degree, by SLO microperimetry in the supposedly normal contralateral eye as well as in the apparently successfully treated previously amblyopic eye. PMID- 17417966 TI - Long term vision outcomes of conventional treatment of strabismic and anisometropic functional amblyopia. AB - PURPOSE: To investigate the long-term vision outcomes of amblyopia treatment in "successfully" compared with "unsuccessfully" treated patients. METHODS: Forty two participants (n=42, mean age 14.8 years, range 10-25 years) were enrolled in the study. Individuals with strabismic or mixed (strabismic and anisometropic) amblyopia were examined at a mean of 6.6 years (range 1-18 years) after cessation of amblyopia treatment. Participants were classified as being "successfully" treated (Group 1) if visual acuity of 6/7.5 or better was achieved at cessation of treatment, or "unsuccessfully" treated (Group 2) if visual acuity of 6/9 or less was achieved at cessation of treatment. Visual acuity was analyzed by calculating an interocular score or difference in visual acuity between the amblyopic and non amblyopic normal (control) eye. RESULTS: A deterioration of visual acuity occurred in 62% of the participants in both Groups 1 and 2. The mean deterioration of visual acuity over time for either group was less than one LogMAR chart line and was not "statistically significant" by convention (F [1,39]=3.361, p=0.074). The outcomes achieved at cessation of treatment did not "statistically significantly" affect the mean deterioration that occurred over time (F [1,49]=0.031, p=0.860). CONCLUSION: Visual acuity was relatively stable over a mean followup period of 6.6 years. The treatment outcome and the success of amblyopia treatment were found to be irrelevant to long term stability of visual acuity. These findings suggest that amblyopia treatment mostly results in a lasting improvement in visual acuity, and that both unsuccessfully and successfully treated individuals maintain their visual acuity improvement achieved during treatment. PMID- 17417967 TI - Efficient production and secretion of bovine beta-lactoglobulin by Lactobacillus casei. AB - BACKGROUND: Lactic acid bacteria (LAB) are attractive tools to deliver therapeutic molecules at the mucosal level. The model LAB Lactococcus lactis has been intensively used to produce and deliver such heterologous proteins. However, compared to recombinant lactococci, lactobacilli offer some advantages such as better survival in the digestive tract and immunomodulatory properties. Here, we compared different strategies to optimize the production of bovine beta lactoglobulin (BLG), a major cow's milk allergen, in the probiotic strain Lactobacillus casei BL23. RESULTS: Using a nisin-inducible plasmid system, we first showed that L. casei BL23 strain could efficiently secrete a reporter protein, the staphylococcal nuclease (Nuc), with the lactococcal signal peptide SPUsp45 fused to its N-terminus. The fusion of SPUsp45 failed to drive BLG secretion but led to a 10-fold increase of intracellular BLG production. Secretion was significantly improved when the synthetic propeptide LEISSTCDA (hereafter called LEISS) was added to the N-terminus of the mature moiety of BLG. Secretion rate of LEISS-BLG was 6-fold higher than that of BLG alone while intracellular production reached then about 1 mg/L of culture. The highest yield of secretion was obtained by using Nuc as carrier protein. Insertion of Nuc between LEISS and BLG resulted in a 20-fold increase in BLG secretion, up to 27 microg/L of culture. Furthermore, the lactococcal nisRK regulatory genes were integrated into the BL23 chromosome. The nisRK insertion allowed a decrease of BLG synthesis in uninduced cultures while BLG production increased by 50% after nisin induction. Moreover, modification of the induction protocol led to increase the proportion of soluble BLG to around 74% of the total BLG production. CONCLUSION: BLG production and secretion in L. casei were significantly improved by fusions to a propeptide enhancer and a carrier protein. The resulting recombinant strains will be further tested for their ability to modulate the immune response against BLG via mucosal delivery in a cow's milk allergy model in mice. PMID- 17417968 TI - Identification of typical medullary breast carcinoma as a genomic sub-group of basal-like carcinomas, a heterogeneous new molecular entity. AB - INTRODUCTION: Typical medullary breast carcinoma (MBC) has recently been recognized to be part of the basal-like carcinoma spectrum, a feature in agreement with the high rate of TP53 mutations previously reported in MBCs. The present study was therefore designed to identify phenotypic and genetic alterations that distinguish MBCs from basal-like carcinomas (BLC). METHODS: Expression levels of estrogen receptor (ER), progesterone receptor (PR), ERBB2, TP53, cytokeratins (KRTs) 5/6, 14, 8/18, epidermal growth factor receptor and KIT, as well as TP53 gene sequence and high-density array comparative genomic hybridization (CGH) profiles, were assessed and compared in a series of 33 MBCs and 26 BLCs. RESULTS: All tumors were negative for ER, PR and ERBB2. KRTs 5/6 were more frequently expressed in MBCs (94%) than in BLCs (56%) (p = 0.0004). TP53 mutations were disclosed in 20/26 MBCs (77%) and 20/24 BLCs (83%). Array CGH analysis showed that a higher number of gains (95 regions) and losses (34 regions) was observed in MBCs than in BLCs (36 regions of gain; 13 regions of losses). In addition, gains of 1q and 8q, and losses of X were found to be common to the two groups, whereas gains of 10p (53% of the cases), 9p (30.8% of the cases) and 16q (25.8% of the cases), and losses of 4p (34.8% of the cases), and amplicons of 1q, 8p, 10p and 12p were the genetic alterations found to characterize MBC. CONCLUSION: Our study has revealed that MBCs are part of the basal-like group and share common genomic alterations with BLCs, the most frequent being 1q and 8q gains and X losses; however, MBCs are a distinct entity within the basal-like spectrum, characterized by a higher rate of KRT 5/6 expression, a higher rate of gains and losses than BLCs, recurrent 10p, 9p and 16q gains, 4p losses, and 1q, 8p, 10p and 12p amplicons. Our results thus contribute to a better understanding of the heterogeneity in basal-like breast tumors and provide potential diagnostic tools. PMID- 17417969 TI - Large-scale RNAi screens identify novel genes that interact with the C. elegans retinoblastoma pathway as well as splicing-related components with synMuv B activity. AB - BACKGROUND: The retinoblastoma tumor suppressor (Rb) acts in a conserved pathway that is deregulated in most human cancers. Inactivation of the single Rb-related gene in Caenorhabditis elegans, lin-35, has only limited effects on viability and fertility, yet causes changes in cell-fate and cell-cycle regulation when combined with inactivation of specific other genes. For instance, lin-35 Rb is a synthetic multivulva (synMuv) class B gene, which causes a multivulva phenotype when inactivated simultaneously with a class A or C synMuv gene. RESULTS: We used the ORFeome RNAi library to identify genes that interact with C. elegans lin-35 Rb and identified 57 genes that showed synthetic or enhanced RNAi phenotypes in lin-35 mutants as compared to rrf-3 and eri-1 RNAi hypersensitive mutants. Based on characterizations of a deletion allele, the synthetic lin-35 interactor zfp-2 was found to suppress RNAi and to cooperate with lin-35 Rb in somatic gonad development. Interestingly, ten splicing-related genes were found to function similar to lin-35 Rb, as synMuv B genes that prevent inappropriate vulval induction. Partial inactivation of specific spliceosome components revealed further similarities with lin-35 Rb functions in cell-cycle control, transgene expression and restricted expression of germline granules. CONCLUSION: We identified an extensive series of candidate lin-35 Rb interacting genes and validated zfp-2 as a novel lin-35 synthetic lethal gene. In addition, we observed a novel role for a subset of splicing components in lin-35 Rb-controlled processes. Our data support novel hypotheses about possibilities for anti-cancer therapies and multilevel regulation of gene expression. PMID- 17417970 TI - Takotsubo cardiomyopathy in a Caucasian Italian woman: case report. AB - BACKGROUND: Takotsubo cardiomyopathy is an acute cardiac syndrome characterized by transient LV regional wall motion abnormalities (with peculiar apical ballooning appearance), chest pain or dyspnea, ST-segment elevation and minor elevations of cardiac enzyme levels CASE PRESENTATION: A 68-year-old woman was admitted to the Emergency Department because of sudden onset chest pain occurred while transferring her daughter, who had earlier suffered a major seizure, to the hospital. The EKG showed sinus tachycardia with ST-segment elevation in leads V2 V3 and ST-segment depression in leads V5-V6, she was, thus, referred for emergency coronary angiography. A pre-procedural transthoracic echocardiogram revealed regional systolic dysfunction of the LV walls with hypokinesis of the mid-apical segments and hyperkinesis of the basal segments. Coronary angiography showed patent epicardial coronary arteries; LV angiography demonstrated the characteristic morphology of apical ballooning with hyperkinesis of the basal segments and hypokinesis of the mid-apical segments. The post-procedural course was uneventful; on day 5 after admission the echocardiogram revealed full recovery of apical and mid-ventricular regional wall-motion abnormalities. CONCLUSION: Takotsubo cardiomyopathy is a relatively rare, unique entity that has only recently been widely appreciated. Acute stress has been indicated as a common trigger for the transient LV apical ballooning syndrome, especially in postmenopausal women. The present report is a typical example of stress-induced takotsubo cardiomyopathy in a Caucasian Italian postmenopausal woman. PMID- 17417972 TI - Genetic affinities between endogamous and inbreeding populations of Uttar Pradesh. AB - BACKGROUND: India has experienced several waves of migration since the Middle Paleolithic. It is believed that the initial demic movement into India was from Africa along the southern coastal route, approximately 60,000-85,000 years before present (ybp). It has also been reported that there were two other major colonization which included eastward diffusion of Neolithic farmers (Elamo Dravidians) from Middle East sometime between 10,000 and 7,000 ybp and a southern dispersal of Indo Europeans from Central Asia 3,000 ybp. Mongol entry during the thirteenth century A.D. as well as some possible minor incursions from South China 50,000 to 60,000 ybp may have also contributed to cultural, linguistic and genetic diversity in India. Therefore, the genetic affinity and relationship of Indians with other world populations and also within India are often contested. In the present study, we have attempted to offer a fresh and immaculate interpretation on the genetic relationships of different North Indian populations with other Indian and world populations. RESULTS: We have first genotyped 20 tetra-nucleotide STR markers among 1800 north Indian samples of nine endogamous populations belonging to three different socio-cultural strata. Genetic distances (Nei's DA and Reynold's Fst) were calculated among the nine studied populations, Caucasians and East Asians. This analysis was based upon the allelic profile of 20 STR markers to assess the genetic similarity and differences of the north Indian populations. North Indians showed a stronger genetic relationship with the Europeans (DA 0.0341 and Fst 0.0119) as compared to the Asians (DA 0.1694 and Fst - 0.0718). The upper caste Brahmins and Muslims were closest to Caucasians while middle caste populations were closer to Asians. Finally, three phylogenetic assessments based on two different NJ and ML phylogenetic methods and PC plot analysis were carried out using the same panel of 20 STR markers and 20 geo ethnic populations. The three phylogenetic assessments revealed that north Indians are clustering with Caucasians. CONCLUSION: The genetic affinities of Indians and that of different caste groups towards Caucasians or East Asians is distributed in a cline where geographically north Indians and both upper caste and Muslim populations are genetically closer to the Caucasians. PMID- 17417971 TI - Sequential emergence and clinical implications of viral mutants with K70E and K65R mutation in reverse transcriptase during prolonged tenofovir monotherapy in rhesus macaques with chronic RT-SHIV infection. AB - BACKGROUND: We reported previously on the emergence and clinical implications of simian immunodeficiency virus (SIVmac251) mutants with a K65R mutation in reverse transcriptase (RT), and the role of CD8+ cell-mediated immune responses in suppressing viremia during tenofovir therapy. Because of significant sequence differences between SIV and HIV-1 RT that affect drug susceptibilities and mutational patterns, it is unclear to what extent findings with SIV can be extrapolated to HIV-1 RT. Accordingly, to model HIV-1 RT responses, 12 macaques were inoculated with RT-SHIV, a chimeric SIV containing HIV-1 RT, and started on prolonged tenofovir therapy 5 months later. RESULTS: The early virologic response to tenofovir correlated with baseline viral RNA levels and expression of the MHC class I allele Mamu-A*01. For all animals, sensitive real-time PCR assays detected the transient emergence of K70E RT mutants within 4 weeks of therapy, which were then replaced by K65R mutants within 12 weeks of therapy. For most animals, the occurrence of these mutations preceded a partial rebound of plasma viremia to levels that remained on average 10-fold below baseline values. One animal eventually suppressed K65R viremia to undetectable levels for more than 4 years; sequential experiments using CD8+ cell depletion and tenofovir interruption demonstrated that both CD8+ cells and continued tenofovir therapy were required for sustained suppression of viremia. CONCLUSION: This is the first evidence that tenofovir therapy can select directly for K70E viral mutants in vivo. The observations on the clinical implications of the K65R RT-SHIV mutants were consistent with those of SIVmac251, and suggest that for persons infected with K65R HIV-1 both immune-mediated and drug-dependent antiviral activities play a role in controlling viremia. These findings suggest also that even in the presence of K65R virus, continuation of tenofovir treatment as part of HAART may be beneficial, particularly when assisted by antiviral immune responses. PMID- 17417973 TI - Serum antioxidant micromineral (Cu, Zn, Fe) status of drug dependent subjects: Influence of illicit drugs and lifestyle. AB - BACKGROUND: Use of illicit drugs induces multiple nutrient deficiencies. Drug habit, sexual practice and socioeconomic factors influence the nutrient profile of drug dependent subjects. The literature on this issue is still insufficient. This study has tested the hypothesis that illicit drug use and lifestyle impair mineral status. To test this hypothesis, 253 men multiple drug users of age 18-45 years were recruited to investigate their serum copper, zinc and iron levels. Influence of illicit drugs and their lifestyle on the mineral levels was also examined. The study subjects were drug dependent who had shared needles and had sexual activity with multiple partners. Serum concentrations of the minerals were estimated by atomic absorption flame spectrometry. RESULTS: Results showed a significant increase in serum copper and zinc concentrations, and decrease in iron level in drug dependent subjects. The increase of copper level was found to be much higher than that of zinc. Period of drug abuse had made a significant positive influence on the copper and iron levels, but it was apparently reversed for zinc concentration. Multiple sexual partnerships had significant influence on zinc status. There also were significant relationships observed between body mass index (BMI) as well as certain socioeconomic factors, and mineral status of drug dependent subjects and non-drug dependent controls. A series of multiple linear regression analysis predicted mineral values for education, age and BMI. The group (drug dependent subject = 1, non-drug dependent control = 2) had a significant influence on these parameters. However, after controlling these factors, it was shown that illicit drug use significantly contributed to influence the serum mineral levels. CONCLUSION: Illicit drug use impairs serum mineral value causing an increase in copper and zinc and a decrease in iron. Lifestyle and nutritional status of drug dependent subjects influence serum mineral concentrations. PMID- 17417974 TI - Respiratory controversies in the critical care setting. Part I. PMID- 17417975 TI - Respiratory controversies in the critical care setting. When caring for critically ill patients, do clinicians have a responsibility to be innovative and try unproven approaches when accepted approaches are failing? AB - As the first paper in this Journal Conference on intensive care unit controversies, the editors wished us to set the tone for the debate by discussing the ethics of medical "adventurism" in the intensive care unit. More life-or death decisions are made in the intensive care unit than elsewhere in the hospital, and the critical care specialist often sees himself or herself as a warrior in a battle with death. This adrenaline-charged calling attracts highly intelligent, hard-working, and compassionate caregivers, as well as fiercely independent clinicians. The result of this is that critical care specialists passionately debate about the meaning and application of published "evidence" and this leads to thoughtful debate, as exemplified by the papers in this and the next issue of Respiratory Care, as well as thoughtless and often dangerous disregard for evidence-based medicine. Physicians are morally obligated to provide the best and most appropriate care possible for their patients, but when accepted approaches are failing and a critically ill patient is getting worse, the critical care physician must make a decision regarding innovative therapy, based on the patient's prognosis, the available evidence, the resources on hand, the expertise of the physicians, and the values of the patient and the physician. This decision may lead, at times, to trying unproven and innovative strategies to achieve a clinical goal. In such cases, it is to be hoped that this can be done in such a way that data are formally and prospectively collected to increase our knowledge. PMID- 17417976 TI - Respiratory therapies in the critical care setting. Should aerosolized antibiotics be administered to prevent or treat ventilator-associated pneumonia in patients who do not have cystic fibrosis? AB - Ventilator-associated pneumonia (VAP) significantly increases intensive care unit morbidity, mortality, and costs. VAP is thought to be caused by bacterial entry into injured airways, which produces tracheobronchitis that evolves into diffuse pneumonia. The use of aerosolized antibiotics is conceptually attractive, especially when the infection is early and limited to the airway epithelium. Data show that aerosolized antibiotics kill airway bacteria and improve outcomes in cystic fibrosis. The clinical evidence for aerosolized antibiotics to prevent VAP is weak but suggestive. Concerns about the high cost, possible development of antibiotic resistance, and other potential risks of aerosolized antibiotics led several evidence-based consensus groups to recommend against routine use of aerosolized antibiotics for VAP prevention until better data are available. Importantly, the clinical evidence that aerosolized antibiotics can treat established VAP is negative, and multiple consensus groups recommend against treating established VAP with aerosolized antibiotics. PMID- 17417977 TI - Respiratory therapies in the critical care setting. Should every mechanically ventilated patient be monitored with capnography from intubation to extubation? AB - One of the most important aspects of caring for a critically ill patient is monitoring. Few would disagree that the most essential aspect of monitoring is frequent physical assessments. Complementing the physical examination is continuous monitoring of heart rate, respiratory rate, and blood oxygen saturation measured via pulse-oximetry, which have become the standard of care in intensive care units. Over the past decade one of the most controversial aspects of monitoring critically ill patients has been capnography. Although most clinicians use capnography to confirm endotracheal intubation, few clinicians use continuous capnography in the intensive care unit. This article reviews the medical literature on whether every mechanically ventilated patient should be monitored with capnography from intubation to extubation. There are numerous articles on capnography, but no definitive, randomized study has even attempted to address this specific question. Based on the available literature, it seems reasonable to use continuous capnography, for at least a subset of critically ill patients, to ensure integrity of the endotracheal tube and other ventilatory apparatus. However, at this point definitive data are not yet available to clearly support continuous capnography for optimizing mechanical ventilatory support. We hope that as new data become available, the answer to this capnography question will become clear. PMID- 17417978 TI - Respiratory controversies in the critical care setting. Should all patients be treated with hypothermia following cardiac arrest? AB - Cardiac arrest is a common and lethal medical problem; each year more than half a million people in the United States and Canada suffer cardiac arrest treated by emergency medical personnel or in-hospital providers. Of those who survive to hospital admission or suffer in-hospital arrest, 40-60% die prior to discharge. Neurologic injury is the major source of morbidity and mortality after recovery of spontaneous circulation. Therapeutic options to prevent neurologic injury are limited, but recent randomized trials showed that moderate therapeutic hypothermia improves neurologic outcome in selected patients following cardiac arrest. Clear consensus statements recommend that unconscious adult patients with spontaneous circulation after out-of-hospital cardiac arrest should be cooled if the initial rhythm was ventricular fibrillation, and that therapeutic hypothermia should be considered for other patients (other rhythms or in-hospital arrest). However, the position that all patients should be cooled following cardiac arrest is probably too broad, given the lack of studies on patients with non-ventricular fibrillation rhythms, in-hospital arrest, or non-cardiac causes of arrest. Further research is needed to determine the broadest application of moderate therapeutic hypothermia. PMID- 17417979 TI - Respiratory controversies in the critical care setting. Does airway pressure release ventilation offer important new advantages in mechanical ventilator support? AB - Airway pressure-release ventilation (APRV) is a mechanical ventilation strategy that is usually time-triggered but can be patient-triggered, pressure-limited, and time-cycled. APRV provides 2 levels of airway pressure (P(high) and P(low)) during 2 time periods (T(high) and T(low)), both set by the clinician. APRV usually involves a long T(high) and a short T(low). APRV uses an active exhalation valve that allows spontaneous breathing during both T(high) and T(low). APRV typically generates a higher mean airway pressure with a lower tidal volume (V(T)) and lower positive end-expiratory pressure than comparable levels of other ventilation strategies, so APRV may provide better alveolar recruitment at a lower end-inflation pressure and therefore (1) decrease the risk of barotrauma and alveolar damage in patients with acute lung injury or acute respiratory distress syndrome (ALI/ARDS), and (2) provide better ventilation perfusion matching, cardiac filling, and patient comfort than modes that do not allow spontaneous breaths. However, if the patient makes a spontaneous breath during T(high), the V(T) generated could be much larger than the clinician-set target V(T), which could cause the end-inflation transpulmonary pressure and alveolar stretch to be much larger than intended or produced in other ventilation strategies. It is unknown whether a patient's inspiratory effort (and consequent larger V(T)) can damage alveoli in the way that mechanically delivered, positive pressure breaths can damage alveoli in ALI/ARDS. Other ventilation modes also promote spontaneous breaths, but at overall lower end-inflation transpulmonary pressure. There is a dearth of data on what would be the optimal APRV inspiratory expiratory ratio, positive end-expiratory pressure, or weaning strategy. The few clinical trials to date indicate that APRV provides adequate gas exchange, but none of the data indicate that APRV confers better clinical outcomes than other ventilation strategies. PMID- 17417980 TI - Respiratory controversies in the critical care setting. Do the NIH ARDS Clinical Trials Network PEEP/FIO2 tables provide the best evidence-based guide to balancing PEEP and FIO2 settings in adults? AB - Positive end-expiratory pressure (PEEP) and inspired oxygen fraction (F(IO(2))) are the primary means of improving P(aO(2)) during mechanical ventilation. Patients with acute respiratory distress syndrome (ARDS) typically present with a large intrapulmonary shunt, which makes even high F(IO(2)) ineffective in improving P(aO(2)). PEEP decreases intrapulmonary shunt by recruiting collapsed alveoli, but PEEP is associated with important adverse effects, whereas prolonged exposure to high F(IO(2)) may cause oxidative lung injury. The improved survival found in the National Institutes of Health's ARDS Network low-tidal-volume study may suggest that their PEEP/F(IO(2)) titration tables represent the best method for adjusting these variables. Based upon an extensive literature review of PEEP and respiratory system mechanics in ARDS, we conclude that: (1) for most patients the therapeutic range of PEEP is relatively narrow, so the ARDS Network PEEP/F(IO(2)) strategy is reasonable and supported by high-level evidence, (2) how best to adjust PEEP to prevent or ameliorate ventilator-associated lung injury is unknown and still under investigation, and (3) in a small subset of patients with severe lung injury and/or abnormal chest-wall compliance, highly individualized titration of PEEP, based upon the respiratory-system pressure volume curve, PEEP/tidal-volume titration grids, or a recruitment maneuver and a PEEP decrement trial is a reasonable alternative. PMID- 17417981 TI - Controversies in the critical care setting. Should adaptive pressure control modes be utilized for virtually all patients receiving mechanical ventilation? AB - Traditional mechanical ventilation is provided with either a constant volume or constant pressure breath. In recent years, dual-control (adaptive pressure control) has been introduced in an attempt to combine the attributes of volume ventilation (constant tidal volume and minute ventilation) with the attributes of pressure ventilation (rapid, variable flow and reduced work of breathing). Adaptive pressure control is a pressure-controlled breath that utilizes closed loop control of the pressure setting to maintain a minimum delivered tidal volume. Prior to the introduction of adaptive pressure control, no clinical studies were accomplished. Studies have shown that adaptive pressure control reduces peak inspiratory pressure, compared to volume control. When compared to traditional pressure-control ventilation, no differences have been identified. While adaptive pressure control can guarantee a minimum tidal volume, it cannot guarantee a constant tidal volume. One concern is that the ventilator cannot distinguish between improved pulmonary compliance and increased patient effort. Clinicians should be aware of the limitations of adaptive pressure control and understand when other breath delivery techniques are more suitable. PMID- 17417982 TI - Epidemiology of Sjogren's syndrome: where are we now? PMID- 17417983 TI - Ultrasound imaging for the rheumatologist VII. Ultrasound imaging in rheumatoid arthritis. AB - The present review provides an update of the available data and discusses research issues of ultrasound (US) imaging in rheumatoid arthritis (RA). Currently the principal indications for using US in the assessment of patients with RA include: detection of sub-clinical synovitis, demonstration of bone erosion undetected by conventional radiography, detailed assessment of tendon pathology and guided injection and aspiration of joints and soft tissues. Future potential applications are likely to include short and long term therapy monitoring and early detection of cartilaginous changes in RA. The main priorities requiring the attention of investigators include: addressing validity issues, especially those related to criterion and discriminator validity, development of international consensus on scoring systems, evaluation of the role of power Doppler in the assessment of disease activity, development of a specific training programme for rheumatologists performing US and investigation of the potential of 3D US using a volumetric probe. PMID- 17417984 TI - Effects of celecoxib on human chondrocytes--enhanced production of chemokines. AB - OBJECTIVE: The purpose of this study was to examine the effects of a selective cyclooxigenase-2 (COX-2) inhibitor (celecoxib) comparing diclofenac. METHODS: Using chondrocytes derived from cartilage of non-arthritic (NA) subjects or patients with osteoarthritis (OA) or rheumatoid arthritis (RA), we examined the effects of celecoxib on incorporation of 3H-thymidine and 35S-sulfate, apoptosis, and production of matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, and regulated upon activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein (MIP)-1a and nitric oxide (NO). RESULTS: Celecoxib and diclofenac tended to reduce 3H-tymidine incorporation of chondrocytes. Celecoxib induced apoptosis in a dose-dependent manner, but to a lesser degree than diclofenac. Celecoxib inhibited proteoglycan synthesis (indicated by 35S-sulfate incorporation) in NA chondrocytes, but not in OA and RA chondrocytes. Celecoxib increased interleukin-1 (IL-1)-induced production of RANTES and MIP-1alpha by chondrocytes and decreased IL-1-induced NO production by chondrocytes, whereas it did not affect MMP production. CONCLUSION: Celecoxib had both beneficial and adverse effects on chondrocytes. RA, OA and NA chondrocytes showed different responses. Interestingly, celecoxib enhanced the production of chemokines. PMID- 17417985 TI - Cell clonality in hypereosinophilic syndrome: what pathogenetic role? AB - OBJECTIVE: Idiopathic hypereosinophilic syndrome (HES) is a heterogeneous disorder, including either a myeloproliferative or a lymphoproliferative variant (l-HES). In l-HES, T-lymphocytes could be involved in the pathogenesis through several cytokines, including IL5. METHODS: We assayed both TCR Beta- and delta rearrangements by fluorescent PCR, characterizing 14 patients affected by HES. Lyn activation (a src-kinase involved in the IL5 pathway) was also tested in 6 cases. RESULTS: FIP1L1-PDGFRa was detected in 4 cases (28.6%); a clonal TCR was found in 10 cases (71.4%), including cases FIP1L1-PDGFRalpha-positive; four cases did not show any molecular marker. In this series, levels of IL5, IL4, IL2 and gammaIFN were measured, without any significant difference among different subgroups. All pathological samples tested did not show Lyn activation. Immunophenotype was also characterized: only one case showed an atypical CD3 /CD4+ population in the bone marrow. CONCLUSION: This study would suggest that a real distinction between m- and l-HES is not wholly convincing and that clonal T cell expansion could not be the "primum movens" but an epiphenomenon in HES. PMID- 17417986 TI - Infliximab therapy in pulmonary fibrosis associated with collagen vascular disease. AB - OBJECTIVE: To study the potential effectiveness of tumor necrosis factor a (TNF alpha) inhibitor treatment for pulmonary fibrosis associated with a collagen vascular disease, CVD (rheumatoid arthritis, RA and systemic sclerosis, SSc) refractory to conventional treatment. METHODS: Four patients (three men with RA, one woman with SSc) were treated with infliximab. All patients received 3mg/kgr of infliximab at intervals 0, 2 and 6 weeks, and then maintenance infusions every 8 weeks afterwards for at least a 12-month period. Patients had active disease despite treatment with corticosteroids and other immunomodulatory agents. RESULTS: Treatment was well-tolerated from all patients. Pulmonary fibrosis remained stable during treatment in terms of symptoms, pulmonary function tests (PFTs) and High resolution computed tomography (HRCT) appearance. As expected, a clinical response was observed in joint symptoms in patients with RA as evaluated by the DAS28 (Disease Activity Score, the 28 joint version). CONCLUSION: This study suggests that inhibition of TNF-alpha with infliximab may stabilize the progression of pulmonary fibrosis associated with CVD. Prospective, controlled trials are necessary to determine the efficacy of infliximab in pulmonary fibrosis associated CVD. PMID- 17417987 TI - Oxaceprol--a randomised, placebo-controlled clinical study in osteoarthritis with a non-conventional non-steroidal anti-inflammatory drug. AB - OBJECTIVE: To evaluate efficacy of therapy with oxaceprol in the treatment of symptomatic osteoarthritis of knee or hip. METHODS: A 3-week prospective, multicentric, randomised, double-blind, placebo-controlled study with 167 patients aged between 40 and 75 years with painful and radiologically confirmed knee or hip osteoarthritis. Patients were randomly assigned to receive oxaceprol 1200 mg/day or placebo for 3 weeks. At inclusion, osteoarthritis symptoms were minimum pain following exercise (standardised as pain after climbing 12-15 stairs) of 40 to 90 mm on a 100 mm pain scale and difficulties in climbing stairs. Efficacy criteria were changes in pain shown in a visual analogue scale (VAS), in the Lequesne index, and in assessments of joint limitation, joint complaint and therapeutic success. The primary end point was the pain following exercise. The confirmatory analysis was based on the Full Analysis data set using the t-test for independent samples. RESULTS: Baseline characteristics of both groups were comparable. In the primary endpoint a clinically relevant and statistically significant superiority of oxaceprol as compared to placebo could be demonstrated (mean improvement in pain following exercise was 16.6 mm in the oxaceprol and 4.5 mm in the placebo group, p = 0.002). The safety and tolerability was good, showing no statistically significant difference between oxaceprol and placebo. CONCLUSION: A statistically significant and clinically relevant efficacy of oxaceprol was shown. The good safety and tolerability of oxaceprol was confirmed. PMID- 17417988 TI - Rheumatic manifestations of lymphoproliferative disorders. AB - OBJECTIVE: The association between musculoskeletal features and lymphoproliferative disorders as well as the association of rheumatic diseases with an increased risk of malignancies is well-known. METHODS: This paper describes three patients with inflammatory joint diseases treated with disease modifying antirheumatic drugs who developed lymphoproliferative disorders either of an abnormal type or with unusual clinical features. RESULTS: The difficulty in differentiating the musculoskeletal symptoms of an underlying lymphoproliferative disease from the features of the rheumatic disease itself with special regard to treatment with disease-modifying antirheumatic drugs including biologics is emphasised on the example of patient 1. Patient 2 presented with a rare type of lymphoma and had been mistakenly diagnosed as having seronegative RA. The last patient with oligoarthritis represents an example of the sarcoidosis-lymphoma syndrome. CONCLUSION: This article addresses several of the problems rheumatologists may experience with the various rheumatologic manifestations of lymphoproliferative disorders. Until more definitive data are available, patients who develop unexpected arthritis should be considered for histologic biopsy to rule out coexistent neoplasia. PMID- 17417989 TI - The immunogenicity, safety, and efficacy of etanercept liquid administered once weekly in patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the immunogenicity, safety, and efficacy of 50 mg/mL liquid etanercept. METHODS: In a multicenter, open-label study, adults with active rheumatoid arthritis (RA) received 50 mg/mL liquid etanercept subcutaneously once weekly for 24 weeks. Immunogenicity was assessed at baseline and weeks 24 and 28, safety at all study visits, and efficacy at baseline and weeks 12 and 24. RESULTS: Of 222 treated patients, 88% completed the study; 81% were women; 84% were white; mean age was 53 years; mean RA duration was 10 years. Antibodies to etanercept, all non-neutralizing, were detected in 12 of 214 patients; 7 of the 12 were borderline positive (antibody titers <1:50). The presence of non-neutralizing anti-etanercept antibodies did not appear to affect clinical safety or efficacy. Few patients reported serious adverse events (6.3%), serious infections (2.3%), or withdrew because of adverse events (4.5%). Most adverse events were mild or moderate. The most common event, injection site reaction, occurred in 29.3% patients. At week 24, 63% of patients achieved an ACR20 response, 36% an ACR50 response, and 14% an ACR70 response. Similar responses were apparent by week 12. Week 24 mean improvement in the Health Assessment Questionnaire disability index scores was 0.6 points; improvement in the Short Form-36 Physical Component Score was 10.0 points. CONCLUSION: The 50 mg/mL liquid etanercept formulation administered once weekly was well tolerated. The incidence of anti-etanercept antibodies, the nature and frequency of adverse events, and improvements in signs and symptoms of RA and patient physical function were similar to those in previous etanercept studies. PMID- 17417990 TI - Erosive arthropathy: clinical variance in lupus erythematosus and association with anti-CCP case series and review of the literature. AB - OBJECTIVE: To describe the occurrence of erosive arthropathy in systemic lupus erythematosus (SLE) and its relationship to anti-CCP antibodies. METHODS: Retrospective medical record review of a case series of five female patients with SLE and erosive arthropathies. RESULTS: The initial disease presentation in all patients was a polyarthritis. Anti-CCP antibodies were detected in 4 out of 5 (80%) patients, 2 of whom had a positive rheumatoid factor. CONCLUSION: Erosive arthritis was strongly associated with the presence of anti-CCP antibodies in these patients with SLE, who presented with polyarthritis. Anti-CCP in patients with SLE may be a marker of a more severe joint disease. PMID- 17417991 TI - A low prevalance of purified protein derivative test positivity in Turkish patients with rheumatoid arthritis. Association with clinical features and HRCT findings. AB - OBJECTIVES: In this study, we aimed to evaluate the frequency of purified protein derivative (PPD) skin test positivity and associated clinical features in RA patients. MATERIALS AND METHODS: We included 94 (80 F, 14 M, mean age: 55.8) consecutive RA patients with a disease duration of 8.7 years. PPD test was performed in all RA patients; clinical features were recorded down; chest x-ray, pulmonary function tests and HRCT were available in all cases. As the control group, we included data of 21 SLE, 44 AS, 27 OA, 16 gouty arthritis and 18 vasculitis patients. RESULTS: The frequencies of PPD positivity in RA (29.8%) and SLE (19%) patients were lower than in patients with AS (65.9%), gouty arthritis (68.8%) and OA (63%) (all p values < 0.01). PPD-positive RA patients were more frequently smokers (p = 0.005) and had a higher rate of RF seropositivity (p = 0.04) than PPD-negatives. PPD was less frequently positive in erosive RA disease (p = 0.033). Chest x-rays and HRCT abnormalities were detected in 41.8% and 62.7% of RA patients, respectively. Frequencies of chest x-ray and HRCT abnormalities in PPD-positive and PPD-negative patients were not different from each other (p > 0.05). CONCLUSION: In our country in which tuberculosis is relatively frequent contrary to the situation in AS patients- we observed a lower frequency of PPD positivity in RA and SLE patients compared to patients with other rheumatic diseases. We did not find any relationship between PPD positivity and the frequency of chest x-ray, HRCT abnormalities. PMID- 17417992 TI - Levels of circulating endothelial progenitor cells in systemic sclerosis. AB - OBJECTIVE: Contradictory results have been reported regarding vasculogenesis in systemic sclerosis (SSc). Our aim was to investigate bone marrow-derived circulating endothelial precursors (EPCs) and activated circulating endothelial cells (CECs) in SSc patients. METHODS: Peripheral blood from consecutive patients with SSc hospitalised for systemic follow-up was analysed and compared with blood from patients with active refractory rheumatoid arthritis (RA) and osteoarthritis (OA). EPCs were quantified by cell sorting and flow cytometry and were identified as circulating CD34+CD133+ cells. Activated CECs were defined as CD105+CD62+CD105+CD102+CD105+CD106+ cells. RESULTS: Patients with SSc had higher putative EPC levels than OA patients, but lower levels than RA patients. In SSc patients, EPC levels increased with European disease activity score. Activated CEC levels were high in SSc patients and RA patients, but not correlated with EPC levels. CONCLUSION: These results together and previous data suggest that EPCs may be recruited during active vascular disease but that the sustained ischaemic conditions of SSc may eventually lead to EPCs depletion. PMID- 17417993 TI - Radiological scoring methods for ankylosing spondylitis: a comparison between the Bath Ankylosing Spondylitis Radiology Index and the modified Stoke Ankylosing Spondylitis Spine Score. AB - OBJECTIVE: The main objective of the present study was to test the interobserver reliability, truth, discrimination and feasibility of two scoring methods available in ankylosing spondylitis (AS) over a follow-up period of 3 years. METHODS: Two blinded trained observers scored 95 AS radiographs from a cohort of AS patients. Each radiograph was scored by two scoring methods, the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS), and the Bath Ankylosing Spondylitis Radiology Index--spine (BASRI-spine). Interobserver agreement was analyzed by intraclass correlation coefficients (ICC). The construct validity was assessed by examining the correlation of the scoring methods with measures of spinal mobility (Bath Ankylosing Spondylitis Metronomy Index--BASMI), functional limitation (Bath Ankylosing Spondylitis Functional Index--BASFI) and disease duration. Bland and Altman's 95% limits of agreement method and effect size (ES) analysis were used to estimate the smallest detectable difference (SDD) of radiological progression and responsiveness. RESULTS: The BASRI-spine reached intra- and interobserver ICC of 0.755 and 0.831, respectively. The mSASSS scores were more reliable, with ICC of 0.874 and 0.941, respectively. Both scoring systems correlated significantly with BASMI (p = 0.01), while only the mSASSS showed a significant correlation (p = 0.02) with BASFI. With regards to sensitivity to change, it was found that mSASSS classified the highest percentage of patients with more changes than the BASRI-spine (mSASSS: 35.8% vs. BASRI spine: 15.8%). The ES analysis also suggested that the mSASSS was more responsive than BASRI-spine. Concerning feasibility, the BASRI-spine takes less time for scoring. CONCLUSION: We have shown that the mSASSS offers advantages in measurement properties and is the most appropriate method by which to assess progression of structural damage in AS. PMID- 17417994 TI - Adaptation and validation of a telephone questionnaire--Serbian version for case detection of rheumatoid arthritis and spondyloarthropathy (multicentric Eular study). AB - OBJECTIVE: To adapt and validate a telephone questionnaire for case detection of rheumatoid arthritis (RA) and spondyloarthropathies (SpA) in the Serbian population. METHODS: A questionnaire, developed by the French Society of Rheumatology and successfully tested in France, was adapted to the Serbian language using a cross-cultural adaptation process. It was validated in 150 patients: 50 with RA, 50 with SpA and 50 with degenerative rheumatic disorders. They were recruited from Institute of Rheumatology in Belgrade, hospital registry, years 2001 and 2002. The questionnaire validity was assessed in reference to clinical diagnosis and ACR 1987 and ESSG 1991 classification criteria. A logistic regression model was used for RA-control and SpA-control comparison to identify the set of items that best discriminates these groups. RESULTS: Cross-cultural adaptation of the Questionnaire was successfully achieved, verifying its equivalence with the original (semantic, idiomatic, experiential, conceptual). According to the logistic regression, two items selected for RA provided 92.1% agreement when using either clinical diagnosis or ACR classification criteria as a standard. SpA-control comparison included five items providing 96.8% agreement with clinical diagnosis and four items providing 94.1% agreement with ESSG criteria. Results of the present study are similar to those found in the French study. CONCLUSION: Validation results of the telephone questionnaire, translated and adapted to the Serbian language, confirm that it can be used as a detection tool for RA and SpA cases in the population of Serbia, whose diagnoses would have to be further confirmed. PMID- 17417995 TI - Switching to etanercept in patients with rheumatoid arthritis with no response to infliximab. AB - TNF-alpha is thought to play a pivotal role in the initiation and perpetuation of the chronic inflammatory process in rheumatoid arthritis. TNF-alpha blockers such as infliximab and etanercept are currently used in the treatment of active rheumatoid arthritis (RA) when traditional DMARDs have failed and are effective in a significant proportion of patients. However, about one third are non responders to anti-TNF-alpha. The aim of this study was to verify whether rheumatoid patients, after failing infliximab, can benefit from etanercept. We analysed 18 patients with active RA with no response to at least 3 DMARDs and where infliximab therapy had failed. The patients had received infliximab associated with methotrexate: eleven of them did not show any significant response, while seven patients, after a good response, relapsed. Etanercept was then started. EULAR criteria of response were used with calculation of activity index DAS28 at baseline, after 2 weeks, 3 months and every third month until last follow-up. A moderate or good response was achieved with etanercept in 13 out of 18 patients. From our experience, etanercept can be considered as a good alternative choice when infliximab has failed. PMID- 17417996 TI - A study of the arthritis pattern in primary Sjogren's syndrome. AB - OBJECTIVE: To study the frequency and pattern of arthritis in primary Sjogren's syndrome (pSS), and its association with clinical and immunological factors. METHODS: 102 patients with pSS diagnosed according to the preliminary European Classification Criteria were examined yearly for 4.5 years in a prospective study design. Arthralgia and arthritis were registered during the 459 patient-years observation period. RESULTS: Arthralgia was reported by 75 patients (73.5%) and arthritis was demonstrated in 18 patients (17.6%) during the observation period. The most commonly affected joints were ancles (n = 7), MCP joints (n = 6), shoulders (n = 6), MTP joints (n = 6) and wrists (n = 5). Symmetrical bilateral arthritis were most commonly observed in ancles (4 patients) and wrists, shoulders and MTP joints. Five patients had longstanding arthritis observed at more than one clinical examination, and one developed seronegative rheumatoid arthritis. Arthralgia/arthritis was not correlated to any clinical or immunological factors, and usually ESR and CRP were normal when arthritis was observed. CONCLUSION: Arthritis in pSS is usually mild, resolving, and unrelated to other clinical and immunological factors. A typical pattern is uni- and bilateral arthritis in the ankles, but joints in hands, feet and shoulders may also be affected. PMID- 17417997 TI - Fucosylation in synovial fluid as a novel clinical marker for differentiating joint diseases--a preliminary study. AB - OBJECTIVE: To investigate fucosylation of synovial fluid glycoproteins in patients with rheumatoid arthritis (RA), juvenile arthritis (JIA), gonarthrosis (GA) and reactive arthritis (ReA), referred to traumatized knee (TK). METHODS: Synovial fluid glycoproteins were separated by SDS-PAGE and either silver stained or blotted onto nitrocellulose and probed with the fucose-specific Aleuria aurantia lectin. Five bands were chosen for densitometric analysis. Total fucose content and density of fucosylated epitopes were analyzed. RESULTS: Fucose content was elevated in all patient groups and almost all bands, comparing to TK. The density of fucosylated epitopes was increased in the 42-kDa band of RA and JIA cases, and lowered in the 26-kDa band of RA and JIA, but not in GA. In all RA cases FR 42-kDa > FR 26-kDa. The relation was opposite in 8 out of 9 GA cases. CONCLUSION: The density of fucosylated epitopes differs significantly in particular glycoproteins of synovial fluid in joint diseases and may be of potential diagnostic value in differentiating diseases of inflammatory and degenerative origin. PMID- 17417998 TI - Failure to over express MHC-CLASS-1 on muscle biopsy in a case of amyopathic juvenile dermatomyositis. AB - The concept of amyopathic dermatomyositis or dermatomyositis sine myositis, is contentious, particularly within paediatrics. We report an 8-year-old girl presenting with dermatological dermatomyositis without muscle weakness. Muscle biopsy changes are described, in particular, the absence of MHC class 1 over expression. This supports the concept of amyopathic dermatomyositis as a subgroup of juvenile dermatomyositis (JDM) and suggests that immunohistological analysis may be a valuable in excluding a myositic element in such cases. PMID- 17417999 TI - Pulmonary sarcoidosis in a patient with ankylosing spondylitis treated with infliximab. AB - We present a case of a 34 year-old male diagnosed with ankylosing spondylitis. After two years of treatment with infliximab, the patient developed a clinical picture compatible with stage II thoracic sarcoidosis. These findings resulted in the interruption of infliximab therapy. The patient was not administered new treatment since respiratory function testing did not confirm harmful repercussions. After a follow-up of 1 year, the patient is asymptomatic and radiologic tests show complete resolution of pulmonary infiltrates and mediastinal and bilateral hilar lymphad- enopathy. PMID- 17418000 TI - Clustering of cardiovascular risk factors in rheumatoid arthritis: the rationale for using statins. AB - Atherosclerosis may be more prevalent and more extensive in individuals with rheumatoid arthritis (RA) compared with the general population. Despite the fact that traditional and novel cardiovascular disease (CVD) risk factors are clinically important in these patients, it seems that inflammation--a key feature of RA--plays a crucial role in atherogenesis. Reducing the CVD burden in patients with RA is a more complex process than in the general population, mostly due to inadequate inflammation suppression as well as multiple concomitant drug therapy. Furthermore, there is no current consensus on whether RA patients should be treated as individuals at high-risk for vascular events. Statins have proved their efficacy in reducing CVD events in the general population. Despite the fact that they are not specifically indicated in RA, there is evidence supporting a beneficial effect on CVD risk factors as well as disease activity and progression. The present review considers the traditional and novel as well as the RA-specific CVD risk factors. The current evidence supporting the use of statins in this patient population is also discussed. PMID- 17418001 TI - Persistence of protein oxidation products and plasma antioxidants in juvenile idiopathic arthritis. A one-year follow-up study. AB - OBJECTIVE: Plasma protein oxidation products and blood antioxidants, like superoxide dismutase (SOD), glutathione peroxidase (GPx) and total antioxidant status (TAS) were investigated in children with juvenile idiopathic arthritis (JIA) in a year follow-up study. METHODS: Carbonyl group content within plasma proteins, activity of red blood cell SOD and GPx, as well as the blood TAS level were determined in 14 children with JIA twice, namely at the admission to the hospital (Time 0 = T0) and then after a year of treatment (Time 1 = T1). RESULTS: An increased level of plasma protein carbonyls was observed in both assessments (T0 and T1) as compared to control. However there was no significant difference in plasma carbonyls level between the initial (T0) and final (T1) examination of the patients. Similarly, SOD activity was higher in children with JIA as compared to control subjects and did not change significantly after a year of follow-up. Red blood cell GPx activity remained within the normal range throughout the study. Interestingly, the blood TAS level was initially comparable to control and rose significantly after the year of treatment. CONCLUSION: A level of plasma protein oxidation products remains significantly higher in children with JIA as compared to healthy subjects. The lack of accumulation of plasma protein carbonyls may result from efficient proteolysis in childhood and/or adaptive increase of the blood TAS level in the course of effective anti-inflammatory therapy. Analysis of plasma oxidative stress markers and antioxidant potential of the blood might be helpful in monitoring the clinical treatment of children suffering from JIA. PMID- 17418002 TI - Effectiveness and safety of leflunomide in the clinical practice. A different experience. PMID- 17418003 TI - Level of agreement between rheumatologists on US image acquisition using a 3D volumetric probe. PMID- 17418005 TI - Appearance of rheumatoid nodules following anti-tumor necrosis factor alpha treatment with adalimumab for rheumatoid arthritis. PMID- 17418006 TI - Isolated chronic severe neutropenia in eosinophilic fasciitis. Failure of response to granulacyte colony-stimulating factor. PMID- 17418007 TI - Visceral leishmaniasis in a patient with systemic juvenile arthritis treated by IL-1RA agonist (Anakinra). PMID- 17418008 TI - [Efficacy in advanced breast cancer should be evaluated by time to progression]. PMID- 17418009 TI - [BRCA1 1100delAT is a recurrent mutation in Chinese women with familial breast cancer]. AB - OBJECTIVE: Previous investigation on BRCA1 gene mutations in thirty-five breast cancer patients with affected relatives in Shanghai identified four germ-line mutations (1100delAT, IVS17-1G > T, IVS21+1G > C and 5640delA). To our knowledge, up to now, no founder mutation in BRCA1 gene has been identified in Chinese mainland population. The aim of this study was to investigate whether there are recurrent mutations or 'founder mutations' in Chinese mainland population. METHODS: Peripheral blood samples were collected from 60 breast cancer patients with at least one first-degree relative affected with breast cancer from Shanghai, Jinan, Qingdao, and Shenyang. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR) to analyze the 4 germ-line mutations (1100delAT, IVS17-1G > T, IVS21+1G > C and 5640delA) discovered previously: the amplicons were analyzed by denaturing high-performance liquid chromatography (DHPLC), and those with abnormal chromatographic profiles were confirmed by direct sequencing. Four BRCA1-linked markers were used to do allelotype analysis. RESULTS: Only the 1100delAT mutation in BRCA1 gene recurred in two unrelated individuals. Allelotype analysis showed that the two individuals who carried the 1100delAT mutation shared the same allelotype at 4 sites: D17S855, D17S1322, D17S1326, and D17S1327, which was different from the allelotype of the patients who carried the mutation at the site D17S1322 previously reported in Shanghai population. This recurrent mutation gave an overall prevalence of 3.16% (3/95) in all of our investigated population. A novel mutation, 5589del8, was found in one case. CONCLUSION: Recurrent mutation is found in Chinese mainland familial breast cancer patients for the first time. 1100delAT mutation may be a hotspot in BRCA1 gene in Chinese population. Whether this mutation is a founder mutation in the Northern Chinese community need further investigation. PMID- 17418010 TI - [Relationship between LKB1 gene and invasion-related factors of breast cancer cells]. AB - OBJECTIVE: To investigate the relationship between LKB1 gene and invasion of breast cancer cells. METHODS: Human breast cancer cells of the line MDA-MB-435 were cultured and transfected with plasmids with or without LKB1 gene. The clone of the transfected MDA-MB-435 cells with high expression of LKB1 was called MDA MB-435/LKB1 (H), and that with low expression of LKB1 was called MDA-MB-435/LKB1 (L). The MDA-MB-435 cells transfected with blank vector was called MDA-MB 435/vec. MDA-MB-435 cells without transfection were used as controls. RT-PCR was used to detect the mRNA expression of the invasion-related factors of breast cancer: matrix metalloproteinase (MMP)-2, MMP-9, vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF), and Western blotting was used to detect the protein expression of these factors. Gelatin zymography was used to expression of the secretive MMP-2 and MMP-9. Transwell test was used to examine the membrane penetration of the different MDA-MB-435 cells. RESULTS: The invasion rate was (47.6 +/- 1.3)% in the MDA-MB-435 cells, (45.6 +/- 1.2)% in the MDA-MB-435/vec cells, both significantly higher than those of the MDA-MB 435/LKB1 (H) and MDA-MB-435/LKB1 (L) cells [(18.1 +/- 1.0)% and (22.4 +/- 1.8)% respectively, all P < 0.01], with significant difference between the latter 2 groups (P < 0.05). The mRNA expression and protein expression of MMP-2, MMP-9, VEGF, and b-FGF were all significantly lower in the MDA-MB-435/LKB1 cells. Gelatin zymography showed that the secretive MMP-2 and MMP-9 were expressed at the protein level significantly lower in the MDA-MB-435/LKB1 cells. CONCLUSION: LKB1 gene inhibits the invasion of breast cancer cells. PMID- 17418011 TI - [Spleen tyrosine kinase (L) suppresses breast cancer development by downregulating the expression of cyclin D1, ID1, B-myb and Fra1]. AB - OBJECTIVE: To screen the target genes regulated by the transcriptional repressor Syk (L) so as to explore the mechanism of Syk (L) in suppressing breast cancer. METHODS: Adeno-X expression system from Clontech Company was used to construct the adenovirus-Syk. Syk negative breast cancer cells of the line MB231 were cultured and infected with adeno-Syk (L), adeno-Syk (S), and the control adeno Lac Z respectively. cDNA microarray was used to screen the genes regulated by Syk (L) and Syk (S). Northern blotting was used to examine the cDNA microarray results. RESULTS: The transcriptional repressor Syk (L) was able to down-regulate the expression of the oncogenes ID1, cyclin D1, Fra1, and B-myb in the breast cancer cells. Northern blotting confirmed this regulation. CONCLUSION: Syk (L) suppresses the progression of breast cancer by down regulating the expression of the oncogenes ID1, cyclin D1, Fra1, and B-myb. PMID- 17418012 TI - [Application of breast conservative therapy in breast cancer patients with nipple discharge]. AB - OBJECTIVE: To study the feasibility of breast conserving treatment in breast cancer patients with nipple discharge. METHODS: Retrospective analysis were carried out in 7 cases of breast cancer patients with nipple discharge who were treated by central segmental resection with latissimus dorsi myocutaneous transfer. RESULTS: No flap necrosis is happened, and cosmetic outcome was excellent in all 7 patients. Average follow-up was 7 months, there was no local recurrence and distant metastasis. CONCLUSION: Central segmental resection with latissimus dorsi myocutaneous transfer as the local treatment strategy for breast cancer patients with nipple discharge is feasible and safe. PMID- 17418014 TI - [Total mastectomy and axillary dissection with conservation of the nipple-areola complex, and immediate reconstruction with artificial prosthesis in the treatment of early breast cancer]. AB - OBJECTIVE: To study the feasibility of total mastectomy and axillary dissection with conservation of the nipple-areola complex, and immediate reconstruction with artificial prosthesis in the treatment of early breast cancer. METHODS: The 20 patients with breast cancer (TNM stage I, 6 patients; stage II, 14 patients) underwent the total mastectomy axillary dissection with conservation of the nipple-areola complex, and immediate reconstruction with artificial prosthesis. Post-operation they were given chemotherapy, radiotherapy, endocrine therapy. RESULTS: The mean follow-up for the entire group of 20 patients was 52.3 months, eighteen patients were more than 36 months. All patients were still alive, without local or regional recurrences. One patient suffered from multiple metastasis of liver and bone nine months after the operation, who has living five years now. CONCLUSION: Total mastectomy and axillary dissection with conservation of the nipple-areola complex, and immediate reconstruction with artificial prosthesis on selected patients with breast cancer is an oncologically acceptable procedure with superior cosmetic results, without increasing the risk of local recurrences, complications and influencing the effects of postoperative adjuvant treatment. PMID- 17418015 TI - [Association between melanocortin 4 receptor gene polymorphisms and obesity]. AB - OBJECTIVE: To investigate the association between the polymorphisms of melanocortin-4 receptor (MC4R) gene and obesity. METHODS: The genotypes of three polymorphisms, nt-216C/T, nt-178A/C, and Val103Ile, were determined through DNA sequencing in 563 Chinese from Shanghai, including 258 individuals with body mass index (BMI) over 30 kg/m(2) and 305 individuals with BMI less than 23 kg/m(2). Height and body weight were measured to calculate BMI. Blood pressure was measured. Waistline, hipline, and femoral waist to calculate the waist-to-hip ratio and waist-to-femoral ratio were measured. Oral glucose tolerance test was conducted to detect the plasma glucose level during fasting condition and 120 minute after glucose loading. The levels of serum lipids, including total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol. RESULTS: (1) The frequencies of nt-216C/T, nt 178A/C, and Val103Ile were all less than 5%. (2) Logistic regression showed that the Val103Ile variant was an independent risk factor for obesity (OR = 0.414, P = 0.040). The frequency of Ile was less in the obese individuals compared with the controls. (3) In the controls and obese individuals, no association was detected between the genotypes and phenotypes. CONCLUSION: Val103Ile variant of MC4R gene is associated with obesity in Chinese. PMID- 17418016 TI - [Association of C1019T polymorphism in the connexin 37 gene and coronary artery disease in Chinese Han population]. AB - OBJECTIVE: To investigate the association between the connexin 37 (CX37) C1019T polymorphism and the susceptibility to coronary artery disease (CAD) in northern Han population of China. METHODS: A total of 514 CAD patients and 400 healthy controls diagnosed by angiography were genotyped by using polymerase chain reaction-restriction fragment length polymorphism and polyacrylamide gel electrophoresis. RESULTS: The genotype frequencies of CC, TC and TT in the CX37 C1019T polymorphism was 22.37%, 53.31% and 24.32% in CAD patients, 17.75%, 46.50% and 35.75% in the controls respectively (P = 0.0007). The frequency of the CX37 C allele in CAD patients was significantly higher than that of the control group (49.03% vs 41.00%, OR = 1.38, 95% CI = 1.15 - 1.66, P = 0.0006). The frequency of the C allele carriers (CC + TC) was 75.68% in the CAD group and 64.25% in the control group (P = 0.0002). Compared with the TT homozygote, the CAD risk was significantly increased in the carriers of C allele (CC + TC) (OR = 1.73, 95% CI = 1.30 - 2.30). Subsequent stratified analysis revealed that the frequency of C allele was significantly higher in the male CAD patients than in the male controls (49.37% vs 39.60%, OR = 1.49, 95% CI = 1.18 - 1.89, P = 0.0009). The CAD risk was nearly two-fold increased in the carriers of C allele (CC + TC) than in the TT homozygote (95% CI = 1.38 - 2.78). However in the female population, there was no difference in the CAD risk between the carriers of (CC + TC) type and the TT homozygote (P = 0.24). CONCLUSION: The C allele in the CX37 gene might be associated with the susceptibility to CAD and potentially plays an important role in the manifestation of coronary atherosclerosis among Chinese. PMID- 17418017 TI - [Relations of platelet phospholipid fatty acids to thrombotic risk factors in middle-aged and geriatric patients with hyperlipidemia]. AB - OBJECTIVE: To investigate the relations of platelet phospholipid fatty acids to thrombotic risk factors in the middle-aged and geriatric patients with hyperlipidemia in the metropolitan area of Hangzhou, Zhejiang province. METHODS: A questionnaire survey was conducted among 81 patients with hyperlipidemia, 50 males and 31 females, aged (57 +/- 8), and 65 healthy controls, 43 males and 22 females, aged (58 +/- 9) to collect the data about height, weight, and diet. Peripheral venous blood samples were collected to examine the total cholesterol, triglyceride, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), homocysteine (Hcy), 6-keto-prostaglandin F(1alpha) (6-keto-PGF(1alpha)), and thromboxane B(2) (TXB(2)) were examined by standard methods. Serum thrombotic risk factors including homocysteine and Thromboxane B(2) were determined by standard methods. Platelet phospholipid fatty acids were examined by gas chromatography. The correlation between the serum thrombotic risk factors (Hcy, TXB(2), and 6-keto-PG F1a) was analyzed by multivariate linear regression. RESULTS: There were no significant differences in platelet phospholipid fatty acids between the patients with hyperlipidemia and the healthy controls. Serum Hcy was significantly negatively correlated with docosahexaenoic acid (DHA) and the ratio of n-3 PUFA (polyunsaturated fatty acids)/n-6 PUFA (r = -0.277 and -0.231, both P < 0.01). The level of serum TXB(2) was significantly positively correlated with arachidonic acid (r = 0.176, P < 0.05), and significantly negatively correlated with DHA (r = -0.209, P < 0.01), eicosapentaenoic acid (EPA) (r = -0.194, P < 0.05), and n-3 PUFA/n-6 PUFA (r = 0.238, P < 0.01). CONCLUSION: Increasing the ratio of n-3 PUFA/n-6 PUFA in platelet phospholipid may potentially decrease the thrombotic risks such as Hcy and TXB(2) and provide a reference for diet selection. PMID- 17418018 TI - [Value of 18F-FDG metabolic imaging in diagnosis and treatment of head and neck tumors and its mechanism study]. AB - OBJECTIVE: To study the value of (18)F-FDG dual-head tomography with coincidence (DHTC) and single photon emission computerized tomography (SPECT) coincidence imaging in diagnosis and treatment of head and neck tumors and mechanism thereof and analyze the value of glucose transporter proteins in the mechanism of increased uptake glucose of head and neck malignant tumor. METHODS: Twenty-five patients with head and neck tumors were examined by CT or MRI and underwent (18)F FDG DHTC and coincidence imaging. The results of these 2 different methods were compared. Fresh tissues of 38 patients with malignant tumors of the head-and-neck underwent RT-PCR and immunohistochemical examination. RESULTS: The sensitivity, specificity, and accuracy of (18)F-FDG coincidence imaging and registration with integrated CT (SPECT/CT) in diagnosis of the head and neck tumors was 100.0%, 87.5%, and 96.0% respectively, all significantly higher than those of the anatomical imaging (64.7%, 50.0%, and 60.0% respectively, all P < 0.05). For the lesions on the same site, SPECT/CT could diagnose exactly the primary tumor site of neck metastasis in four cases and diagnose the malignancy or benignancy of other four cases that anatomical imaging (CT/MRI) could not diagnose exactly. (18)F-FDG coincidence imaging and registration with integrated CT could find extra lesions of tumors. The results of RT-PCR and immunohistochemistry showed that the mRNA expression and protein expression of Glut-1 and Glut-3 were higher in the head and neck cancer than that in the normal tissue of head and neck or in the adjacent tissue (all P < 0.05). CONCLUSION: (18)F-FDG coincidence imaging and registration with integrated CT can be as a prospective tool that can judge the malignancy or benignancy of head and neck tumor, and stage and classify the tumor, and distinguish recurrence or necrosis of tumor after treatment by surgery or radiotherapy, and detect unknown primary tumor. The abnormal expressions of Glut-1 and Glut-3 may be correlated with the increased uptake of glucose of head and head cancer. PMID- 17418019 TI - [Effects of preconditioning and postconditioning on emergency percutaneous coronary intervention in patients with acute myocardial infarction]. AB - OBJECTIVE: Postconditioning is a novel approach to myocardial protection during ischemia reperfusion. To investigate the effects of preconditioning and postconditioning on coronary blood flow velocity and prognosis of the patients with acute myocardial infarction (AMI) undergoing emergency percutaneous coronary intervention (PCI). METHODS: Ninety-six patients with AMI underwent revascularization by primary PCI within 12 h after the onset. The 35 patients with preinfarction angina were treated with preconditioning (Precond group). The other 61 patients without preinfarction angina were randomized into two groups: 29 patients undergoing PCI without postconditioning [reinfusion (IR) group], and 32 patients undergoing PCI with postconditioning (3 cycles of reinfusion for 30 s/re-occlusion for 30 s beginning within 1 minute after reinfusion, Postcond group). Corrected thrombolysis in myocardial infarction (TIMI) frame count (CTFC) was used to evaluate the velocity of coronary blood after PCI. Creatine phosphokinase (CK), CK-MB, and malondialdehyde (MDA) were measured before and after PCI. Wall motion score index (WMSI) was assessed by two-dimensional echocardiography before and 8 weeks after angioplasty. RESULTS: There were no significant differences between the three groups with regard to age, sex, presence of angiographically visible collaterals, and elapsed time from the onset of symptoms until perfusion. The CTFC values of the patients of the Precond and Postcond groups were both 27 +/- 6, 27 +/- 6, both significantly faster than that of the patients of the IR group (31 +/- 7, both P < 0.05). The CK peak values of the Precond and Postcond groups were 1242 U/L +/- 801 U/L and 1237 U/L +/- 813 U/L respectively, both significantly lower than that of the IR group (1697 U/L +/ 966 U/L, P < 0.05). The CK-MB peak values of the Precond and Postcond groups were 122 U/L +/- 78 U/L and 117 U/L +/- 76 U/L respectively, both significantly lower than that of the IR group (172 U/L +/- 93 U/L, P < 0.05). The MDA of the Precond and Postcond groups at all time points were all significantly lower than that of the IR group (all P < 0.05). The WMSI values 8 weeks after PIC of the Precond and Postcond groups were 1.2 +/- 0.2, and 1.2 +/- 0.2 respectively, both significantly lower than that of the IR group (1.4 +/- 0.3, P < 0.05). CONCLUSION: A simple and operative procedure to improve the coronary blood flow velocity and heart function and reduce the production of free oxygen radicals, postconditioning can be used clinically widely so as to better the prognosis of AMI. PMID- 17418020 TI - [Selective laser trabeculoplasty in the treatment of primary open-angle glaucoma]. AB - OBJECTIVE: To investigate the efficacy and safety of selective laser trabeculoplasty (SLT) in the treatment of primary open angle glaucoma (POAG). METHODS: In a prospective non-randomized clinical study, 63 patients (85 eyes) with POAG and medically uncontrolled intraocular pressure (IOP) underwent SLT. A total of 50 +/- 5 adjacent but nonoverlapping spots were placed over inferior 180 degrees of the trabecular meshwork using a 532 nm, Q-switched, Nd: YAG laser at energy levels ranging from 0.6 - 1.8 mJ per pulse. After SLT, the eyes continued to receive the identical drug regimen. All patients were observed before and 1, 2 hours, 1, 7 and 14 days, and 1, 2, 3, 6, 9, 12, 18, and 24 months after the treatment. RESULTS: The average pre-operative IOP was (25 +/- 4) mm Hg (1 mm Hg = 0.133 kPa). The mean IOP reduction from baseline were 8.1 mm Hg (32.0%) 1 day after the SLT, 5.6 mm Hg (22.1%) 7 days after the SLT, 4.7 mm Hg (18.6%) 14 days after the SLT, 5.5 mm Hg (21.7%) 1 month after the SLT, 5.1 mm Hg (20.2%) 2 months after the SLT, 5.9 mm Hg (23.3%) 3 months after the SLT, 5.2 mm Hg (20.6%) 6 months after the SLT, 4.0 mm Hg (15.8%) 9 months after the SLT, 4.2 mm Hg (16.6%) 12 months after the SLT, 3.8 mm Hg (15.0%) 18 months after the SLT, and 3.3 mm Hg (13.0%) 24 months after the SLT (P < 0.01 or 0.05). Adverse reactions were minimal, including conjunctival injection, mild anterior chamber reaction, and transient pressure spike. CONCLUSION: SLT is a safe, and effective method of reducing IOP in POAG patients. PMID- 17418021 TI - [Surgical treatment of diffused osteoradionecrosis of temporal bone in cases with nasopharyngeal carcinoma after radiotherapy]. AB - OBJECTIVE: To investigate the effective treatment method of osteoradionecrosis (ORN) of temporal bone in the patients with nasopharyngeal carcinoma (NPC) after radiotherapy. METHODS: Eight NPC patients (8 ears) with ORN of temporal bone accepted surgical treatment, 2 ears undergoing radical mastoidectomy, 2 ears undergoing extensive radical mastoidectomy, 5 ears undergoing radical mastoidectomy and obliteration with transferring local vascularized fascia flaps. RESULTS: Five of the 8 ears (62.5%) achieved dry ear, including 4 ears undergoing radical mastoidectomy and obliteration with vascularized fascia flaps, and 1 ear undergoing radical mastoidectomy. Two of the 8 ears (25%) still had infection and were not fully epithelized, but without sequestration, including 1 ear undergoing mastoidectomy and obliteration with vascularized fascia flaps, and 1 ear undergoing extensive radical mastoidectomy. One of the 8 ears (12.5%) which had received radical mastoidectomy needed revision surgery because of re sequestration. CONCLUSION: The surgical treatment for diffused ORN of temporal bone by radical mastoidectomy and obliteration with local vascularized flaps is effective. The main objective of the surgery is get excellent drainage and prevention of complications. PMID- 17418022 TI - [Analysis of etiology of hemorrhagic cystitis after allogeneic hematopoietic stem cell transplantation]. AB - OBJECTIVE: To analyze the etiology and clinical features of late onset hemorrhagic cystitis (LOHC) after allogeneic hematopoietic stem cell transplantation (HSCT). METHODS: The medical records of 200 patients undergoing allogeneic HSCT from 2004 to 2005 were analyzed retrospectively. RESULTS: HC developed in 57 patients within 180 days after the transplantation with a cumulative incidence of 28.8%. The etiology of 31 patients (54.39%) was infection, caused by infection of cytomegalovirus (CMV) or adenovirus and cured by anti-virus therapy, thus the cause of disease could be classified as infection agent. Viremia was seen in 12 patients (21.53%) with CMV and disappeared in urine after anti-virus therapy but bleeding still persisted. For these patients the cause of disease was classified as infection agent combined with non-infection factor. Evidence of infection agent could not be discovered in 14 patients (24.56%) and they failed to respond to anti-infection therapy. For them the cause of disease was classified as non-infection agent. 13 patients with refractory HC underwent tentative treatment with corticosteroids, 9 of them achieved a complete remission, 2 of them achieved partial remission, and 2 of them remained non responsive. CONCLUSION: LOHC after allo-HSCT is a common complication and caused by multiple factors. Differentiation of the possible causes may benefit its clinical outcome. PMID- 17418023 TI - [Study on association of BAFF receptors gene expression and primary biliary cirrhosis]. AB - OBJECTIVE: To explore the relationship between expression level of B cell activating factor belonging to the TNF family (BAFF) receptor (BAFF-R) gene family and primary biliary cirrhosis (PBC). METHODS: With 18S rRNA as control, real-time fluorescent semi-quantification reverse transcriptional PCR was established, according to the difference of threshold cycles (DeltaCt) of target genes and 18S rRNA. B cell maturation antigen (BCMA), transmembrane activator and CAMI-interactor (TACI) and BAFF-R mRNA of peripheral blood mononuclear cells (PBMCs) in 30 healthy people and 30 PBC patients were measured. The correlation between their gene expression levels and the concentrations of IgM, ALP and GGT was analyzed. RESULTS: The relative expression level of BCMA mRNA in patients with PBC was 8.6 folds higher than that of control (P < 0.05). In contrast, PBC patients showed relatively lower TACI mRNA level than control. For BAFF-R mRNA, there was no significant difference between patients and control. The gene expression of BCMA showed a close correlation with IgM, GGT and ALP (all P < 0.01), while BAFF-R showed no significant correlation with them (all P > 0.05). The correlation between TACI and IgM was significant (P < 0.05), but not for GGT or ALP (both P > 0.05). CONCLUSION: The gene expression of PBMCs in PBC patients is significantly elevated for BCMA and decreased for TACI, indicating that the pathogenesis of PBC is closely related to enhanced humoral immunity and broken tolerance. PMID- 17418024 TI - [Quantitative analysis of sensory neurons innervating the muscle levator ani in rats with anorectal malformation]. AB - OBJECTIVE: To quantitatively observe the sensory neurons innervating the levator ani muscle and explore the cause of poor post-operative anorectal function in patients with anorectal malformation (ARM). METHODS: Combining the microsurgery and microinjection techniques, we investigated the deficiency of sensory neurons of the spinal cord in fetal rats with anorectal malformation by injecting the retrograde tracer fluorogold (FG) into the muscle levator ani. Sixty 11 days pregnant female rats were fed with ethylenethiourea (ETU) so as to cause ARM in the fetuses. When the female rats were 20 days pregnant they underwent hysterectomy and the male fetal rats were taken out. Fluorogold (FG), a tracer, was injected into their levator ani muscles. Then the fetal rats were put back into the uteri. Twenty-four hours later hysterectomy was performed for the second time to take out and kill the live fetuses that had undergone FG injection. Their lumbo-sacral spinal cords were taken out, fixed and made into serial longitudinal sections. Digital camera system and fluorescent microscopy were used to observe the FG-labeled sensory neurons. Fifteen normal female rats were used as controls. RESULTS: The FG-labeled sensory neurons innervating levator ani muscle were located mainly in the posterior root ganglia of the 5 th segment of lumber and the 1 st sacral cord. The number of FG-labeled sensory neurons in the normal control fetuses, fetuses without defect of the ETU-fed female rats, fetuses with low type imperforate anus, and fetuses with high type imperforate anus were 11 804 +/- 2362, 10 429 +/- 708, 2886 +/- 705, and 1026 +/- 425 respectively, the latter 2 number being significantly fewer than that in the fetuses without defects (P < 0.05) and that in the normal controls (P < 0.05). CONCLUSION: Defective sensory neurons innervating the levator ani muscle co-exists with the alimentary tract anomaly in rat fetuses with ARM. The defective development of sensory neurons is an important factor likely to contribute to poor post operative anorectal function despite surgical correction of the ARM. PMID- 17418025 TI - [The modulating effect of proapoptotic protein Bax on the resistance of malignant lymphoma cells to tumor necrosis factor related apoptosis inducing ligand-induced apoptosis]. AB - OBJECTIVE: To elucidate the modulating effect of proapoptotic protein Bax on the resistance of malignant lymphoma cells to tumor necrotic factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis and offer evidence for clinic work. METHODS: Human malignant lymphoma cells of the line CRL and normal HRC cells were cultured and treated by 500 g/L TRAIL (Group T), treated by 10 nmol/L PS-341 (Group P), or pre-treated by 10 nmol/L proteasome inhibitor PS-341 for 1 hour and then treated by 500 microg/L TRAIL (Group P + T). Flow cytometry was used to detect the cell apoptosis. Western blotting was used to detect the expression of Bax protein. Caspase-8 activity was tested by fluorophotometer. Immunoprecipitation method was used to examine the conformation change of Bax protein. RESULTS: The apoptosis index 24 hours after treatment of the CRL cells of Group T was 21%, significantly lower than that of the HRC cells of Group T (32%, P < 0.01). The Bax protein expression amount 24 hours after treatment of the HRC cells of Group T was 1.8 times that of the normal cells, and the Bax protein expression amount 24 hours after treatment of the CRL cells of Group T was only 5/17 that of the normal amount. The apoptotic index 6 hours after treatment of the CRL cells of Group P + T was 54%, significantly higher than those of Groups T and P (both P < 0.01). The caspase-8 activity 6 hours after treatment of the CRL cells of Group P + T was 26.5 micromol.L(-1).h(-1).mg(-1) protein, similar to that of the HRC cells of Group P + T (27.2 micromol.L(-1).h( 1).mg(-1) protein), and significantly higher than those of the other cells (all P < 0.01). The Bax protein expression 6 hours after treatment of the Group P HRC and CRL cells were 2.5 times and 1.2 times that of the control cells. The Bax protein expression of the HRC cells of Group P + T was 3.3 times that of the normal controls, and the Bax degradation of the CRL cells of Group P + T was significantly reduced. The combination treatment of P + T significantly increased the expression of activated Bax. CONCLUSION: Bax degradation plays an important role in the resistance of malignant lymphoma to TRAIL-induced apoptosis. Using proteasome inhibitor can inhibit the protein degradation and overcome the drug resistance. PMID- 17418028 TI - [The clinical research of arthroscopic anterior cruciate ligament reconstruction]. PMID- 17418029 TI - [Progress in epilepsy surgery]. PMID- 17418030 TI - [The incidence and variation of tunnel enlargement after anterior cruciate ligament reconstruction]. AB - OBJECTIVE: To investigate the incidence and variation of tunnel enlargement after anterior cruciate ligament (ACL) reconstruction. METHODS: ACL reconstructions using hamstring tendons were performed in 58 patients (58 knees) in the study. MRI scans were taken in a consistent manner at 1, 3, 6, 12 and 24 months after surgery to measure tibial and femoral tunnel expansion. RESULTS: Femoral tunnel enlargement was observed in 9 knees (9/58, 15.5%); Tibial tunnel enlargement was found in 12 knees (12/58, 20.7%). Of those with enlarged bone tunnels, there was no significant difference of tunnel diameters between 1 and 3 months after surgery (P>0.05). Six, 12 and 24 months postoperatively, the average tunnel diameters were larger than those of 1 or 3 months after surgery (P<0.05), however, no significant difference was found in between the tunnel diameters 6, 12 and 24 months postoperatively either (P>0.05). CONCLUSION: Tunnel expansion mainly occurs during 3 to 6 months after surgery, and it remains basically unchanged between 12 and 24 months postoperatively. PMID- 17418031 TI - [Bone-patellar tendon-bone graft in anterior cruciate ligament reconstruction: allograft versus autograft]. AB - OBJECTIVE: To evaluate the clinical result of arthroscopic anterior cruciate ligament (ACL) reconstruction with either allo- or auto- bone-patellar tendon bone (B-PT-B) grafts. METHODS: From February 2002 to January 2006, 142 of 187 cases of ACL ruptures who received ACL reconstruction with B-PT-B grafts were studied retrospectively. There were 93 male and 49 female whose age was from 15 to 57 years (mean 26 years). Patients were divided into 2 groups by graft selection: 38 patients with autograft and 104 with allograft. Clinical results were evaluated according to IKDC, Lysholm, Irgang and Larson scales. Sub-items of scales such as pain, swelling and laxity were specifically evaluated. RESULTS: All of the patients were followed up with an average of 24 months (range from 6 to 43 months). All grafts were radiographically in good position at the time of follow-up. KT-1000 examination of affected knee showed less than 3 mm anterior translation difference compared with contralateral one's. Allograft group: 85 patients got normal IKDC score (81.7%). Lysholm score 82.8 +/- 8.5, Irgang score 79.2 +/- 7.3, Larson score 86.7 +/- 3.1. Autograft group: 29 patients got normal IKDC score (76.3%). Lysholm score 84.6 +/- 9.5, Irgang score 79.5 +/- 7.6, Larson score 88.9 +/- 6.8. No significant statistical difference was found between the 2 groups in the comprehend scale (P>0.05). CONCLUSIONS: Both autograft and allograft group achieve good results, and the allograft B-PT-B could provide the similar clinical result as autograft, the preliminary result of allograft reconstruction might indicate predictable result in the future. PMID- 17418032 TI - [Factors leading to failure of anterior cruciate ligament reconstruction]. AB - OBJECTIVE: To analyze the factors contributing to the failure of primary anterior cruciate ligament (ACL) reconstruction. METHODS: From November 2001 to July 2006, 13 patients underwent ACL reconstruction revision because of pathological instability in daily activities after primary ACL reconstruction, and the data of the patients were retrospectively analyzed. RESULTS: In this group, 7 cases were reconstructed with bone-patellar tendon-bone (B-PT-B) autografts, 4 cases with hamstring tendon autograft and 2 cases with hamstring tendon allograft. There were malpositioned bone tunnels in 8 cases. Among them 4 cases had a femoral tunnel in the front of the predicted one, and in the other 4 cases, both the femoral and tibial tunnels were in the front of the predicted ones. In 2 cases reconstructed with allograft, one had to receive a twice operation to take out the allograft because of serious rejection, and the other claimed a failure because of the obviously enlarged bone tunnel. In the 2 cases reconstructed with B-PT-B autograft, malposition of the interference screw using to fasten the bone block had been founded in the upper bone tunnel. One case suffered from postoperative infection and had been cured by the anti-biotic treatment after arthroscopic debridement. Then the reconstructed ACL in 7 cases had absolutely ruptured and been absorbed. Four cases had obviously loosen but still partly linked reconstructed ligament. Two cases had a malposited interference screw, and both of them had no fastening function. One case received the lytic operation due to knee stiffness. Two cases received primary bone transplantation and needed to receive a secondary revision operation owing to severe incompletion of spongy bone. The other 10 cases received primary revision. Among them 4 cases reconstructed with STG obtained from the same lateral, 4 cases reconstructed with STG obtained from the opposite lateral, 1 case reconstructed with B-PT-B obtained from the same lateral and 1 case reconstructed with iliotibiali tractus. CONCLUSIONS: There are many reasons leading to failure of ACL reconstruction. Nevertheless, malposition of the bone tunnel, invalid fixation, rejection to allograft and other complications such as the enlargement of the bone tunnel, postoperative infection and adhesion are the major factors that predispose the primary construction to failure. PMID- 17418033 TI - [Arthroscopic anterior cruciate ligament reconstruction guided by fluoroscopy based navigation system]. AB - OBJECTIVE: To introduce the process and outline of fluoroscopy-based navigation system assisted anterior cruciate ligament (ACL) reconstruction, and evaluate its feasibility and accuracy. METHODS: From September 2005 to February 2006, there were 30 cases ACL rupture patients who received fluoroscopy-based navigation system assisted arthroscopy operations for ACL reconstruction (navigation group). At the same time, there were 40 patients who underwent traditional ACL operation (traditional group). For the navigation group, the proper placement of femoral and tibial tunnels was planned preoperatively in standard AP and lateral X-ray view. Intraoperative fluoroscopic images were taken and input into navigation computer system to form the virtual interactive working fields. After placement and registration, signals from patient trackers, which fixed on the distal femur and tibia respectively, and tool trackers, which attached with ACL tibial and femoral guide, were captured by the optic navigation camera and the navigation computer system could pursue the real-time position of the ACL tools and projected into working field to help precise placement of femoral and tibial tunnels. Then results of two groups were observed and evaluated. RESULTS: For navigation group, the mean time extension was 20 min. The tibial tunnel position was measured in all these cases. The tibial tunnel position of navigation group was 45.90% (SD 2.36%), and the traditional group was 41.05% (SD 6.01%). The difference was statistically significant (P < 0.05). CONCLUSION: Fluoroscopy based navigation system assisted ACL reconstruction improves the accuracy and reproducibility of the tunnel placement. PMID- 17418037 TI - [Surgical treatment of lesional temporal lobe epilepsy]. AB - OBJECTIVE: To discuss the presurgical evaluation and surgical treatment of lesional temporal lobe epilepsy (LTLE). METHODS: We retrospectively studied the patients whose MRI or CT showed lesions on one of the temporal lobes among patients who underwent epilepsy surgeries in our institute. All patients were divided into satisfactory and unsatisfactory group according to outcomes after operation. The characteristics of the lesions, neurophysiological features and results of pathologies were analyzed statistically. RESULT: Favorable surgical outcome was obtained in 24 patients, the satisfactory rate was 75%. The lesions of 17 patients were cortical malformations and the satisfactory rate of this group was 65%, which was less favorable than that of tumor group, (87%). 21 patients with their lesions located within the border of standard temporal lobectomy, had better surgical outcome than the others whose lesions were beyond the border (P<0.05). The satisfactory rate of 8 patients with lesions located within mesial structure of temporal lobe was no different compared with that of the others who had lesions outside the mesial structure (P>0.05). There were 19 patients who had consistency of the location of the lesion on MRI with the focal interictal epileptiform discharges on scalp EEG in satisfactory group, while there were only 3 patients in unsatisfactory group (P<0.05). CONCLUSION: For a LTLE patient, epilepsy surgery should be the first choice to be considered. Careful presurgical studies of the lesion, including its location, pathological property and neurophysiological characteristics, were very helpful for improving the surgical outcome. PMID- 17418038 TI - [Clinical study on the therapeutic effects and mechanism of progesterone in the treatment for acute severe head injury]. AB - OBJECTIVE: To evaluate the therapeutic effects of Progesterone (PG) on the patients with acute severe traumatic brain injury, and investigate it's neuroprotective mechanisms. METHODS: Fifth-six patients with acute severe traumatic head injury were divided randomly into two groups: 26 cases were treated with PG and 30 cases were control. Neurological outcome of the patients were assessed using Glasgow Coma Scale (GCS), Glasgow Outcome Scale (GOS), verbal and motor functions scale and Karnofsky Performance Scale (KPS). The serum concentrations of PG, TNF-alpha and 15-F(2t)-isoprostane were measured at day 1, 5 and 10 after trauma. RESULTS: In the two groups, There were no significant difference in the mortality, GCS of acute healing phase, GOS and verbal and motor functions at 10th days after treatment (P>0.05); After follow-up for 3 months, GOS, verbal functions and KPSin the PG treatment group were better than those in the control group (P<0.05); In addition, there was no difference of motor functions in the two groups (P>0.05). At 5th day after trauma, serum 15-F(2t) isoprostane and TNF-alpha levels increased in the control group, but decreased at 10th day after trauma. Compared with the control group serum PG levels increased, serum 15-F(2t)-isoprostane and TNF-alpha levels reduced significantly in the PG treatment group at 5th and 10th day after injury (P<0.05). CONCLUSION: It indicated that successive early application of PG will benefit the patients with acute severe head injury by improving the recovery and reducing the disability, which may be related to its alleviating inflammatory and lipid peroxidation response. PMID- 17418039 TI - [Clinical research about brain oxygen metabolism and neuroelectrophysiology during mild hypothermia in patients with severe head injury]. AB - OBJECTIVE: To observe the changes of brain oxygen metabolism and neuroelectrophysiology after severe brain injury, and the effects of hypothermia on severe brain injury. METHODS: 148 patients with severe brain injury (GCS 3 - 8, admitted within 10 hours from injury) were selected for this study. Patients were divided into 3 groups, Group GCS 7 - 8, Group GCS 5 - 6 and Group GCS 3 - 4. Every group were also randomly assigned to normothermia and hypothermia subgroup. Patients in the hypothermia group were cooled to 32 approximately 34 degrees C. SLSEP, BAEP, P(br)O(2) and rSaO(2) were recorded in each group at the same time. RESULTS: In the Group GCS 7 - 8, N20 in SLSEP, I/V in BAEP and rSaO(2) were improved significantly after mild hypothermia treatment, and P(br)O(2) was decreased by hypothermia; In the Group GCS 5 - 6, N20 in SLSEP, I/V in BAEP and rSaO(2) were improved by hypothermia, and P(br)O(2) was decreased in hypothermia subgroup; In the Group GCS 3 - 4, no significant difference was found. CONCLUSION: Mild hypothermia has a significant effect on patients of GCS 7 - 8 and a doubt effect on patients of GCS 5 - 6. It seem no effect on patients of GCS 3 - 4. Brain oxygen metabolism and neuroelectrophysiology are important to value the therapeutic effect on severe brain injury. PMID- 17418040 TI - [Clinical pathological study on nodal micrometastases of non-small-cell lung cancer]. AB - OBJECTIVE: To investigate the practicability of detecting the micrometastases in lymph nodes of no-small-cell lung cancer (NSCLC) by means of the immunohistochemical (IHC) staining. METHODS: The lymph node samples were taken from the patients with NSCLC during the operations. Firstly, each resulting tissue block was processed for routine paraffin embedding. Then the 6 approximately 10 serial sections were chosen, each 5 microm thick, from every paraffin block of the lymph node. Finally, the first and the second last sections of each lymph node were stained by hematoxylin eosin (HE), and the other serial sections were used for the IHC staining examination with the monoclonal antibody against cytokeratin 19. RESULTS: The paraffin embedded sections of 195 regional lymph nodes from 25 patients with NSCLC were examined by HE staining. Thirty lymph nodes in 9 patients revealed gross nodal metastases, and none of lymph node in 25 patients showed micrometastatic tumor cells. Frozen tissue sections from 135 regional lymph nodes that were staged as free of metastases by HE staining were screened by IHC staining. Thirty-one lymph nodes in 9 patients showed micrometastatic tumor cells. Five of sixteen patients staged as PN(0) had hilum lymph nodal micrometastases, versus four of nine patients with stage PN(1) had mediastinal lymph nodal micrometastases. There was a significant difference between two groups (chi(2)=52.900, P=0.0193). CONCLUSIONS: Conventional HE staining can accurately detect gross nodal metastases in the lymph nodes of patients with NSCLC, but is unfit for detecting lymph nodal micrometastases. IHC staining analysis can significantly facilitate the detection of occult micrometastatic tumor cells in lymph nodes of NSCLC, and its assessment of nodal micrometastases can provide a refinement of TNM stage for partial patients with stage I to II NSCLC. PMID- 17418041 TI - [Observation on the long-term complications after esophageal replacement with colon]. AB - OBJECTIVE: To study the etiology and preventive measures of the long-term postoperative complication after esophageal replacement with colon for esophageal benign disease. METHODS: To review the clinical data of 577 patients with esophageal replacement with colon our department, including 123 cases of esophageal benign disease. Of all, there were 25 cases-time for 11 cases following with severe complication: redundancy and dilated colon 12 cases-time, severe stricture of stoma 4, macrocyst esophagus 2, colon-stomach stoma expansion 4, mechanical obstruction of colon 3. The etiology included iatrogenic and functionality. The therapy included stricture form or resection, redundancy segment resection, obstructed segment solution and stoma resection and form. RESULTS: Eight cases underwent once operation, 2 case twice, 1 case three times. After operation, 9 cases took food normally, 2 improved symptoms obviously. CONCLUSIONS: The iatrogenic and functionality factor contributed to severe complication after esophageal replacement with colon for esophageal benign disease. The preventive measure is followed during operation: cervical esophageal colon anastomosis exceed 2.5 centimeter, abdominal colon-stomach anastomosis reflux, channel width of colon passage, intestinal canal lay up straight. Re operation is best choice to for local stricture, colon expansion, redundancy and dilated colon. PMID- 17418042 TI - [The surgical treatment and prognosis of bilateral sporadic renal tumors]. AB - OBJECTIVE: To evaluate the surgical management and prognostic characteristics of bilateral sporadic renal tumors (BSRTs). METHODS: Twenty-three cases of BSRTs were identified and retrospectively reviewed. The presenting clinical findings, surgical treatment, and prognosis were evaluated. RESULTS: Twelve patients had synchronous tumors and 11 had asynchronous tumors. BSRTs comprised 1.8% of all RCC cases with a male-to-female ratio of 6.67:1 in the same term. The mean age of patients was 56 years. Seventeen patients were preformed bilateral operation (10 had synchronous and 7 had asynchronous tumors), 4 patients were preformed unilateral operation (1 had synchronous and 3 had asynchronous tumors), and 2 patients were not performed operation (asynchronous tumors). Twenty-one patients were followed up (11 had synchronous tumors and 10 had asynchronous tumors, mean 27 months). One and 5-years cancer specific survival rate of BSRTs were 84.6% and 33.5% respectively. The rate of renal failure of synchronous BSRTs (18%) was lower than that of asynchronous BSRTs (70%) after operation (P=0.030). CONCLUSIONS: Prognosis of BSRTs is poor. The key in the treatment of BSRTs is to detect and resect tumor earlier. Our results indicate that nephron sparing surgery should be performed in order to obtain optimal functional preservation of the renal remnant with complete local excision of tumor. PMID- 17418043 TI - [The clinical characteristic of adrenal metastatic tumor]. AB - OBJECTIVE: To analyze the clinical features of adrenal metastasis. METHODS: From January 1993 to December 2004, 103 cases of adrenal metastasis were reviewed. RESULTS: Lung and hepatocellular carcinoma were the most common primary tumor of adrenal metastatic tumor, which about 36.9% (38/103) and 42.7% (44/103) of all cases, followed by renal carcinoma 6.8% (7/103), colorectal carcinoma 4.9% (5/103), stomach carcinoma 3.9% (4/103), breast cancer 1.9% (2/103), unknown primary tumor 2.9% (3/103). Most of these were low differentiation. The mean diameter of adrenal metastasis was 3.9 cm. The mean interval from detection of primary tumor to adrenal metastasis was 9.5 months. And 79.6% (82/103) were detected as a part of multiorgan metastasis. Only 5 cases (4.9%) were presented with pain in the back. There was little characterization of ultrasonography, CT and MRI, color-Doppler and selective arterial imaging showed little blood supply. All of patients were treated with synthetic methods, 16 cases (15.5%) who had undergone adrenalectomy for metastasis disease had a improved survival compared with those non-adrenalectomy. CONCLUSIONS: There is no particular presentation of clinic and imaging, diagnosis depending on history, follow-up and the pathological presentation of primary tumor. There are no standard treatment guidelines for this group of patients. When the primary tumor could be resected or be well controlled, and there is no other evidence of metastasis, adrenalectomy is recommended. Transarterial chemoembolization (TACE) could not actually be performed. PMID- 17418054 TI - Nutritional immunomodulation of acute pancreatitis. AB - Despite the great advances in our understanding of the pathophysiology of acute pancreatitis, no specific therapy has emerged, and treatment remains supportive. In patients with the severe form of the disease, in which mortality remains high at 20% to 30%, the function of the upper gastrointestinal tract is disturbed due to extrinsic compression by the inflamed and swollen pancreas, and normal eating is impossible. Such patients often develop multiple organ failure, necessitating intensive-care management and artificial ventilation for weeks on end. In this setting, protein catabolism will rapidly result in protein deficiency and further complications unless nutritional support is commenced. Recent studies have shown that, despite the risk of disease exacerbation through pancreatic stimulation, enteral feeding is more effective than parenteral feeding in improving outcome. Experimental studies suggest that this can be attributed to its content of specific immunomodulating nutrients, such as glutamine, arginine, and n-3 fatty acids, and by its stabilizing effect on the gut flora through the provision of prebiotics. Further studies are indicated to examine whether dietary enrichment with these substrates, along with regulation of the gut bacteria with probiotics, can improve outcome further. PMID- 17418055 TI - Predicting severity of acute pancreatitis. AB - Severity stratification is a critical issue in acute pancreatitis that strongly influences diagnostic and therapeutic decision making. According to the widely used Atlanta classification, "severe" disease comprises various local and systemic complications that are associated with an increased risk of mortality. However, results from recent clinical studies indicate that these complications vary in their effect on outcome, and many are not necessarily life threatening on their own. Therefore, "severe," as defined by Atlanta, must be distinguished from "prognostic," aiming at nonsurvival. In the first week after disease onset, pancreatitis-related organ failure is the preferred variable for predicting severity and prognosis because it outweighs morphologic complications. Contrast enhanced CT and MRI allow for accurate stratification of local severity beyond the first week after symptom onset. Among the biochemical markers, C-reactive protein is still the parameter of choice to assess attack severity, although prognostic estimation is not possible. Other markers, including pancreatic protease activation peptides, interleukins-6 and -8, and polymorphonuclear elastase are useful early indicators of severity. Procalcitonin is one of the most promising single markers for assessment of major complications and prognosis throughout the disease course. PMID- 17418056 TI - Pancreatic enzyme therapy for pancreatic exocrine insufficiency. AB - Pancreatic exocrine insufficiency with steatorrhea is a major consequence of pancreatic diseases (eg, chronic pancreatitis, cystic fibrosis, severe acute necrotizing pancreatitis, pancreatic cancer), extrapancreatic diseases such as celiac disease and Crohn's disease, and gastrointestinal and pancreatic surgical resection. Recognition of this entity is highly relevant to avoid malnutrition related morbidity and mortality. Therapy for pancreatic exocrine insufficiency is based on the oral administration of pancreatic enzymes aiming at providing the duodenal lumen with sufficient active lipase at the time of gastric emptying of nutrients. Administration of enzymes in the form of enteric-coated minimicrospheres avoids acid-mediated lipase inactivation and ensures gastric emptying of enzymes in parallel with nutrients. Nevertheless, such factors as acidic intestinal pH and bacterial overgrowth may prevent normalization of fat digestion even in compliant patients. The present article critically reviews current therapeutic approaches to pancreatic exocrine insufficiency. PMID- 17418057 TI - Endoscopic retrograde cholangiopancreatography and endoscopic ultrasound for diagnosis of chronic pancreatitis. AB - The diagnosis of chronic pancreatitis is based on altered pancreatic morphology and function. A spectrum of disease exists, and milder forms of disease may be missed by CT but demonstrated by endoscopic retrograde cholangiopancreatography (ERCP) or endoscopic ultrasound (EUS). The accuracy of ERCP and EUS for diagnosis of "minimal change" or "early" chronic pancreatitis is controversial, particularly when the results from these imaging procedures are discordant with each other or with tests of pancreatic function; in some cases ERCP and EUS should be considered indeterminate for diagnosis. This review discusses recent data concerning the accuracy of ERCP and EUS for diagnosis of chronic pancreatitis, the use of EUS fine-needle aspiration for differential diagnosis of pancreatic masses, and the use of EUS and EUS-guided TruCut biopsy for diagnosis of autoimmune pancreatitis. PMID- 17418058 TI - Approach to pancreatic cystic lesions. AB - Pancreatic cystic lesions are detected more frequently as a result of increasing use of high-resolution cross-sectional imaging. The ability of imaging and cyst fluid analysis to predict the biologic behavior of the lesions has aroused much interest recently. A precise diagnosis is critical to help guide further management. The diagnostic criteria and the management principles have evolved dramatically over the past 10 years with the advent of endoscopic ultrasound and fine-needle aspiration. In this review, we summarize the recent publications that have examined the key issues in the diagnosis and management of cystic lesions of the pancreas. PMID- 17418059 TI - Pancreatoscopy: present and future role. AB - Current imaging modalities do not allow for direct access to the pancreatic duct. Because of this limitation, diagnosis and assessment of pancreatic diseases require radiographic imaging and cytologic analysis. Although many of these techniques have excellent specificity for the detection of pancreatic cancer, they offer no therapeutic capabilities and are relatively poor in early detection of cancer and in differentiating chronic pancreatitis from malignancy, particularly when the conditions coexist. Currently available cholangioscopes are too large to access the pancreatic duct without sphincterotomy or balloon dilation, increasing the risk of instrumentation trauma to the pancreas. Novel, dedicated pancreatoscopes have recently been developed and are under investigation in clinical trials. Preliminary evidence suggests that these new technologies allow for direct visualization of the pancreatic ducts, with the potential for forceps biopsy in suspected malignancy and endotherapy for pancreatic calculi. Future and ongoing clinical trials will better define the utility and roles of these new endoscopy tools. PMID- 17418060 TI - The role of endoscopy in noniatrogenic injuries of the liver. AB - Traumatic noniatrogenic biliary injuries, unlike iatrogenic injuries, are usually complex in nature and are frequently associated with other multiorgan trauma and infection. Bile leaks following these injuries are an important source of short- and long-term morbidity. Repeat surgery for primary repair of complex bile leaks is difficult and can be complicated by anastomotic leakage and biliary stricture formation. Endoscopic retrograde cholangiopancreatography (ERCP) was initially used only as a diagnostic technique to guide surgical repair in this setting. However, with the high success rates observed in treatment of iatrogenic bile leaks, ERCP has emerged as a nonoperative treatment option for noniatrogenic biliary leaks as well. Recent data show that ERCP is effective in managing bile leaks after blunt and sharp liver injuries, using transpapillary stenting, endoscopic sphincterotomy, or both, with greater than 80% healing rates. The evidence is not clear regarding which ERCP maneuver - endoscopic sphincterotomy, transpapillary stenting, or both - should be used. PMID- 17418061 TI - Caroli's disease: identification and treatment strategy. AB - Caroli's disease is a rare congenital disease of the liver characterized by cystic dilation of the intrahepatic bile duct. Classic Caroli's disease involves malformations of the biliary tract alone, whereas Caroli's syndrome refers to the presence of associated congenital hepatic fibrosis. Caroli's disease usually presents during childhood and early adulthood. The clinical features of Caroli's disease include jaundice, right upper abdominal pain, and fever due to the associated complications of hepatolithiasis or bacterial cholangitis. Endoscopic or percutaneous cholangiography is the traditional method of diagnosis, but magnetic resonance cholangiopancreatography is emerging as the diagnostic modality of choice. The treatment for Caroli's disease includes supportive care with antibiotics for cholangitis and ursodeoxycholic acid for hepatolithiasis. Surgical resection has been used successfully in patients with monolobar disease. For patients with diffuse involvement, the treatment of choice is orthotopic liver transplantation. PMID- 17418062 TI - Parasitic infestations of the biliary tract. AB - Parasitic infestations of the biliary tract are a common cause of biliary obstruction in tropical countries and can lead to such serious complications as cholangitis and cholangiocarcinoma. Endoscopic therapy has helped in the management of biliary complications caused by these parasites. Ascaris lumbricoides organisms, which normally reside in the jejunum, are actively motile and can invade the papilla, thus migrating into the bile duct and causing biliary obstruction. Endoscopic retrograde cholangiopancreatography is a useful diagnostic tool with potential for therapeutic management of biliary ascariasis. Infestation with Clonorchis sinensis organisms can cause such complications as intrahepatic stones, recurrent pyogenic cholangitis, cirrhosis, cholelithiasis, pancreatitis, and cholangiocarcinoma. Opisthorchis viverrini, Opisthorchis felineus, and Dicrocoelium dendriticum are closely related to C. sinensis and can also cause serious biliary complications. Fascioliasis, caused by Fasciola hepatica and F. gigantica, is a zoonotic helminthiasis that can present as acute hepatic or chronic biliary tract infection. CT, MRI, and ultrasound guidance are useful imaging tools for identifying these parasites and their complications. PMID- 17418063 TI - Neurohormonal regulation of the sphincter of Oddi. AB - The control of sphincter of Oddi (SO) motor activity is complex and involves interactions between the SO smooth muscle with nerves, bioactive agents, and presumably interstitial cells of Cajal. Disturbances in SO motility are known to be related to painful clinical conditions, such as SO dysfunction and acute pancreatitis. Understanding normal SO motility and comparing this to disturbed SO motility patterns may identify mechanisms that could be targeted for future pharmacologic intervention. The effect on SO motility of recently identified neurotransmitters/neuropeptides, such as purines and orexins, is currently being determined. Furthermore, because the control of SO motility is complex, investigations with known bioactive agents, such as cholecystokinin and nitric oxide, are continuing. This review summarizes research investigating SO motility and function performed in 2005 and 2006. PMID- 17418064 TI - Solid state biliary manometry catheter: impact on diagnosis and post-study pancreatitis. AB - Perfusion manometry of the sphincter of Oddi has been the standard for the investigation of patients with presumed sphincter of Oddi dysfunction (SOD). Microtransducer manometry (MTM) of the sphincter of Oddi represents an alternative to perfusion manometry. The technical success and reproducibility of MTM are as good as for perfusion manometry. Current data suggest that the upper limit for normal of basal sphincter of Oddi pressures measured with MTM lies at approximately 35 mm Hg. Pancreatitis risk after MTM in patients with SOD compares favorably with that after perfusion manometry. Low cost and ease of handling make MTM of the sphincter of Oddi an attractive alternative. PMID- 17418065 TI - Daniel Danielopolu--Romanian physiologist. PMID- 17418066 TI - Intrapleural fibrinolytics for pleural infection: partial answers in the discussion of surgical vs nonsurgical treatment. PMID- 17418067 TI - Intrapleural fibrinolytics combined with image-guided chest tube drainage for pleural infection. AB - OBJECTIVE: To present our method of treating pleural infection by using a combination of image-guided chest tube drainage and intrapleural fibrinolytics. PATIENTS AND METHODS: We retrospectively reviewed the medical charts and radiographs of 30 consecutive patients with pleural infection who were seen at our institution from December 15, 1995, to July 1, 2006, 27 of whom received intrapleural urokinase or tissue-type plasminogen activator. End points were death, length of stay in the hospital, and percentage of patients who needed surgery. RESULTS: Placement of chest tubes required image guidance 45.7% of the time. Three patients (10%; 95% confidence interval, 2.1%-26.5%) died of complications from pleural infection. None of the 30 patients (0%; 95% confidence interval, 0%-9.5%) required surgery for treatment of pleural infection. The median hospital length of stay was 11 days. CONCLUSIONS: In the treatment of pleural infection, intrapleural urokinase or tissue-type plasminogen activator in combination with careful image-guided placement of chest tubes is highly effective in resolving the effusion and curing the infection. PMID- 17418068 TI - Asthma treatment in a population-based cohort: putting step-up and step-down treatment changes in context. AB - OBJECTIVE: To assess the frequency and types of visits related to modifications in the intensity of asthma medications. PATIENTS AND METHODS: We retrospectively reviewed the medical records of adults (aged 18-40 years) and children (aged 6-17 years) living in Olmsted County, Minnesota, to evaluate changes in asthma medications by dose and drug class and site and type of visit (routine vs unscheduled) at the time of changes. All records from all visits were reviewed for each patient to identify asthma-related visits at all sites of care from January 1, 2002, through December 31, 2003. RESULTS: The study consisted of 397 adults and children. In 255 patients, 597 asthma medication changes occurred. Step-up changes usually occurred because of an exacerbation or loss of control of asthma and adhered to the medication hierarchy in the national asthma guidelines. Twenty step-up changes involved skipping inhaled corticosteroid (ICS) monotherapy and moving directly to combined ICSs plus a long-acting beta-agonist (LABA). Lack of documentation of asthma symptom frequency or interference with activities made it impossible to determine whether these 'skips' were appropriate. Only 78 physician-directed step-down changes were documented, usually to a lower dose of combined ICSs and LABAs or a move from combined ICSs and LABAs to anti inflammatory monotherapy. Patients initiated additional step-down changes between encounters. Step-down changes occurred at routine or follow-up asthma visits, but the limited number of such visits provided few opportunities for step-down care. CONCLUSION: The continuing episodic-style treatment of asthma aimed at exacerbation management facilitates step-up changes in asthma therapy. The dearth of asthma evaluation visits limited opportunities to step down use of asthma medications and to provide long-term asthma management. PMID- 17418069 TI - Stratification of patient risk based on prostate-specific antigen doubling time after radical retropubic prostatectomy. AB - OBJECTIVE: To assess the risk of local recurrence, systemic progression, and death from cancer among patients who experience biochemical relapse after radical retropubic prostatectomy and to stratify those patients by prostate-specific antigen (PSA) doubling time (DT). PATIENTS AND METHODS: We identified patients who experienced biochemical recurrence (defined as a PSA level < or =0.4 ng/mL) after radical prostatectomy from January 1, 1990, to December 31, 1999, for prostate adenocarcinoma. The PSA-DT was calculated by log linear regression using all PSA values within 2 years of biochemical recurrence. Local recurrence- and systemic progression- free survival and cancer-specific survival were estimated using the Kaplan-Meier method and analyzed by the log-rank test and Cox models. RESULTS: Biochemical recurrence was noted in 1521 (27%) of 5533 men during the follow-up period. Of the 1064 patients with a calculable PSA-DT, 322 (30%) had a PSA-DT of less than 1 year, 357 (34%) had a PSA-DT of 1 to 9.9 years, and 385 (36%) had a PSA-DT of 10 years or more. Patients with a PSA-DT of 10 years or more were less likely to have a higher preoperative PSA level, Gleason score, advanced pathologic stage, and seminal vesicle invasion. Patients with a PSA-DT of 10 years or more were at low risk of local recurrence (hazard ratio [HR], 0.09; 95% confidence interval [CI], 0.06-0.14; compared with patients with a PSA DT of <1 year), systemic progression (HR, 0.05; 95% CI, 0.02-0.13), or death from cancer (HR, 0.15; 95% CI, 0.05-0.43). CONCLUSIONS: Prostate-specific antigen DT is an independent predictor of clinical disease recurrence and mortality after surgical biochemical failure. Risk stratification into high-, intermediate-, and low-risk categories based on the PSA-DT provides helpful clinical information and assists in the development of salvage therapy trials. PMID- 17418070 TI - Monoclonal gammopathy of undetermined significance: predictors of malignant transformation and recognition of an evolving type characterized by a progressive increase in M protein size. AB - OBJECTIVE: To investigate the predictors of monoclonal gammopathy of undetermined significance (MGUS) by considering not only the initial features but also the pattern of evolution of the M protein during the first years after diagnosis. PATIENTS AND METHODS: This study consisted of 359 patients diagnosed as having MGUS at a single institution. Patients who showed a definite and progressive increase in their M protein size according to serum electrophoresis during the first 3 years of follow-up were considered to have evolving MGUS, whereas all others were considered to have nonevolving MGUS. RESULTS: Of the 359 patients, 330 had nonevolving MGUS, whereas 29 fulfilled the criteria for evolving MGUS. Overall, 32 patients developed malignant transformation. The progression rates at 10 and 20 years of follow-up for the evolving and the nonevolving types were 55% vs 10% and 80% vs 13%, respectively. Multivariate analysis revealed that the features significantly associated with a higher risk of progression were evolving MGUS (relative risk [RR], 12.14; P<.001), IgA MGUS (RR, 2.93; P=.006), and M protein concentration (RR, 2.18; P=.04). CONCLUSION: The evolutionary pattern of serum M protein (progressive increasing vs stable) during the first years of follow-up is the most important risk factor for disease progression in patients with MGUS. PMID- 17418071 TI - Further validation of the FOUR score coma scale by intensive care nurses. AB - OBJECTIVE: The FOUR (Full Outline of UnResponsiveness) score is a new coma scale that consists of 4 components (eye, motor, brainstem, and respiration). The scale was recently validated, but variability among nursing staff has been documented. PATIENTS AND METHODS: We prospectively studied the FOUR score in 80 patients with acute neurologic disease in an intensive care unit (ICU) and compared it with the Glasgow Coma Scale (GCS) using 20 experienced and inexperienced neuroscience ICU nurses and nonneuroscience ICU nurses. Each nurse was trained with the use of video examples and instruction cards. Each patient was rated by 2 nurses, with the order randomly assigned. RESULTS: The rater agreement was good to excellent with the FOUR score (weighted kappa: eye, 0.84; respiration, 0.92; brainstem, 0.89; and motor, 0.73) and similar to that for the GCS (weighted kappa: eye, 0.85; verbal, 0.89; and motor, 0.74). Greater average experience in years was associated with less disagreement, but the difference was not statistically significant. CONCLUSION: The FOUR score provides more neurologic information than the GCS. The FOUR score can be used by any ICU nurses, even those with minimal experience. PMID- 17418072 TI - Dysphagia in inflammatory myopathy: clinical characteristics, treatment strategies, and outcome in 62 patients. AB - OBJECTIVE: To assess the clinical characteristics, treatment, and outcome of patients with inflammatory myopathy-associated dysphagia. PATIENTS AND METHODS: We retrospectively reviewed the medical records of all patients with inflammatory myopathy-associated dysphagia seen at the Mayo Clinic in Rochester, Minn, between January 1, 1997, and December 31, 2001. RESULTS: A total of 783 patients were diagnosed as having inflammatory myopathy during the 5-year study period. Of these, 62 patients (41 women and 21 men; mean age, 68.6 years) had inflammatory myopathy-associated dysphagia: 26 with inclusion body myositis (IBM), 18 with dermatomyositis, 9 with polymyositis, and 9 with overlap syndrome. Dysphagia was a presenting symptom in 13 patients (21%), with the highest incidence in the IBM group. Videofluoroscopic examinations revealed pharyngeal pooling and impaired oropharyngeal and cricopharyngeal function. The benefits of swallowing compensation techniques and exercises were difficult to establish. Interventional procedures were performed in 24 patients (39%) and most frequently (62%) in patients with IBM, with cricopharyngeal myotomy being most beneficial. Patients with IBM had the least symptomatic improvement. Overall, 11 patients died during the median follow-up of 38 months, with respiratory failure due to aspiration pneumonia as the most common cause. Mortality was high in patients who required percutaneous endoscopic gastrostomy (7/11, 64%), and 1- year mortality was highest (31%) in those with dermatomyositis. CONCLUSION: Dysphagia is a serious and at times presenting problem in patients with inflammatory myopathy. It occurs most frequently and appears to be most refractory in patients with IBM. The mortality rate was high in patients who required percutaneous endoscopic gastrostomy, and the 1-year mortality rate was the highest in patients with dermatomyositis. PMID- 17418073 TI - Thromboangiitis obliterans (Buerger disease). PMID- 17418074 TI - Using smudge cells on routine blood smears to predict clinical outcome in chronic lymphocytic leukemia: a universally available prognostic test. AB - Recently developed prognostic tests in early Rai and Binet stage chronic lymphocytic leukemia (CLL) require considerable technologic expertise and are not available worldwide. Smudge cells are CLL cells ruptured during smear preparation. We hypothesized that smudge cell formation is inversely correlated with expression of vimentin, a cytoskeletal protein and prognostic marker, and that the percentage of smudge cells would predict prognosis in CLL. We reviewed the blood smears of 75 patients with previously untreated early and intermediate stage CLL (Rai stage 0-II) who were seen at the Mayo Clinic in Rochester, Minn, between September 1989 and December 2000. A total of 200 lymphocytes and smudge cells were counted on each slide and the results expressed as a percentage of the total lymphocytes (intact and smudged). The median percentage of smudge cells was 27% (range, 4%-72%). The percentage of smudge cells inversely correlated with vimentin expression (r=-0.57; P=.007). The median percentage of smudge cells was higher in patients with the mutated immunoglobulin heavy chain gene than in those with the unmutated immunoglobulin heavy chain gene (31% vs 13%; P=.02). Patients with less than 30% smudge cells had a median time from diagnosis to initial treatment of 72.7 months, whereas the median time from diagnosis to initial treatment in patients with 30% or more smudge cells was not reached (P=.001). The percentage of smudge cells as a continuous variable correlated with overall survival (P=.04). The estimation of smudge cells on a blood smear could be a universally available prognostic test in early-stage CLL. PMID- 17418075 TI - A profile of pedophilia: definition, characteristics of offenders, recidivism, treatment outcomes, and forensic issues. AB - Pedophilia has become a topic of increased interest, awareness, and concern for both the medical community and the public at large. Increased media exposure, new sexual offender disclosure laws, Web sites that list the names and addresses of convicted sexual offenders, politicians taking a 'get tough' stance on sexual offenders, and increased investigations of sexual acts with children have increased public awareness about pedophilia. Because of this increased awareness, it is important for physicians to understand pedophilia, its rate of occurrence, and the characteristics of pedophiles and sexually abused children. In this article, we address research that defines the various types and categories of pedophilia, review available federal data on child molestation and pornography, and briefly discuss the theories on what makes an individual develop a sexual orientation toward children. This article also examines how researchers determine if someone is a pedophile, potential treatments for pedophiles and sexually abused children, the risk of additional sexual offenses, the effect of mandatory reporting laws on both physicians and pedophiles, and limitations of the current pedophilic literature. PMID- 17418076 TI - Splanchnic artery aneurysms. AB - Autopsy studies suggest that splanchnic artery aneurysms may be more frequent than abdominal aortic aneurysms. These aneurysms are important to recognize because up to 25% may be complicated by rupture, and the mortality rate after rupture is between 25% and 70%. However, little is known about the natural history and clinical presentation of splanchnic artery aneurysms. Splenic artery aneurysms are the most common of the splanchnic artery aneurysms; multiple aneurysms are present in approximately one third of patients. Hepatic artery pseudoaneurysms are more common than true aneurysms because of increasing numbers of hepatobiliary interventional procedures. The diagnosis of splanchnic artery aneurysm should be considered in any patient with abdominal pain, a pulsatile mass, or an abdominal bruit with or without associated bleeding. However, most aneurysms are asymptomatic and are detected incidentally on imaging studies. Treatment, which can be either surgical or interventional radiology-based, should be considered in all patients with symptoms related to the aneurysms, if the aneurysm is more than 2 cm in diameter, if the patient is pregnant, or if there is demonstrated growth of the aneurysm. PMID- 17418077 TI - Clinicians' guide to new tools and features of PubMed. AB - Practicing clinicians need to have the skills required to obtain up- to-date medical information to address both the expansion of scientific knowledge and patients' increasing use of the Internet. PubMed (www.pubmed.gov) allows clinicians free access to the largest biomedical resource available. This article is the third in a Mayo Clinic Proceedings series designed specifically to help clinicians unlock the tools and information available through this valuable resource. PMID- 17418078 TI - 86-year-old woman with abdominal pain and diarrhea. PMID- 17418079 TI - Malignant melanoma in the 21st century, part 2: staging, prognosis, and treatment. AB - Critical to the clinical management of a patient with malignant melanoma is an understanding of its natural history. As with most malignant disorders, prognosis is highly dependent on the clinical stage (extent of tumor burden) at the time of diagnosis. The patient's clinical stage at diagnosis dictates selection of therapy. We review the state of the art in melanoma staging, prognosis, and therapy. Substantial progress has been made in this regard during the past 2 decades. This progress is primarily reflected in the development of sentinel lymph node biopsies as a means of reducing the morbidity associated with regional lymph node dissection, increased understanding of the role of neoangiogenesis in the natural history of melanoma and its potential as a treatment target, and emergence of innovative multimodal therapeutic strategies, resulting in significant objective response rates in a disease commonly believed to be drug resistant. Although much work remains to be done to improve the survival of patients with melanoma, clinically meaningful results seem within reach. PMID- 17418081 TI - Did a flawed study design affect the results of the VYTAL study? PMID- 17418082 TI - Sorafenib-induced pancreatitis. PMID- 17418083 TI - Impact of the FDA black box warning on physician antidepressant prescribing and practice patterns: opening Pandora's suicide box. PMID- 17418085 TI - Use of bisphosphonates in multiple myeloma: IMWG response to Mayo Clinic consensus statement. PMID- 17418086 TI - Assembly of recently translated full-length and C-terminal truncated human gamma globin chains with a pool of alpha-globin chains to form Hb F in a cell-free system. AB - Assembly of alpha-globin with translated, full-length and C-terminal truncated human gamma-globin to form Hb F was assessed in a cell-free transcription/translation system. Polysome profiles showed two amino acid C terminal-truncated gamma-chains retained on polysomes can assemble with unlabeled holo alpha-chains only after puromycin-induced chain release. Two amino acid C terminal truncated gamma-chains encoded from vectors containing a stop codon at the translation termination site were released from polysomes and assembled with alpha-chains in the absence of puromycin addition, while removal of 11 or more amino acids from the gamma-chain carboxy-terminus inhibited assembly with alpha chains. These results suggest that amino acids in the HC- and H-helix gamma-chain regions including amino acids 135-144 at the C-terminus in the translated gamma chains play a key role in assembly with alpha-chains, and that assembly occurs soon after exit of translated gamma-chains from the ribosome tunnel and release from polysomes thereby preventing stable gamma(2) homo-dimer formation. PMID- 17418087 TI - Redox potential of chlorophyll d in vitro. AB - Chlorophyll (Chl) d is a major chlorophyll in a novel oxygenic prokaryote Acaryochloris marina. Here we first report the redox potential of Chl d in vitro. The oxidation potential of Chl d was +0.88 V vs. SHE in acetonitrile; the value was higher than that of Chl a (+0.81 V) and lower than that of Chl b (+0.94 V). The oxidation potential order, Chl b>Chl d>Chl a, can be explained by inductive effect of substituent groups on the conjugated pi-electron system on the macrocycle. Corresponding pheophytins showed the same order; Phe b (+1.25 V)>Phe d (+1.21 V)>Phe a (+1.14 V), but the values were significantly higher than those of Chls, which are rationalized in terms of an electron density decrease in the pi-system by the replacement of magnesium with more electronegative hydrogen. Consequently, oxidation potential of Chl a was found to be the lowest among Chls and Phes. The results will help us to broaden our views on photosystems in A. marina. PMID- 17418088 TI - Phosphoenolpyruvate metabolism in Jerusalem artichoke mitochondria. AB - We report here initial studies on phosphoenolpyruvate metabolism in coupled mitochondria isolated from Jerusalem artichoke tubers. It was found that: (1) phosphoenolpyruvate can be metabolized by Jerusalem artichoke mitochondria by virtue of the presence of the mitochondrial pyruvate kinase, shown both immunologically and functionally, located in the inner mitochondrial compartments and distinct from the cytosolic pyruvate kinase as shown by the different pH and inhibition profiles. (2) Jerusalem artichoke mitochondria can take up externally added phosphoenolpyruvate in a proton compensated manner, in a carrier-mediated process which was investigated by measuring fluorimetrically the oxidation of intramitochondrial pyridine nucleotide which occurs as a result of phosphoenolpyruvate uptake and alternative oxidase activation. (3) The addition of phosphoenolpyruvate causes pyruvate and ATP production, as monitored via HPLC, with their efflux into the extramitochondrial phase investigated fluorimetrically. Such an efflux occurs via the putative phosphoenolpyruvate/pyruvate and phosphoenolpyruvate/ATP antiporters, which differ from each other and from the pyruvate and the adenine nucleotide carriers, in the light of the different sensitivity to non-penetrant compounds. These carriers were shown to regulate the rate of efflux of both pyruvate and ATP. The appearance of citrate and oxaloacetate outside mitochondria was also found as a result of phosphoenolpyruvate addition. PMID- 17418089 TI - Amyloid beta-protein and lipid metabolism. AB - Lipids play an important part as risk or protective factors for Alzheimer's disease. This review summarizes the current findings in which lipids influence Alzheimer's disease and introduces the molecular mechanism how these lipids are linked to amyloid production. Besides the pathological impact of amyloid in Alzheimer's disease, amyloid has a physiological function in regulating lipid homeostasis in return. The understanding of the resulting regulatory cycles between amyloid precursor protein processing and lipids provides a platform for the development of new causal therapeutic approaches for Alzheimer's disease. PMID- 17418090 TI - Metacognitive therapy for obsessive-compulsive disorder: a case series. AB - Cognitive-behavior therapy and exposure and response prevention are the most effective psychological treatments for obsessive-compulsive disorder (OCD). However, these approaches often produce variable results with the majority of treated individuals remaining symptomatic. This study evaluated a new form of cognitive therapy based on Wells' metacognitive model of OCD. Treatment efficacy was assessed using single case methodology in 4 consecutively referred individuals. At post-treatment, all participants made clinically significant change on a range of standardized outcome measures and these gains were largely maintained through to 6-month follow-up. Metacognitive therapy could be an effective and time efficient treatment for OCD. PMID- 17418091 TI - Axin-independent phosphorylation of APC controls beta-catenin signaling via cytoplasmic retention of beta-catenin. AB - It has been shown that accumulation of free beta-catenin leads to mobility shift of adenomatous polyposis coli (APC) protein and that Axin facilitates this process. Here we show that the beta-catenin-mediated mobility shift of APC is due to phosphorylation of two domains of APC by casein kinase 1epsilon/glycogen synthase kinase 3beta and unknown kinase(s), respectively. Interestingly, our results suggest that this process does not require Axin. The phosphorylated APC showed higher affinity to beta-catenin in vivo, and fragments of APC containing the phosphorylated domains can inhibit beta-catenin/Tcf-mediated reporter activity regardless of their ability to reduce the level of beta-catenin. From our data we propose a new role of APC: accumulation of excessive cytoplasmic beta catenin induces phosphorylation of APC and the phosphorylated APC retains beta catenin in cytoplasm to prevent excessive beta-catenin signaling. The retained beta-catenin in cytoplasm by APC may be down-regulated by Axin 2, which is induced by beta-catenin/Tcf signaling. PMID- 17418092 TI - Hypertrophy and atrophy inversely regulate Caveolin-3 expression in myoblasts. AB - Caveolin-3 (Cav-3) is a muscle-specific membrane protein crucial for myoblast differentiation, as loss of the protein due to mutations within the gene causes an autosomal dominant form of limb girdle muscular dystrophy 1-c. Here we show that along with p38 activity the PI3-kinase/AKT/mTOR pathway is required for proper Cav-3 up-regulation during muscle differentiation and hypertrophy, as confirmed by the marked increase of Cav-3 expression in hypertrophied C2C12 cells transfected with an activated form of AKT. Accordingly, Cav-3 expression was further increased during hypertrophy of L6C5 myoblasts treated with Arg(8) vasopressin and in hypertrophic muscles of MLC/mIGF-1 transgenic mice. In contrast, Cav-3 expression was down-regulated in C2C12 myotubes exposed to atrophic stimuli such as starvation or treatment with dexamethasone. This study clearly suggests that Cav-3 expression is causally linked to the maturation of muscle phenotype and it is tightly regulated by hypertrophic and atrophic stimuli. PMID- 17418093 TI - Development of a disease-specific model to evaluate endothelial dysfunction in patients with diabetes mellitus. AB - Diabetic patients have an increased cardiovascular risk. We propose to characterize the endothelial dysfunction in a disease-specific in vitro model. Human saphenous vein endothelial cells (HSVEC) were isolated from coronary artery bypass patients without and with non-insulin-dependent diabetes mellitus. Growth kinetics and proinflammatory responses (expression of adhesion molecules, cytokines) were documented under non-stimulating conditions. Diabetic HSVEC showed delayed growth kinetics with reduced cell densities of about 40%. During exponential growth of diabetic EC, the surface expression of adhesion molecules was increased 10-fold (p< or =0.05). However, in a monolayer the expression adapted to low levels of non-diabetic EC. In addition, diabetic EC produced significantly more soluble E-selectin, VCAM-1, IL-6 and MCP-1. Our results suggest a link between the pathologically proinflammatory basic state of diabetic EC and the endothelial dysfunction in diabetic disease. Therefore, this in vitro model could be used for investigating early dysfunction and environmental effects on pathological endothelium. PMID- 17418094 TI - Regulation of MDR1 gene expression in multidrug-resistant cancer cells is independent from YB-1. AB - The MDR1 gene encoded transmembrane ABC-transporter MDR1/P-glycoprotein can mediate the phenotype of multidrug resistance (MDR), a major obstacle in the clinical management of cancer patients. It was hypothesized that YB-1 is a fundamental regulatory factor of the MDR1 gene in tumor cells and can therewith enhance drug resistance. To analyze the potential impact of YB-1 in MDR cancer cells, two specific anti-YB-1 small interfering RNAs (siRNAs) were designed for transient triggering the gene-silencing RNA interference (RNAi) pathway in the MDR cell lines EPG85-257RDB and EPP85-181RDB as well as in their drug-sensitive counterparts EPG85-257P and EPP85-181P. Since both siRNAs showed biological activity, for stable inhibition of YB-1 corresponding tetracycline-inducible short hairpin RNA (shRNA)-encoding expression vectors were designed. By treatment of the cancer cells with these constructs, the expression of the targeted YB-1 encoding mRNA and protein was completely inhibited following tetracycline exposure. These gene-silencing effects were not accompanied by modulation of the MDR1 expression or by reversal of the drug-resistant phenotype. In conclusion, the data demonstrate the utility of the analyzed RNAs as powerful laboratory tools and indicate that YB-1 is not involved in the regulation of the MDR1 gene or the development of the drug-resistant phenotype in MDR cancer cells. PMID- 17418095 TI - Actin binding and proline rich motifs of CR16 play redundant role in growth of vrp1Delta cells. AB - CR16, (Glucocorticoid-regulated) belongs to the verprolin family of proteins which are characterized by the presence of a V domain (verprolin) at the N terminal. Expression of CR16 suppressed the growth and endocytosis defect of vrp1Delta strain without correcting the actin patch polarization defect. The V domain of CR16 is critical for suppression of the growth defect of vrp1Delta strain but not for localisation to cortical actin patches. Mutations in the actin binding motif alone did not abolish the activity of CR16 but the mutations in combination with deletion of N-terminal proline rich motif abolished the ability of CR16 to suppress the growth defect. This suggests that the V domain of CR16 has two functionally redundant motifs and either one of these motifs is sufficient for suppressing the growth defect of vrp1Delta strain. This is in contrast to the observation that both WIP and WIRE require the actin binding motif for their activity. PMID- 17418096 TI - Metformin prevents methylglyoxal-induced apoptosis of mouse Schwann cells. AB - Methylglyoxal (MG) is involved in the pathogenesis of diabetic complications via the formation of advanced glycation end products (AGEs) and reactive oxygen species (ROS). To clarify whether the antidiabetic drug metformin prevents Schwann cell damage induced by MG, we cultured mouse Schwann cells in the presence of MG and metformin. Cell apoptosis was evaluated using Hoechst 33342 nuclear staining, caspase-3 activity, and c-Jun-N-terminal kinase (JNK) phosphorylation. Intracellular ROS formation was determined by flow cytometry, and AMP-activated kinase (AMPK) phosphorylation was also examined. MG treatment resulted in blunted cell proliferation, an increase in the number of apoptotic cells, and the activation of caspase-3 and JNK along with enhanced intracellular ROS formation. All of these changes were significantly inhibited by metformin. No significant activation of AMPK by MG or metformin was observed. Taken together, metformin likely prevents MG-induced apoptotic signals in mouse Schwann cells by inhibiting the formation of AGEs and ROS. PMID- 17418097 TI - Obestatin reduces food intake and suppresses body weight gain in rodents. AB - Obestatin was recently described as a bioactive peptide encoded for by the same gene as ghrelin but with opposite actions on food intake. Although some groups have confirmed these findings others find no effect. We investigated the effect of obestatin on feeding in rodents over a wide range of doses. Acute administration of obestatin inhibited feeding at doses of 10-100 nmol/kg i.p. in mice and 100-300 nmol/kg i.p. in lean and Zucker fatty rats. Interestingly, the dose-response relationship was U-shaped such that both low and high doses were without effect in either species. Treatment of mice with obestatin over a 7-day period decreased body weight gain and food consumption. Overall, obestatin suppressed food intake and body weight gain in rodent and an unusual dose response relationship was found. These findings may explain the difficulties in reproducing the effects of obestatin on feeding reported by some groups. PMID- 17418098 TI - Ligand-dependent transcription of estrogen receptor alpha is mediated by the ubiquitin ligase EFP. AB - Estrogen-mediated ubiquitylation and subsequent degradation of the estrogen receptor alpha (ERalpha) appears to be involved in the transcriptional activity of ERalpha. We show that the estrogen-responsive finger protein (EFP) interacts with and ubiquitylates ERalpha. EFP promoted the ubiquitylation of ERalphain vitro and in vivo and consequently promoted the degradation of ERalpha. The interaction between EFP and ERalpha was greatly enhanced in the presence of estrogen. The action of EFP on ERalpha in the presence of estrogen resulted in a robust interaction between ERalpha and Tip60, one of the transcriptional coactivators, leading to activation of ERalpha transcriptional activity. However, a dominant negative mutant of EFP lacking the RING domain prolonged the half-life of ERalpha and inhibited the transcription by ERalpha. Our results indicate that EFP functions as a cofactor for ERalpha-mediated transcription, thus suggesting that ERalpha-mediated transcription is closely linked to the ubiquitylation of ERalpha. PMID- 17418099 TI - Estrogen-related receptor alpha is essential for the expression of antioxidant protection genes and mitochondrial function. AB - Estrogen-related receptor alpha (ERRalpha) is an important mediator of mitochondrial biogenesis and function. To investigate the transcriptional network controlling these phenomena, we investigated mitochondrial gene expression in embryonic fibroblasts isolated from ERRalpha null mice. Peroxisome proliferator activated receptor gamma coactivator-1alpha (PGC-1alpha) stimulated mitochondrial gene expression program in control cells, but not in the ERRalpha null cells. Interestingly, the induction of levels of mitochondrial oxidative stress protection genes in response to increased PGC-1alpha levels was dependent on ERRalpha. Furthermore, we found that the PGC-1alpha-mediated induction of estrogen-related receptor gamma and nuclear respiratory factor 2 (NRF-2), was dependent on the presence of ERRalpha. Basal levels of NRF-2 were decreased in the absence of ERRalpha. The absence of ERRalpha resulted in a decrease in citrate synthase enzyme activity in response to PGC-1alpha overexpression. Our results indicate an essential role for ERRalpha as a key regulator of oxidative metabolism. PMID- 17418100 TI - HTLV-1 Tax-induced NFkappaB activation is independent of Lys-63-linked-type polyubiquitination. AB - Human T-cell leukemia virus type 1 (HTLV-1) Tax-induced activation of nuclear factor-kappaB (NFkappaB) is thought to play a critical role in T-cell transformation and onset of adult T-cell leukemia. However, the molecular mechanism of the Tax-induced NFkappaB activation remains unknown. One of the mitogen-activated protein kinase kinase kinses (MAP3Ks) members, TAK1, plays a critical role in cytokine-induced activation of NFkappaB, which involves lysine 63-linked (K63) polyubiquitination of NEMO, a noncatalytic subunit of the IkappaB kinase complex. Here we show that Tax induces K63 polyubiquitination of NEMO. However, TAK1 is dispensable for Tax-induced NFkappaB activation, and deubiquitination of the K63 polyubiquitin chain failed to block Tax-induced NFkappaB activation. In addition, silencing of other MAP3Ks, including MEKK1, MEKK3, NIK, and TPL-2, did not affect Tax-induced NFkappaB activation. These results strongly suggest that unlike cytokine signaling, Tax-induced NFkappaB activation does not involve K63 polyubiquitination-mediated MAP3K activation. PMID- 17418101 TI - Lack of sphingosine 1-phosphate-degrading enzymes in erythrocytes. AB - Platelets are known to store a large amount of the bioactive lipid molecule sphingosine 1-phosphate (S1P) and to release it into the plasma in a stimuli dependent manner. Erythrocytes can also release S1P, independently from any stimuli. We measured the S1P and sphingosine (Sph) levels in erythrocytes by HPLC and found that the contribution of erythrocyte S1P to whole blood S1P levels is actually higher than that of platelets. In vitro assays demonstrated that erythrocytes possess much weaker Sph kinase activity compared to platelets but lack the S1P-degrading activities of either S1P lyase or S1P phosphohydrolase. This combination may enable erythrocytes to maintain a high S1P content relative to Sph. The absence of both S1P-degrading enzymes has not been reported for other cell types. Thus, erythrocytes may be specialized cells for storing and supplying plasma S1P. PMID- 17418102 TI - 15-deoxy-Delta12,14-prostaglandin J2-induced down-regulation of endothelial nitric oxide synthase in association with HSP70 induction. AB - A natural ligand of peroxisome proliferator-activated receptor gamma (PPARgamma), 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), decreases endothelial nitric oxide synthase (eNOS) expression by an unknown mechanism. Here we found that 15d-PGJ(2)-induced eNOS reduction is inversely associated with heat shock protein 70 (HSP70) induction in endothelial cells. Treatment of cells with 15d PGJ(2) decreased eNOS protein expression in a concentration- and time-dependent manner, but independently of PPARgamma with no effect on mRNA levels. Although 15d-PGJ(2) elicited endothelial apoptosis, inhibition of both pan-caspases and cathepsins failed to reverse reduction of eNOS protein. Interestingly, we observed that 15d-PGJ(2) induced HSP70 in a dose-dependent manner. Immunoprecipitation and heat shock treatment demonstrated that eNOS reduction was strongly related to HSP70 induction. Cellular fractionation revealed that treatment with 15d-PGJ(2) increased eNOS distribution 2.5-fold from soluble to insoluble fractions. These findings provide new insights into mechanisms whereby eNOS regulation by 15d-PGJ(2) is related to HSP70 induction. PMID- 17418103 TI - Lysophosphatidic acid induces osteocyte dendrite outgrowth. AB - Osteocytes elaborate an extensive mechanosensory network in bone matrix and communicate intercellularly via gap junctions established at dendrite termini. We developed a method to measure osteocyte dendritogenesis in vitro using a modified transwell assay and determined that the lipid growth factor lysophosphatidic acid (LPA) is a potent stimulator of dendrite outgrowth in MLO-Y4 osteocytes. The stimulatory effects were dose-dependent with maximal outgrowth observed within a physiological range of LPA. LPA-treated osteocytes exhibited distinct rearrangements of the actin cytoskeleton and a more stellate morphology than control cells. LPA also promoted osteocyte chemotaxis, suggesting a shared molecular mechanism between dendrite outgrowth and cell motility. The LPA-induced increase in dendrite formation was blocked by the specific LPA-receptor antagonist Ki16425 and by pertussis toxin. Bone cells in vivo encounter platelet derived LPA in regions of bone damage, and we postulate that this lipid factor is important for re-establishing osteocyte connectivity during fracture repair. PMID- 17418104 TI - Insulin and amino acid availability regulate atrogin-1 in avian QT6 cells. AB - New evidence has demonstrated that the expression of major genes, termed atrogenes, controls the ubiquitin-proteasome proteolytic pathway. The present work aimed to study the impact of insulin and amino acids on the expression of one of these atrogenes, the E3 ubiquitin ligase Muscle Atrophy F box (MAFbx, also called atrogin-1), in quail muscle (QT6) fibroblasts. First, we characterized atrogin-1 in QT6 cells and demonstrated the insulin sensitivity of these cells. Second, we showed that insulin reduced atrogin-1 mRNA via the phosphatidylinositol-3'kinase (PI3K)/protein kinase B (PKB or AKT)/target of rapamycin (TOR) pathway. Atrogin-1 expression also depended on the availability of an individual amino acid, i.e., methionine. Moreover, the amino acid-induced reduction of atrogin-1 was inhibited by rapamycin, indicating the involvement of the TOR pathway in such regulation. In conclusion, expression of the ubiquitin ligase atrogin-1 is regulated by both insulin and amino acids through the TOR pathway. PMID- 17418105 TI - Mutant presenilin 2 increased oxidative stress and p53 expression in neuronal cells. AB - The learning and memory impairment of presenilin 2 transgenic mice was mentioned previously. In this study, exposing the presenilin 2 transfected PC12 cells to the 50 microM Abeta(25-35), 30 mM l-glutamate and 50 microM H(2)O(2) resulted in significant increase 8-oxodG and p53 levels of the cells expressing the mutant gene. The increase was also found in the mutant presenilin 2 transgenic mice brains age-dependently in comparison to that in the wild-type presenilin 2 transgenic mice and non-transgenic ones. These findings indicated that mutant presenilin 2 clearly increases oxidative stress and p53 expression, which could be implicated in promoting mutant presenilin 2-induced neurodegeneration in Alzheimer's disease, and the influence of mutant presenilin 2 in Alzheimer's disease may be brain regional and age related effects. PMID- 17418106 TI - Go but not Gi2 or Gi3 is required for muscarinic regulation of heart rate and heart rate variability in mice. AB - Muscarinic receptor-mediated cardiac parasympathetic activity is essential for regulating heart rate and heart rate variability (HRV). It has not been clear which G(i)/G(o) protein is responsible for these effects. We addressed this question using knockout mice that lack G protein alpha(i2), alpha(i3), or alpha(o) specifically. Unlike previously reported, our alpha(o)-null mice had significantly more survivors with normal life span. Isolated hearts from alpha(o) null mice demonstrated much less sensitivity to the negative chronotropic effects of the muscarinic agonist carbachol to lower heart rate at baseline and a more profound effect under the stimulation of the beta-adrenergic agonist isoproterenol. In the presence of parasympathetic activation indirectly produced by methoxamine, an alpha(1)-adrenergic agonist, alpha(o)-null mice showed markedly decreased HRV compared with wild-type control mice. These differences in heart rate and HRV were not observed in alpha(i2)-null or alpha(i3)-null mice. Our findings establish an essential role for alpha(o) G protein in the anti adrenergic effect of carbachol on heart rate regulation. PMID- 17418107 TI - Molecular characterization of GDD1/TMEM16E, the gene product responsible for autosomal dominant gnathodiaphyseal dysplasia. AB - The human GDD1/TMEM16E gene has been found to be mutated in gnathodiaphyseal dysplasia, an unusual skeletal syndrome with autosomal dominant inheritance. The molecular and biochemical function(s) of GDD1 protein has not yet been elucidated. In this study, we examined the murine GDD1 gene expression pattern during embryonic development, and characterized the cellular and tissue localizations of its gene product using a GDD1-specific antibody. In the developing embryos, GDD1 mRNA expression was principally associated with differentiating and developing somites, with a highly complex spatiotemporal pattern that involved the myotomal and sclerotomal lineages of somites. Biochemical studies indicated that GDD1 protein is an integral membrane glycoprotein that resides predominantly in intracellular vesicles. Immunohistochemical analysis showed a high level of murine GDD1 protein expression in cardiac and skeletal muscle tissues, and in growth-plate chondrocytes and osteoblasts in bone. These observations suggest diverse cellular role(s) of GDD1 in the development of musculoskeletal system. PMID- 17418108 TI - The combination of peripheral nerve grafts and acidic fibroblast growth factor enhances arginase I and polyamine spermine expression in transected rat spinal cords. AB - Treatment with a combination of peripheral nerve grafts and acidic fibroblast growth factor improves hind limb locomotor function after spinal cord transection. This study examined the effect of treatment on expression of arginase I (Arg I) and polyamines. Arg I expression was low in the spinal cords of normal rats but increased following spinal injury. Only fully repaired spinal cords expressed higher Arg I levels 6-14 days following repair. In 10-day repaired spinal cords, high Arg I immunoreactivity was detected in motoneurons and alternatively activated macrophages in the graft area and graft-stump edges, and high levels of the polyamine spermine were expressed by macrophages within the intercostal nerve graft. Thus, in addition to enhancing the expression of Arg I and spermine in repaired spinal cords, our treatment may recruit activated macrophages and create a more favorable environment for axonal regrowth. PMID- 17418109 TI - Early cell death in the brain of fetal preterm lambs after hypoxic-ischemic injury. AB - The objective of the present study was to evaluate using premature fetal lambs the effect of cerebral hypoxia-ischemia induced by partial occlusion of the umbilical cord on the type of cell death which occurs in different brain regions and to ascertain some of the neural pathways which may underlie the associated pathologies. Lambs were sacrificed either immediately after a 1 h hypoxic ischemic insult or 3 h later. Brains were fixed by perfusion and blocks of the different brain territories were processed for light microscopy (hematoxylin eosin, Nissl staining), electron transmission microscopy and quantification of apoptosis by the TUNEL method. Other fixed brains were dissociated and labeled by nonyl acridine orange to determine mitochondrial integrity. Non-fixed brains were also used for membrane asymmetry studies, in which cell suspensions were analyzed by flow cytometry to quantify apoptosis. In both hypoxic-ischemic groups, necrotic-like neurons were observed mainly in the mesencephalon, pons, deep cerebellar nuclei and basal nuclei, whereas apoptotic cells were extensively found both in white and gray matter and were not limited to regions where necrotic neurons were present. The 3 h post-partial cord occlusion group, but not the 0 h group, showed a generalized alteration of cell membrane asymmetry and mitochondrial integrity as revealed by Annexin V/PI flow cytometry and nonyl acridine orange studies, respectively. Our results show that the apoptotic/necrotic patterns of cell death occurring early after hypoxic-ischemic injury are brain-region-specific and have distinct dynamics and suggest that therapeutic strategies aimed at rescuing cells from the effects of hypoxia/ischemia should be aimed at blocking the apoptotic components of brain damage. PMID- 17418110 TI - Evidence for elevated nicotine-induced structural plasticity in nucleus accumbens of adolescent rats. AB - Male Long-Evans rats were administered nicotine bitartrate or sodium tartrate either during adolescence (p29-43) or adulthood (p80-94). Route of administration was via subcutaneously implanted osmotic pump (initial dose 2.0 mg/kg/day, free base). Five weeks following nicotine administration, brains were processed for Golgi-Cox staining. Medium spiny neurons from nucleus accumbens (NAc) shell were digitally reconstructed for morphometric analysis. Total dendritic length and branch number were greater in medium spiny neurons from animals pretreated with nicotine during adolescence. A branch order analysis indicated that increased branch number was specific to higher order branches. Mean branch lengths did not differ with respect to treatment as a function of branch order. Thus, nicotine induced increases in total dendritic length were a function of greater numbers of branches, not increased segment length. In contrast, adult nicotine exposure did not significantly alter total dendritic length or branch number of medium spiny neurons. Total dendritic length and branch number of a second morphological type, the large aspiny neuron, did not differ following either adolescent or adult pretreatment. The age-dependent alteration of accumbal structure was associated with qualitatively different behavioral responses to drug challenge. These data provide evidence that drug-induced structural plasticity in nucleus accumbens is considerably more pronounced during adolescence. PMID- 17418111 TI - Biochemical markers of oxidative stress in Perna viridis exposed to mercury and temperature. AB - Oxidative damage and antioxidant properties have been studied in Perna viridis subjected to short-term exposure to Hg along with temperature (72h) and long-term temperature exposures (14 days) as pollution biomarkers. The elevated thiobarbituric acid reactive substances (TBA-RS) levels observed in gills and digestive gland under exposure to Hg, individually and combined with temperature, as also long-term temperature stress have been assigned to the oxidative damage resulting in lipid peroxidation (LPX). Increased activities of antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione-S-transferase (GST) both in gills and digestive glands under long-term exposures to temperatures are more prominent to heat rather than cold stress suggesting activation of physiological mechanism to scavenge the ROS produced during heat stress. Also decreased values of reduced glutathione (GSH) on long exposures to temperature stress indicate utilisation of this antioxidant, either to scavenge oxiradicals or act in combination with other enzymes, was more than its production capacity under heat stress. The results suggest that temperature variation does alter the active oxygen metabolism by modulating antioxidant enzyme activities, which can be used as biomarker to detect sublethal effects of pollution. PMID- 17418112 TI - Inflexibility of experts--reality or myth? Quantifying the Einstellung effect in chess masters. AB - How does the knowledge of experts affect their behaviour in situations that require unusual methods of dealing? One possibility, loosely originating in research on creativity and skill acquisition, is that an increase in expertise can lead to inflexibility of thought due to automation of procedures. Yet another possibility, based on expertise research, is that experts' knowledge leads to flexibility of thought. We tested these two possibilities in a series of experiments using the Einstellung (set) effect paradigm. Chess players tried to solve problems that had both a familiar but non-optimal solution and a better but less familiar one. The more familiar solution induced the Einstellung (set) effect even in experts, preventing them from finding the optimal solution. The presence of the non-optimal solution reduced experts' problem solving ability was reduced to about that of players three standard deviations lower in skill level by the presence of the non-optimal solution. Inflexibility of thought induced by prior knowledge (i.e., the blocking effect of the familiar solution) was shown by experts but the more expert they were, the less prone they were to the effect. Inflexibility of experts is both reality and myth. But the greater the level of expertise, the more of a myth it becomes. PMID- 17418113 TI - nanos1 is required to maintain oocyte production in adult zebrafish. AB - Development of the germline requires the specification and survival of primordial germ cells (PGCs) in the embryo as well as the maintenance of gamete production during the reproductive life of the adult. These processes appear to be fundamental to all Metazoans, and some components of the genetic pathway regulating germ cell development and function are evolutionarily conserved. In both vertebrates and invertebrates, nanos-related genes, which encode RNA-binding zinc finger proteins, have been shown to play essential and conserved roles during germ cell formation. In Drosophila, maternally supplied nanos is required for survival of PGCs in the embryo, while in adults, nanos is required for the continued production of oocytes by maintaining germline stem cells self-renewal. In mice and zebrafish, nanos orthologs are required for PGC survival during embryogenesis, but a role in adults has not been explored. We show here that nanos1 in zebrafish is expressed in early stage oocytes in the adult female germline. We have identified a mutation in nanos1 using a reverse genetics method and show that young female nanos mutants contain oocytes, but fail to maintain oocyte production. This progressive loss of fertility in homozygous females is not a phenotype that has been described previously in the zebrafish and underlines the value of a reverse genetics approach in this model system. PMID- 17418114 TI - A GATA/RUNX cis-regulatory module couples Drosophila blood cell commitment and differentiation into crystal cells. AB - Members of the RUNX and GATA transcription factor families play critical roles during hematopoiesis from Drosophila to mammals. In Drosophila, the formation of the crystal cell hematopoietic lineage depends on the continuous expression of the lineage-specific RUNX factor Lozenge (Lz) and on its interaction with the GATA factor Serpent (Srp). Crystal cells are the main source of prophenoloxidases (proPOs), the enzymes required for melanization. By analyzing the promoter regions of several insect proPOs, we identify a conserved GATA/RUNX cis regulatory module that ensures the crystal cell-specific expression of the three Drosophila melanogaster proPO. We demonstrate that activation of this module requires the direct binding of both Srp and Lz. Interestingly, a similar GATA/RUNX signature is over-represented in crystal cell differentiation markers, allowing us to identify new Srp/Lz target genes by genome-wide screening of Drosophila promoter regions. Finally, we show that the expression of lz in the crystal cells also relies on Srp/Lz-mediated activation via a similar module, indicating that crystal cell fate choice maintenance and activation of the differentiation program are coupled. Based on our observations, we propose that this GATA/RUNX cis-regulatory module may be reiteratively used during hematopoietic development through evolution. PMID- 17418115 TI - Molecular patterning mechanism underlying metamorphosis of the thoracic leg in Manduca sexta. AB - The tobacco hornworm Manduca sexta, like many holometabolous insects, makes two versions of its thoracic legs. The simple legs of the larva are formed during embryogenesis, but then are transformed into the more complex adult legs at metamorphosis. To elucidate the molecular patterning mechanism underlying this biphasic development, we examined the expression patterns of five genes known to be involved in patterning the proximal-distal axis in insect legs. In the developing larval leg of Manduca, the early patterning genes Distal-less and Extradenticle are already expressed in patterns comparable to the adult legs of other insects. In contrast, Bric-a-brac and dachshund are expressed in patterns similar to transient patterns observed during early stages of leg development in Drosophila. During metamorphosis of the leg, the two genes finally develop mature expression patterns. Our results are consistent with the hypothesis that the larval leg morphology is produced by a transient arrest in the conserved adult leg patterning process in insects. In addition, we find that, during the adult leg development, some cells in the leg express the patterning genes de novo suggesting that the remodeling of the leg involves changes in the patterning gene regulation. PMID- 17418116 TI - TGF-beta isoform signaling regulates secondary transition and mesenchymal-induced endocrine development in the embryonic mouse pancreas. AB - Transforming growth factor-beta (TGF-beta) superfamily signaling has been implicated in many developmental processes, including pancreatic development. Previous studies are conflicting with regard to an exact role for TGF-beta signaling in various aspects of pancreatic organogenesis. Here we have investigated the role of TGF-beta isoform signaling in embryonic pancreas differentiation and lineage selection. The TGF-beta isoform receptors (RI, RII and ALK1) were localized mainly to both the pancreatic epithelium and mesenchyme at early stages of development, but then with increasing age localized to the pancreatic islets and ducts. To determine the specific role of TGF-beta isoforms, we functionally inactivated TGF-beta signaling at different points in the signaling cascade. Disruption of TGF-beta signaling at the receptor level using mice overexpressing the dominant-negative TGF-beta type II receptor showed an increase in endocrine precursors and proliferating endocrine cells, with an abnormal accumulation of endocrine cells around the developing ducts of mid-late stage embryonic pancreas. This pattern suggested that TGF-beta isoform signaling may suppress the origination of secondary transition endocrine cells from the ducts. Secondly, TGF-beta isoform ligand inhibition with neutralizing antibody in pancreatic organ culture also led to an increase in the number of endocrine positive cells. Thirdly, hybrid mix-and-match in vitro recombinations of transgenic pancreatic mesenchyme and wild-type epithelium also led to increased endocrine cell differentiation, but with different patterns depending on the directionality of the epithelial-mesenchymal signaling. Together these results suggest that TGF-beta signaling is important for restraining the growth and differentiation of pancreatic epithelial cells, particularly away from the endocrine lineage. Inhibition of TGF-beta signaling in the embryonic period may thus allow pancreatic epithelial cells to progress towards the endocrine lineage unchecked, particularly as part of the secondary transition of pancreatic endocrine cell development. TGF-beta RII in the ducts and islets may normally serve to downregulate the production of beta cells from embryonic ducts. PMID- 17418117 TI - A normative value pilot study: levels of uranium in urine samples from UK civilians. AB - A normative study of the levels of urinary uranium in the general UK population is needed for comparison with levels in UK military and ex-military personnel who served where munitions containing depleted uranium (DU) were used. As preparation, this pilot study trialled the process of collecting 24-h samples from adult male civilians, and compared the measurements from 24-h samples with those from spot samples taken over the subsequent 24h. The purpose was to assess the relative utility of the two types of samples. Twenty-five convalescent hospital in-patients were recruited as participants. Uranium concentrations in the 24-h samples ranged from 1 to 10.6 ng l(-1); in the spots, from not detectable to 38.1 ng l(-1). Normalised to creatinine, concentrations in the 24h samples ranged from approximately 100 to 800 ng mol(-1) creatinine; in the spot samples, from not detectable to approximately 4000 ng mol(-1) creatinine. The ranges appear similar to those reported for residents of the US. The distribution of spot sample results indicated that 95% of a participant's creatinine-adjusted concentrations from spot samples would be within the range 40-250% of his mean. Adjusting for creatinine almost entirely eliminated a slight indication of diurnal variation in urinary uranium concentration in spot samples. All the 24-h samples and 131 out of the 133 spot samples showed ratios of isotopes (238)U to (235)U consistent with natural uranium (i.e. neither enriched nor depleted). Slightly elevated ratios in two spot samples were not supported by other samples from the same participants, indicating that slightly elevated ratios may be recorded on very low concentration (<1 ng l(-1)) samples. In the main, quantification of this isotope ratio from spot samples was only slightly more variable than from 24-h samples. Complete 24-h urine samples gave better precision than spot samples in estimating uranium concentrations at these low levels, but presented more logistic difficulties in the collection of the samples. Clarification of the relative merits of alternative sampling strategies enables the design of a wider study to be optimised. PMID- 17418118 TI - Involvement of phosphatases in the anchorage-dependent regulation of ERK2 activation. AB - Activation of extracellular signal-regulated kinase (ERK) is known to be regulated by cell adhesion, namely "anchorage dependence". Most studies on the anchorage-dependent regulation have focused on the upstream activating components. We previously reported that the focal adhesion protein vinexin beta can induce the anchorage-independent activation of ERK2. We show here that vinexin beta-induced anchorage-independent activation of ERK2 involves prevention of the dephosphorylation of ERK2, but not the promotion of MEK1 or Raf1 activity. Furthermore, knockdown of vinexin beta resulted in a faster dephosphorylation of ERK2 in A549 cells. Moreover, the coexpression of MKP3/rVH6, an ERK2 specific phosphatase, suppressed the anchorage-independent activation of ERK2 induced by vinexin beta. These results suggest that vinexin beta can prevent the dephosphorylation of ERK2 stimulated by cell detachment, leading to the anchorage independent activation of ERK2. Furthermore, we found that phosphatase activity directed against activated ERK2 was higher in suspended cells than in adherent cells. In addition, orthovanadate efficiently induces anchorage-independent activation of ERK2 without marked activation of MEK1 in NIH3T3 cells. These observations suggest that the anchorage dependence of ERK1/2 activation is regulated not only by upstream kinases, Raf1 and MEK, but also by phosphatases acting against ERK1/2 and that vinexin beta can induce anchorage-independent activation of ERK by preventing the inactivation of ERK1/2. PMID- 17418119 TI - The endothelium-derived hyperpolarising factor (EDHF) in isolated bovine choroidal arteries. AB - The present study reports of an endothelium-dependent and NO- and prostanoid independent relaxation in isolated choroidal arteries, and evaluates the hypothesis of an endothelium-derived hyperpolarising factor (EDHF) playing a role in the choroidal circulation. Choroidal arteries were isolated from bovine eyes and mounted in a small vessel wire-myograph for isometric tension recording. Concentration-response curves for acetylcholine (0.1nM-10microM) were constructed in isolated choroidal arteries contracted with 10microM norepinephrine. Acetylcholine induced a concentration-dependent relaxation in the choroidal arteries. The presence of the NO-synthase inhibitor L-NA and the cyclo-oxygenase inhibitor indomethacin only had a limited effect on this relaxation. All further experiments were performed in the presence of L-NA and indomethacin, in order to study the NO- and prostanoid-independent part of the acetylcholine-relaxations. Both removal of the vascular endothelium or the presence of an increased K(+) concentration in the organ bath abolished the NO- and prostanoid-independent part of the acetylcholine-relaxations. The presence of TEA, a rather non-specific K(+) channel blocker, significantly reduced the acetylcholine-relaxations. Simultaneous application of apamin (an inhibitor of small-conductance Ca(2+) activated K(+) channels) and charybdotoxin (an inhibitor of intermediate- and large-conductance Ca(2+)-activated K(+) channels) abolished the acetylcholine induced relaxation and even resulted in a concentration-dependent contraction. Transmembrane potential recordings in isolated choroidal arteries revealed a clear membrane hyperpolarisation in the vascular smooth muscle cells of isolated choroidal arteries. It was therefore concluded that the acetylcholine-induced relaxation of choroidal arteries in the presence of NO-synthase and cyclo oxygenase inhibitors is mediated by an endothelium-derived hyperpolarising factor. This EDHF seems to be of more importance than endothelium-derived NO or prostanoids. PMID- 17418120 TI - Effect of pitavastatin on experimental choroidal neovascularization in rats. AB - The association between the use of statins and age-related macular degeneration (AMD), a leading cause of blindness, has been evaluated in many clinical studies; however, the results have been contradictory. We evaluated the effect of pitavastatin administration on laser-induced experimental choroidal neovascularization (CNV) in rats. Brown Norway rats received pitavastatin (1.0mg/kg per day) for 1day prior to laser-induced CNV and continued to receive the drug for 14days. Fluorescein angiograms were graded by masked observers. CNV area and thickness were assessed by fluorescein isothiocyanate-labeled dextran angiography and histology, respectively. Vascular endothelial growth factor (VEGF), monocyte chemoattractant protein-1 (Ccl-2; also known as MCP-1), and intercellular adhesion molecule-1 (ICAM-1) mRNA levels were measured using reverse-transcription polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR. Pitavastatin-treated rats had significantly less fluorescence leakage compared with the vehicle-treated rats estimated by CNV score using fluorescein angiography. Both the area and the thickness of CNV in pitavastatin-treated rats were significantly reduced compared with the vehicle treated rats. Gene expression of VEGF, Ccl-2, and ICAM-1 were significantly decreased by pitavastatin administration in experimental CNV. Thus, we demonstrated that the therapeutic dose of pitavastatin for human hypocholesterolemia effectively suppressed experimental CNV in rats. The use of pitavastatin may be helpful in preventing CNV development in AMD patients. PMID- 17418121 TI - Changes in the subcellular distribution of NADPH oxidase subunit p47phox in dendrites of rat dorsomedial nucleus tractus solitarius neurons in response to chronic administration of hypertensive agents. AB - NADPH oxidase-generated superoxide can modulate crucial intracellular signaling cascades in neurons of the nucleus tractus solitarius (NTS), a brain region that plays an important role in cardiovascular processes. Modulation of NTS signaling by superoxide may be linked to the subcellular location of the mobile NADPH oxidase p47(phox) subunit, which is known to be present in dendrites of NTS neurons. It is not known, however, if hypertension can produce changes in the trafficking of p47(phox) in defined NTS subregions, particularly the preferentially barosensitive dorsomedial NTS (dmNTS), or preferentially gastrointestinal medial NTS (mNTS). We used immunogold electron microscopy to determine if p47(phox) localization was differentially affected in dendritic profiles of neurons from these NTS subregions of the rat in response to distinct models of hypertension, namely chronic 7-day subcutaneous administration of angiotensin II (AngII), or phenylephrine. In small (<1 microm) dendritic processes, both AngII and phenylephrine produced a decrease in intracellular p47(phox) labeling selectively in dmNTS neurons. In intermediate-size (1-2 microm) dendritic profiles in the dmNTS region only, there was an increase in p47(phox) labeling in response to each hypertensive agent, although these changes occurred in different subcellular compartments. There was an increase in non vesicular labeling in response to AngII, but an increase in surface labeling with phenylephrine. Moreover, each of the changes in p47(phox) targeting mentioned above occurred in dendritic profiles with, or without immunoperoxidase labeling for the AngII AT-1A receptor subtype (AT-1A). These results indicate that chronic administration of agents that induce hypertension can also produce changes in the subcellular localization in p47(phox) in dmNTS neurons. Thus, systemic hypertension may produce alterations in the trafficking of proteins associated with superoxide production in central autonomic neurons, thus revealing a potentially important neurogenic component of free radical production and systemic blood pressure elevation. PMID- 17418123 TI - Neologisms and genericized words in everyday language. PMID- 17418124 TI - Error in antibiotic recommendations. PMID- 17418125 TI - The clinical nurse specialist role. PMID- 17418128 TI - Fragmented publication. PMID- 17418129 TI - Reduce--recycle--reuse: guidelines for promoting perioperative waste management. AB - The perioperative environment generates large amounts of waste, which negatively affects local and global ecosystems. To manage this waste health care facility leaders must focus on identifying correctable issues, work with relevant stakeholders to promote solutions, and adopt systematic procedural changes. Nurses and managers can moderate negative environmental effects by promoting reduction, recycling, and reuse of materials in the perioperative setting. PMID- 17418130 TI - Pediatric medication errors in the postanesthesia care unit: analysis of MEDMARX data. AB - Medication errors involving pediatric patients in the postanesthesia care unit may occur as frequently as one in every 20 medication orders and are more likely to cause harm when compared to medication errors in the overall population. Researchers examined six years of records from the MEDMARX database and used consecutive nonprobability sampling and descriptive statistics to compare medication errors in the pediatric data set to those occurring in the total population data set. Nineteen different causes of error involving 28 different products were identified. The results of the study indicate that an organization can focus on causes of errors and products involved in errors to mitigate future error occurrence. PMID- 17418131 TI - Mentoring high school students in a perioperative setting. AB - Mentoring high school students in perioperative nursing taps an unconventional resource for nursing recruitment. For the past four years, a perioperative nurse has mentored high school students through the Advanced Academic Internship Program of the Miami-Dade County Public School System, Miami, Fla. The students work side by side with their perioperative nurse mentor two or four days a week for the entire school year. Students learn about anatomy related to ophthalmic, orthopedic, and otolaryngology procedures performed at the facility as well as the various roles of health care providers in the perioperative environment. They receive academic grades based on attendance, conduct, and academic performance on a final paper or presentation. PMID- 17418132 TI - Laboratory and diagnostic testing: a perioperative update. AB - Perioperative nurses are challenged to stay informed about the rapidly changing field of diagnostic and laboratory testing. The brain natriuretic peptide test primarily is used to differentiate dyspnea caused by heart failure from dyspnea that results from other causes. Virtual colonoscopy is a promising new alternative for detecting colorectal polyps and cancers. The microalbumin urine test provides an early indication of the development of diabetic complications. This update of relatively new laboratory tests and diagnostic studies also provides implications for perioperative nurses. PMID- 17418133 TI - Quality of life after Roux-en-Y gastric bypass surgery. AB - The incidence of obesity is rising worldwide, leading to a related increase in obesity-associated comorbidities that directly affect longevity and quality of life. Surgical interventions, including the Roux-en-Y gastric bypass procedure, are available for those who have increased risk for morbidity and mortality as a result of repeatedly failed medical management of obesity. Three months after undergoing gastric bypass surgery, patients were sent a survey based on the Impact of Weight on Quality of Life-Lite instrument. The survey results demonstrated marked improvement in overall quality of life and physical function in this population, as well as the ability to decrease or discontinue medications for obesity-related comorbid conditions. PMID- 17418134 TI - Techniques for searching the CINAHL database using the EBSCO interface. AB - The cumulative index to Nursing and Allied Health Literature (CINAHL) is a useful research tool for accessing articles of interest to nurses and health care professionals. More than 2,800 journals are indexed by CINAHL and can be searched easily using assigned subject headings. Detailed instructions about conducting, combining, and saving searches in CINAHL are provided in this article. Establishing an account at EBSCO further allows a nurse to save references and searches and to receive e-mail alerts when new articles on a topic of interest are published. PMID- 17418136 TI - Preparing for electrical power system failures. PMID- 17418138 TI - Versatile retargeting of SH3 domain binding by modification of non-conserved loop residues. AB - Src-homology (SH3) domain belongs to a class of ubiquitous modular protein domains found in nature. SH3 domains have a conserved surface that recognises proline-rich peptides in ligand proteins, but additional contacts also contribute to binding. Using the SH3 domain of hematopoietic cell kinase as a test case, we show that SH3 binding properties can be profoundly altered by modifications within a hexapeptide sequence in the RT-loop region that is not involved in recognition of currently known consensus SH3 target peptides. These results highlight the role of non-conserved regions in SH3 target selection, and introduce a strategy that may be generally feasible for generating artificial SH3 domains with desired ligand binding properties. PMID- 17418139 TI - Crystallographic structure of the SH3 domain of the human c-Yes tyrosine kinase: loop flexibility and amyloid aggregation. AB - SH3 domains from the Src family of tyrosine kinases represent an interesting example of the delicate balance between promiscuity and specificity characteristic of proline-rich ligand recognition by SH3 domains. The development of inhibitors of therapeutic potential requires a good understanding of the molecular determinants of binding affinity and specificity and relies on the availability of high quality structural information. Here, we present the first high-resolution crystal structure of the SH3 domain of the c-Yes oncogen. Comparison with other SH3 domains from the Src family revealed significant deviations in the loop regions. In particular, the n-Src loop, highly flexible and partially disordered, is stabilized in an unusual conformation by the establishment of several intramolecular hydrogen bonds. Additionally, we present here the first report of amyloid aggregation by an SH3 domain from the Src family. PMID- 17418140 TI - The roles of organic anion permeases in aluminium resistance and mineral nutrition. AB - Soluble aluminium (Al(3+)) is the major constraint to plant growth on acid soils. Plants have evolved mechanisms to tolerate Al(3+) and one type of mechanism relies on the efflux of organic anions that protect roots by chelating the Al(3+). Al(3+) resistance genes of several species have now been isolated and found to encode membrane proteins that facilitate organic anion efflux from roots. These proteins belong to the Al(3+)-activated malate transporter (ALMT) and multi-drug and toxin extrusion (MATE) families. We review the roles of these proteins in Al(3+) resistance as well as their roles in other aspects of mineral nutrition. PMID- 17418141 TI - The surface protease PgtE of Salmonella enterica affects complement activity by proteolytically cleaving C3b, C4b and C5. AB - Complement activity in mammalian serum is fundamentally based on three homologous components C3b, C4b and C5. During systemic infection, the gastrointestinal pathogen Salmonella enterica disseminates within host phagocytic cells but also extracellularly. Consequently, systemic Salmonella transiently confronts the complement system. We show here that the surface protease PgtE of S. enterica proteolytically cleaves C3b, C4b and C5 and that the expression of PgtE enhances bacterial resistance to human serum. Degradation of C3b was further enhanced by PgtE-mediated plasminogen activation. PMID- 17418142 TI - K+ channel activity in plants: genes, regulations and functions. AB - Potassium (K(+)) is the most abundant cation in the cytosol, and plant growth requires that large amounts of K(+) are transported from the soil to the growing organs. K(+) uptake and fluxes within the plant are mediated by several families of transporters and channels. Here, we describe the different families of K(+) selective channels that have been identified in plants, the so-called Shaker, TPK and Kir-like channels, and what is known so far on their regulations and physiological functions in the plant. PMID- 17418143 TI - Extracellular lipid droplets promote hemozoin crystallization in the gut of the blood fluke Schistosoma mansoni. AB - Hemozoin (Hz) is a heme crystal produced upon hemoglobin digestion as the main mechanism of heme disposal in several hematophagous organisms. Here, we show that, in the helminth Schistosoma mansoni, Hz formation occurs in extracellular lipid droplets (LDs). Transmission electron microscopy of adult worms revealed the presence of numerous electron-lucent round structures similar to LDs in gut lumen, where multicrystalline Hz assemblies were found associated to their surfaces. Female regurgitates promoted Hz formation in vitro in reactions partially inhibited by boiling. Fractionation of regurgitates showed that Hz crystallization activity was essentially concentrated on lower density fractions, which have small amounts of pre-formed Hz crystals, suggesting that hydrophilic hydrophobic interfaces, and not Hz itself, play a key catalytic role in Hz formation in S. mansoni. Thus, these data demonstrate that LDs present in the gut lumen of S. mansoni support Hz formation possibly by allowing association of heme to the lipid-water interface of these structures. PMID- 17418144 TI - Activation of intracellular signaling pathways is necessary for an increase in VDR expression and its nuclear translocation. AB - 1,25-Dihydroxyvitamin D(3) (1,25D) regulates gene transcription through the nuclear vitamin D receptor (VDR) and initiates rapid cellular responses via an unknown mechanism. Here we report that 1,25D induces a rapid increase in synthesis of VDR protein and its transport to the nucleus. These results are similarly obtained in myeloid leukemia cell lines, and in blast cells from blood of patients diagnosed with acute myeloid leukemia, subtypes M2 and M4. Our results suggest that stability of unliganded VDR is LY294002- and PD98059 dependent, and that ligation of VDR leads to its increased translation and nuclear translocation. The receptor localized in the cell nucleus is not exported back to the cytosol by exportin 1. We also show that the cytosolic portion of VDR in leukemia cells is localized in the vicinity of the plasma membrane, close to the F-actin cytoskeleton. PMID- 17418145 TI - T0901317 is a potent PXR ligand: implications for the biology ascribed to LXR. AB - The liver X receptors (LXRalpha and beta) are nuclear receptors that coordinate carbohydrate and lipid metabolism. Insight into the physiologic roles of the LXRs has been greatly facilitated by the discovery of potent synthetic agonists. Here we show that one of these compounds, T0901317, is also a high-affinity ligand for the xenobiotic receptor pregnane X receptor (PXR). T0901317 binds and activates PXR with the same nanomolar potency with which it stimulates LXR activity. T0901317 induces expression not only of LXR target genes, but also of PXR target genes in cells and animals, including the scavenger receptor CD36, a property not shared by more specific LXR ligands, such as GW3965. Activation of PXR targets may explain why T0901317 induces dramatic liver steatosis, while GW3965 has a milder effect. These results suggest that many of the biological activities heretofore associated with LXR activation may be mediated by PXR, not LXR. Since T0901317 has been widely used in animals to study LXR function, the in vivo effects of this compound ascribed to LXR activation should be re-examined. PMID- 17418147 TI - Letter from the editors. PMID- 17418146 TI - Spatial regulation of exocytosis and cell polarity: yeast as a model for animal cells. AB - Exocytosis is the major mechanism by which new membrane components are delivered to the cell surface. In most, if not all, eukaryotic cells this is also a highly spatially regulated process that is tightly coordinated with the overall polarity of a cell. The Rho/Cdc42 family of GTPases and the lethal giant larvae/Sro7 family are two highly conserved families of proteins which appear to have dual functions both in cell polarity and exocytosis. Analysis of their functions has begun to unravel the coordination between these processes and propose a model for polarized vesicle docking and fusion at the site of asymmetric cell growth. PMID- 17418148 TI - Age-related structural and functional changes in the breast: multimodality correlation with digital mammography, computed tomography, magnetic resonance imaging, and positron emission tomography. AB - Preliminary results generated from digital mammography, computed tomography, magnetic resonance imaging, and (18)F-fluorodeoxyglucose positron emission tomography demonstrate concordant findings of decreasing glandular tissue and decreasing metabolic activity with increasing age. These results are presented in the context of a detailed literature review summarizing age-related changes in the breast, both from the histologic/physiologic and the imaging perspectives. We also discuss potential applications of this approach and emphasize the importance of new advanced imaging technologies to offer high levels of quantitative precision for tissue characterization for research and clinical purposes. PMID- 17418149 TI - Assessment of age-related changes in abdominal organ structure and function with computed tomography and positron emission tomography. AB - With the size of the aged population in the United States expected to grow considerably during the next several decades, the number of imaging studies performed on such aged individuals will similarly increase. Thus, it is important to understand normal age-related changes in the structural and functional imaging appearance of the abdominal organs. We therefore present preliminary data and a review of the literature relevant to structural and functional changes in the abdominal organs of children and older adults. In a retrospective study of both adult and pediatric populations, we used computed tomography (CT), positron emission tomography (PET), and PET/CT imaging to investigate age-associated changes in size, attenuation, and metabolic function of the abdominal organs. Organs of interest include the liver, spleen, pancreas, kidneys, adrenal glands, stomach, small bowel, colon, and rectum. Although volumes of adult liver, spleen, pancreas, and kidneys do not change significantly with age, adult left and right adrenal gland volumes do significantly increase with age (r = 0.2823, P = 0.0334, and r = 0.3676, P = 0.0049, respectively). Also, the attenuation of adult liver (r = -0.2122, P = 0.0412), spleen (r = -0.4508, P < 0.0001), pancreas (r = 0.5124, P = 0.0007), and left and right adrenal gland (r = -0.5835, P < 0.0001 and r = -0.6135, P < 0.0001, respectively) decrease significantly with increasing age. Every organ studied in the pediatric population demonstrates a positive association between organ volume and age. Significant age-related changes in organ function are noted in the adult liver and small bowel, with the liver demonstrating a positive association between metabolic activity and age (r = 0.4434, P = 0.0029) and the small bowel showing an inverse association between mean small bowel standardize uptake value and age (r = -0.2435, P = 0.0174). Also, the maximum overall small bowel and colon metabolic activity in children increases with age (r = 0.6478, P = 0.0008). None of the other organs studied (ie, spleen, pancreas, adrenal glands, stomach, colon, rectum) demonstrate significant changes in metabolism with advancing age. The metabolic volumetric product (calculated as the product of organ volume and mean organ SUV) of the liver and spleen does not change significantly with age. In conclusion, various abdominal organs demonstrate differential changes in volume, attenuation, and/or metabolism with increasing age in pediatric and adult populations. PMID- 17418150 TI - Age-related structural and metabolic changes in the pelvic reproductive end organs. AB - In this work, we provide preliminary data and a review of the literature regarding normal structural and functional changes that occur in the aging uterus, ovary, testicle, and prostate gland. It is expected that such knowledge will help physicians to distinguish physiologic changes from pathologic changes at an early stage. We retrospectively reviewed pelvic magnetic resonance imaging (MRI) scans of 131 female and 79 male subjects ages 13 to 86 years to determine changes in volume of the uterus, ovary, and prostate gland with age. Scrotal ultrasound examinations of 150 male subjects ages 0 to 96 years also were analyzed retrospectively to determine changes in testicular volume with age. In addition, (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG-PET) scans of 145 male subjects ages 11 to 90 years were analyzed retrospectively to assess for changes in maximum standardized uptake value (SUV(max)) of the testicles with age. The uterus had a mean volume of 38.55 +/- 3.68 cm(3) at 17 to 19 years of age, increased to a peak volume of 71.76 +/- 19.81 cm(3) between 35 to 40 years, and then declined to 24.02 +/- 8.11 cm(3) by the eighth decade of life. The maximal ovarian volume per subject maintained a relatively stable size in early life, measuring 9.46 +/- 3.25 cm(3) during the second decade of life, 8.46 +/- 3.32 cm(3) in the mid-fourth decade of life, and 7.46 +/- 3.33 cm(3) at 45 years of age, after which it declined to 4.44 +/- 2.02 cm(3) by the late fifth decade of life. The ovaries were not identifiable on MRI in subjects beyond the sixth decade of life. The volume of the prostate increased from 23.45 +/- 6.20 cm(3) during the second decade of life to 47.5 +/- 41.59 cm(3) by the late eighth decade of life; the central gland of the prostate increased from 9.96 +/- 3.99 cm(3) to 29.49 +/- 28.88 cm(3) during the same age range. Mean testicular volume was 11.2 +/- 5.9 cm(3). Testicular volume increased with age from birth to 25 years. After age 25, there was a significant decline in the testicular volume. The mean SUV(max) for the testicles was 1.9 +/- 0.5. Testicular metabolic activity demonstrated an increasing trend until the age of 35 years. A plateau in SUV(max) was observed after the age of 35 years until the age of 65 years. A slight decrease in SUV(max) was observed after the age of 65 years. The pelvic structures of men and women change both structurally and functionally over the lifespan, and such changes can be quantified using ultrasound, MRI, and (18)F-FDG PET. PMID- 17418151 TI - Structural and functional imaging of normal bone marrow and evaluation of its age related changes. AB - A number of noninvasive imaging techniques have been used for the evaluation of bone marrow, including magnetic resonance imaging (MRI) and bone marrow scintigraphy. The appearance of bone marrow on MRI varies considerably depending on the proportion of red and yellow marrow, and the composition of the red marrow and its distribution with relation to age and sex. The composition of bone marrow also can vary under physiological and pathological conditions. MRI is a highly sensitive technique for evaluating the bone marrow, but it is limited in its practical use for whole-body bone marrow screening. Bone marrow scintigraphy with radiolabeled compounds such as technetium-99m-labeled nanocolloid and monoclonal antibodies has the advantage of evaluating the entire bone marrow, and has been used for the diagnosis of various bone marrow disorders. In addition, (18)F fluorodeoxyglucose positron emission tomography (FDG-PET) imaging can be used to evaluate bone marrow metabolism and disease and to provide information about the state of the primary tumor, lymph nodes, and distant metastases. Understanding of the appearance of normal bone marrow, including age- and sex-specific differences with each of these imaging modalities, is essential to permit accurate diagnosis of benign and malignant bone marrow disorders. We present a review of MRI and scintigraphy of normal bone marrow with some emphasis on FDG-PET imaging in assessing marrow activity in normal and abnormal states and also present preliminary data regarding normal age-related changes in bone marrow through use of FDG-PET, as well as the role of segmentation of bone marrow on MRI for quantitative calculation of the metabolic volumetric product for red marrow metabolism using FDG-PET. PMID- 17418152 TI - Determination of age-related changes in structure and function of skin, adipose tissue, and skeletal muscle with computed tomography, magnetic resonance imaging, and positron emission tomography. AB - In this article, we report quantitative preliminary data obtained from retrospective analysis of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) and combined PET-computed tomography (PET/CT) examinations in subjects ages 3 to 84 years pertaining to changes in the metabolism of skin, subcutaneous adipose tissue, visceral adipose tissue, and skeletal muscle with age, as well as age-related changes in skeletal muscle attenuation. We also propose a new method for identifying hypermetabolic brown fat on FDG-PET. Finally, we present a review of the literature regarding reported age-related structural and functional changes that occur in skin, fat, and skeletal muscle. Using FDG-PET, We evaluated 213 subjects for changes in the metabolism of skin, adipose tissue, and skeletal muscle with aging. Thirty-two separate subjects were chosen to measure maximum standardized uptake value (SUV) of hypermetabolic brown fat on dual-time point PET imaging. Finally, 15 subjects evaluated by PET/CT were selected to measure changes in metabolism and attenuation of skeletal muscle, and changes in metabolism of adipose tissue with aging. We found that skin, fat, and skeletal muscle all demonstrate significant (P < 0.05) increases in SUV with increasing age on PET imaging. Dual-time point PET imaging demonstrates increasing FDG uptake of hypermetabolic brown fat in various regions studied. Finally, our PET/CT studies revealed statistically insignificant (P > 0.05) decreases in SUV of adipose tissue with aging and the opposite trend in skeletal muscles (P > 0.05). Skeletal muscle attenuation in the various regions studied was found to significantly decrease with age (P < 0.05). Our study shows notable trends in metabolism and attenuation of skeletal muscle and metabolism of skin and adipose tissue that occur with normal aging. We hope that the methodologies and data we present here will serve as a useful starting point for those interested in conducting future prospective research on age-related changes in these structures. PMID- 17418153 TI - Increased (18)F-fluorodeoxyglucose uptake in benign, nonphysiologic lesions found on whole-body positron emission tomography/computed tomography (PET/CT): accumulated data from four years of experience with PET/CT. AB - The use of (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG-PET) in the field of oncology is rapidly evolving; however, (18)F-FDG is not tumor specific. Aside from physiological uptake (18)F-FDG also may accumulate in benign processes. Knowledge of these (18)F-FDG-avid nonmalignant lesions is essential for accurate PET interpretation in oncologic patients to avoid a false-positive interpretation. Through the systematic review of the reports of PET/computed tomography (CT) studies performed in oncologic patients during a 6-month period, we found benign nonphysiological uptake of (18)F-FDG in more than 25% of studies. In half of these, (18)F-FDG uptake was moderate or marked in intensity, similar to that of malignant sites. A total of 73% of benign lesions were inflammatory in nature, with post-traumatic bone and soft-tissue abnormalities (including iatrogenic injury) and benign tumors accounting for the remainder. The differentiation of benign from malignant uptake of (18)F-FDG on PET alone may be particularly challenging as a result of the low anatomical resolution of PET and paucity of anatomical landmarks. Fusion imaging, namely PET/CT, has been shown to improve not only the sensitivity of PET interpretation but also its specificity. Aside from better anatomical localization of lesions on PET/CT, morphological characterization of lesions on CT often may improve the diagnostic accuracy of nonspecific (18)F-FDG uptake. Correlation with CT on fused PET/CT data may obviate the need for further evaluation or biopsy in more than one-third of scintigraphic equivocal lesions. Familiarity with (18)F-FDG-avid nonmalignant lesions also may extend the use of (18)F-FDG-PET imaging beyond the field of oncology. We have tabulated our experience with benign entities associated with increased (18)F-FDG uptake on whole-body PET/CT from 12,000 whole-body (18)F-FDG PET/CT studies performed during a 4-year period. PMID- 17418154 TI - Novel quantitative techniques for assessing regional and global function and structure based on modern imaging modalities: implications for normal variation, aging and diseased states. AB - In this review, we describe the current approaches used for quantitative assessment of regional and global function with positron emission tomography (PET) imaging (combined with structural imaging modalities) with emphasis on both research and clinical applications of this powerful approach. We particularly refer to the impact of such measurements in assessing physiological processes such as aging and measuring response to treatment in serious disorders such as cancer. Although a multitude of methods has been described in literature, the optimal approaches that are both accurate and practical in clinical settings need to be defined and refined. Standardized uptake value (SUV) continues to be the most widely used index in the current practice. Calculating SUV at a single time point and assigning standard regions of interest are inadequate and suboptimal for the purposes adopted by the medical community. The concepts of partial volume correction for measured values in small lesions, dual-time point and delayed PET imaging, and global metabolic activity for assessment of various stages of disease may overcome deficiencies that are associated with the current quantitative (ie, SUV) techniques. Serious consideration of these concepts will enhance the role and reliability of these quantitative techniques, and therefore compliment the World Health Organization or the Response Evaluation Criteria in Solid Tumors (RECIST) criteria for managing patients with cancer and other disorders, including physiological states such as aging and serious diseases such as atherosclerosis and neurological diseases. We also introduce the concepts that allow for segmentation of various structural components of organs like the brain for accurate measurement of functional parameters. We also describe complicated kinetic modeling and methodologies that have been used frequently for assessing metabolic and pharmacological parameters in the brain and other organs. Simplified quantitative techniques based on these concepts are described, but should be validated against the kinetic models to test their role as practical tools. PMID- 17418155 TI - Biologic markers of ovarian reserve and reproductive aging: application in a cohort study of HIV infection in women. AB - OBJECTIVE: To compare Mullerian inhibiting substance (MIS) levels in serum obtained during the early follicular phase to those obtained randomly during the menstrual cycle. To determine whether HIV infection influences early follicular MIS levels, an early marker of ovarian aging. DESIGN: A cross-sectional study. SETTING: Women's Interagency HIV Study, a multicenter prospective study. PATIENT(S): Serum samples obtained from 263 (187 HIV infected and 76 uninfected) participants of the Women's Interagency HIV Study who reported menstrual bleeding during the preceding 6 months and who were not taking exogenous hormones. INTERVENTION(S): Early follicular (cycle days 2-5) MIS samples were compared with serum samples that had been obtained without regard to menstrual cycle phase. Comparison samples were obtained within 6 weeks before or within 3 to 6 months after the early follicular samples. Early follicular FSH, E(2), inhibin B, and MIS levels were also compared between the HIV infected and uninfected women. MAIN OUTCOME MEASURE(S): Correlation between early follicular MIS and prior and subsequent samples. Comparison of serum markers of ovarian reserve between HIV positive and negative women. RESULT(S): The MIS values from early follicular and other random cycle phases were highly correlated with each other (r > 0.93). In multivariate analysis, increased age and FSH level and lower inhibin B levels were associated with lower MIS level; MIS values did not vary by HIV serostatus. CONCLUSION(S): Without regard to cycle phase, MIS was similar during early follicular phase and highly correlated with early follicular FSH and inhibin B in women with and without HIV. Measurement of serum MIS offers a simplified method of determining ovarian reserve using specimens obtained without menstrual phase timing. Furthermore, using biologic measures of reproductive aging, we found no evidence that HIV infection influences ovarian aging. PMID- 17418156 TI - Expression and cyclic variations of catechol-O-methyl transferase in human endometrial stroma. AB - OBJECTIVE: To investigate the role of catechol-O-methyl transferase (COMT) in the regulation of estrogen metabolism in human endometrium. DESIGN: Laboratory study. SETTING: Academic research laboratory. INTERVENTION(S): Immunohistochemistry was used to localize COMT protein in human endometrial tissues. Catechol-O-methyl transferase promoter-luciferace reporter gene transactivation assay was used to assess COMT promoter activity in response to estrogen and progesterone treatment in primary human endometrial stroma (pHES) cells. Catechol-O-methyl transferase protein and mRNA expression were determined by Western blot and/or real-time polymerase chain reaction. The effect of 2-methoxy estrogen treatment on DNA proliferation, B-cell lymphoma 2, and vascular epithelial growth factor protein expression were assessed by Hoechst and Western blot analyses, respectively. MAIN OUTCOME MEASURE(S): Catechol-O-methyl transferase protein and mRNA subcellular localization and expression in human endometrial tissues and pHES cells. RESULT(S): Catechol-O-methyl transferase protein expression in human endometrial tissues was up-regulated in the proliferative phase and down-regulated in the midsecretory phase of the menstrual cycle. Estrogen induced a dose-dependent increase in COMT proximal promotor-luciferace transactivation in pHES cells whereas progesterone inhibited it. Estrogen up-regulated soluble COMT protein isoform expression whereas the addition of progesterone down-regulated it in pHES cells. High doses of 2-methoxy estrogen inhibited endometrial stroma cell proliferation, and down-regulated B-cell lymphoma 2 and vascular epithelial growth factor protein expression. CONCLUSION(S): Catechol-O-methyl transferase expression is hormonally regulated in human endometrial stroma. Catechol-O-methyl transferase product, 2-methoxy estrogen, inhibited endometrial stroma cell proliferation and decreased vascular epithelial growth factor and B-cell lymphoma 2 protein expression. PMID- 17418157 TI - Mutation analysis of NANOS3 in 80 Chinese and 88 Caucasian women with premature ovarian failure. AB - NANOS3 encodes an RNA-binding protein and has a conserved function in germ cell development. Our objective was to investigate whether mutations in NANOS3 were present in Chinese and Caucasian women with premature ovarian failure. A known synonymous single-nucleotide polymorphism (rs 2016163) in exon 1 was identified through sequencing 80 Chinese and 88 Caucasian women with premature ovarian failure. No additional single-nucleotide polymorphisms or mutations were found in exons encoding for NANOS3. Our findings suggest that mutations in NANOS3 exons are rare in both Chinese and Caucasian women with premature ovarian failure. PMID- 17418158 TI - Social concerns of women undergoing infertility treatment. AB - OBJECTIVE: Our study was undertaken to determine [1] what women are disclosing to their employer with regard to their infertility, [2] what demographic characteristics are associated with women who are more likely to disclose, and [3] if there is an association between disclosure and lowering one's stress. We hypothesize that, in certain women, disclosure may lower stress, and therefore increase success rate of in vitro fertilization. DESIGN: Cross-sectional questionnaire. SETTING: University Infertility Treatment Center. PATIENT(S): We handed out a questionnaire to patients being evaluated and treated for infertility over a 6-month period. A total of 267 questionnaires were handed out and all were collected. MAIN OUTCOME MEASURE(S): We collected demographic data as well as information regarding privacy, disclosure, and stress. We then compared women who disclose to their employer that they are being seen by an infertility specialist to those women who do not disclose. We also measured stress and determined if higher stress level was associated with disclosure or nondisclosure. RESULT(S): Most women who did disclose did so because they needed a reason to leave work for frequent doctor visits. Among women who did not disclose, the main reason for nondisclosure was to protect their privacy. Women with a high school education were more likely to disclose compared with those with a college and postgraduate education. African American/Caribbean American women were least likely to disclose. Those who were out of work more often because of their infertility were more likely to disclose. There was not an association with disclosure and decreasing stress level. CONCLUSION(S): Women who did or did not disclose their infertility status to their employer were different with regard to level of education, race/ethnicity, and number of visits per month to the doctor. The decision to disclose does not seem to have a significant impact on stress. PMID- 17418159 TI - Severity of energy-related menstrual disturbances increases in proportion to indices of energy conservation in exercising women. AB - Alterations in resting energy expenditure and metabolic hormones (energy conservation) are evident in increasing magnitude across a continuum of increasing severity of clinical menstrual disturbances, including luteal-phase defects, anovulation, and amenorrhea in exercising women. These data provide further evidence of the tight association between energy balance and reproduction and suggest that subtle declines in energy availability can produce clinically recognized menstrual disturbances. PMID- 17418160 TI - 5-ASA and colorectal cell-cycle progression. PMID- 17418162 TI - IBD legislation introduced. PMID- 17418163 TI - Role of impedance/pH monitoring in refractory gerd: let's be careful out there! PMID- 17418164 TI - Got guts? Need nerve! PMID- 17418167 TI - HCV clearance among hemophiliacs and beta-thalassemics. PMID- 17418168 TI - It's chickens and eggs all over again: is central reorganization the result or cause of persistent visceral pain? PMID- 17418169 TI - Vaccine-induced T-cell responses against HCV: one step taken, more to follow. PMID- 17418171 TI - AGA position statement of computed tomographic colonography. PMID- 17418172 TI - Clinical Challenges and Images in GI. Microvillous inclusion disease. PMID- 17418174 TI - Treatment of hepatitis C: don't put all your eggs in one basket! PMID- 17418175 TI - Are all proton pump inhibitors created equal? PMID- 17418176 TI - Solubilisation and binding characteristics of a recombinant beta2-adrenergic receptor expressed in the membrane of Escherichia coli for the multianalyte detection of beta-agonists and antagonists residues in food-producing animals. AB - The number of substances with beta-agonistic activity, illegally introduced in meat production or in sports doping as anabolic or beta-blocking agents is increasing. Analytical methods suited for their multianalyte detection are thus necessary. In this perspective, receptor assays were developed. The research activities undertaken in this study describe the solubilisation of a recombinant human beta(2)-adrenergic receptor produced in the inner membrane of genetically modified Escherichia coli, using the detergent n-dodecyl-beta-d-maltoside. Its potential to detect the presence of beta-agonists or beta-blockers in biological samples was evaluated. The solubilised beta(2)-adrenergic receptor retained its binding affinity in a radio-receptor assay based on the competition for the binding to receptors between a ligand (beta-agonist or antagonist) and the radioligand [(125)I]iodocyanopindolol. The IC(50) values ranged from 5+/-1 x 10( 8) M (clenbuterol) to 8+/-2 x 10(-6) M (isoxsuprine) for the beta-agonists tested and from 1.5+/-0.2 x 10(-10) M (carazolol) to 1.2+/-0.2 x 10(-5) M (metoprolol) for the beta-blockers tested. It was shown to have a lower limit of detection than a radio-receptor assay using the solubilised beta(2)-adrenoceptor expressed in a mammalian cell line. The solubilised recombinant human beta(2) adrenoreceptor expressed in E. coli would be a useful tool to develop non radioactive multianalyte screening methods. PMID- 17418177 TI - Highly sensitive phage-based biosensor for the detection of beta-galactosidase. AB - Development of real-time sensor based on the target-specific probe that make possible sensitive, rapid and selective detection and monitoring of the particular antigen molecules could be of substantial importance to the many applications. Because of its high specificity to the target molecules, excellent temperature stability, and easy production, bacterial phage might serve as a powerful biorecognition probe in biosensor applications. Here, we report extremely sensitive and specific label-free direct detection of model antigen, beta-galactosidase (beta-gal), based on surface plasmon resonance (SPR) spectroscopy. The beta-gal specific landscape phage 1G40 has been immobilized on the gold surface of SPR SPREETA sensor chip through physical adsorption [V. Nanduri, A.M. Samoylova, V.Petrenko, V. Vodyanoy and A.L.Simonian, Comparison of optical and acoustic wave phage biosensors, 206th Meeting of The Electrochemical Society, Honolulu, Hawaii, October 3-8, (2004)]. Another non-specific to the beta gal phage, a wild-type phage F8-5, was used in the reference channel. The concentration-dependent binding of beta-gal in both channels were assessed by monitoring the sensor optical response as a function of time under different experimental conditions, and the concentration of beta-gal was computed in differential mode. Concentrations of beta-gal between 10(-12) M and 10(-7) M could be readily detected, with linear part of calibration curve between 10(-9) M and 10(-6) M. When beta-gal was pre-incubated with different concentrations of free 1G40 phage prior to exposure to the biosensor, concentration-dependent inhibition was observed, indicating on biosensor high specificity toward beta gal. Apart from a flow through mode used to deliver the samples to the surface for the SPR sensor, batch mode sensing was also employed to study the binding of beta-gal to immobilized phage on the SPR sensor surface. Experiments using a flow through mode provided more consistent results in the full dose range and showed higher sensitivity as opposed to the batch mode studies. The mean K(d) and binding valences for the flow through mode studies was 1.3+/-0.001 nM and 1.5+/ 0.03, in comparison to 26+/-0.003 nM and 2.4+/-0.01 for the batch mode studies. The average thickness of phage 1G40 adlayer deposited through flow through and batch mode was 3+/-0.002 and 0.66+/-0.001 nm, respectively. PMID- 17418178 TI - Multiple enzyme linked immunosorbent assay system on a capillary-assembled microchip integrating valving and immuno-reaction functions. AB - Multiple enzyme linked immunosorbent assay (ELISA) chip is developed by using capillary-assembled microchip (CAs-CHIP) technique, which involves simple embedding of 2-3mm length of square capillaries possessing valving and immuno reaction functions into the microchannels fabricated on a PDMS substrate. In contrast to the previously reported ELISA chips, our system enables not only the flexible design of the multi-ELISA chip required for many different diagnostic purposes, but also the valving operation required for a reliable analysis. Here, a thermo-responsive polymer-immobilized capillary was used together with a small Peltier device, as a valving part, and different antibody-immobilized capillaries were used as immuno-reaction part. Sample solution and detecting reagent solutions were sequentially introduced through the valving capillary, and the valve is closed to completely stop the solution flow inside the immuno-reaction capillaries and detected using thermal lens microscope (TLM). Different anti-IgGs (human, goat, chicken) were immobilized and used as ELISA parts of CAs-CHIP. Sequential introductions of the mixed IgG solution, mixed enzyme-antibody solution and substrate solution facilitated the multiple determinations of 0.1 ng mL(-1) IgGs (human, goat, chicken) with total analysis time of about 30 min. The valve-integrated multi-ELISA chip developed here can be applied for many different diagnostic purposes by using different immuno-reaction capillaries necessary for a specific clinical diagnostic application. PMID- 17418179 TI - Selective separation of hydroxy polychlorinated biphenyls (HO-PCBs) by the structural recognition on the molecularly imprinted polymers: direct separation of the thyroid hormone active analogues from mixtures. AB - We developed novel separation media for hydroxy polychlorinated biphenyls (HO PCBs) using the molecular imprinting techniques. The results of evaluation for the molecularly imprinted polymers (MIPs) by the liquid chromatography (LC) suggested that MIPs had selective separation ability for certain HO-PCB analogues. The results of the LC evaluations and molecular modeling indicated that the molecular volumes and pK(a) values of template molecules were related with the retention factor of HO-PCBs. Additionally, according to the detail evaluation toward the selective separation behaviors of MIPs, these HO-PCB analogues have low pK(a) values dependent on their chemical structures. In other words, the prepared MIPs had selective recognition ability against the analogues, which have an OH group on a phenyl carbon and two chlorine atoms on the both neighboring carbons of the carbon attached with the OH group. Moreover, these analogues may have a potential for thyroid hormone activities so that we attempted to separate these analogues directly from mixtures of HO-PCBs using a prepared MIP. PMID- 17418180 TI - Use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide for rapid detection of methicillin-resistant Staphylococcus aureus by resonance light scattering. AB - A rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) using resonance light scattering (RLS) on an ordinary fluorescence spectrometer was developed. The viable MRSA reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to produce insoluble particles which displayed intense resonance scattering light. It showed a linear relationship between the number of viable MRSA and the RLS intensity. Dead MRSA were unable to reduce MTT. MRSA exposed to flavonoids extracted from Marchantia convoluta (MCF) showed a MCF concentration-dependent inhibition of the ability to reduce MTT. The RLS could, in combination with the MTT assay, be a rapid and sensitive detection method for vitro-cultured MRSA. PMID- 17418181 TI - Raman spectroscopic method for the determination of medroxyprogesterone acetate in a pharmaceutical suspension: validation of quantifying abilities, uncertainty assessment and comparison with the high performance liquid chromatography reference method. AB - An alternative fast and non-destructive validated Raman spectroscopic analytical procedure, requiring no sample preparation, was compared with the industrially applied HPLC reference method (Pfizer Manufacturing Belgium) for the quantitative determination of medroxyprogesterone acetate (MPA) in DepoProvera suspensions (150 mg mL(-1), Pfizer). The Raman calibration model was developed by plotting the peak intensity of the baseline-corrected and normalized spectral band (corrected by external standard measurements) between 1595 and 1620 cm(-1) against known MPA concentrations in standards. At this band, no spectral interferences from the suspension medium are observed. The most suitable model for the calibration data (straight line or higher order polynomial) was determined by evaluating the fit and predictive properties of the models. In a second step, the developed Raman spectroscopic analytical method was validated by calculating the accuracy profile on the basis of the analysis results of validation samples. Furthermore, based on the data of the accuracy profile, the measurement uncertainty was determined. Finally, as the aim of the alternative method is to replace the destructive, time-consuming HPLC method, requiring sample preparation, it needs to be demonstrated that the new Raman method performs at least as good as the HPLC method. Therefore, the performance (precision and bias) of both methods was compared. A second order polynomial calibration curve through the calibration data supplies the best predictive properties and gives an acceptable fit. From the accuracy profile, it was concluded that at the target concentration (150 mg mL(-1)), 95 out 100 future routine measurements will be included within the acceptance limits (5%). Comparison of the alternative method with the reference method at the target concentration indicates that the Raman method performs at least as good as the HPLC method for precision (repeatability and intermediate precision) and bias. The fast and non-destructive Raman method hence provides an alternative for the destructive and time-consuming HPLC procedure. PMID- 17418182 TI - A new method for testing synthetic drugs adulterated in herbal medicines based on infrared spectroscopy. AB - Developing a simple and fast method to analyze possibly adulterated synthetic drugs in suspected herbal medicines (HM) is both methodologically and commercially significant. This paper constructs a new approach named local straight-line screening (LSLS), to the solution of the problem, after carefully observing the characteristics of the spectral line shapes. LSLS can be applied to both the qualitative and quantitative analysis of suspected HM, based solely on infrared spectrum of the possible synthetic drug and the suspected HM without any sample pretreatment. The concept and the application of the method are exemplified by analysis of sibutramine hydrochloride (SH), an anti-obesity medicine, adulterated in some commercially available HM-based diet pills, and compared with MSFDSS, and SNICA methods. The results show that, despite highly overlapping spectra and unresolved data, it is possible with LSLS to obtain an accurate discrimination and determination of SH in the HM-based diet pills. This allows the method to be potentially used in the primary screening of other synthetic drugs suspiciously adulterated in HM, with high rapidity, accuracy and cost-effectiveness. PMID- 17418183 TI - Use of linear discriminant analysis applied to vibrational spectroscopy data to characterize commercial varnishes employed for art purposes. AB - An improvement of methodologies for characterising synthetic resins used in varnishes employed for art purposes has been suggested. Several kinds of standard of the most common polymeric resins (acrylic, vinyl, poly(vinyl alcohol), alkyd, cellulose nitrate, latex, polyester, polyurethane, epoxy, organosilicic, and ketonic) were analyzed by Fourier transform infrared (FTIR) spectroscopy. Synthetic resins characterization is based on the mathematical treatment of their whole spectrum, dividing it in 13 sections, avoiding the one-by-one interpretation of the absorption bands. The mathematical model takes as variables the maximal absorbance of each section, and each synthetic standard resin as categories. Two exploratory analysis methods, Hierarchical Clustering and Principal Component Analysis (PCA), and a classificatory chemometric tool, linear discriminant analysis (LDA), are tested, validating the models by leave-one-out method. LDA is proved to be a powerful tool for grouping objects in categories, providing a satisfactory distinction of polymeric resin standards. The described analytical procedure has successfully been applied to characterization of synthetic resins contained in commercial varnishes. PMID- 17418184 TI - Independent component analysis as a pretreatment method for parallel factor analysis to eliminate artefacts from multiway data. AB - Parallel factor analysis (PARAFAC) has successfully been used in many applications for the analysis of excitation-emission fluorescence data. However, some measurement "artefacts", such as Rayleigh or Raman scattering, can pose a problem for the extraction of the PARAFAC components and their interpretation. Replacing the spectral zones corresponding to these signals by missing values in the data is not necessarily a method of choice in the cases where informative signals lie in the same wavelength regions. In this article, independent component analysis (ICA) is used on the unfolded cubic array, and the independent components related to the Rayleigh and Raman scattering are identified and removed prior to the reconstruction of the excitation-emission fluorescence data cube. PARAFAC is then applied on these data reconstructed after selective artefact removal, and satisfactory models can be obtained. This procedure, although particularly useful for 3D fluorescence data, may be applied to other types of data as well. PMID- 17418185 TI - Headspace liquid-phase microextraction of methamphetamine and amphetamine in urine by an aqueous drop. AB - This study developed a headspace liquid-phase microextraction (LPME) method by using a single aqueous drop in combination with high performance liquid chromatography (HPLC)-UV detection for the determination of methamphetamine (MAP) and amphetamine (AP) in urine samples. The analytes, volatile and basic, were released from sample matrix into the headspace first, and then protonated and dissolved in an aqueous H(3)PO(4) drop hanging in the headspace by a HPLC syringe. After extraction, this drop was directly injected into HPLC. Parameters affecting extraction efficiency were investigated and optimized. This method showed good linearity in the investigated concentration range of 1.0-1500 microg L(-1), repeatability of the extraction (R.S.D.<5%, n=6), and low detection limits (0.3 microg L(-1) for both analytes). Enrichment factors of about 400-fold and 220-fold were achieved for MAP and AP, respectively, at optimum conditions. The feasibility of the method was demonstrated by analyzing human urine samples. PMID- 17418186 TI - Coupling continuous ultrasound-assisted extraction with ultrasonic probe, solid phase extraction and high-performance liquid chromatography for the determination of sodium Danshensu and four tanshinones in Salvia miltiorrhiza bunge. AB - A dynamic continuous ultrasound-assisted extraction with high intensity ultrasonic probe (CUAE-HIUP) combined with solid-phase extraction (SPE) for preconcentration and clean-up of the extract prior to high-performance liquid chromatography (HPLC) determination of the main biological active ingredients, sodium Danshensu and four tanshinones (dihydrotanshione I, tanshinone I, cryptotanshinone and tanshinone IIA) from root of Salvia miltiorrhiza bunge has been developed. An experimental design (Plackett-Burman design, 2(6)x3/16) was used to optimize the CUAE-HIUP procedure and the SPE step. Detection limits of 1.2-4.2 x 10(-6) mg mL(-1) were achieved with the preconcentration efficiency of more than 10-folds by the SPE procedure. The repeatability and within-laboratory reproducibility of the whole process, expressed as relative standard deviations (RSDs), were 1.5-3.6%, 1.6-3.1%, respectively. As compared with the conventional extraction techniques such as room temperature extraction (24 h), Soxhlet (4 h) and even microwave-assisted extraction (2 min), CUAE-HIUP achieved the highest extraction yield (98.9%) and the least amount (10 mL) of solvent compared with the other techniques (16 mL) in only 3 min. The method was successfully applied to the determination of the five biological active ingredients in root of S. miltiorrhiza bunge and Danshen-containing pharmaceutical formulations. PMID- 17418187 TI - The determination of organochlorine pesticides based on dynamic microwave assisted extraction coupled with on-line solid-phase extraction of high performance liquid chromatography. AB - A rapid technique based on dynamic microwave-assisted extraction coupled with on line solid-phase extraction of high-performance liquid chromatography (DMAE-SPE HPLC) has been developed. A TM(010) microwave resonance cavity built in the laboratory was applied to concentrate the microwave energy. The sample placed in the zone of microwave irradiation was extracted with 95% acetonitrile (ACN) aqueous solution which was driven by a peristaltic pump at a flow rate of 1.0 mL min(-1). The extraction can be completed in a recirculating system in 10 min. When a number of extraction cycles were completed, the extract (1 mL) was diluted on-line with water. Then the extract was loaded into an SPE column where the analytes were retained while the unretained matrix components were washed away. Subsequently, the analytes were automatically transferred from the SPE column to the analytical column and determined by UV detector at 238 nm. The technique was used for determination of organochlorine pesticides (OCPs) in grains, including wheat, rice, corn and bean. The limits of detection of OCPs are in the range of 19-37 ng g(-1). The recoveries obtained by analyzing the four spiked grain samples are in the range of 86-105%, whereas the relative standard deviation (R.S.D.) values are <8.7% ranging from 1.2 to 8.7%. Our method was demonstrated to be fast, accurate, and precise. In addition, only small quantities of solvent and sample were required. PMID- 17418188 TI - Multivariate pattern recognition of petroleum-based accelerants by solid-phase microextraction gas chromatography with flame ionization detection. AB - A novel method has been developed for the extraction, analysis and identification of petroleum-based fuels using solid-phase microextraction with analysis by GC FID. Multivariate data analysis is employed to simplify these data allowing for more accurate classification. Principal component analysis (PCA) and soft independent modeling of class analogy (SIMCA) are explored for their effectiveness in establishing accelerant groupings based on the current and previous ASTM International guidelines. The SIMCA models developed for the previous and current ASTM system were 98.5% and 97.2% accurate in unknown sample class prediction. SPME in conjunction with multivariate data analysis is a new approach in accelerant sampling and classification. PMID- 17418189 TI - Anodic stripping voltammetry of antimony using gold nanoparticle-modified carbon screen-printed electrodes. AB - Carbon screen-printed electrodes (CSPE) modified with gold nanoparticles present an interesting alternative in the determination of antimony using differential pulse anodic stripping voltammetry. Metallic gold nanoparticles deposits have been obtained by direct electrochemical deposition. Scanning electron microscopy measurements show that the electrochemically synthesized gold nanoparticles are deposited in aggregated form. Any undue effects caused by the presence of foreign ions in the solution were also analyzed to ensure that common interferents in the determination of antimony by ASV. The detection limit for Sb(III) obtained was 9.44 x 10(-10) M. In terms of reproducibility, the precision of the above mentioned method in %R.S.D. values was calculated at 2.69% (n=10). The method was applied to determine levels of antimony in seawater samples and pharmaceutical preparations. PMID- 17418190 TI - Application of permeation liquid membrane and scanned stripping chronopotentiometry to metal speciation analysis of colloidal complexes. AB - The potential of permeation liquid membrane (PLM) to obtain dynamic metal speciation information for colloidal complexes is evaluated by measurements of lead(II) and copper(II) complexation by carboxyl modified latex nanospheres of different radii (15, 35, 40 and 65 nm). The results are compared with those obtained by a well characterized technique: stripping chronopotentiometry at scanned deposition potential (SSCP). Under the PLM conditions employed, and for large particles or macromolecular ligands, membrane diffusion is the rate limiting step. That is, the flux is proportional to the free metal ion concentration with only a small contribution from labile complexes. In the absence of ligand aggregation in the PLM channels, good agreement was obtained between the stability constants determined by PLM and SSCP for both metals. PMID- 17418191 TI - Excretion profile of boldenone and its metabolites after oral administration to veal calves. AB - The residue profiles of boldenone (17beta-Bol), its epimer (17alpha-Bol) and the related compound androsta-1,4-diene-3,17-dione (ADD), were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in urine of male calves orally treated with boldenone, boldenone esters, and/or ADD. In all the experiments with the administered steroids residues of 17alpha-Bol decreased rapidly after end of treatment; detectable amounts of 17alpha-Bol were however noticed along the withdrawal observation period after end of treatment. Differently, residues of 17beta-Bol were detectable only shortly after administration. This in vivo research concerning oral treatments of cattle with boldenone related substances proves ADD to be a very active boldenone precursor in bovine animals. PMID- 17418192 TI - Prolactin receptor and proliferating/apoptotic cells in esophagus of the Mozambique tilapia (Oreochromis mossambicus) in fresh water and in seawater. AB - We have previously shown that esophageal epithelium of a euryhaline goby displays elevated cell proliferation in freshwater (FW) fish, but undergo apoptosis during seawater (SW) acclimation. Prolactin (PRL) injection into the goby induced the cell proliferation, whereas cortisol treatment stimulated the cell proliferation and apoptosis [Takahashi, H., Sakamoto, T., Narita, K., 2006a. Cell proliferation and apoptosis in the anterior intestine of an amphibious, euryhaline mudskipper (Periophthalmus modestus). J. Comp. Physiol. B 176, 463-468, 2006). In the euryhaline tilapia (Oreochromis mossambicus), the dynamics of changes in cortisol glucocorticoid receptors (GR) during acclimation to different salinities also suggests a role for glucocorticoid signaling in the esophageal cell turnover, but the mode of PRL action remains largely unclear. In the present study, we report on effects in the tilapia esophagus that result from changes in environmental salinity. Specifically, we assessed: (1) mRNA expression of PRL receptor (PRLR) using quantitative real-time RT-PCR; (2) esophageal cell proliferation and apoptosis, using immunohistochemistry of proliferating cells nuclear antigen (PCNA) and in situ nick end-labeling of genomic DNA (TUNEL); (3) the possible localization of immunoreactive PRLR on proliferating/apoptotic cells. Plasma PRL increased after FW acclimation; PRLR mRNA levels in the esophagus of FW fish were significantly higher than those in SW-acclimated fish. Cell proliferation was induced randomly throughout the esophageal epithelium after FW acclimation, while cell division and increased apoptosis were concentrated at the tips of esophageal epithelial folds in SW-acclimated fish. Immunoreactive PRLR appeared to be localized at proliferating cells and at certain apoptotic cells, whereas immunoreactive GR was observed over the whole epithelium including the apoptotic/proliferating cells. Thus, PRL appears to affect cell turnover directly in the esophageal epithelium of the euryhaline tilapia. PMID- 17418193 TI - Postnatal depression and the experience of South Asian marriage migrant women in Taiwan: survey and semi-structured interview study. AB - OBJECTIVES: To investigate the experience of South Asian marriage migrant women in Taiwan by (1) determining their experiences and health beliefs during the postnatal period and their uptake of health care and determining the number of women with positive screening scores for postnatal depression during this period. METHODS: South Asian marriage migrant women were recruited using the following criteria: (1) Vietnamese and Indonesian women living in Taiwan for more than 1 year; (2) 6 months post-uncomplicated delivery; (3) conversational ability in Taiwanese and/or Mandarin Chinese and (4) a semi-structured interview and translated versions of the Edinburgh Postnatal Depression Scale (EPDS) investigated their experiences and health beliefs and identified the women with positive screens for postnatal depression. RESULTS: A total of 106 migrant women agreed to take part. Ninety-two women were from Vietnam (86.7%) and 14 from Indonesia (13.3%). More than half (56.6%) were aged between 19 and 24 (66% primigravida). The husbands of the women were mainly unskilled workers (80%). The frequency of a positive screen for postnatal depression was 25.5% (EPDS score >13). There was a statistically significant association between a score of 13 or more on the EPDS and self-reported 'emotional distress' following birth (p<0.01 ANOVA). Thematic analysis derived four key themes related to their experience and health beliefs: (1) Self-reported health: Although the main concern of the migrant women was the health of their baby and their families, some of them found their new families were unsupportive when they themselves were unwell. (2) Health beliefs: Dietary beliefs were important with a number of issues arising from a difference in health beliefs between themselves and their new families. (3) Access to health care: Communication problems and difficulties arising from their relationships with their new families affected 1/3 of women's access to health care and available health information. (4) Adaptation to a new environment: Many of the women described how difficult it was to adapt to their new environment and how they coped with the difficulties. CONCLUSIONS: South Asian migrant women in Taiwan face considerable challenges. If the health needs of these women are to be met, interventions which are "culturally competent" at the level of the individual, family and organisation need to be developed. PMID- 17418194 TI - Service user views and expectations of UK-registered mental health nurses: a systematic review of empirical research. AB - OBJECTIVES: To review national (UK) literature in order to (i) examine service user and carer views of UK-registered mental health nurses; (ii) identify the diversity of populations from which these views have been collected; (iii) assess the methodological rigour of the current knowledge base and (iv) evaluate the extent to which service users and carers have been involved in the development and execution of this work. This paper reports only on service users' views. DESIGN: Systematic review. DATA SOURCES: Electronic and evidence-based databases, reference checking and hand searching of key academic journals, national policy and user/carer organisational websites. REVIEW METHODS: Two reviewers independently undertook study eligibility judgements and data extraction. Eligible studies were sub-classified according to service setting (inpatient/residential, community/non-residential or mixed/unspecified). Each study was assessed against key quality criteria. Data were synthesised in a narrative format. RESULTS: One hundred and thirty two studies were included in the review. The majority were small-scale academic studies biased towards white, adult service users. Few studies provided evidence of user collaboration. Service users regard mental health nursing as a multi-faceted role delivering practical and social support alongside more formal psychological therapies. Service users report inadequate information provision, poor inter-professional communication and a lack of opportunities for collaborative care. Service users perceive inpatient mental health nurses as particularly inaccessible. CONCLUSIONS: UK registered mental health nurses should be equipped with both therapeutic clinical skills and generic skills associated with relationship building, engagement and communication. Future research should be conducted in collaboration with service users and include clear and effective mechanisms for the dissemination and implementation of research findings. In particular, the views of children and adolescents, the elderly and black and minority ethnic groups, currently under represented in research, should be examined. PMID- 17418201 TI - IFFS surveillance 07. PMID- 17418218 TI - Can long-term antibiotic treatment prevent progression of peripheral arterial occlusive disease? A large, randomized, double-blinded, placebo-controlled trial. AB - PURPOSE: The purpose was to investigate in a large, randomized, double-blinded, placebo-controlled trial, whether antibiotic treatment can prevent progression of peripheral arterial disease (PAD). MATERIAL AND METHODS: Five hundred and seven patients were included; all patients had an established diagnosis of PAD. Their mean age was 66 years (36-85), and 59% were males. Patients were randomized to Roxithromycin 300 mg daily for 28 days. Baseline investigations were ankle blood pressure, ankle-brachial blood pressure index (ABPI), walking distance, C. pneumoniae serology, cholesterol and medical history. Follow-up was performed every 6 months. Primary events were defined as death, peripheral revascularization and major lower limb amputation. Secondary events were thrombosis, stroke, transient cerebral ischaemic attack and myocardial infarction. Change in ABPI was also investigated. Data were analyzed mainly by Cox regression and linear regression. RESULTS: Included patients with PAD were randomized. Two patients withdrew. Of the remaining, 248 received roxithromycin and 257 placebo. In the treatment group 55% were seropositive and 53% in the placebo group. Mean follow-up was 2.1 years (range 0.06-5.1 years). In the placebo group, 26 died and 80 primary events occurred in total. In the treatment group, 28 died and 74 primary events were observed. The hazard ratio of death was 1.13 (95% CI: 0.68; 1.90), and of primary events 0.92 (95% CI: 0.67; 1.26). Also on secondary events and ABPI changes, no significant differences were found. CONCLUSION: Long-term treatment with roxithromycin is ineffective in preventing death, amputation, peripheral revascularization, myocardial infarction, stroke, transient cerebral ischaemic attack, thrombosis and decline in ABPI in patients with an established diagnosis of PAD. PMID- 17418219 TI - A comparison of the influence of global functional loads vs. local contact anatomy on articular cartilage thickness at the knee. AB - Cartilage contact geometry, along with joint loading, can play an important role in determining local articular cartilage tissue stress. Thus individual variations in cartilage thickness can be associated with both individual variations in joint loading associated with activities of daily living as well as individual differences in the anatomy of the contacting surfaces of the joint. The purpose of this study was to isolate the relationship between cartilage thickness predicted by individual variations in contact surface geometry based on the radii of the femur and tibia vs. cartilage thickness predicted by individual variations in joint loading. Knee magnetic resonance (MR) images and the peak knee adduction moments during walking were obtained from 11 young healthy male subjects (age 30.5+/-5.1 years). The cartilage thicknesses and surface radii of the femoral and tibial cartilage were measured in the weight-bearing regions of the medial and lateral compartments of three-dimensional models from the MR images. The ratio of contact pressure between the medial and lateral compartments was calculated from the radii of tibiofemoral contact surface geometries. The results showed that the medial to lateral pressure ratios were not correlated with the medial to lateral cartilage thickness ratios. However, in general, pressure was higher in the lateral than medial compartments and cartilage was thicker in the lateral than medial compartments. The peak knee adduction moment showed a significant positive linear correlation with medial to lateral thickness ratio in both femur (R(2)=0.43,P<0.01) and tibia (R(2)=0.32,P<0.01). The results of this study suggest that the dynamics of walking is an important factor to describe individual differences in cartilage thickness for normal subjects. PMID- 17418220 TI - Restricted-access material-based high-molecular-weight protein depletion coupled on-line with nano-liquid chromatography-mass spectrometry for proteomics applications. AB - Proteomics samples often contain both abundant proteins and low-level proteins and peptides. Highly abundant proteins can mask and/or bind those of lower abundance and thereby hinder their analysis. In particular, we were concerned with samples containing large amounts of albumin (up to 4.0 microM). In this study, a novel set-up for multidimensional nano-liquid chromatography-mass spectrometry (nanoLC-MS) with three columns coupled on-line was developed and characterised. A 1-mm-I.D. restricted-access-material (RAM) cartridge and a 100 microm-I.D. reversed-phase trap column are coupled in forward-flush mode to remove albumin before on-line separation on a 50 microm I.D. reversed-phase capillary analytical column. Volumes up to 100 microL of a complex matrix (containing 0.4 or 4.0 microM albumin) could be injected onto this system, enabling a 5000-fold volume reduction. Up to 99.7% of the albumin present in samples could be efficiently removed over the RAM cartridge. The total analysis time was about 40 min. Using Substance P as a model peptide, separations were efficient, with a peak width of 10s at half height. Moreover, separations were highly reproducible (relative standard deviation (RSD) on retention time approximately 3% over 1 week). The set-up proved to be robust and was used for about 750 analyses without exchanging one of the columns. Flexibility with respect to the stationary phase material in the sample preparation cartridge allows for other separation modes to be applied as well. PMID- 17418221 TI - Modeling of size-exclusion chromatography of electrolytes on non-ionic nanoporous adsorbents. AB - A dynamic model has been developed for chromatographic separation of mixed electrolyte solutions with non-ionic nanoporous adsorbents. The thermodynamic equilibrium condition at the pore entrance is written in terms of mixing, electrostatic and size-exclusion effects. The model is tested against experimental data measured with three binary mixtures on hypercrosslinked polystyrene and nanoporous carbon. The selectivity of the nanoporous adsorbents can be explained by the size-exclusion of the electrolytes and enrichment of both electrolytes in frontal chromatographic runs can be correlated satisfactorily with the proposed model. The model is also used to demonstrate continuous separation in a simulated moving-bed (SMB) system. PMID- 17418222 TI - Quality control techniques for routine analysis with liquid chromatography in laboratories. AB - The process of quality assurance should demonstrate that the method and the analytical instrument provide accurate and precise results, or whether deterioration occurs. For this, the quality procedure should include tests which provide information on the characteristic performance of the method. According to the recent literature, useful procedures contributing to the overall quality of analytical results are illustrated in relation with liquid chromatography. Parameters examined are carry-over, column statement, accuracy and precision. The performance of the whole system from the extraction to measurement of analytes of interest must be monitored with control charts. Data representing the routine performance of chromatographic equipment systems and method allow predicting long term uncertainties and confidence intervals. PMID- 17418223 TI - Chemometric analysis of complex hyphenated data. Improvements of the component detection algorithm. AB - Component detection algorithm (CODA) is a method to quickly extract the high quality mass chromatograms from complex liquid chromatography/electrospray ionization mass spectrometry (LC/MS) data sets, saving operators hours of analysis time. It appeared, however, that mass chromatograms with a limited baseline problem were ignored. This paper describes several methods to increase the tolerance for mass chromatograms with a baseline. PMID- 17418224 TI - Continuous full filling capillary electrochromatography: chromatographic performance and reproducibility. AB - Continuous full filling capillary electrochromatography with nanoparticles as pseudostationary phase interfaced with electrospray ionisation mass spectrometric detection was used for reversed phase separations with very high separation efficiency. Several batches of nanoparticles were synthesised and their electrochromatographic performance were evaluated. Different parameters, such as repeatability, reproducibility, limit of detection, and peak asymmetry, were investigated yielding excellent results. The stability of the system over wide pH ranges and over time was found to be excellent. Very high separation efficiencies with over 1.1 million theoretical plates per metre were obtained. The limit of detection for the investigated dialkyl phthalates was approximately 1.0 micromol L(-1), corresponding to 3-5 fmol injected. After preparation, nanoparticle suspensions could be used without further treatment for at least an entire working day with maintained chromatographic qualities. PMID- 17418225 TI - Automated compound classification for ambient aerosol sample separations using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry. AB - Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GCxGC-TOF-MS) is useful in analyzing complex samples, such as the semi-volatile organic compounds (SVOC) in daily aerosol samples collected in Augsburg, Germany. For efficient routine analysis, a partial classification system based on mass spectra has been proposed to group compounds into substance classes. Classifiers based on fragmentation patterns, retention time, and spectral transformations are incorporated into software scripts for automated classification. Fragmentation pattern rules are also applied to an aerosol sample analyzed using direct thermal desorption (DTD)-GCxGC-TOF-MS and a non-traditional column combination. PMID- 17418226 TI - Development of an absolute quantification method targeting growth hormone biomarkers using liquid chromatography coupled to isotope dilution mass spectrometry. AB - A method to perform absolute quantification of two biomarkers (IGF-1 and IGFBP-3) of growth hormone abuse has been developed. Isotope dilution is used with synthetically labelled peptides as internal standards. Peptide selection and multiple reaction monitoring design are discussed. A simple sample preparation based on the reduction and alkylation of cysteine residues followed by tryptic digestion provides a sufficient digestion of proteins. Serum samples fortified with increasing amounts of target proteins are analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) on a triple quadrupole mass spectrometer. Specificity is ensured by the selection of sequences with no homology in BLAST, as well as retention time deviation check, and ion ratio monitoring. Linearity is studied in terms of calibration curves. These curves for IGFBP-3 and IGF-1 are generated with mean slopes of 0.055 and 0.065, intercepts of 0.107 and -0.011, and with coefficients of correlation of 0.95 and 0.98, respectively. These curves result from the addition of proteins to the serum. Risks of variations related to potential matrix effects are therefore reduced, as well as probable variations related to the digestion steps. The working concentration ranges are 4-10 ng/microl for IGFBP-3 and 2-8 ng/microl for IGF-1. Preliminary data regarding repeatability show that relative standard deviations (RSDs) range between 13 and 32% for IGFBP-3 and between 7 and 29% for IGF-1. PMID- 17418227 TI - Efficiency enhancements in micellar liquid chromatography through selection of stationary phase and alcohol modifier. AB - Micellar liquid chromatography (MLC) remains hindered by reduced chromatographic efficiency compared to reversed phase liquid chromatography (RPLC) using hydro organic mobile phases. The reduced efficiency has been partially explained by the adsorption of surfactant monomers onto the stationary phase, resulting in a slow mass transfer of the analyte within the interfacial region of the mobile phase and stationary phase. Using an array of 12 columns, the effects of various bonded stationary phases and silica pore sizes, including large-pore short alkyl chain, non-porous, superficially porous and perfluorinated, were evaluated to determine their impact on efficiency in MLC. Additionally, each stationary phase was evaluated using 1-propanol and 1-butanol as separate micellar mobile phase alcohol additives, with several columns also evaluated using 1-pentanol. A simplified equation for calculation of A' and C' terms from reduced plate height (h) versus reduced velocity (nu) plots was used to compare the efficiency data obtained with the different columns and mobile phases. Analyte diffusion coefficients needed for the h versus nu plots were determined by the Taylor-Aris dispersion technique. The use of a short alkyl chain, wide-pore silica column, specifically, Nucleosil C4, 1000A, was shown to have the most improved efficiency when using a micellar mobile phase compared to a hydro-organic mobile phase for all columns evaluated. The use of 1-propanol was also shown to provide improved efficiency over 1-butanol or 1-pentanol in most cases. In a second series of experiments, column temperatures were varied from 40 to 70 degrees C to determine the effect of temperature on efficiency for a subset of the stationary phases. Efficiency improvements ranging from 9% for a Chromegabond C8 column to 58% for a Zorbax ODS column were observed over the temperature range. Based on these observed improvements, higher column temperatures may often yield significant gains in column efficiency, assuming the column is thermally stable. PMID- 17418228 TI - Shear-induced structures formed during thixotropic loops in dilute worm-micelle solutions. AB - The shear stress response of the system CTAB/NaSal/water is studied using strain thixotropic loops in the low concentration regime and [NaSal]/[CTAB]=1. Stress response during up-shear curves depends on the ramping rate. However, stress response during down-shear curves collapses in one curve, no matter the rate used during the ramping down process; it does not depend on the history of the system. In the process of ramping up during the thixotropic loops, the system forms shear induced structures. We were able to observe them through the scattered light produced by those structures, when the fluid is under shear in the gap of a transparent Couette cell. During the ramping down, the shear-induced structures survive until the shear rate vanishes. Models that describe the form of the up shear and down-shear curves were presented, as well as, how thixotropic loops can give information about this kind of systems. In particular, for down-shear curves, we can define a decay shear rate constant that follows an Arrhenius temperature dependence. PMID- 17418229 TI - Heterodimeric amino acid transporters in the placenta--a workshop report. PMID- 17418230 TI - The role of central nervous system plasticity in tinnitus. AB - Tinnitus is a vexing disorder of hearing characterized by sound sensations originating in the head without any external stimulation. The specific etiology of these sensations is uncertain but frequently associated with hearing loss. The "neurophysiogical" model of tinnitus has enhanced appreciation of central nervous system (CNS) contributions. The model assumes that plastic changes in the primary and non-primary auditory pathways contribute to tinnitus with the former perhaps sustaining them, and the latter contributing to perceived severity and emotionality. These plastic changes are triggered by peripheral injury, which results in new patterns of brain activity due to anatomic alterations in the connectivity of CNS neurons. These alterations may change the balance between excitatory and inhibitory brain processes, perhaps producing cascades of new neural activity flowing between brainstem and cortex in a self-sustaining manner that produces persistent perceptions of tinnitus. The bases of this model are explored with an attempt to distinguish phenomenological from mechanistic explanations. LEARNING OUTCOMES: (1) Readers will learn that the variables associated with the behavioral experience of tinnitus are as complex as the biological variables. (2) Readers will understand what the concept of neuroplastic brain change means, and how it is associated with tinnitus. (3) Readers will learn that there may be no one brain location associated with tinnitus, and it may result from interactions between multiple brain areas. (4) Readers will learn how disinhibition, spontaneous activity, neural synchronization, and tonotopic reorganization may contribute to tinnitus. PMID- 17418231 TI - Early middle ear effusion and language at age seven. AB - This study examined the relation of middle ear effusion (MEE) in the first 3 years of life to language outcomes at age seven. It was hypothesized, on the basis of a literature review, that (1) a low, but positive relation between early MEE and language measures in general will be observed at age seven, and (2) major effects will be demonstrated for measures of articulation and phonological sensitivity. MEE was assessed as days with bilateral MEE and episodes of MEE. Three measures of language status were used: the Test of Auditory Analysis Skill (TAAS [Rosner, J. (1975). TAAS: Test of Auditory Analysis Skill. Novato, CA: Academic Therapy Publications]), Goldman-Fristoe Articulation Test, Sounds in Words and Sounds in Sentences (GFAT [Goldman, R., & Fristoe, M. (1986). Goldman Fristoe Test of Articulation. Circle Pines, MN: American Guidance Service]), and the Clinical Evaluation of Language Fundamentals-revised (CELF-R [Semel, E. M., Wiig, E. H., & Secord, W. (1987). CELF: Clinical Evaluation of Language Fundamentals-Revised. New York: Harcourt Brace Jovanovich]). The sample included 179 children who were heterogeneous for SES and ethnicity. There were no significant correlations for MEE and language measures. These negative results were sustained when multiple regression was used with controls for socioeconomic status and quality of the home environment. LEARNING OUTCOMES: We conclude that early MEE may not pose a threat to language development in the early school years. PMID- 17418232 TI - Shape and subunit organisation of the DNA methyltransferase M.AhdI by small-angle neutron scattering. AB - Type I restriction-modification (R-M) systems encode multisubunit/multidomain enzymes. Two genes (M and S) are required to form the methyltransferase (MTase) that methylates a specific base within the recognition sequence and protects DNA from cleavage by the endonuclease. The DNA methyltransferase M.AhdI is a 170 kDa tetramer with the stoichiometry M(2)S(2) and has properties typical of a type I MTase. The M.AhdI enzyme has been prepared with deuterated S subunits, to allow contrast variation using small-angle neutron scattering (SANS) methods. The SANS data were collected in a number of (1)H:(2)H solvent contrasts to allow matching of one or other of the subunits in the multisubunit enzyme. The radius of gyration (R(g)) and maximum dimensions (D(max)) of the M subunits in situ in the multisubunit enzyme (50 A and 190 A, respectively) are close of those of the entire MTase (51 A and 190 A). In contrast, the S subunits in situ have experimentally determined values of R(g)=35 A and D(max)=110 A, indicating their more central location in the enzyme. Ab initio reconstruction methods yield a low resolution structural model of the shape and subunit organization of M.AhdI, in which the Z-shaped structure of the S subunit dimer can be discerned. In contrast, the M subunits form a much more elongated and extended structure. The core of the MTase comprises the two S subunits and the globular regions of the two M subunits, with the extended portion of the M subunits most probably forming highly mobile regions at the outer extremities, which collapse around the DNA when the MTase binds. PMID- 17418233 TI - Evidence for a novel binding site conformer of aldose reductase in ligand-bound state. AB - Human aldose reductase (ALR2) has evolved as a promising therapeutic target for the treatment of diabetic long-term complications. The binding site of this enzyme possesses two main subpockets: the catalytic anion-binding site and the hydrophobic specificity pocket. The latter can be observed in the open or closed state, depending on the bound ligand. Thus, it exhibits a pronounced capability for induced-fit adaptations, whereas the catalytic pocket exhibits rigid properties throughout all known crystal structures. Here, we determined two ALR2 crystal structures at 1.55 and 1.65 A resolution, each complexed with an inhibitor of the recently described naphtho[1,2-d]isothiazole acetic acid series. In contrast to the original design hypothesis based on the binding mode of tolrestat (1), both inhibitors leave the specificity pocket in the closed state. Unexpectedly, the more potent ligand (2) extends the catalytic pocket by opening a novel subpocket. Access to this novel subpocket is mainly attributed to the rotation of an indole moiety of Trp 20 by about 35 degrees . The newly formed subpocket provides accommodation of the naphthyl portion of the ligand. The second inhibitor, 3, differs from 2 only by an extended glycolic ester functionality added to one of its carboxylic groups. However, despite this slight structural modification, the binding mode of 3 differs dramatically from that of the first inhibitor, but provokes less pronounced induced-fit adaptations of the binding cavity. Thus, a novel binding site conformation has been identified in a region where previous complex structures suggested only low adaptability of the binding pocket. Furthermore, the two ligand complexes represent an impressive example of how the slight change of a chemically extended side-chain at a given ligand scaffold can result in a dramatically altered binding mode. In addition, our study emphasizes the importance of crystal structure analysis for the translation of affinity data into structure-activity relationships. PMID- 17418234 TI - Modulation of the antigenic peptide transporter TAP by recombinant antibodies binding to the last five residues of TAP1. AB - The transporter associated with antigen processing (TAP) plays a pivotal role in the major histocompatibility complex (MHC) class I mediated immune response against infected or malignantly transformed cells. It belongs to the ATP-binding cassette (ABC) superfamily and consists of TAP1 (ABCB2) and TAP2 (ABCB3), each of which possesses a transmembrane and a nucleotide-binding domain (NBD). Here we describe the generation of recombinant Fv and Fab antibody fragments to human TAP from a hybridoma cell line expressing the TAP1-specific monoclonal antibody mAb148.3. The epitope of the antibody was mapped to the very last five C-terminal amino acid residues of TAP1 on solid-supported peptide arrays. The recombinant antibody fragments were heterologously expressed in Escherichia coli and purified to homogeneity from periplasmic extracts by affinity chromatography. The monoclonal and recombinant antibodies bind with nanomolar affinity to the last five C-terminal amino acid residues of TAP1 as demonstrated by ELISA and surface plasmon resonance. Strikingly, the recombinant antibody fragments confer thermal stability to the heterodimeric TAP complex. At the same time TAP is arrested in a peptide transport incompetent conformation, although ATP and peptide binding to TAP are not affected. Based on our results we suggest that the C terminus of TAP1 modulates TAP function presumably as part of the dimer interface of the NBDs. PMID- 17418235 TI - Molecular determinants of antibiotic recognition and resistance by aminoglycoside phosphotransferase (3')-IIIa: a calorimetric and mutational analysis. AB - The growing threat from the emergence of multidrug resistant pathogens highlights a critical need to expand our currently available arsenal of broad-spectrum antibiotics. In this connection, new antibiotics must be developed that exhibit the abilities to circumvent known resistance pathways. An important step toward achieving this goal is to define the key molecular interactions that govern antibiotic resistance. Here, we use site-specific mutagenesis, coupled with calorimetric, NMR, and enzymological techniques, to define the key interactions that govern the binding of the aminoglycoside antibiotics neomycin and kanamycin B to APH(3')-IIIa (an antibiotic phosphorylating enzyme that confers resistance). Our mutational analyses identify the D261, E262, and C-terminal F264 residues of the enzyme as being critical for recognition of the two drugs as well as for the manifestation of the resistance phenotype. In addition, the E160 residue is more important for recognition of kanamycin B than neomycin, with mutation of this residue partially restoring sensitivity to kanamycin B but not to neomycin. By contrast, the D193 residue partially restores sensitivity to neomycin but not to kanamycin B, with the origins of this differential effect being due to the importance of D193 for catalyzing the phosphorylation of neomycin. These collective mutational results, coupled with (15)N NMR-derived pK(a) and calorimetrically derived binding-linked drug protonation data, identify the 1-, 3 , and 2'-amino groups of both neomycin and kanamycin B as being critical functionalities for binding to APH(3')-IIIa. These drug amino functionalities represent potential sites of modification in the design of next-generation compounds that can overcome APH(3')-IIIa-induced resistance. PMID- 17418237 TI - Inflammation versus neurodegeneration: consequences for treatment. AB - Inflammation is assumed to be the main driver of neurodegeneration in multiple sclerosis. However there is evidence that questions this hypothesis: Data from pathological studies and from imaging have shown that neuronal/axonal damage occurs early in lesion formation and at the earliest clinical stage of the disease. Additionally axonal damage itself can lead to inflammation and laboratory and clinical observations suggest that inflammation may be neuroprotective. Finally, clinical trials reveal that immunosuppression does not have the predicted effect on axonal damage and disability. Clearly if inflammation is not the primary event but a secondary phenomenon this has important implications in developing treatment regimes. PMID- 17418236 TI - Identification and characterization of CMP-NeuAc:GalNAc-IgA1 alpha2,6 sialyltransferase in IgA1-producing cells. AB - Glycosylation defects occur in several human diseases. In IgA nephropathy, IgA1 contains O-glycans that are galactose-deficient and consist mostly of core 1 alpha2,6 sialylated N-acetylgalactosamine, a configuration suspected to prevent beta1,3 galactosylation. We confirmed the same aberrancy in IgA1 secreted by the human DAKIKI B cell line. Biochemical assays indicated CMP-NeuAc:GalNAc-IgA1 alpha2,6-sialyltransferase activity in this cell line. However, a candidate enzyme, ST6-GalNAcI, was not transcribed in DAKIKI cells, B cells isolated from blood, or Epstein-Barr virus (EBV)-immortalized IgA1-producing cells from the blood of IgAN patients and healthy controls. Instead, ST6-GalNAcII transcription was detected at a high level. Expression of the ST6-GalNAcII gene and activity of the CMP-NeuAc:GalNAc-IgA1 alpha2,6-sialyltransferase were higher in IgA1 producing cell lines from IgAN patients than in such cells from healthy controls. These data are the first evidence that human cells that lack ST6-GalNAcI can sialylate core 1 GalNAc-Ser/Thr. PMID- 17418238 TI - A probabilistic model of reserve design. AB - We develop a probabilistic approach to optimum reserve design based on the species-area relationship. Specifically, we focus on the distribution of areas among a set of reserves maximizing biodiversity. We begin by presenting analytic solutions for the neutral case in which all species have the same colonization probability. The optimum size distribution is determined by the local-to-regional species richness ratio k. There is a critical k(t) ratio defined by the number of reserves raised to the scaling exponent of the species-area relationship. Below k(t), a uniform area distribution across reserves maximizes biodiversity. Beyond k(t), biodiversity is maximized by allocating a certain area to one reserve and uniformly allocating the remaining area to the other reserves. We proceed by numerically exploring the robustness of our analytic results when departing from the neutral assumption of identical colonization probabilities across species. PMID- 17418239 TI - Effect of DL-alpha-lipoic acid on cyclophosphamide induced lysosomal changes in oxidative cardiotoxicity. AB - Cyclophosphamide (CP), one of the widely prescribed antineoplastic drugs can cause fatal cardiotoxicity. The present study is aimed at evaluating the cardioprotective role of lipoic acid in CP induced toxicity. Male albino rats of Wistar strain were divided into four groups and treated as follows: Group I served as control, Group II received a single dose of CP (200 mg/kg b.wt., i.p.), Group III received lipoic acid (25 mg/kg b.wt., orally) for 10 days, and Group IV received CP immediately followed by lipoic acid for 10 days. In CP administered rats, the levels of protein carbonyl and 8-hydroxy-2-deoxyguanosine were increased significantly (P<0.001) indicating oxidative changes in the heart tissue. The activities of lysosomal acid hydrolases, beta-Glu, beta-Gal, NAG, Cat D and ACP increased significantly (P<0.001) in the serum as well as in the heart tissue after CP administration. An increase in hydroxyproline was observed in CP induced rats. Lipoic acid effectively reverted these abnormal biochemical changes to near normalcy. These observations highlight the protective role of lipoic acid in CP induced cardiotoxicity. PMID- 17418240 TI - A nutrient loading budget for Biscayne Bay, Florida. AB - The water quality in Biscayne Bay has been significantly affected by past and continuing coastal and watershed development. The nutrient concentrations in the Bay have been dramatically changed by the conversion of natural creeks and sheet flow freshwater inputs to rapid and episodic canal inputs from the large and rapidly expanding Miami metropolitan area. This study is an evaluation of nutrient loadings to Biscayne Bay for 1994-2002 from canal, atmospheric, and groundwater sources. Dissolved inorganic nitrogen (DIN, as nitrate, nitrite, and ammonium) and total phosphorus (TP) loadings by the canals were influenced by their geographic locations relative to discharge amount, watershed land use, stormwater runoff, and proximity to landfills. Annual budgets showed that canals contributed the bulk of N loading to the bay as 1687.2 metric ton N yr(-1) (88% total load). Direct atmospheric DIN load for Biscayne Bay was only 231.7 ton N yr(-1), based on surface area. Of the canal DIN load, nitrate+nitrite (NO(x)(-)) loading (1294.5 ton N yr(-1)) made up a much greater proportion than that of ammonium (NH(4)(+), 392.6 ton N yr(-1)). In the urbanized north and central Bay, canal DIN load was evenly split between NO(x)(-) and NH(4)(+). However, in the south, 95% of the DIN load was in the form of NO(x)(-), reflecting the more agricultural land use. Contrary to N, canals contributed the only 66% of P load to the bay (27.5 ton P yr(-1)). Atmospheric TP load was 14 ton Pyr(-1). In the north, canal P load dominated the budget while in the south, atmospheric load was almost double canal load. Groundwater inputs, estimated only for the south Bay, represented an important source of N and P in this zone. Groundwater input of N (141 ton N yr(-1)) was about equal to atmospheric load, while P load (5.9 ton P yr(-1)) was about equal to canal load. PMID- 17418241 TI - Effect of chaotic vasomotion in skeletal muscle on tissue oxygenation. AB - Vasomotion refers to spontaneous variations in the lumen size of small vessels, with a plausible role in regulation of various aspects of microcirculation. We propose a computational model of vasomotion in skeletal muscle in which the pattern of vasomotion is shown to critically determine the efficiency of oxygenation of a muscle fiber. In this model, precapillary sphincters are modeled as nonlinear oscillators. We hypothesize that these sphincters interact via exchange of vasoactive substances. As a consequence, vasomotion is described as a phenomenon associated with a network of nonlinear oscillators. As a specific instance, we model the vasomotion of precapillary sphincters surrounding an active fiber. The sphincters coordinate their rhythms so as to minimize oxygen deficit in the fiber. Our modeling studies indicate that efficient oxygenation of the fiber depends crucially on the mode of interaction among the vasomotions of individual sphincters. While chaotic forms of vasomotion enhanced oxygenation, regular patterns of vasomotion failed to meet the oxygenation demand accurately. PMID- 17418242 TI - Chromosomal aberrations in environmentally exposed population in relation to metabolic and DNA repair genes polymorphisms. AB - The capital city of Prague is one of the most polluted localities of the Czech Republic. Therefore, the effect of exposure to carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) adsorbed onto respirable air particles (<2.5mum) on chromosomal aberrations was studied in a group of policemen (males, aged 22-50 years) working in the downtown area of Prague and spending daily >8h outdoors (N=53). Age- and sex-matched healthy volunteers spending >90% daily time indoors were chosen as controls (N=52). Ambient air particles (PM10, PM2.5) and c-PAHs were monitored using versatile air pollution sampler (VAPS), and personal exposure was evaluated using personal samplers during working shift. Chromosomal aberrations were analyzed by conventional cytogenetic analysis and fluorescent in situ hybridization (FISH). Urinary cotinine plasma levels of vitamins A, E and C, folate, total cholesterol, HDL, LDL cholesterols and triglycerides were also analyzed as possible effect modifiers. Genotypes CYP1A1*2A, CYP1A1*2C, GSTM1, GSTP1, GSTT1, EPHX1, NAT2, hOGG1, XRCC1, XPD, p53 BstI, p53 MspI, MTHFR677, and MS2656 were determined by PCR-based RFLP assays. The following levels of air pollution were recorded during the study period (mean from HiVol sampling): PM10 62.6microg/m(3), c-PAHs 24.7ng/m(3), B[a]P 3.50ng/m(3). The conventional cytogenetic analysis did not reveal any differences between the group of policemen exposed to the ambient air pollution and the control group. The cytogenetic analysis by FISH analysis used the whole chromosome painting probes for chromosomes #1 and #4 (Cambio, UK). It detected a significant increase in all studied endpoints in the policemen compared to controls (% AB.C.=0.33+/-0.25 versus 0.24+/-0.18, p<0.05, F(G)/100=1.72+/-1.57 versus 1.25+/-1.11, p<0.05, AB/1000 (aberrations/1000 cells)=5.58+/-4.62 versus 3.90+/-3.06, p<0.05). CYP1A1*2C (Ile/Ile), XPD 23 (Lys/Lys), and XPD 6 (CC) genotypes were associated with an increase of aberrant cells by conventional method. Factors associated with an increased level of translocations by FISH included age, smoking, B[a]P like DNA adducts (corresponding to the exposure of c-PAHs), folate, polymorphisms of CYP1A1*2C, GSTP1, EPHX1, p53 MspI and MTHFR. Ambient air exposure to c-PAHs significantly increased FISH cytogenetic parameters in nonsmoking policemen. We may conclude that FISH indicates that the city policemen in Prague represent a group of increased genotoxic risk. This is the first study that has reported a relationship between DNA adducts (biomarker of exposure) and chromosomal aberrations by FISH (biomarker of effect). PMID- 17418243 TI - Extracts from The Cochrane Library: Allergen injection immunotherapy for seasonal allergic rhinitis (review). PMID- 17418244 TI - Practical guide to analyzing your own practice: steps toward "Prospective analysis of practice". PMID- 17418245 TI - Current concepts in microvascular reconstruction. PMID- 17418246 TI - A systematic review of patient-reported outcome measures in head and neck cancer surgery. AB - OBJECTIVE: To identify, summarize, and evaluate patient-reported outcome questionnaires for use in head and neck cancer surgery with the view to making recommendations for future research. DATA SOURCES: A systematic review of the English-language literature, with the use of head-and-neck-surgery-specific keywords, was performed in the following databases: Medline, Embase, HAPI, CINAHL, Science/Social Sciences Citation Index, and PsycINFO from 1966 to March 2006. DATA EXTRACTION AND STUDY SELECTION: All English-language instruments identified as patient-reported outcome questionnaires that measure quality of life and/or satisfaction that had undergone development and validation in a head and neck cancer surgery population were included. DATA SYNTHESIS: Twelve patient reported outcome questionnaires fulfilled our inclusion criteria. Of these, four were developed from expert opinion alone or did not have a published development process and seven questionnaires lacked formal item reduction. Only three questionnaires (EORTC Head and Neck Module, University of Michigan Head and Neck Quality-of-life Questionnaire, and Head and Neck Cancer Inventory) fulfilled guidelines for instrument development and evaluation as outlined by the Medical Outcomes Trust. CONCLUSIONS: Rigorous instrument development is important for creating valid, reliable, and responsive disease-specific questionnaires. As a direction for future instrument development, an increased focus on qualitative research to ensure patient input may help to better conceptualize and operationalize the variables most relevant to head and neck cancer surgery patients. In addition, the use of alternative methods of psychometric data analysis, such as Rasch, may improve the value of health measurement in clinical practice for individual patients. PMID- 17418247 TI - Comorbidities in head and neck cancer: agreement between self-report and chart review. AB - OBJECTIVES: To determine the accuracy of self-reported comorbidities compared with medical record review and the clinical and sociodemographic characteristics associated with accuracy of self-reported comorbidities. STUDY DESIGN: We conducted a prospective study of 458 newly diagnosed head and neck cancer patients using self-administered questionnaire and medical chart review data. Overall and itemwise consistency between self-report and chart review was evaluated. Social, clinical, and demographic characteristics of consistent versus inconsistent responders were analyzed. RESULTS: Seventy-four percent of patients had at least one comorbidity. There was good overall consistency between self report and chart review (kappa = 0.50). Compared with consistent responders, inconsistent responders were found to be older (P < 0.05), have lower sleep (P < 0.05) and physical activity scores (P < 0.05), be more depressed (P < 0.05), and have more severe comorbidities (P < 0.05). CONCLUSIONS AND SIGNIFICANCE: Self report may be considered as an alternative to chart review for comorbidity assessment in head and neck cancer patients. Younger patients, those with good general health, fewer depressive symptoms, and mild comorbidities, are more likely to give responses consistent with chart review. PMID- 17418248 TI - Immunoscintigraphy of patients with head and neck carcinomas, with an anti angiogenetic antibody fragment. AB - OBJECTIVE: In a phase I/II clinical study, we investigated tumor targeting in patients with head and neck squamous cell carcinomas (SCC), using an antibody directed against the extra-domain-B of fibronectin (EDB), a marker of angiogenesis and tissue remodeling. STUDY DESIGN AND SETTING: Five patients with SCC were injected with the 123-iodine-radiolabeled L19(scFv)2 antibody and underwent scintigraphic detection with single photon emission tomography with computerized tomography (SPECT/CT). In addition, 18F-fluorodeoxyglucose (18FDG) positron emission tomography with computerized tomography (PET/CT) was performed. RESULTS: Successful targeting of the primary tumor could be achieved in 4 of 5 patients and was comparable to PET imaging. No side effects were observed. CONCLUSIONS: Tumor targeting with the L19(scFv)2 antibody is also feasible for head and neck SCC. SIGNIFICANCE: These results may serve as a base for future therapeutical applications in human beings, with modified versions of the L19(scFv)2 antibody, designed to selectively deliver bioactive molecules into malignant tumors. PMID- 17418249 TI - Accelerated radiotherapy with simultaneous integrated boost fractionation and intensity-modulated radiotherapy for advanced head and neck cancer. AB - OBJECTIVE: To determine the feasibility and toxicity profile of accelerated radiotherapy with a simultaneous integrated boost fractionation scheme with intensity-modulated radiotherapy (SIB-IMRT) with or without chemotherapy. STUDY DESIGN AND SETTING: Forty-nine patients with advanced head and neck cancer underwent SIB-IMRT. Concomitant chemotherapy was administered in 29 patients. RESULTS: Grade 3 acute toxicities included 55% mucositis, 20% odynophagia, 12% nausea, 18% hematologic, and 8% skin. There were no grade 4 toxicities or treatment-related deaths. With a median follow-up of 25 months, locoregional control was 83%, and overall survival was 80%. Of patients with grade 3 late toxicities, two patients (4% of the total) required a permanent percutaneous endoscopic gastrostomy tube, and osteonecrosis occurred in one patient (2% of the total). CONCLUSIONS: SIB-IMRT is a feasible technique that shortens the overall treatment time in the radical treatment of patients with advanced head and neck cancer while maintaining acceptable rates of acute toxicity in this study. Although the results are promising, this approach should be considered only in the setting of a clinical trial. PMID- 17418250 TI - Outpatient thyroid surgery is safe and desirable. AB - BACKGROUND: Thyroid surgery has traditionally been done on an inpatient basis. With the advent of minimal access techniques, drains are frequently not required and ambulatory thyroidectomy is possible. DESIGN: Prospective, nonrandomized analysis of consecutive series of patients. METHODS AND MATERIALS: Patients undergoing thyroid surgery between 12/1/04 and 10/31/05 were stratified based on admission status. Demographic data were collected and outcome measures were considered. RESULTS: Ninety-one patients underwent thyroid surgery. Fifty-two were done on an outpatient basis, 26 patients were observed under a 23-hour status, and 13 were admitted. There were two complications in the outpatient group and one in the inpatient group (P = 1.0). Costs were significantly lower for outpatients ($7,814) than for inpatients ($10,288; P < 0.0001). SIGNIFICANCE: In carefully selected patients who prefer convalescence at home, outpatient thyroidectomy can be performed safely and cost-effectively, particularly when prophylactic calcium supplementation is utilized after total thyroidectomy to prevent transient postoperative hypocalcemia. PMID- 17418251 TI - Increased incidence of papillary thyroid cancer among total thyroidectomies in Crete. AB - OBJECTIVE: To investigate the increased incidence of papillary thyroid cancer as found in specimens of total thyroidectomies and potential correlation with etiological factors. STUDY DESIGN AND SETTING: A retrospective study on patients who underwent total thyroidectomy, from 1990 to 2004, in an academic tertiary referral medical center. Patients' records were placed in a database, which included medical condition, history, and demographics. Histopathological slides were reviewed with special focus on papillary cancer. RESULTS: Our series consisted of 2379 patients. Thyroid cancer was confirmed in 354 patients (14.88%). Papillary carcinoma represented the most frequent type (316 patients, 89.26%). Increased incidence of papillary carcinomas was noticed after 1995, reaching the maximum value in the year 2000. After 2000, there was a descending trend and then a plateau. CONCLUSION: The increased incidence of papillary thyroid cannot be attributed to dietary patterns or increased diagnostic and therapeutic activity. It is likely to be associated with increased radiation and may be associated with the Chernobyl fallout. PMID- 17418252 TI - Injectable hyaluronic acid for the correction of HIV-associated facial lipoatrophy. AB - OBJECTIVE: To evaluate the use of Perlane, an injectable form of hyaluronic acid, for the correction of HIV-associated facial lipoatrophy. STUDY DESIGN AND SETTING: A prospective, observational study in a consecutive series of 18 HIV positive males with facial lipoatrophy injected with Perlane. Fourteen patients were available for final analysis. RESULTS: Mean follow-up time was 12 months. Based on photographic analysis, there was a significant early improvement (P = 0.0035). This difference remained significant after 12 months (P = 0.04666); no significant difference in grade was shown between 1 and 12 months (P = 0.3693). Office assessments of improvement showed an early marked improvement in 85.8% of patients, and 78.6% of subjects demonstrated at least moderate improvement at 12 months. CONCLUSION/SIGNIFICANCE: Patient satisfaction was high, with only minor side effects and no late complications. Our findings support Perlane to be a feasible option for the provisional correction of mild to moderate facial lipoatrophy. PMID- 17418253 TI - Endoscopic repair of anterior table: frontal sinus fractures with a Medpor implant. AB - OBJECTIVES: Isolated anterior table frontal sinus fractures are commonly repaired through a coronal incision. Endoscopic repair of these injuries has recently been described. This study evaluates the endoscopic repair of isolated anterior table frontal sinus fractures with a Medpor implant. METHODS: Preinjury photographs of 10 cadaveric heads were obtained. Anterior table frontal sinus fractures were generated in all cadavers. The fractures were documented with postinjury computerized tomography (CT) scans. The fractures were then endoscopically repaired with a Medpor implant. Five cadavers received prefabricated implants generated from the post injury CT data. Five cadavers received a standard implant (0.85 mm sheet) contoured intraoperatively. The success of each repair was documented with post repair CT scans, photographs, and direct transcutaneous visualization. RESULTS: All 10 defects were successfully repaired within 1 to 2 mm. All implants were palpable, but no objective asymmetry could be appreciated photographically or on CT scan. CONCLUSION: Anterior table frontal sinus fractures can be endoscopically repaired with either a standard 0.85 mm sheet or a prefabricated implant. Clinical application of this technique would be expected to reduce operating time, surgical morbidity, and cost. PMID- 17418254 TI - Clopidogrel (Plavix) reduces the rate of thrombosis in the rat tuck model for microvenous anastomosis. AB - OBJECTIVE: To evaluate the effect of clopidogrel on the rate of thrombosis in a rat model for venous microvascular failure. STUDY DESIGN AND SETTING: Forty rats were treated with clopidogrel or saline control via gastric gavage in a randomized, blinded fashion. After allowing for absorption and activation, each femoral vein was isolated and a venous "tuck" procedure was performed. The bleeding time and vessel patency were subsequently evaluated. RESULTS: The rate of vessel thrombosis was decreased in the clopidogrel-treated group compared to controls (7.9% vs 31.4%, P < 0.025). The bleeding time was longer in the clopidogrel-treated group compared to controls (250 +/- 100 seconds vs 173 +/- 59 seconds, P < 0.015). CONCLUSION: Clopidogrel decreased the rate of thrombosis in the rat model for venous microvascular failure. SIGNIFICANCE: The use of clopidogrel may reduce the rate of venous thrombosis after free tissue transfer and may be indicated in select patients. PMID- 17418255 TI - Biofilm formation in cochlear implants with cochlear drug delivery channels in an in vitro model. AB - BACKGROUND: Cochlear implant (CI) drug delivery (DD) may improve electrophysiological outcomes, but it may also increase the risk of suppurative complications. The aim of this study was to evaluate the development of bacterial biofilms on DD ports when subjected to varying types of penetration. METHODS: Silastic models were constructed to represent CIs with a DD channel, with an intact port, a widely opened port, a noncoring needle penetrating the port, and a noncoring needle removed from the port. CIs were exposed to a culture of a biofilm-forming strain of Staphylococcus aureus for 5 days. Biofilm formation was assessed with quantitative bacterial counts (after eliminating planktonic bacteria) and scanning electron microscopy. RESULTS: Bacterial counts were significantly higher in CIs with widely fenestrated ports than all other port conditions (P = 0.0003). CONCLUSIONS: Biofilm formation may be minimized on CIs with DD by using fine, noncoring needles and limiting the duration of port penetration. PMID- 17418256 TI - Biofilm formation in an in vitro model of cochlear implants with removable magnets. AB - BACKGROUND: Cochlear implant (CI) recesses, such as the removable magnet pocket, appear to harbor more biofilm than smooth surfaces. The aim of this study was to examine the impact of removable magnets on biofilm formation in an in vitro model. METHODS: Silastic models were constructed to represent CIs with and without a magnet pocket and with and without a titanium blank in the pocket. CIs were exposed to a culture of a biofilm forming strain of Staphylococcus aureus. Adherence of planktonic bacteria and biofilm formation were assessed with quantitative bacterial counts and scanning electron microscopy. RESULTS: Adherent bacterial counts were significantly higher in CI models with an empty magnet pocket (P = 0.0097). Biofilm formation was significantly lower in CI models without a magnet pocket (P = 0.0121). CONCLUSIONS: CI magnet pockets harbor bacteria that can increase biofilm development in an in vitro model. PMID- 17418257 TI - Threshold shift: effects of cochlear implantation on the risk of pneumococcal meningitis. AB - OBJECTIVES: The study goals were to examine whether cochlear implantation increases the risk of meningitis in the absence of other risk factors and to understand the pathogenesis of pneumococcal meningitis post cochlear implantation. STUDY DESIGN AND SETTING: Four weeks following surgery, 54 rats (18 of which received a cochleostomy alone, 18 of which received a cochleostomy and acute cochlear implantation using standard surgical techniques, and 18 of which received a cochlear implant) were infected with Streptococcus pneumoniae via three different routes of bacterial inoculation (middle ear, inner ear, and intraperitoneal) to represent all potential routes of bacterial infection from the upper respiratory tract to the meninges. RESULTS: The presence of a cochlear implant reduced the threshold of bacteria required to cause pneumococcal meningitis from all routes of infection in healthy animals. CONCLUSION: The presence of a cochlear implant increases the risk of pneumococcal meningitis regardless of the route of bacterial infection. SIGNIFICANCE: Early detection and treatment of pneumococcal infection such as otitis media may be required, as cochlear implantation may lead to a reduction of infectious threshold for meningitis. PMID- 17418258 TI - Cochlear implantation and quality of life in postlingually deaf adults: long-term follow-up. AB - OBJECTIVE: To investigate long-term quality of life (QoL) in postlingually deaf adults after entering the cochlear implantation (CI) program. STUDY DESIGN AND SETTING: Follow-up study from 1998 onwards in tertiary university medical center. Long-term CI users, patients who have not received a CI, and relatively short term CI users were re-evaluated six years after initial data collection in 1998 by using three questionnaires (NCIQ, HUI3, and SF36) and speech perception tests. RESULTS AND CONCLUSIONS: In general, the beneficial effect of CI remained stable during long-term follow-up, though scores on the questionnaires decreased slightly. Outcomes before and after cochlear implantation were significantly different. The group without a CI demonstrated slightly decreasing trends in outcomes. Long-term speech perception performance improved in time. SIGNIFICANCE: This is the first study to investigate long-term follow-up of CI patients, in all aspects of QoL combined with speech perception performance, in comparison with postlingually deaf adults without CI. PMID- 17418259 TI - Intranasal corticosteroid use is associated with lower rates of bacterial recovery in chronic rhinosinusitis. AB - OBJECTIVE: To determine whether use of a topical intranasal corticosteroid (INCS) preoperatively had an effect on the bacterial recovery rate and flora recovered at endoscopic sinus surgery (ESS). STUDY DESIGN AND SETTING: A prospective, multicenter, observational study from academic-based rhinology practices. Consecutive, unselected patients undergoing ESS had protected sinus cultures done at the time of ESS. RESULTS: 157 patients were assessed. Overall growth rate was 45.5%. INCS users had a positive culture rate of 35.4% vs 61.7% in nonusers (P = 0.0001). This effect was most pronounced in the subgroup undergoing revision surgeries (bacterial recovery rate INCS: 40.0%, no INCS: 82.6%, P = 0.001) and most marked for S. aureus (INCS: 12.5%, no INCS: 40.0%, P = 0.04) and CNS (INCS: 12.5%, no INCS: 30.4%, P = 0.05). While the rate of nasal polyposis was higher in both revision and the INCS-treated groups, rate of bacterial recovery was not influenced by a diagnosis of nasal polyposis. CONCLUSION: INCS use preoperatively is associated with a lesser rate of bacterial recovery at the time of ESS, especially in individuals with previous ESS. SIGNIFICANCE: The results suggest a possible role of INCS in the management of post-ESS disease. PMID- 17418260 TI - The clinical behavior of isolated sphenoid sinusitis. AB - OBJECTIVE: We sought to study the clinical behavior and treatment outcome of isolated sphenoid sinusitis (ISS). STUDY DESIGN AND SETTING: We conducted a retrospective study of patients diagnosed with ISS in a tertiary medical center over 20 years. RESULTS: Of 72 patients with ISS, 79 percent had acute symptoms, 15 percent had chronic symptoms, and 6 percent had incidental radiological findings. Fifteen percent were children. Most patients were diagnosed between January and April (P < 0.01), and increasing incidence was noted over the years (P < 0.001). Headache was the most common presenting symptom (85%). Chronic patients complained also of nasal symptoms (82%). Six patients had a major complication of sinusitis (none of them were children), and two patients died. Immunocompromised patients had more major complications (P or =70% (50% vs. 35%; p = 0.03) and MBG 2 or 3 (79% vs. 58%; p = 0.001). Left ventricular function recovery at 1 month was significantly greater in the early group (mean gain ejection fraction 8 +/- 7% vs. 6 +/- 7%, p = 0.02; mean gain wall motion score index 0.4 +/- 0.3 vs. 0.3 +/- 0.3, p = 0.03). CONCLUSIONS: In patients with AMI treated with primary PCI, early abciximab administration improves pre-PCI angiographic findings, post-PCI tissue perfusion, and 1-month left ventricular function recovery, possibly by starting early recanalization of the infarct related artery. PMID- 17418290 TI - Prognostic value of midregional pro-adrenomedullin in patients with acute myocardial infarction: the LAMP (Leicester Acute Myocardial Infarction Peptide) study. AB - OBJECTIVES: This study sought to assess the prognostic impact of midregional pro adrenomedullin (MR-proADM) after an acute myocardial infarction (AMI). BACKGROUND: Adrenomedullin (ADM) is elevated in heart failure (HF) and after AMI. Another part of its precursor, MR-proADM, is more stable in circulation and ex vivo. We investigated the cardiovascular prognostic value after AMI of MR-proADM and compared it with N-terminal pro-B-type natriuretic peptide (NTproBNP), a marker of death and HF. METHODS: We measured plasma MR-proADM and NTproBNP in 983 consecutive post-AMI patients (721 men, mean age 65.0 +/- 12.2 years), 3 to 5 days after chest pain onset. RESULTS: There were 101 deaths and 49 readmissions with HF during follow-up (median 342, range 0 to 764 days). The MR-proADM was increased in patients with death or HF compared with survivors (median 1.19 nmol/l, range 0.09 to 5.39 nmol/l, vs. 0.71 nmol/l, range 0.25 to 6.66 nmol/l, p < 0.0001). Using a multivariate binary logistic model, log MR-proADM (odds ratio 4.22) and log NTproBNP (odds ratio 3.20) were significant independent predictors of death or HF (with creatinine, age, gender, and history of AMI). The areas under the receiver-operating characteristic curve for MR-proADM, NTproBNP, and the logistic model with both markers were 0.77, 0.79, and 0.84 respectively. Cox models for the predictors of death or HF showed the same variables (including log MR-proADM, hazard ratio 3.63; log NTproBNP, hazard ratio 2.67). The MR-proADM provided further risk stratification in those patients who had NTproBNP levels above the median (p < 0.0001). Findings were similar for death and HF as individual end points. CONCLUSIONS: The ADM system is activated after AMI. The MR proADM is a powerful predictor of adverse outcome, especially in those with an elevated NTproBNP. The MR-proADM may represent a clinically useful marker of prognosis after AMI. PMID- 17418291 TI - Vascular inflammation evaluated by [18F]-fluorodeoxyglucose positron emission tomography is associated with the metabolic syndrome. AB - OBJECTIVES: We investigated factors for carotid artery inflammation by [(18)F] fluorodeoxyglucose (FDG) positron emission tomography (PET). BACKGROUND: Inflammation is present in some atherosclerotic plaques. The FDG-PET is capable of identifying and quantifying vascular inflammation within atherosclerotic plaques. METHODS: The FDG-PET imaging was performed in 216 consecutive patients (63 +/- 9 years, men:women 147:69) for cancer screening. Vascular inflammation in carotid atherosclerosis was quantified by measuring the standardized uptake value (SUV) of FDG into the artery. RESULTS: Multiple stepwise regression analysis revealed significant relationships between SUV and waist circumference (p < 0.001), hypertensive medication (p < 0.001), carotid intima-media thickness (p < 0.001), high-density lipoprotein cholesterol (p < 0.01, inversely), homeostasis model assessment of insulin resistance (p < 0.05), or high sensitivity C-reactive protein (p < 0.05). Age- and gender-adjusted SUV of FDG was significantly higher (p < 0.0001) in proportion to the accumulation of the number of the components of the metabolic syndrome. Thus, the metabolic syndrome was associated with increased FDG uptake in carotid atherosclerosis. CONCLUSIONS: Our present study may suggest that the metabolic syndrome is associated with inflammation in carotid atherosclerosis. (Detection of Plaque Inflammation by Positron Emission Tomography (PET); http://www.clinicaltrials.gov/ct/show/NCT00114504; NCT00114504). PMID- 17418292 TI - The vital prognosis of subclavian stenosis. AB - OBJECTIVES: This study sought to assess the prognosis of subclavian stenosis (SS) as a potential marker of total and cardiovascular disease (CVD) mortality. BACKGROUND: Subclavian stenosis, diagnosed by a brachial systolic pressure difference (BSPD) > or =15 mm Hg, is associated with an increased prevalence of CVD risk factors. However, the association between SS and mortality is unknown. We hypothesized that a BSPD > or =15 mm Hg would predict an increased risk of CVD events. METHODS: We analyzed baseline and longitudinal data from 3 cohorts. Two were recruited from noninvasive vascular laboratories, and the third was a community-dwelling cohort. Multivariate survival models were used to test for an independent association of SS with total and CVD mortality. RESULTS: Baseline and follow-up data (mean 9.8 years) were complete in 1,778 participants. Subclavian stenosis was found in 157 (8.8%) subjects. Adjusted for age, gender, ethnicity, and cohort of origin, the presence of SS was significantly associated with increased total and CVD mortality (respectively, hazard ratio [HR] 1.42, p < 0.005; and HR 1.50, p = 0.05). This association persisted after adjustments for CVD risk factors (smoking pack-years, hypertension, diabetes, total/high-density lipoprotein cholesterol ratio, and body mass index) as well as lipid-lowering and antiplatelet therapies (HR 1.40, p < 0.01; and HR 1.57, p < 0.05 for total and CVD mortality, respectively). When any history of CVD or an ankle-brachial index <0.90 were added to the model, SS remained an independent predictor for total mortality (HR 1.34, p = 0.02), with a similar trend for CVD mortality (HR 1.43, p = 0.09). CONCLUSIONS: The presence of SS, easily diagnosed by comparing systolic pressures in the left and right arm, predicts total and CVD mortality independent of both CVD risk factors and existent CVD at baseline. PMID- 17418293 TI - Rate of progression of coronary atherosclerotic plaque in women. AB - OBJECTIVES: The purpose of this study was to determine the relationship between gender and the extent of coronary atherosclerosis assessed by intravascular ultrasound (IVUS) and its rate of progression in subjects treated with established medical therapies. BACKGROUND: It is uncertain whether the pathophysiology of coronary artery disease (CAD) differs between genders. METHODS: A systematic analysis was performed of 978 subjects who participated in serial studies of atheroma progression. Genders were compared with regard to the extent of coronary atheroma at baseline and subsequent change in response to use of established medical therapies. RESULTS: Women were more likely to have a history of hypertension and higher levels of body mass index, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, C reactive protein, and systolic and diastolic blood pressure. Despite this, women had less plaque in terms of percent atheroma volume (PAV) (33.9 +/- 10.2% vs. 37.8 +/- 10.3%, p < 0.001) and total atheroma volume (TAV) (148.7 +/- 66.6 mm3 vs. 194.7 +/- 84.3 mm3, p < 0.001). With medical therapy, the rate of change of PAV (0.7 +/- 0.6% vs. 0.7 +/- 0.5%, p = 0.92) and TAV (-2.3 +/- 3.2 mm3 vs. -1.9 +/- 2.9 mm3, p = 0.84) did not differ between genders. In the setting of intensive risk factor modification, there was no significant difference between genders with regard to the rates of plaque progression or regression. CONCLUSIONS: Despite the presence of more risk factors, the extent of atheroma in women with angiographic CAD is less than in men in subjects participating in clinical trials that employed serial assessments with IVUS. The finding that the rate of plaque progression or regression does not differ between genders in the setting of intensive risk factor modification supports the use of established medical therapies in women with CAD. PMID- 17418294 TI - Significant improvement in short-term mortality in women undergoing coronary artery bypass surgery (1991 to 2004). AB - OBJECTIVES: This study sought to evaluate gender differences and trends in 30-day mortality after coronary artery bypass surgery (CABG). BACKGROUND: Evidence for gender differences in short-term mortality after CABG is conflicting. Many studies were from single centers, included highly selected populations, or had limited clinical information for adequate covariate adjustment. We undertook a population-based analysis using detailed clinical data on all adults undergoing CABG in the province of British Columbia, Canada. METHODS: The study population comprised all residents 20 years and older who underwent isolated CABG between 1991 and 2004. Multiple logistic regression was used to examine the association between gender and 30-day mortality; time trend analysis was conducted by Mantel Haenszel chi-square test. RESULTS: The study cohort comprised 20,229 men and 4,983 women. Women were older and had more comorbid conditions than men, but had better ejection fractions and less extensive coronary disease. Thirty-day mortality decreased significantly in men (2.4% to 1.9%) and women (5.6% to 1.9%) over the 14-year study period. Overall, 30-day mortality was significantly higher in women (3.6% vs. 2.0%, p < 0.001), and adjustment for baseline differences did not remove this difference (odds ratio 1.42, 95% confidence interval 1.15 to 1.75). Adjustment for body surface area, an intrinsic gender difference, further attenuated the relationship (odds ratio 1.26, 95% confidence interval 0.96 to 1.64). CONCLUSIONS: The 30-day mortality after CABG decreased significantly between 1991 and 2004, especially in women, suggesting that the gender difference in short-term outcomes is diminishing. The overall 42% higher mortality risk in women seems to be partially mediated through body surface area, a surrogate for vessel size. PMID- 17418295 TI - The controversy in clinical results among men and women after coronary bypass operation. PMID- 17418296 TI - Trends in anticoagulation for atrial fibrillation in the U.S.: an analysis of the national ambulatory medical care survey database. AB - OBJECTIVES: The purpose of this study was to track trends in the use of anticoagulation for atrial fibrillation (AF) over the past decade and identify predictors of use. BACKGROUND: Atrial fibrillation is common and associated with significant morbidity. Previous studies suggest underuse of anticoagulant therapy in patients with AF. METHODS: The National Ambulatory Medical Care Survey database was queried for all patient visits with a diagnosis of AF between 1994 and 2003. Other diagnoses, other medications, and demographic, visit, geographic, and provider characteristics were compared with the prescription of anticoagulation in predefined age and risk groups. RESULTS: The prevalence of the diagnosis of AF and anticoagulation for AF has increased over the last decade. Increased age and use of rate control agents is associated with the use of anticoagulation. There is a trend toward less anticoagulation when a rhythm control agent is used instead of a rate control agent. Anticoagulation might be overused in a group of low-risk patients. CONCLUSIONS: From 1995 through 2002, an increase has occurred in anticoagulation for AF, especially in those at highest risk for thromboembolic phenomena. A substantial number of patients at risk for thromboembolic events are not anticoagulated, and further studies are needed to determine how many of these patients are candidates for anticoagulation. Anticoagulation use has increased in nontargeted, low-risk groups in whom antiplatelet agents are appropriate. Use of a rhythm control agent might be associated with less use of anticoagulation. PMID- 17418297 TI - Transplantation of autologous endothelial progenitor cells may be beneficial in patients with idiopathic pulmonary arterial hypertension: a pilot randomized controlled trial. AB - OBJECTIVES: The goal of this study was to investigate the feasibility, safety, and initial clinical outcome of intravenous infusion of autologous endothelial progenitor cells (EPCs) in patients with idiopathic pulmonary arterial hypertension (IPAH). BACKGROUND: Experimental data suggest that transplantation of EPCs attenuates monocrotaline-induced pulmonary hypertension in rats and dogs. In addition, clinical studies suggest that autologous progenitor cell transplantation is feasible and safe in patients with ischemic diseases. METHODS: We conducted a prospective, randomized trial comparing the effects of EPC transplantation plus conventional therapy with those of conventional therapy alone in patients with IPAH. The primary end point was change in the 6-min walk distance using a standardized protocol. The secondary end points were changes in hemodynamic variables as assessed by right heart catheterization. RESULTS: After 12 weeks of follow-up, the mean distance walked in 6 min increased by 48.2 m in the cell infusion group (from 263 +/- 42 m to 312 +/- 34 m), and an increase of 5.7 m occurred in the conventional therapy group (from 264 +/- 42 m to 270 +/- 44 m). The mean difference between the 2 groups was 42.5 m (95% confidence interval 28.7 to 56.3 m, p < 0.001). The patients in the cell infusion group also had significant improvement in mean pulmonary artery pressure, pulmonary vascular resistance, and cardiac output. There were no severe adverse events with cell infusion. CONCLUSIONS: This preliminary study showed that intravenous infusion of autologous EPCs seemed to be feasible and safe, and might have beneficial effects on exercise capacity and pulmonary hemodynamics in patients with IPAH. (Safety and Efficacy Study of Transplantation of EPCs to Treat Idiopathic Pulmonary Arterial Hypertension; http://www.clinicaltrials.gov/ct/show/NCT00257413?order=1; NCT00257413). PMID- 17418298 TI - Is it possible to reverse the endothelial dysfunction in pulmonary arterial hypertension? PMID- 17418299 TI - Endothelial nitric oxide synthase overexpression provides a functionally relevant angiogenic switch in hibernating pig myocardium. AB - OBJECTIVES: We investigated whether retroinfusion of liposomal endothelial nitric oxide synthase (eNOS) S1177D complementary deoxyribonucleic acid (cDNA) would affect neovascularization and function of the ischemic myocardium. BACKGROUND: Recently, we demonstrated the feasibility of liposomal eNOS cDNA transfection via retroinfusion in a model of acute myocardial ischemia/reperfusion. In the present study, we used this approach to target a phosphomimetic eNOS construct (eNOS S1177D) into chronic ischemic myocardium in a pig model of hibernation. METHODS: Pigs (n = 6/group) were subjected to percutaneous implantation of a reduction stent graft into the left anterior descending artery (LAD), inducing total occlusion within 28 days. At day 28, retroinfusion of saline solution containing liposomal green fluorescent protein or eNOS S1177D cDNA (1.5 mg/animal, 2 x 10 min) was performed. Furthermore, L-nitroarginine-methylester (L-NAME) was applied orally from day 28, where indicated. At day 28 and day 49, fluorescent microspheres were injected into the left atrium for perfusion analysis. Regional functional reserve (at atrial pacing 140/min) was assessed at day 49 by subendocardial segment shortening (SES) (sonomicrometry, percent of ramus circumflexus region). RESULTS: The eNOS S1177D overexpression increased endothelial cell proliferation as well as capillary and collateral growth at day 49. Concomitantly, eNOS S1177D overexpression enhanced regional myocardial perfusion from 62 +/- 4% (control) to 77 +/- 3% of circumflex coronary artery perfused myocardium, unless L-NAME was co-applied (69 +/- 5%). Similarly, eNOS S1177D cDNA improved functional reserve of the LAD (33 +/- 5% vs. 7 +/- 3% of circumflex coronary artery-perfused myocardium), except for L-NAME coapplication (13 +/- 6%). CONCLUSIONS: Retroinfusion of eNOS S1177D cDNA induces neovascularization via endothelial cell proliferation and collateral growth. The resulting gain of perfusion enables an improved functional reserve of the hibernating myocardium. PMID- 17418301 TI - Endothelial progenitor cells in coronary artery disease. PMID- 17418303 TI - Adjunctive thrombectomy in acute myocardial infarction: for some but not for all. PMID- 17418304 TI - PCR examination of bronchoalveolar lavage samples is a useful tool in pre clinical diagnosis of ovine pulmonary adenocarcinoma (Jaagsiekte). AB - Ovine pulmonary adenocarcinoma (OPA) is a contagious lung tumour of sheep caused by Jaagsiekte sheep retrovirus (JSRV). The disease is a particular problem in flocks in many parts of the world. The aim of the study was to assess screening methods for individual animals as a prelude to future eradication trials. Results of histological examination were used as the standard to evaluate the relative sensitivity and specificity of an established heminested polymerase chain reaction (PCR) test for JSRV proviral DNA from blood and bronchoalveolar lavage (BAL) samples. PCR results from tissue samples are included as control data. PCR testing of blood samples was found to have an estimated sensitivity of only 10% (95% confidence interval (CI) 3-20) while the sensitivity of the PCR test on BAL samples was 89% (CI 79-96) in comparison to the results of histological examination. We conclude that PCR testing of BAL samples is an effective confirmatory test for sheep with suspected clinical OPA. It is also a useful tool for the pre-clinical identification of individual infected sheep within an infected flock and therefore may prove beneficial in future control or eradication programmes. PMID- 17418305 TI - Vibrio parahaemolyticus: a concern of seafood safety. AB - Vibrio parahaemolyticus is a human pathogen that is widely distributed in the marine environments. This organism is frequently isolated from a variety of raw seafoods, particularly shellfish. Consumption of raw or undercooked seafood contaminated with V. parahaemolyticus may lead to development of acute gastroenteritis characterized by diarrhea, headache, vomiting, nausea, and abdominal cramps. This pathogen is a common cause of foodborne illnesses in many Asian countries, including China, Japan and Taiwan, and is recognized as the leading cause of human gastroenteritis associated with seafood consumption in the United States. This review gives an overview of V. parahaemolyticus food poisoning and provides information on recent development in methods for detecting V. parahaemolyticus and strategies for reducing risk of V. parahaemolyticus infections associated with seafood consumption. PMID- 17418306 TI - Antibiotic resistance in non-enterococcal lactic acid bacteria and bifidobacteria. AB - Over the last 50 years, human life expectancy and quality of life have increased dramatically due to improvements in nutrition and the use of antibiotics in the fight against infectious diseases. However, the heyday of antibiotic treatment is on the wane due to the appearance and spread of resistance among harmful microorganisms. At present, there is great concern that commensal bacterial populations from food and the gastrointestinal tract (GIT) of humans and animals, such as lactic acid bacteria (LAB) and bifidobacteria, could act as a reservoir for antibiotic resistance genes. Resistances could ultimately be transferred to human pathogenic and opportunistic bacteria hampering the treatment of infections. LAB species have traditionally been used as starter cultures in the production of fermented feed and foodstuffs. Further, LAB and bifidobacteria are normal inhabitants of the GIT where they are known to exert health-promoting effects, and selected strains are currently been used as probiotics. Antibiotic resistance genes carried by LAB and bifidobacteria can be transferred to human pathogenic bacteria either during food manufacture or during passage through the GIT. The aim of this review is to address well-stated and recent knowledge on antibiotic resistance in typical LAB and bifidobacteria species. Therefore, the commonest antibiotic resistance profiles, the distinction between intrinsic and atypical resistances, and some of the genetic determinants already discovered will all be discussed. PMID- 17418307 TI - Significance of yeasts in the fermentation of maize for ogi production. AB - Saccharomyces cerevisiae, Candida krusei, C. tropicalis, Geotrichum candidum, G. fermentans and Rhodotorula graminis were isolated during the fermentation of maize for ogi production. All the isolates except Geotrichum fermentans and Rhodotorula graminis were able to degrade phytate. All the yeasts strains exhibited lipase and esterase activities. Only S. cerevisiae (2.60%) and C. krusei (7.41%) exhibited amylase activities. Candida sp. produced wider zone of inhibition than the other yeasts strains tested during lipase activity while S. cerevisiae strains produced significantly wider zone of clearing as compared to the other yeasts for esterase activities. The study of inter-relationships between Lactobacillus plantarum and yeasts (C. krusei and S. cerevisiae) showed that the growth of the yeast strains were enhanced during fermentation by the presence of the lactic acid bacteria, but the growth of the L. plantarum strain was significantly enhanced especially by the C. krusei. PMID- 17418308 TI - Dynamics and characterization of yeasts during ripening of typical Italian dry cured ham. AB - The evolution of the yeast population during manufacturing and ripening of dry cured Parma ham was investigated. Contamination levels ranged from 10(5) to 10(7) cfu/g on muscle surface, 10(4) to 10(6) cfu/g on covering fat and exceeded 10(7) cfu/g on spreadable fat mince ("sugna"). Two hundred and sixty one yeast isolates underwent identification test, showing that the predominant species of yeast population during the whole maturing process were Debaryomyces hansenii, Candida zeylanoides, Debaryomyces maramus, and to a lesser extent, Candida famata and Hyphopichia burtonii. The species Candida catenulata, Candida guilliermondii, Candida edax and other genera like Cryptococcus and Wingea were occasionally found. The yeast counts and species distribution changed according to the stage of processing and to the ham sampling location. At the end of the cold phase, the washing procedure was effective in lowering the yeast count in muscle and fat surface layers, but during the next ageing stages, yeast colonization of unskinned ham muscle increased again, though species distribution changed if compared to previous manufacturing phases. The ripening steps taken into account from the end of the cold phase to the final outcome, were always characterized by more than one yeast species, suggesting that yeasts other than Debaryomyces spp. could play a remarkable role on the sensory and safety properties of typical Italian dry-cured ham. PMID- 17418309 TI - Adhesion and detachment kinetics of several strains of Staphylococcus aureus subsp. aureus under three different experimental conditions. AB - The kinetics of adhesion of five Staphylococcus aureus subsp. aureus strains (CECT 976, 4459, 4465, 4466 and 5191) to polypropylene at 25 degrees C in the absence of nutrients (PBS medium) were initially compared. Those strains with the highest (CECT 4459) and the lowest (CECT 976) adhesion levels were selected for further studying the effects of a nutrient-rich adhesion-promoting medium (TSB plus 1% glucose-TSBG) as well as of a conditioning film consisting of dried mussel cooking juices (MCJ) on adhesion to and detachment from polypropylene surfaces. Adhesion kinetics were properly described by an empirical model in the absence of conditioning film. The maximum adhesion level was much higher in the presence of TSBG than in PBS, decreasing sharply in both cases after 10-15 h. In contrast, adhesion increased exponentially during 25 h in the presence of dried MCJ. Clear differences were thus found in different media, and it suggests that cleaning strategies should vary under different conditions. The comparison of the adhesion strengths under the different experimental conditions showed that the persistence was highest when biofilms were formed on MCJ, which indicates that cells would remain longer as a source of cross-contamination. Some biofilms were examined by electronic microscopy, and different structures were observed under the different experimental conditions. It is concluded that the study of biofilm formation by S. aureus is necessary to establish efficient control systems in the food industry. PMID- 17418310 TI - Identification of subdominant sourdough lactic acid bacteria and their evolution during laboratory-scale fermentations. AB - Presumptive lactic acid bacterial cocci were found in six sourdoughs (out of 20) from the Abruzzo region (central Italy) and subjected to phenotypic and genotypic characterization. A total of 21 isolates, recognized as seven strains by randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing, were identified by a polyphasic approach, consisting of 16S rRNA gene sequencing, multiplex PCR assays and physiological features, as Enterococcus faecium and Pediococcus pentosaceus. Four strains belonging to those species and previously isolated from wheat kernels were inoculated in sterile flour to verify their capacity to grow in sourdough environment. Doughs with several dual bacterial combinations, including Lactobacillus sanfranciscensis, were propagated for 11 days and pH measurements and bacterial counts were carried out. PMID- 17418311 TI - Selection and properties of alpha-acetolactate decarboxylase-deficient spontaneous mutants of Streptococcus thermophilus. AB - Many lactic acid bacteria produce diacetyl, which is a desirable aroma compound in some fermented dairy products. Strains or mutants of Lactococcus lactis subsp. lactis biovar. diacetylactis that are deficient in alpha-acetolactate decarboxylase are used in some food processes for their ability to produce large amounts of diacetyl. However, up until now, the use of alpha-acetolactate decarboxylase-deficient mutants of Streptococcus thermophilus for increased diacetyl production has not been evaluated. The objective of the present study was to devise a procedure for selecting spontaneous alpha-acetolactate decarboxylase-deficient mutants of S. thermophilus. We observed that in a chemically defined medium (CDM) containing alpha-ketobutyrate plus leucine, or alpha-ketobutyrate plus leucine plus isoleucine, the alpha-acetolactate decarboxylase-deficient mutant TIL865, obtained by directed mutagenesis, grew faster than its parent strain. This property was used for selecting spontaneous alpha-acetolactate decarboxylase-deficient mutants on agar plates. The resulting mutants were able to grow in milk, and their acidifying activity was slightly lower than that of the parent strain. Under partial anaerobic or aerobic conditions, they produced approximately three times more diacetyl than the parent strain. Such spontaneous mutants may be useful for increasing the diacetyl content of fermented milks whose production involves S. thermophilus strains. PMID- 17418312 TI - Combined effect of oregano essential oil and modified atmosphere packaging on shelf-life extension of fresh chicken breast meat, stored at 4 degrees C. AB - The combined effect of oregano essential oil (0.1% and 1% w/w) and modified atmosphere packaging (MAP) (30% CO2/70% N2 and 70% CO2/30% N2) on shelf-life extension of fresh chicken meat stored at 4 degrees C was investigated. The parameters that were monitored were: microbiological (TVC, Pseudomonas spp., lactic acid bacteria (LAB), yeasts, Brochothrix thermosphacta and Enterobacteriaceae), physico-chemical (pH, TBA, color) and sensory (odor and taste) attributes. Microbial populations were reduced by 1-5 log cfu/g for a given sampling day, with the more pronounced effect being achieved by the combination of MAP and oregano essential oil. TBA values for all treatments remained lower than 1 mg malondialdehyde (MDA) kg(-1) throughout the 25-day storage period. pH values varied between 6.4 (day 0) and 5.9 (day 25). The values of the color parameters L*, a* and b* were not considerably affected by oregano oil or by MAP. Finally, sensory analysis showed that oregano oil at a concentration of 1% imparted a very strong taste to the product for which reason these lots of samples were not scored. On the basis of sensory evaluation a shelf life extension of breast chicken meat by ca. 3-4 days for samples containing 0.1% oregano oil, 2-3 days for samples under MAP and 5-6 days for samples under MAP containing 0.1% of oregano oil was attained. Thus oregano oil and MAP exhibited an additive preservation effect. PMID- 17418313 TI - Chemical and microbiological characterisation of "Salpicao de Vinhais" and "Chourica de Vinhais": traditional dry sausages produced in the North of Portugal. AB - "Chourica de Vinhais" and "Salpicao de Vinhais" are traditional smoked naturally fermented meat products produced in the North of Portugal, Tras-os-Montes. The objective of this study was the characterisation of these products, giving particular attention to their microbiological and chemical safety. Nitrite, nitrate, heavy metals and biogenic amines were within accepted limits for meat products. Globally, the need for improvements in the good manufacturing practices was demonstrated in this study as various lots were considered of unsatisfactory microbiological quality according to the guidelines published by the Food Safety Authority of Ireland. PMID- 17418314 TI - Response surface model for prediction of growth parameters from spores of Clostridium sporogenes under different experimental conditions. AB - Clostridium sporogenes is considered to be a non-toxingenic equivalent of proteolytic Clostridium botulinum, and it also causes food spoilage. The effects of temperature (16.6-33.4 degrees C), pH value (5.2-6.8) and concentration of sodium chloride (0.6-7.4%) on the growth parameters of C. sporogenes spores were investigated. The growth curves generated within different conditions were fitted using Baranyi function. Two growth parameters (growth rate, GR; lag-time, LT) of the growth curves under combined effects of temperature, pH and sodium chloride were modeled using a quadratic polynomial equation of response surface (RS) model. Mathematical evaluation demonstrated that the standard error of prediction (%SEP) obtained by RS model was 1.033% for GR and was 0.166% for LT for model establishing. The %SEP for model validation were 43.717% and 5.895% for GR and LT, respectively. The root-mean-squares error (RMSE) was in acceptable range which was less than 0.1 for GR and was less than 8.0 for LT. Both the bias factor (B(f)) and accuracy factor (A(f)) approached 1.0, which were within acceptable range. Therefore, RS model provides a useful and accurate method for predicting the growth parameters of C. sporogenes spores, and could be applied to ensure food safety with respect to proteolytic C. botulinum control. PMID- 17418315 TI - Sensitivity of two techniques to detect Escherichia coli O157 in naturally infected bovine fecal samples. AB - The aim of this study was to investigate the sensitivity and reliability of two techniques commonly used for the isolation of Escherichia coli O157: (i) buffered peptone water (BPW) containing vancomycin, cefsulodin and cefixime followed by immunomagnetic separation (IMS-VCC) and (ii) modified E. coli (EC) broth supplemented with novobiocin (m ECn), both followed by culturing on cefixime tellurite sorbitol McConkey (ctSMAC) agar plates. Over a 2-year period, 24 feedlots located over a large geographical area (approximately 600 x 450 km) were screened for the presence of E. coli O157. A total of 194 E. coli O157 isolates were identified; 151 (77.4%) using IMS-VCC and 108 (55.4%) using m ECn. The recovery rates of IMS-VCC varied from 100% to 47%, whereas for m ECn ranged from 100% to 16%. All isolates were grouped, using randomly amplified polymorphic DNA polymerase chain reaction (RAPD-PCR), into 3 major clusters that comprised 39 different subtypes, providing evidence of considerable genetic heterogeneity. The results from this study revealed false negatives in IMS-VCC technique, most probably due to the high genetic diversity of environmental E. coli O157 isolates and antibiotic sensitivity. Using only IMS-VCC as a method for detection may result in significant underestimation of the pathogen. Performing two different enrichment steps in parallel can lead to markedly improved recovery rates of E. coli O157 isolates from naturally infected samples. PMID- 17418316 TI - Predictive models for growth of Salmonella typhimurium DT104 from low and high initial density on ground chicken with a natural microflora. AB - A single strain (ATCC 700408) of Salmonella typhimurium DT104 was used to investigate and model growth from a low (1.12 log10 mpn g(-1)) and high (3.7 log10 cfu g(-1)) initial density on ground chicken with a natural microflora. Kinetic data for growth of the pathogen on ground chicken were fit to a primary model to determine lag time (lambda), maximum specific growth rate (mu) and maximum population density (Nmax). Secondary models for lambda, mu and Nmax, as a function of temperature (10-40 degrees C), were developed and compared among initial densities. Variation of pathogen growth among replicates (n=4 or 5) was higher at 10-18 degrees C than at 22-40 degrees C and was higher for Nmax than lambda and mu. Prediction problems were observed when secondary models developed with one initial density were used to predict lambda, mu and Nmax from the other initial density, especially at 10-18 degrees C and for Nmax. These results indicated that variation of growth among replicate challenge studies and initial density are important factors to consider when developing predictive models for growth of S. typhimurium DT104 on ground chicken with a natural microflora. PMID- 17418317 TI - Measurement uncertainty of the EU methods for microbiological examination of red meat. AB - Three parallel trials were made of EU methods proposed for the microbiological examination of red meat using two analysts in each of seven laboratories within the UK. The methods involved determination of aerobic colony count (ACC) and Enterobacteriaceae colony count (ECC) using simulated methods and a freeze-dried standardised culture preparation. Trial A was based on a simulated swab test, Trial B a simulated meat excision test and Trial C was a reference test on reconstituted inoculum. Statistical analysis (ANOVA) was carried out before and after rejection of outlying data. Expanded uncertainty values (relative standard deviation x2) for repeatability and reproducibility, based on the log10 cfu/ml, on the ACC ranged from +/-2.1% to +/-2.7% and from +/-5.5% to +/-10.5%, respectively, depending upon the test procedure. Similarly for the ECC, expanded uncertainty estimates for repeatability and reproducibility ranged from +/-4.6% to +/-16.9% and from +/-21.6% to +/-23.5%, respectively. The results are discussed in relation to the potential application of the methods. PMID- 17418318 TI - Ethylene inhibits aflatoxin biosynthesis in Aspergillus parasiticus grown on peanuts. AB - The filamentous fungi Aspergillus parasiticus and Aspergillus flavus synthesize aflatoxins when they grow on a variety of susceptible food and feed crops. These mycotoxins are among the most carcinogenic naturally occurring compounds known and they pose significant health risks to humans and animals. We previously demonstrated that ethylene and CO2 act alone and together to reduce aflatoxin synthesis by A. parasiticus grown on laboratory media. To demonstrate the potential efficacy of treatment of stored seeds and grains with these gases, we tested ethylene and CO2 for ability to inhibit aflatoxin accumulation on Georgia Green peanuts stored for up to 5 days. We demonstrated an inverse relationship between A. parasiticus spore inoculum size and the level of toxin accumulation. We showed that ethylene inhibits aflatoxin synthesis in a dose-dependent manner on peanuts; CO2 also inhibits aflatoxin synthesis over a narrow dose range. Treatments had no discernable effect on mold growth. These observations support further exploration of this technology to reduce aflatoxin contamination of susceptible crops in the field and during storage. PMID- 17418319 TI - Recovery and detection of Vibrio vulnificus during cold storage. AB - Different cultural techniques and molecular methods for the detection of Vibrio vulnificus during cold storage in a model broth system were compared. Two strains of V. vulnificus were grown to stationary phase and inoculated (10(6) CFU/mL) into tryptic soy broth with 2% sodium chloride (TSBN2) or artificial seawater (ASW), both pre-chilled to 5 degrees C. These were stored for 10 days, with sub sampling conducted at time 0 and every 2 days thereafter. Each subsample was plated, by both pour and spread plate techniques, onto tryptic soy agar 2% sodium chloride (TSAN2) with or without catalase (400 or 600 U) or sodium pyruvate (80 or 160 mg) supplementation. Nucleic acids were extracted from subsamples and subjected to PCR and RT-PCR with hemolysin as the target. Higher recoveries of V. vulnificus were obtained with spread plating compared to pour plating (P<0.05). The addition of sodium pyruvate (80 mg) or catalase (400 U) significantly increased cell recovery (P<0.05). PCR amplification signals were stronger than RT PCR signals at each timepoint, and results were generally consistent between TSAN2 and ASW for each strain. These results will aid in the design of optimum methods to recover and/or detect V. vulnificus cells subjected to sublethal stress that might be encountered in food processing and storage. PMID- 17418320 TI - Confirmation of dengue virus infection by detection of dengue virus type 1 genome in urine and saliva but not in plasma. AB - We successfully detected dengue virus (DENV) genome in urine and saliva but not in plasma samples from a Japanese dengue fever patient. The results of the present study suggest that detection of DENV genome in urine and saliva can be an effective diagnostic method, particularly for children with viral hemorrhage. PMID- 17418321 TI - Effect of hygiene promotion on the risk of reinfection rate of intestinal parasites in children in rural Uzbekistan. AB - A controlled prospective cohort study was designed to assess the effectiveness of hygiene promotion on the risk of reinfection by intestinal parasites in children in 276 rural Uzbek households over a 1-year period. The study included three groups: seasonality (no medicine, no hygiene promotion activity), treatment (medicine provided, no hygiene promotion activity) and hygiene promotion (medicine provided and hygiene promotion activity). The Participatory Hygiene and Sanitation Transformation methodology was utilized as the key tool in hygiene promotion activities. Three hygiene behaviors were targeted: hand washing with soap, safe feces disposal and boiling drinking water. On average, more than 80% of all children tested were infected with Enterobius vermicularis, Hymenolepis nana, Ascaris lumbricoides, Giardia lamblia or Entamoeba coli. There were statistically significant differences between the three groups for the risk of reinfection rate: the risk of reinfection by parasites was 30% lower in the hygiene promotion group than in the treatment group and 37% lower than in the seasonality group. If properly designed and carried out, hygiene promotion is an effective tool in reducing the risk of intestinal parasite reinfection in children, thus improving children's health in rural communities. PMID- 17418322 TI - Degeneration and possible renewal processes related to the interrenal cells in the head kidney of the stickleback Gasterosteus aculeatus. AB - The ultrastructural aspect of degeneration and recovery processes involving the steroidogenic interrenal cells of the stickleback was studied. Together with the adrenergic cells, the interrenals constitute the adrenal homolog in teleosts. From our study it appears that a process of massive cell death may lead to temporary disappearance of the gland. Moreover, our E.M. observations suggest two main ways, each leading to morphological dedifferentiation of the cells, no longer recognizable as interrenals: the first way involves elimination of organelles and recovery of the nucleus surrounded by a thin rim of cytoplasm; the second involves fragmentation of the cytoplasm by other pyknotic star-shaped interrenals, together with autophagocytosis processes. Our E.M. observations also suggest that the subsequent reconstitution of the tissue can occur in two ways. In the first, the interrenals appear mainly to differentiate from mesenchymatic like electron-light cells, while in the second, the new interrenals appear mainly raising from some macrophagic electron-dense cells. Some data obtained with Mallory's trichrome staining of histological sections, and localization of the enzyme 3beta hydroxysteroid dehydrogenase in thin sections, support the above mentioned results. A hypothesis is advanced on the origin of the electron-dense differentiating interrenals, and a possible role of dedifferentiated cells in restoration of the interrenal gland is also discussed. PMID- 17418325 TI - Arthroscopic repair of massive rotator cuff tears with stage 3 and 4 fatty degeneration. AB - PURPOSE: The purpose of this study was to assess the functional results of arthroscopic repair of massive rotator cuff tears in patients who had stage 3 and 4 fatty degeneration of the rotator cuff musculature, according to the Goutallier scale. METHODS: From January 1997 to December 2001, 22 patients with massive rotator cuff tears and Goutallier stage 3 or 4 fatty degeneration of the infraspinatus, with a mean age of 66.5 +/- 9.26 years, underwent arthroscopic rotator cuff repair and were available for follow-up. The average tear size was 4.8 +/- 0.85 cm in medial-to-lateral width and 6.2 +/- 1.53 cm in anterior-to posterior length, with an approximate tear size area of 30.0 +/- 10.0 sq cm, and involved 2 tendons (supraspinatus and infraspinatus) or 3 tendons (supraspinatus, infraspinatus, and subscapularis) in each case. The mean time from surgery to follow-up was 39.3 months (range, 24-60 months). In addition to comparison of preoperative and postoperative range of motion, strength and University of California, Los Angeles (UCLA) score, outcomes were also assessed with the Constant score. RESULTS: The increase of mean active forward elevation was 53.7 degrees (preoperative: 103.2 degrees and postoperatively: 156.9 degrees ). The gain of mean active external rotation was 19.1 degrees (preoperative: 35.7 degrees and postoperative: 54.8 degrees). The gain of mean external rotation power was 1.9 (preoperative: 2.3 and postoperative: 4.2). The improvement in the UCLA score was 17.2 points (preoperative: 12.3 and postoperative: 29.5). The mean postoperative Constant score was 74.8/100, and the weighted postoperative Constant score was 88.5/100. In 5 patients with fatty degeneration greater than 75% (advanced stage 4), results were less dramatically improved than in 17 patients with fatty degeneration of 50% to 75%, all of whom exhibited clinical improvement. However, clinical improvement was achieved even in 2 of 5 patients with advanced stage 4 involvement. CONCLUSIONS: Arthroscopic rotator cuff repair in patients with grade 3 or 4 fatty degeneration (> or =50%) can provide significant functional improvement. Those with 50% to 75% fatty degeneration showed a much greater degree of improvement (with all 17 cases exhibiting beneficial postoperative increases in their UCLA scores ranging from 12 to 26 points) than those with >75% fatty degeneration (with only 2 of 5 cases showing an increase of 10 or more points in their UCLA scores). However, clinical improvement was observed in 86.4% of cases that would have been classified as likely to fail by the Goutallier criteria. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418326 TI - Cyclic load testing of biodegradable suture anchors containing 2 high-strength sutures. AB - PURPOSE: The purpose of this study was to test 4 different biodegradable suture anchors threaded with 2 high-strength sutures under cyclic loading conditions in humeral cadaveric specimens divided into 2 different age groups. METHODS: Thirty two paired human cadaveric humeri were stripped of all soft tissue. Two groups were studied: group 1, in which the mean age was 54 years, and group 2, in which the mean age was 70 years. We placed 1 suture anchor at 3 humeral sites per bone (anterior, middle, and posterior greater tuberosity). We tested 24 specimens using each of 4 anchors: TwinFix AB (Smith & Nephew Endoscopy, Andover, MA), BioZip (Stryker Endoscopy, San Jose, CA), Bio-Corkscrew FT (Arthrex, Naples, FL), and SpiraLok (DePuy Mitek, Raynham, MA). The anchor's sutures were grasped with an Instron clamp (Instron, Canton, MA), preloaded, and cycled from 10 to 60 N 500 times, followed by destructive testing. The mean displacement at 500 cycles, yield loads, failure modes, and ultimate loads were recorded. RESULTS: Most cyclic motion occurred during the first 100 cycles. More motion occurred in older bones than in younger bones (P < .05). The mean yield loads were greater for the young group for the SpiraLok anchors than for Bio-Corkscrew FT anchors in the young and old groups (P < .001), TwinFix anchors in the old group (P < .05), and BioZip anchors in the old group (P < .05). The ultimate failure loads for SpiraLok anchors in the young group were greater than for Bio-Corkscrew FT anchors in the young and old groups and BioZip anchors in the old group (P < .05). In group 1 TwinFix AB (P = .01) and BioZip (P = .02) ultimate loads were statistically greater than that for Bio-Corkscrew FT. The TwinFix AB failed by anchor pullout. The Bio-Corkscrew FT failed by eyelet pullout. The BioZip and SpiraLok pulled out in older bone and experienced eyelet breakage in younger bone. None of the 4 anchors reached 5 mm of displacement even after 500 loading cycles. CONCLUSIONS: Most of the displacement occurred in the first 100 cycles. Of all anchors tested, the Bio-Corkscrew FT recorded the lowest displacement after 500 cycles (P < .05). The SpiraLok had the highest ultimate load of all anchors tested (P < .01). CLINICAL RELEVANCE: Rotator cuff anchors perform differently in younger humeral bone than in older humeral bone. Most displacement with cyclic loading occurring between the anchor and bone takes place in the first 100 cycles. Anchors in older bones can be expected to fail at lower loads. PMID- 17418327 TI - Patient perceptions of open and arthroscopic shoulder surgery. AB - PURPOSE: There has been a significant increase in surgeon as well as patient interest in arthroscopic treatment of shoulder pathology. We hypothesize that patients have a strong preference for arthroscopic shoulder surgery. Currently, there are no data in the literature reporting on how people perceive open versus arthroscopic shoulder surgery. METHODS: Prospectively, a total of 202 patients were administered a questionnaire at the offices of 2 fellowship-trained shoulder surgeons, 1 located at a tertiary referral center (center A) and 1 in private practice (center B). The questionnaire asked a series of questions about specific factors when choosing an orthopaedic surgeon and the preference for arthroscopic or open surgery. In addition, 7 short-term and long-term outcomes were listed, and the patient was asked to choose whether open or arthroscopic surgery would provide the best outcome or whether he or she felt that there was no difference. RESULTS: Among patients at center A, 88% would prefer arthroscopy if they were to have shoulder surgery, as would 96% of patients at center B. Among patients at center A, 14% would avoid shoulder surgery if the only option was open surgery, as would 25% at center B. The reputation of the surgeon and institution were the primary factors in choosing a surgeon. Recommendation of a former patient and years of surgeon experience were less important factors for patient selection of a shoulder surgeon. CONCLUSIONS: The study shows that the vast majority of patients perceive multiple advantages of arthroscopic shoulder surgery despite a lack of published data to support many of these assumptions, particularly with regard to expected functional outcome of arthroscopic versus open shoulder surgery. CLINICAL RELEVANCE: Shoulder surgeons should be aware of these perceptions when discussing treatment options with patients. A continued effort should be made to emphasize known scientific data when considering the relative risks and benefits of arthroscopic and open shoulder surgery. PMID- 17418328 TI - Arthroscopic Bankart repair using Knotless or BioKnotless suture anchors: 2- to 7 year results. AB - PURPOSE: This study was performed to evaluate the results of arthroscopic Bankart repair, by use of Knotless or BioKnotless suture anchors (DePuy Mitek, Raynham, MA), for traumatic anterior shoulder instability. METHODS: We performed a retrospective evaluation of 73 consecutive patients with traumatic anterior instability of the shoulder treated with arthroscopic Bankart repair by use of metallic Knotless or BioKnotless suture anchors. No additional capsular plication, thermal modification, or interval closure was performed in any patient. The senior author performed all procedures. Independent examiners performed physical examinations. Self-assessment questionnaire evaluations were completed, and preoperative and postoperative American Shoulder and Elbow Surgeons and Rowe scores are reported. RESULTS: Results at a minimum of 2 years' follow-up (range, 2 to 7 years) are reported for 72 patients (57 male and 15 female patients) available for follow-up evaluation. One patient was lost to follow-up. Of the patients, 5 (6.9%) had post-repair instability (3 dislocations and 2 subluxations). The mean postoperative loss of external rotation, at 90 degrees of abduction, was 1 degrees . All patients who had postoperative instability were aged 22 years or younger. The post-repair instability rate in this age group was 13.5% (5/37). No failures occurred in patients aged over 22 years. Of the post-repair dislocations, 3 (7.5%) were in patients involved in contact or collision sports. All 5 failures occurred early, within 2 years of the index surgery. Revision arthroscopic repairs via Knotless or BioKnotless suture anchors were performed in 3 of 4 patients, and a Latarjet procedure was also performed in the fourth patient. The fifth patient refused further intervention. All of the revision shoulders remained stable at the latest follow-up, 3 of which had at least 2 years of follow-up. CONCLUSIONS: Arthroscopic Bankart repair via Knotless or BioKnotless suture anchors showed a recurrence rate of 6.9%. Using Knotless or BioKnotless suture anchors provides satisfactory results with a low recurrence rate, minimal loss of motion, and reliable functional return, even in contact and collision athletes. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418329 TI - Arthroscopic repair of the acetabular labrum: a histologic assessment of healing in an ovine model. AB - PURPOSE: Recently, arthroscopic repair has been advocated for the treatment of labral tears. The purpose of this study was to document the histologic healing process in an arthroscopically repaired acetabular-labral detachment in an ovine model. METHODS: A total of 10 skeletally mature female sheep underwent unilateral arthroscopic labral repair. A 1.5-cm-long incision was made at the junction of the labrum and acetabulum. The labral detachment was then repaired arthroscopically with a single suture anchor. Postoperatively, the animals were confined to small indoor pens, and no form of immobilization was used. After 12 weeks, the animals were killed and the labral repair evaluated via routine histology. Normal labra from 3 nonoperative animals were also processed for histology. RESULTS: The acetabular labrum of the sheep is an avascular, fibrocartilaginous structure similar in anatomic design to that of human beings. All arthroscopically repaired labra appeared stable and grossly healed at 12 weeks. Histologically, labral healing appeared to occur via fibrovascular scar tissue originating from the joint capsule or the exposed bony attachment of the labrum (or both). However, the labral healing was incomplete in all specimens, with a shallow, superficial cleft remaining at the junction of the labrum and the articular surface of the acetabulum. CONCLUSIONS: Arthroscopically repaired acetabular-labral lesions in sheep are capable of healing via fibrovascular repair tissue or direct reattachment via new bone formation (or both). CLINICAL RELEVANCE: The ability of labral detachments to heal in this animal model provides the rationale for additional investigations into the clinical efficacy of arthroscopic labral repair in human beings. PMID- 17418330 TI - Autologous chondrocyte implantation in chondral defects of the knee with a type I/III collagen membrane: a prospective study with a 3-year follow-up. AB - PURPOSE: The clinical results after autologous chondrocyte implantation (ACI) with a collagen membrane using sequential objective patient evaluation (clinical examination and magnetic resonance imaging [MRI]) are inadequately reported. This prospective study was performed to determine the results after ACI at different time intervals over a period of 36 months. METHODS: Between 2000 and 2002, 63 patients (mean age, 34 years) with full-thickness chondral lesions of the knee underwent an autologous chondrocyte implantation and were evaluated preoperatively and at 6, 18, and 36 months after surgery. The chondrocyte suspension within the defect was covered with a type I/III collagen membrane. Depending on the localization of the defects (femoral condyles, trochlea, and retropatellar), the patients were assigned to three different groups. Exclusion criteria were meniscal pathologies, axial malpositioning, and ligament instabilities. Baseline clinical scores (modified Cincinnati knee score and the International Cartilage Repair Society score) were determined and compared with follow-up data by using the paired Wilcoxon test. RESULTS: The ICRS and modified Cincinnati score showed significant improvement (P < .01) in all time intervals between preoperative and 6, 18, and 36 months after surgery. There was no significant difference in the final outcome between different defect localizations (P > .2). The Pearson coefficient of correlation between clinical and MRI scores was 0.73 and significant at the 0.01 level. There was no patient with a symptomatic graft hypertrophy. CONCLUSIONS: ACI is an effective method in the treatment of isolated cartilage defects in the knee. Graft hypertrophy can be avoided by using a collagen membrane. Significant improvement (P < .001) occurs still between 18 and 36 months after surgery. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418331 TI - Meniscus allograft transplantation: ten-year results of cryopreserved allografts. AB - PURPOSE: We report the results of cryopreserved meniscus allograft transplantations with 10 or more years of follow-up. METHODS: Fourteen medial and 8 lateral meniscus allografts were evaluated with a mean follow-up of 141 months (range, 115 to 167 months). The clinical outcome and failure rate was evaluated by use of a Lysholm score and modified pain score in 22 patients. The results of radiographic and magnetic resonance imaging (MRI) analysis were reported in 15 and 7 patients, respectively. RESULTS: Overall, 25% of medial allografts and 50% of lateral allografts failed. The combined failure rate was 35%. There was a 90% improvement in Lysholm scores, as well as pain scores. There were no discernible Lysholm or pain score differences for both lateral and medial allografts. Furthermore, there was no discernible difference in both Lysholm and pain scores between bone plug and soft-tissue methods of graft fixation. Ten of fifteen allografts showed radiographic joint space narrowing, and twelve had progression of degenerative joint disease. On MRI, all grafts had moderate meniscus shrinkage and five had grade III signal intensities. Eighty-five percent of patients underwent subsequent procedures, 5 of whom required total allograft resection and 2 of whom required partial allograft resection. One allograft required repair. CONCLUSIONS: Although transplantation of cryopreserved allografts improved knee pain and function, the average knee function was fair at long-term follow-up. Fifty-five percent of allografts failed when failure criteria for second-look surgery, knee improvement surveys, and MRI were added to Lysholm and pain score failures. The protective benefits of meniscus allografts remain debatable, and inferences cannot be made from this study. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418332 TI - Arthroscopic all-inside repair using the Meniscus Arrow: long-term clinical follow-up of 113 patients. AB - PURPOSE: A clinical trial was carried out to investigate the long-term outcome of Meniscus Arrow (Bionx Implants, Blue Bell, PA) repair. METHODS: In a case series 113 consecutive patients with an arthroscopic all-inside Meniscus Arrow repair were evaluated. The mean age was 30 years, and the mean length of follow-up was 6 years. Of the patients, 84% were available for follow-up. Repairs were performed in either the medial (81%) or lateral (19%) posterior horn in only the red-red or red-white meniscal zone. Concomitant anterior cruciate ligament reconstruction was performed in 66% of patients. RESULTS: Of the patients, 28% showed a retear of the meniscus and had to undergo arthroscopic revision with partial meniscectomy. Simultaneous anterior cruciate ligament reconstruction had no significant influence on the revision rate. In patients who did not undergo revision, the mean Lysholm score was 91 points and the mean Cincinnati knee score was 93 points. The International Knee Documentation Committee classification was A or B in 90% of patients and C in 10%. At revision surgery, 2 patients showed distinct femoral cartilage damage caused by the head of the Meniscus Arrow. CONCLUSIONS: The Meniscus Arrow shows a high clinical failure rate of 28.4%, especially given that the use of the device was restricted to tears in the red red and red-white zone of the meniscus, which would be expected to have a good healing potential. More than 80% of all failures occurred during the first 3 postoperative years, suggesting that the initial refixation potential of the Meniscus Arrow is low. Device-specific complications required additional operative treatment. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418333 TI - Mechanical properties of patellar tendon allografts subjected to chemical sterilization. AB - PURPOSE: We tested the hypothesis that the preimplantation mechanical properties of BioCleanse-treated bone-patellar tendon-bone (BPTB) allografts are not significantly different from those of untreated specimens. METHODS: For this controlled laboratory study, specimens were harvested as central third or hemi BPTB units from both knees of 17 cadaveric tissue donors (11 men and 6 women) aged 19 to 88 years. Donor-matched specimens (20 per group) were randomly assigned to either BioCleanse-treated or untreated control groups. Specimens were subjected to 25 minutes of pretensioning at 89 N and then cyclically loaded under longitudinal tension between 50 N and 250 N for 1,000 cycles at 1 Hz, followed by ramp to failure at 50 mm/min. RESULTS: No statistically significant difference was found between untreated and BioCleanse-treated specimens in stiffness (235.3 +/- 37.6 N/mm v 222.3 +/- 53.4 N/mm, P = .37), cyclic creep (0.38 +/- 0.42 mm v 0.40 +/- 0.26 mm, P = .81), maximum force (1,685.7 +/- 471.6 N v 1,807.0 +/- 657.8 N, P = .47), or ultimate stress (29.0 +/- 9.8 MPa v 29.0 +/- 12.8 MPa, P = .98). CONCLUSIONS: The preimplantation mechanical properties of BPTB allografts treated with BioCleanse are not significantly different from those of untreated controls. CLINICAL RELEVANCE: This laboratory study compares the biomechanical properties of chemically treated allografts, which are currently being used in anterior cruciate ligament reconstruction, with those of nontreated fresh-frozen allografts. PMID- 17418334 TI - Arthroscopic treatment of nonunited anterior cruciate ligament tibial avulsion fracture with figure-of-8 suture fixation technique. AB - PURPOSE: The purpose of this study was to evaluate the clinical results of arthroscopic treatment of nonunited anterior cruciate ligament (ACL) tibial avulsion fracture with a figure-of-8 suture fixation technique. METHODS: Nineteen cases of nonunited ACL tibial avulsion fracture were treated with an arthroscopic figure-of-8 suture fixation technique. The patients were followed up and evaluated according to the International Knee Documentation Committee, Lysholm, and Tegner rating scales. RESULTS: The time interval between the previous injury and this operation was 1 to 8 years, with a mean of 3.6 years. Eighteen patients were followed up for more than 2 years (range, 24 to 30 months; mean, 26 months). The fracture was united within 3 months after surgery in all cases. At the latest follow-up, there was neither extension nor flexion limitation in all patients. With the exception of 1 patient who had a 1 degrees positive Lachman test with a firm end-point, all of the patients had a negative Lachman test. The side-to-side difference in knee anterior laxity was 0 to 2 mm in 17 patients and 4 mm in 1 patient. The Lysholm score was 91 to 100, with a mean of 96.7 +/- 1.9 (compared with 74.3 +/- 4.6 before surgery, P < .05). Fifteen patients returned to their former activity level. The Tegner scores before injury, preoperatively, and at latest follow-up were 6.1 +/- 0.6, 3.6 +/- 1.0, and 6.0 +/- 0.8, respectively; there was no statistically significant difference between the preinjury and postoperative Tegner scores (P > .05). According to the International Knee Documentation Committee scale, 17 patients' knees were graded as normal and 1 patient's knee was nearly normal. CONCLUSIONS: Arthroscopic treatment of nonunited ACL tibial avulsion fracture with a figure-of-8 suture fixation technique can ensure fracture healing and restore the stability and function of the joint in most patients. LEVEL OF EVIDENCE: Level IV, therapeutic case series. PMID- 17418335 TI - Anterior cruciate ligament reconstruction with a porcine xenograft: a serologic, histologic, and biomechanical study in primates. AB - PURPOSE: This study proposes treatment methods to provide a mechanically competent, immunocompatible, and sterile porcine graft for human knee ligament reconstruction. METHODS: The anterior cruciate ligament (ACL) was reconstructed by using treated porcine patellar tendon grafts or controls of untreated porcine grafts or allografts in 20 rhesus monkeys. Animals were stratified into 2-, 6-, and 12-month postreconstruction cohorts. Serologic and histologic assessments were performed to evaluate host immunological and cellular response. Healing and functional integrity of the ACL reconstructions were assessed by tensile biomechanical testing. RESULTS: Untreated porcine grafts were acutely resorbed and rejected, whereas treated porcine grafts and allografts were incorporated by the host as functional grafts. Temporal histologic assessment of treated porcine grafts and rhesus grafts revealed gradual host cellular infiltration and graft collagen remodeling through a similar mechanism of ligamentization. Biomechanical evaluations support graft functional integration with no difference between allograft and treated graft reconstructions. CONCLUSION: Rhesus allograft and treated porcine grafts presented with similar healing profiles in a long-term evaluation of ACL reconstruction. CLINICAL RELEVANCE: Immunochemical modification and sterilization of porcine patellar tendon grafts may improve initial biocompatibility and long-term functionality of xenografts in musculoskeletal applications. PMID- 17418336 TI - Arthroscopic reconstruction of the posterior cruciate ligament by using a quadriceps tendon autograft: a minimum 5-year follow-up. AB - PURPOSE: We prospectively assessed 22 consecutively treated patients to determine the effectiveness and safety of arthroscopically assisted posterior cruciate ligament (PCL) reconstruction by using a quadriceps tendon autograft. METHODS: Twenty-two patients with isolated PCL injury who underwent PCL reconstruction with a quadriceps tendon autograft were enrolled in the prospective study. The average follow-up period was 66 months (range, 60-76). Follow-up included Lysholm knee scores, Tegner activity scores, International Knee Documentation Committee (IKDC) score, thigh muscle assessment, and radiographic assessment. RESULTS: The mean preoperative Lysholm score for 22 knees was 67 (range, 50 to 75), and the mean postoperative Lysholm score was 89 (range, 75 to 98). Nineteen of 22 patients (86%) displayed good or excellent results in the final assessment. The mean preoperative Tegner score for 22 knees was 3 (range, 2 to 5), whereas the mean postoperative Tegner score was 6 (range, 3 to 9). There were statistically significant improvements in Lysholm score (P = .009), Tegner score (P = .039), postoperative KT-1000 arthrometer (MEDmetric, San Diego, CA) scores (P = .006), final IKDC rating (P = .035), and thigh atrophy and muscle strength (P < .05) when compared with preoperative data. Regarding IKDC final rating, 82% of the patients (18 of 22) were assessed as normal or nearly normal (grade A or B). CONCLUSIONS: After follow-up for more than 60 months, the analytical results showed patients achieved satisfactory function after PCL reconstruction by using a quadriceps tendon-patellar bone autograft. This study suggests that a quadriceps tendon autograft is sufficiently large and strong and can achieve good ligament function after reconstruction. LEVEL OF EVIDENCE: Level IV, therapeutic study. PMID- 17418337 TI - Postarthroscopic osteonecrosis of the knee. AB - Little is known about the etiology of postarthroscopic osteonecrosis of the knee. Its prevalence is probably very low. The most important differential diagnosis is pre-existing and undiagnosed early-stage spontaneous osteonecrosis of the knee. From the medicolegal point of view, orthopaedic surgeons need to be aware of the diagnostic pitfalls in differentiating between postarthroscopic osteonecrosis of the knee and spontaneous osteonecrosis of the knee, and they must understand that both may be unpreventable conditions. The purpose of this report is to review the presumable pathophysiology and the clinical and radiographic features as well as the pitfalls in diagnosing postarthroscopic osteonecrosis of the knee. PMID- 17418338 TI - The Di Giacomo technique: simplified suture passing in SLAP repair. AB - A 30 degrees arthroscope is introduced via the posterior soft spot portal, and an anterosuperior portal is created with the use of a 7-mm disposable cannula. The anterosuperior portal is used for instrumentation. An 18-gauge spinal needle is passed via the portal of Neviaser and the rotator cuff into arthroscopic view above the superior labrum. A No. 1 polydioxanone suture (PDS; Ethicon, Somerville, NJ) is advanced through the spinal needle. An arthroscopic retriever or meniscal clamp is used to retrieve the free end of the suture and bring it out through a small anterior stab wound. A suture anchor is inserted via the anterosuperior portal into the superior neck of the glenoid. The more medial limb of the No. 2 Ethibond suture (Ethicon) from the suture anchor is retrieved with the inferior limb of the No. 1 PDS suture, and both are brought out through the anterosuperior cannula. The opposite end of the No. 1 PDS suture is then manually pulled, while, under direct arthroscopic visualization, the No. 2 Ethibond suture, now tied to the opposite end of the PDS, is pulled through the superior labral tissue. That anchor suture is retrieved and is placed outside the cannula that contains the other anchor suture. Standard arthroscopic knot tying is then employed. PMID- 17418339 TI - Arthroscopic rotator cuff repair using a triple-loaded suture anchor and a modified Mason-Allen technique (Alex stitch). AB - Surgical repair of the rotator cuff must have good resistance and should restore the tendon footprint. To attain this goal, a stitch with a strong biomechanical profile that avoids tissue strangulation should be used. We describe an arthroscopic suture technique undertaken to repair rotator cuff tears with a single triple-loaded suture anchor. The technique consists of a combination of a horizontal mattress and 2 vertical simple sutures that are positioned medial to the mattress suture. The suture anchor used is the 5-mm self-tapping ThRevo (Linvatec). This anchor is loaded with 3 sutures: 2 No. 2 nonabsorbable braided polyester sutures of different colors and a central high-strength No. 2 polyethylene suture. The shape of the anchor eyelet permits all 3 sutures to glide freely. A modified Mason-Allen technique (Alex stitch) that combines a horizontal side-to-side suture and 2 simples sutures as vertical loops is used. With use of the Spectrum suture passing device and shuttle relay system (Linvatec), both limbs of the centrally located polyethylene suture are passed through the cuff from bottom to top, approximately 1 cm from the tendon edge. This suture is not immediately tied. Next, with use of the same system, the other 2 sutures are placed medially and over the previous horizontal suture. Simple sutures are placed at an approximately 30 degrees angle from the center of the anchor; 1 is placed anterior and the other posterior. The sutures are tied through the lateral portal. The mattress horizontal central stitch is always tied first, followed by the 2 vertical sutures. The horizontal mattress suture serves as a "rip stop stitch" and theoretically reduces the possibility of cutting out of the simple sutures. PMID- 17418340 TI - Recurrent and chronic complete ruptures of the proximal origin of the hamstring muscles repaired with fascia lata autograft augmentation. AB - Hamstring injuries are common, especially among athletes. A complete rupture of the proximal hamstring muscles requires surgical intervention. In this report we describe a reconstruction method for a complete proximal hamstring rupture using fascia lata autograft augmentation in addition to suture anchors. This method can be advocated in cases in which the primary repair has failed or in chronic injuries where a large defect between the distally retracted tendons and the ischial tuberosity prevents anatomic reinsertion. In our technique, a muscle tendon flap is first created from the retracted tendon stump, turned proximally, and fixed to the ischial tuberosity by suture anchors. The fascia lata graft is then fixed from the midpart to the ischial tuberosity via the same sutures. The other sleeve of the graft is folded on the ventral side of the ruptured tendon stump and fixed by use of absorbable sutures. Then the other sleeve is folded on the dorsal side and fixed in the same manner. Finally, the fixation can still be reinforced with additional absorbable sutures passing through both sleeves of the graft, as well as the muscle-tendon bridge and the tendon stump. PMID- 17418341 TI - Anterior cruciate ligament reconstruction using compressed bone-hamstring-bone graft. AB - Strong initial fixation of the graft is essential for successful early rehabilitation before full graft incorporation. The modified bone-hamstring-bone (BHB) composite graft procedure is a hybrid method that lacks the disadvantages of the bone-patellar tendon-bone and semitendinosus and gracilis tendon methods while retaining the advantages of these methods. We describe the modified BHB composite graft procedure used by our surgical team. Two cancellous bone cylinders, 2 cm long and 8 mm in diameter, were taken out of the tibial tunnel and fixed to both ends of the graft by use of sutures. One end of the graft was put into a pressing device to achieve a graft diameter of 9 or 10 mm, and then the other end was pressed. By doing so, very strong end parts of the graft were obtained. The graft was then passed through the tibia into the femoral socket, and once the graft had been properly positioned in the tunnel, it was fixed with round cannulated interference screws (RCI; Smith & Nephew Endoscopy, Andover, MA). This technique of BHB graft preparation allows precise and strong early fixation, promotes biologic fixation, and does not cause patellofemoral joint disorders. PMID- 17418342 TI - Fluoroscopic-based navigation-assisted placement of the tibial tunnel in revision anterior cruciate ligament reconstruction. AB - With the increasing number of primary anterior cruciate ligament (ACL) reconstructions, revisions are more frequent. The most common cause of technical failure in primary ACL reconstruction is the bone tunnel misplacement. An incorrectly positioned bone tunnel or tunnel enlargement interferes with correct tunnel placement, which is critical for successful restoration of the knee kinematics. We describe a technique using fluoroscopic-based navigation systems to place the tibial tunnel in an ideal position with accuracy and reproducibility in the revision ACL reconstruction. With a reference frame attached to the middle part of the tibia, the conventional tibial guide with a tracking device provides surgeons with real-time visual guidance in multiple image planes. In 3 revision cases with incorrect tibial tunnels and without bone enlargement, the new tibial tunnel could be created in the ideal position under the control of the navigation system. Computer navigation and virtual ligament reconstruction enable surgeons to control bone tunnel positioning during technically demanding revision ACL cases. PMID- 17418343 TI - Arthroscopic posterior cruciate ligament augmentation using an autogenous hamstring tendon graft and the posterior-posterior triangulation technique. AB - We describe a modification of the currently practiced arthroscopic posterior cruciate ligament (PCL) reconstruction techniques. We augmented the injured PCL with quadrupled autogenous hamstring tendon grafts using the arthroscopic posterior-posterior triangulation method and a single-incision transtibial approach. The tibial drill guide was introduced through the anteromedial portal and positioned onto the fossa for the PCL on the tibia under arthroscopic guidance from the posterolateral portal. The femoral tunnel was made 10 mm posterior to the articular cartilage of the medial femoral condyle by use of the anterior cruciate ligament Beath pin (Arthrex, Naples, FL) introduced through the far-inferior anterolateral portal. The tendon graft was positioned in the knee joint by use of the tibial and femoral double-folded silk loops that traversed the bony tunnels. The graft was fixed by use of bioabsorbable Intrafix screw systems (DePuy Mitek, Raynham, MA) at both the ends. The arthroscopic posterior posterior triangulation method provides adequate exposure of the posterior knee compartment; this allows for convenient instrumentation and safe and accurate placement of the bony tunnels with preservation of the PCL remnants. We believe that retention of the remnant PCL fibers is biologic and contributes to earlier healing and strengthening of the tendon graft. PMID- 17418344 TI - Arthroscopic subtalar arthrodesis: the posterior approach in the prone position. AB - Arthroscopic subtalar arthrodesis, as reported by Tasto, is done in the lateral decubitus position, and the portal sites are lateral. This report describes a new alternative method in which the patient is in the prone position and a posterior 2-portal approach is used, as described by van Dijk et al. The initial debridement and synovectomy are performed with 4- and 5-mm resectors. Debridement and decortication are done posterior to the interosseous ligament because only the posterior facet is fused. Denudation of the articular surfaces is performed with curettes, as well as 4.5- and 5.5-mm burs, to remove 2 mm of subchondral bone. Stabilization in 5 degrees of hindfoot valgus is accomplished with 2 percutaneous cannulated headless screws from the non-weight-bearing portion of the calcaneal tuberosity directed to a point 5 to 10 mm posterior to the anterior margin of the posterior facet. The advantages of this alternative treatment are better intra-articular visualization, more thorough preparation of the fusion site, and minimal bone removal of the lateral side with better control of the arthrodesis position and with less chance of malunion, as well as the possibility to perform a concomitant surgical fusion or debridement of the ankle joint during the same operative procedure with no need for additional portals or orientation. PMID- 17418345 TI - Arthroscopic treatment of a massive acromioclavicular joint cyst. AB - A healthy 75-year-old man presented with a 9-month history of an enlarging painless shoulder mass. On examination, there was a 10 x 10 cm firm mass overlying the acromioclavicular (AC) joint. The active range of motion of the shoulder joint was 130 degrees of forward elevation, 90 degrees of abduction, 60 degrees of abduction and internal rotation to the level of the waist. There was weakness of the rotator cuff. Plain radiographs showed degenerative changes of the AC joint with superior migration of the humeral head. Magnetic resonance imaging showed a large cyst arising from a degenerative AC joint with an associated major tear of both the supraspinatus and infraspinatus. Arthroscopy of the AC joint and debridement of the cyst was performed in the lateral decubitus position. The major rotator cuff tear of supraspinatus and infraspinatus was confirmed. A conservative subacromial decompression and AC joint excision arthroplasty was performed. As the superior AC ligament was opened, dark brown gelatinous material was seen emanating from the cyst. A thorough debridement of the cyst was performed until the lavage fluid was clear. At last follow-up 6 months after surgery, the shoulder remained asymptomatic with no evidence of recurrence of the cyst. Arthroscopic treatment of a massive AC joint cyst allows assessment of associated pathology, is minimally invasive, and allows early rehabilitation. PMID- 17418346 TI - Non-Hodgkin's lymphoma of the knee diagnosed by arthroscopy. AB - We report a case of non-Hodgkin's lymphoma of the knee diagnosed by arthroscopy in an otherwise healthy, active 31-year-old woman. Plain radiographs were unremarkable and magnetic resonance imaging findings indicated the presence of a posterior horn medial meniscal tear. The patient underwent arthroscopy for a planned partial medial meniscectomy. However, on joint visualization, hyperemic hypertrophic synovitis was found. Tissue samples were collected and sent for histopathologic testing that revealed diffuse large B cells. The patient was referred to the oncology department where 2 lesions on her lung were also found. Malignant disease should be in the mindset of the orthopaedic surgeon when a patient presents with constant pain, an unusual history, atypical response to conventional treatment, or a diagnostic dilemma. PMID- 17418347 TI - Localized pigmented villonodular synovitis: arthroscopic treatment of a lesion arising from the quadriceps tendon sheath. AB - Pigmented villonodular synovitis (PVNS) is a rare benign proliferative growth of synovium of obscure etiology with a wide spectrum of clinical presentation. Localized PVNS, also known as giant cell tumor of the tendon sheath, is even more uncommon. Localized PVNS of the knee is a rare, idiopathic condition presenting with symptoms that can mimic other intra-articular pathologies. The condition is usually monoarticular, the knee being the most commonly affected joint. We report a rare case of PVNS arising from the quadriceps tendon sheath. It was successfully treated with arthroscopic intralesional excision. At 18 months' follow-up, there has been no recurrence. PMID- 17418348 TI - The bare spot of the glenoid. PMID- 17418350 TI - An alternative method for "all-inside" anterior cruciate ligament reconstruction. PMID- 17418351 TI - Isolation and characterization of a novel toxin from the venom of the spider Grammostola rosea that blocks sodium channels. AB - This communication reports the chemical and physiological characterization of a novel peptide (GrTx1) isolated from the venom of the "rosean tarantula"Grammostola rosea. This component was one among more than 15 distinct components separated from the soluble venom by high-performance liquid chromatography (HPLC). GrTx1 has 29 amino-acid residues, compactly folded by three disulfide bridges with a molecular weight of 3697 Da. Here we show that this peptide blocks Na(+) currents of neuroblastoma F-11 cells with an IC(50) of 2.8+/-0.1 microM, up to a maximum of about 85% at 10 microM. Moreover, the right shift (+20.1+/-0.4 mV) of the fractional voltage-dependent conductance could be also compatible with a putative "gating-modifier" mechanism. No effects were seen on common K(+) channels, such as K(v)1.1 and 1.4, using concentrations of toxin up to 10 microM. Sequence analysis reveals that GrTx1 is closely related to other spider toxins reported to affect various distinct ion channel functions. A critical analysis of this study suggests the necessity to search for other potential receptor sites in order to establish the preferred specificity of these kind of peptides. PMID- 17418352 TI - [Antagonist muscle coactivation and muscle inhibition: effects on external torque regulation and resistance training-induced adaptations]. AB - This review aims at analysing the influence of antagonist muscle coactivation and muscle inhibition on the ability of the neuromuscular system to produce an external torque and to account for changes in these two mechanisms with resistance training. Indeed, antagonist muscle coactivation and muscle inhibition occur during muscle contraction in order to preserve joint integrity. The origin of these two mechanisms would be both spinal and supraspinal and would tend to decrease with resistance training, which allows, under certain conditions, increasing the external torque developed. However, antagonist muscle coactivation and muscle inhibition depend on the characteristics of movement. Moreover, the origin and the contribution of supraspinal mechanisms to the antagonist muscle coactivation and muscle inhibition processes have to be specified. PMID- 17418353 TI - Modulation of soleus H-reflex by presynaptic spinal mechanisms during varying surface and ankle brace conditions. AB - AIMS: Reflex excitability is modulated in part by presynaptic spinal mechanisms. Presynaptic inhibition may prevent an over-response of the motoneuron pool to afferent information. A paired-reflex depression (PRD) conditioning protocol can be used to monitor reflex plasticity. Manipulation of stance, surface, and external bracing are common methods of rehabilitating and treating lower extremity musculoskeletal injuries. The intent of this study was to evaluate changes in PRD of the soleus H-reflex during single-leg stance under varying stability conditions. METHODS: Seven trials were completed for each condition in ten healthy volunteers (age=23+/-1.8 yr, weight 65.0+/-11.3 kg, height=168.7+/ 28.0 cm). The conditioning stimuli were composed of soleus H-reflex pairs evoked 80 ms apart at an equal intensity. The mean percent decrease of the second H reflex relative to the first represented PRD. RESULTS: A 2 x 2 repeated measures ANOVA (P<0.05) was used to evaluate influence of surface (foam, no foam) and support (semi-rigid ankle brace, no ankle brace) on PRD. Main effects testing revealed a significantly greater soleus PRD (P=.034) for the foam surface (62.5%) compared the flat surface (57.5%). Ankle brace application did not influence soleus PRD (P=0.63). CONCLUSION: The increase in soleus PRD during the foam condition suggests depression of the motoneuron pool. This may lessen postural over-corrections while maintaining upright stance during less stable conditions. No change in PRD during the ankle brace condition suggests that mechanical reinforcement provided an increase in ankle stability, decreasing the demand on the motoneuron pool. PMID- 17418354 TI - Experience with external pump trial prior to implantation for intrathecal baclofen in ambulatory patients with spastic cerebral palsy. AB - OBJECTIVES: To evaluate effectiveness and safety of intrathecal baclofen administration (ITB) testing with continuous infusion via an external pump before the implantation of an internal one in ambulatory spastic patients with cerebral palsy (CP). PATIENTS AND METHODS: Seven CP patients (3 diplegic, 4 quadriplegic - 18.4+/-7.0 years) with a progressive decrease in walking ability were included. Assessments included: Ashworth's scale, Observational Gait Scale (OGS), and GMFM 66. RESULTS: During the ITB test (45-150 microg/24h), spasticity decreased by more than two points on Ashworth's scale (p<0.001) and walking ability improved (median OGS increased from 7 to 9, p1.0 microg/g ww) at all CRB sites except the Gila River at Hayden, Arizona. Mercury concentrations were elevated (>0.1 microg/g ww) in fish from the Yampa River at Lay, Colorado; the Green River at Ouray National Wildlife Refuge (NWR), Utah and San Rafael, Utah; the San Juan River at Hogback Diversion, New Mexico; and the Colorado River at Gold Bar Canyon, Utah, Needles, California, and Imperial Dam, Arizona. Concentrations of p,p'-DDE were relatively high in fish from the Gila River at Arlington, Arizona (>1.0 microg/g ww) and Phoenix, Arizona (>0.5 microg/g ww). Concentrations of other formerly used pesticides including toxaphene, total chlordanes, and dieldrin were also greatest at these two sites but did not exceed toxicity thresholds. Currently used pesticides such as Dacthal, endosulfan, gamma-HCH, and methoxychlor were also greatest in fish from the Gila River downstream of Phoenix. Total polychlorinated biphenyls (PCBs; >0.11 microg/g ww) and TCDD-EQs (>5 pg/g ww) exceeded wildlife guidelines in fish from the Gila River at Phoenix. Hepatic ethoxyresorufin O-deethylase (EROD) activity was also relatively high in carp from the Gila River at Phoenix and in bass from the Green River at Ouray NWR. Fish from some sites showed evidence of contaminant exposure as indicated by fish health indicators and reproductive biomarker results. Multiple health indicators including altered body and organ weights and high health assessment index scores may be associated with elevated Se concentrations in fish from the Colorado River at Loma, Colorado and Needles. Although grossly visible external or internal lesions were found on most fish from some sites, histopathological analysis determined many of these to be inflammatory responses associated with parasites. Edema, exophthalmos, and cataracts were noted in fish from sites with elevated Se concentrations. Intersex fish were found at seven of 14 sites and included smallmouth bass (M. dolomieu), largemouth bass (M. salmoides), catfish, and carp and may indicate exposure to endocrine disrupting compounds. A high proportion of smallmouth bass from the Yampa River at Lay (70%) was intersex but the cause of this condition is unknown. Male carp, bass, and catfish with low concentrations of vitellogenin were common in the CRB. Comparatively high vitellogenin concentrations (>0.2 mg/mL) were measured in male bass from the Green River at Ouray NWR and the Colorado River at Imperial Dam and indicate exposure to estrogenic or anti-androgenic chemicals. Anomalous reproductive biomarkers including low GSI and gonadal abnormalities (calcifications, edema, and parasites) observed in fish downstream of Phoenix are likely related to the poor water-quality of the Gila River in this area. PMID- 17418377 TI - Aerosol advection and sea salt events in Genoa, Italy, during the second half of 2005. AB - Atmospheric aerosols in the PM(10) fraction have been simultaneously sampled at three sites in the Genoa urban and suburban area during the second half of 2005, and information on the elemental composition has been gathered through energy dispersive X-ray fluorescence. Thanks to the simultaneous measurements and wind information, a few aerosol transport and transformation processes originated from the nearby sea and in the neighbouring Po Valley have been described. Sea salt concentrations at the three sites were well correlated and often related to Southern sector winds; moreover, by examining the Cl/Na ratio at two sites the time scale for Cl depletion in particulate matter has been estimated as 1-1.5 h for the Genoa atmosphere. During a Northerly gale, excess elemental Si concentrations (peaking more than 4 mug m(-3)) were found at two sites, and were ascribed to an unknown local source. Finally, during an 11-day long 'heat wave' large concentrations for total PM(10), dust and secondary compounds have been found; these large concentrations lead to a number of exceedances of air quality standards, and have been ascribed to advection from the Po Valley. PMID- 17418378 TI - Aspiration before immunotherapy injection is required. PMID- 17418379 TI - Expression and regulation of intercellular adhesion molecule-1 on airway parasympathetic nerves. AB - BACKGROUND: Eosinophils cluster along airway nerves in patients with asthma and release eosinophil major basic protein, an antagonist of inhibitory M2 muscarinic receptors on nerves. Blocking M2 function increases bronchoconstriction, leading to airway hyperreactivity. Intercellular adhesion molecule-1 (ICAM-1) mediates eosinophil adhesion to nerves. OBJECTIVE: We investigated mechanisms of ICAM-1 expression by parasympathetic nerves. METHODS: ICAM-1 expression was examined by immunocytochemistry of lung sections from ovalbumin-sensitized and challenged guinea pigs. ICAM-1 was measured in parasympathetic nerves isolated from subjects and guinea pigs and in human neuroblastoma cells by real-time RT-PCR, immunocytochemistry, and Western blot. RESULTS: ICAM-1 was not detected in control airway parasympatheric nerves in vivo or in cultured cells. ICAM-1 was expressed throughout antigen-challenged guinea pig lung tissue and was selectively decreased by dexamethasone only in nerves. ICAM-1 was induced in human and guinea pig parasympathetic nerves by TNF-alpha and IFN-gamma and was inhibited by dexamethasone and by an inhibitor of nuclear factor-kappaB (NF kappaB). In neuroblastoma cell lines TNF-alpha and IFN-gamma-induced ICAM-1 was blocked by an inhibitor of NF-kappaB but not by inhibitors of mitogen-activated protein kinases. Dexamethasone did not inhibit ICAM-1 expression in neuroblastoma cells. CONCLUSIONS: ICAM-1 induced in nerves by antigen challenge and proinflammatory cytokines is sensitive to dexamethasone. ICAM-1 expression is also sensitive to inhibitors of NF-kappaB. Neuroblastoma cells mimic many, but not all, characteristics of ICAM-1 expression in parasympathetic nerves. CLINICAL IMPLICATIONS: Dexamethasone and NF-kappaB inhibitors could prevent eosinophils from adhering to nerves by blocking ICAM-1 expression on parasympathetic nerves, thus protecting inhibitory M2 muscarinic receptors and making this pathway a potential target for asthma treatment. PMID- 17418380 TI - Mechanisms of tissue inhibitor of metalloproteinase 1 augmentation by IL-13 on TGF-beta 1-stimulated primary human fibroblasts. AB - BACKGROUND: TGF-beta induces expression of tissue inhibitor of metalloproteinase 1 (TIMP-1), a potent inhibitor of matrix metalloproteinases that controls extracellular matrix metabolism and deposition. IL-13 alone does not induce TIMP 1, but in combination with TGF-beta it augments TIMP-1 expression. Although these interactions have implications for remodeling in asthma, little is understood regarding the mechanisms controlling TIMP-1 product. OBJECTIVE: To explore the role of Smads and mitogen-activated protein kinase kinase (MEK)-extracellular signal-regulated kinase (ERK) in the TIMP-1 augmentation by IL-13+TGF-beta1 in primary human airway fibroblasts. METHODS: Real-time PCR, Western blot, ELISA, and transient transfection were used to evaluate the mechanisms of TIMP-1 augmentation. RESULTS: IL-13 enhanced TGF-beta1-induced Smad-2 and Smad-3 phosphorylation, transient transfection with dominant-negative Smad-2 or Smad-3 decreased TIMP-1 mRNA expression in the presence of TGF-beta1 and IL-13+TGF-beta1 through inhibition of Smad-2 or Smad-3 phosphorylation. ERK phosphorylation was increased by IL-13 and IL-13+TGF-beta1. MEK-ERK inhibition decreased TIMP-1 mRNA/protein to a greater degree after IL-13+TGF-beta1 stimulation versus TGF beta1 alone. MEK-ERK inhibition also significantly increased Akt phosphorylation under all conditions and decreased Smad-3 phosphorylation in the presence of IL 13+TGF-beta1. In contrast, phosphoinositide-3 kinase-Akt inhibition increased phosphorylation of ERK and Smads, leading to increased TIMP-1. CONCLUSION: These results indicate that IL-13 augments TGF-beta1-induced TIMP-1 expression through increased Smad phosphorylation. These increases occur as TGF-beta1 downregulates IL-13-induced phosphoinositide-3 kinase activation while leaving the positive effect of IL-13-induced ERK on Smad signaling. CLINICAL IMPLICATIONS: This augmentation of TGF-beta1-induced TIMP-1 by IL-13 could contribute to the fibrosis and airway remodeling seen in the presence of T(H)2 inflammation in asthma. PMID- 17418381 TI - The IL-17F signaling pathway is involved in the induction of IFN-gamma-inducible protein 10 in bronchial epithelial cells. AB - BACKGROUND: IL-17F is involved in airway inflammation, but its biologic activity and signaling pathway remain incompletely defined. Interferon-gamma-inducible protein 10 (IP-10) is widely expressed and plays a role in airway inflammatory diseases. OBJECTIVE: We sought to investigate the functional linkage between IL 17F and IP-10 expression in bronchial epithelial cells. METHODS: Bronchial epithelial cells were cultured in the presence or absence of IL-17F, and/or a T(H)1 cytokine, T(H)2 cytokines, proinflammatory cytokines, various kinase inhibitors, or a Raf1 dominant-negative mutant to analyze the expression of IP 10. Moreover, the involvement of p90 ribosomal S6 kinase (p90RSK) and cyclic AMP response element-binding protein (CREB) in IL-17F-induced IP-10 expression were investigated. RESULTS: IL-17F induces the gene and protein expression of IP-10. The addition of IFN-gamma, IL-1beta, and TNF-alpha augmented IL-17F-induced IP-10 expression. The mitogen-activated protein kinase kinase (MEK) inhibitors PD98059, U0126, and Raf1 kinase inhibitor I significantly inhibited its production. In contrast, a p38 inhibitor, a JNK inhibitor, protein kinase C inhibitors, and a phosphatidylinositol 3-kinase inhibitor, showed no inhibitory effect. Furthermore, overexpression of a Raf1 dominant-negative mutant inhibited its expression. Of interest, IL-17F phosphorylated p90RSK and CREB, and transfection of the cells with a short interfering RNA for p90RSK or CREB inhibited its expression, suggesting p90RSK and CREB as novel signaling molecules of IL-17F. CONCLUSION: IL-17F is a potent inducer of IP-10 in bronchial epithelial cells through the activation of the Raf1-MEK1/2-extracellular signal-regulated kinase 1/2-p90RSK-CREB pathway, supporting its regulatory role in airway inflammation. CLINICAL IMPLICATIONS: The IL-17F-IP-10 axis might be a novel and critical therapeutic target for airway inflammatory diseases. PMID- 17418382 TI - Phase I/II trial of intermittent subcutaneous IL-2 administration in pediatric patients with moderate immune suppression: results of Pediatric AIDS Clinical Trials Study 402. PMID- 17418383 TI - Treatment of acute edema attacks in hereditary angioedema with a bradykinin receptor-2 antagonist (Icatibant). AB - BACKGROUND: In hereditary angioedema, bradykinin is assumed to be the most important mediator of edema formation. OBJECTIVE: To assess whether the selective bradykinin receptor-2 antagonist Icatibant is effective in acute edema attacks of hereditary angioedema. METHODS: In this uncontrolled pilot study, 15 patients with 20 attacks were treated with Icatibant. The attacks were analyzed by using a standardized and validated visual analog scale measurement and compared with historical data of untreated attacks. Plasma bradykinin concentration was measured before and 4 hours after intravenous Icatibant treatment. RESULTS: Symptom intensity decreased within 4 hours after administration of Icatibant; the median time to onset of symptom relief was 1.50, 1.42, and 1.13 hours in the intravenous groups and 0.58 and 0.45 hours in the subcutaneous groups, respectively. The median difference in the 10-cm visual analog scale 4 hours after start of treatment was 4.11 cm (95% CI, 1.72-6.07). Compared with untreated attacks, Icatibant treatment reduced the mean (SD) time to onset of symptom relief by 97% from 42 +/- 14 to 1.16 +/- 0.95 hours (all groups combined). Median bradykinin concentration was 7-fold above the norm during acute attacks at 48.5 pmol/L and decreased to 18.0 pmol/L 4 hours after Icatibant infusion or injection. CONCLUSION: Icatibant was effective in treating acute attacks of hereditary angioedema. CLINICAL IMPLICATIONS: This is the first report demonstrating the clinical usefulness of antagonizing bradykinin binding to bradykinin receptor-2 in hereditary angioedema. PMID- 17418384 TI - Fluticasone furoate nasal spray: a single treatment option for the symptoms of seasonal allergic rhinitis. AB - BACKGROUND: Fluticasone furoate (USAN-approved name) is a novel, enhanced affinity glucocorticoid administered in a unique side-actuated device for the management of seasonal allergic rhinitis (SAR). OBJECTIVE: We sought to evaluate the efficacy and safety of once-daily fluticasone furoate nasal spray, 110 microg, in patients aged 12 years or older with fall SAR. METHODS: Patients (n = 299) received fluticasone furoate or placebo for 2 weeks in this double-blind, parallel-group randomized study. Patients evaluated nasal and ocular symptoms using a 4-point categoric scale. Efficacy was assessed on the basis of the mean change from baseline in reflective and instantaneous total nasal symptom scores and reflective total ocular symptom scores. RESULTS: Fluticasone furoate produced significantly greater improvements than placebo in daily reflective total nasal symptom score (-1.473, P < .001; primary end point), morning predose instantaneous total nasal symptom score (-1.375, P < .001), daily reflective total ocular symptom score (-0.600, P = .004), and patient-rated overall response to therapy (P < .001). The onset of therapeutic effect occurred at 8 hours after initial administration. Fluticasone furoate was well tolerated. CONCLUSION: Fluticasone furoate, 110 microg once daily, was effective and well tolerated for the treatment of nasal symptoms of SAR in patients aged 12 years and older. Treatment also produced significant improvements in ocular symptoms. Fluticasone furoate was fast acting, as indicated by an 8-hour onset of action, and provided 24-hour symptom control. CLINICAL IMPLICATIONS: New treatments for the bothersome symptoms of SAR are needed. One such treatment, fluticasone furoate nasal spray, provides effective relief of the symptom profile of SAR. PMID- 17418386 TI - Ovarian function--from the beginning. PMID- 17418387 TI - Undescended ovaries: a clinical review. AB - Undescended ovaries are frequently seen in conjunction with uterine malformations and are typically found during the course of an infertility evaluation. Other important clinical signs may prompt evaluation in an adolescent patient, though this may also be an incidental finding. An understanding of embryologic features is critical for management. It is additionally important to explain to patients and families that this is a normal developmental variant of the ovary that does not require removal in the absence of concerning pathology. PMID- 17418388 TI - Preventing repeat teen pregnancy: postpartum depot medroxyprogesterone acetate, oral contraceptive pills, or the patch? AB - STUDY OBJECTIVE: To prospectively evaluate the repeat teen pregnancy rates, within one year of delivery, among adolescents who choose the contraceptive patch (Ortho Evra) versus oral contraceptive pills (OCP) versus Depot Medroxyprogesterone Acetate (Depo Provera, DMPA) for postpartum contraception. DESIGN: Observational, prospective cohort study. Comparison groups are postpartum teens, who self-select the contraceptive patch (n = 55) versus DMPA (n = 142) versus OCPs (n = 55) immediately postpartum. SETTING: Medical University of South Carolina, a tertiary medical center. PARTICIPANTS: Postpartum teens, 11-19 years old; 72% were African American, and 96% qualified for Medicaid insurance. INTERVENTIONS: A structured telephone interview was performed every 3 months. MAIN OUTCOME MEASURES: The primary outcome measure was a repeat pregnancy within 12 months of the index delivery. Secondary outcome variables were contraceptive continuation rates, reasons for discontinuation, side effects and condom usage. RESULTS: At 1-year follow-up, repeat pregnancy rates were 14.2%, 29.7%, and 31.8% among DMPA, OCP, and patch users respectively (P = 0.02). DMPA users were significantly more likely to be using any form of hormonal contraception 1 year postpartum than patch or OCP users. Condom use was similarly low among all cohorts. CONCLUSION: Adolescents who choose DMPA for postpartum contraception are significantly less likely to become pregnant within 1 year of delivery, as compared to teens who choose OCPs or the patch. PMID- 17418389 TI - Normal ovarian structure and function with normal glucose tolerance in girls with early treatment of classic congenital adrenal hyperplasia. AB - Hyperandrogenism and hyperinsulinism have both been suggested as etiologic factors behind functional ovarian hyperandrogenism or polycystic ovary syndrome. Females with congenital adrenal hyperplasia provide a clinical model to evaluate the contribution of pre- and post-natal hyperandrogenism on ovarian structure and function. STUDY OBJECTIVE: To evaluate glucose tolerance, and structure and androgen production of the ovaries in young females with classic congenital adrenal hyperplasia. DESIGN, SETTING, PARTICIPANTS: Cross-sectional study, including the enrollment of participants, ages 8 to 20 years, recruited from the pediatric endocrinology clinical program of a tertiary pediatric referral center. INTERVENTIONS: Ten participants had oral glucose tolerance testing, adrenal and ovarian androgen measurements, and pelvic ultrasound studies performed. MAIN OUTCOME MEASURES: Presence of altered response to glucose challenge, ovarian hyperandrogenism, or presence of polycystic ovaries by ultrasound. RESULTS: Measurements of fasting blood glucose, post-challenge glucose, and insulin resistance were normal in this sample. There was no evidence of ovarian hyperandrogenism after adrenal suppression with dexamethasone. All participants had normal ovarian structure without evidence of polycystic ovaries. CONCLUSIONS: Females with classic congenital adrenal hyperplasia (21-hydroxylase deficiency) and normal glucose tolerance appear to have normal ovarian structure and function during the peripubertal period. PMID- 17418390 TI - Characteristics of adolescent women who stop using contraception after use at first sexual intercourse. AB - STUDY OBJECTIVE: Identify correlates of contraceptive discontinuation, which if modified, might make teenagers more, not less, effective contraceptors as they age. SETTING: Teen clinic. PARTICIPANTS: Teenagers who used contraception at first intercourse (N = 120). Some "never" used contraception during the 4 months immediately prior to the survey ("contraceptive stoppers"; n = 38). The others (n = 82) did so "always" or "most of the time" ("consistent contraceptive users"). INTERVENTIONS: Questionnaire responses were used to determine univariate and multivariate associations between contraceptive use group and five categories of factors: inability to plan for sex, belief that pregnancy is unlikely to occur, belief that contraceptives are unsafe, inability to negotiate contraceptive use, and lack of desire to remain non-pregnant. MAIN OUTCOME MEASURE: Odds of being a contraceptive stopper. RESULTS: In univariate analyses contraceptive stoppers scored significantly higher on scales that assessed inability to plan for sex, belief that pregnancy is unlikely, and lack of desire to remain non-pregnant. Contraceptive stoppers were also older and more likely to have been sexually active for at least 6 months. In multivariate analyses, those who were sexually active for at least 6 months (odds ratio [OR]: 2.9, confidence interval [95%CI]: 1.1-7.1), those who believed that pregnancy was unlikely (OR: 3.8; 95% CI: 1.7 8.6), and those who lacked the desire to remain non-pregnant (OR: 2.7; 95% CI: 1.4-5.1) were more likely to stop using contraception. CONCLUSIONS: Our findings suggest that teens who use contraception at coitarche stop doing so as they mature sexually because they begin to doubt the necessity and desirability of using contraceptives. Longitudinal studies are needed to determine if such doubts are preventable and if doing so encourages teens to continue to use contraception. PMID- 17418391 TI - Comparison of quality of life scores among non-exercising adolescent females and adolescent dancers with oligomenorrhea and amenorrhea. AB - STUDY OBJECTIVE: To evaluate whether oligomenorrhea and amenorrhea in adolescent females would have a negative effect on the standard quality of life (QOL) scores as compared to eumenorrheic adolescents, and whether such scoring would differ between those with exercise related oligo/amenorrhea or those with hypothalamic/ovarian causes. DESIGN: Prospective cross-sectional observational survey. METHODS: Young dancers were recruited from the Sports Clinic of a collegial School of Dance, and non-dancers were recruited from a hospital-based adolescent gynecology clinic. All subjects completed a structured self-answering questionnaire recording epidemiological data and a 36-item Short Form Health Survey (SF-36). All subjects had a full hormonal profile and pelvic ultrasound to study ovarian morphology. RESULTS: Sixty-six dancers (19 with exercise-related oligo/amenorrhea and 47 eumenorrhoeic) and 90 non-dancers (45 eumenorrhoeic and 45 oligo/amenorrheic) were analyzed, all 16-20 years of age. There was no difference in the scoring between eumenorrheic dancers and non-dancers. Oligo/amenorrheic dancers did not score lower than eumenorrheic dancers. Compared with eumenorrheic subjects or to oligo/amenorrheic dancers, oligo/amenorrheic non dancers had significantly lower QOL scores in the domains of physical functioning (PF) and general health (GH) and vitality (VT). CONCLUSION: Compared with eumenorrheic adolescents, QOL scores were lower in non-exercising adolescents with oligo-amenorrhea, but not those with exercise related oligo/amenorrhea. The negative effects of oligomenorrhea and amenorrhea on quality of life were apparently attenuated if the menstrual dysfunction was related to physical training. PMID- 17418392 TI - Gonadal function in adolescent patients submitted to chemotherapy during childhood or during the pubertal period. AB - STUDY OBJECTIVE: To determine the presence of impaired gonadal function in adolescent patients submitted to chemotherapy during childhood or during the pubertal period. DESIGN: A case series study of 28 patients aged 12 to 19 years with menarche at least 2 years before the study. SETTING: Tertiary care public hospital. PARTICIPANTS: Group I: 14 adolescents previously submitted to chemotherapy during the prepubertal or peripubertal period and with remission of oncologic disease for at least 2 years; Group II: 14 normal adolescents with no previous oncologic disease and with regular menstrual cycles. INTERVENTIONS AND MAIN OUTCOME MEASURES: Pubertal development, menstrual cycles and serum levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were determined during the early follicular phase. RESULTS: There were no differences between the two groups in terms of age at appearance of secondary sexual characteristics or age at menarche. Menstrual irregularity was detected in 7 of the 14 patients in Group I, all 8 of whom presented oligomenorrhea. There were no differences in LH levels between the two groups (P = 0.55), although mean FSH levels were higher in Group I than in Group II (6.71 +/- 2.99 mIU/ml vs. 3.83 +/- 2.01 mIU/ml, P = 0.01). CONCLUSION: Although girls submitted to chemotherapy during the prepubertal or peripubertal period presented normal sexual development, the incidence of oligomenorrhea was higher than expected for their age, and FSH levels, although within normal limits, were higher than those seen in normally cycling girls. PMID- 17418393 TI - Self-induced medical abortion in an adolescent. AB - CASE REPORT: An 18-yr-old female presented to her local pediatric clinic with a 4 day history of vaginal bleeding and cramping. She reported sexual activity with last coitus two weeks prior and intermittent condom usage. Examination revealed blood in the vaginal vault and a 4- to 5-cm uterus, slightly tender to palpation. Urine human chorionic gonadotropin (hCG) was positive. She reported having taken "four little white pills" from a friend, prior to onset of bleeding, "so that she would not be pregnant." Further questioning revealed that the pills were misoprostol. An ultrasound demonstrated no intrauterine or tubal pregnancy. The patient was followed; her bleeding resolved over two weeks and her ss-hCG levels declined steadily. DISCUSSION: There are some studies in the adult literature that address medical self-induction of abortion, and sporadic reports on the Internet and in the lay press. No pediatric literature has been published on this topic in the United States. It is important for clinicians to be aware that such an option exists for pregnant young women when evaluating an adolescent who presents with vaginal bleeding and cramping. PMID- 17418394 TI - Management of ruptured corpus luteum cyst of pregnancy occurring in a 15-year-old girl by laparoscopic surgery with intraoperative autologous blood transfusion. AB - BACKGROUND: Laparoscopic surgery is a minimally invasive procedure with many advantages. However, laparoscopic treatment of ruptured corpus luteum cyst of pregnancy with massive hemoperitoneum occurring in a young girl has not previously been reported. CASE: A 15-yr-old girl presenting with acute abdomen and hemoperitoneum was referred to our department. A urinary pregnancy test was positive and an ultrasound revealed a gestational sac in the uterine cavity, the preoperative differential diagnosis was narrowed to either intrauterine pregnancy with ruptured corpus luteum cyst or heterotopic pregnancy. Emergency laparoscopic surgery was performed to investigate the cause of hemoperitoneum and a diagnosis of ruptured corpus luteum cyst of pregnancy was established. After retrieving pooled blood in the abdominal cavity for intraoperative autologous blood transfusion, the rupture site with active bleeding was laparoscopically sutured and hemostasis was achieved. At the same time, intrauterine pregnancy was electively terminated at the request of the patient and her family. The postoperative course was uneventful. CONCLUSION: Ruptured corpus luteum cyst of pregnancy manifesting massive hemoperitoneum is a rare but life-threatening disorder that can occur even in a young girl. Ovarian conservative treatment can laparoscopically be performed with intraoperative autologous blood transfusion. PMID- 17418395 TI - Polycystic ovary syndrome in adolescents. PMID- 17418396 TI - Management Quandary. Extensive perineal infantile hemangioma with associated congenital anomalies: an example of the PELVIS syndrome. PMID- 17418397 TI - What's your diagnosis? Scaly pubic plaques in a 2-year-old girl--or an "inverse" rash. PMID- 17418398 TI - Effects of sediment organic matter quality on bioaccumulation, degradation, and distribution of pyrene in two macrofaunal species and their surrounding sediment. AB - Sediment dwelling macrofauna (infauna) are important vectors for the transfer of sediment-associated contaminants to higher trophic levels. Sedimenting organic matter constitutes an important food source for all benthic organisms and changes seasonally in terms of quantity and quality. Sediment organic matter (SOM) quality affects organism activity and feeding behaviour, and is therefore also likely to affect contaminant fate in benthic systems. We investigated the impact of SOM quality (enrichment with either labile Tetraselmis sp. or refractory lignin) on the accumulation and metabolism of sediment-associated pyrene in Nereis diversicolor (Annelida) and Amphiura filiformis (Echinodermata), as well as the combined effect of SOM quality and infaunal bioturbation on pyrene distribution and metabolism in the sediment. After 45 d of exposure, SOM quality almost doubled pyrene bioaccumulation in both species, while pyrene metabolism remained unaffected. Metabolites comprised approximately 80% of the total tissue pyrene in N. diversicolor and approximately 40% in A. filiformis. A. filiformis arms contained one fifth of the disk pyrene concentration. Approximately 20% of the pyrene found in A. filiformis arms was found to be covalently bound to, e.g. DNA, RNA or proteins, thus reducing pyrene bioavailability to arm-cropping predators. The sedimentary pyrene distribution and metabolism was species dependent, but correlated poorly with prevailing knowledge on species-specific bioturbation patterns. This was attributed to the comparably high biodegradability of the contaminant thus altering its sorptive characteristics and function as inert tracer. Subduction of pyrene and metabolites occurred, and the fraction of pyrene covalently bound to SOM increased with depth, thereby removing pyrene from the bioavailable pool. Our results imply that bioaccumulation and trophic transfer of sediment-associated PAH should increase following fresh organic matter input, e.g. after sedimentation of phytoplankton blooms. We stress the importance of considering behavioural characteristics of infauna and the trophic situation of the system when assessing fate and effects of sediment-associated contaminants. PMID- 17418399 TI - Lysozyme purification with dye-affinity beads under magnetic field. AB - Magnetic poly(2-hydroxyethyl methacrylate) mPHEMA beads carrying Cibacron Blue F3GA were prepared by suspension polymerization of HEMA in the presence of Fe3O4 nano-powder. Average size of spherical beads was 80-120 microm. The beads had a specific surface area of 56.0m(2)/g. The characteristic functional groups of dye attached mPHEMA beads were analyzed by Fourier transform infrared spectrometer (FTIR) and Raman spectrometer. mPHEMA with a swelling ratio of 68% and carrying 28.5 micromol CibacronBlueF3GA/g were used for the purification of lysozyme. Adsorption studies were performed under different conditions in a magnetically stabilized fluidized bed (i.e., pH, protein concentration, flow-rate, temperature, and ionic strength). Lysozyme adsorption capacity of mPHEMA and mPHEMA/Cibacron Blue F3GA beads were 0.8 mg/g and 342 mg/g, respectively. It was observed that after 20 adsorption-desorption cycle, mPHEMA beads can be used without significant loss in lysozyme adsorption capacity. Purification of lysozyme from egg white was also investigated. Purification of lysozyme was monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 87.4% with recovery about 79.6%. The specific activity of the desorbed lysozyme was high as 41.586 U/mg. PMID- 17418400 TI - Chemical characteristics and immuno-modulating activities of exo-biopolymers produced by Grifola frondosa during submerged fermentation process. AB - The immuno-modulating activities and chemical characteristics of exo-biopolymer (EX-GF) produced by a submerged mycelial culture of Grifola frondosa were studied. The EX-GF was fractionated into EX-GF-Fr.I, II, and III by Sephadex G 100 gel chromatography. Anti-complementary activity of EX-GF-Fr.III was highest (71.1%) among them, and its activation system occurred through both classical and alternative pathways, where the classical pathway found to be major one. Lysosomal enzyme activity and nitric oxide production ability of macrophage were also found to be mediated by EX-GF-Fr.III. The molecular weight of the EX-GF Fr.I, II, and III was estimated to be about 163, 40, and 2.8 kDa, respectively. Total sugar and protein contents of the three fractions were 80.3, 61.9 and 89.3%, and 17.3, 35.2, and 10.7%, respectively. The sugar and amino acid compositions of the EX-GF-Fr.I, II, and III were also analyzed in detail. PMID- 17418401 TI - The effect of 3-hydroxybutyrate on the in vitro differentiation of murine osteoblast MC3T3-E1 and in vivo bone formation in ovariectomized rats. AB - 3-hydroxybutyrate (3HB), one of the degradation products of microbial biopolyesters polyhydroxyalkanoates (PHA), is a high energy metabolic substrate in animals. This study evaluated the effects of 3HB on growth of osteoblasts in vitro and on anti-osteoporosis in vivo. Alkaline phosphatase (ALP) assay, Van Kossa assay and Alizarin S red staining were used to study in vitro differentiation of murine osteoblast MC3T3-E1 cells. The intensity of in vitro cell differentiation measured in ALP was in direct proportion to the concentration of 3HB when it was lower than 0.01 g/L. Calcium deposition, a strong indication of cell differentiation, also showed an obvious increase with increasing 3HB concentration from 0-0.1g/L, evidenced by Alizarin red S staining and Van Kossa assay. RT-PCR also showed significantly higher expression of osteocalcin (OCN) mRNA in MC3T3-E1 cells after 3HB administration. In vivo study using female Wistar rats (3 months old, n=80) allocated into normal, sham operated or ovariectomized (OVX) group that led to decreasing bone mineral density (BMD), bone histomorphometry and biomechanics compared with normal and sham groups, had demonstrated that 3HB increased serum ALP activity and calcium deposition, decreased serum OCN, prevented BMD reduction resulting from OVX. All these led to enhanced femur maximal load and bone deformation resistance, as well as improved trabecular bone volume (TBV%). In conclusion, 3HB monomer containing PHA can be effective bone growth stimulating implant materials. PMID- 17418402 TI - [Lung cancer]. PMID- 17418403 TI - A quantitative model for linking Na+/Ca2+ exchanger to SERCA during refilling of the sarcoplasmic reticulum to sustain [Ca2+] oscillations in vascular smooth muscle. AB - We have developed a quantitative model for the creation of cytoplasmic Ca2+ gradients near the inner surface of the plasma membrane (PM). In particular we simulated the refilling of the sarcoplasmic reticulum (SR) via PM-SR junctions during asynchronous [Ca2+]i oscillations in smooth muscle cells of the rabbit inferior vena cava. We have combined confocal microscopy data on the [Ca2+]i oscillations, force transduction data from cell contraction studies and electron microscopic images to build a basis for computational simulations that model the transport of calcium ions from Na+/Ca2+ exchangers (NCX) on the PM to sarcoplasmic/endoplasmic reticulum Ca2+ ATPase (SERCA) pumps on the SR as a three dimensional random walk through the PM-SR junctional cytoplasmic spaces. Electron microscopic ultrastructural images of the smooth muscle cells were elaborated with software algorithms to produce a very clear and dimensionally accurate picture of the PM-SR junctions. From this study, we conclude that it is plausible and possible for enough Ca2+ to pass through the PM-SR junctions to replete the SR during the regenerative Ca2+ release, which underlies agonist induced asynchronous Ca2+ oscillations in vascular smooth muscle. PMID- 17418404 TI - Complex translocation t(6;9;22)(p21.1;q34;q11) at diagnosis is a therapy resistance index in chronic myeloid leukaemia. PMID- 17418405 TI - Fumarate hydratase deficiency and cancer: activation of hypoxia signaling? AB - Molecular genetic analysis of hereditary leiomyomatosis and renal cell cancer (HLRCC) unexpectedly revealed germline defects in the gene encoding the Krebs cycle enzyme fumarate hydratase (FH), stimulating great interest in the underlying mechanism of oncogenesis. It has been proposed that the associated accumulation of fumarate competitively inhibits the 2-oxoglutarate-dependent dioxygenases that regulate hypoxia-inducible factor (HIF), thus activating oncogenic hypoxia pathways. In this issue of Cancer Cell, Pollard and colleagues describe a genetic mouse model of FH deficiency that recapitulates aspects of the human disease, including HIF activation and renal cysts, enabling further insights into this unusual cancer syndrome. PMID- 17418406 TI - Modeling synovial sarcoma: timing is everything. AB - Synovial sarcoma is characterized by the presence of a fusion protein involving SYT and SSX2. In this issue of Cancer Cell, Haldar et al. have genetically engineered a mouse model of this disease. They show that expression of the SYT SSX2 fusion gene yields a highly penetrant and representative model of human synovial sarcoma, but only if expression occurs in a particular biologic context. The mouse model will be a valuable resource for studying tumor biology but is also a striking example of how important understanding of normal tissue and developmental biology is to our understanding of cancer. PMID- 17418407 TI - Genome-wide SNP analysis in cancer: leukemia shows the way. AB - The application of novel genetic/genomic technologies to the study of acute leukemia has frequently been a proving ground for such approaches in cancer. Recent development of high-resolution single-nucleotide polymorphism (SNP) arrays allows detailed assessment of the genomes in cancer cells. A recent study by Mullighan et al. uses SNP arrays to assess copy number alterations in a large group of childhood acute lymphoblastic leukemias and demonstrates frequent mutation of genes encoding transcription factors important for B cell development. These studies not only provide information about the multistep development of leukemia, but also demonstrate the potential for this approach in other cancers. PMID- 17418408 TI - Targeted inactivation of fh1 causes proliferative renal cyst development and activation of the hypoxia pathway. AB - Germline mutations in the fumarate hydratase (FH) tumor suppressor gene predispose to leiomyomatosis, renal cysts, and renal cell cancer (HLRCC). HLRCC tumors overexpress HIF1alpha and hypoxia pathway genes. We conditionally inactivated mouse Fh1 in the kidney. Fh1 mutants developed multiple clonal renal cysts that overexpressed Hif1alpha and Hif2alpha. Hif targets, such as Glut1 and Vegf, were upregulated. We found that Fh1-deficient murine embryonic stem cells and renal carcinomas from HLRCC showed similar overexpression of HIF and hypoxia pathway components to the mouse cysts. Our data have shown in vivo that pseudohypoxic drive, resulting from HIF1alpha (and HIF2alpha) overexpression, is a direct consequence of Fh1 inactivation. Our mouse may be useful for testing therapeutic interventions that target angiogenesis and HIF-prolyl hydroxylation. PMID- 17418409 TI - Mouse model of human ovarian endometrioid adenocarcinoma based on somatic defects in the Wnt/beta-catenin and PI3K/Pten signaling pathways. AB - One histologic subtype of ovarian carcinoma, ovarian endometrioid adenocarcinoma (OEA), frequently harbors mutations that constitutively activate Wnt/beta-catenin dependent signaling. We now show that defects in the PI3K/Pten and Wnt/beta catenin signaling pathways often occur together in a subset of human OEAs, suggesting their cooperation during OEA pathogenesis. Deregulation of these two pathways in the murine ovarian surface epithelium by conditional inactivation of the Pten and Apc tumor suppressor genes results in the formation of adenocarcinomas morphologically similar to human OEAs with 100% penetrance, short latency, and rapid progression to metastatic disease in upwards of 75% of mice. The biological behavior and gene expression patterns of the murine cancers resemble those of human OEAs with defects in the Wnt/beta-catenin and PI3K/Pten pathways. PMID- 17418410 TI - HIF-2alpha promotes hypoxic cell proliferation by enhancing c-myc transcriptional activity. AB - HIF-2alpha promotes von Hippel-Lindau (VHL)-deficient renal clear cell carcinoma (RCC) tumorigenesis, while HIF-1alpha inhibits RCC growth. As HIF-1alpha antagonizes c-Myc function, we hypothesized that HIF-2alpha might enhance c-Myc activity. We demonstrate here that HIF-2alpha promotes cell-cycle progression in hypoxic RCCs and multiple other cell lines. This correlates with enhanced c-Myc promoter binding, transcriptional effects on both activated and repressed target genes, and interactions with Sp1, Miz1, and Max. Finally, HIF-2alpha augments c Myc transformation of primary mouse embryo fibroblasts (MEFs). Enhanced c-Myc activity likely contributes to HIF-2alpha-mediated neoplastic progression following loss of the VHL tumor suppressor and influences the behavior of hypoxic tumor cells. PMID- 17418411 TI - The differentiation and stress response factor XBP-1 drives multiple myeloma pathogenesis. AB - Multiple myeloma (MM) evolves from a highly prevalent premalignant condition termed MGUS. The factors underlying the malignant transformation of MGUS are unknown. We report a MGUS/MM phenotype in transgenic mice with Emu-directed expression of the XBP-1 spliced isoform (XBP-1s), a factor governing unfolded protein/ER stress response and plasma-cell development. Emu-XBP-1s elicited elevated serum Ig and skin alterations. With age, Emu-xbp-1s transgenics develop features diagnostic of human MM, including bone lytic lesions and subendothelial Ig deposition. Furthermore, transcriptional profiles of Emu-xbp-1s lymphoid and MM cells show aberrant expression of known human MM dysregulated genes. The similarities of this model with the human disease, coupled with documented frequent XBP-1s overexpression in human MM, serve to implicate XBP-1s dysregulation in MM pathogenesis. PMID- 17418412 TI - JUN oncogene amplification and overexpression block adipocytic differentiation in highly aggressive sarcomas. AB - The human oncogene JUN encodes a component of the AP-1 complex and is consequently involved in a wide range of pivotal cellular processes, including cell proliferation, transformation, and apoptosis. Nevertheless, despite extensive analyses of its functions, it has never been directly involved in a human cancer. We demonstrate here that it is highly amplified and overexpressed in undifferentiated and aggressive human sarcomas, which are blocked at an early step of adipocyte differentiation. We confirm by cellular and xenograft mouse models recapitulating these sarcoma genetics that the failure to differentiate is dependent upon JUN amplification/overexpression. PMID- 17418413 TI - A conditional mouse model of synovial sarcoma: insights into a myogenic origin. AB - Synovial sarcoma is an aggressive soft-tissue malignancy marked by a unique t(X;18) translocation leading to expression of a chimeric SYT-SSX fusion protein. We report here a mouse model of synovial sarcoma based on conditional expression of the human SYT-SSX2. Using this model, we have identified myoblasts as a potential source of synovial sarcoma. Remarkably, within the skeletal muscle lineage, while expression of the oncoprotein in immature myoblasts leads to induction of synovial sarcoma with 100% penetrance, its expression in more differentiated cells induces myopathy without tumor induction. We also show that early widespread expression of the fusion protein disrupts normal embryogenesis, causing lethality. PMID- 17418414 TI - A response to Melton based on the best available data. PMID- 17418415 TI - Acute toxicity of oxytetracycline and florfenicol to the microalgae Tetraselmis chuii and to the crustacean Artemia parthenogenetica. AB - Aquaculture systems are a potentially significant source of antibacterial agents to the aquatic environment. The antibacterials oxytetracycline (OTC) and florfenicol (FLO) have been widely used in aquaculture. These pharmaceuticals may cause deleterious effects on wild aquatic organisms accidentally exposed to them. Therefore, the objective of this study was to evaluate the acute toxicity of OTC and FLO to the microalgae Tetraselmis chuii and to the crustacean Artemia parthenogenetica, using culture growth inhibition and death, respectively, as effect criteria. OTC and FLO were found to inhibit the growth of T. chuii cultures, with 96 h IC(50) values of 11.18 and 6.06 mg/L, respectively. OTC 24 and 48 h LC(50) values for A. parthenogenetica were 871 and 806 mg/L, respectively. FLO did not cause mortality of A. parthenogenetica. These results indicate that OTC and FLO are considerably more toxic to T. chui than to A. parthenogenetica. They also indicate that the concentrations required to induce mortality to A. parthenogenetica only in exceptional conditions will occur in the environment. PMID- 17418416 TI - Relevance to self: A brief review and framework of neural systems underlying appraisal. AB - We argue that many similar findings observed in cognitive, affective, and social neuroimaging research may compose larger processes central to generating self relevance. In support of this, recent findings from these research domains were reviewed to identify common systemic activation patterns. Superimposition of these patterns revealed evidence for large-scale supramodal processes, which are argued to mediate appraisal of self-relevant content irrespective of specific stimulus types (e.g. words, pictures) and task domains (e.g. induction of reward, fear, pain, etc.). Furthermore, we distinguish between two top-down sub-systems involved in appraisal of self-relevance, one that orients pre-attentive biasing information (e.g. anticipatory or mnemonic) to salient or explicitly self relevant phenomena, and another that engages introspective processes (e.g. self reflection, evaluation, recollection) either in conjunction with or independent of the former system. Based on aggregate patterns of activation derived from the reviewed studies, processes in a ventral medial prefrontal cortex (MPFC) subcortical network appear to track with the former pathway, and processes in a dorsal MPFC-cortical-subcortical network with the latter. As a whole, the purpose of this framework is to re-conceive the functionality of these systems in terms of supramodal processes that more directly reflect the influences of relevance to the self. PMID- 17418417 TI - Epi-allelic Erk1 and Erk2 knockdown series for quantitative analysis of T cell Erk regulation and IL-2 production. AB - Erk activation is often used as a downstream pathway indicator of TCR signaling, generally in terms of both Erk1 and Erk2 isoforms measured together. In order to investigate potential distinctions between Erk1 and Erk2 regulation and effects downstream of TCR ligation, we generated a series of stable and independent Erk1 and Erk2 shRNA knockdown lines in the 1B6 T cell hybridoma. We observed no compensatory effect by opposite isoform upregulation, and found similar fractions of total phosphorylated Erk1/2 across this epi-allelic series in response to both anti-CD3 and peptide-MHC stimulation of TCR. Moreover, a previous prediction of an isoform-independent linear relationship between Erk activation and IL-2 production was confirmed. The effect of the shRNA-mediated knockdowns in reducing IL-2 production was observed to be stronger than that arising from pharmacological MEK inhibition at comparable degrees of ERK1/2 phosphorylation levels. PMID- 17418418 TI - Archipelago keratitis: a clinical variant of recurrent herpetic keratitis? AB - OBJECTIVE: To describe archipelago keratitis, a presumed clinical variant of herpetic epithelial keratitis. DESIGN: Case series. PARTICIPANTS: A series of 6 patients with an unusual form of superficial keratitis. METHODS: History, including age, gender, clinical evolution, and treatment; slit-lamp biomicroscopy findings; in vivo confocal microscopy findings; and corneal epithelial scrapings were analyzed. MAIN OUTCOME MEASURES: Clinical ocular examination, a diagnostic workup including corneal scraping for herpesvirus polymerase chain reaction, in vivo confocal microscopy, and therapeutic outcome. RESULTS: The authors describe a series of 6 patients with keratitis consisting of foci of epithelial erosions associated with subepithelial nummular inflammatory infiltrates and disposed in a radial, centripetal, archipelagolike pattern originating from the limbus. All the patients had a past history of herpetic epithelial keratitis, herpetic vesicles on the ipsilateral lid, or both. Polymerase chain reaction-based screening for herpes simplex virus 1 and 2 in corneal scrapings demonstrated positive results in 2 patients. In vivo corneal confocal microscopy revealed focal areas of hyperreflective epithelial cells and hyperreflective subepithelial dendritic structures overlying activated keratocytes. All the patients improved with oral valacyclovir treatment followed by topical steroid therapy. CONCLUSIONS: Archipelago keratitis may be a new clinical variant of herpetic keratitis, reflecting herpetic dissemination from the limbus to the center of the cornea. PMID- 17418419 TI - Propionibacterium acnes: an agent of prosthetic joint infection and colonization. AB - BACKGROUND: Prosthetic joint infection (PJI) can present a diagnostic challenge, especially with slow-growing and poorly virulent bacteria. OBJECTIVE: To describe the epidemiological, clinical and biological characteristics of Propionibacterium acnes PJI, their treatments and outcomes and compare 2 clinical pictures (according to the time PJI symptoms appeared after the index operation: < or = 2 years, > 2 years). METHODS: We conducted a cohort study on P. acnes PJI. Diagnosis was based on > or = 2 positive cultures of intraoperative specimens taken during revision arthroplasties for infection or presumed aseptic loosening. RESULTS: Fifty patients with prosthetic hip (34), knee (10) or shoulder (6) infections were included and analyzed according to their symptom-free interval: < or = 2 years for 35 and > 2 years for 15 (mean interval: 11+/-6 years). The numbers of previous prostheses (p=0.04) were higher for the shorter-interval group, which had more frequent signs of infection (p=0.004). These findings suggest infection in most of the patients whose PJI symptoms appeared: < or = 2 years after the index operation, and colonization in the majority of those whose symptoms appeared > 2 years after index surgery. Treatment combining exchange arthroplasty with prolonged intravenous antibiotics was successful for 92% of the patients. CONCLUSION: P. acnes can cause different types of PJI: late chronic infections, colonization of loosened prostheses and, exceptionally, acute postoperative infections. PMID- 17418420 TI - VZV infection in pregnancy: a retrospective review over 5 years in Sheffield and discussion on the potential utilisation of varicella vaccine in prevention. AB - OBJECTIVES: To study retrospectively the epidemiology, demography and clinical issues related to varicella in pregnancy in a UK city over a 5-year period and help inform the debate on the potential of varicella immunisation in prevention. METHODS: The hospital records of pregnant women with varicella receiving care at the Regional Department of Infection and Tropical Medicine in Sheffield between 1997 and 2002 were reviewed. Data on pregnant women with varicella not presenting acutely to medical care were obtained. The use of Varicella Zoster Immune Globulin (VZIG) in prevention of varicella during the same 5-year period was determined. The records from the maternity department of Sheffield Hospitals for women undergoing VZV antibody testing between January and December 2004 were reviewed. Data on annual number of deliveries were recorded and the neonatology database used as a source of information regarding effects of chickenpox on the baby. RESULTS: The incidence of varicella infection in pregnancy was at least 6 per 10,000 deliveries. Nineteen pregnant women with varicella were admitted to hospital. Three had pneumonia. Infection occurred in the first pregnancy in a quarter of cases. The minimum cost for all cases admitted to hospital during this period (basic costs per day) was 20,520 pounds sterling. The cost of VZIG use for chickenpox during the same period adjusted for the population size was 10,881 pounds sterling. This was not a comprehensive health economic study and did not attempt to assess additional GP, midwifery, obstetric or social costs nor costs associated with those who did not attend hospital. Two hundred and thirty-three women underwent VZV antibody test during 2004 usually after contact with chickenpox. Sixty percent of women in contact with chickenpox did not present to their GP or hospital immediately. CONCLUSION: Varicella in pregnancy may be associated with significant morbidity and health care cost and prevention by immunisation is desirable. Though targeted vaccination is attractive, screening in pregnancy followed by a post-partum varicella immunisation programme would fail to protect 25% and would be associated with logistical challenges not occurring with rubella immunisation. Varicella is now a preventable disease by immunisation. Exposure in pregnancy with or without infection has financial costs related to antibody testing and prophylaxis. Infection in pregnancy may be associated with additional costs and potential morbidity to mother and baby. Potential immunisation strategies are considered. PMID- 17418421 TI - Yawning frequency and distribution in preterm and near term infants assessed throughout 24-h recordings. AB - Yawning has been observed in foetuses and preterm infants. The aim of this study was to assess the frequency and the 24 h distribution of yawning in preterm infants. Twelve low-risk infants between 31 and 40 weeks of post-conceptional age (PCA) were continuously video-recorded for 24 h in their incubator. Spontaneous yawning was defined as opening of the mouth to its full extension in a dramatic stretch movement. The results showed that the rate of yawning across the 24-h period was 1.10/h. The highest incidence of yawns was in the waking motility pattern when compared to active sleep or quiet sleep motility patterns. Between 31 and 40 weeks, yawn incidence significantly decreased mainly during the day. The marked decrease in yawn frequency with age may be related to the development of circadian and homeostatic control of sleep and wake. PMID- 17418422 TI - A feed-forward neural logic based on synaptic and volume transmission. AB - We consider a homeostatic mechanism to maintain a plastic layer of a feed-forward neural network reactive to a long sequence of signals, with neither falling in a fixed point of the state space nor undergoing in overfitting. Homeostasis is achieved without asking the neural network to be able to pursue an offset through local feedbacks. Rather, each neuron evolves monotonically in the direction increasing its own parameter, while a global feedback emerges from volume transmission of a homostatic signal. Namely: 1) each neuron is triggered to increase its own parameter in order to exceed the mean value of all of the other neurons' parameters, and 2) a global feedback on the population emerges from the composition of the single neurons behavior paired with a reasonable rule through which surrounding neurons in the same layer are activated. We provide a formal description of the model that we implement in an ad hoc version of pi-calculus. Some numerical simulations will depict some typical behaviors that seem to show a plausible biological interpretation. PMID- 17418423 TI - Antiphospholipid antibodies and pregnancy loss: a disorder of inflammation. AB - The antiphospholipid syndrome (APS) is a leading cause of miscarriage and maternal and fetal morbidity. APS is characterized by thrombosis and pregnancy loss that occur in the presence of antiphospholipid (aPL) antibodies. Using a mouse model of APS induced by passive transfer of human aPL antibodies, we have shown that complement activation plays an essential and causative role in pregnancy loss and fetal growth restriction, and that blocking activation of the complement cascade rescues pregnancies. Conventional treatment for APS patients is sub-anticoagulant doses of heparin throughout pregnancy. Could heparin prevent pregnancy loss by inhibiting complement? In our experimental model of APS, heparin inhibits activation of complement on trophoblasts in vivo and in vitro, and anticoagulation in and of itself is not sufficient to prevent pregnancy complications. These studies underscore the importance of inflammation in fetal injury associated with aPL antibodies and raise the importance of developing and testing targeted complement inhibitory therapy for patients with APS. PMID- 17418424 TI - Malignant orofacial neoplasms in children and adolescents: a clinicopathologic review of cases in a Nigerian tertiary hospital. AB - OBJECTIVE: The aim of the study was to determine the relative frequency of orofacial malignant neoplasm in children and adolescents. METHODS: A retrospective review of malignant orofacial tumours in children and adolescents 1000 neurons in a wide area, (ii) single-cell resolution, (iii) identifiable location of neurons and (iv) detection of non active neurons during the observation period. We review herein the principle, history, utility and limitations of fMCI. PMID- 17418440 TI - Normal mortality in the elderly with diabetes under strict glycemic and blood pressure control: outcome of 6-year prospective study. AB - Mortality, macroangiopathic events and end-stage renal disease (ESRD) in the elderly under long-term, intensive multifactorial diabetes control were prospectively investigated. Three hundred and eighty-eight elderly patients (> or =65 years) with type 2 diabetes (the mean age 72.9 years, men/women ratio 176/212) were followed-up for 6 years with HbA1c 7.0%, BP 145/80 mmHg and total cholesterol<240 mg/dl as targets. The mean baseline HbA1c was 6.8%, BP 137/74 mmHg and total cholesterol 196 mg/dl, and corresponding values upon closing 6.9%, 134/72 mmHg and 188 mg/dl respectively. Mortality rate was 19.6%/6 years (1.01 times that of age- and sex-matched general population), and macroangiopathic events developed in 142 (36.6%) and ESRD in 9 (2.3%). Independent risk factors: low glomerular filtration rate (GFR) (P<0.001), prior stroke (P=0.002), age (P=0.001) and DeltaBMI (P=0.001) for mortality; prior stroke (P<0.001) and coronary events (P=0.042), high LDL-cholesterol (P=0.004), low GFR (P=0.028), and past maximum BMI (P=0.032) and age (P=0.019) for macroangiopathy; low GFR (P<0.001) for ESRD. No smoking was an independent protective factor for mortality (P=0.008). In conclusion, normal mortality was attained in the elderly under intensive mutifactorial diabetes control. Renal dysfunction, prior stroke, high LDL-cholesterol, and prior obesity were prominent risks for mortality, macroangiopathy and/or ESRD. PMID- 17418441 TI - Ain't misbehavin' ? Genotype-environment interactions and the genetics of behavior. AB - Although the multiple interacting genes affecting complex traits can readily be dissected, how much genotype-environment interactions contribute to variation in complex traits remains elusive. A recent study that quantified several behavioral phenotypes on the same mouse strains in different laboratories decades apart, shows that some behavioral differences between laboratories remain greatly replicable, whereas others are less robust over time. This report, together with studies from Drosophila, stresses the importance of understanding genotype environment interactions. PMID- 17418442 TI - Adaptation or biased gene conversion? Extending the null hypothesis of molecular evolution. AB - The analysis of evolutionary rates is a popular approach to characterizing the effect of natural selection at the molecular level. Sequences contributing to species adaptation are expected to evolve faster than nonfunctional sequences because favourable mutations have a higher fixation probability than neutral ones. Such an accelerated rate of evolution might be due to factors other than natural selection, in particular GC-biased gene conversion. This is true of neutral sequences, but also of constrained sequences, which can be illustrated using the mouse Fxy gene. Several criteria can discriminate between the natural selection and biased gene conversion models. These criteria suggest that the recently reported human accelerated regions are most likely the result of biased gene conversion. We argue that these regions, far from contributing to human adaptation, might represent the Achilles' heel of our genome. PMID- 17418443 TI - More is not always better: the genetic constraints of polyploidy. AB - Polyploid cells are a characteristic feature of certain human tissues, and notably many cancers. In a systematic genomic screen in yeast, Storchova and co workers identified the genetic requirements of tetraploidy. Surprisingly, they showed that only three connected pathways are essential for the viability of tetraploid yeast cells. These data provide exciting new targets that might be essential specifically in polyploid cancer cells. PMID- 17418444 TI - When gene marriages don't work out: divorce by subfunctionalization. AB - We describe how a bifunctional gene, encoding two proteins by alternative splicing, arose when the chloroplast gene RPL32 integrated into an intron of the nuclear gene SODcp in an ancestor of mangrove and poplar trees. Mangrove retains the alternatively spliced chimeric gene, but in poplar it underwent duplication and complete subfunctionalization, through complementary structural degeneration, to re-form separate RPL32 and SODcp genes. PMID- 17418445 TI - Positive and negative selection on the mitochondrial genome. AB - Several recent studies have confirmed that mitochondrial DNA variation and evolution are not consistent with the neutral theory of molecular evolution and might be inappropriate for estimating effective population sizes. Evidence for the action of both positive and negative selection on mitochondrial genes has been put forward, and the complex genetics of mitochondrial DNA adds to the challenge of resolving this debate. The solution could lie in distinguishing genetic drift from 'genetic draft' and in dissecting the physiology of mitochondrial fitness. PMID- 17418446 TI - Identification and validation of S100A7 associated with lung squamous cell carcinoma metastasis to brain. AB - To identify potential markers associated with non-small cell lung cancer (NSCLC) metastasis to brain, comparative proteome analysis on two lung squamous cell carcinoma (SCC) cell lines, NCI-H226 and H226Br (the brain metastatic cell line of NCI-H226), was performed using two-dimensional electrophoresis (2-DE) followed by a tandem mass spectrometer with a matrix-assisted laser desorption/ionization (MALDI) source. Twenty differential proteins were identified, of which 6 proteins were up-regulated in H226Br cell compared with NCI-H226 cells, whereas 14 proteins were down-regulated. S100A7 and 14-3-3sigma, two of candidate proteins significantly upregulated and downregulated in H226Br cell, were selected to verify the liability of the differential proteins by Western blot. The results were in accordance with 2-D data. To determine whether S100A7 overexpression is actually associated with SCC metastasis to brain, S100A7 protein was testified in 10 brain metastasis tissues from NSCLC, 38 primary NSCLC tissues including half matched local positive lymph nodes, 5 primary brain tumors and 2 non-cancer brain tissues by immunohistochemistry. Of particular interest to us was that the positive staining of S100A7 could be found in 3/5 (60%) brain metastases tissue from SCC and 8/21 (38%) the primary lung SCC tissues, while no positive staining was observed in the brain metastases tissue from Ad (n=5), the primary adenocarcinoma (Ad) tissues (n=17), the primary brain tumors (n=5), all local positive lymph nodes from the primary NSCLC (n=19) and non-cancer brain tissues (n=2). These findings suggest that S100A7 expression is closely associated with SCC metastasis to brain and may be a potential biomarker for monitoring the development of SCC. PMID- 17418447 TI - Syndrome of inappropriate antidiuretic hormone secretion and ectopic ACTH production in small cell lung carcinoma. AB - BACKGROUND: Small cell lung carcinoma (SCLC) is frequently associated with a paraneoplastic syndrome. CASE REPORT: We report on a rare case of SCLC presenting with syndrome of inappropriate antidiuretic hormone (SIADH) secretion and ectopic ACTH production. In accordance with the endocrine evaluation, immunohistochemical staining of the tumour was positive for ADH and ACTH. Chemotherapy with etoposide and carboplatin induced a nearly complete remission. CONCLUSION: Simultaneous secretion of ADH and ACTH is infrequently reported with only five cases described in the literature. SIADH in patients with ectopic ACTH syndrome may be underdiagnosed due to the antagonistic hormone actions of cortisol and ADH on renal sodium excretion. PMID- 17418448 TI - The evolution of sex ratios and sex-determining systems. AB - Sex determination is a fundamental process governed by diverse mechanisms. Sex ratio selection is commonly implicated in the evolution of sex-determining systems, although formal models are rare. Here, we argue that, although sex ratio selection can induce shifts in sex determination, genomic conflicts between parents and offspring can explain why single-factor systems (e.g. XY/XX or ZW/ZZ) are common even in species that experience selection for biased sex ratios. Importantly, evolutionary shifts in sex determination do not always result in the biased production of sons and daughters sensu sex ratio theory. Thus, equal sex ratios might be an emergent character of sex-determining systems even when biased sex ratios are favored by selection. PMID- 17418449 TI - Patient-tailored acetylcysteine administration. PMID- 17418450 TI - Toxicity from the use of niacin to beat urine drug screening. AB - Niacin (vitamin B3) is promoted for rapidly clearing the body of drugs of abuse, such as cocaine and cannabis, and is alleged to interfere with urine drug screening. We present 4 cases of such novel use associated with significant adverse effects. Two cases had isolated skin manifestations, whereas the other 2 presented with life-threatening manifestations, including nausea, vomiting, dizziness, hepatotoxicity, metabolic acidosis, and hypoglycemia evolving into hyperglycemia. One patient also had profound neutrophilia and QT(C)-interval prolongation. All patients improved after cessation of the drug use and supportive treatment. Health care providers should be aware of these potential adverse effects of niacin and of the misguided use of this vitamin by patients seeking to interfere with urine drug screening. PMID- 17418451 TI - Staurosporine differentiation of NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells induces selectivity of NPFF activity towards opioid receptors. AB - Activation of the NPFF(2) receptor reduces the inhibitory effect of opioids on the N-type Ca(2+) channel. Although this anti-opioid effect is specific for opioid receptors in neurons and tissues, it also affects NPY Y2 and alpha(2) adrenoreceptors in undifferentiated SH-SY5Y cells stably expressing the NPFF(2) receptor. To test whether this difference could be due to the immaturity of these cells, they were differentiated to a noradrenergic neuronal phenotype with staurosporine. The differentiated cells ceased to divide and grew long, thin neurites. The inhibition of the depolarization-triggered Ca(2+) transient by activation of G(i)-coupled receptors was either unaffected (micro-opioid), increased (NPY), reduced (NPFF(2)) or lost (alpha(2)-adrenoreceptors). Following a 20 min incubation with 1DMe, the effect of DAMGO was reduced, as in undifferentiated cells, but the effect of NPY was no longer affected. Staurosporine differentiation did not modify the coupling of the micro-opioid and NPFF(2) receptors to the G(i/o) proteins. We suggest that the specificity of the effect of NPFF may not reside in the molecular mechanism of its anti-opioid activity itself but in the organization of receptors within the membrane. PMID- 17418452 TI - Synthesis and CNS depressant activity of some novel 3-[5-substituted 1,3,4 thiadiazole-2-yl]-2-styryl quinazoline-4(3H)-ones. AB - A series of novel 3-[5-substituted phenyl-1,3,4-thiadiazole-2-yl]-2-styryl quinazoline-4(3H)-ones were synthesized and evaluated for anticonvulsant, sedative-hypnotic and CNS depressant activities. After i.p. injection to mice at doses of 30, 100, and 300mg/kg body weight 2-styrylquinazolin-4(3H)-one derivatives were examined in the maximal electroshock induced seizures (MES) and subcutaneous pentylenetetrazole (scPTZ) induced seizure models in mice. The neurotoxicity was assessed using the rotorod method. Out of eighteen compounds only 4a, 4d, 4e, 4j and 4k showed anticonvulsant activity in one or more test models. All except 4e and 4f exhibited significant sedative-hypnotic activity via actophotometer screen. CNS depressant activity screened with the help of the forced swim pool method resulted into some potent compounds. From the experimental observation it can be concluded that synthesized compounds exhibited relatively better sedative-hypnotic and CNS depressant activities. PMID- 17418453 TI - Novel lbeta-methylcarbapenems having cyclic sulfonamide moieties: synthesis and evaluation of in vitro antibacterial activity. AB - The synthesis of a new series of 1beta-methylcarbapenems having cyclic sulfonamide moieties is described. Their in vitro antibacterial activities against both Gram-positive and Gram-negative bacteria were tested and the effect of substituent on the pyrrolidine ring was investigated. A particular compound (IIIi) having 2-methyl-[1,2,6]thiadiazinan-1,1-dioxide moiety showed the most potent antibacterial activity. PMID- 17418454 TI - Synthesis of some novel heterocyclic compounds derived from diflunisal hydrazide as potential anti-infective and anti-inflammatory agents. AB - Three novel series of 2',4'-difluoro-4-hydroxybiphenyl-3-carboxylic acid derivatives namely 4-substituted-1,2,4-triazoline-3-thiones (4a-g); 2-substituted 1,3,4-thiadiazoles (5a-g) and 2-substituted-1,3,4-oxadiazoles (6a-g) have been synthesized. Twenty-one of the newly synthesized compounds were tested against various bacteria, fungi, yeast species and virus. In addition, we have replaced the carboxylic acid group of diflunisal with heterocycles and the anti inflammatory activity of heterocycles reported here. Compound (5d) showed activity against Escherichia coli A1 and Streptococcus pyogenes ATCC-176 at a concentration of 31.25 microg/mL, whereas cefepime, the drug used as standard, has been found less active against the bacteria mentioned above. Compound (4b) has exhibited activity against Aspergillus variecolor and Trichophyton rubrum at a concentration of 31.25 and 15.25 microg/mL, whereas Amphotericin B, the drug used as standard, has been found less active against the yeast and fungi. The highest antiviral activity was found in the 1,3,4-thiadiazole derivative (5a) having a methyl group at 2nd position against Sindbis virus at 9.6 microg/mL. Compound (4c) exhibited the highest anti-inflammatory activity (73.03%) whereas diflunisal, the drug used as standard, has been found less active (24.16%). Compound (5f) presented similar antinociceptive activity with the standard drug (paw withdrawal latency was 19.21 s compared to that of diflunisal which was 19.14s, in hot plate test). PMID- 17418455 TI - Evaluation of antitumor immunity efficacy of epitope-based vaccine with B16 cell line coexpressing HLA-A2/H-2kb and CTL multiepitope in HLA transgenic mice. AB - Epitope-based vaccination strategies designed to induce tumor-specific CD8 CTL are being widely considered for cancer immunotherapy. HLA-A2-transgenic mouse is a useful tool for measuring the CTL responses in vitro. However, tumor vaccine development is required to address the variables that are not easily evaluated by in vitro assays. With the objective of extending the usage of A2-tansgenic mouse in vaccine efficacy assay, here, we established a B16 tumor cell line coexpressing HLA-A*0201/H-2Kb chimeric gene and a polyepitope construct based on the use of a mammalian expression vector pIRES. The value as a tool for evaluating the antitumor efficacy in vitro as well as in experimental tumor challenge model in vivo has been tested. We found that priming with the polyepitope construct and boosting with the mixture of peptide in A2-transgenic mice resulted in: (1) CTL responses not only against the peptide-sensitized T2 and SW480 cell lines but also the non-sensitized reconstructed B16 cell line; (2) expression of HLA-A*0201/H-2Kb chimeric gene and polyepitopes by B16 led to its rejection by immunized A2-transgenic mice. These data established that the reconstructed B16 cell line stably expressed and efficiently presented the HCC derived CTL epitopes, making B16 based melanoma suitable for the evaluation of the antitumor efficacy of immune responses to these epitopes. Collectively, these data indicate that the use of this method allows for directly testing of HLA-A2 restricted epitope immunogenicity in the A2-transgenic mice. PMID- 17418456 TI - Immune responses of pigs inoculated with a recombinant fowlpox virus coexpressing GP5/GP3 of porcine reproductive and respiratory syndrome virus and swine IL-18. AB - Two recombinant fowlpox viruses (rFPV-ORF5-ORF3 and rFPV-IL-18-ORF5-ORF3) containing the ORF5/ORF3 cDNAs of PRRSV (strain Chang Chun) and IL-18 of swine were constructed and evaluated for theirs abilities to induce humoral and cellular responses in piglets. In addition, their abilities to protect piglets against homologous virus challenge were examined. All piglets were given booster vaccinations at 21 days after the initial inoculation, and all piglets were challenged at 60 after the initial inoculation. Control groups were inoculated with wild-type fowlpox virus (wtFPV). All animals vaccinated with rFPV-ORF5-ORF3 and rFPV-IL-18-ORF5-ORF3 developed specific anti-PRRSV ELISA antibody and neutralizing antibody, as well as T-lymphocyte proliferation response. To evaluate the cellular immune function, IFN-gamma production in pigs serum and T lymphocytes (CD4 and CD8 T cells) in peripheral blood were examined. Following challenge with a pathogenic strain of PRRSV (strain Chang Chun), piglets inoculated with recombinant fowlpox virus (rFPV) showed lower (P<0.05) temperature, viremia and virus load in bronchial lymph nodes than control animals, suggesting the establishment of partial protection against PRRSV infection. The results demonstrated the potential use of a fowlpox virus-based recombinant vaccine in the control and prevention of PRRSV infections. PMID- 17418457 TI - Evaluation of protective effect of recombinant dense granule antigens GRA2 and GRA6 formulated in monophosphoryl lipid A (MPL) adjuvant against Toxoplasma chronic infection in mice. AB - To investigate the vaccine potential of both the Toxoplasma GRA2 and GRA6 antigens, the full length recombinant proteins were produced in Escherichia coli, formulated in MPL adjuvant, and used alone and in combination ("mix"), to immunize CBA/J mice. Although high ratios of specific IgG2a/IgG1 were measured against both proteins, only spleen cells from GRA2-immunized mice and mix immunized mice produced high amounts of both IFN-gamma and IL-2 upon induction with Toxoplasma gondii Excretory-Secretory Antigens. Intra peritoneal challenge with Toxoplasma cysts resulted in significant reduction of brain cysts in GRA2- and in mix-vaccinated mice only. This study shows the protective efficacy of recombinant GRA2 against chronic infection by T. gondii and confirms the utility of MPL adjuvant in enabling a vaccine candidate to induce a protective Th1 immune response. PMID- 17418458 TI - Advocating rabies control in Asia. PMID- 17418459 TI - DNA vaccination against the parasite enzyme gamma-glutamylcysteine synthetase confers protection against Leishmania donovani infection. AB - In this study the potential of using Leishmania donovani gamma-glutamylcysteine synthetase (glutamate-cysteine ligase, gamma-GCS) as a rational target for vaccine development was determined. Mice, immunised with plasmid containing the full gene sequence for gamma-GCS (pVAXgammaGCS) or plasmid alone (pVAX control), were challenged with a high dose of L. donovani amastigotes to give a stringent test of the ability of the vaccine to protect against infection. Vaccination with pVAXgammaGCS resulted in the production of specific IgG1 and IgG2a antibodies and resulted in significantly lower liver parasite burdens compared to controls. Protection was also associated with a significant increase in cell-mediated immunity, demonstrated as an increase in nitrite production by ConA stimulated splenocytes, an increase in the percentage of splenic CD3+CD4+ cells, and enhanced granuloma maturation, compared to control values. PMID- 17418460 TI - The use of vaccination to combat multiple introductions of Notifiable Avian Influenza viruses of the H5 and H7 subtypes between 2000 and 2006 in Italy. AB - Since 1999, Italy has been challenged by several epidemics of Notifiable Avian Influenza (NAI) of the H5 and H7 subtypes, occurring in the densely populated poultry areas of northern part of the country. Vaccination with a conventional vaccine containing a seed strain with a different neuraminidase subtype to the field virus was used to complement biosecurity and restriction measures as part of an overall eradication strategy. This vaccination technique, known as the "DIVA-Differentiating Infected from Vaccinated Animals" system, enabled, the identification of field exposed flocks and ultimately the eradication of H7N1, H7N3 and H5N2 infections. A bivalent H5/H7 prophylactic vaccination programme of defined poultry populations was introduced subsequently to increase their resistance to field infection. Retrospective analysis of the outbreaks identified important reservoir species such as quail, and demonstrated clearly the higher susceptibility of turkeys to infection. Data generated during 6 years of experience with vaccination against Avian Influenza (AI) indicate that it is a useful tool to limit secondary spread and possibly prevent the introduction of AI viruses in a susceptible population. The Italian AI control programme including vaccination was managed in a flexible manner and enabled the continuation of international trade. It is imperative that if vaccination is to be used to combat the current H5N1 epidemic it is used in conjunction with other measures and under official supervision. An extraordinary effort is required from international organisations to accredit control strategies so that harmonised and validated programs can be implemented. Transparency and sharing of field results from countries that are practising such programmes is crucial to the progressive control and ultimately the eradication of NAI infections in the animal reservoir. PMID- 17418461 TI - Yield increases in intact influenza vaccine virus from chicken allantoic fluid through isolation from insoluble allantoic debris. AB - A yield enhancement technology for use in influenza vaccine manufacturing has been developed to maximize the recovery of influenza virus from allantoic fluid of virus-infected chick embryos; the standard raw material for influenza vaccine. Virus associated with amorphous debris in the allantoic fluid can be dissociated from the debris and recovered, thereby increasing viral yield. Dissociation can be achieved by subjecting the virus-debris complex to conditions of increased ionic strength at defined pH. Multifold increases in viral yield per ml of allantoic fluid were observed. The degree of yield enhancement is strain specific, however, increases were observed in all type A and type B influenza strains tested. The heightened influenza virus recoveries can facilitate rapid vaccine manufacture, with increased numbers of doses produced, and may become essential at a time of influenza pandemic. PMID- 17418462 TI - Radioactivity in the environment (including the Arctic and Antarctic). The International Conference in Nice, France, 2-6 October 2005. PMID- 17418463 TI - Conservative, non-surgical management of lower third molars [Br J Or Maxillofac Surg 2006;44:347-350]. PMID- 17418464 TI - A systematic review of education and evidence-based practice interventions with health professionals and breast feeding counsellors on duration of breast feeding. AB - OBJECTIVE: to examine the effects of training, education and practice change interventions with health professionals and lay breast feeding educator/counsellors on duration of breast feeding. REVIEW METHODS: this was part of a series of reviews of interventions that affect duration of breast feeding. Full details of methods used, including search strategy, are reported separately. SELECTION CRITERIA FOR INCLUDED STUDIES: randomised controlled trials, non randomised controlled trials with concurrent controls and before after studies (cohort or cross-sectional), undertaken in a developed country, published between 1980 and 2003 in any language. The primary outcome was duration of breast feeding. Secondary and process outcomes, including attitude, knowledge and behaviour change of participants, were included from papers that also reported breast feeding duration outcomes. STUDY-QUALITY ASSESSMENT: inclusion and exclusion criteria were applied, data extracted and study quality assessments made by one reviewer and independently checked by another, with a third reviewer to resolve differences, as recommended by the NHS Centre for Reviews and Dissemination's guidance for reviews. FINDINGS: the search identified nine papers. All were before after studies that included the education of health professionals; no studies were identified that related to breast feeding counsellors. In six of the studies, the participants were working with mothers and babies in hospitals (three in the UK, two in Italy and one in France); in three studies, the participants were working in community settings (Canada, Spain and the USA). Two UK studies and two non-UK studies (Spain and USA) involved mothers living in disadvantaged areas. Most interventions aimed to increase knowledge and change professional practice in support of breast feeding. KEY CONCLUSIONS: many of the studies reviewed have methodological limitations. Study settings and contexts vary and lack comparability. Evidence from these studies was insufficient to draw conclusions about overall benefit or harm associated with the interventions. From the studies identified, there seems to be no single way that consistently achieves changes in breast feeding duration. From one of the methodologically more robust studies, it seems that UNICEF/WHO Baby Friendly Hospital Initiative (BFI) training might have the potential to influence breast feeding duration. RECOMMENDATIONS FOR FURTHER RESEARCH: further testing of the BFI initiative within a controlled design. Future research into the education of health-care professionals that relates to the support of breast feeding women should have appropriate theoretical underpinning, describe educational programmes and the context of care delivery comprehensively and use standardised time points in the assessment of the effect of interventions. Intermediate outcomes should also be reported, including those related to the effect on education and practice. PMID- 17418465 TI - Twenty questions on carpal instability. PMID- 17418466 TI - The groundwater contribution to surface water contamination in a region with intensive agricultural land use (Noord-Brabant, The Netherlands). AB - Traditionally, monitoring of soil, groundwater and surface water quality is coordinated by different authorities in the Netherlands. Nowadays, the European Water Framework Directive (EU, 2,000) stimulates an integrated approach of the complete soil-groundwater-surface water system. Based on water quality data from several test catchments, we propose a conceptual model stating that stream water quality at different discharges is the result of different mixing ratios of groundwater from different depths. This concept is used for a regional study of the groundwater contribution to surface water contamination in the Dutch province of Noord-Brabant, using the large amount of available data from the regional monitoring networks. The results show that groundwater is a dominant source of surface water contamination. The poor chemical condition of upper and shallow groundwater leads to exceedance of the quality standards in receiving surface waters, especially during quick flow periods. PMID- 17418467 TI - Impacts of boat paint chips on the distribution and availability of copper in an English ria. AB - Discarded paint chips collected from a leisure boat maintenance facility on the Kingsbridge estuary, SW England, have been fractionated to <63 microm and chemically characterised. At about 16% by weight, Cu was the most abundant metallic component, reflecting its biocidal application in antifouling paint. Bioavailability of Cu in the chips, determined by protein digestion, was about 4%, and sea water leachability was about 8%. Copper concentrations in fractionated intertidal sediment from the estuary were highly variable (<10-460 microg g(-1)). Specifically, greatest concentrations and greatest variability among replicates were found in samples collected near boat maintenance facilities. Bioavailability of Cu in sediment averaged 7% but was also variable. We attribute Cu "hot spots" to heterogeneous contamination of local sediment by small quantities of paint chips. Contamination may arise directly, from relatively inert particulates, or indirectly, via release of Cu from chips to interstitial waters and its subsequent adsorption to local sediment. PMID- 17418468 TI - Perception of place and health: differences between neighbourhoods in the Quebec City region. AB - This paper addresses two questions: (1) Can people's perceptions of problems and social cohesion in the neighbourhood be considered as contextual variables; and (2) are these perceptions related to people's health? Data come from a general health survey carried out in 2004 among 1634 individuals living in three localities of the region of Quebec City, namely a downtown, a suburban and a rural area, which were further subdivided into 34 smaller spatial units, hereafter called neighbourhoods. The survey included questions on individuals' perception of problems (social and environmental) and social cohesion (attraction to neighbourhood, neighbouring and psychological sense of community) in the neighbourhood, as well as questions on self-rated health, long-term disability and self-mastery. A first set of logistic multilevel models was performed to ascertain the existence of neighbourhood variations in the perception of problems and social cohesion, after accounting for individual attributes. A second set of multilevel models was carried out to examine the association between perceived problems and social cohesion in the neighbourhood and people's health. Results show that, after accounting for individual attributes, the perception of problems and social cohesion varies significantly by neighbourhood and/or localities and can be considered as contextual variables. Furthermore, these perceptions of place appear to be significant predictors of people's health. PMID- 17418469 TI - Genetics, insurance and participation: how a Citizens' Jury reached its verdict. AB - Access to life insurance is subject to health discrimination. Whether this discrimination should take into account the results of predictive genetic tests is a topic of public debate. This paper draws from the deliberations of the Edinburgh (Scotland) Citizens' Jury on Genetic Test Results and Life Insurance to evaluate the capacity of one participatory research method to inform debates on genetics and insurance. We show that through a process of knowledge-building, cross-examination and deliberation, 'lay' jurors are able to assimilate complex information, engage in subtle argument and arrive at well-reasoned, clearly warranted conclusions. The Citizens' Jury approach has a further key advantage: it embraces the formation and articulation of normative ideas. It brings public understandings of how things ought to be into an arena dominated by 'expert' opinion. These normative indicators, which in this case relate to fairness and trust, are often overlooked by policy makers. However, they are key to democratic decision-taking and relevant for health promotion. PMID- 17418470 TI - Who kicks the habit and how they do it: socioeconomic differences across methods of quitting smoking in the USA. AB - Although the prevalence of smoking has declined among US adults, an estimated 22.5% of the adult population (45.8 million adults) regularly smoked in 2002. Starting from this level, it will not be possible to achieve the Healthy People national health objectives of a reduction in the prevalence of smoking among adults to less than 12% by 2010 unless the rate of smoking cessation substantially rises from its current average of about 2.5%/year. To achieve that goal it is imperative that we better understand what factors are associated with successful quitting so that policies and resources can be better targeted. We describe the socioeconomic characteristics of smokers who attempt to and successfully quit and show how those characteristics differ across three methods they use in their cessation behavior. The results highlight socioeconomic differences across the methods smokers use and provide evidence that can be used to better target smoking cessation information and resources to smokers most likely to use particular methods. Better targeting is likely to lead to more quits. While it is unlikely that cessation rates can be raised by enough to achieve the reduction in national smoking prevalence that the Healthy People initiative has set, a better understanding of who chooses which method will move us closer to that goal. PMID- 17418471 TI - Evaluation of the potential genotoxicity of chromium picolinate in mammalian cells in vivo and in vitro. AB - Chromium picolinate (CrPic) is a synthetic nutritional supplement primarily used for weight loss and muscle building. Recent studies have indicated that CrPic might be genotoxic and these findings together with the wide-spread consumer use, have increased the concern about its safety. In the present study we investigated the potential genotoxicity of CrPic in mice given a single intraperitoneal injection (up to 3 mg/kgb.wt.) by evaluating the frequency of micronucleated polychromatic erythrocytes (fMNPCE) in peripheral blood, and DNA damage in lymphocytes and hepatocytes. The fMNPCE was evaluated after 42 h and DNA damage after 16 h. Using the Comet assay DNA damage was also monitored in extended-term cultures of human lymphocytes and in L5178Y mouse lymphoma cells that had been exposed for 3h to 500 microM CrPic under different exposure conditions. A slight, but significant CrPic-induced increase in DNA damage (P<0.001) was observed in the human lymphocytes, but only when these cells were exposed in the absence of serum. In all other experiments CrPic was found to be without genotoxic effects, both in vivo and in vitro. Taken together, our results suggest that a high concentration of CrPic might be DNA damaging, but only under non-physiological conditions. PMID- 17418472 TI - Behavioral and immunohistological assessment of painful neuropathy induced by a single oxaliplatin injection in the rat. AB - In clinical use, a single infusion of oxaliplatin, widely used to treat metastatic colorectal cancer, induces specific sensory neurotoxicity signs triggered or aggravated by exposure to cold. To study the pathophysiology of these symptoms, we developed and characterized an animal model that reproduces the effects of a single intraperitoneal oxaliplatin administration (3, 6 and 12 mg/kg). Significant allodynia and hyperalgesia to cold stimuli were rapidly observed from 24 h to day 5 with a maximum lowering of 76% at t+30 h versus control. Other behavioral assessments revealed rapid persistent mechanical allodynia, but no thermal hyperalgesia or allodynia to heat and no hyperalgesia to mechanical stimuli. An immunohistochemical study in the superficial layers of the spinal dorsal horn revealed a marked increase in substance P immunoreactivity versus controls (12% versus 4%), whereas calcitonin gene-related peptide (CGRP) immunoreactivity was unchanged. This new animal model for the first time closely mimics the effects observed in humans after a single oxaliplatin infusion, especially onset and highly intense sensory disturbances, hypersensitivity to cold with allodynia and hyperalgesia signs. This model may help to elucidate the mechanisms of this thermal hypersensitivity, especially the possible involvement of small-diameter A-fibers in cold allodynia symptoms. These selective effects may clue up the mechanistic basis for the acute oxaliplatin neuropathy leading to a better understanding of the clinical condition and to optimize its treatment. PMID- 17418473 TI - Low concentrations of ketamine initiate dendritic atrophy of differentiated GABAergic neurons in culture. AB - Administration of subanesthetic concentrations of ketamine, a noncompetitive antagonist of the N-methyl-d-aspartate (NMDA) type of glutamate receptors, is a widely accepted therapeutic modality in perioperative and chronic pain management. Although extensive clinical use has demonstrated its safety, recent human histopathological observations as well as laboratory data suggest that ketamine can exert adverse effects on central nervous system neurons. To further investigate this issue, the present study was designed to evaluate the effects of ketamine on the survival and dendritic arbor architecture of differentiated gamma aminobutyric acidergic (GABAergic) interneurons in vitro. We show that short-term exposure of cultures to ketamine at concentrations of > or =20 microg/ml leads to a significant cell loss of differentiated cells and that non-cell death-inducing concentrations of ketamine (10 microg/ml) can still initiate long-term alterations of dendritic arbor in differentiated neurons, including dendritic retraction and branching point elimination. Most importantly, we also demonstrate that chronic (>24 h) administration of ketamine at concentrations as low as 0.01 microg/ml can interfere with the maintenance of dendritic arbor architecture. These results raise the possibility that chronic exposure to low, subanesthetic concentrations of ketamine, while not affecting cell survival, could still impair neuronal morphology and thus might lead to dysfunctions of neural networks. PMID- 17418474 TI - Accuracy of contacts calculated from 3D images of occlusal surfaces. AB - OBJECTIVES: Compare occlusal contacts calculated from 3D virtual models created from clinical records to contacts identified clinically using shimstock and transillumination. METHODS: Upper and lower full arch alginate impressions and vinyl polysiloxane centric interocclusal records were made of 12 subjects. Stone casts made from the alginate impressions and the interocclusal records were optically scanned. Three-dimensional virtual models of the dental arches and interocclusal records were constructed using the Virtual Dental Patient Software. Contacts calculated from the virtual interocclusal records and from the aligned upper and lower virtual arch models were compared to those identified clinically using 0.01mm shimstock and transillumination of the interocclusal record. Virtual contacts and transillumination contacts were compared by anatomical region and by contacting tooth pairs to shimstock contacts. Because there is no accepted standard for identifying occlusal contacts, methods were compared in pairs with one labeled "standard" and the second labeled "test". Accuracy was defined as the number of contacts and non-contacts of the "test" that were in agreement with the "standard" divided by the total number of contacts and non-contacts of the "standard". RESULTS: Accuracy of occlusal contacts calculated from virtual interocclusal records and aligned virtual casts compared to transillumination were: 0.87+/-0.05 and 0.84+/-0.06 by region and 0.95+/-0.07 and 0.95+/-0.05 by tooth, respectively. Comparisons with shimstock were: 0.85+/-0.15 (record), 0.84+/-0.14 (casts), and 81+/-17 (transillumination). CONCLUSIONS: The virtual record, aligned virtual arches, and transillumination methods of identifying contacts are equivalent, and show better agreement with each other than with the shimstock method. PMID- 17418475 TI - Serial position effects in free memory recall--An ERP-study. AB - Event-related brain potentials (ERPs) elicited by recalled and non-recalled words were recorded from 18 female subjects to investigate primacy and recency effects in free memory recall. The typical pattern of a serial position curve (SPC) was obtained with words presented at first and final positions in a list recalled better than words presented in the middle of a list. A marked positivity is seen in the ERPs for words on Primacy, but not on Recency positions at frontocentral electrodes. In contrast, ERP amplitudes on parietal electrodes resemble the SPC seen in behavioral data: P300 amplitude is largest for words on Primacy and Recency positions and attenuated on Plateau positions. Furthermore, subjects with a clear Primacy effect in behavioral data show a distinct frontal positive slow wave for Primacy words only, whereas subjects without a clear primacy effect show a frontal "difference due to subsequent memory" (DM) effect for Primacy and Plateau words. These results are discussed in the framework of working memory and distinctiveness. PMID- 17418477 TI - Influence of N-H...O and O-H...O hydrogen bonds on the (17)O, (15)N and (13)C chemical shielding tensors in crystalline acetaminophen: a density functional theory study. AB - A computational investigation was carried out to characterize the (17)O, (15)N and (13)C chemical shielding tensors in crystalline acetaminophen. We found that N-H...O and O-H...O hydrogen bonds around the acetaminophen molecule in the crystal lattice have different influences on the calculated (17)O, (15)N and (13)C chemical shielding eigenvalues and their orientations in the molecular frame of axes. The calculations were performed with the B3LYP method and 6 311++G(d, p) and 6-311+G(d) standard basis sets using the Gaussian 98 suite of programs. Calculated chemical shielding tensors were used to evaluate the (17)O, (15)N, and (13)C NMR chemical shift tensors in crystalline acetaminophen, which are in reasonable agreement with available experimental data. The difference between the calculated NMR parameters of the monomer and molecular clusters shows how much hydrogen-bonding interactions affect the chemical shielding tensors of each nucleus. The computed (17)O chemical shielding tensor on O(1), which is involved in two intermolecular hydrogen bonds, shows remarkable sensitivity toward the choice of the cluster model, whereas the (17)O chemical shielding tensor on O(2) involved in one N-H...O hydrogen bond, shows smaller improvement toward the hydrogen-bonding interactions. Also, a reasonably good agreement between the experimentally obtained solid-state (15)N and (13)C NMR chemical shifts and B3LYP/6-311++G(d, p) calculations is achievable only in molecular cluster model where a complete hydrogen-bonding network is considered. Moreover, at the B3LYP/6-311++G(d, p) level of theory, the calculated (17)O, (15)N and (13)C chemical shielding tensor orientations are able to reproduce the experimental values to a reasonably good degree of accuracy. PMID- 17418478 TI - A quantitative framework for the design of acellular hemoglobins as blood substitutes: implications of dynamic flow conditions. AB - The delivery of oxygen to tissue by cell-free carriers eliminates intraluminal barriers associated with red blood cells. This is important in arterioles, since arteriolar tone controls capillary perfusion. We describe a mathematical model for O(2) transport by hemoglobin solutions and red blood cells flowing through arteriolar-sized tubes to optimize values of p50, Hill number, hemoglobin molecular diffusivity and concentration. Oxygen release is evaluated by including an extra-luminal resistance term to reflect tissue oxygen consumption. For low consumption (i.e., high resistance to O(2) release) a hemoglobin solution with p50=15 mmHg, n=1, D(HBO2)=3 x 10(-7) cm(2)/s delivers O(2) at a rate similar to that of red blood cells. For high consumption, the p50 must be decreased to 5 mmHg. The model predicts that regardless of size, hemoglobin solutions with higher p50 will present excess O(2) to arteriolar walls. Oversupply of O(2) to arteriolar walls may cause constriction and paradoxically reduced capillary perfusion. PMID- 17418479 TI - Ions in water: characterizing the forces that control chemical processes and biological structure. AB - The continuum electrostatics model of Debye and Huckel [P. Debye and E. Huckel, On the theory of electrolytes. I. Freezing point depression and related phenomena., Phys. Z. 24 (1923) 185-206.] and its successors utilize a macroscopic dielectric constant and assume that all interactions involving ions are strictly electrostatic, implying that simple ions in water generate electric fields strong enough to orient water dipoles over long distances. However, solution neutron and X-ray diffraction indicate that even di- and tri-valent ions do not significantly alter the density or orientation of water more than two water molecules (5 A) away. Therefore the long range electric fields (generated by simple ions) which can be detected by various resonance techniques such as fluorescence resonance energy transfer over distances of 30 A (about 11 water diameters) or more must be weak relative to the strength of water-water interactions. Two different techniques indicate that the interaction of water with anions is by an approximately linear hydrogen bond, suggesting that the dominant forces on ions in water are short range forces of a chemical nature. PMID- 17418480 TI - Development of a bioremediation process by biostimulation of native microbial consortium through the heap leaching technique. AB - Heap leaching is an effective and widely used method of recovering metals from low-grade ores. However, the heap leaching technique has not yet been used in other biotechnological processes such as bioremediation. This work describes biostimulation of the native microbial consortium as a novel application of the heap leaching technique to bioremediate mining soils contaminated with hydrocarbons. Microorganisms present in the polluted soil were isolated in a liquid mineral solution using diesel fuel as the sole energy and carbon source. Biodegradation activity was evaluated and two genera, Flavobacterium and Aspergillus, were identified as the primary microorganisms that degraded hydrocarbons in the polluted soil. In order to simulate the heap leaching process on a laboratory scale, using both columns and piles, the contaminated soil was mixed with different sand concentrations and was agglomerated before it was used. Three flow rates, of the mineral solution, were evaluated. Of the rates tested, biodegradation was most efficient at a flow rate of 200 ml h(-1). The heap leaching technique demonstrated good efficiency in the column and pile, with a 2% soil-sand mixture lowering the TPH concentration from 61,000 to 1800 mg kg(-1) (98.5%) in 15 d. PMID- 17418481 TI - Two-stage transperineal management of posterior urethral strictures or bladder neck contractures associated with urinary incontinence after prostate surgery and endoscopic treatment failures. AB - OBJECTIVES: The treatment of posterior urethral strictures or bladder neck contracture associated with severe urinary incontinence after prostate surgery and failure of endoscopic treatments is controversial. We report our experience with a transperineal approach in two steps: end-to-end urethroplasty/anastomosis and subsequent artificial urinary sphincter implantation. METHODS: Between September 2001 and January 2005, we observed six patients (58-68 yr old), with a combination of severe urinary incontinence and posterior urethral stricture with anastomotic bladder neck contracture after prostate surgery. In all cases, repeated endoscopic treatments of the strictures failed. The patients underwent transperineal end-to-end urethroplasty or anastomosis followed by transperineal artificial urinary sphincter placement after 6 mo. RESULTS: After the first surgical step, all patients were completely incontinent with absence of urethral strictures and complete anastomotic healing in all cases. Therefore, all patients underwent artificial urinary sphincter insertion. After a mean follow-up of 38 mo (range: 18-57 mo), five patients are continent with no postvoid residual urine and a perfectly functioning device. One artificial urinary sphincter was removed due to urethral erosion. CONCLUSIONS: In patients with posterior urethral strictures or bladder neck contractures associated with severe urinary incontinence, an artificial urinary sphincter implantation as a second step allows verification of the outcome of a previous end-to-end urethroplasty or anastomosis and utilizes a dedicated operative field to reduce the risks of prosthesis implants. PMID- 17418482 TI - Clinical relevance of transurethral resection of the prostate in "asymptomatic" patients with an elevated prostate-specific antigen level. AB - OBJECTIVES: To determine the clinical relevance of transurethral resection of the prostate (TURP) in patients with minor lower urinary tract symptoms (LUTS) but elevated prostate-specific antigen (PSA) levels. METHODS: We retrospectively included 82 patients, aged 50.2-78.2 yr, with minor LUTS, elevated PSA (> or =4 ng/ml), and no signs of prostate cancer (PCa) after (multiple) negative multisite biopsies who underwent TURP after they were diagnosed by urodynamics with bladder outlet obstruction (BOO). We evaluated the clinical benefit of TURP by assessing its effect on International Prostate Symptom Score (IPSS) and PSA and the diagnostic value of histologic examination of the resected tissue for the presence of PCa. RESULTS: After TURP, histologic analysis of the resected specimen revealed that eight patients (9.8%) had PCa; seven of these patients had a tumour that needed further treatment. The remaining 74 patients (90.2%) were diagnosed with BOO due to benign prostatic hyperplasia/benign prostatic enlargement (BPH/BPE). In this group, the mean PSA level decreased from 8.8 ng/ml before TURP to 1.1 ng/ml in the first year and 1.3 ng/ml in the second year after TURP; the mean IPSS decreased from 8.8 to 1.5 in the first year after TURP. CONCLUSIONS: The current data suggest that patients with minor LUTS and elevated PSA without evidence of PCa are very likely to have BOO due to BPH/BPE and may benefit from TURP if obstruction is proved. However, a prospective trial is warranted to assess the impact of these results on clinical practice. PMID- 17418483 TI - A study on emergence of alternating reciprocity in a 2 x 2 game with 2-length memory strategy. AB - It is recognized that bilateral cooperation (C), a reward-state in other words, emergently comes up in a 2 x 2 prisoner's dilemma game, if you assume a strategy set with a memory concept. Also observed is a mixture state of cooperation (C) and defect (D), saint- and temptation-state in other words, to obtain a higher payoff than R (R reciprocity) in a hero or leader game that is a chicken-type dilemma game; this phenomenon is called alternating reciprocity (AR) or ST reciprocity. Observing a holistic 2 x 2 game world including trivial games and various dilemma games, where 2-length memory and infinite interactions are assumed, the paper reports on the specific mechanism of AR. It is observed there are three different phases relating to AR, which can be explained by the stress of the dilemma. PMID- 17418484 TI - Carotid plaque inflammation detected by 18F-fluorodeoxyglucose-positron emission tomography. Pilot study. AB - OBJECTIVES: Inflammation is important in both the pathogenesis and outcome of atherosclerosis. Current imaging techniques provide anatomic data but no indication of plaque inflammation. We tested the hypothesis that plaque inflammation could be assessed in vivo by (18)FDG-PET and that plaque inflammation could increase the risk of recurrent vascular events and poor response to treatment in a pilot study. PATIENTS AND METHODS: Thirteen patients (median age 66.1 years [55-82 years]) with recent carotid territory TIA or ischemic stroke and internal carotid artery (ICA) stenosis > or =50% were studied. Angiography and (18)F-fluorodeoxyglucose-positron emission tomography ((18)FDG-PET) imaging were carried out in all patients. Treatment for carotid stenosis in each patient was selected by the attending physician and consisted in medical treatment, endarterectomy or stent placement. During 6 months of follow up, the specific end points assessed were the occurrence of any stroke, death, or re-stenosis. RESULTS: Patients with symptomatic carotid atherosclerosis were imaged using (18)FDG-PET. Strong (18)FDG uptake (SUV> or =2.7) was seen in 11 of 13 (85%) carotid lesions. Among these patients two died during follow-up, 3 had recurrent non-fatal ipsilateral ischemic stroke and 1 patient who had undergone stenting had non-symptomatic re-stenosis in control studies. There was a significant correlation between the (18)FDG uptake and degree of ICA stenosis detected by angiography. CONCLUSION: Carotid atherosclerotic plaques contain a variable degree of inflammation which can be assessed in vivo by means of FDG and PET. The prognostic value of this marker is, however, still unclear and needs further study. PMID- 17418485 TI - Wheat phytotoxicity from arsenic and cadmium separately and together in solution culture and in a calcareous soil. AB - The toxicity of two toxic elements, arsenic (As) and cadmium (Cd) (individually or in combination) on root elongation of wheat seedlings (Triticum aestivum, L.) were investigated both in hydroponics and in soils freshly spiked with the toxic elements. Median effective concentration (EC(50)) and non-observed effect concentration (NOEC) were used to investigate the toxic thresholds and potencies of the two elements. The EC(50) for As was 0.97 microM in hydroponics and 196 mgkg(-1) in soil, and 4.32 microM and 449 mgkg(-1) for Cd, respectively. Toxic unit (TU) and additive index (AI) concepts were introduced to determine the combined outcomes, and different behaviors were obtained: synergism in solution culture (EC(50 mix)=0.36 TU(mix) and AI: 1.76) and antagonism in soil experiments (EC(50 mix)=1.49 TU(mix) and AI: -0.33). Furthermore, the data of soil bioavailable As and Cd cannot explain the discrepancy between the results derived from soil and hydroponics experiments. PMID- 17418486 TI - Speciation and determination of thallium by on-line microcolumn separation/preconcentration by flow injection-flame atomic absorption spectrometry using immobilized oxine as sorbent. AB - A flow injection analysis (FIA) system incorporation a microcolumn of immobilized oxine on surfactant-coated alumina had been devised for performing rapid thallium enrichment/matrix removal in flame atomic absorption spectrometry (FAA). The preconcentration is based on the deposition of thallium(I) on microcolumn and subsequent elution with 500 microl of sodium thiosulfate (1 moll(-1)). In the presence of EDTA, only Tl(I) was retained on the microcolumn. Total thallium was determined after reduction of Tl(III) to Tl(I) by hydroxyl amine hydrochloride. A sample volume of 25 ml resulted in a preconcentration factor of 77. Precision at 30 microgl(-1) was 2.6% RSD (n=10). With 25 ml sampling volume a detection limit of 2.5 microgl(-1) was determined. The effect of potential interfering ions on the determination was studied. The method was applied for the determination of thallium in water, waste water, hair, nail, coal, and standard reference alloys. The accuracy was assessed through recovery experiment, independent analysis by Furnace-AAS, and analysis of certified reference alloys. PMID- 17418487 TI - The use of 2D non-uniform electric field to enhance in situ bioremediation of 2,4 dichlorophenol-contaminated soil. AB - In situ bioremediation is a safe and cost-effective technology for the cleanup of organic-contaminated soil, but its remediation rate is usually very slow, which results primarily from limited mass transfer of pollutants to the degrading bacteria in soil media. This study investigated the feasibility of adopting 2D non-uniform electric field to enhance in situ bioremediation process by promoting the mass transfer of organics to degrading bacteria under in situ conditions. For this purpose, a 2D non-uniform electrokinetic system was designed and tested at bench-scale with a sandy loam as the model soil and 2,4-dichlorophenol (2,4-DCP) as the model organic pollutant at two common operation modes (bidirectional and rotational). Periodically, the electric field reverses its direction at bidirectional mode and revolves a given angle at rotational mode. The results demonstrated that the non-uniform electric field could effectively stimulate the desorption and the movement of 2,4-DCP in the soil. The 2,4-DCP was mobilized through soil media towards the anode at a rate of about 1.0 cmd(-1)V(-1). The results also showed that in situ biodegradation of 2,4-DCP in the soil was greatly enhanced by the applied 2D electric field upon operational mode. At the bidirectional mode, an average 2,4-DCP removal of 73.4% was achieved in 15 days, and the in situ biodegradation of 2,4-DCP was increased by about three times as compared with that uncoupled with electric field, whereas, 34.8% of 2,4-DCP was removed on average in the same time period at the rotational mode. In terms of maintaining remediation uniformity in soil, the rotational operation remarkably excelled the bidirectional operation. In the hexagonal treatment area, the 2,4 DCP removal efficiency adversely increase with the distance to the central electrode at the bidirectional mode, while the rotational mode generated almost uniform removal in soil bed. PMID- 17418488 TI - Preparation, characterization of cellulose-grafted with calix[4]arene polymers for the adsorption of heavy metals and dichromate anions. AB - In this paper, the adsorbents were prepared from cellulose-grafted with calix[4]arene polymers (CGC[4]P-1 and CGC[4]P-2) and their sorption properties studied. The polymers were characterized by Fourier transform infrared spectroscopy, elemental analysis, thermal gravimetric analysis and scanning electron microscopy. They were then used to evaluate the sorption properties of some heavy metal cations (Co(2+), Ni(2+), Cu(2+), Cd(2+), Hg(2+) and Pb(2+)) and dichromate anions (Cr(2)O(7)(2-)/HCr(2)O(7)(-)). Results showed that CGC[4]P-2 was a good sorbent for heavy metal cations while CGC[4]P-1 was ineffective. In the studies of dichromate anion sorption, it was observed that CGC[4]P-2 was a more highly effective sorbent at pH 1.5 than was CGC[4]P-1. PMID- 17418489 TI - Response inhibition in psychopathy: the frontal N2 and P3. AB - Psychopathy has been associated with atypical function of the anterior cingulate cortex (ACC) and adjacent brain regions and with abnormalities in performance monitoring, which is thought to rely on these structures. The ACC and adjacent regions are also involved in the generation of two characteristic components of the event-related potential: the frontal N2 and P3. Both components are enhanced when a response is withheld (NoGo trial) within a series of positive-responses (Go trials) and are considered an index of response inhibition. We recorded event related potentials while violent offenders who varied on the dimension of psychopathy and non-offender controls performed a Go/NoGo task. The offenders made more errors of commission on NoGo trials but this effect was unrelated to level of psychopathy within the group and, inconsistent with a previous report, they produced the enhanced frontal N2 and P3 effect in response to NoGo relative to Go conditions. We conclude that the neural processes involved in response inhibition are not abnormal in psychopaths when both stimuli and context are affectively neutral and suggest that a more nuanced perspective regarding impulsivity in this population be considered. PMID- 17418490 TI - Reduction of EphA4 receptor expression after spinal cord injury does not induce axonal regeneration or return of tcMMEP response. AB - Spinal cord injury (SCI) causes an increase of inhibitory factors that may restrict axonal outgrowth after trauma. During the past decade, the Eph receptors and ephrin ligands have emerged as key repulsive cues known to be involved in neurite outgrowth, synapse formation, and axonal pathfinding during development. Given the non-permissive environment for axonal regeneration after SCI, we questioned whether enhanced-expression of the EphA4 receptor with repulsive activity for axonal outgrowth is potentially responsible for the regenerative failure. To address this possibility, we have examined the expression of EphA4 after SCI in adult rats following a contusion SCI. EphA4 expression studies demonstrated a time-dependent change for EphA4 protein without alterations in beta-actin. EphA4 was downregulated initially and upregulated 7 days after injury. Blockade of EphA4 upregulation with antisense oligonucleotides did not produce an anatomical or physiological response monitored with anterograde tracing studies or transcranial magnetic motor evoked potentials (tcMMEP), respectively. These results demonstrated that upregulation of EphA4 receptors after trauma is not related to axonal regeneration or return of nerve conduction across the injury site. PMID- 17418491 TI - Poor inheritance of low attractiveness for Amblyomma variegatum in cattle. AB - Because of the high variability of cattle infestation, selective breeding has been proposed to control three-host ticks in Africa in the same way as it has been implemented to control the one-host tick Boophilus microplus in Australia. Interbreeding (divergent selection experiment) between Gudali zebu cattle was performed at the Wakwa Research Centre, Adamawa, Cameroon to assess the feasibility and impact of such a selective breeding programme. Fifty cows (chosen out of 129) and 2 bulls (chosen out of 10) were selected according to their greater or lesser attractiveness for the tick Amblyomma variegatum, assessed by their infestation degree (ID, ratio between individual animal infestation and mean herd infestation). Half of the animals displayed a high ID (H) and the remainder a low ID (L). The presence of a suckling calf had a significant effect on the infestation of its dam, the udder of lactating cows being significantly less infested (P<0.001) because of a tick removal effect by the calves. Observed infestation data were therefore corrected to allow comparison between lactating and non-lactating cows. The ID of the cows was assessed on five occasions from 1992 to 1994: the consistency of the infestation hierarchy was good overall (P<0.001) despite high variability seen in some animals which would complicate the selection of cattle with low infestation. Four breeding groups were set up (LxL, LxH, HxL, HxH) and 40 calves were born in 1993 and 1994. There was a correlation (P<0.02-0.001) between liveweight (or age) and ID when the younger calves of the groups were 6 months old, but the correlation disappeared by the time the calves were all older than 1 year. There was no difference between the average ID of the calves from the four breeding groups, and no correlation was observed between dam ID and calf ID. The average ID of the 10 calves born of the dams with the lowest ID was however significantly lower than that of the 10 calves born of the dams with the highest ID (0.82 versus 1.15, P=0.03), but the difference was much less than that observed between the dams (0.35 versus 2.30, P=7x10(-9)). A positive correlation between dam ID and calf ID was found when only these 20 cows were taken into consideration (r=0.57, P<0.01). These results indicate that selective breeding of Gudali zebu cattle to produce animals with low infestation by the tick A. variegatum would be uncertain but that a more drastic selection of the breeders than that implemented in the present study may perhaps have some positive impact. PMID- 17418492 TI - Viability and infectivity of Trichinella spiralis muscle larvae in frozen horse tissue. AB - Many aspects of the biology and epidemiology of Trichinella infection in the horse are poorly understood, including survival of Trichinella spp in horse muscle. In this study, we have assessed the freeze tolerance of T. spiralis in horse meat stored at 5, -5, and -18 degrees C for 1 day to 24 weeks. Results demonstrate a steady reduction in the number of live ML recovered from the cold stored meat samples. On Day 1, recovery of live larvae had been reduced by 18.6%, 50.1%, and 37.2%, and by 4 weeks, recovery of larvae had been reduced by 65.4%, 66.5%, and 96.2% in samples stored at 5, -5, and -18 degrees C, respectively. Infectivity results (measured as reproductive capacity index (RCI)) from mice inoculated with larvae recovered from non-frozen meat samples at day 0 was 23.5. Following storage at -18 degrees C for one and two days, the RCIs were 2.09 and 0.99, respectively. Small numbers of infective larvae were still present in meat samples stored at -18 degrees C for 4 weeks. The RCI of ML recovered from meat samples stored at -5 degrees C was 14.99 and 6.36 at 2 weeks and 4 weeks respectively; the RCI of samples stored at 5 degrees C was 23.1 at 8 weeks, and fell rapidly thereafter (12 week RCI 1.33; 0 at 24 weeks). These data demonstrate that infective T. spiralis, a non-freeze tolerant species, can survive for at least 4 weeks in horse tissue frozen at -5 or -18 degrees C, and that the numbers of infective larvae decrease substantially by day 2 at -18 degrees C and by week 4 at -5 degrees C. PMID- 17418493 TI - Adverse effects of over the counter cryotherapy. PMID- 17418494 TI - P19 embryonal carcinoma cells as in vitro model for studying purinergic receptor expression and modulation of N-methyl-D-aspartate-glutamate and acetylcholine receptors during neuronal differentiation. AB - The in vitro differentiation of P19 murine embryonal carcinoma cells to neurons resembles developmental stages which are encountered during neuronal development. Three days following induction to neuronal differentiation by retinoic acid, most cells of the P19 population lost expression of the stage specific embryonic antigen (SSEA-1) and expressed the neural progenitor cell specific antigen nestin. Beginning from day 4 of differentiation nestin expression was down regulated, and expression of neuron-specific enolase as marker of differentiated neurons increased. The molecular mechanisms underlying neuronal differentiation are poorly understood. We have characterized the participation of purinergic ionotropic (P2X) and metabotropic (P2Y) receptors at mRNA transcription and protein levels as well as ATP-induced Ca2+ transients during neuronal differentiation of P19 cells. Gene and protein expression of P2X2, P2X6, P2Y2, and P2Y6 receptors increased during the course of differentiation, whereas P2X3, P2X4, P2Y1 and P2Y4 receptor expression was high in embryonic P19 cells and then decreased following induction of P19 cells to differentiation. P2X1 receptor protein expression was only detected on days 2 and 4 of differentiation. Although P2X5 and P2X7 mRNA transcription was present, no protein expression for this receptor subunit could be detected throughout the differentiation process. In undifferentiated cells, mainly ionotropic P2X receptors contributed to the ATP induced Ca2+-response. In neuronal-differentiated P19 cells, the ATP-induced Ca2+ response was increased and the metabotropic component predominated. Purinergic receptor function is implicated to participate in neuronal maturation, as cholinergic and glutamate-N-methyl-D-aspartate (NMDA) induced calcium responses were affected when cells were differentiated in the presence of purinergic receptor antagonists pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS), suramin or reactive blue-2. Our data suggest that inhibition of P2Y1 and possibly P2X2 receptors led to a loss of NMDA receptor activity whereas blockade of possibly P2X2 and P2Y2 purinergic receptors during neuronal differentiation of P19 mouse led to inhibition of cholinergic receptor responses. PMID- 17418495 TI - Respiratory activity in brainstem of fetal mice lacking glutamate decarboxylase 65/67 and vesicular GABA transporter. AB - The respiratory neural network in the mammalian medulla oblongata shows rhythmic activity before birth. GABA and glycine are considered to be involved in control of respiratory rhythm. Recently we have demonstrated respiratory failure in glutamic acid decarboxylase (GAD) 67-deficient mice [Tsunekawa N, Arata A, Obata K (2005) Development of spontaneous mouth/tongue movement and related neural activity, and their repression in mouse fetus lacking glutamate decarboxylase 67. Eur J Neurosci 21:173-178]. To further evaluate the involvement of GABA and glycine in fetal respiratory function, we studied neural activities in brainstem spinal cord blocks prepared from GAD65-/-:67-/- and vesicular GABA transporter (VGAT)-/-mice on embryonic day 14 (E14)-E15 and E18. In these knockout mice, the synthesis of GABA and the vesicular release of GABA and glycine are completely absent, respectively. Spontaneous respiratory discharges were observed in the ventral roots at the cervical cord (C) 4 level from wild-type mice but not from the knockout mice on E18. Administration of substance P induced C4 discharges in GAD65-/-:67-/- preparations but not in VGAT-/- preparations. C4 discharges were observed in the knockout mice on E14-E15, although the frequency was lower than that in the wild-type. Neuronal activities in the respiratory network of the E18 brainstem were recorded using a "blind" patch-clamp technique. Expiratory and inspiratory neurons with their characteristic firing patterns were observed in the wild-type fetuses. Strychnine reversed inspiratory-phase hyperpolarization to large depolarization in expiratory neurons. On the other hand, neurons in the same area of the knockout mice fired spontaneously without any rhythm. Substance P induced hyperpolarizing potentials in medullary neurons of GAD65-/-:67-/- mice. Further administration of strychnine induced large depolarizing potentials. Rhythmic activities were not observed in VGAT-/- mice even in the presence of substance P and strychnine. These results indicate that the lack of GABA and glycine impairs the function of the respiratory network in mouse fetuses and the impairment progresses with fetal age. PMID- 17418496 TI - Feedback modulates the temporal scale-free dynamics of brain electrical activity in a hypothesis testing task. AB - We used the electroencephalogram (EEG) to investigate whether positive and negative performance feedbacks exert different long-lasting modulations of electrical activity in a reasoning task. Nine college students serially tested hypotheses concerning a hidden rule by judging its presence or absence in triplets of digits, and revised them on the basis of an exogenous performance feedback. The scaling properties of the transition period between feedback and triplet presentation were investigated with detrended fluctuation analysis (DFA). DFA showed temporal scale-free dynamics of EEG activity in both feedback conditions for time scales larger than 150 ms. Furthermore, DFA revealed that negative feedback elicits significantly higher scaling exponents than positive feedback. This effect covers a wide network comprising parieto-occipital and left frontal regions. We thus showed that specific task demands can modify the temporal scale-free dynamics of the ongoing brain activity. Putative neural correlates of these long-lasting feedback-specific modulations are proposed. PMID- 17418497 TI - Anti-nociceptive effects of selectively destroying substance P receptor expressing dorsal horn neurons using [Sar9,Met(O2)11]-substance P-saporin: behavioral and anatomical analyses. AB - Lumbar intrathecal injections of substance P-saporin (SP-sap) destroy dorsal horn neurons that express the neurokinin-1 receptor (NK-1R) resulting in decreased responses to a range of noxious stimuli and decreased hyperalgesia and allodynia. Forebrain injections of SP-sap produce considerable non-specific damage raising some concern about use of this toxin in vivo. The more stable and selective substance P congener, [Sar9,Met(O2)11]substance P coupled to saporin (SSP-sap) produces much more selective forebrain lesions at significantly lower doses. The present study sought to determine the anatomic and nocifensive behavioral effects of lumbar intrathecal injections of the more precisely targeted SSP-sap. On the basis of loss of lamina I NK-1R staining, lumbar intrathecal SSP-sap was seven times more potent than SP-sap and produced no loss of NK-1R expressing neurons in deeper laminae (III-VI or X). Transient decreases in hotplate responding occurred at 44 degrees C and 47 degrees C but not 52 degrees C during the first 3 weeks after SSP-sap injection with return to baseline by 4 weeks. Operant escape responses were reduced at 0.3 degrees C, 44 degrees C and 47 degrees C for at least 4 months. In the formalin test, SSP-sap also was about seven times more potent than SP-sap in reducing phase two behavior in both female Long Evans and male Sprague-Dawley rats. Both SSP-sap and SP-sap reduced formalin-induced FOS expression in deep and superficial laminae of the L4 dorsal horn in parallel with the reduction in phase 2 behavior. In summary, SSP-sap is highly effective in destroying lamina I NK-1R expressing neurons, without loss of deep NK-1R neurons. The behavioral effects of SSP-sap are similar to SP-sap suggesting that the antinociceptive effects of both toxins are indeed due to selective loss of NK-1R neurons in lamina I. SSP-sap is an attractive agent for possible treatment of chronic pain. PMID- 17418498 TI - Determinants of the diurnal course of salivary alpha-amylase. AB - OBJECTIVE: Previous data from our group and others have shown that salivary alpha amylase activity increases in response to stress. It has been suggested that salivary alpha-amylase may be a marker for adrenergic activity. Less is known about other determinants of salivary alpha-amylase activation. The objective of the current study was to describe the diurnal pattern of salivary amylase and its determinants. METHODS: Saliva samples were collected immediately after waking-up, 30 and 60 min later, and each full hour between 0900 and 2000 h by 76 healthy volunteers (44 women, 32 men). Compliance was controlled by electronic monitors. In order to control factors which might influence the diurnal profile of salivary alpha-amylase (such as momentary stress, mood, food, or body activity), at each sampling time point the subjects filled out a diary examining the activities they had carried out during the previous hour. RESULTS: Salivary alpha-amylase activity shows a distinct diurnal profile pattern with a pronounced decrease within 60 min after awakening and a steady increase of activity during the course of the day. Mixed models showed a relative independence of diurnal salivary alpha amylase from momentary stress and other factors, but significant associations with chronic stress and mood. CONCLUSIONS: Our results suggest that diurnal profiles of salivary alpha-amylase are relatively robust against momentary influences and therefore may prove useful in the assessment of sympathetic nervous system activity. The findings underscore the need to control for time of day in studies using salivary alpha-amylase as a dependent variable. PMID- 17418499 TI - Pushing 'bad bugs' into committing suicide: activation of microbial autolysis within the gut using food-grade substances as a prospective method for treatment of intestinal infections and/or immunity enhancement. PMID- 17418500 TI - Application of Pegaptanib on spinal cord lesions and central nervous system lesions to induce neuronal regeneration. PMID- 17418501 TI - Follicular dendritic cell hyperplasia in plasma cell variant of Castleman's disease with interfollicular Hodgkin's disease. AB - We report a case of a multicentric plasma cell (PC) variant of Castleman's disease (CD) in association with interfollicular type of classic Hodgkin's disease (HD), both diseases identified in the same lymph node. The histologic features of CD were the classic ones, with hyperplastic and atrophic follicles, some with prominent mantle zones, hyalinzed vessels, and a very rich polyclonal proliferation of PCs in the interfollicular region. The presence of LCA-negative, but CD30- and CD15-positive typical and atypical Reed-Sternberg (RS) cells in the interfollicular region confirmed the presence of HD. In addition, many of the RS cells stained positive for EBV. CD35- and CD21-positive follicular dendritic cell (FDC) hyperplasia was a striking feature, a finding that has not been well documented in the PC variant of CD. PMID- 17418502 TI - [Different human cytomegalovirus glycoprotein B (gB) genotype distribution]. AB - The glycoprotein B (gB) is the major glycoprotein of the envelope of the human cytomegalovirus, it is encoded by UL55 open reading frame, implicated in host cell, entry cell to cell virus transmission and fusion of infected cells and a significant is an important target for immuno-reactions humoral and cellular. A prospective analysis of cytomegalovirus glycoprotein B genotype was conducted on 31 immunodepressed (transplant recipients and AIDS patients). The DNA was extracted directly from the bronchoalveolar liquid (BAL) of these patients. The gB genotypes of CMV was determined by using the polymerase chain reaction, followed by the digestion of two enzymes of restriction HinfI and RsaI. The distribution of the gB genotype of the CMV was: gB1 38,70%; gB2 25,80%; gB3 16,12% and gB4 19,35%. The analysis of the peptide sequence of this region (codons 437-520), indicate the variation was more frequent between codons 448 480. PMID- 17418503 TI - [Genotypes of echovirus 6 in Monastir region]. AB - OBJECTIVE OF THE WORK: Echoviruses of serotype 6 were reported to be endemic in Tunisia and even in other country over the world. they are associated with many outbreak meningitis. The Objective of this study was to genetically characterize echovirus 6 fields isolates. It gives a first approach on the molecular epidemiology of this serotype. MATERIAL AND METHODS: Phylogenetic analysis of partial sequence in the 3'half of the VP1 region (2874-3529) from 25 strains of echovirus 6. RESULTS: 9 genotypes of echovirus 6 were individualized. Study area was Monastir, a touristic tunisian city. Strains were isolated from wastewater during one year, may correspond to three genotypes. CONCLUSION: Many genotype could circulating during the same time and in the same region. This phenomena was reported to be atypic in the case of poliovirus. PMID- 17418504 TI - [Interest of chronotherapy in multidisciplinary management of oesophageal and gastric cancers]. AB - The authors evaluated the impact of a chronotherapy with 5-FU, folinic acid and carboplatine (chronomodulated infusions by ambulatory pumps; 5/21 days) for the management of oesophagus (52 cases) and gastric (56 cases) cancer patients. The overall tolerance of treatment was gauged excellent (grade 3-4; % patients: mucitis: 11-23%; leucopenia 6-19%; thrombopenia 18-50%; almost no digestive disturbances nor alopecia). Also tumor outcome was considered interesting with major responses rate in 61% (gastric) to 79% (oesophagus) of patients. The median survival of oesophageal cancer was limited to 9.2 months; the one of disseminated gastric cancer was 12.7 months but 72% of curatively resected patients were alive at 5+ years. PMID- 17418505 TI - An advantage for concavities in shape perception by chimpanzees (Pan troglodytes). AB - The significance of concavity in object shape perception by chimpanzees (Pan troglodytes) was investigated in a matching-to-sample procedure. For the task, chimpanzees were required to choose a polygon stimulus that was identical in shape to a sample. The incorrect alternative was defined by the addition or subtraction of a concave or convex apex. Chimpanzees were more sensitive to the concave deformation than to the convex deformation. This tendency conforms to the theories of human visual perception that have treated concave features as important factors in reconstructing three-dimensional structures from two dimensional images. Our results suggest that shape representation in chimpanzees is similar to that in humans and that chimpanzees visually process two dimensional images in the same manner as humans. PMID- 17418506 TI - Dynamics of Haemophilus parasuis genotypes in a farm recovered from an outbreak of Glasser's disease. AB - Haemophilus parasuis is a colonizer of the upper respiratory tract of pigs, although it is better known as the etiological agent of Glasser's disease. Interestingly, several strains can be isolated from a single farm, as determined by both genotyping and serotyping. However, it is not known how an outbreak and the subsequent treatment affect the population of H. parasuis strains. In this study, a farm was studied during an outbreak of Glasser's disease and 1 year after antimicrobial treatment and elimination of clinical signs. Bacterial isolation was attempted from nasal swabs and lesions. After isolation, antimicrobial susceptibility, serotype and genotype were determined. Two different genotyping techniques, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus sequence typing (MLST) were used. The H. parasuis strain that was isolated from lesions during the disease outbreak clustered with other virulent strains by both MLST and serotyping analysis. Nasal isolates were included in the corresponding nasal cluster by MLST, but they presented high variability by serotyping. These nasal isolates included serotypes previously classified as virulent and non-virulent. Finally, we found that during the antimicrobial treatment the diversity of strains isolated in the farm was affected and just one strain, which was resistant to the treatment, was detected. One year after the treatment strain diversity was back to normal (three strains). PMID- 17418507 TI - Expression of porcine endogenous retroviruses (PERVs) in melanomas of Munich miniature swine (MMS) Troll. AB - Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pig breeds. Since some of them are able to infect human cells, they might represent a risk for xenotransplantation using pig cells or organs. However, the expression and biological role of PERVs in healthy pigs as well as in porcine tumours is largely unknown. Since we and others have recently shown overexpression of a human endogenous retrovirus, HERV-K, in human melanomas, we studied the expression of PERVs in melanomas of selectively bred Munich miniature swine (MMS) Troll. This breeding herd of MMS Troll is characterised by a high prevalence of melanomas, which histologically resemble various types of cutaneous melanomas in humans. Several genetic factors have been defined when studying inheritance of melanomas and melanocytic nevi in MMS Troll. Here we show that the polytropic PERV-A and PERV-B as well as the ecotropic PERV-C are present in the genome of all melanoma bearing MMS Troll investigated. Most interestingly, in the spleen, but not in other organs, recombinant PERV-A/C proviruses were found. PERV expression was found elevated in melanomas when compared to normal skin and viral proteins were expressed in melanomas and pulmonary metastasis-derived melanoma cell cultures. During passaging of these cells in vitro the expression of PERV mRNA and protein increased and virus particles were released as shown by RT activity in the supernatant and by electron microscopy. Genomic RNA of PERV-A, -B and -C were found in pelleted virus particles. Although PERV expression was elevated in melanomas and pulmonary metastasis-derived cell cultures, the function of the virus in tumour development is still unclear. PMID- 17418508 TI - Quantification of PCV2 capsid transcript in peripheral blood mononuclear cells (PBMCs) in vitro. AB - The presence of PCV2 DNA or spliced capsid mRNA (Cap mRNA) for viral replication was assessed following addition of PCV2 to resting or concanavalin A (ConA) stimulated peripheral blood mononuclear cells (PBMCs). Real-time PCR or real-time RT-PCR assays were used to measure viral DNA or Cap mRNA, respectively. The study demonstrated that PCV2 replication increased in infected PBMCs over time. Replication within infected PBMCs was significantly (P<0.05) increased when PBMCs were stimulated with ConA, compared to unstimulated PBMCs. The data showed a strong correlation between the level of PCV2 Cap mRNA and the level of viral DNA in the ConA stimulated PBMCs. Replication of PCV2 was also assessed in T lymphocyte- and monocyte/macrophage-enriched or monocyte/macrophage-depleted PBMC populations which had been stimulated with ConA for 3 days. It was demonstrated that the enriched T lymphocytes and the monocyte/macrophage-depleted PBMCs had significantly higher Cap mRNA and viral DNA levels (P<0.05) compared to the monocyte/macrophage-enriched population, indicating that in addition to monocytes/macrophages, PCV2 replicates in lymphocytes, particularly T lymphocytes following stimulation. These results suggest that the presence of activated T lymphocytes may play an important role in PCV2 replication and potentially the development of clinical disease. PMID- 17418509 TI - Breed differences in clearance of porcine FSH in hypophysectomized rats. AB - Extracts of anterior pituitary (AP) glands were infused i.v. into hypophysectomized male rats followed by sequential sampling of blood for 120 min. Determination of follicle-stimulating hormone (FSH) concentrations established that FSH from Chinese Meishan males decreased in the circulation of rats more slowly than FSH in extracts of AP from crossbred occidental pigs (P<0.003). Additionally, FSH from AP extracts of castrated males disappeared somewhat more slowly (P<0.06) than FSH from extracts of boars. Evaluation of FSH by bioassay and radioimmunoassay yielded similar concentrations in AP from Meishan and crossbred boars. Serum testosterone concentrations increased with time through 90 min after infusion of AP, but the rate of increase of testosterone was not related to amount of luteinizing hormone (LH) that was administered indicating LH receptor saturation. Unexpectedly, the rate of increase in testosterone was more rapid with AP extracts from boars than with extracts from castrated males. Observations from the current study imply structural alterations of FSH in the AP of Meishan males relative to crossbred males allowing sustained concentrations in the circulation, and this FSH possesses similar activation of the FSH receptor. The amount of LH in the AP extracts saturated the LH receptors of the hypophysectomized male rats, but some factor in extracts of boars differed from those of castrated males. PMID- 17418510 TI - Effect of alkylcarbonates of gamma-cyclodextrins with different chain lengths on drug complexation and release characteristics. AB - Alkylcarbonates of gamma-cyclodextrins were produced and their inclusion complexes with four poorly water-soluble drugs of different structures and solubilities were prepared. The alkylcarbonates and the alkylcarbonate drug complexes were characterized by DSC and XRPD; the physical mixtures were used as control. Solubility capacities were evaluated by phase solubility studies. The effect of alkyl chain length on the complexation and release behaviour was investigated as well. The XRPD patterns of alkylcarbonates showed that the derivatives lose the original crystallinity of gamma-cyclodextrins. The series of alkylcarbonates formed inclusion complexes with the drugs considered. Both XRPD and DSC analyses did not show neither the reflections of the crystalline structures nor the melting peaks of the drugs, respectively. These gamma cyclodextrin derivatives can improve drug solubility and influence the drug release rates while the alkyl chain length may affect these properties. PMID- 17418511 TI - In vivo trypanocidal activity of Annona senegalensis Pers. leaf extract against Trypanosoma brucei brucei. AB - Chemotherapy of African trypanosomiasis still remains far from being satisfactory. There is the urgent need for therapeutic agents that are effective, affordable and accessible to the rural poor in Africa who bear most of the disease burden. Root preparations of Annona senegalensis Pers. is claimed by traditional medicine practitioners to be effective in the treatment of sleeping sickness. Validation of this claim, evaluation of the therapeutic effects of other parts of the plant, and standardization of the preparations are necessary in order to fully exploit the chemotherapeutic potentials of this plant. We have evaluated the chemotherapeutic effects of extracts of the leaves, whole root, root and stem bark of the plant in experimental trypanosomiasis. Crude and partially purified aqueous extracts of the leaves, at a dose of 200mg/kg body weight per day completely cured experimental Trypanosoma brucei brucei infection in mice. Sub-inoculation of blood and cerebrospinal fluid drawn from the cured mice into healthy mice failed to produce any infection within 60 days of post inoculation. Pre-treatment of healthy mice with the crude extract before infection did not prevent establishment of infection. Administration of 5000mg/kg body weight of the crude extract did not lead to fatality in mice. Preliminary phytochemical screening showed the presence of tannin, phlobatanin, and saponin. PMID- 17418512 TI - Regulatory effects of Codonopsis lanceolata on macrophage-mediated immune responses. AB - Codonopsis lanceolata L. has long been used as a folk medicine in Korea, Japan and China for the treatment of lung inflammatory diseases. In this study, therefore, we aimed to demonstrate its ethnopharmacological activity by examining macrophage-function regulating effects. The total methanol extracts of fresh leaves (l-TME) or roots (r-TME) of Codonopsis lanceolata L. significantly suppressed the production of pro-inflammatory mediators (nitric oxide [NO] and tumor necrosis factor [TNF-alpha]) without altering mRNA levels. The expression of interleukin (IL)-3 and IL-6, however, was strongly diminished. According to the analysis of signaling enzyme activation by immunoblotting, phospho-IkappaB levels, a representative pro-inflammatory gene activation pathway, were not affected by the TMEs. By contrast, the Raf-ERK signaling pathway, which was involved in regulation of post-translational modification of pro-inflammatory gene products, was strongly blocked after 6-h of exposure. Moreover, l-TME down regulated LPS-mediated phagocytic uptake and CD29-mediated cell-cell adhesion, while r-TME strongly up-regulated these two cellular events as well as fibronectin-cell adhesion. The surface levels of the costimulatory molecules (CD80 and CD86) of RAW264.7 cells were also enhanced by these extracts. l-TME also diminished functional activation (assessed by NO production) and the surface level of dectin-1, but not toll-like receptor (TLR)-2. Taken together, these data suggest that Codonopsis lanceolata may have the ability to modulate macrophage mediated immune responses, thus contributing to its anti-inflammatory activity. PMID- 17418513 TI - Pubis growth study: applicability in sexual and age diagnostic. AB - The growth of five variables of the ischiopubic area was analyzed from bone material from birth to old age. The main purpose was to evaluate its significance and capacity for age and sex determination during and after growth. The material used consisted of 327 specimens from four documented Western European collections. Growth curves were calculated by polynomial regression for two classical variables of the ischiopubic area (pubis length and ischiopubic index) and three new variables of the pubic acetabular area (horizontal and vertical diameter of the pubic acetabular area and the pubic acetabular index). None of the curves showed lineal growth, with the exception of the ischiopubic index and the masculine vertical diameter of the pubis acetabular area. Pubis length has the most complicated growth, expressed by a five-degree polynomial. All the variables are useful for adult sex determination, except the pubic acetabular index. The ischopubic index, vertical diameter of the pubic acetabular area and the pubic acetabular index seem to be good variables for sub-adult sex determination. For age estimation the best variables, in both archaeological and forensic remains, are the absolute measurements (pubic length, vertical and horizontal diameter of the pubis). However, pubis length is the best variable for age estimation because it can be applied until 25 years of age. PMID- 17418514 TI - Doped hydrophobic silica nano- and micro-particles as novel agents for developing latent fingerprints. AB - Novel hydrophobic silica based particles have been developed to visualise latent fingerprints. The composition of the particles has been designed to maximise both hydrophobic and ionic interactions between a variety of coloured and fluorescent reporter molecules and the silicate backbone within the particles. The resulting doped particles retain the incorporated dyes with high affinity. In addition, a variety of sub-particles have also been embedded to again produce coloured or magnetisable hydrophobic particles. The particles can be harvested as nanoparticles or microparticles. The former are applied to latent fingerprints as an aqueous suspension and the latter as a dusting agent using brushes or a magnetic wand. Examples of the prints produced using these agents are given. The resulting prints have good definition. PMID- 17418515 TI - A case study on the application of isotope ratio mass spectrometry (IRMS) in determining the provenance of a rock used in an alleged nickel switching incident. AB - The application of isotope ratio mass spectrometry (IRMS) in forensic science to establish the provenance of a range of questioned substances including soils, drugs, explosives, currency, ivory and rhino horn has been widely documented. The present study wishes to highlight the applicability of IRMS and specifically stable carbon IRMS in determining the provenance of a carbonate rock that was switched for nickel metal exported from South Africa to Israel. The technique employed effectively argued against a South African origin for the rock whilst simultaneously supporting an Israeli origin, enabling investigators to focus their attention accordingly. The study represents the first documented instance known to the authors where IRMS has been employed in the forensic geo-location of a rock. PMID- 17418516 TI - Early dietary treatments with Lorenzo's oil and docosahexaenoic acid for neurological development in a case with Zellweger syndrome. AB - We treated a girl with Zellweger syndrome using a special infant formula supplemented with middle chain triglyceride (MCT) milk, docosahexaenoic acid (DHA), Lorenzo's oil, and Lunaria oil, which is rich in nervonic acid (C24:1). We examined the fatty acid contents of the plasma and red blood cell (RBC) membrane. Neurological development was evaluated using Denver developmental screening test and auditory brainstem response (ABR). Her delayed neurological development, liver dysfunction, and cholestasis were all improved 2 weeks after starting the dietary treatment. DHA level in RBC membranes was increased and very long chain fatty acid (VLCFA,C26:0) levels were decreased. Our findings suggest that the dietary treatment with combination of MCT milk, DHA, Lorenzo's oil, and Lunaria oil in the patients with Zellweger syndrome bring some benefits for neurological development. PMID- 17418517 TI - Quantification of missing and overlapping data in multiple breath hold abdominal imaging. AB - Magnetic resonance imaging (MRI) of the abdomen is often performed in multiple breath holds which are designed to contiguously cover the region of interest. This technique may result in a failure to image all the appropriate area, and the extent of this failure is difficult to appreciate on a set of 2D slices. With reference to three patient cases, we present a method to quantify the extent of this problem and suggest a solution. First, we manually delineate the region of interest on a single breath hold fast spoiled gradient echo (FSPGR) sequence. Subsequently, we align images acquired in separate breath holds to this reference volume. A coloured 3D presentation makes the extent of unimaged and repeatedly imaged areas clearly visible to the clinician. The alignment also helps radiologists to accurately determine the location of individual slices. The described method can easily be automated and is ideally implemented at the scanner console, ensuring the availability of contiguously sampled datasets to radiologists with minimum user interaction from the radiographer. Such datasets enable the deployment of robust 3D analysis algorithms. PMID- 17418518 TI - The effect of varying echo spacing within a multiecho acquisition: better characterization of long T2 components. AB - A 48-echo pulse sequence with five different echo-spacing combinations was examined to determine how one can most effectively measure the T2 relaxation characteristics of cerebral tissue containing a long T2 component. For each scan, the first 32 echoes had an echo spacing of 10 ms, while the spacing for Echoes 33 48 (DeltaTE2) was 10, 20, 30, 40 or 50 ms. In an in vivo study using 10 normal volunteers, it was found that the resolution of T2 distribution peaks for both myelin water (approximately 20 ms) and intracellular/extracellular (IE) water (approximately 80 ms) improved as DeltaTE2 increased. The geometric mean T2 values of the main peak agreed within the error for all DeltaTE2 values. A phantom study simulated T2 relaxation distributions that are expected in the brains of patients with demyelinating diseases. For phantoms in which the T2 values of the IE and lesion (200-500 ms) water compartments were separated by at least a factor of 3, each compartment in the distribution was better resolved when DeltaTE2=40 or 50 ms. On the basis of these results, we recommend the use of extended DeltaTE2 values for imaging patients with lesions, without the risk of losing valuable short T2 information. PMID- 17418519 TI - Fast multivoxel two-dimensional spectroscopic imaging at 3 T. AB - The utility of multivoxel two-dimensional chemical shift imaging in the clinical environment will ultimately be determined by the imaging time and the metabolite peaks that can be detected. Different k-space sampling schemes can be characterized by their minimum required imaging time. The use of spiral-based readout gradients effectively reduces the minimum scan time required due to simultaneous data acquisition in three k-space dimensions (k(x), k(y) and k(f(2))). A 3-T spiral-based multivoxel two-dimensional spectroscopic imaging sequence using the PRESS excitation scheme was implemented. Good performance was demonstrated by acquiring preliminary in vivo data for applications, including brain glutamate imaging, metabolite T(2) quantification and high-spatial resolution prostate spectroscopic imaging. All protocols were designed to acquire data within a 17-min scan time at a field strength of 3 T. PMID- 17418520 TI - Simple phase method for measurement of magnetic field gradient waveforms. AB - Magnetic field gradients play a fundamental role in MR imaging and localized spectroscopy. The MRI experiment, in particular fast MRI, relies on precise gradient switching, which has become more demanding with the constantly growing number of fast imaging techniques. Here we present a simple MR method to measure the behavior of a magnetic field gradient waveform in an MR scanner. The method employs excitation of a thin slice, followed by application of the studied gradient and simultaneous FID acquisition. Measurements of different gradient waveforms were performed with a spherical phantom filled with doped water and positioned at the isocenter of the gradient set. The presented experiments demonstrate the capability of the technique to measure different gradient waveforms with an estimated error of less than 200 microT/m. PMID- 17418521 TI - LC-ESI-MS method for the determination of bisoprolol in human plasma. AB - A sensitive liquid chromatography-electrospray ionization-mass spectrometry (LC ESI-MS) method has been developed and validated for the determination of bisoprolol in human plasma, using metoprolol as internal standard (I.S.). After alkalization with sodium hydroxide, the samples were extracted with ethyl acetate and separated by HPLC on a ZORBAX SB-C18 column with a mobile phase of 10 mM ammonium acetate buffer containing 0.1% formic acid-methanol (32:68, v/v) at a flow rate of 1 ml/min. The chromatographic separation was achieved in less than 5 min. The linearity was established over the concentration range of 0.05-120 ng/ml. The intra- and inter-run standard deviation was less than 3.8 and 7.5%, respectively. The method had been successfully applied to study the relative bioavailability of bisoprolol fumarate tablets in healthy Chinese volunteers. The pharmacokinetic parameters of the reference and test tablets have been compared. PMID- 17418522 TI - Social marketing meets health literacy: Innovative improvement of health care providers' comfort with patient interaction. AB - OBJECTIVE: It is essential to train health care providers to deliver care sensitive to the needs of diverse individuals with varying degrees of health literacy. We aimed to evaluate an innovative, theory-based, educational intervention involving social marketing and health literacy. METHODS: In 2006 at a large medical school, all first-year students were exposed to the intervention. They completed pre- and post-test anonymous surveys including demographic data, covariates, and key outcome variables. Paired t-tests and multiple linear regression were used to evaluate the intervention and to determine independent associations among the key outcome variables. RESULTS: Post-intervention scores were significantly higher than pre-intervention scores for social marketing (3.31 versus 1.90, p<0.001), health literacy (3.41 versus 2.98, p<0.001), and comfort in brochure development (3.11 versus 2.52, p<0.001) (N=83). After controlling for demographic and covariate data, health literacy and comfort in brochure development were independent predictors of comfort interacting with diverse populations. CONCLUSION: A brief intervention involving social marketing and health literacy can improve skills that improve medical students' comfort with patients of diverse backgrounds. PRACTICE IMPLICATIONS: Health care providers can be taught educational principles and skills involved in developing effective patient education materials. These skills may improve providers' comfort with direct patient interaction. PMID- 17418523 TI - [Does the parapenylenediamine cross the placenta?]. PMID- 17418524 TI - One Argonaute family member, Eif2c2 (Ago2), is essential for development and appears not to be involved in DNA methylation. AB - To elucidate the epigenetic role of RNAi in mammals, we disrupted the gene for Eif2c2 (Ago2), which works as the sole slicer of RNAi in the Argonaute family. In mice, disruption of Eif2c2 leads to embryonic lethality early in development after the implantation stage. This phenotype is completely different from that in a previous report, but somewhat similar to the disruption of Dicer1, another important component of RNAi. We also show that Eif2c2 is not required for the maintenance of DNA methylation in imprinted genes, centromeric repeats, and Xist. This suggests that developmental defects in the Eif2c2-deficient mouse are caused not at the transcriptional level, but rather at the posttranscriptional level through the miRNA-protein complex. PMID- 17418525 TI - Effects of octamethylcyclotetrasiloxane (D4) on the luteinizing hormone (LH) surge and levels of various reproductive hormones in female Sprague-Dawley rats. AB - The objectives of this study were to assess the potential for D(4) to suppress the pre-ovulatory lutenizing hormone (LH) surge, to block or delay ovulation, and to evaluate potential effects on reproductive hormones in rats. Female Sprague Dawley Crl:CD (SD) IGS BR rats received whole-body vapor inhalation exposure to D(4) (0, 700, or 900ppm) 6h per day for 3 days. Trunk blood obtained on proestrus at 10a.m. was evaluated for levels of follicle stimulating hormone (FSH), estradiol (E2), estrone (E1), and progesterone (P4). Other rats had serial blood samples collected via cannula at 2, 4, 6, 8, and 10p.m. on the day of proestrus and plasma evaluated for LH and prolactin (PRL). Trunk blood was collected at 8a.m. of estrus and plasma evaluated for FSH, E2, E1, and P4. At 10a.m. on proestrus, significant increases in E1 levels in the 700 and 900ppm groups and significant increases in P4 levels in the 900ppm group were noted. At 8a.m. on estrus, significant increases in E1, E2, in the E1/E2 ratio and decreases in FSH were noted in the 700 and 900ppm groups. The major effect on the LH profile was observed most clearly when the rats were grouped by ovulatory status, animals that did or did not ovulate. Regardless of treatment, suppression of the LH surge correlated with blocked ovulation. The percentage of rats that ovulated was (700ppm, 42%; 900ppm, 31%) compared to controls (79%). Overall, the data indicate that high exposures to D(4) attenuated the pre-ovulatory LH surge and significantly decreased the portion of female rats that ovulated. PMID- 17418526 TI - Moderating influence of the drinking water disinfection by-product dibromoacetic acid on a dithiocarbamate-induced suppression of the luteinizing hormone surge in female rats. AB - The disinfection by-product dibromoacetic acid (DBA) has been found in female rats to increase circulating concentrations of both estradiol (E2) and estrone (E1). This effect is apparently due, at least in part, to a suppression in hepatic catabolism. The present study investigated whether DBA, by increasing sex steroid levels, is able either to augment the hypothalamic up-regulation involved in triggering a luteinizing hormone (LH) surge, or to affect the ability of the neurotoxicant sodium dimethyldithiocarbamate (DMDC) to block the surge. Sprague Dawley rats were gavaged for 14 days with DBA (0-150mg/kg) and ovariectomized on dosing day 11, and at the same time implanted with an estradiol capsule to generate daily LH surges. An injection of 0.1mM/kg DMDC was administered at 13:00h on day 14 and blood was sampled over the afternoon. DBA induced a dose related increase in total estrogens. For identified surges, areas under the LH curve partitioned into two groups, comprising the two lower (0 and 37.5mg/kg DBA) and the two higher (75 and 150mg/kg) treatment groups. Consequently, low and high DBA groups were compared and found to be significantly different. At 150mg DBA/0.1mM DMDC, the timing of an identifiable LH peak was comparable to non-DMDC females, unlike the 37.5mg DBA/0.1mM DMDC group in which the appearance of peak concentrations was delayed. A significant effect with DBA treatment alone was not present. Results indicated that this exposure to DBA induced a dose-related increase in total estrogen concentrations that paralleled a diminished DMDC blockade of the LH surge. The effect appeared to be attributable to an augmentation in the estrogen-associated up-regulation in brain mechanisms stimulating the surge. PMID- 17418527 TI - CAD in clinical trials: current role and architectural requirements. AB - Computer-aided diagnosis (CAD) technology is becoming an important tool to assess treatment response in clinical trials. However, CAD software alone is not sufficient to conduct an imaging-based clinical trial. There are a number of architectural requirements such as image receive (from multiple field sites), a database for storing quantitative measures, and data mining and reporting capabilities. In this paper we describe the architectural requirements to incorporate CAD into clinical trials and illustrate their functionality in therapeutic trials for emphysema. PMID- 17418528 TI - Pharmacogenomics and its implications for autoimmune disease. AB - A striking failure of modern medicine is the debilitating and lethal consequences of adverse drug reactions (ADRs) which rank as one of the top ten leading causes of death and illness in the developed world with direct medical costs of 137-177 billion US dollars annually in the USA. Although many factors influence the effect of medications (i.e. age, organ function, drug interactions), genetic factors account for 20-95% of drug response variability and play a significant role in the incidence and severity of ADRs. The field of pharmacogenomics seeks to identify genetic factors responsible for individual differences in drug efficacy and adverse drug reactions. Pharmacogenomics has led to several genetic tests that provide clinical dosing recommendations. For autoimmune disease, pharmacogenomics has led to several DNA-based tests to improve drug selection, optimize dosing, and minimize the risk of toxicity. The 'GATC' project is a nation-wide project established in Canada to identify novel predictive genomic markers of severe ADRs in children. An ADR surveillance network has been established in all of Canada's major children's hospitals, serving up to 75% of all Canadian children. The goal of the project is to identify patients experiencing specific ADRs, collect DNA samples, and apply genomics-based technologies to identify ADR-associated genetic markers. PMID- 17418529 TI - The regulatory T cell gene FOXP3 and genetic susceptibility to thyroid autoimmunity: an association analysis in Caucasian and Japanese cohorts. AB - FOXP3 is a key gene in the development of regulatory T cells (Treg). FOXP3 expression commits naive T cells to become Treg cells. Indeed, mutations in the FOXP3 gene cause severe systemic autoimmune diseases in humans and in mice. Therefore, we hypothesized that the FOXP3 gene may be associated with thyroid autoimmunity which is among the typical autoimmune diseases that develop in individuals with FOXP3 mutations. Moreover, the FOXP3 gene is located within an X chromosome locus (Xp11.23) previously shown to be linked with autoimmune thyroid diseases (AITD). We tested the FOXP3 gene locus for association with AITD in two large cohorts of US Caucasians and Japanese AITD patients. We analyzed 269 Caucasian AITD patients (52 males and 217 females) and 357 Caucasian controls (159 males and 198 females), as well as 377 female Japanese AITD patients and 179 female Japanese controls. The FOXP3 gene locus was analyzed using four microsatellite polymorphisms [(GT)n; (TC)n; DXS573; DXS1208] flanking the FOXP3 gene locus. Interestingly, while no association was found between FOXP3 polymorphisms and AITD in the Japanese cohort there was a significant association in the Caucasian cohort. There was a significant association of the (TC)n polymorphism with AITD in the Caucasian male AITD patients (p=0.011; 5 degrees of freedom [df]). Similarly, there was an association between the DXS573 microsatellite and AITD in the Caucasian female AITD patients (p=0.00023; 4 df). These results suggest that polymorphisms of the FOXP3 gene may play a role in the genetic susceptibility to AITD in Caucasians, perhaps by altering FOXP3 function and/or expression. PMID- 17418530 TI - Reliability of sentinel lymph-node extirpation as a diagnostic method for malignant melanoma of the head and neck region. AB - A series of 106 patients with malignant melanoma of the head and neck and clinically negative local lymph-node status were included in a multimodal therapy programme and underwent sentinel lymph-node extirpation in 1999-2003. Out of 246 preoperatively marked lymph nodes, only 172 (70%) were identified intraoperatively and removed. In 89% of all patients at least one sentinel lymph node was removed. Histological examination revealed metastases in the sentinel lymph nodes of 17 patients. In the mean follow-up period of 47 months (range 4-76 months), regional lymph-node metastases were found in another eight patients. The non-marked lymph nodes that were often removed at the same time, in an elective cervical lymph-node dissection, did not reveal any metastasis in any of the cases where the sentinel lymph nodes were negative. The sensitivity of sentinel lymph node extirpation was influenced by the length of the follow-up period and the detection rate, and was 68% (17/17+8), a result superior to that of any other diagnostic tool. Sentinel lymph-node extirpation is a valuable method in addition to elective lymph-node dissection. PMID- 17418531 TI - A new method of monitoring osteomyelitis. AB - Chronic infections of bone such as osteomyelitis are frequent events, especially in immunocompromised or diabetic patients, and costly on a national level. Incorrect treatment or delayed diagnosis may lead to loss of the affected extremity or mandible. The aim of this study was to assess the possible value of urinary lysylpyridinoline (LP) and hydroxylysylpyridinoline (HP) concentrations in the monitoring of mandibular osteomyelitis. Patients were assigned to the following groups: group 1 (n=85), control; group 2a (n=38), patients with active disease; group 2b (n=25), patients of group 2a 6 months after successful treatment; group 2c (n=7), patients of group 2a with ongoing osteomyelitis 6 months after treatment. The range and upper limit of normal values (HP(max) and LP(max)) were determined in group 1. Levels of LP and HP were measured by high performance liquid chromatography and fluorescence detection. There was a significant decrease (mean 45.43% for HP and 32.12% for LP) in samples of group 2b compared to 2a (P<0.001 for HP and LP). There was a significant increase in HP values in samples from group 2c compared to 2a (P=0.018). The urinary concentrations of HP and LP appear to act as a marker of disease activity, with a decrease reflecting treatment success and an increase or stable values indicating persistent disease. An inexpensive tool (US$5 per analysis) for the monitoring of osteomyelitis is described. PMID- 17418532 TI - Repair of a laryngeal fracture using miniplates. AB - Injuries to the larynx (voice box) can lead to loss of vital functions; the airway may be obstructed, the voice distorted or lost, and the protection of the airway during swallowing may fail. In order to preserve these functions, a stable repair that restores the anatomy as closely as possible is needed. The repair should interfere minimally with the neuromuscular functions of the larynx. The case is described of a 59-year-old male who suffered a severe laryngeal fracture in a work-place accident. Utilizing miniplates, the fracture was reduced and fixed in the correct position. After healing, the patient could be decannulated and has regained his voice, swallows without difficulty and has a patent airway. The results have been sustained over a 2-year follow-up. PMID- 17418533 TI - Fixed partial dentures on two implants: raising comfort in irradiated edentulous patients. AB - Surgical treatment of malignancies involving the oral cavity often results in an altered anatomical situation, which may severely hamper oral function. Postoperative radiotherapy resulting in reduced salivary secretion further worsens functions such as speech, chewing (mastication) and swallowing. In order to improve oral functioning, the use of dental implants was proposed. To reconstruct the edentulous mandible, especially in cases involving postoperative irradiation, implant-borne prosthetic construction on four permucosal implants has been advocated, but this treatment concept has some practical drawbacks. As the chewing force is reduced as a result of irradiation of the masticatory muscles, it is hypothesized that a fixed prosthetic construction on only two implants is a feasible alternative. Up until now 10 patients have been treated successfully according to this concept at the authors' clinic. PMID- 17418534 TI - Learning by doing virtually. AB - Selective reduction of bone without collateral damage (nerves, teeth) is essential in apicectomy. To test whether skills acquired on a virtual apicectomy simulator (VOXEL-MAN system with integrated force-feedback) are transferable from virtual to physical reality, two groups of trainees were compared. Group 1 received computer-based virtual surgical training before performing an apicectomy in a pig cadaver model. The probability of preserving vital neighboring structures was improved significantly, i.e. six-fold, after virtual surgical training (P<0.001). The average volume of the bony defects created by the trainees of Group 2 (mean: 0.47 ml) was significantly (P<0.001) larger than by the trainees of Group 1 (mean: 0.25 ml). Most importantly, the ability to objectively self-assess performance was significantly improved after virtual training. Training with a virtual apicectomy simulator appears to be effective, and the skills acquired are transferable to physical reality. PMID- 17418535 TI - Isolated hypoglossal nerve palsy due to a vascular anomaly. AB - The case of a patient with an unusual cause of an isolated hypoglossal nerve palsy is presented. Magnetic resonance (MR) imaging demonstrated marked dilatation and ectasia of the right vertebral artery and basilar artery, thereby causing compression of the brainstem. It is therefore proposed that a dilated vascular loop compression of the nerve rootlets, as they exit the medulla, has caused the hypoglossal nerve palsy. This case demonstrates that excellent images can be obtained from MR imaging, and also emphasises that other pathologies must be considered in the differential diagnosis of hypoglossal nerve palsy. PMID- 17418536 TI - Promotion of mindfulness in psychotherapists in training: preliminary study. AB - This study examined whether the promotion of mindfulness in psychotherapists in training can influence the treatment results of their patients. The therapeutic course and treatment results of 196 inpatients, who were treated during a nine week period by nine psychotherapists in training, were compared: in the first phase of the study, the treatment group without (CG, historical control group, n=55), and in the second phase the treatment group with, (MFG, n=58) therapists who were currently practicing Zen meditation. The results of treatment were examined (according to the intent-to-treat principle) with the Session Questionnaire for General and Differential Individual Psychotherapy (STEP), the Questionnaire of Changes in Experience and Behaviour (VEV) and the Symptom Checklist (SCL-90-R), and showed significantly better results in the MFG. PMID- 17418537 TI - A meta-analysis of clinical trials comparing the serotonin (5HT)-2 receptor antagonists trazodone and nefazodone with selective serotonin reuptake inhibitors for the treatment of major depressive disorder. AB - OBJECTIVE: To compare response rates among patients with major depressive disorder (MDD) treated with either a serotonin-2 (5HT2-) receptor antagonist or a selective serotonin reuptake inhibitor (SSRI). METHODS: Medline and PubMed were searched for double-blind, randomized clinical trials comparing either trazodone or nefazodone with an SSRI for the treatment of MDD. Data from 9 reports involving a total 988 patients were identified and combined using a random effects model. RESULTS: Patients randomized to treatment with a 5HT2 antagonist were as likely to experience clinical response as patients randomized to treatment with an SSRI (RR=1.002, 95% CI: 0.85-1.17, P=0.978). Pooled response rates for trazodone/nefazodone and the SSRIs were 61.1% and 61.7%, respectively. There was also no difference in overall discontinuation rates (P=0.334), discontinuation due to adverse events (P=0.676), or discontinuation due to inefficacy (P=0.289) between the two groups. CONCLUSIONS: These results suggest that the 5HT2-receptor antagonists trazodone and nefazodone and the SSRIs do not differ with respect to their overall efficacy and tolerability in the treatment of MDD. Although the sample size was relatively large and conveyed sufficient statistical power to test for differences in the overall sample, depression is a heterogeneous condition and differences may exist between treatments in particular subgroups of patients. PMID- 17418538 TI - Economic consequences of alternative medication strategies in first episode non affective psychosis. AB - BACKGROUND: Maintenance treatment appears to be successful in preventing relapses in first episode psychosis, but is also associated with side effects. Guided discontinuation strategy is a less intrusive intervention, but may lead to more relapses. In the current economic evaluation, costs and health outcomes of discontinuation strategy will be compared with the results of maintenance treatment in patients with remitted first episode psychosis. METHOD: The study was designed as a randomised clinical trial. In total 128 patients were prospectively followed for 18 months after six months of stable remission. The economic evaluation was conducted from a societal perspective. Quality-adjusted life years (QALYs) were used as primary health outcome in the economic evaluation. Relapse rates were assessed in addition to various other secondary outcomes. RESULTS: There were no relevant differences in mean costs between groups during the study. Total costs were largely influenced by costs related to admissions to psychiatric hospitals. No differences between groups were found for QALY results. CONCLUSIONS: There were no indications that either of the examined interventions is superior to the other in terms of costs or QALY results. Additional results indicated that the relapse rate in discontinuation strategy was twice as high, but without an increase in hospital admissions or negative consequences on other clinical outcomes. For a minority of remitted first episode patients, guided discontinuation strategy may offer a feasible alternative to maintenance treatment. PMID- 17418539 TI - Simulations of molecular dynamics in solid-state NMR spectra of spin-1 nuclei including effects of CSA- and EFG-terms up to second order. AB - By numerical simulations MAS and QCPMG methods for acquiring spectra of spin-1 nuclei were compared in order to determine the most sensitive experiment for analysis of molecular dynamics. To comply with the large quadrupolar constants for 14N and the CSA reported for 6Li both of these interactions are included up to second order. For 2H and 6Li both QCPMG and single-pulse MAS experiments were suitable for dynamics studies whereas the single-pulse MAS experiment were the method of choice for investigation of 14N dynamics for C(Q)'s larger than 750kHz at 14.1T. This property prohibits excitation of the 14N lineshape using either single hard or softer composite rf-pulses. Focusing on 14N it was demonstrated that the centerband lineshape is sensitive toward both off-MAS and CSA effects. In addition, excitation by real-time pulses showed that proper lineshapes corresponding to a site with a C(Q) of 3MHz may be excited by a very short pulse. PMID- 17418540 TI - Investigations of Li-containing SiCN(O) ceramics via 7Li MAS NMR. AB - Lithium-containing silicon (oxy)carbonitride ceramics (SiCN(O):Li) were synthesized via precursor-to-ceramic-transformation of Li-containing (poly)silazanes. The precursors were obtained by lithiation of 2,4,6-trimethyl 2,4,6-trivinylcyclotrisilazane with n-butyllithium and by reaction of a commercial poly(organosilazane) VL20 with metallic lithium. The annealing treatment was carried out at temperatures between 200 and 1400 degrees C in argon (DeltaT=200 degrees C) and yielded Li-containing silicon (oxy)carbonitride. X-ray powder diffraction revealed that the resulting SiCN(O):Li ceramics were basically amorphous up to temperatures of 1000 degrees C and formed LiSi(2)N(3), graphite and silicon carbide as crystalline phases at higher temperatures. (7)Li MAS NMR spectroscopy was carried out to investigate the structure of the Li-containing phases and to study the reaction path of metallic Li with polysilazane. Based on the NMR spectra, there is almost no difference found in the chemical shift of the SiCN(O):Li ceramics obtained at different temperatures. Accordingly, Li is assigned to be mainly coordinated to N and O present as contaminant element. Relaxation time measurements showed that the most mobile Li(+) species seems to be present in the product obtained in the pyrolysis temperature range between 600 and 1000 degrees C. PMID- 17418541 TI - Fabrication of ultrathin polyelectrolyte fibers and their controlled release properties. AB - Ultrathin fibers comprising 2-weak polyelectrolytes, poly(acrylic acid) (PAA) and poly(allylamine hydrochloride) (PAH) were fabricated using the electrospinning technique. Methylene blue (MB) was used as a model drug to evaluate the potential application of the fibers for drug delivery. The release of MB was controlled in a nonbuffered medium by changing the pH of the solution. The sustained release of MB in a phosphate buffered saline (PBS) solution was achieved by constructing perfluorosilane networks on the fiber surfaces as capping layers. Temperature controlled release of MB was obtained by depositing temperature sensitive PAA/poly(N-isopropylacrylamide) (PNIPAAM) multilayers onto the fiber surfaces. The controlled release of drugs from electrospun fibers have potential applications as drug carriers in biomedical science. PMID- 17418542 TI - Interaction and incorporation of ovalbumin with stearic acid monolayer: Langmuir Blodgett film formation and deposition. AB - In this communication, the surface activity of the ovalbumin (OVA) at the air/water interface was studied to establish the nature of the interaction with the stearic acid (SA) monolayer, based on Langmuir-Blodgett (LB) technique. The interaction was monitored by studying the time (t) variation of surface pressure (pi) at constant area (A). The growth of pi with time indicates a positive association between the SA and the OVA molecules. The surface compressibility analysis has been performed to specify the phase transition of OVA-SA mixed monolayer. Incorporation/association of OVA within the SA monolayer led to noteworthy changes in surface compressibility and was surface pressure as well as protein concentration dependent. Both the hydrophobic and the Vander wall type interactions are found to be responsible for the association. The quenching of tyrosine band in tryptophan excitation spectrum is observed in steady-state fluorescence spectroscopy. This suggests that the tyrosine is the probable binding site with SA. Due to incorporation of SA, the energy transfer from tyrosine to tryptophan is hindered. At higher pressure, OVA tend to squeeze out from the SA monolayer. The high-resolution field emission scanning electron microscope (FE-SEM) image confirms this observation. Aggregated protein structure observed at high pressure indicates unfolding of protein. PMID- 17418543 TI - Morphological observations on a lipid-based drug delivery system during in vitro digestion. AB - The in vitro digestion of a self nano-emulsifying drug delivery system (SNEDDS) was visualized by cryogenic transmission electron microscopy (Cryo-TEM). The dynamic lipolysis model, simulating the environment of the gastrointestinal tract in fasted conditions, was used for this purpose. The results revealed that micelles are present during the entire lipolysis process. Oil droplets from the self nano-emulsifying drug delivery system are transformed to spherical or elongated unilamellar vesicles as lipolysis progresses. Low numbers of bilamellar and open vesicles were detected. After 50% hydrolysis a decrease in the number of unilamellar vesicles and oil droplets was observed. Furthermore, the electrical properties of the oil droplets were investigated by measuring their zeta potential values as a function of time. An increase (in absolute values) to the zeta-potential of the hydrolyzing SNEDDS droplets observed versus time implying (binding or incorporation) of the micelles to the surface. The current data emphasize that Cryo-TEM combined with the in vitro dynamic lipolysis model can offer useful information on the formation of the various colloid phases during in vitro digestion of lipid-based formulations. Furthermore, it can provide a better understanding of the in vivo behavior of these systems, as well the solubilization of lipophilic drug compounds, offering new insights for designing and optimizing oral lipid-based formulations and possibly predicting their in vivo behavior. Such methodology can be a useful tool for the strategic development of lipid-based formulations. PMID- 17418544 TI - [Procalcitonin bedside testing in the pediatric emergency department]. PMID- 17418545 TI - [RAS signalling pathway and its syndromes]. PMID- 17418546 TI - [Consequences of PTH resistance on adult bone]. PMID- 17418547 TI - [Increased incidence of invasive meningococcal disease in Seine-Maritime. The evolving epidemiology due to the B:14:P1.7,16 strain]. PMID- 17418548 TI - Adhesion forces in interactive mixtures for dry powder inhalers--evaluation of a new measuring method. AB - Dry powder inhalers mostly contain carrier based formulations where micronized drug particles are adhered to coarse carrier particles. The performance of the dry powder inhaler depends on the inhaler device, the inhalation manoeuvre and the formulation. The most important factor influencing the behaviour of the formulation is the adhesion force acting between the active ingredient and the carrier particles, which can be measured using different methods, for example the centrifuge technique or atomic force microscopy. In this study the tensile strength method, usually applied to determine cohesion forces between powder particles of one material, is optimized for adhesion force measurements between powder particles of unlike materials. Adhesion force measurements between the carrier materials lactose or mannitol and the drug substance salbutamol sulphate using the tensile strength method and the atomic force microscopy show higher values with increasing relative humidity. Consequently, the fine particle fraction determined using the Next Generation Impactor decreases with increasing relative humidity as a result of the enhanced interparticle interactions. PMID- 17418549 TI - Preparation and characterization of a potent, long-lasting recombinant human serum albumin-interferon-alpha2b fusion protein expressed in Pichia pastoris. AB - A long-lasting recombinant human serum albumin-interferon-alpha2b fusion protein (rHSA/IFNalpha2b) was prepared and its structure and biological activities were studied. rHSA/IFNalpha2b was expressed in methylotrophic yeast Pichia pastoris with HSA's natural signal peptide and purified by dye affinity chromatography, hydrophobic interaction chromatography, ion exchange chromatography and Sephadex G25. Purity of the prepared rHSA/IFNalpha2b was greater than 97% analyzed by non reduced SDS-PAGE and RP-HPLC. Structure and biological activities of the prepared rHSA/IFNalpha2b were characterized by physical, chemical and biological methods. Its pI was 5.3 and showed a single band on IEF gel. Molecular weight determined by MALDI-TOF was 86004.3+/-29.2. Amino-terminal and carboxyl-terminal amino acid sequences were identical to predicted sequence. Its specific activity in vitro was 6.3+/-0.8x10(5) IU/mg fusion protein, retaining about 1.4% of that of unmodified rIFNalpha on a molar basis. After administered in monkeys, significant increases of 2',5'-oligoadenylate synthetase activity relative to IFN-alpha were maintained for 14 days in serum and the rHSA/IFNalpha2b showed more potent biological activity than IFN-alpha on a molar basis. Therefore, markedly improved in vivo biological activity of rHSA/IFNalpha2b could exhibit more potent antiviral activity than IFNalpha2b in future clinical trials. PMID- 17418550 TI - Assessment of the perinatal effects of maternal ingestion of Ipomoea carnea in rats. AB - It is believed that Ipomoea carnea toxicosis induces abnormal embryogenesis in livestock. Studies on rats treated with I. carnea aqueous fraction (AF) during gestation, revealed litters with decreased body weight, but the characteristic vacuolar lesions promoted by swainsonine, its main toxic principle, were observed only in young rats on postnatal day (PND) 7. However, these alterations could have resulted as consequence of swainsonine placental passage and/or damage or even ingestion of the contaminated milk by pups. Thus, this perinatal work was performed to verify the transplacental passage of swainsonine and its excretion into milk employing the cross-fostering (CF) procedure as a tool of study. Females were treated with AF or vehicle during gestation and after birth pups were fostered between treated and untreated dams. Pup body weight gain (BWG) and histopathology to observe vacuolar degeneration were performed on PND 3 and 7. In addition, swainsonine detection was performed in amniotic fluid and milk from rats treated with the AF during gestation or lactation. BWG was significantly lower only in pups from mothers treated with the plant and fostered to other treated mothers (AF-AF group of pups). The histopathology revealed that pups from treated mothers fostered to untreated ones showed the characteristic vacuolar lesions; however, the lesions from the AF-AF pups were more severe in both periods evaluated. Amniotic fluid and milk analysis revealed the presence of swainsonine excretion into these fluid compartments. Thus, the results from CF and the chemical analysis allowed concluding that swainsonine passes the placental barrier and affects fetal development and milk excretion participates in I. carnea perinatal toxicosis. PMID- 17418551 TI - Detecting the form of selection in the outer membrane protein C of Enterobacter aerogenes strains and Salmonella species. AB - The types of selective pressure operating on the outer membrane protein C (ompC) of Enterobacter aerogenes strains, the causative agent for nosocomial infections, and Salmonella sp., the hazardous pathogen are investigated using the maximum likelihood-based codon substitution models. Although the rate of amino acid replacement to the silent substitution (omega) across the entire codon sites of ompC of E. aerogenes (omega=0.3194) and Salmonella sp. (omega=0.2047) indicate that the gene is subjected to purifying selection (i.e. omega<1), approximately 3.7% of ompC codon sites in E. aerogenes (omega=21.52) are under the influence of positive Darwinian selection (i.e. omega>1). Such contrast in the intensity of selective pressures in both pathogens could be associated with the differential response to the adverse environmental changes. In E. aerogenes, majority of the positively selected sites are located in the hypervariable cell-surface-exposed domains whereas the trans-membrane domains are functionally highly constrained. PMID- 17418552 TI - Antibacterial and antifungal activity of cicerfuran and related 2-arylbenzofurans and stilbenes. AB - Cicerfuran, 2-(2-methoxy-4,5-methylenedioxyphenyl)-6-hydroxybenzofuran, is an antifungal phytoalexin previously isolated from the roots of chickpea, Cicer spp. The synthesis of cicerfuran, five 2-arylbenzofuran analogues and nine stilbene intermediates was reported recently. The antimicrobial activities of these compounds were evaluated against two species of bacteria, Bacillus subtilis and Pseudomonas syringae, and four species of filamentous fungi, Aspergillus niger, Botrytis cinerea, Cladosporium herbarum and Monilinia aucupariae. Stilbenes with a free hydroxyl group were active against both bacteria and fungi with MICs in the range 25-100microg/ml. Cicerfuran was the only 2-arylbenzofuran that showed antimicrobial activity with MICs as low as 25microg/ml. Some aspects of the structure-activity relationship are discussed. PMID- 17418553 TI - Resuscitation and quantification of stressed Escherichia coli K12 NCTC8797 in water samples. AB - The aim of this study was to investigate the impact on numbers of using different media for the enumeration of Escherichia coli subjected to stress, and to evaluate the use of different resuscitation methods on bacterial numbers. E. coli was subjected to heat stress by exposure to 55 degrees C for 1h or to light induced oxidative stress by exposure to artificial light for up to 8h in the presence of methylene blue. In both cases, the bacterial counts on selective media were below the limits of detection whereas on non-selective media colonies were still produced. After resuscitation in non-selective media, using a multi well MPN resuscitation method or resuscitation on membrane filters, the bacterial counts on selective media matched those on non-selective media. Heat and light stress can affect the ability of E. coli to grow on selective media essential for the enumeration as indicator bacteria. A resuscitation method is essential for the recovery of these stressed bacteria in order to avoid underestimation of indicator bacteria numbers in water. There was no difference in resuscitation efficiency using the membrane filter and multi-well MPN methods. This study emphasises the need to use a resuscitation method if the numbers of indicator bacteria in water samples are not to be underestimated. False-negative results in the analysis of drinking water or natural bathing waters could have profound health effects. PMID- 17418554 TI - Formoterol, montelukast, and budesonide in asthmatic children: effect on lung function and exhaled nitric oxide. AB - BACKGROUND: It has been proposed that asthma control may be achieved in part by minimizing airway inflammation. The simultaneous effects of inhaled steroids associated with long-acting beta-agonists and leukotriene antagonists on pulmonary function and airway inflammation are still largely unexplored in children with moderate persistent asthma. OBJECTIVES: The aim of this study was to investigate the effects of add-on therapy with long-acting beta-agonists and leukotriene antagonists on FEV1 and exhaled nitric oxide levels (FENO) in children. METHODS: Forty-eight steroid-naive atopic asthmatic children, 7-11 years of age, were randomly treated in four groups for two consecutive one-month periods, as follows: (1) first month: budesonide 200 microg twice daily; second month: budesonide 400 microg twice daily; (2) first month: budesonide 200 microg twice daily+formoterol 9 microg twice daily; second month: budesonide 200 microg twice daily+montelukast 5mg once daily; (3) first month: budesonide 200 microg twice daily+montelukast 5mg once daily; second month budesonide 200 microg+formoterol 9 microg twice daily; (4) first and second month: budesonide 400 microg twice daily. RESULTS: All treatments resulted in a significant increase in lung function and a decrease in FENO compared with values at baseline. Budesonide+montelukast in combination was the most effective treatment for reducing FENO levels. CONCLUSION: This study demonstrates that add-on therapy with montelukast plus low-dose budesonide is more effective than the addition of long-acting beta-agonists or doubling the dose of budesonide for controlling FENO in asthmatic children. PMID- 17418555 TI - Differential expression of genes involved with apoptosis, cell cycle, connective tissue proteins, fuel substrate utilization, inflammation and mitochondrial biogenesis in copper-deficient rat hearts: implication of a role for Nfkappab1. AB - We hypothesized that the increase in mitochondrial proliferation in hearts from copper-deficient rats is due to an increase in expression of the transcriptional factor peroxisomal-like proliferating related coactivator 1alpha (Ppargc1a), which regulates transcriptional activity for many of the genes that encode for mitochondrial proteins. In addition to several transcriptional factors implicated in mitochondrial biogenesis, we also looked at a number of genes involved in cell cycle regulation and fuel substrate utilization. Long-Evans rats were placed on either a copper-adequate (n=4) or copper-deficient (n=4) diet 3 days post weaning and remained on the diet for 5 weeks; their copper deficiency status was confirmed using previously established assays. Custom oligo arrays spotted with genes pertinent to mitochondrial biogenesis were hybridized with cRNA probes synthesized from the collected heart tissue. Chemiluminescent array images from both groups were analyzed for gene spot intensities and differential gene expression. Our results did not demonstrate any significant increase in Ppargc1a or its implicated targets, as we had predicted. However, consistent with previous data, an up-regulation of genes that encode for collagen type 3, fibronectin and elastin were found. Interestingly, there was also a significant increase in the expression of the transcriptional factor nuclear factor kappaB1 (Nfkappab1) in the copper-deficient treatment animals, compared to the control group, and this was confirmed by real time quantitative polymerase chain reaction. The results of this study merit the further investigation of the role of reactive oxidative species with regard to Nfkappab1 in the copper deficient rat heart. PMID- 17418556 TI - Obesity causes very low density lipoprotein clearance defects in low-density lipoprotein receptor-deficient mice. AB - We have reported that obese leptin-deficient mice (ob/ob) lacking the low-density lipoprotein receptor (LDLR(-/-)) develop severe hyperlipidemia and spontaneous atherosclerosis. In the present study, we show that obese leptin receptor deficient mice (db/db) lacking LDLR have a similar phenotype, even in the presence of elevated plasma leptin levels. We investigated the mechanism for the hyperlipidemia in obese LDLR(-/-) mice by comparing lipoprotein production and clearance rates in C57BL/6, ob/ob, LDLR(-/-) and ob/ob;LDLR(-/-) mice. Hepatic triglyceride production rates were equally increased ( approximately 1.4-fold, P<.05) in both LDLR(-/-) and ob/ob;LDLR(-/-) mice compared to C57BL/6 and ob/ob mice. LDL clearance was decreased ( approximately 1.3- fold, P<.01) to a similar extent in LDLR(-/-) and ob/ob;LDLR(-/-) mice compared to C57BL/6 and ob/ob controls. While VLDL clearance was delayed in LDLR(-/-) compared to C57BL/6 and ob/ob mice (2-fold, P<.001), this delay was exaggerated in ob/ob;LDLR(-/-) mice (3.8-fold, P<001). The VLDL clearance defects were due to decreased hepatic uptake compared to C57BL/6 (54% and 26% for LDLR(-/-) and ob/ob;LDLR(-/-), respectively, P<.001). When VLDL was collected from C57BL/6, ob/ob, LDLR(-/-), and ob/ob;LDLR(-/-) donors and injected into LDLR(-/-) recipient mice, counts remaining in the liver were 1.4-fold elevated in mice receiving LDLR(-/-) VLDL and 2-fold increased in mice receiving ob/ob;LDLR(-/-) VLDL compared to controls receiving C57BL/6 VLDL (P<.01). Thus, the increase in plasma lipoproteins in ob/ob;LDLR(-/-) mice is caused by delayed VLDL clearance. This appears to be due to defects in both the liver and the lipoproteins themselves in these obese mice. PMID- 17418557 TI - Extra virgin olive oil increases uncoupling protein 1 content in brown adipose tissue and enhances noradrenaline and adrenaline secretions in rats. AB - The effects of extra virgin olive oil (EV-olive oil) on triglyceride metabolism were investigated by measuring the degree of thermogenesis in interscapular brown adipose tissue (IBAT) and the rates of noradrenaline and adrenaline secretions in rats, both in vivo and in situ. In Experiment 1 (in vivo), rats were given an isoenergetic high-fat diet (30% fat diet) containing corn oil, refined olive oil, or EV-olive oil. After 28 days of feeding, the final body weight, weight gain, energy efficiency, perirenal adipose tissue and epididymal fat pad and plasma triglyceride concentrations were the lowest in the rats fed the EV-olive oil diet. The content of uncoupling protein 1 (UCP1) in IBAT and the rates of urinary noradrenaline and adrenaline excretions were the highest in the rats fed the EV olive oil diet. In Experiment 2 (in situ), the effects of the extract of the phenolic fraction from EV-olive oil and a compound having excellent characteristics as components of EV-olive oil, hydroxytyrosol, on noradrenaline and adrenaline secretions were evaluated. The intravenous administration of the extract of the phenolic fraction from EV-olive oil significantly increased plasma noradrenaline and adrenaline concentrations, whereas that of hydroxytyrosol had no effect. These results suggest that phenols except hydroxytyrosol in EV-olive oil enhance thermogenesis by increasing the UCP1 content in IBAT and enhancing noradrenaline and adrenaline secretions in rats. PMID- 17418558 TI - Red blood cell folate vitamer distribution in healthy subjects is determined by the methylenetetrahydrofolate reductase C677T polymorphism and by the total folate status. AB - BACKGROUND: Red blood cells (RBCs) represent a storage pool for folate. In contrast to plasma, RBC folate can appear in different biochemical isoforms. So far, only the methylenetetrahydrofolate reductase (MTHFR) 677 TT genotype has been identified as a determinant of RBC folate vitamer distribution. OBJECTIVE: The purpose of this study is to identify clinical and biochemical determinants of RBC folate vitamer distribution in healthy subjects. DESIGN: In an observational study, 109 subjects, aged 18 to 65 years, were studied. Red blood cell folate vitamers were analyzed using a liquid chromatography-tandem mass spectrometry method. Other variables recorded included vitamin B(2), B(6) and B(12) status, homocysteine, plasma and RBC S-adenosylhomocysteine and S-adenosylmethionine, renal function and the MTHFR C677T polymorphism. RESULTS: The MTHFR C677T genotype was the dominant determinant of nonmethylfolate accumulation. The median (range) nonmethylfolate/total folate ratio was 0.58% (0-12.2%) in the MTHFR CC group (n=55), 0.99% (0-14.3%) in the CT group (n=39) and 30.3% (5.7-73.3%) in the TT genotype group (n=15), P<.001. The 95th percentile for the nonmethylfolate/total folate ratio was 2.8% for the CC group, 9.1% for the CT group and 73.3% for the TT group. In the CC and CT genotype subjects, the T allele and total folate status were positively and independently correlated with nonmethylfolate accumulation, but the degree of nonmethylfolate accumulation in these subjects was usually minor compared with those with the TT genotype. None of the other studied variables was associated with nonmethylfolate accumulation. CONCLUSIONS: The MTHFR C677T genotype is the dominant determinant of nonmethylfolate accumulation in RBCs. In addition, high total folate status may contribute to minor to moderate nonmethylfolate accumulation in MTHFR CC and CT subjects. PMID- 17418559 TI - HUM5007, a novel combination of thermogenic compounds, and 3-acetyl-7-oxo dehydroepiandrosterone: each increases the resting metabolic rate of overweight adults. AB - This study tested the hypothesis that 3-acetyl-7-oxo-dehydroepiandrosterone alone (7-Keto) and in combination with calcium citrate, green tea extract, ascorbic acid, chromium nicotinate and cholecalciferol (HUM5007) will increase the resting metabolic rate (RMR) of overweight subjects maintained on a calorie-restricted diet. In this randomized, double-blind, placebo-controlled, crossover trial, overweight adults on a calorie-restricted diet were randomized to three 7-day treatment periods with 7-Keto, HUM5007 or placebo. Resting metabolic rate was measured by indirect calorimetry at the beginning and end of each treatment period with a 7-day washout between testing periods. Of 45 subjects enrolled, 40 completed the study (30 women, 10 men; mean age, 38.5 years; mean mass index, 32.0 kg/m(2)). During the placebo treatment, RMR decreased by 3.9% (75+/-111 kcal/day; mean+/-S.D.); however, RMR increased significantly by 1.4% (21+/-115 kcal/day) and 3.4% (59+/-118 kcal/day) during the 7-Keto and HUM5007 treatment periods, respectively (each compared to placebo, P=.001). No significant differences were found between the treatment periods with respect to compliance or adverse events. In this study, the administration of HUM5007 or 7-Keto reversed the decrease in RMR normally associated with dieting. HUM5007 and 7-Keto increased RMR above basal levels and may benefit obese individuals with impaired energy expenditure. HUM5007 and 7-Keto were generally well tolerated and no serious adverse events were reported. PMID- 17418560 TI - Dietary factors and growth and metabolism in experimental tumors. AB - Development of a diet that provides adequate nutrition and effective cancer prevention is an important goal in nutrition and cancer research. A confounding aspect of dietary control of tumor growth is the fact that some nutrients may up regulate tumor growth, whereas other nutrients and nonnutrients down-regulate growth. Both up- and down-regulators may be present in the same foodstuff. Identification of these substances, determination of their mechanisms of action and potencies, as well as the interactions among the different mechanisms are topics of ongoing research. In this review, we describe results obtained in vivo or during perfusion in situ using solid tissue-isolated rodent tumors and human cancer xenografts in nude rats. Linoleic acid (LA), an essential n-6 polyunsaturated fatty acid (PUFA), was identified as an agent in dietary fat that is responsible for an up-regulation of tumor growth in vivo. Tumor LA uptake, mediated by high intratumor cAMP, stimulated formation of the mitogen, 13 hydroxyoctadecadienoic acid (13-HODE) and also increased ERK1/2 phosphorylation, [(3)H]thymidine incorporation and growth. A mechanism for control of this growth promoting pathway was revealed during studies of the effects of dietary nutrients and nonnutrients known to inhibit tumor growth. These included four groups of lipophilic agents: n-3 fatty acids, melatonin, conjugated LA isomers and trans fatty acids. Each of these agents activated an inhibitory G protein-coupled receptor-mediated pathway that specifically suppressed tumor uptake of saturated, monounsaturated and n-6 PUFAs, thereby inhibiting an early step in the LA dependent growth-promoting pathway. PMID- 17418561 TI - Piperine protects cisplatin-induced apoptosis via heme oxygenase-1 induction in auditory cells. AB - Piperine is a major component of black pepper, Piper nigrum Linn, used widely in traditional medicine. In this study, we examined whether piperine could protect House Ear Institute-Organ of Corti 1 (HEI-OC1) cells against cisplatin-induced apoptosis through the induction of heme oxygenase (HO)-1 expression. Piperine (10 100 microM) induced the expression of HO-1 in dose- and time-dependent manners. Piperine also induced antioxidant response element-luciferase and translocated nuclear factor-E2-related factor-2 (Nrf2) to nucleus. Piperine activated the c Jun N-terminal kinase (JNK), extracellular signal-regulated kinase and p38 mitogen-activated protein kinase (MAPK) pathways, and the JNK pathway played an important role in piperine-induced HO-1 expression. Piperine protected the cells against cisplatin-induced apoptosis. The protective effect of piperine was abrogated by zinc protoporphyrin IX, an HO inhibitor, and antisense oligodeoxynucleotides against HO-1 gene. These results demonstrate that the expression of HO-1 by piperine is mediated by both JNK pathway and Nrf2, and the expression inhibits cisplatin-induced apoptosis in HEI-OC1 cells. PMID- 17418562 TI - LCA of integrated MSW management systems: case study of the Bologna District. AB - LCA as a decision-supporting tool in planning integrated municipal solid waste management is not, as yet, widely used in Italy. This paper presents a study concerning the application of the LCA methodology to support the development of the new waste management plan for the Bologna District. The main goal of the study was to show decision-makers at the political level the benefits obtainable with the use of LCA, in terms of the identification and quantification of the potential environmental impacts of different waste management strategies. The integrated waste management system of the Bologna District includes waste collection and transport, sorting, recycling, composting, incineration and landfilling. Three scenarios, referring to 2006 and assuming the presence of 950,000 inhabitants and the production of approximately 566,000 t of waste in the district, have been compared. A detailed model has been developed in order to capture effects related to the waste fraction from separated collection and to the different waste treatments. The discussion of the results has focussed in particular on the greenhouse effect and the acidification potential. On the basis of the results obtained, the analysis of an additional scenario characterised by a further increase in separated collection has been put forward. PMID- 17418563 TI - A protein microarray prepared with phage-displayed antibody clones. AB - Using a large phage antibody library, a protein microarray spotted directly with phage-displayed antibody clones was created to discriminate between recognition profiles of samples from healthy donors and leukemia patients. The protocol for preparing antibody-displaying phage chips was presented. Some conditions such as substrates and blocking buffers were compared and optimized. The major improvements of this microarray are higher throughput and lower cost compared to previous antibody chips. Due to its convenience and sensitivity, it can be extensively used for rapid and high throughput detection of protein profiles of experimental and clinical samples. PMID- 17418564 TI - Evaluating the quality of communities made compost manure in South Africa: a case study of content and sources of metals in compost manure from Thulamela Municipality, Limpopo province. AB - The main aim of this research was to assess the quality of the compost manure of a community project which is part of Government's initiative for job creation. This was done by determining the sources, presence and concentrations of metals in the composting materials and compost manure. Generally, there are many communities involved in making compost manure with no assessment of the quality of the compost. This is because no quality assurance guidelines are in place. The main source of heavy metals in the composting materials was found to be sawdust while cabbage was the main source of the essential nutrient elements such as magnesium calcium and potassium. The level of heavy metals in all the compost materials were found in the following order: Fe>Mn>Cr>Ni>Zn>Cu>Co>Cd. The level of essential elements was in the following order: Ca>Mg>K>S>Na>Li>Si>P. The level of heavy metals in the compost materials reflected the level in the final compost manure. The concentration of metals in the compost materials and compost manure was below the international standards of compost manure. Therefore, compost manure can be said to be of good quality. However, sawdust was found to be not a good compost material as it was the source of main heavy metals. PMID- 17418565 TI - Heterocyclic nitrogenous pollutants in the environment and their treatment options--an overview. AB - Heterocyclic nitrogenous bases are one of the most important class of compounds containing N as a heteroatom, like pyridine and its derivatives. These compounds are of immense concern from point view of environment since they are known for their toxic and carcinogenic properties, lethal effect on natural biogenic environment and severe odour potential. There is a need to control these compounds from getting discharged into the environment. This paper addresses the different natural/anthropogenic sources which generate these pollutants, their toxicity profile, different physico-chemical treatment methods and especially focuses on biological methods of treatment and combination of these for the efficient removal to achieve a treated effluent quality fit for disposal without causing any damage to the environment. PMID- 17418566 TI - Enzymatic synthesis of arbutin undecylenic acid ester and its inhibitory effect on melanin synthesis. AB - Transesterification of arbutin and undecylenic acid vinyl ester was catalyzed by alkaline protease, Bioprase, in dimethylformamide to get arbutin derivative having undecylenic acid at 6-position of glucose moiety, 6-O-undecylenoyl p hydroxyphenyl beta-D-glucopyranoside. The reaction rate increased with increase of arbutin concentration, and when its concentration was 0.9 M, the conversion rate was more than 90% under addition of 2 M undecylenic acid vinyl ester. The obtained arbutin ester significantly suppressed melanin production in murine B16 melanoma cells. PMID- 17418567 TI - Evaluation of N-(phenylmethyl)-4-[5-(phenylmethyl)-4,5,6,7-tetrahydro-1H imidazo[4,5-c]pyridin-4-yl]benzamide inhibitors of Mycobacterium tuberculosis growth. AB - The biological evaluation of imidazopiperidines as FAS II inhibitors of Mycobacterium tuberculosis growth has been carried out with a view to assessment of potential as lead compounds for the development of a new TB drug. A summary of the hit evaluation and current challenges is described herein. PMID- 17418568 TI - 1,3-disubstituted 4-aminopiperidines as useful tools in the optimization of the 2 aminobenzo[a]quinolizine dipeptidyl peptidase IV inhibitors. AB - In a search for novel DPP-IV inhibitors, 2-aminobenzo[a]quinolizines were identified as submicromolar HTS hits. Due to the difficult synthetic access to this compound class, 1,3-disubstituted 4-aminopiperidines were used as model compounds for optimization. The developed synthetic methodology and the SAR could be transferred to the 2-aminobenzo[a]quinolizine series, leading to highly active DPP-IV inhibitors. PMID- 17418569 TI - Ligand based virtual screening and biological evaluation of inhibitors of chorismate mutase (Rv1885c) from Mycobacterium tuberculosis H37Rv. AB - We have identified new lead candidates that possess inhibitory activity against Mycobacterium tuberculosis H37Rv chorismate mutase by a ligand-based virtual screening optimized for lead evaluation in combination with in vitro enzymatic assay. The initial virtual screening using a ligand-based pharmacophore model identified 95 compounds from an in-house small molecule database of 15,452 compounds. The obtained hits were further evaluated by molecular docking and 15 compounds were short listed based on docking scores and the other scoring functions and subjected to biological assay. Chorismate mutase activity assays identified four compounds as inhibitors of M. tuberculosis chorismate mutase (MtCM) with low K(i) values. The structural models for these ligands in the chorismate mutase binding site will facilitate medicinal chemistry efforts for lead optimization against this protein. PMID- 17418570 TI - CC chemokine receptor-3 (CCR3) antagonists: improving the selectivity of DPC168 by reducing central ring lipophilicity. AB - DPC168, a benzylpiperidine-substituted aryl urea CCR3 antagonist evaluated in clinical trials, was a relatively potent inhibitor of the 2D6 isoform of cytochrome P-450 (CYP2D6). Replacement of the cyclohexyl central ring with saturated heterocycles provided potent CCR3 antagonists with improved selectivity against CYP2D6. The favorable preclinical profile of DPC168 was maintained in an acetylpiperidine derivative, BMS-570520. PMID- 17418571 TI - Design, synthesis, and evaluation of tetrahydroquinoline and pyrrolidine sulfonamide carbamates as gamma-secretase inhibitors. AB - Gamma-secretase is a key enzyme involved in the production of beta-amyloid peptides which are believed to play a critical role in the onset and progression of Alzheimer's disease (AD). As such, inhibition of gamma-secretase has been an attractive approach to AD therapy. In this paper, the design, synthesis, and evaluation of tetrahydroquinoline and pyrrolidine sulfonamide carbamates as gamma secretase inhibitors are described. PMID- 17418572 TI - Metabolic activation of indole-containing prostaglandin D2 receptor 1 antagonists: impacts of glutathione trapping and glucuronide conjugation on covalent binding. AB - Some DP1 receptor antagonists from an indole-containing series were shown to cause in vitro covalent binding to protein in rat and human liver microsomes. Glutathione trapping experiments along with in vitro labeling assays confirmed that the presence of a strong electron withdrawing group was necessary to abrogate in vitro covalent binding, leading to the discovery of MK-0524. Hepatocyte incubations and in vivo studies showed that acyl-glucuronide formation did not translate into covalent binding. PMID- 17418573 TI - Hypokalaemic periodic paralysis due to the CACNA1S R1239H mutation in a large African family. AB - Hypokalaemic periodic paralysis (HypoKPP) is a skeletal muscle channelopathy caused by mutations in calcium (CACNA1S) and sodium (SCN4A) channel subunits. A small number of causative mutations have been found in European and Asian patients, but not in African patients yet. We have identified a large Beninese family in which HypoKPP segregated over five generations and was caused by the CACNA1S R1239H mutation. We report on the clinical and histopathological spectrum of the disorder in this family. A later age at onset (15.8+/-8.8years), and particular triggering factors due to specific African life conditions seem to be characteristic of our observation. PMID- 17418574 TI - Phenotypic patterns of desminopathy associated with three novel mutations in the desmin gene. AB - Desminopathy represents a subgroup of myofibrillar myopathies caused by mutations in the desmin gene. Three novel disease-associated mutations in the desmin gene were identified in unrelated Spanish families affected by cardioskeletal myopathy. A selective pattern of muscle involvement, which differed from that observed in myofibrillar myopathy resulting from mutations in the myotilin gene, was observed in each of the three families with novel mutations and each of three desminopathy patients with known desmin mutations. Prominent joint retractions at the ankles and characteristic nasal speech were observed early in the course of illness. These findings suggest that muscle imaging in combination with routine clinical and pathological examination may be helpful in distinguishing desminopathy from other forms of myofibrillar myopathy and ordering appropriate molecular investigations. PMID- 17418575 TI - Exemestane after non-steroidal aromatase inhibitors for post-menopausal women with advanced breast cancer. AB - A retrospective analysis was performed on 31 consecutive locally advanced or metastatic breast cancer patients who commenced exemestane 25mg/d orally following previous treatment with Tamoxifen and a non-steroidal third-generation aromatase inhibitor (AI). Patients were seen 3 monthly until clinical or radiological disease progression. Median age was 64 years (range 34-90 yrs). The average number of recurrences before starting exemestane was three (range 1-6). There were two complete responses (CR), four partial responses (PR), 12 with stable disease (SD) and 12 with progressive disease (PD). Objective response rate (CR+PR) was 19.4% and overall clinical benefit (CR+PR+SD >or= 24 weeks) was 54.8%. The median durations of objective response and overall clinical benefit were 18 and 14 months, respectively. This data support the anti-tumour activity of exemestane 25mg daily in patients with locally advanced and/or metastatic breast cancer who have been previously exposed to non-steroidal AIs and Tamoxifen. PMID- 17418576 TI - Favorable outcome in patients with breast cancer in the presence of pathological response after neoadjuvant endocrine therapy. AB - Neoadjuvant endocrine therapy (NAET) can expand the number of breast cancer patients who can be treated with breast-conserving surgery and can predict benefit from adjuvant endocrine therapy. Because no validated surrogate markers for long-term outcome have been established, we conducted prospective trials to evaluate pathological response and Ki-67 index following treatment with tamoxifen or anastrozole. The study population included postmenopausal women with operable breast tumors that were both estrogen and progesterone receptor-positive and larger than 3 cm. Response was classified as pathological response (minimal response or better) and non-response. Non-responding (25.5%, vs. response 85.9%, p=0.002), axillary node-positive (58.4% vs. node negative 100%, p=0.045), and high pretreatment Ki-67 index (41.4% vs. low Ki-67 87.1%, p=0.03) patients were significantly associated with poor 5-year relapse-free survival. Multivariate analysis of relapse-free survival indicated that pathological response was independent. Therefore, pathological response may be a favorable prognostic factor after NAET. PMID- 17418577 TI - Going for baroque at the Escherichia coli K1 cell surface. AB - Phase variation is usually thought of as the stochastic switching between alternatively expressed ('on') and unexpressed ('off') phenotypic states. However, coupling synthesis of a monotonous homopolysaccharide to a mechanism of random but incomplete chemical modification produces almost infinite structural variation. Potentially limitless variability implies that evolution can produce highly ornate or extravagant flourishes reminiscent of the baroque style. Here, we describe an analysis of capsular polysialic acid form variation in Escherichia coli K1, demonstrating that the large number of variant structures is controlled by a single contingency locus. The mechanism for generating maximum structural diversity from maximal genetic parsimony is conferred by a simple translational switch carried on a K1-specific prophage. PMID- 17418578 TI - Preparation, biological activity and endogenous occurrence of N6 benzyladenosines. AB - Cytokinin activity of forty-eight 6-benzyladenosine derivatives at both the receptor and cellular levels as well as their anticancer properties were compared in various in vitro assays. The compounds were prepared by the condensation of 6 chloropurine riboside with corresponding substituted benzylamines and characterized by standard collection of physico-chemical methods. The majority of synthesized derivatives exhibited high activity in all three of the cytokinin bioassays used (tobacco callus, wheat leaf senescence and Amaranthus bioassay). The highest activities were observed in the senescence bioassay. For several of the compounds tested, significant differences in activity were found between the bioassays used, indicating that diverse recognition systems may operate. This suggests that it may be possible to modulate particular cytokinin-dependent processes with specific compounds. In contrast to their high activity in bioassays, the tested compounds were recognized with only very low sensitivity in both Arabidopsis thaliana AHK3 and AHK4 receptor assays. The prepared derivatives were also investigated for their antiproliferative properties on cancer and normal cell lines. Several of them showed very strong cytotoxic activity against various cancer cell lines. On the other hand, they were not cytotoxic for normal murine fibroblast (NIH/3T3) cell line. This anticancer activity of cytokinin ribosides may be important, given that several of them occur as endogenous compounds in different organisms. PMID- 17418579 TI - QSAR study on permeability of hydrophobic compounds with artificial membranes. AB - We previously reported a classical quantitative structure-activity relationship (QSAR) equation for permeability coefficients (P(app-pampa)) by parallel artificial membrane permeation assay (PAMPA) of structurally diverse compounds with simple physicochemical parameters, hydrophobicity at a particular pH (logP(oct) and |pK(a)-pH|), hydrogen-accepting ability (SA(HA)), and hydrogen donating ability (SA(HD)); however, desipramine, imipramine, and testosterone, which have high logP(oct) values, were excluded from the derived QSAR equation because their measured P(app-pampa) values were lower than calculated. In this study, for further investigation of PAMPA permeability of hydrophobic compounds, we experimentally measured the P(app-pampa) of more compounds with high hydrophobicity, including several pesticides, and compared the measured P(app pampa) values with those calculated from the QSAR equation. As a result, compounds having a calculated logP(app-pampa)>-4.5 showed lower measured logP(app pampa) than calculated because of the barrier of the unstirred water layer and the membrane retention of hydrophobic compounds. The bilinear QSAR model explained the PAMPA permeability of the whole dataset of compounds, whether hydrophilic or hydrophobic, with the same parameters as the equation in the previous study. In addition, PAMPA permeability coefficients correlated well with Caco-2 cell permeability coefficients. Since Caco-2 cell permeability is effective for the evaluation of human oral absorption of compounds, the proposed bilinear model for PAMPA permeability could be useful for not only effective screening for several drug candidates but also the risk assessment of chemicals and agrochemicals absorbed by humans. PMID- 17418580 TI - Application of descriptors based on Lipinski's rules in the QSPR study of aqueous solubilities. AB - We complement new physically interpretable descriptors inspired by the Lipinski's rules of drug bioavailability with others obtained from the Dragon 3.0 software, in order to find the best QSPR relationship for aqueous solubilities of 100 structurally heterogeneous organic, drug-like compounds. The simultaneous linear regression analyses of 1367 variables lead to a six-parameter model containing two of the new proposed descriptors and which also possess good predictive ability given by R=0.8798 and cross-validated R(1-10%-o)=0.8199. We further validate the model found with an external test set composed of 48 compounds. PMID- 17418581 TI - The structure-activity relationship of the series of non-peptide small antagonists for p56lck SH2 domain. AB - The antagonists for the SH2 domain are regarded as novel therapeutic candidates for cancer, autoimmune disease, and chronic inflammatory disease. Previously, we identified rosmarinic acid (alpha-o-caffeoyl-3,4-dihydroxyphenyl-lactic acid; RosA) from Prunella vulgaris as an antagonist for the p56lck SH2 domain by screening natural products. RosA not containing phosphotyrosine surrogate had a considerable inhibitory activity for T-cell antigen receptor (TCR)-induced interleukin (IL)-2 expression, and subsequent T-cell proliferation in vitro cell assay. To investigate the structure-activity relationship of RosA and to identify a novel p56lck SH2 antagonist with more potent in vitro T-cell inhibitory activity, we synthesized several analogs of RosA by using rational design. All synthesized compounds were tested in vitro binding activity for the SH2 domain and in vitro T-cell inhibitory activity. All four hydroxyl groups of RosA were essential for binding with the p56lck SH2 domain and T-cell inhibitory activity. Unexpectedly, conformationally less constrained analogs 4 and 9 showed a more potent binding affinity for the SH2 domain than that of RosA, and chirality of the analog did not play an important role in protein binding. We successfully identified several RosA analogs with a more potent T-cell inhibitory activity than that of RosA. Overall results revealed important structural requirements of the p56lck SH2 antagonists for in vitro T-cell inhibitory activity and in vitro protein binding activity. PMID- 17418582 TI - Synthesis and topoisomerase poisoning activity of A-ring and E-ring substituted luotonin A derivatives. AB - A series of A-ring and E-ring analogues of the natural product luotonin A, a known topoisomerase I poison, was evaluated for growth inhibition in human carcinoma and leukemia cell lines. Rational design of structures was based on analogues of the related alkaloid camptothecin, which has been demonstrated to exert cytotoxic effects by the same mechanism of action. When compared to luotonin A, several compounds exhibited an improved topoisomerase I-dependent growth inhibition of a human leukemia cell line. PMID- 17418583 TI - Synthesis, structure, and structure-activity relationship analysis of enamines as potential antibacterials. AB - Twenty-four enamines were synthesized and reported for the first time. Their chemical structures were confirmed by means of 1H NMR, ESI mass spectra, and elemental analyses, and four of them were determined by single crystal X-ray diffraction analysis. All of the compounds were assayed for antibacterial (Bacillus subtilis ATCC 6633, Escherichia coli ATCC 35218, Pseudomonas fluorescens ATCC 13525, and Staphylococcus aureus ATCC 6538) and antifungal (Aspergillus niger ATCC 16404, Candida albicans ATCC 10231, and Trichophyton rubrum ATCC 10218) activities by MTT method. Compounds (E)-ethyl 3-(4 hydroxyphenylamino)-2-(4-methoxyphenyl)acrylate (9b), (E)-ethyl 3-(3,5 difluorophenylamino)-2-(4-chlorophenyl)acrylate (11b), (E)-ethyl 3-(3,5 dichlorophenylamino)-2-(4-chlorophenyl)acrylate (12b), and (E)-ethyl 3-(4 methylphenylamino)-2-(4-chlorophenyl)acrylate (15b) showed considerable antibacterial activities against S. aureus ATCC 6538 with MICs of 3.8, 1.9, 1.1, and 0.9 microg/mL, respectively. Structure-activity relationship (SAR) analysis disclosed, generally, an E-isomer exhibited higher antibacterial activity than the corresponding Z-isomer. An electron-withdrawing group on A-ring led to some decrease in activity, while on B-ring, a similar substitution provided higher activity. PMID- 17418584 TI - Differential expression of molecular motors in the motor cortex of sporadic ALS. AB - The molecular mechanisms underlying the selective neurodegeneration of motor neurons in amyotrophic lateral sclerosis (ALS) are inadequately understood. Recent breakthroughs have implicated impaired axonal transport, mediated by molecular motors, as a key element for disease onset and progression. The current work identifies the expression of 15 kinesin-like motors in healthy human motor cortex, including three novel isoforms. Our comprehensive quantitative mRNA analysis in control and sporadic ALS (SALS) motor cortex specimens detects SALS specific down-regulation of KIF1Bbeta and novel KIF3Abeta, two isoforms we show to be enriched in the brain, and also of SOD1, a key enzyme linked to familial ALS. This is accompanied by a marked reduction of KIF3Abeta protein levels. In the motor cortex KIF3Abeta localizes in cholinergic neurons, including upper motor neurons. No mutations causing splicing defects or altering protein-coding sequences were identified in the genes of the three proteins. The present study implicates two motor proteins as possible candidates in SALS pathology. PMID- 17418585 TI - Soluble expanded PABPN1 promotes cell death in oculopharyngeal muscular dystrophy. AB - Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant disease caused by the expansion of a polyalanine repeat (GCG)(8-13) in exon 1 of the PABPN1 gene. Skeletal muscle fibers nuclei from OPMD patients contain insoluble polyalanine expanded PABPN1 (expPABPN1) nuclear aggregates that sequester different cellular components. Whether these aggregates are pathogenic, or the consequence of a molecular defense mechanism, remains controversial in the field of neurodegenerative disorders and OPMD. Our cellular model shows that interfering with the formation of expPABPN1-induced large nuclear aggregates increases the availability of nuclear expPABPN1 and significantly exacerbates cell death. Live microscopy reveals that cells harboring an increased amount of the soluble forms of expPABPN1 are significantly more prone to toxicity than those with nuclear aggregates. This is the first report directly indicating that nuclear aggregation in OPMD may reflect an active process by which cells sequester and inactivate the soluble toxic form of expPABPN1. PMID- 17418586 TI - Mortality and risk factors for surgical lung biopsy in patients with idiopathic interstitial pneumonia. AB - BACKGROUND: The overall safety of surgical lung biopsy in patients with idiopathic interstitial pneumonia (IIP) remains controversial. This study was performed to investigate the mortality and complication rate and identify the risk factors for surgical lung biopsy in patients with IIP. METHODS: A total of 200 patients with IIP who underwent surgical lung biopsy at the Asan Medical Center, Korea, from April 1990 to August 2003, were enrolled. Complications and mortality were analyzed retrospectively. RESULTS: (1) The mortality rate 30 days after the surgical lung biopsy was 4.3%, which was significantly higher than the control group. Biopsy performed at the time of acute exacerbation (AE) resulted in higher 30-day mortality (28.6%) compared to non-AE (3.0%; p<0.05). AE was followed by biopsy itself in three cases. (2) Univariate analysis indicated that lower FVC, lower DL(CO), and presence of AE were significant risk factors for 30 day mortality (p<0.05). However, multivariate analysis revealed that only AE (OR: 11.334, 95% CI: 1.727-74.365, p=0.011) was an independent risk factor. (3) The patients with low DL(CO) (<50% predicted) had higher mortality and complication rate than high DL(CO) group. CONCLUSION: Our data suggested that the presence of acute exacerbation at the time of biopsy and lower DL(CO) were predictors of higher mortality after the surgical lung biopsy. PMID- 17418587 TI - Genetic testing in patients with aortic aneurysms/dissections: a novel genotype/phenotype correlation? AB - OBJECTIVE: Mutations in the genes encoding fibrillin-1 (FBN1) and transforming growth factor beta receptor type II (TGFBR2) are known causes of Marfan syndrome (MFS) and related disorders. However, a sound correlation between the genotype and the cardiovascular phenotype has not yet been established. The objective of the present study was to identify novel mutations in FBN1 and TGFBR2 and to assess whether the type of mutation is linked to a particular clinical subtype of the cardiovascular condition. METHODS: The clinical records of 36 patients referred to us for molecular genetic diagnosis were reviewed to assess the course and severity of the vascular deterioration. A semiautomatic protocol was established enabling a rapid and cost-effective screening of the genes FBN1 and TGFBR2 by direct sequencing of all coding exons and flanking intronic regions. RESULTS: Novel mutations in FBN1 and TGFBR2 were detected in 12 and 2 patients, respectively. Four individuals carried a recurrent mutation in FBN1. Throughout the study cohort, the incidence of aortic dissections per se did not depend on the type of mutation. However, we found that mutations affecting the calcium binding epidermal growth factor-like domain were more frequently associated with a dissection of distal parts of the aorta than mutations that lead to a premature termination codon (chi(1)(2): p=0.013), suggesting that the spatio-temporal pattern of vascular deterioration may vary with the type of mutation. CONCLUSIONS: Detecting a mutation in the genes FBN1 and TGFBR2 proves the genetic origin of vascular findings and allows the identification of family members at risk who should undergo preventive checkups. Routine genetic testing of patients with suspected MFS or thoracic aortic aneurysms/dissections could provide further insight into genotype/phenotype correlations related to aortic dissection. PMID- 17418588 TI - Contribution of modelling chemotherapy-induced hematological toxicity for clinical practice. AB - Anticancer chemotherapies are responsible for numerous adverse events. Among these, hematological toxicity is one of the main causes for ending treatment. These toxicities decrease production of red blood cells (anemia), production of white blood cells (neutropenia or granulocytopenia), and production of platelets (thrombocytopenia), which may be life-threatening to the patient. Preventing such discontinuation would be valuable for treating patients more effectively. In order to achieve this goal, numerous mathematical and physiological or semiphysiological models have been developed. The complexity of models has increased over the years, from empiric E(max) models to mechanistic models including physiological mechanisms such as feedback control. This review discusses several approaches of modelling hematological toxicities illustrated with some examples: pharmacodynamic models for the hematological toxicity of 5 fluorouracil, epirubicin, melphalan, paclitaxel, topotecan, and indisulam. PMID- 17418589 TI - Formation of (bn-1 + H2O) ions by collisional activation of MALDI-formed peptide [M + H]+ ions in a QqTOF mass spectrometer. AB - Collisional activation of [M + H](+) parent ions from peptides of n amino acid residues may yield a rearrangement that involves loss of the C-terminal amino acid residue to produce (b(n-1) + H(2)O) daughters. We have studied this reaction by a retrospective examination of the m/z spectra of two collections of data. The first set comprised 398 peptides from coat protein digests of a number of plant viruses by various enzymes, where conditions in the tryptic digests were chosen so as to produce many missed cleavages. In this case, a large effect was observed 323 (b(n-1) + H(2)O) daughter ions (approximately 81%), including 185 (approximately 46%) "strong" decays with ratios (b(n-1) + H(2)O)/(b(n-1)) > 1. The second set comprised 1200 peptides, all from tryptic digests, which were carried out under more stringent conditions, resulting in relatively few missed cleavages. Even here, 190 (b(n-1) + H(2)O) ions (approximately 16%) were observed, including 87 (> 7%) "strong" decays, so the effect is still appreciable. The results suggest that the tendency for (b(n-1) + H(2)O) ion formation is promoted by the protonated side chain of a non-C-terminal basic amino acid residue, in the order arginine >> lysine > or = histidine, and that its (non-C-terminal) position is not critical. The results can be interpreted by a mechanism in which hydrogen bonding between the protonated side chain and the (n - 1) carbonyl oxygen facilitates loss of the C-terminal amino acid residue to give a product ion having a carboxyl group at the new C-terminus. PMID- 17418590 TI - Production of an anti-mouse MHC class II monoclonal antibody with biological activity in transgenic tobacco. AB - To produce a monoclonal antibody specific to a mouse major histocompatibility complex (MHC) class II protein, we synthesized the complementary DNAs for the heavy and light chains of a monoclonal antibody by PCR amplification. These cDNAs were then introduced separately into tobacco plant cells. After performing Northern blot analysis to confirm the expression of each of the chain genes in the transformed plants, we constructed transgenic plants expressing both the heavy and light chains by sexual crossing. The expression of the heavy and light chain genes in the sexually crossed plant was confirmed by Northern and Western blot analyses, respectively. Fluorocytometric analysis showed that the plant derived antibodies, which we purified using a protein G affinity column, bound specifically to target cells that expressed the cognate MHC class II molecules on their cell surfaces. The results of this study demonstrate that a monoclonal antibody against mouse MHC class II proteins can be expressed in transgenic plants. They also show the specific binding activity of plant-derived antibodies to cognate antigens. PMID- 17418591 TI - Neural mechanisms of auditory awareness underlying verbal transformations. AB - Prolonged listening to a repeated word without a pause produces a series of illusory transitions of the physically unchanging word, which is called verbal transformation. Verbal transformations provide a rare opportunity to examine how auditory percepts are formed in the brain. We found that verbal forms are affected by phonetic reorganization of a word, rather than by auditory adaptation and lexical distortion of it. We identified brain activity leading to individual differences between perceptual transitions and tone detection. An event-related fMRI analysis revealed that the left inferior frontal cortex (IFC), anterior cingulate cortex (ACC), and the left prefrontal cortex were activated when perceptual transitions from one verbal form to another occurred, but not when tone pips were detected. The number of perceptual transitions showed positive and negative correlations with signal intensity in the left IFC and the left ACC, respectively. The results suggest that active generation of verbal forms is linked with articulatory gestures for speech production and that the frequency of perceptual transitions is determined by a balance of the activations between the two brain regions. Structural equation modeling demonstrated that individual differences in the number of perceptual transitions rely on negative feedback from the ACC to the IFC via the posterior insula. These findings suggest that distributed frontal areas are involved in auditory awareness underlying verbal transformations. PMID- 17418592 TI - The use of standardized infinity reference in EEG coherency studies. AB - The study of large scale interactions in the brain from EEG signals is a promising method for the identification of functional networks. However, the validity of a large scale parameter is limited by two factors: the use of a non neutral reference and the artifactual self-interactions between the measured EEG signals introduced by volume conduction. In this paper, we propose an approach to study large scale EEG coherency in which these factors are eliminated. Artifactual self-interaction by volume conduction is eliminated by using the imaginary part of the complex coherency as a measure of interaction and the Reference Electrode Standardization Technique (REST) is used for the approximate standardization of the reference of scalp EEG recordings to a point at infinity that, being far from all possible neural sources, acts like a neutral virtual reference. The application of our approach to simulated and real EEG data shows that the detection of interaction, as opposed to artifacts due to reference and volume conduction, is a goal that can be achieved from the study of a large scale parameter. PMID- 17418593 TI - Men with epilepsy--the lost tribe? Results of a nationwide survey of men with epilepsy living in the UK. AB - The incidence of epilepsy is the same in men and women yet more time and energy has been expended on "women's issues" in recent times. In 2004, Epilepsy Action (The British Epilepsy Association) conducted a nationwide survey of men with epilepsy (MWE), who had contacted their helpline in the previous year. The men were comfortable discussing their epilepsy and confident in asking for information. They felt keenly the lack of driving privileges and the type of work available to them. A majority felt the condition adversely affected their self esteem and quality of life. Most men expressed satisfaction with the quality of care and information they got from their general practitioners and their neurologist. PMID- 17418594 TI - Transforming growth factor beta-1 enhances cytotoxic effect of doxorubicin in human lung adenocarcinoma cells of A549 line. AB - Transforming growth factor beta-1 (TGFbeta-1) is a regulator of cell proliferation, differentiation and apoptosis. Doxorubicin (adriamycin), an anthracycline drug causing double-strand DNA breaks, is widely used in anticancer chemotherapy. Here we demonstrated that TGFbeta-1 enhanced cytotoxic (proapoptotic) action of doxorubicin towards cultured human lung carcinoma A549 cells. Western-blot analysis and immunocytochemistry were used to show that doxorubicin induced PARP degradation in A549 cells, and TGFbeta-1 enhanced that action of the drug. The obtained results suggest a possibility of biomodulating effect of TGFbeta-1 on tumor cell treatment with doxorubicin. PMID- 17418595 TI - Cisplatin cytotoxicity is enhanced with zoledronic acid in A549 lung cancer cell line: preliminary results of an in vitro study. AB - We tested whether zoledronic acid, a biphosphonate with proposed apoptotic activity, augmented the cytotoxicity of cisplatin and/or gemcitabine in A549 lung cancer cell line. This cell line was subjected to different concentrations of the above chemotherapeutic agents and zoledronic acid. Cytotoxicity was assessed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide) assay. Particularly, zoledronic acid in 100 micromolar (microM) concentration augmented the cytotoxicity by cisplatin 1microg/ml from 25% to 70% (Z=3.22, P=0.0072). A significant portion of cells underwent apoptosis with or without zoledronic acid, but more so with the combination treatment as assessed by an Annexin V-FITC apoptosis detection kit. However, 100microM zoledronic acid showed 50% cytotoxicity on its own, but failed to improve cytotoxicity by Gemcitabine. Thus, we show for the first time in a lung cancer cell line that zoledronic acid bears cytotoxic potential on its own and in conjunction with cisplatin. The clinical potential of this finding should be further studied. PMID- 17418596 TI - A systematic review of the neurobehavioural and physiological effects of shiftwork systems. AB - Shiftwork is a common experience for many workers. There are a wide range of shift systems in use, with a number of general approaches and myriad variations of each one. Many aspects of shift systems have been studied, but attempts to reach definitive conclusions about appropriate designs have been hampered by a number of methodological issues. The aim of this systematic review was to provide evidence-based recommendations on the effect of various shift systems on neurobehavioural and physiological functioning and to identify areas which are lacking in appropriate evidence. Two main aspects of shift design were able to be considered-the direction of shift rotation and extended shift length (mainly 12-h shifts). Other areas for which there was at least one relevant paper of adequate methodology were the use of naps during night shifts, the starting time of shifts, and several other specific shift issues. Overall, the review found there is insufficient evidence to support definitive conclusions regarding any of these factors. However, the analysis provides support for the use of forward rotating shift systems in preference to backward rotating shift systems, at last as far as 8-h shifts are concerned. There are many unanswered questions in shift design. For these questions to be answered, it is important that the methodological shortcomings present in most of the studies published to date be overcome. PMID- 17418597 TI - Complete genetic linkage maps from an interspecific cross between Fusarium circinatum and Fusarium subglutinans. AB - The Gibberella fujikuroi complex includes many plant pathogens of agricultural crops and trees, all of which have anamorphs assigned to the genus Fusarium. In this study, an interspecific hybrid cross between Gibberella circinata and Gibberella subglutinans was used to compile a genetic linkage map. A framework map was constructed using a total of 578 AFLP markers together with the mating type (MAT-1 and MAT-2) genes and the histone (H3) gene. Twelve major linkage groups were identified (n=12). Fifty percent of the markers showed significant deviation from the expected 1:1 transmission ratio in a haploid F(1) cross (P <0.05). The transmission of the markers on the linkage map was biased towards alleles of the G. subglutinans parent, with an estimated 60% of the genome of F(1) individuals contributed by this parent. This map will serve as a powerful tool to study the genetic architecture of interspecific differentiation and pathogenicity in the two parental genomes. PMID- 17418598 TI - Assessment of current knowledge of footrot in sheep with particular reference to Dichelobacter nodosus and implications for elimination or control strategies for sheep in Great Britain. AB - A brief summary of the currently accepted aetiology and pathogenesis of footrot and interdigital dermatitis in sheep in Great Britain is presented together with a description of the transmission dynamics of Dichelobacter nodosus. Approaches that may be successful for control or elimination of the disease are proposed and areas of uncertainty in current knowledge highlighted. Research and practice in Great Britain is compared with that from other countries and the proposal made that the environment and/or carrier (non-diseased) sheep may assist persistence of footrot. PMID- 17418599 TI - Is equitation science important to veterinarians? PMID- 17418600 TI - A fourth ventricular ganglioneurocytoma representing with cerebellar epilepsy: a case report and review of the literature. AB - Fourth ventricular low-grade tumoral or dysplastic neuronal lesions have been reported as an epileptic focus for recently described cerebellar epilepsy in the form of repetitive and stereotyped attacks of hemifascial spasm, eye blinking, fascial movements, head deviation and dysautonomic manifestations. The case of a 3-month old infant having fourth ventricular mass with similar symptoms such as paroxysmal facial movements, eye blinking, eyelid contractions and abnormal head posture is reported in this article. After a few days of her admission, her attacks displayed a new form with altered consciousness and left limb jerks which were unresponsive to medical therapy. Following the surgical excision of the lesion 10 months ago, attacks disappeared and she is still seizure free. Histopathological diagnosis was ganglioneurocytoma. The seizures (which may be intractable in cerebellar epilepsy) are thought to have arisen from subcortical structures such as cerebellum, brain stem nuclei or the lesion itself. In the case of intractable episodes, surgical excision may prevent further seizures and help patients have a normal cognitive and motor development. PMID- 17418601 TI - Brain morphology alterations in the basal ganglia and the hypothalamus following prenatal exposure to antiepileptic drugs. AB - If humans are exposed prenatally to antiepileptic drugs (AEDs), cognitive impairment may be the consequence. Driven by results of experimental work showing that AEDs may induce neuronal death in the developing rodent brain, we wanted to explore whether prenatal exposure to AEDs (PAE) may result in structural changes in the human brain. For this purpose we investigated a group of healthy young adults with PAE and a group of age-matched unexposed healthy controls by magnetic resonance imaging (MRI) of the brain. Local differences in cerebral morphology associated with PAE were analysed in volumetric MRI data by use of voxelwise comparisons of grey and white matter images. Significant regional decreases of grey matter volumes were found in PAE subjects in the area of the lentiform nucleus, including both pallidum and putamen bilaterally, and the hypothalamus. No significant regional differences in white matter volumes were found. We conclude that PAE causes subtle morphological changes in grey matter of the human brain which are conform with lower cell numbers in the basal ganglia and the hypothalamus. PMID- 17418602 TI - Three-dimensional QSAR of HPPD inhibitors, PSA inhibitors, and anxiolytic agents: effect of tautomerism on the CoMFA models. AB - The present study was design to examine the effect of tautomerism upon the CoMFA results. Three selected data sets involving protropic tautomerism, which are 21 p hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors, 35 inhibitors of puromycin sensitive aminopeptidase (PSA), and 67 anxiolytic agents, were used for this purpose. Atom-by-atom alignment technique was adopted to superimpose the molecules in the data sets onto a template. The structural alignments using different tautomeric forms had no significant difference except the atoms involved in tautomerism, which ensures, to a great extent, that the differences of the CoMFA results result primarily from the tautomerism. All-orientation and all-placement search (AOS-APS) based CoMFA models, in addition to the conventional ones, were derived for each system and proved to be capable of yielding much improved statistical results. In the cases of the data sets of HPPD inhibitors and PSA inhibitors, excellent AOS-APS CoMFA models (q2>0.8 with four components for the former and q2>0.7 with seven components for the latter) were obtained, and almost no significant difference in statistical quality was observed when using different tautomeric forms to derive the models. However, it was not the case when treating the data set of anxiolytic agents. The keto tautomer, which was the active form of the PBI type inhibitors, produced measurably better results (q2=0.54 with eight components) than that the enol one (q2=0.37 with five components), indicating the importance of selecting proper tautomer in the CoMFA studies. Furthermore, there existed some substantial differences of the electrostatic field contours between the two different tautomeric forms for all of the three systems considered, whereas the differences in the steric field contour maps were limited. This implies that the resulting new potent ligands may be quite different if one utilizes the CoMFA models of different tautomeric forms for guiding further structural refinements. PMID- 17418603 TI - Prediction of factor Xa inhibitors by machine learning methods. AB - Factor Xa (FXa) inhibitors have been explored as anticoagulants for treatment and prevention of thrombotic diseases. Molecular docking, pharmacophore, quantitative structure-activity relationships, and support vector machines (SVM) have been used for computer prediction of FXa inhibitors. These methods achieve promising prediction accuracies of 69-80% for FXa inhibitors and 85-99% for non-inhibitors. Prediction performance, particularly for inhibitors, may be further improved by exploring methods applicable to more diverse range of compounds and by using more appropriate set of molecular descriptors. We tested the capability of several machine learning methods (C4.5 decision tree, k-nearest neighbor, probabilistic neural network, and support vector machine) by using a much more diverse set of 1098 compounds (360 inhibitors and 738 non-inhibitors) than those in other studies. A feature selection method was used for selecting molecular descriptors appropriate for distinguishing FXa inhibitors and non-inhibitors. The prediction accuracies of these methods are 89.1-97.5% for FXa inhibitors and 92.3-98.1% for non-inhibitors. In particular, compared to other studies, support vector machine gives a substantially improved accuracy of 94.6% for FXa non-inhibitors and maintains a comparable accuracy of 98.1% for inhibitors, based-on a more rigorous test with more diverse range of compounds. Our study suggests that machine learning methods such as SVM are useful for facilitating the prediction of FXa inhibitors. PMID- 17418604 TI - Molecular characterization and sex-specific tissue expression of prolactin, somatolactin and insulin-like growth factor-I in yellow perch (Perca flavescens). AB - The cDNA sequences encoding prolactin (PRL), somatolactin (SL) and insulin-like growth factor-I (IGF-I) genes of the yellow perch were obtained. Brain, pituitary, gill, heart, liver, stomach, kidney, spleen, muscle and gonad tissues were analyzed from both male and female adult yellow perch for sex-specific tissue expression. The full length cDNA of yellow perch PRL consists of 2306 bp and PRL expression was highest in the yellow perch pituitary with low to moderate expression in other tissues including brain, gill and post-vitellogenic oocytes. The full length cDNA of yellow perch SL consists of 1589 bp and SL expression was highest in the yellow perch pituitary with low to moderate expression in other tissues including brain, gill, liver, stomach, spleen and kidney. The full length cDNA of yellow perch IGF-Ib consists of 814 bp and tissue expression analysis of yellow perch IGF-I revealed a second yellow perch transcript (IGF-Ia) that is 81 nucleotides smaller. Both IGF-Ib and IGF-Ia had the greatest expression in liver tissue with moderate expression in brain, spleen and kidney tissues of both sexes. These sequences are valuable molecular tools which can be used in future studies investigating the basis for sexually dimorphic growth in yellow perch. PMID- 17418605 TI - Over-expression of IGF-related peptides in stenoses of native arteriovenous fistulas in hemodialysis patients. AB - INTRODUCTION: Insulin-like growth factors (IGFs) are known to play an important role in atherogenesis. The aim of our study was to assess the local expression of IGF-related peptides in stenosed hemodialysis fistulas and compare these with their respective serum levels. METHODS: We investigated 15 stenosed vein segments of primary arteriovenous fistulas, 29 non-stenosed control vein segments from uremic patients and 15 non-stenosed control saphenous vein segments. Immunohistochemistry was performed for IGF-I, insulin, IGF-binding proteins (IGFBPs)-1, -2, -3 and -4, the acid labile subunit (ALS) and type 1 IGF-receptor (IGF-R). Serum levels were measured by specific radioimmunoassays. RESULTS: Compared to both control groups, a significantly higher expression of the following IGF-related peptides was seen in the stenotic (neo)intima: IGF-I, IGFBP 1, -2, -3, -4 and IGF-R; in the stenotic media: IGF-I and IGFBP-3 and in the endothelium of stenotic fistulas: IGF-I (all p<0.05). Staining against ALS and insulin was negative in all vessels. Serum IGF-I levels did not differ. Serum levels of IGFBP-1, -2, -3 and -4 were significantly higher in patients with renal disease (all p<0.05). There were no correlations between local and systemic IGF related peptide levels. There were correlations of neointimal expression of IGF I, IGFBP-1, -2, -3, -4 and IGF-R with both hypercellularity and the presence of inflammatory cells (p<0.05). CONCLUSION: In the stenotic arteriovenous fistula of hemodialysis patients, expression of the peptides IGF-I, IGFBP-1, -2, -3, -4 and IGF-R was significantly increased and showed a positive correlation with neointimal inflammation and hypercellularity (all p<0.05). IGF-related peptides are most likely synthesized locally and might be involved in the initiation and/or progression of neointimal thickening of primary arteriovenous fistulas. PMID- 17418606 TI - Normal adiponectin levels despite abnormal glucose tolerance (or diabetes) and inflammation in adult patients with cystic fibrosis. AB - RATIONALE: Circulating adiponectin levels are negatively associated with glucose intolerance, inflammation and central adiposity. Since these conditions are common in cystic fibrosis (CF), we examined whether adiponectin values are altered in these patients. AIM: To determine if CF patients have altered adiponectin levels and if these levels correlate with glucose tolerance categories (normal, impaired glucose tolerance (IGT) and cystic fibrosis-related diabetes (CFRD)), insulin resistance or inflammatory markers such as fibrinogen and C-reactive protein (CRP). METHODS: Oral glucose tolerance tests (OGTTs) were performed and adiponectin levels were measured in 90 CF patients not known to be diabetic and 15 healthy controls matched for age, sex and body mass index (BMI). Inflammatory markers, serum albumin concentrations and the clinical status of CF patients (i.e. pulmonary function) were also examined. RESULTS: CF pathology was characterized by a high prevalence (43.5%) of glucose tolerance abnormalities: 26.5% of IGT and 17.0% of newly diagnosed CFRD. CF patients also presented systemic inflammation as revealed by a significant increase of fibrinogen (P=0.029) in all patients and higher CRP levels in CFRD patients compared to the controls (P<0.05). On the other hand, CF and control subjects had similar albumin serum concentration. While CF patients and controls had similar serum adiponectin values, women had significantly higher hormone levels than men (P<0.001). Adiponectin levels did not correlate with glucose tolerance, inflammatory markers or insulin resistance. On the other hand, they correlated positively with both total and HDL-cholesterol (P<0.001). CONCLUSION: CF patients did not show any alterations in adiponectin levels despite insulin resistance, glucose intolerance and sub clinical chronic inflammation. Thus, CF appears to be one of the rare conditions in which discordance between adiponectin values and insulin resistance or inflammation is evident. PMID- 17418607 TI - Effects of CB1 antagonist on the control of metabolic functions in obese type 2 diabetic patients. AB - Clinical reports (RIO trials) have shown that chronic administration of a CB cannabinoid receptor antagonist (rimonabant) provides improvements of disturbed metabolic parameters observed in overweight and obese patients with type 2 diabetes. The production of endocannabinoid and the expression of CB1-cannabinoid receptors are largely distributed in the different organs aside from the brain. It is now clearly established that endocannabinoids act both through orexigenic effects and peripheral metabolic effects in various tissues involved in the control of metabolism and energy expenditure (i.e. adipose tissue, liver, gastrointestinal tract, skeletal muscle and pancreas). This review will consider: i) the disturbances of glucose and lipid metabolisms in obese type 2 diabetics; ii) an overview of the pharmacological properties of rimonabant and iii) the various mechanisms involved in tissues and organs to explain the therapeutic efficacy of rimonabant. A special attention will be paid to its utilization in obese type 2 diabetics. The emerging concept of endocannabinoids acting as metabolic regulators is the more likely explanation of the success of rimonabant treatments in phase III studies. PMID- 17418608 TI - [Quality assurance of a virtual simulation software: application to IMAgo and SIMAgo (ISOgray)]. AB - PURPOSE: Virtual simulation process is often used to prepare three dimensional conformal radiation therapy treatments. As the quality of the treatment is widely dependent on this step, it is mandatory to perform extensive controls on this software before clinical use. The tests presented in this work have been carried out on the treatment planning system ISOgray (DOSIsoft), including the delineation module IMAgo and the virtual simulation module SIMAgo. MATERIAL AND METHODS: According to our experience, the most relevant controls of international protocols have been selected. These tests mainly focused on measuring and delineation tools, virtual simulation functionalities, and have been performed with three phantoms: the Quasar Multi-Purpose Body Phantom, the Quasar MLC Beam Geometry Phantom (Modus Medical Devices Inc.) and a phantom developed at Hospital Tenon. RESULTS: No major issues have been identified while performing the tests. These controls have emphasized the necessity for the user to consider with a critical eye the results displayed by a virtual simulation software. The contrast of visualisation, the slice thickness, the calculation and display mode of 3D structures used by the software are many factors of uncertainties. CONCLUSION: A virtual simulation software quality assurance procedure has been written and applied on a set of CT images. Similar tests have to be performed periodically and at minimum at each change of major version. PMID- 17418609 TI - Monarch-1/PYPAF7 and other CATERPILLER (CLR, NOD, NLR) proteins with negative regulatory functions. AB - CATERPILLER is a mammalian gene family with signature NBD and LRR domains. Several members of this family are positive regulators of inflammatory responses. Others, however, exert negative effects on proinflammatory responses. These data are particularly convincing when shRNA/siRNA are used. This review focuses on the Monarch-1/PYPAF7 gene with brief discussions of CLR16.2/NOD3, PYPAF2/PAN1/NALP2, and PYPAF3. PMID- 17418610 TI - Malaria transmission-blocking vaccines: the bonus of effective malaria control. AB - Reduction of transmission is important for effective malaria control. Standard microscopic detection grossly underestimates the human transmission reservoir. Malaria transmission intensity is defined by local and focal conditions. Transmission-blocking vaccines may be effective by directly targeting identified hot spots of transmission. PMID- 17418611 TI - Cancer modeling: modern imaging applications in the generation of novel animal model systems to study cancer progression and therapy. AB - Cancer is the result of a series of genetic and epigenetic mutations that evolve over years even decades and lead to the transformed phenotype. Paradoxically, most methods developed to study these changes are static and do not provide insights on the dynamics of the sequela of steps involved in tumorigenesis. This major shortcoming now can be overcome with the application of reporter genes and imaging technologies, which are providing tools to examine specific molecular events and their role in the carcinogenic process in single cells. In the last decade reporter-based biosensors were created to study gene transcription, protein/protein interactions, sub-cellular trafficking and protease activities; this wealth of systems enable to monitor intracellular signaling pathways at several key check points specifically involved in cancer cell development. The challenge is now to extend cell-based models to the generation of reporter mice, where non-invasive in vivo imaging technologies allow to follow single molecular events. When combined with murine models of cancer, these technologies will give an unprecedented opportunity to spatio-temporally investigate the molecular events resulting in neoplasia. The aim of the present review is to highlight the major changes occurring in this rapidly evolving field and their potential for increasing our knowledge in cancer biology and for the research of novel and more efficacious therapies. PMID- 17418612 TI - Clinical analysis of salivary gland tumor cases in West China in past 50 years. AB - In our study, 3461 cases of salivary gland tumor treated between 1955 and 2002 at West China Stomatology Hospital of Sichuan University were retrospectively analyzed, and compared with the previous reports. Measures such as age, tumor location, tumor histological type, and the nature of the growth (benign or malignant) were recorded at the same time. The findings are as follows: the average ages of salivary gland tumor patients were 41.38 years for the benign cases and 45.20 for the malignant ones; the male:female ratio was 0. 99:1 in the benign cases and 1.34:1 in the malignant ones; primary tumors were mostly in the parotid gland, palate and submandibular gland in sequence. Pleomorphic adenoma was the most frequent benign tumor followed by Warthin's tumor and basal cell adenoma, whereas mucoepidermoid carcinoma, adenoid cystic carcinoma and adenocarcinoma not otherwise specified were the most frequent malignant tumors. The incidence of salivary gland tumors increased with age. The male:female ratio of malignant tumors was higher than that of benign ones. The parotid gland and palate were the most common locations of salivary gland tumors. Pleomorphic adenoma and mucoepidermoid carcinoma were the most frequent benign and malignant tumors, respectively. PMID- 17418613 TI - Glutathione S-transferase polymorphisms and oral cancer: a case-control study in Rio de Janeiro, Brazil. AB - This study evaluates the influence of genetic polymorphisms at GSTM1, GSTT1 and GSTP1 gene loci on oral cancer susceptibility among Brazilians from Rio de Janeiro. DNA extracted from white blood cells of 231 oral cancer patients and 212 hospital controls was analyzed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) methods. GSTM1 polymorphism distribution was different between cases and controls (P=0.006), with an overrepresentation of GSTM1 A/B genotype in controls. GSTM1 A/B individuals were at decreased oral cancer risk (OR=0.08; 95% CI=0.05-0.62). No statistically significant association was observed for GSTT1 and GSTP1 polymorphisms. Differences in the GSTM1 and GSTT1 null genotype frequencies were observed between individuals of European origin and African origin, but these genotypes do not seem to influence the risk of oral cancer. Therefore, these results do not support the hypothesis of increased risk of GSTP1 G/G, GSTM1 or GSTT1 null genotypes for developing cancer in oral cavity, but the GSTM1 A/B genotype emerged as a protective factor. PMID- 17418614 TI - Lower prevalence but favorable survival for human papillomavirus-related squamous cell carcinoma of tonsil in Taiwan. AB - Human papillomavirus (HPV) is frequently detected in squamous cell carcinoma of tonsil (TSCC) among the Western population. However, the only reported study on Chinese patients in the English literature demonstrated absence of HPV in TSCC. To evaluate the prevalence and clinical significance of HPV-related TSCC in Taiwan, we performed PCR with MY09/MY11 consensus primers and in situ hybridization to study 111 TSCC samples. The results showed that only 12.6% TSCC were HPV-positive. The favorable 5-year survival rate correlated significantly with HPV positivity (p=0.007), female (p=0.046), and early tumor (T) stage (p<0.001), but Cox's regression analysis revealed that only the status of HPV (p=0.04) and T stage (p=0.004) were independent prognostic factors for survival. In conclusion, the prevalence of HPV-related TSCC is much lower in Taiwan comparing with the Western population, and the prognosis of HPV-positive TSCC is better than that of HPV-negative TSCC. PMID- 17418615 TI - Lower-extremity lymphedema following neck dissection--an uncommon complication after cervical ligation of the thoracic duct. AB - Thoracic duct injuries and chylous fistula are well-known complications of neck dissection, occurring in 1-2% of cases. Management of these injuries can be conservative or operative. Conservative treatment consists of fat restricted diet or total parenteral nutrition reducing the volume of chyle production. Operative management includes exploration of the neck or if necessary open thoracotomy to ligate the thoracic duct. Following cervical thoracic duct ligation only few complications like chylothorax or chylous ascites are described in the literature. To the best authors knowledge, this is the first report in the english literature describing lower-extremity lymphedema following cervical thoracic duct ligation. PMID- 17418616 TI - Estradiol, tamoxifen and ICI 182,780 alter alpha3 and beta1 integrin expression and laminin-1 adhesion in oral squamous cell carcinoma cell cultures. AB - Adhesion molecules such as integrins and extracellular matrix proteins like laminins have been identified to play an important role in cell proliferation, migration and invasion by regulating cell-extracellular matrix interaction in various cancers including oral squamous cell carcinoma (OSCC). In this study, the effect of estradiol (E2), and the E2 antagonists tamoxifen (TAM) and ICI 182,780 (ICI) on the expression of integrins and adhesion to laminin-1 in different OSCC in vitro models was analyzed. TAM and ICI inhibited growth in all OSCC cell lines. Dependent on estrogen receptor (ER) status E2 displayed a significant influence on growth after long-term administration. ICI reduced laminin-1 adhesion in all cell lines. beta1 Integrin transcription is reduced with TAM and E2 and alpha3 cell surface expression with TAM. This study shows that OSCC is estrogen and SERM sensitive and that these compounds can modulate cell-matrix interaction in part by modulating integrin expression and translation. The investigation also confirms that growth is significantly influenced by these adjuvant therapeutics. These data suggest that a greater understanding of basic biology and mechanisms of the ER and its ligands in oral squamous cells is needed to elucidate the use of specific pharmacological agents as therapeutics of anti tumorigenic pathways. PMID- 17418617 TI - Short tandem repeat polymorphism in exon 4 of esophageal cancer related gene 2 predicts relapse of oral squamous cell carcinoma. AB - Short tandem repeat (STR) polymorphisms in exon 4 of esophageal cancer related gene 2 (ECRG2) are a prognostic marker for squamous cell carcinoma (SCC) of the esophagus. The aim of the present study was to correlate these STRs with clinical outcome of the similar tumor type oral squamous cell carcinoma (OSCC). DNA of 81 patients that underwent complete surgical resection of OSCC was analyzed for STRs TCA3/TCA3, TCA3/TCA4 and TCA4/TCA4 in exon 4 of ECRG2 by PCR, capillary electrophoresis and DNA sequencing. ECRG2 STR TCA3/TCA3 were found in 45 (56%), TCA3/TCA4 in 33 (41%) and TCA4/TCA4 in 3 (3%) patients. TCA3/TCA3 was significantly associated with reduced relapse-free survival of OSCC, compared with TCA3/TCA4 and TCA4/TCA4 genotypes (P<0.05; log-rank test). TCA3/TCA3 STR was independent prognostic factor determined by multivariate Cox regression analysis (p<0.05). STR polymorphism TCA3/TCA3 in exon 4 of ECRG2 is associated with poor relapse-free survival in surgically completely resected OSCC patients and might be a potential prognostic marker. PMID- 17418618 TI - EpCAM expression in squamous cell carcinoma of the oral cavity: frequency and relationship to clinicopathologic features. AB - This retrospective study was designed to investigate the prognostic significance of EpCAM expression in human oral squamous cell carcinoma on a long-term follow up. EpCAM expression was examined immunohistochemically on a tissue microarray (TMA) of paraffin embedded tissue specimens from 77 consecutive patients who underwent surgical treatment for squamous cell carcinoma of the oral cavity in the period between 1980 and 1997 at the Department of Craniomaxillofacial and Oral Surgery, Innsbruck Medical University. High EpCAM expression was found in 17 (22.1%) of the tumor samples. Using Kaplan-Meier analysis no correlation of EpCAM overexpression was observed with conventional clinicopathologic features in this patient cohort. Taken together, these data suggest that EpCAM might become an attractive treatment target in a subgroup of patients with OSCC. PMID- 17418620 TI - The upregulation of metallothionein-1 expression in areca quid chewing-associated oral squamous cell carcinomas. AB - Metallothioneins (MTs) are a family of low molecular weight, cysteine-rich, inducible, intracellular proteins that bind heavy metals with high affinity. MT-1 is known as a stress-inducible protein and functions as an antioxidant enzyme. Areca quid chewing is a major risk factor in the development and further progression of oral squamous cell carcinoma (OSCC). The aim of this study was to compare MT-1 expression in normal human oral epithelium and OSCC and further explore the potential mechanism that may lead to induce MT-1 expression. Thirty four OSCC and 10 normal epithelium specimens were examined by immunohistochemistry and analyzed by the clinico-pathological profiles. The oral epithelial cell line GMN cells were challenged with arecoline, a major areca nut alkaloid, by reverse-transcriptase polymerase chain reaction. Furthermore, tobacco smoke carcinogen benzo[a]pyrene (BaP) and glutathione (GSH) precursor N acetyl-l-cysteine (NAC) were added to find the possible regulatory mechanisms. The results from immunohistochemistry demonstrated that MT-1 expression was significantly higher in OSCC specimens (p<0.05). No significant difference in MT 1 expression was observed with respect to age, sex, T category, and stage (p>0.05). The high MT-1 expression was associated with lymph node metastasis (p=0.012). In addition, arecoline was found to elevate MT-1 mRNA in a dose dependent manner (p<0.05). Furthermore, the addition of BaP enhanced the arecoline-induced MT-1 expression (p<0.05). The addition of NAC markedly inhibited the arecoline-induced MT-1 expression (p<0.05). These results lead to the conclusion that MT-1 expression is significantly upregulated in areca quid chewing associated-OSCC. The expression profile suggests MT-1 could be used clinically as a marker for tumors possessing the potential for lymph node metastasis. The compounds of tobacco products may act synergistically in the pathogenesis of OSCC in areca quid chewers. The regulation of MT-1 expression induced by arecoline is critically dependent on the intracellular GSH concentration. PMID- 17418619 TI - Expression of novel p53 isoforms in oral lichen planus. AB - Oral lichen planus (OLP) is a chronic inflammatory disease of unknown origin, showing little spontaneous regression. WHO classifies OLP as a premalignant condition, however, the underlying mechanisms initiating development of cancer in OLP lesions are not understood. The p53 tumour suppressor plays an important role in many tumours, and an increased expression of p53 protein has been seen in OLP lesions. Recently it was shown that the human TP53 gene encodes at least nine different isoforms. Another member of the p53 family, p63, comprises six different isoforms and plays a crucial role in the formation of oral mucosa, salivary glands, teeth and skin. It has also been suggested that p63 is involved in development of squamous cell carcinoma of the head and neck (SCCHN). In contrast to p53, a decreased expression of p63 protein has been seen in OLP lesions. In this study, we mapped the expression of five novel p53 isoforms at RNA and protein levels in OLP and matched normal controls. In the same samples we also measured levels of p63 isoforms using quantitative RT-PCR. Results showed p53 to be expressed in all OLP lesions and normal tissues. The p53 beta and delta 133p53 isoforms were expressed in the majority of samples whereas the remaining three novel isoforms analysed were expressed in only a few samples. Levels of p63 isoforms were lower in OLP lesions compared with normal tissue, however, changes were not statistically significant. PMID- 17418622 TI - Crystal growth and characterization of K[CS(NH2)2]4Br--A semiorganic non-linear optical crystal. AB - This paper presents the synthesis and crystal growth of optically non-linear semi organic crystal potassiumtetrakisthioureabromide (KTTB) using water and a mixture of acetone and water (1:1) by slow evaporation of aqueous solution at room temperature. The solubility has been determined in both the solvents at various temperatures. The compound crystallizes in non-centrosymmetric space group P41. The crystalline perfection of the grown crystals has been analysed by high resolution X-ray diffraction measurements. The IR and polarized Raman spectra have been interpreted using both the factor group analysis. The results of the correlation field splitting of the thiourea internal vibrations are discussed. The optical characterization of the crystal shows the cut off wavelength near the UV region. Also the crystal shows good propensity to emit second harmonic generation of Nd:YAG laser. PMID- 17418623 TI - Theoretical study of the molecular structure and normal coordinate analysis of hydrogen cyanide addition compound with boron trifluoride, HCN-BF(3). AB - An extensive HF, MP2, B3LYP and CCSD study of the molecular structure and normal vibrations have been performed for the HCN-BF(3) molecule. Calculations with a wide range of basis sets were classified into two groups based on the optimized N B bond distance. The results for Group A are compared with the experimental structure of the solid phase molecules. The N-B lengths of Group A are approximately linear related to the N-B-F valence angles and also to the N-B stretching frequencies. HF/DZV calculation was used to represent the solid phase model. The N-B lengths of Group B are close to those of the gas phase molecule and both N-B-F angles and N-B sensitive frequencies have roughly the same values. Differences in the chemical bond between gaseous and solid phase HCN-BF(3) are discussed based on the calculated force constants, vibrational frequencies and potential energy distributions. Vibration mode analysis indicates that the nu(4) mode in the 600-700 cm(-1) region can be assigned to the BF(3) symmetric deformation, which shifts upon (10)B/(11)B isotopic substitution. The nu(5) mode which is insensitive to isotope substitution and changes band position with the N B distance is assigned to the N-B bond stretching vibration. PMID- 17418624 TI - Theoretical calculation of EPR g factors for Ni(3+) ion at the interstitial site of SnO(2) crystal. AB - The EPR g factors g(x), g(y) and g(z) for Ni(3+) ion at the interstitial site in the rutile-type SnO(2) crystal are calculated from the second-order perturbation formulas of g factors based on the cluster approach for 3d(7) ion in rhombic symmetry. The calculated results are in agreement with the experimental values. The local lattice distortion induced by the impurity Ni(3+) at the interstitial site of SnO(2) is also estimated from the calculations. These results are discussed. PMID- 17418625 TI - Spectroscopic evaluation of Co(II), Ni(II) and Cu(II) complexes derived from thiosemicarbazone and semicarbazone. AB - Co(II), Ni(II) and Cu(II) complexes were synthesized with thiosemicarbazone (L(1)) and semicarbazone (L(2)) derived from 2-acetyl furan. These complexes were characterized by elemental analysis, molar conductance, magnetic moment, mass, IR, electronic and EPR spectral studies. The molar conductance measurement of the complexes in DMSO corresponds to non-electrolytic nature. All the complexes are of high-spin type. On the basis of different spectral studies six coordinated geometry may be assigned for all the complexes except Co(L)(2)(SO(4)) and Cu(L)(2)(SO(4)) [where L=L(1) and L(2)] which are of five coordinated square pyramidal geometry. PMID- 17418626 TI - Simpler spectrophotometric assay of paracetamol in tablets and urine samples. AB - A very fast, economical and simpler direct spectrophotometric method was investigated for paracetamol (PC) determination in aqueous medium without using any chemical reagents. The method is based on the photo-absorption of the analyte at 243 nm after dissolution in water. The change in structure of PC after addition of water was studied by comparing the corresponding FTIR spectra. Optimization studies were conducted by using a 5 microg ml(-1) standard solution of the analyte. Various parameters studied include, time for stability and measurement of spectra, effect of HCl, NaOH, CH(3)COOH and NH(3) for change in absorbance and shift in spectra, interference by some analgesic drugs and some polar solvents and temperature effect. After optimization, Beer's law was obeyed in the range of 0.3-20 microg ml(-1) PC solution with a correlation coefficient of 0.9999 and detection limit of 0.1 microg ml(-1). The newly developed method was successfully applied for PC determination in some locally available tablets and urine samples. The proposed method is very useful for quick analysis of various types of solid and liquid samples containing PC. PMID- 17418627 TI - Vibrational spectroscopy investigation using ab initio and density functional theory analysis on the structure of 3,4-dimethylbenzaldehyde. AB - The FT-IR and FT-Raman spectra of 3,4-dimethylbenzaldehyde (3,4-DMB) has been recorded in the region 4000-400 and 3500-100 cm(-1), respectively. The optimized geometry, frequency and intensity of the vibrational bands of 3,4-DMB were obtained by the ab initio and DFT levels of theory with complete relaxation in the potential energy surface using 6-311G(d,p) basis set. The harmonic vibrational frequencies were calculated and the scaled values have been compared with experimental FT-IR and FT-Raman spectra. The observed and the calculated frequencies are found to be in good agreement. The experimental spectra also coincide satisfactorily with those of theoretically constructed bar type spectrograms. PMID- 17418628 TI - Syntheses, characterization and fluorescent properties of two triethylene-glycol dicoumarin-3-carboxylates. AB - Two novel triethylene-glycol dicoumarin-3-carboxylates were synthesized and characterized by element analysis, (1)H NMR, FT-IR and UV-vis absorption spectra. The photoluminescent behaviors of triethylene-glycol dibenzo[5,6]coumarin-3 carboxylate doped in PMMA were discussed. These compounds exhibit strong blue emission under ultraviolet light excitation. Compared to coumarin-3-triethylene glycol diester, the absorption and emission spectra of benzo[5,6]coumarin-3 triethylene glycol diester was bathochromically shifted due to the larger conjugation of benzene moiety. These derivatives have potential possible to explore organic electroluminescent materials. PMID- 17418629 TI - Selective fluorescent Hg(II) detection in aqueous solutions with a dye intermediate. AB - A dye intermediate, 1-amino-8-naphthol-3,6-disulfonic acid sodium (ANDS) was first used to selectively recognize Hg(II) in aqueous solutions with its fluorescence being strong quenched. The fluorescence quenching of ANDS was attributed to the formation of an inclusion complex between Hg(II) and ANDS by 2:1 complex ratio (K=6.2 x 10(9)), which has been utilized as the basis of the fabrication of the Hg(II)-sensitive fluorescent chemosensor. The analytical performance characteristics of the proposed chemosensor were investigated. The sensor shows a linear response toward Hg(II) in the concentration range 2.9 x 10( 6) to 5.5 x 10(-5)M with a limit of detection of 5.3 x 10(-7)M, and a working pH range from 5.0 to 9.0. It shows excellent selectivity for Hg(II) over a large number of cations such as alkali, alkaline earth and transitional metal ions. The proposed method was utilized successfully for the detection of Hg(2+) in water samples. PMID- 17418630 TI - FT-Raman and infrared spectra and vibrational assignments for 3-chloro-4 methoxybenzaldehyde, as supported by ab initio, hybrid density functional theory and normal coordinate calculations. AB - Fourier-transform laser Raman (3500-50 cm(-1)) and infrared (4000-400 cm(-1)) spectral measurements have been made for the solid 3-chloro-4 methoxybenzaldehyde. The electronic structure calculations -ab initio (RHF) and hybrid density functional methods (B3LYP and B3PW91) -- have been performed with 6-31G* and 6-311G* basis sets. Molecular electronic energies, equilibrium geometries, IR and Raman spectra have been computed. Potential energy distribution (PEDs) and normal mode analysis have also been performed. A complete assignment of the observed spectra has been proposed. Investigation of the relative orientation of the aldehydic oxygen and chlorine atom with respect to the methoxy group has shown that two forms, O-cis and O-trans exist, with O-trans form being more stable. The energy difference between O-cis and O-trans forms is 0.057 kcal/mol (21 cm(-1)) with B3LYP/6-31G*, which is less than the calculated torsional vibrational frequencies of the aldehyde and methoxy group. In the CH (O) aldehydic stretching region five observed bands are probably due to multiplet Fermi resonance. An infrared doublet near 1700 cm(-1) with nearly equal intensities has been ascribed to the Fermi resonance: the two bands at 1696 and 1679 cm(-1) arise due to the interaction between the CO stretching fundamental and a combination of O-CH(3) and CC stretching vibrations. PMID- 17418631 TI - The molecular structure and vibrational spectra of 2-chloro-N (diethylcarbamothioyl)benzamide by Hartree-Fock and density functional methods. AB - 2-Chloro-N-(diethylcarbamothioyl)benzamide (C(12)H(15)ClN(2)OS) has been synthesized and characterized by elemental analysis and IR spectroscopy. The crystal and molecular structure of the title compound has been determined from single crystal X-ray diffraction data. It crystallizes in the orthorhombic space group Pbca, Z=8 with a=9.581(3)A, b=9.992(3)A, c=26.640(8)A, V=2550.5(13)A(3) and D(calc)=1.410 Mg/m(3). The molecular geometry and vibrational frequencies of 2 chloro-N-(diethylcarbamothioyl)benzamide in the ground state have been calculated using the Hartree-Fock and density functional using Becke's three-parameter hybrid method with the Lee, Yang and Parr correlation functional (B3LYP) methods with 3-21G and 6-31G(d) basis sets. The calculated geometric parameters were compared to the corresponding X-ray structure of the title compound. The raw B3LYP frequencies approximate the experimental data much better than the results of Hartree-Fock. The scaled B3LYP/6-31G(d) results were more reliable than those obtained using the B3LYP/3-21G method with the mean absolute deviation about 13.7 cm(-1). On the basis of the comparison between calculated and experimental results, assignments of fundamental vibrational modes were examined. PMID- 17418632 TI - Investigations of the EPR zero-field splitting and the defect structure for Mn(2+) and Fe(3+) ions in anatase crystals. AB - The EPR zero-field splittings D of Mn(2+) and Fe(3+) ions in anatase crystals at room and low temperatures are calculated from the high-order perturbation formula of zero-field splitting D for 3d(5) ions in tetragonal symmetry based on the dominant spin-orbit coupling mechanism. The calculated results are consistent with the observed values. From the calculations, Mn(2+) and Fe(3+) ions are suggested to substitute for Ti(4+) ions in anatase (in the previous paper, Mn(2+) ion was suggested at an interstitial site rather than substitutional site) and the defect structures (characterized mainly by the local oxygen parameter u) for both tetragonal Mn(2+) and Fe(3+) impurity centers are estimated. The different zero-field splitting at room and low temperatures are due mainly to the change of local oxygen parameter u with the temperature. These results are discussed. PMID- 17418633 TI - Synthesis, characterization of Ag(I), Pd(II) and Pt(II) complexes of a triazine-3 thione and their interactions with bovine serum albumin. AB - Ag(I), Pd(II) and Pt(II) complexes of 5-methoxy-5,6-diphenyl-4,5-dihydro-2H [1,2,4]triazine-3-thione (LH(2)OCH(3)) have been synthesized and characterized by elemental analysis, molar conductance, (1)H NMR, IR spectra, UV spectra and thermal analysis (TG-DTA). The components of the three complexes are [Ag(C(15)H(10)N(3)S)](6), Pd(C(15)H(10)N(3)S)(2) and Pt(C(15)H(10)N(3)S)(2).C(3)H(6)O.2H(2)O, respectively. All the complexes are nonelectrolyte and have high thermodynamic stability. The ligand may act as bidentate NS donor for Pd(II) and Pt(II) complexes, while it seems to be bidentate NS bridging via sulphur atom for Ag(I) complex. A planar quadrangular structure is proposed for Pd(II) and Pt(II) complexes and Ag(I) complex may be a hexanuclear cluster. Their interactions with bovine serum albumin (BSA) are investigated using steady state fluorescence technology. It is observed that all of them can quench the intrinsic fluorescence of BSA through static quenching procedure. The binding constants (K(A)) at different temperatures, thermodynamic parameters enthalpy changes (DeltaH) and entropy changes (DeltaS) between BSA and the compounds are calculated. Based on the values of DeltaH and DeltaS, it is judged that the main acting force of PtL(2).C(3)H(6)O.2H(2)O with BSA may be electrostatic interaction, and for the LH(2)OCH(3), Ag(6)L(6) and PdL(2), hydrophobic and electrostatic interactions may be involved in their binding processes. PMID- 17418634 TI - Comparative study on digestive lipase activities on the self emulsifying excipient Labrasol, medium chain glycerides and PEG esters. AB - Labrasol is a lipid-based self-emulsifying excipient used in the preparation of lipophilic drugs intended for oral delivery. It is mainly composed of PEG esters and glycerides with medium acyl chains, which are potential substrates for digestive lipases. The hydrolysis of Labrasol by porcine pancreatic extracts, human pancreatic juice and several purified digestive lipases was investigated in the present study. Classical human pancreatic lipase (HPL) and porcine pancreatic lipase, which are the main lipases involved in the digestion of dietary triglycerides, showed very low levels of activity on the entire Labrasol excipient as well as on separated fractions of glycerides and PEG esters. On the other hand, gastric lipase, pancreatic lipase-related protein 2 (PLRP2) and carboxyl ester hydrolase (CEH) showed high specific activities on Labrasol. These lipases were found to hydrolyze the main components of Labrasol (PEG esters and monoglycerides) used as individual substrates, whereas these esters were found to be poor substrates for HPL. The lipolytic activity of pancreatic extracts and human pancreatic juice on Labrasol(R) is therefore mainly due to the combined action of CEH and PLRP2. These two pancreatic enzymes, together with gastric lipase, are probably the main enzymes involved in the in vivo lipolysis of Labrasol taken orally. PMID- 17418635 TI - PPARgamma in immunity and inflammation: cell types and diseases. AB - The lipid activated transcription factor, PPARgamma appears to have multiple functions in the immune system. There are several cell types expressing the receptor, most prominently antigen presenting cells, such as macrophages and dendritic cells. The receptor's activation leads to primary transcriptional activation of many, mostly lipid metabolism-related genes. However, gene regulation also occurs on immunity and inflammation-related genes. Key questions are: in what way lipid metabolism and immune regulation are connected and how activation and/or repression of gene expression may modulate inflammatory and anti-inflammatory responses and in what way can these be utilized in therapy. Here we provide a cell type and disease centric review on the role of this lipid activated transcription factor in the various cells of the immune system it is expressed in, and in some major inflammatory diseases such as atherosclerosis, inflammatory bowel disease and rheumatoid arthritis. PMID- 17418636 TI - Peak alignment using restricted edit distances. AB - A peak is a pair of real values (x,y), where x is the time when peak of height y is registered. In the peak alignment problem, we are given two sequences of peaks, and our task is to align the sequences allowing some basic edit operations on the peaks. We study an instance of the peak alignment problem that arises in the analysis of Mass Spectrometry data in Systems Biology. There the measurement technique guarantees that two peaks (x,y), (x',y') can only be considered the same if x is close enough to x', and y is close enough to y'. We review some methods to do alignment under such restrictions on matches. PMID- 17418637 TI - Role of insertion elements and yycFG in the development of decreased susceptibility to vancomycin in Staphylococcus aureus. AB - The glycopeptide antibiotic vancomycin acts by binding to the D-alanyl-D-alanine terminus of the cell wall precursor lipid II in the cytoplasmic membrane. The purpose of this study was the identification of genes that might be involved in the vancomycin resistance mechanism. To this end, the expression profiles of two vancomycin intermediately resistant Staphylococcus aureus (VISA) strains, the clinical isolate S. aureus SA137/93A (Etest: 8 microg/ml) and its laboratory mutant S. aureus SA137/93G (Etest: 12 microg/ml) were analyzed using an S. aureus full-genome chip. The results indicated that an essential two-component regulatory system, yycF (vicR) and yycG (vicK) was drastically up-regulated in strain SA137/93A. Sequencing of the yycFG promoter region of strain SA137/93A revealed an insertion of IS256 in the predicted promoter region creating a potentially stronger hybrid promoter. In strain SA137/93G, IS256 was not integrated in the yycFG promoter region but, in previous studies, a copy of IS256 had been found to inactivate the tcaA gene (Maki et al. Antimicrob. Agents and Chemother. 48, 1953-1959 (2004)). Detailed population analyses showed that, in addition to the loss of SCCmec, the inactivation of tcaA seems to cause at least part of the increase in teicoplanin and vancomycin resistance in strain SA137/93G. PMID- 17418638 TI - Physiological consequences of a high work of breathing during heavy exercise in humans. AB - The healthy respiratory system has a remarkable capacity for meeting the metabolic demands placed upon it during strenuous exercise. For example, in order to regulate alveolar partial pressure of oxygen and carbon dioxide during heavy workloads, a 20-fold increase in alveolar ventilation can occur. The high metabolic costs and subsequent increased work of breathing associated with this ventilatory increase can result in a number of limitations to the healthy respiratory system. Two examples of respiratory system limitations that are associated with a high work of breathing are expiratory flow limitation and exercise-induced diaphragmatic fatigue. Expiratory flow limitation can lead to an inability to increase alveolar ventilation (V (A)) in the face of increasing metabolic demands, resulting in gas exchange impairment and diminished endurance exercise performance. Furthermore, the high ventilatory requirements of endurance athletes and the inherent anatomical differences in females could make these groups more susceptible to expiratory flow limitation. Fatigue of the diaphragm has also been documented after strenuous exercise and may be related to a mechanism which increases sympathetic vasoconstrictor outflow and reduces limb blood flow during prolonged exercise. This competition between the muscles of respiration and locomotion for a limited cardiac output may have dramatic consequences for exercise performance. This brief review summarizes the literature as it pertains to the work of breathing, expiratory flow limitation, and exercise-induced diaphragmatic fatigue in healthy humans. PMID- 17418639 TI - Immunomodulation in the pathogenesis of Bordetella pertussis infection and disease. AB - Bordetella pertussis infection of the airways causes the disease pertussis (or whooping cough). The infection can be fatal in infants, but in older children, adolescents and adults usually results in a chronic cough of varying severity that persists long after clearance of the infection. The cause of the cough is unknown, but is presumably a result of the pathogenic effects of one or more of the various virulence factors produced by this bacterium. Accumulating recent evidence indicates that the majority of the virulence-associated effects of these factors is devoted to suppression and modulation of the host immune response, which can be skewed towards the recently described Th17 profile. Although the interplay between virulence factors and immune mechanisms might have evolved to benefit both partners in the host-pathogen interaction, it could also contribute to the severe disease pathology associated with this infection. PMID- 17418640 TI - Opportunistic infections in lung disease: Pseudomonas infections in cystic fibrosis. AB - Pseudomonas aeruginosa is an opportunistic pathogen that significantly contributes to morbidity and mortality in patients with cystic fibrosis. Defective mucociliary clearance associated with the absence of the functional cystic fibrosis transmembrane conductance regulator in airway epithelium plays a critical role in the initial colonization of this pathogen. P. aeruginosa, while initiating a profound inflammatory response, employs multiple mechanisms to evade immune clearance. The capacity to grow in biofilms and the selection of mutants with a mucoid phenotype are major adaptations that allow its persistence in the airways. PMID- 17418641 TI - The prognosis of latent tuberculosis: can disease be predicted? AB - In humans, Mycobacterium tuberculosis persists for long periods in a clinically latent state, creating a huge reservoir of 'silent' tuberculosis (TB) (roughly one-third of the global population) from which new cases continually arise. A prognostic marker for active TB would enable targeted treatment of the small fraction of infected individuals who are most at risk of developing contagious TB, contributing greatly to TB control efforts. Here, we propose that TB-specific interferon-gamma release assays might be useful for identifying individuals with progressive infections who are likely to develop the disease. This might provide an unprecedented advantage for TB control, namely targeted preventive therapy for individuals who are most at risk of developing active contagious TB. PMID- 17418642 TI - Do clinical guidelines enhance safe practice in obstetrics and gynaecology? AB - Clinical guidelines are increasingly used to promote a more uniform standard of high-quality evidence-based health care. International agencies advocate guideline development methods founded on three principles: that recommendations are evidence-based, are explicitly linked to the type and quality of evidence, and are developed by multidisciplinary stakeholder groups. Numerous interventions have been described to support the implementation of guidelines. Systematic reviews suggest that most interventions produce modest to moderate improvements in care; multifaceted interventions appear to be no more effective than single interventions, and the lowest-cost implementation strategy (dissemination of printed materials) may improve care and be feasible in many settings. Given the considerable costs of developing valid guidelines de novo, we advocate local adaptation of existing guidelines if available. We suggest a pragmatic framework to assist policy-makers and clinicians in deciding how best to use the scarce resources available for quality-improvement activities. PMID- 17418643 TI - Subacute postictal aggression in patients with epilepsy. AB - Three men with epilepsy (age range, 38-62) who exhibited brief episodes of violent behavior during the postictal period are described. Disease duration ranged from 27 to 44 years. Patients had both complex partial seizures and secondarily generalized tonic-clonic seizures, which were refractory to antiepileptic drugs. Postictal aggression occurred shortly after a seizure and lasted 5-30 minutes. The patients displayed physically and verbally aggressive behavior toward others, but regained consciousness promptly and showed regret afterward. Interictal EEGs revealed temporal spikes, SPECT showed hypoperfusion in the temporal and frontal areas in two patients, and neuropsychological examination revealed poor frontal lobe function in two patients. Characteristics of our cases are consistent with subacute postictal aggression (SPA) reported previously. Epilepsy of prolonged duration and brain dysfunction involving a broad area including the temporal and frontal lobes may be associated with the occurrence of subacute postictal aggression. PMID- 17418644 TI - How reliable is early treatment response in predicting long-term seizure outcome? AB - Assessing if an early therapeutic response to an antiepileptic drug (AED) is of prognostic value for long-term outcome is of considerable clinical interest. To that end, we evaluated the likelihood that a patient who becomes seizure-free after 6 months of treatment with a single AED would lose that response at 12 months, or vice versa. In a post hoc analysis, data from five comparative, double blind, single-drug studies designed to evaluate the efficacy of treatment of patients with partial seizures with oxcarbazepine versus carbamazepine, phenobarbital, phenytoin, and valproate for approximately 1 year were pooled to investigate same-patient seizure outcome at 6 and 12 months. These studies had similar dosing regimens and included a wide range of patients with newly diagnosed and chronic epilepsy. The main finding in the population included in the analysis is that those patients who are seizure-free at 6 months have a 90% chance of being seizure-free at 12 months, whereas those not seizure-free at 6 months have only a 45% chance of being seizure-free at 12 months (chi(2)=118.716, P<0.000001, odds ratio=11.23 with 95% confidence limits 6.8-18.7). In a worst case assessment, those not seizure-free at 6 months have only an 18% chance of being seizure-free at 12 months (chi(2)=408.105, P<0.000001, odds ratio=41.23 with 95% confidence limits 26.4-65.85). Failure to maintain the response in 10% of patients, including 4% with two or more seizures, was noted with all AEDs studied here and in patients with newly diagnosed as well as chronic epilepsy. Among patients with seizures in Months 1-6, those with newly diagnosed epilepsy became seizure-free more often over time than those with chronic epilepsy. The main conclusion is that response at 6 months is an excellent predictor of response at 12 months. PMID- 17418645 TI - Nonconvulsive status epilepticus manifesting as pure alexia (alexia without agraphia). AB - Pure alexia is a rare disorder usually caused by an ischemic or hemorrhagic stroke. We describe a case of pure alexia due to nonconvulsive status epilepticus (NCSE). This 57-year-old man presented in a hyperosmolar, nonketotic state with fluctuations in mental status and an inability to read. His EEG was diagnostic of NCSE originating from the left temporo-occipital region. MRI of the brain revealed increased FLAIR signal over the left occipitotemporal region. Following initiation of antiepileptic treatment, his neurological examination normalized concomitantly with resolution of the NCSE. A follow-up MRI scan of the brain obtained 1 month later was normal. NCSE can have unusual clinical manifestations, and a high index of suspicion is necessary to correctly diagnose these patients. PMID- 17418646 TI - A new framework for identifying combinatorial regulation of transcription factors: a case study of the yeast cell cycle. AB - By integrating heterogeneous functional genomic datasets, we have developed a new framework for detecting combinatorial control of gene expression, which includes estimating transcription factor activities using a singular value decomposition method and reducing high-dimensional input gene space by considering genomic properties of gene clusters. The prediction of cooperative gene regulation is accomplished by either Gaussian Graphical Models or Pairwise Mixed Graphical Models. The proposed framework was tested on yeast cell cycle datasets: (1) 54 known yeast cell cycle genes with 9 cell cycle regulators and (2) 676 putative yeast cell cycle genes with 9 cell cycle regulators. The new framework gave promising results on inferring TF-TF and TF-gene interactions. It also revealed several interesting mechanisms such as negatively correlated protein-protein interactions and low affinity protein-DNA interactions that may be important during the yeast cell cycle. The new framework may easily be extended to study other higher eukaryotes. PMID- 17418647 TI - Lung transplantation for cystic fibrosis: current concepts and one center's experience. AB - BACKGROUND: Although new approaches to the treatment of patients with cystic fibrosis (CF) are significantly prolonging their lives, most patients will eventually develop respiratory failure due to progressive bronchiectasis caused by chronic lung infection and inflammation and die from to respiratory failure. We examined our center's (University of Wisconsin Hospital and Clinics) experience with lung transplantation for patients with CF and reviewed the literature to examine current and evolving approaches to transplantation for this indication. METHODS: We reviewed all published literature pertaining to lung transplantation for CF through 2006, and we reviewed all aspects of transplantation for patients with CF at our institution from 1994 to 2005. RESULTS: Major complications following lung transplantation include acute rejection, bacterial infection, and bronchiolitis obliterans. Five-year survival at UWHC (Kaplan-Meier) is 67%, and survival was not adversely affected by transplanting patients receiving mechanical ventilation. The major cause of death for transplant recipients was bronchiolitis obliterans syndrome (BOS). CONCLUSIONS: Lung transplantation for CF is associated with acceptable survival rates and can improve quality of life. Lung transplant should be offered to all patients with advanced CF lung disease if they meet currently accepted inclusion and exclusion criteria. PMID- 17418648 TI - Control of electroosmotic flow by a cation additive to enhance the separation of amino acids by micellar electrokinetic chromatography. AB - The effect of a divalent cation (Mg2+) and 3 monovalent cations (Na+, Li+, and K+) as buffer additives on the electroosmotic flow (EOF) was investigated in order to improve the separation performance of p-acetamidobenzenesulfonyl fluoride (PAABS-F) derivatives of 20 standard amino acids by micellar electrokinetic chromatography (MEKC). The EOF can be decreased with increasing concentration of cations with the order of cations as Mg2+>K+>Na+>Li+. However, it was found that the resolution cannot be improved easily using a buffer cation which is more capable of decreasing the EOF. Taking the migration time, resolution, and peak shape into account, Na+ is the best buffer additive, although the EOF decreased most using Mg2+. Using 20 mmol/L borate at pH 9.3 containing 140 mmol/L SDS and 20 mmol/L Na+ as electrolyte, 20 standard amino acids were successfully separated within 14 min. PMID- 17418649 TI - Breath air analysis and its use as a biomarker in biological monitoring of occupational and environmental exposure to chemical agents. AB - The analysis of exhaled air has several advantages since it is a noninvasive method applicable to a large number of toxic agents, in addition to being a simpler matrix than those of other biological samples such as urine and blood. However, it presents some challenges, such as the necessity of a more sensitive sampling procedure, since the chemical substances eliminated through exhaled air are unchanged in form, not being metabolized, and exhaled compounds are present at extremely low concentrations, i.e. in the nanomolar range. To improve the sensitivity and precision of measurement of the concentration of these substances in exhaled air, the sample usually has to be concentrated before assay by gas chromatography. To this end, the use of the solid-phase microextraction (SPME) technique has been proposed as an efficient sampling method. This paper presents a revision of breath analysis as a biomarker for occupational and environmental exposure to chemicals. The sampling methods and the potential use of SPME for determining chemical substances in exhaled air are discussed. PMID- 17418650 TI - Determination of ascorbic acid and its related compounds in foods and beverages by hydrophilic interaction liquid chromatography. AB - A new hydrophilic interaction liquid chromatography method for the simultaneous determination of ascorbic acid (AA), erythorbic acid (EA), 2-O-alpha-D glucopyranosyl-L-ascorbic acid (AA-2G) and 2-O-beta-D-glucopyranosyl-L-ascorbic acid (AA-2betaG) was developed using a diol column with an isocratic solution of acetonitrile-66.7 mM ammonium acetate solution (85:15, v/v) at a detection wavelength of 260 nm. The calibration curves were found to be linear in the range of 1-50 microg/ml for AA and EA and in the range of 2.5-100 microg/ml for AA-2G and AA-2betaG. Detection limits of AA, EA, AA-2G and AA-2betaG were 0.3, 0.3, 0.03 and 0.03 microg/ml, respectively. This method was satisfactorily applied to the determination of AA, EA, AA-2G and AA-2betaG in a fruit, a food and beverages. The results show that the procedure is simple and sensitive and that it can be employed for the simultaneous determination of AA and its related compounds in foods and beverages. PMID- 17418652 TI - Perspectives on this issue of the IJS. PMID- 17418651 TI - Synthesis and characterization of segmented poly(esterurethane urea) elastomers for bone tissue engineering. AB - Segmented polyurethanes have been used extensively in implantable medical devices, but their tunable mechanical properties make them attractive for examining the effect of biomaterial modulus on engineered musculoskeletal tissue development. In this study, a family of segmented degradable poly(esterurethane urea)s (PEUURs) were synthesized from 1,4-diisocyanatobutane, a poly(epsilon caprolactone) (PCL) macrodiol soft segment and a tyramine-1,4-diisocyanatobutane tyramine chain extender. By systematically increasing the PCL macrodiol molecular weight from 1100 to 2700Da, the storage modulus, crystallinity and melting point of the PCL segment were systematically varied. In particular, the melting temperature, T(m), increased from 21 to 61 degrees C and the storage modulus at 37 degrees C increased from 52 to 278MPa with increasing PCL macrodiol molecular weight, suggesting that the crystallinity of the PCL macrodiol contributed significantly to the mechanical properties of the polymers. Bone marrow stromal cells were cultured on rigid polymer films under osteogenic conditions for up to 21 days. Cell density, alkaline phosphatase activity, and osteopontin and osteocalcin expression were similar among PEUURs and comparable to poly(d,l lactic-coglycolic acid). This study demonstrates the suitability of this family of PEUURs for tissue engineering applications, and establishes a foundation for determining the effect of biomaterial modulus on bone tissue development. PMID- 17418653 TI - Anti-inflammatory and immunomodulatory strategies to protect the perinatal brain. AB - Infection and inflammation contribute to perinatal brain damage, particularly to the white matter. Although combating perinatal inflammation can be dangerous, because inflammation might have beneficial effects for mother and fetus, it is worthwhile reviewing potential anti-inflammatory neuroprotective compounds, along with their potential adverse effects. Further research on the possible neuroprotective roles of existing medications and substances is necessary. PMID- 17418654 TI - Distal phalanx replantation using the delayed venous method: a high success rate in 21 cases without specialised technique. AB - BACKGROUND: The purpose of the study was to show that the delayed venous method provides a high success rate in distal phalanx replantation, and does not require the use of specialised techniques. Vein anastomosis is the most important factor determining the 'take rate' in treatment of distal phalanx amputation. However, blood flow in the distal phalanx subdermal vein is lost immediately after an accident, making it difficult to find the collapsed vein and to perform vein anastomosis in the initial surgery. Therefore, we have chosen to perform a two stage surgical procedure, and we have obtained excellent results with this method. METHODS: The two-stage delayed venous method for vein anastomosis was first reported in 2005. This surgical procedure includes initial arterial anastomosis, delayed expansion of the vein, and subsequent vein anastomosis in a second surgery. RESULTS: The delayed venous method was used in 21 cases. Expansion of veins of up to 1mm or more resulted in a high success rate (85.7%) in procedures performed by a junior microsurgeon. In contrast, the success rate for distal phalanx replantation is extremely low in other techniques because of difficulty with vein anastomosis. CONCLUSIONS: The delayed venous method allows relatively easy anastomosis of the subdermal vein of the distal phalanx. Furthermore, the procedure was performed by a junior microsurgeon with less than 2 years experience, showing that the method does not require special training. Therefore, it is a useful operative technique for treatment of amputated fingers by a non-specialised plastic surgeon. PMID- 17418655 TI - Effects of clopidogrel on survival of rat skin flaps. AB - Clopidogrel is a thienopyridine derivative that is chemically related to ticlopidine, which irreversibly inhibits platelet aggregation by selectively binding to adenylate cyclase-coupled adenosine diphosphate receptors on the platelet's surface. In animal models, clopidogrel has been shown to reduce the incidence of both arterial and venous thrombi. In the present study the effects of clopidogrel on the survival of rat epigastric island flaps was researched. Epigastric island flaps of 7x7cm were raised from symphisis pubis to arcus costa with the panniculus carnosus. The experimental group received seven doses of 25mg/kg clopidogrel postoperatively, the first dose given immediately after the suturing of the flaps. The rats were anaesthetised on postoperative day 7 to assess the survival of flaps. The difference between the clopidogrel and the control group was significant (P<0.005). The full-thickness skin samples obtained after the calculation of survival percentages revealed thinning of the epidermis layer and active chronic inflammation in both groups. However, diffuse dilated vessels, extravasated eritrocytes were seen in the clopidogrel group flaps. The results indicated a significant increase in flap survival in rats given clopidogrel. Further research is needed to assess the critical doses of clopidogrel to create optimal flap survival improvement. PMID- 17418656 TI - Research priorities of NSW midwives. AB - PURPOSE: Research is vital to achieve optimum health outcomes for pregnant women, children and families. Recently, the benefit of setting research priorities to effectively utilize limited resources has been identified. Currently there is a lack of published Australian research data relevant to the practice of midwifery. Consultation with current practitioners is important to fulfill the National Health and Medical Research Council (NHMRC) key priority for a healthy start to life, ensure limited resources are expended appropriately and promote evidence based midwifery practice. The aim of this study was to ascertain the perceived research priorities and the research experience of midwives in NSW, Australia. PROCEDURES: Postal questionnaire sent to the 1000 subscribers of Australian Midwifery, of whom 90% (900) are midwives, in March 2005 with key open-ended questions to ascertain midwifery research priorities and research experience of participants. FINDINGS: Respondents were all midwives with 95% indicating they were currently practising as a midwife. They identified six priority areas: professional practice; clinical issues; education and support; breastfeeding; psychosocial factors; rural/indigenous issues. PRINCIPLE CONCLUSIONS: Priorities for research were identified and the need for a link between research and professional midwifery practice was highlighted. Midwives were positive about the possibility of becoming more actively involved in research and/or advocates for evidence based practice. The opportunity exists to take the broad priority areas from this study and develop research questions of relevance for the midwifery profession. PMID- 17418657 TI - Marine Pollution Bulletin annual prize. PMID- 17418659 TI - Tradition and innovation: finding the right balance. PMID- 17418660 TI - The status of US allergy/immunology physicians in the 21st century: a report from the American Academy of Allergy, Asthma & Immunology Workforce Committee. AB - The American Academy of Allergy, Asthma & Immunology has tracked the US allergy/immunology physician workforce (AIPW) over the past 3 decades by funding 2 workforce surveys (1999, 2004). Results have demonstrated both accomplishments of and challenges for the US AIPW. Accomplishments include increases in diversity (25% women in 2004, 20% in 1999, 10% in 1989; 6% underrepresented minorities in 2004, 5% in 1999), 95% of AIPW has completed an allergy/immunology (A/I) training program, and 91% are American Board of Allergy and Immunology (a conjoint board of the American Board of Internal Medicine and the American Board of Pediatrics) certified (90% in 1999). Training positions and program numbers are slowly increasing, and numbers of new graduates from accredited A/I programs have also increased. We are seeing patients with more complex allergic and immune diseases and giving less allergen immunotherapy. Personal, professional, and economic satisfaction is increasing. Challenges relate primarily to diminishing practitioner supply (4245 in 2004 vs 4356 in 1999) amid growing US population demand. The AIPW is gradually aging (the average age is 53 years in 2004, compared with 51 years in 1999) and working longer before retiring. The combination of job satisfaction, the high demand for A/I services, and the large number of fellowship applicants all support expanding the supply of trained allergists/immunologists. PMID- 17418661 TI - Allergic rhinitis and its impact on asthma update: allergen immunotherapy. AB - The Allergic Rhinitis and its Impact on Asthma document was first published in 2001. Since then, new data on specific immunotherapy have appeared. This review is intended as an update to the original document. MedLine (2001 to June 2006) was searched with appropriate key words, and panelists were asked to identify further relevant articles. Randomized controlled trials were considered for the evaluation of efficacy. For the evaluation of safety and additional effects, studies with lower grades of evidence were included. The clinical efficacy of injection immunotherapy in rhinitis and asthma was confirmed, as well as the safety, provided that recommendations are followed. Studies have demonstrated the long-term efficacy and the preventive effect of immunotherapy in reducing the onset of new sensitizations. One randomized open trial demonstrated that in children with allergic rhinitis, injection immunotherapy may reduce the risk of developing asthma. There is strong evidence that sublingual immunotherapy is effective in allergic rhinitis in adults. Recent meta-analyses demonstrated its efficacy in allergic rhinitis in children and in asthma, although more definitive trials are required. Current data indicate that sublingual immunotherapy is safe and the rate of adverse reactions is not greater below 5 years of age. One randomized open trial showed that in children with allergic rhinitis, sublingual immunotherapy reduced the onset of asthma. Further studies are needed to identify the optimal maintenance dose and to elucidate the mechanism of action. Novel approaches for immunotherapy are currently under evaluation, including the use of adjuvants, peptides, and DNA-conjugated and recombinant allergens. PMID- 17418662 TI - Update on the current status of peptide immunotherapy. AB - The use of synthetic peptide fragments of allergen molecules holds promise for the delivery of effective immunotherapy without IgE-mediated adverse events. Early studies were associated with frequent induction of allergic symptoms after treatment, mostly related to activation of allergen-specific effector T cells with high doses of peptides. More recently, low doses of peptides have been shown to modify clinical and laboratory surrogates without inducing adverse events. Studies are ongoing to define the optimal dose, dose interval, and route of administration. Current results indicate that treatment with peptides modulates the immune response by reducing T(H)2 responses to allergen and increasing IL-10 production and the activity of allergen-specific regulatory T cells. Further studies are required in larger numbers of subjects and with peptides derived from a variety of allergens. PMID- 17418663 TI - 2006 American Academy of Allergy, Asthma & Immunology member immunotherapy practice patterns and concerns. PMID- 17418664 TI - Philip S Norman and Lawrence M Lichtenstein: the science of immunotherapy. PMID- 17418665 TI - Legends in allergy: Philip S. Norman and Lawrence M. Lichtenstein--the Hopkins experience. PMID- 17418668 TI - Solutions to challenging digital replantations. AB - Replantation of amputated digits remains one of the most challenging areas in plastic and reconstructive surgery. Complicated cases of digital amputation, such as fingertip amputations, multidigital amputations, and so forth, pose an even greater challenge, requiring extraordinary solutions for successful replantation. The authors present their experience with complicated digital replantations at the University of Medicine and Dentistry of New Jersey-New Jersey Medical School. Cases presented include fingertip replantation, replantation of a finger with impairment of arterial inflow, and two cases of multidigital amputations. In one case of multidigital amputation, heterodigital replantation was performed, and in the other case, a minute skin neurovascular free flap from a nonreplantable finger was used for the reconstruction of another injured finger. Presented cases demonstrate various tools that can be successfully used in the performance of challenging digital replantations. PMID- 17418669 TI - Replantation outcomes. AB - Replantation aims to restore the amputated part to its anatomical site, preserving function and appearance. Outcome depends on factors intrinsic to the patient and to the nature of the injury. Young patients who have distal, cleanly amputated extremities have the best return of function; multiple levels of injury, crush, or avulsing injuries have less. Patients must be fully informed about the commitment to rehabilitation and the possibility of multiple surgeries needed for best results. Similarly, patient and surgeon expectations should be evaluated and addressed before replantation. Meticulous microsurgical technique, comprehensive occupational therapy, and perseverance are needed for success. Addressing these issues promotes a team rehabilitation to restore function while getting the amputation patient back to productive position in society. PMID- 17418670 TI - Ring avulsion: injury pattern, treatment, and outcome. AB - Ring avulsion injury is classically believed to be one of the most challenging for the reconstructive surgeon. Injury patterns include a spectrum from laceration to complete amputation, with differential magnitude of injury to soft tissue, tendon, joint complex, and bone. Although ring avulsion injury can result in devastating functional and aesthetic loss to the hand, fear of even greater functional loss has made replantation a relative contraindication for this complex injury. This article hopes to clarify the pathomechanics and anatomy of ring avulsion, such that the elements of injury, which lead to poor survival and poor function, can be adequately addressed in the surgical plan. PMID- 17418671 TI - Nonextremity replantation: the management of amputations of the facial parts and testicle. AB - Successful nonextremity replantations, particularly of the facial anatomy and testicles, are rare procedures, and only a handful of cases have been reported. This article reviews the current literature in nonextremity replantations and representative cases performed at the authors' institution. Certain underlying themes and problems are consistently encountered in the surgical management of these cases. These are identified and reviewed. Although the replantation of these body parts remains technically challenging, all efforts should be made, when indicated, to repair these injuries microsurgically, because it currently offers the best reconstructive solution for these patients. PMID- 17418672 TI - Extremity reconstruction using nonreplantable tissue ("spare parts"). AB - After a severe digital or extremity injury, the replantation surgeon should always seek to make the best use out of what tissue is available for reconstruction. Exercising sound surgical judgment and being creative allow the surgeon to restore function to critical areas of the hand or extremity by the judicious use of available tissues that would otherwise be discarded. The use of "spare parts" should, therefore, always be considered to facilitate digital or extremity reconstruction when routine replantation is not possible or is likely to produce a poor functional result. The surgeon should always try to use available nonreplantable tissue to preserve length, obtain soft tissue coverage, or most importantly improve the function of remaining less injured digits. This article presents several case studies that illustrate the principals of spare parts reconstruction performed at the time of the initial debridement using nonreplantable tissue to provide coverage or improve function. PMID- 17418673 TI - Great toe-to-thumb microvascular transplantation. AB - Great toe to thumb transplantation offers the ideal reconstruction because it replaces the absent thumb with nearly identical anatomy. Functional donor site morbidity is minimal as compared with the aesthetic one; however, in comparison to the hand, the functional and aesthetic gain is tremendous. Anatomic and operative details are discussed along with preoperative and postoperative management. Postoperative functional evaluation of the thumb and donor site is assessed. Forty years of experience with this composite tissue transplantation has proven its status as the gold standard for thumb reconstruction. PMID- 17418674 TI - Research and events leading to facial transplantation. AB - Facial transplantation has long captured the interest and imagination of scientists, the media, and the lay public. Facial transplantation could provide an excellent alternative to current treatments for facial disfigurement caused by burns, trauma, cancer extirpation, or congenital birth defects. This article discusses the major technical, immunologic, psychosocial and ethical hurdles that have been overcome to bring facial transplantation from an idea to a clinical reality by providing the reader with a chronologic overview of the research and events that have led this exciting new treatment into the clinical arena. PMID- 17418675 TI - Tissue transplantation in plastic surgery. AB - The functional and aesthetic outcome following application of conventional reconstructive procedures or prosthetic materials is not satisfactory, especially in patients who have severe deformities and disabilities. Since the first successful hand transplantation in France in 1998, composite tissue allograft transplantation has gained a great deal of interest in the field of plastic surgery. It is obvious that composite tissue allograft transplantation will improve patients' life quality, but this might be at the expense of decreasing life expectancy. Currently, the main obstacle for composite tissue allograft transplantation is the use of life-long immunosuppression therapy because of their well-known side effects. In addition, the ethical, social, and psychologic issues are raised when discussing face transplantation. The long-term results of the recently performed partial face transplantations will be critical to judge the future applications of partial or total face transplantation. PMID- 17418676 TI - Transplantation of the hand, face, and composite structures: evolution and current status. AB - This article reviews the world experience in the newly emerging field of composite tissue allotransplantation. These allografts contain multiple tissues that are usually musculoskeletal structures with a skin or epithelial surface, such as hand, facial structures, larynx, tongue, ear, knee/femur, abdominal wall, and penis. They represent a new transplantation field, with only a 10-year experience and just over 50 clinical cases. This review of the 10-year world experience found uniform technical success, immunologic biology, and immunosuppression regimens very similar to solid organ transplants, and success strongly correlated with adherence to guidelines for psychiatric screening, thorough preparation of patient and families, intense postoperative monitoring, and assurance of medication access. All failures reported have been caused by lapses in these parameters. This early experience shows a great potential for application of these new procedures to the most challenging reconstructive needs. PMID- 17418677 TI - Creating a hand transplant program. AB - Successful hand transplantation depends upon an extensive, well-coordinated system with resources and experience in both upper extremity and transplant surgery. This description of such a system is drawn from the experience of the Louisville, Kentucky, Hand Transplant Program, which achieved the world's first successful hand transplant in January 1999, and which is the only United States program to date. The personnel and components of this system are outlined, with a detailed description of the roles, responsibilities, and interaction between components. This system ensures optimal management throughout recipient selection, psychiatric screening, education and preparation, donor limb retrieval, transplant surgery, immunosuppression administration and monitoring, and post-transplant rehabilitation therapy. The lessons learned from the experience of the Louisville team, and the experiences of the other teams world wide, are presented to guide organization of future teams participating in this pioneering endeavor. PMID- 17418678 TI - Cellular therapies for prolongation of composite tissue allograft transplantation. AB - Complex musculoskeletal defects resulting from cancer, congenital absence, and trauma represent a unique reconstructive challenge. Autologous tissue is often unavailable to reconstruct these deformities. Composite tissue allograft transplantation represents a unique solution for these clinical problems. Face, hand, or limb transplants can be performed in a single procedure. However, the use of chronic nonspecific systemic immunosuppression can lead to side effects such as drug toxicity, opportunistic infections, and malignancies. This article explores various cell-based therapies that represent promising modalities to reduce chronic immunosuppression and alter the risk/benefit ratios for the prospect of composite tissue allograft transplantation. PMID- 17418679 TI - The immunology of composite tissue transplantation. AB - Composite tissue allotransplantation holds great potential for reconstructive surgery. That these procedures can be successful has been clearly demonstrated by the success of hand, face, and larynx transplants around the world. Although the immunology of composite tissue allotransplantation mirrors that of any allogeneic organ transplant, there are several unique aspects to these grafts. This article reviews the immunology of transplantation, histocompatibility testing for composite tissue allotransplantation, graft rejection, immunosuppression, and specific immunologic considerations of composite tissue allotransplantation. PMID- 17418680 TI - Tissue engineering: bridging the gap between replantation and composite tissue allografts. AB - This article explores issues related to tissue engineering and composite tissue allografts that employ physiologic and anatomic autogenous replicates to restore tissue loss. Composite tissue allotransplantation has become a controversial option for reconstruction, most prominently for reconstruction involving the hand and, recently, the face. While the side-effect profile of systemic immunosuppression continues to improve, the long-term risks of immunosuppression leaves composite tissue allotransplantation a domain for cautious exploration. Meanwhile, tissue engineering could, conceivably, be the gap between replantation and composite tissue allografts. Whereas the perils of immunosuppression may limit the routine use of allografts, employing constructions made of the patient's own cells negates the need for any antirejection therapy. PMID- 17418681 TI - Diabetic nephropathy: introduction. PMID- 17418682 TI - AGE, RAGE, and ROS in diabetic nephropathy. AB - Diabetic nephropathy is a major cause of morbidity and mortality in diabetic patients. Two key mechanisms implicated in the development of diabetic nephropathy include advanced glycation and oxidative stress. Advanced glycation is the irreversible attachment of reducing sugars onto amino groups of proteins to form advanced glycation end products (AGEs). AGE modification of proteins may lead to alterations in normal function by inducing cross-linking of extracellular matrices. Intracellular formation of AGEs also can cause generalized cellular dysfunction. Furthermore, AGEs can mediate their effects via specific receptors, such as the receptor for AGE (RAGE), activating diverse signal transduction cascades and downstream pathways, including generation of reactive oxygen species (ROS). Oxidative stress occurs as a result of the imbalance between ROS production and antioxidant defenses. Sources of ROS include the mitochondria, auto-oxidation of glucose, and enzymatic pathways including nicotinamide adenine dinucleotide phosphate reduced (NAD[P]H) oxidase. Beyond the current treatments to treat diabetic complications such as the optimization of blood pressure and glycemic control, it is predicted that new therapies designed to target AGEs, including AGE formation inhibitors and cross-link breakers, as well as targeting ROS using novel highly specific antioxidants, will become part of the treatment regimen for diabetic renal disease. PMID- 17418683 TI - The renin-angiotensin system and diabetic nephropathy. AB - The renin-angiotensin system (RAS) has key regulatory functions for blood pressure and fluid homeostasis. In addition, dysregulation of the system can have maladaptive effects to promote tissue injury in chronic diseases such as hypertension, heart failure, and kidney disease. These actions for the RAS to promote disease pathogenesis are especially apparent in diabetic nephropathy, the most common cause of end-stage renal disease in the United States. Evidence of a role for the RAS in diabetic nephropathy comes from studies in animal models and randomized clinical trials showing efficacy of angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers to slow the progression of renal disease. Widespread applications of these therapies to a range of renal diseases may have contributed to the recent reduction in the incidence rates for end-stage renal disease. We provide a general review of the RAS and its role in diabetic nephropathy. PMID- 17418685 TI - Angiogenesis in diabetic nephropathy. AB - Angiogenesis, the formation of new blood vessels from pre-existing vasculature, plays a key role in both physiologic and pathologic events, including wound healing, cancer, and diabetes. Neovascularization has been implicated in the genesis of diverse diabetic complications such as retinopathy, impaired wound healing, neuropathy, and, most recently, diabetic nephropathy. Diabetic nephropathy is one of the major microvascular-associated complications in diabetes and is the leading cause of end-stage renal disease worldwide. In this review we describe the major factors involved in the pathologic glomerular microvascular alterations in response to hyperglycemia and the possible use of anti-angiogenic therapies for the treatment of diabetic nephropathy. PMID- 17418684 TI - Regulation of transforming growth factor beta in diabetic nephropathy: implications for treatment. AB - The recognition that the drivers of matrix accumulation is an appropriate therapeutic target for diabetic nephropathy is now accepted by the nephrology and pharmaceutical communities. Interventions focused around transforming growth factor-beta (TGF-beta) likely will be an important area of clinical investigation in the near future. Understanding the various pathways involved in stimulating TGF-beta in the diabetic kidney is of paramount importance in devising strategies to combat the development and progression of diabetic nephropathy. In this review we highlight the major pathways involved in stimulating TGF-beta production by increased glucose levels and discuss the therapeutic implications thereof. PMID- 17418686 TI - Albuminuria: a target for treatment of type 2 diabetic nephropathy. AB - Both renal and cardiovascular morbidity and mortality is increased markedly in patients with type 2 diabetes. Besides the classic risk factors and markers such as glucose, blood pressure, blood lipid profile, and lifestyle (smoking, overweight), novel risk markers are identified, among them urine albumin excretion. Levels of urinary albumin excretion greater than normal are observed frequently in patients with type 2 diabetes. Moderately increased levels of albuminuria, so-called microalbuminuria, are predictive both for progressive renal function loss to diabetic nephropathy, and for cardiovascular morbidity and mortality: the higher the albuminuria level, the more chance of renal and cardiovascular complications. More advanced levels of albuminuria (overt albuminuria) are observed in patients in the diabetic nephropathy state. In this condition, renal and cardiovascular risk are extremely high, and again one may observe that the level of albumin excretion is predictive of renal and cardiovascular outcome. Several drug strategies decrease the level of urinary albumin excretion in type 2 diabetic patients. Data on using drugs that intervene in the renin-angiotensin-aldosterone-system (RAAS) are the most extensive and conclusive. RAAS intervention is a very effective strategy to decrease the amount of albumin in the urine, independent from the blood pressure decreasing characteristics of the treatment. RAAS intervention is associated with long-term renal and cardiovascular protection. Importantly, the degree of short-term albuminuria decrease is associated with the degree of renal and cardiovascular protection: the more albuminuria reduction, the more protection. The protective predictive power of the albuminuria effect of RAAS intervention is not related to (or dissociated from) the blood pressure decreasing effect of these drugs. The protective effect of RAAS intervention is present at normoalbuminuric, microalbuminuric, and overt albuminuria levels. This makes albuminuria a target for therapy in type 2 diabetes. New drug strategies that decrease or prevent albuminuria without affecting other risk factors currently are being tested, and not only will add to underscoring the need to treat albuminuria as a separate target, but also will assist in reducing the enormous residual risk burden of individual diabetic patients. PMID- 17418687 TI - Treating hypertension in the patient with overt diabetic nephropathy. AB - Arterial blood pressure is a major determinant of renal and cardiovascular outcomes in diabetic nephropathy. There is a proportional relationship between the systolic blood pressure and renal and mortality outcomes. Decreasing the diastolic pressure does not significantly decrease these outcomes. Irrespective of the magnitude of pretreatment systolic hypertension in the patient with type 2 diabetic nephropathy, the systolic pressure achieved with antihypertensive therapy is the important determinant of renal and cardiovascular risk. Achieving a lower systolic pressure down to 120 mm Hg is associated with substantial risk reduction. Although the data are limited, systolic blood pressure less than 120 mm Hg may be associated with increased all-cause mortality in this patient population, increasing the possibility of a J-curve response. A marked decrease in diastolic pressure, which is a danger when undertaking aggressive therapy with the goal of decreasing the systolic pressure to 130 mm Hg, can be associated with an increased risk of cardiac events. The renoprotective and proteinuria decreasing effects of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers recommend these agents as the standard of care in type 2 diabetic nephropathy. In addition to angiotensin-converting enzyme inhibitor and angiotensin-receptor blocker therapy, controlling the systolic blood pressure in this difficult to control patient population may require the use of 3 or more antihypertensive agents. PMID- 17418688 TI - Histopathology of diabetic nephropathy. AB - The clinical manifestations of diabetic nephropathy, proteinuria, increased blood pressure, and decreased glomerular filtration rate, are similar in type 1 and type 2 diabetes; however, the renal lesions underlying renal dysfunction in the 2 conditions may differ. Indeed, although tubular, interstitial, and arteriolar lesions are ultimately present in type 1 diabetes, as the disease progresses, the most important structural changes involve the glomerulus. In contrast, a substantial subset of type 2 diabetic patients, despite the presence of microalbuminuria or proteinuria, have normal glomerular structure with or without tubulointerstitial and/or arteriolar abnormalities. The clinical manifestations of diabetic nephropathy are strongly related with the structural changes, especially with the degree of mesangial expansion in both type 1 and type 2 diabetes. However, several other important structural changes are involved. Previous studies, using light and electron microscopic morphometric analysis, have described the renal structural changes and the structural-functional relationships of diabetic nephropathy. This review focuses on these topics, emphasizing the contribution of research kidney biopsy studies to the understanding of the pathogenesis of diabetic nephropathy and the identification of patients with a higher risk of progression to end-stage renal disease. Finally, evidence is presented that the reversal of established lesions of diabetic nephropathy is possible. PMID- 17418689 TI - Mining the genome for susceptibility to diabetic nephropathy: the role of large scale studies and consortia. AB - Approximately 30% of individuals with type 1 and type 2 diabetes develop persistent albuminuria, lose renal function, and are at increased risk for cardiovascular and other microvascular complications. Diabetes and kidney diseases rank within the top 10 causes of death in Westernized countries and cause significant morbidity. Given these observations, genetic, genomic, and proteomic investigations have been initiated to better define basic mechanisms for disease initiation and progression, to identify individuals at risk for diabetic complications, and to develop more efficacious therapies. In this review we have focused on linkage analyses of candidate genes or chromosomal regions, or coarse genome-wide scans, which have mapped either categorical (chronic kidney disease or end-stage renal disease) or quantitative kidney traits (albuminuria/proteinuria or glomerular filtration rate). Most loci identified to date have not been replicated, however, several linked chromosomal regions are concordant between independent samples, suggesting the presence of a diabetic nephropathy gene. Two genes, carnosinase (CNDP1) on 18q, and engulfment and cell motility 1 (ELMO1) on 7p14, have been identified as diabetic nephropathy susceptibility genes, but these results require authentication. The availability of patient data sets with large sample sizes, improvements in informatics, genotyping technology, and statistical methodologies should accelerate the discovery of valid diabetic nephropathy susceptibility genes. PMID- 17418690 TI - The application of the HapMap to diabetic nephropathy and other causes of chronic renal failure. AB - The human nuclear genome consists of approximately 3 billion nucleotides. Human beings are 99% similar in DNA sequence to each other, but natural genetic variation in approximately 1% of the DNA sequence is responsible for interindividual differences, including determining who will develop disease and who will remain healthy. The pace and timing of disease initiation also is regulated by exposure to individual-level environmental factors and other random causes. Therefore, an examination of the DNA sequences of individuals with and without diabetic nephropathy, or, more broadly, chronic renal failure, can predict which sequence differences vary with disease (or health). The technology is not yet economical enough to analyze large numbers of individuals down to each nucleotide, but standardized dense genotyping sets for interrogating 1 marker for every 5,000, 10,000, or 15,000 nucleotides now are affordable even in large samples. The swiftness with which disease-gene associations can be mined has improved radically as a result of the availability of discovery human genetic variation data from large-scale public and private initiatives, such as those provided by the International Haplotype Map Consortium and Perlegen Sciences, Inc. (Mountain View, CA). These projects have captured many of the common genetic variants (>1%) in the genome. This information has been buttressed with improvements in large-scale genotyping technologies and statistical methods for data analysis. In summary, the renal community is now poised for discovery of genes for chronic renal failure using these resources. PMID- 17418691 TI - Genetics of diabetic nephropathy: lessons from mice. AB - Although diabetic nephropathy occurs only in a minority of diabetic patients (approximately 30%), it is the major single cause of end-stage renal disease in the United States. Hyperglycemia and hypertension are important factors predisposing patients to nephropathy, however, accumulating evidence points to critical genetic factors that predispose only a subset of diabetic patients to nephropathy. Defining the genes responsible for nephropathy risk in human populations has proven challenging. Comparative genomics using the robust genetic reagents available in the laboratory mouse should provide a complementary approach to defining genes that may predispose to diabetic nephropathy in mice and human beings. In this article we review studies that have started to identify genetic risk factors for diabetic nephropathy in mice and the multiple approaches that may be used to elucidate the genetic pathogenesis of this disorder. PMID- 17418693 TI - Delta receptor antagonism, ethanol taste reactivity, and ethanol consumption in outbred male rats. AB - Naltrexone, a nonspecific opioid antagonist, produces significant changes in ethanol responsivity in rats by rendering the taste of ethanol aversive as well as producing a decrease in voluntary ethanol consumption. The present study investigated the effect of naltrindole, a specific antagonist of delta opioid receptors, on ethanol taste reactivity and ethanol consumption in outbred rats. In the first experiment, rats received acute treatment of naltrexone, naltrindole, or saline followed by the measurement of ethanol consumption in a short-term access period. The second experiment involved the same treatments and investigated ethanol palatability (using the taste-reactivity test) as well as ethanol consumption. Results indicated that treatment with 3 mg/kg naltrexone significantly affected palatability (rendered ethanol more aversive, Experiment 2) and decreased voluntary ethanol consumption (Experiments 1 and 2). The effects of naltrindole were inconsistent. In Experiment 1, 8 mg/kg naltrindole significantly decreased voluntary ethanol consumption but this was not replicated in Experiment 2. The 8 mg/kg dose produced a significant increase in aversive responding (Experiment 2) but did not affect ingestive responding. Lower doses of naltrindole (2 and 4 mg/kg) were ineffective in altering rats' taste-reactivity response to and consumption of ethanol. While these data suggest that delta receptors are involved in rats' taste-reactivity response to ethanol and rats' ethanol consumption, it is likely that multiple opioid receptors mediate both behavioral responses. PMID- 17418694 TI - Ethanol blocks nicotine-induced seizures in mice: comparison with midazolam and baclofen. AB - Low doses of ethanol may antagonize the pharmacological effects of nicotine. Recently, it has been shown that the effects of ethanol on nicotine discrimination are not correlated with blood ethanol levels. The aim of the present study was to evaluate whether ethanol (0.5-2g/kg, i.p.) could block nicotine-induced seizures in C57BL/6J mice and to correlate ethanol's actions with blood ethanol concentrations. For comparison, the effects of a gamma aminobutyric acid A (GABAA)/benzodiazepine receptor positive modulator, midazolam (0.25-40 mg/kg, i.p.), and a gamma-aminobutyric acid B receptor agonist, baclofen (2.5-20 mg/kg, i.p.), were assessed in the same procedure. Nicotine (3-9 mg/kg, s.c.) induced clonic-tonic seizures in a dose-dependent manner. Ethanol, administered 5 or 50 min before nicotine, dose dependently antagonized seizures elicited by 6 mg/kg nicotine. The anticonvulsant effects of ethanol correlated with blood ethanol levels and were comparable to those exerted by midazolam. Baclofen antagonized only the tonic component of nicotine-induced convulsions. The anticonvulsant doses of ethanol (0.5-2 g/kg), midazolam (0.5-1 mg/kg), and baclofen (5-10 mg/kg) did not affect spontaneous locomotor activity in a control experiment. The present results indicate that (i) ethanol may block nicotine induced seizures in mice at doses that do not alter locomotor activity and (ii) the anti-seizure effects of ethanol depend on blood ethanol levels and are comparable to those exerted by the GABAA positive modulator midazolam. PMID- 17418695 TI - Lifelong ethanol consumption and brain regional GABAA receptor subunit mRNA expression in alcohol-preferring rats. AB - Brain regional gamma-aminobutyric acid type A (GABAA) receptor subunit mRNA expression was studied in ethanol-preferring AA (Alko, Alcohol) rats after moderate ethanol drinking for up to 2 years of age. In situ hybridization with oligonucleotide probes specific for 13 different subunits was used with coronal cryostat sections of the brains. Selective alterations were observed by ethanol exposure and/or aging in signals for several subunits. Most interestingly, the putative highly ethanol-sensitive alpha4 and beta3 subunit mRNAs were significantly decreased in several brain regions. The age-related alterations in alpha4 subunit expression were parallel to those caused by lifelong ethanol drinking, whereas aging had no significant effect on beta3 subunit expression. The results suggest that prolonged ethanol consumption leading to blood concentrations of about 10 mM may downregulate the mRNA expression of selected GABAA receptor subunits and that aging might have partly similar effects. PMID- 17418696 TI - Activation of serotonin-3 receptors increases dopamine release within the ventral tegmental area of Wistar and alcohol-preferring (P) rats. AB - The objectives of the present study were to (a) examine the effects of activating serotonin-3 (5-HT3) receptors on dopamine (DA) release in the anterior and posterior ventral tegmental area (VTA) of Wistar rats and (b) determine if there are differences in 5-HT3--stimulated DA release in the VTA between alcohol preferring (P) and Wistar rats. Local perfusion with the 5-HT3 agonist 1-(m chlorophenyl)-biguanide (CPBG) in the anterior and posterior VTA stimulated DA release in both the regions. The CPBG-stimulated increase in extracellular DA levels was significantly higher in the posterior than anterior VTA of Wistar rats. The basal extracellular DA levels were not significantly different between the anterior and posterior VTA of Wistar rats. However, the basal extracellular DA levels were significantly higher in the posterior VTA of Wistar rats than P rats. Local perfusion of CPBG into the posterior VTA stimulated somatodendritic DA release significantly more in the P than Wistar rat. Overall, the results indicate that there may be a heterogeneous distribution of functional 5-HT3 receptors within the VTA, with higher numbers in the posterior than anterior VTA, and that, compared to 5-HT3 receptors in Wistar rats, 5-HT3 receptors in the posterior VTA of P rats may be more responsive to stimulation. PMID- 17418697 TI - Nucleus accumbens serotonin transporters in alcoholics measured by whole hemisphere autoradiography. AB - Nucleus accumbens (NAC) is regulated by the dopaminergic and serotonergic pathways, and it is a brain area with a crucial role in the rewarding effects of ethanol. In this preliminary study, possible alterations of [3H]citalopram binding to serotonin transporter (SERT) were evaluated in the NAC of Cloninger type 1 and 2 alcoholics (nine and seven subjects, respectively), and nonalcoholic controls (10 subjects) by human postmortem whole-hemisphere autoradiography. The [3H]citalopram binding in the NAC was 35% higher in the alcoholics than in the controls; in the type 1 alcoholics, the binding was 54% and in the type 2 alcoholics it was 17% higher. Although the effect size showed medium effects (0.49-0.60), the results did not reach statistical significance due to large standard deviations. The [3H]citalopram binding declined significantly with age in the controls, but not in the alcoholics. In the controls, there was a significant positive correlation between the [3H]citalopram binding in the NAC and in the anterior cingulate gyrus, an area in which the [3H]citalopram binding has been shown to be lower among alcoholics. On the contrary, a significant negative correlation was observed in the type 2 alcoholics and no correlation in the type 1 alcoholics. In addition, there was a strong tendency toward a positive correlation between the SERT and dopamine transporter binding in the type 2 alcoholics, but not in the other groups. These preliminary results suggest a differential monoaminergic imbalance in type 1 and 2 alcoholism in brain areas important for the regulation of motivation, reward, and reinforcement. PMID- 17418698 TI - Combined effects of high-fat diet and ethanol induce oxidative stress in rat liver. AB - Individuals affected by liver steatosis seldom have symptoms of liver injury, but may be particularly vulnerable to oxidative insults. In this study, we evaluated liver redox alterations produced by acute ethanol administration to rats that were fed a high-fat diet (HFD). Adult male Wistar rats were fed HFD or standard diet (controls) for 1 month; a group of animals from each condition were gavaged with 35% (vol/vol) ethanol every 12h for the last 3 days of the experiment. Total lipid content determined in liver showed lipid accumulation after HFD or HFD combined with ethanol. HFD alone induced a significant rise of seric alanine aminotransferase levels and a marked reduction of antioxidant enzyme activities (catalase, superoxide dismutase, glutathione transferase). Ethanol alone caused a significant rise of seric cholesterol levels and enhanced mitochondrial H2O2 production, but without apparent oxidative stress as evaluated by thiobarbituric acid-reactive substances (TBARS) assay. The combination of HFD and acute ethanol caused an increase of TBARS, indicating lipid peroxidation, most likely as a consequence of a decrease in antioxidant defenses induced by HFD and of an increase in reactive oxygen species production induced by ethanol. Principal component analysis, based on all the measured parameters, that is, serum liver function tests, antioxidant enzyme activities, mitochondrial H2O2 release, and TBARS, indicated that HFD and ethanol act as two independent factors. In conclusion, our results show that HFD or acute ethanol alone produce, at the most, mild liver injury, whereas their combination triggers oxidative stress, possibly inducing a progression toward liver disease. Hence, our data indicate that a diet too rich in fat is a serious risk factor for the occurrence of liver injury deriving from acute ethanol consumption. PMID- 17418699 TI - Alcohol-drinking history and fatal injury in older adults. AB - Although most clinical guidelines for older adults allow for one drink a day in persons without a history of alcoholism, diabetes, or cardiovascular disease, alcohol may contribute to fatal injury in the elderly. Using two national surveys, this case-control study determined the associations between drinking history and fatal injuries from falls, motor vehicle crashes and suicides. We performed a case-control study using 1,735 cases who died of falls, motor vehicle crashes, or suicides selected from the 1993 National Mortality Follow-Back Survey; controls (n=13,381) were a representative sample of the U.S. population from the 1992 National Longitudinal Alcohol Epidemiologic Survey. Cases and controls were restricted to ages 55 years and older. Having 12 or more drinks in the year before death or interview for the controls was used to assess alcohol drinking history. The unadjusted relative odds for drinkers versus nondrinkers for falls, motor vehicle crashes, and suicides were 1.7, 1.7, and 1.6, respectively. Adjustment for age, gender, marital status, education, and working in the last year did not change these effect estimates, which all excluded the null value. Drinking increased the risk of suicide more for women than for men. Drinking history in older adults is associated about equally with an increased risk of fatal injury from falls, motor vehicle crashes, and suicides. PMID- 17418702 TI - True molar intrusion. PMID- 17418705 TI - The origin of treatment expansion. PMID- 17418706 TI - RNA interference of Bmp-4 and midface development in postimplantation mouse embryos. AB - INTRODUCTION: Clefts of the lip and palate result when the midfacial processes fail to fuse during embryonic development. Many of the cellular and molecular events underlying the fusion of these midfacial processes are unknown. There is evidence from our laboratory and others that Bmp-4 is a signal that regulates the process of fusion in the midface. RNA interference (RNAi) is a relatively new technique used to knock down select RNA levels--and ultimately protein levels--by using short RNA strands complementary to target mRNA in the cytoplasm. This was the first investigation to use RNAi to study the developing wild-type murine midface in vivo. This study used RNAi technology to decrease levels of Bmp-4 RNA during critical stages of midfacial fusion with the hypothesis that correct temporal and spatial expression of Bmp-4 is critical for normal fusion and formation of the midface. METHODS: The sample included 19 timed pregnant wild type (C57B1/6J strain) mice and their litters. Pregnant mice received tail-vein injections of plasmids coding for an epifluorescent label (DsRed) with or without the Bmp-4 RNAi sequence 3 days before typical embryonic midface fusion. Embryos were harvested before, during, and after the typical date of midface fusion, and delivery of the plasmids was confirmed via epifluoroscopy and reverse transcriptase polymerase chain reaction (RT-PCR) for the DsRed label. RESULTS: Alterations in morphology were observed by gross examination and by sectioning the embryos. Levels of Bmp-4 mRNA and protein in the embryonic midface were examined with RT-PCR and immunohistochemistry, respectively. Injections of RNA plasmids via the maternal tail-vein to pregnant mothers reliably delivered plasmids to the developing embryonic midface with an average 46% (+/-12.2) knockdown in Bmp-4 RNA levels. CONCLUSIONS: Reduced BMP-4 levels caused prefusion delays in the outgrowth of the lateral and medial nasal processes. These processes later recovered, producing no detectable long-term morphological differences when compared with the control embryos. Ectopic Bmp-4 protein expression was observed in the lateral and medial nasal processes mesenchyme in Bmp-4 knockdown embryos superior to the region of primary palatal fusion. These findings implicated Bmp-4 signaling in the outgrowth and development of the midfacial processes. This study introduced the use of RNAi for the study of protein signaling in the developing wild-type murine midface in vivo. PMID- 17418707 TI - Cervical vertebral and dental maturity in Turkish subjects. AB - INTRODUCTION: The aim of this study was to investigate the relationships between the stages of calcification of teeth and the cervical vertebral maturity stages in Turkish subjects. METHODS: A retrospective cross-sectional study was designed. The final study population consisted of 590 Turkish subjects. Statistical analysis of the data was performed with computer software. Spearman rank order correlation coefficients were used to assess the relationship between cervical vertebral and dental maturation. For a better understanding of the relationship between cervical vertebral maturation indexes and dental age, percentage distributions of the studied teeth were also calculated. RESULTS: Strict correlations were found between dental and cervical vertebral maturation of Turkish subjects. For males, the sequence from lowest to the highest was third molar, central incisor, canine, first premolar, second premolar, first molar, and second molar. For females, the sequence from lowest to the highest was third molar, canine, second premolar, first premolar, central incisor, first molar, and second molar. CONCLUSIONS: Dental maturation stages can be used as a reliable indicator of facial growth. PMID- 17418708 TI - Dentoskeletal and soft-tissue changes with cervical headgear and mandibular protraction appliance therapy in the treatment of Class II malocclusions. AB - INTRODUCTION: In this study, we cephalometrically compared changes in the dentoskeletal and soft tissues after orthodontic treatment of Class II Division 1 malocclusions between the cervical headgear (CHG) and the mandibular protraction appliance (MPA) followed by fixed appliances. METHODS: The sample consisted of 50 patients divided into 2 groups of 25 patients each (13 male, 12 female). Group 1 patients were treated with CHG and fixed appliance, and group 2 patients were treated with the MPA and fixed appliances. The patients were matched according to sex, age, treatment time, and initial cephalometric variables. The groups were compared regarding pretreatment stage and treatment changes, with t tests, at P <.05. RESULTS: The CHG group showed statistically greater restriction of the anterior displacement of the maxilla, improvement of the skeletal Class II relationship, decreased facial convexity, extrusion and distalization of the maxillary first molar, and extrusion of the mandibular incisors. The MPA group had statistically greater increases in maxillary length, mandibular protrusion, maxillary incisor retrusion, mesial displacement of the mandibular first molars, and protrusion of the lower lip. CONCLUSIONS: The changes in Class II malocclusion correction between the CHG and the MPA were that the CHG corrected the Class II malocclusion primarily through greater action on the maxillary skeletal and dentoalveolar structures as compared with the MPA, which corrected the malocclusion through greater action on the mandibular dentoalveolar structures. PMID- 17418709 TI - Acceleration of orthodontic tooth movement by alveolar corticotomy in the dog. AB - INTRODUCTION: Tooth movement and alveolar bone reaction after corticotomies have not been thoroughly examined. In this study, the effects of corticotomies on orthodontic tooth movement and alveolar bone reaction were investigated in dogs. METHODS: Corticotomies were performed on the cortical bone of the mandibular left third premolar region in 12 male adult beagles. The third premolars on the left experimental side and on the right sham side were moved mesially with a continuous force of 0.5 N. RESULTS: Tooth movement velocities from 0 to 1 week and from 1 to 2 weeks after the corticotomies were significantly faster on the experimental side than on the sham side. Hyalinization of the periodontal ligament appeared only at 1 week after the corticotomies on the experimental sides, whereas it was observed from 1 to 4 weeks after the corticotomies on the sham sides. Tartrate-resistant-acid-phosphatase positive cells of the experimental side tended to work vigorously at an early time on the alveolar wall and in the bone marrow cavities. CONCLUSIONS: Orthodontic tooth movement increased for at least 2 weeks after the corticotomies. This might be brought about by rapid alveolar bone reaction in the bone marrow cavities, which leads to less hyalinization of the periodontal ligament on the alveolar wall. PMID- 17418710 TI - Periodontal response to early uncovering, autonomous eruption, and orthodontic alignment of palatally impacted maxillary canines. AB - INTRODUCTION: The purpose of this study was to evaluate differences in periodontal status, root length, and visual assessment in patients with palatally impacted maxillary canines that were surgically exposed, allowed to erupt freely into the palate, and orthodontically aligned. METHODS: Clinical examinations of the maxillary lateral incisors, canines, and adjacent premolars were performed on 16 patients with unilaterally impacted canines and 6 with bilaterally impacted canines treated in this manner. The average age was 23 years 7 months, and the average posttreatment observation period was 2 years 11 months. Data from the bilaterally impacted canines were not used in the central analysis. RESULTS: Differences in probing attachment level were found at the distolingual region of the lateral incisor and at the distobuccal region of the premolar adjacent to the treated canine. Crestal bone height was lower at the mesial and distal regions of the lateral incisor adjacent to the previously impacted canine, and the roots of the treated canine and adjacent lateral incisor were shorter than those of the contralateral control teeth. Twenty-three orthodontists and 9 second- and third year orthodontic residents could identify the previously impacted canine in the unilateral patients an average of 78.9% of the time, but to a statistically significant degree in 66% of all patients. CONCLUSIONS: The overall consequences to the impacted canine of surgical exposure and free eruption are good compared with closed exposure and early traction, whereas consequences to the adjacent teeth, particularly the lateral incisor, are similar. Future research directly comparing the 2 methods with a larger sample and randomization could yield further insight. PMID- 17418711 TI - Treatment effects and short-term relapse of maxillomandibular expansion during the early to mid mixed dentition. AB - INTRODUCTION: The treatment effects and the short-term (0.9 +/- 0.45 years) relapse potential of phase I slow maxillary expansion, with a bonded palatal expander or a quad-helix appliance combined with a mandibular banded Crozat/lip bumper and followed by 12 to 15 months of retention, were examined. METHODS: Pretreatment (8.8 +/- 1.7 years) and posttreatment (11.1 +/- 1.7 years) models of 54 patients were used to evaluate treatment effects. Posttreatment (11.0 +/- 1.3 years) and follow-up (11.9 +/- 1.4 years) models of 23 patients who returned for phase II treatment were used to evaluate relapse over the 11 months, during which no retention was used. The models were digitized, and 15 measures were computed. RESULTS: Significant treatment increases were observed for all measurements in both arches. Treatment gains in arch perimeter (6%-8%) were due more to increases in intermolar width (11%-15%) than to increases in arch depth (5%). Posttreatment relapse was significant (P <.05) for all measures except mandibular intercanine width and maxillary molar arch depth. After accounting for normal growth, net changes (pretreatment to follow-up) indicated significant increases for all measures except maxillary molar arch depth. In addition to maintaining leeway space, the maxilla and the mandible showed net perimeter gains of 2.9 and 1.0 mm, respectively. CONCLUSIONS: Slow maxillary expansion combined with a mandibular banded Crozat/lip bumper during the early mixed dentition produced clinically useful increases in arch dimensions that subsequently underwent mild-to-moderate amounts of relapse after removal of all retention appliances. PMID- 17418712 TI - Changes in the masticatory cycle after treatment of posterior crossbite in children aged 4 to 5 years. AB - INTRODUCTION: The aim of this study was to analyze the dynamics of the masticatory cycle after occlusal adjustment through coronoplasty on deciduous teeth (selective grinding) and composite resin occlusal guidance for posterior crossbite correction. METHODS: Nineteen children, aged 4 to 5 years, with functional posterior crossbites diagnosed at their clinical examinations, were selected. Analyses of masticatory cycles, including lateral excursive movements, were performed before and after treatment by using kinesiographic examinations in the 3 reference planes (frontal, horizontal, and sagittal). The examinations were done during bubble-gum chewing. The data were submitted to sign, Mann-Whitney, and paired t tests concerning the variables: left maximum lateral movement, right maximum lateral movement, total lateral movement (LM), vertical movement (VM), anteroposterior maximum movement, maximum lateral movement (MLM), and total maximum lateral movement (TMLM). RESULTS: The treatment produced increased TMLM, a significant increase of the MLM opposite the crossbite, no statistically significant difference of the crossbite side, and a significant decrease of LM. Left crossbite children showed significant decreases in the LM after treatment; right crossbite children showed significant increases in VM after treatment. CONCLUSIONS: Functional crossbite treatment with selective adjustment and occlusal guiding changed masticatory cycle patterns, producing larger horizontal and vertical planes. The cycles became more symmetric in the frontal plane, increasing the tendency of bilateral alternate mastication. PMID- 17418713 TI - Relationship between cortical bone formation on mandibular condyles and alternation of the magnetic resonance signals characteristic of growth. AB - INTRODUCTION: The purpose of this study was to elucidate the significance of double contour-like structures (DCLS) relative to the maturity of mandibular condyles and the possible constituents of DCLS. We evaluated the relationship between the alternation of DCLS and the ossification on the top of mandibular condyles. METHODS: In 36 subjects, aged 9 to14 years at the first examination, magnetic resonance imaging characteristics of the mandibular condyles were obtained from fast spin echo intermediate-weighted images and T2-weighted images; 2 or 3 years later, magnetic resonance images were again obtained from 23 of the same 36 subjects. RESULTS: A significant relationship was found between the absence of DCLS and the continuity of void-signal lines on the tops of the mandibular condyles. DCLS disappeared, and the void-signal lines on the tops of the mandibular condyles became continuous as the children grew. The top of DCLS was seen as a hypersignal on T2-weighted images. CONCLUSIONS: Alternations of DCLS might be a useful criterion for evaluating the maturity of mandibular condyles in children. Possible constituents of DCLS might be hypervascular loose fibrous tissue and a proliferation of cartilage accompanied by active ossification on the tops of the mandibular condyles. PMID- 17418714 TI - Morphometric evaluation of soft-tissue profile shape. AB - INTRODUCTION: Soft-tissue facial outline has been studied by conventional cephalometric methods, and differences between the 2 sexes have been identified, mainly related to size and timing of growth. However, shape per se was not sufficiently evaluated, especially regarding variability, age-related changes, and sexual dimorphism. The purpose of this study was to evaluate shape variability and sexual dimorphism of the soft-tissue outline by using morphometric methods. METHODS: Pretreatment lateral cephalograms from 170 consecutive patients (82 male, 88 female) aged 7 to 17 years were used. Fifteen skeletal and 22 soft-tissue landmarks were digitized and processed with Procrustes superimposition and principal component analysis. The principal components (PCs) of the soft-tissue shape were analyzed in relation to age and sex. RESULTS: The first 8 PCs explained approximately 90% of the total shape variability. The first coefficient (PC1) related to lip, nose, and chin prominence and included 36% of total shape variability. It was significantly correlated to age, but with a low coefficient of determination (r2 = 13%). The second coefficient (PC2) related to facial convexity and explained 18% of shape variability. The next 2 coefficients were mainly related to lower lip shape. Statistically significant sexual dimorphism was detected, but the overall shape differences between the average profiles of boys and girls were minor and barely detectable visually. Shape dimorphism was present both before and after the age of 12 years. CONCLUSIONS: Shape variability related mainly to relative lip protrusion, convexity of the face, and lower lip shape. Shape differences between the sexes seemed to exist even before the pubertal growth spurt, but they were small. Age changes in shape appeared more significant. PMID- 17418715 TI - Soft-tissue changes during facial growth in skeletal Class II individuals. AB - INTRODUCTION: In orthodontic treatment, a harmonious soft-tissue profile can be difficult to achieve; the thickness of the soft tissues can vary greatly, and changes with growth and treatment can be difficult to predict. The purpose of this study was to investigate changes in the thickness and the length of the soft tissues of the nose, upper and lower lips, and chin due to growth from 6 to 16 years of age. METHODS: Cephalograms of 36 subjects (22 boys, 15 girls) with skeletal Class II malocclusions were analyzed. They participated in the Burlington Growth Study, and longitudinal records had been collected at ages 6, 9, 12, 14, and 16 years. The cephalograms were digitized and analyzed with Dentofacial Planner Plus (version 2.0; Dentofacial Software, Toronto, Ontario, Canada) software. Statistical analysis was performed with the nonparametric Friedman test, and comparisons among means were made with 1-way ANOVA software. RESULTS: All structures showed some growth at all stages. The nose showed the greatest increases in thickness and length in both sexes. There was sexual dimorphism at 16 years, with higher values for boys. Upper lips tended to reduce in the girls. Upper lip length showed slight increases, and base of the upper lip showed small increases for both sexes. The lower lip had moderate increases in all measurements, and its thickness showed sexual dimorphism at almost all ages. The soft tissues of the chin increased in both thickness and length, with no sexual dimorphism. PMID- 17418716 TI - Evaluation of 3 occlusal indexes: Eismann index, Eismann-Farcnik index, and index of orthodontic treatment need. AB - INTRODUCTION: The aim of this study was to compare the validity, reliability, and time needed to use the Eismann index, the Eismann-Farcnik (EF) index, and the index of orthodontic treatment need (IOTN). METHODS: One hundred pretreatment study casts of adolescent patients in the permanent dentition period (mean age, 15.4 years; SD, 0.4 year; 53 girls, 47 boys) were used. Three examiners trained and calibrated in the use of the Eismann and EF indexes and the IOTN scored the casts using the 3 indexes. A panel of 10 orthodontists individually rated the same casts for degree of malocclusion severity. The mean rating of the panel on the severity of malocclusion was used as the gold standard for evaluating the validity of the 3 indexes. Furthermore, the agreement of the 3 indexes in assessing the severity of malocclusion was evaluated. Ten randomly selected casts were reevaluated for the reliability study. Each evaluation was timed and rounded off to the nearest half minute. RESULTS: The intraclass correlation coefficient indicated excellent agreement of the EF index with the ratings of the orthodontists (0.819), good agreement for the Eismann index (0.742), and moderate agreement for the IOTN (0.597). Receiver operating characteristic curves were plotted for the 3 indexes. The diagnostic accuracy, as determined by the area under the curve, was found to be the best for the EF index, followed by the Eismann index and the IOTN. Intraexaminer and interexaminer reliability was high (>0.75) for all 3 indexes. The most time-consuming methods were the EF (27.10 minutes) and the Eismann (26.86 minutes) indexes; the IOTN (1.97 minutes) was the least time-consuming method. CONCLUSIONS: All 3 indexes are valid and reliable methods for assessing malocclusion severity. The EF index is the most time consuming method, but it had the best agreement with clinical orthodontic diagnosis. PMID- 17418717 TI - Heritability of facial proportions and soft-tissue profile characteristics in Turkish Anatolian siblings. AB - INTRODUCTION: The purpose of this study was to determine the possible effects of genetic factors on facial proportions and soft-tissue profile characteristics in Turkish Anatolian siblings. METHODS: The material consisted of lateral and posteroanterior cephalometric head radiographs of 138 siblings (70 women, 68 men) living 15 years or more in the province of Erzurum in eastern Turkey. The subjects were required to have completed their pubertal growth spurts and to have received no previous orthodontic or prosthodontic treatment. The heritability assessments of craniofacial and soft-tissue measurements were undertaken according to narrow-sense heritability. For the calculation and evaluation of heritability estimate values, the mixed-model least-squared and maximum likelihood computer program model type II was used for statistical analysis. RESULTS: According to the narrow-sense heritability estimate values, the measurements of total depth index, soft-tissue chin thickness, soft-tissue facial angle, Merrifield angle, and Holdaway angle showed the highest heritability coefficients (P <.001). Total height index, anterior height index, facial width index, upper to lower facial height index, and lower lip-E line measurements demonstrated moderate heritability values (P<.01). The upper depth index had a low but statistically significant heritability value (P <.05). However, no statistically significant heritability coefficient in upper lip-E line measurement was found. CONCLUSIONS: Turkish Anatolian siblings have similar facial features regarding facial proportions and soft-tissue measurements. PMID- 17418718 TI - Prevalence and distribution of dental anomalies in orthodontic patients. AB - INTRODUCTION: The purpose of this study was to determine the prevalence of developmental dental anomalies in the Turkish population. METHODS: The study was based on the dental casts, intraoral photographs, and panoramic radiographs of 3043 Turkish children (1658 girls, 1385 boys) who had orthodontic treatment at the Department of Orthodontics at the University of Ankara between 1978 and 2003. These patients were examined for 8 developmental dental anomalies: fusion, gemination, microdontia, macrodontia, oligodontia, hypodontia, hyperdontia, and amelogenesis imperfecta. The percentages of these anomalies were assessed in the whole group, and the sexes were compared. RESULTS: It was found that 5.46% of the total group had at least 1 developmental dental anomaly. The distribution by sex was 70 boys (5.05%) and 96 girls (5.79%). CONCLUSIONS: Hypodontia is the most common developmental dental anomaly in the Turkish population, followed by microdontia. PMID- 17418719 TI - The effect of fluoride administration on rat serum osteocalcin expression during orthodontic movement. AB - INTRODUCTION: Osteocalcin is a bone protein that has been used to mark bone turnover. The precise role of this protein in bone remodeling has not been fully elucidated; however, it was shown to be instrumental in the activation of osteoclastic bone resorption via its effect on osteoblasts. Fluoride influences bone growth by acting as a mitogenic agent for osteoblasts. In this study, we used a rodent model to determine the effects of fluoride administration on systemic osteocalcin during orthodontic tooth movement. METHODS: Thirty-two 8 week-old female Wistar rats were divided into 4 groups. The first experimental group (n = 10) was fed a normal laboratory diet and distilled water (MilliQ). The second experimental group (n = 10) had the same diet with the addition of fluoridated water. All animals in both groups had 100-g nickel-titanium coil springs secured to their mandibular incisors and left first molars to instigate orthodontic tooth movement for 14 days. The third (n = 6) and fourth (n = 6) groups were the controls. These animals did not undergo orthodontic tooth movement and were given fluoridated (100 ppm sodium fluoride) and nonfluoridated water ad libitum. Phlebotomy was performed via the lateral tail vein before placement of the orthodontic appliances, and final blood samples were obtained via cardiac puncture after the rats were killed 2 weeks later. Serum was isolated from the samples after centrifuging at each stage of phlebotomy, and an enzyme linked immunosorbent assay (ELISA) was subsequently performed to determine osteocalcin levels in the various sample groups. RESULTS: Orthodontic tooth movement with high, continuous forces was found to have a positive, statistically significant correlation with serum osteocalcin expression in the rodent model. The use of fluoride as a variable was found to increase the mean osteocalcin concentration, but this was not statistically significant. CONCLUSIONS: Further understanding of the biological implications of increased osteocalcin expression requires additional research into the local expression of this protein in the gingival crevice during orthodontic movement. PMID- 17418720 TI - Evaluation of antimicrobial and physical properties of orthodontic composite resin modified by addition of antimicrobial agents--an in-vitro study. AB - INTRODUCTION: This study was designed to investigate the antimicrobial and physical properties of orthodontic composite resin modified by the addition of antimicrobial agents. METHODS: Unite bonding adhesive (3M Unitek, Monrovia, Calif), a chemically cured composite resin, modified by the addition of benzalkonium chloride (BAC), chlorhexidine, and triclosan in concentrations of 0.1 wt%, 0.2 wt%, and 0.3 wt%, respectively, was formed into tablets for testing. The tablets of each group and a control group were subjected to the agar plate diffusion test to measure the zone of bacterial inhibition. In addition, they were incubated for 24 hours in tryptic soy broth inoculated with Streptococcus mutans and examined for bacterial growth. A total of 80 extracted premolars were collected and divided into 2 sets of 40 teeth each. Stainless steel Begg brackets were bonded by using control and experimental composites. A universal testing machine was used to determine shear bond strength. The first set of teeth was tested after 24 hours and the second set after 25 days of storage in distilled water. Time-dependent release of antimicrobial agents from the modified composites was monitored spectrophotometrically. RESULTS AND CONCLUSIONS: The findings indicated that (1) addition of BAC to the orthodontic composite resin enhanced its antimicrobial properties, (2) there was no significant difference in the bond strengths of the control and the experimental resin tested after 24 hours and 25 days, and (3) maximum release of BAC from the modified resin was higher than that of chlorhexidine or triclosan, and was much higher than the minimum inhibitory concentration level. PMID- 17418721 TI - Argon laser vs conventional visible light-cured orthodontic bracket bonding: an in-vivo and in-vitro study. AB - INTRODUCTION: Many advantages of argon lasers have been reported, including high speed orthodontic adhesive curing and less enamel demineralization. The purpose of this study was to compare bond strengths after curing with the argon laser (10 seconds) and the conventional curing light (40 seconds) in vivo and in vitro. METHODS: Four premolars from each of 23 volunteers were randomly assigned to either the argon laser group or the conventional light group for the in-vivo study. Shear bond strengths were measured after 14 days with custom-designed debonding pliers. In-vitro bond strengths were measured by using 4 premolars from each of 25 volunteers. Shear bond strength was measured after 14 days of thermocycling with the same protocol as the in-vivo study. Adhesive remnant index scores (ARI) were determined. RESULTS: No significant differences were found in bond strengths according to curing method, dental arch, or sex. In-vivo results were significantly lower (P < .05) than in-vitro results. A significant (P < .05) difference in ARI scores between the curing methods was determined; no significant correlation between mean bond strengths and ARI scores was determined. CONCLUSIONS: Bond strength for argon laser curing is comparable to conventional light curing and is sufficient for clinical applications. Although the argon laser left more adhesive on the tooth surfaces on debonding, there was no increase in enamel surface fractures. PMID- 17418722 TI - Esthetic segmental retraction of maxillary anterior teeth with a palatal appliance and orthodontic mini-implants. AB - Placing orthodontic mini-implants allows clinicians to use simple and esthetic appliances to retract anterior teeth. This usage is reported here in a patient with lip protrusion and mild crowding. The anterior teeth were splinted on the lingual side and retracted by an elastomeric chain connected to orthodontic mini implants without the use of an archwire or brackets. After space closure, brackets were bonded for detailing individual teeth. The desired movement of the anterior teeth was achieved by changing the application point of the retraction force and adjusting the line of force. PMID- 17418723 TI - Distalization of maxillary molars with the bone-supported pendulum: a clinical study. AB - A modified pendulum appliance with 2 endosseus screws for anchorage in the palatal area was used for maxillary molar distalization in each of 15 patients (average age, 13 +/- 2.1 years). Study models and lateral and panoramic x-rays were taken at the beginning and end of the movement to record the dental and skeletal changes. The mean treatment time was 7.8 +/- 1.7 months, the average distal movement of the maxillary molars was 6 mm, and the inclination was 11.3 degrees +/- 6.2 degrees. The second premolars were distalized an average of 4.85 +/- 1.96 mm with inclinations of 8.6 degrees +/- 5 degrees. The maxillary anterior teeth were retruded 0.5 +/- 1.33 mm and palatally inclined 2.5 degrees +/- 2.98 degrees. The mandibular plane rotated posteriorly 1.27 degrees +/- 1.1 degrees. No loss of dental anchorage was observed during the distal movement. PMID- 17418724 TI - Anterior open bite with temporomandibular disorder treated with titanium screw anchorage: evaluation of morphological and functional improvement. AB - Skeletal anterior open bite is one of the most challenging malocclusions to correct because it is difficult to establish absolute anchorage for molar intrusion with traditional orthodontic mechanics. In addition, patients with anterior open bite sometimes have signs and symptoms of temporomandibular disorders (TMD). In this article, we report the successful treatment of a patient with severe skeletal anterior open bite and TMD; we used titanium screw anchorage. The patient, a woman, age 19 years 11 months, had an open bite of -4.0 mm and increased anterior lower facial height. The titanium screws were implanted in the mandible, and intrusion force was provided with elastic chains for 6 months. After active treatment for 36 months, her mandibular first molars were intruded about 3.0 mm, and good occlusion was achieved. Her retrognathic chin and convex profile were improved both by upward rotation of the mandible and advancement genioplasty with vertical reduction. After treatment, the TMD signs and symptoms were reduced, and improvements of both function and occlusion were achieved. Molar intrusion with titanium screw anchorage might be a useful treatment option to improve function, occlusion, and facial esthetics in patients with severe anterior open bite and TMD. PMID- 17418725 TI - Orthodontics in 3 millennia. Chapter 14: surgical adjuncts to orthodontics. AB - Around 1970, after overcoming obstacles related to anesthesia, infection, and blood supply, orthognathic surgeons come into their own. The history of cleft lip and palate treatment has a much earlier beginning because a deformed infant evokes a strong desire to intervene. Angle's belief that orthodontists can grow bone finally came to fruition with the advent of distraction osteogenesis, which developed from the limb-lengthening procedures of Ilizaroff in Russia. Now distraction osteogenesis has replaced osteotomies in many applications. PMID- 17418726 TI - An orphan subspecialty, or whither pediatric cardiac anesthesia? PMID- 17418727 TI - Extravasation of albumin after cardiopulmonary bypass in newborns. AB - OBJECTIVE: The systemic inflammatory response to cardiopulmonary bypass (CPB) possibly increases microvascular permeability to plasma proteins, leading to capillary leak syndrome. The study was conducted to elucidate any protein leakage in newborns using Evans blue dye as tracer. DESIGN: Prospective controlled study. SETTING: University-affiliated heart center. PARTICIPANTS: Eleven neonates with transposition of the great arteries. INTERVENTIONS: Plasma interleukin-6 (IL-6), IL-10, fractional escape rate (FER) of an intravenous bolus of Evans blue, and colloid osmotic pressure (COP) were assessed before and after surgery (statistics: median and 25th-75th percentile, Friedman's 2-way analysis of variance, and Wilcoxon matched-pairs signed-rank test [before and after surgery]). MEASUREMENTS AND MAIN RESULTS: All patients had an uneventful intraoperative course. The demographic and operative data were age 11 (10-13) days, body weight 3.2 (3.0-3.3) kg, CPB time 132 (123-144) minutes, and aortic cross-clamp time 66 (64-78) minutes. The proinflammatory IL-6 increased 60-fold and the anti-inflammatory IL-10 only 3-fold after CPB. FER, however, was not changed, whereas COP was significantly reduced after CPB. CONCLUSIONS: In contrast to the expectation, the escape rate of Evans blue, reflecting the extravasation of albumin, was not increased after CPB. However, reduced COP, hypothermia, and also a reduced lymphatic drainage may contribute to edema formation. The present data do not support the hypothesis of a capillary leak after CPB in newborns. PMID- 17418728 TI - Effects of hemodilution on outcome after modified Blalock-Taussig shunt operation in children with cyanotic congenital heart disease. AB - OBJECTIVE: Cyanotic congenital heart diseases (CCHD) with secondary polycythemia and hyperviscosity state are associated with a reduction in blood flow, stagnation of blood, and thrombosis. Sufficient hemodilution in cyanotic children results in higher blood flow and significant reductions in perioperative blood loss. The aim of this study was to investigate similar beneficial effects of hemodilution in preventing shunt thrombosis and decreasing postoperative blood loss after modified Blalock-Taussig (BT) shunt operations in children with CCHD. DESIGN: Prospective, randomized, controlled study. SETTING: Cardiac center of a tertiary care, referral hospital. PARTICIPANTS: Fifty children with CCHD undergoing modified BT shunt operations. INTERVENTIONS: Patients were randomized into 2 groups. The study group (n = 25) received a calculated amount of 6% hydroxyethyl starch (200/0.5) solution to bring down the hematocrit to 45%, whereas the control group (n = 25) received 5% dextrose solution intraoperatively as per the authors' normal protocol. MEASUREMENTS AND MAIN RESULTS: Effects of hemodilution on shunt patency, postoperative blood loss at 24 hours, blood and blood component usage, and re-exploration rates were recorded. The shunt patency rate was significantly higher in the study group than the control group (100% and 84%, respectively, p < 0.05). Postoperative blood loss at 24 hours was significantly higher in the control group than in the study group (14.4 +/- 11.8 mL/kg and 9.9 +/- 8 mL/kg, respectively, p < 0.05). The number of recipients and the amount of blood and blood components administered were higher in the control group, but they were not statistically significant. The re-exploration rate (for excessive postoperative chest-tube drainage) was significantly higher in the control group than the study group (12% and 0%, respectively, p < 0.05). CONCLUSION: Hemodilution in CCHD patients undergoing modified BT shunt surgery has beneficial effects including improved shunt patency because of higher blood flow through the graft and less postoperative blood loss, which may be attributed to the lower viscosity produced by hemodilution. PMID- 17418729 TI - Perioperative serum interleukins in neonates with hypoplastic left-heart syndrome and transposition of the great arteries. AB - OBJECTIVE: The primary study objective was to examine the impact of diagnosis on the inflammatory response in neonates with congenital heart disease undergoing cardiac surgery. The secondary objective was to study the impact of the inflammatory response on postoperative outcome in these neonates. DESIGN: Observational study. SETTING: Tertiary care children's hospital heart center. PATIENTS: Neonates with hypoplastic left-heart syndrome (HLHS) undergoing stage I repair and patients with transposition of the great arteries (TGA) undergoing arterial switch operation. MEASUREMENTS AND MAIN RESULTS: There were 24 neonates with HLHS and 21 neonates with TGA. Serum samples to measure interleukin (IL)-6 and -10 were obtained before and after CPB at 1, 3, 6, and 24 hours postoperatively. Time to extubation, intensive care unit (ICU) length of stay, and peritoneal fluid drainage were compared between the groups. Serum IL-6 and IL 10 concentrations increased after CPB when compared to the preoperative concentration. Preoperative concentrations of IL-6 were significantly elevated in the HLHS group (HLHS: 32 [21.1, 69.6] pg/mL v TGA: 7.2 [3.6, 22.5] pg/mL [median, 25th, and 75th percentile], p = 0.003) and remained elevated immediately after CPB, and at 3 and 6 hours postoperatively. The IL-10 to IL-6 ratio was lower in the HLHS group preoperatively and immediately after CPB. ICU length of stay was significantly longer in the HLHS group (TGA 4 [3-6] days v HLHS 6 [5-8] days, p = 0.031). Mortality in the HLHS group (4/24) was associated with significantly higher IL-6 postoperatively (IL-6 immediately postoperatively: HLHS survivors 59.9 [34.3, 65.7] pg/mL v nonsurvivors 98.7 [94.4, 104.5] pg/mL, p < 0.011). CONCLUSIONS: All neonates with TGA or HLHS have a significant inflammatory response after CPB. Neonates with HLHS have evidence of an activated inflammatory response before CPB, which remains significant in the postoperative period. Accelerated interleukin expression and an abnormal cytokine balance correlate with longer time to extubation, longer ICU length of stay, and increased peritoneal fluid volume. PMID- 17418730 TI - The release of systemic inflammatory mediators is independent of cardiopulmonary bypass temperature. AB - OBJECTIVE: The aim of this study was to investigate the effect of systemic CPB temperature on the production of the key mediators of the systemic inflammatory response to coronary artery bypass graft (CABG) surgery. DESIGN: Randomized clinical study. SETTING: University hospital. PARTICIPANTS: Thirty patients undergoing first-time CABG surgery. INTERVENTIONS: The patients were randomized to hypothermic (32 degrees C, n = 15) or normothermic (36 degrees C, n = 15) CPB. MEASUREMENTS AND MAIN RESULTS: Plasma interleukin (IL)-6, IL-8, IL-10, C-reactive protein (CRP), cortisol, and neutrophils were measured the day before operation, at closure of the sternum, and 4, 16, and 44 hours later. The cytokine, CRP, cortisol, and neutrophil responses were independent of temperature during CPB with peak concentrations of IL-10 at closure of the sternum followed by IL-6, IL 8, cortisol, neutrophils, and finally CRP. A correlation between maximal plasma concentrations of IL-10 and cortisol was seen in both groups after surgery (p = 0.02). Drainage after surgery was lower after normothermic CPB (p=0.02), with no difference in the requirement for blood transfusion. All patients were discharged from the intensive care unit within 24 hours after surgery. CONCLUSIONS: The release of systemic inflammatory mediators after cardiac surgery was independent of mild hypothermia (32 degrees C) versus normothermia (36 degrees C) during CPB. PMID- 17418731 TI - The air-leak test is not a good predictor of postextubation adverse events in children undergoing cardiac surgery. AB - OBJECTIVE: The air-leak test is recommended as a method of assessing the appropriate size of an uncuffed endotracheal tube (ETT) in children. The authors' primary objective was to determine whether the air-leak test would predict adverse events and reintubations after the removal of the ETT in children who have undergone cardiac surgery. DESIGN: Prospective, observational, clinical study. SETTING: University tertiary care hospital. PATIENTS: Ninety-four children <10 years of age undergoing elective cardiac surgery requiring cardiopulmonary bypass surgery. INTERVENTIONS: The attending anesthesiologist assessed air-leak pressure after intubation in the operating room (OR). In addition, the air-leak test was performed in 42 patients before extubation in the pediatric intensive care unit (PICU). The incidence of adverse events and the number of failed extubations were recorded after removal of the ETT. MEASUREMENTS AND MAIN RESULTS: Eleven of the 94 patients were excluded from the study. Four (4.3%) of the patients died in the PICU before extubation, and 7 patients were excluded for other reasons. The median age of the 83 children was 0.9 years (range 0.01-9.6 years). The total incidences of postextubation adverse events and failed extubations were 30.1% and 8.4%, respectively. An audible air leak < or =25 cmH(2)O airway pressure during the OR phase or before removal of the ETT during the PICU recovery phase had no significant predictive value for the incidence of adverse events (p = 0.63) or reintubations (p = 1.0). The patients undergoing simple and complete operations compared with more complex and incomplete operations had significantly fewer postextubation adverse events (p = 0.03). Neonates did not have a higher risk for postextubation adverse events (p = 0.64) or reintubations (p = 0.26) than older children. CONCLUSION: The air-leak test did not predict an increased risk for postextubation adverse events and reintubations in children undergoing elective congenital heart surgery. PMID- 17418732 TI - Effects of escalating doses of sildenafil on hemodynamics and gas exchange in children with pulmonary hypertension and congenital cardiac defects. AB - OBJECTIVE: Sildenafil (Viagra, Pfizer) is being increasingly used to treat pulmonary hypertension in children. However, there are limited data available to suggest dosage regimens. The purpose of this study was to determine the effects of escalating doses of sildenafil on hemodynamics and gas exchange in children with pulmonary hypertension because of congenital cardiac defects. DESIGN: Prospective, observational study. SETTING: Pediatric intensive care unit in a tertiary care children's hospital. PARTICIPANTS: Ten children with pulmonary hypertension because of congenital cardiac defects who were in the intensive care unit and on nitric oxide after cardiac surgery. INTERVENTION: Patients received sildenafil every 4 hours via a gastric tube in incremental doses of 0.5 mg/kg, 1 mg/kg, 1.5 mg/kg, and 2.0 mg/kg along with nitric oxide during their stay in the intensive care unit until they were extubated. Hemodynamic and arterial blood gas measurements were taken before (baseline) and 60 minutes after the administration of sildenafil. MEASUREMENTS AND MAIN RESULTS: All doses of sildenafil caused significant reduction in pulmonary artery pressure with no significant effect on systemic arterial and central venous pressures. Arterial partial pressure of oxygen was decreased after a 2.0 mg/kg dose of sildenafil but not significantly. No significant differences were found among the 4 doses. CONCLUSION: For the treatment of pulmonary hypertension in children with congenital cardiac defects, a 0.5 mg/kg dose of sildenafil every 4 hours is therapeutically as effective as a 2.0 mg/kg dose every 4 hours. However, a large dose-ranging and pharmacokinetic study of sildenafil in children with pulmonary hypertension because of congenital cardiac defects is needed to validate the safety and efficacy of the dose-range and dosing interval suggested by this study. PMID- 17418733 TI - Cardiopulmonary bypass for lung transplantation in cystic fibrosis: pilot evaluation of perioperative outcome. AB - OBJECTIVE: The purpose of this study was to determine whether cardiopulmonary bypass (CPB) reduces the incidence of perioperative graft infection after lung transplantation in adults with cystic fibrosis (CF). DESIGN: Retrospective and observational. SETTING: University hospital. PARTICIPANTS: Adults with CF who underwent lung transplantation (1998-2003). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Cohort size was 26: group A (n = 10) who underwent CPB for implantation of both lungs, group B (n = 8) who underwent CPB only for implantation of the second lung, and group C (n = 8) who did not undergo CPB. The 3 cohort subgroups were similar (p > 0.05) in demographics, preoperative lung function, and anesthetic management. Group A had a lower incidence of perioperative pneumonia (p = 0.02). CPB exposure increased transfusion (B > A > C) of fresh frozen plasma and platelets but not packed red blood cells. There were no differences (p > 0.05) in clinical outcome as reflected by duration of mechanical ventilation, tracheal re-intubation, re-exploration for bleeding, sepsis, primary graft dysfunction, renal dysfunction, length of stay, and mortality. CONCLUSIONS: CPB is associated with decreased incidence of early graft infection after lung transplantation for adult CF when used for implantation of both lungs. This may be because of improved decontamination of the operative field before graft implantation. PMID- 17418734 TI - Integrity of the cerebral blood-flow response to hyperoxia after cardiopulmonary bypass. AB - OBJECTIVE: In this study, the hypothesis that cardiopulmonary bypass (CPB) alters the cerebral blood flow (CBF) vasoconstrictive response to hyperoxia was tested. DESIGN: A prospective, observational study was conducted. SETTING: The study was conducted at a single university hospital. PARTICIPANTS: Subjects were patients who presented for cardiac surgery with CPB. INTERVENTIONS: CBF was measured before and after CPB in 12 subjects while breathing 21% O(2) and 100% O(2). CBF was measured by using continuous arterial spin labeling (CASL) perfusion magnetic resonance imaging. Arterial pO(2) (mmHg), pCO(2) (mmHg), hemoglobin (Hgb), and oxygen content (CaO(2)) were also measured. MEASUREMENTS AND MAIN RESULTS: Mean age of the 12 subjects was 63 +/- 16 years. Hgb decreased from 12.0 (+/-2.4) g/dL to 9.2 (+/-2.9) g/dL postoperatively (p = 0.008). CBF increased by 39%, from 37.2 (+/-10.8) mL/100 g/min to 49.2 (+/-14.3)mL/100 g/min postoperatively (p = 0.01). In response to the hyperoxic challenge CBF decreased by 8.0 (+/-7.1) mL/100 g/min (21%) preoperatively and by 9.4 (+/-6.4) mL/100 g/min (19%) postoperatively (p = 0.58). By using multiple regression, the contribution of CPB to the hyperoxic CBF response (DeltaCBF) was evaluated, while controlling for other potentially important covariates known to influence CBF, including age, baseline CBF on 21% O(2), and changes in arterial pO(2), pCO(2), and CaO(2). CPB state was not found to be a significant covariate in controlling the CBF response to hyperoxia. CONCLUSIONS: CPB does not impair the CBF response to hyperoxia. PMID- 17418735 TI - Blood concentrations of enflurane before, during, and after hypothermic cardiopulmonary bypass. AB - OBJECTIVE: The purpose of this study was to determine blood concentrations of enflurane delivered via a membrane oxygenator during hypothermic cardiopulmonary bypass (CPB) with changes in the input enflurane concentration and temperature and to characterize the pharmacokinetics of enflurane washout during and after CPB. DESIGN: Blood enflurane concentrations were measured by gas chromatography before, during, and after CPB by using mean delivered enflurane concentrations of 0.5% v/v (group 1, n = 5), 0.8% (group 2, n = 7), and 1% (group 3, n = 14). SETTING: The investigation was performed in a teaching hospital setting. PARTICIPANTS: Twenty-six patients undergoing cardiac surgery requiring hypothermic CPB. INTERVENTIONS: Variations in input enflurane concentration in different patients plus blood sampling from the arterial side of the circuit for enflurane assay. MEASUREMENTS AND MAIN RESULTS: Median (25th and 75th percentiles) pre-CPB blood enflurane concentrations were 48 (25-50) mg/L, 52 (47 56) mg/L, and 115 (90-143) mg/L in groups 1 (0.5% v/v), 2 (0.8% v/v), and 3 (1% v/v), respectively. During hypothermia (28 degrees C) corresponding enflurane concentrations were 44 (31-53) mg/L, 56 (45-62) mg/L, and 145 (109-203) mg/L, respectively. For groups 1 and 2, there were no significant changes in blood enflurane compared with the corresponding pre-CPB value. However, for group 3, cooling resulted in a significant increase (p = 0.006) in blood enflurane. In all groups, enflurane concentrations after rewarming were similar to those in the pre CPB period. CONCLUSIONS: It is concluded that exposure to enflurane concentrations greater than 0.8% during CPB can result in high blood concentrations. PMID- 17418736 TI - Inhibition of thrombin during reperfusion improves immediate postischemic myocardial function and modulates apoptosis in a porcine model of cardiopulmonary bypass. AB - OBJECTIVE: Transient left-ventricular dysfunction because of myocardial reperfusion injury is a significant problem after cardiac surgery, but the underlying complex pathophysiology is still poorly understood. The authors studied early functional recovery of the postischemic myocardium and explored potential effects of thrombin inhibition on procoagulatory, proinflammatory, and proapoptotic features of myocardial ischemia-reperfusion injury. DESIGN: A randomized, blinded study. SETTING: University research laboratory. SUBJECTS: Porcine model. INTERVENTIONS: Twenty pigs undergoing 60 minutes of aortic clamping and 75 minutes of normothermic cardiopulmonary bypass (CPB) received an intravenous bolus of r-hirudin (10 mg, 0.4mg/kg, n = 10) or placebo (n = 10) 15 minutes before aortic declamping, followed by a 135-minute intravenous infusion of r-hirudin (3.75 mg, 0.15 mg/kg/h) or placebo. MEASUREMENTS AND MAIN RESULTS: Hemodynamic parameters were measured before CPB, after weaning from CPB, and at 30, 60, 90, and 120 minutes after aortic declamping. Blood was sampled, and myocardial biopsies were taken before CPB, just before aortic declamping, during reperfusion, and after 120 minutes of reperfusion to measure thrombin antithrombin complexes and to quantitate leukocyte infiltration (myeloperoxidase activity) for histologic evaluation and detection of apoptosis with caspase-3 and the TUNEL method. The r-hirudin group showed significantly higher stroke volume and cardiac output than the control group at 60 minutes and at 90 minutes after aortic declamping (p < 0.05). Microthrombosis was not observed in either group, indicating sufficient anticoagulation and excluding intravascular clots as explanations for LV dysfunction in the current experiment. Instead, ample myocardial activation of inflammation was present, but only a trend of r-hirudin associated anti-inflammatory effect was observed. Compared with the controls, TUNEL-positive myocytes were detected significantly less frequently in the r hirudin group (0.05 +/- 0.06 v 0.13 +/- 0.07 TUNEL-positive nuclei %, p = 0.042). CONCLUSIONS: The improved cardiac recovery in the r-hirudin group during reperfusion after cardioplegia-induced cardiac arrest was associated with significant differences in cardiomyocyte apoptosis and anti-inflammatory effects. Thus, in clinical cardiac surgery, inhibition of reperfusion- induced thrombin may offer beneficial effects by mechanisms other than direct anticoagulation. PMID- 17418737 TI - Universal leukocyte reduction of transfused red cells does not provide benefit to patients undergoing cardiac surgery. AB - OBJECTIVE: A policy of universal leukocyte reduction of the blood supply in Finland was implemented at the beginning of 2003. The aim of the present evaluation was to determine the potential role of leukocyte-reduced red blood cells in decreasing postoperative infections. DESIGN: A retrospective cohort study. SETTING: A major university clinic. PARTICIPANTS: Consecutive patients undergoing cardiac surgery during the years 2002 and 2003. INTERVENTIONS: Transfused patients received either buffy-coat-depleted red blood cells before leukocyte reduction (n = 782) or leukocyte-reduced red blood cells after leukocyte reduction (n = 632). MEASUREMENTS AND MAIN RESULTS: The evaluated outcome parameters were culture-proven postoperative infections, 90-day mortality, and length of stay in the intensive care unit. The percentage of patients transfused with red blood cells (56% v 53%, p = 0.16) and amounts of transfused red blood cells (4.3 +/- 6.7 [3.0] units v 4.3 +/- 6.6 [2.0] units, means +/- standard deviation [median], p = 0.48) were comparable between the study groups (buffy-coat-depleted group and leukocyte-reduced group, respectively). The 90-day mortality (6.6% v 6.3%, p = 0.28), the length of intensive care stay (3.6 +/- 4.7 [2.0] days v 4.3 +/- 7.1 [2.0] days, p = 0.34), and the number of patients with culture-proven infections (8.8% v 10.9%, p = 0.19) were unchanged after universal leukocyte reduction. In multivariate comparisons, the leukocyte reduction was not associated with culture-proven postoperative infections and 90-day mortality. CONCLUSION: No beneficial effect of the universal leukocyte reduction in cardiac surgery was found for culture positive infection rates, 90-day mortality, or length of intensive care stay. PMID- 17418738 TI - A systematic approach to substernal epicardial echocardiographic examination. AB - OBJECTIVE: The importance of echocardiography in the perioperative management of cardiac surgical patients is widely appreciated. A modified mediastinal drain has been developed, which allows the introduction of a standard TEE probe in a closed ended sleeve coupled with the drain to permit epicardial echocardiographic imaging after chest closure (substernal epicardial echocardiography [SEE]). The aim of the present study was to develop a standardized and comprehensive SEE examination sequence to allow repeatable examinations with a single movement of the TEE probe inside the drain. DESIGN: Prospective observational protocol. SETTING: Tertiary care university hospital. PARTICIPANTS: Ten adult patients undergoing elective cardiac surgery. INTERVENTIONS: Twenty-three SEE examinations in 10 patients undergoing elective myocardial revascularization to develop a standard examination sequence. MEASUREMENTS AND MAIN RESULTS: The examination sequence includes 11 views with all the structures relevant for postoperative monitoring. The entire sequence is performed with a single in-out movement of the transesophageal probe to minimize discomfort to patients and the risk of damaging the tube. CONCLUSIONS: This new approach to the perioperative monitoring of cardiac surgical patients represents an option for patients in whom TEE is contraindicated or multiple examinations are anticipated because SEE examinations can be performed without the need for sedation in awake patients. PMID- 17418739 TI - An anaphylactic reaction after primary exposure to an aprotinin test dose in a child with a severe milk allergy. PMID- 17418740 TI - Deep hypothermic circulatory arrest in a patient with osteogenesis imperfecta. PMID- 17418741 TI - Horseshoe lung secondary to hypoplastic left lung for pneumonectomy. PMID- 17418742 TI - Anesthetic management of combined heart-liver transplantation. PMID- 17418743 TI - Mediastinal mass diagnosed with intraoperative transesophageal echocardiography. PMID- 17418744 TI - The management of a patient with tracheal compression undergoing combined resection of an anterior mediastinal mass and aortic valve replacement with coronary artery bypass graft surgery: utility of the laryngeal mask airway and Aintree intubation catheter. PMID- 17418745 TI - Management of a patient with right ventricular drainage cannula obstruction after biventricular assist device implantation. PMID- 17418746 TI - Intraoperative hyperthermic intrathoracic chemotherapy for pleural extension of pseudomyxoma peritonei. PMID- 17418747 TI - Use of continuous venovenous hemofiltration for reversal of anticoagulation with lepirudin post-cardiopulmonary bypass in a patient with heparin-induced thrombocytopenia after heart transplantation. PMID- 17418748 TI - Hyponatremia and cardiopulmonary bypass. PMID- 17418749 TI - Iatrogenic aortic incompetence after mitral valve replacement. PMID- 17418750 TI - Mechanical ventilation affects pulmonary inflammation in cardiac surgery patients: the role of the open-lung concept. PMID- 17418751 TI - Pioneering cardiac anesthesia: the life and work of Emerson Moffitt, MD. PMID- 17418752 TI - Case 2--2007: Systemic air embolization after termination of cardiopulmonary bypass. PMID- 17418753 TI - Pro: Aprotinin should be used in coronary artery bypass graft surgery with cardiopulmonary bypass. PMID- 17418754 TI - Con: Aprotinin should not be used in cardiac surgery with cardiopulmonary bypass. PMID- 17418755 TI - Left ventricular endocardial irregularity: evaluation using epicardial echocardiography. PMID- 17418756 TI - A widened mediastinum in a patient with a left parasternal stab wound. PMID- 17418757 TI - The possible mechanism of apnea after institution of cardiopulmonary bypass in conscious patients. PMID- 17418758 TI - Acute back pain and paresthesia after femoral venous catheter placement. PMID- 17418759 TI - Transpharyngeal imaging of the carotid artery and internal jugular vein: possible roles in cerebral perfusion management during adult aortic arch repair. PMID- 17418760 TI - Another technique of internal jugular vein catheterization. PMID- 17418761 TI - The use of rigid bronchoscopy for bronchial stenting in patients with tracheal stenosis. PMID- 17418763 TI - [Surgical management of the thumb with intrinsic palsy]. AB - The long term disability resulting from paralysis of the muscle that control the thumb depends largely of the management (surgical and physiotherapic) of these problems that must be considered in the context of functional disability, and so it is possible to consider specific tendon transfers to improve opposition. This work dealt with paralysis of the thenar muscles. It is helpful to treat such a problem to consider restoring of the anteposition, rotation and adduction of the first metacarpal. In paralysis of the abductor pollicis brevis (APB) and opponents pollicis (OP) it is possible to use as a motor transfer the tendon of extensor pollicis brevis (EPB) or extensor indicis proprius (EPB) or occasionally by palmaris longus (PL). When the flexor pollicis brevis is paralysed, transfer of the flexor sublimis ot the third or the ring finger is used as described by Bunnel. When all thenar muscles are paralysed is very important to assess the range of movement of the trapeziometacarpal joint as well as the presence of any contracture of the first web space. The stability of the metacarpophalangeal and interphalangeal joints must be assessed with pinch grip between thumb and index finger. Careful preoperative assessment of remaining function allows accurate diagnosis and thus what tendon transfer is possible and what other procedure are necessary like arthrodesis of the MP joint, opening of the first web, or stabilization of the IP. The management of these problems is difficult and many factors must be taken into consideration like age, aetiology and experience of the surgeon. PMID- 17418764 TI - The anatomy and vascularity of the lunate: considerations applied to Kienbock's disease. AB - PURPOSE: The purpose of this study was to assess the anatomy and vascularity of the lunate. The genesis of lunatomalacia requires some combination of vascular risk and mechanical predisposition. The findings will be correlated with the major existing theories of the cause of Kienbock's disease. METHODS: We studied 27 cadaver upper limbs using latex injection and the Spalteholz technique. We investigated the blood supply to the lunate. In 24 wrists we evaluated the incidence and distribution of anatomic features, arthrosis, and soft tissue lesions. We correlated the lunate morphology and ligaments disruptions with the arthritic changes. RESULTS: The lunate morphology results as classified by Antuna Zapico were five type I (20.8%), 18 type II (75%) and one type III (4.2%). The lunate was found to have a separate facet for the hamate in 11 cases (45.8%). The most common size of the facet was found to be 3 mm (range, 3-6 mm). Arthrosis was identified with most frequency in the radius (88.2%) and lunate (94.1%). The triangular fibrocartilage complex was found torn in 58.3%, the lunotriquetral interosseous ligament was torn in 20.8% and the scapholunate interosseous ligament (SLIL) was torn in 54.2% of the wrists. There was a correlation between the presence of arthrosis at the hamate and the presence of a lunate facet (P=0.027) and a correlation between the presence of a tear in the SLIL and arthrosis in the scaphoid (P=0.002). The nutrient vessels entered the lunate through the dorsal and volar poles in all the specimens. The dorsal intercarpal and radiocarpal arches supply blood to the lunate from a plexus of vessels located directly over the lunate's dorsal pole. Vessels entered the dorsal aspect of the lunate through one to three foramina. One to five nutrient vessels were observed entering the volar pole through various ligament insertions, including the ligament of Testut-Kuentz (radio-scapho-lunate (RSL) ligament) and the radiolunate triquetrum ligament (or dorsoradial carpal ligament) and ulnar lunate triquetral ligament. CONCLUSIONS: The lunate had consistent dorsal and palmar arteries entering the bone in all the specimens. The blood supply and foramina number is greater in the volar pole of the lunate than the dorsal pole. The lunate blood supply comes from different ligaments. In the etiopathogeny of Kienbock's disease it is possible that an acute or chronic, traumatic or non traumatic injury of the vessel bearing ligaments, particularly because of their structure and the location of the RSL ligament, may have an important role in the appearance of lunate necrosis. PMID- 17418765 TI - [Dynamic external traction system for management of distal displaced fractures of the head of the proximal phalanx]. AB - INTRODUCTION: Phalangeal fractures are generally underestimated injuries, but result in pain, stiffness, instability, degenerative arthritis and loss of finger function. Treatment varies from open reduction with internal fixation giving perfect anatomical reduction but a risk of side effects, to orthopaedic treatment with variable results. Unstable fractures must be fixed rigidly to minimize joint immobilization. It is this concept of solid fixation and early mobilization, which directed us in the choice of dynamic external distraction in the management of these fractures. MATERIAL AND METHOD: The authors present a prospective study of twelve patients treated for unstable fractures of the proximal interphalangeal joint by closed reduction and dynamic external distractor, we describe this simple device created with pins and rubber bands and examine the functional clinical results. RESULTS: The average follow-up period was 7.5 months. We only noted one pin-track infection. Three patients complained of pain at postoperative follow up. In this series, the total active motion varied from 70 degrees to 120 degrees (average 94.16 degrees), the mean lack of extension was 5.6 degrees (from 0 to 15 degrees). Ten patients had good mobility and were satisfied. DISCUSSION: Different traction devices for the management of displaced intraarticular phalangeal fractures have been reported. Most of them are complex to construct and expensive. The traction system is simple to assemble, economical and it allows early active motion of the affected digits. We consider that it is an effective mode of treatment for the management of complex intraarticular phalangeal fractures. PMID- 17418766 TI - [Addition-subtraction osteotomy combined with ligamentoplasty for symptomatic trapezial dysplasia with instability]. AB - INTRODUCTION: Trapezometacarpal instability with trapezial dysplasia is an incapacitating condition long before radiological changes appear. Most of the patients are young and demanding. Treatment is often conservative. Surgical treatment options are relatively classic, starting with a ligamentoplasty of the base of the first metacarpal or a tenotomy of the transarticular accessory slip of the abductor pollicis longus. Other options such as arthrodesis and arthroplasty have not been so successful in this situation. METHOD: The idea arose to combine two techniques described previously. In 1973 Wilson published his technique of abduction osteotomy of the first metacarpal as a treatment for basal osteoarthritis of the thumb. In 2002, Kapandji and Heim published their opening wedge osteotomy of the trapezium in order to correct the slope of a dysplastic trapezium. The combination of both of these techniques avoids closing of the first web by utilising the abduction osteotomy of the first metacarpal. Moreover, the opening wedge osteotomy corrects the dysplastic lateral rim of the trapezium by correcting its slope. RESULTS: We have been performing this technique since 2001 and 2005 in six patients. The preliminary results are encouraging, 5 of them were able to resume their work. A fair result was achieved in a young patient. DISCUSSION: The described technique is relatively easy and has the advantage of preserving the trapezometacarpal joint. In addition, it leaves the door open for other techniques if it eventually should fail in the long term. PMID- 17418767 TI - [The Foucher's "kite-flap" (12 cases)]. AB - The "kite-flap" is a cutaneous island flap containing a triple pedicle. It is harvested from the dorsum of the index and is intended to cover skin-loss on the thumb. The purpose of our study was to evaluate 12 "kite-flaps" performed over the course of 11 years. The skin defects were on the dorsum of the distal phalanx of the thumb in 3 cases, on the dorsum of both the distal and proximal phalanges in 4 cases and on the volar aspect of the thumb in 5 cases. The associated lesions consisted of combined bone and skin in one case treated by bone graft with external fixation, and one case of septic arthritis of the interphalangeal joint treated by arthrodesis. The results of our series are very good after an average of 2 years follow-up. We have however had 4 cases of cold intolerance, 2 cases of superficial sepsis and one case of shrinkage of the first commissure. It appears that the "kite-flap" is the best solution for cover of simple or complex skin loss of the thumb. Its technical performance is easy, and it gives durable, sensate and stable skin cover. PMID- 17418768 TI - [Primary flexor tendon ruptures of the little finger within the carpal tunnel. Proposed classification based on six clinical cases and review of the literature]. AB - Six cases of primary flexor tendon rupture of the little finger within the carpal tunnel are presented. All of them were attritional ruptures caused by a rough hook of the hamate. These ruptures are rare. They have their place among the flexor tendon ruptures of the wrist and the palm. Ultrasound may by helpful to determine the level of rupture. The surgical treatment consists of excision of the hook of the hamate and tendon repair. PMID- 17418769 TI - [Testing of pronation and supination strength in the outpatient setting: a preliminary study]. AB - Dynamometric studies of grip strength are widely used in clinics. We present a clinic dynamometer that can be used to measure pronation and supination moments while the subject being tested is holding a cylindrical handle in neutral pronosupination. One hundred volunteers without any previous history of upper extremity injury were tested in pronation and supination in order to provide a database of strength values and to study variations with respect to sex, age, dominance, weight and height. PMID- 17418770 TI - [Intramedullary stabilisation of hematometacarpal fracture-dislocation. A case report]. AB - We report a case of a dorsal coronal fracture of the hamate associated with an unstable hematometacarpal dislocation in a 15 year old patient. An accurate assessment was done only after a 3D-CT was performed. Open reduction and intramedullary fixation yielded a very good result. PMID- 17418771 TI - [Pseudoaneurysm of the deep palmar arch due to infective endocarditis]. AB - The authors report a case of mycotic pseudoaneurysm of the deep palmar arch in a complicated case of Staphylococcus aureus infective endocarditis. They review the literature to discuss the physiopathology, diagnosis and treatment of this rare complication. PMID- 17418772 TI - [Bifocal fracture of sternal end of clavicle-pseudodislocation of sternoclavicular joint. Report of a case]. AB - We report an unusual case of bifocal fracture of the sternal part of clavicle, combining a metaphysodiaphysal fracture of the sternal end of the clavicle, a proximal physeal fracture, and a ligament avulsion of sternoclavicular joint, without neurovascular damage. Its surgical repair used an osteosynthesis with K wires and tension band wiring and synthetic ligamentoplasty of the anterior and posterior sternoclavicular ligaments, without any reconstruction of the costoclavicular ligament. The clinical and radiographic results at six months are presented and the operative technique used in this case of pseudodislocation of sternoclavicular joint is discussed and compared to those previously published. PMID- 17418773 TI - [An unusual manifestation of osteo-articular, tendinous and nervous involvement secondary to tophaceous gout of the hand. A report of two cases]. AB - Tophaceous gout of the hand is a rare condition, but possible in chronic cases. We report two cases (three hands) with a long history of hyperuricemia and serious lesions due to deposition of sodium urate crystals. In these cases we point out several fundamental aspects: the abnormal size of the tophi, the destruction of triquetum and pisiform, the digital joint instability due to ligamentous lesions, the tenosynovitis and digital flexion contracture secondary to intratendinous deposition of urates, and the presence of a double ipsilateral tunnel syndrome of the median and ulnar nerves at the wrist. After neurolysis and tophus resection, a rapid and very substantial relief of paraesthesiae was seen. Recovery of partial active wrist and finger movements required a variety of surgical interventions. The results are analysed after 2 years for the first case, and 8 months for the second. PMID- 17418774 TI - [Double dislocation of the fifth metacarpal]. AB - A case of simultaneous dorsal dislocation of the metacarpophalangeal and carpometacarpal joints in the little finger is presented. The patient required an open reduction of the metacarpophalangeal joint, the carpometacarpal joint was reduced simultaneously. Thirty month post-operatively there was no subluxation of either joint. PMID- 17418775 TI - [Bipolar dislocation of the forearm]. AB - The authors report the case of a 23 year old patient, who presented the following combination of injuries after a fall, namely a divergent dislocation of the elbow, a fracture of the radial head, a fracture of the ulna and a perilunate dislocation with scaphoid fracture. The entire injury thus represented a bipolar dislocation of the forearm. The emergency management consisted of a closed reduction of the elbow together with osteosynthesis of the ulna, internal fixation of the scaphoid with a compression screw, and scaphocapitate K wiring. After 26 months, the function of the elbow was satisfactory, and the range of motion of the wrist was from 50 degrees of flexion to 40 degrees of extension. Satisfactory healing of the scaphoid was observed. This combination of injuries has only rarely been reported in the literature. PMID- 17418776 TI - Scientists on the move. AB - In today's global world, scientists are increasingly mobile and the traditional view of "brain drain" is being replaced by "brain circulation." Countries such as China and India are tapping into a global pool of science talent attracting not only their own citizens back from abroad but also scientists from the US and Europe. PMID- 17418777 TI - How a home-grown T cell receptor interacts with a foreign landscape. AB - The ability of a T cell receptor (TCR) to directly recognize foreign (allogeneic) major histocompatibility complex (MHC) molecules underlies T cell-mediated rejection in patients receiving allogeneic organ transplants. In this issue, Colf et al. (2007) reveal that instead of mimicking the interactions formed with a self MHC, a single TCR adopts a completely different strategy to recognize a foreign MHC. PMID- 17418778 TI - A blend of two circadian clocks, seasoned to perfection. AB - The daily activity of the fruit fly Drosophila is controlled by both a "morning" and an "evening" circadian clock. In this issue Stoleru et al. (2007) demonstrate that day length determines which clock dominates the neural circuitry governing circadian behavior. Thus, these findings suggest a mechanism by which the system for circadian timing adapts to changes in the seasons to impose appropriate rhythms of daily activity. PMID- 17418779 TI - An atomic switch for memory. AB - Molecular genetic techniques are beginning to bring about a detailed understanding of the biochemical processes underlying complex cognitive phenomena such as memory, with investigations approaching atomic-level resolution. An excellent example of this is provided in this issue of Cell by Costa-Mattioli et al. (2007), whose results implicate a single protein dephosphorylation event in the control of long-term memory formation in mice. PMID- 17418780 TI - IKKalpha at the crossroads of inflammation and metastasis. AB - The development of solid tumors is regulated by dynamic interactions between evolving neoplastic cells and their microenvironment. Luo et al. (2007) recently demonstrated that tumor-infiltrating immune cells expressing RANKL induce activation and nuclear localization of IKKalpha in prostatic epithelial tumor cells. This leads to repression of maspin, a critical suppressor of metastasis, and thus commits malignant prostatic epithelial cells to a metastatic fate. PMID- 17418781 TI - A method to the madness of N-glycan complexity? AB - Cell-surface glycoprotein receptors have varying numbers of N-glycan sites. In this issue of Cell, Lau et al. (2007) report that increasing intracellular UDP GlcNAc leads to increased branching of N-glycans, increased receptor association with cell-surface galectin-3, and enhanced signaling. They also show that the kinetics of this response differ between growth-promoting receptors, which have 8 16 N-glycans, and those that induce growth arrest, which have very few N-glycans, suggesting that hexosamine flux may regulate the transition from growth to arrest. PMID- 17418782 TI - Waiting to inhale: HIF-1 modulates aerobic respiration. AB - The hypoxia-inducible factor HIF-1 is known to promote anaerobic respiration during low oxygen conditions (hypoxia). In this issue, Fukuda et al. (2007) expand the range of HIF-1's functions by showing that it modulates aerobic respiration as well. PMID- 17418783 TI - Development in motion: helper T cells at work. AB - In mammals, helper T cells orchestrate defense against diverse pathogens. However, these warriors of the immune system can also result in self-inflicted injury culminating in autoimmune and allergic diseases. Recent findings--such as the discovery of the Th17 lineage--have revealed additional complexity in the fates chosen by helper T cells and have begun to reshape our view of how signaling and transcriptional networks generate appropriate and inappropriate immunity. PMID- 17418784 TI - Mighty Piwis defend the germline against genome intruders. AB - Piwis are a germline-specific subclass of the Argonaute family of RNA interference (RNAi) effector proteins that are associated with a recently discovered group of small RNAs (piRNAs). Recent studies in Drosophila and zebrafish directly implicate Piwi proteins in piRNA biogenesis to maintain transposon silencing in the germline genome (Brennecke et al., 2007; Gunawardane et al., 2007; Houwing et al., 2007). This function may be conserved in mice as loss of Miwi2, a mouse Piwi homolog, leads to germline stem cell and meiotic defects correlated with increased transposon activity (Carmell et al., 2007). PMID- 17418785 TI - Bcl-2 and Bcl-XL regulate proinflammatory caspase-1 activation by interaction with NALP1. AB - Caspases are intracellular proteases that cleave substrates involved in apoptosis or inflammation. In C. elegans, a paradigm for caspase regulation exists in which caspase CED-3 is activated by nucleotide-binding protein CED-4, which is suppressed by Bcl-2-family protein CED-9. We have identified a mammalian analog of this caspase-regulatory system in the NLR-family protein NALP1, a nucleotide dependent activator of cytokine-processing protease caspase-1, which responds to bacterial ligand muramyl-dipeptide (MDP). Antiapoptotic proteins Bcl-2 and Bcl X(L) bind and suppress NALP1, reducing caspase-1 activation and interleukin-1beta (IL-1beta) production. When exposed to MDP, Bcl-2-deficient macrophages exhibit more caspase-1 processing and IL-1beta production, whereas Bcl-2-overexpressing macrophages demonstrate less caspase-1 processing and IL-1beta production. The findings reveal an interaction of host defense and apoptosis machinery. PMID- 17418786 TI - Communication between distant sites in RNA polymerase II through ubiquitylation factors and the polymerase CTD. AB - Transcriptional arrest triggers ubiquitylation of RNA polymerase II (RNAPII). We mapped the yeast RNAPII ubiquitylation sites and found that they play an important role in elongation and the DNA-damage response. One site lies in a protein domain that is unordered in free RNAPII, but ordered in the elongating form, helping explain the preferential ubiquitylation of this form. The other site is >125 Angstroms away, yet mutation of either site affects ubiquitylation of the other, in vitro and in vivo. The basis for this remarkable coupling was uncovered: an Rsp5 (E3) dimer assembled on the RNAPII C-terminal domain (CTD). The ubiquitylation sites bind Ubc5 (E2), which in turn binds Rsp5 to allow modification. Evidence for folding of the CTD compatible with this mechanism of communication between distant sites is provided. These data reveal the specificity and mechanism of RNAPII ubiquitylation and demonstrate that E2s can play a crucial role in substrate recognition. PMID- 17418787 TI - A role for Piwi and piRNAs in germ cell maintenance and transposon silencing in Zebrafish. AB - Piwi proteins specify an animal-specific subclass of the Argonaute family that, in vertebrates, is specifically expressed in germ cells. We demonstrate that zebrafish Piwi (Ziwi) is expressed in both the male and the female gonad and is a component of a germline-specifying structure called nuage. Loss of Ziwi function results in a progressive loss of germ cells due to apoptosis during larval development. In animals that have reduced Ziwi function, germ cells are maintained but display abnormal levels of apoptosis in adults. In mammals, Piwi proteins associate with approximately 29-nucleotide-long, testis-specific RNA molecules called piRNAs. Here we show that zebrafish piRNAs are present in both ovary and testis. Many of these are derived from transposons, implicating a role for piRNAs in the silencing of repetitive elements in vertebrates. Furthermore, we show that piRNAs are Dicer independent and that their 3' end likely carries a 2'O-Methyl modification. PMID- 17418788 TI - Natively unfolded nucleoporins gate protein diffusion across the nuclear pore complex. AB - Nuclear pore complexes (NPCs) form aqueous conduits in the nuclear envelope and gate the diffusion of large proteins between the cytoplasm and nucleoplasm. NPC proteins (nucleoporins) that contain phenylalanine-glycine motifs in filamentous, natively unfolded domains (FG domains) line the diffusion conduit of the NPC, but their role in the size-selective barrier is unclear. We show that deletion of individual FG domains in yeast relaxes the NPC permeability barrier. At the molecular level, the FG domains of five nucleoporins anchored at the NPC center form a cohesive meshwork of filaments through hydrophobic interactions, which involve phenylalanines in FG motifs and are dispersed by aliphatic alcohols. In contrast, the FG domains of four peripherally anchored nucleoporins are generally noncohesive. The results support a two-gate model of NPC architecture featuring a central diffusion gate formed by a meshwork of cohesive FG nucleoporin filaments and a peripheral gate formed by repulsive FG nucleoporin filaments. PMID- 17418789 TI - Protein translocation is mediated by oligomers of the SecY complex with one SecY copy forming the channel. AB - Many proteins are translocated across the bacterial plasma membrane by the interplay of the cytoplasmic ATPase SecA with a protein-conducting channel, formed from the evolutionarily conserved heterotrimeric SecY complex. Here, we have used purified E. coli components to address the mechanism of translocation. Disulfide bridge crosslinking demonstrates that SecA transfers both the signal sequence and the mature region of a secreted substrate into a single SecY molecule. However, protein translocation involves oligomers of the SecY complex, because a SecY molecule defective in translocation can be rescued by linking it covalently with a wild-type SecY copy. SecA interacts through one of its domains with a nontranslocating SecY copy and moves the polypeptide chain through a neighboring SecY copy. Oligomeric channels with only one active pore likely mediate protein translocation in all organisms. PMID- 17418790 TI - HIF-1 regulates cytochrome oxidase subunits to optimize efficiency of respiration in hypoxic cells. AB - O(2) is the ultimate electron acceptor for mitochondrial respiration, a process catalyzed by cytochrome c oxidase (COX). In yeast, COX subunit composition is regulated by COX5a and COX5b gene transcription in response to high and low O(2), respectively. Here we demonstrate that in mammalian cells, expression of the COX4 1 and COX4-2 isoforms is O(2) regulated. Under conditions of reduced O(2) availability, hypoxia-inducible factor 1 (HIF-1) reciprocally regulates COX4 subunit expression by activating transcription of the genes encoding COX4-2 and LON, a mitochondrial protease that is required for COX4-1 degradation. The effects of manipulating COX4 subunit expression on COX activity, ATP production, O(2) consumption, and reactive oxygen species generation indicate that the COX4 subunit switch is a homeostatic response that optimizes the efficiency of respiration at different O(2) concentrations. Thus, mammalian cells respond to hypoxia by altering COX subunit composition, as previously observed in yeast, but by a completely different molecular mechanism. PMID- 17418791 TI - Complex N-glycan number and degree of branching cooperate to regulate cell proliferation and differentiation. AB - The number of N-glycans (n) is a distinct feature of each glycoprotein sequence and cooperates with the physical properties of the Golgi N-glycan-branching pathway to regulate surface glycoprotein levels. The Golgi pathway is ultrasensitive to hexosamine flux for the production of tri- and tetra-antennary N-glycans, which bind to galectins and form a molecular lattice that opposes glycoprotein endocytosis. Glycoproteins with few N-glycans (e.g., TbetaR, CTLA-4, and GLUT4) exhibit enhanced cell-surface expression with switch-like responses to increasing hexosamine concentration, whereas glycoproteins with high numbers of N glycans (e.g., EGFR, IGFR, FGFR, and PDGFR) exhibit hyperbolic responses. Computational and experimental data reveal that these features allow nutrient flux stimulated by growth-promoting high-n receptors to drive arrest/differentiation programs by increasing surface levels of low-n glycoproteins. We have identified a mechanism for metabolic regulation of cellular transition between growth and arrest in mammals arising from apparent coevolution of N-glycan number and branching. PMID- 17418792 TI - How a single T cell receptor recognizes both self and foreign MHC. AB - alphabeta T cell receptors (TCRs) can crossreact with both self- and foreign- major histocompatibility complex (MHC) proteins in an enigmatic phenomenon termed alloreactivity. Here we present the 2.35 A structure of the 2C TCR complexed with its foreign ligand H-2L(d)-QL9. Surprisingly, we find that this TCR utilizes a different strategy to engage the foreign pMHC in comparison to the manner in which it recognizes a self ligand H-2K(b)-dEV8. 2C engages both shared and polymorphic residues on L(d) and K(b), as well as the unrelated QL9 and dEV8 peptide antigens, in unique pair-wise contacts, resulting in greater structural complementarity with the L(d)-QL9 complex. In the structure of an engineered, high-affinity 2C TCR variant bound to H-2L(d)-QL9, the "wild-type" TCR-MHC binding orientation persists despite modified TCR-CDR3alpha interactions with peptide. Thus, a single TCR recognizes two globally similar, but distinct ligands by divergent mechanisms, indicating that receptor-ligand crossreactivity can occur in the absence of molecular mimicry. PMID- 17418793 TI - The mammalian Golgi regulates numb signaling in asymmetric cell division by releasing ACBD3 during mitosis. AB - Mammalian neural progenitor cells divide asymmetrically to self-renew and produce a neuron by segregating cytosolic Numb proteins primarily to one daughter cell. Numb signaling specifies progenitor over neuronal fates but, paradoxically, also promotes neuronal differentiation. Here we report that ACBD3 is a Numb partner in cell-fate specification. ACBD3 and Numb proteins interact through an essential Numb domain, and the respective loss- and gain-of-function mutant mice share phenotypic similarities. Interestingly, ACBD3 associates with the Golgi apparatus in neurons and interphase progenitor cells but becomes cytosolic after Golgi fragmentation during mitosis, when Numb activity is needed to distinguish the two daughter cells. Accordingly, cytosolic ACBD3 can act synergistically with Numb to specify cell fates, and its continuing presence during the progenitor cell cycle inhibits neuron production. We propose that Golgi fragmentation and reconstitution during cell cycle differentially regulate Numb signaling through changes in ACBD3 subcellular distribution and represent a mechanism for coupling cell-fate specification and cell-cycle progression. PMID- 17418794 TI - Distinct target-derived signals organize formation, maturation, and maintenance of motor nerve terminals. AB - Target-derived factors organize synaptogenesis by promoting differentiation of nerve terminals at synaptic sites. Several candidate organizing molecules have been identified based on their bioactivities in vitro, but little is known about their roles in vivo. Here, we show that three sets of organizers act sequentially to pattern motor nerve terminals: FGFs, beta2 laminins, and collagen alpha(IV) chains. FGFs of the 7/10/22 subfamily and broadly distributed collagen IV chains (alpha1/2) promote clustering of synaptic vesicles as nerve terminals form. beta2 laminins concentrated at synaptic sites are dispensable for embryonic development of nerve terminals but are required for their postnatal maturation. Synapse specific collagen IV chains (alpha3-6) accumulate only after synapses are mature and are required for synaptic maintenance. Thus, multiple target-derived signals permit discrete control of the formation, maturation, and maintenance of presynaptic specializations. PMID- 17418795 TI - eIF2alpha phosphorylation bidirectionally regulates the switch from short- to long-term synaptic plasticity and memory. AB - The late phase of long-term potentiation (LTP) and memory (LTM) requires new gene expression, but the molecular mechanisms that underlie these processes are not fully understood. Phosphorylation of eIF2alpha inhibits general translation but selectively stimulates translation of ATF4, a repressor of CREB-mediated late-LTP (L-LTP) and LTM. We used a pharmacogenetic bidirectional approach to examine the role of eIF2alpha phosphorylation in synaptic plasticity and behavioral learning. We show that in eIF2alpha(+/S51A) mice, in which eIF2alpha phosphorylation is reduced, the threshold for eliciting L-LTP in hippocampal slices is lowered, and memory is enhanced. In contrast, only early-LTP is evoked by repeated tetanic stimulation and LTM is impaired, when eIF2alpha phosphorylation is increased by injecting into the hippocampus a small molecule, Sal003, which prevents the dephosphorylation of eIF2alpha. These findings highlight the importance of a single phosphorylation site in eIF2alpha as a key regulator of L-LTP and LTM formation. PMID- 17418796 TI - The Drosophila circadian network is a seasonal timer. AB - Previous work in Drosophila has defined two populations of circadian brain neurons, morning cells (M-cells) and evening cells (E-cells), both of which keep circadian time and regulate morning and evening activity, respectively. It has long been speculated that a multiple oscillator circadian network in animals underlies the behavioral and physiological pattern variability caused by seasonal fluctuations of photoperiod. We have manipulated separately the circadian photoentrainment pathway within E- and M-cells and show that E-cells process light information and function as master clocks in the presence of light. M-cells in contrast need darkness to cycle autonomously and dominate the network. The results indicate that the network switches control between these two centers as a function of photoperiod. Together with the different entraining properties of the two clock centers, the results suggest that the functional organization of the network underlies the behavioral adjustment to variations in daylength and season. PMID- 17418797 TI - SnapShot: mammalian TRP channels. PMID- 17418798 TI - Quantitative profiling of epoxyeicosatrienoic, hydroxyeicosatetraenoic, and dihydroxyeicosatetraenoic acids in human intrauterine tissues using liquid chromatography/electrospray ionization tandem mass spectrometry. AB - A reversed-phase liquid chromatography negative ion electrospray tandem mass spectrometry (LC/ESI-MS/MS) method was developed and validated to quantify a range of physiologically relevant eicosanoids, including 5,6-, 8,9-, 11,12-, and 14,15-epoxyeicosatrienoic acids (EETs); 5-, 8-, 9-, 12-, and 15 hydroxyeicosatetraenoic acids (HETEs), and 5,6-, 8,15-, and 12,20 dihydroxyeicosatetraenoic acids (DiHETEs) in human intrauterine tissues. A solid phase extraction method was employed to extract the eicosanoids, and gradient LC separation was performed on a Kromasil C(18) column. Mass spectrometric detection was performed by multiple reaction monitoring over a 31-min run time. The calibration curves were linear over the range of 4-400pmol/g tissue, and the intra- and interday precision and accuracy were within a coefficient of variation of 2.0 to 27.4% and 4.6 to 17.9%, respectively. The lower limit of quantitation was 1.0pmol/g tissue. The method was applied successfully to the characterization and quantitation of eicosanoids in the different compartments of human intrauterine tissues. Our results demonstrate significantly greater amounts of HETEs than of either the EETs or DiHETEs (P<0.001), irrespective of tissue type. Specifically, the metabolite 12-HETE was significantly more abundant (P<0.001) than all other HETEs. Of the EET metabolites, 5,6-EET predominated (P<0.001). A significant negative correlation between EETs and HETEs for all tissues (rho= 0.390, P<0.001) was identified, implying a biological feedback mechanism between these two arachidonate metabolite classes. PMID- 17418799 TI - Investigation of a protein expression profile by high-resolution bidimensional electrophoresis of Trypanosoma cruzi epimastigotes. PMID- 17418800 TI - Identification of cis-regulatory elements and trans-acting proteins of the rat carbohydrate response element binding protein gene. AB - Carbohydrate response element binding protein (ChREBP) is a transcription factor that activates liver glycolytic and lipogenetic enzyme genes in response to high carbohydrate diet. Here we report the transcriptional regulatory mechanisms for the rat ChREBP gene. Firstly, we determined the transcription initiation site and the nucleotide sequences of the rat ChREBP promoter region encompassing approximately 900bp from the ATG initiation codon. Reporter gene assays demonstrated that the major positive regulatory region exists in the nucleotide sequence between -163 and -32 of the ChREBP gene. This region contains a cluster of putative transcription factor binding elements that consist of two specificity protein 1 (Sp1) binding sites (-66 to -50 and -93 to -78), a sterol regulatory element (-101 to -110), and two nuclear factor-Y (NF-Y) binding sites (-23 to -19 and -131 to -127). Mutations introduced into these sites caused marked reduction of ChREBP promoter activities. Functional synergisms were observed between Sp1/NF Y and Sp1/sterol regulatory element-binding protein. Additionally, electrophoretic mobility shift assays and chromatin immunoprecipitation assays demonstrated that these factors bound to these elements. Thus, we conclude that functional synergisms between these transcription factors are critical for ChREBP gene transcription. PMID- 17418801 TI - Mass isotopomer study of anaplerosis from propionate in the perfused rat heart. AB - Anaplerosis from propionate was investigated in rat hearts perfused with 0-2mM [(13)C(3)]propionate and physiological concentrations of glucose, lactate, and pyruvate. The data show that when the concentration of [(13)C(3)]propionate was raised from 0 to 2mM, total anaplerosis increased from 5% to 16% of the turnover of citric acid cycle intermediates. Then, [(13)C(3)]propionate abolished anaplerosis from endogenous substrates, glucose, lactate, and pyruvate. Also, while the contents of propionyl-CoA and methylmalonyl-CoA increased with [(13)C(3)]propionate concentration, the content of succinyl-CoA decreased, presumably via activation of succinyl-CoA hydrolysis by a decrease in free CoA. Under our conditions, [(13)C(3)]propionate was a purely anaplerotic substrate since there was no labeling of mitochondrial acetyl-CoA, reflected by the labeling of the acetyl moiety of citrate. PMID- 17418802 TI - Investigation of the ligand spectrum of human sterol carrier protein 2 using a direct mass spectrometry assay. AB - Sterol carrier protein 2 (SCP2) has been investigated by nearly native electrospray ionisation mass spectrometry in the presence of long chain fatty acyl CoAs (LCFA-CoAs) and carnitine derivatives of equivalent fatty acid chain length (LCFA-carnitines). Four SCP2 constructs were compared to examine the influence of the N-terminal presequence and the C-terminal peroxisomal targeting signal on ligand binding. Removal of N- or C-terminal residues did not influence ligand binding. The observation that LCFA-CoAs are high affinity ligands for SCP2 was confirmed, while LCFA-carnitines were demonstrated for the first time not to interact with SCP2. LCFA-CoAs formed non-covalent complexes with SCP2 of 2:1 and 1:1 stoichiometry, which could be dissociated by elevating the energy of the ions upon entrance to the mass spectrometer. A fluorescence-competition assay using Nile Red butyric acid confirmed the mass spectrometric observations in solution. The physiological significance of the lack of LCFA-carnitine binding by SCP2 is discussed. PMID- 17418803 TI - Catalytic characterization and cytokine mediated regulation of cytochrome P450 4Fs in rat hepatocytes. AB - Cytochrome P450 (CYP) 4F mediated leukotriene B(4) (LTB(4)) metabolism modulates inflammation during injury and infection. Here we show that in addition to LTB(4), the recombinant rat CYP4Fs catalyze omega-hydroxylations of lipoxin A(4), and hydroxyeicosatetraeonic acids. CYP4F gene regulation studies in primary hepatocytes reveal that pro-inflammatory cytokines interleukin (IL) -1beta, IL-6 and tumor necrosis factor (TNF) -alpha produce a general inductive response whereas IL-10, an anti-inflammatory cytokine, suppresses CYP4F expression. The molecular mechanism behind IL-6 related induction of CYP4F4 and 4F5 is partially signal transducer and activator of transcription 3 (STAT3) dependent. When hepatocytes are subjected to high concentrations of LTB(4) or prostaglandin E(2), lipid mediators of inflammation, only an increase in CYP4F5 mRNA expression is observed. Collectively, the results from isozyme activity and substrate driven CYP4F induction do not support the notion that an autoregulatory pathway could control the excessive concentrations of LTB(4) during an inflammatory challenge to hepatocytes. PMID- 17418804 TI - Does anxiety sensitivity predict symptoms of panic, depression, and social anxiety? AB - This study examined whether the lower-order factors of the Anxiety Sensitivity Index (ASI) exhibited specificity in predicting symptoms of panic, depression, and social anxiety prospectively. This question was addressed using a sample of undergraduates stratified to represent low, medium, and high levels of anxiety sensitivity (AS). It was hypothesized that the physical concerns, mental concerns, and social concerns subscales of the ASI would predict increases in panic, depression, and social anxiety symptoms, respectively, one year later. Results found that the physical concerns subscale predicted increases in both panic and depressive symptoms. Neither the mental concerns nor the social concerns subscales predicted significant variance in any of the Time 2 symptoms. Theoretical implications of these data for AS are discussed. PMID- 17418805 TI - Identification and characterization of the novel human prostate cancer-specific PC-1 gene promoter. AB - Human prostate and colon gene-1 (PC-1, also known as PrLZ) is an androgen regulated, prostate tissue and prostate cancer cells specifically expressed novel gene. The increased expression of PC-1 gene appears to promote prostate cancer cells androgen-dependent (AD) and androgen-independent (AI) growth. To clone and investigate the expression and regulation elements of PC-1 gene may provide insight into the function of PC-1 and develop a new promoter that targets therapeutic genes to the AD and AI prostate cancer cells. The goal of the present study is cloning and characterization of the PC-1 promoter. A series of luciferase constructs that contain various fragments of the PC-1 5'-genomic region were transfected into human prostate cancer cells for promoter transactivation analysis. 5' deletion analysis identified the -1579 bp promoter region was required for the maximal proximal promoter activity; two transcriptional suppression and a positive regulatory region were identified; 4939 bp promoter fragment of the PC-1 gene retained the characteristic of prostate cancer-specific expression and exhibited higher transcription activity than PSA-6 kb promoter in the medium supplemented with steroid-depleted FBS. An androgen response element (ARE) was located in between -345 and -359 bp of the PC 1 5'-untranslated region relative to the translation initiation site. Thus, our studies not only provide molecular basis of PC-1 transcription regulation, but also define a new regulatory sequence that may be used to restrict expression of therapeutic genes to prostate cancer in the prostate cancer gene therapy. PMID- 17418806 TI - Injectable gel with synthetic collagen-binding peptide for enhanced osteogenesis in vitro and in vivo. AB - A synthetic peptide denoted as collagen-binding motif (CBM) was identified from osteopontin (OPN), a multisubunit extracellular matrix (ECM) protein, by enzymatic digestion with chymotrypsin. The aim of this study was to examine the feasibility of identified CBM peptide as an active component of gel type scaffold material in osteogenesis. The binding of CBM peptide to collagen was specific and presented high affinity. Cell adhesion and growth on CBM peptide-immobilized gel were significantly increased as compared with those on gel with control peptide or without peptide. The CBM peptide-immobilized gel increased osteoblastic differentiation, followed by marked bone formation in the rabbit calvarial defect sites at 4 weeks. Taken together, the injectable gel with synthetic CBM peptide has a potential to induce osteogenesis in vitro and in vivo, suggesting its clinical application in bone regeneration procedure. PMID- 17418807 TI - Effect of PPAR-delta agonist on the expression of visfatin, adiponectin, and resistin in rat adipose tissue and 3T3-L1 adipocytes. AB - It has been recently reported that activation of PPAR-delta, by specific agonists or genetic manipulation, alleviates dyslipidemia, hyperglycemia, and insulin resistance in animal models of obesity and type 2 diabetes. The purpose of the present study was to determine whether the PPAR-delta agonist has a direct effect on adipokines in visceral adipose tissue of rats and in cultured adipocytes. We examined the expression of visfatin, adiponectin, and resistin mRNA in visceral adipose tissue of Wistar rats fed a high-fat diet and 3T3-L1 adipocytes treated with PPAR-delta agonist (L-165041). Body weight and biochemical measurements were performed. Rats fed a high-fat diet showed a greater increase in body weight than those fed a standard diet (P<0.05), and treatment with L-165041 (10 mg/kg/day) significantly decreased weight gain (P<0.05). The concentration of total cholesterol was lower, and HDL cholesterol was higher in L-165041-treated rats (P<0.05). In the visceral adipose tissue of L-165041-treated rats, visfatin and adiponectin mRNA levels significantly increased compared to those of the untreated rats (P<0.05). However, the expression of resistin decreased in the L 165041-treated rats. Furthermore, in cultured 3T3-L1 adipocytes, the level of visfatin and adiponectin mRNA was up-regulated in response to L-165041 treatment for nine days. By contrast, resistin mRNA levels were down-regulated by L-165041 treatment. The present study provides a novel evidence to suggest that the PPAR delta agonist has regulatory effects on a variety of adipokines, and these effects might explain some of their metabolic function. PMID- 17418808 TI - LMO4 mRNA stability is regulated by extracellular ATP in F11 cells. AB - LIM only domain protein 4 (LMO4) interacts with many signaling and transcription factors to regulate cellular proliferation, differentiation and plasticity. In Drosophila, mutations in the 3' untranslated region (UTR) of the homologue dLMO cause a gain of function by increasing mRNA stability. LMO4 3'UTR contains several AU-rich elements (ARE) and is highly conserved among vertebrates, suggesting that RNA destabilizing mechanisms are evolutionarily conserved. Here, we found that extracellular ATP stabilized LMO4 mRNA in F11 cells. The LMO4 3'UTR added to a luciferase reporter markedly reduced reporter activity under basal conditions, but increased activity with ATP treatment. Two ARE motifs were characterized in the LMO4 3'UTR. ATP increased binding of HuD protein to ARE1. ARE1 conferred ATP and HuD-dependent mRNA stabilization. In contrast, sequences flanking ARE2 bound CUGBP1 and ATP destabilized this complex. Thus, our results suggest that ATP modulates recruitment of RNA-binding proteins to the 3'UTR to stabilize LMO4 mRNA. PMID- 17418809 TI - Inhibition of breast cancer cell adhesion and bone metastasis by the extracellular adherence protein of Staphylococcus aureus. AB - Bone metastasis is a common sequelae of breast cancer and the interaction of alpha v beta3-integrin with osteopontin (OPN) found in the extracellular matrix of mineralized tissues is implicated in this process. The integrin-dependent proadhesive and promigratory functions of OPN are particularly attributed to the 40 kD N-terminal fragment that derives upon matrix metalloproteinase (MMP) cleavage. Based on the broad repertoire of interactions between Staphylococcus aureus extracellular adherence protein (Eap) and host components, we here characterized Eap to specifically interact with recombinant full-length OPN and the 40 kD N-terminal MMP cleavage fragment, but not with the 32 kD or the 25 kD C terminal fragments of OPN. Eap thereby prevented the OPN/alpha v beta3-integrin interaction, as well as the alpha v beta3-integrin-dependent adhesion of MDA-MB 231 breast cancer cells to full-length OPN or to the 40 kD fragment and the migration of these cells towards OPN. Furthermore, Eap treatment markedly impaired the development of osseous metastasis of human MDA-MB-231 cells in vivo. Taken together, Eap may represent an attractive novel treatment for the prevention of breast cancer bone metastasis. PMID- 17418810 TI - Minimal translation of the tmRNA tag-coding region is required for ribosome release. AB - The trans-translation system in bacteria promotes recycling of stalled ribosomes and targets incomplete peptides for proteolysis. In Escherichia coli, loss of trans-translation function has little effect on growth under normal laboratory conditions. Among the subtle phenotypes of tmRNA-deficient mutants is the inability to plate certain lambda imm(P22) phages. This phenotype is dependent on the ribosome recycling functions of the trans-translation system but is independent of its proteolysis-targeting activity. The experiments described here show that translation of the first (resume) codon of the tmRNA open reading frame by a tRNA is both necessary and sufficient for ribosome recycling. While a variety of sense codons can replace the naturally-occurring GCA alanine codon as the resume codon, both AAA and AAG lysine codons are non-functional resume codons. These results suggest that the main function of tmRNA in releasing stalled ribosomes is to supply a stop codon and so facilitate termination and subsequent ribosome recycling. PMID- 17418811 TI - High level expression and purification of recombinant PEX protein in cultured skeletal muscle cell expression system. AB - Large quantities of recombinant proteins are needed for specific therapeutic and diagnostic applications. To achieve high-level expression in eukaryotic cells, the choice of cell line as well as the expression vector is critical. In this report, we demonstrate that a combination of the skeletal muscle cell line, QM7 and a cytomegalovirus promoter-based expression vector can achieve high-level expression of secretory recombinant proteins in eukaryotic cells. We also screened a serum-free medium containing 3 microg/ml insulin suitable for QM7 differentiation and identified a very potent signal peptide from MMP9, which effectively directs secretion of heterologous proteins. The C-terminal hemopexin like domain of MMP-2, PEX, a powerful candidate for the treatment of diseases associated with neovascularization was expressed in QM7 cells with bioactivity. This skeletal muscle cell-based system may be employed for the production of human proteins of special interests, such as those for structural determination or therapeutical development. PMID- 17418812 TI - Quantitative mRNA analysis of serotonin 5-HT4 receptor isoforms, calcium handling proteins and ion channels in human atrial fibrillation. AB - Serotonin 5-HT(4) receptors are present in human atrial myocytes and have been proposed to contribute to the generation of atrial fibrillation (AF). Here, we quantified 5-HT(4) receptors as well as other key genes involved in cardiac rhythm and contraction in right atrial appendages of patients with chronic AF (CAF) and acute AF (AAF). Right atrial appendages were obtained from eleven patients in sinus rhythm (SR), five with AAF and six with CAF (>12 months). TaqMan real time quantitative RT-PCR was performed on total RNA. Results were normalised to the average of three housekeeping genes, cyclophilin, GADPH and RL 19. The rank order of expression of h5-HT(4) receptors variants was (b)>(a)>(g)>(c) in the group of patients in SR. In AAF, we found a strong decrease in h5-HT(4(b)), h5-HT(4(c),) and h5-HT(4(g)) transcripts. In CAF patients, the mRNA expression level of the h5-HT(4(b)) isoform significantly increased two fold versus SR. A similar increase was reported for beta(1) adrenergic receptor, connexin 43 and the L-type Ca(2+) channel CaCNA1C subunit. Interestingly, CAF was associated with a strong increase in the expression of Na(+)/Ca(2+) exchanger and the voltage-dependent Na(+) channel SCN5A subunit. Our results indicate that h5-HT(4(b)) is the dominant cardiac isoform of human 5 HT(4) receptors and its expression is increased in CAF. These data support the involvement of 5-HT(4) receptors in atrial arrhythmia. PMID- 17418813 TI - Reversible AChE inhibitors in C. elegans vs. rats, mice. AB - We are investigating whether Caenorhabditis elegans could be used as a screen for vertebrates by comparing the responses of components of its cholinergic system to well-characterized toxicants. We assessed whether C. elegans displays similar toxicity as rats and mice to reversible acetylcholinesterase (AChE) inhibitors, and sought to corroborate that the toxicity mechanism is the same. To determine relative potencies, movement-concentration curves were generated, 50th percentiles for movement were located, ranked and compared statistically to rat and mouse oral acute LD50s. The ranking was significantly correlated to rat and mouse rankings (alpha=0.05). We measured a concentration-dependent decrease in AChE activity correlating to a decrease in movement for each carbamate, suggesting that the mechanism of toxicity is the same. Finally, as seen in mammals, inhibition of AChE activity occurred before a movement decrease. The response of C. elegans to carbamate exposure shows significant correlation to rat and mouse data. PMID- 17418814 TI - Endothelin-2/vasoactive intestinal contractor via ROCK regulates transglutaminase 1 on differentiation of mouse keratinocytes. AB - We previously found that endothelin-2/vasoactive intestinal contractor (ET-2/VIC) greatly increased in mouse epidermis after birth. In the present study, we evaluated whether ET-2/VIC expression was associated with the calcium-induced differentiation of cultured mouse keratinocytes. The differentiation induction was revealed by morphological change, cornified envelope (CE) formation, and involucrin and transglutaminase 1 (TG 1) expressions. ET-2/VIC gene expression and peptide production subsequently increased in the induction of the differentiation. We also found that Y-27632, a Rho-associated coiled-coil forming protein serine/threonine kinase (ROCK) inhibitor, suppressed up-regulation of ET 2/VIC gene expression, the induction of morphological change, the CE formation, and TG 1 expression, but not involucrin expression. These results indicate new three findings, (1) ET-2/VIC expression increases and has potential as a differentiation marker, (2) ET-2/VIC expression is mediated by ROCK, and (3) the ROCK regulated TG 1 expression, on the calcium-induced differentiation of mouse keratinocytes. PMID- 17418815 TI - Glutamate induces apoptosis in cultured spiral ganglion explants. AB - Traumatic sound exposure, aminoglycoside antibiotics, cochlea ischemia or traumatic stress leads to an excessive release of glutamate from inner hair cells into the synaptic cleft. The high glutamate concentration can cause a swelling and destruction of the dendrites of spiral ganglion neurons of type I as well as a reduction in the number of neurons. This may be a cause of hearing loss. The mechanism causing the reduction of neurons is still not known. Apoptosis, also called programmed cell death, could be involved. In this study, cultured spiral ganglion explants were incubated with glutamate in high concentrations. Neurite outgrowth was determined and additionally a new method was established for studying the morphology of single spiral ganglion neurons. For the first time it was shown that glutamate induces apoptosis of spiral ganglion neurons, which could be blocked selectively by a caspase-3 inhibitor. This could offer a new therapeutic strategy for hearing disorders. PMID- 17418816 TI - Human spot 14 protein interacts physically and functionally with the thyroid receptor. AB - Spot 14 (S14) is a small acidic protein with no sequence similarity to other mammalian gene products. Its biochemical function is elusive. Recent studies have shown that, in some cancers, human S14 (hS14) localizes to the nucleus and is amplified, suggesting that it plays a role in the regulation of lipogenic enzymes during tumorigenesis. In this study, we purified untagged hS14 protein and then demonstrated, using various biochemical methods, including analytic ultracentrifugation, that hS14 might form a homodimer. We also found several lines of evidence to suggest physical and functional interactions between hS14 and the thyroid hormone receptor (TR). The ubiquitous expression of hS14 in various cell lines and its cell-type-dependent functions demonstrated in this study suggest that it acts as a positive or negative cofactor of the TR to regulate malic enzyme gene expression. These findings provide a molecular rationale for the role of hS14 in TR-dependent transcriptional activation of the expression of specific genes. PMID- 17418817 TI - Promotion of DNA strand breaks, interstrand cross-links and apoptotic cell death in A2780 human ovarian cancer cells by transplatinum planar amine complexes. AB - Cisplatin is one of the primary drugs utilized in the treatment of ovarian cancer. However, despite the initial effectiveness of chemotherapy in suppressing this disease, drug resistance almost invariably develops and cures are relatively rare. While it is generally thought that only compounds of the cis geometry express antitumor activity, a number of transplatinum derivates have shown preclinical promise. The current work investigates the influence of transplanaramine (TPA) compounds of structure trans-[Pt (O(2)CR)(2) (L) (L')], (L=NH(3), L'=pyridine, quinoline, isoquinoline; L=L'=pyridine; R=H, CH(3), CH(2)OH, etc.) (with a focus on the contribution of the carboxylate leaving group to drug action) on growth and viability of A2780 human ovarian carcinoma cells as well as their putative mechanism(s) of cytotoxicity. The compounds, as a class, induce cell death through caspase-dependent apoptosis, with activation of both caspase 3 and caspase 9 and concomitant PARP cleavage. The trans-platinum compounds tested show induction of p53 as well as time dependent gammaH2AX induction, consistent with the promotion of DNA lesions. trans [Pt(O(2)CH)(2)(NH(3))(4-pic)] can be shown to promote significant DNA strand breaks and DNA interstrand cross-linking. The enhanced cytotoxicity of trans [Pt(O(2)CH)(2)(NH(3))(4-pic)] compared to its isostructural -O(2)CCH(3) and O(2)CCH(2)OH analogs may be a consequence of its accelerated cellular accumulation, increased hydrolytic activation, interstrand cross-linking and abortive efforts by the cell to repair the cross linked DNA. PMID- 17418818 TI - An experimental design approach for optimizing polycyclic aromatic hydrocarbon analysis in contaminated soil by pyrolyser-gas chromatography-mass spectrometry. AB - Pyrolyser-gas chromatography-mass spectrometry was used to analyze polycyclic aromatic hydrocarbons in contaminated soil without preliminary extraction. Experimental research methodology was used to obtain optimal performance of the system. After determination of the main factors (desorption time, Curie point temperature, carrier gas flow), modeling was done using a Box-Behnken matrix. Study of the response surface led to factor values that optimize the experimental response and achieve better chromatographic results. PMID- 17418820 TI - Echinostoma caproni: kinetics of IgM, IgA and IgG subclasses in the serum and intestine of experimentally infected rats and mice. AB - The kinetics of specific immunoglobulin M, A and IgG subclasses against Echinostoma caproni (Trematoda: Echinostomatidae) were analyzed in serum and intestinal fluid of two host species (Wistar rats and ICR mice) in which the course of the infection markedly differs. In rats, the worms were rapidly expelled, whereas E. caproni evokes in mice long-lasting infection. The pattern of antibody responses in both serum and intestinal samples was different in each host species. Serum responses in mice were characterized by significant increases of IgM, IgA, total IgG, IgG1 and IgG3, but not IgG2a. In contrast, serum responses in rats showed elevated levels of IgM, probably in relation to thymus independent antigens, and slight increases of total IgG, IgG1 and IgG2a. At the intestinal level, increases of IgM and IgA levels were observed in mice. In regard to IgG subclasses, increases in both IgG1 and IgG2a were detected. Later decreases to normal values in IgG2a were also detected. In rats, only increases in total IgG and IgG2a were found. According to our results the development of long-lasting E. caproni infections in mice appears to be associated with a dominance of Th2 responses at the systemic level and balanced Th1/Th2 responses at the local level, characterized by initial increases in IgG1 and IgG2a levels. In contrast, the worm expulsion appears to be related to increases in local IgG2a levels. PMID- 17418819 TI - Functional and molecular characterization of multiple K-Cl cotransporter isoforms in corneal epithelial cells. AB - The dependence of regulatory volume decrease (RVD) activity on potassium-chloride cotransporter (KCC) isoform expression was characterized in corneal epithelial cells (CEC). During exposure to a 50% hypotonic challenge, the RVD response was larger in SV40-immortalized human CEC (HCEC) than in SV40-immortalized rabbit CEC (RCEC). A KCC inhibitor-[(dihydroindenyl)oxy] alkanoic acid (DIOA)-blocked RVD more in HCEC than RCEC. Under isotonic conditions, N-ethylmaleimide (NEM) produced KCC activation and transient cell shrinkage. Both of these changes were greater in HCEC than in RCEC. Immunoblot analysis of HCEC, RCEC, primary human CEC (pHCEC), and primary bovine CEC (BCEC) plasma membrane enriched fractions revealed KCC1, KCC3, and KCC4 isoform expression, whereas KCC2 was undetectable. During a hypotonic challenge, KCC1 membrane content increased more rapidly in HCEC than in RCEC. Such a challenge induced a larger increase and more transient p44/42MAPK activation in HCEC than RCEC. On the other hand, HCEC and RCEC p38MAPK phosphorylation reached peak activations at 2.5 and 15 min, respectively. Only in HCEC, pharmacological manipulation of KCC activity modified the hypotonicity induced activation of p44/42MAPK, whereas p38MAPK phosphorylation was insensitive to such procedures in both cell lines. Larger increases in HCEC KCC1 membrane protein content correlated with their ability to undergo faster and more complete RVD. Furthermore, pharmacological activation of KCC increased p44/42MAPK phosphorylation in HCEC but not in RCEC, presumably a reflection of low KCC1 membrane expression in RCEC. These findings suggest that KCC1 plays a role in (i) maintaining isotonic steady-state cell volume homeostasis, (ii) recovery of isotonic cell volume after a hypotonic challenge through RVD, and (iii) regulating hypotonicity-induced activation of the p44/42MAPK signaling pathway required for cell proliferation. PMID- 17418821 TI - Up-regulation of the Ku heterodimer in Drosophila testicular cyst cells. AB - In Drosophila, developing germline cysts in testis are enveloped by two somatic cyst cells essential for germline development and male reproduction. The cyst cells continue development along with the germline. However, the mechanisms of somatic gene expression in testes are poorly understood. We report transcriptional up-regulation of the Ku heterodimer in cyst cells. The initial up regulation is independent of germline, and transcription is further augmented during spermatogenesis. Abundance of Ku in the cyst cell cytoplasm suggests the role for Ku subunits in the regulation of sperm individualization. PMID- 17418822 TI - What can Caenorhabditis elegans tell us about the nuclear envelope? AB - The nuclear envelope (NE) of the eukaryotic cell provides an essential barrier that separates the nuclear compartment from the cytoplasm. In addition, the NE is involved in essential functions such as nuclear stability, regulation of gene expression, centrosome separation and nuclear migration and positioning. In metazoa the NE breaks down and re-assembles around the segregated chromatids during each cell division. In this review we discuss the molecular constituents of the Caenorhabditis elegans NE and describe their role in post-mitotic NE re formation, as well as the usefulness of C. elegans as an in vivo system for analyzing NE dynamics. PMID- 17418823 TI - The Hansenula polymorpha peroxisomal targeting signal 1 receptor, Pex5p, functions as a tetramer. AB - We have studied Hansenula polymorpha Pex5p and Pex20p, peroxins involved in peroxisomal matrix protein import. In vitro binding experiments suggested that H. polymorpha Pex5p and Pex20p physically interact. We used single particle electron microscopy (EM) to analyze the structure of purified Pex5p and its possible association with Pex20p. Upon addition of Pex20p, a multimeric Pex20p complex was observed to be associated to the periphery of the Pex5p tetramer. In this Pex5p Pex20p complex, the conformation of tetrameric Pex5p had changed from a closed conformation with a diameter of 115A into an open conformation of 134A. EM also indicated that the Pex5p-Pex20p complex was capable to bind native, folded catalase, a peroxisomal PTS1 protein. This suggests that the Pex5p-Pex20p complex may be functional as receptor complex. PMID- 17418824 TI - Exposing cryptic antibacterial activity in Cyt1Ca from Bacillus thuringiensis israelensis by genetic manipulations. AB - In an attempt to endow Cyt1Ca with Cyt1Aa-like antibacterial activity, both derived from Bacillus thuringiensis subsp. israelensis, two amino acids were replaced, E117V and N125A, so as to raise the hydrophobicity of the corresponding region, considered to be the membrane-active motif. The clones obtained included multiple repeats of VIEVLKSLLGIALA, corresponding to head-to-tail polymerization of the primer, translated in frame with Cyt1Ca. These versions of Cyt1Ca caused instant arrest in biomass growth and decreased viability upon expression in Escherichia coli. Multiple insertions of the non-mutated motif VIEELKSLLGINLA into the polypeptide were also lethal. To expose toxicity of the latter motif in the original Cyt1Ca, cyt1Ca was appropriately truncated. PMID- 17418825 TI - Stress to endoplasmic reticulum of mouse osteoblasts induces apoptosis and transcriptional activation for bone remodeling. AB - ATF4 is an essential regulator in osteogenesis as well as in stress responses to the endoplasmic reticulum (ER). We addressed a question: Does ER stress to osteoblasts upregulate ATF4 expression? If so, do they exhibit ATF4-mediated bone remodeling or apoptosis? ER stress, induced by Thapsigargin and tunicamycin, elevated a phosphorylated form of eIF2alpha and ATF4, but the cellular fate depended on treatment duration. The treatment for 1h, for instance, activated Runx2, and type I collagen, while the treatment for 24h induced apoptosis. Our observations suggest that there is a threshold for ER stress and osteoblasts present a bi-phasic pattern of their fate. PMID- 17418827 TI - Update on the comparison of assisted reproduction outcomes between Europe and the USA: the 2002 data. AB - OBJECTIVE: In follow-up to an earlier study of in vitro fertilization (IVF) outcomes in Europe and the U.S. for the year 2001, the present study reports on 2002 outcomes in an attempt to determine diverging practice trends. DESIGN: Retrospective cohort comparisons. SETTING: Academically affiliated private fertility center. PATIENT(S): This study involved overall 324,238 IVF cycles in Europe and 115,538 cycles in the U.S. MAIN OUTCOME MEASURE(S): Pregnancy and delivery rates for fresh IVF cycles, frozen-thawed cycles, and oocyte donation cycles. RESULT(S): This study confirms continuing pregnancy outcome differences in all aspects of assisted reproduction to the benefit of the U.S., a much higher use of IVF procedures in Europe, and larger embryo transfer numbers and multiple pregnancy rates in the U.S., although both continents demonstrate an almost identical trend toward fewer embryos and lower multiple pregnancy rates. CONCLUSION(S): Universally lower pregnancy rates in Europe remain of concern, especially because the trend between 2001 and 2002 suggests a possible further widening of the gap. Initiatives that may further reduce European pregnancy rates, such as increasing emphasis on single-embryo transfers and legislative interventions, should therefore be introduced with caution. PMID- 17418826 TI - A proteasome for all occasions. AB - In the ubiquitin-proteasome system, substrates fated for destruction first acquire covalent modification by ubiquitin, and are subsequently destroyed by the proteasome. Traditionally, 26S proteasomes have been seen as largely uniform in their composition and functional capacity. Accordingly, cells can control proteasome abundance via transcriptional pathways that mediate concerted regulation of all known proteasome genes. However, recent evidence suggests that the proteasome is also subject to subunit-specific modes of regulation, which serve to alter proteasome function and may generate ensembles of compositionally distinct proteasomes. These modes of proteasome regulation provide varied means to adapt protein degradation pathways to changing conditions in the cell. PMID- 17418828 TI - Simple vaginal mold for use in the postoperative care of patients with a transverse vaginal septum. AB - Severe vaginal stenosis is a potentially disabling complication of transverse vaginal septum resection due to the constriction of the resulting circular scar. We describe a vaginal mold that can be easily created by an occupational therapist, and used as a long-term stent of the vagina in young girls. PMID- 17418829 TI - High incidence of complex chromosome abnormality in cleavage embryos from patients with repeated implantation failure. AB - OBJECTIVE: To analyze the chromosome abnormalities observed in the course of preimplantation aneuploidy screening using comparative genomic hybridization (CGH) on single blastomeres in relation to maternal age and previous IVF history. DESIGN: Retrospective analytical study. SETTING: A large IVF unit and the research laboratory of an associated clinical genetics unit. PATIENT(S): Twenty eight women referred for aneuploidy screening of cleavage embryos. INTERVENTION(S): Blastomere biopsy. MAIN OUTCOME MEASURE(S): The incidence of aneuploidy and complex abnormality in human cleavage embryos. RESULT(S): The incidence in embryos of aneuploidy for one to two chromosomes was significantly increased with advanced maternal age, but was independent of any history of recurrent implantation failure. In comparison, the incidence of complex chromosome abnormality (which involves three or more chromosomes) was independent of maternal age but significantly increased in embryos from patients with a history of recurrent implantation failure. CONCLUSION(S): The incidence of complex abnormality in healthy cleavage embryos is independent of maternal age but is increased in patients with a history of recurrent implantation failure. These results suggest that the pathology underlying complex abnormality is different from that resulting in aneuploidy of one to two chromosomes but particularly relevant to women with recurrent implantation failure. PMID- 17418830 TI - Korean ginseng induces spermatogenesis in rats through the activation of cAMP responsive element modulator (CREM). AB - The effect of Korean ginseng (ginseng) on spermatogenesis and cAMP-responsive element modulator (CREM) in rat testes was evaluated using sperm analysis, reverse transcription polymerase chain reaction, and Western blot analysis. The ginseng-treated rats exhibited significantly increased sperm count and motility with enhanced levels of CREM messenger RNA and protein. Ginseng appears to induce spermatogenesis via CREM activation in rat testes. PMID- 17418831 TI - Clinical-pregnancy outcome after vitrification of blastocysts produced from in vitro maturation cycles. AB - This study was conducted to report the clinical-pregnancy outcome after vitrification of the blastocysts produced from in vitro maturation cycles. The survival rate after thawing was 92.0% (92/100). The clinical-pregnancy and implantation rates were 43.8% and 23.6%, respectively. These results suggest that the blastocyst-stage embryos produced from in vitro maturation cycles can be safely cryopreserved through vitrification. PMID- 17418832 TI - Evaluation of effects of an oral contraceptive containing ethinylestradiol combined with drospirenone on adrenal steroidogenesis in hyperandrogenic women with polycystic ovary syndrome. AB - OBJECTIVE: To investigate whether the administration of an oral contraceptive containing the new antiandrogenic drospirenone is associated with reduced adrenal androgen synthesis in hyperandrogenic women with diagnosis of polycystic ovary syndrome. Drospirenone, an analogue of spironolactone and aldosterone antagonist, is a novel progestin under clinical development that is similar to the natural hormone progesterone, combining potent progestogenic with antimineralocorticoid and antiandrogenic activities. DESIGN: Prospective study. SETTING: Healthy volunteers in University Department of Obstetrics and Gynecology. PATIENT(S): Fifteen women ages 18 to 28 years with the diagnosis of polycystic ovary syndrome. INTERVENTION(S): Three months of contraceptive use (30 mcg ethinylestradiol, 3 mg drospirenone). MAIN OUTCOME MEASURE(S): An adrenocorticotropic hormone test was performed before and after the study. RESULT(S): Adrenal production of cortisol was unchanged after therapy with oral contraceptives. An interesting observation was reduced basal concentrations of androgens such as androstenedione, dehydroepiandrosterone sulfate, testosterone, and free testosterone during therapy. The ratios of the areas of substrates to products before and after oral contraceptive administration were compared for differences in 17alpha-hydroxylase (17-hydroxyprogesterone/progesterone) and 17,20-lyase (androstenedione/17-hydroxyprogesterone); activities were significantly reduced, indicating a reduction in the activities of these enzymes. CONCLUSION(S): The present results show for the first time that oral contraceptives containing drospirenone affect adrenal steroidogenesis by reducing synthesis and release of androgens in response to adrenocorticotropic hormone, leaving adrenal production of cortisol unchanged. PMID- 17418833 TI - Prognostic value of human chorionic gonadotropin changes after methotrexate injection for ectopic pregnancy. AB - Because early prediction of clinical outcome (one or more injections or surgery) of methotrexate treatment of ectopic pregnancy could ease the intensity of follow up and patient compliance required, we studied the relationship between the change in hCG levels after methotrexate injection and outcome in 129 consecutive patients. A 20% decline in hCG levels between days 1 and 4 during methotrexate treatment has a positive predictive value of 97%. PMID- 17418834 TI - Serum human chorionic gonadotropin level after ovulation triggering is influenced by the patient's body mass index and the number of larger follicles. AB - OBJECTIVE: To identify determinants of the serum concentration of hCG levels after triggering of ovulation with exogenous hCG during controlled ovarian stimulation cycles for in vitro fertilization with or without intracytoplasmic sperm injection. DESIGN: Retrospective cohort study. SETTING: University Medical Center. PATIENT(S): One hundred-fifteen women who underwent conventional in vitro fertilization/intracytoplasmic sperm injection cycles from March 2003 to March 2005. INTERVENTION(S): All patients underwent ovarian hyperstimulation with gonadotropins and GnRH-antagonist for pituitary downregulation. Patients were started on oral contraceptives 1 month before the stimulation. Gonadotropins were administered from stimulation day 1 until the day of the hCG trigger, and GnRH antagonist was added from the day when at least one follicle reached 14 mm in diameter and continued until hCG administration. The hCG was administered in 5,000-IU, 10,000-IU, or 15,000-IU doses on the day of ovulation triggering. MAIN OUTCOME MEASURE(S): We performed a stepwise multiple regression analysis to predict which variable would influence the serum concentration of hCG when measured the day after the administration of exogenous hCG. RESULT(S): Body mass index (kg/m2) and number of follicles >14 mm were the only determinants of the hCG concentration (cumulative R2 = 0.30; P<.001). Patient age, estradiol peak, number of oocytes retrieved, length of stimulation, and length of GnRH-antagonist administration in days were not associated with serum hCG levels. CONCLUSION(S): Knowing that the number of larger follicles and the patient's BMI are the major determinants of the hormone's clearance in the body can help in the hCG dose titration during ovarian stimulation. PMID- 17418835 TI - Serum total testosterone levels in a patient with late onset 21-hydroxylase deficiency and a twin gestation. AB - OBJECTIVE: To present serum androgen levels during pregnancy in a twin gestation complicated by maternal late onset 21-hydroxylase deficiency. DESIGN: Case report. SETTING: University teaching hospital reproductive endocrinology and infertility practice. PATIENT(S): A 27-year-old with nonclassic 21-hydroxylase deficiency and infertility, twin female fetuses, and elevated androgens. INTERVENTION(S): Steroid replacement. MAIN OUTCOME MEASURE(S): Serum T and 17 hydroxyprogesterone (17-OHP) levels. RESULT(S): Elevated androgen levels persisted throughout pregnancy in spite of aggressive steroid replacement. However, twin girls were born without any evidence of virilization. CONCLUSION(S): The changes associated with a twin gestation may result in excessive stimulation of androgens in mothers with nonclassic 21-hydroxylase deficiency. However, the increased placental aromatase provides protection. PMID- 17418836 TI - Cardiovascular risk factors are reduced with a low dose of acarbose in obese patients with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is defined by menstrual irregularity, hyperandrogenism, chronic anovulation, and enlarged ovaries with multiple follicles. Polycystic ovary syndrome is highly prevalent in women, affecting up to 10% of all women of reproductive age and reducing the possibility of spontaneous conception. In addition to altering reproductive function, PCOS has systemic implications, especially in the cardiovascular system. Cardiovascular risk (CVR) in PCOS patient increases because of insulin resistance, elevated androgen levels, and association with obesity. Those alterations promote cardiovascular risk factors, such as endothelial dysfunction, elevated homocysteine levels, left ventricular hypertrophy, and reduced high-density lipoprotein (HDL) cholesterol (1). PMID- 17418837 TI - Live birth after the transfer of human embryos developed from cryopreserved oocytes harvested before cancer treatment. AB - OBJECTIVE: To document the first successful fertility preservation with freezing mature oocytes for a cancer patient. DESIGN: Case report. SETTING: Private IVF center. PATIENT(S): A woman with Hodgkin's disease and her gestational carrier. INTERVENTION(S): Metaphase II oocytes cryopreserved before cancer treatments were thawed and fertilized for transfer to a gestational carrier. MAIN OUTCOME MEASURE(S): Post-thaw survival and pregnancy. RESULT(S): Ten of the 12 metaphase II oocytes survived the thawing process. Nine oocytes were fertilized after microinjection with her husband's sperm. All fertilized oocytes developed into good quality embryos and were transferred to a gestational carrier in three cycles. A biochemical pregnancy was achieved after the second transfer. A singleton pregnancy was achieved after the last transfer of three embryos, resulting in the birth of a healthy male. CONCLUSION(S): Our findings suggest that oocyte cryopreservation should be considered as a preferred strategy for the cancer patient who has time to cryopreserve oocytes before cancer treatment. PMID- 17418838 TI - Repeated pregnancies and live births after in vitro maturation treatment. AB - OBJECTIVE: To discuss and describe the successful treatment of women with primary infertility using in vitro maturation (IVM) of immature human oocytes resulting in repeated pregnancies and live births in the same individuals. DESIGN: Case report. SETTING: McGill Reproductive Center, Royal Victoria Hospital, McGill University. PATIENT(S): Six patients who underwent IVM treatment. INTERVENTION(S): In vitro maturation and fertilization of immature human oocytes. MAIN OUTCOME MEASURE(S): Clinical pregnancies and live births. RESULT(S): We report 11 singleton and 2 twin live births as a result of IVM treatment in six women with polycystic ovary syndrome. CONCLUSION(S): In vitro maturation of human oocytes is a novel treatment in the management of infertility. It is an effective method of conception, especially in women who have polycyctic ovaries. PMID- 17418839 TI - Comparison of the effect of single-dose and multiple-dose methotrexate therapy on tubal patency. AB - OBJECTIVE: To compare tubal patency by using hysterosalpingography (HSG) in women with unruptured ectopic tubal pregnancy who were treated with either single-dose or multiple-dose methotrexate (MTX) regimen. DESIGN: A case series with a retrospective statistical analysis. SETTING: Maternity and teaching hospital in Turkey. PATIENT(S): Sixty-one patients treated with MTX therapy for unruptured tubal ectopic pregnancy. INTERVENTION(S): Thirty-one patients received single dose and 30 multiple-dose MTX treatment. The HSG was performed 4 to 6 months after treatment to assess tubal patency. MAIN OUTCOME MEASURE(S): Ipsilateral and contralateral tubal obstruction rates. RESULT(S): Free passage through the ipsilateral tube was observed in 17 of 30 cases (56.7%) after multiple-dose, and 26 of 31 cases (83.9%) after single-dose MTX therapy. Patency of the contralateral tube was higher after single-dose than multiple-dose MTX treatment, although the value was not statistically significant. Binary logistic regression analysis was used to evaluate the parameters of age, gravida, parity, initial titers of beta human chorionic gonadotropin, size of the adnexal mass, and MTX regimen and their correlation with the HSG results after the clinical treatment for unruptured ectopic pregnancy. Only the type of MTX regimen was found to be used as a predictor of ipsilateral tubal obstruction. CONCLUSION(S): In terms of ipsilateral tubal obstruction, multiple-dose MTX therapy appears to have a greater negative effect on tubal patency than single-dose therapy. PMID- 17418840 TI - Obesity and poor reproductive outcome: the potential role of the endometrium. AB - OBJECTIVE: To analyze the potential role of extraovarian factors such as endometrium in the reproductive outcome of obese patients. DESIGN: Retrospective study. SETTING: University-affiliated infertility clinic, between January 2001 and July 2005. PATIENT(S): Women undergoing a total of 2656 first ovum donation (OD) cycles with good quality embryos. INTERVENTION(S): The oocyte donors underwent controlled ovarian hyperstimulation, and recipients received a well established hormonal replacement therapy for endometrial preparation. In vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) was performed according to semen characteristics. All first cycles (n = 2656) of ovum donation used good quality embryos and were divided into groups according to body mass index: <20 kg/m(2) (n = 471), 20 to 24.9 kg/m(2) (n = 1613), 25 to 29.9 kg/m(2) (n = 450), and > or =30 kg/m(2) (n = 122). MAIN OUTCOME MEASURE(S): The recipient body mass index groups were compared regarding IVF outcome. RESULT(S): The rates of implantation, pregnancy, miscarriage, and ongoing pregnancy were similar among the body mass index groups, although there was a negative trend when body mass index increased. Ongoing pregnancy rates per cycle were poorer in the overweight and obese groups than in the underweight and normal groups. In addition, women under 25 kg/m(2) presented an ongoing pregnancy rate per cycle of 45.5%, compared with 38.3% for those with > or =25 kg/m(2). CONCLUSION(S): Excess weight exerts an extraovarian detrimental effect. The role of the endometrium or its environment seems to be subtle but should be taken into account. PMID- 17418841 TI - The effect of 3,5,3'-triiodothyronine supplementation on zebrafish (Danio rerio) embryonic development and expression of iodothyronine deiodinases and thyroid hormone receptors. AB - The importance of thyroid hormones (TH) for embryonic development has long been shown in many vertebrates, but is not yet established in pre-hatch teleost models despite the presence of TH, TH receptors and iodothyronine deiodinases. Lack of data about the dynamics of TH metabolism in embryonic stages of fish does not allow to speculate about the involvement and/or role of TH in fish embryonic development. We therefore set up an experiment to examine the effect of 3,5,3' triiodothyronine (T(3)) on zebrafish (Danio rerio) embryonic development and on the expression of some thyroid hormone-regulated genes as measured by real-time PCR. Maternally derived thyroxine (T(4)) and T(3) were detected throughout embryonic development and total levels remained stable. Thyroid hormone receptor (TR) alpha and beta mRNA were found to be present in early embryos. After an initial fall, TRalpha mRNA levels in the control group increased gradually from 12h post fertilization (HPF) onwards. TRbeta mRNA levels of control embryos were relatively stable during embryonic development, but increased around the hatching period. We also quantified type I (D1) and type II (D2) deiodinase mRNA expression in zebrafish embryos. D1 mRNA levels in the control group gradually increased during development while D2 levels were kept at a low and stable level until hatching. At 75 HPF, a fivefold increase of D2 expression was observed compared to embryonic levels. Exogenous T(3) added to the water (5nM) was taken up by the embryos, causing increased pigmentation and accelerated hatching. T(3) treatment significantly up regulated TRalpha mRNA levels at 48 HPF, while D2 mRNA was significantly down regulated at 75 HPF. Neither TRbeta nor D1 mRNA levels seemed responsive to the treatment. Taken together, these data show that during embryonic development zebrafish already have the necessary regulatory machinery for TH activation and signaling, and that T(3) treatment at that stage indeed influence embryonic development. PMID- 17418842 TI - cDNA cloning of proopiomelanocortin (POMC) and mass spectrometric identification of POMC-derived peptides from snake and alligator pituitaries. AB - Proopiomelanocortin (POMC) is the precursor of melanocyte-stimulating hormone (MSH) and beta-endorphin, and is suggested to have evolved by the insertion and deletion of ancestral MSH segments. Here, the primary structure of POMC was determined with cDNA cloning of brown tree snakes of Squamata and American alligators of Crocodylia to show an overview of the molecular evolution of POMC in reptiles. Snake and alligator POMCs are composed of alpha-, beta-, and gamma MSH segments and a single beta-END segment as in other tetrapods; however, the gamma-MSH segment in snake POMC has a mutation in the essential sequence from His Phe-Arg-Trp to His-(d)-(d)-Arg, in which (d) means deletion. It is conceivable that the ancestry of snake gamma-MSH had weak functional constraint and lacked biological significance during evolution. Phylogenetic analyses using the neighbor-joining method show that snake prePOMC is most diverged, and alligator prePOMC is most conserved in reptilian POMCs while it shows the highest sequence identity with ostrich prePOMC. These relationships are comparable to those observed in mitochondrial DNA. On the other hand, analyses of the pituitary with mass spectrometry revealed several peptides by post-translational processing as predicted by the locations of processing sites consisting of basic amino acid residues in snake and alligator POMCs. Remarkably, the monobasic site at the N terminal side of the snake beta-MSH is suggested to act as a processing site. Thus, the study shows the divergence of snake POMC such as the critical mutation of gamma-MSH and high conservation of hormone organization of alligator POMC. PMID- 17418843 TI - Development of real-time RT-PCR assays for eel gonadotropins and their application to the comparison of in vivo and in vitro effects of sex steroids. AB - Gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), are key factors in the brain-pituitary-gonad axis and understanding their regulation remains essential for future management of eel reproduction. In this regard, we developed quantitative real-time RT-PCR (qrtRT-PCR) assays for the expression of European eel LHbeta, FSHbeta and GPalpha subunits, using the Light Cycler system. The qrtRT-PCR was adapted to permit detection of the three gonadotropin subunit mRNAs in individual pituitaries and in dispersed pituitary cells. The validated assays were applied to investigate the effects of sex steroids (estrogens and androgens) on gonadotropin subunit expression, in vivo in steroid-injected eels, and in vitro by steroid treatments of primary cultures of eel pituitary cells. In vivo, a stimulation of LHbeta mRNA was observed after estradiol (E2) treatments, while testosterone (T) or the non-aromatizable androgen dihydrotestosterone (DHT) had no effect. Concerning FSHbeta expression, slight but non-significant decreases were observed after both E2 and androgen treatments. Different results were obtained in vitro: E2 induced an increase in FSHbeta mRNA levels but had no effect on LHbeta expression. In contrast, androgens (T and DHT) stimulated LHbeta expression while no significant variation was observed on FSHbeta mRNA levels following androgen treatment. Concerning the GPalpha mRNA, no significant effect of sexual steroids was observed in vivo or in vitro. This demonstrated specific direct actions of steroids on gonadotropin subunit expression. The differences observed between in vivo and in vitro experiments may be explained by the involvement of cerebral control, including GnRH and dopamine neurons, and their specific regulation by sex steroids. The data indicate that sex steroid feedbacks on gonadotropins are exerted via multiple pathways, indirectly at the brain level and directly on pituitary gonadotrope cells. PMID- 17418844 TI - Role of nerve growth factor and its receptor in the morphogenesis of neural tube in early chick embryo. AB - Expression of p75 nerve growth factor receptor (p75(NTR)) in the early neurogenesis of chick embryo showed that nerve growth factor receptor (NGFR) is localized in presumptive neuroectoblast and endoblast in the chick gastrula but not in the mesoblast. By stages 9, 10, and 11, NGFR positive cells were located distinctly in the region where the neural folds converge, meet and fuse. NGFR expression was also seen in developing notochord and somites, wherein the reaction was localized on the cell surfaces. Strong p75(NTR) reaction was seen on the roof of the neural tube where it detaches from the head ectoderm by stage 12. The study revealed that p75(NTR) is co-expressed with NGF in the same developmental stage(s) and in areas, where cell death occurs during neuronal development. Further, when the endogenous levels of NGF signaling were blocked by anti-NGF antibody, abnormalities were observed at the anterior end of the neural tube formation. As a result, embryos showed open neural tubes and a few were bent on one side of the body axis. In a small proportion of embryos, diffused somites were observed. The findings supports and confirms our previous study that NGF signaling plays a significant role in the shaping of neural tube in chick embryos through p75(NTR)-NGF receptor. PMID- 17418845 TI - Reproductive stage-related effects of salmon GnRH and sex steroid hormones on expression of genes encoding fushi tarazu factor 1 homolog and estrogen receptor alpha in masu salmon pituitary cells in vitro. AB - Expression of genes encoding gonadotropin (GTH) subunits in the salmon pituitary was regulated by salmon gonadotropin-releasing hormone (sGnRH) and sex steroid hormones in a reproductive stage-dependent manner, probably through DNA-binding transcription factors. Direct effects of these hormones on expression of genes encoding salmon fushi tarazu factor 1 homolog (sFF1-I) and estrogen receptor alpha (ERalpha) were therefore examined by use of primary pituitary cell cultures of masu salmon at different reproductive stages. Pituitaries were collected in March (before initiation of gonadal maturation), in May (early maturing), in July (late maturing), and in September (spawning period). Amounts of sFF1-I and ERalpha mRNAs in the pituitary cells were determined by real-time polymerase chain reactions after a treatment with sGnRH, estradiol-17beta (E2), testosterone (T) or 11-ketotestosterone (11KT). The amounts of sFF1-I mRNA were elevated by E2 in the males, and by sGnRH and T in the females before initiation of gonadal maturation and at the early maturing stage. The amounts of ERalpha mRNA in the early maturing females were elevated by sGnRH. Effects of sGnRH were not significant at the late maturing and spawning stages. The amounts of ERalpha mRNA in the spawning males were halved by 11KT and E2, and those of sFF1-I and ERalpha mRNAs in the late maturing females were decreased by T and 11KT. These results indicated that responsiveness of sFF1-I and ERalpha genes to sGnRH and sex steroid hormones is seasonally variable in relation to reproductive stages. Expression of sFF1 and ERalpha genes should be stimulated at the early stages of gonadal maturation prior to increases in the amounts of GTH subunit mRNAs, while attenuated after the late maturing period when stored amounts of GTH subunit mRNAs reached near the maximum. PMID- 17418846 TI - Molecular cloning of PEPCK and stress response of black porgy (Acanthopagrus schlegeli) to increased temperature in freshwater and seawater. AB - Stress responses to increased temperature in black porgy reared in freshwater (FBP) and seawater (SBP) were examined via endocrinological and blood physiological methods. A rise in temperature increased plasma cortisol levels, which were significantly higher in FBP compared to SBP. The stimulated expression of phosphoenolpyruvate carboxykinase (PEPCK) mRNA in liver might result from the high cortisol level, and this explains the observed higher plasma glucose levels in FBP versus SBP. Full-length cDNA sequence for PEPCK was determined by 3' and 5' RACE procedures. PEPCK cDNA clone was found to contain 2563 nucleotides including an open reading frame that encodes 624 amino acids. While aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of FBP increased with temperature, there was no change in SBP. In FBP, T(3) were 2.3+/-0.3 ng/ml at 20 degrees C and significantly decreased to 1.0+/-0.3 ng/ml at 30 degrees C. On the other hand, in SBP, it were 3.1+/-0.5 ng/ml at 20 degrees C but significantly increased to 5.2+/-0.4 ng/ml at 30 degrees C. When comparing osmolality at the temperature of 30 degrees C and of 20 degrees C, the difference was found to be greater for FBP than SBP. Accordingly, the results suggest that FBP suffers greater stress than SBP with increased temperature, and provide stress responses and osmoregulatory abilities against stressors in black porgy that could differ depending on salinities. PMID- 17418847 TI - Embedded model control: application to web winding. AB - Embedded Model Control (EMC) methodology is applied to web-winding control problems. These require time-varying Embedded Model and adaptive control law, because of a variable winder radius; disturbance estimation and rejection, because of friction and variable web speed and acceleration; sensorless reel radius estimation; and robust stability and performance in the presence of web and gear vibrations. The goal is to solve these key issues in the framework of EMC, which amounts to (i) designing a time-varying Control Law capable of disturbance rejection, (ii) designing Noise Estimators capable of keeping live and accurate disturbance and radius estimates, and (iii) tuning their eigenvalues to guarantee stability and performance. Tuning takes advantage of an 'error loop' formulation, which explicitly relates neglected dynamics to eigenvalues, stability and performance. Simulated runs from a fine winder model are compared to published results. PMID- 17418848 TI - Health visiting assessment processes under scrutiny: a case study of knowledge use during family health needs assessments. AB - BACKGROUND: Accurate assessment of family health need is a core health visiting skill, requiring considerable knowledge and expertise. To date, there has been only limited empirical examination of the types of knowledge and ways of knowing which are likely to influence health visiting practice during family health needs assessments. OBJECTIVES: This paper will present a detailed analysis of health visiting assessment processes and will explicate some of the many elements associated with the processes of identifying and assessing family health needs. DESIGN: An in-depth case study was undertaken to explore health visiting practice across three study sites. The focus of interest was to attempt to understand the factors that may influence a health visitor in making a professional judgement to offer a family extra support. SETTINGS: The study was conducted in three community Trust case sites in England, UK. METHODS AND PARTICIPANTS: The study was informed by a constructivist methodology. Data collection took place during 56 observed home visits to families receiving increased health visiting support and intervention. Following the home visits separate in-depth interviews were undertaken with the health visitors and the clients. RESULTS/CONCLUSIONS: This paper will explicate some of the many elements associated with the processes of identifying and assessing family health needs. It endeavours to unravel some of the complexity and intricacies of these processes and provide insights into health visitors' practical 'know-how'. PMID- 17418849 TI - The administration of estrogens, combined with anti-androgens, has beneficial effects on the hormonal features and asymmetric dimethyl-arginine levels, in women with the polycystic ovary syndrome. AB - The present study was designed in order to: (a) compare ET-1 and ADMA levels, between women with PCOS (n=106) and healthy controls (n=30); (b) determine the effects of treatment with estrogens and anti-androgens on the hormonal features of PCOS, insulin resistance, ET-1 and ADMA levels. Women with PCOS were randomized in five therapeutic protocols: (I) 17beta-estradiol+cyproterone acetate 50mg; (II) conjugated estrogen+CA 50 mg; (III) ethinyl estradiole+CA 2mg; (IV) EE+CA 52 mg; (V) EE+desogestrel. In all women, gonadotropin, PRL, androgen, SHBG, insulin, glucose, ET-1 and ADMA levels were determined; in women with PCOS, testosterone, SHBG, ET-1 and ADMA levels were measured again after 3, 6, 12 months of treatment and insulin and glucose levels after 12 months. ET-1 and ADMA concentrations were higher in women with PCOS, and they were positively correlated with each other. ADMA levels were decreased and IR was increased with treatment. Treatment with synthetic estrogens (EE) resulted in a more pronounced increase in SHBG and a more pronounced decrease in FAI, compared to natural estrogens. Conclusively, PCOS is associated with endothelial dysfunction, which is ameliorated by the administration of estrogens and anti-androgens, independent of IR. PMID- 17418850 TI - Effect of geometrical imperfections in confined compression tests on parameter evaluation of hydrated soft tissues. AB - Experimental tests, such as the confined and unconfined compression and the indentation tests, are traditionally used to determine the poroelastic properties of hydrated soft tissues (HSTs). The purpose of this study was to quantitatively evaluate the reliability of H(A) and K values as identified from experimental confined test data, estimating the errors that could occur in several situations with more realistic sample geometry and boundary conditions. Finite element models of the step-wise stress-relaxation confined compression tests on HSTs were developed including geometrical imperfections of the sample and the presence of a gap between the piston and the confining chamber. The errors occurring when H(A) and K were estimated by means of the analytical solution of the 1-D confined compression problem were assessed. Results of the analysis indicate that errors in the parameter estimation due to geometrical inaccuracies of the sample can be eliminated by applying a 5% strain pre-compression to the sample. Gap errors are negligible for H(A), can reach 20% for K, and cannot be eliminated by a pre compression of the sample. PMID- 17418851 TI - One-dimensional and comprehensive two-dimensional gas chromatography coupled to soft photo ionization time-of-flight mass spectrometry: a two- and three dimensional separation approach. AB - Soft laser photo-ionization mass spectrometry is presented as a separation dimension hyphenated with gas chromatographic techniques. Single photon ionization (SPI) is a universal soft ionization method which ionizes organic molecules with an ionization potential below 10.5 eV if 118 nm laser radiation is used. The inherently soft ionization of photo ionization techniques can further be utilized together with gas chromatography as a comprehensive two-dimensional separation method (GC x MS), using the GC retention time as first separation dimension and the molecular mass as second separation dimension. Some GC x MS chromatograms of diesel petroleum samples using SPI are presented and discussed. Finally, it is demonstrated that the coupling of soft SPI mass spectrometry with comprehensive two-dimensional gas chromatography (GC x GC) provides a three dimensional separation technique (GC x GC x SPI-MS). PMID- 17418852 TI - trans-(1R,2R)-Diaminocyclohexane-functionalized mesoporous organosilica spheres as chiral stationary phase. AB - The bifunctionalized mesoporous organosilica spheres with trans-(1R,2R) diaminocyclohexane (DACH) in the pore were synthesized and their application as chiral stationary phase in high-performance liquid chromatography (HPLC) was demonstrated. Bifunctionalized mesoporous organosilica spheres with narrow particle size distribution of 5-7 microm were prepared by the co-condensation of N-[3-(triethoxysilyl)propyl]-trans-(1R,2R)-diaminocyclohexane (M(propyl)) with 1,2-bis(trimethoxysilyl)ethane (BTME) in basic medium using octadecyltrimethylammonium chloride (C(18)TACl) as structural directing agent and ethanol as co-solvent. The morphologies of the bifunctionalized mesoporous organosilicas were sensitive to the molar fraction of M(propyl). At higher M(propyl)/BTME molar ratio, the bifunctionalized organosilica spheres with ordered mesoporous structure can be formed in a wide range of synthetic conditions. When the molar percent of M(propyl)/(M(propyl)+BTME) is less than 20, the formation of spheres could be hardly observed. The structural order of the materials mainly depends on the base concentration and M(propyl)/BTME molar ratio in the initial sol mixture. A column packed with the bifunctionalized mesoporous organosilica spheres exhibits higher selectivity and resolution for racemic amino acids than that packed with DACH-SiO(2) prepared by conventional post-synthesis grafting method. PMID- 17418853 TI - Investigation of pore diffusion hindrance of monoclonal antibody in hydrophobic interaction chromatography using confocal laser scanning microscopy. AB - In this article, hindrance of intraparticle mass transfer during the adsorption of a monoclonal antibody (mAb) on Butyl Sepharose 4FF was investigated under different conditions. In addition to common fluid phase measurements, confocal laser scanning microscopy (CLSM) was applied to evaluate the respective intraparticle concentration profiles. In order to ensure that the observed intraparticle protein distributions are not disturbed by artefacts of CLSM, microscopic data are carefully analysed considering signal attenuation and competitive adsorption between labelled and native species. Using this setup, lower protein concentration in the inner particle region was observed even after long equilibration times in protein solution. Since the observed phenomenon showed a dependency on the amount of adsorbed protein, we assumed that the intraparticle diffusion was hindered by the adsorbed protein molecules. We propose hypotheses on the diffusion hindrance, and compare the experimental results with model-based simulations of single particles that include novel terms for the description of hindered diffusion. PMID- 17418854 TI - Automation of solid-phase microextraction on a 96-well plate format. AB - Studies have been performed assessing the feasibility and characterizing the automation of solid-phase microextraction (SPME) on a multi-well plate format. Four polycyclic aromatic hydrocarbons (PAHs), naphthalene, fluorene, anthracene and fluoranthene, were chosen as test analytes to demonstrate the technique due to their favorable partition coefficients, K(fw), between polydimethylsiloxane (PDMS) extraction phases and water. Four different PDMS configurations were investigated regarding their suitability. These included (i) a PDMS membrane; (ii) a multi-fiber device containing lengths of PDMS-coated flexible wire; (iii) a stainless steel pin covered with silicone hollow fiber membrane and (iv) commercial PDMS-coated flexible metal fiber assemblies. Of these configurations, the stainless steel pin covered with silicone tubing was chosen as a robust alternative. An array of 96 SPME devices that can be placed simultaneously into a 96-well plate was constructed to demonstrate the high-throughput potential when performing multiple microextractions in parallel. Different agitation methods were assessed including magnetic stirring, sonication, and orbital shaking at different speeds. Orbital shaking whilst holding the SPME device in a stationary position provided the optimum agitation conditions for liquid SPME. Once the analytes had been extracted, desorption of the analytes into an appropriate solvent was investigated. Liquid-phase SPME and solvent desorption on the multi well plate format is shown to be a viable alternative for automated high throughput SPME analysis compatible with both gas- and liquid-chromatography platforms. PMID- 17418855 TI - Determination of four lignans in Phyllanthus niruri L. by a simple high performance liquid chromatography method with fluorescence detection. AB - A new and simple analytical method using HPLC with fluorescence detection was developed for the simultaneous determination of four lignans (phyllanthin, hypophyllanthin, phyltetralin and niranthin) in Phyllanthus niruri L. plant samples. Optimal separation was achieved with an isocratic mobile phase consisting of acetonitrile-water (55:45 v/v). The method recorded limits of detection (S/N=5) for phyllanthin at 0.61 ng/mL, hypophyllanthin at 6.02 ng/mL, phyltetralin at 0.61 ng/mL and niranthin at 1.22 ng/mL, being 80, 8, 80 and 40 times, respectively, lower when compared with those derived using HPLC-UV detection. The limits of quantification (S/N=12) were 4.88 ng/mL for phyllanthin and phyltetralin, 9.76 ng/mL for niranthin and 24.4 ng/mL for hypophyllanthin showing 40, 8 and 20 times, respectively, lower than those from the UV detection method. The within-day and between-day accuracy for the four lignans were between 98.1% and 102.9% while their precision values were below 2.2%. The mean recovery was between 92.5% and 110.1%. The method was then successfully applied for the quantification of lignans in P. niruri plant samples. The highest amount of lignans was found in the leaves followed by fruits, branches and stem, whilst the roots have the least amount of lignans. PMID- 17418856 TI - Thermal stability of octadecyltrimethylammonium bromide modified montmorillonite organoclay. AB - Organoclays are significant for providing a mechanism for the adsorption of organic molecules from potable water. As such their thermal stability is important. A combination of thermogravimetric analysis and infrared emission spectroscopy was used to determine this stability. Infrared emission spectroscopy (IES) was used to investigate the changes in the structure and surface characteristics of water and surfactant molecules in montmorillonite, octadecyltrimethylammonium bromide and organoclays prepared with the surfactant octadecyltrimethylammonium bromide with different surfactant loadings. These spectra collected at different temperatures give support to the results obtained from the thermal analysis and also provide additional evidence for the dehydration which is difficult to obtain by normal thermoanalytical techniques. The spectra provide information on the conformation of the surfactant molecules in the clay layers and the thermal decomposition of the organoclays. Infrared emission spectroscopy proved to be a useful tool for the study of the thermal stability of the organoclays. PMID- 17418857 TI - Synthesis, characterization, and visible light activity of new nanoparticle photocatalysts based on silver, carbon, and sulfur-doped TiO2. AB - New nanoparticle photocatalysts based on silver, carbon, and sulfur-doped TiO2 having only the homogeneous anatase crystalline phase and high surface area were successfully synthesized by a modified sol-gel route. The catalysts were characterized by EDX, XRD, BET, UV-vis, IR, and Raman spectroscopy. The effects of the experimental parameters on the visible light reactivity of the catalysts were evaluated for the photodegradation of gaseous acetaldehyde as a model indoor pollutant. The activity results show that the silver(I) ion, Ag(+), doping significantly promotes the visible light reactivities of carbon and sulfur-doped TiO2 catalysts without any phase transformation from anatase to rutile. Moreover, Ag/(C, S)-TiO2 photocatalysts degrade acetaldehyde 10 times faster in visible light and 3 times faster in UV light illuminations than the accredited photocatalyst P25-TiO2. The commendable visible photoactivities of Ag/(C, S)-TiO2 new nanoparticle photocatalysts are predominantly attributable to an improvement in anatase crystallinity, high surface area, low band gap and nature of precursor materials used. PMID- 17418858 TI - Investigations of drop impact on dry walls with a lattice-Boltzmann model. AB - In this work, axisymmetric computations of drop impingement on dry walls are presented. The two-phase model employed is an axisymmetric lattice-Boltzmann model. Computations are performed in the parametric range of Weber number We of 7 to 8770, Ohnesorge number Oh of 0.02 to 0.137, and drop-wall equilibrium contact angle theta(eq) of 35 degrees to 150 degrees . Deposition and rebound outcomes are reported. In deposition, the different stages of drop evolution including spread, recoil and equilibration are reproduced and studied. Comparisons made with experimentally reported data of temporal evolution of the spread factor and the dynamic evolution of the contact angle show good agreement. Rebound is observed on non-wetting surfaces. The transition between deposition and rebound is shown to be influenced by the impact We, Oh, and advancing and receding static contact angles. Apart from impingement outcomes, the influence of We and Oh on the dynamic contact angle is investigated. PMID- 17418859 TI - Relationship between N,N-dialkyl chitosan monolayer and corresponding vesicle. AB - The properties of N,N-dialkyl chitosan monolayers and corresponding vesicles have been studied by LB technique and drug-release experiment. With increasing molecular weight of chitosan backbone and/or length of alkyl chain, both the compressibility and collapse pressure of N,N-dialkyl chitosan monolayer are enhanced. The experiments on drug-release behavior of N,N-dialkyl chitosan vesicles show that the drug-release rate and the equilibrium drug-release ratio are decreased with increasing the compressibility of corresponding monolayer. It is worth noticing that there is a linear relationship between the compressibility of N,N-dialkyl chitosan monolayers and the equilibrium drug-release ratio of the vesicles. PMID- 17418860 TI - Dissecting choral speech: properties of the accompanist critical to stuttering reduction. AB - The effects of choral speech and altered auditory feedback (AAF) on stuttering frequency were compared to identify those properties of choral speech that make it a more effective condition for stuttering reduction. Seventeen adults who stutter (AWS) participated in an experiment consisting of special choral speech conditions that were manipulated to selectively eliminate specific differences between choral speech and AAF. Consistent with previous findings, results showed that both choral speech and AAF reduced stuttering compared to solo reading. Although reductions under AAF were substantial, they were less dramatic than those for choral speech. Stuttering reduction for choral speech was highly robust even when the accompanist's voice temporally lagged that of the AWS, when there was no opportunity for dynamic interplay between the AWS and accompanist, and when the accompanist was replaced by the AWS's own voice, all of which approximate specific features of AAF. Choral speech was also highly effective in reducing stuttering across changes in speech rate and for both familiar and unfamiliar passages. We concluded that differences in properties between choral speech and AAF other than those that were manipulated in this experiment must account for differences in stuttering reduction. LEARNING OUTCOMES: The reader will be able to (1) describe differences in stuttering reduction associated with altered auditory feedback compared to choral speech conditions and (2) describe differences between delivery of a second voice signal as an altered rendition of the speakers own voice (altered auditory feedback) and alterations in the voice of an accompanist (choral speech). PMID- 17418861 TI - Mast cell-derived cathepsin g: a possible role in the adverse remodeling of the failing human heart. AB - BACKGROUND: The role of cardiac mast cells (MCs) in the progression to heart failure has recently become increasingly evident. Cathepsin g is a neutrophil- and mast cell-derived protease, which can convert angiotensin I to angiotensin II and thereby activate the TGF-beta pathway, resulting in myocyte necrosis, hypertrophy, and increased fibrosis. This study focuses on mast cell-derived cathepsin g in the human heart during heart failure and following mechanical unloading by means of heart-assist devices (LVADs). MATERIALS AND METHODS: Myocardial tissue was obtained from 10 patients with end-stage cardiomyopathy at the time of LVAD implantation (pre-LVAD) and following orthotopic heart transplantation (post-LAVD). In addition, biopsies of four normal hearts served as a control group. Paraffin-embedded sections were dual stained for cathepsin g and tryptase, a known marker for mast cells, using standard immunohistochemistry protocols. Total cathepsin g positive mast cells were counted. RESULTS: No cathepsin g positive MCs were found in normal hearts. However, we found evidence for cathepsin g in cardiac MCs in heart failure tissues (pre-LVAD). During heart failure, 46% of total MCs were cathepsin g positive as compared to after mechanical unloading, where only 11% of total MCs were cathepsin g positive (P<0.001). CONCLUSION: Heart failure causes an increase of myocardial MCs. We have provided evidence that cathepsin g positive MCs accumulate during heart failure and their total percentage decreases after ventricular unloading. This coincides with the decrease in myocyte necrosis, hypertrophy, and fibrosis. Thus, cathepsin g may play a role in the progression to heart failure by activating angiotensin II, leading to detrimental effects on the heart. PMID- 17418862 TI - Trends in mammography and clinical breast examination: a population-based study. AB - BACKGROUND: There are well-accepted clinical guidelines for early detection of breast cancer through mammography and clinical breast examination. The purpose of this study was to determine trends in the utilization of these techniques over the past 5 years. METHODS: The National Health Interview Survey is a population based computer survey of noninstitutionalized civilian Americans conducted annually by the National Center for Health Statistics, Centers for Disease Control and Prevention. We evaluated the reported use of mammography and clinical breast examination in women 40 years of age and older between 2000 and 2005. Data were evaluated using SAS and SAS-callable SUDAAN software. RESULTS: Surveyed were 10,994 and 11,128 women over age 40 in 2000 and 2005, respectively. Between these years, there was a decline in women >or=40 years old who reported ever having a mammogram (80.79% versus 79.52%, P<0.0001) and in those who had a mammogram within the preceding 2 years (87.98% versus 86.30%, P=0.0040). In addition, there was a drop in women reporting ever having a clinical breast exam (82.12% versus 75.91%, P<0.0001) and in those reporting having had a clinical breast exam in the 2 years prior to the survey (89.24% versus 87.63%, P=0.0012). Similar results were found in the population aged 50-69. CONCLUSIONS: Despite well-established clinical guidelines for early detection of breast cancer, there has been a decline in the rates of annual mammography and clinical breast exam in women over the age of 40 over the past 5 years. PMID- 17418863 TI - Evaluation of postoperative peritoneal adhesion formation following perioperative nicotine administration. AB - PURPOSE: The primary step in postoperative peritoneal adhesion formation involves the exudation of fibrin through permeable mesenteric capillaries. Nicotine, the most potent constituent of cigarette smoke, augments the release of vascular endothelial growth factor (VEGF), which increases vascular permeability. The objective of this study was to evaluate the effect of nicotine on postlaparotomy abdominal adhesions. METHODS: CD-1 mice were randomized to receive, ad libitum, either water alone (as 2% sucrose solution) or water mixed with 100 microg/mL nicotine (to obtain blood levels equivalent to a 1 pack/day smoker) (n = 32 mice per group). After 1 wk, all mice underwent laparotomy to induce adhesion formation using the same experimental method (consisting of laparotomy with cecal abrasion to achieve serosal punctuate hemorrhage). Fourteen days later, all animals were sacrificed for adhesion grading (0 to 3) by a blinded observer. Additionally, peritoneal lavage fluid from separate mice receiving either water or nicotine water before their surgery or else undergoing sham laparotomy was retrieved at 2, 6, and 24 h postoperatively (4 mice per group/time point) and assessed for VEGF concentrations. RESULTS: The nicotine treated group had a mean +/- SEM adhesion score of 2.4 +/- 0.2, which was significantly greater than that of the control group (1.2 +/- 0.2) (P < 0.05, Mann Whitney U test.). This correlated with a significant increase in peritoneal VEGF levels occurring at 6 and 24 h in the nicotine group. CONCLUSION: Peritoneal adhesion formation after surgery is increased in mice receiving nicotine perioperatively. This effect may be mediated through a direct augmentation of peritoneal VEGF release by nicotine with a subsequent increase in mesenteric endothelial permeability. These findings could have significant implications for smokers undergoing intra-abdominal surgical procedures. PMID- 17418864 TI - Cyclooxygenase-2 expression induces genomic instability in MCF10A breast epithelial cells. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) is induced in many breast cancers and COX-2 expression correlates with a worse outcome in the clinic. We hypothesized that the induction of genomic instability is a major mechanism through which COX-2 contributes to breast cancer progression. METHODS: We transfected a normal immortalized breast epithelial cell line of Basal B subtype, MCF10A, with the pSG5-COX-2 vector and established the stably transfected cell line MCF10A/COX-2. We analyzed the genomic instability phenotype by chromosomal analysis of metaphase-arrested MCF10A and MCF10A/COX-2 cells after Giemsa staining. Groups were compared using chi(2) tests. To investigate the DNA damage checkpoint signaling, we analyzed the phosphorylation status of CHK1 protein with a phospho specific antibody. RESULTS: Cytogenetic analysis of early passage transfected cells showed that COX-2 expression increased genomic instability compared with the MCF10A cells transfected with a luciferase vector alone. COX-2 overexpression was associated with a significant increase in chromosomal aberrations (fusions, breaks, and tetraploidy). There was a statistically significant increase in the number of polyploid cells in the COX-2 transfected cells versus the control (P=0.004). We also found that an inhibitory CHK1 phosphorylation at Ser-280 was dramatically increased upon COX-2 overexpression in MCF10A cells, thus explaining the mechanism of inactivation of an important cell cycle checkpoint. Further analysis of the MCF10A/COX-2 cells showed that these cells have acquired a premalignant phenotype characterized by a morphological transformation, a resistance to anoikis, a reduced requirement of epidermal growth factor for growth in culture, but their inability to establish tumors in a nude mouse model of malignancy. CONCLUSION: We found that COX-2 expression in MCF10A breast epithelial cells confers a premalignant phenotype that includes enhanced genomic instability and altered cell-cycle regulation. PMID- 17418865 TI - Reliability and usefulness of clinical encounter cards for a third-year surgical clerkship. AB - BACKGROUND: Third-year medical students (MS3) were given clinical encounter cards (CECs) to stimulate feedback during their surgery clerkship. This study analyzes the feedback given on their clinical performance using CECs. METHODS: Two hundred one students enrolled in the 12-week surgery clerkship were given CECs. Each card contained the chief focus of the encounter, which was rated on a six-point Likert scale. The CECs were analyzed to determine if they provided reliable formative information, identified marginal performances, and identified differences between raters, settings, rotations, and aspects of clinical performance evaluated. RESULTS: Seven thousand three hundred eight CECs were submitted from 201 students. The CECs were completed most often (65%) in the inpatient setting. Technical skills were evaluated on 49% of CECs, history/physical examinations on 40%, and case presentations on 30%. There were comments written on half of the CECs and 89% of these were strictly positive. Women (52.8%) and faculty (63.3%) were more likely to provide written comments. The students were most likely to seek feedback from the interns and faculty who gave significantly higher ratings. The Cronbach-alpha reliability coefficient was 0.69, based on 12 raters per student. There was a significant positive correlation (P < 0.05) between the CEC composite rating and the clinical performance rating (r = 0.356), NBME score (r = 0.258), and the final grade (r = 0.250). CONCLUSIONS: The implementation of CECs in a surgery clerkship provided a large quantity of positive feedback. The quality was limited because there were minimal recommendations for improvement and they were a poor predictor of overall clinical performance. PMID- 17418866 TI - Dual-color imaging of angiogenesis and its inhibition in bone and soft tissue sarcoma. AB - BACKGROUND: Angiogenesis is a critical step in tumor growth, progression, and metastasis. Soft tissue and bone sarcoma are resistant to most therapeutic approaches. Angiogenesis of these tumors may be an effective target. We hypothesized that we could inhibit tumor growth by targeting angiogenesis in a mouse model of sarcoma. We demonstrate in this report, using powerful color-coded fluorescent imageable tumor-host models, the onset of angiogenesis of these sarcomas and its inhibition. MATERIALS AND METHODS: Transgenic mice were used as the host in which green fluorescent protein (GFP) is driven by a regulatory element of the stem cell marker nestin (ND-GFP). Nascent blood vessels express ND GFP in this model. We visualized, by dual-color fluorescence imaging, angiogenesis of sarcoma formed by the HT-1080 human fibrosarcoma cell line expressing red fluorescent protein (RFP) in the ND-GFP mice. Tumor cells were injected into either the muscle or the bone. RESULTS: Nestin was highly expressed in proliferating endothelial cells and nascent blood vessels in the growing tumors, including the surrounding tissues. Immunohistochemical staining showed that CD31 colocalized in ND-GFP-expressing nascent blood vessels. The density of nascent blood vessels in the tumor was readily quantitated. The mice were given daily i.p. injections of 5 mg/kg of doxorubicin after implantation of tumor cells. Doxorubicin significantly decreased the mean nascent blood vessel density in the tumors as well as decreased tumor volume. CONCLUSION: The dual-color model of the ND-GFP nude mouse and RFP sarcoma cells is useful for the visualization and quantitation of bone and soft tissue tumor angiogenesis and evaluation of angiogenic inhibitors for such tumors. These data suggest targeting angiogenesis of sarcomas as a promising clinical approach. PMID- 17418867 TI - Loss of the tight junction protein ZO-1 in dextran sulfate sodium induced colitis. AB - BACKGROUND: Inflammatory bowel disease (IBD) is associated with increased intestinal permeability and decreased expression of tight junction (TJ) proteins in the inflamed mucosa. Whether this alteration in TJ expression is a prerequisite for the development of intestinal inflammation or a secondary result of that inflammation is unknown. This study looked at the expression of the TJ protein ZO-1 and the corresponding permeability changes in dextran sulfate sodium (DSS) induced colitis in a mouse model. MATERIALS AND METHODS: BALB/c mice were fed 3% DSS or water for 1, 3, 5, or 7 days. The animals were weighed, stool was checked for blood, and the colon length measured. Segments of the colon were used for histology, immunohistochemistry for ZO-1, or Western blot for TJ proteins. Colonic permeability was measured using Evan's Blue dye. RESULTS: DSS treated animals had heme positive stools, colitis by histology, significant weight loss, and colon shortening. There was an absence of ZO-1 by Western blot in the 7-day DSS treated animals, double the amount of claudin-1 and normal cytokeratin. The loss of ZO-1 started after 1 d of DSS treatment and was followed by a significant increase in permeability to Evan's blue by day 3. CONCLUSIONS: The loss of ZO-1 and increased permeability preceded the development of significant intestinal inflammation suggesting that in DSS colitis alterations in the TJ complex occur before the intestinal inflammation and not as a consequence of it. These changes in the TJ complex may facilitate the development of the inflammatory infiltrate seen in colitis. PMID- 17418868 TI - Adenoviral serotypes in gene therapy for esophageal carcinoma. AB - PURPOSE: Adenoviral gene therapy could potentially play a role in the treatment of esophageal cancer and Barrett's esophagus. The adenoviruses can be categorized in different serotypes. The goal of the present study was to investigate the transduction efficacy of different adenoviral serotypes in different models of esophageal cancer and Barrett's esophagus. METHODS: Chimeras of the adenoviral serotype 5 backbone and fibers of serotypes 5, 16, 35, 40, and 50 were constructed with PCR technology. For esophageal cancer, cell lines were used originating from with adenocarcinoma and squamous cell carcinoma, respectively. Differentiating Caco-2 cells were used as an in vitro model for Barrett's esophagus. GFP was used as a reporter gene and transduction efficacy was assessed by flow cytometry. RESULTS: Overall transduction was rather efficient in the cancer cell lines. Especially serotype 16 and 50 exhibited an improved transduction compared with the other serotypes. In the Caco-2 cell lines, we observed a decreased transduction upon differentiation of the cells. All serotypes had a very limited transduction and no serotype had an additional value in this setting. CONCLUSIONS: Some serotypes could have an additional value in the development of gene therapy for esophageal cancer. Especially serotype 16 and 50 exhibited an improved transduction in esophageal cancer cells compared with the native serotype 5. In the setting of Barrett's esophagus, none of the serotypes had an improved potency as in differentiated intestinal cells all serotypes had a very limited transduction. PMID- 17418869 TI - The use of adjusted preoperative CA 19-9 to predict the recurrence of resectable pancreatic cancer. AB - BACKGROUND: Despite the usefulness of CA 19-9 in the diagnosis and prognosis of pancreatic cancer, cholestasis can falsely elevate CA 19-9 levels, which contributes to limited clinical utility in patients with biliary obstruction. This study was designed to evaluate the usefulness of adjusted preoperative CA 19 9 levels in predicting a prognosis of pancreatic cancer. METHODS: The available medical records of patients with resected pancreatic cancer from January 1990 to June 2005 were retrospectively viewed at Yonsei Medical Center, Seoul, Korea. The adjusted CA 19-9 value was obtained by dividing the serum CA 19-9 level by the values of serum bilirubin in case of bilirubin greater, similar 2 mg/dL. Disease free survival was evaluated according to the adjusted preoperative CA 19-9 value. RESULTS: Sixty-one patients were investigated. Their adjusted preoperative CA 19 9 values were significantly different from the actual baseline CA 19-9 value (129.4 +/- 225.2 U/mL, versus 442.1 +/- 645.5 U/mL, P < 0.0001). On univariate analysis, peripancreatic microscopic invasion (P = 0.0142), lymphovascular invasion (P = 0.0038), and adjusted preoperative CA 19-9 > or = 50 U/mL (P = 0.0049) were predictive factors for cancer recurrence after curative resection. Adjusted preoperative CA 19-9 > or = 50 U/mL (Exp (B) = 2.097, P = 0.027) was an independent predictive factor in multivariate analysis. CONCLUSIONS: The adjusted preoperative CA 19-9 value can predict the risk of recurrence after curative resection of pancreatic cancer. Interpreting the preoperative CA 19-9 value adjusted to the serum bilirubin values seems to be more reasonable in evaluating prognosis of pancreatic cancer. PMID- 17418870 TI - Expression of vascular endothelial growth factor receptor-2 in the muscle flap with ischemic injury in rats. AB - BACKGROUND: Ischemia of muscle flaps is a significant problem in reconstructive surgery. Administration of either recombinant vascular endothelium growth factor (VEGF) protein or gene transfers has been conducted to improve the survival of ischemic tissue. These biophysiological functions of VEGF are mediated through its receptor. In this study the changes of expression in VEGF receptor mRNA in muscle flaps with acute ischemia were examined in a rat gracilis muscle flap model. MATERIALS AND METHODS: Twenty-four Sprague Dawley rats were used. The ischemia of the flap was induced by clamping its vascular pedicle. At 3-, 6-, and 18-h intervals, the flaps were biopsied for histology and examination of VEGFR-2 mRNA gene expression. The muscle flap without ischemia was used as the control. RESULTS: The results showed that the VEGFR-2 mRNA level was significantly increased 3 h after ischemia was induced and then attenuated to the baseline at 6 and 18 h ischemia. CONCLUSIONS: This study delineated the changes in expression of VEGFR-2 receptor in muscle flap in several postischemic intervals and provided a timeframe for the clinical application of VEGF. PMID- 17418871 TI - Nitric oxide donors selectively reduce the expression of matrix metalloproteinases-8 and -9 by human diabetic skin fibroblasts. AB - BACKGROUND: Chronic, nonhealing skin wounds are a common ailment in uncontrolled diabetes and are associated with significant morbidity. The nonhealing diabetic foot wound displays pathologically elevated matrix metalloproteinase (MMP) activity. In contrast, the concentration of nitric oxide (NO) is significantly reduced in these chronic ulcers. Addition of NO to diabetic wounds improves wound healing, but the mechanism for this effect is poorly understood. MATERIALS AND METHODS: Diabetic and nondiabetic human skin fibroblasts were cultured to confluence and then treated with 0, 1, 10, and 100 nm concentrations of three NO donors (NOR-3, SNAP, and SNOG) with varying half-lives for 1, 3, and 7 days. At harvest, the cultures were analyzed for their production of NO and the effect of NO donor treatment on cell proliferation (cell number) and MMP expression (MMP-1, -2, -8, -9, and -13). RESULTS: The NO donor with the shortest half-life (NOR-3) produced a rise in NO on day 1 in both normal and diabetic fibroblasts at the highest concentration used; there was a corresponding decrease in both MMP-8 and MMP-9 expression in the diabetic fibroblasts and a decrease in only MMP-9 expression in the normal fibroblasts. After longer times in culture or at lower concentrations, NOR-3 was without effect on NO production or MMP expression. Further, NOR-3 had no effect on cell proliferation. In contrast to NOR-3, NO donors with longer half-lives (SNAP and SNOG) significantly (P < 0.05) increased NO production by both normal and diabetic fibroblasts during the entire course of the experiment and even after a media change lacking additional NO donor at day 3. SNAP and SNOG dose-dependently reduced MMP-8 and -9 mRNA expression in both normal and diabetic fibroblasts through day 7. The expression of MMP-1, -2, and 13 was not significantly affected by any of the NO donor treatments. CONCLUSIONS: These experiments show distinct deficits in NO production and elevations in MMP-8 and -9 expression in diabetic human skin fibroblasts compared to normal. In addition, treating these cultures with NO donor compounds with half-lives greater than 5 h selectively raised NO production by the cells, increased cell proliferation, and decreased MMP-8 and -9 expression in a dose-dependent manner. There was no effect of the NO donor compounds on MMP-1, -2, or -13 expression. One possible mechanism to account for NO's beneficial effect on wound healing may involve stimulation of cell proliferation and down-regulation of MMP expression. PMID- 17418872 TI - Knot-tying force during suturing and wound healing in the gastrointestinal tract. AB - BACKGROUND: Objective data should be obtained to determine the most suitable knot tying force for appropriate suturing during robotic surgery. The purpose of this study was to estimate the appropriate force by investigating the efficiency of wound healing in the gastrointestinal tract in dog models. METHODS: We cut and sutured the stomach and jejunum of 12 beagles, using different magnitudes of knot tying forces. The relationship between the knot-tying force and variations in the density of the microvessels and basic-fibroblast growth factor (bFGF)-positive cells were evaluated. RESULTS: On the 7th postoperative day (POD) in the stomach, and the 7th and 11th POD in the jejunum, the density of the microvessels in the submucosa at the sites of cutting and ligation was higher for the knot-tying force of 1.5 N than for any other forces used. On the 4th and 7th POD in the stomach and 11th POD in the jejunum, the density of bFGF-positive cell in the mucosa was higher for the knot-tying force of 1.5 N than for any of the other forces used. CONCLUSIONS: By assessing the efficiency of wound healing, it was determined that the knot-tying force of 1.5 N may be the most appropriate for optimal wound healing in the gastrointestinal tract of the beagles. PMID- 17418873 TI - The role of quick bio-intact PTH(1-84) assay during parathyroidectomy for secondary hyperparathyroidism. AB - OBJECTIVE: The role of the quick PTH assay in surgery for secondary HPT is unclear because of overestimation of intact PTH(1-84) values due to the cross reactivity of currently available first-generation PTH assays with non-PTH(1-84) fragments that accumulate in renal failure. In this study, we used a second generation quick PTH immunometric assay that are claimed to detect the biologically active PTH(1-84) molecule with no cross-reactivity with PTH fragments to investigate the potential utility of the assay during parathyroidectomy for secondary HPT. MATERIAL AND METHODS: The study was performed on 18 patients (12 women, 6 men) between October 2004 and March 2005. EDTA serum samples were drawn via a peripheral venous catheter after induction of anesthesia (basal), and at 5, 10, and 30 min after excision of diseased parathyroid glands. Serum active PTH(1-84) was measured by the quick Bio-Intact PTH(1-84) assay, which is a two-site chemiluminometric assay. RESULTS: At 30 min the quick Bio-PTH(1-84) level of 16 patients was under 45 pg/mL. Four parathyroid glands were removed macroscopically from 12 of the 16 patients, and three glands were removed from the other four patients. All patients were cured of their HPT. Four enlarged parathyroid glands were removed from a patient whose Bio-Intact PTH(1-84) at 30 min had not fallen below 45 pg/mL, and no other glands were found by further exploration. At the 6 mo follow-up examination, the first-generation intact PTH level of this patient was over 45 pg/mL, but several diagnostic imaging methods did not reveal any enlarged parathyroid glands. Three enlarged parathyroid glands from the other patient, and exploration led to the identification of an ectopic parathyroid gland at the carotid bifurcation. CONCLUSIONS: The results of this prospective study show that quick Bio-Intact PTH(1-84) monitoring is a valuable new tool for use in the surgical treatment of secondary HPT. An intraoperative, quick Bio-Intact PTH(1-84) assay will be of value for the adequate prediction of surgical cure. PMID- 17418874 TI - The Estuarine Quality Paradox, Environmental Homeostasis and the difficulty of detecting anthropogenic stress in naturally stressed areas. AB - Estuaries have long been regarded as environmentally naturally stressed areas because of the high degree of variability in their physico-chemical characteristics, for example oxygen, temperature and salinity in the water column and bed sediment dynamics. However, their biota is well-adapted to cope with that stress and so the areas may be regarded as resilient because of that inherent variability; their ability to absorb stress without adverse effects is regarded here as Environmental Homeostasis. Hence these areas may only be regarded as stressful for marine or freshwater-adapted organisms and that for estuarine organisms this environmental stress is regarded as a subsidy whereby they successfully capitalise on the stressful conditions. In addition, using examples of the estuarine fauna and flora, this article indicates that the characteristics of natural stress in estuaries are similar to those for anthropogenic stress. An over-reliance on ecosystem structural features, such as diversity, in quality indicators therefore makes the detection of the anthropogenic stress more difficult. This difficulty is termed the Estuarine Quality Paradox. Because of these difficulties, the article argues that functional characteristics either as well as or rather than structural ones should be used in detecting environmental perturbations in estuaries. PMID- 17418875 TI - Decreased levels of lipid peroxidation-induced DNA damage in the onset of atherogenesis in apolipoprotein E deficient mice. AB - Increased oxidative stress and subsequent lipid peroxidation (LPO) are thought to be critical events in the formation of atherosclerotic lesions in apolipoprotein E deficient mice (ApoE-KO). LPO derived reactive aldehydes react with DNA to form exocyclic etheno-DNA adducts. These pro-mutagenic DNA lesions are known to be involved in the initiation of carcinogenesis, but their role in the development of atherosclerosis is unknown. In the present study we show that levels of the LPO derived 1,N(6)-ethenodeoxyadenosine (varepsilondA) and 3,N(4) ethenodeoxycytidine (varepsilondC) were both significantly lower in aorta of 12 weeks old ApoE-KO mice as compared to their wild type controls (1.6+/-0.3 versus 3.2+/-0.8 varepsilondA per 10(8) parent nucleotides, P=0.04 and 4.8+/-0.8 versus 9.2+/-2.1 for varepsilondC, P=0.02). Moreover, levels of both DNA adduct types were inversely related with total plasma cholesterol levels. Consequently, lowest etheno-DNA adduct levels were observed in ApoE-KO mice on a high fat diet. Hypercholesterolemia has previously been associated with increased expression of base excision repair (BER) enzymes, which could explain the lower levels of etheno-DNA adducts in ApoE-KO mice as compared to wild type controls. Indeed, increased staining for the BER-specific DNA repair enzyme apurinic/apyrimidinic endonuclease (Ape1/Ref1) was observed by immunohistochemistry in the endothelium and the first layers of arterial smooth muscle cells of ApoE-KO mice as compared to their wild type counterparts. A high fat diet further increased overall Ape1/Ref1 protein expression in ApoE-KO mice. Although these data suggest no role for increased LPO derived DNA damage in the onset of atherogenesis in ApoE-KO mice, the potentially modulating role of Ape1/Ref1 in the arterial wall deserves further attention. PMID- 17418876 TI - Inhibition of tyrosine kinase-mediated cellular signalling by Tyrphostins AG126 and AG556 modulates secondary damage in experimental spinal cord trauma. AB - Protein tyrosine kinases help to regulate the expression of many genes, which play an important role in the pathophysiology of a number of diseases. Here we investigate the effects of the tyrosine kinase inhibitors, AG126 and AG556 on the degree of experimental spinal cord trauma induced by the application of vascular clips to the dura via a four-level T4-T8 laminectomy. Spinal cord injury in mice resulted in severe trauma characterized by oedema, neutrophil infiltration, production of a range of inflammatory mediators, tissue damage, and apoptosis. Treatment of the mice with AG126 and AG556 significantly reduced the degree of (1) spinal cord inflammation and tissue injury (histological score), (2) neutrophil infiltration (myeloperoxidase activity), (3) iNOS, nitrotyrosine, and PARP expression and (4) apoptosis (TUNEL staining and Bax and Bcl-2 expression). In a separate set of experiments, AG126 and AG556 significantly ameliorated the recovery of limb function (evaluated by motor recovery score). This study provides an experimental evidence that (1) prevention of the activation of protein tyrosine kinases reduces the development of inflammation and tissue injury associated with spinal cord trauma, and (2) inhibition of the activity of certain tyrosine kinases may represent a novel approach for the therapy of spinal cord trauma. PMID- 17418877 TI - Short day-photoperiod triggers and low temperatures increase expression of peroxidase RNA transcripts and basic peroxidase isoenzyme activity in grapevine buds. AB - Expression of peroxidase (Prx) RNA transcripts was studied in grapevine cv. Thompson seedless through use of semi-quantitative RT-PCR. In roots, the three Vitis vinifera Prx ESTs, reported in the genomic facility database from the University of California (http://cgf.ucdavis.edu), were expressed as RNA transcripts, and three Prx isoenzymes, two basic (PrxB1) and (PrxB2) and a neutral Prx isoenzyme (PrxN1), were detected in root extracts. In buds, although one Prx EST (CB923206) was detected as a RNA transcript, three Prx activities were found in its extract, these same being the two basic isoenzymes found in the roots, and an acidic one (PrxA1) as well. In field grown grapevines, Prx RNA transcript was expressed transiently during bud endodormancy (ED) while under controlled conditions in a growth chamber, a short day (SD) photoperiod triggered expression of the Prx transcript in bud cuttings; low temperature, enhanced its expression level. Because environmental cues that trigger and enhance Prx RNA transcript expression in bud-tissue are the same as those that induce bud-ED in Vitis, the transient expression of Prx EST (CB923206) identified in buds could signal the full extent of ED in grapevines. PMID- 17418878 TI - The glucocorticoid receptor is increased in Atm-/- thymocytes and in Atm-/- thymic lymphoma cells, and its nuclear translocation counteracts c-myc expression. AB - We have previously demonstrated that spontaneous DNA synthesis in immature thymocytes of Atm-/- mice is elevated, and that treatment with the glucocorticoid dexamethasone (Dex) attenuates this increased DNA synthesis and prevents the development of thymic lymphomas. Deregulation of c-myc may drive the uncontrolled proliferation of Atm-/- thymocytes, since upregulation of c-myc parallels the elevated DNA synthesis in the cells. In this study, we show that the glucocorticoid receptor (GR) is expressed at high levels in Atm-/- thymocytes and in Atm-/- thymic lymphoma cells, although serum glucocorticoid (GC) levels in Atm /- mice are similar to those in Atm+/+ mice. In cultured Atm-/- thymic lymphoma cells treated with Dex, GR nuclear translocation occurs, resulting in suppression of DNA synthesis and c-myc expression at both the mRNA and protein levels. Interestingly, the GR antagonist RU486 also causes GR nuclear translocation, but does not affect DNA synthesis and c-myc expression in Atm-/- thymic lymphoma cells. As expected, RU486 reverses the suppressive effects of Dex on DNA synthesis and c-myc expression. Administration of Dex to Atm-/- mice decreases the elevated c-Myc protein levels in their thymocytes. These findings suggest that GC/GR signaling plays an important role in regulating c-myc expression in Atm-/- thymocytes and thymic lymphoma cells. PMID- 17418879 TI - Computational modelling of large aerated lagoon hydraulics. AB - A good understanding of the hydraulic performance of aerated lagoons is required for their design and operation. A comprehensive numerical procedure has been developed for the three-dimensional computational modelling of the flow in large lagoons including high-speed floating mechanical surface aerators. This paper describes the procedure that consists of separate aerator modelling, then applying the obtained results as boundary data for a full lagoon model. A model application to an industrial aerated lagoon serves as an example of flow analysis. Post processing of the results by calculating the local average residence time (age of fluid) provides a powerful and intuitive technique to visualize and analyse the lagoon performance. The model has been verified by comparing the local average residence time predictions with measurements from a dye study. It is shown that the numerical modelling proposed is feasible and constitutes an effective new tool in improving the performance and design of industrial lagoons. PMID- 17418880 TI - Sulfidogenic fluidized-bed treatment of metal-containing wastewater at 8 and 65 degrees C temperatures is limited by acetate oxidation. AB - Acetate utilization in sulfidogenic fluidized-bed reactors (FBRs) was investigated for the treatment of iron containing wastewater at low (8 degrees C) and high (65 degrees C) temperatures. The FBRs operated at low and high temperatures were inoculated with cultures of sulfate-reducing bacteria (SRB) originally enriched from arctic and hot mining environments, respectively. Acetate was not utilized as a carbon and electron source for SRB at 8 degrees C. With ethanol, hydrogen sulfide was produced from ethanol to acetate oxidation, which precipitated the iron. Then, several attempts were made to obtain acetate oxidation at 8 degrees C. Inoculation of two different low temperature enrichments and operating the FBR for a long period of time (321 days) did not result in enrichment of acetate oxidizing SRB. Due to the absence of acetate oxidation at 8 degrees C, external alkalinity addition was required to keep the pH neutral. At 65 degrees C, average acetate and sulfate removals were 52+/-12% and 24+/-8% at 670 mg/Ld acetate and 1500 mg/Ld sulfate loadings, respectively. The produced alkalinity from acetate oxidation increased the pH from 6.4 to around 7.5 and electron flow to sulfate reduction averaged 65%. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA genes showed quite stable SRB community at 8 degrees C, whereas, at 65 degrees C SRB community was dynamic. In the FBRs, Desulfomicrobium apsheronum and Desulfosporosinus sp. at 8 degrees C and Desulfotomaculum sp. at 65 degrees C were detected. PMID- 17418881 TI - Jack bean urease: the effect of active-site binding inhibitors on the reactivity of enzyme thiol groups. AB - In view of the complexity of the role of the active site flap cysteine in the urease catalysis, in this work we studied how the presence of typical active-site binding inhibitors of urease, phenylphosphorodiamidate (PPD), acetohydroxamic acid (AHA), boric acid and fluoride, affects the reactivity of enzyme thiol groups, the active site flap thiol in particular. For that the inhibitor-urease complexes were prepared with excess inhibitors and had their thiol groups titrated with DTNB. The effects observed were analyzed in terms of the structures of the inhibitor-urease complexes reported in the literature. We found that the effectiveness in preventing the active site cysteine from the modification by disulfides, varied among the inhibitors studied, even though they all bind to the active site. The variations were accounted for by different extents of geometrical distortion in the active site that the inhibitors introduced upon binding, leaving the flap either open in AHA-, boric acid- and fluoride-inhibited urease, like in the native enzyme or closed in PPD-inhibited urease. Among the inhibitors, only PPD was found to be able to thoroughly protect the flap cysteines from the further reaction with disulfides, this apparently resulting from the closed conformation of the flap. Accordingly, in practical terms PPD may be regarded as the most suitable inhibitor for active-site protection experiments in inhibition studies of urease. PMID- 17418882 TI - Effect of biosolid incorporation to mollisol soils on Cr, Cu, Ni, Pb, and Zn fractionation, and relationship with their bioavailability. AB - Biosolid application to soil may be a supply of nutrients and micronutrients but it may also accumulate toxic compounds which would be absorbed by crops and through them be incorporated to the trophic chain. The present study deals with the effect of biosolid application on Cr, Cu, Pb, Ni, and Zn in agricultural soils. The procedure used is sequential extraction so that the availability of those metals may be estimated and related to their bioavailability as determined through two indicator plants grown in greenhouse: ryegrass (Lolium perenne L.) and red clover (Trifolium pratense). Results showed that biosolid application to soil increased total Cu and Zn content. Sequential extraction showed that the more labile Zn fractions increased after biosolid application to soil. This was confirmed when assessing the total content of this metal in shoot and root of the plants under study, since a higher content was found in plant tissues, while no significant differences were found for Cu, Cr, Ni, and Pb. PMID- 17418883 TI - Modelling of the fate of selected endocrine disruptors in a municipal wastewater treatment plant in South East Queensland, Australia. AB - The aim of this study was to develop a fugacity-based analysis of the fate of selected industrial compounds (alkylphenols and phthalates) with endocrine disrupting properties in a conventional activated sludge wastewater treatment plant (WWTP A) in South East Queensland, Australia. Using mass balance principles, a fugacity model was developed for correlating and predicting the steady-state-phase concentrations, the process stream fluxes, and the fate of four phthalates and four alkylphenols in WWTP A. Input data are the compound's physicochemical properties, measured concentrations and the plant's operating design and parameters. The relative amounts of chemicals that are likely to be volatilized, sorbed to sludge, biotransformed, and discharge in the effluent water was determined. Since it was difficult to predict biotransformation, measured concentrations were used to calibrate the model in terms of biotransformation rate constant. Results obtained by applying the model for the eight compounds showed <40% differences between most of the estimated and measured data from WWTP A. All eight compounds that were modelled in this study had high removal efficacy from WWTP A. Apart from benzyl butyl phthalate and bisphenol A, the majority is removed via biotransformation followed by a lesser proportion removed with the primary sludge. Fugacity analysis provides useful insight into compound fate in a WWTP and with further calibration and validation the model should be useful for correlative and predictive purposes. PMID- 17418884 TI - The use of NTA and EDDS for enhanced phytoextraction of metals from a multiply contaminated soil by Brassica carinata. AB - The potential of nine different species to grow in the presence of metals (As, Cd, Cu, Pb and Zn) and to accumulate them in the shoots was assessed for each metal separately by germination and root length tests, and successively by hydroponic experiments. Of the nine species tested, Brassica carinata was the species that accumulated the highest amounts of metals in shoots without suffering a significant biomass reduction. To further evaluate the potential of B. carinata for chelant-enhanced phytoextraction of a natural, multiply metal polluted soil (As, Cd, Cu, Pb and Zn), both hydroponic and pot experiments were carried out with nitrilotriacetic acid (NTA) or (S,S)-ethylenediamine disuccinic acid (EDDS) as complexing agents. The hydroponic study with solutions containing the five metals together showed that accumulation of Cd, Cu, Pb and Zn in shoots was higher following EDDS addition compared to NTA. EDDS was more effective than NTA in desorbing Cu, Pb and Zn from the soil, whereas As and Cd were poorly extracted. B. carinata plants were grown for 4 weeks in the multiply metal contaminated soil and then the soil was amended with 5 mmol kg(-1) NTA or EDDS. All plants were harvested 1 week after amendment. In comparison to NTA, EDDS was more effective in enhancing the concentrations of Cu, Pb and Zn in B. carinata shoots (2- to 4-fold increase compared to the control). One week after chelant addition, the DTPA-extractable metal concentrations in the polluted soil were lower in the EDDS treatment in comparison with the NTA amendment. Even though B. carinata showed a reduced growth and a relatively low metal uptake, it demonstrated the ability to survive and tolerate the presence of more metals simultaneously. PMID- 17418885 TI - Inhibition of rainbow trout acetylcholinesterase by aqueous and suspended particle-associated organophosphorous insecticides. AB - Spraydrift and edge-of-field runoff are important routes of pesticide entry into streams. Pesticide contamination originating from spraydrift usually resides in the water phase, while pesticides in contaminated runoff are to a large extent associated with suspended particles (SPs). The effects of two organophosphorous insecticides (OPs), chloropyrifos (CPF) and azinphos-methyl (AZP), on acetylcholinesterase (AChE) activity in rainbow trout were compared between two exposure scenarios, simulating spraydrift- and runoff-borne contamination events in the Lourens River (LR), Western Cape, South Africa. NOECs of brain AChE inhibition, determined after 1h of exposure followed by 24h of recovery, were 0.33microgl(-1) for aqueous CPF, 200mgkg(-1) for SP-associated CPF and 20mgkg(-1) for SP-associated AZP (at 0.5gl(-1) SP). The highest aqueous AZP concentration tested (3.3microgl(-1)) was without significant effects. Previously reported peak levels of aqueous CPF in the LR ( approximately 0.2microgl(-1)) are close to its NOEC (this study), suggesting a significant toxicological risk to fish in the LR. By contrast, reported levels of SP-associated OPs in the LR are 20-200-fold lower than their NOECs (this study). In a comparative in situ study, trout were exposed for seven days at agricultural (LR2, LR3) and upstream reference (LR1) sites. No runoff occurred during the study. Brain AChE was significantly inhibited at LR3. However, OP levels at LR3 (CPF 0.01microgl(-1); AZP 0.14microgl(-1)) were minor compared to concentrations having effects in the laboratory (see above). Additionally, muscle AChE activity was significantly higher in caged trout from LR1 than in animals maintained in laboratory tanks. PMID- 17418886 TI - Consolidation of degraded ornamental porous limestone stone by calcium carbonate precipitation induced by the microbiota inhabiting the stone. AB - Although it has already been shown that calcareous stone can be consolidated by using a bacterially inoculated culture medium, a more user-friendly method is the in situ application of a sterile culture medium that is able to activate, among the microbial community of the stone, those bacteria with a potential for calcium carbonate precipitation. In order to test this new method for stone consolidation, non-sterilized decayed porous limestone was immersed in sterile nutritional media. Results were compared to those of the runs in which stone sterilized prior to the treatment was used. The effects of the microbial community on stone consolidation were determined by recording the evolution of the culture media chemistry. The treated stone was tested for mechanical resistance and porosity. Results demonstrate that the tested media were able to activate bacteria from the microbial community of the stone. As a consequence of the growth of these bacteria, an alkalinization occurred that resulted in calcium carbonate precipitation. The new precipitate was compatible with the substrate and consolidated the stone without pore plugging. Therefore, a good candidate to in situ consolidate decayed porous limestone is the application of a sterile culture medium with the characteristics specified in the present study. PMID- 17418887 TI - Hemispheric asymmetries in visual word-form processing: progress, conflict, and evaluating theories. AB - The ubiquitous left-hemisphere advantage in visual word processing can be accounted for in different ways. Competing theories have been tested recently using cAsE-aLtErNaTiNg words to investigate boundary conditions for the typical effect. We briefly summarize this research and examine the disagreements and commonalities across the theoretical perspectives. This work may provide a good example why a multi-level, multi-method, and multi-paradigm approach holds the greatest promise for rapid theoretical progress. PMID- 17418888 TI - Functional connectivity in an fMRI study of semantic and phonological processes and the effect of L-Dopa. AB - We describe an fMRI experiment examining the functional connectivity (FC) between regions of the brain associated with semantic and phonological processing. We wished to explore whether L-Dopa administration affects the interaction between language network components in semantic and phonological categorization tasks, as revealed by FC. We hypothesized that L-Dopa would decrease FC due to restriction of the semantic network. During two test sessions (placebo and L-Dopa) each participant performed two fMRI runs, involving phonological and semantic processing. A number of brain regions commonly activated by the two tasks were chosen as regions if interest: left inferior frontal, left posterior temporal and left fusiform gyri, and left parietal cortex. FC was calculated and further analyzed for effects of either the drug or task. No main effect for drug was found. A significant main effect for task was found, with a greater average correlation for the phonological task than for the semantic task. These findings suggest that language areas are activated in a more synchronous manner for phonological than for semantic tasks. This may relate to the fact that phonological processes are mediated to a greater extent within language areas, whereas semantic tasks likely require greater interaction outside of the language areas. Alternatively, this may be due to differences in the attentional requirements of the two tasks. PMID- 17418889 TI - Effects of short-term environmental disturbances on living benthic foraminifera during the Pacific oyster summer mortality in the Marennes-Oleron Bay (France). AB - Sediment cores were collected from April to August 2004 on tidal mudflats of the macrotidal Marennes-Oleron Bay (SW France), famous for the cultivation of Pacific oysters (Crassostrea gigas). The response of living (stained) benthic foraminifera to short-term biogeochemical disturbances in the sediment and overlying water, which may be involved in oyster summer mortality, was monitored. Short-term hypoxia occurred in early June, in conjunction with a sudden rise in temperature. In mid-June, the ammonia content of sediment porewater increased, leading to potentially maximal flux towards overlying waters. Foraminiferal assemblages, particularly in the topmost layer, were altered. Ammonia tepida was the most tolerant to temperature increase and hypoxic conditions whereas Brizalina variabilis and Haynesina germanica were sensitive to organic degradation and hypoxia. Cribroelphidium gunteri was the most opportunistic during recolonisation. Benthic foraminifera showed that short-term biochemical changes in the sediment are toxic and may be involved in the summer mortality of Pacific oysters. PMID- 17418890 TI - Low levels of copper reduce the reproductive success of a mobile invertebrate predator. AB - Marine organisms that occur in urbanised bays can be exposed to low-level chronic pollution that results in sublethal changes to behavior or reproduction. The effects of low levels of copper on the reproductive success of a mobile invertebrate were assessed. Free living flatworms are common predators of bivalves and barnacles. Flatworms (Stylochus pygmaeus) were exposed to low levels of copper ranging from 0 to 25 microg L(-1) in the presence and absence of their barnacle prey (Balanus variegatus). Flatworms laid fewer egg batches when exposed to copper and the hatching success of the eggs was also reduced. Exposure to 25 microg L(-1) copper for 10 d reduced the reproductive success of flatworms by up to 80%. Results were consistent regardless of the presence or absence of prey (barnacles). Barnacles were only moderately affected by copper but exhibited major avoidance behavior (feeding inhibition) in the presence of flatworm predators. This is the first ecotoxicological study on marine flatworms. Experiments are required to quantify the effects of flatworm predator populations on sessile invertebrate community structure in the field. PMID- 17418891 TI - SKY reveals a high frequency of unbalanced translocations involving chromosome 6 in t(12;21)-positive acute lymphoblastic leukemia. AB - The G-band cryptic t(12;21)(p13;q22) is the most common chromosomal rearrangement in childhood acute lymphoblastic leukemia (ALL). To investigate the nature of additional chromosomal events in this group of patients spectral karyotyping (SKY) following G-banding analysis was performed in 14 cases. From these cases six showed structural aberrations of chromosome 6, including both simple deletions and unbalanced translocations, and involved both q (n=4) and p (n=3) arms. The results show that rearrangements of 6p are also non-random events t(12;21)-positive ALL. This study illustrates the value of a combined SKY and G banding approach in identifying novel karyotypic events in childhood ALL. PMID- 17418892 TI - [Prevalence and characteristics of child sexual abuse among spanish university students]. PMID- 17418893 TI - Microcalorimetric study the toxic effect of hexavalent chromium on microbial activity of Wuhan brown sandy soil: an in vitro approach. AB - A multi-channel thermal activity monitor was applied to study soil microbial activity in Wuhan brown sandy soil in the presence of different concentrations of hexavalent chromium (K(2)Cr(2)O(7)). In order to stimulate the soil microbial activity, 5.0mg of glucose and 5.0mg of ammonium sulfate were added to a 1.20-g soil sample under a controlled humidity of 35%. The results show that the poisonous species of K(2)Cr(2)O(7) at an half inhibitory concentration (IC(50)) value of 4.27 microg mL(-1) against soil microbe, and an increase of the amount of hexavalent chromium is associated to a decrease in the microbial activity of the soil, probably due to an increase in the toxicity of hexavalent chromium, affecting strongly the life in this soil microbial environment. Our work also suggests that microcalorimetry is a fast, simple and more sensitive method that can be easily performed to study the toxicity of different species of heavy metals on microorganism compared to other biological methods. PMID- 17418894 TI - 5-HT(1A) receptors and memory. AB - The study of 5-hydroxytryptamine (5-HT) systems has benefited from the identification, classification and cloning of multiple 5-HT receptors (5-HT(1)-5 HT(7)). Increasing evidence suggests that 5-HT pathways, reuptake site/transporter complex and 5-HT receptors represent a strategic distribution for learning and memory. A key question still remaining is whether 5-HT markers (e.g., receptors) are directly or indirectly contributing to the physiological and pharmacological basis of memory and its pathogenesis or, rather, if they represent protective or adaptable mechanisms (at least in initial stages). In the current paper, the major aim is to revise recent advances regarding mammalian 5 HT(1A) receptors in light of their physiological, pathophysiological and therapeutic implications in memory. An attempt is made to identify and discuss sources of discrepancies by employing an analytic approach to examine the nature and degree of difficulty of behavioral tasks used, as well as implicating other factors (for example, brain areas, training time or duration, and drug administration) which might offer new insights into the understanding and interpretation of these data. In this context, 8-OH-DPAT deserves special attention since for many years it has been the more selective 5-HT drug and, hence, more frequently used. As 5-HT(1A) receptors are key components of serotonergic signaling, investigation of their memory mechanisms and action sites and the conditions under which they might operate, could yield valuable insights. Moreover, selective drugs with agonists, neutral antagonists or inverse agonist properties for 5-HT(1A) (and 5-HT(7)) receptors may constitute a new therapeutic opportunity for learning and memory disorders. PMID- 17418895 TI - Highly diverse TCR delta chain repertoire in bovine tissues due to the use of up to four D segments per delta chain. AB - Tissue-specific distribution of gammadelta TCRs with limited TCR diversity is a common phenomenon in species with a low percentage of gammadelta T cells like humans and mice. We set out to investigate whether this is also the case in cattle (Bos taurus), a species with high percentages of gammadelta T cells. Using a method that was independent of variable (V) segment-specific primers, we generated 65 unique TCR delta chain sequences. We found no evidence for preferential use of certain Vdelta segments in lymph node, skin, spleen, small intestine, large intestine, and blood. The delta chain CDR3 length distribution was very wide in each tissue, which was confirmed by spectratyping. The highly variable CDR3 length was due to the use of up to four diversity (D) segments by one bovine delta chain. Human and murine delta chains contain only one or two D segments. The five functional Ddelta segments that we describe here were identified at cDNA and genomic level, and are the first ruminant D segments described. Fourteen TCR delta chain sequences used novel Vdelta1 segments, and one expressed a novel member of the Vdelta3 family. The number of known functional Vdelta segments in cattle including these new ones is 42 now, but the total number may be much higher. A high number of Vdelta segments in combination with the use of up to four out of five D segments, and the possibility of using non-template encoded (N) nucleotides on either side of these, makes the potential bovine delta chain repertoire much bigger than any known TCR chain. This situation is quite different from the situation in humans and mice, and suggests that the differences between gammadelta high and gammadelta low species in distribution, diversity, and function of gammadelta T cells may be substantial. PMID- 17418896 TI - Beta-arrestins 1 and 2 differentially regulate LPS-induced signaling and pro inflammatory gene expression. AB - Toll like receptors, the critical receptor family in innate immunity, have been shown to signal via both ERK 1/2 and transcription factor NFkappaB. beta Arrestins 1 and 2 have recently been implicated in modulation of NFkappaB signaling and ERK 1/2 activation. Using a number of approaches: mouse embryonic fibroblasts (MEF) from wild-type (WT), beta-arrestins knockouts (KO), beta arrestins 1 and 2 double KO, and MEFs with reconstituted WT beta-arrestins in the double KO cells, RNA interference (siRNA) specific knockdown of beta-arrestins, and overexpression of WT beta-arrestins, it was demonstrated that beta-arrestin 2 positively regulates LPS-induced ERK 1/2 activation and both beta-arrestins 1 and 2 negatively regulate LPS-induced NFkappaB activation. Also beta-arrestin 2 positively regulate LPS-induced IL-6 production and both beta-arrestins 1 and 2 positively regulate LPS-induced IL-8 production. The specific ERK1/2 inhibitor PD98059 significantly decreased LPS-induced IL-6 and IL-8 production suggesting that IL-6 and IL-8 production is, in part, mediated by ERK 1/2 activation. Over expression of wild type beta-arrestins 1 and 2 had no effect on LPS-induced ERK1/2 activation and LPS-induced IL-8 production suggesting that endogenous beta arrestins 1 and 2 are sufficient to mediate maximum ERK 1/2 activity and IL-8 production. beta-Arrestins thus not only negatively regulate LPS-induced NFkappaB activation but also positively regulate ERK 1/2 activation and specific pro inflammatory gene expression. Understanding the role of beta-arrestins in regulation of TLR signaling pathways may provide novel insights into control mechanisms for inflammatory gene expression. PMID- 17418897 TI - Identification and characterization of a cDNA encoding a crustin-like, putative antibacterial protein from the American lobster Homarus americanus. AB - Pathogenic challenges in decapod crustaceans are combated by innate immune responses, including the production and secretion of soluble antibacterial proteins into the hemolymph. Among the antibacterials that have been identified in decapod species are the crustins, a group of four-disulfide core/whey-acidic protein (WAP) domain-containing proteins, which target marine/salt tolerant Gram positive bacteria. To begin to assess the possible role of crustins in combating bacterial invasion in the American lobster Homarus americanus, we identified and sequenced a 744 base pair cDNA that encodes a novel 96 amino acid crustin-like protein. Comparison of H. americanus crustin (Hoa-crustin) with crustins from other decapod species showed that it is most similar to an isoform predicted from the European lobster Homarus gammarus ( approximately 86% identity). With our identification of the Hoa-crustin cDNA, we are positioned to begin molecular and physiological investigations of the regulation and function of this putative antibacterial protein in H. americanus. PMID- 17418898 TI - Plasma activity of thrombin activatable fibrinolysis inhibitor in Crimean-Congo hemorrhagic fever. AB - OBJECTIVES: Crimean-Congo hemorrhagic fever (CCHF) is a potentially fatal viral infection. The exact mechanism for hemorrhage remains unknown. Thrombin activatable fibrinolysis inhibitor (TAFI) is a plasma procarboxypeptidase B-like proenzyme and synthesized in the liver, down-regulating fibrinolysis. In this study, we measured the TAFI activity in plasma of patients with CCHF to examine the relationship between hemorrhage and the pathogenesis of CCHF. METHODS: Twenty one patients and similar number of controls were included in the study. The diagnosis of CCHF was confirmed through detection of IgM by ELISA and/or PCR. TAFI activity was measured in plasma samples. RESULTS: TAFI activity in CCHF patient group was mean 7.2+/-2.3 microg/ml (range: 0.95-10.31 microg/ml) and in the control group was mean 11.7+/-4.1 microg/ml (range: 3.07-23.9 microg/ml). There was a significant decrease of TAFI activity in CCHF patients when compared to controls. A positive correlation between CRP, PT, INR, serum albumin and TAFI activity levels were found. We suggest that the decrease of TAFI activity may be due to liver dysfunction during viral active disease state. CONCLUSIONS: Low TAFI activity may be an attributable factor, leading to imbalance in fibrinolysis, resulting in bleeding complications in CCHF. PMID- 17418899 TI - A simple indicator of nonstationarity of firing rate in spike trains. AB - The assessment of stationarity of firing rate in neural spike trains is important but is often performed only visually. Facing the growing amount of neural data generated by multi-electrode recording, there is a need for an automatic method to identify and disqualify spike trains with highly nonstationary firing rates. In this report, we propose a simple test of nonstationarity, associated with an indicator quantifying the degree of nonstationary in a spike train. This method is compared to the Mann-Kendall test of trend detection and the Runs test on simulated and real spike trains. PMID- 17418900 TI - Anterograde tracing method using DiI to label vagal innervation of the embryonic and early postnatal mouse gastrointestinal tract. AB - The mouse is an extremely valuable model for studying vagal development in relation to strain differences, genetic variation, gene manipulations or pharmacological manipulations. Therefore, a method using 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) was developed for labeling vagal innervation of the gastrointestinal (GI) tract in embryonic and postnatal mice. DiI labeling was adapted and optimized for this purpose by varying several facets of the method. For example, insertion and crushing of DiI crystals into the nerve led to faster DiI diffusion along vagal axons and diffusion over longer distances as compared with piercing the nerve with a micropipette tip coated with dried DiI oil. Moreover, inclusion of EDTA in the fixative reduced leakage of DiI out of nerve fibers that occurred with long incubations. Also, mounting labeled tissue in PBS was superior to glycerol with n propyl gallate, which resulted in reduced clarity of DiI labeling that may have been due to DiI leaking out of fibers. Optical sectioning of flattened wholemounts permitted examination of individual tissue layers of the GI tract wall. This procedure aided identification of nerve ending types because in most instances each type innervates a different tissue layer. Between embryonic day 12.5 and postnatal day 8, growth of axons into the GI tract, formation and patterning of fiber bundles in the myenteric plexus and early formation of putative afferent and efferent nerve terminals were observed. Thus, the DiI tracing method developed here has opened up a window for investigation during an important phase of vagal development. PMID- 17418901 TI - Comparison of two methods for quantitative assessment of unrestrained locomotion in the rat. AB - Changes in locomotor movements induced by central and peripheral nerve injury or obtained as a result of pharmacological treatment are increasingly being investigated in rats. Several methods have been used to assess changes in the main locomotor indices, most of which are based on video recordings, usually with low time resolution, or on X-ray cinematographic recordings. Other methods are based on qualitative visual locomotor scoring systems like the BBB scale. We have analyzed locomotor indices in freely moving rats using two methods that can give quantitative results and which may be readily automated. One is based on detecting the onsets of swing and stance phases with contact electrodes (CE), while the second is based on recording the bursts of electromyographic activity (EMG) from the flexor and extensor muscles of each limb during the swing and stance phases, respectively. Besides the investigation of spontaneous locomotion in intact rats, our study also included an examination of locomotion on a ladder using EMG recording and analysis of locomotor disturbances following spinal cord hemisection, for which combined application of the two methods appeared to be useful. Overall, the EMG method appears to be more versatile than the CE method, although the use of both methods in parallel is recommended. PMID- 17418902 TI - Biochemical property and immunogenicity of mouse male reproductive tract CD52 (mrt-CD52). AB - Male reproductive tract CD52 (mrt-CD52) is known to be a pathogenic antigen for immunological infertility. Although human CD52 has been extensively investigated, the properties of mouse CD52 are not well elucidated. This study was conducted, therefore, to examine the tissue distribution, molecular composition and immunogenicity of mouse mrt-CD52. Immunohistological studies with an antibody to a synthetic peptide showed that mouse CD52 was localized mainly in the cauda epididymis and vas deferens, but not in the testis, liver, kidney or spleen. The molecule was composed of Asn (N)-linked and The/Ser (O)-linked carbohydrates as well as a glycosylphosphatidyl (GPI) anchor portion. Purified mrt-CD52 preparations produced antibodies by subcutaneous and intranasal immunization in both male and female mice. These antisera showed sperm-immobilizing activities with complement to mouse sperm. The research indicated mouse CD52 had similar biochemical and immunological properties to human CD52. This animal experiment is a good model for investigating human mrt-CD52 antibody detected in infertile patients. PMID- 17418903 TI - ZP3 peptides administered orally suppress murine experimental autoimmune ovarian disease. AB - Experimental autoimmune ovarian disease (AOD) is a T cell-mediated chronic inflammatory disease that may lead to premature ovarian failure. Autoimmune disease can be suppressed by oral administration of autoantigens leading to tolerance. One of the major mechanisms of oral tolerance is induction of regulatory CD4+ T cells that can mediate active suppression by producing immunomodulatory cytokines. However, the role of oral tolerance as a treatment for experimental AOD has received little attention. Therefore, the purpose of this study was to examine the conditions necessary to produce oral tolerance in experimental AOD in B6AF1 female mice. In this study, mice received different doses of peptides of the mouse zona pellucida 3 (pZP3) via gastric intubation for 7 times. After 4 times of oral administration, AOD was induced by immunization with pZP3. The optimal tolerating regimen for oral administration of pZP3 in mice was 10 microg, which decreased morbidity of oophoritis compared to the control group. In this moderate-dose therapeutic group (MD), alterations in the estrous cycle were normalized and CD4+ T cells that were CD25+ increased while those that were CD25- decreased. The severity of autoimmune oophoritis and the titer of ZP autoantibodies were also significantly reduced. These findings suggest that oral administration of pZP3 may be successfully used as an oral tolerance strategy for suppression of AOD. PMID- 17418904 TI - NMDA and AMPA receptors: old channels, new tricks. AB - Learning and memory depend on persistent changes in synaptic strength that require neuronal gene expression. An unresolved question concerns the mechanisms by which activity at synapses is transduced into a nuclear transcriptional response. In the prevailing view, N-methyl-D-aspartate (NMDA)- and alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-type glutamate receptors have distinct roles in controlling synaptic strength: AMPA receptors effect short-term changes in synaptic strength, whereas NMDA receptors regulate genes that are required for the long-term maintenance of these changes. Here, we review recent data on the roles of these two types of receptor in activity-dependent gene expression. We discuss evidence that signals from NMDA receptors and AMPA receptors are integrated to specify transcriptional responses for particular plasticity related genes. PMID- 17418905 TI - The topological role of homeoproteins in the developing central nervous system. AB - Homeogenes encode homeoprotein transcription factors that have fundamental roles in development. They are key players in genetic networks that lay out the body plan and also determine morphology and physiology at the cellular and multicellular level. However, homeoproteins share activities that extend beyond transcription, including translation regulation and signalling. For example, homeoproteins participate in the definition of territories in the neuroepithelium and also have a function in axonal guidance. Based on these examples, we propose that homeoproteins are not only morphogenetic transcription factors, but also morphogens themselves. PMID- 17418906 TI - Extensive heterozygosity at four microsatellite loci flanking Plasmodium vivax dihydrofolate reductase gene. AB - Only limited but contrasting reports are available on microsatellites based population structure of Plasmodium vivax. Further, there is complete lack of information on microsatellites in the flanking regions of the P. vivax drug resistance genes to trace the origin and spread of the drug resistant vivax malaria. Therefore, we scanned +/-300 kb flanking sequences of the P. vivax dihydrofolate reductase (pvdhfr) gene for di-nucleotide microsatellite repeats with minimum of 8 unit array length. Only 13 such repeats were detected in this region as compared to 738 di-nucleotide repeats present in +/-300 kb flanking region of P. falciparumdhfr gene. We have analyzed here the nucleotide sequence of 110 Indian P. vivax isolates for four of these di-nucleotide microsatellites (two in the nearest regions at -38.83 kb and +6.15 kb, and two in the farthest regions at -230.54 kb and +283.28 kb). All the four microsatellites were found to be highly polymorphic in the population where number of alleles varied from 4 to 10 with the median values of 9-11 at these loci. The expected heterozygosity (He) at these loci ranged from 0.50 to 0.82. We did not find any association between pvdhfr mutations and the flanking microsatellite alleles. There was a regional variation in the microsatellites polymorphism which was not associated with the reported prevalent rates of drug resistance or malaria transmission. In conclusion, the level of microsatellite polymorphism in P. vivax is as high as in P. falciparum. These results will be valuable in understanding the evolutionary history of the pvdhfr alleles as well as for designing the malaria control strategies. PMID- 17418907 TI - Increased efficiency of homologous recombination in Toxoplasma gondii dense granule protein 3 demonstrates that GRA3 is not necessary in cell culture but does contribute to virulence. AB - Toxoplasma gondii possesses unique secretory organelles, which synchronously release proteins during and after invasion. One of these organelles, the dense granules, secrete proteins after invasion which are thought to be important in development of the parasite throughout all stages of its life cycle. Dense granule protein 3 (GRA3) is a 30 kDa protein localized to the intravacuolar network and parasitophorous vacuole membrane (PVM). Like many dense granule proteins, GRA3 has no homology to proteins with described functions. However, it has been hypothesized to be involved in nutrient acquisition for the parasite due to its localization on the PVM. To begin to investigate the importance of GRA3, the locus was disrupted by homologous replacement with a chloramphenicol resistance gene in a type II strain. Two DeltaGRA3 strains were obtained after two independent electroporations with efficiency greater than 80%. No differences between wild-type and DeltaGRA3 were detected in cell culture growth rate or bradyzoite formation. Location of other parasite dense granule proteins and association with host cell organelles were also not affected in DeltaGRA3. Interestingly, at an infectious dose approximately four-fold above the lethal dose 50% for wild-type parasites, all mice infected with DeltaGRA3-2 infected mice survived acute infection. Complementation of GRA3 expression in the DeltaGRA3-2 strain restored virulence to wild-type levels, and increased the virulence of the DeltaGRA3-1, confirming that the GRA3 protein plays a role during acute infection in a type II strain. PMID- 17418908 TI - Catechin combined with vitamins C and E ameliorates insulin resistance (IR) and atherosclerotic changes in aged rats with chronic renal failure (CRF). AB - Aging is an inevitable biological process associated with increased oxidative stress and accumulation of asymmetric dimethylarginine (ADMA) a known endogenous inhibitor of nitric oxide synthase. Atherosclerosis and IR constitute major risk factors for cardiovascular mortality in elderly with chronic kidney disease (CKD). We investigated the impact of catechin, vitamins E and C supplementation on insulin sensitivity, redox state, ADMA, nitrate and nitrite (NO(2)(-)/NO(3)( )) levels and histological picture of heart and large blood vessels of aged rats with CRF. Findings of the present study revealed that aging in rats is associated with hyperinsulinemia, hyperlipidemia, IR indicated by higher homeostasis model assessment (HOMA)-index, increased lipid peroxidation product malondialdehyde (MDA), ADMA, and blood pressure (BP), but decreased antioxidant capacity and NO(2)(-)/NO(3)(-) levels. CRF exaggerated all these findings and caused thickened intima of carotid arteries and myocardial hypertrophy. Treatment with catechin, vitamins E and C increases the antioxidant capacity and NO(2)(-)/NO(3)(-) production but, decreases MDA, ADMA and BP levels. Also it keeps insulin sensitivity and normal intima/media thickness of carotid arteries. We conclude that decreased nitric oxide (NO) availability due to ADMA accumulation may be responsible for IR and associated atherosclerotic changes in aged rats with CRF. Catechin, vitamins E and C supplementation may moderate oxidative stress of renal failure, prevent ADMA accumulation, and counteract IR and atherosclerotic changes in the elderly. PMID- 17418909 TI - PC12 cell model of inducible expression of mutant DISC1: new evidence for a dominant-negative mechanism of abnormal neuronal differentiation. AB - A balanced chromosomal translocation, segregating with mental illnesses in a large Scottish family, interrupts the disrupted-in-schizophrenia 1 (DISC1) gene, which would result in loss of DISC1 function via haploinsufficiency or dominant negative effects (or possibly could cause gain-of-function effects) if a truncated protein is present. To evaluate the effects of a predicted protein, mutant DISC1, we generated stable PC12 cell clones with inducible expression of mutant or full-length human DISC1 (hDISC1). Our study presents new observations that the inhibitory effects of mutant hDISC1 on NGF-induced neurite outgrowth are dependent on the level and timing of expression of mutant DISC1 and the concentrations of NGF, and are associated with altered sub-cellular distribution of endogenous DISC1 and ATF4, and decreased protein levels of LIS1. Thus, inducible expression of DISC1 in PC12 cell clones is a valuable in vitro model for further studying the molecular mechanisms likely due to loss of function of DISC1 relevant to the pathogenesis of major mental illnesses. PMID- 17418910 TI - Fibrillogenic amylin evokes the apoptosis of human mesangial cells. AB - OBJECTIVE: To explore the apoptotic role of amylin on human mesangial cell (MC). MATERIALS AND METHODS: Primarily cultured human MCs were applied and treated with fresh amylin preparation. Human MCs were identified by the morphology and immunofluorescence staining. The apoptotic cells were determined by ultrastructure changes, TUNEL, and DNA fragmentation analysis. Propidium iodide staining and flow cytometry was employed for quantitative measurement of apoptosis. RESULTS: Under the light and transmission electronic microscopy (TEM), the human MCs with condensed chromatin, plasma shrinkage, marginated nuclear chromatin or apoptotic body were observed in amylin-treated MCs. Positive TUNEL staining, hypolipoid DNA peak, and typical DNA "ladder" pattern were also detected in amylin-treated MCs. Quantitative analysis of the apoptotic MCs showed that human amylin induced an increase of the percentage of apoptotic cells in a dose-dependent manner. Amylin nano-scale fibrils (5-18nm) in diameter were detected in the cultured solution using negative staining under the TEM. Compared to the control, no significant changes of lactate dehydrogenase release were observed in amylin-treated MCs (P>0.05). CONCLUSIONS: Fibrillogenic amylin evokes the apoptosis of human MCs in vitro, which may explain the mechanism of the hypocellular mesangial damage and progressive glomerulosclerosis of the patients with diabetic nephropathy. PMID- 17418911 TI - The pattern and evolution of yeast promoter bendability. AB - In Saccharomyces cerevisiae, transcription factor binding sites are found preferentially approximately 100-200 bp upstream of the start codon. Here, we show that this region is associated with rigid DNA in promoters lacking a TATA box, but not in TATA-containing promoters. The association of rigid DNA with transcription factor binding sites is conserved in TATA-less promoters from 11 yeast species, whereas the position of the rigid DNA varies substantially among species. Rigid DNA could influence nucleosome positioning and assist in the assembly of the transcriptional machinery at TATA-less promoters. PMID- 17418912 TI - Distinct claudin expression profile in histologic subtypes of lung cancer. AB - PURPOSE: Claudins are the main constituents of tight junctions. Little is known about their expression and localization in the normal bronchial epithelium and in lung cancer. PATIENTS AND METHODS: One hundred four lung cancer tissue blocks were studied including 46 adenocarcinomas (ADC), 30 squamous cell carcinomas (SCC), 15 small cell lung cancers (SCLC), 8 typical and 5 atypical carcinoids. All slides contained normal bronchial mucosa as well. Immunohistochemistry using antibodies against claudins-1, -2, -3, -4, and -7 proteins, as well as semi quantitative estimation were performed. RT-PCR analysis was also carried out in 22 immunohistochemically representative tumor samples. RESULTS: Normal bronchial epithelial cells expressed all the examined claudin proteins. When compared, SCLCs and carcinoids showed striking differences in regard to claudins-1, -3, and -4 expressions (p<0.0001, p<0.0001, and p<0.0004, respectively), whereas ADCs and SCCs revealed significant differences in claudins-3, -4, and -7 expressions (p<0.0001, p<0.0001, and p<0.0053, respectively). However, comparison of ADCs with SCLCs revealed significant difference only in claudin-2 expression (p<0.0002). The comparison of ADCs and carcinoids resulted in significant differences regarding claudins-1, -3, and -4 expressions (p<0.0006, p<0.0001, and p<0.0001, respectively). SCCs and SCLCs varied in respect to claudin-2, -3, and 4 expressions (p<0.0009, p<0.0001, and p<0.0019, respectively), whereas SCCs and carcinoids showed different claudins-1 and -4 expressions (p<0.0076 and p<0.0045, respectively). RT-PCR analysis revealed parallel changes in the mRNA and protein expression of certain claudins. CONCLUSIONS: The observed distinct claudin expression profile within the non-small cell lung cancer group, further, the marked differences between SCLCs and carcinoids may have differential diagnostic impact, and the overexpression of certain claudins might have therapeutic implications. PMID- 17418913 TI - Role of plant lipid transfer proteins in plant cell physiology-a concise review. AB - Plant lipid transfer proteins (LTP) are cationic peptides, subdivided into two families, which present molecular masses of around 7 and 10 kDa. The peptides were, thus, denominated due to their ability to reversibly bind and transport hydrophobic molecules in vitro. Both subfamilies possess conserved patterns of eight cysteine residues and the three-dimensional structure reveals an internal hydrophobic cavity that comprises the lipid binding site. Based on the growing knowledge regarding structure, gene expression and regulation and in vitro activity, LTPs are likely to play a role in key processes of plant physiology. Although the roles of plant LTPs have not yet been fully determined. This review aims to present comprehensive information of recent topics, cover new additional data, and present new perspectives on these families of peptides. PMID- 17418914 TI - Association of a NOS1 promoter repeat with Alzheimer's disease. AB - The gene encoding NOS-I (NOS1) displays a complex transcriptional regulation, with nine alternative first exons. Exon 1c and 1f are the most abundant forms in the brain. A functional single nucleotide polymorphism (SNP) in exon 1c and a polymorphism in exon 1f, consisting of a variable number of tandem repeats (VNTR) originating short (S) and long (L) alleles, were studied in 184 patients with Alzheimer's disease (AD) and 144 gender- and age-matched controls. No differences were found for the Ex1c G-84A. The Ex1f-VNTR S allele was significantly more common in AD (55% versus 44%, P=0.009, OR=1.52) as was the S/S genotype (28% versus 14%, P=0.008; OR=2.37). The S allele showed a highly significant interaction with the ApoE epsilon 4 allele (OR: 10.83). Therefore, short alleles of the NOS1 exon 1f-VNTR are likely to be susceptibility factors for AD, and interact with the epsilon 4 allele to markedly increase the AD risk. PMID- 17418915 TI - QSAR study on a novel series of 8-azaadenine analogues proposed as A1 adenosine receptor antagonists. AB - 8-Azaadenines have been recently proposed as a novel promising class of adenosine A1 receptor antagonists. A QSAR study on 45 derivatives, synthesized in our laboratory as antagonists for A1 receptor, is described here. The use of the CODESSA program allowed obtaining a quite simple equation capable of correlating the structural features of these ligands to their activity toward A1 receptor. The model was investigated for reliability and stability by using statistical analysis criteria stricter than usual. Particular care was put in defining the chemical space where the model gave reliable predictions. The model allowed the identification of relevant structural features required for the interaction with the A1 receptor, enabling the prediction of activity of molecules belonging to focused virtual libraries. PMID- 17418916 TI - Synthesis and antileishmanial activity of 6-mono-substituted and 3,6-di substituted acridines obtained by acylation of proflavine. AB - Two new series of diaminoacridinic derivatives obtained from proflavine and N-(6 amino-3-acridinyl)acetamide were synthesised and assessed for their cytotoxic and antileishmanial activities. Two compounds, N-[6-(acetylamino)-3 acridinyl]acetamide and N-[6-(benzoylamino)-3-acridinyl]benzamide demonstrated highly specific antileishmanial properties against the intracellular amastigote form of the parasite. Structure-activity relationships established that the antiproliferative activity against human cells was greatly enhanced by the presence of a benzoylamino group in 6-mono-substituted acridines, while the presence of two acetylamino or benzoylamino groups in 3,6-di-substituted acridines strongly increased the specificity of the molecules for Leishmania parasite, suggesting that symmetric conformations could preferentially interfere with Leishmania metabolism. PMID- 17418917 TI - Poor agreement between continuous measurements of energy expenditure and routinely used prediction equations in intensive care unit patients. AB - BACKGROUND & AIMS: A wide variation in 24h energy expenditure has been demonstrated previously in intensive care unit (ICU) patients. The accuracy of equations used to predict energy expenditure in critically ill patients is frequently compared with single or short-duration indirect calorimetry measurements, which may not represent the total energy expenditure (TEE) of these patients. To take into account this variability in energy expenditure, estimates have been compared with continuous indirect calorimetry measurements. METHODS: Continuous (24h/day for 5 days) indirect calorimetry measurements were made in patients requiring mechanical ventilation for 5 days. The Harris-Benedict, Schofield and Ireton-Jones equations and the American College of Chest Physicians recommendation of 25 kcal/kg/day were used to estimate energy requirements. RESULTS: A total of 192 days of measurements, in 27 patients, were available for comparison with the different equations. Agreement between the equations and measured values was poor. The Harris-Benedict, Schofield and ACCP equations provided more estimates (66%, 66% and 65%, respectively) within 80% and 110% of TEE values. However, each of these equations would have resulted in clinically significant underfeeding (<80% of TEE) in 16%, 15% and 22% of patients, respectively, and overfeeding (>110% of TEE) in 18%, 19% and 13% of patients, respectively. CONCLUSIONS: Limits of agreement between the different equations and TEE values were unacceptably wide. Prediction equations may result in significant under or overfeeding in the clinical setting. PMID- 17418918 TI - Combined virus-like particle-based polyepitope DNA/protein HIV-1 vaccine design, immunogenicity and toxicity studies. AB - We have previously described designing of polyepitope immunogens TBI and TCI, to stimulate the humoral and cellular immune responses to HIV-1. Here, immunogens TBI and TCI were used to create new vaccine construct named CombiHIVvac (Combined HIV-1 vaccine). CombiHIVvac is a virus-like particles (VLP) containing the DNA vaccine pcDNA-TCI as a core encapsulated within a spermidine-polyglucin-TBI conjugate. The immunogenic and toxic properties of the candidate vaccine CombiHIVvac have been studied. CombiHIVvac induces a strong humoral and CTL responses in mice; the antibodies are highly specific and are able to neutralize HIV-1 in vitro. Preclinical study demonstrated that CombiHIVvac does not cause long-term changes in physiological, biochemical and morphological parameters in immunized animals and thus can be recommended for clinical trials. PMID- 17418919 TI - Modeling 137Cs migration processes in lake sediments. AB - Lake sediments can be considered as the "diary" of the lake: new layers are continuously deposited on top of the sediments. The migration behavior of radionuclides, particularly of (137)Cs, can be analyzed by modeling the input into and the vertical distribution within the sediment. For this purpose, a model consisting of sedimentation-diffusion equations was developed. Solved with a finite element method, this model can cover the time period from the nuclear weapons testing to the present. It takes into account fixation and redissolution, compaction of sediments, and the influence of competing ions on the retarded diffusion within the sediments. The outcome of the model is compared to water and sediment measurements from Lago Maggiore. The results of an optimization process can produce the sedimentation rate and (137)Cs distribution coefficients which determine the uptake of activity into the sediment and also the retarded diffusion within the sediment. PMID- 17418920 TI - Fate of HTO following its acute soil deposition at different growth stages of Chinese cabbage. AB - Lysimeter experiments were carried out in a greenhouse to study the fate of HTO following its soil deposition at different growth stages of Chinese cabbage. An HTO solution was applied to the surface of an acidic sandy soil at a time before, and four different times after, sowing. The transfer of HTO to the plants was quantified with the areal transfer factor (TFa, m2 kg(-1)-fresh) defined as the ratio of the plant concentration at harvest to the areal activity deposition. In the four post-sowing applications, the TFa values were in the ranges of 1.6 x 10( 5)-4.9 x 10(-3) for TFWT and 4.5 x 10(-6) -4.3 x 10(-5) for OBT, increasing with a decrease in the time interval between application and harvest. In the pre sowing application, which was followed by a soil mixing, the TFa values for TFWT and OBT were 1.3 x 10(-4) and 8.6 x 10(-6), respectively. One week after harvest, soil samplings were made for the applications at 26 (A(26)) and 63d (A(63)) after sowing. Peaks of the depth profiles of the soil HTO appeared in the 10-15 cm layer for A(26) and the 5-10 cm layer for A(63). The top 30 cm of soil contained 0.5% and 20% of the applied activity for A(26) and A(63), respectively. Negligible fractions seemed to be in the deeper zone. It was estimated that almost all or most of the applied HTO had escaped to the air before plants' harvest. PMID- 17418921 TI - Creep and the in vivo assessment of human patellar tendon mechanical properties. AB - BACKGROUND: Owing to the viscoelastic nature of tendons it may be that the total excursion and hence strain experienced by the tendon under load may be affected by the duration of contraction. Here we examine the effect of contraction duration on the measured in vivo mechanical properties of the patellar tendon. METHODS: Nine healthy young men aged 21 (SEM 0.5 years) performed three short (3 4s) and three long (10-12s) maximal ramped isometric contractions on an isokinetic dynamometer, with real-time recordings of patellar excursions using B mode ultrasonography synchronised with forces to determine tendon mechanical properties. FINDINGS: Maximal patellar excursion was approximately 42% (P<0.001, effect size (r)=0.9) lower for the short 3.6 (SEM 0.4mm) vs. the long 6.2 (SEM 0.4mm) contractions. Similarly, across the range of forces tested, strain was approximately 42% (P<0.001, r=0.9) lower for the short vs. the long contractions 4.5 (SEM 0.5) vs. 8.0 (SEM 0.9%), respectively. Tendon stiffness however, was approximately 77% greater (4648 SEM 434 vs. 2633 SEM 257 N mm(-1), P<0.001, r=0.9) for short vs. long contractions. INTERPRETATION: Contraction duration significantly affects tendon strain and associated measures of stiffness at all levels of force. The implications of this finding are twofold in that the results: (a) indicate that in order to compare tendon mechanical properties within or across studies, duration of contraction must be standardised and (b) have possible implications on training protocols and associated injury risks. PMID- 17418922 TI - Leg kinematics and kinetics in landing from a single-leg hop for distance. A comparison between dominant and non-dominant leg. AB - BACKGROUND: Anterior cruciate ligament (ACL) deficiency can be a major problem for athletes and subsequent reconstruction of the ACL may be indicated if a conservative regimen has failed. After ACL reconstruction signs of abnormality in the use of the leg remain for a long time. It is expected that the landing after a single-leg hop for distance (horizontal hop) might give insight in the differences in kinematics and kinetics between uninjured legs and ACL reconstructed legs. Before the ACL-reconstructed leg can be compared with the contralateral leg, knowledge of differences between legs of uninjured subjects is needed. METHODS: Kinematic and kinetic variables of both legs were measured with an optoelectronic system and a force plate and calculated by inverse dynamics. The dominant leg (the leg with biggest horizontal hop distance) and the contralateral leg of nine uninjured subjects were compared. FINDINGS: No significant differences were found in most of the kinematic and kinetic variables between dominant leg and contralateral leg of uninjured subjects. Only hop distance and hip extension angles differed significantly. INTERPRETATION: This study suggests that there are no important differences between dominant leg and contralateral leg in healthy subjects. As a consequence, the uninvolved leg of ACL-reconstructed patients can be used as a reference. The observed variables of this study can be used as a reference of normal values and normal differences between legs in healthy subjects. PMID- 17418923 TI - Effects of flow regime and flooding on heavy metal availability in sediment and soil of a dynamic river system. AB - The acid volatile sulphide (AVS) and simultaneously extracted metals (SigmaSEM) method is increasingly used for risk assessment of toxic metals. In this study, we assessed spatial and temporal variations of AVS and SigmaSEM in river sediments and floodplain soils, addressing influence of flow regime and flooding. Slow-flowing sites contained high organic matter and clay content, leading to anoxic conditions, and subsequent AVS formation and binding of metals. Seasonality affected these processes through temperature and oxygen concentration, leading to increased levels of AVS in summer at slow-flowing sites (max. 37micromolg(-1)). In contrast, fast-flowing sites hardly contained AVS, so that seasonality had no influence on these sites. Floodplain soils showed an opposite AVS seasonality because of preferential inundation and concomitant AVS formation in winter (max. 3-30micromolg(-1)). We conclude that in dynamic river systems, flow velocity is the key to understanding variability of AVS and SigmaSEM. PMID- 17418924 TI - The value of "life at any cost": talk about stopping kidney dialysis. AB - With the trend toward an older, sicker dialysis population in the USA, discussions of ethical issues surrounding dialysis have shifted from concerns about access to and availability of the therapy, to growing unease about non initiation and treatment discontinuation. Recent studies report treatment withdrawal as the leading cause of death among elderly dialysis patients. Yet, the actual activities that move patients toward stopping treatment often remain obscure, even to clinicians and patients themselves. This paper explores that paradox, drawing on anthropological research among patients over age 70, their families, and clinicians in two California renal dialysis units. It concludes that many older patients sacrifice a sense of choice about dialysis in the present to maintain "choice" as both value and possibility for the future. Even so, patients desire more information and communication, provided earlier in their illness, about prognosis, how long they can expect to be on dialysis, and what the impact of the treatment will be on their daily lives. That, with time, there is a transition to be made from dialysis as "treatment" to end of life care could be better explained and managed to alleviate patients' confusion and unneeded isolation. PMID- 17418925 TI - Suppressive effects of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression by Citrus reticulata extract in RAW 264.7 macrophage cells. AB - Immature peels of Citrus reticulata extract (CR) are widely used as traditional herbal medicine in Korea. We studied its effects on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in RAW 264.7 macrophage cells. NO production was assessed by nitrite assay and iNOS expression was identified by reverse transcription-polymerase chain reaction (RT-PCR), Real-time PCR and Western blot. The promoter activity of iNOS gene was also determined by luciferase reporter gene assay using 5'-flanking region of murine iNOS gene. Activation of nuclear factor kappa B (NF-kappaB) was determined by electrophoretic mobility shift assay (EMSA). CR (20, 50, and 100 microg/ml) significantly inhibited the lipopolysaccharide (LPS)-induced NO production (P<0.01; 9.2+/-1.5, 4.8+/-0.6, and 3.3+/-0.4 microM), iNOS protein (38.1+/-3.8, 32.3+/-5.8, and 36.8+/-4.5%) and mRNA expression (34.2+/-4.1, 13.1+/-5.8, and 20.8+/-1.2%) in RAW 264.7 macrophage cells. CR (20, 50, and 100 microg/ml) also reduced the iNOS promoter activity (68.7+/-3.9, 50.6+/-5.6, and 45.9+/-3.9%) in piNOS-LUC-transfected cells. In addition, CR (20, 50, and 100 microg/ml) significantly inhibited the activity of NF-kappaB DNA binding activity in LPS induced macrophage cells (P<0.05; 51.8+/-4.1, 32.7+/-5.5, and 35.7+/-2.9%). These results suggest that CR may suppress LPS-stimulated NO production by inhibiting of NF-kappaB. PMID- 17418926 TI - Acute toxicity of sodium arsenite in a complex food matrix. AB - Acute toxicity of a single oral dose of sodium arsenite (As), administered in half and half cream (HH), was assessed in male and non-pregnant female rats (0.41, 4.1, 41.0 and 410.0mg/kg body weight) and pregnant rats (0.41, 4.1 and 41.0mg/kg body weight). Control rats received deionized water alone, HH alone or 41.0mg/kg As in deionized water (41 mg/kg As-water). Male and non-pregnant rats were monitored for 14 consecutive days post-dosing. Pregnant rats, dosed on gestation day 10 (GD-10), were monitored until fetuses were collected on GD 20. High mortality (100%) was observed in male and non-pregnant female rats exposed to 410.0mg/kg As-HH. Low mortality (25%) was observed in non-pregnant female rats exposed to 41 mg/kg As-water. No mortality was observed in other control or treated groups. Reduced female fetal numbers were observed in the 41 mg/kg As water group but not in the other control groups. Developmental effects were not observed in the controls or the As-HH treatment groups. In conclusion, As toxicity was not reduced when a high dose (410 mg/kg) was administered in HH however, at lower doses (41 mg/kg), HH reduced acute As oral toxicity in the female and developing fetus. PMID- 17418927 TI - Content and daily intake of copper, zinc, lead, cadmium, and mercury from dietary supplements in Mexico. AB - This study investigates the presence of Cu, Zn, Cd, Pb, and Hg in 24 dietary supplements purchased in different health stores across the city of Hermosillo, located in the northwest of Mexico. Analysis of metals was done by microwave digestion and atomic absorption spectroscopy. The most abundant elements in dietary supplements were Cu (<0.19-137.85 microg/g) and Zn (<2.83-4785.71 microg/g), followed by Pb (<0.003-66.32 microg/g), Cd (<0.001-2.90 microg/g), and Hg (<0.24-0.85 microg/g). The estimated daily intakes of metals were below those recommended by WHO and the Institute of Medicine, showing that little risk from heavy metals is associated with the consumption of the dietary supplements analyzed. However, some products presented more than 10% of the tolerable daily intake of Pb, indicating that production processes should be improved. PMID- 17418928 TI - A 90-day ad libitum administration toxicity study of oligoglucosamine in F344 rats. AB - A 90-day ad libitum administration toxicity study of oligoglucosamine (OG) was carried out using F344 rats of both sexes. The animals were divided into four groups of 20 animals each, 10 of each sex, and fed a diet containing 0, 0.04, 0.2 or 1.0 (w/w)% OG. During the administration period, no animals of either sex died or exhibited abnormal signs in the 0.04% OG and 0.2% OG groups. In the 1% OG group, in both sexes, erythema and swelling of the snout and forelimbs and loss of fur in the forelimbs were observed. On macroscopic observation, emaciation, swelling of the snout, auricles and forelimbs and alopecia of the forelimbs were also observed in 2-3 males of the 1% OG group. It was suggested that these topical abnormalities might be due to dermal responses to OG adhering to the skin and fur, which are easily soiled with saliva during grooming. In the animals of the 1% OG group, food consumption decreased, resulting in body weight gain being suppressed. This was found concomitantly with the abnormal findings mentioned above. Thus, feeding difficulties due to the topical lesions on the snout and forelimbs were thought to affect body weight. In hematology, platelet count, lymphocyte count and differential neutrophil count increased in males of the 1% OG group. These changes might be related to the dermal inflammation. Abnormalities in urinalysis and blood chemistry, as well as a small thymus, small spleen, dark spots or areas on the glandular stomach mucosa, pale Harderian glands and small testes in histopathology, were also observed in males in the 1% OG group. Whether or not all these changes were related only to the malnutrition remains to be elucidated. From these results, OG gave rise to no adverse effects in rats up to the dose level of 0.2 (w/w)%. Thus, the no observed adverse effect level was determined to be 0.2 (w/w)% for rats of either sex (124.0mg/kg/day in males, 142.0mg/kg/day in females). PMID- 17418929 TI - [Extra-abdominal desmoid tumor in a scar of donor-site of a latissimus dorsi flap: case report]. AB - Tumefaction arising lately after latissimus dorsi flap harvest are rare and observed in 1 or 2% of the cases. These lesions are frequently related to kystic sero-hematoma and are easily and efficiently treated with surgical excision. In some rare circumstances, a tumoral evolution can mimic a kystic sero-hematoma. We will discuss one case of desmoid tumor arising from a latissimus dorsi flap donor site scar. The subject was a 45 years old woman who had a breast reconstruction following mastectomy. A dorsal tumefaction, with a benign aspect, was observed during the follow-up period. The biopsy showed an extra-abdominal desmoid tumor. The patient was treated with a large excision of the lesion and reconstructed using two opposing local cutaneous advancing flaps. No radicalization was necessary. No sign of recurrence has been observed at 4 years follow-up. PMID- 17418930 TI - "DNA, more than just a ladder". A tribute to Michel Duguet (1943-2005). PMID- 17418931 TI - Transport viewed as a catalytic process. AB - Transport catalysis is analysed in terms of the "induced transition fit" (ITF) concept. The essentials of ITF are briefly elucidated, emphasizing the difference of substrate-protein interactions between enzymes and carriers exemplified by the paradigm ADP/ATP carrier (AAC). Two of the numerous applications of the ITF are discussed in more detail: unidirectional passive and active transport and the relation of substrate site type inhibitors to the carrier conformations. According to ITF in most cases of unidirectional transport intrinsic binding energies may be insufficient for transport catalysis and requires additional energy from ATP or electrochemical gradients. The impacts of external energy on the carrier cycle are examined for ABC transporters (mdr) and for cation substrate co-transporters (LacY). The relations of inhibitors to the binding site of the carrier are discussed, given the paradigm examples of side specific inhibitors of the AAC. Results with the AAC suggest the induction of an abortive ground state by inhibitors, representing extreme side specific conformation of the binding center. PMID- 17418932 TI - Inhibition of peroxynitrite precursors, NO and O2, at the onset of reperfusion improves myocardial recovery. AB - AIM OF STUDY: Previous reports note an increase in both reactive oxygen species (ROS) and nitric oxide (*NO) at the onset of myocardial reperfusion. We tested the hypothesis that inhibition of *NO or ROS production at the time of reperfusion improves recovery of post-ischemic myocardial function. METHODS AND MATERIALS: Isolated rat hearts were perfused with temperature controlled (37.4 degrees C) modified Krebs Henseleit buffer solution at 85 mm Hg. Following 20 min of global ischemia, hearts were reperfused for the first 10 min with: (1) standard buffer (control), (2) buffer with a NOS inhibitor, N-nitro-L-arginine methyl ester (L-NAME), (3) buffer with superoxide dismutase (SOD) or (4) buffer with N-morpholinosydnonimine hydrochloride (SIN-1), a peroxynitrite generator. Tissue O(2) and *NO were continuously measured with thin electrochemical probes embedded in the wall of the LV. ROS was measured with the spin trap 5,5-dimethyl 1-pyrroline N-oxide (DMPO) (40 mM). LV contractile function was continuously monitored. RESULTS: Recovery of LV contractile function was significantly improved in hearts initially reperfused with L-NAME and SOD and significantly depressed in hearts reperfused with SIN-1 compared with control (p<0.01, n=5-8 per group). DMPO-adduct during reperfusion (measure of ROS) was significantly decreased with SOD (p<0.001 versus L-NAME and Control, n=4 per group) and unchanged with L-NAME and SIN-1 compared with Control. With L-NAME, tissue *NO and PO(2) were significantly decreased, independent of coronary flow, during reperfusion compared with control and SIN-1. CONCLUSIONS: Inhibition of O(2)*(-) or *NO at the time of reperfusion improves early reperfusion LV function and alters tissue oxygen tension. In contrast to pre-ischemic treatments, intervention to reduce peroxynitrite generation at the onset of reperfusion can effectively improve post-ischemic myocardial recovery. PMID- 17418933 TI - Protection strategies for drinking groundwater sources in small Quebec municipalities. AB - Awareness of groundwater protection has increased substantially in recent decades. In the Province of Quebec, Canada, the Groundwater Catchment Regulation (GWCR) was promulgated in 2002 to protect water quality in public wells. The goal of the present study was to document groundwater protection in the context of emerging regulations and identify factors explaining the propensity of municipalities applying protection strategies. Two types of information were used in this study: data from a questionnaire-based survey conducted among 665 municipalities in the Province of Quebec and complementary information gathered from various sources. Data from the survey revealed that fewer than half of the municipalities have been able to comply with the GWCR, mainly because of financial limitations. Also, close to half of the municipalities have either identified or are expecting land use conflicts to arise between protection areas required by the GWCR and other land usage, with agriculture being the main conflicting activity. Multivariate logistic regression models served to identify factors explaining the likelihood of municipalities to take groundwater protection measures. Those factors were municipality revenue, history of water contamination in distribution systems, land use near wellheads, location of municipalities within a provincial priority watershed and the importance of groundwater use in a region. Results of the study may prove helpful for government authorities in better understanding the groundwater protection issue and in implementing strategies that improve the ability of municipalities to protect groundwater. PMID- 17418934 TI - Evaluating sustainable forest management strategies with the Analytic Network Process in a Pressure-State-Response framework. AB - Nowadays forestry faces a complex management situation; the understanding of sustainable forest management (SFM) has gone far beyond the original meaning of sustainable yield of timber. SFM strategies should fulfil ecological, economic and social functions without causing damage to other ecosystems. In this understanding, forest management actions cannot be seen as isolated or mono causal. In this case study, indicators for SFM are arranged in a Pressure-State Response (PSR) framework at forest management unit level. This framework links pressures on the environment caused by human activities with changes of environmental state (condition) parameters. Forest management also responds to these changes by instituting environmental and economic measures to reduce pressures and restore natural resources. The Analytic Network Process (ANP) is utilized to evaluate the performance of four management strategies with regard to the PSR framework on SFM. Priorities of indicators and alternatives are modelled with the ANP resulting from the interconnections to other indicators and their respective cumulative importance. The approach allows for more detailed information on the network of human influences and their impacts on forest ecosystems and goes beyond the limitations of flat-dimensioned indicator sets. PMID- 17418935 TI - Editorial comment on: Outcome of prostate cancer patients with initial PSA> or =20 ng/ml undergoing radical prostatectomy. PMID- 17418936 TI - Preserved postoperative penile size correlates well with maintained erectile function after bilateral nerve-sparing radical retropubic prostatectomy. AB - OBJECTIVES: Controversial data on penile length after radical retropubic prostatectomy are available. We hypothesised that postoperative penile size correlates to erectile function following bilateral nerve-sparing radical retropubic prostatectomy (BNSRRP). METHODS: Thirty-three consecutive patients with a preoperative erectile function domain of the International Index of Erectile Function (IIEF-EF) score indicating full potency (> or = 26) were prospectively enrolled. All patients underwent BNSRRP performed by one high volume surgeon. All patients were preoperatively evaluated by IIEF-EF, analysis of comorbidities, physical examination, and penile power colour Doppler ultrasound using intracavernosal injection of prostaglandin E(1) (PGE(1)) 20 microg plus audiovisual and manual genital stimulation. Penile length and circumference were measured in flaccidity and at maximum erection. Six months postoperative, patients were assessed with the same protocol plus general assessment questions investigating penile structure and function. Statistical analysis was performed with an independent sample t test. RESULTS: Mean patient age was 56.5 yr. We found no difference between the preoperative and the 6-mo postoperative mean IIEF-EF domain score (27.2 vs, 26.7, respectively; p = 0.35). No difference was found in penile colour Doppler evaluation between the preoperative and postoperative periods (all p values > or = 0.3). We found no differences in penile length and circumference between the preoperative and postoperative evaluation either in the flaccid or in the erect state. Mean flaccid penile length (cm; preop vs. postop): 13.2 vs. 13 (p = 0.6). Mean flaccid penile circumference (cm; preop vs. postop): 11.1 vs. 11 (p = 0.7). Mean erect penile length (cm; preop vs. postop): 16.8 vs. 16.5 (p = 0.08). Mean erect penile circumference (cm; preop vs. postop): 15.6 vs. 15.3 (p = 0.2). CONCLUSIONS: This is the first report on penile changes in flaccidity and at maximum erection after BNSRRP in patients treated by one high-volume surgeon. The postoperative preservation of erectile function positively correlated with the maintenance of penile length following surgery. We found no change in penile size after surgery. PMID- 17418938 TI - Outcome of prostate cancer patients with initial PSA> or =20 ng/ml undergoing radical prostatectomy. AB - OBJECTIVES: To retrospectively assess the outcome of patients with initial PSA of 20 ng/ml or higher undergoing radical prostatectomy (RP) for prostate cancer (pCA). METHODS: Between January 1986 and June 2005, 275 patients with preoperative PSA> or =20 ng/ml underwent RP for pCA at our institution. Overall, disease-specific and biochemical progression-free survival rates for the entire cohort and for particular subgroups were determined. RESULTS: Median patient age at time of surgery was 64 yr (range: 44-75). Fifty-seven patients (20.7%) had pT2 stage, 206 (74.9%) pT3, and 10 (3.7%) pT4; 78 (28.4%) presented with local nodal metastases (pN+). To date, 40 patients have died (14.5%), 22 of pCA and 18 of other causes. Biochemical progression occurred in 92 patients (33.5%). Overall (and disease-specific) survivals at 5, 10, and 15 yr were 87% (93%), 70% (83%), and 58% (71%), respectively. These survival rates did not significantly differ between patients receiving immediate versus deferred hormonal therapy (in case of progression). Five-year PSA progression-free survival in patients on surveillance (receiving deferred hormonal treatment at the onset of rising PSA values) was 53%. For patients on immediate hormonal treatment following RP, the 5-yr hormone refractory PSA progression rate was 76%. CONCLUSIONS: According to long-term follow-up results in this high-risk cohort of patients with preoperative PSA> or =20 ng/ml, RP can be considered a viable therapeutic option. With regard to combining immediate hormonal therapy with surgery, the optimal treatment following RP remains to be defined. PMID- 17418939 TI - Changes in the fluctuation of the contraction rhythm of spontaneously beating cardiac myocytes in cultures with and without cardiac fibroblasts. AB - The heart functions as a syncytium of cardiac myocytes and surrounding supportive non-myocytes such as fibroblasts. There is a possibility that a variety of non myocyte-derived factors affect the maturation of cardiac myocytes in the development of the heart. Cultured neonatal cardiac myocytes contract spontaneously and cyclically. The fluctuation of beating rhythm varies depending on the strength of coupling through gap junctions among cardiac myocytes, indicating that the development of intercellular communication via gap junctions is crucial to the stability of contraction rhythm in cardiac myocytes. In this study, we aimed at elucidating whether and how cardiac fibroblasts affect the development of cardiac myocytes from the point of view of the changes in the fluctuation of the contraction rhythm of cardiac myocytes in cardiac myocyte fibroblast co-cultures. The present study suggested that cardiac fibroblasts co cultured with cardiac myocytes enhanced the intercellular communication among myocytes via gap junctions, thereby stabilizing the spontaneous contraction rhythm of cultured cardiac myocytes. PMID- 17418940 TI - A P system and a constructive membrane-inspired DNA algorithm for solving the Maximum Clique Problem. AB - We present a P system with replicated rewriting to solve the Maximum Clique Problem for a graph. Strings representing cliques are built gradually. This involves the use of inhibitors that control the space of all generated solutions to the problem. Calculating the maximum clique for a graph is a highly relevant issue not only on purely computational grounds, but also because of its relationship to fundamental problems in genomics. We propose to implement the designed P system by means of a DNA algorithm. This algorithm is then compared with two standard papers that addressed the same problem and its DNA implementation in the past. This comparison is carried out on the basis of a series of computational and physical parameters. Our solution features a significantly lower cost in terms of time, the number and size of strands, as well as the simplicity of the biological implementation. PMID- 17418941 TI - Teaching evidence-based clinical practice to neurology and neurosurgery residents. AB - The primary objective of education in evidence-based clinical practice (EBCP) is to provide a resident or student with the requisite skills to satisfactorily solve real everyday clinical problems throughout their careers and to translate those solutions into better care for patients. At the University of Western Ontario in London, Canada, there is a well-developed and highly successful evidence-based neurology curriculum primarily aimed at residents. This article summarizes the current context of EBCP postgraduate training in the neurological sciences and a detailed description of its purpose, learning outcomes, required resources, content, teaching strategies, and assessment tools. In this program, we teach the principles of EBCP through the review of pertinent neurological clinical questions. The summary of each topic is recorded on our website in the form of critically appraised topics (CATs) in electronically accessible CAT banks. PMID- 17418942 TI - Hazmat transport: a methodological framework for the risk analysis of marshalling yards. AB - A methodological framework was outlined for the comprehensive risk assessment of marshalling yards in the context of quantified area risk analysis. Three accident typologies were considered for yards: (i) "in-transit-accident-induced" releases; (ii) "shunting-accident-induced" spills; and (iii) "non-accident-induced" leaks. A specific methodology was developed for the assessment of expected release frequencies and equivalent release diameters, based on the application of HazOp and Fault Tree techniques to reference schemes defined for the more common types of railcar vessels used for "hazmat" transportation. The approach was applied to the assessment of an extended case-study. The results evidenced that "non accident-induced" leaks in marshalling yards represent an important contribution to the overall risk associated to these zones. Furthermore, the results confirmed the considerable role of these fixed installations to the overall risk associated to "hazmat" transportation. PMID- 17418943 TI - Chemical compatibility of PU/PAN interpenetrating polymer network membrane with substituted aromatic solvents. AB - Polyethylene glycol (PEG)-based polyurethane/polyacrylonitrile (PU/PAN, 50/50) semi-interpenetrating polymer network (SIPN) membrane has been studied from sorption/desorption cycles and diffusion behaviour with substituted aromatic probe molecules at 20, 40 and 60 degrees C. Sorption/desorption cycles have been repeated to evaluate polymer-solvent interaction. Organic solvents taken up or given out by IPN are measured periodically till equilibrium. Using these data, sorption (S), diffusion (D) and permeation (P) coefficients have been calculated from Fick's equation. Sorption data is correlated with solubility parameter of solvents and polymer. It was found that solvents of comparable solubility parameter with IPN interact more and thus there is an increase in sorption. Molecular mass between cross-link has been calculated using Flory Rehner equation. The cross-link density and degree of cross-linking of the membrane is calculated. From the temperature dependence of sorption and diffusion coefficients, the Arrhenius activation parameters like activation energy for diffusion (E(D)) and permeation (E(P)) processes have been calculated. Furthermore, the sorption results have been interpreted in terms of thermodynamic parameters such as change in enthalpy (DeltaH) and entropy (DeltaS). Concentration profiles of penetrants at different penetration depths in the polymer sample at different time intervals have also been calculated theoretically from a solution of Fick's equation under appropriate initial boundary conditions. PMID- 17418944 TI - Abcisic acid and gibberellic acid cause increased lipid peroxidation and fluctuated antioxidant defense systems of various tissues in rats. AB - The study was aimed at demonstrating changes in the antioxidant defense systems [Reduced glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase (CAT)] forming an antioxidative barrier and oxidative stress parameter (Malondialdehyde=MDA) in the various tissues of Sprague-Dawley rats which were administrated plant growth regulators (PGRs) [Abcisic acid (ABA) and Gibberellic acid (GA(3))] during 25 days. Seventy five parts per million of ABA and GA(3) as drinking water were administered orally to rats (Sprague-Dawley albino) ad libitum for 25 days continuously. The PGRs treatments caused different effects on antioxidant defense systems and MDA content of experimented rats compared to controls. The lipid peroxidation end product MDA significantly increased in the spleen and lungs of rats treated with ABA and GA(3) without significantly change in the other tissues. The GSH levels were significantly increased in the lungs and stomach of rats treated with ABA without any change in the tissues of rats treated with GA(3). Antioxidant enzyme activities such as SOD significantly increased in the spleen of rats treated with ABA and GA(3). Meanwhile, SOD significantly increased in the kidney of rats treated with GA. CAT significantly decreased in the lungs treated with ABA but did not change significantly in all the rest of rat tissues treated with both the PGRs. On the other hand, the ancillary enzyme GR activity decreased in the spleen and increased in the kidney with GA(3) treatment. The drug metabolizing enzyme GST activity significantly decreased in the heart of rats treated with GA(3) but increased in the spleen and lungs of rats treated with both PGRs. The observations presented led us to conclude that administration of subacute ABA and GA(3) promotes lipid peroxidatin content and alters in the antioxidative systems in the rat's various tissues. These data, along with changes, suggest that the PGRs produced oxidative stress in rats during the period of a 25-day subchronic exposure. PMID- 17418945 TI - Expression of TrkA receptor for neurotrophins in trigeminal neurons innervating the rat gingivomucosal tissue. AB - The purpose of this study was to characterize and evaluate the expression of TrkA receptor in trigeminal ganglion (TG) neurons that innervate the rat gingivomucosal tissue. A retrograde nerve tracer Fluorogold (FG) was injected into the gingiva (group 1) or applied into the gingival sulcus (group 2) of the first right maxillary molar to identify the neurons in TG that innervate the gingivomucosa. After 10 days TG were dissected and FG fluorescence in neurons was observed under UV light microscope. To draw a comparison, approximately 1000 neurons per ganglion from the entire TG (group 3) and approximately 350 neurons per ganglion from the maxillary region in TG (group 4), were analyzed. Expression of TrkA receptor in TG neurons was investigated by immunohistochemistry. About 70% of neurons in groups 1 and 2 contained TrkA receptor, which was statistically significantly more than in groups 3 (41%) and 4 (38%). FG-labeled TrkA immunopositive neurons were predominantly small or medium-sized (less than 1200microm(2)). However, the neurons innervating the rat gingivomucosa were on average larger than the neurons in the entire TG or in the maxillary region. In conclusion, the majority of neurons in TG that innervate the rat gingivomucosa are small or medium-sized, contain TrkA receptor and are most probably nociceptive. PMID- 17418946 TI - A novel adenoviral vector-mediated neuronal selective gene expression in neonatal mouse brain in response to hypoxia. AB - Selective gene expression targeting neurons is a challenge, which, if successfully overcome, carries an enormous potential for clinical applications in therapeutics against neurodegenerative diseases. We have reported previously the construction of a series of adenoviral vectors capable of selectively expressing a reporter gene luciferase in cultured neurons [D. Huang, A. Desbois, S.T. Hou, A novel adenoviral vector which mediates hypoxia-inducible gene expression selectively in neurons, Gene Ther. 12 (2005) 1369-1376]. A combination of neuron restrictive silencer elements (NRSEs), hypoxia responsive elements (HREs) and CMV minimal promoter (CMVmp) was packaged into replication defective adenovirus to target gene expression selectively in neurons in a hypoxia-regulated manner. In the present study, we injected the adenoviral vectors into the neonatal mouse brain followed by treatment with hypoxia. The expression of the reporter luciferase gene was examined by luciferase assay and fluorescent immunostaining. Neurons and glial cells were identified by staining with antibodies against NeuN and GFAP, respectively. Remarkably, in response to hypoxia, Ad/5HRE-3NRSE showed strong hypoxia-inducible gene expression of the reporter luciferase selectively in neurons but not in glial cells. In contrast, brains infected with the control vector Ad/5HRE showed no selectivity in luciferase expression (in both neurons and glial cells) under the hypoxic condition. Taken together, these studies demonstrated that this vector (Ad/5HRE-3NRSE) can mediate gene expression selectively in neurons both in vitro and in vivo, supporting the suggestion that further refinement of this vector may lead to the development of a useful tool to investigate mechanisms of neuronal damage following cerebral ischemia and a possible effective gene therapy vector to stroke. PMID- 17418947 TI - Mutant SOD1G93A in bone marrow-derived cells exacerbates 3-nitropropionic acid induced striatal damage in mice. AB - 3-Nitropropionic acid (3-NP), an irreversible inhibitor of succinate dehydrogenase, produces selective lesions in striatal neurons that resemble those observed in Huntington's disease neuropathology. In this study, we evaluated the role of peripheral bone marrow-derived cells (BMDCs) in the 3-NP-induced striatal damage by transplanting bone marrow cells with human SOD1 G93A mutation (mSOD1(G93A)) which induces amyotrophic lateral sclerosis through an unknown gain of toxicity and mitochondrial dysfunction. We assessed striatal damage after 3-NP treatment in the recipient C57BL/6 wild-type (WT) mice that received bone marrow cells from WT or mSOD1(G93A) transgenic donor mice (WT-->WT or mSOD(G93A)-->WT). After intraperitoneal injection of 3-NP, six of the eight mSOD1(G93A)-->WT mice had bilateral striatal lesions while only one out of eight WT-->WT mice had a striatal lesion. The lesion volume was significantly higher in the mSOD1(G93A)- >WT mice than in the WT-->WT mice. However, following an intrastriatal injection of 3-NP, there was no significant difference in the lesion volumes between the WT ->WT mice and mSOD1(G93A)-->WT mice. Thus, the exacerbation of 3-NP-induced striatal damage in mSOD(G93A)-->WT mice was only seen after systemic administration of 3-NP, but not after intrastriatal injection. These results demonstrate that altered SOD1 activity (mSOD(G93A)) in BMDCs affects striatal damage probably through a mechanism involving a systemic factor. PMID- 17418948 TI - IL-15 and IL-15R alpha gene deletion: effects on T lymphocyte trafficking and the microglial and neuronal responses to facial nerve axotomy. AB - IL-15 is a potent T cell chemoattractant, and this cytokine and its unique alpha subunits, IL-15R alpha, can modify immune cell expression of several T cell chemokines and their receptors. Facial nerve axotomy in mice leads to T cell migration across an intact blood-brain-barrier (BBB), and under certain conditions T cells can provide neuroprotection to injured neurons in the facial motor nucleus (FMN). Although chemokines and chemoattractant cytokines are thought to be responsible for T cell migration to the injured cell bodies, data addressing this question are lacking. This study tested the hypothesis that T cell homing to the axotomized FMN would be impaired in knockout (KO) mice with the IL-15 and IL-15R alpha genes deleted, and sought to determine if microglial responsiveness and motoneuron death are affected. Both IL-15KO and IL-15R alpha KO mice exhibited a marked reduction in CD3(+) T cells and had fewer MHC2(+) activated microglia in the injured FMN than their respective WT controls at day 14 post-axotomy. Although there was a relative absence of T cell recruitment into the axotomized FMN in both knockout strains, IL-15R alpha KO mice had five times more motoneuron death (characterized by perineuronal microglial clusters engulfing dead motoneurons) than their WT controls, whereas dead neurons in IL 15KO did not differ from their WT controls. Further studies are needed to dissect the mechanisms that underlie these observations (e.g., central vs. peripheral immune contributions). PMID- 17418949 TI - Granulomatous nephritis in psittacines associated with parasitism by the trematode Paratanaisia spp. AB - Trematodes belonging to the family Eucotylidae are parasites of the kidney and ureter, and affect several bird species. However, psittacines have not been identified as hosts of these parasites. Three birds, an adult female blue and gold macaw (Ara ararauna), an adult female blue-winged macaw (Propyrrhura maracana) and an adult male white-eared parakeet (Pyrrhura leucotis) were admitted at the Veterinary Hospital of the Fundacao Zoo-Botanica de Belo Horizonte, Brazil (FZB/BH). All three birds had severe dehydration and cachexia. The blue and gold macaw presented with dyspnea, apathy, and incoordination. Blood cell counts indicated discrete anemia and leucopenia. Blood biochemistry revealed significant increase in levels of uric acid (61 mg/dl) and blood urea nitrogen (22 mg/dl). The bird died within 24 h after admission. The other two birds were admitted with similar clinical signs, but died prior to a complete clinical examination. At the necropsy, in all the three birds, the kidneys were enlarged with brown-yellowish discoloration and irregular cortical surface. On the cut surface, there was a brown-yellowish material with few visible parasites flowing out of the parenchyma. When fragments of the kidneys were placed in 10% formalin, a large number of trematodes came out of the renal parenchyma. The parasites were identified as Paratanaisia robusta infecting all three birds, and P. bragai infecting the blue-winged macaw and the white-eared parakeet. Histologically, there was an interstitial, multifocal to coalescent, lymphoplasmacytic infiltrate with some epithelioid macrophages, and a few heterophils, characterizing a granulomatous nephritis. Adult worms and eggs were observed within dilated tubules and in the renal pelvis. In the blue and gold macaw, some parasite eggs were located interstitially associated with an intense adjacent granulomatous reaction. PMID- 17418950 TI - Seroprevalence and Toxoplasma gondii-IgG avidity in sheep from Lajes, Brazil. AB - Sheep are important domestic animals in the Northeast region of Brazil due to their minimal rearing and maintenance costs, and to their production of both meat and milk. In animals, Toxoplasma gondii infection results in significant reproductive and economic losses. The epidemiology of toxoplasmosis in sheep in the Northeast of Brazil has been little studied; particularly in Rio Grande do Norte State. Sera from 102 sheep intended for consumption in Lajes were subjected to the Toxoplasma-ELISA test to detect anti-T. gondii specific IgG confirming a past infection. Of the total tested, 30 (29.41%) sera were positive for IgG with an increasing number of positive animals with advancing age. We used IgG avidity ELISA in 30 positive samples and observed that 6 (20%) had low avidity antibodies and 24 (80%) had high avidity antibodies. Epidemiological studies are required in order to identify sources of infection for these hosts as well as their impact on animal breeding in the region and risk of transmission to humans. PMID- 17418951 TI - Application of conventional and real-time fluorescent ITS1 rDNA PCR for detection of Besnoitia besnoiti infections in bovine skin biopsies. AB - Besnoitia besnoiti, an apicomplexan protozoan parasite, is the causative agent of bovine besnoitiosis. This infection may dramatically affect body condition, and, in males, lead to irreversible infertility. While identification of clinical cases and their histopathological confirmation is relatively simple to carry out, finding subclinical forms of infection is more difficult, thus a more sensitive test for the identification of the etiological agent may be an appropriate diagnostic tool. We have developed the ITS1 rDNA-sequence-based conventional and real-time PCR which are highly sensitive and specific for the detection of B. besnoiti infection in cattle. A recombinant internal positive control was introduced to assess possible sample-related inhibitory effects during the amplification reaction and, in order to prevent false-positive results, a pre-PCR treatment of potentially contaminating dU-containing PCR product with uracil-DNA glycosylase (UDG) was followed. PMID- 17418952 TI - A comparison of the efficacy of two ivermectin formulations against larval and adult Ascaris suum and Oesophagostomum dentatum in experimentally infected pigs. AB - A study was conducted to evaluate and compare the efficacy of two injectable formulations of ivermectin (IVM-1 and IVM-2) at a dose rate of 0.3 mg/kg bodyweight versus placebo in the treatment and control of larval and adult stages of Ascaris suum and Oesophagostomum spp. in experimentally infected pigs. Seventy helminth free pigs were allocated on a liveweight basis to 7 groups each comprising 10 pigs (A-G). Group A served as an untreated control group. Groups B and C were used to investigate the efficacy of both formulations against adult stages of A. suum and Oesophagostomum spp., Groups D and E for efficacy against larval stages of A. suum and Groups F and G for efficacy against larval stages of Oesophagostomum spp. Pigs of groups A, B, C, D and E were infected on Day-0 with 1000 infective A. suum eggs each. Infective larvae of Oesophagostomum spp. (10,000/pig) were given on Day-0 to pigs of Groups F and G and on Day-21 to pigs of Groups A, B and C. Treatment was given to pigs of Group A (saline as placebo) on Day-7 and -28, IVM-1 to pigs of Group F on Day-7, pigs of Group D on Day-14 and pigs of Group B on Day-49. IVM-2 was given to pigs of Group G on Day-7, Group E on Day-28 and Group C on Day-49. Pigs of Groups F and G were sacrificed on Day 28, pigs of Groups A, D and E on Day-49 and pigs of Groups B and C on Day-56. Post mortem worm counts showed the following efficacies: (IVM-1) against larval A. suum 100%, against adult A. suum 94.4%, against larval Oesophagostomum spp. 52.0% and against adult Oesophagostomum spp. 83.0%. (IVM-2) against larval A. suum 100%, against adult A. suum 90.3%, against larval Oesophagostomum spp. 94.0% and against adult Oesophagostomum spp. 94.7%. PMID- 17418954 TI - Inhibition of proliferation of Pseudomonas aeruginosa by KGF in an experimental burn model using human cultured keratinocytes. AB - Experimental models showed the ability of Pseudomonas aeruginosa to interact with epidermal keratinocytes [Green H, Kehinde O, Thomas J. Growth of cultured human epidermal cells into multiple epithelia suitable for grafting. Proc Natl Acad Sci USA 1979;76(11):5665-8], stimulating these cells to produce specific peptides that start an immunological chain reaction in the epidermis [O'Connor NE, Mulliken JB, Banks-Schlegel S, Kehinde O, Green H. Grafting of burns with cultured epithelium prepared from autologous epidermal cells. Lancet 1981;(1):75 8]. The immune reaction causes the release of cytokines and growth factors. The objective of this study was to test whether the presence of keratinocyte growth factor (KGF) alters P. aeruginosa proliferation in an experimental burn model. METHODS: Human keratinocytes derived from neonatal foreskins were isolated and cultured following standard methods [Gallico III, GG, O'Connor NE, Compton CC, Kehinde O, Green H. Permanent coverage of large burn wounds with autologous cultured human epithelium. N Engl J Med 1984;311(7):448-51]. Some of these cells were genetically modified to produce KGF, and the other cells were supplemented with different KGF concentrations in the culture media. Both groups of keratinocytes were seeded in collagen matrices and cultured to form stratified epithelia. A hot plate was used to produce burn defects. Each matrix was inoculated with luminescent P. aeruginosa strain. Experiments were made using keratinocytes without KGF, keratinocytes supplemented with different concentrations of KGF, and keratinocytes genetically modified to produce KGF. Statistical analyses were made using Wilcoxon paired test. RESULTS: When KGF was added to P. aeruginosa in the presence of keratinocytes, bacterial growth was inhibited, and the same was observed when genetically modified keratinocytes were used. CONCLUSION: Many studies have been done on KGF, where its known properties were defined as a mitogen for keratinocytes [Munster AM. Cultured skin for massive burns: a prospective, controlled trial. Ann Surg 1996;224(3):372-7]. This method allows for a qualitative and quantitative evaluation in real time of the bacterial growth in wound sites after bacterial inoculation. KGF was involved in the reduction of bacterial viability. However, as KGF alone did not produce any effect on P. aeruginosa, it seems to modulate the skin innate immune response. PMID- 17418955 TI - Group II metabotropic glutamate receptors inhibit glutamate release at thalamocortical synapses in the developing somatosensory cortex. AB - Thalamocortical synapses provide a strong glutamatergic excitation to cortical neurons that is critical for processing sensory information. Unit recordings in vivo indicate that metabotropic glutamate receptors (mGluRs) reduce the effect of thalamocortical input on cortical circuits. However, it is not known whether this reduction is due to a reduction in glutamate release from thalamocortical terminals or from a decrease in cortical neuron excitability. To directly determine whether mGluRs act as autoreceptors on thalamocortical terminals, we examined the effect of mGluR agonists on thalamocortical synapses in slices. Thalamocortical excitatory postsynaptic currents (EPSCs) were recorded in layer IV cortical neurons in developing mouse brain slices. The activation of group II mGluRs with (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG IV) reduced thalamocortical EPSCs in both excitatory and inhibitory neurons, while the stimulation of group I or group III mGluRs had no effect on thalamocortical EPSCs. Consistent with a reduction in glutamate release, DCG IV increased the paired pulse ratio and the coefficient of variation of the EPSCs. The reduction induced by DCG IV was reversed by the group II mGluR antagonist, LY341495, and mimicked by another selective group II agonist, (2R,4R)-4-aminopyrrolidine-2,4 dicarboxylic acid (APDC). The mGluR2 subtype appears to mediate the reduction of thalamocortical EPSCs, since the selective mGluR3 agonist, N acetylaspartylglutamate (NAAG), had no effect on the EPSCs. Consistent with this, we showed that mGluR2 is expressed in the barrels. Furthermore, blocking group II mGluRs with LY341495 reduced the synaptic depression induced by a short stimulus train, indicating that synaptically released glutamate activates these receptors. These results indicate that group II mGluRs modulate thalamocortical processing by inhibiting glutamate release from thalamocortical synapses. This inhibition provides a feedback mechanism for preventing excessive excitation of cortical neurons that could play a role in the plasticity and refinement of thalamocortical connections during this early developmental period. PMID- 17418956 TI - Variability in neuronal activity in primate cortex during working memory tasks. AB - Persistent elevated neuronal activity has been identified as the neuronal correlate of working memory. It is generally assumed in the literature and in computational and theoretical models of working memory that memory-cell activity is stable and replicable; however, this assumption may be an artifact of the averaging of data collected across trials, and needs experimental verification. In this study, we introduce a classification scheme to characterize the firing frequency trends of cells recorded from the cortex of monkeys during performance of working memory tasks. We examine the frequency statistics and variability of firing during baseline and memory periods. We also study the behavior of cells on individual trials and across trials, and explore the stability of cellular firing during the memory period. We find that cells from different firing-trend classes possess markedly different statistics. We also find that individual cells show substantial variability in their firing behavior across trials, and that firing frequency also varies markedly over the course of a single trial. Finally, the average frequency distribution is wider, the magnitude of the frequency increases from baseline to memory smaller, and the magnitude of frequency decreases larger than is generally assumed. These results may serve as a guide in the evaluation of current theories of the cortical mechanisms of working memory. PMID- 17418957 TI - Tumor necrosis factor-alpha induces changes in mitochondrial cellular distribution in motor neurons. AB - Motor neuron (MN) mitochondrial abnormalities and elevation in spinal fluid levels of the inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). The mechanism of neuron death in ALS remains unclear, along with the contributions of mitochondrial dysfunction and inflammation in the process. Cell cultures enriched for MN derived from embryonic rat spinal cords were established and directly exposed in vitro to recombinant TNF-alpha for varying lengths of time. Although cytokine exposure for up to 4 days failed to induce MN death, mitochondrial changes were observed shortly after initiating treatment. Our results demonstrate that TNF-alpha induced mitochondrial redistribution toward the soma in MN. We postulate that inflammation may precede, and in fact cause, the mitochondrial changes observed in ALS tissue. PMID- 17418958 TI - Dissociation of cholinergic function in spatial and procedural learning in rats. AB - The cholinergic system has long been known for its role in acquisition and retention of new information. Scopolamine, a muscarinic acetylcholine receptor antagonist impairs multiple memory systems, and this has promoted the notion that drug-induced side effects are responsible for diminished task execution rather than selective impairments on learning and memory per se. Here, we revisit this issue with the aim to dissociate the effects of scopolamine (0.2-1.0 mg/kg) on spatial learning in the water maze. Experiments 1 and 2 showed that acquisition of a reference memory paradigm with constant platform location is compromised by scopolamine independent of whether the animals are pre-trained or not. Deficits were paralleled by drug induced side-effects on sensorimotor parameters. Experiment 3 explored the role of muscarinic receptors in acquisition of an episodic-like spatial delayed matching to position (DMTP) protocol, and scopolamine still caused a learning deficit and side-effects on sensorimotor performance. Rats extensively pre-trained in the DMTP protocol with 30 s and 1 h delays over several months in experiment 4 and tested in a within-subject design under saline and scopolamine had no sensorimotor deficits, but spatial working memory remained compromised. Experiment 5 used the rising Atlantis platform in the DMTP paradigm. Intricate analysis of the amount of dwelling and its location revealed a clear deficit in spatial working memory induced by scopolamine, but there was no effect on sensorimotor or procedural task demands. Apart from the well-known contribution to sensorimotor and procedural learning, our findings provide compelling evidence for an important role of muscarinic acetylcholine receptor signaling in spatial episodic-like memory. PMID- 17418959 TI - Chordoma arising in a mature cystic teratoma of the ovary: a case report. AB - Mature cystic teratoma of the ovary (MCTO) is the most common type of ovarian teratoma and also the most frequent tumor originating from germ cells. It is usually diagnosed in early adulthood and, by definition, is composed of well differentiated tissues, which originate from all three germ cell layers. Unusual types of tissues can be found in MCTO, such as kidney, adrenal, and prostatic tissues. Malignant transformation is reported in less than 2% of teratomas. Squamous cell carcinoma is the most common malignancy arising in these otherwise benign tumors. We present the first case of MCTO containing a chordoma. The chordoma differentiation was supported by immunohistochemical staining and interphase fluorescence in situ hybridization (IP-FISH) technique showing 19% of the nuclei of the MCTO displaying polysomy for the chromosome X, while 28% of the chordoma nuclei showed chromosome 7 mosaicism. These results are concordant with previous studies, showing chromosomal anomalies in chromosomes X and 7 in MCTO and chordomas, respectively. PMID- 17418960 TI - Automatic delineation of on-line head-and-neck computed tomography images: toward on-line adaptive radiotherapy. AB - PURPOSE: To develop and validate a fully automatic region-of-interest (ROI) delineation method for on-line adaptive radiotherapy. METHODS AND MATERIALS: On line adaptive radiotherapy requires a robust and automatic image segmentation method to delineate ROIs in on-line volumetric images. We have implemented an atlas-based image segmentation method to automatically delineate ROIs of head-and neck helical computed tomography images. A total of 32 daily computed tomography images from 7 head-and-neck patients were delineated using this automatic image segmentation method. Manually drawn contours on the daily images were used as references in the evaluation of automatically delineated ROIs. Two methods were used in quantitative validation: (1) the dice similarity coefficient index, which indicates the overlapping ratio between the manually and automatically delineated ROIs; and (2) the distance transformation, which yields the distances between the manually and automatically delineated ROI surfaces. RESULTS: Automatic segmentation showed agreement with manual contouring. For most ROIs, the dice similarity coefficient indexes were approximately 0.8. Similarly, the distance transformation evaluation results showed that the distances between the manually and automatically delineated ROI surfaces were mostly within 3 mm. The distances between two surfaces had a mean of 1 mm and standard deviation of <2 mm in most ROIs. CONCLUSION: With atlas-based image segmentation, it is feasible to automatically delineate ROIs on the head-and-neck helical computed tomography images in on-line adaptive treatments. PMID- 17418961 TI - Analysis of prognostic factors and patterns of recurrence in patients with pathologic stage III endometrial cancer. AB - PURPOSE: To retrospectively assess prognostic factors and patterns of recurrence in patients with pathologic Stage III endometrial cancer. METHODS AND MATERIALS: Between 1989 and 2003, 107 patients with pathologic International Federation of Gynecology and Obstetrics Stage III endometrial adenocarcinoma confined to the pelvis were treated at our institution. Adjuvant radiotherapy (RT) was delivered to 68 patients (64%). The influence of multiple patient- and treatment-related factors on pelvic and distant control and overall survival (OS) was evaluated. RESULTS: Median follow-up for patients at risk was 41 months. Five-year actuarial OS was significantly improved in patients treated with adjuvant RT (68%) compared with those with resection alone (50%; p = 0.029). Age, histology, grade, uterine serosal invasion, adnexal involvement, number of extrauterine sites, and treatment with adjuvant RT predicted for improved survival in univariate analysis. Multivariate analysis revealed that grade, uterine serosal invasion, and treatment with adjuvant RT were independent predictors of survival. Five-year actuarial pelvic control was improved significantly with the delivery of adjuvant RT (74% vs. 49%; p = 0.011). Depth of myometrial invasion and treatment with adjuvant RT were independent predictors of pelvic control in multivariate analysis. CONCLUSIONS: Multiple prognostic factors predicting for the outcome of pathologic Stage III endometrial cancer patients were identified in this analysis. In particular, delivery of adjuvant RT seems to be a significant independent predictor for improved survival and pelvic control, suggesting that pelvic RT should be routinely considered in the management of these patients. PMID- 17418962 TI - Telomere shortening and associated chromosomal instability in peripheral blood lymphocytes of patients with Hodgkin's lymphoma prior to any treatment are predictive of second cancers. AB - PURPOSE: To investigate a potential link between telomere length, chromosomal instability, and the advent of a second cancer (SC) in patients with Hodgkin's lymphoma (HL), who are known to be at risk for SCs. This study was premised on the finding that telomere dysfunction and DNA repair pathways were related to many pathologic conditions. METHODS AND MATERIALS: Three cohorts of patients with HL were studied: 73 who were prospectively followed >5 years after diagnosis (prospective HL cohort), 28 who developed a SC (SC HL cohort), and 18 long-term survivors with no evidence of disease or complication since their initial treatment (NED HL cohort). Telomere length was analyzed by a telomeric restriction fragment assay in peripheral blood lymphocytes. Thirty healthy donors and 70 patients with a newly diagnosed solid tumor were the control population. RESULTS: Compared with controls, patients from the prospective HL cohort, before any treatment, showed age-independent shorter telomeres (mean, 8.3 vs. 11.7 kb in healthy donors; <6 kb in 18% in HL patients), increased spontaneous chromosomal abnormalities, and increased in vitro radiation sensitivity (p < 10(-4) each). After treatment, telomere shortening was associated with cytogenetic profiles characterized by the persistence of complex chromosomal rearrangement and clonal aberrations. Moreover, the two cases of SC in the prospective HL patients had short telomeres and CCR initially. In addition, the SC HL cohort was characterized by markedly short telomeres (6.6 vs. 9.7 kb in the NED HL cohort), the presence of complex chromosome rearrangements, and increased in vitro radiation sensitivity. CONCLUSIONS: An intimate relationship between pre treatment telomere shortening, chromosomal instability, radiation sensitivity and occurrence of SC was found in HL patients. PMID- 17418963 TI - Who enrolls onto clinical oncology trials? A radiation Patterns Of Care Study analysis. AB - PURPOSE: To identify factors significantly influencing accrual to clinical protocols by analyzing radiation Patterns of Care Study (PCS) surveys of 3,047 randomly selected radiotherapy (RT) patients. METHODS AND MATERIALS: Patterns of Care Study surveys from disease sites studied for the periods 1992-1994 and 1996 1999 (breast cancer, n = 1,080; prostate cancer, n = 1,149; esophageal cancer, n = 818) were analyzed. The PCS is a National Cancer Institute-funded national survey of randomly selected RT institutions in the United States. Patients with nonmetastatic disease who received RT as definitive or adjuvant therapy were randomly selected from eligible patients at each institution. To determine national estimates, individual patient records were weighted by the relative contribution of each institution and patients within each institution. Data regarding participation in clinical trials were recorded. The factors age, gender, race, type of insurance, and practice type of treating institution (academic or not) were studied by univariate and multivariate analyses. RESULTS: Overall, only 2.7% of all patients were accrued to clinical protocols. Of these, 57% were enrolled on institutional review board-approved institutional trials, and 43% on National Cancer Institute collaborative group studies. On multivariate analysis, patients treated at academic facilities (p = 0.0001) and white patients (vs. African Americans, p = 0.0002) were significantly more likely to participate in clinical oncology trials. Age, gender, type of cancer, and type of insurance were not predictive. CONCLUSIONS: Practice type and race significantly influence enrollment onto clinical oncology trials. This suggests that increased communication and education regarding protocols, particularly focusing on physicians in nonacademic settings and minority patients, will be essential to enhance accrual. PMID- 17418964 TI - Cone beam computed tomography guidance for setup of patients receiving accelerated partial breast irradiation. AB - PURPOSE: To evaluate the role of cone-beam CT (CBCT) guidance for setup error reduction and soft tissue visualization in accelerated partial breast irradiation (APBI). METHODS AND MATERIALS: Twenty patients were recruited for the delivery of radiotherapy to the postoperative cavity (3850 cGy in 10 fractions over 5 days) using an APBI technique. Cone-beam CT data sets were acquired after an initial skin-mark setup and before treatment delivery. These were registered online using the ipsilateral lung and external contours. Corrections were executed for translations exceeding 3 mm. The random and systematic errors associated with setup using skin-marks and setup using CBCT guidance were calculated and compared. RESULTS: A total of 315 CBCT data sets were analyzed. The systematic errors for the skin-mark setup were 2.7, 1.7, and 2.4 mm in the right-left, anterior-posterior, and superior-inferior directions, respectively. These were reduced to 0.8, 0.7, and 0.8 mm when CBCT guidance was used. The random errors were reduced from 2.4, 2.2, and 2.9 mm for skin-marks to 1.5, 1.5, and 1.6 mm for CBCT guidance in the right-left, anterior-posterior, and superior-inferior directions, respectively. CONCLUSION: A skin-mark setup for APBI patients is sufficient for current planning target volume margins for the population of patients studied here. Online CBCT guidance minimizes the occurrence of large random deviations, which may have a greater impact for the accelerated fractionation schedule used in APBI. It is also likely to permit a reduction in planning target volume margins and provide skin-line visualization and dosimetric evaluation of cardiac and lung volumes. PMID- 17418965 TI - Electromagnetic head-and-neck hyperthermia applicator: experimental phantom verification and FDTD model. AB - PURPOSE: To experimentally verify the feasibility of focused heating in the neck region by an array of two rings of six electromagnetic antennas. We also measured the dynamic specific absorption rate (SAR) steering possibilities of this setup and compared these SAR patterns to simulations. METHODS AND MATERIALS: Using a specially constructed laboratory prototype head-and-neck applicator, including a neck-mimicking cylindrical muscle phantom, we performed SAR measurements by electric field, Schottky-diode sheet measurements and, using the power-pulse technique, by fiberoptic thermometry and infrared thermography. Using phase steering, we also steered the SAR distribution in radial and axial directions. All measured distributions were compared with the predictions by a finite difference time-domain-based electromagnetic simulator. RESULTS: A central 50% iso-SAR focus of 35 +/- 3 mm in diameter and about 100 +/- 15 mm in length was obtained for all investigated settings. Furthermore, this SAR focus could be steered toward the desired location in the radial and axial directions with an accuracy of approximately 5 mm. The SAR distributions as measured by all three experimental methods were well predicted by the simulations. CONCLUSION: The results of our study have shown that focused heating in the neck is feasible and that this focus can be effectively steered in the radial and axial directions. For quality assurance measurements, we believe that the Schottky-diode sheet provides the best compromise among effort, speed, and accuracy, although a more specific and improved design is warranted. PMID- 17418966 TI - Postoperative irradiation of gynecologic malignancies: improving treatment delivery using aperture-based intensity-modulated radiotherapy. AB - PURPOSE: To evaluate dosimetric and treatment delivery advantages of aperture based intensity-modulated radiotherapy (AB-IMRT) for the treatment of patients receiving whole pelvic radiotherapy for gynecologic malignancies. METHODS AND MATERIALS: Nineteen patients undergoing pelvic radiotherapy after resection of endometrial cancers were selected. A 45-Gy dose was prescribed to the target volume delineated on a planning CT scan. An in-house inverse planning system, Ballista, was used to develop a treatment plan using aperture-based multileaf collimator segments. This approach was compared with conventional four-field, enlarged four-field, and static beamlet-based IMRT (BB-IMRT) techniques in terms of target coverage, dose-volume histogram statistics for surrounding normal tissues, and numbers of segments and monitor units (MU). RESULTS: Three quarters (76.4%) of the planning target volume received the prescription dose with conventional four-field plans. With adequate target coverage, the Ballista plans significantly reduced the volume of bowel and bladder irradiated at the prescribed dose (p < 0.001), whereas the two approaches provided equivalent results for the rectum (p = 0.5). On the other hand, AB-IMRT and BB-IMRT plans showed only small differences in dose-volume histogram statistics of unknown clinical impact, whereas Ballista plan delivery required on average 73% and 59% fewer segments and MU, respectively. CONCLUSION: With respect to conventional techniques, AB-IMRT for the treatment of gynecologic malignancies provides dosimetric advantages similar to those with BB-IMRT but with clear treatment delivery improvements. PMID- 17418967 TI - Comparison of outcomes for patients with unresectable, locally advanced non-small cell lung cancer treated with induction chemotherapy followed by concurrent chemoradiation vs. concurrent chemoradiation alone. AB - PURPOSE: To retrospectively compare outcomes for patients with unresectable locally advanced non-small-cell lung cancer (NSCLC) treated at our institution with concurrent chemoradiation with or without induction chemotherapy. METHODS AND MATERIALS: We retrospectively analyzed 265 consecutive patients who received definitive treatment with three-dimensional conformal radiation and concurrent chemotherapy. Of these, 127 patients received induction chemotherapy before concurrent chemoradiation. RESULTS: The two groups of patients (with induction vs. without induction chemotherapy) were similar in age, performance status, weight loss, histology, grade, and stage. Patients who received induction chemotherapy had better overall survival (median, 1.9 vs. 1.4 years; 5-year rate, 25% vs. 12%; p < 0.001) and distant metastasis-free survival (5-year rate, 42% vs. 23%; p = 0.021). Locoregional control was not significantly different between the two groups. Multivariate analysis showed that induction chemotherapy was the most significant factor affecting overall survival, with a hazard ratio of 0.55 (95% confidence interval 0.40-0.75; p < 0.001). A planned subgroup analysis showed that induction chemotherapy was associated with a significant overall survival benefit for patients with adenocarcinoma or large-cell carcinoma (5-year rate, 24% vs. 8%; p = 0.003) but not for those with squamous cell carcinoma. A multivariate analysis of patients with adenocarcinoma or large-cell carcinoma confirmed that induction chemotherapy was the most significant factor associated with better overall survival, with a hazard ratio of 0.47 (95% confidence interval, 0.28-0.78; p = 0.003). CONCLUSION: Our retrospective analysis suggests that in combination with concurrent chemoradiation, induction chemotherapy may provide a small but significant survival benefit for patients with unresectable locally advanced adenocarcinoma or large-cell carcinoma of the lung. PMID- 17418968 TI - Randomized phase II trial of high-dose melatonin and radiation therapy for RPA class 2 patients with brain metastases (RTOG 0119). AB - PURPOSE: To determine if high-dose melatonin for Radiation Therapy Oncology Group (RTOG) recursive partitioning analysis (RPA) Class 2 patients with brain metastases improved survival over historical controls, and to determine if the time of day melatonin was given affected its toxicity or efficacy. RTOG 0119 was a phase II randomized trial for this group of patients. METHODS AND MATERIALS: RTOG RPA Class 2 patients with brain metastases were randomized to 20 mg of melatonin, given either in the morning (8-9 AM) or in the evening (8-9 PM). All patients received radiation therapy (30 Gy in 10 fractions) in the afternoon. Melatonin was continued until neurologic deterioration or death. The primary endpoint was overall survival time. Neurologic deterioration, as reflected by the Mini-Mental Status Examination, was also measured. RESULTS: Neither of the randomized groups had survival distributions that differed significantly from the historic controls of patients treated with whole-brain radiotherapy. The median survivals of the morning and evening melatonin treatments were 3.4 and 2.8 months, while the RTOG historical control survival was 4.1 months. CONCLUSIONS: High-dose melatonin did not show any beneficial effect in this group of patients. PMID- 17418969 TI - Risk factors of ototoxicity after cisplatin-based chemo-irradiation in patients with locally advanced head-and-neck cancer: a multivariate analysis. AB - PURPOSE: Cisplatin chemo-irradiation is increasingly used in locally advanced squamous cell carcinoma of the head and neck. The objective of this study is to determine risk factors of ototoxicity due to intra-arterial high-dose cisplatin chemoradiation. METHODS AND MATERIALS: A prospective analysis of hearing thresholds at low and (ultra) high frequencies obtained before, during, and after treatment in 146 patients. Treatment consisted of intra-arterial infusion of high dose cisplatin (150 mg/m(2), four courses) with sodium thiosulfate rescue and concurrent radiation therapy (70 Gy). Patient and chemoradiation variables were studied in a multivariate analysis. RESULTS: After treatment, 23% of the ears were under consideration for hearing aids because of therapy. Twenty-two percent of the patients developed an increase in air-bone gap >10 dB during or after therapy. In the multivariate explanatory analysis, cumulative dose of cisplatin and radiation therapy, and young age displayed a causal relationship with increased sensorineural hearing loss during and after therapy (p < 0.001). In the multivariate prediction analysis, pretreatment hearing level of the concerning ear was identified as an independent predictive factor for hearing capability after therapy (p < 0.0001). CONCLUSIONS: Both cisplatin and radiation therapy were proven to induce sensorineural hearing loss, in this study with short-term follow-up. Of all patient and treatment variables studied, the patients pretreatment hearing level appeared to be the main predictive factor for hearing capability after high-dose intra-arterial cisplatin chemoradiation. PMID- 17418970 TI - A bystander effect observed in boron neutron capture therapy: a study of the induction of mutations in the HPRT locus. AB - PURPOSE: To investigate bystander mutagenic effects induced by alpha-particles during boron neutron capture therapy, we mixed cells that were electroporated with borocaptate sodium (BSH), which led to the accumulation of (10)B inside the cells, and cells that did not contain the boron compound. The BSH-containing cells were irradiated with alpha-particles produced by the 10B(n,alpha)7Li reaction, whereas cells without boron were affected only by the 1H(n,gamma)2H and 14N(n,rho)14C reactions. METHODS AND MATERIALS: The lethality and mutagenicity measured by the frequency of mutations induced in the hypoxanthine-guanine phosphoribosyltransferase locus were examined in Chinese hamster ovary cells irradiated with neutrons (Kyoto University Research Reactor: 5 MW). Neutron irradiation of 1:1 mixtures of cells with and without BSH resulted in a survival fraction of 0.1, and the cells that did not contain BSH made up 99.4% of the resulting cell population. The molecular structures of the mutations were determined using multiplex polymerase chain reactions. RESULTS: Because of the bystander effect, the frequency of mutations increased in the cells located nearby the BSH-containing cells compared with control cells. Molecular structural analysis indicated that most of the mutations induced by the bystander effect were point mutations and that the frequencies of total and partial deletions induced by the bystander effect were less than those induced by the original neutron irradiation. CONCLUSION: These results suggested that in boron neutron capture therapy, the mutations caused by the bystander effect and those caused by the original neutron irradiation are induced by different mechanisms. PMID- 17418971 TI - Radiation doses to structures within and adjacent to the larynx are correlated with long-term diet- and speech-related quality of life. AB - PURPOSE: To test the hypothesis that radiation dose to key sites in the upper aerodigestive tract is associated with long-term functional outcome after (chemo)radiotherapy for head-and-neck cancers. METHODS AND MATERIALS: This study examined the outcome for 27 patients treated with intensity-modulated radiotherapy for definitive management of their head-and-neck cancer who were disease free for at least 1 year after treatment. Head-and-neck cancer-specific quality of life (QoL) was assessed before treatment and at 1 year after treatment. Type of diet tolerated, presence of a feeding tube, and degree of weight loss 1 year after treatment were also used as outcome measures. Radiation doses delivered to various points along the upper aerodigestive tract, including base of tongue, lateral pharyngeal walls, and laryngeal structures, were determined from each treatment plan. Radiation doses for each of these points were tested for correlation with outcome measures. RESULTS: Higher doses delivered to the aryepiglottic folds, false vocal cords, and lateral pharyngeal walls near the false cords correlated with a more restrictive diet, and higher doses to the aryepiglottic folds correlated with greater weight loss (p < 0.05) 1 year after therapy. Better posttreatment speech QoL scores were associated with lower doses delivered to structures within and surrounding the larynx. CONCLUSION: Our data show an inverse relationship between radiation dose delivered to laryngeal structures and speech and diet and QoL outcomes after definitive (chemo)radiation treatment. These findings suggest that efforts to deliver lower doses to laryngeal structures may improve outcomes after definitive (chemo)radiation therapy. PMID- 17418972 TI - Predictive factors for late genitourinary and gastrointestinal toxicity in patients with prostate cancer treated with adjuvant or salvage radiotherapy. AB - PURPOSE: To determine the rate and magnitude of late genitourinary (GU) and gastrointestinal (GI) toxicities after salvage or adjuvant radiotherapy (RT) for prostate cancer, and to determine predictive factors for these toxicities. METHODS AND MATERIALS: A large multi-institutional database that included 959 men who received postoperative RT after radical prostatectomy (RP) was analyzed: 19% received adjuvant RT, 81% received salvage RT, 78% were treated to the prostate bed only, and 22% received radiation to the pelvis. RESULTS: The median follow-up time was 55 months. At 5 years, 10% of patients had Grade 2 late GU toxicity and 1% had Grade 3 late GU toxicity, while 4% of patients had Grade 2 late GI toxicity and 0.4% had Grade 3 late GI toxicity. Multivariate analysis demonstrated that adjuvant RT (p = 0.03), androgen deprivation (p < 0.0001), and prostate bed-only RT (p = 0.007) predicted for Grade 2 or higher late GU toxicity. For GI toxicity, although adjuvant RT was significant in the univariate analysis, no significant factors were found in the multivariate analysis. CONCLUSIONS: Overall, the number of high-grade toxicities for postoperative RT was low. Therefore, adjuvant and salvage RT can safely be used in the appropriate settings. PMID- 17418973 TI - Postmastectomy radiation improves the outcome of patients with locally advanced breast cancer who achieve a pathologic complete response to neoadjuvant chemotherapy. AB - PURPOSE: The aim of this study was to investigate the role of postmastectomy radiation therapy in women with breast cancer who achieved a pathologic complete response (pCR) to neoadjuvant chemotherapy. METHODS AND MATERIALS: We retrospectively identified 226 patients treated at our institution who achieved a pCR at surgery after receiving neoadjuvant chemotherapy. Of these, the 106 patients without inflammatory breast cancer who were treated with mastectomy were analyzed. The patients' clinical stages at diagnosis were I in 2%, II in 31%, IIIA in 30%, IIIB in 25%, and IIIC in 11% (American Joint Committee on Cancer 2003 system). Of the patients, 92% received anthracycline-based chemotherapy, and 38% also received a taxane. A total of 72 patients received postmastectomy radiation therapy, and 34 did not. The actuarial rates of local-regional recurrence (LRR) and survival of the two groups were compared using the log-rank test. RESULTS: The median follow-up of surviving patients was 62 months. Use of radiation therapy did not affect the 10-year rates of LRR for patients with Stage I or II disease (the 10-year LRR rates were 0% for both groups). However, the 10 year LRR rate for patients with Stage III disease was significantly improved with radiation therapy (7.3% +/- 3.5% with vs. 33.3% +/- 15.7% without; p = 0.040). Within this cohort, use of radiation therapy was also associated with improved disease-specific and overall survival. CONCLUSION: Postmastectomy radiation therapy provides a significant clinical benefit for breast cancer patients who present with clinical Stage III disease and achieve a pCR after neoadjuvant chemotherapy. PMID- 17418974 TI - Breast-conservative surgery with close or positive margins: can the breast be preserved with high-dose-rate brachytherapy boost? AB - PURPOSE: To evaluate the likelihood of preserving the breast in women who show close or positive margins after conservative surgery for early breast carcinoma. METHODS AND MATERIALS: Since 1996, 125 women with less than 5 mm or positive margins and positive separate cavity margin sampling were entered in a prospective trial with high-dose radiotherapy. A standard dose of 50 Gy to the whole breast was followed by a high-dose-rate brachytherapy application delivering 3 fractions of 4.4 Gy in 24 hours. The median follow-up was 84 months. RESULTS: There were only seven local recurrences, with an actuarial local control rate of 95.8% at 5 years and 91.1% at 9 years. Actuarial overall and cause specific survival rates were 92.6% and 95% at 5 years and 86.7% and 90.4% at 9 years, respectively. Late fibrosis was the most common complication, in 30% of patients, with good or excellent cosmetic results in 77%. The final result was that 95.2% of breasts were preserved. CONCLUSIONS: Close or positive-margin breast cancer can be well managed with a high-dose boost in a wide tumor bed by means of high-dose-rate brachytherapy. This technique can avoid mastectomy or poor cosmetic resection, with minimal risk of local or general failure. PMID- 17418975 TI - Stefin a and stefin B: markers for prognosis in operable squamous cell carcinoma of the head and neck. AB - PURPOSE: The aim of this study was to test the hypothesis about the protective role of high stefin A and stefin B concentrations in operable carcinoma of the head and neck. METHODS AND MATERIALS: Stefins A and B concentrations were measured in tissue cytosols of nontumorous mucosa and primary tumors from 92 patients. For quantitative analysis of stefins in tumor cytosols, commercially available enzyme-linked immunosorbent assays were used. RESULTS: Stefin A was upregulated in 53 patients (higher concentrations were measured in tumor samples than in nontumorous mucosa) and was downregulated in 39 patients. The corresponding numbers for stefin B were 49 and 43, respectively. A significantly higher proportion of downregulated cases were found among patients with disease re-appearance. In the Cox model, high stefin A concentrations appeared as independent predictors for favorable disease-free survival. Assuming a "broken stick" model, a significant increase in the recurrence rate after the threshold of 1063 ng/mgp (the 64th percentile in the group) was found, the hazard ratio reaching 3% of the reference value with doubling of the level of stefin A. These results were reconfirmed after pooling the data with two historical data sets into a uniform series involving 182 patients. CONCLUSIONS: A group of patients at high risk for disease progression was identified, characterized by the downregulated stefin A protein in the tumor compared with the nontumorous mucosa. Stefin A was recognized as a promising candidate marker for prognosis in patients with operable carcinoma of the head and neck. PMID- 17418976 TI - Nutritional and lifestyle correlates of the cancer-protective hormone melatonin. AB - CONTEXT: Despite growing support for melatonin as a promising agent for cancer treatment and possibly cancer prevention, few studies have elucidated factors that influence endogenous melatonin. This overview summarizes dietary and lifestyle factors that have been shown to affect circulating melatonin levels. BIOLOGICAL MECHANISMS: To date, many animal studies and in vitro experiments have illustrated that melatonin possesses oncostatic activity. Mechanisms that are currently being studied include melatonin's activity as an indirect antioxidant and free radical scavenger; its action on the immune system; suppression of fatty acid uptake and metabolism; and its ability to increase the degradation of calmoduline and to induce apoptosis. Studies further suggest that melatonin reduces local estrogen synthesis, through down-regulation of the hypothalamic pituitary reproductive axis and direct actions of melatonin at the tumor cell level, thus behaving as a SERM. THERAPEUTIC APPLICATIONS: Several small clinical trials have demonstrated that melatonin has some potential, either alone or in combination with standard cancer therapy, to yield favorable responses. Melatonin or its precursor tryptophan have been found in numerous edible plants, but more studies are needed to evaluate the influence of diets rich in tryptophan and melatonin on circulating melatonin levels in humans. Age, BMI, parity, and the use of certain drugs remain the factors that have been associated most consistently with aMT6s levels. DISCUSSION: Further insights into the effects of dietary and lifestyle factors that modulate circulating melatonin levels may provide the basis for novel interventions to exploit melatonin for the prevention and treatment of human diseases. PMID- 17418977 TI - Pretreatment total testosterone levels in patients with prostate cancer in the past two decades in Japan. AB - BACKGROUND: Many studies have shown the relationship between pretreatment serum testosterone levels and the clinical stage or histological grade, but the clinical significance of pretreatment testosterone levels is controversial. We studied the association of pretreatment total testosterone levels with the clinical stage and histological grade of prostate cancer. METHODS: We evaluated 2914 patients whose pretreatment testosterone levels were recorded from 1982 to 2002. Serum testosterone levels were measured by radioimmunoassay. RESULTS: There was a trend toward decreasing testosterone values with worsening clinical staging. There was a trend toward decreasing testosterone values with worsening histological grading, too. Patients with poorly differentiated adenocarcinoma had significantly lower testosterone levels than those with the others (versus well; p<0.01, moderately; p<0.01). CONCLUSIONS: Newly diagnosed patients with poorly differentiated adenocarcinoma of prostate have lower testosterone levels than the others. PMID- 17418978 TI - "Health is strength": a community health education program to improve breast and cervical cancer screening among Korean American Women in Alameda County, California. AB - BACKGROUND: A 48-month community intervention was conducted to improve breast and cervical cancer (BCC) screening among Korean American (KA) women in Alameda County (AL), California. KA women in Santa Clara (SC) County, California served as a comparison group. METHODS: Random samples of KA women from each county were surveyed by telephone in 1994 (n=818) and 2002 (n=1084). Propensity score analyses were used to estimate the difference between counties in changes over time in screening (Pap tests, breast self-examinations, clinical breast examinations, and mammography), and to estimate differences in screening between participants and non-participants in an educational workshop among women in AL in 2002. RESULTS: Mammography screening and clinical breast examinations increased over time in both counties. Pap tests increased in AL but not SC, and breast self examinations did not change significantly in either county. None of the intervention-comparison group differences over time were significant. In 2002, compared to non-participants, women who attended a workshop were more likely to report a recent Pap test (P<.08). CONCLUSIONS: Although our overall intervention did not appear to enhance screening practices at the community-level, attendance at a women's health workshop appears to have increased cervical cancer screening. PMID- 17418979 TI - An investigation of relationships between hypoxia-inducible factor-1 alpha gene polymorphisms and ovarian, cervical and endometrial cancers. AB - BACKGROUND: DNA sequence variations in HIF-1 alpha gene might yield changes both in the production outcomes and in the activities of the gene. Overexpression of the HIF-1 alpha subunit, resulting from intratumoral hypoxia and genetic alterations, has been demonstrated in common human cancers and is correlated with tumor angiogenesis and patient mortality. In this study, we aimed to determine how the three single nucleotide polymorphisms (SNPs, C1772T and G1790A exon 12, C111A exon 2) in the HIF-1 alpha gene coding regions affect the ovarian, cervical and endometrial cancer patients in the Turkish population. A study on this relationship has not been conducted to date. METHOD: 102 gynecologic cancer patients and 107 healthy controls were studied. Genotypes of the three polymorphisms were analyzed by PCR-RFLP. RESULTS: There was no significant difference between ovarian cancer patients and controls in terms of the distribution of C1772T genotypes and alleles (P>0.05). However, there was a highly significant increase in the frequency of both CT 1772 and TT 1772 genotypes in patients with cervical and endometrial cancers compared with healthy controls. In fact, 1772T allele-carriers (CT+TT genotypes) showed an association with the risk of cervical and endometrial cancers compared to the wild type (OR=3.84, 95% CI: 1.65-8.93; OR=7.41, 95% CI: 2.33-23.59, respectively). C1772T polymorphism was not associated with family history concerning gynecologic and/or other cancer types, stages (I-IV) and grades of tumor, smoking habits and existence of other diseases that generate a hypoxic microenvironment even after multivariable logistic regression analysis. As for HIF-1 alpha G1790A genotypes, the frequencies of G alleles were 98% in ovarian patients and 100% in the control group. We found no significant difference in the genotype distribution and allele frequencies between the ovarian patients and healthy control subjects. There were no GA and AA genotypes among the cervical and endometrial cancer patients. As for HIF-1 alpha C111A polymorphism, we did not find CA and AA variants of the gene in controls or in any of the three types of patients. CONCLUSION: Our results suggest that the C1772T polymorphism of the HIF-1 alpha may be associated with cervical and endometrial cancers. PMID- 17418980 TI - Mammography screening in six diverse communities in Chicago--a population study. AB - BACKGROUND: Despite the fact that recent studies suggest a narrowing in access to mammography, Black women are much more likely to die from breast cancer than White women. Data at the community level regarding mammography screening can help explain health disparities and inform plans for improved screening efforts. METHODS: In 2002-2003, a comprehensive household health survey in English or Spanish was conducted in six community areas with 1700 households. The module on mammography was based on a state-based nationwide health survey and included questions on frequency of mammography, repeat screenings, and several demographic variables. RESULTS: The proportion of women >or=40 years (n=482) who received a mammogram in the past 2 years ranged from 74% to 90% across the six communities. The community with the highest screening proportion was predominantly Mexican and included recent immigrants. The screening proportion in the poorest community area, which was all Black, was 77%. Women with health insurance, higher income, and more education were more likely to receive a mammogram. Proportions for women >or=50 years (n=286) were slightly higher but similar. Repeat screening, which is recommended, occurred at lower levels. CONCLUSIONS: Access to and utilization of mammography have grown in recent years so that even these vulnerable communities had screening proportions at or even higher than the national average and the Healthy People Year 2010 objective. Nonetheless, repeat screening sequences were lower and may require attention if mammography screening efforts are to have a greater impact on female breast cancer mortality. PMID- 17418981 TI - Willow bark extract (BNO1455) and its fractions suppress growth and induce apoptosis in human colon and lung cancer cells. AB - BACKGROUND: Recently, there have been extensive efforts to evaluate the chemopreventive role of substances present in natural products. The aim of this study was to examine the effects of the main groups of compounds (salicylalcohol derivates, flavonoids, proanthocyanidins), and salicin isolated from willow bark extract BNO 1455 on proliferation and apoptosis in human colon and cancer cells. METHODS: We used human colon cyclooxygenase-2 (COX-2)-positive HT 29 and (COX-2) negative HCT 116 or lung COX-2 proficient A 549 and low COX-2 expressing SW2 cells. After treatment for 72 h with various concentrations of single substances and acetylsalicylic acid (ASA) as control, inhibition of cell growth and cytotoxicity were measured by colorimetric WST-1 assay and propidium iodide uptake by flow cytometry, respectively. Apoptotic cells were identified by annexin V adhesion using flow cytometry. RESULTS: Studies on dose-dependent effects of BNO 1455 and its fractions showed anti-proliferative activity of all compounds with 50% maximal growth inhibitory concentrations (GI(50)) between 33.3 and 103.3 microg/ml for flavonoids and proanthocyanidins fractions and 50.0-243.0 microg/ml for salicylalcohol derivates and extract. Apoptosis induction was confirmed by annexin V adherence and analysis of cell morphology based on light scattering characteristics using flow cytometry in all cell lines at GI(50). CONCLUSIONS: We showed that willow bark extract BNO 1455 an its fractions inhibit the cell growth and promote apoptosis in human colon and lung cancer cell lines irrespective of their COX-selectivity. PMID- 17418983 TI - [Maternal and foetal prognostic in excised women delivery]. AB - INTRODUCTION: The female circumcision constitutes by their frequency and complications a real problem of public health. MATERIAL AND METHOD: Our study aims at comparing the maternal land fetal complications of the spontaneous vaginal delivery in the excised women and non-excised. We led a comparative survey case witness implying 227 excised pregnant women at the maternity in CHU YO of Ouagadougou. RESULTS: The prevalence of the excision from January 1st to July 31, 2006 was 72.86%. The distribution of female genital mutilations in this population is the following: type I=27.75%, type II=69.61%, type III=2.64%. The middle age was 25 years and 79.30% of women were aged less than 30 years. Islam appeared like a factor of exposure to the practice of the excision with 67.40% of women excised that practise it against 41.90% at the non-excised group (P<0,0001). The maternal complications were dominated by the duration of fetal expulsion prolonged and perineal tears. The duration of fetal expulsion was superior to 30 minutes for 34.56% of excised woman childbirths 9 times more frequently than women non-excised (P=0.001). The frequency of perineal tears was 10.13% in the group of women excised against 5.73% in the group of the non excised (P=0.008). These perineal lesions were more frequent with the primiparae and women excised at the 2nd and 3rd degree. The neobirth asphyxia affected 4.4% of newborns from mother excised against 0.2% in the non-excised group (RR=5.18; P=0.006). In the group of excised them the rate of mortinatality was 22.03 for 1000 births, against 8.81 for 1000 births in the group of the non-excised (P=0.22). CONCLUSION: The prevention of these complications with the excised woman rests on the episiotomy and the instrumental extraction in the FGM of type III. PMID- 17418982 TI - Inhibition of cell proliferation, induction of apoptosis, reactivation of DLC1, and modulation of other gene expression by dietary flavone in breast cancer cell lines. AB - BACKGROUND: Dietary flavone was previously shown to increase the expression of deleted in liver cancer-1 gene (DLC-1) in HT-29 colon carcinoma cell line [Herzog A, Kindermann B, Doring F, Daniel H, Wenzel U. Pleiotropic molecular effects of the pro-apoptotic dietary constituent flavone in human colon cancer cells identified by protein and mRNA expression profiling. Proteomics 2004;4:2455-64]. DLC-1 that encodes a Rho GTPase-activating protein, functions as a tumor suppressor gene and is frequently inactivated or down-regulated in several common cancers. Restoration of DLC-1 expression suppresses in vitro tumor cells proliferation and tumorigenicity in vivo. METHODS: Here, the effect of flavone was examined in several DLC-1-deficient cell lines derived from different types human cancer using assays for cell proliferation, gene expression and transfer. RESULTS: We show that exposure to 150 microM flavone increased DLC1 expression in breast but not in liver or prostate carcinoma cells or a nonmalignant breast epithelial cell line. Flavone restored the expression of DLC1 in the breast carcinoma cell lines MDA-MB-468, MDA-MB-361, and BT20 as well as in the colon carcinoma cell line HT-29 all of which are DLC-1-negative due to promoter hypermethylation. We further show that flavone inhibited cell proliferation, induced cell cycle arrest at G(2)-M, increased p21(Waf1) gene expression, and caused apoptosis. Microarray analysis of these aggressive and metastatic breast carcinoma cells revealed 29 flavone-responsive genes, among which the DNA damage inducible GADD genes were up-regulated and the proto-oncogene STMN1 and IGFBP3 were down-regulated. CONCLUSIONS: Flavone-mediated alterations of genes that regulate tumor cell proliferation, cell cycle, and apoptosis contribute to chemopreventive and antitumoral effects of flavone. Alone or in combination with demethylating agents, flavone may be an effective adjunct to chemotherapy in preventing breast cancer metastasis. PMID- 17418984 TI - Quantitative behavior analysis and human values. AB - Many scientists believe that among the virtues of quantitative science are that its facts are free from personal, social, political, economic, and other cultural influences, or at least, if they are not, they should be. Radical behaviorism suggests, however, that a science of behavior must apply to peoples' everyday professional behaviors, including those of quantitative behavior analysts. The behaviors of quantitative behavior analysts, however, like the behaviors of everyone else, depend on the cultures to which they belong. A quantitative science of behavior must therefore describe and explain the cultural and human values of quantitative behavior analysts. In this sense, a quantitative science of behavior must apply to itself. No such "reflexive behavior analysis" currently exists and its development might shed considerable light on the basic nature of behavior analysis. PMID- 17418985 TI - Streptococcus iniae: an aquatic pathogen of global veterinary significance and a challenging candidate for reliable vaccination. AB - Streptococcus iniae has become one the most serious aquatic pathogens in the last decade causing high losses in farmed marine and freshwater finfish in warmer regions. Although first identified in 1976 from a captive Amazon freshwater dolphin, from which it derives its name, disease outbreaks had most likely been occurring for several decades in marine aquaculture in Japan. S. iniae is globally distributed throughout warm water finfish aquaculture. In common with other encapsulated beta-haemolytic streptococci and in direct contradiction to the phenomenal success story of bacterial vaccines in finfish aquaculture, control of S. iniae by vaccination has met with limited success. Thus, antibiotic usage is the current practice for reducing mortality and consequent economic loss. Vaccine failure appears to result in part from serotypic variation and, whilst 2 serotypes have been named, variation would appear to be more complex. S. iniae also has zoonotic potential, with human infections identified in the USA, Canada, and throughout Asia. In humans, infection is clearly opportunistic with all cases to date associated with direct infection of puncture wounds during preparation of contaminated fish, and generally in elderly or immunocompromised individuals. Significant progress has been made in terms of research into pathogenic mechanisms of S. iniae, with recent research elucidating the role of capsule in virulence for fish through antiopsonic activity. In light of this recent coverage in the literature, the present review centres on areas of direct veterinary interest including identification, epidemiology, therapy and prevention in farmed finfish. Clearly as the prevalence of S. iniae and associated economic losses continue to increase, further work towards developing a reliable vaccine is essential. This would appear to require a much better understanding of cell-surface variability amongst S. iniae isolates. PMID- 17418986 TI - Transport of stavudine, delavirdine, and saquinavir across the blood-brain barrier by polybutylcyanoacrylate, methylmethacrylate-sulfopropylmethacrylate, and solid lipid nanoparticles. AB - Permeability of the anti-human immunodeficiency virus (HIV) agents, including stavudine (D4T), delavirdine (DLV), and saquinavir (SQV), across the in vitro blood-brain barrier (BBB) was studied. Here, the anti-HIV agents were incorporated with polybutylcyanoacrylate (PBCA) nanoparticles (NPs), methylmethacrylate-sulfopropylmethacrylate (MMA-SPM) NPs, and solid lipid nanoparticles (SLNs). Transport of the anti-HIV agents across BBB is a key factor in their applications to the therapy of the acquired immunodeficiency syndrome (AIDS). Experimental results revealed that the drug order of the loading efficiency (LE) on PBCA and MMA-SPM was D4T>DLV>SQV. For the entrapment efficiency (EE) in SLNs, this order was reversed. Also, LE of D4T on MMA-SPM was larger than that on PBCA; however, the reverse was true for DLV and SQV. As the particle size increased, LE decreased and EE increased. For a fixed drug carrier, an increase in the particle size yielded a decrease in the BBB permeability coefficient of the anti-HIV agents. Moreover, enhancement in the BBB permeability was on the carrier order of PBCA>MMA-SPM>SLNs for D4T, and for DLV and SQV, the order became PBCA>SLNs>MMA-SPM. PBCA, MMA-SPM, and SLNs were efficacious carriers of D4T, DLV, and SQV to meliorate BBB permeability by 3-16 folds, indicating the clinical potential of the present NP formulations for the AIDS treatment. PMID- 17418987 TI - Ethnobotanical study of medicinal plants around 'Dheeraa' town, Arsi Zone, Ethiopia. AB - A study aimed at identifying plant species used and manipulated by the community for medicine was carried out around 'Dheeraa' town, Arsi Zone, Oromia Region, Southeast Ethiopia. The data were collected through a series of fieldworks conducted from October to December 2002. Random and systematic sampling methods were employed to select the study sites as well as the informants. Ethnobotanical methods using semi-structured interviews were employed and a total of 83 species of medicinal plants were recorded. The natural environment yielded larger proportion (76 species, 92%) of the total records followed by home gardens of the area. The studied medicinal plants were those used in traditional health care of humans and livestock. However, only very few individuals of each species were found in the area and this might be attributed to the ongoing habitat modification and loss of natural vegetation. Therefore, it is important to create awareness on sustainable use of the natural vegetation. Paying special attention to the medicinal plants found in the area may help to amplify the role that these plants play in healthcare, poverty alleviation as well as environmental protection. PMID- 17418988 TI - Evaluation of the anti-ulcerogenic effect of sesquiterpene lactones from Centaurea solstitialis L. ssp. solstitialis by using various in vivo and biochemical techniques. AB - The guaianolide type sesquiterpene lactones chlorojanerin, 13-acetyl solstitialin A and solstitialin A were identified as the anti-ulcerogenic components of the chloroform extract of the aerial parts of Centaurea solstitialis ssp. solstitialis (Asteraceae). In this study, these compounds were investigated by using various in vivo ulcer models in rats and mice. Chlorojanerin was shown to be significantly effective in preventing the induction of lesions by ethanol- (EtOH-) (both oral and subcutaneous administration), indomethacin-, indomethacin plus HCl/EtOH-, N(G)-nitro-l-arginine methyl ester plus EtOH-, N-ethylmaleimide plus EtOH-, water immersion and restraint stress, and serotonin, as well as inhibiting titratable gastric acidity and acid output, and increasing gastric pH, but was ineffective in the prevention of ulcers induced by pyloric ligation, diethyldithiocarbamate, and cysteamine, and had no effect on gastric secretion volume or peptic activity. A mixture of 13-acetyl solstitialin A (95%) and solstitialin A (5%) was found to be significantly effective against EtOH-induced lesions on oral administration but was ineffective when administered subcutaneously. This mixture was also found to be effective in preventing lesions induced by EtOH, indomethacin, indomethacin plus HCl/EtOH, N(G)-nitro-l-arginine methyl ester plus EtOH, N-ethylmaleimide plus EtOH, water immersion and restraint stress, serotonin and cysteamine, as well as inhibiting titratable gastric acidity and titratable acid output, and gastric pH, but was found ineffective against the pyloric ligation-induced and diethyldithiocarbamate-induced ulcerogenesis models, as well as gastric secretion volume and peptic activity. On the other hand, active compounds did not show any toxic effect on acute toxicity (3 days administration) evaluation tests in mice. PMID- 17418989 TI - Classification and individualization of black ballpoint pen inks using principal component analysis of UV-vis absorption spectra. AB - The technique of principal component analysis has been applied to the UV-vis spectra of inks obtained from a wide range of black ballpoint pens available in the UK market. Both the pen ink and material extracted from the ink line on paper have been examined. Here, principal component analysis characterised each spectrum within a group through the numerical loadings attached to the first few principal components. Analysis of the spectra from multiple measurements on the same brand of pen showed excellent reproducibility and clear discrimination between inks that was supported by statistical analysis. Indeed it was possible to discriminate between the pen ink and the ink line from all brands examined in this way, suggesting that the solvent extraction process may have an influence on these results. For the complete set of 25 pens, interpretation of the loadings for the first few principal components showed that both the pen inks and the extracted ink lines may be classified in an objective manner and in agreement with the results of parallel thin layer chromatography studies. Within each class almost all inks could be individualised. Further work has shown that principal component analysis may be used to identify a particular ink from a database of reference UV-vis spectra and a strategy for developing this approach is suggested. PMID- 17418990 TI - Prevalence of psychoactive substances in truck drivers in the Nord-Pas-de-Calais region (France). AB - A previous study conducted in 1995 showed that psychoactive drug use by workers was higher in safety/security workstations than in the rest of the labour force. In order to verify this finding, we conducted a new study in 2003-2004 in the Nord-Pas-de-Calais region, restricted to truck drivers. The aim of this study was to allow harmonizing the professional practice of the occupational physicians, proposing drug prevention and drug testing policies, validating the analytical methods and the guidelines in case of positive testing results. One thousand truck drivers were studied. Urines were tested for amphetamines, cannabinoids, cocaine, opiates, benzodiazepines, buprenorphine and methadone by immunoassay. Urine ethanol determinations were performed by an ADH method. Positive urines for drugs of abuse, methadone or buprenorphine were then tested by gas chromatography or liquid chromatography coupled to mass spectrometry. Out of the 1000 drivers, cannabinoids were detected in 85 cases, opiates in 41 cases, amphetamines in 3 cases and cocaine in only one case. Buprenorphine was detected in 18 cases, methadone in 5 cases and benzodiazepines in 4 cases. Urine ethanol was positive in 50 cases. We found only one case with 6-monoacetylmorphine. Other positive opiates were metabolites of antitussives. The relatively low number of benzodiazepine positive urines could be explained by the lack of sensitivity of the test we used. All these results confirm those of the previous study for cannabinoids and ethanol in safety/security workstations. Positive results for methadone and buprenorphine are eight times higher than in the general population. In conclusion, the authors think that it will be of a great interest to test urine of truck drivers for other classes of psychoactive drugs, using a liquid chromatography-mass spectrometry method. PMID- 17418991 TI - Muscle contractile properties in children with spastic diplegia. AB - The aim of the present study was to evaluate contractile properties of the plantarflexor muscles in children with spastic diplegia (SD) in comparison of age matched healthy children. Twelve prepubertal children with SD aged 11-12 years (6 girls and 6 boys) and 12 age- and gender-matched healthy control children (6 girls and 6 boys) participated in this study. Subjects were seated in a custom made dynamometric chair with the dominant leg flexed 90 degrees at the knee and ankle joints. Twitch contraction characteristics of the plantarflexor muscles were measured by supramaximal electrical stimulation of posterior tibial nerve in popliteal fossa using square-wave pulses of 1 ms duration at rest and after a brief (5 s) isometric maximal voluntary contraction (MVC), i.e., during post activation potentiation (PAP). Children with SD had significantly lower (p<0.05) MVC force, twitch contraction peak force (PF), PAP of twitch force, and twitch maximal rates of force development and relaxation compared to control group. Twitch contraction PF:MVC force ratio was higher (p<0.05) in children with SD than in the control group. However, no significant differences in twitch contraction and half-relaxation times were observed between the measured groups. It was concluded that prepubertal children with SD in comparison of normal children are characterized by markedly reduced isometric voluntary and electrically evoked twitch contraction maximal force, capacity for twitch PAP, and rates of twitch force production and relaxation of the plantarflexor muscles. The time-course characteristics of isometric twitch contraction were similar in children with SD and normal children. PMID- 17418992 TI - Impairment of methionine sulfoxide reductase during UV irradiation and photoaging. AB - During chronic UV irradiation, which is part of the skin aging process, proteins are damaged by reactive oxygen species resulting in the accumulation of oxidatively modified protein. UV irradiation generates irreversible oxidation of the side chains of certain amino acids resulting in the formation of carbonyl groups on proteins. Nevertheless, certain amino acid oxidation products such as methionine sulfoxide can be reversed back to their reduced form within proteins by specific repair enzymes, the methionine sulfoxide reductases A and B. Using quantitative confocal microscopy, the amount of methionine sulfoxide reductase A was found significantly lower in sun-exposed skin as compared to sun-protected skin. Due to the importance of the methionine sulfoxide reductase system in the maintenance of protein structure and function during aging and conditions of oxidative stress, the fate of this system was investigated after UVA irradiation of human normal keratinocytes. When keratinocytes are exposed to 15 J/cm(2) UVA, methionine sulfoxide reductase activity and content are decreased, indicating that the methionine sulfoxide reductase system is a sensitive target for UV induced inactivation. PMID- 17418993 TI - Development and full validation of six inhibition assays for five major cytochrome P450 enzymes in human liver microsomes using an automated 96-well microplate incubation format and LC-MS/MS analysis. AB - Substrate inhibition assays for five of the major CYP enzymes (phenacetin for CYP1A2, diclofenac for CYP2C9, (S)-mephenytoin for CYP2C19, dextromethorphan for CYP2D6 and midazolam and testosterone for CYP3A4) in human liver microsomes were developed. Fully automated incubations were conducted in a 96-well format under optimized enzyme kinetic conditions. Metabolites of probe substrates were analyzed with rapid LC-MS/MS methods. The assays were fully validated following the procedure for validating bioanalytical methods recommended by regulatory agencies. Quality control samples and a positive control CYP inhibitor were included in each assay. The IC(50) values determined for typical CYP inhibitors were reproducible and consistent with those reported in the literature. The high quality and throughput of these assays make them ideally suited for providing information for decision making in late drug discovery and early development and for providing labeling input for new drug registrations. PMID- 17418994 TI - Morbidity and mortality of laparoscopic vs. open radiofrequency ablation for hepatic malignancies. AB - AIMS: Surgical radiofrequency ablation (RFA) of hepatic malignancies is associated with superior oncological outcome as compared to percutaneous RFA. The aim of this prospective non-randomized cohort study was to compare morbidity and mortality of laparoscopic (LRFA) vs. open (ORFA) radiofrequency ablation of liver cancer. METHODS: Between October 1999 and November 2006, RFA was performed in 154 consecutive patients (percutaneous 12, LRFA 93, ORFA 49) for a total of 291 hepatic tumours (HCC 81, colorectal metastases 157, other 53). Seventy-four patients simultaneously underwent additional surgery. Laparoscopic RFA was performed in 45/54 patients with HCC, and in 44/54 patients with cirrhosis. Laparotomy was performed in 14/22 patients who underwent simultaneous colorectal resection, and in 12/22 patients with hepatic resection. RESULTS: Postoperative complications occurred in 25 patients with subsequent mortality in 2. As compared with LRFA, ORFA was associated with significantly (p<0.01) higher intra-operative blood loss (median 20 (range 0-1700) vs. 10 (0-900) ml), longer duration of surgery (180 (25-440) vs. 75 (30-390) min), more postoperative complications (17 vs. 8), and longer postoperative hospital stay (8 (1-127) vs. 4 (1-51) d). According to the therapy-oriented severity grading system (TOSGS) classification, postoperative complications in the ORFA-group were more severe than those in the LRFA-group (p<0.01). These findings were consistent in patients without simultaneous colorectal and/or hepatic resection and in patients with liver tumours measuring 3cm or less. In univariate analysis the following factors were significantly (p<0.01) related to the presence of postoperative complications: simultaneous colorectal resection, laparotomy, duration of surgery, tumour location in right liver, liver segment 7 (p=0.01), absence of cirrhosis (p=0.02), liver segment 8 (p=0.03), and metastatic liver cancer (p=0.04). CONCLUSION: LRFA for hepatic malignancies seems preferable above ORFA, provided good patient selection, surgical expertise, and long-term oncological control. PMID- 17418995 TI - The significance of the aberrant left hepatic artery arising from the left gastric artery at curative gastrectomy for gastric cancer. AB - BACKGROUND: An aberrant left hepatic artery (ALHA) is occasionally encountered during esophagogastric surgery. However, at curative gastrectomy for gastric cancer, it is questionable as to whether the ALHA need to be divided in order to maximize lymph node clearance and the issue requires clarification. METHODS: We encountered 50 patients with an ALHA during curative gastrectomy for gastric cancer between 1997 and 2001. Data concerning operative feasibility, postoperative liver function and therapeutic value of nodal dissection were analyzed retrospectively. RESULTS: For 27 patients, we preserved the ALHA, and for the remaining 23 patients, we divided the ALHA at the origin of the left gastric artery (LGA). Serum levels of aspartate aminotransferase and alanine aminotransferase were statistically significant higher on postoperative day (POD) 1 (P=0.0008 and P=0.0007), and on POD 3 (P=0.001 and P=0.008), respectively, in the ALHA-divided group. Patients who underwent a total gastrectomy predominated in the ALHA-divided group, the total number of dissected lymph nodes being higher in the ALHA-divided group (P=0.018). However, the total numbers of dissected lymph nodes and metastatic lymph nodes around the LGA were similar in the 2 groups (P=0.447 and P=0.128), respectively. No significant differences were seen between the 2 groups in morbidity and mortality. The overall 5-year survival rates were also comparable. CONCLUSIONS: Although a prospective study is required, this study suggested that routine division of the ALHA may not always be required for curative gastrectomy. PMID- 17418996 TI - [A case of spontaneous intracranial hypotension treated with lumbar blood patch]. PMID- 17418997 TI - [Evaluation of the renal function in cardiac surgery with CPB: role of the cystatin C and the calculated creatinine clearance]. AB - OBJECTIVES: The evaluation of the renal function in cardiac surgery is difficult. The gold standard remains the creatinine clearance in clinical practice. Cystatin C was recently proposed in order to evaluate the renal function. The aim of our study was to evaluate the cystatin C in cardiac surgery with CPB. PATIENTS AND METHODS: After informed consent and ethical committee agreement, 60 patients operated in cardiac surgery with CPB were prospectively included. Cystatin C,measured and calculated (Cockcroft and MDRD methods) creatinine were compared with the Student t-test and with the Bland and Altman method. p<0,05 was considered as a significant threshold. RESULTS: The reproducibility of the calculated creatinine clearance was better when the urinary collecting time was below 400 minutes. The estimation of the creatinine clearance by the Cockcroft and MDRD methods is better when the clearance is low. A significant correlation between the creatinine clearance and the cystatin C does exist, but the correlation coefficient was low. In case of acute renal dysfunction, the increase of the creatinine occurred earlier than the increase of the cystatin C. CONCLUSION: In cardiac surgery with CPB, the evaluation of the renal function was not improved by the cystatin C. PMID- 17418998 TI - Effect of berberine on hepatocyte proliferation, inducible nitric oxide synthase expression, cytochrome P450 2E1 and 1A2 activities in diethylnitrosamine- and phenobarbital-treated rats. AB - This study investigated the effect of berberine on the early phase of hepatocarcinogenesis stimulated by diethylnitrosamine (DEN, 150 mg/kg, 4 weeks) plus phenobarbital (PB, 75 mg/kg, 7 days) in rats. The expressions of proliferating cell nuclear antigen (PCNA) and inducible nitric oxide synthase (iNOS) were evaluated by immunohistochemistry. The activities of CYP isoenzymes were analyzed using different probe drugs including chlorzoxazone (CYP2E1) and phenacetin (CYP1A2) by high-performance liquid chromatography (HPLC) in vivo or in vitro. Results showed that the expressions of PCNA and iNOS were induced by DEN plus PB in liver tissues. Oral administration of berberine (50mg/kg) inhibited the hepatocyte proliferation and iNOS expression, decreased cytochrome P450 content, inhibited activities of CYP2E1 and CYP1A2 in DEN-plus-PB-treated rats in vivo. Moreover, berberine (10, 50 and 100 microM) inhibited the activities of CYP2E1 and CYP1A2 in microsomes isolated from DEN-plus-PB-treated rats in vitro, suggesting that anti-hepatocarcinogenetic potential of berberine might be due to inhibiting oxidative metabolic activities of CYP 2E1 and CYP1A2, and decreasing NO production in rats. PMID- 17418999 TI - Synthesis and anti-inflammatory effect of lipoxins in human airway epithelial cells. AB - In this study, we investigated the synthesis of lipoxins (LXs) and their anti inflammatory effects in different human airway epithelial cell culture models. After cell incubation with exogenous 5(S),6(R)-dihydroxy-7,9-trans-11,14-cis eicosatetraenoic acid, LXA(4) was detected in supernatants of differentiated human bronchial epithelial cells by contrast to non-differentiated cells. Exogenous LXA(4) significantly inhibited tumor necrosis factor-alpha (TNF-alpha) induced interleukin-8 (IL-8) release in the different epithelial cell types and the potency of inhibition was dependent of the accessibility of the specific LXA(4) receptor, formyl-peptide receptor like-1 (FPRL-1) expressed by all these cells. Immunohistochemistry analysis on human bronchial biopsies showed a high expression of FPRL-1 in the epithelium. Finally, an FPRL-1 receptor antagonist, boc-2 peptide reversed LXA(4) effect on IL-8 generation. Together, these findings indicate that differentiated human bronchial epithelium synthesizes LX in vivo which could have autocrine actions through its specific receptor FPRL-1 to promote resolution of airway inflammation. PMID- 17419000 TI - Mechanism of tert-butylhydroperoxide induced cytotoxicity in U-937 macrophages by alteration of mitochondrial function and generation of ROS. AB - tert-Butylhydroperoxide has been reported to inhibit growth and induce apoptosis in number of cell types, but little is known about the molecular mechanism mediating these effects. In the present study, we determined the molecular pathways that lead to apoptosis after treatment of cells with t-BOOH. The cells were exposed to different concentrations of t-BOOH (100-750 microM) for 1-4 h and various parameters such as cytotoxicity, ROS (reactive oxygen species) generation, MMP (mitochondrial membrane potential), intracellular Ca++ levels and expression of various proteins involved in apoptosis were determined. Exposure of U-937 cells to t-BOOH induced cytotoxicity in a time dependent manner with about 50% toxicity at 400 microM t-BOOH in 4h. t-BOOH treatment resulted in a time dependent increase in reactive oxygen species levels, Ca++ influx and annexin V positive cells. There was a significant fall in MMP following exposure to t-BOOH with time. t-BOOH treatment of U-937 cells leads to apoptosis, which is accompanied by activation of caspase-3. The caspase-3 inhibitor (Ac-DEVD-CHO) inhibits the cytotoxicity induced by t-BOOH, indicating a direct link between caspase-3 activation and cell death. This activation of apoptosis is accompanied by release of cytochrome c, down regulation of anti-apoptotic protein Bcl-2 levels with concurrent increase in pro-apoptotic proteins Bax and Bad levels. These observations indicate that t-BOOH induces cell death in U-937 macrophages by apoptosis, which is mediated through mitochondrial pathway. PMID- 17419001 TI - Prospective open-label pilot trial of mirtazapine in children and adolescents with social phobia. AB - Mirtazapine is indicated for major depression and used for anxiety in adults; however, little is known about its application in pediatric populations. This is an 8-week open-label pilot study of mirtazapine in children with social phobia age 8-17 years. Primary outcomes were symptom improvement based on clinician rating and self-report, as well as tolerability based on rates of discontinuation due to adverse effects. Fifty-six percent (10/18) responded to treatment, 17% (3/18) achieved full remission. Social phobia symptoms improved significantly during the first 2 weeks of treatment, as did comorbid symptoms of depression and anxiety. Eleven patients (61%) did not complete all 8 weeks of treatment; four patients (22%) discontinued due to adverse effects including fatigue and irritability. The others discontinued due to study burden (28%), insufficient response (6%), or to pursue herbal treatment (6%). Significant weight gain was observed. Larger controlled trials are needed to further evaluate efficacy and safety. PMID- 17419002 TI - Further examination of the relationship between anxiety sensitivity and depression: the mediating role of experiential avoidance and difficulties engaging in goal-directed behavior when distressed. AB - This study examined the role of experiential avoidance and difficulties engaging in goal-directed behavior when distressed in the relationship between anxiety sensitivity (AS) and depressive symptom severity. A sample of 391 undergraduate students completed a series of questionnaires assessing the constructs of interest. Results provided support for a model where experiential avoidance and difficulties engaging in goal-directed behavior mediate the relationship between the AS dimensions of fear of cognitive dyscontrol and fear of publicly observable anxiety reactions and depressive symptom severity. The ability of this model to distinguish participants (N=53) reporting clinical levels of depression from those without (N=53) was then examined. The model was found to reliably distinguish between participants with and without clinical levels of depression. However, only experiential avoidance was a significant mediator. Implications for research on the role of AS in depression vulnerability and treatment are discussed. PMID- 17419003 TI - Distal revascularization and interval ligation (DRIL) procedure for the treatment of ischemic steal syndrome after arm arteriovenous fistula. AB - The objective of this study was to investigate the efficacy of the distal revascularization-interval ligation (DRIL) technique in alleviating symptoms of ischemic steal syndrome and in preserving hemodialysis access. A retrospective chart review was conducted of all patients receiving the DRIL procedure in a 3 year period. There were 38 DRIL bypass grafts identified in 35 patients, with 36 DRILs with follow-up adequate for analysis. Comparison of preoperative and postoperative digital pulse volume recording (PVR) data was made using the t test. The majority of patients presented with multiple ischemic symptoms, most commonly coolness, pain, and paresthesias. Six patients presented with frank digital necrosis. The mean interval to DRIL was 4.9 months following fistula construction (range 0.1-24). In 66.7% of patients for whom complete follow-up data were available (24/36), all ischemic symptoms were alleviated by DRIL. Of the remaining 12 patients, 11 experienced partial symptom relief. One patient required digital amputation following DRIL. A comparison of pre- and post-DRIL PVRs illustrated a significant increase in these values following DRIL (P < 0.05). DRIL effectively eliminates ischemic symptoms in the majority of patients and produces a significant increase in flow to the ischemic limb. The data support the usage of DRIL as the procedure of choice in the correction of ischemic steal following arm arteriovenous fistula. PMID- 17419004 TI - Multiple digital gangrene after a radial artery penetrating injury: a case report. AB - We describe the case of a young female patient who sustained a radial artery penetration injury from broken glass after falling and was managed only by external compression without arterial repair. Although the patient did not have any reported predisposing factors and the radial artery was normal as well as patent in the angiographic examination, she developed progressive multiple digital ischemia and required subsequent amputation of the gangrenous fingers. The possible mechanism might be an occlusion of the digital arteries originating from the injured radial artery by dispersion of thrombembolic particles distally during the injury as there was sparing of involvement of the ring finger, which wore a ring that restricted the thromboembolic particles from passing distally during the accident. This case revealed that particular attention should be paid to those patients with a radial artery penetration injury to facilitate early detection and implementation of therapy for possible finger ischemia and subsequent gangrene. PMID- 17419005 TI - Experimental study of a new vascular anastomotic technique in a swine model: short and mid-term results. AB - This study reports the development of a new open external vascular stent: the lock stent device (LSD). It enables a novel end-to-end sleeve anastomotic technique, named locked anastomosis (LA). The LA technique consists of inserting a graft sleeve within an artery, secured in place by an internal stent and an external LSD. The internal stent, graft, vessel wall, and LSD are fastened together with single sutures. The LSD placement does not require a complete transection of the vessel wall as it can be enlarged and then relaxed to clasp the vessel neck. The tensile strength of the LA technique was accurately measured ex vivo and its stability successfully tested in vivo by acute pressure peak tests and mid-term survival studies. Ex vivo, the ratio between the tensile strength of the LA technique with two, three, and six stitches and that of hand sewn anastomoses was 0.41 +/- 0.02, 0.59 +/- 0.17, and 1.03 +/- 0.04. In vivo, LA anastomoses tolerated marked increases in blood pressure (peak systolic pressure 195-230 mm Hg) for periods of 15-25 min without leakage. Five pigs survived 10 weeks with abdominal aorto-aortic bypass performed according to the LA technique with three stitches. Aortograms showed no narrowing or thromboses, and histological findings confirm uniform flattening of the aortic wall at the anastomosis, with proliferating neointima and uniformly hypotrophic media. Minimal changes were observed in the adventitia. PMID- 17419006 TI - Follow-up and treatment of nutcracker syndrome. PMID- 17419007 TI - Complement activation in anti-phospholipid syndrome: a clue for an inflammatory process? AB - Anti-phospholipid syndrome (APS) is defined by recurrent arterial/venous thrombosis and/or fetal losses in the persistent presence of anti-phospholipid antibodies (aPL). In in vivo experimental models aPL thrombogenic activity is associated with a pro-inflammatory endothelial phenotype (increased adhesion molecule [ADM] expression and leukocyte adhesion) in addition to a pro-coagulant one (tissue factor [TF] expression). This is in line with the in vitro aPL ability to trigger intracellular signalling and to up-regulate ADM, TF and pro inflammatory cytokine/chemokine expression at the mRNA and protein level in endothelial cells. Comparable effects were also reported in monocytes in vitro. In addition, complement activation is required by aPL to display their thrombogenic activity in in vivo models. Interestingly, complement activation blocking as well as Tumor Necrosis Factor alpha neutralization protect animals from aPL-induced fetal losses. Altogether these findings speak in favour for a role of inflammation in APS in spite of the absence of a clear inflammatory signature in the patients. We could not find any complement (C3c and C4d) deposition in the placentas from 2 late abortions (20 weeks of gestation) in APS women. Further studies are necessary to investigate whether complement activation and inflammatory processes found in animal models are taking place in APS patients. PMID- 17419008 TI - Antipsychotic efficacy: relationship to optimal D2-receptor occupancy. AB - Clinically important differences exist between antipsychotic agents and formulations in terms of safety and tolerability. Features of the biochemical interaction between the antipsychotic and the D2-receptor may underlie these differences. This article reviews current information on the relationship between antipsychotic receptor occupancy and clinical response. A literature search was performed using the keywords 'antipsychotic or neuroleptic', 'receptor' and 'occupancy' and 'dopamine' and 'D2' supplemented by the authors' knowledge of the literature. Imaging and clinical data have generally supported the hypotheses that optimal D2-receptor occupancy in the striatum lies in a 'therapeutic window' between approximately 65 and approximately 80%, however, pharmacokinetic and pharmacodynamic properties of a drug should also be taken into account to fully evaluate its therapeutic effects. Additional research, perhaps in preclinical models, is needed to establish D2-receptor occupancy in various regions of the brain and the optimal duration of D2-receptor blockade in order to maximise efficacy and tolerability profiles of atypical antipsychotics and thereby improve treatment outcomes for patients with schizophrenia. PMID- 17419009 TI - Role of the COMT gene Val158Met polymorphism in mental disorders: a review. AB - The Val158Met polymorphism of the COMT gene is functional, easily detectable, and significantly related to metabolism of catecholamines, which underlie pathogenesis of a significant number of mental disorders. Evidence for the role of this polymorphism in schizophrenia, substance dependence, bipolar disorder, obsessive-compulsive disorder, anorexia nervosa and attention deficit hyperactivity disorder is summed up in this review article. The results make it unlikely that the COMT gene plays an important role in these mental disorders, although a minor effect can not be excluded. Future studies on the COMT gene in mentally ill subjects should be stratified by clinical subtypes of the disorder, gender and ethnicity. Studies of endophenotypes instead of the complex disorder seem to be another promising research strategy. Gene-gene and gene-environment interactions should also be considered. The COMT gene is probably not "a gene for" any mental disorder, but the Val158Met polymorphism appears to have pleiotropic effects on human behavior. PMID- 17419010 TI - Serotonergic response to stress: a protective factor against abnormal dopaminergic reactivity in schizophrenia? AB - In a double-blind-crossover-study, 10 patients with schizophrenia, 10 of their unaffected siblings and 9 healthy controls randomly received metabolic stressor and placebo. A significant HVA plasma elevation in response to stress was found in siblings whose response was intermediate to that of patients and controls. Only siblings additionally displayed an exaggerated 5HIAA response. PMID- 17419011 TI - Role of surfactants in clouding phenomenon of imipramine hydrochloride. AB - The clouding behavior of tricyclic antidepressant drug imipramine hydrochloride (IMP) in aqueous solution has been studied in presence of surfactants. A pH increase in the presence as well as in the absence of surfactants decreased the CP. Drug molecules become neutral at high pH and therefore head-head repulsion decreases which lead to CP decrease. Addition of non-ionic and cationic surfactants increased the CP whereas anionic surfactants showed a peaked profile. Effect of CTAB/TX-100 at different fixed drug concentrations showed that at all surfactant concentrations the CP value was higher for higher drug concentrations. However, variation of pH produced opposite effect: CP at all CTAB/TX-100 concentrations decreased with increasing pH. All results are interpreted in terms of increase in hydrophobicity or hydrophilicity of micelles on addition of surfactants. PMID- 17419012 TI - Preparation and characterization of temperature and pH-sensitive chitosan material and its controlled release on coenzyme A. AB - A novel copolymer P(CS-Ma-DMAEMA) was synthesized with chitosan (CS), maleic anhydride (Ma) and 2-(dimethylamino)ethyl methacrylate (DMAEMA) by grafting and copolymerization. The copolymer obtained was analyzed by FT-IR, (1)H NMR and UV, and the molecular weight and polydispersity were determined by gel permeation chromatography (GPC). The average size and distribution of copolymer micelles were determined by dynamic light scattering (DLS). Their aqueous solution properties and controlled coenzyme A delivery were also studied. It was found that the copolymer had temperature sensitivity and pH sensitivity. The factors affecting release behavior, such as concentration, pH and temperature were discussed in this paper. The higher concentration of the copolymer aqueous solution absorbed more coenzyme A than the lower one. The increasing temperature accelerated the drug release from the copolymer. The pH of the copolymer solution had significant impact on the release of coenzyme A. The results suggested that the novel copolymer could be used as drug delivery carrier. PMID- 17419013 TI - [A diagnostic approach for inherited thrombocytopenia]. PMID- 17419014 TI - [Reference center for platelet diseases]. PMID- 17419015 TI - [Hypnosis at the paediatric emergency ward]. PMID- 17419016 TI - [Sedation in the emergency department: how far can and should we go?]. PMID- 17419017 TI - [Eating disorders and later obesity risk]. PMID- 17419018 TI - [Late effects of cancer chemotherapy in children]. PMID- 17419019 TI - [Focus on difficulties encountered by gifted children]. PMID- 17419020 TI - [School-based nutrition programs and behaviour: what should we expect?]. PMID- 17419021 TI - [Oxytocin and the central nervous system]. PMID- 17419022 TI - [The transition in cystic fibrosis care: an adult physician's point of view]. PMID- 17419023 TI - [Clinical research in pediatric oncology department: children and parents opinion]. PMID- 17419024 TI - [Possible long-term effect of in utero antiretroviral exposure?]. PMID- 17419025 TI - [Palliative care in pediatrics: myth of fact?]. PMID- 17419026 TI - [Emerging viral diseases in pulmonary medicine]. PMID- 17419027 TI - [Consequences in adulthood of childhood obesity]. PMID- 17419028 TI - [The truth disclosure in paediatric oncology]. PMID- 17419029 TI - [Transition from paediatric to adult care for diabetic adolescents: to be or not to be lost in translation?]. PMID- 17419030 TI - [The adoption in a generous society and a kind family]. PMID- 17419031 TI - [Perinatal consequences of substance abuse during pregnancy]. PMID- 17419032 TI - [Renal hypodysplasia]. PMID- 17419033 TI - [The course, steps of the parents for adopting a child]. PMID- 17419034 TI - [Transition care in pediatric rheumatology]. PMID- 17419035 TI - [Rapid test performance characteristics evaluation]. PMID- 17419036 TI - [The disorders of attachment in adoption]. PMID- 17419037 TI - [Classification of angiomas]. PMID- 17419038 TI - A case of hypersensitivity vasculitis after vinorelbine injection in a patient with metastatic breast cancer. PMID- 17419039 TI - Fibromatosis: benign by name but not necessarily by nature. AB - Aggressive fibromatoses, also known as desmoid tumours, are rare fibrous tissue proliferations with a tendency for slow, local infiltrative growth. There is an association with Gardner's syndrome and familial adenomatous polyposis. Histologically they are fairly bland with no abnormal mitoses or necrosis. They do not metastasize, but can cause significant morbidity through their locally destructive effects. Magnetic resonance imaging is the method of choice for diagnosis, pre-treatment planning and post-treatment follow-up. Surgical excision with a wide margin is the treatment of choice. However, there is a tendency for local recurrence and repeated excision may result in a poor functional or cosmetic outcome. Radiotherapy is used to reduce local recurrence rates after excision and is also used to treat inoperable tumours. Long-lasting remissions can be obtained. Treatment is now planned using modern three-dimensional conformal techniques, similar to those used in soft tissue sarcoma management. There is no definite dose-response relationship, but doses of 50-60 Gy in 1.8-2 Gy fractions are recommended. Systemic therapy has been used for lesions not controlled by surgery or radiotherapy, or less commonly, as a primary treatment. Tamoxifen and non-steroidal anti-inflammatory agents are used most often as they are relatively non-toxic, but there is limited experience with cytotoxic chemotherapy and biological agents. There are no randomised trials to help guide the management of this locally aggressive 'benign' tumour and treatment decisions are best made by the local soft tissue sarcoma multidisciplinary team. PMID- 17419040 TI - The genomics revolution and radiotherapy. AB - The expansion of our knowledge through the Human Genome Project has been accompanied by the development of new high-throughput techniques, which provide extensive capabilities for the analysis of a large number of genes or the whole genome. These assays can be carried out in various clinical samples at the DNA (genome), RNA (transcriptome) or protein (proteome) level. There is a belief that this genomic revolution, i.e. sequencing of the human genome and developments in high-throughput technology, heralds a future of personalised medicine. For clinical oncology, this progress should increase the possibility of predicting individual patient responses to radiotherapy. This review highlights some of the work involving sparsely ionising radiation and the new technologies. PMID- 17419041 TI - The use of fluorescein for labeling genomic probes in the checkerboard DNA-DNA hybridization method. AB - Molecular methods that permit the simultaneous detection and quantification of a large number of microbial species are currently employed in the evaluation of complex ecosystems. The checkerboard DNA-DNA hybridization technique enables the simultaneous identification of distinct bacterial species in a large number of dental samples. The original technique employed digoxigenin-labeled whole genomic DNA probes which were detected by chemiluminescence. In this study, we present an alternative protocol for labeling and detecting whole genomic DNA probes in the Checkerboard DNA-DNA hybridization method. Whole genomic DNA was extracted from five bacterial species and labeled with fluorescein. The fluorescein labeled whole genomic DNA probes were hybridized against whole genomic DNA or subgingival plaque samples in a checkerboard hybridization format, followed by chemiluminescent detection. Our results reveal that fluorescein is a viable and adequate alternative labeling reagent to be employed in the checkerboard DNA-DNA hybridization technique. PMID- 17419042 TI - Reference values for dynamic and static pulmonary compliance in men. AB - The aim of the present study was to determine new reference values and predictive variables for dynamic and static pulmonary compliance in men. The investigation was conducted as a prospective study in healthy, non-smoking men with normal pulmonary function parameters including spirometry, bodyplethysmography and CO diffusing capacity. The esophageal pressure method was used to measure dynamic compliance (Cdyn), specific dynamic compliance (Cdyn/ITGV), static compliance (Cstat) and specific static compliance (Cstat/ITGV). Lung recoil pressures were recorded at different levels of total lung capacity (TLC). A total of 208 men aged 20-69 years were included in the study. The mean values for the compliance parameters were: Cdyn: 2.91+/-1.08 L/kPa; Cdyn/ITGV: 0.71 +/- 0.30 kPa (-1); Cstat: 3.34 +/- 1.04 L/kPa; Cstat/ITGV: 0.82 +/- 0.31 kPa (-1). Cdyn, Cdyn/ITGV and Cstat/ITGV were significantly correlated with age and Cstat was related to height, but in multiple regression analyses the predictability for compliance parameters was very low. Lung recoil pressures at all TLC levels significantly decreased with ageing. In conclusion, we demonstrated that the contribution of anthropometric variables to the regression equations of pulmonary compliance was low. With ageing the static pressure-volume curve of the lung shifted to the left without substantial alteration of the slope. PMID- 17419043 TI - Cost analysis of GER-induced asthma: a controlled study vs. atopic asthma of comparable severity. AB - Bronchial asthma is a costly disease: while the role of pharmaceutical strategies was greatly emphasised in order to alleviate its economic burden, the aetiological approach to asthma has received much less attention from this point of view. The impact of gastro-oesophageal reflux (GER)-related asthma was assessed in comparison to atopic asthma in 262 matched patients, and the corresponding direct and indirect annual costs calculated. All subjects were screened by means of a 95-item self-questionnaire. The overall resource utilisation was calculated for the last 12 months. Drug-induced annual costs were euro 290.4 (interquartile range-iqr 32.8) in atopic and euro 438.4 (iqr 27.8) in GER-related asthma (p<0.001); expenditure for medical consultations and diagnostics were euro 166.1 (iqr 14.8) vs. euro 71.6 (iqr 11.0) (p<0.001), and euro 338.4 (20.0) vs. 186.9 (iqr 26.5) (p<0.001), respectively. Direct costs due to hospital admissions and indirect costs due to absenteeism were also higher in GER-related asthmatics: 2.201.7+/-90.0 vs. euro 567.1+/-11.0 (p<0.001), and euro 748.7+/-94.7 vs. euro 103.6+/-33.9 (p<0.001), respectively. The total annual cost per patient was euro 1246.7 (iqr 1979.6) in atopic and euro 3967.1 (iqr 3751.5) in GER-related asthma, p<0.001. In conclusion, GER-induced asthma has a more relevant economic impact on healthcare resources than atopic asthma. Although further studies are needed, present data tend to demonstrate that when facing difficult asthma (GER-related asthma in this case), the aetiological assessment of the disease plays a critical role in optimising the approach to patients' needs. PMID- 17419044 TI - Salinity, organic content, micronutrients and heavy metals in pig slurries from South-eastern Spain. AB - The increase in commercial pig production is an opportunity to reuse animal manures in arid and semiarid soils as a source of nutrients and organic matter. However, there are components in pig slurry that are potentially dangerous for the environment. In this study, pig slurries of 36 pig farms in South-eastern Spain were evaluated for salt content (electrical conductivity, chloride and sodium), organic load (BOD5 and COD), micronutrients (Fe, Cu, Mn and Zn), and heavy metals (Cd, Co, Cr, Ni and Pb). Except for electrical conductivity, Cu and Zn, components in pig slurries did not vary considerably between animal production stages, indicating similar management of diverse animal types. Assuming an application rate based on the maximum input of nitrogen from animal manure (210 kg total N/ha/yr), the estimates for soil annual load of Cl and Na, 415 kg/ha, could be a significant salinisation risk. Cu and Zn seemed to be the metals that could be accumulated most in soils where application of pig slurries is common (4 and 15 kg/ha/yr, respectively). The estimated heavy metal (Cd, Co, Cr, Ni and Pb) input to soils would be 260 g/ha/yr, with a relative contribution of Cr>Ni>Pb>Co>Cd. PMID- 17419045 TI - Life cycle assessment of a national policy proposal - the case of a Swedish waste incineration tax. AB - At the core of EU and Swedish waste policy is the so-called waste hierarchy, according to which waste should first be prevented, but should otherwise be treated in the following order of prioritisation: reuse, recycling when environmentally motivated, energy recovery, and last landfilling. Some recent policy decisions in Sweden aim to influence waste management in the direction of the waste hierarchy. In 2001 a governmental commission assessed the economic and environmental impacts of introducing a weight-based tax on waste incineration, the purpose of which would be to encourage waste reduction and increase materials recycling and biological treatment. This paper presents the results of a life cycle assessment (LCA) of the waste incineration tax proposal. It was done in the context of a larger research project concerning the development and testing of a framework for Strategic Environmental Assessment (SEA). The aim of this paper is to assess the life cycle environmental impacts of the waste incineration tax proposal, and to investigate whether there are any possibilities of more optimal design of such a tax. The proposed design of the waste incineration tax results in increased recycling, but only in small environmental improvements. A more elaborate tax design is suggested, in which the tax level would partly be related to the fossil carbon content of the waste. PMID- 17419046 TI - Development of fructosyl valine binding polymers by covalent imprinting. AB - Molecularly imprinted polymers (MIPs) against fructosyl valine (Fru-Val), the N terminal constituent of hemoglobin A1c beta-chains, were prepared by cross linking of beta-D-Fru-Val-O-bis(4-vinylphenylboronate) with an excess of ethylene glycol dimethacrylate (EDMA) or trimethylolpropane trimethacrylate (TRIM). Control MIPs were prepared in analogy by cross-linking the corresponding vinylphenylboronate esters of fructose and pinacol. After template extraction batch rebinding studies were performed using different pH values and buffer compositions. The Fru-Val imprinted TRIM cross-linked polymer binds about 1.4 times more Fru-Val than the fructose imprinted polymer and 2.7 times more Fru-Val than pinacol imprinted polymer. The highest imprinting effect was obtained in 100 mM sodium carbonate/10% methanol (pH 11.4). The TRIM cross-linked Fru-Val imprinted polymer showed a better specificity than the EDMA cross-linked polymer. The binding of valine was very low. Thermo gravimetric analysis indicated that the generated Fru-Val imprinted polymer has high thermo stability. No change in binding was observed after incubation of the polymers in buffer at 80 degrees C for 36 h. Since the functional group of the polymers (phenyl boronic acid) targets the sugar part of Fru-Val the imprint technique used should also be applicable for the development of MIPs against other glycated amino acids and peptides. PMID- 17419047 TI - Structural aspects of AMPA receptor activation, desensitization and deactivation. AB - Glutamate mediates most of the excitatory neurotransmission in the mammalian central nervous system by activating ionotropic glutamate receptors. Structural and functional studies of ionotropic glutamate receptors have offered detailed insight into the mechanism by which these integral membrane proteins function. In particular, advances in our understanding of the atomic structure of the agonist binding domain have provided new opportunities to consider the conformational changes that take place in a functioning ligand-gated ion channel. Several recent studies have turned up important new ideas about the structural determinants of channel activation, deactivation and desensitization of AMPA receptors. Working hypotheses derived from this structural insight offer a rare opportunity to enrich and guide functional studies. PMID- 17419048 TI - Ca(2+)-assisted receptor-driven endocannabinoid release: mechanisms that associate presynaptic and postsynaptic activities. AB - Endogenous cannabinoids (endocannabinoids) serve as retrograde messengers at synapses in various regions of the brain. They are released from postsynaptic neurons and cause transient and long-lasting reduction of neurotransmitter release through activation of presynaptic cannabinoid receptors. Endocannabinoid release is induced either by increased postsynaptic Ca(2+) levels or by activation of G(q/11)-coupled receptors. When these two stimuli coincide, endocannabinoid release is markedly enhanced, which is attributed to the Ca(2+) dependency of phospholipase Cbeta (PLCbeta). This Ca(2+)-assisted receptor-driven endocannabinoid release is suggested to participate in various forms of synaptic plasticity, including short-term associative plasticity in the cerebellum and spike-timing-dependent long-term depression in the somatosensory cortex. In these forms of plasticity, PLCbeta seems to function as a coincident detector of presynaptic and postsynaptic activities. PMID- 17419049 TI - Neurotrophins, synaptic plasticity and dementia. AB - The growing realization that neurotrophins, such as brain-derived neurotrophic factor (BDNF), are crucial in modulating synaptic plasticity has broadened the spectrum of their trophic actions. At the same time, it has become clear that Abeta peptides derived from amyloid precursor protein (APP) have dramatic effects on synaptic transmission before the onset of the neurodegenerative disease. Because neurotrophins and Abeta are responsible for affecting both synaptic and cognitive function, it is likely that their mechanisms of action will be related and might even intersect. This review highlights several recent findings that suggest trophic factors and APP use similar pathways to control neuronal activity. PMID- 17419051 TI - Investigating biological systems using first principles Car-Parrinello molecular dynamics simulations. AB - Density functional theory (DFT)-based Car-Parrinello molecular dynamics (CPMD) simulations describe the time evolution of molecular systems without resorting to a predefined potential energy surface. CPMD and hybrid molecular mechanics/CPMD schemes have recently enabled the calculation of redox properties of electron transfer proteins in their complex biological environment. They provided structural and spectroscopic information on novel platinum-based anticancer drugs that target DNA, also setting the basis for the construction of force fields for the metal lesion. Molecular mechanics/CPMD also lead to mechanistic hypotheses for a variety of metalloenzymes. Recent advances that increase the accuracy of DFT and the efficiency of investigating rare events are further expanding the domain of CPMD applications to biomolecules. PMID- 17419052 TI - Probabilities for survival of glassy-winged sharpshooter and olive fruit fly pests in urban yard waste piles. AB - Glassy-winged sharpshooter (Homolodisca coagulate) and olive fruit fly (Bactrocera oleae) were introduced into unturned, chipped yard waste piles to evaluate their survival with time and depth within the piles. In all three trials, no pests lasted more than 14 d, and in no trial did pests survive more than 4d at the 30 and 100 cm depths. No survivors were found after 14 d in any of the treatments at any depth. Neither of the pests survived 100 cm after 2d. A mathematical model for describing pest survival probabilities is described. The model modifies time according to the Arrhenius equation in order to include heat effects on pest survival and can be used to determine exposure times necessary to eliminate these pests with a determined statistical probability. Model projections suggest that for conditions similar to this study, there is 99% confidence that all glassy-winged sharpshooter eggs would be eliminated from 1000 infected leaves in 6.1d at 15 cm depth and in 4.8d at 30 cm or below. Olive fruit fly larvae at these depths would require 4.8 and 4.1d, respectively, for 1000 infected olive fruits. Projected elimination times at the surface were longer, 6.5d for sharpshooter eggs and 14.3d for fruit fly larvae. PMID- 17419053 TI - Novel benzodifuran analogs as potent 5-HT2A receptor agonists with ocular hypotensive activity. AB - A series of 8-substituted benzodifuran analogs was prepared and evaluated for 5 HT(2A) receptor binding and activation. Several compounds containing ether and ester functionality were found to be potent agonists. Topical ocular administration of 5, 18, and 25 effectively reduced intra-ocular pressure in the hypertensive cynomolgus monkey eye in the range of 25-37%. PMID- 17419054 TI - Minimum structure requirement of immunomodulatory glycolipids for predominant Th2 cytokine induction and the discovery of non-linear phytosphingosine analogs. AB - Analogs of immunomodulatory glycolipid OCH (2) were prepared and minimum structure requirement to exhibit equivalent profiles was disclosed. Analogs bearing non-linear hydrocarbon chain in the phytosphingosine moiety (18, 19) were shown for the first time to possess comparable cytokine inducing profile to 2. Molecular modeling of 2/hCD1d complex based on the crystal structure of alpha GalCer (1)/hCD1d complex is also described. PMID- 17419055 TI - Identification of 6-substituted 4-arylsulfonyl-1,4-diazepane-2,5-diones as a novel scaffold for human chymase inhibitors. AB - A novel series of 6-substituted 4-sulfonyl-1,4-diazepane-2,5-diones were designed, synthesized and evaluated as human chymase inhibitors. Structure activity relationship studies led to the identification of a potent inhibitor, (6S)-6-(5-chloro-2-methoxybenzyl)-4-[(4-chlorophenyl)sulfonyl]-1,4-diazepane-2,5 dione, with an IC(50) of 0.027 microM. PMID- 17419056 TI - Novel aminoethylbiphenyls as 5-HT7 receptor ligands. AB - The synthesis of a series of aminoethylbiphenyls as novel 5-HT(7) receptor ligands is described. The novel derivatives exhibit high affinity for the 5-HT(7) receptor with selectivity toward 5-HT(1A) receptor. PMID- 17419057 TI - The level of knowledge of respiratory physiology articulated by intensive care nurses to provide rationale for their clinical decision-making. AB - The combination of intensive care knowledge, and the ability to articulate analytical decision-making processes to the interdisciplinary team, enhances the clinical credibility of the intensive care unit (ICU) nurse. The objective of this paper is to outline a study firstly, assessing ICU nurses' ability in articulating respiratory physiology to provide rationale for their clinical decision-making and secondly, the barriers that limit the articulation of this knowledge. Using an evaluation methodology, multiple methods were employed to collect data from 27 ICU nurses who had completed an ICU education programme and were working in one of two tertiary ICUs in New Zealand. Quantitative analysis showed that nurses articulated a low to medium level of knowledge of respiratory physiology. Thematic analysis identified the barriers limiting this use of respiratory physiology as being inadequate coverage of concepts in some ICU programmes; limited discussion of concepts in clinical practice; lack of clinical support; lack of individual professional responsibility; nurses' high reliance on intuitive knowledge; lack of collaborative practice; availability of medical expertise; and the limitations of clinical guidelines and protocols. These issues need to be addressed if nurses' articulation of respiratory physiology to provide rationale for their clinical decision-making is to be improved. PMID- 17419058 TI - Observations of an American nurse. PMID- 17419059 TI - The effect of total knee replacement surgery on gait stability. AB - The purpose of this study was to investigate the effect of knee pain and total knee replacement (TKR) surgery on the gait stability of knee osteoarthritis (OA) patients compared to controls. Gait spatio-temporal measures, center of mass kinematics and pain levels of 21 TKR subjects and 21 controls (CON) were assessed during level walking and obstacle crossing at two testing periods, pre-surgery (P1) and 6 months post-surgery (P2). The TKR patients reported greater pain and disability than CONs at P1 and P2, walked and negotiated the obstacle more slowly and had a shorter stride length than CONs. After surgery, the TKR center of mass center of pressure (COM-COP) separation distance and the peak anterior inclination angle were significantly smaller than CONs. Pain was found to be significantly related to sagittal plane measures, but not to similar measures in the frontal plane. The data suggest that total knee replacement surgery and pain affect gait stability predominantly in sagittal plane variables. The TKR subjects used a conservative strategy to manage the COM and COP in the sagittal plane, possibly to reduce the kinetic demands on the involved limb. PMID- 17419060 TI - Detecting spinal posture change in sitting positions with tri-axial accelerometers. AB - A method of using three tri-axial accelerometers for monitoring postural changes in sitting was introduced. Subjects were requested to perform forward trunk flexion and lateral bending. A motion analysis system and a three-dimensional rotation alignment device were used to verify the measurements of the accelerometers. The precision and reliability of the accelerometers for measuring static calibration was demonstrated as < or =1 degrees for the RMS error and 1.000 for ICC(3,1). For measurements of sitting posture, the averaged RMS difference between the measurements of the accelerometers and the motion analysis system were < or =5 degrees for all sitting postures on the coronal and sagittal planes. The results of this study demonstrated that tri-axial accelerometers could be used for tracking spinal postural changes on the sagittal and coronal planes in the sitting positions. PMID- 17419061 TI - Assessment of foot impairment in rheumatoid arthritis patients by dynamic pedobarography. AB - OBJECTIVES: Impaired foot function may jeopardize the mobility of patients with rheumatoid arthritis (RA). However, there are still no guidelines concerning the adequate early treatment of painful rheumatoid feet which do not yet require surgery. An assessment method for RA feet appears necessary in order to detect foot problems before functional limitations develop. Therefore, the aim of the present study was to evaluate the use of pedobarographic measurements for detecting changes in plantar loading characteristics and their relationship to foot pain in patients with RA. METHODS: One hundred and twelve patients with RA (55.0+/-11.0 years of age) were divided into three groups according to their Health Assessment Questionnaire (HAQ) Score and compared to a control group of 20 healthy adults (CG). Thirty-six patients with good physical capacity belonged to group 1 (RA1; HAQ-score: 0-1.0), 38 patients with moderate capacity to group 2 (RA2; score: 1.1-2.0) and 38 patients with low capacity to group 3 (RA3; score: 2.1-3.0). Each patient's foot pain was clinically assessed. Pedobarography was used to analyze foot loading parameters while walking barefoot. RESULTS: In the forefoot, average pressures under the lateral forefoot were higher in RA1 patients than in RA2 patients and controls (p<0.05) despite an inconspicuous clinical examination of the foot in RA1 patients. RA1 patients also demonstrated higher plantar pressures than RA2 under the second metatarsal head (p<0.05). In contrast, no significant differences in maximum force could be demonstrated between patient groups. Furthermore, in RA3 patients with lower physical capacity, foot pain was increased as compared to RA1 and RA2 patients. CONCLUSION: In RA patients, pedobarographic patterns show specific changes which characterize the level of functional capacity. In patients with foot involvement, pedobarographic measurements can be useful during the earlier stages of the disease, when clinical examination does not yet indicate the need for more aggressive treatment or orthopedic interventions. PMID- 17419062 TI - Life on the edge: a link between gene expression levels and aggregation rates of human proteins. PMID- 17419063 TI - Why are prion diseases precluded by non-mammals? PMID- 17419064 TI - A new class of nifuroxazide analogues: synthesis of 5-nitrothiophene derivatives with antimicrobial activity against multidrug-resistant Staphylococcus aureus. AB - Hospital-acquired methicillin-resistant Staphylococcus aureus (MRSA) has been an increasing problem worldwide since the initial reports over 40 years ago. To examine new drug leads with potential antibacterial activities, 14 p-substituted benzoic acid [(5-nitro-thiophen-2-yl)-methylene]-hydrazides were designed, synthesized, and tested against standard and multidrug-resistant S. aureus strains by serial dilution tests. All compounds exhibited significant bacteriostatic activity and some of them also showed bactericidal activity. The results confirmed the potential of this class of compounds as an alternative for the development of selective antimicrobial agents. PMID- 17419065 TI - Improved biochemical strategies for targeted delivery of taxoids. AB - Paclitaxel (Taxol) and docetaxel (Taxotere) are very important anti-tumor drugs in clinical use for cancer. However, their clinical utility is limited due to systemic toxicity, low solubility and inactivity against drug resistant tumors. To improve chemotherapeutic levels of these drugs, it would be highly desirable to design strategies which bypass the above limitations. In this respect various prodrug and drug targeting strategies have been envisioned either to improve oral bioavailability or tumor specific delivery of taxoids. Abnormal properties of cancer cells with respect to normal cells have guided in designing of these protocols. This review article records the designed biochemical strategies and their biological efficacies as potential taxoid chemotherapeutics. PMID- 17419066 TI - Eyes of male and female Orgyia antiqua (Lepidoptera; Lymantriidae) react differently to an exposure with UV-A. AB - The structural organization of the eyes belonging to 12 winged male and 12 wingless female Orgyia antiqua moths, exposed for 1 h to UV-radiation (lambda(max)=351 nm) of 1.4 kW/m2, was compared with that of 12 male and 12 female non-irradiated control specimens. Following the UV-exposure, the screening pigments were found in a position indicative of extreme light-adaptation. Extensive formations of vesicles along the perimeter of the cones as well as disintegrating ER in the cone cytoplasm were noticeable, especially in the eye of the female. On the retinal side of the clearzone, the microvilli of the rhabdoms had become affected by the UV in characteristic ways: in the male eye, retinal cell damage in the form of microvillar swellings and disintegrations were largely confined to just two cells per ommatidium, placed opposite to each other. The female eye, once again, exhibited greater vulnerability and more widespread microvillar disruptions that affected all of the ommatidial retinula cells. The greater resistance of the eye of the male to an exposure with UV makes sense, if we consider the consequences of the retinal damage, which would clearly be a more severe handicap for an actively flying individual than for an almost sedentary one like the wingless female. PMID- 17419067 TI - gamma-ray spectrometry of soil samples from the Provincia dell'Aquila (Central Italy). AB - In the Abruzzo Region (Central Italy) there is a lack of measurements of gamma ray activity in soils and waters. For this reason, we have planned to carry out a systematic measurement of soils in the area of the Provincia dell'Aquila, which covers about one-half of the entire region. In this paper we report the results obtained from 56 soil samples, collected in the northern part of the area of interest (about one-fourth of the total area under study). The results, in terms of content of uranium, thorium and potassium and the activity of caesium are reported, as well as the details on the experimental procedure. The results show a limited content of K and U, with no large variations from site to site, in agreement with the expectations based on the knowledge of the geo-lithological nature of the soil. The amount of Th is also quite limited, with a few exceptions where the Th content is up to five times the average value. Caesium, originated from the fall-out following the Chernobyl accident, is very irregularly distributed owing to the complicated orography of the land. Future plans are also shortly discussed. PMID- 17419068 TI - Influence of a tilting prosthetic mitral valve orientation on the left ventricular flow - an experimental in vivo magnetic resonance imaging study. AB - OBJECTIVE: Orientation-related monoleaflet mechanical valve flow and velocity studies in the downstream are limited in mitral valve replacement studies. METHODS: In five sheep, ventricular blood flow was visualized prior to the implantation of a Medtronic Hall tilting valve model. In six sheep, the implant orientation was either anatomical (disc aligned with the anterior leaflet) or anti-anatomical. The mitral subvalvular apparatus was preserved. Sheep were positioned within an 1.5 T field strength MR scanner (Magnetom Sonata; Siemens) to assess time-dependent three dimensional blood flow. RESULTS: The preoperative ventricular velocity profiles presented negligible individual variances. Streamlines passed homogeneously without any spatial differences in flow velocities into the left ventricle. Starting from the anatomical position, blood entered mainly through the major orifice of the mechanical valve. The single artificial leaflet mimicked the rudder effect of the natural anterior mitral leaflet, preventing blood streaming directly towards the septum. The area with inhomogeneous blood velocities in the ventricle increased but not significantly from the preoperative status. The non-axial inflow not directed directly to the apex converted to a similar helix as observed in the preoperative cases. Anti anatomical orientation of the prosthesis caused a significant increase in turbulence immediately after passing the mitral prosthesis. The main stream was changed so significantly that the blood flow shifted towards the septum and caused higher velocities of the stream profiles and turbulence apically. CONCLUSIONS: To achieve optimal hemodynamics, orientation of the mitral tilting valve has to be considered carefully, as has been long known from aortic valve replacement studies. PMID- 17419069 TI - Midterm follow-up of the status of Gore-Tex graft after extracardiac conduit Fontan procedure. AB - OBJECTIVE: Extracardiac conduit Fontan procedure has some theoretical advantages over other types of Fontan procedures, such as optimized flow dynamics, a lower frequency of arrhythmias, and technical ease of procedure. However, lack of growth potential and thrombogenicity of the artificial conduit is the main concern and can possibly lead to reoperation for the conduit stenosis. In this study, we investigated the change and the status of the Gore-Tex graft used in extracardiac conduit Fontan procedure. METHODS: Between 1996 and 2005, 154 patients underwent extracardiac conduit Fontan procedure using Gore-Tex graft. Among these, 46 patients underwent cardiac catheterization during follow-up period. We measured the internal diameter of the conduit and inferior vena cava angiographically. RESULTS: Mean follow-up duration was 36.1+/-19.7 months. The conduit diameter used was 16 mm in 10 patients, 18 mm in 16, 20 mm in 14, 22 mm in 4, and 24 mm in 2 patients. The mean conduit-to-inferior vena cava cross sectional area ratio was 1.25+/-0.33. According to the conduit size used, this ratio was 1.03+/-0.17 for 16 mm conduits, 1.33+/-0.37 for 18 mm, 1.33+/-0.36 for 20 mm, 1.28+/-0.26 for 22 mm, and 1.05+/-0.06 for 24 mm conduits (p<0.05, 16 mm vs 18 mm and 20 mm). The mean percent decrease of the conduit cross-sectional area was 14.3+/-8.5%, and this did not differ significantly according to the conduit size (p=0.82). Follow-up duration and the percent decrease of the conduit cross-sectional area did not show significant correlation (r=0.22, p=0.14). There was no reoperation due to conduit stenosis. CONCLUSIONS: During midterm follow-up of about 3 years, the conduit cross-sectional area decreased by 14%, and this did not differ according to the conduit size used. The extent of decrease of the conduit cross-sectional area remained stable irrespective of the follow-up duration. Sixteen millimeters conduit showed no evidence of clinically significant stenosis, but careful follow-up is warranted because of the possible conduit stenosis relative to the patients' somatic growth. PMID- 17419071 TI - Expression, purification, and characterization of recombinant Metarhizium anisopliae acid trehalase in Pichia pastoris. AB - The mature peptide of Metarhizium anisopliae acid trehalase (ATM1) (EC3.2.1.28) was successfully expressed in Pichia pastoris at high levels under the control of AOX1 promoter. The recombinant ATM1 (reATM1) was secreted into culture medium. After 48-h 0.5% methanol induction, the activity of reATM1 in the culture supernatant reached the peak, 5.35 U/mg. Enzyme with a histidine sequence appended to the C terminus was still active and was purified using metal-chelate affinity chromatography. The yield of purified reATM1 was 2.5 mg from 1L supernatant. The purified reATM1 exhibited a molecular mass of approximately 170 kDa on SDS-PAGE. The optimum temperature and pH of reATM1 were 30 degrees C and 6.0, respectively, and the K(m) and V(max) values for reATM1 were 2.6 mM and 0.305 mmol/min/mg, respectively. Studies showed that the enzymatic properties of reATM1 were similar to those of the native ATM1. PMID- 17419072 TI - The active ring-like structure of SecA revealed by electron crystallography: conformational change upon interaction with SecB. AB - SecA is a multifunctional protein involved in protein translocation in bacteria. The structure of SecA on membrane is dramatically altered compared with that in solution, accompanying with functional changes. We previously reported the formation of a novel ring-like structure of SecA on lipid layers, which may constitute part of the preprotein translocation channel. In the present work, two dimensional crystallization of Escherichia coli SecA on lipid monolayers was performed to reveal the structural details of SecA on lipid layers and to investigate its function. The 2D crystals composed of ring-like structures were obtained by specific interaction between SecA and negatively charged lipid. The 2D projection map and 3D reconstruction from negative stained 2D crystals exhibited a distinct open channel-like structure of SecA, with an outer diameter of 7 nm and an inner diameter of 2 nm, providing the structural evidence for SecA importance in forming the part of the translocation channel. This pore structure is altered after transferring crystals to the SecB solution, indicating that the lipid-specific SecA structure has the SecB binding activity. The strategy developed here provides a promising technique for studying structure of SecA complex with its ligand on membrane. PMID- 17419073 TI - Variability of successive contractions subtracted from unfused tetanus of fast and slow motor units. AB - Stimulation of motor units (MUs) with repeated pulses evokes tetanic contractions, which consist of overlapping mechanical responses. The summation of these responses into tetanus is a nonlinear process due to the dynamic changes in the amplitudes and time parameters of the successive components. In order to study these changes, two MUs (one fast and one slow) of rat medial gastrocnemius muscle were stimulated with a progressively increasing number of pulses, from one (i=1) to sixteen (i=16) at a frequency of 15 Hz for the slow MU and 60 Hz for the fast MU. The individual responses were calculated by subtracting the (i)th from the (i+1)th tetanus recording. The contractions obtained following subtraction were modeled using a novel 6-parameter analytical function. The main conclusions of this study are (1) the newly presented analytical function is able to precisely describe the variable shape of all subtracted experimental contractions; (2) the shapes of successive contractions are variable and the subtracted contractions differ from the individual twitches; (3) as the pulse number increases, the parameters of the subtracted contractions change in a different manner for the slow and fast MUs: for the slow MU, the maximal forces and the time parameters increase considerably up to the 4th response, after which they remain nearly constant or show only a slight increase; for the fast MU, the maximal forces and durations also increase, whereas the remaining time parameters initially increase and then maintain a constant level or decrease, which explains the sag phenomenon visible in the unfused tetanus of fast MUs. PMID- 17419074 TI - Phylogeny of "core Gruiformes" (Aves: Grues) and resolution of the Limpkin Sungrebe problem. AB - Opinions on the systematic relationships of birds in the avian order Gruiformes have been as diverse as the families included within it. Despite ongoing debate over monophyly of the order and relationships among its various members, recent opinion has converged on the monophyly of a "core" group of five families classified as the suborder Grues: the rails (Rallidae), the cranes (Gruidae), the Limpkin (Aramidae), the trumpeters (Psophiidae), and the finfoots (Heliornithidae). We present DNA sequence data from four mitochondrial (cytochrome b, 12S rRNA, Valine tRNA, and 16S rRNA) and three nuclear loci (intron 7 of beta-fibrinogen, intron 5 of alcohol dehydrogenase-I, and introns 3 through 5 of glyceraldehyde-3-phosphate dehydrogenase) to test previous hypotheses of interfamilial relationships within Grues, with particular attention to the enigmatic family Heliornithidae. Separate and combined analyses of these gene sequences confirm the monophyly of Grues as a whole, and of the five families individually, including all three species of Heliornithidae. The preferred topology unambiguously supports relationships among four of the five families, with only the position of Psophiidae remaining equivocal. Bayesian "relaxed-clock" dating methods suggest that the divergences of the three heliornithid species occurred in the mid-Tertiary, suggesting that their present disjunct pantropical distribution is a result of early- to mid-Tertiary dispersal. PMID- 17419075 TI - Estrogen receptor alpha genotype is associated with a reduced prevalence of radiographic hip osteoarthritis in elderly Caucasian women. AB - PURPOSE: This study evaluated the association between polymorphisms in the estrogen receptor (ER) alpha gene (ESR1) and prevalent and incident radiographic hip osteoarthritis (RHOA) in a large, well-defined prospective cohort of elderly Caucasian women. METHODS: Prevalent and incident RHOA was evaluated from all available pelvis X-rays obtained from the Study of Osteoporotic Fractures at baseline and after a mean of 8.3 years. Evaluable DNA samples were available from 4746 of these subjects. RHOA cases were defined by published methods. The ESR1 polymorphisms at intron I (Pvu II for a T/C substitution and Xba I for an A/G substitution) were genotyped in the context of a multiplex polymerase chain reaction (PCR) amplification followed by allele-specific single nucleotide polymorphism (SNP) detection with immobilized oligonucleotide probes in linear arrays. Multiple logistic regression was performed to estimate odds ratios (ORs) and 95% confidence intervals (95% CI) associated with the T/C and A/G polymorphisms. RESULTS: RHOA was present in 12.1% of subjects, of whom 325 had joint space narrowing (JSN) score > or =3 and 130 had an osteophyte score > or =2 and JSN score > or =2. There was a significant reduction in the odds of prevalent RHOA for individuals with the C/C compared to T/T genotype at the Pvu II site with an OR of 0.71 (95% CI: 0.55-0.92) (P=0.01). Adjustments for age, weight, height, hip Bone mineral density (BMD) and estrogen use did not alter the relationship between the C/C genotype and reduced risk of RHOA, with an OR of 0.71 (95% CI: 0.54-0.94) (P=0.01). The risk of incident RHOA was reduced for the Pvu II C/C compared to the T/T genotype (P=0.11). Also, the reduced risk of incident RHOA in C/C subjects varied by estrogen use. There was no association between the Xba I G/G or G/A genotypes and RHOA with OR of 0.82 (95% CI: 0.61 1.10) (P=0.19) compared to women with A/A genotype. CONCLUSIONS: We conclude that the C/C genotype of the ER alpha Pvu II polymorphism was associated with a modestly reduced risk of prevalent and incident RHOA in elderly Caucasian women. Additional work is required to understand how the intron I ESR1 polymorphism may alter joint degeneration. PMID- 17419076 TI - Endothelial cell and macrophage regulation of vascular smooth muscle cell calcification modulated by cholestane-3beta, 5alpha, 6beta-triol. AB - The cellular and molecular mechanisms that mediate vascular calcification remain poorly understood. In our previous study, oxysterol cholestane-3beta, 5alpha, 6beta-triol (Triol) was shown to promote vascular smooth muscle cells (VSMCs) calcification. In this study, by using direct coculture, non-contact transwell coculture, and culture with conditioned media, we investigated the roles of endothelial cells (ECs) and macrophages in the regulation of VSMCs calcification in the absence or presence of Triol. In vitro calcification was induced by incubation of VSMCs with beta-glycerophosphate. The results showed that ECs inhibited VSMCs calcification, as manifested by the reduction of calcium deposition in extracellular matrix. This effect of ECs on calcification was via the secreted soluble factors. Furthermore, the stimulation of ECs by Triol had no influence on ECs inhibition of calcification. On the other hand, macrophages promoted VSMCs calcification via the secreted soluble factors such as reactive oxygen species, which was further enhanced by Triol. Our results supported the roles for ECs and macrophages in vascular calcification, modulated by oxysterols in atherosclerotic plaque. PMID- 17419077 TI - Experimental study of a novel suture-less aortic anastomotic device. AB - OBJECTIVES: To assess the suitability and safety of a novel suture-less hermetic docking head (HDH) device for aortic anastomosis, and to test it in vitro, on cadavers, and in animals. DESIGN: Uncontrolled prospective studies. MATERIALS: HDH device with commercial vascular grafts. METHODS: Two experimental series were conducted in vitro to test the hermetic seal of the anastomosis and the fixation of the device. Another series was performed in 6 cadavers with atherosclerotic aortas. Two in vivo trials tested the HDH device when implanted in ten and five pigs, respectively, with follow-up of 22 weeks. Pathological, histological and radiographic studies of the aorta were performed. RESULTS: The in vitro and cadaveric studies demonstrated the resilience of the device under physiological and extreme conditions (up to 1200mmHg pulsatile flow on the bench and up to 270mmHg in cadaveric studies). These studies also established the safety and ease of application of the device in both normal and atherosclerotic aortas. Insertion was easy and rapid. There was no indication of leakage, narrowing or stenosis at the anastomotic sites. The implants' position was maintained without distal or proximal migration. Reliable fixation was observed despite significant increase (up to 112%) in the weight of the animals. Histologically, normal tissue reaction of the lamina was observed. CONCLUSIONS: The suture-less HDH device's behavior in vitro and in vivo suggests that this implant is useful for aortic anastomosis, and its use is faster and simpler than common suturing techniques. PMID- 17419078 TI - A one-year case series of arthrocentesis with hyaluronic acid injections for temporomandibular joint osteoarthritis. AB - OBJECTIVE: The present study presents a case series on the efficacy of arthrocentesis with hyaluronic acid injections for the treatment of temporomandibular joint osteoarthritis by providing patient evaluations at a one year follow-up. STUDY DESIGN: Twenty-five patients with a diagnosis of osteoarthritis according to the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD axis I group IIIb) underwent a cycle of 5 arthrocenteses with injections (1 per week) of 1 mL hyaluronic acid. A number of clinical parameters (pain at rest and mastication, masticatory efficiency, maximum nonassisted and assisted mouth openings, functional limitation, subjective efficacy, and tolerability of the treatment) were assessed by the same blinded operator at the time of the diagnosis (baseline), at each appointment during the treatment, and at 1-week, 1-month, 3-month, 6-month, and 1-year follow up appointments. RESULTS: Descriptive analysis showed improvements which were maintained over time for all the study parameters. Permutation tests evidenced the significance of changes which occurred in many clinical parameters within the first 2 injections. Differences with baseline levels remained significant at the end of the follow-up period, particularly for the masticatory efficiency and pain at mastication (minimum and maximum) parameters. CONCLUSIONS: Data from the present investigation support findings from studies on other joints, which show the efficacy of serial injections of hyaluronic acid after arthrocentesis to reduce symptoms of osteoarthritis and to maintain improvements over time. PMID- 17419079 TI - RNA interference for subacute sclerosing panencephalitis. PMID- 17419081 TI - Strabismus might be a risk factor for amblyopia recurrence. AB - BACKGROUND: Previous reports have suggested that one-fourth of amblyopic subjects present with recurrence of amblyopia even after successful primary treatment. Risk factors for amblyopia recurrence are insufficiently studied, but therapy weaning has recently been suggested as a method for reducing the risk of recurrence. METHODS: In the second phase of a prospective study examining the results of treatment for amblyopia, all 35 children with successful primary amblyopia treatment were put on maintenance therapy. Maintenance therapy consisted of low-intensity patching, atropine, or blurring filter, and all children were regularly examined up to at least 8 years of age. Mean age at start of maintenance therapy was 4.3 years (range, 2-7 years). RESULTS: Of the 35 cases with successful primary treatment, 6 cases deteriorated >or=0.2 logMAR, which was considered recurrence of amblyopia. Two of these cases had a second recurrence. Notably, all six subjects presenting with recurrence had microstrabismus. This finding was just outside statistical significance at the 95% confidence level (p = 0.06), but the sample size was small. All but one of the recurrences appeared within 6 months after successful primary treatment. Age at successful primary treatment or initial interocular difference of visual acuity did not affect the risk of recurrence. CONCLUSIONS: Recurrence of amblyopia occurred in 17% of patients despite maintenance therapy and was associated with microstrabismus. The majority of recurrences occurred within the first 6 months after primary treatment. PMID- 17419082 TI - Stereoacuity and binocular visual acuity in prism-induced exodeviation. AB - BACKGROUND: Intermittent exotropia may be controlled by accommodative or motor convergence. Previous studies have reported that reduced binocular visual acuity in intermittent exotropia is due to the accommodation required to control the deviation. To test this hypothesis, we simulated convergence stress by inducing exodeviations in normal volunteers to investigate whether the transition from nonfused to fused is associated with a gradual or immediate transition in stereoacuity and binocular visual acuity. METHODS: Convergence stress was induced in 25 visually normal adults with 40(Delta) base-out prism and reduced stepwise in increments of 5(Delta) until 20(Delta), and 2(Delta) thereafter. Stereoacuity (Frisby Davis Distance [FD2] and the Distance Randot [DR]) and binocular visual acuity were measured at each step. For each subject, the recovery of fine stereoacuity (< or =60 arcsec) was categorized as immediate (nil to fine) or gradual (nil to moderate to fine). RESULTS: Twenty-four of 25 (96%) showed gradual recovery of fine stereoacuity on either FD2 or DR. Median binocular visual acuity at baseline, first level of fusion, and subsequent levels was 20/15. CONCLUSIONS: Under convergence stress, the transition from nonfused to fused is accompanied by a gradual recovery of fine stereoacuity in most individuals, consistent with some studies of patients with intermittent exotropia. Nevertheless, this degradation of stereoacuity was not associated with decreased binocular visual acuity, suggesting that accommodative convergence may not be recruited to restore and maintain binocularity under conditions of convergence stress. PMID- 17419083 TI - One-muscle surgery in small-angle residual esotropia. AB - PURPOSE: To report the outcome of unilateral lateral rectus resection for treatment of small-angle residual esotropia following bilateral medial rectus muscle recession. METHODS: A retrospective medical record review was performed for all patients who had undergone bilateral medial rectus muscle recession for congenital esotropia prior to 6 years of age that required further surgical treatment of residual esotropia. We compared two different dosing strategies for resection of a single lateral rectus muscle in the nondominant eye. In group 1, the amount of resection was calculated by doubling the angle of strabismus and applying the recommended surgical dosage to one lateral rectus muscle. In group 2, the amount of unilateral resection was the same as the bilateral dosage for the measured angle, but augmented by 1.5 mm. Postoperative evaluation was performed 1 and 6 months after surgery. RESULTS: Data from 35 patients were analyzed, 17 in group 1 and 18 in group 2. No significant intergroup difference was noted in terms of age at first surgery (p = 0.266), initial surgical dosage (p = 0.693), residual angle of esotropia (p = 0.881), or age at reoperation (p = 0.679). Postoperative alignment was better in group 1 patients at 6 months than at 1 month (residual deviation 3.5(Delta) versus 6.7(Delta), p = 0.022). CONCLUSIONS: Resection of a single lateral rectus muscle with the surgical dosage calculated by doubling the angle of strabismus and applying the recommended surgical dosage to one lateral rectus muscle is a treatment option for patients with small-angle residual esotropia following bilateral medial rectus muscle recession. PMID- 17419084 TI - Cytokines, bone turnover markers and weight change in candidates for lung transplantation. AB - Weight loss in chronic obstructive pulmonary disease (COPD) is associated with increased morbidity and may negatively affect bone mineral density. Increased serum levels of cytokines such as tumour necrosis factor (TNF)-alpha have been associated with weight loss and with bone resorption. We studied the association between systemic inflammation, markers for bone turnover and recent weight change in underweight (n=48) and normal-weight patients (n=23) candidates for lung transplantation where the majority (56%) had COPD. Osteoporosis or osteopenia was present in all the diagnostic groups. The resulting model of linear regression in COPD patients showed that for the 1-CTP (a marker of bone resorption) model, the total variation of 61% was explained by recent weight change, sTNF-alpha receptor(R)II, dose of prednisolon and age. The resulting model of linear regression in the whole group of patients showed that the total variation of 72% was explained by recent weight change, sTNF-alpha RI, diagnosis (COPD/other diagnosis), dose of prednisolon and C-reactive protein. In conclusion, our results showed that serum concentration of 1-CTP was positively associated with sTNF-alpha receptor II and negatively with recent weight change in patients with advanced COPD. Recent weight loss in both the underweight and normal-weight patients showed to be a more important contributor than recent weight loss only in underweight patients for explaining variations in 1-CTP. PMID- 17419085 TI - Ultrasonic irradiation accelerated cyclopalladated ferrocenylimines catalyzed Suzuki reaction in neat water. AB - Both conventional heating and ultrasound effect on the cyclopalladated ferrocenylimines catalyzed Suzuki reaction of phenylboronic acid with a range of arylhalides in neat water was investigated. Heterogenous reaction of electron withdrawing arylchlorides with phenylboronic acid could also result in good yields by using Cat. 2. It was found that the ultrasonic irradiation could dramatically accelerate the Suzuki reaction to achieve comparable results. PMID- 17419086 TI - A statistical thermodynamic approach to sonochemical reactions. AB - The calculation of the equilibrium constants K of the sonolysis reactions of CO2 into CO and O atom, the recombination of O atoms into O2 and the formation of H2O starting with H and O atoms, has been studied by means of statistical thermodynamic. The constants have been calculated at 300 kHz versus the pressure and the temperature according to the extreme conditions expected in a cavitation bubble, e.g. in the range from ambient temperature to 15200 K and from ambient pressure to 300 bar. The decomposition of CO2 appears to be thermodynamically favored at 15200 K and 1 bar with a constant K1=1.52 x 10(6), whereas the formation of O2 is not expected to occur (K2=1.8 x10(-8) maximum value at 15200 K and 300 bar) in comparison to the formation of water (K3=3.4 x 10(47) at 298 K and 300 bar). The most thermodynamic favorable location of each reactions is then proposed, the surrounding shell region for the thermic decomposition of CO2 and the wall of the cavitation bubble for the formation of water. Starting from a work of Henglein on the sonolysis of CO2 in water at 300 kHz, the experimental amount of CO formed (7.2 x 10(20)molecules L(-1)) is compared to the theoretical CO amount (1.4 x 10(27)molecules L(-1)) which can be produced by the sonolysis of the same starting amount CO2. With the help of the literature data, the number of cavitation bubble has been evaluated to 6.2 x 10(15) bubbles L(-1) at 300 kHz, in 15 min. This means that about 1 bubble on 1900000 is efficient for undergoing the sonolysis of CO2. PMID- 17419087 TI - An efficient and convenient procedure for the synthesis of 2-alkyl-2-alkoxy-1,2 di(furan-2-yl)ethanone under ultrasound in the presence of solid-liquid phase transfer catalysis conditions. AB - Irradiation with ultrasound is found to facilitate the synthesis of 2-alkyl-2 alkoxy-1,2-di(furan-2-yl)ethanone in the presence of tetrabutylammonium bromide (TBAB) as solid-liquid phase transfer catalysis. A higher yields (>70%) and shorter reaction time (<15 min) give this method obvious advantages over the conventional ones. PMID- 17419088 TI - Clinical validation of three-dimensional tortuosity metrics based on the minimum curvature of approximating polynomial splines. AB - The clinical recognition of abnormal vascular tortuosity is important in the diagnosis of many diseases. Metrics based on three-dimensional (3D) curvature, using approximating polynomial spline-fitting to "data balls" centered along the mid-line of the vessel, minimize digitization errors and give tortuosity values largely independent of the resolution of the imaging system. We applied two of these metrics to a number of clinical vascular systems, using both 2D and 3D datasets. Using abdominal aortograms of low tortuosity, we established their validity by their strong correlation with the ranking of an expert panel of three vascular surgeons. The values of the Spearman rank correlation coefficient between our rankings, using a data ball radius of one-quarter of the local vessel radius, and the average ranking of the expert panel were 0.96 (with a 95% confidence interval of [0.91, 0.99]) for the mean curvature and 0.98 ([0.94, 0.99]) for the root-mean-square (RMS) curvature. These confidence intervals indicate that our automated analysis is producing rankings whose reliability is similar to that of a human expert, and is significantly better than that achieved with existing algorithms. The metrics provided good discrimination between vessels of different tortuosity for both 2D and 3D datasets, and produced values sufficiently discriminating to assess the relative utility of arteries for endoluminal repair of aneurysms. PMID- 17419089 TI - Ezrin/radixin/moesin: versatile controllers of signaling molecules and of the cortical cytoskeleton. AB - Ezrin, radixin and moesin (ERM) proteins are widely distributed proteins located in the cellular cortex, in microvilli and adherens junctions. They feature an N terminal membrane binding domain linked by an alpha-helical domain to the C terminal actin-binding domain. In the dormant state, binding sites in the N terminal domain are masked by interactions with the C-terminal region. The alpha helical domain also contributes to masking of binding sites. A specific sequence of signaling events results in dissociation of these intramolecular interactions resulting in ERM activation. ERM molecules have been implicated in mediating actin-membrane linkage and in regulating signaling molecules. They are involved in cell membrane organization, cell migration, phagocytosis and apoptosis, and may also play cell-specific roles in tumor progression. Their precise involvement in these processes has yet to be elucidated. PMID- 17419090 TI - Genotoxic effects in wild rodents (Rattus rattus and Mus musculus) in an open coal mining area. AB - Coal is a mixture of a variety of compounds containing mutagenic and carcinogenic polycyclic aromatic hydrocarbons. Exposure to coal is considered as an important non-cellular and cellular source of reactive oxygen species that can induce DNA damage. In addition, spontaneous combustion can occur in coal mining areas, further releasing compounds with detrimental effects on the environment. In this study the comet assay was used to investigate potential genotoxic effects of coal mining activities in peripheral blood cells of the wild rodents Rattus rattus and Mus musculus. The study was conducted in a coal mining area of the Municipio de Puerto Libertador, South West of the Departamento de Cordoba, Colombia. Animals from two areas in the coal mining zone and a control area located in the Municipio de Lorica were investigated. The results showed evidence that exposure to coal results in elevated primary DNA lesions in blood cells of rodents. Three different parameters for DNA damage were assessed, namely, DNA damage index, migration length and percentage damaged cells. All parameters showed statistically significantly higher values in mice and rats from the coal mining area in comparison to the animals from the control area. The parameter "DNA Damage Index" was found to be most sensitive and to best indicate a genotoxic hazard. Both species investigated were shown to be sensitive indicators of environmental genotoxicity caused by coal mining activities. In summary, our study constitutes the first investigation of potential genotoxic effects of open coal mining carried out in Puerto Libertador. The investigations provide a guide for measures to evaluate genotoxic hazards, thereby contributing to the development of appropriate measures and regulations for more careful operations during coal mining. PMID- 17419091 TI - Sporadic colorectal cancer and individual susceptibility: a review of the association studies investigating the role of DNA repair genetic polymorphisms. AB - Mutations in one of the DNA repair genes are one of the most common reasons for cancer, and it may be assumed that the individual genetic background modulating the DNA repair capacity may affect the susceptibility to cancer. Numerous polymorphisms (mainly SNPs) have been identified for DNA repair genes, although their functional outcome and phenotypic effect is often unknown. The aim of the present review is to evaluate the studies investigating a possible influence of DNA repair polymorphisms in the risk of sporadic colorectal cancer and/or adenoma. Overall, no relevant common findings emerge among the studies, except for some statistically significant associations between polymorphisms in the XRCC1 and XPD genes, mainly for colorectal adenoma risk. Other individual associations remain to be confirmed. This inconclusive data may suggest that the modulation of cancer risk depends not only on a single gene/SNP, but also on a joint effect of multiple polymorphisms (or haplotypes) within different genes or pathways, in close interaction with environmental factors. The relevance of many low-penetrance genes in cancer susceptibility is supposed to be very subtle. Several reviewed association studies revealed weaknesses in their design. However, there has been a progressive improvement over the years in aspects such as simultaneous genotyping and combined analyses of different polymorphisms in larger numbers of patients and controls, as well as stratification of results by ethnicity, gender, and tumor localization. This gained experience shows that only carefully designed studies of a sufficient statistical power may resolve the relationships between polymorphisms and colorectal cancer risk. PMID- 17419092 TI - Optical absorption spectroscopic studies on holmium(III) complexes with beta diketone and heterocyclic amines: the environment effect on 4f-4f hypersensitive transitions. AB - The optical absorption spectra of [Ho(acac)(3)(H(2)O)(2)].H(2)O, [Ho(acac)(3)phen] and [Ho(acac)(3)bpy] (where acac is the anion of acetylacetone; phen is 1,10-phenanthroline and bpy is 2,2'-bipyridyl) complexes in the visible region, in a series of non-aqueous solvents (methanol, ethanol, isopropanol, chloroform, acetonitrile and pyridine.) have been analyzed. The largest intensity variation was observed in the (5)G(6)<--(5)I(8) (centered at 450 nm), and (5)G(5), (3)H(5), (3)H(6)<--(5)I(8) (centered at 360 nm) transition regions. The band shape and oscillator strength of the hypersensitive transitions display pronounced changes as compared to Ho(3+) aqua-ion. The band shapes of the hypersensitive transitions show remarkable changes on passing from aqueous solution to various non-aqueous solutions which is the result of change in the environment about the Ho(III) ion in the various solutions and suggests coordination of solvent molecule(s), in some cases. The results clearly show that among the solvents studied pyridine is the most effective in promoting the 4f-4f spectral intensity. It has been inferred from this study that 2,2'-bipyridyl is a stronger ligand for heavier lanthanides. A comparative account of hypersensitivity in the present complexes with those of other adducts of Ho(beta diketoenolate)(3) with heterocyclic amines is discussed. The TGA analyses showed that the phen complex is thermally more stable over its bpy analogue. PMID- 17419093 TI - Synthesis and spectroscopic properties of lanthanide nitrate complexes with a new amide-based quinoxaline-2,3-dione ligand. AB - Solid complexes of lanthanide nitrate with 1,4-di(N,N-di-n-butyl-acetamido) quinoxaline-2,3-dione (L), [Ln(N03)3L.H2O] (Ln=La, Nd, Eu, Gd, Tb, Er), have been prepared and characterized by elemental analysis, IR, UV-vis spectra and conductivity measurements. The fluorescence property of the europium complex in solid state and in MeCN, acetone, AcOEt and THF was studied. Under the excitation, the europium complex exhibited characteristic emissions of europium. The result indicates that the triplet state energy level of the ligand matches better to the resonance level of Eu(III) than Tb(III) ion. PMID- 17419094 TI - Reversal in solvatochromism in some novel styrylpyridinium dyes having a hydrophobic cleft. AB - The influence of solvent polarity on the electronic transition of four different N-hexadecyl styrylpyridinium dyes has been investigated in 15 solvents. The E(T)(30) scale has been used to propose a quantitative approach towards the relative stability of the electronic ground and excited state species. The extents of contribution of dipolar aprotic solvents towards the solvation of the excited species have been determined to be 42-48% for some of the dyes. Instead of a steady solvatochromism, all the dyes suffer a reversal in solvatochromism. The transitions of the solvatochromism, referred to as solvatochromic switches, are found to be at E(T)(30) values of approximately 50 for methyl and N,N dimethylamino substituted dyes while at 37.6 for hydroxyl substituted dye and approximately 45 for 4-(1-methyl-2-phenylethenyl) pyridinium dye. A reversal in the trend of solvent effect in the later dye corresponding to 4-(4-methyl styryl)pyridinium dye has been attributed to an analogy of series and parallel electron flow. PMID- 17419095 TI - Modificomics: posttranslational modifications beyond protein phosphorylation and glycosylation. AB - Posttranslational modifications of proteins possess key functions in the regulation of various cellular processes. While they facilitate fast, location specific and transient reactions to changing conditions in the first place they enhance the already high complexity of a cellular proteome by orders of magnitude. Furthermore, they can utterly alter the properties of the modified protein, thus making a timely analysis even more difficult. While several standardized methods for the analysis of protein phosphorylation and glycosylation have been established most other modifications require tailor-made solutions for a comprehensive analysis. Therefore, we will provide guidelines for the analysis of some important posttranslational modifications that are underrepresented in contemporary literature. PMID- 17419096 TI - Left ventricular reconstruction by modified linear technique with absorbable suture. AB - BACKGROUND: Linear and endoventricular techniques of left ventricular (LV) reconstruction often utilise prosthetic material for epicardial reinforcement or endocardial patch. We report a technique of LV aneurysm repair using absorbable suture without prosthetic material. METHODS: Between November 1999 and August 2004, 55 patients underwent linear LV reconstruction, for ischaemic cardiomyopathy, using only continuous 3/0 polydioxanone suture. Survival, functional class and echocardiographic outcomes are reported. RESULTS: Fifty-two patients (mean age 64+/-10 years) were studied. There was no perioperative mortality and the Kaplan-Meier survival at five years was 81%. Fifty-one patients underwent postoperative follow up (mean 28 months) with echocardiography (mean 20 months). There were no recurrent aneurysms. The postoperative LV eccentricity index (EI) was 0.72 (CI 0.48-0.9). Mean LV ejection fraction increased from 0.33+/-0.09 preoperatively to 0.41+/-0.15 at late follow up (mean within patient change 0.08+/-0.15, p=0.003). Mean symptom class improved from 3.3+/-0.8 to 1.6+/ 0.7 (mean within patient improvement 1.6+/-1.0, p<0.001). Mitral regurgitation of grade 2/4 was identified in six patients and grade 3/4 in one patient. CONCLUSION: Linear repair with absorbable suture material and without prosthetic material may be safely undertaken with good early and mid-term results. PMID- 17419097 TI - Contemporary treatment of in-stent restenosis and the incidence of recurrent in stent restenosis in the era of drug-eluting stents. AB - BACKGROUND: Optimal treatment of in-stent restenosis (ISR) remains uncertain in the era of drug-eluting stents (DES). This study aims to determine contemporary treatment of ISR and to assess recurrent ISR rates in the era of DES. METHODS: We examined 60 patients presenting for treatment of ISR (one lesion per patient) who were enrolled in the Melbourne Interventional Group Registry (4% of total population of 1423 patients) between April 2004 and January 2005. Twelve-month follow-up is complete for all patients. RESULTS: The majority of ISR treated occurred in bare metal stents [BMS (n=52, 87%)] and had a focal (<10 mm) pattern of ISR (53%). In-stent restenosis of DES occurred in eight (13%) patients. The majority of ISR were treated with additional stenting with a preference for DES over BMS in almost all cases. At 12 months, one patient died of non-cardiac cause and four patients (7%) presented with recurrent ISR. The incidence of recurrent ISR in DES was 5% (n=3). No late thrombosis was reported despite only 50% of patients having >or=12 months of clopidogrel therapy. CONCLUSIONS: Our study suggests drug-eluting stents are safe, effective and the preferred therapy for in stent restenosis. The incidence of recurrent drug-eluting stent restenosis at 12 months is low. PMID- 17419098 TI - Coronary artery aneurysms associated with a Paclitaxel coated stent. AB - Coronary artery aneurysm associated with the Paclitaxel drug eluting stent (Taxus) is a rare complication. We describe the case of a 71-year-old female, who developed two coronary artery aneurysms in her right coronary artery associated with the insertion of a Taxus stent. Although no adverse clinical outcome resulted, this case highlights a potential problem going forward. PMID- 17419099 TI - Spontaneous subscapular abscess: a rare surgical condition. AB - Subscapular abscess is a rare clinical problem. Herein we describe a young male patient who presented with a subscapular abscess and was managed successfully with surgical drainage and debridement of the cavity. PMID- 17419100 TI - Bleeding events with antithrombotic therapy in patients with unstable angina or non-ST-segment elevation myocardial infarction; insights from a large clinical practice registry (GRACE). AB - BACKGROUND: Thienopyridine use, in particular clopidogrel in acute coronary syndromes, has been associated with an improvement in outcome. However, little information is available regarding their bleeding risk when used in combination with other antithrombotic agents and revascularisation. METHODS: In a large, multinational, prospective registry, the Global Registry of Acute Coronary Events, the major bleeding rate (using GRACE criteria) of 27,358 patients with unstable angina or non-ST-elevation myocardial infarction was recorded during index admission. The interaction of thienopyridines on major bleeding with other antithrombotic agents and with revascularisation was analysed. RESULTS: The 11,478 patients who received thienopyridines during hospitalisation experienced a significant increase in major bleeding (2.8% with thienopyridines, 2.2% without thienopyridines; p=0.002). No significant interaction with glycoprotein IIb/IIIa inhibitors and thienopyridines was seen with regard to bleeding. Thienopyridines with unfractionated heparin did not alter bleeding risk, but thienopyridines with low molecular weight heparin was associated with a significant excess of bleeding (2.1% with thienopyridines, 1.3% without thienopyridines; p=0.004). There was no significant difference in major bleeding with thienopyridines in patients who did or did not undergo revascularisation. CONCLUSIONS: Major bleeding was increased in patients receiving thienopyridines. No increase in bleeding risk was seen in patients having revascularisation. PMID- 17419101 TI - [Gastrointestinal tuberculosis as the main manifestation of systematic tuberculosis]. PMID- 17419103 TI - Readers' and author's responses to "medicine and motherhood". PMID- 17419102 TI - Readers' and author's responses to "medicine and motherhood". PMID- 17419104 TI - Readers' and author's responses to "body contouring after bariatric surgery is critical for optimal cosmetic results". PMID- 17419105 TI - Readers and authors respond to "introducing the doctor of nursing practice". PMID- 17419106 TI - Readers and authors respond to "introducing the doctor of nursing practice". PMID- 17419107 TI - Readers' and author's responses to "information on complementary and alternative medicine in US government databases is biased". PMID- 17419108 TI - Readers' and author's responses to "information on complementary and alternative medicine in US government databases is biased". PMID- 17419109 TI - Readers' and author's responses to "information on complementary and alternative medicine in US government databases is biased". PMID- 17419110 TI - Readers' and author's responses to "information on complementary and alternative medicine in US government databases is biased". PMID- 17419111 TI - Readers respond to "resting metabolic rate: a critical, primary care screening test". PMID- 17419112 TI - Readers respond to "resting metabolic rate: a critical, primary care screening test". PMID- 17419113 TI - Readers respond to "resting metabolic rate: a critical, primary care screening test". PMID- 17419114 TI - Readers respond to "the $1.6 trillion question: if we're spending so much on healthcare, why so little improvement in quality?". PMID- 17419115 TI - Readers' and author's responses to "fixing the US physician shortage requires many more slots for resident physicians in training". PMID- 17419116 TI - Readers' and author's responses to "fixing the US physician shortage requires many more slots for resident physicians in training". PMID- 17419117 TI - Readers' and author's responses to "fixing the US physician shortage requires many more slots for resident physicians in training". PMID- 17419118 TI - Readers' and author's responses to "fixing the US physician shortage requires many more slots for resident physicians in training". PMID- 17419119 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17419120 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17419121 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17419122 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17419123 TI - Readers' responses and author's reply to "the 'diabesity' epidemic: let's rehabilitate America". PMID- 17419124 TI - [Functional food for the skin]. PMID- 17419125 TI - Calls to curb deep-ocean trawling. PMID- 17419127 TI - Pressure off. PMID- 17419126 TI - Seeking a sea change. PMID- 17419128 TI - Loren Riesberg. PMID- 17419129 TI - [Reiter disease or reactive arthritis?]. AB - There is an ongoing international discussion on whether the condition reactive arthritis should be named after a former Nazi functionary. The German dermatological community should participate in this debate. In 1916, Hans Reiter described a disease with the symptoms urethritis, conjunctivitis, and arthritis, which was later named after him. After becoming titular professor in May 1918, Reiter was appointed director of the regional public health department Mecklenburg-Schwerin in 1926. At the same time he taught social hygiene at the University of Rostock, where he was appointed full professor in 1928. In 1931, Hans Reiter became a member of the National Socialist German Workers Party (NSDAP). In July 1932 he was elected representative of the NSDAP to the seventh assembly of Mecklenburg-Schwerin. After becoming its acting director in July 1933, Reiter was appointed president of the Reich public health department in Berlin on October 1, 1933. Both his excellent professional qualifications, as well as his National Socialist attitudes, were considered key criteria for taking over this important position. As the president of the Reich public health department, Reiter was said to have known about the conduct of experiments with typhus-fever at the concentration camp Buchenwald in which 250 humans died. From the end of the Second World War until 1947, Reiter was imprisoned in the Nuremberg Prison for War Criminals, but never convicted of a crime. PMID- 17419130 TI - [15th Consensus Conference on Anti-infective Therapy]. PMID- 17419131 TI - Correction: Foreword for critical access hospital manual. PMID- 17419132 TI - Correction: Top standards compliance issues for 2006, behavioral health care and office-based surgery. PMID- 17419133 TI - Beyond electrostatics. PMID- 17419134 TI - A sound idea. PMID- 17419136 TI - A sound idea. PMID- 17419135 TI - Beyond electrostatics. PMID- 17419137 TI - Pakistan struggles to eradicate polio. PMID- 17419138 TI - Herpes treatment may limit HIV transmission and progression. PMID- 17419139 TI - USA increases funds to HIV/AIDS and tuberculosis programmes. PMID- 17419140 TI - Mind in general. PMID- 17419141 TI - [Tying and untying the knot: women's networks and separations in Rome, 17th-18th centuries]. PMID- 17419142 TI - [Dress, gender, and citizenship in modern Tuscany in the 16th-17th centuries]. PMID- 17419143 TI - [An "old" source for new problems: censuses and the study of preindustrial mobility]. PMID- 17419144 TI - ["Perfect woman or hermaphrodite?" Physiology and "gender" in an 18th-century monastery]. PMID- 17419145 TI - [The theater of justice: guilty women and public opinion in liberal Italy]. PMID- 17419146 TI - FDA approves Avastin in combination with chemotherapy for first-line treatment of most common type of lung cancer. PMID- 17419147 TI - Cancer helps itself by sending out chemicals that cripple nearby lymph nodes, making them unable to resist cancer spread. PMID- 17419148 TI - Cancer stem cells linked to radiation resistance. PMID- 17419149 TI - New Year's resolution: become an advanced practice nurse. Interview by Barbara Zeiger. PMID- 17419150 TI - Beyond imatinib: second generation c-KIT inhibitors for the management of gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal cancer of teh gastrointestinal tract. They are characterized by the expression of KIT. Therapeutically, metastatic GISTs are effectively treated by imatinib, a tyrosine kinase inhibitor (TKI) with activity against KIT and platelet-derived growth factor receptor. Gastrointestinal stromal tumors refractory to standard therapy with imatinib are a clinical challenge. This has lead to the clinical testing of a variety of agents used alone or in combination with other TKIs. Sunitinib, a multitargeted TKI, is the first drug available fort eh treatment of these patients. Additional trials are ongoing, evaluating the efficacy of the novel KIT TKIs AMG 706 and AMN 107 (nilotinib). RAD001, PKC412, and bavacizumab are being tested in conjunction with imatinib. Lastly, the heat-shock protein-90 inhibitor IPI-540 is also in phase I evaluation in imatinib-refractory patients with GIST. The future management of GIST is likely to be altered by the availability of more agents and by better biologic understanding of the patient populations each agent best treats. PMID- 17419151 TI - Administrative costs for advance payment of health coverage tax credits: an initial analysis. AB - Health Coverage Tax Credits (HCTCs), created under the Trade Act of 2002, pay 65 percent of health insurance premiums for certain workers displaced by international trade and early retirees. These credits can be paid directly to insurers when monthly premiums are due, in advance of annual tax return filing. While HCTC administrative costs have fallen significantly since program start ups, they still comprise approximately 34 percent of total spending. Changes to the HCTC program could lower administrative costs, but the size of the resulting savings is unknown. These findings have important implications for any future tax credit plan intended to cover the uninsured. PMID- 17419152 TI - Development of fatigue and discomfort in the upper trapezius muscle during light manual work. AB - Optimization of the temporal aspects of task design requires a better understanding of the development of muscle fatigue in the neck and shoulder region over time. The objective of the study was to investigate this in two production companies and to determine the relationship between objective and subjective estimates of fatigue. Indicators of fatigue were obtained through electromyography (EMG) during test contractions and ratings of perceived discomfort. EMG amplitude increased during the day in both case studies while mean power frequency decreased only in one case. In both cases, a more detailed frequency analysis of the EMG signals showed an increase in lower frequency power accompanied by a decrease in higher frequency power. Local perceived discomfort in the neck and shoulder increased over the course of the day in both cases. However, no clear relationship between perceived discomfort and objective indicators of fatigue was found. Obtaining sufficient sensitivity to detect effects of temporal aspects of task design probably requires complementary or more refined methods (e.g. EMG arrays, mechanomyography). PMID- 17419153 TI - The effects of pause software on the temporal characteristics of computer use. AB - The study investigated the natural work-pause pattern of computer users and the possible effects of imposing pause regimes on this pattern. Hereto, the precise timing of computer events was recorded across a large number of days. It was found that the distribution of the pause durations was extremely skewed and that pauses with twice the duration are twice less likely to occur. The effects of imposing pause regimes were studied by performing a simulation of commercially available pause software. It was found that depending on the duration of the introduced pause, the software added 25-57% of the pauses taken naturally. Analysis of the timing of the introduced pauses revealed that a large number of spontaneous pauses were taken close to the inserted pause. Considering the disappointing results of studies investigating the effects of introducing (active) pauses during computer work, this study has cast doubt on the usefulness of introducing short duration pauses. PMID- 17419154 TI - Postural implications of obtaining line-of-sight for seated operators of underground mining load-haul-dump vehicles. AB - Operators of load-haul-dump (LHD) vehicles use awkward postures that may be held statically and at extreme ranges of motion for long shift periods to spot hazards in underground mining. This study examined postural variables associated with three amounts of seat rotation intended to maximize line-of-sight during forward driving. Three different models, representing the 1st, 50th and 99th percentile male for height and weight, were positioned with appropriate hand and foot constraints in the virtual LHD cab modelled in Classic JACK v4.0. A total of 15 virtual movement strategies were developed to model the postural behaviour of typical workers and each virtual subject was tested, first with the seat in a neutral 0 degrees position and then with it rotated counter-clockwise to 20 degrees and 45 degrees . Results revealed that reductions in trunk rotation, trunk lateral bend and neck rotation were associated with the seat rotation intervention. The general relationship observed was that as seat rotation increased, view of critical visual attention locations and visible line-of-sight area increased while postural load variables decreased. For the most part, 20 degres of seat rotation was beneficial but 45 dgrees produced significantly greater changes to postural load and visible visual attention locations. PMID- 17419155 TI - A new input device: comparison to three commercially available mouses. AB - This study was conducted to simultaneously compare the postural demands and performance of a new human-centred computer input device to three devices currently on the market. It was hypothesized that the new device would perform as well as the commercial devices while requiring less postural strain. A total of 24 experienced computer users performed a series of modified Steering and Fitts' Law Tests while their postural behaviour was captured using an opto-electric system. Analysis of the postural data revealed strong similarities between the new device and the commercially available devices, including some similarities that are not suggested in the literature. Analysis of the performance data reveals no significant difference between the new device and most commercial devices and suggests further examination of the difference between familiarity and mastery. This study has shown that it is possible to use the new device in a relaxed posture and yet achieve the same accuracy as commercial devices at no more postural risk than when the traditional mouse is used at a customized, ergonomic workstation. PMID- 17419156 TI - A quantitative meta-analytic examination of whole-body vibration effects on human performance. AB - Whole-body vibration exerts a substantive influence in many work environments. The primary objective for this work was to quantify such effects by identifying those moderating variables that influencethe degree to which performance is affected. To achieve this, a comprehensive meta-analysis was conducted, which synthesized the existing research evidence, A total of 224 papers and reports were identified and, from these 115 effect sizes were derived from 13 experiments that survived the screening procedure. Results indicate that vibration acts to degrade the majority of goal-related activities, especially those with high demands on visual perception and fine motor control. Gaps in the current research literature are dentified and suggestions offered with regard to a more theoretically-driven approach to testing vibration effects on human performance. PMID- 17419157 TI - Comparison of anaesthetists' activity patterns in the operating room and during simulation. AB - This study investigated the behavioural aspects of ecological validity of anaesthesia simulation environments using a task analysis approach. Six anaesthesists were observed during two cases performed in the operating room (OR), one routine and two critical incident simulation scenarios. A two-way MANOVA for repeated measures was performed with the independent variables Case (OR/SIM-R/SIM-CI) and Phase Induction/ Maintenance (Emergence), the latter being a repeated measure. Dependent variables were the proportion of each phase spent on each of the observation categories. Statistically significant main effects for Phase concerning communication, monitoring, manual tasks and documentation, for Case concerning communication and documentation, and a significant interaction effect for Phase x Case concerning manual tasks and other were found. Increased action density (i.e. amount of co-occurring activities) was observed during Induction, Emergence and the Management of simulated critical events. The similarities and differences in anaesthetists' activity patterns identified in this study will help to further improve the ecological validity of simulation environments as research settings. PMID- 17419158 TI - Effects of psychosocial and individual factors on physiological risk factors for upper extremity musculoskeletal disorders while typing. AB - Psychosocial factors are hypothesized to contribute to work-related musculoskeletal disorder (WMSD) development, although previous research has been largely epidemiological or has focused primarily on the shoulders, back and neck. The objective of this study was to quantify the effects of mental workload and time pressure on perceived workload and physiological responses of the distal upper extremity. A total of 18 typists completed nine 5-min typing sessions representing three levels of time pressure and mental workload. Levels were manipulated by adjusting typing speed and by requiring participants to perform arithmetic tasks while typing. Outcomes were measured in muscle activation levels, wrist postures and movements, key strike force and subjective assessments of workload. In general, increased time pressure increased muscle activation, key strike force and wrist deviations; and increased mental workload increased key strike force. Mental workload and time pressure mediated physical risk factors during typing to increase WMSD risk for the distal upper extremity. PMID- 17419159 TI - Mild body cooling impairs attention via distraction from skin cooling. AB - Many contemporary workers are routinely exposed to mild cold stress, which may compromise mental function and lead to accidents. A study investigated the effect of mild body cooling of 1.0 degree C rectal temperature (Tre) on vigilance (i.e. sustained attention) and the orienting of spatial attention (i.e. spatially selective processing of visual information). Vigilance and spatial attention tests were administered to 14 healthy males and six females at four stages (pre immersion, deltaTre = 0, -0.5 and - 1.0 degree C ) of a gradual, head-out immersion cooling session (18-25 deltaC water), and in four time-matched stages of a contrast session, in which participants sat in an empty tub and no cooling took place. In the spatial attention test, target discrimination times were similar for all stages of the contrast session, but increased significantly in the cooling phase upon immersion (deltaTre = 0 degrees C), with no further increases at deltaTre = -0.5 and - 1.0 degree C. Despite global response slowing, cooling did not affect the normal pattern of spatial orienting. In the vigilance test, the variability of detection time was adversely affected in the cooling but not the contrast trials: variability increased at immersion but did not increase further with additional cooling. These findings suggest that attentional impairments are more closely linked to the distracting effects of cold skin temperature than decreases in body core temperature. PMID- 17419160 TI - Visual lobe shape and search performance for targets of different difficulty. AB - In quantitative models of visual search it has usually been assumed that visual lobe area shape was sufficiently regular to be approximated by a circle or ellipse. However, the irregularities in visual lobe shapes that have been found in studies involving extensive lobe mapping have suggested that lobe shape may have important implications for visual search performance and for the accuracy of mathematical models used for performance prediction. However, no systematic research on the relationship between the shape aspect of visual lobes and search performance seems to have been carried out and no comparisons of visual lobe shape characteristics under the effect of target difficulty have been reported. The current study was conducted to achieve two major objectives in two experiments. Experiment 1 used two different targets (letter 'O' and letter 'Y') to map the visual lobes of subjects in order to provide a systematic and quantitative comparison of lobe shape characteristics and experiment 2 was to investigate the correlation of visual lobe shape characteristics with visual search time under the effect of target difficulty. The visual lobes of 28 subjects were mapped on 24 imaginary and regularly spaced meridians originating from the centre of the visual field to resemble the full field mapping situation. Five categories of shape indices, viz. roundness, boundary smoothness, symmetry, elongation and shape regularity were investigated. The results of this study demonstrated that the visual lobe shapes of subjects elongate horizontally with medium level of roundness, high levels of boundary smoothness, symmetry and regularity for an easy target (O) against a homogeneous background of 'X's. When a difficult target (Y) was used, the visual lobes of the subjects were still elongated horizontally but to a smaller extent and with a low level of roundness, medium level of boundary smoothness and regularity and a similar high level of symmetry to the easy target. Moreover, significant correlations between shape indices and visual search time were found, suggesting mathematical models for predicting search time should not merely rely on area but also should consider visual lobe shape indices. Finally, a universal mathematical model containing several visual lobe shape indices was developed, which was applicable in the prediction of visual search time for a range of similar search tasks. PMID- 17419162 TI - Medical licensure and state regulation of medical practice. PMID- 17419161 TI - From my perspective. PMID- 17419163 TI - The future of sugical simulation and surgical robotics. PMID- 17419164 TI - IOM and ACS warn of the impending crisis in emergency care: emergency departments overwhelmed, underfunded, and dangerously fragmented. PMID- 17419165 TI - Using medical malpractice closed claims data to reduce surgical risk and improve patient safety. PMID- 17419166 TI - Surgeon melds music and medicine. PMID- 17419167 TI - Program for Accreditation of Education Institutes continues to grow. PMID- 17419168 TI - My girdle is killing me. PMID- 17419169 TI - [Pancreaticopleural fistula]. PMID- 17419170 TI - [Surgical ablation for atrial fibrillation: the future of cardiac surgery?]. PMID- 17419171 TI - [Residual echocardiographic findings and NT-proBNP in asymptomatic adult patients after radical correction of Fallot's tetralogy]. AB - OBJECTIVE: Define the profile of NT-proBNP values in asymptomatic adult patients after radical correction of Tetralogy of Fallot (TOF) and identify links between possible increase in NT-proBNP and residual echocardiographic findings. METHODOLOGY: NT-proBNP samples were taken from and a detailed echocardiographic examination was performed in 21 adult stabilised patients after radical correction of TOF in childhood. The results were submitted for statistical analysis. RESULTS: The incidence of low values of the S wave < 11.5 cm/s of tricuspid anulus evaluated by tissue Doppler echocardiography (TDI) (P < 0.05) was significantly higher in patients with NT-proBNP > 125 pg/ml. All patients with impaired right ventricular diastolic filling evaluated by tissue Doppler echocardiography (E' / A' < 1 ) had higher values of NT-proBNP (NS). Other echocardiographic parametres did not show any dependence on NT-proNBP values, including the morphology or atrial defects in the right heart sections which are most conspicuous in an echocardiographic examination. CONCLUSION: Asymptomatic patients after radical correction of TOF have higher values of NT-proBNP (167.95 +/- 91.75 pg/ml). At the same time, the increase closely correlates with the detection of a global right ventricle systolic dysfunction evaluated by S (TDI). On the other hand, there is often no correlation between highly conspicuous changes in the morphology of the right heart compartments or residual or postincision defects of the pulmonary valve on the one hand and increased NT proBNP on the other. The S measurement has the potential to become routine examination in patients after radical correction of TOF for timely detection of right ventricular systolic dysfunction. Precise prognostic and primarily therapeutic impact of the pathologic finding still needs to be determined. PMID- 17419172 TI - [The impact of secondary insertion of ePTFE-coated stent on sustainable TIPS patency]. AB - OBJECTIVE: Retrospective evaluation of the effect of secondary insertion of ePTFE coated stent in the treatment of TIPS dysfunction versus other current options (simple angioplasty, insertion of additional non-coated stent). PATIENT SET AND METHODOLOGY: From the beginning of 2000 to the end of 2004, there were 121 interventions for TIPS dysfunction performed in our centre in which a non-coated stent was used to make up the shunt at the time of intervention. Depending on the type of intervention, the patient set was divided in 4 groups: simple angioplasty (52 cases, 43%), insertion of non-coated stent (35 cases, 28.9%), insertion of non-dedicated ePTFE-coated stent (15 cases, 12.4%), and insertion of dedicated ePTFE-coated stent (19 cases, 15.7%). All patients were monitored on a regular basis after the intervention for shunt patency with the use of clinical examination and Doppler ultrasonography, or also portal venography. Primary shunt patency after the intervention was evaluated in all four groups by Kaplan-Meier analysis. The primary shunt patency results after the intervention were compared with the use Cox F text and logrank test. RESULTS: The intervention was successful in 120 cases (the overall technical success rate of all interventions was 99.2%). The primary shunt patency was 49.7 % after 12 months and 25.3 % after 24 months following sole angioplasty intervention; 74.9% after 12 and 64.9% after 24 months following intervention involving the insertion of non-coated stent; 75.2 % after 12 months and 64.5% after 24 months following intervention involving the insertion of non-dedicated ePTFE-coated stent, and 88.1% after 12 months and 80.8% after 24 months following intervention involving the insertion of a dedicated ePTFE-coated stent. A statistically significant improvement in shunt patency was obtained in the group of interventions involving the insertion of dedicated ePTFE-coated stent and in the group of interventions involving the insertion of non-coated stent as compared with the group of interventions involving sole angioplasty (p < 0.01). CONCLUSION: From among all the currently used methods of therapeutic intervention for TIPS dysfunction, the best, the best subsequent TIPS patency was obtained after intervention involving insertion of dedicated ePTFE-coated stent. PMID- 17419173 TI - [Low-dose thalidomide in refractory and relapsing multiple myeloma]. AB - INTRODUCTION: Thalidomide is one of the drugs which are newly used in the therapy of multiple myeloma. Its immunomodulating action and a number of additional effects have been proven in the treatment of advanced and refractory stage of the disease. However, the best dosing scheme has not yet been discovered and is the subject of research in a number of clinical studies today. PATIENTS AND METHODS: On a retrospective basis, we evaluated results for 59 patients with multiple myeloma who were treated with thalidomide in our facility (median dose of 100 mg), in monotherapy or in combination with corticosteroids, between 2000 and 2005. The objective was to determine the percentage of responses to treatment in patients at different stages of the disease. Response to treatment was evaluated in accordance with EBMT standards. RESULTS: Thalidomide was used as 2nd line therapy (1st relapse or primarily resistant disease) in 59% of cases (35 patients), and as 3rd line therapy (2nd relapse) in 37 % of cases (22 patients). 2 patients were receiving thalidomide as 4th line therapy. None of the patients had taken thalidomide as part of previous treatment. The response rate at 1st relapse (CR - complete remission, PR - partial remission, MR - minimum response) was 60% (21 patients), of which CR was recorded in 2 patients (6%), PR was recorded in 12 patients (35%) and MR in 6 patients (17%). The response rate at 2nd relapse was 45% (10 patients), of which CR was recorded in 3 patients (14%), PR in 1 patient (5%) and MR in 5 patients (23%). Even though we did not record any statistically significant difference in the response of the evaluated group of patients to the treatment with thalidomide at 1st and 2nd relapse, a higher percentage or progression during treatment (32% vs. 14%) was observed in patients at 2nd relapse. 2 patients treated with thalidomide at 3rd relapse did not have a satisfactory response to the treatment (progression or short stabilisation of the disease with subsequent progression). Only 3 patients (5%) of the evaluated group had to discontinue the treatment due to severe adverse events (neuropathy, allergic reaction, leukopenia). The follow up time for Thalidomide therapy ranged between 3 and 62 months for both groups (with a median of 10 months) and spanned from 3 to 60 months at 1st relapse (median of 12 months) and from 3 to 57 months at 2nd relapse (median of 6 months). No statistically significant differences were observed between the 1st and 2nd relapse groups of patients in terms of response rates or length of effect. CONCLUSION: Thalidomide is highly efficient in the treatment of multiple myeloma. The results of study document effectiveness of thalidomide regardless of the disease stage. Comparison of study data with the results of other studies shows that the effectiveness of lower doses we used is comparable with that of higher doses. The fact that lower doses of thalidomide reduce the incidence of adverse events is a clear advantage. PMID- 17419174 TI - [Pancreaticopleural fistula]. AB - Pancreatic diseases are often accompanied by pleuropulmonal complications. Acute pancreatitis may induce a number of various pathological findings in respiratory tract including hypoxemia, decrease of transfer-factor (DLCO), decrease of transfer-coefficient (DLCO/VO), decrease in forced expiratory flow 25%- 75% of forced vital capacity (FEF25-75%), elevated and/or immobile diaphragm, basal atelectasis, unilateral or bilateral pulmonary infiltrations, mediastinal pseudocyst and pleural effusion. Acute respiratory distress syndrome (ARDS) is the most dangerous complication of acute pancreatitis. Large, recurrent pleural effusion is sometimes present in chronic pancreatitis, typically with a very high concentration of amylase in pleural fluid. Pancreaticopleural fistula (PPF) is the most common cause of this type of pleural effusion. We describe a study group of 3 patients with PPF and pleural effusion, their clinical symptoms, diagnostics and management. PMID- 17419175 TI - [Paradoxical embolization and patent foramen ovale in scuba divers: screening possibilities]. AB - INTRODUCTION: The cause of decompression sickness (DCS) in scuba-divers is bubble formation in tissues and in venous blood during ascent. Divers with patent foramen ovale (PFO) have an increased risk of paradoxical embolization to the brain or other vital organs. The aim of our study was to assess the incidence of PFO in scuba-divers with DCS, to compare the group with asymptomatic controls, and to evaluate ultrasound contrast methods suitable for screening. METHODOLOGY: We examined 28 scuba-divers (more than 100 dives). The right-to-left shunt detection was performed by bubble contrast transthoracic echocardiographic examination (TTE) and transcranial Doppler sonography over arteria cerebri media (TCD) in all divers. In divers with shunting, transoesophageal echocardiography (TEE) was performed to prove PFO. RESULTS: 15 divers had DCS associated with the ascent. In this group, PFO was diagnosed in 53% (8/15). The symptoms of all of them retrospectively were of paradoxical embolization (neurological form of DCS). In the group of asymptomatic divers, PFO was proven on the basis of right-to-left shunt screening in 1 diver (8% 1/13). TCD proved right-to-left shunt in all divers with PFO. CONCLUSION: DCS can unmask a so far asymptomatic intracardiac right-to-left shunting. PFO is a risk factor for paradoxical embolization in divers. TCD is suitable for screening; TEE is a gold standard in PFO detection. Our results showed that PFO detection is a useful clinical tool after repeated DCS and in all frequent divers and instructors. PMID- 17419176 TI - [Clinically manifest gastrointestinal bleeding in patients subject to coronary angiography]. AB - PATIENT SET AND METHODOLOGY: The authors evaluated the incidence of acute bleeding from the upper gastrointestinal tract in 5,955 patients (of which 3,684 men and 2,271 women) during hospitalisation for coronary angiography, and the incidence of potential sources of bleeding from the upper gastrointestinal tract in the patients without bleeding. RESULTS: Bleeding occurred in 9 persons, within 3.4 +/- 3.6 days of the coronary angiography (the median of 1.0 day), with a 33% mortality rate. An ulcer of the duodenal bulbus or bulbitis were detected in four cases (44%), esophagitis in one case (11%), esophageal varices in one case, stomach carcinoma in one case, and the source of bleeding could not be detected in 2 cases. Patients with bleeding were significantly older than those without bleeding (73.6 +/- 4.4 years vs. 65.8 +/- 10.6 years, p < 0.001). Treatment with clopidogrel or abciximab was not associated with a higher incidence of bleeding (p > 0.05). In 42 patients without bleeding, the following pathologies were detected by gastroscopy: esophagitis (31%), mouth and stomach ulcers (36%), duodenal ulcers (21%), (12%), esophageal varices (2%) The incidence of mouth ulcers, stomach ulcers and duodenal ulcers was significantly higher in patients taking acetylsalicylic acid on a regular basis (p < 0.025). CONCLUSION: Bleeding from the upper gastrointestinal tract is not frequent shortly after coronary angiography, but the related mortality is high. The most frequent source of bleeding are duodenal peptic lesions most likely caused by previous treatment by acetylsalicylic acid. PMID- 17419177 TI - [Long term effectiveness of surgical cryoablation for chronic atrial fibrillation in patients undergoing surgery for severe mitral valve regurgitation]. AB - OBJECTIVE: The objective of the study was to evaluate effectiveness of surgical cryoMAZE ablation for chronic atrial fibrillation (AF) in patients undergoing mitral valve surgical intervention. METHODOLOGY: Forty-seven patients (31 females), aged 67.3 +/- 7.3 years who underwent surgical intervention for severe mitral regurgitation were studied. Mitral valvuloplasty was performed in 21 patients, and mitral valve replacement in 26 patients. Combined procedure was employed in 35 patients; simultaneous aortocoronary bypass was performed in 16 patients, tricuspid valvuloplasty (TVP) in 5 patients, and aortic valve replacement (AVR) in 5 patients. RESULTS: The mean follow-up time was 19 +/- 10 months. After 6 or 12 months 36 or 32 patients were seen and 23 (64%) or 22 (69%) of them were in stable sinus rhythm (SR), respectively. In the subset of 24 patients with simultaneous intervention on a different valve (TVP or AVR), after 6 or 12 months, 14 (74 %) or 15 (83 %) patients had stable SR, respectively. In the follow-up period, 2 patients underwent successful catheter ablation for type I atrial flutter or for a residual left atrial atypical flutter. CONCLUSION: In the study using the method of cryoMAZE ablation for chronic AF performed during the mitral valve surgical intervention, a long-term stable SR was achieved in a high proportion of patients, particularly in patients with simultaneous intervention on two or three different valves. PMID- 17419178 TI - [Dysfunction of transjugular intrahepatic portosystemic shunt (TIPS) and applicable solutions]. AB - A synoptic article providing a comprehensive view of TIPS dysfunction. The article covers current terminology, definition, etiology, pathogenesis, diagnostics, therapy and prevention of TIPS dysfunction. PMID- 17419179 TI - [Encapsulating peritoneal sclerosis]. AB - Encapsulating peritoneal sclerosis (EPS) is a rare complication of peritoneal dialysis (PD) characterized by extensive peritoneal fibrosis and bowel obstruction. Opinions on its etiopathogenesis diverge. It appears that the performance of peritoneal dialysis itself and the exposition of the peritoneum to non-physiological influences (the incidence increases in proportion to the length of peritoneal dialysis treatment) is an important factor. Yet there is also patients' individual sensitiveness, which apparently depends on individual immunological response. EPS causes severe changes in peritoneal anatomy and functions, resulting both in the loss of the transport function of the peritoneum, and in bowel malfunction and serious malnutrition. Imaging methods (ultrasound, computer tomography) are very important in diagnosing EPS; the diagnosis is confirmed histologically. No laboratory markers have been discovered yet to allow for timely indication of EPS development. There are different treatment strategies, including different surgical interventions (liberation of bowel loops, peritoneum resection) and the effort to influence the immune process. Corticosteroids remain the medication of choice; also promising is Tamoxifen. In spite of that, EPS mortality is still very high, ranging between 56 and 93%. The disease often progresses even after peritoneal dialysis has finished. The article summarises the existing knowledge about encapsulating peritoneal sclerosis, its diagnosis and therapy. PMID- 17419180 TI - [Fever of unknown origin--etiology and diagnostic algorithm]. AB - Fever of unknown origin is a frequent and significant diagnostic problem often faced by physicians. The first part of the text is dedicated to its definition and wide-ranging aetiology. On the one hand, fever may be a banal and benign condition, on the other, it can be the symptom of a life threatening disease. The second part presents our suggestions for diagnostic approach to fever of unknown origin. We believe this text may become a useful tool for this extremely complex and interesting differential diagnostic. In view of extension and complexity of the topic, the text of this part is presented in full. PMID- 17419181 TI - [Czech Atherosclerosis Society Guidelines for the diagnosis and treatment of dyslipidemias in adults]. AB - The present guidelines are based on the recommendations published in 2005 entitled "Prevention of Cardiovascular Diseases in Adulthood" summarizing the conclusions of nine Czech medical societies and agree with them in the assessment of individual risks of mortality from cardiovascular disease (CVD) according to SCORE tables. They reflect new research data in pathophysiology of dyslipidemias (DLP) and particularly the results of recent clinical trials of lipid-lowering therapy and their meta-analyses. They establish priorities for the screening and management of DLP, present suitable diagnostic methods, additional investigations of potential use in risk assessment, including some emerging risk factors and detection of sub-clinical atherosclerosis in persons in a moderate-risk category. Major changes include a lower LDL-cholesterol treatment target (< 2.0 mmol/L for all CVD individuals) and a possible use of apolipoprotein B as a secondary target in selected persons (< 0.9 g/L in high risk without CVD, < 0.8 g/L for CVD patients) and nonHDL-cholesterol (< 3.3 mmol/L in high risk without CVD, < 2.8 mmol/L for CVD patients). Therapy of individual DLP phenotypes (monotherapy and combination therapy) as well as basic principles for control examination at lipid lowering medication are described. Recommended therapeutic lifestyle changes are shown. Enclosed are five annexes: DLP diagnosis; causes of secondary DLP; additional investigations of potential use in risk stratification; familial hypercholesterolemia; list of recommended foods; two variants of SCORE tables for risk assessment for the Czech Republic; the scheme of recommended procedures and treatment algorithm in DLP asymptomatic individuals. PMID- 17419182 TI - [Dialogue of a haematologist with an internist. The issue of specialisation and information in medicine]. PMID- 17419183 TI - (1 --> 3)-beta-D-glucan--a new marker for the early serodiagnosis of deep-seated fungal infections in humans. AB - Until recently, the diagnosis of systemic mycoses was mainly based on traditional methods producing late and inconclusive results. Currently used methods of serological diagnostics are generally based on detection of cell wall components of selected pathogenic fungal species--mannoproteins, functioning as a antigenic markers. There are big hopes for adaptation of commercially available assays to detect (1 --> 3)-beta-D-glucan because of the fact that its presence in blood and normally sterile body fluids should lead to initiation of the diagnostic workup of invasive fungal infection. Monitoring (1 --> 3)-beta-D-glucan antigenemia is useful in predicting the therapeutic outcome of patients with invasive aspergillosis and in combination with galactomannan detection to identify false positive reactions. The simultaneous use of both tests is probably more pertinent for the differentiation between yeast and mould infections. PMID- 17419184 TI - Genetic and physiological regulation of bacterial endospore development. AB - Bacterial endospores are complex structures residing inside endospore-forming, mainly gram-positive bacteria. The process of sporulation is considered a simple example of cell differentiation. Endospores enable the organism to resist environmental stresses. Sporulation can be divided into several stages, from axial DNA filamentation to mother cell lysis. The structure and formation of an endospore is an attractive model for the assembly of complex macromolecular structures during development. The expression of genes involved in sporulation is compartmentalized and different sets of genes are expressed in the prespore and mother cell, this being associated with the subsequent activation of four sporulation-specific sigma factors. Their synthesis and activity are tightly regulated and the regulatory mechanisms have overlapping roles. PMID- 17419185 TI - Molecular characterization of class 1 integrons in clinical strains of Salmonella typhimurium isolated in Slovakia. AB - The presence of class 1 integrons was investigated in 156 epidemiologically unrelated Salmonella Typhimurium isolates. Of these 156 isolates, 70 were of definitive phage type DT104 and 86 were strains of various phage type, RDNC and untypable, designated here as non-DT104 strains. Integrons were found in 47 of DT104 isolates (67.1%), while in all strains with characteristic pentaresistance (R-type ACSSuT) two integrons 1.0 kb and 1.2 kb in size were found. Among 86 non DT104 strains, integrons with sizes of 1.6 kb and 1.9 kb in four multidrug resistant strains DT193 and U302 were found. The integrons from selected strains were further sequenced and the aadA1, aadA2, dhfr1, dhfr12 and bla(PSE) genes were found embedded in cassettes. PMID- 17419186 TI - Toxoplasma gondii: inhibition of the intracellular growth by human lactoferrin. AB - In the present paper we focused on the potential role of human lactoferrin for the intracellular replication rate of T. gondii BK tachyzoites following our earlier observation that Toxoplasma gondii was able to bind human lactoferrin but not serum transferrin. The study was performed in vitro on human CaCo-2 epithelial cells and mouse L929 fibroblasts. We found that the multiplication of the parasite was inhibited by lactoferrin in both cell lines used. However, the direct cytotoxic effect on the parasite and the host cells was not observed. The intracellular growth of T. gondii was not affected when tachyzoites or host cells were only pre-coated with human lactoferrin. The results suggest that lactoferrin does not influence parasites penetration into host cells but could trigger unknown antiparasitic mechanisms in the infected cells. PMID- 17419187 TI - Presence of Bartonella spp. in various human populations. AB - Bartonella spp. bacteria are significant human pathogens and the agents of bacterial zoonosis acquired from an animal companion. The aim of the study was to determine the seroprevalence of two of the most common Bartonella species B. henselae and B. quintana in various human populations. The studied groups included: alcoholics, intravenous drug users, veterinarians, cats' owners. Blood samples were collected and cultured on chocolate agar plates and in mouse fibroblasts L-929 cell line culture. The levels of Bartonella henselae IgM and IgG antibodies were determined by indirect immunofluorescence assay. Specific B. henselae IgG were detected in 48.3% of homeless alcoholics, in 45.0% veterinarians and in 53.3% cats' owners. The differences in the prevalence of B. henselae antibodies between the studied groups and a control group were statistically supported. No homeless intravenous drug users had specific B. henselae and B. quintana antibodies. High titers of B. quintana IgG antibodies were detected in two homeless alcoholics. Bartonella spp. was cultured on chocolate blood agar plates from blood samples from 2 alcoholics. The isolates were identified as B. henselae by PCR amplification of the riboflavin synthase gene (ribC). The results prove that B. henselae and B. quintana, emerging human pathogens, are present and widely distributed in Poland in such specific risk groups as: alcoholics, veterinarians and cats' owners. PMID- 17419188 TI - Killer activity of yeasts isolated from natural environments against some medically important Candida species. AB - Twenty-five yeast cultures, mainly of human origin, belonging to four pathogenic yeast species--Candida albicans, Candida krusei, Candida parapsilosis, and Candida tropicalis were tested for their sensitivity to ten basidiomycetous and eleven ascomycetous yeast species isolated from the water and soil environments and from tree leaves. The best killer activity among basidiomycetous species was exhibited by Rhodotorula glutinis, and R. mucilaginosa. The other carotenoid producing species Cystofilobasidium capitatum, Sporobolomyces salmonicolor, and S. roseus were active only against about 40% of the tested strains and exhibited weak activity. The broadest killer activity among ascomycetous yeasts was shown by the strains Pichia anomala and Metschnikowia pulcherrima. The species Debaryomyces castellii, Debaryomyces hansenii, Hanseniaspora guilliermondii, Pichia membranifaciens, and Williopsis californica did not show any killer activity. The best killer activity exhibited the strains isolated from leafy material. The lowest activity pattern was found among strains originating from soil environment. PMID- 17419189 TI - Molecular analysis of bacterial community diversity in sequencing batch reactor (SBR) operating in autotrophic conditions. AB - In the present study the bacterial community structure in a sequencing batch reactor (SBR) operating in autotrophic conditions was determined in relation to temporal variations of NH4-N, NO2-N, NO3-N levels in the effluent. Bacterial richness and composition were determined by PCR-DGGE (Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis) of 16S rRNA and amoA genes, and RISA (Ribosomal Intergenic Spacer Analysis) technique. The applied approaches revealed that the composition of bacterial population in the studied SBR varied in time. A positive correlation between Shannon index of ammonia-oxidizing bacteria (AOB) diversity and ammonia level in the effluent was observed. The variation of total bacterial diversity did not reflect the changes in nitrification efficiency. PMID- 17419190 TI - Geomicrobiological aspects of the oxidation of reduced sulfur compounds by photosynthesizing bacteria. AB - The activity of photosynthesizing sulfur bacteria in a continuous culture was studied. The bacteria were isolated from the natural environment with the use of the Winogradski column. Isolated bacteria were cultured in synthetic medium and in the effluent from the column containig HS-. Sulphides, the main product of reduction of sulfates in phosphogypsum, were used by green sulfur bacteria in the photosynthetic column. Almost 70% reduction of the concentration of sulfides was observed. After the experiment, diffractometric methods where employed to analyze the sediment from the column. PMID- 17419191 TI - Avoid quick fixes for improvement: don't let surveyors find reoccurring problems. PMID- 17419192 TI - Coming soon: better data on hospital-acquired conditions. PMID- 17419193 TI - State goes extra mile with new charity policy. PMID- 17419194 TI - WA hospitals adjust to meet aid guidelines. PMID- 17419195 TI - Enlist staff to find system flaws before errors occur. PMID- 17419196 TI - Uncover opportunities using comparative data. PMID- 17419197 TI - [Rotational landmarks and total knee arthroplasty in osteoarthritic knees]. AB - OBJECTIVE: To investigate the rotational mismatch of total knee replacement with medial 1/3 of tibial tuberosity as bony landmark in osteoarthritic patients with varus or valgus deformity. METHODS: Axial images on computed tomography of 62 knees (including 55 varus deformities and 7 valgus deformities) in 32 Chinese osteoarthritic patients who had total knee arthroplasty were analyzed, compared with that of 10 healthy knees. On images of the distal femur, the angle between the lines of surgical epicondylar axis (SEA) and posterior condylar axis was measured as posterior condylar angle (PCA), and on images of the proximal tibia, a baseline for the anteriorposterior axis of each component was drawn based on the SEA for the femur and the medial 1/3 of the tibial tuberosity for the tibia. The angle between these lines (Angle a) was defined as the rotational mismatch between the components when they were aligned to the anatomic landmarks of each bone. RESULTS: The sulcus of medial epicondyle of femur could be identified on CT images of over 80% osteoarthritic knees; the median value of PCA was +2.36 degrees, with an individual variation of 0 degree to +7.5 degrees. Angle a was +6.45 +/- 3.68 degrees (range, 0 degree to +11.8 degrees) in 10 healthy knees, which increased significantly to +10.85 +/- 10.47 degrees (range, 0 degree to +28.1 degrees) in 55 varus knees (P < 0.05), which also increased significantly to +11.6 +/- 7.3 degrees (range, -6.5 degrees to +26.8 degrees) in 7 valgus knees (P < 0.05). CONCLUSION: With the medial 1/3 of the tibial tuberosity as the rotational landmark for the tibial component, there was a tendency to align the tibial component in external rotational position relative to the femoral component in knees with normal alignment, the rotational mismatch increased in Chinese osteoarthritic knees with varus and valgus deformity. PMID- 17419198 TI - [Short-term effect of rotating hinge knee prosthesis]. AB - OBJECTIVE: To discuss short-term effect of rotating hinge knee prosthesis. METHODS: From July 2002 to April 2005, 17 cases of severe knee joint deformity and instability received rotating hinge knee prosthesis for total knee arthroplasty. There were 8 males and 9 females, aging from 4l to 79 years. The left joints were involved in 10 cases and right joint in 7 cases. All patients were admitted because of pain. The course of disease was from 1 to 7 years. There were 10 cases of osteoarthritis, 5 cases of rheumatoid arthritis, 1 case of traumatic arthritis after operation of left femur fracture,and 1 case of traumatic arthritis with injury of anterior cruciate ligament, meniscus medialis and medial collateral ligament after operation of left fracture of tibial plateau. According to HSS (hospital for special surgery) scoring system, the preoperative score was 36 to 58 with an average of 48.6. The preoperative flexed motion range of articulation was 21 degrees to 80 degrees with an average of 57.4 degrees. RESULTS: All patients were followed up from 7 months to 3 years with an average of 23. 6 months. There were no complications of thrombogenesis of veins of lower extremity, pulmonary embolism, palsy of peroneal nerve, fracture, and breakage of extended knee structure. Infection occurred in 1 case at 3 months postoperatively, the prosthesis was dislodged, antibiotic-impregnated cement was filled with knee joint, two-stage arthroplasty was expected. At the last follow up, the HSS score was 78 to 98 with an average of 91.1 in 16 patients. The flexed motion range of articulation was 75 degrees to 100 degrees with an average of 85.2 degrees at 2 weeks postoperatively. The flexed motion range of articular was 85 degrees to 123 degrees with an average of 108.3 degrees at the last follow-up. There were significant differences in HSS score and motion range of articular between preoperation and the last follow-up (P < 0.05). CONCLUSION: The short term outcome of rotating hinge knee prosthesis is good and a long term follow-up is necessary. PMID- 17419199 TI - [Analysis of factors relating to clinical outcomes after total knee replacement]. AB - OBJECTIVE: To analyze the outcomes and complications after total knee replacement (TKR) with posterior stabilized prosthesis (PS) and to investigate the influencing factors relating to outcomes. METHODS: From January 1998 to August 2004, 60 cases (74 knees) of osteoarthritis underwent TKR with PS. The outcomes were evaluated according to the HSS (hospital for special surgery) scoring. The difference in outcomes between patients with post-operative complications and without complications were compared. Pearson correlation was used to analyze postoperative outcomes and the pre-operative factors relating to patients. RESULTS: All 74 knees were followed up 42.5 months (24 to 94 months). The scores for HSS, pain, function, ROM muscle strength, flexion deformity and stability of knees after operation were 84.2 +/- 14.2, 25.7 +/- 6.9, 17.9 +/- 4.3, 13.1 +/- 2.0, 9.2 +/- 0.8, 8.1 +/- 0.4 and 9.3 +/- 0.1 respectively. They were improved to some extents, especially pain alleviation was remarkable. The excellent and good rate for outcome assessment was 90.5%. Among 74 knees, 10 cases suffered from post-operative complications, including 1 case of common peroneal nerve paralysis, two cases of wound faulty union, one case of wound infection, one case of joint infection, one case of stiff knee, two cases of deep vein thrombosis and 2 cases of patellofemoral joint complications. The excellent and good rate of outcome in patients with complications (60%) was much lower than that in patients without complication (95.3%),and there was significant difference between them (P < 0.05). Analysis for correlation showed that postoperative HSS score was positively correlative with the postoperative HSS score, pain and function score of knees. The correlation value was 0.523, 0.431 and 0.418 respectively (P < 0.01). Whereas, postoperative HSS score was not correlative with ROM, muscle strength, flexion deformity, stability of knee, age, weight and body mass index (P > 0.05). CONCLUSION: TKR with PS is an effective method for severe osteoarthritis. The outcomes after TKR have a positive correlation with the HSS score, pain and function score of knees before surgery. Complications associating with surgery have a negative influence on outcomes. PMID- 17419200 TI - [Replacement of invert-type artificial total scapula and shoulder joint prosthesis for malignant tumor]. AB - OBJECTIVE: To estimate the clinical curative effect of replacement of invert-type artificial total scapula and shoulder joint prosthesis and reserving arm with rehabilitation of function in the treatment of.malignant tumor in shoulder. METHODS: From February 2001 and November 2004, five youth patients with primary malignant shoulder tumors were treated operatively by resection of neoplasms thoroughly, replacement of invert-type artificial total shoulder blade and joint prosthesis, the functional reconstruction. Of them, there were 4 males and 1 female, aging from 19 to 26 years with an average of 23.6 years. Two cases were diagnosed as having osteosarcoma, one as having chondrosarcoma, and 2 as having Ewing sarcoma. After operation, the upper limbs was immobilized for 3 weeks. The rehabilitation training including passive exercise and initiative exercise. RESULTS: The average operative time was 425 min (380 to 530 min), and the blood loss ranged from 1250 ml to 1900 ml (1540 ml on average). The follow-up ranged from 7 to 52 months, with an average of 24.6 months. Postoperative complication included 1 case of pneumothorax, one case of shoulder incision skin part necrosis and 1 case of clavicle stump raising and pierce skin with shallow infection. No complication of postoperative incision deeply infection, nerve damage and prosthesis exposure or dislocation occurred. According to the scoring system of JOA (Japan orthopaedics association), the average score was 65 (60 to 72). The flexion and extension function of elbow joint recovered to normal. CONCLUSION: The replacement of invert-type artificial total scapula and shoulder joint prosthesis is an efficacious method for the treatment of malignant tumor in shoulder. There are advantages of numerous adaption, wide range of motion and good stability. It can not only reserve arm but also rehabilitate function. PMID- 17419201 TI - [Short-term follow-up of ribbed anatomic cementless total hip arthroplasty]. AB - OBJECTIVE: To summarize the clinical outcome of the Ribbed anatomic cementless total hip arthroplasty (THA) in the treatment of hip-joint disease. METHODS: From January 2001 to June 2005, 34 patients (38 hips) with hip-joint disease were treated with Ribbed anatomic cementless THA. Their ages ranged from 29 to 55 years with an average age of 42.7 years. The disease course was from 3 to 18 years. Among these cases, there were 7 cases (7 hips) of femoral neck fracture, 5 cases (5 hips) of traumatic arthritis after fracture of acetabulum, 15 cases (16 hips) of necrosis of the femoral head and 7 cases (10 hips) of ankylosing spondylitis. Four patients were operated on both hip joints. The average Harris hip score was 38.6 (25-57) before operation. RESULTS: Twenty-one patients(23 hips) were followed up 8-61 months with an average of 35 months. The Harris hip score was 76-98 after operation with an average of 92.3, showing significant difference when compared with that before operation (P < 0.05). The excellent and good result was achieved in 93.5% of patients. Radiographs showed no prosthetic osteolysis and no evidence of loosening. Pain in the thigh occurred in 4 patients,and it can be relieved by using non-steroid anti-inflammatory drug. CONCLUSION: Ribbed anatomic cementless THA has good clinical and radiographic results in treating patients with hip-joint disease. PMID- 17419202 TI - [Bioabsorbable interference screw for ligament reconstructions in the knee]. AB - OBJECTIVE: To evaluate the clinical results of bioabsorbable interference screw in knee ligament reconstruction. METHODS: From April 2002 to August 2004, 39 patients with knee ligament injury were treated, including 33 males and 6 females with a median age of 25 years (15 to 65 years). The involved ligament included 29 anterior cruciate ligament of knee (ACL), 6 posterior cruciate ligament of knee (PCL), 4 combined ACL and PCL, 3 medial collateral or lateral collateral ligaments of knee point and 1 posterolateral complex injury of knee. All of patients underwent anatomic ligament reconstructions under arthroscopy or open surgery by autografts and fixation with bioabsorbable interference screw. RESULTS: All 34 patients were followed up 6 to 28 months (mean 13.7 months). The patients were evaluated by Lysholm knee functional scales, the knee scores were 43.6 +/- 13.4 before operation and 85.4 +/- 16.3 after operation, showing significant difference (P < 0.05). There was no limitation of range of motion and loosening of the screw. However, one case suffered from infection, and 3 cases suffered from effusion and synovitis after surgery and recovered after management. CONCLUSION: Bioabsorbable interference screw fixation is a reliable method in knee ligament reconstruction and is effective to restore knee joint stability. PMID- 17419203 TI - [Treatment of posterolateral corner injury of knee joint with anatomical reconstruction]. AB - OBJECTIVE: To discuss the diagnosis and treatment of posterolateral corner injury of the knee joint, and to evaluate the clinical results after anatomical reconstruction. METHODS: Sixteen cases of posterolateral corner injury of the knee were treated, being accompanied with grade II or above existing serious varus or external rotation. At a mean follow-up of 13 months (7 to 18 months), we evaluate the clinical results using Lysholm and IKDC (international knee documentation committee) scoring systems. Among them, there were 14 males and 2 females aging 19-46 years (mean 38 years). It was 4-12 d from injury to operation. On conditions of anesthesia, varus stress test (VST) showed that instability of varus was more than 10 degrees under the 30 degrees and 0 degrees of genuflexion respectively and external rotation test showed that instability of external rotation was more than 10 degrees when compared with normal side. Five cases complicated by rupture of anterior cruciate ligament, 8 cases by rupture of posterior cruciate ligament and 3 cases by rupture of both ligaments. RESULTS: All patients were followed 7 to 18 months and no patients had instability of the knee. The VST showed that no abnormal lateral instability was found at 0 degrees of flexion except 3 cases at 30 degrees with less than 5 degrees. Cooper test showed that no rotational instability at 90 degrees except 1 cases at 30 degrees with less than 5 degrees. The range of motion of the knee was from 75 degrees to 130 degrees of flexion(mean 118 degrees)and from 0 degrees to 5 degrees of extension. Posteroperative scores were from 54 to 96 according to Lysholm scoring systems and from 46 to 94 according to IKDC scoring systems. CONCLUSION: Although the overall incidence of posterolateral corner injury is less than that of other ligament injuries of the knee, we should pay much attention to posterolateral corner injury because of its important role in resisting posterior tibia translation, external rotation and varus of knee. This injury can result in a significant degree of disability for patients. PMID- 17419204 TI - [Effect of calcaneocuboid arthrodesis on three-dimensional kinematics of talonavicular joint]. AB - OBJECTIVE: To discuss the effect of the calcaneocuboid arthrodesis on three dimensional kinematics of talonavicular joint and its clinical significance. METHODS: Ten fresh-frozen foot specimens, three-dimensional kinematics of talonavicular joint were determined in the case of neutral position, dorsiflexion. plantoflexion, adduction, abduction, inversion and eversion motion by means of three-dimensional coordinate instrument (Immersion MicroScribe G2X) before and after calcaneocuboid arthrodesis under non-weight with moment of couple, bending moment, equilibrium dynamic loading. Calcaneocuboid arthrodesis was performed on these feet in neutral position and the lateral column of normal length. RESULTS: A significant decrease in the three-dimensional kinematics of talonavicular joint was observed (P < 0.01) in cadaver model following calcaneocuboid arthrodesis. Talonavicular joint motion was diminished by 31.21% +/- 6.08% in sagittal plane; by 51.46% +/- 7.91% in coronal plane; by 36.98% +/- 4.12% in transverse plane; and averagely by 41.25% +/- 6.02%. CONCLUSION: Calcancocuboid arthrodesis could limite motion of the talonavicular joints, and the disadvantage of calcaneocuboid arthrodesis shouldn't be neglected. PMID- 17419205 TI - [Study on biomechanical model of rabbit femur]. AB - OBJECTIVE: To discuss the method of constructing biomechanical model of rabbit femur. METHODS: The sample of rabbit femur was prepared as follows: firstly femur section images were obtained, then the image was put into the computer and processed to get the boundary contour line; secondly, through programming the contour line coordinate for modeling was obtained, then the data were put into the model software to find the three-dimensional entity model. RESULTS: Whole three-dimensional model of rabbit femur was constructed. It simulated actually dissection form of femur. CONCLUSION: The establishment of the model lays a foundation for ascertaining optimal parameter of vibration improving bone minery density by finite element analysis. PMID- 17419206 TI - [An experimental study on osteocyte apoptosis in steroid-induced early osteonecrosis of femoral head]. AB - OBJECTIVE: To explore the significance of osteocyte apoptosis in steroid-induced osteonecrosis of the femoral head. METHODS: Sixty New Zealand rabbits were divided into experimental group and control group (n=30). The experimental group was given 10 ml/kg of horse serum intravenously 2 times at 2 weeks intervals and an intraperitoneal injection of 45 ml/kg x d of methylprednisolone acetate for 3 days; the control group was given equal isotonic Na chloride. Osteocyte apoptosis was observe by means of TUNEL. RESULTS: The number of apoptosis in the experimental group (112.33% per hundred +/- 26.12% per hundred) was significantly higher than that in the control (47.01% per hundred +/- 22.95% per hundred) (P < 0.01) in the 4th week. With time, osteocytes apoptosis progressively increased. In the 6th and 8th weeks, the percentage of empty osteocyte lacunae in the experimental group (17.23% +/- 3.44%, 28.56% +/- 3.45%) was significantly higher than that in the control group (11.29% +/- 2.89%, 11.26% +/- 2.75%, P < 0.05). The transmission electron microscope showed that the characteristics of osteocyte apoptosis included intact nuclear membrane,c omdensed chromatin and increased electron dense. CONCLUSION: Osteocytes apoptosis may play a key role in the process of steroid-induced early osteonecrosis of the femoral head. PMID- 17419207 TI - [Treatment of old united lateral condyle fractures of humerus in children]. AB - OBJECTIVE: To analyze the treatment of old united lateral condyle fractures of humerus in children. METHODS: From January 1997 to February 2002, 13 cases of old united lateral condyle fractures of humerus were treated. There were 11 boys and 2 girls. Their age was from 4 to 11 years (7.2 years on average). The primary diagnosis results were 4 cases of degree I fracture, 5 cases of degree II fracture and 4 indefinitely diagnosed cases. Elbow extension disorder occurred in 13 cases, which was limited 30-70. The elbow flexion was limited more than 90 degrees. X-ray films showed 10 nonunion and 3 malunion of fracture. The time from injury to operation was 32 to 81 days (56 days on average) in 12 cases and 6 months in 1 case. All cases were treated by open reduction and internal fixation. One case was treated combined with implanting bone. RESULTS: After a follow-up of 3 to 8 years (5.4 years on average), no cubitus varus or valgus occurred. Five cases had growth disturbances of the lateral condyle of the distal humerus. The movement of these elbow joints improved from 40 degrees to 70 degrees (56 degrees on average). The time of clinical bone union was 6 to 8 weeks and no nonunion and delayed union were observed in X-ray films. CONCLUSION: It is important to accurately diagnose for treatment of lateral condyle fractures of the humerus in children so as to reduce the occurrence of malunion of fracture. It can save the joint function and reduce the cosmetic deformity effectively to operate as early as possible. PMID- 17419208 TI - [Treatment of distal radius fracture by combination of external fixator and volar approach fixation]. AB - OBJECTIVE: To investigate the operative indication and clinical efficacy of combination of external fixator and volar approach fixation in treating distal radius fracture. METHODS: From March 2000 to March 2005, 28 patients with distal radius fracture were treated with combination of external fixator and volar approach fixation. Dorsal external fixator was used to maintain wrist in functional position, combinated volar plate or Kirschner wire fixation after reduction was achieved. Bone graft was necessary if there were severe comminuted cortical bone or compress of cancellous bone. Of 28 patients, there were 21 males and 7 females, aging from 18-54 years with a median age of 41 years. The locations were the left side in 4 cases and the right side in 24 cases. According to typing criterion for AO, 18 cases were classified as C2 and 10 cases as C3. One case was accompanied with dislocation of lunate bone. RESULTS: Among 28 patients, 24 were followed up for an average of 8.5 months. The anatomical relationship of their wrist joint were re-established and retained. Overall good to excellent results were achieved in 87%, excellent in 16 cases, good in 5 cases and fair in 3 cases. CONCLUSION: A combination of external fixator and volar approach fixation is an effective method of treating fractures of the distal radius, because distal radius fracture is unstable or difficult to close reduction. Volar fixation can avoid operative complication, and external fixator can obtain satisfactory reduction and function. PMID- 17419209 TI - [Von Mises stresses research of three-fin acetabular components and pelvis bone with segmental bone defect of acetabulum using rosette strain gages method]. AB - OBJECTIVE: To set up the experimemtal model with superior segmental bone defect in acetabulum and implant the three-fin acetabular component, and examine the Von Mises stresses of pelvis bone with simulating single leg position in vitro using rosette strain gages method (RSGD). METHODS: Four kinds of three-fin components were made based on measurement of diameter, depth and roof thickness of acetabular specimens. These kinds of three-fin acetabular components based on spreaded degrees of lateral fins (0, 36.45 and 60 degrees). The superior segmental bone defect of acetabulum in 4 cadaver pelvis specimens was made,then simulated the actual position to place implant into acetabular bone. A blank control group was set up. After being fit up the 7 sets of rosette strain gages,then specimen to test the pelvic Von Mises stresses simulating the static load of single leg on the mechanic machine. Progressive load was graded into 150, 300, 450, 600, 750 and 900 N. We obtained the strains informations about loaded and un-loaded pelvis bone. RESULTS: Based on computed the strains informations,we were informed the Von Mises stresses about pelvis bone. The maximal Von Mises stresses of selected local position was 6.93 MPa and the minimal Von Mises stresses of selected local position was 1.08 MPa. CONCLUSION: Providing the data about the main part of three-fin acetabular component to optimize spreaded degrees of lateral fins. PMID- 17419210 TI - [Treatment of femoral neck fracture with cannulated screw fixation in young adults]. AB - OBJECTIVE: To evaluate the clinical effect of cannulated screw on treatment of femoral neck fracture (FNF). METHODS: Forty-two FNF patients were treated by using cannulated screw from January 2001 to December 2005. There were 22 males and 20 females with an average age of 41 years (19-59 years). Fracture was caused by traffic accident in 21 cases, by falling from height in 14 cases and by bruise in 7 cases. All cases were fresh fracture. According to Garden criterion for typing, 15 cases were classified as type I , 16 cases as type In and 11 cases as type IV. It was 7 hours to 15 days from injury to operation. RESULTS: The patients were followed up for 1-6 years with an average of 2.5 years. The average fracture union time was 6.5 months. Three patients had ischemic necrosis of femoral head, and loosening and breakage of screw and rob was observed in 1 case. According to Brumback criterion for hip joint function, the result was excellent in 18 cases, good in 20 cases and bad in 4 cases, and the excellent and good rate was 90.4%. CONCLUSION: Cannulated screw fixation is a good method to treat FNF in young adults. It can improve the rate of fracture union and reduce the rate of avascular necrosis of femoral head. PMID- 17419211 TI - [Repairing chronic osteomyelitis complicated by long bone defect in tibia with free segmented-fibula transplantation]. AB - OBJECTIVE: To probe the repair method and effect of free segmented-fibula transplantation to treat chronic osteomyelitis complicated by long bone defect in tibia in the first intention. METHODS: From March 1996 to December 2003, 67 cases of chronic osteomyelitis complicated by long bone defect were reconstructed with vascularized fibula graft after the long inflammable bone and soft tissue focus were resected. Their age ranged from 8 to 42 years. The course of disease was 6 months to 8 years. There were 14 cases of hematogenous osteomyelitis and 53 cases of traumatic osteomyelitis. Of them, 18 cases complicated by fracture of fibula; 21 cases by defect of skin (2 cm x 4 cm-4 cm x 10 cm) and bone exposure; 53 cases by pathological fracture and nonunion; and 46 cases by 1-3 fistula of osteomyelitis. The length of bone defect was from 8 cm to 22 cm (mean 12 cm), and the germiculture results of all cases were positive. Forty-six cases were treated with vascularized fibula graft, the other 21 cases with the skin flap. The segmented-fibula was 10-28 cm, skin flap size was 4 cm x 7 cm-6 cm x 12 cm. RESULTS: After a follow-up of 12-45 months, the healing rate of sinus was 93.5% while the 6.5% remainders healed by the second sinus cleaning-up. The graft bone healed after 4-6 months (mean 4.2 months) by X-ray examination. The limb inflammation was controlled after 2 weeks. All 21 skin flaps all survived and the function recovery of affected limb was 79% of normal limbs according to Enneking evaluation system, but 2 patients occurred secondary fracture. The act or process of augmenting of tibia under 18-year-old cases were sooner than those who were more than 18-year-old. CONCLUSION: It is a choice to repair the chronic osteomyelitis complicated by long bone defect with vascularized fibula graft in the first intention. The operation to reconstruct long bone defect is a good method to control inflammation efficiently, shorten period of treatment and reduce the time of operation. PMID- 17419212 TI - [Effect of blocking screws on breakage of interlocking intramedullary nails]. AB - OBJECTIVE: To explore the effect of blocking screws on the breakage of interlocking intramedullary nails. METHODS: From January 2003 to August 2005, 56 patients with fresh and close fracture of long shafts were treated by interlocking intramedullary nails. Among them, there were 32 males and 24 females, including 26 femoral fracture and 30 tibial fracture. Fracture of femoral and tibial shaft was fixed with interlocking intramedullary nails normally in group I (n=32). And long oblique, spiral, proximal or distal fracture of bone shaft was fixed with interlocking intramedullary nails and blocking screws in group II (n=24). RESULTS: All cases were followed up 12 to 21 months (16 months on average). In group I, fracture healing failed and the intramedullary nails broke in 3 cases, breakage site was the middle femoral fracture area in 1 case and the first distal interlocking nail hole in 2 cases of distal 1/3 tibial fracture. Fractures healed 6 to 12 months after operations in the other cases, but more bony callus occurred in fracture area. In group II, fractures healing and good alignment were achieved, and no breakage was found in all 24 cases. CONCLUSION: The use of blocking screws added to interlocking intramedullary nails could improve the stability of fracture areas distinctively, and hence reduce the breakage of intramedullary nails. PMID- 17419213 TI - [Subtalar arthrodesis and calcaneal thalamus reconstruction for malunion of calcaneal fractures]. AB - OBJECTIVE: To evaluate the clinical results of subtalar arthrodesis and calcaneal thalamus reconstruction for malunion of calcaneal fractures and to discuss the indications and its advantages of the management. METHODS: From December 1994 to January 2006, 84 cases (96 feet) of malunion of calcaneal fractures were treated with subtalar arthrodesis and calcaneal thalamus reconstruction. The L-shaped approach lateral to calcaneus was used. The bone autograft was harvested from iliac crest. Fifty-one cases were male and 33 cases were female, aging from 21 to 58 years (mean 38.5 years). One side was in volved in 72 cases and two sides in 12 cases. The injury was caused by falling from height in 57 cases, by traffic accident in 22 cases and other in 5 cases. It was 6-31 months from injury to operation (mean 9.5 months). RESULTS: Among the patients, all cases were followed up 1 to 132 months. The total excellent and good rate was 87.5% including excellent in 31 feet, good in 53 feet and fair in 12 feet,according to Hindfoot scores system (American Orthopaedics Foot and Ankle Society). The Bohler's and Gissane's angles, the height of calcaneal thalamus and width of calcaneus were significant different from those of preoperation (P < 0.01). CONCLUSION: The treatment by bone autograft combined with subtalar arthrodesis and calcaneal thalamus reconstruction is an effective operation for malunion of calcaneal fractures, with advantages of correcting deformity, restoring the function of hindfoot and relieving the pain of walking. PMID- 17419214 TI - [Clinical manifestation and quantitative analysis on roentgenography of unknownetiology osteoarthritic disease in Chongzhou city of Sichuan province]. AB - OBJECTIVE: To investigate the clinical manifestation and radiological features of unknown-etiology ostearthritic disease in Shigeng village of Chongzhou city in Sichuan province so as to lay a basis for further study. METHODS: The data were analyzed retrospectively. The epidemiology investigation, physical examination, movement functional assessment, lab test and X-ray examination were performed on 116 villagers of 3 years and more than. Quantitative analysis on roentgenography was conducted. RESULTS: Sixty-four villagers had unknown ostearthritic disease,the prevalence was 55.2%; 17 patients were limp, the prevalence of limp was 26.6%, onset age mainly focused under 40 years. Main manifestation of osteoarthritic disease was fixed pain of knee joint or hip joint, and bilateral pain was significantly more than single pain, but local fever and swelling were not obvious. Mild or moderate pain account for 73.5% of patients, and pain was aggravated after physical labor or movement and could be relieved after rest. Joint pain had no influence on joint movement function for long time, leading to a limp due to the joint functional disorders at late stage. Physical examination illustrated the flexion contracture of the hip and limitation of internal and external rotation. The Harris scores were 86.5 +/- 17.1(left hip) and 86.3 +/- 17.2. 46 (right hip), and the excellent or good result was achieved in 46 patients, fair and poor results in 18 patients. The average HSS scores were 88.4 +/- 9.3 (left knee) and 88.2 +/- 11.0(right knee). The excellent or good result was achieved in 61 patients, fair and poor results in 3 patients. The X-ray films showed degenerative disorders of hip joint such as narrowing of the joint space (47.6%), articular surface sclerosis and deformation (30.2%), shallow and increased density acetabulum, increased obliquity and insufficient coverage of the femoral head by the acetabulum, and increased femoral neck-shaft angles. Radiological features of knee joint were bony spur or bony bridge,osteoporosis,incomplete and thickening articular surface with sclerosis and deformation, degenerative disorders. CONCLUSION: Main clinical manifestation of unknown osteoarthritic disease in Chongzhou city was fixed pain of bilateral knee joint or combined with hip joint pain at early stage.and functional limitation in hip joint at late stage. Main radiological feature was hip joint degeneration. To effectively control the osteoarthritic disease of this area, much more etiology researches should be done. PMID- 17419215 TI - [Electrophysiological study on motor nerve implantation after ectopic transplantation of skeletal muscle in rat]. AB - OBJECTIVE: To study the effect of motor nerve implantation after ectopic transplantation of skeletal muscle on nerve regeneration in rat. METHODS: Sixty Sprague-Dewley male 8-monthold rats were randomly divided into 3 groups: control group, in situ implantation group and ectopic transplantation group. In control group, obturator nerve controlling right gracilis was cut off. In in situ implantation group, the right gracilis was cut off and replanted to its original site, and the obturator nerve was implanted to the muscle. In ectopic transplantation group, the right gracilis was cut off and transplanted to the muscle of the left leg, and the obturator nerve was implanted to the muscle. After 25 weeks, the neurophysiological information was collected through electromyography and the weight of the muscle was measured. RESULTS: The potential without control of the nerve existed in control group. There were no significant differences in latency, amplitude and conduct velocity between in situ implantation group and ectopic transplantation group (P > 0.05). The atrophy of gracilis was dominant in control group, the weight of the muscle was 158.0 +/- 19.3 mg. The weights of the muscle were 509.6 +/- 14.5 mg in ectopic transplantation group and 516.8 +/- 12.7 mg in in situ implantation group, showing no significant difference (P > 0.05). The weights of the muscle in in situ implantation and ectopic transplantation group were larger than that in control group, showing significant difference (P < 0.05). CONCLUSION: Motor nerve implantation after ectopic transplantation of skeletal muscle could prevent the atrophy of the muscle and resume partial function of nerve. PMID- 17419216 TI - [Effect of calcium on reconstruction of basement membrane in composite chitosan tissue engineering skin]. AB - OBJECTIVE: To search the most suitable concentration of calcium in the medium for the basement membrane reconstruction in tissue engineering skin in vitro. METHODS: Composite chitosan tissue engineering skin was prepared according to previous studies. Four groups were included according to the concentration of calcium (1.00, 1.45, 1.65 and 1.95 mmol/L respectively). After 7 days and 15 days of culture, the histological manifestation of basement membrane in tissue engineering skin was observed by hematoxylin & eosin staining and PAS staining, and collagen IV of basement membrane was detected immunohistochemically at the dermal-epidermal junction. RESULTS: This tissue engineering skin shared some histological features of normal skin, including a well-differentiated stratified epidermis and a dense dermis. The epithelium in the group of 1.95 mmol/L. Llcium differentiated better than those in other groups. PAS staining showing a regularly red dying strap domain at the dermal-epidermal junction. Collagen IV was positively stained immunohistochemically at the dermal-epidermal junction in the tissue engineering skin. CONCLUSION: The above results suggest that the medium with 1.95 mmol/L calcium should be suitable for the growth of composite chitosan tissue engineering skin and the reconstruction of basement membrane. PMID- 17419217 TI - [Effects of different stress environments on growth of tissue engineering blood vessels]. AB - OBJECTIVE: To explore the influence of different stress environments on the growth of tissue engineering blood vessels in vivo. METHODS: The engineering vascular scaffolds were prepared with the porcine small intestinal submucosa(SIS) wrapping vascular endothelial cells and smooth muscle cells,which were implanted into the subcutaneous tissue (subcutaneous group), the femoral quadriceps (intramuscular group), and sheathed the femoral artery (perivascular group) respectively. Four weeks postoperatively, these cultured tissues were harvested, and evaluated by macroscopic observation and histology detection. RESULTS: The cultivated tissues in different stress environments had obvious difference in respect of the tubular configuration, cellular proliferation and tissue shape. In subcutaneous group, the wall structure integrity, seed cell proliferation and SIS scaffold decomposition were poor, lumen surface was covered without endothelial cells; in intramuscular group, integrity tubular structure had formed, seed cell proliferation was found to a certain extent, lumen surface was covered with sparse endothelial cells, and a little SIS scaffold was found, cellular and fiber structured arranged irregularly; in perivascular group, vascular-like structure formed, the seed cell growth and proliferation were good, the lumen surface was completely covered with endothelial cells, the smooth muscle cells were in good morphological distribution, the anti-hydrostatic pressure was 247.0 +/- 3.5 kPa, showing significant differences when compared with subcutaneous group (67.0 +/- 5.8 kPa) and intramuscular group (104.0 +/- 7.6 kPa) (P < 0.01). The total scoring of tissue engineering blood vessel formation in subcutaneous group, intramuscular group and perivascular group were 5.529 +/- 0.272, 8.875 +/- 0.248 and 14.824 +/- 0.253 respectively, and the differences among them were significant (P < 0.05). CONCLUSION: Stress excitation has a great influence on the cellular proliferation and the growth of tissue engineering blood vessel in vivo. PMID- 17419218 TI - [Research advance of role of inhibitor of differentiation 2 in skeletal muscle regeneration]. AB - OBJECTIVE: To review researches of the role of inhibitor of differentiation 2(Id2) in skeletal muscle regeneration. METHODS: The latest original literature concerning Id2 and its role in skeletal muscle regeneration was extensively reviewed. RESULTS: Id2 could form heterodimers by combining with E protein to prevent myogenic regulatory factors (MRFs) forming heterodimers by combining with E protein, to inhibit the transcription activity of MRFs and differentiation of skeletal muscle cell. CONCLUSION: Id2 plays an important role in skeletal muscle regeneration. PMID- 17419219 TI - Low- and high-grade non-invasive gastric neoplasia (formerly dysplasia): cytological differentiation (gastro-entero-pancreatic antigens) in association with p53 pattern expression. AB - BACKGROUND/AIMS: In order to better define the evolutive potentiality of non invasive neoplasia (formerly dysplasia) a study of the cytological differentiation and of the behavior of p53 in relation to the clinical progress has been performed. METHODOLOGY: Gastro-entero-pancreatic antigens, p53 and Ki-67 expression were evaluated in 120 cases of epithelial gastric dysplasia: 70 cases of low-grade dysplasia (LGD) and 50 cases of high-grade dysplasia (HGD). For the cytological study four antigens were studied: two of them gastric (pepsinogen C, gastric foveolar M1), one enteric (CAR-5) and one pancreatic (DU-PAN-2). Routinely processed tissue sections of a colon carcinoma known to contain mutant p53 were used as positive controls for p53 immunohistochemistry. For Ki-67 immunohistochemistry, routinely processed tissue sections of normal lymph node and tonsil were used as staining controls. RESULTS: The study of the coexpression of the gastro-entero-pancreatic antigens showed how all cases of non-invasive neoplasia associated with or progressed to gastric carcinoma (GC) were characterized by entero-pancreatic markers and, in particular, in case of LGD p53 expression positivity was evidenced in 66.6% of cases. Ki-67 hyperproliferation is present in 100% of cases. CONCLUSIONS: The cytological study, only if confirmed by wider casuistries, could represent further information in order to better define the follow-up and the therapeutical decisions in case of non invasive gastric neoplasia therefore, the immunophenotypic study in association with p53 could lead to more personalized therapeutical choices. PMID- 17419220 TI - Laparoscopy-assisted distal gastrectomy with 3-cm laparotomy, left hepatic lobe compression technique, and selection of automatic anastomosis device. AB - In this study, we performed laparoscopy-assisted distal gastrectomy (LADG) and lymph node dissection with an incision of 3 cm aiming at radical cure and low invasiveness. We introduce and discuss this technique of minilaparotomy and recommend a device for anastomosis. In LADG, a skin incision of 5cm or greater is made in order to pull out the stomach in other institutes. Whether function is distinctly better after laparoscopy-assisted surgery than after abdominal section has not been elucidated so far, so we should seek an aesthetic advantage. We have used a 3-cm abdominal wound to date. If the wound is smaller than this, the body of the SDH25 cannot be inserted, and currently a wound less than 3cm may thus not be possible. The shaft of the SDH is straight, making it easy to confirm the direction even through a laparoscope. The shaft of the anvil head of the PPCEEA is too long, so that when it is connected with the body through the 3-cm incision, it is necessary to draw it through the remnant stomach to a great extent. PMID- 17419221 TI - A proposal of an appropriate surgical approach for cancer of the ampulla of Vater: retrospective analysis of 73 resected cases. AB - BACKGROUND/AIMS: Because early ampullary cancer has a good prognosis, less invasive surgery should be considered. But recent reports point out limitations of ampullectomy. METHODOLOGY: Between April 1975 and March 2005, seventy-three patients with ampullary cancer were treated. The survival rates of different clinicopathologic features were analyzed retrospectively. RESULTS: Macroscopically, N(-) (negative lymph node metastasis), Panc(-) (no invasion of the pancreatic parenchyma) patients had a significantly longer 5-year survival rate than N(+) (positive lymph node metastasis), Panc(+) (invasion of the pancreatic parenchyma) patients (61.1% vs. 23.1%, 62.2% vs. 21.9%). Histologically, n(-), panc(-), and du(-) (no invasion of the duodenum) patients also had a significantly longer 5-year survival rate than n(+), panc(+), and du(+) (invasion of the duodenum) patients (63.3% vs. 21.1%, 64.3% vs. 29.8%, 83.3% vs. 36.8%, respectively). Patients with Panc(+), Du(+), mixed type and tumors other than the exposed type had significantly more lymph node metastases. CONCLUSIONS: We propose PpPD and regional lymph node dissection as the reasonable operative method. If the tumor is preoperatively diagnosed as Panc(-), Du(-) and N(-), less invasive surgery may be indicated. PMID- 17419222 TI - Three-port versus four-port laparoscopic cholecystectomy. AB - BACKGROUND/AIMS: Since the first successful laparoscopic cholecystectomy with the establishment of pneumoperitoneum in France by Mouret in 1987, it has become the golden standard for cholecystectomy. Generally techniques with four trocars have been used with surgeons but some of them prefer 3-trocar techniques. Our aim is to compare the clinical outcomes of three- and four-port techniques prospectively. METHODOLOGY: Between 1998 and 2003, one hundred and forty-six consecutive patients who underwent elective laparoscopic cholecystectomy for cholelithiasis in the Medical Faculty of Suleyman Demirel University were randomized to receive either the three-port or the four-port technique. Operative time, (time from the beginning of the insufflation up to the closure of the skin), success rate, visual analogue pain score, analgesia requirements, postoperative hospital stay were compared. RESULTS: No differences between the two groups could be found. CONCLUSIONS: Three-port technique is safe, effective, and economic but does not reduce the overall pain score and analgesia requirement. PMID- 17419223 TI - Fascin expression is correlated with tumor progression of extrahepatic bile duct cancer. AB - BACKGROUND/AIMS: Fascin, an actin-crosslinking protein, participates in cell motility. Fascin over-expression induces a high potential for invasion and metastasis in various malignancies. The aim of this study was to determine the relationship of fascin expression to clinicopathological findings in patients with extrahepatic bile duct cancer. Furthermore, we investigated the correlation between fascin expression and intracellular adhesion molecular (E-cadherin and beta-catenin). METHODOLOGY: We evaluated the expression of fascin, E-cadherin and beta-catenin by immunohistochemistry in surgical specimens from 26 patients with extrahepatic bile duct cancer. RESULTS: Normal epithelial cells of the bile duct was not immunoreactive for fascin, and cancer cells often show immunoreactivity, which was found more frequently at the invasive tumor fronts than at other tumor areas. The present study demonstrated a statistically significant correlation between fascin expression and gender, tumor status, vascular invasion, and disease stage. We detected that increased immunoreactivity for fascin had tendencies to disrupt membranous immunoreactivity for E-cadherin and beta catenin. CONCLUSIONS: We conclude that fascin expression is correlated with tumor progression. The expression of fascin is frequently detected at the invasive tumor fronts, indicating that invading tumor cells express fascin abundantly. In tumor cells with an over-expression of fascin, E-cadherin and betacatenin expressions often disrupt membranous immunoreactivity. PMID- 17419224 TI - Risk factors for infectious morbidity in gallbladder cancer patients treated surgically. AB - BACKGROUND/AIMS: To explore risk factors associated with postoperative infectious morbidity in gallbladder cancer patients. METHODOLOGY: We investigated 58 consecutive patients undergoing surgery for primary gallbladder cancer between January 2000 and June 2005 in our hospital. Records of all patients were retrospectively reviewed. Twenty-two independent tumor-, patient-, and treatment related variables were analyzed. The dependent variable was clinical infectious complications. A binary logistic regression analysis was used to assess the independent association of variables with the dependent variable. RESULTS: Overall surgical morbidity was 33% (19/58), and 14 (24%) of the 58 patients developed infectious complications postoperatively. On univariate analysis, presence of jaundice, hypoalbuminemia and intraoperative blood transfusion were found to be significantly associated with infectious morbidity. The multivariate analysis of logistic regression disclosed that presence of jaundice and intraoperative blood transfusion of 4 units or more only showed marginally significant impacts on infectious complications (odds ratio, 8.004, 7.782; 95% confidence interval, 0.886-72.278, 0.914-66.283, respectively), while weight loss and hypoalbuminemia were significantly associated with infectious complications postoperatively (odds ratio, 9.609, 40.257; 95% confidence interval, 1.269 72.253, 3.901-415.438, respectively). CONCLUSIONS: Hypoalbuminemia and weight loss are significantly associated with postoperative infectious morbidity independently. While presence of jaundice and intraoperative blood transfusion of 4 units or more appear to be marginally significant factors, modality of operation or liver resection, blood loss, and additional gastrointestinal operation are not risk factors. PMID- 17419225 TI - Management of bile leakage caused by clip displacement from cystic duct stumps. AB - BACKGROUND/AIMS: Cystic duct leakage (CDL) is one of the most common complications after laparoscopic cholecystectomy (LC). We retrospectively examined 3 patients with CDL that were treated with endoscopic stenting. METHODOLOGY: From January 1995 to December 2004, 1127 LCs were performed. CDL occurred in 3 cases and a closed suction drain was inserted in 2 of those 3 cases. RESULTS: The duration from LC to endoscopic retrograde cholangiopancreatography (ERCP) was 2 to 4 days, and the patient without the drain experienced the longest waiting time. Hospital stay was almost identical for all 3 cases. There was an immediate improvement after the procedure. CONCLUSIONS: It is important to employ accurate technical surgical procedures to prevent complications. ERCP with stent placement and sphincterotomy is an easy and safe diagnostic and therapeutic procedure for CDL, and has a high success rate in resolving leaks. PMID- 17419226 TI - Inhibition of subgenomic hepatitis C virus RNA replication in HeLa cells. AB - BACKGROUND/AIMS: Antiviral therapy such as combination interferon and ribavirin can eradicate hepatitis C virus (HCV) RNA by up to 40-50%. However, many patients still remain non-responders to this treatment for various reasons. The aim of this study was to evaluate the effect of interferon or ribavirin treatment on subgenomic HCV RNA replication in 'non-hepatic' HeLa cells. METHODOLOGY: Huh-7 or HeLa cells harboring HCV replicon were constructed by using cellular RNA of Huh-7 harboring HCV replicon RNAs, named as C13-3 cells. We also tested whether interferon or ribavirin can suppress HCV RNA in HeLa cells. RESULTS: Huh-7 or HeLa cells harboring HCV replicon RNAs were constructed by using cellular RNA of C13-3 cells than using in vitro-transcribed RNA. Ribavirin at 1 microg/mL or 10 microg/mL did not suppress colony formation in HeLa cells, but at 100 microg/mL suppression was observed. Interferon-alpha 2b suppressed HCV replication even at 1 U/mL. CONCLUSIONS: HeLa cells harboring HCV replicon RNAs also might be useful for the development of antiviral drugs. PMID- 17419227 TI - Clinicopathological significance of aminopeptidase N/CD13 expression in human gastric carcinoma. AB - BACKGROUND/AIMS: Aminopeptidase N (APN)/CD13 is a transmembrane ectoenzyme occurring in a wide variety of cells. Recently, APN/CD13 has been reported to be involved in tumor invasion and metastasis. However, precise functions in tumor cells, and its role in gastric carcinoma remain unclear. METHODOLOGY: To evaluate the role of APN/CD13 in gastric carcinoma, we conducted immunohistochemical staining for APN/CD13 in 121 gastric carcinoma specimens, using anti-APN monoclonal antibody. The relationship between APN/CD13 expression and various prognostic factors of gastric carcinoma were investigated. RESULTS: Of the 121 patients with gastric carcinoma studied, 48 were strongly positive, 36 were weakly positive, and 37 were negative. Overall survival rate of the patients with negative APN/CD13 expression was significantly lower than that of the patients with positive APN/CD13 expression. APN/CD13 expression was negatively associated with lymph node metastasis. Multivariate analysis showed APN/CD13 expression to be a significant prognostic factor. CONCLUSIONS: Decreased expression of APN/CD13 was associated with a poor prognosis. Hence, our results demonstrate that the immunohistochemical detection of APN/CD13 could provide useful information as one of the prognostic factors in gastric cancer. PMID- 17419228 TI - Transcatheter arterial coil embolization of iatrogenic pseudoaneurysms after hepatobiliary and pancreatic interventions. AB - BACKGROUND/AIMS: Visceral and renal arteries pseudoaneurysms are uncommon but potentially lethal complications of hepatic and pancreatobiliary interventions. To evaluate the clinical outcome of transcatheter arterial coils embolotherapy, we reviewed our institution's experience with the management for bleeding pseudoaneurysms. METHODOLOGY: From January 1988 through December 2004, 20 patients were encountered who developed massive bleeding from pseudoaneurysms following hepatobiliary and pancreatic interventions. All patients underwent diagnostic angiography and transarterial embolization was carried out thereafter. RESULTS: Embolization was technically successful without major post-procedural complications in all patients. Bleeding was stopped after embolization in 17 patients (85%), and rebleeding did occur in one patient during the follow-up periods. Repeat coil embolotherapy was performed in one patient with recurrent bleeding, but they needed surgical intervention because of failed re embolization. Another two patients needed surgical ligation and one of the patients died of sepsis two weeks later. CONCLUSIONS: An emergency angiography should be considered in all patients in whom pseudoaneurysm is suspected following hepatobiliary and pancreatic interventions. Transcatheter arterial coil embolization is a safe and effective treatment for pseudoaneurysm. Surgical intervention should be reserved for patients for whom embolization fails or for whom it is not possible. PMID- 17419229 TI - Intraperitoneal hyperthermia in the management of pseudomyxoma peritonei. AB - BACKGROUND/AIMS: Pseudomyxoma peritonei is a rare disease characterized by diffuse intraperitoneal mucinous tumor and massive mucinous ascites. The mainstay of treatment is surgery in combination with adjuvant therapy. METHODOLOGY: From 1995 to 2002, 8 patients with pathologically confirmed pseudomyxoma peritonei were studied. After surgical debulking, intraoperative intraperitoneal hyperthermia was performed followed by 5 days of postoperative intraperitoneal hyperthermia. Normal saline at 46 C was used for hyperthermia. Patients were followed for clinical data, survival, morbidity, and mortality. RESULTS: The appendix was the origin of pseudomyxoma peritonei in 6 patients, the ovary in 1 patient, and the fallopian tube in another. The pathologic diagnosis was mucinous adenocarcinoma in 4 patients, mucinous cystadenocarcinoma in 3, and mucinous cystadenoma in 1. Associated morbidity and mortality was 12.5% and 0, respectively. The recurrence rate was 62.5%, with a median follow-up of 52.5 months. The estimated 1-, 3-, and 5-year survival rates were 100%, 88%, and 49%, respectively. CONCLUSIONS: Surgical debulking followed by 46 C normal saline intraperitoneal hyperthermia resulted in a 5-year survival of 49% with low morbidity and no mortality for the treatment of pseudomyxoma peritonei. We concluded that this is an easy, safe, and efficacious treatment for patients with this rare disease. PMID- 17419230 TI - Sialoglycoconjugate expression in primary colorectal cancer and metastatic lymph node tissues. AB - BACKGROUND/AIMS: Aberrant cell surface glycosylation, and especially excessive sialylation, is thought to have great importance in tumor malignancy. To investigate the clinicopathological significance of sialylation in colorectal cancer, we performed histochemical analyses using sialic acid-binding lectins. METHODOLOGY: Primary colorectal cancer and lymph node tissues obtained from 80 cases were subjected to lectin-immunohistochemical staining using Maackia amurensis leukoagglutinin (MAL) and Sambucus nigra agglutinin (SNA). The relationship between the staining characteristics and the various clinicopathological parameters was statistically analyzed. RESULTS: In primary cancer tissues, a high level of SNA staining was significantly related to worse prognosis and some pathological characteristics such as lymph node metastasis, whereas a high level of MAL staining was related to worse prognosis. In metastatic lymph node tissues, positive staining was frequently found for MAL and SNA, which wasremarkable in cases categorized as N2 metastasis. Furthermore, cases with MAL-positive staining in metastatic lymph node tissues evidently indicated worse prognosis than those with MAL-negative staining. CONCLUSIONS: Aberrant expression of SNA-positive sialoglycoconjugates in primary colorectal cancer tissues is important in terms of unfavorable pathological characteristics of the cancer. In addition, aberrant expression of MAL-positive sialoglycoconjugates in metastatic lymph node tissues might be related to worse prognosis. PMID- 17419231 TI - Defecation states in patients with or without soiling at 5 years or more after colectomy, mucosal proctectomy and ileal J pouch-anal anastomosis for ulcerative colitis. AB - BACKGROUND/AIMS: There are no reports about the relationship between soiling and other defecation states in patients with ulcerative colitis (UC) after ileal J pouch-anal anastomosis (IPAA). To assess the defecation states in patients with or without soiling at 5 years or more after IPAA for UC, we studied clinical findings in such patients. METHODOLOGY: Subjects with UC who had undergone IPAA [a 5-cm short cuff, anal canal mucosa including anal transitional zone (AZT) stripped from the level of dentate line (DL)] with ileostomy closure at least 60 to 132 months (mean; 103.6 months) previously, and who had no pre- or postoperative complications were recruited. They were 43 patients (28 men and 15 women aged 15 to 59 years, average 39.2 years) divided into 2 groups; group A n=28; patients without soiling (18 men and 10 women aged 15 to 49 years, average 37.5 years) and group B n=15; patients with soiling (10 men and 5 women aged 24 to 59 years, average 42.1 years). Postoperative interviews concerning the defecation states (mean daily stool frequency, nocturnal stool frequency, ability to discriminate flatus from feces, feeling of stool remaining, consistence of stools, soiling, incontinence, and urgency) were conducted with patients after IPAA. RESULTS: No cases in group A showed soiling. All cases in group B had some soiling, 6 cases; rare soiling (at diarrhea), 4 cases; occasional soiling (1 time per 2 or 3 days), 4 cases; frequent soiling (patients must use pad due to daily soiling, 2 cases had incontinence). There was a significant difference between groups A and B (P < 0.001). All cases in group A showed less than 6 times per day and all cases of group B showed 7 or more times per day. There was a significant difference between groups A and B (P < 0.001). High nocturnal stool frequency was significantly more common in patients of group B than group A (P < 0.001). All cases in group A and 12 cases in group B could significantly discriminate flatus from feces (P < 0.05). Feeling of stool remaining was significantly more common in patients of group B than group A (P < 0.001). The stool consistency of group A was significantly harder than that of group B (P < 0.001). No cases in group A reported incontinence. Two cases in group B had incontinence. Urgency in group B was noted more than in group A. Regarding patient satisfaction (Peck's criteria), all cases in group B and no cases in group A reported "excellent". Ten cases in group B were "good". Three cases in group B were "fair". Two cases in group B showed failure. Patient satisfaction of group A was significantly higher than that of group B (P < 0.01). CONCLUSIONS: According to these interviews, no importance of preserving the AZT was recognized. The patients without soiling showed better defecation states than patients with soiling. The patients with soiling showed poor defecation status following PMID- 17419232 TI - Role of regular follow-up after curative surgery for colorectal cancer. AB - BACKGROUND/AIMS: This investigation aimed to estimate the value of regular follow up programs after curative resection for colorectal cancer. METHODOLOGY: We compared the recurrence rate, rate of curative re-resection, and survival rate between groups who had either regular or irregular follow-up. The medical records of 397 consecutive patients, who underwent a curative resection for colorectal cancer between January 1994 and December 1997, were analyzed retrospectively. RESULTS: The recurrence rate was 19.4% and 20.8% in the regular and irregular follow-up groups (P > 0.05), respectively. There was a significant difference in the asymptomatic recurrence rate (62.9 vs. 18.7%; P = 0.021), but curative re resection was possible in 18 (29.0%) of those patients with cancer recurrence in the regular follow-up group, and in 2 (12.5%) in the irregular follow-up group, which was not significantly different (P > 0.05). 5-year survival rate between the groups was not significantly different (78.1 vs. 61.2%; P > 0.05). CONCLUSIONS: A regular follow-up program after a curative resection for colorectal cancer, although facilitating detection of recurrence before symptoms developed, was unlikely to succeed in increasing the rate of a curative intent re resection and survival remarkably. PMID- 17419233 TI - Pit pattern type analyses of colon polyps with high-resolution colonoscope. AB - BACKGROUND/AIMS: Correlations among pit pattern types and some characteristics of colon polyps were assessed with high-resolution colonoscope in Turkish patients. METHODOLOGY: Sixty-five patients were included in the study. All visible polyps and distal 30cm of colon were stained with indigo carmine 0.4% after standard colonoscopic examination. Then, pit pattern analyses were done. Correlations were evaluated and results were interpreted as significant if p < 0.05. RESULTS: There were correlations among the pit pattern types of polyps and their size, location, morphology and histology. Adenomatous polyps (type III,IV,V) were mostly located in the right colon, nonadenomatous polyps (type I/II) were mostly in the left. Ratio of having adenomatous structure increased in a parallel course with increase in polyp's size. While most of the nonadenomatous polyps were with type I,II, adenomatous polyps were with type III, IV, V patterns. Most of the type II, III, IV polyps were sessile and type I polyps were flat. Sensitivity and specificity of adenomatous and nonadenomatous polyps were 80% and 89% respectively and overall accuracy rate was 87%. Type III/IV were the best estimated type among the others. After chromoendoscopy, 35% increase in polyp number was also detected. CONCLUSIONS: High-resolution endoscope is successful for prediction of histology of colorectal polyps. PMID- 17419234 TI - Vascular endothelial growth factor-C (VEGF-C) is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer. AB - BACKGROUND/AIMS: It is useful to decide whether lymphatic involvement or lymph node metastasis exists before polypectomy or operation in submucosal colorectal cancer. Whether vascular endothelial growth factor-C (VEGF-C) or VEGF-D could predict lymph node metastasis and lymphatic involvement is uncertain. METHODOLOGY: Expression of the VEGF-C and VEGF-D in human submucosal colorectal cancers was investigated in paraffin-embedded stepwise sections by means of immunohistochemistry, and the correlation between immunohistochemical expression pattern and clinicopathological features was also evaluated. RESULTS: The results showed that VEGF-C overexpression correlated with lymphatic involvement (P = 0.01) and lymph node metastasis (P = 0.02), but VEGF-D overexpression did not correlate significantly. In multivariate analysis lymphatic invasion was the predictive factor (P = 0.0129), but VEGF-C positivity was not predictive (P = 0.3437). CONCLUSIONS: These results may suggest that VEGF-C is a more specific risk factor for lymph node metastasis than VEGF-D in submucosal colorectal cancer. PMID- 17419235 TI - Subsite distribution of colorectal carcinoma and implications for screening; a retrospective audit of 1771 cases. AB - BACKGROUND/AIMS: Recent epidemiologic studies mentioned a shift from left to right in colorectal cancer. We searched our hospital database to try to give an insight into the patient characteristics and also optimize the screening programs in our country. METHODOLOGY: A total of 1771 colorectal cancer patients were identified and analyzed. Patients' gender, age, operation dates, tumor localization, resectability and T stage of the lesions, were recorded and comparisons were made. Second step of the study consisted of a questionnaire including 6 questions that were asked to 278 physicians to document current physician attitudes and practices regarding colorectal cancer screening, and to formulate appropriate interventions. RESULTS: Most of our patients were older than 51 in our study and a higher percentage of cases had sigmoid and rectal cancer (67%). The chronological trends in anatomic subsite distribution of colorectal cancer showed that by changing years when left colon cancer percentage was increasing, rectum cancer percentage was decreasing. There was no age dependent variation in the topographical distribution of colorectal cancer by changing years. Also there was no relation between sex differences and subsite distribution when the whole group was taken into consideration. Specialist physicians had a poor understanding of colorectal cancer screening and lacked the responsibility of the screening programs. CONCLUSIONS: Data presented in this article do not support a progressive left to right shift in the distribution of colorectal carcinoma. Screening with fecal occult blood testing and sigmoidoscopy seems appropriate. Physician education may be an important step in screening before public awareness. PMID- 17419236 TI - Clinicopathological features of skip metastasis in colorectal cancer. AB - BACKGROUND/AIMS: Japanese general rules for the staging of colorectal cancer conventionally classify lymph node metastasis into three groups according to location with respect to the primary tumor. Skip metastasis, in which distant nodes are positive but regional nodes are negative, is often encountered but poorly understood. We studied the clinicopathological features of skip metastasis in colorectal cancer. METHODOLOGY: The location of positive nodes was classified in 323 patients with Dukes' stage C colorectal cancer. Skip n2 lymph node metastasis was defined as positive N2 metastasis without negative N1 or N3 metastasis. Clinicopathological findings and survival were compared between the patients with skip n2 metastasis (skip n2 group) and those with n1 (n1 group) or n2 metastasis (n2 group). RESULTS: There were 211 patients in the n1 group, 91 in the n2 group, and 21 in the skip n2 group. Pathological examination showed that the skip n2 group had fewer positive nodes than the n1 and n2 groups, but was positioned between these groups with respect to the degree of lymphatic invasion. Cumulative survival was significantly poorer in the n2 group than in the skip n2 group (p = 0.039 by log-rank test). Survival was similar in the skip n2 group and n1 group. There was also no difference in survival between patients in the skip n2 group and patients with one, two, or three N1 metastases. CONCLUSIONS: Lymph nodes with skip n2 metastasis are most likely sentinel nodes of the primary tumor in patients with colorectal cancer. The prognosis of patients with skip n2 metastasis is therefore better than that of patients with n2 metastasis and similar to that of patients with n1 metastasis. PMID- 17419237 TI - Laparoscopic mesorectal excision with preservation of the pelvic autonomic nerves for rectal cancer. AB - BACKGROUND/AIMS: Laparoscopic mesorectal excision with preservation of the autonomic pelvic nerves for rectal cancer including selected advanced lower rectal cancer is now challenging. The aims of the study were to assess the surgical results and short-term outcomes of this procedure prospectively. METHODOLOGY: Seventy-four of 281 rectal cancer patients, since the introduction of laparoscopic colorectal surgery in our hospital, have undergone laparoscopic rectal surgery. The location of the tumor distributed in upper rectum; 33, middle; 22, and lower 19. The mesorectal excision with preservation of the autonomic pelvic nerves was performed for all the patients. The laparoscopic mesorectal excision was performed under 8 to 10 cmH2O CO2 pneumoperitoneum and lymph node dissection was performed along the feeding artery depend on individuals. Ipsilateral lateral lymph node dissection was added for 5 cases of advanced lower rectal cancer. RESULTS: Open conversion occurred in 4 cases, 2 of those were due to locally advanced tumors and 2 technical difficulties in transaction of the distal rectum. There were 15 postoperative complications, 7 anastomotic leakage (10.6%), 3 transient urinary retention (4.1%), 4 wound infection (5.3%), and 1 small bowel obstruction (1.4%). No mortality was recorded in this series. Time of operation was 203 +/- 54 min in mesorectal excision cases and 270 +/- 42 min mesorectal excision with lateral lymph node dissection cases. Blood loss was 92 +/- 90g and 276 +/- 66 g respectively. The hospital length-of stay was 11.7 days in average. CONCLUSIONS: Laparoscopic mesorectal excision with preservation of autonomic pelvic nerves for rectal cancer patients including selected advanced lower rectal cancer is favorable. PMID- 17419238 TI - Assessments of anorectal myectomy for short segment hypoganglionosis in childhood. AB - BACKGROUND/AIMS: To clarify the significance of anorectal myectomy (ARM) in childhood patients with short segment hypoganglionosis (Hypo), we analyzed the clinical features of these patients before and after ARM. METHODOLOGY: A consecutive series of twenty-nine patients with short segment Hypo were included in this study. These were sixteen males and thirteen females, aged between 6 and 15 years with a mean age of 9.8 years. We performed an analysis of the clinical findings and outcomes for patients with short segment Hypo before and after ARM. Stool frequencies were less than twice per week (0.5-2 per week, mean: 1.6 per week) despite the use of laxatives, suppositories, and enema before ARM. RESULTS: About 90% of patients had an onset of constipation before the age of 6 years. In addition, significant differences were noted between 0 months < or = to <1 year and 1 year < or = to < or =15 years (P < 0.0001). There were no sexual differences. Difficulty in defecation was the most common symptom, followed by abdominal fullness, abdominal pain and soiling. In abdominal fullness and pain, there were significant differences between positive and negative groups (P < 0.0001, P = 0.0038, respectively). No patients responded to use of laxatives, suppositories, and enema before ARM. In patients after ARM, the most common outcome was good (65.5%; bowel movement every day without laxatives and no other symptoms), followed by satisfactory (20.7%; bowel movement 2-3 times per week without laxatives and no other symptoms) and poor (13.8%; no improvement) in decreasing order. Significant differences of patient numbers were noted between good and the remaining outcomes (P = 0.0181). Spontaneous evacuation without the need for laxatives, suppositories, and enema was recorded in 100% patients of good outcome, and 42.9% those of satisfactory outcome after ARM. Patients with poor outcome still required for laxatives, suppositories, and enema. Central nervous, psychological, and psychiatric disorders were recognized in patients with satisfactory outcome, and constipation was maintained to these patients. However, the necessity for medication to them was lost. Moreover, clinical symptoms disappeared. Redundant colon was only found in patients with poor outcome. CONCLUSIONS: Patients with short segment Hypo, especially those with redundant colon, psychological, psychiatric, or central nervous disorders, may experience persistent constipation after ARM. PMID- 17419239 TI - Values of Doppler sonography predicts high risk variceal bleeding in patients with viral cirrhosis. AB - BACKGROUND/AIMS: Gastrointestinal bleeding such as rupture of esophagogastric varices remains one of the leading causes of death in patients with liver cirrhosis. As a critical issue, assessment of the bleeding risk of esophageal varices is extremely important. In the present study, by determining the relationship between several parameters measured by pulsed Doppler sonography and the bleeding risk of esophageal varices assessed by upper endoscopy, we investigated what is the most valuable parameter as a supplement to the bleeding risk. METHODOLOGY: A total of 158 patients with hepatitis virus-infected liver cirrhosis (56 positive for HBs antigen and 102 positive for HCV antibody) were studied. As controls, 171 normal subjects were used. The flow volumes of the portal trunk and the splenic vein, the Congestion Index, and the S/P ratio were measured by pulsed Doppler sonography. Based on upper endoscopic findings, we classified the patients into two groups based on bleeding risk of esophageal varices: high-risk and low-risk. Logistic regression analysis was employed to identify the most valuable parameter as a supplement to the bleeding risk. RESULTS: The flow volume of the splenic vein, the Congestion Index, and the S/P ratio in cirrhotic patients with esophageal varices were significantly higher than those in normal subjects (P = 0.000). The mean flow volumes of the portal trunk and splenic vein and the mean of the S/P ratio in the high-risk group for bleeding of esophageal varices were significantly higher than those in the low risk group (P = 0.000-0.005). Based on logistic regression analysis, the flow volume of the splenic vein was found to be the most valuable parameter for bleeding risk (P < 0.001). CONCLUSIONS: The flow volume of splenic vein with pulsed Doppler sonography was the most valuable parameter for the bleeding risk of esophageal varices. PMID- 17419240 TI - Surgical outcome of patients with thoracic esophageal cancer positive for cervical lymph nodes. AB - BACKGROUND/AIMS: Although cervical lymph nodes were classified as distant metastases in patients with thoracic esophageal cancer, not a few patients survive more than five-years. The purpose of this study was to predict patients with good prognosis among thoracic esophageal cancer patients with cervical node metastases. METHODOLOGY: From 1983 to 2002, 312 consecutive patients with thoracic esophageal squamous cell carcinoma underwent curative surgery with 3 field lymph node dissection (3FLD). A total of 88 (28%) of 312 patients were diagnosed with cervical lymph node metastases. Univariate and multivariate analyses were carried out to evaluate the impact of clinico-pathological factors on the survival of these patients. RESULTS: Overall five-year survival rate of 88 patients with cervical lymph node metastases was 26%. Univariate analysis revealed that following groups showed more than 40% overall five-years survival rate; female patients, patients with T1, T2 tumors and patients without thoracic node metastases. These variables were also independent good prognostic factors in multivariate analysis. CONCLUSIONS: Although cervical lymph node metastases was risk factors for worse survival, female patients, patients with T1, T2 tumors and patients without thoracic node metastases showed acceptable overall survival after 3FLD. PMID- 17419241 TI - Comprehensive investigations of quality of life after esophagectomy with special reference to the route of reconstruction. AB - BACKGROUND/AIMS: Esophagectomy is a very invasive operation, therefore, it is important to improve the postoperative quality of life (QOL) of the patients. The aim of this study was to evaluate the QOL of patients who had undergone esophagectomy for thoracic esophageal cancer. METHODOLOGY: We investigated 37 patients who had undergone esophagectomy. The anastomosis was made at the cervical location by the retrosternal route in 12 patients (RS group), at the high thoracic location by the posterior mediastinal route in 18 patients (HT group), and at the cervical location by the posterior mediastinal route in seven patients (PM group). QOL was evaluated by patient questionnaires concerning reflux esophagitis using QUEST and dumping syndrome, body weight, ambulatory pH monitoring, and immunostaining for iNOS and COX-2 as markers of inflammation. RESULTS: The QUEST score revealed that the findings suggesting reflux were few in the HT group. Patients suffered from dumping syndrome were significantly few in the HT group (p = 0.0399). The percentage time of pH < or =4.0 was shortest in the HT group at the position of the esophagogastric anastomosis (p < 0.0281). Body weight recovery was best in HT group (p < 0.0001). There was a tendency that iNOS and COX-2 immunoreactivity were weaker in HT group than other two groups. CONCLUSIONS: Our results suggest that QOL after esophageal reconstruction using a gastric tube is good in patients with the anastomosis at the high thoracic location by the posterior mediastinal route. PMID- 17419242 TI - Relationship between cytokine response and temporary ventilation during one-lung ventilation in esophagectomy. AB - BACKGROUND/AIMS: Postoperative pulmonary complications are often fatal in patients with esophageal cancer. The aim of this study was to clarify the association between cytokines and temporary ventilation during one-lung ventilation. METHODOLOGY: Serum and BALF IL-6, 8 were measured in 39 patients with esophageal cancer, and the relationship between cytokine response and temporary ventilation during one-lung ventilation in esophagectomy was examined. RESULTS: Pulmonary complications did not occur in our patients. Serum IL-6 levels were significantly increased in the high one-lung ventilation ratio and long-term one-lung ventilation groups. Serum IL-6 levels in the one or more times ventilation group during one-lung ventilation were significantly low compared to the no-ventilation group. There were no significant differences in serum IL-8 and BALF IL-6, 8 levels. CONCLUSIONS: Our results indicate that it is important to ventilate one or more times during one-lung ventilation in order to reduce postoperative pulmonary complications. PMID- 17419243 TI - Anti-reflux pouch-esophagostomy after proximal gastrectomy with jejunal pouch interposition reconstruction. AB - Reflux esophagitis is a serious postoperative complication for patients undergoing gastrectomy. We designed a new jejunal pouch-esophagostomy to prevent reflux after proximal gastrectomy. After proximal gastrectomy, ajejunal segment about 17 cm long was folded. Side-to-side jejuno-jejunostomy was made using a linear stapler with 100-mm staples along the length at the anti-mesenteric side. A 10-cm-longjejunal pouch with a 7-cm-long apical bridge was made. Esophago jejuno end-to-side anastomosis (pouch-esophagostomy) was made with circular stapler at the right anterior wall the apical bridge. We add "partial posterior fundoplication" like wrapping using the apical bridge of the jejunal pouch. Patients with this new anti-reflux anastomosis showed no reflux on barium meal study even in the right anterior oblique deep Trendelenburg's position. Jejunal pouch reconstruction with partial posterior wrapping provides a satisfactory result with regard to preventing reflux esophagitis. PMID- 17419244 TI - Evaluation of the two immunochromatographic methods for detecting urine and serum IgG antibodies to Helicobacter pylori and comparison of accuracy and clinical utility. AB - BACKGROUND/AIMS: To evaluate the accuracy and clinical utility of two immunochromatographic methods, the STAT-PAK and RAPIRUN tests, in detecting H. pylori antibodies in serum and in urine separately. METHODOLOGY: 130 patients undergoing gastroendoscopy (70 men and 60 women; mean age, 50.6 years) were enrolled in this study. Their H. pylori status was determined based on 4 tests: CLO test, culture, histology and UBT test. The H. pylori positive status was confirmed when the culture was positive, or 2 of the other 3 tests were positive. Serum samples and urine samples were collected. We used the STAT-PAK test for detecting H. pylori antibodies in serum, and read the results 24 hours later. Antibodies were also detected in urine by the RAPIRUN test, with a reading time of 25 minutes. 3 doctors read the results of both tests separately, and a positive result was determined when at least 2 of the doctors read a positive result. RESULTS: In the STAT-PAK pretest using serum samples from 21 randomly selected patients (16 patients with H. pylori infection and 5 patients without infection), the sensitivity at 1-hour and 24-hour reading time was 18.7% and 37.5%. Specificity in both reading times was the same at 100%. Of 129 patients, 82 (64%) were H. pylori infected and 47 (36%) were uninfected. The sensitivity, specificity, and positive and negative predictive values were 31.7%, 93.6%, 89.6% and 44.0% in the STAT-PAK test with a 24-hour reading time, and 93.9%, 85.1%, 91.6%, 88.8% in the RAPIRUN test. One female was excluded because of an invalid urine test. CONCLUSIONS: The urine RAPIRUN test is a faster and more accurate office-based test than the serum STAT-PAK test for detecting the H. pylori infection in untreated patients in Taiwan. The non-invasive urine RAPIRUN test can be used as a large scale screening test for H. pylori status, particularly in children, uncooperative patients, those who have a family history of gastric cancer, or patients who can not tolerate the suffering of endoscopic examination. PMID- 17419245 TI - Prognosis of gastrointestinal stromal tumors. AB - BACKGROUND/AIMS: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors (GIMT) of the gut. The aim of this retrospective study is to correlate the histological risk factors with the survival of our patients operated for GIST. METHODOLOGY: In our department, from 1980 to October 2003, 15 patients were operated for GIST. Their mean age was 58 years old and 8 of them were males; 10 (67%) were localized in the stomach and 5 (33%) in the small bowel. In 7 cases liver metastases were present at laparotomy and 4 of them also had peritoneal diffusion. We performed immunohistochemistry for c-Kit, SMA and S100p. Mitotic index (MI) and size neoplasm were the main pathological criteria for malignity. The patients with c-kit (CD117) positive neoplasms were divided according to NIH Consensus Conference risk class, MI, tumor size, localization, SMA or S100p presence, liver metastasis and peritoneal metastasis to compare the different 5-year survival rates. Survival analysis was performed using Kaplan Meier method and log-rank test and a p < 0.05 was considered as significant. RESULTS: Global survival rate after 5 years was 40% and the mortality was, in all cases, due to GIST. In our experience gender, age, tumor size, localization and S100p positivity did not play any role in predicting the prognosis of GIST. On the contrary high MI and SMA positivity are significantly associated to a lower survival rate (33% vs. 86% and 39% vs. 100% at 5 years, respectively). Finally patients with metastases at laparotomy have a significantly lower 5-year survival rate (hepatic 29% vs. 100%, hepatic and peritoneal 25% us. 78%). CONCLUSIONS: In our experience high MI and in some cases SMA expression can be considered assessed risk factors. On the other hand, criteria of benign behavior did not completely predict the long-term clinical outcome. PMID- 17419246 TI - Small bowel cancer: single-centre results over a period of 12 years. AB - BACKGROUND/AIMS: Tumors of the small bowel are rare, accounting for about 3-6% of all gastrointestinal neoplasms. However, diagnosis and treatment are difficult and an ongoing challenge. METHODOLOGY: Follow-up and clinical data of 43 patients with small bowel cancer who underwent surgery at our hospital. RESULTS: Subgroups consisted of adenocarcinoma (n=16; 37.2%), neuroendocrine tumors (n=12; 27.9%), gastrointestinal stroma tumor (GIST) (n=10; 23.3%), lymphoma (n=3; 7%) and desmoid tumor (n=2; 4.6%). Tumor localizations were within duodenum (46.5%), jejunum (16.3%) and ileum (37.2%). Thirty patients were curatively operated, 13 for palliative treatment or diagnostic purpose. Adenocarcinoma patients showed preponderance of advanced tumor stages: stage I/II in 5 pts, III/IV in 11 patients. Stage distribution for patients with neuroendocrine tumors was 3 each for I and II and 6 for III. Localization was predominantly within the ileum (n=7). Overall survival after five/ten years was 48/37%. Patients with neuroendocrine tumors showed best survival results (75/57%), GIST patients 60/35% and adenocarcinoma (27% each). There was a strong trend towards better survival at early tumor stages in patients with adenocarcinoma and neuroendocrine tumors. CONCLUSIONS: Early diagnosis is essential for prognosis of small bowel malignancies. Cure is unlikely if lymph node or distant metastases have already developed. PMID- 17419247 TI - Clinicopathologic features of Crohn's disease with free intestinal perforation. AB - BACKGROUND/AIMS: Free perforation is the most serious intestinal complication of Crohn's disease, and early diagnosis is an important determinant of survival. The aim of this study was to evaluate the clinical and histopathologic characteristics of free perforation associated with Crohn's disease. METHODOLOGY: Twelve free perforations in 10 patients with Crohn's disease were studied. We mainly examined the clinical characteristics, methods for definitive diagnosis and histopathologic features of free intestinal perforation with Crohn's disease. RESULTS: Nine patients had 1 perforation, and 1 had 3 perforations occurring within a short period. Computed tomography was significantly more sensitive than plain radiography for detecting free air in the abdomen at the time of perforation. Histopathologic examination showed that free perforation occurred most frequently in the ileum, on the side attached to the mesentery. Intestinal strictures and dilatations were presented distal to 10 perforations (83%). CONCLUSIONS: Our results suggest that computed tomography of the abdomen was useful for the definitive diagnosis of free intestinal perforation in Crohn's disease. Histopathologic findings indicated that increased intraluminal pressure due to stricture and local circulatory disturbance of the mucosa caused by intestinal dilatation were involved in the development of free perforation in patients with Crohn's disease. PMID- 17419248 TI - Significance of p53 expression in gastrointestinal stromal tumors. AB - BACKGROUND/AIMS: Gastrointestinal stromal tumors are non-epithelial tumors originating from the gastrointestinal tract walls. They expose a wide spectrum of histological properties from benign to poorly differentiated malignant tumors. METHODOLOGY: In this study 41 cases of gastrointestinal stromal tumors were reviewed for p53 expression using immunohistochemical stains. Immunoreactivity properties of the tumors were compared to prognostic factors such as mitotic index, tumor size, tumor site, presence of necrosis, mucosal invasion, proliferative index, telomerase activity, hemorrhage, cellularity, and histological pattern. RESULTS: There was a significant difference in positive immunostaining of three case groups (12.2% in benign, 19.5% in borderline, and 68.3% in malignant group). Statistical analysis revealed a statistically significant positive correlation between p53 immunostaining properties and tumor size (p = 0.01), mitotic index (p=0.05), cellularity (p = 0.042), and diagnosis (p = 0.01). CONCLUSIONS: We conclude that p53 expression may be a useful tool in classifying and predicting the prognosis of gastrointestinal stromal tumors as it is well correlated with tumor size, mitotic index, cellularity, and diagnosis according to our study results. PMID- 17419249 TI - En bloc resection for primary non-Hodgkin's lymphoma of the descending colon with massive extension into the neighboring organs. AB - A 79-year-old female who was surgically treated and received adjuvant chemotherapy for primary non-Hodgkin's lymphoma of the descending colon with massive extension into the pancreatic tail, spleen, and left kidney is herein reported. The patient had acute colonic obstruction and an urgent laparotomy was performed because endoscopic decompression using a transnasal ileus tube could not be done. We performed a left colectomy with an en bloc resection of the neighboring organs, followed by the creation of an end colostomy. The patient had no major postoperative complications and is well without any recurrence of lymphomas 6 months after surgery. PMID- 17419250 TI - Advantages of thoracoabdominal approach by oblique incision for right-side hepatectomy. AB - BACKGROUND/AIMS: Thoracoabdominal approach might be safe and facilitate hepatic resection for tumors located in the right lobe. To evaluate the clinical usefulness of the thoracoabdominal approach using oblique incision for the right side hepatectomy, we compared the perioperative data with those of the abdominal approach. METHODOLOGY: The oblique incision for the thoracoabdominal approach was placed along the intercostal space (Oblique group, n=13). The J-shape incision for abdominal approach consisted of an upper median incision and transverse incision (J-shape group, n=13). RESULTS: Patient demographics were similar in the two groups. Operation time was significantly shorter in the oblique group (292 +/ 122 min) than in the J-shape group (450 +/- 137 min, p < 0.01). The difference was noted regardless of the extent of hepatic resection. Clamping time and blood loss were similar in the two groups. The postoperative period of use of analgesia tended to be shorter in the oblique group (9 +/- 3 days) than in the J-shape group (15 +/- 11 days) but not significant (p = 0.08). Postoperative liver function tests, complications and clinical outcome were not significantly different between the two groups. CONCLUSIONS: Thoracoabdominal approach using oblique incision was useful for resection of liver tumors located in the hepatic dome and posterior segment. PMID- 17419251 TI - Surgical treatment of pulmonary metastasis from hepatocellular carcinoma. AB - BACKGROUND/AIMS: The lung is one of the most common sites of extrahepatic spread from hepatocellular carcinoma (HCC). The aim of this study was to evaluate the efficacy of surgical management in patients with pulmonary metastasis from HCC. METHODOLOGY: Fourteen patients with pulmonary metastases arising from HCC underwent surgery at the National Cancer Center Hospital between 1980 and 2001. The clinical and pathological data were analyzed retrospectively. RESULTS: Four patients were still alive and none of them had evidence of recurrent disease. Ten patients had died with recurrent HCC. The postoperative morbidity and mortality rates were low. The mean overall survival was 42.7 months (range, 8-87). The mean survival after initial thoracotomy was 21.6 months (range, 1-66). The 1-, 2-, and 5-year survival rates after metastasectomy were 71.4%, 44.6%, and 26.8%, respectively. CONCLUSIONS: Surgical treatment is appropriate option in selected patients for pulmonary metastasis arising from HCC, and might prolong survival. PMID- 17419252 TI - Epidemiology of hepatocellular carcinoma in lower Egypt, Mansoura Gastroenterology Center. AB - BACKGROUND/AIMS: Hepatocellular carcinoma (HCC) is now regarded as one of the major malignant diseases worldwide, with significant variations in its epidemiology. OBJECTIVE: study of the epidemiology of hepatocellular carcinoma and its relation to age, sex, residence, and viral hepatitis in Egypt. METHODOLOGY: From January 1992 to May 2005, 1012 cases with hepatocellular carcinoma were diagnosed in the Gastroenterology Center, Mansoura University, Egypt. All patients were evaluated for age, sex, residence, occupation, history of other medical disease, anti-bilharzial treatment, blood transfusion, viral markers, and liver functions. RESULTS: The mean age was 54.26 +/- 9.2, with high prevalence between 51 and 60 years. Male to female ratio was 5:1, farmers constituted 37.6%, workers 22.9% and housewives 12.8% of the patients. The number of HCC patients increases yearly from only 9 patients evaluated at 1992 to 80 patients in the first 5 months of the year 2005. The mean age increased from 45 years at 1992 up to 58 years at year 1996. Seventy-seven percent of the patients were resident in rural areas versus 23% in urban areas. Seventy-eight percent of patients presented with abdominal pain, 7.1% with abdominal swelling, 4.3% with jaundice, 2.4% with other symptoms and 8.8% of the HCCs were discovered accidentally. The prevalence of diabetes mellitus among HCC patients was 13.6%, history of anti-bilharzial treatment was positive in 37.6%. Hepatitis C, B and mixed B&C was 76.6%, 3.3%, 3.6% respectively. History of blood transfusion was encountered in only 10.6%. Twenty-seven percent of HCC patients had previous history of surgery. Clinical and laboratory studies revealed that, 52.1% of patients were Child's A, 37.3% B and 10.7% C. Tumor size >5 cm in 65.5%, the right lobe was the site in 63.9%, left lobe in 25.8% and both lobes in 10.3% of the patients. Lesions were single in 71.1%, multiple in 24.9% and diffuse in 4% of cases. Portal vein thrombosis was detected in 15.9%. Hepatic resection was done in 25.8%, chemoembolizations in 17.2%, radiofrequency in 13.1%, alcohol injection in 0.9%, mixed treatment in 3.5% of patients while 38.7% were managed by conservative treatment due to no available safe treatment. CONCLUSIONS: The number of newly diagnosed patients with HCC increases annually. The prevalence of HCC is high in Nile Delta area, and is more common in males, rural residents and farmers especially in HCV patients. In rural areas there are other risk factors that may be responsible for this high incidence, such as pollution, aflatoxins and use of insecticides, which need more study. PMID- 17419253 TI - Ten-year survival of patients with hepatocellular carcinoma after hepatectomy. AB - BACKGROUND/AIMS: The characteristics in patients with hepatocellular carcinoma who survive more than 10 years after hepatectomy remain unclear. METHODOLOGY: Eighty-five cases of hepatocellular carcinoma were retrospectively identified as short-term survivors (S-group: <5 years, n=41), medium-term survivors (M-group: > or =5 and <10 years, n=25), and long-term survivors (L-group: > or =10 years, n=19) to examine the clinicopathologic factors for the 10-year survival after curative hepatectomy. RESULTS: In the L-group, platelet count and albumin level were higher and total bilirubin level was lower than those in the S-group. In the S-group, the total bilirubin level was higher and vascular invasion was more frequent than those in the M-group. Multiple regression analysis revealed that only host-related factors such as age, albumin level, and total bilirubin level were selected as significant factors to determine the 10-year survival. However, no tumor-related factor was selected. CONCLUSIONS: The present study suggests that the important factor linked to the 10-year survival of hepatocellular carcinoma patients is the host-related factor, but not the tumor-related factor. Especially, younger age, higher albumin level, and lower total bilirubin level are quite important to determine the 10-year survival. PMID- 17419254 TI - Overexpression of MT3-MMP in hepatocellular carcinoma correlates with capsular invasion. AB - BACKGROUND/AIMS: Extracellular matrix-degrading matrix metalloproteinases (MMPs) are invariably up-regulated in epithelial cancers and are key agonists of angiogenesis, invasion and metastasis. Recent studies have shown high levels of various MMPs, including MT1-MMP, MMP-1, MMP-2 and MMP-9, and their involvement in tumor progression in human hepatocellular carcinoma (HCC). However, the expression and role of MT3-MMP in HCC remains unclear. METHODOLOGY: We examined the immunohistochemical expression of MT3-MMP in surgically resected HCCs (n=58), hepatitis C virus (HCV) and hepatitis B virus (HBV)-related chronic hepatitis (n=34) and cirrhosis (n=24). RESULTS: MT3-MMP expression was observed in all non cancerous liver tissues. In HCCs, 52% (30/58) of patients showed high MT3-MMP expression while the remaining 48% (28/58) of patients showed low expression. A clinicopathological survey demonstrated a significant correlation between high MT3-MMP expression and capsular invasion of carcinoma (p = 0.034) although there was no correlation between high MT3-MMP expression in HCC and overall survival or disease-free survival. CONCLUSIONS: MT3-MMP was expressed not only in chronic hepatitis and liver cirrhosis, but also in HCC, and high MT3-MMP expression correlated significantly with capsular invasion of carcinoma. PMID- 17419255 TI - Hepatic resection followed by IFN-alpha and 5-FU for advanced hepatocellular carcinoma with tumor thrombus in the major portal branch. AB - BACKGROUND/AIMS: The prognosis of hepatocellular carcinoma (HCC) invading the major branches of the portal vein (Vp3) is extremely poor. Recently, we reported the efficacy of combination therapy with subcutaneous interferon (IFN)-alpha and intra-arterial 5-FU for intractable HCC with Vp3. In this study, this therapy was applied for resectable advanced HCC (Vp3) as a postoperative adjuvant. METHODOLOGY: Patients with HCC and tumor thrombi either in the major or first branch of portal vein were included (n=30). Fifteen consecutive patients with HCC and Vp3 were treated with at least 3 cycles of a combination therapy consisting of continuous arterial infusion of 5-FU (300 mg/mm3/day, 5 days/week, for the initial 2 weeks) and subcutaneous injection of IFN (5 MIU, 3 times/week, 4 weeks) as a postoperative adjuvant therapy following hepatic resection. Another 15 patients who underwent hepatic resection with no IFN/5-FU chemotherapy acted as controls. RESULTS: The results were as follows in the IFN/5-FU adjuvant treatment group; disease-free survival (n=11, 5-55 months), survival with recurrence (n=2, 9, 48 months), cancer death (n=1, 18 months), death from other causes but no recurrence (n=l, 22 months). The 1-year survival rate was 100% in patients treated with IFN/5-FU, and 41% in those without IFN/5-FU historical controls (n=15). There was a significant difference in disease-free and overall survival rates between the two groups (p = 0.0033 and 0.0031). CONCLUSIONS: Combination therapy with subcutaneous IFN and intra-arterial perfusion of 5-FU seems to be a promising postoperative adjuvant treatment modality for resectable HCC with Vp3. PMID- 17419256 TI - Quantitative analysis of Doppler waveform of hepatic veins for assessment of hepatic functional reserve. AB - BACKGROUND/AIMS: The Doppler waveform of hepatic veins in healthy humans is a triphasic waveform (two negative waves and one positive). Recent studies emphasize the role of Doppler ultrasonography in the evaluation of liver cirrhosis. The analysis of the Doppler waveform of hepatic veins might be a useful and non-invasive method to assess hepatic functional reserve and the amount of the liver to be excised. METHODOLOGY: In 30 patients scheduled for hepatectomy, we performed quantitative analysis of the Doppler waveform of hepatic veins and correlated the Pulsatility Index (PI) and Resistance Index (RI) with ICG clearance values and histologic findings of resected liver specimens in order to ascertain the clinical utility of this method. RESULTS: The mean (+/-SE) values of PI and RI were 1.31 +/- 0.10 and 0.82 +/- 0.03, respectively, and both values are diminished according to the severity of cirrhosis and very closely correlated with fibrosis of the liver-reflected hepatic consistency (PI: r = 0.611, p = 0.0010; RI: r = -0.647, p = 0.0005). They were also correlated with ICG clearance values to some extent, but several discrepant cases were observed. In four of the discrepant cases (ICGR15 < or = 10%, but PI and RI or =10 cm), cyst infection, multivesicular content, solitary cysts and location in the left lobe of liver as well as long duration of symptoms as independent predictors of frank IBR. CONCLUSIONS: Patients with large (> or =10 cm), multivesicular, solitary, left lobar or infected HHC as well as those with prolonged history are at increased risk of frank IBR and should be offered early surgery. PMID- 17419264 TI - Treatment of advanced hepatocellular carcinoma with gemcitabine plus oxaliplatin. AB - BACKGROUND/AIMS: To investigate the effects of gemcitabine-oxaliplatin in patients with advanced unresectable hepatocellular carcinoma (HCC). METHODOLOGY: Forty patients confirmed with unresectable, nonembolizable HCC and objective measurable tumors, had received no prior systemic chemotherapy. All patients enrolled in this study had adequate liver and renal functions, and adequate bone marrow reserve. Gemcitabine (1250 mg/m2) was given on day 1 and oxaliplatin (100 mg/m2) on day 2 via intravenous infusion in each 21-day cycle. If no evidence of disease progression or unacceptable adverse effects occurred, the treatment courses would continue. RESULTS: No affirmative response was achieved; partial response (PR) was achieved in 1 patient. Eight patients (20%) had stable diseases with an average duration of 20.2 weeks. The others were rated progression. The average time to progression was 13.9 weeks. The spectra of both hematologic and nonhematologic toxicities were mild, with thrombocytopenia as the dose-related side effect. CONCLUSIONS: The toxicities of gemcitabine-oxaliplatin were well managed in this study. In view of these treatment results, gemcitabine oxaliplatin combination therapy with this particular dose regimen should not be considered in patients with advanced hepatocellular carcinoma. PMID- 17419265 TI - Fertile females with nonalcoholic fatty liver disease (NAFLD) have higher levels of ALT than postmenopausal females: implications for the influence of fertility on NAFLD. AB - BACKGROUND/AIMS: Insulin resistance recently has been reported to play a major role in nonalcoholic fatty liver disease (NAFLD). We evaluated the influence of fertility on fatty liver injury in fertile and postmenopausal women with insulin resistance. METHODOLOGY: We investigated 152 patients with noninsulin-dependent diabetes mellitus without insulin treatment; 46 males, 52 fertile women and 54 postmenopausal women. All had liver damage and/or steatosis recognized by ultrasonography. We measured the fasting serum levels of C-peptide and insulin, as markers of insulin resistance, and the serum levels of ALT. The severity of liver steatosis was judged by ultrasonography. RESULTS: Fertile females had significantly higher levels of ALT and demonstrated a more significant correlation between serum levels of ALT and C-peptide or insulin than did the postmenopausal females or males. Fertile females with moderate to severe steatosis had significantly higher levels of ALT than those with mild or no steatosis, although such a significant difference was not found in postmenopausal females or males. CONCLUSIONS: We demonstrate that fertility is an important factor in fatty liver damage of NAFLD with insulin resistance, suggesting that estrogen may exacerbate nonalcoholic steatohepatitis. PMID- 17419266 TI - The effects of somatostatin and ursodeoxycholic acid in preventing the ischemic injury of the liver following Pringle maneuver in obstructive jaundice-rat model. AB - BACKGROUND/AIMS: In our study, the effects of somatostatin (SS) and ursodeoxycholic acid (UDCA) on ischemic liver injury were studied in (obstructive) jaundice-rat model. METHODOLOGY: For this purpose, jaundice was produced in the first four groups by binding of their choleducts. We performed just laparotomy to the other four groups of animals. To groups 1 and 5, SS was given 15 mcg/kg/day intraperitoneally, and to groups 2 and 6, UDCA was given 20 mg/kg/day enterally. No drugs were given to any other group. At the end of one week, a procedure with ischemia of the liver for 60 minutes followed by reperfusion for 2 hours, was performed to each rat except for groups 4 and 8. Following this procedure, they were sacrificed. The blood samples were taken to measure SGOT, SGPT, ALP, LDH, total and direct bilirubin levels, while liver biopsies were taken for histopathological evaluation. RESULTS: Under normothermic conditions, following 60-minute liver ischemia period, no irreversible histopathological changes were detected. However, increases in liver necrosis parameters were noted biochemically. SS and UDCA were thought to be effective in preventing the injury by decreasing the liver enzymes levels to a significant degree. The damage of the hepatic ischemic injury was found to be more meaningful and prominent in liver with jaundice. CONCLUSIONS: In this study, it was noted that SS and UDCA decrease the effects of cholestatic hepatic injury especially and improve the condition. PMID- 17419267 TI - The feasibility of Makuuchi criterion for resection of hepatocellular carcinoma. AB - BACKGROUND/AIMS: The Makuuchi criterion was proposed to select for the appropriate hepatectomy in an impaired liver. However, there are no comparative analyses concerning this criterion. Our purpose is to evaluate the efficacy of it. METHODOLOGY: We conducted our study on 471 patients with hepatocellular carcinoma, resected from January 1986 to April 2004. We defined surgery consistent with Makuuchi criterion as group 1, and other as group 2. We performed comparative analysis of surgical results. RESULTS: Group 1 showed lower intraoperative blood loss (1602 +/- 119 mL vs. 2448 +/- 244mL: P = 0.002), mortality (4.4% vs. 12.5%: P = 0.009), and postoperative serum total bilirubin (3.4 +/- 0.2 mg/dL vs. 6.2 +/- 0.8 mg/ dL: P < 0.0001) than group 2. Five-year survival rates were not significantly different between group 1 (43%) and 2 (35%): P = 0.18. Blood loss in operative deaths was greater than in survivors, both in group 1 (6155 +/- 1725 mL vs. 1391 +/- 83 mL: P < 0.0001) and in group 2 (4381 +/- 946 mL vs. 2172 +/- 231mL: P = 0.002). Blood loss in patients with postoperative liver failure was also greater than in others, both in group 1 (2692 +/- 292 mL vs. 1146 +/- 106 mL: P < 0.0002) and in group 2 (2968 +/- 335 mL vs. 1538 +/- 265 mL: P = 0.004). CONCLUSIONS: Our study showed Makuuchi criterion is helpful for a safe hepatectomy for hepatocellular carcinoma. If inconsistent with it, 2172 mL and 1538 mL blood loss are considered permissible upper limits to avoid operative death and liver failure, respectively. PMID- 17419268 TI - Triple cancer in liver, esophagus and urinary bladder: indications and problems of hepatectomy. AB - In patients with multiple cancers and chronic liver damage, it is difficult to decide on the operative procedures. We describe a 70-year-old man with triple cancer: liver, esophageal and urinary bladder. The patient had the complication of chronic hepatitis C and mild liver damage. He received curative resections of all cancers. In patients with multiple cancers, including hepatocellular carcinoma accompanied by liver damage, the treatment strategy should be decided after clarifying the relationship between the staging of multiple cancers and liver function. PMID- 17419269 TI - Thoracoscopy-assisted radiofrequency ablation liver cancer. Report of three cases. AB - To achieve complete ablation of liver cancer in the hepatic dome, thoracoscopy assisted radiofrequency ablation was attempted in three patients. Under general anesthesia and left lateral position, a thoracoport site was placed in the 6th intercostal space and a small thoracotomy was placed in the 7th intercostal space. In all three cases, the liver cancer was located in the hepatic dome of segment 7/8 and the tumor could not be fully observed on the lung echogram. In Case 1, saline was infused into the thoracic cavity under thoracoscopic observation and percutaneous ablation was safely performed. In Cases 2 and 3, the right diaphragm was opened, guided by ultrasonography, and the electrode was inserted into this working space via the thoracotomy site. At day 7, complete ablation was confirmed by computed tomography. All patients recovered and were discharged after a short hospital stay without severe complications. Tumor recurrence has not been observed in any patient at this stage. For liver cancer located in the right subphrenic dome of the liver, thoracoscopy-assisted ablation is a safe and useful option particularly in patients with poor hepatic function. PMID- 17419270 TI - Long-term follow-up study of surgical treatment for pancreatic stones. AB - BACKGROUND/AIMS: With rare incidence and lack of extensive study for pancreatic stones, some issues in this area remain to be clarified. Surgical experience for pancreatic stones was presented, particularly focusing on the surgical risk and long-term outcome. We also evaluated the role of the pancreatic stone in pancreatitis and pancreatic cancer. METHODOLOGY: Data of patients with pancreatic stones are analyzed between 1984 and 2002, with a median follow-up period of 67 months. Clinical features and characteristics of pancreas and pancreatic stone are evaluated. Diagnostic image studies are compared. Outcome measures are surgical risks including surgical morbidity and mortality, and degree of long term symptom control. RESULTS: There were 18 patients with pancreatic stones. The etiology was idiopathic in 50% of cases, and alcoholism in 33.3%. Abdominal pain was the most common (100%) clinical presentation. Pancreatic cancer was found in 4 (22.2%) patients. Most (61.1%) of the pancreatic stones were located in the pancreatic head. Only 1 patient had a single pancreatic stone, and 12 (66.7%) patients had more than 3 pancreatic stones. All the patients except one (94.4%) presented pictures of chronic pancreatitis. Surgical complication occurred in 2 (11.8%) patients, and surgical mortality in 1 (5.9%) resulting from pneumonia. Improvement of clinical symptoms after surgery was achieved in nearly all (93.8%) patients, including 56.3% free of symptoms, 25.0% much improvement and 12.5% mild improvement. CONCLUSIONS: Removal of pancreatic stones combined with surgical drainage of pancreatic duct or resection of pancreas might have symptomatic benefits. Surgical intervention is recommended for all patients with pancreatic stones, in terms of symptom relief, cancer risk and low surgical risk. PMID- 17419271 TI - The role of octreotide versus placebo in the prevention of post-ERCP pancreatitis. AB - BACKGROUND/AIMS: To evaluate the effectiveness of a single administration of intravenous octreotide infusion in preventing post-ERCP pancreatitis and progressing hyperamylasemia. METHODOLOGY: One hundred and twenty (71 female, 59 male) patients who had been diagnosed with pancreaticobiliary pathology were included in this study. 100 microgram (0.1 mg) octreotide diluted in 60 mL normal saline solution administered intravenously 60 minutes prior to the procedure and continued during the procedure and after the procedure. Placebo was given in 87 patients. Patients were assessed clinically and serum amylase level was also measured before the procedure and 3, 12, and 24 hours after the procedure. We define clinical pancreatitis as serum amylase level greater than 4-5 times in conjunction with clinical assessment. RESULTS: Hyperamylasemia was assessed in 14 of 33 (42.4%) administered octreotide patients. Clinical findings of pancreatitis were observed in 5 of these 14 (11.5%) patients. Hyperamylasemia was also assessed in 41 of 87 (47.1%) administered placebo patients. Clinical findings of pancreatitis were observed in 10 of these 41 (11.5) patients. There were no significant differences between the groups, statistically (p > 0.05) (Pearson chi square test). CONCLUSIONS: The results of this trial indicate that a single administration of intravenous octreotide infusion does not prevent ERCP-induced pancreatitis and effect serum amylase level. PMID- 17419272 TI - Predictors of systemic chemotherapy contraindication in pancreatic cancer patients with distant metastasis. AB - BACKGROUND/AIMS: To investigate predictors of systemic chemotherapy contraindication in pancreatic cancer patients with distant metastasis. METHODOLOGY: Eighty-seven consecutive pancreatic cancer patients with distant metastasis receiving systemic chemotherapy using 5-fluorouracil, cisplatin or gemcitabine were analyzed retrospectively to investigate prognostic factors. RESULTS: The overall median survival time of all patients in the whole series was 3.8 months, the 3-, 6, and 12-month probability of survival being 58%, 26%, and 3%, respectively. Significant poor prognostic factors were the age of 65 years old or older, presence of ascites, a total bilirubin > 2.5 mg/dL, ChE <110 IU/L, a higher level of tumor maker (CA19-9) and performance status <80 (p < 0.005). Cox proportional hazards model revealed independent poor prognostic factors were a presence of ascites, serum ChE level <110 IU/L, and age > or =65. A prognostic index was calculated based on the regression coefficients derived from the three variables according to their relative risk of death (RRD) = exp (presence of ascites x 1.213 + serum ChE level x 1.065 + age > or = 65 x 0.651). CONCLUSIONS: Ascites, ChE <110 IU/L, or age > or =65 should be chemotherapy contraindications for pancreatic cancer patients with distant metastasis because of their extremely short survival time and some other experimental approaches or supportive care are needed. PMID- 17419273 TI - EUS FNA in intraductal papillary mucinous tumors of the pancreas. AB - BACKGROUND/AIMS: There is little information concerning the potential role of fine-needle aspiration guided by endoscopic ultrasonography in the pathologic diagnosis of intraductal papillary mucinous tumors of the pancreas. METHODOLOGY: Patients with an intraductal papillary mucinous tumor of the pancreas suggested by endoscopic ultrasonography underwent fine-needle aspiration guided by endoscopic ultrasonography in order to investigate the presence of mucin and/or cytologic changes consistent with this diagnosis. A group of 111 patients with other pancreatic lesions explored during the same period of time was used as a control group. RESULTS: Fine-needle aspiration guided by endoscopic ultrasonography was safely performed in 19 patients and supported the diagnosis in 17 of them. Nine out of the 17 patients with suspicion of intraductal papillary mucinous tumors of the pancreas went to surgery and this diagnosis was confirmed in the resected specimen in all of them. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy of EUS FNA in the diagnosis of IPMT were 82%, 100%, 100%, 92% and 94% respectively. CONCLUSIONS: Fine-needle aspiration guided by endoscopic ultrasonography is a good technique to support the diagnosis of intraductal papillary mucinous tumors of the pancreas and should be considered in this group of patients if pathologic confirmation is judged to be necessary. PMID- 17419274 TI - Laparoscopic distal pancreatectomy: a comparative study. AB - BACKGROUND/AIMS: This article aims to describe the different techniques of laparoscopic distal pancreatectomy and to compare the results of our series of 9 laparoscopic resections against the historical open control in the same institution. With the advent of laparoscopic surgery, there is an increasing number of patients with different pancreatic pathologies that can now be managed by minimal access surgery. The initial results of laparoscopic pancreatectomy are quite promising particularly for those small neuroendocrine and cystic neoplasms located at the body and tail of pancreas. METHODOLOGY: The different techniques of laparoscopic distal pancreatectomy are described in detail with special emphasis on the need of "hand assistance" and the different methods of splenic preservation. The perioperative data of 9 laparoscopic distal pancreatectomies are analyzed and compared against the 5 historical open controls in the same institution. RESULTS: There were 9 laparoscopic pancreatic resections performed in our institution since 1999. Indications for surgery included 5 cystic neoplasms (1 patient with concomitant splenic artery aneurysm), 1 chronic pancreatitis with pancreatic duct stricture and a small pseudocyst, 1 pseudopancreatic tumor secondary to seal off perforated posterior gastric ulcer, 1 pseudopapillary tumor and 1 neuroendocrine tumor. There were 6 females and 3 males with median age of 61 years (range 18-79). The majority of patients was of low anesthetic risk (ASA 1 or 2). Total laparoscopic resection was performed in 7 cases and 2 resections were performed using the hand-assisting technique. Out of the 4 cases with splenic preservation, only one patient had both splenic artery and vein successfully preserved, whereas the other 3 cases had to rely on the short gastric arcade. Median operating time was 180 minutes (range 120-250) and median blood loss was 100cc (range 50-500). Pancreatic leak occurred in two patients (22.2%) and 1 patient developed intraabdominal collection, all of which settled upon conservative treatment. In our series, clear resection margin was obtained for all the neoplastic cases. Median hospital stay was 7 days (4-53). Postoperatively, patients consumed an average of 15 tablets of dologesic. No other complications were observed upon a median follow-up of 15 months (1-50). When results were compared to the 5 historical open controls (excluding those malignant tumors), patients managed with this new approach had significantly less intraoperative blood loss (100 vs. 450 mL, P = 0.021). CONCLUSIONS: Our initial experience not only confirmed the feasibility oflaparoscopic pancreatectomy, but also demonstrated the promising results of this approach in selected patients. PMID- 17419275 TI - Analysis of clinicopathological features and predictors of malignancy in intraductal papillary mucinous neoplasms of the pancreas. AB - BACKGROUND/AIMS: Intraductal papillary mucinous neoplasms (IPMNs) are increasingly recognized, but it is very difficult to evaluate accurately the malignancy of these neoplasms by modern imaging. We reviewed our experience in order to elucidate predictors of tumor malignancy, invasiveness, and outcome. METHODOLOGY: The clinicopathological features and surgical outcomes of 57 patients with IPMNs who underwent surgery in Nagoya University Hospital were analyzed. RESULTS: The histological diagnosis was adenoma in 40, borderline in 1, carcinoma in situ (CIS) in 7, and invasive carcinoma in 9 patients. Patients with invasive carcinomas had significantly shorter survival rates than patients with benign IPMNs or CIS (p < 0.0001). Multivariate analyses revealed that the main duct or the combined type was significantly predictive of malignancy, and both main duct or combined type and diabetes mellitus were associated significantly with invasive carcinoma. CONCLUSIONS: IPMNs generally grow slowly, but have a malignant potential that warrants radical surgical treatment when the tumor component invades the parenchyma. Our results suggest that the above factors should be considered in surgical management. The main duct type of IPMN or IPMN with mural nodules is potentially malignant or invasive. Therefore, radical operative management is indicated in these IPMNs. PMID- 17419276 TI - Current medical treatment of pancreatic neuroendocrine tumors. AB - Pancreatic neuroendocrine tumors (NETs) consist of a wide group of neoplasms, with different biological behaviors in terms of aggressiveness and hormone production. In the last two decades, significant progress has been observed in our understanding of their biology, diagnosis and treatment. Surgery remains to be the only curative approach, but unfortunately the diagnosis is often delayed due to the slow growth of these tumors and the difficulty in identifying the symptoms related to the tumor-released hormones. In addition to surgery, other approaches to control the disease are biological therapy consisting of somatostatin analogs and interferon (IFN), systemic chemotherapy, radioligand therapy and local therapy with chemoembolization. Several newer cytotoxic agents, including irinotecan, gemcitabine, taxanes, oxaliplatin, capecitabine and PS-341 have been studied in metastatic patients. Considering the high vascularity of these tumors, antiangiogenic agents like endostatin and thalidomide have also been evaluated in advanced NETs. Although these agents seem to have potential activity in NETs and may increase progression free survival, none of these currently available medical therapeutic options are curative. While more efficient novel strategies are to be developed, the rationale use of the current therapeutic options may improve quality of life, control the symptoms related to the hypersecretion of hormones and/or peptides, control tumor proliferation and prolong survival in patients suffering from NETs. PMID- 17419277 TI - Distribution of gastric lymphoid follicles in Helicobacter pylori-associated gastritis. AB - BACKGROUND/AIMS: Lymphoid follicles and the satellite lesions (intestinal metaplasia, atrophy and dysplasia) are known as precursor lesions of mucosa associated lymphoid tissue lymphomas in gastritis. Little is known about their prevalence in different distributions and types of Helicobacter pylori-associated gastritis. The aim of the study was to estimate the topographic prevalence of these lesions in gastritis related with Helicobacter pylori and to associate them with the density of bacteria. METHODOLOGY: Histology for the type of gastritis and for lymphoid follicles and Helicobacter pylori density were studied in antrum and/or corpus biopsies taken from 107 consecutive patients with clinical diagnosis of peptic ulcer. RESULTS: Lymphoid follicles, panmucosal and superficial gastritis were seen in 31 (31.9%), 84 (86.6%) and 13 (13.4%) out of 97 antrum biopsies, respectively. In the corresponding 28 corpus biopsies, these lesions were seen in 8 (28%), 15 (54%), 13 (46%), respectively. Lymphoid follicles were found more in panmucosal than superficial gastritis in the antrum, however in the same ratios in the corpus. In association with lymphoid follicles, Helicobacterpylori was positive in 7 (87%) of 8 corpus biopsies and in all (100%) of 31 antrum biopsies. No relation was observed between lymphoid follicles and Helicobacter pylori density. CONCLUSIONS: Examination of antrum biopsies rather than corpus biopsies would be sufficient to screen precarcinogenic lymphoid follicles in Helicobacterpylori associated gastritis. PMID- 17419278 TI - 5-fluorouracil, mitomycin-C, and polysaccharide-K adjuvant chemoimmunotherapy for locally advanced gastric cancer: the prognostic significance of frequent perineural invasion. AB - BACKGROUND/AIMS: Although adjuvant chemotherapy has demonstrated small but significant survival benefit in locally advanced gastric cancer in several meta analyses, optimal chemotherapy regimen remains to be determined. METHODOLOGY: We retrospectively analyzed the survival of 207 gastric cancer patients (stage IB: 19, II: 65, IIIA: 58, IIIB: 28, IV: 37) who underwent 5-fluorouracil (5-FU), mitomycin-C (MMC), and polysaccharide-K (PSK) chemoimmunotherapy (CITX) after curative resection (FM group). The survival of FM group was compared with that of historical control cohort of 103 patients with almost identical stage distribution who received 5-FU and doxorubicin-based chemotherapy (FA group). RESULTS: Five-year disease-free survival and overall survival (OS) of FM group were 58.7% and 59.1%, respectively. Frequent perineural invasion was significantly associated with poor OS (p = 0.01) in multivariate analysis. There was no statistically significant difference in 5-year OS (59.1% vs. 56.2%, p = 0.637) between FM and FA groups. FM group showed superior 5-year OS (84.4% vs. 67.6%, p = 0.019) compared with FA group in stage IB or II patients without significant difference (p = 0.222) in stage IIIA to IV. CONCLUSIONS: 5-FU, MMC, and PSK CITX is as effective as 5-FU and doxorubicin-based chemotherapy. Moreover, frequent perineural invasion seems to be an important poor prognostic factor. PMID- 17419279 TI - Palliative surgery for advanced stage (stage IV) gastric adenocarcinoma. AB - BACKGROUND/AIMS: The factors that effect resectability, major morbidity, early mortality, and survival in advanced stage gastric adenocarcinoma patients are evaluated. METHODOLOGY: Records of 74 patients that underwent surgery for stage IV gastric adenocarcinoma in the Third Surgical Clinic of Izmir Ataturk Training Hospital between January 1997 and January 2004 were reviewed retrospectively. Two groups (the patients with resectable disease and those with unresectable disease) were compared with regard to age, gender, primary complaint, symptoms, site of the tumor, involvement of adjacent organs, lymph node involvement, distant metastases, differentiation of tumor, surgical procedure, perioperative blood transfusions, and postoperative hospital stay. RESULTS: Mean age of the 74 patients was 58.4 years. Forty-five cases (60.8%) were considered as unresectable and 29 (39.2%) patients underwent a palliative resection. There was a significant relation between resectability and site of the tumor, and severity of invasion. As the number of perioperative blood transfusions increased, morbidity and mortality increased significantly. Although early mortality was high in the palliative resection group, survival (mean 10.4 months, longest 25 months) was better compared to that of the unresectable gastric cancer group (mean 3.5 months, longest eight months). CONCLUSIONS: A palliative gastric surgery may be applied to improve prognosis of advanced gastric cancer patients, even at the presence of peritoneal dissemination, hepatic metastases, N3 lymph node involvement, adjacent organ invasion, or poor differentiation of the tumor. PMID- 17419280 TI - Results of follow-up endoscopy in patients who underwent proximal gastrectomy with jejunal interposition for gastric cancer. AB - BACKGROUND/AIMS: The present study evaluates the findings of long-term follow-up endoscopy in patients who underwent proximal gastrectomy with jejunal interposition for gastric cancer. METHODOLOGY: A total of 55 patients (45 males, 10 females; 32 to 79 years; mean, 55.9 years), who underwent proximal gastrectomy with jejunal interposition, were enrolled in the present study. We reviewed the findings of follow-up endoscopy of all patients with particular reference to the development of esophagitis, jejunitis, jejunal ulcer and secondary tumors. RESULTS: We found reflux esophagitis in 6 patients (10.9%) between 12 and 35 months with an average of 22 months after surgery. Jejunitis was discovered in 5 patients (9.0%) between 6 and 96 months with an average of 29 months after surgery. Jejunal ulcer was revealed in 6 patients (10.9%) between 6 and 75 months with an average of 37 months after surgery. Tumors of the remnant stomach, early gastric cancer and gastric adenoma, were identified in 2 patients (3.6%) at 24 months and 69 months, respectively. CONCLUSIONS: Jejunal interposition combined with proximal gastrectomy does not always prevent complications related to regurgitation of gastric content, and may not be a suitable treatment in view of postoperative endoscopic surveillance. Further studies are required to identify an appropriate surgical approach to proximal gastrectomy for gastric cancer. PMID- 17419281 TI - Clinicopathological and immunohistochemical markers after radical gastrectomy for gastric cancer. AB - BACKGROUND/AIMS: The aim of the study was to verify the classic clinicopathological prognostic factors of the expected survival, to determine the frequency of immunohistochemical factors (C-erB-2, p53) and to verify these immunohistochemical factors in their function as prognostic factors in gastric cancer patients after a potentially curative gastric resection. METHODOLOGY: In 78 gastric cancer patients after a potentially curative resection the clinicopathological factors (age, general performance of the patient, tumor diameter, TNM stage, stage according to UICC, type of operation and the extent of lymphadenectomy) and immunohistochemical factors (p53 and C-erB-2) were analyzed. RESULTS: In univariate analysis ASA stage of general performance, size of tumor, TNM stage and stage according to UICC exert significant influence on the median survival. Median survival is higher in patients with C-erB-2 negative reaction and in those with p53 positive reaction, although not significant. In patients with tumor located in proximal two thirds, patients with negative reaction to C erB-2 have a significantly higher median survival than patients with positive reaction to C-erB-2. The Cox regression model reveals ASA stage and UICC classification to be significant factors whereas immunohistochemical parameters C erB-2 and p53 do not reveal any significance in the multivariate survival analysis. CONCLUSIONS: According to the results of our study the ASA stage and UICC classification influence the expected survival of potentially radically resected gastric cancer patients significantly, while immunohistochemical parameters C-erB-2 and p53 do not. PMID- 17419282 TI - Expression of MMP-9 and TIMP-1 as prognostic markers in gastric carcinoma. AB - BACKGROUND/AIMS: The aim of the present work is to clarify the role of metalloproteinase-9 and its inhibitor in the evolution of gastric cancer after surgical resection. METHODOLOGY: We have studied 44 gastric cancer patients submitted to surgery. There were 13 proximal tumors, 16 located in the middle third and 15 in the distal one. Overall survival was 26% at 6 years. Metalloproteinase-9 and tissue inhibitor of metalloproteinase concentrations were investigated by means of ELISA in frozen samples of tumoral and normal gastric mucosa. RESULTS: Mean concentration of metalloproteinase-9 in tumoral tissue was 42 ng/mg of total protein, and this value was 6.9 times greater than the mean concentration in non-tumoral tissue. Cancer tissue also expressed higher levels of TIMP-1, 7.25 versus 4.39 ng/mg of protein. Higher levels of metalloproteinase expression in tumoral tissue, greater [metalloproteinase in tumor]/[metalloproteinase in non-tumor] ratio and greater [metalloproteinase]/[inhibitor] ratio in tumor cells, were all of them statistically related to a worse prognosis when T1 and T2 tumors were considered. CONCLUSIONS: The expression of metalloproteinase-9 or its inhibitor is related to a more aggressive phenotype of gastric cancer. PMID- 17419283 TI - Adjuvant effect of vitamin C on omeprazole-amoxicillin-clarithromycin triple therapy for Helicobacter pylori eradication. AB - BACKGROUND/AIMS: To test the impact of vitamin C supplementation on triple therapy for H. pylori eradication. METHODOLOGY: A total of 171 H. pylori-infected patients were randomized to receive different one-week triple therapies, including 20 mg omeprazole, 1 g amoxicillin, plus the following twice daily: (1) 250 mg clarithromycin (C250 group, n=55); (2) 250 mg clarithromycin and 500 mg vitamin C (V-C250 group, n=61); (3) 500 mg clarithromycin (C500 group, n=55). Six weeks after treatment, the success of H. pylori eradication was assessed by a 13C urea breath test. Each collected H. pylori strain was defined as either clarithromycin susceptible or resistant by E-test. RESULTS: The demographic background, clarithromycin susceptibility of H. pylori, and drug compliance were similar among the three groups (p=NS). For clarithromycin susceptible infection, the V-C250 group had a higher eradication rate than the C250 group (ITT: 85% vs. 68% and PP: 90% vs. 73%, p = 0.03), but had an equivalent rate to the C500 group (p=NS). For clarithromycin resistant infection, all three groups had a similarly poor eradication rate of less than 34%. CONCLUSIONS: Adding vitamin C to one-week triple therapy can reduce the dosage of clarithromycin, but preserve the high eradication efficacy for clarithromycin susceptible H. pylori infection. PMID- 17419284 TI - Endoscopic ultrasonography with three miniature probes of different frequency is an accurate diagnostic tool for endoscopic submucosal dissection. AB - BACKGROUND/AIMS: Endoscopic ultrasound (EUS) examinations are necessary to determine the vertical cancer invasion depth. In this study, we prospectively performed EUS with three miniature probes of different frequency (12, 20, and 30 MHz) in patients with early gastric cancer scheduled for endoscopic submucosal dissection (ESD) based on endoscopic findings. METHODOLOGY: The study included 142 early gastric cancer lesions in 132 patients in our hospital. We then evaluated the diagnostic accuracy of the use of three miniature probes of different frequency in the determination of vertical margins. Finally, we evaluated the value of EUS as an additional examination tool used along with conventional endoscopic findings for determination of indication for ESD. RESULTS: The accuracies of the early gastric cancer invasion depths of the m+sm1 group were 81.0%, 86.0% and 92.3% as measured by the 12-, 20- and 30-MHz probes, respectively. Even though ulceration was associated with the lesions, the 30-MHz probe was the most accurate as compared to the 12- or 20-MHz probes in the m+sml cancer group. CONCLUSIONS: EUS examinations with probes of varying frequency are useful for determining appropriate treatment strategies for early gastric cancer. PMID- 17419285 TI - Genetic characterisation of the cjaAB operon of Campylobacter coli. AB - We investigated the regulation of the cjaA and cjaB genes of Campylobacter coli. These genes are seemingly arranged into one operon but appear to encode functionally different proteins i.e. an extracytoplasmic solute receptor and a MHS - metabolite: H+ symporter transport protein. Analysis of various transcriptional cjaA and/or cjaB lacZ fusion constructs revealed that both genes are arranged in an operon. RACE analysis located the transcription start site of the cjaAB operon 46 bp upstream of the translation start point. Beta galactosidase reporter assays yielded much higher activity for the cjaA than the cjaB gene fusion products. RT-PCR showed unequal amounts of mRNA, indicating differential post-transcriptional processing of cjaA and cjaB mRNA possibly related to the presence of inverted repeats in the intergenic region. Phylogenetic analysis grouped CjaB into a new MHS sub-family together with potential transporters with uncharacterised functions of Campylobacter and Helicobacter. Notably, no CjaB family members were identified in epsilon Proteobacteria from different ecological niches, such as H. hepaticus and Wolinella succinogenes. PMID- 17419286 TI - Natural mannose-binding lectin (MBL) down-regulates phagocytosis of Helicobacter pylori. AB - Considering the role of lectin-carbohydrate interactions between Helicobacter pylori bacteria and the host cells we addressed the question on how mannose binding lectin - MBL, present in human plasma, may influence the phagocytosis of H. pylori by peripheral blood granulocytes. For phagocytosis assay the granulocytes separated from peripheral blood of healthy H. pylori-seronegative donors were used. Phagocytosis was estimated by fluorescence assay using FITC labelled H. pylori cells. The MBL level in the serum samples as well as MBL binding to H. pylori bacteria were estimated by ELISA. In this study all H. pylori isolates bound recombinant mannose binding lectin-MBL as shown by ELISA. The ingestion of H. pylori bacteria in the medium with human serum depleted in natural MBL (nMBL) was more intensive than in the medium with complete serum containing nMBL. Moreover, the ingestion of H. pylori bacteria in the medium with complete serum was increased by an addition of anti-rMBL IgG. The results indicate that interaction of bacterial and host lectins may regulate the phagocytosis of H. pylori bacteria and in this way influence an outcome of the infection caused by these microbes. PMID- 17419287 TI - Phenotypic and genotypic characteristics of Pseudomonas aeruginosa strains isolated from hospitals in the north-west region of Poland. AB - A total of 90 Pseudomonas aeruginosa strains isolated from 4 hospitals in the west-north region of Poland were studied by arbitrarily primed polymerase chain reaction (AP-PCR). AP-PCR results revealed the presence of 11 main groups of patterns (A-K) and 5 unique patterns among isolates. Generally, they were characterized by high resistance to antibiotics tested and significant differences in serogroups and types of growth on Cetrimide Agar medium. It was observed that clonally related strains were isolated from patients within the same ward, among different wards as well as in distant hospitals. PMID- 17419288 TI - Detection of enterotoxic Bacillus cereus producing hemolytic and non hemolytic enterotoxins by PCR test. AB - Nine strains belonging to Bacillus cereus group has been isolated from food and environmental samples. Their taxonomic position was confirmed by RFLP analysis of 16S rRNA gene digested with TaqI. The detection of DNA sequences encoding the hemolysin BL complex and enterotoxin NHE, was studied in Bacillus sp. isolates. Set of primers was used to amplify fragment of hblD gene by PCR. For the detection of nheB gene a new primer set was developed which allowed to amplify 273 bp fragment from wide number of strains belonging to B. cereus group. The hblD gene was present in 7 out of 9 isolates whereas nheB gene occurred in all of them. Reference strains of B. cereus LOCK 0807, and B. thuringiensis NCAIM 01262 contained both genes. Strains of B. subtilis ATCC 6633 and B. pumilus LOCK 0814 do not contain both genes. Obtained results showed that B. thuringiensis NCAIM 01262 contains both genes and therefore may be harmful for human beings. PMID- 17419289 TI - Optimization and purification of alkaline proteases produced by marine Bacillus sp. MIG newly isolated from eastern harbour of Alexandria. AB - A marine Bacillus strain was isolated from the eastern harbour of Alexandria and identified as Bacillus sp. MIG. Maximum activity of studied proteases was obtained when the bacterium was grown in medium with 1% wheat bran and 0.5% yeast extract in addition to the mineral salts and incubated for 48 h at 30 degrees C and 120 rpm. Two alkaline proteases (Pro 1 and Pro 2) were purified to homogeneity using cation exchange chromatography on CM-Sepharose CL-6B followed by Sephadex G-75 superfine. The optimum activities were at pH 11 or 12, and temperatures of 50 and 55 degrees C for Pro 1 and Pro 2 respectively. These two enzymes were relatively stable over pH range from 7.0- 11. Pro 2 was found to be more stable at 50 degrees C in absence of Ca2+ and retained about 47% of its activity after 3 h at this temperature, while Pro 1 lost its activity completely at the same conditions. The two enzymes were active against haemoglobin and casein; in addition, Pro 2 exhibited moderate activity against keratin. Both enzymes were partially inhibited by Ag+ and Hg2+. PMSF completely inhibited the enzymes, while dithiothreitol and 2-mercaptoethanol stimulated their activities, suggesting to be thiol-dependent serine proteases. The enzymes were stable in the presence of the surfactants and bleaching agent (H2O2) and relatively stable in presence of some commercial detergents. PMID- 17419290 TI - Novel yeast cell dehydrogenase activity assay in situ. AB - The aim of this research was to develop a suitable method of succinate dehydrogenase activity assay in situ for different industrial yeast strains. For this purpose different compounds: EDTA, Triton X-100, sodium deoxycholate, digitonin, nystatin and beta-mercaptoethanol were used. The permeabilization process was controlled microscopically by primuline staining. Enzyme assay was conducted in whole yeast cells with Na-succinate as substrate, phenazine methosulfate (PMS) as electron carrier and in the presence one of two different tetrazolium salts: tetrazolium blue chloride (BT) or cyanoditolyl tetrazolium chloride (CTC) reduced during the assay. In comparabile studies of yeast vitality the amount of intracellular ATP was determined according to luciferin/luciferase method. During the succinate dehydrogenase assay in intact yeast cells without permeabilization, BT formazans were partially visualized in the cells, but CTC formazans appeared to be totally extracellular or associated with the plasma membrane. Under these conditions there was no linear relationship between formazan color intensity signal and yeast cell density. From all chemical compounds tested, only digitonin was effective in membrane permeabilization without negative influence on cell morphology. Furthermore, with digitonin treated cells a linear relationship between formazan color intensity signal and yeast cell number was noticed. Significant decreasing of succinate dehydrogenase activity and ATP content were observed during aging of the tested yeast strains. PMID- 17419291 TI - In vitro activity of caspofungin against planktonic and sessile Candida sp. cells. AB - Candida sp. may be regarded as one of the leading etiologic agents of hospital acquired infections, including those related with the indwelling medical devices, which become colonized by the yeasts, accompanied by biofilm formation. In this paper we assayed in vitro susceptibility to caspofungin of planktonic and sessile cells of nasopharyngeal isolates of Candida sp. Two types of biomaterials were used - silicone elastomer-coated latex urinary Foley catheter and PCV Thorax catheter. The minimal inhibitory concentrations (MIC) of caspofungin for planktonic Candida sp. cells ranged from 0.008 to 0.031 mg/l, while the minimal fungicidal concentrations (MFC) from 0.008 to 0.062 mg/l, with MFC/MIC ratios < or = 2. The minimal concentration of caspofungin preventing adhesion process of Candida sp. on both biomaterials ranged from 0.004 to 0.031 mg/l, while preventing biofilm formation from 0.004 to 0.062 mg/l. In contrast, much higher minimal concentrations of caspofungin were needed to eradicate the mature biofilm (0.25 to >8 mg/l). In all cases, drug concentrations depended on the strain and the biomaterial used. Our preliminary data suggest that caspofungin, showing good anti-adherent activity in vitro against Candida sp., appears to be a potential agent rather for prophylaxis of the yeast infections associated with biomaterials but not for their treatment. PMID- 17419292 TI - Enhancement of oil degradation by co-culture of hydrocarbon degrading and biosurfactant producing bacteria. AB - In this study the biodegradation of oil by hydrocarbon degrading Pseudomonas putida in the presence of a biosurfactant-producing bacterium was investigated. The co-culture of test organisms exhibited improved degradation capacities, in a reproducible fashion, in aqueous and soil matrix in comparison to the individual bacterium culture. Results indicate that the in situ biosurfactant production not only resulted in increased emulsification of the oil but also change the adhesion of the hydrocarbon to cell surface of other bacterium. The understanding of interactions beetwen microbes may provide opportunities to further enhancement of contaminants biodegradation by making a suitable blend for bioaugmentation. PMID- 17419293 TI - Biotransformation of phosphogypsum on distillery decoctions (Preliminary results). AB - The paper presents the activity of anaerobic bacterial communities isolated from soil polluted by aircraft fuel on distillery decoctions with phosphogypsum. The microorganisms were selected using the microcosms method, and then enriched on Postgate medium with ethanol. The isolated communities became the inoculum to establish a culture on potato and rye distillery decoctions. The obtained results show that a simultaneous removal of two industrial wastes such as phosphogypsum and distillery decoctions is possible. The introduction of a inoculation comprising a selected anaerobic bacterial community into the culture does not influence the increase of the biotransformation process efficiency. PMID- 17419294 TI - Survival of Proteus mirabilis O3 (S1959), O9 and O18 strains in normal human serum (NHS) correlates with the diversity of their outer membrane proteins (OMPs). AB - Urinary tract infections are frequently caused by Proteus mirabilis strains. In the previous studies there were defined the complete structures of O polysaccharide parts of lipopolysaccharides from strains: P. mirabilis O3 (S1959), P. mirabilis O9 and P. mirabilis O18. In the present study it was investigated bactericidal effect of normal human serum (NHS) to P. mirabilis strains. We also focused on the diversity of outer membrane proteins (OMPs) being separated on a gel isolated from tested strains. Serial passage of P. mirabilis O18 in 90% normal bovine serum (NBS) contributed to over-expressing some classes of OMPs. PMID- 17419295 TI - Lack of an association between Helicobacter infection and autoimmune hepatitis in children. AB - An association between Helicobacter infection and autoimmune hepatitis (AIH) in children was investigated. The prevalence of antibodies to H. pylori did not differ between the AIH and the control group, (22% versus 14%), and antibodies to non-gastric Helicobacter were not detected in either group. H. pylori DNA was found in two AIH liver tissues, but Helicobacter was not cultured from any sample. PMID- 17419296 TI - [The Endocrinology Institute celebrates its 50th anniversary. How is the Institute poised for the next 50 years?]. AB - This paper briefly describes the recent research, therapeutic and pedagogical activities of the Institute of Endocrinology in Prague, which will celebrate the 50th anniversary of its foundation in the beginning of 2007. Current research covers a broad spectrum of topics in endocrinology, which include thyroid and steroid hormone research, neuroendocrinology, immunoendocrinology, molecular endocrinology, endocrinology of aging as well as diabetes and obesity research. Not only clinical studies but also epidemiological and basic research studies are carried out at the institute. Each year approximately 25-30 research projects are conducted, financed both locally (Grant Agency of the Ministry of Health IGA, Grant Agency of the Czech Republic GACR etc.) and by foreign agencies (COST project, 6th Framework program etc.). Since the year 2000, researchers and physicians from the institute have presented 511 papers, of which 162 (32%) were published in internationally impacted journals. Almost 45 000 patients are examined each year at the three clinical departments: the Department of Clinical Endocrinology, the Obesity Management Centre and the Department of Functional Diagnostics. New diagnostic strategies such as assays of recently discovered hormones, clamp techniques, and methods of molecular biology, have been introduced into the regular clinical practice. Recent modernization of the biochemical laboratories and sampling procedures has resulted in modern facilities which offer a broad spectrum of biochemical and hormonal assessments. About 300 patients undergo laboratory investigations daily. Currently, more than half a million biochemical and hormonal assays per year are performed in the institute. The Obesity Management Centre, which was incorporated into the institute in 2002, provides a comprehensive obesity management program focused mainly on the treatment of severely obese patients and patients with high cardiometabolic risks. The institute is involved in pregraduate education at the Faculties of Medicine and at the Faculty of Science of the Charles University and at the University of Southern Bohemia. Both physicians and researchers of the institute contribute significantly to the postgraduate education in endocrinology as the institute is the seat of the Sub-Chair of Endocrinology of the Institute of the Postgraduate Education of Health Care Providers. Both the institute and the Endocrine Sub-Chair obtained the accreditation in endocrinology from the Ministry of Health. Since the acquisition of the Obesity Management Centre, the institute offers a unique one-week training program for obesity specialists organized by the Czech Society for the Study of Obesity. In the forthcoming years closer cooperation with Charles University and the Academy of Sciences is highly likely and should be reflected by a new legislative status of the Institute. PMID- 17419297 TI - [Half century of the continuation of the Endocrinology Institute in Prague]. AB - At the occasion of the 50th anniversary of its foundation, the short history of the Institute of Endocrinology in Prague is outlined. The early years were connected with its founder, Prof. As. MUDr. Karel Silink, DrSc., who carried out an imposing epidemiological survey of iodine deficit in the Czech Republic at the beginning of fifties and who founded the institute on 1. 1. 1957. Not only the thyroid gland, but also the use of radioisotopes, enzymology, steroid hormones and cybernetics were the aims of his interest. The Institute survived the turbulent events of the second half of the 20th century and the successors of K. Silink introduced also endocrine immunology, paediatric endocrinology, molecular biology, neuroendocrinology and obesitology into the normal program of research and health care, which is now-a-days applied to over 150,000 patients. PMID- 17419298 TI - [History, presence and trends of steroid endocrinology]. AB - The millstones achieved in the field of steroid endocrinology since thirties until present are outlined, including major goals obtained in the author's laboratory. After the brief historical survey the following chapters are devoted to steroid laboratory diagnostics, therapy and expected trends. PMID- 17419299 TI - [Study of the genetic causes of polygeneticallv determined endocrinopathies- patience can bring success]. AB - Complex endocrinopathies, such as diabetes mellitus, obesity, polycystic ovary syndrome and osteoporosis belong to the most common diseases but their pathogenesis is still not fully explained. Environmental fadors along with genetic factors contribute to their occurrence and development. The study of genetic background is based on different strategies, mostly on linkage analysis and candidate gene approach. The common forms of these endocrinopathies do not seem to be the result of a defect of one or several major genes but the search for complex gene-gene, gene-environment interactions is needed. The article gives a short review of the recent knowledge together with our own experience in the field of study of the genetic background of polygenic diseases. PMID- 17419300 TI - [Growth hormone treatment in the Institute of Endocrinology]. AB - Institute of Endocrinology became a pediatric center for growth hormone treatment in 1993, after the existing cohort of children treated in the Department of Pediatrics of the Postgraduate Medical School (IPVZ, FTN) had been transferred here. In 2004 it became also a center for adults. At the end of 2006 the total number of those, who underwent the treatment amounted to 408 patients. Their indications come out of internationally observed clinical and laboratory criteria while the treatment itself is regularly controlled in the 3-months periods. It is also a matter of standardized evidence in international protocols. Our insurance covers that financially demanding treatment completely. As the best criterion for the growth hormone efficacy in children we follow the growth velocity, the benefit is however perceived in other clinical and laboratory indices as well. In adults the success criteria are metabolic parameters, the increase of non-fatty body mass, increase of bone density and improvement of subjective quality of life. Our experience in growth hormone treatment is propitious, with 80% of children evaluated as successful. No adverse events has been ever registered; the before time discontinuation of treatment has been decided only four times. Intermittent summer camps, weekend activities and social meetings of patients contribute to the good compliance and collaboration. PMID- 17419301 TI - [The role of complement in autoimmune thyroid disorders]. AB - The complement system is a crucial part of the innate immune system. In systemic autoimmune disorders, its effects tend to be protective. On the contrary, in the autoimmune thyroid disorders (AIT) the complement actively attacks thyrocytes, which express a number of complement components as well as complement inhibitory molecules. According to the experimental studies different ways of complement activation might occur in the thyroid tissue. All ensue via the classical pathway that is started either by immune complexes containing complement activating autoantibodies against thyroid autoantigens, or by direct binding of C4 to the molecule of thyroid peroxidase (TPO); or by direct complement activation by reactive oxygen radicals. Thyrocytes are relatively resistant to the complement attack. However, sublethaly injured thyrocytes release proinflammatory cytokines and reactive oxygen radicals and thus promote the inflammatory process in the thyroid. So far, the clinical significance of the complement in the thyroid has been studied only in postpartum thyroiditis. The exact role of complement in the pathogenesis of AIT remains to be elucidated. PMID- 17419302 TI - [Quality of life in the patients with disorders of sexual development and with Y chromosome in karyotype]. AB - New nomenclature of disorders of sexual development is provided. Contemporary the management and the quality of life in the patients with pure gonadal dysgenesis and mixed gonadal dysgenesis is assessed. PMID- 17419303 TI - [Polycystic ovary syndrome in 2006]. AB - Polycystic ovary syndrome was defined as the combination of anovulation and hyperandrogenaemia (NIH 1990). Another definition used the combination of ultrasonographic appearance of polycystic ovaries and/or anovulation and/or hyperandrogenaemia or cutaneous manifestations of hyperandrogenism (Rotterdam). The population defined according to NIH is probably in greater risk of insulin resistance and obesity. Pathogenesis of PCOS is not clear till now. Dysregulation of ovarian steroidogenesis could be one of the causes of the full-blown syndrome. Up-regulation of steroidogenic enzymes, probably due to the exaggerated expression of transcription factors such as GATA-6 or retinoids could be involved. Insulin sensitisators are now widely used in the therapy. They could beneficially modify not only insulin resistance and dyslipidaemia, but also ovarian and adrenal steroidogenesis. Metformin and glitazones improve anovulation however the studies conducted till now were not representative concerning the point of successful pregnancy. PMID- 17419304 TI - [Neurosteroids and their function]. AB - Neurosteroids are steroid structure hormones with neuroactive function. Neurosteroids have rapid, non-genomic actions in CNS. Non-conjugated metabolites of progesterone such as allopregnanolone, are potent positive modulators of GABAA receptors. They open ion channels for Cl- with analgetic, hypnotic, anxiolytic and anticonvulsant effects. By sulphatation the modulation on GABAA receptors is changed to negative with opposite effect. 19-C-steroids as dehydroepiandrosterone and its sulphate are negative modulators of GABAA receptors acting as an excitant and proconvulsant. They are able to modulate positively N-methyl-D-aspartate (NMDA) receptors and open ion canals for Ca2+. Changed (lowered) neurosteroid levels can be involved in many pathological processes as premenstrual syndrome, stress, depression, some forms of epilepsy, Alzheimer disease etc. Future study targeted on regulation of their production and metabolism and understanding of the mechanism of their actions will help to use them therapeutically. PMID- 17419305 TI - [Regulatory network of transcription factors: their key role in the development and function of pancreas]. AB - Recent molecular biological studies of the pancreatic development have shed new light on the cause of MODY and diabetes mellitus type 2. The identification of transcription factors participating in the regulatory networks cooperating in the development and in adult exocrine and endocrine pancreas enabled the better understanding of the genetic background and subsequent phenotypic features of diabetes. PMID- 17419306 TI - [Genetic background of mitochondrial diabetes]. AB - Diabetes mellitus type 2 represents a heterogenous disease characterized by impaired glucose homeostasis. The disorder clusters in families suggesting genetic disposition, however the mechanism underlying is unknown. Many studies show more frequent maternal transmission of diabetes in the families. One of huge range of explanation is exclusively maternal transmission of mitochondria. Mitochondria are power organelles which produce ATP molecules by oxidation reduction reactions via the respiratory chain. They contain their own genome which codes subunits of the respiratory chain and proteosynthetic apparatus for proteins encoded by this genome. Pathogenic mutations of mitochondrial DNA can affect the activity of the respiratory chain and result in various phenotypes. Mitochondrial diabetes is commonly associated with neuromuscular disorders and often presents with nonautoimmune beta cell failure. Although mitochondrial mutations are associated with diabetes, their low frequency does not explain reported more frequent maternal transmission of diabetes mellitus type 2. PMID- 17419307 TI - [Obesity based on mutation of genes involved in energy balance]. AB - Within the last decade an intensive research led to an identification of several genes which are involved in a regulation of energy balance. In most cases, carriers of these gene mutations do not exhibit further characteristic phenotypic features except for a severe obesity. Obesity based on mutation of one gene product is called monogenic obesity. Mutations in genes for leptin, leptin receptor, proopiomelanocortin, prohormone convertase 1, melanocortin 4 and 3 receptor disrupt the physiological humoral signalization between peripheral signals and the hypothalamic centres of satiety and hunger. Defects of all above mentioned genes lead to phenotype of abnormal eating behaviour followed by a development of severe early-onset obesity. Mutations of melanocortin 4 receptor gene represent the most common cause of monogenic obesity because they are detected in almost 6 % children with early-onset severe obesity. Mutations of the other genes involved in energy homeostasis are very rare. Although these mutations are sporadic we assume that further research of monogenic forms of obesity might lead to our understanding of physiology and pathophysiology of regulation of the energy homeostasis and eating behaviour. Additionally, they may open new approach to the management of eating behaviour and to the treatment of obesity. PMID- 17419308 TI - [Osteoporosis and aterosclerosis--is there any pathogenetic association?]. AB - Fundamental cytokine regulating remodelation of the skeleton is receptor activator of nuclear factor kappa B ligand (RANKL). RANKL is counter regulated by soluble receptor osteoprotegerin (OPG). While RANKL activates osteoclastic bone resorption, the OPG stimulates bone formation. RANKL/OPG system (TRANCE axis) is activated in favour of RANKL in estrogen deficiency, inflammation, bone malignancies and during the treatment with glucocorticoids. TRANCE axis is functional also in other tissues including vessel wall, where dysbalance with superiority of RANKL leads to atherogenesis. Molecules blocking RANKL (specific antibodies and OPG) are potential drugs for treatment of osteoporosis, atherosclerosis, inflammation diseases, myeloma or osteolytic bone metastases. This review is focused on pathogenetic role of TRANCE axis in the development of osteoporosis and atherosclerosis and on its use in diagnosis and treatment of both degenerative diseases. PMID- 17419309 TI - [The polycystic ovary syndrome and its male equivalent]. AB - BACKGROUND: The polycystic ovary syndrome is multifactorial disease. The autosomal genetic transfer of the disease predisposition is the basis for hypothesis that there can exist a male equivalent of PCOS. Androgenetic alopecia has been suggested as the symptom of the male phenotype of PCOS. METHODS AND RESULTS: A group of 30 men with premature hair loss was involved in the present study. In all individuals, their hormonal profile was determined and insulin tolerance test was made. Robust Mann-Whitney test and Fisher's exact test were used for statistic analysis. Based on the laboratory findings two subgroups of individuals were shown. The first one revealed similar hormonal changes as women with PCOS, namely lower SHBG, lower FSH and elevated free androgen index. The other had either no anomaly in steroid spectrum. Both subgroups did not differ in either BMI or age. The subgroup with hormonal changes resembling those of PCOS, showed a significantly higher insulin resistance than the group without these changes. CONCLUSIONS: Based on our results it can be concluded that men with premature alopecia and hormonal changes partially resembling those typical for female PCOS, might probably represent the male equivalent of PCOS. PMID- 17419310 TI - [Polyglandular activation of autoimmnunity as a manifestation of subclinical endocrinopathies]. AB - BACKGROUND: Autoimmune thyropathies belong to the most frequently occurring autoimmune endocrinopathies. Autoimmune thyropathies occur either independently or linked to known polyglandular syndromes of type I-III. METHODS AND RESULTS: During the last decade, we observed a group of patients with autoimmune thyroiditis, in which autoimmune endocrinopathies were mutually associated and named the symptoms of this group "polyglandular activation of autoimmunity". The frequency of the occurrence of autoantibodies against other endocrine organs in this group was determined and the most frequent was the occurrence of the autoantibodies against steroid producing cells - anti-ovary 28 %, anti-adrenal 23 %, anti-testes 12 %. Considering the most frequent occurrence of autoantibodies against steroid producing cells, attention has been paid namely to patients with autoimmune thyroiditis and a concurrent occurrence of anti-adrenal autoantibodies. In the foreground of the clinical picture of these patients were dysregulations on the metabolic and circulation levels together with symptoms of discomfort (subfebrile condition, arthralgia and fatigue). Heavy fatigue of these patients was linked to the changes of levels and mutual ratio of melatonin and serotonin and regarding autoantibodies, mainly autoantibodies against steroid producing cells, namely against the particular cells of the layers of adrenal cortex played a role. CONCLUSIONS: The presence of autoantibodies influenced also the functional response, namely the ACTH/cortisol ratio. Autoantibodies detected were not anti-21-hydroxylase autoantibodies, typical for autoimmune polyglandular syndrome II, but antibodies against antigens of other molecular weight. PMID- 17419311 TI - [Evaluation of results of iodine prophylaxis in Czech Republic]. AB - BACKGROUND: Improvement of system of iodine prophylaxis in Czech Republic resulted in compensation of iodine deficiency. Effects of increased iodine intake on the status of thyroid gland (volume, function, autoimmune disorders) should be determined. The aim of the study was to evaluate those effects in an epidemiological survey. METHODS AND RESULTS: A randomly selected sample of adult population (18-65 years old) from 3 regions was investigated using the same methods at the interval of 5 years. A significant increase of ioduria was proved with the marked increase of prevalence of high category (over 300 ug of I per 100 mL of urine). Volume of the thyroid gland decreased significantly in women, not in men. No significant changes of TSH were observed, however FT4 level slightly, but significantly increased. CONCLUSIONS: Repeated epidemiological survey of the randomly selected sample of adult population in 3 regions verified the effectiveness of improved iodine prophylaxis with not unequivocally positive increase of high (supraphysiological) category of ioduria. Decrease of thyroid volume was proved in women only, a tendency of mild (clinically not important) increase of thyroid function was also observed. PMID- 17419312 TI - [Diabetes mellitus in adult patients with type I diabetes shows immunological, functional and clinical differences depending on the presence of autoimmune thyroiditis]. AB - BACKGROUND: Autoimmune diabetes is usually accompanied by other autoimmune endocrinopathies, most often by autoimmune thyroiditis (AIT), but it is not clear whether diabetes in these patients differs from diabetes without AIT. METHODS AND RESULTS: Eleven-year follow-up of 47 young adult Type 1 diabetic patients with respect to the presence of AIT (group I - positive antibodies against microsomal peroxidase, antiTPO, and tyreoglobulin, antiTgl, group II - only positive antiTPO, group III - without AIT) showed: a) cessation of endogenous insulin secretion (Cpeptide <0.03 nmol/l) in 100 % of patients with AIT (in group I between year 2 and 9, in group II between year 3 and 11, p<0.05), while in group III without AIT only in 55 % of patients (I,II vs. II, p<0.001); b) higher prevalence of antiGAD values > 5 U/ml in group I when compared to patients without AIT (I vs III, p<0.05); c) tendency toward higher doses of insulin needed for diabetes compensation in patients without AIT; d) the highest prevalence of organ-specific and systemic autoantibodies in group I with the most distinct manifestations of AIT, and the lowest prevalence in group III without AIT (statistically significant). CONCLUSIONS: Autoimmune diabetes in adults with AIT compared to diabetes occurring isolated showed differences in the area of autoimmunity against islets of Langerhans, Langerhans islets' function and in the clinical course of the disease. PMID- 17419313 TI - [Endocrine orbitopathy and significance of autoantibodies against 1D protein]. AB - BACKGROUND: Endocrine ophthalmopathy is a chronic eye disease, characterized by inflammation in parabulbar and retrobulbar space, occurring usually in Graves' thyrotoxicosis. Although the pathogenesis of the disease has not been clarified until now, it is accepted that this disease is of an autoimmune nature, where the targets of the autoimmune reaction are the antigens shared by thyroid and orbit tissue. The autoantibodies against recombinant 1D protein are highly specific and sensitive for the diagnosis of endocrine orbitopathy. METHODS AND RESULTS: The aim of our study was to establish, whether the autoantibodies against 1D protein are found predominantly in patients with clinically expressed endocrine orbitopathy. We evaluated in 30 patients with clinically expressed endocrine orbitopathy the thickness of the three retrobulbar eye muscles, damaged by endocrine orbitopathy, determined the parameters of thyroid hormones and anti-TSH receptor autoantibodies. In all patients the detection of circulating autoantibodies against recombinant 1D protein was performed. Autoantibodies against recombinant 1D protein were found in all patients with clinically expressed endocrine orbitopathy. CONCLUSIONS: Immunoreactivity did not depend on the duration or severity of the eye disease, neither on patients' age. We did not find any correlation between the thickness of eye muscles and the titre of anti TSH receptor autoantibodies, levels of ssTSH and free thyroxine and also any correlation between the thickness of eye muscles and the disease duration. PMID- 17419314 TI - [Influence of early postnatal nutrition in preterm infants on their anthropometric and hormonal characteristics at the age of 10 years]. AB - BACKGROUND: [corrected] A protective effect of breast feeding on the development of obesity has been clearly demonstrated. Several studies associated higher protein intake in postnatal period with the earlier "adiposity rebound" and the higher BMI in later life. METHODS AND RESULTS: Twenty eight girls born at 34th week of pregnancy (weight: 2.22+/-0.31 kg, length: 45.2+/-2.5 cm, BMI: 11.0+/-1.5 kg/m(2)), were subdivided in two groups differing in protein intake during the first postnatal month due to either breast feeding or formula feeding (113.5+/ 18.7 g/month vs. 174.00+/-14.3 g/month; p<0.0001). Groups did not differ in average body weight, height and BMI at birth. Anthropometric and hormonal characteristics, body composition, energy and macronutrient intakes were determined at the age of 10 years in girls together with parental BMI. Association between the early postnatal nutrition and anthropometric and hormonal indexes at 10 years was evaluated. No significant differences between the groups in anthropometric and hormonal characteristics were found at 10 years except for IGF-1 which was significantly higher in the group with increased postnatal protein intake. Protein intake during the first postnatal month was not related to fat mass and leptin levels at 10 years. Protein intake during the first postnatal month significantly correlated with IGF-l level, BMI and body circumferences at 10 years. Postnatal fat intake correlates with body circumferences and IGF-1 and also with sagital diameter and leptin at 10 years. CONCLUSIONS: Early postnatal nutrition could encode IGF-1 in later life and this way may be involved in body size programming. Factor analysis revealed IGF-1 as a link between the postnatal energy and macronutrient intake and anthropometric indices and leptin at the age of 10 years. PMID- 17419315 TI - [Relationships between the "eating inventory" factors, socioeconomic status, anthropometric body adiposity indexes and health risks in Czech population]. AB - BACKGROUND: The aim of the study was to compare and evaluate psychological traits of eating behaviour assessed using the Eating Inventory (El) known also as the Three Factor Eating Questionnaire (TFEQ) and their relationship to body adiposity, health and social characteristic in a quota sample of Czech adults. METHODS AND RESULTS: The sample included 1624 women and 1429 men who were interviewed individually by instructed investigators. The sample was quota representative--for gender, age, size of residential location, region and socioeconomic status in Czech adults. Anthropometric data were obtained together with socioeconomic and lifestyle information. Subjects filled out the EI. Health status was evaluated according to the data reported by GPs. Our results show, that women had higher restraint but lower disinhibition and hunger scores than men. Hunger and disinhibition were always strongly associated, whereas restraint was negatively related to the other two factors in men, and to hunger in women. In both men and women an educational level positively correlated with dietary restraint and negatively with hunger. However, significant negative relationship between educational level and disinhibition was revealed only in men. In backward stepwise regression analysis both dietary restraint and disinhibition predicted BMI and waist circumference. BMI and waist circumference were negatively related to restraint, but positively to disinhibition. Restraint and disinhibition were associated with prevalence of comorbidities in men, while disinhibition and hunger were in women. Individuals with high scoring in disinhibition score (upper quartile) exhibited significantly higher prevalence of hypertension, cardiovascular diseases and hyperlipidaemia than those who scored low (lower quartile). These relationships were most marked in middle-aged subjects. CONCLUSIONS: Psychological traits of eating behaviour seem to have an important role in the development and clinical expression of body adiposity. Especially disinhibition is significantly associated with BMI and waist circumference and prevalence of several pathologies. PMID- 17419316 TI - [Levels of hormones in plasma and in synovial fluid of knee joint of patients with rheumatoid arthritis]. AB - BACKGROUND: Dysfunction of endocrine system is very likely one of the important risk factors involved in the pathogenesis of rheumatoid arthritis. The aim of the present study was to investigate the levels of selected hormones in plasma and in synovial fluid of knee joint of patients with rheumatoid arthritis or with osteoarthritis, which could affect the inflammatory processes. METHODS AND RESULTS: Thirty nine patients with rheumatoid arthritis (22 females and 17 males) and 12 patients with osteoarthritis (6 females and 6 males) were investigated. Concentrations of the following hormones were determined in plasma and synovial fluids: cortisol, 17-beta-estradiol, progesterone, dehydroepiandrosterone, aldosterone, testosterone, prolactin, insulin and C-peptide by using radioimmunoassay kits. Increased levels of 17-beta-estradiol and insulin were found in patients with rheumatoid arthritis as compared to those with osteoarthritis. The plasma concentrations of other hormones under study were not significantly different in these groups of patients. Higher levels of 17-beta estradiol, progesterone and aldosterone were noted in inflammatory knee exudate of patients with rheumatoid arthritis. The levels of other hormones in exudates of patients with rheumatoid arthritis and those with osteoarthritis were not significantly different. The ratio of 17-beta estradiol / cortisol, 17-beta estradiol / testosterone and 17-beta estradiol / dehydroepiandrosterone showed increased proportions of estrogens over androgens or glucocorticoids in exudate from patients with rheumatoid arthritis. CONCLUSIONS: These results demonstrated that steroid and peptide hormones are transferred to synovial fluid of knee. The presence of insulin, C-peptide and aldosterone was described for the first time in synovial fluid. In patients with rheumatoid arthritis a predomination of the levels of proinflammatory estrogens over androgens was found in knee exudate. Also the levels of aldosterone and progesterone were elevated in inflammation knee exudate. This suggests that these hormones present in synovial fluid may affect the local rheumatoid inflammatory processes. PMID- 17419317 TI - [Fragments of reminiscence on the Stepanska street]. PMID- 17419318 TI - [Control of the rabbit's IOP after topic instillation of antiglaucomatic latanoprost and amino acid arginine with lysine mixture]. AB - PURPOSE: To evaluate the effect of locally instilled 10% L-arginine.HCI and 10% L lysine.2HC1.2H20 in 0.005% Latanoprost (Xalatane) mixture on the physiological IOP in rabbits. METHODS: After instillation of the arginine and lysine in 0.005% latanoprost mixture into the left conjunctival sac of 5 female rabbits of the New Zealand White species the, IOP was measured at the time of instillation, and 5, 15, 30, 60, 120, 180, 240 min. and 24 hours after the instillation. The right eye was used as the control. RESULTS: The mixture of arginine with lysine in 0.005% latanoprost decreased significantly the IOP value. The decrease had two peaks. The major decrease of the IOP was observed after 30 min. (3.1 mmHg) and after 24 h (5.0 mmHg) after instillation of this mixture. Between 120 min. and to 240 min., the mild decrease of the mean value of the IOP was found at the control eye as well. CONCLUSIONS: The mixture of arginine with lysine in 10 % concentration in 0.005 % latanoprost (Xalatane) decreased significantly the physiological IOP values in rabbits during the whole experiment. The decrease was significant in comparison both to the IOP of the control eye and to the effect of the amino acids arginine and lysine alone. PMID- 17419319 TI - [The use of the toric intraocular lens in treatment of complicated cataract and high degree astigmatism]. AB - The authors refer about highly effective and in our ophthalmology not yet described method of surgical treatment of complicated cataract in patients with high degree of preexisting astigmatism over 3.0 cylindrical diopters. According to the etiology of astigmatism, in the group, there are patients with cataract and high ametropia after previous corneal transplantation and patients with cataract and preexisting high ametropia without any previous surgery. The referred group consists of 14 patients (19 eyes), and 9 of them (9 eyes) underwent in the past penetrating keratoplasty (PK) due to keratoconus at our department. The common sign of this subgroup was unsatisfactory postoperative refraction and later on the progressive cataract development. The other part of the group consisted of 5 patients (5 eyes) and they underwent no surgery in the past. For the surgical treatment of cataract and astigmatism at once we have used the individually manufactured silicone toric intraocular lens made by the company Human Optics (type MS 6116 TU). The average age of the whole group at the time of the cataract surgery was 51.5 +/- 6.8 years (range, 39-66 years). The group consisted of 8 women (57.1%) and 6 men (42.9%). The average duration of the period between the keratoplasty and the cataract surgery was 4.8 +/- 3.6 years. The uncorrected visual acuity before the cataract surgery was 0.10 +/- 0.11, and the best-corrected visual acuity was 0.39 +/- 0.22. The average value of the preoperative refractive error was +3.4 +/- 5.0 dioptres (range, -11.0 to +10.0 dioptres) and -6.4 +/- 2.3 cylindrical dioptres (range, -12.0 to -3.0 cylindical dioptres). The average follow up period was 9.6 months (range, 4-18 months). In the postoperative period we noticed improving of the uncorrected visual acuity to the value 0.50 +/- 0.26, and the best-corrected visual acuity improved finally to 0.56 +/- 0.26. The final average refractive error decreased to 0.44 +/- 0.86 dioptres and -1.8 +/- 0.77 cylindrical dioptres. PMID- 17419320 TI - [Intraoperative floppy iris syndrome]. AB - In the year 2005, Chang and Cambell described unusual reaction of the iris during the cataract surgery in patients treated with tamsulosine. This was named as IFIS, an acronym for the Intraoperative Floppy Iris Syndrome. In its advanced stage, the syndrome is characterized by insufficient mydfiasis before the surgery, narrowing of the pupil during the surgery, its impossible dilatation during the surgery by means of stretching, unusual elasticity of the pupilar margin, surging and fluttering iris with tendency to prolapse. The same manifestations we observed in our patients and we confirm the direct connection with tamsulosine hydrochloride treatment. Tamsulosine is the antagonist of alpha 1A adrenergic receptors whose are present, except in the smooth musculature of the prostate gland and the urinary bladder, in the iris dilator as well. At the same time we observed this syndrome rarely in some patients not using tamsulosine. In most cases, these patients were treated with antipsychotic drugs. PMID- 17419321 TI - [Functional examination of retinal vessels in patients with central retinal vein occlusion]. AB - Retina vessel analyzer (RVA) provides the functional examination of retinal vessels based on the analysis of the extent (size) of their dilation and constriction. The RVA measures continuously on-line the diameter of retinal arteries and veins after different kind of stimulation. Beyond dynamic vessel analysis, another possibility of the RVA's utilization is a static vessel assessment, measuring the arterial and venous diameter ratio (A/V ratio), which provides the information about the rate of arterial vasoconstriction. The aim of the presented study was to investigate static and dynamic retinal vessel changes in patients with central retinal vein occlusion (Group 1). The second investigated group consists of patients with arterial hypertension; as a control group, healthy persons without any vascular disease were examined. Altogether 40 eyes were examined. Statistically significant differences of A/V ratio were observed in the static vessel analysis in all three investigated groups. The dynamic analysis showed statistically significant differences in arterial dilatation and constriction between all investigated groups as well. The presented results confirm that the degree of retinal vessels endothelial dysfunction is one of the determinating ethiopathological factors of central retinal vein occlusion. PMID- 17419322 TI - [The influence of LASIK to the retinal nerve fiber layer in myopia]. AB - Purpose of this paper was to evaluate statistically the thickness of the retinal nerve fiber layer (RNFL) measured by means of the GDx analyzer in middle and high degrees of myopia before laser assisted in situ mileusis (LASIK) and in the postoperative period of 6 months. MATERIAL AND METHODS: The group consisted of 35 eyes of 18 patients (8 men and 10 women), the average age was 28.9 +/- 5.08 years of age. The refractive error was from -3.25 dioptres (D) to -11.5 D (average -5.5 +/- 1.4 D). The patients underwent the corrective refractive procedure by means of LASIK to correct the myopia. The thickness of RNFL was measured by means of GDx analyzer with Variable Corneal Compensator before the refractive procedure and 3 and 6 months after this. Before each measurement, a new compensation of the cornea according to the actual refractive status was used. The RNFL thickness values (in microm) were compared and statistically evaluated using of the Wilcoxon's nonparametric pair test in the whole peripapilary ellipse area (TSNIT) and in the superior and inferior quadrants of this ellipse. RESULTS: Statistically significant difference of the RNFL thickness at the 5% level of significance was found in the TSNIT area after 3 months and after LASIK. Statistically significant difference of the RNFL thickness at the 1% level of significance was found in the superior quadrant after 3 months and in the inferior quadrant after 3 and 6 months after LASIK. PMID- 17419323 TI - [The cooperation between the ophthalmologist and the endocrinologist in the treatment of the endocrine orbitopathy]. AB - Authors of this study emphasize the requirement of the cooperation between the ophthalmologist and the endocrinologist in diagnostics and treatment of moderate and severe forms of endocrine orbitopathy (EO). Examinations necessary for diagnosis and possibilities of the systemic treatment are reported. Twenty patients within the group of 70 patients with EO, who had severe form of disease and underwent different combinations of corticosteroid therapy, immunosuppressive therapy, radiotherapy (RA) and orbital decompression were followed up. Authors recommend a dosage of Methylprednisolon (7-9 g) divided into pulses of 1000mg followed by pulses of 500 mg given during 3 to 4 weeks (2-3 infusions per week). They recommend administering Prednison in 60-90 mg doses per day depending on weight of a patient. After daily maximum dose during the first two weeks, the authors recommend to decrease gradually the dose with the total treating period of minimum of a half a year. Decrease of visual acuity depending on EO appeared by 7 patients. It has been stabilized in 6 patients after the treatment of EO. Hand movement remained in one patient with severe neuropathy in spite of urgent orbital decompression. The intraocular pressure has been stabilized in 16 patients after treatment of EO (six patients do not require further antiglaucomatic therapy). The decrease of protrusion occurred in 8 patients after corticosteroid therapy (1-5 mm) and in 5 patients after orbital decompression (6 10 mm). Severe adverse events (herpetic infection, osteoporosis, steroid DM) were reported in 3 patients after repeated courses of corticosteroid therapy. Authors recommend early administration of intensive systemic corticosteroid therapy in active stage of the moderate forms of EO. PMID- 17419324 TI - [Orbital apex syndrome of the aspergillus etiology--a case report]. AB - The authors present a case report of a patient, in whom after a head injury the monolateral blindness occurred. Because of autoimmune thrombocytopeny the patient was treated with long-term corticosteroids. The clinical findings corresponded with the orbital apex syndrome. According to the results of the CT and MRI examinations, the sphenoidotomy was indicated, and the histological findings verified fragments of paranasal sinuses' aspergiloma. During the next course of the disease, despite antimycotic therapy, the progression of the aspergiloma in to the anterior cranial fossa occurred. Invasive sino-orbital aspergilosis, after the penetration of the infectious agent across the wall of the sinus, may cause the orbital apex syndrome with paralysis of all three cranial nerves innervating the extraocular muscles, sensoric defect in the area of the ophthalmic nerve and the involvement of the optic nerve. PMID- 17419325 TI - [Geoinformation technologies in phthisiology]. AB - Special geoinformation technologies (GIS), a new method for the pictorial data presentation as space-time ratios, have emerged in the past decade. The paper considers the theory of geoinformation analysis and the possibilities of applying GIS for tuberculosis monitoring. GIS may be effectively used in phthisiology only provided that the routine procedures for tuberculosis detection are practiced widely. GIS analysis may assist in detecting tuberculosis when contacts cannot be monitored in the usual fashion. Its results may be used to correct tuberculosis controlling measures towards their point application. Extended studies can answer to what extent, the efficiency of tuberculosis-controlling measure will increase when they are used in the most possible tuberculosis transmission areas determined by the GIS techniques. PMID- 17419326 TI - [Use of discriminant analysis for the integrated estimation of the activity of an epidemic process of tuberculosis and the efficiency of medical measures for its prevention]. AB - Whether the discriminant analysis might be used for the integrated estimation of an epidemic process of tuberculosis and for the efficiency of medical measures at the interregional levels was studied. Out of the 8 earlier chosen epidemiological criteria, this analysis could identify 6 most important ones that yielded two canonically discriminant variables. Calculations indicated that over 90% of the whole dispersion of regions in the activity of an epidemic process of tuberculosis might be accounted for by Discriminant Variable One. Variable Two played an indirect role. Out of the 8 clinical criteria, repeated analysis could choose four ones that gave rise to one canonical discriminant variable. The latter provided an integrated estimation of the efficiency of medical measures. The accuracy of prediction of the belonging of regions to high and low epidemic intensity groups was 100%. The findings open up possibilities of designing a present-day tuberculosis epidemic surveillance system at the interregional and regional level. PMID- 17419327 TI - [General principles in the detection of patients and verification of a diagnosis in tuberculous pleurisy]. AB - How detection of patients with tuberculous pleurisy is organized in the Voronezh Region in the period of 1979 to 2003 was studied. Tuberculous pleurisy was diagnosed in 2681 (31.73%) of 8450 patients with pleural effusion of diverse etiology. A package of necessary studies was determined for the verification of the diagnosis. It is suggested that it is expedient to establish a pleural pathology unit at a tuberculosis hospital for enhancement of the efficiency of diagnosis and verification of tuberculous pleuritis. PMID- 17419328 TI - [Comparative characteristics of disseminated pulmonary tuberculosis in patients isolating Mycobacterium tuberculosis with a high and low viability]. AB - The specific features of disseminated pulmonary tuberculosis were studied in patients isolating Mycobacterium tuberculosis (MBT) with a high (n = 135) and low (n = 50) viability. The degree of the pathogen's viability was shown to determine the clinical characteristics of a specific process and the efficiency of its treatment. It is speculated that there is a direct relationship between the viability ofmycobacteria and their virulence. There are differences in the nature of primary drug resistance in MBT with their varying variability. PMID- 17419329 TI - [Interdepartmental interaction in the detection and treatment of patients with tuberculosis]. AB - The experience of interdepartmental interaction in solving the problem of tuberculosis in the Kemerovo Region has been analyzed. The region's administration has strengthened the material-and-technical basis of the service. The advisory tuberculosis council has been set up. Territorial interdepartmental commissions have been organized. The level of an intervention with the general medical network (GMN) has been increased. A qualitative sputum test has been carried out for acid-resistant mycobacteria. The Main Board of the Federal Penal Jurisdiction Service (MBFPJS) and the Board of Internal Affairs are attracted to examine social risk-group persons. Strictly uncontrolled chemotherapy for patients with tuberculosis has been introduced on the basis of GMN at the stage of aftercare and by the forces of the Red Cross Committee. Along with the MBFPJS, continuity has been established in the treatment of patients with tuberculosis. Juridical attraction of bacteria-isolating person to treatment has been introduced. The work done has substantially improved the detection and treatment of patients, but there are problems to be solved. PMID- 17419330 TI - [Mononuclear cell response to fetoproteins in patients with pulmonary tuberculosis in dysregenerative bronchial epithelial changes]. AB - A relationship was studied between the response of mononuclear cells (MNC) to fetoproteins (FP) and the bronchial changes in patients with pulmonary tuberculosis. The response was evaluated by the change in the relative count of CD8+ lymphocytes after MNC incubation with FP as compared with the controls. The value of the response (in scores) was compared with the pathohistological pattern of the bronchial epithelium and interstitial tissue. The studies show that the values of a MNC response to FP are criteria for the presence and absence of dysplasia as one of the types of bronchial epithelial atypia. It is not improbable that patients with chronic inflammatory processes in the lung in the presence of a positive MNC response to FP should be regarded as persons at risk for malignancies. PMID- 17419331 TI - [An association of the reduction in the count of T1 cells in the peripheral blood of patients with pulmonary tuberculosis with the lower monocytic expression of costimulatory CD80 molecules]. AB - The percentage of T1 lymphocytes in the presence of the unchanged content of T2 cells were shown to be decreased in the peripheral blood of patients with pulmonary tuberculosis as compared with apparently healthy donors. In addition, the patients were found to have a significantly reduced quantity of CD80 expressing monocytes, the relative quantities of Th1 and Tc1 cells were directly related to the count of CD80+ monocytes. It is suggested that decreased CD80+ cells is a cause of ineffective antigen presentation that results in a systemic reduction in the count of CD4+ and CD8+ cells producing T1 cytokines and in impairment of an effective specific T1 response. PMID- 17419332 TI - [Mucociliary transport in patients with chronic obstructive lung disease from the cotton-growing areas of Uzbekistan]. AB - Based on the data on 98 patients with chronic obstructive lung disease (COLD) of varying severity from the cotton-growing areas of Uzbekistan, including 66 patients with pesticide accumulation and 32 without the latter, the authors studied the mycociliary transport (MCT) by the time of MCT, adhesiveness, and surface tension of sputum, as well as its content of bound and free water. The studies point to the development of significant mucociliary insufficiency that deteriorates the signs of bronchial obstruction in patients with COLD with pesticide accumulation. With the longer release of an inhaled indicator (MCT time > 48 hours), peak forced expiratory flow rates (PFEFR) were found to considerably decrease in this category of patients than in patients without impaired MCT (PEFR 1.8 +/- 0.6 l/sec versus 5.9 +/- 0.3 l/sec. The increases in sputum adhesiveness and surface tension may be used as an additional criterion of MCT impairments. PMID- 17419333 TI - [Reduced mycobacterial resistance to antituberculous drugs in the experiment and clinic: immediate and long-term results]. AB - A suspension of multidrug resistant clinical Mycobacterium tuberculosis (MBT) strain, at a concentration of 1 x 10(8) microbes per ml, resistant to streptomycin (S), rifampicin (R), isoniazid (I), and kanamycin (K), was in vitro treated for 60 minutes with dissolved ozone (pO3) at a concentration of 0.5-4 microg/ml). Then it was placed in the Lowenstein-Jensen media containing various concentrations of S, I, R, and K. Following 3 months, drug susceptibility was determined by the number of cultured colonies and MBT was used to prepare a suspension at the same concentration, which was again treated with pO3 by the same procedure and placed to the media containing the drugs. A session was thrice repeated. After each pO3 treatment, MBT resistance to I decreased and it completely disappeared after triple treatment. Each pO3 treatment caused a reduction in MBT resistance to R, but it was high (640 microg/ml). After double pO3 treatment MBT resistance to S decreased, but it was recovered after its third ozone treatment. All pO3-untreated control cultures showed a growth of more than 100 colonies. Sixty-eight BALC/s mice were in vivo inoculated via intravenous injections of the clinical MBT strain resistant to S, I, R, and K. The mice were divided into 5 groups: 1) intact mice; 2) those inoculated and untreated; 3) those treated with 1; 4) those treated with I and peritoneally given pO3, 0.5-4 microg/ml); and 5) those given pO3. The animals began dying at month 4 of inoculation. By month 5, all mice, other than intact and pO3-treated ones, died. Passage of MBT from the month by month 4 showed a reduction in their resistance to I in the groups treated with pO3. When the mice were treated with I alone, damages to their livers and spleens were greater than when they were untreated. With co-administration of I and pO3, the damage was least. Treatment provoked a rapid change of MBT to granular and L-forms and MBT was undetectable in its typical form after 1-2-month therapy. The altered MBT formed an untypical histological pattern of tuberculous inflammation in mice in the presence of characteristic cellular cooperation. Clinical studies indicated that 1-6-month concurrent use of chemotherapy and pO3 in patients with drug-resistant tuberculosis eliminated drug resistance of isolated MBT to one of the drugs (I, R, K) in 97.3%, MBT became at once susceptible to I, R, and K in 47.2%. I and/or R were successfully used in the treatment of more than a third of the patients. The studies have demonstrated that most patients with drug-resistant pulmonary tuberculosis can be treated with the most effective drugs provided that systemic intravenous injection of pO3 is made. PMID- 17419334 TI - [Study of a role of artificial mycobacterial particles in the realization of immunological processes in experimental animal tuberculosis]. AB - The protective properties of artificial mycobacterial particles versus BCG vaccine were studied in laboratory animals with experimental tuberculosis. The findings of the decreased rate of a tuberculous process and on the increased mean life span in animals inoculated with M. bovis suggest that immunization of guinea pigs with mycobacterial particles promotes the enhanced development of antituberculous immunity in the animals. The paper proposes a promising method for designing artificial immunogens, the high-polymer antigenic structures that imitates mycobacterial particles. PMID- 17419335 TI - [Investigation of the thermal resistance of Mycobacteria tuberculosis]. AB - Incubation by means of the growth-promoting factor VCG of autoclaved culture broth of M. bovis 8 and tuberculins on the nutrient medium VCG gave rise to microorganisms that could partially restore acid resistance. The isolates reacted with antiserum to M. bovis Vallee in the agglutination test and enzyme immunoassay, which in combination with polymerase chain reaction data verifies their genetic relationship with the pathogen of tuberculosis. The isolates did not show a significant pathogenicity, but they long persisted in guinea-pigs and, in some cases, induced delayed hypersensitivity to tuberculin. When exposed to high temperatures, the pathogen of tuberculosis is presumably to form structures that are capable to restore viability under certain conditions (for example, on the VCG medium). PMID- 17419336 TI - [Implication of specialized sanatoria in the improvement of care to patients with ocular tuberculosis at the present stage]. AB - A total of 6230 case histories and the data of annual reports over the past 7 years were examined. A therapeutic-and-diagnostic process continues to be improved under the present-day conditions. Patients admitted to the sanatorium for the inadequately substantiated etiology of ocular tuberculosis underwent an additional differential diagnostic study. The diagnosis of tuberculosis was verified only in 31.29% of cases. The optimized procedure for etiotropic chemotherapy for patients with new-onset ocular tuberculosis has been introduced. A scheme for intensifying the treatment of recurrent ocular tuberculosis, which permits high therapeutic results to be achieved, has been proposed. It is expedient to reorganize the sanatorium "Red Bank" as one of the sanatorium centers of therapeutic-and-rehabilitative and differential diagnostic care to patients with ocular tuberculosis. PMID- 17419337 TI - [From the history of tuberculosis control in Yakutia (1945-1965)]. PMID- 17419338 TI - [Development of optical introscopy in medical engineering]. AB - The basic features of the process of development of medical optical introscopes are considered. Design characteristics of preproduction and production models of devices for diagnosis of the brain and the mammary gland are described. PMID- 17419339 TI - [An experimental defibrillator with programmable pulse shape]. AB - Results of development of an experimental external electric defibrillator with programmable pulse shape are described. The defibrillator can be used for experimental determination of the optimal shape of defibrillation pulse. PMID- 17419340 TI - [Testing of rhythm identification algorithms for automatic defibrillators]. AB - Testing of rhythm identification algorithms used in automatic defibrillators is required by safety standards. The safety and efficiency of automatic defibrillators depends to a considerable extent on the adequacy of the testing procedure. It is recommended to use annotated public databases of ECG rhythms for testing of rhythm identification algorithms. An effective testing procedure includes two stages: computer simulation and real-time testing of the defibrillator. PMID- 17419341 TI - [Software for polygraphs used in scientific research]. AB - Polygraphs--devices for simultaneous detection of several physiological parameters--are currently used in medicine in psychology. Special polygraphs are also used in scientific research. The process of data acquisition and processing in such polygraphs is usually computer-controlled. The goal of this work was to describe software for detection of polygraph signals used in cardiorespiratory system research. The software supports the experiment script language, allows hyperventilation testing, and makes it possible for the examination subject to control the data detection process. PMID- 17419342 TI - [Classification of human sleep stages based on EEG processing using hidden Markov models]. AB - The goal of this work was to describe an automated system for classification of human sleep stages. Classification of sleep stages is an important problem of diagnosis and treatment of human sleep disorders. The developed classification method is based on calculation of characteristics of the main sleep rhythms. It uses hidden Markov models. The method is highly accurate and provides reliable identification of the main stages of sleep. The results of automatic classification are in good agreement with the results of sleep stage identification performed by an expert somnologist using Rechtschaffen and Kales rules. This substantiates the applicability of the developed classification system to clinical diagnosis. PMID- 17419343 TI - [A P300-based brain-computer interface]. AB - The goal of this work was to describe a system for real-time typing controlled by brain biopotential signals. A 6 ( 6 matrix containing Russian alphabet letters and auxiliary symbols was shown on PC screen. Electroencephalogram was taken, and the P300 component was extracted (this component appeared only upon presentation of a significant stimulus). A combination of several detection methods was used to identify the P300 component, which made it possible to increase the probability of correct identification to 91.6 (5.2%. It was shown that the developed interface could be implemented on the basis of a single active electrode in Pz (Cz) position. PMID- 17419344 TI - [Use of pitch and formant analysis in speech biometry]. AB - The goal of this work was to study the possibility of use of speech biometry for identification of speaker, determination of speech accent, and estimation of the emotional state of the speaker. It was suggested to solve these problems by analysis of the two main components of the linear model of speech production: pitch and formant frequencies. It was shown that pitch and formant analysis provided estimation of the emotional state of the speaker. It was found that sthenic emotions were accompanied by an increase in the pitch and its dispersion. The speech accent was determined using formant analysis and neural network classifiers. Speaker identification was successfully performed in a test group of 50 subjects. PMID- 17419345 TI - [Monitoring of the concentration of lithium and heavy metals in drinking water by the method of stripping voltammetry]. AB - A method for monitoring of the concentration of lithium, zinc, cadmium, lead, and copper in drinking water is suggested. Monitoring can be performed within the range of 1.5(10(-8) - 2.0(10(-6) mg/dm3. A new design of the electrochemical cell is suggested. Analysis is performed against the background of 0.02 M dimethylformamide solution of (C4H9)4NCIO4. The time and potential of electrolysis are determined experimentally. The method can be used in medical research. PMID- 17419346 TI - [Noninvasive methods for blood glucose measurement]. AB - For a long time, blood and urine analysis were the only methods for determination of the concentration of glucose in blood. However, these methods do not provide all necessary information about the state of the disease. Besides, use of these methods can be inconvenient. Thus, the problem of noninvasive diagnosis of diabetes mellitus becomes especially important. In this work, currently available methods for blood glucose measurement are reviewed. PMID- 17419347 TI - [Fever as a very important symptom of infectious and some non-infectious diseases]. AB - Fever is a protective and adaptive reaction, an early and obligatory symptom of infectious diseases in the first place. Historically, this term has been used in the names of more than 20 infectious diseases. Fever may also be a manifestation of other diseases accompanied by aseptic inflammation and neurohumoral disturbances. The growth rate of fever and the character of its circadian fluctuations at the beginning of the disease, as well as its total duration, the height and type of temperature curve, and time of onset of organic lesions should be taken into consideration when performing differential diagnostics of fevers of different origins. The authors developed an algorithm of differential diagnostics of febrile conditions. When an infectious process is excluded reasonably, the patient should undergo further examination for early detection of other diseases accompanied by fever. PMID- 17419348 TI - [Contemporary therapy of chronic obstructive pulmonary disease and its new directions]. AB - Chronic obstructive pulmonary disease (COPD) gives rise to substantial structural changes in the respiratory tract and lungs with the development of chronic inflammation and bronchiolitis, as well as pulmonary parenchymatous destruction and the formation of emphysema. The listed changes substantiate the development of new treatment techniques, which will help correct structural changes in the respiratory tract and will slow down or even reverse progressive worsening of the respiratory function. In the future, the development of blood and/or saliva infection of inflammation biomarkers (e.g. endogenic protease inhibitors, antioxidants, cytokines etc.) will make it possible to find valuable diagnostic signs of forthcoming exacerbation, as well as to evaluate the effectiveness of new therapeutic techniques in clinical studies. COPD, or chronic bronchitis, airway obstruction, and emphysema, should be considered among pathological processes which result in smoking-induced chronic system inflammatory syndrome including cardiovascular diseases, metabolic disorder, and cancer. Possibly, treatment with new preparations, directed towards treating the patient as a whole rather than separate COPD symptoms, will be developed in the future. PMID- 17419349 TI - [Chronic pancreatitis: the modern state of the problem. Part 1. Definition, prevalence, and the issues of ethiology and pathogenesis]. AB - The review presents the latest data on the epidemiology of chronic pancreatitis (CP), its etiology and pathogenesis, and the modern classification of the disease. The review supports the great diversity of etiological factors of CP, which determine peculiarities of its pathogenesis and clinical picture. PMID- 17419350 TI - [Principles of concervative therapy of urolithiasis and prevention of stone formation recurrence]. AB - The modern view of treatment for various forms of nephrolithiasis reflects the main features of nephrolithiasis pathogenesis, but changes in metabolic condition, the presence of dysmetabolism, and the course of pathologic process are not taken into account. Considering that the disease is easier to prevent than to treat, it would be more appropriate to base preventive measures not only on the data from general clinical and biochemical examination, but also on the results of aggregatometry and complex chromato-mass-spectrometric examination allowing prediction of the development of the pathologic process. PMID- 17419351 TI - [Endothelial dysfunction in patients with chronic heart failure, and possibilities of its correction with isosorbide-5-mononitrates]. AB - Vascular endothelial dysfunction has been proven to play an important role in the development and progression of chronic heart failure (CHF). The influence of isosorbide-5-mononitrate monocinque on endothelial function in patients with NYHA II class chronic heart failure complicating the course of coronary artery disease (CAD) with II--III class stable exertional stenocardia was studied. Disturbances in endothelium-dependent vasodilation in a form of a lowered endothelial response to an increase in shift tension during reactive hyperthermia test were noted in patients with CHF complicating CAD. After 12 weeks of treatment with monocinque endothelium-dependent vasodilation normalized, which manifested by an increase in the vasodilating response of the brachial artery to stimulation with an increased flow from 8.2 +/- 1.3 to 11.1 +/- 1.0%. Monocinque administered in a day dose of 40 mg possessed antianginal activity in 85.7% of patients: the frequency of anginal attacks lowered by 59.3%, morphofunctional heart parameters improved which manifested by a 8.7% increase in ejection fraction. Side effects of monocinque, typical of nitrates, were noted in 17.9% of patients and did not require discontinuation of the therapy. PMID- 17419352 TI - [On the issue of the effects of geomagnetic and meteorological activity on patients with arterial hypertension]. AB - The influence of "space weather" (effects of geomagnetic and meteorological activity) on the effectiveness of antihypertensive therapy in patients with arterial hypertension was studied. Blood pressure monitoring using self-control technology was conducted during 12 weeks in 33 patients with I to II degree arterial hypertension receiving different antihypertensive agents in Myasnikov Cardiology Research Institute. Two groups of patients were distinguished according to the effectiveness of the therapy; groups of patients who had two- to four-week periods of spontaneous rhythmical variations of arterial pressure and/or an eluding hypotensive effect, were found as well. Matching these data to the data on geomagnetic activity as well as atmospheric pressure and temperature revealed magnetosensitive patients in the group with multipronged structural and functional changes in target organs requiring combined therapy. The effects of spontaneous rhythmical variations in blood pressure and the effects of antihypertensive therapy elusion, including those in the group of easier-to-treat patients, also correlated with geomagnetic and meteorological activity. These pilot studies demonstrated the necessity to take space weather effects into account when examining and treating cardiological patients, as well as the fact that further research in this direction is needed to clarify what algorithms will be necessary for this purpose. PMID- 17419353 TI - [Hypodynamia and spontaneous venous thromboses]. AB - Data on the prevalence of hypodynamia among individuals with the first and second episodes of spontaneous venous thrombosis in a population sample of Moscow residents are compared. It is stressed that changes in behavioral stereotype, which determine the reduction of physical activity level before the first episode, are significant for the risk of rethrombosis. Besides, the authors discuss a range of questions concerning terminology of low physical activity, distinguishing the reasons for hypodynamia, and evaluation of existing questionnaires for measuring the levels of physical activity. PMID- 17419354 TI - [Hemorheology and hemostasis in patients suffering from ischemic cerebral stroke and metabolic syndrome]. AB - A high level of mortality and disability makes study of various aspects of ischemic cerebral stroke (ICS) an extremely important problem. Presently, combination of various cardiovascular risk factors significantly increases the probability of such life-threatening conditions as ICS. Such risk factors as arterial hypertension, dyslipoproteinemia, excessive body weight, glucose intolerance or diabetes are often combined, which led to a suggestion to consider them together as one symptom complex, metabolic syndrome (MS). It is evident that MS affects the functioning of different organs and systems, including the systems of hemorheology and hemostasis. These are changes in hemorheological and hemostatic parameters that play a leading (in some cases decisive) role in the development of ICS. The present study found that the presence of MS in ICS patients hindered improvement in hemorheological and hemostatic parameters in the course of the disease. This was manifested by the absence of positive changes in thrombocyte aggregation, as well as fibrinogen level elevation in patients with MS in the course of treatment. Elevated blood level of D-dimers within the whole acute period in most MS patients also reflected more active thrombus formation. The development of ICS is very unfavorable to the condition of vascular wall athrombogenic activity in all patients. At the same time, worsening in all the chains of endothelial wall athrombogenic ability was more prominent in MS patients. Significant influence of MS on the forming of prothrombotic condition, which determines unfavorable clinical course of ICS, was demonstrated. PMID- 17419355 TI - [The clinicomorphological characterization of segmental lesion in ulcerous colitis]. AB - The authors analyzed the clinical picture in 18 patients with distal lesion of the colon and an inflammatory focus in the cecum (segmentary lesion), and five patients with distal lesion of the colon revealed with endoscopy and pathomorphological signs of inflammation in visually intact cecum. The analysis shows that in a range of cases the extent of lesion in ulcerous colitis (UC) cannot be determined correctly with colonoscopy. An apparent segmentary character of lesion (inflammation in the distal colon and an inflammatory focus in the cecum) is an additional criterion of unfavorable prognosis of transformation into disseminated UC forms. Such patients need application of the same treatment regimens as in patients with disseminated UC forms. PMID- 17419356 TI - [The peculiarities of the clinical picture and the effectiveness of antiviral therapy of chronic hepatitis related to alcohol consumption plus hepatitis C virus effect]. AB - The purpose of this study was to describe the features of chronic hepatitis when the damaging action of alcohol was combined with that of hepatitis C virus, and to investigate the effectiveness of combined antiviral therapy in this category of patients. The study showed that patients with chronic hepatitis of combined ethiology (alcohol plus HCV) had higher biochemical and histological activity of the disease, as well as certain histological features. The effectiveness of combined antiviral therapy was comparable in both alcohol consumers and non consumers with hepatitis C, which makes it possible to consider the former category as candidates for antiviral treatment under the condition that they follow doctor's recommendations and abstain from alcohol consumption. PMID- 17419357 TI - [Pharmacotherapeutical features of arterial hypertension in patients with diabetes mellitus]. AB - Diabetes mellitus (DM) is often combined with arterial hypertension, which increases the risk of serious complications. The purpose of this review was analysis of the peculiarities of pharmacotherapy with drugs belonging to different groups (ACE inhibitors, calcium channel blockers, alpha- and beta adrenoblockers, diuretics, angiotensin II blockers, and imidazoline receptor agonists) as a part of combined therapy of compensated DM. PMID- 17419358 TI - [The influence of CYP2C9 genetic polymorphism on the pharmacokinetics and pharmacodynamics of warfarin in patients with constant atrial fibrillation]. AB - CYP2C9 is the main enzyme participating in warfarin metabolism, of which genetic polymorphism is typical. The aim of the study was to investigate the influence of having allelic variants CYP2C9*2 and CYP2C9*3 on the pharmacokinetics, dosage regimen, and the rate of hemorrhage in patients with constant atrial fibrillation taking warfarin. Eighty-two patients with constant atrial fibrillation taking warfarin were included in the study. It was shown that in patients with CYP2C9*2 and CYP2C9*3 the clearance of warfarin and its dose were lower, while the episodes of excessive hypocoagulation and hemorrhage associated with warfarin were more frequent than in patients without these allelic variants. Basing on the results of the study, the authors propose an algorithm of choosing the initial warfarin dose depending on CYP2C9 genotype. PMID- 17419359 TI - [Some features of coronary artery disease combined with vertebrogenic thoracoalgias]. AB - The subjects of the study were 4223 patients who were examined for heartache between 1998 and 2005. The patients were divided into three groups. Group I were 617 patients with isolated vertebrogenous cardiomyalgia; group II included 1238 patients with coronary heart disease; group III consisted of 2368 who suffered from both conditions. On the basis of clinicoinstrumental examination differential diagnostic criteria were derived. PMID- 17419360 TI - [Contemporary approaches to evaluation of psychosomatic status in patients with alimentary tract diseases]. AB - The authors present the results of psychosomatic status evaluation performed in 159 patients with biliary dyskinesia, chronic gastritis, and somatoform disturbances. The examination was done using questionnaires, scales, and computer methods. Computer diagnostic methods were proved to be most effective and allowed for quantitative evaluation of psychosomatic disturbances in patients with gastrointestinal diseases. PMID- 17419361 TI - [Bad breath: causes, diagnostic and therapeutic practice]. AB - Bad breath (BB) can be noted in various diseases of oral cavity, nasopharynx, upper gastrointestinal tract divisions (foetor ex ore) or be felt in expired air (halitosis) in various bronchopulmonary diseases, renal or hepatic failure, and diabetic ketoacidosis. BB requires interaction between different specialists making it possible to find the cause of this clinical symptom and the optimal therapeutic approach. PMID- 17419362 TI - [A focal infiltration in the upper lobe of the right lung as a manifestation of the initial stage of secondary syphilis]. AB - Among sexually transmitted diseases, the incidence of primary and secondary syphilis has substantially increased in Russian Federation within the recent years. Modern clinicians, acquainted only with conventional dermatovenerologic symptoms of the disease, are less aware of rare manifestations of syphilis as a generalized infection. A rare case of syphilis with infiltrative pulmonary lesion is discussed in the article; the attention of practitioners is drawn to "unusual" manifestations of syphilis. PMID- 17419363 TI - [Relevance of testicular microlithiasis to the testicular carcinoma in situ in the contralateral testicle]. AB - PURPOSE: To determine the prevalence of testicular microlithiasis (TM) in the contralateral testis of patients with unilateral testicular tumor, and evaluate the association of TM with carcinoma in situ (CIS) MATERIALS AND METHODS: In a period of 12 years, ultrasonography was performed in 41 patients with unilateral testicular tumor. TM was defined as multiple small hyperechogenic foci. Only images of the tumor-free testis were reviewed. In addition, archived biopsy specimens of the contralateral testicle were reviewed after immunostaining for placental alkaline phosphatase (PLAP) as a marker for CIS. RESULTS: TM was diagnosed in 11 (26.8%) patients. Of them, microliths were pathologically confirmed in only 2. Prevalence of CIS was higher in patients with TM (2 of 11 patients) than in those without TM (0 of 30 patients, p = 0.017) CONCLUSION: TM was associated with testicular tumor. TM in the contralateral testis of patients with unilateral testicular tumor may increase the risk of CIS. PMID- 17419364 TI - [Efficacy on interferential low frequency therapy for elderly overactive bladder patients with urinary incontinence]. AB - AIMS OF STUDY: Examining the clinical efficacy on the interferential low frequency therapy for elderly overactive bladder (OAB) patients with urinary incontinence prospectively, for whom anticholinergics were not effective. STUDY DESIGN, MATERIALS AND METHODS: Subjects are 80 patients over 65 years with urinary incontinence, who are clinically diagnosed with OAB. For over three months, they were administered anticholinergics (20mg/day propiverine hydrochloride), but in an interview after the administration period they responded that their QOL indexes were 4 (mostly dissatisfied) or over. After the anticholinergics had been washed out for over one week, the patients were provided with interferential low frequency therapy (IF) alone for three months using the Uromaster" (made by Nihon Medix Co.). Before and after IF, the following items were examined. 1) Frequency of IF treatment required to show optimal effects subjectively, 2) Average incontinence frequency in a week, 3) PAD test for 60 minutes, 4) Frequency and voided volume in the daytime and nighttime, 5) Fluid intake volume, 6) IPSS, QOL index, 7) UFM (voiding time, voided volume, Qmax, Qave), 8) PVR (postvoid residual urine), 9) Specific gravity of urine, 10) Average hours spent outside per day, 11) Average radius of action per day, ADL scale, 12) Blood pressure and pulse in the standing position, 13) Values of clinical examination, 14) Adverse events of IF., 15) Plasma osmotic pressure, 16) BNP (Brain natriuretic peptide). During the trial, no behavioral advice, including fluid intake advice, was provided to the patients. Before starting this study, all patients signed an informed consent agreement. RESULTS: 1) After a median of eight treatments of IF, the patients showed significant improvements subjectively and this effect continued. Improvement was observed in the following items. All these values showed [The mean value before IF]-->[The mean value after IF for three months], and statistically significant differences were identified between before and after IF in all these values (p< 0.0001 except nighttime voiding frequency [p = 0.0004]). 2) Incontinence frequency; 13.3-->3.6 times/week, 3) PAD test; 17.5-3.1gr., 4) daytime voiding frequency; 8.3-*7.0 times, nighttime voiding frequency; 1.8-->1.4 times, 6) IPSS; 12.1-->6.3, QOL index; 5.2-->2.4, 7) Voided volume; 170.2-->254.2ml, Qmax; 18.1 -->25.7ml/sec, Qave; 8.9-->12.1ml/sec, 10) Average hours spent outside; 1.5-->3.0hrs., 11) Average radius of action; 400-->1,200m, ADL scale; 8.0-->3.4, 12) Blood pressure in the standing position; 132.9/ 79.7-->127.1/74.7mmHg., 15) Plasma osmotic pressure; 295.1-->297.8 mOsm/1, 16) BNP; 41.3-->19.2 pg/ ml. Other items including clinical examination and pulse did not change much, while the adverse events of IF were not recognized. INTERPRETATION OF RESULTS: The results indicate the possibility that IF has safe and better effects than anticholinergics on the elderly OAB patients with urinary incontinence. It is clear that the OAB can significantly impair the QOL and reduce the ADL. By improving the urgent incontinence, the QOL will be improved and the physical activity may increase. PMID- 17419365 TI - [Clinical importance of guideline for decontamination of endoscopes in urological field: verification of 3 processes]. AB - Clinical guideline for decontamination of endoscopes in urological field has not been provided, although endoscopic examination should be done in aseptic circumstance. MATERIAL AND METHOD: From decontamination processes, following 3 points were selected for verification: 1) Volume of residual water in flexible scope after rinsing, estimated by weight change. 2) Concentration of eluted disinfectant from flexible scope after standard rinsing procedure for gastrointestinal endoscope. 3) Observation of possible damage of telescope, caused by repeat autoclaving. RESULTS: Wet condition of flexible scope could be suspected in 60 hours in room temperature. Eluted disinfectants were detected, but in harmless level. Repeated autoclaving merely caused minor damage, which does not interfere clinical use. CONCLUSION: We obtained a couple of evidences with cautions in decontamination processes for endoscopes. Urgent requirement of standardization in this field should be discussed widely. PMID- 17419366 TI - [Association with relative nocturnal polyuria using BNP (brain natriuretic peptide) in eldely patients with nocturia]. AB - OBJECTIVE: In addition to overactive bladder (OAB) and sleep disorders (disturbance of additional sleep induction), nocturnal polyuria has been reported as an etiology of nocturia in elderly people. To investigate the influence of heart function on nocturnal polyuria in elderly people, we examined the association with nocturnal polyuria using brain natoriuretic peptide (BNP), which are useful for evaluating the prognosis of heart failure. PATIENTS AND METHODS: The patients were 128 patients (92 males, 36 females) who were treated for nocturia in Kohsei general hospital and other relative hospital between October 2002 and September 2005. We measured BNP levels at physical examination. Simultaneously, the patients were instructed to write a frequency volume chart (FVC) for 4 days. 24-hour urine volume, Daytime urine volume, nocturnal (sleep) urine volume, nocturnal polyuria index (NPi) were calculated from FVC. The association was examined. However, alphal-blockers or anticholinergic agents that had been prescribed to treat urination disorders were continuously administered. RESULTS: Overall, the mean BNP level was high, 46.3+/-39.6 pg/ml. The mean 24 hour urine volume was 1,555+/- 458 ml. The mean daytime urine volume was 935+/ 322 ml. The mean nocturnal urine volume was 624+/-251 ml. The mean nocturnal urine volume rate was high, 40.1 - 10.5%. However, there was a close association between BNP and the 24-hour urine volume (p = 0.0215), the daytime urine volume (p = 0.0004), the NPi (p = 0.0003). The daytime urine volume decreased with the BNP level. The NPi increased with the BNP level. Patients were divided into 2 groups, a group with a BNP level less than 50 pg/ml and a group with a BNP level of 50 pg/ml or more. In the group with a BNP level less than 50 pg/ml, the nocturnal urine volume rate was 38.14+/-10.07%. In the group with a BNP level of 50 pg/ml or more, the rate was significantly higher (43.97+/-10.48%, p<0.0029). CONCLUSIONS: These results suggest that many elderly patients latently have mild heart failure, and that relative nocturnal polyuria reduces cardiac load. Therefore, in patients with a high BNP level, administration of antidiuretic hormone to decrease nocturnal urine volume is risky. Administration of diuretics during the afternoon or evening may be safer. PMID- 17419367 TI - [Preoperative parameters, including percent of positive biopsy cores, in predicting pathological findings after radical prostatectomy]. AB - OBJECTIVES: We investigated whether preoperative parameters predict pathological stage at radical prostatectomy for patients with clinically localized prostatic cancer. MATERIALS AND METHODS: We studied a total of 160 men with clinically localized prostatic cancer (less than or equal to clinical T2) who underwent radical rertropubic prostatectomy at Wakayama Medical University. Clinical Ts patients are not included in this study. Preoperative parameters include patient age, Body Mass Index, preoperative serum PSA value, biopsy Gleason score, clinical stage, the percent of positive biopsy cores (%PosBx) and the percent of positive biopsy cores on the dominant side (%DomPosBx). Univariate and multivariate analysis were performed to examine the prognostic significance of these preoperative parameters. Significant independent factors were combined to create a table to predict pathologically organ confined disease. RESULTS: Univariate analysis showed preoperative serum PSA value (p< 0.001), biopsy Gleason score (p =0.001), clinical stage (p = 0.026), %PosBx (p= 0.002) and %DomPosBx (p=0.003) were significantly related to the pathological stage. On multivariate analysis, serum PSA value (p< 0.01), biopsy Gleason score (p<0.05) and %DomPosBx (p<0.05) were significant independent predictors of pathological stage. CONCLUSION: We provide two model combinations using preoperative clinical factors, one is a combination of serum PSA and biopsy Gleason score and the other is a combination of serum PSA and %DomPosBx, which define a new preoperative model for predicting pathological organ confined prostatic cancer. These combinations are useful and provide important information for urologists to determine the appropriate treatment strategy for clinically localized prostatic cancer. PMID- 17419368 TI - [A case of small cell carcinoma in a diverticulum of the bladder]. AB - We report a case of small cell carcinoma in a diverticulum of the bladder. A 64 year-old Japanese man visited our hospital with the chief complaint of urinary retention. Cystoscopy revealed a papillary tumor arising in a diverticulum on the right lateral wall of the bladder. Partial cystectomy was performed under the diagnosis of bladder cancer in the diverticulum. The histopathological diagnosis was a combination of small cell carcinoma and urothelial carcinoma, of which the former was dominant. It is often difficult to detect tumors in the bladder diverticula and its early penetration is not rare because of the thin wall. Therefore, the prognosis of the bladder cancer in the diverticulum is believed to be generally poor. On the other hand, small cell carcinoma originating in the urinary bladder is also characterized by its extreme rareness and poor prognosis. To our knowledge, this is the second case report of the small cell carcinoma arising in a diverticulum of the urinary bladder in the Japanese literature. PMID- 17419369 TI - [A case report of sarcomatoid carcinoma of the urinary bladder]. AB - A 49-year-old female visited to our department with complaints of gross hematuria and micturition pain. Cystoscopic examination revealed a 5-cm diameter solid broad-based tumor located at the front wall of the bladder. Histologically, the tumor was composed of grade 2 urothelial carcinoma (UC) element and sarcomatous spindle cell element. Immunohistochemical examination demonstrated that both UC and spindle cells were positive for cytokeratin. In addition, spindle cells did not stain for S-100 protein or smooth muscle actin. We diagnosed the tumor as sarcomatoid carcinoma and performed total cystectomy and ileal conduit without chemotherapy and radiation. The patient has remained without any evidence of recurrence for 14 months after operation. Sarcomatoid carcinoma of the bladder has aggressive malignant potential and poor prognosis. An appropriate adjuvant therapy for sarcomatoid carcinoma with metastasis has not been established. Total cystectomy is recommended as soon as possible after pathological diagnosis in the same manner as conventional high grade UC. PMID- 17419370 TI - [Radical prostatectomy for prostate carcinoma with ectopic ureter ; a case report]. AB - A 71-year old male visited our hospital with a chief complaint of pollakisuria. The needle biopsies of the prostate were performed with PSA 8.0 ng/ml, and he was diagnosed as moderately differentiated adenocarcinoma. Imaging techniques revealed a right complete duplicated upper urinary system with an ectopic ureter draining to the prostatic urethra. He received radical prostatectomy with concomitant anastomosis of ureter to ureter. There is no evidence of hydronephrosis or tumor recurrence 11 months after operation. This is, to our knowledge, the second case report describing the association of radical prostatectomy and ectopic ureter. PMID- 17419371 TI - [COPD as a systemic disease]. PMID- 17419372 TI - [Historical review on changes in the concept of COPD]. AB - A brief review of changes in the concept of COPD was made based on records of mile stone articles and prestigious international lung conferences. A major innovation in the definition of COPD was presented by the 'GOLD WORKSHOP REPORT' published in 2001. It introduced a concept of inflammatory response of the lung to toxic gas/particles as the most significant pathogenesis of the disease, eliminating the definition of COPD being a syndrome of 'emphysema plus chronic bronchitis'. Both CIBA Guest Symposium and Aspen Emphysema Conference contributed in the evolution of COPD concept. Also discussed in brief are the impacts of 'Small Airway Disease Theory' and British bronchitis hypothesis. The GOLD revision 2006 may be highly appreciated among practicing clinicians world wide with key features; 'COPD is preventable and treatable accompanied by significant systemic effects'. PMID- 17419373 TI - [Guideline for COPD management and prevention]. AB - Chronic obstructive pulmonary disease (COPD) is recognized as very important disease in high prevalence and leading cause of death in the world. The new 2006 GOLD report suggests that COPD is a preventable and treatable, and should be treated, depending on the severity of the disease and the clinical status of the patients. The report also indicates that primary care practitioners treat the vast majority of COPD patients and may be actively involved in public health campaigns and in bringing messages about exposure to risk factors to both patients and public. PMID- 17419374 TI - [Epidemiology of COPD in Japan: NICE study (Nippon COPD Epidemiology Study)]. AB - Although chronic obstructive pulmonary disease (COPD) has high prevalence and mortality, there are underestimated and many COPD patients still remain not diagnosed worldwide including Japan. In spite of high prevalence in Western countries, it had been reported that COPD patients were estimated about 200,000 in Japan. Here, the prevalence of COPD in Japan is discussed according to the NICE Study. PMID- 17419375 TI - [Effects of smoking on the pathogenesis of COPD]. AB - Cigarette smoking is the major risk factor for COPD. However, it is likely that there are important interactions between environmental factors, such as cigarette smoking, and a genetic predisposition to COPD. Single cigarette smoking causes acute reversible airway inflammation in everyone, and only 15 % of chronic smokers may develop chronic irreversible airway inflammation, resulting in the development of COPD. Fifteen percent reason has been undefined. Multiple factors may involve the pathogenesis of COPD. Cigarette smoke may upregulate the pathways linked to lung destruction and airway inflammation, such as protease and oxidative stress, and may downregulate the pathways related to the defensive mechanism, such as antiprotease and antioxidant. Other than protease-antiprotease and oxidant-antioxidant mechanisms, multiple exposures to cigarette smoke move cells into an irreversible state of senescence, or the inability to repair lung injury. Chronic exposure to smoking may affect the VEGF signaling, resulting in apoptosis of lung cell, which may partly contribute to the pathogenesis of COPD. PMID- 17419376 TI - [Progress in the genetics of chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is a multifactorial disease caused by the interaction of genetic susceptibility and environmental factors. Although cigarette smoking is the main environmental risk factor for developing COPD, only about 15 % of smokers develop clinically significant disease. Genetic researches in this field mainly focused on variants of genes involved in protease anti protease systems, defense against oxidative stress and inflammation. In addition to the candidate gene studies, genome wide approach including genome wide association studies will be extensively done in the near future. PMID- 17419377 TI - [Childhood respiratory infection in the pathogenesis of COPD]. AB - A respiratory infection in childhood has been implicated in one of the risk factors for COPD. The hypothesis that latent adenoviral infection is involved in the pathogenesis of COPD have been studied for 15 years. A DNA virus, adenovirus(Ad), early gene 1A (ElA gene) was identified in lung tissue of COPD. The E1A gene expression enhances the soluble intercellular adhesion molecule (ICAM) 1 expression and the recruitment of inflammatory cells into airways of COPD lungs. In vitro experiments, E1A gene transfected cells exhibited the enhanced inflammatory response to tobacco smoke. The excess production of ICAM-1, IL-8, and TGF- beta by lung epithelial cells transfected with the Ad E1A gene. Respiratory infections in childhood including Ad infection may be an important risk factor for the pathogenesis of COPD. PMID- 17419378 TI - [Role of mediators in the onset of COPD]. AB - Chronic obstructive pulmonary disease (COPD) is a chronic airway disorder characterized by obstructive airflow limitation which is not completely reversible with treatment. Inflammatory changes in the peripheral airways, especially those with the diameter less than 2 mm(so called small airway disease) have been speculated to be initial steps of COPD. However, it remains unclear which cytokines/chemokines play an essential role in the pathogenesis of these lesions. Studies with bronchoalveolar lavage demonstrated an increase in neutrophil numbers and the neutrophil chemoattractant interleukin-8. Recent studies demonstrated that T lymphocytes, especially CD8 (+) cells are increased and express a variety of cytokines and chemokines, which are involved in cell infiltration and activation. Studies with gene-engineered animals and anti cytokine treatment will facilitate better understanding the role of cytokines/chemokines, and eventual novel therapy. PMID- 17419379 TI - [Role of alveolar cell apoptosis in COPD]. AB - Chronic obstructive pulmonary disease (COPD) is a cigarette smoking-related disorder characterized by chronic inflammation of the airways and progressive destruction of lung parenchyma, leading to airway remodeling and pulmonary emphysema. Several mechanisms have been proposed to be involved in the pathogenesis of pulmonary emphysema, including an imbalance of proteases and antiproteases, and oxidative stress. In addition to these mechanisms, recent studies suggest another mechanism involved in the development of pulmonary emphysema: apoptosis of alveolar wall cells. There is an increase in apoptotic alveolar epithelial and endothelial cells in the lungs of patients with emphysema, and since this is not counterbalanced by an increase in proliferation of alveolar wall cells as they become senescent, the net result is loss of alveolar unit leading to the development of emphysema. Studies with animal models suggest that caspases, vascular endothelial growth factor (VEGF) deficiency, oxidants, ceramide, CD8+ T-lymphocytes, elastases, and interferon-gamma may be responsible for the induction of apoptosis of alveolar wall cells. Furthermore, defective clearance of apoptotic cells in the lungs of patients with emphysema may promote inflammation. In this review, recent data on the role of apoptosis in emphysema from both animal models as well as from studies on human subjects will be discussed. PMID- 17419380 TI - [Oxidative stress in pathogenesis of COPD]. AB - Cigarette smoke and aging are major risk factors of chronic obstructive pulmonary disease(COPD). It remains unsolved how long -term smoking with age affects the molecular responses in the lung. Respiratory tract is the major interface to the environment and is rich in glutathione, which protects lung from oxidative stress. We performed bronchoalveolar lavage for nonsmokers and smokers of various ages, who were further categorized according to the presence of emphysema on high resolution computed tomography. We thus evaluated glutathione antioxidant system in BAL fluid. Characterization of older smokers with long-term smoking histories, contrasted with young recent smokers, may in part explain the predisposition of the lungs to destructive lung diseases. On the other hands, oxidative stress results from an imbalance in aerobic metabolism and poses a serious threat to cellular apoptosis, leading to emphysematous lung destruction. The therapeutic interference with targeted up-regulation of protective mechanisms might be critical for the success of future COPD therapies. PMID- 17419381 TI - [Characteristics of pathophysiology in COPD]. AB - Chronic obstructive pulmonary disease (COPD) is characterized by airflow limitation caused by emphysema and/or airway narrowing. The major site of airflow limitation in COPD is thought to be small airways (inner diameters<2 mm). In patients with COPD, FEV1 (%predicted) is highly correlated with airway luminal area and also with wall thickening and the correlation coefficients improve as airway size decreases. In patient with COPD, maximal flow(V max) at lower lung volumes is remarkably lowered. As the disease progresses, air trapping occurs even during resting ventilation, thus causing increased functional residual capacity (FRC). Reduced inspiratory capacity as a result of increased FRC is believed to be highly correlated with breathlessness in COPD. PMID- 17419382 TI - [Airway inflammation in COPD]. AB - COPD is characterized by chronic and persistent inflammation in the small airways and lung parenchyma, which lead to airway remodeling and parenchymal destruction. This inflammation is mediated by the increased expression of multiple inflammatory genes. For the initiation of the transcription of inflammatory genes, the acetylation of core histones is necessary to open up the chromatin structure, that allow transcription factors and RNA polymerase to bind to DNA. Conversely, deacetylation of these histones switches off for the gene transcription. In the COPD lung, the reduced expression of histone deacetylases (HDACs), particularly HDAC-2 is thought to be a key mechanism for the persistence of airway inflammation and corticosteroid resistance. PMID- 17419383 TI - [CT imaging of COPD--importance of phenotyping (emphysema dominant and airway disorder dominant)]. AB - We have been engaged in the analysis of the distribution of low attenuation area (LAA) representing emphysema, and the measurement of airway dimensions using CT images from the view point of phenotyping of COPD. (1) L AA% (area ratio of LAA to all lung area) did not correlate with reversibility to bronchodilators, whereas WA% (area ratio of bronchial wall to cross section of bronchus) positively correlated with reversibility. Thus, bronchodilators may be effective in proportional to the extent of airway disorders. (2) The incidence of Gc*1F(+) was significantly higher in patients with severe emphysema. These results indicate that the patients with Gc*1F(+) have rapid development of emphysema. (3) Polymorphism of MMP-9 (C -1562T) was associated with upper lung dominant emphysema in patients with COPD. (4) CT measurements of emphysema were greater in the patients with anorexia nervosa than in historical control subjects (p<0.001). Furthermore, there were significant correlations between the body mass index and the CT measures of emphysema. These data demonstrate that emphysema-like changes are present in the lungs of patients who are chronically malnourished. PMID- 17419384 TI - [COPD: criteria for diagnosis and classification of disease severity]. AB - In the guideline proposed by WHO (GOLD) or by the Japanese Respiratory Society (RS), the diagnosis of COPD is simply made when FEV1/FVC (FEV %) of a given subject after inhalation of short-acting bronchodilator is below 70%. On the other hand, on grounds of measured FEV1/predicted FEV (%FEV1), the disease severity of COPD is categorized into four stages including I (mild), II (moderate), III (severe), and IV(very severe). In addition, very special category defined as stage zero (at-risk group) has been proposed in both guidelines. However, the above-mentioned criteria have many impediments as they are too simple for diagnosing and classifying COPD with a complicated pathophysiology. In this paragraph, the attempt is made not only to comment on the diagnosis criteria and classification of disease severity of COPD provided in GOLD and JRS guidelines but also to investigate the details of the impediments existing in both guidelines. PMID- 17419385 TI - [Spirometry and other pulmonary function tests for the screening and evaluation of patients with chronic obstructive pulmonary disease (COPD)]. AB - Chronic obstructive pulmonary disease (COPD) is a disease state characterized by airflow limitation that is not fully reversible. The existence of airflow limitation can be determined by spirometry that measures the forced expiratory volume in one second (FEV1) and its ratio (FEV1/FVC) to the forced vital capacity (FVC). Airflow limitation is defined as FEV1 /FVC<70% after the inhalation of short-acting bronchodilator. In addition, FEV1% predicted, a ratio of FEV1 to the predicted value determined by age, gender and height, is utilized to define the severity degree of COPD. Some of the other pulmonary function tests are useful for understanding the pathophysiology of COPD. These tests include the diffusing capacity measurement of carbon monoxide per liter of alveolar volume (DLco/VA), measurement of lung volume using the nitrogen washout technique and whole body plethysmography, and measurement of lung compliance. PMID- 17419386 TI - [Radiological diagnosis --diagnosis and evaluation by chest X ray, chest CT and chest MRI]. AB - A plain chest X-ray is not useful for the diagnosis of early stage of COPD, but severe COPD has some characteristic radiographic signs. Because of absence of vasculature, radiolucency of lung fields is recognized. Depression and flattening of the diaphragm, tear -drop heart and increase in the retrosternal airspace are detected as signs of lung hyperinflation. High-resolution CT(HRCT) scanning is more sensitive to diagnose of COPD. COPD can be detected as low attenuation area. CT is also useful in classifying patterns of emphysema(centriacinar, panacinar and distal acinar type). Although the assessment of COPD is still limited by MRI, progress of MRI techniques in pulmonary ventilation, such as hyperpolarized noble gas MRI and oxygen enhanced MRI, enabled evaluation of COPD. PMID- 17419387 TI - [Diseases to differentiate from COPD, with emphasis on bronchial asthma]. AB - Differential diagnosis of chronic obstructive pulmonary disease (COPD) from asthma is not a difficult task for many clinicians. Patients with COPD have a history of heavy smoking and show a slowly progressive dyspnea on exertion and there is little variability in symptoms, and they show a poor response to bronchodilators and corticosteroids. Asthma usually begins in early childhood with atopy, shows episodic dyspnea with wheezing, especially during night and early morning. Some patients, however, show adult onset, irreversible airflow limitation, and neutrophilic airway inflammation. The airway remodeling in asthma may be the cause of confusing pathophysiology. Other diseases showing airway hyperresponsiveness, such as allergic bronchopulmonary aspergillosis, Churg Strauss syndrome, and left heart failure presenting cardiac asthma, may sometimes show similar clinical pictures to COPD. Chronic airway diseases are also possible candidates for differential diagnosis of COPD. Bronchiectasis, sinobronchial syndrome, diffuse panbronchiolitis, obliterative bronchiolitis, and other chronic airway diseases should be considered. Some interstitial lung diseases, such as smoking-related interstitial lung diseases and lymphangioleiomyomatosis, often show obstructive ventilatory impairment, and therefore should be considered in differential diagnosis of COPD. PMID- 17419388 TI - [A stepwise increase in pharmacotherapy depending on the severity of COPD]. AB - The overall approach to managing stable COPD should be characterized by a stepwise increase in treatment, depending on the severity of the disease in accordance with JRS COPD guideline. None of the existing medications has been shown to modify the long term decline in lung function that is the hallmark of this disease. Therefore, pharmacothepary for COPD is used to decrease symptoms and/or complications. Smoking cessation and influenza vaccines are the base of pharmacotherapy. And the principal short and long acting bronchodilator treatments are anticholinergics, beta2-agonists, methylxanthines, and a combination of one or more of these drugs. And regular treatment with inhaled glucocorticosteroids should only be prescribed for symptomatic COPD patients with a documented spirometric response to glucocorticosteroids or for those with stage III and repeated exacerbations. PMID- 17419389 TI - [New development of pharmacotherapy--physiological basis and future possibility]. AB - In COPD management, currently available treatments improve the quality of life (QOL) of the patients. According to the international and Japanese guidelines, long-acting bronchodilators are central to the management of COPD. Recently developed long-acting anti -cholinergic agent and beta2-agonists show remarkable effects on COPD subjects compared with short-acting agents. The physiological explanation of the efficacy of long-acting bronchodilators seems to be due to the pharmacological volume reduction via airway dilation. In future, we need COPD specific anti-inflammatory agents, including histone deacetylase (HDAC) activator. PMID- 17419390 TI - [Clinical efficacy of inhaled corticosteroids in COPD]. AB - The use of inhaled corticosteroids in asthma is regarded as first-line therapy. But the efficacy of inhaled corticosteroids in the treatment of chronic obstructive pulmonary disease (COPD) remains controversial. However data from some large studies provide the evidence that regular treatment with inhaled glucocorticosteroids is appropriate for symptomatic COPD patients with an FEVi<50% predicted and repeated exacerbations. In these studies glucocorticosteroids combined with a long-acting beta-agonist was more effective than the individual components. However, additional studies are needed to clarify the effects of inhaled corticosteroids on mortality and to define their long-term adverse effects. PMID- 17419391 TI - [Pulmonary rehabilitation--exercise training and nutritional support]. AB - Exercise training, widely regarded as the cornerstone of pulmonary rehabilitation, is the best available means of improving exercise capacity, exertional dyspnea, and activities of daily living in COPD. However, optimal intensity of training, long-term benefits and more efficient ways for severe patients with COPD remain uncertain. Adjunctive strategies, such as neuromuscular electrical stimulation (NMES) and noninvasive positive pressure ventilation (NPPV), must be developed. Although nutritional status is an important determinant of symptoms, disability, and prognosis in COPD, present evidence suggests that nutritional supplementation alone may not be sufficient strategy. Nutritional support to be accompanied by exercise training and to attenuate systemic inflammation should be established. In addition, administration of orexigenic factor(e.g. ghrelin) may be new strategy for COPD patients with nutritional depletion. PMID- 17419392 TI - [Home oxygen therapy]. AB - Oxygen therapy is one of the principal non-pharmacologic treatments for severe chronic obstructive pulmonary disease (COPD) patients. Home oxygen therapy(HOT), or long-term oxygen therapy(LTOT) for 15 hours or more per day, can improve the survival rate of severe COPD patients with beneficial effects on hemodynamic state, hematological characteristic, exercise capacity, lung mechanics, and mental state. Oxygen therapy is indicated in cases of severe chronic respiratory failure with PaO2 of 55 Torr or less, or in cases with PaO2 of 60 Torr or less in whom there is remarkable hypoxia during sleep or during exercise. The induction of oxygen therapy needs evaluations of oxygen desaturation during exercise and sleep as well as hypoxia at rest. It also required to consider CO2 narcosis. PMID- 17419393 TI - [Ventilatory assist therapy]. AB - NPPV has been shown to improve arterial blood gas values, prevent symptoms resulting from alveolar hypoventilation, and decrease hospitalization in patients with chronic respiratory failure with COPD. Regarding acute exacerbation of COPD, the utility of NPPV has been established so NPPV will be regarded first-line therapy as ventilatory assist therapy. On the other hand, with respect to chronic stable COPD, the utility of NPPV isn' t established yet. However NPPV cases have been increasing rapidly in these years, as new home respiratory care technique. From now on, we have to evaluate the objective utility of NPPV from the standpoint of evidence. PMID- 17419394 TI - [The programs for smoking cessation]. AB - Smoking is a major risk factor of the chronic obstructive pulmonary disease (COPD) and increases aging changes of pulmonary function. Also the passive smoking for children increases the prevalence of several pulmonary diseases. It is very important to stop smoking for the prevention and treatment of COPD and other pulmonary diseases. In the results of our study, almost of smokers don' t know the influences of smoking exactly, and a half of them want to stop smoking if they have a chance. The useful programs are developed and outpatient clinics for smoking cessation are increasing in Japan. I would like to show some methods and problems. PMID- 17419395 TI - [Surgical treatment for pulmonary emphysema--LVRS/lung transplantation]. AB - Lung volume reduction surgery (LVRS) for severe pulmonary emphysema was started more than 10 years ago. However, its indication and durability of effects had not been clear. The National Emphysema Treatment Trial (NETT) defined patients at high risk of death after LVRS and subgroups of patients with severe emphysema in whom LVRS improved survival and function. Those effects are durable, when LVRS is applied to an appropriate subgroup like upper lobe predominant emphysema patients with low exercise capacity. However, only one-fourth of LVRS candidates are belonging to such a preferable subgroup. Although most of the patients with severe pulmonary emphysema are over 60 years old in Japan, candidates for lung transplantation are limited under 60. Under such a condition, only two patients with severe pulmonary emphysema have undergone lung transplantation in Japan. PMID- 17419396 TI - [Treatment and prevention of COPD exacerbation]. AB - Airway inflammation, mucosal edema, epithelial hyperpermeability, mucus secretion and airway smooth muscle contraction induced by airway bacterial, virus infection and exposure to air pollution may be associated with COPD exacerbation. Severity of COPD exacerbation is estimated by blood gas analysis, serum CRP values and the chest radiograph. Patients with COPD exacerbations are recommended to be treated with additional inhalations of beta-2 agonists and anti -cholinergic agents, systemic administered glucocorticosteroids, oxygen inhalation, and, in cases with purulent sputum, antibiotics. Glucocorticosteroids, beta-2 agonists and anti cholinergic agents reduce the frequency of COPD exacerbation. We reported the inhibitory effects of glucocorticosteroids on rhinovirus infection, the major cause of common colds, and the inhibitory effects of L-carbocisteine and erythromycin on COPD exacerbations and rhinovirus infection. PMID- 17419397 TI - [Cell based therapy for COPD]. AB - To develop a new cell based therapy for chronic obstructive pulmonary disease (COPD), we need to understand 1) the role of tissue-specific and bone marrow derived stem cells, 2) extracellular matrix, and 3) growth factors. Recently, bronchioalveolar stem cells were identified in murine distal lungs. Impairment of these stem cells may cause improper lung repair after inflammation, resulting in pulmonary emphysema. Bone marrow-derived cells are necessary to repair injured lungs. However, the long term role of these cells is not understood yet. Although we need more careful analysis and additional experiments, growth factors, such as hepatocyte growth factor, are good candidates for the new cell based therapy for COPD. Lung was believed as a non-regenerative organ. Based on these recent reports about lung regeneration and stem cells, however, new strategies to treat COPD and a new point of view to understand the pathophysiology of COPD are rising. PMID- 17419398 TI - [Team medicine for COPD]. AB - From the standpoint of holistic respiratory care for patients with COPD, team medicine by various kinds of occupations are required. Patient education and guidance are very important at the introduction of HOT and HMV. Moreover, for smooth and safe home respiratory care, local health care network is essential. We secure nursing visit and local physicians as much as possible in order to confirm adherence after starting home care and for early discovery of problems. Delicate care through a local health care network is important. However there is insufficiency regarding the establishment of local health care network, so it should be improved immediately. PMID- 17419399 TI - [Wearable chemical sensors for non-invasive monitoring]. AB - Non-invasive and non-restrictive bioinstrumentation is strongly requested in the field of medical and health care technologies, being under rapid advance in informatization. Applying "Soft-MEMS" techniques onto functional polymers, flexible chemical sensors were developed. This sort of sensors is one of the promising candidates that realize the requirements. Wearable oxygen sensor was first introduced and applied to transcutaneous monitoring of partial pressure of arterial oxygen. Then, wearable glucose sensors, which were constructed by immobilizing glucose oxidase onto the sensing region of a wearable oxygen sensor were reported. Considering the progress towards contact lens type sensor in mind, optical-transparent was developed. On the other hand, biocompatible polymer was introduced. Both sensors were enough sensitive to measure glucose concentration in tear fluids (0.14 mmol/l). In this paper, recent researches on wearable oxygen sensors, wearable glucose sensors and their applications are reported. PMID- 17419400 TI - [Cerebrovascular disease and diabetes mellitus]. AB - In Japan, individuals with diabetes mellitus have been increasing rapidly. A population -based study, conducted in a Japanese community, Hisayama, in 1988, showed that among subjects aged 40 to 79 years, the prevalence of glucose intolerance: namely impaired fasting glycemia, impaired glucose tolerance, and diabetes, exceeded 40% in men and 30% in women. Previous cohort studies in the world demonstrated that the multivariate -adjusted relative risks of stroke (cerebral infarction) for diabetes were significantly high and ranged from 2 to 5. Diabetes induces various types of cerebral infarction and significantly increases the recurrence and mortality rates after first episode of stroke. Hyperglycemia and insulin resistance observed in diabetes cause arteriosclerosis through various mechanisms, resulting in the development of stroke. PMID- 17419401 TI - [Concepts and theory of health impact assessment]. AB - Health effects are often overlooked in the planning of policies, programmes or projects, which has led to international and national pressure for evaluation of potential influence. For this reason, Health Impact Assessment (HIA) has been emphasized by many national governments and international organizations such as the European Union and WHO. HIA is a helpful decision-making tool with methodology that was defined as "a combination of procedures, methods, and tools by which a policy, a program or a project may be judged as to its potential effects on health of a population and the distribution of effects within the population" in the WHO Gothenburg paper. In recent decades, many HIAs have been implemented for proposals including building new airports, dams, employment strategy, and housing policy. However, there is very little information on HIA in Japan, even among public health professionals and policy makers. In this review, we introduce basic concepts and theory, and discuss how to improve HIA activities in Japan. PMID- 17419402 TI - [A study of factors related to activities of daily living (ADL) of the elderly receiving in-home service longitudinal study using functional independence measures]. AB - OBJECTIVE: We conducted a longitudinal study using Functional Independence Measures to clarify factor related to independence of activities of daily living of elderly receiving in-home service under the long-term care insurance system METHODS: Fifty-four elderly users of the in-home service of Ibaraki Prefecture assented to participate in this study and were analyzed. A researcher conducted survey at the baseline and after follow-up by visiting the elderly at each home. The evaluation standards used here were the Japanese version of Functional Independence Measure (FIM), Mini-Mental State Examination (MMSE), and Geriatric Depression Scale (GDS-15). RESULTS: The FIM score (mean+/-SD) was decreased 83.6+/-36.4 to 81.7+/-37.4 during the 112+/-22.2 day follow up period. Thirty nine elderly demonstrated improvement or no change in FIM and 15 had declining scores. To clarify independent factors related to FIM change, we conducted a step wise multifactor logistic regression analysis, and the results suggested importance for "in-home service availability" and "home care period less than one year". CONCLUSION: Our study suggested that it is important for maintenance or improvement of ADL in home care elderly to provide sufficient home .care services from the beginning under the long-term care insurance system. PMID- 17419403 TI - [A measure of the motives underlying snack selection among Japanese junior high school students: the Snack Choice Questionnaire (SCQ)]. AB - PURPOSE: To develop a measure of the motives underlying snack selection by Japanese junior high school students and to examine the characteristics of each motivating factor. METHODS: Self-reported questionnaires were distributed in a cross-sectional study of 1,936 students in public junior high schools in Tokyo, Japan. The respondents answered the Snack Choice Questionnaire (SCQ) and the Dutch Eating Behavior Questionnaire (DEBQ), which assess overeating, snacking behavior, the food environment, lifestyle, and demographics. RESULTS: Twenty-two items of the SCQ were factor-analyzed using varimax rotation. Three factors were extracted and labeled "fashion and sales promotion," "convenience and taste," "health and weight control." All factors demonstrated a satisfactory Cronbach's alpha coefficient of over 0.80, and scores for both "fashion and sales promotion" (r= 0.349, P<0.001) and "convenience and taste" (r= 0.418, P<0.001) showed positive correlations with DEBQ scores. On the other hand, scores for "health and weight control" (r= 0.014, ns) were not statistically significant. Furthermore, the students who reported buying or eating snack foods frequently had high scores for "fashion and sales promotion" and "convenience and taste" but not for "health and weight control." The factor "fashion and sales promotion" was related to more TV viewing (beta = 0.060, P< 0.05), while the factor "convenience and taste" was related to the availability of convenience stores near a student's home (beta=0.109, P<0.001). CONCLUSIONS: The results of this first study of the motives underlying snack food selection in junior high-schools in Japan suggest a need for comprehensive nutrition education, along with a focus on media literacy and consumer education. PMID- 17419404 TI - [Risk factors for reduced visual acuity in female junior high school students: findings of a cross-sectional study]. AB - PURPOSE: The purpose was to explore the risk factors for reduced visual acuity in female junior high school students that are relevant to their living habits/environment. METHODS: A cross-sectional survey was conducted to collect data on the living habits/environment and visual acuity of female junior high school students. All the variables measured in the study were categorized into multiple, mostly two, categories, since distributions of data that are obtained at school medical examinations are often right-skewed with possible outliers; in particular, application of accurate measurement of visual acuity such as the method of refractive power was not feasible. Multiple logistic regression was employed in the analysis of data, incorporating the graphical modelling technique. Akaike Information Criteria (AIC) was used to select the best model fit for the data. RESULTS: The odds ratio for students watching TV from less than a 2 m distance relative to more than 2 m was 2.08 (95%CI 1.23-3.50) and statistically significant, after adjustment for the study time at home/cram school, reading time, sleeping time, and use of glasses/contact lenses by parents/siblings. On the other hand, after the same adjustment, the odds ratio for students watching the TV for 2 h or more relative to those watching the TV for less than 2 h was close to unity and not even selected in the final model. The results also indicated that study time at home/cram school, reading time, and use of glasses/contact lenses by parents/siblings are significantly associated with reduced visual acuity. The risks of combinations of these variables were evaluated by means of the odds ratios. CONCLUSION: We found several risk factors that are responsible for reduced visual acuity in female junior high school students. It is considered that the distance of TV viewing is the most important and yet relatively amenable to intervention for improvement to avoid reduction in visual acuity. Multivariate evaluation of the risks for visual acuity decrease was possible by means of odds ratios. Concrete suggestions can thus be made to female junior high school students to prevent decrease of visual acuity by changing the living habits/environment. PMID- 17419405 TI - [A nationwide survey for implementation of health Japan 21 dental health promotion programs in local municipalities]. AB - PURPOSE: The purpose of this study was to access the present situation regarding implementation of dental health promotion activities of Health Japan 21 in municipalities. METHODS: Of 1,446 local municipalities that had established local plans for the Health Japan 21 project in 2003, 953 were eligible because these had not been consolidated nor were plan had for consolidation by March 2006. A questionnaire form was mailed to the municipalities in July 2005 with items for assessment of the present status, setting goals for the project, and expectations regarding accomplishment of the local project for Health Japan 21. RESULTS: A total of 788 local municipalities responded by September 2005 (response rate 82.7%). Of them, 638 had already finished development of the local plan. As to the present status, 94% covered dental caries screening for 3 year old children. In contrast, only 25% included screening for adult paradentitis. Two thirds of municipalities set goals for dental health of 3 year old children. However, only had set any goal for the participation rate with regard to periodic dental examinations for school children (6.8%) or the prevalence of advanced paradentitis (16.3%). Proportions of municipalities that expected difficulty in accomplishing the goals were 10.1% for dental caries prevention for 3 year old children, and 30.1% for facilitation of use of dental and periodontal care equipment. CONCLUSIONS: This nationwide survey revealed dental health programs in municipalities to be relatively well implemented for dental caries prevention, but with low rates for adoption and implementation of the periodontal disease prevention program of Health Japan 21. PMID- 17419406 TI - [Smoking status among Japanese pregnant women]. PMID- 17419407 TI - [Efficacy of lipid reduction therapy in patients with coronary disease]. PMID- 17419408 TI - [Latest chemotherapy of cancers of the digestive system]. PMID- 17419409 TI - [Newest chemotherapy of rheumatoid arthritis]. PMID- 17419410 TI - [Diagnosis and treatment of diabetic nephropathy--the current status of management and its problem]. PMID- 17419411 TI - [Treatment of diabetes mellitus based on EBM]. PMID- 17419412 TI - [Neurological internal medicine in the EBN era--recent progress in treatment of adrenoleukodystrophy]. PMID- 17419414 TI - [Chemotherapy of chronic obstructive pulmonary disease]. PMID- 17419413 TI - [Guideline for the care of hematologic diseases based on EBM]. PMID- 17419415 TI - [Diagnosis and treatment of diseases of the small intestine]. PMID- 17419416 TI - [Early diagnosis and non-surgical treatment of cancers of the stomach and large intestine]. PMID- 17419417 TI - [Progress in chemotherapy of type 2 diabetes mellitus]. PMID- 17419418 TI - [Diagnosis and treatment of hyperuricemia and gout]. PMID- 17419419 TI - [Clinical aspect of peripheral nerve disorders]. PMID- 17419420 TI - [Progress in the studies of Alzheimer's disease]. PMID- 17419421 TI - [Progress in diagnosis and treatment of allergic diseases]. PMID- 17419423 TI - [Progress in treatment of osteoporosis]. PMID- 17419422 TI - [Important organ lesions associated with collagen diseases]. PMID- 17419424 TI - [Biological information monitoring and treatment of cardiac diseases]. PMID- 17419425 TI - [Information on cardiopulmonary resuscitation that is necessary for internists- current information]. PMID- 17419426 TI - [Diagnosis and treatment of drug-induced pulmonary disorders]. PMID- 17419427 TI - [Diagnosis and treatment of idiopathic pulmonary fibrosis]. PMID- 17419428 TI - [Diagnosis and treatment of malignant lymphoma]. PMID- 17419430 TI - [Factors responsible for progression and exacerbation of glomerulonephritis and their control]. PMID- 17419429 TI - [Treatment of leukemia]. PMID- 17419431 TI - [Physiopathology and treatment of renal failure]. PMID- 17419432 TI - [Current topics concerning emerging and re-emerging bacterial infections]. PMID- 17419433 TI - [Clinicopathological differences between acute and chronic eosinophilic pneumonia]. AB - Considerable confusion exists regarding the proper classification of idiopathic eosinophilic pneumonia (IEP). In addition, there are no reports that reveal clinicopathological differences between the various eosinophilic pneumonias. A problem persists in describing what the essential histological differences are between the different types of IEP. In this context, we examined the histological findings of acute eosinophilic pneumonia (AEP) and chronic eosinophilic pneumonia (CEP) and contrasted them with the clinical features and radiological findings. Radiologically, ground glass opacity and interlobular septal thickening were characteristic of the AEP cases studied, while air space consolidation was seen in all CEP cases. Histologically, interstitial edema and fibrin deposition were prominent in the AEP cases. Type II cells were detached from the alveolar walls, though the basal lamina was predominantly intact. In CEP, in addition to cellular infiltration, there was prominent intraluminal fibrosis. Disruption of the basal lamina was observed and nests of intraluminal fibrosis were directly adjacent and connected to the alveolar walls. From these findings, we conclude that the histological differences between AEP and CEP are the severity of basal lamina damage, the amount of subsequent intraluminal fibrosis, and the severity of interstitial edema. Especially in AEP, interstitial edema is an essential histological finding and this finding explains the acute onset, and the radiographic findings, as well as the rapid and complete improvement noted in such cases. PMID- 17419434 TI - [Testing for Mycoplasma pneumonia using the ImmunoCard Mycoplasma rapid test]. AB - We evaluated the effectiveness of ImmunoCard Mycoplasma rapid tests in all patients admitted with community-acquired pneumonia (CAP) between January, 2004 and December, 2005. ImmunoCard Mycoplasma rapid tests were performed on the 1st day of admission and we analyzed the frequency of positive cases among CAP cases according to month and age. A total of 82 of 270 (33.7%) and 41 of 257 (16.0%) were positive among CAP cases in 2004 and 2005, respectively. More positive cases were seen between spring and early summer and in cases aged 70 years or more, especially those over 80 years old. These results indicated that further evaluation is required among positive cases in elder group. PMID- 17419435 TI - [Prognostic significance of neutrophil elastase inhibitor in patients with acute lung injury and interstitial pneumonia]. AB - The prognostic significance of a neutrophil elastase inhibitor, sivelestat sodium hydrate (SSH), was evaluated in patients on mechanical ventilation due to acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) associated with systemic inflammatory response syndrome (SIRS). We studied 20 consecutive patients in our hospital, where patients complicated with interstitial pneumonia (IP) were separately analyzed (ARDS group, n = 10; IP group, n = 10). There was no significance difference between the two groups in the average lung injury score (3.0 in the ARDS group versus 2.8 in the IP group), the mean P/F ratio (96.3 mmHg in the ARDS group versus 96.7 mmHg in the IP group), plateau pressure (30.5 mmHg in the ARDS group versus 27.0 mmHg in the IP group), ventilator-free days, the duration of intensive care unit stay, and the hospitalization period. Four ARDS patients and 5 IP patients were treated with SSH within 3 days from SIRS onset. In the ARDS group, 5 patients (50%) were complicated with 4 or more organ dysfunctions and 3 patients (30%) died. IP patients all received corticosteroid, but the mortality was significantly higher among patients with IP than among those with ARDS by Kaplan-Meier survival curves. Of the clinical variables, only the diagnosis of IP was found to be independently related to mortality by a multivariate Cox proportional-hazards analysis. We conclude that IP patients have poor life expectancy if they are treated with SSH. PMID- 17419436 TI - [A case of malignant pleural mesothelioma with elevation of G-CSF and CYFRA in the serum and pleural fluid]. AB - A 68-year-old man complaining of hoarseness and back pain, with no history of exposure to asbestos, was referred to our hospital in June 2002. He was admitted because his chest X-ray and CT scan showed atelectasis and a tumor-like region in the right lower lobe of the lung. Serum-CYFRA was 2.8 ng/ml, elevated slightly; however, no other tumor markers for lung cancer were elevated. A diagnosis of squamous cell lung cancer was made based on bronchial washing cytology. Persistent high fever and WBC count elevation did not respond to antibiotics, and reduced only after chemotherapy. Both serum G-CSF (217.0 pg/ml) and CYFRA in the pleural effusion (107.1 ng/ml) were elevated. The biopsy of the growing tumor in the right lateral abdominal wall revealed carcinoma with sarcomatous component or biphasic-type malignant pleural mesothelioma (MPM). In spite of chemotherapy and radiation therapy for the abdominal wall tumor, the tumor rapidly progressed and the patient died three months after admission. The findings at autopsy suggested the tumor was a sarcomatous MPM. However, immunohistochemical staining and tissue HABP staining revealed biphasic type MPM. Although CYFRA elevation in the serum and/or the pleural effusion in MPM patients has been previously reported, it has not been reported in any of the 5 MPM patients reported to have G-CSF elevation. Therefore, this is the first reported case of G-CSF-producing MPM with CYFRA elevation in both serum and the pleural effusion. PMID- 17419437 TI - [A case of nonspecific interstitial pneumonia with reversed halo sign on chest HRCT]. AB - A 39-year-old man was referred to our hospital with anterior chest discomfort, dry cough and shortness of breath. His blood test revealed mild inflammatory change and high serum KL-6 levels. Chest radiograph and computed tomography (CT) showed ground glass attenuation with volume loss in both lower lung fields, and in particular a reversed halo sign was shown on high-resolution CT (HRCT). As transbronchial lung biopsy and bronchoalveolar lavage did not enable a diagnosis, video-assisted thoracic surgery was performed. The histological findings of the resected specimen showed cellular nonspecific interstitial pneumonia. This suggested the possibility of collagen vascular disorder (CVD) associated with interstitial pneumonitis, but no criteria of CVD were fulfilled. Although the reversed halo sign is relatively specific for cryptogenic organizing pneumonia, we report a case of cellular nonspecific interstitial pneumonia showing this sign on chest HRCT. PMID- 17419438 TI - [Two familial cases of psittacosis]. AB - We report here 2 cases of psittacosis in a family. In the first case, a 51-year old woman was admitted with fever, dry cough, and a chest radiograph showed increased opacity in the right upper lung field. On a diagnosis of atypical pneumonia, minocycline was given and her clinical symptoms and abnormal laboratory data were improved. The second case was her husband, a 58-year-old man who presented with fever 4 days after his wife's admission. His chest radiograph revealed increased opacity in the left lower lung field. The administration of azithromycin for 3 days attenuated his clinical symptoms and his abnormal laboratory data improved. The serum titer of complement fixation (CF) test and ELISA test against Chlamydophila psittaci were elevated in both cases on analysis of paired acute- and convalescent-phase serum speciments. The antigen of Chlamydophila was revealed from these parrots, which had been raised in their family. Therefore, we concluded that the psittacosis had originated from the parrots. PMID- 17419439 TI - [A case of drug-induced pleuritis, possibly due to Hochuekkito]. AB - We report a case of drug-induced pleuritis possibly due to Hochuekkito. Hochuekkito, a Chinese-Japanese herbal medicine, was prescribed for a 33-year-old woman with panic disorder and depressive state. About 10 weeks after the first administration, she was admitted complaining of chest pain and cough. Chest X-ray and computed tomography showed the right pleural effusion. Thoracocentesis revealed eosinophilic pleuritis. Discontinuation of Hochuekkito resulted in improvement of the clinical findings. Based on the clinical course, we considered this case to be Hochuekkito-induced pleuritis. PMID- 17419440 TI - [A case of Wegener's granulomatosis presenting as severe acute respiratory failure]. AB - A 19-year-old man was admitted to another hospital. Pulmonary suppuration was diagnosed and was treated with antibiotics. However, he developed acute respiratory failure, which required intubation and ventilation with 100% oxygen. After treatment with nitric oxide inhalation and corticosteroid pulse therapy, the patient's condition stabilized and he gradually regained a satisfactory pulmonary function. He was discharged about 3 months after admission with a pulmonary function close to normal. Approximately 1 month later, the patient was admitted to our hospital because of a 2-week history of fever and chest and ocular pain. A chest radiograph obtained upon admission showed a nodule with a cavity in the upper lobe of the right lung. Pulmonary suppuration was again suspected, and antibiotics were given. The fever persisted and chest radiograph on hospital day 19 showed marked extension of the nodules. At that time, the patient complained of nasal obstruction and hoarseness and his sclera showed intense congestion, indicating episcleritis. Wegener's granulomatosis was diagnosed on the basis of the clinical picture, PR3-ANCA titer (63 EU) and nasal biopsy findings. After treatment with prednisolone and cyclophosphamide, his condition stabilized, and he recovered gradually. However, his condition remains poor despite continued therapy. This is an extremely rare case of Wegener's granulomatosis presenting as severe acute respiratory failure. PMID- 17419441 TI - [A case of meningeal carcinomatosis from lung cancer successfully treated with aggressive multimodal therapy]. AB - A 44-year-old man was admitted to our hospital because of brain metastasis and intrapulmonary exacerbation of lung adenocarcinoma. Systemic chemotherapy (PAC + CBDCA) was administered, but neurological symptoms (muscle weakness of limbs and disorientation) appeared. Lumbar puncture and enhanced MRI of lumber vertebrae revealed meningeal carcinomatosis. MTX 20 mg/week (+ Ara-C 40 mg/week) was injected into the meningeal space. There was a clear improvement in the neurological symptoms, but it did not last long. Meningeal injection was performed 7 times. Whole-brain and whole-marrow radiation along with systemic chemotherapy gave a marked improvement in the symptoms and in the metastatic shadow on the chest CT scan. PMID- 17419442 TI - [A case of juvenile lung cancer with suspected progeroid syndrome and mental retardation]. AB - A 28-year old woman developed juvenile lung cancer. Cockayne syndrome, one of the progeroid syndromes, was suspected because of mental retardation, renal disfunction, photosensitivity, and the characteristic physical appearance such as low set ears, microcephaly, senile face, short stature, and cachectic habitus. We tried chemotherapy with gefitinib and docetaxel, but they were ineffective and she was found to have multiple extraocular muscle metastasis. CT scan showed carcinomatous lymphangiosis findings, her respiratory condition worsened gradually and she died about 7 months after the diagnosis of lung cancer. Some progeroid syndromes are known for their high incidence of juvenile cancres, and the responsible genes are gradually coming to light. However, concerning the relationship between lung cancer and progeroid syndromes, our investigations revealed only one case report of Werner syndrome, one of the progeroid syndromes, accompanied by lung cancer. Progeroid syndromes with lung cancer are thought to be very rare. We expect collection of data on cases like the present case will help to clarify the mechanism of aging and carcinogenesis. PMID- 17419443 TI - [Early stage pulmonary alveolar proteinosis detected by chest CT scan in a medical examination]. AB - We report a case of pulmonary proteinosis detected at an early stage and followed up on chest CT. A 49-year-old man underwent detailed examinations because of abnormal shadows on chest CT taken on a routine medical examination. The chest CT revealed almost symmetrical ground glass opacities (GGOs) in both lungs with thickened alveolar septa. We could not make a definitive diagnosis even with bronchoalveolar lavage and transbronchial lung biopsy, but after about half a year, the GGOs increased. VATS-biopsy demonstrated alveoli filled with PAS positive granular materials, and we made a diagnosis of pulmonary alveolar proteinosis. This case was found at an early stage and we were then able to follow up the disease. PMID- 17419444 TI - [Planned neck dissection after weekly docetaxel and concurrent radiotherapy for advanced oropharyngeal cancer]. AB - Small oropharyngeal carcinomas with advanced neck metastases (stage N2 or greater) are common. Patients with small T with large N oropharyngeal carcinoma have high rates of local control but lower rates of regional control when treated with chemoradiotherapy. Clinical assessment after chemoradiotherapy cannot ensure the absence of neck disease. In the last 5 years, we have treated patients with T1-2 with N2-3 oropharyngeal carcinoma with weekly docetaxel radiotherapy followed by planned neck dissection (PND). Our objectives were to clarify the pathologically complete response (CR) rate of neck metastasis after weekly docetaxel radiotherapy, to identify the clinical predictor of residual neck disease, and to determine the mobidity of planned neck dissection. After chemoradiotherapy, all 12 patients had a complete response at the primary site. We conducted 15 neck dissections. Of these, 6 (40%) had positive nodes. The pathological CR rate of neck metastasis was 58.3%, whereas overall 2-year neck control rate was 91.7%. These findings lend support to the role of PND after chemoradiotherapy in N2-3 neck disease. After chemoradiotherapy, clinical parameters including TN status, feasibility of chemoradiotherapy, largest lymph node size or size reduction in MRI, did not identify patients with residual neck disease. We conducted selective neck dissection (SND) in 80% of patients. SNI) as PND appears to be appropriate in this group of patients because of the low incidence of complications. A further cohort study including the comparison of PND nonenforcement group is necessary to clarify the validity of the addition of PND in weekly docetaxel radiotherapy. PMID- 17419445 TI - [Clinical study of neck dissection for lymph node metastasis in patients with malignant skin tumors of the forehead and face]. AB - We reviewed the records of 10 patients with malignant skin tumors of the forehead and face who underwent neck lymph node dissection at Kurume University Hospital between 2000 and 2004. Two patients underwent selective neck dissection (SND), 5 patients underwent SND and superficial parotidectomy (SP) and 3 patients underwent modified radical neck dissection and SP. Lymph node metastasis to the upper jugular group was found in 3 patients, and metastasis to lymph nodes of the parotid region was found in 3 patients. In a patient with malignant melanoma of the forehead, the patent blue dye was injected intradermally around the tumor and blue-stained lymph nodes were identified in the upper jugular group and parotid region. From these results, we consider that the sentinel lymph nodes of frontal and facial malignant tumors are located in the upper jugular group and parotid region. Thus, in malignant skin tumor patients, SND and SP might be mandatory. PMID- 17419446 TI - [Acute onset Wegener's granulomatosis presenting as otitis media; a case report]. AB - We report the case of a 57-year-old woman with Wegener's granulomatosis who presented with otitis media. The patient presented with a 2-month history of bilateral hearing loss and dizziness. Antibiotic treatment was not effective, and the patient was confirmed to have bilateral sensorineural hearing loss. Serum was positive for cytoplasmic anti-neutrophil cytoplasmic antibody [C (PR3)-ANCA] but negative for P(MPO)-ANCA (perinuclear ANCA). While the test results were pending, the patients' general condition worsened ; in particular, signs of active inflammation, cardiorespiratory failure and scleritis developed suddenly. Emergency chest computed tomography revealed evidence of lung involvement and hydrothorax; thus, the patient was diagnosed to have the generalized form of Wegener's granulomatosis. The patient was started on pulse methylprednisolone therapy at 1000mg/day for 3 days, which resulted in marked clinical improvement, and then, the drug therapy was switched to prednisolone 60mg/day and cyclophosphamide 50mg/day and gradually tapered. Early diagnosis of Wegener's granulomatosis is often difficult because of atypical manifestations of the disease; particular attention must be paid to acute onset of the disease, such as in our case. ANCA is a very useful marker for early diagnosis, but about one week is needed to obtain the test results. We believe that early steroid and cyclophosphamide therapy is an effective therapeutic option for patients with signs of severe inflammation and generalized involvement. PMID- 17419447 TI - [Nanoscience in rotary molecular motor proteins]. PMID- 17419448 TI - [Single molecule study on the mechano-chemical coupling of F1-ATPase motor by the use of an ultra small reaction chamber]. PMID- 17419449 TI - [Single molecule technique on the rotation and function in the supra-molecular motor of flagella]. PMID- 17419450 TI - [Advanced method for single molecule imaging: application to rotary molecular motor]. PMID- 17419451 TI - [Membrane protein analysis with MEMS technologies]. PMID- 17419452 TI - [Analysis of structure and function of F1-ATPase molecular motor protein studied by molecular simulations]. PMID- 17419453 TI - [Unveiled multifunctionality of V-ATPase and the molecular mechanism revealed by X-ray crystal structures]. PMID- 17419454 TI - [Regulation of ribosome biogenesis by the protein kinase TOR in response to nutrient availability]. PMID- 17419455 TI - [Novel chemical mediators in the resolution of inflammation]. PMID- 17419456 TI - [Spermatogenic stem cells and pluripoent stem cells]. PMID- 17419457 TI - Interpersonal performance. PMID- 17419458 TI - Practical management of common atrial arrhythmias 1: cardioversion. AB - This is the first in a series of four practical articles highlighting the important management steps for non-cardiologists and non-cardiac electrophysiologists dealing with patients with atrial fibrillation and common atrial flutter. This article will deal with care pathways and management principles for cardioversion of atrial arrhythmias. PMID- 17419459 TI - Doppler cardiac output monitoring: a tool for all physicians. PMID- 17419460 TI - Ambulatory cardiac rhythm monitoring. AB - Cardiac arrhythmias may cause palpitations, dyspnoea, angina pectoris, dizziness or even syncope and sudden death. This article will review the indications for investigation, the novel devices that are available for investigating patients with suspected cardiac dysrhythmias and the new technology available for analysis and reporting. PMID- 17419461 TI - Principles and procedures of medical ethics case consultation. AB - Ethical conflicts are common in hospital medicine. This article reviews core medical ethics principles, describes models for conducting hospital-based ethics case consultations, and highlights the contributions of hospital ethics committees to high quality patient care. PMID- 17419462 TI - Junior doctors' hours and pay: a guide for foundation year doctors. PMID- 17419463 TI - Acute breathlessness. PMID- 17419464 TI - The applied anatomy of chest drain insertion. PMID- 17419465 TI - Spinal immobilization for the junior doctor. PMID- 17419466 TI - The Mental Capacity Act. PMID- 17419467 TI - Medical causes of abdominal pain. PMID- 17419468 TI - The management of constipation in hospital inpatients. AB - This article reviews the causes of constipation in hospital and how it can be prevented with simple measures. A review of laxatives available on hospital words is provided for the reader and recommendations are made. PMID- 17419469 TI - Bone bruises: definition, classification and significance. AB - The increasing use of magnetic resonance imaging in the diagnosis of musculoskeletal injuries has alerted clinicians to the phenomenon of bone bruising. This article will outline the current knowledge and treatment implications of these interesting radiological findings. PMID- 17419470 TI - Detention on a general ward: understanding and applying the law. AB - From April 2007 there will be two acts of parliament that govern the legal detention of patients in hospitahl: the Mental Health Act 1983 and the Mental Capacity Act 2005. This article addresses common questions posed to liaison psychiatrists by hospital doctors: how to legally detain patients on general wards and how these Acts are applied. PMID- 17419471 TI - How much do new junior doctors in emergency medicine understand about oxygen therapy? AB - Oxygen is routinely prescribed in hospital practice. It should be prescribed like any other drug, specifying delivery device, flow rate and desired inspired concentration. There appears to be continued deficiency in the provision of guidance for, and understanding of, the principles of oxygen therapy. PMID- 17419472 TI - 'Just a sharp scratch': permanent radial, median and ulnar neuropathy following diagnostic venepuncture. PMID- 17419473 TI - Ectopia lentis caused by blunt eye trauma. PMID- 17419475 TI - Physician test thyself. PMID- 17419474 TI - Doctors and demons. PMID- 17419476 TI - A-day 1 year on. PMID- 17419477 TI - Differences in mental health law training. PMID- 17419478 TI - Major elective gastrointestinal surgery: does fluid restriction improve outcome? PMID- 17419479 TI - [Severity levels of intraoperative injury in patients with diffuse disseminated pulmonary syndrome]. AB - Differential diagnosis of diseases manifested with lung diffuse dissemination syndrome is very difficult. Final diagnosis is possible only after pathologic examination of specimens taken during diagnostic surgical procedures. Results of open and videothoracoscopic lung biopsy at 107 patients are analyzed. Assessment of surgical trauma was based on respiratory function, neuroendocrine system response and doses of analgesics in postoperative period. PMID- 17419480 TI - [Prediction of respiratory complications after radical surgeries for non-small cell carcinoma of the lung]. AB - A predictive role of volume-velocity (VV) indices of ventilation in prognosis of respiratory complications after radical surgeries for non-small cell carcinoma of the lung was evaluated. It is demonstrated that decreased VV indices of ventilation before surgery directly correlate with a respiratory complication rate after surgery. New methods of diagnosis of ventilation disorders and of monitoring are regarded as promising. PMID- 17419481 TI - [Invasive treatment of acute coronary syndrome in multiple affection of the coronary arteries]. AB - A comparative assessment of the risk of invasive myocardial revascularizations in patients with multiple affection of coronary arteries and acute coronary syndrome (ACS) without stable ST-elevation on ECG was carried out. Group A (n=45) consisted of patients with one-stage revascularization with coronary bypass (CB) during ACS, group B (n=45) - balloon angioplasty /stenting of symptom-determining artery with subsequent CB in the "cold period" (6 months after ACS episode). It is demonstrated that the one-stage procedure (i.e. multiple CB during ACS) is more effective compared with two-stage invasive treatment because of lower lethality and lower rate of CHD complications. PMID- 17419482 TI - [Efficacy of surgical and therapeutic methods of repeated carotid ischemic stroke prophylaxis]. AB - Comparative efficacy of surgical and therapeutic methods of repeated ischemic stroke prophylaxis at the patients with clinical symptoms of acute brain ischemia and carotid stenosis less 70% was analyzed. Three-year period of follow-up demonstrated that at the patients who have undergone carotid endarterectomy the repeated ischemic events were seen only at 4 (3.9%), and in the group of therapeutic prophylaxis--at 57 (52.7%). Carotid endarterectomy permits to reduce relative risk of repeated ischemic stroke from 0.77 in therapeutic prophylaxis to 0.07 in surgical treatment. Antiaggregant therapy after surgery is mandatory. Prolonged administration of Clopidogrel permits to reduce restenosis and thrombotic complication rate. PMID- 17419483 TI - [Why is the gender of the patient important in prediction of recurrent duodenal ulcer bleeding?]. AB - Results of treatment of 308 patients under 60 years of age with verified stopped ulcer duodenal bleeding were analyzed retrospectively. The number of recurrent bleedings among 85 women was less than among 223 men with similar location of ulcer, severity of blood loss and methods of treatment. Basing on the results of treatment of 99 patients who received no transfusions of blood components it was demonstrated that the velocity of erythrocytes and hemoglobin natural recovery after blood loss was significantly higher among women than among men. PMID- 17419484 TI - [Endoscopic procedures in the treatment of residual and recurrent choledocholithiasis]. AB - Results of endoscopic treatment of 124 patients with residual and recurrent choledocholithiasis after cholecystectomy were analyzed. It is demonstrated that endoscopic papillosphincterotomy with lithoextraction is the method of choice. Lithoextraction after mechanical lithotripsy is indicated in different sizes of the concrement and terminal part of the common bile duct. Conventional nasobiliary drainage (NBD) in incomplete evacuation of the concrements not always predicts repeated impaction of the stone. Original spiral-type catheter procedure for NBD permits one to prevent repeated impaction of the concrement and to perform operation after attenuation of acute purulent cholangitis. PMID- 17419485 TI - [Diagnosis and treatment of acute amebian liver abscess with laparoscopic method]. AB - Laparoscopic surgical treatment was performed at 55 patients with acute amebian liver abscess. The diagnosis of acute amebian liver abscess was verified with clinical, serological, echoscopic, x-ray and CT methods. The laparoscopic method permits to sanify the abscess cavity of any localization. All the 55 patients have been discharged in good health. Laparoscopic method is maximally rational, associated with minimal surgical trauma, permits to decrease hospital stay and period of postoperative rehabilitation. PMID- 17419486 TI - [Erythropoietin use in extensive surgeries on liver and pancreas]. PMID- 17419487 TI - [Treatment of giant postoperative ventral hernia complicated with eventration and small intestine rupture]. PMID- 17419488 TI - [Extraabdominal localization of appendix mucocele]. PMID- 17419489 TI - [Multistage treatment of esophagus and stomach total severe burn]. PMID- 17419490 TI - [Severe complication of acute appendicitis]. PMID- 17419491 TI - [Protamin-sulfate in cardiac surgery: between Scilla and Haribda]. PMID- 17419492 TI - [Cell therapeutic technologies of a new generation: preconditions and problems of realization]. PMID- 17419493 TI - [New developments in the problem of chronic tonsillitis]. PMID- 17419494 TI - [Contribution of the otorhinolaryngology chair of the Dagestan Medical Academy to the study of the diseases of the upper respiratory tract in respect of their relations with nonspecific bronchopulmonary pathology]. PMID- 17419495 TI - [Impact of vibration on impulse activity of neurons of ipsilateral medial vestibular nucleus of unilaterally delabyrinthined rats]. AB - Background impulse activity of the neurons of ipsilateral medial vestibular nucleus of unilaterally delabyrinthined rats was studied on different days after operation as well as after 5-day vibration. It was found that in intact animals mean neuron discharge rate is 16.6+/-1.7 Hz. On day 2 after affection it falls to 14.7+/-0.9 Hz and returns to the baseline on day 7 after surgery (16.6+/-1.2 Hz). Vibration impact raises mean impulse frequency to 21.2+/-2.8 Hz. Characteristics and significance of the results obtained are discussed. PMID- 17419496 TI - [Functional impairment of acoustic and vestibular analyzers in patients with blast and traumatic head injuries]. AB - A comprehensive otoneurological investigation of 131 patients with head injuries of shoch-wave and mechanical nature has revealed similar acoustic, vestibular and central hemodynamic disorders in the above patients. The diagnosed pathological features are determined only by strength and extention of the traumatic factor but not by its nature. Common features in development of acoustic and vestibular disorders in patients with different head injuries enable design of an integrated therapeutic and diagnostic algorithm of management of the above patients with the drugs tanakan and gincor fort. PMID- 17419497 TI - [The role of the otoneurological method in early diagnosis of multiple sclerosis]. AB - Complete vestibulometric, audiological and otoneurological examinations were made in 48 patients with multiple sclerosis (MS). Central cochlear-vestibular syndrome was diagnosed in all the examinees. The otoneurological examination and comparison of changes in the acoustic and vestibular functions revealed specific symptoms characteristic for demyelinating disease of the central parts of the cochleovestibular analyzer. MR-tomography visualization of the demyelination foci in the brain confirmed affection of the cochleovestibular analyzer. PMID- 17419498 TI - [Acoustic function in patients early after minor craniocerebral trauma]. AB - Acoustic function was studied in 101 patients early after closed craniocerebral trauma (CST). In acute CST hearing impairment depends on trauma severity: severe hearing loss occurs in severe CST. In mild CST hearing defect depends on trauma location: serious hearing loss is observed in occipital trauma. Diagnostic examination in acute CST should include audiometric investigation. Documentation of hearing loss degree and its dynamics is important for labor and forensic medical expert evaluation. PMID- 17419499 TI - [Clinical symptoms of branchio-oto-renal syndrome in a family with a positive test for EYA1 gene]. AB - Three families with branchio-oto-renal syndrome (BOR syndrome) were examined. In one of the families all its members with BOR syndrome had deletion of cytosine in position 759 (759delC) in DNA sequence of EYA1 gene in exone 8. Clinical characteristics of BOR syndrome in the family are given. Molecular-genetic analysis confirmed the diagnosis clinically in case of all signs of the syndrome presence among the members of the family. Rare cases of hereditary syndromes with hearing problems provide more knowledge about structure of hereditary hypoacusis forms in the population. The syndromal forms reflect a complex genetic basis of the processes of sound perception. PMID- 17419500 TI - [Treatment of children with chronic adenoiditis in different coagulation status]. AB - Chronic adenoiditis in children is a frequent surgical pathology. To avoid hemorrhagic complications in adenotomy, it is necessary to exclude concomitant hemostatic pathology. Forty-three children were examined before operation. Primarily, vascular-thrombocytic disorders (dysaggregation thrombocytopathies) were found. Chronic focal infection plays an essential role in pathogenesis of systemic disorders in chronic adenoiditis including those provoking coagulatory disorders. A program of a target pre- and intraoperative hemostatic therapy is proposed which has demonstrated its high efficacy. PMID- 17419501 TI - [Treatment of allergic rhinitis associated with hypothyroidism]. AB - Fifty five patients with allergic rhinitis and moderate hypothyroidism due to autoimmune thyroiditis have undergone clinical, functional, hormonal, immunological and allergological examinations. Of them, 45 patients received a course of treatment with thyroid hormones and antiallergic drugs while 10 patients received replacement hormonal therapy only. The results of the study demonstrate that treatment of allergic rhinitis associated with hypothyroidism is more effective if antiallergic drugs are added to replacement hormonal therapy. PMID- 17419502 TI - [Comparative efficacy of steroid drugs in the treatment of all-the-year-round allergic rhinitis]. PMID- 17419503 TI - [Implication of middle-mass molecule peptides of blood plasma in assessment of intoxication syndrome in patients with bacterial tonsillitis]. AB - The level of middle-mass molecule peptides (MMMP) of blood plasma was investigated in 98 patients with tonsillitis. A significant increase in the level of MMMP with a maximal rise at the height of the disease with normalization in early and late convalescence was revealed. The changes in plasma MMMP in tonsillitis depended on the clinical form, stage and severity of the disease, complications, concomitant diseases, completeness of convalescence. High therapeutic efficacy of an immunomodulating drug tamerit in combined therapy of patients with bacterial tonsillitis is shown. PMID- 17419504 TI - [Individual characteristics of the shape and structure of the nasopharynx related to structural characteristics of the base of the skull and facial skeleton in children]. AB - Examination of 60 skulls of the children aged 1 to 16 years has established that the size of the choanae determines width and height of the nasopharynx while width of the choanae correlates (r=0.8) with the distance between the inferior turbinated bones. There is also a significant correlation between the shape of the facial vault and the vault flexure. PMID- 17419505 TI - [Correction of nasal breathing in combined treatment of respiratory pathology]. PMID- 17419506 TI - [Long-term surgical outcomes in congenital malformations of the floor of the auricle]. AB - The authors analyse long-term outcomes of surgical treatment of congenital defects of the floor of the auricle. They came to the conclusion that surgical techniques used in the correction of the defects need some modification. In the presence of a wide, elastic ascending crus of the helix, when creating the anthelix, the crus should be made finer. In the presence of a large and wide scaphoid fossa it should be made smaller. After reduction of the floor of the auricle the margin of the auricle should be shortened. PMID- 17419507 TI - [Safety of antibiotic therapy of sinusitis]. PMID- 17419508 TI - [Therapy of imudon after tonsillectomy]. PMID- 17419509 TI - [Nonpharmalogical correction of respiratory disorders in poor health children]. PMID- 17419510 TI - [Efficacy of perioperative prophylaxis with bioparox in surgical interventions on the nasal cavity and paranasal sinuses]. PMID- 17419511 TI - [Optimal decision on analgetic therapy in patients after tonsillectomy]. PMID- 17419512 TI - [Anti-inflammatory therapy of paranasal sinuses diseases]. PMID- 17419513 TI - [Variants of the structure of the stylohyoid complex]. PMID- 17419514 TI - [Cure of epilepsy after sanation of the paranasal sinuses]. PMID- 17419515 TI - [Location of the impacted tooth in the maxillary sinus]. PMID- 17419516 TI - [Unusual foreign bodies of the pharynx and esophagus]. PMID- 17419517 TI - [Updated treatment methods and policy in otitis media purulenta and its complications]. PMID- 17419518 TI - Adductor spasmodic dysphonia versus muscle tension dysphonia: examining the diagnostic value of recurrent laryngeal nerve lidocaine block. AB - OBJECTIVES: Differentiating adductor spasmodic dysphonia (ADSD) from muscle tension dysphonia (MTD) can be difficult. This investigation examined the precision of response to unilateral lidocaine block of the recurrent laryngeal nerve (RLN block) as a potential diagnostic test to discriminate ADSD from MTD. METHODS: Patients with ADSD (n = 23) and MTD (n = 20) were audio-recorded before and during RLN block. The patients completed self-ratings of dysphonia severity, vocal effort, and laryngeal tightness, and blinded listeners completed auditory perceptual ratings of overall severity, breathiness, and strain of voice samples before and during the block. RESULTS: Repeated-measures analysis of variance, with "group" (ADSD/MTD) as the between-subjects variable and "time" (before block/during block) as the within-subjects variable, confirmed significant "time" effects, but no significant "group-by-time" interaction effects, indicating that both disorder groups responded favorably to RLN block, according to patient- and listener-based ratings. Furthermore, low estimates of sensitivity and specificity and weak receiver operating characteristic curves confirmed that a positive response to the RLN block test did not distinguish ADSD from MTD. CONCLUSIONS: We conclude that RLN block offers little discriminatory value in the differential diagnosis of ADSD versus MTD, and a positive response to RLN block should not be considered confirmatory of ADSD. PMID- 17419519 TI - Immunocompetency in children with recurrent respiratory papillomatosis: prospective study. AB - OBJECTIVES: We sought to investigate the immunologic status of children with recurrent respiratory papillomatosis and to evaluate possible correlations between the patients' immunocompetency and the clinical course of the disease. METHODS: Twenty children with recurrent respiratory papillomatosis underwent immunologic evaluation every 6 months for determination of complete blood count, serum immunoglobulin levels, lymphocyte subpopulations, lymphocyte response to mitogen stimulation, and natural killer cell function. The patients were observed prospectively (42 to 56 months), and their clinical course was recorded. The findings were compared with those in healthy age-matched controls. RESULTS: The CD4/CD8 ratio and the lymphocyte response to mitogen stimulation were significantly reduced in the study children compared to normal controls. A reduction in lymphocyte response to mitogen stimulation was significantly correlated to a high number of papilloma sites and more frequent recurrences. Abnormal natural killer cell function was significantly correlated to more frequent recurrences. CONCLUSIONS: A compromised cell-mediated immune response may be associated with repeated or persistent human papillomavirus infections, leading to the development of recurrent respiratory papillomatosis. Patients with an aggressive clinical course may have underlying cell-mediated immunodeficiency. Long-term prospective investigations are needed to establish the role of the host immune system in the pathogenesis of recurrent respiratory papillomatosis in children. PMID- 17419520 TI - Videokymography in voice disorders: what to look for? AB - OBJECTIVES: Kymographic imaging through videokymography has been recognized as a convenient, novel way to display laryngeal behavior, yet little systematic research has been done to map the relevant features displayed in such images. Here we have aimed at specification of these features to enable systematic visual characterization and categorization of vocal fold vibratory patterns in voice disorders. METHODS: A cross-sectional, descriptive design was used. We selected 45 subjects and extracted 100 videokymographic images from the archive of more than 7,000 videokymographic examinations of subjects with a wide range of voice disorders. The images showed a large variety of vocal fold vibratory behaviors during sustained phonations. We visually identified the prominent features that distinguished the vibration patterns across the images. RESULTS: We divided the findings into 10 feature categories. They included refined traditional features (eg, mucosal waves), as well as additional features that are obscured in strobolaryngoscopy (eg, different types of irregularities, left-right frequency differences, shapes of lateral and medial peaks, cycle aberrations). CONCLUSIONS: The variations in the identified features reveal different behavioral origins of voice disorders. The findings open new possibilities for objective documentation and for monitoring vocal fold behavior in clinical practice through kymographic imaging. PMID- 17419521 TI - Histopathology of ossicular grafts and implants in chronic otitis media. AB - OBJECTIVES: We describe the histopathology of ossicular grafts and implants so as to provide insight into factors that may influence functional results after surgery for chronic otitis media. METHODS: Histopathologic observations were made on 56 cases: 50 surgical specimens and 6 temporal bone cases in which the graft was sectioned in situ. RESULTS AND CONCLUSIONS: Autogenous malleus, incus, and cortical bone grafts behaved in a similar manner and maintained their morphological size, shape, and contour for extended periods of time, at least up to 30 years. These histopathologic observations support the continued use of autograft ossicular and cortical bone grafts for middle ear reconstruction. Cartilage grafts developed chondromalacia with resulting loss of stiffness and showed a tendency to undergo resorption. Synthetic prostheses made of porous plastic (Plastipore, Polycel) elicited foreign body giant cell reactions with various degrees of biodegradation of the implants. Prostheses made of hydroxyapatite and Bioglass were enveloped by a lining of connective tissue and mucosal epithelium. The Bioglass material was broken down into small fragments and partially resorbed by a host response within the middle ear. These results warrant caution in the use of prostheses made of porous plastic or Bioglass. PMID- 17419522 TI - Endoscopic management of blunt pediatric laryngeal trauma. AB - OBJECTIVES: Blunt laryngeal trauma in the pediatric population is an uncommon but unique entity that can be potentially life-threatening. Given the infrequency of these events, its management can pose a clinical dilemma. The authors review the evaluation and treatment of blunt pediatric laryngeal trauma. METHODS: We present a case report and a review of the literature. RESULTS: We describe the case of a 3-year-old boy who presented with laryngeal injury following blunt trauma. The patient sustained endolaryngeal hematomas and mucosal lacerations with exposed cartilage. After mucosal approximation, the patient was successfully managed in a critical care setting without a tracheostomy. CONCLUSIONS: With an appropriate and thorough evaluation of the pediatric patient, endoscopic management without a surgical airway may be considered as a viable alternative for blunt laryngeal trauma. PMID- 17419523 TI - Value of temporal bone density measurements in otosclerosis patients with normal appearing computed tomographic scan. AB - OBJECTIVES: Previous studies demonstrated that otosclerosis diagnosis benefits from temporal bone density measurements. We sought to assess bone density measurements of the fissula ante fenestram (FAF) in normal patients, in patients with otosclerosis, and in patients with cholesteatoma. We discuss the value of temporal bone density measurements in patients with otosclerosis who have a normal-appearing computed tomographic (CT) scan. METHODS: This was a prospective case-control study in which 219 temporal bones (123 adults, 18 to 84 years of age) were included between November 1, 2002, and April 30, 2004. All patients underwent a CT scan of the temporal bones. Axial views were obtained with density measurement of the FAF. RESULTS: The FAF density was significantly different (p < .0001) in the otosclerosis group (n = 119) compared to the control group (n = 100). There was no significant difference between the otosclerosis group with a normal-appearing CT scan and the control group (p = .64). CONCLUSIONS: From our results, it may be suggested that 1) temporal bone density measurements seem not to be strictly comparable between CT scan devices; and 2) temporal bone density measurements of the FAF did not allow the diagnosis of otosclerosis when the CT scan appeared normal. PMID- 17419524 TI - Height and shape of the skull base as risk factors for skull base penetration during endoscopic sinus surgery. AB - OBJECTIVES: Computed tomography (CT) and magnetic resonance imaging have identified several risk factors for life-threatening complications of skull base penetration during endoscopic sinus surgery (ESS). We compared these risk factors between groups of patients with and without penetration. METHODS: We performed a retrospective review of direct coronal paranasal sinus CT scans. Using preoperative CT scans of 100 patients without and 7 patients with penetration, we classified height into 4 groups and contour into 2 groups. The frequencies of shape and height differences of the right and left halves of the skull base were calculated in each group. RESULTS: In 6 of the 7 patients who had skull base penetration, ESS was performed by a resident or junior staff member who had less than 3 years of experience with this technique. Shape asymmetry was seen in 4 of the 7 patients (57%) with penetration, which was a significantly higher rate than in patients without penetration (18 of 100; p = .032). The frequencies of a low skull base and a height difference were 15% and 28%, respectively. CONCLUSIONS: The most important risk factor for skull base penetration was the surgeon's inexperience. An asymmetry of shape of the right and left halves of the skull base was significantly related to inadvertent skull base penetration during ESS. PMID- 17419525 TI - Risk factors for adult laryngotracheal stenosis: a review of 74 cases. AB - OBJECTIVES: We sought to identify risk factors for and review our experience in the management of adult acquired laryngotracheal stenosis (LTS) at an academic urban medical center. METHODS: A retrospective review of all patients given a diagnosis of acquired LTS between 1997 and 2005 was performed. Seventy-four patients with LTS were identified. Demographic information, medical and surgical history, surgical procedures performed, and outcomes were collected. A control group of 106 patients admitted over the same time period with respiratory distress but without LTS was identified. The data collected for both groups were analyzed by Fisher's exact test and logistic regression analysis. RESULTS: Demographically, the control group was not significantly different from the LTS group. Patients who had a previous tracheostomy were 10.99 times more likely to develop LTS than control patients (95% confidence interval [CI], 4.68 to 25.80). Patients irradiated for carcinomas of the oropharynx and larynx were 5.95 times more likely to develop LTS than control patients (95% CI, 1.87 to 18.91). Those previously intubated for more than 48 hours were 3.91 times more likely to develop LTS than control patients (95% CI, 1.91 to 8.02). Finally, patients who were intubated for any non-airway surgery were found to be 2.07 times more likely to develop LTS (95% CI, 1.09 to 3.93). CONCLUSIONS: Prolonged intubation, tracheostomy, previous non-airway surgery, and irradiation for oropharyngeal and laryngeal tumors are risk factors for LTS. Multiple surgical procedures are often required for treatment. Mitomycin C did not significantly improve decannulation rates. PMID- 17419526 TI - Endoscopic lysis of anterior glottic webs and silicone keel placement. AB - OBJECTIVES: Acquired anterior glottic webs occur most commonly after endoscopic resection of laryngeal papilloma involving the anterior vocal folds. Treatment of anterior glottic webs has included a tracheotomy with laryngofissure and placement of a laryngeal stent or keel. We have used an endoscopic technique of web lysis and placement of a laryngeal keel without tracheotomy over the past 7 years. METHODS: A retrospective chart review was conducted of all cases of endoscopic web lysis and keel placement performed by the senior author (S.A.B.). RESULTS: Over the past 7 years, 10 patients underwent the procedure, with a mean follow-up of 18 months. The length of the anterior web was up to two thirds of the membranous vocal fold. Outcomes analysis revealed a recurrence in 1 patient and 2 minor complications necessitating treatment. CONCLUSIONS: Endoscopic web lysis and keel placement offers superior results with less morbidity compared to open techniques. PMID- 17419527 TI - Vortical flow field during phonation in an excised canine larynx model. AB - OBJECTIVES: To more fully understand the mechanisms of vocal fold vibration and sound production, we studied the velocity flow fields above the folds. Such velocity fields during phonation have not been reported in the literature. METHODS: Using the particle image velocimetry method for 3 excised canine larynges, we obtained the velocity fields in the mid-membranous coronal plane during different phases of phonation. The velocity field was determined synchronously with the vocal fold motion recorded by high-speed videography. RESULTS: The results show that vortices occur immediately above the vocal folds and that the location and shape of the vortices depend on the phase of the phonation cycle. Consistent vortical structures found included starting vortices, Kelvin-Helmholtz vortices, entrainment vortices, and vortices directly above the folds during the divergent glottal stage. CONCLUSIONS: These vortical structures were consistently found during specific phases of the glottal cycle for 3 canine larynges that significantly varied in size. This consistent behavior suggests that the vortices may be important for both vibration and sound production; however, further study is needed to prove this. The clinical significance of these vortices is discussed. PMID- 17419528 TI - Human lung mucous glands manifest evidence of the H+/K+-ATPase proton pump. AB - OBJECTIVES: The H+/K+-ATPase proton pump has been demonstrated in human laryngeal submucosal glands, and is not solely present in the parietal cells of the stomach. Although proton secretion is present in the lung, a variety of mechanisms have been elucidated. The hypothesis of this study is that the H+/K+ ATPase proton pump is one additional pathway of proton secretion in the human lung. METHODS: Fourteen surgical lung specimens from 10 subjects were retrospectively obtained after approval from our Human Subjects Committee. Banked human stomach tissue was used for comparative positive and negative controls. Sections were immunostained with 2 monoclonal antibodies selectively reactive with alpha or beta subunits of the H+/K+-ATPase proton pump. RESULTS: In the human lung, consistent staining for both subunits was present in the mucous gland cells and ducts in all specimens in which mucous glands were present (6 specimens from 5 subjects). Overall, weak to strong staining was present in focal areas within the multicellular mucous glands. There was only scant focal staining in the respiratory epithelium in 4 specimens. Stomach parietal cells exhibited strongly positive staining for both subunits of the proton pump. There was no staining in stomach cells that were not morphologically consistent with parietal cells. CONCLUSIONS: The H+/K+-ATPase proton pump is present in mucous cells and ducts in the human lung, with some variable expression noted. Proton pump inhibitor pharmacotherapy may have a site of action in the human lung, explaining some of the controversies otherwise attributable to interrelatedness of aerodigestive tract disease. PMID- 17419529 TI - Differential expression of human beta defensin 2 and human beta defensin 3 in human middle ear cholesteatoma. AB - OBJECTIVES: The purpose of this study was to investigate the differential expressions of human beta defensin (hBD) 2 and hBD-3 in human middle ear cholesteatoma epithelium. METHODS: The expressions of hBD-2 and hBD-3 were analyzed by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemical staining. Samples were obtained from 10 patients who underwent middle ear surgery for middle ear cholesteatoma. RESULTS: Real-time RT-PCR and Western blot analysis showed that the messenger RNAs and proteins of hBD-2 and hBD-3 were higher in the cholesteatoma epithelium than in normal external auditory canal skin. In cholesteatoma epithelium, hBD-2 and hBD-3 activities were present in the upper granular layer and in the prickle cell layer, but in the normal skin they were poorly expressed in all layers. CONCLUSIONS: Increased expressions of hBD-2 and hBD-3 in cholesteatoma epithelium suggest that cholesteatoma, a chronic inflammatory state of middle ear keratinocytes, may induce an innate immune response. That the induction of hBD-2 was found to be more intense than that of hBD-3 in cholesteatoma epithelium implies that hBD-2 is the major effector in terms of chronic epithelial inflammatory responses. PMID- 17419530 TI - Mental Health Bill makes uncomfortable progress. PMID- 17419531 TI - 'Having a mental health problem is suddenly sexy'. PMID- 17419532 TI - 'We have got to change the attitude of nurses towards food and nutrition'. PMID- 17419533 TI - The drugs decision. PMID- 17419534 TI - In the line of fire. PMID- 17419535 TI - Progress on the diabetes NSF. PMID- 17419536 TI - Measuring capillary refill time. PMID- 17419537 TI - Caring for patients after mechanical ventilation. Part 2: Nursing care to prevent complications. AB - This article, the second in a two-part series, outlines the importance of providing high-quality nursing care and careful monitoring when patients are discontinued from artificial mechanical ventilation. The range of physical and psychological effects they may experience is discussed, and the role of the nurse in patient monitoring and care during this period is outlined. PMID- 17419538 TI - [The need for experiments using primates from a scientific point of view]. AB - Concerning the public discussion on animal experiments using primates, various research fields are demonstrated where non-human primates are necessary for certain scientific reasons at this time. Non-human Primates are used in Germany mainly in regulatory toxicology and pharmaceutical safety studies.A small amount is disposed in different fields of biological or biomedical basic research. This includes in particular neurosciences and infection research. 2006 New and Old World monkeys were needed in Germany in 2005. No chimpanzees are used anymore as laboratory animals in Germany since many years. Several examples are presented to demonstrate that certain research fields need non-human primates as laboratory animals in the foreseeable future. PMID- 17419539 TI - [Ethical and legal aspects of animal experiments on non-human primates]. AB - Animal experiments on non-human primates give cause for ethical concerns for three reasons (1) the inclusion of "ethical animal protection" in the German Constitution (Article 20a of the "Grundgesetz" GG, 2002) has led to real consequences for the application process with respect to the use of primates for fundamental research; (2) the legal requirements in Europe to ensure animal welfare are currently being tightened and (3) the global problem of the protection of species, especially with respect to the capturing and subsequent sale of primates is still unsolved. As a result of the way humans interpret the term justice (the principle of equality) it was to be expected that great apes, being the animals that most closely resemble humans, would play a key role in the establishment of animal protection laws. In 1997,Great Britain and Ireland made it illegal to conduct experiments on great apes. In 1999, New Zealand went even further and created a kind of basic rights for great apes. In 2003,The Netherlands forbade animal experiments using great apes as did Sweden, which also included gibbons in this ban (which is in line with current taxonomy, which considers gibbons to belong to the family Hominidae). In 2006 Austria forbade experiments carried out on chimpanzees, bonobos, gorillas, orang-utans, and gibbons. Only recently, a state commission on ethics in Switzerland demanded that the Swiss government do the same. And the summer of 2006 saw a debate in Spain on the inclusion of the protection of great apes in the primary goals of the state. Due to the principle of equality, a further extension (both geographically and systemically) of the exclusion of great apes from animal experiments is to be expected. Since Article 20a GG on "ethical animal protection" came into effect on August 1,2002, the regulatory authorities in Germany have the right to independently check and control animal experiments as to their ethical tenability (Administrative Court Giessen, confirmed by the Administrative Court of the State of Hessia) i.e. an authorization for experiments in the area of fundamental research may only be given by the authorities if the "ethical tenability" (according to Section 7 paragraph 3 of the German Law on Animal Welfare) is given. The "ethical tenability" of fundamental research experiments follows other rules than those that pertain to applied research. Fundamental research is granted a low to mid-level tolerance of pain and suffering as being ethically tenable; not tenable are stronger or very high doses of pain and suffering. Actually in summer 2006, a proposal for fundamental research using mon- PMID- 17419540 TI - [Importance of the rearing period for laying hens in alternative systems]. AB - Feather pecking and cannibalism are still major problems in alternative systems for laying hens. Literature and practical experience indicate that unfavourable rearing conditions might be important risk factors for the occurrence of these behavioural disturbances during the laying period. Typical rearing conditions of laying hens from 50 rearing units in Germany and Austria are presented. Obvious risk factors during rearing for feather pecking and cannibalism during the laying period were found. Most flocks were kept under high stocking density (mean: 15 pullets per m' useable area) and some flocks had access to litter only after the second week of life or access to raised perches after the fourth week of life. Plumage condition of pullets and laying hens varied widely in non-beak-trimmed as well as in beak-trimmed flocks. The percentage of pullets with damaged plumage was higher in beak-trimmed than in non-beak-trimmed flocks (medians: 53 % versus 30 %, p = 0,022). In laying hens there was a higher percentage of hens with plumage damage in non-beak-trimmed flocks compared to beak-trimmed flocks (medians: 23 % versus 50 %, p = 0,007). Data analysis will be continued, especially with regard to particular risk factors. PMID- 17419541 TI - [Expressive behaviour to assess the emotional states in horses]. AB - The emotional states such as feelings and emotions are not easy to assess by objective methods in animals as well as in humans. Beside measuring physiological variables an aid to assess the emotional states is the analysis of expressive behaviour of an individual respecting the relating context. Especially developed in mammals, which live in obligatory social i.e. in stable permanent social communities, are facial expression and gesture. Also horses display a differentiated expressive behaviour, which can be observed and analysed by humans. Moreover it could be demonstrated, that in situations of stress the display shown by horses (gesture, facial expression, posture) correlate with corresponding physiological reactions. Thus the expressive behaviour is suitable to assess the emotional states of horses depending on the situation. PMID- 17419542 TI - [Status of law-making on animal welfare]. AB - Since the last report there have been major revisions of laws and ordinances. Deliberations on rules of Community law were also continued. On national level, the Act on the Shoeing of Horses amending the Animal Welfare Act and amendments of animal welfare provisions as well as the Deregulation Act were prepared, some of which have meanwhile entered into force. At legislative level, the work on the ratification laws for the Council of Europe conventions (Strasbourg) was concluded in order to enable Germany to adopt the revisions. They include (1) the European Convention for the protection of animals used for experimental purposes and (2) the European Convention for the protection of animals during international transport. At the level of ordinances, the amendment and extension of the Animal Welfare -Farm Animal Husbandry Ordinance are of vital importance for the sections on pig farming and laying hen husbandry. Another section refers to the husbandry of fur animals, on which an ordinance has been submitted to the Bundesrat (German upper house of Parliament). Deliberations on this issue have been adjourned. Drafts of a circus register were prepared to amend the Animal Welfare Act and to adopt a separate ordinance, and they are being discussed with the federal states and associations. Previously,the rules of Community law in the area of animal welfare were adopted as EC directives which the member states had to transfer in national law. This was done by incorporating them into national laws or ordinances, with non-compliance having to be sanctioned. It is the member states' responsibility to establish sanctions. Yet the Commission has introduced a directly operative animal welfare legislation by adopting EC Regulation 1/2005 on the protection of animals during transport. This means that a national implementation is not required. Nevertheless, the establishment of sanctions continues to be the responsibility of the member states. A special authorisation by the legislator is required to be able to impose sanctions based on directly applicable EC law. This is done via the already mentioned Act on the Shoeing of Horses and amendment. To establish sanctions for this Community legislation, a "Sanctions Ordinance" is currently being discussed by the different departments. This way, a link between directly applicable Community legislation and national sanctions is established. At EC level there are currently discussed (1) the "Animal Welfare Action Plan", (2) a draft directive laying down minimum rules for the protection of chickens kept for meat production and (3) preparations for a revision of the directive on the protection of animals used for experimental purposes have become known due to the preparation of a related impact assessment. At the level of international law, the Council of Europe has concluded its work on Annex A of the convention for the protection of animals used for experimental purposes. With regard to the European Convention for the protection of animals kept for farming purposes, the deliberations on fish and fattening rabbits are being continued. There is a discussion on the technical details of the Transport Convention. Since the first animal welfare conference of the International Office of epizootics (OlE) in February 2004 in Paris, two very comprehensive codes on slaughter of animals and on animal transport were adopted. The inclusion of further animal welfare issues into the OIE work programme will be discussed in the next future. PMID- 17419543 TI - [Practical experiences with the new guidelines "Animal Welfare Aspects for the Organization of Pet Markets"of the German Federal Ministry of Nutrition, Agriculture and Consumer Affairs]. AB - Though being controversially discussed in the fore field the new guidelines "Animal Welfare Aspects for the Organization of Pet Markets" of the German Federal Ministry of Nutrition, Agriculture and Consumer Affairs create a standard supported by the main associations of both animal welfare activists and keepers of animals concerned by this subject. This standard was created as a means of achieving high animal welfare standards at these markets all over Germany. First practical experience with the guidelines show that their strict application can greatly improve conditions for the animals. It remains a challenge for the veterinary authorities however to develop inspection concepts for each market jointly with the organizer. Inspections at each event followed by appropriate executive measures further help to ensure conditions in accordance with German Animal Welfare legislation. Besides the opportunity of making conditions with the obligatory licence of pet merchants should be taken. PMID- 17419544 TI - [Pigeon sport and animal rights]. AB - To begin, a short overview of the organization and the realization of the racing pigeon sport. Some physiological facts, relevant to racing pigeons, will be touched on. Lastly, a focus on the flights, their completion and the problems involved with the, in some cases, high number of lost pigeons. The German Club of Pigeon Breeders, has made improvements but, it is certainly not enough. The topic of "City Pigeons" will be briefed. The final part deals with pertinent animal rights issues, causes of mishaps, and some rectifying possibilities, which are available to the government veterinarian. Special emphasis will be placed on the international uniformity of this issue. The lecture should prove that there is a need for every government veterinarian to become actively involved, because the described problematic has a major effect on a very large number of animals. PMID- 17419545 TI - [Transport of racing pigeons and animal welfare]. AB - A short overview about the transport of the racing pigeon from home to the place of liberation will be given. Some facts related to the transport will be explained. Facts like Number of pigeons in one crate, supplying the pigeons with water and food during transportation and at the place of liberation, the supply of the pigeons with fresh air will be explained. PMID- 17419546 TI - Secretory phenomena in the non-lactating human mammary gland. AB - The normal non-lactating premenopausal human mammary gland has been shown by immunohistochemistry and transmission electron microscopy to secrete a number of antimicrobial peptides such as beta-defensins, the cathelicidin LL37, lactoferrin and adrenomedullin. In addition, the non-lactating gland elaborates a prominent glycocalyx at the apical membrane of the glandular epithelial cells, parts of which are shed into the lumen of endpieces and ducts. This glycocalyx includes the mucins MUC 1 and MUC 4, a strongly Alcian Blue positive palyanionic component and sulfated material stained with Aldehyde Fuchsin. MUC 1 and the Alcian Blue positive material are considered to play an antimicrobial role, too. Lactalbumin and lipid droplets also occur in the non-lactating gland. At the EM-level secretory phenomena operating by exocytosis and by means of the apocrine mechanism have been observed. Cytoskeletal components presumably play a role in apocrine secretion. Apart from secretion at the cellular apex, secretion at the cellular basis also occurs regularly, which may represent the production of para- or endocrine factors. PMID- 17419547 TI - Observations on the actual structural conditions in the stratum superficiale dermidis of porcine ear skin, with special reference to its use as model for human skin. AB - The results obtained from the outer ear skin of female pigs (German Landrace) by light microscopy (LM), transmission electron microscopy (TEM), and cryo scanning electron microscopy (cryo SEM) methods, in particular relying on careful and artefact-free tissue processing, exhibited that the stratum superficiale dermidis of the auricle had a very homogeneous and compact construction, especially in one area (central dorsum auriculae). Based upon the important measurements made [average thickness of stratum superficiale dermidis: 94 (+/-16) microm, region A: 81 (+/-10); average thickness of collagen fibre bundles: 12 (+/-2) microm, region A: 13 (+/-0.5); average density of subepidermal capillaries: 3134 (+/-459) loops/cm2, region A: 3497 (+/-247)], this impression was confirmed by low standard deviations for all parameters, in comparison to marginal locations studied. The capillary system present was analysed by LM and TEM for specific structural features, whereby it generally compared to the microvasculature in human skin. Moreover, a regular pattern of diffusion-relevant punctiform contacts of the capillary loop apex with the epidermal basement membrane became obvious. Cryo SEM, particularly offering the advantages of dispensation of chemical fixation, dehydration and solvents during processing, highlighted delicate structures without shrinkage and without loss of soluble sample components. Thus rather real spatial conditions in the region of the epidermo-dermal junction and the upper dermis were visualized, whereby very regular arrangements of the structures present became obvious. This pertained also to a correct demonstration of all components of the epidermal basement membrane, in particular the lamina lucida. In addition, the water-based stable character of the entire stratum superficiale dermidis could be emphasized as a basic feature for controlled diffusion processes. PMID- 17419548 TI - A co-localization study on the ovine pancreas innervation. AB - The expression of DbetaH and several neuropeptides was investigated in neuronal elements of the ovine pancreas using double immunocytochemical stainings. Immunoreactivities to DbetaH, NPY, VIP and SP were seen to various extents in nerve terminals associated with the acini, islets, ducts, blood vessels, interlobular connective tissue as well as in the neurons of intrapancreatic ganglia. The expression of CGRP was limited to nerve fibers lying in the connective tissue septa, amongst the acini and in close vicinity to the pancreatic blood vessels. Single GRP-positive nerve endings were located around the acini, ducts and in the interlobular connective tissue. With the exception of the ductal system in a co-localization of NPY with DbetaH was frequently found in all regions of the pancreas. Moderately numerous blood vessel-associated VIP positive nerve fibers as well as the vast majority of VIP-containing intrapancreatic neurons were found to co-express DbetaH. Single SP-immunoreactive (IR) nerve fibers of the exocrine pancreas and interlobular connective tissue as well as SP-positive intrapancreatic neurons additionally showed the presence of DbetaH. The co-localization of VIP and NPY was found in nerve terminals located around the blood vessels and acini, in the connective tissue septa as well as in numerous pancreatic neuronal perikarya. Rare nerve terminals located between the acini and around small blood vessels as well as several neurons of intrapancreatic ganglia were VIP-IR/ SP-IR. Simultaneous expression of SP and CGRP was found in nerve fibers supplying large pancreatic arteries and veins, interlobular connective tissue and, occasionally, around the acini. Throughout the pancreas the population of CGRP-positive nerve endings showed lack of VIP and NPY. In a moderate number of GRP-containing nerve fibers, a co-expression of NPY was noted. Nerve terminals containing both GRP and VIP were detected sporadically, whereas none of the GRP-positive nerve terminals showed expression of SP. We conclude that the presented noradrenergic as well as peptidergic innervation patterns of the ovine pancreas are species-dependent. On the basis of the occurrence of DbetaH, NPY, VIP and SP (alone or in combination) in pancreatic neuronal elements we can suggest that these substances presumably act as regulators of the endocrine and/or exocrine pancreas. Involvement of CGRP and GRP in the ovine pancreas physiology seems to be of minor importance. The co localization study indicated that the pancreas of the sheep is innervated from several sources including intrinsic as well as extrinsic ganglia. PMID- 17419549 TI - Confocal laser scanning microscopy of human cementocytes: analysis of three dimensional image reconstruction. AB - In order to define a practical method for a quantitative-qualitative analysis of dental hard tissue cells, we have studied cementocytes of human teeth using Confocal Laser Scanning Microscopy (CLSM), a research technique based on laser light microscopic analysis of biological samples stained for fluorescence observation. One thousand and eight hundred cementocyte images were analyzed. CLSM allowed improved tissue imaging, bi-dimensional pictures with better resolution at cellular level and, in particular, the possibility of a three dimensional image reconstruction, thus providing a dynamic view of the cell under different situations. CLSM allowed a careful morphological observation and dimensional analysis of cementocytes at cellular resolution: cementocyte dimensional parameters like cell body without cell processes, cell dimensions with cell processes, and the number of cellular processes could be obtained. In conclusion, this study reports the fine definition of cementocytes at microscopic level by CSLM and the results warrant the use of this technique for further comparative analyses between normal and pathological cementocyte cells, e.g. cementum neoplasia and periodontal disease which still await a clear analytical description at cellular level. PMID- 17419550 TI - Effect of fluoride ions on apatite crystal formation in rat hard tissues. AB - Fluoride is widely believed to be a useful chemical substance for preventing dental caries. However, the mechanism underlying crystal perforation in the tooth enamel and the effect of fluoride on hard tissues are unclear. To clarify the mechanism of the biological action of fluoride in the mineralization process, we examined the hard tissues of rats having received water containing a relatively low fluoride level. Electron microscopy revealed that fluoride ions could interrupt the crystal nucleation process, resulting in crystal perforation in the developing tooth enamel and the presence of amorphous minerals in bone crystals. Furthermore, the results of enzymatic analyses indicated that fluoride directly interfered with the synthesis of carbonic anhydrase by the enamel-forming cells, rather than being directly involved in the crystal formation. From the results, we would like to provide a possible mechanism of crystal perforation in the enamel induced by fluoride intake. Also, we would like to suggest that regardless of its amount, fluoride intake has harmful effects on both tooth and bone formation. PMID- 17419551 TI - Quantitative analysis of the calcium and phosphorus content of developing and permanent human teeth. AB - It was the aim of this study to investigate the distribution of Ca, P and C in predentin, dentin and enamel in human tooth buds and permanent teeth by EDX element analysis. The mandible of a 16-week-old human fetus containing eight mineralizing tooth buds and three human permanent molars were fixed in formaldehyde and embedded in Technovit 9100. Serial sections of 80 microm thickness of the mandible were cut in the frontal-dorsal direction, and polarized light micrographs were taken of these sections. The permanent teeth were cut in mesio-distal direction. The sections were investigated with scanning electron microscopy and EDX element analysis with a Philips XL 30 FEG scanning microscope and an EDAX energy-dispersive X-ray system using spot measurements, EDX line scans and element mapping. Quantitative measurements were made in predentin, mineralizing dentin adjacent to predentin, mature dentin, mineralizing enamel and young enamel of developing teeth and mature enamel of permanent teeth. In developing teeth the Ca and P content increased rapidly from outer predentin towards mineralizing dentin. In enamel prisms of developing teeth the Ca and P content increased linearly from the surface towards the enamel-dentin junction. In permanent teeth only a small layer of predentin was found. The Ca and P content in enamel and circumpulpal dentin of permanent teeth was higher than in developing teeth. The Ca/P ratio differed between predentin and dentin areas reflecting different calcium phosphate compositions, but it was the same in mineralizing and young enamel. The differences in the distribution of Ca and P reflect different mineralizing patterns of the enamel and dentin matrices. PMID- 17419552 TI - Preureteric inferior vena cava with possible rudiment of the proper inferior vena cava. AB - A right single preureteric inferior vena cava (IVC) was found in the cadaver of a 77-year-old Japanese male during a student dissection course at Kumamoto University School of Medicine in 2003. The ureter emerged from the lower end of the hilum of the right kidney at the second lumbar vertebral level. It ran inferomedially to pass behind the IVC, and turned inferolaterally to cross the vein superficially at the level of the third to the fourth lumbar vertebrae. Then, the ureter was situated to the right of the IVC, and descended ordinarily. The second lumbar vein of each side united bilaterally, as did the third lumbar veins. The common stem of the second lumbar veins drained into the left side of the IVC posterolaterally at the level of the second intervertebral disc, and the third common stem opened into the left border of the IVC at the fourth lumbar vertebral level. The ureter hooked around the IVC between the openings of those common stems. There was a small continuation (0.2 mm in diameter) between the left second lumbar and the right third lumbar veins along the vertebral column slightly right of the midline. It passed superficial to the right third lumbar artery, as did the IVC. The right testicular vein opened into the IVC at the level of the lower end of the third lumbar vertebral body. Generally, the level of the opening of the gonadal vein corresponds to the level of the caudal end of the remaining subcardinal vein, but it is lower than usual in this case. Furthermore, the segment from the confluence of the common iliac veins to the common trunk of the third lumbar veins, and to the small continuation can be regarded as the proper IVC, and the part where the ureter hooks around it may have derived from the anastomosis between the common trunk of the third lumbar veins and the subcardinal vein. PMID- 17419553 TI - A double aortic arch with a deformed trachea accompanied by a subaortic left innominate vein. AB - During our dissection of a Japanese elderly female cadaver, a double aortic arch with a deformed trachea was found in the cadaver. The ascending aorta was bifurcated to form the left (anterior) and right (posterior) aortic arches. Encircling and compressing the trachea and esophagus, they confluenced into the descending aorta. We concluded that it was a case of the double aortic arch forming a vascular ring. In the vascular ring the trachea was deformed. In addition, the left innominate vein coursed under the aortic arches (subaortic left innominate vein, SLIV) and crossed the mediastinum posterior to the ascending aorta and anterior to the trachea. PMID- 17419554 TI - Morphofunctional changes of orofacial muscles in patients with unilateral or bilateral cleft lip, alveolus and palate. AB - The facial musculature is part of a biocybernetic feedback system within the stomatognathic system, the continuity of which is disturbed by clefts of the lip, jaw, and palate (cheilognathopalatoschisis). This results in topographic, fine structural and functional alterations of the facial musculature. In two heads with unilateral clefts and two heads with bilateral clefts, the facial musculature was dissected and the M. orbicularis oris macroscopically and topographically examined. We found changes in the attachment of the M. orbicularis oris. The modiolus, which is coresponsible for the proper topographic relation of the M. obicularis oris to the other facial muscles, was unchanged. Further, we examined the vascular system of the muscle, which likewise adapts to the altered situation. Lip/jaw/palatal clefts result in anatomically functional and fine-structural alterations of the M. orbicularis oris, while the rest of the facial musculature remains unchanged. PMID- 17419562 TI - [Debridement in practice]. PMID- 17419563 TI - [Art therapy and the esthetics of care, a "phenomenal" union]. PMID- 17419564 TI - [Study on the administration of goserelin. Methodology and results]. PMID- 17419565 TI - [Prostate cancer and care. When male intimacy is put to the test...]. PMID- 17419566 TI - [Prostate cancer epidemiology]. PMID- 17419567 TI - [Prostate cancer physiopathology, diagnosis and therapy]. PMID- 17419568 TI - [Management of the patient after a radical prostatectomy]. PMID- 17419569 TI - [Conformal radiotherapy of prostate cancers]. PMID- 17419570 TI - [Brachytherapy of prostate cancers]. PMID- 17419571 TI - [Treatment by focalized high intensity ultrasound and nursing care]. PMID- 17419572 TI - [Urologic rehabilitation after prostate surgery]. PMID- 17419573 TI - [Nurse's role in the patient care]. PMID- 17419574 TI - [Prostate cancer and sexuality]. PMID- 17419575 TI - [Teaching the patient about intra-cavernous injections to prevent erectile dysfunction]. PMID- 17419576 TI - [Psychological management in medical oncology]. PMID- 17419577 TI - [Helping the patient in everyday life, the community nurse is a partner in shared care]. PMID- 17419578 TI - [Management of intimacy and professional care]. PMID- 17419579 TI - [Prostate cancer and care. Bibliography]. PMID- 17419580 TI - [What to remember about prevention and screening]. PMID- 17419581 TI - [2/10 Epidemiology of nosocomial infections]. PMID- 17419582 TI - [2/3 COX-2 inhibitors]. PMID- 17419583 TI - Ultrasound-mediated gene delivery: influence of contrast agent on transfection. AB - MAT B III cells were insonified at ultrasound frequencies of 1.15 and 2.25 MHz in the presence of different ultrasound contrast agents (UCA) and a plasmid encoding for the green fluorescent protein. The transfection efficiency was assessed by flow cytometry, while contrast agent destruction by ultrasound was evaluated using optical and scanning electronic microscopies. It was found that the gas and shell properties of the UCA have an important influence on cell transfection. A good correlation was observed between bubble destruction and transfection rate, and it was demonstrated, for the first time, that hard-shelled contrast agents (gas microcapsules) are promising candidates for ultrasound-mediated gene delivery. PMID- 17419584 TI - Characterization of photo-cross-linked oligo[poly(ethylene glycol) fumarate] hydrogels for cartilage tissue engineering. AB - Photo-cross-linkable oligo[poly(ethylene glycol) fumarate] (OPF) hydrogels have been developed for use in tissue engineering applications. We demonstrated that compressive modulus of these hydrogels increased with increasing polymer concentration, and hydrogels with different mechanical properties were formed by altering the ratio of cross-linker/polymer in precursor solution. Conversely, swelling of hydrogels decreased with increasing polymer concentration and cross linker/polymer ratio. These hydrogels are degradable and degradation rates vary with the change in cross-linking level. Chondrocyte attachment was quantified as a method for evaluating adhesion of cells to the hydrogels. These data revealed that cross-linking density affects cell behavior on the hydrogel surfaces. Cell attachment was greater on the samples with increased cross-linking density. Chondrocytes on these samples exhibited spread morphology with distinct actin stress fibers, whereas they maintained their rounded morphology on the samples with lower cross-linking density. Moreover, chondrocytes were photoencapsulated within various hydrogel networks. Our results revealed that cells encapsulated within 2-mm thick OPF hydrogel disks remained viable throughout the 3-week culture period, with no difference in viability across the thickness of hydrogels. Photoencapsulated chondrocytes expressed the mRNA of type II collagen and produced cartilaginous matrix within the hydrogel constructs after three weeks. These findings suggest that photo-cross-linkable OPF hydrogels may be useful for cartilage tissue engineering and cell delivery applications. PMID- 17419585 TI - Using small-angle neutron scattering to study the solution conformation of N-(2 hydroxypropyl)methacrylamide copolymer-doxorubicin conjugates. AB - Our past research developed two N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-doxorubicin (Dox) conjugates that became the first synthetic polymer anticancer conjugates to be evaluated clinically. The first, FCE28068, contained Dox bound to the polymeric carrier via a tetrapeptidic linker (glycine phenylalanine-leucine-glycine (GFLG)) (Mw approximately 30,000 g/mol; approximately 8 wt % drug), and the second, FCE28069, contained additionally galactosamine (Gal) (Mw approximately 30,000 g/mol; approximately 7.5 wt % Dox) again bound by a GFLG linker. Galactosamine was included to promote hepatocyte/hepatoma targeting via the asialoglycoprotein receptor. Both conjugates showed antitumor activity and were clinically less toxic than free Dox (2-5 fold). However, despite their similar chemical characteristics, the conjugates displayed a significantly different maximum-tolerated dose (MTD) in patients. The aim of this study, therefore, was to use small-angle neutron scattering (SANS) to explore the solution behavior of a small library of HPMA polymer conjugates including FCE28068, FCE28069, and their pharmaceutical formulations, plus as reference compounds HPMA copolymer-GFLG conjugates containing aminopropanol (Ap) or galactosamine (Gal) alone (i.e., without Dox). The SANS data obtained showed that HPMA copolymer-GFLG-Ap conjugates (containing 5 and 10 mol % side chains) showed evidence of polymer aggregation, however, no indication of aggregation was observed for FCE28068 and FCE28069 over the concentration range studied (2.5-50 mg/mL). Clear differences in the scattering behavior for the two conjugates were observed at equivalent concentration. Data were best fitted by a model for polydisperse Gaussian coils, and the HPMA copolymer-Dox conjugate with Gal (FCE28069) exhibited a larger radius of gyration (Rg) (by approximately 2.5 nm) compared to FCE28068. In conclusion, we have shown that SANS will be a valuable tool to elucidate conformation-performance relationships for polymer-drug conjugates. PMID- 17419586 TI - Interaction of chitosan with cell membrane models at the air-water interface. AB - In this paper we employed phospholipid Langmuir monolayers as membrane models to probe interactions with chitosan. Using a combination of surface pressure--area and surface potential--area isotherms and rheological measurements with the pendent drop technique, we observed that chitosan interacts with phospholipid molecules at the air-water interface. We propose a model in which chitosan interacts with the phospholipids mainly through electrostatic interactions, but also including H-bonding and hydrophobic forces, depending on the phospholipid packing density. At large areas per molecule, chitosan in the subphase adsorbs onto the monolayer, expanding it. At small areas per molecule, chitosan is located in the subsurface. Indeed, a mixed chitosan-phospholipid monolayer can be transferred onto solid supports, even at high surface pressures. The effects of chitosan on the viscoelastic properties of phospholipid monolayers may be taken as evidence for the ability of chitosan to disrupt cell membranes. PMID- 17419587 TI - Electrochemical NO2 sensor using a NiFe1.9Al0.1O4 oxide spinel electrode. AB - A novel solid-state electrochemical sensor using (Sc2O3)0.08(ZrO2)0.92 (ScSZ) electrolyte solid and a NiFe1.9Al0.1O4 oxide spinel electrode was tested for the detection of NO2 at temperatures greater than 700 degrees C for automobile applications. The sensor was found to respond rapidly, reproducibly, and selectively to NO2 at 703 and 740 degrees C. The response time of the sensor was approximately 8 s, and the recovery time was 10 s at both 703 and 741 degrees C. The response of the sensor was highly reproducible to the change in concentration of NO2 and also showed negligible cross-sensitivity to potentially interfering gases such as O2, CO, and CH4 in the gas stream. PMID- 17419588 TI - Silica colloidal crystals for enhanced fluorescence detection in microarrays. AB - Silica colloidal crystals were investigated for their potential as high surface area materials to enhance sensitivity over planar surfaces for microarrays using fluorescence detection. A relation was derived showing how crystal thickness and transmission, as well as colloid size, combine to determine the optically accessible surface area for enhancing sensitivity. Experimentally, crystals of 250-nm colloids were prepared with thicknesses determined by SEM to be 1.6, 4.2, and 11.0 microm. The material was sintered at 1000 degrees C to make it durable without affecting the crystalline structure, as confirmed by SEM. UV/visible spectrometry showed the depth of penetration (1/e) to be 8.4 microm at 488 nm for these materials. Fluorescein-labeled streptavidin and biotin were used as a model ligand-receptor pair. For the fluorescence measurements, biotin was covalently bonded to the silica surfaces, and the fluorescence was detected from the captured streptavidin-fluorescein. The observed fluorescence enhancement agreed well with the theory developed here. Compared to a planar surface, the colloidal crystal of 11.0 microm in thickness enhanced the fluorescence by nearly a factor of 80, with only a 0.3% increase in fluorescence background. PMID- 17419589 TI - Deletion of the P5abc peripheral element accelerates early and late folding steps of the Tetrahymena group I ribozyme. AB - The P5abc peripheral element stabilizes the Tetrahymena group I ribozyme and enhances its catalytic activity. Despite its beneficial effects on the native structure, prior studies have shown that early formation of P5abc structure during folding can slow later folding steps. Here we use a P5abc deletion variant E(deltaP5abc) to systematically probe the role of P5abc throughout tertiary folding. Time-resolved hydroxyl radical footprinting shows that E(deltaP5abc) forms its earliest stable tertiary structure on the millisecond time scale, approximately 5-fold faster than the wild-type ribozyme, and stable structure spreads throughout E(deltaP5abc) in seconds. Nevertheless, activity measurements show that the earliest detectable formation of native E(deltaP5abc) ribozyme is much slower (approximately 0.6 min(-1)), in a manner similar to that of the wild type. Also similar, only a small fraction of E(deltaP5abc) attains the native state on this time scale under standard conditions at 25 degrees C, whereas the remainder misfolds; footprinting experiments show that the misfolded conformer shares structural features with the long-lived misfolded conformer of the wild type ribozyme. Thus, P5abc does not have a large overall effect on the rate limiting step(s) along this pathway. However, once misfolded, E(deltaP5abc) refolds to the native state 80-fold faster than the wild-type ribozyme and is less accelerated by urea, indicating that P5abc stabilizes the misfolded structure relative to the less-ordered transition state for refolding. Together, the results suggest that, under these conditions, even the earliest tertiary folding intermediates of the wild-type ribozyme represent misfolded species and that P5abc is principally a liability during the tertiary folding process. PMID- 17419590 TI - A unified kinetic mechanism applicable to multiple DNA polymerases. AB - After extensive studies spanning over half a century, there is little consensus on the kinetic mechanism of DNA polymerases. Using stopped-flow fluorescence assays for mammalian DNA polymerase beta (Pol beta), we have previously identified a fast fluorescence transition corresponding to conformational closing, and a slow fluorescence transition matching the rate of single nucleotide incorporation. Here, by varying pH and buffer viscosity, we have decoupled the rate of single-nucleotide incorporation from the rate of the slow fluorescence transition, thus confirming our previous hypothesis that this transition represents a conformational event after chemistry, likely subdomain reopening. Analysis of an R258A mutant indicates that rotation of the Arg258 side chain is not rate-limiting in the overall kinetic pathway of Pol beta, yet is kinetically significant in subdomain reopening. We have extended our kinetic analyses to a high-fidelity polymerase, Klenow fragment (KF), and a low-fidelity polymerase, African swine fever virus DNA polymerase X (Pol X), and showed that they follow the same kinetic mechanism as Pol beta, while differing in relative rates of single-nucleotide incorporation and the putative conformational reopening. Our data suggest that the kinetic mechanism of Pol beta is not an exception among polymerases, and furthermore, its delineated kinetic mechanism lends itself as a platform for comparison of the kinetic properties of different DNA polymerases and their mutants. PMID- 17419592 TI - Experimental and theoretical study on the structure and formation mechanism of [C6H5Cu(m)]- (m = 1-3). AB - The important intermediate phenyl-copper metal complexes [C(6)H(5)Cu(m)]- (m = 1 3), which are produced from the reactions between copper metal clusters formed by laser ablation and the benzene molecules seeded in argon carrier gas, are studied by photoelectron spectroscopy(PES) and density functional theory (DFT). Their structures and bonding patterns are investigated, which results in the conclusion that C(6)H(5) groups bond perpendicularly on copper clusters through Cu-C sigma bond. The formation mechanism of these complexes has been studied at B3LYP//6 311G(d, p)/Lanl2dz level. Direct insertion reaction between [Cu(m)]- and C(6)H(6) yields intermediate complex [C(6)H(5)Cu(m)H]-, and then eliminates the H atom, or releases the H atom to other neutral Cu atoms or anionic Cu ions via H abstraction reaction. The first step is the rate-limiting step with C-H activation and cleavage, and H abstraction by neutral Cu atom is the most energetically favorable pathway for the final step. Moreover, the complex [C(6)H(5)Cu(2)]- is ascertained to be easier to be generated than [C(6)H(5)Cu(3)] and [C(6)H(5)Cu]-, which are in excellent agreement with the experimental results. PMID- 17419591 TI - Selenoglutathione: efficient oxidative protein folding by a diselenide. AB - Diselenide bonds are intrinsically more stable than disulfide bonds. To examine how this stability difference affects reactivity, we synthesized selenoglutathione (GSeSeG), an analogue of the oxidized form of the tripeptide glutathione that contains a diselenide bond in place of the natural disulfide. The reduction potential of this diselenide bond was determined to be -407 +/- 9 mV, a value which is 151 mV lower than that of the disulfide bond in glutathione (GSSG). Thus, the diselenide bond of GSeSeG is 7 kcal/mol more stable than the disulfide bond of GSSG. Nonetheless, we found that GSeSeG can be used to oxidize cysteine residues in unfolded proteins, a process that is driven by the gain in protein conformational stability upon folding. Indeed, the folding of both ribonuclease A (RNase A) and bovine pancreatic trypsin inhibitor (BPTI) proceeded efficiently using GSeSeG as an oxidant, in the former case with a 2-fold rate increase relative to GSSG and in the latter case accelerating conversion of a stable folding intermediate to the native state. In addition, GSeSeG can also oxidize the common biological cofactor NADPH and is a good substrate for the NADPH-dependent enzyme glutathione reductase (kcat = 69 +/- 2 s-1, Km = 54 +/- 7 microM), suggesting that diselenides can efficiently interact with the cellular redox machinery. Surprisingly, the greater thermodynamic stability of diselenide bonds relative to disulfide bonds is not matched by a corresponding decrease in reactivity. PMID- 17419593 TI - Vibrational assignment of the Raman active modes of 1,10-phenanthroline-5,6-dione using DFT calculations. AB - A complete assignment of the Raman active modes of 1,10-phenanthroline-5,6-dione in the 100-4000 cm(-1) spectral region is reported. Intense well resolved spectra of solid phendione with high S/N are reported. Assignment of the normal modes with appropriate symmetry representation symbols was achieved by employing density functional theory calculations. Our calculations were modeled on results previously reported for phenanthroline. Results of the B3LYP calculations were consistent and established that phendione possess sixty fundamentals. PMID- 17419594 TI - Improved determination of structural changes of 2-pyridone-(H2O)1 upon electronic excitation. AB - The change of the 2-pyridone-water cluster (2PYH(2)O) structure upon electronic excitation is determined by a Franck-Condon analysis of the intensities in the fluorescence emission spectra obtained via excitation of three different vibronic bands as well as a structural fit based on the rotational constants of eight isotopomers that have been reported by Held and Pratt (J. Am. Chem. Soc., 1993, 115, 9708]. A total of 93 emission band intensities were fit, together with the changes of rotational constants of 8 isotopomers. The geometry change upon electronic excitation to the pipi* state can be described by a strong and unsymmetrical elongation of the hydrogen bonds, a contraction of the OH bond involved in the cyclic cluster arrangement, and an unsymmetrical ring deformation. The resulting geometry changes are interpreted on the basis of ab initio calculations. PMID- 17419595 TI - Computational study of the reaction between biogenic stabilized Criegee intermediates and sulfuric acid. AB - We have postulated a mechanism for the reaction of sulfuric acid with stabilized Criegee intermediates (sCIs). We have computed Gibbs free energies for the reaction of sulfuric acid with two biogenic sCIs and three smaller model species. We have also calculated Gibbs free energies for two competing sink reactions. Due to the large size of the biogenic sCIs, the computations have been performed at the relatively modest B3LYP/6-31G(d,p) and B3LYP/6-311+G(2d,p) levels. However, single-point RI-CC2/def2-QZVPP calculations for the (CH3)(2)COO model species are in good agreement with the B3LYP results. The reaction is found to be strongly exothermic for all studied species. Activation barrier calculations on the (CH3)(2)COO model species further indicate that the reaction with sulfuric acid may proceed significantly faster than the sink reaction with water. If the same applies to the biogenic sCIs, the proposed reactions could account for some part of the organically assisted new particle formation events observed in the atmosphere. PMID- 17419596 TI - Ab initio characterization of (CH3IO3) isomers and the CH3O2 + IO reaction pathways. AB - The geometries, harmonic vibrational frequencies, relative energetics, and enthalpies of formation of (CH(3)IO(3)) isomers and the reaction CH(3)O(2) + IO have been investigated using quantum mechanical methods. Optimization has been performed at the MP2 level of theory, using all electron and effective core potential, ECP, computational techniques. The relative energetics has been studied by single-point calculations at the CCSD(T) level. Methyl iodate, CH(3)OIO(2), is found to be the lowest-energy isomer showing particular stabilization. The two nascent association minima, CH(3)OOOI and CH(3)OOIO, show similar stabilities, and they are considerably higher located than CH(3)OIO(2). Interisomerization barriers have been determined, along with the transition states involved in various pathways of the reaction CH(3)O(2) + IO. PMID- 17419597 TI - Nonlinear vibrational spectroscopic studies of the adsorption and speciation of nitric acid at the vapor/acid solution interface. AB - Nitric acid plays an important role in the heterogeneous chemistry of the atmosphere. Reactions involving HNO(3) at aqueous interfaces in the stratosphere and troposphere depend on the state of nitric acid at these surfaces. The vapor/liquid interface of HNO(3)-H2O binary solutions and HNO(3)-H(2)SO(4)-H2O ternary solutions are examined here using vibrational sum frequency spectroscopy (VSFS). Spectra of the NO2 group at different HNO(3) mole fractions and under different polarization combinations are used to develop a detailed picture of these atmospherically important systems. Consistent with surface tension and spectroscopic measurements from other laboratories, molecular nitric acid is identified at the surface of concentrated solutions. However, the data here reveal the adsorption of two different hydrogen-bonded species of undissociated HNO(3) in the interfacial region that differ in their degree of solvation of the nitro group. The adsorption of these undissociated nitric acid species is shown to be sensitive to the H2O:HNO(3) ratio as well as to the concentration of sulfuric acid. PMID- 17419598 TI - Linear solvation energy parameters for model tropospheric aerosol surfaces. AB - Excited-state absorption spectra for several coumarin derivatives adsorbed to aerosol particles provide linear solvation energy (LSE) relationships for the aerosol surfaces. This study focuses on NaCl and (NH4)2SO(4) particles as models for tropospheric aerosol. We investigate several others, including NH(4)Cl, NaBr, KI, Na(2)SO(4), NaNO(3), Al(2)O3, and CaCO(3), to establish trends and understand the factors that control polarity for surfaces. The Kamlet-Taft dipolarity/polarizability parameter, pi*, for these particles ranges from 0.73 to 1.69. The values are high compared to most homogeneous molecular solvents and are attributable to ion-dipole forces, especially at defect sites. We also find that the smaller values of pi* (1.01 for (NH4)2SO(4) and 0.73 for NH(4)Cl) correlate with appreciable hydrogen bond donor acidity in the surface (alpha = 0.23 and 1.06, respectively). Strong hydrogen bonds with the surface lead to a drop in overall polarity either by making interaction with very polar defect sites less likely or orienting the probe molecule away from the surface. Adsorbed water layers mainly alter the alpha value of the surface, but can have indirect effects on pi* by changing the interaction of the adsorbed molecule with the surface. PMID- 17419599 TI - Excited-state distortions of cyclometalated Ir(III) complexes determined from the vibronic structure in luminescence spectra. AB - The luminescence spectra of [(tpy)(2)Ir(CN-t-Bu)2](CF(3)SO(3)) in methylcyclohexane glass and frozen n-nonane at 15 K reveal well-resolved vibronic fine structure. The vibronic peaks are assigned by comparison with the vibrational frequencies obtained from Raman and IR spectra and those obtained using DFT electronic structure calculations. The magnitudes of the distortions along the normal coordinates are calculated by fitting the emission spectra using the time-dependent theory of spectroscopy. Broadening effects and the MIME frequency observed at room temperature are interpreted. The most highly distorted normal modes involve atomic motions on the tpy ligand, consistent with the metal to ligand/ligand centered assignment of the electronic transition. PMID- 17419600 TI - Vacuum-ultraviolet photoionization studies of the microhydration of DNA bases (guanine, cytosine, adenine, and thymine). AB - In this work, we report on a photoionization study of the microhydration of the four DNA bases. Gas-phase clusters of water with DNA bases [guanine (G), cytosine (C), adenine (A), and thymine (T)] are generated via thermal vaporization of the bases and expansion of the resultant vapor in a continuous supersonic jet expansion of water seeded in Ar. The resulting clusters are investigated by single-photon ionization with tunable vacuum-ultraviolet synchrotron radiation and mass analyzed using reflectron mass spectrometry. Photoionization efficiency (PIE) curves are recorded for the DNA bases and the following water (W) clusters: G, GWn (n = 1-3); C, CWn (n = 1-3); A, AWn (n = 1,2); and T, TWn (n = 1-3). Appearance energies (AE) are derived from the onset of these PIE curves (all energies in eV): G (8.1 +/- 0.1), GW (8.0 +/- 0.1), GW2 (8.0 +/- 0.1), and GW3 (8.0); C (8.65 +/- 0.05), CW (8.45 +/- 0.05), CW2 (8.4 +/- 0.1), and CW3 (8.3 +/- 0.1); A (8.30 +/- 0.05), AW (8.20 +/- 0.05), and AW2 (8.1 +/- 0.1); T (8.90 +/- 0.05); and TW (8.75 +/- 0.05), TW2 (8.6 +/- 0.1), and TW3 (8.6 +/- 0.1). The AEs of the DNA bases decrease slightly with the addition of water molecules (up to three) but do not converge to values found for photoinduced electron removal from DNA bases in solution. PMID- 17419601 TI - Axial halogen ligand effect on photophysics and optical power limiting of some indium naphthalocyanines. AB - Three axially substituted complexes, 2,3-octa(3,5-di-tert-butylphenoxy)-2,3 naphthalocyaninato indium chloride (1a), 2,3-octa(3,5-di-tert-butylphenoxy)-2,3 naphthalocyaninato indium bromide (1b), and 2,3-octa(3,5-di-tert-butylphenoxy) 2,3-naphthalocyaninato indium iodide (1c) have been synthesized and their photophysical properties have been investigated. Optical power limiting of nanosecond (ns) and picosecond (ps) laser pulses at 532 nm using these complexes has been demonstrated. All complexes display strong Q(0,0) absorption and measurable emission in the near-infrared region and exhibit strong excited-state absorption in the range of 470-700 nm upon ns laser excitation. The different axial ligands show negligible effect on the linear absorption, emission, and transient difference absorption spectra. However, the excited-state lifetime, triplet excited-state quantum yield, and efficiency to generate singlet oxygen are affected significantly by the heavier axial ligand. Brominated and iodinated complexes 1b and 1c show higher triplet excited-state quantum yield, while chlorinated complex 1a has longer excited-state lifetime and is more efficient in generating singlet oxygen. The iodinated complex 1c displayed the best optical limiting due to the higher ratio of excited-state absorption cross section to ground state absorption cross section (sigma(eff)/sigma(0)). PMID- 17419602 TI - A swift all-atom energy-based computational protocol to predict DNA-ligand binding affinity and DeltaTm. AB - A hybrid molecular mechanics-statistical mechanics-solvent accessibility-based computational protocol is developed to calculate DNA-ligand binding affinity without any database training and is validated on 50 DNA-ligand complexes. The calculated binding energies yield high correlation coefficients of 0.95 (R2 = 0.90) and 0.96 (R2 = 0.93) in linear plots against experimental binding free energies (DeltaGo) and DeltaTm, respectively. The protocol is translated into a swift, web-enabled, freely accessible computational tool, http://www.scfbio iitd.res.in/preddicta, for DeltaGo and DeltaTm prediction for DNA-ligand complexes to aid and expedite rational drug design attempts. PMID- 17419603 TI - Design and synthesis of a potent histone deacetylase inhibitor. AB - Histone deacetylase (HDAC) inhibitors have potential for cancer therapy. An HDAC inhibitor based on a cyclic peptide mimic of known structure, linked by an aliphatic chain to a hydroxamic acid, was designed and synthesized. The chimeric compound showed potent competitive inhibition of nuclear HDACs, with an IC50 value of 46 nM and a Ki value of 13.7 nM. The designed inhibitor showed 4-fold selectivity for HDAC1 (57 nM) over HDAC8 (231 nM). PMID- 17419604 TI - In vitro optimization of non-small cell lung cancer activity with troxacitabine, L-1,3-dioxolane-cytidine, prodrugs. AB - l-1,3-Dioxolane-cytidine, a potent anticancer agent against leukemia, has limited efficacy against solid tumors, perhaps due to its hydrophilicity. Herein, a library of prodrugs were synthesized to optimize in vitro antitumor activity against non-small cell lung cancer. N4-Substituted fatty acid amide prodrugs of 10-16 carbon chain length demonstrated significantly improved antitumor activity over l-1,3-dioxolane-cytidine. These in vitro results suggest that the in vivo therapeutic efficacy of l-1,3-dioxolane-cytidine against solid tumors may be improved with prodrug strategies. PMID- 17419605 TI - Radiolabeled phenethylguanidines: novel imaging agents for cardiac sympathetic neurons and adrenergic tumors. AB - The norepinephrine transporter (NET) substrates [123I]-m-iodobenzylguanidine (MIBG) and [11C]-m-hydroxyephedrine (HED) are used as markers of cardiac sympathetic neurons and adrenergic tumors (pheochromocytoma, neuroblastoma). However, their rapid NET transport rates limit their ability to provide accurate measurements of cardiac nerve density. [11C]Phenethylguanidine ([11C]1a) and 12 analogues ([11C]1b-m) were synthesized and evaluated as radiotracers with improved kinetics for quantifying cardiac nerve density. In isolated rat hearts, neuronal uptake rates of [11C]1a-m ranged from 0.24 to 1.96 mL min-1 (g wet wt) 1, and six compounds had extremely long neuronal retention times (clearance T1/2 > 20 h) due to efficient vesicular storage. Positron emission tomography (PET) studies in nonhuman primates with [11C]1e, N-[11C]guanyl-m-octopamine, which has a slow NET transport rate, showed improved myocardial kinetics compared to HED. Compound [11C]1c, [11C]-p-hydroxyphenethylguanidine, which has a rapid NET transport rate, avidly accumulated into rat pheochromocytoma xenograft tumors in mice. These encouraging findings demonstrate that radiolabeled phenethylguanidines deserve further investigation as radiotracers of cardiac sympathetic innervation and adrenergic tumors. PMID- 17419606 TI - Development of ruthenium antitumor drugs that overcome multidrug resistance mechanisms. AB - Organometallic ruthenium(II) complexes of the general formula [Ru(eta6-p cymene)Cl2(L)] and [Ru(eta6-p-cymene)Cl(L)2][BPh4] with modified phenoxazine- and anthracene-based multidrug resistance (MDR) modulator ligands (L) have been synthesized, spectroscopically characterized, and evaluated in vitro for their cytotoxic and MDR reverting properties in comparison with the free ligands. For an anthracene-based ligand, coordination to a ruthenium(II) arene fragment led to significant improvement of cytotoxicity as well as Pgp inhibition activity. A similar, but weaker effect was also observed when using a benzimidazole phenoxazine derivative as Pgp inhibitor. The most active compound in terms of both Pgp inhibition and cytotoxicity is [Ru(eta6-p-cymene)Cl2(L)], where L is an anthracene-based ligand. Studies show that it induces cell death via inhibition of DNA synthesis. Moreover, because the complex is fluorescent, its uptake in cells was studied, and relative to the free anthracene-based ligand, uptake of the complex is accelerated and accumulation of the complex in the cell nucleus is observed. PMID- 17419607 TI - Synthesis and biological evaluation of 2- and 3-aminobenzo[b]thiophene derivatives as antimitotic agents and inhibitors of tubulin polymerization. AB - Two new series of inhibitors of tubulin polymerization based on the 2-amino-3 (3,4,5-trimethoxybenzoyl)benzo[b]thiophene molecular skeleton and its 3-amino positional isomer were synthesized and evaluated for antiproliferative activity, inhibition of tubulin polymerization, and cell cycle effects. Although many more 3-amino derivatives have been synthesized so far, the most promising compound in this series was 2-amino-6-methyl-3-(3,4,5-trimethoxybenzoyl)benzo[b]thiophene, which inhibits cancer cell growth at subnanomolar concentrations and interacts strongly with tubulin by binding to the colchicine site. PMID- 17419608 TI - Nuclear phospholipase C gamma: punctate distribution and association with the promyelocytic leukemia protein. AB - The marriage between transducers of cell stress stimuli and their nuclear targets is likely to be achieved in part by some spatial-temporal compartmentalization of the relevant effectors. A candidate compartment for these events is the promyelocytic leukemia nuclear domain (PML-ND), within which are found numerous effectors of damage recognition, repair, and cell death. We predicted that the identification of PML-ND cargo proteins would clarify those biochemical pathways that straddle the recognition of cellular damage and cell fate. We now use mass spectrometry of peptides eluted from PML coprecipitates to demonstrate that the gamma 1 (gamma1) isoform of PLC associates with nuclear PML. Though thought to act primarily in the cytoplasm, we use biochemical fractionation combined with immunocytochemistry to verify the nuclear expression of PLC-gamma1 and its interaction with PML. These are the first data to show an interaction between endogenous levels of a phosphoinositide metabolizing protein and the biophysically labile PML-ND by mass spectrometry and add weight to the view that PML-NDs may act as tumor suppressors by sequestering mitogenic effectors. PMID- 17419609 TI - Terahertz time-domain spectroscopy of imidazolium ionic liquids. AB - We have measured the terahertz (THz) complex dielectric spectra of imidazolium ionic liquids by THz time-domain spectroscopy (THz-TDS) in the frequency range from 5 (0.15 THz) to 140 cm(-1) (4.2 THz). The ionic liquids investigated are 1 ethyl-3-methylimidazolium (EMIm+)/trifluoromethanesulfonate (TfO-), EMIm+/tetrafluoroborate (BF(4)-), 1-butyl-3-methylimidazolium (BMIm+)/TfO-, and BMIm+/BF(4)-. The dielectric values of the ionic liquids in the THz region are similar to those of short-chain alcohols. The THz dielectric values are related to subpicosecond-to-picosecond dynamics. The same trend has been observed in the empirical polarity ET(30) although it is related to the static characteristics of polarity and hydrogen bonding ability. A difference between the two types of liquids is observed in the THz dielectric spectral shapes: the ionic liquids show structured lineshapes but short-chain alcohols show much less structured ones. The structured lineshapes of the ionic liquids reflect the low-frequency motions of interion and/or intramolecular vibrations. When the ionic liquids composed of the different imidazolium cations contain the same anions as counterions, their density-normalized THz dielectric spectra above 20 cm(-1) bear strong resemblance to each other in shape and magnitude. It shows clearly that the THz spectra do not originate from the intramolecular vibrations of the imodazolium cations. All of the intramolecular vibrations of the anions are located above 140 cm(-1) except the CF3-SO3 torsion of TfO-, the band of which alone cannot explain the broad THz dielectric spectra of the ionic liquids. Therefore, we conclude that the interion vibrations rather than the intramolecular vibrations dominantly contribute to the THz dielectric spectra. The results strongly indicate that even in the liquid phase the ionic liquids have local structures similar to their solid-phase structures. PMID- 17419610 TI - A bias-exchange approach to protein folding. AB - By suitably extending a recent approach [Bussi, G.; et al. J. Am. Chem. Soc. 2006, 128, 13435] we introduce a powerful methodology that allows the parallel reconstruction of the free energy of a system in a virtually unlimited number of variables. Multiple metadynamics simulations of the same system at the same temperature are performed, biasing each replica with a time-dependent potential constructed in a different set of collective variables. Exchanges between the bias potentials in the different variables are periodically allowed according to a replica exchange scheme. Due to the efficaciously multidimensional nature of the bias the method allows exploring complex free energy landscapes with high efficiency. The usefulness of the method is demonstrated by performing an atomistic simulation in explicit solvent of the folding of a Triptophane cage miniprotein. It is shown that the folding free energy landscape can be fully characterized starting from an extended conformation with use of only 40 ns of simulation on 8 replicas. PMID- 17419611 TI - Structure and hydration of L-proline in aqueous solutions. AB - The structure and hydration of L-proline in aqueous solution have been investigated using a combination of neutron diffraction with isotopic substitution, empirical potential structure refinement modeling, and small-angle neutron scattering at three concentrations, 1:10, 1:15, and 1:20 proline/water mole ratios. In each solution the carboxylate oxygen atoms from proline accept less than two hydrogen bonds from the surrounding water solvent and the amine hydrogen atoms donate less than one hydrogen bond to the surrounding water molecules. The solute-solute radial distribution functions indicate relatively weak interactions between proline molecules, and significant clustering or aggregation of proline is absent at all these concentrations. The spatial density distributions for the hydration of the COO- group in proline show a similar shape to that found previously in L-glutamic acid in aqueous solution but with a reduced coordination number. PMID- 17419612 TI - Infrared study of the effect of hydration on the amide I band and aggregation properties of helical peptides. AB - The amide I' band of a polypeptide is sensitive not only to its secondary structure content but also to its environment. In this study we show how degrees of hydration affect the underlying spectral features of the amide I' band of two alanine-based helical peptides. This is achieved by solubilizing these peptides in the water pool of sodium bis(2-ethylhexyl)sulfosuccinate reverse micelles with different water contents or w0 values. In agreement with several earlier studies, our results show that the amide I' band arising from a group of dehydrated helical amides is centered at approximately 1650 cm-1, whereas hydration shifts this frequency toward lower wavenumbers. More importantly, temperature-dependent infrared studies further show that these helical peptides undergo a thermally induced conformational transition in reverse micelles of low w0 values (e.g., w0=6), resulting in soluble peptide aggregates rich in antiparallel beta-sheets. Interestingly, however, increasing w0 or water content leads to an increase in the onset temperature at which such beta-aggregates begin to form. Therefore, these results provide strong evidence suggesting that dehydration facilitates aggregate formation and that removal of water imposes a free energy barrier to peptide association and aggregation, a feature that has been suggested in recent simulation studies focusing on the mechanism of beta-amyloid formation. PMID- 17419613 TI - Influence of the silane modifiers on the surface thermodynamic characteristics and dispersion of the silica into elastomer compounds. AB - A good dispersion of silica into elastomers, typically used in tire tread production, is obtained by grafting of the silica with multifunctional organosilanes. In this study, the influence of the chemical structure of a triethoxysilane (TES), octadecyltriethoxysilane (ODTES), and ODTES/bistriethoxysilylpropyltetrasulfane (TESPT) mixture was investigated by inverse gas chromatography (IGC) at infinite dilution. Thermodynamic results indicate a higher polarity of the silica surface modified with TES as compared to that of the unmodified silica due to new OH groups deriving from the hydrolysis of ethoxy groups of the silane; the long hydrocarbon substituent of the ODTES lies on the surface of silica and reduces the dispersive component of the silica surface tension. A comparison with silica modified with TESPT is discussed. An accurate morphological investigation by transmission electron microscopy (TEM) and automated image analysis (AIA) was carried out on aggregates of silica dispersed into a SBR compound loaded with 35 phr (per hundred rubber) of untreated and TESPT-treated silica. Morphological descriptors such as the projected area/perimeter ratio (A/P) and roundness (P2/4piA) provided direct and quantitative indications about the distribution of the filler into the rubber matrix. PMID- 17419614 TI - A simple route to synthesize scales of aligned single-crystalline SiC nanowires arrays with very small diameter and optical properties. AB - Scales of aligned single-crystalline SiC nanowires (SiCNWs) arrays with very small diameter were synthesized by a simple thermal evaporation of ZnS and carbon on silicon wafer. The as-received SiCNWs possess a uniform size distribution centered at approximately 8.0 nm, even with a minimum of approximately 3.0 nm. The highly oriented SiCNWs usually grew along [111] direction with a clean surface, very thin oxide shell, and small quantity of stacking faults. A crystalline tube-like SiC nanostructure is also obtained. The optical properties, including photoluminescence and Raman scattering spectra of the SiCNWs, were investigated, respectively. In the end, a growth model on basis of the experimental data is suggested. PMID- 17419615 TI - Square-planar N2S2NiII complexes with an extended pi-conjugated system. AB - The reaction of a mixture of 2-(1-naphthyl)benzothiazoline (HL1) and 2,6 diphenylbenzo[1,2-d:4,5-d']bisthiazoline (H3L2) with nickel(II) acetate tetrahydrate yielded three kinds of square-planar nickel(II) complexes: one nickel(II) complex with innocent ligands ([Ni(L1)2] (1c)) and two nickel(II) complexes with non-innocent ligands ([Ni(L1-L1)] (1a) and [Ni(L1-L2)] (1b)). The complex 1c has two bidentate-N,S ligands, which are formed via ring opening of HL1. On the other hand, the two complexes 1a and 1b contain a tetradentate-N2S2 ligand, which is created via ring opening of HL1 and H3L2, followed by bond formation between imino carbon atoms. Complexes 1a and 1b show very intense absorptions in the near-infrared (NIR) region, characteristic of square-planar complexes with non-innocent ligands. The third nickel(II) complex having a non innocent tetradentate-N2S2 ligand ([Ni(L2-L2)] (2)) was prepared from H3L2 and nickel(II) acetate tetrahydrate. The electronic spectrum of 2 exhibits a very intense absorption at 981 nm (epsilon = 3.6 x 10(4) M-1 cm-1), which is significantly red-shifted compared with those of 1a (837 nm and 4.4 x 10(4) M-1 cm-1) and 1b (885 nm and 4.5 x 10(4) M-1 cm-1), indicating the presence of an extended pi delocalization. The reaction of 2,6-bis(3,5-dichlorophenyl)benzo[1,2 d:4,5-d']bisthiazoline (H3L3) with nickel(II) acetate tetrahydrate also led to the formation of a nickel(II) complex with a non-innocent ligand ([Ni(L3-L3)] (3)). While complex 3 is analogous to 2, its electrical conductivity is much higher than that of 2. The molecular structures of 1b, 1c, 2, and 3 were determined by X-ray crystallography. PMID- 17419616 TI - Synthesis and characterization of vanadium(IV) complexes with cis-inositol in aqueous solution and in the solid-state. AB - The complex formation of vanadium(IV) with cis-inositol (ino) and the corresponding trimethyl ether 1,3,5-trideoxy-1,3,5-trimethoxy-cis-inositol (tmci) was studied in aqueous solution and in the solid-state. With increasing pH, the formation of [VO(H-2L)], [(VO)2L2H-5]-, [VO(H-3L)]- (L = ino) or [(VO)2L2H-6]2- (L = tmci), [V(H-3L)2]2-, and [VO(H-3L)(OH)2]3- was observed. For the vanadium(IV)/ino system, [(VO)2L2H-7]3- was observed as an additional dinuclear species. The formation constants of these complexes were determined by potentiometric titrations (25 degrees C, 0.1 M KCl). In addition, the vanadium(IV)/ino system was investigated by means of UV-vis spectrophotometric methods. EPR spectroscopy and cyclic voltammetry confirmed this complexation scheme. EPR measurements indicated the formation of three distinct isomers of the non-oxo complex [V(H-3ino)2]2- in weakly basic solution. This type of isomerism, which is not observed for the vanadium(IV)/tmci system, was assigned to the ability of ino to bind the vanadium(IV) center with three alkoxo groups having either a 1,3,5-triaxial or an 1,2,3-axial-equatorial-axial arrangement. The structures of [V(H-3ino)2][K2(ino)2].4H2O (1) and [Na6V(H-3ino)2](SO4)2.6H2O (2) were determined by single-crystal X-ray analysis. In both compounds, the coordination of each ino molecule to the vanadium(IV) center via three axial deprotonated oxygen donors was confirmed. The centrosymmetric structure of the coordination spheres corresponds to an almost regular octahedral geometry with a twist angle of 60 degrees. The crystal structure of the potassium complex 1 represents an unusual 1:1 packing of [V(H-3ino)2]2- dianions and [K2(ino)2]2+ dications, in which both K+ ions have a coordination number of nine and are bonded simultaneously to a 1,3,5-triaxial and an 1,2,3-axial-equatorial-axial site of ino. In 2, the [V(H-3ino)2]2- complexes are surrounded by six Na+ counterions that are bonded to the axial alkoxo oxygens and to the equatorial hydroxy oxygens of the cis-inositolato moieties. The six Na+ centers are further interlinked by bridging sulfate ions. According to EPR spectroscopy, the D3d symmetric structure of the [V(H-3ino)2]2- anion is retained in H2O, in dimethylformamide, and in a mixture of CHCl3/toluene 60:40 v/v. PMID- 17419617 TI - Vanadium(V) tartrato complexes: speciation in the H3O+(OH-)/H2VO4-/(2R,3R) tartrate system and X-ray crystal structures of Na4[V4O8(rac-tart)2].12H2O and (NEt4)4[V4O8((R,R)-tart)2].6H2O (tart = C4H2O6(4-)). AB - A study of the aqueous H3O+(OH-)/H2VO4-/(2R,3R)-tartrate system has been performed at 273 K in a 1.0 mol/L Na+(Cl-) ionic medium using 51V NMR spectroscopy. In this relatively complicated system, more than 12 different species were observed. Ligand concentration, vanadate concentration, and pH variation studies were carried out, particularly for the range of pH 5.8-8.0 and for pH 2.4. Chemical shifts, vanadium-ligand stoichiometry, and also composition and formation constants for some, but not all, species are given. Despite some reduction of vanadium(V) to vanadium(IV) in an acidic medium at pH approximately 2.4, the stoichiometries of the principal species in solution at this pH were determined. Electrospray ionization mass spectra for some solutions were obtained and were in accordance with the conclusions drawn from the speciation studies. A series of crystalline vanadium(V) tartrato complexes M4[V4O8(tart)2].aq were also prepared and characterized. X-ray diffraction studies of Na4[V4O8(rac tart)2].12H2O (1) and (NEt4)4[V4O8((R,R)-tart)2].6H2O (2) revealed unique tetranuclear [V4O8(tart)2]4- ions for which the {V4O4} rings have boat conformations. PMID- 17419618 TI - Nature of Cp*MoO2+ in water and intramolecular proton-transfer mechanism by stopped-flow kinetics and density functional theory calculations. AB - A stopped-flow study of the Cp*MoO3- protonation at low pH (down to zero) in a mixed H2O-MeOH (80:20) solvent at 25 degrees C allows the simultaneous determination of the first acid dissociation constant of the oxo-dihydroxo complex, [Cp*MoO(OH)2]+ (pKa1 = -0.56), and the rate constant of its isomerization to the more stable dioxo-aqua complex, [Cp*MoO2(H2O)]+ (k-2 = 28 s 1). Variable-temperature (5-25 degrees C) and variable-pressure (10-130 MPa) kinetics studies have yielded the activation parameters for the combined protonation/isomerization process (k-2/Ka1) from Cp*MoO2(OH) to [Cp*MoO2(H2O)]+, viz., DeltaH++= 5.1 +/- 0.1 kcal mol-1, DeltaS++ = -37 +/- 1 cal mol-1 K-1, and DeltaV++ = -9.1 +/- 0.2 cm3 mol-1. Computational analysis of the two isomers, as well as the [Cp*MoO2]+ complex resulting from the dissociation of water, reveals a crucial solvent effect on both the isomerization and the water dissociation energetics. Introducing a solvent model by the conductor-like polarizable continuum model and especially by explicitly inclusion of up to three water molecules in the calculations led to the stabilization of the dioxo-aqua species relative to the oxo-dihydroxo isomer and to the substantial decrease of the energy cost for the water dissociation process. The presence of a water dissociation equilibrium is invoked to account for the unusually low effective acidity (pKa1' = 4.19) of the [Cp*MoO2(H2O)]+ ion. In addition, the computational study reveals the positive role of external water molecules as simultaneous proton donors and acceptors, having the effect of dramatically lowering the isomerization energy barrier. PMID- 17419619 TI - Iron(II) complexes with amide-containing macrocycles as non-heme porphyrin analogues. AB - Iron(II) complexes of macrocyclic pentadendate ligands 3,6,9,12,18 pentaazabicyclo[12.3.1]octadeca-1(18),14,16-triene-2,13-dione (H2pydioneN5) and 16-chloro-3,6,9,12,18-pentaazabicyclo[12.3.1]octadeca-1(18),14,16-triene-2,13 dione (H2pyCldioneN5) were synthesized and fully characterized. Complexes with one or two deprotonated amide groups of H2pydione were both isolated. In the former case the metal ion has a distorted octahedral coordination sphere; in the latter case the complex adopts a pentagonal-bipyramidal geometry. NMR experiments show that the protonation state of the ligand is preserved in a dimethyl sulfoxide (DMSO) solution. The complexes maintain a high-spin state even at low temperatures. Detailed kinetic studies of oxygenation of the iron(II) complexes showed that the deprotonation state of the complex has a profound effect on the reactivity with dioxygen. Oxygenation of the dideprotonated complex of iron(II), Fe(pydioneN5), in aprotic solvents proceeds via a path that is analogous to that of iron(II) porphyrins: via iron(III) superoxo and diiron(III) peroxo species, as evidenced by the spectral changes during the reaction, which is second-order in the concentration of the iron(II) complex, and with an inverse dependence of the reaction rate on the concentration of dioxygen. The final products of oxygenation are crystallographically characterized iron(III) mu-oxo dimers. We have also found that the presence of 1-methylimidazole stabilizes the diiron peroxo intermediate. The reaction of Fe(pydioneN5) with dioxygen in methanol is distinctly different under the same conditions. The reaction is first-order in both iron(II) complex and dioxygen, and no intermediate is spectroscopically observed. Similar behavior was observed for the monodeprotonated complex Fe(HpydioneN5)(Cl). The presence of an accessible proton either from the solvent (reactions in methanol) or from the complex itself (in Fe(HpydioneN5)(Cl)) proves sufficient to alter the oxygenation pathway in these macrocyclic systems, which is reminiscent of the properties of iron(II) porphyrin complexes. The new amidopyridine macrocycles can be considered as new members of the "expanded porphyrin analogue" family. The expansion of the cavity provides control over the spin state and availability of protons. These macrocyclic systems also allow for easy synthetic modifications, paving the way to new, versatile metal complexes. PMID- 17419620 TI - An unprecedented polyborate ester anion: X-ray diffraction studies on [1,8 C10H6(NMe2)2H][B5O6(OMe)4]. AB - The slow hydrolysis of B(OMe)3 in a CH2Cl2 solution in the presence of 1,8 C10H6(NMe2)2 (5:1 ratio) led to the formation of the novel isolated pentaborate ester anion [B5O6(OMe)4]-, which was characterized by a single-crystal X-ray diffraction study as the salt [1,8-C10H6(NMe2)2H][B5O6(OMe)4]. PMID- 17419621 TI - [Mo(Tm(Me))(O)2Cl]: an alternative functional model of sulfite oxidase. AB - The hydrotris(methimazolyl)borate anion (TmMe) has been used to synthesize an alternative functional model ([Mo(TmMe)(O)2Cl]) of the metalloenzyme sulfite oxidase. It has been shown that the complex undergoes oxygen atom transfer chemistry and that it performs the primary function of the enzyme, sulfite oxidation. A method using ion chromatography has been developed to definitively prove that sulfite is oxidized to sulfate. Employment of a soft tripodal ligand has allowed us to tune the redox potentials of our complex so that they are significantly closer to those reported for sulfite oxidase. PMID- 17419624 TI - Are carboxyl groups the most acidic sites in amino acids? Gas-phase acidity, H/D exchange experiments, and computations on cysteine and its conjugate base. AB - Hydrogen-deuterium exchange experiments were carried out on the conjugate base of cysteine with four different deuterated alcohols. Three H/D exchanges are observed to take place in each case, and a relay mechanism which requires the SH and CO2H groups to have similar acidities and subsequently proceeds through a zwitterionic intermediate is proposed. Gas-phase acidity measurements also were carried out in a quadrupole ion trap using the extended kinetic method and in a Fourier transform mass spectrometer by an equilibrium determination. The results are in excellent accord with each other and high-level ab initio and density functional theory calculations and indicate that the side-chain thiol in cysteine is more acidic than the carboxyl group by 3.1 kcal mol-1. Deprotonated cysteine is thus predicted to be a thiolate ion. A zwitterionic species also was located on the potential energy surface, but it is energetically unfavorable (+10.1 kcal mol-1). PMID- 17419625 TI - Secretory phospholipase A2 hydrolysis of phospholipid analogues is dependent on water accessibility to the active site. AB - A new and unnatural type of phospholipids with the head group attached to the 2 position of the glycerol backbone has been synthesized and shown to be a good substrate for secretory phospholipase A2 (sPLA2). To investigate the unexpected sPLA2 activity, we have compared three different phospholipids by using fluorescence techniques and HPLC, namely: (R)-1,2-dipalmitoyl-glycero-3 phosphocholine (hereafter referred to as 1R), (R)-1-O-hexadecyl-2-palmitoyl glycero-3-phoshocholine (2R), and (S)-1-O-hexadecyl-3-palmitoyl-glycero-2 phosphocholine (3S). Furthermore, to understand the underlying mechanisms for the observed differences, we have performed molecular dynamics simulations to clarify on a structural level the substrate specificity of sPLA2 toward phospholipid analogues with their head groups in the 2-position of the glycerol backbone. We have studied the lipids above 1R, 2R, and 3S as well as their enantiomers 1S, 2S, and 3R. In the simulations of sPLA2-1S and sPLA2-3R, structural distortion in the binding cleft induced by the phospholipids showed that these are not substrates for sPLA2. In the case of the phospholipids 1R, 2R, and 3S, our simulations revealed that the difference observed experimentally in sPLA2 activity might be caused by reduced access of water molecules to the active site. We have monitored the number of water molecules that enter the active site region for the different sPLA2-phospholipid complexes and found that the probability of a water molecule reaching the correct position such that hydrolysis can occur is reduced for the unnatural lipids. The relative water count follows 1R > 2R > 3S. This is in good agreement with experimental data that indicate the same trend for sPLA2 activity: 1R > 2R > 3S. PMID- 17419626 TI - Foldamers as reactive sieves: reactivity as a probe of conformational flexibility. AB - A series of m-phenyleneethynylene (mPE) oligomers modified with a dimethylaminopyridine (DMAP) unit were treated with methyl sulfonates of varying sizes and shapes, and the relative reactivities were measured by UV spectrophotometry. Using a small-molecule DMAP analogue as a reference, each of the methyl sulfonates was shown to react at nearly identical rate. In great contrast, oligomers that are long enough to fold, and hence capable of binding the methyl sulfonate, experience rate enhancements of 18-1600-fold relative to that of the small-molecule analogue, depending on the type of alkyl chain attached to the guest. Three different oligomer lengths were studied, with the longest oligomers exhibiting the fastest rate and greatest substrate specificity. Even large, bulky guests show slightly enhanced methylation rates compared to that with the reference DMAP, which suggests a dynamic nature to the oligomer's binding cavity. Several mechanistic models to describe this behavior are discussed. PMID- 17419627 TI - 4-Aryl-1,3,2-oxathiazolylium-5-olates as pH-controlled NO-donors: the next generation of S-nitrosothiols. AB - S-Nitrosothiols (RSNOs) are important exogenous and endogenous sources of nitric oxide (NO) in biological systems. A series of 4-aryl-1,3,2-oxathiazolylium-5 olates derivatives with varying aryl para-substituents (-CF3, -H, -Cl, and -OCH3) were synthesized. These compounds were found to release NO under acidic condition (pH = 5). The decomposition pathway of the aryloxathiazolyliumolates proceeded via an acid-catalyzed ring-opening mechanism after which NO was released and an S centered radical was generated. Electron paramagnetic resonance (EPR) spin trapping studies were performed to detect NO and the S-centered radical using the spin traps of iron(II) N-methyl-D-glucamine dithiocarbamate [(MGD)2-FeII] and 5,5 dimethyl-1-pyrroline N-oxide (DMPO). Also, EPR spin trapping and UV-vis spectrophotometry were used to analyze the effect of aryl para substitution on the NO-releasing property of aryloxathiazolyliumolates. The results showed that the presence of an electron-withdrawing substituent such as -CF3 enhanced the NO releasing capability of the aryloxathiazolyliumolates, whereas an electron donating substituent like methoxy (-OCH3) diminished it. Computational studies using density functional theory (DFT) at the PCM/B3LYP/6-31+G**//B3LYP/6-31G* level were used to rationalize the experimental observations. The aryloxathiazolyliumolates diminished susceptibility to reduction by ascorbate or gluthathione, and their capacity to cause vasodilation as compared to other S nitrosothiols suggests potential application in biological systems. PMID- 17419628 TI - Solid-phase synthesis of oxathiocoraline by a key intermolecular disulfide dimer. PMID- 17419629 TI - Macrocyclic beta-sheet peptides that mimic protein quaternary structure through intermolecular beta-sheet interactions. AB - This paper reports the design, synthesis, and characterization of a family of cyclic peptides that mimic protein quaternary structure through beta-sheet interactions. These peptides are 54-membered-ring macrocycles comprising an extended heptapeptide beta-strand, two Hao beta-strand mimics [JACS 2000, 122, 7654] joined by one additional alpha-amino acid, and two delta-linked ornithine beta-turn mimics [JACS 2003, 125, 876]. Peptide 3a, as the representative of these cyclic peptides, contains a heptapeptide sequence (TSFTYTS) adapted from the dimerization interface of protein NuG2 [PDB ID: 1mio]. 1H NMR studies of aqueous solutions of peptide 3a show a partially folded monomer in slow exchange with a strongly folded oligomer. NOE studies clearly show that the peptide self associates through edge-to-edge beta-sheet dimerization. Pulsed-field gradient (PFG) NMR diffusion coefficient measurements and analytical ultracentrifugation (AUC) studies establish that the oligomer is a tetramer. Collectively, these experiments suggest a model in which cyclic peptide 3a oligomerizes to form a dimer of beta-sheet dimers. In this tetrameric beta-sheet sandwich, the macrocyclic peptide 3a is folded to form a beta-sheet, the beta-sheet is dimerized through edge-to-edge interactions, and this dimer is further dimerized through hydrophobic face-to-face interactions involving the Phe and Tyr groups. Further studies of peptides 3b-3n, which are homologues of peptide 3a with 1-6 variations in the heptapeptide sequence, elucidate the importance of the heptapeptide sequence in the folding and oligomerization of this family of cyclic peptides. Studies of peptides 3b-3g show that aromatic residues across from Hao improve folding of the peptide, while studies of peptides 3h-3n indicate that hydrophobic residues at positions R3 and R5 of the heptapeptide sequence are important in oligomerization. PMID- 17419630 TI - Protein folding kinetics: barrier effects in chemical and thermal denaturation experiments. AB - Recent experimental work on fast protein folding brings about an intriguing paradox. Microsecond-folding proteins are supposed to fold near or at the folding speed limit (downhill folding), but yet their folding behavior seems to comply with classical two-state analyses, which imply the crossing of high free energy barriers. However, close inspection of chemical and thermal denaturation kinetic experiments in fast-folding proteins reveals systematic deviations from two-state behavior. Using a simple one-dimensional free energy surface approach we find that such deviations are indeed diagnostic of marginal folding barriers. Furthermore, the quantitative analysis of available fast-kinetic data indicates that many microsecond-folding proteins fold downhill in native conditions. All of these proteins are then promising candidates for an atom-by-atom analysis of protein folding using nuclear magnetic resonance.1 We also find that the diffusion coefficient for protein folding is strongly temperature dependent, corresponding to an activation energy of approximately 1 kJ.mol-1 per protein residue. As a consequence, the folding speed limit at room temperature is about an order of magnitude slower than the approximately 1 micros estimates from high temperature T-jump experiments. Our analysis is quantitatively consistent with the available thermodynamic and kinetic data on slow two-state folding proteins and provides a straightforward explanation for the apparent fast-folding paradox. PMID- 17419631 TI - Polymerase-directed synthesis of 2'-deoxy-2'-fluoro-beta-D-arabinonucleic acids. PMID- 17419632 TI - Sapphyrin-nanotube assemblies. AB - Single wall carbon nanotubes (SWNTs) bind strongly to sapphyrins, quintessential pentapyrrolic "expanded porphyrin" macrocycles, through donor-acceptor stacking interactions. The specific use of a functionalized sapphyrin diol yields stable water-suspendable nanotubes and also permits the formation of well-defined assemblies in ionic liquids. The absorption and steady-state fluorescence spectra of the resulting noncovalently functionalized nanotube complexes have been analyzed in aqueous media and ionic liquids, yielding a description of the photophysical properties of the nanotube-sapphyrin complexes as donor-acceptor species for light-harvesting. PMID- 17419635 TI - Induction of autophagy and apoptosis by the extract of Solanum nigrum Linn in HepG2 cells. AB - Solanum nigrum L. (SN) has been used in traditional folk medicine to treat different cancers. It is also used as a hepatoprotective and anti-inflammatory agent. In this study, we demonstrated that the extract of SN (SNE) induced a strong cytotoxic effect toward HepG2 cells but much less to Chang liver and WRL 68 cells. The mechanisms of the cytotoxic effect were concentration-dependent. High doses of SNE (2 and 5 mg/mL) induced apoptotic cell death in HepG2 cells, as evidenced by increases in the expressions of p-JNK and Bax, mitochodrial release of cytochrome c, and caspase activation. On the other hand, cells treated with low concentrations of SNE (50-1000 microg/mL) revealed morphological and ultrastructural changes of autophagocytic death under electron microscopic observation. Furthermore, these cells showed increased levels of autophagic vacuoles and LC3-I and LC3-II proteins, specific markers of autophagy. The levels of Bcl-2 and Akt that have been implicated in the down-regulation of autophagy were decreased upon SNE treatment. Taken together, these findings indicate that SNE induced cell death in hepatoma cells via two distinct antineoplastic activities of SNE, the ability to induce apoptosis and autophagocytosis, therefore suggesting that it may provide leverage to treat liver cancer. PMID- 17419636 TI - Ethyl hexanoate transfer modeling in carrageenan matrices for determination of diffusion and partition properties. AB - Aroma compound properties in food matrices, such as volatility and diffusivity, have to be determined to understand the effect of composition and structure on aroma release and perception. This work illustrates the use of mass transfer modeling to identify diffusion and partition properties of ethyl hexanoate in water and in carrageenan matrices with various degrees of structure. The comparison of results obtained with a diffusive model to those obtained with a convective model highlights the importance of considering the appropriate transfer mechanism. Modeling of the preliminary experimental steps ensures correct estimation of the conditions for the main aroma release step. The obtained values of partition and diffusion coefficients are in agreement with those found in the literature (either experimentally determined or predicted by theoretical equations) and demonstrate that the structure level of carrageenan matrices has little influence on diffusion properties of ethyl hexanoate (less than 20%). PMID- 17419637 TI - Polymerase Chain Reaction assay for the detection of Kudoa paniformis and Kudoa thyrsites in Pacific Hake (Merluccius productus). AB - Myoliquefaction of Pacific hake has been attributed to proteolytic action associated with parasitic infection. Among the two infecting species of Kudoa, Kudoa paniformis and Kudoa thyrsites, the former is reported to be more virulent for the "soft flesh" phenomenon in Pacific hake. The objective of this research was to develop a sensitive and specific polymerase chain reaction (PCR) assay to detect infection of hake by K. paniformis. Primers based on specific regions ( approximately 1562 bp) of the small subunit ribosomal DNA of K. paniformis successfully amplified the target DNA segments from both spore and muscle extracted DNA templates. DNA sequencing confirmed the veracity of this method to distinguish parasitic infection by K. paniformis versus K. thyrsites. The established PCR method was applied to investigate Kudoa infection in 44 Pacific hake samples using DNA extracted from muscle and/or spores, and the results were compared to infection evaluated by microscopic examination of extracted spores. PMID- 17419638 TI - Efficiency of natural phenolic compounds regenerating alpha-tocopherol from alpha tocopheroxyl radical. AB - Benzoic acid-derived phenolics (p-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, vanillic acid, syringic acid, and gallic acid) and the polyphenols epicatechin and epigallocatechin gallate (EGCG) were evaluated for their efficiency in regenerating alpha-tocopherol from alpha-tocopheroxyl radical in comparison with ascorbyl palmitate, which is known to regenerate alpha-tocopherol. Ethanolic solutions of phenolic compounds were added to a homogeneous hexane medium containing alpha-tocopheroxyl radical generated by reaction of alpha-tocopherol in molar excess with 1,1-diphenyl-2-picrylhydrazyl radical, and the alpha tocopheroxyl radical was monitored by electron spin resonance spectroscopy. p Hydroxybenzoic acid, vanillic acid, and syringic acid (400 microM) did not exhibit a significant effect on alpha-tocopheroxyl radical concentration (0.6-0.7 microM). In contrast, 3,4-dihydroxybenzoic acid and gallic acid were able to reduce the concentration of alpha-tocopheroxyl radical by 16 and 64%, respectively. Epicatechin showed a reduction of alpha-tocopheroxyl radical similar to gallic acid, and EGCG and ascorbyl palmitate were the most effective, reducing alpha-tocopheroxyl radical completely at a much lower phenolic concentration (66.7 microM). The moles of alpha-tocopheroxyl radical reduced per mole of ascorbyl palmitate (0.93), EGCG (0.066), gallic acid (4.3 x 10(-4)), and epicatechin (4.5 x 10(-4)) were determined, and the logarithm of these stoichoimetric ratios showed a negative linear relationship with the bond dissociation enthalpies of the O-H bond of the phenolics. The relative capacity to reduce alpha-tocopheroxyl radical was found to be ascorbyl palmitate (2142) > EGCG (151) > gallic acid (1) approximately epicatechin (1). PMID- 17419639 TI - Influence of emulsifiable concentrate formulation on the physical properties of the fluid, spray characteristics, and insecticide deposits on stored grains. AB - In stored grains, smaller depositions and great variations with regard to theoretical insecticide doses are frequently found. The objective of this work was to evaluate the influence of the emulsifiable concentrate formulation on the physical properties of the liquid, volumetric distribution, droplet spectrum, and insecticide deposits on stored grains. To determine its physical properties, the applied mix was prepared at a concentration of 0.4% of commercial product. Volumetric distribution was used as an evaluation parameter in a model TJ-60 8002EVS hydraulic nozzle study, and clean water and insecticidal mix were used as test fluids. After the effective swath width (esw) had been determined for both fluids, an application system was built to apply theoretical concentrations of 10 and 0.5 mg kg-1 of fenitrothion and esfenvalerate, respectively. Mix viscosity was 82% higher than water viscosity; conversely, surface tension in the mix corresponded to 49% of the water surface tension value. For water and mix, esw values were 0.425 and 0.60 m, respectively. Deposits of both insecticides at the 0.60 m esw were significantly higher (P<0.05) than deposits at the 0.425 m esw. The results obtained demonstrate the great influence of emulsifiable concentrate formulation on the physical properties of the fluid, spray characteristics, and insecticide deposits on stored grains. PMID- 17419640 TI - Detection of hexabromocyclododecane and its metabolite pentabromocyclododecene in chicken egg and fish from the official food control. AB - During routine gas chromatography with electron capture detection (GC/ECD) analysis of chicken eggs, we observed that the most prominent peak in some samples did not match the retention time of any of the food contaminants screened. Subsequent GC coupled with mass spectrometry (GC/MS) studies clarified that the mass spectrum of the peak was very similar to hexabromocyclododecane (HBCD), which was also identified by GC/MS in the egg. The unknown compound was positively identified as pentabromocyclododecene (PBCDE), a metabolite of HBCD detected for the first time in foodstuffs. Studies of the analytical method used for the analysis of pesticides and contaminants showed that this cleanup method was suitable for the determination of HBCD and PBCDE, but storage of sample extracts resulted in the loss of HBCD when the sample extracts were not sufficiently purified. The concentrations of HBCD and PBCDE in the high polluted sample were 2.0 and 3.6 mg/kg egg fat. HBCD and PBCDE were also detected in two additional eggs at lower levels (<0.15 mg/kg), whereas 75 eggs did not contain these compounds (<0.02 mg/kg). We also detected HBCD and PBCDE in two samples of whitefish (Coregonus sp.), while an eel sample (Anguilla anguilla) positively tested for HBCD did not contain PBCDE. Surprisingly, the potential metabolite of HBCD, PBCDE, has not been detected before in any food or environmental sample. The present results indicate that more attention should be paid to the detection of HBCD and its metabolite PBCDE in chicken eggs. PMID- 17419641 TI - Chemical and physical stability of citral and limonene in sodium dodecyl sulfate chitosan and gum arabic-stabilized oil-in-water emulsions. AB - Citral and limonene are the major flavor components of citrus oils. Both of these compounds can undergo chemical degradation leading to loss of flavor and the formation of undesirable off-flavors. Engineering the interface of emulsion droplets with emulsifiers that inhibit chemical reactions could provide a novel technique to stabilize citral and limonene. At present, emulsified flavor oils are usually stabilized by gum arabic (GA), which is a naturally occurring polysaccharide-protein complex. The objective of this study was to examine if citral and limonene were more stable in emulsions stabilized with a sodium dodecyl sulfate (SDS)-chitosan complex than GA. Citral degraded less in GA stabilized than in SDS-chitosan-stabilized emulsions at pH 3.0. However, SDS chitosan-stabilized emulsions were more effective at retarding the formation of the citral oxidation product, p-cymene, than GA-stabilized emulsions. Limonene degradation and the formation of limonene oxidation products, limonene oxide and carvone, were lower in the SDS-chitosan- than GA-stabilized emulsions at pH 3.0. The ability of an SDS-chitosan multilayer emulsifier system to inhibit the oxidative deterioration of citral and limonene could be due to the formation of a cationic and thick emulsion droplet interface that could repel prooxidative metals, thus decreasing prooxidant-lipid interactions. PMID- 17419642 TI - Determination of folate concentrations in diverse potato germplasm using a trienzyme extraction and a microbiological assay. AB - Folate deficiency is a leading cause of birth defects and is implicated in several other diseases. We are interested in how much folate concentrations vary among potato germplasm. We determined total folate concentrations of potato tubers from 67 cultivars, advanced breeding lines, or wild species. Folates were extracted by a tri-enzyme treatment and analyzed by using a Lactobacillus rhamnosus microbiological assay. Folate concentrations varied from 521 +/- 96 to 1373 +/- 230 ng/g dry weight and were genotype and location dependent. The highest folate concentrations were mostly found in color-fleshed potatoes. Variations of folate concentrations within either color- or white-fleshed tubers were similar ( approximately 2-fold). Skin contained approximately 30% higher folate concentrations than flesh. Storage of tubers for 7 months generally led to an increase in folate contents. Semiquantitative RT-PCR analyses showed that higher folate contents were correlated with lower mRNA expression of some folate genes. PMID- 17419643 TI - Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples. AB - Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health. PMID- 17419644 TI - Oxidation products of alpha-farnesene associated with superficial scald development in d'Anjou pear fruits are conjugated trienols. AB - Conjugated triene (CT) oxidation products of the acyclic sesquiterpene (E,E) alpha-farnesene are thought to induce development of the physiological storage disorder superficial scald in apple and pear fruits of susceptible cultivars. CTs that accumulate in peel tissues of Granny Smith and Delicious apples after several weeks of cold storage are known to be conjugated trienols (CTols) rather than the corresponding hydroperoxides produced by in vitro autoxidation of alpha farnesene. Here, it is shown that CTols are also the predominant in vivo oxidation products of alpha-farnesene in cold-stored pear fruit of the highly scald-susceptible d'Anjou cultivar. Analysis by octadecylsilane reversed phase high-performance liquid chromatography with diode array detection, gas chromatography with flame ionization detection, gas chromatography-mass spectrometry, and proton nuclear magnetic resonance spectroscopy identified 3,7,11-trimethyldodeca-1,3(E),5(E),10-tetraen-7-ol as the major CT in hexane extracts of peel tissue from d'Anjou pears stored for 3-5 months in air at -1 degrees C. The possible origins of CTols in apples and pears and the hypothesized role of these oxidation products of alpha-farnesene in the induction of scald are discussed. . PMID- 17419645 TI - Allium discoloration: the color-forming potential of individual thiosulfinates and amino acids: structural requirements for the color-developing precursors. AB - Structural requirements for compounds involved in Allium discoloration have been investigated in detail. The abilities of all 20 protein amino acids and six naturally occurring 1-propenyl-containing thiosulfinates to form the pigments have been studied. Furthermore, several analogues of these thiosulfinates were prepared by synthesis, and their color-forming abilities were evaluated, together with those of various amino compounds. It has been found that an unsubstituted primary amino group and a free carboxyl group are essential structural features required for amino compounds to be able to generate the pigments. Out of the thiosulfinate analogues tested, only those containing at least a three-carbon chain with the beta-carbon bearing a hydrogen atom yielded the pigments after reacting with glycine. Thiosulfonates, sulfoxides, sulfides, and disulfides did not form any colored products when mixed with glycine. The pH optimum for pigment formation has been found to be between 5.0 and 6.0 for all thiosulfinates tested. PMID- 17419646 TI - Determination of aloesin and aloeresin A for the detection of aloe in beverages. AB - This work describes a sensitive high-performance liquid chromatography (HPLC) method for the quantification of aloesin and aloeresin A in alcoholic beverages containing aloe as a flavoring agent. The compounds were prepared from Aloe ferox juice. Sephadex LH20 and ion-exchange resin AG1X2 column chromatography were used for aloesin. Aloeresin A was obtained by Sephadex LH20 and silica gel column chromatography followed by purification on Discovery DSC-18 solid-phase extraction tubes. A 98 mg amount of aloesin (>99% purity) and 34 mg of aloeresin A (>98% purity) were recovered from 2.5 g of aloe juice. The HPLC method was validated, and intra- and interday performances were established. In-house validation was carried out by analyzing samples of beverages with and without aloe as a flavoring agent. PMID- 17419647 TI - Electrosynthesis of symmetric and highly conjugated benzofuran via a unique ECECCC electrochemical mechanism: evidence for predominance of electrochemical oxidation versus intramolecular cyclization. AB - Electrochemical oxidation of hydroquinone, catechol, and some of their monosubstituted derivatives has been studied in the presence of 3-hydroxy-1H phenalen-1-one (2) as a nucleophile in water/acetonitrile (80/20) solutions using cyclic voltammetry and controlled-potential coulometry methods. The results revealed that quinones derived from oxidation of hydroquinones and catechols participate in Michael addition reactions with 2. The formed adducts convert to the corresponding benzofuran derivatives via different mechanisms. In this work, we derived a variety of products with good yields using controlled potential electrochemical oxidation at a graphite electrode in an undivided cell. PMID- 17419648 TI - A facile approach to the synthesis of chiral 2-substituted benzofurans. AB - An effective route to chiral optically active 2-substituted benzofurans directly from carboxylic acids is reported. This procedure, which allows the preparation of alpha-alkyl-2-benzofuranmethanamines from N-protected alpha-amino acids without sensible racemization phenomena, proceeds in good yields under mild conditions with the help of microwave irradiation. PMID- 17419649 TI - Enantio- and regioselective heck-type reaction of arylboronic acids with 2,3 dihydrofuran. AB - Reported herein is a protocol for the enantioselective Pd(II)-catalyzed Heck-type reaction between arylboronic acids and 2,3-dihydrofuran. The highest chemical and optical yields were obtained when a Pd(OAc)2/(R)-MeO(biphenylphosphine) or a Pd(OAc)2/(R)-(2,2'-bis(diphenylphosphino)-1,1'-binaphthyl) catalyst and a Cu(OAc)2 reoxidant were employed. PMID- 17419650 TI - First example of direct RuO4-catalyzed oxidation of isoxazolidines to 3 isoxazolidones. AB - RuO2/NaIO4 oxidation of 3-unsubstituted isoxazolidines, under ethyl acetate/water biphasic conditions, affords 3-isoxazolidones in good yields. The methodology can be used on both racemic and optically active isoxazolidines. PMID- 17419651 TI - Copper-mediated synthesis of tertiary diaryl squaramides. AB - Tertiary aryl squaramides were synthesized by using copper catalyzed C-N bond formation with L-proline as the ligand. Symmetrical diaryl squaramides could be prepared in a one-pot reaction by using excess aryl bromide with varying yields. Unsymmetrical derivatives were prepared by sequential arylation. Yields of the diarylated products were highly sensitive to steric effects. PMID- 17419652 TI - Novel method for measuring the adhesion energy of vesicles. AB - Adhering vesicles with osmotically stabilized volume are studied with Monte Carlo simulations and optical microscopy. The simulations are used to determine the dependence of the adhesion area on the vesicle volume, the surface area, the bending rigidity, the adhesion energy per membrane area, and the adhesion potential range. The simulation results lead to a simple functional expression that is supplemented by a correction term for gravity effects. The obtained equation provides a new tool to analyze optical microscopy data and, thus, to measure the adhesion energy per area by analyzing the geometry of the adhering vesicle. The method can be applied in the weak and ultra-weak adhesion regime, where the adhesion energy per area is below 10(-6) J/m(2). By comparing the shapes of adhering vesicles with different reduced volumes, the bending rigidity can be estimated as well. The new approach is applied to experimental data for lipid vesicles on (i) an untreated and (ii) a monolayer-coated glass surface, providing ultra-weak and weak adhesion strength, respectively. PMID- 17419653 TI - Specific adsorption of cytochrome C on cardiolipin-glycerophospholipid monolayers and bilayers. AB - In this study, we examined the adsorption of cytochrome c (cyt c) on monolayers and liposomes formed from (i) pure 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), or cardiolipin (CL) and on (ii) the more thermodynamically stable binary mixtures of POPE/CL (0.8:0.2 mol/mol) and POPC/CL (0.6:0.4 mol/mol). Constant surface pressure experiments showed that the maximum and minimum interactions occurred in the pure CL (anionic phospholipid) and the pure POPE (zwitterion) monolayers, respectively. Observation by atomic force microscopy (AFM) of the images of Langmuir-Blodgett (LB) films extracted at 30 mN m-1 suggests that the different interactions of cyt c with POPE/CL and the POPC/CL monolayers could be due to lateral phase separation occurring in the POPE/CL mixture. The competition between 8-anilino-1-naphthalene sulfonate (ANS) and cyt c for the same binding sites in liposomes that have identical nominal compositions with respect to those of the monolayers was used to obtain binding parameters. In agreement with the monolayer experiments, the most binding was observed in POPE/CL liposomes. All of our observations strongly support the existence of selective adsorption of cyt c on CL, which is modulated differently by different neutral phospholipids (POPE and POPC). PMID- 17419654 TI - Competitive adsorption of nonionic surfactant and nonionic polymer on silica. AB - Competitive adsorption of the nonionic polymer poly(ethylene oxide) (PEO) and the nonionic surfactant of the type poly(ethylene oxide) alkyl ether from aqueous solutions on a silica surface is examined. From one-component solutions, both species readily adsorb onto silica and, in the bulk of mixed (two-component) solutions, polymer-surfactant complexes are not observed. Because both species bind by the same mechanism to silica, subtle differences in layer structure, or other species-specific parameters, determine whether one or both of the species will adsorb. It was found that various surfactants can displace PEO up to a certain critical molecular weight. Surfactants with a high aggregation number, in bulk and on the surface, can displace PEO with a higher molar mass than surfactants with a low aggregation number. As the molar mass of the polymer increases, the time a surfactant needs to completely displace the polymer increases. We can explain both the existence of the critical molar mass and the decrease in adsorption kinetics with a shift in the critical surface association concentration (CSAC). PMID- 17419655 TI - Kinetics of channel formation in bilayer lipid membranes (BLMs) and tethered BLMs: monazomycin and melittin. AB - The kinetics of channel formation by the polyene-like antibiotic monazomycin, both in a bilayer lipid membrane (BLM) and in a tethered BLM (tBLM), and by the peptide melittin in a tBLM, is investigated. Stepping the applied potential from a value at which channels are not formed to one at which they are formed yields current vs time curves that are sigmoidal on a BLM, while they show a maximum on a tBLM; in the latter case, sigmoidal curves are obtained by plotting the charge against time. These curves are interpreted on the basis of a general kinetic model, which accounts for the potential-dependent penetration of adsorbed monomeric molecules into the lipid bilayer, followed by their aggregation with channel formation by a mechanism of nucleation and growth. In the case of monazomycin, which is present in the solution in the form of relatively hydrophilic clusters and is adsorbed as such on top of the lipid bilayer, penetration into the bilayer following a potential jump is assumed to be preceded by a potential-independent disaggregation of the adsorbed clusters into adsorbed monomers. PMID- 17419656 TI - Optimizing biosensing properties on undecylenic Acid-functionalized diamond. AB - The optimization of biosensing efficiency on a diamond platform depends on the successful coupling of biomolecules on the surface, and also on effective signal transduction in the biorecognition events. In terms of biofunctionalization of diamond surfaces, surface electrochemical studies of diamond modified with undecylenic acid (UA), with and without headgroup protection, were performed. The direct photochemical coupling method employing UA was found to impart a higher density of carboxylic acid groups on the diamond surface compared to that using trifluoroethyl undecenoate (TFEU) as the protecting group during the coupling process. Non-faradic impedimetric DNA sensing revealed that lightly doped diamond gives better signal transduction sensitivity compared to highly doped diamond. PMID- 17419657 TI - Preparation of poly(ethylene glycol)-modified poly(amido amine) dendrimers encapsulating gold nanoparticles and their heat-generating ability. AB - Loading of HAuCl4 in poly(amido amine) G4 dendrimers having poly(ethylene glycol) (PEG) grafts at all chain ends and subsequent reduction with NaBH4 yielded PEG modified dendrimers encapsulating gold nanoparticles (Au NPs) of ca. 2 nm diameter. The Au NPs held in the dendrimers were stable in aqueous solutions and dissolved readily, even after freeze-drying. Despite their small particle size, the heat-generating ability of Au NPs held in the dendrimer was comparable to that of widely used Au NPs with ca. 11 nm diameter under visible light irradiation. The observed excellent colloidal stability, high heat-generating ability and their biocompatible surface confirm that the PEG-modified dendrimers encapsulating Au NPs are a promising tool for photothermal therapy and imaging. PMID- 17419658 TI - Optical absorption of DNA-carbon nanotube structures. AB - We measured the UV optical absorption of single-stranded DNA bound to single walled carbon nanotubes (DNA/SWNT). The nucleotide absorbance from DNA/SWNT provides the first experimental confirmation that DNA binds to nanotubes through pi-stacking. Because the hypochromic absorbance typical of pi-stacked structures are expected to occur primarily for DNA dipole transitions that lie along the axis of the optically anisotropic SWNTs, the absorbance changes following binding of DNA to nanotubes reveal the preferred orientation assumed by each of the four bound nucleotides with respect to the nanotube's long axis. PMID- 17419659 TI - 3-D X-ray diffraction imaging with nanoscale resolution using incoherent radiation. AB - A novel approach to X-ray diffraction data analysis for nondestructive determination of the shape of nanoscale particles and clusters in three dimensions is illustrated with representative examples of composite nanostructures. The technique is insensitive to the X-ray coherence, which allows 3-D reconstruction of a modal image without tomographic synthesis and in situ analysis of large (over a several cubic millimeters) volume of material with a spatial resolution of few nanometers, rendering the approach suitable for laboratory facilities. PMID- 17419660 TI - Magnetically actuated nanorod arrays as biomimetic cilia. AB - We present a procedure for producing high-aspect-ratio cantilevered micro- and nanorod arrays of a PDMS-ferrofluid composite material. The rods have been produced with diameters ranging from 200 nm to 1 mum and aspect ratios as high as 125. We demonstrate actuation of these superparamagnetic rod arrays with an externally applied magnetic field from a permanent magnet and compare this actuation with a theoretical energy-minimization model. The structures produced by these methods may be useful in microfluidics, photonic, and sensing applications. PMID- 17419661 TI - The use of music therapy within the SCERTS model for children with Autism Spectrum Disorder. AB - The SCERTS model is a new, comprehensive curriculum designed to assess and identify treatment goals and objectives within a multidisciplinary team of clinicians and educators for children with Autism Spectrum Disorders (ASD). This model is an ongoing assessment tool with resulting goals and objectives derived there from. Because music therapy offers a unique interaction setting for children with ASD to elicit communication skills, music therapists will need to be an integral part of the multidisciplinary assessment team using the SCERTS model which is projected to become the primary nation wide curriculum for children with ASD. The purpose of this paper is to assist music therapists in transitioning to this model by providing an overview and explanation of the SCERTS model and by identifying how music therapists are currently providing clinical services incorporated in the SCERTS Model for children with ASD. In order to formulate comprehensive transitional suggestions, a national survey of music therapists working with clients at risk or diagnosed with ASD was conducted to: (a) identify the areas of SCERTS assessment model that music therapists are currently addressing within their written goals for clients with ASD, (b) identify current music therapy activities that address various SCERTS goals and objectives, and (c) provide demographic information about settings, length, and tools used in music therapy interventions for clients with ASD. PMID- 17419662 TI - The use of music to enhance reading skills of second grade students and students with reading disabilities. AB - The purpose of the present study was to determine the efficacy of using music as a remedial strategy to enhance the reading skills of second-grade students and students who have been identified as having a specific learning disability (SLD) in reading. First, an intensive short-term music curriculum was designed to target reading comprehension and vocabulary skills at the second grade level. The curriculum was then implemented in classrooms at two public schools in the Southeast. Reading skills were evaluated pre and post curriculum intervention via the vocabulary and reading comprehension subtests of the Gates-MacGinitie Reading Test for second grade. Analysis of pre/posttest data revealed that students with a specific disability in reading improved significantly from pre to post on all three subtests: word decoding (p = .04), word knowledge (p = .01), reading comprehension (p = .01), and test total (p = .01). Paired t-tests revealed that for 2nd grade students, both treatment and control classes improved significantly from pre to post on the subtests word decoding, word knowledge, and test total. While both classes made gains from pre to post on the subtest, reading comprehension, neither improved significantly. Analysis of Covariance revealed that the treatment class made greater gains pre to post than the control class on all 3 subtests (Including reading comprehension), and significantly greater gains on the subtest, word knowledge (p = .01). PMID- 17419663 TI - Talking with music teachers about inclusion: perceptions, opinions and experiences. AB - Most information concerning teachers' attitudes regarding inclusion is dated. The present study used the interview methodology to examine issues prevalent in previous studies (e.g., support services) and issues not yet studied (e.g., parent contact, effects on teachers). Research questions focused on (a) information, support, resources, and placements; (b) parent contact and involvement; (c) outcomes on students with disabilities, typical students, and teachers; and (d) teachers' advice. Individual interviews were conducted with 43 teachers (16 elementary, 15 orchestra, & 12 band). Perceived emotional content for teachers' responses was also assessed. Results show that teachers have generally positive attitudes concerning inclusion and their access to support. Attitudes were also positive regarding outcomes for both students with and without disabilities. Several differences and consistencies among the groups lead to questions that merit the study of possible relationships among variables (parent contact, type of support, teacher attitudes). PMID- 17419664 TI - The effect of vision and hearing loss on listeners' perception of referential meaning in music. AB - The purpose of the present study was to examine the effect of vision and hearing loss on listeners' perception of referential meaning in music. Participants were students at a state school for the deaf and blind, and students with typical hearing and vision who attended neighboring public schools (N = 96). The music stimuli consisted of six 37-second randomly ordered excerpts from Saint Saens, Carnival of the Animals. The excerpts were chosen because of their use in similar studies and the composer's clearly intended meaning conveyed in the titles of the excerpts. After allowing for appropriate procedural accommodations for participants with hearing or vision loss, all participants were asked to select the image portrayed by the music. A univariate ANOVA was computed to address the research question, "Do students with vision or hearing loss assign the same visual images to music as students without such sensory losses?" Data were analyzed to examine the effects of sensory condition as well as age and gender. A significant main effect was found for sensory condition, with follow up tests indicating that participants with typical hearing and vision agreed with the composer's intended meaning significantly more often than did participants with vision or hearing loss. No significant main effects were found for gender or age, and no significant interactions were found. Summary data indicated that selected images were more easily identified, or were more difficult to identify across conditions. The data also revealed an order of difficulty and patterns of confusion that were similar across sensory conditions and ages, indicating participant responses were not random, and that some referential meaning in music is conventional. PMID- 17419665 TI - Preservice music teachers' predictions, perceptions, and assessment of students with special needs: the need for training in student assessment. AB - The purpose of the current study was to examine preservice teachers' predictions and perceptions of students with special needs' levels of mastery of specific music education concepts and actual grades achieved by these students using alternative assessments and testing accommodations within two subpopulations: students with emotional and/or behavior disorders (EDBD) and students with acute cognitive delays (ACD). The preservice teachers predicted students within the EDBD class would achieve a significantly higher level of mastery of the music concepts than students within the ACD classroom. After the field experience, however, the preservice teachers' perceptions of all students' levels of mastery increased from prediction scores overall. Additionally, preservice teachers were able to execute testing accommodations and implement successful alternative assessments which gave empirical data on the students' levels of mastery of the music education concepts within the curriculum. Implications for music therapists, as consultants in special education, are discussed. PMID- 17419666 TI - Thioredoxin1 as a negative regulator of cardiac hypertrophy. AB - Excessive reactive oxygen species (ROS) play an important role in the development of cardiac hypertrophy. In contrast, antioxidants scavenge ROS, thereby maintaining the reduced environment of cells and inhibiting hypertrophy in the heart. Thioredoxin1 (Trx1) not only functions as a major antioxidant in the heart but also interacts with important signaling molecules and transcription factors, thereby attenuating cardiac hypertrophy. This review will discuss the molecular mechanisms by which Trx1 exerts antihypertrophic effects in the heart. PMID- 17419667 TI - What is a writing group? Dilemmas of the leader. AB - A writing group can serve to reinforce literary and therapeutic goals. The model outlined here assumes a leader with literary and clinical training, including expertise in group dynamics. The group format is adapted to support exploration of the writer's main problem, the absence of the reader at the moment of writing. The group modifies the writer's "mythical" reader through member interactions with writer and writing. Giving and receiving feedback are central to the group process. The leader's dilemma in a bifocal form like this is to know when and how far to interpret group members' psychological issues. The best rule is to interpret "toward" the group (i.e., to bring up material that can be safely and readily processed there), but to be cautious about interpretation of patterns of early character formation. PMID- 17419668 TI - Anniversary reactions in the therapy group. AB - From Freud onward, the psychoanalytic literature has offered many references to the idea and significance of past time-specific neuroses and to the subterranean lingering of so-called anniversary emotions. Group therapy is particularly effective in surfacing hidden anniversary reactions that often block the course of treatment. Strikingly, groups can perceive buried associations to long past calendar dates such as traumatic birth and death days, holidays, and past seasonal emotionally-laden responses that patients do not consciously recall or dare articulate. This heightened sensitivity to obscured commemorative syndromes can result from the group interactive process. As such the therapy group can aid in surfacing hidden ghosts far more rapidly than individual therapy might achieve. In this paper illustrations of these anniversary reactions arising in groups are presented and the facilitating effects of group process in identifying and resolving these specific resistances are described. PMID- 17419669 TI - A self psychological and relational approach to group therapy for university students with bulimia. AB - Bulimia is a complex disorder that can significantly interfere with psychosocial functioning. It is a particularly significant problem among adolescents and young women in college and university settings. The preponderance of literature on group therapeutic approaches for this disorder is from a cognitive-behavioral perspective. In this paper, we present a psychodynamic group therapy approach for female university students, informed specifically by self psychological and relational frameworks. Through clinical illustrations, we demonstrate how the group provides selfobject experiences, including mirroring, idealizing, twinship, and adversarial experiences, and how the group becomes the arena through which group members can experience, express, and understand emotions. The interpersonal context of the group uniquely provides opportunities for members to examine their bulimic symptoms in relation to interactions that take place outside of or within the group. PMID- 17419670 TI - Psycho-educational group treatment for the severely and persistently mentally ill: how much leader training is necessary? AB - Psycho-educational groups (PEGs) have been shown to be a particularly effective form of treatment for patients classified under the rubric of severely and persistently mentally ill (SPMI). However, recent surveys suggest that certain professionals, such as nurses and clinical psychologists, called on to conduct these groups may have limited to no training. This study tested three methods (self-instructional, workshop, and workshop plus clinical supervision) designed to provide on-the-job training to practicing clinicians. Specifically, training was based on two manuals; one that operationalized the basic knowledge and skills needed to run an effective PEG and the second being a commonly used PEG program for symptom management targeted at SPMI patients. Eight nurses from four adult units at a state psychiatric hospital initially received self-instructional or workshop training and then conducted a 12-session symptom management PEG composed of four to five SPMI patients (e.g., schizophrenia, schizoaffective or major depression disorders). In a second wave of additional training, nurses received workshop training or clinical supervision and then conducted a second PEG. Differences between training methods were assessed by nurse- and patient completed measures that tapped the knowledge and skills emphasized in the symptom management and PEG (nurses only) manuals. Results indicated limited support for the superiority of the workshop method on the nurse measures alone. Implications for training working professionals in group treatments and assessing outcomes with chronic SPMI patients are discussed. PMID- 17419671 TI - Reflections on the writing group. PMID- 17419672 TI - The writing group: establishing protected space. PMID- 17419673 TI - Attachment, group-related processes, and psychotherapy. PMID- 17419674 TI - Where's the salmon? PMID- 17419678 TI - Epimedium-derived phytoestrogen flavonoids exert beneficial effect on preventing bone loss in late postmenopausal women: a 24-month randomized, double-blind and placebo-controlled trial. AB - Epimedium brevicornum maxim, a nonleguminous medicinal plant, has been found to be rich in phytoestrogen flavonoids. Results from a 24-month randomized double blind placebo-controlled clinical trial showed that Epimedium-derived phytoestrogen flavonoids were able to exert beneficial effects on preventing bone loss in late postmenopausal women, without resulting in a detectable hyperplasia effect on the endometrium. INTRODUCTION: We performed a 24-mo randomized double blind placebo-controlled clinical trial for evaluating the effect of the Epimedium-derived phytoestrogen flavonoids (EPFs) on BMD, bone turnover biochemical markers, serum estradiol, and endometrial thickness in postmenopausal women. MATERIALS AND METHODS: One hundred healthy late postmenopausal women, with a natural menopausal history within 10 approximately 18 yr and with a BMD T-score at the lumbar spine between -2 and -2.5 SD, were randomized into EPF treatment group (n = 50; a daily dose of 60 mg Icariin, 15 mg Daidzein, and 3 mg Genistein) or placebo control group (n = 50). All participants received 300 mg element calcium daily. BMD, bone turnover biochemical markers, serum estradiol, and endometrial thickness were measured at baseline and 12 and 24 mo after intervention. RESULTS: Eighty-five participants completed the trial. The patterns of BMD changes were significantly different between the EPF treatment group and placebo control group by repeated-measures ANOVA (p = 0.045 for interaction between time and group at femoral neck; p = 0.006 for interaction between time and group at lumbar spine). BMD was found with a decreased tendency in the placebo control group at 12 (femoral neck: -1.4%, p = 0.104; lumbar spine: -1.7%, p = 0.019) and 24 mo (femoral neck: -1.8%, p = 0.048; lumbar spine: -2.4%, p = 0.002), whereas EPF treatment maintained BMD at 12 (femoral neck: 1.1%, p = 0.285; lumbar spine:1.0%, p = 0.158) and 24 mo (femoral neck: 1.6%, p = 0.148; lumbar spine: 1.3%, p = 0.091). The difference in lumbar spine between the two groups was significant at both 12 (p = 0.044) and 24 mo (p = 0.006), whereas the difference in the femoral neck was marginal at 12 mo (p = 0.061) and significant at 24 mo (p = 0.008). Levels of bone biochemical markers did not change in the placebo control group. In contrast, EPF intervention significantly decreased levels of deoxypyrdinoline at 12 (-43%, p = 0.000) and 24 mo (-39%, p = 0.000), except for osteocalcin at 12 (5.6%, p = 0.530) and 24 mo (10.7%, p = 0.267). A significant difference in deoxypyrdinoline between the two groups was found at both 12 (p = 0.000) and 24 mo (p = 0.001). Furthermore, neither serum estradiol nor endometrial thickness was found to be changed in either groups during the clinical trial. CONCLUSIONS: EPFs exert a beneficial effect on preventing bone loss in late postmenopausal women without resulting in a detectable hyperplasia effect on the endometrium. PMID- 17419679 TI - Differential binding of poly(ADP-Ribose) polymerase-1 and JunD/Fra2 accounts for RANKL-induced Tcirg1 gene expression during osteoclastogenesis. AB - We studied Tcirg1 gene expression on RANKL-induced osteoclastic differentiation of the mouse model RAW264.7 cells. We identified a mechanism involving PARP-1 inhibition release and JunD/Fra-2 binding, which is responsible for Tcirg1 gene upregulation. INTRODUCTION: The Tcirg1 gene encodes the a3 isoform of the V ATPase a subunit, which plays a critical role in the resorption activity of the osteoclast. Using serial deletion constructs of the Tcirg1 gene promoter, we performed a transcriptional study to identify factor(s) involved in the regulation of the RANKL-induced gene expression. MATERIALS AND METHODS: The promoter activity of serial-deletion fragments of the Tcirg1 gene promoter was monitored throughout the RAW264.7 cells differentiation process. We next performed sequence analysis, EMSA, UV cross-linking, qPCR, and gel supershift experiments to identify the factor(s) interacting with the promoter. RESULTS: A deletion of the -1297-1244 region led to the disappearance of the RANKL-induced promoter activity. EMSA experiments showed the binding of two factors that undergo differential binding on RANKL treatment. Supershift experiments led us to identify the dimer JunD/Fra-2 as the binding activity associated with the -1297/ 1268 Tcirg1 gene promoter sequence in response to RANKL. Moreover, we observed poly(ADP-ribose) polymerase-1 (PARP-1) binding to an adjacent site (-1270/-1256), and this interaction was disrupted after RANKL treatment. CONCLUSIONS: We provide data that identify junD proto-oncogene (JunD) and Fos-related antigen 2 (Fra-2) as the activator protein-1 (AP-1) factors responsible for the RANKL-induced upregulation of the mouse Tcirg1 gene expression. Moreover, we identified another binding site for PARP-1 that might account for the repression of Tcirg1 gene expression in pre-osteoclastic cells. PMID- 17419680 TI - Polyethylene particle-induced bone resorption in alpha-calcitonin gene-related peptide-deficient mice. AB - This study investigates the impact of alpha-CGRP on bone metabolism after implantation of polyethylene particles. alpha-CGRP knockout mice showed less osteolysis compared with wildtype mice. The local neurogenic microenvironment might be a crucial factor in particle-induced osteolysis. INTRODUCTION: Periprosthetic osteolysis is the major reason for aseptic loosening in joint arthroplasty. This study aimed to investigate the potential impact of alpha calcitonin gene-related peptide (alpha-CGRP) deficiency on bone metabolism under conditions of polyethylene particle-induced osteolysis. MATERIALS AND METHODS: We used the murine calvarial osteolysis model based on polyethylene particles in 14 C57BL 6 mice and 14 alpha-CGRP-deficient mice divided into four groups of 7 mice each. Groups 1 (C57BL/J 6) and 3 (alpha-CGRP knockout) received sham surgery, and groups 2 (C57BL/J 6) and 4 (alpha-CGRP knockout) were treated with polyethylene particles. Qualitative and quantitative 3D analyses were performed using microCT. In addition, bone resorption was measured within the midline suture by histological examination. The number of osteoclasts was determined by counting the TRACP(+) cells. Calvarial bone was tested for RANKL expression by RT-PCR and immunocytochemistry. RESULTS: Bone resorption was significantly reduced in alpha CGRP-deficient mice compared with their corresponding wildtype C57BL 6 mice as confirmed by histomorphometric data (p < 0.001) and microCT (p < 0.01). Osteoclast numbers were significantly reduced in group 3 and the particle subgroup compared with group 1 (p < 0.001). We observed a >3-fold increase of basal RANKL mRNA levels within group 1 compared with group 3. Additional low RANKL immunochemistry staining was noted in groups 3 and 4. CONCLUSIONS: In conclusion, alpha-CGRP knockout mice did not show the expected extended osteolysis compared with wildtype mice expressing alpha-CGRP. One of the most reasonable explanations for the observed decrease in osteolysis could be linked to the osteoprotegerin (OPG)/RANK/RANKL system in alpha-CGRP-deficient animals. As a consequence, the fine tuning of osteoclasts mediating resorption in alpha CGRP-null mice may be deregulated. PMID- 17419681 TI - Inflammatory markers and incident fracture risk in older men and women: the Health Aging and Body Composition Study. AB - The inflammation of aging hypothesis purports that aging is the accumulation of damage, which results, in part, from chronic activation of inflammation process. We tested this hypothesis in relationship to fractures in 2985 men and women enrolled in the Health ABC study. Results showed that subjects with the greatest number of inflammatory markers have the highest risk of fracture. INTRODUCTION: Cytokines play major roles in regulating bone remodeling in the bone microenvironment, but their relationship to fractures is uncertain. MATERIALS AND METHODS: The study population includes 2985 well-functioning white and black women and men (42%, black; 51%, women) 70-79 yr of age enrolled in the Health Aging and Body Composition Study. Inflammatory markers were measured in frozen serum using standardized assays. We measured interleukin (IL-6), TNFalpha, C reactive protein (CRP), and soluble receptors (IL-2 sR, IL-6 sR, TNF sR1and TNF sR2).Cytokine-soluble receptors were measured in a subset (n = 1430). Total hip BMD was measured by DXA. During 5.8 +/- 1.6 yr of 95% complete follow-up, incident fractures were confirmed in 268 subjects. The risk of fracture was compared among subjects with the highest inflammatory markers (quartile 4) versus lower levels (quartiles 1, 2, and 3) using proportional hazard models. RESULTS AND CONCLUSIONS: Subjects who fractured were more likely to be white and female. Baseline markers of inflammation were higher among subjects who subsequently experienced an incident fracture. In multivariate models, the relative risk of fracture (95% CIs) for subjects with the highest inflammatory markers (quartile 4) compared with those with lower inflammatory markers (quartiles 1, 2, and 3) was 1.34 (0.99, 1.82) for CRP; 1.28 (0.95-1.74) for IL-6; 1.28 (0.97-1.70) for TNFalpha; 1.52 (1.04-2.21) for IL-2 sR; 1.33 (0.90-1.96) for IL-6 sR; 1.73 (1.18 2.55) for TNF sR1 and 1.48 (1.01-2.20) for TNF sR2. In subjects with three or more (out of seven) high inflammatory markers, the relative risk of fracture was 2.65 (1.44-4.89) in comparison with subjects with no elevated markers. (p trend = 0.001). We conclude that elevated inflammatory markers are prognostic for fractures, extending the inflammation hypothesis of aging to osteoporotic fractures. PMID- 17419682 TI - Evolutionary tinkering: birth of a novel chloroplast protein. AB - The term 'evolutionary tinkering' refers to evolutionary innovation by recombination of functional units, and includes the creation of novel proteins from pre-existing modules. A novel instance of evolutionary tinkering was recently discovered in the flowering plant genus Nicotiana: the conversion of a nuclear transcription factor into the plastid-resident protein WIN4 (wound induced clone 4) involved in environmental stress responses. In this issue of the Biochemical Journal, Kodama and Sano now show that two steps are necessary for the establishment of the novel plastid protein: the acquisition of an internal translation initiation site and the use of multiple transcription starts to produce short mRNA variants that encode the plastid-targeted protein form. PMID- 17419683 TI - Pathways of signal transduction employed by vertebrate Hedgehogs. AB - Signalling by Hh (Hedgehog) proteins is among the most actively studied receptor mediated phenomena relevant to development and post-embryonic homoeostatic events. The impact of signalling by the Hh proteins is profound, and work pertaining to the presentation of these proteins and the pathways engaged by them continues to yield unique insights into basic aspects of morphogenic signalling. We review here the mechanisms of signalling relevant to the actions of Hh proteins in vertebrates. We emphasize findings within the past several years on the recognition of, in particular, Sonic hedgehog by target cells, pathways of transduction employed by the seven-pass transmembrane protein Smoothened and end points of action, as manifest in the regulation of the Gli transcription factors. Topics of extended interest are those regarding the employment of heterotrimeric G-proteins and G-protein-coupled receptor kinases by Smoothened. We also address the pathways, insofar as known, linking Smoothened to the expression and stability of Gli1, Gli2 and Gli3. The mechanisms by which Hh proteins signal have few, if any, parallels. It is becoming clear in vertebrates, however, that several facets of signalling are shared in common with other venues of signalling. The challenge in understanding both the actions of Hh proteins and the overlapping forms of regulation will be in understanding, in molecular terms, both common and divergent signalling events. PMID- 17419684 TI - What is the role of non-invasive measurements of atherosclerosis in individual cardiovascular risk prediction? AB - Primary prevention of CVD (cardiovascular disease) is mainly based on the assessment of individual cardiovascular risk factors. However, often, only the most important (conventional) cardiovascular risk factors are determined, and every level of risk factor exposure is associated with a substantial variation in the amount of atherosclerosis. Measuring the effect of risk factor exposure over time directly in the vessel might (partially) overcome these shortcomings. Several non-invasive imaging techniques have the potential to accomplish this, each of these techniques focusing on a different stage of the atherosclerotic process. In this review, we aim to define the current role of various of these non-invasive measurements of atherosclerosis in individual cardiovascular risk prediction, taking into account the most recent insights about validity and reproducibility of these techniques and the results of recent prospective outcome trials. We conclude that, although the clinical application of FMD (flow-mediated dilation) and PWA (pulse wave analysis) in individual cardiovascular risk prediction seems far away, there may be a role for PWV (pulse wave velocity) and IMT (intima-media thickness) measurements in the near future. PMID- 17419685 TI - Medroxyprogesterone and conjugated oestrogen are equivalent for hot flushes: a 1 year randomized double-blind trial following premenopausal ovariectomy. AB - Oestrogen therapy is the gold standard treatment for hot flushes/night sweats, but it and oestrogen/progestin are not suitable for all women. MPA (medroxyprogesterone acetate) reduces hot flushes, but its effectiveness compared with oestrogen is unknown. In the present study, oral oestrogen [CEE (conjugated equine oestrogen)] and MPA were compared for their effects on hot flushes in a planned analysis of a secondary outcome for a 1-year randomized double-blind parallel group controlled trial in an urban academic medical centre. Participants were healthy menstruating women prior to hysterectomy/ovariectomy for benign disease. A total of 41 women {age, 45 (5) years [value is mean (S.D.)]} were enrolled; 38 women were included in this analysis of daily identical capsules containing CEE (0.6 mg/day) or MPA (10 mg/day). Demographic variables did not differ at baseline. Daily data provided the number of night and day flushes compared by group. The vasomotor symptom day-to-day intensity change was assessed by therapy assignment. Hot flushes/night sweats were well controlled in both groups, one occurred on average every third day and every fourth night. Mean/day daytime occurrences were 0.363 and 0.187 with CEE and MPA respectively, but were not significantly different (P=0.156). Night sweats also did not differ significantly (P=0.766). Therapies were statistically equivalent (within one event/24 h) in the control of vasomotor symptoms. Day-to-day hot flush intensity decreased with MPA and tended to remain stable with CEE (P<0.001). In conclusion, this analysis demonstrates that MPA and CEE are equivalent and effective in the control of the number of hot flushes/night sweats immediately following premenopausal ovariectomy. PMID- 17419686 TI - Sublingual immunotherapy for asthma: need for high quality meta-analyses to prove the concept. PMID- 17419690 TI - Incretory renal functions--Tigerstedt, renin and its neglected antagonist medullipin. PMID- 17419691 TI - Methyl aminolaevulinate-photodynamic therapy: a review of clinical trials in the treatment of actinic keratoses and nonmelanoma skin cancer. AB - Methyl aminolaevulinate-photodynamic therapy (MAL-PDT) has advanced the management of nonmelanoma skin cancer (NMSC), providing a treatment option for actinic keratosis (AK), basal cell carcinoma [both superficial (sBCC) and nodular (nBCC)] and Bowen's disease, with good clinical outcomes, low recurrence rates and enhanced cosmetic acceptability. Excellent results have been reported, with complete responses (CRs) in AK ranging from 69% to 93% at 3 months; CRs in Bowen's disease are 93% at 3 months and 68% at 24 months. In sBCC, CRs range from 85% to 93% at 3 months and are comparable with cryosurgery up to 60 months (75% vs. 74%). In nBCC, CRs range from 75-82% at 3 months to 77% at 60 months. MAL-PDT specifically targets diseased cells, leaving healthy tissue unharmed. This noninvasive treatment option is associated with minimal risk of scarring. Moreover, systemic uptake of MAL is negligible and the local phototoxic reactions that often occur during treatment rapidly heal to produce excellent cosmetic results. The side-effects of therapy, which are predominantly local phototoxic effects (burning, stinging and prickling sensations), are of mild-to-moderate intensity, of short duration and easily managed. Overall, the efficacy and low risk of side-effects afforded by this therapy have resulted in high patient preference in clinical trials. The current evidence base for MAL-PDT in the treatment of AK and NMSC is reviewed in this article. PMID- 17419692 TI - Gene expression profiling analysis of solar lentigo in relation to immunohistochemical characteristics. AB - BACKGROUND: Solar lentigo appears as dark brown spots that occur on sun-exposed areas and is considered to be a hallmark of aged skin. Although considerable knowledge about acute pigmentation has recently been accumulated, little is yet known about the mechanisms underlying chronic- and delayed-type hyperpigmentation, such as solar lentigo. OBJECTIVES: To clarify further the mechanisms underlying the development of solar lentigo, we carried out gene expression analysis in skin biopsy specimens obtained from human solar lentigines using DNA microarray analysis. METHODS: Two pairs of skin specimens were obtained from solar lentigo and adjacent sun-exposed normal skin, as well as normal skin on the buttocks of 16 volunteers aged 40-55 years. One set of specimens was frozen and RNA was extracted for microarray and the other set was prepared for histological sections and analysed by antibodies and probes. RESULTS: Sixty-five genes were upregulated more than 1.8-fold in solar lentigo compared with adjacent control skin and seven melanocyte-related genes were included. Compared with sun protected skin, many inflammation-related genes were upregulated in solar lentigo, and compared with sun-exposed control skin, upregulation of genes related to fatty-acid metabolism was apparent in solar lentigo. Moreover, we found downregulation of cornified envelope-related genes, which suggests suppression of cornification in the epidermis in solar lentigo. Immunohistochemically, larger numbers of TRP1-positive cells were found in the basal layer of solar lentigo than in normal skin. Fatty acid-related genes were highly expressed in the epidermis as detected by in situ hybridization, and they were much more prominent in the lesional skin of solar lentigo. However, cycling epidermal cells detectable with Ki67 antibody were fewer in the lesional skin of solar lentigo. Expression of filaggrin and involucrin was decreased in the lesional skin, where the number of cell layers of the stratum corneum was significantly higher than in normal skin. CONCLUSIONS: The results of the present microarray analysis of solar lentigo, demonstrating upregulation of genes related to inflammation, fatty-acid metabolism and melanocytes and downregulation of cornified envelope-related genes, suggest that solar lentigo is induced by the mutagenic effect of repeated ultraviolet light exposures in the past, leading to the characteristic enhancement of melanin production, together with decreased proliferation and differentiation of lesional keratinocytes on the background of chronic inflammation. PMID- 17419693 TI - Sclerodermatous graft-versus-host disease: clinical spectrum and therapeutic challenges. AB - Sclerodermatous graft-versus-host disease (GVHD) is a rare complication of bone marrow transplantation. While GVHD is often associated with the beneficial graft vs. tumour effect, it also contributes towards significant morbidity and mortality. No reliably effective treatment has yet been established. We present 10 patients with haematological malignancies who underwent an allogeneic stem cell transplant and developed sclerodermatous GVHD. Donor lymphocyte infusion administered for relapse or reducing donor T-cell chimerism was a known trigger for sclerodermatous GVHD in four of the patients. Treatment with immunosuppressants, psoralen plus ultraviolet A (PUVA) and extracorporeal photopheresis has been largely unsuccessful in their management. Intensive immunosuppression including the use of anti-CD20 monoclonal antibody may have contributed to relapse of leukaemia in one patient 10 years after her transplant. Sclerodermatous GVHD may occur without a preceding lichenoid stage. Clinical heterogeneity is common, although sclerodermatous GVHD has a predilection for the limbs. Treatment options are largely unsatisfactory if conventional immunosuppression fails. PUVA may give some symptomatic benefit and extracorporeal photopheresis seems to be less efficacious than previously published work suggests. PMID- 17419694 TI - Should urologists be spending more time on the golf course? PMID- 17419695 TI - Case number and the financial impact of circumcision in reducing prostate cancer. PMID- 17419696 TI - Stopping smoking might reduce tumour recurrence in nonmuscle-invasive bladder cancer. AB - OBJECTIVE: To evaluate effects of stopping smoking on the outcome of nonmuscle invasive bladder cancer, as cigarette smoking is a risk factor for bladder cancer and little is known about whether stopping smoking reduces the risk of recurrence or progression. PATIENTS AND METHODS: Between January 1997 and July 2005, 297 men with primary nonmuscle-invasive bladder cancer were treated with transurethral resection (TUR); their smoking status before and after the diagnosis of bladder cancer was obtained by a post hoc questionnaire and interview. 'Quitters' were those who ceased smoking within a year before and 3 months after the diagnosis. Ex-smokers were those who ceased smoking more than a year before diagnosis. Several pathological and clinical variables were compared, with all statistical comparisons being two-sided. RESULTS: In all, 265 patients completed the questionnaire, including 64 non-smokers, 64 ex-smokers, 59 quitters, and 78 continued smokers. The median follow-up was 38 months. There were no significant differences in the strata of stage, grade, tumour multiplicity, intravesical therapy, or median follow-up duration between the four patient groups. The respective 3-year recurrence-free survival of continued smokers, non-smokers, ex smokers and quitters was 45%, 57%, 62% and 70%. By multivariate analysis, high grade, T1-stage, multiple tumours and continued smoking were significant independent predictors for a shorter recurrence-free survival. Quitters had a lower risk of recurrence than did either continued smokers or non-smokers, but had a similar risk to ex-smokers. CONCLUSION: Stopping smoking might be associated with a lower recurrence rate for patients with nonmuscle-invasive bladder cancer. PMID- 17419697 TI - Peri-operative complications of holmium laser enucleation of the prostate: experience in the first 280 patients, and a review of literature. AB - OBJECTIVE: To evaluate, in a prospective study, the complications in 280 patients undergoing holmium laser enucleation of the prostate (HoLEP) at our institution, and to review previous reports to determine the overall incidence and types of various complications, and analyse their causes and means of prevention. PATIENTS AND METHODS: We analysed the patients' demographic, peri-operative and follow-up data, and the complications during and after surgery. RESULTS: HoLEP was completed successfully in 268 patients (95.7%); eight required conversion to transurethral resection of the prostate (TURP) during the initial experience. The morcellation device and laser malfunctioned in two patients each. A blood transfusion was required during HoLEP in one patient; other complications included capsular perforation (9.6%), superficial bladder mucosal injury (3.9%) and ureteric orifice injury (2.1%). A blood transfusion was needed after HoLEP in 1.4% of patients and cystoscopy with clot evacuation in 0.7%. Transient urinary incontinence was the commonest complication after HoLEP, in 10.7% of patients, but recovered spontaneously in all except two (0.7%). Other rare complications were re-catheterization (3.9%), urinary tract infection (3.2%), epididymitis (0.7%), meatal and submeatal stenosis (2.5%), bulbar urethral stricture (2.1%), bladder neck contracture (0.35%) and myocardial infarction (0.35%). CONCLUSIONS: There was a low incidence of complications with HoLEP; most were minor and easily managed. Our results are comparable with those published previously, and establish HoLEP as safe and reproducible procedure. While gaining experience, HoLEP can be converted to TURP with no harm to the patient. PMID- 17419698 TI - Interstitial laser coagulation in the management of lower urinary tract symptoms suggestive of bladder outlet obstruction from benign prostatic hyperplasia: long term follow-up. AB - OBJECTIVE: To evaluate the long-term effects, including durability, of interstitial laser coagulation (ILC) in patients with symptomatic benign prostatic hyperplasia. PATIENTS AND METHODS: In all, 49 men (median age 68 years, range 52-80) were treated with ILC; 22 men were assessed for a median (range) of 54 (46-61) months. Information on the timing and type of re-treatment were collected for the remaining patients. RESULTS: The median (quartiles) International Prostate Symptom Score decreased from 22 (19-28) at baseline to 13 (5-21), a decrease of 41%. The peak urinary flow increased by 20% to 10.2 (8.7 12.9) mL/s. Twenty-two patients were re-treated (by any type) after ILC, giving a re-treatment rate of 50%. CONCLUSION: There were moderate effects on voiding variables and a high re-treatment rate during the long-term follow-up after ILC. It seems reasonable that the use of ILC is restricted to selected, high-risk patients. PMID- 17419699 TI - Urologists: be aware of significant risks to stopping anticoagulants in patients with drug-eluting coronary stents. PMID- 17419700 TI - Basal cell carcinoma of the prostate: current concepts. PMID- 17419701 TI - Laparoscopy in paediatric urology: present status. AB - The spectrum of laparoscopic surgery in children has developed dramatically; what was initially used as a diagnostic method to identify an impalpable testis is now commonly used for complex reconstructive procedures such as pyeloplasty. Laparoscopic orchidopexy and nephrectomy are well established and are used at many centres. Laparoscopic partial nephrectomy, adrenalectomy and dismembered pyeloplasty series have reported shorter hospital stays and operative times that are comparable with that of open techniques, and/or decreasing with experience. The initial experiences with laparoscopic ureteric re-implantation and laparoscopically assisted bladder reconstructive surgery are reported, with encouraging results for feasibility, hospital stay, and cosmetic outcome. PMID- 17419702 TI - The stress response in laparoscopic urological surgery. PMID- 17419703 TI - The predictive value of preoperative pressure-flow studies in the resolution of detrusor overactivity and overactive bladder after tension-free vaginal tape insertion. AB - OBJECTIVE: To determine whether preoperative pressure-flow studies (PFS) predict the resolution of detrusor overactivity (DO) and overactive bladder (OAB) symptoms after a tension-free vaginal tape procedure (TVT). PATIENTS AND METHODS: Thirty-five consecutive women with mixed DO and urodynamic stress incontinence (USI) undergoing a TVT had PFS before and afterward reviewed, and the results compared. RESULTS: There was resolution of OAB symptoms in 51%; the persistence of OAB symptoms was predicted by a significant decrease (20.0 to 14.0 mL/s) in the maximum flow rate after the TVT (P = 0.027) and a significant increase in the detrusor pressure at maximum flow after the TVT (P = 0.04). DO was absent on cystometry in 46% of women after the TVT. Women with persistent DO on cystometry had a significantly lower (P = 0.02) maximum flow rate before the TVT (mean 19.3 mL/s) than those with no persistent DO (mean 26.9 mL/s). This finding persisted when flow rates were corrected for voided volume (P = 0.04). Before and after TVT there were no significant differences between the groups in voiding time and acceleration of flow. USI was objectively cured in 92% of the women. CONCLUSIONS: Women whose maximum flow rate decreases significantly after the TVT are more likely to have persistent OAB symptoms. The urinary flow rate before the TVT was significantly higher in women with an objective cure of DO after TVT than in women with persistent DO. These findings support an obstructive cause in women in whom DO does not resolve. PMID- 17419704 TI - Europa uomo. PMID- 17419705 TI - Association of vitamin D receptor (Fok-I) polymorphism with the clinical presentation of calcium urolithiasis. AB - OBJECTIVE: To investigate the effect of the vitamin D receptor (VDR) FokI polymorphism on the clinical presentation of calcium urolithiasis, as a FokI polymorphism in the VDR gene was recently reported to be associated with calcium metabolism disorders. PATIENTS, SUBJECTS AND METHODS: In all, 235 patients with calcium urolithiasis and 231 age- and sex-matched healthy controls were recruited from Kaohsiung Medical University Hospital between June 2003 and February 2005. Clinical information on the age at first onset, stone episodes, stone severity and presence of family history were collected from patients with stones. Any VDR FokI polymorphism was detected using polymerase chain reaction-based restriction analysis. RESULTS: The frequency of VDR FokI genotypes between the patients and the healthy controls was not significantly different. However, among patients, those with the FF genotype had a significantly higher risk of having more stone episodes (adjusted odds ratio 2.15, 95% confidence interval 1.02-4.54, P = 0.044) and were younger at the first onset (3.23, 1.08-9.63, P = 0.036) than those with the ff genotype. CONCLUSION: The VDR FokI polymorphism might be important in the clinical presentation of patients with calcium urolithiasis, especially for the frequency of stone episodes and age at first onset, although it is not associated with the formation of stones. PMID- 17419706 TI - Proteomic analysis of rat penile tissue in a model of erectile dysfunction after radical prostatectomy. AB - OBJECTIVE: To identify differential protein expression in penile tissue in a rat model of erectile dysfunction (ED) at an early stage after bilateral cavernosal nerve (CN) neurectomy, using proteomic techniques. MATERIALS AND METHODS: Twelve male adult Sprague-Dawley rats were randomly divided into two equal groups, one having bilateral CN resection and one a control group. The penises were harvested 7 days after CN resection. Total protein was separated into >1250 protein spots by two-dimensional electrophoresis using pH 3-10 nonlinear immobilized pH gradient strips. Differential expression of proteins was analysed using matrix assisted laser desorption/ionization time-of-flight mass spectrometry and database searching. RESULTS: Thirty-two proteins were significantly changed in the denervated penis, of which 25 (including nine up-regulated and 16 down regulated) with cytoskeletal functions, and pathophysiological functions related to energy metabolism and oxidative stress, were identified. Examples include transgelin, creatine kinase B, annexin-1 and galactin-7. CONCLUSIONS: The expression of several important proteins participating in pathophysiological processes of penile tissue are changed early after bilateral CN neurectomy. These changes might give new insights into the cellular and molecular mechanisms involved in neurogenic ED development, and indicate potential therapeutic targets. PMID- 17419707 TI - A review of targeted screening for prostate cancer: introducing the IMPACT study. PMID- 17419708 TI - Sonodynamic therapy of cancer using a novel porphyrin derivative, DCPH-P-Na(I), which is devoid of photosensitivity. AB - To improve the efficacy of sonodynamic therapy of cancer using photosensitizers, we developed a novel porphyrin derivative designated DCPH-P-Na(I) and investigated its photochemical characteristics and sonotoxicity on tumor cells. DCPH-P-Na(I) exhibited a minimum fluorescent emission by excitation with light, compared with a strong emission from ATX-70, which is known to reveal both photo- and sonotoxicity. According to this observation, when human tumor cells were exposed to light in the presence of DCPH-P-Na(I) in vitro, the least phototoxicity was observed, in contrast to the strong phototoxicity of ATX-70. However, DCPH-P-Na(I) exhibited a potent sonotoxicity on tumor cells by irradiation with ultrasound in vitro. This sonotoxicity was reduced by the addition of L-histidine, but not D-mannitol, thus suggesting that singlet oxygen may be responsible for the sonotoxicity of DCPH-P-Na(I). DCPH-P-Na(I) demonstrated significant sonotoxicity against a variety of cancer cell lines derived from different tissues. In addition, in a mouse xenograft model, a potent growth inhibition of the tumor was observed using sonication after the administration of DCPH-P-Na(I) to the mouse. These results suggest that sonodynamic therapy with DCPH-P-Na(I) may therefore be a useful clinical treatment for cancers located deep in the human body without inducing skin sensitivity, which tends to be a major side-effect of photosensitizers. PMID- 17419709 TI - Signal transduction mediated by endostatin directly modulates cellular function of lung cancer cells in vitro. AB - Endostatin (ED) is a carboxyl-terminal fragment of collagen XVIII with strong antiangiogenic activity. ED has been considered as a highly specific inhibitor of endothelial cell proliferation and migration through interaction with its receptor on the surface of endothelial cells. Recently, direct antitumor effects of ED in colon cancer cells and head and neck squamous cell carcinoma cells has been reported. However, its effect on lung cancer cells has not been clarified. The purpose of the present study was to determine the effect of ED on in vitro lung cancer cell function and to identify its receptor on lung cancer cells. We revealed that alpha5 integrin is capable of being a functional ED receptor among several integrins that are expressed on murine lung cancer (Lewis lung cancer [LLC]) cells. We further demonstrated that the ED-integrin interaction modulates various in vitro biological functions of LLC cells as we revealed that immobilized ED helps in LLC cell adhesion and migration in an integrin-dependent manner. Furthermore, ED inhibited LLC cell proliferation and induced apoptosis. Interestingly, ED did not demonstrate any antiproliferative activity against the other murine lung cancer cell line, KLN205, that lacks alpha5 integrin but binds to immobilized ED through the beta1 integrin. In addition, the binding of ED to alpha5 integrin on LLC cells induced phosphorylation of focal adhesion kinase. Taken together, these results suggest that the interaction between ED and alpha5 integrin may play an important role in lung cancer cell function. PMID- 17419710 TI - Serum albumin-associated peptides of patients with uterine endometrial cancer. AB - The incidence of endometrial cancer is predicted to increase in developed countries. Because of the relatively high incidence of complications and low diagnostic sensitivity associated with endometrial tissue sampling, there is an urgent need for the development of a safe and non-invasive diagnostic method. The proteomic spectrum of albumin-associated peptides was obtained from a total of 125 serum samples (92 from endometrial cancer patients and 33 from controls) by matrix-assisted laser desorption/ionization hybrid quadrupole time-of-flight mass spectrometry, and the candidate markers were selected by the Mann-Whitney U-test and receiver operator characteristics analysis. We selected three mass peaks at 4769, 6254 and 11 792 m/z from a total of 507 peaks as distinguishing cancer patients from controls (P < 0.00001 and area under curve of over 0.8). When the cut-off points were defined as the averages of the values in the controls + 2 SD, the combination of the three peptides detected endometrial cancer with a sensitivity of 65.2% (60/92). Even stage I early endometrial cancers were detected with a sensitivity of 60.3% (38/63). Unfortunately, the three peptides were also detected in 44.6% (33/74) of myoma uteri patients, indicating that they are not specific to endometrial cancer. Although a large-scale study is necessary to confirm the clinical significance of the peptide biomarkers identified in this study, direct profiling of serum-albumin-bound peptides by high-resolution mass spectrometry was proven to have potential as a means of identifying biomarkers for a variety of diseases. PMID- 17419711 TI - Heparanase gene haplotype (CGC) is associated with stage of disease in patients with ovarian carcinoma. AB - Heparanase (HSPE-1) and vascular endothelial growth factor (VEGF), proangiogenic growth factors, play important roles in the metastatic biology of ovarian cancer. The aim of the present study was to test for association between single nucleotide polymorphisms (SNPs) in HSPE-1 and VEGF and outcome in ovarian cancer. A mutational analysis was performed on the coding sequence of the HSPE-1 gene to define high-frequency SNPs. HSPE-1 polymorphisms, together with two SNPs in the VEGF gene, were studied in 136 patients with ovarian cancer. Patients were categorized into two groups, those with FIGO stages 1 and 2 (group 1) and those with stages 3 and 4 (group 2). We identified 10 polymorphisms in the HSPE-1 gene, those in introns 2, 3 and 5b, and exons 8, 13a and 13b occurring at a minor allele frequency of >/=10%. There was an increase in frequency of those individuals with a genotype that carried at least one copy of the intron 2 (C), exon 8 (G), exon 13a (C) haplotype (CGC) in group 2. Specifically there were 24% with this haplotype in group 2 versus 5% in group 1 (P = 0.0184, odds ratio 5.986, 95% confidence interval 1.340-26.752). Most of this association was captured by the intron 2 genotype, where carriage of the C allele was associated with stage (P = 0.0148, odds ratio 6.524, 95% confidence interval 1.401-27.921). There was no association between VEGF SNPs and stage of disease. The CGC HSPE-1 haplotype associates with stage in ovarian cancer. This haplotype may affect splicing of the HSPE-1 gene, as in silico it alters the presence of a splicing motif. PMID- 17419712 TI - Anti-beta1-adrenergic receptor autoantibodies in patients with chronic Chagas heart disease. AB - Chronic Chagas heart disease (cChHD), a chronic manifestation of the Trypanosoma cruzi infection, is characterized by high antibody levels against the C-terminal region of the ribosomal P proteins (i.e. peptide R13, EEEDDDMGFGLFD) which bears similarity with the second extracellular loop of beta1-adrenergic receptor (beta1 AR, peptide H26R HWWRAESDEARRCYNDPKCCDFVTNR). Because it has not been demonstrated clearly that IgGs from cChHD patients bind to native human beta1-AR, the aim of this study was to investigate further the physical interaction between cChHD IgGs and the human beta1-AR. Immunofluorescence assays demonstrated the binding of these antibodies to the receptor expressed on stably transfected cells, together with a beta1-AR agonist-like effect. In addition, immunoadsorption of the serum samples from cChHD patients with a commercially available matrix, containing peptides representing the first and the second extracellular loop of the beta1-AR, completely abolished reactivity against the H26R peptide and the physiological response to the receptor. The follow-up of this specificity after in vitro immunoadsorption procedures suggests that this treatment might be used to diminish significantly the serum levels of anti-beta1 AR antibodies in patients with Chagas heart disease. PMID- 17419713 TI - Role of plant stomata in bacterial invasion. AB - Stomata are microscopic pores in the epidermis of the aerial parts of terrestrial plants. These pores are essential for photosynthesis, as they allow CO(2) to diffuse into the plant. The size of the stomatal pore changes in response to environmental conditions, such as light intensity, air humidity and CO(2) concentrations, as part of the plant's adaptation to maximize photosynthetic efficiency and, at the same time, to minimize water loss. Historically, stomata have been considered as passive portal of entry for plant pathogenic bacteria. However, recent studies suggest that stomata can play an active role in restricting bacterial invasion as part of the plant innate immune system. Some plant pathogens have evolved specific virulence factors to overcome stomata-based defence. Interestingly, many bacterial disease outbreaks require high humidity, rain, or frost damage, which could promote stomatal opening and/or bypass stomatal defence by creating wounds as alternative entry sites. Further studies on microbial and environmental regulation of stomata-based defence should fill gaps in our understanding of bacterial pathogenesis, disease epidemiology and phyllosphere microbiology. PMID- 17419714 TI - Immune activation suppresses initiation of lytic Epstein-Barr virus infection. AB - Primary infection with Epstein-Barr virus (EBV) is asymptomatic in children with immature immune systems but may manifest as infectious mononucleosis, a vigorous immune activation, in adolescents or adults with mature immune systems. Infectious mononucleosis and chronic immune activation are linked to increased risk for EBV-associated lymphoma. Here we show that EBV initiates progressive lytic infection by expression of BZLF-1 and the late lytic genes gp85 and gp350/220 in cord blood mononuclear cells (CBMC) but not in peripheral blood mononuclear cells (PBMC) from EBV-naive adults after EBV infection ex vivo. Lower levels of proinflammatory cytokines in CBMC, used to model a state of minimal immune activation and immature immunity, than in PBMC were associated with lytic EBV infection. Triggering the innate immunity specifically via Toll-like receptor 9 of B cells substantially suppressed BZLF-1 mRNA expression in acute EBV infection ex vivo and in anti-IgG-stimulated chronically latently EBV-infected Akata Burkitt lymphoma cells. This was mediated in part by IL-12 and IFN-gamma. These results identify immune activation as critical factor for the suppression of initiation of lytic EBV infection. We hypothesize that immune activation contributes to EBV-associated lymphomagenesis by suppressing lytic EBV and in turn promotes latent EBV with transformation potential. PMID- 17419715 TI - Activation of the virus-induced IKK/NF-kappaB signalling axis is critical for the replication of human cytomegalovirus in quiescent cells. AB - Activation of the IKK/NF-kappaB signalling pathway is a hallmark of human cytomegalovirus (HCMV) infection. However, its role in regulating major immediate early promoter (MIEP)-dependent transcription and HCMV replication remains controversial. This study uses a combination of genetic approaches to investigate the effects of cell culture conditions on the importance of virus-induced NF kappaB activation during the infection of endothelial cells or fibroblasts. Adenoviral-mediated expression of a dominant-negative mutant of IKK2 kinase (dnIKK2) in human umbilical vein endothelial cells resulted in a strong reduction of IkappaBalpha degradation and NF-kappaB activation following infection with an HCMV clinical isolate. Viral replication was impaired in dnIKK2-expressing cells that were growth-arrested before infection, but not in replicating cells. The inhibitory effect of dnIKK2 was independent from the virus strain and the cell type used, because replication of the laboratory AD169 strain was impaired as well in dnIKK2-expressing quiescent fibroblasts. Moreover, progressive disruption of NF-kappaB response elements within the MIEP in recombinant HCMV viruses derived from the clinical isolate prevented their replication in quiescent cells but not in actively growing cells. These results demonstrate an essential role of virus-induced IKK/NF-kappaB activity to trigger both viral IE gene expression and productive replication in quiescent cells. PMID- 17419716 TI - Lipopolysaccharides from atherosclerosis-associated bacteria antagonize TLR4, induce formation of TLR2/1/CD36 complexes in lipid rafts and trigger TLR2-induced inflammatory responses in human vascular endothelial cells. AB - Infection with bacteria such as Chlamydia pneumonia, Helicobacter pylori or Porphyromonas gingivalis may be triggering the secretion of inflammatory cytokines that leads to atherogenesis. The mechanisms by which the innate immune recognition of these pathogens could lead to atherosclerosis remain unclear. In this study, using human vascular endothelial cells or HEK-293 cells engineered to express pattern-recognition receptors (PRRs), we set out to determine Toll-like receptors (TLRs) and functionally associated PRRs involved in the innate recognition of and response to lipopolysaccharide (LPS) from H. pylori or P. gingivalis. Using siRNA interference or recombinant expression of cooperating PRRs, we show that H. pylori and P. gingivalis LPS-induced cell activation is mediated through TLR2. Human vascular endothelial cell activation was found to be lipid raft-dependent and to require the formation of heterotypic receptor complexes comprising of TLR2, TLR1, CD36 and CD11b/CD18. In addition, we report that LPS from these bacterial strains are able to antagonize TLR4. This antagonistic activity of H. pylori or P. gingivalis LPS, as well as their TLR2 activation capability may be associated with their ability to contribute to atherosclerosis. PMID- 17419717 TI - Entry of the bacterial pathogen Listeria monocytogenes into mammalian cells. AB - The bacterial pathogen Listeria monocytogenes causes food-borne illnesses leading to meningitis or abortion. Listeria provokes its internalization ('entry') into mammalian cells that are normally non-phagocytic, such as intestinal epithelial cells and hepatocytes. Entry provides access to a nutrient-rich cytosol and allows translocation across anatomical barriers. Here I discuss the two major internalization pathways used by Listeria. These pathways are initiated by binding of the bacterial surface proteins InlA or InlB to their respective host receptors, E-cadherin or Met. InlA mediates traversal of the intestinal barrier, whereas InlB promotes infection of the liver. At the cellular level, both InlA- and InlB-dependent entry require host signalling that promotes cytoskeletal rearrangements and pathogen engulfment. However, many of the specific signalling proteins in the two entry routes differ. InlA-mediated uptake uses components of adherens junctions that are coupled to F-actin and myosin, whereas InlB-dependent entry involves cytosolic adaptors that bridge Met to regulators of F-actin, including phosphoinositide 3-kinase and activators of the Arp2/3 complex. Unexpectedly, entry directed by InlB also involves endocytic components. Future work on InlA and InlB will lead to a better understanding of virulence, and may also provide novel insights into the normal biological functions of E-cadherin and Met. PMID- 17419718 TI - The multiple mechanisms of Ca2+ signalling by listeriolysin O, the cholesterol dependent cytolysin of Listeria monocytogenes. AB - Cholesterol-dependent cytolysins (CDCs) represent a large family of conserved pore-forming toxins produced by several Gram-positive bacteria such as Listeria monocytogenes, Streptococcus pyrogenes and Bacillus anthracis. These toxins trigger a broad range of cellular responses that greatly influence pathogenesis. Using mast cells, we demonstrate that listeriolysin O (LLO), a prototype of CDCs produced by L. monocytogenes, triggers cellular responses such as degranulation and cytokine synthesis in a Ca(2+)-dependent manner. Ca(2+) signalling by LLO is due to Ca(2+) influx from extracellular milieu and release of from intracellular stores. We show that LLO-induced release of Ca(2+) from intracellular stores occurs via at least two mechanisms: (i) activation of intracellular Ca(2+) channels and (ii) a Ca(2+) channels independent mechanism. The former involves PLC-IP(3)R operated Ca(2+) channels activated via G-proteins and protein tyrosine kinases. For the latter, we propose a novel mechanism of intracellular Ca(2+) release involving injury of intracellular Ca(2+) stores such as the endoplasmic reticulum. In addition to Ca(2+) signalling, the discovery that LLO causes damage to an intracellular organelle provides a new perspective in our understanding of how CDCs affect target cells during infection by the respective bacterial pathogens. PMID- 17419719 TI - Intrinsic apoptotic pathways of gingival epithelial cells modulated by Porphyromonas gingivalis. AB - Porphyromonas gingivalis can inhibit chemically induced apoptosis in primary cultures of gingival epithelial cells through blocking activation of the effector caspase-3. The anti-apoptotic phenotype of P. gingivalis is conserved across strains and does not depend on the presence of fimbriae, as fimbriae-deficient mutants and a naturally occurring non-fimbriated strain were able to impede apoptosis. To dissect the survival pathways modulated by P. gingivalis, protein and gene expression of a number of components of apoptotic death pathways were investigated. P. gingivalis infection of epithelial cells resulted in the phosphorylation of JAK1 and Stat3. Quantitative real-time reverse transcription polymerase chain reaction showed that expression of Survivin and Stat3 itself, targets of activated Stat3, were elevated in P. gingivalis-infected cells. siRNA knockdown of JAK1, in combination with knockdown of Akt, abrogated the ability of P. gingivalis to block apoptosis. In contrast, cIAP-1 and cIAP-2 were not differentially regulated at either the protein or mRNA levels by P. gingivalis. One mechanism by which P. gingivalis can block apoptotic pathways in gingival epithelial cells therefore is through manipulation of the JAK/Stat pathway that controls the intrinsic mitochondrial cell death pathways. Induction of a pro survival phenotype may prevent programmed host cell death and aid survival of P. gingivalis within gingival epithelial cells. PMID- 17419720 TI - Dynamics and interactions of parvoviral NS1 protein in the nucleus. AB - Nuclear positioning and dynamic interactions of viral proteins with nuclear substructures play essential roles during infection with DNA viruses. Visualization of the intranuclear interactions and motility of the parvovirus replication protein (NS1) in living cells gives insight into specific parvovirus protein-cellular structure interactions. Confocal analysis of highly synchronized infected Norden Laboratory Feline Kidney cells showed accumulation of nuclear NS1 in discrete interchromosomal foci. NS1 fused with enhanced yellow fluorescence protein (NS1-EYFP) provided a marker in live cells for dynamics of NS1 traced by photobleaching techniques. Fluorescence Recovery after Photobleaching suggested that the NS1 protein is not freely diffusing but undergoes transient interactions with nuclear compartments. Fluorescence Loss in Photobleaching demonstrated for the first time the shuttling of a parvoviral protein between the nucleus and the cytoplasm as assayed with NS1-EYFP. Finally, time-lapse imaging of infected cells revealed that the intranuclear distribution of NS1-EYFP evolves dramatically starting from the formation of NS1 foci and proceeding to a homogenous distribution extending throughout the nucleus. PMID- 17419721 TI - Sleep-disordered breathing is related to an increased risk for type 2 diabetes in middle-aged men, but not in women--the FIN-D2D survey. AB - AIM: To determine the association between sleep-disordered breathing (SDB) and obesity, diabetes and glucose intolerance among middle-aged men and women in Finland. METHODS: A multicentre, population-based, cross-sectional survey in Finland. A total of 1396 men and 1500 women aged 45-74 years participated in the survey between 2004 and 2005. The study subjects underwent a health examination including an oral glucose tolerance test and filled a questionnaire describing their sleep habits. RESULTS: Middle-aged men with SDB had an increased prevalence of diabetes and abnormal glucose tolerance. These associations were not found among middle-aged women. After adjustments for age, body mass index, smoking and central nervous system-affecting medication, SDB was independently associated with diabetes and glucose intolerance in men, but not in women. CONCLUSION: Middle-aged men with SDB have an independent risk of type 2 diabetes. However, both diabetes and SDB exhibit a strong association with obesity and especially with central obesity, reflecting increased visceral fat. In clinical practice especially male patients with diabetes should always be asked about habitual snoring and about possible sleep apnoea. PMID- 17419722 TI - National Cholesterol Education Program and International Diabetes Federation definitions of metabolic syndrome in the prediction of diabetes. Results from the FIrenze-Bagno A Ripoli study. AB - BACKGROUND: The International Diabetes Federation (IDF) proposed to modify the diagnostic criteria for metabolic syndrome (MS) previously issued by the National Cholesterol Education Program (NCEP). Aim of the present investigation is to compare the predictive value for diabetes of NCEP and IDF definitions of MS in a large sample of predominantly Caucasian subjects. METHODS: A prospective observational study was performed on a cohort study (n = 3096) enrolled in a diabetes-screening programme, the FIrenze-Bagno A Ripoli study. All subjects with fasting glucose >126 mg/dl and/or post-load glucose > or =200 mg/dl (5.7%) were excluded from the present analysis. Follow-up of each subject was continued until diagnosis of diabetes, death or until 31 December 2005. Mean follow-up was 27.7 +/- 11.3 months. RESULTS: Among subjects enrolled, 13.7 and 25.2% were affected by MS using NCEP and IDF criteria respectively. During follow-up, 38 new cases of diabetes were diagnosed, with a yearly incidence rate of 0.5%. The relative risk for diabetes in subjects with MS was 10.10 [5.13; 20.00] and 7.87 [3.70; 16.7] using NCEP and IDF definitions respectively. After adjustment for age, sex, fasting glucose and waist circumference, NCEP-defined MS, but not IDF-, was significantly associated with incident diabetes (hazard ratio, 95% CI: 2.41 [1.01; 5.95] and 2.05 [0.80; 5.29] respectively). CONCLUSIONS: Although the reasons for the proposed changes in diagnostic criteria for MS are easily understandable, the newer IDF definition, while increasing estimates of prevalence of the syndrome, reduces the effectiveness of MS in identifying subjects at risk for diabetes. Further research is needed before the previous NCEP criteria are abandoned. PMID- 17419723 TI - NEMO oligomerization in the dynamic assembly of the IkappaB kinase core complex. AB - NF-kappaB essential modulator (NEMO) plays an essential role in the nuclear factor kappaB (NF-kappaB) pathway as a modulator of the two other subunits of the IkappaB kinase (IKK) complex, i.e. the protein kinases, IKKalpha and IKKbeta. Previous reports all envision the IKK complex to be a static entity. Using glycerol-gradient ultracentrifugation, we observed stimulus-dependent dynamic IKK complex assembly. In wild-type fibroblasts, the kinases and a portion of cellular NEMO associate in a 350-kDa high-molecular-mass complex. In response to constitutive NF-kappaB stimulation by Tax, we observed NEMO recruitment and oligomerization to a shifted high-molecular-mass complex of 440 kDa which displayed increased IKK activity. This stimulus-dependent oligomerization of NEMO was also observed using fluorescence resonance energy transfer after a transient pulse with interleukin-1beta. In addition, fully activated, dimeric kinases not bound to NEMO were detected in these Tax-activated fibroblasts. By glycerol gradient ultracentrifugation, we also showed that: (a) in fibroblasts deficient in IKKalpha and IKKbeta, NEMO predominantly exists as a monomer; (b) in NEMO deficient fibroblasts, IKKbeta dimers are present that are less stable than IKKalpha dimers. Intriguingly, in resting Rat-1 fibroblasts, 160-kDa IKKalpha NEMO and IKKbeta-NEMO heterocomplexes were observed as well as a significant proportion of NEMO monomer. These results suggest that most NEMO molecules do not form a tripartite IKK complex with an IKKalpha-IKKbeta heterodimer as previously reported in the literature but, instead, NEMO is able to form a complex with the monomeric forms of IKKalpha and IKKbeta. PMID- 17419724 TI - It's cheap to be colorful. Anthozoans show a slow turnover of GFP-like proteins. AB - Pigments homologous to the green fluorescent protein (GFP) contribute up to approximately 14% of the soluble protein content of many anthozoans. Maintenance of such high tissue levels poses a severe energetic penalty to the animals if protein turnover is fast. To address this as yet unexplored issue, we established that the irreversible green-to-red conversion of the GFP-like pigments from the reef corals Montastrea cavernosa (mcavRFP) and Lobophyllia hemprichii (EosFP) is driven by violet-blue radiation in vivo and in situ. In the absence of photoconverting light, we subsequently tracked degradation of the red-converted forms of the two proteins in coral tissue using in vivo spectroscopy and immunochemical detection of the post-translational peptide backbone modification. The pigments displayed surprisingly slow decay rates, characterized by half-lives of approximately 20 days. The slow turnover of GFP-like proteins implies that the associated energetic costs for being colorful are comparatively low. Moreover, high in vivo stability makes GFP-like proteins suitable for functions requiring high pigment concentrations, such as photoprotection. PMID- 17419725 TI - Biochemical and structural characterization of mammalian-like purine nucleoside phosphorylase from the Archaeon Pyrococcus furiosus. AB - We report here the characterization of the first mammalian-like purine nucleoside phosphorylase from the hyperthermophilic archaeon Pyrococcus furiosus (PfPNP). The gene PF0853 encoding PfPNP was cloned and expressed in Escherichia coli and the recombinant protein was purified to homogeneity. PfPNP is a homohexamer of 180 kDa which shows a much higher similarity with 5'-deoxy-5'-methylthioadenosine phosphorylase (MTAP) than with purine nucleoside phosphorylase (PNP) family members. Like human PNP, PfPNP shows an absolute specificity for inosine and guanosine. PfPNP shares 50% identity with MTAP from P. furiosus (PfMTAP). The alignment of the protein sequences of PfPNP and PfMTAP indicates that only four residue changes are able to switch the specificity of PfPNP from a 6-oxo to a 6 amino purine nucleoside phosphorylase still maintaining the same overall active site organization. PfPNP is highly thermophilic with an optimum temperature of 120 degrees C and is characterized by extreme thermodynamic stability (T(m), 110 degrees C that increases to 120 degrees C in the presence of 100 mm phosphate), kinetic stability (100% residual activity after 4 h incubation at 100 degrees C), and remarkable SDS-resistance. Limited proteolysis indicated that the only proteolytic cleavage site is localized in the C-terminal region and that the C terminal peptide is not necessary for the integrity of the active site. By integrating biochemical methodologies with mass spectrometry we assigned three pairs of intrasubunit disulfide bridges that play a role in the stability of the enzyme against thermal inactivation. The characterization of the thermal properties of the C254S/C256S mutant suggests that the CXC motif in the C terminal region may also account for the extreme enzyme thermostability. PMID- 17419726 TI - Caenorhabditis elegans expresses a functional ArsA. AB - Because arsenic is the most prevalent environmental toxin, it is imperative that we understand the mechanisms of metalloid detoxification. In prokaryotes, arsenic detoxification is accomplished by chromosomal and plasmid-borne operon-encoded efflux systems. Bacterial ArsA ATPase is the catalytic component of an oxyanion pump that is responsible for resistance to arsenite (As(III)) and antimonite (Sb(III)). Here, we describe the identification of a Caenorhabditis elegans homolog (asna-1) that encodes the ATPase component of the Escherichia coli As(III) and Sb(III) transporter. We evaluated the responses of wild-type and asna 1-mutant nematodes to various metal ions and found that asna-1-mutant nematodes are more sensitive to As(III) and Sb(III) toxicity than are wild-type animals. These results provide evidence that ASNA-1 is required for C. elegans' defense against As(III) and Sb(III) toxicity. A purified maltose-binding protein (MBP) ASNA-1 fusion protein was biochemically characterized, and its properties compared with those of ArsAs. The ATPase activity of the ASNA-1 protein was dependent on the presence of As(III) or Sb(III). As(III) stimulated ATPase activity by 2 +/- 0.2-fold, whereas Sb(III) stimulated it by 4.6 +/- 0.15-fold. The results indicate that As(III)- and Sb(III)-stimulated ArsA ATPase activities are not restricted to bacteria, but extend to animals, by demonstrating that the asna-1 gene from the nematode, C. elegans, encodes a functional ArsA ATPase whose activity is stimulated by As(III) and Sb(III) and which is critical for As(III) and Sb(III) tolerance in the intact organism. PMID- 17419728 TI - Modeling hydration mechanisms of enzymes in nonpolar and polar organic solvents. AB - A comprehensive study of the hydration mechanism of an enzyme in nonaqueous media was done using molecular dynamics simulations in five organic solvents with different polarities, namely, hexane, 3-pentanone, diisopropyl ether, ethanol, and acetonitrile. In these solvents, the serine protease cutinase from Fusarium solani pisi was increasingly hydrated with 12 different hydration levels ranging from 5% to 100% (w/w) (weight of water/weight of protein). The ability of organic solvents to 'strip off' water from the enzyme surface was clearly dependent on the nature of the organic solvent. The rmsd of the enzyme from the crystal structure was shown to be lower at specific hydration levels, depending on the organic solvent used. It was also shown that organic solvents determine the structure and dynamics of water at the enzyme surface. Nonpolar solvents enhance the formation of large clusters of water that are tightly bound to the enzyme, whereas water in polar organic solvents is fragmented in small clusters loosely bound to the enzyme surface. Ions seem to play an important role in the stabilization of exposed charged residues, mainly at low hydration levels. A common feature is found for the preferential localization of water molecules at particular regions of the enzyme surface in all organic solvents: water seems to be localized at equivalent regions of the enzyme surface independently of the organic solvent employed. PMID- 17419729 TI - Crystal structure of the tetrameric inositol 1-phosphate phosphatase (TM1415) from the hyperthermophile, Thermotoga maritima. AB - The structure of the first tetrameric inositol monophosphatase (IMPase) has been solved. This enzyme, from the eubacterium Thermotoga maritima, similarly to its archaeal homologs exhibits dual specificity with both IMPase and fructose-1,6 bisphosphatase activities. The tetrameric structure of this unregulated enzyme is similar, in its quaternary assembly, to the allosterically regulated tetramer of fructose-1,6-bisphosphatase. The individual dimers are similar to the human IMPase. Additionally, the structures of two crystal forms of IMPase show significant differences. In the first crystal form, the tetrameric structure is symmetrical, with the active site loop in each subunit folded into a beta-hairpin conformation. The second form is asymmetrical and shows an unusual structural change. Two of the subunits have the active site loop folded into a beta-hairpin structure, whereas in the remaining two subunits the same loop adopts an alpha helical conformation. PMID- 17419730 TI - Discovery of a eugenol oxidase from Rhodococcus sp. strain RHA1. AB - A gene encoding a eugenol oxidase was identified in the genome from Rhodococcus sp. strain RHA1. The bacterial FAD-containing oxidase shares 45% amino acid sequence identity with vanillyl alcohol oxidase from the fungus Penicillium simplicissimum. Eugenol oxidase could be expressed at high levels in Escherichia coli, which allowed purification of 160 mg of eugenol oxidase from 1 L of culture. Gel permeation experiments and macromolecular MS revealed that the enzyme forms homodimers. Eugenol oxidase is partly expressed in the apo form, but can be fully flavinylated by the addition of FAD. Cofactor incorporation involves the formation of a covalent protein-FAD linkage, which is formed autocatalytically. Modeling using the vanillyl alcohol oxidase structure indicates that the FAD cofactor is tethered to His390 in eugenol oxidase. The model also provides a structural explanation for the observation that eugenol oxidase is dimeric whereas vanillyl alcohol oxidase is octameric. The bacterial oxidase efficiently oxidizes eugenol into coniferyl alcohol (KM=1.0 microM, kcat=3.1 s-1). Vanillyl alcohol and 5-indanol are also readily accepted as substrates, whereas other phenolic compounds (vanillylamine, 4-ethylguaiacol) are converted with relatively poor catalytic efficiencies. The catalytic efficiencies with the identified substrates are strikingly different when compared with vanillyl alcohol oxidase. The ability to efficiently convert eugenol may facilitate biotechnological valorization of this natural aromatic compound. PMID- 17419731 TI - Plethodontid modulating factor, a hypervariable salamander courtship pheromone in the three-finger protein superfamily. AB - The soluble members of the three-finger protein superfamily all share a relatively simple 'three-finger' structure, yet perform radically different functions. Plethodontid modulating factor (PMF), a pheromone protein produced by the lungless salamander, Plethodon shermani, is a new and unusual member of this group. It affects female receptivity when delivered to the female's nares during courtship. As with other plethodontid pheromone genes, PMF is hyperexpressed in a specialized male mental (chin) gland. Unlike other plethodontid pheromone genes, however, PMF is also expressed at low levels in the skin, liver, intestine and kidneys of both sexes. The PMF sequences obtained from all tissue types were highly variable, with 103 unique haplotypes identified which averaged 35% sequence dissimilarity (range 1-60%) at the protein level. Despite this variation, however, all PMF sequences contained a conserved approximately 20 amino-acid secretion signal sequence and a pattern of eight cysteines that is also found in cytotoxins and short neurotoxins from snake venoms, as well as xenoxins from Xenopus. Although they share a common cysteine pattern, PMF isoforms differ from other three-finger proteins in: (a) amino-acid composition outside of the conserved motif; (b) length of the three distinguishing 'fingers'; (c) net charge at neutral pH. Whereas most three-finger proteins have a net positive charge at pH 7.0, PMF has a high net negative charge at neutral pH (pI range of most PMFs 3.5-4.0). Sequence comparisons suggest that PMF belongs to a distinct multigene subfamily within the three-finger protein superfamily. PMID- 17419732 TI - A novel tachykinin-related peptide receptor of Octopus vulgaris--evolutionary aspects of invertebrate tachykinin and tachykinin-related peptide. AB - The tachykinin (TK) and tachykinin-related peptide (TKRP) family represent one of the largest peptide families in the animal kingdom and exert their actions via a subfamily of structurally related G-protein-coupled receptors. In this study, we have identified a novel TKRP receptor from the Octopus heart, oct-TKRPR. oct TKRPR includes domains and motifs typical of G-protein-coupled receptors. Xenopus oocytes that expressed oct-TKRPR, like TK and TKRP receptors, elicited an induction of membrane chloride currents coupled to the inositol phosphate/calcium pathway in response to Octopus TKRPs (oct-TKRP I-VII) with moderate ligand selectivity. Substance P and Octopus salivary gland-specific TK, oct-TK-I, completely failed to activate oct-TKRPR, whereas a Substance P analog containing a C-terminal Arg-NH2 exhibited equipotent activation of oct-TKRPs. These functional analyses prove that oct-TKRPs, but not oct-TK-I, serve as endogenous functional ligands through oct-TKRPR, although both of the family peptides were identified in a single species, and the importance of C-terminal Arg-NH2 in the specific recognition of TKRPs by TKRPR is conserved through evolutionary lineages of Octopus. Southern blotting of RT-PCR products revealed that the oct-TKRPR mRNA was widely distributed in the central and peripheral nervous systems plus several peripheral tissues. These results suggest multiple physiologic functions of oct TKRPs as neuropeptides both in the Octopus central nervous system and in peripheral tissues. This is the first report on functional discrimination between invertebrate TKRPs and salivary gland-specific TKs. PMID- 17419733 TI - Autonomous folding of interdomain regions of a modular polyketide synthase. AB - Domains within the multienzyme polyketide synthases are linked by noncatalytic sequences of variable length and unknown function. Recently, the crystal structure was reported of a portion of the linker between the acyltransferase (AT) and ketoreductase (KR) domains from module 1 of the erythromycin synthase (6 deoxyerythronolide B synthase), as a pseudodimer with the adjacent ketoreductase (KR). On the basis of this structure, the homologous linker region between the dehydratase (DH) and enoyl reductase (ER) domains in fully reducing modules has been proposed to occupy a position on the periphery of the polyketide synthases complex, as in porcine fatty acid synthase. We report here the expression and characterization of the same region of the 6-deoxyerythronolide B synthase module 1 AT-KR linker, without the adjacent KR domain (termed DeltaN AT1-KR1), as well as the corresponding section of the DH-ER linker. The linkers fold autonomously and are well structured. However, analytical gel filtration and ultracentrifugation analysis independently show that DeltaN AT1-KR1 is homodimeric in solution; site-directed mutagenesis further demonstrates that linker self-association is compatible with the formation of a linker-KR pseudodimer. Our data also strongly indicate that the DH-ER linker associates with the upstream DH domain. Both of these findings are incompatible with the proposed model for polyketide synthase architecture, suggesting that it is premature to allocate the linker regions to a position in the multienzymes based on the solved structure of animal fatty acid synthase. PMID- 17419734 TI - The activity of adenylyltransferase in Rhodospirillum rubrum is only affected by alpha-ketoglutarate and unmodified PII proteins, but not by glutamine, in vitro. AB - Ammonium assimilation is tightly regulated in nitrogen-fixing bacteria; the target of regulation is primarily the activity of the key enzyme glutamine synthetase that is regulated by reversible covalent modification by AMP groups in reactions catalysed by the bifunctional adenylyltransferase (ATase). The properties and regulation of ATase from Escherichia coli have been studied in great detail. We have investigated the regulation of ATase from Rhodospirillum rubrum, a photosynthetic nitrogen-fixing bacterium. In this diazotroph, nitrogenase is regulated at the metabolic level in addition to the transcriptional regulation operating in all diazotrophic bacteria, which makes understanding the regulatory features of nitrogen assimilation even more interesting. We show that in R. rubrum, in contrast to the E. coli system, ATase is primarily regulated by alpha-ketoglutarate and that glutamine has no effect on neither the adenylylation nor the deadenylylation of glutamine synthetase. Furthermore, the role of the regulatory P(II) proteins is only to stimulate the adenylylation reaction, as there is no effect on the reverse reaction. We propose that in R. rubrum and possibly other diazotrophs alpha-ketoglutarate plays the central role in the regulation of ATase and thus glutamine synthetase activity. PMID- 17419735 TI - Two conserved regions within the tissue-type plasminogen activator gene promoter mediate regulation by brain-derived neurotrophic factor. AB - Tissue-type plasminogen activator (t-PA) has recently been identified as a modulator of neuronal plasticity and can initiate conversion of the pro-form of brain-derived neurotrophic factor (BDNF) into its mature form. BDNF also increases t-PA gene expression implicating t-PA as a downstream effector of BDNF function. Here we demonstrate that BDNF-mediated induction of t-PA mRNA requires an increase in t-PA gene transcription. Reporter constructs harboring 9.5 kb of the human t-PA promoter conferred BDNF-responsiveness in transfected mouse primary cortical neurons. This regulation was recapitulated in HEK 293 cells coexpressing the TrkB neurotrophin receptor. t-PA promoter-deletion analysis revealed the presence of two BDNF-responsive domains, one located between -3.07 and -2.5 kb and the other within the proximal promoter. The upstream region was shown to confer BDNF responsiveness in a TrkB-dependent manner when attached to a heterologous promoter. We also identify homologous regions within the murine and bovine t-PA gene promoters and demonstrate that the equivalent upstream murine sequence functions as a BDNF-responsive enhancer when inserted 5' of the human proximal t-PA promoter. Hence, BDNF-mediated induction of t-PA transcription relies on conserved modular promoter elements including a novel upstream BDNF responsive domain and the proximal t-PA gene promoter. PMID- 17419736 TI - Nucleosome positioning in relation to nucleosome spacing and DNA sequence specific binding of a protein. AB - Nucleosome positioning is an important mechanism for the regulation of eukaryotic gene expression. Folding of the chromatin fiber can influence nucleosome positioning, whereas similar electrostatic mechanisms govern the nucleosome repeat length and chromatin fiber folding in vitro. The position of the nucleosomes is directed either by the DNA sequence or by the boundaries created due to the binding of certain trans-acting factors to their target sites in the DNA. Increasing ionic strength results in an increase in nucleosome spacing on the chromatin assembled by the S-190 extract of Drosophila embryos. In this study, a mutant lac repressor protein R3 was used to find the mechanisms of nucleosome positioning on a plasmid with three R3-binding sites. With increasing ionic strength in the presence of R3, the number of positioned nucleosomes in the chromatin decreased, whereas the internucleosomal spacings of the positioned nucleosomes in a single register did not change. The number of the positioned nucleosomes in the chromatin assembled in vitro over different plasmid DNAs with 1-3 lac operators changed with the relative position and number of the R3-binding sites. We found that in the presence of R3, nucleosomes were positioned in the salt gradient method of the chromatin assembly, even in the absence of a nucleosome-positioning sequence. Our results show that nucleosome-positioning mechanisms are dominant, as the nucleosomes can be positioned even in the absence of regular spacing mechanisms. The protein-generated boundaries are more effective when more than one binding site is present with a minimum distance of approximately 165 bp, greater than the nucleosome core DNA length, between them. PMID- 17419737 TI - Seleno-independent glutathione peroxidases. More than simple antioxidant scavengers. AB - Glutathione peroxidases (GPXs, EC 1.11.1.9) were first discovered in mammals as key enzymes involved in scavenging of activated oxygen species (AOS). Their efficient antioxidant activity depends on the presence of the rare amino-acid residue selenocysteine (SeCys) at the catalytic site. Nonselenium GPX-like proteins (NS-GPXs) with a Cys residue instead of SeCys have also been found in most organisms. As SeCys is important for GPX activity, the function of the NS GPX can be questioned. Here, we highlight the evolutionary link between NS-GPX and seleno-GPX, particularly the evolution of the SeCys incorporation system. We then discuss what is known about the enzymatic activity and physiological functions of NS-GPX. Biochemical studies have shown that NS-GPXs are not true GPXs; notably they reduce AOS using reducing substrates other than glutathione, such as thioredoxin. We provide evidence that, in addition to their inefficient scavenging action, NS-GPXs act as AOS sensors in various signal-transduction pathways. PMID- 17419738 TI - Loss of ATP hydrolysis activity by CcmAB results in loss of c-type cytochrome synthesis and incomplete processing of CcmE. AB - The proteins CcmA and CcmB have long been known to be essential for cytochrome c maturation in Escherichia coli. We have purified a complex of these proteins, and found it to have ATP hydrolysis activity. CcmA, which has the features of a soluble ATP hydrolysis subunit, is found in a membrane-bound complex only when CcmB is present in the membrane. Mutation of the Walker A motif in CcmA(K40D) results in loss of the in vitro ATPase activity and in loss of cytochrome c biogenesis in vivo. The same mutation does not prevent covalent attachment of heme to the heme chaperone CcmE, but holo-CcmE is, for some unidentified reason, incompetent for heme transfer to an apocytochrome c or for release into the periplasm as a soluble variant. Addition of exogenous heme to heme-permeable E. coli with a ccmA deletion did not restore cytochrome c production. Our results suggest a role for CcmAB in the handling of heme by CcmE, which is chemically complex and involves an unusual histidine-heme covalent bond. PMID- 17419739 TI - Lysophosphatidic acid protection against apoptosis in the human pre-B-cell line Nalm-6. AB - Lysophosphatidic acid (LPA) promotes survival, growth, differentiation, and motility in a variety of cell types, and has been reported to act as a cell survival and growth factor in B lymphocytes. Autotaxin (ATX), through its lysophospholipase D activity, generates LPA from lysophosphatidylcholine (LPC). In this study, we investigated the effects of LPA and also the expression of ATX and LPA receptor, in the human pre-B-cell line Nalm-6. It was found that LPA protects Nalm-6 cells against both spontaneous and staurosporine-induced apoptosis. Furthermore, ATX expression on the cell surface and ATX activity in the cell lysate were detected. No accumulation of LPA in the culture medium was, however, detected when the Nalm-6 cells were cultured with LPC. The pre-B cells were found to express the mRNA transcript for lipid phosphate phosphatase-1 and LPA degradation was inhibited in the presence of the phosphatase inhibitor vanadate, it was surmised that LPA production in the culture medium may have been masked by LPA degradation by this ecto-phosphatase. Abundant expression of LPA receptors, especially, LPA(4), was detected by a real-time polymerase chain reaction technique. Our results suggest an important and autocrine role of LPA in the survival of this well-established model cell line, although the direct involvement of ATX in the production of LPA in these cells was not confirmed. PMID- 17419740 TI - Health-related quality of life measured by the Short Form 36 in immune thrombocytopenic purpura: a cross-sectional survey in China. AB - OBJECTIVES: The aim of this study is to assess the quality of life (QoL) of Chinese adults with idiopathic thrombocytopenic purpura (ITP). METHODS: The Chinese (mainland) version of Medical Outcome Study SF-36 form (SF-36) Health Survey was used to measure health-related QoL of 236 adults with ITP in a cross sectional study. RESULTS: Comparison of SF-36 subscores of patients with ITP with healthy individuals revealed the reduction of QoL in all of the eight SF-36 dimensions. The difference on statistical significance presented in six of eight dimensions of SF-36 including physical functioning (PF), role limitations due to physical problems, body pain, general health perception (GH), social functioning (SF), and role limitations due to emotional problems (RE) between the patients with ITP and the normal population (P < 0.01). The acute ITP group showed better scores in three dimensions including GH, energy/vitality, and RE than chronic ITP (P < 0.01). Meanwhile through classification with platelet count, three subgroups of patients also experienced significant differences in PF, GH, and SF from the eight dimensions. Age was a significant negative predictor of all eight dimensions other than the SF while current platelet count was a significant negative predictor of GH. Moreover, the treatment cost and family income also influenced the QoL scores. The subjective feeling of fear about bleeding had a detrimental impact on QoL. CONCLUSIONS: QoL was impaired in patients with ITP, especially in the acute patients. The platelet count and the feeling of fear about bleeding had a detrimental impact on QoL. PMID- 17419741 TI - Limited usefulness of CA125 measurement in the management of Hodgkin's and non Hodgkin's lymphoma. AB - BACKGROUND: Several papers have reported an association of high CA125 serum levels with advanced non-Hodgkin's lymphoma (NHL) as well as a relationship between high CA125 values and poor outcome. PATIENTS AND METHODS: Ninety-nine patients with NHL or Hodgkin's disease (HD) underwent serum CA125 assessment at diagnosis. Gender, age, presence of B symptoms, performance status (PS), histology, sites of tumor involvement, presence of effusion, clinical stage, age adjusted International Prognostic Index, C-reactive protein (CRP), Hb, lactate deshydrogenase (LDH) and beta2-microglobulin were evaluated for their association with serum CA125 levels. The impact of CA125 levels and other features on overall (OS) and progression-free (PFS) survival was also assessed. RESULTS: CA125 serum levels were elevated in 34% of the patients, including 19% of patients with aggressive NHL, 45% of patients with indolent NHL, and 29% of patients with HD. Univariate analyses showed that CA125 levels correlated with poor PS, the presence of B symptoms, advanced clinical stage, abdominal, bone marrow or mediastinal involvement, presence of effusions, high aaIPI, low Hb levels and high CRP, LDH or beta2-microglobulin levels. In multivariate analysis, bone marrow involvement, the presence of effusions, and high aaIPI were all associated with high CA125 serum levels. In univariate analyses, OS and PFS were affected by age (PFS only), poor PS, B symptoms, advanced clinical stage, bone marrow or abdominal involvement (PFS only), high aaIPI, low Hb, high CRP or beta2 microglobulin levels. OS and PFS were not different in patients with normal or elevated CA125 levels. Multivariate analyses showed significantly inferior OS and PFS in patients with high beta2-microglobulin but no influence of CA125. CONCLUSION: While CA125 serum level correlates significantly with a number of features associated with more aggressive disease, it does not enhance the performance of standard prognostic markers in the management of patients with NHL or HD. PMID- 17419742 TI - Predictive value of discrimination indices in differential diagnosis of iron deficiency anemia and beta-thalassemia trait. AB - OBJECTIVES: Iron deficiency anemia (IDA) and beta-thalassemia trait (B-TT) are the most common causes of hypochromic microcytic anemias. Many indices have been defined to quickly discriminate these similar entities via parameters obtained from automated blood count analyzers. The purpose of the study was to evaluate the predictive value of these indices in differential diagnosis of IDA and B-TT in adult cases. METHODS: This study consists of 45 IDA cases, 36 women and nine men, whose mean age is 33.87 +/- 11.59 (mean +/- SD) (range 17-57 yr) and 66 B-TT cases, 41 women and 25 men, whose mean age is 33.26 +/- 13.36 (mean +/- SD) (range 14-74 yr). IDA cases with Hb value <8.7 g/dL have been excluded because these cases are not confused with B-TT cases in practice. Patient groups have been evaluated according to red blood cell (RBC), red blood cell distribution width (RDW), Mentzer index, Shine and Lal indices, England and Fraser indices, Srivastava index, Green and King indices, RDW index and Ricerca index. Sensitivity, specificity, positive and negative predictive values and Youden's index have been calculated. RESULTS: Finally, none of these different formulations are superior to RBC value obtained from automated analyzers in adult cases with IDA and B-TT. CONCLUSIONS: Total body iron status and hemoglobin A(2) level should be obtained for accurate differential diagnosis of IDA and B-TT until more efficient tools develop. PMID- 17419743 TI - Bone changes in myelofibrosis with myeloid metaplasia: a histomorphometric and microcomputed tomographic study. AB - Myelofibrosis with myeloid metaplasia (MMM) is a clonal disorder of the haematopoietic stem cell which can be associated with marrow fibrosis and/or osteosclerosis. Because bone progenitors and mature bone cells are influenced by the marrow microenvironment, cellular and tissular changes were assessed by histomorphometry in MMM. Thirteen patients, with a clinical proven MMM, had a bone biopsy of the iliac crest with double tetracycline labelling and osteoclast count. Histomorphometry was done at the 2D level (bone volume, osteoid parameters, bone histodynamic parameters and osteoclast count) and 3D level by microcomputed tomography. All patients had clusters of abnormal megakaryocytes in bone marrow. Newly apposed bone packets were observed in 12 patients and corresponded to an increased thickness of some bone units with new lamellae or focal areas of woven bone anchored on the pre-existing trabeculae. Osteoid parameters were unchanged, only bone formation rate appeared considerably increased in seven patients. There was a net tendency for decrease in osteoclast number and conversion of trabecular pillars into plates. An uncoupling of bone remodelling was evidenced with an increased life-span of osteoblasts associated with a normal/reduced osteoclast activity. A very complex network of factors is candidate to explain bone changes observed in MMM. PMID- 17419744 TI - A cost-effectiveness analysis of caspofungin vs. liposomal amphotericin B for treatment of suspected fungal infections in the UK. AB - OBJECTIVE: To evaluate the cost-effectiveness of caspofungin vs. liposomal amphotericin B in the treatment of suspected fungal infections in the UK. METHODS: The cost-effectiveness of caspofungin vs. liposomal amphotericin B was evaluated using a decision-tree model. The decision tree was populated using both data and clinical definitions from published clinical studies. Model outcomes included success in terms of resolution of fever, baseline infection, absence of breakthrough infection, survival and quality adjusted life years (QALYs) saved. Discontinuation due to nephrotoxicity or other adverse events were included in the model. Efficacy and safety data were based on additional analyses of a randomised, double blind, multinational trial of caspofungin compared with liposomal amphotericin B. Information on life expectancy, quality of life, medical resource consumption and costs were obtained from peer-reviewed published data. RESULTS: The caspofungin mean total treatment cost was 9762 pounds (95% uncertainty interval 6955-12,577), which was 2033 pounds (-2489; 6779) less than liposomal amphotericin B. Treatment with caspofungin resulted in 0.40 (-0.12; 0.94) additional QALYs saved in comparison with liposomal amphotericin B. Probabilistic sensitivity analysis found a 95% probability of the incremental cost per QALY saved being within the generally accepted threshold for cost effectiveness (30,000 pounds). Additional analyses with varying dose of caspofungin and liposomal amphotericin B confirmed these findings. CONCLUSION: Given the underlying assumptions, caspofungin is cost-effective compared with liposomal amphotericin B in the treatment of suspected fungal infections in the UK. PMID- 17419745 TI - CHOP chemotherapy followed by Rituximab consolidation as first line treatment in patients with follicular lymphoma. Long-term follow-up of a phase 2 trial. PMID- 17419746 TI - Bullous pyoderma gangrenosum in acute myeloid leukaemia. PMID- 17419747 TI - Omission of evidence from review on prophylaxis of fungal infections. PMID- 17419749 TI - Rapid elimination of circulating leukemia cells with a small dose of alemtuzumab in refractory aggressive NK-cell leukemia. PMID- 17419750 TI - The prognostic significance of cytogenetic aberrations in childhood acute myeloid leukaemia. A study of the Swiss Paediatric Oncology Group (SPOG). AB - In childhood-onset acute myeloid leukaemia (AML) the clinical value of karyotypic aberrations is now acknowledged, although there is still debate concerning the prognostic significance of some events. To add to this knowledge, cytogenetic analysis was performed on a consecutive series of 84 childhood AML patients diagnosed in Switzerland. A result was obtained for all patients, with 69 (82%) showing a clonal karyotypic aberration. In the remaining 15 (18%), no karyotypic aberration was seen by either conventional or fluorescence in situ hybridisation analyses. The most frequent aberrations observed were t(11q23) (19% of all patients), t(8;21) (12%) and +8 (11%). Except for cytogenetics, no clinical parameter was shown to be significantly associated with outcome. The analysis of individual cytogenetic subgroups demonstrated that aberrations involving chromosome 16q were the strongest predictor of a good prognosis, while +8 and complex karyotypes represented the strongest predictors of a poor prognosis. It was also noteworthy that patients with the rare aberrations of del(11q) (n = 4) and t(16;21)(p11;q22) (n = 3) had a poor outcome. The results support the importance of cytogenetic analysis in childhood AML, but show that further work is required in the classification of the poor prognosis aberrations. PMID- 17419751 TI - Heterogeneous population of dopaminergic neurons derived from mouse embryonic stem cells: preliminary phenotyping based on receptor expression and function. AB - The possibility exists that directed differentiation of mouse embryonic stem (mES) cells is capable of yielding enriched populations of dopaminergic neurons, but at present there is little understanding of the pharmacological properties of these cells; or whether such cells represent a pharmacologically, phenotypically similar population. In this study we used a simple culture protocol to generate dopaminergic neurons and offer a preliminary pharmacological investigation of these cells using Ca2+ imaging and [3H]-dopamine release studies. In fluo-4 AM loaded cells, 13-17 days postplating, and after the addition of tetrodotoxin some of the population of mouse embryonic stem cell-derived neurons responded to adenosine triphosphate (ATP), noradrenaline (NA), acetylcholine (ACh) and L glutamate (L-glut) with elevations of Ca2+ influx. Within the microtubule associated protein and tyrosine hydroxylase (TH)-positive cell population adenosine triphosphate, noradrenaline, acetylcholine and L-glutamate elicited positive elevations of Ca2+ in 74, 66, 58 and 67% of the population; cells could be further subdivided into three major pharmacologically distinct populations based on the combinations of agonist they responded to. Acetylcholine (30 microM) and noradrenaline (30 microM) were the only agonists to elicit significant tritium overflow from [3H]-dopamine loaded cells. The acetylcholine effect was blocked by atropine (1 microM) and tetrodotoxin (1 microM) and elevated by haloperidol (100 nM). The noradrenaline effects were reduced by cocaine (10 microM), but not by tetrodotoxin (100 nM). These data indicate that the dopaminergic neurons derived from mouse embryonic stem cells represent a heterogeneous population possessing combinations of purinergic, adrenergic, cholinergic and glutamatergic receptors located on the cell soma. PMID- 17419752 TI - Androgen receptor is essential for sexual differentiation of responses to olfactory cues in mice. AB - During sexual differentiation males and females are exposed to different levels of testosterone, which promotes sex differences in the adult brain and in behavior. Testosterone can act after aromatization or reduction via a number of steroid hormone receptors. Here we provide new evidence that the androgen receptor (AR) is essential for sexual differentiation in mice. We used mice carrying the testicular feminization (Tfm) mutation of the AR. Adult Tfm males, wild-type male and female littermates were gonadectomized and given subcutaneous estradiol implants. In all sexually dimorphic traits, Tfm males had responses equivalent to females and different from males. In simultaneous choice tests, males spent significantly more time investigating female-soiled bedding, whereas females and Tfm males preferred to investigate male-soiled bedding. Tfm males and females did not have a partner preference in tests with awake stimulus animals, whereas males showed a preference for females over males. Exposure to male soiled, but not clean, bedding produced a significant increase in c-Fos immunoreactive cells in the medial preoptic area and bed nucleus of the stria terminalis in Tfm males and females, no increase was noted in males. Masculine sexual behavior (mounting and thrusting) was not sexually dimorphic, and all groups displayed these behaviors. Our results support data collected in humans suggesting a role for the androgen receptor in sexual differentiation of social preferences and neural responses to pheromones. PMID- 17419753 TI - Accurate visual guidance despite severe neglect. AB - Unilateral inactivation of the superior colliculus causes profound neglect. In cats, this neglect has been studied previously using tasks that require gaze orientation to, or detection of, a stimulus appearing somewhere in the visual field of an attentive animal. We investigated how neglect affects a completely different kind of task, visually guided foot placement while walking across a cluttered surface. We made muscimol injections into one superior colliculus, and performed perimetry to gauge the extent of the cat's neglect. Cats then walked repeatedly through a cluttered test alley. Most of the time, their gaze was deviated towards the side of the injection, so that they saw the alley floor ahead of them in their neglected hemifield. Surprisingly, they accurately avoided stepping on the densely scattered objects, just as they normally do. We surmise that cats process 'neglected' visual stimuli to guide foot placement even when they are unable to consciously perceive these stimuli. PMID- 17419754 TI - Segregated pathways to the vomeronasal amygdala: differential projections from the anterior and posterior divisions of the accessory olfactory bulb. AB - Apically and basally located receptor neurons in the vomeronasal sensory epithelium express G(i2 alpha)- and G(o alpha)-proteins, V1R and V2R vomeronasal receptors, project to the anterior and posterior accessory olfactory bulb and respond to different stimuli, respectively. The extent to which secondary projections from the two portions of the accessory olfactory bulb are convergent in the vomeronasal amygdala is controversial. This issue is addressed by using anterograde and retrograde tract-tracing methods in rats including electron microscopy. Injections of dextran-amines, Fluoro Gold, cholera toxin-B subunit and Fast Blue were delivered to the anterior and posterior accessory olfactory bulb, bed nucleus of the stria terminalis, dorsal anterior amygdala and bed nucleus of the accessory olfactory tract/anteroventral medial amygdaloid nucleus. We have demonstrated that, apart from common vomeronasal-recipient areas, only the anterior accessory olfactory bulb projects to the bed nucleus of the stria terminalis, medial division, posteromedial part, and only the posterior accessory olfactory bulb projects to the dorsal anterior amygdala and deep cell layers of the bed nucleus of the accessory olfactory tract and the anteroventral medial amygdaloid nucleus. These results provide evidence that, excluding areas of convergence, the V1R and V2R vomeronasal pathways project to specific areas of the amygdala. These two vomeronasal subsystems are therefore anatomically and functionally separated in the telencephalon. PMID- 17419755 TI - Bidirectional regulation of mu-opioid and CB1-cannabinoid receptor in rats self administering heroin or WIN 55,212-2. AB - This study examines the effect of intravenous self-administration (SA) of either heroin or the cannabinoid receptor agonist WIN 55,212-2 on levels and functionality of mu-opioid (MOR) and CB1-cannabinoid receptors (CB1R) in reward related brain areas, such as the prefrontal cortex (PFC), nucleus accumbens (NAc), caudate putamen (CP), hippocampus (Hippo), amygdala (Amy), hypothalamus (Hypo) and ventral tegmental area (VTA). [3H]DAMGO and [3H]CP-55,940 autoradiography and agonist-stimulated [35S]GTPgammaS binding were performed on brain sections of rats firmly self-administering heroin or WIN 55,212-2. Animals failing to acquire heroin or cannabinoid SA behaviour as well as drug-naive animals never exposed to experimental apparatus or procedure (home-control group) were used as controls. With respect to control groups, which displayed very similar values, rats SA heroin showed increased MOR binding in the NAc (+174%), CP (+165%), Hippo (+121%), VTA (+175%), an enhanced CB1R density localized in the Amy (+147%) and VTA (+37%), and a widespread increased CB1 receptor functionality in the PFC (+95%), NAc (+313%), CP (+265%), Hippo (+38%), Amy (+221%). In turn, cannabinoid SA differently modulates CB1R binding in the Amy (+47%), Hypo (+94%), Hippo (-23%), VTA (-15%), and increases MOR levels (PFC: +124%; NAc: +68%; CP: +80%; Hippo: +73%; Amy: +99%) and efficiency (Hippo: +518%; Amy: +173%; Hypo: +188%). These findings suggest that voluntary chronic intake of opioids or cannabinoids induces reciprocal but differential regulation of MORs and CB1Rs density and activity in brain structures underlying drug-taking and drug-seeking behaviour, which could represent long-term neuroadaptations contributing to the development of drug addiction and dependence. PMID- 17419756 TI - Effects of the GABA(A) receptor antagonists bicuculline and gabazine on stimulus induced sharp wave-ripple complexes in adult rat hippocampus in vitro. AB - Hippocampal sharp wave-ripple-complexes (SPW-Rs) are characterized by slow field potential transients superimposed by ripples with a frequency of approximately 200 Hz. In epileptic humans and rats frequencies of such transient network oscillations can reach up to 500 Hz potentially due to loss of functional inhibition. Therefore, we investigated whether GABA(A) receptor antagonists increased ripple frequency during SPW-Rs. Within area CA3, SPW-Rs were induced by repeated stimulation of stratum radiatum in area CA1 of adult Wistar rat hippocampal slices. Intracellular recordings showed that in approximately 50% of recorded CA3 pyramidal cells SPW-Rs were accompanied by compound excitatory postsynaptic potentials (EPSPs) of approximately 10 mV superimposed by up to four action potentials (APs). The remaining cells responded with a compound inhibitory postsynaptic potential (IPSP) during SPW-Rs. The GABA(A) receptor antagonists bicuculline (BMI) or gabazine (SR-95531) led to a transition of SPW-Rs into prolonged bursts with a significant increase in amplitude and duration reminiscent of recurrent epileptiform discharges (REDs). Ripple frequencies increased from approximately 190 Hz to approximately 300 Hz. In naive slices SR 95531 and BMI also evoked REDs with similar incidence and high frequency ripple frequencies of approximately 240 Hz. Elevations in extracellular potassium concentration during REDs were approximately 20-fold higher than those observed during SPW-Rs. Intracellular recordings revealed bursts that were characterized by a large (> 25 mV) prolonged depolarization superimposed by up to 40 APs in close synchrony with extracellularly recorded ripples. Our results suggest that the generation of high frequency ripples, which are also observed in epileptic humans and rats, could indicate a loss of functional inhibition. PMID- 17419757 TI - Detection psychophysics of intracortical microstimulation in rat primary somatosensory cortex. AB - A problem of purposeful intracortical microstimulation is the long duration of neuronal integration time and the associated complex temporal interactions of effects to individual pulses in trains. Here we investigated the effects of repetitive stimuli on perception. We trained head-restraint rats to indicate the detection of cortical microstimulation in infragranular layers of barrel cortex. Three stimulus parameters: stimulus intensity, number of pulses and frequency were varied, and psychometric detection curves were assessed using the method of constant stimuli. The average psychophysical threshold of single pulses was 2.0 nC--a measure very close to what has been found earlier for the evocation of short-latency action potentials in neurons near the stimulation electrode. Detection of single-pulse stimulation always saturated at probabilities of about 0.8. In contrast, repetitive stimuli gave rise to lower thresholds (by a factor of two at 15 pulses, 320 Hz), and to saturation at probabilities close to 1. Interestingly, a large fraction of these perceptual benefits was observed already with double pulses. Moreover, the perceptual efficacy of individual pulses was higher using double pulses compared with longer sequences, i.e. double pulses were detected better than expected from the assumption of independence of single pulse effects, while trains of 15 pulses fell well short of this expectation. The present results thus point to double-pulse stimulation as an optimal choice when trading economic stimulation against optimizing of the percept. PMID- 17419758 TI - Interactions of cannabidiol with endocannabinoid signalling in hippocampal tissue. AB - The phytocannabinoid cannabidiol (CBD) possesses no psychotropic activity amid potentially beneficial therapeutic applications. We here characterized interactions between CBD (1 microM) and the endocannabinoid system in cultured rat hippocampal cells. The CBD-induced Ca2+ rise observed in neurons and glia was markedly reduced in the presence of the endogenous cannabinoid anandamide in neurons, with no alteration seen in glia. Neuronal CBD responses were even more reduced in the presence of the more abundant endocannabinoid 2-arachidonyl glycerol, this action was maintained in the presence of the CB1 receptor antagonist AM281 (100 nM). Neuronal CBD responses were also reduced by pre exposure to glutamate, expected to increase endocannabinoid levels by increasing in [Ca2+]i. Application of AM281 at 1 microM elevated CBD-induced Ca2+ responses in both cell types, further confirming our finding that endocannabinoid-mediated signalling is negatively coupled to the action of CBD. However, upregulation of endogenous levels of endocannabinoids via inhibition of endocannabinoid hydrolysis (with URB597 and MAFP) could not be achieved under resting conditions. Because delta9-tetrahydrocannabinol did not mimic the endocannabinoid actions, and pertussis toxin treatment had no effect on CBD responses, we propose that the effects of AM281 were mediated via a constitutively active signalling pathway independent of CB1 signalling. Instead, signalling via G(q/11) and phospholipase C appears to be negatively coupled to CBD-induced Ca2+ responses, as the inhibitor U73122 enhanced CBD responses. Our data highlight the interaction between exogenous and endogenous cannabinoid signalling, and provide evidence for the presence of an additional pharmacological target, sensitive to endocannabinoids and to AM281. PMID- 17419759 TI - Novel stimulus-induced calcium efflux in Drosophila mushroom bodies. AB - The mushroom body (MB) is an important part of the Drosophila brain, and is involved in many behaviors, including olfactory learning and memory and some visual cognition. However, the physiological properties of MB neurons remain elusive. Here we used a calcium-imaging technique to study calcium signals in Drosophila MB. We found that, rather than increasing calcium spread, electrical stimuli dramatically decreased calcium signals in the terminals of MB fibers. This novel calcium decrease occurred at all developmental stages from larvae to adults, but was specific for certain regions of the MB neurons. GABA receptor blockade promoted calcium propagation through the MB fibers, but did not disrupt the stimulus-induced decrease in calcium. Furthermore, this decrease in calcium was independent of extracellular calcium concentration and was not due to altered uptake by intracellular calcium stores and mitochondria. Rather, we found that inhibition of sodium-calcium exchangers significantly attenuated the stimulus induced decrease in calcium, whereas the decrease persisted when membrane calcium pumps were blocked. Our findings indicate that MB neurons exhibit a novel stimulus-induced calcium efflux, which may be importantly regulated by sodium calcium exchangers in the Drosophila MB. PMID- 17419760 TI - The pallial basal ganglia pathway modulates the behaviorally driven gene expression of the motor pathway. AB - The discrete neural network for songbird vocal communication provides an effective system to study neural mechanisms of learned motor behaviors in vertebrates. This system consists of two pathways--a vocal motor pathway used to produce learned vocalizations and a vocal pallial basal ganglia loop used to learn and modify the vocalizations. However, it is not clear how the loop exerts control over the motor pathway. To study the mechanism, we used expression of the neural activity-induced gene ZENK (or egr-1), which shows singing-regulated expression in a social context-dependent manner: high levels in both pathways when singing undirected and low levels in the lateral part of the loop and in the robust nucleus of the arcopallium (RA) of the motor pathway when singing directed to another animal. Here, we show that there are two parallel interactive parts within the pallial basal ganglia loop, lateral and medial, which modulate singing driven ZENK expression of the motor pathway nuclei RA and HVC, respectively. Within the loop, the striatal and pallial nuclei appear to have opposing roles; the striatal vocal nucleus lateral AreaX is required for high ZENK expression in its downstream nuclei, particularly during undirected singing, while the pallial vocal lateral magnocellular nucleus of the anterior nidopallium is required for lower expression, particularly during directed singing. These results suggest a dynamic molecular interaction between the basal ganglia pathway and the motor pathway during production of a learned motor behavior. PMID- 17419761 TI - Global transcriptome analysis of the heat shock response of Bifidobacterium longum. AB - Bifidobacteria are natural inhabitants of the human gastrointestinal tract and have been widely used as functional foods in different products. During industrial processing, bacterial cells undergo several stresses that can limit large-scale production and stability of the final product. To better understand the stress-response mechanisms of bifidobacteria, microarrays were used to obtain a global transcriptome profile of Bifidobacterium longum NCC2705 exposed to a heat shock treatment at 50 degrees C for 3, 7 and 12 min. Gene expression data highlighted a profound modification of gene expression, with 46% of the genes being altered. This analysis revealed a slow-down of Bi. longum general metabolic activity during stress with a simultaneous activation of the classical heat shock stimulon. Moreover, the expression of several genes with unknown function was highly induced under stress conditions. Three of these were conserved in other bacteria species where they were also previously shown to be induced by high temperature, suggesting their widespread role in the heat stress response. Finally, the implication of the trans-translation machinery in the response of Bi. longum cells to heat shock was suggested by the induction of the gene encoding the tmRNA-associated small protein B (SmpB) with concomitant high constitutive expression of the tmRNA gene. PMID- 17419762 TI - A new role of Deinococcus radiodurans RecD in antioxidant pathway. AB - In Deinococcus radiodurans, RecBCD holoenzyme is not intact because of the absence of RecB and RecC, but a RecD-like protein does indeed exist. In this work, D. radiodurans recD disruptant was constructed and its possible biological functions were investigated. The results showed that disruption of the recD gene of D. radiodurans resulted in a remarkably increased sensitivity to hydrogen peroxide but had no apparent effect on the resistance to gamma and UV radiation. Furthermore, complementation experiments showed that Escherichia coli RecD, helicase domain or N-terminal domain of D. radiodurans RecD could not individually restore the resistant phenotype to hydrogen peroxide of the recD disruptant, whereas the complete D. radiodurans RecD protein could. Further studies showed that D. radiodurans RecD took part in antioxidant process by stimulating catalase activity and reactive oxygen species scavenging activity in D. radiodurans. These results suggest that D. radiodurans RecD has a new role in the antioxidant pathway. PMID- 17419763 TI - Ultrasound screening for asymptomatic deep vein thrombosis after major orthopaedic surgery: the VENUS study. AB - BACKGROUND: Venography is currently used to assess the incidence of deep vein thrombosis (DVT) in dose-finding and confirmatory trials of new antithrombotic agents. Centrally adjudicated, complete compression ultrasound (CCUS) could be a non-invasive alternative to venography. OBJECTIVES: A substudy of two, similarly designed, phase IIb trials of a novel, oral anticoagulant for the prevention of venous thromboembolism after elective hip or knee arthroplasty was undertaken to validate CCUS against venography. PATIENTS/METHODS: Patients received study drugs until mandatory, bilateral venography was performed 7 +/- 2 days after surgery. CCUS was performed within 24 h after venography by sonographers blinded to the venography result. Sonographers were trained and certified for the standardized examination and documentation procedure. Venograms and sonograms were adjudicated centrally at different sites by two independent readers; discrepancies between readers were resolved by consensus. RESULTS: A total of 1104 matching pairs of evaluable venograms and sonograms were obtained from the participants of the two trials (n = 1435): 19% of venograms and 20% of sonograms were not evaluable. The observed frequency of any DVT was 18.9% with venography and 11.5% with CCUS. Sensitivity of CCUS compared with venography was 31.1% for any DVT (95% confidence interval 23.4, 38.9), 21.0% (2.7, 39.4) for proximal DVT, and 30.8% (23.1, 38.6) for distal DVT. The figures for specificity were 93.0% (91.0, 95.1), 98.7% (98.0, 99.5), and 93.3% (91.5, 95.3), respectively. CONCLUSIONS: Based on these results, centrally adjudicated CCUS will be unable to replace venography for DVT screening early after major orthopaedic surgery in studies evaluating anticoagulant drugs. PMID- 17419764 TI - Bacillus anthracis anthrolysin O and three phospholipases C are functionally redundant in a murine model of inhalation anthrax. AB - Although traditionally considered to be an extracellular pathogen, Bacillus anthracis has a brief intracellular step to initiate anthrax. At the onset of infection, B. anthracis must withstand the bactericidal activities of the macrophage. Recently, three phospholipases C (PLCs) were shown to contribute to macrophage-associated growth of B. anthracis by presumably aiding in the escape of the bacterium from phagocytic vacuoles following phagocytosis. However, in the absence of all three PLCs, vegetative bacilli were still observed growing in association with the macrophage, albeit to a lesser extent, implicating that additional factors are involved in this process. In this study, the contributions of the previously identified cholesterol-dependent cytolysin anthrolysin O (ALO) to B. anthracis pathogenesis were investigated following challenges of bone marrow-derived macrophages and intratracheal inoculations of mice. Disruption of ALO alone yielded no differences in virulence in mice. However, combinatorial deletions of ALO with the three PLCs resulted in attenuation in both tissue culture and murine challenges, suggesting that these toxins may have overlapping roles in anthrax pathogenesis. PMID- 17419765 TI - Mutational analysis of NADH-binding residues in triphenylmethane reductase from Citrobacter sp. strain KCTC 18061P. AB - Triphenylmethane reductase (TMR) catalyzes the NADH-dependent reduction of triphenylmethane dyes. Sequence alignment revealed a region with a conserved GXXGXXG motif near its N-terminus, which corresponds to a conserved structural motif of known dinucleotide-binding proteins. To verify whether some of these glycine residues are important for the enzyme catalysis, these three glycine residues (Gly-7, Gly-10 and Gly-13) were individually replaced by alanine using site-directed mutagenesis. The secondary structures of these mutants, as measured by circular dichroism spectroscopy, did not show remarkable differences as compared with the wild type. The V(max)/K(m) values of mutants G7A and G13A for both Basic fuchsin and NADH were increased about three and twofold over that of the wild type, respectively, whereas the V(max)/K(m) value of mutant G10A were decreased about sixfold. These results suggest that these three glycine residues are involved in the interaction with both substrate and cofactor for the catalytic activity of TMR. PMID- 17419766 TI - Genotypic comparison of five isolates of Rickettsia prowazekii by multilocus sequence typing. AB - Genetic traits of five Rickettsia prowazekii isolates, including the first from Africa and North America, and representatives from human and flying squirrels were compared using multilocus sequence typing. Four rickettsial genes encoding 17 kDa genus-common antigen (17 kDa gene), citrate synthase (gltA), OmpB immunodominant antigen (ompB) and 120 kDa cytoplasmic antigen (sca4) were examined. Sequence identities of 17 kDa gene and gltA were 100% among the isolates. Limited sequence diversity of ompB (0.02-0.11%) and sca4 (0.03-0.20%) was enough to distinguish the isolates, and evaluation of the combined four genes provided a method to easily differentiate R. prowazekii from other rickettsiae. PMID- 17419767 TI - Mass spectrometric analysis of lipopeptides from Bacillus strains isolated from diverse geographical locations. AB - Matrix-assisted laser desorption/ionization time-of-flight MS (MALDI-TOF MS) has been applied to characterize lipopeptide biomarkers from 54 different strains of Bacillis from most taxa within the Bacillis subtilis-Bacillis licheniformis clade, isolated from seven geographic locations on five continents. Even the most narrowly defined taxa are diverse in terms of the lipopeptide profiles. Many strains produce previously identified compounds with known antimicrobial properties (e.g. polymyxins and bacitracins), whereas other compounds represent novel classes that were hitherto unknown. Of particular interest is the novel 942/958 Da biomarkers produced by B. s. spizizeni desert strains and several type strains. PMID- 17419768 TI - ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. AB - Recent progress in elucidating the role of the icaADBC-encoded polysaccharide intercellular adhesin (PIA) or polymeric N-acetyl-glucosamine (PNAG) in staphylococcal biofilm development has in turn contributed significantly to our understanding of the pathogenesis of device-related infections. Nevertheless, our understanding of how the ica locus and PIA/PNAG biosynthesis are regulated is far from complete and many questions remain. Moreover, beyond ica, evidence is now emerging for the existence of ica-independent biofilm mechanisms in both Staphylococcus aureus and Staphylococcus epidermidis. Teichoic acids, which are a major carbohydrate component of the S. epidermidis biofilm matrix and the major cell wall autolysin, play an important role in the primary attachment phase of biofilm development, whereas the cell surface biofilm-associated protein and accumulation-associated protein are capable of mediating intercellular accumulation. These findings raise the exciting prospect that other surface proteins, which typically function as antigenic determinants or in binding to extracellular matrix proteins, may also act as biofilm adhesins. Given the impressive array of surface proteins expressed by S. aureus and S. epidermidis, future research into their potential role in biofilm development either independent of PIA/PNAG or in cooperation with PIA/PNAG will be of particular interest. PMID- 17419769 TI - Reprogramming Hansenula polymorpha for penicillin production: expression of the Penicillium chrysogenum pcl gene. AB - We aim to introduce the penicillin biosynthetic pathway into the methylotrophic yeast Hansenula polymorpha. To allow simultaneous expression of the multiple genes of the penicillin biosynthetic pathway, additional markers were required. To this end, we constructed a novel host-vector system based on methionine auxotrophy and the H. polymorpha MET6 gene, which encodes a putative cystathionine beta-lyase. With this new host-vector system, the Penicillium chrysogenum pcl gene, encoding peroxisomal phenylacetyl-CoA ligase (PCL), was expressed in H. polymorpha. PCL has a potential C-terminal peroxisomal targeting signal type 1 (PTS1). Our data demonstrate that a green fluorescent protein-PCL fusion protein has a dual location in the heterologous host in the cytosol and in peroxisomes. Mutation of the PTS1 of PCL (SKI-COOH) to SKL-COOH restored sorting of the fusion protein to peroxisomes only. Additionally, we demonstrate that peroxisomal PCL-SKL produced in H. polymorpha displays normal enzymatic activities. PMID- 17419770 TI - Expression of the alpha3/beta1 isoform of human Na,K-ATPase in the methylotrophic yeast Pichia pastoris. AB - Na,K-ATPase is a crucial enzyme for ion homeostasis in human tissues. Different isozymes are produced by assembly of four alpha- and three beta-subunits. The expression of the alpha3/beta1 isozyme is confined to brain and heart. Its heterologous production has so far never been attempted in a lower eukaryote. In this work we explored whether the methylotrophic yeast Pichia pastoris is capable of expressing the alpha3/beta1 isoform of human Na,K-ATPase. cDNAs encoding the alpha(3) and the beta(1)-subunits were cloned under the control of the inducible promoter of Pichia pastoris alcohol oxidase 1. Pichia pastoris could express the single alpha3- and beta1-subunits and even coexpress them after methanol induction. beta1-subunit was produced as a major 44-kDa glycosylated polypeptide and alpha3 as a 110-kDa unglycosylated polypeptide. Expression at the plasma membrane was limited in shaking flask cultures but by cultivating P. pastoris cells in a fermenter there was a 10-fold increase of the number of ouabain binding sites per cell. The exported enzyme was estimated to be about 0.230 mg L( 1) at the end of a bioreactor run. Na,K-ATPase proved active and the dissociation constant of the recombinant enzyme-ouabain interaction was determined. PMID- 17419771 TI - Organelle association visualized by three-dimensional ultrastructural imaging of the yeast cell. AB - This study was aimed at a better understanding of organelle organization in the yeast Saccharomyces cerevisiae with special emphasis on the interaction and physical association of organelles. For this purpose, a computer aided method was employed to generate three-dimensional ultrastructural reconstructions of chemically and cryofixed yeast cells. This approach showed at a high level of resolution that yeast cells were densely packed with organelles that had a strong tendency to associate at a distance of <30 nm. The methods employed here also allowed us to measure the total surface area and volume of organelles, the number of associations between organelles, and the ratio of associations between organelles per surface area. In general, the degree of organelle associations was found to be much higher in chemically fixed cells than in cryofixed cells, with endoplasmic reticulum/plasma membrane, endoplasmic reticulum/mitochondria and lipid particles/nuclei being the most prominent pairs of associated fractions. In cryofixed cells, similar preferences for organelle association were seen, although at lower frequency. The occurrence of specific organelle associations is believed to be important for intracellular translocation and communication. Membrane contact as a possible means of interorganelle transport of cellular components, especially of lipids, is discussed. PMID- 17419772 TI - Redirection of peroxisomal alcohol oxidase of Hansenula polymorpha to the secretory pathway. AB - We report on the rerouting of peroxisomal alcohol oxidase (AO) to the secretory pathway of Hansenula polymorpha. Using the leader sequence of the Saccharomyces cerevisiae mating factor alpha (MFalpha) as sorting signal, AO was correctly sorted to the endoplasmic reticulum (ER), which strongly proliferated in these cells. The MFalpha presequence, but not the prosequence, was cleaved from the protein. AO protein was present in the ER as monomers that lacked FAD, and hence was enzymatically inactive. Furthermore, the recombinant AO protein was subject to gradual degradation, possibly because the protein did not fold properly. However, when the S. cerevisiae invertase signal sequence (ISS) was used, secretion of AO protein was observed in conjunction with bulk of the protein being localized to the ER. The amount of secreted AO protein increased with increasing copy numbers of the AO expression cassette integrated into the genome. The secreted AO protein was correctly processed and displayed enzyme activity. PMID- 17419773 TI - An ectophosphatase activity in Candida parapsilosis influences the interaction of fungi with epithelial cells. AB - This study describes the biochemical characterization of a phosphatase activity present on the cell surface of Candida parapsilosis, a common cause of candidemia. Intact yeasts hydrolyzed p-nitrophenylphosphate to p-nitrophenol at a rate of 24.30+/-2.63 nmol p-nitrophenol h(-1) 10(-7) cells. The cell wall distribution of the Ca. parapsilosis enzyme was demonstrated by transmission electron microscopy. The duration of incubation of the yeast cells with the substrate and cell density influenced enzyme activity linearly. Values of V(max) and apparent K(m) for p-nitrophenylphosphate hydrolysis were 26.80+/-1.13 nmol p nitrophenol h(-1) 10(-7) cells and 0.47+/-0.05 mM p-nitrophenylphosphate, respectively. The ectophosphatase activity was strongly inhibited at high pH as well as by classical inhibitors of acid phosphatases, such as sodium orthovanadate, sodium molybdate, sodium fluoride, and inorganic phosphate, the final product of the reaction. Only the inhibition caused by sodium orthovanadate was irreversible. Different phophorylated amino acids were used as substrates for the Ca. parapsilosis ectoenzyme, and the highest rate of phosphate hydrolysis was achieved using phosphotyrosine. A direct relationship between ectophosphatase activity and adhesion to host cells was established. In these assays, irreversible inhibition of enzyme activity resulted in decreased levels of yeast adhesion to epithelial cells. PMID- 17419774 TI - Transcriptional responses of Saccharomyces cerevisiae to preferred and nonpreferred nitrogen sources in glucose-limited chemostat cultures. AB - Aerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae grown with six different nitrogen sources were subjected to transcriptome analysis. The use of chemostats enabled an analysis of nitrogen-source-dependent transcriptional regulation at a fixed specific growth rate. A selection of preferred (ammonium and asparagine) and nonpreferred (leucine, phenylalanine, methionine and proline) nitrogen sources was investigated. For each nitrogen source, distinct sets of genes were induced or repressed relative to the other five nitrogen sources. In total, 131 such 'signature transcripts' were identified in this study. In addition to signature transcripts, genes were identified that showed a transcriptional coresponse to two or more of the six nitrogen sources. For example, 33 genes were transcriptionally upregulated in leucine-grown, phenylalanine-grown and methionine-grown cultures; this was partly attributed to the involvement of common enzymes in the dissimilation of these amino acids. In addition to specific transcriptional responses elicited by individual nitrogen sources, their impact on global regulatory mechanisms such as nitrogen catabolite repression (NCR) were monitored. NCR-sensitive gene expression in the chemostat cultures showed that ammonium and asparagine were 'rich' nitrogen sources. By this criterion, leucine, proline and methionine were 'poor' nitrogen sources, and phenylalanine showed an 'intermediate' NCR response. PMID- 17419775 TI - Peer review in nursing and midwifery: a literature review. AB - AIMS AND OBJECTIVES: The Clinical Education Project investigated clinical education in nursing and midwifery settings. The aim of this phase was to investigate and evaluate the processes and outcomes of clinical assessment of preregistration nurses and midwives, focusing on the assessment interview, and to evaluate the feasibility of introducing peer review of the clinical assessment interview in acute clinical settings. BACKGROUND: Peer review is common in many professional areas. The literature describes various applications of peer review and makes recommendations for its use. However, there is a shortage of studies investigating the use of peer review in nursing and midwifery education and practice. DESIGN: The project involved a systematic literature review and a qualitative exploratory study. This article describes the first part of the study: a systematic literature review of peer review. The second part of the study is reported elsewhere. METHODS: The systematic literature review investigated international articles written since 1994 that contained information on peer review in pre/post registration nursing and midwifery within higher education or practice. RESULTS: From the available literature, 52 specific initiatives were analysed. The majority of articles originated in America and involved nursing staff working in secondary care settings. Fifty-one articles had missing information varying from not stating the sample size to not including information about evaluations. CONCLUSIONS: The literature review found that whilst peer review is commonplace in nursing and midwifery practice, there is a lack of robust literature about its use. Relevance to clinical practice. Peer review in clinical settings such as nursing and midwifery can facilitate the sharing of good practice and personal and professional growth. It allows participants to learn from each other and gain insight into their development. PMID- 17419776 TI - Investigating the health profiles of Hong Kong Chinese: results of a cluster analysis. AB - AIM: The aim of this study was to identify the health profile of a group of Hong Kong Chinese adults. BACKGROUND: People were on the alert about their current health status and health has become the most valuable asset to many Hong Kong people. METHODS: A descriptive correlational survey was conducted during the period December 2003 - September 2004 in six districts in Hong Kong; 702 Chinese adults aged 18-64 completed a structured questionnaire. RESULTS: Cluster analysis revealed two profiles groups on their attitude towards social health, sexual behaviour, mental health, diet/nutrition, smoking/substance use, health knowledge and physical activity. CONCLUSIONS: Health data describing the health status of adults can be structured into a profile reflecting the characteristics of the target population under study. The findings showed that subjects in both clusters face two serious health issues: engage in little physical activity and possess poor knowledge about health. This finding does suggest that nurses can apply our understanding of these health profiles and to develop interventions that improve their health. RELEVANCE TO CLINICAL PRACTICE: This finding does suggest that nurses can apply our understanding of these health profiles and to develop interventions that improve their health. PMID- 17419777 TI - Spirituality and secularization: nursing and the sociology of religion. AB - AIM: The concept of spirituality is much discussed in the UK nursing literature, despite the fact that Britain is one of the most secular countries in the world, and steadily becoming more so. Here, I pose the following question: given this increasing secularization, what accounts for the current interest in spirituality among UK nurses? BACKGROUND: The literature on spirituality in nursing has blossomed in the last 10 years, and various attempts have been made to define 'spirituality', 'spiritual need' and 'spiritual care'. Most definitions distinguish between 'spirituality' and 'religion', acknowledging that the latter is more institutional, and theologically more restrictive, than the former; and they suggest that spirituality is universal, something which (unlike religion) all human beings share. METHOD: I draw on the sociology of religion - neglected, for the most part, in the nursing literature - to establish two main points. Firstly, that the UK and the USA are at opposite ends of the religion/secularity spectrum, implying that it is a mistake to assimilate USA and UK sources. Secondly, that the concept of spirituality, as currently understood, is of very recent origin, and is still 'under construction', having become separated from its associations with Christian piety and mysticism only since the 1980s. CONCLUSIONS: The extension of spirituality into secular domains is part of a professionalization project in nursing, a claim to jurisdiction over a newly invented sphere of work. For the time being, it remains an academic project (in the UK) as it is not one with which many clinicians identify. Relevance to clinical practice. What counts as 'spiritual need' or 'spiritual care' may not be the same in both countries, and UK clinicians are unlikely to welcome the role of surrogate chaplain, which their USA colleagues are apparently willing to embrace. PMID- 17419778 TI - A study on investigating unplanned readmission patterns. AB - AIM: To explain frequent hospital readmissions, this study aimed to determine whether definable subtypes exist within a cohort of subjects with chronic illness with regard to factors associated with a patient's readmission patterns and to compare whether these factors vary between subjects in groups with different profiles. RESEARCH METHOD: A descriptive correlational survey was conducted and data were collected by using a structured questionnaire. Seventy-four readmitted subjects were recruited in three general hospitals in Hong Kong. OUTCOME MEASURES: Five outcome variables were employed in the study: predisposing characteristic, need factors, health behaviour, health status or outcomes and enabling resources. RESULTS: A cluster analysis yielded two clusters. Each cluster represented a different profile of the sample on patient use of healthcare services. Cluster A consisted of 41.9% (n = 31) and Cluster B consisted of 58.1% (n = 43) of the patients. Cluster A patients, more of whom were male, were younger, more educated, had higher activities of daily living scores and fewer of them had received community nurse services than patients of Cluster B. Cluster A patients (32.3%) had more than one readmission record within 28 days than Cluster B patients (9.3%, p = 0.017). CONCLUSION: Our study shows that community nurse services can reduce the rate at which they are readmitted a second time. However, such services may have a positive effect only on a group of patients whose profile is similar to the patients in Cluster B and not for patients such as those in Cluster A. A clear profile may help healthcare policy makers make appropriate strategies to target a specific group of patients to reduce their readmission rates. RELEVANCE TO CLINICAL PRACTICE: The identification of risk for future healthcare use could enable better targeting of interventional strategies within these groups. The results of this study might provide hospital managers with a model to design specified interventions to reduce unplanned hospital readmissions for each profile group. PMID- 17419779 TI - A review of evidence-based practice, nursing research and reflection: levelling the hierarchy. AB - AIM: This paper examines the evidence-based practice movement, the hierarchy of evidence and the relationship between evidence-based practice and reflective practice. BACKGROUND: Evidence-based practice is equated with effective decision making, with avoidance of habitual practice and with enhanced clinical performance. The hierarchy of evidence has promoted randomized control trials as the most valid source of evidence. However, this is problematic for practitioners as randomized control trials overlook certain types of knowledge that, through the process of reflection, provide useful information for individualized and effective practice. METHOD: A literature search was undertaken using CINAHL, medline and Ovid electronic databases in early 2006. The search terms used were: evidence-based practice, research evidence, evidence for practice, qualitative research, reflective practice, reflection and evidence. Other sources included handpicking of books on evidence-based practice, reflection and research. Only material written in English was included. FINDINGS: The hierarchy of evidence that has promoted randomized control trials as the most valid form of evidence may actually impede the use of most effective treatment because of practical, political/ideological and epistemological contradictions and limitations. Furthermore, evidence-based practice appears to share very similar definitions, aims and procedures with reflective practice. Hence, it appears that the evidence based practice movement may benefit much more from the use of reflection on practice, rather than the use of the hierarchical structure of evidence. CONCLUSION: Evidence-based practice is necessary for nursing, but its' effective implementation may be hindered by the hierarchy of evidence. Furthermore, evidence-based practice and reflection are both processes that share very similar aims and procedures. Therefore, to enable the implementation of best evidence in practice, the hierarchy of evidence might need to be abandoned and reflection to become a core component of the evidence-based practice movement. RELEVANCE TO CLINICAL PRACTICE: Provides an elaborated analysis for clinical nurses on the definition and implementation of evidence in practice. PMID- 17419780 TI - Public Health Nurses and the delivery of quality nursing care in the community. AB - AIM: The aim of the study was to explore factors which impact on quality nursing care in the community from the Public Health Nurse's (PHN) perspective. BACKGROUND: Public Health Nursing has significantly evolved over the past few years with the delivery of quality nursing being a focus point. This study explores factors that impact upon the delivery of quality care in Public Health Nursing in Ireland. The findings provide an opportunity for an additional perspective to be included in the existing international findings and act as a starting point from which further research can be built. METHOD: A qualitative method using semi-structured interviews were conducted. Interviews were taped and content analysed. FINDINGS: Four main categories emefrged from the data, namely role change, components of quality nursing care, barriers to quality nursing care and the factors that facilitate the delivery of quality nursing care in the community. PHNs strive for evidence-based practice; they acknowledged their inability to achieve this and referred to factors that inhibited them from reaching their goal. CONCLUSION: Enhanced education for PHNs will equip them in the delivery of a quality service and have a positive impact on patient care. Better communication is required between PHNs, line managers and the multidisciplinary team. The delivery of community services need to be reviewed and developed further in accordance with the health strategy policy. RELEVANCE TO CLINICAL PRACTICE: This study has identified the evolution in clinical practice associated with the changing role and scope of Public Health Nursing. Clinical practice has evolved over time to incorporate societal change, technological advances and the delivery of an evidence-based service responsive to identified need. This study identified the presence of an increase in the specialist clinical work being undertaken as a result of new technological advances entering the community working environment. PMID- 17419781 TI - Adherence to clean intermittent self-catheterization procedures: determinants explored. AB - AIMS & OBJECTIVES: The aim of the current study was to explore factors that hinder or promote adherence to clean intermittent self-catheterization (CISC) procedures in adults. BACKGROUND: Clean intermittent self-catherization is associated with favourable patient outcomes, but adherence to the procedure is not addressed in the international literature. METHODS: Relevant factors were explored in two studies. The first study (n = 10) addressed mastery and short term adherence, whereas the second study (n = 20) addressed long-term adherence in these patients. Determinants of patient adherence were derived from pre structured interviews with patients, using a content-analysis procedure. RESULTS: A list of 16 determinants of mastery and short-term adherence and a list of 12 determinants of long-term adherence was found. Most of these determinants were found in both older (>or=65 years of age) and younger patients. However, five determinants of mastery and short-term adherence and six determinants of long term adherence were specific to patients under the age of 65. CONCLUSION: Our findings give a first insight into CISC adherence. General determinants of adherence relate to knowledge, complexity of the procedure, misconceptions, fears, shame, motivation and quality and continuity of professional care. Furthermore integrating CISC in everyday life can be difficult. In younger patients, availability of materials, physical impairments and resistance to a sickness role can further compromise adherence. RELEVANCE TO CLINICAL PRACTICE: Issues of knowledge, fears, motivation and potential psychological impact of performing CISC should be addressed prior to deciding on CISC and instructing patients. Follow-up care should be improved to include re-evaluations of skills, discussing adherence, integrating CISC in daily activities and general coping issues. PMID- 17419782 TI - Quality of life concerns and end-of-life care preferences of aged persons in long term care facilities. AB - AIMS AND OBJECTIVES: This study aimed at understanding quality of life (QOL) concerns and end-of-life care preferences of older people living in long-term care facilities in Hong Kong. BACKGROUND: Previous studies about the QOL of residents in long-term care facilities were often from a qualitative perspective, because it could yield an in-depth description of the experience. Quantitative studies in this area were not only scarce, but also less specific in capturing the concerns of this vulnerable group. Their forethought about their care planning was also overlooked. DESIGN AND METHOD: We successfully interviewed 287 older residents from 10 long-term care homes using the modified Quality-of-Life Concerns in the End of Life Questionnaire (QOLC-E) in a cross-sectional survey. RESULTS: The mean (SD) ages of the frail and non-frail groups were 83.8 (6.74) and 82.35 (5.82) respectively. The majority of them were female and widowed. Existential distress, value of life and food-related concerns were the least desirable QOL concerns in both groups. Considerable numbers were uncertain about their end-of-life care preferences and they preferred their physician to be their surrogate. CONCLUSION: The older residents had similar QOL concerns as patients with terminal or advanced diseases. Most of the older residents welcomed discussion about death and dying but had not planned for their end-of-life care and had a high regard for their physicians' authority. RELEVANCE TO CLINICAL PRACTICE: The results reveal the major QOL concerns among the older residents living in long-term care facilities. This study, which is an initial step in profiling the older people's end-of-life care preferences, also shed light on how to formulate advance care planning in long-term care facilities. PMID- 17419783 TI - Influences on parents' fever management: beliefs, experiences and information sources. AB - AIM: To identify parents' knowledge, beliefs, management and sources of information about fever management. BACKGROUND: Despite numerous studies exploring parents' management of childhood fever; negative beliefs about fever and overuse of antipyretics and health services for mild fevers and self-limiting viral illnesses continue to be reported. DESIGN: Qualitative design using semi structured interviews and discussions. METHOD: Fifteen metropolitan parents whose children were aged six months to five years, volunteered to participate in individual interviews or group discussions. Recruitment was through Playgroup Queensland's online newsletter and letters from two childcare centres to all parents. Verbatim and audio data were collected by an experienced moderator using a semi-structured interview guide. DATA ANALYSIS: Two transcripts were independently analysed by two researchers; categories, sub-headings and codes were independently developed, crosschecked and found comparable. Remaining transcripts were analysed using developed categories and codes. RESULTS: Fever, determined through behavioural changes, was perceived as 'good', a warning that something was wrong. High fever, reported as 38.0-39.1 degrees C, was considered harmful; it must be prevented or reduced irrespective of concerns about antipyretics. Positive febrile experiences reduced concern about fever. Negative experiences such as febrile convulsions, media reports of harm, not receiving a definitive diagnosis, inaccessibility to regular doctors and receiving conflicting information about fever management increased the concerns. Parents seek information about fever from multiple sources such as doctors, books and other parents. CONCLUSIONS: Parents' experiences with and information sources about fever and fever management influenced their knowledge, beliefs and practices. Positive experiences reduce concerns, health service usage and sometimes antipyretic usage. Negative experiences increase concerns, monitoring and antipyretic and health service usage. RELEVANCE TO CLINICAL PRACTICE: Health professionals need to update their fever management knowledge ensuring that it is based on the latest scientific knowledge. They must provide parents of young children with consistent, reliable information preferably before their first child's first febrile episode. PMID- 17419784 TI - Prevalence and associated factors of pneumonia in patients with vegetative state in Taiwan. AB - AIMS: The aim of this study was to investigate the prevalence rate and influencing factors of pneumonia associated with long-term tube feeding in special care units for patients with persistent vegetative states (PVS) in Taiwan. BACKGROUND: Pneumonia is a significant cause of morbidity, hospitalization and mortality in the nursing home population. Tube feeding has been found as a risk factor for the occurrence of pneumonia. METHODS: Two hundred sixty subjects were chosen from three hospital-based special care units for patients with PVS and 10 nursing facilities for persons in PVS in Taiwan. All subjects, who were diagnosed with PVS, received either financial aid for institutional care or were means-tested from The Bureau of Social Welfare of cities and counties in Taiwan. Data were collected through chart review and observations. RESULTS: The prevalence rate of pneumonia in nursing facilities for patients with PVS was 14.2%. The prevalence rate of tube-feeding in nursing facilities for PVS was 91.2%. The mean duration of tube-feeding was 73.21 SD 55.33 months. A total of 90.4% was fed with a nasogastric (NG) tube. Having a lower intake of food and fluids daily and having been institutionalized for a shorter period were three dominant factors associated with the occurrence of pneumonia. CONCLUSION: Research findings reveal that the incidence of pneumonia is higher in patients who do not receive adequate food and water. Continuing in service training to improve caregivers' knowledge and skill in providing care to patients in PVS and monitoring their skills in feeding is needed to decrease the occurrence of pneumonia in this population. RELEVANCE TO CLINICAL PRACTICE: Staff needs to be taught to monitor laboratory data and signs and symptoms of malnutrition and hydration deficit, and also be alert to early indicators of pneumonia in patients with PVS. PMID- 17419785 TI - Translation and validation of a Chinese version of the Self-Control Schedule in Chinese childbearing women. AB - AIMS: The aims of this study were to translate and evaluate the psychometric properties of the Chinese version of the Self-Control Schedule (C-SCS) in Chinese childbearing women. BACKGROUND: The SCS was developed to measure learned resourcefulness, which facilitates people's coping under adversity. Promoting learned resourcefulness in childbearing women facilitates positive adaptation to motherhood, which is essential to both maternal and child health. Thus, a reliable and valid C-SCS is crucial for the development of interventions to promote learned resourcefulness in Chinese childbearing women. DESIGN AND METHODS: In phase one, the SCS was translated into Chinese using the technique of translation and back-translation and was reviewed by an expert panel for cultural equivalence. In phase two, psychometric testing of the C-SCS was conducted by using a convenience sample of 360 childbearing women recruited from an antenatal clinic of a regional hospital. Four instruments including the C-SCS, General Self Efficacy Scale (GSE), Rosenberg's Self-Esteem Scale (RSES) and Edinburgh's Postnatal Depression Scale (EPDS) were administered to the participants. Ninety two women participated in the four-week retest. RESULTS: The C-SCS demonstrated good internal consistency (Cronbach's alpha = 0.83) and test-retest reliability (ICC = 0.91). Factor analysis revealed the presence of three factors supporting the conceptual dimension of the original instrument. Significant correlations with GSE (r = 0.48, p < 0.01), RSES (r = 0.41, p < 0.01) and EPDS (r = -0.46, p < 0.01) demonstrated good construct validity. CONCLUSION: The findings suggest that the C-SCS is a reliable and valid instrument for measuring learned resourcefulness in Chinese childbearing women. RELEVANCE TO CLINICAL PRACTICE: The C-SCS provides midwives with a useful tool in the evaluation of interventions designed to strengthen Chinese women's learned resourcefulness and promote maternal well-being during childbearing. PMID- 17419786 TI - Advanced nursing practice: an idea whose time has come. AB - AIMS AND OBJECTIVES: This paper aims to analyse the concept of advanced nursing practice critically and to demonstrate an appreciation of the development of the role of the advanced nurse practitioner in the Irish context. BACKGROUND: The concept of advanced nursing practice has existed in the US since the 1960s and in the UK since the early 1980s. The first signs of the advanced nurse practitioner began to emerge in Ireland in the late 1990s as a result of increasing demands and changes occurring within nursing and health care. Currently, there are 31 advanced nurse practitioners in the country, many of whom practice in the general setting. Strategies are in place to introduce advanced nursing practice into other nursing disciplines in the future. METHODS: A literature review was conducted to address the various issues inherent within this concept. RESULTS: There is a body of evidence indicating that advanced nursing practice has brought about an improvement in patient outcomes. In light of this, the advanced nurse practitioner has a valuable role to play in providing a beneficial contribution and filling a gap in healthcare services. CONCLUSION: This paper has demonstrated that nursing continues to be a dynamic and ever-changing phenomenon where the onus is firmly on the advanced nurse practitioner as expert, leader and collaborator, to capture the hearts and minds of nurses and healthcare workers, so as to challenge traditional values and transform clinical practice. Failure to do so may allow medicine or other interested parties to shape the future of nursing or even absorb it. RELEVANCE TO CLINICAL PRACTICE: Nurses working at an advanced level are striving to develop their expertise, initiate nurse-led services and practice, in collaboration with other professionals in an effort to provide the highest quality care to the patient. Although the idea of the advanced nurse practitioner is relatively new to nursing in Ireland, it is the result of an idea whose time has come and there is nothing more powerful than an idea whose time has come. PMID- 17419787 TI - From competence to capability: a study of nurse practitioners in clinical practice. AB - AIMS AND OBJECTIVES: This research aimed to understand the level and scope of practice of the nurse practitioner in Australia and New Zealand further using a capability framework. BACKGROUND: The original study, from which the present paper was developed, sought to identify competency standards for the extended role of the nurse practitioner in Australia and New Zealand. In doing so the researchers became aware that while competencies described many of the characteristics of the nurse practitioner they did not manage to tell the whole story. In a search of the literature, the concept of capability appeared to provide a potentially useful construct to describe the attributes of the nurse practitioner that went beyond competence. DESIGN: A secondary analysis of data obtained from interviews with nurse practitioners working in Australia and New Zealand was undertaken. These data had previously been obtained in a study to identify nurse practitioner competencies. The analysis described in this paper investigated whether or not the components of capability would adequately explain the characteristics of the nurse practitioner. METHODS: Fifteen nurse practitioners were interviewed from Australia and New Zealand. A secondary (deductive) analysis of interview data using capability as a theoretical framework was conducted. RESULTS: The analysis showed that capability and its dimensions is a useful model for describing the advanced level attributes of nurse practitioners. Thus, nurse practitioners described elements of their practice that involved: using their competences in novel and complex situations as well as the familiar; being creative and innovative; knowing how to learn; having a high level of self-efficacy; and working well in teams. CONCLUSIONS: This study suggests that both competence and capability need to be considered in understanding the complex role of the nurse practitioner. RELEVANCE TO CLINICAL PRACTICE: The dimensions of capability need to be considered in the education and evaluation of nurse practitioners. PMID- 17419788 TI - From monitoring physiological functions to using psychological strategies. Nurses' view of caring for the aneurysmal subarachnoid haemorrhage patient. AB - AIMS: The aims of this study were: (1) to describe nurses' views of the physical and supportive needs of patients who have suffered a subarachnoid haemorrhage (SAH), (2) to describe nurses' views of changes in social circumstances and (3) changes in the mental condition of patients after SAH. BACKGROUND: As patients with SAH are generally younger and predominantly female compared with other stroke groups they may have different needs of nursing support to facilitate adaptation. Caring for persons surviving stroke involves advanced nursing skills such as monitoring neurological functions in neurointensive care and providing physical care during rehabilitation. DESIGN: Explorative descriptive design. METHOD: Semi-structured interviews were performed with 18 nurses in neurointensive and rehabilitation care. A qualitative latent content analysis was conducted. RESULTS: Nurses viewed patients' need for support as a process ranging from highly advanced technological care to 'softer' more emotional care. However, shortages in the communication between neurointesive and rehabilitation nurses regarding this support were acknowledged. Changes in social circumstances and mental conditions were viewed both as obstacles and advantages regarding return to everyday life. Nurses also viewed that the characteristics of the group with SAH was not particularly different from the group with other types of stroke. CONCLUSIONS: Support to patients with SAH is viewed as a process carried out by nurses at neurointensive care units and rehabilitation units. Shortages in communication, regarding this support, were acknowledged. Obstacles and advantages with respect to returning to everyday life could apply to any stroke group, which could make it more difficult for nurses to detect the specific needs of patients with SAH. RELEVANCE TO CLINICAL PRACTICE: The communication between neurointensive nurses and rehabilitation nurses regarding support to patients with SAH is not satisfactory. Occasionally the specific needs of patients with SAH are not recognized. PMID- 17419789 TI - Participation of Icelandic nurses in smoking cessation counselling. AB - AIMS AND OBJECTIVES: The purpose of this study was to investigate the participation and knowledge of Icelandic nurses in smoking cessation counselling and to find barriers to smoking interventions by nurses. BACKGROUND: Research has shown that clinical intervention as brief as three minutes can substantially increase smoking cessation success. Several studies have revealed that majority of nurses agree that smoking cessation counselling is within their duties. However, the percentage of nurses who report advising and/or counselling patients remains low. DESIGN AND METHODS: An anonymous mail survey of all practicing nurses in Iceland was conducted in September 2004. A self-administered questionnaire was used, consisting of 74 questions in six sections: asking about smoking behaviour, to advise, to assess/assist/arrange, children and passive smoking, other questions and demographic questions. RESULTS: Of 2453 questionnaires, 868 complete questionnaires were returned, giving a 36% response rate. While the majority of nurses 'asked' about smoking behaviour, a minority 'advised' or 'assisted' their clients with smoking cessation. However, if the clients had no smoking-related symptoms, less than half of the nurses asked about smoking behaviour. Failure to ask and advise clients about the importance of smoking cessation and assisting with smoking cessation correlated (p < 0.001) with several factors, including: lack of time, insufficient knowledge or training and not considered as a part of daily duty. Nurses who smoked were less likely to advise against smoking (p < 0.05). CONCLUSIONS: Nurses frequently neglect to counsel clients about smoking cessation. Common barriers to the delivery of smoking cessation interventions by nurses include insufficient education and training in smoking cessation therapy. RELEVANCE TO CLINICAL PRACTICE: The number of patients with tobacco-related illnesses is increasing worldwide. Effective smoking cessation interventions by nurses have the enormous potential of reducing smoking prevalence and improve health. PMID- 17419790 TI - Injured road users' experience of care in the emergency department. AB - AIM: To describe the patients' perceptions of quality of care (QoC) in an emergency department (ED) and to analyse associations between patients' background characteristics and estimated QoC. BACKGROUND: Each year 1.4 out of every 100 inhabitants are injured in the traffic environment and receive care at the ED. No study has yet analysed different injured road users' perceived QoC, or how important they rate different caring dimensions. DESIGN: Cross-sectional study at the ED, in the University Hospital in Umea, Sweden. METHOD: A stratified consecutive sample of 166 car occupants, 200 cyclists and 199 pedestrians, aged 18-70 years, all injured in the traffic environment. Data were collected from medical records and from a mail survey using a short form of the Quality from the Patient's Perspective questionnaire, modified for ED use. The statistical methods used included Mann-Whitney's U-test, the Kruskal-Wallis test and multiple logistic regression. RESULTS: The perceived QoC and the subjective importance of the corresponding QoC dimensions were rated at the 'better' half of the rating scale, with no differences between the different road user categories. The most prominent factors associated with a positive perceived QoC rating were a short waiting time, moderate or serious injuries and high age as well as high educational level of the injury victim. For the subjective importance, a short waiting time was rated as the most important but slight differences were seen, related to education and sex. CONCLUSION: The association patterns between the areas of perceived reality and subjective importance indicated that expectations were higher than perceived QoC, suggesting that patients expected somewhat higher QoC than they received. RELEVANCE TO CLINICAL PRACTICE: Information on factors causing long waiting times, adapted to patients' age, sex and educational level, may reduce dissatisfaction among long waiting patients, especially among those with minor injuries. PMID- 17419791 TI - The role and potential contribution of clinical research nurses to clinical trials. AB - AIMS AND OBJECTIVES: This study explores the scope and potential contribution of the Clinical Research Nurse (CRN) role to clinical trials of a nursing-specific topic. BACKGROUND: Over the past two decades, there have been increases in the numbers of nurses working as CRNs because of the increasing global demand for clinical trials. CRNs can influence the quality of clinical trials but the scope and contribution of the role to clinical trials is not known. DESIGN: Qualitative focus group study. METHODS: A focus group interview was carried out with CRNs (n = 9) employed on a large, multi-centre (six NHS Trusts) randomized controlled trial of pressure area care. The focus group interview was recorded, alongside field notes of participant interactions and behaviours, and transcribed verbatim. Data were analysed for thematic content and process. FINDINGS: CRNs described their transition to a clinical research role. They reported a lack of confidence, role conflict as researcher and nurse, the challenges of gaining cooperation of clinical nursing staff to comply with trial protocols and difficulties maintaining their own motivation. CRNs provided their perceptions and observations of pressure area care and prevention. They identified areas of inadequate treatment, management and care, influenced by organizational and clinical aspects of care delivery. CONCLUSIONS: The study reveals challenges associated with training and management of CRNs. CRNs are usually associated with trial recruitment and data collection. This study highlights the additional contributions of CRNs for the study of topics specific to nursing as the result of their unique placement in the research centres as informal 'participant observers.' Such observations enhance understanding of the contexts being studied. RELEVANCE TO CLINICAL PRACTICE: These findings are relevant to the design and conduct of research studies of nursing care and practice and present ways for investigators to optimize the skills and knowledge of nurses working as CRNs. PMID- 17419792 TI - Innocent or culpable? Meanings that emergency department nurses ascribe to individual acts of violence. AB - AIMS AND OBJECTIVES: The purpose of the study was to explore the meaning(s) that emergency department nurses ascribe to acts of violence from patients, their family and friends and what impact these meaning(s) have upon how they respond to such acts. BACKGROUND: Violence in the health sector is of international concern. In high acuity areas such as emergency departments, nurses have an increased risk of violence. The literature further suggests that violence towards nurses in emergency departments is under-reported. DESIGN AND METHODS: This study was undertaken in 2005, at a regional Australian Emergency Department with 20 consenting registered nurses. Using an instrumental case study design, both qualitative and quantitative data were generated. Qualitative data were collected using participant observation, semi-structured interviews, informal field interviews and researcher journaling. Quantitative data of violent events were generated using a structured observational guide. Textual data were analysed thematically and numeric data were analysed using frequency counts. Mixed methods and concurrent data analysis contributed to the rigour of this study. FINDINGS: Emergency department nurses made judgments about the meaning of violent events according to three factors: (i) perceived personalization of the violence; (ii) presence of mitigating factors; and (iii) the reason for the presentation. The meanings that were ascribed to individual acts of violence informed the responses that nurses initiated. CONCLUSIONS; The findings show that violence towards emergency department nurses is interpreted in a more systematic and complex way than the current definitions of violence make possible. The meanings given to violence were contextually constructed and these ascribed meaning(s) and judgments informed the actions that the nurses took in response to both the act of violence and the agent of violence. Relevance to clinical practice. Understanding the meaning(s) of violence towards nurses contributes to the discussions surrounding why nurses under-report violence. Further, these findings bring insights into how nurses can and do, handle violence in the workplace. PMID- 17419793 TI - Sleep patterns and insomnia management in Korean-American older adult immigrants. AB - AIMS: This study aimed to describe sleep patterns and insomnia management in first generation Korean-American older adult immigrants. Specifically, this research examined differences in sleep interruption factors, use of sleep promotion aids, sleep characteristics and insomnia management between men and women who are first generation Korean-American older adult immigrants. BACKGROUND: Older adults feel that their sleep is shallow, interrupted frequently and is insufficient. If sleep changes are severe, it is difficult for older adults to maintain an awakened state during the day. DESIGN: This was a descriptive survey study. METHODS: The survey included a set of four questionnaires. All measures were self-administered. In the data analysis, descriptive statistics was used to analyse demographic characteristics. The chi squared test and t-test were used to examine the differences between men and women. RESULTS: Most subjects experienced sleep interruption (n = 43, 82.6%) and were not satisfied with their sleep (n = 42, 80.8%). A quarter of the subjects had experience with complementary/alternative therapies to manage insomnia, although 40 subjects (76.9%) wanted to use complementary/alternative therapies to manage their insomnia. CONCLUSIONS: The study shows that first generation Korean American older adult immigrant men and women report sleep interruptions and dissatisfaction with the quality of their sleep. Women were more likely in want of using complementary/alternative therapies for insomnia management than men. Women may have longed more for their traditional healthcare practices. The high degree of sleep disruption in this sample may relate to living in a different culture. RELEVANCE TO CLINICAL PRACTICE: Health professionals need to assess sleep patterns and consider an array of methods including complementary/alternative therapies to manage insomnia. PMID- 17419794 TI - Attitudes of nursing staff working with older people towards nutritional nursing care. AB - AIM: The aim of this study was to examine attitudes of the nursing staff in geriatric care towards factors of importance for nutritional nursing care. BACKGROUND: Studies show that nutritional risk assessment is seldom performed on older patients as routine and very few patients have a nutritional care plan. Patients in long-term care who are easy to feed are also found to be looked upon more positively than those with high feeding needs. METHODS: A total of 252 registered nurses and nurse aids working at geriatric rehabilitation and medical care clinics and resident homes participated in the study. Attitudes were examined using the Staff Attitudes to Nutritional Nursing Care Geriatric scale. The scale includes 18 items and was designed as a one to five-point Lickert-type scale. It gives a total score and five subscales representing the dimensions 'Norms', 'Habits', 'Assessment', 'Intervention' and 'Individualization'. A higher score indicates a more positive attitude. RESULTS: Of all nursing staff, 53% displayed a positive attitude towards factors of importance for nutritional nursing care and the rest displayed a neutral or negative attitude. The 'Intervention' dimension, dealing with nutritional problems and how to manage them, reflected the highest level of positive attitudes, which represents 71% of the nursing staff. The 'Norms' dimension had the lowest relative frequency of positive attitudes, 27%. The registered nurses held significantly more positive attitudes than the nurse aids did. CONCLUSIONS: Nutritional issues comprise an important and time-consuming responsibility in geriatric care; however, nursing staff do not show an unequivocal positive attitude regarding this responsibility. The consequences this entails for the older patient need to be examined further. Relevance to clinical practice. Nursing staff play an important role in caring for patients who are malnourished or at risk for malnutrition. Positive attitudes might hinder the development of undernourishment or the further worsening of an already undernourished patient's condition. PMID- 17419795 TI - Revisiting the concept of malnutrition in older people. AB - AIM: The aim of this study is to revisit the concept of malnutrition in older people, trace the new development and test the use of refined framework with empirical data. BACKGROUND: Malnutrition in older people is a common and significant problem worldwide. Continuing Chen's earlier work in 2001, a refinement was conducted and a prospective study was designed to test the use of this refined framework. DESIGN: A cohort study of 114 hospitalized older patients in Northern Taiwan. METHODS: The sample consists of 114 older patients aged 65 years and older, who were admitted for the cardiac and orthopaedic services at a tertiary 2300-bed hospital. From March to August 2004, assessed by one trained nurse, participants completed a structured face-to-face interview evaluating their age, visual/hearing impairments, oral health, cognitive status, comorbidities, medication use, social economic status, functional status, social support, depressive symptoms and nutritional status within 48 hours of admission. Participants who stayed >5 days were reassessed before discharge (n = 70). The data from admission were the main focus of this report. RESULTS: Regression analysis revealed that that more medication taken, female gender, lower functional status (beta = 0.34, P < 0.001) and higher depressive symptoms were independent predictors of poor nutritional status, with the full model accounting for 48.2% of the variance. The result is in-line with the original theoretical underpinnings and it suggests that this refined framework detailing sub-concepts and measurable indices appears to fit the empirical data and suitable for clinical use. CONCLUSION: The findings lend support to the use of this framework in managing malnutrition in older people. RELEVANCE TO CLINICAL PRACTICE: Nurses have an essential role in providing care for older people a framework like this would provide a road map guiding the intervention efforts. PMID- 17419796 TI - Developing an integrated career and competency framework for diabetes nursing. AB - AIMS AND OBJECTIVES: To describe the development of an integrated career and competency framework for diabetes nursing. BACKGROUND: The UK Nursing and Midwifery Council provides a definition of competence, but the terminology used in relation to the subject is often ambiguous and confusing. These concepts are explored in relation to nursing practice and the different approaches to competency framework development are described. To work alongside the Royal College of Nursing (RCN) and Skills for Health competency initiatives, a Diabetes Nursing Strategy Group representing nurses working in diabetes care was formed to oversee the development of an integrated career and competency framework for diabetes nursing. DESIGN: At the outset, the design was guided by the RCN Practice Development Team and employed qualitative methodology including the modified Delphi and nominal group technique. METHODS: A purposive sample of nurses representing all sectors and grades of staff involved in diabetes care was invited to workshops to undertake a values clarification exercise. Content analysis was performed to identify themes. Further workshops identified areas of specialist practice and competence statements were developed and refined in a series of consultations. RESULTS: Competence statements for a range of diabetes related areas were produced for nurses at the levels of unregistered practitioners, competent nurses, experience/proficient nurses, senior practitioners/expert nurses and consultant nurses. CONCLUSIONS: The description of the process of developing of the integrated career and competency framework should help other groups going through the same process. Relevance to clinical practice. In addition to helping groups identify a formula for the development of a competency framework, the framework itself is designed to provide a basis for educational programmes, personal career development and a tool for managers managing career progression within diabetes nursing. PMID- 17419797 TI - Quality of life for older people living in long-stay settings in Ireland. AB - AIM: The aim of this study was to explore the quality of life of older people living in long-stay care in Ireland from the perspectives of residents and staff. BACKGROUND: While the literature on quality of life is complex, several areas of agreement are evident. Quality of life is a multi-dimensional concept, which cannot be explained solely in medical terms and only makes sense if presented in an holistic context. Quality of life contains both subjective and objective elements; therefore, there is a need to take account of both when measuring the concept. METHOD: A mixed method research design was selected for this study. Three methods were employed; focus groups (n = 7) a quantitative survey of 526 long-stay facilities and qualitative interviews with 101 residents and 48 staff. RESULTS: The findings revealed four thematic domains of quality of life: care environment and ethos of care, personal identity, connectedness to family and community and activities and therapies. CONCLUSION: The study found that there were differences in resident experiences across care study sites and important differences in staffing provision, skill mix and physical environment between private and public facilities. Several factors were also found that either enhanced or diminished residents' potential to retain their connections and relationships with other people. Finally, residents were most likely to participate in activities which were designed to take into account their own particular interests. RELEVANCE TO CLINICAL PRACTICE: This study articulates domains of quality of life from the perspective of residents and staff and gives support to the claims that management and organizational structures within long stay care facilities matter for quality of life. Clear identification of what is important to older people is important as services need to be shaped around quality of life as much as quality of care, notwithstanding the close relationship between the two. PMID- 17419798 TI - Malnutrition in hip fracture patients: an intervention study. AB - AIMS: To investigate whether a nutritional intervention in older women and men with femoral neck fracture had an effect on postoperative complications during hospitalization and on nutritional status at a four-month follow-up. METHODS: The design was a randomized controlled trial. The present study sample consisted of 157 patients aged 70 years and above with femoral neck fracture. The nutritional intervention included, among other things, a nutritional journal to detect nutrition deficiencies and protein-enriched meals for at least four days postoperatively. Further, at least two nutritional and protein drinks were served each day during the whole hospitalization and other factors that would influence the patient's nutrition were also considered and dealt with. Postoperative complications were registered and patients were assessed using the Mini Nutritional Assessment (MNA) scale, including body mass index (BMI), on admission and at a four-month follow-up. RESULTS: Malnutrition was common and low MNA scores were associated with postoperative complications such as delirium and decubitus ulcers. There were significantly fewer days of delirium in the intervention group, seven patients in the intervention group developed decubitus ulcers vs. 14 patients in the control group and the total length of hospitalization was shorter. There were no detectable significant improvements regarding nutritional parameters between the intervention and the control group at the four-month follow-up but men improved their mean BMI, body weight and MNA scores in both the intervention and the control groups while women deteriorated in both groups. CONCLUSIONS: Malnutrition was common among older people with hip fractures admitted to hospital. The nutritional intervention might have contributed to the patients suffering fewer days with delirium, fewer decubitus ulcers and shorter hospitalization but did not improve the long-term nutritional status, at least not in women. RELEVANCE TO CLINICAL PRACTICE: This nutritional intervention, which was included in a multifactorial multidisciplinary intervention, is inexpensive and relatively easy to implement. It has significant effects on complications but no long-term effect on nutritional parameters, at least not in women. PMID- 17419799 TI - Interventions to enhance patient compliance with leg ulcer treatment: a review of the literature. AB - BACKGROUND AND AIMS: Non-compliance with compression therapy and with leg exercises and leg elevation is a common problem, often reported in patients with venous leg ulceration. Studies on compliance-enhancing interventions and the effectiveness of these interventions in patients with venous leg ulceration were reviewed. METHODS: MEDLINE, Cochrane, Embase and CINAHL were explored up to April 2005. Reference lists, wound care journals and conference proceedings were searched. Experts and manufacturers of compression systems were contacted. Studies were eligible if they included patients with venous or mixed leg ulcers and reported patient compliance outcome. Twenty studies were included. RESULTS: Most studies describe patient compliance as the extent to which the compression system was worn and/or the extent to which treatment regimen was followed. Self reporting was the most commonly used method of compliance assessment. There are indications that class III stockings for patients with venous ulcers enhance compliance compared with short stretch compression bandages. No real evidence is found that intermittent pneumatic compression systems improved compliance. There is no well-documented evidence that healthcare system interventions increase compliance. Educational programmes combining cognitive, behavioural and affective components were shown to have a positive effect on leg elevation, but not on compliance with compression therapy. CONCLUSION: The included studies have a lack of consistency in defining the standard and operationalization of compliance. Patient compliance plays an ancillary role in research. No study has been able to offer an acceptable and well-documented solution to the non-compliance problem. RELEVANCE TO CLINICAL PRACTICE: Research might focus on the development of comprehensive compliance-enhancing strategies. A stronger commitment of healthcare providers and society is needed to make progress in this area. The scope of nursing must be expanded to also include the problems experienced by patients with leg ulcers and the improvement of patient compliance. PMID- 17419800 TI - Leptin induces interleukin-1beta release from rat microglial cells through a caspase 1 independent mechanism. AB - Leptin regulates energy balance by suppressing appetite and increasing energy expenditure through actions in the hypothalamus. Recently we demonstrated that the effects of leptin are, at least in part, mediated by the release of interleukin (IL)-1beta in the brain. Microglia constitute the major source of IL 1beta in the brain but it is not known whether these cells express leptin receptors, or respond to leptin to produce IL-1beta. Using RT-PCR and immunocytochemistry, we demonstrate that primary rat microglial cells express the short (non-signalling) and long (signalling) isoforms of the leptin receptors (Ob R)s. Immunoassays performed on cell medium collected 24 h after leptin treatment (0.01-10 microg/mL) demonstrated a dose-dependent production and release of IL 1beta and its endogenously occurring receptor antagonist IL-1RA. In addition leptin-induced IL-1beta release occurs via a signal transducer and activator of transcription 3 (STAT3)-dependent mechanism. Western blot analysis demonstrated that leptin induced the synthesis of pro-IL-1beta in microglial cells and the release of mature 17 kDa isoform into the culture medium. Leptin-induced IL-1beta release was neither inhibited by the pan-caspase inhibitor BOC-D-FMK, nor by the caspase 1 inhibitor Ac-YVAD-CHO indicating that IL-1 cleavage is independent of caspase activity. These results confirm our earlier observations in vivo and demonstrate that microglia are an important source of IL-1beta in the brain in response to leptin. PMID- 17419801 TI - DNA methylation regulates tissue-specific expression of Shank3. AB - Tissue-specific gene expression can be controlled by epigenetic modifications such as DNA methylation. SHANK3, together with its homologues SHANK1 and SHANK2, has a central functional and structural role in excitatory synapses and is involved in the human chromosome 22q13 deletion syndrome. In this report, we show by DNA methylation analysis in lymphocytes, brain cortex, cerebellum and heart that the three SHANK genes possess several methylated CpG boxes, but only SHANK3 CpG islands are highly methylated in tissues where protein expression is low or absent and unmethylated where expression is present. SHANK3 protein expression is significantly reduced in hippocampal neurons after treatment with methionine, while HeLa cells become able to express SHANK3 after treatment with 5-Aza-2' deoxycytidine. Altogether, these data suggest the existence of a specific epigenetic control mechanism regulating SHANK3, but not SHANK1 and SHANK2, expression. PMID- 17419802 TI - Use of proteomics for the identification of novel drug targets in brain diseases. AB - In spite of the rapid advances in the development of the new proteomic technologies, there are, to date, relatively fewer studies aiming to explore the neuronal proteome. One of the reasons is the complexity of the brain, which presents high cellular heterogeneity and a unique subcellular compartmentalization. Therefore, tissue fractionation of the brain to enrich proteins of interest will reduce the complexity of the proteomics approach leading to the production of manageable and meaningful results. In this review, general considerations and strategies of proteomics, the advantages and challenges to exploring the neuronal proteome are described and summarized. In addition, this article presents an overview of recent advances of proteomic technologies and shows that proteomics can serve as a valuable tool to globally explore the changes in brain proteome during various disease states. Understanding the molecular basis of brain function will be extremely useful in identifying novel targets for the treatment of brain diseases. PMID- 17419803 TI - Irreversible blockade of sigma-1 receptors by haloperidol and its metabolites in guinea pig brain and SH-SY5Y human neuroblastoma cells. AB - We evaluated the effect of haloperidol (HP) and its metabolites on [(3)H](+) pentazocine binding to sigma(1) receptors in SH-SY5Y human neuroblastoma cells and guinea pig brain P(1), P(2) and P(3) subcellular fractions. Three days after a single i.p. injection in guinea pigs of HP (but not of other sigma(1) antagonists or (-)-sulpiride), [(3)H](+)-pentazocine binding to brain membranes was markedly decreased. Recovery of sigma(1) receptor density to steady state after HP-induced inactivation required more than 30 days. HP-metabolite II (reduced HP, 4-(4-chlorophenyl)-alpha-(4-fluorophenyl)-4-hydroxy-1 piperidinebutanol), but not HP-metabolite I (4-(4-chlorophenyl)-4 hydroxypiperidine), irreversibly blocked sigma(1) receptors in guinea pig brain homogenate and P(2) fraction in vitro. We found similar results in SH-SY5Y cells, which suggests that this process may also take place in humans. HP irreversibly inactivated sigma(1) receptors when it was incubated with brain homogenate and SH SY5Y cells, but not when incubated with P(2) fraction membranes, which suggests that HP is metabolized to inactivate sigma(1) receptors. Menadione, an inhibitor of the ketone reductase activity that leads to the production of HP-metabolite II, completely prevented HP-induced inactivation of sigma(1) receptors in brain homogenates. These results suggest that HP may irreversibly inactivate sigma(1) receptors in guinea pig and human cells, probably after metabolism to reduced HP. PMID- 17419804 TI - Fusion of diphtheria toxin and urotensin II produces a neurotoxin selective for cholinergic neurons in the rat mesopontine tegmentum. AB - Urotensin II is a neuropeptide first isolated from fish and later found in mammals: where it has potent cardiovascular, endocrine and behavioral effects. In rat brain the urotensin II receptor (UII-R) is predominately expressed in the cholinergic neurons of the pedunculopontine (PPTg) and laterodorsal tegmental nuclei. Typically, the function of the PPTg has been examined using excitotoxins, destroying both cholinergic and non-cholinergic neurons, which confounds interpretation. We took advantage of UII-R's unique expression profile, by combining UII with diphtheria toxin, to engineer a toxin specific for cholinergic neurons of the PPTg. In vitro, two different toxin constructs were shown to selectively activate UII-R (average EC50 approximately 30 nmol/L; calcium mobility assay) and to be 10,000-fold more toxic to UII-R expressing CHO cells, than wildtype cells (average LD50 approximately 2 nmol/L; cell viability). In vivo, pressure injection into the PPTg of rats, resulted in specific loss of choline transporter and NADPH diaphorase positive neurons known to express the UII-R. The lesions developed over time, resulting in the loss of over 80% of cholinergic neurons at 21 days, with little damage to surrounding neurons. This is the first highly selective molecular tool for the depletion of mesopontine cholinergic neurons. The toxin will help to functionally dissect the pedunculopontine and laterodorsal tegmental nuclei, and advance the understanding of the functions of these structures. PMID- 17419805 TI - Loss of hippocampal neuronal nitric oxide synthase contributes to the stress related deficit in learning and memory. AB - Nitric oxide (NO) has been involved in many pathophysiological brain processes. However, the exact role of NO in the cognitive deficit associated to chronic stress exposure has not been elucidated. In this study, we investigated the participation of hippocampal NO production and their regulation by protein kinase C (PKC) in the memory impairment induced in mice subjected to chronic mild stress model (CMS). CMS mice showed a poor learning performance in both open field and passive avoidance inhibitory task respect to control mice. Histological studies showed a morphological alteration in the hippocampus of CMS mice. On the other hand, chronic stress induced a diminished NO production by neuronal nitric oxide synthase (nNOS) correlated with an increment in gamma and zeta PKC isoenzymes. Partial restoration of nNOS activity was obtained after PKC activity blockade. NO production by inducible nitric oxide synthase isoform was not detected. The magnitude of oxidative stress, evaluated by reactive oxygen species production, after excitotoxic levels of NMDA was increased in hippocampus of CMS mice. Moreover, ROS formation was higher in the presence of nNOS inhibitor in both control and CMS mice. Finally, treatment of mice with nNOS inhibitors results in behavioural alterations similar to those observed in CMS animals. These findings suggest a novel role for nNOS showing protective activity against insults that trigger tissue toxicity leading to memory impairments. PMID- 17419806 TI - Rapid regulation of dopamine transporters by tyrosine kinases in rat neuronal preparations. AB - Termination of dopamine neurotransmission is primarily controlled by the plasma membrane-localized dopamine transporter. In this study, we investigated how this transporter is regulated by tyrosine kinases in neuronal preparations. In rat dorsal striatal synaptosomes, inhibition of tyrosine kinases by genistein or tyrphostin 23 resulted in a rapid (5-15 min), concentration-dependent decrease in [(3)H]dopamine uptake because of a reduction in maximal [(3)H]dopamine uptake velocity and dopamine transporter cell surface expression. The reduced transporter activity was associated with a decrease in phosphorylated p44/p42 mitogen-activated protein kinases. In primary rat mesencephalic neuronal cultures, the tyrosine kinase inhibitors similarly reduced [(3)H]dopamine uptake. When cultures were serum-deprived, acute activation of tyrosine kinase-coupled TrkB receptors by 100 ng/mL brain-derived neurotrophic factor significantly increased [(3)H]dopamine uptake; the effects were complex with increased maximal velocity but reduced affinity. The facilitatory effect of brain-derived neurotrophic factor on dopamine transporter activity depended on both the mitogen activated protein kinase and phosphatidylinositol 3-kinase pathways. Taken together, our results suggest that striatal dopamine transporter function and cell surface expression is constitutively up-regulated by tyrosine kinase activation and that brain-derived neurotrophic factor can mediate this type of rapid regulation. PMID- 17419807 TI - Major Cdk5-dependent phosphorylation sites of amphiphysin 1 are implicated in the regulation of the membrane binding and endocytosis. AB - Amphiphysin 1 (amph 1) is an endocytic protein enriched in the nerve terminals that functions in the clathrin-mediated endocytosis. It acts as membrane curvature sensor, a linker of clathrin coat proteins, and an enhancer of dynamin Guanosine Triphosphatase (GTPase) activity. Amph 1 undergoes phosphorylation by cyclin-dependent kinase 5 (Cdk5), at five phosphorylation sites, serine 262, 272, 276, 285, and threonine 310, as determined by mass spectrometry (MS). We show here that Cdk5-dependent phosphorylation of amph 1 is enhanced in the presence of lipid membranes. Analysis by tandem liquid chromatograph MS revealed that the phosphorylation occurs at two phosphorylation sites. The phosphorylation was markedly decreased by mutation either Ser276 or Ser285 of amph 1 to alanine (S276A and S285A). Furthermore, mutation of both sites (S276, 285A) completely eliminated the phosphorylation. Functional studies indicated that binding of amph 1 to lipid membrane was attenuated by Cdk5-dependent phosphorylation of wild type amph 1, but not of the S276, 285A form. Interestingly, endocytosis was increased in rat pheochromocytoma cells expressing amph 1 S276, 285A in comparison with wild type. These results suggest that Ser276 and Ser285 are regulatory Cdk5 phosphorylation sites of amph 1 in the lipid-bound state. Phosphorylation at these sites alters binding of amph 1 to lipid membranes, and may be an important regulatory aspect in the regulation of synaptic vesicle endocytosis. PMID- 17419808 TI - Inhibition of the myosin light chain kinase prevents hypoxia-induced blood-brain barrier disruption. AB - Increased mortality after stroke is associated with development of brain edema. The aim of the present study was to examine the contribution of endothelial myosin light chain (MLC) phosphorylation to hypoxia-induced blood-brain barrier (BBB) opening. Measurements of trans-endothelial electrical resistance (TEER) were performed to analyse BBB integrity in an in vitro co-culture model (bovine brain microvascular endothelial cells (BEC) and rat astrocytes). Brain fluid content was analysed in rats after stroke induction using a two-vein occlusion model. Dihydroethidium was used to monitor intracellular generation of reactive oxygen species (ROS) in BEC. MLC phosphorylation was detected using immunohistochemistry and immunoblot analysis. Hypoxia caused a decrease of TEER values by more than 40%, which was prevented by inhibition of the MLC-kinase (ML 7, 10 micromol/L). In addition, ML-7 significantly reduced the brain fluid content in vivo after stroke. The NAD(P)H-oxidase inhibitor apocynin (500 micromol/L) prevented the hypoxia-induced TEER decrease. Hypoxia-dependent ROS generation was completely abolished by apocynin. Furthermore, ML-7 and apocynin blocked hypoxia-dependent phosphorylation of MLC. Our data demonstrate that hypoxia causes a breakdown of the BBB in vitro and in vivo involving ROS and the contractile machinery. PMID- 17419809 TI - Proteomic analysis of gamma-butyrolactone-treated mouse thalamus reveals dysregulated proteins upon absence seizure. AB - Absence seizure has been of interest because the symptom is related to sensory processing. However, the mechanism that causes the disease is not understood yet. To better understand the molecular mechanism related to the disease progress at protein level, we performed proteomic studies using the thalamus of mice for which absence seizure was induced by gamma-butyrolactone (GBL). Differential proteome expression between GBL-treated mice and control mice was examined by fluorescence 2D difference gel electrophoresis (DIGE) at three different time points (5, 10, and 30 min) after GBL-administration. We identified 16 proteins differentially expressed by >1.4-fold at any of the three time points. All proteins besides the serine protease inhibitor EIA were down-regulated in absence seizure-induced mice. The down-regulated proteins can be classified into five groups by their biological functions: cytoskeleton rearrangement, neuroprotection, neurotransmitter secretion, calcium binding, and metabolism. The maximum level of change was reached by 10 min after GBL-treatment, with the expression level returning back to the original at 30 min when mice were awakened from absence seizure thereby demonstrating the proteomic response is reversible. Our results suggest that absence seizures are associated with restricted functional sets of proteins, whose down-regulation may interfere with general function of neuronal cells. PMID- 17419810 TI - Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures. AB - Nicotinic acetylcholine receptors play important roles in numerous cognitive processes as well as in several debilitating central nervous system (CNS) disorders. In order to fully elucidate the diverse roles of nicotinic acetylcholine receptors in CNS function and dysfunction, a detailed knowledge of their cellular and subcellular localizations is essential. To date, methods to precisely localize nicotinic acetylcholine receptors in the CNS have predominantly relied on the use of anti-receptor subunit antibodies. Although data obtained by immunohistology and immunoblotting are generally in accordance with ligand binding studies, some discrepancies remain, in particular with electrophysiological findings. In this context, nicotinic acetylcholine receptor subunit-deficient mice should be ideal tools for testing the specificity of subunit-directed antibodies. Here, we used standard protocols for immunohistochemistry and western blotting to examine the antibodies raised against the alpha3-, alpha4-, alpha7-, beta2-, and beta4-nicotinic acetylcholine receptor subunits on brain tissues of the respective knock-out mice. Unexpectedly, for each of the antibodies tested, immunoreactivity was the same in wild-type and knock-out mice. These data imply that, under commonly used conditions, these antibodies are not suited for immunolocalization. Thus, particular caution should be exerted with regards to the experimental approach used to visualize nicotinic acetylcholine receptors in the brain. PMID- 17419811 TI - Applicability of the glomerular size distribution coefficient in assessing human glomerular volume: the Weibel and Gomez method revisited. AB - Changes in glomerular volume (V(glom)) play an important role in the initiation and progression of various glomerulopathies. Estimation of V(glom) in the normal kidney provides baseline values for studies of glomerular hypertrophy in disease. The traditional model-based method of Weibel and Gomez is widely applied to estimate V(glom) in clinical biopsy specimens. Assumptions of glomerular size distribution and shape required by this method are potential sources of bias that have not been verified. We evaluated the applicability of the glomerular size distribution coefficient in estimating V(glom) in human kidneys. V(glom) of 720 non-sclerotic glomeruli in histologically normal kidneys of 24 males (20-69 years) was estimated by the unbiased disector/Cavalieri approach. Accurate glomerular diameters were calculated from Cavalieri estimates of V(glom) assuming glomerular sphericity. The coefficients of variation (CV) of glomerular diameters were compared with the corresponding values of the size distribution coefficient predicted by the Weibel and Gomez method. Mean (SD) glomerular diameter was 201 (28) mm (range 110-276 mm). The CV of glomerular diameter within each kidney ranged from 4.9 to 14.6%. Corresponding glomerular size distribution coefficients predicted by the formula of Weibel and Gomez ranged from 1.00 to just 1.03. The value of the size distribution coefficient required by the Weibel and Gomez technique when estimating V(glom) in normal human kidneys is remarkably constant. This is despite large variations in V(glom). Future studies should examine the extent of bias introduced by the glomerular shape assumptions of this method. PMID- 17419812 TI - Sensation and sexual arousal in circumcised and uncircumcised men. AB - INTRODUCTION: Research, theory, and popular belief all suggest that penile sensation is greater in the uncircumcised as compared with the circumcised man. However, research involving direct measurement of penile sensation has been undertaken only in sexually functional and dysfunctional groups, and as a correlate of sexual behavior. There are no reports of penile sensation in sexually aroused subjects, and it is not known how arousal affects sensation. In principle, this should be more closely related to actual sexual function. AIM: This study therefore compared genital and nongenital sensation as a function of sexual arousal in circumcised and uncircumcised men. METHODS: Twenty uncircumcised men and an equal number of age-matched circumcised participants underwent genital and nongenital sensory testing at baseline and in response to erotic and control stimulus films. Touch and pain thresholds were assessed on the penile shaft, the glans penis, and the volar surface of the forearm. Sexual arousal was assessed via thermal imaging of the penis. RESULTS: In response to the erotic stimulus, both groups evidenced a significant increase in penile temperature, which correlated highly with subjective reports of sexual arousal. Uncircumcised men had significantly lower penile temperature than circumcised men, and evidenced a larger increase in penile temperature with sexual arousal. No differences in genital sensitivity were found between the uncircumcised and circumcised groups. Uncircumcised men were less sensitive to touch on the forearm than circumcised men. A decrease in overall touch sensitivity was observed in both groups with exposure to the erotic film as compared with either baseline or control stimulus film conditions. No significant effect was found for pain sensitivity. CONCLUSION: These results do not support the hypothesized penile sensory differences associated with circumcision. However, group differences in penile temperature and sexual response were found. PMID- 17419813 TI - Is there a role for proteomics in Peyronie's disease? AB - INTRODUCTION: Peyronie's disease (PD) continues to be a major source of sexual dysfunction among the 3-9% of affected men. The challenge in treating PD is determining the natural history and clinical course for the individual patient. Currently, there exists no reliable means to predict whether a penile plaque of PD will progress, regress, or remain stable. This represents a significant deficiency in contemporary management, one that may be addressed with newer technologies such as proteomic profiling. AIM: This review assesses the potential use of protein alterations measured by various novel technologies, to predict progression, regression, or stabilization of PD in an affected individual. METHODS: A comprehensive literature review of the past decade in the field of gene profiling and protein expression of PD was performed. MAIN OUTCOME MEASURES: A critical analysis of the existing worldwide literature evaluating surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS or SELDI) and other proteonomic techniques. RESULTS: SELDI and other technologies can provide the clinician with innovative data indicating the presence of unique individual factors that act to suppress or promote the fibrotic process in PD. Determining the clinical implications of altered protein expression in an individual is not yet defined. CONCLUSIONS: The area of proteomics has begun to revolutionize the study of medicine in the postgenomic era, by allowing researchers to study the role that proteins play in health and disease. Applying this knowledge clinically has already led to innovative discoveries in early cancer detection in a number of malignancies, including prostate, ovarian, and bladder. Prior to the widespread use and acceptance of proteomic technology in PD, a critical assessment of its therapeutic and diagnostic value will be required. PMID- 17419814 TI - Exposure to an anabolic steroid changes female mice's sexual responses according to sex partner. AB - INTRODUCTION: Anabolic androgenic steroids (AAS) affect human female sexual behaviors. Animal models have been useful in uncovering the neural mechanisms governing changes in female sexual response upon AAS exposure. AIM: We quantify the sexual response of AAS-exposed gonadally intact female mice when paired with gonadally intact female or male pairs. METHODS: C57Bl/6 female mice were systemically exposed to the AAS 17alpha-methyltestosterone (7.5 mg/kg) for 17 days via a subcutaneous osmotic implant. On days 15-17, these females were allowed to mate with males or proestrus female partners in familiar and unfamiliar testing arenas for 10 minutes. MAIN OUTCOME MEASURES: The following behavioral responses were registered: parameters related to mounting behavior such as the frequency of mounts, attempts to mount, and the latency to the first mount, anogenital investigation, fights and escapes, rejections, pelvic thrusts, and lordotic responses. RESULTS: We found that males displayed a significant decrease in the frequency of mounts to AAS-exposed females, when compared with mating encounters with control females. We found no difference in the lordosis strength when control females were mounted by either a male or AAS-exposed females. However, females under androgen exposure attempted to mount control females, but not males, and their behavior was accompanied by significant increases in the number of fights, escapes, and rejections to the male. There were no differences between AAS-exposed females and males when the frequency of mounts and pelvic thrusts toward control females were compared. The lordotic quotient of control females was similar for either partner. CONCLUSIONS: Aside from showing a male-like pattern, AAS-exposed females displayed a higher frequency of anogenital investigations toward control females than males, and their latency to the first mount was as fast as that of males. Taken together, we conclude that the sex partner greatly influences the sexual response of AAS exposed female mice. PMID- 17419815 TI - Penile prosthesis implantation in cases of fibrosis: ultrasound-guided cavernotomy and sheathed trochar excavation. AB - INTRODUCTION: Implantation of a penile prosthesis into fibrosed corpora cavernosa is a difficult and risky procedure. Specialized instruments that assist safer and more efficient excavation include Otis Urethrotome and various cavernotomes, all of which operate underneath the tunica albuginea, out of sight. The blind use of such instruments can result in perforation of the tunica albuginea or injury to the urethra. AIM: This work describes the utility of ultrasonography for adding visual monitoring to any of the above-mentioned instruments, maintaining them in the mid-corpus cavernosum position to avoid perforation, and describes the application of alternative sheathed, sharp instruments that allow fast, efficient, and visually monitored drilling into fibrous tissue. MAIN OUTCOME MEASURES: Clinical outcome data were examined. METHODS: Surgery was performed on five cases with extensive fibrosis of the penis. Initial blunt dilatation by Hegar dilators faced considerable resistance. An ultrasound probe was applied to the ventral aspect of the penis. A laparoscopy sheath was advanced under ultrasound guidance up to the fibrous tissue. A sharp laparoscopy trochar was inserted through the sheath. Its tip was oriented in the mid-corpus cavernosum by longitudinal and transverse sonography sections, as it drilled into the fibrous tissue. Laparoscopy scissors were used in the same fashion to cut fibrous tissue lumps. After full excavation, penile prosthesis was implanted. RESULTS: All implants survived adequately. No complications occurred following implantation. Operative time ranged from 50 to 60 minutes. No difficulty was encountered at excavation. CONCLUSION: Ultrasound guidance can be a handy adjunct to any of the available techniques developed for excavating the fibrosed corpora cavernosa, with a possible decrease in difficulty and complication rate of the procedure. Utility of sheathed, sharp instruments guided by sonography is an alternative to the cavernotomes, allowing fast and efficient drilling into fibrous tissue. PMID- 17419816 TI - Sexual dysfunction in patients with chronic hand eczema in the Turkish population. AB - INTRODUCTION: Hand eczema can cause considerable psychosocial disorders, such as anxiety, depression, and difficulties at work, and it may also cause sexual dysfunction. AIM: The aim of this study was to investigate sexual function in patients with hand eczema and to find out whether concomitant depression has an additional negative effect on sexual function in these patients. MAIN OUTCOME MEASURES: Sexual functions were evaluated in hand eczema patients. METHODS: Ninety-one female (43 patients vs. 48 controls) and 79 male (45 patients vs. 34 controls) subjects were enrolled in the study. Hand eczema severity index was used to determine severity of hand eczema. The Female Sexual Function Index (FSFI) and the International Index of Erectile Function (IIEF) were used to assess sexual function. Quality of life was assessed with the Dermatology Life Quality Index. Diagnosis of depression was made based on the Structured Clinical Interview for the DSM-IV, while the Hamilton Depression Rate Scale was used for grading depression. RESULTS: Among 43 female subjects with hand eczema, 26 had depression (60.46%); of the 45 male patients, 11 had depression (24.44%). FSFI total score was found to be significantly decreased in female patients with both eczema and depression compared with controls (20.84 +/- 9.19 vs. 24.04 +/- 3.40, P < 0.05). FSFI total score was found to be significantly decreased in female patients with both eczema and depression compared with those without depression (20.84 +/- 9.19 vs. 22.23 +/- 5.82, P < 0.05). IIEF total score was also found to be significantly decreased in male patients with or without depression compared with controls (52.36 +/- 14.83 vs. 59.88 +/- 5.65 vs. 62.03 +/- 11.04, P < 0.05). CONCLUSIONS: The results of the study demonstrated that patients with hand eczema had sexual dysfunction, and concomitant depression had an additional negative effect on sexual dysfunction. Patients with hand eczema should be evaluated with regard to sexual function and depression to provide a better quality of life. PMID- 17419817 TI - Interrelationships among measures of premature ejaculation: the central role of perceived control. AB - INTRODUCTION: Domains of premature ejaculation (PE) include short intravaginal ejaculatory latency time (IELT), poor perceived control over ejaculation, decreased satisfaction with sexual intercourse, and personal distress and interpersonal difficulty related to ejaculation. How these measures interrelate is unknown. AIM: Here, we evaluated the interrelationships between these PE specific variables, applying cross-sectional data from a large U.S. observational study of men with PE. METHODS: We analyzed data from men with PE identified in a previously reported observational study. PE was diagnosed by experienced clinicians using the criteria specified in the Diagnostic and Statistical Manual of Mental Disorders, fourth edition, text revision. MAIN OUTCOME MEASURES: Subjects reported their stopwatch-measured IELT, perceived control over ejaculation, satisfaction with sexual intercourse, personal distress related to ejaculation, and interpersonal difficulty related to ejaculation. Relationships between variables were assessed using bivariate correlations, and the strength and significance of direct or indirect effects between variables were evaluated using a form of regression analysis known as path analysis. RESULTS: Bivariate Pearson correlation coefficients for all relationships were significant at the P 50% seizure reduction); two of these patients became seizure-free. Five out of seven patients with juvenile myoclonic epilepsy were responders. At the last follow-up visit, the patients had reduced the anti-epileptic drug (AED) usage from an average of 2.3 to 1.7 AED per patient (P = 0.0625). Two patients are currently being treated with VNS therapy only. Nine patients reported side effects, which were mostly mild and tended to diminish over time. CONCLUSION: Our results indicate that adjunctive VNS therapy is a favourable treatment option for patients with drug-resistant IGE. Rapid cycling seems worth trying in some of the non-responders. PMID- 17419831 TI - Which strategy should be applied? Design of a Norwegian epidemiological survey on chronic headache. AB - BACKGROUND: Large-scale population-based epidemiological surveys on chronic headache based on clinical interview by a physician are lacking. AIMS OF THE STUDY: To describe a method that aims to provide valid information on chronic headache. METHOD: A self-administered questionnaire including two questions about headache frequency within the last month and within the last year was used to screen for possible chronic headache. Respondents with self-reported headache more than half of the days were interviewed by neurological residents with experience in headache diagnostics. DISCUSSION: Previous studies have shown that a single question from a self-administered questionnaire can be used to screen for chronic headache. However, a precise headache diagnosis requires an interview by a physician experienced in headache diagnostics, as diagnoses extracted from questionnaires or provided by lay interviewers are not sufficiently precise. CONCLUSIONS: A combination of a screening questionnaire and a clinical interview by a physician is likely to be a cost-effective method to conduct an epidemiological survey on chronic headache. PMID- 17419832 TI - Clinical evolution of Kearns-Sayre syndrome with polyendocrinopathy and respiratory failure. AB - BACKGROUND: The triad of progressive external ophthalmoplegia, atypical retinal pigmentation and cardiac conduction defects characterizes Kearns-Sayre syndrome (KSS), which is most often caused by a single, large deletion of mitochondrial DNA. Endocrine disease appears to be more common in KSS than in other mitochondrial diseases. MATERIALS, METHODS AND RESULTS: A patient presenting with KSS developed Addison's disease, hypothyroidism and glucose intolerance. Thyroid peroxidase antibodies and adrenal 21-hydroxylase antibodies were identified. She developed acute respiratory failure requiring invasive ventilatory support, but improved and currently requires only non-invasive, nocturnal BiPAP treatment. DISCUSSION AND CONCLUSION: This case confirms the association of KSS and endocrine dysfunction. Our finding of autoantibodies to thyroid and adrenal glands distinguishes this patient from most other published cases and suggests a potential synergy between the two disease mechanisms. In addition, we demonstrate that respiratory failure can be a treatable event in this disease. PMID- 17419833 TI - Diaschisis after thalamic stroke: a comparison of metabolic and structural changes in a patient with amnesic syndrome. AB - INTRODUCTION: We present a patient with a left anteromedial thalamic lesion with an amnesic syndrome. The patient underwent neuropsychological testing, cerebrospinal fluid (CSF) analyses, magnetic resonance imaging (MRI) [T2, flair, and diffusion tensor imaging (DTI)] and [18F]-2-fluoro-deoxy-d-glucose positron emission tomography (FDG-PET) to assess indirect effects of thalamic lesions on cortical function. CASE REPORT: A 67-year-old right-handed woman was admitted to a university-based memory unit because of memory and concentration problems. Neuropsychological testing revealed dysfunction of episodic memory, semantic memory and working memory. General intellectual function and attention capacity were preserved. MRI revealed an anteromedial thalamic lesion in the left hemisphere. FDG-PET showed decreased uptake in the frontal, parietal and temporal lobes of the left hemisphere. Regions of interest (ROI) in white matter were selected and left and right hemispheres were compared. Fractional anisotropy (FA) in ROI representing thalamo-cortical connections were decreased in the left hemisphere when compared with the right. CONCLUSION: The results show the importance of a network that include the anterior and dorsomedian nuclei, which influence the activity in areas of the cortex responsible for memory processes. The imaging findings suggest that areas of cortical diaschisis after thalamic infarction correspond to areas affected by thalamo-cortical fibre loss as measured with FA. PMID- 17419834 TI - LRRK2 and Parkinson's disease in Norway. AB - OBJECTIVES: Mutations in the LRRK2 gene have been associated with both familial and sporadic late-onset Parkinson's disease. A large number of mutations in this gene have been identified; however, for many of these variants, the pathogenicity and relative frequency are unknown. Herein, we investigate the frequency of a number of recently identified LRRK2 mutations in Norway. METHODS: We genotyped eight putatively pathogenic LRRK2 mutations (R793M, R1067Q, I1371V, IVS31+3 A>G, M1869T, R1941H, T2356I and G2385R) in a series of 433 patients with Parkinson's disease and 587 controls from Norway. An intronic polymorphism previously reported to be associated with disease susceptibility was also examined (rs10506151). RESULTS: The Lrrk2 R793M substitution was found in two healthy individuals. No other LRRK2 mutations were identified in the Norwegian population, and furthermore no association was observed between rs10506151 and Parkinson's disease (P = 0.41). CONCLUSIONS: LRRK2 mutations other than the Lrrk2 G2019S mutation are rare in Norway. Our results indicate that the Lrrk2 R793M substitution is most likely a rare polymorphism. PMID- 17419835 TI - Cerebellar ataxia in the eastern and southern parts of Norway. AB - INTRODUCTION: The relative frequencies of different ataxias vary among different ethnic and geographic groups. The aim of this study was to examine patients with cerebellar ataxia and find the occurrence of autosomal dominant and recessive cerebellar ataxias in the population of the southern and eastern parts of Norway and estimate its prevalence. MATERIALS AND METHODS: Probands were systematically tested for spinocerebellar ataxia 1, 2, 3, 6 and Friedreich's ataxia. A total of 94 patients with ataxia were assessed. RESULTS: We registered 60 patients from 39 unrelated families with hereditary ataxias. One family with SCA2 (two patients), one family with Friedreich's ataxia (two patients), two patients heterozygote for Friedreich's ataxia and one metabolic ataxia were identified. CONCLUSIONS: We have few Friedreich's ataxia and SCA 1,2,3 and 6 in our population. Prevalence in Oslo County was estimated at 2.2/100,000 for autosomal recessive and 3.0/100,000 for autosomal dominant ataxia, respectively. PMID- 17419836 TI - WRINKLED1 specifies the regulatory action of LEAFY COTYLEDON2 towards fatty acid metabolism during seed maturation in Arabidopsis. AB - The WRINKLED1 (WRI1) transcription factor has been shown to play a role of the utmost importance during oil accumulation in maturing seeds of Arabidopsis thaliana. However, little is known about the regulatory processes involved. In this paper, comprehensive functional analyses of three new mutants corresponding to null alleles of wri1 confirm that the induction of WRI1 is a prerequisite for fatty acid synthesis and is important for normal embryo development. The strong expression of WRI1 specifically detected at the onset of the maturation phase in oil-accumulating tissues of A. thaliana seeds is fully consistent with this function. Complementation experiments carried out with various seed-specific promoters emphasized the importance of a tight regulation of WRI1 expression for proper oil accumulation, raising the question of the factors controlling WRI1 transcription. Interestingly, molecular and genetic analyses using an inducible system demonstrated that WRI1 is a target of LEAFY COTYLEDON2 and is necessary for the regulatory action of LEC2 towards fatty acid metabolism. In addition to this, quantitative RT-PCR experiments suggested that several genes encoding enzymes of late glycolysis, the fatty acid synthesis pathway, and the biotin and lipoic acid biosynthetic pathways are targets of WRI1. Taken together, these results indicate new relationships in the regulatory model for the control of oil synthesis in maturing A. thaliana seeds. In addition, they exemplify how metabolic and developmental processes affecting the developing embryo can be coordinated at the molecular level. PMID- 17419837 TI - Receptor-like protein kinase 2 (RPK 2) is a novel factor controlling anther development in Arabidopsis thaliana. AB - Receptor-like kinases (RLK) comprise a large gene family within the Arabidopsis genome and play important roles in plant growth and development as well as in hormone and stress responses. Here we report that a leucine-rich repeat receptor like kinase (LRR-RLK), RECEPTOR-LIKE PROTEIN KINASE2 (RPK2), is a key regulator of anther development in Arabidopsis. Two RPK2 T-DNA insertional mutants (rpk2-1 and rpk2-2) displayed enhanced shoot growth and male sterility due to defects in anther dehiscence and pollen maturation. The rpk2 anthers only developed three cell layers surrounding the male gametophyte: the middle layer was not differentiated from inner secondary parietal cells. Pollen mother cells in rpk2 anthers could undergo meiosis, but subsequent differentiation of microspores was inhibited by tapetum hypertrophy, with most resulting pollen grains exhibiting highly aggregated morphologies. The presence of tetrads and microspores in individual anthers was observed during microspore formation, indicating that the developmental homeostasis of rpk2 anther locules was disrupted. Anther locules were finally crushed without stomium breakage, a phenomenon that was possibly caused by inadequate thickening and lignification of the endothecium. Microarray analyses revealed that many genes encoding metabolic enzymes, including those involved in cell wall metabolism and lignin biosynthesis, were downregulated throughout anther development in rpk2 mutants. RPK2 mRNA was abundant in the tapetum of wild-type anthers during microspore maturation. These results suggest that RPK2 controls tapetal cell fate by triggering subsequent tapetum degradation, and that mutating RPK2 impairs normal pollen maturation and anther dehiscence due to disruption of key metabolic pathways. PMID- 17419838 TI - Environmental and hormonal regulation of the activity-dormancy cycle in the cambial meristem involves stage-specific modulation of transcriptional and metabolic networks. AB - We have performed transcript and metabolite profiling of isolated cambial meristem cells of the model tree aspen during the course of their activity dormancy cycle to better understand the environmental and hormonal regulation of this process in perennial plants. Considerable modulation of cambial transcriptome and metabolome occurs throughout the activity-dormancy cycle. However, in addition to transcription, post-transcriptional control is also an important regulatory mechanism as exemplified by the regulation of cell-cycle genes during the reactivation of cambial cell division in the spring. Genes related to cold hardiness display temporally distinct induction patterns in the autumn which could explain the step-wise development of cold hardiness. Factors other than low temperature regulate the induction of early cold hardiness-related genes whereas abscisic acid (ABA) could potentially regulate the induction of late cold hardiness-related genes in the autumn. Starch breakdown in the autumn appears to be regulated by the 'short day' signal and plays a key role in providing substrates for the production of energy, fatty acids and cryoprotectants. Catabolism of sucrose and fats provides energy during the early stages of reactivation in the spring, whereas the reducing equivalents are generated through activation of the pentose phosphate shunt. Modulation of gibberellin (GA) signaling and biosynthesis could play a key role in the regulation of cambial activity during the activity-dormancy cycle as suggested by the induction of PttRGA which encodes a negative regulator of growth in the autumn and that of a GA-20 oxidase, a key gibberellin biosynthesis gene during reactivation in spring. In summary, our data reveal the dynamics of transcriptional and metabolic networks and identify potential targets of environmental and hormonal signals in the regulation of the activity-dormancy cycle in cambial meristem. PMID- 17419839 TI - RNA silencing and its cell-to-cell spread during Arabidopsis embryogenesis. AB - In plants, post-transcriptional RNA silencing (PTGS) signals move beyond their sites of synthesis using channels called plasmodesmata (PD). However, a potential influence of PD permeability in silencing signal movement has not been addressed previously. PD connectivity and aperture are routinely determined by monitoring the cell-to-cell movement of protein tracers of different sizes. Here we compare protein and RNA silencing signal movement during Arabidopsis embryogenesis. The data suggest that the degree of PTGS signal movement correlates with the degree of PD aperture previously determined in distinct subregions of the developing Arabidopsis embryo. Silencing signals move more extensively in the hypocotyl and root compared with the area around the tips of the cotyledons, suggesting that the site of silencing signal initiation influences the extent of signal movement. In addition, we found a putative boundary for silencing spread just below the shoot apical meristem that blocks movement upward from the root and hypocotyl. Finally, as PD aperture affects protein movement in embryos, we compared the movement patterns of protein tracers versus the movement patterns of RNA silencing signals. The data reveal that silencing signal complexes move to a similar extent as soluble proteins between 27-54 kDa. PMID- 17419840 TI - AAP1 transports uncharged amino acids into roots of Arabidopsis. AB - Amino acids are available to plants in some soils in significant amounts, and plants frequently make use of these nitrogen sources. The goal of this study was to identify transporters involved in the uptake of amino acids into root cells. Based on the fact that high concentrations of amino acids inhibit plant growth, we hypothesized that mutants tolerating toxic levels of amino acids might be deficient in the uptake of amino acids from the environment. To test this hypothesis, we employed a forward genetic screen for Arabidopsis thaliana mutants tolerating toxic concentrations of amino acids in the media. We identified an Arabidopsis mutant that is deficient in the amino acid permease 1 (AAP1, At1g58360) and resistant to 10 mm phenylalanine and a range of other amino acids. The transporter was localized to the plasma membrane of root epidermal cells, root hairs, and throughout the root tip of Arabidopsis. Feeding experiments with [(14)C]-labeled neutral, acidic and basic amino acids showed significantly reduced uptake of amino acids in the mutant, underscoring that increased tolerance of aap1 to high levels of amino acids is coupled with reduced uptake by the root. The growth and uptake studies identified glutamate, histidine and neutral amino acids, including phenylalanine, as physiological substrates for AAP1, whereas aspartate, lysine and arginine are not. We also demonstrate that AAP1 imports amino acids into root cells when these are supplied at ecologically relevant concentrations. Together, our data indicate an important role of AAP1 for efficient use of nitrogen sources present in the rhizosphere. PMID- 17419841 TI - QQT proteins colocalize with microtubules and are essential for early embryo development in Arabidopsis. AB - During Arabidopsis embryogenesis, the control of division between daughter cells is critical for pattern formation. Two embryo-defective (emb) mutant lines named quatre-quart (qqt) were characterized by forward and reverse genetics. The terminal arrest of qqt1 and qqt2 embryos was at the octant stage, just prior to the round of periclinal divisions that establishes the dermatogen stage . Homozygous embryos of a weaker allele of qqt1 were able to divide further, resulting in aberrant periclinal divisions. These phenotypic analyses support an essential role of the QQT proteins in the correct formation of the tangential divisions. That an important proportion of qqt1 embryos were arrested prior to the octant stage indicated a more general role in cell division. The analysis of QQT1 and QQT2 genes revealed that they belong to a small subgroup of the large family encoding ATP/GTP binding proteins, and are widely conserved among plants, vertebrates and Archaea. We showed that QQT1 and QQT2 proteins interact with each other in a yeast two-hybrid system, and that QQT1 and QQT2 tagged by distinct fluorescent probes colocalize with microtubules during mitosis, in agreement with their potential role in cell division and their mutant phenotype. We propose that QQT1 and QQT2 proteins participate in the organization of microtubules during cell division, and that this function is essential for the correct development of the early embryo. PMID- 17419842 TI - Arbuscular mycorrhizal symbiosis is accompanied by local and systemic alterations in gene expression and an increase in disease resistance in the shoots. AB - In natural ecosystems, the roots of many plants exist in association with arbuscular mycorrhizal (AM) fungi, and the resulting symbiosis has profound effects on the plant. The most frequently documented response is an increase in phosphorus nutrition; however, other effects have been noted, including increased resistance to abiotic and biotic stresses. Here we used a 16,000-feature oligonucleotide array and real-time quantitative RT-PCR to explore transcriptional changes triggered in Medicago truncatula roots and shoots as a result of AM symbiosis. By controlling the experimental conditions, phosphorus related effects were minimized, and both local and systemic transcriptional responses to the AM fungus were revealed. The transcriptional response of the roots and shoots differed in both the magnitude of gene induction and the predicted functional categories of the mycorrhiza-regulated genes. In the roots, genes regulated in response to three different AM fungi were identified, and, through split-root experiments, an additional layer of regulation, in the colonized or non-colonized sections of the mycorrhizal root system, was uncovered. Transcript profiles of the shoots of mycorrhizal plants indicated the systemic induction of many genes predicted to be involved in stress or defense responses, and suggested that mycorrhizal plants might display enhanced disease resistance. Experimental evidence supports this prediction, and mycorrhizal M. truncatula plants showed increased resistance to a virulent bacterial pathogen, Xanthomonas campestris. Thus, the symbiosis is accompanied by a complex pattern of local and systemic changes in gene expression, including the induction of a functional defense response. PMID- 17419843 TI - Pathogen-associated molecular pattern recognition rather than development of tissue necrosis contributes to bacterial induction of systemic acquired resistance in Arabidopsis. AB - Systemic acquired resistance (SAR) is usually described as a phenomenon whereby localized inoculation with a necrotizing pathogen renders a plant more resistant to subsequent pathogen infection. Here we show that Pseudomonas syringae strains for which Arabidopsis thaliana represents a non-host plant systemically elevate resistance although the underlying interactions neither trigger a hypersensitive response nor cause necrotic disease symptoms. A similar enhancement of systemic resistance was observed when elicitor-active preparations of two typical bacterial pathogen-associated molecular patterns (PAMPs), flagellin and lipopolysaccharides (LPS), were applied in a localized manner. Several lines of evidence indicate that the observed systemic resistance responses are identical to SAR. Localized applications of non-adapted bacteria, flagellin or LPS elevate levels of the SAR regulatory metabolite salicylic acid (SA) and pathogenesis related (PR) gene expression not only in treated but also in distant leaves. All treatments also systemically increase expression of the SAR marker gene FLAVIN DEPENDENT MONOOXYGENASE 1. Further, a whole set of SAR-deficient Arabidopsis lines, including mutants in SA biosynthesis and signalling, are impaired in establishing the systemic resistance response triggered by non-host bacteria or PAMPs. We also show that the magnitude of defence reactions such as SA accumulation, PR gene expression or camalexin accumulation induced at sites of virulent or avirulent P. syringae inoculation but not the extent of tissue necrosis during these interactions determines the extent of SAR in distant leaves. Our data indicate that PAMPs significantly contribute to SAR initiation in Arabidopsis and that tissue necroses at inoculation sites are dispensable for SAR activation. PMID- 17419844 TI - Phytochrome coordinates Arabidopsis shoot and root development. AB - The phytochrome family of photoreceptors are potent regulators of plant development, affecting a broad range of responses throughout the plant life cycle, including hypocotyl elongation, leaf expansion and apical dominance. The plant hormone auxin has previously been linked to these phytochrome-mediated responses; however, these studies have not identified the molecular mechanisms that underpin such extensive phytochrome and auxin cross-talk. In this paper, we show that phytochrome regulates the emergence of lateral roots, at least partly by manipulating auxin distribution within the seedling. Thus, shoot-localized phytochrome is able to act over long distances, through manipulation of auxin, to regulate root development. This work reveals an important role for phytochrome as a coordinator of shoot and root development, and provides insights into how phytochrome is able to exert such a powerful effect on growth and development. This new link between phytochrome and auxin may go some way to explain the extensive overlap in responses mediated by these two developmental regulators. PMID- 17419845 TI - Differential regulation of closely related R2R3-MYB transcription factors controls flavonol accumulation in different parts of the Arabidopsis thaliana seedling. AB - The genes MYB11, MYB12 and MYB111 share significant structural similarity and form subgroup 7 of the Arabidopsis thaliana R2R3-MYB gene family. To determine the regulatory potential of these three transcription factors, we used a combination of genetic, functional genomics and metabolite analysis approaches. MYB11, MYB12 and MYB111 show a high degree of functional similarity and display very similar target gene specificity for several genes of flavonoid biosynthesis, including CHALCONE SYNTHASE, CHALCONE ISOMERASE, FLAVANONE 3-HYDROXYLASE and FLAVONOL SYNTHASE1. Seedlings of the triple mutant myb11 myb12 myb111, which genetically lack a complete subgroup of R2R3-MYB genes, do not form flavonols while the accumulation of anthocyanins is not affected. In developing seedlings, MYB11, MYB12 and MYB111 act in an additive manner due to their differential spatial activity; MYB12 controls flavonol biosynthesis mainly in the root, while MYB111 controls flavonol biosynthesis primarily in cotyledons. We identified and confirmed additional target genes of the R2R3-MYB subgroup 7 factors, including the UDP-glycosyltransferases UGT91A1 and UGT84A1, and we demonstrate that the accumulation of distinct and structurally identified flavonol glycosides in seedlings correlates with the expression domains of the different R2R3-MYB factors. Therefore, we refer to these genes as PFG1-3 for 'PRODUCTION OF FLAVONOL GLYCOSIDES'. PMID- 17419846 TI - Transcriptome profiling of maize anthers using genetic ablation to analyze pre meiotic and tapetal cell types. AB - Oligonucleotide arrays were used to profile gene expression in dissected maize anthers at four stages: after-anther initiation, at the rapid mitotic proliferation stage, pre-meiosis, and meiotic prophase I. Nearly 9200 sense and antisense transcripts were detected, with the most diverse transcriptome present at the pre-meiotic stage. Three male-sterile mutants lacking a range of normal cell types resulting from a temporal progression of anther failure were compared with fertile siblings at equivalent stages by transcription profiles. The msca1 mutant has the earliest visible phenotype, develops none of the normal anther cell types and exhibits the largest deviation from fertile siblings. The mac1 mutant has an excess of archesporial derivative cells and lacks a tapetum and middle layer, resulting in moderate transcriptional deviations. The ms23 mutant lacks a differentiated tapetum and shows the fewest differences from fertile anthers. By combining the data sets from the comparisons between individual sterile and fertile anthers, candidate genes predicted to play important roles during maize anther development were assigned to stages and to likely cell types. Comparative analyses with a data set of anther-specific genes from rice highlight remarkable quantitative similarities in gene expression between these two grasses. PMID- 17419847 TI - Cell division activity determines the magnitude of phosphate starvation responses in Arabidopsis. AB - Phosphate (P(i)) is a major limiting factor for plant growth. Plants respond to limiting P(i) supplies by inducing a suite of adaptive responses comprising altered growth behaviour, enhanced P(i) acquisition and reduced P(i) demand that together define a distinct physiological state. In P(i)-starved plants, continued root growth is required for P(i) acquisition from new sources, yet meristem activity consumes P(i). Therefore, we analysed the relationship between organ growth, phosphate starvation-responsive (PSR) gene expression and P(i) content in Arabidopsis thaliana under growth-promoting or inhibitory conditions. Induction of PSR gene expression after transfer of plants to P(i)-depleted conditions quantitatively reflects prior levels of P(i) acquisition, and hence is sensitive to the balance of P(i) supply and demand. When plants are P(i)-starved, enhanced root or shoot growth exacerbates, whereas growth inhibition suppresses, P(i) starvation responses, suggesting that the magnitude of organ growth activity specifies the level of P(i) demand. Inhibition of cell-cycle activity, but not of cell expansion or cell growth, reduces P(i) starvation-responsive gene expression. Thus, the level of cell-cycle activity specifies the magnitude of P(i) demand in P(i)-starved plants. We propose that cell-cycle activity is the ultimate arbiter for P(i) demand in growing organs, and that other factors that influence levels of PSR gene expression do so by affecting growth through modulation of meristem activity. PMID- 17419848 TI - Auxin, actin and growth of the Arabidopsis thaliana primary root. AB - To understand how auxin regulates root growth, we quantified cell division and elemental elongation, and examined actin organization in the primary root of Arabidopsis thaliana. In treatments for 48 h that inhibited root elongation rate by 50%, we find that auxins and auxin-transport inhibitors can be divided into two classes based on their effects on cell division, elongation and actin organization. Indole acetic acid (IAA), 1-naphthalene acetic acid (NAA) and tri iodobenzoic acid (TIBA) inhibit root growth primarily through reducing the length of the growth zone rather than the maximal rate of elemental elongation and they do not reduce cell production rate. These three compounds have little effect on the extent of filamentous actin, as imaged in living cells or by chemical fixation and immuno-cytochemistry, but tend to increase actin bundling. In contrast, 2,4-dichlorophenoxy-acetic acid (2,4-D) and naphthylphthalamic acid (NPA) inhibit root growth primarily by reducing cell production rate. These compounds remove actin and slow down cytoplasmic streaming, but do not lead to mislocalization of the auxin-efflux proteins, PIN1 or PIN2. The effects of 2,4-D and NPA were mimicked by the actin inhibitor, latrunculin B. The effects of these compounds on actin were also elicited by a 2 h treatment at higher concentration but were not seen in two mutants, eir1-1 and aux1-7, with deficient auxin transport. Our results show that IAA regulates the size of the root elongation zone whereas 2,4-D affects cell production and actin-dependent processes; and, further, that elemental elongation and localization of PINs are appreciably independent of actin. PMID- 17419849 TI - Overexpression of CRK13, an Arabidopsis cysteine-rich receptor-like kinase, results in enhanced resistance to Pseudomonas syringae. AB - Protein kinases play important roles in relaying information from perception of a signal to the effector genes in all organisms. Cysteine-rich receptor-like kinases (CRKs) constitute a sub-family of plant receptor-like kinases (RLKs) with more than 40 members that contain the novel C-X8-C-X2-C motif (DUF26) in the extracellular domains. Here we report molecular characterization of one member of this gene family, CRK13. Expression of this gene is induced more quickly and strongly in response to the avirulent compared with the virulent strains of Pseudomonas syringae, and peaks within 4 h after pathogen infection. In response to dexamethasone (DEX) treatment, plants expressing the CRK13 gene from a DEX inducible promoter exhibited all tested features of pathogen defense activation, including rapid tissue collapse, accumulation of high levels of several defense related gene transcripts including PR1, PR5 and ICS1, and accumulation of salicylic acid (SA). In addition, these plants suppressed growth of virulent pathogens by about 20-fold compared with the wild-type Col-0. CRK13-conferred pathogen resistance is salicylic acid-dependent. Gene expression analysis using custom cDNA microarrays revealed a remarkable overlap between the expression profiles of the plants overexpressing CRK13 and the plants treated with Pst DC3000 (avrRpm1). Our studies suggest that upregulation of CRK13 leads to hypersensitive response-associated cell death, and induces defense against pathogens by causing increased accumulation of salicylic acid. PMID- 17419850 TI - Abstracts of the 44th Conference of the German Society of Dermatology. April 25 28, 2007. Dresden, Germany. PMID- 17419852 TI - Subspecialization of radiation therapists in Australia and New Zealand. PMID- 17419853 TI - Exchanging dual-lumen central venous catheters: how I do it. AB - The management of a blocked long-term central venous catheter has traditionally been removal and re-siting, even when device failure is not associated with venous thrombosis or sepsis. It is not possible to 'railroad' a split dual-lumen central venous catheter down a long tortuous s.c. tunnel. Our exchange technique was designed to salvage the tunnel and venous access site in a long-term catheter that has had uncomplicated primary device failure. In this technique we divide the dual-lumen catheter and secure the venous access site and the s.c. tunnel with separate peel-away sheaths. The new catheter is then introduced in the conventional manner through the two peel-away sheaths, which are then removed. The key advantage of the technique is that it preserves one of the few central venous access sites available. This article describes exchange of an internal jugular catheter, but the same technique is applicable to subclavian catheters. PMID- 17419854 TI - Sarcomatous transformation in diaphyseal aclasis. AB - Multiple hereditary exostosis (or diaphyseal aclasis) is a condition characterized by the development of multiple osteochondromas. The tendency for malignant transformation into chondrosarcoma is well known. Malignancy typically arises from the cartilaginous cap of the osteochondroma. Radiographs supplemented by computed tomography have an important role in the diagnosis of this condition. Magnetic resonance imaging shows the features of sarcomatous change and aids in differentiating malignancy from pseudotumours. PMID- 17419855 TI - CT and MRI features in Bipolaris fungal sinusitis. AB - Bipolaris is an increasingly recognized cause of fungal sinusitis. Reports of imaging features are sparse. Our purpose was to review the imaging features in patients with Bipolaris fungal sinusitis. A review of our data showed seven patients with culture-proven Bipolaris fungal sinusitis. Computed tomography of the paranasal sinuses in all the patients and MRI in five patients were analysed for the location, nature, extent of the disease and density/signal characteristics on CT/MRI. The sphenoid and posterior ethmoid sinuses were most often involved (six of seven), followed by the anterior ethmoid sinus (five of seven), frontal sinus (four of seven) and maxillary sinus (three of seven) involvement. Five of seven cases had bilateral disease. Secretions were seen to fill the sinus and were expansile in nature in six of seven cases. Bony erosion was noted in all the patients. Air-fluid levels and bony sclerosis were rarely seen. Computed tomography showed central hyperdensity in all the cases. In the corresponding MR images (n = 5), the sinus contents appeared hyperintense on T1 weighted images and hypointense on T2-weighted images. Extension into the nasal cavity was found in six of seven cases. Five of seven cases had intracranial (extradural) spread. Intraorbital extension was seen in three of seven cases, with associated optic nerve compression in two. All the patients responded to surgical debridement, and systemic antifungal therapy was not required. Bipolaris fungal sinusitis typically presents with an allergic fungal sinusitis picture with expansile sinus opacification and bony erosions. There is central hyperdensity on CT scan, which appears hyperintense on T1-weighted and hypointense on T2-weighted MR images. PMID- 17419856 TI - Metastases to the breast: a review of 33 cases. AB - The aim of the study was to show the clinical and radiological manifestations of metastases to the breast (MB). From 1987 to 2006, 33 patients with MB were diagnosed at our institution. Their clinical and radiological features were retrospectively evaluated. Of the 33 cases, 31 presented as a palpable breast lump. On mammography, their findings were classified as follows: well circumscribed masses (11 cases), ill-circumscribed masses (five), focal asymmetric densities (one) and inflammatory skin changes (six). Mammograms were normal in six cases (all of them showed dense breast tissue). Four CT scans showed two well-circumscribed masses and two ill-circumscribed masses. Ultrasonography was available in 18 cases: hypoechoic lesions (15 cases) were more frequent than hyperechoic (one) or isoechoic lesions (two). The appearance on magnetic resonance was similar to primary breast cancer (one case). The most common primary tumours causing MB were haematological malignancies (nine cases) and melanomas (seven). Metastases to the breast showed a wide range of mammographic and ultrasonographic appearances, resembling both benign and malignant lesions. Any patient who presents with a breast lump with a history of cancer should undergo a core-needle biopsy in order to determine the histology of the lump. PMID- 17419857 TI - Determining the proportion of coronary segments assessable on 16-slice CT coronary angiography: a brief report. AB - Computed tomography coronary angiography (CT-CA) is becoming a popular non invasive coronary imaging method. We aimed to determine the proportion of coronary segments assessable on a 16-slice CT in comparison with conventional selective coronary angiography (SCA). We identified all patients who had both 16 slice CT-CA and recent SCA (less than 12 months) from March 2004 to July 2005. Two CT reporters blinded to SCA independently classified coronary segment assessability on CT-CA. A cardiologist blinded to CT findings classified assessability of coronary segments on SCA. Data were analysed using descriptive statistics and proportion of agreement. Ninety-five study pairs were included in the analysis. Of those, 1161 coronary segments were deemed assessable on SCA and 1103 segments (95%) were also assessable on CT-CA. Nonassessable segments on CT CA were predominantly in the distal segments and branches of coronary arteries. Reasons for nonassessability were small calibre (48.3%), motion artefacts (20.7%) and poorly reconstructed segments (22.4%). The 16-slice CT was able to assess a high proportion of but not all coronary segments. Nonassessable segments were predominantly distal segments or branches of coronary arteries. Motion artefacts due to heart-rate changes, small calibre and poorly reconstructed images were main causes of nonassessability on 16-slice CT-CA. PMID- 17419858 TI - Ureteroplasty with a cutting balloon: a novel approach to ureteric anastomotic strictures. AB - The Peripheral Cutting Balloon device was originally described to treat in-stent coronary artery stenosis. We aim to evaluate the use of this device in the management of ureteric anastomotic strictures. Four patients with radiologically proven ureteric anastomotic strictures were treated with the Peripheral Cutting Balloon Device using an antegrade approach. Follow up ranged between 2 and 12 months. All patients' initial procedure was successful, and all patients had improvement of symptoms. No patient showed evidence of biochemical deterioration of renal function, and no periprocedural complications occurred. One patient had restenosis as a result of invasive bladder cancer and required subsequent cystectomy. The Peripheral Cutting Balloon device is a novel instrument for the treatment of ureteric anastomotic strictures. Our experience shows promising short-term results but more follow up is required. PMID- 17419859 TI - Acute plain anterioposterior radiograph of the pelvis is not useful in detecting fractures of iliac wing and os sacrum: a prospective study of 73 patients using CT as gold standard. AB - Conflicting opinions exist to what extent interpreters of an acute anterioposterior (AP) radiograph of the pelvis underdiagnose pelvic injury. We have prospectively evaluated the usefulness of a plain AP radiograph of the pelvis in detecting injuries, using CT as the gold standard. Seventy-three haemodynamically stable blunt trauma patients, who had an AP radiograph of the pelvis and an abdominopelvic CT as part of their initial imaging evaluation, were included. There were 14 women and 59 men, with a mean age of 30 (3-61) years. Two senior radiologists, with special interest in CT examinations evaluating trauma, reviewed the radiographs and a month later the CT scans, blinded and independently. Usefulness index of the plain AP radiograph of the pelvis in detecting specific injuries was calculated using CT as the gold standard. A radiograph is regarded as useful if the usefulness index is 0.35 or more. The usefulness index for a plain AP radiograph of the pelvis for detecting a fracture of iliac wing and os sacrum was 0.25 and 0.01, respectively. An acute AP radiograph of the pelvis is not useful in detecting a fracture of iliac wing or os sacrum. PMID- 17419860 TI - Surface laser scanning to routinely produce casts for patient immobilization during radiotherapy. AB - Immobilization casts are used to reduce patient movement during the radiotherapy of head and neck and brain malignancies. Polyethylene-based casts are produced by first taking a Plaster of Paris 'negative' impression of the patient. A 'positive' mould is then made, which is used to vacuum form an immobilization cast. Taking the 'negative' cast can be messy, stressful for patients and labour intensive. Recently, lightweight hand-held laser surface scanners have become available. These allow an accurate 3-D representation of objects to be generated non-invasively. This technology has now been applied to the production of casts for radiotherapy. Each patient's face and head is digitized using the Polhemus FastSCAN (Polhemus, Colchester, VT, USA) scanner. The electronic data are transferred to a computer numerical controlled mill, where a positive impression is machined. The feasibility of the process was examined, the labour required and radiation therapists' satisfaction with aspects of the produced masks assessed. The scanner-based method of mask production was found to be simple, accurate and non-invasive. There was a reduction in radiation therapist labour required. Masks produced with the scanner-based method were reported to result in improved mask fitting, daily reproducibility, patient immobilization and patient comfort. PMID- 17419861 TI - Patient information preferences among breast and prostate cancer patients. AB - Preferences for information about their disease and treatment were collected from 392 patients who had been treated for either breast or prostate cancer an average of 2 years previously. Type of information that they had received, their ratings of its value to them and their preferred format for further information were examined. The most common and most preferred form of information was through doctor interview, followed by educational booklets. Prostate cancer patients preferred videotapes; breast cancer patients tended to prefer individualized approaches including a tour of the department. Effects of age, educational levels, occupational backgrounds and self-reports for anxiety and depression at the time of the survey and at time of diagnosis were analysed. Prostate cancer patients who were most severely depressed showed a preference for not receiving any information at all, perhaps reflecting a tendency towards withdrawal. PMID- 17419862 TI - Music therapy as a non-pharmacological anxiolytic for paediatric radiotherapy patients. AB - Outpatient radiotherapy treatment in the paediatric cancer patient can be a traumatic and an anxiety-provoking experience for both the patient and the family. Music therapy has been widely reported to have psychosocial, educational and physical benefits for the paediatric cancer patient. Using individual case reports, this paper shows the successful use of music therapy as a non pharmacological anxiolytic in the paediatric radiotherapy, outpatient waiting room setting, by providing the patient and the family with a means of communication, self-expression and creativity. PMID- 17419863 TI - Acquired spontaneous intercostal abdominal hernia: case report and a comprehensive review of the world literature. AB - Intercostal hernias develop most often as a result of a blunt or penetrating thoracoabdominal trauma. We know of no prior report of a spontaneously occuring intercostal hernia. This study presents a review of the published literature that deals with this uncommon phenomenon, along with a discussion of our patient's clinical presentation and imaging findings. PMID- 17419864 TI - Extramedullary haematopoiesis in thalassaemia major causing spinal cord compression. AB - A 33-year-old, homozygous beta-thalassaemic, Jehovah witness man presented with subacute spinal cord compression secondary to extramedullary haematopoiesis within the thoracic spinal canal. In this case, MRI showed characteristic features of extramedullary haematopoiesis, leading to an early diagnosis. PMID- 17419865 TI - Bilateral spontaneous adrenal haemorrhage complicating acute pancreatitis. AB - Bilateral adrenal haemorrhage is an event that mandates prompt diagnosis and treatment to prevent primary adrenocortical insufficiency and potential death. Presentation can be non-specific and incidentally diagnosed with imaging alone, primarily CT. We present a case of acute pancreatitis with spontaneous bilateral adrenal haemorrhage and briefly discuss imaging and treatment implications. PMID- 17419866 TI - Two cases of methanol poisoning: CT and MRI features. AB - Methanol poisoning in Australia is now very rare as methanol has been removed from methylated spirits. In acute intoxication methanol may result in a wide range of damage to the central nervous system. Few cases have been imaged with MRI. We present two cases and their striking neuroimaging findings with a discussion of the published work on methanol poisoning. PMID- 17419867 TI - Percutaneous retrieval of a chronic foreign body with both intravascular and extravascular components. AB - Removal of unwanted intravascular foreign body is a useful but infrequent procedure carried out by interventional radiologists. We study a patient who had a long guidewire left in her body following central venous catheter placement by a surgeon. The guidewire was later found in situ, with both intravascular and extravascular components in continuity. We successfully removed the guidewire without causing any complications. Standard interventional techniques, Amplatz gooseneck snare (Microvena, White Bear Lake, MN, USA) and 6-Fr Multipurpose catheter were used. PMID- 17419868 TI - Percutaneous fluoroscopic removal of a knotted Swan-Ganz catheter in a patient with a persistent left-sided superior vena cava. AB - Knotting of intravascular catheters is an uncommon but a well-recognized occurrence. The Swan-Ganz catheter (SGC) is the one that knots most commonly. A case of a knotted SGC is described in a patient with a persistent left-sided superior vena cava, and we propose that the presence of a left-sided superior vena cava is a risk factor for knot formation not previously reported. We review the published work on the risk factors for knot formation and on the techniques used to remove knotted SGC. We describe a technique using a gooseneck snare and Omni Flush catheter (Angiodynamics, Queensbury, NY, USA) to loosen and untie a knotted SGC. PMID- 17419869 TI - Prolonged survival after resection and radiotherapy for solitary brain metastases from non-small-cell lung cancer. AB - Selected patients with brain metastases from non-small-cell lung cancer benefit from aggressive treatment. This report describes three patients who developed solitary brain metastases after previous resection of primary adenocarcinoma of the lung. Each underwent surgical resection of their brain metastasis followed by cranial irradiation and remain disease free 10 or more years later. Two patients developed cognitive impairment approximately 8 years after treatment of their brain metastasis, which was felt to be due to their previous brain irradiation. Here we discuss the treatment of solitary brain metastasis, particularly the value of combined method approaches in selected patients and dose-volume considerations. PMID- 17419870 TI - Emergency transcatheter embolization of ruptured hepatocellular carcinomas with tortuous conventional or aberrant hepatic vascular anatomy, or parasitic supply. AB - Emergency transcatheter embolization is a well-recognized measure to manage patients with life-threatening haemoperitoneum due to spontaneous ruptured hepatocellular carcinoma. Traditional embolization technique is to embolize the proper hepatic artery or the segmental hepatic artery by femoral approach using gelfoam pledgets. From 1997 to 2004, in 19 out of 96 embolizations, the embolization technique had to be modified because of tortuous conventional or aberrant hepatic vascular anatomy or parasitic supply to achieve successful embolization. PMID- 17419871 TI - How accurate is the Light index for estimating pneumothorax size? AB - In Europe, the Light index is used for estimating the size of pneumothorax. It is based on the proportional relationship between the collapsed lung and the hemithorax. Other methods rely on absolute measurements on radiographs. The aim of this study was to compare the estimated size of pneumothoraces using Light index with that estimated using the CT volumetrics-derived Collins method. This was a secondary analysis of radiographs from a database of pneumothoraces treated conservatively. Radiographs were measured independently by two researchers according to the methods described by Light and Collins. Estimates of size derived by the two methods were compared using bias plot analysis techniques. The primary outcome of interest was agreement between pneumothorax size (as per cent) between the two methods. Sixty-one radiographs (27 patients) were analysed. The median age was 21 years and 76% were men. There was poor agreement between the methods. The average difference was 7.3% with the Light index underestimating the size compared with the Collins method. The 95% limits of agreement were wide (24% underestimation to 17% overestimation of size). The Light index as described does not accurately estimate the size of pneumothorax. New methods if accurate quantification of pneumothoraces is required clinically. PMID- 17419872 TI - Re: the ongoing importance of plain film interpretation. PMID- 17419873 TI - Re: ownership of medical images. PMID- 17419874 TI - Re: rectosigmoid cancer after radiotherapy for prostate cancer can be detected early and successfully treated. PMID- 17419875 TI - Genetic mapping of a new heart rate QTL on chromosome 8 of spontaneously hypertensive rats. AB - BACKGROUND: Tachycardia is commonly observed in hypertensive patients, predominantly mediated by regulatory mechanisms integrated within the autonomic nervous system. The genetic loci and genes associated with increased heart rate in hypertension, however, have not yet been identified. METHODS: An F2 intercross of Spontaneously Hypertensive Rats (SHR) x Brown Norway (BN) linkage analysis of quantitative trait loci mapping was utilized to identify candidate genes associated with an increased heart rate in arterial hypertension. RESULTS: Basal heart rate in SHR was higher compared to that of normotensive BN rats (365 +/- 3 vs. 314 +/- 6 bpm, p < 0.05 for SHR and BN, respectively). A total genome scan identified one quantitative trait locus in a 6.78 cM interval on rat chromosome 8 (8q22-q24) that was responsible for elevated heart rate. This interval contained 241 genes, of which 65 are known genes. CONCLUSION: Our data suggest that an influential genetic region located on the rat chromosome 8 contributes to the regulation of heart rate. Candidate genes that have previously been associated with tachycardia and/or hypertension were found within this QTL, strengthening our hypothesis that these genes are, potentially, associated with the increase in heart rate in a hypertension rat model. PMID- 17419876 TI - Fisher's combined p-value for detecting differentially expressed genes using Affymetrix expression arrays. AB - BACKGROUND: Currently, most tests of differential gene expression using Affymetrix expression array data are performed using expression summary values representing each probe set on a microarray. Recently testing methods have been proposed which incorporate probe level information. We propose a new approach that uses Fisher's method of combining evidence from multiple sources of information. Specifically, we combine p-values from probe level tests of significance. RESULTS: The combined p method and other competing methods were compared using three spike-in datasets (where probe sets corresponding differentially spiked transcripts are known) and array data from a biological study validated with qRT-PCR. Based on power and false discovery rates computed for the spike-in datasets, we demonstrate that the combined p method compares favorably with other methods. We find that probe level testing methods select many of the same genes as differentially expressed. We illustrate the use of the combined p method for diagnostic purposes using examples. CONCLUSION: Combined p is a promising alternative to existing methods of testing for differential gene expression. In addition, the combined p method is particularly well suited as a diagnostic tool for exploratory analysis of microarray data. PMID- 17419877 TI - Assembly of the Candida albicans genome into sixteen supercontigs aligned on the eight chromosomes. AB - BACKGROUND: The 10.9x genomic sequence of Candida albicans, the most important human fungal pathogen, was published in 2004. Assembly 19 consisted of 412 supercontigs, of which 266 were a haploid set, since this fungus is diploid and contains an extensive degree of heterozygosity but lacks a complete sexual cycle. However, sequences of specific chromosomes were not determined. RESULTS: Supercontigs from Assembly 19 (183, representing 98.4% of the sequence) were assigned to individual chromosomes purified by pulse-field gel electrophoresis and hybridized to DNA microarrays. Nine Assembly 19 supercontigs were found to contain markers from two different chromosomes. Assembly 21 contains the sequence of each of the eight chromosomes and was determined using a synteny analysis with preliminary versions of the Candida dubliniensis genome assembly, bioinformatics, a sequence tagged site (STS) map of overlapping fosmid clones, and an optical map. The orientation and order of the contigs on each chromosome, repeat regions too large to be covered by a sequence run, such as the ribosomal DNA cluster and the major repeat sequence, and telomere placement were determined using the STS map. Sequence gaps were closed by PCR and sequencing of the products. The overall assembly was compared to an optical map; this identified some misassembled contigs and gave a size estimate for each chromosome. CONCLUSION: Assembly 21 reveals an ancient chromosome fusion, a number of small internal duplications followed by inversions, and a subtelomeric arrangement, including a new gene family, the TLO genes. Correlations of position with relatedness of gene families imply a novel method of dispersion. The sequence of the individual chromosomes of C. albicans raises interesting biological questions about gene family creation and dispersion, subtelomere organization, and chromosome evolution. PMID- 17419878 TI - Selective prediction of interaction sites in protein structures with THEMATICS. AB - BACKGROUND: Methods are now available for the prediction of interaction sites in protein 3D structures. While many of these methods report high success rates for site prediction, often these predictions are not very selective and have low precision. Precision in site prediction is addressed using Theoretical Microscopic Titration Curves (THEMATICS), a simple computational method for the identification of active sites in enzymes. Recall and precision are measured and compared with other methods for the prediction of catalytic sites. RESULTS: Using a test set of 169 enzymes from the original Catalytic Residue Dataset (CatRes) it is shown that THEMATICS can deliver precise, localised site predictions. Furthermore, adjustment of the cut-off criteria can improve the recall rates for catalytic residues with only a small sacrifice in precision. Recall rates for CatRes/CSA annotated catalytic residues are 41.1%, 50.4%, and 54.2% for Z score cut-off values of 1.00, 0.99, and 0.98, respectively. The corresponding precision rates are 19.4%, 17.9%, and 16.4%. The success rate for catalytic sites is higher, with correct or partially correct predictions for 77.5%, 85.8%, and 88.2% of the enzymes in the test set, corresponding to the same respective Z score cut offs, if only the CatRes annotations are used as the reference set. Incorporation of additional literature annotations into the reference set gives total success rates of 89.9%, 92.9%, and 94.1%, again for corresponding cut-off values of 1.00, 0.99, and 0.98. False positive rates for a 75-protein test set are 1.95%, 2.60%, and 3.12% for Z score cut-offs of 1.00, 0.99, and 0.98, respectively. CONCLUSION: With a preferred cut-off value of 0.99, THEMATICS achieves a high success rate of interaction site prediction, about 86% correct or partially correct using CatRes/CSA annotations only and about 93% with an expanded reference set. Success rates for catalytic residue prediction are similar to those of other structure based methods, but with substantially better precision and lower false positive rates. THEMATICS performs well across the spectrum of E.C. classes. The method requires only the structure of the query protein as input. THEMATICS predictions may be obtained via the web from structures in PDB format at: http://pfweb.chem.neu.edu/thematics/submit.html. PMID- 17419879 TI - The lipid lowering effect of plant sterol ester capsules in hypercholesterolemic subjects. AB - BACKGROUND: Foods enriched with phytosterols have been proven to be an effective therapy to improve blood lipid profiles. However, none of the studies have investigated the efficacy in lipid lowering of plant sterol esters (PSE) in capsule form. The objective of this study is to determine if the plant sterol esters (PSE) in capsule form (1.3 grams of PSE/day) lowered plasma cholesterol levels and lipid ratios in free-living hypercholesterolemic subjects during a 4 week intervention period. METHODS: Sixteen subjects participated in a double blind, placebo-controlled, sequential study with a 4-week placebo phase followed by a 2-week wash-out period and a 4-week treatment phase. Subjects were instructed to maintain stable diet pattern and physical activities. Blood samples were collected at 7, 21 and 28 days of each phase. The primary measurements were change in plasma total cholesterol (TC), HDL-cholesterol (HDL) and LDL cholesterol (LDL) between phases and within each phase. The secondary measurements were change in triglycerides, lipoprotein ratios (TC/HDL, LDL/HDL) and C-reactive protein (CRP). RESULTS: In comparison to placebo, LDL-cholesterol was significantly reduced by 7% and 4% (P < 0.05) at both week 3 and week 4; HDL at week 3 of the treatment was significantly increased by 9% (P < 0.01), but not at week 4 (4%); total cholesterol was not significantly different from placebo throughout the period, TC/HDL and LDL/HDL were significantly reduced by (8%, 8%, 6%, 10%, respectively) (P < 0.01) at both week 3 and week 4. CRP and triglycerides did not differ either between the two phases or during the treatment phase. CONCLUSION: In conclusion, plant sterol ester capsule is effective in improving lipid profiles among hypercholesterolemic subjects in a free-living setting at the minimum dosage recommended by FDA. The significant improved lipid profiles were reached after three weeks of administration. To achieve better lipid lowering results, higher dosages and combination with diets low in saturated fat and cholesterol are recommended. PMID- 17419880 TI - Ethical challenges related to elder care. High level decision-makers' experiences. AB - BACKGROUND: Few empirical studies have been found that explore ethical challenges among persons in high public positions that are responsible for elder care. The aim of this paper was to illuminate the meaning of being in ethically difficult situations related to elder care as experienced by high level decision-makers. METHODS: A phenomenological-hermeneutic method was used to analyse the eighteen interviews conducted with political and civil servant high level decision-makers at the municipality and county council level from two counties in Sweden. The participants worked at a planning and control as well as executive level and had both budget and quality of elder care responsibilities. RESULTS: Both ethical dilemmas and the meaning of being in ethically difficult situations related to elder care were revealed. No differences were seen between the politicians and the civil servants. The ethical dilemmas mostly concerned dealings with extensive care needs and working with a limited budget. The dilemmas were associated with a lack of good care and a lack of agreement concerning care such as vulnerable patients in inappropriate care settings, weaknesses in medical support, dissimilar focuses between the caring systems, justness in the distribution of care and deficient information. Being in ethically difficult situations was challenging. Associated with them were experiences of being exposed, having to be strategic and living with feelings such as aloneness and loneliness, uncertainty, lack of confirmation, the risk of being threatened or becoming a scapegoat and difficult decision avoidance. CONCLUSION: Our paper provides further insight into the ethical dilemmas and ethical challenges met by high level decision-makers', which is important since the overall responsibility for elder care that is also ethically defensible rests with them. They have power and their decisions affect many stakeholders in elder care. Our results can be used to stimulate discussions between high level decision-makers and health care professionals concerning ways of dealing with ethical issues and the necessity of structures that facilitate dealing with them. Even if the high level decision-makers have learned to live with the ethical challenges that confronted them, it was obvious that they were not free from feelings of uncertainty, frustration and loneliness. Vulnerability was revealed regarding themselves and others. Their feelings of failure indicated that they felt something was at stake for the older adults in elder care and for themselves as well, in that there was the risk that important needs would go unmet. PMID- 17419881 TI - Clinical review: update of avian influenza A infections in humans. AB - Influenza A viruses have a wide host range for infection, from wild waterfowl to poultry to humans. Recently, the cross-species transmission of avian influenza A, particularly subtype H5N1, has highlighted the importance of the non-human subtypes and their incidence in the human population has increased over the past decade. During cross-species transmission, human disease can range from the asymptomatic to mild conjunctivitis to fulminant pneumonia and death. With these cases, however, the risk for genetic change and development of a novel virus increases, heightening the need for public health and hospital measures. This review discusses the epidemiology, host range, human disease, outcome, treatment, and prevention of cross-transmission of avian influenza A into humans. PMID- 17419882 TI - Clinical review: long-term noninvasive ventilation. AB - Noninvasive positive ventilation has undergone a remarkable evolution over the past decades and is assuming an important role in the management of both acute and chronic respiratory failure. Long-term ventilatory support should be considered a standard of care to treat selected patients following an intensive care unit (ICU) stay. In this setting, appropriate use of noninvasive ventilation can be expected to improve patient outcomes, reduce ICU admission, enhance patient comfort, and increase the efficiency of health care resource utilization. Current literature indicates that noninvasive ventilation improves and stabilizes the clinical course of many patients with chronic ventilatory failure. Noninvasive ventilation also permits long-term mechanical ventilation to be an acceptable option for patients who otherwise would not have been treated if tracheostomy were the only alternative. Nevertheless, these results appear to be better in patients with neuromuscular/-parietal disorders than in chronic obstructive pulmonary disease. This clinical review will address the use of noninvasive ventilation (not including continuous positive airway pressure) mainly in diseases responsible for chronic hypoventilation (that is, restrictive disorders, including neuromuscular disease and lung disease) and incidentally in others such as obstructive sleep apnea or problems of central drive. PMID- 17419883 TI - Counting repetitions: an observational study of outpatient therapy for people with hemiparesis post-stroke. AB - Given the contemporary clinical belief that more practice is better, it is important to determine how much practice currently occurs during physical therapy (PT) and occupational therapy (OT). The purpose of this study was to examine the number of repetitions of various activities during PT and OT outpatient treatment sessions for people with hemiparesis post-stroke. We observed 36 treatment sessions and recorded the types of activities and the number of repetitions of each activity that were done. Observations were categorized and descriptive statistics were generated for each category and subcategory. Our results showed that treatment time averaged 36 minutes per session. In sessions addressing the upper extremity, the average number of repetitions per session were 39 for active exercise movements, 34 for passive-exercise movements, and 12 for purposeful movements. In sessions addressing the lower extremity, the average number of repetitions per session were 33 for active-exercise movements, six for passive exercise movements, and eight for purposeful movements. In sessions addressing gait, the average number of steps taken was 292. In sessions addressing transfers, the average number of repetitions per session was 11. For most categories, there was considerable variability in the number of repetitions observed. We conclude that the numbers of repetitions observed during PT and OT for people with hemiparesis post-stroke are relatively small, except for gait steps. The fact that the number of repetitions of upper extremity purposeful movements was smaller than the number of repetitions of upper extremity active- and passive-exercise movements was surprising. This finding is inconsistent with current teaching that practice of purposeful movements is an integral part of improving functional status. PMID- 17419884 TI - Constraints on practice. PMID- 17419885 TI - Weight-bearing exercise accuracy influences muscle activation strategies of the knee. AB - PURPOSE: Dynamic stability of the knee joint is a research topicof increasing focus after ACL injury, stroke, and incomplete spinal cord injury. Since rehabilitation programs use functional weightbearing tasks to improve neuromuscular control of the knee, it is important to understand the adaptability of muscle control strategies during weight-bearing exercise. The purpose of this study was to compare muscle activation patterns during a single leg squat (SLS) exercise performed before and after feedback-controlled training. METHODS: This was a cross-sectional comparative study. Fifteen young, healthy individuals performed the SLS exercise while tracking a sinusoidal target with flexion and extension of the knee. The SLS instrument provided bidirectional resistance that was normalized to body weight. Six trials of 10 repetitions of the SLSs were performed to quantify improved performance (learning). Electromyographic activity from five muscles that cross the knee was analyzed. Accuracy of performance was measured by calculating the error between the target and actual knee displacement. RESULTS: Reduction in error measurements verified that individuals increased the accuracy of performance in each trial and retained this improvement across trials (p < 0.05). Modulation in muscle activity as a result of learning was reflected mainly in the biceps femoris, rectus femoris, and vastus lateralis muscles. CONCLUSION: Increased accuracy with the SLS exercise was accompanied by a decrease in coactivation of selected musculature around the knee. This study presents a novel approach to quantify the effect of performance on muscle synergistic activation patterns during weight-bearing exercise. Controlled strengthening, as defined in this study, emphasizes accuracy of performance in conjunction with principles of strength training and has implications to knee control. PMID- 17419886 TI - The Kinesthetic and Visual Imagery Questionnaire (KVIQ) for assessing motor imagery in persons with physical disabilities: a reliability and construct validity study. AB - PURPOSE: To benefit from mental practice training after stroke, one must be able to engage in motor imagery, and thus reliable motor imagery assessment tools tailored to persons with sensorimotor impairments are needed. The aims of this study were to (1) examine the test-retest reliability of the Kinesthetic and Visual Imagery Questionnaire (KVIQ-20) and its short version (the KVIQ-10) in healthy subjects and subjects with stroke, (2) investigate the internal consistency of both KVIQ versions, and (3) explore the factorial structure of the two KVIQ versions. METHODS: The KVIQ assesses on a five-point ordinal scale the clarity of the image (visual: V subscale) and the intensity of the sensations (kinesthetic: K subscale) that the subjects are able to imagine from the first person perspective. Nineteen persons who had sustained a stroke (CVA group) and 46 healthy persons (CTL group) including an age-matched (aCTL: n = 19) control group were assessed twice by the same examiner 10 to 14 days apart. The test retest reliability was assessed using intraclass correlation coef- ficients (ICCs). The internal consistency (Cronbach alpha) and the factorial structure of both KVIQ versions were studied in a sample of 131 subjects. RESULTS: In the CVA group, the ICCs ranged from 0.81 to 0.90, from 0.73 to 0.86 in the aCTL group, and from 0.72 to 0.81 in the CTL group. When imagining movements of the affected and unaffected limbs (upper and lower limbs combined) ICCs in the CVA group ranged, respectively, from 0.71 to.87 and from 0.86 to 0.94. Likewise, when imagining movement of the dominant and nondominant limbs, ICCs in the aCTL group ranged, respectively, from 0.75 to 0.89 and from 0.81 to.92. Cronbach a values were, respectively, 0.94 (V) and 0.92 (K) for the KVIQ-20 and 0.89 (V) and 0.87(K) for the KVIQ-10. The factorial analyses indicated that two factors explained 63.4% and 67.7% of total variance, respectively. CONCLUSION: Both versions of the KVIQ present similar psychometric properties that support their use in healthy individuals and in persons post-stroke. Because the KVIQ-10 can be administered in half the time, however, it is a good choice when assessing persons with physical disabilities. PMID- 17419887 TI - Physical therapy management for conversion disorder: case series. AB - PURPOSE: Patients with conversion disorder present with symptoms of sensory and motor dysfunction that are not explained by known physical disorders or pathophysiological mechanisms. Correct diagnosis and prompt, effective management of patients with this disorder is essential. Healthcare providers may be unaware of the important role of physical therapy in the management of patients with conversion disorder. While numerous reports have suggested the need for physical therapy management of this disorder, there is a lack of reports outlining specific physical therapy management principles or daily treatment progression. The purpose of this series of case reports is to provide an overview of conversion disorder and describe successful physical therapy management strategies used to treat three patients with movement impairment due to this disorder. CASE DESCRIPTION: The patients were 18-, 20-, and 34-year-old women treated in a hospital inpatient rehabilitation setting. Behavioral modification and shaping techniques formed the basis for the physical therapy treatment approach. Abnormal movement patterns were ignored, and correct movement patterns were reinforced using feedback and praise. The patients advanced through a progressively more difficult therapy program based on treatment approaches used with analogous neurological conditions. OUTCOMES: All three patients showed complete resolution of their symptoms and returned to independent mobility, independent living, and to work or school. PMID- 17419888 TI - Exploring barriers to remaining physically active: a case report of a person with multiple sclerosis. AB - Physical therapy intervention for those with chronic disabling conditions typically follows an episode of care approach: therapists provide services when a decrement in functional performance occurs such that individuals require intervention to return to baseline performance. Attention to the psychosocial supports required for successful transition can be unintentionally minimized when the focus of an episode of care follows a change in physical function. The purpose of this case report is to present and discuss the challenges to successful community reintegration following physical therapy intervention with an emphasis on developing independent exercise habits in management of a person with multiple sclerosis. RW, presented in this case study, is a 52-year-old man diagnosed with progressive multiple sclerosis five years before self-referral to a pro bono physical therapy clinic. PMID- 17419889 TI - Effect of supplementation of women in high-risk pregnancies with long-chain polyunsaturated fatty acids on pregnancy outcomes and growth measures at birth: a meta-analysis of randomized controlled trials. AB - It is hypothesized that the intake of long-chain PUFA (LC-PUFA) throughout pregnancy could protect against pre-eclampsia, pregnancy-induced hypertension or intra-uterine growth retardation, and is essential for optimal neural development. The objective of the present study was to systematically evaluate the effect of LC-PUFA supplementation of high-risk pregnant women's diets on pregnancy outcomes and growth measures at birth. We searched MEDLINE, EMBASE, CINAHL and the Cochrane Library through March 2006 and references in reviewed articles for randomized controlled trials (RCT) comparing LC-PUFA supplementation with placebo or no supplementation in women with high-risk pregnancies. We found no evidence that supplementation influenced the duration of pregnancy or the percentage of preterm deliveries < 37 weeks of gestation. However, compared with controls, supplementation was associated with a significantly lower rate of early preterm delivery ( < 34 weeks of gestation) (two RCT; n 291; relative risk 0.39 (95 % CI 0.18, 0.84)). There was no significant difference in the infant birth weight, the rate of low birth weight ( < 2500 g or < 10th percentile) and the recurrence of intra-uterine growth retardation. Other pregnancy outcomes (for example, the rate of pregnancy-induced hypertension, the rate of pre-eclampsia and the rate of Caesarean section) were also similar in both groups. In conclusion, the present data suggest that supplementation with n-3 LC-PUFA in women with high-risk pregnancies reduced the risk of early preterm delivery in the fatty acid-supplemented group compared with the placebo group, while no other effects on pregnancy outcomes were detected. PMID- 17419890 TI - Estimation of human daily boron exposure in a boron-rich area. AB - Although, the safe limits of human daily boron (B) exposure are not absolutely clear, there is a growing interest in B and its effects on human health. The aim of the present study was to estimate daily B exposure in 66 males in Turkey living in a B-rich area using water containing at least 2 mg/l boron, with an average age of 38.55 (se 1.66) years and an average number of years of residence in the B-rich area of 35.89 (se 1.73). Another group of males (n 57), living in the city centres of Balikesir and Ankara, were taken as controls; the average age and number of years of residence for this group were 29.44 (se 1.43) and 10.26 (se 1.83) years, respectively. As it is assumed that the B level in urine reflects daily B exposure, the amount of urinary B of both the study and control groups was analysed by using an inductively coupled plasma optical emission spectrometry (ICP-OES) technique. The average daily B exposure value was calculated as 6.77 (se 0.47) mg in the study group and 1.26 (se 0.1) mg in the controls. The results of this study are expected to contribute to creating a reference value for a safe daily B exposure. PMID- 17419891 TI - Supranormal dietary folic acid supplementation: effects on methionine metabolism in weanling rats. AB - There are nationwide folic acid (FA) fortification programmes of staple foods established or under consideration in order to prevent neural tube defects. Universal FA fortification still remains controversial because of the concern that additional FA in the diets of population groups (e.g. children and elderly) not initially targeted for fortification may suffer adverse effects. However, dietary surveys regarding folate generally deal with adults and little is known about the consumption and long-term effects of fortified food and supplements in growing individuals. Recent reports from our laboratory show several effects of high-dose folate supplementation in rats. In the present work, we studied the effect of FA on the methionine cycle in weanling (3-week-old) male rats after 4 weeks of supplementation with 40 mg FA/kg diet v. control (1 mg FA/kg diet). FA supplementation resulted in a reduction of homocysteine and creatinine concentrations v. control group. FA supplementation did not alter S adenosylmethionine/S-adenosylhomocysteine ratio, DNA methylation, enzymatic activities or concentrations of vitamins involved in the nutritional regulation of the methionine cycle, except for folate. FA supplementation of 40 mg/kg did not lead to hepatic or renal damage. In conclusion, there were no apparent adverse effects on one-carbon metabolism after FA supplementation in the studied conditions. PMID- 17419892 TI - Consumption of fish and risk of colon cancer in the Norwegian Women and Cancer (NOWAC) study. AB - Recent studies have shown a decreased risk of colon cancer with consumption of fish. However, most studies on fish consumption do not distinguish between lean and fatty fish, or between poached and fried fish. The aim of this study was to investigate any association between fish consumption and colon cancer in The Norwegian Women and Cancer (NOWAC) study. We focused mainly on lean fish, which was further divided into poached and fried fish. A total of 63 914 women were included in the analysis, 254 of whom were found to have colon cancer during follow-up. Since fish consumption was one of the main topics of interest in the NOWAC study, there is a predominance of women from northern Norway due to higher fish intake in this area; hence the study is not representative of the whole of Norway. The participants completed a semi-quantitative FFQ between 1996 and 1999, and were followed-up for incidence of colon cancer until 2004. No association between fish consumption and risk of colon cancer was seen, except for the third tertile of poached lean fish consumption (relative risk (RR) 1.46, 95 % CI 1.04, 2.06). This association disappeared when excluding women with less than 1 year of follow-up. In conclusion, the present study does not support the hypothesis of a protective effect of fish against colon cancer risk. PMID- 17419893 TI - Choosing a paediatric tracheostomy tube: an update on current practice. AB - OBJECTIVES: A variety of paediatric tracheostomy tubes are available. This article reviews those in current use at Great Ormond Street Hospital. METHODS: We outline our preferences and the particular indications for the different tubes, speaking valves and other attachments. RESULTS: Practice has changed significantly in recent years. One product has been re-sized by its manufacturer; others are no longer commonly used. An updated sizing chart is included for reference purposes, together with manufacturers' contact details. CONCLUSIONS: The choice of paediatric tracheostomy tube is driven by clinical requirements. A small range of tubes are suitable for the majority of children, but some will require other varieties in specific circumstances. PMID- 17419894 TI - Unusual presentation of temporal bone involvement in Churg-Strauss syndrome. AB - OBJECTIVE: To present a case of a 60-year-old male with a history of sudden onset sensorineural hearing loss due to Churg-Strauss syndrome. CASE REPORT: The patient had a 20-year history of asthma and recurrent right otitis media and a nasal polypectomy four years prior to presenting with ear symptoms. Ear, nose and throat involvement is common in Churg-Strauss syndrome, usually manifesting as allergic rhinitis and chronic rhinosinusitis with or without polyps. CONCLUSIONS: Otolaryngologists play an important role in making an early diagnosis of this disease. To our knowledge this is the first case of Churg-Strauss syndrome primarily presenting with otological pathology: left sensorineural hearing loss and right otitis media. PMID- 17419895 TI - Nasopharyngeal carcinoma presenting as carotidynia in an 18-year-old patient. AB - INTRODUCTION: Nasopharyngeal carcinoma can present with a variety of symptoms the most common being painless cervical lymph node metastases, epistaxis and conductive hearing loss. CASE REPORT: We report a case of an 18-year-old male patient presenting to the ENT department with left sided carotidynia - severe pain in the neck radiating up to his ear. He had a small, exquisitely tender lump just below the bifurcation of the left common carotid artery. After the appropriate investigations he was diagnosed with nasopharyngeal carcinoma. DISCUSSION: In a literature search on Medline and Pubmed we found no reported cases of nasopharyngeal carcinoma presenting with carotidynia, nor was carotidynia ever found to be associated with nasopharyngeal carcinoma. CONCLUSION: We believe that this is the first reported case of such a presentation of nasopharyngeal carcinoma. PMID- 17419896 TI - Sudden sensorineural hearing loss in a patient with primary antiphospholipid syndrome. AB - OBJECTIVE: Despite multiple systemic manifestations, sudden sensorineural hearing loss in a patient with antiphospholipid syndrome is rarely reported. PATIENT: A 46-year-old man with primary antiphospholipid syndrome had a sudden onset of hearing loss and tinnitus in the right ear in December 2005, because he discontinued use of warfarin and acetylsalicylic acid for a few days. RESULTS: Audiometry revealed saucer-type sensorineural hearing loss with a pure tone average of 73 dB in the right ear, and flat-type hearing loss with a pure tone average of 25 dB in the left ear. Electronystagmography displayed multiple central signs and bilateral canal paresis, while a vestibular evoked myogenic potential test revealed bilateral delayed responses. After admission, the patient was re-treated with warfarin and acetylsalicylic acid. Follow-up audiometry showed recovery of right-sided hearing, with a pure tone average of 12 dB, three days after presentation. CONCLUSION: Consensus exists on the effectiveness of anticoagulant agents in aiding a favourable outcome of sudden sensorineural hearing loss in patients with antiphospholipid syndrome. PMID- 17419897 TI - Intranasal myopericytoma. A tumour with perivascular myoid differentiation: the changing nomenclature for haemangiopericytoma. AB - We present a case report of a patient who developed a sinonasal myopericytoma treated by surgical excision through a lateral rhinotomy. Some aggressive features on pre-operative computed tomography scanning and the complexity of recent changes in the histological nomenclature for these tumours led to consideration of adjuvant therapy. The close histological relationship between myopericytoma, myofibromatosis, solitary myofibroma and infantile haemangiopericytoma is discussed. This group of lesions constitute a single morphological spectrum with differentiation towards perivascular myoid cells (pericytes). Currently myopericytoma is the most appropriate and accepted term embracing all these entities. A review of the literature has been reassuring in identifying these tumours as benign but with a reasonably high rate of local recurrence (17 per cent). The treatment of choice is surgical excision with further excisions for local recurrence. PMID- 17419898 TI - Role of otoacoustic emission in children with middle-ear effusion and grommets. AB - OBJECTIVE: To evaluate the role of otoacoustic emission in children with middle ear effusion and grommets. MATERIALS AND METHODS: A prospective study was carried out on a total of 90 ears. All children listed for grommet insertion had a pre operative and post-operative (three to six months after grommet insertion) pure tone audiometry, tympanometry and otoacoustic emission recorded. A comparison was made between pure tone audiometry and otoacoustic emission both pre-operatively and post-operatively. RESULTS: Pre-operatively, 63 ears had an abnormal pure tone audiometry of which 59 had absent otoacoustic emission. Therefore the sensitivity of otoacoustic emission in detecting a conductive loss was 59/63 = 94 per cent (95 per cent confidence interval 85 to 98 per cent). All 27 ears with normal hearing pre-operatively had normal otoacoustic emission. The specificity of otoacoustic emission was 27/27 = 100 per cent, (95 per cent confidence interval, 88 to 100 per cent). The positive predictive value was 59/59 = 100 per cent (95 per cent confidence interval, 94 to 100 per cent). After three to six months all post-operative patients with grommets had a normal pure tone audiometry and otoacoustic emission. So both pure tone audiometry and otoacoustic emission were strongly related both in patients with middle-ear effusion and in patients with grommets. CONCLUSION: As the demonstration of hearing in young and difficult-to test children can be problematic and time-consuming, we suggest that otoacoustic emission can be used as an alternative to pure tone audiometry in patients with middle-ear effusion and grommets. PMID- 17419899 TI - Microdebrider removal of tracheal papilloma via tracheostomy in the child with an obliterated larynx. AB - In children, recurrent respiratory papillomatosis is the most common benign neoplasm of the airway. The disease frequently involves the larynx and may spread to extralaryngeal sites. Use of a microdebrider has been suggested as a safe and low-cost technique which reduces operating time, compared with laser removal of laryngeal lesions. We describe a technique for using a microdebrider to remove tracheal papillomas when the larynx is obliterated with the disease. PMID- 17419900 TI - Correlation between a foreign body in the external auditory canal and otitis media with effusion. AB - OBJECTIVE: To determine if there is a relationship between a foreign body in the external auditory canal and undiagnosed otitis media with effusion or significant eustachian tube dysfunction in children. STUDY DESIGN AND SETTING: This is a prospective, uncontrolled analysis of 37 consecutive children with a foreign body in the external auditory canal (group I) and 37 children with non-ENT complaints as a control (group II), seen over two years in the ENT unit of a district general hospital. All the children underwent removal of the foreign body, examination of both ears by a senior ENT surgeon followed by tympanometry within seven to 10 days. RESULT: Thirty-seven children with a foreign body in the external auditory canal (group I) were analysed and a similar number of children with non-ENT problems (group II) were taken as a control. The age range for both groups was two years to 10 years with a median age of six years. Of the 37 children, 25 (68 per cent) in group I had an abnormal view of the tympanic membrane compared to only five (14 per cent) in group II (p < 0.04-chi-squared test). In group I 20 children (54 per cent) and in group II three children (8 per cent) had abnormal middle-ear compliance (either type B or type C2) (p < 0.05-chi squared test). Nineteen (51 per cent) children in group I had a history of previous ear symptoms such as irritation, otalgia, blockage or deafness more than once in the past six months, and none had in the control group (p < 0.03-chi squared test). CONCLUSION: There is clinical and statistical evidence to suggest that children may insert a foreign body in the ear as a result of irritation/pressure sensation secondary to otitis media with effusion or significant eustachian tube dysfunction. Therefore, we recommend that all children with a history of a foreign body in the ear should be screened in an ENT clinic. PMID- 17419901 TI - [Errors and complications in thyroid surgery: analogies and differences]. PMID- 17419902 TI - T-level downstaging and complete pathologic response after preoperative long-term radiochemotherapy for locally advanced rectal cancer. AB - Advantages of neoadjuvant chemoradiotherapy for locally advanced carcinoma of the middle and the lower third of the rectum are downstaging and downsizing of the tumor. Results of pathologic results are affected by post-treatment tissue changes and may influence the choice of surgical procedure. Forty-three consecutive patients (27 male, 16 female; mean age 64 years) were operated after receiving a long-term chemoradiotherapy during a period of 16 months. The data of initial staging procedure (high resolution magnetic resonance imaging) and results of pathological examination of the surgical specimens were analyzed. Regression of tumor was assessed by the absence of vital tumor cells and the post treatment fibrotic tissue alterations. Regression of tumor size was seen in 42/43 patients leading to an improved T-stage in 27 patients. R0-resection was possible in all cases, although there was a perirectal tumor infiltration to less than 2 mm to circumference of the surgical specimen in 2 cases and unexpected small liver metastasis in 5 cases. Complete remission rate was 23.3% (10 cases). Detecting small amounts of vital tumor cells in altered tissue after chemoradiotherapy is a major problem of pathological examination procedure and should be taken into consideration by the surgeons. The choice of operation (resection vs. abdominoperineal extirpation vs. local excision) should be committed to the initial imaging procedure and not to any restaging procedure after neoadjuvant chemoradiotherapy. PMID- 17419903 TI - [Adenocarcinoma of the appendix. A case report and review of the literature]. AB - Primary adenocarcinoma of the appendix is a rare malignancy that constitutes less than 0.5% of all gastrointestinal neoplasms. Usually the diagnosis is made only after histological examination of surgically removed inflamed appendix. Alternatively represent an unexpected finding, confirmed by frozen section, during surgery performed for acute appendicitis or other non appendiceal pathologies. Natural history is strongly influenced by anatomic peculiarities of the appendix that predispose to early spread and perforation. Frequently is associated with synchronous and metachronous colorectal or extraintestinal cancers. The correct management is the right hemicolectomy as a primary procedure in the case of preoperatively or intraoperatively diagnosis or as secondary procedure, after two-three weeks from appendectomy, when the microscopic examination of specimen reveals the presence of adenocarcinoma. Right hemicolectomy is the best treatment for all histologic types (colonic, mucinous, adenocarcinoid), in presence of perforation and even in Dukes A tumors. A careful intraoperative search for synchronous lesions and a life-long program of surveillance for the detection of early stage metachronous carcinomas are recommended. The Authors report a case of primary adenocarcinoma of the appendix occurred in a 78 year-old female patient, diagnosed incidentally during surgery performed for ileus from suspected cecal neoplasm. PMID- 17419904 TI - [Is extrauterine endometriosis confined to the gynecological sphere? A critical review of the experience in a general surgery unit]. AB - Extrauterine or external endometriosis (e.e.) describes ectopic localization of functional endometrial tissue, a finding whose incidence is increasing due to the diffusion of laparoscopic procedures. The clinical presentation of such disease is often non-specific, even in those cases with a definite surgical indication, depending on the site and pathology of the lesions. Surgical planning is therefore difficult at times, specifically regarding the extent of resection in patients--as young women--willing to maintain fertility. The Authors report on 7 cases observed in their own experience (inguinal endometriosis--1 case, umbilical endometriosis--1 case, abdominal wall endometriosis--3 cases, intestinal endometriosis--1 case, diaphragmatic endometriosis associated with pneumothorax- 1 case), representing the wide range of clinical settings related to e.e. Based on literature data, an analysis of clinical and diagnostic issues is carried out. Specifically, the problems related to extent of surgical resection in multifocal cases, possible malignant degeneration and localization on abdominal wall scars are discussed. PMID- 17419905 TI - [Surgical treatment of sinus pilonidalis by Dufourmentel's flap technique]. AB - The present study reports the results of surgical treatment of 175 consecutive chronic pilonidalis sinus cases by excision alone, excision and primary closure, excision and Dufourmentel's flap technique. The analysis of hospital stay , average recovery time, patient compliance and percentages of failures and recurrences enables us to state that Dufourmentel's technique is the best choice, allowing fast recovery with minimal discomfort and lower rate of recurrence. Moreover the good compliance of the technique and the use of recently introduced more effective local anaesthetics have allowed us to plan the treatment in a day surgery setting with good clinical results. PMID- 17419906 TI - Surgical treatment with Ligasure Precise of schwannoma of brachial plexus: case report. AB - English version We present a case of bulky schwannoma arising from the brachial plexus treated by a new surgical device. A 38-year-old man presented with a slow growing left-sided supraclavicular mass and complained paresthesia of the third and forth fingers of the hand and forearm weakness. Physical examination revealed Tinel's sign. A CT-scan revealed a solid mass situated in the left profound supraclavicular fossa. The tumour was resected with the utilization of bipolar vessel sealing system (Ligasure Precise). This device is very useful in sutureless removal of masses localized in deep supraclavicular fossa. During the operation, care was taken to preserve the nerve function. PMID- 17419907 TI - [The ATLS Courses in Italy. Twelve years experience]. AB - ATLS Courses were introduced in the USA in 1980 and have been taught in Italy since 1994. Through theoretical lessons and practical sessions, their scope is to provide proper training for doctors with every kind of speciality who work in Emergency Departments, in order to prepare them to rapidly and effectively intervene on a patient who has suffered a serious trauma. Universities, in fact, do not prepare doctors adequately on this topic, while the application of the ATLS method in the first hours after trauma can effectively improve the prognosis of the patient. This study collects the data of the Italian experience in ATLS training, which has been carried out under the aegis of the Italian Chapter of the American College of Surgeons. The ATLS Courses have become widespread throughout our Country, which today is the fourth in the world for number of courses held every year. PMID- 17419908 TI - [Modern orientation in parotid surgery]. AB - The choice of the surgical approach is very relevant in parotid surgery because of the extreme anatomic variability of the parotideal district and the functional importance of the branches of facial nerve. The aim of this paper is to prove how an accurate knowledge of the anatomy and a careful execution of surgery permit radicality in case of tumors and respect of nerves and other structures of the region. We studied anatomy of the different intra- and periparotideal structures to compare two different approaches in parotidectomy, proving how the orthograde technique is safer and more effective in preventing hyatrogenous nerve injuries and other major complications. PMID- 17419909 TI - [Usefulness of endobronchial ultrasonography with real-time needle aspiration for lung cancer staging]. AB - BACKGROUND AND OBJECTIVE: To determine the usefulness of endobronchial ultrasonography (EBUS) with real-time needle aspiration (NA) for lung cancer staging. PATIENTS AND METHOD: All patients examined with EBUS and real-time NA to measure and sample mediastinal and lobar nodes for lung cancer staging during one year were included, independently of the size of the mediastinal nodes at computed tomography (CT). RESULTS: Eighty two nodes > 5 mm were sampled using EBUS-NA (16.0 [7.2] mm; 23 cases or= 2, according to the National Nosocomial Infections Surveillance (NNIS) (OR, 3.3; 95% confidence interval [CI], 1.1-10.8) and post-operative non-infectious complications (OR, 8.9; 95% CI, 1.1-83.1) (p < 0.05). The global infection rate was 2.4% (95% CI, 1.6%-3.2%) of 1,666 joint implants. Gram-positive cocci were the principal involved microorganisms (63%). CONCLUSIONS: A high NNIS together with non infectious post-operative complications increase the risk of prosthesis infection. PMID- 17419912 TI - [Prognostic value of high blood pressure in diabetic patients]. PMID- 17419913 TI - [New computerized languages for information on drugs]. PMID- 17419914 TI - [Social pharmacology of the information of prescription drugs on the internet]. PMID- 17419915 TI - [New therapeutic aspects for the control of risk factors in obesity]. AB - Treatment of obesity continues to rely upon the classical triad of nutritional advise, increase of physical activity and use of drugs. However, in recent years, there have appeared novelties in both therapeutic targets and new molecules. With regard to therapeutic targets of obesity, success does not consist of losing much weight but attaining a moderate yet maintained weight lose (5-10% of initial weight) at the expense of visceral fat. In other words, instead of weight, what is needed is a waist reduction, mainly to improve or prevent obesity-related metabolic and vascular complications. Regarding drugs, a new molecules is about to appear in the international pharmaceutical market: rimonabant. A selective blocker of the endocannabinoid receptor CB1, it has proven to be effective and safe in treating obesity and its comorbidities. Another known agent, orlistat, has proven to be effective in the prevention of the development of type 2 diabetes in obese patients with or without glucose intolerance. PMID- 17419916 TI - [Tetanus prophylaxis in an emergency medical service: a quality improvement proposal]. PMID- 17419917 TI - [Hemochromatosis: controversy for blood donation]. PMID- 17419918 TI - [Value of autopsy in heart transplant recipients]. PMID- 17419919 TI - [Suicide prevention in primary care: psychopharmacological treatment]. PMID- 17419920 TI - [Anal leiomiosarcoma associated to Lynch syndrome]. PMID- 17419921 TI - [Gabapentin-induced central hypoventilation]. PMID- 17419922 TI - [Bleomicine induced dermatosis flagellata]. PMID- 17419923 TI - [Evaluation in public health: does anything go?]. PMID- 17419924 TI - [Cost-effectiveness of the addition of acarbose to the treatment of patients with type-2 diabetes in Spain]. AB - OBJECTIVES: To assess the cost-effectiveness of the addition of acarbose to existing treatment in patients with type 2 diabetes mellitus (DM2) in Spain. METHODS: The CORE Diabetes Model (a published and validated computer simulation model) was used to project long-term clinical and cost outcomes in DM2. Transition probabilities and risk adjustments were derived from published sources. Treatment effects and baseline cohort characteristics were based on a meta-analysis. Direct costs were retrieved from published sources and projected over patient lifetimes from the perspective of the Spanish National Health Service. Costs and clinical benefits were discounted at 3% per year. Sensitivity analyses were performed. RESULTS: Acarbose treatment was associated with improved life expectancy (0.23 years) and quality-adjusted life years (QALY) (0.21 years). Direct costs were on average euro 468 per patient more expensive with acarbose than with placebo. The incremental cost-effectiveness ratios were euro 2,002 per life year gained and euro 2,199 per QALY gained. An acceptability curve showed that with a willingness to pay euro 20,000, which is generally accepted to represent very good value for money, acarbose treatment was associated with a 93.5% probability of being cost-effective. CONCLUSIONS: This long-term economic study showed that the addition of acarbose to existing therapy for DM2 was associated with improvements in life expectancy and QALYs in these patients. PMID- 17419926 TI - [Design and validation of a home care satisfaction questionnaire: SATISFAD]. AB - OBJECTIVES: To design and validate an instrument to assess satisfaction with home care services, in both self-administered and telephone versions. METHODS: We performed a cross-sectional observational study of the population using home care services in the health districts of Malaga, Costa del Sol, Almeria and Granada (Spain). A questionnaire was designed by an expert panel using a Deplhi technique. Reliability between the self-administered and telephone versions was analyzed. Finally, internal consistency and construct validity were assessed. RESULTS: Reliability between the self-administered and telephone versions was high (intraclass correlation coefficient = 0.876; 95% CI, 0.726-0.941; p = 0.0001). Internal consistency was adequate (Cronbach's alpha: 0.853 and 0.799 for both versions, with or without caregiver, respectively). The factorial analysis explained 66.80% and 67.81% of the observed variance for the two versions (with or without caregiver, respectively). Two factors were isolated and related to interpersonal relationships, the role of the carer, and decision making. CONCLUSION: Assessment of satisfaction with home care can be performed with the dimensions routinely used in satisfaction studies, but these should be evaluated with instruments designed ad hoc. Accessibility, communication and interpersonal relationships have a high explanatory value in satisfaction among this population. PMID- 17419927 TI - [Barriers and facilitators to health care coordination in two integrated health care organizations in Catalonia (Spain)]. AB - OBJECTIVES: To analyze 2 integrated delivery systems (IDS) in Catalonia and identify areas for future development to improve their effectiveness. METHODS: An exploratory, descriptive, qualitative study was carried out based on case studies by means of document analysis and semi-structured individual interviews. A criterion sample of cases and, for each case, of documents and informants was selected. Study cases consisted of the Consorci Sanitari del Maresme (CSdM) and the Consorci Sanitari de Terrassa/Fundacio Hospital Sant Llatzer (FHSLL). A total of 127 documents were analyzed and 29 informants were interviewed: IDS managers (n = 10), technical staff (n = 5), operational unit managers (n = 5) and health professionals (n = 9). Content analysis was conducted, with mixed generation of categories and segmentation by cases and subjects. RESULTS: CSdM and CSdT/FHSLL are health care organizations with backward vertical integration, total services production, and real (CSdM) and virtual (CSdT/FHSLL) ownership. Funds are allocated by care level. The governing body is centralized in CSdM and decentralized in CSdT/FHSLL. In both organizations, the global objectives are oriented toward improving coordination and efficiency but are not in line with those of the operational units. Both organizations present a functional structure with integration of support functions and utilize mechanisms for collaboration between care levels based on work processes standardization. CONCLUSIONS: Both IDS present facilitators and barriers to health care coordination. To improve coordination, changes in external elements (payment mechanism) and in internal elements (governing body role, organizational structure and coordination mechanisms) are required. PMID- 17419928 TI - [Distribution of the public healthcare budget of Catalonia (Spain) for 2005 among the 17 ICD-9-CM categories]. AB - OBJECTIVES: To describe the distribution of the Catalan public healthcare budget for 2005 among the 17 ICD-9-CM (International Classification of Diseases, Ninth revision, clinical modification) categories. MATERIAL AND METHOD: The methodology comprised 2 phases: an initial phase in which the global budget was distributed by type of healthcare (hospital, outpatient or pharmacological care), and a second phase in which the expenditure was distributed by the type of care among the ICD-9-CM categories. In the first phase, this distribution was based on information enabling the various budget items to be assigned to the different types of care. Various elements were used for the distribution by categories, depending on each type of care: hospital stay, outpatient visit or consumption by therapeutic subgroup. RESULTS: Distribution of the budget was as follows: 46.6% for specialized care, 27.5% for pharmacological care, and 20.0% for primary care; 5.9% was not distributed. Of the 17 categories, that accounting for the largest percentage (17.3%) was "diseases of the circulatory system" (VII), followed by category VIII, "diseases of the respiratory system" which totaled 10.9%. The budget was concentrated in 5 categories, the 2 mentioned above plus category V "mental disorders" (9.4%), category II "tumors" (9.1%) and category IX "disorders of the digestive system" (7.7%), which accounted for 54.4% of the total budget. The internal composition of each category showed major variations. CONCLUSION: The distribution of the budget offers a point of reference for health planning and management. PMID- 17419929 TI - Influence of available resources on medical practitioners' decision-making process and practice: study of a reference hospital emergency department. AB - OBJECTIVE: To evaluate variations in the observation period in the emergency department (ED) in response to bed availability. METHODS: A quasi-experimental pre-test post-test study without a control group was conducted in the ED observation ward over 2 1-month periods. During this time the only variable that changed was the number of beds available, which decreased from 20 (pre-test period) to 16 (post-test period). RESULTS: The ED attended 7,725 patients: 3,706 patients in pre-test period, 335 of whom were admitted to the observation ward, with an average length of stay of 1,105.4 minutes per patient, and 4,019 patients in post-test period, 570 of whom were admitted to the observation ward, with an average length of stay of 686.1 minutes per patient (p < 0.001). There was no variation in mortality, re-admissions or complaints. CONCLUSIONS: A reduction in bed availability for observation purposes shortens patient length of stay. PMID- 17419930 TI - [An extended version of the Font-Roja job satisfaction questionnaire]. AB - INTRODUCTION: Several studies have shown the need to include the physical work environment among the dimensions included in job satisfaction evaluation. However, this dimension was not included in the Font-Roja questionnaire. The present study introduces two items exploring this dimension and adheres to the hypothesis that physical work environment has a significant impact on job satisfaction evaluation. METHOD: A total of 227 geriatric workers participated in this study. The participants completed the Font-Roja job satisfaction questionnaire with 2 additional items exploring the physical work environment. Factor analysis and principal components analysis with rotation varimax were used to determine the diverse components of job satisfaction. To determine the coherence of the scales and the consistency of the added items, Cronbach's a was used. These methods were applied to both questionnaires, the classical 24-item questionnaire and the extended 26-item questionnaire. RESULTS: The classical Font Roja questionnaire was composed of 8 factors, explaining 60.02% of the variance. The extended questionnaire was structured into 9 factors, explaining 61.81% of the variance. The new factor was composed of both added items. The internal consistency of the Font-Roja classical scale was alpha = 0.773 and that of the extended scale was alpha = 0.791. DISCUSSION: The extended scale is superior to the classical scale. The results obtained seem to support the hypothesis that, for analysis of job satisfaction, the instruments used should contain items on the physical work environment. PMID- 17419931 TI - [Sociodemographic characteristics of the immigrant population with a diagnosis of tuberculosis in the autonomous community of Madrid (Spain)]. AB - OBJECTIVE: To describe the social and demographic characteristics of immigrant patients diagnosed of tuberculosis in the Community of Madrid. PARTICIPANTS AND METHOD: Descriptive study. The information was gathered from the patients diagnosed in 15 hospitals of the Community of Madrid, and 3 primary health Red Cross facilities. Criteria for inclusion: to be tuberculosis diagnosed during the year 2003, to be economic immigrant, to be older than 15 year-old, and not to be in prison. RESULTS: 312 persons were identified. 296 were included in the study. Main nationalities: Ecuador (27.4%) and Morocco (19.3%). The 18.9% of the population does not have health insurance. The time between the arrival to Spain and the illness was 29.5 months. The 66.9% of the cases lived with some relatives. The 63.2% of the active population worked in the informal economy. The main risk factors was to be risk drinker. CONCLUSIONS: To know the social and sanitary situation of immigrant patients diagnosed of tuberculosis is base to adapt the control programs to a new epidemiologist and social situation. PMID- 17419932 TI - [Gaceta Sanitaria: a memory exercice]. PMID- 17419933 TI - [Health promotion: a profitable investment? Economic efficiency of preventive interventions in Spain]. AB - OBJECTIVE: To examine the quantity and quality of economic evaluations analyzing preventive interventions in Spain to September 2005, with the further goal of extracting conclusions for further research and the design of future programs. METHODS: We performed a systematic review of the evidence. First, we defined the criteria for including studies in our review. Second, a search was conducted of specialized search engines (Pubmed, NHS EED, DARE, HTA, HRSPROJ, IME, EMBASE) and a manual search was performed of journals and the web sites of Spanish public health organizations. In a third phase, the characteristics relevant to our analysis were extracted from the selected articles. Lastly, the characteristics collected were analyzed through uni- and bivariate analyses. RESULTS: Forty-nine articles were found that complied with the inclusion criteria, of which 40 were reviewed. The technique most extensively used was cost-effectiveness analysis (60% of all articles). Twenty-eight evaluations (70%) focused on immunization campaigns. The quality of publications increased overtime, from an average score of 4.21 (1985-1995) to 6.38 (1995-2004), although several methodological areas still require improvement. Lastly, 72.5% of the studies supported the universal use or expansion of the policy analyzed. CONCLUSIONS: The Spanish research community should increase efforts to improve the quantity and quality of economic evaluations in preventive health. Three basic strategies are suggested: a) evaluation of the preventive programs currently in place in Spain and dissemination of the results; b) efforts to publish and index articles in international scientific journals; and c) adherence to international economic evaluation guidelines and manuals. PMID- 17419934 TI - [Air pollutant exposure during pregnancy and fetal and early childhood development. Research protocol of the INMA (Childhood and Environment Project)]. AB - INTRODUCTION: The INMA (INfancia y Medio Ambiente [Spanish for Environment and Childhood]) project is a cooperative research network. This project aims to study the effects of environment and diet on fetal and early childhood development. This article aims to present the air pollutant exposure protocol during pregnancy and fetal and early childhood development of the INMA project. METHODS: The information to assess air pollutant exposure during pregnancy is based on outdoor measurement of air pollutants (nitrogen dioxide [NO2], volatile organic compounds [VOC], ozone, particulate matter [PM10, PM2,5 ] and of their composition [polycyclic aromatic hydrocarbons]); measurement of indoor and personal exposure (VOC and NO2); urinary measurement of a biological marker of hydrocarbon exposure (1-hydroxypyrene); and data gathered by questionnaires and geographic information systems. These data allow individual air pollutant exposure indexes to be developed, which can then be used to analyze the possible effects of exposure on fetal development and child health. CONCLUSION: This protocol and the type of study allow an approximation to individual air pollutant exposure to be obtained. Finally, the large number of participants (N = 4,000), as well as their geographic and social diversity, increases the study's potential. PMID- 17419935 TI - [On the exercise of health authority: principles, regulations and uncertainty]. AB - Implementing health authority is a basic public health service. Part of the responsibility of public health managers is to ensure compliance with regulations. These are developed when certain risks are considered inadmissible. Mostly, the exercise of health authority deals with the routine application of detailed norms, although there is always some uncertainty, as shown by the frequent use of cautionary measures by health officers during inspections. However, epidemiologic surveillance periodically involves situations in which human health is damaged and there is no reference regulation; in these situations, health authorities must act according to their own criteria, weighing the risks of intervention against those of nonintervention. In this article, we present 3 such scenarios: using coercion in the treatment of patients with smear positive tuberculosis, regulation of activities with soy beans posing asthma risks, and setting limits to the professional activity of an HIV-positive physician. PMID- 17419936 TI - [Avoidable mortality and colorectal cancer]. PMID- 17419937 TI - [Criteria to define the list of causes of avoidable mortality: an unavoidable discussion]. PMID- 17419939 TI - Structural biology of the tumor suppressor p53 and cancer-associated mutants. AB - The tumor suppressor protein p53 is a transcription factor that plays a key role in the prevention of cancer development. In response to oncogenic or other stresses, the p53 protein is activated and regulates the expression of a variety of target genes, resulting in cell cycle arrest, senescence, or apoptosis. Mutation of the p53 gene is the most common genetic alteration in human cancer, affecting more than 50% of human tumors. Most of these mutations inactivate the DNA-binding domain of the protein. In this chapter, we describe the structure of the wild-type p53 protein and present structural and functional data that provide the molecular basis for understanding the effects of common cancer mutations. Further, we assess novel therapeutic strategies that aim to rescue the function of p53 cancer mutants. PMID- 17419940 TI - Immunotherapy by allogeneic stem cell transplantation. AB - During the past three decades, allogeneic stem cell transplantation (ASCT) has developed from being an experimental therapy in patients with endstage leukemia into a well-established therapy in patients with a range of disorders of the immunohematopoietic system. Graft-versus-host disease (GVHD), acute or chronic, attacking host tissue is a major threat. However, donor immunocompetent T cells have a potent graft-versus-leukemia effect. A combination of calcineurin inhibitors and methotrexate is the standard therapy to prevent GVHD. Modulation of the immunosuppressive regimen may induce mild acute and mild chronic GVHD, reduce the risk of relapse, and improve long-term survival. Natural killer cells also play a role in this context. Killer cell immunoglobulin-like receptor incompatibility between recipient and donor may reduce the risk of relapse in patients with myeloid leukemia. Relapse of leukemia is a major cause of death after ASCT. Minimal residual disease and recipient leukemia lineage-specific chimerism are sensitive techniques for early detection of leukemic relapse. Donor lymphocyte infusions can enhance the antitumor effect, especially for patients with molecular relapse. The allogeneic graft-versus-cancer effect has been demonstrated in patients with metastatic breast, renal, colorectal, ovarian, prostatic, and pancreatic carcinoma. Mesenchymal stem cells have immunomodulatory properties and may be used for immunomodulation of GVHD and tissue repair. All things considered, the future looks promising for ASCT. PMID- 17419941 TI - Mnt takes control as key regulator of the myc/max/mxd network. AB - Myc is the most frequently deregulated oncogene in human tumors. The protein belongs to the Myc/Max/Mxd network of transcriptional regulators important for cell growth, proliferation, differentiation, and apoptosis. The ratio between Mnt/Max and c-Myc/Max on the 5'-CACGTG-3' E-box sequence at shared target genes is of great importance for cell cycle progression and arrest. Serum stimulation of quiescent cells results in phosphorylation of Mnt and disruption of the critical Mnt-mSin3-HDAC1 interaction. This in turn leads to increased expression of the Myc/Mnt target gene cyclin D2. It is therefore possible that Myc function relies on its ability to overcome transcriptional repression by Mnt and that relief of Mnt-mediated transcriptional repression is of greater importance for regulation of target genes than the sole activation by Myc. In addition, Mnt has many features of a tumor suppressor and may thus be nonfunctional or inactivated in human tumors. In summary, accumulating evidence supports the model of Mnt as the key regulator of the network in vivo. PMID- 17419942 TI - Lytic cycle switches of oncogenic human gammaherpesviruses. AB - The seminal experiments of George and Eva Klein helped to define the two life cycles of Epstein-Barr Virus (EBV), namely latency and lytic or productive infection. Their laboratories described latent nuclear antigens expressed during latency and discovered several chemicals that activated the viral lytic cycle. The mechanism of the switch between latency and the lytic cycle of EBV and Kaposi's sarcoma-associated herpesvirus (KSHV) can be studied in cultured B cell lines. Lytic cycle activation of EBV is controlled by two viral transcription factors, ZEBRA and Rta. The homologue of Rta encoded in ORF50 is the lytic cycle activator of KSHV. Control of the lytic cycle can be divided into two distinct phases. Upstream events control expression of the virally encoded lytic cycle activator genes. Downstream events represent tasks carried out by the viral proteins in driving expression of lytic cycle genes and lytic viral DNA replication. In this chapter, we report three recent groups of experiments relating to upstream and downstream events. Azacytidine (AzaC) is a DNA methyltransferase inhibitor whose lytic cycle activation capacity was discovered by G. Klein and coworkers. We find that AzaC rapidly activates the EBV lytic cycle but does not detectably alter DNA methylation or histone acetylation on the promoters of the EBV lytic cycle activator genes. AzaC probably acts via a novel, yet to be elucidated, mechanism. The lytic cycle of both EBV and KSHV can be activated by sodium butyrate (NaB), a histone deacetylase inhibitor whose activity in disrupting latency was also discovered by G. Klein and coworkers. Activation of EBV by NaB requires protein synthesis; activation of KSHV is independent of protein synthesis. Thus, NaB works by a different pathway on the two closely related viruses. ZEBRA, the major downstream mediator of EBV lytic cycle activation is both a transcription activator and an essential replication protein. We show that phosphorylation of ZEBRA at its casein kinase 2 (CK2) site separates these two functions. Phosphorylation by CK2 is required for ZEBRA to activate lytic replication but not to induce expression of early lytic cycle genes. We discuss a number of unsolved mysteries about lytic cycle activation which should provide fertile territory for future research. PMID- 17419943 TI - No life without death. AB - Apoptosis-programed cell death-is the most common form of death in the body. Once apoptosis is induced, proper execution of the cell death program requires the coordinated activation and execution of multiple molecular processes. Here, we describe the pathways and the basic components of the death-inducing machinery. Since apoptosis is a key regulator of tissue homeostasis, an imbalance of apoptosis results in severe diseases like cancer, autoimmunity, and AIDS. PMID- 17419944 TI - Control of apoptosis in human multiple myeloma by insulin-like growth factor I (IGF-I). AB - Human multiple myeloma (MM) is characterized by the expansion of neoplastic plasmablasts/plasma cells with complex genetic aberrations and high dependence for survival and growth on cytokines produced in the bone marrow microenvironment. As tools in the study of MM about 80 authentic MM cell lines and a few relevant in vivo mouse models are available. The dependence on insulin like growth factor receptor (IGF-IR) signaling in the development and maintenance of the malignant phenotype in a variety of cancers is a rationale for attempts to improve tumor treatment by selectively inhibiting the IGF-IR in malignant cells by neutralizing antibodies, dominant negative IGF-IR, and IGF-IR siRNA. Testing the hypothesis that abrogating IGF-IR-mediated signaling of survival should make MM cells more susceptible to apoptosis, our studies have so far provided proof-of principle by the demonstration that inhibition of a signaling pathway stimulating survival renders cells susceptible to drug-induced apoptosis when the drug (dexamethasone) and inhibitor (rapamycin) converge on the same target, that is p70(S6K). The recent publication of the three-dimensional structure of the IGF-IR kinase domain has facilitated the development of IGF-IR inhibitors of the cyclolignan family, that is picropodophyllin, with capacity to distinguish also in vivo between the IGF-IR and the insulin receptor. Studies in vitro and in vivo with picropodophyllin show promising effects, that is apoptosis induction and growth arrest, and have made it possible to evaluate the biological and therapeutic effects of inhibition of the IGF-IR signaling in MM. PMID- 17419945 TI - c-MYC impairs immunogenicity of human B cells. AB - Deregulation of c-myc expression through chromosomal translocation is essential in the pathogenesis of Burkitt's lymphoma (BL). A characteristic feature of BL cells, compared to Epstein-Barr Virus (EBV)-immortalized B cells, is their lack of immunogenicity. To study the contribution of EBV genes and of the c-MYC protein to this phenotype, we have generated a conditional B cell system in which the viral proliferation program and expression of c-myc can be regulated independently of each other. In cells proliferating due to exogenous c-myc overexpression, the cell surface phenotype, the pattern of proliferation in single cell suspension, and the immunological characteristics of BL cells could be completely recapitulated. Yet, it had remained open whether nonimmunogenicity is the default phenotype when EBNA2 and LMP1 are switched off, or whether c-MYC actively contributes to immunosuppression. We provide evidence also for the latter by showing that c-MYC down-regulates genes of the NF-kappaB and interferon pathway in a dose-dependent fashion. c-MYC acts at at least two different levels, the level of interferon induction as well as at the level of action of type I and type II interferons on their respective target promoters. c-MYC does not block the interferon pathway completely, it shifts the balance and increases the threshold of interferon induction and action. PMID- 17419946 TI - Cancer dormancy: lessons from a B cell lymphoma and adenocarcinoma of the prostate. AB - Cancer dormancy delineates a situation in which residual tumor cells persist in a patient with no apparent clinical symptoms. Although the precise mechanisms underlying cancer dormancy have not been explained, experimental models have provided some insights into the factors that might be involved in the induction and maintenance of a tumor dormant state. The authors of the present chapter studied a murine B cell lymphoma that can be made dormant when interacting with antibodies directed against the idiotype on its immunoglobulin Ig receptor. This experimental model of antibody-induced dormancy enabled the isolation and characterization of dormant lymphoma cells. The results indicated that anti-Ig antibodies activate growth-inhibiting signals that induced cycle arrest and apoptosis. This process appeared to be balanced by the growth of the tumor cells such that the tumor did not expand. In contrast, antibodies against HER 2expressed on prostate adenocarcinoma (PAC) cells were not growth inhibitory. However, an immunotoxin (IT) prepared by conjugating HER-2 to the A-chain of ricin (RTA) was internalized by PAC cells, followed by induction of cycle arrest and apoptotic death. Infusion of HER-2-specific IT into PAC-bearing immunodeficient mice did not eradicate the tumor but retained it dormant over an extended period of time. Hence, certain aspects of signaling receptors expressed on cancer can be manipulated by antibodies to induce and maintain a tumor dormant state. PMID- 17419947 TI - Therapeutic targets of multiple angiogenic factors for the treatment of cancer and metastasis. AB - Like any growing healthy tissues, tumors build up their blood vessels by three mechanisms: angiogenesis, vasculogenesis, and intersucception. Vascular endothelial growth factor-A (VEGF-A) is one of the key factors responsible for stimulation and maintenance of the disorganized, leaky, and torturous tumor vasculature. In addition to VEGF-A, tumors produce multiple other factors to stimulate blood vessel growth. These include members in the platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), VEGF-C, insulin-like growth factor (IGF), angiopoietin (Ang), and hepatocyte growth factor (HGF) families. Recent studies show that these angiogenic factors can also promote lymphangiogenesis and potentially lymphatic metastasis. Understanding the roles of individual and combined angiogenic factors in promoting tumor angiogenesis is crucial for defining therapeutic targets and antiangiogenic drug development for the treatment of cancer. PMID- 17419948 TI - Novel three-dimensional organotypic liver bioreactor to directly visualize early events in metastatic progression. AB - Metastatic seeding leads to most of the morbidity from carcinomas. However, little is known of this key event as current methods to study the cellular behaviors utilize nonrepresentative in vitro models or follow indirect subsequent developments in vivo. Therefore, we developed a system to visualize over a multiday to multiweek period the interactions between tumor cells and target organ parenchyma. We employ an ex vivo microscale perfusion culture system that provides a tissue-relevant environment to assess metastatic seeding behavior. The bioreactor recreates many features of the fluid flow, scale, and biological functionality of a hepatic parenchyma, a common site of metastatic spread for a wide range of carcinomas. As a test of this model, prostate and breast carcinoma cells were introduced. Tumor cell invasion and expansion could be observed by two photon microscopy of red fluorescent protein (RFP)- and CellTracker-labeled carcinoma cells against a green fluorescent protein (GFP)-labeled hepatic tissue bed over a 14-day period. Tumors visible to the naked eye could be formed by day 25, without evident necrosis in the >0.3-mm tumor mass. These tumor cells failed to grow in the absence of the supporting three-dimensional (3D) hepatic microtissue, suggesting paracrine or stromal support function for the liver structure in tumor progression. Initial ultrastructural studies suggest that early during the tumor-parenchyma interactions, there are extensive interactions between and accommodations of the cancer and host cells, suggesting that the tumor-related epithelial-mesenchymal transition (EMT) reverts, at least transiently, to promote metastatic seeding. In sum, our 3D ex vivo organotypic liver tissue system presents a critical vehicle to examine tumor-host interactions during cancer metastasis and/or invasion. It also circumvents current limitations in assays to assess early events in metastasis, and provides new approaches to study molecular events during tumor progression. PMID- 17419949 TI - PDGF receptors as targets in tumor treatment. AB - Signaling through platelet-derived growth factor (PDGF) receptors contributes to multiple tumor-associated processes. The recent introduction of clinically useful PDGF inhibitors have the last years validated PDGF receptors in malignant and stromal cells as relevant cancer drug targets. Mutational activation of PDGF receptor signaling in malignant cells has been described in some rare tumor types such as dermatofibrosarcoma protuberans, a subset of GISTs, and some hematologic malignancies. Furthermore, expression of PDGF receptors on pericytes is a common characteristic of solid tumors. The clinical efficacy of novel multikinase inhibitors, such as sunitinib and sorafenib, most likely involves targeting of PDGF receptor-dependent pericytes. Preclinical studies suggest that targeting of stromal PDGF receptors might also constitute a novel strategy to enhance tumor drug uptake. Finally, recent studies have implied both pro- and antimetastatic effects of PDGF receptors on malignant and stromal cells. The studies on the roles of PDGF receptors in cancer signaling are thus presently in a dynamic phase where collaborations between oncologists, pathologists, and tumor biologists are predicted to be highly productive. PMID- 17419951 TI - Targeted cancer therapy: promise and reality. AB - Signal transduction therapy for cancer targets specific molecular elements that are essential for survival of the tumor. Gleevec has a profound effect on early phase chronic myeloid leukemia because it inhibits the major driving factor of the tumor, BCR-ABL. Almost all other cancers depend on several factors, and blocking a single signal transduction factor is largely ineffective. Effective signal transduction therapy will entail finding the appropriate combination of signal transduction inhibitors for each cancer. We discuss the use of preclinical animal models to predict successful signal transduction therapy in the clinic, and conclude that their utility is limited. PMID- 17419950 TI - Extracellular matrix, nuclear and chromatin structure, and gene expression in normal tissues and malignant tumors: a work in progress. AB - Almost three decades ago, we presented a model where the extracellular matrix (ECM) was postulated to influence gene expression and tissue-specificity through the action of ECM receptors and the cytoskeleton. This hypothesis implied that ECM molecules could signal to the nucleus and that the unit of function in higher organisms was not the cell alone, but the cell plus its microenvironment. We now know that ECM invokes changes in tissue and organ architecture and that tissue, cell, nuclear, and chromatin structure are changed profoundly as a result of and during malignant progression. Whereas some evidence has been generated for a link between ECM-induced alterations in tissue architecture and changes in both nuclear and chromatin organization, the manner by which these changes actively induce or repress gene expression in normal and malignant cells is a topic in need of further attention. Here, we will discuss some key findings that may provide insights into mechanisms through which ECM could influence gene transcription and how tumor cells acquire the ability to overcome these levels of control. PMID- 17419952 TI - Restoration of wild-type p53 function in human tumors: strategies for efficient cancer therapy. AB - The p53 tumor suppressor gene is mutated in around 50% of all human tumors. Most mutations inactivate p53's specific DNA binding, resulting in failure to activate transcription of p53 target genes. As a consequence, mutant p53 is unable to trigger a p53-dependent biological response, that is cell cycle arrest and apoptosis. Many tumors express high levels of nonfunctional mutant p53. Several strategies for restoration of wild-type p53 function in tumors have been designed. Wild-type p53 reconstitution by adenovirus-mediated gene transfer has shown antitumor efficacy in clinical trials. Screening of chemical libraries has allowed identification of small molecules that reactivate mutant p53 and trigger mutant p53-dependent apoptosis. These novel strategies raise hopes for more efficient cancer therapy. PMID- 17419953 TI - Evidence synthesis combining individual patient data and aggregate data: a systematic review identified current practice and possible methods. AB - OBJECTIVE: Meta-analysis of individual patient data (IPD) is the "gold-standard" for synthesizing evidence across several studies. Some studies, however, may only provide aggregate data (AD). In this situation researchers might need to combine IPD with AD to utilize all the evidence available. Here, we review applied IPD meta-analysis articles to assess if and how AD is combined with IPD in practice. STUDY DESIGN AND SETTING: A systematic review of articles identified from bibliographic databases and searches. RESULTS: We identified 33 applied IPD articles that combined IPD and AD and 166 that did not. For each article, we recorded the proportion of total studies providing IPD, and found that articles combining IPD and AD had, on average, IPD available in only 64% of studies (compared to 90% in articles not combining IPD and AD). Two different methods were used to combine IPD and AD, the two-stage method and analysis of partially reconstructed IPD, but a review of methodological articles identified two further methods, multilevel modeling and Bayesian hierarchical related regression. We summarize each method to aid practitioners. CONCLUSION: Combining IPD and AD is a relevant issue for evidence synthesis, and the further development and validation of suitable meta-analysis methods is needed. PMID- 17419954 TI - A combined-method psychometric analysis recommended modification of the multidimensional health locus of control scales. AB - OBJECTIVE: The Multidimensional Health Locus of Control (MHLC) scales are widely used to measure beliefs about determinants of persons' health. We evaluated the scales over the largest-ever disease-specific sample of subjects using a combined method psychometric approach. STUDY DESIGN AND SETTING: We performed a secondary analysis of data from 1,206 subjects from three osteoarthritis studies, using Rasch analysis and confirmatory factor analysis simultaneously. Differential item functioning (DIF) by gender and data source, scale dimensionality, and item fit were examined. The Rasch model fit the data if Rasch residual principal components analysis (PCA) corroborated three distinct dimensions and item fit statistics fell between 0.80 and 1.20. The confirmatory factor (CFA) model fit the data if factor loadings exceeded 0.50 for all items. RESULTS: DIF by gender or data source was not materially evident for any items. PCA supported existence of three dimensions in the data. Both Rasch and CFA models fit the data for 16 items; two items were detected as misperforming. When these items were removed, fit of both models improved. CONCLUSION: Results of this large-sample evaluation of the MHLC scales corroborated earlier findings that removal of certain items improves the scales. The combined Rasch-CFA approach provided better insight to scale performance problems than either method alone provided. PMID- 17419955 TI - Three methods for minimally important difference: no relationship was found with the net proportion of patients improving. AB - OBJECTIVE: To determine the impact on a responder type analysis of using three published methods to obtain the minimally important difference (MID) on the conclusion of a randomized controlled trial (RCT). STUDY DESIGN AND SETTING: Using data from an RCT of supportive-expressive group therapy (SEGT-intervention) vs. standard care (control) in women with metastatic breast cancer, we measured individual responsiveness to change according to three levels of predefined MID (0.2 SD, 0.5 SD, and 1 standard error of measurement) of the following six validated questionnaires: Profile of Mood States, Impact of Event Scale, Psychosocial Adjustment to Illness Scale, EORTC Quality-of-Life Questionnaire Core-30, Mental Adjustment to Cancer, and a pain visual analog scale. The proportion of women improved by SEGT and the number needed to treat according to three levels of MID were calculated. RESULTS: There was no consistent difference in the net proportion of women improving with the SEGT vs. control arm according to the three different levels of MID. CONCLUSION: The choice between different levels of distribution-based MID did not make an important difference in the net proportion of women improving with the SEGT. Future research should compare MID derived from clinical anchors, in particular patient opinions. PMID- 17419956 TI - A randomized trial of Pegaptanib sodium for age-related macular degeneration used an innovative design to explore disease-modifying effects. AB - OBJECTIVE: Effectively evaluating disease-modifying effects in clinical trials has posed a problem for clinical trialists and drug development. One method that has been proposed to evaluate disease-modifying effects has been the re randomization of active group participants to discontinue the intervention after a period sufficient to produce therapeutic effects. We aimed to determine if this design would permit inferences regarding disease modification in a trial evaluating Pegaptanib sodium, an intra-ocular injection, for the treatment of age related macular degeneration. STUDY DESIGN AND SETTING: In two identically designed trials, 1,186 patients were randomized to receive 54 weeks of treatment. After 54 weeks, 1,053 were re-randomized to either stay on treatment or discontinue treatment. Patients were seen at multicenter outpatient clinics. RESULTS: We found that patients randomized to discontinue treatment after 54 weeks of treatment and followed for a further 48 weeks, were significantly different than the control group (sham) in loss of 15 letters of vision (Relative Risk 0.70, 95% Confidence Interval 0.57-0.88, P=0.002), indicating that treatment is disease modifying. This effect was consistent throughout our sensitivity analyses. CONCLUSION: This trial is the first example of a clinical trial evaluating disease-modifying effects and this design should influence drug discovery to determine further therapeutic potential of pharmacologic interventions. PMID- 17419957 TI - Generalized approaches to partitioning the attributable risk of interacting risk factors can remedy existing pitfalls. AB - OBJECTIVE: To clarify the properties of different approaches to estimate the contribution of single-risk factors to the disease load in a population. STUDY DESIGN AND SETTING: Three methods of partitioning attributable risks are reviewed and two additional procedures as modifications of the existing algorithms are introduced. Basis properties of the approaches are outlined in the simplest setting with two exposure variables. The extension to more complex settings is illustrated by an example involving three risk factors. RESULTS: The quantification of the impact of single-risk factors can vary considerably according to the method used. Different orderings of the risk factors with respect to their impact can occur. Approaches can be classified according to two features: (i) inclusion or exclusion of partial interactions between risk factors, (ii) equal or proportional distribution of the surplus resulting from the combined action of risk factors. Practical applications have to carefully consider intrinsic limitations of all partitioning approaches. CONCLUSION: The decision on which concept to use when partitioning attributable risks on the population level should be based on the desired properties the solution ought to have. Arguments from game-theoretical reasoning can help to guide further research in this area, especially in exploring the methods using proportional division rules that are not yet fully understood. PMID- 17419958 TI - Children proxies' quality-of-life agreement depended on the country using the European KIDSCREEN-52 questionnaire. AB - OBJECTIVE: The aims of this study were to assess the level of agreement and the magnitude of discrepancies between children and their parents, and whether these levels of agreement/discrepancy depend on the country of living, the domains of quality of life assessed, the age and gender of the children, and other background variables. STUDY DESIGN AND SETTING: The KIDSCREEN pilot study involved 2,526 youth-proxy pairs in seven European countries. The health-related quality of life (HRQoL) of children and their parents as proxy has been assessed in parallel using a 10-dimension KIDSCREEN-52 Pilot test questionnaire. RESULTS: For the 10 dimensions, the mean difference between proxy and youth score decreases as the HRQoL level increases. Physical and cognitive domains showed the major agreement between youth and proxy measure. Social and psychological domains presented the main discrepancies. Linear regression models highlighted that the agreement was depending on the country for the 10 dimensions. Impact of age and gender on agreement were not consistent across the 10 dimensions. CONCLUSION: Agreement is higher for the girls than for the boys and for the adolescents than for children. To further explore the country effect on agreement, results need replication in representative studies. PMID- 17419959 TI - Racial/ethnic differences in preferences for the EQ-5D health states: results from the U.S. valuation study. AB - OBJECTIVE: We examined differences in preferences for the EQ-5D health states among blacks, Hispanics, and others living in the United States. STUDY DESIGN AND SETTING: A multi-stage probability sample was selected from the adult U.S. population. Each respondent valued a subset of the 243 EQ-5D health states. Regression analysis was used to analyze differences in mean valuations for 13 health states among the racial/ethnic groups. First, we compared unadjusted mean valuations among the three groups. Second, we evaluated differences in mean valuations among the groups after adjusting for other sociodemographic characteristics. Third, we evaluated the impact of race/ethnicity on estimates derived from the U.S. health state valuation (i.e., D1) model. RESULTS: Valuations differed among the groups for seven of the 13 health states, and these differences persisted after adjusting for other sociodemographic factors. Blacks appeared to perceive extreme health problems to be associated with less disutility than did members of the other two groups. CONCLUSION: Within the United States, there exist racial/ethnic differences in the perceived desirability of the EQ-5D health states that cannot be readily explained by socioeconomic disparities. PMID- 17419960 TI - External validation of prognostic models for critically ill patients required substantial sample sizes. AB - OBJECTIVE: To investigate the behavior of predictive performance measures that are commonly used in external validation of prognostic models for outcome at intensive care units (ICUs). STUDY DESIGN AND SETTING: Four prognostic models (Simplified Acute Physiology Score II, the Acute Physiology and Chronic Health Evaluation II, and the Mortality Probability Models II) were evaluated in the Dutch National Intensive Care Evaluation registry database. For each model discrimination (AUC), accuracy (Brier score), and two calibration measures were assessed on data from 41,239 ICU admissions. This validation procedure was repeated with smaller subsamples randomly drawn from the database, and the results were compared with those obtained on the entire data set. RESULTS: Differences in performance between the models were small. The AUC and Brier score showed large variation with small samples. Standard errors of AUC values were accurate but the power to detect differences in performance was low. Calibration tests were extremely sensitive to sample size. Direct comparison of performance, without statistical analysis, was unreliable with either measure. CONCLUSION: Substantial sample sizes are required for performance assessment and model comparison in external validation. Calibration statistics and significance tests should not be used in these settings. Instead, a simple customization method to repair lack-of-fit problems is recommended. PMID- 17419961 TI - Study of observed and self-reported HRQL in older frail adults found group-level congruence and individual-level differences. AB - OBJECTIVE: To evaluate the consistency of self-reported health-related quality of life (HRQL) using the Health Utilities Index Mark 2 (HUI2) with observer rated HRQL using the Minimum Data Set Health-Status Index (MDS-HSI). STUDY DESIGN AND SETTING: Frail older home care clients in Calgary Alberta and Wayne County, Michigan responded to HUI2 questionnaires and were assessed using the Minimum Data Set Home Care tool (n=514). HRQL scores were calculated and compared for the HUI2 and the MDS-HSI. The intraclass correlation coefficient (ICC) was used to assess individual level agreement. RESULTS: The MDS-HSI provided HRQL scores that consistently averaged 0.10 points higher than HUI2 self-reported HRQL scores overall and within client characteristics. The ICC was 0.46 in the full population but increased to 0.63 when 10% of the sample with the largest discrepant scores was removed. Pain and emotion health attributes showed the lowest level of agreement. CONCLUSION: The MDS-HSI and HUI2 provide analogous group-level results but only moderate individual-level agreement. When HUI2 survey data are not available, the MDS-HSI can be used to substitute for the HUI2 in group-level comparisons but not for individual clinical evaluation comparisons. PMID- 17419962 TI - Low agreement was found between pharmacy data and physician reported use of hospital-administered antenatal corticosteroids. AB - OBJECTIVE: To evaluate physicians' recall accuracy on starting year of routine use of antenatal corticosteroids (ACS) to women facing imminent preterm delivery. STUDY DESIGN AND SETTING: Starting year of routine treatment with ACS was evaluated by a questionnaire mailed to all 52 Swedish maternity wards. The information was compared with that obtained from a telephone interview with physicians involved in the introduction of routine ACS and with pharmacy data. From pharmacy data, routine use of ACS was defined as >or=20 mg betamethasone or dexamethasone purchased per maternity ward, year, and preterm delivery. RESULTS: 24 hospitals with >or=10 preterm deliveries/year, having started ACS 1976-1997 and with information from questionnaire, interview, and pharmacy data were included in the analyses. There was fair agreement (kappa=0.38, P<0.001) on starting year between questionnaires and telephone interviews with physicians and slight agreement (kappa=0.06, P=0.098) between questionnaires and pharmacy data. Three hospitals had complete agreement on starting year according to questionnaire, telephone interview, and pharmacy data. CONCLUSION: Agreement between information obtained from questionnaires, telephone interviews with physicians, and pharmacy data on use of ACS was low. Physician reported past drug use needs to be validated and pharmacy data can be useful for such purpose. PMID- 17419963 TI - Physical activity prevented functional decline among frail community-living elderly subjects in an international observational study. AB - OBJECTIVES: The aims of the study were to describe the prevalence of physical activity in a sample of older adults in home care in Europe and to examine the relationship between physical activity and incident disability. STUDY DESIGN AND SETTING: Study population consisted of a random sample of 2,005 subjects aged 65 or older admitted to home care programs in 11 European Home Health Agencies who participated in AgeD in HOme Care project. Participants who reported spending 2 or more hours of physical activities in last 3 days were defined physically active. Disability performing activities of daily living was defined as the need of assistance in one or more of the following ADL: eating, dressing, transferring, mobility in bed, personal hygiene, and toileting. RESULTS: More than 50% of participants were physically active. During a median follow-up of 12 months, 370 subjects (15%) became disabled. After adjusting for age, gender, and other possible confounding variables, active subjects were significantly less likely to become disabled compared to those reporting no or very low-intensity physical activity (OR, 0.67; 95% CI 0.53-0.84). CONCLUSIONS: These findings support the possibility that physical activity has an independent effect on functional autonomy among frail and old people. PMID- 17419964 TI - Self-reported information and pharmacy claims were comparable for lipid-lowering medication exposure. AB - OBJECTIVE: To examine agreement between self-reported exposure to lipid-lowering medications and objective evidence of filling prescribed lipid-lowering medications. STUDY DESIGN AND SETTING: Using data from 7,918 adults from the Adverse Childhood Experiences (ACE) Study, we calculated the sensitivity, specificity, and positive (PV+) and negative (PV-) predictive values, and likelihood ratios for self-reported exposure to lipid-lowering medications compared to exposure obtained from pharmacy claims (gold standard) both overall and by age, sex, race/ethnicity, education, and ACE Score. RESULTS: Eight percent (n=655) of adults self-reported lipid-lowering medication exposure, and 379 adults filled at least one lipid-lowering prescription within 60 days of the baseline exam during 1997. The sensitivity of self-reported exposure was nearly 94%; the specificity was 96%; the PV+ was 54%; and the PV- was nearly 100%. Values for sensitivity, specificity, PV+, and PV- were similar across participant characteristics. CONCLUSION: A self-reported measure of lipid-lowering medication exposure was accurate with high sensitivity and specificity while the PV+ of self reported lipid-lowering medication exposure was relatively low. These findings suggest that self-reported exposure to lipid-lowering medications may be useful in surveys that examine the prevalence of hyperlipidemia, but may overestimate actual exposure in studies monitoring trends in use of lipid-lowering medications. PMID- 17419965 TI - Studies reporting ROC curves of diagnostic and prediction data can be incorporated into meta-analyses using corresponding odds ratios. AB - OBJECTIVE: To develop an approach by which studies describing the accuracy of diagnostic tests or clinical predictions can be combined in a meta-analysis, even though studies may report their results using different summary measures. STUDY DESIGN: A method is proposed to allow algebraic and numerical conversion of values of the Receiver Operating Characteristic Area Under the Curve (AUC) summary statistic into corresponding odds ratios (OR). A similar conversion is demonstrated for the standard errors (SEs) of these summary statistics. RESULTS: The conversion of the AUC values into OR values was achieved using a logit threshold model. The delta method was used to convert the associated SEs. An example concerning predictions of mortality in the intensive care unit illustrates the calculations. CONCLUSION: This paper provides an accessible method that permits the meta-analyst to overcome some of the difficulties implied by incomplete and inconsistent reporting of research studies in this area. It allows all studies to be included on the same metric, which in turn more easily permits exploration of issues such as heterogeneity. The method can readily be used for meta-analyses of diagnostic or screening tests, or for prediction data. PMID- 17419966 TI - Submissions of scientific papers should not become a sophistication. PMID- 17419967 TI - Placebo surgery research: a blinding imperative. PMID- 17419972 TI - Mucus structure and properties in cystic fibrosis. AB - The biophysical properties of airway secretions are largely determined by the polymeric components. In normal airway mucus, the gel-forming mucins, MUC5AC and MUC5B, are responsible not only for the viscoelastic properties essential for clearance and protecting the airway epithelium from invaders and water evaporation. With chronic airway infection, inflammatory cell necrosis leads to a predominance of polymeric DNA and F-actin. There is almost no mucin in the sputum of patients with established cystic fibrosis lung disease. Sputum viscoelastic and surface properties determine how well secretions can be cleared by cilia or cough. In this mini-Symposium on Airway Clearance in Cystic Fibrosis, the physiology of CF secretion structure and rheology is discussed in the context of medications and physical maneuvers for enhancing sputum clearance. PMID- 17419973 TI - Chest physical therapy, breathing techniques and exercise in children with CF. AB - Chest physiotherapy in the form of airway clearance techniques and exercise has played an important role in the treatment of cystic fibrosis. Until the 1990s the primary airway clearance technique used was postural drainage combined with percussion and vibration (PD&P). It was introduced into the treatment of CF with little evidence to support its efficacy and once established, it has been difficult ethically to perform a study comparing PD&P to no treatment. A common question, yet unanswered is when should it be commenced, especially for the newly diagnosed asymptomatic CF patient? Recently, the technique of PD&P has been modified to include only non-dependant head-down positioning due to the detrimental effects of placing a person in a Trendelenburg position. In the 1990s other airway clearance techniques gained popularity, in that they could be performed independently, in a sitting position and avoided many of the detrimental effects of PD&P. These techniques include the Active cycle of breathing technique, formally called the Forced expiration technique and Autogenic drainage. Both these breathing techniques aim at using expiratory airflow to mobilize secretions up the airways and incorporate breathing strategies to assist in the homogeneity of ventilation. Studies suggest that both these techniques are as effective if not more effective than as PD&P and offer many advantages over PD&P. It has been suggested that exercise can be used as an airway clearance technique; however the literature does not support this. Rather, when exercise is used in addition to an airway clearance technique there is enhanced secretion removal and an overall benefit to the patient. Further research needs to be directed at assessing the effects of an airway clearance technique on the individual patient using appropriate outcome measures. PMID- 17419974 TI - Airway clearance devices in cystic fibrosis. AB - Clearance of infected airway secretions is essential to preserve lung function in patients with cystic fibrosis (CF). Although the value of regular airway clearance treatments has been shown in many studies, adherence to the prescribed treatments is not very good (see Making airway clearance successful, pp. 000 000). In the past the only method available was conventional chest physiotherapy (CCPT; also known as manual percussion and postural drainage). CCPT remains the 'gold standard' of airway clearance methods and may be the best choice for some patients, such as infants and young children. However, the many newer methods of airway clearance available now allow CF patients and their families to choose the techniques and devices that best suits them. Most of the newer airway clearance devices have been studied in comparison to standard chest physiotherapy and most studies show no advantage of one method over another. This review will describe newer airway clearance devices available for CF patients and discuss evidence for the effectiveness of these devices compared to standard chest physiotherapy. PMID- 17419975 TI - Mucolytics in cystic fibrosis. AB - Mucus accumulation in the lower airways is a key feature of cystic fibrosis (CF) lung disease. The major component of mucus in CF is not mucin derived from mucus producing cells but rather pus that includes viscous material such as polymerized DNA derived from degraded neutrophils. This has important implications for mucolytic therapy aiming to improve mucus clearance from the airways, since degradation of mucin may not be a suitable treatment strategy. In addition, thinning of secretions may not always be beneficial, since it may negatively affect certain aspects of mucus transport such as cough clearance. While inhaled N-acetylcysteine has been used as a mucolytic drug in CF for decades, there is little evidence that it has any beneficial effect. Dornase alfa has been shown to reduce pulmonary exacerbations and improve lung function and is currently the only mucolytic agent with proven efficacy in CF. Newer agents targeting other components of CF mucus, such as filamentous actin, are currently in development. Ultimately, drugs that are mucokinetic, which preserve viscoelasticity, rather than mucolytic may prove to be beneficial for CF lung disease in the future. PMID- 17419976 TI - Other mucoactive agents for cystic fibrosis. AB - This review examines specific mucoactive agents from three classes: expectorants, which add water to the airway; ion-transport modifiers, which promote ion and water transport across the epithelium of the airway; and mucokinetics, which improve cough-mediated clearance by increasing airflow or reducing sputum adhesivity. The agents are isotonic and hypertonic saline, mannitol, denufosol and beta-agonists. Our understanding of these agents has recently improved through pre-clinical research, clinical trials and, in particular, extensive research into the nature of the liquid lining the surface of the airway, both in health and in cystic fibrosis (CF). For each agent, recent research is reviewed, highlighting the evidence for possible mechanisms of action and for clinical efficacy in CF, as well as the implications for the optimal clinical application of the agent. PMID- 17419977 TI - Making airway clearance successful. AB - Airway clearance therapy (ACT) requires a substantial time, effort and resource commitment on the part of families and patients with cystic fibrosis (CF). Lack of compliance with caregiver recommendations and adherence to a proposed therapeutic plan occurs more than 50% of the time with ACT and is particularly common in adolescents. There is a large menu of ACT devices and techniques from which to choose. Increasing adherence and compliance requires an effort on the part of the CF team, including regular visits to the CF center, matching the patient to the device or technique based on lifestyle and disease severity, selecting appropriate outcome measures and monitoring adherence and compliance. Successful ACT also requires the CF team to understand individual patient situations, individualize ACT programs and accept a realistic level of patient participation. PMID- 17419978 TI - Pulmonary complications of haematopoietic cell transplantation in children. AB - Paediatric haematopoietic cell transplantation has experienced significant advances in the last few decades. However, pulmonary complications are an important limitation to the efficacy of this intervention, contributing to post transplantation morbidity and mortality. Such complications persist even in experienced centres and occur in adult and paediatric recipients. This review identifies the paediatric pulmonary complications that are commonly seen following haematopoietic cell transplantation and addresses both infectious and non-infectious aetiologies and their clinical manifestations, evaluation, and potential therapy. Ultimately, improvement in outcomes will require attention to immunosuppression as well as traditional diagnostic procedures and treatment. This article aims to review the current state of pulmonary complications post transplantation, to examine the impact of our recent advances and changes in treatment, and to identify potential future therapies and hypothesise what role these might have on long-term survival. PMID- 17419979 TI - Toxicological aspects and applications of nanoparticles in paediatric respiratory disease. AB - Most research in the area of micro- and nano-particles as applied to respiratory disease has been on potential toxic effects. Particulate emissions from industrial processes, coal burning and diesel exhaust have been shown to cause a variety of adverse effects both in vitro and in vivo. However, the vast majority of these studies has focused on larger, micron-sized particles. It is only within the last few years that the emphasis has shifted to nanoparticles as nanotechnology research and its applications have increased. Investigations have also begun into how nanoparticles may be used for therapeutic and imaging purposes in pulmonary diseases such as tuberculosis and cystic fibrosis. Some of these applications, along with recent studies on the toxic effects of nanoparticulate emissions will be reviewed in this article. PMID- 17419980 TI - Congenital cyst adenomatoid malformations: resect some and observe all? AB - With the advent of improved antenatal imaging over the past 10 years, the diagnosis, assessment and management of congenital cystic lung abnormalities have changed. These were once considered the exclusive domain of the surgeon, who had the authority to operate on all congenital cystic lung abnormalities regardless of size or clinical signs in order to avoid the risk of cancer and improve lung growth in even asymptomatic infants. Clinicians are reconsidering this approach in the light of an appreciation of the spontaneous improvement and possible resolution that occurs over months to years with many of these lesions. The risks of subsequent cancer are poorly understood and probably overstated, whereas the magnitude of compensatory lung growth is poorly defined in the majority of children with small unilateral congenital cystic abnormalities. The evolution of regional, national and ideally international databases will provide much-needed longitudinal data to better inform clinicians of the optimal way in which to manage these children. PMID- 17419981 TI - Lung abscess in children. AB - Lung abscess is an uncommon paediatric problem, with a paucity of quality data on the subject in the medical literature. Although the condition has for many years been managed successfully with prolonged courses of intravenous antibiotics, the evolution of interventional radiology has seen the use of percutaneously placed 'pigtail catheters' inserted under ultrasound and computed tomography guidance become the mainstay of therapy where such resources are available. This has been suggested to result in a more rapid defervescence of fever and symptoms, shorter periods of intravenous antibiotics and a decreased length of inpatient care. More invasive procedures, aspiration and drainage, together with improved microbiological diagnostic techniques, including polymerase chain reaction testing, has increased the yield of pathogens identified from abscess fluid samples. Culture results will guide treatment, especially for immunocompromised subjects who may develop a lung abscess as a complication of their underlying condition. The predominant pathogens isolated from primary lung abscesses in children include streptococcal species, Staphylococcus aureus and Klebsiella pneumoniae. Children with a lung abscess, both primary and secondary, have a significantly better prognosis than adults with the same condition. PMID- 17419982 TI - What's new in paediatric sleep? AB - The major recent advance in our understanding of paediatric sleep is the publication of reference values for sleep in children aged 3.2-8.6 years. These data show developmental changes reflecting a subtle process of the maturation of the central nervous system with regard to sleep in childhood. In infants, a significant negative correlation has been observed between a snore-associated arousal index and an infant development scale, underlining that snoring is less innocent than has been suggested. A link between obstructive sleep apnoea (OSA) and airway inflammation has been demonstrated, with children with OSA having significantly higher expression of the leukotriene (LT) 1 and 2 receptors and higher concentrations of LT C4/D4/E4 and LT B4 in adenotonsillar tissues than children with recurrent rhinitis who have no OSA. This explains the efficacy of treatment for OSA with montelukast, a LT receptor antagonist, alone or in combination with corticosteroids. By using peripheral arterial tonometry, a noninvasive technique that allows the moment-to-moment measurement of sympathetic tone, persistent waking-associated autonomic nervous system dysfunction has been demonstrated in young children with sleep-disordered breathing (SDB). As such, SDB in childhood may represent a cardiovascular risk factor in adulthood. PMID- 17419983 TI - Measurement of maximal pressures and the sniff manoeuvre in children. AB - Maximal static inspiratory and expiratory pressures are simple, noninvasive tests that evaluate global inspiratory and expiratory muscle strength. But these tests may be difficult or impossible to perform in young children. The sniff is a natural maneuver which many children find easier to perform than maximal pressures. The measurement of the nasal inspiratory pressure represents a valuable inspiratory muscle test which allows the extension of inspiratory muscle testing to a younger and larger paediatric population. PMID- 17419984 TI - Fractional exhaled nitric oxide and induced sputum. AB - Conventional asthma management aimed at controlling the underlying airway inflammation is classically based on symptoms and lung function. More recently, various non invasive markers of airway inflammation have become available. The ideal measurement method should be safe, non-invasive, easy to perform, reproducible and accurate. Fractional exhaled nitric oxide (FeNO) measurements fulfil these criteria, however some issues concerning cut off values and clinical variability as well as the interpretation of high values in the absence of symptoms still need to be solved. Induced sputum measurements are more labour intensive, however have the advantage of providing direct, additional information on the current inflammatory status of the airways. The most frequently analysed marker is sputum eosinophil percentage, although other markers of inflammation have also been under investigation. Both methods, FeNO and induced sputum should be seen as complementary to the conventional tools such as spirometry and bronchial hyperresponsiveness (BHR) testing. PMID- 17419985 TI - The centrosome opens the way to mitosis. AB - During mitosis, the interaction between chromosomes and microtubules requires nuclear envelope disassembly in prophase. Two articles in this issue of Developmental Cell show that centrosomes have a role in promoting nuclear envelope breakdown (Hachet et al., 2007; Portier et al., 2007). Surprisingly, the role of the centrosome in this process is independent of its role as a microtubule nucleation organelle. Instead, the centrosome seems to act as a spatial regulator for the activation of the Aurora A kinase. PMID- 17419986 TI - Timing is everything: regulation of Cdk1 and aneuploidy. AB - The spindle checkpoint ensures the proper partition of the chromosomal content of dividing cells, by controlling the transition from metaphase to anaphase. In a recent issue of Cancer Cell, Vecchione and coworkers report that the protein product of the tumor suppressor gene Lzts1 (Leucine zipper tumor suppressor-1) binds the Cdk1 phosphatase Cdc25C and stabilizes it by protecting it from proteasomal degradation (Vecchione et al., 2007). Partial or complete loss of Lzts1 downregulates Cdc25C and inhibits Cdk1 activity during mitosis, leading to premature transition from metaphase to anaphase. PMID- 17419987 TI - Actin on multiple fronts to generate a muscle fiber. AB - How the actin cytoskeleton is harnessed to fulfill its diverse cellular functions is a recurrent and intriguing question. In this issue of Developmental Cell, Kim et al. and Massarwa et al. describe a new role for F-actin, and more specifically the actin regulator WASp, in myoblast fusion in Drosophila. PMID- 17419988 TI - Niemann-Pick C proteins in sterol transport and absorption: flies in the ointment. AB - The Niemann-Pick C proteins have slowly emerged as regulators of subcellular lipid transport and sterol absorption at the small intestine. A recent article in Cell Metabolism suggests that in addition to their significant structural and sequence homology, these proteins may orchestrate their functions in a previously unappreciated fashion (Voght et al., 2007). PMID- 17419989 TI - Reactive oxygen species regulate autophagy through redox-sensitive proteases. AB - Starvation induces autophagy through a signal transduction pathway that is not fully understood. In a recent issue of The EMBO Journal, Scherz-Shouval and colleagues (Scherz-Shouval et al., 2007) show that reactive oxygen species (ROS) occurring during starvation serve as signaling molecules that initiate autophagy. PMID- 17419990 TI - The two TORCs and Akt. AB - The regulatory circuits that control the activities of the two distinct target of rapamycin (TOR) complexes, TORC1 and TORC2, and of Akt have been a focus of intense research in recent years. It has become increasingly evident that these regulatory circuits control some of the most fundamental aspects of metabolism, cell growth, proliferation, survival, and differentiation at both the cellular and organismal levels. As such, they also play a pivotal role in the genesis of diseases including cancer, diabetes, aging, and degenerative diseases. This review highlights recent developments aimed at deciphering the interplay between Akt and mTORCs as well as their role in embryonic development and in cancer. PMID- 17419991 TI - A microtubule-independent role for centrosomes and aurora a in nuclear envelope breakdown. AB - Aurora A kinase localizes to centrosomes and is required for centrosome maturation and spindle assembly. Here we describe a microtubule-independent role for Aurora A and centrosomes in nuclear envelope breakdown (NEBD) during the first mitotic division of the C. elegans embryo. Aurora A depletion does not alter the onset or kinetics of chromosome condensation, but dramatically lengthens the interval between the completion of condensation and NEBD. Inhibiting centrosome assembly by other means also lengthens this interval, albeit to a lesser extent than Aurora A depletion. By contrast, centrosomally nucleated microtubules and the nuclear envelope-associated motor dynein are not required for timely NEBD. These results indicate that mitotic centrosomes generate a diffusible factor, which we propose is activated Aurora A, that promotes NEBD. A positive feedback loop, in which an Aurora A-dependent increase in centrosome size promotes Aurora A activation, may temporally couple centrosome maturation to NEBD during mitotic entry. PMID- 17419992 TI - Centrosomes promote timely mitotic entry in C. elegans embryos. AB - Several mitotic regulators, including Cyclin B1/Cdk1, are present at centrosomes prior to mitosis onset, but it is unclear whether centrosomes promote mitotic entry in vivo. Here we developed a sensitive assay in C. elegans embryos for the temporal analysis of mitotic entry, in which the male and female pronuclei undergo asynchronous entry into mitosis when separated from one another. Using this assay, we found that centrosome integrity is necessary for timing mitotic entry. Centrosomes do not function in this instance through their ability to nucleate microtubules. Instead, centrosomes serve to focus the Aurora A kinase AIR-1, which is essential for timely mitotic entry. Furthermore, analysis of embryos in which centrosomes and pronuclei are detached from one another demonstrates that centrosomes are sufficient to promote mitosis onset. Together, our findings support a model in which centrosomes serve as integrative centers for mitotic regulators and thus trigger mitotic entry in a timely fashion. PMID- 17419993 TI - Stimulation of endocytosis and actin dynamics by Oskar polarizes the Drosophila oocyte. AB - In Drosophila, localized activity of oskar at the posterior pole of the oocyte induces germline and abdomen formation in the embryo. Oskar has two isoforms, a short isoform encoding the patterning determinant and a long isoform of unknown function. Here, we show by immuno-electron microscopy that the two Oskar isoforms have different subcellular localizations in the oocyte: Short Oskar mainly localizes to polar granules, and Long Oskar is specifically associated with endocytic membranes along the posterior cortex. Our cell biological and genetic analyses reveal that Oskar stimulates endocytosis, and that its two isoforms are required to regulate this process. Furthermore, we describe long F-actin projections at the oocyte posterior pole that are induced by and intermingled with Oskar protein. We propose that Oskar maintains its localization at the posterior pole through dual functions in regulating endocytosis and F-actin dynamics. PMID- 17419994 TI - WIP/WASp-based actin-polymerization machinery is essential for myoblast fusion in Drosophila. AB - Formation of syncytial muscle fibers involves repeated rounds of cell fusion between growing myotubes and neighboring myoblasts. We have established that Wsp, the Drosophila homolog of the WASp family of microfilament nucleation-promoting factors, is an essential facilitator of myoblast fusion in Drosophila embryos. D WIP, a homolog of the conserved Verprolin/WASp Interacting Protein family of WASp binding proteins, performs a key mediating role in this context. D-WIP, which is expressed specifically in myoblasts, associates with both the WASp-Arp2/3 system and with the myoblast adhesion molecules Dumbfounded and Sticks and Stones, thereby recruiting the actin-polymerization machinery to sites of myoblast attachment and fusion. Our analysis demonstrates that this recruitment is normally required late in the fusion process, for enlargement of nascent fusion pores and breakdown of the apposed cell membranes. These observations identify cellular and developmental roles for the WASp-Arp2/3 pathway, and provide a link between force-generating actin polymerization and cell fusion. PMID- 17419995 TI - A critical function for the actin cytoskeleton in targeted exocytosis of prefusion vesicles during myoblast fusion. AB - Myoblast fusion is an essential step during muscle differentiation. Previous studies in Drosophila have revealed a signaling pathway that relays the fusion signal from the plasma membrane to the actin cytoskeleton. However, the function for the actin cytoskeleton in myoblast fusion remains unclear. Here we describe the characterization of solitary (sltr), a component of the myoblast fusion signaling cascade. sltr encodes the Drosophila ortholog of the mammalian WASP interacting protein. Sltr is recruited to sites of fusion by the fusion-competent cell-specific receptor Sns and acts as a positive regulator for actin polymerization at these sites. Electron microscopy analysis suggests that formation of F-actin-enriched foci at sites of fusion is involved in the proper targeting and coating of prefusion vesicles. These studies reveal a surprising cell-type specificity of Sltr-mediated actin polymerization in myoblast fusion, and demonstrate that targeted exocytosis of prefusion vesicles is a critical step prior to plasma membrane fusion. PMID- 17419996 TI - Liprinalpha1 degradation by calcium/calmodulin-dependent protein kinase II regulates LAR receptor tyrosine phosphatase distribution and dendrite development. AB - Neural activity regulates dendrite and synapse development, but the underlying molecular mechanisms are unclear. Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is an important sensor of synaptic activity, and the scaffold protein liprinalpha1 is involved in pre- and postsynaptic maturation. Here we show that synaptic activity can suppress liprinalpha1 protein level by two pathways: CaMKII mediated degradation and the ubiquitin-proteasome system. In hippocampal neurons, liprinalpha1 mutants that are immune to CaMKII degradation impair dendrite arborization, reduce spine and synapse number, and inhibit dendritic targeting of receptor tyrosine phosphatase LAR, which is important for dendrite development. Thus, regulated degradation of liprinalpha1 is important for proper LAR receptor distribution, and could provide a mechanism for localized control of dendrite and synapse morphogenesis by activity and CaMKII. PMID- 17419997 TI - PLA2 and PI3K/PTEN pathways act in parallel to mediate chemotaxis. AB - Directed cell migration involves signaling events that lead to local accumulation of PI(3,4,5)P(3), but additional pathways act in parallel. A genetic screen in Dictyostelium discoideum to identify redundant pathways revealed a gene with homology to patatin-like phospholipase A(2). Loss of this gene did not alter PI(3,4,5)P(3) regulation, but chemotaxis became sensitive to reductions in PI3K activity. Likewise, cells deficient in PI3K activity were more sensitive to inhibition of PLA(2) activity. Deletion of the PLA(2) homolog and two PI3Ks caused a strong defect in chemotaxis and a reduction in receptor-mediated actin polymerization. In wild-type cells, chemoattractants stimulated a rapid burst in an arachidonic acid derivative. This response was absent in cells lacking the PLA(2) homolog, and exogenous arachidonic acid reduced their dependence on PI3K signaling. We propose that PLA(2) and PI3K signaling act in concert to mediate chemotaxis, and metabolites of PLA(2) may be important mediators of the response. PMID- 17419998 TI - Mek1/2 MAPK kinases are essential for Mammalian development, homeostasis, and Raf induced hyperplasia. AB - The p42/p44 mitogen-activated protein kinase (MAPK) cascade includes Ras, Raf, Mek, and Erk MAPK. To determine the effect of a full knockout at a single level of this signaling pathway in mammals, and to investigate functional redundancy between Mek1 and Mek2, we disrupted these genes in murine and human epidermis. Loss of either protein alone produced no phenotype, whereas combined Mek1/2 deletion in development or adulthood abolished Erk1/2 phosphorylation and led to hypoproliferation, apoptosis, skin barrier defects, and death. Conversely, a single copy of either allele was sufficient for normal development. Combined Mek1/2 loss also abolished Raf-induced hyperproliferation. Human tissue deficient in either Mek isoform was normal, whereas loss of both proteins led to hypoplasia, which was rescued by active Erk2 expression. These data indicate that Mek1/2 are functionally redundant in the epidermis, where they act as a linear relay in the MAPK pathway to mediate development and homeostasis. PMID- 17419999 TI - A double-assurance mechanism controls cell cycle exit upon terminal differentiation in Drosophila. AB - Terminal differentiation is often coupled with permanent exit from the cell cycle, yet it is unclear how cell proliferation is blocked in differentiated tissues. We examined the process of cell cycle exit in Drosophila wings and eyes and discovered that cell cycle exit can be prevented or even reversed in terminally differentiating cells by the simultaneous activation of E2F1 and either Cyclin E/Cdk2 or Cyclin D/Cdk4. Enforcing both E2F and Cyclin/Cdk activities is required to bypass exit because feedback between E2F and Cyclin E/Cdk2 is inhibited after cells differentiate, ensuring that cell cycle exit is robust. In some differentiating cell types (e.g., neurons), known inhibitors including the retinoblastoma homolog Rbf and the p27 homolog Dacapo contribute to parallel repression of E2F and Cyclin E/Cdk2. In other cell types, however (e.g., wing epithelial cells), unknown mechanisms inhibit E2F and Cyclin/Cdk activity in parallel to enforce permanent cell cycle exit upon terminal differentiation. PMID- 17420000 TI - The transcription factor MEF2C is required for craniofacial development. AB - MEF2 transcription factors are well-established regulators of muscle development. We have discovered an unanticipated role for MEF2C in the neural crest, where tissue-specific inactivation results in neonatal lethality due to severe craniofacial defects. We show that MEF2C is required for expression of the Dlx5, Dlx6, and Hand2 transcription factor genes in the branchial arches, and we identify a branchial arch-specific enhancer in the Dlx5/6 locus, which is activated synergistically by MEF2C and Dlx5, demonstrating that these factors interact to induce transcription. Mef2c and Dlx5/6 also interact genetically. Mice heterozygous for either Dlx5/6 or Mef2c are normal at birth and survive to weaning. By contrast, heterozygosity for both Mef2c and Dlx5/6 results in defective palate development and neonatal lethality. Taken together, the studies presented here define a feed-forward transcriptional circuit between the MADS-box transcription factor MEF2C and the homeodomain transcription factors Dlx5 and Dlx6 in craniofacial development. PMID- 17420001 TI - Membrane hemifusion is a stable intermediate of exocytosis. AB - Membrane fusion during exocytosis requires that two initially distinct bilayers pass through a hemifused intermediate in which the proximal monolayers are shared. Passage through this intermediate is an essential step in the process of secretion, but is difficult to observe directly in vivo. Here we study membrane fusion in the sea urchin egg, in which thousands of homogeneous cortical granules are associated with the plasma membrane prior to fertilization. Using fluorescence redistribution after photobleaching, we find that these granules are stably hemifused to the plasma membrane, sharing a cytoplasmic-facing monolayer. Furthermore, we find that the proteins implicated in the fusion process-the vesicle-associated proteins VAMP/synaptobrevin, synaptotagmin, and Rab3-are each immobile within the granule membrane. Thus, these secretory granules are tethered to their target plasma membrane by a static, catalytic fusion complex that maintains a hemifused membrane intermediate. PMID- 17420002 TI - Is location cueing inherently superior to color cueing? Not if color is presented early enough. AB - A possible source for the advantage of location cueing over non-spatial cueing is that orienting attention by a location cue is feasible prior to stimulus onset, whereas that is normally not the case with orienting by a non-spatial cue. To examine how critical that source is for observing an advantage, we eliminated it: In a color-preview condition, subjects were to detect a target presented on the background of one of two differently colored circles (where color-location assignment was random). In a no-preview condition, the circles were both gray, but the target was either red or green (where color assignment was random). Cue type (location vs color) was also manipulated. The color preview in Experiment 1 (in which color onset preceded cue onset) was found helpful: Whereas a substantial disparity in validity effects of the two cue types was obtained with no preview, no significant difference was found when a color preview was introduced. The validity effects of both cue types were found to be about the same also in Experiment 2, in which color onset was exactly synchronized with cue onset, and SOA was manipulated. Furthermore, the absence of an SOA x cue type interaction indicated that the time course of the color cue validity did not lag after the time course of the location cue validity, which seems incompatible with the hypothesis that a color cue cannot affect orienting without first computing a location from it prior to cue onset. Overall, the results suggest that the time course of color cueing is not inherently different from that of location cueing once its main disadvantages are removed. PMID- 17420003 TI - A DNA electrochemical sensor based on nanogold-modified poly-2,6 pyridinedicarboxylic acid film and detection of PAT gene fragment. AB - A new DNA electrochemical biosensor is described for electrochemical impedance spectroscopy (EIS) detection of the sequence-specific DNA related to PAT transgene in the transgenic plants. Poly-2,6-pyridinedicarboxylic acid film (PDC) was fabricated by electropolymerizing 2,6-pyridinedicarboxylic acid on the glassy carbon electrode (GCE). The gold nanoparticles (NG) were modified on the PDC/GCE to prepare NG/PDC/GCE, and then DNA probe (ssDNA) was immobilized on the NG/PDC/GCE by the interaction of NG with DNA. The immobilization of NG and the immobilization and hybridization of DNA probe were characterized with differential pulse voltammetry (DPV) and cyclic voltammetry (CV) using methylene blue (MB) as indicator and EIS. MB had a couple of well-defined CV peaks at the NG/PDC/GCE, and these redox peak currents increased after the immobilization of the DNA probe. After the hybridization of the DNA probe with the complementary single-stranded DNA (cDNA), the redox peak currents of MB decreased greatly. The electron transfer resistance (R(et)) of the electrode surface in EIS in [Fe(CN)(6)](3-/4-) solution increased after the immobilization of the DNA probe on the NG/PDC/GCE. The hybridization of the DNA probe with cDNA made R(et) increase further. EIS was used for the label-free detection of the target DNA. The NG modified on the PDC dramatically enhanced the immobilization amount of the DNA probe and greatly improved the sensitivity of DNA detection. The difference between the R(et) value at the ssDNA/NG/PDC/GCE and that at hybridization DNA modified electrode (dsDNA/NG/PDC/GCE) was used as the signal for detecting the PAT gene fragment with the dynamic range from 1.0x10(-10) to 1.0x10(-5)mol/L. A detection limit of 2.4x10(-11)mol/L could be estimated. PMID- 17420004 TI - Molecular cloning and functional characterization of a novel decarboxylase from uncultured microorganisms. AB - The metagenomic library approach has been used successfully to isolate novel biocatalyst genes from uncultured microorganisms. We report the cloning of a novel decarboxylase gene by sequence-based screening of a plasmid metagenomic library constructed with DNA from alkaline polluted soils. The gene was named undec1 A and had an open reading frame of 1077 base pairs. It encoded a 359 amino acid polypeptide with a molecular mass of 38 kDa. The predicted protein had 58% similarity to a decarboxylase from Chlorobium phaeobacteroides BS1. The putative decarboxylase gene was subcloned into pETBlue-2 vector and overexpressed in Escherichia coli Tuner (DE3) pLac. The recombinant protein was purified to homogeneity. Functional characterization with liquid chromatography-mass spectrometry confirmed that the recombinant Undec1 A protein catalyzed the decarboxylation of L-cysteine to form cysteamine. PMID- 17420005 TI - Enhanced IL-1beta-induced IL-8 production in cystic fibrosis lung epithelial cells is dependent of both mitogen-activated protein kinases and NF-kappaB signaling. AB - Transcription nuclear factor-kappaB (NF-kappaB) is hyperactivated in cystic fibrosis (CF) lung epithelial cells, and participates in exaggerated IL-8 production in the CF lung. We recently found that rapid activation of NF-kappaB occurred in a CF lung epithelial IB3-1 cell line (CF cells) upon IL-1beta stimulation, which was not observed in its CFTR-corrected lung epithelial S9 cell line (corrected cells). To test whether other signaling pathways such as that of mitogen-activated protein kinases (MAPKs) could be involved in IL-1beta-induced IL-8 production of CF cells, we investigated ERK1/2, JNK, and p38MAP signaling compared to NF-kappaB. Within 30min, exposure to IL-1beta caused high activation of NF-kappaB, ERK1/2, p38MAP but not JNK in CF cells compared to corrected cells. Treatment of IL-1beta-stimulated CF cells with a series of chemical inhibitors of NF-kappaB, ERK1/2, and p38MAP, when used separately, reduced slightly IL-8 production. However, when used together, these inhibitors caused a blockade in IL 1beta-induced IL-8 production in CF cells. Understanding of the cross-talk between NF-kappaB and MAPKs signaling in CF lung epithelial cells may help in developing new therapeutics to reduce lung inflammation in patients with CF. PMID- 17420006 TI - Epigenetic regulation of clusterin/apolipoprotein J expression in retinal pigment epithelial cells. AB - Age-related macular degeneration (AMD) is the leading cause of blindness worldwide. AMD is characterized by the deposition of drusen aggregates under the retinal pigment epithelium (RPE). Clusterin/apo J, a multifunctional secreted chaperone, is one of the major proteins accumulating in drusen deposits. The regulation of clusterin expression is not well characterized but the promoter of clusterin contains a CpG-rich methylation domain. Since aging affects both DNA methylation and histone acetylation status, the epigenetic regulation might have an important role in clusterin/apo J expression. Our purpose was to elucidate whether the induction of DNA hypomethylation with 5-aza-2'-deoxycytidine (AZA) and histone hyperacetylation with trichostatin A (TSA) could affect the clusterin transcription, protein levels, and secretion in retinal pigment epithelial cells. We observed that both TSA and AZA treatments induced a prominent increase in the expression levels of clusterin mRNA and protein in ARPE-19 cells, as well as in the secretion of clusterin protein. Furthermore, valproic acid, an antiepileptic drug and a recently identified inhibitor of histone deacetylases (HDAC), induced a significant increase in clusterin protein expression and secretion in retinal pigment epithelial cells. HDAC inhibitors are characterized as inhibitors of angiogenesis, and clusterin as a complement inhibitor. Our results indicate that epigenetic factors regulate the clusterin expression of RPE cells and thus might affect the pathogenesis of AMD via the inhibition of angiogenesis and inflammation. PMID- 17420007 TI - A dynamically changing intracellular water network serves as a universal regulator of the cell: the water-governed cycle. AB - The functioning of enzymes and protein folding are well known to be assisted by the surrounding chaperoning water molecules, which are connected to the protein via non-covalent, dynamically changing chemical bonds. A molecular intracellular network of weak non-covalent connections may be presumed to exist in living cells. The roles of such non-covalent networks are examined in terms of a molecular model which postulates a universal enzyme and biochemical mechanism regulating the maintenance of chemical stability in living cells. PMID- 17420008 TI - Synthesis and conformational analysis of an alpha-D-mannopyranosyl-(1-->2)-alpha D-mannopyranosyl-(1-->6)-alpha-D-mannopyranose mimic. AB - A mimic of a (1-->2),(1-->6)-mannotrioside was synthesized by replacing the central mannose unit with an enantiomerically pure, conformationally stable trans diaxial cyclohexanediol. The three-dimensional structure of the molecule was investigated by NMR spectroscopy supported by molecular modelling and was compared to the known features of the natural mannotrioside. PMID- 17420009 TI - A new 5-deoxyflavone glycoside from the aerial parts of Calea clausseniana. AB - A new 5-deoxyflavone glycoside, identified as 7-O-(alpha-L-rhamnopyranosyl-(1- >2)-beta-D-glucopyranosyl)-3',4',7-trihydroxyflavone (1), was isolated from the aerial parts of Calea clausseniana. Its structure was determined by spectral analysis. PMID- 17420010 TI - Augmentation of innate immunity by low-dose irradiation. AB - The effect of low-dose irradiation on the immune system was investigated in mice. When a 0.2 Gy dose of X-ray irradiation was administered every other day for a total of four times, the number of lymphocytes yielded by the liver, spleen and thymus decreased at the initial stage (around day 10). At this stage, NK cells, extrathymic T cells and NKT cells were found to be radioresistant. In other words, conventional lymphocytes were radiosensitive, even in the case of low-dose irradiation. However, the number of lymphocytes in all tested immune organs increased beyond the control level at the recovery stage (around day 28). Enumeration of the absolute number of lymphocyte subsets showed that the most prominently expanding populations were NK cells, extrathymic T cells and NKT cells, especially in the liver where primordial lymphocytes are primarily present. Functional and phenotypic activation of these populations also occurred at the recovery stage. It raised a possibility that an initial activation of macrophages by low-dose irradiation then mediated the present phenomenon. These results suggest that low-dose irradiation eventually has the potential to induce a hormesis effect on the immune system. PMID- 17420011 TI - Penta-acylated lipopolisaccharide binds to murine MD-2 but does not induce the oligomerization of TLR4 required for signal transduction. AB - A mutant lipopolysaccharide (LPS) lacking a myristate chain in lipid A was shown to be non-pathogenic both in humans and mice. The mutant penta-acylated LPS from the lpxM-strain did not induce TNF-alpha production in murine peritoneal macrophages, or activation of NF-kappaB in transfected cells expressing murine TLR4/MD-2. We prepared a recombinant murine MD-2 in Escherichia coli (E. coli), and examined the binding function. Unexpectedly, specific binding was detected to both wild type and mutant LPS. However, the mutant LPS did not induce conformation changes or oligomerization of TLR4, which have been shown to be required for signal transduction. Mutant LPS appears to fail to induce appropriate conformational changes, resulting in oligomerization of the murine complex for triggering cell responses. PMID- 17420012 TI - Histamine H1 and H2 receptors but not H4 receptors are upregulated during bone marrow regeneration. AB - The role of histamine receptors in radiation-induced bone marrow (BM) regeneration was investigated with aspects of functional genomics. H1R and H2R mRNA expression increased during regeneration in both histidine decarboxylase knockout (HDC-/-) and wild type (HDC+/+) mice, though to a lesser extent in HDC-/ mice. H4R mRNA expression was downregulated in both groups. Mainly CD34+ cells were responsible for the elevation of intracellular histamine and HDC content in HDC+/+ BM cell populations. The differential changes in the expression of its receptors, and also its elevated levels in hematopoietic progenitors support the regulatory role of histamine in BM regeneration, that could be further explored by future gene expression studies. PMID- 17420013 TI - A quinoxaline 1,4-di-N-oxide derivative induces DNA oxidative damage not attenuated by vitamin C and E treatment. AB - Some anticancer compounds are pro-drugs which give rise to toxic species through enzymatic reduction. The quinoxaline-di-N-oxide derivative Q-85 HCl (7-chloro-3 [[(N,N-dimethylamino)propyl]amino]-2-quinoxalinecarbonitrile 1,4-di-N-oxide hydrochloride) is a bioreductive compound selectively toxic in hypoxia. Due to the possibility of secondary tumors the study of the genotoxic capability of antitumoral drugs is very important. The aim of this study was to assess the ability of Q-85 HCl to produce reactive oxygen species (ROS) and oxidative DNA damage in Caco-2 cells, both in hypoxia and in well-oxygenated conditions. Secondly, we attempted to evaluate the effect of vitamins C and E under hypoxic and normoxic conditions, in order to determine if these antioxidant substances modify Q-85 HCl effect in hypoxic cells or possibly exert a protective action in normal cells. Caco-2 cells were treated with Q-85 HCl for 2h, at high concentrations in normoxia (0.1-5 microM) and at low concentrations in hypoxia (0.002-0.1 microM). In normoxia, a dose-related significant increase in intracellular ROS level was evident; in hypoxia all the concentrations produced very high level of ROS. Just after the treatment and 24h later, oxidative DNA damage was evaluated by the modified comet assay after post-digestion of the cells with formamidopyrimidine-DNA glycosylase (FPG) and endonuclease III (Endo III). Q-85 HCl treatment evoked a significant dose-dependent increase in the total comet score of the cells both in hypoxia and normoxia, indicating that this compound or some metabolite is able to oxidize purine and pyrimidine bases. After 24h DNA damage caused by the compound was completely repaired with only one exception: cells treated with the highest concentration of Q-85 HCl in hypoxia and post-digested with FPG. Vitamin C (5-100 microM) and vitamin E (500-400 microM) did not have a pro-oxidant effect in Caco-2 cells. Treatment of cells with vitamin C (10 microM) or vitamin E (100 microM) did not significantly reduce oxidative DNA damage in hypoxia and normoxia. In conclusion, the use of these vitamins would not hinder toxicity against hypoxic cells, but a protective effect in normoxic cells was not evident. PMID- 17420014 TI - Nippostrongylus brasiliensis: identification of intelectin-1 and -2 as Stat6 dependent genes expressed in lung and intestine during infection. AB - Elimination of the helminth parasite Nippostrongylus brasiliensis from infected mice is mediated by IL-4 or IL-13 and dependent on the IL-4Ralpha chain and the transcription factor Stat6 in non-hematopoietic cells. However, it is not clear which Stat6-dependent effector molecules mediate worm expulsion. We identified intelectin-1 and -2 as Stat6-dependent genes that are induced during infection. Intelectins can bind galactofuranose, a sugar present only in microorganisms and might therefore serve as microbial pattern element. To analyze whether constitutive expression of intelectin-1 or -2 leads to accelerated pathogen clearance, transgenic mice were generated which express high levels of these genes selectively in the lung. Infection with N. brasiliensis or Mycobacterium tuberculosis did not result in accelerated pathogen clearance in transgenic as compared to wild-type mice. Further, no significant modulation of the immune response in lung or lymph nodes was observed. Thus, under these conditions, intelectins did not enhance pathogen clearance. PMID- 17420015 TI - Cysteine proteinase 30 in clinical isolates of T. vaginalis from symptomatic and asymptomatic infected women. AB - A cysteine proteinase of 30 kDa (CP30) of Trichomonas vaginalis, is known to play a role in cytoadherence of the parasite to host cells. However, the CP30 activity in clinical isolates from symptomatic and asymptomatic patients has not been analyzed. In the present study, CP30 was detected in 20 fresh and long-term culture maintained T. vaginalis isolates each from symptomatic and asymptomatic women by substrate gel electrophoresis and immunoblotting. Though CP30 was detected in all the fresh isolates from 20 symptomatic and 20 asymptomatic women, the intensity of CP30 band was significantly higher in isolates from symptomatic as compared to asymptomatic women indicating higher expression in former. CP30 was found in all the 20 long-term cultured isolates from symptomatic whereas only in 70% of asymptomatic women indicating that CP30 expression is a more stable characteristic of symptomatic isolates. The isolates from symptomatic women, demonstrated significantly higher cytoadherence to VECs as compared to asymptomatic women. In both the types of isolates, this cytoadherence was inhibited significantly by CP30 specific hyperimmune serum. These results confirm that CP30 is an important virulence factor of T. vaginalis and has an important role in cytoadherence to VECs and thus has a role in pathogenesis of trichomoniasis. PMID- 17420016 TI - Schistosoma mansoni: Heterologous complementation of a yeast null mutant by SmRbx, a protein similar to a RING box protein involved in ubiquitination. AB - The SCF (Skp1-Cul1-F-box) complex is one of the several E3 ligase enzymes and it catalyzes protein ubiquitination and degradation by the 26S proteasome. Rbx1 is a member of the SCF complex in humans and HRT1 is its yeast orthologue. A cDNA encoding a Schistosoma mansoni Rbx1 homolog was cloned and functionally characterized. Heterologous functional complementation in yeast showed that the worm SmRbx gene was able to complement the HRT1yeast null mutation. Gene deletion constructs for N- and C-termini truncated proteins were used to transform hrt1(-) yeast mutant strains, allowing us to observe that regions reported to be involved in the interaction with cullin1 (Cul1) were essential for SmRbx function. Yeast two-hybrid assays using SmRbx and yeast Cul1 confirmed that SmRbx, but not the mutant SmRbxDelta24N, lacking the N-terminus of the protein, was capable of interacting with Cul1. These results suggest that SmRbx protein is involved in the SCF complex formation. PMID- 17420017 TI - Molecular characterization and seasonal changes in gonadal expression of a thyrotropin receptor in the European sea bass. AB - The thyroid stimulating hormone (TSH) is a glycoprotein synthesized and secreted from thyrotrophs of the anterior pituitary gland. It acts by binding to and activating its specific receptor, the TSHR, to induce the synthesis and secretion of thyroid hormones. Recent studies conducted in diverse fish species suggest a direct role of TSH on gonadal physiology. In this work, we describe the cloning of a cDNA encoding a TSHR which was isolated from the gonads of the European sea bass (Dicentrarchus labrax). The mature protein displays typical features of the members of the glycoprotein hormone receptor family and shows the highest amino acid sequence identity with the TSHRs of other fish species. An insertion of approximately 50 amino acids, specific for the TSHR subfamily is also present in the carboxyl end of the extracellular domain of the sbsTSHR. By RT-PCR analysis, we demonstrate the extrathyroidal expression of sbsTSHR in numerous tissues of the sea bass. Also, two transcripts that differ in the length of their 3' untranslated regions were found. They reflect the use of alternative polyadenylation cleavage sites. Seasonal changes in sbsTSHR mRNA levels in female and male sea bass during the first ovarian and testicular recrudescence suggest that in females the TSHR could participate in active vitellogenesis and in the regulation of gamete maturation and ovulation, whereas in males, the TSHR would be involved in the regulation of processes that occur during the early stages of the gonadal development and also of gamete maturation and spermiation. The results of this work indicate that a sbsTSHR has been cloned from the testis of the European sea bass and they provide the basis for future studies concerning the function of TSHR in this species. PMID- 17420018 TI - Mass spectrometric identification of pEGFYSQRYamide: a crustacean peptide hormone possessing a vertebrate neuropeptide Y (NPY)-like carboxy-terminus. AB - In invertebrates, peptides possessing the carboxy (C)-terminal motif -RXRFamide have been proposed as the homologs of vertebrate neuropeptide Y (NPY). Using matrix assisted laser desorption/ionization mass spectrometry, in combination with sustained off-resonance irradiation collision-induced dissociation and chemical and enzymatic reactions, we have identified the peptide pEGFYSQRYamide from the neuroendocrine pericardial organ (PO) of the crab Pugettia producta. This peptide is likely the same as that previously reported, but misidentified, as PAFYSQRYamide in several earlier reports (e.g. [Li, L., Kelley, W.P., Billimoria, C.P., Christie, A.E., Pulver, S.R., Sweedler, J.V., Marder, E. 2003. Mass spectrometric investigation of the neuropeptide complement and release in the pericardial organs of the crab, Cancer borealis. J. Neurochem. 87, 642-656; Fu, Q., Kutz, K.K., Schmidt, J.J., Hsu, Y.W., Messinger, D.I., Cain, S.D., de la Iglesia, H.O., Christie, A.E., Li, L. 2005. Hormone complement of the Cancer productus sinus gland and pericardial organ: an anatomical and mass spectrometric investigation. J. Comp. Neurol. 493, 607-626.]). The -QRYamide motif contained in pEGFYSQRYamide is identical to that present in many vertebrate members of the NPY superfamily. Mass spectrometric analysis conducted on the POs of several other decapods showed that pEGFYSQRYamide is present in three other brachyurans (Cancer borealis, Cancer irroratus and Cancer productus) as well as in one species from another decapod infraorder (Lithodes maja, an anomuran). Thus, our findings show that at least some invertebrates possess NPY-like peptides in addition to those exhibiting an -RXRFamide C-terminus, and raise the question as to whether the invertebrate -QRYamides are functionally and/or evolutionarily related to the NPY superfamily. PMID- 17420019 TI - Specific P-selectin and P-selectin glycoprotein ligand-1 genotypes/haplotypes are associated with risk of incident CHD and ischemic stroke: the Atherosclerosis Risk in Communities (ARIC) study. AB - OBJECTIVE: P-selectin (PSEL) and its ligand, P-selectin glycoprotein ligand-1 (PSGL-1), play key roles in both the inflammatory response and the atherosclerotic process, but there are conflicting results regarding the affect of PSEL and PSGL-1 gene variation on risk for cardiovascular and cerebrovascular disease. We tested the association of four PSEL and two PSGL-1 polymorphisms with incident coronary heart disease (CHD) and ischemic stroke among 13,875 participants in the prospective Atherosclerosis Risk in Communities (ARIC) study. We also tested common haplotypes in the PSEL and PSGL-1 genes to assess associations with incident CHD and ischemic stroke. METHODS AND RESULTS: Incident ischemic stroke and CHD were identified through annual telephone calls and hospital and death certificate surveillance. Five hundred and twenty-five validated ischemic stroke and 1654 CHD events were identified. Allele frequencies for all PSEL and PSGL-1 polymorphisms were markedly different between whites and African Americans; therefore, all analyses were performed race-specific. Independent analyses showed the PSEL 290NN genotype to be a significant predictor of CHD in whites (HRR 1.30, 95%CI 1.00-1.70, P=0.05). PSGL-1 genotypes carrying the 62I allele were significantly protective for incident CHD (HRR 0.53, 95%CI 0.31-0.92, P=0.02) and ischemic stroke (HRR 0.73, 95%CI 0.55-0.97, P=0.03) in African Americans. Haplotype analyses showed the PSEL NNVP haplotype to be a significant predictor of incident CHD in whites (HRR 2.09, 95%CI 1.23-3.55, P=0.006). No significant haplotype findings were observed in African Americans. CONCLUSIONS: PSEL S290N, in single polymorphism analysis and in the haplotypic background with T715P, was associated with increased risk of incident CHD in whites. The PSGL-1 M62I polymorphism was associated with decreased risk of both incident CHD and stroke in African Americans. These findings illustrate the complex relationship between genetic variation and disease in different racial groups. PMID- 17420020 TI - Resialylation of sialic acid deficit vascular endothelium, circulating cells and macromolecules may counteract the development of atherosclerosis: a hypothesis. AB - Deficit sialylation of vascular endothelium, circulating cells and macromolecules has been associated with the development of atherosclerosis. On the other hand, an elevated serum level of sialic acid is a long-lasting marker of atherosclerosis and complications from atherosclerosis. One may speculate that the inverse associations with atherosclerosis risk are due to some common underlying cause. One mission for the elevated serum sialic acid level might be to act as a substrate for resialylation of sialic acid deficit structures and thereby counteract the atherosclerotic process. PMID- 17420021 TI - Multivariate calibration. AB - The bases of multivariate calibration are presented with special attention to some points usually not considered or underevaluated, i.e., the sampling design, the number of samples necessary to obtain a reliable regression model, the effect of noisy predictors, the significance of the parameters used to evaluate the performance ability of the regression model. PMID- 17420022 TI - Determination of perfluorinated compounds in wastewater and river water samples by mixed hemimicelle-based solid-phase extraction before liquid chromatography electrospray tandem mass spectrometry detection. AB - A comparative study on the use of cetyltrimethylammonium bromide (CTAB)-coated silica and sodium dodecyl sulphate (SDS)-coated alumina mixed hemimicelles-based solid-phase extraction (SPE) for the pre-concentration of six perfluorinated compounds (PFCs) in environmental water samples was presented. The six analytes heptafluorobutyric acid (HFBA), perfluoroheptanic acid (PFHeA), perfluorooctanic acid (PFOA), perfluorooctanic sulfonic (PFOS), perfluorononanic acid (PFNA) and perfluorodecanic acid (PFDeA) were quantitatively retained on both sorbent materials. The cationic surfactant (CTAB adsorbed onto silica) was more appropriate for SPE of PFCs. The main factors affecting adsolubilization of PFCs including the amount of surfactant, pH of solution, sample loading volume and desorption were investigated and optimized. Concentration factor of 500 were achieved by SPE of 500 mL of several environmental water samples. The method detection limits obtained for HFBA, PFHeA, PFOA, PFOS, PFNA and PFDeA were 0.10, 0.28, 0.07, 0.20, 0.10 and 0.05 ng/L, respectively. The relative standard deviation of recoveries ranged from 2 to 8%, which indicated good method precision. PMID- 17420023 TI - Rapid determination of total trihalomethanes index in drinking water. AB - A method for the rapid determination of total trihalomethanes (THMs) index in drinking water has been developed by using a headspace-mass spectrometry (HS-MS) system and partial least squares (PLS) multivariate regression approach. Due to the presence of residual amounts of chlorine and organic matter in the drinking water, the use of a quenching reagent in order to avoid THM generation during the sample manipulation is necessary. The optimization experiments revealed that ascorbic acid was the best quenching reagent compared with sodium thiosulfate and ammonium sulfate. The use of a classification chemometric technique as soft independent modeling of class analogy before the PLS regression improved the results obtained in the prediction of the total THMs index, lowering the relative standard error of prediction (RSEP) from 11.4% to lower than 6.0%. The results obtained by the proposed HS-MS method were compared with those provided by a conventional chromatographic method after analyzing 20 real drinking water samples. A good agreement in the results was observed and no systematic differences were found, which corroborates the good performance of the proposed method. PMID- 17420024 TI - Viscous fingering in packed chromatographic columns: linear stability analysis. AB - When a fluid is displaced by a less viscous one in a porous medium, a hydrodynamic instability appears leading to the formation of some kind of fingers of the upstream fluid invading the downstream one, hence the name "viscous fingering" (VF) given to this instability. In a LC column, such an instability is likely to appear at that of the two interfaces between the sample and the eluent which exhibits an unfavorable viscosity contrast. It leads to distorted peak shapes and contributes to peak broadening. This phenomenon has been observed for long in SEC and more recently in RPLC on elution peak shapes as well as with various methods of in-column visualization. A simplistic LC column model is described to explain the origin of the VF instability and its characteristics. The general principles for analyzing hydrodynamic instabilities are described and the results of the linear stability analysis performed by Tan and Homsy [C.T. Tan, G.M. Homsy, Phys. Fluids 29 (1986) 3549 [1]], at the onset of the VF phenomenon for a step interface between two fluids are here applied to typical operating conditions encountered in analytical LC. The most probable growth rate and wavelength (linked to the finger width) of the instability are given in terms of particle size and solute diffusion coefficient, with particular emphasis on the role of the carrier velocity. Previously published qualitative observations about VF in chromatography are examined and interpreted at the light of this theory. The role of the column geometry on the development of the instability, the possible sources of noise or fluctuations triggering the instability, and the various experimental situations in which a significant viscosity contrast is encountered in LC are discussed. PMID- 17420025 TI - Using tolerance intervals in pre-study validation of analytical methods to predict in-study results. The fit-for-future-purpose concept. AB - It is recognized that the purpose of validation of analytical methods is to demonstrate that the method is suited for its intended purpose. Validation is not only required by regulatory authorities, but is also a decisive phase before the routine use of the method. For a quantitative analytical method the objective is to quantify the target analytes with a known and suitable accuracy. For that purpose, first, a decision about the validity of the method based on prediction is proposed: a method is declared proper for routine application if it is considered that most of the future results generated will be accurate enough. This can be achieved by using the "beta-expectation tolerance interval" (accuracy profile) as the decision tool to assess the validity of the analytical method. Moreover, the concept of "fit-for-purpose" is also proposed here to select the most relevant response function as calibration curve, i.e. choosing a response function based solely on the predicted results this model will allow to obtain. This paper reports four case studies where the results obtained with quality control samples in routine were compared to predictions made in the validation phase. Predictions made using the "beta-expectation tolerance interval" are shown to be accurate and trustful for decision making. It is therefore suggested that an adequate way to conciliate both the objectives of the analytical method in routine analysis and those of the validation step consists in taking the decision about the validity of the analytical method based on prediction of the future results using the most appropriate response function curve, i.e. the fit-for future-purpose concept. PMID- 17420026 TI - Analysis of recent pharmaceutical regulatory documents on analytical method validation. AB - All analysts face the same situations as method validation is the process of proving that an analytical method is acceptable for its intended purpose. In order to resolve this problem, the analyst refers to regulatory or guidance documents, and therefore the validity of the analytical methods is dependent on the guidance, terminology and methodology, proposed in these documents. It is therefore of prime importance to have clear definitions of the different validation criteria used to assess this validity. It is also necessary to have methodologies in accordance with these definitions and consequently to use statistical methods which are relevant with these definitions, the objective of the validation and the objective of the analytical method. The main purpose of this paper is to outline the inconsistencies between some definitions of the criteria and the experimental procedures proposed to evaluate those criteria in recent documents dedicated to the validation of analytical methods in the pharmaceutical field, together with the risks and problems when trying to cope with contradictory, and sometimes scientifically irrelevant, requirements and definitions. PMID- 17420027 TI - Simultaneous determination of various pharmaceutical compounds in water by solid phase extraction-liquid chromatography-tandem mass spectrometry. AB - A solid-phase extraction (SPE) followed by liquid chromatography-electrospray ionisation tandem mass spectrometry (LC-ESI-MS-MS) method was developed and validated for simultaneous analysis of 11 pharmaceutical residues (propranolol, sulfamethoxazole, meberverine, thioridazine, carbamazepine, tamoxifen, mecoprop, indomethacine, diclofenac, meclofenamic acid and monensin) in environmental water samples. The collision energy chosen for the multiple reaction monitoring (MRM) experiment was optimised. A number of parameters that may affect the recovery of the pharmaceuticals, such as the type of SPE cartridges, eluents, as well as water properties including pH value, salinity and concentration of colloid and surfactant were investigated. It is shown that the Oasis HLB SPE cartridge produced the best recoveries of the target pharmaceuticals while methanol was efficient in eluting pharmaceuticals from SPE cartridges. The recovery of some target compounds was enhanced with increasing salinity, but reduced by increasing pH value, and unaffected by surfactant concentration (0-10 microg/l). The recovery of most compounds was slightly increased by the presence of colloids (0 10 mg/l), which however caused a reduction in recovery for sulfamethoxazole and meberverine. The optimised method was further verified by performing spiking experiments in river water and seawater matrices, with good recovery and reproducibility for all except two compounds. The established method was successfully applied to environmental water samples from East Sussex, UK, for the determination of the target pharmaceuticals. PMID- 17420028 TI - Issues in human auditory development. AB - The human auditory system is often portrayed as precocious in its development. In fact, many aspects of basic auditory processing appear to be adult-like by the middle of the first year of postnatal life. However, processes such as attention and sound source determination take much longer to develop. Immaturity of higher level processes limits the processing of both simple and complex sounds by infants and children. Young listeners with impaired hearing may be at a particular disadvantage, in that they must make sense of sounds on the basis of a degraded representation using immature perceptual strategies. LEARNING OUTCOMES: (1) Readers will be able to describe three stages of human auditory development. (2) Readers will be able to describe how experience with sound is important in auditory development. (3) Readers will be able to describe the role of attention and other higher-level processes in early audition. PMID- 17420029 TI - Natural population dynamics of entomopathogenic nematode Steinernema affine (Steinernematidae) under dry conditions: Possible nematode persistence within host cadavers? AB - The effect of dry conditions on the population dynamics of the entomopathogenic nematode (EPN) Steinernema affine was studied for one month in the exceptionally dry period in the summer of 2003 in the oak wood in the vicinity of Ceske Budejovice, Czech Republic. Soil moisture, soil temperature, and the abundance of suitable insect hosts were monitored. The abundance of infective juveniles (IJs) was correlated with soil moisture and both these values were gradually decreasing during the study period and finally rapidly increased at the end of the investigation. During this period there was a decline in the number of insects suitable as hosts for S. affine, but not in numbers of unsuitable insects. We hypothesise that the observed decrease in IJ numbers was probably caused by the persistence of IJs in host cadavers due to low ambient moisture. PMID- 17420030 TI - In vitro genotoxicity of PAH mixtures and organic extract from urban air particles part II: human cell lines. AB - Principal aims of this study were at first, to find a relevant human derived cell line to investigate the genotoxic potential of PAH-containing complex mixtures and second, to use this cell system for the analysis of DNA adduct forming activity of organic compounds bound onto PM10 particles. Particles were collected by high volume air samplers during summer and winter periods in three European cities (Prague, Kosice, and Sofia), representing different levels of air pollution. The genotoxic potential of extractable organic matter (EOM) was compared with the genotoxic potential of individual carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) as well as their artificial mixtures. Metabolically competent human hepatoma HepG2 cells, confluent cultures of human diploid lung fibroblasts (HEL), and the human monocytic leukemia cell line THP-1 were used as models. DNA adducts were analyzed by (32)P-postlabeling. The total DNA adduct levels induced in HepG2 cells after exposure to EOMs were higher than in HEL cells treated under the same conditions (15-190 versus 2-15adducts/10(8) nucleotides, in HepG2 and HEL cells, respectively). THP-1 cells exhibited the lowest DNA adduct forming activity induced by EOMs (1.5-3.7adducts/10(8) nucleotides). A direct correlation between total DNA adduct levels and c-PAH content in EOM was found for all EOMs in HepG2 cells incubated with 50microg EOM/ml (R=0.88; p=0.0192). This correlation was even slightly stronger when B[a]P content in EOMs and B[a]P-like adduct spots were analyzed (R=0.90; p=0.016). As THP-1 cells possess a limited metabolic capacity for most c-PAHs to form DNA reactive intermediates and are also more susceptible to toxic effects of PAHs and various EOM components, this cell line seemed to be an inappropriate system for genotoxicity studies of PAH-containing complex mixtures. The seasonal variability of genotoxic potential of extracts was stronger than variability among the three localities studied. In HepG2 cells, the highest DNA adduct levels were induced by EOM collected in Prague in the winter period, followed by Sofia and Kosice. However, in the summer sampling period, the order was quite opposite: Kosice>Sofia>Prague. When the EOM content per m(3) of air was taken into consideration in order to compare real exposures of humans to genotoxic compounds in all three localities, extracts from respirable dust particles collected in Sofia exhibited the highest genotoxicity regardless of the sampling period. The results indicate that most of DNA adducts detected in cells incubated with EOMs have their origin in low concentrations of c-PAHs representing 0.03-0.17% of EOM total mass. Finally, our results suggest that HepG2 cells have a metabolic capacity for PAHs similar to human hepatocytes and represent therefore the best in vitro model for investigating the genotoxic potential of complex mixtures containing PAHs among the three cell lines tested in this study. PMID- 17420031 TI - 1,3-Butadiene exposure and cardiovascular disease. AB - This review summarizes the epidemiologic, biochemical and genetic evidence associating occupational, environmental or experimental exposure to 1,3-butadiene (BD) with subsequent development of cardiovascular disease, with the primary focus on atherosclerosis. The potential role of BD in the known atherosclerotic effects of environmental tobacco smoke as well as correlations between polymorphisms in BD phase II enzymes and development of atherosclerosis are presented. PMID- 17420032 TI - Effects of polycyclic aromatic hydrocarbons (PAHs) in environmental pollution on exogenous and oxidative DNA damage (EXPAH project): description of the population under study. AB - The EXPAH project was a molecular epidemiology study whose aims were to evaluate the hypothesis that polycyclic aromatic hydrocarbons (PAHs) are a major source of genotoxic activities of organic mixtures associated with air pollution. Biomarkers of exposure, effects and susceptibility, and oxidative DNA damage were measured in three PAH-exposed populations from Prague (Czech Republic), Kosice (Slovakia) and Sofia (Bulgaria). Control populations were included from each city. In total 356 individuals were enrolled. A questionnaire was used to determine life style/dietary factors. Ambient air exposure was measured by stationary monitoring, and personal exposure monitoring was also carried out. The characteristics of the population are described in this paper together with their personal exposure to carcinogenic PAHs (c-PAHs). The dose of c-PAH exposure was found to vary between the occupationally exposed (e.g. policemen and bus drivers) and the control populations in each country, and also varied from country to country. PMID- 17420033 TI - Role of estrogen receptors in neuroprotection by estradiol against MPTP toxicity. AB - Estradiol protects against striatal dopamine terminal loss caused by the neurotoxin MPTP in mice. This effect of estradiol is thought to be mediated by an interaction with estrogen receptors (ER), of which there are two: ERalpha and ERbeta. In the present study, the role of these two ERs in MPTP toxicity and its neuroprotection by estradiol was investigated using ER knock out mice (ERKO). MPTP (7, 9, or 11 mg/kg administered four times at 2h intervals) caused a dose dependent decrease in striatal dopamine and dopamine metabolite DOPAC concentrations in wild type (WT) mice. The degree of dopamine and DOPAC depletion after MPTP was greater in the ERKOalpha mice than WT mice, whereas the ERKObeta mice exhibited no change in MPTP sensitivity. ERKObeta mice showed a lower DA turnover than WT and ERKOalpha mice. WT, ERKOalpha and ERKObeta mice were also treated for 10 days with exogenous estradiol and on day 5 of treatment were challenged with MPTP (9 mg/kg administered four times at 2h intervals). In the WT mice, estradiol partially prevented the MPTP-induced decrease in striatal dopamine and DOPAC concentrations. However, estradiol treatment was without significant neuroprotective effects in the ERKOalpha and ERKObeta mice. These results show a greater susceptibility to MPTP toxicity of ERKOalpha mice compared to WT and ERKObeta mice and a role for both ER receptors in striatal DA neuroprotection. PMID- 17420034 TI - The calculating hemispheres: studies of a split-brain patient. AB - The purpose of the study was to investigate simple calculation in the two cerebral hemispheres of a split-brain patient. In a series of four experiments, the left hemisphere was superior to the right in simple calculation, confirming the previously reported left hemisphere specialization for calculation. In two different recognition paradigms, right hemisphere performance was at chance for all arithmetic operations, with the exception of subtraction in a two-alternative forced choice paradigm (performance was at chance when the lure differed from the correct answer by a magnitude of 1 but above chance when the magnitude difference was 4). In a recall paradigm, the right hemisphere performed above chance for both addition and subtraction, but performed at chance levels for multiplication and division. The error patterns in that experiment suggested that for subtraction and addition, the right hemisphere does have some capacity for approximating the solution even when it is unable to generate the exact solution. Furthermore, right hemisphere accuracy in addition and subtraction was higher for problems with small operands than with large operands. An additional experiment assessed approximate and exact addition in the two hemispheres for problems with small and large operands. The left hemisphere was equally accurate in both tasks but the right hemisphere was more accurate in approximate addition than in exact addition. In exact addition, right hemisphere accuracy was higher for problems with small operands than large, but the opposite pattern was found for approximate addition. PMID- 17420035 TI - Impact of cypermethrin on stream fish populations under field-use in biotech soybean production. AB - The impact of biotech-soybean technology on freshwater ecosystems is being evaluated in the Rolling Pampas region, Argentina. The effect of cypermethrin, the main soybean insecticide, on low-order temperate-stream fish populations was investigated for two consecutive crop cycles under field-use conditions in biotech-soybean production. Cypermethrin was unable to induce mortality or behavioral effects on any of the fish species resident in a first-order stream across a crop field (pulsed acute exposure scenario) sprayed according to conventional practices. No spatially or temporally dependent effects were observed on population parameters (size-class structure, abundance, survival, sex and immature/mature ratio, condition factor) of resident or caged Cnesterodon decemmaculatus after spraying or rainfall events, not even one year after, at the beginning of the next crop cycle. Although cypermethrin was "very highly toxic" to C. decemmaculatus in laboratory water (96h-LC(50)=0.43microg/l), its toxicity was reduced in filtered (78%) and unfiltered (92%) stream water. Changes in LC(50) values were mainly correlated with the OC content of each water fraction (r(2)=0.99; p<0.01; n=9), showing that both DOC and TOC contributed proportionally to toxicity reduction. Protective effects of stream water (12-fold reduction LC(50) values) explained the lack of effects on fish populations in the field, despite cypermethrin water concentrations after spraying reached values comparable with the 96h-LC(50). Therefore, cypermethrin under field-use conditions in transgenic-soybean production represents a low risk of acute exposure for fish populations inhabiting low-order temperate-streams rich in TOC. The relationship between LC(50) and TOC could be a convenient way to improve risk estimation based on laboratory toxicity testing. PMID- 17420036 TI - Persistent chlorinated pesticides in fish species from Qiantang River in East China. AB - Thirteen organochlorine pesticides (OCPs) in 18 fish species from Qiantang River were firstly determined by GC-ECD. To elucidate the sources and the environment fate of these pollutants, water and sediment samples were also analyzed for OCPs contents. Total concentrations of OCPs in fish muscles ranged from 7.43 to 143.79 ng g(-1) wet weight (ww) with highest concentration recorded in sole fish (Cynoglossus abbreviatus), a benthic carnivore. The results indicated that carnivore fish have higher OCPs concentration than other fish with different feeding modes. OCPs concentration in fish was in the range of 1.86-5.85, 2.65 133.51 and 1.94-12.48 ng g(-1) for HCHs (alpha-, beta-, gamma-, delta-HCH), DDTs (p,p'-DDD, p,p'-DDE, p,p'-DDT, o,p'-DDD) and other OCPs (aldrin, diedrin, endrin, heptachlor, heptachlor expoide), respectively. The highest OCPs concentration in fish organs of four big fish species was found in brain of silver carp (Hypophthalmichthys molitrix), 289.26 ng g(-1) ww followed by kidney, liver, heart and gill. Among the OCPs analyzed, DDE, gamma-HCH and heptachlor were the predominant contaminants in fish muscle, which indicated that there was recent input of lindane. Significant correlation was observed between concentrations of DDTs and lipid content as well as between OCPs and lipid contents in fish species. Both field water bioconcentration factors (BCF) and sediment BCF showed a positive correlation with octanol-water partition coefficients (Kow) in the sole fish. PMID- 17420037 TI - Characterization of humic substances of different origin by means of mass spectrometry and neural networks. AB - Mass spectrometry fingerprinting of humic acids extracted from different soils has been carried out using laser desorption/ionization mass spectrometry (LDI-TOF MS). LDI-TOF MS provides characteristic mass spectra fingerprints for the humic acids of different origin. The information given in the fingerprints was evaluated for natural grouping trends in the samples by neural networks computing tools, such as self-organizing feature map (SOFM). This approach is efficient for recognizing patterns in the humic acids samples independently of their characteristic variability; variability characterizing natural products such as humic substances. The use of multi-layer perceptron artificial neural networks gave a successful classification of the samples. PMID- 17420038 TI - Trace metals assessment in water, sediment, mussel and seagrass species- validation of the use of Posidonia oceanica as a metal biomonitor. AB - The accumulation of trace metals (Cd, Co, Cr, Hg, Ni and Pb) was measured in water, sediment, the mussel Mytilus galloprovincialis and the seagrass Posidonia oceanica. Samples were collected in three locations of the north-western Mediterranean (Canari, Livorno and Porto-Torres) which present different levels and sources of human impact. Analyses in the different compartments (water, sediment, M. galloprovincialis and P. oceanica) have allowed to identify Canari as the most Cd, Co, Cr and Ni contaminated site; Livorno as the most Hg contaminated and Porto-Torres as the most Pb contaminated. Furthermore, for the first time, metal concentrations found in P. oceanica have been compared with those found in the water column, in the sediment and in the recognized metal bio indicator species M. galloprovincialis and the results obtained have led to the same conclusions. Thus, this study allows to validate the use of P. oceanica as metal biomonitor of coastal waters. PMID- 17420039 TI - Human menopausal and pregnant mare serum gonadotrophins in murine superovulation regimens for transgenic applications. AB - Superovulation is a fundamental procedure for generating transgenic rodents. While various methods exist, zygote yield/quality remain suboptimal, making these techniques open to refinement. All require a follicle stimulating and a luteinising effect. The former can be induced by pregnant mare serum gonadotrophin (PMSG) or other compounds like human menopausal gonadotrophin (HMG). While HMG can double zygote yield compared to PMSG, no study has compared their effects on embryo quality. Embryo yield could also be increased with PMSG: timing administration at estrus may further improve follicular recruitment. This study compared: (i) the efficacy of HMG/PMSG for producing viable embryos for microinjection; and (ii) the effect of HMG/PMSG administration at estrus on embryo yield. Whitten effect-induced estrous C57/Bl6xCBA F(1) hybrid mice were superovulated as follows: PMSG (day 1; 5 IU intraperitoneally) or HMG (days 1 and 2; 1 IU intramuscularly); all received human chorionic gonadotrophin (hCG) on day 3 (5 IU, intraperitoneally). Zygotes were retrieved following mating, morphologically assessed and microinjected with innocuous ZhAT1R construct (expressing LacZ reporter and human angiotensin II type 1 receptor) before transfer to pseudopregnant recipients. Pups were tested for the transgene by Southern blot. Neither HMG nor PMSG proved superior in improving embryo yield, morphology and short-term post-microinjection survival. However, HMG group micromanipulated embryos all failed to establish a pregnancy/generate transgenic pups, unlike their PMSG counterparts. While HMG can be used for superovulation, it appears to increase embryo vulnerability to the long-term effects of microinjection. Furthermore, the embryo yields associated with HMG can be replicated by timing PMSG injection to coincide with Whitten effect-induced estrus. PMID- 17420040 TI - Effects of follicular atresia and size on the developmental competence of bovine oocytes: a study using the well-in-drop culture system. AB - The effect of granulosa cell (GC) apoptosis and follicle size on the competence of bovine oocytes were studied using a well-in-drop (WID) oocyte/embryo culture system, which allows identification of follicular origin. Hatching rates of blastocysts did not differ (P>0.05) between oocytes cultured in the WID system (13%) and those cultured in the conventional group system (16%). Hatching rates of blastocysts were higher (P<0.05) in early atretic (17%) than in non-atretic (8%) and late atretic follicles (10%) of the same size (4-8mm), and in 6-8mm (22%) than in 4-5mm follicles (15%) at the early atretic stage. More oocytes (P<0.05) from late atretic (17%) than from non-atreteic (7%) or early atretic follicles (9%) of the same size (4-8mm) were arrested at Grade 1 cumulus expansion (only cells in the peripheral two layers began to expand). Similarly, more oocytes from 2 to 3mm follicles (30%) than from 6 to 8mm follicles (21%) at the same (late) atretic stage had Grade 1 cumulus expansion (P<0.05). Hatching blastocyst percentages of oocytes with Grade 3 (all layers of the cumulus except corona radiate cells expanded) or Grade 4 (full) cumulus expansion were higher in early atretic (20%) than in non-atretic (13%) or late atretic follicles (12%). Hatching blastocyst percentages of oocytes from follicles at the early atretic stage increased as cumulus expanded from Grade 2 (9%) to Grade 4 (27%). Regardless of the degree of follicle atresia, 72-76% of the floating cells in the follicular fluid (FF) were undergoing apoptosis. The floating cell density in FF was highly (r=0.6-0.7) correlated with oocyte developmental potency. In conclusion, the WID culture system was as efficient as group culture and allowed identification of follicular origin. Furthermore, the developmental potential of oocytes was affected by GC apoptosis, follicle size and cumulus expansion, and the floating cell density in FF could be used as a simple and non-invasive marker of oocyte quality. PMID- 17420041 TI - Detection of bovine viral diarrhea virus (BVDV) in single or small groups of preimplantation bovine embryos. AB - The objectives of this study were to develop techniques to detect BVDV associated with single or small groups of bovine embryos contained in small aliquots of medium using either virus isolation (VI) or real time quantitative polymerase chain reaction (RT-QPCR) assays. In vivo-derived and in vitro-produced bovine embryos at 7 d post-fertilization were exposed to SD-1, a high affinity strain of BVDV, for 2 h and then processed according to the International Embryo Transfer Society (IETS) guidelines prior to testing. Groups of five or two in vivo-derived embryos, and single in vivo-derived embryos, were VI positive for BVDV 100, 50, and 33% of the time, and were RT-QPCR positive 100, 75, and 42% of the time, respectively. The virus was detected by the VI technique in all of the groups of five or two in vitro-produced embryos and in all of the single in vitro-produced embryos, and it was detected in 100, 80, and 50%, using RT-QPCR. Techniques for RT-QPCR were sufficiently sensitive to detect 10 copies of viral RNA in a sample and to detect BVDV associated with single embryos. Application of this new technology, RT-QPCR, will facilitate additional studies to further assess the risk of transmission of BVDV through embryo transfer. PMID- 17420042 TI - Re: Stone et al.'s "effect of iodine on mercury concentrations in dental-unit wastewater". PMID- 17420043 TI - Influence of filler distribution on the color parameters of experimental resin composites. AB - OBJECTIVES: Simulating the optical properties of natural tooth would be the final goal for esthetic restorative materials. Filler distribution in resin composites determines the scattering in composite materials, which in turn would influence the color parameters such as lightness, chroma and hue. The objective of this study was to determine the influence of filler size and amount on the color parameters of experimental resin composites. METHOD: Color of 11 experimental resin composites with two different sized fillers (LG: 0.77 microm and SG: 0.50 microm) in 10-70 wt% was measured by a spectrophotometer. Color coordinates (CIE L*, a* and b*), chroma and hue angle were determined. Optical constants including scattering coefficient (S), absorption coefficient (K) and light reflectivity (RI) were calculated. To determine the influence of the amount of filler on the optical parameters, Pearson correlations between the amount of filler (%) and color parameters and optical constants were calculated. Correlations between the optical constants (S, K and RI) and color parameters were calculated (p<0.05). RESULTS: S value increased as the amount of filler increased. RI value generally increased as the amount of filler increased for LG filler group, and increased up to 40% filler for SG filler group. CIE L* value increased as the amount of filler increased in both of LG and SG filler groups. CIE L* value was highly correlated with S and RI values for both filler groups (r=0.961-0.974). CONCLUSION: Lightness was highly correlated with the amount of filler, S and RI values (r=0.932-0.974). But the correlation coefficients between the amount of filler and chroma/hue were moderate (r=0.406-0.827); therefore, pigmentation would be tried to simulate the color of resin composites to those of natural tooth. Optical properties of resin composites could be partly simulated to those of teeth by controlling the filler distribution. PMID- 17420044 TI - Investigations on C-H...pi interactions in RNA binding proteins. AB - We have investigated the roles played by C-Hcdots, three dots, centeredpi interactions in RNA binding proteins. There was an average of 55 C-Hcdots, three dots, centeredpi interactions per protein and also there was an average of one significant C-Hcdots, three dots, centeredpi interaction for every nine residues in the 59 RNA binding proteins studied. Main-chain to side-chain C-Hcdots, three dots, centeredpi interactions is the predominant type of interactions in RNA binding proteins. The donor atom contribution to C-Hcdots, three dots, centeredpi interactions was mainly from Phe, Tyr, Trp, Pro, Gly, Lys, His and Ala residues. The acceptor atom contribution to main-chain to side-chain C-Hcdots, three dots, centeredpi and side-chain to side-chain C-Hcdots, three dots, centeredpi interactions was mainly from Phe and Tyr residues. On the contrary, the acceptor atoms of Trp residues contributed to all the four types of C-Hcdots, three dots, centeredpi interactions. Also, Trp contributed both donor and acceptor atoms in main-chain to side-chain, main-chain to side-chain five-member aromatic ring and side-chain to side-chain C-Hcdots, three dots, centeredpi interactions. The secondary structure preference analysis of C-Hcdots, three dots, centeredpi interacting residues showed that, Arg, Gln, Glu, His, Ile, Leu, Lys, Met, Phe and Tyr preferred to be in helix, while Ala, Asp, Cys, Gly, Trp and Val preferred to be in strand conformation. Long-range C-Hcdots, three dots, centeredpi interactions are the predominant type of interactions in RNA binding proteins. More than 50% of C-Hcdots, three dots, centeredpi interacting residues had a higher conservation score. Significant percentage of C-Hcdots, three dots, centeredpi interacting residues had one or more stabilization centers. Seven percent of the theoretically predicted stabilizing residues were also involved in C-Hcdots, three dots, centeredpi interactions and hence these residues may also contribute additional stability to RNA binding proteins. PMID- 17420045 TI - A study of local effect and global effect on the microthermal bio-flows by molecular dynamics. AB - This study develops a hybrid numerical scheme based on a molecular dynamics (MD) algorithm and the GROMACS protein data bank to analyze the thermal bio-flow of alanine molecules in a microchannel. The numerical results show that the velocity profiles in the microchannel are highly dependent on both global effects, i.e. the effective channel width and local effects, i.e. the thermal boundary conditions. Specifically, the magnitude of the fluctuations observed in the velocity profiles increase as the channel width decreases or as the thermal boundary temperature increases. The results presented in this study provide useful information regarding suitable microchannel widths and operational temperatures for bio-chip devices and contribute a further understanding of basic human thermal bio-flow phenomena, particularly, regarding the correlation between the rate of local metabolism, burn and frostbite events, respectively. PMID- 17420046 TI - Social approach behaviors in oxytocin knockout mice: comparison of two independent lines tested in different laboratory environments. AB - Oxytocin mediates social affiliation behaviors and social memory in rodents. It has been suggested that disruptions in oxytocin contribute to the deficits in reciprocal social interactions that characterize autism. The present experiments employed a new social approach task for mice which is designed to detect low levels of sociability, representing the first diagnostic criterion for autism. Two lines of oxytocin knockout mice were tested, the National Institute of Mental Health line in Bethesda, and the Baylor/Emory line at the University of North Carolina in Chapel Hill. Similar methods were used for each line to evaluate tendencies to spend time with a stranger mouse versus with an inanimate novel object with no social valence. Adult C57BL/6J males were tested identically, as controls to confirm the robustness of the methods used in the social task. Comprehensive phenotyping of general health, neurological reflexes, olfactory and other sensory abilities, and motor functions was employed to assess both lines. No genotype differences were detected in any of the control measures for either line. Normal sociability, measured as time spent with a novel stranger mouse as compared to time spent with a novel object, was seen in both the NIMH and the Baylor/Emory lines of oxytocin null mutants, heterozygotes, and wild-type littermate controls. Normal preference for social novelty, measured as time spent with a second novel stranger as compared to time spent with a more familiar mouse, was seen in both the NIMH and the Baylor/Emory lines of oxytocin null mutants, heterozygotes, and wild-type littermate controls, with minor exceptions. Similar behavioral results from two independent targeted gene mutations, generated with different targeting vectors, bred on different genetic backgrounds, and tested in different laboratory environments, corroborates the negative findings on sociability in oxytocin mutant mice. Intact tendencies to spend time with another mouse versus with a novel object, in both lines of oxytocin knockouts, supports an interpretation that oxytocin plays a highly specific role in social memory, but is not essential for general spontaneous social approach in mice. PMID- 17420047 TI - Increased frequencies of glutathione-S-transferase (GSTM1 and GSTT1) null genotypes in Indian patients with chronic myeloid leukemia. AB - Inherited differences in the capacity of xenobiotic metabolizing enzymes might be an important factor in genetic susceptibility to cancer. Null genotypes of glutathione-S-transferases (GSTs) exhibit absence of enzymatic activity and are hypothesized to be at increased risk of developing cancers. The aim of the study was to examine whether null genotypes of GSTM1 and GSTT1 confer susceptibility to chronic myeloid leukemia (CML). We carried a case control study involving 80 consecutive North Indian CML patients (58 males, 22 females; age (mean+/-S.D.) 36.2+/-10.9 years) and 105 healthy individuals (59 males, 46 females; age (mean+/ S.D.) 36.8+/-11.3 years). Multiplex PCR was carried out to determine the frequency of GSTM1 and GSTT1 null genotypes. The relationship between GSTM1, GSTT1 genotypes and risk of CML was assessed by means of odds ratio (OR) with 95% confidence limits calculated by logistic regression. A test for trend (P(trend)) in increasing the risk of CML having more than one putative high-risk allele or genotype was evaluated by means of the chi-square test. There was no difference in the frequencies of the GSTM1 null genotype and the combined GSTM1 and GSTT1 null genotypes between patients and controls in the study. However, statistical significance was found with GSTT1 null genotype frequency in CML patients as compared to controls (16/80 (20%) versus 9/105 (8.5%); OR=2.67, 95% CI: 1.03 7.01). It projects a 2.67-fold increased risk for CML in individuals with GSTT1 null genotype as compared to those possessing both alleles of the gene. Our findings suggest that heritable GST status may influence the risk of developing CML. PMID- 17420048 TI - Effect of low-dose cytarabine and aclarubicin in combination with granulocyte colony-stimulating factor priming (CAG regimen) on the outcome of elderly patients with acute myeloid leukemia. AB - The aim of this study was to evaluate the efficacy and toxicity of low-dose cytarabine and aclarubicin in combination with granulocyte colony-stimulating factor (G-CSF) protocol in elderly patients with acute myeloid leukemia (AML). A total of 50 elderly patients including 8 aged over 70 years were enrolled. All patients were treated with CAG regimen including low-dose cytarabine (10mg/m(2) every 12h, days 1-14), aclarubicin (10mg every day, days 1-8), and G-CSF (200 microg/m(2) every day, days 1-14) priming. The overall response rate was 72.0%, and 29 of 50 (58.0%) patients achieved complete remission, including 23 of 35 (65.8%) with previously untreated AML, 6 of 15 (40.0%) with refractory, relapsed or secondary AML, 4 of 8 (50.0%) aged over 70 years, 4 of 10 (40.0%) with unfavorable cytogenetic aberrations. The early death rate was 7.6%. The median overall survival was 14 months. Myelosuppression was mild to moderate, severe nonhematologic toxicity was not observed. Thus CAG priming regimen as the induction therapy is well tolerated and effective in elderly patients with AML. PMID- 17420049 TI - A unique case of IgD-only splenic marginal-zone lymphoma with mutated immunoglobulin genes: ontogenetic implications. AB - Splenic marginal zone lymphoma (SMZL) is a rare indolent lymphoma subtype which accounts for less than 1% of all non-Hodgkin's lymphomas. We here report a unique case of IgD-only SMZL with mutated immunoglobulin variable region genes and discuss possible ontogenetic derivation. PMID- 17420050 TI - Alteration of adrenergic signals during peripheral blood stem cell mobilization induced by granulocyte colony-stimulating factor. PMID- 17420051 TI - Beta-interferon unbalances the peripheral T cell proinflammatory response in experimental autoimmune encephalomyelitis. AB - Interferon beta (IFNbeta) is a widespread therapy for multiple sclerosis (MS). We have analyzed some critical features of the T cell activation process in lymph nodes after IFNbeta treatment of experimental autoimmune encephalomyelitis (EAE) in SJL mice. Prevention of clinical signs and drastic reduction of perivascular infiltrates in the central nervous system (CNS) were accompanied by alterations in nuclear DNA binding activity levels of NFkappaB and Stat6 transcription factors in lymph node cells (LNC). A decrease of active NFkappaB subunits in treated animals correlated with lower levels of the cytoplasmic phosphorylated form of IkappaBalpha. Results also showed that nuclear DNA binding activity of Stat6 was increased by IFNbeta treatment, as were the cytoplasmic levels of phosphorilated Stat6 (P-Stat6). These high levels of P-Stat6 in IFNbeta-treated animals were accompanied by an increase of IL-4 expression levels measured by real time PCR. In vitro experiments with the IL-4 producing clone D10.G4.1 indicates that the IFNbeta-mediated IL-4 induction is not an effect exclusive to MBP-reactive cells, and suggest that it could be mediated by mRNA stability enlargement. On the other hand, IFNbeta treatment of EAE produced no significant changes in peripheral IFNgamma expression and a striking decrease of IL-17. These findings suggest that the inhibition of NFkappaB activity, the increase of IL-4 expression and its signaling transduction, and the decrease of IL-17 may cooperate to some of the antiinflammatory effects of IFNbeta on EAE. PMID- 17420052 TI - Expression sites of the collectin SP-D suggest its importance in first line host defence: power of combining in situ hybridisation, RT-PCR and immunohistochemistry. AB - Surfactant proteins A and D are pattern recognition molecules that play a role in pulmonary host defence. In this paper, we describe for the first time the expression and localisation of both collectins in various porcine tissues using a combination of in situ hybridisation (ISH), RT-PCR and immunohistochemistry (IHC). SP-D was expressed in several tissues including lung, tongue, intestinal tract, thymus, skin, gall bladder and lacrimal gland. Focal SP-D expression was detected in oesophagus, stomach, kidney, liver, prostate and spleen with both histological techniques. These tissues tested negative with RT-PCR. In contrast, SP-A expression was limited to the lung as measured by ISH and IHC. Interestingly, analysis by RT-PCR showed that thymus, trachea, jejunum and duodenum are positive for the presence of SP-A mRNA. We conclude that the combination of different methods can be advantageous if tissue-specific expression is studied. The importance of SP-D in innate immune defence of the pig is underlined by its expression at the potential ports of entry of pathogens. PMID- 17420053 TI - Confirmation of and explanations for elevated blood lead and other disorders in children exposed to water disinfection and fluoridation chemicals. AB - Silicofluorides (SiFs), fluosilicic acid (FSA) and sodium fluosilicate (NaFSA), are used to fluoridate over 90% of US fluoridated municipal water supplies. Living in communities with silicofluoride treated water (SiFW) is associated with two neurotoxic effects: (1) Prevalence of children with elevated blood lead (PbB>10microg/dL) is about double that in non-fluoridated communities (Risk Ratio 2, chi2p<0.01). SiFW is associated with serious corrosion of lead-bearing brass plumbing, producing elevated water lead (PbW) at the faucet. New data refute the long-prevailing belief that PbW contributes little to children's blood lead (PbB), it is likely to contribute 50% or more. (2) SiFW has been shown to interfere with cholinergic function. Unlike the fully ionized state of fluoride (F-) in water treated with sodium fluoride (NaFW), the SiF anion, [SiF6]2- in SiFW releases F- in a complicated dissociation process. Small amounts of incompletely dissociated [SiF6]2- or low molecular weight (LMW) silicic acid (SA) oligomers may remain in SiFW. A German PhD study found that SiFW is a more powerful inhibitor of acetylcholinesterase (AChE) than NaFW. It is proposed here that SiFW induces protein mis-folding via a mechanism that would affect polypeptides in general, and explain dental fluorosis, a tooth enamel defect that is not merely "cosmetic" but a "canary in the mine" foretelling other adverse, albeit subtle, health and behavioral effects. Efforts to refute evidence of such effects are analyzed and rebutted. In 1999 and 2000, senior EPA personnel admitted they knew of no health effects studies of SiFs. In 2002 SiFs were nominated for NTP animal testing. In 2006 an NRC Fluoride Study Committee recommended such studies. It is not known at this writing whether any had begun. PMID- 17420054 TI - Spatio-temporal constraints on upright children's coordination when hitting a moving target. AB - To determine if the spatial and/or temporal context affected the coordination patterns of children under five in upright interceptive tasks, 3-year-olds hit a ball dropped from a ramp from five heights (1.5, 2, 2.5, 3, and 3.5 m) to control drop time. We studied the initiation time of hitting a ball with a cinematic analysis of the different joint movements (foot, knee, hip, shoulder, and hand). Two patterns of joint coordination were observed according to the rolling height of the ball: a de-synchronized pattern for heights of 2 m or above where the foot was the first to move and the hand the last, and a "synchronized" pattern, for the lowest height (1.5 m) where all joints started moving at the same time due to a strong temporal pressure. These two coordination patterns highlight a functional adaptation of body joints according to the spatio-temporal constraints among children from the age of three. PMID- 17420055 TI - Infants' interactions with professional caregivers at 3 and 6 months of age: a longitudinal study. AB - This study longitudinally investigated the quality and stability of 64 infants' interactions with their professional caregivers in child care centers at 3 and 6 months of age, i.e., across the first 3 months after they entered child care. It was also examined whether the infants' negative emotionality (as rated by the mother) predicted the quality of the caregiver-infant interaction. The interactive behavior of the professional caregivers (sensitivity, cooperation) and the infants (responsiveness, involvement) was rated from videotapes recorded in three different caregiving situations, lasting about 25 min in total. In contrast to our expectation, the quality of the caregiver-infant interaction did not significantly increase across the first 3 months in child care. As expected, significant rank order stability was found for the quality of the caregivers' behavior over time. Also in accordance with our expectations, infants with higher negative emotionality scores experienced less sensitivity and cooperation in interactions with their primary professional caregivers at both ages. PMID- 17420056 TI - An improved plaque reduction virus neutralization assay for human metapneumovirus. AB - Virus-neutralizing antibodies against human metapneumovirus (hMPV) have been shown to be important indicators for protection in experimental animal models. An improved plaque reduction virus neutralization assay to detect hMPV-specific neutralizing antibodies was designed using two prototype recombinant hMPV strains expressing green fluorescent protein (GFP). These prototypes represented each of the main antigenic variants of hMPV, because antigenic variability could have implications for vaccine development. The utility of mutations in the F gene resulting in trypsin-independent replication was also tested. Although these mutant hMPV strains could replicate in the absence of trypsin, bigger plaque size was achieved with the addition of trypsin. Insertion of the GFP gene in the genome of hMPV did not affect replication of the virus in vitro. Plaques could be detected by measuring expression of GFP after 5 days by automated scanning. Ferret, hamster, and macaque sera positive for hMPV were compared in a conventional virus neutralization assay and the plaque reduction virus neutralization assay. The results obtained with the two assays were in agreement but the improved plaque reduction virus neutralization assay was faster, more suitable for high throughput testing, and 10-fold more sensitive than the conventional virus neutralization assay. PMID- 17420057 TI - Leading tonically active neurons of the striatum from reward detection to context recognition. AB - Tonically active neurons (TANs) in the primate striatum, which are presumed to be cholinergic interneurons, carry signals that are traditionally considered to be important for reward-related learning. Recent studies investigating the functional properties of TANs in behaving monkeys have shown that other factors beyond motivation can affect their responsiveness. There is now evidence that TAN responses reflect stimulus detection, movement control and recognition of a specific context, suggesting that these local circuit neurons contribute to different computations used in learning and action functions of the striatum. This is consistent with the view that TAN responses could represent an important component of the processes that are responsible for the ability to select the appropriate behavioral response to environmental events. PMID- 17420058 TI - The centrosome in neuronal development. AB - Establishment of polarity is an essential process during proliferation, migration and differentiation in neurons, and the signaling pathways leading to polarization of the cytoskeleton are topics of intense research. One key marker for cell polarity is the centrosome, also known as the microtubule-organizing center. Recent discoveries have shown that the position of the centrosome is precisely regulated during neurogenesis, migration and differentiation, leading to the segregation of cell fate factors, efficient nucleokinesis and directed neurite outgrowth, respectively. Here, we review recent advances in the understanding of this interesting organelle and propose a model whereby centrosome position, determined by extracellular factors, directs multiple aspects of neuronal development. PMID- 17420060 TI - A natural scavenger of peroxynitrites, rosmarinic acid, protects against impairment of memory induced by Abeta(25-35). AB - Peroxynitrite (ONOO(-))-mediated damage is regarded to be responsible for the cognitive dysfunction induced by amyloid beta protein (Abeta) in Alzheimer's disease (AD). In the present study, we examined the protective effects of rosmarinic acid (RA), a natural scavenger of ONOO(-), on the memory impairment in a mouse model induced by acute i.c.v. injection of Abeta(25-35). Mice daily received i.p. several doses of RA after the injection of Abeta(25-35). RA prevented the memory impairments induced by Abeta(25-35) in the Y maze test and novel object recognition task. RA, at the effective lowest dose (0.25mg/kg), prevented Abeta(25-35)-induced nitration of proteins, an indirect indicator of ONOO(-) damage, in the hippocampus. At this dose, RA also prevented nitration of proteins and impairment of recognition memory induced by ONOO(-)-i.c.v. injection. Co-injection of the non-memory-impairing dose of ONOO(-) with Abeta(25 35) blocked the protective effects of RA (0.25mg/kg). These results demonstrated that the memory protective effects of RA in the neurotoxicity of Abeta(25-35) is due to its scavenging of ONOO(-), and that daily consumption of RA may protect against memory impairments observed in AD. PMID- 17420059 TI - Olfactory bulbectomy increases food intake and hypothalamic neuropeptide Y in obesity-prone but not obesity-resistant rats. AB - Obese individuals often suffer from depression. The olfactory bulbectomy (OBX) model is an animal model of depression that produces behavioral, physiological, and neurochemical alterations resembling clinical depression. The OBX model was employed to assess depression-related changes in food intake in obesity-prone, Osborne-Mendel (OM) rats and obesity-resistant, S5B/Pl rats. OBX increased food intake in OM rats beginning 7 days following surgery, however, OBX did not alter food intake in S5B/Pl rats at any time point. Fourteen days following surgery, OBX significantly increased locomotor activity (total lines crossed and rears) in the openfield test in OM and S5B/Pl rats. Fifteen days following surgery, prepro neuropeptide Y (NPY) mRNA levels were significantly increased in the hypothalamus of bulbectomized OM rats and in the medial nucleus of the amygdala of bulbectomized OM and S5B/Pl rats. OBX decreased NPY Y2 receptor mRNA levels in the hypothalamus and medial nucleus of the amygdala in OM rats, while increasing NPY Y2 receptor mRNA levels in the medial nucleus of the amygdala of S5B/Pl rats. These data indicate that though both obesity-prone and obesity-resistant strains were susceptible to the locomotor effects of OBX, food intake and hypothalamic prepro-NPY mRNA were only increased in OM rats. Therefore, strain specific alterations in hypothalamic NPY may account for increased food intake in the obesity-prone rats following OBX, and suggests a potential mechanism to explain the comorbidity of obesity and depression. PMID- 17420061 TI - Mitochondria as a target for decavanadate toxicity in Sparus aurata heart. AB - In a previous in vivo study we have reported that vanadium distribution, antioxidant enzymes activity and lipid peroxidation in Sparus aurata heart are strongly dependent on the oligomeric vanadate species being administered. Moreover, it was suggested that vanadium is accumulated in mitochondria, in particular when V10 was intravenously injected. In this work we have done a comparative study of the effects of V10 and monomeric vanadate (V1) on cardiac mitochondria from S. aurata. V10 inhibits mitochondrial oxygen consumption with an IC(50) of 400 nM, while the IC(50) for V1 is 23 microM. V10 also induced mitochondrial depolarization at very low concentrations, with an IC(50) of 196 nM, and 55 microM of V1 was required to induce the same effect. Additionally, up to 5 microM V10 did inhibit neither F(0)F(1)-ATPase activity nor NADH levels and it did not affect respiratory complexes I and II, but it induced changes in the redox steady-state of complex III. It is concluded that V10 inhibits mitochondrial oxygen consumption and induces membrane depolarization more strongly than V1, pointing out that mitochondria is a toxicological target for V10 and the importance to take into account the contribution of V10 to the vanadate toxic effects. PMID- 17420062 TI - Optimizing gene suppression in schistosomes using RNA interference. AB - Schistosomes are parasitic platyhelminths that constitute an important public health problem globally. Here we describe optimized protocols for effectively suppressing gene expression in the intra-mammalian life stages of Schistosoma mansoni using RNA interference (RNAi). RNAi is a mechanism by which gene-specific double stranded RNA (dsRNA) triggers degradation of homologous mRNA transcripts. The gene encoding the cysteine protease cathepsin B (SmCB1 or Sm31) was targeted by exposing the parasites to dsRNA encoding part of the cathepsin B coding region. Suppression was measured using quantitative real time PCR. Electroporation as a mode of dsRNA delivery was substantially more efficient (100 1000-fold) than simply soaking the parasites in an equivalent dose. Soaking the parasites with dsRNA in the presence of different proprietary liposome preparations did not enhance gene suppression. In fact, all three reagents tested were variably toxic to the cultured schistosomes. Both long dsRNAs as well as synthetic short inhibitory RNAs (siRNAs) were effective at eliciting gene suppression. Different siRNAs exhibited variable efficiencies of suppression, perhaps reflecting differences in siRNA accessibility to the cathepsin B mRNA. Parasites cultured in vitro for 7 days or more following their emergence from the intermediate snail host were more susceptible to RNAi than those treated with dsRNA on the day of emergence (during the process of cercarial transformation into schistosomula). In addition, adult male and female parasites (49 days old) were susceptible to RNAi. Using fluorescein-labeled dsRNA to monitor the process, it was seen that in schistosomula (cultured for 7 days), electroporated dsRNA entered primarily through the mouth into the caecum while in young parasites (freshly emerged from snails) dsRNA appeared to enter primarily into the pre- and post-acetabular glands. The cathepsin B gene was significantly suppressed for up to 40 days after treatment suggesting that, as in some other organisms, the RNAi process can be amplified in schistosomes. PMID- 17420063 TI - The inherent property of polyhydroxyalkanoate synthase to form spherical PHA granules at the cell poles: the core region is required for polar localization. AB - Only the PHA synthase is required for formation of spherical intracellular PHA granules emerging at cell poles. This study aims to assign the polar targeting signal in the PHA synthase and to provide insight into molecular mechanisms of granule formation. Random in-frame insertion mutagenesis indicated dispensable and essential regions suggesting that only the N terminus (<100 aa) is dispensable and forms a random coil structure. The inactive PHA synthase (C319A) is still localized to cell poles, indicating that the nascent PHA chain does not serve as an anchor or signal for subcellular localization and granule formation. Deletion of the N terminus did neither affect subcellular localization nor PHA granule formation. The deletion of the hydrophobic C terminus (68 aa) did not impact on subcellular localization of the PHA synthase, but abolished PHA synthase activity. The structural protein PhaP1 was found to be not required for subcellular localization and initiation of granule formation. PhaP1 only localizes to the cell poles, when PHA granules are formed. These data suggested that the PHA synthase itself localizes to the cell poles via its core region (93 521 aa), which is structurally constraint and comprises the polar positional information for self-assembly of PHA granules at the cell poles. PMID- 17420064 TI - An amino acid substitution in the influenza A virus hemagglutinin associated with escape from recognition by human virus-specific CD4+ T-cells. AB - Influenza virus-specific CD4+ T-helper cells were cloned that recognized a virus strain isolated in 1981, but that failed to recognize more recent strains. The HLA-DR*1601-restricted epitope recognized was located in the hemagglutinin (HA(99 113)) and the naturally occurring A-->V substitution at position 106 was responsible for abrogating the recognition by HA(99-113)-specific CD4+ T-cells. This amino acid substitution was found in influenza A/H3N2 viruses that circulated between 1999 and 2005 and did not affect recognition by virus-specific antibodies. It was speculated that influenza viruses could evade recognition by virus-specific CD4+ T-cells, at least temporarily. PMID- 17420065 TI - Gender differences in the relationship between percent body fat (%BF) and body mass index (BMI) in Japanese children. AB - The purpose of the present study was to examine whether gender differences exist in the relationship between percent body fat (%BF) and body mass index (BMI) in Japanese children using a population-based cohort. Subjects are comprised of 187 boys and 163 girls aged 9-10, and 137 boys and 155 girls aged 12-13. Percent BF was measured using a bipedal biometrical impedance analysis (BIA) device. The relationship between %BF and BMI was investigated as a function of age and gender with Pearson's correlation coefficient. Strong linear relationships existed between %BF and BMI, especially in girls (9-10-year-old boys: r=0.779, P<0.0001; 9-10-year-old girls: r=0.975, P<0.0001; 12-13-year-old boys: r=0.786, P<0.0001; 12-13-year-old girls: r=0.975, P<0.000l). These results indicate that %BF can be predicted from BMI in Japanese children aged 9-10 and 12-13 years. The correlations in boys were not as strong as those observed in girls, that is, less variability was explained for girls than for boys. Further study will be necessary to ascertain whether the strong correlation seen among girls will be observed in different age or ethnic groups, and to ascertain the mechanism that produces this gender difference. PMID- 17420066 TI - Polymorphisms of cytosolic serine hydroxymethyltransferase and risk of lung cancer: a case-control analysis. AB - Suboptimal DNA repair capacity is a risk factor for cancer that may be modulated by dietary nutrient intake, and serine hydroxymethyltransferase (SHMT) participates in folate metabolism and synthesis of purines and pyrimidines needed for DNA repair. Therefore, we tested our hypothesis that genetic variants of the cytosolic SHMT (SHMT1) gene are associated with lung cancer risk. In a hospital based case-control study of 1032 non-Hispanic white lung cancer patients and 1145 matched cancer-free controls, we genotyped five common SHMT1 polymorphisms either in the promoter, exons, or 3'-untranslated regions. Although the genotype and allele frequency distribution of each SNP did not differ between cases and controls statistically significantly in the single-locus analysis, the rs638416 polymorphism in the promoter alone and the combined putative risk variant genotypes containing rs643333C, rs638416G, rs1979277T, rs3738G, and rs1979276C were associated with altered risk. Those carrying the combined 3+ risk variant genotypes had an increased risk of lung cancer (adjusted OR=1.65, 95% CI=1.05 2.57, compared with those having 0-1 risk genotypes; and OR=1.21, 95% CI=1.01 1.45, compared with those having 0-2 risk genotypes). The risk was more pronounced among older individuals (>61 years) or those having a low total folate intake or a high methionine intake. No evidence of interactions between the putative SHMT risk variant genotypes and the selected variables was found. These results suggest that SHMT1 variants may play a role in the etiology of lung cancer, and our findings need to be verified in larger prospective studies. PMID- 17420067 TI - Prognostic value of KIT expression in small cell lung cancer. AB - BACKGROUND: Small cell lung cancer (SCLC) is a very aggressive disease, with poor survival rates despite standard treatment with combination chemotherapy with or without radiotherapy. Further insights into the molecular biology of this malignant tumour are needed to improve the therapeutic approaches and outcome. KIT protein is expressed in SCLC, and its kinase activity has been implicated in the pathophysiology of many tumours, including SCLC. The purpose of this study was to evaluate the prevalence of KIT expression in patients with SCLC and its prognostic value. METHODS: We performed an inmunohistochemical analysis of 204 SCLC samples to determine KIT protein expression. The relationship between KIT expression and clinicopathological parameters was evaluated. Univariate and multivariate analyses were performed to define its prognostic significance. RESULTS: KIT expression was observed in 149 of 204 tumour tissues (73%). KIT expression was associated with advanced disease and with decreased incidence of bone metastases. No significant differences were observed for time to disease progression (TTP) (9.1% versus 6.2% at 3 years, p=0.6) or overall survival (OS) (10.7% versus 6.9% at 3 years, p=0.37) among patients with KIT positive versus negative tumours, respectively. Multivariate analysis showed that sex, tumour stage, albumin levels and response to therapy were the only independent predictors for survival. CONCLUSION: KIT protein is expressed in a high percentage of SCLC tumours. In our study population, however, the expression of KIT had no significant impact on survival. PMID- 17420068 TI - Angiokeratoma corporis diffusum in human beta-mannosidosis: Report of a new case and a novel mutation. AB - BACKGROUND: Human beta-mannosidosis, a rare disorder of oligosaccharide catabolism, results from a deficiency of beta-mannosidase activity. So far, mutational analysis has been performed in only seven families and revealed 11 mutations in the MANBA gene which encodes the enzyme beta-mannosidase. OBJECTIVES: We report here a 36-year-old Arab female with beta-mannosidosis who presented with mental retardation and multiple angiokeratomas. We describe in this patient a novel null mutation and review the previously reported MANBA gene mutations and their clinical correlations. METHODS: Histopathology, ultrastructural analysis, and enzyme assays were performed. Sequencing of cDNA and genomic DNA analysis was conducted in a search for a mutation in the MANBA gene. RESULTS: Histopathology of a skin biopsy specimen from the patient showed the characteristic findings of angiokeratoma. Electron microscopy showed cytoplasmic vacuolation. Enzymatic activity of beta-mannosidase in the patient's serum, leukocytes, and fibroblasts was less than 1% of control values. Sequencing of the MANBA cDNA revealed a G-->A transition in exon 6 at nucleotide position c.693, resulting in the formation of a stop codon (W231X). LIMITATIONS: Only one family was studied. CONCLUSIONS: A new case of human beta-mannosidosis is presented and the first MANBA gene mutation from Arab ancestry is reported. Reviewing the reported MANBA gene mutations does not reveal a clear genotype phenotype correlation. The importance of angiokeratoma corporis diffusum as the clue to the diagnosis of beta-mannosidosis and other lysosomal storage diseases is emphasized. PMID- 17420069 TI - Central oxytocin enhances antinociception in the rat. AB - The study aimed to investigate the effect of oxytocin on antinociception in the rat. The pain threshold was elevated by oxytocin following intraventricular (icv) or intrathecal injection (ith), and reduced by anti-oxytocin serum (icv or ith). But the pain threshold was not altered by intravenous injection (iv) of oxytocin or anti-oxytocin serum. Pain stimulation induced oxytocin concentration decrease in the hypothalamic supraoptic nucleus, and increase in the locus coeruleus, raphe magnus nucleus, caudate nucleus and spinal cord, but no change in the hypothalamic paraventricular nucleus and plasma. The results indicated that central, not peripheral oxytocin could enhance antinociception. PMID- 17420070 TI - Stereologic estimates of total spinophilin-immunoreactive spine number in area 9 and the CA1 field: relationship with the progression of Alzheimer's disease. AB - The loss of presynaptic markers is thought to represent a strong pathologic correlate of cognitive decline in Alzheimer's disease (AD). Spinophilin is a postsynaptic marker mainly located to the heads of dendritic spines. We assessed total numbers of spinophilin-immunoreactive puncta in the CA1 and CA3 fields of hippocampus and area 9 in 18 elderly individuals with various degrees of cognitive decline. The decrease in spinophilin-immunoreactivity was significantly related to both Braak neurofibrillary tangle (NFT) staging and clinical severity but not A beta deposition staging. The total number of spinophilin-immunoreactive puncta in CA1 field and area 9 were significantly related to MMSE scores and predicted 23.5 and 61.9% of its variability. The relationship between total number of spinophilin-immunoreactive puncta in CA1 field and MMSE scores did not persist when adjusting for Braak NFT staging. In contrast, the total number of spinophilin-immunoreactive puncta in area 9 was still significantly related to the cognitive outcome explaining an extra 9.6% of MMSE and 25.6% of the Clinical Dementia Rating scores variability. Our data suggest that neocortical dendritic spine loss is an independent parameter to consider in AD clinicopathologic correlations. PMID- 17420071 TI - Effect of age and caloric restriction on circadian adrenal steroid rhythms in rhesus macaques. AB - Dietary caloric restriction (CR) slows aging, extends lifespan, and reduces the occurrence of age-related diseases in short-lived species. However, it is unclear whether CR can exert similar beneficial effects in long-lived species, like primates. Our objective was to determine if CR could attenuate purported age related changes in the 24-h release of adrenal steroids. To this end, we examined 24-h plasma profiles of cortisol, and dehydroepiandrosterone sulfate (DHEAS) in young and old, male and female rhesus macaques (Macaca mulatta) subjected to either ad libitum (AL)-feeding or CR (70% of AL) for 2-4 years. Hormone profiles from young monkeys showed pronounced 24-h rhythms. Cortisol concentrations were higher in old males but not females, whereas DHEAS rhythms were dampened with age in both sexes. The cortisol rhythms of old CR males resembled those of young control males. However, CR failed to prevent age-related declines in DHEAS and further dampened DHEAS rhythms in both sexes. Apart from the partial attenuation of the age-related cortisol elevation in the old males, 24-h adrenal steroid rhythms did not benefit from late-onset CR. PMID- 17420072 TI - Double stranded RNA activated EIF2 alpha kinase (EIF2AK2; PKR) is associated with Alzheimer's disease. AB - Sporadic Alzheimer's disease (AD) appears to be the consequence of the interaction between combinations of genes and environmental factors (for example virus infections). To test this hypothesis, we are examining human genes relevant to herpes simplex virus type 1 (HSV-1) infection via genetic association studies in AD case-control samples. Recently, we found that a variant in TAP2, a major target used by HSV-1 to evade immune surveillance, is associated with AD. The present work analyses another gene involved in the host cell response to HSV-1, EIF2AK2 (eukaryotic translation initiation factor 2-alpha kinase 2; coding for PKR); PKR mediates the virus-induced shut-off of translation, and levels of activated PKR are high in the brains of AD patients. An EIF2AK2 SNP (rs2254958) located in the 5'-UTR region within an exonic splicing enhancer was found to be associated with AD. More specifically: the C allele was more commonly found in the patients and, compared to non-CC genotypes, the CC homozygotes showed earlier (around 3.3 years) onset of AD, especially in the absence of the APOE4 allele. These results further support the hypothesis that variants of human genes participating in HSV-1 infection modulate the susceptibility and/or clinical manifestations of AD. PMID- 17420073 TI - Synthesis of bridged piperazines with sigma receptor affinity. AB - Bridged piperazines 4 were designed as conformationally restricted piperazine sigma receptor ligands. The chiral pool synthesis started from (S)-glutamate, which was transformed in five reaction steps into the piperazinediones 5 bearing a propionic acid ester side chain. A two-step Dieckmann analogous cyclization provided the bicyclic ketones 7 as key intermediates. The alcohols 8 were prepared by LiAlH4 reduction of the ketones 7. NaBH4 reduction, Williamson ether synthesis and LiAlH4 reduction led to the methyl and benzyl ethers 12 and 13. High sigma1 affinity is attained when one large substituent is introduced either at N-8 or O-2. The most potent sigma1 ligand in this series of compounds is the methyl ether 12b with the N-butyl substituent (K(i)=13.2 nM, selectivity sigma2:sigma1 = 16). Moreover, the N-methyl derivatives 13a (sigma2: K(i)=30.4 nM) and 12a (sigma2 preference) represent promising starting points for the development of potent and selective sigma2 ligands. PMID- 17420074 TI - A conventional, inactivated oil emulsion vaccine suppresses shedding and prevents viral meat colonisation in commercial (Pekin) ducks challenged with HPAI H5N1. AB - The ongoing H5N1 epidemic is currently affecting a number of avian species, including waterfowl. These birds appear to have an important role as reservoirs of infection and comprehensive data on the efficacy of vaccination is currently lacking. The present paper reports the effect of a two dose vaccination programme with a conventional inactivated product on infection, lateral spread, shedding and presence of virus in commodities such as meat and viscera of Pekin ducks. Vaccination of this species appears to be efficacious in suppressing viral shedding, and preventing viraemia and lateral spread of infection to unvaccinated and vaccinated Pekin ducks. PMID- 17420075 TI - Combined DNA vaccination against three animal viruses elicits decreased immunogenicity of a single plasmid in mice. AB - In order to investigate whether combined DNA vaccines are an ideal way to combine antigens in a single vaccine formulation, we immunized mice with three plasmids (pVSG, pVgD and pVE2), respectively, encoding the antigen of foot-and-mouth disease virus (FMDV), pseudorabies virus (PRV) and classic swine fever virus (CSFV), either alone or in a combined vaccine regimen. We also investigated the immune responses induced by a series of mixtures in which three plasmids were mixed in pairs. Then we further immunized mice with three different plasmids in separate sites and preformed an adoptive transfer experiment. While being given alone, each of the vaccine plasmids induced significant virus-specific antibody responses and splenocytes proliferative activity. But reduced immunogenicity of the pVSG plasmid was found in combined DNA vaccination, no matter whether it was injected in a single or a separate site. Removal single plasmid (pVgD or pVE2) from combined DNA vaccine led to significant increase in the immunogenicity of the pVSG plasmid (P<0.05). And the induction of immune suppression was not mediated by suppressor T cells, as demonstrated by an adoptive transfer experiment. Furthermore, by boosting with whole virus protein of FMDV, mice primed with either pVSG alone or combined DNA vaccine produced statistically significant increase in the FMDV-specific antibody titers (P<0.05). But after boosting, FMDV-specific splenocytes proliferative activity of mice primed with combined DNA vaccine was even lower than that of mice primed with pVSG alone (P<0.05). Taken together, this study reflected the immunogenicity of a single plasmid may be decreased in combined DNA immunization strategy, which still needs to be carefully evaluated before practical application. PMID- 17420076 TI - Distribution of nuclear bomb Pu in Nishiyama area, Nagasaki, estimated by accurate and precise determination of 240Pu/239Pu ratio in soils. AB - Plutonium isotopes in forest soils collected in Nishiyama area, Nagasaki, were successfully determined by high resolution inductively coupled plasma mass spectrometry after the treatment with a microwave decomposition system. The (240)Pu/(239)Pu atom ratios observed in the samples in the Nishiyama area were obviously lower than the range of the global fallout. The low ratios (minimum 0.032) observed in Nishiyama area indicated the influence of detonation of the Pu nuclear weapon in 1945. Since the area is contaminated also by global fallout, the (240)Pu/(239)Pu atom ratio can be more sensitive indicator of bomb-derived Pu than Pu activity concentration. PMID- 17420077 TI - Naso-tracheal intubation to facilitate surgical access in parotid surgery. PMID- 17420078 TI - Apoptosis induced by uterine 24p3 protein in endometrial carcinoma cell line. AB - The biological functions and reaction pathways of lipocalins in mammalian system were sought. Mouse uterine 24p3 protein is a secreted lipocalin from mouse uterus. To evaluate the effect of uterine 24p3 protein on the reproductive system, endometrial carcinoma cell line (RL95-2) was an experimental target for achieving the in vitro study. The cells were treated with 0.75 microM dexamethasone (DEX) or under serum-free medium to mimic the stress environment for various time periods, then employing Western blot to measure the 24p3 protein secretion. It showed the time-dependent induction effect on 24p3 protein and suggested the level of protein secretion correlating to environmental stress. Furthermore, the supplementation of 24p3 protein to the medium accompanied the reduction of cell viability. It showed that the 24p3 protein may be a death factor under conditional media via PI and annexinV-FITC assay. Based on the autocrine hypothesis, we investigated the effect of 24p3 protein on cultured RL95 2 cells upon the 24p3 protein interaction. We have demonstrated significant increase in intracellular reactive oxygen species upon 24p3 protein interaction. While the changes of mitochondrial membrane potential and cytochrome c release from mitochondria occurred, the activities of caspase-8, -9 and -3 were found to have increased. The condensation of DNA was occurred suggesting that 24p3 protein induced irreparable DNA damage, which in turn triggered the process of apoptosis. It shows evidence for the direct effect of this protein on endometrial cells. These findings suggest that 24p3 protein creates an intracellular oxidative environment that induces apoptosis in RL95-2 cells. PMID- 17420079 TI - DNA damage and p53 in RAW264.7 cells induced by the spores of co-cultivated Streptomyces californicus and Stachybotrys chartarum. AB - Our recent studies have revealed that the co-cultivation of environmental microbes, Streptomyces californicus and Stachybotrys chartarum, potentiates the immunotoxic properties of the spores. In the present study, the spore-induced genotoxic potential of these microbes was investigated. Dose related differences in genotoxic and cytotoxic effects and in p53 level in mouse RAW264.7 macrophages were studied after 24h exposure to the spores of separately cultivated Streptomyces californicus or Stachybotrys chartarum alone, a simple spore-mixture of these microbes as well as to the spores of co-cultivated microbes. The genotoxic effect of the exposures was determined by the Comet assay and p53 level was analyzed by immunoblotting. Cytotoxicity was assessed by using flow cytometric analysis and also by the MTT test. The results revealed that the spores of co-cultivated microbes evoked DNA damage, p53 accumulation and cytotoxicity at a lower dose than the other exposures, and at the highest dose there was a 2.5-fold increase in DNA damage compared to control. In addition, the spores of Streptomyces californicus alone induced a 1.5-fold increase in DNA damage compared to control, dose dependent p53 accumulation and also extensive cytotoxicity. In contrast, the mixture of separately cultivated spores or the spores of Stachybotrys chartarum alone did not induce DNA damage with any tested dose although they triggered significant cytotoxicity and a slightly increased p53 level. Our results suggest that the detected genotoxic responses are the result of DNA damage in RAW264.7 cells by some genotoxically active metabolite(s) and the production of this compound was stimulated in Streptomyces californicus when it was co-cultivated with Stachybotrys chartarum. PMID- 17420080 TI - Chromosomal composition of micronuclei in mouse bone marrow treated with rifampicin and nicotine, analyzed by multicolor fluorescence in situ hybridization with pancentromeric DNA probe. AB - The mechanism underlying the induction of micronuclei induced by rifampicin and nicotine in mouse bone marrow was investigated by fluorescence in situ hybridization assay using mouse minor satellite DNA probe. Colchicine and mitomycin, known to be predominantly clastogenic and aneugenic, respectively were used as positive controls and these compounds produced the expected responses. In animals treated with different doses of rifampicin (10-320 mg/kg), the frequencies of micronucleated polychromatic erythrocytes (MNPCE) increased significantly after treatment with 160 and 320 mg/kg. Furthermore, rifampicin caused a significant depression of erythroblast proliferation at the high dose. At the two highest doses of 160 and 320 mg/kg rifampicin, a total of 0.96% and 1.44% MNPCE, respectively were found. Of the rifampicin-induced signal-positive MNPCE, an average of 58.1% of them was centromere-negative, reflecting the clastogenic activity of rifampicin. Correspondingly, about 41.9% of induced MNPCE were centromere-positive, representing the aneugenic activity of rifampicin. Eight and 16 mg/kg of nicotine induced 0.84% and 1.2% MNPCE, respectively, and of these an average of 29.5% showed one or more fluorescent signals, reflecting the predominant clastogenic activity of nicotine. The results obtained demonstrate that rifampicin induced both chromosome breakage and numerical chromosomal abnormalities, whereas nicotine induced one type of MNPCE and it could be considered a clastogenic agent. PMID- 17420081 TI - Conditioned flavor aversion and brain Fos expression following exposure to arsenic. AB - Recent advances in the knowledge of the cellular effects of arsenic have provided insights into the molecular mechanisms of arsenic-associated carcinogenesis, immunotoxicity and cardiovascular disease. In the present experiments we tested the hypothesis that the arrival of arsenic to the gastrointestinal (GI) tract is detected by the gut-brain axis, which includes hindbrain and forebrain nuclei activated by GI stimulation. As a marker of neuronal activation we measured Fos expression using immunohistochemistry. Because Fos expression in these nuclei is closely linked to the development of conditioned flavor aversion (CFA) we also tested the effect of arsenic on CFA. Our experiments indicate that arsenic ingestion is readily detected by the brain, as shown by increased Fos expression after oral administration of arsenic. Furthermore, the vagus nerve, which supplies information from the GI tract to the brain, is not involved in this response because a complete subdiaphragmatic vagotomy did not reduce the effect of arsenic on brain Fos expression, but enhanced this response. In parallel, arsenic ingestion is associated with a robust, dose-dependent CFA, which started at doses as low as 0.1 mg/kg body weight. In summary, these data indicate that arsenic given by oral administration is detected by the brain in low concentrations, and activates specific nuclei, which might trigger behavioral responses, such as CFA. PMID- 17420082 TI - Attitudes toward the performance of bystander cardiopulmonary resuscitation in Japan. AB - BACKGROUND: Early initiation of bystander cardiopulmonary resuscitation (CPR) improves the chances of successful resuscitation and survival. The importance of bystander CPR is attracting more interest, and there has been an increase in attendance at CPR training courses in Japan. However, there have been few reports regarding Japanese attitudes toward the performance of bystander CPR. The present study was performed to identify current Japanese attitudes toward bystander CPR compared to our previous study performed in 1998. METHODS AND RESULTS: Between February and March 2006, participants were asked about their willingness to perform CPR in five varying scenarios, i.e., performing CPR on a stranger, a trauma patient, a child, an elderly person, and a relative, and CPR techniques consisting of chest compression plus mouth-to-mouth ventilation (CC plus MMV) versus chest compression only (CC only). A total of 4223 individuals (male 50%) completed the questionnaire, including high school students, teachers, emergency medical technicians (EMTs), medical nurses, and medical students. About 70% of the subjects had experienced CPR training more than once. Only 10-30% of high school students, teachers, and health care providers reported willingness to perform CC plus MMV, especially on a stranger or trauma victim. In contrast, 70 100% of these subjects reported willingness to perform CC only, which was the same as in our previous study. The reasons for the unwillingness among laypeople to perform CC plus MMV were inadequate knowledge and/or doubt regarding whether they could perform the techniques effectively, while health care providers reported a fear contracting of a disease. CONCLUSIONS: Most laypeople and health care providers are unlikely to perform CC plus MMV, especially on a stranger or trauma victim, but are more likely to perform CC only, as also found in our previous study in 1998. These findings suggest that MMV training should be de emphasised and the awareness of CC alone should be emphasised because, for whatever reason, people do not want to perform MMV. PMID- 17420083 TI - The effect of a critical care outreach service and an early warning scoring system on respiratory rate recording on the general wards. AB - AIM: To determine whether the implementation of a Reading-Modified Early Warning Scoring (R-MEWS) system, is associated with an increased recording of respiratory rate (RR) in hospital inpatients, and whether the presence of a critical care outreach (CCO) service has a further impact on the recording of patient's vital signs. METHOD: Five annual point prevalence surveys of all adult, non-obstetric acute inpatients (n=2638) in two Hospitals (A and B) were carried out between 2001 and 2005. The R-MEWS system was implemented incrementally in both hospitals to include all study group patients, but a CCO service was only available in Hospital A. Data were collected on numbers of patients, routinely documented physiological observations and R-MEW score. RESULTS: Respiratory rate (RR) recording increased from 6.0% in the first survey to 77.9% in the last, which correlated with the incremental implementation of the R-MEWS system. Hospital A that had the CCO service showed a greater increase in RR recording than Hospital B with no CCO service. CONCLUSION: The introduction of an early warning scoring (EWS) was associated with improved respiratory rate recording, which may have been further enhanced by the presence of a CCO service. PMID- 17420084 TI - Outcome from paediatric cardiac arrest associated with trauma. AB - OBJECTIVE: To examine survival rates for paediatric trauma patients requiring cardiopulmonary resuscitation (CPR) in the pre-hospital setting, and to identify characteristics that may be associated with survival. DESIGN: Ten-year retrospective trauma database review. SETTING: An urban physician-led pre hospital trauma service serving a population of approximately 7.5 million, in the United Kingdom. PATIENTS: Eighty paediatric trauma patients (15 years or less) who received pre-hospital resuscitation following cardiorespiratory arrest between July 1994 and June 2004. INTERVENTION: Pre-hospital cardiopulmonary resuscitation. MAIN OUTCOME MEASURE: Survival to hospital discharge. RESULTS: Eighty children met inclusion criteria for the study. Nineteen (23.8%) were discharged alive from the emergency department and seven children (8.75%) survived to hospital discharge. Of the seven survivors, one had spinal cord injury. Two suffered asphyxial injury associated with blunt trauma and three sustained hypoxic insults following drowning or burns/smoke inhalation. In one patient with known congenital cardiac disease the cause of cardiac arrest was likely to have been medical. CONCLUSION: This study confirms the poor outcome for children requiring pre-hospital CPR following trauma. However, the results are better in this physician-attended group than in other studies where physicians were not present. They also suggest that cardiac arrest associated with trauma in children has a better outcome than in adults. In common with adults treated in this system, those patients with hypovolaemic cardiac arrest did not survive (Ann Emerg Med 2006;48:240-4). A large proportion of the survivors suffered hypoxic or asphyxial injuries. Targeted aggressive out-of-hospital resuscitation in certain patient groups can produce good outcomes. PMID- 17420085 TI - Platelet count and preoperative haemoglobin do not significantly increase the performance of established predictors of renal cell carcinoma-specific mortality. AB - OBJECTIVE: Anaemia and/or thrombocytosis were identified as independent predictors of poor survival in renal cell carcinoma (RCC). We tested the extent to which these markers worsen the prognosis in these patients. METHODS: Analyses targeted 1828 patients with renal cell carcinoma. Univariable, multivariable, and predictive accuracy analyses addressed RCC-specific mortality (RCC-SM). RESULTS: In univariable and multivariable analyses, both platelet count and preoperative haemoglobin level were statistically significant predictors of RCC-SM. However, neither platelet count nor preoperative haemoglobin level increased the combined multivariable accuracy of established RCC-SM (predictive accuracy gain=0.3%) predictors. CONCLUSIONS: Patients who present with severe anaemia or elevated platelets are at no higher risk of RCC-SM than that related to their stage, grade, histologic subtype, and Eastern Cooperative Oncology Group-Performance Status. PMID- 17420086 TI - Editorial comment on: Peripheral mechanisms of erectile dysfunction in a rat model of chronic cocaine use. PMID- 17420087 TI - Peripheral mechanisms of erectile dysfunction in a rat model of chronic cocaine use. AB - OBJECTIVE: To evaluate the peripheral mechanisms of erectile dysfunction (ED) in a rat model of triple-binge cocaine administration. METHODS: Adult male Sprague Dawley rats (n=24) were divided into two groups: group 1, control rats receiving vehicle (saline); group 2, rats receiving binge cocaine injections. After completion of triple-binge cocaine or saline injections, both groups underwent an in vivo, neurogenic-mediated erectile response protocol to assess intracavernosal pressure (ICP). Penile endothelin-A and -B receptors (ET(A)R and ET(B)R), plasma levels of big endothelin-1 (big-ET-1), and endothelial nitric oxide synthase (eNOS) protein expression were assessed. To analyze nitric oxide (NO) production, we measured plasma nitrate-nitrite levels and quantitated myeloperoxidase (MPO) activity in cavernosal tissues to determine reactive oxygen species generation. Endothelium-dependent and -independent relaxation responses were evaluated in vitro. Data were analyzed with Student t test. RESULTS: Triple-binge cocaine administration caused significantly decreased erectile responses as measured by ICP in vivo. Plasma big-ET-1 levels were significantly increased in the triple binge cocaine treatment group compared with control animals. In the penis, triple binge cocaine administration significantly increased ET(A)R expression compared with saline controls, while ET(B)R expression was not altered. Cocaine-treated rats had significantly decreased eNOS expression and NO production. The activity of tissue MPO was significantly increased in the cocaine group compared with control rats. Organ bath studies demonstrated that triple-binge cocaine resulted in a 64% reduction in maximal relaxation compared with the control group. CONCLUSION: This study demonstrates that triple-binge cocaine administration significantly reduces erectile function in rats. The pathophysiologic mechanisms that are likely involved include increased plasma big-ET-1 levels, increased penile ET(A)R expression, increased penile MPO activity, and reduced penile eNOS expression. PMID- 17420088 TI - Adjuvant therapy with sorafenib in bone metastases bilateral renal carcinoma: a case report. AB - A 69-yr-old man with bilateral and metastatic renal cell carcinoma developed progressive disease after interleukin-2 and interferon therapy. He was submitted to radical left nephrectomy, right nephron-sparing surgery, and bone metastasis removal, followed by therapy with sorafenib. At 12-mo follow-up there was a significant improvement in patient performance status and no evidence of clinical progression. PMID- 17420089 TI - Editorial comment on: Platelet count and preoperative haemoglobin do not significantly increase the performance of established predictors of renal cell carcinoma-specific mortality. PMID- 17420090 TI - Comments on "Removal of lead from aqueous solution using Syzygium cumini L.: equilibrium and kinetic studies". AB - This letter reports the importance and advantages of the constraints in the Redlich-Peterson isotherm exponent. PMID- 17420091 TI - A model for treating polluted air streams in a continuous two liquid phase stirred tank bioreactor. AB - Biological air treatment systems have been widely under investigation in recent years. Inclusion of non-biodegradable organic solvents to these systems is a way to improve the biotic removal capacity of the systems. In this article the process of absorption and biodegradation of a hydrophobic organic compound in a two liquid phase stirred tank bioreactor has been modeled. Using the model it has been shown that the inclusion of an organic solvent is advantageous if certain conditions are met. Some simulation examples showed that the usefulness of adding an organic solvent to the system depends on kinetic parameters of biological reactions and mass transfer coefficients of pollutants and oxygen between the air and liquid phases. Since different factors influence the process, the usefulness of including an organic solvent to the system should be checked in each special case. The simple model presented in this article can help the prediction of the effect of amending a solvent to the bioreactor under a set of given conditions. PMID- 17420092 TI - Influence of anionic species on uranium separation from acid mine water using strong base resins. AB - The presence of uranium and other elements in high concentrations in acid mine drainage at Pocos de Caldas Uranium Mine (Brazil) is a matter of concern. The acid water pH is around 2.7, the uranium concentration is in the range of 6-14 mg L(-1), sulfate concentration near 1400 mg L(-1), fluoride 140 mg L(-1) and iron 180 mg L(-1). In this solution, where sulfate is present in elevated concentrations, uranium is basically in the form of UO(2)(SO(4))(3)(4-). This study investigated the separation of uranium from the other anions present in the acid water under batch and column mode using ion exchange technique. The pH studied was 2.7 and 3.9. Two strong base anionic resins were tested. The influence of ions, commonly found in acid waters like sulfate and fluoride, on ion exchange process was also assessed. Equilibrium studies were carried out to determine the maximum adsorption capacities of the resins. The resins showed a significant capacity for uranium uptake which varied from 66 to 108 mg g(-1) for IRA 910U and 53 to 79 mg g(-1) for Dowex A. The results also showed that SO(4)(2 ) is the most interfering ion and it had a deleterious effect on the recovery in the pH range studied. Fluoride did not affect uranium removal. PMID- 17420093 TI - Removal of tungsten oxyanions from industrial wastewater by precipitation, coagulation and flocculation processes. AB - Tungsten removal from industrial wastewater by precipitation, coagulation and flocculation processes using ferric chloride is reported. Suitable process conditions (pH, ferric chloride concentration) were established in jar tests performed with wastewater samples. Alkaline wastewater was treated with ferric chloride and pH was adjusted to various points using sulphuric acid. Tungsten removal was found to be most efficient (98-99%) in acidic conditions (pH<6). The process conditions were also found to be suitable for operation of an industrial scale wastewater treatment facility. More than 97% of tungsten were removed and the residual concentration was smaller than 10 ppm. PMID- 17420094 TI - The effects of selected parameters on the nitric oxide removal by biofilter. AB - A bench-scale biofilter was used to demonstrate the treatability of off-gas containing nitric oxide (NO) by examining selected operational parameters. After 6 days of operation, the biofilter reached to a steady state and NO reduction was significant, reducing from 200 ppm to 95 and 40 ppm after 6 and 40 days of continuous operation. The oxygen concentrations in the inlet would affect NO removal performance significantly; as oxygen content decreasing from 6% to 0%, the NO removal efficiency increased from 55% to 99%, indicating that oxygen inhibited the progress of denitrification. NO removal was inversely proportional to inlet NO concentration, removal efficiency decreased from 88% to 40 % as NO concentration increasing from 60 to 500 ppm. Column height would significant effect on the NO removal efficiency, under column height=6.5m and O(2)=6% conditions, 90% of removal efficiency was achievable. The effect of glucose added into biofilter would significantly enhance the NO removal efficiencies for both anaerobic and aerobic conditions of which 99% and 55%, respectively. PMID- 17420095 TI - Comments on "equilibrium and kinetic studies for the biosorption system of copper(II) ion from aqueous solution using Tectona grandis L.f. leaves powder". AB - This letter reports the way to solve the Redlich-Peterson isotherm. In addition the importance of constraints for the Redlich-Peterson isotherm exponent was discussed in detail. PMID- 17420096 TI - Acute nicotine enhances c-fos mRNA expression differentially in reward-related substrates of adolescent and adult rat brain. AB - A number of studies have demonstrated that adolescent rodents are more sensitive to the rewarding effects of nicotine compared to adults. To help determine the potential brain circuitry involved, we investigated the effect of acute nicotine administration (0.4 or 0.8mg/kg, s.c.) on the expression of c-fos mRNA in the brains of adolescent (P35) and adult (P67-70) male Wistar rats using in situ hybridization. Nicotine administration increased c-fos mRNA expression in several brain regions, including the central amygdala, locus coeruleus, nucleus accumbens core, paraventricular nucleus of the hypothalamus and lateral septum of adolescent and adult rats. Nicotine increased c-fos mRNA expression more robustly in the bed nucleus of the stria terminalis, nucleus accumbens shell and ventral tegmental area in adolescent rats. The current results suggest that nicotine may have greater activational effects in brain regions associated with reward in adolescent rats and may help to explain the differences between adolescents and adults in behavioral responses to nicotine. PMID- 17420097 TI - Effects of engrafted neural stem cells derived from GFP transgenic mice in Parkinson's diseases rats. AB - This study evaluated the therapeutic effect of neural stem cells (NSCs) transplanted into Parkinson's disease (PD) rats. NSCs were identified in vitro, then engrafted into the striatum of the PD rats. The rotational behavior was evaluated 1, 2, 4 and 6 weeks. A significant rotational behavior improvement was observed in PD rats subjected to cell transplantation. Transplanted NSCs not only express Nerve growth factor and Neurotrophin-3 in vitro, but also survive and partly differentiate into tyrosine hydroxylase (TH) positive cells in vivo. The results show that NSCs could be effective for PD treatment and the mechanisms might involve the neurotrophin expression and the neural differentiation. PMID- 17420098 TI - Lesions of the medial prefrontal cortex prevent the induction but not expression of morphine-induced behavioral sensitization in mice. AB - Repetitive exposure to morphine induces behavioral sensitization, which is supposed to involve in the process of addiction to drugs. As the underlying neuropharmacological mechanisms and anatomical substrates are considerably different between different drugs and different phases of behavioral sensitization, this study was designed to investigate the roles of the medial prefrontal cortex in the induction and expression of morphine-induced behavioral sensitization in mice. In experiments 1 and 2, mice were dosed with morphine (10 mg/kg, i.p.) once daily for 7 continuous days after induction of sham or kainic acid lesions of the medial prefrontal cortex. Locomotor activity was measured on days 1 and 7 to test acute morphine-induced hyperactivity and the induction of behavioral sensitization. In experiment 3, mice were subjected to surgery after behavioral sensitization was achieved. The mice were challenged with morphine 7 days later to evaluate the expression of behavioral sensitization. The results showed that lesions of the medial prefrontal cortex (mPFC) blocked acute morphine induced hyperactivity and the induction of behavioral sensitization, but the lesions had no effect on the expression of behavioral sensitization. These results provide the first direct evidence that the mPFC may be involved in the induction, but not the expression, of morphine-induced behavioral sensitization. PMID- 17420099 TI - Association of IL10 promoter polymorphism in Italian Alzheimer's disease. AB - Recent studies have reported a genetic association between single nucleotide polymorphisms (SNPs) in the promoter region of Interleukin (IL) 10 and Alzheimer's disease (AD) with conflicting results. To further investigate the proposed association and to clarify the role of cytokines as a potential cause for AD susceptibility, we analyzed genotypes, allele distributions and haplotypes of IL10 promoter polymorphisms -1082 (rs1800896) and -819 (rs1800871) in an Italian sample of 222 sporadic AD patients and 179 normal controls. All 401 subjects were also genotyped for the Apolipoprotein E (ApoE) polymorphism. We reported a positive association between the -819T/C polymorphism and AD. Moreover, we found a significant difference for this SNP in the ApoE varepsilon4 non-carrier AD patients compared to the ApoE varepsilon4 non-carrier control group. For the -1082A genotype and allele distribution, no significant association was found in AD patients, although it was detected within the AT haplotype. Our results indicate that IL10 polymorphisms may be involved in the risk of developing AD. PMID- 17420100 TI - Ethanol-induced oxidative stress is mediated by p38 MAPK pathway in mouse hippocampal cells. AB - It has been known that ethanol causes neuronal cell death through oxidative stress. Ethanol itself and reactive oxygen species (ROS) produced by ethanol modulate intracellular signaling pathways including mitogen-activated protein kinase (MAPK) cascades. This study was conducted to examine the impact of ethanol on MAPK signaling in HT22 cells. Ethanol (100 and 400mM) caused activation of ERK, p38 MAPK, and JNK. ERK activation occurred in early time and p38 MAPK activation was evident when ERK activation was diminished. Specific inhibitor of p38 MAPK (SB203580) protected HT22 cells against ethanol, which was accompanied by an inhibition of ROS accumulation. However, inhibitors of ERK (U0126) and JNK (SP600125) had no effects on ethanol-induced neuronal cell death when they are treated with ethanol for 24h. These results suggest that p38 MAPK may have important roles in ROS accumulation during ethanol-induced oxidative stress in HT22 cells. PMID- 17420101 TI - Clinical, haematological and therapeutic studies on tropical theileriosis in water buffaloes (Bubalus bubalis) in Egypt. AB - Thirty buffaloes naturally infected with Theileria annulata and 10 parasitologically free controls were used to determine the potential clinical, haematological and therapeutic impact of tropical theileriosis in Egypt. The clinical signs in the infected buffaloes were pyrexia (40.5-41.5 degrees C), enlargement of superficial lymph nodes, slight nasal and ocular discharges, salivation, anaemia and respiratory distress. Eye lesions also were recorded. There was a significant decrease in erythrocyte counts and haemoglobin content and a significant decrease in total leucocyte counts in infected buffaloes compared to controls. Early treatment with buparvaquone was 100% effective in eliminating the protozoan parasites from the blood and lymph nodes and led to an improvement in the clinical state whereas treatment in the later stages of the disease whilst eliminating the parasites failed to improve the clinical condition of the animal. PMID- 17420102 TI - Occurrence of phantom genitalia after gender reassignment surgery. AB - Transsexuals are individuals who identify as a member of the gender opposite to that which they are born. Many transsexuals report that they have always had a feeling of a mismatch between their inner gender-based "body image" and that of their body's actual physical form. Often transsexuals undergo gender reassignment surgery to convert their bodies to the sex they feel they should have been born. The vivid sensation of still having a limb although it has been amputated, a phantom limb, was first described by Weir Mitchell over a century ago. The same phenomenon is also occurs after amputation of the penis or a breast. Around 60% of men who have had to have their penis amputated for cancer will experience a phantom penis. It has recently been shown that a significant factor in these phantom sensations is "cross-activation" between the de-afferented cortex and surrounding areas. Despite this it also known that much of our body image is innately "hard-wired" into our brains; congenitally limbless patients can still experience phantom sensations. We hypothesise that, perhaps due to a dissociation during embryological development, the brains of transsexuals are "hard-wired" in manner, which is opposite to that of their biological sex. We go on to predict that male-to-female transsexuals will be much less likely to experience a phantom penis than a "normal" man who has had his penis amputated for another reason. The same will be true of female-to-male transsexuals who have had breast removal surgery. We also predict that some female-to-male transsexuals will have a phantom penis even although there is not one physically there. We believe that this is an easily testable hypothesis, which, if correct, would offer insights into both the basis of transsexuality and provide farther evidence that we have a gender specific body image, with a strong innate component that is "hard-wired" into our brains. This would furnish us with a better understanding the mechanism by which nature and nurture interact to link the brain-based internal body image with external sexual morphology. We would emphasise here that transsexuality should not be regarded as "abnormal" but instead as part of the spectrum of human behaviour. PMID- 17420103 TI - Equine learning behaviour: what we know, what we don't and future research priorities. PMID- 17420104 TI - Stereospecific hydrolysis of a phosphoramidate as a model to understand the role of biotransformation in the neurotoxicity of chiral organophosphorus compounds. AB - Calcium-dependent and EDTA-resistant hydrolyses of R and S isomers of O-hexyl O 2,5-dicholorophenyl phosphoramidate (HDCP) were observed in serum and subcellular fractions of liver, kidney and brain from hen, rat and rabbit. In serum, the Ca(2+)-dependent hydrolysis was much higher in rabbit than in other species. Liver showed a higher activity than kidney and brain. The S-HDCP isomer was hydrolysed to a higher extent than the other isomer. The fact that this stereospecificity favours the S-isomer is more clearly observed in rabbit serum, and in rat and rabbit liver particulate fractions. In such tissues and species, the EDTA-resistant hydrolysis was not stereospecific. Soluble fractions of rat brain and of hen liver, kidney and brain, showed a lower total activity but with a higher proportion of EDTA-resistant activity and a higher hydrolysis of the R HDCP isomer. The Ca(2+)-dependent stereoselective biodegradation of S-HDCP is dominant in the most active tissues in rabbit and rat. It can therefore be concluded that S-HDCP would be biodegraded faster than R-HDCP. Furthermore, R HDCP is the isomer that will remain at a higher proportion to be available for interaction with the target of neurotoxicity. PMID- 17420105 TI - Two days of pulsatile GnRH infusion beginning 4 days before weaning in sows initiates a wave of follicular growth that is not sustained after weaning. AB - Intervals to estrus and ovulation in weaned sows depend partially on the diameter of ovarian follicles at weaning. The objective was to determine if follicular diameter in sows could be increased by a 48h period of GnRH infusion before weaning and whether this pre-weaning growth would advance follicular development after weaning. The posterior vena cava was cannulated in eight sows at 10+/-1 day after farrowing. Sows were randomly assigned to receive intravenous treatment with either 2mL of GnRH (1microg/mL; n=4) or 2mL of saline (n=4) every 0.5h for 48h beginning 94h before weaning. The average follicular diameter and the number of follicles within diameter classes were determined daily by ultrasonography. Serum LH concentrations increased on the first infusion day but serum LH was equal to control on the last infusion day (P<0.077). The GnRH infusion increased the average diameter of ovarian follicles (P<0.001). Serum estradiol increased (P<0.001) and serum FSH decreased (P<0.016) coincident with GnRH-induced follicular development but these changes were reversed within 24h after the end of the infusion period. Follicles that grew in response to GnRH regressed and were replaced by a new population of follicles within 4 days after weaning. Within the experimental model for the present study, a GnRH infusion increased follicular growth in lactating sows but follicles could not be sustained beyond the end of GnRH infusion. PMID- 17420106 TI - CT scans and 3D reconstructions of Florida manatee (Trichechus manatus latirostris) heads and ear bones. AB - The auditory anatomy of the Florida manatee (Trichechus manatus latirostris) was investigated using computerized tomography (CT), three-dimensional reconstructions, and traditional dissection of heads removed during necropsy. The densities (kg/m3) of the soft tissues of the head were measured directly using the displacement method and those of the soft tissues and bone were calculated from CT measurements (Hounsfield units). The manatee's fatty tissue was significantly less dense than the other soft tissues within the head (p<0.05). The squamosal bone was significantly less dense than the other bones of the head (p<0.05). Measurements of the ear bones (tympanic, periotic, malleus, incus, and stapes) collected during dissection revealed that the ossicular chain was overly massive for the mass of the tympanoperiotic complex. PMID- 17420107 TI - Development and application of electrospray-active derivatization reagents for hydroxysteroids. AB - New derivatization reagents, 1-(2,4-dinitro-5-fluorophenyl)-4-methylpiperazine (PPZ) and 4-(4-methyl-1-piperazyl)-3-nitrobenzoyl azide (APZ), were developed for the liquid chromatography-electrospray ionization-mass spectrometric (LC-ESI-MS) determination of steroids having a hydroxy group. PPZ reacted with a phenolic hydroxy group in estrogens. After quaternarization of the PPZ-estrogens with methyl iodide, the resulting derivatives provided more than a 2000-fold higher sensitivity compared to the intact estrogens. After derivatization of steroids having an alcoholic hydroxy group (5-ene-steroids or 5alpha-reduced steroids) with APZ followed by methylation, their detection responses increased more than 500 times. These derivatization procedures coupled with LC-ESI-MS/MS were successfully used for the determination of estrogens in the serum and prostatic 5alpha-dihydrotestosterone. PMID- 17420108 TI - Identification of an unknown trace-level impurity in bulk drug of Seroquel by high-performance liquid chromatography combined with mass spectrometry. AB - An impurity was detected in bulk drug Seroquel at about 0.4% level by the reversed-phase high-performance liquid chromatography with UV detection. The accurate mass of impurity was measured by FTICR equipped with electrospray ionization interface, and the structure of impurity was characterized on the basis of the on-line multi-stage mass spectrometric evidences. The proposed structure was further confirmed by multi-stage mass spectrometry of Seroquel and four related compounds. PMID- 17420109 TI - Outcomes of educational interventions in type 2 diabetes: WEKA data-mining analysis. AB - OBJECTIVE: To analyze which factors contribute to improvement in glycemic control in educational interventions in type 2 diabetes reported in randomized controlled trials (RCT) published in 2001-2005. METHODS: Papers were extracted from Medline and Scopus using educational intervention and adults with type 2 diabetes as keywords. Inclusion criteria were RCT design. Data were analyzed with a data mining program. RESULTS: Of 464 titles extracted, 21 articles reporting 18 studies met the inclusion criteria. Data mining showed that for initial glycosylated hemoglobin (HbA1c) level < or = 7.9% the diabetes education intervention achieved a small change in HbA1c level, or from +0.1 to -0.7%. For initial HbA1c > or = 8.0%, a significant drop in HbA1c level of 0.8-2.5% was found. Data mining indicated that duration, educational content and intensity of education did not predict changes in HbA1c levels. CONCLUSION: Initial HbA1c level is the single most important factor affecting improvements in glycemic control in response to patient education. Data mining is an appropriate and sufficiently sensitive method to analyze outcomes of educational interventions. Diversity in conceptualization of interventions and diversity of instruments used for outcome measurements could have hampered actual discovery of effective educational practices. PRACTICE IMPLICATIONS: Participation in educational interventions generally seems to benefit people with type 2 diabetes. Use of standardized instruments is encouraged as it gives better opportunities to identify conclusive results with consequent development of clinical guidelines. PMID- 17420110 TI - Separation of protein and lactose intake over meals dissociates postprandial glucose and insulin concentrations and reduces postprandial insulin responses in heavy veal calves. AB - The present study examined, at identical daily nutrient intakes, the impact of separating protein and lactose intakes across two daily meals on the metabolic and endocrine status in heavy veal calves. Calves were assigned to one of six degrees of separating protein and lactose over the two meals (termed nutrient synchrony, SYN 1-6; 6 calves/treatment). They were fed the protein-rich (P-)meal and the lactose-rich (L-)meal at 06:00 and 18:00h, respectively, or vice versa. At SYN 1, calves were fed with 50% of the daily protein and 50% of the daily lactose intake in each meal. Protein and lactose were iso-energetically exchanged between the two daily meals from SYN 1 to 6. At SYN 6, 85% of the daily protein and 20% of the daily lactose was fed in the P-meal and the remainder in the L meal. Blood samples were collected hourly during 24h. Mean 24h glucose concentrations increased and insulin concentrations decreased from SYN 1 to 6. Postprandial 5h areas under concentration curves (AUC(0-5h)) of glucose increased with increasing meal lactose content. AUC(0-5h) of non-esterified fatty acids increased after P- and L-meals from SYN 1 to 6. Urea concentrations increased after L-meals from SYN 1 to 6, but decreased after P-meals from SYN 1 to 6. Insulin AUC(0-5h) decreased after L-meals and after P-meals from SYN 1 to 6. Nutrient asynchrony did not affect insulin-like growth factor-1, glucagon, growth hormone, leptin, 3,5,3'-triiodothyronine and thyroxine. In conclusion, separation of protein and lactose intake over meals inhibited insulin responses to a lactose rich meal in heavy veal calves despite high plasma glucose concentrations. PMID- 17420111 TI - Significance of thyroid blood flow as a predictor of methimazole sensitivity in untreated hyperthyroid patients with Graves' disease. AB - OBJECTIVE: The peak systolic velocity (PSV) of the inferior thyroid artery (ITA) is increased in untreated hyperthyroid patients with Graves' disease (GD). We investigated the clinical significance of the ITA-PSV and its determinants in hyperthyroid GD patients. PATIENTS AND METHODS: ITA-PSV, together with thyroid volume, was measured by ultrasonography in untreated hyperthyroid GD patients (n=49) and healthy subjects (n=22). Established markers of GD activity such as TSH receptor antibody (TRAb), thyroid stimulating antibody (TSAb), vascular endothelial growth factor (VEGF) and immunoglobulin E (IgE) were simultaneously determined. RESULTS: ITA-PSV, thyroid volume, VEGF and IgE were significantly higher in hyperthyroid GD patients than in normal subjects. ITA-PSV in hyperthyroid GD patients was correlated positively with serum levels of FT(3), FT(4) and IgE, smoking index and thyroid volume, and negatively with total, HDL- and LDL-cholesterols, but did not correlate significantly with age, triglyceride, TRAb, TSAb or VEGF. In stepwise regression analysis, ITA-PSV showed significant positive and negative associations with IgE and LDL-cholesterol, respectively, in hyperthyroid GD patients. In the pre-treatment hyperthyroid state, FT(4) and ITA PSV, but not IgE, were found to be significantly and positively associated with the maintenance dose of methimazole (MMI) required to keep serum TSH within normal range for at least 12 months. CONCLUSION: These results suggest that ITA PSV in untreated hyperthyroid GD patients may reflect GD activity and thus MMI sensitivity. PMID- 17420112 TI - A mathematical approach to study combined effects of toxicants in vitro: evaluation of the Bliss independence criterion and the Loewe additivity model. AB - The study of interactions among toxicants is of fundamental interest and practical importance in toxicological sciences. However, a final agreement on the definition of agent interaction is lacking, and therefore, particular care should be adopted when using the terms additivity, synergism and antagonism, unless the exact toxicological pathways of the compounds studied are known. Two main different approaches, the Bliss independence criterion and the Loewe additivity model, have been generally used in co-exposure experiments. In some cases, they can present dramatically different results, depending on the slope of the pure dose-response curves of single substances. Here, we consider both models in in vitro experiments, where the dose-response curves can be extrapolated. Advantages and limitations of both approaches are discussed, using several mathematical simulations to better explain them, and applying the Hill function for the dose response model curve. Overall we conclude that the Loewe additivity model is slightly preferable because of a general higher biological plausibility. However, which model to use must be determined case by case and the evaluation can be aided by experimental approaches, such as the study of multiple biomarkers and asynchronous exposures. PMID- 17420113 TI - Medical utilization across the anxiety disorders. AB - Individuals with panic disorder often seek medical care for their symptoms prior to receiving effective treatment. However, little is known about how often, and in what settings, patients with other anxiety disorders present for medical treatment. In the present study, utilization of general and specialty medical services was coded via electronic chart review for 171 consecutive outpatients referred to an anxiety disorders clinic. Results indicated that panic disorder patients accrued the most medical visits overall, as well as the most frequent visits to cardiology, family medicine, and emergency medicine. Few differences in medical utilization were evident among patients with generalized anxiety disorder, obsessive-compulsive disorder, social phobia, and specific phobias. Patients with anxiety disorders appear to be frequent utilizers of medical services prior to receiving effective treatment. Our findings highlight the need for improved recognition and treatment of anxiety disorders, particularly panic disorder, in a number of medical settings. PMID- 17420114 TI - Contriving establishing operations: responses of individuals with developmental disabilities during a learning task. AB - The field of applied behavior analysis has utilized the ability to capture and contrive establishing operations in treating aberrant behavior in individuals with developmental disabilities. However, research on the use of establishing operations in the teaching of appropriate behavior is not as systematic. This study examined the effects of establishing operations on the responses of three males with mild to moderate intellectual disability during an incremental repeated acquisition procedure. Individuals responded more frequently and accurately during periods of longer deprivation (1 day and 2-3 days) than during shorter periods (15 min and 2h). These results have implications for conducting preference assessments, scheduling daily activities, maximizing responding and teaching new skills. PMID- 17420115 TI - Re-analysis of the risks attributed to ciprofloxacin-resistant Campylobacter jejuni infections. AB - Recent publications were compared and analysed in addition to novel surveillance data to assess the hypothesis that fluoroquinolone-resistant Campylobacter infections are causing more severe disease than susceptible infections. The available data did not support this hypothesis. There was no significant difference in duration of disease between susceptible and resistant infections. However, both for resistant and susceptible infections, disease symptoms were prolonged by on average 1-2 days for Campylobacter cases acquired during foreign travel. Nevertheless, the likelihood and duration of hospitalisation were not increased for cases related to foreign travel. These observations were confirmed by a new analysis of almost 11,000 cases. We conclude that fluoroquinolone resistant Campylobacter infections are not more severe than susceptible infections. PMID- 17420116 TI - Novel drug delivery system of hollow mesoporous silica nanocapsules with thin shells: preparation and fluorescein isothiocyanate (FITC) release kinetics. AB - Core-shell nanoparticles of Au@silica with a diameter of approximate 45-60 nm and wall thickness in range of 3-10 nm were synthesized by using 40 and 50 nm gold nanoparticles as the templates. The mesoporous particles are regulated by 3 aminopropyltrimethoxysilane addition. Hollow mesoporous silica nanocapsules (HMSNs) were prepared by using sodium cyanide to dissolve the gold cores. The characterization of Au@silica and HMSNs by transmission electronic microscope indicated that the silica shells were uniform and smooth, and also the porosity was proved by fluorescein isothiocyanate (FITC) release experiments. The ratio of hollow core to HMSNs is more than 70%. HMSNs were subsequently used as drug carrier to investigate FITC (as a model drug) release behaviors in vitro. Fluorescent spectrometry was performed to determine the release kinetics from the HMSNs. The release profiles are significantly different as compared with the control (free FITC), which show that HMSNs are good drug carriers to control drug release, and have high potential in therapeutic drugs delivery in future applications. PMID- 17420117 TI - Towards an intelligent medical system for the aesthetic evaluation of breast cancer conservative treatment. AB - OBJECTIVE: This work presents a novel approach for the automated prediction of the aesthetic result of breast cancer conservative treatment (BCCT). Cosmetic assessment plays a major role in the study of BCCT. Objective assessment methods are being preferred to overcome the drawbacks of subjective evaluation. METHODOLOGY: The problem is addressed as a pattern recognition task. A dataset of images of patients was classified in four classes (excellent, good, fair, poor) by a panel of international experts, providing a gold standard classification. As possible types of objective features we considered those already identified by domain experts as relevant to the aesthetic evaluation of the surgical procedure, namely those assessing breast asymmetry, skin colour difference and scar visibility. A classifier based on support vector machines was developed from objective features extracted from the reference dataset. RESULTS: A correct classification rate of about 70% was obtained when categorizing a set of unseen images into the aforementioned four classes. This accuracy is comparable with the result of the best evaluator from the panel of experts. CONCLUSION: The results obtained are rather encouraging and the developed tool could be very helpful in assuring objective assessment of the aesthetic outcome of BCCT. PMID- 17420118 TI - Non-traditional markers of atherosclerosis potentiate the risk of coronary heart disease in patients with type 2 diabetes and metabolic syndrome. AB - BACKGROUND AND AIMS: The aims of this study were to establish the prevalence of metabolic syndrome (MS), in type 2 diabetes mellitus (DM), according to National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) criteria, and to assess the association of MS with other cardiovascular (CV) risk factors in these patients. METHODS AND RESULTS: A cross-sectional study was conducted in 1610 patients with type 2 DM. Glycated hemoglobin A1c (HbA1c), total cholesterol, low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C), uric acid, fibrinogen, creatinine, and albumin/creatinine ratios were measured. The risk of coronary heart disease (CHD) was calculated using the UKPDS Risk Engine. Seventy percent of the diabetic population met the criteria for MS; central obesity and hypertension were the most common criteria. Subjects with MS had higher levels of HbA1c, LDL-C, non-HDL-C, uric acid, and fibrinogen compared to patients without MS. Similarly, microalbuminuria and a high triglyceride (Tg)/HDL-C ratio (a marker of small LDL-C) occurred more frequently in patients with MS. When patients with no history of CHD events were considered, mean CHD risk was greater in those with, than those without, MS. CONCLUSIONS: MS is highly prevalent in type 2 DM and is commonly associated with non-traditional CV risk factors. The diagnosis of MS seems to confer additional CHD risk in patients with type 2 diabetes. PMID- 17420119 TI - Local development of effector and memory T helper cells. AB - Clonal evolution underpins all facets of adaptive immunity. In particular, antigen-specific helper T (Th) cell development is central to high-affinity B cell immunity and protective vaccination. Dendritic cell maturation and TCR affinity-based selection mechanisms control the recruitment and effective propagation of preferred antigen-specific Th cell cohorts in local lymphoid tissue. Importantly, follicular B helper T (T(FH)) cells emerge as the specialized local effector Th cells that orchestrate the stepwise development of B cell immunity in these local environments. Recent studies also introduce the role of persistent antigen in the development of effector Th cells with evidence for long-term antigen depots that might contribute to local antigen-specific Th cell memory. PMID- 17420120 TI - Concentration dependence of IgG-protein A affinity studied by wireless electrodeless QCM. AB - The binding affinity between human immunoglobulin G (IgG) and protein A was studied by the homebuilt wireless-electrodeless quartz crystal microbalance (QCM). Protein A was immobilized on the electrodeless AT-cut quartz plate of 0.05 mm thick and its fundamental resonance frequency near 34 MHz was measured by a noncontacting manner using a line antenna. The vibrational analysis was performed to ensure higher sensitivity of the electrodeless QCM. A flow-cell system was fabricated to continuously measure the resonance frequency during the injection sequence of the IgG solutions with concentrations of 1-20,000 ng/mL. The exponential frequency changes were recorded to determine the affinity based on the Langmuir kinetics. The equilibrium constant K(A) significantly varied between 6 x 10(6) and 6 x 10(10) M(-1), depending on the IgG concentration, which is attributed to various formations of IgG-protein A complexes. PMID- 17420121 TI - Fermentation of molasses by Zymomonas mobilis: effects of temperature and sugar concentration on ethanol production. AB - Fermentations utilizing strains of Zymomonas mobilis, in place of the traditional yeasts, have been proposed due their ethanol yields being close to theoretical. Ethanol production from sugar cane molasses was analyzed under different culture conditions using Z. mobilis in batch fermentation. The total reducing sugars (TRS) concentrations in the molasses, temperature, agitation and culture time effects were studied simultaneously through factorial design. The best conditions for ethanol production were 200 g L(-1) of total reducing sugars in the molasses, temperature of 30 degrees C and static culture and time of fermentation of 48 h, achieving 55.8 g L(-1). The pH of the medium was kept constant during the experiments, showing that molasses presents a buffering effect. PMID- 17420122 TI - Selective and dual action orally active inhibitors of thrombin and factor Xa. AB - The synthetic entry to new classes of dual fXa/thrombin and selective thrombin inhibitors with significant oral bioavailability is described. This was achieved through minor modifications to the sulfonamide group in our potent and selective fXa inhibitor (E)-2-(5-chlorothien-2-yl)-N-{(3S)-1-[(1S)-1-methyl-2-(morpholin-4 yl)-2-oxoethyl]-2-oxopyrrolidin-3-yl}ethenesulfonamide and these observed activity changes have been rationalised using structural studies. PMID- 17420123 TI - Synthesis and structure-activity relationships of 4-hydroxy-4-phenylpiperidines as nociceptin receptor ligands: Part 2. AB - A series of 4-[2-(aminomethyl)phenyl]-1-[bis(2-chlorophenyl)methyl]-4 hydroxypiperidine analogs has been identified as nociceptin receptor ligands. These compounds display high affinity and functional activity at the nociceptin receptor. The synthesis and structure-activity relationships at the C-4 phenyl and N-1 positions are described and the antitussive activity of a selected compound is reported. PMID- 17420124 TI - Concise synthesis of dideoxy-epigallocatechin gallate (DO-EGCG) and evaluation of its anti-influenza virus activity. AB - Dideoxy-epigallocatechin gallate (DO-EGCG) (2), a simplified analog of naturally occurring EGCG (1), was efficiently prepared by directly introducing a ketone group at C3 and successive reduction to the sec-alcohol with 2,3-cis stereochemistry. Compound 2 showed potent anti-influenza virus activity, indicating that the hydroxyl substituents on the A-ring are not crucial for anti influenza virus activity. PMID- 17420125 TI - Penicillin-bound polyacrylate nanoparticles: restoring the activity of beta lactam antibiotics against MRSA. AB - This report describes the preparation of antibacterially active emulsified polyacrylate nanoparticles in which a penicillin antibiotic is covalently conjugated onto the polymeric framework. These nanoparticles were prepared in water by emulsion polymerization of an acrylated penicillin analogue pre dissolved in a 7:3 (w:w) mixture of butyl acrylate and styrene in the presence of sodium dodecyl sulfate (surfactant) and potassium persulfate (radical initiator). Dynamic light scattering analysis and atomic force microscopy images show that the emulsions contain nanoparticles of approximately 40 nm in diameter. The nanoparticles have equipotent in vitro antibacterial properties against methicillin-susceptible and methicillin-resistant forms of Staphylococcus aureus and indefinite stability toward beta-lactamase. PMID- 17420126 TI - Bidirectional Eph-ephrin signaling during axon guidance. AB - Ephrins are cell-surface tethered guidance cues that bind to Eph receptor tyrosine kinases in trans on opposing cells. In the developing nervous system, the Eph-ephrin signaling system controls a large variety of cellular responses including contact-mediated attraction or repulsion, adhesion or de-adhesion, and migration. Eph-ephrin signaling can be bidirectional, and is subject to modulation by ectodomain cleavage of ephrins and by Eph-ephrin endocytosis. Recent work has highlighted the importance of higher-order clustering of functional Eph-ephrin complexes and the requirement for Rho GTPases as signal transducers. Co-expression of Ephs and ephrins within the same cellular membrane can result in Eph-ephrin cis interaction or in lateral segregation into distinct domains from where they signal opposing effects on the axon. PMID- 17420129 TI - Clinical research 3: Sample selection. AB - This research series is aimed at clinicians who wish to develop research skills, or who have a particular clinical problem that they think could be addressed through research. The series aims to provide insight into the decisions that researchers make in the course of their work and to also provide a foundation for decisions that nurses may make in applying the findings of a study to practice in their own Unit or Department. The series emphasises the practical issues encountered when undertaking research in critical care settings: readers are encouraged to source research methodology textbooks for more detailed guidance on specific aspects of the research process. PMID- 17420130 TI - Pituitary abscess in a pregnant woman. AB - Pituitary abscess is a rare and potentially lethal condition. Pituitary abscess in a pregnant woman has not been previously described. A 38-year-old pregnant woman (34 weeks gestation) with a pituitary mass complained of a progressive headache and sudden visual impairment. She was afebrile and had no inflammatory symptoms on admission. On MRI, the preoperative diagnosis was pituitary adenoma with sphenoid sinusitis. She underwent an uncomplicated transsphenoidal procedure for removal of the pituitary mass. The next day, labor commenced and a healthy preterm baby was delivered. Pathologic examination of the intrasellar mass showed polymorphonuclear cells, debris and no tumor cells. The sellar contents were cultured and Streptococcus viridans was grown. To our knowledge this is the first case of pituitary abscess reported during pregnancy. Although the patient was pregnant, the transsphenoidal approach was safe for the mother and the fetus. Surgical drainage and antibiotic therapy are required for the definitive treatment of this condition. PMID- 17420131 TI - Calculation of binding energy using BLYP/MM for the HIV-1 integrase complexed with the S-1360 and two analogues. AB - Integrase (IN) is one of the three human immunodeficiency virus type 1 (HIV-1) enzymes essential for effective viral replication. S-1360 is a potent and selective inhibitor of HIV-1 IN. In this work, we have carried out molecular dynamics (MD) simulations using a hybrid Quantum Mechanics/Molecular Mechanics (QM/MM) approach, to determine the protein-ligand interaction energy for S-1360 and two analogues. Analysis of the MD trajectories reveals that the strongest protein-inhibitor interactions, observed in the three studied complexes, are established with Lys-159 residue and Mg(2+) cation. Calculations of binding energy using BLYP/MM level of theory reveal that there is a direct relationship between this theoretical computed property and the experimental determined anti HIV activity. PMID- 17420132 TI - Molecular modeling study for the binding of zonisamide and topiramate to the human mitochondrial carbonic anhydrase isoform VA. AB - Zonisamide and topiramate are two antiepileptic drugs known to induce weight loss in epilepsy patients. These molecules were recently shown to act as carbonic anhydrase (CA) inhibitors, being presumed that the weight loss may be due to the inhibition of the mitochondrial isozymes CA VA and CA VB involved in metabolic processes, among which lipid biosynthesis. To better understand the interaction of these compounds with CAs, here, we report a homology modeling and molecular dynamics simulations study on their adducts with human carbonic anhydrase VA (hCA VA). According to our results, in both cases the inhibitor sulfamate/sulfonamide moiety participates in the canonical interactions with the catalytic zinc ion, whereas the organic scaffold establishes a large number of van der Waals and polar interactions with the active site cleft. A structural comparison of these complexes with the corresponding homologues with human carbonic anhydrase II (hCA II) provides a rationale to the different affinities measured for these drugs toward hCA VA and hCA II. In particular, our data suggest that a narrower active site cleft, together with a different hydrogen bond network arrangement of hCA VA compared to hCA II, may account for the different Kd values of zonisamide and topiramate toward these physiologically relevant hCA isoforms. These results provide useful insights for future design of more isozyme-selective hCA inhibitors with potential use as anti-obesity drugs possessing a novel mechanism of action. PMID- 17420133 TI - Novel cationic lipophilic peptides for oligodeoxynucleotide delivery. AB - In search of new oligodeoxynucleotide (ODN) delivery agents, we evaluated novel peptides derived from core peptide H-GLRILLLKV-OH (CP). CP is a fragment designed from the T-cell antigen receptor (TCR) alpha-chain transmembrane sequence. CP was able to enter cells including T-cells and inhibited interleukin-2 (IL-2) production. To examine the effect of increased lipophilicity on cellular uptake and activity of CP, a lipoamino acid (2-aminododecanoic acid) was incorporated into peptide CP resulting in 2-aminodecanoyl-CP (LP). The toxicity of CP and LP was assessed by measuring the haemolytic activity. Neither compound caused any haemolysis of red blood cells. We have also compared the biological activities of the CP and LP. Using a T-cell antigen presentation assay, the more lipophilic LP caused greater inhibition of IL-2 production than the parent CP in the antigen stimulated T-cells. The LP also showed increased permeability than CP in the Caco 2 cell assay. We utilised the enhanced cell permeability property of LP in oligodeoxynucleotide ODN1 delivery. Isothermal titration calorimetry (ITC) suggested that CP and LP complex with ODN1 in a 12:1 (CP:ODN1) and 15:1 (LP:ODN1) ratio. These complexes were then transfected into human retinal pigment epithelial cells. The level of transfection was measured by the decreased production of the protein human vascular endothelial growth factor (hVEGF). The results revealed greater transfection efficiency for both CP and LP (47%, 55% more inhibition) compared to commercially available transfection agent cytofectin GSV. These results suggested that the CP and particularly its lipophilic analogue LP have the potential to be used as oligodeoxynucleotide delivery systems. PMID- 17420134 TI - Interpreting MADCAP: parallelism not divergence. PMID- 17420135 TI - Use of a biophysical model of atrial fibrillation in the interpretation of the outcome of surgical ablation procedures. AB - OBJECTIVE: To determine the adequacy of 'in silico' biophysical models of atrial fibrillation (AF) in the design of different ablation line patterns. BACKGROUND: Permanent AF is a severe medical problem for which (surgical) ablation is a possible treatment. The ideal ablation pattern remains to be defined. METHODS: Forty-six consecutive adult patients with symptomatic permanent drug refractory AF underwent mitral surgery combined with non-transmural, (n=20) and transmural (n=26) radiofrequency Minimaze. The fraction of 'in vivo' conversions to sinus rhythm (SR) in both groups was compared with the performance of the fraction of 'in silico' conversions observed in a biophysical model of permanent AF. The simulations allowed us to study the effectiveness of incomplete and complete ablation patterns. A simulated, complete, transmural Maze III ablation pattern was applied to 118 different episodes of simulated AF set-up in the model and its effectiveness was compared with the clinical results reported by Cox. RESULTS: The fraction of conversions to SR was 92% 'in vivo' and 88% 'in silico' (p=ns) for transmural/complete ablations, 60% respectively 65% for non transmural/incomplete Minimaze (p=ns) and 98% respectively 100% for Maze III ablations (p=ns). The fraction of conversions to SR 'in silico' correlated with the rates 'in vivo' (r2=0.973). CONCLUSIONS: The fraction of conversions to SR observed in the model closely corresponded to the conversion rate to SR post surgery. This suggests that the model provides an additional, non-invasive tool for optimizing ablation line patterns for treating permanent AF. PMID- 17420137 TI - Surgical treatment of a pulmonary artery aneurysm. AB - We would like to report a rare case of post-stenotic aneurysm of the pulmonary trunk and its left branch in a 51-year-old man. His cardiac disease, which was first diagnosed at the age of 4, was left untreated because of absence of symptoms and normal physical development. A CT scan, recently performed because of decrease in exercise tolerance and worsening dyspnea, showed a pulmonary artery aneurysm (52 mm x 79 mm). The echocardiography revealed a severe pulmonary commissural stenosis. Through a surgical approach the pulmonary trunk and its left branch were excised and reconstructed using a 30 mm Dacron graft; the right pulmonary branch was then reimplanted on the right side of the tube. The patient's postoperative course was uneventful. He was discharged on the seventh postoperative day and there were no adverse events or complications at 1- and 3 month follow-up. PMID- 17420138 TI - Some reflections on talc poudrage. PMID- 17420139 TI - Spontaneous pneumomediastinum: 41 cases. AB - OBJECTIVE: Spontaneous pneumomediastinum is characterized by the presence of interstitial air in the mediastinum without any apparent precipitating factor. The purpose of this study is to review and discuss our experience with this condition. METHODS: A descriptive, retrospective study of 41 cases--34 men (83%) and 7 women (17%)--treated at our hospital for spontaneous pneumomediastinum from January 1990 through June 2006. RESULTS: The mean age of the patients was 21 years (range, 14-35 years). Notably, 22% of patients had a prior history of asthma. No precipitating factor was identified in 51% of cases while onset was associated with physical effort in 12%. Chest pain (85%) and dyspnea (49%) were the most common symptoms. Subcutaneous emphysema, which presented in 71% of patients, was the most common sign. Pneumomediastinum was diagnosed by plain chest radiography in all cases. In certain cases, a computed tomography scan of the chest, contrast-enhanced swallow, or bronchoscopy was performed. All patients were admitted to the hospital with good progress and no instances of morbidity or mortality. Treatment included analgesia, rest, and/or initial oxygen therapy. The mean length of hospital stay was 5 days (range, 1-9 days) with only one case of early recurrence, which was resolved satisfactorily. CONCLUSIONS: Spontaneous pneumomediastinum is a benign process primarily affecting young men. Despite its low incidence, spontaneous pneumomediastinum should be considered in the differential diagnosis of acute chest pain because it requires a high index of suspicion. Patients with spontaneous pneumomediastinum respond well to medical treatment, with no recurrence in the great majority of cases. PMID- 17420140 TI - Kidney and splanchnic handling of interleukin-6 in humans. AB - Chronic elevation of circulating Interleukin-6 (IL-6) is observed in elderly individuals as well as in several illnesses, including chronic kidney diseases. A number of cells and tissues possess the ability to metabolize significant amounts of IL-6 in vitro. However, information on signals and mechanisms by which IL-6 is removed from blood in humans is still incomplete. To assess the individual role of splanchnic organs and kidney on IL-6 inter-organ exchange we used the IL-6 mass-balance technique across the hepato-splanchnic bed and kidney in six subjects with normal renal and liver function undergoing diagnostic venous catheterizations. Both in the hepatic and renal veins IL-6 levels were significantly lower (p=0.041 and 0.038, respectively), than in the artery. The fractional extraction of IL-6, i.e., the percentage of arterial IL-6 extracted after a single pass, was greater across the splanchnic organs (18%) than across the kidney (8%). Accordingly, IL-6 plasma clearance across splanchnic organs was greater than across the kidney and the sum of kidney and splanchnic removal accounted for as much as 63% of the estimated adipocyte IL-6 release. Our data demonstrate that, although the individual contribution to removal is different, both splanchnic organs and kidneys affect in a significant way the disposal of IL 6 in humans. According, both liver and kidney dysfunction could affect the handling of this proinflammatory cytokine and favour a chronic inflammatory response. PMID- 17420141 TI - Comparison of two within-household selection methods in a telephone survey of substance abuse and dependence. AB - PURPOSE: Random-digit dial telephone surveys often rely on the random selection of one respondent within the household. We compared a new method of within household selection to a standard "next birthday" approach on selected survey process measures, respondent characteristics, and substantive results. METHODS: From October 2004 through June 2005, we conducted a survey of adults in Minnesota to obtain information about substance use and need for treatment. Control group respondents (n = 1944) were selected using the "next birthday" method, and experimental group respondents (n = 1086) were selected using a new method developed by Rizzo, Brick, and Park. We assessed group differences for survey process measures, such as the number of attempts to interview and the refusal, response, and cooperation rates. We also examined whether the groups differed in demographic factors, substance use, and mental health. RESULTS: The experimental group had higher rates of refusal and lower response and cooperation rates. Demographic factors and most measures of substance use and mental health did not differ significantly between groups. CONCLUSIONS: The experimental method of within-household selection developed by Rizzo and colleagues does not offer advantages over the classic "next birthday" method. Study limitations are discussed and opportunities for future research are identified. PMID- 17420142 TI - Risk-adjusted mortality rates of elderly veterans with hip fractures. AB - PURPOSE: The goal of this research was to estimate 12-month survival rates for a large sample of elderly veterans after hip fracture with a risk-adjusted model and to compare the results of men to those of women. METHODS: The study design was a retrospective, secondary data analysis of national Veterans Health Administration (VHA) Medicare beneficiaries. The study population was 43,165 veterans with hip fracture first admitted to a Medicare-eligible facility during our specified enrollment period of 1999-2002. Measurement was a Cox proportional hazard model or survival analysis of hip fracture patients with an outcome of death over a 1 year period after discharge controlled by age, gender, and selected Elixhauser comorbidities. RESULTS: The unadjusted, 1 year mortality rates (30 days = 9.7%, 90 days = 17.5%, 180 days = 24%, 365 days = 32.2%) were slightly higher than the adjusted rates (30 days = 8.9%, 90 days = 15.6%, 180 days = 21.8%, 1 year = 29.9%). The mortality odds for women 12 months after hip fracture were 18%, compared with 32% for men. The comorbidity adjustment suggested that the presence of metastatic cancer increased the risk of death by almost 4 times compared with those patients without this diagnosis. Other particularly high-risk conditions included congestive heart failure, renal failure, liver disease, lymphoma, and weight loss, each of which increased the 1 year mortality risk by approximately two-fold. CONCLUSIONS: One in 3 elderly male veterans who sustain a hip fracture dies within 1 year. Our work represents the first large study of hip fractures with a predominantly male sample and confirms that men have a higher mortality risk than women, as reported by previous researchers who used smaller samples that were mostly female. Fracture patients with metastatic cancer, renal failure, lymphoma, weight loss, and liver disease have higher mortality risks. The adverse outcomes associated with hip fracture argue for clinical intervention strategies, such as gait and balance testing, and osteoporosis diagnosis that may prevent fractures in both genders. PMID- 17420143 TI - Electrophoretic and Western blot analyses of the lipopolysaccharide and glycocalyx of Flavobacterium psychrophilum. AB - Flavobacterium psychrophilum is the aetiological agent of bacterial coldwater disease (CWD) and rainbow trout fry syndrome (RTFS) and it has emerged as one of the most significant bacterial pathogens in salmonid aquaculture worldwide. Previous studies have suggested that the O-polysaccharide (O-PS) component of the lipopolysaccharide (LPS) of F. psychrophilum is highly immunogenic and may be involved in eliciting a protective immune response in rainbow trout (Oncorhynchus mykiss Walbaum). In the present study, SDS-PAGE and Western blotting techniques were used to analyse the carbohydrate antigens of F. psychrophilum. Our analysis identified two distinct carbohydrate-banding patterns. One banding pattern corresponds with LPS, and we hypothesise that the other carbohydrate-banding pattern is that of the loosely associated glycocalyx of F. psychrophilum. Electron microscopy of F. psychrophilum cells immunogold labelled with a monoclonal antibody specific for this banding pattern supports this hypothesis as the outermost layer of the bacterium was heavily labelled. This is a significant finding because the immunogenic antigens that have been referred to as the O-PS of LPS, and implicated as potential vaccine candidate antigens, appear to be components of the glycocalyx of F. psychrophilum. This research suggests that the glycocalyx of F. psychrophilum may be an important antigen to consider for the development of a vaccine to control CWD and RTFS. PMID- 17420145 TI - Clinical use of antiepileptic drugs at a referral centre for epilepsy. AB - PURPOSE: The National Centre for Epilepsy in Norway admits patients with refractory epilepsy from the whole country. The purpose of this study was to investigate how antiepileptic drugs (AEDs) are used at the centre and compare it with the total consumption in the country and international guidelines regarding clinical use of AEDs. MATERIAL AND METHODS: A prevalence study was carried out from patient records from 264 patients (136 adults and 128 children). The use of AEDs, gender, polytherapy, common drug combinations, serum concentration measurements, concomitant medication and comorbid conditions were investigated. RESULTS: Of the 15 AEDs in use, valproate, lamotrigine and levetiracetam were most frequently used. In the country at large, carbamazepine, valproate and lamotrigine were used the most. Valproate and lamotrigine occurred most frequently in combination. In adults, oxcarbazepine and topiramate were used more frequently in women than in men. Children used benzodiazepines three times as often as adults. Newer AEDs were mostly used for partial seizures, in accordance with international guidelines. Thirty-five percent of adults and 20% of children suffered from comorbid CNS-related conditions. The use of concomitant medication was widespread. Serum concentrations were in accordance to recommended therapeutic ranges. CONCLUSION: The results demonstrate that newer AEDs were much more frequently used at the epilepsy centre than in the country as a whole. The use of polytherapy was common, and therapeutic drug monitoring was used for optimal individualized drug treatment. AEDs were used in accordance to international guidelines and recommendations, which can serve as a reference for other prescribers. PMID- 17420144 TI - DNA-based taxonomy for associating adults and larvae in multi-species assemblages of chafers (Coleoptera: Scarabaeidae). AB - DNA sequences provide a universal character system in taxonomy for associating all developmental stages of organisms, but ambiguity arises due to genetic variation within species. The problem is compounded where target groups are less well studied or incompletely represented in DNA databases. Here we investigate the utility of DNA for larval-adult species associations within chafer (Coleoptera: Scarabaeidae) communities from four sites in the tropical lowlands of Nepal. We sequenced ca. 1600 bp of mitochondrial cox1 and rrnL and 700 bp of nuclear 28S rRNA from 250 larval and adult specimens. Individuals were grouped into putative species using statistical parsimony analysis and population aggregation analysis (PAA), whereby specimens from each locality were grouped according to the presence of diagnostic nucleotides. In addition, species membership was determined based on shifts in branching rates on clock-constrained trees to detect the putative transition from speciation to population coalescence patterns. Using these two methods we delineated between 48 and 56 groups, of which 16-20 were composed of larval and adult individuals. Nuclear and mtDNA based groups were highly congruent; variation of 28S rRNA within groups was very low, while one widespread 28S rRNA genotype was universally found in a paraphyletic group of five mtDNA clusters. Linnean names could be assigned to 19 groups, and hence between 86.1% and 92.7% of larval specimens could be associated to species by their membership in clearly delineated groups that contained fully identified adults. The remaining larvae were delineated as five species, four of which could be assigned to Anomala or Adoretus based on their phylogenetic position. We conclude that the sequence variation was highly structured in this complex assemblage of chafers and that any given individual (larva or adult) can be readily associated with a particular DNA group using the criterion of diagnos ability. The association of different developmental stages therefore becomes a matter of determining the extent of the DNA-based groups, rather than matching of sequences from adult and larval individuals. This indicates the need for a purely sequence-based taxonomic system when associating different life stages via DNA. PMID- 17420146 TI - Novel 3'-phosphoadenosine-5'-phosphatases from extremely halotolerant Hortaea werneckii reveal insight into molecular determinants of salt tolerance of black yeasts. AB - The 3'-phosphoadenosine-5'-phosphatase encoded by HAL2 gene, is a ubiquitous enzyme required for the removal of the cytotoxic 3'-phosphoadenosine-5'-phosphate produced during sulfur assimilation in eukaryotes. Salt toxicity in yeast and plants results from Hal2 inhibition by sodium or lithium ions. Two novel HAL2 like genes, HwHAL2A and HwHAL2B, have been cloned from saltern-inhabited extremely halotolerant black yeast Hortaea werneckii. Expression of both HwHAL2 isoforms was differentially inducible upon salt. When the HwHAL2 genes were transferred from such a halotolerant species into the salt sensitive Saccharomyces cerevisiae, the resulting organism can tolerate 1.8M NaCl or 0.8M LiCl, the highest reported salt concentrations at which S. cerevisiae can grow. With genetic and biochemical validation we demonstrated the critical HwHal2B sequence motif--the META sequence--common only to Dothideales fungi, with evident effect on the HwHal2B-dependent salt tolerance. These results may have significance for biosaline agriculture in coastal environments. PMID- 17420147 TI - Fe-S cluster proteins are intracellular targets for nitric oxide generated luminally at the gastro-oesophageal junction. AB - In human, high concentrations of nitric oxide are generated at the gastro oesophageal junction through entero-salivary recirculation of dietary nitrate. Nitric oxide is known to have a high affinity for Fe-S cluster proteins. The aim of this study is to investigate whether nitric oxide arising from the lumen diffuses into the adjacent tissue where it can interact with Fe-S proteins both in a rat animal model and human. An electron paramagnetic resonance detectable complex, dinitrosyl dithiolato iron complex (DNIC), was used as a biomarker for the interaction between Fe-S proteins and nitric oxide. The generation of the complex was evaluated in resected gastric tissue of nitrite-administered rat or biopsy specimens from human after nitrate ingestion. The activity of aconitase, one of the Fe-S cluster proteins, was also determined. The signal of the complex was observed at the rat gastro-oesophageal junction where luminal generation of nitric oxide from nitrite was maximal, and the intensity increased in a dose- and time-dependent manner. The appearance of the complex was accompanied by a significant inhibition of the aconitase activity at that site. The complex appeared in biopsy specimens from the gastro-oesophageal junction in three of five men after nitrate ingestion. Since DNIC is considered to be a decomposition product when Fe-S cluster proteins interact with nitric oxide, the appearance of the signal provides direct evidence that nitric oxide arising from the lumen can destroy such proteins. DNIC formation may represent the cellular mechanism responsible for the high prevalence of disease at the gastro-oesophageal junction. PMID- 17420148 TI - Epidemiology of the acute abdominal crisis: will it challenge current knowledge? PMID- 17420149 TI - A review of viral diseases of the European wild boar: effects of population dynamics and reservoir role. AB - There has been a worldwide increase in the number and geographical spread of wild boar populations in recent decades leading to an increase in both the circulation of disease agents and greater contact with domestic animals and humans. Diseases affect the population dynamics of wildlife but the effects of most viral diseases on the European wild boar are largely unknown. Many viral diseases present in domestic pig populations are also present in wild boars where they can provide a disease reservoir, as is clearly the case with classical swine fever, but little is known about other viral diseases such as porcine circovirus diseases or hepatitis E. This review considers the current scientific knowledge of the effects of viral diseases on wild boar populations and their role as potential disease reservoirs. The focus is on those viral diseases of domestic swine and wild boars that are included as notifiable by the Office International des Epizooties (OIE). PMID- 17420150 TI - Long-term follow-up of bone mineral density in childhood hypophosphatasia. AB - OBJECTIVE: Hypophosphatasia (HP; MIM 241510) is an inborn error of bone metabolism, characterized by a genetic defect in the gene of the tissue-non specific alkaline phosphatase TNSALP. Long-term data on bone mineral density measurements are not available. METHODS: We have analyzed changes of bone mineral density (pQCT and DXA) prospectively during 4years of follow-up in a cohort of 6 patients with childhood HP. RESULTS: At diagnosis hypermineralization of the trabecular bone in the metaphyseal area of long bones in affected children was noted. During 4 years of follow-up a gradual, significant decrease of mineralization was noted in the radial metaphyses. In contrast, BMC by DXA and total body DXA values were stable in comparison to healthy controls. During follow-up a systemic hyperprostaglandinism was documented in the majority of the patients. Non-steroidal anti-inflammatory drug treatment was evaluated by measuring prostaglandin excretion in the urine. CONCLUSIONS: Metaphyseal hypermineralization in childhood HP, which might be a compensation for a mechanically incompetent bony structure, decreased over time. There might be a pathophysiological link to continually elevated systemic prostaglandins. PMID- 17420151 TI - Neurotrophic ulcer after extra-capsular cataract operation. AB - Although neurotrophic ulcers due to herpes zoster are seldom, there may be challenging cases. Especially neurotrophic corneal ulcers after cataract operations should arise the possibility of a previous herpes zoster attack and the treatment plan should be prepared accordingly. This case highlights the importance of thorough evaluation of cataract patients in order not to miss a previous diagnosis of herpes. PMID- 17420152 TI - A 12-month prospective clinical trial of comfilcon A silicone-hydrogel contact lenses worn on a 30-day continuous wear basis. AB - PURPOSE: To evaluate the clinical performance of comfilcon A (Biofinity) during 12 months of continuous wear compared to other silicone-hydrogel lenses. METHOD: Forty-five subjects were fitted in one eye with the comfilcon A (test) lens. For 22 subjects, the other eye was fitted with lotrafilcon A (group A) and for the remaining 23 subjects the other eye was fitted with balafilcon A (group B). Twelve-month data are presented on 48 measured variables. RESULTS: There were no serious adverse events during the course of the study. A total of 33 subjects completed the study, with only 4 discontinuations for lens-related reasons, spread proportionately between the lenses. On preference scales, the test lens was superior to both control lenses for comfort (p<0.05) and overall preference (p<0.05) and in group A for vision (p<0.05). On grading scales, the test lens scored better than both control lenses for overall comfort (p<0.1 for group A, p<0.05 for group B), comfort during the day (p<0.05), end-of-day comfort (p<0.05) and overall preference (p<0.05), and better in group A for general vision quality (p<0.01), night vision quality (p<0.01) and handling (p<0.05). All lenses performed to a comparable degree on the basis of microcysts and corneal staining. There were significant differences between lenses in producing limbal redness (p<0.05), bulbar conjunctival hyperemia (p<0.1) and conjunctival NaFl staining (p<0.01) with the control lens in group A showing the greatest effect and the control lens in group B the least. CONCLUSION: Comfilcon A offers performance advantages over first generation silicone-hydrogel materials for continuous wear. PMID- 17420153 TI - Theoretical investigations of zero-field splitting of excited states for 3d3 ions in trigonal crystal fields. AB - By taking into account slight interactions, i.e. spin-spin, spin-other-orbit and orbit-orbit interactions, in addition to spin-orbit interaction, the zero-field splitting of ground state and low excited states and g factors of ZnGa2O4:Cr3+ crystal have been interpreted systematically. And the contributions to zero-field splitting arising from slight magnetic interaction and trigonal crystal field are investigated. It is found that there exist combined mechanism between magnetic interactions and trigonal crystal field. PMID- 17420154 TI - Circadian rhythm sleep disorders in the blind and their treatment with melatonin. AB - People who are blind, in addition to having to cope with partial or no sight, have an added handicap; the transmission of ocular light from the retina to their circadian clock is impaired. At its worse, for example in people with both eyes enucleated, this lesion results in desynchronisation of the biological clock (located in the hypothalamic suprachiasmatic nuclei) from the 24h day/night environment. While in a desynchronised state, symptoms akin to jet lag are experienced (e.g., daytime sleepiness, poor night sleep, reduced alertness and performance during waking). This is a lifelong condition. Daily administration of exogenous melatonin is the current treatment of choice for this so-called "non 24h sleep/wake disorder". Melatonin has been shown to correct the underlying circadian rhythm abnormality as well as improve sleep and reduce daytime napping. The effectiveness of melatonin therapy depends upon its time of administration relative to the timing of the person's circadian clock. If practicable, assessment of an individual's circadian phase (by measurement of the endogenous melatonin rhythm in plasma, saliva or urine) is recommended prior to commencing treatment to optimise melatonin's effectiveness. PMID- 17420155 TI - Pacemaker infections: a 10-year experience. AB - BACKGROUND: Infection is a major complication of pacemaker and defibrillator (PPM/ICD) implantation. The experience in an Australian regional centre is reported. METHODS: Ten years' (1994-2004) cases of PPM/ICD infection retrospectively analysed and compared to overall insertion data; management and outcomes examined. RESULTS: A total 39 cases (79.5% male, median age 71.3 years) identified, 24 in the primary centre where 1481 procedures performed (infection rate 1.6%). Patients with infection had average 2.2 procedures performed (odds ratio for infection if >1 procedure=4.7); 14 (36%) first implantations, 35 (90%) pacemakers, 11 (28%) recurrent. No difference in operation duration or difficulty between infected and non-infected cases. Infection in 18 cases (46%) involved lead/s, 16 (41%) generator and 5 (13%) both. Median time to presentation was 7.9 months. Echocardiography demonstrated lead vegetations in 8 cases. Organisms identified in 25 (64%)-92% Staphylococci (65% S. aureus); blood cultures positive in 18. PPM/ICD removed in 26 (67%), including lead/s in 89%; average hospital stay 37 days. One death attributable to PPM/ICD infection (mortality 2.6%), median follow-up 29.3 months. CONCLUSIONS: A PPM/ICD infection rate of 1.6% (endocarditis 0.3%) demonstrated. Second and subsequent procedures carried almost 5 times greater infection risk. PMID- 17420156 TI - Myocardial release of nitric oxide during ischaemia and reperfusion: effects of L arginine and hypercholesterolaemia. AB - AIMS: Nitric oxide (NO) may modulate myocardial ischaemia/reperfusion (I/R) injury, but effects of hypercholesterolaemia on myocardial NO release during I/R are unknown. METHODS: A NO-specific carbon fibre electrode continuously measured coronary sinus [NO] during 60 min low-flow ischaemia (1 ml/min) and 60 min free reperfusion (I/R) in isolated rabbit hearts. Experimental groups (n=7 per group) were control, L-arginine supplement (200 microM), N-nitro-L-arginine methyl ester (L-NAME) treatment (8 microM) and hypercholesterolaemic. RESULTS: During early I, NO release decreased markedly in control (-1356+/-286 pmol/min/g) and L-arginine (-1972+/-172) groups, but less in L-NAME (-441+/-89) and hypercholesterolaemic ( 602+/-164) groups (both p<0.01 vs. controls). No increase in NO release during I was seen in any group. After R, NO release increased above baseline in control (+2333+/-591 pmol/min/g) and L-arginine (+1048+/-278) groups and hypercholesterolaemic (+1100+/-478) (p<0.05 vs. pre-ischaemia each group). There was little increase in NO release in the L-NAME group (+436+/-247 pmol/min/g, p<0.05 vs. controls). In each group, myocardial NO release declined towards pre ischaemic levels during 60 min R. Hearts treated with L-arginine had similar NO release but better functional recovery than controls (p<0.01). Treatment with L NAME was also associated with better functional recovery than in controls or hypercholesterolaemic hearts. CONCLUSION: Myocardial NO release declines rapidly during ischaemia, but increases above baseline during early reperfusion. Improved function after L-arginine treatment appears to be independent of effects upon NO release. Hypercholesterolaemia is associated with reduced myocardial NO release, under both baseline conditions and during ischaemia and reperfusion. PMID- 17420157 TI - Blood pressure and hypertension in an American colony (Puerto Rico) and on the USA mainland compared, 1886-1930. AB - We compare blood pressure and hypertension between adult men on the USA mainland and in Puerto Rico born during 1886-1930 to test hypotheses about the link between cardiovascular health and large socioeconomic and political changes in society: (a) 8853 men surveyed in Puerto Rico in 1965 and (b) 1449 non-Hispanic White men surveyed on the mainland during 1971-1975. Systolic and diastolic blood pressure and hypertension were regressed separately on demographic and socioeconomic variables and cardiovascular risk factors. Mainland men not taking anti-hypertensive medication showed statistically significant improvements in systolic blood pressure and hypertension at the beginning of the century and men in Puerto Rico showed improvements in diastolic blood pressure but only during the last two quinquenniums. An average man born on the mainland during the last birth quinquennium (1926-1930) had 7.4-8.7 mmHg lower systolic blood pressure and was 61% less likely to have systolic hypertension than one born before 1901. On average Puerto Rican men born during 1921-1925 had approximately 1.7 mmHg lower diastolic blood pressure than men born before 1901. Analyses of secular trends in cardiovascular health complements analyses of secular trends in anthropometric indicators and together provide a fuller view of the changing health status of a population. PMID- 17420158 TI - Transjugular intrahepatic portosystemic shunt: a case report of rescue management of unrestrainable variceal bleeding in a pregnant woman. AB - Liver cirrhosis complications in pregnant women are frequent and death rate secondary to variceal bleeding is relevant. Both sclerotherapy and banding ligation seem to be safe procedures in pregnancy; when bleeding is not arrested endoscopically an emergency transjugular intrahepatic portosystemic shunt should be considered, but data regarding pregnant cirrhotic women are scarce. We describe the case of a pregnant woman at 14 weeks of gestation who underwent management of acute variceal bleeding by transjugular intrahepatic portosystemic shunt. Transjugular intrahepatic portosystemic shunt may represent a rescue treatment for failed attempts of band ligation or sclerotherapy. PMID- 17420159 TI - Peppermint oil (Mintoil) in the treatment of irritable bowel syndrome: a prospective double blind placebo-controlled randomized trial. AB - INTRODUCTION: The use of peppermint oil in treating the irritable bowel syndrome has been studied with variable results probably due to the presence of patients affected by small intestinal bacterial overgrowth, lactose intolerance or celiac disease that may have symptoms similar to irritable bowel syndrome. AIM: The aim of the study was to test the effectiveness of enteric-coated peppermint oil in patients with irritable bowel syndrome in whom small intestinal bacterial overgrowth, lactose intolerance and celiac disease were excluded. METHODS: Fifty seven patients with irritable bowel syndrome according to the Rome II criteria, with normal lactose and lactulose breath tests and negative antibody screening for celiac disease, were treated with peppermint oil (two enteric-coated capsules twice per day or placebo) for 4 weeks in a double blind study. The symptoms were assessed before therapy (T(0)), after the first 4 weeks of therapy (T(4)) and 4 weeks after the end of therapy (T(8)). The symptoms evaluated were: abdominal bloating, abdominal pain or discomfort, diarrhoea, constipation, feeling of incomplete evacuation, pain at defecation, passage of gas or mucus and urgency at defecation. For each symptom intensity and frequency from 0 to 4 were scored. The total irritable bowel syndrome symptoms score was also calculated as the mean value of the sum of the average of the intensity and frequency scores of each symptom. RESULTS: At T(4), 75% of the patients in the peppermint oil group showed a >50% reduction of basal (T(0)) total irritable bowel syndrome symptoms score compared with 38% in the placebo group (P<0.009). With peppermint oil at T(4) and at T(8) compared with T(0) a statistically significant reduction of the total irritable bowel syndrome symptoms score was found (T(0): 2.19+/-0.13, T(4): 1.07+/-0.10*, T(8): 1.60+/-0.10*, *P<0.01 compared with T(0), mean+/-S.E.M.), while no change was found with the placebo. CONCLUSION: A 4 weeks treatment with peppermint oil improves abdominal symptoms in patients with irritable bowel syndrome. PMID- 17420160 TI - Endoscopic balloon dilatation applied for the treatment of ileocecal valve stricture caused by tuberculosis. PMID- 17420161 TI - The attachment of catalase and poly-l-lysine to plasma immersion ion implantation treated polyethylene. AB - Plasma immersion ion implantation (PIII) treatment of polyethylene increased the functional attachment of catalase and increased the retention of enzyme activity in comparison to untreated controls. The attached protein was not removed by SDS or NaOH, while that on the untreated surfaces was easily removed. Poly-l-lysine was found to attach in a similar way to the treated surface and could not be removed by NaOH, while it did not attach to the untreated surface. This indicates that a new binding mechanism, covalent in nature, is introduced by the plasma treatment. Surfaces treated with PIII maintained the catalase activity more effectively than surfaces plasma treated without PIII. The PIII-treated surface was hydrophilic compared to the untreated surface and retained its hydrophilic character better than surfaces subjected to a conventional plasma treatment process. The strong modification of a deeper region of the polymer than for conventional plasma treatments is believed to be responsible for both the enhanced hydrophilic character and for the increase in functional lifetime of the attached protein. The results show that PIII treatment of polymers increases their usefulness for protein microarrays. PMID- 17420162 TI - Fracture resistance of short, randomly oriented, glass fiber-reinforced composite premolar crowns. AB - The aim of this work was to determine the static load-bearing capacity of posterior composite crowns made of experimental composite resin (FC) with short fiber fillers and a semi-interpenetrating polymer network (IPN) matrix. In addition, we wanted to investigate how load-bearing capacity of surface composite resins was affected by substructures of fiber-reinforced composite (FRC) and FC, and by different curing systems. Five groups of crowns were fabricated (n=6). The crowns were either polymerized with a hand-light curing unit (LCU) or cured in a vacuum curing device (VLC) before they were statically loaded at a speed of 1mm min(-1) until fracture. Failure modes were visually examined. Data were analyzed using ANOVA. ANOVA revealed that crowns made from the FC had a statistically significant higher load-bearing capacity than the control PFC composite. Crowns with FRC substructure and PFC covering gave force values of 348N (LCU) and 1199N (VLC), respectively, which were lower than the values of FC composite. No statistically significant difference was found between crowns made from plain FC composite and those made from FC composite with a surface layer of PFC (P=0.892 and 1.00). Restorations made from short glass fiber-containing composite resin with IPN-polymer matrix showed better load bearing capacity than those made with either plain PFC or PFC reinforced with FRC substructure. PMID- 17420163 TI - Gene expression fingerprinting for human hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) or Osler-Weber-Rendu syndrome is an autosomal dominant vascular disorder characterized by telangiectases and internal arteriovenous malformations. It is caused by mutations in elements of the transforming growth factor-beta (TGF-beta) receptor complex: endoglin, a co receptor, responsible for HHT1, or ALK1 (activin receptor-like kinase 1), a type I receptor leading to HHT2. Recently, we have established cultures of HHT endothelial cells, primary targets of the disease. These cells showed deficient TGF-beta signaling and angiogenesis, representing a useful human model to study the molecular mechanism of this disease. To understand the pathogenic mechanism underlying HHT, we have used total RNA probes to compare HHT versus non-HHT cells by expression microarrays. This work represents a systematic study to identify target genes affected in HHT cells. Given the similarity of symptoms in HHT1 and HHT2, special interest has been put on the identification of common targets for both HHT types. As a result, 277 downregulated and 63 upregulated genes were identified in HHT versus control cells. These genes are involved in biological processes relevant to the HHT pathology, such as angiogenesis, cytoskeleton, cell migration, proliferation and NO synthesis. The type of misregulated genes found in HHT endothelial cells lead us to propose a model of HHT pathogenesis, opening new perspectives to understand this disorder. Moreover, as the disease is originated by mutations in proteins of the TGF-beta receptor complex, these results may be useful to find out targets of the TGF-beta pathway in endothelium. PMID- 17420164 TI - Time trends of mortality from Crohn's disease and ulcerative colitis. AB - AIMS: The present study served to test whether Crohn's disease and ulcerative colitis showed similarities in the temporal variation of their mortality rates among different countries. METHODS: Mortality data from 21 different countries between 1951 and 2005 were analysed, including Argentina, Australia, Austria, Belgium, Canada, Chile, Denmark, England, Finland, France, Germany, Italy, Japan, Mexico, Netherlands, Scotland, Spain, Sweden, Switzerland, Taiwan and USA. The age-specific death rates of each individual country, as well as the average age specific rates of all countries, were plotted against the period of death. RESULTS: Death rates from ulcerative colitis were initially 6-fold higher than those of Crohn's disease. Mortality from ulcerative colitis decreased continuously during the past 50 years. Mortality from Crohn's disease increased from 1951 to 1975 until reaching a similar level as mortality from ulcerative colitis. Since then the death rates of both diseases have followed a parallel time course. A same type of behaviour was found in the time trends of each individual age-group. The data from most countries revealed similar temporal patterns. CONCLUSIONS: The relationships between the temporal changes of mortality from Crohn's disease and ulcerative colitis might suggest the presence of one primary risk factor responsible for the occurrence of both diseases, and at the existence of one additional secondary risk factor, responsible for the expression of Crohn's disease alone. PMID- 17420165 TI - Comparison of two survey methodologies to assess vaccination coverage. AB - BACKGROUND: Measuring vaccination coverage permits evaluation and appropriate targeting of vaccination services. The cluster survey methodology developed by the World Health Organization, known as the 'Expanded Program on Immunization (EPI) methodology', has been used worldwide to assess vaccination coverage; however, the manner in which households are selected has been criticized by survey statisticians as lacking methodological rigor and introducing bias. METHODS: Thirty clusters were selected from an urban (Ambo) and a rural (Yaya Gulelena D/Libanos) district of Ethiopia; vaccination coverage surveys were conducted using both EPI sampling and systematic random sampling (SystRS) of households. Chi-square tests were used to compare results from the two methodologies; relative feasibility of the sampling methodologies was assessed. RESULTS: Vaccination coverage from a recent measles campaign among children aged 6 months through 14 years was high: 95% in Ambo (both methodologies), 91 and 94% (SystRS and EPI sampling, respectively, P-value = 0.05) in Yaya-Gulelena D/Libanos. Coverage with routine vaccinations among children aged 12-23 months was <20% in both districts; in Ambo, EPI sampling produced consistently higher estimates of routine coverage than SystRS. Differences between the two methods were found in demographic characteristics and recent health histories. Average time required to complete a cluster was 16h for EPI sampling and 17 h for SystRS; total cost was equivalent. Interviewers reported slightly more difficulty conducting SystRS. CONCLUSIONS: Because of the methodological advantages and demonstrated feasibility, SystRS would be preferred to EPI sampling in most situations. Validating results in additional settings is recommended. PMID- 17420166 TI - Anaemia of CKD--the CHOIR study revisited. PMID- 17420167 TI - Prevalence and clinical correlates of white coat hypertension in chronic kidney disease. AB - BACKGROUND: The role of white coat hypertension (WCH) in the poor control of blood pressure (BP) in chronic kidney disease (CKD) is ill defined. METHODS: We measured systolic clinical (CBP) and ambulatory blood pressure (ABP) in 290 consecutive patients with non-dialysis CKD [glomerular filtration rate (GFR) <60 ml/min/1.73 m(2)]. We defined normotension (NOR) if CBP and daytime ABP <130 mmHg, sustained hypertension (SH) when both BP >or=130 mmHg, WCH if only daytime ABP <130 mmHg, and masked hypertension (MH) when only CBP <130 mmHg. RESULTS: NOR patients were 15.5%, WCH 31.7%, SH 46.9% and MH 5.9%. Due to the high prevalence of WCH, achievement of BP target (<130 mmHg) was more than doubled by daytime ABP than CBP (47.2 vs 21.4%). WCH was characterized by prevalence of diabetes (31.5%), left ventricular hypertrophy (LVH; 50.0%) and CBP values (146 +/- 12 mmHg) lower than in SH (41.9%, 71.3% and 158 +/- 18 mmHg) but greater than in NOR (17.8%, 37.8% and 118 +/- 7 mmHg). Among patients with CBP >or=130 mmHg, the independent risk of having SH rather than WCH increased in the presence of higher CBP [Odds ration (OR) 1.61, 95% confidence intervals (CI) 1.29-2.02], LVH (OR 1.94, 95% CI 1.03-3.63) and proteinuria (OR 3.12, 95% CI 1.31-7.43). In the WCH group, 24 h, daytime and nighttime ABP were 118 +/- 7/68 +/- 8, 120 +/- 7/71 +/- 8 and 112 +/- 12/63 +/- 9 mmHg, respectively. CONCLUSIONS: In CKD, WCH is highly prevalent and can be predicted in the absence of higher CBP, LVH and proteinuria. In these patients, pursuing a low BP target may not be safe because of the risk of cardio-renal hypoperfusion especially at nighttime. PMID- 17420168 TI - Reduced systemic advanced glycation end products in children receiving peritoneal dialysis with low glucose degradation product content. AB - BACKGROUND: Glucose degradation products (GDP) in peritoneal dialysis (PD) solutions are toxic to the peritoneal membrane and promote the formation of advanced glycation end products (AGE), which contribute to accelerated atherosclerosis and amyloidosis. Double chamber PD solutions have a markedly reduced GDP content. METHODS: We analysed GDP and AGE kinetics in 21 children (7 months to 18 years) on automated PD in a prospective multicentre trial with randomized administration of single chamber, high-GDP and double-chamber, low-GDP dialysis solution for 12 weeks each. Total AGE fluorescence, carboxymethyllysine (CML, ELISA) and 3-deoxyglucosone (3-DG, HPLC) were measured in plasma and PD effluent during a 4 h peritoneal equilibration test. Plasma AGE profiles were assessed by size selective gel permeation chromatography and compared with 23 healthy controls. RESULTS: Initial effluent 3-DG concentrations were 140+/-55 and 25+/-4 micromol/l with high- and low-GDP PD fluid, respectively and declined to 53+/-32 and 7+/-2 micromol/l within 4 h dwell time (P<0.001). The ex vivo AGE generating capacity was three times higher with the high-GDP solution and decreased significantly with dwell time. Plasma AGE levels were 1.8-7.4-fold above those of healthy controls; the elevation was most marked for the small molecular fraction (<2 kDa). Plasma AGE and CML levels were significantly higher after 12 weeks exposure to high-GDP solution (20991+/-4145 AU and 1505+/-617 ng/ml) than following treatment with low-GDP fluid (17518+/-4676 AU and 1151+/ 438 ng/ml; both P<0.05). Four hour AGE clearance was higher with low-GDP solution (0.74+/-0.3 vs 0.44+/-0.15 ml/min*1.73 m2, P<0.01). CONCLUSION: GDP are rapidly absorbed from the peritoneal cavity. Administration of PD solutions with low-GDP content reduces plasma AGE levels and may thus improve the cardiovascular risk profile of dialysed children. PMID- 17420169 TI - Apo-10'-lycopenoic acid inhibits lung cancer cell growth in vitro, and suppresses lung tumorigenesis in the A/J mouse model in vivo. AB - High intake of lycopene has been associated with a lower risk of a variety of cancers including lung cancer. We recently showed that lycopene can be converted to apo-10'-lycopenoids [Hu et al. (2006). J. Biol. Chem., 281, 19327-19338] in mammalian tissues both in vitro and in vivo, raising the question of whether apo 10'-lycopenoids have biological activities against lung carcinogenesis. In the present study, we report that apo-10'-lycopenoic acid inhibited the growth of NHBE normal human bronchial epithelial cells, BEAS-2B-immortalized normal bronchial epithelial cells and A549 non-small cell lung cancer cells. This inhibitory effect of apo-10'-lycopenoic acid was associated with decreased cyclin E, inhibition of cell cycle progression from G(1) to S phase and increased cell cycle regulators p21 and p27 protein levels. In addition, apo-10'-lycopenoic acid transactivated the retinoic acid receptor beta (RARbeta) promoter and induced the expression of RARbeta. We further examined the effect of apo-10'-lycopenoic acid treatment on 4-(N-methyl-N-nitrosamino)-1-(3-pyridal)-1-butanone (NNK)-induced lung tumorigenesis in the A/J mouse model. We found that the lung tumor multiplicity was decreased dose dependently from an average of 16 tumors per mouse in the NNK injection alone group, to an average of 10, 7 and 5 tumors per mouse in groups injected with NNK and supplemented with 10, 40 and 120 mg/kg diet of apo-10'-lycopenoic acid, respectively. These observations demonstrate that apo 10'-lycopenoic acid is a biological active metabolite of lycopene and suggest that apo-10'-lycopenoic acid is a potential chemopreventive agent against lung tumorigenesis. PMID- 17420170 TI - The functional genetic variant Arg324Gly of frizzled-related protein is associated with colorectal cancer risk. AB - The Wnt-beta-catenin pathway plays a central role in colorectal tumorigenesis. Frizzled-related protein (FRZB, also termed secreted frizzled-related protein 3, sFRP3) antagonizes the signaling of wingless (Wnt) ligands through the frizzled membrane-bound receptors, resulting in beta-catenin destabilization thereby suppressing the expression of target genes. Recently, the FRZB Gly324 variant has been shown to have an attenuated ability to antagonize Wnt signaling and to be associated with an increased osteoarthritis risk. Here, we investigated, for the first time, the role of Arg324Gly (970C>G) along with Arg200Trp (598C>T) on colorectal cancer (CRC) risk by analyzing 659 patients and 607 control individuals drawn from the German DACHS (Darmkrebs: Chancen der Verhutung durch Screening) study. Although Arg200Trp showed no effect on CRC risk, we found homozygous carriers of Gly324 more frequent in cases than in controls, leading to a significantly increased risk for CRC [odds ratio (OR) = 5.1, 95% confidence interval (95% CI) = 1.74-14.71, P < 0.001]. The association was stronger in rectal cancer (OR = 7.52, 95% CI = 2.40-23.25, P < 0.0001) than in colon cancer (OR = 3.66, 95% CI = 1.14-11.76, P < 0.05). Since modified Wnt signaling and down regulation of frizzled-related proteins have been observed in many human cancers, this variant may also affect the susceptibility to other cancers. PMID- 17420171 TI - Stress-driven in vivo selection of a functional mini-gene from a randomized DNA library expressing combinatorial peptides in Escherichia coli. AB - A plasmid-borne randomized mini-gene library expressing a population of combinatorial 20-mer peptides with no bias toward any biological function was used as an initial source of genetic variance during stress-driven evolution of Escherichia coli. The transformed bacteria were evolved under multiple rounds of selective pressure imposed by nearly lethal concentrations of NiCl(2), AgNO(3), or K(2)TeO(3). At the final stage, clones conferring resistance to NiCl(2) were found to carry identical functional mini-genes, which conferred significant nickel tolerance on the host cells. Expression of the mini-gene markedly improved growth parameters of the evolved clones at subinhibitory concentrations of NiCl(2) while being slightly detrimental in the absence of the stress. This substantial increase in resistance, as compared with control cultures adapted in the absence of the mini-gene, is shown to be largely due to coadaptation with changes elsewhere in the E. coli genome. Clones resistant to AgNO(3) and K(2)TeO(3) harbored plasmid variants with an inactive mini-gene and with a deleted mini-gene operon, respectively. In those cases, an exploration of the mini-gene sequence space apparently was fruitless, and the developed toxicity tolerance was likely to be exclusively associated with acquired adaptive mutations. Overall, the results demonstrate a very natural outcome in which the mini-genes were expected to be either successfully integrated into the bacterial genetic network or rejected depending on their effect on host fitness. This approach is immediately useful as a laboratory model to study the dynamics of bacterial adaptive evolution at the molecular level and is especially relevant as a rapid method to study cellular response to recently acquired genetic material. PMID- 17420172 TI - Local potential connectivity in cat primary visual cortex. AB - Time invariant description of synaptic connectivity in cortical circuits may be precluded by the ongoing growth and retraction of dendritic spines accompanied by the formation and elimination of synapses. On the other hand, the spatial arrangement of axonal and dendritic branches appears stable. This suggests that an invariant description of connectivity can be cast in terms of potential synapses, which are locations in the neuropil where an axon branch of one neuron is proximal to a dendritic branch of another neuron. In this paper, we attempt to reconstruct the potential connectivity in local cortical circuits of the cat primary visual cortex (V1). Based on multiple single-neuron reconstructions of axonal and dendritic arbors in 3 dimensions, we evaluate the expected number of potential synapses and the probability of potential connectivity among excitatory (pyramidal and spiny stellate) neurons and inhibitory basket cells. The results provide a quantitative description of structural organization of local cortical circuits. For excitatory neurons from different cortical layers, we compute local domains, which contain their potentially pre- and postsynaptic excitatory partners. These domains have columnar shapes with laminar specific radii and are roughly of the size of the ocular dominance column. Therefore, connections between most excitatory neurons in the ocular dominance column can be implemented by local synaptogenesis. Structural connectivity involving inhibitory basket cells is generally weaker than excitatory connectivity. Here, only nearby neurons are capable of establishing more than one potential synapse, implying that within the ocular dominance column these connections have more limited potential for circuit remodeling. PMID- 17420173 TI - Control of flowering time in temperate cereals: genes, domestication, and sustainable productivity. AB - The control of flowering is central to reproductive success in plants, and has a major impact on grain yield in crop species. The global importance of temperate cereal crops such as wheat and barley has meant emphasis has long been placed on understanding the genetics of flowering in order to enhance yield. Leads gained from the dissection of the molecular genetics of model species have combined with comparative genetic approaches, recently resulting in the isolation of the first flowering time genes in wheat and barley. This paper reviews the genetics and genes involved in cereal flowering pathways and the current understanding of how two of the principal genes, Vrn and Ppd, have been involved in domestication and adaptation to local environments, and the implications for future breeding programmes are discussed. PMID- 17420174 TI - Root hair curling and Rhizobium infection in Medicago truncatula are mediated by phosphatidylinositide-regulated endocytosis and reactive oxygen species. AB - The symbiotic relationships between legumes and rhizobacteria involve extensive signalling between the two organisms. Studies using genetic, biochemical, and pharmacological approaches have demonstrated the involvement of calcium and reactive oxygen species in the establishment of symbiotic interactions. In the early stage of the interactions rhizobia grow as infection thread within host root hairs and are internalized into the plant cells via endocytosis. It is shown here that inoculation of Medicago truncatula roots with Sinorhizobium meliloti induced a battery of vesicle trafficking genes, including the phosphatidylinositol 3-kinase (PI3K) gene that stimulated plasma membrane endocytosis and the production of reactive oxygen species (ROS). Inhibition of the PI3K suppressed the membrane endocytosis and subsequent oxidative burst and prevented root hair curling and formation of infection threads. Similar effects were produced by inhibition of PtdIns-specific phospholipase C (PI-PLC). However, neither inhibition of PI3K nor PI-PLC signalling blocked cytosolic Ca2+ influx or early nodulin (ENOD) gene expression. By contrast, the inhibitors induced ENODs transcription in the absence of Rhizobium, suggesting that the expression of ENODs responds to plasma membrane perturbations. In summary, the results show a major reprogramming of intracellular vesicle trafficking during the early stages of symbiotic interactions that co-ordinate the host responses. Activation of parallel signalling pathways leading to Cacyt2+ influx and ROS production that regulate the root hair curling and ENODs expression are also shown. PMID- 17420175 TI - Clinicians' concepts of the cognitive deficits of schizophrenia. AB - Several compounds to improve cognition in schizophrenia are being studied in clinical trials, but little is known about how clinicians conceptualize the cognitive deficits of schizophrenia. In a pilot study, the author asked 40 psychiatrists 3 brief questions about the clinical presentation of cognitive deficits. Descriptions of cognitive deficits show high variability. Informants describe phenomenology like follow-through, attention, and emptiness as indicative of cognitive impairment. Informants' concepts of cognitive deficits overlap substantially with positive, negative, and thought disorder symptoms. Clinicians' concepts are complex and contextualized, in contrast to the discrete skills measured by neuropsychological tests. Results suggest that appropriate prescribing of cognition-enhancing medications may be challenging. PMID- 17420176 TI - Beating the odds--nothing is impossible, its just a road less traveled. AB - By contrasting students with learning disabilities and students with schizophrenia, it becomes conspicuously clear that they face many of the same hurdles. I know through personal experience because I have both. Postsecondary institutions are making great strides in balancing the scales for disabled students. Special education for learning-disabled students is more a hindrance than a benefit; mainstreaming many learning-disabled students seems a more appropriate option. Students need to take advantage of the services that are most beneficial to them. The least restrictive environment is predominantly, if not always, the correct choice. PMID- 17420177 TI - The course of neurocognition and social functioning in individuals at ultra high risk for psychosis. AB - OBJECTIVE: This study evaluates longitudinal neuropsychological performance and its association with clinical symptomatology and psychosocial outcome in individuals identified as ultra high risk (UHR) for psychosis. METHODS: Thirty five UHR individuals completed neurocognitive, clinical, and social/role functioning assessments at baseline and, on average, 8.3 months later. RESULTS: UHR subjects showed significant cognitive deficits at baseline and 2 distinct profiles of cognitive change over time. On average, 50% demonstrated improvement in social and role functioning over the follow-up period, while the other half showed either stability or decline in functioning. Functional improvement was associated with improved processing speed and visual memory, as well as improvement in clinical symptoms over the follow-up period. In contrast, patients who did not improve functionally showed stable clinical symptoms and cognitive performance over time. CONCLUSIONS: Although the degree of neurocognitive deficit at baseline in UHR patients does not predict psychosocial outcome, the course of neurocognitive change over the first 8 months of follow-up does differentiate patients with good and poor functional outcomes. PMID- 17420178 TI - Microsatellite isolation and linkage group identification in the yellow fever mosquito Aedes aegypti. AB - Microsatellites have proved to be very useful as genetic markers, as they seem to be ubiquitous and randomly distributed throughout most eukaryote genomes. However, our laboratories and others have determined that this paradigm does not necessarily apply to the yellow fever mosquito Aedes aegypti. We report the isolation and identification of microsatellite sequences from multiple genomic libraries for A. aegypti. We identified 6 single-copy simple microsatellites from 3 plasmid libraries enriched for (GA)(n), (AAT)(n), and (TAGA)(n) motifs from A. aegypti. In addition, we identified 5 single-copy microsatellites from an A. aegypti cosmid library. Genetic map positions were determined for 8 microsatellite loci. These markers greatly increase the number of microsatellite markers available for A. aegypti and provide additional tools for studying genetic variability of mosquito populations. Additionally, most A. aegypti microsatellites are closely associated with repetitive elements that likely accounts for the limited success in developing an extensive panel of microsatellite marker loci. PMID- 17420179 TI - Analysis of flavonoids in flower petals of soybean near-isogenic lines for flower and pubescence color genes. AB - W1, W3, W4, and Wm genes control flower color, whereas T and Td genes control pubescence color in soybean. W1, W3, Wm, and T are presumed to encode flavonoid 3'5'-hydroxylase (EC 1.14.13.88), dihydroflavonol 4-reductase (EC 1.1.1.219), flavonol synthase (EC 1.14.11.23), and flavonoid 3'-hydroxylase (EC 1.14.13.21), respectively. The objective of this study was to determine the structure of the primary anthocyanin, flavonol, and dihydroflavonol in flower petals. Primary component of anthocyanin in purple flower cultivars Clark (W1W1 w3w3 W4W4 WmWm TT TdTd) and Harosoy (W1W1 w3w3 W4W4 WmWm tt TdTd) was malvidin 3,5-di-O-glucoside with delphinidin 3,5-di-O-glucoside as a minor compound. Primary flavonol and dihydroflavonol were kaempferol 3-O-gentiobioside and aromadendrin 3-O-glucoside, respectively. Quantitative analysis of near-isogenic lines (NILs) for flower or pubescence color genes, Clark-w1 (white flower), Clark-w4 (near-white flower), Clark-W3w4 (dilute purple flower), Clark-t (gray pubescence), Clark-td (near-gray pubescence), Harosoy-wm (magenta flower), and Harosoy-T (tawny pubescence) was carried out. No anthocyanins were detected in Clark-w1 and Clark-w4, whereas a trace amount was detected in Clark-W3w4. Amount of flavonols and dihydroflavonol in NILs with w1 or w4 were largely similar to the NILs with purple flower suggesting that W1 and W4 affect only anthocyanin biosynthesis. Amount of flavonol glycosides was substantially reduced and dihydroflavonol was increased in Harosoy-wm suggesting that Wm is responsible for the production of flavonol from dihydroflavonol. The recessive wm allele reduces flavonol amount and inhibits co-pigmentation between anthocyanins and flavonols resulting in less bluer (magenta) flower color. Pubescence color genes, T or Td, had no apparent effect on flavonoid biosynthesis in flower petals. PMID- 17420180 TI - Aging, retirement, and changes in physical activity: prospective cohort findings from the GLOBE study. AB - There is increased recognition that determinants of health should be investigated in a life-course perspective. Retirement is a major transition in the life course and offers opportunities for changes in physical activity that may improve health in the aging population. The authors examined the effect of retirement on changes in physical activity in the GLOBE Study, a prospective cohort study known by the Dutch acronym for "Health and Living Conditions of the Population of Eindhoven and surroundings," 1991-2004. They followed respondents (n = 971) by postal questionnaire who were employed and aged 40-65 years in 1991 for 13 years, after which they were still employed (n = 287) or had retired (n = 684). Physical activity included 1) work-related transportation, 2) sports participation, and 3) nonsports leisure-time physical activity. Multinomial logistic regression analyses indicated that retirement was associated with a significantly higher odds for a decline in physical activity from work-related transportation (odds ratio (OR) = 3.03, 95% confidence interval (CI): 1.97, 4.65), adjusted for sex, age, marital status, chronic diseases, and education, compared with remaining employed. Retirement was not associated with an increase in sports participation (OR = 1.12, 95% CI: 0.71, 1.75) or nonsports leisure-time physical activity (OR = 0.80, 95% CI: 0.54, 1.19). In conclusion, retirement introduces a reduction in physical activity from work-related transportation that is not compensated for by an increase in sports participation or an increase in nonsports leisure-time physical activity. PMID- 17420181 TI - A prospective study of tobacco, alcohol, and the risk of esophageal and gastric cancer subtypes. AB - Rates of esophageal adenocarcinoma and gastric cardia adenocarcinoma have increased, while rates of esophageal squamous cell carcinoma (ESCC) and gastric noncardia adenocarcinoma have decreased, suggesting distinct etiologies. The authors prospectively investigated the associations of alcohol and tobacco with these cancers in 474,606 US participants using Cox models adjusted for potential confounders. Between 1995/1996 and 2000, 97 incident cases of ESCC, 205 of esophageal adenocarcinoma, 188 of gastric cardia, and 187 of gastric noncardia cancer occurred. Compared with nonsmokers, current smokers were at increased risk for ESCC (hazard ratio (HR) = 9.27, 95% confidence interval (CI): 4.04, 21.29), esophageal adenocarcinoma (HR = 3.70, 95% CI: 2.20, 6.22), gastric cardia (HR = 2.86, 95% CI: 1.73, 4.70), and gastric noncardia (HR = 2.04, 95% CI: 1.32, 3.16). Assuming causality, ever smoking had population attributable risks of 77% (95% CI: 0.55, 0.89) for ESCC, 58% (95% CI: 0.38, 0.72) for esophageal adenocarcinoma, 47% (95% CI: 0.27, 0.63) for gastric cardia, and 19% (95% CI: 0.00, 0.37) for gastric noncardia. For drinkers of more than three alcoholic beverages per day, compared with those whose intake was up to one drink per day, the authors found significant associations between alcohol intake and ESCC risk (HR = 4.93, 95% CI: 2.69, 9.03) but not risk for esophageal, gastric cardia, or gastric noncardia adenocarcinoma. PMID- 17420182 TI - Efficacy of the male latex condom and of the female polyurethane condom as barriers to semen during intercourse: a randomized clinical trial. AB - In this 2000-2001 study, the authors compared the effectiveness of the male latex condom and the female polyurethane condom by assessing frequency and types of mechanical failure and by evaluating semen exposure during use. Eligible women from Birmingham, Alabama, were randomly assigned to begin the study with 10 male condoms and then switch to 10 female condoms (n = 55), or vice versa (n = 53), and were trained to use both types. Data collection included questionnaires for each condom use and measurement of prostate-specific antigen in specimens of vaginal fluid taken before and after intercourse. Participants returned 700 male condoms and 678 female condoms, and they reported mechanical problems for 9% and 34%, respectively. Moderate-high postcoital prostate-specific antigen levels (> or = 22 ng/ml) were detected in 3.5% of male condom uses and 4.5% of female condom uses (difference = 1%, 95% confidence interval: -1.6, 3.7). Moderate-high prostate-specific antigen values (> or = 22 ng/ml) were more frequent with mechanical problems (male condom, 9.6%; female condom, 9.4%) but less frequent with other problems (3.0% and 0.9%) or correct use with no problems (2.7% and 2.5%). This study indicates that although mechanical problems are more common with the female condom than with the male condom, these devices may involve a similar risk of semen exposure. Objectively assessed semen exposure is associated with self-reported mechanical problems. PMID- 17420183 TI - CpG methylation is targeted to transcription units in an invertebrate genome. AB - DNA is methylated at the dinucleotide CpG in genomes of a wide range of plants and animals. Among animals, variable patterns of genomic CpG methylation have been described, ranging from undetectable levels (e.g., in Caenorhabditis elegans) to high levels of global methylation in the vertebrates. The most frequent pattern in invertebrate animals, however, is mosaic methylation, comprising domains of methylated DNA interspersed with unmethylated domains. To understand the origin of mosaic DNA methylation patterns, we examined the distribution of DNA methylation in the Ciona intestinalis genome. Bisulfite sequencing and computational analysis revealed methylated domains with sharp boundaries that strongly colocalize with approximately 60% of transcription units. By contrast, promoters, intergenic DNA, and transposons are not preferentially targeted by DNA methylation. Methylated transcription units include evolutionarily conserved genes, whereas the most highly expressed genes preferentially belong to the unmethylated fraction. The results lend support to the hypothesis that CpG methylation functions to suppress spurious transcriptional initiation within infrequently transcribed genes. PMID- 17420184 TI - The mode and tempo of genome size evolution in eukaryotes. AB - Eukaryotic genome size varies over five orders of magnitude; however, the distribution is strongly skewed toward small values. Genome size is highly correlated to a number of phenotypic traits, suggesting that the relative lack of large genomes in eukaryotes is due to selective removal. Using phylogenetic contrasts, we show that the rate of genome size evolution is proportional to genome size, with the fastest rates occurring in the largest genomes. This trend is evident across the 20 major eukaryotic clades analyzed, indicating that over long time scales, proportional change is the dominant and universal mode of genome-size evolution in eukaryotes. Our results reveal that the evolution of eukaryotic genome size can be described by a simple proportional model of evolution. This model explains the skewed distribution of eukaryotic genome sizes without invoking strong selection against large genomes. PMID- 17420185 TI - Scientists examine pain relief and addiction. PMID- 17420187 TI - Prevention of respiratory depression becomes safety foundation's new goal. PMID- 17420189 TI - FDA's sentinel network: a monumental task. PMID- 17420190 TI - FDA investigating nonprescription cough and cold products. PMID- 17420191 TI - Effect of tobacco on INR. PMID- 17420192 TI - ASHP therapeutic position statement on the use of second-generation antipsychotic medications in the treatment of adults with psychotic disorders. PMID- 17420193 TI - Clinical pharmacy services in a home-based palliative care program. PMID- 17420194 TI - Specializing in pediatric hematology and oncology. PMID- 17420195 TI - The role of second-generation antipsychotics in the treatment of psychotic disorders. PMID- 17420196 TI - Ensuring the competence of pharmacy technicians. PMID- 17420197 TI - Polymyxin antibiotics for gram-negative infections. AB - PURPOSE: The role of polymyxin antibiotics in the treatment of multidrug resistant gram-negative infections is reviewed. SUMMARY: Antimicrobial resistance is an increasing problem across hospitals worldwide, especially in intensive care settings, where nosocomial infections are 5-10 times more likely to occur than on the general wards. The polymyxins, a group of basic polypeptide antibiotics, were first isolated from Bacillus species in the late 1940s and appear to have a surface detergent effect, making them active against most gram-negative organisms. Early clinical reports suggested a high rate of toxicity associated with the polymyxins, specifically nephrotoxicity (20%) and neurotoxicity (7%); thus the polymyxins had largely fallen out of favor. However, recent studies have suggested that the toxicities associated with the polymyxins may be less severe and less frequent than earlier reports. The emergence of multidrug-resistant gram negative organisms has led to a reemergence in the use of this antibiotic class. Various clinical trials that evaluated the polymyxins for the treatment of multidrug-resistant gram-negative organisms found that these antibiotics have acceptable effectiveness and may be used if necessary. CONCLUSION: The polymyxins have become a last resort for the treatment of infections caused by multidrug resistant gram-negative organisms. Recent studies have suggested that the frequency of polymyxin-associated nephrotoxicity and neurotoxicity may not be as high as was once thought. The polymyxins seem to be effective in treating various infections caused by multidrug-resistant gram-negative organisms but should not be used as first-line therapy until more is known about this class of antibiotics. PMID- 17420198 TI - Potential aripiprazole-mediated extrapyramidal symptoms in an adult with developmental disabilities. AB - PURPOSE: A case of extrapyramidal symptoms (EPS) following administration of aripiprazole to a man with developmental disabilities who had never received antipsychotic medications and had no history of movement disorders is presented. SUMMARY: The patient was a 40-year-old male with developmental disabilities. He was nonverbal, profoundly mentally retarded, and diagnosed with obsessive compulsive disorder (OCD) and orthopedic problems. He developed episodic movements possibly consistent with EPS secondary to aripiprazole usage. The patient was antipsychotic naive before initiation of aripiprazole 5 mg daily. Concurrent medications at the time of EPS onset included oxazepam, baclofen, and citalopram. Baclofen and oxazepam were prescribed secondary to right-sided hemiparesis contractures. Aripiprazole, 5 mg daily, was initiated in November 2004 as an augmentation strategy for the diagnosis of OCD. Facial, tongue, and arm movements were first reported approximately five weeks after the initiation of aripiprazole. Initial symptoms resolved after approximately 24 hours. The dosage was increased to 10 mg daily two weeks later. Dystonic episodes continued on an intermittent basis, and the patient presented with lower-lip thrusting and upper-limb athetosis. These movements interfered with the patient's eating, chewing, and holding of utensils. Several of the standard treatment strategies for EPS were used. Initially, diphenhydramine hydrochloride 25 mg was administered orally every six hours. The patient's movements resolved following diphenhydramine administration. Aripiprazole was subsequently discontinued secondary to its lack of efficacy for OCD and the development of a movement disorder. CONCLUSION: A patient with developmental disabilities who had no history of movement disorders developed EPS following initiation of aripiprazole. PMID- 17420199 TI - Azithromycin for the secondary prevention of coronary artery disease: a meta analysis. AB - PURPOSE: A meta-analysis of randomized, controlled trials that evaluated the effect of the macrolide antibiotic, azithromycin, on clinical outcomes in patients with coronary artery disease (CAD) was conducted. METHODS: A systematic literature search of MEDLINE, EMBASE, Web of Science, and the Cochrane Database of Systematic Reviews was conducted using specific search terms. Randomized, controlled trials comparing azithromycin or placebo in secondary CAD patients with adequately reported data on mortality and clinical cardiac endpoints were included. A random-effects model was used. RESULTS: Six studies (n=13,778) met the inclusion criteria. The trials varied in their design. On meta-analysis, azithromycin resulted in a nonsignificant reduction in mortality versus placebo (odds ratio [OR], 0.91; 95% confidence interval [CI], 0.77-1.09; p=0.31). Four trials reported the rate of nonfatal myocardial infarction (MI). Azithromycin did not have an effect on the rate of nonfatal MI versus placebo (OR, 0.95; 95% CI, 0.80-1.13; p=0.57). Five trials reported rates of hospitalization in which no significant difference was seen with azithromycin versus placebo (OR, 0.97; 95% CI, 0.80-1.17; p=0.76). Six trials were used to evaluate the composite cardiovascular endpoint. Again, no significant benefit was seen with azithromycin versus placebo (OR, 0.93; 95% CI, 0.84-1.03; p=0.218). CONCLUSION: Meta-analysis showed that azithromycin does not appear to reduce the frequency of recurrent cardiac events in patients with CAD. Results from ongoing trials may clarify the role of azithromycin in the secondary prevention of coronary events. PMID- 17420200 TI - Effect of two work practice changes on the microbial contamination rates of pharmacy-compounded sterile preparations. AB - PURPOSE: Using a multiple-step testing medium-risk-level compounding test procedure, the evaluation of two work-practice changes to determine if the changes could effectively reduce the potential for contamination occurrence was conducted. SUMMARY: Along with training and evaluation of aseptic sterile compounding techniques, each individual pharmacist and pharmacy technician at M. D. Anderson Cancer Center must successfully demonstrate aseptic preparation competency annually by performing the complicated multistep aseptic transfers of growth medium with no resulting growth of microorganisms. The multistep aseptic transfers are designed to simulate manual compounding of the most complicated medium-risk-level preparations anticipated as specified in the United States Pharmacopeia's chapter 797. An evaluation of two modest and simple work-practice changes was conducted: The use of bare hands and nonsterile gloves with only initial disinfection with 70% isopropyl alcohol (IPA) during years 1 and 2 (group A) was compared with the use of nonsterile chemotherapy gloves with initial and repeated disinfection with IPA for year 3 (group B) and the use of sterile gloves with initial and repeated disinfection with IPA for year 4 (group C). The process involved multiple discrete manipulations, including reconstitution of dry-growth medium; transfers of growth medium from vials and ampules using syringes, needles, a dispensing pin, and a filter straw; and transfers to an empty plastic i.v. bag. For groups B and C, significant reductions in contaminated samples were found compared with group A. CONCLUSION: The use of protective chemotherapy gloves that were repeatedly disinfected with IPA decreased the contamination rate of pharmacy-compounded sterile preparations. PMID- 17420201 TI - Pharmacists versus nonpharmacists in adverse drug event detection: a meta analysis and systematic review. AB - PURPOSE: A systematic review and metaanalysis were conducted to determine if studies that included pharmacists as chart reviewers detected higher rates of adverse drug events (ADEs) than studies that included other health care professionals or hospital personnel as chart reviewers. METHODS: A systematic review and metaanalysis of studies using chart review as the method of detection of ADEs were conducted. Pooled estimates of the ADE rates were calculated using the inverse variance weight method. Meta-analysis was performed using a random effects model. Using the Mann-Whitney U test, weighted rates of studies in which pharmacists versus other clinicians were the chart reviewers were compared. RESULTS: Thirteen studies satisfied the inclusion criteria. Using random effects metaanalysis, the mean of the weighted incidence rate detected by pharmacists was 0.33 ADE per admission (95% confidence interval [CI], 0.17-0.50); the mean was 0.16 ADE per admission (95% CI, 0.11-0.22) with detection by nonpharmacists. Significant heterogeneity was present between studies in both groups. A significant difference (p=0.003) existed between the ADE rate reported by pharmacists (median=0.23; interquartile range [IQR], 0.18-0.44) and that of nonpharmacists (median=0.12; IQR, 0.02-0.49). Although there is overwhelming evidence of statistical heterogeneity, the numbers pertaining to the ADE rates detected by the two groups were large enough to indicate significant differences. Despite the heterogeneity, there is strong evidence that pharmacist-led interventions based on chart review report a higher ADE rate among inpatients. CONCLUSION: A review of the literature revealed that pharmacists make a salient contribution as manual chart reviewers in inpatient ADE interventions. PMID- 17420202 TI - Multidisciplinary approach to inpatient medication reconciliation in an academic setting. AB - PURPOSE: The effectiveness of a multidisciplinary medication reconciliation process was studied in an inpatient family medicine unit of an academic hospital center. METHODS: In phase 1 of this two-phase study, nurses, pharmacists, and physicians used an admission medication reconciliation form to reconcile patients' home medications on admission. The form was then reviewed by the pharmacist on the unit and by the attending physician, who reconciled the discharge medication list. The discharge medication list was compared against the patient's home medications list, inpatient medication profile, and prescriptions documented in the electronic medical record to investigate any medication discrepancies. Pharmacists participating in the study documented and categorized medication discrepancies by the potential severity of the error. In phase 2, family medicine medical residents and staff were instructed to include reconciled admission and discharge medication lists in the hospital summary. RESULTS: A total of 102 patients formed the study sample. There was no significant difference between phase 1 and phase 2 patients in mean age, sex, and length of hospital stay. Totals of 432 and 367 admission medications required reconciliation during phase 1 and phase 2, respectively. The mean number of admission medication discrepancies decreased from 0.5 per patient in phase 1 to 0 per patient in phase 2. The mean number of discharge medication discrepancies decreased from 3.3 per patient in phase 1 to 1.8 per patient in phase 2. CONCLUSION: The mean number of medication discrepancies occurring during admission and discharge decreased after a multidisciplinary medication reconciliation process was implemented in an inpatient family medicine unit of an academic hospital center. PMID- 17420203 TI - Does your pharmacy have a compounding isolator? PMID- 17420204 TI - No ordinary time. PMID- 17420205 TI - Comprehensive analysis of the effects of ordinary nutrients on hepatitis C virus RNA replication in cell culture. AB - To date, only a limited number of studies have reported finding an influence of ordinary nutrients on hepatitis C virus (HCV) RNA replication. However, the effects of other nutrients on HCV RNA replication remain largely unknown. We recently developed a reporter assay system for genome-length HCV RNA replication in hepatoma-derived HuH-7 cells (OR6). Here, using this OR6 assay system, we comprehensively examined 46 nutrients from four nutrient groups: vitamins, amino acids, fatty acids, and salts. We found that three nutrients-beta-carotene, vitamin D(2), and linoleic acid-inhibited HCV RNA replication and that their combination caused additive and/or synergistic effects on HCV RNA replication. In addition, combined treatment with each of the three nutrients and interferon alpha or beta or fluvastatin inhibited HCV RNA replication in an additive manner, while combined treatment with cyclosporine synergistically inhibited HCV RNA replication. In contrast, we found that vitamin E enhanced HCV RNA replication and negated the effects of the three anti-HCV nutrients and cyclosporine but not those of interferon or fluvastatin. These results will provide useful information for the treatment of chronic hepatitis C patients who also take anti-HCV nutrients as an adjunctive therapy in combination with interferon. In conclusion, among the ordinary nutrients tested, beta-carotene, vitamin D(2), and linoleic acid possessed anti-HCV activity in a cell culture system, and these nutrients are therefore considered to be potential candidates for enhancing the effects of interferon therapy. PMID- 17420206 TI - PhoU is a persistence switch involved in persister formation and tolerance to multiple antibiotics and stresses in Escherichia coli. AB - When a bactericidal antibiotic is added to a growing bacterial culture, the great majority of the bacterial population is killed but a small number of metabolically quiescent bacteria called persisters survive antibiotic treatment. The mechanism of this bacterial persistence is poorly understood. In Escherichia coli, we identified a new persistence gene, phoU, whose inactivation leads to a generalized higher susceptibility than that of the parent strain to a diverse range of antibiotics, including ampicillin, norfloxacin, and gentamicin, and stresses, such as starvation, acid pH, heat, peroxide, weak acids, and energy inhibitors, especially in stationary phase. The PhoU mutant phenotype could be complemented by a functional phoU gene. Mutation in PhoU leads to a metabolically hyperactive status of the cell, as shown by an increased expression of energy production genes, flagella, and chemotaxis genes and a defect in persister formation. PhoU, whose expression is regulated by environmental changes like nutrient availability and age of culture, is a global negative regulator beyond its role in phosphate metabolism and facilitates persister formation by the suppression of many important cellular metabolic processes. A new model of persister formation based on PhoU as a persister switch is proposed. PhoU may be an ideal drug target for designing new drugs that kill persister bacteria for more effective control of bacterial infections. PMID- 17420207 TI - Effects of carbapenem exposure on the risk for digestive tract carriage of intensive care unit-endemic carbapenem-resistant Pseudomonas aeruginosa strains in critically ill patients. AB - To determine the epidemiology and risk factors for carbapenem-resistant Pseudomonas aeruginosa (CR-PA) digestive tract colonization, weekly rectal and pharyngeal swabs were obtained in two serial incidence surveys (266 patients). Forty-two (16%) patients were CR-PA colonized (12 [29%] on admission and 30 [71%] in intensive care units). Pulsed-field gel electrophoresis showed extensive clonal diversity, although one specific clone (type B) was isolated from 11 patients. The presence of similar genotypes of CR-PA colonizing 30% of the CR-PA colonized patients suggests the occurrence of cross-colonization; in addition, 10 pairs of carbapenem-susceptible P. aeruginosa (CS-PA) and subsequent CR-PA strains isolated from the same patients were found to be clonally identical and were considered to have been endogenously acquired (33%). All endogenously acquired CR-PA strains were isolated after exposure to a carbapenem, and 80% showed a phenotype of imipenem resistance (IR pattern) alone, while 67% of the CR PA strains acquired by cross-transmission exhibited a multiresistant (MR) phenotype, with previous carbapenem exposure in 44%. Logistic regression analysis identified severity of acute illness (odds ratio [OR], 1.0; 95% confidence interval [CI], 1.0 to 1.1), prior carbapenem use (OR, 7.8; 95% CI, 1.7 to 35.3), and prior use of fluoroquinolones (OR, 11.0; 95% CI, 1.7 to 67.9) as independent risk factors for CR-PA digestive tract colonization. Overall, the local epidemiology of CR-PA digestive tract colonization was characterized by polyclonal endemicity with phenotype patterns of IR and MR divided evenly between patients. Restricting the use of particular agents, such as carbapenems and fluoroquinolones, should be considered advisable to minimize the problem of this antibiotic resistance. However, the possible risk for development of collateral unexpected bacterial resistance patterns should be accurately monitored. PMID- 17420208 TI - SME-2-producing Serratia marcescens isolate from Switzerland. PMID- 17420209 TI - Clinical response and tolerability to and safety of saquinavir with low-dose ritonavir in human immunodeficiency virus type 1-infected mothers and their infants. AB - Saquinavir boosted with low-dose ritonavir given with zidovudine and lamivudine was well tolerated by pregnant women and their infants. All mothers had <400 human immunodeficiency virus type 1 RNA copies/ml at delivery. Two had elevated liver transaminases and amylase. Seven infant adverse events were possibly treatment related (anemia, neutropenia, and hyperbilirubinemia). PMID- 17420210 TI - In vitro activities of CG400549, a novel FabI inhibitor, against recently isolated clinical staphylococcal strains in Korea. AB - The in vitro activities of CG400549, a novel FabI inhibitor, were compared to those of linezolid and commonly used antimicrobials against recent bacterial isolates. CG400549 had an MIC(90) of 0.5 microg/ml for Staphylococcus aureus strains and was more potent than either linezolid or vancomycin. PMID- 17420211 TI - Serum differentially alters the antifungal properties of echinocandin drugs. AB - Antifungal efficacies of the echinocandin drugs caspofungin, micafungin, and anidulafungin were reduced significantly in the presence of 50% human serum, which yielded nearly equivalent MICs or minimum effective concentrations against diverse Candida spp. and Aspergillus spp. Consistent with a direct drug interaction, serum decreased the sensitivity of glucan synthase to echinocandin drugs. PMID- 17420212 TI - Lysostaphin in treatment of neonatal Staphylococcus aureus infection. AB - This study describes lysostaphin's effect against methicillin-sensitive Staphylococcus aureus in suckling rats. Standard techniques determined minimal inhibitory and bactericidal concentrations, pharmacokinetics, and efficacy. The numbers of surviving rats after vancomycin, oxacillin, and lysostaphin treatment were comparable and were different from that of controls (P < 0.00001). Lysostaphin appears effective in the treatment of neonatal S. aureus infection. PMID- 17420213 TI - Identification of the novel narrow-spectrum beta-lactamase SCO-1 in Acinetobacter spp. from Argentina. AB - By studying the beta-lactamase content of several Acinetobacter spp. isolates from Argentina, producing the expanded-spectrum beta-lactamases (ESBL) VEB-1a or PER-2, a novel Ambler class A beta-lactamase gene was identified. It encoded the narrow-spectrum beta-lactamase SCO-1, whose activity was inhibited by clavulanic acid. SCO-1 hydrolyzes penicillins at a high level and cephalosporins and carbapenems at a very low level. beta-Lactamase SCO-1 was identified from unrelated VEB-1a-positive or PER-2-positive Acinetobacter spp. isolates recovered from three hospitals. The bla(SCO-1) gene was apparently located on a plasmid of ca. 150 kb from all cases but was not associated with any ESBL-encoding gene. The G+C content of the bla(SCO) gene was 52%, a value that does not correspond to that of the A. baumannii genome (39%). beta-Lactamase SCO-1 shares 47% amino acid identity with CARB-5 and ca. 40% with the enzymes TEM, SHV, and CTX-M. A gene encoding a putative resolvase was identified downstream of the bla(SCO-1) gene, but its precise way of acquisition remains to be determined. PMID- 17420214 TI - Inhibitory activities of three classes of acyclic nucleoside phosphonates against murine polyomavirus and primate simian virus 40 strains. AB - Murine polyomavirus and simian virus 40 were used to evaluate the potencies of the compounds of three classes of acyclic nucleoside phosphonates: (i) the original HPMP (3-hydroxy-2-phosphonomethoxypropyl) and PME (2 phosphonomethoxyethyl) derivatives, (ii) the 6-[2-(phosphonomethoxy)alkoxy]-2,4 diaminopyrimidine (DAPy) derivatives, and (iii) a new class of HPMP derivatives containing a 5-azacytosine moiety. The last class showed the highest activities and selectivities against both polyomaviruses. PMID- 17420215 TI - Activity of T-1106 in a hamster model of yellow Fever virus infection. AB - Yellow fever virus (YFV) causes 30,000 deaths worldwide, despite the availability of a vaccine. There are no approved antiviral therapies for the treatment of YFV disease in humans, and, therefore, these studies were designed to investigate the anti-YFV properties of T-1106, a substituted pyrazine, in a hamster model of YFV disease. Intraperitoneal (i.p.) treatment with 100 mg/kg of body weight/day of T 1106 starting 4 h prior to virus inoculation and continuing twice daily through 7 days post-virus inoculation (dpi) resulted in significantly improved survival, alanine aminotransferase levels in the serum, weight gain, and mean day to death. Virus titer in the liver at 4 dpi was significantly reduced in treated animals, as determined by both quantitative real-time PCR and infectious cell culture assay. No toxicity (weight loss or mortality) was observed at a dose of 100 mg/kg/day in sham-infected control animals. The observed minimal effective dose of T-1106 was 32 mg/kg/day administered either by oral or i.p. treatment. Therapeutic treatment was effective in significantly improving survival when T 1106 was administered beginning as late as 4 days after virus challenge with twice-daily treatment for 8 days at a dose of 100 mg/kg/day. With favorable safety, bioavailability, and postviral challenge treatment efficacy, T-1106 was effective in the treatment of disease in hamsters infected with YFV and should be further studied for potential use as a therapy for human YFV disease. PMID- 17420216 TI - Neisseria gonorrhoeae isolates with reduced susceptibility to cefixime and ceftriaxone: association with genetic polymorphisms in penA, mtrR, porB1b, and ponA. AB - The recent emergence and transmission of Neisseria gonorrhoeae isolates with reduced susceptibility to expanded-spectrum cephalosporins such as cefixime and ceftriaxone have been reported. The aim of this study was to determine the correlation of different polymorphisms in the penA, mtrR, porB1b (penB), and ponA genes of N. gonorrhoeae with reduced susceptibility to cefixime and ceftriaxone. Eighteen gonococcal isolates with reduced cefixime and ceftriaxone susceptibility (Cef(i)) and two susceptible isolates were characterized using serovar determination, antibiograms, N. gonorrhoeae multiantigen sequence typing (NG MAST), and sequencing of penA, mtrR, porB1b, and ponA alleles. For the Cef(i) isolates (n = 18), the MICs of cefixime and ceftriaxone ranged between 0.032 to 0.38 mug/ml and 0.064 to 0.125 mug/ml, respectively. These isolates were assigned five different serovars and six divergent NG-MAST sequence types. Eleven isolates (61%) with higher MICs of cefixime and ceftriaxone contained a nearly identical penA mosaic allele and previously described polymorphisms in mtrR (a single nucleotide [A] deletion in the promoter), penB (mutations in porB1b encoding loop 3 of PorB1b), and ponA (ponA1 polymorphism). The remaining seven Cef(i) isolates (39%), which had somewhat lower MICs of cefixime and ceftriaxone, contained an aspartic acid insertion (Asp-345a) in PBP 2 in conjunction with alterations of 4 to 10 amino acid residues in the C-terminal region of the transpeptidase domain of penA. In conclusion, an unambiguous association between penA mosaic alleles, in conjunction with genetic polymorphisms in mtrR, porB1b, and ponA, and greater reduced susceptibility to cefixime and ceftriaxone was identified. PMID- 17420217 TI - Tigecycline Efflux as a Mechanism for Nonsusceptibility in Acinetobacter baumannii. AB - Tigecycline has an extended spectrum of in vitro antimicrobial activities, including that against multidrug-resistant Acinetobacter. After identifying bloodstream isolates of Acinetobacter with reduced susceptibilities to tigecycline, we performed a study to assess tigecycline efflux mediated by the resistance-nodulation-division-type transporter AdeABC. After exposure of two tigecycline-nonsusceptible isolates to the efflux pump inhibitor phenyl-arginine beta-naphthylamide (PABN), a fourfold reduction in the tigecycline MIC was observed. Both tigecycline-susceptible and -nonsusceptible isolates were found to carry the gene coding for the transmembrane component of the AdeABC pump, adeB, and the two-component regulatory system comprising adeS and adeR. Previously unreported point mutations were identified in the regulatory system in tigecycline-nonsusceptible isolates. Real-time PCR identified 40-fold and 54-fold increases in adeB expression in the two tigecycline-nonsusceptible isolates compared to that in a tigecycline-susceptible isolate. In vitro exposure of a tigecycline-susceptible clinical strain to tigecycline caused a rapid rise in the MIC of tigecycline from 2 microg/ml to 24 microg/ml, which was reversible with PABN. A 25-fold increase in adeB expression was observed in a comparison between this tigecycline-susceptible isolate and its isogenic tigecycline-nonsusceptible mutant. These results indicate that an efflux-based mechanism plays a role in reduced tigecycline susceptibility in Acinetobacter. PMID- 17420218 TI - Protection against chromium (VI)-induced oxidative stress and apoptosis by Nrf2. Recruiting Nrf2 into the nucleus and disrupting the nuclear Nrf2/Keap1 association. AB - Chromium (Cr) (VI) is a major environmental toxic metal and a human carcinogen. The molecular events mediating cellular responses to Cr(VI) are not clear at present. We show that Cr(VI) potently induced apoptosis and production of reactive oxygen species (ROS) in mouse hepa1c1c7 cells in a concentration dependent manner. Mouse embryonic fibroblast cells lacking Nrf2 exhibited elevated ROS production and apoptosis, which were markedly further increased by Cr(VI), suggesting a protective role of Nrf2 against Cr(VI) toxicity. Protection by Nrf2 correlated with induction of cytoprotective genes Ho-1 and Nqo1. Induction of the genes by Cr(VI) involved inhibition of ubiquitination of Nrf2 and accumulation of Nrf2 into the nucleus. In the nucleus, treatment with Cr(VI), but not phenolic antioxidant tert-butylhydroquinone, librates Nrf2 from the Nrf2/Keap1 association and recruits Nrf2 to the antioxidant response elements (ARE) located in the enhancers of Ho-1 and Nqo1. Activation of Nrf2 by Cr(VI) was accompanied by the nuclear translocation and deubiquitination of Keap1 implicating recycling of Keap1 in Nrf2 signaling. Thus, protection against Cr(VI) toxicity involves a transcriptional signaling loop that includes activation of Nrf2 by the toxic metal, transcription of ARE-driven genes, and reduction of ROS production. PMID- 17420219 TI - Evaluating the NMDA-glutamate receptor as a site of action for toluene, in vivo. AB - Acute exposure to toluene and other volatile organic solvents results in neurotoxicity characterized by nervous system depression, cognitive and motor impairment, and alterations in visual function. In vitro, toluene disrupts the function of N-methyl-D-aspartate (NMDA)-glutamate receptors, indicating that effects on NMDA receptor function may contribute to toluene neurotoxicity. NMDA glutamate receptors are widely present in the visual system and contribute to pattern-elicited visual-evoked potentials (VEPs) in rodents, a measure that is altered by toluene exposure. The present study tested the hypothesis that effects on NMDA receptors contribute to toluene-induced alterations in pattern-elicited VEPs. Prior to examining the effects of NMDA receptor agonists and antagonists on toluene-exposed animals, a dose-range study was conducted to determine the optimal dose for NMDA (agonist) and MK801 (antagonist). Dose levels of 2.5 mg/kg NMDA and 0.1 mg/kg MK801 were selected from these initial studies. In the second study, Long-Evans rats were exposed to toluene by inhalation, and VEPs were measured during toluene exposure in the presence or absence of NMDA or MK801. Pattern-elicited VEPs were collected by exposing rats to a sinusoidal pattern modulated at a temporal frequency of 4.55 Hz. Following collection of baseline VEPs, rats were injected with either saline, NMDA (2.5 mg/kg, ip), or MK801 (0.1 mg/kg, ip) and 10 min later were exposed to air or toluene (2000 ppm). VEP amplitudes were calculated for 1x (F1) and 2x stimulus frequency (F2). The F2 amplitude was reduced by approximately 60, 60, and 50% in the toluene-exposed groups (TOL): SALINE/TOL (n = 11), NMDA/TOL (2.5 mg/kg; n = 13), and NMDA/TOL (10 mg/kg, n = 11), respectively. Thus, NMDA (2.5 and 10 mg/kg) did not significantly affect toluene-mediated F2 amplitude effects. Administration of 0.1 mg/kg MK801 prior to toluene exposure blocked the F2 amplitude decreases caused by toluene (n = 9). However, when 0.1 mg/kg MK801 was administered 20 min after the onset of toluene exposure, toluene-mediated F2 amplitude decreases persisted despite the challenge by MK801. These data support the hypothesis that acute actions of toluene on pattern-elicited VEPs involve NMDA receptors. PMID- 17420220 TI - Dysgenesis and histological changes of genitals and perturbations of gene expression in male rats after in utero exposure to antiandrogen mixtures. AB - We investigated the ability of a mixture of three androgen receptor antagonists to induce disruption of male sexual differentiation after perinatal exposure. The aim was to assess whether the joint effects of vinclozolin, flutamide, and procymidone can be predicted based on dose-response data of the individual chemicals. Chemicals were administered orally to pregnant Wistar rats from gestational day 7 to postnatal day 16. Changes in reproductive organ weights and of androgen-regulated gene expression in prostates from male rat pups were chosen as end points for extensive dose-response studies. With all end points, the joint effects of the three antiandrogens were dose additive. Histological evaluations showed that dysgenesis and hypoplasia of prostates, seminal vesicles, and epididymis were seen with the highest mixture doses. No changes were observed in any single-compound low-dose group for these lesions, nor were there histopathological changes in the testes. Pronounced dysgenesis of external genitals was observed with all doses of the mixture, and severe dysgenesis was seen with a mixture for which the individual compounds caused no effects. A combination of doses of each chemical that on its own did not produce significant reductions in the weights of seminal vesicles and PBP C3 expression induced a marked mixture effect. Thus, antiandrogens cause additive effects on end points of various molecular complexities such as alterations at the morphological and the molecular level. Exposure to antiandrogens, which appears to exert only small effects when judged on a chemical-by-chemical basis, may induce marked responses in concert with, possibly unrecognized, similarly acting chemicals. PMID- 17420221 TI - Styrene induced alterations in biomarkers of exposure and effects in the cochlea: mechanisms of hearing loss. AB - It is known that styrene is ototoxic and causes cochlear damage starting from the middle turn. However, the cellular mechanism underlying styrene ototoxicity is still unclear. In this study, rats were exposed to styrene by gavage at different doses once a day for varying periods. Styrene levels in the cochlear tissues, styrene-induced permanent hearing loss, cochlear disruptions, and cell death pathways were determined. Styrene concentration in the cochlea varied along with the basilar membrane with the lowest level in the basal turn being consistent with the lowest styrene-induced threshold shift and hair cell loss in this region. After 3 weeks of exposure (5 days per week), a dose-dependent permanent hearing loss and a hair cell loss, especially in the midfrequency region, were observed. The styrene exposure at a dose of 200 mg/kg, which induced a blood level of 6.0 +/- 1.0 microg/g, caused an average of 4.4 +/- 0.5% OHC (outer hair cell) loss and 2-5 dB threshold shift in the cochlear region of 20-70% from the apex. A significant OHC loss was not observed until 7 days of exposure at a dose of 800 mg/kg. Deiters cells appeared to be the most vulnerable target of styrene. When condensed nuclei were observed in Deiters cells after a few days of styrene exposure (800 mg/kg), other cells were still intact. Apoptotic cell death appeared to be the main cell death pathway in the cochlea after styrene exposure. In the styrene-induced apoptotic OHCs, histochemical staining detected activated caspases-9 and 8, indicating that both mitochondrial-dependent pathway and death receptor-dependent pathway were involved in the styrene-induced cell death. PMID- 17420222 TI - Uniform procedure of (1)H NMR analysis of rat urine and toxicometabonomics Part II: comparison of NMR profiles for classification of hepatotoxicity. AB - A procedure of nuclear magnetic resonance (NMR) urinalysis using pattern recognition is proposed for early detection of toxicity of investigational compounds in rats. The method is applied to detect toxicity upon administration of 13 toxic reference compounds and one nontoxic control compound (mianserine) in rats. The toxic compounds are expected to induce necrosis (bromobenzene, paracetamol, carbon tetrachloride, iproniazid, isoniazid, thioacetamide), cholestasis (alpha-naphthylisothiocyanate (ANIT), chlorpromazine, ethinylestradiol, methyltestosterone, ibuprofen), or steatosis (phenobarbital, tetracycline). Animals were treated daily for 2 or 4 days except for paracetamol and bromobenzene (1 and 2 days) and carbon tetrachloride (1 day only). Urine was collected 24 h after the first and second treatment. The animals were sacrificed 24 h after the last treatment, and NMR data were compared with liver histopathology as well as blood and urine biochemistry. Pathology and biochemistry showed marked toxicity in the liver at high doses of bromobenzene, paracetamol, carbon tetrachloride, ANIT, and ibuprofen. Thioacetamide and chlorpromazine showed less extensive changes, while the influences of iproniazid, isoniazid, phenobarbital, ethinylestradiol, and tetracycline on the toxic parameters were marginal or for methyltestosterone and mianserine negligible. NMR spectroscopy revealed significant changes upon dosing in 88 NMR biomarker signals preselected with the Procrustus Rotation method on principal component discriminant analysis (PCDA) plots. Further evaluation of the specific changes led to the identification of biomarker patterns for the specific types of liver toxicity. Comparison of our rat NMR PCDA data with histopathological changes reported in humans and/or rats suggests that rat NMR urinalysis can be used to predict hepatotoxicity. PMID- 17420223 TI - Sensitivity of (1)H NMR analysis of rat urine in relation to toxicometabonomics. Part I: dose-dependent toxic effects of bromobenzene and paracetamol. AB - (1)H nuclear magnetic resonance (NMR) spectroscopy of rat urine in combination with pattern recognition analysis was evaluated for early noninvasive detection of toxicity of investigational chemical entities. Bromobenzene (B) and paracetamol (P) were administered at five single oral dosages between 2 and 500 mg/kg and between 6 and 1800 mg/kg, respectively. The sensitivity of the proposed method to detect changes in the NMR spectra 24 and 48 h after single dosing was compared with histopathology and biochemical parameters in plasma and urine. Both B and P applied at the highest dosages induced liver necrosis and markedly increased aspartate aminotransferase (AST) and alanine aminotransferase (ALT) plasma levels. At dosages of 125 mg/kg B and 450 mg/kg P, liver necrosis and changes in AST and ALT were less pronounced, while at lower dose levels these effects could not be detected. Changes in kidney pathology or standard urine biochemistry were not observed at any of these dosages. Evaluation of the total NMR dataset showed 80 signals to be sensitive for B and P dosing. Principal component analysis on the reduced dataset revealed that NMR spectra were significantly different at dosages above 8 mg/kg (B) and 110 mg/kg (P) at both sampling times. This implies a 4- to 16-fold increased sensitivity of NMR versus histopathology and clinical chemistry in recognizing early events of liver toxicity. PMID- 17420224 TI - Stem cell science and the Lasker Award--let the science not be lost in translation. PMID- 17420225 TI - Concise review: adipose tissue-derived stromal cells--basic and clinical implications for novel cell-based therapies. AB - Compared with bone marrow-derived mesenchymal stem cells, adipose tissue-derived stromal cells (ADSC) do have an equal potential to differentiate into cells and tissues of mesodermal origin, such as adipocytes, cartilage, bone, and skeletal muscle. However, the easy and repeatable access to subcutaneous adipose tissue and the simple isolation procedures provide a clear advantage. Since extensive reviews focusing exclusively on ADSC are rare, it is the aim of this review to describe the preparation and isolation procedures for ADSC, to summarize the molecular characterization of ADSC, to describe the differentiation capacity of ADSC, and to discuss the mechanisms and future role of ADSC in cell therapy and tissue engineering. An initial effort has also been made to differentiate ADSC into hepatocytes, endocrine pancreatic cells, neurons, cardiomyocytes, hepatocytes, and endothelial/vascular cells. Whereas the lineage-specific differentiation into cells of mesodermal origin is well understood on a molecular basis, the molecular key events and transcription factors that initially allocate the ADSC to a lineage-specific differentiation are almost completely unknown. Decoding these molecular mechanisms is a prerequisite for developing novel cell therapies. PMID- 17420226 TI - CD41+/CD45+ cells without acetylcholinesterase activity are immature and a major megakaryocytic population in murine bone marrow. AB - Murine megakaryocytes (MKs) are defined by CD41/CD61 expression and acetylcholinesterase (AChE) activity; however, their stages of differentiation in bone marrow (BM) have not been fully elucidated. In murine lineage-negative (Lin( ))/CD45(+) BM cells, we found CD41(+) MKs without AChE activity (AChE(-)) except for CD41(++) MKs with AChE activity (AChE(+)), in which CD61 expression was similar to their CD41 level. Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs could differentiate into AChE(+), with an accompanying increase in CD41/CD61 during in vitro culture. Both proplatelet formation (PPF) and platelet (PLT) production for Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs were observed later than for Lin( )/CD41(++)/CD45(+)/AChE(+) MKs, whereas MK progenitors were scarcely detected in both subpopulations. GeneChip and semiquantitative polymerase chain reaction analyses revealed that the Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs are assigned at the stage between the progenitor and PPF preparation phases in respect to the many MK/PLT-specific gene expressions, including beta1-tubulin. In normal mice, the number of Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs was 100 times higher than that of AChE(+) MKs in BM. When MK destruction and consequent thrombocytopenia were caused by an antitumor agent, mitomycin-C, Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs led to an increase in AChE(+) MKs and subsequent PLT recovery with interleukin-11 administration. It was concluded that MKs in murine BM at least in part consist of immature Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs and more differentiated Lin( )/CD41(++)/CD45(+)/AChE(+) MKs. Immature Lin(-)/CD41(+)/CD45(+)/AChE(-) MKs are a major MK population compared with AChE(+) MKs in BM and play an important role in rapid PLT recovery in vivo. PMID- 17420227 TI - Genome-wide differential gene expression profiling of human bone marrow stromal cells. AB - Bone marrow stromal cells (BMSCs) reside in bone marrow and provide a lifelong source of new cells for various connective tissues. Although human BMSCs are regarded as highly suitable for the development of cell therapeutics and regenerative medicine, the molecular factors and the networks of signaling pathways responsible for their biological properties are as yet unclear. To gain a comprehensive understanding of human BMSCs at the transcriptional level, we have performed DNA microarray-based, genome-wide differential gene expression analysis with the use of peripheral blood-derived mononuclear cells (MNCs) as a baseline. The resulting molecular profile of BMSCs was revealed to share no meaningful overlap with those of other human stem cell types, suggesting that the cells might express a unique set of genes for their stemness. By contrast, the distinct molecular signature, consisting of 92 different genes whose expression strengths are at least 50-fold higher in BMSCs compared with MNCs, was shown to encompass largely a gene subset of umbilical cord blood-derived adherent cells, suggesting that adherent cells derived from bone marrow and umbilical cord blood may be defined by a common set of genes, regardless of their origin. Intriguingly, a large number of these genes, particularly ones for extracellular matrix products, coincide with normal or tumor endothelium-specific markers. Taken together, our results here provide a BMSC-specific genetic catalog that may facilitate future studies on molecular mechanisms governing core properties of these cells. PMID- 17420228 TI - IMT504, the prototype of the immunostimulatory oligonucleotides of the PyNTTTTGT class, increases the number of progenitors of mesenchymal stem cells both in vitro and in vivo: potential use in tissue repair therapy. AB - Bone marrow (BM)-derived adult mesenchymal stem cells (MSCs) have the capacity to differentiate in vitro into different cell lines. This makes them a likely source for application in tissue repair therapies. Here, we report evidence indicating that, both in vivo and in vitro, IMT504, the prototype of the PyNTTTTGT class of immunostimulatory oligonucleotides, significantly increases the number of fibroblast colony-forming units (CFU-Fs) that originate MSCs. When rat BM cells were cultured with IMT504, the mean number of CFU-Fs increased about three times as compared with untreated controls (CFU-F: 19 +/- 6.3 vs. 6.8 +/- 2.0/2 x 10(6) seeded BM cells, p = .03). Furthermore, rats inoculated with IMT504 had a significantly higher number of CFU-Fs both in BM (CFU-F: 124 +/- 33 vs. 38 +/- 17/femur, p = .04) and in peripheral blood (animals with detectable CFU-Fs in circulation 8/12 vs. 2/12, p = .04) as compared with untreated animals. On the other hand, BM-derived adherent cells either treated in vitro with IMT504 or obtained from animals injected with IMT504 possess the capacity to differentiate to the osteogenic and adipogenic cell lineages as regular MSCs. Finally, we found that repair of a bone defect was accelerated in rats injected with IMT504 as compared with control animals (area with consolidated bone: 80% +/- 6.4% vs. 49% +/- 3.5%, p = .03, n = 10 rats per group). Importantly, when two human BM were cultured in the presence of IMT504, the mean number of fibroblastic adherent colonies also increased as compared with controls. These results suggest the possibility of clinical use of IMT504 in bone, and presumably other, tissue repair therapies. PMID- 17420229 TI - Baculoviral vector-mediated transient and stable transgene expression in human embryonic stem cells. AB - Human embryonic stem (hES) cells as a renewable cell source have great prospective applications in both developmental biology research and regenerative medicine. To realize these potentials, the development of effective and safe genetic manipulation methods in hES cells is an obvious demand. We report here that baculoviral vectors were able to transduce hES cells efficiently. In transient transduction experiments, a recombinant baculoviral vector equipped with a human elongation factor 1-alpha promoter and a woodchuck hepatitis post transcriptional regulatory element transduced up to 80% of cells in hES cell clumps and embryoid bodies. For prolonged transgene expression, hybrid baculoviral vectors that have incorporated a rep gene and inverted terminal repeat sequences from adeno-associated virus were produced. These hybrid vectors yielded stable transgene expression during the prolonged undifferentiated proliferation of hES cells and after differentiation. Baculoviral transduction did not affect the normal growth, phenotype, and pluripotency of hES cells. Thus, baculoviral vectors suitable for both transient overexpression and long-term stable expression are an attractive option for genetic manipulation of hES cells. PMID- 17420230 TI - Hemopoietic stem cells with higher hemopoietic potential reside at the bone marrow endosteum. AB - It is now evident that hemopoietic stem cells (HSC) are located in close proximity to bone lining cells within the endosteum. Accordingly, it is unlikely that the traditional method for harvesting bone marrow (BM) from mice by simply flushing long bones would result in optimal recovery of HSC. With this in mind, we have developed improved methodologies based on sequential grinding and enzymatic digestion of murine bone tissue to harvest higher numbers of BM cells and HSC from the endosteal and central marrow regions. This methodology resulted in up to a sixfold greater recovery of primitive hemopoietic cells (lineage( )Sca(+)Kit(+) [LSK] cells) and HSC as shown by transplant studies. HSC from different anatomical regions of the marrow exhibited important functional differences. Compared with their central marrow counterparts, HSC isolated from the endosteal region (a) had 1.8-fold greater proliferative potential, (b) exhibited almost twofold greater ability to home to the BM following tail vein injection and to lodge in the endosteal region, and (c) demonstrated significantly greater long-term hemopoietic reconstitution potential as shown using limiting dilution competitive transplant assays. PMID- 17420231 TI - Identification of midgut and salivary glands as specific and distinct barriers to efficient tick-borne transmission of Anaplasma marginale. AB - Understanding the determinants of efficient tick-borne microbial transmission is needed to better predict the emergence of highly transmissible pathogen strains and disease outbreaks. Although the basic developmental cycle of Anaplasma and Ehrlichia spp. within the tick has been delineated, there are marked differences in the ability of specific strains to be efficiently tick transmitted. Using the highly transmissible St. Maries strain of Anaplasma marginale in Dermacentor andersoni as a positive control and two unrelated nontransmissible strains, we identified distinct barriers to efficient transmission within the tick. The Mississippi strain was unable to establish infection at the level of the midgut epithelium despite successful ingestion of infected blood following acquisition feeding on a bacteremic animal host. This inability to colonize the midgut epithelium prevented subsequent development within the salivary glands and transmission. In contrast, A. marginale subsp. centrale colonized the midgut and then the salivary glands, replicating to a titer indistinguishable from that of the highly transmissible St. Maries strain and at least 100 times greater than that previously associated with successful transmission. Nonetheless, A. marginale subsp. centrale was not transmitted, even when a large number of infected ticks was used for transmission feeding. These results establish that there are at least two specific barriers to efficient tick-borne transmission, the midgut and salivary glands, and highlight the complexity of the pathogen-tick interaction. PMID- 17420232 TI - Growth of Yersinia pseudotuberculosis in mice occurs independently of Toll-like receptor 2 expression and induction of interleukin-10. AB - Pathogenic Yersinia translocates effector proteins into target cells via a type III secretion system (TTSS), modulating the host immune response. A component of the TTSS translocon, LcrV, has been implicated in preventing inflammation through Toll-like receptor 2 (TLR2) by inducing expression of the anti-inflammatory cytokine interleukin-10 (IL-10). TLR2(-/-) mice were reported to be less susceptible to the enteropathogen Yersinia enterocolitica. To determine whether TLR2 also plays a role in recognition of the enteropathogen Yersinia pseudotuberculosis and whether this results in an immune response that is detrimental to the host, we evaluated the macrophage cytokine response to live Y. pseudotuberculosis and analyzed the susceptibility of TLR2(-/-) mice to enteropathogenic Yersinia. We find that Yersinia induction of macrophage IL-10 occurs independently of TLR2 and LcrV and is blocked by the TTSS. In particular, the TTSS effector protein YopJ, which inhibits production of the inflammatory cytokine tumor necrosis factor alpha (TNF-alpha), also inhibits IL-10 expression. Consistent with these results, IL-10 is undetectable in Y. pseudotuberculosis infected mouse tissues until advanced stages of infection. In addition, we find that TLR2(-/-) mice (derived independently from those used in previous studies) do not display altered susceptibility to enteropathogenic Yersinia compared to wild-type mice. Tissue levels of IL-10, as well as the inflammatory cytokines TNF alpha, IL-6, and gamma interferon and the chemokine macrophage chemotactic protein 1, are similar in TLR2(+/+) and TLR2(-/-) mice during enteropathogenic Yersinia infection. Therefore, the absence of TLR2 alone does not affect the cytokine response of macrophages to, or the in vivo growth and survival of, enteropathogenic Yersinia. PMID- 17420233 TI - Evading the proteasome: absence of lysine residues contributes to pertussis toxin activity by evasion of proteasome degradation. AB - Pertussis toxin (PT) is an important virulence factor produced by Bordetella pertussis. PT holotoxin comprises one enzymatically active A subunit (S1), associated with a pentamer of B subunits. PT is an ADP-ribosyltransferase that modifies several mammalian heterotrimeric G proteins. Some bacterial toxins are believed to undergo retrograde intracellular transport through the Golgi apparatus to the endoplasmic reticulum (ER). The ER-associated degradation (ERAD) pathway involves the removal of misfolded proteins from the ER and degradation upon their return to the cytosol; this pathway may be exploited by PT and other toxins. In the cytosol, ERAD substrates are ubiquitinated at lysine residues, targeting them to the proteasome for degradation. We hypothesize that S1 avoids ubiquitination and proteasome degradation due to its lack of lysine residues. We predicted that the addition of lysine residues would reduce PT toxicity by allowing ubiquitination and degradation to occur. Variant forms of PT were engineered, replacing one, two, or three arginines with lysines in a variety of locations on S1. Several variants were identified with wild-type in vitro enzymatic activity but reduced cellular activity, consistent with our hypothesis. Significant recovery of the cellular activity of these variants was observed when CHO cells were pretreated with a proteasome inhibitor. We concluded that the replacement of arginine residues with lysine in the S1 subunit of PT renders the toxin subject to proteasomal degradation, suggesting that wild-type PT avoids proteasome degradation due to an absence of lysine residues. PMID- 17420234 TI - Functional analysis of antigen 43 in uropathogenic Escherichia coli reveals a role in long-term persistence in the urinary tract. AB - Escherichia coli is the primary cause of urinary tract infection (UTI) in the developed world. The major factors associated with the virulence of uropathogenic E. coli (UPEC) are fimbrial adhesins, which mediate specific attachment to host receptors and trigger innate host responses. Another group of adhesins is represented by the autotransporter subgroup of proteins. The best characterized of these proteins, antigen 43 (Ag43), is a self-recognizing adhesin that is associated with cell aggregation and biofilm formation in E. coli K-12. The sequenced genome of prototype UPEC strain CFT073 contains two variant Ag43 encoding genes located on pathogenicity islands. The biological significance of both of these genes and their role in UPEC pathogenesis have not been investigated previously. Here we performed a detailed molecular characterization analysis of Ag43a (c3655) and Ag43b (c1273) from UPEC CFT073. Expression of Ag43a and Ag43b in a K-12 background revealed that they possess different functional properties. Ag43a produced a strong aggregation phenotype and promoted significant biofilm growth. Deletion mutants and strains constitutively expressing Ag43a and Ag43b were also constructed using CFT073. When these mutants were analyzed in a mouse model of UTI, Ag43a (but not Ag43b) promoted long-term persistence in the urinary bladder. Our findings demonstrate that Ag43a contributes to UPEC disease pathogenesis and reveal that there are pathogenicity adapted variants of Ag43 with distinct virulence-related functions. PMID- 17420235 TI - Identification of a novel two-partner secretion locus in Moraxella catarrhalis. AB - Although Moraxella catarrhalis continues to be a significant cause of disease in both children and adults, the steps involved in pathogenesis remain poorly understood. We have identified three open reading frames in the M. catarrhalis genome that encode homologues of the two-partner secretion system (TPS). The sequenced M. catarrhalis hemagglutinin-like locus of strain 7169 has a unique gene organization composed in the order of mchA1, mchB, and mchA2, where mchA1 is divergent. MchA1 and MchA2 are 74% identical at the amino acid level and diverge only in the C-terminal regions. The TPS motif identified in the common N-terminal regions of MchA1 and MchA2 was found to be homologous to the filamentous hemagglutinin of Bordetella pertussis, and MchB has homology to other TpsB transporters. The presence of MchA1 and MchA2 in outer membrane protein preparations and concentrated culture supernatants (CCSs) of strain 7169 was confirmed by immunoblotting using specific antisera. Nanoscale liquid chromatography-tandem mass spectrometry peptide sequencing of the antibody reactive bands from the CCSs was performed and demonstrated that 13 different peptides mapped to identical regions of MchA1 and MchA2. Quantitative adherence assays revealed a decrease of binding to primary normal human bronchial epithelial cells by the mch mutants 7169mchB and 7169mchA1A2B compared to that by the wild-type strain. These studies show that MchA1, MchA2, and MchB are components of a novel TPS identified in M. catarrhalis and suggest that these proteins may be involved in colonization. PMID- 17420236 TI - Host-dependent trigger of caspases and apoptosis by Legionella pneumophila. AB - The Dot/Icm system of Legionella pneumophila triggers activation of caspase-3 during early stages of infection of human macrophages, but apoptosis is delayed until late stages of infection. During early stages of infection of mouse macrophages, the organism triggers rapid caspase-1-mediated cytotoxicity, which is mediated by bacterial flagellin. However, it is not known whether caspase-1 is triggered by L. pneumophila in human macrophages or whether caspase-3 is activated in permissive or nonpermissive mouse macrophages. Using single-cell analyses, we show that the wild-type strain of L. pneumophila does not trigger caspase-1 activation throughout the intracellular infection of human monocyte derived macrophages (hMDMs), even when the flagellated bacteria escape into the cytoplasm during late stages. Using single-cell analyses, we show that the Dot/Icm system of L. pneumophila triggers caspase-3 but not caspase-1 within permissive A/J mouse bone marrow-derived primary macrophages by 2 to 8 h, but apoptosis is delayed until late stages of infection. While L. pneumophila triggers a Dot/Icm-dependent activation of caspase-1 in nonpermissive BALB/c mouse-derived macrophages, caspase-3 is not activated at any stage of infection. We show that robust intrapulmonary replication of the wild-type strain of L. pneumophila in susceptible A/J mice is associated with late-stage Dot/Icm dependent pulmonary apoptosis and alveolar inflammation. In the lungs of nonpermissive BALB/c mice, L. pneumophila does not replicate and does not trigger pulmonary apoptosis or alveolar inflammation. Thus, similar to hMDMs, L. pneumophila does not trigger caspase-1 but triggers caspase-3 activation during early and exponential replication in permissive A/J mouse-derived macrophages, and apoptosis is delayed until late stages of infection. The Dot/Icm type IV secretion system is essential for pulmonary apoptosis in the genetically susceptible A/J mice. PMID- 17420237 TI - National Institute of Allergy and Infectious Diseases bioinformatics resource centers: new assets for pathogen informatics. PMID- 17420239 TI - Interleukin-8 secretion in response to aferric enterobactin is potentiated by siderocalin. AB - Siderophores are low-molecular-weight iron chelators secreted by microbes to obtain iron under deprivation. We hypothesized that the catecholate siderophore enterobactin, produced by Enterobacteriaceae, serves as a proinflammatory signal for respiratory epithelial cells. Respiratory tract responses were explored, since at this site siderocalin, an enterobactin-binding mammalian gene product, is expressed inducibly at high levels and enterobactin-secreting respiratory flora is rare, suggesting selection against a dependence on enterobactin. Addition of aferric, but not iron-saturated, enterobactin elicits a dose dependent increase in secretion of the proinflammatory chemokine interleukin-8 by human respiratory epithelial cells in culture. This response to purified enterobactin is potentiated by recombinant siderocalin at physiologically relevant concentrations. Conditioned media from genetically modified Escherichia coli strains expressing various levels of enterobactin induce an enterobactin mediated proinflammatory response. Siderocalin has been shown to deliver enterobactin to other mammalian cell types, exogenously supplied siderocalin can be detected within epithelial cells, and siderocalin increases delivery of enterobactin to the intracellular compartment. Although many siderophores perturb labile cellular iron pools, only enterobactin elicits interleukin-8 secretion, suggesting that iron chelation is necessary but not sufficient. Thus, aferric enterobactin may be a proinflammatory signal for respiratory epithelial cells, permitting detection of microbial communities that have disturbed local iron homeostasis, and siderocalin expression by the host amplifies this signal. This may be a novel mechanism for the mucosa to respond to metabolic signals of expanding microbial communities. PMID- 17420240 TI - Genome-wide identification of Francisella tularensis virulence determinants. AB - Francisella tularensis is a gram-negative pathogen that causes life-threatening infections in humans and has potential for use as a biological weapon. The genetic basis of the F. tularensis virulence is poorly understood. This study screened a total of 3,936 transposon mutants of the live vaccine strain for infection in a mouse model of respiratory tularemia by signature-tagged mutagenesis. We identified 341 mutants attenuated for infection in the lungs. The transposon disruptions were mapped to 95 different genes, virtually all of which are also present in the genomes of other F. tularensis strains, including human pathogenic F. tularensis strain Schu S4. A small subset of these attenuated mutants carried insertions in the genes encoding previously known virulence factors, but the majority of the identified genes have not been previously linked to F. tularensis virulence. Among these are genes encoding putative membrane proteins, proteins associated with stress responses, metabolic proteins, transporter proteins, and proteins with unknown functions. Several attenuated mutants contained disruptions in a putative capsule locus which partially resembles the poly-gamma-glutamate capsule biosynthesis locus of Bacillus anthracis, the anthrax agent. Deletional mutation analysis confirmed that this locus is essential for F. tularensis virulence. PMID- 17420241 TI - Role of D-alanylation of Streptococcus gordonii lipoteichoic acid in innate and adaptive immunity. AB - In recent years, there has been considerable interest in using the oral commensal gram-positive bacterium Streptococcus gordonii as a live vaccine vector. The present study investigated the role of d-alanylation of lipoteichoic acid (LTA) in the interaction of S. gordonii with the host innate and adaptive immune responses. A mutant strain defective in d-alanylation was generated by inactivation of the dltA gene in a recombinant strain of S. gordonii (PM14) expressing a fragment of the S1 subunit of pertussis toxin. The mutant strain was found to be more susceptible to killing by polymyxin B, nisin, magainin II, and human beta defensins than the parent strain. When it was examined for binding to murine bone marrow-derived dendritic cells (DCs), the dltA mutant exhibited 200- to 400-fold less binding than the parent but similar levels of binding were shown for Toll-like receptor 2 (TLR2) knockout DCs and HEp-2 cells. In a mouse oral colonization study, the mutant showed a colonization ability similar to that of the parent and was not able to induce a significant immune response. The mutant induced significantly less interleukin 12p70 (IL-12p70) and IL-10 than the parent from DCs. LTA purified from the bacteria induced tumor necrosis factor-alpha and IL-6 production from wild-type DCs but not from TLR2 knockout DCs, and the mutant LTA induced a significantly smaller amount of these two cytokines. These results show that d-alanylation of LTA in S. gordonii plays a role in the interaction with the host immune system by contributing to the relative resistance to host defense peptides and by modulating cytokine production by DCs. PMID- 17420243 TI - Mice lacking components of adaptive immunity show increased Brucella abortus virB mutant colonization. AB - The Brucella abortus type IV secretion system (T4SS), encoded by the virB genes, is essential for survival in mononuclear phagocytes in vitro. In the mouse model, a B. abortus virB mutant was initially able to colonize the spleen at the level of the wild type for approximately 3 to 5 days, which coincided with the development of adaptive immunity. To investigate the relationship between survival in macrophages cultivated in vitro and persistence in tissues in vivo, we tested the ability of mutant mice lacking components of adaptive immunity to eliminate the virB mutant from the spleen during a mixed infection with the B. abortus wild type. Ifng(-/-) or beta(2)m(-/-) mice were able to clear the virB mutant to the same degree as control mice. However, spleens of Rag1(-/-) mice and Igh6(-/-) mice were more highly colonized by the virB mutant than control mice after 14 to 21 days, suggesting that, in these mice, there is not an absolute requirement for the T4SS to mediate persistence of B. abortus in the spleen. Macrophages isolated from Igh6(-/-) mice killed the virB mutant to the same extent as macrophages from control mice, showing that the reduced ability of these mice to clear the virB mutant from the spleen does not correlate with diminished macrophage function in vitro. These results show that in the murine model host, the T4SS is required for persistence beyond 3 to 5 days after infection and suggest that the T4SS may contribute to evasion of adaptive immune mechanisms by B. abortus. PMID- 17420242 TI - Borrelia burgdorferi binding of host complement regulator factor H is not required for efficient mammalian infection. AB - The causative agent of Lyme disease, Borrelia burgdorferi, is naturally resistant to its host's alternative pathway of complement-mediated killing. Several different borrelial outer surface proteins have been identified as being able to bind host factor H, a regulator of the alternative pathway, leading to a hypothesis that such binding is important for borrelial resistance to complement. To test this hypothesis, the development of B. burgdorferi infection was compared between factor H-deficient and wild-type mice. Factor B- and C3-deficient mice were also studied to determine the relative roles of the alternative and classical/lectin pathways in B. burgdorferi survival during mammalian infection. While it was predicted that B. burgdorferi should be impaired in its ability to infect factor H-deficient animals, quantitative analyses of bacterial loads indicated that those mice were infected at levels similar to those of wild-type and factor B- and C3-deficient mice. Ticks fed on infected factor H-deficient or wild-type mice all acquired similar numbers of bacteria. Indirect immunofluorescence analysis of B. burgdorferi acquired by feeding ticks from the blood of infected mice indicated that none of the bacteria had detectable levels of factor H on their outer surfaces, even though such bacteria express high levels of surface proteins capable of binding factor H. These findings demonstrate that the acquisition of host factor H is not essential for mammalian infection by B. burgdorferi and indicate that additional mechanisms are employed by the Lyme disease spirochete to evade complement-mediated killing. PMID- 17420244 TI - Scavenger receptor C-type lectin binds to the leukocyte cell surface glycan Lewis(x) by a novel mechanism. AB - The scavenger receptor C-type lectin (SRCL) is unique in the family of class A scavenger receptors, because in addition to binding sites for oxidized lipoproteins it also contains a C-type carbohydrate-recognition domain (CRD) that interacts with specific glycans. Both human and mouse SRCL are highly specific for the Lewis(x) trisaccharide, which is commonly found on the surfaces of leukocytes and some tumor cells. Structural analysis of the CRD of mouse SRCL in complex with Lewis(x) and mutagenesis show the basis for this specificity. The interaction between mouse SRCL and Lewis(x) is analogous to the way that selectins and DC-SIGN bind to related fucosylated glycans, but the mechanism of the interaction is novel, because it is based on a primary galactose-binding site similar to the binding site in the asialoglycoprotein receptor. Crystals of the human receptor lacking bound calcium ions reveal an alternative conformation in which a glycan ligand would be released during receptor-mediated endocytosis. PMID- 17420245 TI - Three-way stabilization of the covalent intermediate in amylomaltase, an alpha amylase-like transglycosylase. AB - Amylomaltases are glycosyl hydrolases belonging to glycoside hydrolase family 77 that are capable of the synthesis of large cyclic glucans and the disproportionation of oligosaccharides. Using protein crystallography, we have generated a flip book movie of the amylomaltase catalytic cycle in atomic detail. The structures include a covalent glycosyl enzyme intermediate and a covalent intermediate in complex with an analogue of a co-substrate and show how the structures of both enzyme and substrate respond to the changes required by the catalytic cycle as it proceeds. Notably, the catalytic nucleophile changes conformation dramatically during the reaction. Also, Gln-256 on the 250s loop is involved in orienting the substrate in the +1 site. The absence of a suitable base in the covalent intermediate structure explains the low hydrolysis activity. PMID- 17420246 TI - Nucleoplasmic calcium is required for cell proliferation. AB - Ca(2+) signals regulate cell proliferation, but the spatial and temporal specificity of these signals is unknown. Here we use selective buffers of nucleoplasmic or cytoplasmic Ca(2+) to determine that cell proliferation depends upon Ca(2+) signals within the nucleus rather than in the cytoplasm. Nuclear Ca(2+) signals stimulate cell growth rather than inhibit apoptosis and specifically permit cells to advance through early prophase. Selective buffering of nuclear but not cytoplasmic Ca(2+) signals also impairs growth of tumors in vivo. These findings reveal a major physiological and potential pathophysiological role for nucleoplasmic Ca(2+) signals and suggest that this information can be used to design novel therapeutic strategies to regulate conditions of abnormal cell growth. PMID- 17420247 TI - Roles for the human ATP-dependent Lon protease in mitochondrial DNA maintenance. AB - Human mitochondrial Lon is an ATP-powered proteolytic machine that specifically binds to single-stranded G-rich DNA and RNA in vitro. However, it is unknown whether Lon binds mitochondrial DNA (mtDNA) in living cells or functions in mtDNA integrity. Here, we demonstrate that Lon interacts with the mitochondrial genome in cultured cells using mtDNA immunoprecipitation (mIP). Lon associates with sites distributed primarily within one-half of the genome and preferentially with the control region for mtDNA replication and transcription. Bioinformatic analysis of mIP data revealed a G-rich consensus sequence. Consistent with these findings, in vitro experiments showed that the affinity of Lon for single stranded DNA oligonucleotides correlates with conformity to this consensus. To examine the role of Lon in mtDNA maintenance, cells carrying an inducible short hairpin RNA for Lon depletion were used. In control and Lon-depleted cells, mtDNA copy number was essentially the same in the presence or absence of oxidative stress. However when oxidatively stressed, control cells exhibited an increased frequency of mtDNA lesions, whereas Lon-depleted cells showed little if any mtDNA damage. This suggests that oxidative mtDNA damage is permitted when Lon is present and prevented when Lon is substantially depleted. Upon oxidative stress, mIP showed reduced Lon binding to mtDNA; however binding to the control region was unaffected. It is unlikely that oxidative modification of Lon blocks its ability to bind DNA in vivo as results show that oxidized purified Lon retains sequence-specific DNA binding. Taken together, these results demonstrate that mtDNA binding is a physiological function of Lon and that cellular levels of Lon influence sensitivity to mtDNA damage. These findings suggest roles for Lon in linking protein and mtDNA quality control. PMID- 17420248 TI - The membrane-proximal portion of CD3 epsilon associates with the serine/threonine kinase GRK2. AB - The activation of protein kinases is one of the primary mechanisms whereby T cell receptors (TCR) propagate intracellular signals. To date, the majority of kinases known to be involved in the early stages of TCR signaling are protein-tyrosine kinases such as Lck, Fyn, and ZAP-70. Here we report a constitutive association between the TCR and a serine/threonine kinase, which was mediated through the membrane-proximal portion of CD3 epsilon. Mass spectrometry analysis of CD3 epsilon-associated proteins identified G protein-coupled receptor kinase 2 (GRK2) as a candidate Ser/Thr kinase. Transient transfection assays and Western blot analysis verified the ability of GRK2 to interact with the cytoplasmic domain of CD3 epsilon within a cell. These findings are consistent with recent reports demonstrating the ability of certain G protein-coupled receptors (GPCR) and G proteins to physically associate with the alpha/beta TCR. Because GRK2 is primarily involved in arresting GPCR signals, its interaction with CD3 epsilon may provide a novel means whereby the TCR can negatively regulate signals generated through GPCRs. PMID- 17420249 TI - Phosphorylated galectin-3 mediates tumor necrosis factor-related apoptosis inducing ligand signaling by regulating phosphatase and tensin homologue deleted on chromosome 10 in human breast carcinoma cells. AB - Galectin-3 (GAL3), a beta-galactoside-binding lectin, confers chemoresistance to a wide variety of cancer cell types. It may exhibit anti- or pro-apoptotic activity depending on the nature of the stimulus. We report here that introducing phosphorylated galectin-3 (P-GAL3) into GAL3-null, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-resistant human breast carcinoma cells promotes TRAIL-induced apoptotic cell death by stimulating the phosphorylation/inactivation of the pro-apoptotic molecule Bad resulting in the inhibition of mitochondrial depolarization and the release of cytochrome c. Exposure of the transfectant cells to TRAIL leads to the recruitment of the initiator capase-8 followed by activation of the effector caspase-9, independent of cytochrome c, and subsequently the processing of the executioner caspase-3. P GAL3 and phosphatase and tensin homologue deleted on chromosome 10 (PTEN) were coordinately expressed, with concomitant dephosphorylation of Akt in TRAIL sensitive cells. In contrast, overexpression of phospho-mutant GAL3 (incapable of phosphorylation) failed to elicit similar responses. Depletion of PTEN using small interference RNAs reinstated Akt phosphorylation and conferred TRAIL resistance. In addition phosphatidylinositol 3-kinase inhibitors rendered the phospho-mutant GAL3-resistant cells sensitive to TRAIL. These findings suggest a pivotal role for P-GAL3 in promoting TRAIL sensitivity through activation of a nonclassic apoptotic pathway and identify P-GAL3 as a novel regulator of PTEN. PMID- 17420250 TI - In cultured astrocytes, p53 and MDM2 do not alter hypoxia-inducible factor-1alpha function regardless of the presence of DNA damage. AB - A principal molecular mechanism by which cells respond to hypoxia is by activation of the transcription factor hypoxia-inducible factor 1alpha (HIF 1alpha). Several studies describe a binding of p53 to HIF-1alpha in a protein complex, leading to attenuated function, half-life, and abundance of HIF-1alpha. However, these reports almost exclusively utilized transformed cell lines, and many employed transfection of p53 or HIF-1alpha plasmid constructs and/or p53 and HIF-1alpha reporter constructs as surrogates for endogenous protein activity and target expression, respectively. Thus, it remains an open and important question as to whether p53 inhibits HIF-1alpha-mediated transactivation of endogenous HIF 1alpha targets in nontransformed cells. After determining in primary astrocyte cultures the HIF-1alpha targets that were most dependent on HIF-1alpha function, we examined the effect of the loss of p53 function either alone or in combination with MDM2 on expression of these targets. Although p53 null astrocyte cultures resulted in markedly increased HIF-1alpha-dependent target expression compared with controls, this altered expression was determined to be the result of increased cell density of p53 null cultures and the accompanying acidosis, not loss of p53 protein. Although activation of p53 by DNA damage induced p53 target expression in astrocytes, it did not alter hypoxia-induced HIF-1alpha target expression. Finally, a combined loss of MDM2 and p53 did not alter HIF-1alpha target expression compared with loss of p53 alone. These data strongly suggest that p53 and MDM2 do not influence the hypoxia-induced transactivation of HIF 1alpha targets, regardless of p53 activation, in primary astrocytes. PMID- 17420251 TI - Receptor-mediated endocytosis involves tyrosine phosphorylation of cortactin. AB - Efficient internalization of cell surface receptors requires actin polymerization mediated by Arp2/3 complex and cortactin, a prominent substrate of the protein tyrosine kinase Src. However, the significance of cortactin tyrosine phosphorylation in endocytosis is unknown. We found that overexpression of a cortactin mutant deficient in tyrosine phosphorylation decreased transferrin uptake. Suppression of cortactin expression by RNA interference also reduced transferrin internalization. Such inhibition was effectively rescued by overexpressing wild-type cortactin but not a cortactin mutant deficient in tyrosine phosphorylation or a mutant with deletion of the Src homology 3 domain. Likewise, purified phosphorylation-null cortactin failed to restore the formation of clathrin-coated vesicles in a cortactin-depleted cell extract. In vitro analysis revealed that Src-mediated phosphorylation enhanced the association of cortactin with dynamin-2 in a tyrosine phosphorylation-dependent manner. Quantitative analysis demonstrated that Src enhances the affinity of cortactin for dynamin-2 by more than 3-fold. On the other hand, Src-treated dynamin-2 had no effect on its interaction with cortactin. These data indicate that Src kinase is implicated in clathrin-mediated endocytosis by phosphorylation of cortactin. PMID- 17420252 TI - Structure and function of DnaA N-terminal domains: specific sites and mechanisms in inter-DnaA interaction and in DnaB helicase loading on oriC. AB - DnaA forms a homomultimeric complex with the origin of chromosomal replication (oriC) to unwind duplex DNA. The interaction of the DnaA N terminus with the DnaB helicase is crucial for the loading of DnaB onto the unwound region. Here, we determined the DnaA N terminus structure using NMR. This region (residues 1-108) consists of a rigid region (domain I) and a flexible region (domain II). Domain I has an alpha-alpha-beta-beta-alpha-beta motif, similar to that of the K homology (KH) domain, and has weak affinity for oriC single-stranded DNA, consistent with KH domain function. A hydrophobic surface carrying Trp-6 most likely forms the interface for domain I dimerization. Glu-21 is located on the opposite surface of domain I from the Trp-6 site and is crucial for DnaB helicase loading. These findings suggest a model for DnaA homomultimer formation and DnaB helicase loading on oriC. PMID- 17420253 TI - Signaling through a G Protein-coupled receptor and its corresponding G protein follows a stoichiometrically limited model. AB - The bradykinin receptor is a G protein-coupled receptor (GPCR) that is coupled to the Galpha(q) family of heterotrimeric G proteins. In general, a GPCR can exert intracellular signals either by transiently associating with multiple diffusing G protein subunits or by activating a G protein that is stably bound to the receptor, thus generating a signal that is limited by the stoichiometry of the complex. Here we have distinguished between these models by monitoring the association of type 2 bradykinin receptor (B(2)R) and the Galpha(q)/Gbetagamma heterotrimer in living human embryonic kidney 293 cells expressing fluorescent tagged proteins. Stable B(2)R-Galpha(q) x Gbetagamma complexes are observed in resting cells by fluorescence resonance energy transfer from either Galpha(q) eCFP or eCFP-Gbetagamma to B(2)R-eYFP. Stimulating the cells with bradykinin causes detachment of B(2)R from the G protein subunits as the receptor internalizes into early endosomes, with a corresponding elimination of B(2)R-G protein fluorescence resonance energy transfer because Galpha(q) and its associated Gbetagamma remain on the plasma membrane. Single point and scanning fluorescence correlation spectroscopy measurements show that a portion of B(2)R molecules diffuses with a mobility corresponding to dimers or small oligomers, whereas a second fraction diffuses in higher order molecular assemblies. Our studies support a model in which receptors are pre-coupled with their corresponding G proteins in the basal state of cells thereby limiting the response to an external signal to a defined stoichiometry that allows for a rapid and directed cellular response. PMID- 17420254 TI - Rhizobium etli CE3 bacteroid lipopolysaccharides are structurally similar but not identical to those produced by cultured CE3 bacteria. AB - Rhizobium etli CE3 bacteroids were isolated from Phaseolus vulgaris root nodules. The lipopolysaccharide (LPS) from the bacteroids was purified and compared with the LPS from laboratory-cultured R. etli CE3 and from cultures grown in the presence of anthocyanin. Comparisons were made of the O-chain polysaccharide, the core oligosaccharide, and the lipid A. Although LPS from CE3 bacteria and bacteroids are structurally similar, it was found that bacteroid LPS had specific modifications to both the O-chain polysaccharide and lipid A portions of their LPS. Cultures grown with anthocyanin contained modifications only to the O-chain polysaccharide. The changes to the O-chain polysaccharide consisted of the addition of a single methyl group to the 2-position of a fucosyl residue in one of the five O-chain trisaccharide repeat units. This same change occurred for bacteria grown in the presence of anthocyanin. This methylation change correlated with the inability of bacteroid LPS and LPS from anthocyanin-containing cultures to bind the monoclonal antibody JIM28. The core oligosaccharide region of bacteroid LPS and from anthocyanin-grown cultures was identical to that of LPS from normal laboratory-cultured CE3. The lipid A from bacteroids consisted exclusively of a tetraacylated species compared with the presence of both tetra- and pentaacylated lipid A from laboratory cultures. Growth in the presence of anthocyanin did not affect the lipid A structure. Purified bacteroids that could resume growth were also found to be more sensitive to the cationic peptides, poly l-lysine, polymyxin-B, and melittin. PMID- 17420255 TI - c-Fms tyrosine 559 is a major mediator of M-CSF-induced proliferation of primary macrophages. AB - The molecular mechanisms by which binding of monocyte/macrophage colony stimulating factor to its receptor c-Fms promotes replication in primary macrophages are incompletely understood, as all previous studies involved overexpression of receptor mutants in transformed cells not endogenously expressing the receptor. To address this issue we retrovirally expressed, in bone marrow-derived macrophages, a chimeric receptor containing a range of tyrosine to phenylalanine mutations in the c-Fms cytoplasmic tail. We measured incorporation of bromodeoxyuridine as a marker of proliferation and phosphorylation of ERKs, Akt, and the receptor itself. Our data indicate that tyrosine 559 is the major mediator of receptor activation and cell death, intracellular signaling, and cell proliferation and that the tyrosine residues at positions 697 and 807 play lesser roles in these events. Importantly, we find that activation of the ERK and Akt pathways is necessary but not sufficient for induction of macrophage proliferation. Using specific small molecule inhibitors we find that a combination of the Src family kinase, phosphatidylinositol 3-kinase/Akt, phospholipase C, and ERK pathways mediates macrophage proliferation in response to M-CSF. PMID- 17420256 TI - M-CSF regulates the cytoskeleton via recruitment of a multimeric signaling complex to c-Fms Tyr-559/697/721. AB - M-CSF is known to induce cytoskeletal reorganization in macrophages and osteoclasts by activation of phosphatidylinositol 3-kinase (PI3K) and c-Src, but the detailed mechanisms remain unclear. We find, unexpectedly, that tyrosine (Tyr) to phenylalanine (Phe) mutation of Tyr-721, the PI3K binding site in the M CSF receptor c-Fms, fails to suppress cytoskeletal remodeling or actin ring formation. In contrast, mutation of c-Fms Tyr-559 to Phe blocks M-CSF-induced cytoskeletal reorganization by inhibiting formation of a Src Family Kinase SFK.c Cbl.PI3K complex and the downstream activation of Vav3 and Rac, two key mediators of actin remodeling. Using an add-back approach in which specific Tyr residues are reinserted into c-Fms inactivated by the absence of all seven functionally important Tyr residues, we find that Tyr-559 is necessary but not sufficient to transduce M-CSF-dependent cytoskeletal reorganization. Furthermore, this same add back approach identifies important roles for Tyr-697 and Tyr-721 in collaborating with Tyr-559 to recruit a multimeric signaling complex that can transduce signals from c-Fms to the actin cytoskeleton. PMID- 17420257 TI - Down-regulation of polysialic acid is required for efficient myelin formation. AB - Oligodendrocyte precursor cells modify the neural cell adhesion molecule (NCAM) by the attachment of polysialic acid (PSA). Upon further differentiation into mature myelinating oligodendrocytes, however, oligodendrocyte precursor cells down-regulate PSA synthesis. In order to address the question of whether this down-regulation is a necessary prerequisite for the myelination process, transgenic mice expressing the polysialyltransferase ST8SiaIV under the control of the proteolipid protein promoter were generated. In these mice, postnatal down regulation of PSA in oligodendrocytes was abolished. Most NCAM-120, the characteristic NCAM isoform in oligodendrocytes, carried PSA in the transgenic mice at all stages of postnatal development. Polysialylated NCAM-120 partially co localized with myelin basic protein and was present in purified myelin. The permanent expression of PSA-NCAM in oligodendrocytes led to a reduced myelin content in the forebrains of transgenic mice during the period of active myelination and in the adult animal. In situ hybridizations indicated a significant decrease in the number of mature oligodendrocytes in the forebrain. Thus, down-regulation of PSA during oligodendrocyte differentiation is a prerequisite for efficient myelination by mature oligodendrocytes. Furthermore, myelin of transgenic mice exhibited structural abnormalities like redundant myelin and axonal degeneration, indicating that the down-regulation of PSA is also necessary for myelin maintenance. PMID- 17420258 TI - Requirement for the dynein light chain km23-1 in a Smad2-dependent transforming growth factor-beta signaling pathway. AB - We have identified km23-1 as a novel transforming growth factor-beta (TGFbeta) receptor (TbetaR)-interacting protein that is also a light chain of the motor protein dynein (dynein light chain). Herein, we demonstrate by sucrose gradient analyses that, in the presence of TGFbeta but not in the absence, km23-1 was present in early endosomes with the TbetaRs. Further, confocal microscopy studies indicate that endogenous km23-1 was co-localized with endogenous Smad2 at early times after TGFbeta treatment, prior to Smad2 translocation to the nucleus. In addition, immunoprecipitation/blot analyses showed that TGFbeta regulated the interaction between endogenous km23-1 and endogenous Smad2 in vivo. Blockade of km23-1 using a small interfering RNA approach resulted in a reduction in both total intracellular Smad2 levels and in nuclear levels of phosphorylated Smad2 after TGFbeta treatment. This decrease was reversed by lactacystin, a specific inhibitor of the 26 S proteasome, suggesting that knockdown of km23-1 causes proteasomal degradation of phosphorylated (i.e. activated) Smad2. Blockade of km23-1 also resulted in a reduction in TGFbeta/Smad2-dependent ARE-Lux transcriptional activity, which was rescued by a km23-1 small interfering RNA resistant construct. In contrast, a reduction in TGFbeta/Smad3-dependent SBE2-Luc transcriptional activity did not occur under similar conditions. Furthermore, overexpression of the dynactin subunit dynamitin, which is known to disrupt dynein-mediated intracellular transport, blocked TGFbeta-stimulated nuclear translocation of Smad2. Collectively, our findings indicate for the first time that a dynein light chain is required for a Smad2-dependent TGFbeta signaling pathway. PMID- 17420259 TI - Differential potency of dominant negative connexin43 mutants in oculodentodigital dysplasia. AB - Oculodentodigital dysplasia (ODDD) is a congenital autosomal dominant disorder with phenotypic variability, which has been associated with mutations in the GJA1 gene encoding connexin43 (Cx43). Given that Cx43 mutants are thought to be equally co-expressed with wild-type Cx43 in ODDD patients, it is imperative to examine the consequence of these mutants in model systems that reflect this molar ratio. To that end, we used differential fluorescent protein tagging of mutant and wild-type Cx43 to quantitatively monitor the ratio of mutant/wild-type within the same putative gap junction plaques and co-immunoprecipitation to determine if the mutants interact with wild-type Cx43. Together the fluorescence-based assay was combined with patch clamp analysis to assess the dominant negative potency of Cx43 mutants. Our results revealed that the ODDD-linked Cx43 mutants, G21R and G138R, as well as amino terminus green fluorescent protein-tagged Cx43, were able to co-localize with wild-type Cx43 at the gap junction plaque-like structures and to co-immunoprecipitate with wild-type Cx43. All Cx43 mutants demonstrated dominant negative action on gap junctional conductance of wild-type Cx43 but not that of Cx32. More interestingly, these Cx43 mutants demonstrated different potencies in inhibiting the function of wild-type Cx43 with the G21R mutant being two times more potent than the G138R mutant. The potency difference in the dominant negative properties of ODDD-linked Cx43 mutants may have clinical implications for the various symptoms and disease severity observed in ODDD patients. PMID- 17420260 TI - Healing of hymenal injuries in prepubertal and adolescent girls: a descriptive study. AB - OBJECTIVE: The objective of this study was to identify the healing process and outcome of hymenal injuries in prepubertal and adolescent girls. METHODS: This multicenter, retrospective project used photographs to document the healing process and outcome of hymenal trauma that was sustained by 239 prepubertal and pubertal girls whose ages ranged from 4 months to 18 years. RESULTS: The injuries that were sustained by the 113 prepubertal girls consisted of 21 accidental or noninflicted injuries, 73 secondary to abuse, and 19 "unknown cause" injuries. All 126 pubertal adolescents were sexual assault victims. The hymenal injuries healed at various rates and except for the deeper lacerations left no evidence of the previous trauma. Abrasions and "mild" submucosal hemorrhages disappeared within 3 to 4 days, whereas "marked" hemorrhages persisted for 11 to 15 days. Only petechiae and blood blisters proved to be "markers" for determining the approximate age of an injury. Petechiae resolved within 48 hours in the prepubertal girls and 72 hours in the adolescents. A blood blister was detected at 34 days in an adolescent. As lacerations healed, their observed depth became shallower and their configuration smoothed out. Of the girls who sustained "superficial," "intermediate," or "deep" lacerations, 15 of 18 prepubertal girls had smooth and continuous appearing hymenal rims, whereas 24 of 41 adolescents' hymens had a normal, "scalloped" appearance and 30 of 34 had no disruption of continuity on healing. The final "width" of a hymenal rim was dependent on the initial depth of the laceration. No scar tissue formation was observed in either group of girls. CONCLUSIONS: The hymenal injuries healed rapidly and except for the more extensive lacerations left no evidence of a previous injury. There were no significant differences in the healing process and the outcome of the hymenal injuries in the 2 groups of girls. PMID- 17420261 TI - Long-term follow-up of 414 HIV-infected Romanian children and adolescents receiving lopinavir/ritonavir-containing highly active antiretroviral therapy. AB - BACKGROUND: There are no published reports of the long-term safety and effectiveness of highly active antiretroviral therapy for children and adolescents living in resource-limited settings or of large cohorts of HIV infected children and adolescents treated long-term (>48 weeks) with lopinavir/ritonavir-containing highly active antiretroviral therapy. OBJECTIVES: The purpose of this work was to evaluate the long-term outcomes of treatment of HIV-infected children and adolescents with lopinavir/ritonavir-containing highly active antiretroviral therapy in a resource-limited setting. METHODS: We studied an inception cohort of 414 HIV-infected children receiving lopinavir/ritonavir containing highly active antiretroviral therapy between November 2001 and August 2006 at the Romanian-American Children's Center in Constanta, Romania. The center provides comprehensive primary and HIV specialty care and treatment to all known HIV-infected children and adolescents living in Constanta. We measured safety and effectiveness by the percentage of children remaining on treatment, rates of mortality, and changes in plasma HIV RNA concentrations and CD4+ lymphocyte counts. RESULTS: The study population consisted predominantly of antiretroviral drug-experienced older children and adolescents with advanced HIV disease. Treatment was well tolerated, with 337 children (81%) remaining on therapy after a median duration of >4 years. Thirty-seven deaths occurred; the death rate compared favorably to prospectively collected historical data. The most recent on treatment plasma HIV RNA concentration was <400 copies per milliliter in 192 of 265 children tested. The mean baseline CD4+ lymphocyte count was 292 cells per microliter (n = 299); the mean change from baseline was +266 (n = 284), +317 (n = 260), +343 (n = 176), and +270 cells per microliter (n = 121) after 1, 2, 3, and 4 years of treatment, respectively. CONCLUSIONS: Highly active antiretroviral therapy can be administered safely and effectively to children and adolescents in resource-limited settings. Lopinavir/ritonavir-containing highly active antiretroviral therapy is a safe, effective, and durable treatment option for antiretroviral drug-experienced older children and adolescents with advanced HIV disease. PMID- 17420262 TI - Brugada syndrome masquerading as febrile seizures. AB - Fever can precipitate ventricular tachycardia in adults with Brugada syndrome, but such a link has not been reported in children. A 21-month-old white girl presented repeatedly with decreased conscious level and seizures during fever. During a typical episode, rapid ventricular tachycardia was documented. The resting 12-lead electrocardiogram revealed a Brugada electrocardiogram signature. Resting electrocardiograms of the asymptomatic brother and mother were normal, but fever in the mother and pharmacologic stress with ajmaline in the brother revealed Brugada electrocardiogram features. Genetic testing revealed an SCN5A mutation in the affected family members. PMID- 17420263 TI - Cis binding between inhibitory receptors and MHC class I can regulate mast cell activation. AB - Allergy is caused by immune effector cells, including mast cells and basophils. Cellular signaling that activates these effector cells is regulated by different inhibitory receptors on their surface. We show that human leukocyte immunoglobulin (Ig)-like receptor (LILR) B2 and its mouse orthologue, paired Ig like receptor (PIR)-B, constitutively associate to major histocompatibility complex (MHC) class I on the same cell surface (in cis). The IgE-mediated effector responses were augmented in beta(2)-microglobulin (beta(2)m) and PIR-B deficient mast cells. In addition, the increased cytokine production of beta(2)m deficient mast cells was not affected by the co-culture with MHC class I-positive mast cells, showing that less cis interaction between PIR-B and MHC class I on mast cells led to the increased cytokine release. Thus, the constitutive cis binding between LILRB2 or PIR-B and MHC class I has an essential role in regulating allergic responses. PMID- 17420264 TI - De novo DNA methyltransferase is essential for self-renewal, but not for differentiation, in hematopoietic stem cells. AB - DNA methylation is an epigenetic modification essential for development. The DNA methyltransferases Dnmt3a and Dnmt3b execute de novo DNA methylation in gastrulating embryos and differentiating germline cells. It has been assumed that these enzymes generally play a role in regulating cell differentiation. To test this hypothesis, we examined the role of Dnmt3a and Dnmt3b in adult stem cells. CD34(-/low), c-Kit(+), Sca-1(+), lineage marker(-) (CD34(-) KSL) cells, a fraction of mouse bone marrow cells highly enriched in hematopoietic stem cells (HSCs), expressed both Dnmt3a and Dnmt3b. Using retroviral Cre gene transduction, we conditionally disrupted Dnmt3a, Dnmt3b, or both Dnmt3a and Dnmt3b (Dnmt3a/Dnmt3b) in CD34(-) KSL cells purified from mice in which the functional domains of these genes are flanked by two loxP sites. We found that Dnmt3a and Dnmt3b function as de novo DNA methyltransferases during differentiation of hematopoietic cells. Unexpectedly, in vitro colony assays and in vivo transplantation assays showed that both myeloid and lymphoid lineage differentiation potentials were maintained in Dnmt3a-, Dnmt3b-, and Dnmt3a/Dnmt3b deficient HSCs. However, Dnmt3a/Dnmt3b-deficient HSCs, but not Dnmt3a- or Dnmt3b deficient HSCs, were incapable of long-term reconstitution in transplantation assays. These findings establish a critical role for DNA methylation by Dnmt3a and Dnmt3b in HSC self-renewal. PMID- 17420265 TI - Antiviral CD4+ memory T cells are IL-15 dependent. AB - Survival and intermittent proliferation of memory CD4(+) and CD8(+) T cells appear to be controlled by different homeostatic mechanisms. In particular, contact with interleukin (IL)-15 has a decisive influence on memory CD8(+) cells, but not memory CD4(+) cells. Past studies of memory CD4(+) cells have relied heavily on the use of naturally occurring memory phenotype (MP) cells as a surrogate for antigen (Ag)-specific memory cells. However, we show here that MP CD4(+) cells contain a prominent subset of rapidly proliferating major histocompatibility complex (MHC) II-dependent cells. In contrast, Ag-specific memory CD4 cells have a slow turnover rate and are MHC II independent. In irradiated hosts, these latter cells ignore IL-15 and expand in response to the elevated levels of IL-7 in the lymphopenic hosts. In contrast, in normal nonlymphopenic hosts where IL-7 levels are low, memory CD4 cells are heavily dependent on IL-15. Significantly, memory CD4(+) responsiveness to endogenous IL 15 reflects marked competition from other cells, especially CD8(+) and natural killer cells, and increases considerably after removal of these cells. Therefore, under normal physiological conditions, homeostasis of CD8(+) and CD4(+) memory cells is quite similar and involves IL-15 and IL-7. PMID- 17420266 TI - Enhancement and suppression of signaling by the conserved tail of IgG memory-type B cell antigen receptors. AB - Immunological memory is characterized by heightened immunoglobulin (Ig) G antibody production caused in part by enhanced plasma cell formation conferred by conserved transmembrane and cytoplasmic segments in isotype-switched IgG B cell receptors. We tested the hypothesis that the IgG tail enhances intracellular B cell antigen receptor (BCR) signaling responses to antigen by analyzing B cells from Ig transgenic mice with IgM receptors or chimeric IgMG receptors containing the IgG tail segment. The IgG tail segment enhanced intracellular calcium responses but not tyrosine or extracellular signal-related kinase (ERK) phosphorylation. Biochemical analysis and crosses to CD22-deficient mice established that IgG tail enhancement of calcium and antibody responses, as well as marginal zone B cell formation, was not due to diminished CD22 phosphorylation or inhibitory function. Microarray profiling showed no evidence for enhanced signaling by the IgG tail for calcium/calcineurin, ERK, or nuclear factor kappaB response genes and little evidence for any enhanced gene induction. Instead, almost half of the antigen-induced gene response in IgM B cells was diminished 50 90% by the IgG tail segment. These findings suggest a novel "less-is-more" hypothesis to explain how switching to IgG enhances B cell memory responses, whereby decreased BCR signaling to genes that oppose marginal zone and plasma cell differentiation enhances the formation of these key cell types. PMID- 17420267 TI - Viral antigen and extensive division maintain virus-specific CD8 T cells during chronic infection. AB - Efficient maintenance of memory CD8 T cells is central to long-term protective immunity. IL-7- and IL-15-driven homeostatic proliferation is essential for long term memory CD8 T cell persistence after acute infections. During chronic infections, however, virus-specific CD8 T cells respond poorly to these cytokines. Yet, virus-specific CD8 T cells often persist for long periods of time during chronic infections. We have addressed this apparent paradox by examining the mechanism for maintaining virus-specific CD8 T cells during chronic infection. We find that homeostatic cytokines (e.g., IL-7/15), inflammatory signals, and priming of recent thymic emigrants are not sufficient to maintain virus-specific CD8 T cells over time during chronic infection. Rather, our results demonstrate that viral peptide is required for virus-specific CD8 T cell persistence during chronic infection. Moreover, this viral antigen-dependent maintenance results in a dramatically different type of T cell division than is normally observed during memory T cell homeostasis. Rather than undergoing slow, steady homeostatic turnover during chronic viral infection, CD8 T cells undergo extensive peptide-dependent division, yet cell numbers remain relatively stable. These results indicate that antigen-specific CD8 T cell responses during persisting infection are maintained by a mechanism distinct from that after acute infection. PMID- 17420268 TI - IgG1 B cell receptor signaling is inhibited by CD22 and promotes the development of B cells whose survival is less dependent on Ig alpha/beta. AB - We describe a mouse strain in which B cell development relies either on the expression of membrane-bound immunoglobulin (Ig) gamma1 or mu heavy chains. Progenitor cells expressing gamma1 chains from the beginning generate a peripheral B cell compartment of normal size with all subsets, but a partial block is seen at the pro- to pre-B cell transition. Accordingly, gamma1-driven B cell development is disfavored in competition with developing B cells expressing a wild-type (WT) IgH locus. However, the mutant B cells display a long half-life and accumulate in the mature B cell compartment, and even though partial truncation of the Ig alpha cytoplasmic tail compromises their development, it does not affect their maintenance, as it does in WT cells. IgG1-expressing B cells showed an enhanced Ca(2+) response upon B cell receptor cross-linking, which was not due to a lack of inhibition by CD22. The enhanced Ca(2+) response was also observed in mature B cells that had been switched from IgM to IgG1 expression in vivo. Collectively, these results suggest that the gamma1 chain can exert a unique signaling function that can partially replace that of the Ig alpha/beta heterodimer in B cell maintenance and may contribute to memory B cell physiology. PMID- 17420269 TI - Thromboxane A2 is a key regulator of pathogenesis during Trypanosoma cruzi infection. AB - Chagas' disease is caused by infection with the parasite Trypanosoma cruzi. We report that infected, but not uninfected, human endothelial cells (ECs) released thromboxane A(2) (TXA(2)). Physical chromatography and liquid chromatography tandem mass spectrometry revealed that TXA(2) is the predominant eicosanoid present in all life stages of T. cruzi. Parasite-derived TXA(2) accounts for up to 90% of the circulating levels of TXA(2) in infected wild-type mice, and perturbs host physiology. Mice in which the gene for the TXA(2) receptor (TP) has been deleted, exhibited higher mortality and more severe cardiac pathology and parasitism (fourfold) than WT mice after infection. Conversely, deletion of the TXA(2) synthase gene had no effect on survival or disease severity. TP expression on somatic cells, but not cells involved in either acquired or innate immunity, was the primary determinant of disease progression. The higher intracellular parasitism observed in TP-null ECs was ablated upon restoration of TP expression. We conclude that the host response to parasite-derived TXA(2) in T. cruzi infection is possibly an important determinant of mortality and parasitism. A deeper understanding of the role of TXA(2) may result in novel therapeutic targets for a disease with limited treatment options. PMID- 17420270 TI - Jinx, an MCMV susceptibility phenotype caused by disruption of Unc13d: a mouse model of type 3 familial hemophagocytic lymphohistiocytosis. AB - Mouse cytomegalovirus (MCMV) susceptibility often results from defects of natural killer (NK) cell function. Here we describe Jinx, an N-ethyl-N-nitrosourea induced MCMV susceptibility mutation that permits unchecked proliferation of the virus, causing death. In Jinx homozygotes, activated NK cells and cytotoxic T lymphocytes (CTLs) fail to degranulate, although they retain the ability to produce cytokines, and cytokine levels are markedly elevated in the blood of infected mutant mice. Jinx was mapped to mouse chromosome 11 on a total of 246 meioses and confined to a 4.60-million basepair critical region encompassing 122 annotated genes. The phenotype was ascribed to the creation of a novel donor splice site in Unc13d, the mouse orthologue of human MUNC13-4, in which mutations cause type 3 familial hemophagocytic lymphohistiocytosis (FHL3), a fatal disease marked by massive hepatosplenomegaly, anemia, and thrombocytopenia. Jinx mice do not spontaneously develop clinical features of hemophagocytic lymphohistiocytosis (HLH), but do so when infected with lymphocytic choriomeningitis virus, exhibiting hyperactivation of CTLs and antigen-presenting cells, and inadequate restriction of viral proliferation. In contrast, neither Listeria monocytogenes nor MCMV induces the syndrome. In mice, the HLH phenotype is conditional, which suggests the existence of a specific infectious trigger of FHL3 in humans. PMID- 17420271 TI - The discovery value of "Big Science". AB - The increasing complexity of biomedical research is leading to the exploration of new models for large-scale collaborative research. This Big Science approach, however, has created anxieties and potential tensions between investigator-driven research, and research guided by a more organized, collaborative effort. Another potential tension exists between research conducted purely in search of new knowledge and research aimed at finding solutions. We argue that big biomedicine- the work of coordinated multidisciplinary groups that use the latest technologies to solve complex problems--can be an important way to harness the creativity of individual investigators, stimulate innovation, and supply the infrastructure, experimental systems, and resources needed to solve the urgent health problems confronted by our global society. We discuss this using the example of the Global HIV Vaccine Enterprise. PMID- 17420272 TI - LAT and NTAL mediate immunoglobulin E-induced sustained extracellular signal regulated kinase activation critical for mast cell survival. AB - Immunoglobulin E (IgE) induces mast cell survival in the absence of antigen (Ag) through the high-affinity IgE receptor, Fcepsilon receptor I (FcepsilonRI). Although we have shown that protein tyrosine kinase Syk and sustained extracellular signal-regulated kinase (Erk) activation are required for IgE induced mast cell survival, how Syk couples with sustained Erk activation is still unclear. Here, we report that the transmembrane adaptors LAT and NTAL are phosphorylated slowly upon IgE stimulation and that sustained but not transient Erk activation induced by IgE was inhibited in LAT(-/-) NTAL(-/-) bone marrow derived mast cells (BMMCs). IgE-induced survival requires Ras activation, and both were impaired in LAT(-/-) NTAL(-/-) BMMCs. Sos was preferentially required for FcepsilonRI signals by IgE rather than IgE plus Ag. Survival impaired in LAT( /-) NTAL(-/-) BMMCs was restored to levels comparable to those of the wild type by membrane-targeted Sos, which bypasses the Grb2-mediated membrane recruitment of Sos. The IgE-induced survival of BMMCs lacking Gads, an adaptor critical for the formation of the LAT-SLP-76-phospholipase Cgamma (PLCgamma) complex, was observed to be normal. IgE stimulation induced the membrane retention of Grb2 green fluorescent protein fusion proteins in wild-type but not LAT(-/-) NTAL(-/-) BMMCs. These results suggest that LAT and NTAL contribute to the maintenance of Erk activation and survival through the membrane retention of the Ras-activating complex Grb2-Sos and, further, that the LAT-Gads-SLP-76-PLCgamma and LAT/NTAL Grb2-Sos pathways are differentially required for degranulation and survival, respectively. PMID- 17420273 TI - CDK4 and CDK6 delay senescence by kinase-dependent and p16INK4a-independent mechanisms. AB - Replicative senescence of human diploid fibroblasts (HDFs) is largely implemented by the cyclin-dependent kinase (CDK) inhibitors p16(INK4a) and p21(CIP1). Their accumulation results in a loss of CDK2 activity, and cells arrest with the retinoblastoma protein (pRb) in its hypophosphorylated state. It has become standard practice to bypass the effects of p16(INK4a) by overexpressing CDK4 or a variant form that is unable to bind to INK4 proteins. Although CDK4 and CDK6 and their INK4-insensitive variants can extend the life span of HDFs, they also cause a substantial increase in the levels of endogenous p16(INK4a). Here we show that CDK4 and CDK6 can extend the life span of HDFs that have inactivating mutations in both alleles of INK4a or in which INK4a levels are repressed, indicating that overexpression of CDK4/6 is not equivalent to ablation of p16(INK4a). However, catalytically inactive versions of these kinases are unable to extend the replicative life span, suggesting that the impact of ectopic CDK4/6 depends on their ability to phosphorylate as yet unidentified substrates rather than to sequester CDK inhibitors. Since p16(INK4a) deficiency, CDK4 expression, and p53 or p21(CIP1) ablation have additive effects on replicative life span, our results underscore the idea that senescence is an integrated response to diverse signals. PMID- 17420274 TI - The three-dimensional structure of human interphase chromosomes is related to the transcriptome map. AB - The three-dimensional (3D) organization of the chromosomal fiber in the human interphase nucleus is an important but poorly understood aspect of gene regulation. Here we quantitatively analyze and compare the 3D structures of two types of genomic domains as defined by the human transcriptome map. While ridges are gene dense and show high expression levels, antiridges, on the other hand, are gene poor and carry genes that are expressed at low levels. We show that ridges are in general less condensed, more irregularly shaped, and located more closely to the nuclear center than antiridges. Six human cell lines that display different gene expression patterns and karyotypes share these structural parameters of chromatin. This shows that the chromatin structures of these two types of genomic domains are largely independent of tissue-specific variations in gene expression and differentiation state. Moreover, we show that there is remarkably little intermingling of chromatin from different parts of the same chromosome in a chromosome territory, neither from adjacent nor from distant parts. This suggests that the chromosomal fiber has a compact structure that sterically suppresses intermingling. Together, our results reveal novel general aspects of 3D chromosome architecture that are related to genome structure and function. PMID- 17420275 TI - GATA-1 and Gfi-1B interplay to regulate Bcl-xL transcription. AB - The induction of Bcl-x(L) is critical for the survival of late proerythroblasts. The erythroid-specific transcriptional network that regulates Bcl-x(L) expression in erythropoiesis remains unclear. The activation of the central erythropoietic transcriptional factor, GATA-1, leads to the early, transient induction of a transcription repressor, Gfi-1B, followed by the late induction of Bcl-x(L) during erythroid maturation in G1ER cells. Chromatin immunoprecipitation assays demonstrated that a constant level of GATA-1 binds to the Bcl-x promoter throughout the entire induction period, while Gfi-1B is transiently associated with the promoter in the early phase. The sustained expression of Gfi-1B abolished GATA-1-induced Bcl-x(L) expression. Here, we present evidence that GATA 1 binds to the noncanonical GATT motif of the Bcl-x promoter for trans activation. Gfi-1B expressed at increased levels is recruited to the Bcl-x promoter through its association with GATA-1, suppressing Bcl-x(L) transcription. Therefore, the down-regulation of Gfi-1B in the late phase of erythroid maturation is necessary for Bcl-x(L) induction. Furthermore, we show that the inhibition of Bcr-Abl kinase by treatment with imatinib caused the up-regulation of Gfi-1B in K562 cells, where Gfi-1B also cooperated with GATA-1 to repress Bcl x(L) transcription. Gfi-1B knockdown by RNA interference diminished imatinib induced apoptosis, while the overexpression of Gfi-1B sensitized K562 cells to arsenic-induced death. These findings illuminate the role of Gfi-1B in GATA-1 mediated transcription in the survival aspect of erythroid cells. PMID- 17420276 TI - N-glycolylneuraminic acid deficiency in mice: implications for human biology and evolution. AB - Humans and chimpanzees share >99% identity in most proteins. One rare difference is a human-specific inactivating deletion in the CMAH gene, which determines biosynthesis of the sialic acid N-glycolylneuraminic acid (Neu5Gc). Since Neu5Gc is prominent on most chimpanzee cell surfaces, this mutation could have affected multiple systems. However, Neu5Gc is found in human cancers and fetuses and in trace amounts in normal human tissues, suggesting an alternate biosynthetic pathway. We inactivated the mouse Cmah gene and studied the in vivo consequences. There was no evidence for an alternate pathway in normal, fetal, or malignant tissue. Rather, null fetuses accumulated Neu5Gc from heterozygous mothers and dietary Neu5Gc was incorporated into oncogene-induced tumors. As with humans, there were accumulation of the precursor N-acetylneuraminic acid and increases in sialic acid O acetylation. Null mice showed other abnormalities reminiscent of the human condition. Adult mice showed a diminished acoustic startle response and required higher acoustic stimuli to increase responses above the baseline level. In this regard, histological abnormalities of the inner ear occurred in older mice, which had impaired hearing. Adult animals also showed delayed skin wound healing. Loss of Neu5Gc in hominid ancestors approximately 2 to 3 million years ago likely had immediate and long-term consequences for human biology. PMID- 17420277 TI - Wilms' tumor protein Wt1 is an activator of the anti-Mullerian hormone receptor gene Amhr2. AB - The Wilms' tumor protein Wt1 plays an essential role in mammalian urogenital development. WT1 mutations in humans lead to a variety of disorders, including Wilms' tumor, a pediatric kidney cancer, as well as Frasier and Denys-Drash syndromes. Phenotypic anomalies in Denys-Drash syndrome include pseudohermaphroditism and sex reversal in extreme cases. We have used cDNA microarray analyses on Wt1 knockout mice to identify Wt1-dependent genes involved in sexual development. The gene most dramatically affected by Wt1 inactivation was Amhr2, encoding the anti-Mullerian hormone (Amh) receptor 2. Amhr2 is an essential factor for the regression of the Mullerian duct in males, and mutations in AMHR2 lead to the persistent Mullerian duct syndrome, a rare form of male pseudohermaphroditism. Here we show that Wt1 and Amhr2 are coexpressed during urogenital development and that the Wt1 protein binds to the promoter region of the Amhr2 gene. Inactivation and overexpression of Wt1 in cell lines was followed by immediate changes of Amhr2 expression. The identification of Amhr2 as a Wt1 target provides new insights into the role of Wt1 in sexual differentiation and indicates, in addition to its function in early gonad development and sex determination, a novel function for Wt1, namely, in Mullerian duct regression. PMID- 17420278 TI - DNA structure-induced recruitment and activation of the Fanconi anemia pathway protein FANCD2. AB - The Fanconi anemia (FA) pathway proteins are thought to be involved in the repair of irregular DNA structures including those encountered by the moving replication fork. However, the nature of the DNA structures that recruit and activate the FA proteins is not known. Because FA proteins function within an extended network of proteins, some of which are still unknown, we recently established cell-free assays in Xenopus laevis egg extracts to deconstruct the FA pathway in a fully replication-competent context. Here we show that the central FA pathway protein, xFANCD2, is monoubiquitinated (xFANCD2-L) rapidly in the presence of linear and branched double-stranded DNA (dsDNA) structures but not single-stranded or Y shaped DNA. xFANCD2-L associates with dsDNA structures in an FA core complex dependent manner but independently of xATRIP, the regulatory subunit of xATR. Formation of xFANCD2-L is also triggered in response to circular dsDNA, suggesting that dsDNA ends are not required to trigger monoubiquitination of FANCD2. The induction of xFANCD2-L in response to circular dsDNA is replication and checkpoint independent. Our results provide new evidence that the FA pathway discriminates among DNA structures and demonstrate that triggering the FA pathway can be uncoupled from DNA replication and ATRIP-dependent activation. PMID- 17420279 TI - Leptin stimulates fatty acid oxidation and peroxisome proliferator-activated receptor alpha gene expression in mouse C2C12 myoblasts by changing the subcellular localization of the alpha2 form of AMP-activated protein kinase. AB - Leptin stimulates fatty acid oxidation in skeletal muscle through the activation of AMP-activated protein kinase (AMPK) and the induction of gene expression, such as that for peroxisome proliferator-activated receptor alpha (PPARalpha). We now show that leptin stimulates fatty acid oxidation and PPARalpha gene expression in the C2C12 muscle cell line through the activation of AMPK containing the alpha2 subunit (alpha2AMPK) and through changes in the subcellular localization of this enzyme. Activated alpha2AMPK containing the beta1 subunit was shown to be retained in the cytoplasm, where it phosphorylated acetyl coenzyme A carboxylase and thereby stimulated fatty acid oxidation. In contrast, alpha2AMPK containing the beta2 subunit transiently increased fatty acid oxidation but underwent rapid translocation to the nucleus, where it induced PPARalpha gene transcription. A nuclear localization signal and Thr(172) phosphorylation of alpha2 were found to be essential for nuclear translocation of alpha2AMPK, whereas the myristoylation of beta1 anchors alpha2AMPK in the cytoplasm. The prevention of alpha2AMPK activation and the change in its subcellular localization inhibited the metabolic effects of leptin. Our data thus suggest that the activation of and changes in the subcellular localization of alpha2AMPK are required for leptin-induced stimulation of fatty acid oxidation and PPARalpha gene expression in muscle cells. PMID- 17420280 TI - Alpha11 beta1 integrin-dependent regulation of periodontal ligament function in the erupting mouse incisor. AB - The fibroblast integrin alpha11beta1 is a key receptor for fibrillar collagens. To study the potential function of alpha11 in vivo, we generated a null allele of the alpha11 gene. Integrin alpha11(-/-) mice are viable and fertile but display dwarfism with increased mortality, most probably due to severely defective incisors. Mutant incisors are characterized by disorganized periodontal ligaments, whereas molar ligaments appear normal. The primary defect in the incisor ligament leads to halted tooth eruption. alpha11beta1-defective embryonic fibroblasts displayed severe defects in vitro, characterized by (i) greatly reduced cell adhesion and spreading on collagen I, (ii) reduced ability to retract collagen lattices, and (iii) reduced cell proliferation. Analysis of matrix metalloproteinase in vitro and in vivo revealed disturbed MMP13 and MMP14 synthesis in alpha11(-/-) cells. We show that alpha11beta1 is the major receptor for collagen I on mouse embryonic fibroblasts and suggest that alpha11beta1 integrin is specifically required on periodontal ligament fibroblasts for cell migration and collagen reorganization to help generate the forces needed for axial tooth movement. Our data show a unique role for alpha11beta1 integrin during tooth eruption. PMID- 17420281 TI - Characterization of mammalian Par 6 as a dual-location protein. AB - Par 6 acts as a scaffold protein to facilitate atypical protein kinase C-mediated phosphorylation of cytoplasmic protein complexes, leading to epithelial and neuronal cell polarization. In addition to its location in the cytoplasm, Par 6 is localized to the nucleus. However, its organization and potential functions in the nucleus have not been examined. Using an affinity-purified Par 6 antibody and a chimera of Par 6 and green fluorescent protein, we show that Par 6 localizes precisely to nuclear speckles, but not to other nuclear structures, and displays characteristics of speckle proteins. We show that Par 6 colocalizes in the nucleus with Tax, a transcriptional activator of the human T-cell leukemia virus type 1 long terminal repeat, but multiple lines of evidence show that Par 6 is not directly involved in known functions of speckle proteins, including general transcription, splicing, or mRNA transport. Significantly, however, the structure of nuclear speckles is lost when Par 6 levels are reduced by Par 6-specific small interfering RNA. Therefore, we hypothesize that Par 6 in the nucleus acts as a scaffolding protein in nuclear speckle complexes, similar to its role in the cytoplasm. PMID- 17420282 TI - Pharmacokinetic parameters of tibolone and metabolites in plasma, urine, feces, and bile from ovariectomized cynomolgus monkeys after a single dose or multiple doses of tibolone. AB - Levels of nonsulfated and sulfated tibolone metabolites were determined in plasma, urine, and feces from six ovariectomized, mature female cynomolgus monkeys after a single dose and multiple p.o. doses (including bile) of tibolone using validated gas chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry assays. In plasma, the predominant nonsulfated metabolite after single and multiple dosing was the estrogenic 3alpha hydroxytibolone; levels of the estrogenic 3beta-hydroxytibolone were 10-fold lower and of progestagenic/androgenic Delta(4)-tibolone, 5-fold lower. Tibolone was undetectable. The predominant sulfated metabolite was 3alphaS,17betaS tibolone; levels of 3betaS,17betaS-tibolone were about 2-fold lower, and monosulfated 3-hydroxymetabolites were about 10-fold lower. After multiple doses, areas under the curve of nonsulfated metabolites were lower (2-fold), and those of sulfated metabolites were 25% higher. In plasma, >95% metabolites were disulfated. In urine, levels of all the metabolites after single and multiple doses were low. After a single dose, high levels of 3beta-hydroxytibolone and the 3-monosulfated metabolites (3betaS,17betaOH-tibolone and 3alphaS,17betaOH tibolone) were found in feces. After multiple dosing, 3alpha-hydroxytibolone increased, and the ratio of 3alpha/3beta-hydroxytibolone became about 1. The predominant sulfated metabolite was 3alphaS,17betaS-tibolone. Levels of all the metabolites in feces were higher after multiple doses than after a single dose. Levels of nonsulfated and 3-monosulfated metabolites were higher in feces than in plasma. Bile contained very high metabolite levels, except monosulfates. This may contribute to the metabolite content of the feces after multiple doses. 3beta Hydroxytibolone and 3alphaS,17betaS-tibolone predominated. In conclusion, tibolone had different metabolite patterns in plasma, urine, feces, and bile in monkeys. The bile contributed to the metabolite pattern in feces after multiple doses. The major excretion route was in feces. PMID- 17420283 TI - Selective tissue distribution of tibolone metabolites in mature ovariectomized female cynomolgus monkeys after multiple doses of tibolone. AB - Tibolone is a selective tissue estrogenic activity regulator (STEAR). In postmenopausal women, it acts as an estrogen on brain, vagina, and bone, but not on endometrium and breast. Despite ample supporting in vitro data for tissue selective actions, confirmative tissue levels of tibolone metabolites are not available. Therefore, we analyzed tibolone and metabolites in plasma and tissues from six ovariectomized cynomolgus monkeys that received tibolone (0.5 mg/kg/day by gavage) for 36 days and were necropsied at 1, 1.25, 2.25, 4, 6, and 24 h after the final dose. The plasma and tissue levels of active, nonsulfated (tibolone, 3alpha-hydroxytibolone, 3beta-hydroxytibolone, and Delta(4)-tibolone), monosulfated (3alpha-sulfate,17beta-hydroxytibolone and 3beta-sulfate,17beta hydroxytibolone), and disulfated (3alpha,17beta-disulfated-tibolone and 3beta,17betaS-disulfated-tibolone) metabolites were measured by validated gas chromatography with mass spectrometry and liquid chromatography with tandem mass spectrometry. Detection limits were 0.1 to 0.5 ng/ml (plasma) and 0.5 to 2 ng/g (tissues). In brain tissues, estrogenic 3alpha-hydroxytibolone was predominant with 3 to 8 times higher levels than in plasma; levels of sulfated metabolites were low. In vaginal tissues, major nonsulfated metabolites were 3alpha hydroxytibolone and the androgenic/progestagenic Delta(4)-tibolone; disulfated metabolites were predominant. Remarkably high levels of monosulfated metabolites were found in the proximal vagina. In endometrium, myometrium, and mammary glands, levels of 3-hydroxymetabolites were low and those of sulfated metabolites were high (about 98% disulfated). Delta(4)-Tibolone/3-hydroxytibolone ratios were 2 to 3 in endometrium, about equal in breast and proximal vagina, and 0.1 in plasma and brain. It is concluded that tibolone metabolites show a unique tissue specific distribution pattern explaining the tissue effects in monkeys and the clinical effects in postmenopausal women. PMID- 17420284 TI - Haploinsufficiency of EGR1, a candidate gene in the del(5q), leads to the development of myeloid disorders. AB - Loss of a whole chromosome 5 or a deletion of the long arm, del(5q), is a recurring abnormality in myelodysplastic syndromes (MDSs) and acute myeloid leukemia (AML). To identify a leukemia-related gene on chromosome 5, we previously delineated a 970-kb segment of 5q31 that is deleted in all patients examined, and prepared a transcript map of this region. EGR1 is a candidate tumor suppressor gene within the commonly deleted segment of 5q, and encodes a zinc finger transcription factor. To test the hypothesis that loss of function of Egr1 is an initiating event in the pathogenesis of AML/MDS, Egr1-deficient mice were treated with a potent DNA alkylating agent, N-ethyl-nitrosourea (ENU), to induce secondary cooperating mutations. Egr1(+/-) and Egr1(-/-) mice treated with ENU developed immature T-cell lymphomas (CD4(+), CD8(+)) or a myeloproliferative disorder (MPD) at increased rates and with shorter latencies than that of wild type littermates. The MPD was characterized by an elevated white blood cell count, anemia, and thrombocytopenia with ineffective erythropoiesis. Biallelic mutations of Egr1 were not observed in MPDs in Egr1(+/-) mice. Our data suggest that haploinsufficiency for Egr1 plays a role in murine leukemogenesis, and in the development of AML/MDS characterized by abnormalities of chromosome 5. PMID- 17420285 TI - RNAi screen identifies UBE2D3 as a mediator of all-trans retinoic acid-induced cell growth arrest in human acute promyelocytic NB4 cells. AB - All-trans retinoic acid (ATRA) has been widely used in differentiation therapy for acute promyelocytic leukemia (APL). ATRA binds to retinoic acid receptor (RAR) and triggers the formation of the transcription coactivator complex, which leads to changes in gene expression, APL cell-cycle arrest and differentiation, and clinical remission. The mechanisms responsible for ATRA's beneficial effects are still ill-defined. Here, we conducted a large-scale, unbiased short hairpin RNA (shRNA) screen aiming to identify mediators of ATRA-induced differentiation and growth arrest of APL cells. Twenty-six proteins were identified. They cover a wide range of cellular functions, including gene expression, intracellular signaling, cell death control, stress responses, and metabolic regulation, indicating the complexity of ATRA-induced cell growth control and differentiation in APL. One of these proteins, the ubiquitin-conjugating enzyme UBE2D3, is up regulated in ATRA-treated acute promyelocytic NB4 cells. UBE2D3 is physically associated with cyclin D1 and mediates ATRA-induced cyclin D1 degradation. Knocking down UBE2D3 by RNA interference (RNAi) leads to blockage of ATRA-induced cyclin D1 degradation and cell-cycle arrest. Thus, our results highlight the involvement of the ubiquitin-mediated proteolysis pathway in ATRA-induced cell cycle arrest and provide a novel strategy for modulating ATRA-elicited cellular effects. PMID- 17420286 TI - Identification of heat shock protein 32 (Hsp32) as a novel survival factor and therapeutic target in neoplastic mast cells. AB - Systemic mastocytosis (SM) is a myeloid neoplasm characterized by increased survival and accumulation of neoplastic mast cells (MCs). In most patients, the D816V-mutated variant of KIT is detectable. We report here that heat shock protein 32 (Hsp32), also known as heme oxygenase-1 (HO-1), is a novel KIT inducible survival factor in neoplastic MCs. As assessed by reverse transcription polymerase chain reaction (RT-PCR), immunocytochemistry, and Western blotting, the KIT D816V(+) MC line HMC-1.2 as well as highly enriched primary neoplastic MCs were found to express Hsp32 mRNA and the Hsp32 protein. Moreover, KIT D816V and stem cell factor (SCF)-activated wild-type KIT were found to induce Hsp32 promoter activity, expression of Hsp32 mRNA, and expression of the Hsp32 protein in Ba/F3 cells. Correspondingly, the KIT D816V-targeting drug PKC412 decreased the expression of Hsp32 as well as proliferation/survival in neoplastic MCs. The inhibitory effects of PKC412 on the survival of HMC-1.2 cells were counteracted by the HO-1 inductor hemin or lentiviral-transduced HO-1. Moreover, 2 Hsp32 targeting drugs, pegylated zinc protoporphyrin (PEG-ZnPP) and styrene maleic acid copolymer micelle-encapsulated ZnPP (SMA-ZnPP), were found to inhibit proliferation and to induce apoptosis in neoplastic MCs. Furthermore, both drugs were found to cooperate with PKC412 in producing growth inhibition. Together, these data show that Hsp32 is an important survival factor and interesting new therapeutic target in neoplastic MCs. PMID- 17420287 TI - VEGFR-1 (FLT-1), beta1 integrin, and hERG K+ channel for a macromolecular signaling complex in acute myeloid leukemia: role in cell migration and clinical outcome. AB - Leukemia cell motility and transendothelial migration into extramedullary sites are regulated by angiogenic factors and are considered unfavorable prognostic factors in acute leukemias. We have studied cross talk among (1) the vascular endothelial growth factor receptor-1, FLT-1; (2) the human eag-related gene 1 (hERG1) K(+) channels; and (3) integrin receptors in acute myeloid leukemia (AML) cells. FLT-1, hERG1, and the beta(1) integrin were found to form a macromolecular signaling complex. The latter mostly recruited the hERG1B isoform of hERG1 channels, and its assembly was necessary for FLT-1 signaling activation and AML cell migration. Both effects were inhibited when hERG1 channels were specifically blocked. A FLT-1/hERG1/beta(1) complex was also observed in primary AML blasts, obtained from a population of human patients. The co-expression of FLT-1 and hERG1 conferred a pro-migratory phenotype to AML blasts. Such a phenotype was also observed in vivo. The hERG1-positive blasts were more efficient in invading the peripheral circulation and the extramedullary sites after engraftment into immunodeficient mice. Moreover, hERG1 expression in leukemia patients correlated with a higher probability of relapse and shorter survival periods. We conclude that in AML, hERG1 channels mediate the FLT-1-dependent cell migration and invasion, and hence confer a greater malignancy. PMID- 17420288 TI - Error bars in experimental biology. AB - Error bars commonly appear in figures in publications, but experimental biologists are often unsure how they should be used and interpreted. In this article we illustrate some basic features of error bars and explain how they can help communicate data and assist correct interpretation. Error bars may show confidence intervals, standard errors, standard deviations, or other quantities. Different types of error bars give quite different information, and so figure legends must make clear what error bars represent. We suggest eight simple rules to assist with effective use and interpretation of error bars. PMID- 17420289 TI - The ubiquitin E3 ligase POSH regulates calcium homeostasis through spatial control of Herp. AB - The ubiquitin (Ub) domain protein Herp plays a crucial role in the maintenance of calcium homeostasis during endoplasmic reticulum (ER) stress. We now show that Herp is a substrate as well as an activator of the E3 Ub ligase POSH. Herp mediated POSH activation requires the Ubl domain and exclusively promotes lysine 63-linked polyubiquitination. Confocal microscopy demonstrates that Herp resides mostly in the trans-Golgi network, but, shortly after calcium perturbation by thapsigargin (Tpg), it appears mainly in the ER. Substitution of all lysine residues within the Ubl domain abolishes lysine-63-linked polyubiquitination of Herp in vitro and calcium-induced Herp relocalization that is also abrogated by the overexpression of a dominant-negative POSHV14A. A correlation exists between the kinetics of Tpg-induced Herp relocalization and POSH-dependent polyubiquitination. Finally, the overexpression of POSH attenuates, whereas the inhibition of POSH by the expression of POSHV14A or by RNA interference enhances Tpg-induced calcium burst. Altogether, these results establish a critical role for POSH-mediated ubiquitination in the maintenance of calcium homeostasis through the spatial control of Herp. PMID- 17420290 TI - Compartmentalization of androgen receptor protein-protein interactions in living cells. AB - Steroid receptors regulate gene expression in a ligand-dependent manner by binding specific DNA sequences. Ligand binding also changes the conformation of the ligand binding domain (LBD), allowing interaction with coregulators via LxxLL motifs. Androgen receptors (ARs) preferentially interact with coregulators containing LxxLL-related FxxLF motifs. The AR is regulated at an extra level by interaction of an FQNLF motif in the N-terminal domain with the C-terminal LBD (N/C interaction). Although it is generally recognized that AR coregulator and N/C interactions are essential for transcription regulation, their spatiotemporal organization is largely unknown. We performed simultaneous fluorescence resonance energy transfer and fluorescence redistribution after photobleaching measurements in living cells expressing ARs double tagged with yellow and cyan fluorescent proteins. We provide evidence that AR N/C interactions occur predominantly when ARs are mobile, possibly to prevent unfavorable or untimely cofactor interactions. N/C interactions are largely lost when AR transiently binds to DNA, predominantly in foci partly overlapping transcription sites. AR coregulator interactions occur preferentially when ARs are bound to DNA. PMID- 17420291 TI - A role for cell sex in stem cell-mediated skeletal muscle regeneration: female cells have higher muscle regeneration efficiency. AB - We have shown that muscle-derived stem cells (MDSCs) transplanted into dystrophic (mdx) mice efficiently regenerate skeletal muscle. However, MDSC populations exhibit heterogeneity in marker profiles and variability in regeneration abilities. We show here that cell sex is a variable that considerably influences MDSCs' regeneration abilities. We found that the female MDSCs (F-MDSCs) regenerated skeletal muscle more efficiently. Despite using additional isolation techniques and cell cloning, we could not obtain a male subfraction with a regeneration capacity similar to that of their female counterparts. Rather than being directly hormonal or caused by host immune response, this difference in MDSCs' regeneration potential may arise from innate sex-related differences in the cells' stress responses. In comparison with F-MDSCs, male MDSCs have increased differentiation after exposure to oxidative stress induced by hydrogen peroxide, which may lead to in vivo donor cell depletion, and a proliferative advantage for F-MDSCs that eventually increases muscle regeneration. These findings should persuade researchers to report cell sex, which is a largely unexplored variable, and consider the implications of relying on cells of one sex. PMID- 17420292 TI - Evidence for a stem cell hierarchy in the adult human breast. AB - Cellular pathways that contribute to adult human mammary gland architecture and lineages have not been previously described. In this study, we identify a candidate stem cell niche in ducts and zones containing progenitor cells in lobules. Putative stem cells residing in ducts were essentially quiescent, whereas the progenitor cells in the lobules were more likely to be actively dividing. Cells from ducts and lobules collected under the microscope were functionally characterized by colony formation on tissue culture plastic, mammosphere formation in suspension culture, and morphogenesis in laminin-rich extracellular matrix gels. Staining for the lineage markers keratins K14 and K19 further revealed multipotent cells in the stem cell zone and three lineage restricted cell types outside this zone. Multiparameter cell sorting and functional characterization with reference to anatomical sites in situ confirmed this pattern. The proposal that the four cell types are indeed constituents of an as of yet undescribed stem cell hierarchy was assessed in long-term cultures in which senescence was bypassed. These findings identify an adult human breast ductal stem cell activity and its earliest descendants. PMID- 17420293 TI - Grd19/Snx3p functions as a cargo-specific adapter for retromer-dependent endocytic recycling. AB - Amajor function of the endocytic system is the sorting of cargo to various organelles. Endocytic sorting of the yeast reductive iron transporter, which is composed of the Fet3 and Ftr1 proteins, is regulated by available iron. When iron is provided to iron-starved cells, Fet3p-Ftr1p is targeted to the lysosome-like vacuole and degraded. In contrast, when iron is not available, Fet3p-Ftr1p is maintained on the plasma membrane via an endocytic recycling pathway requiring the sorting nexin Grd19/Snx3p, the pentameric retromer complex, and the Ypt6p Golgi Rab GTPase module. A recycling signal in Ftr1p was identified and found to bind directly to Grd19/Snx3p. Retromer and Grd19/Snx3p partially colocalize to tubular endosomes, where they are physically associated. After export from the endosome, Fet3p-Ftr1p transits through the Golgi apparatus for resecretion. Thus, Grd19/Snx3p, functions as a cargo-specific adapter for the retromer complex, establishing a precedent for a mechanism by which sorting nexins expand the repertoire of retromer-dependent cargos. PMID- 17420294 TI - Adherens junction remodeling by the Notch pathway in Drosophila melanogaster oogenesis. AB - Identifying genes involved in the control of adherens junction (AJ) remodeling is essential to understanding epithelial morphogenesis. During follicular epithelium development in Drosophila melanogaster, the main body follicular cells (MBFCs) are displaced toward the oocyte and become columnar. Concomitantly, the stretched cells (StCs) become squamous and flatten around the nurse cells. By monitoring the expression of epithelial cadherin and Armadillo, I have discovered that the rate of AJ disassembly between the StCs is affected in follicles with somatic clones mutant for fringe or Delta and Serrate. This results in abnormal StC flattening and delayed MBFC displacement. Additionally, accumulation of the myosin II heavy chain Zipper is delayed at the AJs that require disassembly. Together, my results demonstrate that the Notch pathway controls AJ remodeling between the StCs and that this role is crucial for the timing of MBFC displacement and StC flattening. This provides new evidence that Notch, besides playing a key role in cell differentiation, also controls cell morphogenesis. PMID- 17420295 TI - Deficiency in the LIM-only protein Fhl2 impairs skin wound healing. AB - After skin wounding, the repair process is initiated by the release of growth factors, cytokines, and bioactive lipids from injured vessels and coagulated platelets. These signal molecules induce synthesis and deposition of a provisional extracellular matrix, as well as fibroblast invasion into and contraction of the wounded area. We previously showed that sphingosine-1 phosphate (S1P) triggers a signal transduction cascade mediating nuclear translocation of the LIM-only protein Fhl2 in response to activation of the RhoA GTPase (Muller, J.M., U. Isele, E. Metzger, A. Rempel, M. Moser, A. Pscherer, T. Breyer, C. Holubarsch, R. Buettner, and R. Schule. 2000. EMBO J. 19:359-369; Muller, J.M., E. Metzger, H. Greschik, A.K. Bosserhoff, L. Mercep, R. Buettner, and R. Schule. 2002. EMBO J. 21:736-748.). We demonstrate impaired cutaneous wound healing in Fhl2-deficient mice rescued by transgenic expression of Fhl2. Furthermore, collagen contraction and cell migration are severely impaired in Fhl2-deficient cells. Consequently, we show that the expression of alpha-smooth muscle actin, which is regulated by Fhl2, is reduced and delayed in wounds of Fhl2-deficient mice and that the expression of p130Cas, which is essential for cell migration, is reduced in Fhl2-deficient cells. In summary, our data demonstrate a function of Fhl2 as a lipid-triggered signaling molecule in mesenchymal cells regulating their migration and contraction during cutaneous wound healing. PMID- 17420296 TI - Tumor priming enhances delivery and efficacy of nanomedicines. AB - We have shown that high epithelial cell density is a major barrier to the distribution of protein-bound drugs in solid tumors, and tumor priming (expansion of interstitial space using an apoptosis-inducing pretreatment) can promote drug delivery. This study evaluated the optimal conditions of paclitaxel tumor priming (time window, particle size) and its effects on the delivery and efficacy of nanomedicines. Paclitaxel tumor priming was applied to mice bearing human xenograft tumors. The kinetics of paclitaxel-induced apoptosis was evaluated to identify the time window of tumor priming. The effects of tumor priming on the tumor delivery and interstitial dispersion of fluorescence-labeled nanoparticles of various sizes, the perfusion of tumor and normal tissues, the delivery of doxorubicin HCl liposomes to tumor and host tissues, and the antitumor activity and host toxicity were studied. Tumor priming by a single i.v. injection of paclitaxel induced apoptosis, expanded the interstitial space, vessel diameter and blood-perfused area, and promoted the delivery and interstitial dispersion of nanoparticles (100- and 200-nm diameter, administered 48 h after paclitaxel) in a tumor-selective manner. Tumor priming also enhanced the tumor delivery and antitumor activity of doxorubicin HCl liposomes (85 nm) without affecting the delivery to noncancerous host tissues or enhancing host toxicity. Tumor priming represents a potentially useful means to promote tumor-selective delivery and efficacy of nanomedicines. The current study will have significant impact on enhancing delivery and efficacy of nanomedicines and dosing regimen optimization of combination chemotherapy in the clinical setting. PMID- 17420297 TI - Biomarker optimization to track the antithrombotic and hemostatic effects of clopidogrel in rats. AB - We determined the dose response of the ADP antagonist clopidogrel (0.3-50 mg/kg p.o.) in rat models of thrombosis and provoked bleeding and correlated these activities to ex vivo platelet activation. Carotid artery thrombosis was induced by FeCl(2). Bleeding time was measured by mesenteric vessel puncture and renal cortex or cuticle incision. Platelet biomarkers included standard ADP-induced aggregation, P2Y(12) receptor occupancy, and phosphorylation of vasodilator stimulated phosphoprotein. Clopidogrel decreased thrombus weight up to 78%, caused maximal prolongation of cuticle and mesenteric bleeding, but had little effect on renal bleeds. Due to the steep mesenteric dose response, further comparisons concentrated on cuticle bleeding. The half-maximal inhibitory dose (ED(50)) for thrombus reduction was 2.4 +/- 0.4 mg/kg, with 10 mg/kg providing optimal blood flow preservation and thrombus reduction. The ED(50) for bleeding was 10.5 +/- 3.4 mg/kg. Increased bleeding was intermediate (3-fold) at 10 mg/kg and maximal (6-fold) at 30 mg/kg. All biomarkers were affected, but with differing sensitivity. ED(50)s for peak platelet aggregation to 10 microM ADP (11.9 +/- 0.4 mg/kg) and the vasodilator-stimulated phosphoprotein index (16.4 +/ 1.3 mg/kg) approximated the higher ED(50) for bleeding. ED(50)s for ligand binding (3.0 +/- 0.3 mg/kg) and late aggregation (5.1 +/- 0.4 mg/kg) better matched the lower ED(50) for antithrombotic activity. Aspirin exerted lesser effects on bleeding (42-70% increase in all models) and thrombosis (24% inhibition). In summary, antithrombotic doses of clopidogrel have limited effects on bleeding and standard measures of platelet aggregation. Other biomarkers may be more sensitive for tracking antithrombotic efficacy. PMID- 17420298 TI - Online teaching resources: where in the WWW are they? PMID- 17420299 TI - A dominant role of cardiac molecular motors in the intrinsic regulation of ventricular ejection and relaxation. AB - Molecular motors housed in myosins of the thick filament react with thin-filament actins and promote force and shortening in the sarcomeres. However, other actions of these motors sustain sarcomeric activation by cooperative feedback mechanisms in which the actin-myosin interaction promotes thin-filament activation. Mechanical feedback also affects the actin-myosin interaction. We discuss current concepts of how these relatively under-appreciated actions of molecular motors are responsible for modulation of the ejection time and isovolumic relaxation in the beating heart. PMID- 17420300 TI - Gene therapy in the treatment of heart failure. AB - Heart failure is a major cause of morbidity and mortality in contemporary societies. Although progress in conventional treatment modalities is making steady and incremental gains to reduce this disease burden, there remains a need to explore new and potentially therapeutic approaches. Gene therapy, for example, was initially envisioned as a treatment strategy for inherited monogenic disorders. It is now apparent that gene therapy has broader potential that also includes acquired polygenic diseases, such as heart failure. Advances in the understanding of the molecular basis of conditions such as these, together with the evolution of increasingly efficient gene transfer technology, has placed congestive heart failure within reach of gene-based therapy. PMID- 17420301 TI - Transport and peripheral bioactivities of nitrogen oxides carried by red blood cell hemoglobin: role in oxygen delivery. AB - The biology of NO (nitric oxide) is poorly explained by the activity of the free radical NO ((.)NO) itself. Although (.)NO acts in an autocrine and paracrine manner, it is also in chemical equilibrium with other NO species that constitute stable stores of NO bioactivity. Among these species, S-nitrosylated hemoglobin (S-nitrosohemoglobin; SNO-Hb) is an evolved transducer of NO bioactivity that acts in a responsive and exquisitely regulated manner to control cardiopulmonary and vascular homeostasis. In SNO-Hb, O(2) sensing is dynamically coupled to formation and release of vasodilating SNOs, endowing the red blood cell (RBC) with the capacity to regulate its own principal function, O(2) delivery, via regulation of blood flow. Analogous, physiological actions of RBC SNO-Hb also contribute to central nervous responses to blood hypoxia, the uptake of O(2) from the lung to blood, and baroreceptor-mediated control of the systemic flow of blood. Dysregulation of the formation, export, or actions of RBC-derived SNOs has been implicated in human diseases including sepsis, sickle cell anemia, pulmonary arterial hypertension, and diabetes mellitus. Delivery of SNOs by the RBC can be harnessed for therapeutic gain, and early results support the logic of this approach in the treatment of diseases as varied as cancer and neonatal pulmonary hypertension. PMID- 17420302 TI - The ins and outs of secretion from pancreatic beta-cells: control of single vesicle exo- and endocytosis. AB - Exocytosis of insulin-containing secretory vesicles in pancreatic beta-cells is crucial to maintenance of plasma glucose levels. They fuse with the plasma membrane in a regulated manner to release their contents and are subsequently recaptured either intact or through conventional clathrin-mediated endocytosis. Here, we discuss these mechanisms in beta-cells at the single-vesicle level. PMID- 17420303 TI - Structure and function of ABC transporters. AB - ATP binding cassette transporters are ubiquitous integral membrane proteins that actively transport ligands across biological membranes, a process critical for most aspects of cell physiology. These proteins are important clinically and economically. Their dysfunction underlies a number of human genetic diseases, and the ability of some to pump cytotoxic molecules from cells confers resistance to antibiotics, herbicides, and chemotherapeutic drugs. Recent structure analyses interpreted in light of a large body of biochemistry has resulted in the ATP switch model for function in which the paired nucleotide binding domains switch between an ATP-dependent closed conformation and a nucleotide-free, open conformation to drive the translocation of ligand. PMID- 17420304 TI - Ca2+ signaling in the inner ear. AB - The inner ear contains delicate sensory receptors that have adapted to detect the minutest mechanical disturbances. Ca(2+) ions are implicated in all steps of the transduction process, as well as in its regulation by an impressive ensemble of finely tuned feedback control mechanisms. Recent studies have unveiled some of the key players, but things do not sound quite right yet. PMID- 17420305 TI - Direct and indirect cortico-motoneuronal pathways and control of hand/arm movements. AB - Recent studies from our group have demonstrated the existence of a disynaptic excitatory cortico-motoneuronal (CM) pathway in macaque monkeys via propriospinal neurons in the midcervical segments. Results from behavioral studies with lesion of the direct pathway suggest that the indirect CM pathway can mediate the command for dexterous finger movements. PMID- 17420306 TI - Risk factors for the development of pedal edema in patients using pramipexole. AB - OBJECTIVE: To determine risk factors for pedal edema among patients with Parkinson disease (PD) using pramipexole hydrochloride therapy. DESIGN: A retrospective medical record review. SETTING: Philadelphia Veterans Administration Parkinson's Disease Research, Education and Clinical Center (PADRECC). PATIENTS: All consecutive patients at the PADRECC receiving pramipexole from December 2002 to December 2004. MAIN OUTCOME MEASURES: Bivariable and multivariable logistic regression models were used to identify comorbid illnesses, demographic characteristics, other medications, and PD features associated with increased risk of pedal edema among individuals taking pramipexole. Estimation of time to development of pedal edema in individuals taking pramipexole was performed using Kaplan-Meier survival methods and multivariable Cox proportional hazards models. RESULTS: Two hundred thirty-seven PADRECC patients received pramipexole and met criteria for inclusion in the analysis. Of these, 38 (16%) developed pedal edema. Multivariable regression models identified idiopathic PD (odds ratio [OR], 4.80; 95% confidence interval [CI], 1.54-14.98; P = .007), history of coronary artery disease (OR, 3.35; 95% CI, 1.51-7.46; P = .003), and history of diabetes mellitus (OR, 3.12; 95% CI, 1.01-9.60; P = .05) as strong independent risk factors for development of edema. There was no relationship between dose of pramipexole and incidence and severity of pedal edema. The risk of development of pedal edema was 7.7% (95% CI, 4.5% 12.9%) in the first year after initiation of pramipexole therapy, with more rapid development of edema among those with a history of coronary artery disease. CONCLUSIONS: Pedal edema is a relatively common outcome in patients with PD receiving pramipexole. History of coronary artery disease increases the risk for developing edema. PMID- 17420308 TI - Mars on balance: twitching and dreaming. PMID- 17420309 TI - Space exploration, Mars, and the nervous system. AB - When human beings venture back to the moon and then on to Mars in the coming decade or so, we will be riding on the accumulated data and experience from approximately 50 years of manned space exploration. Virtually every organ system functions differently in the absence of gravity, and some of these changes are maladaptive. From a biologic perspective, long duration spaceflight beyond low Earth orbit presents many unique challenges. Astronauts traveling to Mars will live in the absence of gravity for more than 1 year en route and will have to transition between weightlessness and planetary gravitational forces at the beginning, middle, and end of the mission. We discuss some of what is known about the effects of spaceflight on nervous system function, with emphasis on the neuromuscular and vestibular systems because success of a Mars mission will depend on their proper functioning. PMID- 17420310 TI - Superficial siderosis: associations and therapeutic implications. AB - Superficial siderosis of the central nervous system results from hemosiderin deposition in the subpial layers of the brain and spinal cord. A clinical history of subarachnoid hemorrhage is often absent. Patients present with slowly progressive gait ataxia and sensorineural hearing impairment. A history of prior intradural surgery or trauma is common. With widespread use of magnetic resonance imaging, presymptomatic cases are being diagnosed and it is difficult to be certain about the true incidence of this disorder. Despite extensive investigations, the cause of bleeding is often not apparent. An intraspinal fluid filled collection is a common accompaniment and may be the likely bleeding source. An early diagnosis and prompt intervention directed at removal of the bleeding source may prevent progression. This review discusses the role of multimodality imaging in evaluation of superficial siderosis and the therapeutic implications of identified associations. PMID- 17420311 TI - The association between genetic variants in SORL1 and Alzheimer disease in an urban, multiethnic, community-based cohort. AB - OBJECTIVE: To investigate the association between Alzheimer disease (AD) and variant alleles in SORL1 using a series of single nucleotide polymorphisms (SNPs) in an urban, multiethnic, community-based population. DESIGN: We used a nested case-control analysis in a population-based, prospective study of aging and dementia in Medicare recipients, 65 years and older. SETTING: Northern Manhattan, NY. PARTICIPANTS: There were 296 patients with probable AD and 428 healthy, elderly controls. The participants were African American (34%), Caribbean Hispanic (51%), or non-Hispanic white (15%). MAIN OUTCOME MEASURES: We genotyped all 29 SNPs in SORL1 that were examined in the earlier report. We assessed allelic association with AD using standard case-control methods, which included apolipoprotein E genotype as a covariate. RESULTS: Several individual SNPs and SNP haplotypes were significantly associated with AD in this prospectively collected community-based cohort, confirming the previously reported positive association of SORL1 with AD. Single nucleotide polymorphism 12, near the 5' region, was associated with AD in African American and Hispanic individuals. Two SNPs in the 3' region were also associated with AD in African American (SNP 26) and non-Hispanic white (SNP 20) individuals. A single haplotype in the 3' region was associated with AD in Hispanic individuals. However, several different haplotypes were associated with AD in African American and white individuals, including the TTC haplotypes at SNPs 23 through 25 (P = .035), which was significantly associated with AD in the North European white individuals in our previous report. CONCLUSIONS: This study confirms the association between genetic variants in SORL1 and AD. While the associations observed in these data sets overlap with those previously reported, the finding of novel SNP and haplotype associations suggests that there may be extensive allelic heterogeneity in SORL1. Broad regions of the SORL1 gene will therefore need to be scrutinized for functional pathogenic variants. PMID- 17420312 TI - Slow progressive acceptance of intravenous thrombolysis for patients with stroke by rural primary care physicians. AB - BACKGROUND: In the rural United States, patients with stroke are usually first evaluated locally by a nonneurologist physician (NNP) before treatment is determined. OBJECTIVE: To determine the evolution of NNPs' familiarity and attitudes about using recombinant tissue plasminogen activator (rtPA) since this therapy has been approved. DESIGN: Cross-sectional design using 2 similar surveys mailed in 1997 and 2003 to all primary care, family, internal, and emergency medicine physicians in the state of Iowa (1582 and 1679 physicians, respectively). PARTICIPANTS: All NNPs (primary care, internal, and emergency medicine) practicing in the state of Iowa. MAIN OUTCOME MEASURES: Comparison of 1997 and 2003 aggregate responses to questions about familiarity and willingness to use rtPA to treat patients who have had an acute ischemic stroke. RESULTS: The willingness of NNPs to use rtPA to treat acute ischemic stroke increased from 18% to 32% between 1997 and 2003. The number of NNPs who were very familiar with the National Institutes of Health Stroke Scale increased from 1% to 13%. Compared with physicians in 1997, more physicians in 2003 knew that prolonged international normalized ratios (42% vs 61%) or excessively high blood pressures (61% vs 78%) were contraindications for the use of rtPA. Still, half of the respondents perceived that they were inadequately exposed to educational material about rtPA during these years. Most expressed preference for personal methods of delivery for future educational efforts. CONCLUSIONS: The familiarity and comfort among NNPs with the administration of rtPA is still relatively low in rural settings. The improvement observed between the years 1997 and 2003 is encouraging. The responses suggest that NNPs' acceptance of rtPA can be further improved with educational campaigns involving personal methods of delivery. PMID- 17420313 TI - Disease course and prognostic factors of progressive muscular atrophy. AB - OBJECTIVE: To investigate the natural history and prognostic factors in patients with nonhereditary, adult-onset progressive muscular atrophy. DESIGN: Inception cohort conducted for 18 months. Settings Three university hospitals in the Netherlands (referral centers for neuromuscular diseases). Patients Thirty-seven consecutive patients newly diagnosed (onset of weakness <4 years) with progressive muscular atrophy enrolled between 1998 and 2001. MAIN OUTCOME MEASURES: Disease progression was measured at 0, 3, 6, 9, 12, 15, and 18 months by the Medical Research Council sum score, number of affected limb regions, and the Amyotrophic Lateral Sclerosis Functional Rating Scale score. Multivariate linear regression analysis was used to identify predictors of poor outcome. Clinical features and classification of phenotype during follow-up were evaluated. Survival analysis was planned after data collection, performed 5 years after the end of the study. RESULTS: Significant decline of muscle strength (mean, 6.01 Medical Research Council sum score points [95% confidence interval [CI], 3.84-8.18]; P value <.001) and significant increase in the number of affected regions (mean, 0.53 affected region [95% CI, 0.42-0.65]; P value <.001) and functional impairment (mean, 1.85 Amyotrophic Lateral Sclerosis Functional Rating Scale score points [95% CI, 1.38-2.33]; P value <.001) were found. Vital capacity (VC) at baseline and decrease of VC during the first 6 months were significantly associated with outcome. Median survival duration after initial weakness was 56 months. CONCLUSIONS: This study shows that patients with progressive muscular atrophy have a relentlessly progressive disease course. Patients with a low VC at baseline and a sharp decline of VC during the first 6 months have an especially poor prognosis. PMID- 17420314 TI - Continuum of frontal lobe impairment in amyotrophic lateral sclerosis. AB - OBJECTIVE: To identify the nature and prevalence of cognitive and behavioral abnormalities in patients with amyotrophic lateral sclerosis (ALS). DESIGN: Survey of clinical characteristics. SETTING: Multidisciplinary clinic within a university medical center. Patients A volunteer sample of 30 new patients with ALS were recruited consecutively. Of those invited, 23 participants (20 with sporadic ALS and 3 with familial ALS) enrolled. Participants ranged in age from 27 to 80 years (mean age, 56.5 years); the education level ranged from 12 to 21 years (mean education level, 3.5 years of college); and 17 participants (74%) were male. MAIN OUTCOME MEASURES: Neuropsychological tests, neurobehavioral interviews, and structured magnetic resonance imaging. RESULTS: Patients were classified into subtypes of frontotemporal lobar degeneration (n = 5), suspected Alzheimer disease (n = 1), and subthreshold variants of cognitive impairment (n = 2), behavioral impairment (n = 4), and cognitively and behaviorally normal (n = 11). Five neuropsychological tests, 2 behavioral abnormalities, and right hemisphere gray matter reductions differentiated patients into normal and abnormal groups. CONCLUSIONS: In this sample, a sizable proportion of patients with ALS possess a range of behavioral and cognitive changes that lie on a spectrum of frontotemporal impairment. Right hemisphere atrophy may be a biomarker for cognitive impairment in patients with ALS. PMID- 17420315 TI - Clinical and psychometric distinction of frontotemporal and Alzheimer dementias. AB - BACKGROUND: A proportion of patients who meet the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Associations criteria for Alzheimer disease (AD) have frontotemporal lobar degeneration (FTLD) confirmed at autopsy, with or without concomitant AD. Thus, the clinical phenotypes of the 2 disorders may overlap. OBJECTIVE: To identify clinical and psychometric indicators that distinguish AD from FTLD at initial presentation. DESIGN: Longitudinal study of memory and aging. SETTING: Alzheimer's Disease Research Center, Washington University School of Medicine. PARTICIPANTS: Forty-eight clinically well-characterized cases of autopsy-confirmed FTLD (27 with psychometric testing results) were compared with 27 autopsy-confirmed AD cases. RESULTS: Behavioral abnormalities, particularly impulsivity (P<.001), disinhibition (P<.001), social withdrawal (P = .01), and progressive nonfluent aphasia, distinguished individuals with FTLD from those with AD. The individuals with FTLD performed better than those with AD on a visual test of episodic memory (P = .01), but worse on word fluency (P = .02) (performance correlated with aphasic features). Other cognitive and clinical features, including executive dysfunction and memory impairment, were comparable between the FTLD and AD groups. Concomitant histopathological AD was present in 11 of the 48 individuals with FTLD. CONCLUSIONS: Clinical and cognitive features of FTLD may overlap with AD, although behavioral and language difficulties distinguish those with FTLD. Memory loss in those with FTLD may in part reflect word-finding difficulties stemming from language dysfunction. Compounding the overlap of FTLD and AD clinical phenotypes is the presence of histopathological AD in almost one fourth of individuals with FTLD. PMID- 17420316 TI - Association of increased cortical soluble abeta42 levels with diffuse plaques after severe brain injury in humans. AB - BACKGROUND: Traumatic brain injury (TBI) is an environmental risk factor for developing Alzheimer disease. This may be due, in part, to changes associated with beta-amyloid (Abeta) plaque formation, which can occur within hours after injury, regardless of the patient's age. In addition to being precursors of toxic fibrils that deposit into plaques, soluble (nonfibrillar) Abeta peptides are posited to disrupt synaptic function and are associated with cognitive decline in Alzheimer disease. Changes in soluble Abeta levels and their relationship to Abeta plaque formation following TBI are unknown. OBJECTIVE: To quantify brain tissue levels of soluble Abeta peptides and their precursor protein in relation to Abeta plaque formation after TBI in humans. DESIGN: Surgically resected temporal cortex tissue from patients with severe TBI was processed for biochemical assays of soluble Abeta peptides with COOH-termini ending in amino acid 40 (Abeta(40)) or 42 (Abeta(42)) and Abeta precursor protein to compare patients with cortical Abeta plaques and those without. Patients Nineteen subjects admitted to the University of Pittsburgh Medical Center for treatment of severe closed head injury. RESULTS: Patients with severe TBI and cortical plaques had higher levels of soluble Abeta(1-42) but not Abeta(1-40); half of them were apolipoprotein E (APOE) epsilon4 allele carriers. The lowest Abeta levels were in 1 patient without plaques who was the only subject with an APOE epsilon2 allele. beta-Amyloid precursor protein levels were comparable in the 2 TBI groups. CONCLUSIONS: Selective increases in soluble Abeta(1-42) after TBI may predispose individuals with a brain injury to Alzheimer disease pathology. This may be influenced by the APOE genotype, and it may confer increased risk for developing Alzheimer disease later in life. PMID- 17420317 TI - Multiplex families with multiple system atrophy. AB - BACKGROUND: Multiple system atrophy (MSA) has been considered a sporadic disease, without patterns of inheritance. OBJECTIVE: To describe the clinical features of 4 multiplex families with MSA, including clinical genetic aspects. DESIGN: Clinical and genetic study. SETTING: Four departments of neurology in Japan. Patients Eight patients in 4 families with parkinsonism, cerebellar ataxia, and autonomic failure with age at onset ranging from 58 to 72 years. Two siblings in each family were affected with these conditions. MAIN OUTCOME MEASURES: Clinical evaluation was performed according to criteria by Gilman et al. Trinucleotide repeat expansion in the responsible genes for the spinocerebellar ataxia (SCA) series and for dentatorubral-pallidoluysian atrophy (DRPLA) was evaluated by polymerase chain reaction. Direct sequence analysis of coding regions in the alpha-synuclein gene was performed. RESULTS: Consanguineous marriage was observed in 1 of 4 families. Among 8 patients, 1 had definite MSA, 5 had probable MSA, and 2 had possible MSA. The most frequent phenotype was MSA with predominant parkinsonism, observed in 5 patients. Six patients showed pontine atrophy with cross sign or slitlike signal change at the posterolateral putaminal margin or both on brain magnetic resonance imaging. Possibilities of hereditary ataxias, including SCA1 (ataxin 1, ATXN1), SCA2 (ATXN2), Machado-Joseph disease/SCA3 (ATXN1), SCA6 (ATXN1), SCA7 (ATXN7), SCA12 (protein phosphatase 2, regulatory subunit B, beta isoform; PP2R2B), SCA17 (TATA box binding protein, TBP) and DRPLA (atrophin 1; ATN1), were excluded, and no mutations in the alpha-synuclein gene were found. CONCLUSIONS: Findings in these multiplex families suggest the presence of familial MSA with autosomal recessive inheritance and a genetic predisposition to MSA. Molecular genetic approaches focusing on familial MSA are expected to provide clues to the pathogenesis of MSA. PMID- 17420318 TI - Mutation of the linker region of the polymerase gamma-1 (POLG1) gene associated with progressive external ophthalmoplegia and Parkinsonism. AB - OBJECTIVE: To define the molecular basis of the autosomal dominant progressive external ophthalmoplegia and parkinsonism in a large family with a dominantly transmitted multiple mitochondrial DNA deletion disorder. DESIGN: Microsatellite analysis and screening of the progressive external ophthalmoplegia 1 (PEO1), adenine nucleotide translocator 1 (ANT1), and polymerase gamma-1 (POLG1) genes. RESULTS: We identified 3 novel heterozygous POLG1 substitutions in the same family. Autosomal dominant progressive external ophthalmoplegia segregated with 1532G>A in exon 8 and an intronic variant c.2070 + 158G>A in cis. The one patient with parkinsonism had an additional heterozygous substitution in exon 7 in trans (1389G>T). Both coding region mutations were predicted to alter conserved amino acids in the linker region of polymerase gamma. None of the substitutions were found in 192 ethnically matched control chromosomes, 108 patients with progressive external ophthalmoplegia, nor 140 cases of sporadic idiopathic Parkinson disease. CONCLUSION: Both autosomal dominant progressive external ophthalmoplegia and parkinsonism can because caused by mutations that directly affect the polymerase domain of polymerase gamma. PMID- 17420319 TI - Neurological, cardiological, and oculomotor progression in 104 patients with Friedreich ataxia during long-term follow-up. AB - BACKGROUND: Friedreich ataxia (FA) is the most frequent autosomal recessive cerebellar ataxia. Although the phenotype is well known, disease progression has not been evaluated in a prospective manner. OBJECTIVE: To perform a long-term prospective follow-up of neurological, cardiological, and oculomotor function in patients with FA (FA patients). DESIGN: In this open-labeled prospective survey, we examined 104 FA patients every 6 months during a median period of 5 years (range, 6 months to 7 years), with a systematic standardized protocol. Data are reported as mean +/- SD. SETTING: Neurological examinations were performed at the Federation of Neurology and the Department of Genetics of the Salpetriere Hospital, Paris, France. Cardiological follow-up was performed at the Department of Cardiology; oculomotor examinations were performed at the Institut National de la Sante et de la Recherche Medicale Unit 679, at the same hospital. Patients We studied 104 FA patients with a confirmed molecular diagnosis. None were receiving antioxidant therapy at baseline; 88 accepted treatment with the coenzyme Q(10) analogue idebenone (5 mg/kg per day). Sixteen preferred not to be treated. INTERVENTIONS: Neurological status was evaluated with the International Cooperative Ataxia Rating Scale (ICARS) and a quantitative writing test. Cardiological evaluations included echocardiography, electrocardiography, and Holter monitoring. Oculomotor function was evaluated by electro-oculography to determine the frequency of square wave jerks. RESULTS: The total ICARS score worsened during follow-up, whether or not the patients were treated with idebenone (1.93 +/- 0.25 and 4.43 +/- 1.56 points per year, respectively). The total ICARS score increased faster in patients with onset before age 15 years compared with the others (2.6 +/- 0.4 [n = 51] vs 1.1 +/- 0.3 [n = 37]; P = .05). The posture subscore increased faster in patients able to stand at baseline, who also had shorter disease durations than patients unable to stand (1.25 +/- 0.12 vs 0.47 +/- 0.22 point per year; P<.001). Neurological progression was underestimated, however, by the ICARS scores, which reached a plateau in patients with long disease durations. Oculomotor function slightly deteriorated (0.09 +/- 0.02 Hz per year; P<.001). Left ventricular mass index decreased (-4.1 +/- 1.5 g/m(2) per year; P = .008), as did ejection fraction (-1.32% +/- 0.29% per year; P<.001). CONCLUSIONS: The neurological condition of FA patients deteriorated slowly over time, even with idebenone treatment. Although cardiac hypertrophy decreased under treatment, cardiac function did not improve. The ICARS scale is not appropriate to evaluate the progression of FA in patients with long disease durations. Additional quantitative measures may improve the reliability of this scale. PMID- 17420320 TI - Relation of diabetes to mild cognitive impairment. AB - BACKGROUND: Type 2 diabetes mellitus is an important risk factor for Alzheimer disease and is more prevalent in elderly minority persons compared with non Hispanic white persons. OBJECTIVE: To determine whether diabetes is related to a higher risk of mild cognitive impairment (MCI), a transitional stage between normal cognition and Alzheimer disease, in a multiethnic cohort with a high prevalence of diabetes. DESIGN: Longitudinal cohort study. SETTING: Northern Manhattan in New York, NY. PARTICIPANTS: We studied persons without prevalent MCI or dementia at baseline and with at least 1 follow-up interval. Of 1772 participants with a complete neuropsychological evaluation, 339 (19.1%) were excluded because of prevalent dementia, 304 were excluded because of prevalent MCI (17.2%), and 211 were excluded because of loss to follow-up (11.9%), resulting in a final sample of 918 participants for longitudinal analyses. MAIN OUTCOME MEASURES: We related diabetes defined by self-report to incident all cause MCI, amnestic MCI, and nonamnestic MCI. We conducted multivariate analyses with proportional hazards regression adjusting for age, sex, years of education, ethnic group, apolipoprotein E (APOE) epsilon4 allele, hypertension, low-density lipoprotein level, current smoking, heart disease, and stroke. RESULTS: A total of 334 persons had incident MCI, 160 (47.9%) had amnestic MCI, and 174 (52.1%) had nonamnestic MCI. Diabetes was related to a significantly higher risk of all cause MCI and amnestic MCI after adjustment for all covariates. Diabetes was also related to a higher risk of nonamnestic MCI, but this association was appreciably attenuated after adjustment for socioeconomic variables and vascular risk factors. The risk of MCI attributable to diabetes was 8.8% for the whole sample and was higher for African American persons (8.4%) and Hispanic persons (11.0%) compared with non-Hispanic white persons (4.6%), reflecting the higher prevalence of diabetes in minority populations in the United States. CONCLUSION: Diabetes is related to a higher risk of amnestic MCI in a population with a high prevalence of this disorder. PMID- 17420321 TI - Smoking, caffeine, and nonsteroidal anti-inflammatory drugs in families with Parkinson disease. AB - OBJECTIVE: To assess associations between Parkinson disease (PD) and putatively protective factors-smoking, caffeine (coffee, tea, and soft drinks), and nonsteroidal anti-inflammatory drugs (aspirin, ibuprofen, and naproxen). DESIGN: Family-based case-control study. SETTING: Academic medical center clinic. PARTICIPANTS: A total of 356 case subjects and 317 family controls who self reported environmental exposures. MAIN OUTCOME MEASURES: Associations between PD and environmental measures (history, status, dosage, duration, and intensity) of smoking, coffee, caffeine, nonsteroidal anti-inflammatory drugs, and non-aspirin nonsteroidal anti-inflammatory drugs were examined using generalized estimating equations with an independent correlation matrix while controlling for age and sex. RESULTS: Individuals with PD were significantly less likely to report ever smoking (odds ratio = 0.56; 95% confidence interval, 0.41-0.78). Additional measures of smoking revealed significant inverse associations with PD (P<.05) and trends in odds ratios (P<.005). Increasing intensity of coffee drinking was inversely associated with PD (test for trend P = .05). Increasing dosage (trend P = .009) and intensity (trend P = .01) of total caffeine consumption were also inversely associated, with high dosage presenting a significant inverse association for PD (odds ratio = 0.58; 95% confidence interval, 0.34-0.99). There were no significant associations between nonsteroidal anti-inflammatory drugs and PD. CONCLUSIONS: Inverse associations of smoking and caffeine were corroborated using families with PD, thus emphasizing smoking and caffeine as important covariates to consider in genetic studies of PD. PMID- 17420322 TI - Absence of beta-amyloid deposits after immunization in Alzheimer disease with Lewy body dementia. AB - OBJECTIVE: To describe the neuropathological and biochemical findings of the brain examination of a patient enrolled in the AN-1792(QS-21) trial with an initial clinical diagnosis of Alzheimer disease (AD), in whom Lewy body variant was thereafter clinically diagnosed. DESIGN: A case report. SETTING: University memory clinic. Patient A 74-year-old woman with clinical features of probable AD. Intervention The patient received 2 injections of 225 mug of AN-1792 (beta amyloid [Abeta]) plus 50 mug of the adjuvant QS-21 at an interval of 4 weeks. The patient was an antibody responder with an IgG anti-AN-1792 antibody titer exceeding 10 000 and an IgM titer exceeding 3500. Maximum serum anti-Abeta titers were reached in 4.7 months. During the 3 following years, while the Mini-Mental State Examination score remained globally stable despite several confusional episodes, she developed clinical features of dementia with Lewy bodies. The patient died 34 months postimmunization. An autopsy was performed. MAIN OUTCOME MEASURES: Neuropathological and biochemical examination of the brain using standardized evaluation for tau, beta-amyloid, and synuclein deposits. RESULTS: Neither neuropathological nor biochemical examinations showed amyloid deposit in the brain of this immunized patient. For tau deposition, Braak stage was IV/VI, and the Western blot analysis score was 9c/10. The neuropathological semiquantitative score for alpha-synuclein aggregation was 4. There was no inflammation. These results were compared with those of an age-matched patient with AD and a control devoid of any neurological disease. CONCLUSION: In this Lewy body variant case, with globally stable functional and cognitive features, Abeta immunization resulted in a significant clearance of amyloid deposits, with remaining tau and synuclein pathological features in the brain. Patients with a Lewy body variant of AD should not be excluded from enrollment in Abeta immunization trials. PMID- 17420323 TI - Reduced penetrance in a Brazilian family with spinocerebellar ataxia type 10. AB - OBJECTIVE: To describe reduced penetrance associated with early onset in a Brazilian family with spinocerebellar ataxia type 10. DESIGN: Clinical examination and molecular analysis for the ATTCT repeat responsible for spinocerebellar ataxia type 10 in a patient and family members through 3 generations. SETTING: Ambulatory care. Patients A 28-year-old female Brazilian patient who presented with early-onset cerebellar ataxia and epilepsy, and her 9 asymptomatic relatives. Main Outcome Measure Genotype-phenotype correlation. RESULTS: Molecular testing on this patient showed an expansion of approximately 850 ATTCT repeats at the SCA10 locus. Similar SCA10 expansions of approximately 850 repeats were identified in 6 of 8 asymptomatic paternal relatives examined. CONCLUSION: The stably transmitted pentanucleotide expansion of approximately 850 repeats may represent a mutant SCA10 allele with reduced penetrance that may express an early-onset, severe phenotype. PMID- 17420324 TI - Novel prion protein conformation and glycotype in Creutzfeldt-Jakob disease. AB - OBJECTIVE: To describe a novel molecular and pathological phenotype of Creutzfeldt-Jakob disease. Patient A 69-year-old woman with behavioral and personality changes followed by rapidly evolving dementia. RESULTS: Postmortem examination of the brain showed intracellular prion protein deposition and axonal swellings filled with amyloid fibrils. Biochemical analysis of the pathological prion protein disclosed a previously unrecognized PrP(Sc) tertiary structure lacking diglycosylated species. Genetic analysis revealed a wild-type prion protein gene. The prion agent responsible for this atypical phenotype was successfully passaged to bank voles. CONCLUSION: To our knowledge, our results define a new human prion disorder characterized by intracellular accumulation of a novel type of pathological prion protein. PMID- 17420325 TI - Creutzfeldt-Jakob disease with focal electroencephalographic and magnetic resonance imaging findings. PMID- 17420326 TI - The molar tooth sign of Joubert syndrome. PMID- 17420327 TI - Whole-diet approach, Mediterranean diet, and Alzheimer disease. PMID- 17420328 TI - New therapies for ataxia-telangiectasia. PMID- 17420329 TI - Central retinal venous occlusion and cerebral venous thrombosis. PMID- 17420331 TI - Pulse wave velocity is inversely related to vertebral bone density in hemodialysis patients. AB - Abnormalities of bone mineral metabolism in patients with stage-5 chronic kidney disease may contribute to the high incidence of cardiovascular disease. Noninvasive imaging methods may help predict the simultaneous presence of vasculopathy and bone disease. Accordingly, we measured pulse wave velocity and bone mineral density (BMD), and T-scores (number of SDs below the BMD of a younger reference group) of the spine by both dual energy x-ray absorptiometry and quantitative computed tomography (QCT) in 110 maintenance hemodialysis patients. Older age, white race, diabetes mellitus, lower diastolic blood pressure, and lower albumin levels were associated with lower QCT-assessed T scores (each P<0.05). After age and multivariable adjustment, pulse wave velocity (PWV) increased as QCT BMD decreased (the prevalence of PWV >or=9 m/s was 32.4%, 61.8%, and 76.5% for participants in the highest to the lowest tertile of QCT assessed BMD; P<0.001). In contrast, there was no relationship between spine dual energy x-ray absorptiometry-BMD and PWV. In unadjusted models, thoracic spine QCT assessed T-scores correlated significantly, albeit weakly, with aorta calcification (r=0.22; P=0.01) but not with coronary calcification. The odds ratio of PWV >or=9 m/s for patients taking vitamin D(3) or its analogs was 0.51 (95% CI: 0.19 to 1.39). In conclusion, low spine BMD is associated with increased PWV in stage-5 chronic kidney disease, supporting the notion of a close interaction of vascular and bone disease in this patient group. QCT and not dual energy x-ray absorptiometry should be used to assess spine BMD in dialysis patients. PMID- 17420332 TI - Difference in neutrophil superoxide generation during pregnancy between preeclampsia and essential hypertension. AB - This study investigated whether production of reactive oxygen species by neutrophils differs between women with preeclampsia and those with essential hypertension. First, we assessed superoxide production by neutrophils during pregnancy and 4 weeks after delivery in 15 healthy pregnant women, 12 women with preeclampsia, and 7 pregnant women with essential hypertension. We then examined effects of serum from each subject on superoxide production by neutrophils obtained from healthy nonpregnant women. Neutrophil superoxide production was measured by cytochrome C reduction. N-formyl-methionyl-leucyl-phenylalanine stimulated superoxide production was significantly increased in neutrophils from women with preeclampsia and women with essential hypertension compared with normal pregnant women. Four weeks postpartum, the level of superoxide production was significantly decreased in women with preeclampsia but not in women with either normal pregnancy or essential hypertension. When neutrophils obtained from nonpregnant women were preincubated with predelivery sera from each group, sera from women with preeclampsia significantly enhanced superoxide production compared with sera from the other 2 groups. When postpartum serum was used, enhancement of neutrophil superoxide production by serum from women with preeclampsia was significantly decreased compared with that by predelivery serum. In conclusion, increased neutrophil superoxide production resolved after delivery in preeclampsia, whereas activation persisted postpartum in women with essential hypertension. The different transition of neutrophil superoxide production in response to pregnancy appears to be that preeclampsia is characterized by the presence of serum factors that enhance neutrophil superoxide production. Thus, in preeclampsia, serum factors bear a more essential role producing superoxide than a behavior of neutrophils. PMID- 17420333 TI - Neuronal responsiveness to central Na+ in 2 congenic strains of Dahl salt sensitive rats. AB - Dahl salt-sensitive rats show increased Na(+) entry into the brain on high salt intake and increased sympathetic and pressor responses to central Na(+). We examined C10QTL2 and C17QTL to test whether they contribute to these phenotypes. In Dahl salt-sensitive, Lewis, and C10S.L16, and C17S.L2 congenic rats on a high salt diet for 8 to 10 days, blood pressure and heart rate were higher in Dahl salt-sensitive versus others and in C10S.L16 and C17S.L2 versus Lewis rats. Cerebrospinal fluid [Na(+)] increased by approximately 5 mmol/L in Dahl salt sensitive, C10S.L16, and C17S.L2 compared with Lewis rats. In rats on a regular salt diet, 8-minute intracerebroventricular infusions of artificial cerebrospinal fluid with increasing [Na(+)] caused increases in blood pressure, heart rate, and renal sympathetic nerve activity, which were approximately 90% larger in Dahl salt-sensitive and C17S.L2 versus Lewis rats and only 35% to 45% larger in C10S.L16 versus Lewis rats. In another set of rats on regular salt, blood pressure and heart rate were recorded by telemetry before and during intracerebroventricular infusion of Na(+)-rich cerebrospinal fluid for 14 days. Na(+)-rich cerebrospinal fluid caused significantly larger increases in blood pressure and heart rate, larger responses to air stress and more impairment of baroreflex in Dahl salt-sensitive and C17S.L2 rats versus Lewis rats. In contrast, responses in C10S.L16 rats were similar to those in Lewis rats. These data suggest that, in Dahl salt-sensitive rats, genetic variants in C10QTL2 but not C17QTL contribute to increased neuronal responsiveness to cerebrospinal fluid [Na(+)]. However, neither of them contributes to the increase in cerebrospinal fluid [Na(+)] induced by high salt. PMID- 17420334 TI - Junctional adhesion molecule-1 is upregulated in spontaneously hypertensive rats: evidence for a prohypertensive role within the brain stem. AB - Junctional adhesion molecule-1 (JAM-1) forms part of the tight junction between adjacent endothelial cells. Using microarray technology, we showed previously that JAM-1 was differentially expressed in the brain stem of spontaneously hypertensive rats compared with normotensive Wistar-Kyoto (WKY) rats. In this study, we quantified the expression of JAM-1 in the brain stem of spontaneously hypertensive rats and WKY rats and established whether any differential expression was confined to this region of the brain or was ubiquitous throughout the central nervous system and, indeed, the whole body. Because the nucleus tractus solitarii plays a pivotal role in arterial pressure regulation, we assessed whether JAM-1 in this region affects the chronic regulation of arterial pressure. Real time RT-PCR revealed that JAM-1 mRNA was upregulated in multiple regions of the brain and all of the peripheral vascular beds studied. In the nucleus tractus solitarii, the level of JAM-1 mRNA was significantly higher in both young (3-week-old, prehypertensive) and adult male spontaneously hypertensive rats (15 to 18 weeks old) than that of age-matched WKY rats (fold differences; prehypertensives: 1.01+/-0.06 versus 1.59+/-0.13; n=10; P<0.01; adult: 1.08+/-0.14 versus 2.86+/-0.57; n=10; P<0.01). After adenoviral-mediated expression of JAM-1 in the nucleus tractus solitarii of adult WKY rats (15 weeks old; n=6), systolic pressure was increased from 120+/-4 to 132+/-4 mm Hg (P<0.01). Our data suggest that JAM-1 expression in the spontaneously hypertensive rat is upregulated throughout the body compared with the WKY rat and that this is not secondary to the hypertension. When JAM-1 is expressed in the nucleus tractus solitarii, it raises arterial pressure, suggesting a novel prohypertensive role for this protein within the brain stem. PMID- 17420335 TI - Overexpression of myofibrillogenesis regulator-1 aggravates cardiac hypertrophy induced by angiotensin II in mice. AB - Myofibrillogenesis regulator-1 (MR-1) augments cardiomyocytes hypertrophy induced by angiotensin II (Ang II) in vitro. However, its roles in cardiac hypertrophy in vivo remain unknown. Here, we investigate whether MR-1 can promote cardiac hypertrophy induced by Ang II in vivo and elucidate the molecular mechanisms of MR-1 on cardiac hypertrophy. We used a model of Ang II-induced cardiac hypertrophy by infusion of Ang II in female mice. In wild-type mice subjected to the Ang II infusion, cardiac hypertrophy developed after 2 weeks. In mice overexpressing human MR-1 (transgenic), however, cardiac hypertrophy was significantly greater than in wild-type mice as estimated by heart weight:body weight ratio, cardiomyocyte area, and echocardiographic measurements, as well as cardiac atrial natriuretic peptide and B-type natriuretic peptide mRNA and protein levels. Our further results showed that cardiac inflammation and fibrosis observed in wild-type Ang II mice were augmented in transgenic Ang II mice. Importantly, increased nuclear factor kappaB activation was significantly increased higher in transgenic mice compared with wild-type mice after 2 weeks of Ang II infusion. In vitro experiments also revealed that overexpression of MR-1 enhanced Ang II-induced nuclear factor kappaB activation, whereas downregulation of MR-1 blocked it in cardiac myocytes. In conclusion, our results suggest that MR-1 plays an aggravative role in the development of cardiac hypertrophy via activation of the nuclear factor kappaB signaling pathway. PMID- 17420336 TI - Predictive utility of pulse pressure and other blood pressure measures for cardiovascular outcomes. AB - Data are sparse regarding the actual predictive utility of pulse pressure and other blood pressure measures for cardiovascular events. We included all of the participants from the Chicago Heart Association Detection Project in Industry who were free of cardiovascular disease and not receiving antihypertensive treatment at baseline (1967-1973). Baseline blood pressure measures were assessed for predictive utility for fatal and nonfatal events over 33 years. Among 36 314 participants (mean age: 39+/-13 years; 43.4% women), there were 11 452 deaths: 745 were attributed to stroke, 2812 to coronary disease, and 599 to heart failure. Of the 16 393 participants who attained Medicare eligibility, 3050, 1367, and 2207 had >or=1 hospitalization for stroke, myocardial infarction, or heart failure, respectively. In univariate analyses, hazards ratios for stroke death per SD of pulse, systolic, and diastolic pressure, respectively, were 1.49, 1.75, and 1.71. Likelihood ratio chi(2) (134.3, 302.0, and 232.6, respectively), Bayes information criteria values (15 142, 14 974, and 15 044, respectively), and areas under receiver-operating characteristic curves (0.59, 0.64, and 0.63, respectively) all indicated better predictive utility for systolic and diastolic compared with pulse pressure. Results for coronary or heart failure death and stroke, myocardial infarction, or heart failure hospitalization were similar. Pulse pressure had weaker predictive utility at all ages but particularly for those <50 years. In this large cohort study, pulse pressure had predictive utility for cardiovascular events that was inferior to systolic or diastolic pressure. These findings support the approach of current guidelines in the use of systolic and diastolic blood pressure to assess risk and the need for treatment. PMID- 17420337 TI - Adiponectin level and left ventricular hypertrophy in Japanese men. AB - A recent study has demonstrated that adiponectin inhibited hypertrophic signaling in the myocardium of mice, implying that a decrease in the blood adiponectin level could cause cardiac muscle hypertrophy. We hypothesized that a relationship might exist between the serum adiponectin level and electrocardiographically diagnosed left ventricular hypertrophy (ECG-LVH), and we examined this hypothesis by epidemiological study of 2839 Japanese male workers who were not taking medications for hypertension. ECG-LVH was defined as meeting Sokolow-Lyon voltage criteria and/or Cornell voltage-duration product. The subjects were categorized by tertiles of serum adiponectin level, and a multivariate logistic regression analysis was conducted relating left ventricular hypertrophy to adiponectin tertiles adjusting for potential confounding factors. Prevalence of ECG-LVH in the studied sample was 16.7%. Adiponectin ranged from 1.0 to 5.0 microg/mL in the lowest category and from 7.4 to 30.6 microg/mL in the highest. Compared with subjects in the highest adiponectin category, those in the lowest one had a significantly higher prevalence of ECG-LVH independent of age, body mass index, and systolic blood pressure with an odds ratio of 1.50 and a 95% CI of 1.16 to 1.94. Further adjustment for high-density lipoprotein cholesterol, triglyceride, and insulin resistance did not change the association (odds ratio: 1.68; 95% CI: 1.28 to 2.21; P<0.001). Similar results were obtained when different criteria for ECG-LVH were used or when subjects were stratified by blood pressure or body mass index. Adiponectin concentration was inversely and independently associated with ECG-LVH in Japanese men. PMID- 17420338 TI - Mechanism(s) of systolic blood pressure reduction and drug therapy in hypertension. PMID- 17420339 TI - World Hypertension Day 2007. PMID- 17420340 TI - Reduced ventricular volumes and improved systolic function with cardiac resynchronization therapy: a randomized trial comparing simultaneous biventricular pacing, sequential biventricular pacing, and left ventricular pacing. AB - BACKGROUND: Cardiac resynchronization therapy has emerged as an important therapy for advanced systolic heart failure. Among available cardiac resynchronization therapy pacing modes that restore ventricular synchrony, it is uncertain whether simultaneous biventricular (BiV), sequential BiV, or left ventricular (LV) pacing is superior. The Device Evaluation of CONTAK RENEWAL 2 and EASYTRAK 2: Assessment of Safety and Effectiveness in Heart Failure (DECREASE-HF) trial is the first randomized trial comparing these 3 cardiac resynchronization therapy modalities. METHODS AND RESULTS: The DECREASE-HF Trial is a multicenter trial in which 306 patients with New York Heart Association class III or IV heart failure, an LV ejection fraction < or = 35%, and a QRS duration > or = 150 ms were randomized to simultaneous BiV, sequential BiV, or LV pacing. LV volumes and systolic and diastolic function were assessed with echocardiography at baseline, 3 months, and 6 months. All groups had a significant reduction in LV end-systolic and end diastolic dimensions (P<0.001). The simultaneous BiV pacing group had the greatest reduction in LV end-systolic dimension (P=0.007). Stroke volume (P<0.001) and LV ejection fraction (P<0.001) improved in all groups with no difference across groups. CONCLUSIONS: Compared with LV pacing, simultaneous BiV pacing was associated with a trend toward greater improvement in LV size. There is little difference between simultaneous BiV pacing and sequential BiV pacing as programmed in this trial. PMID- 17420341 TI - Randomized trial comparing same-day discharge with overnight hospital stay after percutaneous coronary intervention: results of the Elective PCI in Outpatient Study (EPOS). AB - BACKGROUND: Percutaneous coronary intervention (PCI) in a day-case setting might reduce logistic constraints on hospital resources, but data on safety are limited. We evaluated the safety and feasibility of same-day discharge after PCI. METHODS AND RESULTS: Eight hundred consecutive patients scheduled for elective PCI by femoral approach were randomized to same-day discharge or overnight hospital stay. Four hours after PCI, patients were triaged as suitable for early discharge or not. Suitable patients were discharged immediately or kept overnight, according to randomization. Patients with an indication for extended hospital stay were not discharged regardless of randomization. Primary end points were death, myocardial infarction, coronary artery bypass graft surgery, repeat PCI, or puncture-related complications occurring within 24 hours after PCI. A total of 403 patients were assigned to same-day discharge, of whom 77 (19%) were identified for extended observation; 397 patients were assigned to overnight stay, of whom 85 (21%) were identified for extended observation. Among all patients, the composite primary end point occurred in 9 (2.2%) same-day discharge patients and in 17 (4.2%) overnight stay patients (risk difference, -0.020; 95% CI, -0.045 to -0.004; P for noninferiority <0.0001). Among patients deemed suitable for early discharge, the composite end point occurred in 1 of 326 (0.3%) same-day discharge patients and 2 of 312 (0.6%) overnight-stay patients (risk difference, -0.003; 95% CI, -0.014 to 0.007; P for noninferiority <0.0001). The last 3 events were related to puncture site. CONCLUSIONS: Same-day discharge after elective PCI is feasible and safe in the majority (80%) of patients selected for day-case PCI. Same-day discharge does not lead to additional complications compared with overnight stay. PMID- 17420342 TI - Compartmentalization of cardiac beta-adrenergic inotropy modulation by phosphodiesterase type 5. AB - BACKGROUND: Recent cell-based studies have found that cGMP synthesis and hydrolysis by phosphodiesterase (PDE) appear compartmentalized, with nitric oxide synthase-derived and/or PDE type 5 (PDE-5)-hydrolyzable cGMP undetected at the sarcolemmal membrane in contrast to cGMP stimulated by natriuretic peptide. In the present study, we determine the functional significance of such compartments with a comparison of beta-adrenergic modulation by PDE-5 inhibition to that of natriuretic peptide stimulation in both cardiomyocytes and intact hearts. The potential role of differential cGMP and protein kinase G stimulation by these 2 modulators was also studied. METHODS AND RESULTS: Intact C57/BL6 mouse hearts were studied with pressure-volume analysis, and adult isolated myocytes were studied with fluorescence microscopy. PDE-5 inhibition with 0.1 to 1 micromol/L sildenafil (SIL) suppressed isoproterenol (ISO)-stimulated contractility, whereas 10 micromol/L atrial natriuretic peptide (ANP) had no effect. ISO suppression by SIL was prevented in cells pretreated with a protein kinase G inhibitor. Surprisingly, myocardial cGMP changed little with SIL+ISO yet rose nearly 5-fold with ANP, whereas protein kinase G activation (vasodilator-stimulated protein phosphorylation; ELISA assay) displayed the opposite: increased with SIL+ISO but unaltered by ANP+ISO. PDE-5 and ANP compartments were functionally separated, as inhibition of nitric oxide synthase by N(w)-nitro-L-arginine methyl ester eliminated antiadrenergic effects of SIL, yet this was not restorable by co stimulation with ANP. CONCLUSIONS: Regulation of cardiac beta-adrenergic response by cGMP is specifically linked to a nitric oxide-synthesis/PDE-5-hydrolyzed pool signaling via protein kinase G. Natriuretic peptide stimulation achieves greater detectable increases in cGMP but not protein kinase G activity and does not modulate beta-adrenergic response. Such disparities likely contribute to differential cardiac regulation by drugs that modulate cGMP synthesis and hydrolysis. PMID- 17420343 TI - Defining obesity cut points in a multiethnic population. AB - BACKGROUND: Body mass index (BMI) is widely used to assess risk for cardiovascular disease and type 2 diabetes. Cut points for the classification of obesity (BMI >30 kg/m2) have been developed and validated among people of European descent. It is unknown whether these cut points are appropriate for non European populations. We assessed the metabolic risk associated with BMI among South Asians, Chinese, Aboriginals, and Europeans. METHODS AND RESULTS: We randomly sampled 1078 subjects from 4 ethnic groups (289 South Asians, 281 Chinese, 207 Aboriginals, and 301 Europeans) from 4 regions in Canada. Principal components factor analysis was used to derive underlying latent or "hidden" factors associated with 14 clinical and biochemical cardiometabolic markers. Ethnic-specific BMI cut points were derived for 3 cardiometabolic factors. Three primary latent factors emerged that accounted for 56% of the variation in markers of glucose metabolism, lipid metabolism, and blood pressure. For a given BMI, elevated levels of glucose- and lipid-related factors were more likely to be present in South Asians, Chinese, and Aboriginals compared with Europeans, and elevated levels of the blood pressure-related factor were more likely to be present among Chinese compared with Europeans. The cut point to define obesity, as defined by distribution of glucose and lipid factors, is lower by approximately 6 kg/m2 among non-European groups compared with Europeans. CONCLUSIONS: Revisions may be warranted for BMI cut points to define obesity among South Asians, Chinese, and Aboriginals. Using these revised cut points would greatly increase the estimated burden of obesity-related metabolic disorders among non-European populations. PMID- 17420344 TI - C-terminal provasopressin (copeptin) as a novel and prognostic marker in acute myocardial infarction: Leicester Acute Myocardial Infarction Peptide (LAMP) study. AB - BACKGROUND: The role of the vasopressin system after acute myocardial infarction is unclear. Copeptin, the C-terminal part of the vasopressin prohormone, is secreted stoichiometrically with vasopressin. We compared the prognostic value of copeptin and an established marker, N-terminal pro-B-type natriuretic peptide (NTproBNP), after acute myocardial infarction. METHODS AND RESULTS: In this prospective single-hospital study, we recruited 980 consecutive post-acute myocardial infarction patients (718 men, median [range] age 66 [24 to 95] years), with follow-up over 342 (range 0 to 764) days. Plasma copeptin was highest on admission (n=132, P<0.001, day 1 versus days 2 to 5) and reached a plateau at days 3 to 5. In the 980 patients, copeptin (measured at days 3 to 5) was elevated in patients who died (n=101) or were readmitted with heart failure (n=49) compared with survivors (median [range] 18.5 [0.6 to 441.0] versus 6.5 [0.3 to 267.0] pmol/L, P<0.0005). With logistic regression analysis, copeptin (odds ratio, 4.14, P<0.0005) and NTproBNP (odds ratio, 2.26, P<0.003) were significant independent predictors of death or heart failure at 60 days. The area under the receiver operating characteristic curves for copeptin (0.75) and NTproBNP (0.76) were similar. The logistic model with both markers yielded a larger area under the curve (0.84) than for NTproBNP (P<0.013) or copeptin (P<0.003) alone, respectively. Cox modeling predicted death or heart failure with both biomarkers (log copeptin [hazard ratio, 2.33], log NTproBNP [hazard ratio, 2.70]). In patients stratified by NTproBNP (above the median of approximately 900 pmol/L), copeptin above the median (approximately 7 pmol/L) was associated with poorer outcome (P<0.0005). Findings were similar for death and heart failure as individual end points. CONCLUSIONS: The vasopressin system is activated after acute myocardial infarction. Copeptin may predict adverse outcome, especially in those with an elevated NTproBNP (more than approximately 900 pmol/L). PMID- 17420345 TI - Global improvement of vascular function and redox state with low-dose folic acid: implications for folate therapy in patients with coronary artery disease. AB - BACKGROUND: Although dietary folate fortification lowers plasma homocysteine and may reduce cardiovascular risk, high-dose folic acid therapy appears to not alter clinical outcome. Folic acid and its principal circulating metabolite, 5 methyltetrahydrofolate, improve vascular function, but mechanisms relating folate dose to vascular function remain unclear. We compared the effects of folic acid on human vessels using pharmacological high-dose versus low-dose treatment, equivalent to dietary folate fortification. METHODS AND RESULTS: Fifty-six non folate-fortified patients with coronary artery disease were randomized to receive low-dose (400 microg/d) or high-dose (5 mg/d) folic acid or placebo for 7 weeks before coronary artery bypass grafting. Vascular function was quantified by magnetic resonance imaging before and after treatment. Vascular superoxide and nitric oxide bioavailability were determined in segments of saphenous vein and internal mammary artery. Low-dose folic acid increased nitric oxide-mediated endothelium-dependent vasomotor responses, reduced vascular superoxide production, and improved enzymatic coupling of endothelial nitric oxide synthase through availability of the cofactor tetrahydrobiopterin. No further improvement in these parameters occurred with high-dose compared with low-dose treatment. Whereas plasma 5-methyltetrahydrofolate increased proportionately with treatment dose of folic acid, vascular tissue 5-methyltetrahydrofolate showed no further increment with high-dose compared with low-dose folic acid. CONCLUSIONS: Low-dose folic acid treatment, comparable to daily intake and dietary fortification, improves vascular function through effects on endothelial nitric oxide synthase and vascular oxidative stress. High-dose folic acid treatment provides no additional benefit. These direct vascular effects are related to vascular tissue levels of 5-methyltetrahydrofolate rather than plasma levels. High-dose folic acid treatment likely confers no further benefit in subjects already receiving folate supplementation. PMID- 17420346 TI - Identifying heart failure patients at high risk for near-term cardiovascular events with serial health status assessments. AB - BACKGROUND: Identification of heart failure outpatients at increased risk for clinical deterioration remains a critical challenge, with few tools currently available to assist clinicians. We tested whether serial health status assessments with the Kansas City Cardiomyopathy Questionnaire (KCCQ) can identify patients at increased risk for mortality and hospitalization. METHODS AND RESULTS: We evaluated 1358 patients with heart failure after an acute myocardial infarction in the Eplerenone's Neurohormonal Efficacy and Survival Study, a multicenter randomized trial that included serial KCCQ assessments. Cox proportional-hazards models were used to examine whether changes in KCCQ scores during successive outpatient visits were independently associated with all-cause mortality and cardiovascular mortality or hospitalization. Change in KCCQ (deltaKCCQ) was linearly associated with all-cause mortality (hazard ratio [HR], for each 5-point decrease in deltaKCCQ, 1.11; 95% CI, 1.04 to 1.19) and the combined outcome of cardiovascular mortality or hospitalization (HR for each 5 point decrease in deltaKCCQ, 1.12; 95% CI 1.07 to 1.18). In Kaplan-Meier survival analysis, all-cause mortality among patients with deltaKCCQ of < or = -10, > -10 to < 10, and > 10 points was 26%, 16%, and 13%, respectively (P=0.008). After multivariable adjustment, the linear relationship between deltaKCCQ and both all cause mortality and combined cardiovascular death and hospitalization persisted (HR, 1.09; 95% CI, 1.00 to 1.18; and HR, 1.11; 95% CI, 1.05 to 1.17 for each 5 point decrease in deltaKCCQ, respectively). CONCLUSIONS: In heart failure outpatients, serial health status assessments with the KCCQ can identify high risk patients and may prove useful in directing the frequency of follow-up and the intensity of treatment. PMID- 17420347 TI - Instability in the diagnosis of metabolic syndrome in adolescents. AB - BACKGROUND: Factor analyses suggest that the structure underlying metabolic syndrome is similar in adolescents and adults. However, adolescence is a period of intense physiological change, and therefore stability of the underlying metabolic structure and clinical categorization based on metabolic risk is uncertain. METHODS AND RESULTS: We analyzed data from 1098 participants in the Princeton School District Study, a school-based study begun in 2001-2002, who were followed up for 3 years. We performed factor analyses of 8 metabolic risks at baseline and follow-up to assess stability of factor patterns and clinical categorization of metabolic syndrome. Metabolic syndrome was defined using the current American Heart Association/National Heart, Lung, and Blood Institute definition for adults (AHA), a modified AHA definition used in prior pediatric metabolic syndrome studies (pediatric AHA), and the International Diabetes Federation (IDF) guidelines. We found that factor structures were essentially identical at both time points. However, clinical categorization was not stable. Approximately half of adolescents with baseline metabolic syndrome lost the diagnosis at follow-up regardless of the definitions used: pediatric AHA=56% (95% confidence interval [CI], 42% to 69%), AHA=49% (95% CI, 32% to 66%), IDF=53% (95% CI, 38% to 68%). In addition to loss of the diagnosis, new cases were identified. Cumulative incidence rates were as follows: pediatric AHA=3.8% (95% CI, 2.8% to 5.2%); AHA=4.4% (95% CI, 3.3% to 5.9%); IDF=5.2% (95% CI, 4.0% to 6.8%). CONCLUSIONS: During adolescence, metabolic risk factor clustering is consistent. However, marked instability exists in the categorical diagnosis of metabolic syndrome. This instability, which includes both gain and loss of the diagnosis, suggests that the syndrome has reduced clinical utility in adolescence and that metabolic syndrome-specific pharmacotherapy for youth may be premature. PMID- 17420348 TI - Adherence to statin therapy under drug cost sharing in patients with and without acute myocardial infarction: a population-based natural experiment. AB - BACKGROUND: As medication spending grows, Medicare Part D will need to adapt its coverage policies according to emerging evidence from a variety of insurance policies. We sought to evaluate the consequences of copayment and coinsurance policies on the initiation of statin therapy after acute myocardial infarction and adherence to therapy in statin initiators using a natural experiment of all British Columbia residents aged 66 years and older. METHODS AND RESULTS: Three consecutive cohorts that included all patients who began statin therapy during full drug coverage (2001), coverage with a $10 or $25 copay (2002), and coverage with a 25% coinsurance benefit (2003-2004) were followed up with linked healthcare utilization data (n=51,561). Follow-up of cohorts was 9 months after each policy change. Adherence to statin therapy was defined as > or = 80% of days covered. Relative to full-coverage policies, adherence to new statin therapy was significantly reduced, from 55.8% to 50.5%, under a fixed copayment policy (-5.4% points; 95% CI, -6.4% to -4.4%) and the subsequent coinsurance policy (-5.4% points; 95% CI, -6.3% to -4.4%). An uninterrupted increase in the proportion of patients initiating statin therapy after an acute myocardial infarction (1.7% points per quarter) was observed over the study period, similar to a Pennsylvania control population with full coverage. Sudden changes to full out-of-pocket spending, similar to Medicare's Part D "doughnut hole," almost doubled the risk of stopping statins (adjusted odds ratio, 1.94, 95% CI, 1.82 to 2.08). CONCLUSIONS: Fixed patient copayment and coinsurance policies have negative effects on adherence to statin lipid-lowering drug therapy but not on their initiation after myocardial infarction. PMID- 17420349 TI - Lack of the antioxidant enzyme glutathione peroxidase-1 accelerates atherosclerosis in diabetic apolipoprotein E-deficient mice. AB - BACKGROUND: Recent clinical studies have suggested a major protective role for the antioxidant enzyme glutathione peroxidase-1 (GPx1) in diabetes-associated atherosclerosis. We induced diabetes in mice deficient for both GPx1 and apolipoprotein E (ApoE) to determine whether this is merely an association or whether GPx1 has a direct effect on diabetes-associated atherosclerosis. METHODS AND RESULTS: ApoE-deficient (ApoE-/-) and ApoE/GPx1 double-knockout (ApoE-/- GPx1 /-) mice were made diabetic with streptozotocin and aortic lesion formation, and atherogenic pathways were assessed after 10 and 20 weeks of diabetes. Aortic proinflammatory and profibrotic markers were determined by both quantitative reverse-transcription polymerase chain reaction analysis after 10 weeks of diabetes and immunohistochemical analysis after 10 and 20 weeks of diabetes. Sham injected nondiabetic counterparts served as controls. Atherosclerotic lesions within the aortic sinus region, as well as arch, thoracic, and abdominal lesions, were significantly increased in diabetic ApoE-/- GPx1-/- aortas compared with diabetic ApoE-/- aortas. This increase was accompanied by increased macrophages, alpha-smooth muscle actin, receptors for advanced glycation end products, and various proinflammatory (vascular cell adhesion molecule-1) and profibrotic (vascular endothelial growth factor and connective tissue growth factor) markers. Quantitative reverse-transcription polymerase chain reaction analysis showed increased expression of receptors for advanced glycation end products (RAGE), vascular cell adhesion molecule-1, vascular endothelial growth factor, and connective tissue growth factor. Nitrotyrosine levels were significantly increased in diabetic ApoE-/- GPx1-/- mouse aortas. These findings were observed despite upregulation of other antioxidants. CONCLUSIONS: Lack of functional GPx1 accelerates diabetes-associated atherosclerosis via upregulation of proinflammatory and profibrotic pathways in ApoE-/- mice. Our study provides evidence of a protective role for GPx1 and establishes GPx1 as an important antiatherogenic therapeutic target in patients with or at risk of diabetic macrovascular disease. PMID- 17420350 TI - Diagnostic thresholds for ambulatory blood pressure monitoring based on 10-year cardiovascular risk. AB - BACKGROUND: Current diagnostic thresholds for ambulatory blood pressure (ABP) mainly rely on statistical parameters derived from reference populations. We determined an outcome-driven reference frame for ABP measurement. METHODS AND RESULTS: We performed 24-hour ABP monitoring in 5682 participants (mean age 59.0 years; 43.3% women) enrolled in prospective population studies in Copenhagen, Denmark; Noorderkempen, Belgium; Ohasama, Japan; and Uppsala, Sweden. In multivariate analyses, we determined ABP thresholds, which yielded 10-year cardiovascular risks similar to those associated with optimal (120/80 mm Hg), normal (130/85 mm Hg), and high (140/90 mm Hg) blood pressure on office measurement. Over 9.7 years (median), 814 cardiovascular end points occurred, including 377 strokes and 435 cardiac events. Systolic/diastolic thresholds for optimal ABP were 116.8/74.2 mm Hg for 24 hours, 121.6/78.9 mm Hg for daytime, and 100.9/65.3 mm Hg for nighttime. Corresponding thresholds for normal ABP were 123.9/76.8, 129.9/82.6, and 110.2/68.1 mm Hg, respectively, and those for ambulatory hypertension were 131.0/79.4, 138.2/86.4, and 119.5/70.8 mm Hg. After rounding, approximate thresholds for optimal ABP amounted to 115/75 mm Hg for 24 hours, 120/80 mm Hg for daytime, and 100/65 mm Hg for nighttime. Rounded thresholds for normal ABP were 125/75, 130/85, and 110/70 mm Hg, respectively, and those for ambulatory hypertension were 130/80, 140/85, and 120/70 mm Hg. CONCLUSIONS: Population-based outcome-driven thresholds for optimal and normal ABP are lower than those currently proposed by hypertension guidelines. PMID- 17420351 TI - Lactadherin deficiency leads to apoptotic cell accumulation and accelerated atherosclerosis in mice. AB - BACKGROUND: Atherosclerosis is an immunoinflammatory disease; however, the key factors responsible for the maintenance of immune regulation in a proinflammatory milieu are poorly understood. METHODS AND RESULTS: Here, we show that milk fat globule-EGF factor 8 (Mfge8, also known as lactadherin) is expressed in normal and atherosclerotic human arteries and is involved in phagocytic clearance of apoptotic cells by peritoneal macrophages. Disruption of bone marrow-derived Mfge8 in a murine model of atherosclerosis leads to substantial accumulation of apoptotic debris both systemically and within the developing lipid lesions. The accumulation of apoptotic material is associated with a reduction in interleukin 10 in the spleen but an increase in interferon-gamma production in both the spleen and the atherosclerotic arteries. In addition, we report a dendritic cell dependent alteration of natural regulatory T-cell function in the absence of Mfge8. These events are associated with a marked acceleration of atherosclerosis. CONCLUSIONS: Lack of Mfge8 in bone marrow-derived cells enhances the accumulation of apoptotic cell corpses in atherosclerosis and alters the protective immune response, which leads to an acceleration of plaque development. PMID- 17420352 TI - Predictors of outcome in chronic thromboembolic pulmonary hypertension. AB - BACKGROUND: Chronic thromboembolic pulmonary hypertension (CTEPH) is characterized by intraluminal thrombus organization and fibrous obliteration of pulmonary arteries. Recently, associated medical conditions such as splenectomy, ventriculoatrial shunt for the treatment of hydrocephalus, permanent central intravenous lines, inflammatory bowel disease, and osteomyelitis were found to be associated with the development of CTEPH. The study aim was to define the impact of these novel risk factors on survival. METHODS AND RESULTS: Between January 1992 and December 2006, 181 patients diagnosed with CTEPH were tracked with the use of our center's customized computer database. A Cox regression model was used to examine relations between survival and associated medical conditions, age, sex, hemodynamic parameters, modified New York Heart Association functional class at diagnosis, CTEPH type, pulmonary endarterectomy, and anti-cardiolipin antibodies/lupus anticoagulant. During a median observation time of 22.1 (range, 0.03 to 152) months, the clinical end point of cardiovascular death or lung transplantation occurred in 48 cases (27%). Pulmonary endarterectomy (hazard ratio, 0.14; 95% CI, 0.05 to 0.41; P=0.0003), associated medical conditions (hazard ratio, 3.17; 95% CI, 1.70 to 5.92; P=0.0003), and pulmonary vascular resistance (hazard ratio, 1.02; 95% CI, 1.00 to 1.04; P=0.04) were predictors of survival. Thirty-day postoperative mortality (24% versus 9%) and the incidence of postoperative pulmonary hypertension (92% versus 20%) were substantially higher in patients with associated medical conditions. CONCLUSIONS: CTEPH-predisposing medical conditions, such as splenectomy, permanent central intravenous lines, and certain inflammatory disorders, predict poor survival in CTEPH. PMID- 17420353 TI - Optical visualization of cathepsin K activity in atherosclerosis with a novel, protease-activatable fluorescence sensor. AB - BACKGROUND: Cathepsin K (CatK), a potent elastinolytic and collagenolytic cysteine protease, likely participates in the evolution and destabilization of atherosclerotic plaques. To assess better the biology of CatK activity in vivo, we developed a novel near-infrared fluorescence (NIRF) probe for imaging of CatK and evaluated it in mouse and human atherosclerosis. METHODS AND RESULTS: The NIRF imaging agent consists of the CatK peptide substrate GHPGGPQGKC-NH2 linked to an activatable fluorogenic polymer. In vitro, CatK produced a 2- to 14-fold activation of the agent over other cysteine and matrix metalloproteinases (P<0.0001), as well as a >8-fold activation over a control imaging agent (P<0.001). Optical imaging of atheroma revealed >100% NIRF signal increases in apolipoprotein E-/- mice in vivo (n=13; P<0.05, CatK imaging agent versus control agent) and in human carotid endarterectomy specimens ex vivo (n=14; P<0.05). Fluorescence microscopy of plaque sections demonstrated that enzymatically active CatK (positive NIRF signal) localized primarily in the vicinity of CatK-positive macrophages. Augmented NIRF signal (reflecting CatK activity) colocalized with disrupted elastin fibers within the media underlying plaques. CONCLUSIONS: Use of this novel protease-activatable NIRF agent for optical imaging in vivo demonstrated preferential localization of enzymatically active CatK to macrophages, consistent with their known greater elastinolytic capabilities compared with smooth muscle cells. Augmented CatK proteolysis in atheromata further links CatK to vascular remodeling and plaque vulnerability. PMID- 17420354 TI - Mechanism underlying initiation of paroxysmal atrial flutter/atrial fibrillation by ectopic foci: a simulation study. AB - BACKGROUND: The mechanisms underlying paroxysmal atrial flutter/atrial fibrillation initiation by ectopic foci from various locations are unclear. METHODS AND RESULTS: We used parallel computational techniques to study an anatomically accurate 3-dimensional atrial structure incorporating a detailed ionic-current model of an atrial myocyte. At the single-cell level, upregulation of the L-type Ca2+ current I(Ca,L) steepened restitution curves of action potential duration and conduction velocity compared with the control. Spontaneous firings of ectopic foci, coupled with sinus activity, produced dynamic spatial dispersions of repolarization, including discordant alternans, which caused conduction block and reentry only for the elevated I(Ca,L) case. For each foci location, a vulnerable window for atrial flutter/atrial fibrillation induction was identified as a function of the coupling interval and focus cycle length. For ectopic foci in the pulmonary veins and left atrium, the site of conduction block and reentry gradually shifted, as a function of coupling interval, from the right atrium to the interatrial area and finally to the left atrium. The size of the vulnerable window was largest for pulmonary vein foci, becoming markedly smaller for right atrial foci, especially those near the sinoatrial node. CONCLUSIONS: These findings suggest that a mechanism of dynamically induced repolarization dispersion, especially discordant alternans, underlies the induction of atrial flutter/atrial fibrillation by atrial ectopic foci. The sites and likelihood of reentry induction varied according to ectopic focus location and timing, with the largest vulnerable window corresponding to the pulmonary vein region. PMID- 17420355 TI - Images in cardiovascular medicine. An unusual case of embolic stroke after permanent pacing. PMID- 17420356 TI - Images in cardiovascular medicine. Electron microscopic insights into the vascular biology of atherosclerosis: study of coronary endarterectomy specimens. PMID- 17420357 TI - Images in cardiovascular medicine. Rib perforation from a right ventricular pacemaker lead. PMID- 17420358 TI - Letter by Xia regarding article, "High-density lipoproteins and their constituent, sphingosine-1-phosphate, directly protect the heart against ischemia/reperfusion injury in vivo via the S1P3 lysophospholipid receptor". PMID- 17420359 TI - Spotlight: Jane Somerville, MD, FRCP, FESC. Interview by Robert Short. PMID- 17420360 TI - A direct comparison of intravascular ultrasound and quantitative coronary arteriography: implications for measures of atherosclerosis as clinical surrogates. PMID- 17420361 TI - Transgenic expression of A20 prevents cardiac cell death and myocardial dysfunction after myocardial infarction. PMID- 17420362 TI - New insights into pacemaker activity: promoting understanding of sick sinus syndrome. PMID- 17420363 TI - Adult congenital heart disease: right ventricular outflow tract lesions. PMID- 17420364 TI - Comparison of initial intraocular pressure response with topical beta-adrenergic antagonists and prostaglandin analogues in African American and white individuals in the Ocular Hypertension Treatment Study. AB - OBJECTIVE: To compare the intraocular pressure (IOP) responses of self-identified African American and white participants in the Ocular Hypertension Treatment Study to therapeutic trials of topical, nonselective beta-adrenergic antagonists or prostaglandin analogues. METHODS: Multivariate models that adjusted for baseline IOP and corneal thickness were used to estimate IOP response by race. Participants included 536 who were prescribed topical beta-adrenergic antagonists and 191 who were prescribed prostaglandin analogues, 25% of whom were African American. MAIN OUTCOME MEASURE: Intraocular pressure response in the ipsilateral eye after 4 to 6 weeks of a therapeutic trial. RESULTS: Intraocular pressure response to nonselective beta-adrenergic antagonists did not differ between African American and white participants. Intraocular pressure response to prostaglandin analogues was slightly greater in African American participants, but this difference was not statistically significant. With both classes of medication, greater IOP reduction was associated with higher baseline IOP and thinner central corneal measurement. CONCLUSIONS: We found no statistically significant differences in IOP response to topical, nonselective beta-adrenergic antagonists or prostaglandin analogues between self-identified African American and white individuals. Application to Clinical Practice Studies of IOP response to medication should statistically adjust for baseline IOP and central corneal thickness. Clinicians should consider factors other than ethnicity when choosing an ocular hypotensive medication for a patient. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00000125. PMID- 17420365 TI - Prophylaxis of acute posttraumatic bacterial endophthalmitis: a multicenter, randomized clinical trial of intraocular antibiotic injection, report 2. AB - OBJECTIVE: To evaluate the efficacy of intraocular gentamicin sulfate and clindamycin in the prevention of acute posttraumatic bacterial endophthalmitis following penetrating eye injuries. METHOD: We conducted a multicenter, randomized, double-masked controlled trial of 346 eyes with penetrating eye injury. Following primary repair, eyes were randomized to intracameral or intravitreal injection of 40 microg of gentamicin sulfate and 45 microg of clindamycin (cases) vs balanced salt solution (controls). MAIN OUTCOME MEASURE: Occurrence of endophthalmitis within 2 weeks. RESULTS: Endophthalmitis occurred in 8 (2.3%) of 167 eyes in the control group and only in 1 (0.3%) of 179 eyes in the case group (P = .04; odds ratio, 8.93 [95% confidence interval, 1.11-71.43]). In eyes with an intraocular foreign body, endophthalmitis developed in 7 of 25 control eyes and in none of 27 eyes receiving antibiotics. However, in eyes without an intraocular foreign body, endophthalmitis developed in 1 of 142 eyes and 1 of 152 eyes in the 2 groups, respectively (P value for interaction = .04). Intravitreal injection was superior to intracameral injection in preventing endophthalmitis (P value for interaction = .01). Vitreous culture results were positive in 6 (67%) of 9 eyes with endophthalmitis. CONCLUSION: Intraocular gentamicin and clindamycin are effective in the prevention of acute posttraumatic bacterial endophthalmitis in eyes with retained intraocular foreign body. APPLICATION TO CLINICAL PRACTICE: Prophylaxis of traumatic endophthalmitis. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00398658. PMID- 17420366 TI - Comparison of the modified Early Treatment Diabetic Retinopathy Study and mild macular grid laser photocoagulation strategies for diabetic macular edema. AB - OBJECTIVE: To compare 2 laser photocoagulation techniques for treatment of diabetic macular edema: the modified Early Treatment Diabetic Retinopathy Study (ETDRS) direct/grid photocoagulation technique and a potentially milder (but potentially more extensive) mild macular grid (MMG) laser technique in which microaneurysms are not treated directly and small mild burns are placed throughout the macula, whether or not edema is present. METHODS: Two hundred sixty-three subjects (mean age, 59 years) with previously untreated diabetic macular edema were randomly assigned to receive laser photocoagulation by either the modified ETDRS (162 eyes) or MMG (161 eyes) technique. Visual acuity, fundus photographs, and optical coherence tomography measurements were obtained at baseline and at 3.5, 8, and 12 months. Treatment was repeated if diabetic macular edema persisted. MAIN OUTCOME MEASURE: Change in optical coherence tomography measurements at 12-month follow-up. RESULTS: Among eyes with a baseline central subfield thickness of 250 microm or greater, central subfield thickening decreased by an average of 88 microm in the modified ETDRS group and by 49 microm in the MMG group at 12-month follow-up (adjusted mean difference, 33 microm; 95% confidence interval, 5-61 microm; P = .02). Weighted inner zone thickening by optical coherence tomography decreased by 42 microm in the modified ETDRS group and by 28 microm in the MMG group (adjusted mean difference, 14 microm; 95% confidence interval, 1-27 microm; P = .04); maximum retinal thickening (maximum thickening of the central and 4 inner subfields) decreased by 66 and 39 microm, respectively (adjusted mean difference, 27 microm; 95% confidence interval, 6-47 microm; P = .01), and retinal volume decreased by 0.8 and 0.4 mm3, respectively (adjusted mean difference, 0.3 mm3; 95% confidence interval, 0.02-0.53 mm3; P = .03). At 12 months, the mean change in visual acuity was 0 letters in the modified ETDRS group and 2 letters worse in the MMG group (adjusted mean difference, 2 letters; 95% confidence interval, -0.5 to 5 letters; P = .10). CONCLUSIONS: At 12 months after treatment, the MMG technique was less effective at reducing optical coherence tomography-measured retinal thickening than the more extensively evaluated current modified ETDRS laser photocoagulation approach. However, the visual acuity outcome with both approaches is not substantially different. Given these findings, a larger long-term trial of the MMG technique is not justified. APPLICATION TO CLINICAL PRACTICE: Modified ETDRS focal photocoagulation should continue to be a standard approach for treating diabetic macular edema. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00071773. PMID- 17420367 TI - Preventing posterior capsular opacification with an endocapsular equator ring in a young human eye: 2-year follow-up. AB - OBJECTIVE: To report the results of a specially designed closed ring with a square edge (endocapsular equator ring) in a young patient to prevent posterior capsular opacification. METHODS: One eye of a 22-year-old atopic patient underwent endocapsular equator ring implantation with a 1-piece polymethylmethacrylate intraocular lens immediately after phacoemulsification at Hara Eye Hospital on January 10, 2003. The solid flexible silicone ring has an outer diameter of 9.0 mm, is 1.0 mm wide and 1.0 mm thick, and has a square edge. The loops of the intraocular lens are fixed in the inner groove of the ring. The contralateral control eye underwent phacoemulsification and implantation with a conventional intraocular lens implantation. The Hayashi method was used to determine the posterior capsular opacification score. RESULTS: The ring retained the transparency of the entire posterior capsule. Two years postoperatively, the posterior capsular opacification score in the central area was 3.75 in the ring eye and 15.25 in the control eye, which underwent Nd:YAG laser capsulotomy 2.5 years postoperatively. CONCLUSION: An endocapsular equator ring effectively prevents posterior capsular opacification in a young patient with atopic cataracts. PMID- 17420368 TI - Predictive factors for corneal infiltrates with continuous wear of silicone hydrogel contact lenses. AB - OBJECTIVE: To estimate the cumulative probability and risk factors for developing corneal infiltrates after up to 3 years of continuous wear (CW) with lotrafilcon A lenses. METHODS: Patients were fitted with lotrafilcon A lenses and followed up for 3 years. The main outcome variable was the first occurrence of any infiltrative event in either eye. Cox proportional hazards regression was used to model the probability of developing infiltrates as a function of demographic and biomicroscopy findings. RESULTS: A total of 317 patients participated in this study. The Kaplan-Meier unadjusted cumulative incidence of a corneal infiltrate after CW was 5.7% (95% confidence interval [CI], 3.0%-8.4%) at the end of 1 year, 8.5% (95% CI, 5.2%-11.9%) at the end of 2 years, and 10.3% (95% CI, 6.6%-13.9%) at the end of 3 years. Corneal staining and limbal redness present in the affected eye on a previous visit were significantly associated with the development of an infiltrative event (hazard ratios, 7.23 and 3.18; P<.001 and P = .02, respectively). CONCLUSIONS: Corneal staining and limbal redness may predict the subsequent development of an infiltrative event among CW contact lens patients. The probability of remaining free of any corneal infiltrates at the end of 3 years of CW of contact lenses was 89.7% (95% CI, 86.1%-93.4%). PMID- 17420369 TI - Altered nitric oxide system in patients with open-angle glaucoma. AB - OBJECTIVE: To investigate the ocular blood flow response to systemic nitric oxide synthase inhibition in patients with primary open-angle glaucoma. METHODS: In 12 patients with glaucoma and 12 age-matched control subjects, subfoveal choroidal blood flow, optic nerve head blood flow, ocular fundus pulsation amplitude, intraocular pressure, and systemic hemodynamic parameters were measured at baseline and after inhibition of nitric oxide synthase by intravenous administration of NG-monomethyl-L-arginine. RESULTS: The increase in blood pressure in response to NG-monomethyl-L-arginine was comparable between the 2 study cohorts. In patients with glaucoma, the decrease of optic nerve head blood flow (P = .03) and fundus pulsation amplitude (P<.001) during nitric oxide synthase inhibition was significantly less pronounced than in healthy control subjects. A tendency toward a reduced response in choroidal blood flow was seen (P = .051 between groups) in patients with glaucoma. CONCLUSIONS: This is the first in vivo study providing evidence for an altered ocular L-arginine/nitric oxide system in patients with glaucoma. Normalization of the ocular nitric oxide production may be beneficial in terms of normalization of ocular blood flow and neuroprotection of retinal ganglion cells. PMID- 17420370 TI - Dengue-associated maculopathy. AB - OBJECTIVE: To describe the clinical spectrum of fundus manifestations and angiographic and optical coherence tomographic features of dengue-associated maculopathy in a large series. METHODS: We reviewed clinical records of patients diagnosed as having dengue maculopathy at the Singapore National Eye Centre between January 1, 2002, and December 31, 2005. RESULTS: We identified 41 patients with serological evidence of dengue fever who had ocular signs and symptoms not attributable to other diseases within 1 month after onset of symptoms of dengue. Seventy-one eyes had maculopathy. Mean best-corrected visual acuity in the affected eye was 20/40 (range, hand motions to 20/20). Intraretinal hemorrhages were seen in 45% of eyes, usually in association with venous sheathing. Fundus fluorescein angiography demonstrated venular occlusion in 25% or arteriolar and/or venular leakage in 3% and 13%, respectively. Yellow subretinal dots were an unusual finding in 28%. Of these, 50% showed corresponding hypofluorescent spots on indocyanine green angiography. Central or paracentral scotomas were observed in 63%. Twenty-eight patients received steroid treatment. Mean visual acuity showed significant improvement between weeks 2 and 4, with an increasing proportion of eyes achieving a best-corrected visual acuity of 20/40 or better across time. CONCLUSION: Fundus fluorescein and indocyanine green angiography, optical coherence tomography, and visual field testing are useful tools in the diagnosis of dengue maculopathy. PMID- 17420371 TI - MIRAgel: hydrolytic degradation and long-term observations. AB - OBJECTIVE: To analyze long-term complications of hydrogel (MIRAgel; MIRA Inc, Waltham, Mass) explants. DESIGN: Institutional clinical study of a retrospective, interventional case series of patients. We included 415 patients with complete reattachment of the retina 6 months after surgery and up-to-date follow-up. Patients underwent ophthalmological examination at each visit (mean follow-up, 187 months), and 6 underwent computed tomography and/or magnetic resonance imaging. Main outcome measures included the MIRAgel explant removal rate, clinical manifestations related to removal, interval from the start of discomfort to removal, mean time from implantation to removal, culture yield of the removed elements, results of histological examination of the capsule surrounding the removed explants (12 cases), and micro-Fourier transform infrared spectroscopic analysis results of 3 recovered explants. RESULTS: MIRAgel explant removal was necessary in 27 (6.5%) of 415 patients who received MIRAgel material and in 27 (7.6%) of 357 patients who had had it for 7 or more years. Clinical manifestations were related to swelling of the MIRAgel material, with a mean interval of 15 (range, 6-22) months from starting symptoms to removal. The infrared spectroscopic analysis demonstrated the presence of carboxylic groups in 3 recovered explants that had swollen considerably. CONCLUSION: Prompt removal of MIRAgel explants when discomfort starts should be considered to avoid increased incidence of complications. PMID- 17420372 TI - Estimation of systemic complement C3 activity in age-related macular degeneration. AB - OBJECTIVES: To determine the role of systemic complement activation in the pathogenesis of age-related macular degeneration and to examine whether serum C3a des Arg reflects systemic complement activation, independent of individual complement component levels. METHODS: Plasma complement C3a des Arg levels and a single nucleotide polymorphism at position 402 of the complement factor H gene (CFH) were determined in 3 groups of subjects: 42 subjects with early age-related maculopathy, 42 subjects with neovascular (wet) age-related macular degeneration, and a control group of 38 subjects with no clinical evidence of age-related changes at the macula. RESULTS: The median (range) of plasma complement C3a des Arg levels in the age-related maculopathy and neovascular age-related macular degeneration groups were 52.6 (2.8-198.1) ng/mL and 60.9 (3.1-173.1) ng/mL, respectively. The levels were significantly raised compared with the control group (n = 38), which had a median (range) plasma complement C3a des Arg level of 40.3 (6.1-81.7) ng/mL (analysis of variance, P = .02). The concentration of plasma C3a des Arg did not differ significantly between those with different CFH genotypes (P = .07). CONCLUSION: Systemic activation of the complement system may contribute to the pathogenesis of age-related macular degeneration independent of CFH polymorphism. CLINICAL RELEVANCE: The results of this study may be relevant to aiming new treatment strategies toward reducing systemic low-grade inflammation. PMID- 17420373 TI - The eyelid margin: a transitional zone for 2 epithelial phenotypes. AB - OBJECTIVE: To investigate the differentiation profile of the epithelium of the eyelid margin in the primate. METHODS: The expression of cytokeratins (CKs) CK1/10, CK4, CK14, CK5/8, and CK19; involucrin; connexin 43; filaggrin; and muscarinic receptor subtypes (m1-m5) on eyelid tissues from adult Macaca fascicularis (n = 3) was studied by immunohistochemistry. RESULTS: Cytokeratin 1/10, CK4, CK19, filaggrin, and connexin 43 expression varied across the epithelium of the mucocutaneous junction (MCJ). At the MCJ, CK4-positive cells overlapped basal CK14-expressing cells. The meibomian gland duct and the MCJ revealed layers of CK14-positive cells. Cytokeratin 5/8 was expressed in the basal layer of the epidermis and conjunctiva, while involucrin-positive cells were superficial. The m2 was expressed throughout the conjunctiva; m3 was found in the basal cells of the skin, conjunctiva, and alveolar epithelial cells of the meibomian glands; and m4 was expressed in the suprabasal layers of the skin. CONCLUSIONS: The eyelid margin epithelium is formed from 2 different epithelial cell subpopulations with specific but overlapping distributions. The meibomian gland duct or MCJ may be a site of conjunctival progenitor cells. CLINICAL RELEVANCE: This study provides a basis for understanding eyelid pathological conditions and eventually for developing methods for cellular reconstruction of the eyelid margin using expanded progenitor cells. PMID- 17420374 TI - Subclinical atherosclerotic cardiovascular disease and early age-related macular degeneration in a multiracial cohort: the Multiethnic Study of Atherosclerosis. AB - OBJECTIVE: To investigate the relationship of subclinical atherosclerotic cardiovascular disease (CVD) and its risk factors with age-related macular degeneration (AMD) in the Multiethnic Study of Atherosclerosis. METHODS: This study included 6176 white, black, Hispanic, and Chinese participants aged 44 to 84 years from 6 communities in the United States. Measurements of subclinical CVD were performed according to standardized protocols. Fundus images were graded using the Wisconsin Age-Related Maculopathy Grading System. RESULTS: In analyses controlled for age, sex, race/ethnicity, and study location, early AMD was associated with a higher serum high-density lipoprotein cholesterol level (odds ratio per 15 mg/dL, 1.16; 95% confidence interval, 1.01-1.36) and the presence of echolucent carotid artery plaque (odds ratio for present vs no plaque, 0.37; 95% confidence interval, 0.18-0.74) in the whole cohort. Interactions of race/ethnicity and early AMD were found for carotid intima-media thickness, increasing severity of maximum carotid artery stenosis, serum triglyceride level, subclinical CVD severity, and Agatston calcium score. CONCLUSION: Few associations were found between subclinical CVD and CVD risk factors with early AMD. The findings of associations of early AMD with some signs of subclinical atherosclerotic CVD are different among the 4 racial/ethnic groups, which suggests that care must be taken in generalizing from one racial/ethnic group to another. PMID- 17420375 TI - Economic impact of visual impairment and blindness in the United States. AB - OBJECTIVES: To assess the economic impact of excess medical and informal care and the health utility loss associated with visual impairment and blindness in adults aged 40 years and older in the United States. METHODS: Medical Expenditure Panel Survey data from 1996 to 2002 were pooled to estimate the relationship of visual impairment and blindness with total medical expenditures, components of expenditures, days of informal care received, and health utility. Estimates accounting for the complex sampling design were based on regressions including confounders such as comorbidities and demographics. The aggregate economic impact was estimated by projecting average individual effects to the population of individuals with blindness and visual impairment. RESULTS: Blindness and visual impairment were significantly associated with higher medical care expenditures, a greater number of informal care days, and a decrease in health utility. The home care component of expenditures was most affected by blindness. The aggregate annual economic impact included $5.5 billion spent for medical care and the value of informal care as well as a loss of more than 209,000 quality-adjusted life years. CONCLUSIONS: Visual impairment has a large effect on home care. Any economic analysis of prevention, treatment, and rehabilitation should account for the fraction of the annual monetary cost and loss of quality-adjusted life years that can be averted. PMID- 17420376 TI - One donor cornea for 3 recipients: a new concept for corneal transplantation surgery. AB - OBJECTIVE: To describe the use of a single donor corneal tissue in 3 patients with corneal pathologic conditions. METHODS: A donor corneal tissue was divided into 3 parts using a microkeratome and a trephine. The anterior lamellar disc was transplanted into a patient with macular corneal dystrophy using the automated lamellar therapeutic keratoplasty technique. The posterior lamellar disc was transplanted into a patient with pseudophakic bullous keratopathy using the Descemet stripping automated endothelial keratoplasty technique. The peripheral corneoscleral rim was used for limbal stem cell transplantation in a child with limbal stem cell deficiency. RESULTS: All surgical procedures were performed successfully. At 3 months, the best-corrected visual acuities achieved following automated lamellar therapeutic keratoplasty, Descemet stripping automated endothelial keratoplasty, and limbal stem cell transplantation were 20/60, 20/40, and 20/200, respectively. CONCLUSION: The advent of customized component corneal transplantation techniques may allow the use of 1 donor cornea to treat multiple patients. PMID- 17420377 TI - The ongoing battle against posterior capsular opacification. PMID- 17420381 TI - Aspergillus infection of the orbital apex masquerading as Tolosa-Hunt syndrome. PMID- 17420382 TI - Orbital rhabdomyosarcoma in Li-Fraumeni syndrome. PMID- 17420383 TI - Improvement of noninfectious uveitis with fumaric acid esters: results of a pilot study. PMID- 17420384 TI - Acetazolamide in the treatment of X-linked retinoschisis maculopathy. PMID- 17420385 TI - Gigantic waves in the tear film generated by bubbles from a large glaucoma bleb. PMID- 17420386 TI - Visualization of lipofuscin accumulation in Stargardt macular dystrophy by high resolution Fourier-domain optical coherence tomography. PMID- 17420387 TI - Hypopigmentary fundus changes seen with cutaneous vitiligo. PMID- 17420388 TI - Perioperative glycemic control and diabetic retinopathy. PMID- 17420390 TI - Optic disc puncture evaluation. PMID- 17420391 TI - Atypical retinoblastoma in Sotos syndrome (cerebral gigantism). PMID- 17420393 TI - Liquid crystal glasses: feasibility and safety of a new modality for treating amblyopia. PMID- 17420394 TI - The eyes, brain, and bones of Johann Sebastian Bach. PMID- 17420395 TI - The forecast for podcasts: sunny skies but not necessarily with clear visibility. PMID- 17420396 TI - Lenticulostriate-medullary artery anastomoses in moyamoya disease. PMID- 17420397 TI - To drive or not to drive: roles of the physician, patient, and state. PMID- 17420398 TI - When the patient knows best: a challenge in translating clinical trial evidence into practice. PMID- 17420399 TI - American Academy of Neurology position statement on physician reporting of medical conditions that may affect driving competence. PMID- 17420400 TI - The impact of neuropathic pain on health-related quality of life: review and implications. AB - A number of high-quality studies have recently been published that examine the association between neuropathic pain and health-related quality of life (HRQoL). The current review identified 52 such studies in patients with six neuropathic pain conditions associated with lesions of either the peripheral (postsurgical neuropathic pain associated with breast and amputation surgery, postherpetic neuralgia, and painful diabetic neuropathy) or central (poststroke pain, spinal cord injury pain, multiple sclerosis pain) nervous system. The results provide strong evidence that the presence and severity of neuropathic pain are associated with greater impairments in a number of important HRQoL domains. However, the evidence also indicates that this impact varies somewhat as a function of the HRQoL domain being considered and that different measures of HRQoL are differentially sensitive to the effects of neuropathic pain. The findings have important implications for the selection of HRQoL domains and measures to use in clinical trials and in clinical research on HRQoL in persons with neuropathic pain and suggest that a biopsychosocial (as opposed to a primarily biomedical) approach would be appropriate for understanding and treating neuropathic pain. PMID- 17420401 TI - Stereotypies in Rett syndrome: analysis of 83 patients with and without detected MECP2 mutations. AB - BACKGROUND: Hand stereotypies are considered a hallmark of Rett syndrome (RTT) and are usually described as symmetric movements at the midline. However, related pathologies may show the same type of involuntary movement. Furthermore, patients with RTT also have stereotypies with other localizations that are less well characterized. METHODS: We analyzed stereotypies in 83 patients with RTT, 53 with and 30 without a mutation detected in the MECP2 gene. Patients were observed and videotaped always by the same pediatric neurologist. Stereotypies were classified, and data were submitted to statistical analysis for comparison of mutation-positive and -negative patients and analysis of their evolution with the disease. RESULTS: All the patients showed hand stereotypies that coincided with or preceded the loss of purposeful hand movements in 62% of the patients with MECP2 mutations. The hair pulling stereotypy was more frequent in the group with detected mutations, whereas hand washing was not. Hand gaze was absent in all RTT patients with MECP2 mutations. Patients with MECP2 mutations also had more varied stereotypies, and the number of stereotypies displayed by each patient decreased significantly with age in this group. In all patients, stereotypies other than manual tended to disappear with the evolution of the disease. CONCLUSIONS: Although symmetric midline hand stereotypies were not specific to patients with an MECP2 mutation, some of the other stereotypies seemed to be more characteristic of this group. In patients younger than 10 years and meeting the necessary diagnostic criteria of Rett syndrome, the association of hand stereotypies without hand gaze, bruxism, and two or more of the other stereotypies seemed to be highly indicative of the presence of an MECP2 mutation. PMID- 17420402 TI - Quality of life outcomes of immediate or delayed treatment of early epilepsy and single seizures. AB - OBJECTIVE: To compare the impact of policies of immediate vs deferred treatment in patients with few or infrequent seizures on quality of life (QoL) outcomes. METHODS: We conducted a multicenter, randomized, unblinded study of immediate and deferred treatment. QoL data were collected by mail, using validated measures, for participants living in the UK and without major learning disability. Baseline questionnaires were returned by 441 adult patients; 333 returned 2-year follow-up questionnaires. This analysis is based on 331 patients (162 randomized to immediate, 169 to deferred treatment) returning both baseline and 2-year questionnaires. RESULTS: There were no significant differences at 2 years in QoL outcomes by treatment group. Patients randomized to deferred treatment were no more likely to report impairments in general health, cognitive function, psychological well-being, or social function. The one area of functioning affected was driving, where those randomized to deferred treatment were disadvantaged. There were clear QoL impacts both of taking antiepileptic drugs and, to an even greater extent, of continuing seizures. CONCLUSIONS: In treatment uncertain patients, there is a clear trade-off between adverse effects of seizures and adverse effects of taking antiepileptic drugs, i.e., neither policy examined in our study was associated with overall quality of life gains or losses longer term. PMID- 17420403 TI - Randomized, multicenter, dose-ranging trial of retigabine for partial-onset seizures. AB - OBJECTIVE: To evaluate the efficacy and safety of retigabine 600, 900, and 1,200 mg/day administered three times daily as adjunctive therapy in patients with partial-onset seizures. METHODS: A multicenter, randomized, double-blind, placebo controlled trial was performed. After an 8-week baseline phase, patients were randomized to a 16-week double-blind treatment period (8-week forced titration and 8-week maintenance) followed by either tapering or entry into an open-label extension study. Primary efficacy was the percentage change from baseline in monthly seizure frequency and compared across treatment arms. Secondary efficacy comparisons included the proportion of patients experiencing >/=50% reduction in seizure frequency (responder rate), emergence of new seizure types, and physician assessment of global clinical improvement. Safety/tolerability assessments included adverse events (AEs), physical and neurologic examinations, and clinical laboratory evaluations. Efficacy analyses were performed on the intent-to-treat population. RESULTS: Of the 399 randomized patients, 279 (69.9%) completed the double-blind treatment period. The median percent change in monthly total partial seizure frequency from baseline was -23% for 600 mg/day, -29% for 900 mg/day, and -35% for 1,200 mg/day vs -13% for placebo (p < 0.001 for overall difference across all treatment arms). Responder rates for retigabine were 23% for 600 mg/day, 32% for 900 mg/day (p = 0.021), and 33% for 1,200 mg/day (p = 0.016), vs 16% for placebo. The most common treatment-emergent AEs were somnolence, dizziness, confusion, speech disorder, vertigo, tremor, amnesia, abnormal thinking, abnormal gait, paresthesia, and diplopia. CONCLUSION: Adjunctive therapy with retigabine is well tolerated and reduces the frequency of partial onset seizures in a dose-dependent manner. PMID- 17420404 TI - 11C-PIB PET imaging in Alzheimer disease and frontotemporal lobar degeneration. AB - BACKGROUND: The PET tracer (11)C-labeled Pittsburgh Compound-B ((11)C-PIB) specifically binds fibrillar amyloid-beta (Abeta) plaques and can be detected in Alzheimer disease (AD). We hypothesized that PET imaging with (11)C-PIB would discriminate AD from frontotemporal lobar degeneration (FTLD), a non-Abeta dementia. METHODS: Patients meeting research criteria for AD (n = 7) or FTLD (n = 12) and cognitively normal controls (n = 8) underwent PET imaging with (11)C-PIB (patients and controls) and (18)F-fluorodeoxyglucose ((18)F-FDG) (patients only). (11)C-PIB whole brain and region of interest (ROI) distribution volume ratios (DVR) were calculated using Logan graphical analysis with cerebellum as a reference region. DVR images were visually rated by a blinded investigator as positive or negative for cortical (11)C-PIB, and summed (18)F-FDG images were rated as consistent with AD or FTLD. RESULTS: All patients with AD (7/7) had positive (11)C-PIB scans by visual inspection, while 8/12 patients with FTLD and 7/8 controls had negative scans. Of the four PIB-positive patients with FTLD, two had (18)F-FDG scans that suggested AD, and two had (18)F-FDG scans suggestive of FTLD. Mean DVRs were higher in AD than in FTLD in whole brain, lateral frontal, precuneus, and lateral temporal cortex (p < 0.05), while DVRs in FTLD did not significantly differ from controls. CONCLUSIONS: PET imaging with (11)C-labeled Pittsburgh Compound-B ((11)C-PIB) helps discriminate Alzheimer disease (AD) from frontotemporal lobar degeneration (FTLD). Pathologic correlation is needed to determine whether patients with PIB-positive FTLD represent false positives, comorbid FTLD/AD pathology, or AD pathology mimicking an FTLD clinical syndrome. PMID- 17420405 TI - Nocturnal blood pressure changes in patients with restless legs syndrome. AB - OBJECTIVES: To assess heart rate (HR) and blood pressure (BP) changes associated with periodic leg movements during sleep (PLMS) with or without EEG signs of arousal in subjects with primary restless legs syndrome (RLS). METHODS: Ten patients with RLS (4 women, aged 47.3 +/- 13.5 years) underwent one night of polysomnography along with noninvasive beat-to-beat BP monitoring. Ten PLMS with microarousals (PLMS-MA) and 10 PLMS without microarousals (PLMS-noMA) were analyzed in each subject. Systolic and diastolic BP (SBP, DBP) were measured within a 25-beat temporal window comprising 10 beats before and 15 beats after onset of each movement. PLMS-related BP changes were assessed by repeated measures one-way analysis of variance. BP changes associated with PLMS-MA and PLMS-noMA were compared by paired t-tests. Pearson correlation coefficients were used to assess the relationship between cardiovascular changes and clinical and polysomnographic variables. RESULTS: BP increased significantly in association with all PLMS (on average, SBP 22 mm Hg, DBP 11 mm Hg). BP changes associated with PLMS-MA were greater vs those associated with PLMS-noMA (p < 0.05). SBP and DBP changes increased with age and the duration of illness. CONCLUSIONS: Periodic leg movements-related repetitive nocturnal blood pressure fluctuations could contribute to the risk of cardiovascular diseases in patients with restless legs syndrome, especially in the elderly. PMID- 17420406 TI - Predicting a successful treatment in posterior canal benign paroxysmal positional vertigo. AB - OBJECTIVE: To elucidate the characteristics and prognostic value of positioning nystagmus during the second position of the Epley maneuver (90 degrees contralateral head turn from the initial Hallpike maneuver). METHOD: The Epley maneuver was performed in 126 patients with confirmed posterior canal benign paroxysmal positional vertigo (PC-BPPV). The characteristics of positioning nystagmus were investigated using video Frenzel goggles. RESULTS: During the second position, 99 patients developed torsional upbeating nystagmus, which was in the same direction (orthotropic nystagmus) as during the first position (Hallpike maneuver), whereas 15 patients showed a reversed pattern. In 12 patents, nystagmus was not induced during the second position. All 99 patients with orthotropic nystagmus had resolution of BPPV after the first or second trial of the Epley maneuver. In contrast, 12 of the 15 patients with reversed nystagmus and 8 of the 12 patients without nystagmus failed to resolve. CONCLUSION: During the second position of the Epley maneuver, an orthotropic pattern of nystagmus predicts a successful repositioning, whereas reversed nystagmus or no nystagmus is suggestive of poor response to repositioning. PMID- 17420407 TI - Physical activity and white matter lesion progression: assessment using MRI. AB - We evaluated the association between physical activity and changes in white matter lesions (WMLs) on MRI in a sample of 179 older adults comprising 59 incident cases of Alzheimer disease, 60 persons with mild cognitive impairment, and 60 persons who remained cognitively stable over a median 5-year follow-up. Physical activity was not significantly associated with a decreased rate of periventricular or deep WML progression. PMID- 17420408 TI - Head taps evoke a crossed vestibulo-ocular reflex. AB - Taps to the forehead on the midline, at the hairline (Fz), with a reflex hammer or powerful bone conduction vibrator caused short-latency surface potentials from beneath both eyes in all healthy subjects. The earliest negative responses were invariably absent from the eye contralateral to the side of a previous vestibular nerve section but were preserved despite sensorineural hearing loss. These responses probably reflect vestibular function via crossed otolith-ocular pathways. PMID- 17420409 TI - Intramedullary spinal cord abscess in a healthy woman. PMID- 17420410 TI - Solitary brainstem abscess. PMID- 17420411 TI - Tick-borne encephalitis with polyradiculitis documented by MRI. PMID- 17420412 TI - Cortical 5-HT1A receptor binding in patients with homozygous D90A SOD1 vs sporadic ALS. PMID- 17420413 TI - Reversible cognitive decline after DBS STN in PD and displacement of electrodes. PMID- 17420414 TI - Gray matter abnormalities in autism spectrum disorder revealed by T2 relaxation. PMID- 17420415 TI - Perforator stroke after elective stenting of symptomatic intracranial stenosis. PMID- 17420416 TI - Ten-year risk of dementia in subjects with mild cognitive impairment. PMID- 17420417 TI - How skeptical should we be about industry-sponsored studies? PMID- 17420418 TI - Erasmus Darwin (1731-1802): neurologist. PMID- 17420419 TI - Effect of cocoa and tea intake on blood pressure: a meta-analysis. AB - BACKGROUND: Epidemiological evidence suggests blood pressure-lowering effects of cocoa and tea. We undertook a meta-analysis of randomized controlled trials to determine changes in systolic and diastolic blood pressure due to the intake of cocoa products or black and green tea. METHODS: MEDLINE, EMBASE, SCOPUS, Science Citation Index, and the Cochrane Controlled Trials Register were searched from 1966 until October 2006 for studies in parallel group or crossover design involving 10 or more adults in whom blood pressure was assessed before and after receiving cocoa products or black or green tea for at least 7 days. RESULTS: Five randomized controlled studies of cocoa administration involving a total of 173 subjects with a median duration of 2 weeks were included. After the cocoa diets, the pooled mean systolic and diastolic blood pressure were -4.7 mm Hg (95% confidence interval [CI], -7.6 to -1.8 mm Hg; P = .002) and -2.8 mm Hg (95% CI, 4.8 to -0.8 mm Hg; P = .006) lower, respectively, compared with the cocoa-free controls. Five studies of tea consumption involving a total of 343 subjects with a median duration of 4 weeks were selected. The tea intake had no significant effects on blood pressure. The estimated pooled changes were 0.4 mm Hg (95% CI, 1.3 to 2.2 mm Hg; P = .63) in systolic and -0.6 mm Hg (95% CI, -1.5 to 0.4 mm Hg; P = .38) in diastolic blood pressure compared with controls. CONCLUSION: Current randomized dietary studies indicate that consumption of foods rich in cocoa may reduce blood pressure, while tea intake appears to have no effect. PMID- 17420420 TI - Insulin resistance and inflammation as precursors of frailty: the Cardiovascular Health Study. AB - BACKGROUND: Our research group has previously shown that the geriatric syndrome of frailty is associated with features of the metabolic syndrome (MetS) on cross sectional analysis. METHODS: To test whether MetS and its physiologic determinants-insulin resistance as measured by homeostasis model assessment score (IR-HOMA), increased inflammation and coagulation factor levels, and elevated blood pressure-are associated with incident frailty, we studied a subcohort of participants from the Cardiovascular Health Study observed from 1989/1990 through 1998/1999: 3141 community-dwelling adults, aged 69 to 74 years, without frailty and illnesses that increase inflammation markers or mimic frailty. The association of baseline MetS, IR-HOMA, levels of inflammation and coagulation factors, and systolic blood pressure (SBP) with time to onset of frailty was adjusted for demographic and psychosocial factors and incident events. Our main outcome measure was incident frailty. RESULTS: Metabolic syndrome was not significantly associated with incident frailty (hazard ratio, 1.16 (95% confidence interval [CI], 0.85-1.57). On the other hand, IR-HOMA and C-reactive protein levels were associated with incident frailty: for every standard deviation increment the hazard ratio for frailty was 1.15 (95% CI, 1.02-1.31) and 1.16 (95% CI, 1.02-1.32), respectively. The white blood cell count and factor VIIIc levels had a borderline association. Elevated systolic blood pressure had no association. Similar trends were found for incident prefrailty, a condition that precedes frailty. CONCLUSIONS: Two physiologic components of MetS- IR-HOMA and inflammation-are associated with incident frailty. Based on these results, IR HOMA can be considered part of a larger process that leads to generalized decline. PMID- 17420421 TI - Heterogeneity in the prevalence of risk factors for cardiovascular disease and type 2 diabetes mellitus in obese individuals: effect of differences in insulin sensitivity. AB - BACKGROUND: The possibility that substantial heterogeneity in metabolic abnormalities exists in moderately obese individuals has not been emphasized in studies of the effect of obesity on morbidity and mortality. We tested the hypothesis that risk factors for type 2 diabetes mellitus and cardiovascular disease vary dramatically in moderately obese individuals as a function of differences in a specific measure of insulin sensitivity. METHODS: Participants included 211 apparently healthy, obese (body mass index [calculated as weight in kilograms divided by height in meters squared], 30.0-34.9) volunteers for weight loss studies. Main outcome measures included insulin-mediated glucose uptake as quantified by the insulin suppression test and metabolic variables known to increase the risk for type 2 diabetes and cardiovascular disease. RESULTS: Insulin sensitivity varied 6-fold. When compared with the most insulin-sensitive third, the most insulin-resistant third of the population had significantly higher (P<.001) systolic and diastolic blood pressure (139 +/- 20 vs 123 +/- 18 mm Hg, and 83 +/- 3 vs 75 +/- 10 mm Hg, respectively), higher fasting and 2-hour oral glucose load concentrations (103 +/- 11 vs 95 +/- 11 mg/dL [5.7 +/- 0.6 vs 5.3 +/- 0.6 mmol/L], and 139 +/- 30 vs 104 +/- 19 mg/dL [7.7 +/- 1.7 vs 5.8 +/- 1.1 mmol/L], respectively), higher plasma triglyceride concentrations (198 +/- 105 vs 114 +/- 51 mg/dL [2.2 +/- 1.2 vs 1.3 +/- 0.6 mmol/L]), lower plasma high density lipoprotein cholesterol concentrations (41 +/- 9 vs 50 +/- 13 mg/dL [1.1 +/- 0.2 vs 1.3 +/- 0.3 mmol/L]), and more prevalent impaired glucose tolerance (47% vs 2%). CONCLUSIONS: The magnitude of risk factors for type 2 diabetes and cardiovascular disease varies markedly in moderately obese individuals as a function of differences in degree of insulin sensitivity. Because not all moderately obese individuals are at similar risk for developing type 2 diabetes and cardiovascular disease, intensive therapeutic interventions should be addressed to the insulin-resistant subset of this population. PMID- 17420422 TI - Physicians' observations and interpretations of the influence of religion and spirituality on health. AB - BACKGROUND: In spite of a substantial body of empirical data, professional disagreement persists regarding whether and how religion and spirituality (hereinafter "R/S" and treated as a single concept) influences health. This study examines the association between physicians' religious characteristics and their observations and interpretations of the influence of R/S on health. METHODS: A cross-sectional survey was mailed to a stratified, random sample of 2000 practicing US physicians from all specialties. Physicians were asked to estimate how often patients mention R/S issues, how much R/S influences health, and in what ways the influence is manifested. RESULTS: The response rate was 63%. Most physicians (56%) believed that R/S had much or very much influence on health, but few (6%) believed that R/S often changed "hard" medical outcomes. Rather, most physicians believed that R/S (1) often helps patients to cope (76%), (2) gives patients a positive state of mind (75%), and (3) provides emotional and practical support via the religious community (55%). Compared with those with low religiosity, physicians with high religiosity are substantially more likely to (1) report that patients often mention R/S issues (36% vs 11%)(P<.001); (2) believe that R/S strongly influences health (82% vs 16%) (P<.001); and (3) interpret the influence of R/S in positive rather than negative ways. CONCLUSION: Patients are likely to encounter quite different opinions about the relationship between their R/S and their health, depending on the religious characteristics of their physicians. PMID- 17420423 TI - Responding to a small-scale bioterrorist anthrax attack: cost-effectiveness analysis comparing preattack vaccination with postattack antibiotic treatment and vaccination. AB - BACKGROUND: In 2001, a small-scale bioterrorism-related anthrax attack was perpetrated via the US mail. The optimal future response may require strategies different from those required in a large-scale attack. METHODS: We conducted a cost-effectiveness analysis using Monte Carlo simulation during a 10-year time frame from a societal perspective to determine the optimal response strategy for a small-scale anthrax attack perpetrated against US Postal Service distribution centers in a large metropolitan area. Three strategies were compared: preattack vaccination of all US distribution center postal workers, postattack antibiotic therapy followed by vaccination of exposed personnel, and postattack antibiotic therapy without vaccination of exposed personnel. Outcome measures were costs, quality-adjusted life-years, and incremental cost-effectiveness. The probabilities for anthrax exposure and infection; vaccine and antibiotic benefits, risks, and costs; and associated clinical outcomes were derived from the medical literature and from bioterrorism experts. RESULTS: Postattack antibiotic therapy and vaccination of exposed postal workers is the most cost effective response compared with other strategies. The incremental cost effectiveness is $59 558 per quality-adjusted life-year compared with postattack antibiotic therapy alone. Preattack vaccination of all distribution center workers is less effective and more costly than the other 2 strategies. Assuming complete adherence to preattack vaccination, the incremental cost-effectiveness compared with postattack antibiotic therapy alone is almost $2.6 million per quality-adjusted life-year. CONCLUSION: Despite uncertainties about a future anthrax attack and exposure risk, postattack antibiotic therapy and vaccination of exposed personnel seems to be the optimal response to an attack perpetrated through the US Postal Service. PMID- 17420424 TI - Physician consideration of patients' out-of-pocket costs in making common clinical decisions. AB - BACKGROUND: Patients face growing cost-sharing through higher deductibles and other out-of-pocket (OP) expenses, with uncertain effects on clinical decision making. METHODS: We analyzed data on 6628 respondents to the nationally representative 2004-2005 Community Tracking Study Physician Survey to examine how frequently physicians report considering their insured patients' OP expenses when prescribing drugs, selecting diagnostic tests, and choosing inpatient vs outpatient care settings. Responses were dichotomized as always/usually vs sometimes/rarely/never. In separate multivariate logistic regressions, we examined associations between physicians' reported frequency of considering OP costs for each type of decision and characteristics of individual physicians and their practices. RESULTS: Seventy-eight percent of physicians reported routinely considering OP costs when prescribing drugs, while 51.2% reported doing so when selecting care settings, and 40.2% when selecting diagnostic tests. In adjusted analyses, primary care physicians were more likely than medical specialists to consider patients' OP costs in choosing prescription drugs (85.3% vs 74.5%) (P<.001), care settings (53.9% vs 43.1%) (P<.001), and diagnostic tests (46.3% vs 29.9%) (P<.001). Physicians working in large groups or health maintenance organizations were more likely to consider OP costs in prescribing generic drugs (P<.001 for comparisons with solo and 2-person practices), but those in solo or 2 person practices were more likely to do so in choosing tests and care settings (P<.05 for all comparisons with other practice types). Physicians providing at least 10 hours of charity care a month were more likely than those not providing any to consider OP costs in both diagnostic testing (40.7% vs 35.8%) (P<.001) and care setting decisions (51.4% vs 47.6%) (P<.005). CONCLUSION: Cost-sharing arrangements targeting patients are likely to have limited effects in safely reducing health care spending because physicians do not routinely consider patients' OP costs when making decisions regarding more expensive medical services. PMID- 17420425 TI - Permanent pacemaker and implantable cardioverter defibrillator infection: a population-based study. AB - BACKGROUND: The incidence of cardiac device infection is not well understood. Bloodstream infection (BSI) in patients with permanent pacemakers or implantable cardioverter-defibrillators (hereafter, defibrillators) may reflect device infection. METHODS: Retrospective, population-based cohort study of all adult patients with cardiac devices who resided in Olmsted County, Minnesota, from 1975 to 2004. The medical linkage-system of the Rochester Epidemiology Project and standardized criteria were used to identify all cases of BSI and device infection. The incidence of device infection was calculated with person-years of follow-up after device implantation. RESULTS: A total of 1524 patients with cardiac devices were included in the cohort. Total person-time of follow-up was 7578 years. The incidence of definite device infection was 1.9 per 1000 device years (95% confidence interval [CI], 1.1-3.1). The incidence of pocket infection without BSI was 1.37 per 1000 device-years (95% CI, 0.62-3.05), and pocket infection with BSI or device-related endocarditis 1.14 per 1000 device years (95% CI, 0.47-2.74). The cumulative probability of device infection was higher among patients with defibrillators compared with those with pacemakers, P<.001. Twelve (54.6%) of 22 cases of Staphylococcus aureus BSI had definite or possible cardiac device infection vs 3 (12.0%) of 25 cases of bloodstream infection due to gram negative bacilli (P = .004). CONCLUSIONS: To our knowledge, this is the first population-based study to describe the incidence of cardiac device infection. Device infection was common during episodes of S aureus BSI. The rate of cardiac device infection was higher in patients with defibrillators than in those with pacemakers. PMID- 17420426 TI - Variation in nursing home antipsychotic prescribing rates. AB - BACKGROUND: Excessive prescribing of antipsychotic therapy is a concern owing to their potential to cause serious adverse events. We explored variation in the use of antipsychotic therapy across nursing homes in Ontario, Canada, and determined if prescribing decisions were based on clinical indications. METHODS: A point prevalence study of antipsychotic therapy use in 47 322 residents of 485 provincially regulated nursing homes in December 2003. Facilities were classified into quintiles according to their mean antipsychotic prescribing rates. Residents were grouped into those with a potential clinical indication or no identified clinical indication for antipsychotic therapy. RESULTS: A total of 15 317 residents (32.4%) were dispensed an antipsychotic agent. The mean rate of antipsychotic prescribing by home ranged from 20.9% in the quintile of facilities with the lowest mean prescribing rates (quintile 1) to 44.3% in facilities with the highest mean prescribing rates (quintile 5). Compared with individuals residing in nursing homes with the lowest mean antipsychotic prescribing rates, those residing in facilities with the highest rates were 3 times more likely to be dispensed an antipsychotic agent (adjusted odds ratio [AOR], 3.0; 95% confidence interval [CI], 2.74-3.19). Similar rates were observed among residents with psychoses with or without dementia (AOR, 2.7; 95% CI, 2.35-3.09) and residents without psychoses or dementia (AOR, 2.9; 95% CI, 2.19-3.81) who had no identifiable indication for an antipsychotic therapy. CONCLUSION: Residents in facilities with high antipsychotic prescribing rates were about 3 times more likely than those in facilities with low prescribing rates to be dispensed an antipsychotic agent, irrespective of their clinical indication. PMID- 17420427 TI - Older age and the response to and tolerability of antiretroviral therapy. AB - BACKGROUND: The unique health needs of a growing older adult population infected with human immunodeficiency virus (HIV) require study, especially in terms of the response to and tolerability of highly active antiretroviral therapy (HAART). METHODS: Changes in HIV clinical markers after HAART initiation were compared among 2259 patients aged 18 to 39 years (reference group), 1834 patients aged 40 to 49 years, and 997 patients 50 years or older enrolled in an integrated health care system. RESULTS: Patients 50 years or older were more likely to achieve HIV RNA levels of less than 500 copies/mL within 1 year of HAART initiation (hazard ratio [HR], 1.15; P =.009), but adjustment for adherence attenuated this finding (HR, 1.03; P =.59). Subsequent HIV RNA level rebound (to > or =1000 copies/mL) was less likely among patients aged 40 to 49 years (HR, 0.81; P =.01), which persisted after adjustment for adherence (HR, 0.79; P =.004). In year 1 of HAART, younger patients had larger CD4 T-cell count increases (131.8, 121.3, and 111.8 CD4 T cells/microL per year among patients aged 18-39, 40-49, and > or =50 years, respectively; P =.046). In years 2 through 6, older patients had larger CD4 T cell count increases (4.5, 11.6, and 9.7 CD4 T cells/microL per year among patients aged 18-39, 40-49, and > or =50 years, respectively; P =.04). After adjustment for adherence, age differences in CD4 T-cell count changes remained in year 1 (P =.02) but not in years 2 through 6 (P =.08). Other factors, including comorbidities, had no effect on study results. Metabolic (glucose and lipids), hematologic (absolute neutrophils and hemoglobin), and renal (creatinine) abnormalities were more likely among older patients. CONCLUSION: Despite a higher risk of adverse events, patients 50 years or older sustained high therapy adherence to maintain improved virological outcomes and to compensate for their early blunted CD4 T-cell count response compared with younger patients. PMID- 17420428 TI - Success of clinical care for human immunodeficiency virus infection according to demographic group among sexually infected patients in a routine clinic population, 1999 to 2004. AB - BACKGROUND: The success of clinical care for human immunodeficiency virus infection may vary across demographic groups, because of patient- and health care related factors. METHODS: A total of 2386 patients sexually infected with the human immunodeficiency virus were seen in a London clinic from July 1, 1999, to December 31, 2004. We examined demographic variation and trends over time in the prevalence of the following: (1) a CD4 cell count of 200/microL or less; (2) a viral load of greater than 50 copies/mL among patients receiving antiretroviral therapy (ART); and (3) a viral load of greater than 50 copies/mL among patients receiving ART for 24 weeks or longer. RESULTS: Subjects were homosexual men (63.1%), white heterosexual men (4.3%) and women (5.1%), and black African or other ethnicity heterosexual men (10.2%) and women (17.3%). The CD4 cell count at the first clinic visit was highest among homosexual men and lowest among black African heterosexual men. From 1999 to 2004, ART use increased from 61.9% to 75.5%. The prevalence of a CD4 cell count of 200/microL or less decreased from 19.6% to 9.0%. The prevalence of a viral load of greater than 50 copies/mL decreased from 36.9% to 14.5% among patients receiving ART, and from 31.2% to 10.1% among patients receiving ART for 24 weeks or longer. Demographic variation in the prevalence of each outcome was apparent among men throughout the period: homosexual men had the most favorable profile, and black African heterosexual men had the least favorable profile. Differences were much greater for low CD4 cell count than for raised viral load while receiving ART. There was no consistent demographic variation among women. Favorable trends over time occurred within each demographic group, and were as strong among black African patients as among other subgroups. CONCLUSIONS: The success of clinical care for human immunodeficiency virus infection increased substantially from 1999 to 2004 in this routine clinic population. All demographic subgroups benefited from improvements, despite ongoing differences in the prevalence of immunosuppression. PMID- 17420429 TI - Albuminuria and renal function in homozygous sickle cell disease: observations from a cohort study. AB - BACKGROUND: The glomerular filtration rate (GFR) in homozygous sickle cell (SS) disease is supranormal in childhood but falls steeply with age, often culminating in renal failure. The risk factors underlying these observations are unclear. We therefore sought to investigate the relationships between blood pressure, renal hemodynamics, and urinary albumin excretion in subjects with SS disease and matched controls with a normal AA genotype (hereinafter, controls) as a prelude to intervention studies. METHODS: Serum creatinine level, GFR, effective renal plasma flow, blood pressure, and urinary albumin and creatinine excretion rates were measured in Jamaican individuals with SS disease aged 18 to 23 years and in controls followed from birth in a cohort study. RESULTS: Compared with controls, subjects with SS disease showed lower blood pressure and normal or supranormal GFR and effective renal plasma flow. Urinary albumin excretion exceeded 20 microg/min in 26% of subjects with SS disease and correlated positively with GFR and systolic blood pressure and negatively with hematocrit. A higher GFR and increased tubular secretion of creatinine combined to lower serum creatinine levels in patients with SS disease, giving an upper limit of the reference range of 0.90 mg/dL (80 micromol/L) in men and 0.77 mg/dL (68 micromol/L) in women. In addition, creatinine clearance measurements were consistently greater than GFR in subjects with SS disease. CONCLUSIONS: The GFR remained within reference range or elevated in patients with SS disease aged 18 to 23 years. The higher GFR in patients with albuminuria was consistent with the hypothesis that high glomerular flows cause renal damage. Lower serum creatinine levels characterize patients with SS disease, and a revised clinical definition based on serum creatinine level alone is proposed. PMID- 17420430 TI - A predictive model of recurrent lower extremity cellulitis in a population-based cohort. AB - BACKGROUND: Cellulitis is common and recurs in some patients. The study described herein derived a predictive model for the recurrence of lower extremity cellulitis in a population-based cohort. METHODS: We conducted a retrospective, population-based cohort study using the Rochester Epidemiology Project. We reviewed the medical records of Olmsted County, Minnesota, residents with lower extremity cellulitis occurring from January 1, 1999, to June 30, 2000. Univariate and multivariate Cox proportional hazards analyses were performed to evaluate risk factors in patients who experienced recurrent lower extremity cellulitis within 2 years. A predictive model was developed to estimate risk of recurrence based on a score of risk factors identified by multivariate analysis. RESULTS: A total of 209 episodes met the definition of lower extremity cellulitis. Thirty five patients (16.7%) experienced recurrence within 2 years. Multivariate analysis identified tibial area involvement, prior malignancy, and dermatitis affecting the ipsilateral limb as independent risk factors for recurrence, with hazard ratios of 5.02, 3.87, and 2.99 (P<.01), respectively. A score calculated from these variables (a count of 0, 1, 2, or 3) was developed to measure risk of recurrence. Based on the predictive model, the estimated probability of recurrence (95% confidence interval [CI]) within 2 years was 5.0% (95% CI, 1.6% 8.2%), 17.3% (95% CI, 11.1%-23.0%), 50.6% (95% CI, 34.2%-63.0%), or 92.8% (95% CI, 51.9%-98.9%) for a score of 0, 1, 2 or 3, respectively. CONCLUSIONS: We derived a model including tibial area involvement, history of cancer, and dermatitis to predict recurrence of lower extremity cellulitis. Potential interventions can be incorporated into treatment to diminish the proclivity for recurrence in high-risk patients. PMID- 17420431 TI - Smoking status as a clinical indicator for alcohol misuse in US adults. AB - BACKGROUND: Screening for alcohol use in primary care settings is recommended by clinical care guidelines but is not adhered to as strongly as screening for smoking. It has been proposed that smoking status could be used to enhance the identification of alcohol misuse in primary care and other medical settings, but national data are lacking. Our objective was to investigate smoking status as a clinical indicator for alcohol misuse in a national sample of US adults, following clinical care guidelines for the assessment of these behaviors. METHODS: Analyses are based on a sample of 42 374 US adults from the National Epidemiological Survey on Alcohol and Related Conditions (Wave I, 2001-2002). Odds ratios (ORs), 95% confidence intervals (CIs), and test characteristics (sensitivity, specificity, positive and negative predictive values, and positive likelihood ratio of smoking behavior [daily, occasional, or former]) were determined for the detection of hazardous drinking behavior and alcohol-related diagnoses, assessed by the Alcohol Use Disorder and Associated Disabilities Interview Schedule-IV. RESULTS: Daily, occasional, and ex-smokers were more likely than never smokers to be hazardous drinkers (OR, 3.23 [95% CI, 3.02-3.46]; OR, 5.33 [95% CI, 4.70-6.04]; OR, 1.19 [95% CI, 1.10-1.28], respectively). Daily and occasional smokers were more likely to meet criteria for alcohol diagnoses (OR, 3.52 [95% CI, 3.19-3.90] and OR, 5.39 [95% CI, 4.60-6.31], respectively). For the detection of hazardous drinking by current smoking (occasional smokers + daily smokers), sensitivity was 42.5%; specificity, 81.9%; positive predictive value, 45.3% (vs population rate of 26.1%); and positive likelihood ratio, 2.34. For the detection of alcohol diagnoses by current smoking, sensitivity was 51.4%; specificity, 78.0%; positive predictive value, 17.8% (vs population rate of 8.5%); and positive likelihood ratio, 2.33. CONCLUSIONS: Occasional and daily smokers were at heightened risk for hazardous drinking and alcohol use diagnoses. Smoking status can be used as a clinical indicator for alcohol misuse and as a reminder for alcohol screening in general. PMID- 17420432 TI - Serum ferritin and transferrin saturation in Asians and Pacific Islanders. AB - BACKGROUND: Asians and Pacific Islanders in the Hemochromatosis and Iron Overload Screening (HEIRS) Study had the highest prevalence of elevated serum ferritin (SF) and transferrin saturation (TS) levels, but to our knowledge, the reasons for this have not been investigated. METHODS: Using multiple linear regression, we compared TS and SF distributions for 42 720 Asian, Pacific Islander, and white HEIRS Study participants recruited through 5 field centers in North America who did not have HFE C282Y or H63D alleles. RESULTS: Compared with their white counterparts, Asian men had a 69-ng/mL (155-pmol/L) higher adjusted mean SF level and a 3% higher TS level (P<.001); Asian women had 23-ng/mL (52-pmol/L) higher adjusted mean SF level and a 3% higher TS level (P<.001). The mean TS level of Asian women was higher than that of Pacific Islander women, and the mean SF level of Pacific Islander men was significantly higher than that of white men. These differences remained significant after adjusting for self-reported history of diabetes or liver disease. Additional information for selected participants suggested that these differences are largely unrelated to mean corpuscular volume less than 80 fL, body mass index, or self-reported alcohol intake. Available liver biopsy and phlebotomy data indicated that iron overload is probably uncommon in Asian participants. CONCLUSION: Higher TS and SF levels in persons of Asian or Pacific Island heritage may need to be interpreted differently than for whites, although the biological basis and clinical significance of higher levels among Asians and Pacific Islanders are unclear. PMID- 17420433 TI - The spectrum of community-based hospitalist practice: a call to tailor internal medicine residency training. PMID- 17420434 TI - The long and short of metformin-related vitamin B12 deficiency. PMID- 17420435 TI - Vitamin B12 deficiency in patients receiving metformin: clinical data. PMID- 17420436 TI - Metformin and vitamin B12 deficiency: association and not causation may play a major role in cohort studies. PMID- 17420437 TI - Acute vs chronic secondhand smoke exposure. PMID- 17420438 TI - Avoiding the regulatory capture of the Food and Drug Administration. PMID- 17420439 TI - Sweeping changes to Food and Drug Administration and drug safety regulations. PMID- 17420440 TI - Perfectionism and the cortisol response to psychosocial stress in men. AB - OBJECTIVE: To test the hypothesis that perfectionism is an important moderator of the neuroendocrine stress response, with higher perfectionism predicting increased neuroendocrine activation. METHODS: A total of 50 middle-aged men underwent an acute standardized psychosocial stress task (Trier Social Stress Test). Perfectionism, cognitive appraisal of the stressful situation, trait anxiety, and various personality characteristics were assessed with questionnaires. Salivary cortisol, plasma epinephrine and norepinephrine, blood pressure, and heart rate were analyzed before and after stress. Circadian profiles of cortisol secretion during the day and in response to awakening were analyzed to assess basal activity of the hypothalamus-pituitary-adrenal (HPA) axis. Multiple regression analyses were conducted to identify predictors of the neuroendocrine stress response. RESULTS: Perfectionism was significantly associated with area under the total response curve with respect to increase (AUCi) of cortisol (r = 0.322, p = .046), but not with AUCi of norepinephrine (r = -0.217, p = .152) or AUCi of epinephrine (r = 0.116, p = .477). Hence, AUCi of cortisol was the main criterion. As possible predictors, trait anxiety, neuroticism, vital exhaustion, secondary appraisal, depression, and openness were considered. Regression analyses demonstrated that only perfectionism (beta = 0.45, p = .002) and secondary appraisal (beta = 0.50, p = .001) were independent predictors of AUCi of cortisol, the final model explaining 45% of the total variance in cortisol response (R2 = 0.45, "shrunken" R2 [sR2] = 0.38); perfectionism alone accounted for 18% of this variance (deltaR2 = 0.18, sR2 = 0.19). CONCLUSION: The typical cognitions, and presumably the associated emotions, of perfectionists seem to contribute independently to stress-induced bodily responses, including HPA axis activation, in response to psychosocial stress. PMID- 17420441 TI - Changes in depressive symptoms and glycemic control in diabetes mellitus. AB - OBJECTIVE: To investigate if changes in depressive symptoms would be associated with changes in glycemic control over a 12-month period in patients with Type 1 and Type 2 diabetes. METHODS: Ninety (Type 1 diabetes, n = 28; Type 2 diabetes, n = 62) patients having Beck Depression Inventory (BDI) levels of >10 were enrolled in the study. Of those 90 patients, 65 patients completed a 12-week cognitive behavioral therapy intervention. BDI was assessed at baseline and thereafter biweekly during 12 months. Hemoglobin (HbA1c) and fasting blood glucose levels were assessed at baseline and at four quarterly in-hospital follow-up visits. Linear mixed-model analysis was applied to determine the effects of time and diabetes type on depressive symptoms, HbA1c levels, and fasting glucose levels. RESULTS: Mean and standard deviation baseline BDI and HbA1c levels were 17.9 +/- 5.8 and 7.6 +/- 1.6, respectively, with no significant difference between patients with Type 1 and Type 2 diabetes. Mixed-model regression analysis found no difference between the groups with Type 1 and Type 2 diabetes in the within subject effect of BDI score on HbA1c or fasting glucose levels during the study. Depressive symptoms decreased significantly (p = .0001) and similarly over a 12 month period in both patients with Type 1 and Type 2 diabetes, whereas HbA1c and fasting glucose levels did not change significantly over time in either group. CONCLUSION: Changes in depressive symptoms were not associated with changes in HbA1c or fasting glucose levels over a 1-year period in either patients with Type 1 or Type 2 diabetes. PMID- 17420442 TI - Tonic and phasic heart rate as predictors of posttraumatic stress disorder. AB - OBJECTIVE: To examine the relationship between acute measures of a) heart rate (HR) immediately after traumatic injury, b) tonic (resting) HR at 1 week post injury, c) phasic (aroused) HR at 1 week post injury, and d) somatic symptoms of arousal in the prediction of subsequent posttraumatic stress disorder (PTSD). Fear conditioning models propose that HR reactivity shortly after trauma may predict PTSD. METHOD: In a longitudinal study, consecutive injury survivors (n = 197) admitted to a hospital trauma service were assessed within 1 week and at 12 months post injury. HR was assessed by paramedics at the site of the trauma and pulse oximetry technology at 1 week post trauma. Somatic symptoms of arousal were measured using the somatic scale on the Beck Anxiety Inventory (BAI). PTSD was assessed using the Clinician Administered PTSD Scale at 12 months. RESULTS: At 12 months post injury, PTSD was diagnosed in 10% of participants. Only HR change scores (phasic - tonic HR) and BAI scores significantly predicted later PTSD. CONCLUSIONS: These findings question the clinical usefulness of tonic HR as a biological marker of later PTSD. The finding that HR reactivity (phasic - tonic) predicts later PTSD has theoretical importance. The strongest predictor of later PTSD was somatic arousal. PMID- 17420443 TI - Benefit-finding in the cancer caregiving experience. AB - OBJECTIVE: To characterize the domains of benefit finding in caregiving among close family members of cancer survivors and to relate the domains of benefit finding in cancer caregiving to other psychosocial variables. Although cancer is a problem that involves the entire family, little is known about which caregivers of cancer survivors are likely to find meaning in their caregiving experience or about the relationship of benefit finding to caregivers' psychological adjustment. METHODS: A total of 896 family caregivers participated in the American Cancer Society Quality of Life Survey for Caregivers. Participating caregivers were primarily middle-aged (mean age = 54 years), educated (76% received > high school degree), and affluent (69% earned > or = $40,000 annual household income). RESULTS: Six domains of benefit finding in caregiving were identified by principal axis factor analysis: acceptance, empathy, appreciation, family, positive self-view, and reprioritization. The comparison between six- and one-domain models of benefit finding demonstrated the superiority of the six domain model. A series of hierarchical regression analyses showed that the domains of benefit finding were uniquely associated with life satisfaction and depression and that not all aspects of benefit finding in caregiving related to better adjustment. Specifically, coming to accept what happened and appreciating new relationships with others related to greater adaptation. Becoming more empathetic toward others and reprioritizing values related to greater symptoms of depression. CONCLUSIONS: These findings suggest that caregivers may benefit from interventions that enhance their ability to accept their situation and find meaning in their caregiving experience, which may improve their satisfaction with life and reduce their depressive symptoms. PMID- 17420444 TI - Use of social words in autobiographies and longevity. AB - OBJECTIVE: To analyze the relationship between social word use in autobiographies and longevity. Although there is substantial evidence that our social relationships are associated with mortality, interpretation of this work is weakened by the limitations of assessing the social environment with structured questionnaires and interviews. By analyzing the word content of autobiographies, we could assess spontaneous indicators of important social relationships and relate them to longevity. This technique is less subject to social desirability reporting biases and more sensitive to aspects of the social environment that are central to how one experiences his or her social world. METHODS: The autobiographies of 96 psychologists and 220 literary writers were digitized and scanned for social relationship word frequency via a computerized word counting program. Archival data were collected on birth and death dates, year of publication, place of birth, age when the autobiography was written, and sex. RESULTS: After controlling for sex, year of birth, and age at the time of writing, we found that higher use of words indicating social roles/integration (e.g., father, sister, neighbor, co-worker) was associated with an increased lifespan in both samples. Specific social categories assessing the use of family role terms (e.g., aunt, family, brother) and references to other individuals (e.g., they, we, us, everyone) also predicted longer life, but only in the sample of psychologists. CONCLUSIONS: Assessing social word use in autobiographies provided an indirect measure of social relationships that predicted longevity. This technique of analyzing writing samples may be useful in future archival research as well as in studies where it is desirable to study social relationships in an indirect fashion. PMID- 17420445 TI - A conserved phosphatase cascade that regulates nuclear membrane biogenesis. AB - A newly emerging family of phosphatases that are members of the haloacid dehalogenase superfamily contains the catalytic motif DXDX(T/V). A member of this DXDX(T/V) phosphatase family known as Dullard was recently shown to be a potential regulator of neural tube development in Xenopus [Satow R, Chan TC, Asashima M (2002) Biochem Biophys Res Commun 295:85-91]. Herein, we demonstrate that human Dullard and the yeast protein Nem1p perform similar functions in mammalian cells and yeast cells, respectively. In addition to similarity in primary sequence, Dullard and Nem1p possess similar domains and show similar substrate preferences, and both localize to the nuclear envelope. Additionally, we show that human Dullard can rescue the aberrant nuclear envelope morphology of nem1Delta yeast cells, functionally replacing Nem1p. Finally, Nem1p, has been shown to deposphorylate the yeast phosphatidic acid phosphatase Smp2p [Santos Rosa H, Leung J, Grimsey N, Peak-Chew S, Siniossoglou S (2005) EMBO J 24:1931 1941], and we show that Dullard dephosphorylates the mammalian phospatidic acid phosphatase, lipin. Therefore, we propose that Dullard participates in a unique phosphatase cascade regulating nuclear membrane biogenesis, and that this cascade is conserved from yeast to mammals. PMID- 17420446 TI - Regulation of CNS synapses by neuronal MHC class I. AB - Until recently, neurons in the healthy brain were considered immune-privileged because they did not appear to express MHC class I (MHCI). However, MHCI mRNA was found to be regulated by neural activity in the developing visual system and has been detected in other regions of the uninjured brain. Here we show that MHCI regulates aspects of synaptic function in response to activity. MHCI protein is colocalized postsynaptically with PSD-95 in dendrites of hippocampal neurons. In vitro, whole-cell recordings of hippocampal neurons from beta2m/TAP1 knockout (KO) mice, which have reduced MHCI surface levels, indicate a 40% increase in mini-EPSC (mEPSC) frequency. mEPSC frequency is also increased 100% in layer 4 cortical neurons. Similarly, in KO hippocampal cultures, there is a modest increase in the size of presynaptic boutons relative to WT, whereas postsynaptic parameters (PSD-95 puncta size and mEPSC amplitude) are normal. In EM of intact hippocampus, KO synapses show a corresponding increase in vesicles number. Finally, KO neurons in vitro fail to respond normally to TTX treatment by scaling up synaptic parameters. Together, these results suggest that postsynaptically localized MHCl acts in homeostatic regulation of synaptic function and morphology during development and in response to activity blockade. The results also imply that MHCI acts retrogradely across the synapse to translate activity into lasting change in structure. PMID- 17420447 TI - Phosphorylation-dependent regulation of nuclear localization and functions of integrin-linked kinase. AB - Integrin-linked kinase (ILK) is a phosphorylated protein that regulates physiological processes that overlap with those regulated by p21-activated kinase 1 (PAK1). Here we report the possible role of ILK phosphorylation by PAK1 in ILK mediated signaling and intracellular translocation. We found that PAK1 phosphorylates ILK at threonine-173 and serine-246 in vitro and in vivo. Depletion of PAK1 decreased the levels of endogenous ILK phosphorylation in vivo. Mutation of PAK1 phosphorylation sites on ILK to alanine reduced cell motility and cell proliferation. Biochemical fractionation, confocal microscopy, and chromatin-interaction analyses of human cells revealed that ILK localizes predominantly in the cytoplasm but also resides in the nucleus. Transfection of MCF-7 cells with point mutants ILK-T173A, ILK-S246A, or ILK-T173A; S246A (ILK-DM) altered ILK localization. Selective depletion of PAK1 dramatically increased the nuclear and focal point accumulation of ILK, further demonstrating a role for PAK1 in ILK translocation. We also identified functional nuclear localization sequence and nuclear export sequence motifs in ILK, delineated an apparently integral role for ILK in maintaining normal nuclear integrity, and established that ILK interacts with the regulatory region of the CNKSR3 gene chromatin to negatively modulate its expression. Together, these results suggest that ILK is a PAK1 substrate, undergoes phosphorylation-dependent shuttling between the cell nucleus and cytoplasm, and interacts with gene-regulatory chromatin. PMID- 17420448 TI - Visiting order on membrane proteins by using nanotechnology. PMID- 17420449 TI - Identity of a pathway for saccadic suppression. AB - Neurons in the superficial gray layer (SGS) of the superior colliculus receive visual input and excite intermediate layer (SGI) neurons that play a critical role in initiating rapid orienting movements of the eyes, called saccades. In the present study, two types of experiments demonstrate that a population of SGI neurons gives rise to a reciprocal pathway that inhibits neurons in SGS. First, in GAD67-GFP knockin mice, GABAergic SGI neurons that expressed GFP fluorescence were injected with the tracer biocytin to reveal their axonal projections. Axons arising from GFP-positive neurons in SGI terminated densely in SGS. Next, SGI neurons in rats and mice were stimulated by using the photolysis of caged glutamate, and in vitro whole-cell patch-clamp recordings were used to measure the responses evoked in SGS cells. Large, synaptically mediated outward currents were evoked in SGS neurons. These currents were blocked by gabazine, confirming that they were GABA(A) receptor-mediated inhibitory postsynaptic currents. This inhibitory pathway from SGI transiently suppresses visual activity in SGS, which in turn could have multiple effects. These effects could include reduction of perceptual blurring during saccades as well as prevention of eye movements that might be spuriously triggered by the sweep of the visual field across the retina. PMID- 17420450 TI - Direct and adaptor-mediated substrate recognition by an essential AAA+ protease. AB - Regulated proteolysis is required to execute many cellular programs. In Caulobacter crescentus, timely degradation of the master regulator CtrA by ClpXP protease is essential for cell-cycle progression and requires the colocalization of CtrA and RcdA. Here, we establish a biochemical framework to understand regulated proteolysis in C. crescentus and show that RcdA is not an adaptor for CtrA degradation. CtrA is rapidly degraded without RcdA and is recognized with an affinity comparable with the best ClpXP substrates. In contrast, SspBalpha, the alpha-proteobacterial homolog of SspB, functions as an adaptor to enhance degradation of specific substrates. Cargo-free SspBalpha is also itself a substrate of ClpXP-mediated proteolysis. Thus, our analysis (i) reveals the consequences of both direct and adaptor-stimulated recognition in mediating substrate specificity in vitro, (ii) reveals a potential regulatory role of controlled adaptor stability, and (iii) suggests that cell-cycle regulation of CtrA stability depends on repression of its intrinsic degradation rather than adaptor-mediated enhancement. PMID- 17420451 TI - Analysis of PALB2/FANCN-associated breast cancer families. AB - No more than approximately 30% of hereditary breast cancer has been accounted for by mutations in known genes. Most of these genes, such as BRCA1, BRCA2, TP53, CHEK2, ATM, and FANCJ/BRIP1, function in DNA repair, raising the possibility that germ line mutations in other genes that contribute to this process also predispose to breast cancer. Given its close relationship with BRCA2, PALB2 was sequenced in affected probands from 68 BRCA1/BRCA2-negative breast cancer families of Ashkenazi Jewish, French Canadian, or mixed ethnic descent. The average BRCAPRO score was 0.58. A truncating mutation (229delT) was identified in one family with a strong history of breast cancer (seven breast cancers in three female mutation carriers). This mutation and its associated breast cancers were characterized with another recently reported but unstudied mutation (2521delA) that is also associated with a strong family history of breast cancer. There was no loss of heterozygosity in tumors with either mutation. Moreover, comparative genomic hybridization analysis showed major similarities to that of BRCA2 tumors but with some notable differences, especially loss of 18q, a change that was previously unknown in BRCA2 tumors and less common in sporadic breast cancer. This study supports recent observations that PALB2 mutations are present, albeit not frequently, in breast cancer families. The apparently high penetrance noted in this study suggests that at least some PALB2 mutations are associated with a substantially increased risk for the disease. PMID- 17420452 TI - Ancient DNA reveals lack of postglacial habitat tracking in the arctic fox. AB - How species respond to an increased availability of habitat, for example at the end of the last glaciation, has been well established. In contrast, little is known about the opposite process, when the amount of habitat decreases. The hypothesis of habitat tracking predicts that species should be able to track both increases and decreases in habitat availability. The alternative hypothesis is that populations outside refugia become extinct during periods of unsuitable climate. To test these hypotheses, we used ancient DNA techniques to examine genetic variation in the arctic fox (Alopex lagopus) through an expansion/contraction cycle. The results show that the arctic fox in midlatitude Europe became extinct at the end of the Pleistocene and did not track the habitat when it shifted to the north. Instead, a high genetic similarity between the extant populations in Scandinavia and Siberia suggests an eastern origin for the Scandinavian population at the end of the last glaciation. These results provide new insights into how species respond to climate change, since they suggest that populations are unable to track decreases in habitat avaliability. This implies that arctic species may be particularly vulnerable to increases in global temperatures. PMID- 17420453 TI - In vivo selection for metastasis promoting genes in the mouse. AB - Here, we report the identification of a metastasis promoting factor by a forward genetic screen in mice. A retroviral cDNA library was introduced into the nonmetastatic cancer cell line 168FARN, which was then orthotopically transplanted into mouse mammary fat pads, followed by selection for cells that metastasize to the lung. The genes encoding the disulfide isomerase ERp5 and beta catenin were found to promote breast cancer invasion and metastasis. Disulfide isomerases (thiol isomerases), which catalyze disulfide bond formation, reduction, and isomerization, have not previously been implicated in cancer cell signaling and tumor metastasis. Overexpression of ERp5 promotes both in vitro migration and invasion and in vivo metastasis of breast cancer cells. These effects were shown to involve activation of ErbB2 and phosphoinositide 3-kinase (PI3K) pathways through dimerization of ErbB2. Activation of ErbB2 and PI3K subsequently stimulates RhoA and beta-catenin, which mediate the migration and invasion of tumor cells. Inhibition of ErbB2 and PI3K reverses the phenotypes induced by ERp5. Finally, ERp5 was shown to be up-regulated in human surgical samples of invasive breast cancers. These data identify a link between disulfide isomerases and tumor development, and provide a mechanism that modulates ErbB2 and PI3K signaling in the promotion of cancer progression. PMID- 17420454 TI - Genesis of hexavalent chromium from natural sources in soil and groundwater. AB - Naturally occurring Cr(VI) has recently been reported in ground and surface waters. Rock strata rich in Cr(III)-bearing minerals, in particular chromite, are universally found in these areas that occur near convergent plate margins. Here we report experiments demonstrating accelerated dissolution of chromite and subsequent oxidation of Cr(III) to aqueous Cr(VI) in the presence of birnessite, a common manganese mineral, explaining the generation of Cr(VI) by a Cr(III) bearing mineral considered geochemically inert. Our results demonstrate that Cr(III) within ultramafic- and serpentinite-derived soils/sediments can be oxidized and dissolved through natural processes, leading to hazardous levels of aqueous Cr(VI) in surface and groundwater. PMID- 17420455 TI - Insertion of group II intron retroelements after intrinsic transcriptional terminators. AB - Mobile DNAs use many mechanisms to minimize damage to their hosts. Here we show that a subclass of group II introns avoids host damage by inserting directly after transcriptional terminator motifs in bacterial genomes (stem-loops followed by Ts). This property contrasts with the site-specific behavior of most group II introns, which insert into homing site sequences. Reconstituted ribonucleo protein particles of the Bacillus halodurans intron B.h.I1 are shown to reverse splice into DNA targets in vitro but require the DNA to be single-stranded and fold into a stem-loop analogous to the RNA structure that forms during transcription termination. Recognition of this DNA stem-loop motif accounts for in vivo target specificity. Insertion after terminators is a previously unrecognized strategy for a selfish DNA because it prevents interruption of coding sequences and restricts expression of the mobile DNA after integration. PMID- 17420456 TI - High-affinity single-domain binding proteins with a binary-code interface. AB - High degrees of sequence and conformation complexity found in natural protein interaction interfaces are generally considered essential for achieving tight and specific interactions. However, it has been demonstrated that specific antibodies can be built by using an interface with a binary code consisting of only Tyr and Ser. This surprising result might be attributed to yet undefined properties of the antibody scaffold that uniquely enhance its capacity for target binding. In this work we tested the generality of the binary-code interface by engineering binding proteins based on a single-domain scaffold. We show that Tyr/Ser binary code interfaces consisting of only 15-20 positions within a fibronectin type III domain (FN3; 95 residues) are capable of producing specific binding proteins (termed "monobodies") with a low-nanomolar K(d). A 2.35-A x-ray crystal structure of a monobody in complex with its target, maltose-binding protein, and mutation analysis revealed dominant contributions of Tyr residues to binding as well as striking molecular mimicry of a maltose-binding protein substrate, beta cyclodextrin, by the Tyr/Ser binary interface. This work suggests that an interaction interface with low chemical diversity but with significant conformational diversity is generally sufficient for tight and specific molecular recognition, providing fundamental insights into factors governing protein protein interactions. PMID- 17420457 TI - Exceptional transmission of plastids and mitochondria from the transplastomic pollen parent and its impact on transgene containment. AB - Plastids in Nicotiana tabacum are normally transmitted to the progeny by the maternal parent only. However, low-frequency paternal plastid transmission has been reported in crosses involving parents with an alien cytoplasm. Our objective was to determine whether paternal plastids are transmitted in crosses between parents with the normal cytoplasm. The transplastomic father lines carried a spectinomycin resistance (aadA) transgene incorporated in the plastid genome. The mother lines in the crosses were either (i) alloplasmic, with the Nicotiana undulata cytoplasm that confers cytoplasmic male sterility (CMS92) or (ii) normal, with the fertile N. tabacum cytoplasm. Here we report that plastids from the transplastomic father were transmitted in both cases at low (10(-4)-10(-5)) frequencies; therefore, rare paternal pollen transmission is not simply due to breakdown of normal controls caused by the alien cytoplasm. Furthermore, we have found that the entire plastid genome was transmitted by pollen rather than small plastid genome (ptDNA) fragments. Interestingly, the plants, which inherited paternal plastids, also carried paternal mitochondrial DNA, indicating cotransmission of plastids and mitochondria in the same pollen. The detection of rare paternal plastid transmission described here was facilitated by direct selection for the transplastomic spectinomycin resistance marker in tissue culture; therefore, recovery of rare paternal plastids in the germline is less likely to occur under field conditions. PMID- 17420458 TI - Structural basis for reversible photobleaching of a green fluorescent protein homologue. AB - Fluorescent protein (FP) variants that can be reversibly converted between fluorescent and nonfluorescent states have proven to be a catalyst for innovation in the field of fluorescence microscopy. However, the structural basis of the process remains poorly understood. High-resolution structures of a FP derived from Clavularia in both the fluorescent and the light-induced nonfluorescent states reveal that the rapid and complete loss of fluorescence observed upon illumination with 450-nm light results from cis-trans isomerization of the chromophore. The photoinduced change in configuration from the well ordered cis isomer to the highly nonplanar and disordered trans isomer is accompanied by a dramatic rearrangement of internal side chains. Taken together, the structures provide an explanation for the loss of fluorescence upon illumination, the slow light-independent recovery, and the rapid light-induced recovery of fluorescence. The fundamental mechanism appears to be common to all of the photoactivatable and reversibly photoswitchable FPs reported to date. PMID- 17420459 TI - Determining the transgene containment level provided by chloroplast transformation. AB - Plastids (chloroplasts) are maternally inherited in most crops. Maternal inheritance excludes plastid genes and transgenes from pollen transmission. Therefore, plastid transformation is considered a superb tool for ensuring transgene containment and improving the biosafety of transgenic plants. Here, we have assessed the strictness of maternal inheritance and the extent to which plastid transformation technology confers an increase in transgene confinement. We describe an experimental system facilitating stringent selection for occasional paternal plastid transmission. In a large screen, we detected low level paternal inheritance of transgenic plastids in tobacco. Whereas the frequency of transmission into the cotyledons of F(1) seedlings was approximately 1.58 x 10(-5) (on 100% cross-fertilization), transmission into the shoot apical meristem was significantly lower (2.86 x 10(-6)). Our data demonstrate that plastid transformation provides an effective tool to increase the biosafety of transgenic plants. However, in cases where pollen transmission must be prevented altogether, stacking with other containment methods will be necessary to eliminate the residual outcrossing risk. PMID- 17420460 TI - Fronts and pulses in an enzymatic reaction catalyzed by glucose oxidase. AB - Waves and patterns in living systems are often driven by biochemical reactions with enzymes as catalysts and regulators. We present a reaction-diffusion system catalyzed by the enzyme glucose oxidase that exhibits traveling wave patterns in a spatially extended medium. Fronts and pulses propagate as a result of the coupling between the enzyme-catalyzed autocatalytic production and diffusion of hydrogen ions. A mathematical model qualitatively explains the experimental observations. PMID- 17420461 TI - Geochemical proxies of North American freshwater routing during the Younger Dryas cold event. AB - The Younger Dryas cold interval represents a time when much of the Northern Hemisphere cooled from approximately 12.9 to 11.5 kiloyears B.P. The cause of this event, which has long been viewed as the canonical example of abrupt climate change, was initially attributed to the routing of freshwater to the St. Lawrence River with an attendant reduction in Atlantic meridional overturning circulation. However, this mechanism has recently been questioned because current proxies and dating techniques have been unable to confirm that eastward routing with an increase in freshwater flux occurred during the Younger Dryas. Here we use new geochemical proxies (DeltaMg/Ca, U/Ca, and (87)Sr/(86)Sr) measured in planktonic foraminifera at the mouth of the St. Lawrence estuary as tracers of freshwater sources to further evaluate this question. Our proxies, combined with planktonic delta(18)O(seawater) and delta(13)C, confirm that routing of runoff from western Canada to the St. Lawrence River occurred at the start of the Younger Dryas, with an attendant increase in freshwater flux of 0.06 +/- 0.02 Sverdrup (1 Sverdrup = 10(6) m(3).s(-1)). This base discharge increase is sufficient to have reduced Atlantic meridional overturning circulation and caused the Younger Dryas cold interval. In addition, our data indicate subsequent fluctuations in the freshwater flux to the St. Lawrence River of approximately 0.06-0.12 Sverdrup, thus explaining the variability in the overturning circulation and climate during the Younger Dryas. PMID- 17420463 TI - Combined climate and carbon-cycle effects of large-scale deforestation. AB - The prevention of deforestation and promotion of afforestation have often been cited as strategies to slow global warming. Deforestation releases CO(2) to the atmosphere, which exerts a warming influence on Earth's climate. However, biophysical effects of deforestation, which include changes in land surface albedo, evapotranspiration, and cloud cover also affect climate. Here we present results from several large-scale deforestation experiments performed with a three dimensional coupled global carbon-cycle and climate model. These simulations were performed by using a fully three-dimensional model representing physical and biogeochemical interactions among land, atmosphere, and ocean. We find that global-scale deforestation has a net cooling influence on Earth's climate, because the warming carbon-cycle effects of deforestation are overwhelmed by the net cooling associated with changes in albedo and evapotranspiration. Latitude specific deforestation experiments indicate that afforestation projects in the tropics would be clearly beneficial in mitigating global-scale warming, but would be counterproductive if implemented at high latitudes and would offer only marginal benefits in temperate regions. Although these results question the efficacy of mid- and high-latitude afforestation projects for climate mitigation, forests remain environmentally valuable resources for many reasons unrelated to climate. PMID- 17420462 TI - Unrestrained erythroblast development in Nix-/- mice reveals a mechanism for apoptotic modulation of erythropoiesis. AB - Normal production of RBCs requires that the antiapoptotic protein Bcl-xl be induced at end stages of differentiation in response to erythropoietin (Epo) signaling. The critical proapoptotic pathways inhibited by Bcl-xl in erythroblasts are unknown. We used gene targeting in the mouse to evaluate the BH3-only factor Nix, which is transcriptionally up-regulated during Epo stimulated in vitro erythrocyte differentiation. Nix null mice are viable and fertile. Peripheral blood counts revealed a profound reticulocytosis and thrombocytosis despite normal serum Epo levels and blood oxygen tension. Nix null mice exhibited massive splenomegaly, with splenic and bone marrow erythroblastosis and reduced apoptosis in vivo during erythrocyte maturation. Hematopoietic progenitor populations were unaffected. Cultured Nix null erythroid cells were hypersensitive to Epo and resistant to apoptosis stimulated by cytokine deprivation and calcium ionophore. Transcriptional profiling of Nix null spleens revealed increased expression of cell cycle and erythroid genes, including Bcl-xl, and diminished expression of cell death and B cell-related genes. Thus, cell-autonomous Nix-mediated apoptosis in opposition to the Epo induced erythroblast survival pathway appears indispensable for regulation of erythrocyte production and maintenance of hematological homeostasis. These results suggest that physiological codependence and coordinated regulation of pro and antiapoptotic Bcl2 family members may represent a general regulatory paradigm in hematopoiesis. PMID- 17420464 TI - Noisy information processing through transcriptional regulation. AB - Cells must respond to environmental changes to remain viable, yet the information they receive is often noisy. Through a biochemical implementation of Bayes's rule, we show that genetic networks can act as inference modules, inferring from intracellular conditions the likely state of the extracellular environment and regulating gene expression appropriately. By considering a two-state environment, either poor or rich in nutrients, we show that promoter occupancy is proportional to the (posterior) probability of the high nutrient state given current intracellular information. We demonstrate that single-gene networks inferring and responding to a high environmental state infer best when negatively controlled, and those inferring and responding to a low environmental state infer best when positively controlled. Our interpretation is supported by experimental data from the lac operon and should provide a basis for both understanding more complex cellular decision-making and designing synthetic inference circuits. PMID- 17420465 TI - A missense mutation in the 3-ketodihydrosphingosine reductase FVT1 as candidate causal mutation for bovine spinal muscular atrophy. AB - The bovine form of the autosomal recessive neurodegenerative disease spinal muscular atrophy (SMA) shows striking similarity to the human form of the disease. It has, however, been mapped to a genomic region not harboring the bovine orthologue of the SMN gene, mutation of which causes human SMA. After refinement of the mapping results we analyzed positional and functional candidate genes. One of three candidate genes, FVT1, encoding 3-ketodihydrosphingosine reductase, which catalyzes a crucial step in the glycosphingolipid metabolism, showed a G-to-A missense mutation that changes Ala-175 to Thr. The identified mutation is limited to SMA-affected animals and carriers and always appears in context of the founder haplotype. The Ala variant found in healthy animals showed the expected 3-ketodihydrosphingosine reductase activity in an in vitro enzyme assay. Importantly, the Thr variant found in SMA animals showed no detectable activity. Surprisingly, in an in vivo assay the mutated gene complements the growth defect of a homologous yeast knockout strain as well as the healthy variant. This finding explains the viability of affected newborn calves and the later neuron-specific onset of the disease, which might be due to the high sensitivity of these neurons to changes in housekeeping functions. Taken together, the described mutation in FVT1 is a strong candidate for causality of SMA in cattle. This result provides an animal model for understanding the underlying mechanisms of the development of SMA and will allow efficient selection against the disease in cattle. PMID- 17420466 TI - Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans. AB - In the cilia of the nematode Caenorhabditis elegans, anterograde intraflagellar transport (IFT) is mediated by two kinesin-2 complexes, kinesin II and OSM-3 kinesin. These complexes function together in the cilia middle segments, whereas OSM-3 alone mediates transport in the distal segments. Not much is known about the mechanisms that compartmentalize the kinesin-2 complexes or how transport by both kinesins is coordinated. Here, we identify DYF-5, a conserved MAP kinase that plays a role in these processes. Fluorescence microscopy and EM revealed that the cilia of dyf-5 loss-of-function (lf) animals are elongated and are not properly aligned into the amphid channel. Some cilia do enter the amphid channel, but the distal ends of these cilia show accumulation of proteins. Consistent with these observations, we found that six IFT proteins accumulate in the cilia of dyf 5(lf) mutants. In addition, using genetic analyses and live imaging to measure the motility of IFT proteins, we show that dyf-5 is required to restrict kinesin II to the cilia middle segments. Finally, we show that, in dyf-5(lf) mutants, OSM 3 moves at a reduced speed and is not attached to IFT particles. We propose that DYF-5 plays a role in the undocking of kinesin II from IFT particles and in the docking of OSM-3 onto IFT particles. PMID- 17420467 TI - Asymmetric binding of transferrin receptor to parvovirus capsids. AB - Although many viruses are icosahedral when they initially bind to one or more receptor molecules on the cell surface, such an interaction is asymmetric, probably causing a breakdown in the symmetry and conformation of the original infecting virion in preparation for membrane penetration and release of the viral genome. Cryoelectron microscopy and biochemical analyses show that transferrin receptor, the cellular receptor for canine parvovirus, can bind to only one or a few of the 60 icosahedrally equivalent sites on the virion, indicating that either canine parvovirus has inherent asymmetry or binding of receptor induces asymmetry. The asymmetry of receptor binding to canine parvovirus is reminiscent of the special portal in tailed bacteriophages and some large, icosahedral viruses. Asymmetric interactions of icosahedral viruses with their hosts might be a more common phenomenon than previously thought and may have been obscured by averaging in previous crystallographic and electron microscopic structure determinations. PMID- 17420468 TI - Lung metastasis genes couple breast tumor size and metastatic spread. AB - The association between large tumor size and metastatic risk in a majority of clinical cancers has led to questions as to whether these observations are causally related or whether one is simply a marker for the other. This is partly due to an uncertainty about how metastasis-promoting gene expression changes can arise in primary tumors. We investigated this question through the analysis of a previously defined "lung metastasis gene-expression signature" (LMS) that mediates experimental breast cancer metastasis selectively to the lung and is expressed by primary human breast cancer with a high risk for developing lung metastasis. Experimentally, we demonstrate that the LMS promotes primary tumor growth that enriches for LMS(+) cells, and it allows for intravasation after reaching a critical tumor size. Clinically, this corresponds to LMS(+) tumors being larger at diagnosis compared with LMS(-) tumors and to a marked rise in the incidence of metastasis after LMS(+) tumors reach 2 cm. Patients with LMS expressing primary tumors selectively fail in the lung compared with the bone or other visceral sites and have a worse overall survival. The mechanistic linkage between metastasis gene expression, accelerated tumor growth, and likelihood of metastatic recurrence provided by the LMS may help to explain observations of prognostic gene signatures in primary cancer and how tumor growth can both lead to metastasis and be a marker for cells destined to metastasize. PMID- 17420469 TI - Linking crenarchaeal and bacterial nitrification to anammox in the Black Sea. AB - Active expression of putative ammonia monooxygenase gene subunit A (amoA) of marine group I Crenarchaeota has been detected in the Black Sea water column. It reached its maximum, as quantified by reverse-transcription quantitative PCR, exactly at the nitrate maximum or the nitrification zone modeled in the lower oxic zone. Crenarchaeal amoA expression could explain 74.5% of the nitrite variations in the lower oxic zone. In comparison, amoA expression by gamma proteobacterial ammonia-oxidizing bacteria (AOB) showed two distinct maxima, one in the modeled nitrification zone and one in the suboxic zone. Neither the amoA expression by crenarchaea nor that by beta-proteobacterial AOB was significantly elevated in this latter zone. Nitrification in the suboxic zone, most likely microaerobic in nature, was verified by (15)NO(2)(-) and (15)N(15)N production in (15)NH(4)(+) incubations with no measurable oxygen. It provided a direct local source of nitrite for anammox in the suboxic zone. Both ammonia-oxidizing crenarchaea and gamma-proteobacterial AOB were important nitrifiers in the Black Sea and were likely coupled to anammox in indirect and direct manners respectively. Each process supplied about half of the nitrite required by anammox, based on (15)N-incubation experiments and modeled calculations. Because anammox is a major nitrogen loss in marine suboxic waters, such nitrification anammox coupling potentially occurring also in oceanic oxygen minimum zones would act as a short circuit connecting regenerated ammonium to direct nitrogen loss, thus reducing the presumed direct contribution from deep-sea nitrate. PMID- 17420470 TI - SGT1 is essential for Nod1 activation. AB - The Nod-like receptor family in man contains proteins that recognize invasive bacteria. Nod1, a member of this family, is activated by specific peptidoglycan derived muropeptides that contain meso-diaminopimelic acid. Plants contain a large family of proteins known as resistance (R) proteins that have common structural features with the Nod-like receptors and are essential for protection against a variety of plant pathogens. Extensive genetic studies have shown that the R protein function is determined by multiple proteins including SGT1, Rar1, and HSP90. Here we show that SGT1 positively regulates Nod1 activation. Depletion of SGT1 with siRNA did not affect stability of Nod1 protein or of downstream signaling molecules but did prevent multiple cellular responses associated with Nod1 activation. In contrast, depletion of the mammalian orthologue of Rar1, Chp1, had no effect on Nod1-dependent cellular activation. Finally, depletion of HSP90 or addition of a pharmacologic inhibitor of HSP90 resulted in loss of Nod1 protein. Thus, we show common regulatory pathways in plant R protein and human Nod1-dependent pathways and provide the basis for understanding the Nod1 pathway. PMID- 17420471 TI - A mechanism for transcriptional repression dependent on the BRCA1 E3 ubiquitin ligase. AB - Loss of function of the tumor suppressor protein BRCA1 is responsible for a high percentage of familial and also sporadic breast cancers. Early work identified a stimulatory transcriptional coactivator function for the BRCA1 protein, and more recently, BRCA1 has been implicated in transcriptional repression, although few examples of repressed genes have been characterized. We recently used an in vitro transcription assay to identify a biochemical mechanism that explained the BRCA1 stimulatory activity. In this study, we identified an ubiquitin-dependent mechanism by which BRCA1 inhibits transcription. BRCA1 ubiquitinates the transcriptional preinitiation complex, preventing stable association of TFIIE and TFIIH, and thus blocks the initiation of mRNA synthesis. What is striking about this mechanism of regulation by BRCA1 is that the ubiquitination of the preinitiation complex is not targeting proteins for degradation by the proteasome, nor are ubiquitin receptors modifying the activity, but rather the ubiquitin moiety itself interferes with the assembly of basal transcription factors at the promoter. Using RNAi to knockdown expression of the endogenous BRCA1 protein, we assessed the level of repression dependent on BRCA1 in the cell, and we found that BRCA1 is at least as significant a transcriptional repressor as it is an activator. These results define a biochemical mechanism by which the BRCA1 enzymatic activity regulates a key cellular process. PMID- 17420472 TI - A cationic host displaying positive cooperativity in water. AB - The affinity of guanidinium and Cu(II) containing hosts for polycarboxylate guests is studied in water by using UV-visible spectroscopy and isothermal titration calorimetry. By combining a Cu(II) coordination site and a diguanidinium moiety around a single scaffold it is found that the Gibbs free energy release upon binding is greater than could be expected based on the sum of the free energy released by the binding functional groups when operating in isolation. This effect is known as positive cooperativity and has rarely been observed with synthetic hosts in water. The isothermal titration calorimetry data suggest that the source of this positive cooperativity is enthalpic in origin. We propose that covalently positioning the cationic binding moieties in close proximity destabilizes the unbound host, and thereby the stabilizing effect of binding the anionic carboxylates is exothermic and also, the source of positive cooperativity observed herein. PMID- 17420473 TI - Inducible overexpression of wild-type prion protein in the muscles leads to a primary myopathy in transgenic mice. AB - The prion protein (PrP) level in muscle has been reported to be elevated in patients with inclusion-body myositis, polymyositis, dermatomyositis, and neurogenic muscle atrophy, but it is not clear whether the elevated PrP accumulation in the muscles is sufficient to cause muscle diseases. We have generated transgenic mice with muscle-specific expression of PrP under extremely tight regulation by doxycycline, and we have demonstrated that doxycycline induced overexpression of PrP strictly limited to muscles leads to a myopathy characterized by increased variation of myofiber size, centrally located nuclei, and endomysial fibrosis, in the absence of intracytoplasmic inclusions, rimmed vacuoles, or any evidence of a neurogenic disorder. The PrP-induced myopathy correlates with accumulation of an N-terminal truncated PrP fragment in the muscle, and the muscular PrP displayed consistent mild resistance to protease digestion. Our findings indicate that overexpression of wild-type PrP in skeletal muscles is sufficient to cause a primary myopathy with no signs of peripheral neuropathy, possibly due to accumulation of a cytotoxic truncated form of PrP and/or PrP aggregation. PMID- 17420474 TI - Combinatorial RNAi for quantitative protein network analysis. AB - The elucidation of cross-talk events between intersecting signaling pathways is one main challenge in biological research. The complexity of protein networks, composed of different pathways, requires novel strategies and techniques to reveal relevant interrelations. Here, we established a combinatorial RNAi strategy for systematic single, double, and triple knockdown, and we measured the residual mRNAs and proteins quantitatively by quantitative real-time PCR and reverse-phase protein arrays, respectively, as a prerequisite for data analysis. Our results show that the parallel knockdown of at least three different genes is feasible while keeping both untargeted silencing and cytotoxicity low. The technique was validated by investigating the interplay of tyrosine kinase receptor ErbB2 and its downstream targets Akt-1 and MEK1 in cell invasion. This experimental approach combines multiple gene knockdown with a subsequent quantitative validation of reduced protein expression and is a major advancement toward the analysis of signaling pathways in systems biology. PMID- 17420475 TI - Enhanced EGFP-chromophore-assisted laser inactivation using deficient cells rescued with functional EGFP-fusion proteins. AB - Chromophore-assisted laser inactivation (CALI) is a light-mediated technique that offers precise spatiotemporal control of protein inactivation, enabling better understanding of the protein's role in cell function. EGFP has been used effectively as a CALI chromophore, and its cotranslational attachment to the target protein avoids having to use exogenously added labeling reagents. A potential drawback to EGFP-CALI is that the CALI phenotype can be obscured by the endogenous, unlabeled protein that is not susceptible to light inactivation. Performing EGFP-CALI experiments in deficient cells rescued with functional EGFP fusion proteins permits more complete loss of function to be achieved. Here, we present a modified lentiviral system for rapid and efficient generation of knockdown cell lines complemented with physiological levels of EGFP-fusion proteins. We demonstrate that CALI of EGFP-CapZbeta increases uncapped actin filaments, resulting in enhanced filament growth and the formation of numerous protrusive structures. We show that these effects are completely dependent upon knocking down the endogenous protein. We also demonstrate that CALI of EGFP-Mena in Mena/VASP-deficient cells stabilizes lamellipodial protrusions. PMID- 17420476 TI - Algorithm for model validation: theory and applications. AB - Validation is often defined as the process of determining the degree to which a model is an accurate representation of the real world from the perspective of its intended uses. Validation is crucial as industries and governments depend increasingly on predictions by computer models to justify their decisions. We propose to formulate the validation of a given model as an iterative construction process that mimics the often implicit process occurring in the minds of scientists. We offer a formal representation of the progressive build-up of trust in the model. Thus, we replace static claims on the impossibility of validating a given model by a dynamic process of constructive approximation. This approach is better adapted to the fuzzy, coarse-grained nature of validation. Our procedure factors in the degree of redundancy versus novelty of the experiments used for validation as well as the degree to which the model predicts the observations. We illustrate the methodology first with the maturation of quantum mechanics as the arguably best established physics theory and then with several concrete examples drawn from some of our primary scientific interests: a cellular automaton model for earthquakes, a multifractal random walk model for financial time series, an anomalous diffusion model for solar radiation transport in the cloudy atmosphere, and a computational fluid dynamics code for the Richtmyer-Meshkov instability. PMID- 17420477 TI - Quantifying the genetic influence on mammalian vascular tree structure. AB - The ubiquity of fractal vascular trees throughout the plant and animal kingdoms is postulated to be due to evolutionary advantages conferred through efficient distribution of nutrients to multicellular organisms. The implicit, and untested, assertion in this theory is that the geometry of vascular trees is heritable. Because vascular trees are constructed through the iterative use of signaling pathways modified by local factors at each step of the branching process, we sought to investigate how genetic and nongenetic influences are balanced to create vascular trees and the regional distribution of nutrients through them. We studied the spatial distribution of organ blood flow in armadillos because they have genetically identical littermates, allowing us to quantify the genetic influence. We determined that the regional distribution of blood flow is strongly correlated between littermates (r(2) = 0.56) and less correlated between unrelated animals (r(2) = 0.36). Using an ANOVA model, we estimate that 67% of the regional variability in organ blood flow is genetically controlled. We also used fractal analysis to characterize the distribution of organ blood flow and found shared patterns within the lungs and hearts of related animals, suggesting common control over the vascular development of these two organs. We conclude that the geometries of fractal vascular trees are heritable and could be selected through evolutionary pressures. Furthermore, considerable postgenetic modifications may allow vascular trees to adapt to local factors and provide a flexibility that would not be possible in a rigid system. PMID- 17420478 TI - A cortical potential reflecting cardiac function. AB - Emotional trauma and psychological stress can precipitate cardiac arrhythmia and sudden death through arrhythmogenic effects of efferent sympathetic drive. Patients with preexisting heart disease are particularly at risk. Moreover, generation of proarrhythmic activity patterns within cerebral autonomic centers may be amplified by afferent feedback from a dysfunctional myocardium. An electrocortical potential reflecting afferent cardiac information has been described, reflecting individual differences in interoceptive sensitivity (awareness of one's own heartbeats). To inform our understanding of mechanisms underlying arrhythmogenesis, we extended this approach, identifying electrocortical potentials corresponding to the cortical expression of afferent information about the integrity of myocardial function during stress. We measured changes in cardiac response simultaneously with electroencephalography in patients with established ventricular dysfunction. Experimentally induced mental stress enhanced cardiovascular indices of sympathetic activity (systolic blood pressure, heart rate, ventricular ejection fraction, and skin conductance) across all patients. However, the functional response of the myocardium varied; some patients increased, whereas others decreased, cardiac output during stress. Across patients, heartbeat-evoked potential amplitude at left temporal and lateral frontal electrode locations correlated with stress-induced changes in cardiac output, consistent with an afferent cortical representation of myocardial function during stress. Moreover, the amplitude of the heartbeat-evoked potential in the left temporal region reflected the proarrhythmic status of the heart (inhomogeneity of left ventricular repolarization). These observations delineate a cortical representation of cardiac function predictive of proarrhythmic abnormalities in cardiac repolarization. Our findings highlight the dynamic interaction of heart and brain in stress-induced cardiovascular morbidity. PMID- 17420479 TI - SLP-76 mediates and maintains activation of the Tec family kinase ITK via the T cell antigen receptor-induced association between SLP-76 and ITK. AB - ITK (IL-2-inducible T cell kinase), a Tec family protein tyrosine kinase (PTK), is one of three PTKs required for T cell antigen receptor (TCR)-induced activation of phospholipase C-gamma1 (PLC-gamma1). Like Src and Abl family PTKs, ITK adopts an inactive, "closed" conformation, and its conversion to the active conformation is not well understood, nor have its direct substrates been identified. In a side-by-side comparison of ITK and ZAP-70 (zeta chain-associated protein kinase of 70 kDa), ITK efficiently phosphorylated Y(783) and Y(775) of PLC-gamma1, two phosphorylation sites that are critical for its activation, whereas ZAP-70 did not. SLP-76 (SH2-domain-containing leukocyte protein of 76 kDa), an adaptor required for TCR-induced activation of PLC-gamma1, was required for the phosphorylation of both PLC-gamma1 sites in intact cells. Furthermore, this event depended on the N-terminal tyrosines of SLP-76. Likewise, SLP-76, particularly its N-terminal tyrosines, was required for TCR-induced tyrosine phosphorylation and activation of ITK but was not required for the phosphorylation or activation of ZAP-70. Both ZAP-70 and ITK phosphorylated SLP 76 in vitro; thus, both PTKs are potential regulators of SLP-76, but only ITK is regulated by SLP-76. Upon TCR stimulation, a small fraction of ITK bound to SLP 76. This fraction, however, encompassed most of the catalytically active ITK. Catalytic activity was lost upon mild elution of ITK from the SLP-76-nucleated complex but was restored upon reconstitution of the complex. We propose that SLP 76 is required for ITK activation; furthermore, an ongoing physical interaction between SLP-76 and ITK is required to maintain ITK in an active conformation. PMID- 17420480 TI - Omics-based identification of Arabidopsis Myb transcription factors regulating aliphatic glucosinolate biosynthesis. AB - Understanding plant metabolism as an integrated system is essential for metabolic engineering aimed at the effective production of compounds useful to human life and the global environment. The "omics" approach integrates transcriptome and metabolome data into a single data set and can lead to the identification of unknown genes and their regulatory networks involved in metabolic pathways of interest. One of the intriguing, although poorly described metabolic pathways in plants is the biosynthesis of glucosinolates (GSLs), a group of bioactive secondary products derived from amino acids that are found in the family Brassicaceae. Here we report the discovery of two R2R3-Myb transcription factors that positively control the biosynthesis of GSLs in Arabidopsis thaliana by an integrated omics approach. Combined transcriptome coexpression analysis of publicly available, condition-independent data and the condition-specific (i.e., sulfur-deficiency) data identified Myb28 and Myb29 as candidate transcription factor genes specifically involved in the regulation of aliphatic GSL production. Analysis of a knockout mutant and ectopic expression of the gene demonstrated that Myb28 is a positive regulator for basal-level production of aliphatic GSLs. Myb29 presumably plays an accessory function for methyl jasmonate-mediated induction of a set of aliphatic GSL biosynthetic genes. Overexpression of Myb28 in Arabidopsis-cultured suspension cells, which do not normally synthesize GSLs, resulted in the production of large amounts of GSLs, suggesting the possibility of efficient industrial production of GSLs by manipulation of these transcription factors. A working model for regulation of GSL production involving these genes, renamed Production of Methionine-Derived Glucosinolate (PMG) 1 and 2, are postulated. PMID- 17420482 TI - Managing physician shortages: we are not doing enough. PMID- 17420481 TI - 2006 Canadian clinical practice guidelines on the management and prevention of obesity in adults and children [summary]. PMID- 17420483 TI - Estimated 1500 Canadians studying medicine abroad. PMID- 17420484 TI - New deputy editor dedicated to knowledge transfer. PMID- 17420485 TI - Saskatchewan successfully implementing best practices. PMID- 17420486 TI - Proposed Alberta physician agreement sets new template. PMID- 17420487 TI - Pharmacare for children slow to gain ground in NS. PMID- 17420488 TI - Management of hypothyroidism during pregnancy. PMID- 17420489 TI - Upper-extremity deep-vein thrombosis in an elderly man. PMID- 17420490 TI - The Canadian contribution to the US physician workforce. AB - BACKGROUND: A physician shortage has been declared in both Canada and the United States. We sought to examine the migration pattern of Canadian-trained physicians to the United States, the contribution of this migration to the Canadian physician shortage and policy options in light of competing shortages in both countries. METHODS: We performed a cross-sectional analysis of the 2004 and 2006 American Medical Association Physician Masterfiles, the 2002 Area Resource File and data from the Canadian Institute for Health Information, the Canadian Medical Association and the Association of Faculties of Medicine of Canada. We describe the migration pattern of Canadian medical school graduates to the United States, the number of Canadian-trained physicians in the United States in 2006, the proportion who were in active practice, the proportion who were practising in rural or underserved areas and the annual contribution of Canadian-trained physicians to the US physician workforce. RESULTS: Two-thirds of the 12 040 Canadian-educated physicians living in the United States in 2006 were practising in direct patient care, 1023 in rural areas. About 186, or 1 in 9, Canadian educated physicians from each graduating class joined the US physician workforce providing direct patient care. Canadian-educated physicians are more likely than US-educated physicians to practise in rural areas. INTERPRETATION: Minimizing emigration, and perhaps recruiting physicians to return to Canada, could reduce physician shortages, particularly in subspecialties and rural areas. In light of competing physician shortages, it will be important to consider policy options that reduce emigration, improve access to care and reduce reliance on physicians from developing countries. PMID- 17420491 TI - The appropriateness of asymmetry tests for publication bias in meta-analyses: a large survey. AB - BACKGROUND: Statistical tests for funnel-plot asymmetry are common in meta analyses. Inappropriate application can generate misleading inferences about publication bias. We aimed to measure, in a survey of meta-analyses, how frequently the application of these tests would be not meaningful or inappropriate. METHODS: We evaluated all meta-analyses of binary outcomes with e 3 studies in the Cochrane Database of Systematic Reviews (2003, issue 2). A separate, restricted analysis was confined to the largest meta-analysis in each of the review articles. In each meta-analysis, we assessed whether criteria to apply asymmetry tests were met: no significant heterogeneity, I2 < 50%, e 10 studies (with statistically significant results in at least 1) and ratio of the maximal to minimal variance across studies > 4. We performed a correlation and 2 regression asymmetry tests and evaluated their concordance. Finally, we sampled 60 meta-analyses from print journals in 2005 that cited use of the standard regression test. RESULTS: A total of 366 of 6873 (5%) and 98 of 846 meta-analyses (12%) in the wider and restricted Cochrane data set, respectively, would have qualified for use of asymmetry tests. Asymmetry test results were significant in 7%-18% of the meta-analyses. Concordance between the 3 tests was modest (estimated k 0.33-0.66). Of the 60 journal meta-analyses, 7 (12%) would qualify for asymmetry tests; all 11 claims for identification of publication bias were made in the face of large and significant heterogeneity. INTERPRETATION: Statistical conditions for employing asymmetry tests for publication bias are absent from most meta-analyses; yet, in medical journals these tests are performed often and interpreted erroneously. PMID- 17420492 TI - Falsely elevated point-of-care lactate measurement after ingestion of ethylene glycol. AB - BACKGROUND: A patient presented with severe acidosis and a point-of-care lactate measurement of 42 mmol/L. Mesenteric ischemia was suspected, with a potential need for laparotomy; however, plasma lactate measurements were below 4 mmol/L. Ethylene glycol ingestion was subsequently diagnosed. We therefore wished to determine why discrepancies in lactate measurements occur and whether this "lactate gap" could be clinically useful. METHODS: We phlebotomized blood, added various concentrations of metabolites of ethylene glycol, and tested the resulting samples with the 5 most common lactate analyzers. RESULTS: With the Radiometer 700 point-of-care analyzer, glycolate addition resulted in an artifactual, massive lactate elevation, even at low glycolate concentrations. Another major ethylene glycol metabolite, glyoxylate (but not oxalate or formate), caused similar elevations. The i-STAT and Bayer point-of-care analyzers and the Beckman and Vitros laboratory analyzers reported minimal lactate elevations. Lactate gap was determined by comparing the Radiometer result with the corresponding result from any of the other analyzers. INTERPRETATION: We demonstrated how inappropriate laparotomy or delayed therapy might occur if clinicians are unaware of this phenomenon or have access to only a single analyzer. We also showed that lactate gap can be exploited to expedite treatment, diagnose late ethylene-glycol ingestion and terminate dialysis. By comparing lactate results from the iSTAT or Bayer devices with that from the Radiometer, ethylene-glycol ingestion can be diagnosed at the point of care. This can expedite diagnosis and treatment by hours, compared with waiting for laboratory results for plasma ethylene glycol. PMID- 17420493 TI - Synopsis of the 2006 Canadian clinical practice guidelines on the management and prevention of obesity in adults and children. PMID- 17420494 TI - Management and prevention of obesity in adults and children. PMID- 17420495 TI - Hypertriglyceridemia: its etiology, effects and treatment. AB - Elevated plasma triglyceride concentration is a common biochemical finding, but the evidence for the benefit of treating this lipid disturbance remains less robust than that for treating elevated low-density lipoprotein-cholesterol. Part of the difficulty in the provision of specific recommendations has been the frequent coexistence of elevated triglycerides with other conditions that affect cardiovascular disease risk, such as depressed high-density lipoprotein cholesterol, obesity, metabolic syndrome, proinflammatory and prothrombotic biomarkers, and type 2 diabetes. Recent investigations of outcomes of cardiovascular disease when medications are used to reduce triglyceride levels suggest that, although a net benefit probably exists, both relative and absolute risk reductions seem underwhelming when compared with the benefit of reducing low density lipoprotein-cholesterol levels with treatment. However, the totality of evidence suggests that elevated triglyceride levels likely contribute independently to increased risk of cardiovascular disease, although there is no consensus about appropriate target levels. Furthermore, severe hypertriglyceridemia is associated with an increased risk of acute pancreatitis, irrespective of its effect on risk of cardiovascular disease. We review the causes and classification of elevated triglyceride levels, the clinical manifestations of primary hypertriglyceridemia and the management of patients with elevated triglyceride levels. PMID- 17420496 TI - Improving organ donation in Canada. PMID- 17420498 TI - Bias in revascularization study. PMID- 17420499 TI - Improving organ donation in Canada. PMID- 17420500 TI - Improving organ donation in Canada. PMID- 17420502 TI - Auspitz sign-off. PMID- 17420504 TI - Treatment of chronic respiratory diseases in obese people. PMID- 17420505 TI - Uninsured patients undergoing dialysis in Greece. PMID- 17420506 TI - Humeral avulsion of the glenohumeral ligaments: the HAGL lesion. AB - BACKGROUND: Anterior instability of the shoulder is a frequently encountered clinical problem that is seen after avulsion of the capsulolabral complex from the glenoid rim (Bankart lesion). Recently, it has been determined that avulsion of the glenohumeral ligaments is an infrequent but important cause of recurrent instability after shoulder injury. Although the various forms of this injury have been described in the literature, no standardized nomenclature exists. This is of concern given the need for different surgical approaches to repair the various forms of this injury and therefore the inherent need to accurately convey the location of the avulsion and presence of concomitant injuries. HYPOTHESIS: Based on the available literature for humeral avulsion of the glenohumeral ligament lesions, a nomenclature can be created to enhance the understanding and improve communication about these uncommon but important pathologic changes that occur with shoulder instability. STUDY DESIGN: Systematic review. METHODS: The findings pertaining to 6 patients with humeral avulsion of the glenohumeral ligament lesions encountered at our institution were combined with a literature search of the MEDLINE database conducted using the PubMed search engine of the National Library of Medicine and National Center for Biotechnology Information. RESULTS: The West Point nomenclature was developed to clearly describe the various forms of humeral avulsion of the gleno-humeral ligament lesions encountered as well as their associated injuries. CONCLUSIONS: The West Point nomenclature provides a practical and easy to understand means by which to classify humeral avulsion of the glenohumeral ligament lesions. This allows for more effective communication, which should result in improved clinical care. PMID- 17420507 TI - Differences in 3-dimensional shoulder kinematics between persons with multidirectional instability and asymptomatic controls. AB - BACKGROUND: Evidence that persons with multidirectional instability (MDI) of the shoulder have abnormal shoulder kinematics is limited. A kinematic description of scapulothoracic and glenohumeral motion can assist both conservative and surgical rehabilitative programs. HYPOTHESIS: Persons with MDI of the shoulder demonstrate increased anterior and inferior glenohumeral translation and decreased scapular upward rotation and increased scapular internal rotation compared with age matched and gender-matched asymptomatic controls. STUDY DESIGN: Controlled laboratory study. METHODS: Sixty-two subjects were recruited from an outpatient orthopaedic clinic. Subjects with MDI were matched according to age, gender, and hand dominance to asymptomatic controls. An electromagnetic motion capture system evaluated the 3-dimensional position of the trunk, scapula, and humerus during frontal and scapular plane elevation. A repeated measures analysis of variance evaluated joint positions and glenohumeral translations during 4 phases of elevation (0 degrees-30 degrees, 31 degrees-60 degrees, 61 degrees-90 degrees, and 91 degrees-120 degrees). RESULTS: When averaged across the 4 phases of elevation, persons with MDI demonstrated a significant decrease in scapular upward rotation in scapular plane abduction (8 degrees, P = .006) and abduction (5.8 degrees, P = .016) and increased internal rotation during scapular plane abduction (12.2 degrees, P = .03). Alterations in glenohumeral translations in the MDI group did not reach statistical significance (P = .54-.71). CONCLUSION: Abnormal scapular kinematics are seen in the MDI shoulder, highlighting the importance of incorporating scapular positioning and stability exercises during rehabilitation. Additional study is warranted concerning the efficacy of various rehabilitation programs, and also both surgical and nonsurgical interventions in this population. PMID- 17420508 TI - Arthroscopic management of femoroacetabular impingement: osteoplasty technique and literature review. AB - Morphological and spatial abnormalities of the proximal femur and acetabulum have been recently recognized as causes of femoroacetabular impingement. During joint motion in hips with femoroacetabular impingement, abnormal bony contact occurs, and soft tissue structures (chondral and labral) often fail. Femoroacetabular impingement has been reported to be a contributor to early-onset joint degeneration. Ganz et al have described good midterm success with an open surgical dislocation approach to reconstruct normal joint clearance. The purpose of this report is to discuss relevant literature and describe an arthroscopic approach to treat femoroacetabular impingement. This approach has particular relevance in high-demand patients, particularly in athletes seeking to return to high-level sport. PMID- 17420509 TI - Comparison of bioabsorbable pins and nails in the fixation of adult osteochondritis dissecans fragments of the knee: an outcome of 30 knees. AB - BACKGROUND: The optimal device for the fixation of osteochondritis dissecans fragments of the knee remains controversial and lacks long-term results. PURPOSE: To review a group of young adults with osteochondritis dissecans of the knee treated with arthroscopic fixation of the fragment using bioabsorbable pins and nails and to examine the medium-term outcome of the fixation via magnetic resonance imaging and clinical evaluation. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: Twenty-eight patients (30 knees) with osteochondritis dissecans of the knee were treated with arthroscopic fixation using bioabsorbable, self-reinforced poly-L-lactide pins and nails. All patients were young adult males with closed physes. The average follow-up time was 5.4 years (range, 3-12). At follow-up, magnetic resonance imaging studies were used to evaluate subchondral bone healing, and the outcome was evaluated by the Kujala score. RESULTS: The functional results were excellent or good for 73% of the patients in the nail group versus 35% in the pin group. The lesions treated were large, with an average size of 447 mm(2), affecting the weightbearing area in the majority of the patients. On magnetic resonance imaging, incomplete bone consolidation was predominant in the pin group. CONCLUSIONS: Arthroscopic fixation with bioabsorbable nails seems to be a suitable method of repair for osteochondritis dissecans of the adult knee and appears to be superior to arthroscopic fixation with bioabsorbable pins. PMID- 17420510 TI - Outcome of patients experiencing progression or relapse after primary treatment with two cycles of chemotherapy and radiotherapy for early-stage favorable Hodgkin's lymphoma. AB - PURPOSE: To evaluate treatment outcome of patients with early-stage favorable Hodgkin's lymphoma (HL) who experience disease relapse after primary treatment with two cycles of chemotherapy followed by radiotherapy (RT). PATIENTS AND METHODS: Of 1,129 patients with early-stage favorable HL enrolled onto the HD7/HD10/HD13 trials of the German Hodgkin Study Group, 42 patients were identified with treatment failure, of whom eight had primary progressive disease, seven had early relapse (< or = 12 months), and 27 had late relapse (> 12 months). We analyzed this group of patients for risk factors, salvage therapy, and treatment outcome. RESULTS: The median age was 41 years (range, 19 to 72 years); 24 patients were male, 15 patients had outfield relapse, 13 patients infield relapse, and nine patients outfield and infield relapse. At relapse, 24 patients were treated with conventional salvage chemotherapy, 14 patients were treated with high-dose chemotherapy followed by autologous stem-cell transplantation, and four patients were treated with RT alone. At 36 months median follow-up, freedom from second treatment failure (FF2F) and overall survival (OS) were 52% and 67%, respectively. According to the prognostic score for relapsed HL (duration of first remission, clinical stage, and anemia at relapse), patients with two or three poor prognostic features had a significantly worse outcome compared with patients with none or one of these factors (P < .05 for FF2F and OS). CONCLUSION: Relapse after primary treatment with two cycles of doxorubicin, bleomycin, vinblastine, and dacarbazine followed by RT is rare. In our analysis, results were influenced by a high treatment-related mortality rate. Additional studies are needed to define the optimal salvage therapy. PMID- 17420511 TI - The role of the number of uninvolved lymph nodes in predicting locoregional recurrence in breast cancer. AB - PURPOSE: To identify groups of early breast cancer patients with substantial risk (10-year risk > 20%) for locoregional failure (LRF) who might benefit from postmastectomy radiotherapy (RT). PATIENTS AND METHODS: Prognostic factors for LRF were evaluated among 6,660 patients (2,588 node-negative patients, 4,072 node positive patients) in International Breast Cancer Study Group Trials I to IX treated with chemotherapy and/or endocrine therapy, and observed for a median of 14 years. In total, 1,251 LRFs were detected. All patients were treated with mastectomy without RT. RESULTS: No group with 10-year LRF risk exceeding 20% was found among patients with node-negative disease. Among patients with node positive breast cancer, increasing numbers of uninvolved nodes were significantly associated with decreased risk of LRF, even after adjustment for other prognostic factors. The highest quartile of uninvolved nodes was compared with the lowest quartile. Among premenopausal patients, LRF risk was decreased by 35% (P = .0010); among postmenopausal patients, LRF risk was decreased by 46% (P < .0001). The 10-year cumulative incidence of LRF was 20% among patients with one to three involved lymph nodes and fewer than 10 uninvolved nodes. Age younger than 40 years and vessel invasion were also associated significantly with increased risk. Among patients with node-positive disease, overall survival was significantly greater in those with higher numbers of uninvolved nodes examined (P < .0001). CONCLUSION: Patients with one to three involved nodes and a low number of uninvolved nodes, vessel invasion, or young age have an increased risk of LRF and may be candidates for a similar treatment as those with at least four lymph node metastases. PMID- 17420512 TI - Patterns of survival in multiple myeloma: a population-based study of patients diagnosed in Sweden from 1973 to 2003. AB - PURPOSE: To define patterns of survival among all multiple myeloma (MM) patients diagnosed in Sweden during a 30-year period. PATIENTS AND METHODS: A total of 14,381 MM patients (7,643 males; 6,738 females) were diagnosed in Sweden from 1973 to 2003 (median age, 69.9 years; range 19 to 101 years). Patients were categorized into six age categories and four calendar periods (1973 to 1979, 1980 to 1986, 1987 to 1993, and 1994 to 2003). We computed relative survival ratios (RSRs) as measures of patient survival. RESULTS: One-year survival improved (P < .001) over time in all age groups and RSRs were 0.73, 0.78, 0.80, and 0.82 for the four calendar periods; however, improvement in 5-year (P < .001) and 10-year (P < .001) RSR was restricted to patients younger than 70 years and younger than 60 years, respectively. For the first time, in analyses restricted to MM patients diagnosed at age younger than 60 years, we found a 29% (P < .001) reduced 10-year mortality in the last calendar period (1994 to 2003) compared with the preceding calendar period (1987 to 1993). Females with MM had a 3% (P = .024) lower excess mortality than males. CONCLUSION: One-year MM survival has increased for all age groups during the last decades; 5-year and 10-year MM survival has increased in younger patients (younger than 60 to 70 years). High-dose melphalan with subsequent autologous stem-cell transplantation, thalidomide, and a continuous improvement in supportive care measures are probably the most important factors contributing to this finding. New effective agents with a more favorable toxicity profile are needed to improve survival further, particularly in the elderly. PMID- 17420513 TI - Rituximab added to first-line mitoxantrone, chlorambucil, and prednisolone chemotherapy followed by interferon maintenance prolongs survival in patients with advanced follicular lymphoma: an East German Study Group Hematology and Oncology Study. AB - PURPOSE: Rituximab has been shown to be active in follicular lymphoma (FL), both as monotherapy and in combination with chemotherapy. We conducted a randomized trial comparing mitoxantrone, chlorambucil, and prednisolone (MCP) chemotherapy plus rituximab with MCP alone. PATIENTS AND METHODS: Previously untreated patients with stage III or IV CD20+ indolent or mantle cell lymphoma were randomly assigned to either eight 28-day cycles of MCP plus rituximab (R-MCP; n = 181) or eight cycles of MCP alone (n = 177). All patients who achieved a complete or partial remission were treated with interferon maintenance until relapse. Herein, we report the results from the primary analysis population of patients with FL, who constituted the majority of patients (56%) recruited to the trial (n = 201; R-MCP, n = 105; MCP, n = 96). RESULTS: Rates of overall and complete response were significantly higher in the R-MCP arm than the MCP arm (overall response, 92% v 75%, respectively; P = .0009; complete response, 50% v 25%, respectively; P = .004). With a median follow-up time of 47 months, median event free survival (EFS) and progression-free survival (PFS) times were significantly prolonged with R-MCP compared with MCP (EFS, not reached v 26 months, respectively; P < .0001; PFS, not reached v 28.8 months, respectively; P < .0001), and overall survival (OS) was significantly improved with R-MCP compared with MCP (4-year OS rate, 87% v 74%, respectively; P = .0096). CONCLUSION: The R MCP regimen significantly improves complete and overall response rates, EFS, PFS, and OS in patients with previously untreated advanced FL, without a clinically significant increase in toxicity. PMID- 17420514 TI - Lethal and nonlethal violence against an intimate female partner: comparing male murderers to nonlethal abusers. AB - Men's lethal and nonlethal violence against an intimate female partner are compared. Various risk factors are examined to compare men's lethal and nonlethal violence against an intimate woman partner. Relative to abusers, men who kill are generally more conventional with respect to childhood backgrounds, education, employment, and criminal careers, are more likely to be possessive and jealous, and are more likely to be separated from their partner at the time of the event. Men who kill are more likely to have used violence against a previous partner, to have sexually assaulted and strangled the victim, and to have used a weapon or instrument. However, they were less likely to have been drunk at the time of the event and/or to have previously used violence against the woman they killed. Overall, the findings do not support the notion of a simple progression from nonlethal to lethal violence and raise some dilemmas for the growing area of risk assessment. PMID- 17420515 TI - Battered pets and domestic violence: animal abuse reported by women experiencing intimate violence and by nonabused women. AB - Women residing at domestic violence shelters (S group) were nearly 11 times more likely to report that their partner had hurt or killed pets than a comparison group of women who said they had not experienced intimate violence (NS group). Reports of threatened harm to pets were more than 4 times higher for the S group. Using the Conflict Tactics Scale, the authors demonstrated that severe physical violence was a significant predictor of pet abuse. The vast majority of shelter women described being emotionally close to their pets and distraught by the abuse family pets experienced. Children were often exposed to pet abuse, and most reported being distressed by these experiences. A substantial minority of S-group women reported that their concern for their pets' welfare prevented them from seeking shelter sooner. This seemed truer for women without children, who may have had stronger pet attachments. This obstacle to seeking safety should be addressed by domestic violence agencies. PMID- 17420516 TI - A comparison of the police response to heterosexual versus same-sex intimate partner violence. AB - It has been argued that the police do not respond to domestic calls involving same-sex couples in the same manner as they respond to calls involving heterosexual couples. A major problem facing researchers examining the police response to cases involving same-sex couples has been the lack of adequately sized samples. In this article, the authors utilize the 2000 National Incident Based Reporting System database, which contains 176,488 intimate partner assaults and intimidation incidents reported to 2,819 police departments in 19 states. The key issue examined is whether similar cases involving same-sex and heterosexual couples result in the same police response. PMID- 17420517 TI - Effects of coordinated services for drug-abusing women who are victims of intimate partner violence. AB - This article summarizes outcomes from a demonstration project on collaboration between substance abuse and domestic violence agencies. Researchers recruited women seeking services for substance abuse or intimate partner violence at 1 of 6 participating agencies. Admitted women were both victims of domestic violence and abusing alcohol or drugs. Following an initial screening, participants were interviewed at program entry (n = 255) and again 4 to 6 months later (n = 128, 50%). Key outcomes were the number of days substances were used in the past 30 days, women's perceptions of harm from battering, and domestic violence self efficacy. Results suggest participants used substances less frequently and experienced themselves as more efficacious following services, but they were also more fearful of the consequences of domestic violence. Repeated-measures MANOVA found that substance abuse days and domestic violence self-efficacy significantly contributed to the multivariate function. Implications for services for women with co-occurring substance abuse and domestic violence victimization are discussed. PMID- 17420518 TI - Barriers to working with sexual assault survivors: a qualitative study of rape crisis center workers. AB - To better understand barriers service providers may face when advocating for survivors, a study using grounded theory and qualitative, semistructured interviews was conducted of rape victim advocates (N= 25) working in rape crisis centers in a large metropolitan area. Broader societal attitudes framed and were reflected in institutional responses to victims and in barriers faced by advocates working with survivors. Organizational barriers noted by advocates related to resources, environmental factors, professionalization, and racism. Staff burnout was a major barrier affecting advocates' ability to help survivors. Finally, the most salient direct service barrier was secondary victimization by criminal justice and medical or mental health systems. PMID- 17420519 TI - Nurse researchers begin as undergraduate students. PMID- 17420520 TI - Prenatal health promotion content and health behaviors. AB - To improve the effectiveness of prenatal care, there is a need to understand the association between health promotion content and health behaviors during pregnancy. The purpose of this study was to determine the association between prenatal health promotion content and health behaviors during pregnancy among low income African American and Mexican American women. Twenty-two recommended health promotion topics and their related health behaviors were measured. Structured interviews on health promotion topics and health behaviors were conducted with 159 pregnant women receiving prenatal care at a low-risk urban clinic. Wide variation was found in the number of health promotion topics discussed. Bivariate and regression analyses examined the association between content topics and behaviors. Healthier behaviors were associated with women's reports of discussing more health promotion topics, using fewer substances prepregnancy, and having a more positive attitude toward pregnancy. Data suggest that increasing prenatal health promotion content during routine visits might improve the effectiveness of prenatal care. PMID- 17420521 TI - Health behaviors of low-income pregnant minority women. AB - Preventive and health promoting behaviors in pregnant minority women can be used to develop approaches to encourage healthy lifestyle and optimal utilization of health services, and to obtain better outcomes of pregnancy. Using the Health Promotion Model as a theoretical framework, this study employed a cross-sectional survey design to investigate factors that related to health behaviors of low income pregnant Mexican American and African American women in selected sites in the southeast portion of the state of Texas. Results showed that Mexican American women had a significantly higher mean on the Powerful Others Locus of Control measure than the African American women. For African American subjects, the strongest effect was exerted by the social support variable, which had a negative effect on the health behavior variable. Findings suggest that more studies are needed to identify specific determinants of health promoting activities during pregnancy in ethnic minority groups. Designing programs that would motivate early involvement of low-income pregnant minority women in health care should be a top priority for professionals in maternity practice. PMID- 17420522 TI - Stages and processes of smoking cessation among adolescents. AB - The purpose of this study was to identify the stages and processes of change related to smoking cessation among vocational technical high school students to provide baseline data for smoking cessation programs. Cross-sectional data were collected in April and May, 2004, from 300 male students at three vocational technical high schools in Korean metropolitan areas. The survey instruments included: decisional balance; self-efficacy; nicotine dependence; stages and processes of change; and knowledge related to smoking. The results indicated that 31.7%, 39.2%, and 29.1% of current smokers were in the precontemplation, contemplation, and preparation stages, respectively, and that 42.6% and 57.4% of former smokers were in the action and maintenance stages, respectively. Decisional balance, self-efficacy, and nicotine dependence were significantly associated with the stages of change. Regarding the processes of change, only behavioral processes were significantly associated with the stages of change, with experiential processes showing no significant relationship. The study results suggest that smoking cessation programs should emphasize behavioral strategies to help students progress through the stages. PMID- 17420523 TI - Measuring elders' symptoms with daily diaries and retrospective reports. AB - The current study investigated the sufficiency of symptom data collected for 7 days from a daily diary and the comparability and validity of data obtained from daily diaries and retrospective reports. Three hundred and five older persons completed a daily symptom diary, a retrospective symptom questionnaire, and a measure of health status. The number of symptoms endorsed each day on the diary declined, F(6, 214) = 13.51, p< .0001. New symptoms were endorsed 15%, and previous symptoms were no longer endorsed 1.9% of the time. More symptoms were endorsed on retrospective reports than on daily diaries, t(304) = 8.48, p < .0001. Symptoms from both methods were significantly correlated with health status (p <.0001) and differences in the correlations were not statistically significant, t(304) = 1.62, p = ns . Because of comparable criterion validity, less burdensome retrospective reports should be used, unless the focus is new symptoms or how symptoms unfold over time. PMID- 17420524 TI - A meta-analysis of interventions for informal stroke caregivers. AB - The purpose of this study is to examine the effectiveness of the interventions for improving mental health of caregivers of people with stroke by synthesizing individual studies. A meta-analysis was performed to summarize findings of intervention studies of caregivers of elderly stroke patients. Additionally, a sensitivity analysis and a publication bias were tested. The overall mean weighted effect size (MWES) for the four studies is 0.277 (Z = 3.432, p = .001) with a 95% CI .118 to .435 (N = 718) indicating that the intervention was effective in improving the mental health of informal stroke caregivers. The MWES for the education program was 0.354 (Z= 2.613, p < .01) and for the support program was .234 (Z = 2.335, p= .02). The MWES for the Europe study was .219 (Z= 2.613, p < .01) and for the United States was .922 (Z = 3.287, p = .001). The results show that overall interventions improved mental health of informal stroke caregivers. The small number of studies included in the meta-analysis limits the generalizability of results while supporting the need for more research in this area. PMID- 17420525 TI - Abused African American women's processes of staying healthy. AB - Fifteen African American women with a history of intimate partner violence (IPV) were interviewed to examine (a) the ways in which poor, urban African American women stay healthy, and particularly how they protected themselves from sexually transmitted diseases and HIV while in abusive relationships; and (b) the roles of intersecting contextual factors such as lifetime experiences of violence, mental health symptoms, and substance use in women's processes of maintaining their health. Data were analyzed using a qualitative descriptive approach. Women were managing numerous, complex problems as they actively worked to maintain their mental and physical health and that of their children. The turning point at which women made substantial changes came when women were "tired" and believed that a new beginning was needed. Racism, poverty, multiple experiences of violence, and health and mental health problems influenced women's health care decisions. Women's health maintenance strategies were often not visible to health care providers and included some behaviors that may place women at greater risk of violence or disease from the point of view of the health care provider. PMID- 17420526 TI - Characterization of protein transacetylase from human placenta as a signaling molecule calreticulin using polyphenolic peracetates as the acetyl group donors. AB - We have earlier shown that a unique membrane-bound enzyme mediates the transfer of acetyl group(s) from polyphenolic peracetates (PA) to functional proteins, which was termed acetoxy drug: protein transacetylase (TAase) because it acted upon several classes of PA. Here, we report the purification of TAase from human placental microsomes to homogeneity with molecular mass of 60 kDa, exhibiting varying degrees of specificity to several classes of PA confirming the structure activity relationship for the microsome-bound TAase. The TAase catalyzed protein acetylation by a model acetoxy drug, 7,8-diacetoxy-4-methyl coumarin (DAMC) was established by the demonstration of immunoreactivity of the acetylated target protein with anti-acetyl lysine antibody. TAase activity was severely inhibited in calcium-aggregated microsomes as well as when Ca2+ was added to purified TAase, suggesting that TAase could be a calcium binding protein. Furthermore, the N-terminal sequence analysis of purified TAase (EPAVYFKEQFLD) using Swiss Prot Database perfectly matched with calreticulin (CRT), a major microsomal calcium binding protein of the endoplasmic reticulum (ER). The identity of TAase with CRT was substantiated by the observation that the purified TAase avidly reacted with commercially available antibody raised against the C-terminus of human CRT (13 residues peptide, DEEDATGQAKDEL). Purified TAase also showed Ca2+ binding and acted as a substrate for phosphorylation catalyzed by protein kinase C (PKC), which are hallmark characteristics of CRT. Further, purified placental CRT as well as the commercially procured pure CRT yielded significant TAase catalytic activity and were also found effective in mediating the acetylation of the target protein NADPH cytochrome P-450 reductase by DAMC as detected by Western blot using anti-acetyl lysine antibody. These observations for the first time convincingly attribute the transacetylase function to CRT. Hence, this transacetylase function of CRT is designated calreticulin transacetylase (CRTAase). We envisage that CRTAase plays an important role in protein modification by way of acetylation independent of Acetyl CoA. PMID- 17420527 TI - Introduction: new frontiers in education and training for the laboratory animal community and public: an overview of select proceedings from the June 2006 Forum of the American College of Laboratory Animal Medicine. PMID- 17420528 TI - Invited international perspective: education and training for the care and use of laboratory animals: an overview of current practices. PMID- 17420529 TI - Training in the laboratory animal science community: strategies to support adult learning. AB - The essence of learning is change; learning is the process by which learners customize new information to make it personally meaningful and relevant. Training is the process of helping students make those changes. Research indicates that adults learn differently than children or adolescents and that adults consistently use the following six learning strategies: prior experiences; conversations; metacognition; reflection; authentic experiences; and images, pictures, or other types of visuals. Each of these learning strategies can be combined with the other strategies and often build upon each other. A recent study on how health care professionals learn indicated that the learning strategy they used most often was reflection, which supports learning before, during, and after training. Numerous examples are provided in this article describing how to integrate each of the six adult learning strategies into laboratory animal science training. While lectures and other types of direct instruction are appropriate, they are inadequate and ineffective unless they are integrated with and support adult learning strategies. Both the US Department of Agriculture regulations and the Public Health Service Policy mandate that research institutions must ensure that all personnel involved in animal care, treatment, or use are qualified to perform their duties. Applying adult learning strategies to training for the laboratory animal science community will enhance learning and improve both the science and the humane care of the animals, which is a goal our community must continuously strive to achieve. PMID- 17420530 TI - Institutional and IACUC responsibilities for animal care and use education and training programs. AB - Training and instruction of personnel are important components of animal care and use programs because they help to ensure the health and welfare of the animals and the integrity of the research or testing results. Training also helps to promote the consideration of alternatives, recognition of animal pain and distress, appropriate use of pain-relieving agents, aseptic technique, pre- and post-procedural care, and personnel health and safety. While individuals who provide the care for or conduct research or testing in laboratory animals should take personal responsibility for ensuring that they have the skills to perform their duties, the institution is ultimately responsible for ensuring their competency. The institution is also responsible for providing the training or instruction that is required by federal legislation, regulations, and policies. The institutional animal care and use committee (IACUC) is responsible for ensuring, as part of their review of research activities, that the personnel are capable of performing the procedures described. The IACUC must also assess the institution's training program as part of their semiannual animal care and use program review and make recommendations regarding training to the institutional official. This article provides a comprehensive overview of the US regulatory mandates for training and personnel qualification. PMID- 17420531 TI - Fundamental training for individuals involved in the care and use of laboratory animals: a review and update of the 1991 NRC Core Training Module. AB - Public trust demands that individuals who do research, testing, or teaching with animals use humane, ethical, and scientifically sound methods. Furthermore, the Animal Welfare Act and the Public Health Service Policy require research institutions to provide basic training and to ensure that anyone who cares for and/or works with laboratory animals has the appropriate training or experience relevant to their job responsibilities. Institutions accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International must also provide training programs and ensure the qualifications of personnel. The primary goal of this training is to provide individuals with basic knowledge and to reinforce attitudes and behaviors that help to ensure humane animal care and use. This article provides an overview of the core training module outline and content from the 1991 report of the Institute for Laboratory Animal Research, Education and Training in the Care and Use of Laboratory Animals: A Guide for Developing Institutional Programs, as well as pertinent updates for introducing personnel to information regarding the care and use of laboratory animals. Both mandatory and suggested training topics are reviewed, including relevant regulations and standards, ethical considerations, humane methods of animal experimentation and maintenance, and other pertinent topics. Although the fundamental training course content and delivery will vary depending on the nature and complexity of an institution's animal care and use program, this basic training provides the foundation for more in-depth training programs and supports humane and ethical animal care and use. PMID- 17420532 TI - Training strategies for animal care technicians and veterinary technical staff. AB - An institutional training program for animal care and veterinary technicians should be planned and implemented to provide these individuals with knowledge and skills for performing their duties within a laboratory animal care and use program. The complexity in the regulatory and scientific features of the animal research environment necessitates a strong training program on diverse topics according to staff duties. Orientation training should include ethics and compliance with relevant laws, policies, and guidelines. Depending on specific staff responsibilities, training may be general or in depth on topics of species specific biology and behavior, animal facility equipment and operations, animal health procedures, animal research policies, occupational health and safety equipment and practices, computer usage, training, and management. Staff training should be an ongoing mission for incorporating new equipment, practices, and procedures in the laboratory animal program; for providing periodic refresher training to maintain a high level of staff qualifications; and for retraining when skills or knowledge are found deficient. Large institutions often have a dedicated training staff to implement the institutional training program. PMID- 17420533 TI - Training strategies for research investigators and technicians. AB - Training programs for research personnel are discussed as a key resource that must be part of an effective animal care and use program. Because of the legal responsibility to ensure that research staff are qualified to use animals, many institutions have justified the necessity for a training coordinator and/or trainers for their animal care and use programs. Effective training programs for research personnel must meet the needs of the client base (research scientists and staff) so that they are relevant, practical, and timely. To meet these objectives, it is useful to involve the scientific staff in the analysis of their learning needs. To meet a performance standard necessary for quality research, a large percentage of the institutional staff must participate in the training program. Often it is the principal investigators who set the tone for their staff members regarding the importance of receiving training. Garnering support from this client base will create a culture that encourages training and engenders a positive attitude about humane animal care and use. One effective approach is to incorporate nonanimal models as alternatives to live animals to teach humane handling techniques and methods, thereby contributing to refinement, reduction, and replacement (the 3Rs). Also discussed are the necessity of timely feedback from clients, documentation of personnel training for regulatory purposes, and the collection of training metrics, which assists in providing justification for the granting of additional fiscal support for the program. Finally, the compliance procedures and opportunities for essential refresher training are discussed and related to high performance standards, humane animal use, and quality research, all of which contribute to the 3Rs. PMID- 17420534 TI - Training strategies for IACUC members and the institutional official. AB - Institutions are required by federal laws and regulations to oversee and evaluate their programs, facilities, and procedures for using animals in research, teaching, and/or testing activities. These responsibilities are specifically charged to an institutional official (IO) and an institutional animal care and use committee (IACUC). Initially, the individuals tasked with these responsibilities seldom have the requisite knowledge or experience to fulfill their charges effectively. Furthermore, simply reading the regulatory requirements does not prepare the novice IO and IACUC members to effectively monitor and guide the program. As a result, many new IOs and IACUC members are managing their responsibilities with insufficient understanding of the laws, regulations, standards, and policies. Specific training strategies for inexperienced IACUC members are needed to help them understand their responsibilities for ensuring animal welfare through an effective, high-quality, and compliant animal care and use program that supports the critical research needed to improve human and animal health. Likewise, most IOs would benefit from training to help them better understand their responsibility for enhancing or maintaining the quality of the institution's animal care and use program. Education and training should begin with an orientation to the laws, regulations, standards, and policies. Continuing training and education are also important to keep abreast of the changes in the interpretation of these laws and regulations as well as the changes in veterinary science. For both the IO and the IACUC, understanding and acceptance of their authority and responsibilities are significant factors in establishing and maintaining a quality animal care and use program. PMID- 17420535 TI - Training strategies for laboratory animal veterinarians: challenges and opportunities. AB - The field of laboratory animal medicine is experiencing a serious shortage of appropriately trained veterinarians for both clinically related and research oriented positions within academia, industry, and government. Recent outreach efforts sponsored by professional organizations have stimulated increased interest in the field. It is an opportune time to critically review and evaluate postgraduate training opportunities in the United States and Canada, including formal training programs, informal training, publicly accessible training resources and educational opportunities, and newly emerging training resources such as Internet-based learning aids. Challenges related to each of these training opportunities exist and include increasing enrollment in formal programs, securing adequate funding support, ensuring appropriate content between formal programs that may have diverse objectives, and accommodating the training needs of veterinarians who enter the field by the experience route. Current training opportunities and resources that exist for veterinarians who enter and are established within the field of laboratory animal science are examined. Strategies for improving formal laboratory animal medicine training programs and for developing alternative programs more suited to practicing clinical veterinarians are discussed. In addition, the resources for high-quality continuing education of experienced laboratory animal veterinarians are reviewed. PMID- 17420536 TI - Evaluating the effectiveness of training strategies: performance goals and testing. AB - The Public Health Service policy, Animal Welfare Act regulations, and the Guide for the Care and Use of Laboratory Animals all require that institutions provide training for personnel engaged in animal research. Most research facilities have developed training programs to meet these requirements but may not have developed ways of assessing the effectiveness of these programs. Omission of this critical activity often leads to training that is ineffective, inefficient, or unnecessary. Evaluating the effectiveness of biomedical research and animal care training should involve a combination of assessments of performance, competence and knowledge, and appropriate tests for each type of knowledge, used at appropriate time intervals. In this article, the hierarchical relationship between performance, competence, and knowledge is described. The discussion of cognitive and psychomotor knowledge includes the important distinction between declarative and procedural knowledge. Measurement of performance is described and can include a variety of indirect and direct measurement techniques. Each measurement option has its own profile of strengths and weaknesses in terms of measurement validity, reliability, and costs of development and delivery. It is important to understand the tradeoffs associated with each measurement option, and to make appropriate choices of measurement strategy based on these tradeoffs arrayed against considerations of frequency, criticality, difficulty of learning, logistics, and budget. The article concludes with an example of how these measurement strategies can be combined into a cost-effective assessment plan for a biomedical research facility. PMID- 17420537 TI - Emerging technologies in education and training: applications for the laboratory animal science community. AB - This article examines several new and exciting communication technologies. Many of the technologies were developed by the entertainment industry; however, other industries are adopting and modifying them for their own needs. These new technologies allow people to collaborate across distance and time and to learn in simulated work contexts. The article explores the potential utility of these technologies for advancing laboratory animal care and use through better education and training. Descriptions include emerging technologies such as augmented reality and multi-user virtual environments, which offer new approaches with different capabilities. Augmented reality interfaces, characterized by the use of handheld computers to infuse the virtual world into the real one, result in deeply immersive simulations. In these simulations, users can access virtual resources and communicate with real and virtual participants. Multi-user virtual environments enable multiple participants to simultaneously access computer-based three-dimensional virtual spaces, called "worlds," and to interact with digital tools. They allow for authentic experiences that promote collaboration, mentoring, and communication. Because individuals may learn or train differently, it is advantageous to combine the capabilities of these technologies and applications with more traditional methods to increase the number of students who are served by using current methods alone. The use of these technologies in animal care and use programs can create detailed training and education environments that allow students to learn the procedures more effectively, teachers to assess their progress more objectively, and researchers to gain insights into animal care. PMID- 17420539 TI - Microparticle separation with an acoustic-gravity field controlled by phase-shift operation. AB - The elution of particles from a coupled acoustic-gravity channel has been controlled by a phase-shift operation. Two ultrasound transducers pasted onto the top and bottom walls of a separation channel were driven at the same frequency, but with different phases. Changing the phases of the ultrasounds has allowed the formation of a node near the channel wall, and particles are retained in the channel. Two types of particles are separated under an appropriate condition. PMID- 17420540 TI - Development of an ultra-micro sample injector for gas chromatography using an ink jet microchip. AB - An ultra-micro sample injector for gas chromatography (GC) was developed. An ink jet microchip, originally used for industrial recorder, was modified at the edge near to an orifice, and fixed into the GC. In order to evaluate the characteristics of this injector, a sample injector and a thermal conductive detector (TCD) were connected directly, while water was used as the test sample. The volume of the droplet, the interval time and the back-pressure to the ink-jet microchip were investigated. Within the range of 1 - 5 nL volume injected sample, the TCD response according to the amount of the sample volume (the volume of one droplet from the ink-jet microchip was about 1 nL) was obtained. A good reproducibility of the peak area was obtained to be about 1.0% of the RSD value. In order to compare the injection method of the ink-jet chip with that using a micro-syringe, the method using the ink-jet chip could introduce 1/1000 of the amount of the sample and gave reproducible results. PMID- 17420541 TI - Development of an NMR interface microchip "MICCS" for direct detection of reaction products and intermediates of micro-syntheses using a "MICCS-NMR". AB - The development of an NMR interface microchip and its applications to the real time monitoring of chemical reactions are described. The microchip device was named "MICCS" (MIcro Channeled Cell for Synthesis monitoring), and the method using it was named "MICCS-NMR". MICCS was inserted into a 5 mm Phi NMR sample tube. Thus standard solution NMR probes without any modifications can be used in MICCS-NMR measurements. A gap between MICCS and the sample tube was filled with a deuterated solvent for an NMR lock. The reaction temperature and reaction time in MICCS can be easily changed by adjusting the temperature of the NMR probe and changing the flow rates, respectively. The effectiveness of the MICCS-NMR was verified in the real-time monitoring of the Wittig reaction. Preliminary data on the direct detection of intermediates of the Grignard reaction is also reported. Besides real-time monitoring of chemical reactions, MICCS-NMR would be useful as a qualitative detection method for microchip-based synthesis. PMID- 17420542 TI - Rapid DNA hybridization analysis using a PDMS microfluidic sensor and a molecular beacon. AB - A rapid DNA analysis has been developed based on a fluorescence intensity change of a molecular beacon in a PDMS microfluidic channel. Recently, we reported a new analytical method of DNA hybridization involving a PDMS microfluidic sensor using fluorescence energy transfer (FRET). However, there are some limitations in its application to real DNA samples because the target DNA must be labelled with a suitable fluorescent dye. To resolve this problem, we have developed a new DNA microfluidic sensor using a molecular beacon. By monitoring the change in the restored fluorescence intensity along the channel length, it is possible to rapidly detect any hybridization of the molecular beacon to the target DNA. In this case, the target DNA does not need to be labelled. Our experimental results demonstrate that this microfluidic sensor using a molecular beacon is a promising diagnostic tool for rapid DNA hybridization analysis. PMID- 17420543 TI - Chemiluminescence detection of five-membered heteroaromatic compounds using electrogenerated tris(2,2'-bipyridine)-ruthenium(III). AB - Heteroaromatic compounds, such as 2,5-dimethylthiophene, 2,5-dimethylpyrrole, and 2,5-dimethylfuran, were found to act as reducing agents for the tris(2,2' bipyridine)ruthenium(III), Ru(bpy)3(3+), chemiluminescent reaction. In order to characterize the chemiluminescent reaction of Ru(bpy)3(3+) with heteroaromatic compounds, we have investigated various mono-, di-, and tri-heteroaromatic compounds. The pi-electron density and stability of aromatic rings influence the chemiluminescent efficiency of the reaction. Above all, 2,5-dimethylthiophene produced strong chemiluminescence under acidic conditions. In addition, we confirmed that the rate of the chemiluminescent reaction of Ru(bpy)3(3+) with 2,5 dimethylthiophene is very fast. PMID- 17420544 TI - Development of novel reagent for Hantzsch reaction for the determination of formaldehyde by spectrophotometry and fluorometry. AB - A novel reagent, acetoacetanilide (AAA), was introduced to the determination of formaldehyde based on Hantzsch reaction. A simple and highly sensitive fluorometric method was achieved by using AAA. The main advantages in the use of this reagent are: the reaction is carried out at room temperature without any heating system, the cyclization product based on Hantzsch reaction is soluble in water, and the product can be detected by spectrophotometry and fluorometry. The maximum absorption wavelength of the product occurs at 368 nm, and the maximum excitation and emission wavelengths are found at 370 and 470 nm, respectively. Several important experimental variables of the procedures were examined; particularly, the reaction temperature, reaction time, concentrations of reagents, and pH of the reagent solution were optimized for improving the detecting sensitivity. The calibration graph was linear in the range of 1 x 10( 7) - 1 x 10(-6) M or much higher concentrations. The limit of detection (LOD), based on three times of the standard deviation of the reagent blank, was 2.0 x 10(-8) M. The proposed method was applied to the determination of formaldehyde in environmental water samples. Many foreign species commonly existing in water samples did not interfere with the determination of formaldehyde in the proposed method. PMID- 17420545 TI - Interactions of lysozyme with 6-amino-4-aryl-3-methyl-1-phenyl-1H-pyrazolo[3,4 b]pyridine-5-carbonitriles: a fluorescence quenching study. AB - The interactions between 6-amino-4-aryl-3-methyl-1-phenyl-1H-pyrazolo[3,4 b]pyridine-5-carbonitrile and lysozyme (LYSO) were investigated by using tryptophane fluorescence quenching and 6-amino-4-(2-hydroxyphenyl)-3-methyl-1 phenyl-1H-pyrazolo[3,4-b]pyridine-5-carbonitrile (1) was studied in detail because of its high water solubility. At different temperatures, the quenching constants K(SV), the binding constants K and the binding sites n of LYSO with 1 were determined and the thermodynamic parameters were calculated. The distance r between tryptophane residues and 1 was obtained according to the Forster mechanism of non-radiation energy transfer. Furthermore, synchronous fluorescence spectroscopy data indicated that the association between 1 and LYSO changed LYSO's conformation and that the hydrophobic interaction played a major role in 1 LYSO association. It was proved that the fluorescence quenching of LYSO by 1 was related to the formation of a 1-LYSO complex and to a non-radiation energy transfer. PMID- 17420546 TI - Multicommuted fluorometric multiparameter sensor for simultaneous determination of naproxen and salicylic acid in biological fluids. AB - A combined approach based on solid-phase optosensing and multicommutation principles has been applied to develop a method for the simultaneous analysis of two pharmaceuticals (naproxen and salicylic acid) in biological fluids. The multicommuted flow-through optosensor was based on direct native fluorescence measurements of both analgesics using a non-polar sorbent (C18 silica gel) as a solid sensing zone. The flow system was controlled by Java-written home-made software and designed using three-way solenoid valves for an independent automated manipulation of sample and carrier solutions. Using an optimized sampling time, the method was calibrated in the range of 1 - 25 and 5 - 200 ng mL(-1). The obtained detection limits were 0.3 and 1.3 ng mL(-1) for naproxen and salicylic acid, respectively, with RSD (%) values of better than 2% for both analytes. The proposed methodology was successfully applied to urine, serum and pharmaceutical preparations. Recovery percentages ranging from 96.1 to 104% were obtained for both analytes. PMID- 17420547 TI - Interaction of tetrandrine with human serum albumin: a fluorescence quenching study. AB - The interaction of tetrandrine with human serum albumin (HSA) was studied by measuring fluorescence quenching spectra, synchronous fluorescence spectra and ultra-violet spectra. The fluorescence quenching spectra of HSA in the presence of tetrandrine showed that tetrandrine quenched the fluorescence of HSA. The quenching constants of tetrandrine on HSA were determined using the Stern-Volmer equation. Static quenching and non-radiation energy transfer were the two main reasons leading to the fluorescence quenching of HSA by tetrandrine. According to the Forster theory of non-radiation energy transfer, the binding distances (r) and the binding constants (K(A)) were obtained. The thermodynamic parameters obtained in this study revealed that the interaction between tetrandrine and HSA was mainly driven by a hydrophobic force. The conformational changes of HSA were investigated by synchronous spectrum studies. PMID- 17420548 TI - Multivariate classification of cigarettes according to their elemental content determined by inductively coupled plasma optical emission spectrometry. AB - In this paper we describe our study on the characterization of cigarette samples according to their mineral content. Acid digestion assisted by microwaves was employed, and inductively coupled plasma optical emission spectrometry was the analytical technique used for the determination of Al, Ba, Ca, Cu, Fe, K, Mg, Mn, Na, P and Sr in conventional, light, and flavorized cigarettes. Multivariate techniques, such as hierarchical clusters analysis (HCA) and principal-component analysis (PCA), were applied to discriminate among different types of cigarettes. Cluster analysis and principal-component analysis showed differences in cigarettes according to the type and mineral composition. The cigarette samples were divided within the 3 groups according to their mineral composition. Ca, Sr, Cu, K and Na were the most important elements for cigarette classification, and only these 5 variables were sufficient for the classification and discrimination of the evaluated types of cigarettes. PMID- 17420549 TI - Integrated rate equation considering product inhibition and its application to kinetic assay of serum ethanol. AB - Kinetic assay of serum ethanol was investigated by predicting maximal product absorbance at 340 nm (A(mk)) through fitting to the yeast alcohol dehydrogenase reaction curve with the integrated rate equation, taking into account product inhibition in the presence of semicarbazide. Predicted A(mk) linearly responded to the preset constant of steady-state concentration of acetaldehyde (C(ald)). An exponential correlation function was established between desired C(ald) and putative A(mk) for authentic ethanol. For unknown samples, iterative fitting to reaction curve till preset constant C(ald) and resultant A(mk) satisfied this exponential correlation function yielded A(mk) with variation coefficient <4.3%. Variations in enzyme activity, data range and kinetic parameters showed negligible effects. The recovery was consistent to 100% with resistance to methanol and isopropanol. The upper limit of linear response for A(mk) was about 40 times of the lower limit. These results indicated that this kinetic method was reliable for serum ethanol assays with obvious advantages. PMID- 17420550 TI - Simultaneous quantitative resolution of atorvastatin calcium and fenofibrate in pharmaceutical preparation by using derivative ratio spectrophotometry and chemometric calibrations. AB - In the present work, five different spectrophotometric techniques for simultaneous determination of formulations containing atorvastatin calcium (ATOR) and fenofibrate (FENO) in various combinations are described. In ratio spectra derivative spectrophotometry, analytical signals were measured at wavelengths corresponding to either maximums or minimums for both drugs in first derivative spectra of ratio spectra obtained by using either spectrum as divisor. For the remaining four methods using chemometric techniques, namely, classical least squares (CLS), inverse least squares (ILS), principal component regression (PCR) and partial least squares (PLS), the calibrations were constructed by using the absorption data matrix corresponding to the concentration data matrix, with measurements in the range of 231 - 310 nm (Deltalambda = 1 nm) in their zero order spectra. The linearity range was found to be 4 - 22 and 2 - 20 microg/ml for ATOR and FENO, respectively. The validity of the proposed methods was successfully assessed for analyses of both drugs in laboratory-prepared mixtures and in commercial tablet formulations. PMID- 17420551 TI - Simultaneous fluorometric resolution of pyridoxal, pyridoxamine and 4-pyridoxic acid using chemometric methods. AB - An alternatively minimizing covariant matrix error (AMCME) algorithm, newly proposed by the present authors, was applied to the simultaneous fluorometric determination of pyridoxal, pyridoxamine and 4-pyridoxic acid without loss of sensitivity. The experimental results illustrate that the profiles of spectra and concentration can be accurately resolved using the AMCME algorithm with a high sensitivity and stable repeatability. That is to say, the closely overlapping problem of the spectra could be resolved owing to the characteristic features of the AMCME algorithm. PMID- 17420552 TI - Photothermal techniques applied to the determination of the water vapor diffusion coefficient and thermal diffusivity of edible films. AB - Water vapor diffusion coefficient (WVDC) and thermal diffusivity (alpha) were determined in gelatin-starch films through photothermal techniques. The effect of different variables in the elaboration of these films, such as starch and glycerol concentrations and pH, were evaluated through the response surface methodology. The results indicated that an increase in the glycerol concentration and pH favored the WVDC of the films. On the other hand, alpha was influenced principally by the starch content and pH of the film-forming solution. The minimum alpha value was 4.5 x 10(-4) cm2/s, which is compared with alpha values reported for commercial synthetic polymers. PMID- 17420553 TI - A wireless pH sensor using magnetoelasticity for measurement of body fluid acidity. AB - The determination of body fluid acidity using a wireless magnetoelastic pH sensitive sensor is described. The sensor was fabricated by casting a layer of pH sensitive polymer on a magnetoelastic ribbon. In response to an externally applied time-varying magnetic field, the magnetoelastic sensor mechanically vibrates at a characteristic frequency that is inversely dependent upon the mass of the pH polymer film, which varies as the film swells and shrinks in response to pH. As the magnetoelastic sensor is magnetostrictive, the mechanical vibrations of the sensor launch magnetic flux that can be detected remotely using a pickup coil. The sensor can be used for direct measurements of body fluid acidity without a pretreatment of the sample by using a filtration membrane. A reversible and linear response was obtained between pH 5.0 and 8.0 with a measurement resolution of pH 0.1 and a slope of 0.2 kHz pH(-1). Since there are no physical connections between the sensor and the instrument, the sensor can be applied to in vivo and in situ monitoring of the physiological pH and its fluctuations. PMID- 17420554 TI - A highly selective and sensitive thorium(IV) PVC membrane electrode based on a dithio-tetraaza macrocyclic compound. AB - A poly(vinyl chloride)-based membrane composed of dithio-tetraaza macrocyclic compound as a neutral carrier with sodium tetraphenylborate (NaTPB) as an anion excluder and nitrobenzene (NB) as plasticizer was prepared and investigated as a Th(IV)-selective electrode. The electrode exhibits a Nernstian slope of 14.2 +/- 0.3 mV per decade over a wide concentration range (1.0 x 10(-6) to 1.0 x 10(-1) M) with a detection limit of 8.0 x 10(-7) M between pH 3.5 and 9.5. The response time of the sensor is about 10 s and it can be used over a period of 5 months without any divergence in potential. The proposed membrane sensor revealed a good selectivity for Th(IV) over a wide variety of other metal ions and proved to be a better electrode in many respects than those reported in the literature. It was successfully applied as an electrode indicator as well as in the direct determination of thorium ions in standard and real samples. PMID- 17420555 TI - Enhanced electrochemical detection of ketorolac tromethamine at polypyrrole modified glassy carbon electrode. AB - A glassy carbon electrode modified with a coating of polypyrrole (Ppy) exhibited an attractive performance for the detection and determination of a non-steroidal and non-narcotic analgesic compound, ketorolac tromethamine (KT). Cyclic voltammetry, differential pulse and square wave voltammetry were used in a combined way to identify the electrochemical characteristics and to optimize the conditions for detection. For calibrating and estimating KT, square-wave voltammetry was mainly used. The drug shows a well-defined peak at -1.40 V vs. Ag/AgCl in the acetate buffer (pH 5.5). The existence of Ppy on the surface of the electrode gives higher electrochemical active sites at the electrode for the detection of KT and preconcentrate KT by adsorption. The square-wave stripping voltammetric response depends on the excitation signal and the accumulation time. The calibration curve is linear in the range 1 x 10(-11) to 1 x 10(-7) M with a detection limit of 1.0 x 10(-12) M. Applicability to serum samples was also demonstrated. A detection limit of 1.0 ng ml for serum was observed. Square-wave voltammetry shows superior performance over UV spectroscopy and other techniques. PMID- 17420556 TI - Electrochemical behavior and determination of clozapine on a glassy carbon electrode modified by electrochemical oxidation. AB - The adsorptive and electrochemical behaviors of clozapine (CLZ) were investigated on a glassy carbon electrode that was electrochemically treated by anodic oxidation at +1.8 V, following potential cycling in the potential range from -0.8 to 1.0 V vs. Ag/AgCl reference electrode. Based on the obtained electrochemical results, an electrochemical-chemical (EC) mechanism was proposed to explain the electrochemical oxidation of CLZ. The resulting electrochemically pretreated glassy carbon electrode (EPGCE) showed good activity to improve the electrochemical response of the drug. CLZ was accumulated in a phosphate buffer (pH 6) at a certain time, and then determined by differential pulse voltammetry. The anodic and cathodic peak currents showed a linear function in the concentration ranges of 0.1 - 1, 1 - 10 and 10 - 100 microM with various accumulation times. The proposed method was successfully used for the determination of CLZ in pharmaceutical preparations. The preconcentration medium exchange approach was utilized for the selective determination of the drug in spiked urine samples with satisfactory results. The recovery levels of the method reached 96% (RSD, 1.8%) and 90% (RSD, 2.8%) for urine and plasma samples, respectively. PMID- 17420557 TI - Determination of catecholamines and indoleamines in human urine based on intramolecular excimer-forming derivatization and fluorescence detection. AB - A liquid chromatographic (LC) determination of catecholamines and indoleamines is described. This is based on intramolecular excimer-forming fluorescence derivatization with 4-(1-pyrene)butanoyl chloride, followed by reversed-phase LC. The analytes, containing an amino moiety and phenolic hydroxyl moieties in a molecule, were converted to the corresponding polypyrene-labeled derivatives by one-step derivatization. They afforded intramolecular excimer fluorescence, which can clearly be discriminated from the normal fluorescence emitted from reagent blanks. The detection limits (S/N = 3) for catecholamines and indoleamines were femto-mole levels per 20-microL injection. Furthermore, this method was applied to a urine assay. PMID- 17420558 TI - Application of retention factors in affinity electrokinetic chromatography and capillary electrophoresis. AB - Affinity capillary electrophoresis has become an established approach for performing interaction studies. In affinity electrokinetic chromatography the retention factor, as in liquid chromatography, is useful for describing the migration behavior of the analytes, and is instrumental for assessing the affinity of an analyte for the pseudo-stationary phase. Erroneous use of the retention factor concept in affinity capillary electrophoretic studies has appeared in a number of recent papers. The errors and their origin are pointed out, and the correct use of retention factors in affinity electrokinetic chromatography and capillary electrophoresis is summarized. PMID- 17420559 TI - Determination of copper and iron using [S,S']-ethylenediaminedisuccinic acid as a chelating agent in wood pulp by capillary electrophoresis. AB - A capillary electrophoresis (CE) method is described for the simultaneous determination of copper and iron after complexation with a readily biodegradable chelating agent, [S,S']-ethylenediaminedisuccinic acid (EDDS), in wood pulp. CE separation was performed in a fused-silica capillary (50 microm i.d.; total length, 65 cm) with an electrolyte containing 25 mM borate buffer and 0.5 mM CTAB at pH 7.0 and an applied voltage of -25 kV. The samples were introduced by applying a 50 mbar pressure for 2 s, and detection of the complexes was monitored at 245 nm. The methodology performance of the methods was evaluated in terms of the linearity, limit of detection (LOD), limit of quantitation (LOQ) and reproducibility. The applicability of the method was demonstrated for the analysis of copper and iron in wood pulp. PMID- 17420560 TI - Evaluation of a passive air sampler for measuring indoor formaldehyde. AB - A passive air sampler, using 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole, was evaluated for the determination of formaldehyde in indoor environments. Chromatography paper cleaned using a 3% hydrogen peroxide solution was experimentally determined as being the optimum absorption filter for the collection of formaldehyde (0.05 microg cm(-2) formaldehyde). From a linear regression analysis between the mass of formaldehyde time-collected on a passive air sampler and the formaldehyde concentration measured by an active sampler, the sampling rate of the passive air sampler was 1.52 L h(-1). The sampling rate, determined for the passive air sampler in relation to the temperature (19 - 28 degrees C) and the relative humidity (30 - 90%), were 1.56 +/- 0.04 and 1.58 +/- 0.07 L h(-1), respectively. The relationship between the sampling rate and the air velocity was a linear-regression within the observed range. In the case of exposed samplers, the stability of the collected formaldehyde decreased with increasing storage time (decrease of ca. 25% after 22 days); but with the unexposed samplers the stability of the blank remained relatively unchanged for 7 days (decrease of ca. 37% after 22 days). The detection limits for the passive air sampler with an exposure time of 1 day and 7 days were 10.4 and 1.48 microg m(-3), respectively. PMID- 17420561 TI - Azalides from azithromycin to new azalide derivatives. AB - Azalides are semi-synthetic macrolides, in which a nitrogen atom is introduced into a macrolactone ring via a Beckmann rearrangement. Starting from erythromycin, oximes, depending on the reaction conditions lactams, or bicyclic imino-ethers were formed, which were further reduced to aminolactones. The cyclic amine 9a- became the precursor for novel, significantly more active derivatives, especially for 9-dihydro-9-deoxo-9a-methyl-9a-aza-9a-homoerythromycin A with the generic name azithromycin. It showed a broad spectrum of antibacterial activity covering all significant bacteria causing respiratory tract infections. The greatest advantages of azithromycin are its unusual pharmacokinetics (high tissue distribution), metabolic stability and high tolerability. These properties have led in recent years to the widespread use of the azalide scaffold for the synthesis of new compounds with advantageous pharmacokinetics. The azalide scaffold possesses an amino and several hydroxyl groups, which could be substituted or transformed to obtain new compounds. Different derivatives were obtained by substitution on the nitrogen but a large variety of derivatives, such as ethers, esters and carbamates, were made by reactions with various hydroxyl groups. Substitutions on both nitrogen and hydroxyl or two hydroxyl groups yielded new, bridged compounds. The 4''-hydroxy group was oxidized to 4-oxo-, which was transformed via the oxime to 4-amino, or via epoxide to 4''-methylamino compounds. Cleavage of the cladinose sugar and further transformations gave 3 acyl or 3-oxo compounds, which were less active than 14-membered acylides or ketolides. Beckmann rearrangement of some 16-membered macrolide oximes yielded only 17-membered lactams, which were less active than starting macrolides, and could not be reduced to amines. Intramolecular rearrangement of azalide imino ethers yielded 13-membered azalides. Some new 11a-azalides were obtained after oxidative cleavage of some 16-membered macrolides and additional cyclisation. PMID- 17420562 TI - Discovery of 4-Pyridone derivatives as specific inhibitors of enoyl-acyl carrier protein reductase (FabI) with antibacterial activity against Staphylococcus aureus. AB - 4-Pyridone derivatives were identified as potent inhibitors of FabI, the enoyl acyl carrier protein reductase in Escherichia coli and Staphylococcus aureus. 1 Substituted derivatives of a hit compound exhibited potent antibacterial activities against S. aureus. Target specificity of 4-pyridone derivatives was confirmed by the strong inhibition of lipid synthesis in macromolecular biosynthesis assay and also by the reduced antimicrobial activity against triclosan-resistant S. aureus isolates possessing a point mutation (Ala95Val) in FabI. Two 4-pyridone compounds exhibited strong antibacterial activities against 30 clinical isolates of methicillin-resistant S. aureus (MRSA) with MIC(90) of 0.5 and 2 mug/ml, respectively. Moreover, they retained activity against S. aureus with a mutation affecting FabI residue 204, which was recently found to be associated with triclosan resistance in clinical isolates of S. aureus. In conclusion, we have identified a novel chemical series, 4-pyridone derivatives, as specific inhibitors of FabI with potent antibacterial activity against S. aureus. PMID- 17420563 TI - The mode of action of 2-(thiazol-2-ylthio)-1beta-methylcarbapenems against Pseudomonas aeruginosa: the impact of outer membrane permeability and the contribution of MexAB-OprM efflux system. AB - The mode of action of a series of 2-(4-dihydropyrrolylthiazol-2-ylthio) and 2-(4 tetrahydropyridinylthiazol-2-ylthio)-1beta-methylcarbapenem analogues against Pseudomonas aeruginosa was investigated with regard to contributions of the affinity for penicillin binding proteins (PBPs), the outer membrane permeability, and the effect of the MexAB-OprM efflux system. In this series of carbapenems, the introduction of a substituent in C-2 side chain with a change in physicochemical properties affected the antipseudomonal activity depending on the molecular weight. However, these structural modifications did not affect the affinity for pseudomonal PBPs significantly. It was confirmed that the affinity for PBPs was not an important determinant of the antipseudomonal activity of this series of carbapenems. OprD porin-deficiency did not affect antipseudomonal activity either. On the other hand, the MIC of these carbapenems against P. aeruginosa significantly decreased in the presence of outer membrane permeabilizer. This result strongly suggests that the cause of the relatively low antipseudomonal activity of these carbapanems is their low permeability through the outer membrane of P. aeruginosa. And also, in the presence of outer membrane permeabilizer, the MICs against MexAB-OprM deficient mutants remarkably decreased and were very close to the value of the IC(50) for pseudomonal PBPs. From this result, it was clear that the effect of the MexAB-OprM efflux system was also an important determinant of antipseudomonal activity of these carbapenems. In conclusion, the major determinants of the antipseudomonal activity of the 2 (thiazol-2-ylthio)-1beta-methylcarbapenems are the outer membrane permeability and the effect of the MexAB-OprM efflux system, not the affinity for pseudomonal PBPs. PMID- 17420564 TI - Pleofungins, novel inositol phosphorylceramide synthase inhibitors, from Phoma sp. SANK 13899. I. Taxonomy, fermentation, isolation, and biological activities. AB - In the course of a screening for inositol phosphorylceramide (IPC) synthase inhibitors, the novel inhibitors pleofungins A, B, C, and D were found in a mycelial extract of a fungus, Phoma sp. SANK13899. Purification was performed by 50% methanol and ethyl acetate extraction, reversed phase open-column chromatography, and HPLC separations. Pleofungin A inhibited the IPC synthase of Saccharomyces cerevisiae and Aspergillus fumigatus at IC(50) values of 16 and 1.0 ng/ml, respectively. The inhibitor also suppressed the growth of Candida albicans, Cryptococcus neoformans, and A. fumigatus at MIC values of 2.0, 0.3, and 0.5 mug/ml, respectively. These biological properties indicate that pleofungins belong to a novel class of IPC synthase inhibitors efficacious against A. fumigatus. PMID- 17420565 TI - Pleofungins, novel inositol phosphorylceramide synthase inhibitors, from Phoma sp. SANK 13899. II. Structural elucidation. AB - Pleofungins (formerly called F-15078) A, B, C and D, novel depsipeptide antifungal antibiotics, were found in a mycelium extract of the producing fungus, Phoma sp. SANK 13899. The structures of pleofungins A, B, C and D were elucidated mainly by various NMR studies. The absolute configurations of the amino acids and N-methyl amino acids of pleofungin A constituents in the hydrolysate were determined by the application of advanced Marfey's method in combination with gas chromatography/mass spectrometry analysis of their silylation products with N methyl-N-(tert-butylsilyl)trifluoroacetamide. Two alpha-hydroxy acid constituents, alpha-hydroxyisocaproic acid and alpha-hydroxyisovaleric acid, were isolated from the hydrolysate and their stereochemistries were determined by their specific rotations. PMID- 17420566 TI - Carbonarones A and B, new bioactive gamma-Pyrone and alpha-Pyridone derivatives from the marine-derived fungus Aspergillus carbonarius. AB - Two new secondary metabolites, carbonarones A (1) and B (2), were obtained from the culture of the marine-derived fungus Aspergillus carbonarius isolated from the marine sediment collected at Weizhou island of China. Based on ESIMS, 1D and 2D NMR data, and the X-ray crystallographic analysis, their structures were elucidated as 6-benzyl-4-oxo-4H-pyran-3-carboxamide and 6-benzyl-4-hydroxy-2-oxo 1, 2-dihydropyridine-3-carbaldehyde, respectively. 1 and 2 showed moderate cytotoxicity against K562 cells with IC(50) values of 56.0 and 27.8 mug/ml, respectively. PMID- 17420567 TI - A new nucleosidyl-peptide antibiotic, sansanmycin. AB - A new nucleosidyl-peptide antibiotic, sansanmycin, was isolated from an unidentified Streptomyces sp SS. The structure of sansanmycin was elucidated by analyses of its alkaline hydrolysate and spectroscopic analyses. Sansanmycin exhibits antibacterial activity against Mycobacterium tuberculosis H(37)Ra and Pseudomonas aeruginosa with MIC values of 10 and 12.5 mug/ml, respectively. PMID- 17420568 TI - Highly oxidized humulane sesquiterpenes from the basidiomycete Lactarius mitissimus. AB - Two new highly oxidized humulane sesquiterpenes, mitissimols D (1) and E (2) were isolated from the fruiting bodies of Lactarius mitissimus. Their structures were elucidated using extensive spectroscopic techniques including 1D and 2D NMR spectra. PMID- 17420569 TI - Evidence for new beta1-3 galactosyltransferase activity involved in biosynthesis of unusual N-glycan harboring T-antigen in Apis mellifera. AB - In a previous study (Y. Kimura et al., Biosci. Biotechnol. Biochem., 70, 2583 2587, 2006), we found that new complex type N-glycans harboring Thomsen Friedenreich antigen (Galbeta1-3GalNAc) unit occur on royal jelly glycoproteins, suggesting the involvement of a new beta1-3galactosyltransferase in the synthesis of the unusual complex type N-glycans. So far, such beta1-3galactosyltransferase activity, which can transfer galactosyl residues with the beta1-3 linkage to beta1-4 GalNAc residues in N-glycan, has not been found among any eucaryotic cells. But using GalNAc(2)GlcNAc(2)Man(3)GlcNAc(2)-PA as acceptor N-glycan, we detected the beta1-3 galactosyltransferase activity in membrane fraction prepared from honeybee cephalic portions. This result indicates that honeybee expresses a unique beta1-3 galactosyltransferase involved in biosynthesis of the unusual N glycan containing a tumor related antigen in the hypopharyngeal gland. PMID- 17420570 TI - Characterization of the rice circadian clock-associated pseudo-response regulators in Arabidopsis thaliana. AB - Members of the small family of Arabidopsis PSEUDO-RESPONSE REGULATORS (PRR1/TOC1, PRR3, PRR5, PRR7, and PRR9) play roles close to the circadian clock in Arabidopsis thaliana. We have reported that the rice (Oryza sativa) genome also encodes a set of PRR counterparts (designated OsPRR1, OsPRR37, OsPRR59, OsPRR73, and OsPRR95 respectively). To gain new insight into the molecular functions of OsPRRs, we carried out genetic complementation analyses by introducing two representative rice genes, OsPRR1 and OsPRR37, into the corresponding Arabidopsis loss-of-function mutants (toc1 and prr7 respectively). The results showed that these OsPRR and AtPRR genes are genetically interchangeable at least in part, suggesting the conserved clock-associated function of these OsPRRs. PMID- 17420571 TI - Effect of synthetic hydroxy isothiocyanates on a bacterial virus and DNA. AB - The Effect of hydroxy isothiocyanates on a bacterial virus and M13 DNA was examined. Hydroxy-substituted phenyl and phenyl alkyl isothiocyanates, especially 2-(3,4-dihydroxyphenyl)ethyl isothiocyanate(IT-Dop) synthesized from dopamine, showed antiviral activity on psiK. In transfection experiments with M13 mp DNA species, IT-Dop inhibited the single-stranded (SS) molecule more effectively than the double stranded replicative form (RF) DNA. These effects were dependent on reaction time, and on IT-Dop concentration. An additional experiment indicated that treatment with IT-Dop suppressed annealing (reassociation) of denatured DNA. These results indicate that IT-Dop reacts mildly with virus and SS DNA. PMID- 17420572 TI - 5-bromodeoxyuridine increases transient expression of ectopic genes in human cells. AB - Ectopic genes transferred to cells are temporally expressed, although this phenomenon has not yet been well characterized. We found that 5-bromodeoxyuridine dramatically increased transient expression of ectopic genes in human cells. This effect was elicited by adding 5-bromodeoxyuridine prior to or after transfection. No promoter specificity was observed. Real time PCR analysis showed an approximately 2-fold increase in mRNA levels. Since 5-bromodeoxyuridine decondenses heterochromatin and changes the nuclear envelope, these changes might affect transcriptional and post-transcriptional events in the gene expression of plasmids. PMID- 17420573 TI - Human smooth muscle alpha-actin promoter drives Cre recombinase expression in the cranial suture in addition to smooth muscle cell. AB - Tissue-specific gene deletion by the Cre-loxp system is a powerful tool to investigate the roles of specific genes. To determine the specificity and efficiency of the Cre-mediated recombination under the control of the human smooth muscle alpha-actin promoter, we mated SMalphaA-Cre mice and R26R reporter mice. Cre-mediated recombination was observed in visceral and vascular smooth muscle cells. Partial recombination was also found in heart and musculoskeletal connective tissues. Highly efficient recombination was found in cranial sutures. Hence, we propose that SMalphaA-Cre mice are good tool for conditionally deleting gene function in the cranial suture in addition to smooth muscle cells. PMID- 17420574 TI - Identification of in vivo substrates of the chaperonin GroEL from Bacillus subtilis. AB - We investigated GroEL substrates from Bacillus subtilis 168 using the single-ring mutant of B. subtilis GroEL. We identified 28 candidates for GroEL substrates, of which Spo0B, Ald, Eno, SpoIIP, and FbaA were involved in spore formation, and Rnc, Tuf, Eno, Tsf, and FbaA were essential for B. subtilis growth. As observed at the protein level, the amount of SpoIIP interaction with GroEL increased at 3 h after initiation of sporulation. PMID- 17420575 TI - Characterization of dimers of hydroquinone glucosides produced by peroxidase catalyzed polymerization. AB - 4'-Hydroxyphenyl alpha-glucoside and 4'-hydroxyphenyl beta-glucoside were polymerized with horseradish peroxidase. The isolated dimers were found to have linkages at C3' of the hydroxyphenyl moieties and proved to be fluorescent. Low accumulation of oligomers was attributed to increasing electrochemical reactivity with polymerization degrees, which were expected from the levels of highest occupied molecular orbital. PMID- 17420576 TI - Purification, characterization, and molecular cloning of a thermostable superoxide dismutase from Thermoascus aurantiacus. AB - A thermostable superoxide dismutase [(SOD) EC 1.15.1.1] from a Thermoascus aurantiacus var. levisporus was purified to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) homogeneity by a series of column chromatographies. The molecular mass of a single band of the enzyme was estimated to be 16.8 kDa by SDS-PAGE. The molecular mass was estimated to be 33.2 kDa by gel filtration on Sephacryl S-100, indicating that the enzyme was composed of two identical subunits of 16.8 kDa each. N-terminal amino acid sequencing (seven residues) yielded VKAVAVL. Using RACE-PCR, a Cu, Zn-SOD gene was cloned from T. aurantiacus var. levisporus. The sequence was 705 bp and contained a 468 bp ORF encoding a Cu, Zn-SOD of 155 amino acid residues. PMID- 17420577 TI - Synthesis of lipid derivatives of pyrrole polyamide and their biological activity. AB - Novel fatty acyl and phospholipid derivatives of pyrrole polyamide were synthesized. Their cytotoxicity against a cancer cell line of MT-4 cells and those infected by human immunodeficiency virus (HIV) was examined. Although no anti-HIV activity was found, their cytotoxicitty against the cancer cells was significantly enhanced by introducing a lipophilic group into the pyrrole polyamide. PMID- 17420578 TI - Three antinematodal diterpenes from Euphorbia kansui. AB - Three compounds, 20-O-acetyl-[3-O-(2'E,4'Z)-decadienoyl]-ingenol (1), 20-O-acetyl [5-O-(2'E,4'Z)-decadienoyl]-ingenol (2) and 3-O-(2'E,4'Z)-decadienoylingenol (3), were isolated from Euphorbia kansui under the bioassay-guided method. Each compound showed the same antinematodal activity against the nematode, Bursaphelenchus xylophilus, at a minimum effective dose (MED) of 5 microg/cotton ball. PMID- 17420579 TI - Influence of maceration temperature in red wine vinification on extraction of phenolics from berry skins and seeds of grape (Vitis vinifera). AB - The extraction of phenolics from berry skins and seeds of the grape, Vitis vinifera cv. Cabernet Sauvignon, during red wine maceration and the influence of different temperature conditions (cold soak and/or heating at the end of maceration) were examined. Phenolics contained mainly in berry skins, viz., anthocyanin, flavonol, and epigallocatechin units within proanthocyanidins, were extracted during the early stage of maceration, whereas those in seeds, viz., gallic acid, flavan-3-ol monomers, and epicatechin-gallate units within proanthocyanidins, were gradually extracted. In addition to their localization, the molecular size and composition of the proanthocyanidins possibly influenced the kinetics of their extraction. Cold soak reduced the extraction of phenolics from the seeds. Heating at the end of maceration decreased the concentration of proanthocyanidins. Thus, modification of the temperature condition during maceration affected the progress of the concentration of phenolics, resulting in an alteration of their make-up in the finished wine. PMID- 17420580 TI - Effects of astaxanthin in obese mice fed a high-fat diet. AB - Astaxanthin is a natural antioxidant carotenoid that occurs in a wide variety of living organisms. We investigated the effects of astaxanthin supplementation in obese mice fed a high-fat diet. Astaxanthin inhibited the increases in body weight and weight of adipose tissue that result from feeding a high-fat diet. In addition, astaxanthin reduced liver weight, liver triglyceride, plasma triglyceride, and total cholesterol. These results suggest that astaxanthin might be of value in reducing the likelihood of obesity and metabolic syndrome in affluent societies. PMID- 17420581 TI - Strong enhancement of recombinant cytosine deaminase activity in Bifidobacterium longum for tumor-targeting enzyme/prodrug therapy. AB - In our previous studies, a strain of the nonpathogenic, anaerobic, intestinal bacterium, Bifidobacterium longum (B. longum), was found to be localized selectively and to proliferate within solid tumors after systemic administration. In addition, B. longum transformed with the shuttle-plasmid encoding the cytosine deaminase (CD) gene expressed active CD, which deaminated the prodrug 5 fluorocytosine (5-FC) to the anticancer agent 5-fluorouracil (5-FU). We also reported antitumor efficacy with the same plasmid in several animal experiments. In this study, we constructed a novel shuttle-plasmid, pAV001-HU-eCD-M968, which included the mutant CD gene with a mutation at the active site to increase the enzymatic activity. In addition, the plasmid-transformed B. longum produces mutant CD and strongly increased (by 10-fold) its 5-FC to 5-FU enzymatic activity. The use of B. longum harboring the new shuttle-plasmid increases the effectiveness of our enzyme/prodrug strategy. PMID- 17420582 TI - Novel DPPH radical scavengers, demethylbisorbibutenolide and trichopyrone, from a fungus. AB - In our screening program for antioxidants with 1,1-diphenyl-2-picrylhydrazyl (DPPH)-radical scavenging activity, two novel compounds, demethylbisorbibutenolide (1) and trichopyrone (2), were isolated from the fermentation broth of the fungus of USF-4860 strain isolated from a soil sample. The structures of these compounds were determined from spectroscopic evidence. The biosynthetic origin of the carbon atoms of 2 was unambiguously determined by feeding experiments using (13)C-labeled precursors and elucidation of the (13)C NMR spectrum of (13)C-labeled 2. These studies showed that 2 was derived from five acetates and a methyl group of methionine. In the DPPH-radical scavenging assay, 1 and 2 gave ED(50) values of 149 and 167 muM after standing for 2.0 hr. Compound 2 reacted with the DPPH radical to form reaction product 3 which was determined to be 1-[4-(3,4-dihydro-3-methyl-6-{1,3-pentadienyl}-2,4-dioxo-2H pyran-3-yl)-phenyl]-1-phenyl-2-picrylhydrazine from spectroscopic evidence. PMID- 17420583 TI - Antifeedants against Locusta migratoria from the Japanese Cedar, Cryptomeria japonica. AB - (1S,6R)-2,7(14),10-bisabolatrien-1-ol-4-one and (+)-7(14),10-bisaboladien-1-ol-4 one were isolated and identified from Cryptomeria japonica as antifeedants against Locusta migratoria L. which is well known as a serious pest to cereals throughout the world. These compounds strongly inhibited the feeding of L. migratoria only when they were combined, but each compound alone did not show any activity. PMID- 17420584 TI - The SskA and SrrA response regulators are implicated in oxidative stress responses of hyphae and asexual spores in the phosphorelay signaling network of Aspergillus nidulans. AB - Histidine-to-Aspartate (His-Asp) phosphorelay (or two-component) systems are common signal transduction mechanisms implicated in a wide variety of cellular responses to environmental stimuli in both prokaryotes and eukaryotes. For a model filamentous fungi, Aspergillus nidulans, in this study we first compiled a complete list of His-Asp phosphorelay components, including 15 genes for His kinase (HK), four genes for response regulator (RR), and only one for histidine containing phosphotransfer intermediate (HPt). For these RR genes, a set of deletion mutants was constructed so as to create a null allele for each. When examined these mutant strains under various conditions stressful for hyphal growth and asexual spore development, two of them (designated DeltasskA and DeltasrrA) showed a marked phenotype of hypersensitivity to oxidative stresses (particularly, to hydrogen peroxide). In this respect, expression of the vegetative-stage specific catB catalase gene was severely impaired in both mutants. Furthermore, conidia from DeltasskA were hypersensitive not only to treatment with H(2)O(2), but also to treatment at aberrantly low (4 degrees C) and high (50 degrees C) temperatures, resulting in reduced germination efficiency. In this respect, not only the catA catalase gene specific for asexual development, but also a set of genes encoding the enzymes for synthesis of certain stress tolerant compatible solutes, such as trehalose and glycerol, were markedly downregulated in conidia from DeltasskA. These results together are indicative of the physiological importance of the His-Asp phosphorelay signaling network involving the SskA and SrrA response regulators. PMID- 17420585 TI - Characterization of two biphenyl dioxygenases for biphenyl/PCB degradation in A PCB degrader, Rhodococcus sp. strain RHA1. AB - Rhodococcus sp. RHA1 induces two biphenyl dioxygenases, the BphA and EtbA/EbdA dioxygenases, during growth on biphenyl. Their subunit genes were expressed in R. erythropolis IAM1399 to investigate the involvement of each subunit gene in their activity and their substrate preferences. The recombinant expressing ebdA1A2A3etbA4 and that expressing bphA1A2A3A4 exhibited 4-chlorobiphenyl (4-CB) transformation activity, suggesting that these gene sets are responsible for the EtbA/EbdA and BphA dioxygenases respectively. When bphA4 and etbA4 were swapped to construct the recombinants expressing ebdA1A2A3bphA4 and bphA1A2A3etbA4 respectively, compatibility between BphA4 and EtbA4 was suggested by their 4-CB transformation activities. When bphA3 and ebdA3 were swapped, incompatibility between BphA3 and EbdA3 was suggested. BphA and EtbA/EbdA dioxygenases exhibited the highest transformation activity toward biphenyl and naphthalene respectively, and also attacked dibenzofuran and dibenzo-p-dioxin. The wide substrate preference of EtbA/EbdA dioxygenase suggested that it plays a more important role in polychlorinated biphenyl (PCB) degradation than does BphA dioxygenase. PMID- 17420586 TI - Phylogenetic and disruption analyses of aspartate kinase of Deinococcus radiodurans. AB - The extremely radioresistant bacterium Deinococcus radiodurans is evolutionarily closely related to the extremely thermophilic bacterium Thermus thermophilus. These bacteria have a single gene encoding an aspartate kinase (AK) that catalyzes the phosphorylation of L-aspartate. T. thermophilus has an aminoadipate pathway for lysine biosynthesis that does not use AK for lysine biosynthesis. Phylogenetic analysis in this study indicated that D. radiodurans AK has a different protein structure and a different evolutionary history from T. thermophilus AK. Disruption analysis of D. radiodurans AK indicated that D. radiodurans AK was not used for lysine biosynthesis but for threonine and methionine biosyntheses. A D. radiodurans AK disruption mutant exhibited a phenotype similar to a T. thermophilus AK disruption mutant, which indicates that these two AKs have different evolutionary origins, though their functions are not different. PMID- 17420587 TI - Hypocholesterolemic mechanism of Chlorella: Chlorella and its indigestible fraction enhance hepatic cholesterol catabolism through up-regulation of cholesterol 7alpha-hydroxylase in rats. AB - Chlorella powder (CP) has a hypocholesterolemic effect and high bile acid-binding capacity; however, its effects on hepatic cholesterol metabolism are still unclear. In the present study, male Wistar rats were divided into four groups and fed a high sucrose + 10% lard diet (H), an H + 10% CP diet (H+CP), an H + 0.5% cholesterol + 0.25% sodium cholate diet (C), or a C + 10% CP diet (C+CP) for 2 weeks. CP decreased serum and liver cholesterol levels significantly in rats fed C-based diets, but did not affect these parameters in rats fed H-based diets. CP increased the hepatic mRNA level and activity of cholesterol 7alpha-hydroxylase (CYP7A1). CP increased hepatic HMG-CoA reductase (HMGR) activity in the rats fed H-based diets, but not in rats fed C-based diets. CP did not affect hepatic mRNA levels of sterol 27-hydroxylase, HMGR, low-density lipoprotein (LDL) receptor, scavenger receptor class B1, ATP-binding cassette (ABC) A1, ABCG5, or ABCB11. Furthermore, the effect of a 3.08% Chlorella indigestible fraction (CIF, corresponding to 10% CP) on hepatic cholesterol metabolism was determined using the same animal models. CIF also decreased serum and liver cholesterol levels significantly in rats fed C-based diets. CIF increased hepatic CYP7A1 mRNA levels. These results suggest that the hypocholesterolemic effect of CP involves enhancement of cholesterol catabolism through up-regulation of hepatic CYP7A1 expression and that CIF contributes to the hypocholesterolemic effect. PMID- 17420588 TI - The N-terminal region is important for the nuclease activity and thermostability of the flap endonuclease-1 from Sulfolobus tokodaii. AB - This paper reports the biochemical properties of two types of recombinant flap endonuclease-1 (FEN-1) proteins obtained from the thermophilic crenarchaeon, Sulfolobus tokodaii strain 7. One of the two FEN-1 proteins is a product of the gene with AUG as the translational start codon (StoS-FEN-1), which is originally assigned in the database. The other is a product of the gene with a new AUG start codon (StoL-FEN-1), which is inserted at 153 bases upstream of the original AUG codon. Although StoL-FEN-1 showed activity and thermostability, StoS-FEN-1 showed neither activity nor thermostability. The N-terminal region in StoL-FEN-1 was also conserved in all of the FEN-1 homologs deduced from genes from newly isolated Sulfolobus spp. These results strongly suggest that the actual start codon of the fen-1 gene from S. tokodaii is not the originally assigned AUG, but rather is located at about 100 bases upstream of this codon. PMID- 17420589 TI - A new proteasome inhibitor, TP-110, induces apoptosis in human prostate cancer PC 3 cells. AB - Proteasome inhibitors are useful in the treatment of cancer. Recently, we found a new proteasome inhibitor, TP-110, derived from tyropeptin A produced by Kitasatospora sp. Here we report that TP-110 induces apoptosis in human prostate cancer PC-3 cells. TP-110 showed strong cytotoxicity to PC-3 cells (IC(50)=0.05 muM). It increased the number of cells in the G(2)-M phase and increased the accumulated amounts of the p21 and p27 proteins, which are negative regulators of cell cycle progression. Furthermore, it induced apoptosis along with chromatin condensation and DNA fragmentation in PC-3 cells, and TP-110-induced apoptosis appeared to be associated with caspase activation. Additionally, TP-110 inhibited not only the degradation of IkappaB and the nuclear translocation of nuclear factor-kappaB (NF-kappaB), but also the DNA binding activity of NF-kappaB. These results indicate that TP-110 shows a strong growth inhibition and apoptosis in PC 3 cells. PMID- 17420590 TI - Mode of action of a germination-specific cortex-lytic enzyme, SleC, of Clostridium perfringens S40. AB - The hydrolysis of the bacterial spore peptidoglycan (cortex) is a crucial event in spore germination. It has been suggested that SleC and SleM, which are conserved among clostridia, are to be considered putative cortex-lytic enzymes in Clostridium perfringens. However, little is known about the details of the hydrolytic process by these enzymes during germination, except that SleM functions as a muramidase. Muropeptides derived from SleC-digested decoated spores of a Bacillus subtilis mutant that lacks the enzymes, SleB, YaaH and CwlJ, related to cortex hydrolysis were identified by amino acid analysis and mass spectrometry. The results suggest that SleC is most likely a bifunctional enzyme possessing lytic transglycosylase activity and N-acetylmuramoyl-L-alanine amidase activity confined to cross-linked tetrapeptide-tetrapeptide moieties of the cortex structure. Furthermore, it appears that during germination of Clostridium perfringens spores, SleC causes merely small and local changes in the cortex structure, which are necessary before SleM can function. PMID- 17420591 TI - Ferulic acid supplementation prevents trimethyltin-induced cognitive deficits in mice. AB - This study's objective was to clarify the ameliorative effects ferulic acid (4 hydroxy-3-methoxycinnamic acid) has against cognitive deficits and ChAT activation in trimethyltin (TMT) induced, memory injured mice following a 28-d ferulic acid treatment. After administering TMT for 3 d, each mouse performed Y maze and passive avoidance tests to check immediate working memory performance and cognitive function. The results showed that ferulic acid administration attenuated TMT-induced memory injury and a decline in ChAT activity in the mice. This suggests that ferulic acid might be useful for preventing cognitive dysfunction as well as for boosting the activation of ChAT in dementia. PMID- 17420592 TI - Cloning and expression of a novel sulfotransferase with unique substrate specificity from Bombyx mori. AB - We identified a cDNA encoding a putative cytosolic sulfotransferase (SULT) by searching the expressed sequence tag database of Bombyx mori, and subsequently obtained the full-length cDNA for this gene via rapid amplification of cDNA ends (RACE). We designated this gene bmST1, and showed by sequence analysis that it belongs to a novel SULT family. The tissue specificity of bmST1 mRNA expression was examined in fifth instar larvae by reverse transcriptase-polymerase chain reaction (RT-PCR), and transcripts were detectable in the silk gland, gut, fat body, and Malpighian tube. A recombinant form of bmST1 was then expressed using a gluthathione S-transferase (GST) gene fusion system, and it was purified from Escherichia coli. Purified bmST1 did not exhibit sulfating activity toward SULT substrates such as 4-nitrophenol, vanillin, hydroxysteroids, or monoamines. Surprisingly, however, recombinant bmST1 showed considerable activity toward 4 nitrocatechol and also gallate esters, although the catechins are not sulfated by this enzyme. PMID- 17420593 TI - The beta-1,3-exoglucanase gene exgA (exg1) of Aspergillus oryzae is required to catabolize extracellular glucan, and is induced in growth on a solid surface. AB - The biological role of ExgA (Exg1), a secretory beta-1,3-exoglucanase of Aspergillus oryzae, and the expression pattern of the exgA (exg1) gene were analyzed. The exgA disruptant and the exgA-overexpressing mutant were constructed, and phenotypes of both mutants were compared. Higher mycelial growth rate and conidiation efficiency were observed for the exgA-overexpressing mutant than for the exgA disruptant when beta-1,3-glucan was supplied as sole carbon source. On the other hand, no difference in phenotype was observed between them in the presence or absence of the inhibitors of cell wall beta-glucan remodeling when grown with glucose. exgA Expression was induced in growth on solid surfaces such as filter membrane and onion inner skin. A combination of poor nutrition and mycelial attachment to a hydrophobic solid surface appears to be an inducing factor for exgA expression. These data suggest that ExgA plays a role in beta glucan utilization, but is not much involved in cell wall beta-glucan remodeling. PMID- 17420594 TI - Improvement of digestibility, reduction in allergenicity, and induction of oral tolerance of wheat gliadin by deamidation. AB - Wheat gliadin was deamidated by using a cation-exchange resin in the presence or absence of added cysteine, with the change in digestibility being measured. The allergenicity of the gliadin was evaluated by using sera from patients RAST positive to wheat. Gliadin-specific IgE was measured after the gliadin had been orally administered to rats. The addition of cysteine before the treatment with a cation exchanger effectively increased the deamidation level of gliadin. Deamidated gliadin showed higher solubility than the undeamidated form. There was no difference in the peptic digestibility of the gliadin, whereas deamidation enhanced the pancreatic digestibility in vitro and the digestibility in the mouse stomach in vivo. Deamidation of gliadin reduced its reactivity toward the sera of patients with wheat allergy. Rats administered with deamidated gliadin showed suppressed elevation of the gliadin-specific IgE level. PMID- 17420595 TI - Improvement of dibenzothiophene desulfurization activity by removing the gene overlap in the dsz operon. AB - Dibenzothiophene (DBT) and its derivatives can be microbially desulfurized by Dsz enzymes. We investigated the expressional characteristics of the dsz operon. The result revealed that the ratio of mRNA quantity of dszA, dszB, and dszC was 11:3.3:1; however, western blot analysis indicated that the expression level of dszB is far lower than that of dszC. Gene analysis revealed that the termination codon of dszA and the initiation codon of dszB overlapped, whereas there was a 13 bp gap between dszB and dszC. In order to get a better, steady expression of DszB, we removed this structure by overlap polymerase chain reaction (PCR) and expressed the redesigned dsz operon in Rhodococcus erythropolis. The desulfurization activity of resting cells prepared from R. erythropolis DR-2, which held the redesigned dsz operon, was about five-fold higher than that of R. erythropolis DR-1, which held the original dsz operon. PMID- 17420596 TI - Antifungal activity of tetra-substituted tetrahydrofuran lignan, (-)-virgatusin, and its structure-activity relationship. AB - Antifungal activities of the optically pure (>99%ee) (-)- and (+)-virgatusin, a tetra-substituted tetrahydrofuran lignan, were tested. (-)-Virgatusin, which is a natural product, showed highest antifungal activity against Colletotrichum lagenarium. Research on its structure-activity relationship was also performed. It was shown that two methoxy groups on 9 and 9' positions and a 3,4 methylenedioxyphenyl group on the 7 position of virgatusin were essential for high fungal growth inhibition. The part on 7'-phenyl group was not essential for activity. The 7'-(4-methoxyphenyl) derivative showed higher activity than that of (-)-virgatusin. PMID- 17420597 TI - Suppression of postprandial hypertriglyceridemia in rats and mice by oolong tea polymerized polyphenols. AB - Oolong tea-polymerized polyphenols (OTPP) are characterized polyphenols produced from semi-fermented tea (oolong tea). In the present study, we evaluated the suppressive effects of oolong tea extract and OTPP on postprandial hypertriglyceridemia in rats and mice. Lymphatic recovery of triglycerides in rats cannulated in the thoracic duct was delayed by the administration of oolong tea extract at 100 and 200 mg per head, and more effectively than with green tea extract. OTPP delayed lymphatic triglyceride absorption at 20 mg/head, though (-) epigallocatechin gallate (EGCG) did not do so at the same dose. OTPP also suppressed postprandial hypertriglyceridemia after administration of olive oil in mice. The area under the curve (AUC) of plasma triglycerides was significantly decreased, by 53% and 76%, in the 500 and 1,000 mg/kg OTPP groups respectively, as compared with the control group. These results suggest that OTPP is responsible for the suppression of hypertriglyceridemia by ingestion of oolong tea. PMID- 17420598 TI - Bioconversion of ferulic acid to 4-vinylguaiacol and 4-ethylguaiacol and of 4 vinylguaiacol to 4-ethylguaiacol by halotolerant yeasts belonging to the genus Candida. AB - In order to examine the genesis of the characteristic flavors of soy sauce and miso, seven novel halotolerant yeast strains of two types, which showed convertibility of ferulic acid (FA) to 4-vinylguaiacol (4-VG) and to 4 ethylguaiacol (4-EG), were isolated from miso-koji and miso pastes. Two of these strains were identified as Candida guilliermondii (anamorph of Pichia guilliermondii), and Candida fermentati (anamorph of Pichia caribbica), based on sequence analyses of a partial 26S ribosomal RNA gene and the region of internal transcribed spacers 1 and 2, and the 5.8S ribosomal RNA gene. Moreover, we also found three Candida etchellsii strains which showed convertibility of FA to 4-VG, but not to 4-EG, and two atypical strains of Candida versatilis which showed no convertibility of FA to 4-VG, but did show convertibility of 4-VG to 4-EG from soy sauce mashes. The bioconversion pathway from FA to 4-EG via 4-VG in halotolerant yeasts and bacteria is discussed. PMID- 17420599 TI - Phylogenetic analysis of the gut bacterial microflora of the fungus-growing termite Odontotermes formosanus. AB - We constructed a bacterial 16S rRNA gene clone library from the gut microbial community of O. formosanus and phylogenetically analyzed it in order to contribute to the evolutional study of digestive symbiosis and method development for termite control. After screening by restriction fragment length polymorphism (RFLP) analysis, 56 out of 280 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. The representative phylotypes were affiliated to four phylogenetic groups, Firmicutes, the Bacteroidetes/Chlorobi group, Proteobacteria, and Actinobacteria of the domain Bacteira. No one clone affiliated with the phylum Spirochaetes was identified, in contrast to the case of wood-feeding termites. The phylogenetic analysis revealed that nearly half of the representative clones (25 phylotypes) formed monophyletic clusters with clones obtained from other termite species, especially with the sequences retrieved from fungus-growing termites. These results indicate that the presence of termite-specific bacterial lineages implies a coevolutional relationship of gut microbes and host termites. PMID- 17420600 TI - Induction of apoptosis in DLD-1 human colon cancer cells by peridinin isolated from the dinoflagellate, Heterocapsa triquetra. AB - Peridinin, which is uniquely present in dinoflagellates, is one of the most abundant carotenoids found in nature. We evaluated the apoptotic effect of peridinin on DLD-1 human colorectal cancer cells. Peridinin significantly reduced the cell viability in a dose-dependent manner (0-20 microM) and induced apoptosis by activating both caspase-8 and caspase-9. Our findings could be important for the high-performance utilization of marine bioproducts. PMID- 17420601 TI - Characterization of dnaJ multigene family in the cyanobacterium Synechococcus elongatus PCC 7942. AB - The genome of the cyanobacterium Synechococcus elongatus PCC 7942 contains four dnaJ homologs, which are classified into three types based on domain structure. Among these, dnaJ1, dnaJ2, and dnaJ3 are essential for normal growth, and hence we analyzed them with a view to characterizing their specificity. Expression analysis indicated that dnaJ2, which encodes type II DnaJ protein, exhibited typical responses to heat and high-light stresses. Their localization and ability to prevent aggregation of luciferase were also diverse, suggesting a possible functional differentiation of these proteins. Since the expression of dnaJ1, which belongs to conserved type I DnaJ, down-regulated under heat stress, the unique structure of DnaJ2 may be involved in stress responses of S. elongatus. Based on phylogenetic analysis, the diverse dnaJ family was assumed to have evolved its own specific functions in each cyanobacterial species. PMID- 17420602 TI - Kinetic analysis of glucoamylase-catalyzed hydrolysis of starch granules from various botanical sources. AB - The kinetics of glucoamylase-catalyzed hydrolysis of starch granules from six different botanical sources (rice, wheat, maize, cassava, sweet potato, and potato) was studied by the use of an electrochemical glucose sensor. A higher rate of hydrolysis was obtained as a smaller size of starch granules was used. The adsorbed amount of glucoamylase on the granule surface per unit area did not vary very much with the type of starch granules examined, while the catalytic constants of the adsorbed enzyme (k(0)) were determined to be 23.3+/-4.4, 14.8+/ 6.0, 6.2+/-1.8, 7.1+/-4.1, 4.6+/-3.0, and 1.6+/-0.6 s(-1) for rice, wheat, maize, cassava, sweet potato, and potato respectively, showing that k(0) was largely influenced by the type of starch granules. A comparison of the k(0)-values in relation to the crystalline structure of the starch granules suggested that k(0) increases as the crystalline structure becomes dense. PMID- 17420603 TI - Prevention of pathogenic Escherichia coli infection in mice and stimulation of macrophage activation in rats by an oral administration of probiotic Lactobacillus casei I-5. AB - Lactobacillus casei I-5 isolated from an alcohol fermentation broth enhanced immunity and prevented pathogenic infection as a probiotic. Mice fed with I-5 cells for 11 days prior to an intraperitoneal challenge with pathogenic Escherichia coli Juhl exhibited a high survival rate compared with the control group. Rats fed with I-5 cells for 10 days significantly increased the phagocytosis of peritoneal macrophages. In a cell culture system employing peritoneal macrophages from rats, the I-5 administration activated NF-kappaB stimulated by LPS. It also enhanced LPS-stimulated IL-12 and TNF-alpha production, but not IL-6 production. These results show that L. casei I-5 effectively prevented infection by pathogenic E. coli possibly through the activation of peritoneal macrophages. The strain would be useful to prevent pathogenic microbial infections in humans and farm animals. PMID- 17420604 TI - Effect of Kluyveromyces marxianus YIT 8292 crude cell wall fraction on serum lipids in normocholesterolemic and mildly hypercholesterolemic subjects. AB - The hypocholesterolemic effects of Kluyveromyces marxianus YIT 8292 crude cell wall (KM-CW) were examined. In pilot studies, KM-CW tablets were administered to mildly hypercholesterolemic subjects at doses of 8.0, 4.0, 2.0, or 1.0 g/d for 4 weeks. Total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) decreased at doses above 2.0 and 4.0 g/d, respectively. Further, we examined the effect of intake of yogurt containing 3.0 or 4.0 g of KM-CW/d for 8 weeks in normal and hypercholesterolemic subjects in a double-blind placebo-controlled study. The intake of either of the KM-CW-containing yogurts was associated with significantly improved TC and LDL-C in hypercholesterolemic subjects, but had no effect on these levels in normal subjects. TC was significantly lower at week 8 in the hypercholesterolemic subjects who ingested yogurt containing 3.0 or 4.0 g of KM-CW than in those who consumed placebo yogurt. Intake of KM-CW might contribute to the prevention of hypercholesterolemia. PMID- 17420605 TI - Hepatoprotective effects of a concentrate and components of sake against galactosamine (GalN)-induced liver injury in mice. AB - We investigated the hepatoprotective effects of a concentrate of sake (CS) and its components against D-galactosamine (GalN)-induced liver injury by measuring the plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in mice. CS significantly suppressed the GalN-induced elevation of ALT and AST activities. Each of four concentrated fractions extracted from sake (respectively consisting mainly of basic amino acids, neutral and acidic amino acids, organic acids and sugars) suppressed the GalN-induced elevation of ALT and AST activities. We focused on the sugar fraction containing glucose and ethyl alpha-D-glucoside (alpha-EG), which is a sake-specific sugar, as the major components and demonstrated that only alpha-EG showed significant suppression of the GalN-induced elevation of ALT and AST activities. We compared the effects of the alpha-EG analogues, methyl alpha-D-glucoside and ethyl beta-D-glucoside, on GalN-induced liver injury and confirmed that only alpha-EG significantly suppressed both the ALT and AST activities. Moreover, CS and alpha-EG suppressed the GalN-induced production of interleukin 6 (IL-6) and liver DNA fragmentation. Together these results show that CS and its component, alpha-EG, suppressed GalN induced liver injury by inhibiting IL-6 production. PMID- 17420606 TI - Isolation and characterization of a spore germination inhibitor from Streptomyces sp. CB-1-1, a phytopathogen causing root tumor of melon. AB - The germination rate and activation conditions of spores were examined for four strains of Streptomyces sp., a phytopathogen causing root tumor of melon. An inhibitor was isolated from the agar-cultured material of strain CB-1-1 and then characterized. The inhibitor selectively acted on spore germination and did not affect hyphal growth, and inhibition was abolished by washing the spores in water. The inhibitor was produced by an agar culture, and most of the inhibitor existed in the spores. The IC(50) value for the inhibitor was approximately 0.25 microg/ml. PMID- 17420607 TI - Zerumbone suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells. AB - Unregulated uptake of oxidized low-density lipoproteins (ox-LDL) via macrophage scavenger receptors (SRs), such as lectin-like ox-LDL receptor-1 (LOX-1), is a key event in atherosclerosis. In the present study, we used differentiated Caco-2 cells as a model of the human small intestine to evaluate the suppressive effects of 16 traditional food items selected from Okinawa on 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced LOX-1 mRNA expression in THP-1 human monocyte-like cells. Three Zingiberaceae plants, Curcuma aromatica Salisbury, Curcuma longa L., and Zingiber zerumbet Smith, markedly suppressed that expression. When added to the apical sides of Caco-2 monolayers, zerumbone, a sesquiterpene from Z. zerumbet Smith, was found to permeate into the basolateral medium as an intact structure in a time-dependent manner. alpha-Humulene, a structural analog of zerumbone lacking the alpha,beta-unsaturated carbonyl group, did not suppress LOX 1 mRNA expression, indicating that its electrophilic moiety might play pivotal roles in its activities. Further, zerumbone attenuated the expression of SR-A, SR PSOX, and CD36, but not that of CD68 or CLA-1, leading to a blockade of DiI-acLDL uptake, while it also inhibited the transcriptional activities of activator protein-1 and nuclear factor-kappaB. Together, our results indicate that zerumbone is a potential phytochemical for regulating atherosclerosis with reasonable action mechanisms. PMID- 17420608 TI - Marked improvement of psychiatric symptoms after parathyroidectomy in elderly primary hyperparathyroidism. AB - Psychosomatic symptoms in primary hyperparathyroidism (PHPT) are various and include such conditions as obsessive-compulsive disorder, depression, anxiety, and paranoia. In the elderly the clinical features of the disease are often non specific and difficult to diagnose. To quantify subjective symptoms of patients with hyperparathyroidism in the elderly, we determined whether these clinical manifestations resolved after surgical parathyroidectomy (PTX) in three PHPT patients over eighty years old. They were diagnosed with hypercalcemia, hypophosphatemia, high PTH concentrations, and osteoporosis. A single parathyroid adenoma was confirmed in each patient by Tc-MIBI scintigram, neck ultrasonography and computed tomographic scanning. PTX was performed in these three patients. Assessments of psychologic symptoms, using the Hamilton Rating Scale for Depression (HAM-D), serum calcium, and intact PTH were obtained before and after PTX. Mean weight of the resected adenomas was 438 +/- 138 mg (mean +/- SD). After PTX, serum calcium decreased from 11.1 +/- 0.5 to 9.2 +/- 0.5 mg/dl and intact PTH from 160.0 +/- 25.2 to 45.3 +/- 22.2 pg/ml. Total HAM-D scores in each patient decreased from 45 to 9, 17 to 1 and 15 to 5, respectively. Especially, there were marked improvements in depressive mood, psychomotor inhibition, anxiety and somatic symptoms after PTX. The quality of life in those patients was also improved by PTX. We propose here that PTX in elderly PHPT patients with psychiatric symptoms should be considered instead of oral administration, such as anti-depressants or bisphosphonates. PMID- 17420609 TI - Somatostatin receptor subtypes mRNA in TSH-secreting pituitary adenomas: a case showing a dramatic reduction in tumor size during short octreotide treatment. AB - TSH-secreting adenoma is a rare pituitary adenoma, and the expression levels of the specific subtypes of somatostatin receptors (sstr) mRNAs have remained obscure. To determine the quantitative expression of the sstr1-5 mRNAs in TSH secreting adenomas that may be related to the efficacy of treatment with a somatostatin analogue, expression of the sstr1-5 mRNAs was examined and compared in TSH-secreting adenomas and other pituitary adenomas. The pituitary adenomas were obtained at transsphenoidal surgery from 4 cases of TSH-secreting adenoma, including 1 patient showing a significant shrinkage of the tumor size after only 10 days of octreotide treatment, 2 patients without tumor size reduction and 1 patient without treatment, and 5 GH-secreting adenomas, 6 prolactinomas, 5 nonfunctioning adenomas, 4 ACTH-secreting adenomas and normal pituitaries at autopsy from 4 normal subjects. In comparison to the normal pituitary, sstr2A>sstr1>sstr5>sstr3 mRNAs were expressed in the TSH-secreting adenomas examined. No expression of sstr2B or sstr4 mRNA was observed. The expression level of sstr2 mRNA was significantly higher than those in normal pituitary, prolactinomas, ACTH-secreting and nonfunctioning pituitary adenomas. The patient with marked shrinkage of the tumor showed the highest expression of both sstr2 and sstr5 mRNAs among all the cases of pituitary adenoma. A TSH-secreting tumor without shrinkage showed a similar expression level of sstr2 mRNA. These findings demonstrated that TSH-secreting adenomas express sstr1, 2A, 3 and 5 mRNAs, predominantly sstr2A, and in addition to the expression of sstr2 mRNA, the expression level of sstr5 mRNA may be a factor affecting the tumor shrinkage by somatostatin analogues against TSH-secreting adenomas. PMID- 17420610 TI - Life expectancy among Japanese of different smoking status in Japan: NIPPON DATA80. AB - BACKGROUND: The life expectancy is an important measure for describing health status among population. Several studies from the United States and Europe showed the harm of smoking by describing the life expectancies with different smoking status. No such study is examined in Japan, the country with the world's highest life expectancy irrespective of high smoking rate among men. METHODS: The abridged life table method was applied to calculate the life expectancies of men and women among different smoking status from age 40 until age 85. Age-specific mortality rates stratified by different smoking status were obtained from follow up data from random sample in Japanese population (NIPPON DATA80). RESULTS: Proportion of current smokers was 62.9% in men and 8.8% in women at the baseline survey in 1980. The life expectancies of 40-year-old never smokers, ex-smokers and current smokers were 42.1, 40.4, and 38.6 years in men and 45.6, 45.9, and 43.4 years in women. The life expectancy of 40-year-old men who smoked less than one pack per day was 39.0 and was longer than that of those who smoked one or two packs (38.8) and more than two packs (38.1). CONCLUSION: Life expectancy decreased gradually as the grade of smoking increased in the Japanese population. PMID- 17420611 TI - Consumption of dairy products and cancer risks. AB - BACKGROUND: Relationships between consumption of dairy products and death from various types of cancer are largely unknown. METHODS: Between April 1992 and July 1995, a baseline survey was conducted for 11,349 residents in 12 communities in Japan, which included collection of demographic data and a self-administered food frequency questionnaire inquiring about three dairy products: milk, butter and yogurt. The subjects were followed prospectively until 2002. Causes of death were identified using death certificates. Hazard ratios (HRs) and their 95% confidence intervals (CIs) for each dairy product were calculated using Cox's proportional hazard models. RESULTS: Among eight common cancers, only deaths from hematopoietic neoplasm (n=14) were significantly associated with consumption of butter (HR=5.11, 95% CI: 1.40-18.62), though they exhibited a nearly-significant association with milk consumption (HR=3.17, 95% CI: 0.99-10.17), independent of age and sex. Consumption of milk and butter was significantly associated with non lymphoma deaths (n=9) when adjusted for age and sex (HR=9.86, 95% CI: 1.23-79.19 for milk; and HR=10.04, 95% CI: 2.39-42.18 for butter). CONCLUSION: The frequencies of butter consumption, and probably that of milk, were correlated with death from hematopoietic neoplasm, particularly from non-lymphomas. PMID- 17420612 TI - Calcium intake pattern among Japanese women across five stages of health behavior change. AB - BACKGROUND: The transtheoretical model (TTM) of health behavior change is one of the most promising approaches for health professionals to help individuals change their behaviors. Few studies have assessed calcium intake using the model on Asian women. This study aims at clarifying characteristics of each behavioral stage among Japanese women and providing clues to increase calcium intake to prevent osteoporosis. METHODS: A cross-sectional survey was conducted from September through November, 2005 using self-administered questionnaires. A total of 226 participants in an osteoporosis screening program were invited to take part in the study, and 150 women were enrolled. RESULTS: Adjusted means of total dietary calcium were positively significantly associated with successive stages (p<0.001). The proportion of calcium intake from plants and fish was higher in the precontemplation, contemplation, and preparation stages compared with the action and maintenance stages (p=0.038). Concomitantly, the plants and fish food group contributed 46.7% of total dietary calcium, while 32.4% was derived from milk and dairy food, and 20.9% from other foods. The correlation coefficient (95% confidence interval) between the proportion of calcium obtained from plants and fish and the proportion of fat energy was -0.22 (-0.37, -0.06). CONCLUSIONS: The proportion of calcium intake from plants and fish was higher among women in the lower stages compared with higher stages. Given the higher prevalence of lactose intolerance, it would seem plausible to recommend lower-stage women be educated and encouraged to derive more calcium from plants and fish diets as a means to prevent osteoporosis. PMID- 17420613 TI - Incidence of chronic obstructive pulmonary disease, and the relationship between age and smoking in a Japanese population. AB - BACKGROUND: Accurately evaluating a risk of chronic obstructive pulmonary disease (COPD) requires a large-scale longitudinal study using a standard criterion for diagnosing COPD. There have been only a few such follow-up studies in Europe and no reports in Asia. We estimated the incidence rate and incidence rate ratio (IRR) of age and smoking for COPD in a Japanese population using the diagnosis criterion of the Global Initiative for Chronic Obstructive Lung Disease guidelines. METHODS: Subjects were 17,106 participants aged 25-74 years during health check-ups including spirometry from April 1997 through March 2005 in Japan. Total follow-up of participants were 47,652 person-years in males and 25,224 person-years in females. The IRR of age and smoking was estimated using Cox proportional hazard models with both variables. RESULTS: We identified 466 incidence cases of COPD. The incidence rate per 100 person-years was 0.81 (95% confidence interval [CI], 0.73-0.89) in males and 0.31 (0.24-0.38) in females, and significantly increased with age in both sexes. The incidence rate for current smokers was significantly higher than that for male non-smokers but not significantly for females. Among males, the IRR for current smokers with Brinkman Index < 400, 400-799, and 800+ was 1.2 (0.8-1.9), 2.7 (1.9-3.8), and 4.6 (3.3 6.5), respectively. CONCLUSION: These results indicated that the COPD risk gradually increased with aging, and that there was a dose-response relationship between smoking and COPD risk. PMID- 17420614 TI - Differential effects of power rehabilitation on physical performance and higher level functional capacity among community-dwelling older adults with a slight degree of frailty. AB - BACKGROUND: Evidence is still insufficient regarding the effects of Power Rehabilitation (PR) on physical performance and higher-level functional capacity of community-dwelling frail elderly people. METHODS: This nonrandomized controlled interventional trial consisted of 46 community-dwelling elderly individuals with light levels of long-term care needs. They were allocated to the intervention (I-group, n = 24) and control (C-group, n = 22) groups. Of them, 32 persons (17 in the I-group; 15 in the C-group) (median age, 77 years; sex, 28% male) completed the study. The I-group subjects underwent PR twice a week for 12 weeks. The outcomes were physical performance (muscle strength, balance, flexibility, and mobility) and higher-level functional capacity as evaluated by the Tokyo Metropolitan Institute of Gerontology Index of Competence (TMIG-IC) and the level of long-term care need as certified by the public long-term care insurance. RESULTS: The I-group demonstrated a significant improvement in the measured value of the timed up-and-go test (median change, a decrease of 4.4 seconds versus a decrease of 0.2 seconds, p = 0.033) and the timed 10-meter walk (a decrease of 3.0 seconds versus an increase of 0.2 seconds, p = 0.007) in comparison with the C-group. No significant change was observed in the TMIG-IC scores or in the level of long-term care need in the I-group. CONCLUSION: PR improved mobility of community-dwelling frail elderly people; however, such improvement did not translate into higher-level functional capacity. Our findings demonstrate the difficulty in transferring the positive effects associated with PR into an improvement in higher-level functional capacity. PMID- 17420615 TI - Membrane stretch-induced activation of a TRPM4-like nonselective cation channel in cerebral artery myocytes. AB - Stretch-activated cation channels (SACs) have been observed in many types of smooth muscle cells. However, the molecular identity and activation mechanisms of SACs remain poorly understood. We report that TRPM4-like cation channels are activated by membrane stretch in rat cerebral artery myocytes (CAMs). Negative pressure (> or =20 mmHg, cell-attached mode) activated single channels (approximately 20 pS) in isolated CAMs. These channels were permeable to Na(+) and Cs(+) and inhibited by Gd(3+) (30 microM) and DIDS (100 microM). The effect of negative pressure was abolished by membrane excision, but subsequent application of Ca(2+) (>100 nM) to the intracellular side of the membrane restored single channel activity that was indistinguishable from SACs. Caffeine (5 mM), which depletes SR Ca(2+)-stores, first activated and then abolished SACs. Tetracaine (100 microM), a ryanodine receptor antagonist, inhibited SACs. Overexpression of hTRPM4B in HEK293 cells resulted in the appearance of cation channels that were activated by both negative pressure and Ca(2+) and which had very similar biophysical and pharmacological properties as compared with SACs in CAMs. These studies indicate that TRPM4-like channels in CAMs can be activated by membrane stretch, possibly through ryanodine receptor activation, and this may contribute to the depolarization and concomitant vasoconstriction of intact cerebral arteries following mechanical stimulation. PMID- 17420616 TI - Effects of vertically transferred 3,3',4,4',5-pentachlorobiphenyl on gene expression in the ovaries of immature Sprague-Dawley rats. AB - We have previously shown that 3,3',4,4',5-pentachlorobiphenyl (PCB-126) vertically transferred from dams potentially exerts a direct effect on the ovaries of offspring and adversely affects female puberty. To investigate its toxicological targets in ovarian tissues, mRNAs encoding representative peptides that regulate follicular development in granulosa cells, theca cells, and oocytes were quantified using ovaries collected on postnatal days (PND) 5, 15, and 24 from the offspring of dams administered oral doses of 0, 1 or 3 microg/kg PCB-126 starting 2 weeks prior to mating and continuing until 20 days after delivery. Quantification using the real-time RT-PCR method revealed that PCB-126 lowered the amounts of mRNAs that encoded the inhibin alpha- and inhibin/activin beta A subunits from PND 15 onwards; the amounts of mRNAs for inhibin/activin beta B subunit, follicle-stimulating hormone (FSH) receptor, and aromatase on PND 15; and the amounts of luteinizing hormone receptor mRNA on PND 24 compared with those of the age-matched controls. In contrast, no differences were noted for mRNAs encoding c-kit, growth differentiation factor-9, bone morphogenetic protein 15, or kit ligand for any of the age groups examined. The serum FSH level on PND 24 was higher than that in the control. Since the earliest effects on the mRNAs in the rat ovaries were observed in those expressed in the granulosa cells of the growing follicles after the antral follicles had developed, molecules in granulosa cells but not in oocytes during the early stages of the antral follicles might be the primary targets of vertically transferred PCB-126. PMID- 17420617 TI - Reliability of estrous detection in Holstein heifers using a radiotelemetric pedometer located on the neck or legs under different rearing conditions. AB - To evaluate the efficiency and accuracy of estrous detection using a new pedometry system that can measure the hourly activity of cattle, pedometers were attached to the neck and the hind legs of 15 Holstein heifers. Heifers were reared in pasture for grazing, an open paddock, or in a tie-stall barn (an additional pedometer was attached to a front leg of each of these heifers). The most recent 24 h-total number of steps was compared for each 1 h-interval with the mean value of the preceding days during the reference period (RP). The neck pedometer detected all 10 instances of estrous activity (100%) for the grazing heifers at 1.3 times the thresholds value for a 5-day RP but with only 32% accuracy. The hind leg pedometer, however, obtained 100% efficiency and 83% accuracy at 1.4 times the threshold value for a 7-day RP. The efficiencies and accuracies in detecting 12 instances of estrous activity under the paddock condition were 92 and 65% (neck, 1.3-fold, 7-day RP) and 92 and 100% (hind leg, 1.6- or 1.7-fold, 7-day RP), respectively. Under the tie stall condition, the neck pedometers detected 92% of 23 instances of estrous activity with 34% accuracy (1.2-fold, 3-day RP), and the efficiencies and accuracies of the leg pedometers were 78 and 78% (hind leg, 1.4-fold, 4- or 6-day RP) and 87 and 83% (front leg, 1.4-fold, 7-day RP), respectively. Prediction of ovulation time was more precisely with the leg pedometers than with those under the tie stall conditions. Our preliminary results indicate that this new pedometer system has practical value for estrous detection in heifers under different rearing conditions, which affect the criteria required for detection. Furthermore, they also indicate that a leg pedometer can reliably detect estrus and that a neck pedometer may only be capable of detecting estrus under paddock rearing conditions. PMID- 17420618 TI - Preventive effect of zinc against cadmium-induced oxidative stress in the rat testis. AB - The aim of this study was to investigate the antioxidant role of zinc (Zn) in the Cd-exposed testes of Wistar rats. Subchronic exposure to Cd (CdCl(2), 40 mg/l, per os) for 30 days resulted in a significant reduction in growth rate (-11%) and relative weights of testes (-36%) and seminal vesicles (-80%). Treated rats displayed a decrease in testicular and plasma testosterone levels, respectively ( 70%, P<0.05; -48%, P<0.05), epididymal sperm count (-22%, P<0.05), and spermatozoa motility (-35%, P<0.05). In contrast, Cd increased the malondialdehyde (+46%, P<0.05), metallothionein (+200%, P<0.05), and 8-oxodGuo concentrations (+71%, P<0.05) in the testis. In the gonad, Cd decreased the GPx ( 30%, P<0.05), CAT (-32%, P<0.05), mitochondrial Mn-SOD (-34%, P<0.05), and cytosolic CuZn-SOD (-32%, P<0.05) activities. Zinc supplementation (ZnCl(2), 40 mg/l, per os) in the Cd-exposed rats restored the activities of GPx, CuZn-SOD, and Mn-SOD in the testes to the levels of the control group. Moreover, zinc administration was capable of reducing the elevated levels of malondialdehyde in the testis. Interestingly, zinc supplementation attenuated DNA oxidation induced by Cd in the gonad and restored the testosterone level and sperm count to the levels of the control group. Zinc administration minimized oxidative damage and reversed the impairment of spermatogenesis and testosterone production induced by Cd in the rat testis. PMID- 17420619 TI - Effect of cryoprotectant composition on in vitro viability of in vitro fertilized and cloned bovine embryos following vitrification and in-straw dilution. AB - In this study the efficacy of the combination of glycerol (GLY) and ethylene glycol (EG) as cryoprotectants in a vitrification method developed for direct embryo transfer was evaluated by in vitro development of in vitro fertilized (IVF) and somatic cell nuclear transfer (SCNT) embryos after vitrification. The IVF and SCNT blastocysts were vitrified in either 40% GLY, 30% GLY + 10% EG, or 20% GLY + 20% EG using French straws. After warming, the straws were held vertically for 1 min without shaking and were then placed horizontally for 5 min to dilute the cryoprotectants. After washing, the vitrified-warmed embryos were cultured in vitro for 72 h. There were no differences among the vitrification solutions with respect to the rates of vitrified-warmed IVF and SCNT embryos surviving and developing to the hatched blastocyst stage. However, the rates of development to the hatched blastocyst stage of the SCNT embryos vitrified with 40% GLY tended to be higher than those vitrified with 30% GLY + 10% EG or 20% GLY + 20% EG (26% vs. 7-8%, respectively). The development rates to the hatched blastocyst stage of the IVF and SCNT embryos vitrified with solution containing EG were significantly lower (P<0.05) than those of non-vitrified embryos. These results suggest that use of the combination of GLY and EG as cryoprotectants had no beneficial effect on the viability of embryos after in-straw dilution. However, this method is so simple that it can be used for practical direct transfer of vitrified embryos in the field. PMID- 17420620 TI - Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos. AB - In the present study, we investigated the effect of activation treatments on the actin filament distribution and in vitro development of somatic cell nuclear transfer (SCNT) embryos in miniature pigs. We combined three activation methods, ionomycin (ION), electrical stimulation (ES), and cycloheximide treatment (CH), to prepare seven activation treatments (ION, ES, CH, ION + CH, ION + ES, ES + CH and ION + ES + CH). First, we investigated the activation rate of oocytes and in vitro development of parthenotes. The activation rates of the oocytes in the ION, ES, CH, ION + CH, ION + ES, ES + CH, and ION + ES + CH groups were 42.9, 51.3, 0.0, 82.1, 80.6, 78.1 and 78.6%, respectively, showing that the rates of the combined treatment groups were significantly higher (P<0.05) than those of the single treatment groups. Although there were no significant differences in the activation rates of the combined treatment groups, the developmental rate to blastocysts in the ION + CH treatment group (36.1%) was significantly higher (P<0.05) than the other combined treatment groups (14.6-24.7%). Subsequently, we investigated the in vitro development and distribution of microfilaments in SCNT embryos. The developmental rate to blastcysts of the SCNT embryos in the ION + CH treatment group (11.3%) was significantly higher (P<0.05) than in the ES and ION + ES + CH treatment groups (4.5 and 5.2%, respectively). The rate of normal actin filament distribution in the SCNT embryos activated with ION + CH was significantly higher (P<0.05) than those activated with ES or ION + ES + CH treatment (63.3 vs. 46.8 or 46.4%). In addition, the fragmentation rate of the SCNT embryos activated with ION + CH was significantly lower (P<0.05) than those activated with ION + ES + CH (14.9 vs. 26.1%). The present results suggest that an activation treatment of ionomycin combined with cycloheximide may avoid physical damage to microfilaments and result in improved subsequent development of miniature pig SCNT embryos. PMID- 17420621 TI - Effects of iron and copper in culture medium on bovine oocyte maturation, preimplantation embryo development, and apoptosis of blastocysts in vitro. AB - The aim of this study was to investigate the effects of iron and copper on bovine oocyte maturation, preimplantation embryo development and apoptosis of blastocysts. The concentrations of iron in the culture media were 0 (control), 0.45, 0.81, 1.96 and 3.26 mg/l, and the concentrations of copper were 0 (control), 0.093, 0.27, 0.46 and 0.68 mg/l. The changes in the iron (1.96 mg/l) and copper concentrations (0.46 mg/l) in the culture media were measured after oocyte maturation for 22 h and after zygote culture for 48, 96, 144 and 192 h. The results showed that there were no significant differences in oocyte maturation and cleavage between media containing iron and the control, but the media containing iron had higher (P>0.05) rates of 8-cell embryos, morulae, and blastocysts than the control, and addition of 1.96 mg/l of iron increased the blastocyst rate (P>0.05). The effects of copper on oocyte maturation and cleavage were similar to iron, and addition of 0.46 and 0.68 mg/l of copper increased the rates of morulae and blastocysts (P>0.05). Addition of iron or copper significantly decreased the number of apoptotic blastomeres compared with the control (P>0.05). After oocyte maturation for 22 h and zygote culture for 48 h, the iron concentrations decreased by 3.6 and 9.2%, respectively, and the copper concentrations decreased by 6.5 and 10.9%, respectively. After zygote culture for 96, 144 and 192 h, the iron concentrations decreased by 21.4, 25.5 and 27.0%, respectively, the copper concentrations decreased by 23.9, 28.3 and 30.4%, respectively. In conclusion, iron and copper played an important role in the success of culture of 8-cell embryos, morulae, and blastocysts, and long-term lack of iron or copper increased the number of apoptotic blastomeres. Furthermore, transition of primary demand for trace amounts of iron or copper from the cytoplast to culture medium for utilization by zygotes may occur after in vitro zygote culture for 48 h. PMID- 17420622 TI - Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin. AB - The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk. PMID- 17420623 TI - Radiosensitization by the combination of SR-2508 and paclitaxel in hypoxic human tumor cells in vitro. AB - The two radiosensitizers SR-2508 (etanidazole) and paclitaxel (taxol) have different dose-limiting toxicities in humans. Combination of the two radiosensitizers may increase radiosensitization without increasing toxicity. This study was carried out to determine the synergistic radiosensitizing effect of combination of SR-2508 and paclitaxel in two hypoxic human tumor cell lines: a breast carcinoma (MCF-7) and a carcinoma cervicis (HeLa). The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was used to determine the number of surviving cells. Cell cycle was evaluated by flow cytometry. Cell viability was measured by the ability of single cells to form colonies in vitro. Our data demonstrated that the radiosensitization produced by the two radiosensitizers was additive in hypoxic HeLa cells while held in the G(1) phase of the cell cycle. On the other hand, there was no synergistic radiosensitizing effect in hypoxic MCF-7 cells by combination of the two drugs. Our results suggested that the synergistic radiosensitizing effect of SR-2508 and paclitaxel may be tumor-dependent and that breast cancer may not be a good candidate. This study may provide a new combination of radiosensitizers in radiotherapy for cervical carcinoma. PMID- 17420624 TI - Ibn Sina (Avicenna): the prince of physicians. PMID- 17420625 TI - Review of the current status of intra-arterial thrombolysis for treating acute cerebral infarction: a retrospective analysis of the data from multiple centers in Korea. AB - OBJECTIVE: The purpose of the study was to review the current status of intra arterial (IA) thrombolysis in Korea by conducting a retrospective analysis of the data from multiple domestic centers. MATERIALS AND METHODS: The radiologists at each participating institution were asked to fill out case report forms on all patients who had undergone IA recanalization due to acute anterior circulation ischemia. These forms included clinical, imaging and procedure-related information. A central reader analyzed the CT/MR and angiographic results. The rates of successful recanalization, hemorrhagic transformation and functional outcome were obtained. The univariate analyses were performed together with the multivariate analysis. RESULTS: We analyzed the data from 163 patients, and they had been treated at seven institutes. The initial imaging modalities were CT for 46 patients (28%), MR for 63 (39%), and both for 54 (33%). Various mechanical treatment methods were applied together in 50% of the patients. Radiologically significant hemorrhage was noted in 20/155 patients (13%). We found various factors that influenced the recanalization rate and the occurrence of significant hemorrhagic transformations. The favorable outcome rate, reported as modified Rankin Scale < or = 2, was 40%, and the mortality rate was 11%. The factors that predicted a poor functional outcome were old age (p = 0.01), initially severe neurological symptoms (p < 0.0001), MR findings of a wide distribution of lesions (p = 0.001), involvement of the basal ganglia (p = 0.01), performance of procedures after working hours (p = 0.01), failure of recanalization (p = 0.003), contrast extravasation after the procedure (p = 0.007) and significant hemorrhagic transformation (p = 0.002). The subsequent multivariate analysis failed to show any statistically significant variable. CONCLUSION: There was a trend toward increased dependency on MR imaging during the initial evaluation and increased usage of combined pharmacologic/mechanical thrombolysis. The imaging and clinical outcome results of this study were comparable to those of the previous major thrombolytic trials. PMID- 17420626 TI - Early and delayed myocardial enhancement in myocardial infarction using two-phase contrast-enhanced multidetector-row CT. AB - OBJECTIVE: The purpose of this study was to describe the myocardial enhancement patterns in patients with myocardial infarction using two-phase contrast-enhanced multidetector-row computed tomography (MDCT). MATERIALS AND METHODS: Twenty-three patients with clinically proven myocardial infarction (17 acute myocardial infarction [AMI] and 6 chronic myocardial infarction [CMI]) were examined with two-phase contrast-enhanced ECG-gated MDCT. The presence, location, and patterns of myocardial enhancement on two-phase MDCT images were compared with infarcted myocardial territories determined by using electrocardiogram, echocardiography, thallium-201 single photon emission computed tomography, catheter and MDCT coronary angiography. RESULTS: After clinical assessment, the presence of myocardial infarctions were found in 27 territories (19 AMI and 8 CMI) of 23 patients. Early perfusion defects were observed in 30 territories of all 23 patients. Three territories not corresponding to a myocardial infarction were detected in three patients with AMI and were associated with artifacts. Fourteen of perfusion defects were in the left anterior descending artery territory, four in the left circumflex artery territory, and nine in the right coronary artery territory. Delayed enhancement was observed in 25 territories (17 AMI and 8 CMI) of 21 patients. Delayed enhancement patterns were variable. Transmural early perfusion defects (n =12) were closely associated with transmural late enhancement (n = 5) and subendocardial residual defect with subepicardial late enhancement (n = 5). CONCLUSION: Myocardial infarction showed early perfusion defects and variable delayed enhancement patterns on two-phase contrast-enhanced MDCT. Delayed enhancement technique of MDCT could provide additional information of the location and extent of infarcted myocardium, and could be useful to plan appropriate therapeutic strategies in patients with AMI. PMID- 17420627 TI - Pure and mixed tubular carcinoma of the breast: mammographic and sonographic differential features. AB - OBJECTIVE: We wanted to evaluate the mammographic and sonographic differential features between pure (PT) and mixed tubular carcinoma (MT) of the breast. MATERIALS AND METHODS: Between January 1998 and May 2004, 17 PTs and 14 MTs were pathologically confirmed at our institution. The preoperative mammography (n = 26) and sonography (n = 28) were analyzed by three radiologists according to BI RADS. RESULTS: On mammography, a mass was not detected in eight patients with PT and in one patient with MT (57% vs. 8%, respectively, p = 0.021), which was statistically different. The other findings on mammography and sonography showed no statistical differences between the PT and MT, although the numerical values were different. When the lesions were detected mammographically, an irregularly shaped mass with a spiculated margin was more frequently found in the MT than in the PT (100% vs. 83%, respectively, p = 0.353). On sonography, all 28 patients presented with a mass and most lesions showed as not being circumscribed, hypoechoic masses with an echogenic halo. Surrounding tissue changes and posterior shadowing were more frequently found in the MT than in the PT (75% vs. 50%, respectively, p = 0.253, 58% vs. 19%, respectively, p = 1.000). An oval shaped mass was more frequently found in the PT than in the MT (44% vs. 25%, respectively; p = 0.434). CONCLUSION: PT and MT cannot be precisely differentiated on mammography and sonography. However, the absence of a mass on mammography or the presence of an oval shaped mass would favor the diagnosis of PT. An irregularly shaped mass with surrounding tissue change and posterior shadowing on sonography would favor the diagnosis of MT and also a less favorable prognosis. PMID- 17420628 TI - Local recurrence of hepatocellular carcinoma after segmental transarterial chemoembolization: risk estimates based on multiple prognostic factors. AB - OBJECTIVE: To determine the prognostic factors for local recurrence of nodular hepatocellular carcinoma after segmental transarterial chemoembolization. MATERIALS AND METHODS: Seventy-four nodular hepatocellular carcinoma tumors < or = 5 cm were retrospectively analyzed for local recurrence after segmental transarterial chemoembolization using follow-up CT images (median follow-up of 17 months, 4-77 months in range). The tumors were divided into four groups (IA, IB, IIA, and IIB) according to whether the one-month follow-up CT imaging, after segmental transarterial chemoembolization, showed homogeneous (Group I) or inhomogeneous (Group II) iodized oil accumulation, or whether the tumors were located within the liver segment (Group A) or in a segmental border zone (Group B). Comparison of tumor characteristics between Group IA and the other three groups was performed using the chi-square test. Local recurrence rates were compared among the groups using the Kaplan-Meier estimation and log rank test. RESULTS: Local tumor recurrence occurred in 19 hepatocellular carcinoma tumors (25.7%). There were: 28, 18, 17, and 11 tumors in Group IA, IB, IIA, and IIB, respectively. One of 28 (3.6%) tumors in Group IA, and 18 of 46 (39.1%) tumors in the other three groups showed local recurrence. Comparisons between Group IA and the other three groups showed that the tumor characteristics were similar. One-, two-, and three-year estimated local recurrence rates in Group IA were 0%, 11.1%, and 11.1%, respectively. The difference between Group IA and the other three groups was statistically significant (p = 0.000). CONCLUSION: An acceptably low rate of local recurrence was observed for small or intermediate nodular tumors located within the liver segment with homogeneous iodized oil accumulation. PMID- 17420629 TI - Variable CT findings of epithelial origin ovarian carcinoma according to the degree of histologic differentiation. AB - OBJECTIVE: We wanted to evaluate the CT findings of epithelial origin ovarian carcinoma according to the degree of histologic differentiation. MATERIALS AND METHODS: This study enrolled 124 patients with 31 well differentiated, 44 moderately differentiated and 95 poorly differentiated carcinomas with epithelial origin. The CT images were retrospectively evaluated with regard to bilateral ovarian involvement, the tumor's nature, lymphadenopathy, adjacent organ invasion, peritoneal tumor seeding, a large amount of ascites and distant metastasis. In cystic, predominantly cystic and mixed tumors, the tumor wall, septa, papillary projection and necrosis in the solid portion were assessed. RESULTS: Bilateral ovarian involvement was more common in the poorly (48%) and moderately (42%) differentiated carcinomas than in the well differentiated carcinomas (7%) (p < 0.05). The frequency of a predominantly solid or solid nature was greater in the moderately and poorly differentiated carcinomas than in the well differentiated carcinomas (p < 0.0001). In the 87 tumors with a cystic, predominantly cystic or mixed nature, septa greater than 3 mm, papillary projection and necrosis in the solid portion were more common in the poorly differentiated carcinoma (91%, 91% and 77%, respectively) than in the moderately (64%, 68% and 34%, respectively) and well differentiated carcinomas (63%, 47% and 27%, respectively) (p < 0.05). Lymphadenopathy, organ invasion, tumor seeding and a large amount of ascites were more common in the poorly differentiated carcinomas (38%, 27%, 73% and 69%, respectively) than in the moderately (13%, 10%, 48% and 45%, respectively) and well differentiated carcinomas (3%, 0%, 10% and 17%, respectively) (p < 0.05). CONCLUSION: Epithelial origin ovarian carcinoma shows different CT findings according to the degree of histologic differentiation. PMID- 17420630 TI - Effects of intraluminal irradiation with Holmium-166 for TIPS stenosis: experimental study in a swine model. AB - OBJECTIVE: We wanted to evaluate the effectiveness of intraluminal irradiation with Holmium-166 ((166)Ho) for reducing the pseudointimal hyperplasia (PIH) in the transjugular intrahepatic portosystemic shunt (TIPS) tract in a swine model. MATERIALS AND METHODS: TIPS was performed in 12 domestic pigs, after the creation of portal hypertension by intraportal injection of a mixture of N-butyl-2 cyanoacrylate (NBCA) and lipiodol. Five pigs first underwent intraluminal irradiation (30 Gy) in the parenchymal tract with using a (166)Ho solution-filled balloon catheter, and this was followed by the placement of a nitinol stent in the TIPS tract. For the seven control pigs, the balloon was filled with saline and contrast media mixture. Two weeks later, follow-up portography and histological analysis were performed. RESULTS: TIPS was successfully performed in all twelve pigs with achieving artificially induced portal hypertension. Portography performed two weeks after TIPS showed the patent tracts in the TIPS tracts that were irradiated with (166)Ho (5/5, 100%), whereas either completely (5/6, 83.3%) or partially (1/6, 16.7%) occluded TIPS were seen in the seven pigs of the nonirradiated control group, except in one pig that experienced periprocedural death due to bleeding. Histological analysis showed a statistically significant difference for the maximal PIH (irradiated: 32.8%, nonirradiated: 76.0%, p < 0.001) between the two groups. CONCLUSION: Intraluminal irradiation with 30 Gy of (166)Ho for TIPS significantly improved the TIPS patency in a swine model of portal hypertension during a 2-week period of follow up. PMID- 17420631 TI - Transcatheter arterial embolization of renal VX-2 carcinoma: ethiodol-ethanol capillary embolization combined with carboplatin. AB - OBJECTIVE: We wanted to determine whether transcatheter Ethiodol-based capillary embolization in combination with carboplatin could improve the efficiency of a 1:1 Ethiodol-ethanol mixture (EEM) to ablate kidneys that been inoculated with VX 2 carcinoma. MATERIALS AND METHODS: The right kidney in 34 New Zealand white rabbits were inoculated with fresh VX-2 tumor fragments. One week later, the kidneys were subjected to transarterial treatment (4-5 rabbits/group): Saline infusion (Group 1); carboplatin infusion (5 or 10 mg, Groups 2A and 2B); carboplatin-Ethiodol (CE) alone (Group 3) and followed by main renal artery occlusion with ethanol (RAO) (Group 4); carboplatin-EEM (C-EEM) followed by RAO (Group 5); carboplatin infusion followed by EEM plus RAO (Group 6); and EEM followed by RAO (Group 7). The animals were followed for up to 3-weeks. The treated kidneys were evaluated angiographically and macroscopically. The kidneys that showed successful embolization macroscopically were entirely cut into serial sections, and these were examined microscopically. Histologically, the kidneys were evaluated on the basis of the residual tumor found in the serial sections. RESULTS: The results obtained with carboplatin infusion alone (Groups 2A and 2B) and CE without RAO (Group 3) were similar to those of the control animals (Group 1). Kidneys from Groups 4-7 demonstrated macroscopically successful embolization with histologically proven complete renal parenchyma infarction; however, some residual tumor was evident in all but one animal. CONCLUSION: None of the Ethiodol-based modalities combined with locoregional carboplatin were more efficacious for tumor ablation than EEM alone. PMID- 17420632 TI - Morton neuroma: evaluated with ultrasonography and MR imaging. AB - OBJECTIVE: The purpose of this study was to compare the diagnostic accuracy of both ultrasonography (US) and magnetic resonance imaging (MRI) for the assessment of Morton neuroma. MATERIALS AND METHODS: Our study group was comprised of 20 neuromas from 17 patients, and the neuromas were confirmed on surgery following evaluation with US, MRI, or both US and MRI. The diagnostic criterion for Morton neuroma, as examined by US, was the presence of a round or ovoid, well-defined, hypoechoic mass. The diagnostic criterion, based on MR imaging, was a well defined mass with intermediate to low signal intensity on both the T1- and T2 weighted images. The retrospective comparison between the sonographic and MR images was done by two experienced radiologists working in consensus with the surgical and pathologic correlations. RESULTS: The detection rate of Morton neuroma was 79% for 14 neuromas from 11 patients who had undergone US followed by an operation. The detection rate was 76% for 17 neuromas from 15 patients who had undergone MRI and a subsequent operation. The mean size of the examined neuromas was 4.9 mm on the US images and it was 5.1 mm on the MRI studies. Ten neuromas (71%) were 5 mm or less as measured by US, and three neuromas were not detected, whereas on the MRI analysis, 10 neuromas (59%) were 5 mm or less and four neuromas were not visualized. Among the patients examined during postoperative follow-up, symptoms were completely relieved in 85% and the symptoms were partially relieved in 15%. CONCLUSION: US and MR imaging are comparable modalities with high detection rate for the evaluation of Morton neuroma. PMID- 17420633 TI - The use of magnetic resonance imaging to predict the clinical outcome of non surgical treatment for lumbar intervertebral disc herniation. AB - OBJECTIVE: We wanted to investigate the relationship between the magnetic resonance (MR) findings and the clinical outcome after treatment with non surgical transforaminal epidural steroid injections (ESI) for lumbar herniated intervertebral disc (HIVD) patients. MATERIALS AND METHODS: Transforaminal ESI were performed in 91 patients (50 males and 41 females, age range: 13-78 yrs) because of lumbosacral HIVD from March 2001 to August 2002. Sixty eight patients whose MRIs and clinical follow-ups were available were included in this study. The medical charts were retrospectively reviewed and the patients were divided into two groups; the successful (responders, n = 41) and unsatisfactory (non responders, n = 27) outcome groups. A successful outcome required a patient satisfaction score greater than two and a pain reduction score greater than 50%. The MR findings were retrospectively analyzed and compared between the two groups with regard to the type (protrusion, extrusion or sequestration), hydration (the T2 signal intensity), location (central, right/left central, subarticular, foraminal or extraforaminal), and size (volume) of the HIVD, the grade of nerve root compression (grade 1 abutment, 2 displacement and 3 entrapment), and an association with spinal stenosis. RESULTS: There was no significant difference between the responders and non-responders in terms of the type, hydration and size of the HIVD, or an association with spinal stenosis (p > 0.05). However, the location of the HIVD and the grade of nerve root compression were different between the two groups (p < 0.05). CONCLUSION: MRI could play an important role in predicting the clinical outcome of non-surgical transforaminal ESI treatment for patients with lumbar HIVD. PMID- 17420634 TI - Transarterial embolization with Berenstein Liquid Coils and N-butyl cyanoacrylate in a vein of Galen aneurysmal malformation: a case report. AB - A 12-week-old baby with a vein of Galen aneurysmal malformation (VGAM) was successfully treated with performing transarterial microcatheter-directed embolization with Berenstein Liquid Coils and n-butyl cyanoacrylate in the feeding arteries. Post-procedure angiography showed a marked decrease of the blood flow into the dilated vein of Galen. Three months later, follow-up angiography showed that the vein of Galen aneurysmal malformation had totally disappeared, and the baby recovered very well without any sequelae. We report here on this interesting case along with a review of the relevant literature, and we aim to enhance physicians' awareness of the treatment for VGAMs. PMID- 17420635 TI - Actinomycosis of the gallbladder mimicking carcinoma: a case report with US and CT findings. AB - We describe a case of actinomycosis of the gallbladder mimicking carcinoma. Sonography showed a hypoechoic mass replacing gallbladder lumen and engulfing a stone; contrast-enhanced computed tomography showed a heterogeneously enhanced thickened gallbladder wall with subtle, disrupted luminal surface enhancement, which formed a mass. As a result of the clinical and radiologic presentation, our impression was of gallbladder carcinoma. Actinomycosis should be included in the differential diagnosis when sonography and computed tomography findings show a mass engulfing the stone in the gallbladder and extensive pericholecystic infiltration with extension to neighboring abdominal wall muscle. PMID- 17420636 TI - Transarterial embolization of an inferior genicular artery pseudoaneurysm with arteriovenous fistula after arthroscopy. AB - Arthroscopic meniscectomy of the knee is generally a safe and effective procedure with a low rate of vascular complications. We report here on a unique case of a 55-year-old man with a lateral inferior genicular artery pseudoaneurysm and a concomitant arteriovenous fistula that developed after arthroscopic meniscectomy; this was successfully treated with selective angiographic embolization. This case illustrates the effectiveness of an endovascular approach as a minimally invasive treatment for this uncommon complication that occurs after an arthroscopic procedure. PMID- 17420637 TI - Selective uterine artery embolization for management of interstitial ectopic pregnancy. AB - Interstitial ectopic pregnancy is a rare condition of pregnancy and may be very dangerous if not identified and treated urgently. We report a case of successful treatment of an interstitial pregnancy using selective uterine artery embolization. A 27-year-old woman with interstitial pregnancy was treated by uterine artery embolization after failure of systemic methotrexate treatment. Her serum beta-human chorionic gonadotropin (beta-hCG) was undetectable one month after the therapeutic embolization and transvaginal sonography 31 days after embolization showed normal endometrium and cornu. The patient achieved a normal pregnancy eight months after embolization. PMID- 17420638 TI - Solitary small hepatic angiosarcoma: initial and follow-up imaging findings. AB - We report an uncommon case of solitary, small hepatic angiosarcoma that was initially considered as a hemangioma. We present the imaging findings, with an emphasis on the initial and follow-up CT and MR findings, as well as report on the more suggestive findings of angiosarcoma than those of a hemangioma. PMID- 17420639 TI - Notice of redundant publication. PMID- 17420640 TI - How do I get more involved in STN? PMID- 17420641 TI - Advocacy: the tool of a hero. PMID- 17420644 TI - Ask the expert. The "halo effect" that occurs when trauma centers are in place. PMID- 17420645 TI - Penetrating neck injuries in children. AB - Penetrating neck injuries are uncommon in children, and when they do occur, they can be a challenge to manage. Injuries can be cardiovascular, aerodigestive, and neurovascular. In the past, injuries were explored surgically to evaluate the extent of the injury; studies are now showing that observation and less invasive studies have a positive outcome for the patient. Immediate surgical intervention remains mandatory for the clinically unstable patient, whereas stable patients have studies done that pertain to their signs and symptoms, thus having a conservative nonoperative approach to care. PMID- 17420647 TI - A patient education tool for nonoperative management of blunt abdominal trauma. AB - Blunt trauma is the primary mechanism of injury seen at Charleston Area Medical Center, a rural level I trauma center. Blunt abdominal trauma occurs as a result of various mechanisms. It can be safely managed nonoperatively and is considered to be the standard of care in hemodynamically stable patients. Appropriate patient education before discharge will enable patients to identify complications early and seek appropriate medical care. PMID- 17420648 TI - Continued professional competence and portfolios. AB - It is traditionally assumed that licensure of healthcare professionals means that they are minimally competent. Many nursing specialty organizations offer examinations and other processes for certification, suggesting that certification is associated with continued competency. Can standardized examination for certification and continuing education for recertification ensure continued competency? Continuing education and testing provide a limited picture of an individual's knowledge and/or skill acquisition in a limited area at one point in time. However, portfolios promote critical thinking, self-assessment, and individual accountability. A portfolio is a portable mechanism for evaluating competencies that may otherwise be difficult to assess. This article summarizes some of the literature addressing portfolios, including aspects of portfolio development process, the value of portfolios versus continuing education for competency assessment, evidence associated with portfolio usage, and suggestions for organizing nursing portfolios. PMID- 17420649 TI - Hey! I'm strapped to this board and can't get up! PMID- 17420650 TI - Body piercing/tattooing and trauma diagnostic imaging: medical myths vs realities. PMID- 17420651 TI - Lessons learned when science meets legislation. PMID- 17420652 TI - "The calm after the storm". AB - The vision of Injury Free Coalition for Kids is that children and families will live in safe communities; our mission is to create them. After the devastation caused by Hurricane Katrina, members of Injury Free Coalition for Kids went to Biloxi, Miss, to develop a coalition of community stakeholders in order to build playgrounds for 2 neighborhoods in Biloxi. As child advocates, we know that play is the work of children, and through play, children exposed to disaster can rediscover joy. PMID- 17420653 TI - Donation after cardiac death: lessons learned. AB - Despite the increasingly positive outcome of organ transplantation as an accepted treatment of end-stage organ diseases, an average of 15 people die each day awaiting organ transplantation. According to the United Network for Organ Sharing, there are more than 90,000 people in the United States waiting for an organ transplant. In the United States, less than 1% of all deaths are attributed to brain death. A single brain-dead organ donor has the potential to save up to 8 individuals by donating organs and providing up to 50 people with tissue and cornea transplants. The reality is that the source of available brain-dead donors does not meet the needs of the growing waiting list. To help deal with the increasing demand for organs, donation after cardiac death has been reintroduced to families of patients with catastrophic brain injuries. Families have the right to be informed of all potential end-of-life options, including that of organ donation and the use of donation after cardiac death when appropriate. Hospitals and healthcare workers must be committed to provide the option of donation after cardiac death for both donor families and transplant recipients. The purpose of this article is to examine the process of implementing a donation after cardiac death policy in a 1,061-bed tertiary care hospital with level I trauma designation. PMID- 17420654 TI - Have we seen the last of sharp retractor use in surgical, emergent, or trauma procedures? PMID- 17420659 TI - Toll-like receptors: emerging concepts in kidney disease. AB - PURPOSE OF REVIEW: To summarize the recent advances in the role of Toll-like receptors (TLRs) in innate immunity, with a special focus on recent studies addressing the expression and function of TLRs in kidney disease. RECENT FINDINGS: Pathogen-recognition receptors including TLRs mediate immune activation upon pathogen recognition in different extracellular and intracellular compartments. In contrast to professional antigen-presenting cells, renal cells express a limited pattern of TLR (i.e. express TLR1-TLR6 but lack expression of the endosomal TLR7-TLR9). TLRs on renal cells contribute to the innate immune response in renal infection. Furthermore, recent studies provide experimental evidence for the functional role of TLRs in immune complex disease and autoimmunity. Furthermore, the recognition of endogenous molecules released from injured cells such as biglycan or heat-shock proteins may contribute to acute tubular injury and seem to provide adjuvant activity for renal inflammation. Furthermore, TLR7 and TLR9 are involved in the pathogenesis of lupus nephritis. SUMMARY: The field of TLR research elucidates the molecular mechanisms of infection-associated kidney diseases but may also further support the concept that innate immunity significantly contributes to the so-called types of nonimmune kidney diseases. PMID- 17420660 TI - New insights into structural patterns encountered in glomerulosclerosis. AB - PURPOSE OF REVIEW: The term 'focal segmental glomerulosclerosis' covers a variety of diseases with different histopathological patterns. There is a need for clues to interpret histological findings in terms of etiology. Studies in transgenic animal models published in recent years have targeted the podocyte with respect to its impact on the development of glomerulosclerosis. Our aim was to survey those models in an attempt to discover correlations between histopathological patterns and pathogenic mechanisms. RECENT FINDINGS: The most obvious conclusion to draw from recent studies is that virtually all forms of glomerulosclerosis start with a lesion or dysfunction of podocytes. In hereditary glomerular diseases and transgenic animal models, two patterns of glomerular degeneration may be distinguished. All diseases with late onset appear to follow the 'classic' pathway to focal segmental glomerulosclerosis, starting with an adhesion of the tuft to the Bowman's capsule and eventually leading to nephron degeneration. In contrast, those with early onset frequently exhibit changes that indicate a severe dysregulation of podocyte function resulting in diffuse global endocapillary damage (i.e. mesangial expansion and rarefaction of capillaries). SUMMARY: Such insights derived from animal models might be useful in elucidating the mechanisms of multifactorial human diseases like diabetic glomerulopathy. PMID- 17420661 TI - Collapsing glomerulopathy: an inflammatory podocytopathy? AB - PURPOSE OF REVIEW: Collapsing glomerulopathy is a relatively new and debated podocytopathy. Among several conjectures, inflammatory injury orchestrated by podocytes is emerging to explain the pathogenesis of collapsing glomerulopathy. Here, we briefly summarize recent studies in support of this novel and intriguing hypothesis. RECENT FINDINGS: Immunohistochemical analyses of markers conventionally used to demarcate podocytes apart from parietal epithelium identified the parietal podocyte. MafB-deficient mice exhibited abnormal podocyte and macrophage differentiation, suggesting ancestral and functional overlap. These apparent developmental anomalies were detected in studies showing an admixture of hyperplastic podocytes with macrophage epitopes and hyperplastic parietal epithelium in pseudocrescents and in true crescents. Experimental antibody-mediated injury of podocytes could trigger capillary collapse and pseudocrescent formation marked by recruitment of epithelial cells from Bowman's capsule. In contrast, experimental stabilization of hypoxia-inducible factors within podocytes--a known inflammatory response by macrophages--could trigger podocyte proliferation and the formation of true necrotizing crescents. SUMMARY: Preliminary evidence suggests that visceral and parietal podocytes may become macrophage-like inflammatory mediators of proliferative epithelial injury within the glomerulus. This may manifest as collapsing glomerulopathy or crescentic glomerulonephritis--lesions that appear to be anatomically and pathogenically linked. PMID- 17420662 TI - New insights into systemic amyloidosis: the importance of diagnosis of specific type. AB - PURPOSE OF REVIEW: This review aims to summarize recent developments in the area of systemic amyloidoses with emphasis on pathologic diagnosis. RECENT FINDINGS: In recent years, management of amyloidosis has shifted from a purely supportive approach to quite diverse, radical and aggressive treatments. The central issue is the understanding that treatment of systemic amyloidoses depends on the molecular type of the amyloid protein. In the United States and the Western world, AL-amyloidosis is the most prevalent type of systemic amyloidosis, but hereditary amyloidoses are being diagnosed with increasing frequency; genetics also plays a role in a subset of familial AA amyloidoses. The biggest challenge is in the diagnosis of AL-type with confidence and in differentiation of AL and hereditary amyloidoses. While careful clinico-pathologic correlation is recommended for all patients with amyloidosis, it is, in itself, not a substitute for amyloid typing. SUMMARY: The diagnosis of the amyloid type ultimately depends on the examination of the amyloid protein within the deposits. The role of immunohistochemistry - the current standard of care in amyloid typing - is evolving with emergence of alternative biochemical methods. Amyloid, being essentially a protein disorder, presents an attractive venue for the application of proteomics methodologies, despite their inherent complexities. PMID- 17420663 TI - Dense deposit disease: new insights. AB - PURPOSE OF REVIEW: Dense deposit disease is a rare but devastating disease primarily affecting children. This review focuses on new information regarding the pathophysiology of dense deposit disease, its appearance histopathologically, its relationship to other diseases including macular degeneration and acquired partial lipodystrophy and potential new therapies. RECENT FINDINGS: The microscopic features of dense deposit disease have been separated into five patterns with only about 25% of patients showing membranoproliferative features. The subtle interplay between genetic changes in complement regulatory proteins and dysregulation of the alternative pathway of complement is now more evident. Haplotype mapping has shown at-risk phenotypes of complement factor H associated with the development of dense deposit disease. Treatment protocols are empiric and not very effective. New information on complement inhibitors and plasma exchange, however, has brought hope for new therapies in the near future. SUMMARY: Understanding of the pathology and the pathophysiology of dense deposit disease has advanced rapidly in the last decade. New efforts in genetic mapping along with the development of novel inhibitors of the complement system will lead to improved care for patients afflicted with this uncommon condition. PMID- 17420664 TI - Preeclampsia: new insights. AB - PURPOSE OF REVIEW: Preeclampsia is a disorder of gestation characterized by hypertension and proteinuria and can be complicated by eclamptic seizures. This review describes recent advances in the role of the renin-angiotensin system and angiogenic and anti-angiogenic factors of placental origin in its pathogenesis. RECENT FINDINGS: Deficient uteroplacental perfusion has been recognized to be a feature in all preeclampsia syndromes. Increased renin expression observed in humans and animal models supports the concept that activation of the decidual renin-angiotensin system may mediate the pathogenesis of preeclampsia. Novel angiotensin II-related biomolecular mechanisms, angiotensin II type 1-B2 receptor heterodimerization and autoantibody against angiotensin II type 1 have recently been described in preeclampsia. New evidence suggests that vascular endothelial growth factor and its receptors, antagonists, and reduced placental growth factor may play a role in the development of proteinuria and other renal injury-mediated manifestations in preeclampsia. SUMMARY: Vascular maladaptation, with increased vasomotor tone, endothelial dysfunction, increased sensitivity to angiotensin II and norepinephrine, and multiorgan dysfunction seen in preeclampsia, may be explained by angiotensin II-mediated mechanisms. Future investigations need to define the mechanism of activation of the decidual renin-angiotensin system and the release of placental factors in the pathogenesis of preeclampsia. PMID- 17420665 TI - Linking the population epidemiology of acute renal failure, chronic kidney disease and end-stage renal disease. PMID- 17420667 TI - Obesity and kidney disease: a big dilemma. AB - PURPOSE OF REVIEW: Obesity is the number one preventable risk factor for chronic kidney disease. Obesity is, however, associated with improved survival in patients with end-stage renal disease (ESRD). RECENT FINDINGS: Multiple observational studies have documented an association between obesity and risk of kidney disease even after adjustment for obesity-related co-morbid conditions, including diabetes. Prevalence of a body mass index of at least 35 kg/m among incident dialysis patients has increased by 64% over the past decade, and if trends continue 20% of all patients will initiate dialysis with this degree of obesity. Weight loss improves glomerular hemodynamics in morbidly obese adults and may retard progression of chronic kidney disease. In contrast, once a patient reaches ESRD, the degree of adiposity correlates with survival, and weight loss may not necessarily be beneficial. SUMMARY: Weight loss appears to be beneficial in obese patients both with and without chronic kidney disease. The safety of intentional weight loss in obese ESRD patients, however, remains questionable. The preclusion of obese ESRD patients from kidney transplantation needs to be readdressed and more studies are needed to determine effective strategies for addressing the obesity epidemic in the chronic kidney disease and ESRD populations. PMID- 17420666 TI - The incidence and prognostic significance of acute kidney injury. AB - PURPOSE OF REVIEW: Acute kidney injury is an increasingly common and potentially catastrophic complication in hospitalized patients. This review summarizes the major epidemiologic studies that have informed our understanding of the incidence and prognostic significance of acute kidney injury. RECENT FINDINGS: Early observational studies from the 1980s and 1990s established the general epidemiologic features of acute kidney injury, including the incidence, prognostic significance and predisposing medical and surgical conditions. Recent multicenter observational cohorts and administrative databases have enhanced our understanding of the overall disease burden of acute kidney injury and trends in its epidemiology. An increasing number of clinical studies focusing on specific types of acute kidney injury (e.g. following exposure to intravenous contrast, sepsis and major surgery) have provided further details into this heterogeneous syndrome. SUMMARY: In light of the increasing incidence and prognostic significance of acute kidney injury, new strategies for prevention and treatment are desperately needed. PMID- 17420668 TI - Understanding estimated glomerular filtration rate: implications for identifying chronic kidney disease. AB - PURPOSE OF REVIEW: Glomerular filtration rate (GFR) can be estimated using serum markers such as serum creatinine (SCr) or cystatin C. This review presents new insights into estimated GFR based on theory, validation studies, SCr assay standardization, cystatin C, and longitudinal comparison with measured GFR. RECENT FINDINGS: The estimation of GFR by SCr differs in health and in chronic kidney disease (CKD) due to differences in GFR range and in creatinine production between these two populations. Among populations with normal baseline GFR, there is a more rapid decline in measured GFR than in SCr-based estimated GFR. While elevated SCr is specific for CKD, other disease processes may lead to elevated cystatin C. Validation is improved by refitting equation coefficients to compare populations, recognizing the asymmetry between estimated GFR and measured GFR, and using residual plots instead of Bland-Altman plots to assess bias. SUMMARY: As a screening test, SCr should be interpreted as a marker of CKD probability in the context of the patient's clinical presentation. Measured GFR or creatinine clearance may be helpful in high-risk patients with normal SCr levels. GFR estimating equations should be reserved for patients with identified CKD. Standardized SCr and cystatin C assays are needed. PMID- 17420669 TI - Treatment options for prehypertension. AB - PURPOSE OF REVIEW: Prehypertension is a recognized stage of blood pressure but treatment standards have not yet been established. This review will focus on the treatment considerations in prehypertension. RECENT FINDINGS: Previous approaches to prehypertensive blood pressure have focused on the role of nonpharmacologic modalities alone. The recently published Trial of Preventing Hypertension (TROPHY) is the first study of pharmacologic intervention in this category of blood pressure. SUMMARY: TROPHY forged new territory in the area of pharmacologic treatment for prehypertension. The safety and tolerability of the angiotensin receptor blocker treatment in the trial offer a new option to clinicians attempting to avoid the ravages of hypertension in the patients they treat. Furthermore the stage is properly set for additional research in treatments for prehypertension. PMID- 17420670 TI - Antihypertensive medications and risk of diabetes mellitus. AB - PURPOSE OF REVIEW: Over the past decade post-hoc analyses of clinical trials and observational studies have tended to show that participants treated with thiazide diuretics are at greater risk for newly diagnosed diabetes mellitus than those treated with other medication classes. We review the results of several recent studies on the impact of thiazide-related hyperglycemia and diabetes mellitus on cardiovascular disease outcomes. We also examine the impact of the glucose sparing effects of angiotensin-converting enzyme inhibitors and angiotensin receptor blockers on preventing cardiovascular disease. RECENT FINDINGS: No consistent or conclusive evidence has been found that hyperglycemia or diabetes mellitus in association with thiazide diuretic use is associated with increased cardiovascular disease outcomes. This benign outcome may be a consequence of the fact that only a segment of such diuretic-associated cases is induced by the usual etiologic mechanisms that are associated with classic 'diabetes mellitus'. Likewise, no evidence has been found that the glucose-lowering effect of angiotensin-converting enzyme inhibitors is associated with decreased cardiovascular disease risk. SUMMARY: We conclude that thiazide diuretics are safe to use, even in hypertensive individuals at risk for incident glucose disorders. The use of angiotensin-converting enzyme inhibitors for protection against glucose disorders and subsequent cardiovascular disease remains to be determined. PMID- 17420671 TI - Analyzing the radial pulse waveform: narrowing the gap between blood pressure and outcomes. AB - PURPOSE OF REVIEW: The use of peripheral pressure wave recording devices to estimate central vessel pressure profiles has grown over the past decade. This review will briefly recount the history behind this technology and discuss where it fits into the measurement of vascular properties that may predict outcomes such as death, heart attack, stroke or kidney disease progression. RECENT FINDINGS: Recent cross-sectional studies involving both children with chronic kidney disease and adult patients with cardiovascular disease are reviewed. Importantly there are now at least three prospective series that indicate pulse wave analysis technology may be useful both for endpoint prediction (complementing blood pressure and other standard vascular risk factors) and for informing mechanisms of benefit when clinical trials with similar blood pressure control have discordant outcomes depending on the drug assignment. SUMMARY: Pulse wave analysis is an interesting technology with the plausible benefit of estimating central vascular pressures. Since the principal target organs we seek to protect with our treatments are all connected to the aorta, it may be that a better understanding of target organ risk from an aortic as opposed to a brachial standpoint will help move forward our interventions to reduce vascular disease. PMID- 17420672 TI - The treatment of anemia in chronic kidney disease: understandings in 2006. AB - PURPOSE OF REVIEW: Anemia is a well recognized complication of chronic kidney disease and is associated with significant morbidity. It is important for clinical care to identify appropriate treatments and targets for hemoglobin. This review describes current understandings of the treatment of anemia using the most recent published articles. RECENT FINDINGS: Numerous studies, including observational and randomized control trials, of varying sizes and using both surrogate and hard outcomes have been published. On balance, there is little to support normalization of hemoglobin in the chronic kidney disease population. While some studies have described harm, there are some issues related to overinterpretation based on study trial reporting. The treatment of anemia can be successfully achieved with the use of oral or intravenous iron and erythropoiten stimulating agents. Caution should be exercised when treating those with significant cardiovascular morbidity, and those who require very high doses of erythropoiten-stimulating agents to achieve normal hemoglobin. SUMMARY: Large observational population-based studies continue to demonstrate the association of low hemoglobin with adverse outcomes, and randomized control trials fail to show a benefit of normalized hemoglobin. Anemia therapy does improve quality of life. In the current era of aggressive chronic kidney disease management, it does not appear that anemia therapy attenuates left ventricular growth or changes cardiovascular outcomes. PMID- 17420673 TI - Bibliography. Current world literature. Renal immunology and pathology. PMID- 17420674 TI - Autism in children with 22q11.2 deletion syndrome. PMID- 17420676 TI - False methadone-positive urine drug screens in patients treated with quetiapine. PMID- 17420677 TI - Megaloblastic anemia and osteopenia in an adolescent diagnosed with schizophrenia. PMID- 17420678 TI - Psychiatric disorders in Norwegian 8- to 10-year-olds: an epidemiological survey of prevalence, risk factors, and service use. AB - OBJECTIVE: The Bergen Child Study is a longitudinal study of child mental health from the city of Bergen, Norway. We present methods and results from the first wave of the study, focusing on prevalence of disorders, associations with risk factors, and the use of services. METHOD: The target population included all 9,430 children attending grades 2 to 4 in Bergen schools during the academic year 2002/2003. The main screening instrument was the Strengths and Difficulties Questionnaire, whereas diagnoses were based on the Development and Well-Being Assessment. Information about child and family risk factors and service use was also obtained in this second stage. RESULTS: In the first phase, the teacher Strengths and Difficulties Questionnaire was obtained for 9,155 (97%) of the target children and the matching parent Strengths and Difficulties Questionnaire for 6,297 (67%); 1,011 children (11%) were assessed with the Development and Well Being Assessment in the second phase. The weighted prevalence for any DSM-IV psychiatric disorder was 7.0% (95% confidence interval 5.6%-8.5%). Disorders were associated with age, gender, learning difficulties, family type, and poverty. Although 75% of children with attention-deficit/hyperactivity disorder had been in contact with specialist mental health services, this was true for only 13% of those with pure emotional disorders. CONCLUSIONS: The overall prevalence of psychiatric disorders in children is relatively low in this Norwegian sample, when assessed with the Development and Well-Being Assessment. Children with emotional disorders have limited access to specialist services. PMID- 17420679 TI - Child, family, school and community risk factors for poor mental health in Brazilian schoolchildren. AB - OBJECTIVE: To identify risk factors for poor child mental health in the southeastern Brazilian municipality of Taubate. METHOD: In 2001 we carried out a representative school-based survey of Brazilian schoolchildren ages 7 to 14 years (response rate, 83%). We collected extensive data on child mental health and on potential risk and protective factors at baseline (N = 1,112) and collected follow-up information on mental health outcome after 3 years (N = 760). We used the Strengths and Difficulties Questionnaire as a dimensional measure of mental health at both time points and the Development and Well-Being Assessment to diagnose psychiatric disorder at baseline. RESULTS: Cross-sectionally, both dimensional and diagnostic measures of child psychopathology were associated with dangerous area, nontraditional family, parental stress, harsh physical punishment, poor general health, low IQ, repeating a year at school, and male gender. There were different cross-sectional associations with different types of psychopathology. Psychological difficulties were moderately persistent across time. Younger age, lower maternal education, and lower child's IQ predicted a worse prognosis even after adjusting for initial psychopathology. CONCLUSIONS: The multiple factors associated with Brazilian child mental health are broadly in line with previous findings elsewhere. Public health programs to foster child mental health need multiple targets, but may benefit a wide range of the population. PMID- 17420680 TI - Further evidence of the reliability and validity of DSM-IV ODD and CD in preschool children. AB - OBJECTIVE: To test the reliability and validity of DSM-IV oppositional defiant and conduct disorders (ODD and CD) and symptoms using the Kiddie Disruptive Behavior Disorders Schedule and generate data on the manifestation of symptoms of ODD and CD in 3- to 5-year-old children. METHOD: One hundred twenty-three consecutive referrals to a child and adolescent psychiatry clinic and 100 children recruited from a pediatric clinic and their biological mothers participated. Mothers were interviewed about current ODD and CD symptoms, and mother and teachers completed measures of impairment. Twenty-five percent of the interviews were randomly selected and coded by a second rater to test interrater reliability. A separate sample of 31 children was recruited for a 1-week test retest study. RESULTS: Overall, reliability of ODD and CD symptoms as measured by interrater and test-retest reliability was moderate to high. Reliability was not affected by age or sex of the child. Validity was demonstrated via associations with impairment and differentiation between referred and nonreferred children. CONCLUSIONS: Most DSM-IV ODD and CD symptoms can be reliably and validly assessed in preschool children. Some developmental modifications are recommended. The results add to the literature on best methods of assessment of preschool psychopathology. PMID- 17420681 TI - Family-based cognitive-behavioral therapy for pediatric obsessive-compulsive disorder: comparison of intensive and weekly approaches. AB - OBJECTIVE: To examine the relative efficacy of intensive versus weekly cognitive behavioral therapy (CBT) for children and adolescents with obsessive-compulsive disorder (OCD). METHOD: Forty children and adolescents with OCD (range 7-17 years) were randomized to receive 14 sessions of weekly or intensive (daily psychotherapy sessions) family-based CBT. Assessments were conducted at three time points: pretreatment, posttreatment, and 3-month follow-up. Raters were initially blind to randomization. Primary outcomes included scores on the Children's Yale-Brown Obsessive-Compulsive Scale, remission status, and ratings on the Clinical Global Impression-Severity and Clinical Global Improvement scales. Secondary outcomes included the Child Obsessive Compulsive Impact Scale Parent Rated, Children's Depression Inventory, Multidimensional Anxiety Scale for Children, and Family Accommodation Scale. Adjunctive pharmacotherapy was not an exclusion criterion. RESULTS: Intensive CBT was as effective as weekly treatment with some advantages present immediately after treatment. No group differences were found at follow-up, with gains being largely maintained over time. Although no group x time interaction was found for the Children's Yale-Brown Obsessive Compulsive Scale (F(1,38) = 2.2, p = .15), the intensive group was rated on the Clinical Global Impression-Severity as less ill relative to the weekly group (F(1,38) = 9.4, p < .005). At posttreatment, 75% (15/20) of youths in the intensive group and 50% (10/20) in the weekly group met remission status criteria. Ninety percent (18/20) of youths in the intensive group and 65% (13/20) in the weekly group were considered treatment responders on the Clinical Global Improvement (chi1(2) = 3.6, p = .06). CONCLUSIONS: Both intensive and weekly CBT are efficacious treatments for pediatric OCD. Intensive treatment may have slight immediate advantages over weekly CBT, although both modalities have similar outcomes at 3-month follow-up. PMID- 17420682 TI - Venlafaxine ER for the treatment of pediatric subjects with depression: results of two placebo-controlled trials. AB - OBJECTIVE: The safety, efficacy, and tolerability of venlafaxine extended release (ER) in subjects ages 7 to 17 years with major depressive disorder were evaluated in two multicenter, randomized, double-blind, placebo-controlled trials conducted between October 1997 and August 2001. METHOD: Participants received venlafaxine ER (flexible dose, based on body weight; intent to treat, n = 169) or placebo (intent to treat, n = 165) for up to 8 weeks. The primary efficacy variable was the change from baseline in the Children's Depression Rating Scale-Revised score at week 8. RESULTS: There were no statistically significant differences between venlafaxine ER and placebo on the Children's Depression Rating Scale-Revised in either study. A post hoc age subgroup analysis of the pooled data showed greater improvement on the Children's Depression Rating Scale-Revised with venlafaxine ER than with placebo (-24.4 versus -19.9; p = .022) among adolescents (ages 12-17), but not among children (ages 7-11). The most common adverse events were anorexia and abdominal pain. Hostility and suicide-related events were more common in venlafaxine ER-treated participants than in placebo-treated participants. There were no completed suicides. CONCLUSIONS: Venlafaxine ER may be effective in depressed adolescents. However, its safety and efficacy in pediatric patients has not been established. Prescribers should monitor for signs of suicidal ideation and hostility in pediatric patients taking venlafaxine ER. PMID- 17420683 TI - Data safety monitoring boards and other study methodologies that address subject safety in "high-risk" therapeutic trials in youths. PMID- 17420684 TI - Phenomenology and correlates of complicated grief in children and adolescents. AB - OBJECTIVE: To describe the phenomenology of complicated grief (CG) in parentally bereaved children and adolescents and to examine its correlates. METHOD: This is a preliminary report from an ongoing 5-year, population-based, longitudinal study of the impact of parental loss on family members. Analyses of cross-sectional data at intake are presented. The sample consists of 129 children and adolescents of parents who died by suicide, accident, or sudden natural death. Their average age is 13.3 +/- 3.1 years (range 7-18 years). A modified version of the Inventory of Complicated Grief-Revised (ICG-R) was administered and its factor structure, internal consistency, and convergent and discriminant validity were examined. RESULTS: CG was significantly related to functional impairment even after controlling for current depression, anxiety, and posttraumatic stress disorder. CG was also associated with other measures of psychopathology, including suicidal ideation. CONCLUSIONS: In this preliminary analysis, CG appears to be a clinically significant syndrome in children and adolescents. Longitudinal data will help to clarify the prognostic significance of CG as well as to examine the interrelationship of CG and other psychopathology over time. PMID- 17420685 TI - Case series: mental health needs and perspectives of rural children reared by parents who abuse methamphetamine. AB - OBJECTIVE: This case-based, mixed-methods study was undertaken to understand the perspectives and mental health needs of rural children exposed to parental methamphetamine abuse. METHOD: Participants were 23 children involved with a state child protective agency because of parental methamphetamine abuse. A semistructured interview provided information on children's perspectives of their families. Information on children's mental health needs was obtained from the Child Behavior Checklist and Trauma Symptom Checklist. Case records and caseworker reports provided information on children's family experiences. RESULTS: Children described emotional pain; few social resources for coping with emotions, problem solving, or talking about their experiences; and avoidant or passive coping skills. Sixty-five percent of children evidenced significant dissociative or posttraumatic symptoms on standardized assessments; 57% had other significant emotional and behavioral problems. Challenges to understanding children's perspectives included children's perceptions that talking about methamphetamine abuse was taboo and underreporting of significant symptoms on the Trauma Symptom Checklist. CONCLUSIONS: The high rate of mental health problems suggests the need for nontraditional strategies for services delivery in rural areas that are targeted toward these vulnerable children. Early identification and treatment of mental health problems should be a priority. Clinicians should be alert to the complexities in assessing children's mental health needs. PMID- 17420686 TI - Pubertal stage and deliberate self-harm in adolescents. AB - OBJECTIVE: To ascertain the association between pubertal stage and deliberate self-harm. METHOD: Cross-sectional survey of 12- to 15-year-olds in 300 secondary schools in the U.S. state of Washington in February-April 2002 and the Australian state of Victoria in June-August 2002. A total of 3,332 students in grades 7 and 9 provided complete data on episodes of deliberate self-harm in the previous 12 months and pubertal stage. Pubertal stage was assessed with the Pubertal Development Scale. RESULTS: The prevalence of deliberate self-harm was 3.7% with a more than twofold higher rate in females. Late puberty was associated with a more than fourfold higher rate of self-harm (odds ratio 4.6, 95% confidence interval 1.5-14) after adjustment for age and school grade level. In contrast age had a protective association (odds ratio 0.7, confidence interval 0.4-1.0). The sharpest rises in prevalence across puberty were for self-laceration and self poisoning in females. Higher rates of depressive symptoms, frequent alcohol use, and initiation of sexual activity largely accounted for the association between self-harm and pubertal stage in multivariate models. CONCLUSIONS: Puberty is associated with changes in the form and frequency of self-harm. For adolescents with a gap between puberty and brain development, risk factors such as early sexual activity and substance abuse may be particularly potent. PMID- 17420687 TI - MRI neuroanatomy in young girls with autism: a preliminary study. AB - OBJECTIVE: To test the hypothesis that young girls and boys with autism exhibit different profiles of neuroanatomical abnormality relative to each other and relative to typically developing children. METHOD: Structural magnetic resonance imaging was used to measure gray and white matter volumes (whole cerebrum, cerebral lobes, and cerebellum) and total brain volume in nine girls (ages 2.29 5.16) and 27 boys (ages 1.96-5.33) with autism and 14 girls (ages 2.17-5.71) and 13 boys (ages 1.72-5.50) with typical development. Structure size and the relationship between size and age were examined. Diagnostic and cognitive outcome data were obtained after the children reached 4 to 5 years of age. RESULTS: Girls with autism exhibited nearly every size-related abnormality exhibited by boys with autism. Furthermore, additional sites of abnormality were observed in girls, including enlargement in temporal white and gray matter volumes and reduction in cerebellar gray matter volume. Significant correlations were observed between age and white matter volumes (e.g., cerebral white matter rs = 0.950) for the girls with autism, whereas no significant age-structure size relationships were observed for the boys with autism. CONCLUSIONS: Results suggest sex differences in etiological factors and the biological time course of the disorder. PMID- 17420688 TI - Glutamine and glutamate levels in children and adolescents with bipolar disorder: a 4.0-T proton magnetic resonance spectroscopy study of the anterior cingulate cortex. AB - OBJECTIVE: The purpose of this study was to use proton magnetic resonance spectroscopy, at 4.0 T, to explore the glutamine and glutamate levels in the anterior cingulate cortex of children and adolescents with bipolar disorder (BPD; medicated and unmedicated) and healthy comparison subjects (HCSs). We hypothesized that unmedicated children with BPD would have reduced glutamine and glutamate levels compared with HCSs and medicated children with BPD. METHOD: Spectra were acquired from the anterior cingulate cortex in 22 children and adolescents with DSM-IV-TR BPD, type 1 (13 female: age 12.6 +/- 4.4 years: 7 of the subjects with BPD were unmedicated at the time of the scan) and 10 HCSs (7 female: age 12.3 +/- 2.5 years). RESULTS: Unmedicated subjects with BPD had significantly lower glutamine levels than HCSs or medicated subjects with BPD. There were no differences in glutamate levels between the three groups. CONCLUSIONS: These results are consistent with there being an abnormality in anterior cingulate cortex glia in untreated children and adolescents with BPD. The results of this pilot study may be important in helping us better understand the pathophysiology of child and adolescent BPD. In addition, this observation may help to develop better and more targeted treatments, in particular those affecting the metabolism of glutamine, perhaps by regulation of glutamine synthetase activity. PMID- 17420689 TI - Similar autonomic responsivity in boys with conduct disorder and their fathers. AB - OBJECTIVE: Antisocial behavior frequently occurs in families. This study investigated whether autonomic hypoarousal and hyporesponsivity, which have been observed in antisocial individuals of all ages, are passed from fathers to male offspring. METHOD: The study included 44 boys with early-onset conduct disorder and 36 healthy controls (8 to 13 years old) together with their biological fathers. Resting heart rate and nonspecific skin conductance fluctuations were assessed as arousal measurements, with electrodermal responses and heart rate changes to pictorial stimuli serving as response measurements. In addition, boys and fathers were subjected to psychometric measurements of antisocial behavior. RESULTS: The fathers of boys with CD showed significantly lower electrodermal responses to pictures of either emotional quality than the fathers of controls (p between.015 and < .001), and they revealed fewer nonspecific skin conductance fluctuations (p = .001). In addition, they reported a more aggressive, hostile, and impulsive behavioral style. Psychophysiological measurements were highly correlated between fathers and sons (p between .007 and < .001), and fathers' autonomic responses accounted for group differences in electrodermal responses between boys (p < .001). CONCLUSIONS: The fathers of boys with conduct disorder exhibited an abnormal psychophysiological response pattern similar to that of their sons. High father-son correlations in psychophysiological measures raise the question of whether autonomic abnormalities may constitute a biological mediator through which the disposition for antisocial behavior is transmitted within families. PMID- 17420690 TI - Tumor necrosis factor antagonists: preliminary evidence for an emerging approach in the treatment of ocular inflammation. AB - The anti-tumor necrosis factor (TNF) monoclonal antibody infliximab and the soluble TNF receptor etanercept inhibit the pleiotropic actions of TNF and are widely used for the treatment of rheumatoid arthritis (RA), juvenile idiopathic arthritis (JIA), spondyloarthropathies (SpA), Crohn's disease, and psoriasis with an acceptable safety profile. A pathogenetic role of TNF in ocular inflammatory conditions has recently emerged from small trials reporting preliminary results on the efficacy of these agents in patients with noninfectious uveitis, regardless of the origin of the disease. The authors review the published experience, derived mostly from investigator-sponsored trials and uncontrolled case series, on the use of TNF antagonists in approximately 280 patients with various ocular conditions who were inadequately controlled on currently available therapy. These reports suggest that TNF antagonists, mainly infliximab, which may have better efficacy than etanercept, are useful in the treatment of ocular inflammation associated with Adamantiades-Behcet's disease, RA, JIA, SpA, Crohn's, sarcoidosis, and Graves' disease ophthalmopathy. Infliximab was also beneficial in small numbers of patients with idiopathic uveitis or scleritis, birdshot retinochoroiditis, uveitic and diabetic cystoid macular edema, and age related macular degeneration. The currently available data are nonrandomized and thus preliminary, providing the foundation and justification for randomized trials to assess efficacy and safety. Until such results are available, knowledge regarding the use of anti-TNF regimens in ophthalmology is incomplete. However, the preliminary evidence points to a growing optimism for targeting TNF in patients with ocular inflammation. PMID- 17420691 TI - Microstructural abnormalities in MEWDS demonstrated by ultrahigh resolution optical coherence tomography. AB - BACKGROUND: Histopathological studies of acute multiple evanescent white dot syndrome (MEWDS) have not been reported because of the transient and benign nature of the disease. Ultrahigh resolution optical coherence tomography (UHR OCT), capable of high resolution in vivo imaging, offers a unique opportunity to visualize retinal microstructure in the disease. METHODS: UHR-OCT images of the maculae of five patients with MEWDS were obtained and analyzed. Diagnosis was based on clinical presentation, examination, visual field testing, and angiography. RESULTS: UHR-OCT revealed disturbances in the photoreceptor inner/outer segment junction (IS/OS) in each of the five patients (six eyes) with MEWDS. In addition, thinning of the outer nuclear layer was seen in the case of recurrent MEWDS, suggesting that repeated episodes of MEWDS may result in photoreceptor atrophy. CONCLUSIONS: Subtle disruptions of the photoreceptor IS/OS are demonstrated in all eyes affected by MEWDS. UHR-OCT may be a useful adjunct to diagnosis and monitoring of MEWDS. PMID- 17420692 TI - Intravitreal bevacizumab (Avastin) in the treatment of macular edema secondary to branch retinal vein occlusion. AB - PURPOSE: To report the authors' experience after intravitreal bevacizumab (Avastin, Genentech) injection in patients with macular edema (ME) secondary to branch retinal vein occlusive disease (BRVO). METHODS: A consecutive retrospective review of patients with ME secondary to BRVO who were treated with intravitreal bevacizumab (1.25 mg/0.05 mL). Patients underwent complete ophthalmic evaluation, which included nonstandardized Snellen visual acuity testing, optical coherence tomography (OCT), and/or angiographic testing at baseline and follow-up visits. RESULTS: There were 27 consecutive patients who received intravitreal bevacizumab injections. The mean length of follow-up was 5.3 months (median 6 months, range 3-8 months). The mean visual acuity improved from 20/200(-) at baseline to 20/100(-) at 1 month and 20/100(+) at 3 months and last follow-up (P < 0.001). The mean central 1 mm macular thickness was 478 microm at baseline and decreased to 310, 336, and 332 microm at 1 month, 3 months, and last follow-up (P < 0.001). Patients received an average of two injections (range one to three). No adverse side effects were observed following injections. CONCLUSION: The observed anatomic (by ophthalmic examination, OCT, and/or fluorescence angiography) and visual acuity improvements and lack of serious adverse side effects after intravitreal bevacizumab injection demonstrates, in principle, the potential of bevacizumab for the treatment of ME in this setting. PMID- 17420693 TI - Rebound macular edema following bevacizumab (Avastin) therapy for retinal venous occlusive disease. AB - BACKGROUND: Bevacizumab, a humanized monoclonal antibody to vascular endothelial growth factor (VEGF), has been given via intravitreal injection as an off-label therapy for both neovascular age-related macular degeneration and for macular edema secondary to retinal vascular disease. The authors describe three patients with macular edema secondary to retinal venous occlusion whose edema initially responded to intravitreal bevacizumab but subsequently recurred in excess of that observed before treatment. METHODS: This is a retrospective case series of three patients with macular edema secondary to retinal vein occlusion treated with intravitreal bevacizumab. RESULTS: In all three patients, the rebound retinal edema observed was more pronounced than that present before treatment. CONCLUSION: These cases suggest a potential limitation of using relatively short acting VEGF antagonists in retinal vascular disease of a chronic nature. Frequent repeated injections may be required to prevent a rebound effect with no clearly defined endpoint. Until the long-term safety of multiple injections of these agents is established, the authors recommend caution in using this treatment strategy. PMID- 17420694 TI - Intravitreal bevacizumab for previously treated choroidal neovascularization from age-related macular degeneration. AB - PURPOSE: To report the optical coherence tomography (OCT) findings and visual results in a series of patients treated with intravitreal bevacizumab for choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD), and to determine if a difference in treatment effect exists between previously treated and treatment naive patients. METHODS: A retrospective review of all patients treated with intravitreal bevacizumab for CNV from AMD with visual acuity greater than or equal to 20/320 between September 2005 and February 2006 was performed. OCT data recorded included central macular thickness and the presence or absence of cystic intraretinal fluid, subretinal fluid, or pigment epithelial detachment at the time of the initial injection, at 1-week, 1 month, and 3-month intervals, as well as at the end of follow-up. Visual acuity measurements were recorded using Early Treatment Diabetic Retinopathy Study charts. Any ocular or systemic adverse events were recorded. Statistical analysis was performed to determine if OCT and visual acuity results were significant and to determine if a difference in outcomes existed between previously treated patients and treatment naive patients. RESULTS: Fifty-four eyes of 51 patients treated with intravitreal bevacizumab for CNV from AMD were identified. A total of 178 injections were performed. Mean number of days of follow-up was 138 with 91% of patients having at least 90 days of follow-up. Seventy percent of patients had undergone previous treatment for CNV. The mean number of intravitreal bevacizumab injections per eye was 3.3. Combined treatment with photodynamic therapy was provided in 20% of cases at the initial intravitreal injection. OCT data for all patients revealed an initial mean thickness of 362 mum, which was decreased at 1 week to 278 microm (P = 0.001), 235 microm at 1 month (P < 0.0001), 238 microm at 3 months (P = 0.0004), and 244 microm for the end of follow-up (P < 0.0001). Cystic retinal edema, subretinal fluid, and pigment epithelial detachment resolved in the majority of cases, but pigment epithelial detachment frequently took longer to resolve. Initial mean visual acuity was 20/125 (logMAR 0.8), and final mean visual acuity was 20/100 (logMAR 0.7) (P = 0.03). There was no difference in OCT or visual acuity outcomes (P = 0.62 and P = 0.28, respectively) between previously treated and treatment naive patients. There was no difference in OCT or visual acuity outcomes (P = 0.67 and P = 0.21, respectively) between patients who received combination therapy and those who received monotherapy with intravitreal bevacizumab. No systemic or ocular adverse events were recorded. CONCLUSION: Intravitreal bevacizumab for CNV from AMD results in a rapid decrease in OCT-measured retinal thickness in a majority of cases. Visual acuity also improved in this series, suggesting a potential corresponding visual benefit. This series suggests that previously treated and treatment naive patients have similar outcomes. PMID- 17420695 TI - Intravitreal bevacizumab (Avastin) treatment of neovascular age-related macular degeneration. AB - PURPOSE: To report the change in visual acuity and central retinal thickness by optical coherence tomography (OCT) after intravitreal injections of bevacizumab for the treatment of neovascular age-related macular degeneration (AMD). METHODS: A retrospective case series in a university-based practice evaluated patients with subfoveal choroidal neovascularization (CNV) due to AMD. Patients received intravitreal injections (1.25 mg) of bevacizumab and were monitored monthly with determination of best-corrected ETDRS visual acuity and OCT for persistence of retinal thickening. Eyes were retreated on an "as needed" basis, defined by presence of intraretinal or subretinal fluid. Patients were monitored every 2 months to 3 months for persistence of angiographic leakage. RESULTS: Seventy-nine eyes of 74 consecutive patients received the initial injection of bevacizumab between August 1, 2005, and January 30, 2006. Sixty-eight eyes (86%) of 64 patients had at least 3 months of follow-up. Mean central retinal thickness +/- SD decreased from 304 +/- 83 microm at baseline to 237 +/- 105 microm at 3 months (P = 0.00002). Mean ETDRS visual acuity gained 4 letters from 20/100 at baseline to 20/80-1 at 3 months (P = 0.040). Twenty eyes (25%) appeared to have a sustained response to a single injection and did not require further injections through 3 months. Two patients had a potentially drug-related adverse event (ischemic stroke and myocardial infarction). No serious injection-related adverse events occurred. CONCLUSIONS: Intravitreal bevacizumab injection affects a rapid decrease in retinal thickness to normal or near-normal levels and improvement in visual acuity in eyes with CNV due to AMD. The sustainability of changes in retinal thickness and visual acuity in response to bevacizumab treatment warrant further investigation and long-term follow-up. PMID- 17420696 TI - Intravitreal bevacizumab for refractory pigment epithelial detachment with occult choroidal neovascularization in age-related macular degeneration. AB - PURPOSE: New medications targeting vascular endothelial growth factor show promise in the treatment of wet macular degeneration. This study describes the clinical response and optical coherence tomography (OCT) findings for patients with refractory pigment epithelial detachment (PED) and occult choroidal neovascular membranes (CNVMs) who were treated with intravitreal bevacizumab. METHODS: A retrospective analysis of data for 10 patients with fibrovascular PEDs, initially treated with intravitreal pegaptanib, thermal laser, or photodynamic therapy with or without triamcinolone acetonide administration, was performed. All patients were refractory to previous treatment. They received monthly injections of bevacizumab and were followed by clinical examination, angiography, and OCT. RESULTS: Nine of 10 patients had stable or improved vision. Angiogram findings showed resolution of leakage from CNVMs. OCT demonstrated resolution of the subretinal or intraretinal fluid but persistence of the PED itself. Vision improvement was correlated with OCT changes. CONCLUSION: Intravitreal bevacizumab may be a viable option in treating patients with fibrovascular PEDs. OCT findings suggest that visual improvement is secondary to resolution of subretinal and intraretinal edema without resolution of the PED. PMID- 17420697 TI - Intravitreal bevacizumab (Avastin) for retinal angiomatous proliferation. AB - OBJECTIVE: To evaluate the short-term visual acuity and anatomic responses after intravitreal bevacizumab (Avastin, Genentech) treatment in patients with retinal angiomatous proliferation (RAP). METHODS: The authors conducted a retrospective review of consecutive patients with newly diagnosed or recurrent RAP treated with intravitreal bevacizumab (1.25 mg) during a 3-month period. Complete ocular examination was performed at baseline and follow-up visits. Interval data were analyzed statistically at 1 and 3 months follow-up. RESULTS: Twenty-three eyes of 23 patients underwent intravitreal bevacizumab treatment. The mean age of patients was 81.1 years, median baseline visual acuity of treated eyes was 20/80 (range 20/25-20/800), and mean baseline central macular thickness was 335 mum (optical coherence tomography was available for 22 eyes). Nine eyes had retinal pigment epithelial detachments (PEDs) at baseline. At 1-month follow-up, the median acuity improved to 20/60 (range 20/30-20/400) (P < 0.001), mean central macular thickness decreased to 202 microm (P < 0.001), and PED was present in only 2 eyes (P = 0.016). Seven of 23 eyes at 1 month (30.4%) had improved visual acuity, defined as halving of the visual angle, and no eyes had worse acuity, defined as doubling of the visual angle. Of the 17 eyes available for 3-month follow-up, 5 eyes (29.4%) had better visual acuity, 1 eye (5.9%) had worse acuity, and the remaining 11 (64.7%) had the same acuity. The median visual acuity at month 3 was 20/60 (range 20/25-20/400). There were no thromboembolic phenomena, endophthalmitis cases, retinal detachments, or any other adverse events. CONCLUSION: Treatment of RAP with intravitreal bevacizumab during this retrospective review resulted in a significant decrease in macular thickness and improvement or stabilization of visual acuity. Further long-term investigation is warranted given the promising short-term results. PMID- 17420698 TI - Photodynamic therapy and high-dose intravitreal triamcinolone to treat exudative age-related macular degeneration: 2-year outcome. AB - PURPOSE: To evaluate the efficacy of photodynamic therapy (PDT) and high-dose intravitreal triamcinolone acetonide (TA) injection to treat choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD) at the 2-year follow-up. METHODS: In this prospective, consecutive, comparative, nonrandomized, interventional case series, 30 eyes of 30 consecutive patients with subfoveal CNV associated with AMD were treated by PDT followed by intravitreal injection of 19.4 +/- 2.1 mg TA. Fifteen eyes were naive to treatment (group 1), and 15 had been treated previously by PDT alone (group 2). A group of 15 eyes of 15 patients treated by PDT alone served as controls. The number of Snellen lines gained or lost and PDT sessions were evaluated. RESULTS: Best-corrected visual acuity (BCVA) did not change significantly in group 1 from baseline (0.0 +/- 3.4 Snellen line; range, -5 to 9 Snellen lines; P = 0.81); group 2 lost an average -0.6 +/- 2.5 line (range, -7 to 3 Snellen lines) (P = 0.41), and the control group lost an average of -2.2 +/- 3.4 lines (range, -8 to 2 Snellen lines) (P = 0.03, Wilcoxon signed rank test). The average number of PDT sessions during the 24-month follow-up was 1.9, 1.2, and 3.9 for group 1, group 2, and the control group, respectively. CONCLUSION: Two years after combined PDT/high-dose intravitreal TA to treat AMD-associated CNV, final BCVA was stable, and the need for retreatment was reduced compared with historical controls. PMID- 17420699 TI - Phacovitrectomy for primary retinal detachment repair in presbyopes. AB - PURPOSE: To review the results of phacovitrectomy for primary retinal detachment repair in presbyopes. METHODS: The outcome and complications of surgery were examined in a retrospective case series of 93 presbyopic patients who underwent phacoemulsification and vitrectomy for primary rhegmatogenous retinal detachment. RESULTS: Eighty-two patients (88.2%) had reattachment with a single procedure. The final reattachment rate with further procedures was 97.8%. Two patients (2.2%) in whom final reattachment did not occur declined further surgery after the primary repair. All redetachments were due to proliferative vitreoretinopathy. The median Snellen converted logMAR visual acuity improved from 1.00 preoperatively to 0.30 postoperatively (P < 0.001, Wilcoxon test). Postoperative complications included transient intraocular pressure rise (29%), fibrinous uveitis (16.2%), and iris/intraocular lens capture (8.6%). One of the eight patients with iris/intraocular lens capture required surgical reintervention. All cases of fibrinous uveitis and intraocular pressure rise resolved within a few days with medical treatment. Posterior capsular opacification occurred in 23 patients (24.7%). CONCLUSION: Combined surgery with phacoemulsification, lens extraction, and vitrectomy offers significant advantages to both patient and surgeon in the management of primary retinal detachment. We believe that it should be considered for presbyopes even in the absence of significant lens opacity. Fibrinous uveitis and intraocular pressure rise may occur in a few patients in the immediate postoperative period but are transient and resolve with medical treatment. Further prospective studies are required, in particular to examine the rate of postoperative proliferative vitreoretinopathy, which may be higher than with vitrectomy alone. PMID- 17420700 TI - Prophylaxis for retinal detachment: evidence or eminence based? AB - PURPOSE: Prophylactic laser treatment of peripheral retinal lesions to prevent retinal detachment enjoys widespread use. However, clinical and scientific evidence for such treatment only exists for a few particular clinical situations. The aim of the present study was to delineate the current practice patterns for such prophylaxis among some European retinal specialists. METHODS: A postal survey was conducted among all members of BEAVRS (British and Eire Association of Vitreoretinal Surgeons; n = 236), all members of the German Retinological Society (n = 396), and Swiss retinal surgeons (n = 12) using a questionnaire with 6 case scenarios. The retinal specialists were asked to choose between three different options for each case: treat retinal lesion prophylactically, clinical follow-up, and other options. RESULTS: The return rates of the questionnaire were 36% in the United Kingdom, 42% in Germany, and 83% in Switzerland. There was a general tendency to apply laser prophylaxis to peripheral retinal lesions despite the absence of evidence in the medical literature. CONCLUSIONS: Nonevidence-based prophylactic laser treatment of peripheral retinal lesions remains a clinical and economic issue that must be addressed, and more evidence-based data are needed to establish clear clinical guidelines. Swiss retinal specialists are particularly conscientious in sending back a medical questionnaire. PMID- 17420701 TI - Full-thickness macular hole formation in idiopathic parafoveal telangiectasis. AB - PURPOSE: To describe the macular holes in patients with idiopathic parafoveal telangiectasis (IPT) and to propose a pathophysiologic explanation for their formation. METHODS: Four eyes of two patients with IPT were evaluated with biomicroscopy and optical coherence tomography (OCT). RESULTS: One patient had a nearly full-thickness hole with preservation of only the internal limiting membrane (ILM), but had a 20/60 visual acuity. The other patient had a large full thickness macular hole, but retained 20/40 visual acuity. Each patient had a fellow eye showing prominent central inner foveal cavitation under a very thin ILM, which was devoid of associated tissue. CONCLUSIONS: This report describes the findings of two patients with IPT who developed pronounced central foveal structural abnormalities. The induced anatomic changes noted in our patients suggest that there is a loss of the structural aspects afforded by Muller cells, particularly the Muller cell cone, in the central macula in patients with IPT. The preservation of good visual acuity in our patients implies that the holes were the result of lateral separation of the photoreceptors within the fovea and that there could not have been profound atrophy of the photoreceptors. PMID- 17420702 TI - Anatomical and visual outcomes of idiopathic macular hole surgery with internal limiting membrane removal using low-concentration indocyanine green. AB - PURPOSE: To report the anatomical, visual, and optical coherence tomography (OCT) results of using a brief application of a low concentration of indocyanine green (ICG) to assist the removal of internal limiting membrane (ILM) during idiopathic macular hole repair. METHODS: Retrospective, interventional, noncomparative case series of 59 eyes of 57 patients with stage 2, 3, or 4 idiopathic macular holes who underwent pars plana vitrectomy with removal of ILM assisted by a brief (<30 seconds) intravitreal application of 0.125% (1.25 mg/mL) ICG. RESULTS: The median follow-up period was 13 months (range, 2-40 months). Anatomical closure of the macular hole was achieved in 58 eyes (98%) with a single surgery. Visual acuity improved from a preoperative mean of 20/100 to 20/60 postoperatively (P < 0.0001). Twenty-nine eyes (49%) had postoperative visual acuity of 20/50 or better. Visual acuity improved by > or =2 lines in 43 eyes (73%) and between 0 and 2 lines in 13 eyes (22%) and decreased in 3 eyes (5%). Postoperative OCT showed closure of macular hole with normal foveal depression in 49 (89%) of 55 eyes. CONCLUSION: A brief application of ICG at a low concentration appears to provide a safe and effective way of assisting ILM peeling during idiopathic macular hole surgery. PMID- 17420703 TI - Effect of tinted optical filters on visual acuity and contrast sensitivity in patients with inflammatory cystoid macular edema. AB - PURPOSE: To ascertain the effect of tinted optical filters (TFs) on visual function in patients with inflammatory cystoid macular edema (CME). METHODS: A prospective study included 14 patients with inflammatory CME and best-corrected visual acuity between 0.1 and 0.8 (CME group) and 16 sex-, age-, and best corrected visual acuity-matched patients with macular disorders but no CME (non CME group). All patients from both groups underwent the following tests: determination of visual acuity at distance, contrast sensitivity (CS) testing; and subjective grading of each TF for best-corrected visual acuity and CS compared with no TFs. All tests were performed with the following TFs: Corning photochromic filter with a cutoff at 450 nm; Zeiss filter with a cutoff at 560 nm; and cheap widely available yellow filter with a cutoff at 489 nm. RESULTS: Without TFs, the CME and non-CME groups did not differ in best-corrected visual acuity (P = 0.79). Using the TFs, a clinically significant improvement was not achieved by any patient. However, using the definition of any improvement, best corrected visual acuity with all TFs showed a significant improvement in the CME group compared with no TFs (P < 0.01). The CME group rated the Corning photochromic filter with a cutoff of 450 nm significantly higher than the other TFs when determining visual acuity (P = 0.05) and CS (P = 0.01) in contrast to no improvement in the non-CME group. CONCLUSION: Although the use of TFs for the CME patients was not associated with criteria of clinically significant improvement, our results indicate that visual acuity and CS in patients with inflammatory CME might benefit from the use of TFs. PMID- 17420704 TI - Morphometric age-related evaluation of small retinal vessels by scanning laser Doppler flowmetry: determination of a vessel wall index. AB - BACKGROUND: Measurement of the retinal vessel wall thickness may contribute to the diagnosis of microvascular diseases. We present a methodical approach to calculate these alterations and to determine age-related differences. METHODS: One hundred fifty-three subjects without eye or internal diseases (mean age +/- SD, 47.6 +/- 14.9 years) underwent measurement of the retinal temporal superior artery and vein by scanning laser Doppler flowmetry (Heidelberg retina flowmeter). We calculated the difference between the diameter of reflectivity and the Doppler signal (Delta[VD-FD]/2) and determined a "vessel wall index" (VWI) by normalization of Delta(VD-FD)/2 for age and vessel diameter. RESULTS: Delta(VD FD)/2 correlated with vessel diameter (artery, r = +0.60, P < 0.001; vein, r = +0.49, P<0.001) and age (artery, r = +0.19, P = 0.02; vein, r = +0.27, P = 0.001) but not with sex, if controlled for the other variables each. The venous, but not the arterial, vessel diameter correlated with age (r = +0.18, P = 0.02), if controlled for sex. The relative statistical weight of these empirical contributions to the variation observed in Delta(VD-FD)/2 was 36.5% (P < 0.001, artery) and 21.7% (P< 0.001, vein), and that of age was 3.6% (P = 0.02, artery) and 7.3% (P = 0.001, vein). The limit value of VWI to pathologic changes (80th percentile) was 1.25 microm/y (artery) and 1.31 microm/y (vein). Delta(VD-FD)/2 normalized for vessel diameter correlated with the 10-year categories of age (artery, r = +0.196, P = 0.017; vein, r = +0.250, P = 0.002). CONCLUSION: In a group of subjects aged 21 years to 70 years, we detected an increase of Delta(VD FD)/2 in the retinal temporal superior artery and vein with age. PMID- 17420705 TI - Biocompatibility of brilliant blue G in a rat model of subretinal injection. AB - PURPOSE: To evaluate the toxicity of brilliant blue G (BBG) compared with those of indocyanine green (ICG) and trypan blue (TB) in a rat model of subretinal injection. METHODS: Retinal detachment was produced by subretinal injection of the dyes. The biocompatibility of BBG (0.25 mg/mL) was evaluated over 2 months and 2 weeks by ophthalmic examinations. The eyes were enucleated and analyzed by light, fluorescence, as well as transmission electron microscopy. Apoptotic cell death was detected by TdT-dUTP terminal nick-end labeling. The results were compared with those for ICG (5 mg/mL) and TB (1 mg/mL). RESULTS: ICG caused retinal degeneration and retinal pigment epithelial (RPE) cell atrophy 2 weeks after subretinal injection. Apoptotic cell death was detected in the inner and outer nuclear layers and the RPE layer, especially the photoreceptors. TB caused less retinal degeneration, mainly in the area detached by the subretinal injection. BBG had no detectable toxic effects after 2 months and 2 weeks. Apoptotic cell death was detected in the ICG and TB groups, mainly in the photoreceptors. CONCLUSIONS: Subretinal injection of the dyes caused retinal cell degeneration at lower concentrations than those reported for intravitreous injection. However, subretinal injection of BBG at 0.25 mg/mL appeared to provide satisfactory biocompatibility. PMID- 17420706 TI - Repair of a traumatic scleral rupture with scleral imbrication and BioGlue. PMID- 17420707 TI - Diagnostic and therapeutic challenges. PMID- 17420709 TI - Venous occlusive disease review. PMID- 17420710 TI - Molecular imaging. PMID- 17420711 TI - The uncertain path to the future of imaging biomarkers. PMID- 17420712 TI - Nuclear medicine applications in molecular imaging: 2007 update. AB - This review examines several classes of radiolabeled agents, including analogs localizing in somatostatin, benzodiazepine and dopamine receptors; analogs of progesterone and estrogen; and agents localizing in lesions with hypoxia. It concludes the status of agents advocated for detecting angiogenesis and inflammation. The current clinical status of these agents, and their potential roles in diagnosis and treatment are discussed. PMID- 17420713 TI - Instrumentation and probes for molecular and cellular imaging. AB - Molecular and cellular imaging is a branch of biomedical sciences that combines the use of imaging instrumentation and biotechnology to characterize molecular and cellular processes in living organisms in normal and pathologic conditions. The two merging areas of research behind molecular and cellular imaging are detection technology, i.e. scanners and imaging devices, and development of tracers, contrast agents and reporter probes that make imaging with scanners and devices possible. Several in vivo imaging instruments currently used in human studies, such as computer tomography, ultrasound, magnetic resonance, positron emission tomography and single photon emission computed tomography, have been rescaled for small animal studies, while other methods initially used for in vitro evaluation, such as bioluminescence and fluorescence, have been refined for in vivo imaging. Conventional imaging relies on the use of non specific contrast agents and classical probes; however, newly developed targeted contrast agents and activable ''smart'' imaging probes for so-called ''targeted imaging'' have demonstrated high specificity and high signal to noise ratio in small animal studies. This review focuses on basic recent findings in the technical aspects of molecular and cellular imaging modalities (equipment, targeted probe and contrast agents and applied combinations of instrumentation and probe) with particular attention to the choice of the future: the multimodal imaging approach. PMID- 17420714 TI - Multimodality imaging: novel pharmacological applications of reporter systems. AB - The development of novel drugs is a lengthy process requiring years of preclinical research and many steps indispensable to ensure that the molecule of interest can be administered to humans with a minimal risk of toxic effects. Even a minimal reduction in the initial stages of drug development would result in a tremendous saving in time; therefore, pharmaceutical companies are eager to apply novel methodologies that shorten the time required for pharmacodynamic, pharmacokinetic and toxicological studies to be carried out in vitro and in animal systems. Currently, quantitative analysis of molecular events in living organisms is done with the combined application of imaging and genetic engineering technologies. In vivo imaging provides surrogate endpoints that can improve the identification of new drug candidates and speed up their research at preclinical stages. The integration of reporter systems in animal models of human diseases represents a reachable frontier that will dramatically advance drug development in terms of costs, time and efficacy. The present review outlines the applicability of imaging technologies for drug development and presents a panorama on the state of the art of currently available imaging technologies suitable for preclinical studies, with particular focus on bioluminescence and fluorescence as the methodologies of election. PMID- 17420715 TI - Radiolabelled chemotherapeutics. AB - During the past 3 decades various chemotherapeutics have been directly or indirectly radiolabeled for single photon emission computed tomography (SPECT) and positron emission tomography (PET) imaging. Several of these radiolabeled chemotherapeutics have been injected into patients for the purpose of better understanding their biodistribution and metabolism and to assess whether there exists a relationship between their uptake in tumor tissue and response to treatment. Based on available data, it may be concluded that PET or SPECT imaging with radiolabeled chemotherapeutics provides valuable information that helps to better understand the mechanism of action and the metabolic conversion of unlabeled chemotherapeutics in humans and that helps to provide a rationale for the lack of response to certain chemotherapeutics or for the beneficial effect of biomodulating agents. As for their utility as predictors of response to therapy, their utility is limited to those agents given in monotherapy. PMID- 17420716 TI - Nuclear imaging in cancer theranostics. AB - The combination of a diagnostic test with a therapeutic entity is termed theranostics. The diagnostic test aims at identifying patients who will likely benefit from a specific therapeutic intervention, fail to respond or eventually manifest side effects to a given drug. The appropriate selection of patients for innovative therapies would promote an enrichment of patient population that can potentiate clinical trials and, eventually, accelerate the drug development process. For these reasons, many drug companies adopted a theranostic approach as a new and promising avenue for drug development. From an historical perspective, the concepts underlying the theranostic strategy are well known in nuclear medicine and constituted the basis of many nuclear imaging procedures currently used in clinical practice. Nevertheless the adoption of these concepts by regulatory authorities is a real progress and reflects the remarkable advances in the development of drugs against molecular targets. In this respect, the oncological field provides the strongest evidence of the clinical need to link diagnostics to therapeutics. Here, we review the contribution that non-invasive nuclear imaging may give to cancer theranostics and report prominent examples of nuclear imaging procedures that can be coupled to specific therapies. The main focus lies on imaging procedures that can identify patients who will benefit from molecularly targeted therapy or are going to fail to respond to standard treatment. PMID- 17420717 TI - Role of in vivo imaging of the central nervous system for developing novel drugs. AB - Over the past decade imaging technologies employed in clinical neurosciences have significantly advanced. Imaging is not only used for the diagnostic work-up of neurological disorders but also crucial to follow up on therapeutic efforts. Using disease-specific imaging parameters, as read-outs for the efficiency of individual therapies, has facilitated the development of various novel treatments for neurological disease. Here, we review various imaging technologies, such as cranial computed tomography (CT), magnetic resonance imaging (MRI) and spectroscopy (MRS), positron emission tomography (PET) and single-photon emission computed tomography (SPECT), with respect to their current applications in non invasive disease phenotyping and the measurement of therapeutic outcomes in neurology. In particular, applications in neuro-oncology, Parkinson's disease, Alzheimer's disease, and cerebral ischemia are discussed. Non-invasive imaging provides further insights into the molecular pathophysiology of human diseases and facilitates the design and implementation of improved therapies. PMID- 17420718 TI - Failure predictors of the sentinel lymph node in patients with breast cancer using Tc-99m sulfur colloid and periareolar injection. AB - AIM: As sentinel lymph node (SLN) experience rises, it is important to identify factors that can limit lymphoscintigraphic mapping. METHODS: A prospective study was conducted with breast cancer patients that were submitted to sentinel node mapping by lymphoscintigraphy between October 2003 and January 2005. The analyzed factors were: patients' age, body mass index, tumor size, previous breast surgeries, time between a previous biopsy and the radiotracer injection and their impact on preoperative SLN identification. RESULTS: Two hundred and three breast cancer patients were injected with (99m)Technetium-sulfur colloid and submitted to lymphoscintigraphy scan for SLN biopsy. One hundred and eighty-four of these patients (90.64%) had a successfully identified SLN and 19 (9.36%) had a mapping failure. The median age of the successful group was 55.6 years and in the failure group was 57.1 years (P=0.002). The median body mass index was 25.3 and 27.6, respectively (P=0.024). The tumor size did not show any significant difference between the patients with successful mapping and failure (P=0.07). Previous breast surgery was an important limiting factor for SLN mapping (P=0.017). The mean time from biopsy to SLN detection was 23.6 days on the successfully marked patients and 17.4 days in the failure group (P<0.0001). All the 184 successfully mapped patients had the SLN identified. Only one patient of the failure group had the SLN identified using blue dye. CONCLUSION: Advanced age, elevated body mass index, previous breast surgery and a shorter period of time after a breast biopsy are causes for SLN identification failure. The tumor size was not a limiting factor. PMID- 17420719 TI - High resolution small animal single photon emission computed tomography: uptake of [99mTc]bombesin and [123I]ioflupane by rat brain. AB - AIM: The aims of this study were: 1) to perform brain single photon emission computed tomography (SPECT) in anesthetized rats with high resolution cameras (HRC) equipped with parallel hole collimation resolution of about 1 mm (HRC1) and 2 mm (HRC2); 2) to assess when and with which radio-tracer HRC1 SPECT shows advantages over HRC2. METHODS: We used two multicrystal HRCs with parallel square hole collimators, whose pure tungsten septa closely fit the crystals, in turn matched with a 4 inch2 position sensitive photomultiplier. HRC1 showed 1.1 mm and HCR2 2.1 mm resolution at collimator contact. HRCs performed 180 degrees semi circular orbits around the head of rats: image reconstruction occurred with ordered subsets expectation maximization algorithms. Resolution of SPECT was measured with a Derenzo Phantom, resulting 1.4 mm for HRC1 and 2.3 mm for HRC2. Three rats were studied with [(99m)Tc]HMPAO, 3 rats with [(99m)Tc]bombesin (BN) and 48 h later with [(123)I]ioflupane (DaTSCAN). SPECT studies were reviewed by two experienced operators. RESULTS: Technetium-99m-HMPAO SPECT showed similar images with HRC1 and HRC2. The uptake of BN by amygdale, hippocampus and olfactory tract was detected by both cameras. DaTSCAN SPECT with HRC1 showed detailed image of the tail of the caudatus: this image was not obtained with HRC2. DaTSCAN and BN SPECT showed amygdale with both HRCs. However, only the central nucleus of amygdale takes up DaTSCAN, whereas central, lateral and basolateral amygdaloid nuclei express BN receptors. Only HRC1 SPECT showed amygdale larger with BN than with DaTSCAN. CONCLUSION: Spatial resolution of 1.4 mm is appropriate to detect selected subcerebral structures. PMID- 17420720 TI - Brca1 heterozygous mice have shortened life span and are prone to ovarian tumorigenesis with haploinsufficiency upon ionizing irradiation. AB - BRCA1 mutation carriers have an 85% lifetime risk of breast cancer and 60% for ovarian cancer. BRCA1 facilitates DNA double-strand break repair, and dysfunction of BRCA1 leads to hypersensitivity to DNA damaging agents and consequently genomic instability of cells. In this communication, we have examined the tumor incidence and survival of Brca1 heterozygous female mice. Brca1 heterozygotes appear to have a shortened life span with 70% tumor incidence. Lymphoma, but not ovarian and mammary gland tumors, occurs commonly in these mice. After a whole body exposure to ionizing radiation, Brca1 heterozygous mice have a 3-5-fold higher incidence specific to ovarian tumors, but not lymphoma, when compared with the Brca1+/+ mice. All the tumors from heterozygous mice examined retain the wild type allele and the cancer cells express Brca1 protein, precluding the chromosomal mechanism for loss of heterozygosity of Brca1 locus. Although the manifestation of BRCA1 haploinsufficiency may be different between human and mouse, this study suggests that women carrying Brca1 mutations may be more prone to ovarian tumor formation after IR exposure than nonmutation carriers. PMID- 17420721 TI - Siah-1S, a novel splice variant of Siah-1 (seven in absentia homolog), counteracts Siah-1-mediated downregulation of beta-catenin. AB - Siah-1 (seven in absentia homolog) is known to cause indirect degradation of beta catenin through formation of a complex with Siah-interacting protein (SIP), Skp1 and Ebi. Here, we report the characterization of a novel splice variant of human Siah-1, designated Siah-1S, which is produced by an alternative splicing mechanism. The novel intron/exon junctions used to generate Siah-1S follow a non conventional CT-AC rule. Siah-1S exhibits an even shorter half-life than Siah-1 and is able to catalyse self-ubiquitination that results in its subsequent degradation by proteasome. Siah-1S is shown to upregulate beta-catenin-dependent Tcf/Lef transcriptional activation and antagonize Siah-1's potentiation effect on the apoptosis induced by etoposide in MCF-7 cells. Additionally, Siah-1S is found to interact with Siah-1 to form heterodimer or with itself to form homodimer. Unlike homodimer Siah-1*Siah-1, neither Siah-1*Siah-1S nor Siah-1S*Siah-1S is able to bind to Siah-1-interacting protein, which may explain the underlying mechanism for Siah-1S's dominant negative effect on Siah-1. Importantly, results from in vitro soft agar assay demonstrated that Siah-1S displays a promotion effect on cells tumorigenicity. PMID- 17420722 TI - Rituximab inhibits the constitutively activated PI3K-Akt pathway in B-NHL cell lines: involvement in chemosensitization to drug-induced apoptosis. AB - Rituximab (chimeric anti-CD20 monoclonal antibody) is currently being used, alone or in combination with chemotherapy, in the treatment of B-non-Hodgkin's lymphoma (B-NHL). We have reported that rituximab treatment of B-NHL cell lines sensitizes the drug-resistant tumor cells to apoptosis by various chemotherapeutic drugs and chemosensitization was, in large part, owing to the selective inhibition of the anti-apoptotic Bcl-(XL) gene product. The constitutive activation of the Akt pathway in B-NHL results in overexpression and functional activation of Bcl-(xL). Hence, we hypothesized that rituximab-induced inhibition of Bcl-(xL) expression and chemosensitization may result, in part, from its inhibitory activity of the Akt pathway. This hypothesis was tested using the drug-resistant Ramos and Daudi B-NHL cell lines. Time kinetic analysis revealed that treatment with rituximab inhibited phosphorylation of Akt, but not unphosphorylated Akt, and the inhibition was first detected at 6 h post-rituximab treatment. Similar time kinetics revealed rituximab-induced inhibition of p-PDK1, p-Bad, p-IKKalpha/beta and p-Ikappabetaalpha and no inhibition of unphosphorylated proteins. In addition, rituximab treatment resulted in significant increase of Bcl-(xL)-Bad heterodimeric complexes as compared to untreated cells. The role of the Akt pathway in the regulation of resistance was corroborated by the use of the Akt inhibitor, LY294002, and by transfection with siRNA Akt. Treatment of tumor cells with LY294002 or with Akt siRNA, but not control siRNA, resulted in inhibition of Bcl-(xL) expression and sensitization to drug-induced apoptosis. Although rituximab did not inhibit the Akt pathway nor sensitized the rituximab-resistant Ramos RR1 clone, treatment with LY294002 or Akt siRNA sensitized the clone to drug-induced apoptosis. The present findings demonstrate for the first time that rituximab inhibits the constitutively activated Akt pathway in B-NHL cell lines, and this inhibition contributes to sensitization of drug-resistant cells to apoptosis by chemotherapeutic drugs. The findings also identify the Akt pathway as target for therapeutic intervention in the reversal of rituximab and drug resistant B-NHL. PMID- 17420723 TI - Involvement of H4(D10S170) protein in ATM-dependent response to DNA damage. AB - H4(D10S170) gene has been identified upon its frequent rearrangement with RET in papillary thyroid tumours (RET/PTC1). The kinase ataxia telangectasia mutated (ATM) phosphorylates a limited number of downstream protein targets in response to DNA damage. We investigated the potential role of H4(D10S170) in DNA damage signaling pathways. We found that in cells treated with etoposide or ionizing radiation (IR), H4(D10S170) underwent ATM-mediated phosphorylation at Thr 434, stabilizing nuclear H4. In ataxia telangectasia cells (A-T), endogenous H4(D10S170) was localized to cytoplasm and was excluded from the nucleus. Moreover, H4(D10S170) was not phosphorylated in ATM-deficient lymphoblasts after ionizing irradiation. Inhibition of ATM kinase interfered with H4(D10S170) apoptotic activity, and expression of H4 with threonine 434 mutated in Alanine, H4(T434A), protected the cells from genotoxic stress-induced apoptosis. Most importantly, after exposure to IR we found that silencing of H4(D10S170) in mammalian cells increased cell survival, as shown by clonogenic assay, allows for DNA synthesis as evaluated by bromodeoxyuridine incorporation and permits cells to progress into mitosis as demonstrated by phosphorylation on Histone H3. Our results suggest that H4(D10S170) is involved in cellular response to DNA damage ATM-mediated, and that the impairment of H4(D10S170) gene function might have a role in thyroid carcinogenesis. PMID- 17420724 TI - p21 provides stage specific DNA damage control to preimplantation embryos. AB - The early stage embryogenesis of higher eukaryotes lacks some of the damage response pathways such as G1/S checkpoint, G2/M checkpoint and apoptosis. We examined here the damage response of preimplantation stage embryos after fertilization with 6 Gy irradiated sperm. Sperm-irradiated embryos developed normally for the first 2.5 days, but started to exhibit a developmental delay at day 3.5. p21 was activated in the delayed embryos, which carried numerous micronuclei owing to delayed chromosome instability. Apoptosis was observed predominantly in the inner cell mass of the day 4.0 embryos. Sperm-irradiated p21 /- embryos lacked the delay, but chromosome instability and apoptosis were more pronounced than the corresponding p21 wild-type embryos. We conclude from the result that damage responses come in a stage-specific manner during preimplantation stage development; p53-dependent S checkpoint at the zygote stage, p21-mediated cell cycle arrest at the morula/blastocyst stages and apoptosis after the blastocyst stage in the inner cell mass. PMID- 17420725 TI - A 90 kDa fragment of filamin A promotes Casodex-induced growth inhibition in Casodex-resistant androgen receptor positive C4-2 prostate cancer cells. AB - Prostate tumors are initially dependent on androgens for growth, but the majority of patients treated with anti-androgen therapy progress to androgen-independence characterized by resistance to such treatment. This study investigates a novel role for filamin A (FlnA), a 280 kDa cytoskeletal protein (consisting of an actin binding domain (ABD) followed by 24 sequential repeats), in androgen-independent (AI) growth. Full-length FlnA is cleaved to 170 kDa (ABD+FlnA1-15) and 110 kDa fragments (FlnA16-24); the latter is further cleaved to a 90 kDa fragment (repeats 16-23) capable of nuclear translocation and androgen receptor (AR) binding. Here, we demonstrate that in androgen-dependent LNCaP prostate cancer cells, the cleaved 90 kDa fragment is localized to the nucleus, whereas in its AI subline C4-2, FlnA failed to cleave and remained cytoplasmic. Transfection of FlnA16-24 cDNA in C4-2 cells restored expression and nuclear localization of 90 kDa FlnA. Unlike LNCaP, C4-2 cells proliferate in androgen-reduced medium and in the presence of the AR-antagonist Casodex. They also exhibit increased Akt phosphorylation compared to LNCaP, which may contribute to their AI phenotype. Nuclear expression of 90 kDa FlnA in C4-2 cells decreased Akt phosphorylation, prevented proliferation in androgen-reduced medium and restored Casodex sensitivity. This effect was inhibited by constitutive activation of Akt indicating that FlnA restored Casodex sensitivity in C4-2 cells by decreasing Akt phosphorylation. In addition, FlnA-specific siRNA which depleted FlnA levels, but not control siRNA, induced resistance to Casodex in LNCaP cells. Our results demonstrate that expression of nuclear FlnA is necessary for androgen dependence in these cells. PMID- 17420726 TI - Downregulation of Sef, an inhibitor of receptor tyrosine kinase signaling, is common to a variety of human carcinomas. AB - Carcinomas are tumors of epithelial origin accounting for over 80% of all human malignancies. A substantial body of evidence implicates oncogenic signaling by receptor tyrosine kinases (RTKs) in carcinoma development. Here we investigated the expression of Sef, a novel inhibitor of RTK signaling, in normal human epithelial tissues and derived malignancies. Human Sef (hSef) was highly expressed in normal epithelial cells of breast, prostate, thyroid gland and the ovarian surface. By comparison, substantial downregulation of hSef expression was observed in the majority of tumors originating from these epithelia. Among 186 primary carcinomas surveyed by RNA in situ hybridization, hSef expression was undetectable in 116 cases including 72/99 (73%) breast, 11/16 (69%) thyroid, 16/31 (52%) prostate and 17/40 (43%) ovarian carcinomas. Moderate reduction of expression was observed in 17/186, and marked reduction in 40/186 tumors. Only 13/186 cases including 12 low-grade and one intermediate grade tumor retained high hSef expression. The association of hSef downregulation and tumor progression was statistically significant (P<0.001). Functionally, ectopic expression of hSef suppressed proliferation of breast carcinoma cells, whereas inhibition of endogenous hSef expression accelerated fibroblast growth factor and epidermal growth factor-dependent proliferation of cervical carcinoma cells. The inhibitory effect of hSef on cell proliferation combined with consistent downregulation in human carcinoma indicates a tumor suppressor-like role for hSef, and implicates loss of hSef expression as a common mechanism in epithelial neoplasia. PMID- 17420727 TI - Sacrificing the cash cow. PMID- 17420728 TI - Pharma consolidates its grip on post-antibody landscape. PMID- 17420729 TI - US courts thwart GM alfalfa and turf grass. PMID- 17420730 TI - UK parses merits of value-based drug pricing. PMID- 17420731 TI - Genentech faces the consequences of growth. PMID- 17420733 TI - Profile: John Maraganore. Interviewed by Brady Huggett. PMID- 17420734 TI - Making money and opening minds. PMID- 17420735 TI - Billion dollar babies--biotech drugs as blockbusters. PMID- 17420736 TI - WARF's licensing policy for ES cell lines. PMID- 17420737 TI - Gene patents--more evidence needed, but policymakers must act. PMID- 17420738 TI - Nanobiotech versus synthetic nanotech? PMID- 17420739 TI - Systems biology standards--the community speaks. PMID- 17420740 TI - Incentivizing standards development and adoption. PMID- 17420741 TI - Commercializing cognitive neurotechnology--the ethical terrain. PMID- 17420742 TI - Research sharing, ethics and public benefit. PMID- 17420744 TI - India's health biotech sector at a crossroads. PMID- 17420745 TI - The global stem cell patent landscape: implications for efficient technology transfer and commercial development. PMID- 17420746 TI - Recent patent applications in proteomics. PMID- 17420747 TI - Towards universal red blood cells. PMID- 17420748 TI - Antibiotic production from the ground up. PMID- 17420749 TI - Tagging the rice transcriptome. PMID- 17420750 TI - Vascular targeting via caveolae. PMID- 17420751 TI - Lessons from the 1918 influenza. PMID- 17420760 TI - Managing change in biotech: startup and growth. PMID- 17420753 TI - Overhauling the PDB. PMID- 17420762 TI - Apolipoprotein E polymorphism, homocysteine serum levels and hippocampal volume in patients with alcoholism: an investigation of a gene-environment interaction. AB - There is growing evidence that disadvantageous influences of the apolipoprotein E4 allele in the central nervous system are modified by environmental and dietary conditions. The present study investigated the gene-environment interaction of apolipoprotein E4 with homocysteine serum levels in patients with alcohol dependence with regard to alcohol-related hippocampal volume loss using volumetric high-resolution magnetic resonance imaging. We included 52 patients with alcohol-dependence. ApoE genotypes, homocysteine serum levels and hippocampal volumes were determined. We found a significant impact of homocysteine (F=13.2; df=1; P<0.001; 1-beta=0.95), not for ApoE4 genotype (F=0.482; df=1; P=0.49; 1-beta=0.05) on hippocampal volume. There was a significant interaction between both factors (ApoE4 x Hcy; F=8.8; df=1; P=0.005; 1-beta=0.80). The ApoE4 allele constitutes a risk factor for hippocampal volume loss in patients with alcohol dependence under the conditions of hyperhomocysteinemia. We suggest that the disadvantageous effects of apolipoprotein E4 on alcohol-related brain volume loss are based on certain gene environment interactions. PMID- 17420763 TI - Parasagittal biopsies are more important as part of an initial biopsy strategy than as part of a repeat biopsy strategy: observations from a unique population. AB - Comparing the yield of parasagittal biopsies during initial saturation biopsy to the yield during repeat saturation biopsy for detection of prostate cancer. Office-based saturation biopsy (24 cores) with periprostatic lidocaine block was performed in 139 consecutive men who had never previously undergone prostate biopsy. Indication for biopsy was elevated prostate-specific antigen >2.5 ng/dl. Biopsy specimens were obtained and marked by location for histological examination. Subanalysis of patients from this unique study was performed to compare the location of saturation biopsy cancer detection in these patients to a cohort of 100 patients who had previously undergone biopsy with nonmalignant findings. In the initial biopsy group, cancer was detected in 62/139 patients (44.6%). Breakdown of cancer location demonstrated unique parasagittal cancers in 9/62 patients (14.5%). Laterally base cancer was found exclusively in 22/62 patients (35.5%). For the repeat biopsy population, cancer was found in 25 patients (25%); no patients (0%) had exclusive parasagittal cancer. To our knowledge, this is the first study to demonstrate a difference in the location of positive cores between initial and repeat biopsy status. The exclusive parasagittal cancer detection rate decreases significantly in the repeat biopsy population when using the same biopsy method. Our findings support including traditional template parasagittal sampling of the prostate on first-time biopsy in addition to lateral cores typical of extended field biopsies for a total of 10 12 cores. However, parasagittal sampling adds negligible additional information in repeat biopsy; thus we recommend obtaining primarily laterally based cores for repeat biopsy. PMID- 17420764 TI - Immunology and immunotherapy approaches for prostate cancer. AB - Several mechanisms that impair the immune response to promote tumour progression are reported. These mechanisms aim to reduce the ability of antigen-presenting cells to present antigen and activate naive T cells to support an active immune response or to create a suppressive environment that induce non-functional tumour associated antigen-specific T cells. Prostate cancer (PC) alone accounts for 33% of incident cancer cases and about 9% of all cancer-related deaths among men in the USA during 2006. Whereas androgen deprivation has remained the first line of therapy for advanced PC, other therapies are still required due to progression to an androgen-resistant state and eventually loss of control in patients receiving hormonal therapy. Immunotherapy seems to be a promising approach to enhance tumour-specific T-cell responses in different cancers including prostate. More importantly, clinical trials in advanced PC patients have shown that immunotherapy may generate significant clinical responses. Immunology and immunotherapy aspects of PC with focus on prostate-specific antigen will be presented. PMID- 17420765 TI - Molecular studies of major depressive disorder: the epigenetic perspective. AB - Major depressive disorder (MDD) is a common and highly heterogeneous psychiatric disorder encompassing a spectrum of symptoms involving deficits to a range of cognitive, psychomotor and emotional processes. As is the norm for aetiological studies into the majority of psychiatric phenotypes, particular focus has fallen on the interplay between genetic and environmental factors. There are, however, several epidemiological, clinical and molecular peculiarities associated with MDD that are hard to explain using traditional gene- and environment-based approaches. Our goal in this study is to demonstrate the benefits of looking beyond conventional 'DNA+environment' and 'DNA x environment' aetiological paradigms. Epigenetic factors - inherited and acquired modifications of DNA and histones that regulate various genomic functions occurring without a change in nuclear DNA sequence - offer new insights about many of the non-Mendelian features of major depression, and provide a direct mechanistic route via which the environment can interact with the genome. The study of epigenetics, especially in complex diseases, is a relatively new field of research, and optimal laboratory techniques and analysis methods are still being developed. Incorporating epigenetic research into aetiological studies of MDD thus presents a number of methodological and interpretive challenges that need to be addressed. Despite these difficulties, the study of DNA methylation and histone modifications has the potential to transform our understanding about the molecular aetiology of complex diseases. PMID- 17420767 TI - HIV transmission: migratory Langerhans cells are primary targets in vaginal HIV transmission. PMID- 17420766 TI - Lack of serotonin1B receptor expression leads to age-related motor dysfunction, early onset of brain molecular aging and reduced longevity. AB - Normal aging of the brain differs from pathological conditions and is associated with increased risk for psychiatric and neurological disorders. In addition to its role in the etiology and treatment of mood disorders, altered serotonin (5 HT) signaling is considered a contributing factor to aging; however, no causative role has been identified in aging. We hypothesized that a deregulation of the 5 HT system would reveal its contribution to age-related processes and investigated behavioral and molecular changes throughout adult life in mice lacking the regulatory presynaptic 5-HT(1B) receptor (5-HT(1B)R), a candidate gene for 5-HT mediated age-related functions. We show that the lack of 5-HT(1B)R (Htr1b(KO) mice) induced an early age-related motor decline and resulted in decreased longevity. Analysis of life-long transcriptome changes revealed an early and global shift of the gene expression signature of aging in the brain of Htr1b(KO) mice. Moreover, molecular changes reached an apparent maximum effect at 18-months in Htr1b(KO) mice, corresponding to the onset of early death in that group. A comparative analysis with our previous characterization of aging in the human brain revealed a phylogenetic conservation of age-effect from mice to humans, and confirmed the early onset of molecular aging in Htr1b(KO) mice. Potential mechanisms appear independent of known central mechanisms (Bdnf, inflammation), but may include interactions with previously identified age-related systems (IGF 1, sirtuins). In summary, our findings suggest that the onset of age-related events can be influenced by altered 5-HT function, thus identifying 5-HT as a modulator of brain aging, and suggesting age-related consequences to chronic manipulation of 5-HT. PMID- 17420768 TI - Spatial modelling of brief and long interactions between T cells and dendritic cells. AB - In the early phases of an immune response, T cells of appropriate antigen specificity become activated by antigen-presenting cells in secondary lymphoid organs. Two-photon microscopy imaging experiments have shown that this stimulation occurs in distinct stages during which T cells exhibit different motilities and interactions with dendritic cells (DCs). In this paper, we utilize the Cellular Potts Model, a model formalism that takes cell shapes and cellular interactions explicitly into account, to simulate the dynamics of, and interactions between, T cells and DCs in the lymph node paracortex. Our three dimensional simulations suggest that the initial decrease in T-cell motility after antigen appearance is due to "stop signals" transmitted by activated DCs to T cells. The long-lived interactions that occur at a later stage can only be explained by the presence of both stop signals and a high adhesion between specific T cells and antigen-bearing DCs. Furthermore, our results indicate that long-lasting contacts with T cells are promoted when DCs retract dendrites that detect a specific contact at lower velocities than other dendrites. Finally, by performing long simulations (after prior fitting to short time scale data) we are able to provide an estimate of the average contact duration between T cells and DCs. PMID- 17420769 TI - Mathematical models of cytotoxic T-lymphocyte killing. AB - By killing infected host cells, cytotoxic T lymphocytes (CTL) mediate an important defense mechanism against viruses and other intracellular pathogens. Quantitative aspects of this killing process have been studied for several decades in vitro. More recently, methods have been developed to measure the timescales of CTL killing in vivo. Here, we review the estimates of kinetic rates involved in CTL killing which were obtained in these studies, and elaborate on the differences between them. PMID- 17420770 TI - The correlation between nitric oxide and vascular endothelial growth factor in spinal cord injury. AB - STUDY DESIGN: Prospective, randomized, placebo-controlled, experimental study. OBJECTIVES: The issue of whether nitric oxide (NO) production is beneficial or deleterious on ischemic injuries of the central nervous system still remains doubtful. Vascular endothelial growth factor (VEGF) is known to induce the release of NO from endothelial cells. However, the effect of NO on VEGF synthesis is not clear. We aimed to determine the effects of L-arginine and NG-nitro-L arginine methyl ester (L-NAME) on VEGF synthesis and free radicals in a rat model of spinal cord ischemia-reperfusion (IR) injury. SETTING: Surgical Research Laboratory of a Medical School. MATERIAL AND METHODS: Twenty-eight Wistar rats were divided into four groups as follows (n=7): Sham, IR injury, L-arginine, and L-NAME. Infrarenal abdominal aorta was occluded to induce spinal cord ischemia. L Arginine (100 mg/kg) and L-NAME (10 mg/kg) were given before aortic occlusion. Biochemical assays of malondialdehyde (MDA), NO and VEGF were carried out in spinal cord specimens. RESULTS: L-Arginine treatment significantly increased MDA and NO, but decreased VEGF levels in spinal cord. However, nonselective inhibition of NOS with L-NAME significantly decreased MDA and NO, but increased VEGF levels. Besides, the positive linear correlation between MDA and NO, and negative linear correlations between MDA, NO and VEGF levels have also been demonstrated. CONCLUSION: Nonselective inhibition of NO synthase activity with L NAME attenuated free radical formation and increased VEGF level when compared with NO precursor L-arginine in a rat model of spinal cord ischemia. We suggest that inhibition of NO synthase, as well as induction of VEGF, may be a therapeutic option in spinal cord IR injury. PMID- 17420771 TI - Laparoscopic endopelvic sacral implantation of a Brindley controller for recovery of bladder function in a paralyzed patient. AB - BACKGROUND: A number of techniques are being investigated to accomplish bladder control recovery in paralyzed patients using the neurostimulation, but currently, all techniques are based on the dorsal implantation of the electrodes using a laminectomy. METHODS: On 27 April 2006 we performed a laparoscopic implantation of a Finetech-Brindley bladder controller on the endopelvic sacral roots in a Th8 completely paralyzed woman who had previously undergone the removal of a Brindley controller due to an arachnoiditis after extrathecal implantation with intradural sacral deafferentation. RESULTS: We required about 3.5 h for the entire surgical procedure; no complications occurred and the patients went home on 5th postoperative day. The patient is now able to void empty her bladder and her rectum using the controller without further need for self-catheterisation. CONCLUSIONS: The presented new technique of laparoscopic implantation of electrodes on the endopelvic portion of the sacral nerve roots is an option to be considered in all paralyzed patients with further wish for electrical induced miction/defecation after previous deafferentation. PMID- 17420772 TI - Orthostatic hypotension in the first month following acute spinal cord injury. AB - STUDY DESIGN: Retrospective data analysis. OBJECTIVES: To determine prevalence of orthostatic hypotension (OH) in patients with spinal cord injury (SCI) during the acute rehabilitation period. SETTING: Quaternary care spinal unit, Vancouver General Hospital, British Columbia, Canada. METHODS: Eighty-nine patients with acute SCI stratified by neurological level (cervical, 55 (62%); upper thoracic, 12 (13%); lower thoracic, 22 (25%)), and graded by American Spinal Injury Association standards. Non-invasive measurement of systolic and diastolic blood pressure and heart rate were made at baseline and 3 min following an orthostatic challenge test administered during the first month after SCI. RESULTS: Patients with cervical or upper thoracic motor complete SCI more frequently experienced OH (P<0.01). OH persisted during the first month following SCI in 74% of cervical and only 20% of upper thoracic motor complete SCI patients. CONCLUSION: Patients with cervical and upper thoracic motor complete SCI are more likely to experience persistent OH than those with lower level or motor incomplete SCI during the first month of rehabilitation. PMID- 17420773 TI - Transplantation of porous tubes following spinal cord transection improves hindlimb function in the rat. AB - STUDY DESIGN: Experimental. OBJECTIVE: To determine the effects of a porous tube transplant in spinal cord transected rats. SETTING: Acadia University, Wolfville, Nova Scotia, Canada. METHODS: Female rats were randomly assigned to three experimental groups: control (Con, n=8), spinal cord transected (Tx, n=5) and spinal cord transected with transplant (TxTp, n=7). The rats in the TxTp and Tx groups received a complete spinal cord transection at the T10 level and the TxTp group immediately received a porous tube transplant. RESULTS: Locomotor activity rated on the Basso, Beattie, Bresnahan scale improved significantly in the TxTp animals over the 4 weeks such that final scores were 21, 1.4 and 7.1 for the Con, Tx and TxTp groups, respectively. As expected, the muscle to body mass ratios of the hindlimb skeletal muscles of the Tx group were decreased (soleus 35%, plantaris 29% and gastrocnemius 29%) and this was also observed in the TxTp group (soleus 33%, plantaris 23% and gastrocnemius 30%). Cytochrome c oxidase (CYTOX) activity in the plantaris was decreased by Tx but maintained in the TxTp group (Con=82.2, Tx=44.8 and TxTp=72.8 U/min/g). CONCLUSION: Four weeks after the spinal cord transection, plantaris CYTOX activity and locomotor function improved with porous tube implantation. SPONSORSHIP: Natural Sciences and Engineering Research Council. PMID- 17420774 TI - Sexuality in patients with spinal cord injuries due to attempted suicide. AB - OBJECTIVE: To evaluate the specific characteristics of sexuality in spinal cord injury (SCI) patients resulting from attempted suicide pre- and post-SCI. METHODS: From March 1998 to March 2005, n=27 were compared with a SCI group with similar characteristics. In March 2006, all patients completed a final sexual check-up. Using our database we reviewed their sexuality path. RESULTS: Suicide group (SG): Examination of personal/clinical history revealed in three how sexuality represented a possible co-factor for suicidal tendencies: one female reported a history of sexual abuse, two were homosexual (one gay and one lesbian). Five reported sexual dysfunctions. Nine females consistently showed via the Female Sexual Function Index (FSFI) at least one sexual disorder in all follow-ups; six females were in a stable relationship pre-SCI, seven during the final check-up; eight males in the SG presented erectile dysfunction using the International Index of Erectile Function (IIEF5); and eight were in a stable relationship pre-SCI and seven during the final visit. Control group (CG): Pre SCI only one male presented erectile dysfunction. Seven females consistently showed sexual disorders. 11 females and 11 males were in stable relationships pre SCI, respectively, 10 and 11 at the final check-up. In women the overall sexual satisfaction is statistically significant, correlated to the duration of the relationship P<0.05. Nine patients showed erectile dysfunction in the final follow-up. CONCLUSIONS: The SG showed a higher percentage of sexual dysfunction pre-SCI compared with CG, and they presented constant difficulty in establishing a stable relationship causing both an obstacle in the initial sexual rehabilitation and in achieving a satisfactory overall sexual life. PMID- 17420775 TI - Monoterpenoid agonists of TRPV3. AB - BACKGROUND AND PURPOSE: Transient receptor potential (TRP) V3 is a thermosensitive ion channel expressed predominantly in the skin and neural tissues. It is activated by warmth and the monoterpene camphor and has been hypothesized to be involved in skin sensitization. A selection of monoterpenoid compounds was tested for TRPV3 activation to establish a structure-function relationship. The related channel TRPM8 is activated by cool temperatures and a number of chemicals, among them the monoterpene (-)-menthol. The overlap of the receptor pharmacology between the two channels was investigated. EXPERIMENTAL APPROACH: Transfected HEK293 cells were superfused with the test substances. Evoked currents were measured in whole cell patch clamp measurements. Dose response curves for the most potent agonists were obtained in Xenopus laevis oocytes. KEY RESULTS: Six monoterpenes significantly more potent than camphor were identified: 6-tert-butyl-m-cresol, carvacrol, dihydrocarveol, thymol, carveol and (+)-borneol. Their EC(50) is up to 16 times lower than that of camphor. All of these compounds carry a ring-located hydroxyl group and neither activates TRPM8 to a major extent. CONCLUSIONS AND IMPLICATIONS: Terpenoids have long been recognized as medically and pharmacologically active compounds, although their molecular targets have only partially been identified. TRPV3 activation may be responsible for several of the described effects of terpenoids. We show here that TRPV3 is activated by a number of monoterpenes and that a secondary hydroxyl-group is a structural requirement. PMID- 17420776 TI - GW3965, a synthetic liver X receptor (LXR) agonist, reduces angiotensin II mediated pressor responses in Sprague-Dawley rats. AB - BACKGROUND AND PURPOSE: Liver X receptors (LXRs) activate genes that regulate lipid and cholesterol metabolism. LXR agonists were shown recently to also increase murine renin gene expression in vivo. To further examine a link between lipid metabolism, the renin-angiotensin-aldosterone-system and blood pressure regulation, we investigated the effect of a LXR agonist (GW3965) on angiotensin II (Ang II)-mediated vasoreactivity and vascular angiotensin II receptor (ATR) gene expression. EXPERIMENTAL APPROACH: Arterial blood pressure (BP) was measured during Ang II infusions (1.5 min duration; 0.001-3 microg kg(-1)) in pentobarbital-anesthetized male Sprague-Dawley rats (n = 6-9) after oral administration of GW3965 (10 mg kg(-1), q.d.) or vehicle for 7 - 15 days. Mesenteric arteries and plasma were collected to analyze ATR gene expression and to measure plasma renin activity (PRA) and lipid profile, respectively. KEY RESULTS: Basal mean arterial pressure (MAP) was similar between groups. GW3965 dosing blunted the vasopressor effect of Ang II, which was significantly different with the 0.3 and 3 microg kg(-1) doses. No difference in heart rate, PRA or lipid profile was observed between groups. A time-course indicated that ATR type 1 and 2 gene expression of GW3965-treated vs. vehicle-treated rats decreased by 50%, reaching significance for ATR type 2, but not for ATR type 1, at time-points coinciding with BP measurements. CONCLUSIONS AND IMPLICATIONS: GW3965 decreased Ang II-mediated vasopressor responses coincident with a trend toward reduced ATR gene expression, suggesting that LXR agonists could affect vascular reactivity. PMID- 17420777 TI - Thromboxane A2 inhibition of SKCa after NO synthase block in rat middle cerebral artery. AB - BACKGROUND AND PURPOSE: NO/prostanoid independent, EDHF-mediated hyperpolarization and dilation in rat middle cerebral arteries is mediated solely by endothelial cell IK(Ca). However, when the NO-pathway is also active, both SK(Ca) and IK(Ca) contribute to EDHF responses. As the SK(Ca) component can be inhibited by stimulation of thromboxane A(2) (TxA(2)) TP receptors and NO has the potential ability to inhibit thromboxane synthesis, we investigated whether TxA(2) might explain loss of functional input from SK(Ca) during NOS inhibition in cerebral arteries. EXPERIMENTAL APPROACH: Rat middle cerebral arteries were mounted in a wire myograph. Endothelium-dependent responses to the PAR2 agonist, SLIGRL were assessed as simultaneous changes in smooth muscle membrane potential and tension. KEY RESULTS: Responses were obtained in the presence of L-NAME as appropriate. Inhibition of TP receptors with either ICI 192,605 or SQ 29,548, did not affect EDHF mediated hyperpolarization and relaxation, but in their presence neither TRAM-34 nor apamin (to block IK(Ca) and SK(Ca) respectively) individually affected the EDHF response. However, in combination they virtually abolished it. Similar effects were obtained in the presence of the thromboxane synthase inhibitor, furegrelate, which additionally revealed an iberiotoxin-sensitive residual EDHF hyperpolarization and relaxation in the combined presence of TRAM 34 and apamin. CONCLUSIONS AND IMPLICATIONS: In the rat middle cerebral artery, inhibition of NOS leads to a loss of the SK(Ca) component of EDHF responses. Either antagonism of TP receptors or block of thromboxane synthase restores an input through SK(Ca). These data indicate that NO normally enables SK(Ca) activity in rat middle cerebral arteries. PMID- 17420778 TI - Pharmacokinetic-pharmacodynamic modelling of S(-)-atenolol in rats: reduction of isoprenaline-induced tachycardia as a continuous pharmacodynamic endpoint. AB - BACKGROUND AND PURPOSE: For development of mechanism-based pharmacokinetic pharmacodynamic (PK-PD) models, continuous recording of drug effects is essential. We therefore explored the use of isoprenaline in the continuous measurement of the cardiovascular effects of antagonists of beta-adrenoceptors (beta-blockers). The aim was to validate heart rate as a pharmacodynamic endpoint under continuous isoprenaline-induced tachycardia by means of PK-PD modelling of S(-)-atenolol. EXPERIMENTAL APPROACH: Groups of WKY rats received a 15 min i.v. infusion of 5 mg kg(-1) S(-)-atenolol, with or without i.v. infusion of 5 microg kg(-1) h(-1) isoprenaline. Heart rate was continuously monitored and blood samples were taken. KEY RESULTS: A three-compartment model best described the pharmacokinetics of S(-)-atenolol. The PK-PD relationship was described by a sigmoid Emax model and an effect compartment was used to resolve the observed hysteresis. In the group without isoprenaline, the variability in heart rate (30 b.p.m.) approximated the maximal effect (Emax=43+/-18 b.p.m.), leaving the parameter estimate of potency (EC50=28+/-27 ng ml(-1)) unreliable. Both precise and reliable parameter estimates were obtained during isoprenaline-induced tachycardia: 517+/-13 b.p.m. (E0), 168+/-15 b.p.m. (Emax), 49+/-14 ng ml(-1) (EC50), 0.042+/-0.012 min(-1) (k(eo)) and 0.95+/-0.34 (n). CONCLUSIONS AND IMPLICATIONS: Reduction of heart rate during isoprenaline-induced tachycardia is a reliable pharmacodynamic endpoint for beta-blockers in vivo in rats. Consequently this experimental approach will be used to investigate the relationship between drug characteristics and in vivo effects of different beta blockers. PMID- 17420779 TI - Differential effects of selective oestrogen receptor modulators (SERMs) tamoxifen, ospemifene and raloxifene on human osteoclasts in vitro. AB - BACKGROUND AND PURPOSE: Several selective oestrogen receptor modulators (SERMs) with oestrogen agonist effects in bone cells and without increased risk of breast and endometrial cancer have been developed. Here, we have investigated the effects of different types of SERMs on osteoclast differentiation, bone resorption and apoptosis in vitro. EXPERIMENTAL APPROACH: Human peripheral blood derived CD14+ monocytes were cultured on bovine bone slices in the presence of RANKL, M-CSF, TNF-alpha and dexamethasone for seven days. Also, CD14+ monocytes were co-cultured either with human SaOS-2 or MG-63 osteosarcoma cells, in the presence of parathyroid hormone. Osteoclast cultures were treated with different SERMs. TRACP+ multinucleated cells and C-terminal telopeptide of type I collagen were used as markers for osteoclast formation and bone resorption, respectively. KEY RESULTS: In CD14+ monocyte cultures, tamoxifen directly inhibited human osteoclast formation and bone resorption, while raloxifene and ospemifene had no inhibitory effect. In the co-cultures either with SaOS-2 or MG-63 cells, ospemifene and raloxifene as well as tamoxifen inhibited osteoclast formation in a concentration-dependent manner. The inhibitory effect was associated with an increased production of osteoprotegerin. The anti-oestrogen ICI 182 780 completely reversed the effects of these SERMs. CONCLUSION AND IMPLICATIONS: Tamoxifen had an oestrogen receptor dependent, direct, inhibitory effect on human osteoclast differentiation and bone resorption, whereas ospemifene and raloxifene required osteoblastic cells to achieve a similar inhibition. The effects of ospemifene and raloxifene were mediated by oestrogen receptors by a mechanism involving paracrine induction of osteoprotegerin in cultures with osteoblast derived osteosarcoma cells. PMID- 17420780 TI - Antileukoproteinase protects against hepatic inflammation, but not apoptosis in the response of D-galactosamine-sensitized mice to lipopolysaccharide. AB - BACKGROUND AND PURPOSE: There is major evidence for the strong bi-directional interrelation of parenchymal cell apoptosis and leukocyte accumulation and inflammation in acute liver injury. Therefore, the aim of this in vivo study was to investigate the anti-apoptotic and anti-inflammatory potential of antileukoproteinase (ALP) in a murine model of acute liver failure. EXPERIMENTAL APPROACH: C57BL/6J mice were given galactosamine (D-GalN) and E. coli lipopolysaccharide (LPS) followed by administration of saline or ALP. Besides survival rate, hepatic tissue damage and inflammatory response were analyzed by intravital fluorescence microscopy 6 hours after treatment. In addition, immunohistochemical analysis of NFkappaB-p65 and hepatocellular apoptosis, plasma levels of AST/ALT, TNF-alpha and IL-10 were determined. KEY RESULTS: Administration of D-GalN/LPS provoked hepatic damage, including marked leukocyte recruitment and microvascular perfusion failure, as well as hepatocellular apoptosis and enzyme release. NFkappaB-p65 became increasingly detectable in hepatocellular nuclei, accompanied by a rise of TNF-alpha and IL-10 plasma levels. ALP markedly reduced intrahepatic leukocyte accumulation, nuclear translocation of NFkappaB and plasma levels of TNF-alpha and IL-10. Moreover, liver enzyme levels indicated the absence of necrotic parenchymal cell death. In contrast, ALP failed to block both apoptosis and caspase-3 levels and the mortality rate of ALP-treated animals was comparable to that of saline-treated mice. CONCLUSIONS AND IMPLICATIONS: ALP could effectively prevent D-GalN/LPS associated intrahepatic inflammatory responses by inhibition of NFkappaB activity, but not apoptosis-driven mortality. Thus, a protease-inactivating approach such as application of ALP seems to be inadequate in damaged liver where apoptosis represents the predominant mode of cell death. PMID- 17420781 TI - Long-term persistence with orlistat and sibutramine in a population-based cohort. AB - OBJECTIVE: Orlistat and sibutramine are widely prescribed antiobesity agents that are approved for 2 years of continuous use. Previous 1-4-year randomized, placebo controlled trials of these drugs have reported average weight losses of <5 kg, significant adverse effects and attrition rates of up to 60%. The objective of this study was to determine the long-term persistence with orlistat and sibutramine therapy outside a clinical trial setting. DESIGN, SETTING AND PATIENTS: Population-based administrative data from British Columbia, Canada, were used to create an inception cohort of orlistat and sibutramine users and determine the 2-year persistence with therapy. MAIN OUTCOME MEASURE: Persistence with therapy at 2 years. Drug discontinuation was defined as the failure to refill a prescription within 120 days. Patients discontinuing therapy were censored at the 60-day mark. RESULTS: Nearly 17 000 users of orlistat and 3500 users of sibutramine were identified. For both orlistat and sibutramine, 1-year persistence rates were <10% and 2-year persistence rates were 2%. CONCLUSION: This population-based, retrospective cohort analysis demonstrated very poor long term persistence rates with orlistat and sibutramine and discontinuation rates that were much higher than those reported in clinical trials. PMID- 17420782 TI - Could a virus contribute to weight gain? AB - OBJECTIVE: Obesity is a serious public health problem associated with increased morbidity and mortality. Although the causes for obesity are unclear, it seems that environmental, genetic, neural and endocrine factors contribute to its development. However, the rapid global spread of obesity resembles epidemiologically the spread of an infectious disease. Thus far, little consideration has been given to the possibility that the epidemic of obesity could be due to an infectious agent. Seven viruses and a scrapie agent have been implicated in obesity. DESIGN: This review evaluates the infectious pathogens and the evidence that these viruses are associated with obesity and concludes that a strong evidence base is emerging that associates certain viruses with obesity. CONCLUSION: More work is however required to elucidate the mechanisms of weight gain after viral infection. In the mean time, discounting viruses as a contributing factor to obesity would deprive us of a potential new avenue of investigating and treating the ever increasing epidemic of obesity. PMID- 17420783 TI - The "father of modern interventional pediatric cardiology" retires. PMID- 17420784 TI - These are the days: the internship revisited. PMID- 17420785 TI - Cardiovascular disease in Africa. PMID- 17420786 TI - Open-heart surgery in Nigeria: indications and challenges. PMID- 17420787 TI - Systemic infections cause exaggerated local inflammation in atherosclerotic coronary arteries: clues to the triggering effect of acute infections on acute coronary syndromes. AB - Systemic infections can trigger heart attacks. We conducted an autopsy study to investigate the pathologic effect of systemic infections on coronary artery inflammation. We studied 14 atherosclerotic patients diagnosed with an acute systemic infection. Our control group (n=13) had atherosclerosis without infection. The groups were similar in luminal stenosis and age. Coronary artery sections were stained with H&E and markers for macrophages (CD68), T cells (CD3), and dendritic cells (S100). On pathologic examination, 5 infected patients had acute myocardial infarction with thrombosis. Macrophage density in plaques and in periadventitial fat was higher in the infected group (NS). The infected patients' adventitia had significantly more macrophages (1,577 +/- 1,872 vs 265 +/- 185 per mm(2); P=0.047). The macrophage density, similar in the control group's adventitia and plaque, was significantly greater in the infected group's adventitia than in the plaque. The adventitia and periadventitial fat of the infected group had more T cells than did samples from the control group (48.4 +/- 45.0 vs 14.1 +/- 6.3 per mm(2); P=0.002). The groups exhibited similar plaque T cell density. The infected patients' plaques, but not the adventitia and periadventitial fat, had more dendritic cells than did the controls' (3.2 +/- 2.5 vs 0.3 +/- 0.5 per mm(2); P=0.022). To our knowledge, this is the 1st report to establish a connection between acute systemic infections and significant increases in inflammatory cells in the atherosclerotic coronary arteries of human beings. This offers a new therapeutic target for preventing heart attacks in high risk patients. PMID- 17420788 TI - Management of air embolism during HeartMate XVE exchange. AB - Air embolism is a rare and usually fatal complication of major cardiac surgery. We present a case in which a 45-year-old man supported by a HeartMate(R) XVE left ventricular assist device required a pump exchange due to failure of the device motor. During pump dissection, a massive amount of air entered the systemic circulation. Urgent cannulation for cardiopulmonary bypass was performed, and cardiopulmonary bypass was initiated, followed by profound hypothermia, circulatory arrest, retrograde cerebral perfusion, retrograde coronary sinus perfusion, and then barbiturate coma and steroid therapy. The HeartMate XVE left ventricular assist device was removed, and a HeartMate II was implanted. After 5 days, the patient awoke with left hemiparesis, which nearly resolved with aggressive physical therapy. Forty-four days after the pump exchange operation, the patient was discharged from the hospital with only mild left hemiparesis. Exposure of the left ventricular assist device or its external components requires careful monitoring, because air can enter the pump-particularly in a hypovolemic patient. Rapid response after massive air entry into the left ventricular assist device system, as in our patient, can result in a successful outcome. PMID- 17420789 TI - Body perfusion in surgery of the aortic arch. AB - We propose a new cannulation and perfusion technique for aortic arch surgery, in order to achieve continuous antegrade total-body perfusion under moderate hypothermia. The heart and the aortic arch are exposed through a median sternotomy. Cardiopulmonary bypass is established from the right atrium to the right axillary artery. At 26 degrees C of body temperature, the supra-aortic vessels are clamped, the ascending aorta and the aortic arch are incised, and a cuffed endotracheal cannula, connected to an arterial line geared by a separate roller pump, is inserted into the descending thoracic aorta. Perfusion is started in the distal body, while the brain is perfused through the right axillary artery. Once the aortic arch has been replaced with a Dacron graft and the supra aortic vessels have been reimplanted on the graft, the arterial line in the descending thoracic aorta is clamped and removed. The supra-aortic vessel clamps are removed, the proximal part of the Dacron graft is clamped, and systemic cardiopulmonary bypass is resumed via the right axillary artery.From January 2002 through December 2005, this technique was used in 12 consecutive patients on an emergency basis, due to acute aortic dissection that required total arch replacement. Within the first 30 postoperative days, 1 patient (8.3%) died, and no patient had permanent neurologic deficits. This simple technique ensures a full-flow antegrade total-body perfusion during all phases of the surgical procedure, thereby eliminating ischemia-reperfusion syndrome and yielding excellent clinical results. PMID- 17420790 TI - Modified inferior vena caval anastomosis to reduce tricuspid valve regurgitation after heart transplantation. AB - Postoperative tricuspid valve regurgitation is moderate to severe in 15% to 20% of heart transplant recipients despite use of the bicaval surgical technique. We hypothesized that the regurgitation might be partly due to increased tension on the donor right atrium. To study the right atrial distortion, we modified the standard bicaval anastomosis. Our technique involves augmenting the donor right atrial anterior wall with a flap of the recipient's right atrium, which is left attached in continuity with the anterior aspect of the inferior vena cava along 65% of its circumference. We measured tricuspid regurgitation, right atrial area, and right atrioventricular diameter in 7 consecutive patients who underwent orthotopic heart transplantation with the modified anastomosis. Tricuspid regurgitation was graded as follows: 1 = trace, <10%; 2 = mild, 10%-24%; 3 = moderate, 25%-50%; and 4 = severe, >50%. All patients were weaned from inotropic support within 1 week after transplantation with excellent ventricular function, no heart block, and 100% survival at 30 days. The median follow-up time was 173 days (44-358 days). Other median measurements included tricuspid valve regurgitation jet area, 0.30 cm(2) (0-1.90 cm(2)); right atrial area, 15.90 cm(2) (14.47-18.00 cm(2)); atrioventricular diameter, 2.70 cm (2.63-3.09 cm); and tricuspid regurgitation, 1.67% (0-12.42%). Mild regurgitation occurred in 1 recipient; in all others, it was trace. The modified inferior vena caval anastomosis is simple and safe. It eliminates moderate and severe tricuspid valve regurgitation without routine annuloplasty after orthotopic heart transplantation via the bicaval technique. PMID- 17420791 TI - Percutaneous ventricular assist during aortic valvuloplasty: potential application to the deployment of aortic stent-valves. AB - We evaluated the short-term safety and efficacy of using the TandemHeart(R) percutaneous ventricular assist device in high-risk patients undergoing aortic valvuloplasty procedures. Aortic valvuloplasty was performed in 4 patients who had no ventricular assist device support and in 7 patients who used the TandemHeart for hemodynamic support. The age range was 65 to 94 years (mean, 83 +/- 11 yr). The mean ejection fraction was 0.30 +/- 0.14. A transseptal antegrade approach to the aortic valve was used in 8 patients and a retrograde approach in the remaining 3. WITH THE TANDEMHEART, ALL PROCEDURES WERE TECHNICALLY SUCCESSFUL: each patient survived at least 1 month after the procedure. The mean total balloon inflation time was 37 +/- 10 sec. The aortic valve area was 0.6 +/- 0.1 cm(2) before the procedure and 0.9 +/- 0.2 cm(2) afterwards (P=0.006). Without TandemHeart support, 1 patient died of cardiac arrest during the procedure. The mean total balloon inflation time was 11 +/- 3 sec. Aortic valve area was 0.6 +/- 0 cm(2) before the procedure and 1.1 +/- 0.3 cm(2) afterwards (P=0.3). No patient developed aortic regurgitation. We conclude that use of the TandemHeart for hemodynamic support during high-risk aortic valvuloplasty is associated with favorable intraprocedural and short-term outcomes. With the TandemHeart in place, balloon placement was precise, and inflation was maintained for up to 45 sec without balloon displacement. These attributes are essential during stent-valve placement, are achieved without rapid ventricular pacing, and may reduce the risk of global ischemia and death. PMID- 17420792 TI - Acute massive pulmonary embolism with cardiopulmonary resuscitation: management and results. AB - Patients who experience hemodynamic collapse after acute massive pulmonary embolism have a poor prognosis. Herein, we report our results with 8 patients and discuss a surgical strategy that can improve perioperative survival. From August 1994 through May 2005, 8 consecutive patients (6 women, 2 men; age range, 27-68 yr) were urgently referred to our unit after experiencing hemodynamic collapse. All required cardiopulmonary resuscitation. Seven patients underwent pulmonary embolectomy. One patient was successfully treated with thrombolytic therapy alone under continuous monitoring by the surgical team. There were 2 intraoperative deaths (30-day mortality rate, 28.5%). One survivor required a right ventricular assist device. Follow-up of the patients ranged from 8 months to 8 years. One patient died 8 months after the pulmonary embolectomy from long-term complications of cerebral damage that had occurred during preoperative resuscitation. We conclude that prompt surgical management improves the early survival rates of patients who require cardiopulmonary resuscitation subsequent to massive pulmonary embolism. PMID- 17420793 TI - Surgical treatment of post-infarction left ventricular pseudoaneurysm: a two decade experience. AB - Herein, we present a retrospective analysis of our experience with acquired pseudoaneurysms of the left ventricle over a 20-year period.From February 1985 through September 2004, 14 patients underwent operation for left ventricular pseudoaneurysm in our clinic. All pseudoaneurysms (12 chronic, 2 acute) were caused by myocardial infarction. The mean interval between myocardial infarction and diagnosis of pseudoaneurysm was 7 months (range, 1-11 mo). The pseudoaneurysm was located in the inferior or posterolateral wall in 11 of 14 patients (78.6%). In all patients, the pseudoaneurysm was resected and the ventricular wall defect was closed with direct suture (6 patients) or a patch (8 patients). Most patients had 3-vessel coronary artery disease. Coronary artery bypass grafting was performed in all patients. Five patients died (postoperative mortality rate, 35.7%) after repair of a pseudoaneurysm (post-infarction, 2 patients; chronic, 3 patients). Two patients died during follow-up (median, 42 mo), due to cancer in 1 patient and sudden death in the other. Although repair of left ventricular pseudoaneurysm is still a surgical challenge, it can be performed with acceptable results in most patients. Surgical repair is warranted particularly in cases of large or expanding pseudoaneurysms because of the propensity for fatal rupture. PMID- 17420794 TI - Effects of carvedilol on plasma levels of pro-inflammatory cytokines in patients with ischemic and nonischemic dilated cardiomyopathy. AB - We prospectively investigated the effects of adding carvedilol to the standard treatment of ischemic and nonischemic dilated cardiomyopathy (DCM), by measuring the plasma levels of pro-inflammatory cytokines. Sixty patients with DCM (35 ischemic and 25 nonischemic) were divided into 2 subgroups: patients on standard therapy alone (digoxin, angiotensin-converting enzyme inhibitors, and diuretics) and patients on standard therapy plus carvedilol. Study participants' serum levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2), and interleukin-6 (IL-6) were measured at the beginning and again at the end of the study. Left ventricular ejection fraction and left ventricular diastolic function were evaluated by means of radionuclide ventriculography. In ischemic patients on carvedilol, levels of IL-6 and TNF-alpha dropped significantly (P= 0.028 and P=0.034, respectively). In ischemic patients on standard treatment, plasma IL-2 levels were elevated after treatment (P=0.047). No significant differences occurred in IL-6 levels, while TNF-alpha levels were elevated (P=0.008). In nonischemic patients on carvedilol, IL-6 and TNF-alpha levels dropped significantly (P=0.018 and P=0.004, respectively). The left ventricular ejection fraction increased significantly (P=0.006). In nonischemic patients on standard treatment, no significant change occurred in any value. Carvedilol suppressed the plasma levels of TNF-alpha and IL-6 in both ischemic and nonischemic patients. The carvedilol effect was more pronounced in patients with nonischemic dilated cardiomyopathy than in those with ischemic disease. PMID- 17420795 TI - Iliac vein stenting for chronic venous insufficiency. AB - Chronic venous insufficiency has devastating sequelae in terms of patients' lifestyles and negative economic impact on society. Traditional surgical procedures have yielded variable patency results, and follow-up has not always been reported. This review summarizes the current applications, patency rates, stent selection, and complications of balloon angioplasty and stenting in the treatment of chronic venous outflow obstruction in the lower extremity. We conclude that endovenous stenting is the current method of choice in the treatment of chronic venous obstruction. PMID- 17420796 TI - Inherited arrhythmic disorders: long QT and Brugada syndromes. AB - Inherited arrhythmic disorders comprise a group of syndromes with unique genetic abnormalities and presentations but with very similar clinical outcomes and complications, the most terrifying of which are life-threatening arrhythmias and sudden cardiac death. Advances in molecular biology have enabled us to define and pinpoint many such disorders, which were previously labeled as idiopathic, to specific genes on various chromosomes. The current trend in the management of these potentially deadly disorders is to use pharmacotherapy (antiarrhythmic agents) and defibrillators for the prevention of sudden death; however, targeted therapy at a molecular level appears to be the path of the future. Herein, we review long QT and Brugada syndromes and focus on the genetics, pathophysiology, and clinical manifestations of these inherited arrhythmogenic disorders that affect patients with structurally normal hearts. PMID- 17420797 TI - Takotsubo cardiomyopathy, or broken-heart syndrome. AB - Takotsubo cardiomyopathy mimics acute coronary syndrome and is accompanied by reversible left ventricular apical ballooning in the absence of angiographically significant coronary artery stenosis. In Japanese, "takotsubo" means "fishing pot for trapping octopus," and the left ventricle of a patient diagnosed with this condition resembles that shape. Takotsubo cardiomyopathy, which is transient and typically precipitated by acute emotional stress, is also known as "stress cardiomyopathy" or "broken-heart syndrome."Herein, we describe the clinical angiographic characteristics of 4 patients who exhibited this syndrome, and we review the existing literature and propose reasons to conduct prospective studies. PMID- 17420798 TI - Intravenous adenosine for surgical management of penetrating heart wounds. AB - Accurate suturing of penetrating cardiac injuries is difficult. Heart motion, ongoing blood loss, arrhythmias due to heart manipulation, and the near-death condition of the patient can all affect the outcome. Rapid intravenous injection of adenosine induces temporary asystole that enables placement of sutures in a motionless surgical field. Use of this technique improves surgical conditions, and it is faster than other methods. Herein, we describe our experience with the use of intravenous adenosine to successfully treat 3 patients who had penetrating heart wounds. PMID- 17420799 TI - Isolated chylopericardium after intrapericardial procedures: possible role of inadvertent right efferent lymphatic trunk injury. AB - Chylopericardium after an intrapericardial procedure is rare, and satisfactory explanations of its possible causes are lacking.Herein, we present 4 cases of chylopericardium that developed after intrapericardial surgery, and we review the literature. Our literature review revealed 29 cases of chylopericardium that complicated intrapericardial operations, to which we added our 4 cases for analysis. The 33 surgical procedures involved repair for congenital heart disease (n=21), valve surgery (n=5), coronary artery bypass grafting (n=6), and other (n=1). Causes were verified in 7 patients: small lymphatic injury in 3 and high venous pressure or venous thrombosis in 4. Of the 26 patients with chylopericardium of unknown origin, 15 had congenital heart disease. Ten of these 15 had chromosomal abnormalities, especially trisomy 21 (Down syndrome); these patients typically had increased lymphatic permeability, which raised the likelihood of chylopericardium. Five revascularizations for coronary artery disease required harvesting of the left internal thoracic artery for reconstruction, incurring a risk of damage to the drainage site of the right efferent lymphatic trunk. In addition, all 26 patients with chylopericardium of unknown origin underwent dissection of the ascending aorta and the main pulmonary artery, near the right efferent lymphatic trunk. Inadvertent injury to the trunk during the dissection would have increased the risk of chylopericardium. Accordingly, even though the overall incidence of chylopericardium during intrapericardial procedures is low, we recommend a meticulous dissection of the ascending aorta from the main pulmonary artery. PMID- 17420800 TI - Pericardial effusion as an expression of thyrotoxicosis. AB - Patients who have either hyperthyroidism or hypothyroidism can present with cardiovascular complications. These manifestations of thyroid disease-congestive heart failure, atrial tachyarrhythmias, atrioventricular conduction disorders, and mitral valve dysfunction-are well known to the clinician. Pericardial effusion is considered a complication of hypothyroidism; as an expression of thyrotoxicosis, it is extremely rare.Herein, we present the case of a 76-year-old woman who had pericardial effusion associated with thyrotoxicosis. She was treated with high-dose beta-blockers, methimazole, diuretics, and short-term steroids. She recovered completely, which precluded the need for pericardiocentesis. We suggest that thyrotoxicosis be considered in the differential diagnosis of pericardial effusion. PMID- 17420801 TI - Infective coronary aneurysms: a complication of percutaneous coronary intervention. AB - We present the case of a patient who developed infective coronary artery aneurysms after percutaneous coronary artery intervention. We describe the patient's clinical presentation, diagnosis, and treatment, and we review the pertinent medical literature. PMID- 17420802 TI - Aspergillus endocarditis of the mitral valve in a lung-transplant patient. AB - A 57-year-old man underwent bilateral lung transplantation at our hospital. On histopathology, aspergillomas were identified in the upper lobes of the explanted lungs. After being treated and discharged from the hospital, he returned 4 months later with ischemic chest pain, which was due to a myocardial infarction complicated by cardiogenic shock. He also had a large vegetation on the anterior mitral leaflet. Herein, we describe the patient's symptoms, complications, treatment, and recovery. To the best of our knowledge, ours is only the 2nd report of a patient who developed Aspergillus endocarditis after lung transplantation and the 1st such patient to have undergone successful mitral valve replacement. PMID- 17420803 TI - Transvenous right atrial and left ventricular pacing after the Fontan operation: long-term hemodynamic and electrophysiologic benefit of early atrioventricular resynchronization. AB - We report a case of long-term, successful, endocardial atrioventricular pacing in a 32-year-old man who had severe heart failure and ascites after having undergone a Fontan procedure for tricuspid atresia 9 years earlier. The patient was referred to our hospital for Fontan revision. However, electroanatomic mapping of the right atrium revealed viable tissue at the interatrial septum above the os of the coronary sinus, and it appeared that the left ventricle could be paced from a coronary sinus branch. Therefore, instead of Fontan revision, an endocardial atrioventricular pacemaker was implanted transvenously. On 5-year follow-up, the patient remained in New York Heart Association functional class I and had not been readmitted to the hospital for congestive heart failure or arrhythmias. His atrial and ventricular leads continued to show excellent pacing and sensing results. PMID- 17420804 TI - C-ring mitral annuloplasty: 27-year follow-up. AB - We report the long-term follow-up of a patient with rheumatic mitral valve disease who underwent annuloplasty with a specially developed C-ring (the Cooley C-ring) for mitral valve repair in 1977. The repaired valve remained competent and unobstructed for 27 years before requiring replacement. PMID- 17420805 TI - Color Doppler imaging of the ophthalmic artery during antegrade selective cerebral perfusion. AB - Postoperative brain damage is a major sequela of cardiovascular surgery. Different methods-such as transcranial Doppler imaging, carotid echography, and fluorescein angiography-can be used to monitor cerebral blood flow, but they present specific limitations.High-resolution color Doppler imaging of the ophthalmic artery is a useful, noninvasive, and inexpensive technique for the evaluation of cerebral blood flow during cardiac surgery; signal changes can represent hypoperfusion of cerebral vessels and can thereby indicate inadequate cerebral perfusion during cardiac surgery, especially surgery of the aortic arch. PMID- 17420806 TI - Transfusion-free cardiac reoperation in an 11-kg Jehovah's Witness child by use of a minimized cardiopulmonary bypass circuit. AB - Herein, we describe the design of a perfusion system for a complex cardiovascular reoperation in an 11-kg Jehovah's Witness patient. The goal of safe, transfusion free surgery was achieved chiefly by minimizing the priming volume of the cardiopulmonary bypass circuit to 200 mL while providing adequate flow and standard safety features. PMID- 17420807 TI - Gastric volvulus after ventricular assist device explantation and cardiac transplantation. AB - Diaphragmatic herniation has been recognized as a complication of unrepaired diaphragmatic defects after left ventricular assist device explantation and cardiac transplantation. We believe this to be the 1st report of diaphragmatic herniation that resulted in gastric volvulus in a cardiac transplant recipient. The presentation, diagnosis, and treatment of this potentially fatal condition are discussed herein. Nine months after removal of a Thoratec HeartMate II left ventricular assist device and orthotopic cardiac transplantation, the patient presented with intermittent upper abdominal and lower chest discomfort of 3 weeks' duration. Physical examination was notable for fullness in the upper abdomen. Plain radiographs and computed tomographic scans of the chest and abdomen without contrast were unexceptional. Two weeks later, the patient's pain began to worsen rapidly, and an upper gastrointestinal barium study revealed partial herniation of the stomach into the chest and omento-axial gastric volvulus without luminal obstruction. The patient underwent uncomplicated laparotomy for repair of the diaphragmatic defect and reduction of the herniated stomach. This case highlights the need for increased awareness of diaphragmatic herniation as a complication of unrepaired diaphragmatic defects so that diagnosis is not delayed, and underscores the importance of primary repair of all such defects to prevent future complications. PMID- 17420808 TI - Bivalirudin anticoagulation for cardiopulmonary bypass: an unusual case. AB - The standard agent used for systemic anticoagulation during cardiopulmonary bypass is heparin. Alternative methods of anticoagulation are required for patients with heparin hypersensitivity. We present the case of a patient with heparin hypersensitivity who was anticoagulated with bivalirudin during cardiopulmonary bypass for coronary artery bypass grafting. This presented unusual challenges surrounding the monitoring of anticoagulation and the method of myocardial protection. PMID- 17420809 TI - Intimal sarcoma of the pulmonary artery with retrograde extension into the pulmonic valve and right ventricle. AB - We describe the case of a 42-year-old man who presented with dyspnea on exertion and a history of anticoagulation therapy for what was thought to be pulmonary arterial thromboembolism. He underwent surgery for obstruction of the right ventricular outflow tract. This is a very rare case of an intimal sarcoma of the pulmonary artery, which we confirmed by pathologic studies. PMID- 17420810 TI - Left atrial myxosarcoma with previously detected intestinal metastasis. AB - Primary cardiac myxosarcoma is a rare disease; it is exceedingly rare for symptoms of systemic metastasis to precede diagnosis of the primary cardiac tumor. We describe the case of a previously healthy 60-year-old man with left atrial myxosarcoma, who had first presented with jejunal intussusception due to intestinal polyposis. Three months after resection of the jejunum, the patient experienced cerebral infarction and pulmonary edema. Further physical evaluation, which included echocardiography for the 1st time, revealed a mass in the left atrium that protruded through the mitral valve into the left ventricle. At emergency cardiac surgery, we found that the tumor involved multiple sites of the left atrium, the pulmonary veins, and the mitral anterior leaflet. Two months after surgery, the patient died of massive cerebral hemorrhage. Necropsy disclosed multiple recurrences of the cardiac myxosarcoma and widespread metastatic lesions. The intestinal polyps that had been resected originally were diagnosed, on retrospective histopathologic examination, as metastases of the myxosarcoma. In this unusual case, the metastatic lesions were the 1st clinical manifestations of a malignant cardiac tumor. PMID- 17420811 TI - Left atrial hemangioma presenting as cardiac tamponade. PMID- 17420812 TI - Large left ventricular capillary hemangioma with cavernous areas. PMID- 17420813 TI - Inferolateral left ventricular aneurysm preventing mitral regurgitation. PMID- 17420814 TI - Metastatic cardiac carcinoid. PMID- 17420815 TI - Acute spinal cord ischemia during aortography. PMID- 17420816 TI - Acute myocardial infarction with normal coronary arteries in a patient with Hodgkin's disease. PMID- 17420817 TI - First, do no harm: the risks of overtreating children with epilepsy. AB - BACKGROUND: Although overtreatment with antiepileptic drugs contributes to the morbidity associated with epilepsy, many children still are overtreated. OBJECTIVE: To evaluate if the withdrawal of at least one antiepileptic drug (AED) in children with refractory epilepsy using polytherapy enable a better seizure control. METHOD: This was a prospective study. Children with refractory epilepsy using at least two AEDs were included. Once the patient, or guardian, agreed to participate in the study, one or more AED were slowly tapered off. The remaining AEDs dosages could be adjusted as needed, but a new AED could not be introduced. RESULTS: Fifteen patients were evaluated, three girls; ages ranging from 3 to 18 (mean=8.7 years). After at least one AED withdrawal, two (13.5%) patients became seizure free, seizures improved >50% in 5 (33.5%) patients, did not change in 5 (33.5%), and seizure frequency became worse in 3 (20%). Adverse events improved in 12 patients (80%). CONCLUSION: The withdrawal of at least one AED is a valuable option in the treatment of selected children with refractory epilepsy. PMID- 17420818 TI - Early-onset Parkinson's disease and depression. AB - Patients with Parkinsons disease (PD) in whom symptoms start before the age of 45 years (EOPD) present different clinical characteristics from those with the late onset form of the disease. The incidence of depression is believed to be greater in patients with EOPD than with the late-onset form of the disease, although there is no risk factor or marker for depression in patients with PD. We studied 45 patients with EOPD to define the frequency of depression and to identify possible differences between the groups with and without depression. Depression was diagnosed in 16 (35.5%) of the patients, a higher incidence than in the population at large but similar to the figure for late-onset Parkinson disease; 8 (50%) of the patients had mild depression, 4 (25%) moderate depression and 4 (25%) were in remission. There was no relationship between depression and any of the clinical characteristics of the disease, although the EOPD patients with depression presented earlier levodopa-related complications and were more affected on the Hoehn-Yahr, UPDRS and Schwab-England scales. PMID- 17420819 TI - Association analysis between the C516T polymorphism in the 5-HT2A receptor gene and schizophrenia. AB - Data from epidemiological studies have demonstrated that genetics is an important risk factor for schizophrenia. Disturbances of serotonergic brain pathways have been implicated in the pathophysiology of schizophrenia. Some studies have suggested that the efficacy of atypical antipsychotics on schizophrenia treatment may be related to the serotonin 2A receptor (5-HT2A), and that serotonergic drugs may induce psychotic symptoms. Thus, the aim of this study was to investigate the association between the C516T polymorphism and schizophrenia in a Brazilian population composed by 246 patients and 315 healthy matched controls in a case control approach. No statistically differences were observed in allelic (chi2=1.77, 1d.f., p=0.18) or genotypic (chi2=1.69, 2d.f., p=0.42) distributions between cases and controls. The results suggest that the C516T polymorphism of the 5-HT2A receptor gene is not related to the susceptibility for schizophrenia in our Brazilian sample. PMID- 17420820 TI - Study of polymorphisms in the interleukin-4 and IL-4 receptor genes in a population of Brazilian patients with multiple sclerosis. AB - This study aimed to investigate in a population of Brazilian patients with multiple sclerosis (MS) single-nucleotide polymorphisms (SNP) in the promoter region of IL4 (*33C-T) and receptor IL4R (*Q551R A-G) genes proposed to interfere with disease progression. No significant differences were observed in either of the SNPs investigated between healthy controls (n=135) and MS patients (n=129). However, the IL4+33 TT genotype was significantly (p=0.039) higher in African descendants MS (AF-MS= 9.09%) than in Caucasian MS (CA-MS= 1.35%). It was also observed a significant (p=0.016) increase for the IL4R* Q551R CC genotype in AF MS compared to those of Caucasian ethnicity (AF-MS= 21.62%; CA-MS= 4.35%). These results suggest that IL4+33 and IL4R*Q551 polymorphisms may have a disease promoting role of TH2 mediators in African MS descendants. Additionally neither IL4 nor IL4R genes are susceptibility factors for Brazilian MS but may be able to modify ethnicity-dependent disease risk and penetrance of susceptibility factors. PMID- 17420821 TI - Linkage study of voltage-gated potassium channels in familial mesial temporal lobe epilepsy. AB - Voltage-gated potassium channels (VGKCs) play a critical role in the regulation of neuronal excitability and have been implicated in some types of epilepsies. Recently, autoimmune limbic encephalitis (LE) was associated with antibodies against VGKC. In addition, patients with LE showed partial epilepsy and increased T2 signal abnormalities in limbic structures. We have reported familial mesial temporal lobe epilepsy (FMTLE) associated with hippocampal atrophy (HA) and other signs of mesial temporal sclerosis detected by magnetic resonance imaging (MRI). In order to investigate whether VGKC may be associated to HA present in FMTLE, we perform linkage study in these candidate genes. Seventy-three microsatellites markers were genotyped in different human autosomal chromosome. Two-point LOD scores did not show evidence for linkage with any of the microsatellite markers genotyped (Zmax ranging from 0.11to-9.53 at theta=0.00). In the present study, linkage data showed no evidence that VGKC are involved in the determination of HA in FMTLE. PMID- 17420822 TI - Modelling of increased homocysteine in ischaemic stroke: post-hoc cross-sectional matched case-control analysis in young patients. AB - BACKGROUND & PURPOSE: Hyperhomocysteinaemia has been postulated to participate in pathogenesis of ischaemic stroke (IS). However, especially in young adults, there is possibility of significantly increased IS risk due to increased normal homocysteinaemia, i.e., hidden (pathologically dormant) prevalence within a healthy, normally-defined range. We performed a post-hoc modelling investigation on plasma total homocysteinaemia (THCY) in gender- and age-matched young patients in the acute IS phase. We evaluated relationships between THCY and prevalence of other potential risk factors in 41 patients vs. 41 healthy controls. METHOD: We used clinical methods, instrumental and neuroimmaging procedures, risk factors examination, total plasma homocysteine measurements and other laboratory and statistical modelling techniques. RESULTS: IS patients and healthy controls were similar not only for matching variables, but also for smoking, main vitamin status, serum creatinine and lipid profile. Patients with IS, however, had lower vitamin B6 levels and higher THCY, fibrinogen and triglycerides (TGL). At multivariate stepwise logistic regression only increased THCY and TGL were significantly and independently associated with the risk for stroke (72% model accuracy, p model=0.001). An increase of THCY with 1.0 micromol/L was associated with 22% higher risk of ischaemic stroke [adjusted OR=1.22 (95%CI 1.03?1.44)]. In this way, novel lower cut-off value for HCY of 11.58 micromol/L in younger patients has been revealed (ROC AUC= 0.67, 95CI% 0.55-0.78, p=0.009). CONCLUSION: The new THCY cut-off clearly discriminated between absence and presence of IS (sensitivity>63%, specificity>68%) irrespectively of age and gender and may be applied to better evaluate and more precisely define, as earlier as possible, the young patients at increased IS risk. PMID- 17420823 TI - A Brazilian family with Brown-Vialetto-van Laere syndrome with autosomal recessive inheritance. AB - We report the first Brazilian family with Brown-Vialetto-van Laere syndrome. The presence of consanguineous marriages and illness affecting three sisters and one niece support an autosomal recessive transmission. The age at onset of the illness ranged from 12 to 20 years old. The time interval between hearing loss and involvement of other cranial nerves varied from 3 to 12 years. MRI demonstrated bulbar atrophy and also high intensity signal at T2 weighted and fluid attenuated inversion recovery (FLAIR) sequences. PMID- 17420824 TI - Rett syndrome: clinical and molecular characterization of two Brazilian patients. AB - BACKGROUND: Rett syndrome (RS) is recognized as a pan-ethnic condition. Since the identification of mutations in the MECP2 gene, more patients have been diagnosed, and a broad spectrum of phenotypes has been reported. There is a lack of phenotype-genotype studies. OBJECTIVE: To describe two cases of Brazilian patients with identified MECP2 mutations. METHOD: We present two female Brazilian patients with RS. RESULTS: Both patients presented with regression at 2-3 years of age, when stereotypic hand movements, social withdrawal and postnatal deceleration of head growth rate were observed. Both patients presented verbal communication impairment. Case 1 had loss of purposeful hand movements, and severe seizure episodes. Case 2 had milder impairment of purposeful hand movements, and no seizures. They had different mutations, D97Y and R294X, found in exons 3 and 4 of MECP2 gene, respectively. CONCLUSION: Testing for MECP2 mutations is important to confirm diagnosis and to establish genotype/phenotype correlations, and improve genetic counseling. PMID- 17420825 TI - Cerebrovascular disease in childhood: neuropsychological investigation of 14 cases. AB - There are few studies about the development of a child after a cerebrovascular accident (CVA), and they usually describe problems such as diminishing in intellectual capacities, difficulties in linguistic and visual-motor skills, as well as in spatial organization and integration. In this study, there were 28 children participating, being 14 placed in the experimental group (EG) after clinical diagnosis and ischemic CVA imaging, and other 14 children without past history of CVA, who formed the control group (CG). The neuropsychological research protocol included an intelligence test, a visual-motor coordination test, human figure drawing, a cortical functions battery and the medical records of the children from the EG. The analysis of the results of this study revealed that the best performances after the CVA are related to the shortest time of functional recovery; CG presented better performances than EG in all the instruments used, in cognitive, perceptual and motor skills. It has been noticed that CVA may lead to intellectual reduction in case of a recurrence of the vascular insult. PMID- 17420826 TI - Factors associated to the positive cerebrospinal fluid culture in the tuberculous meningitis. AB - Central nervous system involvement is the most common neurological complication in the course of tuberculosis. The lack of rapid and sensitive tests delays the early diagnosis. Here, we retrospectively reviewed the cerebrospinal fluid (CSF) examination of 30 patients with tuberculous meningitis confirmed by bacteriological tests (culture and/or polymerase chain reaction). The purpose of the present study was to determine the CSF parameters associated to the positive CSF culture for Mycobacterium tuberculosis in tuberculous meningitis. We found higher frequency of positive CSF culture in patients infected with HIV as well in patients with high number of neutrophils and high protein content (characteristic in the early or acute-stage patients), which suggests that the positive culture found in these patients may be associated with the presence of high bacillary load in CSF occurring in these stages. PMID- 17420827 TI - Outcome of sleepiness and fatigue scores in obstructive sleep apnea syndrome patients with and without restless legs syndrome after nasal CPAP. AB - BACKGROUND & PURPOSE: The association of obstructive sleep apnea syndrome (OSAS) and restless legs syndrome (RLS) has been reported in the literature for many years. Both conditions may be responsible for fatigue and somnolence complaints secondary to nocturnal sleep disruption. The primary concern of this study is to evaluate the outcome of fatigue and daytime sleepiness symptoms at baseline and after continuous positive air pressure (CPAP) treatment in OSAS patients with and without RLS. METHOD: A prospective and comparative study between a group of 13 patients with OSAS and a group of 17 patients with OSAS+RLS. Laboratory blood tests and polysomnography were performed at baseline. The Epworth Sleepiness Scale (ESS) and the Pichots questionnaire of fatigue/depression (PIC) were applied before and after 3 months of CPAP treatment. Results were compared. RESULTS: No significant differences were found on PSG and laboratory results at baseline. Both groups had similar ESS and PIC scores at baseline (p=0.73 and 0.08, respectively). After n-CPAP, OSAS+RLS patients showed higher ESS and PIC scores (p=0.017 and 0.03, respectively). CONCLUSIONS: Despite a favorable general response, n-CPAP seemed less effective in treating fatigue and sleepiness in the OSAS+RLS group. PMID- 17420828 TI - Event-related potentials (P300) and neuropsychological assessment in boys exhibiting Duchenne muscular dystrophy. AB - OBJECTIVE: To examine auditory cognitive evoked potentials (P300 potentials) and neuropsychological dysfunction in patients with Duchenne muscular dystrophy (DMD). METHOD: P300 potentials and neuropsychological test results were obtained from 16 healthy control boys and 20 DMD patients. Full Intelligence Quotients (IQ) were estimated for patients and control group. Mean age was 9.5 years in the DMD patient group, and 10 years in the control group (p>0.05). RESULTS: The mean IQ values were 64.35 in the DMD patients and 82.68 in the control group (p=0.01). Mean P300 values were 347.6 in the DMD group and 337.4 in the control group (p=0.14). There was no significant correlation between parameters in each group. CONCLUSION: DMD patients showed a poor performance as evaluated by P300 potential compared to the control group, although the difference was not statistically significant. Systematic alterations in neuropsychological test results were found, the differences paralleling those detected in IQ. PMID- 17420829 TI - EEG spectral coherence inter- and intrahemispheric during catching object fall task. AB - The aim of the present study was to evaluate coherence measures at Theta through qEEG during the accomplishment of a specific motor task. The sample consisted of 23 healthy individuals, both sexes, with ages varying between 25 and 40 years old. All subjects were submitted to a specific motor task of catching sequences of falling balls. A three-way ANOVA was employed for the statistical analysis, which demonstrated main effects for the following factors: time, block and position. However, there was no interaction between the factors. A significant and generalized coherence reduction was observed during the task execution time. Coherence was also diminished at the left frontal cortex and contralateral hemisphere of the utilizing limb (comparing to the right frontal cortex). In conclusion, these findings suggest a certain specialization of the neural circuit, also according to previous investigations. The inter-coherence reduction suggests a spatial inter-electrode dependence during the task, rather than a neuronal specialization. PMID- 17420830 TI - Traumatic subdural hygroma: five cases with changed density and spontaneous resolution. AB - Thirty-four consecutive adult patients with subdural traumatic hygroma were analysed for clinical evolution, serial computed tomography scan (CT), and magnetic resonance imaging (MRI) over a period of several months. Five of the patients presented CT scan and MRI evolution data showing increasing density over a period of 11 days to 6 months post trauma. In these five patients, final clinical and CT scan data were benign, with complete spontaneous resolution. Descriptions in literature of evolving traumatic subdural hygroma have presented CT scan density modifications changing into chronic subdural hematoma. Our patients show another possibility, density transformation, which sometimes show as subdural hematoma in CT scan and MRI, but with final evolution where clinical condition and CT scan return to normal. PMID- 17420831 TI - Duchenne and Becker muscular dystrophy: a molecular and immunohistochemical approach. AB - Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused by mutations in the dystrophin gene. We studied 106 patients with a diagnosis of probable DMD/BMD by analyzing 20 exons of the dystrophin gene in their blood and, in some of the cases, by immunohistochemical assays for dystrophin in muscle biopsies. In 71.7% of the patients, deletions were found in at least one of the exons; 68% of these deletions were in the hot-spot 3' region. Deletions were found in 81.5% of the DMD cases and in all the BMD cases. The cases without deletions, which included the only woman in the study with DMD, had dystrophin deficiency. The symptomatic female carriers had no deletions but had abnormal dystrophin distribution in the sarcolemma (discontinuous immunostains). The following diagnoses were made for the remaining cases without deletions with the aid of a muscle biopsy: spinal muscular atrophy, congenital myopathy; sarcoglycan deficiency and unclassified limb-girdle muscular dystrophy. Dystrophin analysis by immunohistochemistry continues to be the most specific method for diagnosis of DMD/BMD and should be used when no exon deletions are found in the dystrophin gene in the blood. PMID- 17420832 TI - Influence of bromazepam on cortical interhemispheric coherence. AB - Benzodiazepines are among the most commonly prescribed medications due to their therapeutic efficacy in reducing anxiety and inducing sleep. Consequently, they have been widely employed in the pharmacological treatment of several disorders. Nevertheless, few studies have analyzed the effects of bromazepam in electroencephalographic activity (EEG). The present study aimed at investigating the modulatory effects of this drug on brain dynamics. Specifically, the effects of bromazepam (3mg) on EEG coherence were tested in a double-blind experiment. The sample, consisting of 10 healthy subjects (5 male and 5 female), was submitted to ten minutes of EEG recording. The electrophysiological measure (coherence) was analyzed across three experimental conditions: bromazepam, placebo 1, and placebo 2. Results indicate that bromazepam significantly increases cortical interhemispheric coherence. PMID- 17420833 TI - Toxic myopathies: muscle biopsy features. AB - Several drugs and toxic substances can cause muscular abnormalities and are frequent causes of acquired myopathies. We present a series of 32 patients, predominance of young adult patients, diagnosed with toxic myopathy. The most common substances inducing myopathy were corticosteroids (56.2%) followed by the propoxyphene, neuroleptics, zidovudine and drug-induced hypokalemia. The investigation showed normal serum creatine kinase levels in 65.4%, myopathic pattern of the needle electromyography in 40% and the more frequent histological diagnosis of the muscle biopsy was type 2 fiber atrophy (59.3%). Clinical features, etiology, course of the disease, serum levels of muscular enzymes, electromyographic features and, especially, muscle biopsy features are discussed. PMID- 17420834 TI - Anatomopathological aspects of neurocysticercosis in autopsied patients. AB - The aim of this paper was to describe the occurrence and morphology of neurocysticercosis (NCC) in autopsies. We revised 2218 autopsies performed at the School Hospital from Federal University of Triangulo Mineiro, 1970-2003. Data referring to age, gender and color of patients were reported and NCC was microscopically and macroscopically analyzed. We found 53 (2.4%) NCC cases. The mean age was 50 years old, 34 (64.1%) individuals were male and 36 (67.9%) white. Macroscopically, 17 cysticerci were analyzed. The most frequent location was meningocortical in 12 (70.6%) cases. Microscopically, the cysticerci presented an ovoid shape, containing the larvae preserved in 4 (23.5%) cases or in destruction degrees in 13 (76.5%) cases. Therefore, in NCC was found several general pathologic processes (necrosis, interstitial deposits, fibrosis, gliosis, inflammation) amongst which are highlighted beta-fibrillose in 13 (76.5%) cases associated to inflammatory process in 16 (94.1%) cases caused by the parasite, not yet related to NCC, and calcification present in viable and destruction parasites. PMID- 17420835 TI - [The insula and the central core concept]. AB - The characterization of well defined and circumscribed brain regions is particularly useful for the neurosurgical practice once it enhances the tridimensional understanding of its structures and related lesions, and because it induces the development and the utilization of more standard microneurosurgical approaches. In this direction, it is noteworthy that each cerebral hemisphere harbors an evident central core constituted externally by the insula, internally by the basal ganglia and the thalamus, and with the internal capsule within. With a biconvex configuration when seen from above, and located between the sylvian cistern and the supratentorial ventricular cavities, morphologically this central core resembles a head of each brainstem half top, covered by the neocortical mantle of its hemisphere. The central core is attached to the rest of the cerebral hemisphere by isthmi constituted by the different internal capsule fibers. Anteriorly and under the anterior limiting sulcus of the insula there are fibers of the internal capsule anterior limb, superiorly and under the superior limiting sulcus there are the rest of the anterior limb fibers, and the knee and posterior limb fibers that harbors the corticonuclear and the corticospinal tracts, and inferiorly and under the insular inferior limiting sulcus there are the sub- and the retrolentiform internal capsule fibers that enclose the auditory and the optic radiations. Laterally the central core is composed by the insular surface that resembles a shield of the main cerebral subcortical structures. The options of microneurosurgical approaches to the central core related lesions should consider particularly their relationships with the thalamus and with the internal capsule fibers. PMID- 17420836 TI - [The pterional craniotomy: step by step]. AB - This article intends to describe in a didactical and practical manner the frontotemporosphenoidal craniotomy, that is usually known as pterional craniotomy and that constitute the cranial approach mostly utilized in the modern neurosurgery. This is then basically a descriptive text, divided according to the main stages involved in this procedure, and that describes with details how the authors currently perform this craniotomy. PMID- 17420837 TI - [Decompressive craniectomy for treatment of intracranial hypertension secondary to large ischemic cerebral infarction: analysis of 34 cases]. AB - The large ischemic cerebral infarction (LICI) is a blood supply loss of a large area in the brain, mainly on the middle cerebral artery. Is possible that evolutes a major edema, intracranial hypertension and death in about 80% of the cases. OBJECTIVE: To evaluate the results of a decompressive craniectomy on the treatment of the secondary intracranial hypertension to LICI, comparing to other results of medical literature already published. METHOD: Were analysed 34 patients diagnosed with LICI clinically treated unsuccessfully that needed further on the decompressive craniectomy treatment, for the control of intracranial hypertension. RESULTS: 8 patients (23.52%) died, 26 (76.47%) survived, and 2 (7.70%) developed a vegetative state condition. CONCLUSION: The factors age over 50 years and male gender were associated with a high death risk. The level of consciousness at admission and bone flap area were nearer the values of statistic significance. PMID- 17420838 TI - [Treatment of West syndrome with vigabatrin: clinical and electroencephalographic evaluation of 13 patients]. AB - We evaluated the efficacy of vigabatrin (VGB) as a first drug to be used as monotherapy for West syndrome (WS), its side effects and correlations with the electroencephalogram (EEG). The sample consisted of 13 infants examined between October 2001 and September 2002 at IMIP ambulatory patients office or private clinic. Administration of vigabatrin was around 118 mg/kg/day. Suppression of spasms was obtained in 4 children (31%), partial control in 3 (23%), 5 of them did not present therapeutic response (38%) and just one (8%) got worse. On the two patients with tuberous sclerosis, one was seizure-free and in another there was partial control. Side effects happened in 8 children (62%) and consisted of irritability, insomnia, somnolence and agitation, and all of them have been well tolerated. The second EEG showed disappearance of hipsarrhythmia in 6 patients (46%). Four of these were seizure-free. We conclude that VGB is effective and well tolerated as initial monotherapy for WS. PMID- 17420839 TI - [Psychotic syndrome developing into dementia as a clinical manifestation of mitochondrial DNA deletion]. AB - The manifestations of mitochondrial disease are variable, affecting more frequently the organs with high aerobic metabolism in which they are more abundant, for example the nervous system. The beginning of symptoms in general is observed at childhood, but some patients presented on adult age. We present an atypical case associated with mitochondrial DNA deletion. A 39-years-old man with psychiatric symptoms that configured initial clinical picture and only after 12 years of the beginning of symptoms neurological alterations became noticeable. The diagnosis of mitochondrial illness was confirmed by muscle biopsy being documented mitochondrial DNA deletion. PMID- 17420840 TI - [Ventriculoperitoneal shunt infections in children and adolescents with hydrocephalus]. AB - OBJECTIVE: To determine frequency, etiology, site and clinical and laboratory findings of ventriculoperitoneal shunt (VPS) infections in children and adolescents with hydrocephalus managed in Hospital Governador Joao Alves Filho, Aracaju SE, Brazil. METHOD: A non-controlled prospective observational study comprising 50 patients that underwent VPS (58 procedures) from January/2003 to October/2004. RESULTS: Infection rate per procedure was 27.6%; surgical risk index (NNISS-CDC) 0 and 1-2 were 25.7% and 30.4% respectively; surgical site infection was the main complication with 50% of the cases. CONCLUSION: Infection rates per procedure, per patient, and per surgical risk index were high. No statistical differences were found related to the following: age, etiology of hydrocephalus, type of procedure, pre-operative length of stay, duration of procedure, antibiotic prophylaxis, previous central nervous system catheter, and surgical risk index. PMID- 17420841 TI - [Epidemiological and clinical profile of neurocysticercosis patients assisted by the Hospital Universitario Regional de Maringa, Parana, Brazil]. AB - The aim of this study was to drawn an epidemiological and clinical profile of the neurocysticercosis (NCC) patients assisted by Hospital Universitario Regional de Maringa. A transversal, retrospective and descriptive study based on 6,100 records of the Neurology, Neuropediatry, Neurosurgery and Psychiatry clinics was done from January/1998 to December/2004. Of these, 48 were selected with diagnosis of NCC. The prevalence was greater in female (p<0.001), 31-60 years (43.7%), and urban zone (93.8%). The number of NCC cases overcame those notified to the Epidemic Surveillance in 1998, 1999, 2001, 2002 and 2004. The epilepsy due to NCC (CI=1.77;16.07; p=0.0024) was generalized and of late onset in 91.7% patients, being associated to headache in 37.5% ones. There was need of hospitalization in 22.9% of the patients (medium time of 3.91+/-3.35 days). It was concluded that the epidemiological and clinical profile observed is peculiar, displaying some common points with other Brazilian studies. PMID- 17420842 TI - [Electromyografic patterns predictive of motor evolution in neuroschistosomiasis]. AB - Neuroschistosomiasis in myeloradicular pattern is frequently observed in patients from Northeast of Brazil. Despite of this, the evolution of neurologic and electromyographic patterns is not well studied in this group of patients. The aims of this study were to describe and compare the clinic and electromyographic abnormalities of patients with neuroschistosomiasis and radicular involvement. We analyzed 21 electromyographic exams of the lower limbs carried out in the initial presentation of the disease. Electromyographic pattern of 95.2% of patients was compatible to axonal lumbosacral multirradiculopathy, with variable denervation extension, but predominantly from levels L2 to S2. There was variable degree of deficit in the lower limbs, and the follow-up of motor disturbances was better more frequently when the roots were involved in smaller number. Electromyography should permit to know the motor prognosis of patients with myeloradiculitis due schistosomiasis. PMID- 17420843 TI - [ADHD prevalence in four Brazilian public schools]. AB - OBJECTIVE: To define the prevalence of attention-deficit/hyperactivity disorder (ADHD) in children from four Brazilian public elementary schools. METHOD: Study population consisted of all students from the first through fourth grades, age range 6-12 years, who attended four public elementary schools (CIEPs). This prevalence study comprised two steps. During the first step, school teachers screened their own pupils for ADHD using diagnostic criteria from the Diagnostic and Statistic Manual of Mental Disorders - Fourth Edition (DSM-IV). Screening resulted in two groups of children: suspects and non-suspects. In the second step, parents of suspect children were invited for an interview with the researchers, during which they filled ADHD symptoms questionnaire, and in addition a complete history, pediatric physical exam, and neurological exam were performed. At the end of this step, students were classified as "cases" or "undetermined", i.e., those who partially met ADHD diagnostic criteria. RESULTS: From a population of 602 students, 461 were recruited. Considering all four elementary schools, ADHD prevalence was 13%. Male to female ratio was 2:1. The most frequent ADHD subtype was the combined one, accounting for 61.7% of all cases. CONCLUSION: ADHD prevalence in a sample of school-aged children (13%) was higher than the rate that traditionally has been mentioned (3-5%). Boys were more frequently affected than girls and the most prevalent ADHD subtype was the combined subtype, and the latter two findings are concordant with previous studies. PMID- 17420844 TI - [Effects of methylphenidate in auditory processing evaluation of children and adolescents with attention deficit hyperactivity disorder]. AB - PURPOSE: To compare the performance of a group of children and adolescents diagnosed with attention deficit hyperactivity disorder (ADHD) pre and post methylphenidate use in a behavioral auditory processing test battery (AP). METHOD: Twenty-nine subjects, male and female, ranging from 7 to 15 years old have undergone different behavioral auditory processing tests. A control group composed of 29 subjects with and without learning disabilities was also evaluated. RESULTS: The group with ADHD had a performance similar to the control group without learning disabilities which improved after medication. The group with learning disabilities and without ADHD had the worst performance in tests while the group without learning disabilities and without ADHD exhibited the best ones. CONCLUSION: The AP battery was unable to distinguish ADHD patients from paired controls; the use of methylphenidate improved the performance on AP tests of ADHD group in the post-medication evaluation. PMID- 17420845 TI - Kleine-Levin syndrome: interface between neurology and psychiatry. AB - We report the first episode of Kleine-Levin (KLS) syndrome in a 17-year-old male. The illness onset, clinical features, neuropsychological evaluation and polysomnographic recording are described. Typical symptoms hypersomnia, hyperphagia and sexual disinhibition were observed besides behavioral disturbances, polysomnographic and neuropsychological alterations. Behavioral disturbances similar to a manic episode including psychotic symptoms were relevant. The pharmacologic treatment included lithium, methylphenidate and risperidone. The introduction of risperidone aimed the control of psychotic symptoms and the persistent manifestations of hypersexuality after sleepiness control and to the best of our knowledge there are no other reports regarding risperidone use for KLS in the literature. PMID- 17420846 TI - A case of pervasive developmental disorder with chromosomal translocation 1-4. AB - We describe a 5-year-old girl with pervasive developmental disorder associated to chromosome 1-4 translocation, an association that has not been described in specialized literature until this moment. PMID- 17420847 TI - Menkes' disease: case report. AB - Menkes disease is a rare neurodegenerative disorder due to an intracellular defect of a copper transport protein. We describe a 7 months male patient who presented with seizures, hypoactivity and absence of visual contact. The investigation disclosed pilli torti and thrycorrexis nodosa in the hair, low serum levels of both copper and ceruloplasmin, brain magnetic resonance study showed atrophy and white matter hypointensities on T1-weighted images, electroencephalogram reveals moderate background activity disorganization and epileptiform activity, and muscle biopsy with type 2 fiber atrophy. The clinical, laboratorial, genetic, muscle biopsy and neurophysiological findings in Menkes disease are discussed. PMID- 17420848 TI - Hypomyelination with atrophy of the basal ganglia and cerebellum: case report. AB - Hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC) is a rare disease that has been recently described. It must be remembered as a possible etiology of leukoencephalopathies in children. We describe a typical case of H-ABC in a 11-month-old boy. He presents with global development delay, oral dyskinesia, and global dystonia and spasticity. Magnetic resonance imaging disclosed typical features of H-ABC and clinical laboratory tests were all negative. A slow neurological deterioration has been detected with worsening of involuntary movements. PMID- 17420849 TI - Probable causal link between epilepsy and sleep apnea: case report. AB - Patients with epilepsy were reported to have concomitant sleep apnea, but it has been rarely linked to the epilepsy itself. We present a case of a 28-year-old, obese man with secondary medically resistant partial complex epilepsy due to a brain trauma, with progressive snoring, and sleep agitation, apneas, and important daytime somnolence. It was noticed in the polysomnographic study that he had several sleep respiratory events, probably due both to the epileptic seizures and the sleep apnea syndrome as a co-morbidity. Apnea and epilepsy will be discussed. A careful video-EEG-polysomnography study is important in evaluating refractory epileptic patients and/or epileptic patients with snoring. PMID- 17420850 TI - Merosin-positive congenital muscular dystrophy: neuroimaging findings. AB - Congenital muscle dystrophy (CMD) is a heterogeneous group of autosomal recessive myopathies. It is known that CMD may affect the central nervous system (CNS). Some authors have shown that merosin-negative CMD patients may have encephalic metabolic disturbances. In order to study metabolic changes within the brain, the authors performed a magnetic resonance spectroscopy (MRS) study in a 1-year-old girl with merosin-positive CMD (MP-CMD). MRS of brain demonstrated that NAA/Cr ratio was decreased (1.52), while Cho/Cr ratio was increased (1.78). These findings suggest that metabolic changes in CNS can also be found in patients with MP-CMD. PMID- 17420851 TI - Dermoid cyst of the anterior fontanelle in adults: case report. AB - Head and neck dermoid cysts are relatively rare, and usually occur during childhood as solitary lesions. They are often identified and surgically removed at birth, being uncommon in adults. A 23-year-old male presented with a congenital tumor of the anterior fontanelle, which histopathological examination revealed a dermoid cyst. Surgical intervention is the treatment of choice to remove this lesion. The objective of this study is to report the case, once this type of lesion is rare in adults. PMID- 17420852 TI - Dandy-Walker syndrome in adult mimicking myasthenia gravis. AB - The Dandy-Walker syndrome (DWS) is a rare posterior fossa malformation. The DWS can occur associated with other brain or systemic malformations, but ocular abnormalities in this disease are rare and clinical findings mimicking myasthenia gravis have not been described to date. We report a 23-year-old woman who presented mild limitation of the ocular movements with progressive palpebral ptosis, which changed in intensity during the day. The investigation showed negative anti-acetylcholine receptor antibody, repetitive nerve stimulation and "Tensilon test", but the brain magnetic resonance image reveals DWS with hydrocephalus associated with callosal dysgenesis. The characteristic of disease, clinical manifestations and pathologic features, specially the clinical evaluation of ocular abnormalities in suspicion of DWS, including the MG in differential diagnosis are discussed. PMID- 17420853 TI - Isolated Pathetick nerve paresis by compression from a dolichoectatic basilar artery: case report. AB - The article describes a woman with 53 year-old that has presented diplopia when looking down and adopted a tilted head position in order to read for the last six months. The neuro-ophthalmic examination shows isolated right superior oblique muscle paresis. The magnetic resonance and the angioresonance show the dolicoectatic basilar artery compressing the right ventral lateral brainstem. PMID- 17420854 TI - Comparison study of executive functions in Parkinson's disease and degenerative cerebellar disease's patients. PMID- 17420855 TI - Theory of mind and the frontal lobes. PMID- 17420856 TI - Biological aspects of Autoplusia egena (Guenee) (Lepidoptera: Noctuidae, Plusiinae). AB - Autoplusia egena (Guenee) is a moth distributed throughout the Western hemisphere. The larvae have already been found feeding on several different plant families, including important crops such soybeans and beans. To contribute to the knowledge of its biology in laboratory conditions, and considering the duration and the morphometry of each development stage, a laboratorial rearing was accomplished at 25 +/- 1 degrees C, 70 +/- 10% RH and 14h photophase. The larvae were fed with black bean leaves and the adults were fed with a honey solution at 10%. The results, expressed by the mean (+/- SE), for the periods of each stage were: egg 3.0 +/- 0.00 d; larva 15.7 +/- 1.25 d; pre-pupa 1.9 +/- 0.05 d; pupa 8.8 +/- 0.09 d and adult: longevity 12.1 +/- 0.95 d, pre-oviposition 5.4 +/- 0.50 d, oviposition 6.3 +/- 1.10 d and post-oviposition 1.4 +/- 0.87 d. The larvae went through five instars, for which the mean width of the cephalic capsules were 0.302 +/- 0.001 mm; 0.500 +/- 0.003 mm; 0.854 +/- 0.008 mm; 1.424 +/- 0.011 mm and 2.744 +/- 0.053 mm. The average length, width, and weight of the pupae were 16.965 +/- 0.003 mm, 4.674 +/- 0.040 mm and 0.217 +/- 0.003 g, respectively. PMID- 17420857 TI - [Fauna of Scarabaeidae-Laparosticti and Trogidae (Coleoptera: Scarabaeoidea) associated with Cloud Forest, shaded coffee plantations and transformed communities in Central Veracruz, Mexico]. AB - Beetles were collected from April 2002 to July 2003 from the following sites in the centre of the state of Veracruz located every 200 m along an altitudinal gradient (1000 - 1400 m asl): three fragments of cloud forest, three shaded coffee plantations, an open canopy coffee plantation, secondary forest, and a pasture. A total of 9,982 specimens were captured, belonging to the families Scarabaeidae and Trogidae, and representing 21 genera and 50 species. The genera Ataenius Harold, Onthophagus Latreille and Aphodius Illiger represented 48% of the species of Scarabaeidae. Species richness was found to decrease with increasing altitude; there were 36 species at 1000 m asl, 27 species between 1200 and 1300 m asl, and 26 species at 1400 m asl. Abundance along the altitudinal gradient follows a pattern of few abundant species and many species with few specimens. Beetle activity is related to precipitation. In the nine communities studied, species richness was observed to increase when precipitation was greater than 100 mm at the beginning of the rainy season. On the landscape scale, there were 44 species during the rainy season, 22 during the windy nortes season, and 24 during the dry season. In terms of trophic guild, 40% of the beetles captured were saprophagous that feed on decomposing organic material from plants and 60% were saprophagous that feed on decomposing organic material from animals (30% necrophagous, 26% coprophagous, 4% telio-necrophagous). PMID- 17420858 TI - Life history of Aneugmenus merida Smith (Hymenoptera: Tenthredinidae) in the Venezuelan Andes. AB - Details of the life cycle of Aneugmenus merida Smith (Hymenoptera: Tenthredinidae) reared under laboratory conditions are reported. Females may produce parthenogenetic or non-parthenogenetic eggs. After eclosion, males of A. merida go through five larval instars, pre-pupa, pupa and imago. The complete life cycle lasts approximately 44 days. Larvae feed on bracken [Pteridium caudatum (L.) and Pteridium arachnoideum (Kaulf)] (Dennstaedtiaceae). We also report on morphological observations of the sinus sexualis in males by scanning electron micrographs and give details on mating behavior. PMID- 17420859 TI - [Effects of environmental factors on the ant fauna of restinga community in Rio de Janeiro, Brazil]. AB - The effects of environmental factors on the richness, diversity and abundance of ants were studied in the Restinga da Marambaia, south coast of Rio de Janeiro State, Brazil. The samples were taken using pitfall traps in August/2004 (winter) and March/2005 (summer) in three different vegetation types: (1) herbaceous ridge palmoid (homogeneous habitat); (2) shrub dune thicket and (3) ridge forest (heterogeneous habitats). At each habitat a range of environmental attributes was recorded: soil temperature and humidity, percentage of soil covering by litter and litter depth. Ninety-two ant species belonging to 36 genera and eight subfamilies were recorded. Density of ant species and abundance varied significantly between habitats and seasons; ant diversity varied only between habitats. Homogeneous habitat had lower ant species density, abundance and diversity than heterogeneous habitats. The two first variables were positively correlated with litter depth and both were higher in summer than in winter samples. There were more species of Ponerinae and Ectatomminae in heterogeneous than in the homogeneous habitat, whereas the Formicinae species were more abundant in the later. PMID- 17420860 TI - [Harvesting dynamics of pollen sources by Melipona scutellaris Latreille (Hymenoptera: Apidae): a comparative analysis with Apis mellifera L. (Hymenoptera: Apidae) in the Atlantic Forest Domain]. AB - Generalist foraging behavior among stingless bees is accepted but untested, as well as the subsidiary hypothesis of floral preferences in the genus Melipona. Here we analyzed those hypotheses comparing the use of floral sources of pollen, through paired analyses of pollen samples from different colonies of Melipona scutellaris Latreille, in three areas of the Atlantic Forest Domain, in Northern Brazil. From August, 2004 to January, 2005, monthly samples of pollen were collected at the entrance of twelve colonies of M. scutellaris. In two places, four colonies of M. scutellaris were compared with four colonies of africanized Apis mellifera L. The main pollen sources chosen by the colonies of M. scutellaris were flowers of the following plant families, in decreasing order of importance: Myrtaceae, Mimosaceae, Anacardiaceae, Sapindaceae, Caesalpiniaceae and Fabaceae. Productive pollen sources of Asteraceae, Arecaceae e Rubiaceae were heavily exploited by the colonies of A. mellifera and discharged by the colonies of M.scutellaris. Often, both species shared the main productive pollen sources, as the flowers of Myrtaceae and Mimosaceae. On the other hand, no pollen sources were heavily exploited altogether by both of them, as a rule. In different places and periods, the colonies of M. scutellaris presented high intra-specific similarity and they formed distinct clusters apart from A. mellifera. Therefore, the selection of pollen sources by colonies was species dependent. The paired comparisons refute the hypothesis of random flower exploitation by colonies and give support to the subsidiary hypothesis of selectivity or floral preferences by M.scutellaris. PMID- 17420861 TI - Three new species of Peristicta Hagen in Selys (Odonata: Zygoptera: Protoneuridae) from Brazil. AB - Three new species of Peristicta Hagen in Selys from Brazil are described and illustrated: P. janiceae from Minas Gerais State (Diamantina, Gouvea, Lagoa Santa, Serra do Caraca, Serra do Cipo, Urobotanga), P. jalmosi from Goias State (Chapada dos Veadeiros, Reserva da Universidade de Brasilia) and Minas Gerais State (Urobotanga, Lagoa Santa, Ponte Nova, Sao Joao del Rey,) and P. muzoni from Mato Grosso State (Serra da Bodoquena). An identification key for males of Peristicta is presented. PMID- 17420862 TI - Taxonomic survey of Drosophilidae (Diptera) from mangrove forests of Santa Catarina Island, Southern Brazil. AB - Assemblages of drosophilids have been characterised in several environments of the Brazilian territory, like the Atlantic Rain Forest, urban areas, cerrados, the Amazon Forest, and others. The present survey is the first attempt to characterise the fauna of Drosophilidae in mangrove forests, an environment typical of tropical coasts worldwide. Twenty-eight samples were collected from the three main mangrove forests of Santa Catarina Island, southern Brazil, using banana-baited traps hung in trees. Samples were taken in January (summer), April (autumn), July (winter) and October (spring) between July 2002 and July 2005. In total, 82,942 specimens of drosophilids were caught, belonging to 69 species of six genera - Amiota Loew, Drosophila Fallen, Leucophenga Mik, Scaptodrosophila Duda, Zaprionus Coquillett and Zygothrica Wiedemann. The high abundance of D. simulans Sturtevant was remarkable, with some notable peaks of D. malerkotliana Parshad & Paika in autumn samples. Other common species were Zaprionus indianus Gupta, D. mediostriata Duda and D. willistoni Sturtevant. We also collected 45,826 flies of family Curtonotidae, the sister-group of Drosophilidae virtually absent in other environments. The assemblages of drosophilids were very similar in the three mangrove forests surveyed, despite the different surrounding environments. In general, the species sampled in the mangroves were the same as those observed in the surrounding environments, but in varying abundances. This suggests that drosophilids are differently affected by environmental pressures operating in mangroves. PMID- 17420863 TI - [Effect of Cu2+ and Zn2+ ions in Ca-ATPase activity isolated from Pachymerus nucleorum (Fabricius) (Coleoptera: Chrysomelidae, Bruchinae) larvae]. AB - ATPases, an important target of insecticides, are enzymes that hydrolyze ATP and use the energy released in that process to accomplish some type of cellular work. Pachymerus nucleorum (Fabricius) larvae possess an ATPase, that presents high Ca ATPase activity, but no Mg-ATPase activity. In the present study, the effect of zinc and copper ions in the activity Ca-ATPase of that enzyme was tested. More than 90% of the Ca-ATPase activity was inhibited in 0.5 mM of copper ions or 0.25 mM of zinc ions. In the presence of EDTA, but not in the absence, the inhibition by zinc was reverted with the increase of calcium concentration. The inhibition by copper ions was not reverted in the presence or absence of EDTA. The Ca-ATPase was not inhibited by treatment of the ATPase fraction with copper, suggesting that the copper ion does not bind directly to the enzyme. The results suggest that zinc and copper ions form a complex with ATP and bind to the enzyme inhibiting its Ca-ATPase activity. PMID- 17420864 TI - A regional analysis of weather mediated competition between a parasitoid and a coccinellid predator of oleander scale. AB - The regulation of an asexual population of the oleander scale [Aspidiotus nerii Bouche (Hemiptera: Diaspididae)] on California bay tree [Umbellularia californica (Hopk. & Arn.) Nut.] by two natural enemies; an idiobiont, ectoparasitoid Aphytis chilensis Howard (Hymenoptera: Aphelinidae) and a coccinellid predator (Rhysobius lophanthae (Blaisd.) (Coleoptera: Coccinellidae), was examined using a general weather-driven, tri-trophic, physiologically based age-mass structured demographic model. The model is of intermediate complexity and was parameterized using extensive laboratory data and field observations from Albany, CA. Temperature-dependent physiological indices were estimated from the laboratory data and used to scale per capita growth, fecundity and survivorship rates from maximal values in a time varying environment. The tri-trophic model was integrated in a GIS (geographic information system) and the species dynamics examined across years and across the ecological zones of California. Field data and simulation results suggested the coccinellid predator was the most important regulating agent of oleander scale in the mild climate of Albany. However, multiple linear regression analysis of simulation data across all ecological zones of California shows that the parasitoid A. chilensis is the most important agent in suppressing oleander scale densities in warmer climates, while the predator R. lophanthae increases scale density an average of 9.7% across all regions. PMID- 17420865 TI - [Parasitism capacity of Trichogramma pratissolii Querino & Zucchi (Hymenoptera: Trichogrammatidae) on alternative hosts, under different temperatures]. AB - The successful use of Trichogramma as biocontrol agent depends on its mass production in laboratory, a fundamental step for any biological control program among other factors. This work investigated the parasitism capacity of Trichogramma pratissolii Querino & Zuchi (Hymenoptera: Trichogrammatidae), a new recorded Trichogramma species, parasitizing eggs of Anagasta kuehniella (Zeller) and Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) under the temperatures of 15, 18, 21, 24, 27, 30 and 33 degree Celsius. Eggs of these hosts were offered to newly emerged females during 24h. This procedure was repeated daily for each female and each temperature up to female death, in order to estimate daily and accumulated parasitism, and female longevity. On both hosts, the daily parasitism decreased as function of the female age. Under all temperatures studied and both hosts the highest rate of parasitism was observed during the first 24h of host exposure, and reached 80% of total parasitism in the 4th and 3rd days when parasitizing A. kuehniella and C. cephalonica, respectively. On both hosts, the highest parasitism rate was observed under temperatures from 21 degree Celsius to 27 degree Celsius. Average longevities of T. pratissolii females deprived of food emerging from A. kuehniella and C. cephalonica lived for 1.0 and 8.9 days when reared at 15 degree Celsius e 33 degree Celsius, respectively. The results indicate that eggs of A. kuehniella and C. cephalonica and temperatures from 21 degree Celsius to 27 degree Celsius were appropriate to rear T. pratissolii. PMID- 17420866 TI - [Diversity of insects captured by weaver spiders (Arachnida: Araneae) in the cocoa agroecosystem in Tabasco, Mexico]. AB - The aim of this work was to know the diversity of insects captured by weaver spiders in a plantation of cocoa (Theobroma cacao L.) of 6 ha in the State of Tabasco, Mexico. The study was carried out from July 2004 to June 2005 by means biweekly samples of the insects captured on the spiders webs. The total of 3,041 webs of 54 species of spiders belonging to seven families (Araneidae, Theridiidae, Tetragnathidae, Uloboridae, Pholcidae, Dyctinidae and Linyphiidae) were revised. We found 1,749 specimens belonging to 10 orders of insects, represented by 93 families, the majority of Coleoptera, Diptera and Hemiptera that constituted 74% of the identified families. The biggest number of specimens of all orders was captured by Araneidae, except of Isoptera, whose specimens were captured mainly by the family Theridiidae. The index of diversity (H'), evenness (J') and similarity (Is), applied to know the diversity of families of insects captured among families of spiders, varied from 0.00 to 3.24, 0.00 to 0.81, and 0.04 to 0.522, respectively. We conclude that there is a wide diversity of insects predated by the weaver spiders in the cocoa agroecosystem, and that there are species that can be promising for the biological control of pests. PMID- 17420867 TI - [Biological parameters and damage by Macrosoma tipulata Hubner (Lepidoptera: Hedylidae), in cupuacu tree [Theobroma grandiflorum (Wild ex Spreng Schum)] in Amazonas, Brazil]. AB - Biological aspects of the caterpillar Macrosoma tipulata Hubner and its damage to young leaves of Theobroma grandiflorum (Wild ex Spreng Schum) were studied under laboratory conditions of 24.5 +/- 0.7 masculineC, UR 61.6 +/- 9.1% and 12h photophase. The larval stage presented five instars with a total duration of 15.9 +/- 4.31 days, and survival of 98%. The pupal period stage was 7.0 +/- 0.56 days for males and 6.9 +/- 0.56 days for females, with 99% survival for both sex. The pupal mean length was 18.9 +/- 0.89 mm for males and 19.1 +/- 1.13 mm for females. The mean longevity of the adult was 10.3 +/- 2.25 days. The vertical distribution of damage was relatively uniform, with a tendency of greater defoliation in the middle of the treetop, but without significant difference among the three positions. PMID- 17420868 TI - [Population dynamics and vertical distribution of Scaptocoris carvalhoi Becker (Hemiptera: Cydnidae) in pastures]. AB - Studies on population dynamics and vertical distribution in the soil of Scaptocoris carvalhoi Becker were carried out in a pasture area in Parauna, State of Goias, Brazil, from November 2004 to October 2005. One meter-deep holes were excavated in the soil, except on the season of lower precipitation, when 2-meter holes were excavated (June, August and September 2005). On each evaluation, the specimens were collected and grouped according to the depth they were found in the soil profile (0-20, 21-40, 41-60, 61-80 and 81-100 cm). Nymphs occurred during the entire length of the study and were more abundant than adults. Adult population augmented with the increase of rainfall. Nymphs and adults were located in the top layer of the soil during the rainy season and were found deeper in the soil during droughts. However, adults were less tolerant to hydric stress and burrowed deeper into the soil at the beginning of the dry season, while nymphs burrowed deeper only at the end of the season. Results of this study are in accordance to other studies on various species of Scaptocoris, in other regions of Brazil. PMID- 17420869 TI - [Susceptibility of Acarophenax lacunatus (Cross & Krantz) (Prostigmata: Acarophenacidae) to Sulfur]. AB - The parasitic mite Acarophenax lacunatus (Cross & Krantz) is a potentially important biocontrol agent of stored grain insect pests, but its presence in insect mass rearing is undesirable. This study was carried out to evaluate the susceptibility of A. lacunatus to sulfur. The experimental units were petri dishes containing 30 g of whole wheat grains infested with 30 adults of Rhyzopertha dominica (Fabricius) (Coleoptera: Bostrichidae). The treatments consisted of pulverizing the wheat grains with sulfur, at 0, 0.15, 0.3, 0.6, 0.9 and 1.2 mg a.i./g, in five replicates. Three inoculations of A. lacunatus were carried out in each petri dish at five, ten and fifteen days after infestation with R. dominica. The petri dishes were stored for 60 days at 30 +/- 1 degree Celsius, 60 +/- 5% R.H. and 24h scotophase. The parasitic mite was susceptible to all sulfur doses used in the test. The number of eggs of R. dominica parasitized by A. lacunatus showed a decreasing trend with the increase in doses of sulfur, as did the number of physogastric females of the mite. The mite showed a negative correlation of their instantaneous rate of increase with sulfur dose. No mites developed at sulfur doses higher than 0.3 mg a.i./g. The susceptibility of A. lacunatus to sulfur is an important aspect to consider in the control of this parasitic mite in colonies of R. dominica. In addition, this control method has low cost and is easy to use. PMID- 17420870 TI - [Occurrence and fluctuation of mite populations associated with rubber tree crop neighbor of fragments of cerrado]. AB - The basic knowledge of the seasonal occurrence of mites can supply data for elaboration of programs of ecological management to be implanted with success in the future. The objective of this study was to determine the species richness and the seasonality of mites present in two areas of rubber tree crops neighboring to native areas in Itiquira, MT. Along one year, 25 quantitative samplings were accomplished in rubber tree crop neighboring to two fragments of Cerrado (Cerradao and Mata Riparia). There were registered 199,380 mites, of 48 species, belonging to 15 families. Of those, 13 species are phytophagous, 18 predators and 17 mycophagous or of unknown alimentary habit. Three phytophagous species represented more than 97% of the mites collected: Phyllocoptruta seringueirae Feres (80.8%), Tenuipalpus heveae Baker (12.7%) and Calacarus heveae Feres (3.6%). Among the predators, the most abundant species were Agistemus sp., Scirula sp. and Euseius concordis (Chant). Twenty-eight species were common to both crops. The families that had the largest number of species collected in the neighboring area to Cerradao were Tydeidae (7), Tarsonemidae (6), Eriophyidae and Phytoseiidae (4), and in the area close to Mata Riparia, Tydeidae (9) and Phytoseiidae (8). The presence of vegetation near the crop should explain the great number of species of mites classified as accidental found in this study. The largest abundances and species richness occurred in the end of the rainy season and beginning of the dry season. PMID- 17420871 TI - [Phlebotomine (Diptera: Psychodidae) and leishmaniasis in Rio Grande do Norte State Brazil: anthropic environment responses]. AB - Natural environmental changes or those resulting from anthropic factors and their impact on infectious diseases have been evaluated in several studies. The objective of this work was to analyze the correlation between the anthropic environment, phlebotomine and leishmaniases in Rio Grande do Norte State, in Northeast Brazil. Information relative to the distribution of vector species in visceral and tegumentary leishmaniasis areas was associated to the record of cases notified by Public Health organs. The analysis suggests associations between the vector species and distribution of the disease with demographic and physionomic characteristics, disorderly growth in the metropolitan area, living conditions and environmental degradation of the Eastern Littoral, the principal area of notified visceral leishmaniasis cases. PMID- 17420872 TI - [Diversity and abundance of phlebotomine of the genus Lutzomyia (Diptera: Psychodidae) in areas of forest in the northeast of Manacapuru, Amazonas State, Brazil]. AB - The genus Lutzomyia has great importance in the New World, with some species implicated in the transmission of causal agents of leishmaniases, bartonellosis and arboviruses. From April 2003 to June 2004 an investigation was undertaken on the richness and abundance of the sand fly fauna in the northeast area of Manacapuru county, Amazonas State. The captures were carried out, with 16 light traps CDC, in areas of forest known as terra firme along the highway Manuel Urbano. In the period of 13 months we collected a total of 10,446 sandfly specimens, 3,908 males (38%) and 6,465 females (62%), distributed in 43 species belonging to the genus Lutzomyia, 10 subgenera and six species groups. These results evidenced a diversified and abundant sand fly fauna, with some species not yet reported for Manaus county, close to the study area. PMID- 17420873 TI - Do Trichogramma minutum Riley and Trichogramma bennetti Nagaraja & Nagarkatti (Hymenoptera: Trichogrammatidae) occur in Brazil? AB - According to some catalogues, Trichogramma minutum and T. bennetti have been recorded in Brazil. However, the information obtained in studies with Trichogramma in recent years indicate that those species do not occur in Brazil. PMID- 17420874 TI - [Parasitism in Trialeurodes variabilis (Quaintance) (Hemiptera: Aleyrodidae) by Encarsia hispida De Santis (Hymenoptera: Aphelinidae), in papaya, in Brazil]. AB - Infestation of Trialeurodes variabilis (Quaintance) was observed in October 2004, in papaya plants of cultivar Sunrise Solo, under screenhouse conditions, in Cruz das Almas, State of Bahia, Brazil. In infested leaves, around 20% of parasitism on nymphs was verified. Leaves with parasitized nymphs were kept in laboratory until emergence of the parasitoid, identified as Encarsia hispida De Santis. This is the first time that this parasitoid was detected on T. variabilis nymphs in Brazil. PMID- 17420875 TI - [New records of the cryptobiotic ant Creightonidris scambognatha Brown (Hymenoptera: Formicidae) from Brazil]. AB - A queen of Creightonidris scambognatha Brown is recorded for the fist time for the State of Mato Grosso, Pantanal area, and a worker for the State of Bahia, also for the first time for Brazil. Both specimens were obtained with Winkler extractors. PMID- 17420876 TI - [Nesting of Euglossa (Euglossa) melanotricha Moure (Hymenoptera: Apidae) in the ground of cerrado]. AB - Nests of euglossine bees are difficult to found in nature, with descriptions available for only 20% of known species. A nest of Euglossa (Euglossa) melanotricha Moure found inside a cavity in the ground at a Cerrado area in Uberlandia, Minas Gerais state, is described. Comparisons of nesting behavior and social structure with those reported for Euglossa (Euglossa) cordata (L.) and Euglossa (Euglossa) townsendi Cokerell are presented and discussed. PMID- 17420877 TI - The correct identity of a louse sample (Phthiraptera: Menoponidae) from the roadside hawk, Rupornis magnirostris (Gmelin) (Falconiformes: Accipitridae). AB - A report of a louse sample identified as Colpocephalum cholibae Price & Beer by Oliveira et al. (2004), from the roadside hawk [Rupornis magnirostris (Gmelin)] in the state of Sao Paulo, Brazil, is regarded as a misidentification. A correction to the identity of the lice is given as Kurodaia (Kurodaia) fulvofasciata (Piaget). Key morphological differences between the genera Colpocephalum and Kurodaia are discussed, as well as possible reasons for the misidentification. PMID- 17420878 TI - [Tobacco and childhood ]. PMID- 17420879 TI - Does ABPM replace office blood pressure measurement in the clinical evaluation of patients? Point of view--not yet. PMID- 17420880 TI - [ Does the MAPA (Ambulatory Monitoring of Arterial Pressure) modify the diagnosis and treatment of arterial hypertension? Point of view- Yes]. PMID- 17420881 TI - [Burnout syndrome: reality or fiction ?]. PMID- 17420882 TI - [In high-risk patients, submitted to myocardial revascularization, does preoperative chest physiotherapy prevent pulmonary complications ? ]. PMID- 17420884 TI - [Diagnosis of hemorrhoids ]. PMID- 17420883 TI - [Gastric cancer in the elderly: when not to operate?]. PMID- 17420886 TI - [Lung disease associated with acute myeloid leukemia and hyperleukocitosis. ]. PMID- 17420887 TI - [Adolescent females' knowledge about pregnancy prevention methods and sexually transmitted diseases]. AB - OBJECTIVE: To evaluate knowledge about sexuality, contraceptive methods and sexually transmitted diseases (STD) by female adolescents from both rural and urban zone attending public school. METHODS: A cross sectional study was made with 506 teenagers, 10 to 16 years old, attending Dr. Roberto Feijo Public School in Guararema, Sao Paulo. A semi-structured questionnaire with general questions about sexuality, contraceptive methods and STD was administered. The Chi-square test was used to verify the association between variables. RESULTS: Mean age of the girls from the rural zone was 13 years and 11 months and from the urban zone age was 13 years and 7 months, with no statistical difference. Of all the girls, 31% came from the rural and 69% from the urban zone. Adolescents from the rural zone looked for more information about sexuality (81.2%) when compared to those from the urban zone (72.2%) (p<0.0568). Parents were the main source of information for both zones. The condom was the most familiar method in the rural (44%) and the urban (45%) zones (p=0.0022). AIDS was the best known STD by girls from the rural (43%) and urban (39%) zones (p=0.7843). CONCLUSION: Most of the surveyed female adolescents sought information about sexuality, however their knowledge about STD and contraceptive methods was inadequate. PMID- 17420888 TI - [Relation between age at menarche and final height of women in the Pro-Saude Study]. AB - OBJECTIVE: To evaluate the relation between age at menarche and final height. METHODS: Complete data were obtained for 1,940 participants of a cohort study (Pro-Saude Study) of civil servants at a university in Rio de Janeiro. A self administered questionnaire was used to obtain the date of birth, age of the first menstruation and parents' educational level. Height in centimeters was measured twice by trained anthropometrists. Data were analyzed using linear regression models with final height as the dependent variable. RESULTS: After adjusting for parents' educational level, it was observed that for each additional year in the age at menarche there was an increase in the final height of 0.91 cm for women aged 22 to 30 years, 0.44 cm for women aged 31 to 40 years and 0.40 cm for those between 41 and 50 years of age. In contrast, for women of 51 to 60 years of age, each additional year in the age at menarche was associated with a decrease of 0.64 cm in the final height. Focusing only on women between 22 and 50 years of age, no significant interaction was identified between age and age at menarche, however a strong negative confounding effect exerted by participants' age and by their parents' educational level was identified. CONCLUSION: Results of this study suggest that, for younger generations, a later age at menarche is directly associated with final height. Childhood socioeconomic conditions and generation effects can be important confounding factors and should be considered in analyses of this relationship. PMID- 17420889 TI - [Indoor allergen sensitization profile in allergic patients of the allergy clinic in the University Hospital in Uberlandia, Brazil]. AB - OBJECTIVE: To evaluate allergens among patients with allergic respiratory disease attended at the Allergy Clinic of the University Hospital-Federal University of Uberlandia. METHODS: A retrospective study was performed using medical records. Patients were included if their ISAAC questionnaires were correctly filled out and their skin prick tests were positive to at least one of the allergens from Dermatophagoides pteronyssinus (Der p), Dermatophagoides farinae (Der f), Blomia tropicalis (Blo t), Canis familiaris (Can f), Felis domesticus (Fel d), Blattella germanica (Bla g) and Alternaria alternata. RESULTS: Two-hundred and twelve medical records fulfilled the inclusion criteria. Allergic rhinitis was the main clinical diagnosis (32%), followed by concomitant manifestation of asthma and rhinitis (29.7%), and asthma, rhinitis and atopic dermatitis (9.4%). Asthma alone was found only in 1.9% of patients. The total sensitization observed was 73.5%, of which 61.7%, 59.9%, 54.7%, 45.7%, 38.2%, 33.3%, 9.9% were sensitized to Der p, Der f, Blo t, Bla g, Can f, Fel d and Alternaria, respectively. No significant difference was found between allergic disease types and source of allergen sensitization. CONCLUSION: The highest sensitization in allergic patients under study was to dust mites, especially Der p and Der f. It is noteworthy that the number of patients sensitized to cockroach extract was uncommonly high when compared to previous studies. PMID- 17420890 TI - [Effects of streptokinase and hyperbaric oxygen therapy on venous occlusion after limb reimplantation in rats]. AB - OBJECTIVE: The effectiveness of streptokinase and hyperbaric oxygen therapy on venous occlusion after limb reimplantation was tested in rats. METHODS: Amputation with preservation of vessels and nerves of the right hind limb was carried out in 140 rats. Groups MG0, MG1, MG2, MG3 and MG4 were submitted to 0, 1, 2, 3 and 4 hours of venous occlusion. MG3 was elected as control for the experimental groups. Groups EG1 and EG2 were submitted to 3 hours of venous occlusion and were treated with streptokinase and hyperbaric oxygen therapy. Limbs were observed for 7 days and their mortality and survival rates were studied. RESULTS: Trans-operatory mortality rates in groups MG0, MG1, MG2, MG3 and MG4 were 0, 10, 15, 30 and 60% respectively and the postoperative mortality rates were 5; 11.1; 11.7; 14.2 and 100% respectively. The limb survival rates were 100%, 87.5%, 80% and 66.67% respectively and 76.9% and 100% in EG1 and EG2. Model groups were statistically different, except for MG1 and MG2 in trans operatory mortality rates. There were no statistical differences in postoperative mortality rates between model groups except for MG3 and MG4. Model groups were statistically different, with the exception of MG1 and MG2, in limb survival rates. EG1 and MG3 showed no statistical difference in limb survival and EG2 had a better limb survival than MG3. CONCLUSION: Results suggest that the administration of streptokinase does not change effects of venous occlusion and that hyperbaric oxygen therapy may decrease the effects of venous occlusion in limbs. PMID- 17420891 TI - [Bacteremias at a teaching hospital: etiology, antimicrobial susceptibility pattern and risk factors for mortality]. AB - OBJECTIVE: To evaluate the frequency and profile of bacteremia, its antimicrobial susceptibility and to analyze predictors of mortality in bloodstream infections (BSI) at this Teaching Hospital from January 1, 2000 to December 31, 2001. METHODS: DESIGN: retrospective cohort. Patients over one year old with clinically significant episodes of BSI which were microbiologically documented were included in the study. The Cox proportional hazards risk model was applied to identify prognostic factors related to death by bacteremia. RESULTS: A total of 295 episodes of BSI were detected. The most common pathogen was S. aureus: 118 (40.0%), with 55.9% of MRSA. Mortality associated with bacteremia was 34.5%. Independent predictors of mortality were: inadequate initial therapy (HR adjusted 2.05 IC95%: 1.25-3.36) and severity of the clinical manifestations (HR adjusted 5.52 IC95%: 3.15-9.69). CONCLUSION: This study disclosed high mortality rates due to BSI and a high frequency of MRSA. Inadequate initial therapy and severity of clinical manifestations were significantly and independently associated with mortality. PMID- 17420892 TI - [Delay in ophthalmologic examination of diabetic retinopathy patients]. AB - OBJECTIVE: To assess the delay in the ophthalmic examination of patients affected with severe diabetic retinopathy but no visual complaint. METHODS: A transversal analytical study was conducted of 44 visually asymptomatic diabetic patients. All were eligible to retinal panphotocoagulation in at least one eye. Duration and type of the disease as well as time of the first ophthalmic examination were established. RESULTS: Of the 44 patients, 6 (13.7%) had diabetes type 1 and 38 (86.3%) had diabetes type 2. Proliferative diabetic retinopathy was present in 24 (54.5%) and severe or very severe non proliferative retinopathy was present in 20 (45.5%). Best visual acuity ranged from 0.5 to 1.0 (mean 0.83+/-0.17) according to the Snellen chart. The time interval between diagnosis of diabetes and the first ophthalmic examination ranged from 7 to 19.5 years (13.2+/-5.12) for diabetes type 1 and from 3 months to 18 years (5.2+/-4.81) for diabetes type 2. CONCLUSION: Advanced diabetic retinopathy may be detected at different times of the ophthalmic examination in both types of diabetes. Although the mean interval between the diagnosis and the first examination is 13 years for diabetes type 1 and 5 years for diabetes type 2, this severe kind of retinopathy occurred as early as three months after diagnosis of diabetes. This points to the need for an earlier ophthalmic examination in the case of this disease. PMID- 17420893 TI - [Frequencies of blood groups, ABO and Rh D incompatibility in post-delivery women and their liveborn]. AB - OBJECTIVE: This study aimed to assess the frequency of different blood phenotypes and to predict the risk of Rh D alloimmunization and maternal-fetal incompatibility in a Brazilian population living in the West zone of the city of Sao Paulo-Brazil. METHODS: This descriptive study evaluated 2,372 post-delivery women and their liveborn during one year. Blood types were analyzed by means of tube agglutination tests. RESULTS: The blood type frequencies were: 50.67 O, 32.17 A, 13.45 B, 3.75 AB, 90.34 Rh D(+) and 9.66 Rh D(-). ABO maternal-fetal incompatibility was detected in 18.4% and Rh D incompatibility in 7%. CONCLUSION: The fraction of Rh D(-) population at high risk for Rh D alloimmunization was 82%, emphasizing the importance of Rh D alloimmunization profilaxis. PMID- 17420894 TI - [Bioethical study on the expectations of women awaiting assisted reproduction in a public hospital in the Federal District, Brazil]. AB - OBJECTIVE: To analyze the expectations of women who wait for Assisted Reproduction Treatment-RA in the public hospital chosen as the reference in the Public Health Network in the Federal District-HRAS, Brazil. METHODS: For thirty days, 51 women of the 56 who went to the HRAS for infertility treatment were interviewed by a questionnaire including 10 objective questions related to the topic. RESULTS: This trial was divided into two groups. The first, the "control group", comprised 27 patients recently sent to the reference public hospital from local health care centers or a regional hospital. The second, the "study group", comprising 24 women already diagnosed by the medical staff of HRAS and in the waiting line for "in vitro" fertilization. According to the input provided by the two groups, results show that the average waiting time for treatment is so long that women actually age during this time and face the risk of having a dangerous pregnancy before they receive treatment. CONCLUSION: These results show that women unable to pay for treatment in a private fertilization clinic have a poorer chance of achieving RA: the health problem concerning this specific population ignores redressing or income distribution processes. Data show that, notwithstanding, this waiting period imposed by the State, expectations of the patients waiting for RA are reinforced. There is no basis to provide information about the waiting time. The unpredictable availability of the medication needed for in vitro fertilization, jeopardizes the future of this service offering the treatment. PMID- 17420895 TI - [Characteristics related to the first and last cesarean delivery among women from a Campinas University Hospital]. AB - OBJECTIVE: To study the association between first and last caesarian sections with tubal sterilization; to determine length of reproductive life after the first delivery. METHODS: From February to October 2001 in a university hospital, interviews were carried out with 653 women having had at least two pregnancies. Of these women, 172 had a first caesarian section; 294 had a last caesarian section. Variables were social demographic characteristics, obstetric history and characteristics of the first and last deliveries and tubal sterilization. Bivariate analysis was performed, followed by multiple regression analysis calculating the adjusted odds ratio. Women who had undergone tubal sterilization were divided into age groups of 25 to 44 and >45 years in a percentile distribution. The Wilcoxon test was used to analyze age at tubal sterilization and length of reproductive life after the last delivery. The study was approved by the Ethics Committee. RESULTS: Of these women, 89% completed<8 years of school education and 78% were Caucasian. On multiple regression analysis, there was an association between the first and last caesarian section (OR=15.28, 95%CI 8.54 to 27.36), having a partner (OR=3.87, CI95% 1.63 to 9.17) and giving birth in the '70s, '80s or '90s (OR=4.43, 95%CI 1.37 to 14.27), (OR=6.11, 95%CI 1.47 to 25.47) and (OR=6.67, 95%CI 1.21 to 40.26), respectively. The last caesarian section was associated with intrapartum tubal sterilization (OR=14.09, 95%CI 7.37 to 26.97), giving birth in the '70s, '80s or '90s (OR=1.81, 95%CI 1.06 to 3.09), (OR=5.53, 95%CI 3.18 to 9.61) and (OR=5.90, 95%CI 3.03 to 11.48), respectively, family income of >5 minimum wages (OR=2.41, 95%CI 1.42 to 4.08) and age at first delivery>25 years (OR=1.80, 95%CI 1.01 to 3.22). Mean age at sterilization was 29.0 and 33.2 years in women aged 25 to 44 years and >45 years, respectively (p<0.001). The duration of the reproductive period after the first delivery was 9.0 and 11.4 years for the same groups (p<0.001). CONCLUSION: The first caesarian section was associated with the last caesarian section. The last caesarian section was associated with intrapartum tubal sterilization. Age at sterilization was lower and the reproductive period was shorter among younger women. PMID- 17420896 TI - [Tobacco use and plasma lipid-lipoprotein profile in adolescents]. AB - OBJECTIVE: To analyze the impact of tobacco use on plasma lipid lipoprotein profile in representative sample of adolescents. METHODS: A sample of 452 subjects (246 girls and 206 boys) 15 to 18 years old were included in the study. Each participant completed a structured and self-administered questionnaire concerning tobacco use. Plasma lipid-lipoprotein concentrations were measured by standard procedures. Differences between mean values were evaluated by analysis of covariance, controlling for saturated fat and cholesterol intake. Odds ratio was used to estimate the relative risk of the smokers being classified with an undesirable level of a plasma lipidlipoprotein parameter. RESULTS: The proportion of smokers was 20.9% for boys and 15.4% for girls. The average consumption of cigarettes per day was 9.2+/-4.7 for boys and 5.6+/-3.1 for girls. When compared with non-smokers, boy and girl smokers showed a significantly higher serum levels of total cholesterol, LDL-cholesterol, triglycerides and apolipoprotein B100, and significantly lower serum levels of HDL-cholesterol. Adolescent smokers tended to show a two-fold higher risk of altered lipid-lipoprotein levels than non-smokers. CONCLUSION: The present data could imply that intervention promoting a healthy lifestyle, including non smoking, should start at an early age to prevent or delay development of atherosclerotic lesions and ultimately to minimize the appearance of premature coronary heart disease in adults. PMID- 17420897 TI - [Duration of mechanical ventilation and development of bronchopulmonary dysplasia]. AB - OBJECTIVE: Verify the association between duration of mechanical ventilation and development of bronchopulmonary dysplasia in neonates weighting at birth less than 1500 g. METHODS: Retrospective study conducted with neonates weighting less than 1500 g at birth submitted to mechanical ventilation. Neonates presenting major birth defects, transferred to other services or died before the 28th day of life were excluded from the study. Three groups were analyzed according to duration of mechanical ventilation: 1 to 7 days, 8 to 14 days and more than 15 days. The chance ratio of developing bronchopulmonary dysplasia was calculated for each group. RESULTS: From the 216 clinical histories assessed, 121 met the criteria for inclusion in the study. Mean birth weight and gestational age were 1199.8 g and 31.8 weeks. Of all neonates submitted to mechanical ventilation from 1 to 7 days, 15.5% developed bronchopulmonary dysplasia; from 8 to 14 days 60% and from more than 15 days, 88.2%; chance ratios were equal to 0.16; 11.25 and 16.36, respectively. CONCLUSION: The chance of a neonate weighting less than 1500 g developing bronchopulmonary dysplasia was 11 times higher in those submitted to mechanical ventilation for up to 14 days. This chance was even higher in those ventilated for more than 15 days. That is why the nursing staff assisting high risk patients should consider the possibility of extubating neonates during their first week of life. PMID- 17420898 TI - [The physician when facing diagnosis and prognosis of advanced cancer]. AB - Biotechnological scientific progress has brought about some misunderstanding between advances and ethical decisions. The physician-patient relationship when facing seriously ill patients has been altered. OBJECTIVE: To evaluate the physician's attitude when facing disclosure to patient, family or both of cancer diagnosis and prognosis including also a bioethical analysis about the conflicts between beneficence, respect for patient autonomy and paternalism. METHODS: Thirty-eight physicians responsible for neoplasic patients in a tertiary hospital were interviewed. The questionnaire was structured in two different parts. The first, with general information about the physicians and the second, with specific questions about diagnosis and prognosis disclosure to the patient, evaluating also the physician's comprehension of beneficence. RESULTS: Although most physicians (97.4%) inform the diagnosis to their patients in cases of fatal diseases, 50.0% rely on the family for support. In cases of fatal prognosis, 63.1% tell only the families while 31.6% prefer to tell only the patients. The study showed that physicians misunderstand beneficence and paternalism. CONCLUSION: Physicians normally disclose diagnosis to their patients. When they run out of therapeutic resources, their attitudes become conflicting in relation to the principles of beneficence and autonomy. The number of physicians who believe in paternalism as a form of beneficence is still significant. The paternalistic attitude arises especially when therapeutic resources no longer seem to really solve the problem. PMID- 17420899 TI - [Nutritional status and physical performance of elderly in the city of Sao Paulo]. AB - OBJECTIVE: To evaluate the nutritional status (NS) and its association with physical performance tests (PPT) of elderly in the city of Sao Paulo, Brazil. METHODS: Design--Cross-sectional epidemiological study, in a population on a household basis as part of a multicenter study (the SABE Survey), coordinated by the Pan-American Health Organization. Setting-- From January/2000 to March/2001, the SABE survey comprised 2143 elderly (>or=60 yrs old), of both genders, selected by probabilistic sampling. Of these, 1894 (88.4%) took part in this study. The elderly were separated by gender and age group (60-69, 70-79 and>or=80 yrs old) for the tests and the NS was evaluated according to the Body Mass Index (BMI): underweight (BMIor=28 kg/m2). The PPTs included timed, repeated "chair stand" (CS), timed "pick up a pen" (PP) and standing balance (SB). The associations (CS and PP) were verified by the Cox Regression analysis and (SB) by the chi square test. RESULTS: There were statistically significant associations between PP and SB and obese women. CONCLUSION: Results showed association between nutritional status and functional capacity for women and obesity was the nutritional condition associated with the worst performance in the tests. Gender and age group should be considered in the association between nutritional status and physical performance. PMID- 17420900 TI - [Guided healing and skin graft for the treatment of skin wounds]. AB - OBJECTIVE: The present study compared the treatment of skin wounds by means of conducted healing (an innovative method for treatment of secondary healing) and autogenous skin graft. METHODS: Seventeen rabbits were submitted to removal of two skin segments, one on each side of the dorsum. The graft donor area was left as a wound for conducted healing (A) and was submitted only to debridement, local care and dressings. The skin removed from the above mentioned side was implanted as a graft (B) to cover the wound on the other side. Thus, each animal received the two types of treatment on its dorsum (A and B). Rabbits were divided into two groups according to size of the wounds: Group 1--A and B (4 cm2) and Group 2--A and B (25 cm2). Healing time was of 19 days for Group 1 and of 35 days for Group 2. The final macro- and microscopic aspects of healing were comparatively analyzed in the four subgroups. Presence of inflammatory cells, of epidermal cysts and of giant cells was assessed by histology. Data were statistically analyzed by the nonparametric Fisher, Kruskal-Wallis and Wilcoxon tests. RESULTS: No macro- or microscopic difference was observed between conducted healing and skin graft, although conducted healing led to a more rapid wound healing. CONCLUSION: Conducted healing seems to be a good therapeutic option for skin wounds in rabbits. PMID- 17420901 TI - Prevalence of the burnout syndrome among Brazilian medical oncologists. AB - INTRODUCTION: Burnout syndrome which is prevalent among oncologists is characterized by three aspects: emotional exhaustion, depersonalization and low personal accomplishment. The purpose was to evaluate prevalence of the burnout syndrome among Brazilian medical oncologists and the variables that correlate with its presence. METHODS: A survey was conducted with members of the Brazilian Society of Medical Oncology (SBOC) who received three questionnaires (general, Maslach burnout questionnaire and an opinion survey) mailed to all 458 members. RESULTS: Response rate was of 22.3%. According to the criteria proposed by Grunfeld, which consider burnout present when at least one of the aspects is severely abnormal, prevalence of this syndrome was 68.6% (95% confidence interval, CI: 58.68% to 77.45%). By multivariate analysis having a hobby/physical activity, a religious affiliation, older age, living with a companion and rating vacation time as sufficient were correlated significantly and independently with burnout syndrome. CONCLUSIONS: The burnout syndrome is prevalent among Brazilian oncologists. Oncologists having sufficient personal and social resources to engage in a hobby, physical activity, have enough vacation time and religious activities are at lower risk of developing burnout. PMID- 17420902 TI - [Effects of recombinant human erythropoietin in preterm newborns with infectious diseases]. AB - OBJECTIVE: To study the effects of recombinant human erythropoietin (rHuEpo) in preterm newborns (PTNs) with serious infectious diseases. METHODS: A not randomized case-control study was carried out in 34 preterm newborns with diagnosis of serious infectious pathologies, gestational age up to 35 weeks, birth weight less than 1500 g and clinical stability. Newborns selected for treatment with rHuEpo received 400 U/kg erythropoietin ss, subcutaneously twice a week. Oral iron supplementation was initiated when the levels of serum ferritin were lower than 60 mcg/l. The study was continued for six weeks or until the patient was discharged from the hospital. Erythropoiesis, granulopoiesis, thrombocytopoiesis, the need for transfusions and the occurrence of new episodes of infectious disease were analyzed. RESULTS: In the treated group there was a significant increase in the number of reticulocytes, although there was no statistically significant difference between the groups with regard to the number or volume of transfusions. There was no significant difference in neutrophils and platelet values. CONCLUSION: The use of rHuEpo, 800 U/kg/week, in PTNs with infectious diseases was effective in inducing erythropoiesis, without significant changes in the number of neutrophils or platelets. This strategy, and the accurate control of the blood collected for laboratory exams, may be beneficial for prevention of the anemia in PTNs with serious infectious diseases. PMID- 17420903 TI - Leisure-time physical activity during pregnancy in the 2004 Pelotas Birth Cohort Study. AB - OBJECTIVE: Physical activity during pregnancy is a poorly investigated subject on population level. The study aimed to describe duration, type and frequency of leisure-time physical activity during pregnancy, and to explore its associated factors. METHODS: A population-based study was carried out during 2004 in Southern Brazil. A total of 4,471 mothers were interviewed soon after delivery. Physical activity was measured using a questionnaire, developed for the study. Results were obtained by Poisson regression. RESULTS: In the sample, 14.8% of women reported to engage in some type of physical activity prior to pregnancy and 12.9% during pregnancy. In the first trimester, 10.4% of all mothers engaged in some type of physical activity; 8.5% in the second trimester and 6.5% in the third trimester. Only 194 mothers (4.3%) were active during the whole pregnancy. In the adjusted analysis, leisure-time physical activity was positively associated with schooling, physical activity advice during prenatal care, and family income (p<0.001), being employed during pregnancy (p=0.05), and number of pregnancies (p=0.02). Walking was the most frequent activity. CONCLUSIONS: The prevalence of leisure-time physical activity is low among Brazilian pregnant women. Although physical activity is not perceived as being pregnancy threatening, and current guidelines recommend it, this population's behavior does not seem to be changing. Active lifestyle for both pregnant women and future mothers should be encouraged. PMID- 17420904 TI - [II Brazilian guidelines for the management of chronic cough]. PMID- 17420905 TI - [IV Brazilian Guidelines for the management of asthma]. PMID- 17420906 TI - Dengue and laboratory tests a clinician should know for a correct diagnosis. PMID- 17420907 TI - Enrichment methodology to increase the positivity of cultures from body fluids. AB - Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1) the conventional culture method (agar plating) and (2) the enrichment culture technique, using the Bact/Alert blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents. PMID- 17420908 TI - Intestinal permeability and malabsorption of rifampin and isoniazid in active pulmonary tuberculosis. AB - Low antimycobacterial drug concentrations have been observed in tuberculosis (TB) patients under treatment. The lactulose/mannitol urinary excretion test (L/M), normally used to measure intestinal permeability, may be useful to assess drug absorption. The objective of this research was to study intestinal absorptive function and bioavailability of rifampin and isoniazid in TB patients. A cross sectional study was done with 41 patients and 28 healthy controls, using the L/M test. The bioavailabilities of rifampin (R) and isoniazid (H) were evaluated in 18 patients receiving full doses. Urinary excretion of mannitol and lactulose, measured by HPLC, was significantly lower in TB patients. The serum concentrations of the drugs were below the expected range for R (8-24 mcg/mL) or H (3-6 mcg/mL) in 16/18 patients. Analyzing the drugs individually, 12/18 patients had low serum concentrations of R, 13/18 for H and 8/18 for both drugs. We suggest that there is a decrease in the functional absorptive area of the intestine in TB patients, which would explain the reduced serum concentrations of antituberculosis drugs. There is a need for new approaches to improve drug bioavailability in TB patients. PMID- 17420909 TI - Urinary tract infection in non-hospitalized patients with cirrhosis and no symptoms of urinary tract infection: a case series study. AB - Bacterial infections are important factors in decompensation, and they increase the mortality rate of patients with liver cirrhosis. The most common infections among these patients are spontaneous bacterial peritonitis, pneumonia, skin infections and urinary tract infections (UTI). This transversal study evaluated the frequency of UTI in non-hospitalized patients with cirrhosis followed in a hepatology outpatient unit. Patients with clinical, laboratorial, echographic and/or histological diagnosis of cirrhosis were evaluated from April 2002 to August 2004. Patients who accepted participating in this study were submitted to clinical evaluation and the following laboratorial examinations: urine analysis, urine culture, blood culture and hepatic function tests. Patients with symptoms of UTI, diabetis, prostatic disease were excluded. Eighty-two patients with cirrhosis were studied. Their mean age was 51 years (SD = 11); 73% were male. Hepatitis C virus was the main etiology in 45% of the cases. The Child-Pugh B functional class was observed in 52% of the cases. Urine cultures were positive in 4.9% of these patients. In this study of non-hospitalized cirrhotic patients, with no symptoms of UTI, the frequency of urinary tract infection was approximately 5%. The bacteria found were E. coli and Klebsiella pneumonia. We conclude that it is necessary to screen for UTI in such patients. PMID- 17420910 TI - Nosocomial diarrhea in the intensive care unit. AB - We made an epidemiological case-control study to examine risk factors for the development of diarrhea in the intensive care unit (ICU) of a public hospital in Santo Andre, SP, from January to October 2002. Forty-nine patients with diarrhea (cases) and 49 patients without diarrhea (controls), matched for age and gender, were included in the study. A stool culture and enzyme immunoassays for Clostridium difficile toxins A and B were performed on fecal specimens from diarrhea patients. Fourteen of them presented positive cultures for Pseudomonas aeruginosa and 22 patients presented positive ELISA for Clostridium difficile. Nosocomial diarrhea was associated with several factors, including use of antibiotics (P=0.001), use of ceftriaxone (P=0.001), presence of infection (P=0.010) and length of hospital stay (P=0.0001). PMID- 17420911 TI - Occurrence of extended-spectrum beta-lactamases in Enterobacteriaceae isolated from hospitalized patients in Curitiba, southern Brazil. AB - Production of extended-spectrum beta-lactamases (ESBL) by enterobacteria is an important resistance mechanism against antimicrobial beta-lactamics. We tested 498 bacterial strains isolated from two tertiary-care teaching hospitals for ESBL production, using screening breakpoints for aztreonam and third generation cephalosporins, according to CLSI recommendations. Among these isolates, 155 were positive for the ESBL screening test, and 121 (78%) were confirmed by the clavulanic acid combination disk method. We found a high frequency of ESBL (24%) among Enterobacteriaceae, with a frequency of 57.4% for Klebsiella pneumoniae, 21.4% for Klebsiella oxytoca, and 7.2% for E. coli. In other members of Enterobacteriaceae, non-Klebsiella and non-E. coli, the prevalence was 21.6%. Ceftriaxone and cefotaxime showed a higher sensitivity in the screening test (99.2%) when compared to ceftazidime, aztreonam and cefpodoxime. However, cefotaxime/cefotaxime plus clavulanic acid showed a higher sensitivity in the confirmatory test (96.7%). PMID- 17420912 TI - Evaluation of the incidence of occult bacteremia among children with fever of unknown origin. AB - We reviewed the incidence of occult bacteremia, to identify the most frequent etiological agents of bacteremias in otherwise healthy children from one month to 10 years old, who had fever of unknown origin attended at the emergency ward of an urban, university-affiliated pediatric referral center. This was a retrospective medical record review, evaluating children with fever. Data were collected from the initial visit, when blood cultures, hematological properties and hemosedimentation rates were examined. Fever was considered as the highest temperature assessed in the hospital or reported by the responsible adult. Occult bacteremia was discovered in 1.4% of the 1,051 children evaluated, and the most common etiologic agent was Streptococcus pneumoniae. Total leukocyte count and blood sedimentation rates greater than 30 mm(3) were not predictive factors for occult bacteremia. Fever greater than 39 masculineC was the most important factor for predicting occult bacteremia (P<0.001). The presence of occult bacteremia was significantly correlated with patient hospitalization. PMID- 17420913 TI - Antibacterial activity of Brazilian Amazon plant extracts. AB - Infections caused by multiresistant bacteria are a widespread problem, especially in intensive care units. New antibiotics are necessary, and we need to search for alternatives, including natural products. Brazil is one of the hottest spots in the world in terms of biodiversity, but little is known about the chemical and pharmacological properties of most of the plants found in the Amazon rain forest and the Atlantic Forest. We screened 1,220 organic and aqueous extracts, obtained from Amazon and Atlantic rain forest plants, against Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa and E. coli. Seventeen organic and aqueous extracts obtained from 16 plants showed activity against both Gram positive bacteria. None of the extracts showed relevant activity against the Gram negative E. coli and Pseudomonas aeruginosa. PMID- 17420914 TI - Central nervous system involvement in Hodgkin's lymphoma associated with Epstein Barr virus in a patient with AIDS: case report and review of the literature. AB - Intracranial and intraspinal involvement is a rare complication of Hodgkin's disease. We report a case of a patient with diagnosis of AIDS and Hodgkin's lymphoma who developed brain and spinal involvement at the time of the relapse of the neoplasm disease. Mixed cellularity histology was the subtype of Hodgkin's disease in our patient; we identified the Epstein-Barr virus genome in the Reed Sternberg cells by immunohistochemistry and in situ hybridization. PMID- 17420915 TI - Psychosis during peginterferon-alpha 2a and ribavirin therapy: case report. AB - Pegylated Interferon-alpha, combined with ribavirin, gives high sustained virological response in patients with hepatitis C virus, an important public health problem and one of the most frequent chronic infectious diseases worldwide. Though it has therapeutic benefits, treatment with IFN-alpha may be complicated by various side effects, especially symptoms of major depression and acute mania. Psychosis is a rare side effect, and its management usually includes discontinuation of IFN-alpha. We report a case of psychotic disorder that occurred during therapy with pegylated Interferon-alpha given associated with ribavirin. After good response to psychiatric treatment, it became possible to finish the anti-viral therapy. PMID- 17420916 TI - Response of HIV-associated proteinuria to antiretroviral therapy in HIV-1 infected children. AB - HIV-associated nephropathy has been found in children with HIV-1 infection as a late manifestation of this disease; it is associated with nephrotic syndrome with focal segmental glomerulosclerosis and/or mesangial hyperplasia with microcystic tubular dilatation. This is quite rare in children and no cases have been reported from India. Several mechanisms have been hypothesized for the HIV induced renal damage. We report on two HIV-infected children with HIV-associated proteinuria and dramatic response to antiretroviral therapy. PMID- 17420917 TI - Bartonellosis: suggestive case reports in adult and pediatric patients and therapeutic issues. AB - Cat-scratch disease warrants extensive investigation, from an epidemiological, a diagnostic, but especially a therapeutic point of view. Two suggestive episodes of Bartonella henselae-caused cat-scratch disease are reported, and discussed in the light of the most recent literature evidence. The first case occurred in a 60 year-old man, thus suggesting that it is important to maintain an elevated level of suspicion for this disease in adults as well. Both episodes were characterized by a very prolonged and complicated disease course (with the involvement of three lymph node sets in the first case), a need for lymph-node drainage, and apparently negligible activity of many antimicrobial courses, with a very slow local cure. While specific culture and molecular biology techniques proved negative (probably due to late availability of appropriate clinical specimens), indirect immunofluorescence antibody assay was positive since the first weeks of disease, and elevated levels were also fond many months after disease onset. When clinicians face patients with prominent swelling of lymph nodes draining from the upper limbs, cat-scratch disease may be suspected on the grounds of epidemiological and clinical features, with a limited systemic involvement contrasting with a prominent local disease. The significance of specific antibody temporal kinetics in the subacute disease course is still unknown. Although biomolecular assays are now available, the time elapsed from disease onset to clinical diagnosis usually hampers diagnosis, while the roles of surgical debridement and of the unpredictable activity of antimicrobial chemotherapy warrant careful investigation. PMID- 17420918 TI - Fatal case of bacteremia caused by an atypical strain of Corynebacterium mucifaciens. AB - Corynebacterium species have often been considered normal skin flora or contaminants; however, in recent years they have been increasingly implicated in serious infections. Moreover, many new species have been discovered and old species renamed, especially after molecular biology techniques were introduced. Corynebacterium mucifaciens is mainly isolated from blood and from other normally sterile body fluids; it forms slightly yellow, mucoid colonies on blood agar. We report a fatal case of bacteremia due to an atypical strain of C. mucifaciens. This strain had atypical colony morphology; analysis of the 16S rRNA gene was used to define the species. PMID- 17420919 TI - Disseminated fusariosis presenting with pulmonary nodules following a line infection. PMID- 17420920 TI - Purified Protein Derivative--PPD. PMID- 17420921 TI - The role of hereditary spastic paraplegia related genes in multiple sclerosis. A study of disease susceptibility and clinical outcome. AB - Multiple sclerosis (MS) is a common inflammatory disease of the central nervous system unsurpassed for its variability in disease outcome. It has been observed that axonal loss in MS is significant and that irreversible clinical disability relates to such axonal loss. The clinical similarities between Hereditary Spastic Paraplegia (HSP) and progressive MS, along with their analogous profiles of axonal loss in the long tracts, make the genes known to cause HSP biologically relevant candidates for the study of clinical outcome in MS. A cohort of sporadic MS cases and a set of unaffected controls were used to determine the role of HSP genes on MS susceptibility and disease severity. The MS cases were taken from opposite extremes of the putative distribution of long-term outcome using the most stringent clinical criteria to date. Genotyping the two sets of MS patients and controls could not provide any evidence to suggest that genes involved in the pathogenesis of HSP (Paraplegin, NIPA1, KIF5A, HSPD1, Atlastin, Spartin, Spastin, PLP1, L1CAM, Maspardin and BSCL2) play a role in susceptibility to, or modifying the course of, MS, although small effects of these genes cannot be ruled out. PMID- 17420922 TI - Anti-Ma2 antibodies in B-cell primary CNS lymphoma. PMID- 17420923 TI - CSF neurofilament protein (NFL) -- a marker of active HIV-related neurodegeneration. AB - BACKGROUND AND METHODS: The light subunit of the neurofilament protein (NFL), a major structural component of myelinated axons, is a sensitive indicator of axonal injury in the central nervous system (CNS) in a variety of neurodegenerative disorders. Cerebrospinal fluid (CSF) NFL concentrations were measured by ELISA (normal < 250 ng/l) in archived samples from 210 HIV-infected patients not taking antiretroviral treatment: 55 with AIDS dementia complex (ADC), 44 with various CNS opportunistic infections/tumours (CNS OIs), 95 without neurological symptoms or signs, and 16 with primary HIV infection (PHI). The effect of highly active antiretroviral treatment (HAART) was studied by repeated CSF sampling in four of the ADC patients initiating treatment. RESULTS: CSF NFL concentrations were significantly higher in patients with ADC (median 2590 ng/l, IQR 780-7360) and CNS OIs (2315 ng/l, 985-7390 ng/l) than in neuroasymptomatic patients (<250 ng/l, <250-300) or PHI (<250 ng/l, <250-280), p < 0.001. Among patients with ADC, those with more severe disease (stage 2-4) had higher levels than those with milder disease (stage 0.5-1), p < 0.01. CSF NFL declined during HAART to the limit of detection in parallel with virological response and neurological improvement in ADC.CSF NFL concentrations were higher in neuroasymptomatic patients with lower CD4-cell strata than higher, p < 0.001. This increase was less marked than in the ADC patients and noted in 26/58 neuroasymptomatic patients with CD4 counts <200/microl compared to 1/37 with CD4 cells > or =200/microl. CONCLUSIONS: The findings of this study support the value of CSF NFL as a useful marker of ongoing CNS damage in HIV infection. Markedly elevated CSF NFL concentrations in patients without CNS OIs are associated with ADC, follow the grade of severity, and decrease after initiation of effective antiretroviral treatment. Nearly all previously suggested CSF markers of ADC relate to immune activation or HIV viral load that do not directly indicate brain injury. By contrast NFL is a sensitive marker of such injury, and should prove useful in evaluating the presence and activity of ongoing CNS injury in HIV infection. PMID- 17420924 TI - A novel mutation in the HSPD1 gene in a patient with hereditary spastic paraplegia. AB - A mutation in the HSPD1 gene has previously been associated with an autosomal dominant form of spastic paraplegia in a French family. HSPD1 encodes heat shock protein 60, a molecular chaperone involved in folding and quality control of mitochondrial proteins. In the present work we have investigated 23 Danish index patients with hereditary spastic paraplegia (HSP) for mutations in the HSPD1 gene. One patient was found to be heterozygous for a c.1381C > G missense mutation encoding the mutant heat shock protein 60 p.Gln461Glu. The mutation was also present in two unaffected brothers, but absent in 400 unrelated Danish individuals. We found that the function of the p.Gln461Glu heat shock protein 60 was mildly compromised. The c.1381C > G mutation likely represents a novel low penetrance HSP allele. PMID- 17420925 TI - The incidence of motor nueron disease in Scotland. AB - Between 1989 and 1998, 1226 cases of ALS/MND were identified in Scotland, with mean age of onset 65.2 (SD 11.9) years for men and 67.2 (SD 11.0) for women. Annual standardized incidence was 2.40 per 100,000 (95% CI 2.22-2.58). Using capture recapture methods we confirm a high level of case ascertainment for each year of study. Incidence and ascertainment of ALS has remained stable in a large population over a prolonged period of time. Large population-based databases can be used to test aetiological hypotheses. PMID- 17420926 TI - The perception of passive motion in Parkinson's disease. AB - The perception of limb motion is a kinaesthetic property that is essential for voluntary motor control. This study examined the ability of patients with Parkinson's disease (PD) to detect the velocity of a passively moved limb. Eight patients with mild to moderate PD and eight age-matched healthy controls participated. They placed their forearm on a padded splint of a passive motion apparatus, which horizontally extended or flexed the elbow joint at velocities between 1.65 and 0.075 degrees /s (in steps of 0.15 degrees /s). Passive movement persisted until subjects detected arm motion and pressed a trigger held in the hand of their non-tested arm. Time until detection and associated arm displacement were recorded and subsequently adjusted for each subject's reaction time. We found that PD patients needed significantly larger limb displacements before they could judge the presence of passive motion. With decreasing passive motion velocity the detection time increased exponentially in both groups. Yet, the mean detection times of the PD group were 92-166% higher than in the control group for each of the 12 tested velocity conditions. Five of the eight patients were on Parkinsonian medication when tested. Yet, the degree of impairment in the PD group did not correlate significantly with the patients' levodopa equivalent dosage. Our results demonstrate that PD patients were impaired in the detection of passive forearm movements. This study complements a growing body of evidence indicating that various aspects of kinaesthesis (position sense, weight perception, passive motion sense) are affected even at early stages of PD. The impaired processing of proprioceptive signals likely contributes to motor symptoms in PD. PMID- 17420927 TI - Effects of daily water drinking on orthostatic and postprandial hypotension in patients with multiple system atrophy. AB - OBJECTIVE: It has been demonstrated that the increased blood pressure (BP) caused by a single dose of water alleviates orthostatic hypotension (OH) and postprandial hypotension (PPH) in patients with autonomic failure (AF). The aim of this study was to evaluate the practical effect of daily water drinking on OH and PPH in the morning when patients with AF are usually most affected. METHODS: In five patients with multiple system atrophy (MSA) characterized by intractable OH and PPH, we measured seated, standing and postprandial BP in the morning without and with ingestion of 350 ml tap water at 07.30 hours for seven successive days. The changes from the basal BP level at 07.30 hours (DeltaBP) were assessed as an index of the effect of water drinking. RESULTS: Water drinking elicited a rapid pressor response in all patients. The DeltaBP during sitting, standing and after a meal following water drinking (day 1 and day 7) was significantly higher than without water drinking (day 0). The effects of reducing OH and PPH on day 7 were equivalent to those on day 1. No adverse effects associated with daily water drinking were observed, except later diuresis, which occurred in one patient. CONCLUSIONS: Daily water drinking demonstrated constant pressor effects in the morning with no severe adverse effects in MSA patients. This finding suggests that water drinking should be tried as a practical measure to prevent or reduce OH and PPH. PMID- 17420928 TI - Abnormal sudomotor skin responses to temperature and pain stimuli in syringomyelia. AB - Thermoalgesic sensory deficits in patients with syringomyelia may escape objective documentation with conventional electrophysiological techniques. We examined six patients with radiologically proven centrospinal cavities and patchy thermoalgesic sensory deficits by recording the evoked potentials and the sympathetic sudomotor skin responses (SSR) to laser stimuli. While electrical stimuli to the affected areas induced evoked potentials and SSRs of normal latency and amplitude, CO2 laser stimulation induced absent or abnormally reduced evoked potentials. Also, warmth and heat pain stimulation with a Peltier thermode induced absent or abnormal SSRs when applied over the affected areas but well defined SSRs when applied to the corresponding contralateral areas. Our results reveal the utility of recording the SSR to pain and temperature stimuli over specific body sites to demonstrate impairment of pain and temperature pathways in patients with syringomyelia. Comparison of electrical versus laser and temperature induced SSRs is an objective means to evaluate the selective thermoalgesic sensory deficit in these patients. PMID- 17420929 TI - IL2RA/CD25 polymorphisms contribute to multiple sclerosis susceptibility. PMID- 17420930 TI - Pharmacodynamics of interferon beta in multiple sclerosis patients with or without serum neutralizing antibodies. AB - To analyze the in vivo biological effect of anti-interferon beta (IFN-beta) neutralizing antibodies (NABs), blood concentrations of neopterin, beta2microglobulin (Beta2-MG), mRNA-dependent myxovirusresistant protein A (MxA) and dsRNA-dependent protein kinase (PKR) were measured before (predose) and 24 hours after (postdose) IFN-beta administration in 49 patients with multiple sclerosis (MS) with (n = 25) and without (n = 24) NABs. The results indicated that predose levels of MxA-mRNA and PKR-mRNA were highly variable [coefficient of variation (CV) > 100%] among patients. A lower inter-individual variability was observed for pre-dose levels of Beta2-MG and neopterin (CVs of 29% and 44%, respectively). Significantly lower pre- and post-dose blood levels of IFN induced markers, except for postdose PKR-mRNA (p = 0.09), were seen in NAB+ compared with NAB-patients and between patients with high (> 200 t(1/10)) and low ( pound 200 t(1/10)) NAB titers. A significant inverse correlation between NAB titer and pre dose levels of the above IFN-induced markers was found. In summary, our findings confirm that NABs affect absolute concentrations of IFN-beta induced markers and suggest that such an effect occurs in a titer-dependent manner. PMID- 17420931 TI - Recurrent gastrinoma in the mesentery 19 years after primary resection. PMID- 17420932 TI - An unusual case of hemochromatosis in an African-American man: case report and review of the literature. PMID- 17420933 TI - Achalasia in Iceland, 1952-2002: an epidemiologic study. AB - Limited epidemiologic information is available on achalasia in Northern European countries and worldwide. Achalasia has never been studied in Iceland. This study aimed to evaluate the epidemiology of achalasia in Iceland and compare the findings to other studies. All patients diagnosed with achalasia in Iceland from 1952 to 2002 were identified and patients demographic and clinical history was reviewed. The incidence and prevalence of achalasia were obtained. Sixty-two achalasia patients were diagnosed during the 51-year study period, 33 males and 29 females. The mean age at diagnosis was 45.2 years (range, 13.2-85.4 years). The median duration of symptoms before diagnosis was 2.0 years. The mean prevalence was 8.7 cases/100,000 and the overall incidence was 0.55 case/100,000/year. This is the first national epidemiological study of achalasia in a genetically homogeneous population, spanning over half a century. The epidemiology of achalasia in Iceland is similar to that in most other reported studies. PMID- 17420934 TI - Colonization and impact of disease and other factors on intestinal microbiota. AB - The aim of this study was to review the process of microbial colonization and the environmental and host factors that influence colonization and microbial succession. The impact of some diseases on intestinal microbiota composition is also described. Microbial colonization of the gut by maternal vaginal and fecal bacteria begins during and after birth. During the first 2 years of life, specific microbes become established in a process designated microbial succession. Microbial succession in the gastrointestinal tract is influenced by numerous external and internal host-related factors, and by the second year of life, the intestinal microbiota composition is considered identical to that of adults. Nevertheless, intestinal microbiota in both infants and adults remain incompletely characterized and their diversity poorly defined. The main explanation is that many intestinal bacteria that live in an anaerobic environment are difficult or impossible to culture outside the intestine. However, recent advances in molecular biology techniques have initiated the description of new bacteria species. The composition of gut microbiota can be modulated by host, environmental, and bacterial factors, and strong evidence has emerged of substantial modifications during illness or exposure to threatening experiences. It has been postulated that improvements in hygienic measures have led to an increase in allergic diseases ("hygiene hypothesis"). Alterations in gut microbiota and their functions have been widely associated with many chronic and degenerative diseases, including inflammatory bowel disease, colon cancer, and rheumatoid arthritis. PMID- 17420935 TI - Total colectomy for all colorectal carcinoma cases or follow-up indefinitely? PMID- 17420936 TI - Spontaneous intestinal perforation due to non-Hodgkin's lymphoma: evaluation of eight cases. AB - Primary intestinal lymphoma with spontaneous perforation and after systemic chemotherapy is rare. The present study summarizes retrospectively the outcome of eight free intestinal perforated patients diagnosed with intestinal non-Hodgkin's lymphoma. Two patients had a history of systemic chemotherapy before perforation. The most common symptoms of the patients were abdominal pain, nausea, vomiting, weight loss, and fever. Sites of perforation were ileum in four, jejunum in two, cecum in one, and sigmoid colon in one patient. Synchronous lymphoma was present in three patients. The perforation was closed by primary closure in three patients. Resection/anastomosis was performed in four patients and sigmoid colostomy was performed in one patient. Three patients were lost due to leakage or septicemia. Intestinal lymphoma might be kept in mind as a cause in free intestinal perforations. Because of the high mortality rate early diagnosis and treatment are important to improve the prognosis of bowel perforation in patients with non-Hodgkin's lymphoma. PMID- 17420937 TI - Intragastric nitroglycerin at a vasodilatory dose attenuates acidified aspirin induced gastric mucosal injury. AB - Clinical studies reveal that aspirin intake to prevent myocardial and cerebral ischemia is associated with a significant increase in upper gastrointestinal hemorrhage requiring hospitalization and that nitroglycerin or long-acting nitrates significantly lower this risk. Nitroglycerin can increase gastric blood flow and slow gastric emptying. We hypothesized that both features contribute to its gastroprotective property. Fasted anesthetized rats (Study 1) and conscious mice (Studies 2 to 4) received intragastric nitroglycerin or vehicle. The effects of these two treatments on various parameters were assessed in Study 1, on blood pressure and gastric blood flow; Study 2, on acidified aspirin-induced gastric mucosal lesions; and Study 3, on the weight of gastric content. In Study 4, the effect of nitroglycerin, vehicle, or vehicle plus saline, on acidified aspirin induced gastric mucosal lesion was assessed. Compared with vehicle, nitroglycerin decreased blood pressure and produced a mild but significant increase in gastric vascular conductance, blood flow, and volume of gastric content. The number and length of gastric mucosal lesions induced by acidified aspirin were significantly attenuated by intragastric nitroglycerin in a vasodilatory dose. Exogenous saline in a volume equivalent to the increase produced by nitroglycerin, however, did not attenuate the lesions. These experimental data are consistent with the clinical observation that nitrates lower the risk of aspirin-induced gastrointestinal complications. Confirmation of the efficacy of nitroglycerin and nitrates in preventing such aspirin-induced complications in controlled trials is worthy of consideration by clinical investigators. PMID- 17420938 TI - Expression of apoptotic epithelial cells in biopsy specimens of patients with colitis. AB - We postulated that nuclear dust within the lamina propria beneath the basement membrane of the epithelium in colonic mucosal biopsies of patients with colitis is a form of apoptotic epithelial cells and that its expression correlates with clinical severity. Our aim was to determine the origin of nuclear dust and to explore the correlation between nuclear dust expression and clinicopathologic parameters of colitis. we examined 228 specimens with colitis and 18 normal specimens. The expression rates of nuclear dust were 11.1% (2/18) and 83.8% (191/228) in normal colonic mucosa and colitis, respectively. Cells showing double positive staining with cytokeratin and TdT-mediated uUTP-biotin nick-end labeling technique were apoptotic cells derived from epithelial cells. Nuclear dust expression correlated significantly with inflammation, eosinophil infiltration, edema, and congestion. Our results suggest that interventions directed toward the apoptotic process may be beneficial in the treatment of colitis. PMID- 17420939 TI - Deficiency of invariant NK T cells in Crohn's disease and ulcerative colitis. AB - The aim of this study was to investigate whether immunoregulatory invariant NK T cells are deficient in Crohn's disease or ulcerative colitis. Blood was collected for flow cytometry from 106 Crohn's disease, 91 ulcerative colitis, and 155 control subjects. Invariant NK T cells were assessed by Valpha24 and (alpha galactosylceramide/CD1d tetramer markers. Intracellular cytokine was measured after in vitro anti-CD3 antibody stimulation. Valpha24+ T cells were quantified in ileocolonic biopsies as mRNA by real-time PCR and by immunofluorescence. Circulating invariant NK T cells were 5.3% of the control levels in Crohn's (P < 0.001) and 7.9% of the control levels in ulcerative colitis (P < 0.001). Interleukin-4 production was impaired in Crohn's disease and ulcerative colitis. Intestinal Valpha24 mRNA expression was 7% in Crohn's disease (P < 0.05) and 9% in ulcerative colitis (P < 0.05). Intestinal Valpha24+ T cells were 23% in Crohn's disease but not reduced in ulcerative colitis. We conclude that invariant NK T cells are deficient in Crohn's disease and in ulcerative colitis. PMID- 17420940 TI - Protective effect of soy isoflavones and activity levels of plasma paraoxonase and arylesterase in the experimental nonalcoholic steatohepatitis model. AB - Nonalcoholic steatohepatitis (NASH) is characterized by diffuse fatty infiltration in the liver and ballooning degeneration and inflammation in hepatocytes. We aimed to study the protective effect of soy isoflavones on experimental NASH and their effects on plasma paraoxanese and arylesterase levels in rats. Twenty-eight male rats were divided into four groups: Group 1 (n=7) received an isocaloric normal diet for 8 weeks, Group 2 (n=7) was fed an isocaloric basal diet plus oral soy isoflavone for 8 weeks (100 mg/kg in diet), Group 3 (n=7) received a special diet that was methionine and choline deficient (MCD) and rich in fat for 8 weeks, and Group 4 (n=7) was fed a special diet that was MCD and rich in fat plus oral soy isoflavone for 8 weeks (100 mg/kg in diet). Blood samples were collected to measure plasma malondialdehyde (MDA), paraoxanese, and arylesterase and biochemical parameters. Tissue samples were duly taken for histopathological examination and measurement of tissue MDA levels. Plasma MDA levels were higher in Group 3 than in Groups 1, 2, and 4 (P <0.01, P <0.05, and P <0.05 respectively). Liver tissue MDA levels were also significantly higher in Group 3 compared to Groups 1, 2, and 4 (P <0.001, P <0.001, and P <0.05 respectively). A significant decrease was found in the plasma and liver tissue MDA levels in Group 4 compared to Group 3 (P <0.05 and P <0.05, respectively). The activity levels of plasma paraoxanase and arylesterase were significantly higher in Group 2 than in Groups 1 and 3 (P <0.05 and P <0.01, respectively). Also, the plasma paraoxanase and arylesterase levels were significantly higher in Group 4 compared to Groups 1 and 3 (P <0.05 and P <0.01, respectively). A significant reduction was observed in Group 4 in steatosis, inflammation, necrosis, and fibrosis compared to Group 3 (P <0.05 for each). We conclude that soy isoflavones seem to be effective in preventing liver damage by decreasing lipid peroxidation in the NASH model induced by a MCD diet. They stimulate and increase the activity of the antioxidative paraoxanase enyzme while decreasing the total cholesterol and triglyceride levels. PMID- 17420941 TI - Clinical study on gastrointestinal stromal tumors (GIST) in Iceland, 1990-2003. AB - This is a whole population-based study on clinical symptoms, surgical treatment, and outcome of GIST. All mesenchymal tumors in the digestive tract diagnosed from 1990 to 2003 were identified. All reports were reviewed, all tumors were stained with antibodies to c-kit, and the diagnosis of GIST was confirmed. Clinical, pathological, treatment, and outcome data were analyzed. The study included 53 patients with GIST. The mean age at diagnosis was 65.8+/-13.6 years (SD). Tumor distribution included 62% in the upper, 32% in the middle, and 6% in the lower digestive tract. Mean tumor size was 4.9+/-4.4 cm (SD). Gastrointestinal (GI) bleeding was the main symptom in 53% (20/38) of symptomatic cases; most presented with acute gastrointestinal bleeding. Complete surgical resection was performed in 87% (46/53) of patients. Eight of the 53 tumors (15.1%) metastasized, 7 of which were nongastric. The disease-specific death rate at 5 years was 85%, and 5 year survival after complete resection was 64.1%. We conclude that GISTs are often found incidentally but GI bleeding is the most common presentation. Five year survival is better than previously reported and gastric GIST seems to be more benign than nongastric. GIST seems to metastasize mainly intra-abdominally. PMID- 17420942 TI - A phase I pilot study of autologous heat shock protein vaccine HSPPC-96 in patients with resected pancreatic adenocarcinoma. AB - We performed a phase I pilot study to determine if autologous vaccine HSPPC-96 (gp96, Oncophage) could be purified from completely resected pancreas adenocarcinomas, to determine patient tolerance of vaccine and to explore immune responses and clinical outcomes of these patients. Subjects were vaccinated with 5 microg of autologous HSPPC-96 weekly for 4 doses. Serial ELISPOT assays of T cells for antitumor reactivity were performed. Subjects received neither adjuvant chemotherapy nor radiation. Ten patients received a full course of vaccinations. No dose-limiting toxicities were encountered. Immediate freezing in liquid nitrogen of the tumor specimen resulted in improved vaccine yield. Median overall survival is 2.2 years (Kaplan-Meier estimate). Autologous anti-HSPPC-96 ELISPOT reactivity increased significantly in 1 of 5 patients examined and a second had an increase of unclear significance. Three of 10 treated patients are alive without disease at 2.6, 2.7, and 5.0 years follow-up. There was no observed correlation between immune response and prognosis. This study demonstrates the feasibility of preparing HSPPC-96 from pancreatic adenocarcinomas. Examination of this novel approach using multiple dose levels is 1 approach to further investigate the immunogenicity and clinical utility of HSPPC-96 vaccination in this setting. PMID- 17420943 TI - Comparison of clinical features and patient background in functional dyspepsia and peptic ulcer. AB - To elucidate the clinical features of functional dyspepsia (FD), patients with FD were compared with patients with peptic ulcer. Fifty-eight FD and fifty-nine peptic ulcer patients were compared with respect to clinical features and patient background. In the FD group, symptoms of dyspepsia, especially upper abdominal fullness and nausea, were more common than in the peptic ulcer group. The FD group complained greater distress (severity of the most distressing symptom; P < .001) and showed higher State-Trait Anxiety Inventory (STAI) scores (trait anxiety score; P < .05). A higher proportion of FD patients had consulted another physician (P < .01). Even when subjects from the FD and peptic ulcer group in this study were matched for age and gender and compared with respect to these variables, almost the same characteristics were seen. These results indicate that FD markedly decreases quality of life in a variety of aspects. PMID- 17420944 TI - Applying data mining techniques in the development of a diagnostics questionnaire for GERD. AB - Gastroesophageal reflux disease (GERD) is a common condition, managed mostly in primary care practice. Heartburn and acid regurgitation are considered primary symptoms, and are usually highly specific. However, the symptom spectrum is much wider and in many cases it is difficult to determine whether the patient has GERD or dyspepsia from another origin. The aim of this study is to develop a symptom score and rule for the diagnosis of GERD, using data mining techniques, to provide a clinical diagnostic tool for primary care practitioners in the evaluation and management of upper gastrointestinal symptoms. A diagnostic symptom questionnaire consisting of 15 items and based on the current literature was designed to measure the presence and severity of reflux and dyspepsia symptoms using a 5-point Likert-type scale. A total of 132 subjects with uninvestigated upper abdominal symptoms were prospectively recruited for symptom evaluation. All patients were interviewed and examined, underwent upper gastrointestinal endoscopy, and completed the questionnaire. Based on endoscopic findings as well as the medical interview, the subjects were classified as having reflux disease (GERD) or non-reflux disease (non-GERD). Data mining models and algorithms (neural networks, decision trees, and logistic regression) were used to build a short and simple new discriminative questionnaire. The most relevant variables discriminating GERD from non-GERD patients were heartburn, regurgitation, clinical response to antacids, sour taste, and aggravation of symptoms after a heavy meal. The sensitivity and specificity of the new symptom score were 70%-75% and 63%-78%, respectively. The area under the ROC curve for logistic regression and neural networks were 0.783 and 0.787, respectively. We present a new validated discriminative GERD questionnaire using data mining techniques. The questionnaire is useful, friendly, and short, and therefore can be easily applied in clinical practice for choosing the appropriate diagnostic workup for patients with upper gastrointestinal complaints. PMID- 17420945 TI - Hepatic abscess that formed secondary to fish bone and had a fistula with the ascending colon. AB - We present the case of a liver abscess that formed secondary to foreign bodies and formed a fistula the ascending colon and was successfully treated with percutaneous abscess drainage and colonoscopic removal of foreign bodies. A 64 year-old man presented with right upper and lower quadrant pain of 2 weeks' duration. Abdominal computed tomography was performed, demonstrating a single 3.5 x 1.9-cm abscess of the liver's right lobe and eccentric thickening of the colon wall at hepatic flexure. A percutaneous hepatic drainage catheter was placed under ultrasound guidance. Colonoscopic examination revealed multiple diverticula of the ascending colon and two 1.5-cm long fish bones at the ascending colon near the hepatic flexure. One end of each fish bone had impacted the edematous colonic mucosa and was surrounded by exudate polypoid inflammatory tissue. The fish bones were extracted with forceps. The patient was feeling well and was discharged after 12 days of treatment. PMID- 17420946 TI - Nimesulide inhibits crypt epithelial cell proliferation at 6 hours in the small intestine in CD-1 mice. AB - To determine whether the gut-sparing selectivity of cyclooxygenase-2 inhibitors is related to early crypt kinetic mechanisms, this study compared the primary effects on small intestinal mucosal epithelial cell proliferation and morphometry of a nonselective dual cyclooxygenase inhibitor, indomethacin, with a cyclooxygenase-2 selective inhibitor, nimesulide. Indomethacin downregulated the crypt cell production rate in the proximal small intestine, and nimesulide reduced cell proliferation in the proximal and distal small intestine. Compared to controls, there were smaller proliferating compartments in the crypts in midintestinal segments in both indomethacin- and nimesulide-treated groups, but more dividing cells in the distal intestine in indomethacin-treated group. Crypt cellularity, numbers, and width were unchanged from control values in both treated groups, suggesting a reduction in crypt cell emigration. Despite its selectivity for inhibiting cyclooxygenase-2, nimesulide induces similar but widespread initial effects on intestinal cell kinetics when compared to indomethacin. PMID- 17420947 TI - Gastrojejunostomy and duodenojejunostomy for megaduodenum in systemic sclerosis sine scleroderma: report of a case. PMID- 17420949 TI - Does small bowel intussusception in adults always require surgery? PMID- 17420948 TI - Review article: multiple myeloma and inflammatory bowel disease. AB - Since 1964 only nine cases of multiple myeloma occurring in the setting of inflammatory bowel disease have been reported. Although this occurrence may be a mere unfortunate coincidence, there are sound pathophysiological reasons for such an event. The possibility that chronic inflammatory conditions, immunomodulator therapy, and infliximab can predispose to multiple myeloma and lymphoma is reviewed. We discuss in detail the only reported case of multiple myeloma arising in the setting of infliximab treatment for Crohn's disease. It is highly probable that infliximab therapy had a causal role in our patient developing multiple myeloma. The pathogenesis of multiple myeloma arising in the setting of infliximab therapy may be related to decreased apoptosis of plasma cell populations. Since it is possible that a causal association exists between infliximab therapy and multiple myeloma, additional screening measures may be required in patients with Crohn's disease on infliximab. PMID- 17420950 TI - Prognostic index for survival in patients after treatment for primary hepatocellular carcinoma. AB - We retrospectively evaluated clinical factors affecting long-term survival after treatment for hepatocellular carcinoma (HCC) to predict the reliability of the prognosis of patients with HCC. We analyzed 157 patients who received treatment for HCC. The prognostic index (PI) was evaluated using the Cox regression model. The overall cumulative survival rates were 79.7% at 1 year, 51.1% at 3 years, and 24.9% at 5 years. The PI was calculated by the following formula, consisting of five factors: PI = 0.637 x number of tumor lesions + 0.103 x tumor diameter + 1.003 x ascites + 0.915 x portal vein tumor thrombosis - 0.006 x cholinesterase + 2.0. It was found that liver function and progression of the tumor are the most important factors for prognosis after treatment for HCC. The PI, based on five factors, will in future be an appropriate index to predict the prognosis of patients with HCC. PMID- 17420951 TI - The clinicopathological profile of Indian patients with nonalcoholic fatty liver disease (NAFLD) is different from that in the West. AB - There are limited data on nonalcoholic fatty liver disease (NAFLD) from India. The clinicopathological profile of Indian patients with NAFLD may be different from that of Western patients. One hundred NAFLD patients with increased liver enzymes were prospectively evaluated for clinical presentation, associated diseases, overweight/obesity, central obesity (n=54), presence of diabetes mellitus, lipid abnormalities, insulin resistance (n=39), metabolic syndrome (n=54), serum iron, serum ferritin, and transferrin saturation (n=60), and HFE gene mutations (n=30). Risk factors for the grade and stage of the disease on histology were studied in 38 biopsy-proven patients. Patients were treated with lifestyle modifications and ursodeoxycholic acid (UDCA). Seventeen nonresponder patients were treated with metformin. The majority of patients were males (n=70). Twenty percent of patients were overweight, 68% had obesity, and 78% had central obesity. Abnormal cholesterol, HDL, and triglycerides were present in 36%, 66%, and 53% of patients, respectively. Twelve percent of patients had diabetes mellitus and 16% patients had various associated diseases. All 22 (100%) patients studied by ITT and all but 1 (98%) studied by HOMA-IR were found to have reduced insulin sensitivity and 50% were found to have metabolic syndrome by the modified ATP III criteria. Two (3%) patients were found to have high serum iron, 4 (7%) patients had high ferritin, 5 (8%) patients had increased transferrin saturation, and 4 (13%) patients were found to be heterozygotes for H63D HFE gene mutation. Twenty patients of 38 (53%) had histological evidence of NASH (class 3=6, class 4=14). The other 18 (47%) qualified for class I (n=1) or class II (n=17) NAFLD. Four (10.5%) patients had bridging fibrosis and none had evidence of cirrhosis liver. Seventy-four (74%) patients achieved a biochemical response to lifestyle modification and UDCA. All 17 patients treated with metformin had a reduction in ALT level and 10 (59%) of them had normalization of their enzymes. We conclude that the clinicopathological profile of NAFLD in Indian patients is different from that in the West. PMID- 17420952 TI - Effect of 5-HT(2) receptor blockade on cadmium-induced acute toxicity. AB - The protective effect of 5-HT(2) receptor blockade with ketanserin or ritanserin against cadmium liver injury was investigated. Male Wistar rats were injected intraperitoneally with a sublethal dose of cadmium (3.5 mg/kg body weight). Rats were treated with normal saline (group I), ketanserin (3 mg/kg body weight; group II), or ritanserin (3 mg/kg body weight; group III) 2 hr prior and 4 hr after cadmium administration and killed at different time points. Hematoxylin/eosin stained liver sections were assessed for necrosis, apoptosis, peliosis, mitoses, and inflammatory infiltration. Apoptosis was also quantified by the TUNEL assay. Nonparenchymal liver cells and activated Kupffer cells were identified histochemically. Necrosis, hepatocyte apoptosis, nonparenchymal cell apoptosis, and macroscopic and microscopic peliosis were markedly reduced or minimized in ketanserin- or ritanserin-treated rats. The observed protective effect was almost identical for both ketanserin and ritanserin administration. 5-HT(2) receptor blockade exerts a protective effect against acute cadmium-induced hepatotoxicity. PMID- 17420953 TI - Lactobacillus plantarum 299v enhances the concentrations of fecal short-chain fatty acids in patients with recurrent clostridium difficile-associated diarrhea. AB - Our objective was to document how intake of Lactobacillus plantarum 299v affects the concentrations of fecal organic acids during and after metronidazole treatment in 19 patients with recurrent Clostridium difficile-associated diarrhea. Fecal samples were analyzed by gas-liquid chromatography. After intake of metronidazole a significant decrease in total short-chain fatty acids was seen in the placebo group (from 77.1 to 45.5 micromol/g; P=0.028) but not in the Lactobacillus group (79.8-60.4 micromol/g). In addition, a statistically significant difference between treatment groups was noted for butyrate (5.6-1.2 micromol/g in the placebo group vs. 7.6-5.6 micromol/g in the Lactobacillus group; P=0.047). At the end of the study and after cessation of placebo or Lactobacillus, the total short-chain fatty acids rose to the same levels as before antibiotic treatment in the placebo group. Both treatment groups showed a significant decrease in concentrations of succinate at the end of the study in comparison to the time when metronidazole intake was stopped (6.3-1.5 micromol/g in the placebo group versus 9.3-0.9 micromol/g in the Lactobacillus group; P=0.028). The present study of fecal samples from a clinical trial is the first to demonstrate that administration of Lactobacillus plantarum 299v reduces the negative effects of an antibiotic on colonic fermentation. The intake of this probiotic strain may thereby provide an additional benefit for patients with recurrent Clostridium difficile-associated diarrhea. PMID- 17420954 TI - Utility of capsule endoscopy for the classification of Crohn's disease: a multicenter study in Korea. AB - Capsule endoscopy can detect lesions associated with Crohn's disease in the small bowel that go unrecognized by conventional radiologic imaging. The aim of this study was to assess the value of capsule endoscopy in the classification of patients with Crohn's disease. This study was carried out on 52 patients with known Crohn's disease. Small bowel follow-through findings were compared with capsule endoscopic findings. All patients were classified according to the extent of the disease: small bowel-colon type, small bowel type, or colon type. Seventeen patients who were reported as being normal showed evidence of small bowel Crohn's disease according to capsule endoscopy. Differences were observed between the small bowel follow-through and the capsule endoscopy classification (kappa=0.54). Treatment was changed based on the results of wireless capsule endoscopy in 15 patients. PMID- 17420955 TI - The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene promoter through binding to E- and G-box elements. AB - Microsomal oleic acid desaturase (FAD2) catalyzes the first extra-plastidial desaturation in plants, converting oleic acid to linoleic acid, which is a major constituent in all cellular membranes as well as in seed storage oils. Seed specific FAD2 (SeFAD2) produced 40% of linoleic acids in the total fatty acids of sesame (Sesamum indicum) seeds. The expression of SeFAD2 transcripts was spatially and temporally controlled during seed development. To investigate the regulatory mechanism controlling seed-specific SeFAD2 expression, we isolated a well-matched sequence homologous to the basic region/helix-loop-helix proteins by yeast one-hybrid screening and named it SebHLH. SebHLH transcripts were expressed in developing seeds and roots of sesame. SebHLH:GFP fusion protein localized in the nucleus. Recombinant SebHLH protein bound E-box (CANNTG) and G-box (CACGTG) elements in the region from -179 to -53 of the SeFAD2 gene promoter, and the external C and G nucleotides in the E- and G-box motifs were essential for SebHLH protein binding. The SebHLH gene, under the CaMV35S promoter, and the GUS reporter gene driven by E- and G-box motifs were co-expressed in developing sesame seeds and Arabidopsis transgenic leaves. This co-expression demonstrated that SebHLH protein mediates transactivation of the SeFAD2 gene promoter through binding to E- and G-box elements. E- or G-box elements frequently occur in the 5' flanking region of genes that are involved in triacylglycerol biosynthesis and that exhibit seed-specific expression in Arabidopsis and other plants, suggesting that bHLH transcription factors play a key role in the transcriptional regulation of genes related to storage lipid biosynthesis and accumulation during seed development. PMID- 17420956 TI - Strand compositional asymmetries in vertebrate large genes. AB - Both transcription-associated and replication-associated strand compositional asymmetries have recently been shown in vertebrate genomes. In this paper, we illustrate that transcription-associated strand compositional asymmetries and replication-associated ones coexist in most vertebrate large genes, although in most case the former conceals the latter. Furthermore, we found that the transcription-associated strand compositional asymmetries of housekeeping genes are stronger than those of somatic cell expressed genes. Together with other evidence, we suggest that germline transcription-associated strand asymmetric mutations may be the main cause of the transcription-associated strand compositional asymmetries. PMID- 17420957 TI - Nodular endometriosis: dynamic MR imaging. AB - PURPOSE: In this study we aimed to investigate the value of contrast enhanced dynamic MR imaging (DMI) in the diagnosis of nodular abdominal endometriosis. SUBJECTS AND METHODS: Fourteen patients with surgically and pathologically proven endometriosis were examined with DMI. The patients were 22-54 years old (mean age 30.8 years). The dynamic MR studies of these patients were retrospectively reviewed by two radiologists who were aware of the clinical data. Nodular masses showing enhancement were evaluated for size, margins, and signal intensity on T1- and T2-weighted MR sequences. The protocol was tailored to selectively determine the diagnostic utility of signal intensity time course analysis for the behavior of nodular endometriosis and endometrial tissue, in DMI. Contrast-enhanced DMI was performed and the time-intensity curves of the lesions and the uterine endometrial tissue of each patient were compared. Mean enhancement values were calculated. Each DMI was evaluated for signal intensity value. RESULTS: In 8 (57%) of 14 patients, we found endometriosis in the abdominal wall. All patients with abdominal wall endometriosis had pelvic surgical operation history. Diameter of nodular endometriosis determined in the abdominus muscle ranged between 3 and 40 mm. Of eight cases, five had only one lesion and three had multiple lesions. Remaining 6 (43%) cases had deep pelvic endometriosis located in the uterosacral ligaments (n = 3), rectosigmoid (n = 2), and rectovaginal septum (n = 1). Diameter of pelvic endometriosis ranged between 9 and 53 mm. Noncontrast mean signal intensity of endometriosis and endometrial tissue were 280 +/- 73 and 216 +/- 20, respectively. The mean values of both endometriosis and normal endometrial tissue were calculated for each patient examined with five-slice DMI. All of the curves showed significant correlation. The lesion showed significant enhancement in the course of time similar to the endometrial tissue in all patients. CONCLUSION: Our study was inspired from the fact that endometriosis is the ectopic endometrial tissue and we thought that endometrial tissue and endometriomas should have similar vascularity. In this way imaging with MR, getting the time-intensity curves and experiencing the correlation between the endometriosis and endometrial tissue may support the diagnosis in the cases with suspected endometriosis. This first study shows that the ectopic nodular endometriosis can easily be identified with dynamic MRI. It may be used to differentiate nodular endometriosis from the other pathologic conditions of abdominal wall and pelvis. PMID- 17420958 TI - Contrast-enhanced sonography of the kidney. AB - Contrast-enhanced sonography (CEUS) is a recently introduced, promising technique in the evaluation of the kidney. CEUS allows real-time assessment of normal and abnormal renal perfusions. As a consequence of the macrocirculation analysis allowed by Doppler techniques, it is possible to obtain real-time information about microcirculation. US contrast media are not nephrotoxic and can be employed safely, even in subjects with impaired renal function. There are several clinical scenarios where CEUS may play the role of a low-cost, scarcely invasive tool, including renal tumors (with special reference to small, indeterminate masses, i.e., differentiation between carcinoma and angiomyolipoma), renal atypical cystic masses (i.e., differentiation of malignant from benign cysts and follow-up of cystic lesions managed conservatively), renal infarction, renal infections, and renal injuries. In addition, CEUS can be useful in the assessment of renal pseudotumors (including any case with possible renal mass on conventional US imaging) and has been employed in radiofrequency ablation guidance. This pictorial review illustrates the CEUS findings recognizable in a wide spectrum of renal disorders and discusses the strengths and limitations of renal imaging with CEUS. PMID- 17420959 TI - Neurectomy to prevent persistent pain after inguinal herniorraphy: a prospective study using objective criteria to assess pain. AB - BACKGROUND: Although tension-free mesh repair has markedly improved the outcome of inguinal hernia surgery, it has only minimally reduced the incidence of persistent postoperative pain. The pathogenesis of this complication and treatment remain unclear. STUDY DESIGN: In order to objectively assess whether iliohypogastric neurectomy reduces the incidence and intensity of persistent postoperative pain, we prospectively studied 100 male patients with bilateral inguinal hernia who underwent tension-free surgical repair, combined with iliohypogastric neurectomy on the right side alone. Pain was evaluated postoperatively on days 1 and 7 and at 1 and 2 years by means of a visual analog scale. Patients were given a questionnaire including coded terms for describing pain. These terms were designed to compare pain on the neurectomized and non neurectomized sides and assess altered sensation (hypoesthesia and paresthesia) on both sides. RESULTS: There were differences in the incidence and intensity of pain between the neurectomized and non-neurectomized sides, though these differences were not significant. Individual patient assessment showed that from postoperative day 7 onward patients had on average less pain on the neurectomized side. Pain reduction was more prominent in patients who scored 4 or more on the visual analog scale. No significant difference was found in the incidence of sensory alterations between the two sides. Two years after inguinal hernia repair, only one of the 100 patients studied still had persistent pain (>4 on the visual analog scale); this pain was on the non-neurectomized side. CONCLUSIONS: Our prospective data do not reach statistical significance to claim that iliohypogastric neurectomy reduces the incidence and intensity of persistent postoperative pain after tension-free inguinal hernioplasty. Studies on larger patient samples are warranted to provide definitive, statistically supported conclusions. PMID- 17420960 TI - Survival after parathyroidectomy in patients with end-stage renal disease and severe hyperparathyroidism. AB - BACKGROUND: Patients with end-stage renal disease (ESRD) and secondary hyperparathyroidism (SHPT) are at high risk of mortality. Whether an increased risk of death persists after a parathyroidectomy (PTX) is not clearly established. SUBJECTS AND METHODS: The survival of 40 patients with ESRD and SHPT who underwent PTX was compared with that of 664 ESRD patients. RESULTS: From first dialysis, a lower mortality rate was found in the group of patients who underwent PTX than in the nonoperated ESRD group (hazard ratio: 0.23; 95% CI: 0.14-0.37). The patients who underwent PTX were younger, had a longer time on dialysis, and had a higher prevalence of kidney transplantation. The mean number of comorbidities was lower (Charlson score 4.2 +/- 2.1 versus 6.4 +/- 2.9, p < 0.001). Then, we randomly selected two matched controls for each PTX case (80 controls, 40 PTX) who had at least an equivalent mean duration of dialysis between the first dialysis and PTX of the PTX group. In a univariate model, there was a trend for PTX being associated with prolonged survival. The mortality was higher both among those at an advanced age and those with a high Charlson score. Adjustments for these covariates made the effect of PTX no more significant. CONCLUSIONS: The risk of death of patients with severe SHPT leading to PTX differed from that of nonoperated subjects. The apparent differences in survival may be related to the number and severity of associated comorbidities. ESRD patients who undergo PTX may represent a subset of healthier subjects. PMID- 17420961 TI - Small-bowel obstruction secondary to bezoar impaction: a diagnostic dilemma. AB - BACKGROUND: Gastrointestinal bezoar (GIB) is uncommon and is reported to occur in 4% of all admissions for small-bowel obstruction (SBO). Because of a lack of diagnostic features, it is often associated with a delay in treatment, with increased morbidity. In this article, we report our experience with managing bezoar-induced SBO and the role of early computed tomography (CT) imaging in establishing the diagnosis. METHODS: We retrospectively reviewed all cases of bezoar-induced SBO treated in our unit between 1999 and 2005. RESULTS: There were 43 patients, of whom 2 had a recurrence, giving a total of 45 episodes. The frequency of bezoar in our patients presenting with SBO was 4.3%. All patients were of Asian origin: 41 Chinese, 1 Indian, and 1 Malay. Twenty-eight (65%) patients had previous abdominal surgery of which 26 were gastric surgery. Thirty eight (88%) patients were edentulous. Forty-one (91%) underwent serial abdominal radiography, whereas only 4 patients (9%) had either CT imaging or contrast study alone. Only 11 (24%) cases had a correct diagnosis of bezoar impaction made preoperatively by CT imaging. The diagnostic accuracy of CT imaging in our series was 65%, with six cases of misdiagnosis. Overall, CT led to a change in management of 76% (13 in 17). The median time to surgery from admission was 2 (0 10) days. There were 2 cases of ischemic bowel that necessitated bowel resection. The median length of hospital stay was 11 (5-100) days. Ten patients (22%) had postoperative complications, and there was one death. CONCLUSION: Bezoar-induced SBO is uncommon and remains a diagnostic and management challenge. It should be suspected in patients with an increased risk of formation of GIB, such as previous gastric surgery, poor dentition, and a suggestive history of increased fibre intake. We advocate that CT imaging be performed early in these at-risk patients and in patients presenting with SBO with or without a history of abdominal surgery in order to reduce unnecessary delays before appropriate surgical intervention. PMID- 17420962 TI - Differential expressions of cyclin D1 associated with better prognosis of cancers of ampulla of Vater. AB - BACKGROUND: Periampullary cancers, the incidence of which increases gradually with industrialization, still pose a significant challenge to clinicians and researchers. Specifically, the role of cell-cycle proteins and tumor suppressor genes in these cancers is not yet clear. Recent studies have revealed that genes and proteins related to cell cycle and apoptosis regulation may be involved in pancreatic carcinogenesis. METHODS: Tissue samples were obtained from patients with periampullary cancers who underwent surgery at the National Taiwan University Hospital without receiving previous chemotherapy or radiation therapy. All periampullary cancer tissue samples were examined by a pathologist, who was unaware of the parameters to be investigated. A total of 68 patients with periampullary cancers (29 ampulla of Vater cancers (AVCs) and 39 pancreatic ductal cancers (PDCs), including various stages and histological subtypes, were enrolled. The relevant demographic and clinicopathological information was obtained from medical records. RESULTS: Cell-cycle proteins, including p16, Rb, cyclin D1, p53, and E2F1, were analyzed by immunohistochemical staining. Here, significant differences were noted between AVCs and PDCs with regard to the expression of cyclin D1. This corresponded to a poor prognosis in PDCs (P < 0.05); AVCs, on the other hand, showed a relatively high survival rate. There is no obvious statistical difference between the 2 groups with regard to the expression of p16, Rb, p53, and E2F1. The study also revealed that cyclin D1 plays different roles in the carcinogenesis of AVCs and PDCs. CONCLUSIONS: The expression of cyclin D1 is more often correlated with prognosis in AVCs than in PDCs, and may serve as a biomarker for the disease. PMID- 17420963 TI - [The feasibility of a registry of dementias: clinical features and diagnostic coverage]. AB - INTRODUCTION AND AIM: Classic epidemiological studies do not allow to know the dementia patterns of derivation and diagnosis in a defined territory. This information is fundamental for the planning and distribution of the sanitary and social resources to a medium-to-long term. The results of a pilot-registry program for dementia cases based on the population surveillance principles is presented. MATERIALS AND METHODS: Consecutive and standardized registry of the incident dementia diagnoses of a memory unit during biennium 2004-2005. RESULTS: 670 new cases of dementia, of which 74% corresponded to Alzheimer's disease, were registered. The presenile dementias were 10.3% of the cases. The mean time between the beginning of the symptoms and the clinical diagnosis was of 2.4 years and the severity of the dementia was mild in 60.0%. 90.1% of the cases lived in their homes or in a relative's home. The hypertension, the diabetes mellitus and the antecedents of depressive disease were the more frequent pathological antecedents (> 20%). The diagnostic coverage based on the estimated dementia cases was 75% for the memory unit's reference territorial area. The diagnostic coverage for the health region of Girona was 38%. CONCLUSIONS: The registered data show the viability and validity of the proposed registry program for dementia cases. Nowadays the registry program is able to extend the diagnostic coverage all over the health region of Girona. PMID- 17420964 TI - [Neonatal polysomnography in newborn infants with severe birth asphyxia]. AB - INTRODUCTION: Neonatal polysomnography studies (electroencephalogram, electrooculography, body movements, cardiorespiratory frequencies) were performed in 101 newborn full-term infants diagnosed with severe birth asphyxia. PATIENTS AND METHODS: To analyse results, the sample was divided into two groups, depending on whether hypoxic-ischaemic encephalopathy (HIE) had occurred or not. The results of the polysomnography studies were correlated with those from the full-term neurological examination and the sequelae from the neurological development during the first two years of life. RESULTS: Significant correlations were obtained among the variables that were studied. The normality observed in the electrophysiological study in the group of patients with severe asphyxia without HIE was associated with a full-term neurological examination and with a neurological development that has progressed in a satisfactory manner. In the group of patients with grade II HIE there was a predominance of severe alterations in the full-term neonatal polysomnography study, which were significantly correlated with the pathological full-term neurological examinations and serious sequelae from neurological development. CONCLUSIONS: It has been proved that neonatal polysomnography studies are a valuable aid in evaluating the neurological status of newborn infants in a critical condition and in predicting the sequelae of neurological development in the first two years of life. Further research should be aimed at determining the effects exerted by the daily administration of medication (sedatives, anaesthetics and antiepileptic agents) in critical newborn infants on electrical activity in the brain and the cyclic structuring of the different phases of sleep. PMID- 17420965 TI - [Ocular myasthenia gravis: diagnostic aspects and evolution]. AB - INTRODUCTION: Myasthenia gravis lacks a diagnostic gold standard, so diagnosis is supported by the findings of several tests. Conversion from ocular myasthenia gravis (OMG) to a more widespread disease is frequently early. AIMS. To describe and compare the clinical data and findings of the diagnostic tests of patients with OMG and those with a different diagnosis, and to know the conversion rate from OMG to generalized myasthenia. PATIENTS AND METHODS: Descriptive and analytic retrospective study of 44 patients referred for neurophysiological testing through 4 years because of suspected OMG. RESULTS: 12 patients (27%) were diagnosed as having OMG. Evolution time prior to diagnosis tended to be shorter in OMG patients. Isolated ptosis or combined with diplopia was more frequent in OMG, while isolated diplopia was so in other diagnoses (p = 0.003). No thymoma was found. SFEMG jitter of facial muscles was abnormal, including blocking, in all OMG patients (8/8) and normal in the rest (30/30). Edrophonium test was positive in all OMG patients (7/7) and doubtful in another one (1/7). Anti-AChR titers were initially positive in 10/11 OMG patients and 0/17 with other diagnoses. Aponeurotic ptosis and strabismus were the most frequent non myasthenic etiologies. OMG showed an early generalization in two patients who developed dysarthria. CONCLUSIONS: The low ratio of diagnostic confirmation suggests that in the face of ptosis or diplopia diagnostic testing with a high sensibility for OMG is favoured. Jitter showed the best initial diagnostic performance. PMID- 17420966 TI - [The use of evoked potentials in the follow-up and prognosis of patients in coma following severe traumatic brain injury]. AB - INTRODUCTION: When dealing with patients in coma, it can be very useful to have early access to objective tests that provide information about their clinical status and can help to establish a prognosis for survival and functional recovery. The value of evoked potentials (EP) in such cases is already well known, although only techniques that assess the neurological status have been used. AIM: To determine the usefulness of different types of evoked potentials as a means of reaching a short and medium term prognosis for survival and functional recovery in patients in coma as a result of traumatic brain injury (TBI). PATIENTS AND METHODS: Our study involved patients with an admission Glasgow coma score of 8 or below due to severe TBI. During the first 24 hours the following tests were carried out on patients: somatosensory EP (SSEP), brainstem auditory EP, visual EP (VEP) obtained using stimulation goggles and mismatch negativity, as well as an imaging test (computerised axial tomography). The tests were repeated at one week and one month, and each patient was evaluated by means of the Glasgow scale and the Karnofsky scale. RESULTS: We studied 22 patients (mean age: 39.95 years; interval: 15-84 years), 20 of them were males. The highest number of deaths occurred during the first week. There was a strong correlation between survival and the results from the SSEP and VEP. CONCLUSIONS: SSEP and VEP performed on admission were a valuable aid for reaching a short and medium term prognosis for survival of patients in coma due to TBI, although they were not effective in evaluating a prognosis for functional recovery. PMID- 17420967 TI - [Unilateral apraxia of eyelid closure in ischemic stroke: role of the right hemisphere in the emotional gesture communication]. AB - INTRODUCTION: Apraxia of eyelid closure (AEC) is an infrequent disorder that is characterised by the inability to close the eyelids on command, although spontaneous blinking and reflex shutting of the eyes is preserved. Very few cases of unilateral AEC have been reported and no long-term follow-ups have been carried out. We report the case of a patient with unilateral AEC that was followed up over a 3-year period and also discuss the role played by the right hemisphere in this disorder. CASE REPORT" We examined the case of a 67-year-old right-handed male who was admitted because of a parietotemporal infarction with extension into the subcortex. A few days after the stroke the patient reported the inability to close his left eye on command, although he was still able to blink spontaneously and the blink and visual threat reflexes were preserved. This deficiency was associated to a dense hemiparesis on the left side and notable aprosodia affecting language. At 3 years' follow-up the AEC had not improved significantly. CONCLUSIONS: There are data to suggest that the right hemisphere is related to emotional perception and expressiveness, as well as the regulation of language prosody. Likewise, symbolic anthropology stresses the importance of winking as a gesture involved in non-verbal communication. These data suggest that AEC may be a consequence of a dysfunction of the role played by the right hemisphere in emotional expressiveness through gestures. The association with language aprosodia could support this hypothesis. PMID- 17420968 TI - [The metabolic regulation of cerebral microcirculation]. AB - AIM: To analyse the mechanisms involved in the microvascular metabolic regulation of cerebral blood flow. The article outlines the neuronal metabolism and mechanisms involved in functional hyperaemia and examines the contractile properties of brain microvasculature. It also discusses the role played by ion channels in pericytes and vascular smooth muscle and describes the signalling pathways involved in arteriolar and capillary vasodilatation or vasoconstriction. DEVELOPMENT: The blood-brain barrier and the close functional relations between neurons and astrocytes give rise to nerve tissue properties such as functional hyperaemia. In this mechanism the astrocytes act as a structural and functional 'bridge' between neurons and brain capillaries, and respond to synaptic activity by releasing vasoactive compounds, above all vasodilators. The metabolites derived from arachidonic acid, such as prostaglandins and epoxyeicosatrienoic acids, as well as the compounds that have traditionally been involved, such as nitric oxide and prostacyclin, are especially important. These substances are capable of extending to the capillaries and arterioles, where they alter the membrane potential and the contractibility of the pericytes and the smooth muscle of vessels. CONCLUSIONS: The functional interaction among neurons, astrocytes and capillaries in the central nervous system (called the 'neurovascular unit') is essential for the regulation of cerebral blood flow, since it links neuron-glial cell metabolic activity to the supply of energetic substrates from the microcirculation. Within this functional unit, astrocytes play a vital role by releasing vasoactive substances that are derived from or produced by neuronal activity. PMID- 17420969 TI - [Rehabilitation therapy in multiple sclerosis]. AB - AIM: To review the rehabilitation therapy and rating scales used in multiple sclerosis (MS), as well as the clinical studies conducted to evaluate the effectiveness of rehabilitation therapy and the most common symptoms. DEVELOPMENT: MS has a number of very specific features that will have to be taken into account when undertaking rehabilitation therapy. It is a disease that progresses in a fluctuating and unpredictable manner and, to date, there is no treatment to cure it. There is a possibility of spontaneous recovery, especially in the early stages of the disease. Evaluation of the effectiveness of rehabilitation in MS must be performed on different levels that take into account both particular aspects of rehabilitation and an overall assessment of the treatment being provided. Most studies report improvements in disability and quality of life, while deficit remains unchanged. Any improvements that are achieved tend to decline over time, which makes it necessary to perform a follow up after rehabilitation therapy ends. Certain symptoms such as fatigue, spasticity, ataxia and sphincteric disorders will benefit from suitable pharmacological treatment applied together with the rehabilitation therapy. CONCLUSIONS: The value of rehabilitation therapy for MS patients lies in its ability to improve disability and the quality of life; since its effects decline over time, regular check-ups will be essential. Proving the effectiveness of rehabilitation in a heterogeneous pathology that develops progressively, such as MS, is a complex matter and further research with better designs and rating scales needs to be conducted. PMID- 17420970 TI - [Neuroimaging and drug addiction: neuroanatomical correlates of cocaine, opiates, cannabis and ecstasy abuse]. AB - INTRODUCTION: Drug abuse is related to neurocognitive alterations linked to the functioning of several areas of the brain. The application of advanced neuroimaging techniques has allowed important advances to be made in research being conducted on why stable disorders are produced in the brain mechanisms responsible for the cognitive processes and on determining exactly what mechanisms drugs of abuse are involved in. AIM. To discuss the evidence available regarding the existence of alterations in the brains of consumers of the most prevalent drugs of abuse in western societies, that is, cocaine, opiates, ecstasy and cannabis. DEVELOPMENT: We review the main neuroimaging studies that have detected alterations in the brain structure and functioning of drug abusers. Likewise, we also discuss the findings from functional neuroimaging studies that have analysed patterns of brain activation associated to specific cognitive operations, such as memory or the executive functions. CONCLUSIONS: Drug abusers present significant alterations in extensive areas of the cortex (especially in the frontal and temporal cortex), subcortex (amygdala, hippocampus and insular cortex) and basal regions (striatum). These alterations are associated with abnormal patterns of activation during cognitive memory tasks, inhibition and decision-making. Cocaine abusers present more pronounced and generalised alterations in the brain. By combining neuropsychological and neuroimaging findings is has been shown that the motivational, memory and executive control processes can play a key role in rehabilitating drug addicts. PMID- 17420971 TI - [Meningoencephalocele as the causation of recurring meningitis in infants]. PMID- 17420972 TI - [Motor neuron disease secondary to lead poisoning]. PMID- 17420973 TI - [Vertical nystagmus secondary to the epidural administration of morphine]. PMID- 17420974 TI - [Acute cerebrovascular diseases: a challenge for today's health care systems]. PMID- 17420975 TI - [Strokes in the district hospital: please call the neurologist]. PMID- 17420977 TI - Predictors of comprehensive surgical treatment in patients with ovarian cancer. AB - BACKGROUND: Providing appropriate surgical treatment for women with ovarian cancer is one of the most effective ways to improve ovarian cancer outcomes. In this study, the authors identified factors that were associated with a measure of comprehensive surgery, so that interventions may be targeted appropriately to improve surgical care. METHODS: Using Healthcare Cost and Utilization Project hospital discharge data from 1999 to 2002 for 9 states, the authors identified 10,432 admissions of women who had an International Classification of Disease, 9th Revision (ICD-9) primary diagnosis of ovarian cancer and who had undergone oophorectomy. Based on National Institutes of Health Consensus Panel recommendations, surgeries were categorized as comprehensive by using ICD-9 diagnosis and procedure codes. Logistic regression analysis using data from 5 states with a full set of variables (n = 6854 patients)was used to identify factors that were associated with the receipt of comprehensive surgical care. RESULTS: Overall, 66.9% of admissions (range, 46.3-80.8% of admissions) received comprehensive surgery. Factors that were associated independently with comprehensive surgical care included age (ages 21-50 years vs ages 71-80 years or > or = 81 years), race (Caucasian vs African American or Hispanic), payer (private insurance vs Medicaid), cancer stage (advanced vs early), annual surgeon volume (low/medium [2-9 surgeries per year] or high [>10 surgeries per year] vs very low [1 surgery per year]), and surgeon specialty (gynecologic oncologists vs obstetrician gynecologists or general surgeons). Among nonteaching hospitals, medium-volume hospitals (10-19 ovarian cancer surgeries per year) and high-volume hospitals (> or = 20 surgeries per year) had significantly higher comprehensive surgery rates than low-volume facilities (1-9 surgeries per year). Volume did not influence comprehensive surgery rates in teaching hospitals. CONCLUSIONS: Many women with ovarian cancer, especially those in poor, elderly, or minority groups, are not receiving recommended comprehensive surgery. Efforts should be made to ensure that all women with ovarian cancer, especially those in vulnerable populations, have the opportunity to receive care from centers or surgeons with higher comprehensive surgery rates. PMID- 17420978 TI - PSI3: an open-source Ab Initio electronic structure package. AB - PSI3 is a program system and development platform for ab initio molecular electronic structure computations. The package includes mature programming interfaces for parsing user input, accessing commonly used data such as basis-set information or molecular orbital coefficients, and retrieving and storing binary data (with no software limitations on file sizes or file-system-sizes), especially multi-index quantities such as electron repulsion integrals. This platform is useful for the rapid implementation of both standard quantum chemical methods, as well as the development of new models. Features that have already been implemented include Hartree-Fock, multiconfigurational self-consistent field, second-order Moller-Plesset perturbation theory, coupled cluster, and configuration interaction wave functions. Distinctive capabilities include the ability to employ Gaussian basis functions with arbitrary angular momentum levels; linear R12 second-order perturbation theory; coupled cluster frequency dependent response properties, including dipole polarizabilities and optical rotation; and diagonal Born-Oppenheimer corrections with correlated wave functions. This article describes the programming infrastructure and main features of the package. PSI3 is available free of charge through the open source, GNU General Public License. PMID- 17420979 TI - Role of radiotherapy in ductal (endometrioid) carcinoma of the prostate. AB - BACKGROUND: Ductal carcinoma of the prostate is a rare variant of prostate cancer that presents most commonly with obstructive urinary symptoms or hematuria. This case series of 6 patients is the first to report the outcome of ductal carcinoma treated with external beam radiotherapy. METHODS: A retrospective review was performed of patients treated between 1980 and 2006 at Fox Chase Cancer Center, Philadelphia, Penn. Six patients were identified with ductal carcinoma. RESULTS: Five of the 6 patients were treated definitively and the sixth patient was treated at recurrence 3 years after a radical prostatectomy. Patient ages ranged from 66-80 years and the initial prostate-specific antigen (iPSA) ranged from 1.69-100.3 ng/mL. Three patients had a mixed acinar and ductal carcinoma, 2 with a Gleason score (GS) of 8 and 1 with a GS of 7. Of the patients treated definitively, 4 had clinical stage T2A-T2C and 1 had clinical stage T1B. Definitive radiotherapy was delivered to the prostate with doses between 72 Gy and 78 Gy. Pelvic lymph nodes were treated in all patients. One patient was treated postradical prostatectomy to the prostate bed to a dose of 60 Gy. Adjuvant androgen deprivation was given in 5 of the patients. Two of the patients died from metastatic disease at 1.4 and 7.1 years after treatment. The remaining 4 patients remain alive between 3.2 and 4.8 years from treatment, with 3 patients biochemically without evidence of disease. No patients have developed a local recurrence. CONCLUSIONS: Ductal carcinoma of the prostate may be treated effectively with external beam radiotherapy. Aggressive management is indicated, even with low-volume metastatic disease. PMID- 17420980 TI - Renal cell carcinoma clinically involving adjacent organs: experience with aggressive surgical management. AB - BACKGROUND: Historically, patients with nonmetastatic renal cell carcinoma (RCC) involving adjacent organs have been considered inoperable and incurable. The oncologic efficacy of an aggressive surgical approach was evaluated in a selected subpopulation of RCC patients. Further, an attempt was made to define the clinical and pathologic characteristics predictive of surgical failure. METHODS: With Institutional Review Board approval, the institutional nephrectomy database of 3470 patients treated at MD Anderson Cancer Center from 1990 to 2006 was searched for RCC patients treated with radical nephrectomy and resection of at least 1 adjacent organ thought to be directly involved by RCC. Patients with nonmetastatic RCC and a minimum follow-up of 6 months were included in the analysis. RESULTS: In all, 30 patients with clinical T4NxM0 RCC and median follow up of 32.3 months (range, 8.5-140.1) met the study inclusion criteria and comprise the dataset for the analysis. On pathologic evaluation 60% of patients were clinically overstaged, as only 12 (40%) of 30 patients demonstrated direct invasion into adjacent organs resected. None of the clinical tumor characteristics predicted a finding of pathologic T4 RCC. Nodal involvement and pathologic T stage were significant independent predictors of disease recurrence (hazard ratio [HR] 3.726, P = .043, and HR 2.414, P = .045, respectively) and cancer-specific survival (HR 17.145, P = .002, and HR 3.791, P = .024, respectively). Disease recurred in 11 of 18 (61.1%) of 0.05). It appears that the risk of reduced lung function was greater the younger the patient in therapy. CONCLUSION: Chemotherapy or chemo-radiotherapy-induced pulmonary sequalae in childhood may remain asymptomatic for many years. PMID- 17420992 TI - Initial testing (stage 1) of the proteasome inhibitor bortezomib by the pediatric preclinical testing program. AB - BACKGROUND: Bortezomib is a proteasome inhibitor that has been approved by FDA for the treatment of multiple myeloma and that has completed phase 1 testing in children. The purpose of the current study was to evaluate the antitumor activity of bortezomib against the in vitro and in vivo childhood cancer preclinical models of the Pediatric Preclinical Testing Program (PPTP). PROCEDURES: Bortezomib was tested against the PPTP in vitro panel at concentrations ranging from 0.1 nM to 1.0 microM and was tested in vivo at a dose of 1 mg/kg for a planned duration of 6 weeks. RESULTS: Bortezomib was uniformly active against the PPTP's in vitro panel, with a median IC(50) of 23 nM and with a steep dose response curve. The four acute lymphoblastic leukemia (ALL) cell lines had significantly lower IC(50) values compared to the remaining lines of the in vitro panel. Limited in vivo activity was observed for bortezomib against the solid tumor xenografts tested, with one line meeting criteria for intermediate activity for the time to event measure and with the remaining lines showing low activity for this measure. Bortezomib demonstrated in vivo activity against the ALL panel, inducing two complete and two partial responses among seven evaluable lines. CONCLUSIONS: Administered at its MTD in mice, bortezomib demonstrated activity against selected lines of the PPTP's ALL in vivo panel. Further studies are indicated to determine the activity of bortezomib when combined with standard agents to treat childhood ALL. PMID- 17420994 TI - Closure of guide wire-induced coronary artery perforation with a two-component fibrin glue. AB - Perforation or rupture of a coronary artery with subsequent pericardial effusion and cardiac tamponade is a potentially life-threatening complication of percutaneous coronary intervention (PCI). Several emergency treatment strategies exist to close the perforation including reversal of anticoagulation, prolonged balloon inflation, implantation of stent grafts, local injection of thrombogenic molecules, placement of microcoils, or open heart surgery. Here we report on a 66 year-old patient who underwent urgent PCI for acute stent thrombosis in the proximal LAD. The artery was reopened, a new stent implanted successfully, and a GPIIb/IIIa-antagonist was given. Shortly thereafter the patient suffered from cardiac tamponade requiring pericardiocentesis and pericardial drainage. The coronary angiogram indicated a severe guide wire-induced perforation and pericardial effusion originating from a distal diagonal branch segment. Prolonged balloon inflation did not stop the leakage. Therefore the monorail balloon was exchanged for an over-the-wire balloon. A two-component commercial fibrin glue consisting of fibrinogen and thrombin was rapidly but separately injected through the wire channel of the balloon into the distal segment of the diagonal branch. The coronary leak was successfully closed and the patient recovered quickly. In comparison with the previously reported cases of thrombin injection important differences should be noticed: (1) a two-component hemostatic seal was used without reversal of anticoagulation, (2) rapid injection instead of prolonged infusion of the hemostatic drugs was performed, and (3) the rescue technique was applied in a cath lab that routinely uses monorail catheter systems. Therefore we consider this a novel and effective approach for closure of coronary ruptures. PMID- 17420995 TI - Coronary artery fistulas: how to manage them. AB - Coronary artery fistulas are rare persisting vascular connections from a coronary artery to a cardiac chamber or major central blood vessel. The true incidence is difficult to discern because at least 75% may be asymptomatic and clinically undetectable until an echocardiogram or coronary arteriogram is performed. Small coronary artery fistulas that are not clinically detectable are not clearly associated with significant long term complications. Medium or large fistulas are associated with significant long term problems including angina, arrhythmias, myocardial infarction, endocarditis, and progressive dilation. Treatment options include surgical and catheter approaches to significant fistulas. The long term outlook after fistula closure is not well defined and deserves further study. PMID- 17420996 TI - Endurance running and digit ratio (2D:4D): implications for fetal testosterone effects on running speed and vascular health. AB - There is anatomical and physiological evidence that endurance running (ER), i.e., running one or more kilometers using aerobic metabolism, originated early in the evolution of Homo, and the consequences of early selection for ER may be important in modern Homo. Here we examine ER performance in competitive ER. ER is sex dependent such that men tend to run faster than women, and the influence of sex on ER suggests that it may be modified by testosterone (T). It is shown that a putative proxy for prenatal T, the ratio of the length of the 2nd and 4th digits (2D:4D), is correlated with ER. Thus performance in training for ER was associated with high prenatal T, as measured by low 2D:4D, in both men and women. In cross-country races from 1 to 4 miles, 2D:4D explained about 25% of the variance in both male and female ER. Therefore, speed in ER was dependent on a proxy for prenatal T. 2D:4D correlates with performance in sport and exercises, which test a mix of strength and fitness, but the associations are in general quite weak with 2D:4D accounting for less than 10% of the variance in performance. Our finding that 2D:4D explains about 25% of the variance in ER suggests that prenatal T is important in determining efficiency in aerobic exercise. Early populations of Homo may have been strongly selected for ER and high prenatal T. The implications of this for patterns of predisposition to cardiovascular disease in modern Homo are discussed. PMID- 17420997 TI - Microlevel determinants of blood pressure among women of two ethnic groups in a periurban area of Kolkata city, India. AB - Blood pressure (BP) trends vary cross-culturally, and studies on the risk factors associated with hypertension are limited in periurban regions of India. This study was conducted to examine the effect of socioeconomic factors (income, expenditure, activity time) and anthropometric measurements (skinfolds of biceps, triceps, subscapular, supra iliac, and body mass index) on 102 Munda (tribe) and 135 Pod (caste) women of childbearing age in a periurban area of Kolkata city. Results indicate that Munda women had significantly higher diastolic BP in the 30+ age group. However, no difference in the systolic and diastolic BP was observed between the two groups, when the socioanthropometric factors were controlled as covariates. Also, according to the JNC (JNC, 2004, The JNC 7 Report NIH Publication No. 04-5230) classification of hypertension, no significant difference in hypertension was noted between the women of the two social groups. But the risk factors associated with hypertension varied between the two ethnic groups. Expenditure on alcohol and activity time was associated with hypertension among the Munda, while body mass index was significantly associated with hypertension among the Pod women. Alcohol consumption is a rare phenomenon among Indian women. Yet, Munda women in this transitional periurban environment, in spite of high poverty were more inclined to spent their earnings in alcohol consumption (due to their cultural preferences), increasing the risk of hypertension in their childbearing age. PMID- 17420998 TI - Y-chromosome lineages in Sao Tome e Principe islands: evidence of European influence. AB - The Y-chromosome haplogroup composition of the population of Sao Tome e Principe (STP) archipelago was analyzed using 25 biallelic markers and compared with populations of different origins from Europe, Africa, and the Middle East. Two main Y-chromosome haplogroups were found: E3a, very common among sub-Saharans accounts for 84.2% of the paternal lineages and R1b, typical of West Eurasia, represents 8.7% of the overall male population. Nevertheless, we detected in the population of STP a significant heterogeneous distribution of R1b among the two main ethnic groups of the archipelago: Forros (10.3%) and Angolares (6.6%). Together, haplogroups known to be prevalent in West Eurasia reach 12.5% of the chromosomes analyzed unequally distributed among the two groups: Forros present 17.7% while Angolares display only 8.2% of west Eurasian haplogroups. Our findings suggest that, despite its sub-Saharan genetic background, a relevant contribution of European paternal lineages is present in nowadays STP population. This influence has shown to be stronger in Forros than in Angolares, which could be explained by the social isolation that these have last experienced through their history. PMID- 17420999 TI - Pre-existing left pulmonary artery stenosis and other anomalies associated with device occlusion of patent ductus arteriosus. AB - A retrospective analysis was performed on 200 consecutive patients who underwent cardiac catheterization for occlusion of Patent Ductus Arteriosus (PDA) at a single center by a single operator. Four significant anomalies were observed: pre existing Left Pulmonary Artery (LPA) stenosis, left recurrent laryngeal nerve (LRLN) injury, electrocardiogram (EKG) changes, and aorto-pulmonary (AP) collateral arteries. The observation of pre-existing LPA stenosis, marked EKG changes, and permanent LRLN injury have not been previously reported. Incidence, etiology, and clinical significance of these anomalies are discussed with specific new recommendations for the prevention of LRLN injury and occlusion of AP collaterals. PMID- 17421000 TI - Enzyme immunoassay for total immunoglobulin E in dried blood spots. AB - Elevated circulating levels of total immunoglobulin E (IgE) are associated with both allergic disease and repeated macro-parasitic infections. Population-based research on IgE has been limited by the logistical constraints associated with obtaining and processing venipuncture blood samples. In this short report, we present an enzyme immunoassay protocol for quantifying circulating total IgE levels in capillary whole blood, collected from a finger prick and dried on filter paper. The assay demonstrated acceptable levels of accuracy, precision, and reliability. IgE remained stable at room temperature for only 2-4 days and degraded rapidly at higher temperatures suggesting that samples should be refrigerated or frozen within 1-2 days of collection. It is hoped that the relative ease of blood spot collection will expand opportunities for population based research on IgE. PMID- 17421001 TI - New saliva DNA collection method compared to buccal cell collection techniques for epidemiological studies. AB - Epidemiological studies may require noninvasive methods for off-site DNA collection. We compared the DNA yield and quality obtained using a whole-saliva collection device (Oragene DNA collection kit) to those from three established noninvasive methods (cytobrush, foam swab, and oral rinse). Each method was tested on 17 adult volunteers from our center, using a random crossover collection design and analyzed using repeated-measures statistics. DNA yield and quality were assessed via gel electrophoresis, spectophotometry, and polymerase chain reaction (PCR) amplification rate. The whole-saliva method provided a significantly greater DNA yield (mean +/- SD = 154.9 +/- 103.05 microg, median = 181.88) than the other methods (oral rinse = 54.74 +/- 41.72 microg, 36.56; swab = 11.44 +/- 7.39 microg, 10.72; cytobrush = 12.66 +/- 6.19, 13.22 microg) (all pairwise P < 0.05). Oral-rinse and whole-saliva samples provided the best DNA quality, whereas cytobrush and swab samples provided poorer quality DNA, as shown by lower OD(260)/OD(280) and OD(260)/OD(230) ratios. We conclude that both a 10 ml oral-rinse sample and 2-ml whole-saliva sample provide sufficient DNA quantity and better quality DNA for genetic epidemiological studies than do the commonly used buccal swab and brush techniques. PMID- 17421002 TI - Genetics of anthropometric asymmetry in an Indian endogamous population-Vaidyas. AB - To understand the genetics of Fluctuating Asymmetry (FA) and Directional Asymmetry (DA), the present study comprised 14 bilateral morphometric traits from 200 Vaidya families including 824 individuals (of two generations) from North 24 Parganas, West Bengal. The statistical analysis included: Regression analysis to remove the age effect, Familial correlation, Heritability estimation, Principal Component Analysis and Segregation Analysis (SA) using genetic model test. The obtained results revealed little effect of genetic factor and considerable amount of environmental influence on anthropometric asymmetry. The results support the idea postulated by several previous authors that FA provides a measure of developmental instability in man. The contribution of heredity on these asymmetric variables is not unimportant but that of the common environment is very substantial. The magnitude of heritability of DA traits is slightly higher than that of FA traits. Five principal factors were detected from these asymmetric traits (three factors are on asymmetry on length, head, and breadth; while last two factors represent the asymmetry of diameters). SA did not suggest any evidence of major gene contribution. But the involvement of minor genes or polygenes could not be discarded. As the study on SA of asymmetry in man is limited, similar other studies are needed to confirm the result of the present study. PMID- 17421003 TI - Validation of hand-held bioelectrical impedance analysis with magnetic resonance imaging for the assessment of body composition in overweight women. AB - Methods of assessing body composition suitable for use in clinical trials should be accurate, reliable, and easy to perform. One such technique routinely implemented is hand-held bioelectrical impedance analysis (BIA). The validity of this method, however, in body composition assessment of overweight women is not known. The aim of this study was to validate the hand-held BIA technique with magnetic resonance imaging (MRI) for the assessment of body composition in overweight women. Fat mass, percent fat mass, fat-free mass, and percent fat-free mass values estimated by hand-held BIA were compared to those measured by MRI. Thirty-one Caucasian women (50.1 +/- 8.2 years, body mass index of 26.9 +/- 3.1 kg/m(2)) participated in the study. BIA measurements were highly reproducible (technical error (TE) was 0.06 +/- 0.07 kg for fat mass and 0.08 +/- 0.11% for percent fat mass), but were significantly different (P < 0.0001) for each body composition parameter when compared to MRI. BIA underestimated fat mass by 2.3 +/ 3.3 kg and percent fat mass by 5.6 +/- 3.9%. Likewise, BIA overestimated fat free mass by 7.4 +/- 2.7 kg and percent fat free mass by 5.6 +/- 3.2%. No relationship between the bias and the mean of the two measurements was noted suggesting that bias is not related to measurement size. Although hand-held BIA gives reproducible findings, the bias noted for all body composition parameters puts into question the validity of this regional impedance device for use in clinical trials in overweight women. PMID- 17421004 TI - Index-to-ring finger length ratio (2D:4D) predicts levels of salivary estradiol, but not progesterone, over the menstrual cycle. AB - We tested the association between the index-to-ring finger length ratio (2D:4D) and ovarian steroid hormone concentrations measured over the course of a menstrual cycle in the saliva of 38 young women. Estradiol levels were positively associated with right-hand, but not left-hand, 2D:4D, and also with the difference between right- and left-hand 2D:4D. None of these measures predicted progesterone level. PMID- 17421005 TI - Grip strength and digit ratios are not correlated in women. AB - Fink et al. (2006) have reported that men with higher grip strength (GS) have lower digit ratios (2D:4D), interpreting this as evidence for organizational effects of prenatal androgens on strength. In this study, I attempted to replicate their finding with 99 women. I found no evidence that digit ratios were associated with GS, suggesting that 2D:4D is not associated with GS in women. The null findings are discussed in light of gender and statistical considerations. PMID- 17421006 TI - Prepregnancy weight, inappropriate gestational weight gain, and smoking: Relationships to birth weight. AB - This study was designed to test predictors of infant birth weight based on categories of prepregnancy body mass index (BMI), gestational weight gain, and smoking. Data were collected retrospectively from records of 233 mother-infant pairs enrolled in the Siouxland Women, Infants, and Children (WIC) Program in Sioux City, Iowa. Prepregnancy BMI and gestational weight gain were coded according to Institute of Medicine guidelines. Smoking behavior was coded based on reported smoking during the last 3 months of pregnancy. Multiple regression analysis was used to test predictors of infant birth weight. Forty-two percent of women gained more weight than recommended, and 16% gained less than recommended. Based on prepregnancy BMI, women were classified as underweight (12%), overweight (16%), or obese (31%). Twenty-four percent of mothers reported smoking late in gestation. Higher birth weight was predicted by prepregnancy obesity (+144 g). Lower infant birth weight was predicted by lower (-162 g) and higher (-153 g) than recommended weight gain, and by lower (-299 g) and higher (-168 g) levels of smoking. Depression of birth weight among women who gained excess weight may relate to inadequate early weight gain or pregnancy complications. More research is needed regarding physiological consequences of these maternal factors and their associated demographic risk factors. PMID- 17421007 TI - HLA genes and surnames show a similar genetic structure in Lombardy: does this reflect part of the history of the region? AB - Lombardy, in northern Italy, is the most populated and industrialized Italian region. We attempt to study its genetic structure with two independent sets of data: HLA allele frequencies and surnames. According to our results, it is plausible to deduce that ancient history, more than genetic isolation and drift, may have contributed to the present genetic structure of Lombardy. The hypothesis seems to be confirmed by the results of the cluster analysis of the 11 provinces of the region, which was performed using two different types of markers. Both genes and surnames show approximately the same structure. Not only Celts but also ancient Ligurians (and Etruscans) probably shaped the region into the present three clusters in which the 11 provinces appear to be genetically structured. In particular, an ancient historic, archaeological, and linguistic boundary, along the Adda River, seems to be preserved in present-day Lombardy's population structure. PMID- 17421008 TI - Size at birth and growth trajectories to young adulthood. AB - Few studies in developing countries follow growth trajectories from birth to adulthood. Such studies are important because size at birth and postnatal growth affect risk of chronic disease in adulthood. This study examines the inter relationships of maternal factors during pregnancy, infant birth weight and length, early postnatal growth, and young adult height, weight, BMI, and skinfold thicknesses, with particular attention to patterns of growth associated with increased chronic disease risk. Women were recruited in pregnancy, and offspring were followed from birth to age 21 in the community-based Cebu (Philippines) Longitudinal Health and Nutrition Survey. Birth weight and length are independently, positively associated with height, BMI and sum of skinfolds in young adult males and females, and inversely associated with the subscapular to triceps ratio in males only. The effects of size at birth on adult size were modified by birth order, and remained significant after adjusting for maternal nutritional status, socioeconomic status at birth and throughout the growth period, and maturation. Early postnatal growth was strongly influenced by BMI at birth, with rapid early infant weight gain associated with thinness. The growth pattern of the at-risk group most often associated with increased risk of chronic disease (small at birth, relatively heavy as an adult), was characterized by more rapid growth in the first 4 postnatal months. The high level of inter-relatedness of maternal nutrition in pregnancy, prenatal growth, and postnatal growth emphasizes the need to consider the full growth trajectory in studies of developmental origins of adult disease. PMID- 17421009 TI - Single nucleotide polymorphisms in alcohol dehydrogenase genes among some Indian populations. AB - Seven ADH genes, identified until now, located in the long arm of human chromosome 4, produce seven different isozymes involved in the metabolism of ethanol to acetaldehyde. Of the more than 500 SNPs reported in the coding and non coding regions of these genes in the world databases, 11 are more extensively studied. Three SNPs, ADH1B Arg47His (Exon3), ADH1B Arg369Cys (Exon9) and ADH1C Val349Ile (Exon8), are functionally validated in terms of phenotype-genotype correlations and are in specific linkage disequilibrium (LD) with non-coding SNPs. However, the frequency of each SNP and configuration of LD varies among populations. The Indian populations studied were conspicuous by the complete absence of African specific allele ADH1B*369Cys, the negligible frequency of East Asian specific ADH1B*47His allele and the presence of a novel SNP ADH1B A3529G (Intron3). The ADH1C*349Ile was the only functional allele polymorphic with a strong LD block in all the populations studied and the high F(st) value observed for the non-coding ADH1B Rsa1 variant was in conformity with world populations. PMID- 17421010 TI - Secular trends in anthropometric characteristics, physical fitness, physical activity, and biological maturation in Flemish adolescents between 1969 and 2005. AB - In this study, secular trends in anthropometric characteristics, physical fitness, physical activity, and biological maturity over the past 25-35 years in Flemish adolescents were investigated. Representative cross-sectional samples of 12-18-year-old secondary school children (11,899 assessments in boys in 1969 1974, 4,899 girls in 1979-1980, 1,429 boys and 1,772 girls in 2005) and parent offspring pairs tested at approximately the same age during adolescence (55 father-son pairs, mean age fathers = 15.47 years, mean age sons = 15.38 years; 62 mother-daughter pairs, mean age mothers = 16.63 years, mean age daughters = 15.01 years) were used. The cross-sectional data were analyzed in 6 yearly age categories using Wilcoxon rank sum tests. For the parent-offspring data paired t tests, simple linear regressions to adjust for parent-offspring differences in chronological age and multiple linear regressions to adjust for parent-offspring differences in chronological and skeletal age were conducted. The cross-sectional study generally revealed an increase in weight, stature, BMI, skinfolds and trunk extremity index, and a decrease in the performance on several physical fitness tests. In the parent-offspring study, only sons were maturationally advanced compared to fathers. Even after adjustment for parent-offspring differences in chronological age and in chronological and skeletal age, results for stature, trunk-extremity index and physical fitness were generally similar to the cross sectional study. No secular trend was observed for sports participation. The fact that the positive secular trends in weight, BMI, and skinfolds of the cross sectional study were not entirely confirmed in the parent-offspring study is probably due to higher similarity in genetic and familial background, higher socio-economic status, and more health-consciousness of the latter. PMID- 17421011 TI - Focused life history data and linear enamel hypoplasia to help explain intergenerational variation in relative leg length within Taiwanese families. AB - This study first examines the hypothesis that significant intergeneration increases in stature within 85 Taiwanese families in two study locations were primarily the result of increases in subischial leg length. It then evaluates a second hypothesis that independent assessments of the extent of intergenerational change in childhood environments within these families helped account for parent offspring differences in relative leg length. Childhood environments were assessed using two criteria: developmental environment scores derived from life history data and evidence of linear enamel hypoplasia (LEH). Relative leg length was represented in two ways, as either the difference between internally estimated sex-specific z-scores of sitting height and subischial leg length (z(D) = z(SH) - z(LL)) or as relative sitting height (RSH; sitting height/height x 100). Paired Student t tests indicated intergenerational increases in relative leg length were significantly greater than zero in both study locations (P < or = 0.012). The second hypothesis, tested using a hierarchical model with maximum likelihood estimation that allowed for nesting of more than one offspring per family, received support as midparent-offspring differences in composite scores were significantly positively associated with midparent-offspring differences in relative leg length. This was true alone (P = 0.018), and when significant associations with LEH prevalence among mothers and offspring were statistically controlled for (P = 0.010). Evidence also indicated that while the large majority of offspring were taller than their midparental average height (84.3%; 91/108), offspring who were taller were also more likely to have relatively longer legs than by chance alone (Fishers exact, P = 0.027). PMID- 17421012 TI - Mortality experience of Tsimane Amerindians of Bolivia: regional variation and temporal trends. AB - This paper examines regional and temporal trends in mortality patterns among the Tsimane, a population of small-scale forager-horticulturalists in lowland Bolivia. We compare age-specific mortality in remote forest and riverine regions with that in more acculturated villages and examine mortality changes among all age groups over the past 50 years. Discrete-time logistic regression is used to examine impacts of region, period, sex, and age on mortality hazard. Villages in the remote forest and riverine regions show 2-4 times higher mortality rates from infancy until middle adulthood than in the acculturated region. While there was little change in mortality for most of the life course over the period 1950-1989, overall life expectancy at birth improved by 10 years from 45 to 53 after 1990. In both periods, over half of all deaths were due to infectious disease, especially respiratory and gastrointestinal infections. Accidents and violence accounted for a quarter of all deaths. Unlike typical patterns described by epidemiologic transition theory, we find a much larger period reduction of death rates during middle and late adulthood than during infancy or childhood. In the remote villages, infant death rates changed little, whereas death rates among older adults decreased sharply. We hypothesize that this pattern is due to a combination of differential access to medical interventions, a continued lack of public health infrastructure and Tsimane cultural beliefs concerning sickness and dying. PMID- 17421013 TI - Coronary stent assessability by 64 slice multi-detector computed tomography. AB - BACKGROUND: We evaluated the assessability of contemporary stent platforms by 64 slice multi-detector computed tomography (MDCT). METHODS: Patients undergoing coronary stenting were included in a prospective protocol of MDCT imaging within 48 hr of stent implantation. MDCT data were acquired using a "Sensation 64" MDCT scanner (Siemens Medical Solutions, Forchheim, Germany). Stent assessability was assessed by two independent blinded observers and disagreement was resolved by a third observer. Assessability was defined at visualization of the in-stent lumen without influence of partial volume effects, beam hardening, motion, calcification, or contrast to noise limitations. RESULTS: Fifty four stents (Cypher n = 25, Vision/Minivision n = 19, Taxus Express n = 8, Liberte n = 1, Driver n = 1) in 44 patients were included in the study. The two independent observers classified 30 of 54 stents (56%) as assessable. Interobserver reproducibility was good with kappa = 0.66. Stent size was the most important determinant of assessability. Consistently assessable stents were 3.0 mm or larger (85%), whereas those under 3 mm were mostly nonassessable (26%). CONCLUSIONS: Contemporary stent designs evaluated on a 64-slice MDCT scanner showed artifact free assessability only in larger stents. Increase in spatial resolution of MDCT scanners or modifications in stent design will be necessary to noninvasive evaluate stents <3 mm in diameter, where in-stent restenosis is more frequent. PMID- 17421014 TI - Percutaneous coronary intervention in two occluded arms of a saphenous 'Y'-graft: the importance of protecting both of your arms. AB - Percutaneous intervention in saphenous vein grafts (SVG) carries a higher risk of distal embolization than intervention in a native vessel, and use of a distal protection device has been shown to improve the outcomes in SVG interventions. We describe an intervention done in an unexpected 'Y' SVG which required dual distal protection with Filterwires placed in both limbs of the diseased graft and which was performed via a 6 Fr guide catheter. PMID- 17421015 TI - Percutaneous closure after inadvertent subclavian artery cannulation. AB - Accidental insertion of an arterial sheath is an uncommon but potentially serious complication of jugular venous catheterization. When the subclavian artery is inadvertently cannulated, sheath removal can be complicated by significant hemorrhage due to its incompressible location. We report a case of inadvertent insertion of an 8 French sheath into the subclavian artery, which was successfully removed and the puncture site sealed with a collagen-based vascular closure device (Angio-Seal STS Plus). This averted an otherwise emergent open surgical procedure to remove the sheath and repair the subclavian artery in a high-risk patient. PMID- 17421016 TI - Coronary stenting for coronary vasospasm refractory to medical therapy. AB - We describe the case of a 54-year-old male with recurrent chest pain, ST segment elevation, and bradycardia. Coronary vasospasm was confirmed by acetylcholine challenge. After failing medical therapy, stenting of an extensive segment of the right coronary artery has been clinically successful after 24-months follow-up. PMID- 17421017 TI - Three-dimensional modeling of double-stent techniques at the left main coronary artery bifurcation using micro-focus X-ray computed tomography. AB - BACKGROUND: Various double-stent techniques using drug-eluting stents have been proposed to treat the left main coronary artery (LMCA) bifurcation. However, use of these techniques is frequently associated with focal restenosis at the ostium of the left circumflex coronary artery (LCX). OBJECTIVES: To examine the results of double-stent techniques, using a silicon model of the LMCA bifurcation and three-dimensional (3D) reconstruction images created with micro-focus X-ray computed tomography (MFCT). METHODS: Crush, kissing, and modified T stentings were performed with bare metal stents in a LMCA bifurcation model. The stents were then inspected using MFCT at a minimal resolution of 0.06 mm. RESULTS: Gaps in stent apposition to the vessel were observed at the site of stent overlap in the distal LMCA with all stenting techniques. In crush stenting, when the left anterior descending artery stent overlapped the LCX stent, the latter was crushed on the myocardial side of the vessel, and a gap was observed on the nonmyocardial side, at the LCX ostium. When the overlap was reversed, the LCX stent was crushed on the nonmyocardial side and a gap was observed on the myocardial side. In the case of kissing stents, stent overlap created a gap beneath the overlapped portion of the stents. In modified T-stenting, correct positioning of the LCX stent was difficult and MFCT imaging revealed a nonmyocardial gap. CONCLUSIONS: Close apposition of the stent to the vessel at the ostium of the LCX is difficult to achieve at the LMCA bifurcation, regardless of which double-stent technique is employed, due to the site's wide bifurcation angle and complex 3D structure. The distribution of plaque and the bifurcation angle should be considered before double-stent deployment, to avoid leaving a gap over significant plaques. PMID- 17421018 TI - Feasibility and clinical impact of transcatheter closure of interatrial communications after a fenestrated Fontan procedure: medium-term outcomes. AB - OBJECTIVE: This study was to review an institutional experience with transcatheter closure of Fontan fenestrations and its impact on clinical care. BACKGROUND: An interatrial fenestration improves postoperative outcomes in high risk children undergoing a Fontan repair. While technical feasibility has been well defined, the clinical impact of subsequent closure is not well defined. METHODS: Transcatheter closure of a surgically created or additional interatrial communication was attempted in 152 children at a median interval of 13.8 months after surgery. The clinical records were reviewed for demographic and anatomical characteristics, previous surgeries; catheterization data, and status at latest follow-up. RESULTS: Mean oxygen saturation and right atrial pressure increased acutely from 87% +/- 5% to 96% +/- 3% (P < 0.001) and 12 +/- 2 mm Hg to 13 +/- 3 mm Hg (P < 0.001), respectively. Higher systemic venous atrial pressures after occlusion correlated with higher pulmonary artery pressures (P = 0.05) before the Fontan procedure and with higher right (P < 0.001) and left atrial (P = 0.001) and ventricular end-diastolic pressures (P < 0.001) immediately before occlusion. Complications included device malposition in 2 children, 1 child each had an air embolism and post-procedural bleeding, and each self-limiting and 1 child had acute ST elevation in inferior ECG leads because of occlusion of the acute marginal branch which was treated with angioplasty and placement of a stent. At follow-up (median 4.5 years), the mean oxygen saturation was 95% +/- 3%. Residual interatrial leaks were noted echocardiographically in 9%. Two children developed protein-losing enteropathy after fenestration closure. No deaths or strokes were observed in follow-up. CONCLUSIONS: Transcatheter occlusion of Fontan fenestrations is safe with acute and persistent improvements in oxygen saturations. PMID- 17421019 TI - Percutaneous rheolytic thrombectomy for large pulmonary embolism: a promising treatment option. AB - BACKGROUND: Pulmonary embolism (PE) is a common cardiovascular disease with significant mortality. Some patients with large PE are not eligible for current treatment options such as thrombolysis or surgical embolectomy. We report our experience of percutaneous rheolytic thrombectomy (PRT) using the AngioJet system combined with adjunctive local thrombolytic therapy and inferior vena cava (IVC) filter placement to treat massive or submassive PE in patients ineligible for current treatment options. METHODS AND RESULTS: Of the 14 consecutive patients ineligible for thrombolysis or embolectomy treated with PRT, 10 patients had massive PE (6 patients were hypotensive and 4 patients had intractable hypoxemia) and 4 patients had submassive PE. Adjunctive local thrombolysis was performed in 5 patients. An IVC filter was placed in 11 patients. Angiographic success based on Miller score was achieved in 13 patients (92.9%). Procedure success was obtained in 12 patients (85.7%). Procedural mortality occurred in one patient who presented in cardiogenic shock (7.1%) and non-fatal hemoptysis occurred in 1 patient (7.1%). Total in-hospital mortality occurred in 3 patients (21.4%). On a mean follow-up of 9 months, all 11 survivors had noted significant improvement in symptoms without recurrence. CONCLUSIONS: Percutaneous rheolytic thrombectomy using the AngioJet may be a treatment option for patients with massive or submassive PE who may not be eligible for thrombolytic therapy or surgical embolectomy. PMID- 17421020 TI - Mutations and polymorphisms in the human N-acetylglutamate synthase (NAGS) gene. AB - N-acetylglutamate synthase (NAGS) deficiency, an autosomal recessive disorder, is the last urea cycle disorder for which molecular testing became available. This is the first comprehensive report of 21 mutations that cause NAGS deficiency and of commonly found polymorphisms in the NAGS gene. Five mutations are reported here for the first time. A total of 10 disease-causing mutations are associated with acute neonatal hyperammonemia; the remaining mutations were found in patients with late onset disease. Residual enzymatic activities are included in this report and the deleterious effects of eight mutations were confirmed by expression studies. Mutations in the NAGS gene are distributed throughout its reading frame. No mutations have been found in exon 1, which encodes for the putative mitochondrial targeting signal and variable segment of NAGS. Three polymorphisms have been found. Early, accurate, and specific diagnosis of NAGS deficiency is critical since this condition can be successfully treated with N carbamylglutamate (NCG, Carbaglu; Orphan Europe). Treatment with NCG should be initiated as soon as a patient is suspected of having NAGS deficiency. Molecular testing represents the most reliable method of diagnosis. PMID- 17421021 TI - Severe colchicine-induced bilateral optico-neuromyopathy. PMID- 17421023 TI - Access site hematoma requiring blood transfusion predicts mortality in patients undergoing percutaneous coronary intervention: data from the National Heart, Lung, and Blood Institute Dynamic Registry. AB - OBJECTIVE: To determine both the etiology of and outcomes associated with access site hematoma requiring transfusion (HRT) in patients undergoing percutaneous coronary intervention (PCI). BACKGROUND: Access site hematoma in the setting of PCI is the most frequent periprocedural complication (2-12%). Antiplatelet and antithrombin therapy is designed to lower the incidence of adverse ischemic events while maintaining an acceptable rate of hemorrhagic complications. METHODS: This was a prospective, multi-center, cohort study of consecutive patients undergoing PCI during 3 NHLBI Dynamic Registry recruitment waves (1997 2002). The primary endpoints included the incidence of HRT, in-hospital death, and death at 1-year. RESULTS: The incidence of HRT was 1.8% and femoral access was common. Older age, lower BMI, female sex, concomitant renal, cerebrovascular, peripheral vascular, and pulmonary disease were significantly associated with HRT. Glycoprotein IIb/IIIa inhibitors, thrombolytic therapy, and postprocedure heparin were more commonly used in HRT patients, but there was no difference in thienopiridiene use. Attempted lesions in patients developing HRT were more often calcified, thrombotic, located in an ostial location, or class B2 or C. In hospital mortality and 1-year death rate was 9 and 4.5 times higher in HRT patients respectively. Following adjustment, HRT remained independently associated with in-hospital mortality (OR 3.59, 95% CI 1.66-7.77) and 1-year death (hazard ratio [HR] 1.65, 95% CI 1.01-2.70, P = 0.048). Independent predictors of HRT included age, female sex, IIb/IIIa inhibitors, thrombolytic agents, and concomitant conditions. CONCLUSIONS: Access site complications, especially HRT, remain a very important predictor of adverse procedural success and patient outcome. PMID- 17421024 TI - Influence of age on BDNF modulation of hippocampal synaptic transmission: interplay with adenosine A2A receptors. AB - We previously reported that adenosine, through A(2A) receptor activation, potentiates synaptic actions of brain-derived neurotrophic factor (BDNF) in the hippocampus of infant (3-4 weeks) rats. Since A(2A)-receptor-mediated actions are more evident in old than in young rats and since the therapeutic potential for BDNF-based strategies is greater in old subjects, we now evaluated synaptic actions of BDNF and the levels of TrkB receptors and of adenosine A(2A) receptors in the hippocampus of three groups of adult rats: young adults (10-16 weeks), old adults (36-38 weeks), and aged (70-80 weeks), as well as in one group of infant (3-4 weeks) rats. BDNF (20 ng/ml) enhances field excitatory postsynaptic potentials recorded from the hippocampus of young adults and aged rats, an action triggered by adenosine A(2A) receptor activation, since it was blocked by the A(2A) receptor antagonist, ZM 241385. In the other groups of animals BDNF (20 ng/ml) was virtually devoid of action on synaptic transmission. Western blot analysis of receptor density shows decreased amounts of TrkB receptors in old adults and aged rats, whereas A(2A) receptor levels assayed by ligand binding are enhanced in the hippocampus of old adults and aged rats. It is concluded that age related changes in the density of TrkB receptors and of adenosine A(2A) receptors may be responsible for a nonmonotonous variation of BDNF actions on synaptic transmission in the hippocampus. PMID- 17421025 TI - Limits of propidium iodide as a cell viability indicator for environmental bacteria. AB - BACKGROUND: Viability measurements of individual bacteria are applied in various scopes of research and industry using approaches where propidium iodide (PI) serves as dead cell indicator. The reliability of PI uptake as a cell viability indicator for dead (PI permeable) and viable (PI impermeable) bacteria was tested using two soil bacteria, the gram(-) Sphingomonas sp. LB126 and the gram(+) Mycobacterium frederiksbergense LB501T. METHODS: Bacterial proliferation activities observed viaDAPI and Hoechst 33342 staining were linked to the energy charge and the proportion of dead cells as obtained by diOC(6) (3)-staining and PI-uptake, respectively. Calibration and verification experiments were performed using batch cultures grown on different substrates. RESULTS: PI uptake depended on the physiological state of the bacterial cells. Unexpectedly, up to 40% of both strains were stained by PI during early exponential growth on glucose when compared to 2-5% of cells in the early stationary phase of growth. CONCLUSIONS: The results question the utility of PI as a universal indicator for the viability of (environmental) bacteria. It rather appears that in addition to nonviable cells, PI also stains growing cells of Sphingomonas sp. and M. frederiksbergense during a short period of their life cycle. PMID- 17421026 TI - Re-evaluating acridine orange for rapid flow cytometric enumeration of parasitemia in malaria-infected rodents. AB - Methods facilitating research in malaria are of pivotal relevance. Flow cytometry offers the possibility of rapid enumeration of parasitemia. It relies on staining the parasite DNA to distinguish between infected and non-infected red blood cell (RBC) populations. Unfortunately, in rodents abundant reticulocyte RNA interferes with the application of the method. This results in time-consuming sample preparation protocols that offer no clear advantage over microscopic counting. We re-evaluated the use of the DNA/RNA discriminating vital fluorochrome acridine orange (AO) for rapid flow cytometric enumeration of parasitemia in rodents. Whole blood from rodents infected with Plasmodium berghei and Plasmodium yoelii was stained with AO and analyzed by flow cytometer. A newly developed two-channel (FL1/FL3) detection method was compared with conventional one-channel (FL1) detection and microscopic counting. The new AO two-channel detection method clearly discriminated between infected and non-infected RBC populations. It showed to be linear above parasitemias of 0.3%. Sample processing time amounted to approximately 5 min. It is shown that AO can be used for rapid, precise, and accurate enumeration of parasitemia in rodents. Due to its ease of handling the method might find widespread application in malaria research. PMID- 17421027 TI - The human immunodeficiency virus epidemic: a race against time for millions and the role of flow cytometry. A Caribbean and resource-constrained country perspective. AB - There is a race against time for millions in the world today. Both the technology and the manpower are currently available to deliver the services that are required to meet the needs of the 40 million people currently living with HIV/AIDS, but at what price? The reality is therefore that we are a long shot away from this realization. What are the facts and why have we not achieved even the simplest deadlines set by the World Health Organization (WHO)? Are these objectives realistic? What role does hard science have to play in the search for cost-effective solutions and futuristic effective options? To stem this unrelenting epidemic and convert the natural history of the disorder in those already living with the virus into one of chronicity, rather than one characterized by a dehumanizing and stigmatized death, requires a global commitment at all levels. This discussion examines the reality and offers a snapshot of capacity and experiences in the developing world. Crucially, it looks at the immediate and long term role of flow cytometry in the expanded care and treatment programs for developing nations. PMID- 17421028 TI - Joseph Jastrow, the psychology of deception, and the racial economy of observation. AB - This article reconstructs the recurring themes in the career of Joseph Jastrow, both inside and outside the laboratory. His psychology of deception provides the bridge between his experimental and popular pursuits. Furthermore, Jastrow's career illustrates the complex ways in which scientific psychology and pragmatist philosophy operated within the constraints of a moral economy deeply marked by notions of "race." Psychological investigations of deception were grafted onto two of the human sciences' leading tools: the evolutionary narrative and the statistical analysis of populations. Such associations abetted the racialization of the acts of deceiving and being deceived. These connections also were used to craft moral lessons about how individuals ought to behave in relationship to the aggregate population and natural selection's endowment. PMID- 17421030 TI - Unsupported allegations about a link between Milgram and the CIA: tortured reasoning in A Question of Torture. PMID- 17421031 TI - A differential paradox: the controversy surrounding the Scottish mental surveys of intelligence and family size. AB - In 1947, the Scottish Council for Research in Education and the Population Investigation Committee conducted a survey of Scottish schoolchildren, exploring the relations between tested intelligence and fertility. The survey was not only significant for its size, measuring the IQ of all 11-year-olds at school on the day of testing, some 80,805 children, but also because it was a repeat survey. Its purpose was to establish whether the intelligence of the population had declined because of the negative correlation between IQ and family size. The paper will explore how the impetus for the 1947 survey came from attempts to revive the fortunes of the eugenics movement, based upon the interdisciplinary study of population. While most expected the study to provide evidence of a decline in intelligence, it revealed an increase. This was in spite of a continuing process of differential fertility. This paper will explore the influence of these results, described as a "paradox," upon the future development of the eugenics movement and the sciences of population. While for many, the results were seen to have completely, and thankfully, undermined eugenic fears of degeneration, the supposed "resolution" of the paradox in 1962 provided the basis of a meritocratic and optimistic "new eugenics" that sought to reunite social and biological scientists concerned with human betterment in Britain and the United States. PMID- 17421032 TI - Local institutionalization, discontinuity, and German textbooks of psychology, 1816-1854. AB - In the context of social and intellectual developments and the changing role of German universities in the first half of the nineteenth century, which led to the local institutionalization of the discipline of psychology at German universities, the structure and content of textbooks of psychology are discussed. Textbooks in the first half of the nineteenth century had a pedagogical function in training teachers, in socializing students into the field, and in providing students and readers with knowledge about the subject matter, methodology, and topics of psychology. The textbooks, representative of influence, philosophical psychological orientations, and different decades in the first half of the nineteenth century, are reconstructed with regard to the definition of psychology, the ways of studying the soul, and how to conceptually organize the field. The textbooks by Herbart, Beneke, and Waitz, which were written within a natural-scientific programmatic vision for psychology, are contrasted with the traditional philosophically intended textbooks of Reinhold, Mussmann, George, and Schilling. Fischhaber's textbook for Gymnasien is summarized. Issues regarding the continuity of psychology are discussed, and discontinuous developments in the history of German psychology are identified. PMID- 17421034 TI - Alfred McCoy, Hebb, the CIA and torture. PMID- 17421036 TI - Contexts and experimentalism in the psychology of Gabriele Buccola (1875-1885). AB - Gabriele Buccola, since his untimely death, often has been mentioned as the first Italian psychologist who developed a strict program of laboratory research. Buccola, a Sicilian of Albanian ancestry, is a "case" in the history of Italian psychology. A self-taught positivist, he established a relation with the major representatives of the European positivism. Kraepelin mentions him as one of the precursors of his project of applying experimental psychology to psychopathology. Buccola actually carried out research on the psychological, chemical-biological, and psychopathological "modifiers" of reaction times, following an experimental program dealing mainly with the differential study both of basic and superior psychological processes, with mental hygiene ends. Historians of psychology agree in considering Buccola the first Italian laboratory psychologist to plan a program of research that was close to European psychological experimentalism. The present article, starting from an outline of Buccola's role in the rising Italian scientific psychology, recontextualizes his experimentalism in an international sphere. This operation, which is carried out through a careful survey of Buccola's entire production-both theoretical and more properly scientific-is based on the search of the Darwinian, Spencerian, and Haeckelian evolutionist themes emerging from Buccola's program of research-a program that was influenced by the variegated European experimental panorama and characterized by the vision of science as a knowledge capable of transforming the nature of man and of society. PMID- 17421038 TI - Birth season, respiratory viruses and asthma in children. PMID- 17421039 TI - A novel method of encapsulating and cultivating adherent mammalian cells within collagen microcarriers. AB - A novel method of preparing collagen microcarriers was developed and used to entrap adherent cells for cell culturing. This new technique involved seeding of cells in micro gel beads comprised of collagen fibrils dispersed in alginate. The gel beads were washed with phosphate buffered saline (PBS) to remove alginate and the resulting microspheres, about 300-500 microm in diameter, contained evenly distributed collagen fibrils which provided a 3D biomimetic environment for cell growth. The applicability of this microencapsulating system was demonstrated by its ability to support the growth of C2C12 myoblast cells. When seeded and cultured within the 3D collagen microcarriers, the population of C2C12 cells entrapped within the microcarriers increased by 1.5 folds in 7 days after inoculation. This encapsulation technique is potentially useful for culturing cells and especially useful for adherent cells that require a 3D fibrillar collagen environment. PMID- 17421040 TI - Hydrogen sulfide production from elemental sulfur by Desulfovibrio desulfuricans in an anaerobic bioreactor. AB - Feasibility of elemental sulfur reduction by Desulfovibrio desulfuricans in anaerobic conditions in a stirred reactor was studied. Hydrogen was used as energy source, whereas the carbonated species were bicarbonate and yeast extract. Attention was paid to reactor engineering aspects, biofilm formation on the sulfur surface, hydrogen sulfide formation rate and kinetics limitations of the sulfur reduction. D. desulfuricans formed stable biofilms on the sulfur surface. It was found that active sulfur surface availability limits the reaction rate. The reaction rate was first order with respect to sulfur and hydrogen velocity had no effect in the reaction rate for the range 8.2 x 10(-2) to 4.1 x 10(-1) Nm(3) m(-2) min(-1). At a superficial gas velocity (u(G)) = 3.1 x 10(-2) Nm(3) m( 2) min(-1), H(2)S(g) production rate decreased due to a deficient H(2)S stripping. A maximum H(2)S(g) production rate of 2.1 g H(2)S L(-1) d(-1) was achieved during 5 days with an initial sulfur density of 4.7% (w/v). PMID- 17421041 TI - Tracer measurements reveal experimental evidence of biofilm consolidation. AB - The ability to simultaneously measure both biofilm thickness and the mass transfer coefficient of an inert tracer through it provides a powerful method to study biofilm development. In this communication previously published data has been collated to interpret global trends in biofilm structure during the transition towards steady-state. It appears that sudden changes in biofilm structure (directly related to the rate of change of biofilm mass transfer resistance) may occur following transitions in rate of biomass production. These observations are consistent with the concept of consolidation, recently introduced into spatially structured biofilm mathematical models to account for structural realignment of the biofilm under dynamic conditions. PMID- 17421042 TI - On the use of the T-REx tetracycline-inducible gene expression system in vivo. AB - Components of the commercially available T-REx system were used to create two types of transgenic mice. The first contained the tetracycline-repressor transgene under the control of the CMV promoter/enhancer; the second type contained a green fluorescent protein (GFP) reporter transgene under the control of the CMV promoter/enhancer with a tetracycline repressor operator sequence. Transgene expression was unpredictable in animals containing the individual transgenes. Animals with the reporter transgene expressed GFP in only some tissues (e.g., pancreas, kidney), and one line of reporter transgenic animals developed kidney disease, presumably due to expression of the transgene. The two types of transgenic animals were crossbred to produce double-transgenic animals with the object of regulating the expression of the reporter in vivo. When a similar double-transgenic system was constructed in cultured cells, the repressor protein suppressed the transcription of the reporter transgene. The presence of the repressor in double-transgenic animals had no effect on the expression of the reporter; double transgenic animals developed the same kidney disease that was seen in singly transgenic mice with the reporter. Our results indicate that transgenes under the control of the CMV promoter in the T-REx system express somewhat unpredictably and in only a limited number of tissues, making the use of this system for the development of in vivo models problematical. PMID- 17421043 TI - Effect of doxycycline-regulated protein disulfide isomerase expression on the specific productivity of recombinant CHO cells: thrombopoietin and antibody. AB - Protein disulfide isomerase (PDI), one of the ER-resident molecular chaperones, forms and isomerizes disulfide bonds. This study attempts to investigate the effect of PDI expression level on specific productivity (q) of recombinant Chinese hamster ovary (rCHO) cells producing thrombopoietin (TPO) and antibody (Ab). To regulate the PDI expression level, the Tet-Off system was introduced in TPO and Ab producing CHO cells, and stable Tet-Off cells (TPO-Tet-Off and Ab-Tet Off) were screened using the luciferase assay. The doxycycline-regulated PDI expression system in Tet-Off rCHO cells (Tet-TPO-PDI and Tet-Ab-PDI) was established by the cotransfection of pTRE-PDI and pTK-Hyg expression vector into TPO-Tet-Off and Ab-Tet-Off cells, respectively. Subsequent screening was done by Western blot analysis of PDI and an enzyme-linked immunosorbent assay of the secreted TPO and antibody. We cultured two Tet-TPO-PDI and two Tet-Ab-PDI clones, and all these clones showed an average of 2.5-fold increase in PDI expression when compared to the basal level. In both these cell lines the PDI expression was tightly controlled by various concentrations of doxycycline. The q of TPO (q(TPO)) was unaffected but that of antibody producing cells was increased by 15 27% due to the PDI expression level. PMID- 17421044 TI - Optimal fed-batch cultivation when mass transfer becomes limiting. AB - In the design of an aerobic fed-batch process to produce, for example, a pharmaceutical protein, the volumetric production rate will eventually become limited by mass transfer when the biomass concentration exceeds a certain upper limit x*. It appears to be common practice to switch from exponential feed of substrate to a constant feed rate when x* is reached. This is done to avoid oxygen starvation with a potential risk of undesired stress responses. But with a constant feed rate the carbon source (glucose) concentration may decrease to a low level with a resulting loss of viability and an undesired production of endotoxins. It is shown that an exponential feeding strategy may be continued, but with a smaller exponent than the one used before oxygen limitation occurs. This will diminish the potential detrimental effects on the culture due to low glucose concentration, and the total time to reach a given final biomass concentration will be reduced. PMID- 17421045 TI - Optimal thermotolerance of Bifidobacterium bifidum in gellan-alginate microparticles. AB - The purpose of this research was to encapsulate Bifidobacterium bifidum using gellan, sodium alginate and prebiotics as coating materials, and to maximize the thermotolerance of the probiotics with an optimal combination of the coating materials. The optimal ratio of the coating materials for the microparticles under heat treatments (75 degrees C, 1 min) was obtained by using the response surface method and the sequential quadratic programming technique. Optimization results indicated that 2% sodium alginate mixed with 1% gellan gum as coating materials would produce the highest thermotolerance in terms of B. bifidum count. The verification experiment yielded a result close to the predicted values, with no significant difference (P > 0.05). The results of heat treatments also demonstrated that the addition of gellan gum in the walls of probiotic microcapsules provided improved protection for B. bifidum. These probiotic counts remained at 10(5)-10(6) CFU/g for the microcapsules stored for 2 months, then treated in heat and in simulated gastric fluid. PMID- 17421046 TI - Structure and shear strength of microbial biofilms as determined with confocal laser scanning microscopy and fluid dynamic gauging using a novel rotating disc biofilm reactor. AB - The cohesive strength of microbial biofilms cultivated on a rotating disc has been measured using fluid dynamic gauging (FDG). The thickness of heterotrophic mixed culture biofilms was found to depend on substrate concentration and shear force at the biofilm surface during the cultivation. For high substrate concentrations and low shear forces the biofilm thickness increased to several 100 microm within 7 days. Low substrate concentration and higher shear forces yielded thin biofilms of about 100 microm thickness. Independent from cultivation conditions and thickness of the biofilms their cohesive strength ranged between 6.0 and 7.7 N m(-2). The ratio between cohesive strength measured with FDG and shear forces applied during biofilm cultivation have ranged from 200 to 1,100. Higher concentrations of iron in the cultivation media has a positive effect on the stability of the biofilms cultivated. By using the CLSM technique a stable base biofilm with a high amount of stained EPS glycoconjugates could be visualized after gauging. The thickness of the base biofilm was about 100 microm for all biofilms cultivated and was not removable under the applied shear conditions used during FDG. PMID- 17421047 TI - Functional genome-wide analysis of antibody producing NS0 cell line cultivated at different temperatures. AB - Lowering culture temperature has been reported as a significant factor in the improvement of mammalian cell productivity. To determine the physiological changes which take place at different temperature cultivations, an NS0 cell line producing human-mouse chimeric antibody was cultured at 22, 34 and 37 degrees C. Various cellular parameters such as viability, productivity, metabolism, apoptosis and cell cycle were studied and notable changes were shown to be accompanied by changes in metabolic rates. Reduction of the temperature to 22 degrees C resulted in a decrease in the growth rate, inhibition of antibody production, arrest of cell cycle in G2 phase and delay in apoptosis. A slight increase in antibody production was observed at 34 degrees C due to the increase of growth rate and prolonged stationary phase. To better understand and explore the mechanisms underpinning these biological alterations and to identify the genes involved in the genetic reprogramming, genome-wide analyses were performed using GeneChip Mouse Genome arrays. The examination of differential gene expression induced by temperature reduction demonstrated a specific pattern of gene expression in NS0 cells in response to temperature stress. The effect of temperature on transcription induced changes within a wide range of genes involved in metabolic and signalling pathways. Most deregulated genes involved in essential metabolic pathways (i.e. glycolysis/gluconeogenesis, pentose phosphate pathway and inositol metabolism) were repressed in cells cultured at 22 degrees C. By combining gene expression and physiological changes at different temperatures it was possible to provide greater understanding of cell response to hypothermic conditions. PMID- 17421048 TI - Identification of discontinuous antigenic determinants on proteins based on shape complementarities. AB - Diverse procedures for identifying antigenic determinants on proteins have been developed, including experimental as well as computational approaches. However, most of these techniques focus on continuous epitopes, whereas fast and reliable identification and verification of discontinuous epitopes remains barely amenable. In this paper, we describe a computational workflow for the detection of discontinuous epitopes on proteins. The workflow uses a given protein 3D structure as input, and combines a per residue solvent accessibility constraint with epitope to paratope shape complementarity measures and binding energies for assigning antigenic determinants in the conformational context. We have developed the procedure on a given set of 26 antigen-antibody complexes with a known structure, and have further expanded the available paratope shapes by generating a virtual paratope library in order to improve the screening for candidate residues constituting discontinuous epitopes. Applying the workflow on the 26 given antigens with known discontinuous epitopes resulted in the correct identification of the spatial proximity of 12 antigen-antibody interaction sites. Combining solvent accessibility, shape complementarity and binding energies towards the identification of discontinuous epitopes clearly outperforms approaches solely considering accessibility and residue distance constraints. PMID- 17421049 TI - QSAR of multiple mutated antibodies. AB - The aim of this study was to develop predictive quantitative structure-activity relationship (QSAR) modeling for antibody-peptide interactions. A small single chain antibody library was designed and manufactured around the murine anti-p24 (HIV-1) monoclonal antibody CB4-1 by use of statistical molecular design (SMD) principles and site directed mutagenesis, and its affinity for a p24 derived antigen was determined by fluorescence polarization. A satisfactory QSAR model (Q(2) = 0.74, R(2) = 0.88) was derived by correlating the affinity data to physicochemical property scales of the amino acids varied in the library. The model explains most of the antibody-antigen interactions of the studied set, and provides insights into the molecular mechanism involved in antigen binding. PMID- 17421050 TI - Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: the case for conserving natural enemies. AB - Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies. PMID- 17421051 TI - Insecticide toxicity and synergism by enzyme inhibitors in 18 species of pest insect and natural enemies in crucifer vegetable crops. AB - The toxicities of three enzyme inhibitors and their synergistic effects on four insecticides were studied by using the dry film method on field populations of 18 species of insects collected in Jianxin and Shanjie, China, from 2003 to 2005. Meanwhile, the inhibitory effects of these enzyme inhibitors on the activities of acetylcholinesterases (AChE), carboxyesterases (CarE) and glutathione-S transferases (GST), in vivo, were also studied. In general, triphenyl phosphate (TPP) and diethyl maleate (DEM) showed low toxicities to six herbivorous pest insects, four ladybirds and eight parasitoids. Piperonyl butoxide (PB) exhibited low toxicities to the herbivorous pest insects and ladybirds, but high toxicities to the eight parasitoids. The tolerance to the insecticides in 11 pest insects and natural enemies was mainly associated with the tolerance to PB. PB showed the highest synergism on methamidophos, fenvalerate, fipronil and avermectin in nine species of pest insects and natural enemies. In general, TPP and DEM showed significant synergisms to these four insecticides in four parasitoid species. However, in contrast to their effects on the parasitoids, the synergistic effects of TPP and DEM on the four insecticides by TPP and DEM against four pest insects and one ladybird varied depending on the insect species and enzyme inhibitor. Activity of AChE, CarE or GST could be strongly inhibited, in vivo, by PB, TPP or DEM, depending on the insect species and enzyme inhibitors. From the results obtained in this study, mixed-function oxidase (MFO) was thought to play the most critical role in insect tolerances to the tested insecticides in the field. Low competition existed in the evolution of insecticide resistance in the field populations of parasitoids, as compared with herbivorous pest insects and ladybirds. Possible causes of the high synergistic effects of PB on the four classes of insecticides, based on multiattack on the activity of CarE, GST or AChE in the insect species, are also discussed. PMID- 17421052 TI - Can Batesian mimicry help plants to deter herbivores? AB - Several authors have suggested that edible plants could avoid herbivory by mimicking olfactory cues of toxic plants. However, very few studies have been carried out to test this hypothesis. The aims of the present study were to identify the volatiles of three clover species and to test whether a species lacking chemical defences, such as red clover, could avoid being grazed by rabbits by mimicking the volatiles of the cyanogenic white clover. Two main volatiles were identified in all three clover species, and a further two volatiles were present in white clover only. Rabbits presented with a choice between white clover, red clover and red clover sprayed with white clover extract ate significantly more red clover than white or white-flavoured red clover. The results suggest that the volatiles of toxic plants could be used and exploited as a source of natural, safe and effective repellents to control the impact of pest herbivores on plants. PMID- 17421053 TI - Effect of Bt cotton expressing Cry1Ac and Cry2Ab, non-Bt cotton and starvation on survival and development of Trichoplusia ni (Lepidoptera: Noctuidae). AB - Effects of Bollgard II cotton containing two Bacillus thuringensis var. kurstaki Berliner (Bt) toxin proteins (Cry1Ac and Cry2Ab), non-Bt cotton (DPL 491) and starvation on survival and development of cabbage looper, Trichoplusia ni (Hubner), were determined in the laboratory. Larvae of the first four larval instars died when they fed on the terminal leaves of Bt cotton plants at 50 days after planting (DAP). However, 51.3% of fifth instars that fed on 50 DAP Bt cotton leaves pupated, and 87.1% of the pupae successfully developed into adults. Of the unfed fifth instars (starved), 55.6% pupated and 88.1% of the pupae emerged. Pupae that developed from larvae fed on Bt cotton leaves and unfed were significantly smaller, being 89.7 and 73.2% of the weight of the pupae that developed from larvae fed on non-Bt cotton leaves. Leaves of 120 DAP Bt cotton were less toxic to T. ni larvae. When the first instars continuously fed on 120 DAP Bt cotton leaves, 75.9, 60.6, 56.4 and 38.4% of larvae survived to second, third, fourth and fifth instars respectively, and 20.9% pupated and 17.9% successfully became adults. However, it took the surviving first instars 37.1 days to become adults, which was 7.2 and 8.9 days longer than those fed on 50 and 120 DAP non-Bt cottons respectively. Pupae that developed from larvae that fed on 120 DAP Bt cotton leaves were only 50.9 and 52.6% of the weight of those developed from larvae that fed on 50 and 120 DAP non-Bt cotton respectively. Non Bt cotton, both 50 and 120 DAP, did not exhibit significant effects on larval survival and development, except that the pupae in the 50 DAP non-Bt cotton treatments developed over a significantly longer time than those in the 120 DAP non-Bt cotton treatment. PMID- 17421054 TI - Repellence of the red bud borer Resseliella oculiperda from grafted apple trees by impregnation of rubber budding strips with essential oils. AB - The red bud borer Resseliella oculiperda (Rubs.) is a pest insect of apple trees when rootstocks are grafted with scion buds by 'shield budding'. The female midges are attracted to the wounds of the grafted buds where they lay their eggs. The larvae feed on the cambium and destroy the buds completely or partially, leading to bad union of the buds with the rootstocks. Budding strips are used very often by growers to bind scion buds to rootstocks. These strips cannot prevent midges from reaching the damaged tissue. Chemical treatments applied to the grafts and other types of strip do not provide better protection against the pest and may cause other risks for growers. In orchard experiments in 2000 and 2001, the authors evaluated the repellent action provided by three essential oils and five compounds of plant origin against the midges by impregnating budding strips with them. The essential oils of lavender, Lavandula angustifolia (P. Mill.), and alpha-terpineol decreased the infestation of buds by more than 95 and 80% respectively. The other potential repellents tested [the essential oil of Juniperus virginiana (L.), citronellal, the essential oil of Cinnamomum camphora (L.) J. Presl, R-carvone, linalool and R-fenchone] decreased infestation by 67, 66, 51, 45, 37 and 25% respectively. The formulation and commercial development of budding strips impregnated with lavender oil is discussed. PMID- 17421055 TI - Improved short-term engraftment of lentivirally versus gammaretrovirally transduced allogeneic canine repopulating cells. AB - Gammaretroviral vectors require cell division for efficient transduction. Thus, extended cell culture times are necessary for efficient transduction with gammaretroviral vectors, which in turn can lead to stem cell loss and impaired engraftment. Lentiviral vectors transduce nondividing cells and are therefore able to transduce stem cells in short transduction protocols. Here, we compared the short-term engraftment of lentivirally and gammaretrovirally transduced canine allogeneic DLA-matched littermate cells. A reduced conditioning regimen of 400 cGy total body irradiation was used in preparation for clinical studies. Two dogs received a graft of gammaretrovirally transduced CD34-selected cells. CD34(+) cells were prestimulated for 30 h and then exposed twice to concentrated RD114 pseudotype vector. Three dogs received lentivirally transduced CD34 selected cells. Cells were transduced overnight with concentrated VSV-G pseudotype lentiviral vector. The animals in the lentiviral group showed a significantly faster granulocyte recovery. VNTR analysis 40-50 days after transplantation revealed higher donor chimerism for the lentiviral group compared to the retroviral group. These data suggest that short lentiviral transduction protocols may be superior to extended gammaretroviral transduction protocols with respect to engraftment potential of transduced CD34(+) hematopoietic repopulating cells. PMID- 17421056 TI - Mechanisms by which Astragalus membranaceus injection regulates hematopoiesis in myelosuppressed mice. AB - The aim of this study was to investigate the mechanism underlying the effects of Astragalus membranaceus injection (AMI) on myelopoiesis in myelosuppressed mice. At 72 h after cyclophosphamide injection (250 mg/kg), the mice were administered AMI (500 mg/kg) intraperitoneally for 6 consecutive days or an equivalent volume of saline as a control. Murine colony-forming unit-fibroblast (CFU-F) formation, production of IL-6 and GM-CSF by bone marrow stromal cells (BMSC), and bcl-2 protein and mRNA expression in BMSC were measured by CFU-F assay, ELISA, immunohistochemistry and in situ hybridization. The results indicated that AMI improved the hematopoietic microenvironment by enhancing the BMSC survival and proliferation of CFU-F, production of IL-6 as well as GM-CSF by BMSC and bcl-2 protein and mRNA expression in BMSC, which promoted myelopoiesis. The data may provide a mechanistic basis for applying this ancient Chinese herb to promote hematopoiesis as an efficacious adjuvant therapy against myelosuppression induced by anti-cancer therapy. PMID- 17421057 TI - Reproductive toxicity of lapachol in adult male Wistar rats submitted to short term treatment. AB - Lapachol is a therapeutic naphthoquinone, but little is known about its general and reproductive toxicity. In female rats, a high incidence of resorptions and fetal mortality has been reported. This work analyses the effect of the short term administration of lapachol on vital and reproductive organs, and sperm production in Wistar rats. Adult animals were treated with 1 mL of lapachol hydroalcohol solution (100 mg/kg of body weight) for 5 days and killed 3 (T1) and 14 days (T2) after the end of treatment. Body and organ weights and sperm production were evaluated. The administration of lapachol significantly reduced the weight of the seminal vesicle (T1 animals). No significant alteration of gamete production, body weight and the weight of the other organs analysed were detected. The results suggest a reproductive toxicity effect of lapachol, indicating the seminal vesicle as a possible target organ. PMID- 17421058 TI - Antibacterial activity of two canthin-6-one alkaloids from Allium neapolitanum. AB - The emergence of multidrug-resistant strains of many human pathogens has led to an urgent need for the discovery and development of new antimicrobial agents. As part of an ongoing investigation into the antibacterial properties of the Alliaceae, the isolation of 1 (canthin-6-one), 2 (8-hydroxy-canthin-6-one) and 3 (5(zeta)-hydroxy-octadeca-6(E)-8(Z)-dienioc acid)) from A. neapolitanum, a perennial bulbous herb found in open pastures of the Mediterranean is reported. Compounds 1 and 2 were isolated by Sephadex LH-20 from fractions exhibiting a positive reaction with Dragendorff's reagent on TLC, compound 3 was isolated after HPLC purification of Sephadex fractions. Structures were elucidated by extensive 1D and 2D NMR experiments and are in accordance with published data, however, the 13C NMR data for compound 2 and the 1H and 13C NMR data for compound 3 are reported here for the first time. Canthin-6-one alkaloids are well-known constituents of the Simaroubaceae and Rutaceae, and display a wide range of biological activities. These metabolites are reported as constituents of the Alliaceae here for the first time, and displayed minimum inhibitory concentrations (MICs) in the range 8-32 microg/mL against a panel of fast-growing Mycobacterium species and 8-64 microg/mL against multidrug-resistant (MDR) and methicillin-resistant (MRSA) strains of Staphylococcus aureus. Compound 3 displayed antimycobacterial activity in the range of 16-32 microg/mL. PMID- 17421059 TI - Bio-guided isolation of antioxidants from the stems of Dendrobium aurantiacum var. denneanum. AB - Bio-guided fractionation of the stems of Dendrobium aurantiacum var. denneanum using a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay, led to the isolation of three 2-glucosyloxycinnamic acid derivates, namely, cis melilotoside, trans-melilotoside and dihydromelilotoside, respectively. Their structures were elucidated through the analysis of uni- and bi-dimensional NMR, UV, IR and MS data. All these three compounds were first reported from the genus Dendrobium and exhibited potent antioxidant activities. PMID- 17421060 TI - Directing endothelial differentiation of human embryonic stem cells via transduction with an adenoviral vector expressing the VEGF(165) gene. AB - Endothelial progenitors derived from human embryonic stem cells (hESCs) hold much promise in clinical therapy. Conventionally, lineage-specific differentiation of hESCs is achieved through supplementation of various cytokines and chemical factors within the culture milieu. Nevertheless, this is a highly inefficient approach that is often limited by poor replicability. An alternative is through genetic modulation with recombinant DNA. Hence, this study investigated whether transduction of hESCs with an adenoviral vector expressing the human VEGF(165) gene (Ad-hVEGF(165)) can enhance endothelial-lineage differentiation. The hESCs were induced to form embryoid bodies (EBs) by culturing them within low attachment plates for 7 days, and were subsequently trypsinized into single cells, prior to transduction with Ad-hVEGF(165). Optimal transduction efficiency with high cell viability was achieved by 4-h exposure of the differentiating hESCs to viral particles at a ratio of 1 : 500 for three consecutive days. ELISA results showed that Ad-hVEGF(165)-transduced cells secreted human vascular endothelial growth factor (hVEGF) for more than 30 days post-transduction, peaking on day 8, and the conditioned medium from the transduced cells stimulated extensive proliferation of HUVEC. Real-time PCR analysis showed positive upregulation of VEGF, Ang-1, Flt-1, Tie-2, CD34, CD31, CD133 and Flk-1 gene expression in Ad-hVEGF(165)-transduced cells. Additionally, flow cytometric analysis of CD133 cell surface marker revealed an approximately 5-fold increase in CD133 marker expression in Ad-hVEGF(165)-transduced cells compared to the non transduced control. Hence, this study demonstrated that transduction of differentiating hESCs with Ad-hVEGF(165) facilitated expression of the VEGF transgene, which in turn significantly enhanced endothelial differentiation of hESCs. PMID- 17421061 TI - Re: Pathologic guidelines for orthotopic urinary diversion in women with bladder cancer: a review of the literature. PMID- 17421062 TI - Re: Reevaluating occult stress incontinence. PMID- 17421063 TI - Re: A molecular correlate to the Gleason grading system for prostate adenocarcinoma. PMID- 17421064 TI - Re: Metastatic prostate cancer-does treatment of the primary tumor matter? PMID- 17421065 TI - Re: Vaccination of hormone-refractory prostate cancer patients with peptide cocktail-loaded dendritic cells: results of a phase 1 clinical trial. PMID- 17421066 TI - Re: Primary T1G3 bladder cancer: organ preserving approach or immediate cystectomy? PMID- 17421067 TI - Report calls for changes in US global AIDS efforts. PMID- 17421068 TI - Bulgarian doctors protest over crisis in health-care system. PMID- 17421069 TI - [Problems involved in expert opinions on acceleration injuries of the cervical spine]. PMID- 17421070 TI - A useful agent for chemoprevention of hepatocellular carcinoma? AB - Hepatocellular carcinoma (HCC) is a major health problem worldwide, involving more than 620,000 new cases yearly, with chronic HBV and HCV infection being the most common causes. Because most patients are diagnosed in an advanced state which is usually not amenable to curative therapy, prevention should be the main focus for reducing the HCC incidence and its related morbidity and mortality. Strategies of HCC prevention can focus on each single risk factor, with antiviral therapy against chronic hepatitis B and hepatitis C as well as suppression of the progression of common liver diseases being the most important and effective measures. Bicyclol, a drug that can improve liver function and inhibit HBV replication, may be a useful agent for the chemopreventive of HCC, as indicated by a recent study by Zhu et al. Bicyclol can act on the initiation and promotion stages of hepatocarcinogenesis by preventing malignant transformation of hepatic cells. It may also enhance the liver's capacity of detoxification and inhibit DNA mutations. More clinical studies are needed to further demonstrate the efficacy and molecular mechanisms of this agent on HCC chemoprevention. PMID- 17421071 TI - [Do second-opinion centers improve the quality of health care? Statement]. PMID- 17421072 TI - Leaks in the pipeline. PMID- 17421073 TI - Translating science into business. PMID- 17421074 TI - Found in translation. PMID- 17421075 TI - Alternatives in Colorado. PMID- 17421076 TI - The centennial of Cancer Science. PMID- 17421077 TI - Fluorescence lifetimes study of alpha-tocopherol and biological prenylquinols in organic solvents and model membranes. AB - We have found that for biological prenyllipids, such as plastoquinol-9, alpha tocopherol quinol, and alpha-tocopherol, the shortest fluorescence lifetimes were found in aprotic solvents (hexane, ethyl acetate) whereas the longest lifetimes were those of ubiquinonol-10 in these solvents. For all the investigated prenyllipids, fluorescence lifetime in alcohols increased along with an increase in solvent viscosity. In a concentrated hexane solution, the lifetimes of prenylquinols considerably decreased. This contrasts with methanol solutions, which is probably due to the self-association of these compounds in aprotic solvents. We have also found a correlation of the Stokes shift of prenyllipids fluorescence with the orientation polarizability of the solvents. Based on data obtained in organic solvents, measurements of the fluorescence lifetimes of prenyllipids in liposomes allowed an estimation of the relative distance of their fluorescent rings from the liposome membrane surface, and was found to be the shortest for alpha-tocopherol quinol in egg yolk phosphatidylcholine liposomes, and increased in the following order: alpha-tocopherol in dipalmitoyl phosphatidylcholine liposomes < alpha-tocopherol < plastoquinol-9 < ubiquinol-10 in egg-yolk phosphatidylcholine liposomes. PMID- 17421078 TI - Tumor vascular area correlates with photosensitizer uptake: analysis of verteporfin microvascular delivery in the Dunning rat prostate tumor. AB - The parameters that limit supply of photosensitizer to the cancer cells in a solid tumor were systematically analyzed with the use of microvascular transport modeling and histology data from frozen sections. In particular, the vascular permeability transport coefficient and the effective interstitial diffusion coefficient were quantified for Verteporfin-for-Injection delivery of benzoporphyrin derivative (BPD). Orthotopic tumors had higher permeability and diffusion coefficients (Pd = 0.036 microm/s and D = 1.6 microm(2)/s, respectively) as compared to subcutaneously grown tumors (Pd = 0.025 microm/s and D = 0.9 microm2/s, respectively), likely due to the fact that the vessel patterns are more homogeneous orthotopically. In general, large intersubject and intratumor variability exist in the verteporfin concentration, in the range of 25% in plasma concentration and in the range of 20% for tissue concentrations, predominantly due to these microregional variations in transport. However, the average individual uptake of photosensitizer in tumor tissue was only correlated to the total vascular area within the tumor (R2 = 64.1%, P < 0.001). The data are consistent with a view that microregional variation in the vascular permeability and interstitial diffusion rate contribute the spatial heterogeneity observed in verteporfin uptake, but that average supply to the tissue is limited by the total area of perfused blood vessels. This study presents a method to systematically analyze micro-heterogeneity as well as possible methods to increase delivery and homogeneity of photosensitizer within tumor tissue. PMID- 17421079 TI - A new approach for studying fast biological reactions involving nitric oxide: generation of NO using photolabile ruthenium and manganese NO donors. AB - Nitric oxide (NO) is recognized as one of the major players in various biochemical processes, including blood pressure, neurotransmission and immune responses. However, experimental studies involving NO are often limited by difficulties associated with the use of NO gas, including its toxicity and precise control over NO concentration. Moreover, the reactions of NO with biological molecules, which frequently occur on time scales of microseconds or faster, are limited by the millisecond time scale of conventional stopped-flow techniques. Here we present a new approach for studying rapid biological reactions involving NO. The method is based on designed ruthenium and manganese nitrosyls, [Ru(PaPy3)(NO)](BF4)2 and [Mn(PaPy3)(NO)](ClO4) (PaPy3H = N,N-bis(2 pyridylmethyl)amine-N-ethyl-2-pyridine-2-carboxamide), which upon photolysis produce NO on a fast time scale. The kinetics of the binding of the photogenerated NO to reduced cytochrome c oxidase (CcO) and myoglobin (Mb) was investigated using time-resolved optical absorption spectroscopy. The NO was found to bind to reduced CcO with an apparent lifetime of 77 micros using the [Mn(PaPy3)(NO)]+ complex; the corresponding rate is 10-20 times faster than can be detected by conventional stopped-flow methods. Second-order rate constants of approximately 1 x 10(8) M(-1) s(-1) and approximately 3 x 10(7) M(-1) s(-1) were determined for NO binding to reduced CcO and Mb, respectively. The generation of NO by photolysis of these complexes circumvents the rate limitation of stopped flow techniques and offers a novel alternative to study other fast biological reactions involving NO. PMID- 17421080 TI - Commentary: improving quality of life is a morally important goal for gene therapy. PMID- 17421081 TI - [Radioisotope department in Motol--the flow of time]. AB - The radioisotope department in Motol was established on the 1st of June 1957 as the part of The Research Institute of Endocrinology founded by Assoc. Prof. Karel Silink. In the beginning, the department included 20 beds and its main activity consisted of radioiodine treatment of hyperthyroidism. Since 1965 the attention has been paid to the treatment of thyroid cancer, and this tradition continues till this time. The article reviews the main directions of development of this department since the very beginning to present. PMID- 17421082 TI - A case of unilateral hypotension. PMID- 17421083 TI - Adam's apple: be aware of cultural beliefs. PMID- 17421084 TI - Discrepancies in administrative databases: implications for practice and research. PMID- 17421085 TI - Still searching for the magic food. PMID- 17421086 TI - Dear levosimendan, the right ventricle will thank you! PMID- 17421087 TI - Ushering in the era of nuclear terrorism. PMID- 17421088 TI - Protocols, practice, and patients--the case of alcohol withdrawal. PMID- 17421089 TI - Morbidly obese patients with acute respiratory failure: don't reach for the endotracheal tube yet! PMID- 17421090 TI - Debriefing is an effective method for providing feedback and ensuring adherence to best clinical practice by residents in the intensive care unit. PMID- 17421091 TI - Disparities in sepsis: what do we understand? PMID- 17421092 TI - Moving beyond numbers ... the next step forward in improving patient care. PMID- 17421093 TI - Open lung ventilation: waiting for outcome studies? PMID- 17421094 TI - Mortality and tracheotomy. PMID- 17421095 TI - You never know--one of your patients with cancer might surprise you. PMID- 17421096 TI - Eliminating pressure ulcers: do specialty beds or specialty nurses matter more? PMID- 17421097 TI - Septic plasma-induced oxidative stress in endothelial cells: a sensitive bioassay predicting outcome in septic shock? PMID- 17421098 TI - Declining standardized mortality ratios: how we treat or whom we treat? PMID- 17421099 TI - Actin-binding plasma gelsolin: a potential future ally in the fight against sepsis. PMID- 17421100 TI - Sepsis-induced myocardial depression: where is the missing link? PMID- 17421101 TI - PEEP in the morning, PEEP at night. PMID- 17421102 TI - Understanding another acute respiratory distress syndrome. PMID- 17421103 TI - Genetic influences on severe lung injury: how many more genes? PMID- 17421104 TI - Never the tube! Try the mask! PMID- 17421105 TI - Randomized, controlled trials in the emergency setting: a matter of physician patient relationships, responsibility, and trust. PMID- 17421106 TI - Intensive care unit treatment in patients > 65 yrs with a first-day sequential organ failure assessment score > 15 is not futile. PMID- 17421107 TI - Anti-inflammatory activity of albumin. PMID- 17421108 TI - Endotracheal intubation in the intensive care unit: non-anesthesiologists know rapid sequence intubation, but is it accurate in all cases? PMID- 17421109 TI - The future of surgical critical care: a European perspective. PMID- 17421110 TI - Silver-coated endotracheal tubes: is the bactericidal effect time limited? PMID- 17421111 TI - Albumin supplementation and organ function. PMID- 17421112 TI - High-quality enteral nutrition essential for reducing morbidity and mortality in acute care patients. PMID- 17421113 TI - Saving lives in severe sepsis with the help of enteral nutrition. PMID- 17421114 TI - Circulating levels of soluble triggering receptor expressed on myeloid cells (sTREM)-1 in community-acquired pneumonia. PMID- 17421115 TI - What does it mean: appropriate therapy for methicillin-resistant Staphylococcus aureus? PMID- 17421116 TI - The findings of the International Conference on Medical Emergency Teams are biased and misleading. PMID- 17421117 TI - Requirements of the afferent arm of rapid response systems. PMID- 17421118 TI - Early goal-directed therapy of septic shock: we honestly remain skeptical. PMID- 17421120 TI - A New Year's Message from Our President. PMID- 17421119 TI - Selenium in intensive care (SIC) study: the XX files are still unresolved. PMID- 17421121 TI - Nutritional Muscular Dystrophy in Calves. I. PMID- 17421122 TI - Bat Transmitted Paralytic Rabies in Trinidad. PMID- 17421123 TI - Trichobezoar or Hair Ball in a Cat. PMID- 17421124 TI - Identical Fibro-Sarcomata on Two Mute Swans. PMID- 17421125 TI - A Problem in Pigs. PMID- 17421126 TI - The Prospect Before Us. PMID- 17421127 TI - PERSONAL NEWS. PMID- 17421128 TI - NEWS NOTES. PMID- 17421129 TI - Random Notes On Veterinary Education. PMID- 17421130 TI - Bat Transmitted Paralytic Rabies in Trinidad. PMID- 17421131 TI - Variations in the Effectiveness of Commercial Infectious Feline Enteritis Vaccines in Preventing Virus Enteritis of Mink. PMID- 17421132 TI - Displaced Abomasum-A Method of Correction. PMID- 17421133 TI - Radiology-Part I. PMID- 17421134 TI - The Practitioner and the Radiologist. PMID- 17421137 TI - Surgical Correction of a Recto-Vaginal Fistula in a Mare. PMID- 17421136 TI - Fracture and Dislocation of a Femoral Head. PMID- 17421138 TI - Puzzling Symptoms in a Bitch. PMID- 17421140 TI - Les Deux Aspects de la vibriose bovine: Vibriose venerienne et avortement vibrionien. PMID- 17421139 TI - Are Veterinarians "On The Way Out" Again? PMID- 17421142 TI - PERSONAL NEWS. PMID- 17421141 TI - NEWS NOTES. PMID- 17421144 TI - Hip Dysplasia and the Veterinarian. PMID- 17421143 TI - La Recherche-promesse d'avenir. PMID- 17421145 TI - Congenital Interventricular Septal Defect in a Heifer. PMID- 17421146 TI - Algal Poisoning in Beef Cattle. PMID- 17421148 TI - Rumenotomy and Intestinal Anastomosis in a Heifer. PMID- 17421147 TI - A Case of Strongyloides Dermatitis in Canada. PMID- 17421149 TI - Nematodiriasis in Calves. PMID- 17421152 TI - NEWS NOTES. PMID- 17421150 TI - Foreign Body in the Trachea. PMID- 17421153 TI - PERSONAL NEWS. PMID- 17421155 TI - The Veterinarian and Income Tax. PMID- 17421154 TI - La MeDecine Veterinaire devant son destin. PMID- 17421156 TI - Dr Gustave Labelle. PMID- 17421158 TI - Le Veterinaire et son Impot. PMID- 17421157 TI - A Modified Direct Complement-Fixation Test for the Detection of Antibodies for Salmonella Antigens in Turkey Sera. PMID- 17421160 TI - Parasitic Gastro-Enteritis of Sheep. PMID- 17421159 TI - The Development of Resistance to Terramycin by Intestinal Bacteria of Swine. PMID- 17421161 TI - Lightning Stroke. PMID- 17421163 TI - Rumen Overload. PMID- 17421162 TI - Radiology, II. PMID- 17421164 TI - ANIMAL DENTISTRY: I. Diseases of the Teeth and their Supporting Structures. PMID- 17421165 TI - ANIMAL DENTISTRY: II. Dental Flap Operation for Tooth Extraction. PMID- 17421166 TI - An Outbreak of Myoglobinuria in Light Horses. PMID- 17421167 TI - Quintuplet Births in an Angus Cow. PMID- 17421168 TI - PERSONAL NEWS. PMID- 17421169 TI - Blastomycosis in a Dog. PMID- 17421170 TI - NEWS NOTES. PMID- 17421172 TI - Infectious Canine Hepatitis Modified Live Virus Vaccine Produced in Ferret Kidney Cells. PMID- 17421173 TI - Arteriosclerotic Changes in the Visceral Arteries of Sheep. PMID- 17421174 TI - Ketosis in Cattle-a Review. PMID- 17421175 TI - Problems in Dairy Cattle Practice-Remarks. PMID- 17421176 TI - The Modern Treatment of Traumatic Spinal Compressions in the Dog. PMID- 17421177 TI - Cystic Calculus in a Three-Year-Old Gelding. PMID- 17421178 TI - PERSONAL NEWS. PMID- 17421179 TI - Prolonged Local Anesthesia with Alcohol. PMID- 17421180 TI - Listeria monocytogenes from a Nova Scotia Moose. PMID- 17421181 TI - Intramedullary Pinning of Bilateral Fractures of the Shaft of the Ilium. PMID- 17421182 TI - NEWS NOTES. PMID- 17421183 TI - LETTER TO THE EDITOR. PMID- 17421184 TI - Toxic Waterblooms of Blue-Green Algae. AB - Unialgal cultures of several species and strains of blue-green algae, including those most suspected of causing animal deaths, have been grown and found to vary greatly in toxicity. At least four toxic factors have been recognized. One produces fast deaths and is algal in origin. The others produce slow deaths and are bacterial in origin. The fast-death factor (FDF) is an endotoxin that so far has been encountered only with strains of Microcystis aeruginosa Kutz. emend. Elenkin. Its production is genetically and physiologically controlled. An FDF producing strain of M. aeruginosa has been cultured on a large scale and the cells shown to be toxic when administered orally to sheep, calves, and smaller animals. FDF isolated from these cells has been identified as a quite stable cyclic polypeptide having an intraperitoneal LD(50) for white mice of 0.47 mg. per kg. body weight. The slow-death factors may also contribute to the toxicity of waterblooms. It is concluded that a complex of interdepedent variables determines the degree and kind of toxicity that a waterbloom can develop. PMID- 17421186 TI - Changeover from Large to Small Animal Practice. PMID- 17421185 TI - A Discussion of Some Diseases of Swine. PMID- 17421187 TI - How Much Does It Cost? PMID- 17421189 TI - Arthritis in a Bitch Caused by Salmonella heidelberg. PMID- 17421188 TI - Transthoracic Surgery. PMID- 17421190 TI - Resection of the Left Suprapharyngeal Lymph Gland in a Dairy Cow. PMID- 17421191 TI - PERSONAL NEWS. PMID- 17421192 TI - An Unusual Automobile Injury. PMID- 17421193 TI - NEWS NOTES. PMID- 17421194 TI - LETTERS TO THE EDITOR. PMID- 17421195 TI - LETTERS TO THE EDITOR. PMID- 17421196 TI - Some Possible Applications of Atomic Energy in Veterinary Medicine. PMID- 17421197 TI - Salmonella pullorum in Turkeys-Experimental Testing in Natural Infection. PMID- 17421198 TI - Semen Storage in Liquid Nitrogen. PMID- 17421199 TI - Two Outbreaks of Bovine Trichomoniasis in Herds in Southwestern Ontario. PMID- 17421200 TI - Blood Sugar Levels in Normal Cows before and after Treatment with Prednisolone and Dexamethasone. PMID- 17421201 TI - Evolution de l'elevage des animaux a fourrure dans la Province de Quebec. PMID- 17421202 TI - Radiology III. PMID- 17421203 TI - Bone Pinning in a Mature Bull. PMID- 17421205 TI - Posterior Paralysis in a Cocker Spaniel. PMID- 17421204 TI - The Diagnosis, Radiographic Study and Surgical Relief of a Case of Oesophageal Diverticulum. PMID- 17421206 TI - LETTERS TO THE EDITOR. PMID- 17421208 TI - NEWS NOTES. PMID- 17421207 TI - LETTERS TO THE EDITOR. PMID- 17421209 TI - PERSONAL NEWS. PMID- 17421211 TI - Bovine Abortion-An Analysis of 227 Cases. PMID- 17421212 TI - Public Health in our Changing Environment: The Role of the Veterinarian. PMID- 17421213 TI - Some Possible Applications of Atomic Energy in Veterinary Medicine. PMID- 17421214 TI - The Veterinarian's Future in Agricultural Integration. PMID- 17421215 TI - DENTAL CARIES IN DOGS. PMID- 17421216 TI - Surgical Drainage of the Bovine Pericardial Sac. PMID- 17421217 TI - Gangrenous Mastitis Associated with Intravascular Hemolysis in a Jersey Cow Three Weeks Antepartum. PMID- 17421218 TI - An Unusual Case of Masticatory Muscular Atrophy in a Dog. PMID- 17421219 TI - A Case of Acute Pancreatitis in a Dog. PMID- 17421220 TI - Sedatives and Shows. PMID- 17421221 TI - PERSONAL NEWS. PMID- 17421222 TI - LETTERS TO THE EDITOR. PMID- 17421223 TI - NEWS NOTES. PMID- 17421224 TI - TUMOURS OF THE GUMS. PMID- 17421225 TI - The Organization and Work of the Veterinary Services of Great Britain. PMID- 17421227 TI - PERSONAL NEWS. PMID- 17421226 TI - Recent Progress in Disease Eradication in Canada. PMID- 17421228 TI - The Vanishing Scene. PMID- 17421229 TI - Bloat-Why? PMID- 17421230 TI - Achlorhydria in Dogs, WITH Report of a Case Complicated by Avitaminosis C. PMID- 17421232 TI - The Use of Intravenous Catheterization as an Aid to Fluid Therapy. PMID- 17421231 TI - LETTERS TO THE EDITOR: A FULL-TIME EXECUTIVE SECRETARY. PMID- 17421233 TI - Coelosomie Chez un Foetus de Bison (Bison bison). PMID- 17421234 TI - Torsion of the Cecum in Cattle. PMID- 17421236 TI - Tympanites in a Lioness. PMID- 17421235 TI - Anophthalmia or Congenital Absence of the Eyes in a Calf. PMID- 17421237 TI - Nasopharyngeal Polyps in a Kitten. PMID- 17421238 TI - Ringworm in a Siamese Cattery. PMID- 17421239 TI - LETTER TO THE EDITOR: HAEMOLYTIC DISEASE OF NEW-BORN PIGS. PMID- 17421240 TI - NEWS NOTES. PMID- 17421242 TI - Nutritional Muscular Dystrophy in Calves. III Purkinje Fiber Degeneration in Calves with Experimentally Produced Nutritional Muscular Dystrophy. PMID- 17421244 TI - LETTER TO THE EDITOR: Tularemia. PMID- 17421243 TI - A Review of Current Knowledge of the Vesicular Diseases. PMID- 17421245 TI - Diseases of the Brain of Calves. PMID- 17421246 TI - Surgical Repair of Scrotal Hernia in the Bull. PMID- 17421247 TI - Pneumoencephalography in the Dog. PMID- 17421248 TI - Haemolytic Disease of New-Born Pigs. PMID- 17421250 TI - Pityriasis Rosea in a Herd of Swine. PMID- 17421249 TI - Letters To The Editor. PMID- 17421252 TI - NEWS NOTES. PMID- 17421251 TI - Post-Operative Intussusception in the Dog. PMID- 17421253 TI - PERSONAL NEWS. PMID- 17421254 TI - Foot-and-Mouth Disease in the Americas. PMID- 17421255 TI - The Veterinarian and Civil Defence. PMID- 17421256 TI - Diseases of the Brain of Calves (concluded). PMID- 17421257 TI - Bovine Pulmonary Emphysema. PMID- 17421258 TI - Radiology IV. PMID- 17421260 TI - Scrapie and Kuru. PMID- 17421259 TI - Simple Fixation for Distal Epiphyseal Fractures of the Femur and Tibia in the Cat and Dog. PMID- 17421261 TI - Another Veterinary College? PMID- 17421262 TI - Malignant Lymphoma in a Cow. PMID- 17421263 TI - Displaced Abomasum in Cattle. PMID- 17421264 TI - Perineal Hernia in a Bitch. PMID- 17421265 TI - Aortic Valve Incompetence in a Doberman. PMID- 17421266 TI - Another Veterinary College? PMID- 17421267 TI - NEWS NOTES. PMID- 17421268 TI - PERSONAL NEWS. PMID- 17421270 TI - LETTERS TO THE EDITOR. PMID- 17421269 TI - Tranquillizers and Race Horses. PMID- 17421271 TI - JEWISH SLAUGHTER. PMID- 17421273 TI - Common Conditions Encountered in Race Horses. PMID- 17421274 TI - Abdominal Surgery in the Horse. PMID- 17421275 TI - A Veterinarian and Harness Racing. PMID- 17421276 TI - PERSONAL NEWS. PMID- 17421277 TI - Some Problems and Disorders Encountered in Light Horse Practice. PMID- 17421278 TI - Clostridium novyi in Cattle. PMID- 17421279 TI - Harness for Supporting Urine Collecting Bag. PMID- 17421280 TI - Diagnosis and No Visible Lesions in Tuberculosis of Cattle. PMID- 17421281 TI - Sterile Milk as a Semen Diluent. PMID- 17421282 TI - Radiology V. PMID- 17421283 TI - A Discussion of Small Animal House Practice. PMID- 17421284 TI - Tranquilizers in the Treatment of Tetanus in the Horse. PMID- 17421285 TI - Necrotic Vulvitis in a Herd of Dairy Cattle. PMID- 17421287 TI - ETHICS. PMID- 17421286 TI - Idiopathic Thrombocytopenic Purpura in a Dog. PMID- 17421290 TI - New Year's Message from our President. PMID- 17421288 TI - NEWS NOTES. PMID- 17421291 TI - The Place of the Veterinarian in Municipal Milk and Food Inspection. PMID- 17421292 TI - The Veterinarian and Thermonuclear Warfare. PMID- 17421294 TI - Aspergillosis in a Calf. PMID- 17421293 TI - Avian Vibrionic Hepatitis in Alberta. PMID- 17421295 TI - Hepatic Distomatosis and Infectious Canine Hepatitis in Northern Manitoba. PMID- 17421296 TI - Rare Cause for Incontinence in a Bitch. PMID- 17421297 TI - Surgical Correction of Urinary Retention in the Cat. PMID- 17421298 TI - PERSONAL NEWS. PMID- 17421299 TI - NEWS NOTES. PMID- 17421300 TI - Another Veterinary College? PMID- 17421301 TI - LETTERS TO THE EDITOR. PMID- 17421302 TI - The Transaminase Test for Liver Necrosis in Dogs. PMID- 17421303 TI - Toxoplasmosis as a Cause Of Abortion in Ontario Sheep. PMID- 17421304 TI - ANNOTATION: MAST CELLS. PMID- 17421305 TI - Histopathology in a Case of Feline Urolithiasis. PMID- 17421306 TI - Some Notes on the History and Economics of Harness Racing. PMID- 17421307 TI - Some Findings Concerning a Report on the Cultivation of Feline Panleukopenia Virus in Cell Culture. PMID- 17421309 TI - Feeding Stuffs and Pesticide Regulations in Canada. PMID- 17421308 TI - Conservative Treatment of Traumatic Reticulitis. PMID- 17421310 TI - A Remedy Overlooked-the Gene. PMID- 17421312 TI - NEWS NOTES. PMID- 17421311 TI - Intermandibular Abscess in a Cow. PMID- 17421313 TI - The Syndrome of Bovine Pulmonary Emphysema. PMID- 17421315 TI - Ethics and public relations. PMID- 17421317 TI - PERSONAL NEWS. PMID- 17421316 TI - North American Blastomycosis in the Dog: With a Report of Six Canadian Cases. PMID- 17421318 TI - Irradiation of Embryonated Eggs: Preliminary Report. PMID- 17421319 TI - Sodium Salt Poisoning in Swine. PMID- 17421320 TI - Torsion of an Ectopic Testicle Simulating a Foreign Body in the Intestine. PMID- 17421321 TI - Extensive Lipoma in a Dachshund. PMID- 17421322 TI - NEWS NOTES. PMID- 17421323 TI - The Use of the California Mastitis Test for the Detection Of Bovine Mastitis. PMID- 17421324 TI - Feed Additives in Farm Animal Rations. PMID- 17421325 TI - The Preparation and Use of Vaccines for the Prevention of Pasteurellosis. PMID- 17421326 TI - Public Relations. PMID- 17421329 TI - The Veterinarian's Part in National Emergencies. PMID- 17421328 TI - Notes on Technique for Postmortem Examination of the Eye. PMID- 17421330 TI - Tranquilizers in Veterinary Medicine. PMID- 17421331 TI - Attempts at Establishing Swine Herds Free From Atrophic Rhinitis and Virus Pneumonia: 1. Review of Initial Work at the Ontario Veterinary College. PMID- 17421332 TI - Isolation of Salmonella Pullorum from the Joints of Broiler Chickens. PMID- 17421334 TI - Some Notes on Psychology in Harness Racing. PMID- 17421333 TI - Radiology VI. PMID- 17421335 TI - Permanent Intramedullary Pinning. PMID- 17421336 TI - Dystocia in bitches. PMID- 17421337 TI - Bamboo Spine. PMID- 17421338 TI - Osteogenic Sarcoma in a Cat. PMID- 17421340 TI - "SEE ALBERTA" TOUR. PMID- 17421339 TI - Pasteurellosis in Penguins. PMID- 17421342 TI - PERSONAL NEWS. PMID- 17421341 TI - NEWS NOTES. PMID- 17421344 TI - A Study of Bovine Enteric Virus (BEV) Infections Among Cattle on Infected Premises. PMID- 17421345 TI - Parasites and Diseases of Bison in Canada: 1 Tuberculosis and Some Other Pathological Conditions in Bison at Wood buffalo And Elk Island National Parks in the Fall and Winter of 1959-60. PMID- 17421347 TI - Regimen for the Sick Patient. PMID- 17421346 TI - Adrenal Cortical Failure in Dogs. PMID- 17421348 TI - Common Diseases of Laboratory Animals in Canada. PMID- 17421349 TI - PERSONAL NEWS. PMID- 17421350 TI - The Medical Officer's Concept of a Director of Food Control and Sanitation. PMID- 17421351 TI - Learning to Read-Reading to Learn. PMID- 17421353 TI - An Unusual Dystocia. PMID- 17421352 TI - Abomasal Ulceration in a Jersey Cow. PMID- 17421355 TI - NEWS NOTES. PMID- 17421354 TI - Myositis Ossificans Progressiva in a Chicken. PMID- 17421357 TI - An Outbreak of Infectious Bovine Rhinotracheitis in Ontario. PMID- 17421358 TI - Organophosphate Poisoning in Cattle with Particular reference To Co-Ral. PMID- 17421360 TI - PERSONAL NEWS. PMID- 17421359 TI - Simultaneous Immunization of Mink Against Virus Enteritis, Distemper and Botulism. PMID- 17421361 TI - Radiology VII. PMID- 17421362 TI - North American Blastomycosis-A Report of 2 Cases in Dogs. PMID- 17421363 TI - Veterinary Medicine in Alberta-Historical Notes. PMID- 17421364 TI - Tuberculosis in Horses. PMID- 17421366 TI - Traitement Chirurgical d'une Fistule Uretro-Perineale Chez le Chien. PMID- 17421365 TI - Demodicidosis in a Flock of Goats. PMID- 17421367 TI - NEWS NOTES. PMID- 17421369 TI - L'Importance Pratique des Oligoelements Nutritifs. PMID- 17421371 TI - Summertime Diseases of Bovine Feet and Their Treatment. PMID- 17421370 TI - Two Cases of Poisoning in Livestock Presenting Difficulties in Diagnosis. PMID- 17421372 TI - COMMENTS ON A CAREER EXPOSITION. PMID- 17421373 TI - A NOTE ON BANFF ACCOMMODATIONS. PMID- 17421374 TI - An Outline of Feline Medicine. PMID- 17421376 TI - Surgical Intervention in Acute Colic in a Horse. PMID- 17421375 TI - Vaccination Program Against Distemper and Hepatitis. PMID- 17421377 TI - Multiple Sacculated Bronchiectasis in a Lynx. PMID- 17421380 TI - NEWS NOTES. PMID- 17421379 TI - Testicular Tumor in a Budgerigar. PMID- 17421381 TI - Veterinarians and Horse Racing. PMID- 17421383 TI - The Canadian Veterinary Journal. PMID- 17421382 TI - Meat Inspection. PMID- 17421384 TI - An Outbreak of Bovine Malignant Catarrh in a Dairy Herd I: I. Clinical and Pathologic Observations. PMID- 17421385 TI - An Outline of Feline Medicine. PMID- 17421386 TI - Obstructions of the Intestinal Tract in Small Animals Due to Foreign Objects. PMID- 17421388 TI - Two Cases of Poisoning in Livestock Presenting Difficulties in Diagnosis. PMID- 17421389 TI - Johne's Disease in a European Red Deer. PMID- 17421390 TI - Extra-Uterine Pregnancy in a Cow. PMID- 17421391 TI - Chlordane Poisoning in a Cat. PMID- 17421392 TI - News Notes. PMID- 17421393 TI - Personal News. PMID- 17421394 TI - An Outbreak of Bovine Malignant Catarrh in a Dairy Herd. Part II: Transmission Experiments. PMID- 17421395 TI - Problems of Escherichia Coli in Animals and Man. PMID- 17421397 TI - Radiology VIII. PMID- 17421396 TI - Early Ideas on Inbreeding and Crossbreeding. PMID- 17421398 TI - Patent Ductus Arteriosus Offset by Fallot's Syndrome. PMID- 17421400 TI - The Itemized Statement. PMID- 17421399 TI - 75e Anniversaire de l'Ecole de Medecine veterinaire de la Province de Quebec. PMID- 17421401 TI - Facons inusitees d'allaitement de poulains orphelins. PMID- 17421402 TI - Surgical Correction of Blocked Parotid Duct. PMID- 17421403 TI - Cutaneous North American Blastomycosis in a Siamese Cat. PMID- 17421404 TI - Periosteal Fibrosarcoma in a Broiler Chicken. PMID- 17421405 TI - NEWS NOTES. PMID- 17421408 TI - A Discussion of the Common Diseases of Sheep. PMID- 17421410 TI - Pony and Light Horse Practice. PMID- 17421409 TI - Arthritis in Cattle. PMID- 17421411 TI - Diagnosis of Unusual Cases of Avian Mycosis. PMID- 17421412 TI - La Mission Du Veterinaire dans l'industrie de la viande. PMID- 17421413 TI - X-ray Equipment Technics. PMID- 17421414 TI - Mesh Implant for Correction of Congenital Canine Midline Hernia. PMID- 17421415 TI - Large Round Worms in Chicken Eggs. PMID- 17421416 TI - News Notes. PMID- 17421417 TI - Personal News. PMID- 17421419 TI - Coliform Mastitis in Cattle. PMID- 17421421 TI - A Note on Animal Behaviour as a Factor in Parasitism. PMID- 17421420 TI - Histoplasmosis in the Dog: A Report of Four Ontario Cases. PMID- 17421422 TI - Dilatation of the Omasum and Abomasum. PMID- 17421423 TI - Disease Prevention -Excerpts from an address. PMID- 17421424 TI - Congenital Diaphragmatic Hernia in a Calf. PMID- 17421425 TI - Surgical Repair of Medial Patellar Luxations in Dogs. PMID- 17421426 TI - Eosinophilic Myositis in a Beef Carcase. PMID- 17421427 TI - Vaginal Prolapse in a Bitch. PMID- 17421428 TI - Fracture of a Wing in a Falcon. PMID- 17421429 TI - NEWS NOTES. PMID- 17421430 TI - Clostridium Perfringens Enterotoxaemia in the Ruminant. PMID- 17421431 TI - Client Impressions. PMID- 17421432 TI - Comment Je Concois Nos Relations Exterieures. PMID- 17421433 TI - Streptobacillus Moniliformis Infection in a Dog. PMID- 17421434 TI - A Case of Traumatic Reticulitis (Intraparietal Fistula). PMID- 17421435 TI - Le Gluconate de Calcium dans L'Anasarque du Cheval-Resultats d'Experiences Personnelles. PMID- 17421436 TI - Dilatation and Torsion of the Abomasum in a Calf. PMID- 17421438 TI - NEWS NOTES. PMID- 17421437 TI - Suppurative Pneumonia Brought About by Esophageal Perforation in a Puppy. PMID- 17421439 TI - PERSONAL NEWS. PMID- 17421440 TI - B.C. ANIMAL PATHOLOGY LABORATORY REPORT. PMID- 17421442 TI - A New Year's Message from our President. PMID- 17421443 TI - The Effect of Feed-Lot Rations Containing Low-Level Amounts of Aureomycin on the Incidence of Liver Abscesses in Cattle. PMID- 17421444 TI - Address by the President of the A.V.M.A. PMID- 17421445 TI - Two Reports Of Unusual Parasites Diagnosed in Dogs. PMID- 17421446 TI - SUMMARIES OF PAPERS PRESENTED AT C.V.M.A. CONVENTION: THIRTEENTH ANNUAL MEETING. PMID- 17421447 TI - REPORT OF THE EDITORIAL BOARD. PMID- 17421448 TI - The Adrenocortical Family of Hormones and Steroid Therapy. PMID- 17421449 TI - NEWS NOTES. PMID- 17421450 TI - PERSONAL NEWS. PMID- 17421452 TI - INDISCRIMINATE SALE OF ANTIBIOTICS. PMID- 17421451 TI - The Veterinarian in Laboratory Animal Medicine. PMID- 17421453 TI - A typical Interstitial Pneumonia of Cattle. PMID- 17421454 TI - The Application of Specific Pathogen-Free Animals to Research and Production. PMID- 17421455 TI - APOLOGIES! PMID- 17421456 TI - The Advance of Veterinary Science in Great Britain. PMID- 17421457 TI - Adeno-Carcinoma in A Bitch. PMID- 17421458 TI - Observations on the Anthelmintic Effects of Methyridine in Dogs. PMID- 17421459 TI - NEWS NOTES. PMID- 17421461 TI - Nephritis in the Dog. PMID- 17421460 TI - Cultural and Serological Evidence of Leptospirosis in Deer in Ontario. PMID- 17421462 TI - Anomaly of Craniopharyngeal Duct and Hypophysis. PMID- 17421463 TI - Enzymatic Cause of Viscid Ruminal Contents-Its Relation to Bloat. PMID- 17421464 TI - Bovine Mastitis and a Program for Its Control in California. PMID- 17421466 TI - Lead Poisoning in Mink. PMID- 17421465 TI - Ringworm: Its Treatment and Public Health Aspects. PMID- 17421467 TI - THE ONTARIO VETERINARY COLLEGE AT CONFEDERATION. PMID- 17421469 TI - PERSONAL NEWS. PMID- 17421468 TI - NEWS NOTES. PMID- 17421470 TI - The Use of Fluorescent Tracers for Tagging the Flesh of Diseased and Fallen Animals. PMID- 17421471 TI - Bovine Papular Stomatitis. PMID- 17421472 TI - An Electron Microscopic Study Of Leptospira Pomona. PMID- 17421473 TI - Practical Veterinary Hematology. PMID- 17421475 TI - Le Role Social de la Femme du Veterinaire. PMID- 17421474 TI - Suspected Actinomycosis Of Turkeys. PMID- 17421477 TI - NEWS NOTES. PMID- 17421476 TI - Candidia Mastitis in Cows. PMID- 17421478 TI - PERSONAL NEWS. PMID- 17421479 TI - A Technique for Freezing Goat Semen and Results of a Small Breeding Trial. PMID- 17421480 TI - Epizootiology in Disease Investigations. PMID- 17421481 TI - A New Approach to Edematous Conditions of Cattle. PMID- 17421482 TI - Dr. Sanderson's Infallible Oils. PMID- 17421483 TI - NEWS NOTES. PMID- 17421484 TI - PERSONAL NEWS. PMID- 17421485 TI - Briefs. PMID- 17421486 TI - A Method for the Control of Staphylococcal and Streptococcal Mastitis in Ontario. PMID- 17421487 TI - Skin Diseases in Small Animals. PMID- 17421488 TI - Veterinary Instruction at Queen's and O.A.C. PMID- 17421489 TI - Repair of Compound Fracture of the Cannon Bone. PMID- 17421491 TI - NEWS NOTES. PMID- 17421490 TI - Tibial Fracture Repair with Two Jonas Splints. PMID- 17421493 TI - PERSONAL NEWS. PMID- 17421492 TI - Briefs. PMID- 17421494 TI - O.V.C. CENTENNIAL. PMID- 17421495 TI - CENTENAIRE DU COLLEGE VETERINAIRE D'ONTARIO. PMID- 17421496 TI - The Ontario Veterinary College is One Hundred Years Old. PMID- 17421497 TI - Compliments to the Ontario Veterinary College. PMID- 17421498 TI - Tribute to Ontario Veterinary College. PMID- 17421499 TI - Greetings from the President of the British Veterinary Association to the Ontario Veterinary College on its Centenary. PMID- 17421500 TI - Greetings from Mark L. Morris, D.V.M., President American Veterinary Medical Association. PMID- 17421502 TI - Message by Hon. Alvin Hamilton, Minister of Agriculture: On the occasion of the Centennial of the Ontario Veterinary College, July 1962. PMID- 17421501 TI - Message from the President, Royal College of Veterinary Surgeons. PMID- 17421503 TI - Message by Wm. A. Stewart, Minister of Agriculture, Province of Ontario: On the occasion of the Centennial of the Ontario Veterinary College, July, 1962. PMID- 17421504 TI - What is a Veterinarian? PMID- 17421505 TI - Long-Term Survival of Frozen Equine Epididymal Spermatozoa. PMID- 17421506 TI - Demand Feeding of Dogs. PMID- 17421508 TI - VETESCOPE '62. PMID- 17421507 TI - NEWS NOTES. PMID- 17421509 TI - Vibrionic Dysentery of Swine in Ontario-Part I : 1. Clinical Aspects and Pathology. PMID- 17421510 TI - Association of Canine Adenovirus (Toronto A 26/61) with an Outbreak of Laryngotracheitis ("Kennel Cough"): A Preliminary Report. PMID- 17421512 TI - Why is the Modern Poultry Farmer a Sitting Duck? PMID- 17421511 TI - Vitamin A Studies in the Dairy Calf. PMID- 17421513 TI - North American Blastomycosis in a Dog. PMID- 17421515 TI - LETTER TO THE EDITOR. PMID- 17421514 TI - Abdominal Hernia in Turkey Poults. PMID- 17421516 TI - NEWS NOTES. PMID- 17421518 TI - Vibrionic Dysentery in Swine in Ontario-Part II: Morphological, Biochemical and Serological Characteristics of Vibrio Coli. PMID- 17421520 TI - The Shipping Fever Complex. PMID- 17421519 TI - A Comparison of Gastric Torsion with Torsion of the Splenic Pedicle. PMID- 17421521 TI - Clinical Observations on the Use of Tylosin in the Treatment of Vibrionic Swine Dysentery. PMID- 17421522 TI - Western Veterinary College-History and Status 1962. PMID- 17421523 TI - Acute Ancylostomiasis in a litter of Boxer Puppies. PMID- 17421524 TI - Color Changes in the Semi-Tendinosus and the Biceps Femoralis Muscle of Two Hereford Steers. PMID- 17421525 TI - Conservative Treatment for Hydrops in the Cow. PMID- 17421526 TI - Rupture of the Vagina in a Mare During Coitus. PMID- 17421527 TI - NEWS NOTES. PMID- 17421529 TI - Attempts at Establishing Swine Herds Free from Atrophic Rhinitis and Virus Pneumonia: II. Repopulation with Specific Pathogen-Free Pigs. PMID- 17421530 TI - A Note on the Value of Routine Fecal Examination. PMID- 17421531 TI - Polyurethane Polymer-It's Use in Canine Orthopaedics. PMID- 17421532 TI - Geriatrics. PMID- 17421533 TI - A Preliminary Survey on the Incidence of Q Fever Antibodies in the Sera of Manitoba Cattle. PMID- 17421534 TI - Brazilian Groundnut Toxicosis in Canadian Broiler Chickens. PMID- 17421535 TI - A Practising Veterinarian Views the Problems of Veterinary Education. PMID- 17421536 TI - Mycotic Pneumonia in a Pony. PMID- 17421537 TI - Atypical Interstitial Pneumonia of Cattle. PMID- 17421538 TI - LETTERS TO THE EDITOR. PMID- 17421540 TI - NEWS NOTES. PMID- 17421539 TI - FEDERAL MEAT INSPECTION. PMID- 17421541 TI - PERSONAL NEWS. PMID- 17421542 TI - History is the Map of Time. PMID- 17421543 TI - Furaltadone (Valsyn Gel ) Dans Le Traitement des Mammites Bovines. PMID- 17421544 TI - Skin Pigmentation in Buffalo Calves. PMID- 17421545 TI - A Pox-Like Infection of Canaries: Report of an Outbreak. PMID- 17421546 TI - A Simplified Procedure for Intestinal Anastomosis in the Rat. PMID- 17421547 TI - Bovine Tuberculosis in a White-Tailed Deer (Odocoileus virginianus). PMID- 17421549 TI - NEWS NOTES. PMID- 17421548 TI - Metorchis Conjunctus Liver Fluke Infestation in a Cat. PMID- 17421550 TI - PERSONAL NEWS. PMID- 17421552 TI - Furaltadone (Valsyn ) in the Treatment of Bovine Mastitis. PMID- 17421553 TI - Urinary Diversion in Mongrel Dogs: Using Isolated Loops of Ileum for Replacing the Ureter and/or Bladder. PMID- 17421554 TI - Conservation a Longue Echeance du Sperme Epididyme Equin Congele. PMID- 17421555 TI - What is a Veterinarian? Part II. PMID- 17421556 TI - Perforating Ulcers of the Small Intestine-Lymphosarcoma and Fibrosarcoma Complications. PMID- 17421557 TI - Suspected Traumatic Reticulitis in a Cow. PMID- 17421559 TI - PERSONAL NEWS. PMID- 17421558 TI - NEWS NOTES. PMID- 17421561 TI - A New Year's Message from our President. PMID- 17421563 TI - Group Practice in Britain. PMID- 17421562 TI - Thoughts on the Alliance of Veterinary and Human Medicine. PMID- 17421564 TI - Repair of Ruptured Lateral Patellar Ligament. PMID- 17421565 TI - Apparent Blindness in Sows. PMID- 17421566 TI - Primary Hypothyroidism. PMID- 17421567 TI - Fracture of the Cervical Vertebrae in a Heifer. PMID- 17421568 TI - Brucella Eradication in Northern Ireland. PMID- 17421569 TI - NEWS NOTES. PMID- 17421572 TI - PERSONAL NEWS. PMID- 17421570 TI - VETESCOPE '62. PMID- 17421573 TI - Bovine "Enterotoxemia" I. Clostridium Perfringens Types Isolated from Animal Sources in Alberta and Saskatchewan. PMID- 17421574 TI - The Relationship Between Veterinarians and the Feed Industry. PMID- 17421575 TI - A Survey of Salmonella Contamination in Animal Feeds and Feed Constituents. PMID- 17421577 TI - The Operation of a Large Animal Clinic. PMID- 17421576 TI - Swine Pneumonia. PMID- 17421578 TI - Urea Poisoning in Cows at Pasture. PMID- 17421579 TI - Cecalcolic Intussusception in Calf. PMID- 17421580 TI - Typhlitis in Pheasants due to Heterakis Gallinae. PMID- 17421582 TI - NEWS NOTES. PMID- 17421581 TI - Botulism in Ducklings. PMID- 17421584 TI - Immunological Differentiation of Type B and E Strains of Pasteurella Multocida. PMID- 17421585 TI - A Program for Distemper Immunization of Dogs. PMID- 17421586 TI - A Note on Salmonellosis in Adult Cattle Caused by Contaminated Bone Meal. PMID- 17421587 TI - A Bovine Bronchial Fibrinous Cast. PMID- 17421589 TI - NEWS NOTES. PMID- 17421588 TI - Inherited Congenital Hypotrichosis. PMID- 17421590 TI - Disease Reporting in Alberta : Public Relations Committe, Alberta Veterinary Medical Association. PMID- 17421591 TI - Aspergillosis in a Turkey. PMID- 17421592 TI - A Hepatitis-Nephritis Disease of Nutria (Myocastor Coypus) of Probable Viral Etiology. PMID- 17421593 TI - The Use of Volatile Anaesthetic Agents in Horses and Farm Animals. PMID- 17421595 TI - NEWS NOTES. PMID- 17421594 TI - Veterinary Partnership. PMID- 17421596 TI - PERSONAL NEWS. PMID- 17421597 TI - Observations on Animal Behaviour. PMID- 17421598 TI - Control of Gastro-Intestinal Parasites in Monkeys. PMID- 17421599 TI - Saving Money by Estate Planning. PMID- 17421600 TI - Spinal Fracture Plating. PMID- 17421601 TI - The Care of Laboratory Animals. PMID- 17421603 TI - NEWS NOTES. PMID- 17421602 TI - Why Pets? PMID- 17421604 TI - The Use of Adrenal Corticosteroids in Dairy Cattle. PMID- 17421605 TI - Stifle Joint Surgery in the Dog. PMID- 17421606 TI - A Report on the Efficacy of Emtryl on the Prevention and Therapy of Enterohepatitis in Turkeys. PMID- 17421607 TI - Phenothiazine Poisoning in Ponies. PMID- 17421608 TI - A Case of Obstructive Uremia with an Unusual Cause. PMID- 17421609 TI - Thiabenzadole Used Clinically for the Control of Internal Parasites of Sheep. PMID- 17421610 TI - THE UNIVERSITY OF SASKATCHEWAN. PMID- 17421611 TI - PERSONAL NEWS. PMID- 17421613 TI - NEWS NOTES. PMID- 17421615 TI - Greetings to C.V.M.A. PMID- 17421614 TI - THE VETERINARIAN IN SCIENCE. PMID- 17421616 TI - A Discussion of Recent Developments Relating to Pasteurella Hemolytica with Special Reference to Strains Pathogenic for Cattle. PMID- 17421617 TI - Association of Myxovirus Para Influenzae 3 (RE 55) with Upper Respiratory Infection of Horses. PMID- 17421619 TI - NEWS NOTES. PMID- 17421618 TI - Herd Health Practice. PMID- 17421620 TI - LETTERS TO THE EDITOR. PMID- 17421621 TI - LETTERS TO THE EDITOR. PMID- 17421622 TI - A New Look at Swine Feeding Standards. PMID- 17421623 TI - Yellow Tags, Green Paint, and Forms in Triplicate. PMID- 17421624 TI - Collector Tube for Urine and Adapter Therefor. PMID- 17421625 TI - An Outbreak of Diarrhea in a Swine Herd. PMID- 17421626 TI - The Efficacy of Thiabendazole in the Control of Gapeworm Infection in Pheasants. PMID- 17421628 TI - Counterclockwise Abomasal Torsion. PMID- 17421627 TI - Pituitary Abscessation, and a "Farcy-Like" Condition in Cattle Due to Cornebacterium Pyogenes. PMID- 17421629 TI - WESTERN ONTARIO VETERINARY MEDICAL ASSOCIATION. PMID- 17421630 TI - NEWS NOTES. PMID- 17421631 TI - Genetic Aspects of Disease Resistance. PMID- 17421632 TI - Antibiotic Reactions in Cattle. PMID- 17421633 TI - Management of Udder Edema in Cattle. PMID- 17421634 TI - The Collection and Freezing of Semen from an Impotent Bull. PMID- 17421636 TI - LETTER TO THE EDITOR. PMID- 17421635 TI - Posterior Paralysis in a Steer Treated with a Systemic Insecticide for Warble Fly Control. PMID- 17421638 TI - NEWS NOTES. PMID- 17421639 TI - Toxoplasma Microti (The M-Organism) in the Muskrat (Ondatra Zibethica). PMID- 17421641 TI - An Outbreak of Meningitis in Swine Caused by Haemophilus Species of Bacterium. PMID- 17421640 TI - Dalmeny Disease. An Infection of Cattle Presumed to be Caused by an Unidentified Protozoon. PMID- 17421642 TI - Haemophilus Septicemia in Piglets. PMID- 17421643 TI - NEWS NOTES. PMID- 17421644 TI - Canada's Drug Regulations and the Veterinarian. PMID- 17421645 TI - Anthrax in Canada. PMID- 17421646 TI - Bovine "Enterotoxemia". II. Experimental Reproduction of the Disease. PMID- 17421648 TI - Clinical Experiences with Clostridia. PMID- 17421647 TI - North American Blastomycosis in a Dog. PMID- 17421649 TI - PERSONAL NEWS. PMID- 17421650 TI - VETESCOPE, '63. PMID- 17421652 TI - NEWS NOTES. PMID- 17421653 TI - Presidential Address. PMID- 17421654 TI - Responsibility of a Veterinarian to his Clients. PMID- 17421655 TI - Plants Poisonous to Livestock. PMID- 17421656 TI - Bacteriological and Pathological Studies of Natural and Experimental Swine Abortion Due to Leptospira Pomona. PMID- 17421658 TI - EXCERPTS FROM A TALK BY DEAN W. W. ARMISTEAD. PMID- 17421657 TI - An Outbreak of Salmonellosis in Mature Dairy Cattle. PMID- 17421660 TI - NEWS NOTES. PMID- 17421661 TI - LETTER TO THE EDITOR. PMID- 17421662 TI - A New Year's Message from our President. PMID- 17421663 TI - Donnees sur la Composition en Nitrates et Carotenes des Plantes Herbageres Traitees a l'Herbicide MCPA. PMID- 17421664 TI - Leukemia in a Cat. PMID- 17421665 TI - Editorial. PMID- 17421666 TI - The Use of Percussion and Auscultation as a Diagnostic Aid in Abomasal Displacement of Dairy Cows. PMID- 17421667 TI - Observations on an Outbreak of Clinical Parasitism in Ewes During the Winter Months. PMID- 17421669 TI - NEWS NOTES. PMID- 17421670 TI - Editorial. PMID- 17421671 TI - Recent Developments in the Control of Arthropod Parasites of Cattle. PMID- 17421672 TI - A Clinical and Laboratory Evaluation of a New Sulfonamide. PMID- 17421673 TI - Mesenteric Aneurism in a Horse. PMID- 17421674 TI - Pregnancy in a Hermaphrodite Sow. PMID- 17421675 TI - LETTER TO THE EDITOR. PMID- 17421677 TI - NEWS NOTES. PMID- 17421678 TI - Editorial. PMID- 17421679 TI - Abomasal Abnormalities in Dairy Cattle: A Review of 90 Clinical Cases. PMID- 17421680 TI - Two Unusual Orthopaedic Prostheses. PMID- 17421681 TI - An Attempt to Eliminate Haemonchus Contortus from a Sheep Flock in Ontario. PMID- 17421682 TI - An Unusual Outbreak of Sweet Clover Poisoning in Cattle. PMID- 17421683 TI - LETTERS TO THE EDITOR. PMID- 17421684 TI - NEWS NOTES. PMID- 17421685 TI - Editorial. PMID- 17421687 TI - Propagation of Rabies Virus in Pig Kidney Cell Culture. PMID- 17421686 TI - The Clinical Diagnosis of Rabies in Cattle. PMID- 17421688 TI - Thrombosis of the Spermatic Artery Resembling Torsion of the Spermatic Cord in a Stallion. PMID- 17421689 TI - LETTERS TO THE EDITOR. PMID- 17421691 TI - LETTERS TO THE EDITOR. PMID- 17421690 TI - LETTERS TO THE EDITOR. PMID- 17421692 TI - PERSONAL NEWS. PMID- 17421693 TI - NEWS NOTES. PMID- 17421695 TI - The Growth of Meat Inspection in Canada. PMID- 17421696 TI - Salivary Mucoceles in Dogs. PMID- 17421697 TI - Danger of Residual Strychnine in Syringes. PMID- 17421698 TI - Editorial. PMID- 17421699 TI - Ontario Certified Herd Policy for Swine. PMID- 17421700 TI - PERSONAL NEWS. PMID- 17421701 TI - NEWS NOTES. PMID- 17421703 TI - Editorial. PMID- 17421704 TI - Wildlife and Some Diseases of Domestic Animals in Canada. PMID- 17421705 TI - Development of the Club Practice in New Zealand and Some Thoughts on Its Future. PMID- 17421706 TI - The Role of the Cerebrospinal Fluid Tap in the Neurological Examination of the Dog. PMID- 17421707 TI - Group Small Animal Practice. PMID- 17421708 TI - LETTERS TO THE EDITOR. PMID- 17421709 TI - LETTERS TO THE EDITOR. PMID- 17421710 TI - LETTERS TO THE EDITOR. PMID- 17421713 TI - NEWS NOTES. PMID- 17421711 TI - LETTERS TO THE EDITOR. PMID- 17421714 TI - Editorial. PMID- 17421715 TI - Greetings to C.V.M.A. PMID- 17421716 TI - Colibacillosis of Calves in Ontario: I. A Serological Study of Escherichia Coli Strains. PMID- 17421718 TI - Entero-Typhlitis of Broiler Chicks. PMID- 17421717 TI - Modification du Comportement et de la Taille du Vison par la Reserpine et le Methandrostenolone. Etude Clinique. PMID- 17421719 TI - NEWS NOTES. PMID- 17421720 TI - Editorial. PMID- 17421721 TI - Bovine Lymphosarcoma. A Review of Studies on Cattle in the Eastern United States. PMID- 17421722 TI - Fluid and Electrolyte Therapy in Small Animals. PMID- 17421724 TI - LETTERS TO THE EDITOR. PMID- 17421723 TI - LETTERS TO THE EDITOR. PMID- 17421725 TI - LETTERS TO THE EDITOR. PMID- 17421727 TI - Editorial. PMID- 17421726 TI - NEWS NOTES. PMID- 17421728 TI - Use of Thiamylal Sodium in Mature Swine. PMID- 17421729 TI - Colibacillosis of Calves in Ontario. II. The Association of Certain Serotypes of Escherichia Coli with Calf Scours. PMID- 17421730 TI - Proper Care of Wounds in Horses. PMID- 17421731 TI - Problems Related to Vaccination in the Canadian Brucellosis Control Program. PMID- 17421732 TI - The Surgical Treatment of a Persistent Right Aortic Arch and Patent Ductus Arteriosus in a Dog. PMID- 17421733 TI - LETTER TO THE EDITOR. PMID- 17421734 TI - LETTER TO THE EDITOR. PMID- 17421735 TI - NEWS NOTES. PMID- 17421736 TI - Editorial. PMID- 17421737 TI - Studies on Colibacillosis of Calves. I. The Antibody Acquired by Calves as the Result of Vaccination of the Dam. PMID- 17421738 TI - Control of Disease in Dogs in the Canadian North. PMID- 17421739 TI - The Effect of Prophylactic Treatment of Pregnant Beef Cows on the Incidence of Nutritional Muscular Dystrophy. A Field Trial. PMID- 17421741 TI - BURSITE NAVICULAIRE SEPTIQUE DU PIED DES BOVINS. PMID- 17421740 TI - The Use of the Scalp Vein Infusion in Dogs. PMID- 17421742 TI - Giant Kidney Worm in Canine Abdomen. PMID- 17421744 TI - NEWS NOTES. PMID- 17421743 TI - Lead Poisoning in Calves. PMID- 17421745 TI - An Attenuated Rabies Vaccine for Domestic Animals Produced in Tissue Culture. PMID- 17421746 TI - Elaphostrongylosis in Maritime Moose and Deer. PMID- 17421747 TI - Studies on Colibacillosis of Calves: II. A Clinical Evaluation of the Efficiency of Vaccination of the Dam as a Means of Preventing Colibacillosis of the Calf. PMID- 17421748 TI - PERSONAL NEWS. PMID- 17421749 TI - NEWS NOTES. PMID- 17421750 TI - PROMINENT VETERINARY DIES. PMID- 17421752 TI - Presidential Address. PMID- 17421753 TI - Studies on Colibacillosis of Calves. III. The Experimental Reproduction of Colibacillosis. PMID- 17421754 TI - Distemper in Raccoons Characterized by Giant-Cell Pneumonitis. PMID- 17421756 TI - NEWS NOTES. PMID- 17421755 TI - On the Efficacy of Thiabendazole Against Immature Oesophagostomum Columbianum in Sheep. PMID- 17421757 TI - A New Year's Message from our President. PMID- 17421758 TI - LETTER TO THE EDITOR. PMID- 17421759 TI - NEWS NOTES. PMID- 17421763 TI - NEWS NOTES. PMID- 17421761 TI - LETTER TO THE EDITOR. PMID- 17421764 TI - LETTER TO THE EDITOR. PMID- 17421765 TI - Editorial. PMID- 17421767 TI - NEWS NOTES. PMID- 17421766 TI - Herniated Pregnancy in a Dog. PMID- 17421768 TI - NEWS NOTES. PMID- 17421770 TI - NEWS NOTES. PMID- 17421769 TI - Immunization of the Dog. PMID- 17421772 TI - Greetings to C.V.M.A. PMID- 17421773 TI - LETTERS TO THE EDITOR. PMID- 17421775 TI - NEWS NOTES. PMID- 17421776 TI - PERSONAL NEWS. PMID- 17421777 TI - Editorial. PMID- 17421780 TI - NEWS NOTES. PMID- 17421778 TI - NEWS NOTES. PMID- 17421781 TI - NEWS NOTES. PMID- 17421782 TI - LETTERS TO THE EDITOR. PMID- 17421783 TI - LETTERS TO THE EDITOR. PMID- 17421784 TI - LETTERS TO THE EDITOR. PMID- 17421785 TI - NEWS NOTES. PMID- 17421786 TI - PERSONAL NEWS. PMID- 17421787 TI - Presidential Address. PMID- 17421789 TI - NEW PRODUCT. PMID- 17421788 TI - LETTER TO THE EDITOR. PMID- 17421790 TI - A New Year's Message from Our President. PMID- 17421791 TI - Vaccination of Swine with erysipelas vaccine (live culture-modified) by the oral route. PMID- 17421793 TI - Letter to the editor. PMID- 17421792 TI - Successful surgical treatment of bilateral renal calculi in a dog. PMID- 17421794 TI - Letter to the editor. PMID- 17421796 TI - News notes. PMID- 17421795 TI - The hematology of phenothiazine poisoning in horses. PMID- 17421798 TI - Common diseases of fur bearing animals I. Diseases of mink. PMID- 17421799 TI - Helminthiasis in cattle and failure of coumaphos to control it. PMID- 17421800 TI - Coligranuloma (Hjarre's Disease) in Turkeys. PMID- 17421801 TI - Sarcoptic scabies in a dog (with human involvement). PMID- 17421802 TI - Note sur la biopsie du grand epiploon. PMID- 17421803 TI - Letters to the editor. PMID- 17421804 TI - Letters to the editor. PMID- 17421805 TI - News notes. PMID- 17421807 TI - Trends in veterinary education. PMID- 17421808 TI - The veterinarian in provincial veterinary services. PMID- 17421809 TI - Veterinary medical education for the next decade. PMID- 17421810 TI - The Western canadian veterinary college-its philosophy. PMID- 17421811 TI - The veterinarian and animal production. PMID- 17421812 TI - Veterinary education. PMID- 17421813 TI - Letter to the editor. PMID- 17421814 TI - News notes. PMID- 17421815 TI - Letter to the editor. PMID- 17421816 TI - News notes. PMID- 17421818 TI - News notes. PMID- 17421820 TI - News notes. PMID- 17421821 TI - Greetings to C.v.m.a. PMID- 17421822 TI - Occurrence of estrus in an ovariohysterectomized cat. PMID- 17421823 TI - Quonundra. PMID- 17421824 TI - Letters to the editor. PMID- 17421826 TI - News notes. PMID- 17421825 TI - Letters to the editor. PMID- 17421827 TI - Editorial. PMID- 17421828 TI - Quonundra. PMID- 17421829 TI - Letter to the editor. PMID- 17421830 TI - News notes. PMID- 17421832 TI - Quonundra. PMID- 17421833 TI - News notes. PMID- 17421834 TI - Quonundra. PMID- 17421835 TI - News notes. PMID- 17421836 TI - Letter to the editor. PMID- 17421838 TI - Personal news. PMID- 17421837 TI - Quonundra. PMID- 17421839 TI - News notes. PMID- 17421840 TI - New product. PMID- 17421842 TI - Quonundra. PMID- 17421841 TI - The fate of donor cattle cells following transfusion. PMID- 17421843 TI - News notes. PMID- 17421844 TI - A New Year's Message from our President. PMID- 17421845 TI - A report of a natural outbreak of botulism in pheasants. PMID- 17421846 TI - Quonundra. PMID- 17421847 TI - News notes. PMID- 17421849 TI - News notes. PMID- 17421848 TI - Quonundra. PMID- 17421851 TI - Personal news. PMID- 17421850 TI - Television is kicking your image around. PMID- 17421852 TI - Quonundra. PMID- 17421853 TI - News notes. PMID- 17421854 TI - New product. PMID- 17421855 TI - Quonundra. PMID- 17421856 TI - News notes. PMID- 17421857 TI - New product. PMID- 17421858 TI - Letter to the editor. PMID- 17421859 TI - Letter to o.v.C. Class presidents. PMID- 17421860 TI - Quonundra. PMID- 17421861 TI - News notes. PMID- 17421862 TI - Quonundra. PMID- 17421864 TI - Greetings to the C.v.m.a. PMID- 17421863 TI - News notes. PMID- 17421865 TI - New developments in Swine feeding. PMID- 17421866 TI - Antibiotic resistance trends of animal pathogens in ontario. PMID- 17421868 TI - Letters to the editor. PMID- 17421867 TI - Thiabendazole as an Effective Agent Against Giardia SPP in Chinchillas. PMID- 17421870 TI - Quonundra. PMID- 17421869 TI - Letters to the editor. PMID- 17421871 TI - News notes. PMID- 17421872 TI - Radiation therapy in veterinary medicine. PMID- 17421873 TI - The future of large animal practice in alberta. PMID- 17421874 TI - Electroencephalography with barbiturate anesthesia in the dog. PMID- 17421876 TI - Rumenitis in cattle. PMID- 17421875 TI - Atrophic rhinitis-epidemiology and effect of the disease on maturity time of market pigs. PMID- 17421877 TI - Quonundra. PMID- 17421878 TI - News notes. PMID- 17421879 TI - Quonundra. PMID- 17421880 TI - News notes. PMID- 17421881 TI - Quonundra. PMID- 17421882 TI - News notes. PMID- 17421883 TI - Personal news. PMID- 17421884 TI - Letters to the editor. PMID- 17421885 TI - Letters to the editor. PMID- 17421887 TI - News notes. PMID- 17421886 TI - News notes. PMID- 17421889 TI - A New Year's Message from our President. PMID- 17421890 TI - Dermatophilus Dermatitis (Streptotrichosis) in Ontario. II. Laboratory Findings. PMID- 17421891 TI - A new influenza virus infection in turkeys. I. Isolation and characterization of virus 6213. PMID- 17421893 TI - News notes. PMID- 17421892 TI - Suggested changes in veterinary education. PMID- 17421895 TI - News notes. PMID- 17421896 TI - News notes. PMID- 17421897 TI - Letter to the editor. PMID- 17421898 TI - News notes. PMID- 17421899 TI - C.v.m.a. Milestone. PMID- 17421900 TI - News notes. PMID- 17421903 TI - New product. PMID- 17421902 TI - News notes. PMID- 17421905 TI - Quonundra. PMID- 17421904 TI - Greetings to the C. V. M. A. PMID- 17421906 TI - News notes. PMID- 17421907 TI - Letter to the editor. PMID- 17421908 TI - News notes. PMID- 17421910 TI - News notes. PMID- 17421909 TI - News notes. PMID- 17421911 TI - Letters to the editor. PMID- 17421912 TI - Letters to the editor. PMID- 17421913 TI - News notes. PMID- 17421914 TI - Letter to the editor. PMID- 17421916 TI - News notes. PMID- 17421915 TI - Non-surgical extraction of teeth in small primates. PMID- 17421917 TI - A New Year's Message from our President. PMID- 17421918 TI - The nephrosis-nephritis syndrome in chickens caused by a holte strain of infectious bronchitis virus. PMID- 17421919 TI - News notes. PMID- 17421920 TI - Editorial. PMID- 17421921 TI - News notes. PMID- 17421922 TI - Watson and gallivan discovery commemorated by plaque. PMID- 17421923 TI - Food for thought. PMID- 17421925 TI - Collecting bargaining for professional veterinarians. PMID- 17421924 TI - Cvma hospital inspection. PMID- 17421926 TI - News notes. PMID- 17421927 TI - News notes. PMID- 17421928 TI - News notes. PMID- 17421930 TI - News notes. PMID- 17421929 TI - Food for thought. PMID- 17421932 TI - Letter to the editor. PMID- 17421931 TI - News notes. PMID- 17421934 TI - News notes. PMID- 17421933 TI - The american horse council. PMID- 17421935 TI - News notes. PMID- 17421936 TI - News notes. PMID- 17421937 TI - News notes. PMID- 17421938 TI - Animal health technicians - a practice asset. PMID- 17421939 TI - News notes. PMID- 17421940 TI - Message du president. PMID- 17421941 TI - News notes. PMID- 17421943 TI - News notes. PMID- 17421942 TI - Guest editorial. PMID- 17421944 TI - News notes. PMID- 17421945 TI - Letter to the editor. PMID- 17421946 TI - Message du president. PMID- 17421947 TI - Streptothricosis in Saskatchewan. PMID- 17421949 TI - Letters to the editor. PMID- 17421948 TI - Infectivity of d. Viviparus (moose strain) to calves. PMID- 17421950 TI - Anesthetic abnormality. PMID- 17421951 TI - Rapport de l'ACV au Congres de l'agriculture canadienne. PMID- 17421952 TI - L'Apport du medecin veterinaire dans une industrie colossale. PMID- 17421953 TI - A letter to the association. PMID- 17421954 TI - Urinary calculi in puppies. PMID- 17421955 TI - Hepato-nephrose puerperale bovine. PMID- 17421956 TI - Letters to the editor. PMID- 17421957 TI - Message du president. PMID- 17421958 TI - Stringhalt in a calf. PMID- 17421959 TI - Cheyletiella species. PMID- 17421960 TI - Aslap welcomes canadian members. PMID- 17421961 TI - Clinical note: pyometra in a nine-year-old labrador retriever. PMID- 17421962 TI - Letters to the editor. PMID- 17421963 TI - Letters to the editor. PMID- 17421964 TI - Letter to the editor. PMID- 17421965 TI - Letter to the editor. PMID- 17421966 TI - From Finland. PMID- 17421967 TI - The instant animal florence nightingales. PMID- 17421968 TI - Letters to the editor. PMID- 17421969 TI - Message du president. PMID- 17421970 TI - Letters to the editor. PMID- 17421971 TI - Letters to the editor. PMID- 17421972 TI - Letters to the editor. PMID- 17421973 TI - Letters to the editor. PMID- 17421974 TI - Letters to the editor. PMID- 17421975 TI - Orthopedic problem. PMID- 17421976 TI - Letter to the editor. PMID- 17421978 TI - The livestock industry and the veterinarian. PMID- 17421977 TI - Message du president. PMID- 17421979 TI - Silent sapiens. PMID- 17421980 TI - Mycoplasma genital tract infections in cattle. PMID- 17421981 TI - Letters to the editor. PMID- 17421982 TI - Letters to the editor. PMID- 17421983 TI - La rhino-tracheite infectieuse au Quebec: enquete-serologique chez les bovins laitiers. PMID- 17421984 TI - Letters to the editor. PMID- 17421985 TI - Letters to the editor. PMID- 17421986 TI - Letters to the editor. PMID- 17421987 TI - Letters to the editor. PMID- 17421988 TI - Orthopedic problem. PMID- 17421989 TI - Letters to the editor. PMID- 17421990 TI - Letters to the editor. PMID- 17421991 TI - Letters to the editor. PMID- 17421993 TI - Message du president. PMID- 17421992 TI - Letters to the editor. PMID- 17421995 TI - American association of equine practitioners continued growth and progress in 1974. PMID- 17421994 TI - Stomach injection pump for large animals. PMID- 17421996 TI - OVC Expansion Grant. PMID- 17421998 TI - Why not hire a female veterinarian? PMID- 17421997 TI - International society of biometeorology. PMID- 17421999 TI - Yukon needs veterinary services. PMID- 17422000 TI - President's Message. PMID- 17422001 TI - Future of the veterinary profession in Canada. PMID- 17422003 TI - Share your Experience. PMID- 17422002 TI - Nursing care in veterinary medicine. PMID- 17422004 TI - A personal tribute. PMID- 17422005 TI - Expansion of veterinary manpower in Canada. PMID- 17422006 TI - Veterinary manpower in Canada. PMID- 17422007 TI - Implication of expanded veterinary education in Canada. PMID- 17422008 TI - Veterinary education in Canada. PMID- 17422009 TI - Appeal to readers. PMID- 17422011 TI - Treatment of osteomyelitis in a horse. PMID- 17422010 TI - Dalmeny disease-another outbreak-probably sarcocystosis. PMID- 17422012 TI - Treatment of osteomyelitis in a horse: a reply. PMID- 17422014 TI - Air transportation of animals. PMID- 17422013 TI - Manpower shortage? PMID- 17422016 TI - President's Message. PMID- 17422015 TI - Registration of dogs. PMID- 17422018 TI - Disease problems related to type of food. PMID- 17422019 TI - Canadian association of veterinary anatomists: abstracts. PMID- 17422020 TI - Veterinary manpower. PMID- 17422022 TI - Keeping informed. PMID- 17422021 TI - Women in the veterinary profession. PMID- 17422023 TI - Debater's Views Questioned. PMID- 17422024 TI - Veterinary manpower requirements. PMID- 17422025 TI - Views defended by debater. PMID- 17422026 TI - Poultry diseases diagnosed in canadian laboratories for the year 1975. PMID- 17422027 TI - Medial aspects of the canine tarsus. PMID- 17422028 TI - How to handle complaints. PMID- 17422029 TI - Soap poisoning in a dog. PMID- 17422031 TI - Comment on manpower. PMID- 17422030 TI - Renal oxalosis in the cat. PMID- 17422032 TI - Clinical coccidiosis. PMID- 17422034 TI - Poultry diseases diagnosed in canadian laboratories for the year 1976. PMID- 17422033 TI - The canadian veterinary medical association - democracy or oligarchy? PMID- 17422035 TI - The abuse of wildlife immobilizing drugs. PMID- 17422036 TI - Brucellosis and wildlife. PMID- 17422037 TI - Tested tips for interviewing job applicants. PMID- 17422038 TI - The canadian veterinary medical association - democracy or oligarchy: a reply from the president. PMID- 17422040 TI - The canadian veterinary medical association - democracy or oligarchy: a member comments. PMID- 17422039 TI - Drug resistance in avian coccidia. PMID- 17422041 TI - The canadian veterinary medical association - democracy or oligarchy: a reply. PMID- 17422042 TI - Use of fungassay medium in the diagnosis of ringworm. PMID- 17422043 TI - Ovine Abortion due to Chlamydia in Ontario. PMID- 17422044 TI - Reponse a la lettre a l'editeur du Dr Lord. PMID- 17422045 TI - Letters to the editor to contain practice information. PMID- 17422046 TI - Cutaneous melanoma in hampshire Swine. PMID- 17422047 TI - An unusual case of acute abdomen produced by torsion of a retained testicle in a boxer. PMID- 17422049 TI - Handyman's Suction Apparatus. PMID- 17422048 TI - Drug purchasing cooperative. PMID- 17422050 TI - Progesterone therapy for cats. PMID- 17422051 TI - Cat and small dog insulin diluent. PMID- 17422052 TI - Negative image projected. PMID- 17422053 TI - Canadian veterinarians and international development. PMID- 17422054 TI - Understanding animal behaviour. PMID- 17422055 TI - Author of "the dog crisis" lists her credentials. PMID- 17422056 TI - Reviewer of "the dog crisis" replies. PMID- 17422057 TI - "For not Looking than not Knowing". PMID- 17422059 TI - Heartworm in dogs in Canada in 1978. PMID- 17422058 TI - Egg binding in a Western painted turtle. PMID- 17422060 TI - Cat and small dog insulin diluent - a correction. PMID- 17422062 TI - The role of the professional association. PMID- 17422061 TI - A plea for broader advertising of positions in international veterinary medicine. PMID- 17422063 TI - The laws of the land. PMID- 17422064 TI - Practical orthopedic power tools and gas sterilization. PMID- 17422065 TI - Embryo transfer: a discussion on its potential for infectious disease control based on a review of studies on infection of gametes and early embryos by various agents. AB - Studies on laboratory animals have shown that viruses vary as to whether or not they are transmissible by the gametes or are capable of passing through the zona pellucida and infecting the embryo.Methods of studying early embryos for the presence of infectious agents include electron microscopy, immunocytochemistry and cell cultivation.Determination that early bovine embryos do not become infected by certain agents might allow for easing of restrictions in the current import and export regulations for cattle embryos.Embryo transfer could be used as a means of controlling or eliminating disease in a herd or flock if the causal agent does not infect the early embryo via the gametes or by penetrating the zona pellucida. PMID- 17422066 TI - Small animal medicine - advertising - ethics. PMID- 17422067 TI - Campylobacter enteritis in dogs and cats. PMID- 17422068 TI - Canine parvoviral enteritis: a disease entity. PMID- 17422069 TI - Hormone Treatment of Non-productive Bitches. PMID- 17422070 TI - Pansteatitis in Cats Fed Fish-based Commercial Foods. PMID- 17422071 TI - Canine artificial insemination. PMID- 17422072 TI - Fertilizer Poisoning - Is it a Problem? PMID- 17422073 TI - Pet foods in Canada. PMID- 17422074 TI - Practical vacuum system for rapid blood collection. PMID- 17422075 TI - Protestation contre le mauvais francais dans les pages jaunes de la Revue veterinaire canadienne. PMID- 17422076 TI - Coccidiosis in chickens recorded in alberta between 1973 and 1977. PMID- 17422077 TI - Recommendations for slide presentations at meetings. PMID- 17422079 TI - British columbia views on the canadian veterinary journal. PMID- 17422078 TI - Tribute to j.R. Kinney. PMID- 17422080 TI - Isolation of a Hamster-Lethal Strain of Leptospira hardjo. PMID- 17422081 TI - Response to "comments on update on pasteurellosis in young cattle". PMID- 17422082 TI - A plea against ear cropping. PMID- 17422083 TI - An international association of veterinary librarians to serve the veterinary profession. PMID- 17422084 TI - Fluorescent Antibody Test for Rabies Screening is Questioned. PMID- 17422085 TI - Discovery of a Coagulase from Erysipelothrix rhusiopathiae. PMID- 17422086 TI - Correct Syntax is Part of a Good Scientific Paper. PMID- 17422087 TI - Current status of various parvovirus vaccines. PMID- 17422088 TI - Rabies prophylaxis in humans - latest recommendations. PMID- 17422089 TI - Diagnostic and therapeutic tips on cardiology. PMID- 17422090 TI - The present status of euthanasia by nonanesthetic gases. PMID- 17422091 TI - Mastitis indicator. PMID- 17422093 TI - A biography of john cochrane smith 1885 - 1917. PMID- 17422092 TI - D.L.T. Smith: a short biography. PMID- 17422094 TI - Cost effectiveness of methods for treatment of grain overload in cattle. PMID- 17422096 TI - Vaccination guidelines for dogs and cats. PMID- 17422095 TI - Letters to the editor. PMID- 17422097 TI - Intermediary Host for Fasciola hepatica Identified in Quebec. PMID- 17422098 TI - A Simple and Inexpensive Medium for the Cultivation of Erysipelothrix rhusiopathiae. PMID- 17422099 TI - On the multifactorial etiology of Swine pneumonia. PMID- 17422100 TI - Sow culling survey in alberta. PMID- 17422101 TI - Early spay-neutering of dogs and cats. PMID- 17422103 TI - Fall abortions in sows. PMID- 17422102 TI - Bovine Leukosis is Now Only a Sporadic Problem in Ireland. PMID- 17422104 TI - Lyophilized gamma globulin from blood of healthy slaughtered Swine: an effective agent for the prevention and treatment of pig scours. PMID- 17422105 TI - Laparohysterectomy as a treatment for uterine rupture in the cow. PMID- 17422106 TI - More on economics in bovine practice. PMID- 17422107 TI - Systemic d-phenylalanine and d-leucine for effective treatment of pain in the horse. AB - This study showed that subcutaneous injection of a solution of D-amino acids produced effective analgesia in horses. It is postulated that systemic D phenylalanine and D-leucine may become one of the safe, effective and nonaddictive drugs for acute and chronic pain treatment. These D-amino acids cause analgesia by presumably preserving brain endorphins. They may bind reversibly to enkephalinases and prevent enzymatic degradation of enkephalins. PMID- 17422108 TI - Cubitus-curvus chez une chienne. AB - CUBITUS-CURVUS IN A BITCH: This paper presents a case of cubitus-curvus seen in a five month old female crossbreed dog. There is a description of the clinical and radiological aspects of the case and the corrective surgery is also well described. Special attention was given to the normal anatomy and physiology of the growth plates and to the traumas that can affect these structures. Finally, there is an explanation of the pathogeny of the cubitus-curvus. In our case, the evolution was favorable and the healing of the condition was complete. PMID- 17422109 TI - Feline Cutaneous Phaeohyphomycosis Caused by Phialophora verrucosa. AB - A cutaneous granuloma which developed on the muzzle of a nine year old cat was shown by histopathology to have been caused by a dematiaceous fungus which was cultured and identified as Phialophora verrucosa. PMID- 17422110 TI - An Evaluation of a Teat Dip with Dodecyl Benzene Sulfonic Acid in Preventing Bovine Mammary Gland Infection from Experimental Exposure to Streptococcus agalactiae and Staphylococcus aureus. AB - The effectiveness of a teat dip with dodecyl benzene sulfonic acid (1.94%) for the prevention of intramammary infections was determined in cows experimentally challenged with Streptococcus agalactiae and Staphylococcus aureus. The infection rates with Streptococcus agalactiae and Staphylococcus aureus were 62.5% and 75% in undipped quarters, 12.5% and 21.5% in dipped quarters with a reduction rate of 80% and 71% respectively. The significance of some findings in relation to mastitis control are discussed. PMID- 17422111 TI - Johne's Disease (Paratuberculosis) in Goats: A Report of Eight Cases in Quebec. AB - Johne's disease (paratuberculosis) is reported in eight goats. Clinical signs were mainly those of a chronic wasting disease which lead to emaciation, severe weakness, prostration and death. Soft pasty stool and/or profuse diarrhea were observed only after several weeks of illness, in the terminal stages of the disease, in six of the eight goats. Gross lesions were characterized by emaciation and mesenteric lymphadenopathy with more or less extensive areas of caseous necrosis and calcification. Gross lesions in the intestinal tract were limited to a mild thickening and corrugation of the mucosa of the distal small intestine, cecum and proximal parts of the colon in three of the affected animals. Caseous necrosis of Peyer's patches and ulceration were observed in two goats when small tuberculoid nodules were present in the liver of another one. Histologically, the intestinal lesions were characterized by accumulations of large foamy or vacuolated macrophages in the intestinal mucosa. Lesions were usually mild and multifocal in the proximal and middle parts of the small intestine, more diffuse and severe in the distal jejunum, ileum and proximal parts of the colon. Several of the macrophages infiltrated in the intestinal mucosa and mesenteric lymph nodes were packed with acid fast bacilli. Observations made on these animals stress the importance of a careful histological examination of the intestines and mesenteric lymph nodes of adult goats submitted to necropsy because of a chronic wasting disease. PMID- 17422112 TI - Aortic-iliac thrombosis in a horse. AB - A horse with a history of chronic lameness was presented with signs of abdominal pain. A diagnosis of intestinal obstruction was initially made and treatment was ineffective. Further examination revealed an aortic obstruction in the area of the bifurcation of the iliac vessels. Postmortem results supported the clinical findings. The clinical signs of aortic-iliac thrombosis are consistent with the lameness pattern and abdominal distress. PMID- 17422113 TI - Johne's Disease (Paratuberculosis) in a Goat. AB - Johne's disease (paratuberculosis) was diagnosed as the cause of chronic weight loss and intermittent diarrhea in a five year old Saanen doe. Confirmatory necropsy findings included granulomatous enteritis, lymphadenitis, lymphangitis and the demonstration of abundant acid fast organisms within macrophages in impression smears of intestinal mucosa. Some of the difficulties encountered in diagnosing and controlling Johne's disease are discussed with emphasis given to the disease in small ruminants. PMID- 17422114 TI - Exercise in diagnostic radiology. PMID- 17422115 TI - Copper toxicosis in lambs fed milk replacer. AB - Thirty-four hysterectomy derived, crossbred lambs were fed a commercial, lamb milk replacer, containing added copper from birth. Twenty-five lambs died, four were killed and five survived. At necropsy, generalized icterus, enlarged kidneys and enlarged or small livers were found.CLINICAL PATHOLOGY FINDINGS INCLUDED: responsive hemolytic anemia and occasional spherocytosis, hemoglobinemia, hemoglobinuria and urinary casts, leukocytosis with neutrophilia, a liver copper content range of 38-584 ppm; a kidney copper content 6-86 ppm, and serum aspartate amino-transferase level range of 150-302 I.U./L. Histopathologically there was either periacinar hepatic necrosis or widespread portal fibrosis and nephrosis.The role of age and stress in the development of toxicosis is discussed. PMID- 17422116 TI - Enzootic ataxia (swayback) in goats. PMID- 17422117 TI - Exercise in diagnostic radiology. PMID- 17422119 TI - Giardiasis in two calves. PMID- 17422118 TI - Research and the real world. PMID- 17422120 TI - Un epithelioma pulmonaire anaplasique chez une chienne. AB - A PULMONARY ANAPLASTIC EPITHELIOMA IN A BITCH: An anaplastic epithelioma of the lungs was diagnosed in a 26 month old female Airedale. The authors describe signs of bronchopneumonia and lameness involving the right fore and hind limbs. The lameness occurred following metastases to the long bones. The clinical signs, the radiological appearance, the postmortem lesions encountered with that neoplasm and differential diagnosis are discussed. PMID- 17422121 TI - Enzootic nasal adenocarcinoma of sheep in Canada. AB - A survey of veterinary diagnostic laboratories revealed that intranasal tumors occur in sheep in most provinces of Canada. Tumors were diagnosed in 44 sheep of several breeds including Polled Dorset, Suffolk, Cheviot, Rambouillet and various crossbreeds. Twenty-seven percent of tumors occurred in sheep that were less than two years old. Most tumors were sporadic but 33% of cases occurred in six related flocks, indicating that this disease can be an enzootic problem. The clinical signs were persistent serous, mucous or mucopurulent nasal discharge and stridor. Affected sheep progressively developed anorexia, dyspnea and mouth breathing and most died from effects of asphyxia and inanition within 90 days of the onset of clinical signs. Tumors originated unilaterally or occasionally bilaterally in the olfactory mucosa of the ethmoid turbinates. They were expansive and sometimes locally invasive but metastases were not found. Histologically, the tumors were classified as adenomas or, more frequently, adenocarcinomas. The etiology was not established but retrovirus like particles were observed in tumor tissue from one affected sheep. PMID- 17422122 TI - Local veterinary needs poorly perceived by outsiders. PMID- 17422123 TI - Streptococcus suis Type 2 Infection in Swine in Ontario: A Review of Clinical and Pathological Presentations. AB - Over an 18 month period Streptococcus suis type 2 was isolated in pure or mixed culture in 19 disease outbreaks in pigs. Morbidity and case fatality were variable. Clinical signs were of a nervous or respiratory disease or of death with no premonitory signs. Gross and microscopic findings included one or more of fibrinous polyserositis, fibrinous or hemmorhagic bronchopneumonia, purulent meningitis, myocardial necrosis, focal myocarditis and valvular endocarditis. Brain, cerebrospinal fluid and lung were most reliable sites for isolation of the organism. PMID- 17422124 TI - Effects of human chorionic gonadotrophin and castration on plasma gonadal steroid hormones of the dog. AB - An intramuscular injection of 500 I.U. of human chorionic gonadotrophin resulted in an increase of plasma testosterone and progesterone concentrations in the intact male dog, but had no effect on plasma 17B-estradiol concentration. Castration caused a rapid decrease in concentration of testosterone, progesterone, and 17B-estradiol, indicating that the tests were the major organs producing these hormones in the male dog. PMID- 17422125 TI - Surgical management of premature closure of the distal ulnar growth plate in a growing dog. AB - Surgical management of deformity due to premature closure of the distal ulnar growth plate in a growing dog is discussed. The method of management selected was ulnar diaphyseal ostectomy with transphyseal wiring of the distal radius. Elbow subluxatin and degenerative joint disease as a consequence of premature ulnar closure are also discussed. PMID- 17422126 TI - Fistula between the guttural pouches and the dorsal pharyngeal recess as a sequela to guttural pouch mycosis in the horse. AB - An unusual case of a two year old gelding with severe unilateral epistaxis due to guttural pouch mycosis is reported. The lesion had spread to involve the dorso medial aspects of the right guttural pouch and had formed a fistula between the dorsal pharyngeal recess and both the left and right guttural pouches. The diagnosis, surgical treatment and postoperative management are described. PMID- 17422127 TI - Somatic cell counts in bovine milk. AB - Factors which influence somatic cell counts in bovine milk are reviewed and guidelines for their interpretation are presented. It is suggested that the thresholds of 300 000 and 250 000 cells/mL be used to identify infected quarters and cows respectively. However, it is stressed that somatic cell counts are general indicators of udder health which are subject to the influence of many factors. Therefore the evaluation of several successive counts is preferable to the interpretation of an individual count.Relationships between somatic cell counts and both milk production and milk composition are discussed. Subclinical mastitis reduces milk quality and decreases yield although the relationship between production loss and somatic cell count requires clarification. Finally the availability of somatic cell counting programs in Canada is presented. PMID- 17422128 TI - Toxocara antibodies in veterinary personnel. AB - The prevelance of antibodies to Toxocara canis was determined in 113 veterinary personnel from 22 animal clinics. One hundred and fourteen adult hospital in patients served as controls. All participants completed a questionnaire supplying personal data and specifies regarding animal exposure and dog ownership. Antibodies to Toxocara were detected by enzyme linked immunosorbent assay (ELISA) employing as antigen, excretorysecretory products from second stage larvae of Toxocara canis.Toxocara antibodies were found in ten (8.8%) of 113 veterinary workers and in 11 (9.6%) of 114 controls. Antibody prevalence was no different between males and females. Toxocara infection was not associated with dog ownership in veterinary workers or in hospitalized controls. A breakdown of the specific occupations among veterinary personnel failed to show a difference in antibody prevalence, nor did years of service as a veterinary worker correlate with the presence of Toxocara antibody.Our results suggest that toxocariasis is not associated with direct humandog contact nor with occupational exposure to dogs. PMID- 17422129 TI - Enteropathie, avec perte de proteines, secondaire a une lymphangiectasie intestinale, chez trois chiens. AB - Protein-losing enteropathy secondary to intestinal lymphangiectasia in three dogsThree cases of protein-losing enteropathy secondary to intestinal lymphangiectasia in the dog are presented.In the three cases, common clinical findings were nonresponsive diarrhea and weight loss. Subcutaneous edema was observed in case 1. Polyphagia and polydipsia were evident in cases 2 and 3.Serum examinations revealed low values of protein with a normal albumin/globulin ratio. There was histopathological evidence of dilatation of lacteals, as well as plasma cell and lymphocyte infiltration in the small intestine.In the three cases, a corticotherapy was instituted and gave good results. PMID- 17422130 TI - Disruption of the blood supply to the small colon following rectal prolapse and small colon intussusception in a mare. AB - A coliky and depressed four year old Arabian mare was examined three days after replacement of a rectal prolapse that had occurred during parturition. Fresh blood was present in the rectum and a septic peritonitis was diagnosed based upon the clinical pathological examination of the peritoneal fluid. At laparotomy the terminal 1.3 meters of small colon was found to be necrotic as a result of rupture of the terminal mesocolon and associated blood vessels. Based upon the poor prognosis the mare was euthanized. Rupture of the mesocolon was thought to be a sequel to the rectal prolapse. The anatomy predisposing the horse to these events is discussed. PMID- 17422131 TI - Vaccination guidelines questioned. PMID- 17422132 TI - Reply to dr. Wood. PMID- 17422133 TI - Pathogenesis of pemphigus vulgaris in dog and man - a review. AB - Pemphigus vulgaris is a vesiculobullous disorder that predominantly involves the oral mucous membrane of the canine and human patients. The oral lesions are usually painful erosions and ulcers. This predilection for mucous membrane may reflect the smaller number of desmosomes in the oral epithelium as compared with the epidermis. The discovery of autoantibodies against the intercellular substance of stratified epithelium in patients with pemphigus vulgaris suggests that this disease represents an autoimmune disease. The exact role of these autoantibodies in the pathogenesis of pemphigus vulgaris is not clearly understood. The diagnosis is usually based on the presence of the characteristic flaccid bullae and erosions. Biopsy specimen taken from the edge of a fresh blister and adjacent epithelial layers is suitable for routine microscopic examination and direct immunofluorescence study. Combination of corticosteroids and immunosuppresive agents are used for treatment of this disease. PMID- 17422134 TI - The Clinical and Morphological Characteristics of the Uterus of the Goat during the Period of Involution. AB - Two goats were slaughtered at zero, 24, 62 hours and five, seven, nine, 12, 15, 18, 21, 25, 28, 32 and 36 days after kidding in order to study the involutionary changes of the uterus.Uterine and vaginal discharges were varied in volume and nature and ceased before the end of day 5 postpartum. Uterine contents were found in greater volumes on days 7 to 9, in lesser volumes thereafter until no longer present on days 12 to 15 postpartum.Dimensions and weights of the genital tracts were returned to their initial nonpregnant state within a period of 28 days after kidding. PMID- 17422135 TI - Exercise in diagnostic radiology. PMID- 17422136 TI - A controversy on mechanics of sacroiliac arthrosis in the horse. PMID- 17422137 TI - Reply from dr. Rooney. PMID- 17422138 TI - Pulmonary adenomatosis (jaagsiekte) of sheep in Canada. AB - Sheep pulmonary adenomatosis has recently been reported in Canada. The literature is briefly reviewed and an account of the present status of the disease in Canada is described.Sheep pulmonary adenomatosis was first diagnosed in Canada in December 1979 in a first generation descendent of sheep imported from Great Britain. In March 1980 two further cases of sheep pulmonary adenomatosis were diagnosed in a second flock. A total of 43 sheep involving eight flocks from five provinces have been observed from December, 1979 to May, 1981. The clinical signs and laboratory findings were similar to those described in sheep from other countries. It is estimated that 30% of Canadian flocks may contain sheep imported from Great Britain during the 1970's. PMID- 17422139 TI - Coronavirus-like Particles in the Feces of a Cat with Diarrhea. AB - Coronavirus-like particles were visualized in the feces of a young domestic shorthair female cat with diarrhea. On the surface projections, these particles could be distinguished from the enteric coronavirus-like particles of human, dog, cattle and monkey origin. They appeared morphologically similar to a feline enteric coronavirus recently described by other authors. A precipitin antigen was detected in the cat feces by counterimmunoelectroosmophoresis using a rabbit antibovine coronavirus serum. PMID- 17422140 TI - A survey of mastitis in selected ontario dairy herds. AB - A mastitis survey involving 74 Ontario dairy herds was conducted. The prevalence of infection at the quarter level was found to be 4.1% with Streptococcus agalactiae, 4.5% with other streptococcal species and 8.0% with Staphylococcus aureus. Regardless of the infection status, the geometric mean somatic cell count was found to increase with age of the cow but no increase was observed with increasing stage of lactation. The percentage of cows from which a bacterial pathogen was isolated increased with age but not with stage of lactation. PMID- 17422141 TI - Combined Streptomycin-Isoniazid-Rifampin Therapy in the Treatment of Johne's Disease in a Goat. AB - Johne's disease (paratuberculosis) is an insidious, invariably fatal, chronic disease of ruminants. An increasing role for the goat as a companion animal as well as its commercial use stimulated interest in attempting to treat Johne's disease in this species. The disease tends to differ both clinically and pathologically in goats compared to cattle because the former species often has less severe intestinal involvement. It was, therefore, speculated that response to therapy may differ between cattle and goats. In addition, the combination drug regime of isoniazid, rifampin and streptomycin, widely accepted for human mycobacterial infections, has not been previously employed for treatment of Mycobacterium paratuberculosis. The clinical course and pathological findings of a case of naturally occurring Johne's disease subsequent to a combined drug regime of isoniazid, rifampin and streptomycin is discussed. PMID- 17422142 TI - Fasciola cercariae in Quebec. PMID- 17422143 TI - A study of ossification of carpal and tarsal bones in normal and hypothyroid foals. AB - The degree of ossification of carpal and tarsal bones was determined in normal foals of various ages and in hypothyroid and thyroidectomized foals. In normal foals ossification occurred very rapidly in the last few weeks of gestation and less rapidly from birth to 33 days. The ulnar carpal bone was consistently less ossified than other carpal or tarsal bones. Foals with congenital hyperplastic goitre had retarded ossification of the cuboidal bones, especially the central and third tarsal bones. Thyroidectomized foals had retarded ossification of lesser degree. PMID- 17422144 TI - Incidence of porcine stress syndrome susceptibility at the st. Cyrille record of performance station in Quebec. PMID- 17422145 TI - Intramedullary Spinal Glioblastoma in a Cat, Presented as a Cauda equina Syndrome. AB - A ten year old castrated male domestic short haired cat, was presented with clinical signs related to the cauda equina. The results of the neurological examination, diagnostic procedures, and clinical course are presented. At necropsy a glioblastoma of the caudal spinal cord was present. The difficulties in distinguishing between neoplasms arising in the cauda equina and the caudal spinal cord are discussed. A comparative discussion of the cauda equina syndrome is presented. PMID- 17422146 TI - Perforated duodenal ulcer in a cow. AB - A case report of perforated duodenal ulcer in a ten year old Holstein cow is presented. On three occasions, sudden anorexia and rapidly progressing abdominal fluid distension were associated with metabolic alkalosis, hypochloremia and hypokalemia. Rumen fluid at the time of the second episode was acidic and contained an excessive amount of chloride ion. An abdominal mass dorsal to the abomasum involving the pylorus and several loops of small bowel was identified but not corrected at surgery. Necropsy confirmed a 1.5 cm diameter duodenal ulcer 6 cm distal to the pylorus. PMID- 17422147 TI - Exercise in diagnostic radiology. PMID- 17422148 TI - Calf bacillary hemoglobinuria in Saskatchewan. PMID- 17422149 TI - Aspects epizootiologiques des affections respiratoires porcines qui sevirent au Quebec, de septembre 1980 a fevrier 1981. AB - EPIZOOTIOLOGICAL ASPECTS OF PORCINE RESPIRATORY DISEASES WHICH WERE ENCOUNTERED IN QUEBEC FROM SEPTEMBER 1980 UNTIL FEBRUARY 1981: In the spring of 1981, a questionnaire was mailed to pig farmers of Quebec, in order to determine the epizootiological factors susceptible to account for the deaths which had occurred during the period extending from September 1980 until February 1981. The mortality rate was estimated at 4.5% for that period. It proved to be higher on the farms where hogs were fattened only (5.9%), compared to those where they were raised and fattened (3.4%) and for those where the hogs were owned (6.8%) compared to those where they were fattened by contract (4.3%). During that six month period, severe mortality problems (>/= 4%) were experienced by approximately 32% of the hog producers in Quebec, mainly by those who were fattening hogs. The mortality rate was found to be proportional to the size of the herds; it proved to be definitely higher when the pigs had been bought at auction sales or from livestock dealers and even higher when the pigs had been simultaneously obtained from both sources. The results of this investigation and the importance of some epizootiological factors are discussed. The economical loss for the Province was estimated at $15 120 000 for the 126 000 pigs which died during that six month period. PMID- 17422150 TI - A canadian veterinarian overseas in the first world war. AB - In the First World War, horses were an integral part of warfare and while technological warfare was making its debut, conventional wisdom decreed that cavalry horses and pack animals had to be kept in good shape for any hope of victory. Veterinarians were important people and in the armed services they were always in short supply. One Canadian who served in the veterinary corps was William Rutledge Stubbs, the son of a veterinarian, enlisted in 1917 as a commissioned officer with the Canadian Expeditionary Force. As a captain, he served in France, in 1918. The letters he sent home during his service offer some insight into the problems faced by a young veterinarian, and in a more general way, into the life of the Canadian soldier overseas. PMID- 17422151 TI - Ischemic Neuromyopathy Associated with Steel Pellet BB Shot Aortic Obstruction in a Cat. AB - An adult female calico cat was presented with clinical signs of posterior paresis. A steel pellet was seen radiographically lodged in the caudal aorta ventral to the fourth lumbar vertebra. Aortotomy was performed and the pellet was removed. Due to the severe spinal cord damage euthanasia was performed 25 days later. A discussion of the postmortem findings and similarities between this case and thromboembolic aortic obstruction follows. PMID- 17422152 TI - Chronic copper toxicity in a dairy cow. AB - A three year old Holstein dairy cow fed a ration containing a copper supplement died of chronic copper poisoning. The concentration of copper in the liver was 331 ppm (wet weight). The typical lesions of chronic copper toxicity including icterus, hepatic fibrosis and hemoglobinemic nephrosis were found at necropsy. The chronic copper toxicity was not considered to be a herd problem since the liver copper concentration in a slaughtered cull animal and blood samples taken from five animals in the same herd were within normal limits. PMID- 17422153 TI - Un cas d'anastomose porto-cave accompagnee d'encephalopathie hepatique chez une chienne. AB - A case of portacaval shunt associated with nervous signs secondary to hepatic encephalopathy in a 18 month old Doberman bitch is reported. The dog was presented because of a noticeable weight loss over a two week period and for occasional tremor.A portacaval shunt was suspected upon the clinical observations and laboratory findings. The diagnosis was confirmed by an exploratory laparotomy.The authors discuss their findings and compare them with other cases reported in veterinary literature. PMID- 17422154 TI - Necrotic colitis in two cats - a description of the lesions. AB - The lesions in two cases of necrotic colitis in old cats are described. Both had gross lesions of a necrotic, hemorrhagic colitis without gross lesions in the small intestine. Histologically the lesions resembled those of feline panleukopenia virus infection, namely: necrosis and loss of crypt epithelium, dilation of crypts and lining of crypts by flattened epithelium, subsequent collapse of the lamina propria and hemorrhage from subepithelial capillaries. Both grossly and histologically these lesions were restricted to the colon without similar involvement of the small intestine.The histories and clinical signs, the virological and hematological studies suggest that feline panleukopenia virus was not the etiological agent in these cases. No other causal agent was identified. PMID- 17422155 TI - History of glanders and early veterinary medicine. PMID- 17422156 TI - Canine cryptococcosis associated with prolonged corticosteroid therapy. AB - A case of cryptococcosis in a five and one half year old female Irish Setter has been described. The source of the infection was not determined. This is the first documented case of canine cryptococcosis in Alberta. A review of the records of the Alberta Veterinary Services Division for the years 1961-1980 indicates this is the only laboratory case of cryptococcosis encountered in that period. PMID- 17422157 TI - Exercise in diagnostic radiology. PMID- 17422158 TI - Some orthopedic procedures in the young bovine. PMID- 17422159 TI - Reflections on modern veterinary education. AB - In this brief address, I would like to review in very broad terms some of the major quandaries facing the veterinary profession as we hurtle towards what I believe will be a very different earth in the not too distant future. Associated with these, I also wish to express some personal concerns about what we need to prepare our students for and thus fulfill our obligation to the profession at large. Finally I would like to deliver some thoughts on how we might meet these tremendous demands on our curricula. These latter reflections will by no means be all encompassing but will represent some substance for future debate. I must honestly confess that I too am anxious how best to meet several of the challenges but there must be ways and means. We need everyone's sage input and well considered opinions to develop our strategy and to come through as the elegant, humanistic, responsive and accommodating profession for which we are so well known. PMID- 17422160 TI - Porcine neonatal coccidiosis in quebec. AB - Intestinal coccidiosis was diagnosed in 110 diarrheic piglets originating from 66 farrowing operations. These piglets had been submitted alive for necropsy early after the beginning of diarrhea, and the diagnosis of coccidiosis was based on the demonstration of coccidial forms in their jejunum by histopathological examination. Enteropathogenic strains of Escherichia coli were demonstrated in only nine of the 110 piglets while the transmissible gastroenteritis virus and rotavirus were each present in three pigs. The most common lesion was a multifocal villous atrophy in the middle and lower jejunum, and ileum of the affected pigs. Coccidial forms were present in variable numbers of the cells lining the affected villi and they were mainly asexual stages (meronts and merozoites).In the affected farrowing houses diarrhea began between five and 15 days of age with the highest frequency around seven to ten days of age. Morbidity rates were very variable and mortality was less than 20%. The disease was a persistent problem in large farrowing operations on continuous farrowing programs, and losses were due mainly to retarded growth and treatment costs which were usually inefficient. The disease occurred year round with the highest prevalence in summer and fall months, and in January.Search for oocysts in fecal samples or colonic contents from 28 of the 110 pigs revealed that 13 of them were shedding oocysts identified as Isospora suis. PMID- 17422161 TI - Cecal torsion in a six month old holstein-friesian steer. AB - The clinical examination of a six month old Holstein-Friesian steer with a history of bloat and anorexia revealed a distended viscus in the right flank. At surgery, the distended viscus was found to be the cecum which was about 1.37 m (4.5 feet) long and more than 30 cm (one foot) in diameter. A counter-clockwise torsion of about 360 degrees (viewing from the right flank) involving the proximal portion of spiral colon was detected. A cecatomy was performed and a large quantity of rapeseed was removed. The torsion was reduced and the animal appeared to make an uneventful recovery but died suddenly six weeks later. PMID- 17422162 TI - Heartworm in dogs in Canada in 1981. PMID- 17422163 TI - Exercise in diagnostic radiology. PMID- 17422164 TI - Reply to "local veterinary needs poorly perceived by outsiders". PMID- 17422165 TI - Characteristics of Haemophilus pleuropneumoniae Isolates and Some Epidemiological Findings on Porcine Haemophilus Pleuropneumonia in Saskatchewan. AB - Thirty isolates of Haemophilus pleuropneumoniae from clinical and slautherhouse cases of porcine Haemophilus pleuropneumonia in Saskatchewan as well as six isolates from British Columbia and Ontario were subjected to cultural, biochemical, serological and antibiotic sensitivity tests. All strains were Gram negative pleomorphic rods or coccobacilli which grew only in the presence of V factor and all produced porphyrin from delta-aminolaevulinic acid. Biochemically, the organism was positive for urease, O-nitrophenyl-beta-D-galactopyranosidase and the fermentation of sucrose, mannitol, dextrose, lactose and xylose, but was usually negative for indole. Most strains of H. pleuropneumoniae were sensitive to chloramphenicol, furamazone, carbenicillin and ampicillin, but only about 50% were sensitive to tetracycline. Serotype 5 was more common than serotype 1 or the untyped strains among Saskatchewan isolates. In addition, serotype 3 was identified from British Columbia.Retrospective epidemiological studies showed that Haemophilus pleuropneumonia occurred and recurred on farms in the Saskatoon and adjoining districts, serviced by the diagnostic laboratories of the Western College of Veterinary Medicine and that the disease was more common among three month old pigs during the fall-winter season. PMID- 17422166 TI - Aortic valvular dysplasia with subvalvular aortic stenosis. PMID- 17422167 TI - Cost of extended open period in dairy cattle. PMID- 17422168 TI - Internal Parasites found in Exotic Birds Imported into Canada. PMID- 17422169 TI - Ethmoid hematoma of the equine. AB - Two cases of ethmoid hematoma of the equine are reported. Clinically both horses had intermittent unilateral epistaxis unassociated with exercise. In one horse, diagnosis was based on the use of an endoscope for visualization and for biopsy of a mass associated with the ethmoid turbinates. In the other horse, exploratory trephination of the posterior maxillary sinus was necessary to obtain a diagnostic biopsy specimen. Radiography was helpful in the diagnosis of one case. Surgical removal of the mass was attempted in one case. Histological examination of the biopsies was the single most definitive diagnostic technique in both cases. PMID- 17422170 TI - Cutaneous lymphosarcoma with IgG monoclonal gammopathy, serum hyperviscosity and hypercalcemia in a cat. AB - A diagnosis of cutaneous lymphosarcoma in an 11 year old castrated male Siamese cat was made. In addition to an IgG monoclonal gammopathy, a variety of unusual paraneoplastic syndromes were associated with this tumor. These included serum hyperviscosity, hypercalcemia and increased susceptibility to infection. PMID- 17422171 TI - A comparison of dichromate solution floatation and fecal smears for diagnosis of cryptosporidiosis in calves. AB - Fecal samples from eleven calves experimentally infected with cryptosporidia were examined by two methods to evaluate their sensitivity and ease of use as diagnostic techniques. Comparison of a dichromate solution floatation and fecal smear techniques indicated that the former method was more sensitive. Oocysts were detected in feces earlier and for a longer period following exposure, and were easier to visualize using the dichromate solution floatation procedure. In addition, the dichromate solution floatation technique eliminated Candida albicans which can sometimes be confused with cryptosporidial oocysts in fecal smears. Another advantage was that oocysts could readily be detected in fecal samples stored as long as 120 days in dichromate solution. Using the dichromate solution floatation technique, it was shown that shedding of cryptosporidial oocysts in feces occurred from two to 20 days after oral challenge but was not always accompanied by diarrhea. Infection with cryptosporidia was confirmed in two calves at necropsy. Calves which were not sacrificed for portmortem examination recovered without treatment. The dichromate solution floatation technique is simple, rapid, inexpensive and should facilitate detection of cryptosporidia by diagnostic laboratories and some veterinary practitioners. PMID- 17422172 TI - Vaccination guidelines for dogs and cats: 1982 update. PMID- 17422174 TI - Exercise in diagnostic radiology. PMID- 17422173 TI - Methyl alcohol toxicity in cattle. PMID- 17422175 TI - Ideas and observations concerning the present state of veterinary medicine. PMID- 17422176 TI - Vitamins a, e and selenium blood levels in the fat cow syndrome. AB - Blood plasma analyses for vitamins A, E and selenium were performed from calving to five weeks of lactation in 29 cows. Twelve of the 29 cows had fat cow syndrome. The healthy cows had significantly higher (P<0.01) plasma vitamin A (40 mug/dL) and vitamin E (5 mug/mL) levels than the cows with fat cow syndrome (29 mug vitamin A/dL and 3 mug vitamin E/mL). At parturition, vitamin A level in plasma was low (25 mug/dL) but increased progressively thereafter (up to 51 mug/dL) in healthy cows, whereas cows with fat cow syndrome had lower levels of vitamin A, bordering on deficiency. The possible role of vitamin E in the alleviation of fat cow syndrome by affecting oxidation-reduction reactions in the liver is discussed. Significant (P<0.01) difference was not observed in selenium blood plasma level (35 ng/mL) between the two groups of cows or in another random group of 12 cows clinically affected by fat cow syndrome. PMID- 17422177 TI - An encouragement to publish canadian. PMID- 17422178 TI - Diet of racing sled dogs affects erythrocyte depression by stress. AB - Fourteen racing huskies were matched into pairs then assigned to two diets, a commercial stress diet and an experimental diet. Proportions of protein: fat:carbohydrate on an available energy basis were 23:57:20 in a commercial stress diet and 28:69:3 in an experimental diet. The team participated in the 1979 Iditarod Trail race and was overtaken by an episode of diarrhea. Clinical signs were suggestive of parvovirus infection; high serum titers of parvo antibodies were found after the race. Blood examination showed normal levels of metabolites, electrolytes and enzymes after the race. Erythrocyte counts were depressed significantly during the race, by 15% in dogs fed an experimental diet and by 27% in those fed a commercial stress diet. Erythrocyte parameters have also become depressed during the racing season in middle distance sled dogs fed 28% protein (energy basis) but not 32 or 39%. Depressed red blood cell production has been demonstrated previously in dogs subjected to stress induced experimentally in several ways, and its restoration has been affected by dietary protein. Erythrocyte parameters may be useful indicies of the degree of stress in a dog as well as the adequacy of its protein intake during stress. PMID- 17422179 TI - Clinical signs and studies of the site of action of purified larkspur alkaloid, methyllycaconitine, administered parenterally to calves. AB - The purified diterpenoid alkaloid, methyllycaconitine was isolated from Delphinium brownii and injected intravenously into beef calves in order to observe the clinical signs. Following injection and depending upon dosage, calves showed a variety of signs; agitation, respiratory difficulty and loss of muscle control. Collapse occurred at higher doses of methyllycaconitine. Onset of clinical signs after injection occurred within two to three minutes, and recovery was rapid, clinical signs disappearing within five to ten minutes postinjection.The clinical signs observed are consistent with a skeletal neuromuscular site of action. Methyllycaconitine may have a curare-like action of postsynaptic blockage of nicotinic cholinergic receptors in cattle as has been demonstrated by in vitro experiments in other species. Physostigmine appears to be an effective antidote to methyllycaconitine. PMID- 17422180 TI - Lindane toxicity in pigeons. AB - A flock of Birmingham Rolles pigeons were fed a commercial whole grain pigeon feed contaminated with 2 100 ppm lindane. Soon after ingestion of the feed, symptoms including diarrhea, vomition, anorexia, depression and sudden death, consistent with a feed related organochlorine toxicity were seen. The rapeseed component of the pigeon feed contained seed which had been treated with a lindane seed dressing. Chemical analysis, history and clinical findings support a diagnosis of acute lindane toxicosis which caused sudden death in 47% of the pigeon flock. PMID- 17422181 TI - Bioavailability of chloramphenicol in cattle: variation with the number of injection sites and the concentration of the pharmaceutical preparation. AB - A bioavailability study of three commercial chloramphenicol preparations was carried out in cattle in order to determine the influence of the number of muscular injection sites and the concentration of the product on the level of chloramphenicol serum concentration. Results show that chloramphenicol should be injected at the dose of 22 mg/kg at multiple injection sites to reach 5 mug/mL of serum which is considered to be the minimal inhibitory concentration. PMID- 17422182 TI - Bovine Mastitis due to Prototheca zopfi II. AB - The occurrence of protothecosis in a dairy herd quarantined under the National Brucellosis Eradication Program is reported. Infection was detected by milk culture and the presence of serum precipitins to a culture filtrate antigen preparation of Prototheca zopfi. The alga was always cultured from the milk when serum precipitins were present. Whey antibodies were demonstrated in infected quarters. Consumption of colostrum from an infected cow may have accounted for the brief appearance of serum precipitins in young calves. A naturally infected cow was monitored for 20 months. Serum antibodies disappeared six months after lactation ended but reappeared following parturition, with both algal cells and antibodies in the colostrum. Prototheca zopfi survived a 13 month dry period. There was no spread of infection to the calf. An experimental infection of a healthy cow was short lived but the presence of both serum and whey antibodies was demonstrated. Cross-reactions between Prototheca and Brucella abortus antigens were not observed, and the association between the diseases was found to be coincidental. PMID- 17422183 TI - How does parenteral phosphorus and vitamin d(3) correct afebrile postparturient udder edema in goats? PMID- 17422184 TI - Distribution of neomycin in bull calves after intramuscular injection. AB - Neomycin sulfate was injected intramuscularly in calves. Blood and tissue samples were taken at zero, one, two, four, six, eight and 24 hours after administration. The tissues with high levels (greater than 10 mug/g) of drug at the one hour period were kidney cortex and medulla, urine, blood serum and the injection site. By 24 hours after administration only the kidney cortex and urine had high levels of neomycin. The drug could not be detected in any brain tissues and very small amounts (less than 1 mug/g) were present in the bile, thymus and vitreous humor. Levels greater than 5 mug/g were present in lung tissues for less than four hours but were greater than 2 mug/g for more than 24 hours.The mean level in the injection site was greater than 700 mug/g at one hour but only trace amounts were found at 24 hours.On the basis of the tissue drug concentration intramuscularly administered neomycin was suggested as therapeutically useful for respiratory and urinary tract infections caused by susceptible bacteria. PMID- 17422185 TI - Exercise in diagnostic radiology. PMID- 17422186 TI - Recurrent pneumothorax associated with a pulmonary emphysematous bulla in a dog: surgical correction and proposed pathogenesis. AB - A three year old dog was affected with recurrent dyspnea for two months, due to a recurrent pneumothorax. This followed an episode of mild blunt trauma to the thorax and transient strangulation. Conservative therapy of aspiration of air by repeated thoracentesis gave only temporary relief. Thoracic radiographs indicated the presence of a bulla in the right middle lung lobe. An exploratory thoracotomy confirmed the diagnosis, and a lobectomy was successfully performed. The pathogenesis of acquired pulmonary bullae is postulated. Reasons for the predisposition of the right middle lung lobe are outlined. These are related to the anatomy of the right middle bronchus, and include its position between the right pulmonary artery and vein, its oval shape, and the absence of cartilagenous support at its origin. PMID- 17422187 TI - Potential treatments for anestrus in gilts and sows. AB - Pregnant mares' serum gonadotrophin (400 IU) combined with human chorionic gonadotrophin (200 IU) was administered to anestrous gilts (n=31) and sows (n=20) in commercial herds. Two-thirds of the treated animals were mated successfully within seven days and, although no control animals were included, the response indicated that this hormone combination would be useful in herds with anestrous problems. A second experiment was conducted to evaluate the occurrence of estrus and/or ovulations in prepuberal gilts (n=eight/treatment) following injection with pregnant mares' serum gonadotrophin/human chorionic gonadotrophin or other hormones that might stimulate ovarian activity. The pregnant mares' serum gonadotrophin/human chorionic gonadotrophin combination and follicle-stimulating hormone produced estrus within ten days of injection in at least half of the treated gilts but the response was lower with gonadotrophin-releasing hormone and a prostaglandin analogue. Combinations of human chorionic gonadotrophin with small amounts of estradiol benzoate stimulated estrus and ovulation in most of the treated gilts. PMID- 17422188 TI - Feline Leprosy: A Review of Forty-four Cases from Western Canada. AB - Forty-four files of cats diagnosed as having feline leprosy were reviewed. All except one were from along the Pacific coast in the Province of British Columbia. The majority of cats were between one and three years of age and there was an increase in the number of diagnosis during the winter months.Two types of granulomatous responses similar to the tuberculoid and lepromatous phases of human leprosy were recognized. In spite of the overall similarities in the cellular response to human leprosy there were, however, significant differences such as areas of caseous necrosis and constant lack of nerve involvement. Still as there is presently no way to prove that Mycobacterium leprae is not the cause of feline leprosy the public health significance remains uncertainThe treatment of choice would appear to be surgical removal. PMID- 17422189 TI - Study of serum drug levels in calves following intramuscular administration of three tetracycline drug preparations. AB - Three antibiotic formulations, oxytetracycline (A) in propylene glycol and oxytetracycline (B) in polyvinyl pyrrolidine and pyrrolidino-methyltetracycline in an oil suspension were given to calves by the intramuscular route. Only oxytetracycline (A) appeared to cause much pain after injection.The half-time elimination (t((1/2)cl)) for oxytetracycline (A) was 14.000 +/- 3.580 h for oxytetracycline (B) 10.290 +/-5.159 h and for pyrrolidinomethyltetracycline 8.160 +/- 0.920 h. The rate of elimination ;beta slope" for oxytetracycline (A) was 0.052 +/- 0.012 h(-1) for oxytetracycline (B) 0.077 +/- 0.261 h(-1) and for pyrrolidinomethyltetracycline 0.086 +/- 0.010 h(-1). The Y intercept of the "beta" elimination slope C(os) (mug/mL) for oxytetracycline (A) was 2.490 +/- 1.040, for oxytetracycline (B) 3.463 +/- 1.874 and for pyrrolidinomethyltetracycline 2.852 +/- 1.360.Pyrrolidinomethyltetracycline appeared to have a two component elimination curve, however, only the "beta slope" was used for the calculations. PMID- 17422190 TI - Hemorrhagic Enterotoxemia Caused by Clostridium perfringens Type C in a Foal. AB - A four day old Appaloosa foal in Alberta died from hemorrhagic enterotoxemia. Beta-toxin of Clostridium perfringens was demonstrated in the intestinal contents of the foal and a pure culture of C. perfringens type C was grown from the small intestine. Histological examination showed hemorrhage and extensive necrosis of the small intestinal mucosa. Areas of necrotic tissue were surrounded by massive numbers of clostridia-like rods. There was also a moderate degree of thyroid hyperplasia. This is believed to be the first published report of hemorrhagic enterotoxemia associated with C. perfringens type C in the horse in Canada. PMID- 17422191 TI - A winter outbreak of anthrax in cattle in Saskatchewan. AB - Anthrax was diagnosed in a cow on a ranch in southwest Saskatchewan in February of 1980. Confirmation was based on bacteriological isolation from the spleen and mesenteric lymph nodes. Evidence suggests that a creek on the ranch had carried the organisms from an area of previous infection years earlier. It is noteworthy in this case that the spleen was not enlarged. Carcass disposal, vaccination, and disinfection procedures are discussed. PMID- 17422192 TI - Compendium of animal rabies vaccines marketed in Canada. PMID- 17422193 TI - Exercise in diagnostic radiology. PMID- 17422194 TI - Laboratory Diagnosis of Enteropathogenic Escherichia coli in Calves. PMID- 17422195 TI - Bovine postparturient hemoglobinuria: a review of the literature. AB - Predisposing causes of bovine post-parturient hemoglobinuria are summarized along with suspected pathogeneses, clinical signs, laboratory findings, clinical management and early experimentation. PMID- 17422196 TI - Echocardiography as a research and clinical tool in veterinary medicine. AB - Echocardiography is the accepted term for the study of cardiac ultrasound. Although a relatively new tool for the study of the heart in man it has already found wide acceptance in the area of cardiac research and in the study of clinical cardiac disease. Animals had often been used in the early experiments with cardiac ultrasound, but only recently has echocardiography been used as a research and clinical tool in veterinary medicine. In this report echocardiography is used in the research of anesthetic effects on ventricular function and clinically in the diagnosis of congestive cardiomyopathy in a cat, ventricular septal defect in a calf, and pericardial effusion in a dog. Echocardiography is now an important adjunct to the field of veterinary cardiology. PMID- 17422197 TI - Exposure to rabies - an occupational hazard for veterinarians. AB - Interviews to solicit information about animal bites and rabies vaccinations were completed on 1165 of 1175 non-military veterinarians in Illinois in 1968. Two hundred and sixty-one veterinarians reported 380 exposure incidents that precipitated the administration of rabies vaccine; 72 veterinarians had received two or more series of vaccine. Vaccine was administered after exposure from: examination of a patient (n = 230), bite (n = 79), necropsy (n = 17), other causes (n = 13) and unstated (n = 41). Eighty-six percent of the exposures were to dogs or cattle. In 231 veterinarians receiving postexposure vaccinations, where year of first vaccination and year of graduation from veterinary school were known, 97 (42%) were exposed in the six years spanning one year before graduation and four years after graduation. There were 296 veterinarians, including 12 receiving postexposure vaccination, who had received their first series of vaccine as preexposure prophylaxis. Twenty-one percent of all vaccinees (pre- and postexposure) reported reactions to the rabies vaccine. Seventy-two veterinarians reported they had been treated for an animal bite in the previous year. PMID- 17422198 TI - Clinical signs and pathology of accidental monensin poisoning in sheep. AB - The clinical signs and postmortem findings in sheep from two flocks accidentally poisoned with monensin are described. Clinical signs began within 24 hours of exposure to monensin. In the acute stages they consisted of lethargy, stiffness, muscular weakness, a stilted gait and recumbency. Feed refusal was seen in one flock but not in the second. Subacute to chronic clinical signs were decreased muscle volume of the rump and thigh. When forced to run, chronically affected sheep had a stilted, stiff legged, rocking horse gait.Gross postmortem changes were not always visible. Where visible, they affected skeletal muscles and consisted of pale streaking, with atrophy in the chronic stages. Lesions were most severe in muscles of the rump and hind limbs. Microscopically myofiber swelling and hyalinization were seen with interstitial mononuclear cell reaction and extensive sarcoplasmic mineralization in some cases. Chronic lesions consisted of fibrosis and myofiber atrophy. In lambs less than one month old, diffuse gastrointestinal hemorrhage was the only finding. PMID- 17422200 TI - Reaction to conclusions on haemophilus isolates and epidemiological findings. PMID- 17422199 TI - Une nephrose toxique chez des veaux traites par un medicament contenant des produits de degradation des tetracyclines. AB - A TOXIC NEPHROSIS IN CALVES TREATED WITH A DRUG CONTAINING TETRACYCLINE DEGRADATION PRODUCTS: Tetracyclines may prove to be toxic, especially when they are administered in excessive quantity. Toxicity and high mortality were experienced in calves that had been given the recommended dose of a drug which accidentally contained tetracycline degradation products, namely: 4 epianhydrotetracycline, epitetracycline and anhydrotetracycline. Lesions of acute and subacute nephrosis were observed. In calves dying rapidly, an abnormal coloration of skeletal muscles, myocardium and brain was also noted on postmortem examination, and a degeneration of the liver was evident. As the calves were already sick when they received the drug, dehydration and other pathogenic factors might have worsened their condition. PMID- 17422201 TI - Induction of ovulation with gonadotrophin releasing hormone and progesterone in seasonally anestrous mares. AB - Five seasonally anestrous mares were treated with a regimen of gonadotrophin releasing hormone and progesterone in an attempt to induce estrus and ovulation. The treatment induced follicular activity and estrus in all mares. Two of the five mares ovulated but none conceived. PMID- 17422202 TI - Acute anaplasmosis in imported cattle. AB - Of 18 Hereford cattle imported into Quebec from the eastern U.S.A., five exhibited acute hemolytic anemia, icterus, depression, fever, anorexia and died; 11 were killed because they had positive or suspicious anaplasma titers and two were quarantined. Anaplasma marginale organisms were found in the erythrocytes of the sick animals by light and electron microscopy. The partial absence of erythrocytic plasmalemma on several electron photomicrographs suggested exit of the anaplasma bodies. Titers up to 1:320 in infected animals were found by the complement fixation test. PMID- 17422203 TI - Exercise in diagnostic radiology. PMID- 17422204 TI - Bovine Cryptosporidiosis: Clinical and Pathological Findings in Forty-two Scouring Neonatal Calves. AB - Cryptosporidia organisms were identified in 42 of 161 (26%) neonatal, diarrheic calves, over a 32 month period commencing July 1979. Forty of the 161 calves were submitted alive and cryptosporidiosis was diagnosed in 63% (25 of 40) of them. The cryptosporidia infected calves were usually one to two weeks old and came from 26 herds where the typical history was profuse, watery diarrhea in nearly all neonatal calves. The diarrhea usually started around one week of age, was unresponsive to all conventional antidiarrhea therapies, lasted for two or more weeks and was usually fatal. Twenty-nine (69%) of the cryptosporidia infected calves were submitted between December and February. These calves were often hutch reared.Histopatholoical examination revealed large numbers of the coccidial parasite Cryptosporidium sp embedded in the microvilli of jejunal and ileal absorptive enterocytes of all affected calves. The organisms were identified as trophozoites and schizonts (asexual stages) and macrogametes (female sexual stages) with the electron microscope. Microgametes (male sexual stages) were not identified. Occasionally a merozoite (asexual stage) was also seen apparently burrowing into or about to be enveloped by a host microvillus. Observation of the organisms was much easier when diarrheic calves were submitted alive. Enterotoxigenic Escherichia coli were often cultured from intestines of dead calves and occasionally from calves submitted alive. Coronavirus particles were seen in one calf. In the last year of this study, oocysts were identified in fecal smears stained with May-Grunwald-Giemsa stain and fecal samples using a dichromate solution flotation technique. PMID- 17422205 TI - Peritoneal fluid analysis in the diagnosis of abdominal disorders in cattle: a retrospective study. AB - A retrospective study of bovine peritoneal fluids collected over a two year period was conducted. Of a total of 66 cattle studied, 31 had a nonseptic peritonitis, 11 acute bacterial peritonitis, eight ascites and 16 miscellaneous disorders such as abomasal impaction, enteritis and lymphosarcoma. Peritoneal fluid analysis was a useful aid in the diagnosis of abdominal disorders of cattle, especially as hematological changes were absent in many cases. Due to relatively low nucleated cell counts in bovine peritonitis, all parameters (i.e. nucleated cell count, total protein and differential cell counts) must be evaluated before interpretation. A nucleated cell count of greater than 6000 cells/muL and total protein content of greater than 3 g/dL was consistent with the diagnosis of peritoneal inflammation in 80% of the cases studied. An atlas of cell types common to bovine peritoneal fluid is presented. PMID- 17422206 TI - Blood chemical abnormalities in cattle with ruptured bladders and ruptured urethras. AB - A study was undertaken to determine the blood chemical variables and compare changes that occurred with ruptured bladders and ruptured urethras in steers and yearling bulls. The fatality rates were 10.5% in steers with ruptured urethras and 50.8% in steers with ruptured bladders. Both groups had significantly decreased serum sodium and chloride values and significantly increased serum creatinine, blood urea nitrogen, and plasma protein concentrations compared to the normal group. Steers with ruptured bladders were more dehydrated, had significantly lower serum sodium and chloride values and had significantly higher blood urea nitrogen concentrations than steers with ruptured urethras. The combination of a decreased serum sodium, a decreased serum chloride, and a decreased ratio of serum phosphate to blood urea nitrogen occurred consistently within the ruptured bladder group. Of the variables examined, serum phosphate was the best prognostic indicator in the steers with ruptured bladders as all of the steers that died had a serum phosphate in excess of 2.9 mmol/L. The fluid and electrolyte changes are discussed. PMID- 17422207 TI - Induction of abortion during midgestation in mares. AB - Four standardbred mares, 99 to 153 days pregnant, were treated with a synthetic prostaglandin analogue, prostalene, in an attempt to induce abortion. The mares received subcutaneous injection of either 2 mg prostalene (recommended luteolytic dose); 4 mg prostalene (double luteolytic dose) or 4 mg twice at intervals of 12 hours or 24 hours. The prostalene treatment resulted in cervical relaxation, increased tone of the uterus and decreased plasma concentrations of progesterone. None of the pregnant mares aborted within seven days after the first prostalene treatment. Abortions were subsequently induced by single or multiple intrauterine infusions using warm hypertonic saline. There were no postabortion complications. PMID- 17422208 TI - Special report on pictou cattle disease: part 1. PMID- 17422210 TI - Exercise in diagnostic radiology. PMID- 17422209 TI - Glomerular polycystic kidney disease in a dog (blue merle collie). AB - Glomerular polycystic kidney disease was diagnosed in an 11 month old, female, Blue Merle Collie. Clinical signs (polyuria, polydipsia, vomiting, diarrhea, partial anorexia) and laboratory work (blood urea nitrogen, creatinine, serum phosphorus, specific gravity, proteinuria, nonregenerative anemia) indicated chronic renal failure.However, after the study of a biopsy specimen, a definitive diagnosis was reached and the prognosis was determined. Necropsy findings and histopathological studies revealed: presence of glomerular cysts, atrophy of glomerular tufts and sclerosis of the interstitial tissue. PMID- 17422211 TI - Vaccination of Mink Against Hemorrhagic Pneumonia Using a Commercial Pseudomonas aeruginosa Bacterin. PMID- 17422212 TI - Inhibitory Action of Limulus Lysate on the Lethal Effect of Escherichia coli Endotoxin to Rabbits in vivo. PMID- 17422214 TI - Antifreeze: A Potential Danger to Dogs and Cats: CVMA Small Animal Practice Committee. PMID- 17422213 TI - Vaccination: Is it Effective in Preventing Respiratory Disease or Influencing Weight Gains in Feedlot Calves? AB - Respiratory disease, both undifferentiated and etiologically defined, remains a major problem in feedlot cattle. Vaccination has been used in an attempt to reduce the frequency and/or severity of respiratory disease in the first few weeks after the cattle arrive at the feedlot.The efficacy of vaccination has been studied both in controlled laboratory experiments and field trials as well as observational studies. (In this review, efficacy refers to the ability to reduce overall treatment rate and/or increase weight gains.) This review summarizes the data resulting from studies of vaccine efficacy.In general, there is little published data to support the use of vaccines against respiratory disease under feedlot conditions. Treatment rates and weight gains usually did not differ between vaccinated and nonvaccinated groups. The use of live bovine virus diarrhea virus vaccines was associated with a significant subsequent increase in treatment rates. Criteria to be considered in future field trials are described. PMID- 17422216 TI - Special report on pictou cattle disease: part 2. PMID- 17422215 TI - Systemic mastocytoma associated with lymposarcoma in an aged cat. AB - Systemic mastocytoma was diagnosed in a 16 year old cat. Prednisone therapy was initiated but discontinued after 11 months because of development of a duodenal ulcer. Twenty months after the initial diagnosis the cat developed dyspnea, due to enlarged pharyngeal lymph nodes. Euthanasia was performed; systemic mastocytoma and lymphosarcoma were found at necropsy. PMID- 17422217 TI - Cutaneous onchocerciasis in the horse: five cases in southwestern british columbia. AB - Five horses were presented because of a dermatitis of the forehead. Unlike previous reports, ventral midline dermatitis was not the major problem, and was present in only two of five cases. All five horses responded to levamisole therapy at a daily dosage of 5.5 g for one week. Owners were cautioned that repeat therapy may be necessary. PMID- 17422218 TI - Exercise in diagnostic radiology. PMID- 17422219 TI - Acute Enteritis in Guinea Pigs Due to Yersinia enterocolitica. PMID- 17422220 TI - Transfer and decline of maternal antibody to feline calicivirus. AB - Twelve kittens born to four queens immune to feline calicivirus acquired maternal serum neutralizing antibody to feline calicivirus primarily via the colostrum. At one week of age, their titres approached or equalled those of their dams. In the absence of feline calicivirus infection, titres of maternal antibody declined to undetected levels between ten and 14 weeks of age. The half-life of maternal antibody was approximately 15 days. PMID- 17422221 TI - Selenium levels of beef cattle in southeastern british columbia relative to supplementation and type of pasture. AB - Moderate to extremely low levels of selenium in hay were related to marginally deficient levels of selenium in sera from cattle in the Windermere Valley of southeastern British Columbia. Deficiency was most pronounced in cattle fed local hay during the winter or grazed on fertilized/irrigated pastures. Cattle on range had adequate levels of serum selenium. When sodium selenite was fed at 1.0 and 3.5 mg per head per day, serum selenium levels increased commensurate with the dose. Clinical observations indicated that a variety of clinical problems disappeared after injection or supplementation with selenium/vitamin E. PMID- 17422222 TI - Esophageal ectasia in a quarterhorse colt. AB - A one month old Quarterhorse colt was presented after a week history of bilateral nasal discharge and respiratory difficulty. The cervical esophagus was greatly dilated, tortuous and filled with diluted milk. A nasogastric tube could not be passed beyond the base of the heart. An aspiration pneumonia was found at postmortem examination and the esophageal segment from the pharynx to the base of the heart was dilated, thin-walled, had degenerative muscular changes, and a reduction in size and number of ganglion cells of the myenteric plexus. Muscular hypertrophy of the terminal esophagus had reduced its lumen size. Some similarities and disparities of this condition to achalasia of man and megaesophagus of dogs are discussed. PMID- 17422224 TI - Effective drug levels and the use of feed additive medication. PMID- 17422223 TI - The Management of Prostaglandin-controlled Breeding Programs in Beef Cattle - A Five Year Study. AB - A herd of 180 beef cows was synchronized with prostaglandin and bred by artifical insemination for five successive years. Factors influencing the success or failure of estrus synchronizing programs were identified, including the management of nutrition, postpartum interval, cow age and artificial insemination technology. Specific recommendations on the management of prostaglandin controlled breeding programs resulted in improving the pregnancy rates from 33% in year 1 to 67% in year 5. Improved management and close veterinary supervision were shown to improve the success of an estrus synchronization program. PMID- 17422225 TI - Isolation of Mycoplasma mycoides subspecies mycoides from Polyarthritis and Mastitis of Goats in Canada. AB - The clinical signs, pathomorphological changes, and microbiological findings in Canadian goats infected with Mycoplasma mycoides subspecies mycoides are discussed. The disease affected mainly young goats and was characterized by septicemia and polyarthritis. Mastitis followed by septicemia was seen in two mature goats. The diagnosis was made by culture and identification of the mycoplasma. Infected goats without clinical signs were identified by cultural and serological (complement fixation) techniques. Healthy carriers are presumably able to transmit the infection and may have brought the disease to Canada. PMID- 17422226 TI - Congenital esophagotracheal fistula as the cause of bloat in a calf. AB - A four day old Holstein calf was presented with a history of recurrent free gas bloat. An exploratory laparotomy was performed and no abnormalities were discovered in the gastrointestinal tract. An esophagotracheal fistula was suspected and confirmed by endoscopy. Surgical correction was performed by ligating the tubular connection between the trachea and esophagus and suturing closed the defects in these structures. PMID- 17422227 TI - Placentitis and Abortion in Goats and Sheep in Ontario Caused by Coxiella burnetii. PMID- 17422228 TI - Histophilus ovis. PMID- 17422229 TI - Treatment of afebrile postparturient udder edema in goats. PMID- 17422230 TI - An unusual location of foreign body in the horse. AB - A seven year old Arabian gelding was presented because of a draining tract cranial to the scapula that developed following puncture with a wooden rail four months previously and drained continuously despite antibiotic treatment.The full extent of the tract could not be determined with the horse standing but a polyethylene catheter was passed 25 cm down the tract and passed medial to the scapula. With the horse under a general anaesthetic, further probing revealed the tract was much longer. The direction, length and contents of the tract were determined by fistulograph. The tract continued medial to the scapula and terminated with two radiolucent filling defects adjacent the spinous processes of the 5th-11th thoracic vertebrae. The filling defects were located more accurately in a cranial-caudal direction by the use of radiopaque markers, and in a medial lateral direction by deep needle probing during surgery. Following surgical removal of the foreign bodies the tract healed and the horse returned to his original use. Foreign bodies and the use of fistulography are discussed. PMID- 17422231 TI - Electrocution of horses by a "hot" ground. PMID- 17422232 TI - The events of normal and abnormal postpartum reproductive endocrinology and uterine involution in dairy cows: a review. AB - The results of numerous hormonal profile studies in the dairy cow are reviewed with respect to specific hormone changes in the early postpartum period, and factors contributing to the onset of cyclic reproductive activity. Variations from the normal pattern of postpartum hormonal activity or uterine involution are reviewed also. The normal involutionary changes of the bovine uterus are discussed. PMID- 17422233 TI - Higher Chlorinated Dioxins Implicated in the Mortality of Young Pigs Kept on a Pentachlorophenol-treated Wooden Floor. AB - Young pigs raised on a pentachlorophenol-treated wooden floor showed a high mortality. The deaths ceased when the original treated wood was covered with untreated plywood. Analysis of the wood, mother's milk, and young pig tissues was carried out for pentachlorophenol and chlorinated dibenzo-p-dioxins. Pentachlorophenol was found in the wood and mother's milk but not in the young pig tissues. Concentrations of the higher chlorinated dioxins were found in the wood at the ppm level, in the mother's milk at the low ppt level and in the skin and liver of the young pigs at the ppb level. A comparison of the concentrations of hexa-and hepta-dioxin isomers in the wood, and in the piglet skin and liver indicated that a selective absorption and/or metabolism of these isomers had occurred. The results of this case history implicate the higher chlorinated dioxins in the mortality of the young pigs and underline the danger of using technical pentachlorophenol for wood treatment in association with food producing animals. PMID- 17422234 TI - Mortality rates and associated factors in equine colic operations - a retrospective study of 341 operations. AB - A retrospective survey of 300 surgical treatments for colic involving 341 interventions was carried out to determine mortality rates and associated factors. These horses had been referred to the Ontario Veterinary College over the period September 1974 to February 1980. Data from the case records was collected and stored on a computer and statistical analysis was carried out using X(2) tests.Fifty percent (150/300) of the horses survived to be discharged from the hospital. Fifty-two horses were euthanized during the operation and another ten horses should have been; if these cases are excluded the overall survival rate is 64.7% (150/232). A wide range of breeds were involved but the breed did not significantly affect survival. There was a significantly greater occurrence of serious colic in the two week to two month and one to two year age groups and significantly less in the two to four year age groups when compared with the total number of horses admitted over the same period. There was an even distribution of male and female horses but males showed a significantly lower mortality rate (57% of the males survived compared with 43% of the females). The size of the animal did not affect survival significantly. There was no seasonal variation when compared with the total number of equine patients.Survival was significantly influenced by the lesion, the preoperative packed cell volume and total plasma protein and by the length of the surgical procedure. PMID- 17422235 TI - Evaluating Risk from a 2 x 2 Table: Five Useful Measures. AB - Risk assessment from 2 x 2 tables can be very useful for both clinicians and researchers. The measures described go beyond demonstrating a statistical association between some factor and disease or death, and attempt to assess, in real world terms, just how important a particular factor may be. Five measures of risk are explained by means of a veterinary example. The odds ratio compares the odds of animals succumbing to disease or death if they are exposed to a factor, compared to the odds of succumbing if they are not exposed. Attributable rate and attributable proportion (risk) measure the amount of disease or death in the exposed group which can actually be attributed to the factor under study. Population attributable rate and population attributable proportion (risk) extend these measures to the real world: how much of the disease or death on the farm might we be able to prevent? Calculation of measures of risk from a data set requires that a number assumptions be met, and that the data used be of the highest quality. PMID- 17422236 TI - Field evaluation of a teat dip containing dodecyl benzene sulfonic Acid in preventing new mammary gland infections in a dairy herd. AB - The efficacy of a teat dip containing dodecyl benzene sulfonic acid (1.94%) in reducing the incidence of new Staphylococcus aureus infections was demonstrated in a commercial dairy herd to which a number of uninfected cows and heifers were introduced. In the first three months the occurrence of new quarter infections in the teatdipped cows was reduced by 56.49% as compared to the undipped controls. PMID- 17422237 TI - A review of the work of the health of animals branch in alberta. PMID- 17422238 TI - Exercise in diagnostic radiology. PMID- 17422239 TI - Intradermoreaction and hyposensitization in canine atopy. AB - Results obtained from allergic skin testing and hyposensitization in 81 dogs are discussed.THE POSITIVE REACTIONS ENCOUNTERED MOST FREQUENTLY WITH INTRADERMAL TESTING WERE: house dust (88.9%), cotton (59.3%), moulds (53.1%), biting insects (47.0%), feathers (37.0%), kapok (35.8%) and grasses (35.8%).Only dogs with constant and multiple sensitization were hyposensitized. The duration of treatment was eight months.Hyposensitization gave good to excellent results in 76.5% of the animals treated. Some of these patients (53.0%) required a reduced dose of corticosteroids to relieve pruritus. PMID- 17422240 TI - Controlled breeding of dairy cows with cloprostenol. AB - The percentage of dairy cows that were mated and became pregnant during a 42 day breeding period was not significantly different when animals were routinely observed for estrus and mated when detected (71%, n = 56), inseminated at estrus following a single injection of cloprostenol (72%, n = 29), or inseminated at a fixed time after two cloprostenol treatments 11 days apart (69%, n = 28). However, 11 cows in the control group were not detected and mated during the breeding period and inclusion of these animals reduced the actual pregnancy rate to 60%. Results indicated that a controlled breeding program could have practical application in dairy herds but should be used with caution. Practitioners must evaluate management programs and decide, in consultation with the herdsman, if the introduction of controlled breeding technology might be advantageous. PMID- 17422241 TI - Treatment of nutritional secondary hyperparathyroidism in the cat. PMID- 17422242 TI - Examination of the origin of increased equine serum alkaline phosphatase concentrations. AB - Serum alkaline phosphatase activity was found to be increased in 32.6% of equine samples analyzed at the Ontario Veterinary College over an 18 month period. An attempt was made using sensitivity to L-phenylalanine and heat to identify the origin of increased serum alkaline phosphatase isoenzymes present in 44 clinical cases. No difference in sensitivity to either procedure was observed for serum alkaline phosphatase from groups of foals and horses representing different clinical problems. Alkaline phosphatase of osseous tissue origin appeared to be the major source of activity for each group of animals reported. PMID- 17422243 TI - An open letter to the world veterinary congress against nuclear war. PMID- 17422244 TI - A Cytogenetic Study of Repeat-breeder Heifers and Their Embryos. AB - Twenty-three Swedish Red and White, Swedish Friesian and crossbred repeat-breeder heifers and 15 day 7 embryos produced by 11 of these heifers were subjected to cytogenetic analysis. Three heifers were found to have abnormal karyotypes; two were heterozygous for the 1/29 translocation, and one was an X-trisomy. Chromosomal anomalies which might account for embryonic death and subsequent repeat-breeding could not be detected in the embryos, however, seven out of the 15 could not be karyotyped due to the lack of cells in metaphase. The possibility of chromosomal anomalies in these embryos could not be ruled out. Three embryos produced by the heifers carrying the translocation were among those which lacked cells in mitosis. Two unfertilized ova were recovered from the X-trisomy heifer suggesting that fertilization failure rather than embryonic death was the cause of repeat-breeding. In the light of this study and similar studies in other species, it is suggested that investigations at earlier stages of development are needed. PMID- 17422246 TI - Exercise in diagnostic radiology. PMID- 17422245 TI - The effects of gonadotrophin releasing hormone administration in early postpartum dairy cows on hormone concentrations, ovarian activity and reproductive performance: a review. AB - Gonadotrophin releasing hormones have become widely used hormonal compounds in veterinary medicine, particularly with respect to bovine reproduction. The character and physiological actions of gonadotrophin releasing hormone are briefly reviewed and its clinical applications are summarized. The endocrinological research concerned with the use of gonadotrophin releasing hormone in the early postpartum period is discussed. Field trials which have been conducted to assess the effects of postpartum gonadotrophin releasing hormone administration on reproductive performance have varied widely in both design and interpretation of results. These experiments are reviewed, including the clinical trials using normal cows as well as those on cows with retained placenta. PMID- 17422247 TI - Robert Koch (1843-1910): Founder of the Science of Bacteriology and Discoverer of the Tubercle Bacillus : A Study of his Life and Work. PMID- 17422248 TI - Robert koch: centenary of the discovery of the tubercle bacillus, 1882. AB - This is an account of the life and work of Robert Koch (1843-1910), Nobel Laureate in Medicine and a founder of the science of bacteriology. In particular, Koch's researches into tuberculosis are described - the discovery of the tubercle bacillus, the controversy regarding the human and bovine types, the Koch phenomenon, and the introduction of tuberculin, which proved to be ineffective as a cure but became important as a diagnostic tool in the management of tuberculosis. By his achievements in this field, Koch may be considered to be the father of the scientific study of tuberculosis. On the occasion of the centenary of Koch's discovery of the tubercle bacillus in 1882, we pay tribute to this great German master of medicine.Robert Koch's discovery of the tubercle bacillus in 1882 was a major event in the history of medicine, a turning point in our understanding and conquest of that deadly disease which had plagued mankind for millenia. After centuries of speculation as to the possible infectious nature of tuberculosis, Koch proved conclusively that the cause of the disease was infection by a specific micro-organism which he isolated. In tuberculosis, both seed and soil play their part, but without the seed - the tubercle bacillus - there is no disease.On the occasion of the centenary of Koch's discovery of the tubercle bacillus, we pay tribute to the father of the modern scientific approach to the management of tuberculosis. PMID- 17422249 TI - Si units in veterinary medicine - 1983. PMID- 17422250 TI - Ketamine hydrochloride as an immobilizing agent for striped skunks. PMID- 17422251 TI - Experimental Production of Mastitis with Haemophilus somnus in the Lactating Bovine Mammary Gland. PMID- 17422252 TI - Transmissible gastroenteritis in Swine in Saskatchewan and some of the difficulties in diagnosing infection due to the virus. PMID- 17422253 TI - Peritoneal fluid analysis in the diagnosis of abdominal disorders in cattle: a retrospective study. PMID- 17422254 TI - Mammary hypertrophy in an ovariohysterectomized cat. AB - A four year old ovariohysterectomized domestic short-haired cat under treatment for behavioral urine spraying and idiopathic alopecia developed mammary gland hypertrophy following treatment with megestrol acetate. Withdrawal of the progestin and treatment with androgen failed to cause regression of the hypertrophy. The affected mammary gland was surgically excised and recovery was uneventful. PMID- 17422255 TI - The effect of nutrition on conception rate in beef cows at cumberland house, Saskatchewan. AB - The conception rate of cows in the Cumberland House area of Saskatchewan fed native forage grown in the area was observed to be unacceptably low. Supplementation with 12g of calcium and of phosphorus daily, and provision of cereal silage seemingly improved conception rates. It would appear that phosphorus may have been the first limiting nutrient and that a portion of the improved pregnancy rate was due to phosphorus supplementation. The low energy content of the forage, especially in years when it was badly weathered, was also judged to be a factor in conception rate in the subsequent year. It was also noted that there is a potential widespread phosphorus problem for beef cattle in Saskatchewan based on Feed Testing Service analyses of feeds. PMID- 17422256 TI - Malignant melanoma involving the aorta in a dog. AB - A four year old female Siberian husky presented with shifting hind limb lameness was euthanized and found to have disseminated malignant melanoma without oral, cutaneous or ocular involvement. The largest and possibly primary mass involved the aorta and sublumbar muscles and occluded as well as penetrated the aorta. Numerous smaller tumors were present in skeletal and soft tissues of both hind limbs, suggesting hematogenous spread from the aortic lesion. PMID- 17422257 TI - Torsion of the descending colon in a cow. AB - An acute intestinal obstruction caused by a torsion of the descending colon with incarceration and strangulation of the apex of the cecum was diagnosed in a mature Holstein cow. The clinical signs manifested were acute anorexia, depression, signs of abdominal pain, and absence of feces. Rectal examination revealed a sharp decrease in luminal size of the descending colon and taut bands at that level. The final diagnosis was obtained by exploratory celiotomy. Although surgical correction was attempted, the cow died of acute fecal peritonitis 18 hours postoperatively.Acute intestinal obstruction caused by torsion of the descending colon in the cow has not been reported in the literature. PMID- 17422258 TI - Nodular panniculitis in a female toy poodle. AB - A four month old female toy Poodle showing multiple nodules 2 to 3 cm in diameter, some of which had ruptured over the neck and trunk area, was diagnosed as having nodular panniculitis. The animal had been treated with a course of chloramphenicol and ampicillin for one month with no response. The diagnosis was based on history, clinical signs, laboratory data and dramatic response to the addition of corticosteroid to the antibiotic therapy. PMID- 17422260 TI - Exercise in diagnostic radiology. PMID- 17422259 TI - The effect of carbadox on reproduction when fed to Swine up to 60 kilograms body weight. AB - Forty female and ten male pigs were used to study the effect of carbadox on reproductive performance when included in the diet at a level of 55 mg/kg from one week of age through approximately 60 kg body weight. Five replicates, each consisting of eight females and a pair of littermate boars, all crossfostered, were used. One boar reared on medicated feed and another on an unmedicated diet, were bred to two groups of four littermates. One female in each pair had been fed the medicated diet, and the other the unmedicated diet. The same breeding scheme was carried out for two parities. The following observations were made: length of estrous cycle, weaning-to-estrus interval; conception rate; age and weight at first estrus, at breeding, at 109 days of gestation, at farrowing and after farrowing; gestation length; and litter size and weights at birth and 28 days. The results showed that carbadox had no apparent effect on reproductive performance. PMID- 17422261 TI - Charles Duncan McGilvray. PMID- 17422262 TI - Francis william schofield. PMID- 17422264 TI - Commentaries on anthrax in ruminants. PMID- 17422263 TI - Porcine rabies in Western Canada. PMID- 17422265 TI - Enzootic ataxia in lambs and kids in Saskatchewan. PMID- 17422266 TI - An epizootic of Swine influenza in ontario. AB - A swine influenza virus (H(1)N(1)) was isolated for the first time in Ontario from pigs one week to one and one-half years old during an epizootic which occurred between January and May 1981. Each herd outbreak was characterized by the sudden onset of marked respiratory distress, usually affecting the entire herd, accompanied by paroxysmal coughing, anorexia, prostration and temperatures as high as 41.5 degrees C and lasting for five to seven days. Morbidity was nearly 100%; mortality was less than 1%.Hematology, bacteriology and postmortem studies were conducted on 18 pigs from 11 farms. A lymphopenia and acute hematological inflammatory cellular responses characterized by neutrophilia with a left shift, hyperfibrinogenemia and a decreased plasma protein: fibrinogen ratio were found in 50% of the pigs. The cranial lobes of the lung were collapsed and red due to a bilateral cranioventral pneumonia which affected the cranial, middle, accessory and cranioventral aspects of the caudal lobes. Histologically, there was a necrotizing bronchitis and bronchiolitis with a neutrophilic cellular exudate. Pasteurella multocida was the species of bacterium most frequently isolated from the lung; however, mixed cultures of P. multocida frequently combined with Corynebacterium pyogenes and other species were usually identified in the lung and other organs of pigs submitted dead. PMID- 17422267 TI - Canine malignant hyperthermia: diagnosis of susceptibility in a breeding colony. AB - Fifteen related dogs were studied for susceptibility to malignant hyperthermia using halothane challenge and caffeine contracture tests. These dogs had hypertrophied muscles, were of a nervous temperament and had rectal temperatures at the upper limit of the normal range. Clinical pathology findings were mild elevations of serum aspartate transaminase and mean corpuscular hemoglobin. In vitro caffeine contracture tests were performed on muscle biopsies from five of these dogs. The concentration of caffeine required to increase resting tension by 1 g in biopsy specimens of these dogs was significantly lower than that required for control dogs: 7.6 +/- 1.38 (x +/- SEM) versus 15.5 +/- 2.52 mM (P < 0.025), and in the presence of 1% halothane, 3.6 +/- 1.44 versus 10.6 +/- 2.19 mM (P < 0.05). Internal nuclei, fiber caliber variation and fiber hypertrophy were found in histological studies of muscle biopsies. Two other dogs possibly died of a canine stress syndrome analagous to the porcine stress syndrome which occurs in malignant hyperthermia susceptible swine. Eight others of this family were anesthetized with halothane or methoxyflurane. Methoxyflurane did not trigger the syndrome. The first exposure to halothane caused death from malignant hyperthermia in two dogs and a third died on the second exposure to halothane. Postmortem findings were nonspecific. The other three dogs exposed to halothane recovered uneventfully. Inheritance of the defect conforms to a multifactorial pattern, with gradations of susceptibility. PMID- 17422268 TI - Preventive veterinary medicine in Canada: study on results of a survey. PMID- 17422269 TI - Cervical intervertebral disc protrusion in two horses. AB - Two horses with ataxia of all four limbs were found to have cervical intervertebral disc protrusion. Severe pelvic limb ataxia, proprioceptive deficits and spasticity were present in both horses with similar but less severe signs in the thoracic limbs. Cerebrospinal fluid analysis was within normal limits. Metrizamide myelography allowed definitive diagnosis in one case when a compression of the spinal cord was demonstrated at the level of the second intervertebral space. In the second case, an intervertebral disc protrusion between cervical vertebrae 6 and 7 was found at necropsy. Fiber degeneration with poor myelin staining characterized the spinal cords histologically. PMID- 17422270 TI - A strategy for control of bovine leukemia virus infection: test and corrective management. AB - Test and corrective management was applied in one dairy herd (130 milking cows) to control bovine leukemia virus infection. It consisted of: raising uninfected calves in order to establish a pool of uninfected replacement heifers, preventing transmission of bovine leukemia virus through transfer of blood from animal to animal and closing the herd. Regular herd testing was combined with selected changes in herd management. These procedures have been followed since January 1979. Prevalence of antibodies (as determined by gel-immunodiffusion) has declined markedly since the program was implemented. PMID- 17422271 TI - Responses of bulls to copper supplementation in a record of performance test station. AB - A Hereford bull from a test station, routinely treated for footrot and not responding, was further examined at the Large Animal Clinic at the Western College of Veterinary Medicine. A slight arthropathy of the right shoulder was radiographically visible. The bull was found to have a marginally low plasma copper level of 0.52 ppm. All bulls in the test station were subsequently sampled for an initial plasma copper determination. At this time 76% of the bulls at the station had plasma copper levels below 0.50 ppm. A mineral mix containing 0.2% copper was added to provide 15 ppm copper in the final ration. All bulls were sampled 28 days later with less than 1% of the bulls showing plasma copper levels below 0.50 ppm.Correlation coefficients between plasma copper levels and average daily gain were +0.067 for the presupplementation period, -0.255 for the supplementation period and -0.085 for the relationship between the change in plasma copper levels and the change in average daily gain. None of these correlations were significant (P > 0.05).During the copper supplementation period plasma copper levels increased for all breeds of bulls studied. However, there was no concomitant improvement in rate of gain by the bulls. PMID- 17422272 TI - Exercise in electrocardiographic interpretation. AB - The response of the heart to disease can be detected by assessing myocardial electrical activity. The electrocardiograph is a useful diagnostic tool which is widely available to the veterinary clinician. However, interpretation of the tracing often requires considerable experience.Regularly, electrocardiograms (ECG) of clinical cases will appear in this column. Each tracing will be preceeded by the history and the pertinent ancillary data. Interpretation and discussion of the ECG will follow. The purpose of this column is to present clinicians with examples of common electrocardiographic abnormalities. PMID- 17422273 TI - Preparation of Injectable Dantrolene for Emergency Treatment of Malignant Hyperthermia-like Syndromes. PMID- 17422275 TI - New additions to the bibliographic compilation of veterinary contributions by w. Osler. PMID- 17422274 TI - Treatment of fish furonculosis with a potentiated sulfonamide compound. PMID- 17422276 TI - A retrospective study of heart disease in doberman pinscher dogs. AB - The prevalence of gross and/or histological cardiac lesions was found to be much greater in Doberman pinscher dogs (16/26 or 62%) than in non-Doberman dogs (124/417 or 30%). At least some of the affected Dobermans were unrelated. Middle aged (mean age 4.7 yr) Dobermans of both sexes (11 M:5F) were affected. Four of the Dobermans with heart lesions had congestive cardiomyopathy; three of these four had congestive heart failure and the other one died suddenly. Prominent gross lesions were ventricular dilation and atrioventricular valvular endocardiosis. Histological lesions noted were prominent myocardial fibrosis, myofiber degeneration with fatty replacement, myofiber vacuolation and arterial intimal cushion formation. A spectrum of myocardial disease exists in Dobermans and clinically overt congestive cardiomyopathy represents one end of this spectrum. PMID- 17422277 TI - Warfarin: a review with emphasis on its use in the horse. AB - Warfarin or dicoumarol prevents the production of functional clotting factors II, VII, IX and X. Navicular disease and thrombophlebitis are examples of equine thrombotic diseases in which warfarin has been used therapeutically. The initiation of anticoagulant therapy is relatively simple but attending veterinarians must be aware of the potential risks in order to minimize them. These risks include epistaxis, bleeding into the gastrointestinal tract and at the venipuncture site, and increased susceptibility to hematoma formation following local trauma. Vitamin K, especially vitamin K(1) is a swift and specific antidote for warfarin toxicity. PMID- 17422278 TI - Hypocalcemia in a dog. AB - A case of hypocalcemia was presented with a concurrent hyperphosphatemia. Normal renal function tests excluded primary renal insufficiency. The diet was thought to contain a normal ratio of calcium and phosphorus and, therefore, nutritional secondary hyperparathyroidism was not considered. The radiographs showed normal bone density and the dog was not lame. Concentrations of immunoreactive parathyroid hormone were measured in an attempt to classify the etiology. The dog was successfully managed with calcium supplementation and vitamin D(3) or oral calcium alone. PMID- 17422279 TI - Dysuria due to urachal abscessation in calves diagnosed by contrast urography. AB - Two heifer calves with dysuria were presented to the Large Animal Clinic of the Western College of Veterinary Medicine. Although no external indications of omphalitis were present, urachal abscessation, resulting in impeded urinations, was highly suspect due to the age of the animals. Intravenous contrast urography showed abnormal bladder position in both cases and consequently afforded a presurgical diagnosis. The cases and the diagnostic techniques employed are discussed. PMID- 17422280 TI - Attempts at surgical correction of unusual colonic fistulae in the horse. AB - Two incidents of penetrating wounds into the abdominal cavity of horses are presented. In both events these had resulted in penetraton of the intestinal tract. Both animals had received only minimal veterinary attention in the acute stage and had survived with the formation of intestinal fistulae.Attempts at surgical repair resulted in failure in one animal and in closure of the fistula in the second with some subsequent cosmetic defect. PMID- 17422282 TI - Heartworm in dogs in Canada in 1982. PMID- 17422281 TI - Successful bacterin therapy in a case of chronic equine staphylococcal infection. AB - A Staphylococcus aureus infection in a mature horse, resulting from trauma, is described, which proved refractory to antibiotic therapy directed by the results of antibiotic sensitivity testing. After a prolonged course, rapid resolution of the infection was achieved with the administration of an autogenous formalized bacterin. PMID- 17422283 TI - Exercise in diagnostic radiology. PMID- 17422284 TI - Epizootic-influenza. PMID- 17422285 TI - Estrous synchronization in commercial sheep flocks in alberta and Saskatchewan. PMID- 17422286 TI - A syndrome in young beef calves (simmental cross) of unknown etiology. PMID- 17422287 TI - Possibility of virally induced lymphoma in pet ferrets. PMID- 17422288 TI - Otitis in feedlot cattle. PMID- 17422289 TI - Effects of prostaglandin f(2alpha) on corpora lutea formation and function in mated bitches. AB - Fifteen mated bitches were used to study the effects of prostaglandin F(2alpha) on ovarian endocrine function during the early and midluteal phase. Five dogs were kept as controls, five were given 250 mug/kg prostaglandin F(2alpha) twice daily between the first and fifth day of metestrus, and five were similarly treated with prostaglandin F(2alpha) between 31 and 35 days of metestrus. Function of corpora lutea was monitored by measuring serum progesterone concentrations during the first 45 days of gestation.Dogs treated with prostaglandin F(2alpha) during the early luteal phase had progesterone concentrations similar to controls and pregnancies were undisturbed in both groups. A dramatic decrease in serum progesterone concentration and abortion resulted after prostaglandin F(2alpha) administration at midpregnancy.These results indicate that prostaglandin F(2alpha) was not luteolytic during the early luteal phase and was therefore ineffective for preventing pregnancy at that time. However, at the dosage and frequency used in this study, prostaglandin F(2alpha) was luteolytic and abortifacient at midgestation. PMID- 17422290 TI - The treatment of transection of the extensor carpi radialis muscle as a result of a lacerating wound in a horse. AB - A 16 month old filly was presented with the complaint of a severe laceration to the right foreleg with resultant transection of the extensor carpi radialis. Normal principles of wound treatment were followed and a bandage and splint were used for support and immobilization. The return to partial function of the damaged extensor carpi radialis was evidenced by resolution of the wound and an improvement in the patient's gait four months after the time of injury. PMID- 17422291 TI - The duration of immunity to an inactivated adjuvanted canine parvovirus vaccine. A 52 and 64 week postvaccination challenge study. AB - Dogs were successfully isolated for a period of either 52 or 64 weeks following vaccination with an inactivated, adjuvanted canine parvovirus-2 vaccine. Antibody persisted in all ten vaccinated dogs, although in one case by 52 weeks postvaccination only virus neutralizing antibody, and not hemagglutination inhibiting antibody, could be detected. Sentinel unvaccinated dogs housed alongside the vaccinated dogs throughout the study remained free of canine parvovirus-2 antibody until challenged. Upon oral challenge with canine parvovirus-2 infected material all unvaccinated dogs developed one or more signs of canine parvovirus-2 disease, shed virus and developed antibody. None of the vaccinated dogs became overtly sick. Of the five vaccinated dogs challenged 52 weeks after vaccination, three shed virus and one showed a significant rise in antibody. At 64 weeks after vaccination only one of the five challenged dogs shed virus and showed a boost in antibody titer. PMID- 17422292 TI - Observation on shipping Fever. PMID- 17422293 TI - Mesangiocapillary Glomerulonephritis in Finn Cross-bred Lambs. PMID- 17422294 TI - Discospondylitis and orchitis associated with high Brucella titre in a dog. AB - A case of discospondylitis and orchitis associated with high Brucella titre in a dog is presented. Clinical signs included intermittent pain, poor appetite and a decreased level of physical activity. Radiographic evidence of discospondylitis was found. Histopathological findings on the testes are included. Treatment regime and clinical progress are given. Diagnosis and treatment of B. canis are described and a brief review of the treatment of discospondylitis is presented. PMID- 17422295 TI - Changes in plasma progesterone concentrations in mares treated with cloprostenol and human chorionic gonadotropin and inseminated during estrus. AB - Daily changes in the plasma progesterone concentrations were determined in eight mares treated with intramuscular injections of 250 mug cloprostenol, a prostaglandin analogue, followed five days later by 2500 I.U. human chorionic gonadotropin. A second cloprostenol injection was given 14 days after the first; the mares were then inseminated on the third and fifth day of the subsequent estrus and a second injection of human chorionic gonadotropin was administered on the fifth day. The onset of estrus following the second cloprostenol treatment was synchronized beginning three to four days after treatment in all eight mares. All eight ovulated, five mares conceived and only four foaled. Evaluation of the progesterone profiles provided reliable indicators of luteolysis, ovulation and luteal function. Decreasing plasma progesterone concentrations were associated with cloprostenol induced luteolysis or preceded spontaneous onset of estrus. The plasma progesterone concentrations increased consistently after ovulation, and in the pregnant mares, the progesterone concentrations remained high during the first month after insemination. PMID- 17422296 TI - Vaginal leiomyoma in a dog. AB - The surgical treatment of vaginal leiomyoma in a seven year old Blue Heeler bitch is described. A smooth, 12 cm diameter submucosal, intraluminal, firm mass was found on vaginal examination. It appeared to arise from the left ventral vaginal wall, cranial to the clitoris but caudal to the cervix. There was no history of urinary problems and the dog was normal in all other aspects. The treatment was surgical excision of the mass via an episiotomy. Histological examination indicated a leiomyoma. The differential diagnoses, possible etiologies and control or prevention of the condition by ovariohysterectomy are also discussed. PMID- 17422297 TI - Exercise in electrocardiographic interpretation. PMID- 17422298 TI - Early history of the animal pathology division of agriculture Canada. PMID- 17422299 TI - Answer to "observation on shipping Fever". PMID- 17422300 TI - Comparative medicine. PMID- 17422301 TI - Effet, sur quelques parametres sanguins, de l'addition de tourteau de canola CV. Tower a la ration des truies. AB - EFFECT OF THE ADDITION OF CANOLA CV. TOWER MEAL TO THE RATION OF SOWS ON SOME OF THEIR BLOOD PARAMETERS: The effect of substituting in sow diet soybean meal by canola meal (10%) and litter size on different blood constituents were studied. At day 110 of pregnancy, at weaning and at day 3 of the estrous cycle following weaning, canola meal and litter size did not affect the levels of calcium, phosphorus, magnesium, sodium, potassium, total protein and blood urea nitrogen. However, some blood parameters varied significantly among physiological stages for the same animal. Within physiological stages, large differences were observed between animals for some metabolites. PMID- 17422302 TI - Monozygous cattle twins as a result of transfer of a single embryo. AB - A single embryo obtained from a superovulated Holstein-Friesian cow resulted in female twins following transfer.Blood typing studies including 17 genetic systems showed that the twins had identical genotypes. The probability that dizygous female twins from the same parents have identical blood types by chance was calculated as 1.34 x 10(-7). PMID- 17422303 TI - Ovine Mastitis Due to Histophilus ovis. AB - The clinical, pathological and bacteriological findings related to ovine mastitis caused by Histophilus ovis are described. A high proportion of the ewes belonging to a flock were infected, but the source of the contamination could not be determined. PMID- 17422304 TI - Presence of the canine transmissible venereal tumor in the nasal cavity of dogs in the area of dakar (senegal). AB - Canine transmissible venereal tumors were observed in the nasal passages of three dogs from Dakar, Senegal. Genital tumors were not present in these dogs. These observations, combined with those of few previous reports, stress the necessity to include this neoplasm in the differential diagnosis of nasal tumors in the dog. PMID- 17422305 TI - Chronic abscesses in cats associated with an organism resembling Mycoplasma. AB - Three cats were admitted to the Ontario Veterinary College with chronic nonhealing abscesses which had not responded to a variety of local and systemic treatments.Smears made from the purulent exudate and stained with Wright's, Giemsa's and Gram's stains did not reveal the presence of any bacteria, yeast or fungal hyphae. Bacteria could not be cultured aerobically or anaerobically but organisms resembling mycoplasmas were cultured in two cases. All cats responded to tetracycline therapy. PMID- 17422306 TI - Exercise in diagnostic radiology. PMID- 17422308 TI - Clostridial myositis following ivermectin administration. PMID- 17422307 TI - Small animal section - starting a canine and feline practice. PMID- 17422309 TI - A newly recognized neurodegenerative disorder of horned hereford calves. PMID- 17422310 TI - Porcine enterovirus. PMID- 17422311 TI - Vaccination against infectious respiratory disease of cattle. PMID- 17422312 TI - Pharmaceutical legal restrictions. PMID- 17422313 TI - Pharmaceutical sales. PMID- 17422314 TI - Prostaglandin and oxytocin for synchronization of farrowing of sows. PMID- 17422315 TI - Lymphosarcoma with hypercalcemia and osteolysis in a dog. AB - Lymphosarcoma in a six year old male Doberman pinscher was accompanied by hypercalcemia, generalized osteolysis and renal calcification. Tumor involvement of bone marrow was extensive. The possible pathogenesis of hypercalcemia was thought to be the result of a locally active bone-resorbing factor secreted by the tumor cells. PMID- 17422316 TI - Mycobacterium chelonei Mastitis in a Quebec Dairy Herd. AB - An epizootic of bovine mastitis caused by a nontuberculous mycobacterial agent occurred in a large Quebec dairy herd. This mastitis problem was characterized by the occurrence of a high number of cows with severely inflamed, indurated and therapeutically incurable quarters. Routine diagnostic laboratory methods yielded negative cultural findings. Approximately 40% of the milking cows developed chronic mastitis. Poor sanitation, improper milking procedures and a faulty milking system prevailed at that time. PMID- 17422317 TI - Glycosylated hemoglobin measurement in dogs and cats: implications for its utility in diabetic monitoring. AB - The measurement of glycosylated hemoglobin (HbA(1)) levels in humans is used to indicate the degree of long-term diabetic control. Using a commercially available kit for human HbA(1), values were obtained for normal and diabetic dogs and cats. The normal range established in dogs was broad and overlapped considerably with the range in diabetics. Under the assay conditions and with a limited number of diabetic animals, the test was not found to be of value for dogs or cats. PMID- 17422318 TI - Disseminated Osteomyelitis Caused by Clostridium novyi in a Cat. AB - The history, clinical signs, laboratory and radiological findings of a cat with polyostotic osteomyelitis are described. A disseminated periosteal reaction involving the femur, humerus and tibia bilaterally was evident on radiographs. The joints were not affected. Diagnosis of osteomyelitis was confirmed by bone marrow aspiration and bone biopsy. Clostridium novyi was cultured from the bone marrow.A primary focus of infection was not identified and blood cultures were repeatedly negative. The cat was treated with procaine penicillin G, penicillin V (potassium) and metronidazole. Recovery was uneventful. PMID- 17422320 TI - Veterinarians of the north-west territories and alberta. PMID- 17422319 TI - Dermatomycosis (Ringworm) in Mule Deer (Odocoileus hemionus). AB - Six mule deer (Odocoileus hemionus) with dermatomycosis are described. Trichophyton verrucosum was isolated from four. All infections were mild and were not debilitating. The lesions involved the legs in five animals and the face in two. This is the second report of ringworm in a wild ungulate in North America. PMID- 17422321 TI - Exercise in electrocardiographic interpretation. PMID- 17422322 TI - The changes to the tax rules in the past two years. PMID- 17422323 TI - Treatment of atypical postparturient udder edema in goats. PMID- 17422324 TI - Atrial fibrillation in ten cows. AB - An irregular cardiac rhythm was identified in ten adult cows during auscultation of the heart and was subsequently characterized as atrial fibrillation by electrocardiography. The occurrence of the arrhythmia was associated with primary, organic disease of the heart in two animals which had valvular endocarditis. In seven of the other cows secondary or "functional" atrial fibrillation occurred in association with disorders of abdominal origin, six gastrointestinal disorders and one uterine torsion. Spontaneous conversion to normal sinus rhythm occurred in six cows after elimination of the primary disease. PMID- 17422325 TI - Unusual Bone Healing in a White-tailed Deer. AB - Two sequential fractures in the left hind leg of a deer and their treatments are described. The first, a tibial fracture was treated using an intramedullary pin with full cerclage wires supported by a plaster cast. This fracture healed uneventfully. A subsequent second fracture involving the distal metatarsus was stabilized by external fixation. This fracture healed with the formation of a periosteal involucrumlike structure which encased the full length of the metatarsal bone. This involucrum was thought to be due to a sub-periosteal hematoma. PMID- 17422326 TI - Transitional cell carcinoma and subsequent rupture of the canine bladder: a case report and review of the literature. AB - A transitional cell carcinoma of the bladder is reported in a ten year old male Bull Mastiff. The presenting signs were a tender, enlarged abdomen, depression and weakness. Diagnostic tests suggested a ruptured bladder. The tumor was diagnosed by exploratory laparotomy and the dog was subsequently euthanized due to the poor prognosis. A review of bladder tumors with emphasis on transitional cell carcinomas is also presented. PMID- 17422327 TI - Surgical treatment of paraphimosis in a pony. AB - Circumcision (or reefing operation) was performed on an aged pony stallion to remove excessive granulation tissue involving the preputial integument following an injury and subsequent paraphimosis. Postoperative swelling of the penis and prepuce was reduced daily with gentle massage. Initially, an improvised suspensory was used to support the penis postoperatively. The pony was able to voluntarily retract its penis six months following surgery. PMID- 17422328 TI - The effects of calving to first service interval on reproductive performance in normal cows and cows with postpartal disease. AB - The relationships between the calving to first service interval and several measures of reproductive performance were evaluated in 1738 lactation records from cows in 32 southern Ontario Holstein herds. Lactation records were divided into three mutually exclusive health categories based on the cows' postpartal disease histories.Relationships between the calving to first service interval and the first service conception rate, number of services per conception and open interval were similar for all three health categories. The first service conception rate was lower and the average number of services per conception higher in cows first bred before 60 days when compared to cows first bred after 60 days. The relationship between the calving to first service interval and the open interval indicated that for each day that breeding was delayed the open interval was extended by 0.86 days.It appeared that overall conception rates may be lower for cows first bred very early or very late, but differences in the overall conception rate were only significant for cows experiencing a reproductive tract infection. Unless very expensive semen is being used, it is suggested that disease free cows be bred at the first heat occurring after 40 days postpartum, and that cows experiencing postpartal disease be bred at the first heat occurring after 60 days postpartum. PMID- 17422329 TI - Clinical and Luteolytic Effects of Fenprostalene (A Prostaglandin F(2)alpha Analogue) in Mares. AB - A study was carried out to determine the luteolytic effect of fenprostalene, a prostaglandin F(2)alpha analogue, in mares Ten mares, that included seven cyclic mares, lactating mares and a pregnant mare were used in two experiments. In the first experiment, seven mares were treated subcutaneously with 250 mug fenprostalene and in the second experiment ten mares, including the seven mares used in the first experiment, were treated with fenprostalene and artificially inseminated during the induced estrus. Fenprostalene caused luteolysis in the normal cycling mares and the pregnant mare. Mares showed estrus within one to five days after treatment. Six of the ten mares conceived during the induced estrus and a further two conceived during the next estrus. The compound produced a side effect consisting of a small, raised, sometimes painful skin swelling at the injection site, which lasted for one to two days. PMID- 17422330 TI - Plasma progesterone concentrations in dairy cows with cystic ovaries and clinical responses following treatment with fenprostalene. AB - Sixty-two dairy cows diagnosed as having cystic ovarian degeneration were used to study the correlation between rectal palpation findings and plasma progesterone concentrations and the response of cysts to treatment using fenprostalene, a luteolytic agent. Rectal palpation accurately determined the presence of luteal cysts as confirmed by plasma progesterone concentrations of 3 ng/mL or more. Treatment with fenprostalene was very effective for luteal cysts: a high percentage of treated cows exhibited estrus within seven days after treatment. The conception rate following artifical insemination during the induced estrus was 87.5% (21/24). Rectal palpation was much less accurate for the diagnosis of follicular cysts. Cows diagnosed as having follicular cysts had wide variations in plasma progesterone concentrations. Response to fenprostalene treatment was poor in cows with nonluteinized cystic follicles associated with low progesterone concentrations. However, cows diagnosed as having follicular cysts, but with progesterone concentrations of 1 ng/mL or more, responded better to fenprostalene treatment than cows with low progesterone concentrations.It was concluded that, if correctly diagnosed, luteal cysts can be successfully treated with fenprostalene, and conception rates following treatment can be expected to be normal. PMID- 17422331 TI - Veterinarians of the north-west territories and alberta. PMID- 17422332 TI - Bovine leukosis virus infection in a dairy herd in Saskatchewan. PMID- 17422333 TI - Exercise in diagnostic radiology. PMID- 17422334 TI - Year end tax planning. PMID- 17422335 TI - A dissonant reply to "an open letter to the world veterinary congress against nuclear war". PMID- 17422336 TI - Nuclear disarmament. PMID- 17422337 TI - Effects of ethinylestradiol and danazol on embryo survival and implantation in mated beagle bitches. AB - Eleven mature Beagle bitches were artificially inseminated and then used to determine the effects of ethinylestradiol or danazol on nidation, embryo survival and maintenance of pregnancy. The dogs were grouped so that three were kept as controls, four others were treated with ethinylestradiol and the remaining four were treated with danazol. The compounds were administered orally at varying intervals after insemination. The effects of the compounds were deduced from the condition of the ovaries and uterus examined at laparotomy, and the results of whelping. Ethinylestradiol had minimal effect on embryo development and implantation. Three of four bitches treated with ethinylestradiol whelped. Similarly, danazol treatment had little or no effect on embryonic survival and implantation. It is concluded that, ethinylestradiol or danazol at the dosages and schedules used in this study did not significantly interfere with embryonic survival or implantation in pregnant Beagle bitches. PMID- 17422338 TI - Abomasal impaction of cattle in Saskatchewan. AB - Seventy-five necropsy reports of cattle diagnosed as abomasal impaction by the Pathology Department of the Western College of Veterinary Medicine were reviewed. Twenty percent of all affected animals had lesions of traumatic reticuloperitonitis and 60% were believed to be primarily dietary in origin resulting from the ingestion of too much fibre. The remaining 20% did not fit into either category. Abomasal tears, ulcers, and necrosis of the walls of the rumen, omasum, or abomasum were recorded in almost half of the cases. Pyloric stenosis was not encountered. The disease occurred mostly in winter in association with straw feed. Sixty percent of impacted beef cattle suffered from the dietary form, but the majority of dairy cattle, which would have been fed better diets, had the traumatic form which may have been precipitated by dry fibrous feeds. There was no indication of a higher incidence in heavily pregnant animals. Thirty-five of the animals necropsied had been presented alive, and the clinical features of the two main types of abomasal impaction were examined. Cattle with impactions associated with traumatic reticuloperitonitis were sick for a longer period than those animals with dietary related impactions, were single incident cases, and had a statistically significant rise in serum protein. PMID- 17422339 TI - Education needs in veterinary epidemiology in the Caribbean. AB - This paper reviews the livestock situation in the Caribbean countries including animal populations, common disease problems and some of the factors relating to disease control. It refers to the particular training needs of veterinarians that would best serve the type of epidemiological investigations required to control diseases in the area. It emphasizes the practical approach and considerations that veterinarians must exercise in dealing with the particular sociological and cultural factors of each country. Reference is also made to the problems of training and then retaining an adequate number of veterinarians in the area. PMID- 17422340 TI - Diverticulum and fistula of the lower cervical esophagus in a horse. AB - Esophageal diverticulum and perforation were diagnosed in a horse. The condition was characterized clinically by dysphagia, polypnea, diffuse swelling of the ventral aspect of neck and cutaneous fistula. Endoscopic examination revealed the presence of food material in the guttural pouch. A barium sulfate esophagram was performed to outline the diverticulum.Exploratory surgery confirmed esophageal perforation and entrapment of food material between the muscular planes of the cervical region and the horse was euthanized. At necropsy a food-filled periesophageal tract was found extending from the esophageal rupture to the left guttural pouch. PMID- 17422341 TI - Exercise in electrocardiographic interpretation. PMID- 17422342 TI - Treatment of sarcoptic mange in canadian cattle with ivermectin. AB - In 1982, approximately 13,000 Canadian cattle were treated once for sarcoptic mange with ivermectin at a dose of 10 mg per 45.4 kg of body weight. The results showed 100% efficacy against Sarcoptes scabiei and other ectoparasites present. All cattle showed marked improvement in general condition with new hair growth and apparent weight gains by 31 days posttreatment. Surveillance of treated herds for a year following treatment showed that one parenteral treatment with ivermectin eradicated the disease from closed herds. PMID- 17422343 TI - Malignant pilomatrixoma in an old english sheepdog. AB - A tumor of hair follicle origin with histological features characteristic of a benign pilomatrixoma was removed surgically from the tail of a five year old male Old English Sheepdog. The tumor recurred locally two months after surgery, and 14 months later metastatic involvement of the lungs became clinically evident. This aggressive local growth and metastasis indicate that canine pilomatrixoma, although normally considered benign, may have malignant potential which is difficult to predict histologically. PMID- 17422344 TI - Veterinarians of the north-west territories and alberta: chapter three. PMID- 17422345 TI - Campylobacter jejuni as a Cause of Abortion in Mink. PMID- 17422346 TI - Surgical removal of a granulosa cell tumor from a heifer. AB - A twelve month old, noncycling Holstein-Friesian heifer, was examined because of abnormal udder development. Rectal palpation revealed a tense right uterine horn. A solid mass in the right abdomen was detected by ballottement. A granulosa cell tumor of the right ovary was removed and nine months later the animal conceived. PMID- 17422347 TI - A Survey for the Microfilariae of the Canine Heartworm, Dirofilaria immitis, in the Calgary Region of Southern Alberta. AB - A blood survey for the prevalence of the microfilariae of the canine heartworm, Dirofilaria immitis was conducted in the Calgary area of southern Alberta between November 1977 and August 1979. A total of 514 blood samples was examined by the modified Knott's test. All of the samples were negative for D. immitis microfilariae. Wright's stained blood smears taken from 19 animals at the Calgary Zoo also proved negative. One smear from a male two-toed sloth (Choloepus didactylus) contained sheathless microfilariae which were distinguishable from those of D. immitis. These results, as well as mail survey data indicate that D. immitis is not endemic in the Calgary area.It is recommended that the modified Knott's test be used for similar large scale sampling studies. In addition, it is suggested that the testing of both native and exotic zoo animals which could serve as definitive hosts of D. immitis be continued. These animals may become local sources of infection or introduce other species of microfilariae which will have to be differentiated from those of D. immitis. PMID- 17422348 TI - The incidence and seasonal characteristics of veterinary toxicoses in Saskatchewan. AB - The species incidence and seasonal occurrence of veterinary toxicoses identified in the diagnostic toxicology laboratory at the Western College of Veterinary Medicine were tabulated during the period from 1968 to 1982. A toxicological diagnosis was indicated in 990 animals during this period of time. Lead (340 cases) and strychnine (284 cases) poisoning were the predominant toxicoses encountered. Other metal, pesticide and feed-related toxicoses were reported at a much lower incidence. A statistically significant seasonal incidence of toxicity was seen with lead, chlorinated hydrocarbon and strychnine poisoning. Strychnine (261/284 cases) and metaldehyde (12/12 cases) poisoning were seen primarily in the canine. In the bovine, a high degree of species specificity was associated with lead (294/340 cases), dicoumarol (36/36 cases) and nitrite (8/10 cases) poisoning. Copper toxicity was primarily a problem in the ovine (21/24 cases). The species specificity reflects in many instances species-specific husbandry practices, local agricultural methods or biochemical and physiological differences between species. PMID- 17422349 TI - Salmon poisoning disease in dogs on southern vancouver island. AB - Salmon poisoning disease in dogs has previously been reported in North America only along the western coast of the U.S.A. This paper presents the findings from eight affected dogs recently diagnosed on Vancouver Island, Canada. The clinical signs shown by these dogs were lethargy, anorexia, pyrexia and lymph node enlargement. The causative agent, Neorickettsia helminthoeca was observed in macrophages obtained from lymph node aspirates. This organism is transmitted to dogs in cysts of the fluke Nanophyetus salmincola salmincola within the tissues of the salmon or trout. The presence of fluke eggs in the feces of dogs showing typical signs is very suggestive of a diagnosis of salmon poisoning disease. Appropriate treatment, including chloramphenicol or oxytetracycline and fluid therapy, resulted in recovery. Prevention of salmon poisoning disease in endemic areas can be achieved by advising owners against allowing their dogs to eat raw salmon or trout. We suggest, based on the diagnoses made in these eight dogs, that Vancouver Island now be considered an endemic area for salmon poisoning disease. PMID- 17422350 TI - Pheochromocytoma in the horse and measurement of norepinephrine levels in horses. AB - Ten cases of pheochromocytoma in horses were obtained from the literature and a computer search of medical records. The clinical, laboratory and pathological features of pheochromocytoma in horses were reviewed. Pheochromocytoma is a catecholamine secreting tumor which tends to occur in older horses without breed or sex predisposition. It is usually unilateral adrenal medullary in location and benign. Malignancy was present in one horse. The most common clinical signs were sweating, tachycardia, tachypnea, muscle tremor and anxiety; however the tumor may be asymptomatic. Clinical signs were nonspecific and could be confused with other diseases, especially abdominal pain. Hyperglycemia is a consistent finding. Venous norepinephrine levels were measured in normal horses. Norepinephrine measurements may prove to be a diagnostic aid in horses with pheochromocytoma. PMID- 17422351 TI - Culicoides hypersensitivity in the horse: 15 cases in southwestern british columbia. AB - The investigation of a chronic, seasonal dermatitis of horses in southwestern British Columbia is described. Typically the history indicated an insidious onset, followed by a gradual progression in the severity of the signs each year. Lesions appeared during the warmer months of the year and tended to regress during the winter. The clinical signs consisted of areas of pruritus and excoriation, affecting predominantly the ventral midline, mane and tailhead. In all cases corticosteroid therapy relieved the pruritus and allowed the lesions to heal.The salient pathological findings were hyperkeratosis, spongiosis and a dermal infiltration of eosinophils together with mononuclear cells. These changes are typical of an allergic dermatitis, which has been recognized in many parts of the world as a hypersensitivity reaction to the bites of Culicoides spp. In this instance, the epidemiological findings relating to the geographic area, the local insect population and the distribution of lesions implicated Culicoides obsoletus as the etiological agent. PMID- 17422352 TI - Exercise in electrocardiographic interpretation. PMID- 17422353 TI - PROCEEDINGS - Food-producing Animal Research Forum: Respiratory Disease in Food Producing Animals. PMID- 17422354 TI - Veterinarians of the north-west territories and alberta. PMID- 17422355 TI - Clinical observations on an outbreak of strangles. AB - An outbreak of strangles, which occurred during the spring, summer and fall of 1980 on a Standardbred stud farm in eastern Alberta is described. The infective organism, Streptococcus equi, may have been introduced by an outside mare that was brought to the stud for breeding. All of the groups of horses on the farm were affected. For the most part, the disease was allowed to run its natural course. Only severely affected individuals were treated. During the outbreak, the foals were prophylactically treated with penicillin to prevent them from contracting the disease. Ten horses died of complications of strangles infection. PMID- 17422356 TI - Serotypes and Antimicrobial Susceptibility of Pasteurella multocida Isolated from Cattle and Pigs in Ontario. PMID- 17422358 TI - Reduction of animal suffering. PMID- 17422357 TI - Diagnosis of ovine brucellosis. PMID- 17422359 TI - Canine Distemper in Wild Raccoons (Procyon lotor) at the Metropolitan Toronto Zoo. AB - An outbreak of canine distemper, involving at least 23 wild raccoons, occurred at the Metropolitan Toronto Zoo between May and August 1981. Wild red foxes were not found with distemper and the disease did not occur in the captive collection. The main clinical signs were lethargy, bilateral purulent ocular discharge, dehydration and emaciation. The main pathological findings were generalized lymphadenopathy, splenomegaly, and interstitial pneumonia. Heavy infestations with intestinal nematodes (Baylisascaris procyonis), and unidentified cestodes were found. Cryptosporidia were found on the epithelium of the duodenum in 42% of the animals examined. Eosinophilic intracytoplasmic and intranuclear inclusions were numerous in the epithelium of the urinary bladder, renal pelvis, stomach and lung. This local outbreak was part of a major eruption of the disease in raccoons in southern Ontario which began in early 1981 and persisted until the submission of this paper in mid 1983. PMID- 17422360 TI - Clostridial cellulitis in the horse: a report of five cases. AB - Five horses with fatal clostridial cellulitis are described. The characteristic findings were the rapid development of a crepitant swelling with an associated toxemia, which in four cases followed intramuscular injections. The clinical features, diagnostic techniques and pathogenesis are discussed. The importance of an early diagnosis is emphasized. PMID- 17422361 TI - Necrotizing enteritis in a calf infected with adenovirus. AB - A three week old Holstein calf that died after a period of anorexia and diarrhea had a severe necrotizing ileitis. The lesions which were most severe above Peyer's patches contained numerous filamentous bacteria. Prominent intranuclear inclusion bodies in vascular endothelial cells in both the affected intestine and the mesenteric lymph nodes were accompanied by vasculitis and thrombosis. Adenovirus-like particles were demonstrated ultrastructurally in the inclusions.This case demonstrates a preferential association of the intestinal necrosis with the gut associated lymphoid tissue. Also, the significance of viral infection of vascular endothelium versus intestinal epithelium in determining the type and severity of injury, is deduced from this and other previously reported cases. PMID- 17422362 TI - Infectious laryngotracheitis in the niagara peninsula: a case control study. AB - The method(s) of spread of infectious laryngotracheitis in poultry flocks is uncertain. Based on an outbreak of the disease in the Niagara Peninsula in the summer of 1982, managers of infected (case) flocks and noninfected (control) flocks were interviewed to compare their management practices. The data were statistically analyzed; however, the results are not conclusive since only thirteen farms were involved in the outbreak. The results do suggest the following: farms with previous infectious laryngotracheitis infected flocks are more likely to have future outbreaks than are infectious laryngotracheitis-free farms, sanitation procedures of people entering the barns (changing boots and coveralls) appears to be related to infection status and air inlet location on the barn may be associated with infection. These items should be examined more thoroughly in future outbreaks. PMID- 17422363 TI - Therapeutic and prophylactic effects of some antibiotics on experimental pneumonic pasteurellosis. AB - Pneumonic pasteurellosis was produced in cattle seronegative for bovine herpes virus-1 and Pasteurella haemolytica using their respective aerosols four days apart. When treated with four daily intravenous oxytetracycline injections one day prior, same day as and 24 hours after P. haemolytica aerosols cattle experienced a reduced mortality. Prophylactic sustained action antibiotics given 24 hours prior to the P. haemolytica aerosol also reduced mortality, however there appeared to be a variation associated with the products used. PMID- 17422364 TI - Primary Hepatocellular Tumors in Animals Killed at Meat Packing Plants: Report of 11 cases. AB - Eight bovine, two ovine and one porcine primary hepatocellular neoplasms were found during a five year survey of tumors from meat packing plants. The tumors varied in size and usually were yellow-grey. Some were encapsulated and divided into lobules by fibrous septa. The tumor cells closely resembled normal hepatocytes and were arranged in a trabecular pattern or in sheets with caverns or were a mixture of the two. Eosinophilic intranuclear inclusions were present in one bovine and one ovine case. PMID- 17422365 TI - The physiological response of siberian husky dogs to exercise: effect of interval training. AB - Five Siberian Husky Dogs participated in an initial study to determine their physiological response to three types of exercise. Blood samples were taken prior to, and three minutes following, a 7.5 km free run and 6 km team sled run for the determination of hemoglobin, hematocrit, red and white blood cell counts, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, lactate dehydrogenase, creatine phosphokinase and serum glutamic pyruvic transaminase. Samples were also taken following a 90s sprint run. Heart rate was taken immediately after each run by palpation of the femoral pulse. Average heart rates following the 90s, 7.5 km and 6 km runs were 190 bpm, 211 bpm and 166 bpm, respectively. Mean lactate concentrations following the 90s, 7.5 km and 6 km runs were 1.74 mMol . 1(-1), 0.70 mMol . 1(-1), and 3.06 mMol . 1(-1), respectively. Elevation of lactate dehydrogenase and creatine phosphokinase was greatest following the 6 km sled run. Three of the above dogs were then studied before and after a 12 week interval training program, while three other dogs served as controls. The animals completed a three stage, submaximal treadmill test prior to and following the program. Pre and post blood samples were taken, and rectal temperature and heart rate were recorded continuously throughout the test. An analysis of variance was used to examine the significance of differences between and within groups. Although the response of heart rate, lactate, temperature and serum enzymes to submaximal exercise did not change with training significant differences between groups suggested that a more prolonged program may have resulted in such adaptations. PMID- 17422366 TI - Exercise in electrocardiographic interpretation. PMID- 17422367 TI - Veterinarians of the north-west territories and alberta: chapter five. PMID- 17422368 TI - PROCEEDINGS - Academic Program: A selection of papers and abstracts from the Academic Program of the 1983 Canadian Veterinary Medical Association Convention in Calgary. PMID- 17422369 TI - A Reevaluation of Routine Force-feeding of Dam's Colostrum to Normal Newborn Beef Calves. AB - Forty-seven beef calves born to a group of second-calf Hereford and Hereford x Angus cows were used to assess the practical value of force-feeding dam's colostrum. The first 40 calves born were assigned alternately to two equal groups (I and II). One group was force-fed up to I L of dam's colostrum per calf. All these animals were bled at 0 and 48 h after birth. A further group (III) of seven calves born were not handled until they were bled at 48 h. A variety of methods were used to estimate immunoglobulin levels in colostral whey and serum samples. In evaluating the efficiency of passive humoral antibody transfer from dam to offspring, no significant differences were evident except in radial immunodiffusion levels which were increased in group III. The percentages of calves sucking within one hour of birth were 30%, 15% and 100% for groups I, II and III, respectively. Under the conditions of this study it appears that force feeding of dam's colostrum to the newborn beef calf is disruptive and does not confer any practical benefit on such calves in terms of passive humoral antibody transfer. PMID- 17422370 TI - Infertility in a ram associated with a knobbed acrosome abnormality of the spermatozoa. AB - A yearling Rambouillet ram with an asymmetrical scrotum was examined for potential breeding soundness prior to use in a synchronized mating program in a purebred flock of 20 ewes. Initial sperm cell evaluation revealed 78% knobbed acrosomes associated with few other abnormalities of the head and midpiece. Use of the ram resulted in no conception in one group of ten synchronized ewes. One month later, the proportion of sperm cells with knobbed acrosomes was 80%. PMID- 17422371 TI - Onchocerca in horses from Western Canada and the northwestern United States: an abattoir survey of the prevalence of infection. AB - In an abattoir survey of 383 horses from the four western provinces of Canada and 240 horses from Washington, Montana and North Dakota prevalences of Onchocerca sp of 11.8% and 25.8% respectively have been demonstrated by umbilical skin specimen examinations. The results of this survey are discussed in relation to the parasite's natural history and its clinical significance in western Canada. PMID- 17422372 TI - Feline phaeohyphomycosis: treatment with ketaconazole and 5-fluorocytosine. AB - Two cats with phaeohyphomycosis, one infected with Phialophora verrucosa and the other with Exophiala jeanselmei, were treated with ketaconazole alone and in combination with 5-fluorocytosine after recurrence of the infections following surgical excision. The drugs were given orally at various doses and for various lengths of time, but were ineffective. Hepatocellular damage occurred in one cat. PMID- 17422373 TI - Tyzzer's Disease in a Puppy. PMID- 17422374 TI - The use of digitalis glycosides in the dog and cat. AB - The author reviews the pharmacological properties of digitalis glycosides and their clinical use in the control of congestive heart failure in the dog and cat. Methods of digitalization, dosage for each drug, toxic effects and drug interactions are described. PMID- 17422375 TI - Intracranial schwannoma in a cow. AB - A nine year old Hereford crossbred cow with a history of progressive neurological signs was referred to the Western College of Veterinary Medicine, Saskatoon. A large intracranial mass, histologically identified as a schwannoma, was found to be compressing the left brain stem and appeared to have arisen from the left fifth cranial nerve. PMID- 17422376 TI - Seasonal occurrence and habitat preference of rabid skunks in southern alberta. AB - Eighty cases of rabies in skunks (Mephitis mephitis) occurred in southern Alberta between December 1, 1979 and December 31, 1981. Fifty-three percent (43-80) were diagnosed during March-May (breeding and parturition). There were significantly more rabid females than males during that period. Nonrabid and rabid skunks appeared to select occupied sites rather than unoccupied sites for winter and maternity denning. PMID- 17422378 TI - Veterinarians of the north-west territories and alberta. PMID- 17422377 TI - A field trial of preshipment vaccination of calves. AB - A field trial to investigate the efficacy of vitamins ADE, a Haemophilus somnus bacterin, a pasteurella bacterin, and two intranasal infectious bovine rhinotracheitis-parainfluenza type 3 vaccines administered to beef calves at least three weeks prior to weaning and shipment was conducted.Over 1000 calves were vaccinated, but of the 692 calves shipped from the ranch of origin, only 276 calves were located in Ontario, or Quebec, feedlots. The average treatment rate was 30%. Neither vitamins ADE, H. somnus bacterin, pasteurella bacterin or the porcine tissue culture infectious bovine rhinotracheitis-parainfluenza type 3 vaccine had a significant effect on treatment rates for respiratory disease. Calves vaccinated with the temperature sensitive infectious bovine rhinotracheitis-parainfluenza type 3 vaccine had a significantly (p < 0.05) lower treatment rate than the nonvaccinated, and the porcine tissue culture infectious bovine rhinotracheitis-parainfluenza type 3 vaccinated, calves. Calves vaccinated with the temperature sensitive infectious bovine rhinotracheitis-parainfluenza type 3 vaccine did not have a significantly reduced treatment rate in comparison to nonvaccinated calves from the same source. PMID- 17422379 TI - Survey of preventive veterinary medicine in Canada: analysis of practitioners' recommendations. PMID- 17422380 TI - Nuclear disarmament. PMID- 17422381 TI - A review of cancer chemotherapy for pet animals. AB - A review of the principles of cancer chemotherapy for pet animals is presented. The various pharmacological classes of antineoplastic drugs are described with specific references to those drugs that have been widely used in veterinary medicine. PMID- 17422382 TI - The distribution of oxytetracycline in the tissues of Swine following a single oral dose. AB - A single oral dose of oxytetracycline hydrochloride (50 mg/kg) produced detectable residues in the following tissues; adrenal, bile, fat, heart, kidney (cortex), kidney (medulla), liver, lung, lymph node (mesenteric), muscle, serum, spleen, thyroid and urine. The highest residue levels were observed in the urine (441 mug/mL) at three hours after administration and they were still present at 48 hours. Maximum serum levels were observed at two hours after administration. Bile samples were positive for inhibitors in all animals sampled. Drug residues were not detected in spleen, thyroid, lymph node, adrenals and heart at 48 hours.Drug levels in important edible tissues were expressed as a percentage of drug levels in two tissues with high drug concentrations - urine and kidney cortex. The percentages were highly variable when compared with urine and much less variable when compared to kidney cortex.Kidney cortex appears to be an excellent tissue for drug residue monitoring. PMID- 17422383 TI - An evaluation of a combination of injectable anesthetic agents for use in pigs. AB - A combination of atropine sulfate, fentanyl/droperidol, ketamine hydrochloride and pentobarbital was examined for its effectiveness as an anesthesia protocol for swine weighing up to 50 kg. While the dose rate and route of administration of atropine sulfate, fentanyl/droperidol and ketamine hydrochloride was held constant, the dose rate and route of administration of pentobarbital was altered until a combination was determined which would reliably and safely produce a satisfactory level of anesthesia. Following treatment, animals were monitored and the time taken to produce an effect was recorded. The degree of anesthesia as measured by the corneal reflex and the cutaneous response to a needle prick or actual surgery and the time elapsed until signs of recovery were evident were also noted.We found a combination of atropine sulfate (0.05 mg/kg) intramuscularly and fentanyl/droperidol (1 mL/13.7 kg) intramuscularly followed by ketamine hydrochloride (11 mg/kg) intramuscularly approximately ten minutes later and 3% pentobarbital solution (9 mg/kg) intravenously two minutes after that, to be a safe and reliable method for producing surgical anesthesia of 45 minutes duration. PMID- 17422384 TI - Compendium of animal rabies vaccines marketed in Canada. PMID- 17422385 TI - Canine pemphigus vulgaris treated with gold salt therapy. AB - A nine year old spayed female Collie was diagnosed as having pemphigus vulgaris. Response to corticosteroid and antibiotic therapies was unsatisfactory. Aurothioglucose therapy was used later as the sole treatment. The dog achieved a complete remission lasting at least ten months. PMID- 17422386 TI - Testing for occult heartworm infection. AB - Heartworm infection in dogs is endemic in southern Ontario but occurs only sporadically throughout the remainder of Canada. The disease may either be associated with microfilariae in the patient's blood, a patent infection, or it may be occult. This paper describes a case of occult dirofilariasis in a dog, with emphasis on the diagnosis. A patent infection could be missed if the clinician tests an insufficient amount of blood. He should perform multiple concentration tests using either the modified Knott's technique or a filtration method. Occult infections occur in prepatent or unisexual infections, when the worms become sterile following therapy, or when the host produces antibodies that result in the destruction of the microfilariae. The recent release of a kit which detects the presence of antibodies to the adult heartworms now enables veterinarians to make an accurate diagnosis in the vast majority of dogs. PMID- 17422387 TI - An abnormal digital flexor tendon sheath in the metacarpal region in a horse. AB - An abnormal digital flexor tendon sheath was present in the mid to lower metacarpal region in a horse. Positive contrast radiography showed that it communicated with the carpal synovial sheath but not with the digital synovial sheath. Cytological and physical analysis of fluid from the abnormal tendon sheath revealed that it was compatible with normal synovial fluid. This abnormal tendon sheath was probably a congenital anomaly. PMID- 17422388 TI - Dilatation of the lower cervical esophagus in a cow. AB - Acquired megaesophagus of suspected neuromuscular origin was diagnosed in a six year old Holstein cow. The dilatation was restricted to the lower cervical esophagus. Signs included projectile regurgitation of chewed ingesta at variable periods of time after swallowing, nasal discharge of mucus and feed particles, and a cough. A secondary aspiration pneumonia was associated with this condition. The dilated portion of the esophagus was detected utilizing positive contrast radiography and fiberoptic endoscopy. Treatment consisted of feed management and antibiotics. Deglutition in the cow returned to normal over a three month period despite radiographic and fiberoptic endoscopic evidence of a persistent dilatation of the esophagus. PMID- 17422389 TI - Fecal Isolation of Corynebacterium equi from a Foal with Chronic Diarrhea. AB - Corynebacterium equi was isolated from the feces of a foal with chronic diarrhea. The foal had multiple walnut-sized masses associated with the intestine on rectal examination. These were thought to be enlarged lymph nodes and/or abscesses. The foal also had an elevated plasma fibrinogen. The diarrhea was controlled with antibiotics but recurred when the treatment was stopped. PMID- 17422390 TI - Veterinarians of the north-west territories and alberta. PMID- 17422391 TI - Specialization in practice - availability of opportunities in Canada. PMID- 17422392 TI - Veterinarians and civil defence. PMID- 17422393 TI - Nuclear war proposal. PMID- 17422394 TI - Nuclear disarmament. PMID- 17422395 TI - Arms race debate. PMID- 17422396 TI - Library collection needed. PMID- 17422397 TI - Welfare of animals in nuclear war. PMID- 17422398 TI - A moderately severe expression of classical hemophilia in a family of german shepherd dogs. AB - A kindred of German shepherd dogs with a moderately severe form of classic hemophilia was studied. The propositus was presented at five months of age because of a persistent hind limb lameness and swelling of the left stifle joint. Unclotted blood was aspirated from the swelling. A brother of this dog's dam had been diagnosed as a hemophiliac.Hemostatic tests were performed on the propositus and on ten available related dogs. On the basis of pedigree analysis, sex and specific factor VIII related activity levels in their plasmas, the dogs were classified as affected, carriers, or unaffected. The propositus and one male relative were identified as classical hemophiliacs and were classified as moderately severely affected on the basis of apparent bleeding tendency and factor VIII procoagulant levels (FVIII:C 8-10% of normal). The propositus' signs were confined to those of lameness and joint soreness and postmortem lesions were confined to a few joints (hemarthrosis). The affected male relative was asymptomatic up to the time of euthanasia (three months of age) and did not have postmortem lesions attributable to hemophilia.Although classic hemophilia (particularly in large breeds of dogs) is often a severe clinical disease, more moderate forms of the disease do occur. Such forms may be more difficult to recognize early in life. PMID- 17422399 TI - Disseminated intravascular coagulation in a horse with postpartum ulcerative colitis and laminitis. AB - Hemostatic studies were conducted on a five year old Belgian mare presented two days postpartum with colic and laminitis that was unresponsive to treatment.The mare was moderately thrombocytopenic with plasma fibrinogen levels more than twice that of a normal control horse. Platelet function as evaluated by aggregometry indicated that the circulating platelets were markedly hyporesponsive. Activated partial thromboplastin times and prothrombin times were prolonged. Para-coagulation tests (protamine sulfate and ethanol gelation) were strongly positive and fibrin degradation products were significantly elevated in the serum.The laboratory data suggested that the clinical bleeding was the result of the development of disseminated intravascular coagulation. The data were compatible with intravascular activation of the clotting mechanism, consumption of hemostatic factors, inhibition of platelet function and enhanced stimulation of the fibrinolytic mechanism.This report illustrates the complexity of the hemostatic abnormalities associated with pathological overactivation of the hemostatic mechanism. Factors such as tissue thromboplastins and/or endotoxins can stimulate disseminated intravascular coagulation, particularly during pregnancy or in the early postpartum period when a physiological "hypercoagulable" state already exists. PMID- 17422400 TI - Les differentes bandes de contention utilisees pour la confection de protheses externes chez les petits animaux. AB - THE VARIOUS ORTHOPEDIC CASTING TAPES USED FOR EXTERNAL FRACTURE FIXATION IN SMALL ANIMALS.: The most common orthopedic casting tapes which can be used for external fracture fixation in small animals are described in this paper. Their individual characteristics and indications are also discussed. PMID- 17422401 TI - Cholecystolithiasis and cholecystectomy in a dog. AB - An eight year old mixed terrier bitch was presented for examination because of apparent recurrent episodes of acute abdominal pain. Though of short duration, these episodes had been occurring for about two months. Radiographs were taken and two cholecystoliths were observed. Cholecystectomy resulted in resolution of the abdominal pain. PMID- 17422402 TI - Abces consecutifs a l'administration orale d'un aimant a une vache. AB - Following the oral administration of a magnet to a cow, a perforation of the pharyngeal mucosa occurred. The magnet underwent encapsulation in the vicinity of the stylohyoid bone. The subsequent osteitis gradually extended to the temporal bone, where another abscess developed. The cardiac signs resulting from the irritation of the vagus nerve by the first abscess and the nervous signs associated with the intracranial pressure due to the second abscess are described. PMID- 17422403 TI - Multicentric fibrosarcoma in a cat and a review of the literature. AB - A case of multicentric fibrosarcoma in a ten month old domestic short-haired cat is presented and discussed. Tumor tissue was found to involve the right distal forepaw, right shoulder area and a popliteal lymph node. This anaplastic neoplasm was concentrated primarily in subcutaneous tissues but also extended to muscle, bone and lung. The cat was found to be positive for feline leukemia virus by the ELISA test. Based on these findings, it is likely that the lesions in this case result from an interaction between the feline leukemia virus and feline sarcoma virus. PMID- 17422404 TI - Plating and grafting for nonunion of a subtrochanteric femoral fracture in a dog. AB - Healing problems following pin and wire reduction of an unstable subtrochanteric femoral fracture are described. A nonunion was treated in a two year old spayed Springer Spaniel by plating and autogenous bone grafting. PMID- 17422406 TI - Exercise in radiology. PMID- 17422405 TI - Cauda equina neuritis: a chronic idiopathic polyneuritis in two horses. AB - Two cases of cauda equina neuritis are compared and contrasted. Neurological deficits of the tail and perineum were noted and functional deficits were seen in gait, urination, defecation and cranial nerve function. Lesions consisted of nonsuppurative inflammation of the nerve trunks and proliferation of the perineurium of the cauda equina. Cranial nerve involvement in one case supported a diagnosis of polyneuritis equi rather than cauda equina neuritis. The possible etiologies and pathogenesis of this disease are discussed. PMID- 17422407 TI - Veterinarians of the north-west territories and alberta. PMID- 17422408 TI - Nuclear arms race and the peace movement. PMID- 17422409 TI - World veterinarians against nuclear war. PMID- 17422410 TI - Glycosylated hemoglobin measurement. PMID- 17422411 TI - Use of a flexible fiberoptic laparoscope as a diagnostic aid in cattle. AB - This study was performed to examine the use of a flexible fiberoptic colonoscope as a diagnostic aid in cattle. Examinations were carried out on a group of normal animals and the effects of laparoscopy on hematological parameters measured. The use of laparoscopy in the diagnosis of left displacement of the abomasum and traumatic reticuloperitonitis was assessed in two separate groups of patients. The results were compared to clinical diagnosis and exploratory laparotomy. No effect was demonstrated on the results of haematological examination in the normal cows and the procedure was concluded to be safe and innocuous. Right flank laparoscopy was not useful in detecting left displacement of the abomasum in any case in this study was not helpful in making a diagnosis. The difficulties of current methods of diagnosing left displacement of the abomasum are discussed. Laparoscopy was useful in detecting and characterizing the lesions of traumatic reticuloperitonitis and was concluded to be superior to clinical examination as a diagnostic test. PMID- 17422412 TI - Control of Sarcoptes scabiei Infestation with Ivermectin in a Large Intensive Breeding Piggery. AB - Ivermectin given in a single subcutaneous dose of 300 mug/kg was tested for safety and efficacy against swine hyperkeratotic scabies in a large breeding piggery. Approximately 15% of the adult population was showing severe clinical mange. Skin scrapings were taken from 40 affected animals to provide parasite counts. Eight of 40 animals were used as untreated controls; all test animals were injected subcutaneously on day 0 with Ivermectin. General improvement was noted as early as eight days after treatment. Scratching stopped by day 21 in all treated animals. The results are discussed in regard to a control program for Sarcoptes scabiei infestation in pigs. PMID- 17422413 TI - Guttural pouch hemorrhage associated with lesions of the maxillary artery in two horses. AB - A two year old Thoroughbred gelding, presented with guttural pouch hemorrhage, had the internal and external carotid arteries ligated. Guttural pouch mycosis was detected on endoscopic examination. After one month of topical antifungal therapy, the horse was returned and euthanized because of recurrent epistaxis. A bacterial infection of the guttural pouch with associated ulceration and hemorrhage from the maxillary artery was found at necropsy.A two year old grade gelding had ulceration and hemorrhage from the external carotid artery. Utilizing balloon-tipped catheters and arterial ligation, hemestasis was achieved in the internal carotid artery and the external carotid artery and its branches. Mycotic ulceration of the internal carotid artery was detected endoscopically and treated with local antifungal therapy for one month. Thirty-three days postoperatively the horse returned, bleeding from a lesion in the maxillary artery. The rationale for surgical intervention to prevent epistaxis associated with guttural pouch mycosis and possible etiologies for postsurgical epistaxis are discussed. PMID- 17422414 TI - Assessment of a reflectance photometer in a veterinary laboratory. AB - This report is an assessment of clinical chemistry dry reagent methodology for veterinary use. A portable reflectance photometer and dry reagent strips were used to measure canine whole blood hemoglobin, and total bilirubin, glucose, cholesterol, creatinine and urea in canine, bovine, equine and feline sera. Creatine kinase and lactate dehydrogenase were assayed in canine, bovine and equine sera. The following aspects of performance are reported: within run variation determined on canine samples, between run variation using a commercial control, correlations between dry reagent and wet reagent methodology on clinical samples, and dry reagent method serum chemistry reference values for the cow, horse and dog. A brief description of some technical advantages and limitations is included.Technical requirements were minimal while reproducibility and accuracy compared well with the wet reagent method. The dry reagent method was found to be suitable for determination of canine, bovine, equine and feline serum variables as listed above. PMID- 17422415 TI - Hyperadrenocorticism in three teddy bear hamsters. AB - Hyperadrenocorticism was diagnosed in three related teddy bear hamsters with presenting complaints of alopecia and hyperpigmentation of the skin. Treatment was attempted in two of the hamsters and was successful in one case. Metyrapone and o,p'-DDD (1,1-dichloro-2-2bis (p-chlorophenyl) ethane) were the drugs used. Necropsy and histopathological examinations revealed a pituitary chromophobe adenoma in one hamster and an adrenocortical adenocarcinoma in a second hamster. The third related hamster was clinically diagnosed as having hyperadrenocorticism but the origin of the disease has not yet been determined. PMID- 17422416 TI - A yeast related ethanol intoxication syndrome in experimental calves: prevention with nystatin. AB - In a calf nutrition-metabolism study in which a high glucose (60% glucose and 3% lard) versus a high fat (23% glucose and 30% lard) milk replacer were compared, a syndrome characterized by anorexia, an unstable gait, depression, a distended abdomen and an odor of alcohol on the breath was observed in some calves. These signs were associated with intoxicating levels of plasma ethanol and the predominance of the yeast species Torulopsis glabrata in gastrointestinal contents of afflicted calves. Nystatin, a yeast inhibitor, was extremely useful in preventing the problem. PMID- 17422417 TI - Valgus-varus deformity of the intertarsal joint in broiler chickens. AB - A lateral or medial angulation of the shaft of the distal tibiotarsal bone resulting in deviation of the lower part of the leg and frequently with bending of the proximal shaft of the tarsometatarsus is the most common leg deformity in broiler chickens. This lateral or medial deviation of the legs in broiler chickens, which has been described by many workers, deserves a specific name such as angular bone deformity or valgus-varus deformity of intertarsal joint, so that it may be separated from other varieties of lameness in broilers. PMID- 17422419 TI - Exercise in radiology. PMID- 17422418 TI - The seroprevalence of canine parvovirus-2 in a selected sample of the canine population in ontario. AB - Canine sera, collected from dogs presented to the Ontario Veterinary College between 1976 and 1980, were assessed for canine parvovirus-2 antibody using a microtitre hemagglutination-inhibition test. Special emphasis was made on the period from September 1979 to October 1980 (2892 samples). No antibody was detected in samples collected in 1976 or 1977. The first positive sera were obtained in January 1978. By the end of 1978 antibodies to canine parvovirus-2 were widespread in Ontario dogs and in 1980, 683 of 2191 dogs (31.2%) had antibody. This was before widespread vaccination was being practised and indicates canine parvovirus-2 infection occurred frequently. Evaluation of clinical records of these dogs suggested that most infections had been subclinical. PMID- 17422420 TI - Veterinarians of the north-west territories and alberta. PMID- 17422421 TI - Reversible Phenylbutazone-induced Pancytopenia in a Dog. PMID- 17422422 TI - Review of "veterinarians of the north-west territories and alberta". PMID- 17422423 TI - Retail sales tax. PMID- 17422424 TI - Animal health management and intensive livestock production. PMID- 17422425 TI - The yield of colostrum and colostral gammaglobulins in beef cows and the absorption of colostral gammaglobulins by beef calves. AB - The details of a small study on the volume of colostrum and colostral gammaglobulins produced by beef cows at the first milking immediately after calving and the serum concentrations of passively acquired colostral gammaglobulins attained by beef calves are reported. The mean volume of colostrum and colostral gammaglobulins produced were 2990 +/- 2100 (SD) mL and 314.75 +/- 211.8 (SD) g respectively. The mean serum concentrations of passively acquired colostral gammaglobulins were 29.9 +/- 10.1 (SD) ZST units (n = 40, Ranch A) and 19.2 +/- 7.5 (SD) ZST units (n = 42, Ranch B). The main reason for this difference appeared to be nutritional; the cows on ranch B being in much poorer condition and almost certainly producing less colostrum than those on ranch A. PMID- 17422426 TI - Malignant thymoma with widespread metastases in a dog: case report and brief literature review. AB - A malignant thymoma of epithelial type with metastases to the liver, spleen and bone marrow in a 16 year old female Cocker Spaniel dog is described. Lesions were considered to be incidental findings at necropsy. PMID- 17422427 TI - Comparison of swabbing and biopsy for studying the flora of the bovine uterus. AB - The objectives of this study were to evaluate the efficacy of uterine biopsy as a sampling procedure for bacteriological examination, and to assess the importance of obligate anaerobes in the bovine uterus. The aerobic and anaerobic uterine flora of cows with postpartum metritis, cows in postpartum period without metritis and repeat-breeder cows was examined by using swab and biopsy sampling techniques. Obligate anaerobes were isolated in all the 11 cows with postpartum metritis and in three of the five normal cows. No obligate anaerobes were isolated from the six repeatbreeder cows. There was a significant difference (p < 0.01) in the number of bacterial isolates obtained from samples collected by biopsy and by swabbing. A total of 72 isolates was obtained with the biopsies compared to 48 by swabbing. Obligate anaerobes make up an important part of the postpartum uterine bacterial flora, and it seems that in some instances uterine biopsy would be more satisfactory than swabbing for bacteriological examination of the uterus. PMID- 17422428 TI - Antimicrobial drug susceptibility of bacteria isolated from disease processes in cattle, horses, dogs and cats. AB - Results are presented of antimicrobial disc diffusion susceptibility testing on commonly isolated bacterial pathogens made at the Ontario Veterinary College Diagnostic Bacteriology Laboratory in 1981 and 1982. Nearly 2 000 isolates from horses, cattle, dogs and cats were tested. Comparison of resistance in the same bacterial species isolated from different animal species showed significant differences between some of the same antibiotics. PMID- 17422429 TI - Torsion of the abomasum in a one month old calf. AB - The clinical and clinicopathological findings of a one month old Holstein heifer calf presented with severe abdominal distention are reported. Preoperative evaluation and therapy were initiated and followed by an exploratory laparotomy (paracostal right flank) which revealed a 360 degrees counterclockwise torsion (viewed from the rear) of the abomasum. Severe vascular compromise of the abomasum precluded salvage of the animal. Following euthanasia and postmortem evaluation, the calf was found to have irreversible changes within the abomasum. PMID- 17422430 TI - Surgical treatment for multiple lesions of elbow dysplasia in two dogs. AB - The presence of an ununited coronoid process, an ununited anconeal process and an ectopic proximolateral radial ossification center are described in a single elbow joint of a ten month old unspayed Great Dane. This animal and a nine month old spayed Saint Bernard, also with an ununited coronoid process, both had marked degenerative osteoarthritis associated with the instabilities.Chronic lameness and significant palpable elbow joint enlargement(s) were apparent in both dogs. Radiography was used to identify the lesions and surgical treatment produced satisfactory remission of signs. PMID- 17422432 TI - Exercise in radiology. PMID- 17422431 TI - Influence de la tylosine soluble sur l'endometre de la vache. AB - THE EFFECT OF SOLUBLE TYLOSIN ON THE ENDOMETRIUM OF THE COW: Although several antibiotics are often used for intrauterine infusions, their effect on the endometrium is still unknown. An evaluation of the consequences of a single or repeated intrauterine infusion of a 1.2% solution of tylosin is reported and it is concluded that such a concentration of the antibiotic is not an irritant. PMID- 17422434 TI - Self Sufficiency in Supply of DVMs. PMID- 17422433 TI - Veterinarians of the north-west territories and alberta. PMID- 17422435 TI - Papovavirus induced feather abnormalities and skin lesions in the budgerigar: clinical and pathological findings. AB - Feather abnormalities and skin lesions caused by a papovavirus infection in budgerigars are described. Diseased one to 15 day old birds displayed a lack of nestling down feathers and filoplumes on the head and neck. Survivors older than 15 days exhibited retarded growth and temporary absence of feathers variable from bird to bird. Several birds between 15 and 25 days of age had flight feathers with total absence or marked sparseness of the vanes. After 25 days, feathers began to grow, although the tail and/or some flight feathers of some of the birds remained underdeveloped or absent for several weeks. Several of these affected birds were unable to fly and are called "runners"Microscopic lesions in the feather follicles of the affected birds less than 15 days of age, were characterized by focal, multifocal or diffuse ballooning degeneration in the lateral and axial plate cells of the barb ridges with the presence of large basophilic or amphophilic intranuclear inclusions in the same cells. Focal areas of ballooning degeneration with intranuclear inclusions were also found in the epidermis. Clinical observations made on these birds are compared with those reported in the literature for French molt. PMID- 17422436 TI - Subluxation of the carpus in thirteen horses. AB - The records of 13 horses of various breeds with subluxation of the radiocarpal, intercarpal or carpometacarpal joint, or combinations of these were reviewed. Subluxation was most common at the carpometacarpal joint (n = 10) and concomitant fractures of individual carpal bones or metacarpus II and IV were seen (n = 12).Treatment comprised of immobilization in a full leg plaster cast for from four to 18 weeks. Three animals were euthanized, two because of cast complications and one because of a request by the owner. The remaining ten were saved to function as breeding animals. PMID- 17422437 TI - Ollulanus tricuspis in an Ontario Cattery. PMID- 17422438 TI - Uveitis in the dog and cat: causes, diagnosis and treatment. AB - Uveitis is an ocular disease frequently encountered in dogs and cats. Its importance relates to the fact that it can induce severe ocular pain and or result in permanent loss of vision. In addition, uveitis can be the first or only manifestation of a systemic, life-threatening disease. Since uveitis can present in a manner similar to a number of other ocular conditions, it behooves the clinician to first establish a prompt, accurate diagnosis in order to give the appropriate prognosis and treatment.The first part of this review includes applied anatomy and physiology of the uveal tract, followed by a description of the various etiological factors found in uveitis. The second part deals with clinical signs and findings in anterior and posterior uveitis and with the sequelae of this ocular inflammatory process. Lastly, the third part contains therapeutic guidelines and a list of comprehensive reviews on this major topic in veterinary ophthalmology. PMID- 17422439 TI - Teaching assignments. PMID- 17422440 TI - Reply to specialization in practice. PMID- 17422441 TI - Salmonella dublin Septicemia in Two Puppies. AB - Two eight week old purebred female Bull Terrier puppies died within 24 hours of each other as a result of a septicemia caused by Salmonella dublin. The salient clinical features were: temperature of 41 degrees C; rapid breathing; fluid, blood-stained stools; prostration and death. Pathological findings included embolic pneumonia, splenitis, myocarditis, nephritis and meningoencephalitis. Salmonella dublin was isolated from the spleen, lung and kidneys of both puppies. PMID- 17422442 TI - The final epidemic. PMID- 17422443 TI - Routine use of protective sheaths in cattle inseminations did not improve fertility. AB - A field trial was conducted to compare 60/90 day nonreturn rate for routine inseminations with the rate obtained when an outer sheath was used to protect the insemination catheter from vaginal contamination. Eleven technicians using frozen semen from ten Holstein bulls inseminated all cows with or without the protective sheath on alternate weeks for a three month period in late winter-early spring and for a similar period in late summer-early fall. The use of protective sheaths had no effect on the nonreturn rate. PMID- 17422444 TI - Antibiotic residue testing in a beef slaughterhouse. AB - The use of the swab test on premises to screen slaughtered animals for the presence of antibiotic residues is described.The article deals with test results obtained during a 108 week period in a Toronto red meat plant. Of 487 bovine carcasses tested 12 were found positive, all of them mature cows. PMID- 17422445 TI - Torsion of abdominal testicle. PMID- 17422446 TI - Veterinarians of the north-west territories and alberta. PMID- 17422447 TI - Diagnosis of equine pheochromocytoma. PMID- 17422448 TI - Terbufos poisoning in a dairy herd. AB - This report describes the accidental poisoning of over 200 head of Holstein cattle by the organophosphate, terbufos. The ingestion of an acutely toxic dose (approximately 7.5 mg/kg) of terbufos by 84 heifers resulted in severe respiratory distress as the primary clinical sign and death within 12 hours. There was no response to treatment with atropine sulfate. One hundred and twenty milking cows received a portion of the contaminated feed diluted approximately ten times. These cattle had typical signs of organophosphate poisoning and responded to atropine sulfate. Severely affected cows received pralidoxime chloride and activated carbon 48 h after terbufos ingestion but did not respond to the drugs. Diagnosis of organophosphate poisoning was confirmed by tissue and feed analysis for terbufos and measurement of whole blood cholinesterase activity. PMID- 17422449 TI - Tracheal perforation and widespread subcutaneous emphysema in a horse. AB - A twelve year old Quarterhorse mare exhibited subcutaneous emphysema of the head, neck, upper forelimbs and thorax, and walked with a stiff gait, characterized by decreased flexion of the forelimbs. Endoscopy of the trachea revealed a small tracheal wound ninety centimeters from the external nares. The location of the lesion corresponded with a seroma present at the base of the neck. The subcutaneous emphysema and the seroma resolved without specific therapy. Blunt trauma was suggested as the cause of the tracheal lesion and subsequent subcutaneous emphysema. PMID- 17422450 TI - Retrospective evaluation of xylazine/halothane anesthesia in 125 cattle. AB - A retrospective morbidity and mortality study was carried out on 125 case records of cattle involving xylazine/halothane anesthesia at the Ontario Veterinary College, Guelph between May 1979 and February 1983.One hundred and six animals were given xylazine intramuscularly with a mean dose of 0.22 mg/kg and the remaining 19 were given a mean dose of 0.17 mg/kg intravenously. When the animals became recumbent, they were intubated and connected to an anesthetic machine which delivered halothane in oxygen.Anesthetic complications were recorded for ten cattle (8%) in the series and were mainly associated with regurgitation of ruminal contents, with or without aspiration. Three animals died as a result of the anesthetic procedure and in three other instances there were complications which were potentially fatal. Two of the anesthetic deaths were directly attributable to faulty technique rather than a specific drug response.The use of xylazine/halothane was judged to be clinically satisfactory and did not produce an unacceptable complication rate, although it is evident there is a significant risk associated with general anesthesia in ruminants. PMID- 17422451 TI - Heartworm in dogs in Canada in 1983. AB - In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously. PMID- 17422452 TI - Disseminated mastocytosis in a dog. AB - A 13 year old neutered female crossbred Poodle with disseminated mastocytosis was treated with systemic glucocorticoids and antibiotics. The diagnosis was made postmortem, based on histopathological findings. PMID- 17422453 TI - Paramphistomum spp. in Dairy Cattle in Quebec. AB - Few cases of infection with Paramphistomum spp. have been reported from cattle in Canada. During the course of a recent study of bovine fascioliasis both P. microbothrioides and P. liorchis were found in the rumen of dairy cattle slaughtered in a Quebec abattoir. Eggs in feces were distinguished on the basis of their size. Coprological analysis of 932 samples from 601 cows on 17 selected farms in Portneuf County (Quebec) revealed that 34% of the animals were infected with P. microbothrioides and 1% with P. liorchis. Based on data from one herd there appears to be significant seasonal variation in egg passage for P. microbothrioides. Furthermore, old cows exhibited a higher prevalence of infection. PMID- 17422454 TI - The use of clenbuterol for obstetrical procedures in forty cows and one horse. AB - Clenbuterol is a beta-2 sympathicomimetic drug that has been used successfully for short-term postponement of parturition in the cow and for easier manipulation of both the uterus and fetus.In a mixed practice this drug was used in 41 obstetrical cases during the 1982 and 1983 calving seasons. In 26 of 34 uterine prolapses replacement was completed without epidural anesthesia and uterine relaxation was noted in all but one case.Furthermore clenbuterol greatly facilitated the manipulation of both the uterus and the fetus while performing cesarean sections and fetotomies.Adverse reactions to the drug were not seen. PMID- 17422455 TI - Veterinarians of the north-west territories and alberta. PMID- 17422456 TI - Compendium update. PMID- 17422457 TI - Gastrosplenic torsions in sows. PMID- 17422458 TI - Our professional responsibilities relative to human-animal interactions. AB - An interesting area with great potential for benefiting and enriching the lives and conditions of people and animals is opening to us in research, service and teaching. By working with colleagues in other disciplines, we can develop new and creative ways to realize the great promise inherent in people-animal interactions properly studied and utilized.Veterinarians who understand that a strong human companion animal bond can augment people's mental and physical states will help develop sound and effective companion animal programs for individuals who are lonely or handicapped and for persons in the school systems of the community, as well as its hospices, nursing and convalescent homes, prisons and other institutions. Children experiencing the deep satisfaction of interacting with animals while young will more likely become responsible pet owners and advocates as adults. The image of the profession is enhanced when children and adults see veterinarians as concerned teachers and compassionate health professionals.We as professionals will be required not only to update our knowledge and skills, but to acquire new knowledge in fields of animal and human behavior, psychology and sociology. We are needed on interdisciplinary research teams to study human animal interactions. We will also be asked to commit time and personal energies in community programs, sometimes with no remuneration. But if skilled health professionals like veterinarians do not take the lead in establishing sound, long term companion animal programs in their own communities, everyone will suffer including the animals. How we, as individual professionals, respond will be an important reflection of our compassion and our humanity. PMID- 17422459 TI - Copper poisoning in a flock of sheep. Copper excretion patterns after treatment with molybdenum and sulfur or penicillamine. AB - During an outbreak of chronic copper poisoning, fecal and urinary copper excretion were measured following treatment with molybdenum and sulfur supplementation of the feed (0.1 g ammonium molybdate plus 1 g sodium sulfate/sheep/day) or oral penicillamine (50 mg/kg bodyweight/day) using rams in metabolism cages. Serum glutamic-oxaloacetic transaminase activities and liver levels of molybdenum and copper in sheep that died were also monitored. Within four days of starting molybdenum and sulfur supplementation a highly significant increase in fecal copper excretion was evident and the increase persisted throughout the monitoring period (five weeks - general treatment of the flock continued for another three weeks). There was no effect of the molybdenum and sulfur supplementation on urinary excretion of copper. The molybdenum and sulfur supplementation was very effective, resulting in a rapid marked decrease in mortality. Oral penicillamine treatment induced cupruresis but did not affect fecal copper excretion. The results indicated that, while the cost of penicillamine may be a limiting factor for general treatment of a flock, it may be the drug of choice for the therapy of valuable breeding animals because cupruresis may be accurately and individually controlled. Serum glutamicoxaloacetic transaminase activities were a valuable aid in diagnosing chronic copper toxicosis as well as for monitoring recovery. High initial liver copper levels were gradually reduced following molybdenum and sulfur treatment. However, at the end of the study the liver copper levels of dead sheep varied within wide limits and there were still some sheep with high liver copper levels. PMID- 17422460 TI - Rabies antibody titres in vaccinated dogs. AB - In a field study, rabies virus neutralizing antibody titres were determined by the microtest modification of the rapid fluorescent focus inhibition test before and after primary vaccination in 30 puppies, and before and after booster vaccination in 59 previously vaccinated dogs. A commercial modified live virus vaccine was used. Three weeks after primary vaccination the mean antibody titre was 102 +/- 90, but only 24 dogs presented for booster vaccination had detectable antibody levels (mean titre 12 +/- 16). The antibody responses three weeks after booster vaccination (mean 380 +/- 216) were significantly greater than the responses to primary vaccination. It was concluded that previously vaccinated dogs could have an anamnestic response to booster vaccination, even when antibodies were not detected in their sera before revaccination. PMID- 17422461 TI - Effectiveness of ivermectin paste for removal of nematodes in the horse. AB - Thirteen Standardbred horses, two to five years of age, were treated with ivermectin paste per os at 200 mug/kg of body weight and 13 were untreated. Two weeks after treatment, previously untreated horses were given the paste. Fecal samples were collected from all horses at the time of treatment and periodically thereafter up to 14 weeks and were examined for nematode eggs using the Cornell McMaster dilution and the Cornell-Wisconsin double centrifugation procedures.All horses consumed the paste readily and had no signs of toxicosis. Strongyle eggs were found in the feces of all horses before treatment but not at two to three weeks after treatment. At five to six weeks after treatment only two horses had eggs in the feces. At eight, ten, 12 and 14 weeks after treatment 27, 69, 88 and 100% of the horses examined, respectively, had a few strongyle eggs but these were no greater than 18% of that of the pretreatment samples. Ivermectin oral paste, therefore, appeared to be highly effective against both adult and immature strongyles. PMID- 17422462 TI - Tocolytic drugs for use in veterinary obstetrics. AB - THE AUTHOR PRESENTS A LITERATURE REVIEW OF TWO TOCOLYTIC AGENTS USED IN VETERINARY OBSTETRICS: isoxsuprine and clenbuterol. The medical background from which these drugs emerged for human use and to which is linked their application in animal medicine is described. Each drug is reviewed according to its pharmacology, basic considerations for its clinical use and the reports on its application in the treatment and management of obstetrical disorders in veterinary medicine. PMID- 17422463 TI - Metabolic acidosis without clinical signs of dehydration in young calves. AB - Metabolic acidosis without clinical signs of dehydration was diagnosed in four calves between nine and 21 days of age. In each calf either coma or depression with weakness and ataxia was observed. Two calves had slow deep respirations. Treatment with intravenous administration of solutions of sodium bicarbonate was accompanied by a rise in blood pH and a return to normal demeanor, ambulation and appetites, allowing these calves to return to their respective herds. PMID- 17422464 TI - William osler and comparative medicine. AB - During the last thirty years of the nineteenth century, comparative medicine deeply influenced veterinary education in Montreal, New York and Philadelphia. Of the many physicians and veterinarians involved in this movement, Sir William Osler has attracted the most biographical and historical attention. However, his contributions to comparative medicine have been characterized inexactly, partly because of his later prominence as a clinician and partly because little has been written about the history of veterinary education in Quebec.Osler's teaching and research in comparative medicine as well as his efforts to promote a veterinary profession are described and set alongside the work of other physicians and veterinarians who were his contemporaries. As a result, Osler's contributions to comparative medicine are seen to be many and important but by no means unique. Other Quebec veterinarians, including Duncan McEachran, Orphyr Bruneau, Victor T. Daubigny and J.A. Couture, and such physicians as T. Wesley Mills and J. George Adami made as many, and in some cases greater, contributions to veterinary education in Quebec than did Osler. That they have not received the degree of recognition that Osler has received reveals Osler's ability to represent values and ideals and draws attention to some essential features of late nineteenth century comparative medicine. PMID- 17422465 TI - Glycosylated hemoglobin measurement. PMID- 17422466 TI - ELISA testing for occult heartworm infection. PMID- 17422467 TI - ELISA testing for heartworm. PMID- 17422468 TI - Immunodiagnosis of occult heartworm. PMID- 17422469 TI - Testing for occult heartworm infection. PMID- 17422470 TI - Isolation of Haemophilus somnus from a Ram with Fatal Septicemia. PMID- 17422471 TI - Animal health management: magwood replies to jericho. PMID- 17422472 TI - Cerebral blastomycosis in a cat. AB - A nine year old domestic shorthair cat was presented to the Small Animal Clinic, Ontario Veterinary College, with anorexia, depression and blindness. The animal died despite treatment. At necropsy, a space occupying mass was located in the left cerebral hemisphere. Histopathologically, the mass consisted of large numbers of fungal yeast-phase cells with an associated pyogranulomatous inflammatory response. The organisms were identified as Blastomyces dermatitidis on the basis of morphology and staining characteristics.The purpose of this article is to describe the lesions of blastomycosis in the brain of a cat. PMID- 17422473 TI - Surgical treatment of laryngeal paralysis in a cat. AB - A one year old cat, with a history of respiratory dyspnea since it was eight weeks of age, was found to have abnormal function of the larynx. A castellated laryngofissure and bilateral ventriculocordectomy was performed. The respiratory distress was alleviated and the cat's personality changed from being apprehensive and elusive to that of a friendly, social animal. A slight cough remained and a fiberoptic examination four months postoperatively revealed that the right arytenoid drooped medially. Thirteen months after the arytenoid was sutured to the lateral wall of the pharynx, the cat is coughing less frequently and appears to be doing well. PMID- 17422474 TI - Surgery, using plates, nuts and bolts, for pelvic fracture reductions in two cats. AB - Pelvic fractures were repaired in two cats, following the failure of initial surgical reductions. The second surgeries in both animals involved the application of bone plates using nuts and bolts. A lateral plate was used to replace wire for cat I and a ventral plate was used to spread the pelvis in the second cat after screws had failed to hold a plate on the ilium. PMID- 17422475 TI - Effect of dosage of cloprostenol on induction of farrowing and body temperature of sows. AB - In an experiment involving 161 farrowings, cloprostenol was injected on day 112 or 113 of gestation at the recommended dosage (175 mug) or a lower dosage (125 mug). Cloprostenol treatment did not result in abnormally high body temperatures of sows at parturition. Farrowing began within 29 hours of injection in 94% and 88% of the sows treated with 175 mug and 125 mug cloprostenol respectively, as compared to 15% of saline-injected controls. The duration of farrowing and number stillborn were not affected by treatment. Sows farrowing within 19 hours of treatment tended to have a large number of piglets and a higher body temperature postpartum. PMID- 17422476 TI - An Outbreak of Cysticercus ovis in Nova Scotia. AB - In 1980 and 1981, Cysticercus ovis infestations were found in lambs at slaughter from four geographically separated farms in Nova Scotia. Epidemiological investigations showed that a common factor was the presence of dogs which came from or had been trained on a fifth farm. It was subsequently shown that sheep on this latter farm were also infested.Not allowing dogs access to raw mutton, offal and sheep carcasses and the regular treatment of dogs for tapeworms controlled the outbreaks on the four farms. PMID- 17422477 TI - Un osteosarcome de la diaphyse du femur d'un chien, associe a un implant metallique. AB - AN OSTEOSARCOMA OF THE FEMORAL DIAPHYSIS ASSOCIATED WITH THE PRESENCE OF A BONE PLATE IN A DOG: A bone tumor which developed in the middle third of the left femur of an Irish Setter, seven years after a metallic plate had been used to reduce a fracture, is described. Through radiographs, the tumor was found to involve the diaphysis of the bone. The histopathology revealed that it was an osteosarcoma. Spontaneous and induced osteosarcomas are compared, and the possible correlation between the development of a tumor of this nature and the use of orthopedic implants is discussed. PMID- 17422478 TI - The cattle trade of Western Canada. PMID- 17422480 TI - Diagnosis of Corynebacterium equi Diarrhea - a Reply. PMID- 17422479 TI - Grads without jobs. PMID- 17422481 TI - Diagnosis of Corynebacterium equi Diarrhea. PMID- 17422482 TI - Feline mammary carcinoma: a retrospective evaluation of 17 cases. AB - Seventeen biopsies of feline mammary carcinoma submitted to the Veterinary Pathology Laboratory, Nova Scotia Department of Agriculture and Marketing were reviewed. All 17 cases were female cats. Data on age, reproductive status (sexually intact vs. neutered), therapy, outcome of the cases and histological features were consistent with data on feline mammary carcinoma previously reported. Four of these 17 cats had a history of receiving exogenous progestin prior to tumor development. The possible role of progestins as initiators or promoters of feline mammary carcinoma was discussed. The use of feline mammary carcinoma as a model for carcinoma of the breast in women was reviewed. PMID- 17422483 TI - Torsion of abdominal organs in sows: a report of 36 cases. AB - Necropsy records of 36 sows with torsion of abdominal organs involving individually the stomach, the spleen, a liver lobe or the intestine were reviewed for the years 1970 to 1983, and the age, the clinical signs and the gross lesions were recorded. These acute abdominal accidents were characterized clinically by sudden death. Dry sows from large breeding units were affected. Twenty-six cases were diagnosed between January 1981 and December 1983 while only ten cases had been seen between 1970 and 1980. Gastric torsion was the most common condition (40% of the cases) and the other three conditions were equally represented (20% each). Management practises that could be responsible for the apparent increase in occurrence of this problem are discussed. PMID- 17422484 TI - Therapy of feline urethral obstruction. PMID- 17422485 TI - Acetaminophen toxicosis in a cat. AB - A seven month old domestic shorthaired male cat was presented with a known history of acetaminophen ingestion. Clinical findings included icterus, depression, hypothermia, tachypnea and pronounced edema of the head and neck. Treatment was aimed at providing substrate to assist in conjugation of the drug and reversing methemoglobinemia. Administration of oral acetylcysteine, ascorbic acid and IV fluids was insufficient in this case due to a delay in initiation of treatment. The salient postmortem findings were icterus, subcutaneous and pulmonary edema and evidence of hemolysis in the liver, spleen and urinary tract.The pathophysiology of the toxicosis and the current recommendations for treatment are reviewed. PMID- 17422486 TI - Plasma cell pododermatitis in a cat. AB - Plasma cell pododermatitis, an uncommon disease of unknown etiology, is described in a six year old male domestic short-haired cat. The cat was referred with a history of lameness associated with swelling, softness and ulceration of the foot pads. The history suggested a seasonal occurrence of the condition. The dermis and subcutis of the foot pads were infiltrated by inflammatory cells which were mainly plasma cells. The large number of plasma cells present in the lesions suggests an immunological basis for the condition. PMID- 17422487 TI - Factors influencing time of parturition in range beef cattle. AB - A total of 2434 calving records from seven calving seasons were analyzed by the least-squares and chi-square procedures to study the influences of breeding group, age of dam, sex and birth weight of calf and dam weight at calving on the time of day of parturition. Frequency of day calving (0630 +/- 030 to 1900 h) was slightly but not significantly higher than night calving (1900 h to 630 +/- 030). The frequency of night calving was higher among heifers compared with the older cows. The cows which calved after May 15 (the last one third period of pregnancy) had higher frequency of day calving compared with those that calved earlier. The frequency of daytime calving was higher among the cows that weighed at most one standard deviation below the average compared to heavier cows of comparable age. PMID- 17422488 TI - A Brief Account of a Disease in Cattle Simulating Hemorrhagic Septicaemia due to Feeding Sweet Clover. PMID- 17422489 TI - The distribution of mycoplasmas and ureaplasmas in the genital tract of normal artificial insemination bulls. AB - Bull semen is commonly contaminated with mycoplasmas. To determine the source of contamination, semen and the genital tracts of 45 artificial insemination bulls were cultured for these organisms. The results indicate that mycoplasmas colonize the prepuce and the distal part of the urethra. Only rarely were they found in the ampullae or seminal vesicles. In 92% of the bulls with contaminated semen the same Mycoplasma species or Ureaplasma diversum was isolated from the prepuce and urethral orifice as was found in the semen. This suggests that the prepuce and distal urethra is the source of contamination. Colonization of the genital tracts with Mycoplasmas or U. diversum was not associated with histological changes. PMID- 17422490 TI - Herniation of small intestine through the right lateral ligament of the bladder in a bull. AB - A three year old Charolais bull was examined because of anorexia and depression of five days duration and progressive abdominal distention. Intestinal obstruction was diagnosed. A right flank celiotomy allowed diagnosis and correction of a herniated jejunoileal loop through the right lateral ligament of the bladder. An annular constriction located in mid-ileum was by-passed by ileocecal side-to-side anastomosis. The bull's appetite and fecal production returned to normal within 72 hours of surgery. Ten months following discharge the bull was reported to be well. PMID- 17422491 TI - Anestrus in Pigs: Confirmation by a Solid-phase RIA for Progesterone and Subsequent Response to Treatment. AB - All gilts not detected in estrus by eight months of age and any sows not observed in estrus by 14 days post-weaning, throughout a six month period in a University research herd, were bled on Monday and Thursday and plasma progesterone was measured by a solid-phase RIA. Basal concentrations of progesterone in both samples were considered indicative of true anestrus while one or more elevated values suggested that ovarian activity was present. Progesterone was basal in both samples from 44 of 70 gilts and all of the 23 sows tested. These anestrous animals were injected with either 400 IU PMSG plus 200 IU hCG or with 500 IU hCG plus 1 mg estradiol benzoate. The two treatments were equally effective with 60% of the injected females showing estrus and being successfully inseminated within ten days after treatment. Most of the gilts with elevated progesterone concentrations in the initial samples showed estrus within the following three weeks. The rapid solid-phase assay was able to differentiate between basal and luteal-phase levels of progesterone using unextracted plasma and could be a useful diagnostic aid for veterinarians attempting to determine major reasons for anestrus in gilts and sows. PMID- 17422492 TI - Problem solving in dairy health management. PMID- 17422494 TI - Oh no! Not another itchy dog? PMID- 17422493 TI - Veterinary medical diagnosis and disease control: clinical pathology. PMID- 17422495 TI - The modern role of meat inspection and food hygiene. PMID- 17422496 TI - Diagnostic procedures, prognosis and therapeutic approaches of chronic respiratory diseases in horses. PMID- 17422497 TI - Bovine brucellosis - diagnosis and eradication. PMID- 17422498 TI - Feed additives for the dairy herd: a literature review and discussion of practical applications. PMID- 17422499 TI - Recent advances in diagnosis of infectious diseases. PMID- 17422501 TI - Another field for the veterinarian. PMID- 17422500 TI - Si units in veterinary medicine - 1985. PMID- 17422502 TI - Testing for heartworm disease in dogs. PMID- 17422503 TI - Haemophilus somnus-induced Otitis and Meningitis in a Heifer. AB - The purpose of this report is to describe the lesions of otitis and meningitis associated with Haemophilus somnus infection in a heifer.The animal was found dead in its pen. At necropsy, there was a severe fibrinopurulent meningitis and suppurative otitis media and interna. The external ear canal contained inspissated debris. Haemophilus somnus was recovered in pure culture from the meningeal exudate. The identity of the organism was confirmed by fluorescent antibody and agar gel immunodiffusion tests. Possible pathogenetic mechanisms are discussed. PMID- 17422504 TI - Le choix d'un materiel de suture. AB - This review summarizes the physical properties of the most frequently used suture materials. It also describes the different healing processes involved in various tissues and discusses the main points of consideration in the proper selection of a suture material. PMID- 17422505 TI - Veterinarians in a changing world. PMID- 17422506 TI - Abdominocentesis in cattle: technique and criteria for diagnosis of peritonitis. AB - A reliable method for the collection of peritoneal fluid from cattle using a trocar and cannula is described. Peritoneal fluid was collected from three groups of cattle: periparturient, normal and with peritonitis. The fluid was examined by white cell count, differential cell count, total protein concentration and bacteriology. The results were analysed to determine the best criteria for peritonitis. Greater than 10% eosinophils were typical of normal peritoneal fluid. Peritoneal fluid with a relative neutrophil count greater than 40% and a relative eosinophil count of less than 10% was frequently associated with the diagnosis of peritonitis. Parturient cattle had large volumes of peritoneal fluid with low total protein and white cell counts. Growth of Gram-negative or anaerobic organisms was associated with mortality. PMID- 17422507 TI - Bat rabies in alberta 1979-1982. AB - The infection rate among eight species of bats submitted for rabies diagnosis in Alberta during 1979-82 was 4.6%. Prevalence of rabies was greatest (24%) for hoary bats Lasiurus cinereus, while the big brown bat Eptesicus fuscus was the species in which rabies was most commonly diagnosed, and the species submitted most frequently for rabies diagnosis was the little brown bat Myotis lucifugus. The rabies infection rate among male hoary bats was significantly greater than in either sex of all other submitted species. The frequency of rabies diagnosis in hoary bats submitted during 1979-82 was also significantly higher than in those submitted between 1971 and 1978. There has been a significant decrease in the rabies prevalence or infection rate of little brown bats since 1971-78. PMID- 17422508 TI - Evaluation of a Killed Vaccine Against Porcine Pleuropneumonia Due to Haemophilus pleuropneumoniae. AB - A bacterin containing serotypes 1 and 5 of Haemophilus pleuropneumoniae was developed for the prevention and the control of porcine pleuropneumonia. It was injected intramuscularly into three groups of ten piglets, the first group with one dose, the second one with two doses and the third one with three doses at two week intervals. Another group of ten piglets did not receive the vaccine. All the piglets were then challenged by an aerosol of mixed suspensions of H. pleuropneumoniae serotypes 1 and 5. Two and three injections of vaccine completely prevented mortality, whereas half of the control piglets and of those receiving only one dose of vaccine died. All surviving piglets, both control and vaccinated, had severe signs of respiratory disease for at least 36 hours after exposure to challenge. Moreover, vaccination did not induce the production of antibodies at high titers. Local reactions were not noted after vaccination and at postmortem; ten weeks after the challenge, there were no signs of abscess formation or induration. PMID- 17422509 TI - Bovine respiratory syncytial virus infection in an ontario cattle herd. AB - Bovine respiratory syncytial virus was recovered from the lung of a six month old calf that died during an outbreak of respiratory disease in a cattle herd in Ontario. Lung tissue removed from the calf at necropsy, performed within two hours of death, was frozen at -70 degrees C prior to virus isolation attempts. Syncytia and intracytoplasmic inclusions were demonstrated both in histological sections of the calf's lung and in stained cell culture preparations infected with the bovine respiratory syncytial virus isolate. Direct fluorescent antibody and virus neutralization tests serologically confirmed the identity of the isolate. PMID- 17422510 TI - Eradication of tuberculosis. PMID- 17422511 TI - Canine vaginal cytology during the estrous cycle. AB - Changes which occur in vaginal cytology of the bitch during proestrus, estrus. metestrus and anestrus are correlated. The terms superficial cell index and karyopyknotic index are introduced and explained in order that they may be used to describe vaginal cytological changes and to assist in breeding programs. PMID- 17422512 TI - Arteriovenous fistula in the orbit of a calf. AB - An Ayrshire calf with a protruding left eye and a fluctuant swelling dorsal to the orbit was examined. A diagnosis of arteriovenous fistula was made based on the findings of pulsation, a palpable thrill and a machinery-like bruit on auscultation over the swelling. Arteriography failed to outline the full extent of the fistula. The calf was euthanized and the fistula dissected. An enlarged superficial temporal artery communicated with a branch of the maxillary vein by way of a tortuous mass of vessels. The frontal bone was eroded by this plexus. PMID- 17422513 TI - An outbreak of diarrhea in piglets caused by a coronavirus antigenically distinct from transmissible gastroenteritis virus. AB - Coronavirus-like particles were visualized by electron microscopy in the intestinal contents of piglets during a diarrheal outbreak on a Quebec pig farm. The precipitating antigens of transmissible gastroenteritis virus were not detected in the intestinal contents of diarrheic animals by counter immunoelectrophoresis. Insignificant antibody titers against transmissible gastroenteritis virus were demonstrated in the sera of convalescent pigs by indirect immunofluorescence and these sera did not react with transmissible gastroenteritis virus when tested by immunoelectron microscopy. The causative agent could not be isolated in cell cultures. It was concluded that a coronavirus antigenically distinct from transmissible gastroenteritis virus was responsible for the enteric problems observed on this farm. The outbreak was controlled after oral inoculation of adult pigs with infected intestinal contents. PMID- 17422514 TI - Coccidiosis Caused by Isospora ohioensis-like Organisms in Three Dogs. AB - Three dogs with atypical coccidiosis are described. The age and clinical signs were not those normally associated with coccidiosis described in the literature. The ages of the three dogs were twenty-two months, five months and twelve weeks respectively. The clinical signs were recurrent mild to severe diarrhea, depressed growth and poor body condition. Repeated fecal examinations were needed to diagnose the disease. Sulfamethazine and amprolium were used successfully in treating the condition in all cases. PMID- 17422515 TI - An Economic Comparison of The Horse vs. Motor. PMID- 17422516 TI - Serum immunoglobulin levels in suckled beef calves: quantity or quality? PMID- 17422517 TI - The association between pneumonia and atrophic rhinitis in slaughter weight Swine. AB - Four hundred and sixty-two pigs from 37 herds were examined at slaughter for the presence and extent of turbinate atrophy and pneumonia. Turbinate atrophy was scored by measuring the mm space between the turbinate bone and the floor of the nasal cavity on both sides of the nasal septum. The total percentage of pneumonic involvement for each lung was evaluated by scoring the percentage of each lobe that was consolidated.There was a low, positive correlation between individual scores of turbinate atrophy and the associated percentage of lung involved with pneumonia (r = + 0.177; p < 0.001). There was a postive correlation between the herd mean turbinate atrophy score and the herd mean percentage pneumonia score (r = + 0.515; p = 0.001).The age at slaughter was known for 95 pigs from four herds and was not significantly correlated with the mm of turbinate atrophy, or the percentage of pneumonia. PMID- 17422518 TI - Plasma cell sarcoma in a cat. AB - Lytic lesions occurring in conjunction with plasma cell sarcoma (multiple myeloma) have rarely been reported in cats.A plasma cell sarcoma was diagnosed in a 13 year old castrated male Siamese cat with hind limb paresis resulting from osteolysis of the second lumbar vertebra. Serum electrophoresis showed a monoclonal gammopathy. A uniform population of plasma cells was found in and around the second lumbar vertebra and in the bone marrow of the femora, humeri, pelvis and the fifth lumbar vertebra. The neoplastic cells were identified as IgA and kappa chain specific by direct immunofluorescence. PMID- 17422519 TI - Concepts of disease, medical research and the challenges to the schools of the healing professions. AB - The study of disease is a significant part of the pattern of funding for medical research in North America and elsewhere. Also, the existence of disease and its importance in all branches of the healing professions is the major justification for separate professional schools of medicine. These considerations should encourage a vigorous exploration and development of concepts of disease as an important part of any medical education. Based on much of the current research activities, concepts of disease, especially chronic disease, seem largely outdated and not intimately reflecting the realization that the development of disease is often a physiological response to perturbations in the internal or external environment and not abnormal or pathological in the etymological sense. The importance of viewing cancer and other chronic diseases from this physiological point of view and not from the point of view of end-stage disease is emphasized by the use of one example, the development of cancer with chemicals. The challenge to the healing professions to develop more modern programs for educating the prospective research worker for the study of disease is discussed briefly. PMID- 17422520 TI - A new approach in the prevention of gastrointestinal parasitic infections in cattle. AB - The efficacy of a morantel long-acting device in preventing parasitic gastroenteric infections throughout a whole year was evaluated in heifers and steer calves in south central Quebec. Thirty-two calves, comprising nine Hereford steers, one Hereford heifer, fourteen Holstein crossbred steers and eight Holstein crossbred heifers, were allotted into two treatment groups and maintained throughout the grazing season on adjacent pastures resulting from equivalently dividing one original pasture. One morantel long-acting device was administered to each animal in one group at the time of turnout onto the pasture in the spring while the calves in the other group remained as nonmedicated controls. The parasitic infections incurred during the pasture season were primarily Ostertagia, Trichostrongylus, Cooperia and Nematodirus.The following fall, after twelve calves were necropsied for worm counts, the twenty remaining ones were brought into the barn where they were kept throughout the winter with access to an outside yard. They received good quality hay and rolled barley (1 kg/head/day) up until the following May, at which time they were weighed and had fecal samples taken for egg counts. In contrast to the results observed among controls, the morantel long-acting device treatment group had an 87% reduction in fecal worm egg excretion and a 67.3 kg per calf increased weight gain after one year. PMID- 17422521 TI - Use of oxytocin in sows. PMID- 17422522 TI - Cryptosporidiosis in two foals. AB - Cryptosporidium oocysts were identified by phase contrast microscopy on smears from flotations of greenish-yellow pasty feces obtained from two foals. One foal, a one week old Percheron was recumbent, anorectic and lethargic, believed to be the result of a septicemia of undetermined etiology. Despite therapy and nursing care the animal died. Using light and electron microscopy, numerous stages of Cryptosporidium sp. were seen protruding from the surface of epithelial cells of intestinal villi. The other foal, a six week old Arabian had a mild diarrhea. The diarrhea and passage of oocysts eventually ceased. Immunological tests on sera of both these foals provided no evidence of abnormal immune function. This report is the first to describe cryptosporidiosis in apparently immunocompetent horses. PMID- 17422523 TI - Submitting canine blood for prothrombin time and partial thromboplastin time determinations. AB - Practitioners commonly submit samples from dogs for partial thromboplastin time and prothrombin time determinations. Controversy exists as to the necessity for rapid separation of plasma and cells, and submission of the plasma on ice (or frozen). The purpose of this study was to address three questions. First, is it better to submit plasma or is whole blood satisfactory? Second, is it necessary to refrigerate the sample or is maintenance at room temperature (20 degrees C) adequate? Third, does the sample have to arrive at the laboratory within a few hours of collection or can reliable partial thromboplastin time/prothrombin time determinations be made on samples up to 48 hours old?It has been shown by this study that reliable partial thromboplastin time and prothrombin time determinations can be carried out on canine plasma for up to 48 hours after collection regardless of whether or not the plasma is separated immediately; however the samples must be kept at 4 degrees C. If the samples are maintained at room temperature, reliable prothrombin time determinations can be obtained for up to six hours after collection regardless of whether or not the plasma is separated immediately. Reliable partial thromboplastin time determinations can be made on plasma stored at 20 degrees C for up to 24 hours after collection and possibly longer (up to 48 hours) if the plasma has been separated immediately. PMID- 17422524 TI - Infection experimentale de porcs exempts d'organismes pathogenes specifiques avec une souche du virus de la grippe porcine (HSW(1)N(1)) et etude de la duree d'excretion virale. AB - EXPERIMENTAL INFECTION OF SPECIFIC PATHOGEN FREE PIGS WITH A SWINE INFLUENZA VIRUS (HSW1N1) AND STUDY OF THE DURATION OF VIRAL SHEDDING: A swine influenza virus (HSW(1)N(1)) isolated from an outbreak with respiratory disorders was experimentally inoculated to two groups of specific pathogen free pigs to study the pathogenicity of this strain and the period of shedding of the virus by the infected pigs. After the infection of the 12 oldest pigs (130 days old) no clinical sign was observed. Pneumonia was observed in three pigs of the 12 infected pigs slaughtered early after inoculation. The lung lesions were moderate and small. After the infection of the 22 seven week old pigs, hyperthermia appeared in 14 pigs and lasted approximately 48 hours. Lesions of pneumonia were seen on the lung of only one pig slaughtered eight days after the infection. The virus had been recovered from nasal swabs, on one pig, 29 days after the experimental infection, but all the contact trials with noninfected specific pathogen free pigs, 30-45 and 60 days after the experimental infection, failed. So, after experimental inoculation, the swine influenza virus weakly multiplied in pigs and its pathogenicity was moderate. In our experimental conditions, the shedding period of the virus was relatively short, less than four weeks. The possible reasons for the clinical reaction variation to the natural or experimental infection are discussed. PMID- 17422525 TI - Mammitis of the bovine. PMID- 17422526 TI - Borax toxicity. PMID- 17422527 TI - The development of the canadian veterinary profession. AB - A proposal for the development of Canadian veterinary education and of the organization of the profession is described. There should be one veterinary school with four branches (the current colleges). A student would train at any college in comparative medicine for two and one-half years and then train for 12 months or more in a specialty taught at one or more colleges. These specialties are general veterinary practice, poultry practice, public health and regulatory medicine, ruminant practice, swine practice, equine practice, small animal practice, fish medicine, fur-bearing and exotic animal medicine and research. After graduation in the chosen area there would be a period (six months or longer) of probationary licensing while field experience was gained before a final examination in the specialty was taken. The advantages and disadvantages of this proposal are discussed. PMID- 17422528 TI - Caprine beta-Mannosidosis in Kids from an Ontario Herd. AB - Caprine beta-mannosidosis, a fatal inherited deficiency of the lysosomal enzyme beta-mannosidase, was diagnosed in neonatal female Nubian crossbred twin kids from a small herd near Guelph, Ontario. The kids had been tetraplegic since birth, with whole body tremors, abnormal nystagmus and an intention tremor of the head.At necropsy, the histological lesions found consisted of widespread neuronal and visceral clear cytoplasmic vacuolation. Ultrastructurally, vacuoles were limited by a single membrane, and were empty or contained a small amount of amorphous dense material.Biochemical assay of sera and tissues confirmed negligible levels of beta-mannosidase activity, consistent with those of previously reported cases of caprine beta-mannosidosis.Vacuoles seen with light and electron microscopy are presumed to be lysosomes containing stored disaccharide and trisaccharide, the end products of incomplete catabolism of the oligosaccharide component of certain glycoproteins. PMID- 17422529 TI - Tetanus in a dog. AB - A three month old dog with tetanus was treated with antitoxin, antibiotic, a sedative and a muscle relaxant. A pharyngotomy tube was placed to facilitate feeding. The dog recovered after 23 days of therapy. PMID- 17422530 TI - Autumn abortions in sows. AB - This report describes the diagnosis and management of autumn abortions in a commercial swine herd in Ontario. A consistent pattern of abortions in the fall (and to a lesser extent, winter and spring) months was established from the herd records of 1980 to 1982. The percent abortions (of all farrowings) was 2.2, 3.7 and 4.3 for the years 1980, 1981 and 1982 respectively. Since laboratory examinations did not identify a specific pathogen, the management and the environment were investigated. In December 1982, supplemental heat was provided in the gestation barn to counteract heat losses at the level of the pregnant sows. The percent abortions was reduced to 0.4 over the next 12 months. PMID- 17422531 TI - Leptospirosis in sheep in Western Canada. AB - A survey of 930 ovine sera and kidneys from 33 sheep was conducted to assess the rate of leptospiral infection in sheep slaughtered in Alberta. Sera were tested for the presence of agglutinins to indigenous serovars of Leptospira interrogans. Kidneys with gross lesions were examined for the presence of leptospires by means of an indirect fluorescent antibody test (FAT) and by culture. Antibodies to serovars pomona and hardjo were present at rates of 1.0% and 0.4%, respectively, in sheep from Saskatchewan, Alberta and British Columbia. Sera from 120 feedlot lambs shipped from Oregon reacted to serovars pomona, hardjo and grippotyphosa at rates of 1.7%, 61.7% and 59.1%, respectively. Fluorescent antibody test detected serovars (presumptively) hardjo in 52% of Oregon feedlot lambs and grippotyphosa in 32% of the same group, a finding supported by the isolation of both these serovars from a pool of two fluorescent antibody test-positive kidneys. The grippotyphosa strain was highly virulent for hamsters, producing intense icterus and death. Leptospires, presumptively serovar grippotyphosa were demonstrated by fluorescent antibody test in one Alberta lamb kidney. The possibility of spreading leptospirosis by movement of breeding stock through public facilities and by assembling lambs in feedlots is discussed. PMID- 17422533 TI - My impressions of the meeting of the northwestern veterinary association. PMID- 17422532 TI - Growth Hormone-producing Pituitary Adenoma, Elevated Serum Somatomedin C Concentration and Diabetes Mellitus in a Cat. AB - A pituitary acidophil adenoma in a domestic shorthaired cat with diabetes mellitus and elevated serum somatomedin C level is described. Growth hormone production by the neoplasm was confirmed by an appropriate reaction using an immunoperoxidase technic. PMID- 17422534 TI - Monozygotic twins following micromanipulation of a frozen/thawed embryo. PMID- 17422535 TI - Reactions to a veterinary drug. PMID- 17422536 TI - Metabolic acidosis without clinical signs of dehydration in young calves. PMID- 17422537 TI - Dehorning with chem-cast. PMID- 17422539 TI - Clinical pathology in veterinary medicine. PMID- 17422538 TI - Radial-intermediate Carpal Bone Fusion in a Horse. AB - The radial carpal and intermediate carpal bones were found to be fused in a four year old Quarterhorse stallion having a subtle forelimb lameness. Clinical examination, diagnostic local anesthesia, radiography and arthroscopy were used to arrive at the diagnosis. A cause could not be established, although perinatal deformation of cartilagenous carpal bone templates is suspected. A similar case of carpal bone fusion was found among carpi collected for an anatomical study, indicating this condition may be more common than expected. Clinicians should carefully evaluate the flexed lateral view of the carpus for proximal movement of the intermediate and ulnar carpal bones in young horses presented for carpal lameness. The significance of this condition will remain uncertain until its incidence and relationship to lameness are better defined. PMID- 17422540 TI - Prevalence of Antibodies to Coxiella burnetii Among Veterinarians and Slaughterhouse Workers in Nova Scotia. AB - The complement fixation and the microimmunofluorescence tests were used to determine the prevalence of antibodies to Coxiella burnetii, the etiological agent of Q fever, among veterinarians and slaughterhouse workers in Nova Scotia. Seventeen percent of the 65 veterinarians and 12.5% of the 96 slaughterhouse workers tested had complement fixing antibodies to phase II C. burnetii antigen. Forty-nine percent of the veterinarians and 35% of the slaughterhouse workers had an antibody titer of >/= 1:8 to phase II C. burnetii antigen using the microimmunofluorescence test while 30% of the veterinarians and 14.5% of the slaughterhouse workers had antibodies detected to phase I antigen. Male veterinarians had a significantly higher rate of antibodies to C. burnetii phase II antigen compared with female veterinarians (p < 0.0087). An univariate analysis revealed that positive antibody titers (microimmunofluorescence test) to phase II antigen among veterinarians were significantly associated with exposure to cow, sheep and goat placentas; to stillborn calves, newborn foals, lambs and kids. By multivariate analysis the risk was highest for male veterinarians exposed to sheep placentas.Slaughtering cattle was a significant risk factor for positive antibody titers among slaughterhouse workers.We conclude that the high rate of antibodies to C. burnetii among Nova Scotia veterinarians and slaughterhouse workers is a reflection of the prevalence of Q fever in Nova Scotia and indicates that domestic ungulates are probably important in the epidemiology of Q fever in this province. PMID- 17422541 TI - Pemphigus and pemphigoid in domestic animals: an overview. AB - Pemphigus and pemphigoid are uncommon dermatological entities in domestic animals and of a presumed autoimmune nature. In one form or another, they have been reported in the dog, cat, horse and goat. Although these diseases are considered to be bullous dermatoses, the clinical presentation may vary from ulcerative to exfoliative to proliferative depending on the individual condition. Currently, four variants of pemphigus are recognized (vulgaris, vegetans, foliaceus, erythematosus) and two of pemphigoid (bullous, cicatricial) although cicatricial pemphigoid has not yet been conclusively demonstrated in animals. Diagnosis is based on history, clinical signs, histopathology and immunopathology. Therapy must be immunosuppressive to be effective and is palliative rather than curative. PMID- 17422542 TI - Prevalence of Antibodies to Swine Influenza Virus, Porcine Adenovirus Type 4 and Haemophilus pleuropneumoniae in Quebec Pig Farms with Respiratory Problems. AB - The prevalence of antibodies to three etiological agents involved in swine pneumonia was determined in Quebec pig farms which had experienced problems of mild to severe respiratory problems. Of the 350 sera collected from adults pigs, 67.0%, 18.3% and 46.6% had antibody titers to Haemophilus pleuropneumoniae type 1 and/or 5, to porcine adenovirus type 4 and to swine influenza virus, respectively. Comparatively, the serological prevalence rates for Haemophilus pleuropneumoniae, porcine adenovirus and swine influenza virus were 43.7%, 0% and 3.3% in farms not experiencing respiratory problems. PMID- 17422543 TI - Noninfectious nonerosive arthritis in two dogs. AB - Two cases of noninfectious, nonerosive arthritis are presented. The diagnostic features, clinical course and treatment are reviewed. Potential etiologies are discussed in the light of relatively little current material on this subject in the veterinary literature. PMID- 17422544 TI - Eradication of infectious bovine rhinotracheitis virus (bovine herpesvirus 1) from a herd of beef cattle. AB - Infectious bovine rhinotracheitis virus was eradicated from a 150 cow beef herd at the Animal Diseases Research Institute, Lethbridge, Alberta. Tests used to accomplish this included standard and modified serum-virus neutralization tests and an enzymelinked immunosorbent assay. These results and those of preliminary pilot studies in the herd and in a nonvaccinated, infectious bovine rhinotracheitis-infected 450 cow beef herd suggest that eradication of infectious bovine rhinotracheitis infection can be considered as a practical control alternative to vaccination, and that young animals in purebred herds could be monitored serologically and isolated, to enhance their eligibility for entry into artificial insemination studs or for export. PMID- 17422545 TI - Milk Fever: its possible association with derangements in the internal secretions. PMID- 17422546 TI - A Survey of Trichinella spiralis in Wild Carnivores in Southwestern Quebec. AB - In 1982-1983, 144 muscle samples from 11 different species of wild carnivores from southwestern Quebec, Canada, were examined for the presence of Trichinella spiralis larvae, using direct microscopic examination and the peptic digestion method. Two red foxes (Vulpes vulpes) (out of 29) and one pine marten (Martes americana) (out of 56) were found positive, giving a 2.1% prevalence for the whole sample. PMID- 17422547 TI - Pregnancy diagnosis in Swine: a comparison of two ultrasound instruments. AB - This investigation was designed to assess the sensitivity, specificity and positive predictive values of amplitude-depth and Doppler pregnancy detectors when utilized for pregnancy diagnosis of mated sows. Mated sows (n = 38) were tested daily from 15 to 45 days after mating for pregnancy with the ultrasound instruments. The same procedure was performed with nonmated sows (n = 10) 15 to 45 days after estrus. Both instruments were unreliable for pregnancy diagnosis between 15 and 22 days after mating. Between 23 and 45 days after mating the Doppler pregnancy detector was more specific and had greater positive predictive values. In contrast the amplitude-depth pregnancy detector was more sensitive during the same time interval.Serum estrone sulphate concentrations were determined in samples collected between 27 and 30 days after mating or estrus, as an alternate method of pregnancy diagnosis. Serum estrone sulphate concentrations were always equal to or greater than 0.5 ng/mL in the pregnant sows, while in the nonmated sows estrone sulphate concentrations were never more than 0.5 ng/mL serum. PMID- 17422548 TI - Education of the veterinarian. PMID- 17422549 TI - Endonucleases de restriction: isolement et identification d'une souche d'adenovirus canin de type 1. AB - For the first time in Quebec, a type 1 canine adenovirus was isolated in cell culture and typed by restriction endonuclease analysis. This virus originated from the internal organs of a young dog killed by a severe respiratory disease without showing any sign of hepatitis. PMID- 17422550 TI - Therapy of feline urethral obstruction. PMID- 17422551 TI - Stifle Lameness in Cattle at Two Veterinary Teaching Hospitals: A Retrospective Study of Forty-two Cases. AB - Records of two veterinary teaching hospitals from January 1, 1976 to June 1, 1982 were searched for diagnoses of stifle lameness. Forty-two records were found and information was recorded regarding signalment, history and clinical presentation. The following abnormalities were associated with stifle lameness: subchondral bone cyst (18 cases), joint instability (15 cases), degenerative joint disease (12 cases), cranial cruciate ligament injury (9 cases), cytological or bacteriological evidence of sepsis (9 cases), collateral ligament injury (3 cases), femorotibial luxation (2 cases) and intra-articular fracture (2 cases). The duration of lameness presentation ranged from 0.3 to 24 weeks and the mean follow-up period was 20.47 +/- 11.44 months (three animals were lost to follow up). Animals (n = 15) with subchondral bone cysts as the sole association with lameness presented at an early age (range - 6 to 18 months) and apparently regardless of treatment, had a good prognosis as determined by 75% (three lost to follow-up) returning to their intended function. Cattle (n = 9) with septic arthritis were presented at an age ranging from two months to seven years and only 22.2% returned to function. Cattle (n = 15) with joint instability presented at an age varying from nine months to 13 years also did poorly as only 26.6% returned to function. PMID- 17422552 TI - Canine vaginal cytology during the estrous cycle. PMID- 17422553 TI - Antibodies against equine herpesvirus 1 in the cerebrospinal fluid in the horse. AB - Neutralizing antibodies against equine herpesvirus 1 were measured in serum and cerebrospinal fluid of 16 horses and ponies from a closed herd both before and after vaccination with modified live equine herpesvirus 1. These titers were also measured in 22 neurologically normal and 15 neurologically abnormal horses at a teaching hospital. Animals from the closed herd had prevaccination serum titers up to 1:8 and postvaccination serum titers up to 1:128. Horses from the teaching hospital had serum titers up to 1:64. Cerebrospinal fluid titers were not detected in the vaccinated horses or the neurologically normal horses but a low titer (1:8) was noted in one neurologically abnormal horse. This titer probably resulted from hemorrhage into the cerebrospinal fluid following trauma. PMID- 17422554 TI - Sensitivity of routine tests for urine protein to hemoglobin. AB - Increasing concentrations of canine hemoglobin were added to aliquots of urine and saline to determine the relative sensitivity of several hemoglobin and protein detection methods including commercial reagent strips and sulfosalicylic acid. The hemoglobin detection pads of the reagent strips were 50 times more sensitive than the protein detection pads, indicating the presence of hemoglobin at a concentration of 0.001 g/L whereas the protein pads did not react positively unless the hemoglobin concentration exceeded 0.05 g/L. The sulfosalicylic acid test was the least sensitive, detecting hemoglobin only at concentrations of 0.4 g/L or higher. These results were similar for hemoglobin added either in the form of lysed red blood cells, intact red blood cells or associated with plasma proteins in whole blood.It was shown that a urine hemoglobin concentration eliciting less than the maximal score on the hemoglobin detection pad will not be detected as "protein" either with the commercial urinalysis strips or with sulfosalicylic acid. It was also seen that hemoglobin becomes visible as a red pigment when exceeding 0.3-0.5 g/L in a clear, light urine. It follows that a positive urine protein reading in the presence of a positive but less than maximal hemoglobin score or a protein reading exceeding 1.0 g/L in a nonpigmented urine indicates "true" proteinuria in excess of hemoglobin and plasma proteins associated with urinary tract hemorrhage. PMID- 17422555 TI - Milk Fever: its possible association with derangements in the internal secretions. PMID- 17422556 TI - Serological evidence of encephalomyocarditis virus in pigs in ontario. PMID- 17422557 TI - Diagnosis of TGE Virus Infection in Swine Herds. PMID- 17422559 TI - Leptospirosis in livestock. PMID- 17422558 TI - Embryo transfer in relation to bovine leukemia virus control and eradication. AB - Two hundred and seven, zona pellucida-intact bovine embryos were collected from bovine leukemia virus-infected donors, washed, and transferred to uninfected recipients: 111 of these embryos were sired by bovine leukemia virus-infected bulls. Fifty live calves were obtained from the 57 pregnancies resulting from the transfers. None of the recipients or calves developed antibodies to bovine leukemia virus. Nine zona-intact ova, 12 zona-intact morulae and 15 hatched blastocysts, exposed "in vitro" to bovine leukemia virus, washed and then tested for bovine leukemia virus were negative. Twenty-seven, zona-intact embryos and 14 hatched embryos were similarly exposed and washed prior to being transferred in groups to two uninfected recipients: no pregnancies resulted, nor did the recipients develop antibodies to bovine leukemia virus up to 120 days posttransfer. The conclusion from these and other bovine leukemia virus studies is that zona-intact embryos can be transferred from bovine leukemia virus infected donors, including those bred by bovine leukemia virus-infected bulls, without risk of transmitting bovine leukemia virus, providing that they are properly washed prior to transfer. PMID- 17422560 TI - Astrocytoma in a cat. AB - The presenting signs of a mature, spayed female, domestic shorthair cat with a thalamic astrocytoma were polyuria, polydipsia and behavioral changes of several months duration, followed by pacing and depression. Dexamethasone therapy temporarily improved the cat's mental status. A diagnosis of brain tumor was confirmed by postmortem examination. PMID- 17422561 TI - Fascioloides magna: Occurrence in Saskatchewan and Distribution in Canada. AB - Infection with Fascioloides magna, the large American liver fluke, was diagnosed in two moose (Alces alces) and six wapiti (Cervus elaphus) from central Saskatchewan. This is believed to be the first record of the parasite in the province. Fecal samples collected from wild wapiti at five sites in the commercial forest zone in Saskatchewan contained eggs believed to be those of F. magna. Trematode eggs were not found in feces from five captive herds of wapiti in the province, nor in samples from wild wapiti in Moose Mountain Provincial Park. Operculate eggs were found in feces of wild wapiti from Cypress Hills, but these were believed to be from trematodes other than F. magna. The distribution and significance of F. magna in Canada, based partially on responses to a mail questionnaire, are reviewed. PMID- 17422562 TI - Immune-mediated Hemolytic Anemia and Thrombocytopenia in the Dog: A retrospective study of 55 cases diagnosed from 1979 through 1983 at the Western College of Veterinary Medicine. AB - All recognized cases (n = 55) of immune-mediated hemolytic anemia and immune mediated thrombocytopenia in dogs presented to the Western College of Veterinary Medicine from 1969 through 1983 were reviewed. Specific areas of concern were: association with other conditions, therapeutic response, prognosis, relapse rate and final outcome. Of these 55 cases, 19 were immune-mediated hemolytic anemia, 26 were immune-mediated thrombocytopenia and 10 were both immune-mediated hemolytic anemia and thrombocytopenia. Females were slightly over-represented and the mean age was 6.4 years. Therapy consisted of various combinations of immuno suppressive drugs and in some cases, whole blood transfusion and splenectomy. No firm conclusions could be made regarding therapeutic efficacy, as a result of variation in treatment protocol and the occasional unavailability of follow-up data. Well over half of all cases were diagnosed as idiopathic. Precipitating factors or diseases most frequently implicated in secondary immune-mediated thrombocytopenia or hemolytic anemia were: recent vaccination, drug therapy, obstetrical complications, stress, recent viral infection and neoplasia. Twice as many cases of immune-mediated hemolytic anemia were seen in the cooler months (October to March), although this could not be related to antibody class or thermal reactivity. Immune-mediated thrombocytopenia both as a single disease and combined with immune-mediated hemolytic anemia had no seasonal incidence. History, clinical findings and hematological and clinical chemistry findings were consistent with data previously reported, with the exception of icterus, which appeared to be of higher incidence than most reports, being present in almost 50% of immune-mediated hemolytic anemia cases. Just over half of all dogs survived, although the survival rate was highest for immune-mediated hemolytic anemia, followed closely by immune-mediated thrombocytopenia and lowest for the combined disease. Immune-mediated thrombocytopenia most frequently ran a relapsing course requiring long-term or intermittent therapy. PMID- 17422563 TI - Antibiotic resistance associated with inositol metabolism in salmonellae from ontario cattle. AB - During 12 months in 1983-1984, Salmonellae were isolated from cattle 110 times. Salmonella typhimurium and Salmonella muenster were the most prevalent serotypes identified. All isolates were sensitive to polymyxin B. Isolates that did not produce acid from inositol were more frequently resistant to ampicillin, carbenicillin, kanamycin, sulfisoxazole, tetracycline and neomycin than isolates which could. Salmonella muenster isolates, which were almost exclusively inositol positive, demonstrated a high frequency of senstivity to commonly used antibiotics. The antibiotic sensitivities of the S. typhimurium isolates showed much greater variability, with multiple antibiotic resistance being frequently associated with those isolates that were inositol negative. PMID- 17422564 TI - Reminiscences of the great war - the aid extended by humane and kindred societies to the canadian army veterinary corps. PMID- 17422565 TI - Are pesticides involved in arthrogryposis? PMID- 17422566 TI - Poor Pay for AHTs. PMID- 17422567 TI - Canine vaginal cytology during the estrous cycle - a reply. PMID- 17422568 TI - Ocular absorption and toxicity of methotrexate in the dog. AB - The ocular absorption, elimination and toxicity of 12.5 mg of methotrexate in 0.5 cc administered subconjunctivally was studied in the dog. The purpose of this study was to establish ocular pharmacokinetics for treatment of intraocular tumours in dogs. Drug levels were measured in ocular compartments, and serum at 1,2,4,6,8 and 12 hours respectively. Tumoricidal concentrations of the drug were attained in the anterior chambers of the injected eyes up to and including six hours postinjection. Consistent signs of toxicity were anorexia, vomiting, weight loss and leukopenia. Local ocular toxicity following subconjunctival injection was minimal conjunctival hyperemia lasting up to 48 hours. PMID- 17422569 TI - Congestive heart failure associated with thymoma in two nubian goats. AB - Two mature Nubian does with signs of congestive heart failure were found at necropsy to have a large anterior mediastinal mass diagnosed histologically as thymoma. Lesions attributable to congestive heart failure were also present. PMID- 17422570 TI - Diagnosis of Leptospira serovar hardjo Infection in Cattle in Canada. AB - The investigations described were designed to identify the cause of serological reactions to Leptospira interrogans serovars hardjo and sejroe in Canadian cattle, and to confirm by culture a diagnosis of leptospirosis in cases of reproductive failure and atypical mastitis.Leptospires were detected in ten of 64 urine cultures, nine of 18 kidney cultures, and one of nine cerebrospinal fluid cultures. Twelve strains were purified. All were placed in the serogroup which contains serovars hardjo and sejroe. The nine strains which were fully serotyped were considered to be identical with serovar hardjo strain hardjoprajitno. Hardjo was isolated from cattle in the presence or absence of clinical disease and of antibody detectable by the microscopic agglutination test. Hardjo antigen was more sensitive than sejroe in detecting agglutinins in 58% of actively infected cattle and equal in 25%, as shown by comparative serum titrations. PMID- 17422571 TI - Postanesthetic poliomyelomalacia in a horse. AB - A clinically normal horse was anesthetized preparatory to surgery in dorsal recumbency for removal of a retained testicle. After recovery from the anesthetic, the horse was weak in the hind legs, subsequently deteriorated and became unable to rise and died on the eighth day after surgery. On microscopic examination, extensive poliomalacia of the thoracic and lumbar spinal cord was found. It is postulated that this lesion was a result of ischemic insult to the spinal cord during anesthesia and several possible pathogeneses are discussed. PMID- 17422572 TI - A study of foot disease of dairy cattle in quebec. AB - A 12 month survey of the different diseases of the digital region of dairy cattle was conducted in the province of Quebec in order to determine the relation between the frequency of the main digital lesions and different factors involved.Sole ulcer, white zone disease and erosion of the bulb represented 88.6% of all the lesions. The majority of the cases were recorded during the pasture season. An initial peak of prevalence was found in two year old heifers but cows were mainly affected between the ages of five to eight years. Sixty-six percent of the lesions appeared during the first three months of lactation. PMID- 17422573 TI - Occurrence of sulfa drug residues in canadian pork liver. AB - Sulfonamide residues in the livers of wholesome carcasses of Canadian pork were monitored over the six year period ending March 31, 1985. The annual rate of violative residues decreased from 9.92% to 2.75% over the course of the six years. At present the incidence of violation in the livers is 2.35% and there is no significant difference within the various regions of Canada. This would result in a muscle violative rate of 0.8%.The only sulfonamide found at violative levels was sulfamethazine. All violative samples were within the range 0.11-4.00 ppm. Over the period of the survey the incidence of violations at all levels decreased. Violations at higher levels decreased most rapidly resulting in a continuous reduction of the mean level in violative samples. PMID- 17422575 TI - Reminiscences of the Great War - The Aid Extended by Humane and Kindred Societies to the Canadian Army Veterinary Corps - Part II. PMID- 17422574 TI - Ovarian dysgerminoma in a bitch. AB - A nine year old Terrier-Chihuahua crossbred bitch was presented to the Western College of Veterinary Medicine with a history of anorexia, vomiting and diarrhea. A palpably enlarged uterus and purulent vaginal discharge were found on physical examination. Pyometra was diagnosed and ovariohysterectomy was subsequently performed. One ovary was twice the size of the other, firm and mottled grey-white on the cut surface. A dysgerminoma involving the larger ovary was diagnosed on histological examination. This tumor, closely resembling the male seminoma, was characterized by a uniform population of large round cells with scant acidophilic cytoplasm, frequent mitotic figures and focal aggregates of lymphocytes. History and clinical signs were considered to relate to the pyometra rather than the dysgerminoma. No conclusions could be made regarding hormonal elaboration by the tumor because sex hormone levels were not measured. Two years following discharge the dog was reported to be well. PMID- 17422576 TI - Prescriptions - medicated animal feeds. PMID- 17422577 TI - Epididymitis Caused by Brucella ovis in a Southern Ontario Sheep Flock. AB - Epididymitis was diagnosed in three rams in a commercial sheep flock in southern Ontario. The affected rams had palpably enlarged epididymides and two rams had semen which contained inflammatory cells and was of poor quality. Serum compliment fixation titers for Brucella ovis were 1:20, 1:80 and 1:90. Five other rams in the flock were clinically normal and without titers. Two of the affected rams had lesions similar to those produced by experimental infection with B. ovis. The infection in the rams had no apparent affect on ewe performance. The source of the infection remains unknown, but the rams were purchased from a flock which had imported ewes from the western U.S.A. PMID- 17422578 TI - Transitional Cell Carcinoma of the Urinary Bladder in a Beluga Whale (Delphinapterus leucas). AB - A transitional cell carcinoma of the urinary bladder was found in a beluga whale stranded in the St. Lawrence middle estuary. Various organs of this animal were submitted to high resolution gas chromatography coupled with mass spectrometry analysis. High frequency of urinary bladder cancer in the human population of the same area and the presence of carcinogenic compounds in the marine environment of this animal are discussed.Concurrent isolation of Edwardsiella tarda from various organs of this whale is also reported. PMID- 17422579 TI - Fanconi syndrome in a basenji. AB - Renal tubular dysfunction resembling canine Fanconi syndrome in a Basenji dog is described. The signs and laboratory results were similar to other reports of this disease. Therapy to correct some of the clinical signs and serum chemistry abnormalities was attempted. PMID- 17422580 TI - Facteurs influencant le profil metanologie des vaches laitieres. AB - Forty Holstein dairy cows were used to determine the effects of forage feeding system, season and stage of lactation on some blood parameters. Blood urea nitrogen was higher for cows consuming higher levels of crude protein and particularly for the ones on pasture. Hemoglobin and packed cell volume values were correlated with the level of crude protein intake and stage of lactation. Lower values of these parameters were observed with lower crude protein intake and in early lactation. Serum levels of albumin, calcium, inorganic phosphorus, sodium and potassium were not affected either by forage system, season or stage of lactation. Serum magnesium level was lower for forage system containing corn silage. Blood glucose was lower for cows in early lactation but was not affected by feeding systems during winter. Cows on pasture had higher blood glucose levels. Lower total protein concentration in blood was observed between 90 and 180 days of lactation as a result of lower gammaglobulins. Blood parameters are good indicators of nitrogen and energy nutritional status of dairy cows. However, blood mineral concentrations are not related to the level of mineral intake of animals. PMID- 17422581 TI - Comparison of common antibiotic therapies for haemophilus pleuropneumonia in pigs. AB - Three experiments were done to evaluate some antibiotic therapies that are used commonly to treat pigs infected with Haemophilus pleuropneumoniae. Haemophilus free piglets, 12 weeks of age, were challenged in a chamber with an aerosol of H. pleuropneumoniae serotype 1 and were medicated with antibiotics at various times before or after challenge. Antibiotic formulations which are commonly used to treat pneumonia in swine were used. They were chloramphenicol, penicillin, and a long-acting formulation of oxytetracycline given intramuscularly; and oxytetracycline, chloramphenicol and spiromycin (investigated as a potentially useful antibiotic) given in solution as the sole source of drinking water. Infection, disease (death, fever, gross lung lesions) and growth rate were measured in pigs following experimental challenge.The therapeutic effect of these antibiotic formulations was evaluated for prevention of the disease (52 pigs), treatment of acute disease (36 pigs), and treatment of chronic pneumonia (45 pigs). Injectable, long-acting oxytetracycline prevented all manifestations of disease (P<0.05) when given 24 hours before challenge. When treatment commenced immediately after the first signs of disease, each of the injected antibiotics reduced death rate (P<0.05), but they neither improved average daily gain nor reduced the incidence of infection and lung lesions. Chronically infected carrier pigs were produced by first immunizing them with a Haemophilus vaccine and then challenging them three weeks later. None of the treatments reduced the proportion of carriers of H. pleuropneumoniae. PMID- 17422582 TI - Induction of Parturition in the Cow using Cloprostenol and Dexamethasone in Combination. AB - Two experiments were designed to test the hypothesis that induction of parturition in the cow would be more predictable with the simultaneous use of a combination of cloprostenol and dexamethasone than with either hormone used alone.In experiment I all 19 beef cows treated with 500 mug cloprostenol and 25 mg dexamethasone in combination calved within 72 hours whereas dexamethasone (n = 19) or cloprostenol (n = 16) treatments alone each resulted in two induction failures. In those cows successfully induced, the mean interval from treatment to birth was 34.6 +/- 1.4 hours for the cloprostenol plus dexamethasone group, 43.3 +/- 2.4 hours for the dexamethasone group and 44.9 +/- 2.1 hours for the cloprostenol group. Control cows (n = 15) did not calve during the first 72 hours after treatment with saline. The incidence of retained placenta ranged from 19 to 53% in induced groups whereas placentae were not retained by cows in the control group.In experiment II all 30 beef cows in the cloprostenol plus dexamethasone group calved within the 72 hour limit, with a mean interval of 39.1 +/- 1.0 hours. Twenty-six of 31 cows calved within 72 hours with a mean interval of 51.9 +/- 3.4 hours after a single injection of cloprostenol and 29 of 33 cows calved within 72 hours with a mean interval of 52.6 +/- 3.3 hours after two injections of cloprostenol, 12 hours apart. Five of 34 control cows calved within 72 hours of time of treatment. The incidence of retained placenta was again high in induced cows. Results indicate that the simultaneous administration of cloprostenol and dexamethasone does constitute a safe, reliable and effective method of inducing parturition in the cow. PMID- 17422583 TI - Heartworm in dogs in Canada in 1984. AB - In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection. PMID- 17422584 TI - Leptospira interrogans serovar hardjo Infection in Cattle in the South Okanagan District of British Columbia. AB - An outbreak of leptospirosis due to Leptospira interrogans serovar hardjo in the South Okanagan District of British Columbia was investigated. The infection was associated primarily with bulls, but serovar hardjo was isolated from both bulls and cows at slaughter. Kidney and cerebrospinal fluid were found to contain leptospires, independently of the presence and level of serum agglutinins. Treatment of a bull twice in six months with dihydrostreptomycin failed to diminish an agglutinin titer (1/200) which persisted for two years without reexposure of the bull. A serological survey of cull cows sold through a central auction mart revealed the presence of hardjo agglutinins in 15.4% of 1300 sera representing 163 herds in 20 locations. Thirty percent of these herds contained reactor cattle. The number of premises from which reactor cattle came in a given locality varied from 4% to 67.7%. Measures to control leptospirosis in the study are suggested. PMID- 17422585 TI - Veterinary science in 1678. PMID- 17422586 TI - Upper respiratory obstruction in a thoroughbred foal. PMID- 17422587 TI - The influence of feedlot management on an outbreak of bovine respiratory disease. AB - Epidemic curves, odds ratios and chisquare were used to investigate an epidemic of respiratory disease in a pen of feedlot cattle. The cattle were divided into four groups by an eartag number received at processing. Data for each group were collected from feedlot records describing purchase, transportation and processing histories, daily feeding methods, daily pen movements, daily diagnoses, treatment and mortality rates. These data were used to describe the effects of market origin and feeding management on the levels and distribution of respiratory disease.The three groups of cattle purchased from auction markets and started on high levels of grain in their rations were determined to be 6.3 times (P<0.0005) more likely to be treated for any disease, 4.9 times (P<0.0005) more likely to be treated for respiratory disease, 12.7 times (P<0.0025) more likely to die, and 6.7 times (P<0.0471) more likely to die with respiratory disease than the group made up primarily of farm-assembled heifers and started on a 10% grain ration with time for adjustment to grain. PMID- 17422588 TI - Comparison of the bacterial and fungal flora in the pharynx of normal horses and horses affected with pharyngitis. AB - A total of 43 horses were used for the study of the pharyngeal bacterial flora. The median value of the number of bacteria in the group of 19 normal horses was 3.8 x 10(4) cfu/g of secretions. This value was 6.4 x 10(4)cfu/g in horses with grade I pharyngitis, 1.3 x 10(5) cfu/g in horses with grade II pharyngitis and 3.5 x 10(6) cfu/g in horses affected with grades III and IV pharyngitis. Corynebacterium spp, coagulase-negative staphylococci, Nocardia spp, Moraxella spp and Enterobacter spp were the most frequently encountered bacteria in the normal animals as well as in horses affected with pharyngitis of grades I or II. Moraxella spp were isolated in 87.5% of the horses with pharyngitis of grades III and IV, followed by Streptococcus zooepidemicus, Pseudomonas aeruginosa, coagulase-negative staphylococci and Enterobacter spp. No fastidious bacteria, nor strict anaerobes were isolated from any of the 43 horses. None of the microorganisms were found in 100% of the animals and the majority of the isolates were opportunistic bacteria. These results demonstrate that the isolation of Moraxella spp and S. zooepidemicus in large numbers is frequent in horses with lymphoid follicular hyperplasia grades III and IV. Fungi were isolated in small numbers from two or three horses in each group. PMID- 17422589 TI - Spontaneous External Endometriosis in a Gorilla (Gorilla gorilla). AB - The present report describes a case of external endometriosis in a 28 year old female gorilla (Gorilla gorilla). Microscopical examination of a pelvic mass observed at necropsy revealed ovarian tissue together with uterine glands and stroma, fibrous tissue and many siderophages. Theories of pathogenesis of external endometriosis are briefly reviewed. PMID- 17422590 TI - Effect of terbufos poisoning on the blood cholinesterase and hematological values in a dairy herd. AB - This report describes the effects of a sublethal dose of terbufos (counter 15G formulation) in a dairy herd. Ingestion of terbufos depressed whole blood cholinesterase activity. No recovery of this enzyme was evident until 30 days after ingestion after which a constant rate (0.65%/day) increase was observed.There was a good correlation between blood cholinesterase levels and clinical signs. Normal cholinesterase levels were reached at approximately 150 days after exposure. Cholinesterase levels in animals, ill prior to exposure, were slow to recover, while levels of healthy cattle surpassed those of control animals. Anemia was observed in some cows at approximately 30 days after exposure. The degree of anemia paralleled the severity of poisoning. The increase in packed cell volume paralleled the return of blood cholinesterase activity. PMID- 17422591 TI - Megaesophagus in a cat. AB - Megaesophagus in an eight month old Siamese cat is described. Initially, a cause for the vomiting was not discovered and the cat was treated for pyloric spasm. Several months later the same cat, in poor physical condition, was presented with a palpable bulge along its ventral neck. At this time a very dilated and flaccid esophagus was found. An exploratory thoracotomy was done but a cause for the megaesophagus was not discovered. PMID- 17422592 TI - Moldy sweet clover (dicoumarol) poisoning in Saskatchewan cattle. AB - Information pertaining to 286 animals from 56 herds affected by moldy sweet clover poisoning in Saskatchewan during 1983 was tabulated from the toxicology laboratory records. The morbidity in the affected herds was 12.1%, with a case fatality of 65.5%. Aborted fetuses and calves less than two weeks of age were affected with sweet clover poisoning most often. Sweet clover poisoning was more common during the period from January to April, 91.6% of the morbid animals were seen at this time. The rates of poisoning in beef and dairy cattle were estimated to be 0.156 and 0.095 cases per 1000 cattle respectively. Sweet clover poisoning was observed with the use of large of small bales of sweet clover in 78.9% of the affected herds. The geographical distribution of sweet clover poisoning in Saskatchewan followed the northern regions of the dark brown soil zone and the southern regions of the black soil zone which extend from the mid northwest portion of the province to the extreme southeast region.Dicoumarol concentrations were determined on a variety of tissues including liver, plasma, serum, kidney and muscle. Significant differences in the liver dicoumarol concentrations were found between animals of different ages (P = 0.045). Livers from younger animals, in particular, the fetus, contained lower concentrations of dicoumarol. PMID- 17422594 TI - The "compleat" jockey, 1688. PMID- 17422593 TI - Bovine stephanofilarial dermatitis in alberta. AB - The nematode, Stephanofilaria stilesi was recovered from two mature beef cattle in Alberta. The appearance of the skin and the histological lesions are described. The life cycle of the parasite and the development of lesions are reviewed. PMID- 17422595 TI - The Profession is Weakened. PMID- 17422596 TI - Canine distemper viral inclusions in blood cells of four vaccinated dogs. AB - Four cases of canine distemper were detected by the presence of numerous cytoplasmic inclusions in various circulating blood cells. Fluorescent antibody techniques and electron microscopy confirmed the identity of the viral inclusions. The cases occurred in the same geographic area and within a short time span. All four dogs had been vaccinated against canine distemper, but stress or other factors may have compromised their immune status. The possibility of an unusually virulent virus strain was also considered. PMID- 17422597 TI - A retrospective study of equine infectious anemia based on the canadian control program. AB - Equine infectious anemia in Canada was reviewed for the period January 1976 to December 1981. The human and ecological factors prevailing in Canada are deemed instrumental with respect to the evolution of the disease. The natural spread of the disease on a large scale has not been influenced by the Federal program. Reactors with signs of the disease are important for it's propagation. The author underlines the necessity of cooperation with private practising veterinarians to control it. PMID- 17422598 TI - Necrosis of the brain in calves following dehorning. AB - This is a report of an acute hemorhagic necrotizing encephalitis following dehorning in calves on pasture. Calves were found either dead or down in extremis. Multiple necrotic inflammatory lesions were found in the cerebral cortices of affected calves at postmortem. Necrosis and hemorhage were observed microscopically to be more prominent than inflammation, indicating the peracute overwhelming nature of the process. The etiology was not determined but clostridial infection was suspected. PMID- 17422599 TI - Comparison of methods for assessing the prevalence and extent of pneumonia in market weight Swine. AB - The purpose of this study was to compare four different techniques that may be used to assess the prevalence or severity of enzootic pneumonia in a swine herd. These techniques included: 1) assessing the percentage of lung involved and calculating a mean percentage and standard deviation for each herd, 2) counting the number of lungs in the herd sample that have greater than a predetermined amount of pneumonia and calculating the prevalence of the sample thus affected, 3) scoring only the lung from each herd sample that was maximally affected by pneumonia, and 4) allocating lungs to categories depending on the approximate extent of pneumonia. Five hundred and sixty pigs from 41 different swine herds in southern Minnesota were examined at slaughter and lungs were evaluated for the extent of pneumonia that was macroscopically visible.The mean percentage of pneumonia for a herd was positively and highly correlated with the standard deviation of that herd (r = +0.914; p < 0.0001), the prevalence of pigs having >/=5% lung involved with pneumonia in that herd (r = +0.946; p < 0.0001) and with the percentage of lung that was observed to be the maximally affected lung in each corresponding herd (r = +0.940; p < 0.001).The most informative procedure was assessing the percentage of lung involved and calculating a mean for the herd sample. Allocating lungs to categories based on approximate severity of pneumonia was less precise and more difficult to interpret. Assessing the prevalence of affected lungs, or evaluating the maximally affected lung was less time consuming and equally as informative for a herd-based indicator, as scoring the percentage of each lung and calculating a mean. PMID- 17422600 TI - A nutritional myopathy enzootic in a group of yearling beef cattle. AB - Steers and bulls under feedlot conditions and on an apparently adequate ration developed transitory signs of diarrhea and unthriftiness. One animal became recumbent. Sick and clinically normal animals in the group had elevated serum creatine kinase and aspartate aminotransferase enzyme values and degenerative changes in muscle fibers on biopsy. Feed analysis was carried out and the animals were monitored over several weeks by means of muscle biopsy and serum enzyme analysis as well as by postmortem examination at slaughter. Subclinical myopathy was found in several animals and was thought to be related to low vitamin E content in the high moisture corn ration. PMID- 17422601 TI - Physical and clinical pathological findings associated with experimentally induced rupture of the equine urinary bladder. AB - Two mature horses were examined for changes in laboratory and physical findings after experimentally induced bladder rupture. The postrupture laboratory diagnostic changes, which provide valuable information for a correct diagnosis are described. Hematology, serum and peritoneal fluid sodium, potassium, calcium, phosphorous, creatinine, urea nitrogen, albumin and peritoneal fluid components were measured and evaluated versus time. Hyponatremia and hyperkalemia occurred, as well as increased concentrations of peritoneal fluid potassium and inorganic phosphorus. In addition, peritoneal fluid creatinine:serum creatinine and peritoneal fluid urea nitrogen:serum urea nitrogen ratios were followed with time. Hematology and cytology of the peritoneal fluid showed an inflammatory response to urine contamination of the abdominal cavity. Physical findings of tachypnea and tachycardia as well as a mild colic were absent until nearly 50 hours postrupture. Based on these findings, it was concluded that the peritoneal fluid creatinine:serum creatinine ratio was the most useful antemortem laboratory diagnostic aid. PMID- 17422602 TI - Bovine mandibular fractures. AB - A retrospective study of bovine mandibular fractures was conducted. An increased incidence in males, beef breeds and animals less than one year of age was identified when the study group of 17 was compared to the total bovine case load. Manipulation during dystocia was the most common cause of fractures. Four neonates had rostral mandibular fractures. Fractures caudal to or involving the premolars were restricted to animals over a year of age.Treatment was attempted in ten of the seventeen cases, with euthanasia or slaughter elected in the remaining seven cases. Of the five cases treated by internal fixation, all four neonates died from conditions related to septicemia. PMID- 17422603 TI - Further extracts from "the experienced farrier". PMID- 17422604 TI - Shigella flexneri Type 2 Infection in Captive Nonhuman Primates. PMID- 17422605 TI - Alopecia areata in a dog. PMID- 17422606 TI - A giant step in disease eradication. PMID- 17422607 TI - Cerebellar disease in an adult cow. AB - This is the report of clinical signs and lesions of a cerebellar disorder in an adult four year old Limousin cow grazing perennial ryegrass (Lolium perenne). The most striking histopathological lesion was a marked paucity of Purkinje cells throughout the cerebellum. PMID- 17422608 TI - Calf-related Drug Use on Holstein Dairy Farms in Southwestern Ontario. AB - Calf-related drug use was studied on 104 randomly selected Holstein dairy farms in southwestern Ontario between October 1980 and July 1983. About 20% of dairy farmers were observed to administer antimicrobials preventively to healthy newborn calves (primarily as commercial "cocktails" of various antimicrobials, vitamins and minerals given in an oral bolus form) at some time during the year. About 30% of the farmers were observed to use vitamins (primarily vitamins A,D and E) preventively. The pattern of types of antimicrobials used preventively appeared to be different from the pattern seen for therapeutic use. Chloramphenicol was the single most commonly used therapeutic antimicrobial in this population of calves, accounting for some 30% of overall therapeutic antimicrobial use. In this study population, the prophylactic use of antimicrobials was associated with a decreased risk of being treated for pneumonia, and of dying, but an increased risk of being treated for scours. Case fatality rates for calves treated therapeutically varied, depending on the antimicrobial used and the routine of treatment. Calves treated with penicillin consistently suffered the highest case fatality rates. Case fatality rates for calves treated with chloramphenicol were considerably higher for calves treated once per day than for calves treated twice per day. For instance, calves first treated for scours with chloramphenicol, if treated once per day, suffered a case fatality rate of 22%. For scouring calves treated twice per day the case fatality rate was just under five percent. Clinical field trials of specific antimicrobials used preventively and therapeutically are required to delineate the conditions under which they might be useful. PMID- 17422609 TI - The effect of lindane formulations on tissue residues in barrows and on stillbirths in sows. AB - The potential effect of lindane and its formulations on stillbirths and abortion in pregnant sows was investigated. One of four formulations of lindane were applied at five times the registered dosage to each of ten sows within two weeks of farrowing. Each animal received 5 g of lindane. Formulations included: 1) a wettable powder diluted in water, 2) emulsifiable concentrate (EC) in xylene diluted with water, 3) an emulsifiable concentrate with heavy aromatic naptha diluted with mineral oil and 4) an emulsifiable concentrate ready-to-use mineral oil concentration. Number of stillbirths were not increased in the sows and signs of toxicity were not observed.Three barrows, for each formulation, were sprayed at three times the registered dosage (1.3 g/animal) and slaughtered 24 hours later to determine if increased absorption and residues of lindane were associated with different formulations. Residues of lindane in skin, fat, back meat, brain and liver were consistently higher in those animals sprayed with the water based formulations than with oil-based formulations. PMID- 17422610 TI - A Field Trial of a Pilated Moraxella bovis Bacterin for the Prevention of Infectious Bovine Keratoconjunctivitis. AB - Two field trials were conducted to evaluate the efficacy of a Moraxella bovis bacterin in the control of infectious bovine keratoconjunctivitis. The bacterin did not affect the incidence of infectious bovine keratoconjunctivitis in either trial but did appear to reduce the severity of lesions, treatments required and withdrawals from pasture required in one trial. Possible reasons for the apparently contradictory results are offered. It is suggested that the bacterin may be of some value when used in conjunction with other preventive measures. PMID- 17422611 TI - Laryngeal paralysis in a mature cat. PMID- 17422612 TI - Phalangeal and navicular bone hypoplasia and hoof malformation in the hind limbs of a foal. AB - Anatomical anomalies in the hind feet of a seven month old Appaloosa foal were identified and investigated through the use of gross anatomical dissection, radiography and angiography. Abnormalities were restricted to the distal aspect of both hind legs, the right hind leg being more severely affected. Anatomically the right foot resembled that of an equine fetus of approximately 120 days gestational age. Disruption of vascular perfusion to hoof structures was evident in both hind legs and was related to areas of abnormal bone conformation as well as to areas of abnormal ossification and calcification. Phalangeal and navicular bone hypoplasia were apparent as were soft tissue and joint anomalies. Although the etiology of the defects identified remains obscure, several theories are suggested, namely heritability, acquired defects and the possible teratogenic effects of clenbuterol. PMID- 17422613 TI - Euthanasia of reptiles - decapitation: an inhumane method of slaughter for the class "reptilia". PMID- 17422614 TI - Compendium of animal rabies vaccines marketed in Canada. PMID- 17422615 TI - PROCEEDINGS - Embryo Transfer in the Canadian Cattle Industry Status of Disease Transmission Studies and thier Relationship to the International Movement of Bovine Embryos. AB - The current, generally accepted approach to formulating health requirements for the international movement of embryos is to base them on the health status of the male and female donor animals. The alternative approach of basing them on the health status of the embryos themselves has been blocked by the lack of scientific information about the potential of the early embryo to transmit agents of infectious disease. Consequently, most research into infectious disease transmission by embryos has had the objective of assessing the potential of the embryo to transmit infectious disease, at the stage of development at which it is transferred commercially, with the thought in mind that, for some diseases, it may be possible in the future to focus on the embryo rather than the donor when drawing up health requirements for import permits. Results from experiments involving the bovine leukemia virus, bluetongue virus, infectious bovine rhinotracheitis virus, foot and mouth disease virus and Brucella abortus are encouraging to the point where, with the exception of foot and mouth disease virus, they could and should be put to the test in field studies. Research on several other bovine pathogens is underway, but the studies are not sufficiently advanced for a judgement to be made on the potential of embryos to transmit them. There is evidence that the research done is starting to have a positive effect through the relaxation of some health requirements for the international movement of embryos. PMID- 17422616 TI - The business of embryo transfer in Canada: opportunities and constraints. PMID- 17422617 TI - Current Embryo Transfer Technology: The Practitioner's View. PMID- 17422618 TI - Embryo Transfer in the Canadian Cattle Industry: Canada's Regulatory Approach. AB - In 1978, Canada was the first country to regulate animal embryo transfer in a manner similar to regulations for artificial insemination. Research has shown that the risk of disease transmission by bovine embryos is minimal therefore regulations are evolving to recognize this potential. The Canadian Embryo Transfer Association plays a key role in regulatory decisions. Importing embryos is based on risk assessment and on the impact of potentially introduced disease agents. Export strategy is based on negotiation of valid conditions, on effective certification of embryos and on full involvement at the international level. PMID- 17422619 TI - Artificial Insemination and Embryo Transplant: An Artificial Insemination Centre's Viewpoint. PMID- 17422620 TI - University of toronto, department of veterinary science, annual examinations, 1921. Fourth year. PMID- 17422621 TI - Development of a system for clinical evaluation of the biotin status of sows. AB - Sixteen test gilts were fed an egg white-fortified practical diet and four control gilts an egg white-free diet over a two-parity period in order to monitor changes in the serum biotin levels as induced biotin deficiency progressed. Gilts were individually housed in metal crates with slatted floors. Serum biotin (ng/L) of test animals declined from 1490 (initially) to 610 (month 3), remained stable (months 4-8) and thereafter approached 400, 135 and 30 after months 9, 13 and 15, respectively. Early clinical symptoms, i.e. lesions (on the feet, legs and skin), hoof cracks, hair loss and dry skin, were first observed after six months and became progressively more pronounced with time. After nine months, the induced symptoms resembled those characteristic of biotin deficiency. In contrast, serum biotin remained high in the control animals which did not develop symptoms. The data are interpreted to mean that serum biotin might be used as a sensitive index of biotin deficiency development in sows. On the basis of these observations, a system is proposed for evaluation of the sow's biotin status. Adequate biotin may be indicated by serum biotin level (ng/L) > 700; marginal, > 600 < 700; inadequate, > 400 < 600; and deficient, < 400. Alternatively, several concurrent clinical symptoms (i.e. alopecia, hoof cracks and lesions on the feet, legs and skin) may aid in the diagnosis of inadequate biotin status of sow herds reared under field conditions. PMID- 17422622 TI - La gestion sanitaire des elevages de rongeurs utilises en recherche biomedicale: I. La necessite de disposer d'animaux homogenes dans leur reaction. AB - THE NECESSITY FOR THE DISPOSAL OF HOMOGENEOUS REACTING ANIMALS: This paper points out the principal factors that may influence the course of experimentation using laboratory animals. Particular attention is given to extrinsic factors and to infectious agents. PMID- 17422623 TI - La gestion sanitaire des elevages de rongeurs utilises en recherche biomedicale: II. Les facteurs pouvant influencer I'implantation d'un programme de gestion sanitaire. AB - FACTORS INFLUENCING THE IMPLEMENTATION OF A MONITORING PROGRAM: This article outlines the principal factors that should be taken into account in a quality assurance program for rodents used in biomedical research. PMID- 17422624 TI - La gestion sanitaire des elevages de rongeurs utilises en recherche biomedicale: III. Le programme de surveillance des infections. AB - HEALTH SURVEILLANCE PROGRAM: This article outlines the principal components of quality control of laboratory rodents. PMID- 17422625 TI - Asceptic necrosis of bone in a dog. AB - Necrosis of intramedullary bone and bone marrow elements is reported in the long bones of a dog. Radiographically lesions were evident as an irregular increase in intramedullary density. On gross postmortem examination necrotic tissue appeared as yellow-white deposits on the endosteal surface and in the medullary cavity. Microscopically, necrotic bone had empty lacunae and was often covered by basophilic, woven bone. The cause and pathogenesisof aseptic necrosis of bone are discussed. PMID- 17422626 TI - Osteopetrosis in two foals. AB - This account describes two neonatal foals affected with osteopetrosis. One of these foals, a full term Peruvian Paso, was born alive, was bright and alert but was unable to stand and was euthanized at two days of age. The second foal, an Appaloosa, was also born alive at full term and was blind, weak, uncoordinated and unable to stand. Postmortem examination of both foals revealed generalized osteopetrosis and brachygnathia inferior. The osteopetrosis was characterized by failure of bone remodelling resulting in spicules of mineralized and nonmineralized cartilage covered by osteoid occupying the medullary areas of bones.It was not possible to determine whether this condition has an hereditary basis in horses. Circumstantial evidence suggested that the condition might be inherited in the Peruvian horse.The condition described in foals in this report and in one previous report is compared and contrasted with the disease in cattle. The lesions of osteopetrosis are similar in both species, but slightly less severe in horses. PMID- 17422627 TI - Leptospirosis: an occupational hazard to veterinarians. AB - Leptospirosis occurred in two veterinarians in Alberta, following their exposure to leptospires of domestic animal origin. The disease at onset resembled "flu" with fever, muscle and joint pain, and lassitude. It progressed through an extremely debilitating period with mild to severe hepatic and renal dysfunction, icterus and hemorrhage in one case, and cerebral meningitis in the other. Both patients were hospitalized for 11 to 14 days, where they responded to supportive and specific antibiotic and steroid therapy (penicillin G 10(6) IU q.i.d. and steroids, or tetracycline 500 mg q.i.d.). Diagnosis rested in one case on clinical signs and the observation of leptospires in blood and urine. In the other case, a tentative diagnosis of leptospirosis based on history and clinical signs was confirmed by serological test results and by the isolation of Leptospira interrogans serovar pomona from the patient's blood on day 6. Current occurrences of leptospirosis in man are reviewed. Convenient diagnostic methods, treatment and behavioural sequellae of leptospirosis are discussed. PMID- 17422628 TI - Ovotestes and sexual reversal in racing pigeons. AB - The occurrence of ovotestes associated with male behavioral characteristics in two mature female racing pigeons (Columba livia) is recorded. An ovotestis developed in the area of the vestigial right gonad of one bird and within the functional left ovary of the second bird. PMID- 17422629 TI - Recent advances in rabies diagnosis and research. PMID- 17422631 TI - Haemophilus somnus Complex: Pathogenesis of the Septicemic Thrombotic Meningoencephalitis. PMID- 17422630 TI - Haemophilus somnus: Bovine Reproductive and Respiratory Disease. PMID- 17422632 TI - Civil service of Canada - examination for positions of veterinary inspectors. PMID- 17422633 TI - Ascites in broiler chickens. PMID- 17422634 TI - Gutteral pouch mycosis and mycotic encephalitis in a horse. AB - A case of mycotic encephalitis in a horse with guttural pouch mycosis is described. A liquid pellet feed binder contaminated with Aspergillus sp. and erroneously mixed in a feed concentrate was a possible source of infection. PMID- 17422635 TI - Observations on intramammary infections in first calf heifers in early lactation. AB - Intramammary infections and mastitis were monitored on four occasions at three week intervals in 61 first calf heifers in five dairy herds during the first ten weeks of lactation. Of 940 quarter milk samples examined 65% were classed as negative, 10.4% as positive for mastitis (mainly subclinical), 1.8% as group 3 (infection present but no elevation in somatic cell count) and 22.8% as group 4 (elevated somatic cell count). Seventy-seven percent of the infections detected and identified were those due to coagulase-negative staphylococci, the main species being S. hyicus, S. epidermidis, S. simulans and S. hominis. Other infections detected with Corynebacterium pyogenes (three samples), Escherichia coli (one sample), Micrococcus spp. (one sample), S. aureus (two samples) and Streptococci (non-agalactiae) (seven samples).The geometric mean somatic cell count for 23 quarters infected with coagulase-negative staphylococci was 311 x 10(3) cells / mL compared to 134 x 10(3) cells / mL in noninfected adjacent contralateral quarters. The respective figures for% cell volume in Channel 8 (mainly neutrophils) were 10.6% and 3.5%. There was a highly significant association between herd and the proportion of quarter milk samples in the four mastitis categories. PMID- 17422636 TI - Porcine rabies in a closed feeder barn. AB - Rabies was diagnosed in a 40 kg pig from a closed feeder barn in which recently unexplained deaths of eight to ten pigs had occurred. A stray cat, probably crawling through a ventilation duct into the barn, was the suspected vector. Over a four and one-quarter year period a disproportionate 53 cases of animal rabies had been diagnosed within a 5 km radius of the farm (13% of the cases in the county in 3% of the county area). Histological lesions, mostly lymphocytic cuffing of blood vessels in the brain with lymphocytic meningitis, patchy to focal gliosis and satellitosis with neuronal necrosis were nonspecific and Negri bodies were rare. Fluorescent antibody technique examination of the brain for rabies virus antigen was positive. PMID- 17422637 TI - The existing and potential importance of brucellosis and tuberculosis in canadian wildlife: a review. AB - As the campaign to eradicate bovine brucellosis (Brucella abortus) and tuberculosis (Mycobacterium bovis) in Canadian livestock nears completion, the importance of extraneous sources of these diseases increases. This review summarizes the literature on brucellosis and tuberculosis in Canadian wildlife species to determine existing and potential hosts. Canadian caribou (Rangifer tarandus) are reservoirs of Brucella suis biotype 4 which is pathogenic in caribou, humans and muskoxen but reportedly nonpathogenic in livestock. Bison (Bison bison) and elk (Cervus canadensis) are significant reservoirs of B abortus and M. bovis. The bison in and around Wood Buffalo National Park have both diseases and are the only wildlife reservoir in Canada. Free-ranging elk are important reservoirs of brucellosis in Wyoming, and captive elk initiated the recent outbreak of bovine tuberculosis in 20 American states which has also involved bison and cattle herds. If bison and elk ranching continues to develop in Canada, the industry will have to be monitored to prevent the introduction and spread of infectious diseases like brucellosis and tuberculosis. This requires the evaluation and / or development of effective diagnostic methods for use in these animals. PMID- 17422638 TI - Suspected lead poisoning in an Amazon parrot. AB - A double yellow headed Amazon parrot (Amazona ochrocephala tresmariae) of unknown age and sex was examined for an acute onset of anorexia, listlessness, central nervous system signs and diarrhea. A tentative diagnosis of lead toxicosis was achieved based on radiographs, clinical pathology and response to therapy. Chelation therapy (Calcium EDTA) and supportive measures resulted in an uneventful recovery. PMID- 17422639 TI - Enquete eco-pathologique continue: facteurs de risque des mammites de la vache laitiere I. Analyses multidimensionnelles sur donnees d'elevage. AB - An epidemiological study about mastitis in French dairy herds, supported by an ecopathological survey is described.The aim of this work was to explain variations of the annual frequencies of clinical mastitis in the farms studied and of milk somatic cell counts as predictive of the infectious status of the mammary gland. Milk cell counts were expressed as annual percentages of cell results < 2.10(5) cells / mL (0.2 CEL) and > 1.10(6) cells / mL (1.0 CEL) in each herd.Twenty-nine herds were studied from June 1979 to July 1981 (22 to 92 cows per herd, Normande and Pie-Noire breeds) and, for the first time, the influence of 88 variables upon clinical mastitis, 0.2 CEL and 1.0 CEL was performed by multidimensional methods.These analyses led to the evidence of two kinds of factors linked with mastitis occurrences or milk cell-counts:1) Not well defined factors: geoclimatic, racial, links between production traits, extra-mammary pathology, susceptibility to mastitis and cells-counts; to elucidate if they were factors really linked with mammary infections, a complementary analysis is described as necessary (cf. second part of this study).2) Risk or protective factors, which appeared as: - absence of at least one annual examination of the milking machine (risk factor for clinical mastitis) - udder washing with individual towels, associated with enough straw for bedding (protective factor against mastitis) - teat dipping (associated with low milk cell-counts)Other prophylactic methods appeared without any effect. In the discussion, main difficulties of an epidemiological approach to the mastitis problem are summarized. A convenient hygiene-prophylaxis interaction is necessary to control mammary infections in dairy herds. PMID- 17422640 TI - Quality control performance in a provincial veterinary diagnostic microbiology laboratory system. AB - A quality control exercise was carried out in a provincial laboratory system to assess the performance of nine bacteriology technologists working in six laboratories. A group of ten mock specimens were distributed to each participant. Identification of all organisms and antibiotic susceptibility testing of pathogens was requested. Of the 117 organisms distributed, 104 were correctly identified, five were incorrectly identified and eight were not reported. Overall less than 1% very major errors, less than 5% major errors and less than 10% minor errors were observed in the antibiotic susceptibility testing. Most frequent errors were associated with penicillins, aminoglycosides, polymyxin, tetracycline and fastidious organisms. PMID- 17422641 TI - Foxes and fur bearing animals. PMID- 17422643 TI - Observations on a changing profession. PMID- 17422642 TI - Improved yersinia isolation from enteric specimens. PMID- 17422644 TI - The effect of increased mineral levels in the feed on leg weakness and sudden death syndrome in broiler chickens. AB - In two experiments with 98,000 and 40,000 broilers on commercial broiler farms, half of each flock were fed starter, grower and finisher diets with 0.2% added calcium, 0.2% added phosphorus and 0.2% added magnesium. This ration had no significant effect on the incidence of sudden death syndrome but the incidence of leg weakness in broilers on the increased mineral ration was higher in both flocks.Approximately 28% of the mortality in both flocks was from sudden death syndrome, more than from any other condition. Approximately 20% of the mortality was associated with leg weakness. PMID- 17422645 TI - The use of frozen semen from dogs in Canada. PMID- 17422646 TI - Rabies in Western Canada 1978-1984. AB - The results of laboratory examination of 22,503 specimens for the diagnosis of rabies by the standard fluorescent antibody and mouse inoculation tests over a seven year period are presented. Specimens were received from British Columbia, Alberta, Saskatchewan, the Yukon and Northwest Territories. Of the 1,445 positive cases, 10.50% involved human contact. The main reservoirs of rabies were skunks, bats and foxes. The reliability of the fluorescent antibody test for the diagnosis of rabies was reaffirmed by agreement with the mouse inoculation test in over 99% of cases. PMID- 17422647 TI - Giardiasis in a home. PMID- 17422648 TI - Lung sounds in cattle, horses, sheep and goats. AB - The purpose of this paper is to emphasize the importance of pulmonary auscultation for the clinician. It suggests a clarification and simplification of the terminology to be used which would be helpful to veterinary students and allow better communications between veterinarians. The interpretation of these sounds and the relationships to conditions and diseases of the lungs in cattle, horses, sheep and goats are discussed. PMID- 17422649 TI - Enquete eco-pathologique continue: facteurs de risque des mammites de la vache laitiere II. Analyses complementaires sur donnees individuelles et d'elevage. PMID- 17422650 TI - Prevalence of salmonella and campylobacter contamination in manitoba Swine carcasses. AB - Muscle samples from the neck and diaphragm and fecal samples were collected from five hogs from each of 50 lots of market hogs at three Manitoba abattoirs. Environmental samples were also collected. Each sample was tested for Salmonella contamination, and tissue and fecal samples were tested for Campylobacter.Salmonella was isolated from 34 (4.1%) of 821 samples, with 17 of the isolates coming from one lot of hogs and their concurrent environment. Campylobacter was isolated from 175 (26.3%) of 666 samples. Fifty-five percent were Campylobacter coli and 45% were Campylobacter jejuni. PMID- 17422652 TI - Poultry section - diseases of young chicks. PMID- 17422651 TI - Leptospirosis outbreak in a piggery in southern alberta. AB - Fourteen abortions, stillbirths and neonatal deaths occurred over an interval of one month in crated sows in a farrow-to-finish swine operation. Leptospira interrogans serovar pomona (kennewicki by DNA analysis) was demonstrated by culture and fluorescent antibody test to be present in sows, piglets, boars, feeders, drinking water and skunks on the premises.Antibody was found in all clinically affected sows, at serum dilutions ranging from 1/800 to 1/25,600, and in all breeding boars at titers from 1/50 to 1/1600. Pomona antibody was present in 118 sow sera collected nine months before the outbreak, at a prevalence of 21.3%. Parvovirus infection in fetuses was intercurrent with the leptospirosis epizootic, despite vaccination for the former. Environmental contamination, feedback through skunks via drinking water, and dissemination through the piggery are discussed. PMID- 17422653 TI - Braxy or Bradsot-like Abomastitis Caused by Clostridium septicum in a Calf. PMID- 17422655 TI - Anestrus and spontaneous galactorrhea in a hypothyroid bitch. AB - A bitch, with a history of prolonged postweaning galactorrhea, dermatitis and lethargy, was examined because of an extended anestrus and the development of spontaneous galactorrhea. A diagnosis of hypothyroidism was made based on the history, a subnormal T(4) level and the response to a therapeutic trial of thyroxine replacement. A possible pathogenesis for the spontaneous galactorrhea is discussed. PMID- 17422654 TI - Analogie entre les retrovirus humains lymphotropes et les retrovirus des grands animaux. AB - The family Retroviridae comprises some fifty viruses in three subfamilies: Oncoviridae, Lentiviridae and Spumaviridae. A better understanding of retroviral pathobiology has resulted from the rapid developments in knowledge of the molecular biology of normal and cancerous cells as well as retroviruses. Genomic relatedness was found between two human T cell leukemia viruses and bovine leukemia virus; similarly, some relatedness appears possible between human AIDS (acquired immunodeficiency syndrome) virus and lentiviruses of large animals. Because of their genomic relatedness, retroviruses from man and animals could theoretically form recombinants during in vitro manipulation. Therefore persons who work with retroviral materials should follow established laboratory practices to control infectious agents. PMID- 17422656 TI - Copper, zinc and manganese concentrations in equine liver, kidney and plasma. AB - Five groups of horses were fed different diets of known trace mineral concentration for a minimum of six months. Copper (Cu), zinc (Zn) and manganese (Mn) concentrations were measured in livers of 125 yearling horses and kidneys of 81 yearling horses as an assessment of trace mineral status. Plasma Cu and Zn determinations were made for all horses.Mean hepatic Cu concentrations of horses fed diets containing 6.9 to 15.2 mg Cu/kg dry matter (DM) feed were 0.27 to 0.33 mumol/g DM tissue. Plasma Cu concentrations ranged between 22.8 to 28.3 mumol/L. There was no simple mathematical relationship between plasma and hepatic Cu concentrations. Mean hepatic Zn concentrations in horses fed diets containing 25.6 to 52.2 mg Zn/kg DM feed were determined to be between 2.75 to 2.91 mumol/g DM tissue. Mean plasma Zn concentrations in groups of horses were between 11.7 to 13.5 mumol/L. Plasma Zn concentrations were not indicative of hepatic Zn concentration. Hepatic Mn concentrations ranged between 0.13 and 0.14 mumol/g DM tissue.Renal Zn concentrations ranged between 1.55 to 1.63 mumol/g DM tissue and did not differ with diet. Mean renal Mn concentrations were 0.09 mumol/g DM tissue for all groups of horses. Renal Cu concentrations ranged from 0.36 to 0.47 mumol/g DM tissue and differed with diet. PMID- 17422657 TI - Haemophilus somnus Mastitis in a Dairy Cow. AB - A clinical case of peracute bovine mastitis is described in which the most severely affected quarter yielded a heavy growth of Haemophilus somnus on culture. PMID- 17422658 TI - Corynebacterium equi Infections in Horses, 1958-1984: A Review of 131 Cases. AB - Of 131 cases of Corynebacterium equi infection in horses submitted for necropsy to the Ontario Veterinary College or Veterinary Laboratory Services, OMAF, Guelph, Ontario from 1958 to 1984, 115 were diagnosed as suppurative pneumonia, and of these 55 had associated ulcerative enterocolitis. Only five animals had intestinal involvement without pulmonary lesions. The remaining 11 cases included arthritis/cellulitis, skin abscesses and submandibular lymphadenitis. While the lung, intestine and associated lymph nodes yielded C. equi most frequently, in 21% of cases C. equi was also cultured from parenchymatous organs (spleen, liver or kidney) or blood. Corynebacterium equi infection accounted for 10% of all foals submitted for postmortem examination and 45% of all foals with pneumonia. Affected foals were one to four months of age. Submissions occurred between the months of May and August with a peak during July. There was a significantly greater prevalence of C. equi infection in Standardbreds when compared with other breeds. Of foals in this study, 36% were from farms which had had other horses succumb to this disease. Of the foals with pulmonary involvement, 21% did not have fever or clinical signs referable to the respiratory or gastrointestinal systems, findings which indicated that a large percentage of cases were subclinical. PMID- 17422659 TI - Accuracy of clinicians in predicting the bacterial cause of clinical bovine mastitis. AB - We examined the ability of clinicians to predict the causative organism of bovine mastitis in our practice. We obtained 118 milk culture results from 112 mastitic cows and compared the culture results to the predictions of clinicians at the time of milk sample collection. Sixty of 118 culture results were accurately predicted. The positive predictive value for coliform mastitis was 42% and the negative predictive value was 79% in a study population with a 31% prevalence of coliform mastitis. The proportion of coliform positive cultures accurately predicted was significantly greater than that expected by chance (p < 0.02). PMID- 17422660 TI - Du nouveau sur la maladie des griffes du chat. AB - AN UPDATE ON CAT SCRATCH DISEASE: Cat scratch disease is an uncommon and infectious disease of man characterized by regional lymphadenopathy following a scratch or a bite from a cat or other inoculation wounds. The role of the cat in the transmission of the disease is uncertain. This paper describes the disease, reviews different hypotheses on the etiology taking into account the recently reported discovery of a cat scratch disease bacillus and summarizes the diagnostic criteria. PMID- 17422661 TI - Horse section. PMID- 17422662 TI - Proposal for the regulation of canadian embryo services. PMID- 17422664 TI - International physicians for the prevention of nuclear war. PMID- 17422663 TI - The benefits of zootherapy. PMID- 17422665 TI - Rhinosporidiosis in a dog. AB - A dog with a nasal papilloma due to Rhinosporidium seeberi is presented. The causative organism was found using cytopathology and histopathology. The young Doberman dog had never travelled out of the province of Ontario and is the first case of rhinosporidiosis reported in Canada. PMID- 17422666 TI - Verminous Pneumonia in Adult Dairy Cows in Southern Ontario due to Dictyocaulus viviparus. AB - An outbreak of verminous pneumonia due to Dictyocaulus viviparus in a herd of mature, lactating, dairy cows in southern Ontario is described. The fact that the herd had been closed to new additions for ten years and had never experienced clinical disease due to D. viviparus in the past makes the occurrence of this herd problem difficult to explain. Correlation of fecal Baermann analysis for D. viviparus larvae with the progress of anthelmintic treatment is discussed. It is suggested that certain climatological variations in combination with unique, immunological aspects of D. viviparus infection may have contributed to the development of clinical disease in this herd. PMID- 17422667 TI - Les milieux de contraste en radiographie: proprietes, applications, indications et contre-indications, I. AB - The author describes the contrast media most commonly used in veterinary radiography. Emphasis is given to its physical properties and applications rather than examination techniques. There is also a reminder of the indications and contraindications for each examination using a given contrast medium. PMID- 17422668 TI - Bilateral carpus valgus with cranial bowing of the distal radius in a foal. AB - Bilateral carpus valgus with concomitant outward rotation and cranial bowing of the distal radii was diagnosed in a crossbred foal. The foal was not lame on admission and showed no radiographic evidence of carpal bone abnormalities. Surgery was limited to the most severely affected leg, and consisted of a combination of growth promotion (periosteal transection and stripping) and temporary physeal retardation (transphyseal bridging) procedures. Correction of the valgus deformity was nearly complete in the operated limb and substantial improvement was observed in the cranial bowing and outward rotation in both limbs, five months postoperatively. PMID- 17422669 TI - Ovine brucellosis in alberta. AB - Two parallel surveys of rams from Alberta sheep flocks were conducted to determine the presence of infection with Brucella ovis. In a retrospective study over a period of 24 months, using complement fixation test, 12 flocks out of 142 tested were considered infected. In another 17-month survey of slaughter rams by serology and culture methods 11 flocks out of 124 were found to be infected. The overall prevalence of ovine brucellosis was 8.6% of the flocks tested which represented 12.5% of the estimated sheep flocks in Alberta. Up to 67% of rams in infected flocks reacted to complement fixation test.The complement fixation test was evaluated for its efficiency in the diagnosis of ovine brucellosis and compared with a limited number of an enzyme-linked immunosorbent assay (ELISA) results and clinical criteria. The complement fixation test as well as ELISA identified all culture positive rams. Both serological tests appeared satisfactory for the diagnosis of B. ovis epididymitis when the results could be interpreted in the light of flock history and clinical findings. PMID- 17422670 TI - Dermoid sinus removal in a rhodesian ridgeback dog. AB - A three year old Rhodesian Ridgeback dog with a history of a small draining tract in the dorsal cervical area, was found to have a dermoid sinus on surgical exploration. The affected tissue was removed and histopathology done. The sinus is an inherited condition in this breed of dog, with surgical correction and neutering of affected individuals the treatments of choice to reduce the incidence of the dermoid sinus condition in the next generation. PMID- 17422671 TI - Transmission of Sarcoptes scabiei in Swine by Fomites. AB - Transmission of sarcoptic mange by fomites was investigated by placing mange-free piglets in pens for either fixed or variable periods of time during the first six days following removal of mange infected swine. Transmission occurred in pigs with as little as 24 hours exposure to fomites. Clinical signs of pruritis and focal erythematous skin lesions developed in various pigs from four and a half to 13 weeks after exposure. Pigs with the longer exposure developed clinical signs more rapidly than those with a shorter exposure. Four of six pigs developed a chronic form of the disease characterized by thickened encrustations and scurf from which mites were readily demonstrated. The remaining two pigs developed only the pruritic form and mites were never found in numerous skin scrapings examined. PMID- 17422672 TI - Cattle section. PMID- 17422673 TI - Buffalo - wood buffalo park. PMID- 17422674 TI - Multiple cutaneous mast cell tumors in a dog: a case report and brief review. AB - A spayed female yellow Labrador Retriever was presented to the small animal clinic at the Western College of Veterinary Medicine with an ulcerated, draining mass on the caudal aspect of the right thigh. Multiple small, hairless masses were also present on the thorax and in the flank folds. Fine-needle aspiration and cytological examination revealed well-differentiated mast cells and numerous eosinophils. A diagnosis of multiple cutaneous mast cell tumors was made. The clinical and cytological findings of this case are discussed, as well as the treatment and prognosis for mast cell tumors in general. PMID- 17422675 TI - Thyroid hormone levels in foals with congenital musculoskeletal lesions. AB - Fourteen foals with congenital or neonatal musculoskeletal abnormalities are described. Ten of the fourteen foals had abnormally low total serum T(3) and/or T(4) concentrations suggesting hypothyroidism. Response to thyroid-stimulating hormone was determined in two foals and found to be poor. Seven of the foals were necropsied and six of these had histological evidence of thyroid hyperplasia. These findings suggest hypothyroidism as a cause of congenital musculoskeletal lesions. PMID- 17422676 TI - Peripheral nerve sheath neoplasms in canadian slaughter cattle. AB - Two thousand four hundred and fifty-six bovine neoplasms were submitted during a ten year period of which 238 (9.7%) were neurofibromas. The neoplasms were mainly in the heart but also in the thorax, mediastinum and in some visceral organs. On gross examination 41 of the 99 cardiac neurofibromas were suspected to be Cysticercus bovis. Immunohistochemistry for S-100 protein and neuron specific enolase staining was used to confirm the diagnosis in several cases. PMID- 17422677 TI - The effect of azaperone on the agonistic behaviour of boars: a pilot study. AB - Previously unacquainted adult boars are often penned together and transported over long distances. This study examined the effect of azaperone, a drug used to reduce fighting in young pigs, on the behaviour of adult boars in close confinement.Five groups of six adult boars were used. Three groups were treated with 4% azaperone at 1.5 mg/kg, the other two groups served as controls. Behaviour was monitored for 24 hours. Azaperone reduced the intensity and ferocity of fighting during the peak activity of the drug but it did not eliminate aggressive behaviour. There was an increase in threat behaviour and the number (but not the intensity) of fights greater than one minute in the treated animals. This drug could be used when transporting boars in close confinement for short periods (less than four hours) if the boars are detusked. PMID- 17422678 TI - Dominion registration for veterinarians. PMID- 17422679 TI - Les milieux de contraste en radiographie: Proprietes, applications, indications et contre-indications, II. AB - The author describes the contrast media most commonly used in veterinary radiography. Emphasis is given to their physical properties and their applications rather than examination techniques. There is also a reminder of the indications and contra-indications for each examination using a given contrast medium. PMID- 17422680 TI - Treatment of Goats Infected with the Lungworm Muellerius capillaris. AB - Goats naturally infected with Muellerius capillaris were treated with ivermectin subcutaneously once or twice at the rate of 200 or 300 mug/kg body wt or with fenbendazole per os twice at 15 mug/kg body wt. Goats ceased passing larvae 11 to 20 days after treatment, and except for one doe, larvae reappeared in feces 34 to 59 days after treatment. In sections of lung of ivermectin-treated goats, adult Muellerius had swollen body walls and disrupted intestinal tracts. Granulomas, some mineralized, were present. It is suggested that immature Muellerius were not destroyed by either anthelmintic and that following destruction of the adults, immature Muellerius resumed development to the adult stage and produced more first-stage larvae. Treatment of Muellerius may be more effective if repeated after approximately a 35-day interval. PMID- 17422681 TI - Fistulography as an aid in the diagnosis of a nonradiopaque foreign body in a dog. AB - A case of chronic fistula associated with foreign body penetration of the soft tissues was diagnosed in the skull of a dog by fistulography. Radiographic technique and diagnostic features are described and illustrated. PMID- 17422682 TI - A system for the recording of clinical data as an aid in the diagnosis of bovine digital disease. AB - A form for the clinical evaluation of bovine digital disease was developed. In this article, each section of the resulting Digit Evaluation Form is discussed and justified.By following the Digit Evaluation Form guidelines in examination of 113 cattle (904 digits), objectives in two major areas were met: 1) Fundamental diagnostic techniques were consistently applied to clinical cases of bovine digital disease. 2) An accurate recording system was developed, largely due to the clinical application of a 12 zone system in graphic description of lesions. PMID- 17422683 TI - Sheep pox: experimental studies with a west african isolate. AB - Under conditions of a maximum security laboratory, four cross-bred sheep were inoculated intradermally only or intradermally and intratracheally with a West African isolate of sheep pox virus. All sheep had increased temperature and depression by the fourth or fifth day after infection. Nasal and lacrimal discharge and coughing occurred in all sheep but were more severe in sheep receiving the virus via the tracheal route. From the fifth day after infection, numerous papular erythematous skin lesions developed at the inoculation sites. These were 3-7 mm in diameter and gradually became nodular. Some of these lesions healed and others coalesced to form tumorlike masses. In one sheep, euthanized 14 days after intradermal and intratracheal inoculation, nodular lesions were found in the skin around the eyes, nostrils, oral and perianal regions, the mucosa of the rumen and throughout the lungs. Histologically, skin nodules were characterized by ischemic necrosis, vasculitis, microvesicualtion, eosinophilic cytoplasmic inclusions in the dermal epithelial cells and vacuolar nuclear degeneration. The pulmonary lesion was that of proliferative alveolitis with occasional cytoplasmic inclusions in the alveolar cells and macrophages. Ultrastructurally, large cuboidal virus particles were found both in the skin lesion and inoculated tissue cultures. The sheep pox virus structure was easily distinguished from contagious ecthyma virus, a parapoxvirus which causes sporadic disease in Canada. Serum neutralizing antibodies developed in all the sheep by 14 days postinfection.The clinical and pathological characteristics of experimental sheep pox produced with this West African isolate were similar to those caused by Neethling virus of lumpy skin disease in cattle. PMID- 17422684 TI - Management and housing factors associated with piglet preweaning mortality. AB - The purposes of this study were to establish piglet preweaning mortality rates on representative Ontario pig farms and to examine possible relationships between preweaning mortality and various management and environmental factors. Thirty randomly sampled farrow-to-finish herds were surveyed over a two year period. Mean preweaning mortality in liveborn pigs of 18.5+/-5.8 and 18.7+/-5.4 was recorded in the first and second years of the survey, respectively. Preweaning mortality was correlated positively with still-birth rate (r(1)=0.31, p(1)<0.1 (Year 1), and (r(2)=0.37, p(2)<0.05 (Year 2)) and with weaner pig mortality in the second year (r(2)=0.37, p(2)<0.05). Preweaning mortality was negatively correlated with herd size (r(1)=-0.30, p(1)<0.1 and (r(2)=-0.31, p(2)<0.1), crate utilization (r(1)=-0.52, p(1)<0.01 and (r(2)=-0.53, p(2)<0.01), and positively correlated with the number of farrowing crates per room (r(1) and r(2)=0.52, p<0.05).There was no relationship observed between preweaning mortality and housing and management characteristics such as the use of bedding or no bedding, batch farrowing versus continuous farrowing and single-use compared to multi-use farrowing rooms.It was felt that stockmanship was an important modifier of environmental effects. PMID- 17422685 TI - Elephants in war. PMID- 17422686 TI - Letters to the editor. PMID- 17422687 TI - Letters to the editor. PMID- 17422688 TI - Canine Von Willebrand's Disease: A Common Inherited Bleeding Disorder in Doberman Pinscher Dogs. AB - Von Willebrand's disease is the most common inherited bleeding disorder of dogs occurring with particularly high frequency in Doberman pinscher dogs. Because of its method of transmission (autosomal incomplete dominant), the clinical and laboratory severity of the disease varies considerably. "Stress" may be required to make the increased bleeding tendency clinically apparent. This report describes five cases of Von Willebrand's disease in Doberman pinscher dogs and illustrates the variety of clinical expressions that the disease may take. PMID- 17422689 TI - Leiomyoma of the uterus of a mare. AB - A mare was presented for diagnosis and treatment of infertility of three seasons duration. Examination revealed a uterine tumor. The tumor was removed surgically and diagnosed histopathologically as a leiomyoma. The mare conceived and successfully foaled after the surgery. PMID- 17422690 TI - Distempter vaccination of dogs: factors which could cause vaccine failure. AB - Distemper vaccination failures are uncommon. A number of factors which could cause such failure are discussed. The blocking effect of maternal antibody can be expected in 50% of pups at six weeks but is not important after 12 weeks. Among intercurrent infections, the immunosuppressive effect of parvovirus has the potential to precipitate vaccine-induced distemper. Corticosteroids at levels up to 10 mg/kg do not interfere with successful distemper vaccination. Anesthesia or surgery has little effect but use of chloramphenicol or tetracyclines should be avoided. High environmental temperatures can lead to vaccine failure, as can vitamin E deficiency. Excessive environmental exposure to virulent distemper virus can overcome levels of protection which would be adequate under normal circumstances. PMID- 17422691 TI - Heartworm in dogs in Canada in 1985. AB - In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada. PMID- 17422692 TI - The Persistent Isolation of Salmonella typhimurium from the Mammary Gland of a Dairy Cow. AB - The contamination of cheese by Salmonella typhimurium serotype 10 resulted in the bacteriological examination of raw milk from 327 farms on Prince Edward Island. A milk sample from a bulk tank from one farm was positive for this pathogen. The possible source was linked to the isolation of the strain from the milk from one quarter of one cow. Although the cow was asymptomatic for any clinical illness other than a chronic staphylococcal mastitis in two quarters, the animal continued to shed Salmonella in the milk during a 36 day period. Necropsy failed to reveal a source of the infection. PMID- 17422693 TI - Episodic muscle tremors in a quarter horse: resemblance to hyperkalemic periodic paralysis. AB - A three year old Quarter Horse stallion was presented with a one year history of episodes of generalized muscle tremors and stiffness, and spasm of the muscles of facial expression, lasting 10-15 minutes. Between attacks, the horse was either normal or had a localized muscle tremor in the flank region. Episodes appeared unrelated to exercise. The major abnormal findings included 1) a rise in plasma potassium from a resting level of 4.4 to 7.9 mmol/L during an attack and 2) electromyographic findings of generalized increased insertion activity and myotonic discharges. The horse was treated with hydrochlorothiazide tablets for nine months, during which time no further attacks were noted. However, four months after the drug was stopped, sporadic focal muscle tremors reappeared; two months later, generalized attacks were seen. Despite reinstitution of the diuretic, a focal flank tremor persisted. Two related horses in the same stable also were reported by the owner to exhibit sporadic generalized muscle twitching. The abnormal findings of the present case differ from clinical syndromes previously reported in horses. Some similarities to hyperkalemic periodic paralysis in humans are noted. PMID- 17422694 TI - Letter to the editor. PMID- 17422695 TI - Historical Column//Page d'histoire: GRIFFITH EVANS 1835-1935 DISCOVERER OF THE FIRST PATHOGENIC TRYPANOSOME. PMID- 17422696 TI - Together in '87! PMID- 17422697 TI - Animal rights, human rights? Veterinarians as prisoners of conscience. PMID- 17422698 TI - Profile. PMID- 17422699 TI - Borzoi pups from frozen semen. PMID- 17422701 TI - Role of BVD in Bovine Abortion. PMID- 17422700 TI - On abortions. PMID- 17422702 TI - Ivas. PMID- 17422703 TI - Cross-Canada disease report. PMID- 17422704 TI - Protein energy malnutrition and fat mobilization in neonatal calves. AB - Fat stores and organ weights were assessed in calves at birth (n=5) and after seven days of milk (n=5) or electrolyte (n=5) feeding.Compared to newborn calves, milk-fed calves had a significant (p < 0.05) redistribution of fat from perirenal area to bone marrow. The thymus also involuted during milk feeding.In electrolyte fed calves there was a significant loss of perirenal and bone marrow fat. The visible omental, mesenteric and subcutaneous fat stores were depleted. Epicardial fat stores were not visibly affected.There was a high correlation between bone marrow crude fat and bone marrow dry matter (R=0.92). This suggests that dry matter estimations can be used to assess bone marrow fat stores. Perirenal fat may be intermediate in type between brown and white adipose tissue because it is mobilized in response to fasting, and formalin fixed perirenal fat did not contain detectable levels of thermogenin. PMID- 17422705 TI - Canine blastomycosis in southern Saskatchewan. AB - The incidence of canine blastomycosis in southern Saskatchewan is examined and three clinical cases are described. Nineteen cases of the disease have been diagnosed in southern Saskatchewan since April of 1981. Eight cases were diagnosed during a six month period from August 1985 to February 1986 in dogs residing in a small central area of Regina. The geographical and chronological clustering of cases suggests a local source of exposure to Blastomyces dermatitidis, not previously considered to be endemic to Saskatchewan. PMID- 17422706 TI - The knobbed acrosome defect in beef bulls. AB - The knobbed acrosome defect was found at levels of 25 to 100 percent of spermatozoa from 16 of 2054 beef bulls. The incidence of this defect appeared to be particularly high in the Charolais breed. Pedigree analysis of some of the affected Charolais bulls indicated there may be a genetic predisposition for this sperm defect. In eosin-nigrosin stained semen smears the most common form of the abnormality was a flattened or indented apex of the sperm head. A refractile bead at the apex of the sperm head was seen less commonly. Electron microscopy of the spermatozoa from one bull showed that the abnormality was similar to the knobbed sperm defect previously described in Friesian bulls. A breeding trial confirmed that bulls producing spermatozoa with a high incidence of knobbed acrosomes are infertile. PMID- 17422707 TI - Successive torsion of the right middle and left cranial lung lobes in a dog. AB - This case report describes the torsion of two lung lobes in a dog. The animal was first presented for a torsion of the right middle lung lobe. Following the surgical resection of that lobe, the dog suffered another torsion of the left cranial lung lobe (cranial and caudal segments). PMID- 17422708 TI - Monensis toxicity in lambs. AB - A group of 100 purebred lambs was accidentally fed ten times the prescribed level of monensin in its grower ration. The first clinical sign was feed refusal. When the lambs were forced to eat the ration by withdrawal of other feeds, clinical signs developed within 48 hours in 15 to 20% of the group. These included recumbency, inability to rise, stiff gaits and a tendency to walk on their toes. Clinical examination, history, laboratory and necropsy results, along with feed analysis confirmed the diagnosis of monensin toxicosis. PMID- 17422709 TI - Rinderpest and mainstream infectious disease concepts in the eighteenth century. PMID- 17422710 TI - Ivadm. PMID- 17422712 TI - Cross-Canada disease report. PMID- 17422711 TI - Leptospirosis in farm animals: the hidden disease. PMID- 17422713 TI - Canadian holsteins in nicaragua. PMID- 17422714 TI - Omission. PMID- 17422715 TI - Intranuclear inclusion bodies in finches. PMID- 17422716 TI - The Successful Management of Leptospirosa hardjo Infection in a Beef Herd in Northern Ontario. AB - Abortion, premature calving, hemolytic anemia and fatal hematuria were associated with high levels (titer > 10(-4)) of antibody to Leptospira interrogans serovar hardjo and with isolation of hardjo in a herd of 265 beef cattle in the Great Clay Belt of northern Ontario. This herd was bred by artificial insemination, after heat detection by vasectomized bulls. The antibody prevalence rate in the herd was 54 to 60% over a five year period. The rate tended to reach 100% by age three years and to be below 5% in yearlings, which were raised in isolation from older cattle. Hardjo was isolated from the urine of a cow that aborted in the eighth month of pregnancy, and from kidneys of yearling steers which had been exposed to an older cow. Maternal antibody levels in calves paralleled those in their dams, protecting calves while they were being naturally exposed to infection, thus contributing to the achievement of balance between host and parasite. A controlled vaccination trial was conducted in 50 initially seronegative yearling steers and heifers. Serological response to vaccine was limited to a maximum agglutinin titer of 10(-2) in 8% of vaccinated cattle. Vaccination reduced the infection rate from 86% in the controls to 46% in the treated group, indirectly reducing the number of calves for which colostral antibody against hardjo would be available. A vaccination program was not implemented in the herd. Hardjo infection appeared to die out over a period of six years following the initial five year study period, with antibody prevalence falling from 60% to 0.7% and reactors persisting only in two eight year old cows. Decline in infection was coincident with changes in management which protected heifers from exposure to infection until their third pregnancy, and which probably lowered the reservoir of infection by increased culling from older age classes. PMID- 17422717 TI - Leptospirosa pomona Abortion Storm in a Cattle Herd in Saskatchewan. AB - Abortions occurred in 18% of 131 beef cows and heifers during two months, on a farm in southern Saskatchewan. The losses began two weeks after acute febrile illness and agalactia in a dairy cow to which the beef herd had been exposed. A diagnosis of Leptospira interrogans serovar pomona infection was made on the basis of serology in cows and the finding of leptospires in fetal tissues by fluorescent antibody test. Tentative diagnosis of infectious bovine rhinotracheitis delayed treatment and prophylaxis until infection attained high intensity in the herd and severe losses to the farmer occurred. Abortions ceased after vaccination against pomona and oxytetracycline treatment of pregnant cows, although chronic debility followed the acute phase of the disease in some cows. Recrudescence of infection was suspected four months later, when acute agalactia occurred in one cow and debility in calves and cows was recurring. Pomona infection was not proven, but dihydrostreptomycin treatment and revaccination were applied to the whole herd. Seroconversion and IgM antibody continued to indicate a persistent source of infection and susceptibility in a minority of the population one year after onset. The source of the original infection is believed to have been a carrier beef cow, or a dairy cow which was leptospiruric at the time of contact with the beef herd. With the exception of one aborted calf, no evidence of pomona infection was found outside the farm, in cattle or wild mammals tested serologically within a radius of 30 km, during one year following the outbreak. PMID- 17422718 TI - Anticoagulant-induced Alterations in Pig Platelets. AB - Application of an anticoagulant to prevent clotting is a prerequisite for the retention of morphological integrity of platelets in blood samples. Among the anticoagulants used for the collection of blood from veterinary animals, are heparin, the oxalate salts (sodium and potassium) and di-sodium and tri-sodium citrates. In a study on the effects of anticoagulants on platelet morphology, it was noted that oxalates caused severe morphological distortion in platelets. It is hypothesized that the membranous extensions, pseudopodia and loops detected in the platelets in oxalated blood may have been triggered by the irreversible formation of calcium oxalate salts which in turn alter the intracellular electrolyte balance. PMID- 17422720 TI - Rinderpest and Mainstream Infectious Disease Concepts in the Eighteenth Century: Part II. PMID- 17422719 TI - Analogy between lymphotropic human retroviruses and large animal retroviruses. AB - The family Retroviridae comprises some fifty viruses in three subfamilies: Oncoviridae, Lentiviridae and Spumaviridae. A better understanding of retroviral pathobiology has resulted from the rapid developments in knowledge of the molecular biology of normal and cancerous cells as well as retroviruses. Genomic relatedness was found between two human T cell leukemia viruses and bovine leukemia virus, similarly, some relatedness appears possible between human AIDS (acquired immunodeficiency syndrome) virus and lentiviruses of large animals. Because of their genomic relatedness, retroviruses from man and animals could theoretically form recombinants during in vitro manipulation. Therefore persons who work with retroviral materials should follow established laboratory practices to control infectious agents. PMID- 17422721 TI - A survey of ectoparasites on cattle in central alberta, november 1984-july 1985. PMID- 17422722 TI - 1. The evolution of computer technology in veterinary medicine. PMID- 17422724 TI - Dr. Richard Pattie...In his own words. PMID- 17422723 TI - The reindeer industry in Canada. PMID- 17422725 TI - Cross-Canada disease report. PMID- 17422726 TI - A review of morbidity and mortality rates and disease occurrence in north american feedlot cattle. AB - A review of veterinary literature on morbidity or mortality rates in feedlot cattle was performed. Incidence (attack) rates were the only types of rates reviewed. Differences in the definition of terms made reports difficult to compare. Case-definitions were often poorly defined and most were based on chemotherapeutic treatment as a criterion. A summary was made of 14 comparable studies containing disease incidence rates in calves in the first few weeks following arrival in feedlots. The incidence of morbidity ranged from 0% to 69% with most reports between 15% and 45%. The mortality rate in the same period ranged from 0% to 15% with most reports between 1% and 5%. The peak incidence of disease was within the first three weeks after the arrival of calves in the feedlots. Few other epidemiological descriptions (season, day of the week, geographical, age, sex, or breed) had been objectively described. The most common clinical and necropsy diagnoses were respiratory infections, often described as shipping fever. PMID- 17422727 TI - Isolation of Haemophilus agni from Six Alberta Ram Lambs with Septicemia. AB - Six ram lambs were submitted to a veterinary diagnostic laboratory for necropsy. Clinical signs included sudden illness or death with or without observed depression, reluctance to move, scours or fever. Gross findings and histopathology revealed evidence of bacterial septicemia. Haemophilus agni was isolated from brain, spleen, lung and lymph node. PMID- 17422728 TI - Fine needle aspiration biopsy of malignant tumors in dogs and cats: a report of 102 cases. AB - Fine needle aspiration biopsy is a fast and inexpensive technique well tolerated by animals. Its efficacy for the diagnosis of malignancy and determination of cellular origin was investigated for 97 malignant tumors and five benign lesions in 83 dogs and 19 cats. Histological examination of the lesions was performed in each case. Malignancy was detected by cytology in 69% of the malignant tumors. The cellular origin of the lesion was determined in 74% of the cases. Confirmation of malignancy and determination of cellular origin permitted an early diagnosis and prognosis. Since fine needle aspiration biopsy and cytology cannot definitely rule out malignancy, a surgical biopsy and histopathology should be done when malignancy is suspected clinically and not confirmed cytologically. Specimens of good quality, adequate collaboration between the cytologist and clinicians, and sufficient knowledge of pathology and cytology are basic requirements for obtaining good results with fine needle aspiration biopsy and cytology. PMID- 17422729 TI - Transmission and control of leptospiral infection in a beef cattle herd in southern alberta. AB - Leptospiral antibodies were detected in unvaccinated cattle on a 17 000 hectare ranch in the arid southeast region of Alberta. Antibody to serovar hardjo was present before the breeding season in 7% of 42 yearling bulls, 86% of 29 two year old bulls and 5% of 519 cows. Pomona antibody was confined to 3.7% of the cows. Bulls were treated once with dihydrostreptomycin, 25 mg/kg. Bulls and cows were vaccinated twice at a six week interval, with pomona-hardjo-gripptotyphosa bacterin before breeding and cows were revaccinated the next year. Leptospires were demonstrated in urine, kidney and spinal fluid of vaccinated and treated cattle. New infections occurred on range in vaccinates. Eighteen months after the last vaccination, hardjo and pomona antibody prevalences in cows were 3.6 and 3.2% respectively. A group of 250 seronegative cows on the same ranch were not vaccinated. They remained seronegative throughout the 2.5 years of the study. These cows, in contrast to infected groups, were excluded from pastures adjacent to perimeter herds and grazing leases, and they were bred by artificial inseminstion. Rotation through pastures in common with infected groups, and exposure to seropositive heat detector bulls, did not result in seroconversion in these cows. The study showed the potential of range bulls to amplify and transmit hardjo infection, limitations to the value of treatment and vaccination with available agents, and the potential of management practices to maintain an uninfected herd in close proximity to cattle carrying hardjo infection. PMID- 17422730 TI - Erythrocyte osmotic fragility testing and the prediction of canine malignant hyperthermia susceptibility. AB - A Doberman-German Shepherd cross-bred male dog, previously diagnosed as malignant hyperthermia susceptible, was mated to an unrelated nonsusceptible German Shepherd cross-bred female. The resultant litter was subjected to hematological, biochemical and erythrocyte osmotic fragility testing in an endeavor to predict the susceptibility of individuals to malignant hyperthermia. Laboratory evaluations were repeated at one year of age and the litter subjected to the halothane challenge test. No significant difference in erythrocyte osmotic fragility was found between malignant hyperthermia susceptible and nonsusceptible siblings at six weeks or at one year of age. Erythrocyte osmotic fragility, in both malignant hyperthermia susceptible and nonsusceptible animals, increased between six weeks and one year of age. Dantrolene sodium was an effective treatment for malignant hyperthermia in the dog when administered early in an episode and in adequate dosage. The initial sign of a malignant hyperthermia episode was a very rapid increase in end tidal partial pressure of carbon dioxide. This finding reinforces the value of capnographic monitoring in anesthesia. PMID- 17422731 TI - Ovariectomy, ovariohysterectomy and orchidectomy in rodents and rabbits. AB - This article describes the surgical procedures for the ovariectomy, ovariohysterectomy and orchidectomy in rodents and rabbits. The indications for each procedure are outlined and effective and safe anesthetics are described. Preoperative and postoperative care is also discussed. PMID- 17422733 TI - Nutrition. PMID- 17422732 TI - What is Veterinary Malpractice? PMID- 17422734 TI - Historical column: rinderpest and mainstream infectious disease concepts in the eighteenth century. PMID- 17422735 TI - The reindeer industry in Canada. PMID- 17422736 TI - Causal Inferences: An Epidemiologist's Viewpoint. PMID- 17422737 TI - Author's Reply. PMID- 17422738 TI - Cross-Canada disease report. PMID- 17422739 TI - Factor XI Deficiency in Canadian Holsteins. PMID- 17422740 TI - Detection of Actinobacillus pleuropneumoniae Infection in Pigs. AB - It is difficult to control the spread of porcine haemophilus pleuropneumonia caused by Actinobacillus pleuropneumoniae because there is no sensitive and specific way to accurately determine whether or not a pig herd is infected. This paper reports bacteriological and serological techniques used to detect A. pleuropneumoniae infection in pigs from a herd with endemic disease.The bacteria were isolated from the anterior nasal mucosa of grower pigs, but not from younger or older pigs. Bacteriological culture of several tissues from the respiratory tract showed that nine of ten young finishing pigs were infected, but culture of lung tissue from slaughtered hogs detected infection in only 39 of 288 (13.5%). Both cooler storage temperature and use of selective medium prolonged the time that lung tissue could be stored and the organism still recovered. An enzyme linked immunosorbent assay detected serotype-specific antibodies in serum of infected pigs. PMID- 17422741 TI - Mastitis Caused by Haemophilus somnus in a Dairy Cow. PMID- 17422742 TI - A reduction in litter size on an ontario Swine farm. AB - In a 200 sow herd, the litter size fell from an average of 10.5 pigs born alive per litter from January to June, to an average of 9.2 for the remainder of the year. Management changes during the first part of the year resulted in half the sows weaning litters at three weeks of age and half the sows weaning at four weeks of age instead of at six weeks as was previously done. The subsequent litter size was 9.3 pigs born alive per litter for three-week weaned sows compared to 10.0 for four-week weaning. The management of gilt breeding was also altered by the necessity to breed groups of 12 gilts in one-week periods of time and therefore a higher proportion of gilts may have been mated on their first estrus instead of their second or third estrus as had been the case. The difference in litter size of first parity sows between the first six months and the second six months was 1.1 pigs. Parvovirus infection may have been a factor contributing to the reduction in litter size amongst first parity sows; two groups of first parity sows experienced an increase in mummified piglets, a reduced far rowing rate, and smaller litter size. However, no attempt was made at diagnosing an infectious agent. Parity distribution remained relatively unchanged during the year and was not associated with the drop in litter size. PMID- 17422743 TI - A Survey of Eimeria spp. in Cattle in Central Alberta. PMID- 17422744 TI - Cryptosporidium antibodies in manitoba cattle: a pilot study using an indirect fluorescent antibody procedure. AB - An indirect fluorescent antibody test, using feces-derived oocysts as antigen, was used to detect antibodies to Cryptosporidium spp. in bovine sera in Manitoba. Antibodies were detected in 29 of 50 (58%) sera collected from animals of various ages on farms where calves had laboratory-confirmed cryptosporidiosis and in 76 of 186 (40.9%) sera collected at random from culled breeding stock. Serum antibody, presumably colostral in origin, did not appear to protect young calves from the infection. No geographic preference for the infection was demonstrated. PMID- 17422745 TI - Carbohydrates in cat diets: digestion and utilization. PMID- 17422746 TI - 3. Vital components of a computer system. PMID- 17422747 TI - Who publishes in the CVJ? PMID- 17422748 TI - How to Publish in the CVJ. PMID- 17422749 TI - Canine blastomycosis. PMID- 17422750 TI - Congenital skeletal deformity syndrome. PMID- 17422751 TI - To the CVMA. PMID- 17422752 TI - Why do research in zoological gardens? PMID- 17422753 TI - Distal peripheral polyneuropathy in a great dane. AB - A spayed female five year old Great Dane dog was diagnosed as having a chronic, progressive, symmetrical distal polyneuropathy and concurrent hypothyroidism. Axonal degeneration and segmental demyelination were evident in teased nerve fiber preparations. Clinical signs included hindlimb weakness and muscle atrophy of the head and distal limbs. Diagnosis was based on clinical, electrophysiological, and nerve and muscle biopsy findings. Thyroxine supplementation for one month was of no benefit. The etiology of the polyneuropathy was not established but several causes were considered. The extent of demyelination in our case was of greater magnitude than described in a previous report of a similar idiopathic distal symmetrical polyneuropathy in a Great Dane. PMID- 17422754 TI - Severity and nature of acidosis in diarrheic calves over and under one week of age. AB - A prospective study of the severity of dehydration and acidosis was carried out in 42 calves under 35 days of age presented for treatment of neonatal diarrhea. Clinically the mean level of dehydration was 8 to 10%. The plasma volume was 65% of that in the hydrated calf but the calves only gained 6.5% in weight during therapy.Calves under eight days of age often had a lactic acidosis. Blood pH was 7.118+/-0.026 (mean +/- 1 standard error), bicarbonate concentration 18.8+/-1.3 mmol/L, base deficit 11.4+/-1.7 mmol/L and lactate of 3.6+/- 0.06 mmol/L. Calves over eight days usually had a nonlactic acidosis. Blood pH was 7.042+/-0.021, bicarbonate 10.8+/-1.0 mmol/L, base deficit 19.5+/-1.2 mmol/L and lactate 1.2+/ 0.3 mmol/L. These values were all significantly different from those in younger calves.Over all calves there was a poor correlation between the severity of acidosis and dehydration(r=0.05). The severity of lactic acidosis was related to the severity of dehydration. Mean bicarbonate requirements to correct acidosis were calculated to be 200 mmol(17 g of sodium bicarbonate)and 450 mmol(37 g of sodium bicarbonate)in calves under and over eight days of age respectively. Both groups of calves required a mean volume of 4L of fluid to correct dehydration. PMID- 17422755 TI - An evaluation of nonsuppurative joint disease in slaughter pigs. AB - Fifty-two joints from pigs with nonsuppurative joint disease from a local abattoir were examined grossly, histologically, and microbiologically in order to establish macroscopic differences between degenerative arthropathy and arthritis due to an infectious organism. The joints were grouped grossly according to the type and severity of lesions of the synovial membrane and cartilage, and microscopically according to the severity of synovial membrane lesions. Osteochondrosis and Erysipelothrix rhusiopathiae were the most common causes of nonsuppurative joint disease in the joints examined. The major macroscopic differences between these two arthropathies were in the nature and severity of the synovial and cartilaginous lesions and involvement of the lymph node draining the diseased joint. Typically, in osteochondrosis, the changes are feathery hypertrophy of villi, focal full-thickness cartilage buckles, ulcers or flaps, and no change in the draining lymph node, whereas in Erysipelothrix- caused arthritis, the villous hypertrophy is severe and polypoid in nature, there is diffuse erosion of articular cartilage, and the draining lymph node is consistently hypertrophic and often cystic. PMID- 17422756 TI - Responses of pregnant ewes and young lambs to cold exposure. AB - The effects of cold stress were studied in pregnant ewes during the last three weeks of gestation and in their progeny during the first three days of life. In general, ewes were unaffected by treatment whereas changes were observed in the cold-stressed lambs. Cold-induced changes in lambs included physical weakness, depression, and poor nursing response. Serum concentrations of glucose and insulin were lowered whereas concentrations of blood urea nitrogen, alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, triglycerides, and cortisol tended to be higher in cold exposed lambs. The mortality rate was higher (40%) in cold-stressed lambs than in lambs kept at warmer temperatures (10%). At necropsy, cold-exposed lambs had reduced amounts of adipose tissue in perirenal areas, and extensive subcutaneous hemorrhages and edema in the distal portions of the thoracic and pelvic limbs. PMID- 17422757 TI - A case of marked and unresponsive obesity. PMID- 17422758 TI - Accidental sulfur poisoning in a group of holstein heifers. AB - Fourteen animals died or were euthanized after toxic levels of elemental sulfur were accidentally fed to a group of 120 Holstein heifers. Dehydration, rumen stasis, tachycardia, and diarrhea were seen along with metabolic acidosis, hypokalemia, and hypochloremia. The majority of deaths occurred from 3 to 10 days after the sulfur was fed to the heifers. Postmortem examination showed rumenitis, acute alveolitis, and renal tubular necrosis. The toxicity of ingested sulfur was attributed to the conversion of sulfur to hydrogen sulfide in the rumen. PMID- 17422759 TI - Adrenal degeneration associated with feather loss in a macaw. PMID- 17422760 TI - Acquired incarcerated inguinal hernia: a review of 13 horses. AB - The case records of 13 horses with acquired incarcerated inguinal hernia in January-August 1983, were reviewed. Nine cases were in stallions. The remaining four involved eventration 5-48 hours following castration. Ages ranged from 1-17 years. Horses showed a variable degree of colic. Bowel was felt to pass through the internal inguinal ring on rectal examination in most cases. The physical features of the scrotum varied considerably. Resection of ischemic jejunum and/or ileum was necessary in three horses. Two horses were euthanized at surgery (one with bilateral ischemic jejunum, one with bowel perforation), and a further horse on day 16 postsurgery following development of multiple adhesions. All stallions were castrated. Follow-up for 6-24 months (mean 12.7) disclosed that all ten discharged horses were alive and healthy (recovery rate 77%). PMID- 17422761 TI - Practitioner's Corner. PMID- 17422762 TI - Cross-Canada disease report. PMID- 17422763 TI - Telephone advice and the law. PMID- 17422764 TI - Pharmaceuticals and agriculture. PMID- 17422766 TI - Author's reply. PMID- 17422765 TI - Ectoparasites of cattle. PMID- 17422767 TI - Clinical diagnosis and surgical repair of a diaphragmatic hernia in a mare. PMID- 17422768 TI - Extension large colon resection in 12 horses. AB - Extensive resection (50-75%) of the large colon was performed in 12 horses. Indications for resection were: loss of viability due to large colon volvulus (seven), thromboembolic episode (three), impairment of flow of ingesta due to adhesions (one), or congenital abnormalities (one). The time required to correct the primary cause of abdominal pain and complete the resection ranged from 2.5 to 4.75 hours. Three horses had severe musculoskeletal problems postoperatively and were euthanized in the recovery stall. Four other horses were euthanized early in the postoperative period because of: further large colon infarction (two), ileus (one), or small intestinal problems (one). Five horses survived with no apparent nutritional or metabolic problems during two to three weeks of hospitalization. Clinical data were obtained from these horses from nine months to eighteen months postoperatively and revealed no clinical or clinicopathological abnormalities in four of them; the fifth horse exhibited diarrhea and weight loss four months postoperatively but responded to diet change. PMID- 17422769 TI - Congenital shoulder luxation in a fox terrier. PMID- 17422770 TI - Cystic nasal concha as a cause of unilateral nasal obstruction in a young bull. PMID- 17422771 TI - The Eyeworm, Thelazia skrjabini, in Cattle in Canada. PMID- 17422772 TI - Nematodirus battus in Canadian Sheep. PMID- 17422774 TI - Toxoplasmosis as a significant disease in man and animals with special reference to preventive measures by the farm community. PMID- 17422773 TI - Zoonoses on the family farm. PMID- 17422775 TI - Q Fever - human disease in ontario. PMID- 17422776 TI - Zoonoses involving family pets - bacterial and fungal pathogens. PMID- 17422777 TI - Chlamydiosis (psittacosis). PMID- 17422778 TI - Zoonoses in laboratory animals. PMID- 17422779 TI - Zoonoses in the slaughterhouse. PMID- 17422780 TI - Enteric pathogens and the food chain. PMID- 17422781 TI - Control of foodborne pathogens. PMID- 17422782 TI - Test your knowledge of zoonoses. PMID- 17422783 TI - Diary of a world war one veterinary officer. PMID- 17422784 TI - Practitioner's Corner. PMID- 17422785 TI - Cross-Canada disease report. PMID- 17422786 TI - 4. Computer hardware for a veterinary practice. PMID- 17422787 TI - Differing views on chuckwagon races. PMID- 17422788 TI - Animal abuse. PMID- 17422789 TI - Ivermectin toxicity in dogs and cats. PMID- 17422790 TI - Author's reply. PMID- 17422791 TI - Malignant hyperthermia in dogs. PMID- 17422792 TI - Author's reply. PMID- 17422793 TI - Author's reply. PMID- 17422794 TI - Clinical veterinary ethology in food animals. PMID- 17422795 TI - Commentary. PMID- 17422797 TI - Insurance. PMID- 17422796 TI - Veterinary contracts and the law. PMID- 17422798 TI - Cross-Canada disease report. PMID- 17422799 TI - Digestibility of pet foods. PMID- 17422800 TI - Dr. Tom Sanderson...living a rewarding experience. PMID- 17422801 TI - Clinical epidemiology. AB - Rational clinical practice requires deductive particularization of diagnostic findings, prognoses, and therapeutic responses from groups of animals (herds) to the individual animal (herd) under consideration This process utilizes concepts, skills, and methods of epidemiology, as they relate to the study of the distribution and determinants of health and disease in populations, and casts them in a clinical perspective.We briefly outline diagnostic strategies and introduce a measure of agreement, called kappa, between clinical diagnoses. This statistic is useful not only as a measure of diagnostic accuracy, but also as a means of quantifying and understanding disagreement between diagnosticians. It is disconcerting to many, clinicians included, that given a general deficit of data on sensitivity and specificity, the level of agreement between many clinical diagnoses is only moderate at best with kappa values of 0.3 to 0.6.Sensitivity, specificity, pretest odds, and posttest probability of disease are defined and related to the interpretation of clinical findings and ancillary diagnostic test results. An understanding of these features and how they relate to ruling-in or ruling-out a diagnosis, or minimizzing diagnostic errors will greatly enhance the diagnostic accuracy of the practitioner, and reduce the frequency of clinical disagreement. The approach of running multiple tests on every patient is not only wasteful and expensive, it is unlikely to improve the ability of the clinician to establish the correct diagnosis.We conclude with a discussion of how to decide on the best therapy, a discussion which centers on, and outlines the key features of, the well designed clinical trial. Like a diagnosis, the results from a clinical trial may not always be definitive, nonetheless it is the best available method of gleaning information about treatment efficacy. PMID- 17422802 TI - Interrelationships among Production and Clinical Disease in Dairy Cattle: A Review. AB - Epidemiological evidence is presented in order to answer two questions. The first question is: "Does high milk production put a cow at increased risk of disease?" The answer to this question seems to be "maybe" for milk fever, but "no" for most other common diseases (veterinary-assisted dystocia, retained placenta, metritis, cystic ovary, ketosis, left displaced abomasum, and mastitis). The second question is: "Is low milk production a consequence of disease?" For most diseases the answer is a cautious "yes". PMID- 17422804 TI - Le veterinaire et les grandes populations animales: l'exemple du porc. PMID- 17422803 TI - Physiopathologie et therapeutique de l'uterus en periode puerperale chez la vache laitiere: revue. AB - Accurate diagnosis and effective therapeutics of uterine pathologies in cattle represent some of the daily preoccupations for the veterinarians involved in herd health medicine. A rational approach requires a competence based on fundamental and clinical knowledge.The physiological aspects of the uterine involution in cattle are reviewed with emphasis about the in utero bacteriological dynamic and the endocrinological evolution during this period. The pathogenesis of uterine pathologies is discussed following new research results and new knowledge on the mechanisms of uterine defense. The value and limitations of the clinical diagnosis is briefly reviewed. The therapeutics of uterine pathologies is debated in relation with the pathophysiology of the uterine involution and with the rational of the traditional and new concepts of the treatment of the postpartum uterine pathologies. PMID- 17422805 TI - Approche clinique d'une porcherie: examen du batiment. AB - Environment is of primary importance to animal health and development. Ventilation is a problem in establishing an adequate environment, particularly in swine housing. In order for veterinarians to deal with this problem, a fundamental understanding of the building is necessary. The present article describes a clinical approach to ventilation diagnosis problems in the piggery. PMID- 17422806 TI - A new approach to managing health in Swine operations. AB - It is proposed that just as the stethoscope and thermometer are fundamental tools for individual medicine, production and health recording systems are fundamental tools for effective population medicine. Treatment and control of clinical diseases as the primary objective is no longer considered appropriate for livestock population. Disease in populations now describes a deviation between what is happening and what is expected to happen. This redefinition of disease implies that it is of multifactorial origin and thus a different problem solving approach must be implemented. Therefore, a swine enterprise must be considered as a system, a set of interdependent components continuously interacting to produce pork. As a system, it is characterized by certain properties: change, environment, counterintuitive behavior, drift to low performance, interdependency, and organization. A redefinition of diseases implies also that they are not only "treated" but managed. Management consists of planning, monitoring, evaluation, and analysis. For this process to be implemented successfully, a goal-directed recording system providing a farm-based infrastructure for problem solving is essential. Clinical problem solving (diagnosis) is thus based on epidemiological and demographic methods. PMID- 17422807 TI - A case study of respiratory disease in a veal calf operation. AB - An outbreak of respiratory disease occurred in a central Alberta veal operation, after production capacity had been increased fourfold. Mortality rate reached 24.6% despite agressive antibiotic therapy. A review of the records revealed a cyclical disease pattern in each room. Weekly cleaning of occupied calf rooms was correlated with the disease pattern. Aerosols generated by a high pressure sprayer appeared to trigger transmission of respiratory pathogens in malnourished neonatal calves.DISEASE OCCURRENCE DECREASED AND PROFITABILITY INCREASED SIXFOLD AFTER THE INTRODUCTION OF THE FOLLOWING MEASURES: 1) discontinuing the use of the high pressure washer in the occupied calf rooms, 2) feeding calves a better quality milk replacer with supplemental milk for the poorest calves, 3) sale-yard calf purchases were abandoned in favor of direct buying. PMID- 17422808 TI - The Management of Drug-induced Manipulation of the Estrous Cycle in Normal Cows and Heifers. AB - PART I PROSTAGLANDIN-INDUCED SYNCHRONIZATION OF ESTRUS IN BEEF CATTLE: Prostaglandin-induced regression of the mature cyclic corpus luteum in cows and heifers triggers a sequence of physiological events that results in a return to estrus in two to five days. There are several breeding management programs based on this premise. These programs range from attempts to synchronize estrus in entire herds with two injections of prostaglandin eleven days apart and breeding artificially, to simply shortening diestrus in responsive cattle with a single injection and bullbreeding. In this paper, several programs are discussed. No single program will be successful in all situations. Programs must be modified to fit each herd and its management. The factors that most commonly lead to program failure include inadequate nutrition, short postpartum interval, and mismanagement of heifers and first-calf heifers. PART II THE SYNCHRONIZATION OF ESTRUS IN EMBRYO TRANSFER RECIPIENTS USING VARIOUS SYNCHRONIZATION COMPOUNDS: Approximately 1800 recipient cows were synchronized for embryo transplants in three separate trials. The response rates, distribution of estrus, cull rates, and pregnancy rates of the synchronization products were compared. The pregnancy rates in prostaglandin-induced estrus in embryo transfer recipients were found to be no different from those in recipients that were used after natural noninduced estrus. The specific prostaglandin analog fenprostalene was tested for efficacy using various combinations of route of administration and antibiotic addition. There were no adverse reactions and neither the addition of oxytetracycline nor the route of administration had any effect on estrus rate, distribution, or pregnancy rates, which were not different from those achieved with the control prostaglandin analog cloprostenol. PART III TIMED BREEDING IN PROSTAGLANDIN SYNCHRONIZED DAIRY HEIFERS: Five groups of 20 to 40 Holstein heifers were treated with two doses of cloprostenol eleven days apart. The control groups were bred 12 to 16 hours after first seen in standing estrus. Treatment groups were bred at either 64 or 72 hours postinjection (with no detection) or at more than 24 hours postdetection of estrus. Optimum results were achieved when heifers were bred within 16 hours of first observed estrus or 72 hours after the second synchronizing injection with no detection of estrus. PMID- 17422810 TI - Public relations in veterinary medicine. PMID- 17422809 TI - Management of reproduction of sheep. PMID- 17422811 TI - Duncan McEachran in Retirement. PMID- 17422812 TI - Should veterinarians advertise? PMID- 17422813 TI - Lettre ouverte aux veterinaires du Quebec. PMID- 17422815 TI - Women veterinarians: pioneer and current. PMID- 17422814 TI - Ivermectin Dosage: a correction. PMID- 17422816 TI - Two sides of the coin: trend or tradition for advertising rules? PMID- 17422817 TI - The use of modern marketing strategies for the promotion of preventive medicine and herd health. PMID- 17422818 TI - Specifics of veterinary software. PMID- 17422819 TI - Cross-Canada disease report. PMID- 17422820 TI - The use of pharmaceutical products: an ongoing veterinary concern. PMID- 17422821 TI - Gastrointestinal upsets with high performance diets. PMID- 17422822 TI - Klebsiella oxytoca Septicemia due to Intravenous Administration of a Contaminated Amino-acid Solution to Three Horses. PMID- 17422823 TI - Serological Evidence of Coxiella burnetii Infection in Horses in Atlantic Canada. PMID- 17422824 TI - Canine sertoli cell tumor: a medical records search and literature review. AB - Seventeen cases of Sertoli cell tumor in dogs were found by searching medical records from 1971 to 1985. The average age of affected dogs was ten years, and most animals were of mixed breed. Most of the dogs were presented for reasons unrelated to the Sertoli cell tumor, however, most dogs had one or more clinical signs associated with Sertoli cell tumor. These signs included prostatic changes, alopecia and hyperpigmentation, bone marrow suppression, gynecomastia, and behavioral changes. The association of Sertoli cell tumor with cryptorchidism was obvious in these records, as ten of the dogs had one or both testicles undescended. Histopathological features of the tumors were variable and had no consistent correlation with metastatic potential. Some interesting comparisons can be made regarding these tumors in other domestic animals and in man. PMID- 17422825 TI - Potential inadequacy of the thyrotropin stimulation test in the diagnosis of canine hypothyroidism. PMID- 17422826 TI - Vaccination against coccidiosis in commercial roaster chickens. PMID- 17422827 TI - Isolation of two Mycoplasma species from the peritoneal cavity of a dog. PMID- 17422828 TI - Perceived Usefulness of the Collection of Subclinical and other Disease Entities Detected at Slaughter. AB - Based on a survey of 1820 Ontario pork producers and 16 veterinary members of the Ontario Swine Practitioners Association, condemnation/demerit data, while viewed as potentially valuable, are seldom, if ever, utilized in the provision of herd health programs. We found, however, that: 1) 89.4 percent (SD 1.04 percent) of producers and all veterinarians would use the disease information made available by a computerized system that would collect and report on subclinical and other disease entities detected during the slaughter of hogs; 2) Iesion diagnosis should be as detailed as possible and report on severity of disease; and 3) with the exception of those producers who ship larger numbers of hogs to market for which reports were wanted for each kill, monthly reports would be adequate.Written comments by producers expressed concerns about confidentiality of individual herd data, costs for the information/service, and veterinary expertise in the interpretation of findings. PMID- 17422829 TI - Five billion of us. PMID- 17422831 TI - Chuck wagon racing, greyhound racing. PMID- 17422830 TI - Veterinarians for amnesty. PMID- 17422832 TI - Canadian holstein cattle in "the warm heart of Africa". PMID- 17422833 TI - Meat and Right: Farming as if the Animal Mattered. PMID- 17422834 TI - Langford Laboratories - Biotechnology out of the Basement. PMID- 17422836 TI - The new college. PMID- 17422835 TI - Livestock Production and Veterinary Services in China - Keeping up with the Fourth Modernization. PMID- 17422837 TI - Partnerships and the law. PMID- 17422838 TI - Uterine rupture in a cat. PMID- 17422839 TI - Malignant catarrhal Fever in a five-month-old calf. PMID- 17422840 TI - Heartworm in dogs in Canada in 1986. AB - In late December 1986, 1224 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1986; 46% of them responded. Veterinarians reported that 150,989 dogs were blood-tested for microfilariae and 869 dogs were found with heartworm. Another 65 dogs were amicrofilaremic but diagnosed with heartworm disease and one was found with heartworm at necropsy to give the total number diagnosed in 1986 as 935 (0.62%).Heartworm was reported from Manitoba, New Brunswick, Ontario and Quebec, but most (810) of the cases were from Ontario. South-western Ontario continued to be the primary focus of the infection in Canada. There were 103 cases reported from Quebec, mostly from and around Montreal, and 21 cases from Manitoba, from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 33% of the cases were observed with clinical signs of heartworm disease and 81% had a history of not having left Canada. PMID- 17422841 TI - Equine viral encephalomyelitis in Canada: a review of known and potential causes. AB - Rabies, equine herpesvirus type I, and eastern and western encephalomyelitis viruses, known causes of equine neurological disease, are reviewed with emphasis on epidemiology, pathogenesis, clinical signs, and diagnosis.Several arboviruses known to be active in Canada and capable of producing neurological disease in humans (Powassan, St. Louis encephalitis, snowshoe hare, and Jamestown Canyon viruses) are discussed as potential causes of encephalomyelitis in horses. PMID- 17422842 TI - Maple syrup urine disease in five hereford calves in ontario. AB - Five newborn grade horned or polled Hereford calves in Ontario exhibited the clinical signs characteristic of hereditary neuraxial edema, namely, inability to rise, and hyperesthesia with extensor spasms upon tactile or auditory stimulation. Histologically, there was widespread spongy vacuolation in both white and gray matter in the brains of all five calves, especially in the midbrain, brain stem, and cerebellum. Amino acid analysis of formalin-fixed cerebral tissue of three calves demonstrated an amino acid composition consistent with a diagnosis of branched chain keto acid decarboxylase deficiency. This condition has been recently reported in Hereford calves in Australia; it is characterized by the delayed onset of neurological signs and severe lesions of status spongiosus, and has been considered analogous to maple syrup urine disease of children. PMID- 17422843 TI - A review of the pharmacology and clinical uses of ivermectin. AB - The avermectins were introduced in 1981 and constitute a potent new class of anthelmintic agents. They are naturally-derived products of microbial action displaying an exceptionally wide range of antiparasitic efficacy against internal and external parasites of domestic animals. This paper reviews their isolation and chemistry, mechanism of action, chemical efficacy and safety in cattle, sheep, swine, horses and dogs. PMID- 17422844 TI - Recovery of Mycoplasma spp. from the Reproductive Tract of the Mare during the Estrous Cycle. AB - The sites in the genital tract from which mycoplasmas could be recovered at various stages of the estrous cycle were studied in five Standardbred mares naturally infected with Mycoplasma. Mycoplasma equigenitalium and Mycoplasma subdolum were most frequently isolated from the clitoral fossa as compared to the vagina, cervix, and uterus. The lowest isolation prevalence was observed in the uterus. The recovery of Mycoplasma spp. from the clitoral fossa did not differ at any stage of the estrous cycle; however, recovery from the vagina, cervix, and uterus was variable during the cycle and more organisms were recovered on the day of ovulation than at any other time. From these results it was concluded that the clitoral fossa is the most likely "ecological niche" for Mycoplasma spp. in the mare. Ureaplasmas were not isolated. PMID- 17422845 TI - The Potential Economic Impact of an Outbreak of Foot-and-Mouth Disease in Canada. AB - The possibility of an outbreak of foot-and-mouth disease is of concern to Canada's livestock industry due to the resulting economic consequences. The primary economic impact of a foot-and-mouth disease outbreak would arise from the trade embargo placed on Canadian exports of animals and animal products to countries free of the disease. Agriculture Canada's Food and Agriculture Regional Model was used to estimate the economic impact of such a trade embargo. Two scenarios, a small and large outbreak, were simulated over a five year period (1986-90). The results indicate that even a small outbreak of foot-and-mouth disease would have serious economic consequences for the livestock sector with farm cash receipts declining by $2 billion. The largest impact would be on the pork sector followed by the beef sector. PMID- 17422846 TI - La mammillite herpetique bovine au Quebec. AB - BOVINE HERPETIC MAMMILLITIS IN QUEBEC: Bovine herpetic mammillitis is reported for the first time in Canada. It is a vesicular and ulcerative skin disease affecting the udder and teats of cows. It is caused by the bovine herpesvirus 2. The principal lesions consist of crusts that are found on the teats and may become complicated by secundary bacterial infection.Specimens collected from the lesions were used to differentiate the condition from pseudo-cowpox by serological tests, virus isolation and electron microscopy. Bovine herpetic mammillitis causes painful and therefore difficult milking which is followed by mastitis and an increased rate of culling. PMID- 17422847 TI - X chromosome mosaicism and infertility in a mare. PMID- 17422848 TI - Vegetarian dog foods. PMID- 17422850 TI - The CVMA International Development Committee. PMID- 17422849 TI - Canada's Experience with Foot-and-Mouth Disease. PMID- 17422851 TI - The OIE: a World Animal Health Organization. PMID- 17422853 TI - Certification of specialists. PMID- 17422852 TI - Guelph international development consultants. PMID- 17422854 TI - Ivermectin toxicity in small animals. PMID- 17422856 TI - Cold exposure of sheep. PMID- 17422855 TI - Author's Reply. PMID- 17422857 TI - Feline dilated cardiomyopathy. PMID- 17422858 TI - Methode simple pour la collecte des echantillons de sang chez les bovins a l'abattoir. PMID- 17422859 TI - The future of specialization: a report on the ontario veterinary association presidents' institute meeting. PMID- 17422860 TI - NEB Certification of Veterinary Specialists. PMID- 17422862 TI - Choosing a computer company. PMID- 17422861 TI - Veterinary Speciality Boards: Who Will be Guarding the Guards? PMID- 17422864 TI - Rex mears: commitment, integrity and humor. PMID- 17422863 TI - Cross-Canada disease report. PMID- 17422865 TI - The use of clenbuterol for the management of large animal dystocias: surgical corrections in the cow and ewe. AB - Clenbuterol (Ventipulmin(R) Solution) was included in the corrective management of dystocia cases requiring obstetrical surgery encountered in a Quebec rural practice. Data were collected on a total of 69 cases: 63 bovine and three ovine caesareans, and three bovine fetotomies. Ease of exteriorization of the gravid uterus and postoperative development of uterine adhesions were compared in two groups of bovine caesarean cases: a clenbuterol-treated group (Group A, n = 63) and a nontreated control group (Group B, n = 90). Clenbuterol produced a potent myometrial relaxant effect making its use beneficial in the correction of the bovine and ovine dystocias. It provided significantly improved uterine maneuverability (x(2) = 47.63, P < 0.001) and significantly reduced postoperative development of adhesions (x(2) = 14.05, P < 0.001) except in bovine caesarean cases with a high degree of myometrial distension. Neonatal and maternal health were not impaired and retention of fetal membranes was not increased. When used as directed, clenbuterol was a safe, reliable, and efficacious drug. PMID- 17422866 TI - Efficacy of Sulbactam, a B-lactamase Inhibitor, Combined with Ampicillin in the Therapy of Ampicillin-resistant Pneumonic Pasteurellosis in Feedlot Calves. AB - Two field efficacy studies, involving a total of 80 naturally infected feedlot calves, were conducted to compare the efficacy of sulbactam-ampicillin with that of penicillin-dihydrostreptomycin in the treatment of pneumonic pasteurellosis. Cultures from pretreatment nasal swabs were predominantly ampicillin/ penicillin resistant Pasteurella haemolytica. Clinical observations revealed that cattle treated with penicillin-dihydrostreptomycin responded poorly, whereas those treated with sulbactam-ampicillin responded promptly. Twenty-four hours after initiation of treatment, mean body temperatures of the calves in the sulbactam ampicillin groups had decreased by 2.1 degrees C, whereas, in the penicillin dihydrostreptomycin calves there was little change. The difference between the two treatments was statistically significant (P0.05). The rate of mortality did not differ significantly between the two groups. PMID- 17423141 TI - Colonic atresia in cattle: A prospective study of 43 cases. AB - This prospective study was initiated to document the success rate obtained in the treatment of colonic atresia in calves, identify factors that influence survival rate, and to report the histopathological appearance of the proximal blind end of the ascending colon. Forty-three calves with intestinal obstruction due to colonic atresia were admitted to the Ontario Veterinary College between September 1982 and May 1986. Parameters recorded prospectively in this study included age, breed, sex, history, vital signs, acid-base and electrolyte status, location of intestinal atresia, medical and surgical management, and outcome. The typical history and clinical signs included failure to pass meconium or feces, decreased appetite, and progressive depression and abdominal distension. The most common site of colonic atresia was the midportion of the spiral loop of the ascending colon (n = 25). Of the 43 calves, three (7%) were euthanized at surgery, 21 (49%) died in the hospital, and 19 (44%) survived and were discharged from the hospital. Four of the surviving calves died subsequent to discharge giving an overall long-term (mean 15.9 months) survival rate of 35%. No significant risk factors were identified, although experienced surgeons showed a trend towards increased survival rate. PMID- 17423142 TI - The Universal Transverse Mercator Code: A location code for disease reporting. AB - Since November 1987, all rabies specimen reports submitted by Agriculture Canada's District Veterinary Officers have required a new location code, the Universal Transverse Mercator Code (UTMC). In addition to the previously required entries for county, district, legal address and mailing addresses, the new code is set up for computer analysis and mapping. It is capable of pinpointing the origin of the specimen to within 100 meters anywhere in Canada that is covered by the National Topographic System 1:50,000 maps. Because of its 100 meter spatial resolution, the code is of great interest to those studying the occurrence and spread of rabies. The code will also be important in the detailed planning and evaluation of the Ontario rabies control scheme, scheduled for 1988. Agriculture Canada anticipates that the UTMC will also be used for reporting other animal diseases as well as for emergency disease reporting. PMID- 17423143 TI - A retrospective study of dystocia-related vertebral fractures in neonatal calves. AB - Forty-seven cases of neonatal vertebral fractures/luxations occurred in a 21 year period (1967-1987). All of the fractures were located between the 11th thoracic vertebra and the fourth lumbar vertebra; 77% occurred at the thoracolumbar junction. All but one case was associated with a forced extraction, either unspecified (53%), mechanical (28%), or manual (17%).A weak calf or continuous recumbency since birth was the major clinical sign. Hemorrhage around the kidneys, adrenal glands, and in perivertebral muscles was a consistent necropsy finding and a useful indicator that a thoracolumbar fracture was present. In addition to the vertebral fracture, the prominent necropsy findings were subdural and epidural hemorrhage, myelomalacia, spinal cord compression or severed spinal cord, and fractured ribs. All of the calves died or were euthanized without regaining locomotory function. PMID- 17423144 TI - Nasal amyloidosis in two Quarter Horses. PMID- 17423145 TI - Generalized subcutaneous emphysema in a dog. PMID- 17423146 TI - Efficacy of a pasteurella haemolytica vaccine/bacterial extract in the prevention of bovine respiratory disease in recently shipped feedlot calves. PMID- 17423147 TI - Epizootiology and pathology of avian tuberculosis in chickens in Saskatchewan. PMID- 17423148 TI - Cryptosporidium infection associated with proliferative enteritis (wet tail) in Syrian hamsters. PMID- 17423149 TI - Alberta. Mastitis associated with suspected Nocardia sp. PMID- 17423150 TI - Ontario. Recent gentamicin-resistant turkey Salmonella isolates. PMID- 17423151 TI - Saskatchewan. Ammoniated forage poisoning of cattle (;bovine bonkers'). PMID- 17423152 TI - British Columbia. Dinoseb toxicity in dairy heifers. PMID- 17423153 TI - Syncope in a retriever. PMID- 17423154 TI - Coprophagia: Food for thought. PMID- 17423155 TI - Veterinary locum tenens and the law. PMID- 17423156 TI - Do high-fat dog foods predispose dogs to pancreatitis? PMID- 17423158 TI - Are readers satisfied with the CVJ? PMID- 17423157 TI - The discovery of Chiron's cave, a prehistoric school of medicine for animals and humans. AB - Chiron the centaur, mythical inventor of medicine for animals and humans, and emblem of numerous veterinary associations worldwide, is possibly an historic personality of the prehomeric period (ca 1300 B.C.) Only 600 to 700 years later did the legend form that centaurs (in antique Greek: "hunters on horseback of the wild bull") were wild creatures, half horse and half man, and associated with uncontrolled libido and its socially destructive forces. According to tradition, literary as well as folkloric, Chiron taught his medicine to the ruling class of his time in a specially prepared cave in the Pelion Mountains (=the healing mountains) of Eastern Thessalia. Following leads from ancient authors and local traditions, a cave fitting the location and description was discovered in 1981 by the senior author. Details about its location, and observations about its surroundings, including ancient and modern place names indicate once more that Chiron, the centaur, stands for an historic personality. PMID- 17423159 TI - Veterinary ophthalmology in Canada. PMID- 17423160 TI - Servet: A personal contribution by all veterinarians to the hungry world. PMID- 17423162 TI - Veterinary consultation in Nicaragua. PMID- 17423161 TI - A survey of the readership of CVJ. AB - The results of a mail survey of a stratified random sample of Canadian Veterinary Journal (CVJ) subscribers are presented. The survey provided data about the level of satisfaction and amount of interest in journal articles, and associations between subscribers' willingness to purchase a subscription and area of professional activity, as well as with a number of readership variables.The readership indicated a general satisfaction with the CVJ. Most articles and subject categories were widely read, and satisfied the majority of readers. Subscribers indicated that the CVJ should assume a larger role in continuing education, and the willingness to purchase a subscription was associated with the subscriber's need for continuing education having been met.Unwillingness to purchase a subscription was associated with small animal professional activity and the opinion that too little emphasis in the journal was placed on small animal related articles and material useful in private practice.Recommendations were to increase the content of small animal subject material and to include more material of a practical nature in all subject categories. PMID- 17423163 TI - The decline of large animal practice? PMID- 17423164 TI - Retained fetal membranes in the mare: A retrospective study. AB - A retrospective study of 3456 deliveries was conducted from the records of four Standardbred broodmare farms where mares were bred by artificial insemination and maintained under close veterinary supervision. Retained fetal membranes (RFM) were observed in 10.6% of the deliveries. Retained fetal membranes occurred more frequently (p < 0.05) after dystocia and in mares which had RFM the previous year. Retained fetal membranes after normal foaling had no significant effect on the reproductive performance (pregnancy rate, pregnancy loss rate, or foaling rate), nor on the general health of the mares, regardless of the duration of RFM (3 to 144 hours). Postfoaling laminitis was not observed. Oxytocin therapy of mares with RFM starting at two hours postpartum significantly reduced the incidence of RFM >/= 8 hours. Mares with RFM which had received intrauterine antimicrobials between foaling and first breeding had a foaling rate similar to mares with RFM which had not received intrauterine therapy. PMID- 17423165 TI - A descriptive study of mortality at the Kortright Waterfowl Park: 1982-1986. AB - A five year survey of mortality in an Ontario waterfowl park was conducted. The highest mortality rate was in birds under seven weeks of age, with infectious disease most frequently diagnosed. In older birds, amyloidosis and gout were most common. Associations were found between the presence of amyloid and the diagnoses of gout, enteritis, and arthritis. Specific data collection requirements for studies of this type, and the difficulties involved, are outlined. Recommendations for increasing productivity, including the implementation of sanitation programs, and improvements to the record-keeping system, are made. PMID- 17423166 TI - Facteurs de risque nutritionnels de la pathologie hepatique dans les troupeaux bovins laitiers en France. AB - Nutritional risk factors for liver damage were surveyed in Black-Pied French dairy herds using serum glutamic dehydrogenase and gamma glutamyl transferase levels to monitor liver dysfunction.The 34 herds chosen were free of parasitic liver injuries. Feeding systems, nutritional blood parameters and production characteristics are analyzed. Two risk factors appeared to be involved in hepatic injury: feeding of fresh rape (Brassica napus) and high levels of serum urea (>/=6.64 mmol/L) associated with grass pasture or dietary nonprotein nitrogen with corn silage. PMID- 17423167 TI - Pneumonia associated with eugonic fermenter-4 bacteria in two Chinese leopard cats. PMID- 17423168 TI - Birth of a calf from bovine oocytes matured and fertilized in vitro. PMID- 17423169 TI - Blindness caused by hypovitaminosis A in feedlot cattle. PMID- 17423170 TI - Hypocalcemic seizures in an African grey parrot. PMID- 17423171 TI - Unilateral nephrectomy for treatment of chronic hematuria due to nephrolithiasis in a colt. PMID- 17423172 TI - Insulin treatment and development of anti-insulin antibodies in a horse with diabetes mellitus associated with a functional pituitary adenoma. PMID- 17423173 TI - Alberta.Trypanosoma theileri in cattle of central Alberta. PMID- 17423174 TI - Ontario. Equine arteritis virus isolated from a Standardbred foal with pneumonia. PMID- 17423176 TI - Alberta. Fatal dietary imbalance in horses. PMID- 17423175 TI - Saskatchewan. Selenium toxicity in suckling pigs. PMID- 17423177 TI - Pleural effusion in a dog. PMID- 17423179 TI - Multiple mucocutaneous lymphosarcoma in a dog. PMID- 17423180 TI - Giant cell hepatitis in four aborted foals: A possible leptospiral infection. PMID- 17423181 TI - Oxalate nephropathy associated with seizures in mink. PMID- 17423182 TI - Canine mandibular fracture repair using a technovit transfixation splint. PMID- 17423183 TI - Concurrent ileocecal and cecocolic intussusception in a horse. PMID- 17423184 TI - Haemophilus somnus myocarditis in a feedlot steer. PMID- 17423185 TI - Saskatchewan. Avian cholera epizootic in wild ducks. PMID- 17423187 TI - Western Canada. Rabies diagnoses, january 1 - june 30, 1988. PMID- 17423186 TI - Ontario. Rabies in a squirrel. PMID- 17423188 TI - Alberta. Levamisole toxicity in lambs. PMID- 17423190 TI - Lite diets. PMID- 17423189 TI - The problem of destructive scratching by cats. PMID- 17423192 TI - The changing role of the government vet. PMID- 17423193 TI - Editor's reply. PMID- 17423194 TI - More on the"information explosion". PMID- 17423195 TI - International development: A new beginning. PMID- 17423197 TI - Casting votes for Animal Health Week. PMID- 17423196 TI - Skin biopsy: Revelations and limitations. PMID- 17423198 TI - Effectiveness of an ivermectin liquid formulation given by nasogastric tube against strongyles in horses. AB - Twenty horses were treated with ivermectin either by nasogastric tube with a liquid formulation for sheep or per os with a paste formulation for horses at a dosage of 200 mug/kg of body weight. Fecal samples were collected from these horses and from ten untreated horses at the time of treatment and every 2 wk thereafter for up to 10 wk. The samples were examined for nematode eggs using the Cornell-McMaster dilution and the Cornell-Wisconsin Double Centrifugation procedures.There were no signs of toxicosis in horses treated with ivermectin. Strongyle eggs were found in the feces of all horses before treatment. Subsequently, they were found in untreated horses, but not in treated horses at 2 wk nor in most of them for up to 8 wk after treatment. At 10 wk most of these horses had strongyle eggs in their feces, but in general fewer than at pretreatment. PMID- 17423199 TI - Immune response to an Actinobacillus pleuropneumoniae vaccine in swine. AB - Piglets vaccinated with an inactivated tetravalent vaccine (Pleurovac 4) against Actinobacillus pleuropneumoniae serotypes 1, 2, 5 and 7, produced circulating antibodies after a first intramuscular injection and showed an anamnestic reaction after a second. The rise in antibody levels in vaccinated piglets was statistically significant when compared with those of the control group. The administration of 1 or 2 mL of vaccine did not lead to significantly different antibody levels. The specificity of the immune response is demonstrated by an increase in titer to all four serotypes in pigs given the tetravalent vaccine, but an increase in titer to only two serotypes in pigs given a bivalent vaccine (Pleurovac). PMID- 17423200 TI - An outbreak of cysticercosis in feedlot cattle. AB - An outbreak of cysticercosis (infestation with the larvae of Taenia saginata) occurred in feedlot cattle in Ontario in 1986. Two hundred and thirty-three of 271 steers were confirmed histologically to be positive for cysticerci. Nineteen (8.2%) animals had viable cysticerci, 87 (37.3%) had degenerated cysticerci, 77 (33.0%) had mineralized cysticerci, and 50 (21.5%) steers had lymphoid granulomas consistent with cysticercosis. Three viable cysticerci were partly evaginated and one degenerate cysticercus was fully evaginated. PMID- 17423201 TI - Acquired hydrocephalus and hydromyelia in a cat with feline infectious peritonitis: A case report and brief review. AB - A one-year-old domestic long-haired cat was referred to the New York State College of Veterinary Medicine because of acute onset of paraparesis and hyperesthesia associated with trauma. Myelography and cerebrospinal fluid analysis revealed severe hydromyelia and myelitis, respectively. The definitive diagnosis of feline infectious peritonitis was made by histological examination at necropsy. Lesions were confined exclusively to the brain and spinal cord. Partial occlusion of the third and fourth ventricles with pyogranulomatous debris caused hydrocephalus and subsequent hydromyelia. The hydromyelia may have been the primary means of compensation for the hydrocephalus, thus masking subclinical disease. PMID- 17423203 TI - Euthanasia with nitrogen. PMID- 17423202 TI - Authors' reply. PMID- 17423204 TI - One animal among many: Veterinarians in the global community. AB - Evidence from chemistry, physics, the biological sciences and the earth sciences is now accumulating that the earth functions as a living organism, with all the implications of circulatory and homeostatic systems which that implies. As veterinarians, we are more than simply handmaidens to agribusiness, the academic, pet and food industries, or government regulatory agencies. While our jobs may be specific, dependent, and circumscribed, our vocation can be nothing less than the health-care of all animal life - and hence by direct implication, the care of the earth, as a living organism, itself. PMID- 17423205 TI - Alleviation of postanesthetic hypoxemia in the horse. AB - This study was designed to investigate the effect of the nasotracheal insufflation of oxygen at a flow rate of 15 L/min on the arterial partial pressure of oxygen during the recovery period following inhalation anesthesia in the horse. It has been stated that this is a suitable flow rate to prevent postoperative hypoxemia but without any experimental evidence to support those statements. Horses being used for the study of healing of cartilage were anesthetized on two separate occasions. Following one period of anesthesia they were allowed to recover breathing room air, and following the other period of anesthesia oxygen was insufflated into the trachea at 15 L/min throughout the recovery period. This permitted each horse to act as its own control and allowed statistical analysis using Student's t-test for paired samples.The insufflated horses had a higher arterial partial pressure of oxygen during the recovery period than did the noninsufflated horses (p < 0.05). PMID- 17423206 TI - Bovine lead poisoning in Alberta: A management disease. AB - Lead poisoning was the most common toxicosis diagnosed in cattle by Alberta Animal Health Laboratories between 1964 and 1985 (n = 738 cases, x = 33.5 cases per year). Seasonal variation in incidence was evident, and occurrence was frequently associated with change in housing or pasture. Discarded batteries or used crankcase oil were implicated in more than 80% of cases for which the source of lead was determined.Pulmonary congestion, marked congestion and hemorrhage of thymus and heart, and presence of oil or lead particles in the ingesta were the most common postmortem findings. Eighty-six percent of cases were confirmed by elevated lead levels in tissues.Lead poisoning represents a significant, unnecessary loss to producers. Increased producer awareness and improved waste management on farms could significantly reduce the incidence of lead poisoning in cattle. PMID- 17423208 TI - Where are the authors hiding? PMID- 17423207 TI - Open wound management for treatment of postoperative infections in eight dogs. PMID- 17423210 TI - Rapid determination of equine hemoglobin concentration with the Spencer hemoglobinometer. PMID- 17423209 TI - Sensibilite a l'halothane dans quelques elevages de porcs Landrace au Quebec. PMID- 17423211 TI - The anthelmintic effectiveness of oxfendazole when administered by intraruminal injection to beef cattle. PMID- 17423212 TI - Calcinosis cutis secondary to percutaneous penetration of calcium carbonate in a Dalmatian. PMID- 17423213 TI - Hyperadrenocorticism in association with an adrenocortical adenoma in a pet ferret. PMID- 17423214 TI - Orthodontic correction of deviated canine teeth in the dog. PMID- 17423215 TI - Failure of a seropositive bull to transmit Leptospira interrogans serovar hardjo infection to heifers. PMID- 17423216 TI - Quebec. Colonic spirochetosis in piglets. PMID- 17423218 TI - Saskatchewan. Chronic pleuritis in a horse. PMID- 17423217 TI - Ontario. Diseases of imported marine tropical fish. PMID- 17423219 TI - Alberta. Inherited renal disease in lambs. PMID- 17423221 TI - Exercise in electrocardiography. PMID- 17423220 TI - AIDS and the veterinary profession. PMID- 17423222 TI - The case of the dumb gilts. PMID- 17423223 TI - Low ash cat foods: The role of magnesium in feline nutrition. PMID- 17423224 TI - Authors' reply. PMID- 17423225 TI - Bad press for bats. PMID- 17423226 TI - Important to have a Retirement Savings Program? PMID- 17423228 TI - A rapid stick test for the diagnosis of bovine leukemia virus infection from serum or milk. PMID- 17423229 TI - The future of food animal practice. PMID- 17423230 TI - A call for changes in veterinary education. PMID- 17423232 TI - Future of food animal practice. PMID- 17423231 TI - The proposed new cruelty to animals law - Implications for veterinarians. PMID- 17423233 TI - High sulfur related thiamine deficiency in cattle: A field study. AB - Following development of polioencephalomalacia in one of 105 cattle in a farm in southeastern Saskatchewan, a study was initiated to monitor thiamine (B(1)) and copper (Cu) status, and to evaluate interactive nutritional factors which may have been responsible for this occurrence. It was evident that a combination of high sulfur (S) and low Cu intake was responsible for the depletion of blood B(1) and plasma Cu. Supplementation with trace minerals alone resulted in a significant (p<0.05) improvement in both B(1) and Cu status of the herd. We recommend that herds exposed to high intakes of S be supplemented with Cu up to 50 mg/kg feed dry matter to alleviate potential deficiencies of B(1) and Cu. PMID- 17423234 TI - Optimal time postbreeding for pregnancy examination in dairy cattle. AB - The objective of this study was to determine the relationship between calving interval and the interval from artificial insemination to pregnancy examination. Cows examined for pregnancy at 30 or more days postbreeding were followed to determine the calving interval for cows examined at sequential intervals postbreeding (e.g. 42-44 days, 45-47 days postbreeding). Cows included in the study were from five large dairies in California, were presented for pregnancy examination from December of 1982 to April of 1983, and had a calving interval of from 290 to 530 days. All palpations were done by one of two experienced veterinarians. Calving intervals were significantly lower for cows pregnancy examined from 51-56 days postbreeding than for cows examined earlier or later. PMID- 17423235 TI - Heartbase tumor and pericardial effusion in a dog. AB - Pericardial effusion and congestive heart failure occurred in a ten-year-old male Bouvier des Flandres dog. Signs included weakness, exercise intolerance and ascites. Echocardiography identified a heart base mass as a possible cause of the pericardial effusion. A large quantity (650 mL) of bloody fluid was removed by pericardiocentesis and was characterized at cytology as a nonseptic sanguineous exudate. There was no cytological evidence of neoplasia in the effusate. Pericardiocentesis caused dramatic clinical improvement and resolution of the signs of congestive heart failure. Surgical excision was attempted but the mass could not be resected since it invaded the entire dorsal wall of the right atrium as well as part of the aortic root. The cellular origin of the heartbase tumor could not be determined. PMID- 17423236 TI - Morphological study of bacterial pneumonia of feedlot cattle: Determination of age of lesions. AB - Lungs from 48 feedlot cattle that had died from bacterial pneumonia were examined grossly and microscopically. Criteria based on microscopic lesions were adopted to age these pneumonias. In 38 cases, pneumonic lesions were of relatively uniform age throughout the affected tissue. In eight other cases, the presence of older lesions confined to one or two lobes suggested a previous episode of pneumonia. The aging criteria adopted were in agreement with the duration of the observed clinical signs in 26 cases. In 13 other cases, the pneumonia was estimated to be of longer duration than suggested by the history, whereas in the remaining nine cases, it was estimated to be more recent. Areas of tan discoloration of the parenchyma surrounded by white or yellow borders were considered the best areas to examine microscopically since they offered the best chances of revealing necrosis and fibrosis, the main lesions used to age the pneumonia. PMID- 17423237 TI - Comparative cost-effectiveness of ivermectin versus topical organophosphate in feedlot calves. AB - A study was conducted in a commercial feedlot in western Canada to evaluate the impact of treatment with ivermectin versus a topical organophosphate on growth rate and feed efficiency in calves entering a feedlot at an average 275 kg liveweight.A total of 9527 head of cattle was used. Variables measured included average daily gain, average days occupation, and feed conversion.Ivermectin treated animals gained an average 0.08 kg per day more than those treated with topical organophosphate. In addition, they required an average 0.23 kg less feed/kg gain. Based on an average 227 kg of gain, this would result in 11 fewer days occupation and 52.3 kg less feed for ivermectin treated animals. This equaled a net benefit of $7.04 per head over treatment costs for ivermectin treatment versus topical organophosphate. PMID- 17423238 TI - Monensin poisoning in horses - an international incident. AB - Several hundred Michigan horses were accidentally exposed to varying levels of monensin. Severity of effects was proportional to the level of feed contamination; sudden death resulted on at least two premises. Acute signs of cardiovascular impairment occurred on one premises having received feed containing over 200 grams of monensin per tonne. Gross and histological postmortem lesions consisted of acute myocardial necrosis. Although only circumstantially confirmed, investigations led to the suspicion that the source of poisoning was a ration formulation error in a feedmill in southwestern Ontario. Concern over possible undetected heart damage in exposed horses led to clinical monitoring on one farm over a period of several months. Electrocardiographic and serum enzyme monitoring were used soon after the incident to implicate exposure in some horses; they were poor prognostic indicators. Applicable legislation, the cooperative role of government departments, and legal implications relative to potential prosecution and lawsuits arising from sale of contaminated feed between Canada and the USA are summarized. PMID- 17423239 TI - Morbidity and mortality rates - handle with care. PMID- 17423240 TI - Benzimidazole resistance in horses in western Canada. PMID- 17423241 TI - Surgical treatment of an esophageal laceration in a calf. PMID- 17423243 TI - Alberta. Avian salmonellosis. PMID- 17423242 TI - Ontario. Encephalomyocarditis virus outbreak among suckling pigs. PMID- 17423244 TI - Alberta. Organophosphate toxicity in dairy cattle. PMID- 17423245 TI - An outbreak of keratoconjunctivitis (pinkeye) in sheep. PMID- 17423246 TI - Otobius megnini (spinous ear tick) in a dog. PMID- 17423247 TI - Exercise in electrocardiography. PMID- 17423248 TI - What is your Diagnosis? PMID- 17423249 TI - Feed energy (caloric density): Determination and significance in pet foods. PMID- 17423250 TI - The prospect before us. PMID- 17423251 TI - Monaco, november 1989. PMID- 17423253 TI - Do we need training in management? PMID- 17423254 TI - The situation in Nicaragua. PMID- 17423255 TI - Canine hyperthermia in December. PMID- 17423256 TI - Flea products and environmental protection. PMID- 17423257 TI - Authors' reply. PMID- 17423258 TI - The history of veterinary medicine. PMID- 17423259 TI - Effect of large palpable ovarian follicles on response to prostaglandin administration in dairy cows with corpora lutea. AB - We examined the response to exogenous prostaglandin F2alpha in cattle with or without palpable structures believed to be ovarian follicles. All animals had ovarian structures diagnosed by palpation as corpora lutea. The cows were placed into two groups: those with follicles which were estimated by the palpators to be 60 cows) and herds with a history of an outbreak prior to 1987 had increased chances of an outbreak occurring in 1987-1988.These results are compatible with an infectious cause for winter dysentery. PMID- 17423475 TI - Aspects of the epidemiology of anthrax in Wood Buffalo National Park and environs. PMID- 17423476 TI - Association of weak estrual signs and irregular estrous cycles with hypothyroidism in a bitch. PMID- 17423477 TI - The influence of a premating injection of gonadotrophin-releasing hormone on sow and gilt fertility. PMID- 17423478 TI - Postcastration eventration in eight horses. PMID- 17423480 TI - Canada. Nocardia spp. mastitis in Canada. PMID- 17423479 TI - Nasal papillary adenoma in a goat. PMID- 17423481 TI - Alberta. Spinal poliomalacia in weaned beef calves. PMID- 17423482 TI - Western Canada. Paramyxovirus infection in pigeons. PMID- 17423483 TI - Saskatchewan. Pelodera dermatitis in a dog. PMID- 17423484 TI - Equine embryo transfer. PMID- 17423485 TI - Eosinophilic enteritis in an Irish Setter dog. PMID- 17423486 TI - Dealing with canine obesity. PMID- 17423488 TI - Feedtrough dirt as a source of Clostridium botulinum type C intoxication in a group of farm horses. AB - Four horses from the same farm developed clinical signs of botulism during the winter months; three of these horses died. One horse survived an initial attack and recovered over a three-week period, but died during a second attack. The horse that survived took six weeks to recover. Clinical and postmortem examination ruled out other causes of disease. Confirmation of the diagnosis was made by isolation of Clostridium botulinum type C toxin from the dirt in the bottom of an oak feedtrough used by all horses, and from the colonic contents of one of the horses that died. To our knowledge, this is the second case of C. botulinum type C intoxication reported in horses in North America. In both cases, soil and sand near aquatic environments were identified as the source of toxin. PMID- 17423489 TI - A study of hepatic lesions in broiler chickens at processing plants in Saskatchewan. AB - A detailed descriptive study was done on broiler chickens with abnormal livers found at processing. Two syndromes were evident: those birds with enlarged, pale, firm livers, designated hepatosis, with Clostridium perfringens type A often isolated; and ascitic birds with cobblestone-appearing livers. Livers with hepatosis had marked proliferation of bile ducts; ascitic livers had normal architecture. Hearts from birds with ascites had elevated right ventricular/total ventricular weight ratios, whereas hearts from normal birds and birds with hepatosis did not. Two isolates of C. perfringens produced necrotic enteritis in experimental birds; an attempt to reproduce hepatosis with these isolates was unsuccessful. PMID- 17423490 TI - Ectopic ureter in the horse: Three cases and a review of the literature. AB - Ureterovesicular anastomosis resulted in resolution of the clinical signs of urinary incontinence in three horses with unilateral ectopic ureter. Follow-up of two of the horses ten months and three years later indicated no further urinary tract problems; the third horse died four days after surgery from intestinal infarction.Diagnosis can be readily confirmed by antegrade or retrograde ureterography, or endoscopic visualization of the ectopic ureteral openings. Nephrectomy appears indicated in cases of unilateral ectopic ureter with associated ipsilateral urinary tract infection or hydronephrosis, providing the contralateral kidney has normal function. Ureterovesicular anastomosis appears to be indicated in the management of unilateral ectopic ureters in the absence of ascending urinary tract infection or hydronephrosis, and in cases of bilateral ectopic ureters. PMID- 17423491 TI - A congenital defect resembling Hereford syndrome I in Alberta. PMID- 17423492 TI - Hypocalcemia associated with muscular weakness and recumbency in beef cows in western Saskatchewan. PMID- 17423493 TI - Immunoglobulin concentrations in commercially available colostrum supplements for calves. PMID- 17423495 TI - Ontario. Bulk tank survey of dairy farms in Ontario. PMID- 17423494 TI - Concurrent lymphoma and salmonellosis in a cat. PMID- 17423496 TI - Saskatchewan. Association of Actinobacillus ureae (Pasteurella ureae) with porcine abortion. PMID- 17423497 TI - Alberta. Salmonella arizonae in sheep. PMID- 17423498 TI - Exercise in electrocardiography. PMID- 17423499 TI - Diagnosing dominance aggression. PMID- 17423500 TI - A propos de l'obesite chez le chien. PMID- 17423501 TI - Volunteerism and the delivery of CVMA program. PMID- 17423502 TI - CVMA insurance program. PMID- 17423504 TI - Lest we forget! PMID- 17423505 TI - Animal rights activists. PMID- 17423506 TI - Author's reply. PMID- 17423507 TI - Clinical and laboratory findings of a bleeding disorder in Simmental cattle. AB - Eight Simmental cattle were presented to the Western College of Veterinary Medicine over the past decade with historic or current prolonged episodes of bleeding. Spontaneous epistaxis, superficial hematomas, and prolonged bleeding following injury or minor surgery (such as tattooing) were frequent observations. Animals presented because of prolonged hemorrhage were anemic and debilitated. Compensatory erythropoiesis was compromised in these animals by depletion of iron stores due to chronic blood loss. Screening tests to assess coagulation were normal as were numbers and morphology of platelets. In vitro platelet aggregation was abnormal in all affected cattle. Three animals were euthanized due to uncontrollable hemorrhage but the precise source of hemorrhage was undetectable. These findings are consistent with bleeding due to a platelet function disorder which has been described previously in Simmental cattle. PMID- 17423508 TI - Serum progesterone determination as an aid for pregnancy diagnosis in goats bred out of season. AB - Nubian does (n = 12) were bred by artificial insemination after induction of estrus with medroxyprogesterone acetate impregnated vaginal sponges and pregnant mare serum gonadotrophin injections during the anestrous season. Pregnancy status was predicted from serum samples collected 21 days following the last breeding and analyzed using 1) a commercial bovine milk progesterone enzyme immunoassay test (EIA), and 2) a radioimmunoassay progesterone (RIA) test. Both tests detected nonpregnancy (EIA 100%, RIA 80%) more accurately than pregnancy (EIA 66%, RIA 75%). Commercial bovine progesterone EIA kits have potential as rapid, inexpensive screening tests for nonpregnant does bred out of season. PMID- 17423509 TI - Resolution of pyothorax in a deer by a thoracotomy, drainage and lavage via tube thoracostomy. PMID- 17423510 TI - Multiple congenital skeletal deformities in an Arabian foal. PMID- 17423511 TI - Ammonium urate urolith resulting in hydronephrosis and hydroureter in a dog with a congenital portosystemic shunt. PMID- 17423512 TI - Ontario. Mortality caused by improper vaccination techniques in chickens. PMID- 17423514 TI - Alberta. Leech infestations in trout. PMID- 17423513 TI - Saskatchewan. An unusual cause of nitrate poisoning in cattle. PMID- 17423515 TI - Update on butorphanol tartrate: use in small animals. PMID- 17423516 TI - A veterinarian's guide to the correction of dominance aggression. PMID- 17423517 TI - Carbohydrates in the nutrition of the dog. PMID- 17423520 TI - Animal rights. PMID- 17423519 TI - Dealing with substance dependence. PMID- 17423521 TI - Quality assurance and continuing competence. PMID- 17423522 TI - Electronic communication with veterinarians: An update on VetNet and Exten. PMID- 17423524 TI - Pleura, pleurae, plural - In praise of exactitude. PMID- 17423523 TI - Five field trials on the efficacy of a bovine respiratory syncytial virus vaccine. AB - Five field trials evaluated whether immunization of beef cattle prior to weaning, at weaning, or immediately upon arrival at the feedlot with a commercial bovine respiratory syncytial virus (BRSV) vaccine would reduce subsequent treatment for respiratory disease.Bovine respiratory syncytial virus vaccination was associated with a significant (p<0.05) reduction in treatment rate in one of three groups of calves immunized prior to weaning (-12%) and in calves immunized upon arrival at the feedlot (-4%).There was no significant (p>0.05) effect of the BRSV vaccine on treatment rate in calves immunized at weaning, in calves immunized upon arrival at the Saskatoon bull test station, or in yearlings immunized upon arrival at the feedlot.Although the trend in these field trials was to a sparing effect of the BRSV vaccine, the small reduction in treatment rate may not justify the cost of the vaccination program. PMID- 17423525 TI - In defence of case reports. PMID- 17423526 TI - Follow-up on dogs licking humans' faces. PMID- 17423528 TI - Follow-up on dogs licking humans' faces. PMID- 17423527 TI - Criticism of Alberta preconditioned feeder review. PMID- 17423529 TI - Author's reply. PMID- 17423530 TI - Formation of the canadian association of zoo & wildlife veterinarians. PMID- 17423531 TI - VIDO sponsors day on BVD. PMID- 17423532 TI - A survey of brucellosis and tuberculosis in bison in and around Wood Buffalo National Park, Canada. AB - Examinations of complete or partial remains of 72 bison found dead in and around Wood Buffalo National Park, Canada, revealed evidence of brucellosis in 18 (25%) and tuberculosis in 15 (21%), with a combined prevalence of 42%. Urease-positive and ureasenegative strains of Brucella abortus biovar 1, and strains of biovar 2, were isolated from tissues of bison, including synovium and exudate from severe arthritic lesions. Mycobacterium bovis was isolated from a range of granulomatous lesions that were similar to those reported in tuberculous cattle. Diseased bison had a broad geographical distribution, and were found outside the park on at least three natural corridors. The diseases have a deleterious effect on this population of bison, and pose a health risk to other bison herds, livestock, and native hunters in the region. PMID- 17423533 TI - Current approaches to vaccine preparation. AB - Numerous conventional vaccines for animal use are currently available, and many of these vaccines have been instrumental in the control of infectious diseases of major economic importance. A vaccine has even been instrumental in global eradication of smallpox, an important human disease. However, many of the current vaccines are deficient in efficiency, potency, or safety. It has been recognized that the conventional methodologies are a limitation to further vaccine development. Introduction of monoclonal antibodies, recombinant DNA, and protein engineering techniques has facilitated a rather rapid increase in the knowledge of pathogenetic mechanisms, as well as of protective antigens at the molecular level. This knowledge provides the basis for development of a new generation of vaccines. As a rule, these vaccines contain purified immunogens, or even isolated epitopes, identified and prepared by molecular biological techniques. The efforts to find better delivery systems and better adjuvants accompany the research on vaccines. PMID- 17423534 TI - The usefulness of pelvic area measurements as an on-farm test for predicting calving difficulty in beef heifers. AB - Data collected from 197 replacement heifers in four beef cow herds in Saskatchewan were analyzed to determine the usefulness of pelvic area measurements as an on-farm test for predicting calving difficulty.The prevalence of calving difficulty in these herds ranged from 15-34%. Pelvic area measurements prior to the breeding season and at the time of pregnancy examination had poor positive predictive values and sensitivities in each herd. These findings were consistent whether the measurement used as a cut-off point to distinguish between a large and a small pelvic area was the lower 25th percentile, the mean, or the common standard. Standard pelvic area and calf birth weight ratios were not useful in estimating deliverable calf birth weight.Based on these results, pelvic area measurements prior to the breeding season and at the time of pregnancy examination are poor tests for predicting calving difficulty. PMID- 17423535 TI - A case-control study of Nocardia mastitis in Ontario dairy herds. AB - A case-control study was conducted to identify milking hygiene and udder therapy factors associated with a diagnosis of Nocardia mastitis in dairy herds from which milk samples are submitted to the Ontario Ministry of Agriculture and Food, Veterinary Laboratory Services, Guelph laboratory. The data were collected by telephone interview from 31 case and 31 control herds.After analytical control for confounders, blanket dry cow therapy and a dry cow antibiotic product were associated with a diagnosis of Nocardia mastitis in a herd, whereas another dry cow antibiotic product had a sparing effect.In comparison to mastitis pathogens susceptible to dry cow antibiotic therapy, Nocardia species were isolated infrequently from milk specimens submitted to the Guelph mastitis laboratory. PMID- 17423536 TI - Laminitis-like changes in the claws of feedlot cattle. AB - The purpose of this study was to observe and quantitate changes in the claws of two groups of feedlot cattle (calves and backgrounded yearlings) fed diets that varied in energy (73.5 or 78.5% TDN) and crude protein (11, 13, 15, 16, 17, or 19%) content. At slaughter, the thickness of sole horn and the prevalence of toe and heel hemorrhages were greater in calves than in yearlings (p<0.02). Feeding the high-energy ration increased the prevalence of toe and heel hemorrhages in calves (p<0.02) and heel hemorrhages in yearlings (p<0.02). In yearlings, rotation of the distal phalanx and ridging of the dorsal wall of the claw were the most prominent pathological features. Osteopathy of the apex of the distal phalanx occurred more frequently in calves than in yearlings (p<0.01). This study suggests that intensive feeding of beef cattle before they reach 14 months of age has a deleterious effect on digital health. PMID- 17423537 TI - Contagious ecthyma: An unusual distribution of lesions in goats. PMID- 17423538 TI - Use of epidural morphine to relieve pain in a horse. PMID- 17423540 TI - Compendium of animal rabies vaccines marketed in Canada. PMID- 17423539 TI - Cysts of a Toxoplasma-like organism in an Atlantic bottlenose dolphin. PMID- 17423541 TI - Saskatchewan. Salinomycin toxicity in turkeys. PMID- 17423542 TI - Prince Edward Island. Lyme disease vector, Ixodes dammini (the northern deer tick), identified in Prince Edward Island. PMID- 17423543 TI - Alberta. Death of a Rough Collie exposed to an ivermectin-based paste. PMID- 17423545 TI - Serum biochemical values in the skunk. PMID- 17423544 TI - British Columbia. Poisoning by lead shot in trumpeter swans. PMID- 17423546 TI - Veterinarians as behavior consultants. PMID- 17423547 TI - Les glucides et l'alimentation des chiens. PMID- 17423548 TI - Differences in life and disability insurance. PMID- 17423549 TI - Pet health insurance. PMID- 17423550 TI - Do we do enough genetic counselling? PMID- 17423551 TI - Modern beef production and disease control. PMID- 17423553 TI - Author's reply. PMID- 17423552 TI - More on the control of bovine spongiform encephalopathay. PMID- 17423554 TI - The impact of the GST on veterinary clinics: Taxing versus exemption versus zero rating. PMID- 17423555 TI - The genetic disease crisis in purebred dogs. PMID- 17423556 TI - Humane practices committee report. PMID- 17423557 TI - Serological titers to various leptospiral serovars before and after vaccinating gilts with three commercial vaccines. AB - The antibody response to various leptospiral serovars was evaluated in six-month old gilts vaccinated with three commercial vaccines, each containing five leptospiral serovars.All three vaccines elicited a significant postvaccinal antibody response to Leptospira canicola, grippotyphosa and icterohaemorrhagiae (copenhageni). The antibody response to the other leptospiral antigens in the vaccines varied among the vaccinates. None of the vaccinated groups developed significant titers to L. hardjo. Two of the three vaccines elicited a significant postvaccinal response to L. pomona, but in each case the titer mean of the vaccinated group was <1/100. Although not among the antigens in the vaccines, titers to L. bratislava increased in all vaccinated groups, except one, and in one control group. PMID- 17423558 TI - An evaluation of simultaneous GnRH and cloprostenol treatment of dairy cattle with cystic ovaries. AB - In a randomized double-blind clinical trial, 75 cows with ovarian cysts were treated with the synthetic gonadotropin releasing hormone, gonadorelin acetate (GnRH). Forty-two of these cows were simultaneously treated with cloprostenol (CP), and the remaining 33 cows received sterile saline. Milk progesterone (P(4)) was measured at treatment and two days later. Clinical response 30 days after treatment was determined by palpation per rectum, and estrus and breeding dates were recorded up to 90 days after treatment. Cows were examined for pregnancy by palpation per rectum 40 days or more after breeding. Milk progesterone levels two days after treatment were significantly lower and the 30-day clinical response rate was significantly higher in the GnRH + CP group than in the GnRH group. Intervals to first estrus and to conception, proportion in heat by day 21 after treatment, and pregnancy rate by 90 days did not differ significantly between the groups. The same relationships held in a subset of cows with P(4)>/=1 ng/mL at treatment. Fewer cows in the GnRH + CP group became pregnant by day 90 after treatment, but this difference was not significant. These results suggest that simultaneous GnRH and cloprostenol treatment of all cows with cystic ovaries cannot be recommended at this time. PMID- 17423559 TI - Prophylactic tilmicosin medication of feedlot calves at arrival. AB - The parenteral administration of the antibiotic tilmicosin given on arrival at a feedlot was evaluated in a group of 304 steer calves. These calves were allotted to 24 pens so that there were 12 replicates of both the control and medicated groups. The treatment rate was reduced significantly during the first five days (p < 0.05) and during the first month (p < 0.01) of the feeding period in the medicated group. The average days from arrival until first treatment for respiratory disease was increased to 21 days in the medicated group compared to 9 days (p < 0.01) for the controls. The medicated group had improved average daily gain (p < 0.01) and feed efficiency (p < 0.01) over the trial period when compared to the nonmedicated animals. PMID- 17423560 TI - Fractures of the femur in newborn calves. AB - A retrospective study of femoral fractures in 77 calves revealed that most occurred in the femoral shaft or in the proximal physis, with thin cortices and periosteal stripping significant problems in fractures of the femoral shaft. Most cases were related to forced extraction, with more left femurs being affected than right. The existence of concurrent disease reduced the success rate by 50% and, when significant additional trauma was present, no calves survived. When all cases were considered regardless of condition, the "longterm" success rate was 43%. PMID- 17423561 TI - Terminal ileitis in lambs. AB - Terminal ileitis was diagnosed in three flocks of lambs in different areas of Alberta. Salient clinical features in affected lambs were progressive emaciation with diarrhea, and in some lambs, frequent abdominal stretching. Postmortem findings included thickening of the ileal, and in some animals, the jejunal, cecal and colonic mucosa as a result of mucosal infiltrates of many lymphocytes and fewer plasma cells, eosinophils, globular leukocytes, and neutrophils. Hyperplasia of intestinal lymphoid tissue was prominent in most affected lambs. The cause of the condition is unknown. PMID- 17423562 TI - A rehabilitation team approach to correct flexural deformities in a donkey foal. PMID- 17423563 TI - Polioencephalomalacia in a captive fallow deer. PMID- 17423564 TI - Saskatchewan. Prevalence of Campylobacter jejuni in pronghorns and mule deer in southern Saskatchewan. PMID- 17423565 TI - Ontario. Satelliting streptococci. PMID- 17423566 TI - Alberta. Congential storage disease of Salers calves. PMID- 17423567 TI - British Columbia. An outbreak of aspergillosis in wild waterfowl. PMID- 17423568 TI - Meeting the need for veterinary diagnostic services in developing countries. PMID- 17423569 TI - Socio-economic importance of the diagnosis of disease in the backyard species in the Caribbean. PMID- 17423570 TI - Current Topics in nutrition. PMID- 17423571 TI - Schofield Centennial: Reminiscences of one of Dr. Schofield's students. PMID- 17423573 TI - Redefining our editorial policy. PMID- 17423574 TI - The addiction as disease theory. PMID- 17423575 TI - The theory of relativity and early pregnancy diagnosis examination in dairy cows. PMID- 17423576 TI - Authors' reply. PMID- 17423577 TI - Holistic veterinary medicine questions. PMID- 17423578 TI - Author's reply. PMID- 17423579 TI - Evaluation of colostrum supplements. PMID- 17423580 TI - Authors' reply. PMID- 17423581 TI - Authors' reply. PMID- 17423583 TI - Open letter to the Honorable Michael Wilson, Minister of Finance. PMID- 17423582 TI - Diagnosis of monensin intoxication in poultry. PMID- 17423584 TI - Food safety in a nuclear crisis: The role of the veterinarian. AB - Veterinarians are integrally involved in the maintenance of a safe and sufficient food supply; this involvement may be put to the test when a crisis - particularly a nuclear crisis - occurs. Few people can be said to be experts in how to ensure a safe food supply in a nuclear crisis. However, a working knowledge of radionuclides and how they behave in the food system, and the ability to identify high-risk foods and high-risk consumers in a crisis situation should be a part of the education of all veterinarians. In this paper I review those features of postdisaster radionuclide contamination of the food chain most relevant to veterinary activities in ensuring a safe food supply. PMID- 17423585 TI - Prevalence of Campylobacter jejuni in ranch mink at pelting: Cultural, serological, and histological evidence of infection. AB - This survey of 500 mink on three Wisconsin ranches at pelting gives an estimate of the prevalence of Campylobacter jejuni in the feces of clinically normal animals. On ranches 1 and 2, which used wet feed, C. jejuni was isolated by colon content culture from 7% and 32% of mink one year, and 43% and 13% the next year; the 200 bile samples tested were culture-negative. On ranch 3, which fed a pelleted ration, the organism was never isolated. Among culture-positive mink tested, 22 of 55 had bacterial agglutination serum titers to homologous and/or heterologous Campylobacter isolates from the ranch of origin. Four of 23 culture negative animals tested had titers. No histological evidence of inflammatory changes in the lower ileum and/or colon was found, although Campylobacter-like organisms were rarely seen in silver-stained sections from both culture-negative and culture-positive animals. We conclude that the presence of C. jejuni in the mink gut does not necessarily indicate a role in gastrointestinal disease. PMID- 17423587 TI - Endometrial biopsy in mares under nonuniform breeding management conditions: Prognostic value and relationship with age. AB - Endometrial biopsy plays an important role in fertility examination of the mare. The purpose of this study was to examine the prognostic usefulness of endometrial biopsy under nonuniform breeding management conditions, and to clarify any relationship with age of the mare. Based on endometrial histological changes, 192 mares from different farms were classified using a four-category grading system. There were significant associations between categories and fertility. Mean pregnancy rates in the four categories were 79, 49, 33 and 0% and mean foaling rates were 70, 42, 18 and 0%, respectively. Mares older than the median age (=11 years) had, on average, higher category scores and significantly lower fertility than younger mares. Within categories, there was a tendency towards higher fertility in younger mares. Although the precise prediction of fertility of the individual mare remains difficult, it is evident from this investigation that endometrial biopsy has considerable prognostic potential even under nonuniform breeding management conditions. Furthermore, the age of a mare must be considered when estimating her fertility potential. PMID- 17423586 TI - Campylobacter jejuni abortions in two beef cattle herds in Saskatchewan. AB - Abortions, accompanied by placental retention and weight loss, occurred during February and March in 19% of 120 and 10% of 108 beef cows and heifers on two neighboring ranches in southern Saskatchewan. A diagnosis of Campylobacter jejuni abortion was made based on lesions of necrotizing and suppurative placentitis and fetal bronchopneumonia in association with the culture of large numbers of C. jejuni from placentas and fetal tissues.Campylobacter jejuni was isolated with variable frequency from fecal samples of aborting and healthy cows, and scouring and healthy calves. Campylobacter jejuni serotype 2 (Lior) was isolated from fetal tissues and feces of a scouring calf, whereas C. jejuni serotypes 1, 4, 5 and 99 were isolated from feces of in-contact cattle. We hypothesized that the source and mode of transmission of C. jejuni was fecal contamination of water supplies and feeding grounds by carrier cows or wildlife. PMID- 17423588 TI - Roxarsone (3-nitro-4-hydroxyphenylarsonic acid) poisoning in pigs. AB - Young pigs, six to ten weeks of age, from two unrelated swine operations were fed a grower ration obtained from a common commercial supplier. Following ingestion of the feed for approximately two weeks, pigs in both groups developed neurological disturbances characterized by blindness, ataxia, incoordination, muscle tremors, posterior paralysis, and quadriplegia. Vocalization described as "screaming" was also observed in several animals. Necropsy findings and tissue arsenic concentrations were consistent with a diagnosis of phenylarsonic acid poisoning. The liver and kidney contained an average arsenic content of 2.9 and 1.8 mg/kg (wet weight), respectively. The feed contained 38 mg of arsenic/kg corresponding to 133 mg roxarsone (3-nitro-4-hydroxyphenylarsonic acid)/kg. This level of roxarsone is approximately three to five times higher than the levels recommended for swine rations. The feed company had placed roxarsone in the ration at levels recommended for the less toxic arsanilic acid. It was assumed that the two organic arsenicals could be added to the rations interchangeably at the same level of formulation. The present investigation indicated that roxarsone is more toxic than arsanilic acid and the margin of safety in swine rations is low. PMID- 17423589 TI - Ehlers-Danlos syndrome in a dog. PMID- 17423590 TI - Regurgitation due to megaesophagus in a ram. PMID- 17423591 TI - Ontario. Selenium toxicosis causing focal symmetrical poliomyelomalacia in pigs. PMID- 17423593 TI - Western Canada. Rabies diagnoses in Western Canada from July 1 - December 31, 1989. PMID- 17423592 TI - Quebec. Distribution of Streptococcus suis capsular types 9 to 22 according to the site of isolation. PMID- 17423594 TI - Western Canada. Botulism in wild waterfowl: Alberta, Manitoba and Saskatchewan. PMID- 17423595 TI - Survey results on CVMA insurance program. PMID- 17423597 TI - Critical writing and reading of review articles. PMID- 17423598 TI - Wildlife management. PMID- 17423599 TI - Animal rights/animal welfare. PMID- 17423600 TI - More on euthanize. PMID- 17423601 TI - Euthanize or euthanatize? PMID- 17423602 TI - Nocardial mastitis selective medium. PMID- 17423604 TI - Animal rights activists. PMID- 17423603 TI - Correction. PMID- 17423605 TI - An evaluation of vulvomucosal injections of prostaglandins for induction of parturition in swine. AB - Two trials were performed to evaluate the efficacy of prostaglandins administered via the vulvomucosal route at one-half the recommended dosage in comparison to prostaglandins injected intramuscularly (IM) at the standard dosage. In trial 1, sows on three commercial swine farms were given prostaglandin F2alpha at a dosage of 10 mg IM (n = 110) or 5 mg prostaglandin F2alpha using a vulvomucosal injection (n = 94). The numbers of sows farrowing within 36 h postinjection were 92 (84%) and 83 (88%), respectively. In trial 2, sows on four commercial swine operations were induced to farrow by means of one of three treatments: cloprostenol 175 mug IM (n = 71); cloprostenol 87.5 mug vulvomucosally (n = 57); or prostaglandin F2alpha 5 mg vulvomucosally (n = 96). The numbers of sows farrowing within 36 h postinduction were 69 (97%), 53 (93%), and 91 (94%), respectively.Vulvomucosal injections of prostaglandin F2alpha and cloprostenol at one-half the dosage appeared to be as effective as intramuscular injections of prostaglandin F2alpha and cloprostenol at the recommended level. There were fewer sows demonstrating restless behavior following the injection of lower dosages of prostaglandin F2alpha vulvomucosally, compared to sows given the recommended dosage of prostaglandin F2alpha IM. PMID- 17423606 TI - A case control study of fowl pox in southeastern Ontario. AB - An outbreak of fowl pox, which occurred in south-eastern Ontario between July 1988 and April 1989, was investigated in the spring of 1989 to determine factors associated with the spread of the disease. Clinical fowl pox was confirmed on five farms (cases). Twenty-seven farms, out of 35 egg producers with quota from Durham region to Northumberland county, provided information as controls. Bivariate analyses were performed on mail survey data using Fisher's exact test and odds ratios. Although the tests of hypotheses lacked statistical power because of the small number of case farms, and barns, a number of significant associations were found. At the farm level, fowl pox infection was associated with pullets purchased from a particular pullet grower. At the barn level, fowl pox infection was associated with pullets from a particular grower, mixing different groups of pullets, and a trend towards having birds early in the laying period, and higher numbers of birds placed. Fowl pox-infected barns had higher mortality and lower egg production postoutbreak. The results may indicate that the virus enters the laying barn at, or near, the time new birds are placed. Better communication among producers, catch-and-fill crews, and others associated with the egg industry, as well as more complete records of dates, sources, and persons involved with pullet placements, are recommended. PMID- 17423607 TI - Actinobacillus suis infection in pigs in southwestern Ontario. AB - Actinobacillus suis was isolated from tissues of 39 pigs, 2 porcine lungs, and 1 uterine swab submitted for diagnostic evaluation from 24 farms in southwestern Ontario between 1985 and 1988. These isolates represented a gradually increasing incidence of herd outbreaks caused by A. suis in southwestern Ontario. The outbreaks were typified by sudden death in suckling or recently weaned pigs; 87% of the affected pigs examined at the laboratory were between two and 28 days old. Petechial to ecchymotic hemorrhages in the thoracic and abdominal organs accompanied by serofibrinous exudates in both cavities were the most common gross lesions. The lesions were characterized histologically by bacterial thromboembolism and necrosis randomly scattered in thoracic and abdominal organs. Occasionally, bacterial thromboemboli were surrounded by centrifugally radiating, eosinophilic, club-like colonies. Diffuse necrohemorrhagic myocarditis that was more severe in the atria, and diffuse subacute meningoencephalitis, were less frequent but distinctive lesions. Multiple litters were affected in most herd outbreaks, and mortality often approached 50% in affected litters. Although the A. suis organism was susceptible to nearly every antibiotic against which it was tested, the suddenness of herd outbreaks precluded attempts at treatment. PMID- 17423608 TI - Necrohemorrhagic enterocolitis caused by Clostridium perfringens type C in a foal. PMID- 17423609 TI - Micronema deletrix in the kidney of a horse. PMID- 17423610 TI - Culturing of bulk tank milk for the presence of Nocardia spp. PMID- 17423611 TI - Catnip: Its uses and effects, past and present. PMID- 17423612 TI - Northwest Territories. Rabies in a polar bear. PMID- 17423614 TI - British Columbia. Bursal cryptosporidiosis in broiler chickens. PMID- 17423613 TI - Ontario. Mast cell tumor in a puppy. PMID- 17423615 TI - Alberta. Translocation 1/29 in Charolais cattle. PMID- 17423616 TI - Juvenile lymphoma in a Holstein heifer. PMID- 17423617 TI - Robert Robinson, 1836-1901: One of OVC's First Three. PMID- 17423619 TI - Are you environmentally friendly? PMID- 17423620 TI - Response to the open letter from Dr. Grenn to the Honorable Michael Wilson (Can Vet J 1990; 31: 342-343). PMID- 17423622 TI - More on face-licking. PMID- 17423621 TI - Dr. Grenn's reply. PMID- 17423623 TI - On the CVMA GST report. PMID- 17423624 TI - Animal welfare movement. PMID- 17423626 TI - Epidemiology and preventive medicine: A report on the CAVEPM 1989 meeting. PMID- 17423625 TI - Concerns about game ranching. PMID- 17423627 TI - Heartworm in dogs in Canada in 1989. AB - In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively. PMID- 17423628 TI - Environment, respiratory disease, and performance of pigs in three Saskatchewan grower-finisher barns. AB - A microcomputer-based environmental monitoring system was used to monitor temperature, humidity, and ventilation rate continuously in three commercial grower-finisher swine barns in Saskatchewan. During the monitoring period, a group of pigs in each barn was examined for growth rate, amount of lung affected with pneumonia, and degree of atrophic rhinitis. In addition, the total bacterial colony forming particle count within the airspace of each barn was measured once each week.Significant differences existed among barns for daily maximum and minimum temperature, relative humidity, ventilation rate, and average bacterial colony forming particle counts. There was no difference among farms in the average percentage of lung affected with pneumonia, average snout atrophy score, or growth rate of the test animals.On one farm, there was a significant positive correlation between snout score and percentage of lung affected with pneumonia. On another farm, there was a significant negative correlation between percentage of lung affected with pneumonia and growth rate. PMID- 17423629 TI - An abattoir survey of urinary bladder lesions in cattle. AB - Postmortem examination of 21,340 cattle at an Ontario abattoir over a one-year period revealed macroscopic lesions in urinary bladders of 2,296 animals (10.8%). Chronic cystitis, the most frequent finding, was observed in 2,209 cases (10.4%); uroliths were found in 994 (4.7%) of the cattle with chronic cystitis. Other types of urinary bladder lesions included acute cystitis, developmental abnormalities, hyperplasia, neoplasia, emphysema, and a parasitic cystitis. PMID- 17423630 TI - Efficacy of ivermectin oral liquid for horses. PMID- 17423631 TI - Acute gangrenous mastitis due to Clostridium perfringens type A and Escherichia coli in a cow. PMID- 17423632 TI - Branchial cyst in a cat. PMID- 17423633 TI - Quebec. Isolation of Streptococcus suis from cattle. PMID- 17423635 TI - Ontario. Possible sulfamethoxazole/trimethoprim-induced hepatic necrosis in a dog. PMID- 17423634 TI - Ontario. Osseous metaplasia of the renal pelvis in slaughter swine. PMID- 17423636 TI - Alberta. Monensin toxicity in swine. PMID- 17423639 TI - On building a better Canada. PMID- 17423637 TI - Ventral edema in horses testing positive for Potomac horse fever. PMID- 17423640 TI - A comment on salinomycin toxicity in turkeys. PMID- 17423642 TI - On the value of a case report. PMID- 17423641 TI - Further to the correspondence on holistic veterinary medicine. PMID- 17423643 TI - Q fever - a review. AB - Q or "query" fever is a zoonosis caused by the organism Coxiella burnetii. Cattle, sheep and goats are the most common reservoirs of this organism. The placenta of infected animals contains high numbers (up to 10(9)/g) of C. burnetii. Aerosols occur at the time of parturition and man becomes infected following inhalation of the microorganism. The spectrum of illness in man is wide and consists of acute and chronic forms. Acute Q fever is most often a self limited flu-like illness but may include pneumonia, hepatitis, or meningoencephalitis. Chronic Q fever almost always means endocarditis and rarely osteomyelitis. Chronic Q fever is not known to occur in animals other than man. An increased abortion and stillbirth rate are seen in infected domestic ungulates.Four provinces (Nova Scotia, New Brunswick, Ontario and Alberta) reported cases of Q fever in 1989.A vaccine for Q fever has recently been licensed in Australia. PMID- 17423644 TI - Cartilage healing: A review with emphasis on the equine model. AB - Articular cartilage is a remarkably resilient tissue capable of withstanding considerable stress and repeated loading. Since this tissue has no blood vessels, nerve elements, or lymphatics, it is not surprising that it has a limited capacity for repair when damaged. In the horse, cartilage damage occurs as an occupational hazard. Furthermore, developmental defects such as osteochondrosis can lead to osteochondritis dissecans. Resultant cartilage flaps, fissures, and poorly organized subchondral bone produce disruption of joint surfaces.Veterinarians are often called upon to intervene when damaged cartilage has healed incompletely. Basic understanding of the physiology and repair mechanisms of cartilage is paramount to successfully managing such injuries. This literature review gives a brief overview of recently published clinical and experimental studies on the healing of cartilage. The discussion centers on the equine model. PMID- 17423645 TI - A field trial to evaluate the efficacy of a commercial Pasteurella haemolytica bacterial extract in preventing bovine respiratory disease. AB - A double blind, random, controlled field trial was conducted to ascertain the efficacy of a Pasteurella haemolytica bacterial extract (Presponse, Langford Inc., Guelph, Ontario) in the prevention of bovine respiratory disease and/or its effects. Calves from 13 ranches (n = 1140 calves) were assigned to one of four groups, namely: vaccinated at the ranch three weeks prior to shipping to the feedlot; vaccinated only on arrival at the feedlot; vaccinated at both locations; or not vaccinated at either location. Four replicates of auction calves (n = 731) were also assigned to either receive or not receive the vaccine on arrival at the feedlot.The vaccine did not effect a change in morbidity rates or weight gain. Total mortality rates were increased significantly, and mortality rates from respiratory disease tended to be increased in ranch calves that were vaccinated with Presponse at the ranch. In auction calves, the relapse rates were significantly lower in vaccinated calves. There was a tendency towards a reduction of respiratory disease-related mortality, however there appeared to be no sparing against death from fibrinous pneumonia in auction calves. PMID- 17423646 TI - Boar culling in swine breeding herds in Minnesota. AB - This investigation was conducted to study boar culling patterns in swine breeding herds. Data were obtained from 84 swine breeding herds and included 440 boars that were culled. Each producer was involved for 12 consecutive months, and recorded every boar that was removed from the herd, the date, and the reason for culling. The annual culling rate for the 84 farms averaged 59.4% +/- 6.4 (SEM). The correlation coefficient between boar and sow culling was 0.52 (p <0.0001). Removal was the result of being overweight (47%), reproductive problems (18%), leg problems (12%), death (7%), and other diseases (4%). From the annual culling rate, the average breeding life of boars was estimated at 20 months. From this study, we concluded that the annual culling rate for boars in commercial herds was high and related to several different factors. PMID- 17423647 TI - Mixed papillary-polypoid cystitis in a heifer. PMID- 17423648 TI - Relationship of cerebrospinal fluid protein concentration determined by dye binding and urinary dipstick methodologies. PMID- 17423649 TI - Effect of ivermectin on weight gain of calves on pasture in Alberta. PMID- 17423650 TI - Atlantic Canada. Dermocystidium-like parasite in farmed Atlantic salmon. PMID- 17423651 TI - Alberta. Thymic hypoplasia in weak platinum fox pups. PMID- 17423652 TI - Saskatchewan. Outbreak of neonatal infectious bovine rhinotracheitis. PMID- 17423653 TI - British Columbia. Paratyphoid infection in English sparrows. PMID- 17423655 TI - Interviewing pre-vet students. PMID- 17423656 TI - Euphemisms for "euthanize". PMID- 17423657 TI - Border disease - a cause of terminal ileitis in lambs? PMID- 17423659 TI - Development and approval of new vaccines. PMID- 17423658 TI - The essential ingredients of successful strategic planning and its implementation. AB - The strategic planning process initiated by the Pew Foundation through the Pritchard Report became a major preoccupation of all of the 31 faculties of veterinary medicine in the United States and Canada. It is proposed that, in order to be successful in the strategic planning process, three determinants related to perception, to personality, and to psychosocial behavior are necessary ingredients. First, the professional program should evolve in a team-spirited milieu which can inspire future veterinarians to work in teams. Second, all channels of communication should be kept open. Third, passion and enthusiasm should be the prevailing and all-encompassing characteristics of faculty endeavors. Team-spirit, communication and enthusiasm are the essential ingredients which will be needed to confront the challenges facing the profession, to look towards the future with tremendous hope, and to embrace the 21st century with eagerness. PMID- 17423660 TI - Chronic copper toxicity in a dairy herd. AB - The addition of excessive copper to a commercially prepared dairy ration caused chronic copper toxicity in a dairy herd. A formulation error by a feed company resulted in copper levels of 800 to 1,000 mg/kg in the "as fed concentrate," amounting to about 400-500 mg copper/kg of the whole ration. Five animals died with typical signs of acute copper toxicity, including intravascular hemolysis and methemoglobinemia. A further 39 cows died on the farm from a combination of debilitation and secondary infectious causes, and 215 were sent to slaughter because of debilitation and poor milk production. The mortality of calves born to dams that had been fed the toxic concentrate was approximately 50%.We postulate that dairy cows, particularly pregnant cows, may be more susceptible to copper toxicity than other cattle, and suggest reexamination of the presently allowable maximum levels of copper supplementation of diets for dairy cattle. PMID- 17423661 TI - Sudden death in turkeys with perirenal hemorrhage: Field and laboratory findings. AB - Sudden death associated with perirenal hemorrhage, pulmonary congestion and edema, splenic enlargement and mottling, and hepatic congestion affected rapidly growing, apparently healthy male turkeys. Concurrent lesions of other diseases occurred in some birds. No causative agent was identified by bacteriological examination.The number of diagnoses of this condition has been increasing in recent years. Mortality usually occurs in males at 8-13 weeks of age. This condition was seen throughout Alberta and was not restricted to poults from any specific hatchery, or on any specific feed. Fast weight gain, continuous lighting programs, crowding, and hyperactivity are possible important etiological factors. Various treatments have been used with no significant effect on mortality from the condition. PMID- 17423662 TI - Ablation of a congenital neck mass in a foal. PMID- 17423663 TI - An ulcerative enteritis of racing pigeons. PMID- 17423664 TI - Unilateral stylohyoid disarticulation as a cause of dysphagia in a dog. PMID- 17423665 TI - Prince Edward Island. Accidental electrocution of pigs. PMID- 17423667 TI - Alberta. Deposition of visceral urates associated with oxalate nephrosis in turkeys. PMID- 17423666 TI - Ontario. Canine ehrlichiosis in Ontario. PMID- 17423668 TI - Ontario. Incidental neoplasia during experiments on scrapie in white Swiss mice. PMID- 17423670 TI - Veterinary medical ethics. PMID- 17423671 TI - Zoonoses parasitaires d'origine canine transmises par lechage. PMID- 17423672 TI - Comments on game farming. PMID- 17423674 TI - Treatment of cystic ovaries. PMID- 17423673 TI - Author's reply. PMID- 17423675 TI - Assessing vaccine efficacy. PMID- 17423676 TI - An evaluation of antimicrobial therapy for undifferentiated bovine respiratory disease. AB - A field trial of antimicrobial therapy for cases of undifferentiated bovine respiratory disease (UBRD) in beef calves was conducted at four Ontario feedlots. The primary purpose of the trial was to evaluate the efficacy of three different antimicrobials (oxytetracycline, penicillin, and trimethoprim-sulfadoxine) in the treatment of UBRD occurring within the first 28 days postarrival.The response, relapse, and case fatality rates overall were 85.7%, 14.8%, and 1.4%, respectively, and were not significantly different among the three antimicrobials evaluated. Weight gains of calves treated with the different drugs were not statistically different over the feeding period. Calves that suffered a relapse posttreatment were first treated significantly earlier (p<0.001) in the postarrival period than those that did not relapse. Considered together, treated calves gained significantly less (p<0.05) over the first 28 days and throughout the entire feeding period than controls that were never sick. Cases of UBRD that responded to therapy and did not relapse had rates of gain that were not significantly different from the controls. PMID- 17423677 TI - Cesarean section in the sow: A retrospective analysis of litter size and stillbirth rate. AB - The purpose of this study was to examine the litter size and stillbirth rate associated with cesarean-derived litters and to examine the relationship between the number of previous cesarean sections a sow had experienced and litter size. The records of 856 cesarean sections during a ten-year period from 1978 to 1988 were examined. The mean litter size was 10.6 +/- 3.2 and the mean number of stillborn pigs per litter was 0.2 +/- 0.8. The low level of stillbirth observed in this study provides supportive evidence to many earlier publications suggesting that the majority of stillborn pigs die during the birth process itself.A negative correlation between the number of previous cesarean sections a sow had experienced and litter size (r(2) = 0.015, p < 0.001) was observed. The slope of the regression line was -0.55, suggesting that litter size is reduced by approximately one-half a piglet for each cesarean section the sow has previously experienced. There are many factors which influence the variation in litter size. The small correlation coefficient (r(2) = 0.015) observed in this study indicates that only 1.5% of the variation in litter size can be explained by the number of previous cesarean sections that the sow has experienced. PMID- 17423678 TI - Four bovine meningeal tumors. AB - Four bovine meningeal tumors from a total of 14 brain tumors were recorded in the files of the Ontario Veterinary College and the Ontario Veterinary Laboratory Services Branch from the years 1966 to 1984. The affected cattle ranged in age from 10 months to adult. They exhibited various nervous signs and, in each case, the animals became recumbent. Tumors were located in various areas of the central nervous system. Neoplastic cells had oval vesicular nuclei containing numerous cytoplasmic invaginations. Virus particles were not observed. A fibroblastic appearance, with the presence of loose whorls or bundles, was present in all. Three of the tumors were invasive and were classified as a fibroblastic meningioma. PMID- 17423679 TI - Antimesenteric enterotomy for repair of a dorsal rectal tear in a mare. PMID- 17423680 TI - The evaluation of an enzyme immunoassay for the assessment of progesterone in canine plasma. PMID- 17423681 TI - Alberta. Death of six young Labrador/Malamute dogs exposed to automobile exhaust. PMID- 17423682 TI - Quebec. Poultry litter associated botulism (type C) in cattle. PMID- 17423683 TI - Alberta. Pseudotuberculosis in a black-tailed ptarmigan. PMID- 17423684 TI - Exercise in electrocardiography. PMID- 17423685 TI - Walter George Boswell: Veterinary surgeon for the British North American Boundary Commission of 1872-76. PMID- 17423687 TI - Veterinary medical ethics. PMID- 17423686 TI - Compassion in veterinary medicine. PMID- 17423688 TI - Obtiuary: dr. Margaret maxwell. PMID- 17423689 TI - A comparison of three oral electrolyte solutions in the treatment of diarrheic calves. AB - Thirty-six diarrheic calves infected with rota- and coronaviruses were randomly allocated to one of three oral electrolyte treatments: Ion-Aid (Syntex Agribusiness), Life-Guard (Norden Inc), or Revibe (Langford Inc). The calves were also allowed voluntary access to milk which was offered at the rate of 5% of body weight per feeding in two feedings daily. There were significant differences in recovery rate among calves treated with the different electrolytes. Only 33% of Ion-Aid-treated calves recovered; Revibe- and Life-Guard-treated calves had high recovery rates of 92% and 83%, respectively. The much higher recovery rates with Life-Guard and Revibe were attributed to the presence of an alkalizing agent in these preparations. Life-Guard uses bicarbonate to counteract acidosis and there was some evidence that this may have interfered with milk digestion. Revibe uses acetate; this was effectively metabolized within the calves' tissues and produced alkalization without interference with milk digestion. PMID- 17423690 TI - Treatment of equine leg wounds using skin grafts: Thirty-five cases, 1975-1988. AB - A retrospective study was conducted on 35 equine patients with lower leg wounds that were managed utilizing skin graft procedures. Two pinch graft, five punch graft, seven tunnel graft, eight split-thickness mesh graft and thirteen full thickness mesh expansion graft procedures were performed in the initial treatment. The average wound size was 188 cm(2). Twentyfour cases had pregrafting complications: 10 wounds developed sequestra; three wounds were grossly contaminated and infected; and 11 cases developed granulation tissue complications prior to grafting. Graft failure following the initial procedure was seen in 12 cases and occurred with all techniques except pinch grafting. Graft failure was often attributable to poor quality of granulation tissue as well as anatomic site, especially the dorsal surface of the tarsus. An average of two additional grafting procedures was required to successfully treat initial failures. Pinch grafts took the longest time to epithelialize (70 days), followed by punch grafts (47 days). Both were similar in terms of being the least durable and least cosmetically acceptable of all techniques used. Split-thickness and full-thickness mesh expansion grafts were technically the most difficult, but showed the most rapid epithelialization (28 days), greatest durability, and the best cosmetic appearance. Tunnel grafts provided a practical technique for grafting cases which were either not suited for, or which had failed with, mesh expansion grafts. PMID- 17423691 TI - Nephrotoxicity in dogs associated with methoxyflurane anesthesia and flunixin meglumine analgesia. AB - Uremia unexpectedly developed in five dogs 24 hours after undergoing thoracotomy in a student laboratory. In all dogs general anesthesia had been maintained with methoxyflurane, muscle relaxation had been induced with gallamine, and each dog received a single intravenous dose of 1.0 mg/kg flunixin meglumine for analgesia upon termination of anesthesia. In a subsequent group of dogs undergoing an orthopedic procedure, we assessed the effects on renal function of methoxyflurane anesthesia plus oxymorphone, or of methoxyflurane or halothane anesthesia in combination with a single IM 1.0 mg/kg dose of flunixin meglumine. Significant elevations in serum urea and creatinine values, and necrosis of collecting ducts and loops of Henle, were noted only in the dogs receiving methoxyflurane and flunixin meglumine.We conclude that the use of combination of methoxyflurane and flunixin meglumine is contraindicated in dogs. PMID- 17423692 TI - A case of bovine mastitis caused by Listeria monocytogenes. PMID- 17423694 TI - Prince Edward Island. Isolation of Yersinia spp. from cases of diarrhea. PMID- 17423693 TI - Chemodectoma in a horse. PMID- 17423695 TI - Manitoba. Probable organophosphate toxicosis in commercial geese. PMID- 17423697 TI - Alberta. Infectious laryngotracheitis in a flock of layers. PMID- 17423696 TI - Alberta. Tracheal collapse/stenosis in calves. PMID- 17423698 TI - One family's contribution to veterinary medicine in North America. PMID- 17423699 TI - Feeding older dogs and cats. PMID- 17423701 TI - Thoughts on euthanasia. PMID- 17423702 TI - Bernese Mountain Dog tumor study. PMID- 17423703 TI - Veterinary ethics. PMID- 17423704 TI - Prevalence of antibodies to infectious bovine rhinotracheitis, parainfluenza 3, bovine respiratory syncytial, and bovine viral diarrhea viruses in cattle in Saskatchewan and Alberta. AB - A total of 1745 healthy cattle from 295 farms in Saskatchewan and Alberta was tested by ELISA for antibodies to four viruses. Antibodies to infectious bovine rhinotracheitis (IBR) virus were found in 37.8% of sera (59.5% of properties), to parainfluenza 3 (PI3) virus in 93.9% of sera (99.7% of properties), to bovine respiratory syncytial (BRS) virus in 78.5% of sera (86.6% of properties), and to bovine viral diarrhea (BVD) virus in 40.6% of sera (66.7% of properties)The prevalence of PI3 viral antibodies among Saskatchewan cattle was not affected by district of origin, breed, sex, age, or vaccination practices, though BRS viral antibodies appeared less frequent in young, male, and unvaccinated animals. Antibodies to IBR and BVD viruses were less prevalent in the Prince Albert/Tisdale districts and in young, male, and unvaccinated animals, but were more common in Holstein cattle. Antibodies to IBR virus appeared less frequent in Herefords. Antibodies were more prevalent in cattle which had been vaccinated against IBR, BRS, and BVD virus infections.The relatively small number of cattle sampled from Alberta had a similar prevalence of antibodies to PI3 and BRS viruses to that seen in cattle in Saskatchewan, though IBR and BVD prevalence rates were lower. PMID- 17423705 TI - Epidemiological features of calf mortality due to hemophilosis in a large feedlot. AB - Hemophilosis was the most significant cause of mortality in fall-placed calves in a large feedlot in Saskatchewan, despite routine single immunization of calves with a commercial Haemophilus somnus bacterin on arrival. Common manifestations of fatal H. somnus infection were myocarditis and pleuritis; occasionally, thrombotic meningoencephalomyelitis (TME), peracute septicemia, and pneumonia were observed. Circumstantial evidence suggested that H. somnus may be a pathogen in polyarthritis. Death from pneumonia mainly occurred during the first five weeks in the feedlot. Death from myocarditis, pleuritis, TME, and septicemia, and euthanasia because of polyarthritis, occurred mainly after the third week in the feedlot.The median fatal disease onset (FDO) for pneumonia was day 12; for septicemia, day 17; for polyarthritis, day 18; for myocarditis and pleuritis, day 22; and for TME, day 29. Calves that died from myocarditis frequently were found dead in their "home" pen; however, 88% of these animals had been treated previously. Fifty-seven percent of the calves that died from pleuritis were never treated, and those that died from TME or septicemia were either never treated or died shortly after initial treatment. PMID- 17423706 TI - Tilmicosin as a single injection treatment for respiratory disease of feedlot cattle. AB - Tilmicosin, a new semi-synthetic macrolide antibiotic, was evaluated in eight field trials as a single subcutaneous injection at dosages of 0 (placebo), 5, 10 and 20 mg/kg for the treatment of naturally occurring respiratory disease in feedlot cattle. Animals for these trials were selected from large groups of recently-shipped feeder cattle at the time clinical signs of respiratory disease and body temperature of 40.6 degrees C or higher were observed. Treated animals were evaluated daily for 10 days and finally at day 28. Each animal was weighed on the first day and again on day 28. Animals that died were necropsied. All treatment dosages were effective in significantly lowering mortality, improving weight gains, lowering body temperature, and reducing the severity of clinical signs when compared to the placebo-treated controls. Body temperature was the only variable with statistically significant differences among the dose levels. PMID- 17423707 TI - A survey of canine and feline skin disorders seen in a university practice: Small Animal Clinic, University of Montreal, Saint-Hyacinthe, Quebec (1987-1988). AB - Dermatological disorders accounted for 18.8% and 15.2%, respectively, of all the dogs and cats examined at the Small Animal Clinic, University of Montreal, Saint Hyacinthe, during a one-year period. In dogs, the most common groups of dermatological disorders encountered were bacterial folliculitis and furunculosis, allergic dermatitis, endocrinopathy, neoplasia, ectoparasitism, and immune-mediated dermatitis. The most common primary final diagnoses were bacterial folliculitis and furunculosis, atopy, food hypersensitivity, flea bite hypersensitivity, hyperadrenocorticism, and hypothyroidism. Breed predispositions were found for several canine dermatoses: bacterial folliculitis and furunculosis (collie, German shepherd, golden retriever, Newfoundland), atopy (boxer, golden retriever), food hypersensitivity (boxer, German shepherd), hyperadrenocorticism (miniature poodle), hypothyroidism (Doberman pinscher, Gordon setter), castration responsive alopecia (chow chow), demodicosis (Old English sheepdog), and idiopathic pruritus (pit bull terrier).In cats, the most common dermatoses were abscesses, otodectic mange, cheyletiellosis, flea bite hypersensitivity, atopy, flea infestation, neoplasia, and food hypersensitivity. Himalayan and Persian cats accounted for 50% of the cases of cheyletiellosis and 75% of the cases of dermatophytosis, respectively. Hereditary primary seborrhea oleosa was seen only in Persian cats. PMID- 17423708 TI - Severe proliferative and necrotizing pneumonia in pigs: A newly recognized disease. PMID- 17423709 TI - Physaloptera stomach worms associated with chronic vomition in a dog in Western Canada. PMID- 17423710 TI - Comparison of three anthelmintics in the control of intestinal nematodes in young horses on fall and winter pasture. PMID- 17423711 TI - Ontario. Suspected hypersensitivity to insect bites in market pigs. PMID- 17423712 TI - Ontario. Diseases in baitfish in Ontario. PMID- 17423713 TI - Alberta. Fowl cholera in overwintering mallard ducks. PMID- 17423714 TI - Isoflurane as an inhalational anesthetic agent in clinical practice. AB - Isoflurane is the most recently available inhalational anesthetic agent on the market today. Although there have been few clinical trials comparing its use to halothane and methoxyflurane, the pharmacology of the agent suggests certain situations in which it may be the preferable agent. These include avian anesthesia, geriatric patients, patients with cardiovascular disease or hepatic disease, critically ill and unstable patients, cases such as brachiocephalics where upper airway obstruction is a concern during recovery, patients where increases in intracranial pressure should be avoided, and cesarean section. In addition, the rapid recoveries seen with isoflurane may be an advantage for outpatient surgeries. PMID- 17423716 TI - Tail tip necrosis in Ontario beef feedlot cattle. AB - Studies were performed to establish the prevalence and importance of tail tip necrosis in the southern Ontario beef feedlot industry and to characterize the gross appearance and histopathology of the condition. In a mail survey, 96% of 71 feedlots with slatted floors, but only 5% of 184 feedlots with solid floors, reported a problem with tail tip necrosis from 1982-1986. Treatments reported included antibiotics, amputation of the tail (therapeutic or preventive), and slaughter. Lameness was associated with tail tip necrosis.A scoring system for severity of necrosis was developed. Repeated inspections revealed that mild lesions were unlikely to progress to more severe stages. Histological alterations such as perivascular edema and hemorrhage, dermal scarring, follicular atrophy, and paucity of leukocytes were compatible with cutaneous ischemia.Of 441 tails inspected at slaughter plants, 34.5% were affected, with 3.4% involving skin lacerations and infection, and 4.3% amputated before slaughter. PMID- 17423717 TI - Veterinarians in disaster planning. PMID- 17423718 TI - Analyte comparisons on the Coulter Dacos and Kodak Ektachem DT-60 clinical chemistry analyzers. AB - Eighteen analytes were compared using the Coulter Dacos and Kodak Ektachem DT-60 chemistry analyzers. All analytes were significantly linearly related. Correlations were excellent for fifteen analytes (r > 0.95); the correlations for sodium, chloride, and total protein were less than 0.95 but greater than 0.90. Several analytes had slopes significantly different from unity which was likely related to methodological differences. Regression equations are presented for the conversion of data generated using the Kodak Ektachem DT-60 to Coulter Dacos values so that, in the University of Guelph clinical pathology laboratory, one set of reference intervals serves both instruments. These data are directly applicable only in the laboratory in which they were developed. They should not be utilized directly in other laboratories, but can be used as general guidelines until confirmatory studies are done. PMID- 17423719 TI - Vaginal hyperplasia in the bitch: Literature review and commentary. AB - We reviewed the incidence, etiology, clinical presentations, diagnosis, treatments and prevention of canine vaginal hyperplasia, and describe a simple surgical technique for its correction as practiced at the Small Animal Clinic, State University of Utrecht, The Netherlands. Many different treatments have been used, and some are novel approaches. In many cases, prevention is accomplished if the bitch is ovariectomized; there is a high incidence of recurrence with medical treatment. PMID- 17423720 TI - An unusual case of canine vaginal hyperplasia. PMID- 17423721 TI - Veterinary medical ethics. PMID- 17423722 TI - Maxillary fibrous dysplasia in a yearling filly. PMID- 17423723 TI - Prince Edward Island. Meconium aspiration syndrome in neonatal calves. PMID- 17423724 TI - Ontario. Brucella ovis, une cause d'avortement chez le mouton. PMID- 17423725 TI - Western Canada. Western Canada rabies diagnoses January 1 - June 30, 1990. PMID- 17423726 TI - Saskatchewan. Listeriosis in rabbits. PMID- 17423727 TI - Introducing an employee incentive plan into your practice. PMID- 17423730 TI - CVMA 1991: The who, what, where, when and whys. PMID- 17423728 TI - Exercise in electrocardiography. PMID- 17423731 TI - Comparison of ivermectin, oxibendazole, and pyrantel pamoate in suppressing fecal egg output in horses. AB - Thirty resident horses at a boarding stable in Alberta were used to evaluate the relative efficacies of ivermectin, oxibendazole, and pyrantel pamoate in reducing fecal egg output in adult horses under routine management conditions during spring and early summer, and to more clearly define the duration of suppression of fecal egg production following anthelmintic treatment. Horses were blocked according to pretreatment egg counts and randomly assigned to one of three treatments: pyrantel pamoate at 6.6 mg/kg body weight; oxibendazole at 10 mg/kg body weight; or ivermectin at 200 mug/kg body weight. All treatments were administered orally as a paste on day 0.Fecal samples were collected for examination by the modified Wisconsin procedure before treatment, and then at 4 11 day intervals up to day 72.Very few if any strongyle eggs were found in the feces of any horses up to day 35. On days 42, 50 and 57, the geometric mean egg count for the ivermectin group was significantly (p<0.05) lower than that for the oxibendazole or pyrantel pamoate groups. Based on a survival curve analysis of the data, the mean number of days for recurrence of eggs in the feces was significantly longer for the ivermectin group than for the oxibendazole and pyrantel pamoate groups.Under conditions encountered in this study, the posttreatment interval to resumption of fecal egg out-put in horses treated with ivermectin was eight to nine weeks, compared with five to six weeks for horses treated with oxibendazole or pyrantel pamoate. PMID- 17423732 TI - Lead poisoning in bald eagles in British Columbia. PMID- 17423733 TI - Enilconazole as an adjunct to the treatment of four cases of canine nasal aspergillosis. PMID- 17423734 TI - Effect of pre-partum parenteral supplementation of pregnant beef cows with selenium/vitamin E on cow and calf plasma selenium and productivity. PMID- 17423735 TI - Canada. Update on Nocardia sp. mastitis. PMID- 17423736 TI - Ontario. Meningoencephalitis associated with bacterial kidney disease in salmonids. PMID- 17423737 TI - Alberta. Salinomycin toxicity in turkeys. PMID- 17423738 TI - Exercise in electrocardiography. PMID- 17423740 TI - Concern about leadshot for waterfowl hunting. PMID- 17423741 TI - Veterinary medical ethics. PMID- 17423742 TI - Is game farming really all that bad? PMID- 17423743 TI - ACV 1991, Tout Ce que vous voules savoir. PMID- 17423744 TI - The control of infectious diseases of the respiratory and digestive tracts of cattle. PMID- 17423745 TI - Follicular dynamics and superovulation in cattle. PMID- 17423746 TI - Production of multiple genetically identical farm animals by nuclear transplantation. PMID- 17423747 TI - Embryo-mediated pregnancy failure in cattle. PMID- 17423748 TI - Seeking a new editor-in-chief. PMID- 17423749 TI - Our contributions to animal welfare. PMID- 17423751 TI - Veterinary medical ethics. PMID- 17423750 TI - Economics of practice. PMID- 17423752 TI - Students of the CVMA build for the future. PMID- 17423753 TI - The role of management and the use of vaccines in the control of acute undifferentiated diarrhea of newborn calves. PMID- 17423754 TI - Efficacy of parenteral antibiotics for disease prophylaxis in feedlot calves. AB - Trimethoprim-sulfadoxine (TMPSDX) and two formulations of oxytetracycline (OTC) were examined for their prophylactic efficacy in feedlot calves when given by intramuscular injection on arrival at a large commercial feedlot. The study included 2,112 high-risk feeder calves that developed disease early in the feeding period. Both formulations of OTC reduced bovine respiratory disease morbidity during the first two weeks on feed and for the entire feeding period by 15-19% (p<0.05), and they also reduced all fatal fibrinous pneumonia by 67% and 84% (p<0.05). All three drugs significantly reduced all fatal disease in animals first treated during the second week on feed, but not for the overall feeding period. Oxytetracycline with 2-pyrrolidone reduced the incidence of all fatal disease by 44% (p<0.05) during the entire feeding period. The case fatality risk for calves first treated during the second week on feed was lower (p<0.05) in the TMPSDX group and in the OTC with polyvinyl-pyrrolidone group. PMID- 17423755 TI - Contribution of delta bilirubin to the interpretation of hyperbilirubinemia in the horse - a pilot study. AB - A study was conducted to examine the relationship of delta bilirubin to traditional bilirubin fractions to determine if delta bilirubin might assist in differentiating causes of hyperbilirubinemia in the horse. A recently introduced thin-film method was used for delta bilirubin determination. Thin-film and the traditional diazo methods were used for determining total, unconjugated, and conjugated bilirubin fractions. Sera from 38 healthy and 85 sick horses were examined.Thin-film serum delta bilirubin determination does not appear to assist in differentiating causes of unconjugated hyperbilirubinemia in horses. There was good correlation between thin-film and diazo-determined total bilirubin and bilirubin fractions. Only small changes in reference limits would appear to be necessary for clinical application of the thin-film bilirubin methods used in this study. PMID- 17423756 TI - Stability of factor VIII and von Willebrand factor in canine blood samples during storage. PMID- 17423757 TI - Abdominal mesothelioma in a dog. PMID- 17423758 TI - Alberta. Toxicosis caused by ammoniated hay in calves. PMID- 17423759 TI - Prince Edward Island. Symmetrical truncal alopecia in rats. PMID- 17423760 TI - British Columbia. Cryptosporidiosis in weanling piglets with enteritis. PMID- 17423762 TI - Vitamin C in the prevention of neonatal calf diarrhea. PMID- 17423761 TI - 1. When and why should I do an electrocardiogram? PMID- 17423763 TI - Sexism in the CVMA? PMID- 17423764 TI - Veterinarians and game farming. PMID- 17423766 TI - Clinical management of bovine respiratory disease. PMID- 17423765 TI - Veterinary medical ethics. PMID- 17423767 TI - A review of insecticide poisonings among domestic livestock in southern Ontario, Canada, 1982-1989. AB - From 1982 to 1989, inclusive, 20 poisonings were investigated by the Ontario Ministry of Agriculture and Food following ingestion by domestic livestock of granular insecticides including terbufos (13 poisonings), disulfoton (two poisonings), fonofos (two poisonings), phorate (two poisonings), and carbofuran (one poisoning); all are used for rootworm (Diabrotica spp.) control in corn. A further three poisonings of livestock occurred following the ingestion of the foliar insecticide, endosulfan (two poisonings), and the seed protectant insecticides diazinon plus lindane (one poisoning). There were six poisoning cases as a result of excessive topical applications of the three insecticides coumpahos, fenthion, and lindane as dusts or sprays to control external parasites. Together, these events caused the deaths of 258 domestic animals of which 200 were cattle, 23 were swine, and 35 were sheep. Not all deaths are reported to the Ministry and the cases reported here may only represent 30-50% of the actual deaths over the period. Based on total populations of livestock, the percent losses were very small but they represent serious losses to individual growers. The economic loss is estimated at $160,000 over the eight years, or $20,000 per annum, and this does not include veterinary costs.Some of the poisoned animals died within as little as three to four hours of ingestion while others were sick but survived for several days. Lethal doses of insecticide were found in the rumen, abomasum, or stomach of dead animals. Signs typical of cholinesterase inhibition caused by organophosphorus poisoning were observed in most cases. Cholinesterase readings were found to be zero in dying animals. Necropsy findings were rarely more than pulmonary edema or myocardial hemorrhage. Where organochlorine insecticides were ingested, convulsions were the major manifestation.Contamination of feed was most often accidental, and chemical analysis was most helpful in identifying both potent and minor sources, thus facilitating cleanup procedures. PMID- 17423768 TI - An investigation of risk factors for nocardial mastitis in central Alberta dairy herds. AB - A case-control study was undertaken during the summer of 1989 in central Alberta dairy herds to identify independent predictors of nocardial mastitis. Thirty seven herds with nocardial mastitis were matched with control herds based on herd size, milk production, and enrolment in Alberta Dairy Herd Improvement Services. Control herds were considered free of nocardial mastitis based on negative cultures of four weekly bulk tank milk samples and one composite milk sample collected during the same period from each lactating cow in the herd. A detailed questionnaire on herd management was completed during farm visits. The use of blanket dry cow therapy was not found to be a risk factor for nocardial mastitis. Dry cow therapy with intramammary products containing neomycin and the use of multidose vials of dry cow medications were the only predisposing factors identified as being significantly associated with nocardial mastitis in central Alberta dairy herds. Use of neomycin as a dry cow therapy increased the odds of nocardial mastitis occurring in these dairy herds by 169 times. PMID- 17423769 TI - Prognostic features and clinical presentation of acute idiopathic enterocolitis in horses. AB - Clinical and hematological changes observed on presentation of 47 horses referred to the Ontario Veterinary College with acute idiopathic colitis were analyzed for their prognostic features. Cases of acute enterocolitis were characterized by fever, dehydration, abnormalities of serum electrolyte concentrations, azotemia, hypoalbuminemia, and increased serum concentrations of muscle enzymes. Severely dehydrated horses were seven times more likely to die or be euthanized than those that were not dehydrated. Other factors associated with failure to survive included the following: increased hematocrit, increased number of band neutrophils, increased serum creatinine and urea concentrations, and decreased blood pH and increasingly negative base excess. The results of multivariate variable analysis (stepwise logistic regression) suggested that, among the variables tested, base excess was the best predictor of death or survival. Twenty of 47 horses died or were euthanized. Reasons for death or euthanasia included: severe disseminated intravascular coagulation, unresponsiveness of severe metabolic acidosis and hypoproteinemia to treatments, and severity of colonic lesions on exploratory laparotomy. Of the surviving horses, three developed chronic laminitis (two were destroyed) and five developed jugular vein thrombosis. Fourteen of 16 horses for which subsequent histories were available returned to normal function.Early recognition of the disease, combined with early and aggressive correction of dehydration and of acid-base imbalance, may be important determinants of survival in horses with acute idiopathic colitis. PMID- 17423770 TI - Comparison of serum osteocalcin concentration and alkaline phosphatase activity as indicators of bone activity in a foal following surgery. PMID- 17423771 TI - The causes of sow mortality: A retrospective study. PMID- 17423773 TI - Ontario. Histomoniasis in a rhea. PMID- 17423772 TI - Western Canada. Rabies diagnoses July 1-December 31, 1990. PMID- 17423774 TI - Saskatchewan. Abortion due to Coxiella burnetii (Q fever) in a goat. PMID- 17423775 TI - Alberta. Suspected ivermectin toxicity in kittens. PMID- 17423776 TI - Job performance appraisals - A powerful management tool. PMID- 17423778 TI - Getting the most out of your diagnostic lab. PMID- 17423777 TI - Diagnostic ophthalmology. PMID- 17423779 TI - Standards of practice. PMID- 17423780 TI - Idiopathic trigeminal neuritis in a dog. PMID- 17423781 TI - Veterinary medical ethics. PMID- 17423782 TI - The CVMA Humane Practices/Animal Welfare Committee: A review and steps forward. PMID- 17423783 TI - Accuracy of rectal palpation and of a rapid milk progesterone enzyme-immunoassay for determining the presence of a functional corpus luteum in subestrous dairy cows. AB - The accuracy of rectal palpation and of a rapid milk progesterone enzymeimmunoassay (EIA) for determining the presence of a functional corpus luteum (CL) in subestrous dairy cows was investigated, using the results of a radioimmunoassay (RIA) for progesterone in skimmed milk as the "gold standard". The ovaries and uterus of each of 359 subestrous cows from 32 dairy herds were palpated per rectum, the presence or absence of a functional CL was predicted, and a milk sample was collected for analysis by both the rapid EIA and the RIA.The nine clinicians participating in the study had a combined sensitivity of 82.6% and specificity of 52.6%, compared to the EIA which had a sensitivity of 90.3% and a specificity of 84.2% for predicting the presence of functional luteal tissue.It was concluded that the technique of rectal palpation was inaccurate at assessing the functional (progesterone-secreting) status of ovarian structures. In addition, a qualitative EIA was as sensitive and more specific than rectal palpation in predicting the presence of a functional CL in the subestrous cow. PMID- 17423784 TI - Aerobic bacterial isolates in horses in a university hospital, 1986-1988. AB - Bacterial isolations were reviewed from equine trachea, guttural pouch, uterus, wounds, abscesses, blood, synovial fluid, and abdominal fluid submitted to the Clinical Bacteriology Laboratory of the School of Veterinary Medicine at the University of Montreal for aerobic bacterial culture from 1986 to 1988. Of the 733 samples submitted, 324 (44%) were positive for bacterial growth, and 233 antimicrobial sensitivity tests were performed. Seventy-six percent of all positive samples yielded one bacterial species and two were isolated from 22% of positive samples. Streptococcus zooepidemicus, Escherichia coli, and Actinobacillus spp. were isolated from 39%, 18%, and 15% of the samples, respectively.Bacterial growth was most common from guttural pouches, wounds and abscesses, and transtracheal washes (TTW), but was less common from uterus, blood, abdominal fluid, and synovial fluids. Streptococcus zooepidemicus was the most common bacterium recovered from guttural pouches, TTW, uterus, and wounds and abscesses. Escherichia coli predominated in abdominal fluids, blood, and synovia. Bacterial sensitivities to common antimicrobials are presented. PMID- 17423785 TI - Enzyme immunohistochemical staining of formalin-fixed tissues for diagnosis in veterinary pathology. AB - Disease diagnosis often relies on the detection of specific antigens in tissue specimens. Enzyme-based immunohistochemical stains of formalin-fixed, paraffin embedded tissues may be used to identify antigens associated with viral, bacterial and protozoal microorganisms, autoimmunity, and neoplasia. The detection of antigens in routinely fixed tissues offers several advantages over other diagnostic techniques. Sample submission is convenient and facilitates safe handling of potential human pathogens. Retrospective studies of stored specimens are possible. The technique is relatively rapid and enables detection of nonviable microorganisms. In addition, the ability to detect antigens in fixed specimens allows simultaneous visualization of the antigen and the histological lesion which may enhance the accuracy of diagnosis. PMID- 17423786 TI - Patent ductus arteriosus in a Holstein calf: A two-dimensional and Doppler echocardiographic study of the ductus arteriosus and validation. PMID- 17423787 TI - Concurrent ehrlichiosis and babesiosis in a dog. PMID- 17423788 TI - Metabolic acidosis without dehydration in seven goat kids. PMID- 17423790 TI - Atlantic Canada. Cold water winter lesions in Atlantic Salmon. PMID- 17423789 TI - Quebec. Isolation of Staphylococcus felis from cases of external otitis in cats. PMID- 17423791 TI - Ontario. Proliferative and necrotizing pneumonia (PNP) of swine: the Ontario situation. PMID- 17423792 TI - Injectable anesthetic agents for cats. PMID- 17423793 TI - Changing the editorial guard. PMID- 17423794 TI - Is game farming really all that bad? - follow-up. PMID- 17423796 TI - Author's reply. PMID- 17423795 TI - Diagnosis of cryptosporidiosis in piglets. PMID- 17423797 TI - Hyperadrenocorticism in a cat. PMID- 17423799 TI - A look at the pet food certification committee. PMID- 17423798 TI - Veterinary medical ethics. PMID- 17423800 TI - Epidemiology and preventive medicine: A report of the CAVEPM/ACEMPV 1990 meeting. PMID- 17423801 TI - A benefit cost analysis of dry-cow mastitis therapy in Ontario dairy herds. AB - Data collected from 297 dairy farms in Ontario were analyzed to investigate the economic consequences of using dry-cow antibiotic therapy, and to demonstrate the elements of an economic evaluation. Benefit/cost ratios ranged from 0.5 to 31.0 depending on the methods used to assess the benefits of therapy. In general, within the assumptions outlined in this analysis, dry-cow therapy was found to be economically advantageous. However, many factors can influence milk production and somatic cell counts. In this observational study, it is possible that some such factors were confounded with the use of dry-cow therapy, and may have biased the estimates of economic impact. PMID- 17423802 TI - Staphylococcal adenitis in ranch mink in Ontario. AB - Staphylococcus intermedius was identified as the cause of an outbreak of acute adenitis of the cervical apocrine glands of neonatal mink, and the cause of vaginitis and mastitis in adults. The disease occurred with greater frequency in mutation mink with Chediak-Higashi syndrome than in pastel and dark color phases. PMID- 17423804 TI - Acute hepatic failure (Theiler's disease) in a horse. PMID- 17423803 TI - Myelofibrosis: Review of clinical and pathological features in fourteen dogs. AB - A clinicopathological study was performed on 14 dogs with myelofibrosis (MF), in order to correlate clinical, laboratory, and histomorphological parameters and investigate factors of prognostic significance. The clinical signs included fatigue, weight loss, anorexia, and diarrhea. Physical findings included pale mucous membranes and wasting/emaciation. The major laboratory observations were moderate to severe, poorly-responsive anemia with various degrees of marrow cellularity and fibrosis. All dogs with severe, non-responsive anemia should have a bone marrow core biopsy, stained for connective tissue, in order to detect myelofibrosis. Myelofibrosis regressed in six dogs. PMID- 17423805 TI - Ameloblastic odontoma in a calf. PMID- 17423806 TI - Restrictive cardiomyopathy in a cat with hypereosinophilic syndrome. PMID- 17423807 TI - Ontario. Another case of Japanese yew poisoning. PMID- 17423808 TI - Ontario. Abortions and stillbirths associated with generalized cytomegalovirus infection in a swine herd. PMID- 17423809 TI - Manitoba. Proliferative and necrotizing pneumonia of swine in Manitoba. PMID- 17423810 TI - Diagnostic ophthalmology. PMID- 17423811 TI - A hundred years of importation: The first animal quarantine station in North America; Levis, Quebec, 1876-1982. AB - Quarantine, as a means of preventing disease importation, has been used for people and animals since the mid-19th century in Canada. The first animal quarantine facility in North America was established at Levis, Quebec in 1876. This quarantine station existed at Levis until 1982 when it was closed and the function moved to Mirabel, Quebec, near the International Airport. Veterinarians were in charge during the life of the Levis Quarantine Station and some were also in charge of the Port of Quebec or a nearby District Office prior to the 1950's. In 1884 and 1886 the value of such a facility was illustrated in preventing the entry into Canada of contagious bovine pleuropneumonia and a vesicular disease. It was described in 1933 as "undoubtedly our most important quarantine station" and a year's operating costs as "trifling in comparison to losses which could occur if a foreign plague invaded this country". This facility's history also illustrated the close veterinary and human medical cooperation during the early days of organized veterinary medicine in Canada. The station was an example for the establishment of other such facilities in North America. PMID- 17423812 TI - Let's work together. PMID- 17423813 TI - Familial nephropathy in miniature schnauzers. PMID- 17423814 TI - More on sexism. PMID- 17423816 TI - Le Comite des pratiques humanitaires/sur le bien-etre des animaux de l'ACV: recapitulation et avenir. PMID- 17423815 TI - Veterinary medical ethics. PMID- 17423817 TI - "Benchmark" - a large observational study of Ontario beef breeding herds: Study design and collection of data. AB - The methods used in a large field study which investigated the health and productivity of Ontario beef breeding herds are described. Beginning in the calving season of 1986, 180 breeding herds on 170 randomly sampled farms were followed for a two year period, using producer records and annual farm visits.The response (cooperation) rate among the eligible producers initially contacted was 70%. Approximately two-thirds of producers maintained individual animal records, primarily calving season records. Approximately 40% recorded disease occurrences.The advantages, disadvantages, and interpretation of "on-farm" observational studies are discussed. PMID- 17423818 TI - Measures of herd health and productivity in Ontario cow-calf herds. AB - A cohort of cows and heifers in 180 separate breeding herds from 170 randomly sampled farms was followed from the 1986 breeding season through to the weaning of their calves in 1987. Data were collected from farm records, survey information collected during farm visits, and provincial government weaning weight records."Kilograms of calf weaned per female-exposed-to-breeding" was calculated as a summary measure of herd productivity. The lowest 25% of cow-herds produced less than 160 kg of calf weaned per cow-exposed-to-breeding, while the highest 25% exceeded 205 kg.Overall calf crop was 78.1% for cows and 78.5% for heifers. The 25th, 50th, 75th, and 90th percentile estimates for rates, which were components of calf crop (e.g. calving rate), were estimated. The component rates that most influenced calf crop were culling rate for cows and stillbirth rate for heifers. PMID- 17423819 TI - Sporadic, severe bronchointerstitial pneumonia of foals. AB - Bronchointerstitial pneumonia was diagnosed postmortem in 19 foals in a 10 year retrospective study of submissions to a diagnostic center in Ontario. Mean age at death was 2.0 +/- 0.05 (SEM) mo (range five days to four months). Fourteen of 19 were aged from 1.5 to 2.5 mo. Clinically, the disease was generally characterized by sudden onset of fever and increasingly severe dyspnea which developed into respiratory distress before death. Mean length of illness was 7.0 +/- 0.33 days (range 1-21 days). The disease appeared to affect only individual foals on 19 different farms.At postmortem, lungs were typically diffusely red, wet, firm, and failed to collapse. The major lesion recognized histologically was epithelial necrosis of alveoli and terminal bronchioles. Alveolar lumens contained large epithelioid cells, which were probably macrophages, and multinucleated syncytial cells were present in 16 of the 19 lungs. Inflammatory cells were sparse. Intraalveolar fibrin was prominent in all lungs. Bacteriological examination revealed no significant pathogen in 12 animals, but Rhodococcus equi was isolated from seven foals, associated in two animals with extensive abscesses. Viruses were not recovered from eight foals examined.On the basis of the similarity and severity of lesions, the sporadic nature of the disease, and the similar age at onset which appears to coincide with declining maternally-derived immunoglobulins, we speculate that this disease may be the result of a viral infection. PMID- 17423820 TI - Serological studies of infectious bovine rhinotracheitis, parainfluenza 3, bovine viral diarrhea, and bovine respiratory syncytial viruses in calves following entry to a bull test station. PMID- 17423821 TI - Angiostrongylosis with disseminated larval infection associated with signs of ocular and nervous disease in an imported dog. PMID- 17423822 TI - Congenital coloboma in a IIama. PMID- 17423823 TI - Fatal hemorrhage associated with incarceration of small intestine by the epiploic foramen in three horses. PMID- 17423824 TI - Prince edward island. Scrapie in prince edward island. PMID- 17423825 TI - Ontario. Procaine penicillin toxicity in pigs. PMID- 17423826 TI - Saskatchewan. Haemophilus somnus myocarditis versus thrombotic meningoencephalitis in western Canada. PMID- 17423827 TI - Diagnostic ophthalmology. PMID- 17423828 TI - Hiring the right employee. PMID- 17423830 TI - Association of amphibian and reptilian veterinarians. PMID- 17423831 TI - Author's reply. PMID- 17423832 TI - Our philosophy concerning animal rights. PMID- 17423833 TI - CVMA/MSD Agvet Heartworm campaign. PMID- 17423834 TI - A reply from the Communications Committee. PMID- 17423835 TI - Ivermectin sur les chatons. PMID- 17423836 TI - A woman chairman responds. PMID- 17423838 TI - A veterinary tour of Eastern Europe - The curtain is rising! PMID- 17423837 TI - Veterinary medical ethics. PMID- 17423839 TI - A brief review of infectious and parasitic diseases of wapiti, with emphasis on western Canada and the northwestern United States. AB - In this paper I review diseases reported in both captive and free-ranging wapiti in western North America, with some reference to diseases in captive red deer in Great Britain, Europe, New Zealand, and eastern North America.With the exception of coronavirus in neonates, few viral agents are reported to cause serious disease losses in wapiti in North America at this time. Bacterial diseases of current significance include brucellosis (focus in Wyoming), clostridial diseases, coliform enteritis of neonates, pasteurellosis, and necrobacillosis. The endoparasites most likely to be seen causing lesions in wapiti of western North America are lungworm (Dictyocaulus viviparus), arterial worm (Elaeophora schneideri), and, possibly, liver fluke (Fascioloides magna). Ectoparasites of importance to wapiti are Psoroptes cervinus and Dermacentor albipictus. Nutritional diseases are not covered in this review. PMID- 17423840 TI - Uterine torsion in a cat. PMID- 17423841 TI - Is canine leptospirosis underdiagnosed in southern Ontario? A case report and serological survey. AB - An eight-year-old city-dwelling Cairn Terrier was presented to a veterinary hospital in acute renal failure with evidence of hepatic insufficiency. The dog was treated symptomatically over three days, during which time vomiting was largely controlled, but it became jaundiced as hepatic insufficiency worsened. Leptospira pomona was demonstrated in large numbers by immunofluorescent staining of urinary sediment. It was isolated and its identity confirmed as L. pomona genotype kennewicki. The source of the infection was thought to be raccoons.Sera from 474 blood samples submitted for diagnostic purposes to two clinical pathology laboratories in southern Ontario were examined with the microscopic agglutination test for antibodies to selected leptospiral serovars. Of the sera tested, 39.2% reacted at titers >/=1:100 with one or more serovars, the majority of all sera (26.2%) reacting at low titers to canicola or icterohaemorrhagiae, or both. These reactions likely resulted from vaccination. A smaller proportion reacted to other serovars tested: autumnalis (3.8%), bratislava (8.2%), grippotyphosa (1.9%), hardjo (3.0%), and pomona (3.2%). Among dogs reacting to these latter serovars (other than bratislava), many had broadly cross-reacting and relatively high titers. One dog with a titer of 1:800 to pomona had had a disease typical of leptospirosis two years previously. Three other dogs with high titers to autumnalis, bratislava, or mixed serovars had clinical histories compatible with leptospirosis.We suggest that leptospiral bacterins for dogs in Ontario be broadened to include at least serovars autumnalis and pomona. PMID- 17423842 TI - Rabies diagnosis in western Canada, 1985-1989. AB - The results of laboratory examination of 20460 specimens for the diagnosis of rabies by the standard fluorescent antibody and mouse inoculation tests over a five year period are presented. Specimens were received from British Columbia, Alberta, Saskatchewan, Manitoba, and the Yukon and Northwest Territories. Of those examined, 9.96% were positive. The main reservoirs of rabies were skunks, bats, and foxes. During this observation period a rabies epizootic occurred in skunks in Saskatchewan. When both tests were performed, the fluorescent antibody test was found to agree with the mouse inoculation test in over 99% of cases. PMID- 17423843 TI - Colonic intussesception in a Holstein calf. PMID- 17423844 TI - Acute pancreatitis and bone marrow suppression in a dog given azathioprine. PMID- 17423846 TI - British Columbia. "Hepatitis-splenomegaly" syndrome in commercial egg laying hens. PMID- 17423845 TI - Acute lymphocytic leukemia and pseudohyperkalemia in a Jacob ram. PMID- 17423847 TI - Ontario. Severe parasitic hepatitis in pigs. PMID- 17423848 TI - Alberta. Border disease virus isolated from twin lambs. PMID- 17423849 TI - British Columbia. Heartworms in dogs in British Columbia. PMID- 17423851 TI - Good communication: Why is it a problem? PMID- 17423850 TI - Exercise in electrocardiography. PMID- 17423852 TI - Public advertising of Rx drugs - Disagreement with CVMA position. PMID- 17423854 TI - The Chairman of CVMA's Practice Committee responds. PMID- 17423853 TI - A reply from CVMA's Public Health and Safety Committee. PMID- 17423856 TI - Veterinary medical ethics. PMID- 17423855 TI - Regulations changed for animal movement. PMID- 17423857 TI - Biologics Evaluation Laboratory update. PMID- 17423858 TI - Diagnosis of equine stifle joint disorders: three cases. AB - The clinical, radiographic, arthrographic, arthroscopic, and pathological findings of three horses with femorotibial joint injuries are presented. Overall diagnostic accuracy is improved when clinical signs, arthrography, and arthroscopy are combined. Treatment of these injuries remains limited. One horse was euthanized, and two were treated by stall rest after diagnosis. Of the two surviving horses, one became a successful breeding animal, and the other was unable to perform as a pleasure horse. PMID- 17423859 TI - Monitoring reproductive performance of small dairy herds in veterinary practice. AB - A descriptive field study involving 87 herds (3608 cows) in two veterinary practices was conducted to compute mean values for a panel of reproductive herd parameters. A method of monitoring herds and identifying those herds experiencing reproductive inefficiency is reported.When comparing the means of herd indices for both practices, only the means for the index "percent in heat by 60 days" were significantly different. Overall, 20 herds were found to have at least one herd index which was significantly different from the mean for all herds. Fourteen herds were found to have significant reproductive inefficiency. If the index "percent problem cows" had not been used, 29% of the herds with reproductive inefficiency would not have been indentified. Our study suggests that it is useful to compare reproductive indices among herds, practices, and regions using a veterinary office microcomputer. PMID- 17423860 TI - Idiopathic thrombocytopenic purpura in a Holstein bull. PMID- 17423861 TI - Vasculitis and necrosis of the mandibular salivary glands and chronic vomiting in a dog. PMID- 17423862 TI - Effects of intramuscular injections of folic acid in sows on subsequent litter size. PMID- 17423863 TI - Alberta. Suppurative orchitis associated with Haemophilus somnus in a calf. PMID- 17423864 TI - Ontario. Inonophore (maduramicin) toxicity in pigs. PMID- 17423866 TI - Good communication: How can it be achieved. PMID- 17423865 TI - Diagnostic ophthalmology. PMID- 17423867 TI - Proliferative and necrotizing pneumonia. PMID- 17423868 TI - There is no such drug or word as ;euthanol'. PMID- 17423869 TI - A veterinary prisoner of conscience. PMID- 17423870 TI - Cryptosporidiosis associated with piglet enteritis - follow-up. PMID- 17423871 TI - Veterinary medical ethics. PMID- 17423872 TI - Haemophilus somnus and reproductive disease in the cow: A review. AB - Haemophilus somnus causes inflammatory disease in the genital tract of cows as reported in several field surveys and experimental trials. This organism can also innocuously colonize the healthy genital mucosa of the cow, which indicates its dual relationship with the host, that of pathogen and commensal. Experimental data indicate embryocidal capability of this pathogen suggesting a possible role in early embryonic death. Haemophilus somnus also causes sporadic abortions after a bacteremia in the dam. Retrograde infection of the pregnant uterus from the lower genital tract appears unlikely; however, this process can account for post parturient endometritis. Detection of high homologous IgG(2) serum antibody titers using an ELISA test may be useful for the diagnosis of current or recent genital inflammation. Experimental laboratory data indicate that a proportion of genital strains of H. somnus are pathogenic and capable of causing thrombotic meningoencephalitis and perhaps pneumonia. In vivo testing of the pathogenicity of genital strains remains to be conducted. PMID- 17423873 TI - Hepatic disease in Alberta horses: A retrospective study of 'alsike clover poisoning' (1973-1988). AB - Over sixteen years, 49 horses were diagnosed by Alberta Agriculture Animal Health laboratories as having "alsike clover poisoning". There was a distinct northwestern distribution of cases, the majority coming from the Peace River district. This distribution is opposite to that of the Alberta horse population, but coincides with areas of alsike clover cultivation. Cases could be divided into chronic or nervous clinical presentations, as described by Schofield. Tissues from 45 animals were retrieved and examined microscopically. Significant histological lesions were confined to the liver and consisted of biliary fibrosis and epithelial proliferation.I conclude that alsike clover poisoning is a specific disease entity, likely due to exposure to an exogenous toxin. The evidence is not strong enough to incriminate alsike clover as the etiology. PMID- 17423874 TI - Management of chemical burns of the canine cornea. AB - Significant clinical signs and general principles of treatment for chemical burns of the canine cornea are presented using three typical case studies for illustration. Alkali burns are more common in dogs than acid burns. The sources of alkali in this study were soap, cement, and mortar dust. Common signs of chemical burns are ocular pain, corneal ulceration, tear film inadequacy, corneal edema, and marked corneal neovascularity. Successful treatment requires thorough ocular lavage, treatment for corneal ulceration, and adequate anti-inflammatory therapy when the corneal epithelium becomes intact. PMID- 17423875 TI - The relative economics of feeding open, aborted, pregnant feedlot heifers. AB - A 90-day finishing trial involving 144 feedlot heifers was conducted to compare the performance parameters and carcass characteristics of open heifers, therapeutically aborted heifers, and pregnant heifers. In the first 28 days of the trial, the aborted heifers had reduced (p < 0.05) feed intake (FI), average daily gain (ADG), and feed efficiency (FE) compared to pregnant and open heifers. Over the entire trial, on a live weight basis, the aborted group had reduced (p < 0.05) final weight, ADG, and FE compared to pregnant and open heifers. However, when the data were adjusted for total uterine weight, the aborted and open heifers had improved (p < 0.05) final weight, ADG, and FE compared to pregnant heifers. The aborted and open group had a higher (p < 0.05) carcass weight, rib eye area, dressing percentage, and cutability estimate compared to the pregnant heifers. The aborted group had lower (p < 0.05) carcass weight than the open heifers. Over the entire 90-day feeding period, there were no statistically significant differences among the groups with respect to feed intake (FI), average fat, grade fat, and carcass grades. Also, there were no significant health problems or mortality in any of the groups.In the economic analysis, aborted heifers returned $26.41 per head more than pregnant heifers. Open heifers returned $39.94 per head more than aborted heifers, and $66.35 more than pregnant heifers. Thus, aborting feedlot heifers during the second trimester was determined to be a safe and cost effective management decision. PMID- 17423876 TI - Separation anxiety. PMID- 17423877 TI - Postnatal cerebellar cortical degeneration in Labrador Retriever puppies. PMID- 17423878 TI - Porphyria in a New Zealand White rabbit. PMID- 17423880 TI - Ontario. Multiple organophosphate poisoning in a beef herd. PMID- 17423879 TI - Plasmacytic enterocolitis with hypoganglionosis in a puppy. PMID- 17423881 TI - Ontario. Cryptosporidiosis, rotaviral, and combined cryptosporidial and rotaviral infections in goat kids. PMID- 17423882 TI - Alberta. Herpesvirus-like viral infection in a rabbit. PMID- 17423883 TI - Saskatchewan. Diazinon toxicity in Canada geese. PMID- 17423884 TI - Managing demand and supply in your small animal practice. PMID- 17423886 TI - Flexibility in undergraduate veterinary educational programs. PMID- 17423887 TI - Live-calf fetotomy is unacceptable. PMID- 17423888 TI - Practitioners' dual obligation. PMID- 17423889 TI - Misdirected idealism. PMID- 17423890 TI - The heifer is the patient, not the calf. PMID- 17423891 TI - Inexpensive, effective rabies vaccinations for high risk occupations. PMID- 17423892 TI - Withholding times for Procaine Penicillin G in cattle. PMID- 17423893 TI - Veterinary medical ethics. PMID- 17423894 TI - The future of veterinary education in Canada: Highlights of the plenary session held during the CVMA Convention (July 1991). PMID- 17423895 TI - Quantifying the occurrence of early embryonic mortality on three equine breeding farms. AB - This prospective field study was designed to describe the incidence of early embryonic mortality (EEM) and factors associated with the cause of EEM on three equine breeding farms in Ontario during the 1989 breeding season. Early embryonic mortality was defined as the loss of a single embryo during the first 40 days of pregnancy (day 0 = day of ovulation or last breeding). Pregnancy diagnoses and subsequent embryonic losses were observed by serial trans-rectal ultrasonography between days 12-20 (PD1) and 21-30 (PD2), and by trans-rectal ultrasonography or palpation per rectum between days 31-40 (PD3). Information on pregnancy status of a mare (or cycle) at 40 days after the last breeding was recorded when available. Nonpregnancy rates were calculated on a per cycle basis, to account for mares with no ultrasonic evidence of an embryo at the initial pregnancy examination. Embryonic mortality rates per cycle were calculated cumulatively (EMR((40))) for the entire 40 day embryonic period and during the specific time periods when a pregnancy diagnosis took place (EMR((PD1)), EMR((PD2)), EMR((PD3))). Embryonic mortality rates were also calculated on a per mare basis for mares experiencing EEM on either their first (EMR((f))) or any (EMR((a))) breeding cycle. Per cycle mare withdrawal rates were calculated cumulatively for the entire 40 day embryonic period (MWR((40))), and at each specific pregnancy diagnosis time period (MWR((PD1)), MWR((PD2)), MWR((PD3))) to account for those breeding cycles in which mares were not able to be observed for the entire forty days of the embryonic period. Records from a total of 699 mares involving 1014 breeding cycles were examined and analyzed. Per cycle risk rates for nonpregnancy (NP) were 36.4%, 45.0%, and 22.1%, for farms 1,2 and 3, respectively. Per cycle EMR((40)) ranged from 8-17%. Per cycle MWR((40)) ranged from 56.5-98.9%, indicative of a high rate of mare withdrawal from the study for the duration of the "embryonic" period. Significant differences (p < 0.05) in EMR((f)) and EMR((a)) per mare existed between two farms, indicative of a farm effect on EMR. Multivariable forward stepwise logistic regression analyses revealed that mares bred on foal heat were 1.9 times more likely than mares not bred on foal-heat to experience EEM (p = 0.008). PMID- 17423896 TI - A case-control study of Nocardia mastitis in Nova Scotia dairy herds. AB - A case-control study was conducted to identify herd production, housing, and hygienic and therapeutic factors associated with a diagnosis of Nocardia mastitis in dairy herds in Nova Scotia. The data were collected by on-farm interviews with owners of 54 case and 54 control herds.Logistic regression was used to study risk factors. The use of dry cow products containing neomycin, including two specific dry cow products, was strongly associated with a diagnosis of Nocardia mastitis in a herd. Other factors which increased the risk of Nocardia mastitis were higher levels of production, larger herd size, and a large percentage of cows treated with dry cow products. These results are compared to results from a similar study carried out in Ontario. PMID- 17423897 TI - Isolation of border disease virus from twin lambs in Alberta. AB - We describe herein a field case of border disease (BD) in twin lambs. Both lambs were unthrifty, stunted, and one exhibited nervous signs characteristic of BD, with tremors of the head, neck, hind legs, and pelvis. Hairiness of the coat and excessive pigmentation, often seen in lambs with BD, were not observed. A noncytopathic virus, which showed cross-reactivity with bovine viral diarrhea (BVD) virus antiserum and BVD virus monoclonal antibodies, was isolated repeatedly from leukocytes from one lamb and from tissues of the other. Although the source of the virus is unknown, our results suggest that the dam of the affected twins had been infected during pregnancy. We used the BD virus isolated to inoculate pregnant ewes and experimentally reproduce the disease in a newborn lamb. Our findings indicate that leukocytes, rather than serum, should be utilized for BD virus isolation. Further, it is recommended that BD virus, rather than BVD virus, be used in serum neutralization tests when screening sheep for antibody titers. PMID- 17423898 TI - Anaerobic orbital cellulitis and septicemia in a dog. PMID- 17423899 TI - Isolates of Brucella suis biovar 4 from animals and humans in Canada, 1982-1990. PMID- 17423900 TI - Partial gastrointestinal obstruction for one month due to a linear foreign body in a cat. PMID- 17423901 TI - Western Canada. Rabies diagnosis in Western Canada, January 1 - June 30, 1991. PMID- 17423902 TI - Ontario. Disseminated histoplasmosis in a young dog. PMID- 17423903 TI - Alberta. Lowest ever bulk tank somatic cell count in the province. PMID- 17423904 TI - Theodore Augustus Girling (1876-1919): Canadian veterinarian and Great War poet. PMID- 17423906 TI - Prescription drug advertising. PMID- 17423907 TI - Veterinary specialization in Canada. PMID- 17423908 TI - CVMA Public Health and Safety Committee responds. PMID- 17423909 TI - CVMA Practice Committee Chair responds. PMID- 17423910 TI - Further information on diseases of wapiti. PMID- 17423911 TI - Veterinary medical ethics. PMID- 17423912 TI - Guidelines for euthanasia of domestic animals by firearms. AB - All animals that are to be killed, whether for food, for humane reasons, or because they are homeless, must receive a quick and painless death. In some smaller communities, veterinary or humane society expertise may not be readily available to humanely kill stray and unwanted animals. An alternative that provides for a humane death for the animal is by shooting. The following guidelines are intended to assist persons who must perform this usually distasteful task; they contain recommended techniques that will help to ensure that any animals killed by shooting will die in a humane way. PMID- 17423913 TI - Comparative serological response in calves to eight commercial vaccines against infectious bovine rhinotracheitis, parainfluenza-3, bovine respiratory syncytial, and bovine viral diarrhea viruses. AB - A field trial was conducted to compare the serological responses in calves to eight commercial vaccines against infectious bovine rhinotracheitis virus (IBRV), parainfluenza-3 virus (PI3V), bovine respiratory syncytial virus (BRSV), and/or bovine viral diarrhea virus (BVDV). Calves given IBRV, P13V, BRSV, and BVDV vaccines had significantly higher antibodies to these viruses than unvaccinated controls; however, serological responses to killed BVDV vaccines were low. Calves with preexisting antibodies to IBRV, PI3V, BRSV, and the Singer strain of BVDV had lower seroconversion rates following vaccination than calves that were seronegative initially.Serological responses in calves to IBRV, PI3V, BRSV, and BVDV differed among various commercial vaccines. Antibody titers to IBRV were higher in calves vaccinated with modified-live IBRV vaccines than in those vaccinated with killed IBRV vaccines. Following double vaccination with modified live IBRV and PI3V vaccines, seroconversion rates and antibody titers to IBRV and PI3V were higher in calves vaccinated intramuscularly than in those vaccinated intranasally. Calves given Cattlemaster 4 had significantly higher titers to BRSV and PI3V, and lower titers to BVDV, than calves given Cattlemaster 3, suggesting that the addition of BRSV to Cattlemaster 4 caused some interaction among antigens. PMID- 17423914 TI - The seroprevalence of Toxoplasma gondii in Ontario sheep flocks. AB - In a random sample of 103 sheep farms in Ontario, 99% of the farms had some sheep serologically positive for Toxoplasma gondii, based on an enzymelinked immunosorbent assay (ELISA). The percent of sheep affected within farms ranged from 3.8% to 97.8%, with an average flock prevalence of 57.6%. When farm management variables were considered in a multivariate analysis, significantly lower rates of serologically positive sheep were associated with neutering of female cats and clipping of ewes' perineums before lambing; significantly higher prevalence rates were found on farms where sheep were purchased from other flocks, pigs were raised on the same farm, sheep shared pasture with other animals, flowing water was available at pasture, and pastured replacements had access to housing. As well, in univariate analyses, higher prevalence was positively associated with an increasing number of cat litters born over the previous two years and offering creep feed or forage to lambs, and inversely with the amount of labor expended on sheep rearing. PMID- 17423915 TI - Persistent shedding of Salmonella enteritidis from the udder of a cow. AB - Salmonella enteritidis phagetype 8 was isolated from ill humans, milk-line filters, milk from a bulk tank, and milk from the right hind quarter of a five year-old Holstein cow on a dairy farm in southern Alberta. The affected animal was removed from the herd and continued to shed S. enteritidis from this quarter during a seven-month interval in an isolation facility. Milk from the affected quarter was visually normal, and no other pathogen was isolated from the udder during the investigation. After removal of the infected cow from the herd, milk from the bulk tank was culturally negative for Salmonella sp. during the succeeding 15 months. PMID- 17423916 TI - Observations on the influence of exogenous gonadotrophins and cloprostenol on ovulation in gilts. AB - We studied the effects of gonadotrophins and prostaglandin (PG) F(2alpha) on ovulation in gilts. Twenty-eight gilts were induced to ovulate using 750 IU pregnant mares serum gonadotrophin (PMSG) and 500 IU human chorionic gonadotrophin (hCG), administered 72 h apart. At 34 and 36 h after hCG, gilts received injections of either 500 mug or 175 mug PGF(2alpha) (cloprostenol), or had no injections. Laparotomies were performed at 36 h (cloprostenol gilts) or 38 h (controls) after hCG injection. The ovaries were examined and the proportion of preovulatory follicles that had ovulated (ovulation percent) was determined at 30 min intervals for up to 6 h. The number of gilts in which ovulation was initiated and the ovulation percent increased (p<0.001) with time, but was not affected by treatment. Many medium sized follicles ( 0.249) at processing on day 57, was higher in the groups receiving poultry by product and poultry fat: 27(22.5%) in group 2, 26(21.7%) in group 3, and 20(16.7%) in group 4 compared to that of the controls 12(10%). PMID- 17424019 TI - Effects of scrotal circumference of sire on age of calving in Ontario beef heifers. AB - Calving records for daughters of 667 Record of Performance tested sires were examined to study factors related to age at first calving. For the 3025 heifers analyzed, mean age (+/- SD) at first calving was 791.5 +/- 161.6 days (26.4 +/- 5.4 months). There were significant effects of the breed of sire on age of heifer at first calving. Daughters of Angus sires calved earlier than daughters of Charolais, Hereford, Limousin, and Simmental sires. Mean ages at first calving for daughters of Hereford, Limousin, and Simmental bulls were significantly (p < 0.05) correlated with the sire's end-of-test age-adjusted scrotal circumference, but these correlations were not significant when farm effects were controlled. It appears that unknown herd level factors are more important in determining age at first calving in Ontario herds than using early maturing sires as determined by scrotal circumference measurements. PMID- 17424020 TI - Common ocular problems of Shin Tzu dogs. AB - In recent years the Shih Tzu has become a popular breed of dog. It has a number of serious diseases of the eye which are not being recognized by breeders, owners, and sometimes veterinarians. The purpose of this communication is to assist veterinarians in recognizing and treating, where possible, some of the common ocular problems of this breed, including medial canthal entropion, various trichiasis problems, dermoids, third eyelid gland prolapse, chronic keratitis and corneal ulcers, progressive retinal atrophy, vitreal syneresis, and retinal detachment. Because most of the problems are inherited or of suspected genetic origin, breeders should receive genetic counselling from their veterinarian. PMID- 17424021 TI - Effect of antiparasitic treatment in beef replacement heifers. AB - Two studies were conducted in western Canada to evaluate the impact of antiparasitic treatment on weight gain and reproductive performance in beef replacement heifers from weaning to first breeding. Treatment of heifers at weaning with ivermectin increased the rate of weight gain in comparison to treatment with fenthion. The feed efficiency of ivermectin-treated animals was also significantly better than those treated with fenthion. Increased weight was associated with an increased number of animals in estrus at the end of the feeding period for heifers receiving ivermectin. We concluded that treatment with ivermectin improved feed efficiency and weight gain in heifers from weaning to breeding. In turn, this weight gain appeared to be associated with an increased number of heifers in estrus. PMID- 17424022 TI - A field trial to determine the efficacy of long-acting oxytetracycline for reducing the treatment rate in aborted feedlot heifers. PMID- 17424023 TI - An unusual complication of strangles in a pony. PMID- 17424024 TI - Heterotopic bone in the abdominal wall due to metastatic transitional cell carcinoma in a dog. PMID- 17424025 TI - Surgical repair of coxofemoral luxation in a Miniature horse. PMID- 17424026 TI - Quebec. Distribution of Streptococcus suis capsular types in Canada in 1991. PMID- 17424028 TI - British Columbia. Septicemia suspected to be caused by a ricksettia-like agent in farmed Atlantic salmon. PMID- 17424027 TI - Atlantic Canada. Hitra disease in Atlantic salmon. PMID- 17424029 TI - Western Canada. Pseudopositive ELISA results for infectious bovine rhinotracheitis IgG reactors in three artificial insemination studs. PMID- 17424030 TI - Further diagnosis and treatment of canine dominance aggression. PMID- 17424032 TI - Here's tae us, wha's like us? PMID- 17424031 TI - Diagnostic ophthalmology. PMID- 17424033 TI - The toxic potential of antifreeze for animals. PMID- 17424035 TI - CVMA Annual Review. PMID- 17424034 TI - Veterinary medical ethics. PMID- 17424036 TI - CVMA insurance survey questionnaire. PMID- 17424037 TI - Epidemiological associations between characteristics of registered broiler chicken flocks in Canada and the Salmonella culture status of floor litter and drinking water. AB - A Canada-wide flock management survey of 294 randomly selected commercial broiler chicken flocks was conducted during 1989-1990. The prevalence of flocks that yielded Salmonella from cultures of floor litter or drinking water, and the prevalence of floor litter samples that yielded Salmonella, were significantly associated (p < 0.05) with the age of the flock and the region of Canda in which the flock was located. Culture of Salmonella from the drinking water was significantly associated (p < 0.05) with the type of drinker used, the dead bird disposal method, and the region of Canada in which the flock was located. There was a significantly greater risk (p < 0.05) of contamination of drinking water with Salmonella from trough drinkers (odds ratio = 7.99) and plastic bell drinkers (odds ratio = 6.10) than from nipple drinkers. No significant associations were found between pest control, restrictions on visitors, clean-out methods, or water sanitization and the culture of salmonellae from floor litter or drinking water. PMID- 17424038 TI - Abomasal disease in young beef calves: Surgical findings and management factors. AB - A retrospective study was performed on 46 abomasotomies in young beef calves. Clinical abomasal disease in the herds under investigation had an average incidence of 1.0% for the year of the study. The range in incidence among herds was 0.2-5.7%. In 80% (37/46) of abomasal surgeries, abomasal ulcers were found. In 76% (35/46) of the cases, hair was present in the abomasum. These findings indicate that there could be a link between abomasal ulcers and the ingestion of hair by young beef calves.Housing appeared to be a significant factor in relation to the relative risk of abomasal disease. Calves housed in pens or on stubble fields were nearly three times as likely to receive surgery for abomasal disease than were those housed on pasture.The results also indicated that mineral supplements may be helpful in reducing the incidence of abomasal disease. PMID- 17424039 TI - Comparison of the efficacy of three premedicants administered to cats. AB - Healthy cats (n = 90), anesthetized for minor procedures, were included in a study designed to evaluate the efficacy of three premedicant mixtures. The drug combination was assigned randomly and the evaluations were made by individuals unaware of the treatment used. The mixtures and their final concentrations were as follows: acepromazine (1.0 mg/mL) and atropine (0.25 mg/mL) with either meperidine (20.0 mg/mL), ketamine (25.0 mg/mL), or oxymorphone (0.2 mg/mL). The dose used was 0.2 mL/kg(0.75). There was no significant difference (p< 0.05) among drug combinations in the degree of sedation achieved, difficulty of handling for IV catheter placement, induction dose of thiopental, or heart or respiratory rate following induction. All combinations were considered satisfactory for premedication of healthy cats. The ketamine combination had a tendency for more consistent sedation (0.05 < p < 0.01). PMID- 17424040 TI - Calcinosis circumscripta involving the metatarsal region in a dog with chronic renal failure. PMID- 17424041 TI - Serum and urinary levels of salbutamol after chronic oral administration in a calf. PMID- 17424043 TI - Western Canada. Rabies diagnoses in western Canada July 1-December 31, 1991. PMID- 17424042 TI - Surgical correction of uterine inversion in a mare. PMID- 17424044 TI - Ontario. Submandibular and disseminated zygomycosis in pigs. PMID- 17424046 TI - Quelle est la valeur de mon hopital veterinaire? PMID- 17424045 TI - Alberta. Sarcoptic mange in ranch foxes. PMID- 17424047 TI - CVMA should not have GATT involved! PMID- 17424048 TI - Deprofessionalization of the federal public service. PMID- 17424049 TI - New information on "Blue Ear' disease. PMID- 17424050 TI - The value of antimicrobial susceptibility testing. PMID- 17424051 TI - Small animal versus large animal articles in the Canadian Veterinary Journal. PMID- 17424052 TI - Veterinary medical ethics. PMID- 17424053 TI - CVMA Awards 1992. PMID- 17424054 TI - We had a whale of a time! PMID- 17424055 TI - The veterinarian and the human animal bond. PMID- 17424056 TI - Pets in public housing - on breaking the bond. PMID- 17424058 TI - The correlations between cruelty to animals and child abuse and the implications for veterinary medicine. PMID- 17424057 TI - Canine corrections - the human animal bond behind bars. PMID- 17424059 TI - The mystery at the top of the leash: Reading clients in bad times. PMID- 17424060 TI - Antibiotic residues (bacterial inhibitory substances) in the milk of cows treated under label and extra-label conditions. AB - The objectives of this study were to describe the depletion pattern of antibiotic residues (microbial inhibitory substances) from the milk of cows treated under field conditions of clinical disease and antibiotic administration, including both label and extra-label use, and to determine if the type of extra-label use, the route of administration, and the drug used were factors associated with prolonged shedding of residues in milk.Milk samples from 138 cows, treated with a variety of antibiotic products on farms in southwestern Ontario in 1989 and 1990, were collected before treatment and for six days after cessation of treatment. Samples were tested for antibiotic residues with the Brilliant Black reduction test (BR-test) and the Bacillus stearothermophilus var calidolactis disc assay. In 13/138 (9.4%) of cow treatments, at least one milk sample was positive on both antibiotic residue tests after the label milk withholding time for the drug(s) used. In ten of these instances, the antibiotics were administered in extra-label fashion and in three the drugs were reportedly used according to label instructions. Extra-label use of antibiotics was significantly associated with increased risk of antibiotic residues in milk beyond the label withholding time. No significant differences in risk were observed among the various antibiotic products used in the study.The farms involved in this study were selected on the basis of their proximity to our laboratory; therefore, the frequency of antibiotic residue detection after withholding times may not be indicative of the provincial or national situation. If farmers and veterinarians find themselves in a situation where extra-label use of an antibiotic is necessary, use of an alternative antibiotic that can be used at label dose and with a known withdrawal time may avoid a problem with residues. PMID- 17424061 TI - Roquefortine in the stomach contents of dogs suspected of strychnine poisoning in Alberta. AB - From April to September 1990, submissions in Alberta veterinary diagnostic laboratories for which strychnine analysis was requested were tested retrospectively for roquefortine, a diketopiperazine alkaloidal tremorgenic mycotoxin. Roquefortine was found only in strychnine-negative samples. The specific origins of the fungal-contaminated specimens could not be determined. Of the six roquefortine-positive cases, four dogs that vomited prior to treatment recovered. Two dogs which died had significant amounts of stomach contents present at the time of postmortem.At the present time only one veterinary laboratory in Canada is routinely testing for roquefortine when samples are negative for strychnine. It appears that a low diagnostic rate for this type of poisoning may be occurring due to incomplete testing. The inability to differentiate between roquefortine and strychnine poisoning on a clinical basis in five of our six cases emphasizes that an accurate causative diagnosis requires laboratory examination.Research in rats and sheep has shown that the tremorgenic mycotoxins penitrem A and roquefortine are excreted through bile. Although further research is required, the submission of bile and intestinal contents is recommended if stomach contents or vomitus are not available for laboratory testing. Both of these mycotoxins should be tested for when strychnine analysis is negative as fungi may produce both toxins at the same time. In this study we were unsure if roquefortine alone or in combination with other toxins was responsible for our findings. PMID- 17424062 TI - Fatal herpesvirus infection in commercial rabbits. AB - Acute mortality occurred in two unrelated rabbitries. In the rabbits examined, an unidentified herpesvirus caused lesions that have not been reported previously in this species. The primary lesions were multifocal hemorrhagic dermatitis on the face and back, localized pneumonia, and severe splenic necrosis. Large eosinophilic, intranuclear inclusion bodies that were observed in tissue sections of skin, spleen, and lung were identified as herpes-like viral particles by electron microscopy, and herpesvirus was cultured on rabbit kidney cells. Intramuscular injection of tissue culture fluid containing virus resulted in mortality and severe illness in two seven-week-old domestic rabbits four and six days postinfection, respectively. The gross and microscopic lesions were reproduced and herpeslike viral inclusions were observed in skin lesions. Herpesvirus was recovered from lung, trachea, spleen, liver, and from the thigh muscle at the site of inoculation. The experimental infection also activated severe pasteurella septicemia. The herpesvirus isolate needs further characterization. PMID- 17424063 TI - Agenesis of the cecum and the ascending and transverse colon in a 12-yer-old cat. PMID- 17424064 TI - Mycotic dermatitis in ostriches. PMID- 17424065 TI - Concentrations of copper in tissues of wapiti raised in Saskatchewan. PMID- 17424066 TI - Prince Edward Island. Interstitial pneumonia in piglets. PMID- 17424067 TI - Quebec. Isolation of the porcine reproductive and respiratory syndrome virus in Quebec. PMID- 17424069 TI - British Columbia. Lead poisoning in Trumpeter swans. PMID- 17424068 TI - Quebec. Isolation of Cryptococcus neoformans from bovine milk. PMID- 17424070 TI - Diagnostic ophthalmology. PMID- 17424071 TI - 2000 A.D. and all that. PMID- 17424072 TI - Council takes important steps in St. John's. PMID- 17424073 TI - Career options for veterinary students graduating in 2000 A.D. - A western Canadian viewpoint. PMID- 17424074 TI - The use of magnetic resonance imaging in the diagnosis of neurological disease. AB - Magnetic resonance imaging (MRI) data were correlated with clinical and cerebrospinal fluid (CSF) findings in one cat and two dogs with brain lesions. In all three cases, localization of the lesions, as determined clinically, was confirmed using MRI. Magnetic resonance imaging also helped us to define the full extent of the lesion(s) in each case. In one case, the lesion would have been diagnosed as purely inflammatory based on the abnormalities in the CSF. The MRI study, however, showed a homogeneous mass with circumferential changes characteristic of peritumoral edema or inflammation. In two cases, the MRI findings were confirmed at necropsy. An MRI study was also done on a normal dog, demonstrating the variable contrast and anatomical detail possible using this technique. We also discuss difficulties in identifying tumor type using MRI. PMID- 17424075 TI - Chronic obstructive pulmonary disease: Usefulness of clinical signs, bronchoalveolar lavage, and lung biopsy as diagnostic and prognostic aids. AB - We examined 18 horses with clinical signs of chronic obstructive pulmonary disease (COPD) using physical examination, cytological and bacteriological evaluation of bronchoalveolar fluid, and percutaneous lung biopsy. In 16 cases, histological examination of lung tissue confirmed the diagnosis of COPD. Two horses were excluded: one had uncomplicated bacterial pneumonia and in the other a satisfactory lung biopsy could not be obtained. In horses with COPD, the most common historical complaint was coughing, which was reported in 88%. The most frequently detected abnormal finding on physical examination was abnormal lung sounds; these were detected in 69% of horses at rest and in 88% of horses breathing deeply into a bag. A novel finding was that 29% of horses had lung sounds that were quieter than would be expected for the degree of respiratory effort. Horses with COPD had increased percentages of neutrophils and decreased percentages of lymphocytes and macrophages in their bronchoalveolar lavage fluid. Bronchiolar neutrophil infiltration and peribronchiolar mast cell accumulation in lung biopsy tissue had the highest correlation with clinical condition. The severity of pathological changes in biopsies of lung did not predict whether the horse would die in the two to four year follow-up period. Horses that died in the follow-up period were more severely affected clinically at initial presentation than horses that were alive at the end of the follow-up period. PMID- 17424076 TI - Comparison of a combination of oxfendazole and fenthion versus ivermectin in feedlot calves. AB - A trial involving 6,169 feedlot calves was conducted under commercial feedlot conditions in western Canada to compare the relative efficacy of treatment with a combination of oxfendazole and fenthion (O/F) versus ivermectin (I) with respect to the outcome variables, final weight, gain, days on feed (DOF), dry matter intake (DMI), average daily gain (ADG), dry matter intake to gain ratio (DM:G), and morbidity, mortality, and carcass grade parameters.There were no significant differences (p >/= 0.05) between the treatment groups for final weight, gain, DOF, DMI, ADG and DM:G. In addition, there were no significant differences (p >/= 0.05) in the carcass grading parameters between the treatment groups. The bovine respiratory disease (BRD) relapse rates, the overall mortality rates, and the cause specific mortality rates were not significantly different (p >/= 0.05) between the treatment groups. The BRD treatment rate in the O/F group was significantly lower (p /= 1:8 were considered positive. The overall prevalence in 267 sows in various groups was 37.8% (range 20-86%). The prevalence in sows maintained in total confinement was significantly lower than in sows not maintained in total confinement (p = 0.01). Prevalence in sows was not associated with other herd characteristics or with abundance estimates of rats and/or mice.Free-living animals tested included 74 domestic cats (Felis domestica), 203 house mice (Mus musculus), 15 mice of the genus Peromyscus, nine Norway rats (Rattus norvegicus), 34 opossums (Didelphis virginiana), 14 raccoons (Procyon lotor), and seven striped skunks (Mephitis mephitis). Of these, the only seropositive animals were two domestic cats.The results of this study failed to implicate the free-living species surveyed as important reservoirs of EMCV for swine, and suggested that swine themselves and/or some species not included in this survey are the main reservoir of EMCV for swine in Iowa. PMID- 17424089 TI - Low energy laser irradiation treatment for second intention wound healing in horses. AB - Low energy helium-neon laser irradiation was administered to full thickness skin wounds (3 cm x 3 cm) on the dorsal surface of the metacarpophalangeal/metatarsophalangeal joints and cranial surface of the tarsocrural joints of eight horses. The effects on wound healing were analyzed statistically. There were no differences (p > 0.55) observed in the rate of wound healing between the low energy laser irradiated wounds and the control wounds. There was a significant difference (p < 0.006) observed in the rate of healing between the anatomical sites. Tarsal wounds healed more rapidly than fetlock wounds. PMID- 17424090 TI - Urinary tract ultrasonography in normal rams and in rams with obstructive urolithiasis. AB - We determined the position, dimensions, and structure of the kidneys, ureters, bladder, and urethra in 20 healthy, adult rams by use of ultrasonography. The findings were compared with those of seven rams with obstructive urolithiasis, thus establishing criteria for the diagnosis of urolithiasis via ultrasonography. A 5.0 MHz convex transducer was placed over the right paralumbar fossa to examine the kidneys, and a 5.0 MHz linear rectal transducer was used to examine the bladder and urethra transrectally. All examinations were performed on standing rams. The left kidney had a length of 8.4 +/- 0.3 cm (mean +/- SD), a width of 4.7 +/- 0.3 cm, and a depth of 4.4 +/- 0.3 cm. The diameter of the renal sinus of the left kidney was 1.5 +/- 0.2 cm. The circumference of the medullary pyramids measured 2.8 +/- 0.3 cm. Similar ultrasonographic measurements were obtained for the right kidney. The mean diameter of the bladder of 12 rams was 7.5 +/- 2.8 cm. The diameter of the bladder could not be determined in the remaining eight rams because it was greater than 10 cm and therefore beyond the penetration depth of the scanner. The only part of the urethra which could be visualized ultrasonographically was the internal urethral orifice. It had a diameter of 0.2 +/- 0.1 cm.Ultrasonographic examination of seven rams with obstructive urolithiasis revealed a markedly dilated urethra and urinary bladder. Due to severe cystitis, the contents of the urinary bladder appeared as multiple, tiny, uniformly distributed echoes. The renal pelvis and medullary pyramids of both kidneys were dilated in four rams. In two rams, uroperitoneum and accumulation of urine in the abdomen were diagnosed via ultrasonography. In one ram this was due to a ruptured ureter and in one to a ruptured bladder. The results of this study indicate that ultrasonography is a useful aid in the diagnosis of obstructive urolithiasis. PMID- 17424091 TI - Intussusception in dogs and cats: A review of 36 cases. AB - Intussusception is recognized as a common cause of bowel obstruction in small animals. This study documents the clinical and surgical findings in nine cats and 27 dogs diagnosed as having intussusception. The main purposes of the study were to define the predisposing causes and clinical signs of intussusception and to evaluate various surgical techniques commonly employed in its treatment. No common predisposing cause could be established. Diagnosis of intussusception was based most often on clinical signs of bowel obstruction in association with the palpable abdominal mass. The majority of the intussusceptions involved the enterocolic junction. Formation of adhesions was more frequent in cats. Surgical treatments included simple reduction, manual reduction with plication, intestinal resection/anastomosis, and intestinal resection/anastamosis with plication. There was no statistically significant difference (p>0.05) in the recurrence rate of the intussusceptions when the various surgical techniques were compared. Recurrence of an intussusception was not related to either the bowel segment involved or whether a simple reduction, bowel resection, or intestinal plication was performed at the initial surgery. PMID- 17424092 TI - The effect of levels of concentrate in feedlot diets on the health status of beef calves. AB - In this two year study we investigated the effect that the level of concentrate in feedlot diet had on the occurrence of health problems in bull calves belonging to two multibreed groups.In each of the two years, the 168 day feedlot period was divided into two equal periods of 77 days with an intervening 14 day adjustment period. During the first period, the animals in half of the pens within each breed-group received a high concentrate (85% grain) diet (H) while the rest of the pens received a diet (L) of hay ad libitum and half as much concentrate as the bulls on high concentrate. The L diet contained an average of 48% grain. In the second period, diets of bulls in half of the pens within each breed-group were switched from L to H and vice versa. During the feedlot period, the incidence of sickness among bulls was recorded. Disease incidence was higher in the second year compared to the first. In the second year, 49 out of 56 (87.5%) calves on the high concentrate diet required treatment for respiratory infections compared to only four (7.14%) in bulls on the low concentrate diet. The results suggest that a high concentrate diet for animals in feedlots may be more stressful to calves coming directly to the feedlot following weaning compared to a low concentrate diet, thereby acting as an additional predisposing factor to respiratory infections. PMID- 17424093 TI - Lymphosarcoma causing acquired obstructive hydrocephalus in a dog. PMID- 17424094 TI - Polycythemia vera and glomerulonephritis in a dog. PMID- 17424095 TI - Detection of right atrial hemangiosarcoma using nonselective angiocardiography in a dog. PMID- 17424096 TI - Ontario. Braxy-like abomasitis in a calf. PMID- 17424098 TI - Manitoba. Salinomycin toxicosis in turkeys: Serum chemistry as an aid to early diagnosis. PMID- 17424097 TI - Prince Edward Island. Salmonella heidelberg in a dairy herd. PMID- 17424099 TI - British Columbia. Fenthion toxicity in bald eagles. PMID- 17424100 TI - Managing your fee schedule. PMID- 17424103 TI - Support for GATT involvement. PMID- 17424102 TI - Diagnostic ophthalmology. PMID- 17424104 TI - CVMA GATT involved on behalf of members. PMID- 17424105 TI - Quantitative analyses of field data in private practice: A caveat. PMID- 17424106 TI - Deprofessionalization within Agriculture Canada: A manager's perspective. PMID- 17424107 TI - Simplest answer is the most probable answer. PMID- 17424108 TI - Public relations that money can't buy. PMID- 17424109 TI - Meaning versus mechanism. PMID- 17424110 TI - Elitism damages the professional image. PMID- 17424111 TI - Veterinary medical ethics. PMID- 17424112 TI - Career options for veterinary graduates in the year 2000: An Ontario perspective. PMID- 17424113 TI - Haemophilus somnus infection: A retrospective analysis of cattle necropsied at the Western College of Veterinary Medicine from 1970 to 1990. AB - Results of necropsy examinations of 297 bovine carcasses with naturally occurring disease caused by Haemophilus somnus infection, and necropsied at the Western College of Veterinary Medicine in the period from 1970 to 1990, were analyzed with reference to the ages of affected cattle, seasonal occurrence of disease, the relative frequency of encephalitis, pneumonia, and myocarditis, and interrelationships among these conditions. The cattle came from 177 separate herds in the province of Saskatchewan. There was a range in age from one week to 10 years, and a mean age of 38 weeks. The diseases had a marked seasonal fall and winter incidence. The data indicated a trend characterized by an increasing percentage of cattle with pneumonia and myocarditis and a decreasing percentage with encephalitis during the period. PMID- 17424114 TI - Prevalence of feline immunodeficiency virus in submissions of feline serum to a diagnostic laboratory in Atlantic Canada. AB - The purpose of this project was to identify the prevalence of feline immunodeficiency virus (FIV) in the Atlantic region of Canada, and to determine possible associations between FIV serological status and breed, sex, and age. Feline serum samples (671) submitted to the Prince Edward Island Diagnostic Services - Atlantic Veterinary College laboratory between January 1, 1988 and July 30, 1989 were considered eligible for this study. The majority of samples originated from Prince Edward Island (607). Testing was performed in duplicate using commercial 96-well enzyme-linked immunosorbent assay test kits for FIV antibody. Results included a seropositive rate of 7.6% for all submissions. Mean age of FIV-seropositive cats was eight years. There was an increasing risk of FIV seropositive status associated with age. Prevalence of FIV among intact males was significantly higher (odds ratio = 2.59) than other gender categories. The principal conclusion of this study was that FIV is present in cats of the Atlantic provinces, and that its associations and prevalence are consistent with those found in other North American epidemiological studies. PMID- 17424115 TI - Porcine respiratory coronavirus in Quebec: Serological studies using a competitive inhibition enzyme-linked immunosorbent assay. AB - Porcine respiratory coronavirus (PRCV) was identified for the first time in Quebec, using a blocking enzyme-linked immunosorbent assay (ELISA). Unlike the virus neutralization test (VNT), this ELISA was able to distinguish transmissible gastroenteritis virus (TGEV) from PRCV. Among the 15 seropositive fattening herds from group A, sera containing PRCV antibodies represented 74.8%, whereas those with TGEV antibodies represented only 7.2%. In group B, which consisted of 15 sow herds, nine herds expressed only PRCV-specific antibodies while the other herds had animals positive for TGEV-specific antibodies. PMID- 17424116 TI - The effect of subunit or modified live bovine herpesvirus-1 vaccines on the efficacy of a recombinant Pasteurella haemolytica vaccine for the prevention of respiratory disease in feedlot calves. AB - The efficacy of a Pasteurella haemolytica vaccine (PhV) administered once to calves within 24 hours of arrival at a feedlot was tested for the ability to prevent morbidity and mortality from all bovine respiratory disease (BRD) and specifically from fibrinous pneumonia mortality. The PhV consisted of two immunizing ingredients: outer membrane proteins extracted from P. haemolytica, plus genetically attenuated leukotoxin produced by recombinant DNA technology. This double blind study was conducted at a large Saskatchewan feedlot using 2,324 high-risk calves purchased at auction markets and kept under typical commercial feedlot conditions. The trial design included four vaccine test groups: 1) PhV and a bovine herpesvirus type-1 (BHV-1) subunit vaccine comprised only of the virus glycoprotein IV (gIV); 2) PhV and a commercial modified live vaccine (MLV) containing BHV-1 and parainfluenza-3 viruses; 3) gIV alone; and 4) MLV alone. Calves were assigned to vaccine groups in a random systematic manner, individually identified, and monitored for 90 days after vaccination. The vaccines were given once, on arrival, to reflect common feedlot practice, although vaccination prior to expected risk would be more appropriate.The PhV in combination with gIV reduced BRD morbidity by 20% (p < 0.05) compared to gIV alone and 24% (p < 0.05) compared to MLV alone, and reduced BRD mortality by 88% (p < 0.05) and fibrinous pneumonia mortality by 100% (p < 0.05) when compared to either gIV or MLV alone. Vaccination with PhV in combination with MLV significantly reduced the efficacy of the PhV in preventing BRD morbidity, BRD mortality, and fibrinous pneumonia mortality and also reduced the antibody response to P. haemolytica leukotoxin. These results suggest that the MLV interfered with the protective capacity of the PhV. PMID- 17424117 TI - A survey of house mice from Iowa swine farms for infection with Leptospira interrogans serovar bratislava. PMID- 17424118 TI - Isolation of Gardnerella vaginalis from the genital tract of six mares. PMID- 17424119 TI - Lameness in breeding age swine - A case study. PMID- 17424120 TI - Monensin/roxarsone contaminated dog food associated with myodegeneration and renal medullary necrosis in dogs. PMID- 17424121 TI - Surgical correction of cystic urolithiasis in an iguana. PMID- 17424122 TI - Cytodites nudus-induced granulomatous pneumonia in chickens. PMID- 17424123 TI - Ontario. Menadione (vitamin K(3)) toxicity in six horses. PMID- 17424124 TI - Labrador. Rabies epizootic in Labrador. PMID- 17424125 TI - Western Canada. Western Canada rabies diagnoses January 1-June 30, 1992. PMID- 17424126 TI - Diagnostic ophthalmology. PMID- 17424128 TI - Injection site residues of benzathine/procaine penicillin G. PMID- 17424130 TI - Career options for veterinary students graduating in the year 2000 A.D.: An Atlantic provinces perspective. PMID- 17424129 TI - Veterinary medical ethics. PMID- 17424131 TI - Meta-analysis of field trials of antimicrobial mass medication for prophylaxis of bovine respiratory disease in feedlot cattle. AB - One hundred and seven field trials of prophylactic mass medication for bovine respiratory disease (BRD) in feedlot cattle were reviewed. Meta-analysis is the formal quantitative statistical review process that was used to synthesize the data from randomized field trials and draw conclusions concerning the efficacy of prophylactic mass medication in feedlot calves.The results of the meta-analysis indicated that prophylactic parenteral mass medication of calves with long-acting oxytetracycline or tilmicosin on arrival at the feedlot would reduce BRD morbidity rates (p < 0.001). There were, however, unreliable data on the effects of mass medication on mortality rates and performance, insufficient data on the most effective treatment regimes, and no valid data on the efficacy of feed and water medication for prophylaxis of BRD.This review highlights the gaps in our knowledge and points out the need for additional well-designed randomized controlled field trials of adequate size to assess the efficacy and socioeconomic impact of prophylactic mass medication for BRD in feedlot cattle. PMID- 17424132 TI - Paramyxovirus-1 in feral pigeons (Columba livia) in Ontario. AB - Paramyxovirus-1 (PMV-1) infection was diagnosed in racing pigeons in Ontario during 1985, but it was not until January 1989, that the virus was isolated from feral pigeons (Columba livia) in this province. During an 18 month period beginning January 1988, a total of 43 feral pigeons was submitted to the Wildlife Diseases Laboratory, Pathology Department, Ontario Veterinary College. A history of neurological signs accompanied most of the birds. Tissues from 29 birds were submitted for PMV-1 isolation. Allantoic inoculation of embryonated chicken eggs yielded PMV-1 in 10 of the pigeons submitted. On the basis of histological criteria, we believe that 12 other birds were also infected with PMV-1.Gross pathological changes were unremarkable. Lymphplasmacytic interstitial nephritis was observed histologically in all birds from which PMV-1 was isolated. Other lesions seen, in decreasing frequency of occurrence, were lymphoplasmacytic interstitial hepatitis and multifocal hepatic necrosis, lymphoplasmacytic interstitial pancreatitis, nonsuppurative encephalitis and myelitis.The existence of PMV-1 in feral pigeons poses a potential threat to the poultry population since there is ample opportunity for mingling with poultry under open housing management. There is also a concern that pigeons may harbor the virus, perhaps in the kidney, and become chronic carriers and potential long-term disseminators of the disease. PMID- 17424133 TI - Swine reproductive and respiratory syndrome in Quebec: Isolation of an enveloped virus serologically-related to Lelystad virus. AB - Sera were collected from convalescent sows and sick piglets from six pig farms in southern Quebec that have experienced outbreaks of the so-called porcine reproductive and respiratory syndrome. By indirect immunoperoxidase, a few of these sera (4 of 14) (28.6%) were found to be positive for antibody to the Lelystad virus, whereas by indirect immunofluorescence 30 of 36 (83.3%) were positive for antibody to the antigenically-related American isolate ATCC-VR2332. Pregnant sows inoculated intranasally with filtered homogenates prepared from the lungs of necropsied piglets obtained from a seropositive farm developed fever, inappetence, and reproductive failure characterized by stillbirths and various stages of mummification. Lesions of interstitial pneumonia were induced in experimentally-infected specific pathogen-free piglets. A virus, having morphological and biological characteristics of viruses assigned to the family Togaviridae, was isolated from lung tissues of experimentally-infected animals; it could only be propagated in primary cultures of porcine alveolar macrophages. Identification of the virus was confirmed by indirect immunofluorescence using a monoclonal antibody directed against the nucleocapsid protein of the ATCC-VR2332 isolate and porcine sera that were found positive for antibody to both the Lelystad and ATCC-VR2332 isolates. PMID- 17424134 TI - Dysuria and stranguria associated with colonic ulceration and abdominal abscess in a horse. PMID- 17424135 TI - Bovine coronavirus infection in Ontario 1990-1991. PMID- 17424136 TI - Lead toxicosis and changes in the blood lead concentration of dogs exposed to dust containing high levels of lead. PMID- 17424138 TI - Ontario. Persistent sporotrichosis in a dog. PMID- 17424137 TI - British Columbia. Mastitis caused by a Mycobacterium sp. PMID- 17424139 TI - Saskatchewan. Tyzzer's disease in a dog. PMID- 17424140 TI - Veterinary medical ethics. PMID- 17424141 TI - A clinical trial on the efficacy of clemastine in the management of allergic pruritus in dogs. AB - Clemastine fumarate was administered to 72 atopic dogs. The pruritus in seven dogs was eliminated with this treatment. Another 14 dogs improved considerably with treatment but had some residual pruritus. Side effects were uncommon and only three dogs had to be withdrawn from treatment because of adverse reactions. PMID- 17424142 TI - CRITTER: A database for managing research animals. AB - We describe CRITTER, a computer database program for managing research animals. We designed it especially for institutions which operate health surveillance plans, such as specific pathogen-free schemes. Because CRITTER can be used to record any type of test result in any species of animal, it can be customized to suit each institution and its management protocol. In addition to maintaining a current inventory of each individual animal and its location, the program retains historical information on those that have been removed from the colony. Output summaries are generated by selecting from a menu of standard reports or by designing a custom query. Although CRITTER has been designed for individual research establishments, it could be modified for use in area health surveillance programs. CRITTER operates on IBM compatible computers using a menu-driven, runtime version of Paradox. PMID- 17424143 TI - Monitoring pregnancy losses in small dairy herds. PMID- 17424144 TI - Acute myelomonocytic leukemia in a horse. PMID- 17424145 TI - Disposition of feedlot heifer and beef carcasses under a Canadian streamlined inspection system. PMID- 17424146 TI - Acute alcohol intoxication in a dog. PMID- 17424148 TI - The veterinary profession - A service industry. PMID- 17424147 TI - A study of pituitary and adrenal function in a disease-stressed ram. PMID- 17424149 TI - Questions about Hitra disease in Atlantic salmon. PMID- 17424150 TI - Personnel plumbing. PMID- 17424151 TI - Employment contracts and restrictive covenants. PMID- 17424152 TI - Ontario. Death due to Clostridium perfringens type D enterotoxemia in adult ewes. PMID- 17424153 TI - Alberta. Ascarid larval pneumonia in pigs. PMID- 17424154 TI - British Columbia. Management-related piglet mortality. PMID- 17424155 TI - Additional information about the occurrence of Hitra disease in Atlantic salmon. PMID- 17424156 TI - Insure to ensure. PMID- 17424158 TI - Wanted: Disability benefits. PMID- 17424157 TI - ELISA reliable for detection of antibodies to BHV1. PMID- 17424159 TI - Recognized and potential variations in the ELISA developed for detecting antibodies to BHV1. PMID- 17424160 TI - Limitations of automobile insurance. PMID- 17424161 TI - No-fault insurance and your veterinary vehicle. PMID- 17424162 TI - A survey of the causes of mortality in adult mink, with emphasis on the lactation period. AB - A study of the pattern and relative frequency of diseases in adult female mink during the lactation period was undertaken. All adult females that died between parturition (April/May) and July 1, 1990, from 48 farms in southern Ontario were selected for study, and the cause of death was determined by gross necropsy. In addition, the cause of death was determined by gross necropsy for all adults and weaned kits that died on one farm between April 1988 and March 1989.The mortality rate among farms in the 1990 study, for adult females during the lactation period, ranged from 0.2% to 10.1%, with a median of 1.9%. Nursing disease (56%) was the most common diagnosis, followed by mastitis (11%), metritis (8%), and dystocia (7%). Escherichia coli and Staphylococcus spp. were the most frequent isolates from the cases of mastitis. In the 1988/1989 study, the mortality rate was highest from May to July, with a large increase in June as a result of nursing disease. PMID- 17424163 TI - Variables related to the progress of parturition and probability of stillbirth in swine. AB - Sow and piglet variables related to probability of stillbirth and to viability score were analyzed in litters from 98 multiparous Yorkshire sows. Immediately after the birth of each piglet, viability was scored using Randall's method. Sow variables related to the probability of stillbirth were average birth weight of the litter (p = 0.0001), sow age (p = 0.001), sow condition score (p = 0.003), length of gestation (p = 0.005), and number of piglets in the litter (p = 0.01). Sow variables related to average viability score were average birth weight of the litter (p = 0.001), standard deviation in birth weight in the litter (p = 0.02), sow age (p = 0.03), sow condition score (p = 0.03), and length of gestation (p = 0.03). Piglet variables related to probability of stillbirth were piglet hemoglobin (p = 0.0001), position in the birth order (p = 0.0001), broken umbilical cord (p = 0.0004), and preceding birth interval (p = 0.0004). Piglet variables related to viability score were piglet hemoglobin (p = 0.0001), position in the birth order (p = 0.0001), broken umbilical cord (p = 0.0001), preceding birth interval (p = 0.0001), and birth weight (p = 0.004). Preceding birth interval was related to whether the piglet was live or stillborn (p = 0.0001), to position in the birth order (p = 0.003), and to the sex of the piglet (p = 0.03).The results demonstrated that sow and piglet variables were highly correlated to probability of stillbirth and to viability score. In addition it was also found that the probability of stillbirth was not associated with the duration of farrowing but with the number of piglets in the litter and piglet hemoglobin level. This study also found that lower weight piglets tend to have poor viability, but are not more prone to stillbirth as commonly suggested. These low viability piglets may survive if the necessary care is given during the farrowing process. PMID- 17424164 TI - Cryptosporidium species in imported ostriches and consideration of possible Implications for birds in Canada. PMID- 17424165 TI - Antemortem diagnosis and attempted treatment of (Halicephalobus) Micronema deletrix infection in a horse. PMID- 17424166 TI - L'hemoglobine glycosylee chez la vache laitiere en periode peripartum. PMID- 17424167 TI - A case of cutaneous sterile pyogranuloma/granuloma syndrome in a golden retriever. PMID- 17424169 TI - Eastern Canada. Eastern Canada rabies diagnoses, January 1 - June 30, 1992. PMID- 17424168 TI - Ontario. Lungworm (Crenosoma vulpis) infection in dogs. PMID- 17424170 TI - Alberta. Strychnine toxicity in livestock. PMID- 17424172 TI - Anteing up what the market will bear. PMID- 17424173 TI - Computer data base for the coccidia. PMID- 17424174 TI - Veterinary medical ethics. PMID- 17424175 TI - New developments regarding transportation of dangerous goods. PMID- 17424176 TI - Edited excerpts from The 1992 Report on Veterinary Specialization in Canada. PMID- 17424177 TI - The enteritis complex in domestic rabbits: A field study. AB - A study of the causative agents of enteritis in domestic rabbits from 44 different accessions is described. In descending order of frequency, the organisms most commonly demonstrated were intestinal and hepatic coccidia (Eimeria species), Escherichia coli, Clostridium spp., Salmonella, Bacillus piliformis, and rotavirus. The species of Eimeria identified included those moderately pathogenic and coccidia of low pathogenicity. Using seven antisera against known enterpathogenic strains of E. coli, only one strain, O15, was identified in three cases. Clostridium perfringens or C. spiroforme was demonstrated in the intestinal contents in 11 cases, and lesions compatible with clostridial enteropathy were identified on gross and histopathology. In a serological survey, over 50% of 200 fryer rabbits submitted to Ontario abattoirs and of animals from commercial rabbitries had detectable antibody to rotavirus, indicating the widespread distribution of rotaviral infections in this species. In the cases of enteritis studied, two or more potentially pathogenic organisms were frequently identified, emphasizing that several different organisms may be acting in concert to produce clinical disease. PMID- 17424178 TI - Shortfalls in commentaries on canine aggression. PMID- 17424180 TI - Objection to CVMA-GATT involvement misunderstood. PMID- 17424179 TI - No place for aggressive behavior by pet dogs. PMID- 17424181 TI - Veterinary medical ethics. PMID- 17424182 TI - Polioencephalomalacia in cattle consuming water with elevated sodium sulfate levels: A herd investigation. AB - Polioencephalomalacia (PEM), hereafter used to refer to the specific lesion of cerebrocortical necrosis, developed in 11 of 110 mature cattle on pasture in central Saskatchewan. The primary water source contained a markedly elevated level of sodium sulfate (7200 ppm). The significant clinical findings of the herd investigation included depression, ataxia, cortical blindness, dysphagia, and death. Diagnosis of PEM was confirmed by histopathological evidence of cerebrocortical and subcortical necrosis with microvascular fibrinoid necrosis predominantly in the thalamic region of three affected cattle. The histopathology of sulfate-associated PEM observed in this herd appears to be unique and its features are presented and discussed. Mean levels for serum transketolase, copper, red blood cell transketolase activity, and thiamine (vitamin B(1)) in all exposed young (n = 100) and mature (n = 99) animals did not reveal evidence of deficiencies. Although the blood thiamine status of the seven surviving, affected animals was not evaluated before treatment with exogenous thiamine, 199 members of the herd had blood thiamine levels within the reference range at the time of the outbreak. The outbreak resolved after cattle were moved to a water source containing acceptable levels of sodium sulfate. PMID- 17424183 TI - Mortality in mink kits from birth to weaning. AB - In 1988, a necropsy survey of the pattern and major causes of mortality in mink kits from birth to weaning was undertaken. The overall preweaning mortality rate was 20%. Mortalities occurring within the first three days after birth accounted for 91% of submissions, and 78% of the kits in this age group had no lesions or bacterial isolates. The average weight of kits which died within one day of birth (7.9 g) was significantly lower than the average birthweight of healthy kits (10.7 g). In kits under four days of age and with lesions, the most common diagnoses were dystocia (12%), systemic infection (4%), anasarca (2%), and congenital defects (1%). In unweaned kits four days of age or older, the most common diagnoses were systemic infection (19%), external trauma (6%), dystocia (5%), and cervical adenitis (2%). PMID- 17424184 TI - Failure of papyerine hydrocholoride and doxycycline hyclate as antipruritis agents in pruritic dogs: Results of an open clinical trial. AB - Papaverine hydrochloride (150 to 300 mg/dog every 12 hours) and doxycycline hyclate (3 mg/kg every 12 hours) were both administered orally as individual agents to 13 dogs with pruritus of allergic or idiopathic origin. No dog improved. Depression, anorexia, and vomiting were seen in one dog treated with papaverine hydrochloride. Under the conditions of the study, papaverine and doxycycline were not useful antipruritic agents for the dog. PMID- 17424185 TI - Partial resection of the large colon for the treatment of stenosis associated with verminous colitis in a horse. PMID- 17424186 TI - Distribution of the different capsular types of Streptococcus suis in nineteen swine nurseries. PMID- 17424187 TI - Tracheal foreign body in a cat. PMID- 17424188 TI - Mycotic meningoencephalitis in a northern pike (Esox lucius). PMID- 17424189 TI - Encephalitis attributed to larval migration of Baylisascaris sp. in emus. PMID- 17424191 TI - Ontario. Newcastle disease in cormorants. PMID- 17424190 TI - Ontario. Impaction in a rhea. PMID- 17424192 TI - Alberta. Infectious laryngotracheitis in commercial broiler chickens. PMID- 17424194 TI - Practitioners' corner. PMID- 17424195 TI - A system of veterinary care of wild animals. PMID- 17424196 TI - Publications related to veterinary medicine by Canadian first authors in 1990. PMID- 17424197 TI - Veterinary medical ethics. PMID- 17424198 TI - Serum bile acids and the assessment of hepatic function in dogs and cats. AB - Current literature in veterinary internal medicine regarding the clinical use of the serum bile acids test to assess hepatobiliary function in dogs and cats is reviewed. The test is best used in cases where clinical signs and routine laboratory tests are suggestive of liver disease. It is a highly sensitive and specific test of hepatic function, and is the best method of assessing liver function available to the private practitioner. Abnormal results do not determine etiology, severity, or prognosis of the disorder. They merely indicate the need for hepatic biopsy. The serum bile acids concentration should always be measured in both a fasting and a two-hour postprandial sample. PMID- 17424199 TI - Congenital spinal stenosis and dam mortality associated with feeding moldy cereal straw. AB - We describe herein an epidemiological investigation of the cause of a syndrome of congenital spinal stenosis (CSS) in four beef herds in western Canada. Three of the herds were affected with the syndrome in 1987, the fourth was affected in 1990. The prevalence of CSS in affected groups of calves varied from 29% (16/55) to 100% (14/14). All affected calves had congenital posterior paralysis or paresis; some calves also had one or more of the following: shortened limbs, forelimb varus deformities, superior brachygnathia, and a dome-shaped cranium. Affected calves showed focal premature closure of axial and appendicular metaphyseal growth plates. At three of the four farms most of the pregnant cows were affected with alopecia with or without pruritus in January, and 25% of the cows in one herd died during the winter. The investigation indicated that CSS was associated with feeding moldy cereal straw to pregnant beef cows during the winter. At all four farms, the cereal straw bales were thoroughly soaked by rain prior to stacking, and obvious mold was present when they were broken open for feeding. Species of both Penicillium and Fusarium were abundant within the bales. The most likely cause of the disease was a fungal mycotoxicosis, although the mycotoxin responsible was not isolated. PMID- 17424200 TI - Surgical repair of rectovaginal fistulae in mares: Twelve cases (1983-1991). AB - The case records of 12 mares at the Ontario and Atlantic Veterinary Colleges were reviewed to evaluate the success rates of surgical techniques used to repair rectovaginal fistulae. A modified horizontal perineal technique was used in eight mares; it was unsuccessful in one mare, but the fistula was subsequently successfully repaired using the same technique. One rectovaginal fistula that was converted to a third-degree perineal laceration was successfully repaired on the third attempt, using the suture principles of the modified horizontal perineal technique. A modified transvaginal technique was used in three mares, in two of which it failed; their fistulae were subsequently converted to third-degree perineal lacerations and repaired using a one-stage closure. We conclude that the modified horizontal perineal technique is a one-stage procedure that is technically easy to perform, results in less tension on the suture lines, and is effective in modifying poor vulvar conformation. PMID- 17424201 TI - Neosporosis in a golden retriever dog from Ontario. PMID- 17424202 TI - Urethral recess dilatation in a Charolais-cross bull calf. PMID- 17424203 TI - Chronic mastitis caused by Haemophilus somnus in a dairy cow. PMID- 17424204 TI - Metastatic chondrosarcoma and subcutaneous granulomas in a grey rat snake (Elaphe obsoleta obsoleta). PMID- 17424205 TI - Early Canadian microscopists with associations to veterinary medicine. PMID- 17424206 TI - Ontario. Q fever abortions in a goat herd. PMID- 17424207 TI - Alberta. Porcine clostridial enteritis. PMID- 17424208 TI - British Columbia. Aspergillosis Outbreak in Stellar's jays (Cyanocitta stelleri) from central Vancouver Island. PMID- 17424209 TI - Mettre de l'ordre dans les ressources humaines. PMID- 17424210 TI - Les contrats d'emploi et les ententes restrictives. PMID- 17424211 TI - Veterinary Specialization - we already have it. PMID- 17424212 TI - Enforced preownership licensing will not lead to a more financially rewarding caseload. PMID- 17424213 TI - Relationship between high concentrate diets and treatment rates for respiratory infections in feedlot calves overstated. PMID- 17424214 TI - Extensive research needed to determine relationship between diet and respiratory infection in feedlot calves. PMID- 17424216 TI - Mycobacterium bovis in elk in Alberta. PMID- 17424215 TI - Anterior hormone secretion is episodic. PMID- 17424218 TI - Veterinary medical ethics. PMID- 17424217 TI - Disease-related stress affected pituitary-gonadal axis more than pituitary adrenal axis. PMID- 17424219 TI - Council agrees on new structure and vision for CVMA. PMID- 17424220 TI - Bureau of Veterinary Drugs: Suspected drug adverse reactions reported to The Bureau of Veterinary Drugs. PMID- 17424221 TI - Copper poisoning in a dairy herd fed a mineral supplement. AB - Copper poisoning in a dairy herd resulted in the death of 9 of 63 (14%) adult Holstein cows. Clinical signs were acute anorexia, weakness, mental dullness, poor pupillary light reflexes, and scant nasal discharge. These were followed by recumbency, chocolate-colored blood, jaundice, and death. Four animals exhibited signs of hyperesthesia and/or rumen stasis prior to death. At necropsy there was generalized icterus of body tissues, with the liver appearing orange and the kidneys dark blue. Histologically, there was accumulation of hemosiderin in Kupffer cells, and severe to moderate hepatocellular necrosis in all cases. Ammonium molybdate added to the ration, combined with the cessation of mineral supplementation, arrested the outbreak.These cases illustrate significant mortality, due to copper poisoning, in adult cattle fed a low-dose mineral dietary supplement for over two years. Dietary copper intake of the herd (on a dry matter basis) was 37.5 mg/kg for lactating cows and 22.6 mg/kg for dry cows. PMID- 17424222 TI - Evaluation of an in-office allergy screening test in nonatopic dogs having various intestinal parasites. AB - An in-office allergy screening kit was evaluated in 10 dogs that had no clinical signs of atopy but had various intestinal parasites. All 10 dogs had a positive reaction to the dust mite Dermatophagoides farinae, and four dogs had positive reactions to at least one other group of allergens. The kit as presently marketed gives numerous false positive results in parasitized nonatopic dogs. PMID- 17424223 TI - Clinical effects of detomidine with or without atropine used for arthrocentesis in horses. AB - The effectiveness of detomidine with or without atropine sulfate premedication in producing sedation and analgesia for arthrocentesis was studied in 12 horses. The effects were evaluated by monitoring heart and respiratory rates, borborygmi, distance from the lower lip to the floor, systolic blood pressure, and response to needle insertion. Either atropine or saline (as a placebo) was administered immediately prior to detomidine. All drugs were administered intravenously. Measurements were taken prior to drug injection and at 1, 5, 10, 15, 20, 25, 30, 40, 50, 60, 120, 180 and 240 minutes postinjection. Detomidine with atropine resulted in significantly higher heart rates than detomidine without atropine for the three hours of observation. Borborygmi were significantly decreased for four hours following detomidine with atropine and for three hours following detomidine without atropine, when compared to preinjection levels. Systolic blood pressure was significantly increased for 15 minutes following detomidine and atropine compared to the preinjection level. The head was markedly lowered for 60 minutes with either treatment. Atropine prevented the bradyarrhythmia and bradycardia induced by detomidine, but it induced a tachycardia. A satisfactory response for needle insertion and adequate synovial fluid aspiration was achieved in 95% of the trials with detomidine, with or without atropine sulfate premedication. The results suggest that, although atropine prevents bradyarrhythmia and bradycardia following detomidine, administering detomidine without atropine is satisfactory for arthrocentesis in untrained horses. PMID- 17424224 TI - Spironucleus-associated necrotic enteritis in angelfish (Pterophyllum scalare). PMID- 17424225 TI - Male pseudohermaphroditism of the testicular feminization type in a heifer. PMID- 17424226 TI - A retrospective study of frostbite injuries in calves (1982-1991). PMID- 17424227 TI - Alberta. Feline chlamydiosis in an animal shelter. PMID- 17424228 TI - Alberta. Tetanus in feeder calves associated with elastic castration. PMID- 17424229 TI - British Columbia. Infectious hematopoietic necrosis in Atlantic salmon in British Columbia. PMID- 17424230 TI - Massive mammary enlargement in a cat. PMID- 17424232 TI - Looking to the future. PMID- 17424233 TI - Dispenser beware. PMID- 17424235 TI - A year in review. PMID- 17424234 TI - Veterinary medical ethics. PMID- 17424236 TI - Auditor's report. PMID- 17424237 TI - Dr. Myron Mills - there's more than ET to this man. PMID- 17424238 TI - Massive bone tumor of the cranium in a dog. PMID- 17424239 TI - A field study on the usefulness of milk progesterone determination to confirm estrus and pregnancy of dairy cows in the Fraser Valley area of British Columbia. AB - A field study was conducted to determine the usefulness of milk progesterone determination at the time of breeding to confirm estrus and at 21 days postbreeding to detect open cows. Twenty-seven dairy farmers collaborated in this study by providing milk samples on the day of breeding and 21 days later, together with pregnancy diagnosis data and information on herd reproductive management. Herd size ranged from 15 to 175 cows, the average being 65 milking cows. Six hundred and sixty-seven breeding-day samples and 472, 21-day samples were provided by the farmers. Analysis of milk samples for progesterone by a solid phase radioimmunoassay showed that only 32 (4.8%) of the services were performed when the cow was not in estrus (progesterone > 1 ng/mL). Of the 472, 21 day samples, 337 (71%) showed progesterone levels of > 1 ng/mL, while 135 (29%) showed progesterone levels of < 1 ng/mL. Subsequently, 243 (72%) of the cows with progesterone > 1 ng/mL and eight (6%) of the cows with progesterone < 1 ng/mL were diagnosed pregnant by transrectal palpation, giving a pregnancy rate of 53%. Progesterone concentration on the day of breeding was not associated with season or herd size. However, progesterone concentration at 21 days and pregnancy rate were associated with herd size. These results indicate that fertillzation failure and/or early embryonic mortality, rather than inaccurate detection of estrus, are the major reproductive problems encountered by the dairy farmers in British Columbia. Furthermore, progesterone values at 21 days were closely related to reproductive status and indicate the usefulness of milk progesterone assay for the early detection of open cows. PMID- 17424240 TI - Newcastle disease in wild water birds in western Canada, 1990. AB - This report describes the investigation of mortality of double-crested cormorants (Phalacrocorax auritus), white pelicans (Pelecanus erythrorhynchos), and gulls (Larus spp.) in Alberta, Saskatchewan, and Manitoba during late summer 1990. Techniques used varied among areas, but virological and histopathological examination of birds was done in each area. The major clinical sign in cormorants was inability to fly, often with unilateral wing or leg paralysis. Focal nonsuppurative inflammation was present in the brain and spinal cord of cormorants and pelicans. Newcastle disease virus (NDV) was isolated from cormorants, a pelican, and a ring-billed gull (Larus delawarensls) from Saskatchewan. Cormorants from Alberta were positive for NDV in an immunofluorescent test. Most of the viruses were classed as velogenic and all had a similar monoclonal antibody profile to viruses from the 1970 to 1974 panzootic. Approximately half of cormorant, pelican, and gull eggs collected from affected colonies in the spring of 1991 had antibody to NDV. Antibody was also present in cormorant eggs from the Great Lakes. No unusual mortality was detected at any colony in 1991. Fledgling cormorants and gulls from colonies where mortality occurred in 1990 did not have antibody to NDV in June-July 1991. The overall extent of mortality among water birds and the source of the virus were not determined. PMID- 17424241 TI - Relationship of serum total calcium to serum albumin in dogs, cats, horses and cattle. AB - A retrospective study was performed in order to assess the relationship between serum calcium and serum albumin concentrations in domestic animals. Results of 9041 canine, 1564 feline, 2917 equine, and 613 bovine serum samples from hospitalized patients were examined by regression analysis. Subpopulations of cases with concurrent elevations in creatinine or that were less than six months of age were evaluated separately. Statistically significant linear relationships between calcium and albumin concentrations were established for each species (p <0.05). The coefficients of determination (r(2)) were 0.169 for dogs, 0.294 for cats, 0.222 for horses, and 0.032 for cattle. The correlation coefficients (r) computed were: dogs = 0.411, cats = 0.543, horses = 0.471, cattle = 0.182. Neither increases in creatinine concentration nor juvenile age appreciably influenced the relationship between calcium and albumin concentrations. Interspecies variation was marked, and a strong correlation between calcium and albumin concentrations was not established in any species. PMID- 17424242 TI - Pulsion diverticulum of the cranial cervical esophagus in a horse. PMID- 17424243 TI - Hyperkinesis in dogs: Six case reports. PMID- 17424244 TI - Immunotherapy of equine cutaneous lymphosarcome using low dose cyclophosphamide and autologous tumor cells infected with vaccinea virus. PMID- 17424245 TI - What is professional malpractice? PMID- 17424247 TI - Irregular approval of drug use by veterinary licensing bodies. PMID- 17424248 TI - Veterinary medical ethics. PMID- 17424249 TI - CVMA Insurance Survey: Insurance survey results. PMID- 17424250 TI - Evaluation of methods for dehydration of bovine colostrum for total replacement of normal colostrum in calves. AB - Different methods for the dry preservation of colostrum to be used as a total replacement for bovine colostrum were evaluated. Pooled colostrum from the first and second postpartum milkings from multiparous dairy cows containing immunoglobulin in excess of 40 g/L was freeze-dried, microwave vacuum evaporated, and spray-dried. Spray-drying produced a dried colostrum in which immunoglobulin quantity and function were preserved and was the most cost-effective. Other dehydration methods, although effectively conserving immunoglobulins, were too slow and costly to be used to produce a bovine colostrum replacer. Newborn, colostrum-deprived, dairy calves fed spray-dried colostrum containing 126 grams of immunoglobulin reconstituted in three liters of water as their sole source of immunoglobulin achieved normal mean serum immunoglobulin concentrations. Spray dried colostrum with high concentrations of immunoglobulin may be produced economically and used as an effective and convenient colostrum replacer in newborn calves. PMID- 17424251 TI - Saline drinking water in broiler and Leghorn chicks and the effect in broilers of increasing levels and age at time of exposure. AB - The purpose of this research was to assess the effect of increasing levels of salt and 5 g/L of salt (0.20% extra sodium) in the drinking water in broiler chickens at various ages up to five weeks of age and to compare the response of broiler chickens and White Leghorns to saline water (0.20% sodium). The effect was measured by the response of the right ventricle to pulmonary hypertension. The results indicated that broiler chickens under three weeks are more susceptible to saline water containing 0.20% sodium than those over three weeks of age, and that broilers given increasing levels of dietary salt may be more resistant to excess dietary salt than those that have had no previous exposure. The results also demonstrated that broiler chickens are more susceptible than White Leghorns to 0.20% extra dietary sodium in drinking water. We conclude that Leghorn chicks are more resistant to excess dietary sodium than broilers and that broilers become more resistant to saline water containing 0.20% sodium after three weeks of age. PMID- 17424252 TI - Magnetic resonance imaging of the canine and feline eye, orbit, and optic nerves and its clinical application. AB - The purpose of this study was to investigate magnetic resonance imaging of the normal canine and feline eye, orbit and optic nerves using proton density weighted, T(1)-weighted and T(2)-weighted images. The clinical application of magnetic resonance imaging in veterinary ophthalmology was also investigated using three clinical cases: a feline orbital melanoma, a feline optic nerve meningioma, and a canine orbital fibrosarcoma. Gadolinium diethylenetriamine pentaacetic acid enhanced magnetic resonance imaging was completed on the case of feline optic nerve meningioma. Magnetic resonance imaging provides excellent anatomical detail of the canine and feline eye, orbit, and optic nerves due to its superior soft tissue contrast, and its multiplanar and multislice imaging capability. Therefore it is of value for diagnostic imaging of some ophthalmic and neuro-ophthalmic conditions in the dog and cat. PMID- 17424253 TI - Metabolic acidosis without dehydration in a llama cria. PMID- 17424254 TI - Intoxication chronique au cuivre chez des agneaux lourds par l'eau d'abreuvement. PMID- 17424255 TI - In ever widening circles: Osler's influence on veterinary medicine in Sweden. PMID- 17424257 TI - Ontario. Winter death syndrome (exposure and starvation) in deer. PMID- 17424256 TI - Reactions medicamenteuses indesirables presumees qui ont ete signalees au Bureau des medicaments veterinaires. PMID- 17424258 TI - Quebec. Distribution of Streptococcus suis capsular types in Canada in 1992. PMID- 17424259 TI - Alberta. Polioencephalomalacia in a wild mule deer. PMID- 17424260 TI - Forelimb amputation in a red deer. PMID- 17424261 TI - Uterine prolapse in a camel. PMID- 17424262 TI - Forewarned is to be forearmed. PMID- 17424263 TI - Does veterinary dermatology interest you? PMID- 17424265 TI - Veterinary medical ethics. PMID- 17424264 TI - On ELISA for the determination of BHV-1 antibody. PMID- 17424266 TI - Is it really true? PMID- 17424267 TI - The quality of veterinary biological products: Behind the scenes. PMID- 17424268 TI - Importance of milk replacer intake and composition in rearing orphan foals. AB - Effects of milk replacer composition and intake on the growth of orphan foals were evaluated. Twenty foals were assigned to four treatments: 1) mare-nursed, 2) commercial foal milk replacer at recommended intakes (standard), 3) commercial foal milk replacer at high intakes (high), and 4) acidified replacer at recommended intakes (acidified). Foals fed milk replacer diets were weaned at 12 24 hours postpartum and fed milk replacer for 50 days. Mare-nursed foals were weaned between 52 and 56 days of age. Foals fed replacer diets gained 12% to 28% less weight than mare-nursed foals up to two weeks of age. However, by four months of age, weights of replacer-fed foals were similar to those of mare-nursed foals and 32 other mare-nursed foals at the farm weaned between three and four months postparium. Foals drank 10 to 12 L/100 kg body weight (BW) in fluid replacer daily over the trial period. During the first week, high intake foals consumed 26% more replacer (p<0.05) than foals fed acidified or standard diets. This higher intake resulted in diarrhea earlier (6-11 days vs 11-22 days) and for a longer time (6.3 days vs 2.5-3.6 days) than in foals fed recommended amounts. Mare-nursed foals developed "foal heat scours" in the second week postpartum. After the first week, foals fed high replacer diet voluntarily consumed the same volume of fluid replacer as foals fed the standard intake. Foals ate less than 1 kg grain mix/100 kg BW daily to one month of age, then increased intake to 1.5-2 kg/ 100 kg BW to weaning. Water intake was 20-40% of daily fluid intake and was correlated (r = 0.85) to dry matter intake. Foals in the high intake group ate less (p<0.05) solid feed and drank less water than foals fed the standard and acidified diets. The foal's stomach capacity appears to limit meal size and thus replacer intake. If recommended feeding intervals are used, replacer intakes by foals are less than 15% BW daily. High volume intakes appeared to prolong diarrhea. Normal growth rates occur when replacer and good-quality feeds are fed concurrently. PMID- 17424269 TI - An investigation of enzootic Glasser's disease in a specific-pathogen-free grower finisher facility using restriction endonuclease analysis. AB - Enzootic Glassers's disease was investigated to study the epidemiology of the disease strains on a farm where it presented a problem. Restriction endonuclease fingerprinting (REF) analysis technique was used, as all strains of Haemophilus parasuis are biochemically similar and many strains are biochemically untypable. After young weaned pigs were moved from farm A to farm B, Glasser's disease routinely occurred despite the use of antibiotics and a commercial bacterin. Isolates were taken from the nasal passages and from carcasses of clinically affected cases and subjected to REF analysis. Haemophilus parasuis was not isolated from any of the pigs on farm A, but it was isolated from 7/10 and 5/10 nasal swabs taken from farm B. Two H. parasuis strains isolated from clinical cases of Glasser's disease from farm B had an identical REF pattern, but were different from the nasal swabs and the H. parasuis strain contained in the bacterin. The subsequent use of a custom autogenous bacterin made from a clinical isolate of H. parasuis reduced the mortality rate on farm B. This investigation indicates that nasal isolates of H. parasuis are different than those causing clinical disease, and not all bacterin strains are cross protective for other strains. PMID- 17424270 TI - Bilateral neurogenic masseter muscle atrophy in a mare. PMID- 17424271 TI - Management of severe obstructive pulmonary disease with inhaled bronchodilator treatment in a horse. PMID- 17424272 TI - Congenital lordosis in three Haflinger foals. PMID- 17424273 TI - Surgical correction of medial luxation of the patella in a Miniature Horse. PMID- 17424275 TI - New Brunswick. Treatment of sea lice on Atlantic salmon with ivermectin. PMID- 17424274 TI - Isolation of Rhodococcus equi from the liver abscess of a goat in Trinidad. PMID- 17424276 TI - Ontario. Chromoblastomycosis in a horse. PMID- 17424277 TI - Alberta. Equine viral arteritis in a Thoroughbred filly. PMID- 17424280 TI - Veterinary medical ethics. PMID- 17424279 TI - Diagnostic ophthalmology. PMID- 17424282 TI - Manitoba veterinarian with special interest in animal welfare elected CVMA president. PMID- 17424281 TI - 1993 Association meetings in Edmonton. PMID- 17424283 TI - Veterinary continuing education and professional development: New concepts and solutions. PMID- 17424284 TI - Serological studies on the infection of dogs in Ontario with Borrelia burgdorferi, the etiological agent of Lyme disease. AB - A serological study was undertaken to determine whether dogs in Ontario are being exposed to Borrelia burgdorferi, the etiological agent of Lyme disease. This study consisted of a survey of randomly selected dogs and testing of diagnostic submissions from candidate Lyme disease cases. The survey of 1,095 dogs, bled between January 1988 and August 1989, revealed a total of 65 (5.9%) enzyme-linked immunosorbent assay (ELISA) reactors, of which 22 had immuno-fluorescent antibody assay (IFA) titers >/=1:32. All but one of the IFA-positive and 10 of the ELISA positive, IFA-negative sera were further tested by western blot. Eight western blot positive and three equivocal reactors were obtained. Three of the eight confirmed reactors had visited areas known to be endemic for Lyme disease, leaving five reactors that might have been infected in previously undocumented areas for B. burgdorferi activity in Ontario. Diagnostic submissions of sera from 223 dogs were received between August 1987 and February 1992. Test results revealed 21 (9.4%) IFA reactors, of which only six had significant titers (>/=1:256) and were reactive by an immunodot Borrelia test. All six dogs had travelled to known Lyme endemic areas. Based on results obtained from this study, it seems likely that the agent of Lyme disease is not widespread in Ontario. PMID- 17424285 TI - Zoonotic disease concerns in animal-assisted therapy and animal visitation programs. AB - A survey was done of 150 systematically selected United States animal care agencies and 74 Canadian humane societies to determine the prevalence of animal assisted therapy (AAT) programs; concerns about, and experience with, zoonotic diseases; and precautions taken to prevent zoonotic disease transmission. Of the 69 US agencies and 49 Canadian societies that reported having AAT programs, 94% used dogs and/or cats in their programs, 28% used rabbits, 15% used "pocket pets" (hamsters, gerbils, mice, guinea pigs), and 10% used birds (excluding poultry). About two-thirds of the programs were involved with the elderly in nursing homes, about a quarter of them worked with schools, and a quarter worked with hospitals. Half of the respondents had concerns about zoonotic disease control. Rabies, ringworm, and external parasitism were the most commonly cited zoonotic diseases of concern. Few concerns were based on actual experience. Fewer than half of the programs consulted a health professional about prevention of zoonotic diseases. Only 10% of the respondents reported having printed guidelines about the prevention of zoonotic disease transmission. Practising veterinarians are encouraged to make their expertise available to local AAT programs. PMID- 17424286 TI - Chromatolytic neuronal degeneration in a six-year-old cairn terrier. PMID- 17424288 TI - Cystic adenomatous hyperplasia of the cervical glands in a goat. PMID- 17424287 TI - Clinical and postmortem examination of sows culled for lameness. PMID- 17424290 TI - Preceptorship and the veterinary medical society of the ontario veterinary college. PMID- 17424289 TI - Uterine nocardiosis in two repeat breeder cows. PMID- 17424291 TI - Acetabular fracture repair. PMID- 17424292 TI - Practice tips to minimize exposure to litigation. PMID- 17424294 TI - Quebec. Isolation of Pasteurella caballi in a horse. PMID- 17424296 TI - Continued learning: you won't go far without it! PMID- 17424295 TI - Saskatchewan. Trichomonas fetus in bulls in Saskatchewan. PMID- 17424297 TI - Montreal Veterinary College class of 1879. PMID- 17424299 TI - Tails and specialties. PMID- 17424298 TI - Maternity leave; a legal right in Ontario. PMID- 17424300 TI - Ethical question concerning maternity leave poorly handled. PMID- 17424301 TI - Spread the word! PMID- 17424303 TI - Quiz corner. PMID- 17424302 TI - Veterinary medical ethics. PMID- 17424304 TI - Culture of bulk tank milk as a mastitis screening test: A brief review. AB - The culture of a sample of bulk tank milk may be a useful technique by which to screen herds for major mastitis pathogens. Staphylococcus aureus and Streptococcus agalactiae, if identified on a culture of a sample of bulk milk, reliably indicate infection of the udder. Environmental bacteria, such as the other streptococci and coliforms, are unlikely to be indicative of the proportion of cows infected with these organisms.Samples of bulk milk are readily obtainable and can be rapidly and inexpensively cultured to screen large numbers of herds for mastitis-causing bacteria, yet the performance of the test has only recently been formally assessed for its ability to correctly classify herds according to infection status.A single culture of bulk tank milk has been found to be a test with low sensitivity and high specificity for determining the presence of S. agalactiae or S. aureus in the herd. This means that many infected herds will be called negative, but few uninfected herds will be classified as positive.The literature assessing the performance of bulk tank milk culture in comparison with other mastitis screening tests, the use of bulk milk culture for prevalence surveys, and factors affecting these results is discussed. PMID- 17424305 TI - Evaluation of route and frequency of administration of three antimicrobial drugs in cattle. AB - This study in six cows compared serum concentrations of trimethoprim and sulphadoxine (16 mg/kg body weight (BW)) after once daily and twice daily administration, and of procaine penicillin G (20,000 IU/kg BW) after subcutaneous (SQ) and intramuscular (IM) administration, and evaluated postmortem tissue concentrations of penicillin following SQ treatment. Trimethoprim and penicillin were measured microbiologically, and sulphadoxine colorimetrically. Using minimum inhibitory concentrations (MIC), trimethoprim reached serum concentrations above 0.5 mug/mL from 15 minutes to 120 minutes, and sulphadoxine exceeded 9.5 mug/mL from 10 minutes to 12 hours, after administration. At 24 hours after treatment, both had declined to below the MIC of most organisms. A second treatment at 12 hours maintained concentrations of sulphadoxine above 9.5 mug/mL for a further 24 hours. For penicillin administered IM and SQ, concentrations that peaked at 0.88 mug/mL would inhibit most common grampositive bacteria for the entire 24 hour period and fastidious gram-negative organisms from 90 minutes to 12 hours after SQ treatment, but for virtually the entire period after IM administration. Mean +/- SD concentrations (mug/mL) of penicillin at euthanasia, five days after the last SQ administration, were 1.15 +/- 1.27 (injection site), 1.00 +/- 0.80 (liver), 0.90 +/- 0.58 (renal cortex), 0,58 +/- 0.17 (renal medulla), 0.13 +/- 0.11 (diaphragm), 0.10 +/- 0.08 (gluteal muscle), and 0.06 +/- 0.04 (fat). Therefore, except for the most sensitive organisms, twice daily injection of trimethoprim/sulphadoxine (16 mg/kg BW) may be required. Penicillin G administered SQ at 20,000 IU/kg BW should provide effective serum levels for as long as IM administration against gram-positive organisms, but for only about half as long against gram-negative bacteria. The label withdrawal time of five days cannot be used when penicillin is given SQ at 20,000 IU/kg BW for three days. PMID- 17424306 TI - Factors associated with past and potential future use of veterinarians by Ontario trout farmers. AB - This study investigated whether certain variables were associated with Ontario trout farmers' use of the Fish Pathology Laboratory (FPL), Ontario Veterinary College, or their willingness to pay for private veterinary services. Sixty-two farmers, representing 91% of Ontario's 1990 farmed trout production, were personally interviewed. Logistic regression was used to compare the 26 farmers who had submitted fish to the FPL during the previous two years to those who had not. Submitters tended to have been fish farming for fewer years, and to produce more fish, use more workers, treat prophylactically less frequently, keep records of drug treatments, and not disinfect holding units (p<0.05). Most farmers stated they would occasionally hire a private veterinarian. Only 13 farmers anticipated hiring one more than once per year. Polychotomous regression showed that farmers who purchased, instead of hatched, their seed stock, kept records, treated therapeutically often, and used several workers would hire private veterinarians more frequently (p<0.05). Strategies are discussed by which the veterinary profession might better understand and meet the needs of the aquaculture industry. PMID- 17424307 TI - A study of tilmicosin residues in milk following subcutaneous administration to lactating dairy cows. PMID- 17424308 TI - Interstitial pneumonia in piglets on Prince Edward Island. PMID- 17424309 TI - Osteosarcoma of the talus in a dog. PMID- 17424310 TI - The importance of accurate citrate to blood ratios in the collection of canine blood for hemostatic testing. PMID- 17424311 TI - Heartworm in dogs in Canada in 1991. AB - In late November 1991, 1883 clinics in Canada were sent a questionnaire to assess the status of Dirofilaria immitis in dogs in 1991 and there was a 60.0% response. There were 344,031 dogs tested for heart-worm (HW), 627 were found infected and the prevalence of HW infection was 0.18%. There were 417 dogs with HW in Ontario, 116 in Manitoba, 38 in Quebec, 53 in British Columbia, three in Alberta, and one in Nova Scotia. In British Columbia, all of the infected dogs but one were from the Okanagan valley which, as from 1991, is a new focus of infection in Canada. Most dogs with HW had not been on preventive medication in 1990, and the prevalence among dogs tested and unprotected was 0.59%. That prevalence was considerably higher in endemic areas. Companion dogs, over three years of age and maintained primarily outdoors in rural areas, were most frequently infected. One cat was diagnosed with D. immitis and 33 dogs had Dipetalonema reconditium. PMID- 17424313 TI - Alberta. Blindness in captive reared pheasants. PMID- 17424312 TI - Quebec. Actinobacillus suis in swine in southwestern Quebec. PMID- 17424314 TI - Diagnostic ophthalmology. PMID- 17424316 TI - The price of fame. PMID- 17424317 TI - Nonveterinarians providing "professional" services? PMID- 17424318 TI - Veterinary medical ethics. PMID- 17424319 TI - Quiz corner. PMID- 17424320 TI - Heifer ovariectomy using the Willis spay instrument: Technique, morbidity and mortality. AB - Two field trials were conducted to determine the morbidity and mortality rates in heifers ovariectomized with a Willis spay instrument.Trial A used 384 Hereford and Hereford cross, auction source, heifers, weighing an average of 230 kg. In trial B, 138 auction and ranch source heifers of mixed breeding and weighing an average of 285 kg were used. All heifers were ovariectomized per vaginam, with the ovaries left in the abdomen. Prophylactic antibiotics were administered and heifers were observed at pasture for the next 150 days.The morbidity and mortality rates for heifers in trial A were 0.5% and 0.26%, respectively. In trial B, there was no morbidity or mortality.These trials indicate that the technique described is a low risk, expedient method of ovariectomizing heifers. PMID- 17424321 TI - A field investigation of the economic impact of respiratory disease in feedlot calves. AB - A trial involving 512 beef calves was conducted in a commercial research feedlot to determine the effect of bovine respiratory disease (BRD) on performance parameters and carcass characteristics. Two hundred and fifty-six calves that were deemed to be "sick" (S) from BRD were allocated to 16 pens and 256 calves that were considered to be "well" (W) were allocated to another 16 pens. The outcome variables that were measured included average daily gain (ADG), daily dry matter intake (DDMI), dry matter intake to gain ratio (DM:G), BRD treatment rate, death loss, carcass traits, and net profit per pen.The data were partitioned into several time intervals including processing (P) to day -1, day 0 to day 27, day 28 to day 55, day 56 to day 83, day 84 to day 111, day 112 to day 139, day 140 to slaughter, day 0 to slaughter (0-Slaugh), and processing to slaughter (P-Slaugh). However, the most important interval was from processing to slaughter.For the interval P-Slaugh, there were no significant (p>/=0.05) differences between the S and W groups with respect to ADG and DM:G. Also, for the interval 0-Slaugh, the DDMI was similar for both groups. There were no significant (p>/=0.05) differences between the S and W groups for carcass weight, average fat, grade fat, rib eye area, marbling score, cutability estimate, or carcass grade distribution.The BRD treatment rates in the S and W groups were 6.6% and 4.7%, respectively. The mortality rates in the S and W groups were 0.78% and 0.39%, respectively. Also, there were no deaths attributable to BRD in either group.In the economic model, there was no significant (p>/=0.05) difference between the S and W groups with respect to net profit per pen.We conclude that this trial did not validate the concept that BRD impacts performance parameters, because a sufficient disease challenge was not present. However, this study provides several observations that will enhance the experimental design of future studies that attempt to quantify the total economic impact of BRD. PMID- 17424322 TI - Xylazine epidural analgesia for cesarian section in cattle. PMID- 17424323 TI - Infective endocarditis in a dairy farmer in association with bovine Staphylococcus aureus mastitis. PMID- 17424324 TI - Type III von Willebrand's disease in Scottish terriers: A report of two cases. PMID- 17424325 TI - Acromegaly in a cat: Diagnosis by magnetic resonance imaging and treatment by cryohypophysectomy. PMID- 17424326 TI - Repair of fractures of the radius and ulna in a ewe using positive profile transfixation pins and casting. PMID- 17424327 TI - Wounded in action: Sergeant Billy, the goat in military service for Canada in the Great War. PMID- 17424328 TI - Floating a loan. PMID- 17424331 TI - Time to look in both directions. PMID- 17424329 TI - Sudden death of a heifer caused by bleeding form the reticular vein. PMID- 17424332 TI - Wildlife health and welfare. PMID- 17424333 TI - Uterine nocardiosis not necessarily the sole cause of repeat breeding. PMID- 17424335 TI - Veterinary medical ethics. PMID- 17424334 TI - Nocardia asteroides in the uterus of two repeat breeder cows. PMID- 17424336 TI - Quiz corner. PMID- 17424337 TI - The effects of spaying and anabolic implants on the average daily weight gain of heifers on pasture. AB - Two studies were carried out to determine the effects of spaying and implanting on the growth of heifers and compare Ralgro and Synovex-S for spayed heifers. In the first study, 121 crossbred heifers were randomly allocated to four groups: intact and no implant; intact and progesterone and estradiol benzoate (Synovex S); spayed and no implant; and spayed and Synovex-S. The average daily gain (ADG) and body weight (BW) data were analyzed as a 2 x 2 factorial experiment with a pasture effect which was considered as a block. Based on ADG for days 0-120, the response to implanting was 17.6% for spayed heifers and 1.8% for intact heifers. In the second study, 31 spayed heifers implanted with zeranol (Ralgro) were compared with 30 spayed heifers implanted with Synovex-S. The data were analyzed as a one-way analysis of variance. The ADG for spayed heifers on Ralgro did not differ significantly (p>0.05) from those implanted with Synovex-S over a period of 92 days, suggesting that both implants produce similar growth responses. PMID- 17424338 TI - The effect of route and dosage of immunization on the serological response to a Pasteurella haemolytica and Haemophilus somnus vaccine in feedlot calves. AB - The effect of route and dosage of administration on the serological response to a vaccine containing genetically attenuated leukotoxin of Pasteurella haemolytica combined with bacterial extracts of P. haemolytica and Haemophilus somnus (Somnu Star Ph, Biostar Inc., Saskatoon, Saskatchewan) was evaluated in a controlled field trial in 301 feedlot calves. Vaccination of calves on arrival at the feedlot with Somnu-Star Ph significantly (p < 0.05) increased P. haemolytica and H. somnus serum antibody titers and reduced bovine respiratory disease (BRD) morbidity. A single subcutaneous vaccination with Somnu-Star Ph was as effective in stimulating a humoral antibody response and in reducing BRD morbidity as double vaccination by the intramuscular or the subcutaneous route. Furthermore, there were no swellings or adverse reactions observed with either subcutaneous or intramuscular administration of Somnu-Star Ph.These results suggest that feedlot calves can be immunized subcutaneously once on arrival with Somnu-Star Ph. Double vaccination was of no added value in this trial, because the majority of BRD morbidity occurred prior to revaccination fourteen days postarrival. Additional larger-sized field trials are needed to monitor the duration of immunity following vaccination and to test the effect of route and dosage of vaccination on mortality. PMID- 17424339 TI - Comparison of two formulations of oxytetracycline given prophylactically to reduce the incidence of bovine respiratory disease in feedlot calves. AB - A trial involving 1,803 feedlot calves was conducted under commercial feedlot conditions in western Canada to compare the relative effectiveness of a new oxytetracycline formulation, administered either intramuscularly (BMI) or subcutaneously (BMS), to a currently available oxytetracycline formulation, administered intramuscularly (LAB), for the prevention of bovine respiratory disease (BRD) in feedlot calves. All experimental treatments were administered upon arrival at the feedlot and again on the third day after arrival.Over the entire feeding period, there were no significant differences (p>/=0.05) in the BRD treatment rates or the BRD relapse rates between either the BMI or BMS groups compared to the LAB group. Similarly, there were no significant differences (p>/=0.05) in the BRD treatment rates in the BMI or BMS groups from days 8-14, days 15-90, or days 1-90 of the feeding period compared to the LAB group. However, during the first seven days of the feeding period the BRD treatment rate in the BMI group was 1.55 times (p<0.05) higher than in the LAB group. From days 1-90 and day 1 to the end of the feeding period, the overall mortality rates, BRD mortality rates, and BRD case fatality rates were two to six times lower in the BMS and BMI groups as compared to the LAB group; however, these differences were not statistically significant (p>/=0.05).These data indicate that both the intramuscular and subcutaneous administration of a new oxytetracycline formulation are comparable to the intramuscular administration of a currently available oxytetracycline formulation when given to calves upon arrival at the feedlot. PMID- 17424340 TI - Periosteal osteosarcoma of the scapula in a horse. PMID- 17424341 TI - Rickettsial septicemia in farmed Atlantic and chinook salmon in British Columbia: Clinical presentation and experimental transmission. PMID- 17424342 TI - Prince Edward Island. Drug resistance patterns among Salmonella isolates. PMID- 17424343 TI - British Columbia. Avian tuberculosis in ostriches. PMID- 17424344 TI - Directors' liability for environmental matters. PMID- 17424346 TI - Council takes action on priorities. PMID- 17424345 TI - Diagnostic ophthalmology. PMID- 17424347 TI - Quiz corner. PMID- 17424348 TI - The use of antimicrobial drugs in cattle. PMID- 17424349 TI - The fallen world. PMID- 17424351 TI - Quiz corner. PMID- 17424350 TI - Pour se lancer - sans tomber - dans un emprunt. PMID- 17424352 TI - Canadian condemnation rate for bovine ocular squamous cell carcinoma. PMID- 17424353 TI - Quiz corner. PMID- 17424354 TI - Role of the CVMA in international veterinary medicine. PMID- 17424355 TI - First confirmed case in Alberta of eperythrozoonosis in swine. PMID- 17424357 TI - Quiz corner. PMID- 17424356 TI - Cvma-auditor's report. PMID- 17424358 TI - Links with the past. PMID- 17424359 TI - Veterinary medicine cum law. PMID- 17424360 TI - Quiz corner. PMID- 17424361 TI - British Columbia. Acute lead poisoning in a bald eagle secondary to bullet ingestion. PMID- 17424362 TI - The passing of a friend 1911-1994. PMID- 17424363 TI - Canadian veterinarians contribute to veterinary medicine in Latvia. PMID- 17424365 TI - Quiz corner. PMID- 17424364 TI - Christian veterinary missions of Canada. PMID- 17424366 TI - The year in review. PMID- 17424367 TI - Intellectual property rights and private practice - A reply. PMID- 17424368 TI - Quiz corner. PMID- 17424370 TI - Clenbuterol not approved for use in cattle in Canada - A reply. PMID- 17424369 TI - International veterinary nurses and technicians association world congress - England 1993. PMID- 17424371 TI - Quiz corner. PMID- 17424372 TI - Commentary on the results of the economic survey of veterinary practice in Atlantic Canada - A reply. PMID- 17424373 TI - Commentary on the results of the economic survey of veterinary practice in Atlantic Canada. PMID- 17424374 TI - Quiz corner. PMID- 17424375 TI - SPECIAL FEATURE: Gorging on nouns: Why scientists write as they do. PMID- 17424376 TI - Quiz corner. PMID- 17424379 TI - Quiz corner. PMID- 17424377 TI - Primaute: le public ou la profession? Propriete en commun et propriete non veterinaire d'une pratique veterinaire. PMID- 17424382 TI - Quiz corner. PMID- 17424381 TI - The end of an era. PMID- 17424383 TI - Quiz corner. PMID- 17424384 TI - Mucosal disease and acute bovine diarrhea go together - A reply. PMID- 17424385 TI - Animal abuse. PMID- 17424386 TI - AABP acts to attract veterinary graduates to food animal practice. PMID- 17424387 TI - We must do our homework. PMID- 17424388 TI - Veterinary medical imaging: Other considerations in critical thinking - A reply. PMID- 17424389 TI - Quiz corner. PMID- 17424390 TI - What is this stuff called Petstuff? PMID- 17424392 TI - Quiz corner. PMID- 17424391 TI - Shortage of rural, mixed animal practitioners. PMID- 17424394 TI - Quiz corner. PMID- 17424393 TI - Heavy metal toxicosis and slaughter for human consumption - A reply. PMID- 17424396 TI - Sociological backgrounds of students at the Western College of Veterinary Medicine. PMID- 17424395 TI - Caring for pets of immunocompromised persons. PMID- 17424398 TI - Selection of students for veterinary medicine - A reply. PMID- 17424397 TI - The future of agricultural veterinary practice. PMID- 17424399 TI - Quiz corner. PMID- 17424400 TI - Shortage of rural, mixed animal practitioner - A reply to the replies. PMID- 17424401 TI - Quiz corner. PMID- 17424403 TI - Quiz corner. PMID- 17424402 TI - Evaluation of methods for the euthanasia of cattle in a foreign disease outbreak A reply. PMID- 17424405 TI - Calling for articles on small companion animals. PMID- 17424404 TI - Le personnel. PMID- 17424406 TI - Quiz corner. PMID- 17424407 TI - Quiz corner. PMID- 17424409 TI - Commit to publish. PMID- 17424411 TI - Quiz corner. PMID- 17424410 TI - Bovine somatotropin and the regulatory process for veterinary drug approval - A reply. PMID- 17424413 TI - Quiz corner. PMID- 17424412 TI - Holistic medicine not an alternative therapy. PMID- 17424415 TI - Quiz corner. PMID- 17424417 TI - Quiz corner. PMID- 17424416 TI - Something to chew on - Erratum. PMID- 17424418 TI - Prescription of medications that are neither therapeutic nor prophylactic to increase productivity - A reply. PMID- 17424419 TI - Dry sow stalls - an animal welfare issue: A reply. PMID- 17424420 TI - Quiz corner. PMID- 17424421 TI - Quiz corner. PMID- 17424423 TI - Assistance in dealing with pet loss bereavement in children. PMID- 17424424 TI - Quiz corner. PMID- 17424425 TI - Quiz corner. PMID- 17424427 TI - Quiz corner. PMID- 17424426 TI - The universality of CCAC's program threatened. PMID- 17424429 TI - A tribute to Dr. Robert A. Curtis: Teacher, scholar, and clinician. PMID- 17424430 TI - Quiz corner. PMID- 17424431 TI - Is there copyright on the Internet? PMID- 17424432 TI - Case for negligence requires closer examination - Replies. PMID- 17424433 TI - Case for negligence requires closer examination - Replies. PMID- 17424434 TI - Cvma annual report. PMID- 17424435 TI - Quiz corner. PMID- 17424436 TI - Raccoon rabies in eastern Canada (1963 to 1994): A retrospective report. PMID- 17424437 TI - Quiz corner. PMID- 17424438 TI - Dr. Robert A. Curtis: An appreciation. PMID- 17424440 TI - Hodgepodge. PMID- 17424439 TI - Occupational health and safety quiz. PMID- 17424441 TI - Pro canine breed specific legislation - A reply. PMID- 17424443 TI - Information by prescription: client information handouts for the practitioner exotics-reptiles; feline; canine; pet birds. PMID- 17424442 TI - Quiz corner. PMID- 17424445 TI - Quiz corner. PMID- 17424444 TI - Diseases of calves. PMID- 17424446 TI - Prescribing and selling pharmaceuticals. PMID- 17424447 TI - Responsibility for returning lost property. PMID- 17424449 TI - Occupational health and safety quiz. PMID- 17424450 TI - Nonsteroidal anti-inflammatory analgesics in pain management in dogs and cats - A correction. PMID- 17424451 TI - Rabbit news and research. PMID- 17424452 TI - New veterinary website. PMID- 17424453 TI - Quiz corner. PMID- 17424455 TI - Employment trends. PMID- 17424456 TI - Inappropriate comment. PMID- 17424457 TI - Quiz corner. PMID- 17424458 TI - NEWFOUNDLAND AND LABRADOR. Rabies in Labrador. PMID- 17424459 TI - QUEBEC. Urinary tract infection associated with Corynebacterium pilosum in a dog. PMID- 17424460 TI - Extra label drug use - A reply. PMID- 17424463 TI - Inappropriate title. PMID- 17424461 TI - Occupational health and safety quiz. PMID- 17424464 TI - Using "data" correctly. PMID- 17424465 TI - Editor's comment. PMID- 17424466 TI - Cvma annual report. PMID- 17424467 TI - Quiz corner. PMID- 17424468 TI - Veterinary medical ethics. PMID- 17424469 TI - Quiz corner. PMID- 17424471 TI - What price experience? PMID- 17424472 TI - Occupational health and safety quiz. PMID- 17424473 TI - Incorrect answer. PMID- 17424474 TI - More incorrect word usage. PMID- 17424475 TI - Pet owners assume competency when selecting their veterinarian. PMID- 17424476 TI - Pet owners assume competency when selecting their veterinarian - A reply. PMID- 17424477 TI - Quiz corner. PMID- 17424478 TI - Over regulation - Enough already! PMID- 17424479 TI - Streaming and licensure - A reply. PMID- 17424480 TI - Quiz corner. PMID- 17424481 TI - Ethics on the decline - A reply. PMID- 17424482 TI - Ethics on the decline. PMID- 17424483 TI - Working with the financially embarrassed client. PMID- 17424484 TI - Veterinarian's responsibility in regard to a stray tattooed pet. PMID- 17424485 TI - Foreign locums the answer to shortage of large animal practitioners. PMID- 17424487 TI - Occupational health and safety quiz. PMID- 17424486 TI - Foreign locums the answer to shortage of large animal practitioners - A reply. PMID- 17424489 TI - Editor's note. PMID- 17424490 TI - Quiz corner. PMID- 17424491 TI - Over regulation - Lawyers not the culprits. PMID- 17424493 TI - Ask not. PMID- 17424492 TI - Using the fentanyl patch - A reply. PMID- 17424494 TI - Comments regarding the treatment of lead toxicosis in rabbits-A reply. PMID- 17424495 TI - Occupational health and safety quiz. PMID- 17424496 TI - Use of rabbits in ostrich rearing. PMID- 17424498 TI - Editor's comment. PMID- 17424497 TI - Wanted, a toll-free number for advice on suspect cases of foreign animal disease A reply. PMID- 17424500 TI - Editor's comment. PMID- 17424499 TI - Dogtrek 97-98. PMID- 17424501 TI - A welfare concern. PMID- 17424502 TI - Veterinary medical ethics. PMID- 17424503 TI - Quiz corner. PMID- 17424506 TI - Pediatric neuters can be technically challenging. PMID- 17424505 TI - Quiz corner. PMID- 17424507 TI - Strong veterinary leadership required for the 21st century. PMID- 17424508 TI - Photocopying a no-no. PMID- 17424509 TI - Quiz corner. PMID- 17424510 TI - Gum boots for horses. PMID- 17424512 TI - Questioning some of the CVMA pet food recommendations - A reply. PMID- 17424513 TI - Prohibition of clenbuterol use in food-producing animals - A reply. PMID- 17424514 TI - Prohibition of clenbuterol use in food-producing animals. PMID- 17424515 TI - In support of friendly practice guidelines. PMID- 17424516 TI - Occupational health and safety quiz. PMID- 17424517 TI - Diagnostic application of avian endoscopy. PMID- 17424518 TI - Quiz corner. PMID- 17424519 TI - Bladder lavage for rewarming a hypothermic dog. PMID- 17424521 TI - Why not? PMID- 17424522 TI - Hear all ye dairy cattle veterinarians. PMID- 17424523 TI - Quiz corner. PMID- 17424524 TI - Friendly practice guidelines problematic - A reply. PMID- 17424525 TI - Medical Research Council launches new health research funding initiative - Canadian veterinarians urged to join the colleges in lobbying for support for veterinary research. PMID- 17424526 TI - Occupational health and safety quiz. PMID- 17424527 TI - A new Journal feature - Student Papers. PMID- 17424528 TI - Quiz corner. PMID- 17424530 TI - Veterinary medical ethics. PMID- 17424529 TI - New Canadian veterinary acupuncture organization. PMID- 17424532 TI - The importance of Correct punctuation. PMID- 17424531 TI - Occupational health and safety quiz. PMID- 17424534 TI - Financing veterinary education and research: The challenge facing the profession. PMID- 17424535 TI - Apropos the 1998 Hill's Public Relations Award. PMID- 17424536 TI - Concern about the possibility of selenium in the food chain - A reply. PMID- 17424537 TI - Quiz corner. PMID- 17424538 TI - Veterinary research and human health. PMID- 17424539 TI - Origines de la surveillance infranationale au Canada. PMID- 17424540 TI - Demographic analysis of the veterinary profession in Canada - A reply. PMID- 17424541 TI - Concerns about Canada Post's stamp issue saluting the circus tradition. PMID- 17424542 TI - Concerns about Canada Post's stamp issue saluting the circus tradition. PMID- 17424543 TI - Quiz corner. PMID- 17424544 TI - Rabbit news and research quarterly. PMID- 17424545 TI - Calling all Canadian veterinary orthopedic surgeons. PMID- 17424546 TI - Deans respond to Task Force report "Opportunity for Renewal": A reply. PMID- 17424547 TI - Dr. Dan unruh. PMID- 17424548 TI - Pet loss support. PMID- 17424549 TI - Environmental concerns regarding the use of the ivermectin sustained-release bolus in cattle - A reply. PMID- 17424550 TI - Quiz corner. PMID- 17424551 TI - Membership in the CVMA. PMID- 17424552 TI - The CAHI versus the BVD - A reply. PMID- 17424553 TI - Membership in the CVMA-A reply. PMID- 17424554 TI - Quiz corner. PMID- 17424556 TI - Quiz corner. PMID- 17424555 TI - Erroneous statements in veterinary medical ethics' columns - A reply. PMID- 17424557 TI - Putting the ;I's back into ;Team'. PMID- 17424559 TI - Information requested from Jewish veterinarians. PMID- 17424558 TI - Pesticides must be sold in their original packaging. PMID- 17424560 TI - Inappropriate comment regarding fees charged by neighboring colleagues - A reply. PMID- 17424561 TI - Inappropriate comment regarding fees charged by neighboring colleagues - A reply. PMID- 17424562 TI - Veterinary medical ethics. PMID- 17424563 TI - Quiz corner. PMID- 17424564 TI - Veterinary education: time to abandon the cult of coverage. PMID- 17424566 TI - Veterinary medical ethics. PMID- 17424565 TI - Intubation not for guinea pigs - A reply. PMID- 17424567 TI - Cvma annual report. PMID- 17424568 TI - Quiz corner. PMID- 17424569 TI - CVMA insurance program. PMID- 17424570 TI - Quiz corner. PMID- 17424571 TI - ONTARIO. Raccoon rabies in eastern Ontario. PMID- 17424572 TI - Caveat: Explain the variables affecting the effectiveness of homeopathic therapy. PMID- 17424573 TI - Merging is the only way. PMID- 17424574 TI - Confusing ethics and religion - A reply. PMID- 17424576 TI - Quiz corner. PMID- 17424575 TI - Confusing ethics and religion. PMID- 17424577 TI - CVMA wrong to all itself with PetCare Insurance Brokers Ltd. - A reply. PMID- 17424578 TI - CVMA wrong to ally itself with PetCare Insurance Brokers Ltd. PMID- 17424579 TI - Thoughts for a friend. PMID- 17424580 TI - Acupuncture founded on magical and mystical musings about how the world works - A reply. PMID- 17424581 TI - Veterinary volunteers at the Pan American Games 1999. PMID- 17424582 TI - Quiz corner. PMID- 17424583 TI - Remuneration. PMID- 17424584 TI - Lifestyles. PMID- 17424585 TI - Implementation. PMID- 17424586 TI - Selection. PMID- 17424587 TI - Education. PMID- 17424588 TI - Veterinary homeopathy - A rebuttal - A reply. PMID- 17424590 TI - Points on acupuncture. PMID- 17424589 TI - Quiz corner. PMID- 17424591 TI - Comment regarding revaccination guidelines - A reply. PMID- 17424593 TI - Quiz. PMID- 17424592 TI - Claims for veterinary chiropractic unjustified - A reply. PMID- 17424594 TI - The Atlantic Veterinary College: where technology meets the sea. PMID- 17424596 TI - Quiz corner. PMID- 17424595 TI - Work force oversupply not a problem - A reply. PMID- 17424598 TI - Quiz corner. PMID- 17424597 TI - Simple facts and numbers - A reply. PMID- 17424600 TI - Comments on anesthesia and analgesia - A reply. PMID- 17424599 TI - Comments regarding the claimed effectiveness of bovine colostrum - A reply. PMID- 17424601 TI - Quiz corner. PMID- 17424603 TI - A small international effort - A big personal reward. PMID- 17424602 TI - Another 2-way MLV vaccine against agents of bovine respiratory disease - A reply. PMID- 17424604 TI - Quiz corner. PMID- 17424605 TI - "Microchipping" - A technology in transition. PMID- 17424607 TI - Quiz corner. PMID- 17424606 TI - Should the convention program be modified? PMID- 17424608 TI - Shouldn't Canadian veterinary colleges be providing training in veterinary dentistry? - A reply. PMID- 17424609 TI - Vaccine against agents of bovine respiratory disease - A correction. PMID- 17424610 TI - Quiz corner. PMID- 17424611 TI - Quiz corner. PMID- 17424612 TI - ONTARIO. Hepatic lipodystrophy in 2 Galloway calves. PMID- 17424614 TI - Quiz corner. PMID- 17424613 TI - ONTARIO. Congenital brain edema in 2 Hereford calves. PMID- 17424616 TI - Quiz corner. PMID- 17424615 TI - The interpretation of "scientific" - A reply. PMID- 17424617 TI - Veterinary medical ethics. PMID- 17424618 TI - Quiz corner. PMID- 17424619 TI - Pet Food Certification Program survey. PMID- 17424620 TI - Dr. Mark lawrence. PMID- 17424621 TI - Pet Food Certification Program survey - A reply. PMID- 17424622 TI - Quiz corner. PMID- 17424623 TI - Are Web sites for veterinary practices worthwhile? PMID- 17424624 TI - Quiz corner. PMID- 17424625 TI - Whither the veterinary medical profession in the 21st century? PMID- 17424626 TI - Biting Dogs and kicking horses. PMID- 17424627 TI - Quiz corner. PMID- 17424628 TI - BARF: "Bones and Raw Foods" or "Biologically Appropriate Raw Foods" - Fad, fiction or fanatical? - A reply. PMID- 17424629 TI - CVMA - Decoding Success "An Inside Look at Canada's Top-Performing Practices". PMID- 17424631 TI - Veterinary medical ethics. PMID- 17424630 TI - CVMA - Decoding Success "An Inside Look at Canada's Top-Performing Practices" - A reply. PMID- 17424632 TI - Quiz corner. PMID- 17424633 TI - Questions associated with proposed changes to the Criminal Code: The CVMA Animal Welfare Committee responds. PMID- 17424634 TI - Large animal practitioners in Ontario deserve better support from their veterinary association(s). A reply from the Ontario Veterinary Medical Association. PMID- 17424636 TI - Quiz corner. PMID- 17424635 TI - Large animal practitioners in Ontario deserve better support from their veterinary association(s). A reply from the College of Veterinarians of Ontario. PMID- 17424638 TI - Concern regarding the News article - CVMA answers the call. PMID- 17424639 TI - Concern regarding the News article - CVMA answers the call - A reply. PMID- 17424640 TI - Quiz corner. PMID- 17424641 TI - Further thoughts on whether the veterinary profession is losing its way. PMID- 17424643 TI - Electronic archiving - Planning another service for CVMA members and journal subscribers. PMID- 17424644 TI - Canadian veterinarians lead the way. PMID- 17424645 TI - Are my CVMA membership dues being well spent? PMID- 17424647 TI - Quiz corner. PMID- 17424646 TI - Are my CVMA membership dues being well spent? - A reply. PMID- 17424648 TI - Evaluating your employees - Who, what to use, when, and how to deal with this sensitive topic to achieve results. PMID- 17424650 TI - Questions regarding suspected copper toxicity in a 10-year-old alpaca - A reply. PMID- 17424649 TI - Praise for the Commentary on structuring the veterinary curriculum and profession to promote the diversity of the profession. PMID- 17424651 TI - Quiz corner. PMID- 17424652 TI - Quiz corner. PMID- 17424653 TI - Proteins in human milk. AB - The human baby is born extremely immature, with its major organs and immune system not fully developed. for its survival, the infant depends on an extraordinarily well-adapted evolutionary strategy shared by all mammals: breastfeeding. But what does milk contain that makes it so essential for the newborn and how does it provide immunity, nutrition, and a source for optimal growth? Human milk is a very complex living fluid which comprises proteins, carbohydrates, lipids, cells and other biologically important components. These milk components interact synergistically with each other and their environment (the infant's gut) at a biomolecular level with the final result being that breastmilk feeds and protects the newborn. This article summarises the key characteristics of breastmilk proteins and describes their functions as critical molecules conferring human milk with its diverse bioactive properties. Also presented are some of the factors which hav an influence on the quantity and quality of breastmilk proteins. PMID- 17424654 TI - Ethical responsibilities of the Australian media in the representations of infant feeding. AB - Despite the fact that human milk is the ideal food for human babies, many Australian babies are still weaned sooner than the World Health Organization guidelines recommend. Australian mothers want to breastfeed--and initially do so. However, there is a rapid decline in the percentage of babies being offered breastmilk as newborns compared to six months old. Data collected in 2004 indicated that although 90% of newborn infants in NSW were put to the breast, or offered expressed milk, at least once, only 77% of infants were receiving at least some breastmilk regularly at the end of their first month. By six months of age only 4.6% of babies in NSW were being exclusively breastfed. This paper aims to analyse some of the reasons women in Australia prematurely wean their infants. Particular emphasis will be given to the representation of infant feeding in the media, how consumers use this information to make decisions about infant feeding, and the ethical responsibilities of said media in their portrayal of infant feeding. PMID- 17424655 TI - The contribution of infant food marketing to the obesogenic environment in Australia. AB - Obesity has been growing rapidly among both children and adult Australians in recent decades, raising concern at the associated chronic disease burden, and generating debate over the extent of individual versus government responsibility. This paper briefly reviews recent scientific evidence on links between poor early life nutrition and obesity in later life, which suggests that artificial baby milk rather than breastfeeding in infancy is associated with a 30-50% higher likelihood of later life obesity. It then presents data on long-term trends in breastfeeding in Australia and on consumption of infant milk products since 1939. Evidence is also presented of increased marketing and promotion of breastmilk substitutes from the mid 1950s, including through the healthcare system, associated with the emergence of increased competition in the Australian infant food industry. This collaborative marketing effort by industry and health professionals in turn contributed importantly to the sharp decline in breastfeeding from the mid 1950s. As a consequence, most Australians born since 1955 were exposed to artificial baby milk in early infancy. A substantial proportion of Australian infants are still partially fed with artificial baby milk in the first 12 months of life. The example of infant food highlights that the healthcare system and the food industry, and not just individual mothers' choices, have contributed to poor infant nutrition and obesity trends in Australia. Redressing healthcare system and industry practices to restore a supportive environment for breastfeeding is thus argued to be a necessary element of the public health response to the current obesity problem. PMID- 17424656 TI - Gastrointestinal endoscopy. PMID- 17424657 TI - Emergency ultrasound. PMID- 17424658 TI - Whiplash injuries can be visible by functional magnetic resonance imaging. PMID- 17424659 TI - Whiplash injuries can be visible by functional magnetic resonance imaging. PMID- 17424660 TI - Clinical ethics: a practical approach. PMID- 17424661 TI - [Rural family and "integrated system": a model of social humanism, 1880-1910]. PMID- 17424662 TI - [(Re)constructions of images of the couple in the Iberian Peninsula in the 16th and 17th centuries]. PMID- 17424663 TI - [The territorial origins and attitudes toward death in the Havana squadron, 1780 83]. PMID- 17424664 TI - [A reading of the long period: Spain from the 16th to the 18th century]. PMID- 17424665 TI - ["Amaos...y no os multipliqueis": ethical-sexual discourse of individualistic anarchism in Catalonia, 1927-37]. PMID- 17424666 TI - The legacy of Tycho Brahe. PMID- 17424667 TI - The influence of the wider kin group on individual life-course transitions: results from the Pays de Herve (Belgium), 1846-1900. PMID- 17424668 TI - Body and enlightenment in Denmark: on Matthias Lunding's diary 1787 and its context. PMID- 17424669 TI - "Be fruitful and multiply": genesis and generation in reformation Germany. PMID- 17424670 TI - German intellectual history and American romanticism: spirit, nature philosophy, and the occult. PMID- 17424671 TI - [Hygiene of the imagination, interpretation of script: juricial trends of dream and imagination]. PMID- 17424672 TI - [CME-ultrasonography 15/ Resolve]. PMID- 17424673 TI - Type 2 diabetes mellitus: magnitude of the problem and failure to achieve glycemic control. PMID- 17424674 TI - Islet dysfunction: an underlying defect in the pathophysiology of type 2 diabetes. PMID- 17424675 TI - Incretin-based therapies for type 2 diabetes: clinical utility. PMID- 17424676 TI - [Surgical pleural biopsy]. AB - The authors emphasize on the importance of histologic study of the sick pleura to obtain a diagnosis and to decide the treatment, operative or not, which must be specific. They choose the surgical biopsy and analyze the results obtained in 104 operations performed on 100 patients. Four groups of lesions are established: 1) malignant tumors (primary and secondary), 2) Non specific pleuritis consecutive to carcinoma of the lung or other organs, usually metastatic from the breast or the digestive tract, 3) Non specific pleuritis, and 4) Tuberculosis. Cancer or tuberculosis are found in pleurisies of different types. Cancer was found in a 43,7% of the serous pleurisies, in a 64% of the seroushemorrhagic, and in a 46% of the hemorrhagic pleurisies. Tuberculosis was found in a 15,6 % of the serous, in a 7,14 % of the seroushemorrhagic and in a 15,7 % of the purulent. In one case of hemorrhagic pleurisy in which the diagnosis of tuberculosis was not made, further studies confirmed it. The authors point to the existence of non specific pleuritis without neoplastic infiltration in some pleurisies consecutive to lung tumors or from other organs and to its therapeutic consequences. They also inform having found tuberculosis in patients over 50 years of age (8 in 11 cases). In this series the youngest pleural cancer case was a 27 years old man. PMID- 17424677 TI - [Ballistocardiographic changes in Argentinian hemorrhagic viral disease]. AB - Among eleven convalescent patients of Argentine hemorragic fever an electro ballistocardiographic study was effected which led to the following conclusions: a) In 2 over 11 studied cases, electro-ballistocardiographic modifications of probable myocardial injury could be detected. b) The greatest percentage of the modifications was interpreted as normal variants of the electrocardiogram. PMID- 17424678 TI - [Bronchial reactivity to histamine]. PMID- 17424679 TI - [Pachysteatosis of the scalp]. PMID- 17424680 TI - [Hydrothermal treatment of arteriosclerosis]. PMID- 17424681 TI - [Hydatid compression of the cervical cord]. AB - The clinics, L.C.R., mylographycs and nerosurgicals studies of a patient with an intradural cervical cord compression syndrom, multivesicular hydatidic, with primitive localization in the cervical vertebrae, are presented with anatomopathological verifycation. PMID- 17424682 TI - [In search of diabetic microangiopathy in the rectal mucosa]. PMID- 17424683 TI - Morel-Lavallee lesion. AB - Morel-Lavallee lesions, or closed internal degloving injuries, are most common in the subcutaneous tissues of the proximal thigh and trochanter. Bilateral lesions are rare. These post-traumatic fluid collections can be isolated or associated with concomitant pelvic fractures. Morel-Lavallee lesions may present as a pseudotumor when not recognized in the early post-traumatic period. Magnetic resonance imaging is the modality of choice in the evaluation of Morel-Lavallee lesions. The identification of a capsule on MRI can have important management implications. PMID- 17424684 TI - Allograft irradiation. PMID- 17424685 TI - Temperance in surgical training and technological advances. PMID- 17424686 TI - Hip arthroscopy for the treatment of children with hip dysplasia: a preliminary report. AB - The use of arthroscopic techniques to diagnose and treat hip disorders in children provides the potential to limit the morbidity of current techniques. PMID- 17424687 TI - The wheelchair axillary view of the shoulder. AB - The wheelchair axillary view of the shoulder allows a good image to be obtained in the seated position, and it can be used in the nonambulatory or multiply injured patient. PMID- 17424688 TI - Vacuum-assisted wound closure in postoperative spinal wound infection. AB - Combined with antibiotic therapy, vacuum-assisted wound closure may help reduce the need for serial irrigation and debridement surgery, contributing to a decrease in overall hospital stay. PMID- 17424689 TI - Delirium dilemma. AB - With understanding of diagnostic criteria as well as predisposing and precipitating factors, clinicians can impact the morbidity and mortality of delirium. PMID- 17424690 TI - Enzymatic activities in limb muscles subjected to external fixation with ring hybrid frames. AB - Enzymatic activities, which originate in the muscle envelope of tibiae with an experimental segmental bone loss, provide additional evidence for the intimate bone-muscle interrelationships in new bone formation. PMID- 17424691 TI - Bone health concerns in active and athletic women and girls. PMID- 17424692 TI - Preparing an orthopedic practice to survive a natural disaster: a retrospective analysis of rebuilding after Hurricane Katrina. AB - In the aftermath of the biggest natural disaster to hit the United States, valuable lessons can be learned to prepare us for future disasters. Physicians and other care givers should become involved in every level of disaster management. Most emergency plans are focused on triaging patients; however, little attention is paid to the logistics in evacuating hospitals or maintaining operations without outside communications. The lack of coordination and the breakdown of traditional communication channels were the biggest hurdles to overcome on the road to recovery. PMID- 17424693 TI - Tapered cementless femoral stem: difficult to place in varus but performs well in those rare cases. AB - This retrospective review was conducted to determine the incidence and minimum 5 year follow-up of varus placement of a tapered, proximally plasma-sprayed, titanium femoral component. Twenty-six (2.4%) of 1080 components were placed in > or = 5 degrees of varus in primary cementless total hip arthroplasty at one institution. Harris hip scores improved an average of 50 points. One stem was revised for malposition at 2.5 years, yet was well-fixed by radiographic criteria. Survival with aseptic loosening as an end-point is 100% with an overall femoral component survival of 96% at an average 10-year follow-up. A tapered, titanium component is reliably placed into appropriate position and is forgiving as to varus implant position. PMID- 17424694 TI - Reliability of measuring long-standing lower extremity radiographs. AB - The mechanical axis of the lower extremity was measured using preoperative long standing radiographs in 56 patients undergoing total knee arthroplasty with the Stryker Navigation System (Stryker, Kalamazoo, Mich). Each radiograph was measured by four physicians, three times, in random order, and at delayed intervals. Measurements from each physician produced intraclass correlation coefficients of 0.9872, 0.9973, 0.9958, and 0.9839. An intraclass correlation coefficient of 0.9844 was noted between each of the raters. Pre- and postoperative axial alignments were determined with Stryker Navigation. Preoperatively, each rater overestimated deformity approximately 1 degrees (P>.05); however, postoperatively each rater underestimated the degree of deformity by approximately 1 degrees (P>.05). Measuring long-standing lower extremity radiographs provides significant inter- and intrarater reliability in determining lower extremity alignment. PMID- 17424695 TI - Mechanical performance of hybrid Ilizarov frames containing full-threaded Schanz screws. AB - Modification of circular Ilizarov frames is necessary to overcome the difficulties in pin positioning due to unfavorable anatomic site or local soft tissue conditions. Hybrid frame configurations consisting of half pins or full threaded schanz screws are widely used in clinical practice. This study compared the mechanical performance of hybrid frames and a standard system. One standard and five modified hybrid systems were tested under axial compression, four-point bending, and torsional forces. Systems modified with full-threaded schanz screws showed a higher stiffness than half pin modifications and exhibited a similar mechanical performance of a standard system. PMID- 17424696 TI - Femoral head banking: NUH tissue bank experience. AB - National University Hospital Tissue Bank protocol follows guidelines recommended by the American Association of Tissue Banks and the European Association of Tissue Banks using donor selection criteria: medical history, clinical examination, chart review and laboratory tests for acquired immunodeficiency syndrome (AIDS), hepatitis B, hepatitis C, syphilis, and specimen for culture/sensitivity tests. For living donors, repeat testing is performed for AIDS and hepatitis C approximately 180 days after procurement. Femoral heads are procured using the "sterile double jar technique" and stored at -80 degrees C. Our first study of 273 consecutive potential donors undergoing hemiarthroplasty from 1989 to 1994 showed that a high percentage (42.5%) was unsuitable for use. A second study involving 175 potential donors was conducted from 1995 to 2003 after hepatitis C screening was introduced. The bacterial contamination rates in both studies (3.5% and 5.7%) are low. The incidence of other diseases also are low: hepatitis B, 2.3% and syphilis, 1.8% in the first study and hepatitis B, 5.7%; hepatitis C, 0.6%; and syphilis, 5.1% in the second cohort. No cases of AIDS were reported in either study. By 2003, femoral heads were transplanted in 205 patients with a low complication rate of 2.9%. PMID- 17424697 TI - Stress fractures in elite cross-country athletes. AB - This retrospective and comparative survey investigates an unusual number of stress fractures seen within a Division I college cross-country team. An anonymous questionnaire-designed to observe factors known to increase stress fracture incidence-was distributed to members of the current and previous seasons' teams. Running surface, sleep hours, intake of calcium, and shoe type were among the factors investigated. Eleven lower extremity stress fractures were found in nine athletes. Athletes with stress fractures reported significantly fewer workouts per week on the new track. All other study parameters had no statistically significant effect on stress fractures in these athletes. PMID- 17424698 TI - Osteochondral grafting of a central talar dome lesion using a wedge osteotomy of the distal tibia. PMID- 17424699 TI - Bilateral congenital absence of the flexor pollicis longus. PMID- 17424700 TI - Vancomycin-impregnated calcium phosphate cement for methicillin-resistant Staphylococcus aureus femoral osteomyelitis. PMID- 17424701 TI - [Rheumatoid arthritis: current status of therapy]. AB - Rheumatoid Arthritis (RA) is a frequent chronic inflammatory disease characterized by distal, bilateral and symmetrical lesions, leading to joint distortions and articular destructions. RA can also cause severe extra-articular manifestations associated with a poor prognosis. Recent advances in the field of immunopathology of RA have oriented treatment targeting the pro-inflammatory cytokines like tumor necrosis factor-alpha (TNF alpha), interleukin (IL) and IL6. These biotherapies are considered as an important therapeutic progress in the treatment of RA acting at the level of cellular processes responsible for rheumatoid disease. These new therapies are active not only in controlling the disease inflammatory processes but also to stop the radiological course of RA. These new therapies are however efficient as long as prescribed, their interruption being rapidly followed by a flare-up of RA. Multiple adverse events attributed to anti-TNF-alpha have been described especially severe opportunistic infections and tuberculosis. B cells playing a critical role in sustaining the chronic inflammatory process in RA, targeted depleting B cells therapies have been developed in refractory forms of RA giving promising results. However, before any biotherapy prescription especially of anti-TNF-alpha, an initial screening should be achieved to exclude patients with history of untreated tuberculosis, solid cancers, malignant hemopathies or demyelinating disorders. It is also essential to assure a strict follow-up in patients under biotherapy to detect adverse events that can be sometimes severe. Thus, the ratio benefit/risk must be evaluated before any biotherapy prescription. PMID- 17424702 TI - [Bibliometric profile of Tunisians medicals publications indexed in Medline from 2000 to 2003 part 2: social relevance]. AB - OBJECTIVE: This work, which is part of the medical documentary wakefulness, tries to evaluate the social relevance of the Tunisian medical publications indexed in the Medline's database. METHODS: This descriptive study covered the whole of the articles indexed in "Medline" between 2000 and 2003. We retained the medical papers whose author's address was a Tunisian health structure of either medical care or education with the first and / or the last author was a Tunisian doctor. The social relevance of the publications was measured by the agreement level of their content, studies through the "Major Essential Key Words" with the death causes and the morbidity global charge components, made by the World Health Organisation in its "report about the health in the world" in 2003. A "Major Essential Key Word" is a word chosen among the word's list "MeSH Major Topic" proposed by the bibliographic notice of Medline's database to summarize the essential topic of the article. RESULTS: The 1248 articles eligible to this study, have been indexed in "Medline" by "MeSH Major Topic" words using 3471 indexation lines. Among the "Top 20" list of the "Major Essential Key Words" only 9 themes were concordant with the community health needs, including four in relation with the epidemiologic "pre transition" epidemiology (tuberculosis, pregnancy complications, lung diseases, anemia) and five in connection with the epidemiologic "post transition" epidemiology (diabetes mellitus, breast neoplasms, leukemia, cardiovascular diseases, multiple myeloma). CONCLUSION: What emerges from this study is that medical research in Tunisia isn't concordant enough with the country epidemiologic priorities and public health national programs . The social relevance should be the principal selection criterion in the public financing of the research projects. PMID- 17424703 TI - [Severe corrosive oesophagitis : are high doses of methyl prednisolone efficient to prevent oesophageal caustic stricture in children?]. AB - BACKGROUND: Corrosive oesophagitis stricture is the long term complication of severe corrosive oesophagitis. The aim of our study was to evaluate the effect of a high doses of steroids on incidence and quality of oesophageal stricture. PATIENTS AND METHODS: We reviewed the case histories of 28 children seen at children hospital from 31 December 1991 to 31 december 2001. These children has second and third degree oesophageal burns and they were treated by systemic Methylprednisolone (1000mg/1,73/m2 SC). RESULTS: the frequency of stricture was 12/26 (46%). Ten children required A mean of 6,7 (5,74) dilatation range (1 - 17). One patient required an oesocoloplasty. The frequency of stricture in the group treated early before the 24th hour and after the 24th hours was (9/21) (47,4%) versus (3/7) (42,9%). This difference was not significant (P = 1). The frequency of stricture in the group treated less than 21 days and more than 21 days was 6/17 (40%) versus 6/9 (66,7%). This difference was not significant (P = 0,400). High doses of methyl prednisolone seems to decrease the risk of oesophageal stricture. We found no difference between the children treated before the 24th hours and those treated after the 24 hours and those treated less than 21 days and those more than 21 days. PMID- 17424704 TI - [The results of laparoscopic treatment of the gallbladder lithiasis. About 500 cases]. AB - BACKGROUND: Laparoscopic cholecystectomy is at present the gold standard treatment of gallbladder lithiasis. AIM: Assesment of Laparoscopic cholecystectomy METHODS: Through a retrospective series of 500 laparoscopic cholecystectomies during a period going from January 1996 to March 2000, we tried to evaluate our experience by comparing our results to the literature data. RESULTS: There were 420 women and 80 men with a sex ratio of 0.19. Average age was 50 years. 16.2% of our patients were obese. 13 patients had a history of respiratory disease and 122 history of a cardiovascular pathology essentially arterial hypertension. All our patients benefited from at least one hepato biliary ultrasound examination before the intervention. Antibioprophylaxy was administrated in 93.8% of cases. In 23 cases (4.6%), a conversion was necessary for different causes. We noted 11 surgical complications (2.2%) among which 2 required a surgical resumption, and 11 medical complications dominated by broncho pulmonary infections. We had no death. PMID- 17424705 TI - [Importance of CT-scanning in the evaluation of calcaneal fractures]. AB - The aim of the study is to evaluate the use fullness of CT in the calcaneal fractures. METHODS: We reviewed retrospectively 20 cases of calcaneal fractures collected between 2000 and 2003. RESULTS: We describe the different CT-scan patterns in 20 calcaneal fractures, according to Sanders classification. CT-scan helps for understanding fractures displacements, for choosing surgical treatment and evaluating patient prognosis. PMID- 17424706 TI - [Contribution of liver biopsy in intensive care unit. (180/2004)]. AB - AIM: Our aim was to evaluate the indications and contribution of liver biopsy (LB) in intensive care (ICU) and to compare them to those of LB in gastroenterology. METHODS: We included retrospectively 37 successive LB achieved in ICU and 38 successive LB achieved in gastroenterology. All data were reviewed by three intensivists and three gastroenterologists to determine the contribution of the LB. RESULTS: The indications of LB were different in the two units. The most frequent indications were cirrhosis (36.8%), isolated biological hepatic disruptions (26.3%) and histological classification of viral hepatitis (18.4%) in gastroenterology and isolated biological hepatic disruptions (48.6%), hepatopathy during pregnancy (27%) and fever of unknown origin (10.8%) in intensive care unit. According to the six reviewers, LB was enough contributive in the two units, (78.4% in ICU and 71.1% in gastroenterology -p=0.46-). It allowed to eliminate, to confirm, or to change a diagnosis in more than 70% of cases and allowed to modify the course of therapy in 21.6% of cases in ICU and in 26.3% of cases in gastroenterology (p-0.6). CONCLUSION: The LB is a feasible technique in ICU and can be as contributive as it is in gastroenterology. PMID- 17424707 TI - High-dose therapy and autologous stem cell transplantation for Hodgkin's lymphoma in relapse or failure after initial chemotherapy : results of the Centre National de Greffe de Moelle Osseuse de Tunis. AB - AIM: In the present study we report the clinical outcome of 27 patients with refractory or relapsed Hodgkin's lymphoma (HL) undergoing autologous peripheral stem-cell transplantation (ASCT). METHODS: On transplant, 18 patients had sensitive disease (SD) and 9 resistant disease (RD). The median time between diagnosis and ASCT was 18 months (range, 7 to 96 months). The conditioning consisted of BEAM regimen. RESULTS: The 100-day mortality rate was 3%. Three months after transplant, 12 patients transplanted with SD were in complete remission (CR) and only one of the 9 patients transplanted with RD achieved CR. Overall survival and disease-free survival after 3 years were 68% and 60%, respectively. CONCLUSION: the present results confirm the efficacy and safety of the ASCT in refractory or relapsed HL patients with SD. Other strategies should be investigated for patients with RD. PMID- 17424708 TI - [Fixed drug eruption : about 13 cases]. AB - AIM: To discuss, through a retrospective study, the epidemiological and clinical aspects and the causative agents of fixed drug eruption. METHODS: Thirteen cases were collected retrospectively during 11 years. There were 10 females and 3 males with a mean age of 44 years. RESULTS: The lesions correspond to erythematous plaques which fade to leave slate-brawn macules in all cases. The most frequent localizations were limbs (12 cases), trunk (6 cases), face (3 cases) ans external genitals (3 cases). Sulfonamides were the most frequent responsible drugs in our series (7 cases). CONCLUSION: Fixed drug eruption is characterized by one or more erythematous plaques which recur in the same places after challenge. Sulfonamides are actually the most frequent causative drugs in the different series. PMID- 17424709 TI - [Utility of stress cardiography to identify myocardial ischemia]. AB - BACKGROUND: The dobutamine echocardiography takes more and more an important place in diagnosis, prognosis and therapeutics strategies of the coronary disease. AIM: We prospectively enrolled 130 consecutive patients followed for a coronary insufficiency METHODS: the aim of determinating the diagnostic value of the dobutamine echocardiography in the myocardial ischemia compared with the classic means as the exercise test and the myocardial scintigraphy. RESULTS: Our results are comparable to the data of the literature, the dobutamine echocardiography is more specific than the exercise test and the myocardial scintigraphy (96 %, 61% and 53 %), more sensitive than exercise test (75 % versus 47 %) but less sensitive than the myocardial scintigraphy (75 % versus 96 %). The stress echocardiography has the best diagnosis precision 87% against 72 % for the scintigraphy and 56 % for the exercise test. The dobutamine echocardiography is a very feasible, tolerated well exam and especially very reliable in term of detection of the coronary disease in the feminine population, with a sensibility at 66,7 %, a specificity at 100 % and a diagnostic precision at 92 %, also in patient with hypertension the stress echocardiography is much more specific and reliable than the exercise test (100 % vs 22 % and 93 % vs 54 %, respectively),and it's superior to the exercise test in the presence of electric signs of systolic excess load and to the myocardial scintigraphy in case of left ventricular hypertrophy. The dobutamine echocardiography can be considered as safety and reliable means of investigation of the coronary insufficiency . PMID- 17424710 TI - [Sweet's syndrome: a propos of 8 cases]. AB - BACKGROUND: Sweet's syndrome is a dermatosis classically determined by painful erythematous plaques or nodules associated with fever, neutrophilia and dense neutrophilic dermal infiltrate. In most cases, Sweet's syndrome may occur in the absence of other diseases. However, it can be associated with an inflammatory disease and essentially with malignant conditions in 20% of cases requiring a careful investigation. AIM: We report herein, the epidemiological, clinical, therapeutic and evolutive features of a hospital serie. METHODS: We retrospectively reviewed all the files of patients conforming with the diagnosis of SS, seen at the dermatology departement of Habib Thameur hospital during a 7 year period (from 1997 to 2003). All patients fulfilled at least two major criteria and two minor criteria of Su et Liu's diagnostic criteria modified by Von Den Driesch. RESULTS: 8 cases of .Sweet's syndrome were diagnosed. All patients were females. The mean age was 51.62 years. The lesions occurred on the upper limbs in 7/8 cases. SS was isolated in 6 cases and para-inflammatory in one case corresponding to an associated Sjogren's syndrome. Besides the standard therapy using oral corticosteroids (3 patients), non steroidal inflammatory agents has been efficient in 4 cases. CONCLUSION: In our serie, we report an exclusive female involvement and a lower frequency of associated diseases compared with the relevant literature. Association of SS and Sjogren's syndrome is exceptional. To our knowledge, only 6 cases have already been reported. PMID- 17424711 TI - [Henoch Schonlein purpura in children: clinical and evolutive study of 122 cases]. AB - AIM: Analyze the clinical and evolutive particularities of Henoch Schonlein purpura in children METHODS: We studied retrospectively 122 cases enrolled in the pediatrics department of Sousse during 10 years period (1992-2001). RESULTS: It is about 66 boys and 56 girls (sex - ratio= 1.18) aged 3 to 13 years (mean age: 7 years and half). The diagnosis has been established clinically on the presence of cutaneous syndrome with symmetrical declivitous region purpura in all patients with articular syndrome (91cases) and/or digestive syndrome (65cases). Complications were variable: digestive hemorrhage (19 cases), occlusive syndrome (2cases), renal involvement at variable severity (56 cases), scrotal and testicular complications (11 cases), cardiac complications (tamponade in a case). Henoch Schonlein purpura was associated with a primary antiphospholipid syndrome in a case, renal tuberculosis in a case and cholestatic hepatitis A in another case. All patient receeved symptomatic treatment (rest in bed + / - antalgic treatment). Digestive rest was prescribed for 20 patients presenting severe abdominal pains with corticosteroid during 2 at 4 weeks (1-2mg/kg/d) in eight cases. Corticosteroid-cyclophosphamid association was prescribed for 2 patients with severe renal involvement; one of them benefitted of extra-renal purification. One or several relapses of Henoch Schonlein purpura were noted in 13 patients. All sick evolved favorably same those presenting renal or cardiac involvement (middle receding of 5 years). PMID- 17424712 TI - [Acute otomastoiditis in children, about 44 cases]. AB - BACKGROUND: Acute mastoiditis with ostertis is secondary to bacteria involving mastoid cavities AIM: Describe the epidemiologic aspects MATERIAL AND METHODS: Between 1990 and 2003, we listed 44 files of children hospitalized for acute mastoiditis. The treatment was in any case medical associated to a surgical treatment in 30 cases. RESULTS: The age of our patients was consisted between 2 months and 14 years with an average is 2 years. The sex ratio of patient was 1,1. 29 patients didn't have any antecedents particular O.R.L, 8 patients were followed for otitis middle chronic and 7 had an cholesteatomateuse. The auricular old-fashioned swelling was the most frequent functional sign. A scanner has been achieved in 13 cases. The treatment was medico-surgical in 30 cases is 68,8% and an only antibiotherapie in 14 cases is 31,8%. PMID- 17424713 TI - [Salivary gland biopsy : experience of La Rabta Hospital's pathology departement]. AB - BACKGROUND: The minor salivary glands biopsy is a very common diagnostic procedure in oral medicine rather its efficiency has not been statistically proved. AIM: Assessment of Rabta pathologic department experience METHODS: 297 biopsies have been studied with special attention to the suspected diagnosis before biopsy and the final histologic result. RESULTS: The minor salivary gland biopsy confirmed the initial diagnosis in 78 cases. Although if the minor salivary gland biopsy is in most cases not contributively, it is a very simple procedure which gives the diagnosis of Gougerot-Sjogren disease, amylosis and sarcoidosis. PMID- 17424714 TI - Epidemiological and medical aspects of epilepsy in the elderly. PMID- 17424715 TI - Primary leiomyosarcoma of the vagina. Case report and literature review. AB - BACKGROUND: The vast majority of primary vaginal cancers are squamous cell carcinoma. The leiomyosarcoma of the vagina is extremely rare neoplasm. It comprising 2% to 3% of vaginal malignancies. Only 77 cases have been reported up to date in Anglo-Saxon literature . AIM: Report of a new case CASE: We report a case of a patient 43 years old, single and presented a tumor of 7 cm to the detriment of the posterior wall of the vagina. The treatment consisted of a radical colpohysterectomy followed by postoperative radiotherapy. After 40 months, she is still alive without local or distant recurrence. CONCLUSION: This is a rare observation. The surgery is the basic treatment, the poor prognosis of these tumors remains linked especially to the histological grade. PMID- 17424716 TI - [Last discovery of a congenital broncho-oeso-phageal fistula]. AB - BACKGROUND: Broncho-oesophageal fistula is rare and occurs mostly in acquired condition. AIM: Report of a new case. CASE: We report a case of a 58-years-old woman which has, since 38 years ago, a chronic cough, recurrent bronchopulmonary suppurations and hemoptysis. Clinical examination let to the diagnosis of a broncho-oesophageal fistula. Surgical treatment resulted in complete recovery. There was no evidence of any aetiology. Congenital origin of the fistula was deduced. PMID- 17424717 TI - [Primary mediastinal seminoma. Report of three cases]. AB - BACKGROUND: Primary mediastinal seminomas (PMS) are rare tumors that are morphologically similar to their testicular counterparts but may have different biologic behavior due to their particular anatomical location. AIM: New cases report of PMS CASES: Three new cases of primary mediastinal seminoma are presented. The patients were men aged of 16, 33 and 47 years. Their clinical symptoms included cough, dyspnea, chest pain and superior vena cava syndrome. None of the patients had a previous history of testicular neoplasm or tumor elsewhere. Mediastinoscopic biopsy was performed and histological examination results revealed a seminoma. Immunohistochemical showed membranous staining with placental alkaline phosphatase in the three cases. Chemotherapy has largely replaced surgical resection and radiotherapy as the initial treatment in patients with mediastinal seminoma. PMID- 17424718 TI - [Retroperitoneal ganglioneuroma. A case report]. AB - BACKGROUND: Ganlioneuroma is a rare, benign, neurogenic tumor originating from the neural sheath and frequently localized in the retroperitoneum. AIM: Report of a new case. CASE: We report a case of a 36-year-old woman presenting isolated right lumbar pain. Computed tomography revealed a heterogeneous, right, retroperitoneal mass, 13 cm in diameter. Complete resection of the tumor and right nephrectomy were performed. Histology confirmed the diagnosis of ganglioneuroma. Follow-up was 10 months with no recurrence. Through this case, we discuss diagnostic, therapeutic and prognostic aspects of this disease. PMID- 17424719 TI - [Isolated adenomatous polyposis of the appendix. Report of two cases]. AB - BACKGROUND: Adenomas are rarely diagnosed in the appendix and may be isolated or may coexist with other neoplasms in the gastrointestinal tract. This emphasizes the need for postoperative colonoscopy when a polyp of the appendix is found. Moreover, the polyps are considered to be premalignant lesions. AIM: Report two new cases of adenomas of the appendix. CASES: We report the cases of 23-year-old and 22-year-old men, for whom appendicectomy performed for acute appendicitis. In both cases, histologic studies revealed adenomas in moderate dysplasia ; colonoscopy did not reveal any polyps. PMID- 17424720 TI - [Doppler echocardiographic diagnosis of ventricular asynchrony. Present situation and future perspectives]. AB - Heart failure is among the most common chronic diseases in the modem era. It is the major cause of morbi--mortality and hospitalization in patients aged more than 60 years. Dilatation of left ventricle frequently induces intracardiac conduction delays resulting. in asynchronous left ventricular motion. Showing as left bundle branch block in the surface EKG. Diagnosis of asynchrony based only on electrocardiogram criteria (wide QRS complex and left bundle branch block(LBBB)) proves to be non satisfactory considering the absence of parallelism between conduction unrests and cardiac dyssynchrony: 35% of LBBB are not associated with interventricular asynchrony and 20% without intraventricular asynchrony. So there is a necessity to define new criteria of asynchrony linked more directly to cardiac mechanical activity then EKG. Doppler Echocardiography brings more informative parameters concerning inter and intra ventricular asynchrony. Doppler tissue imaging can provide accurate information on electromechanical coupling and also assess interventricular and intraventricular dyssynchrony. It seems to be very promising for the prediction of the most satisfactory response to cardiac resynchronization therapy. PMID- 17424721 TI - The OPTN/UNOS Renal Transplant Registry. AB - The 10-year graft survival rates for first renal transplants performed during 1990-1994 and 1995-1999 and reported to the OPTN/UNOS Renal Transplant Registry increased from 57-58% for living donor transplants, from 42-46% for deceased donors aged 60 or under, and from 22-28% for donors over age 60 comparing the 2 intervals. These modest increases were accompanied by a 2% decline in 10-year patient survival for recipients of living and younger deceased donor grafts and a 1% improvement in patient survival for recipients of older donor kidneys. The 5 year graft and patient survival rates for transplants performed between 2000 and 2004 were 80% and 90% for living donor, 69% and 90% for standard criteria deceased donor and 55% and 82% for expanded criteria donor transplants, respectively. There was no significant improvement when compared to the 1995-1999 period for any of these groups and patient survival had declined by 1% among recipients of living or standard criteria deceased donors. recipients of living or standard criteria deceased donor kidneys had a 6-7% higher 5-year survival rate and longer graft half-lives than recipients of HLA mismatched kidneys. The number of local HLA-DR matched transplants (excluding zero-HLA-ABDR mismatched grafts) has been declining since 1998 and was affected by the activity of the local donation service area (presumably reflecting the size of the waiting list). There was a modest increase in the percentage of broadly sensitized recipients transplanted during 2002-2004 from 8-10% of standard deceased donor The median age for recipients of primary standard criteria deceased donor transplants increased from 43 during the period 1990-1994 to 51 during 2000-2004 and may explain the lack of improvement in long-term graft survival rates. When patients aged 19-35 were analyzed separately during the 3 periods, there was a 3-4% increase in actuarial or projected 10-year graft survival for recipients of living or younger deceased donor kidneys during each interval (p < 0.001). Changes to the kidney allocation algorithm that affect the role of HLA matching have not had a striking impact on the number or percentage of zero HLA-ABDR mismatched SCD transplants, which account for 16-17% of SCD transplants each year. The number and percentage of HLA-matched ECD transplants declined from 113 (12%) in 2001 to 63 (4%) in 2004. The 56% 5-year graft survival rate for recipients of HLA-matched ECD kidneys was not significantly better than that for HLA-mismatched grafts, whereas HLA-matched and from 2-4% of living donor kidney recipients that was temporally associated with improved technologies for detecting anti-HLA antibodies. The presence of panel reactive antibodies had almost no effect on 5-year graft survival among retransplanted patients. The number of transplants between spouses leveled off in 2001 at about 700 transplants each year. The number of non-spouse unrelated living donor transplants has increased 10-fold over the past 10 years to 1,341 in 2004 and does not appear to be slowing. PMID- 17424722 TI - Liver transplantation in the United States. AB - Based on the data reported to the OPTN/UNOS Liver Transplant Registry between 1987-2005, we found: 1. The number of deceased-donor liver transplantations increased slowly each year, with most of the increase being in adult recipients. The number of LD transplants, on the other hand, decreased sharply after 2002, following 3 years of rapid increase from 1998-2001 in both pediatric and adult recipients. 2. The number of DD liver recipients with non-cholestatic liver diseases increased very quickly during the past 18 years. Malignant disease as a cause of end-stage liver disease increased after implementation of MELD in 2002. Among LD liver recipients, non-cholestatic disease increased sharply from 1998 2001, but decreased from 2002. Malignant diseases as a cause for LD transplants decreased after 2002. 3. Among pediatric recipients, LD transplants provided better 5-year graft survival rates than transplants from deceased donors; in contrast, LD transplants in adults had poorer graft survival rates than those from deceased donors. 4. The use of marginal donors, including older donors, HCV (+) donors, donation after cardiac death donors, and diabetic donors, increased in the past 18 years. HCV(+) livers transplanted into HCV(+) cirrhosis recipients had similar graft survival when compared with HCV(-) donor livers, whereas when they were transplanted into non-HCV cirrhosis patients, they had poorer graft survival (60% vs. 70% at 5 years, respectively). When livers from diabetic donors were transplanted into diabetic recipients, they had much poorer graft survival than transplants from non-diabetic donors (54% vs. 77% at 5 years, respectively). 5. Split and partial liver transplants had poorer 5-year graft survival rates (58% and 57%, respectively) than whole liver transplants (62%), but the difference was mainly due to poorer outcomes during the first posttransplant year. 6. PELD allocation has resulted in improved one- and 3-year graft survival rates among pediatric liver recipients. Among adults MELD-based allocation has resulted in better one-year survival rates. When comparing the different original diseases, only HCC patients showed better one- and 3-year graft survival rates after MELD. 7. Within one year after transplantation, primary non-function and infection were the major causes of graft failure. These decreased after 1996, but recurrent hepatitis has increased as a cause of graft failure. After one year, chronic rejection and infection had decreased, while hepatitis recurrence still increased. 8. Cardiovascular deaths and deaths from multiorgan failure that occurred within the first year after transplantation have increased since 1996, while deaths due to infections have decreased. After the first year, deaths from graft failure increased, while CVD and infections decreased. PMID- 17424723 TI - Lung transplantation in the United States: 1990-2005. AB - The number of lung transplants reported to the OPTN/UNOS Registry has been increasing during the past 15 years. The increase is mainly due to a steady increase in transplants in the 50-64 age group. There is also a trend toward increasing lung transplants for older patients (> 65 yr) since 2001. Among whites, COPD, IPF, CF, alpha1-ATD, and PPH were the top 5 original diseases leading to lung transplantation; while for blacks, sarcoidosis, COPD, and IPF were the major original diseases. Recipients with IPF had significantly lower graft survival rates than COPD patients. One-year graft survival rates increased from 70% in the early 1990s to more than 80% in the most recent 3-year period, but the increase in long-term graft survival has been very limited. The negative effect of previous transplantation on graft survival was only significant within the first post transplant year; in contrast, the negative effect of HLA mismatches is more apparent on long-term graft survival. Acute rejection within the first year had a significant deleterious effect on long-term graft survival. PMID- 17424724 TI - Development and current status of ECD kidney transplantation. AB - The worsening shortage of donor kidneys for transplant and the aging of both the donor and candidate populations have contributed to the increasing importance of ECD kidney transplantation. While ECD transplants have an increased risk of graft failure, for most candidates patient survival is still improved over remaining on dialysis. Because of this risk, however, ECD kidneys have a high likelihood of discard; significant geographic variation in discard and transplant rates impedes maximum utilization of these kidneys. The ECD allocation system was implemented to help facilitate expeditious placement of ECD kidneys to pre-consented candidates by a simplified allocation algorithm. Under this system, recovery and transplantation of ECD kidneys have increased at rates not seen with non-ECD kidneys and not predicted by preexisting trends. More disappointing has been the lack of effect on the percentage of discards and DGF, despite significant reductions in CIT. The disadvantage in graft survival for ECD kidneys extends equally across the spectrum of recipient characteristics, such that no one group of candidates has a proportionately smaller increase in risk. However, benefit analyses comparing the risk of accepting an ECD kidney versus waiting for a non ECD kidney demonstrate a significant ECD benefit for older and diabetic candidates in regions with prolonged waiting times. The potential value of an ECD kidney to an individual candidate hinges upon the ability to receive it substantially earlier than a non-ECD kidney. Thus, future allocation efforts may focus on ensuring that is the case. In allocation driven by net benefit, ECD kidneys may become an alternative for those who might not otherwise receive a kidney transplant. PMID- 17424725 TI - Application of diagnostic classification algorithms to identify the most common causes of post-transplant complications. AB - ICD-9-CM diagnoses for hospitalizations occurring during the first 6 post transplant months were summarized into Clinical Classifications System (CCS) categories. Of the 28,900 patients examined, 54% had at least one hospitalization. There were 2.39 hospitalizations per patient-year at risk. The total Medicare inpatient costs were $339 million and mean length of stay was 8.3 days. The most common and costly CCS diagnosis was complications of a kidney transplant followed by infections, circulatory system disease, gastrointestinal disease and endocrine complications. The CCS is a useful tool to summarize the complexity of claims information in the USRDS and could prove useful in further claims research. PMID- 17424726 TI - Report from the National Transplantation Pregnancy Registry (NTPR): outcomes of pregnancy after transplantation. AB - Experience in the field of pregnancy after transplantation has been gained through continued case reports, center reports, and registry data. The NTPR maintains an ongoing active database to study the safety of pregnancy and includes the outcomes of female transplant recipients as well as male recipients who father pregnancies. Analyses are ongoing and include long-term follow-up of recipients' graft status and of their offspring. For the most part, guidelines proposed in 1976 for counseling recipients remain applicable. While these counseling guidelines were formulated for kidney recipients, they may be extrapolated for other organ recipients as well. Organ-specific issues should also be considered in managing and counseling female transplant recipients. Recipients should be in general good health and graft function should be stable and ideally rejection free. They should have optimal control of comorbid conditions such as hypertension and diabetes prior to conception. While the shortest safe interval from transplant to conception has not been established, one year is a reasonable milestone, given the prerequisites of stable, adequate graft function and maintenance level immunosupression. During pregnancy, stable medication regimens should be changed as little as possible, and close maternal and fetal surveillance are required. These pregnancies are high-risk and require coordinated care among maternal fetal medicine specialists and transplant personnel. The pregnancy issues that face recipients and caretakers with the current adjunctive therapies and differing combinations of immunosuppressive regimens continue to require further study. The most pressing issue is the question of whether fetal exposure to mycophenolate mofetil or sirolimus confers additional risk, relative to the potential improvement in maternal survival and maternal graft function/survival conferred by these drugs. Given the multiplicity of immunosuppressive regimens, only broad-based registry participation can provide the data needed to analyze such complex questions. Future analyses are directed at potential effects of these newer immunosuppressive regimens, not only from immediate exposure, but also from potential long-term exposures such as may occur from breastfeeding. As the registry study design allows for continuing contact between registry staff and recipients and their health care providers, efforts are in progress to analyze long-term outcomes of parent and child. Continued close collaboration among specialists will help to better identify potential pregnancy risks in these populations, especially as new immunosuppressive agents are developed. PMID- 17424727 TI - The Bergamo Kidney Transplant Program. AB - Since the beginning of transplant activities in 1989, the Kidney Transplant Center at the Ospedali Riuniti Bergamo has based its clinical program on the most recent achievements of transplant medicine, in order to optimize the outcome of kidney grafts and improve the quality of life of kidney transplant recipients. Although the transplantation community attempts to keep up with increasing demand for transplantable organs, the supply continues to fall far short of the need. This observation prompted us to focus on the expansion of the available pool of deceased donor organs. In 1997, we established a dual kidney transplant program for donors older than 60 years based on a pretransplant histology protocol with a scoring system ranging from low-dose RATG and delayed CsA administration has been successfully adopted in this population of kidney transplant recipients in our routine clinic practice. In kidney transplantation, chronic deterioration of renal function and death with a functioning graft, mainly due to side effects of the medications, represents a major limitation for long-term success of many transplant programs. We recently documented that per-protocol biopsy more than one year after kidney transplantation is a safe procedure to guide change of conventional immunosuppressive regimens and to lower the risk of major drug related side effects. In particular, substantial reduction of the CsA dose, leading to extremely low CsA trough level, has no major detrimental effect on renal function and histology during 3 years follow-up, while patients remain free of rejection episodes with concomitant steroid and azathioprine therapy. Novel induction therapies with Campath-1H or Simulect and low-RATG have also helped to minimize maintenance immunosuppression in most patients largely avoiding the use of corticosteroids, Monitoring a patient's exposure to immunosuppressive agents is a critical issue in a minimum of 0 (no renal lesions) to a maximum of 12 (marked changes in renal parenchyma). The assumptions of the proposed algorithm to guide acceptance of single suboptimal or dual marginal kidneys for transplantation were validated in a prospective pilot study involving centers in Europe and North America. Whether the encouraging short-term data translate into improved graft survival is currently a matter of investigation in a prospective, multicenter, matched-cohort trial. As kidneys from marginal donors have an increased risk of delayed graft function, we also studied strategies to manage and prevent this complication. A dual immunosuppressive regimen of basiliximab and transplantation. By pharmacokinetics studies, we documented that a fixed dose regimen of MMF--adopted in the majority of transplant units worldwide--might no longer be the best approach for the management of transplant patients, and MPA pharmacokinetic monitoring is advised. Similarly, we reported pharmacokinetic interaction of concomitant immunosuppression on blood levels of the new immunosuppressant sirolimus. We have a special multiorgan transplant program at our center for patients affected by rare diseases, such as the recurrent hemolytic uremic syndrome (HUS). Based on genotyping for complement factor H-1, membrane co-factor protein or factor I gene mutations, we are exploring the possibility of combining liver and renal transplant or performing renal transplant alone in patients with recurrent HUS who have end-stage renal disease. The achievements of our clinical center are the result of the continuous support by an intense clinical and basic research program. This has allowed us to create a unique model to address the major challenges of transplant medicine. PMID- 17424728 TI - Long-term outcomes of kidney transplantation in recipients 60 years of age and older at the University of Florida. AB - As the population ages, the transplant community will continue to see "elderly" patients with end-stage kidney disease who are seeking transplantation. In this report we describe long-term outcomes of 315 primary kidney transplants performed at the University of Florida in recipients aged > or = 60 years and compare them to results from 3 younger recipient cohorts. Among recipients > or = 60 years, patient survival was significantly worse than for younger recipients but no differences in graft or death-censored graft survival were seen. We suspect that although patient survival was worst in the oldest group, there were likely other causes of graft loss within the younger groups that balanced the effects of death on graft survival in the oldest group. Among recipients aged > or = 60 years, patient survival at 10 years was 55% for living-donor kidney recipients and 46% for deceased-donor kidney recipients. African-American recipients had a higher risk of mortality and graft loss in all age groups after deceased donor kidney transplantation but not after living donor transplantation. Delayed graft function negatively impacted outcomes among all recipients and the adverse effects were greater after deceased donor than living donor transplantation. These effects were also seen within the oldest recipient age group. Increased donor age was a significant risk factor for death and graft loss among all age groups after deceased donor kidney transplantation but not among living-donor kidney recipients. More specifically, recipients aged > or = 60 years who received kidneys from donors > or = 60 years demonstrated significantly worse outcomes when compared to those receiving donor kidneys < 60 years. The presence of diabetes mellitus in recipients > or = 60 years was not a significant risk factor for mortality or graft loss after transplantation. Acceptable results can be obtained after kidney transplantation in recipients aged > or = 60 years. Future investigations should focus on improving recipient selection in the elderly population, identifying strategies to minimize DGF in deceased donor kidneys, understanding all variables involved in the risk associated with recipient race, and increasing living donor transplantation across all age groups. PMID- 17424729 TI - Evolution of immunosuppression in kidney transplantation: the Ohio State University experience. AB - The goals and outcomes of immunosuppression in renal transplantation have changed significantly over the last 30 years. When graft survival rates were relatively low and acute rejection was a frequent occurrence in the early era of transplantation, the goal of immunosuppression was to improve survival and reduce the rate of acute rejection. Today, with excellent graft survival rates and a low incidence of acute rejection, the goal of immunosuppression has shifted toward not only eliminating acute rejection, but also toward reducing the side effects of medications, and maintaining long-term graft function by decreasing chronic nephropathy. Between September 1982-December 2004, 3,211 primary kidney transplant procedures were performed at The Ohio State University. We excluded from analysis all combined transplants as well as patients who were involved in clinical research protocols. Our immunosuppressive protocol changed substantially over this 24-year period, which can be divided into 5 eras in time. Each era is defined by a distinct immunosuppressive protocol that resulted in an incremental improvement in outcomes of patient and graft survival rates. In the present study, the outcomes of each era in patients with previous kidney transplant only are compared and future directions are discussed. The incidence of acute rejection episodes and graft survival from each era are compared and demonstrate the substantial improvement in results that have been achieved over the past 24 years. PMID- 17424730 TI - The Kidney Transplant Program at King Faisal Specialist Hospital and Research Center. AB - More than 1,000 kidney transplants were performed at King Faisal Specialist Hospital and Research Center (KFSH&RC) between 1981-2005. The majority were from living donors. The renal transplant program at KFSH&RC was fundamentally transformed in 2001 with the introduction of renal transplant physicians and the emphasis on multidisciplinary teamwork. This fundamental change has resulted in tripling of the size of the program and in expanding its scope of services to include high-risk patients (highly sensitized with a positive crossmatch). These achievements were coupled with excellent outcome data. The 5-year patient and graft survival rates for adult transplants performed during 2000-2005 were 97% and 94%, respectively, for 268 living donor transplants and 97% and 76%, respectively, for 73 deceased donor transplants. The kidney transplant program at KFSH&RC is now a leading center in the Region and it ranks among the leading kidney transplant centers in the world in terms of size, scope of services and outcomes. PMID- 17424731 TI - Solid organ transplantation at the National Institutes of Health: development of a research-based transplantation practice. AB - The National Institutes of Health has established a clinical transplant research program focusing on translational research in kidney transplantation. The program has been developed with a multidisciplinary approach under a common administrative structure that integrates transplant physicians and surgeons with clinical laboratory and data analysis support personnel. The program has achieved excellent clinical outcomes despite focusing exclusively on investigational methods and serving a diverse and medically complex patient population. Novel approaches toward consenting, computer integration, and tissue acquisition have been layered over interventional and observational studies to serve the scientific mission while delivering quality transplant care. PMID- 17424732 TI - Organ transplantation at the Medical University of Warsaw. AB - The Warsaw Transplant Center comprises 3 programs for transplantation of kidney, pancreas and liver. At the end of 2005, 3,616 kidney, 131 simultaneous pancreas kidney and 592 liver transplants had been performed. The one-year patient and graft survival rates were 92.8% and 91.1%, respectively, for 2,689 kidney transplants performed in 2001-2003 and the 5-year patient and graft survival rates were 91.4% and 77.1%, respectively, for 1,667 transplants performed in 1998 2001. The number of liver transplantations performed at the Medical University of Warsaw is growing. The gender distribution is well balanced and the most common age of the transplant recipients ranges from 41-50 years. The most common indications were post-hepatitis C and B cirrhosis and post-alcoholic cirrhosis. The average one-year mortality after liver transplantation was 9.8%. The results of liver transplantations at the Medical University of Warsaw are similar to those reported by other leading European centers. At present, 495 patients are alive with good liver function 1-8 years after transplantation. PMID- 17424733 TI - Clinical islet transplantation at the University of Alberta--the Edmonton experience. AB - While the field of islet transplantation has evolved over the past 30 years and exponential progress and increase in clinical activity has occurred during the past 5 years, it is clear that major challenges still remain, particularly in understanding why islet function seems to decay over time. High one-year rates of insulin independence, and high 5-year rates of partial islet function (with C peptide secretion and protection from hypoglycemia) are now routine. Improved control of glycated HbA1c and reduced risk of recurrent hypoglycemia are benefits of islet transplantation irrespective of the status of insulin independence. If complete and sustained freedom from insulin is the primary objective, it is clear that whole pancreas transplantation still offers far superior metabolic reserve. However, the less interventional nature of islet infusion and avoidance of major surgery are advantages of islet transplantation over whole pancreas strategies. While the anti-rejection drugs available today may have had an acceptable safety profile in most islet transplant recipients, the drug-related and dose-limiting side effects have proved to be a challenge in some patients. Current islet-alone transplantation requires lifelong immunosuppression and is limited to patients with recurrent severe hypoglycemia and severe labile diabetes. More effective treatments are needed to control both acute rejection and recurrent autoimmunity. Remarkable opportunities lie ahead for improved islet survival, better engraftment and the possibility of expansion of islet mass both in culture and possibly within the patient after transplantation. Living-donor islet transplantation offers one option to expand the available donor supply, but remains controversial because of the potential for diabetes induction or other morbidities in a healthy donor. The development of less toxic immunosuppression and perhaps immunological tolerance will one day also have a huge impact on this field. Alternative tissue sources from either xenogenic sources or stem cells will ultimately solve the challenge of limited donor supply. PMID- 17424734 TI - The University of California at Los Angeles heart transplantation experience. AB - During the past 20 years, the number of older patients undergoing heart transplantation has steadily increased as a result of expanding indications for this procedure. At UCLA, 1,046 adult heart transplant procedures were performed from 1984-2004. Actuarial one-, 3-, 5-year survival rates for all recipients are 83%, 76%, and 72%, respectively. Our patients were grouped into 2 eras: those transplanted between 1984-1993 and those between 1994-2004. The current era, incorporating those patients transplanted between 1994-2004, may be defined by the introduction of pravastatin in 1994, which we have reported to benefit heart transplant patients. There were 403 adult heart transplant recipients in the pre 1994 era and 643 adult recipients in the current era. Additionally, patients were then grouped by age into those aged 18-61 years and those older than age 61 years. In the current era, there was less rejection and cardiac allograft vasculopathy (CAV) with improved survival and comparable survival in younger versus older patients. Although cellular rejection has decreased over the 1990's decade, we have reported that the incidence of noncellular or humoral rejection in our cardiac transplant recipients has remained unchanged despite improved immunosuppressive therapies. Thus, there appears to be a need for newer immunosuppressive agents to treat humoral rejection effectively. CAV is one of the major factors limiting long-term survival in heart transplant patients. Early CAV can be detected by intravascular ultrasound (IVUS), which is a new technology that detects intimal thickening in the donor coronary arteries. We demonstrated in the multicenter IVUS validation study that the progression of IVUS-defined intimal thickening > or = 0.5 mm in the first year after heart transplantation appears to be a reliable surrogate marker for subsequent mortality, nonfatal major adverse cardiac events, and the development of angiographic CAV through 5 years follow-up. The limitation on the number of transplants performed has been the number of donor organs available. We began the alternate list at our program in 1992, which is designed to match those patients excluded for regular heart transplant listing (mostly older patients) to marginal donor hearts which are unused. We have demonstrated that the alternate list patients who have undergone heart transplant have satisfactory outcomes. This has allowed expansion of the donor pool and offered heart transplantation to those patients who would not routinely have an opportunity for this life extending procedure. PMID- 17424735 TI - Kidney transplantation at the Chinese People's Liberation Army General Hospital. AB - Between October 1977 and December 2004, 2,037 kidney transplants were performed in 1,804 patients with ESRD at the Peoples Liberation Army General Hospital Postgraduate Medical School. Overall graft survival rates at one, 5, 10, 15, and 20 years were 91.6%, 79.3%, 64.3%, 53.8%, and 47.6%, respectively. The number of transplants significantly increased during the past decade. One-year patient and graft survival rates have increased from 40% and 35% in the 1970s to 98% and 96% in the 2000s, respectively. Since CSA was introduced to our clinic, one-year graft survival rates have increased from 33% to 93% and the half-lives have significantly improved from 13.1 years to 21.7 years. Also, the graft survival rate for patients on CSA-based triple therapy using MMF was about 18% higher at 5 years than those for patients using AZA (92% vs. 74%, p < 0.001), and 11% higher at 10 years (71% vs. 60%, p < 0.01). Our analysis showed that transplant year, DGF, rejection, immunosuppressive regimen, ABO blood group, and original disease were independent factors impacting on graft survival and poor HLA matching with 5 6 mismatches had an adverse effect on graft survival compared with 1-2 mismatches. Infection, cardio- and cerebral accidents and hepatic failure were the 3 main causes of death in transplant recipients. PMID- 17424736 TI - Kidney transplantation at the Beijing Friendship Hospital from 1972-2005. PMID- 17424737 TI - Thirty years of kidney transplantation in two Chinese centers. AB - Since the first kidney transplantation was performed in 1976, 4,306 renal transplantations have been performed at Shanghai No. 1 People's Hospital and Fujian Oriental Hospital. Nearly all (99%) of the grafts were from deceased donors. Graft survival rates in the era of modern immunosuppression and HLA-based organ allocation have improved substantially. The one-, 5- and 10-year graft survival rates for 2,575 transplants performed during 1995-2004 were 93.9%, 71.8% and 53.9%, respectively. Chronic rejection accounted for 54.3% of graft failures and 54.2% of patient deaths resulted from graft failure. We performed 48 combined liver-kidney transplants since 2001 and the first simultaneous human islet-kidney transplantation last year. PMID- 17424738 TI - Twenty-nine years experience of kidney transplantation from Zhejiang University. AB - During the last 29 years the dramatic changes that have taken place in our transplant programs and in solid organ transplantation permit us to face tomorrow with courage and confidence. In the future we will be deeply concerned that the long-term benefit of more effective immunosuppression or novel strategies might be unfavorably impacted by the tendency to use organs whose intrinsic lifespan may be limited. We are deeply concerned by the inexorably increasing discrepancy between supply and demand for deceased kidneys, especially for the added morbidity that this inevitably entails for those patients with ESRD who do not have live donors. Our program continues to seek better ways to improve survival and the quality of the life of our patient population. PMID- 17424739 TI - The Organ Transplant Institute of the China Medical University. AB - The clinical transplant program at the First Clinical Hospital in CMU includes renal, hepatic, pancreatic-renal and hepatic-renal transplantation and is supervised by experienced transplant surgeons, transplant physicians and one laboratory team. Our center has formed a systematic management model for multi organ transplantation. We maintain a laboratory for HLA testing and immunological monitoring in accordance with international standards, have a computerized network database to collect and store patient information, provide individualized treatment, direct postoperative care and healing and insist upon long-term patient tracking and clinical follow-up. By the end of 2005, we had performed 604 kidney transplantations, 122 liver transplantations, 22 simultaneous pancreas kidney transplantations (SPK) and 4 combined liver and kidney transplantations. The one- and 5-year kidney graft survival rates were 96% and 84%, respectively. PMID- 17424740 TI - Liver transplantation at Tianjin First Central Hospital. AB - Liver transplantation has expanded rapidly in China. The Tianjin First Central hospital is currently the largest liver transplant center in China. During 2005, this center performed 647 liver transplants and 436 kidney transplants. Since the liver transplantation program began in 1998, this center has performed 1,803 liver transplants and 1,233 kidney transplants. Significant progress has been made during the past 5 years in surgical technique, immunosuppression and patient care after transplantation. Graft survival rates have also improved significantly. The one-year graft survival rate for 843 patients who received a liver transplant between 2001 and 2004 was 87.3%. About half of these patients suffered from hepatocellular carcinoma. We will continue to evaluate donor and recipient factors to maximize outcomes of liver and kidney transplantation. PMID- 17424741 TI - Blood and marrow transplantation in the People's Republic of China. PMID- 17424742 TI - Donation after cardiac death and the science of organ donation. AB - The science of organ donation is now well established for comparative analyses that will foster improvement in practices throughout the United States and these metrics could be used as a model for an international assessment of organ donation as well. PMID- 17424743 TI - Trends in donation after cardiac death. AB - The deceased cardiac donor (DCD) can provide organs that are suitable for transplantation. However, in order to increase recovery and utilization of this under-utilized organ donor source, a number of issues remain to be resolved. First, the public understanding is essential for support of this end-of-life option to donate after cardiac death. Extensive education of donor hospitals will be needed to identify potential donors and arrange for their care. Organ procurement organizations will have to commit to the extra effort required to identify and manage these donors and to design recovery techniques to maximize procurement of abdominal and thoracic organs. Pulsatile machine perfusion may be needed for assessing viability of DCD kidneys, although the evidence is lacking that pump preservation improves survival of DCD kidneys that are transplanted. It will also be important to identify transplant centers and suitable patients within those centers that will accept DCD organs. Preselecting recipients will speed allocation as has been the case for expanded criteria donors. Special efforts to minimize reperfusion injury should be made initially to protect what might be more "injury-prone" DCD organs and calcineurin inhibitors should probably be avoided during the early posttransplant period. The graft survival rates for DCD kidneys and pancreata have been comparable to those for conventional deceased donor grafts, whereas graft survival rates for DCD liver recipients have been significantly poorer than with conventional livers. Thus, we need to continue exploring approaches to improve patient and graft survival for DCD liver transplant recipients. PMID- 17424744 TI - Establishment of a nationalized, multiregional Paired Donation Network. AB - Over the past few years, significant progress has been made in the science and development of paired donation. With increasing awareness of paired donation and ready availability of the tools necessary to establish new consortia, paired donation can be made available to transplant programs and patients with increasing alacrity. Increasing registration of recipients and their donors for paired donation will lead to larger pools for matching and to transplantation of increasing numbers of patients via paired donation. As paired donation becomes common practice throughout the US and the international transplant community, its role in facilitating transplantation of sensitized patients will be better defined. Presently, paired donation remains an attractive alternative to desensitization and wait list paired donation for a majority of patients with preexisting humoral immunity to their donors. PMID- 17424745 TI - The changing role of antibody testing in transplantation. AB - Although it was established in the 1980's that positive crossmatches associated with adverse transplant outcomes were those due to HLA specific antibodies, the cell-based assays for determining antibody specificity were time consuming, laborious, and moderately specific and sensitive. Technological advances now permit much more rapid and thorough antibody characterization. While no single assay can identify all anti-HLA antibodies in a patient's serum, the repertoire of specificities that can be defined is impressive and continues to improve. By using complimentary assays, most predominant specificities can be defined in even very highly sensitized patients. Perhaps, more critical, the sensitivity of solid phase immunoassays, which in some cases surpasses that of cell-based flow cytometry, provides a new level of assurance in the detection of humoral sensitization. The impact of these changes in antibody testing is already being felt in transplantation, providing a better evaluation of the level and breadth of sensitization prior to transplantation and greatly facilitating the success of humoral desensitization protocols. Thorough antibody identification can also provide a "virtual crossmatch" or a profile of incompatible donors defined by unacceptable antigens. An approach using the frequencies of unacceptable antigens to derive the probability of incompatible donors will likely be a feature of revised renal allocation for sensitized patients in the US. A compelling body of data is also accumulating proving the efficacy of monitoring for the recurrence or de-novo production of anti-donor HLA specific antibodies after transplantation. The possibility of early detection of humoral rejection in time for effective clinical intervention may offer a means to combat the inexorable loss of grafts to chronic rejection. Looking further into the future, as extensive antibody definition and monitoring are more universally applied, the ability to rule out HLA specific antibodies as a cause of graft loss will certainly also help determine the role of non-HLA antibodies in the outcome of solid organ and hematopoietic stem cell transplantation. PMID- 17424746 TI - The needs for a Global Alliance for Transplantation. PMID- 17424747 TI - The Gift". PMID- 17424748 TI - Annual literature review for clinical transplants 2005. PMID- 17424749 TI - Revisiting the center effect. AB - Based on a model analyzing multiple effects, center variations in one-year outcomes from the OPTN/ UNOS Transplant Registry were used to estimate scores for 246 centers reporting 40+ renal transplants during 1995-2005. The adjusted one year graft survival (derived from scores and 5 fundamental covariates) varied from 72% to 99%, demonstrating that the center effect is still the most influential factor in early renal graft survival. The 5 fundamental variables (recipient/ recipient's pre-transplant waiting time, original disease, age and degree of sensitization (PRA), proportions of expanded versus standard criteria donor grafts, donor age, cold ischemia time, transplant CMV and flow cytometry crossmatch statuses, and type of induction therapy. The associations indicated that centers with higher scores tended to transplant "better" kidneys into donor sex and race and transplant year) combined to reduce center variation by -10%. After categorizing centers into 5 groups based on increasing scores, we analyzed the extent to which groups differed over 15 secondary factors related to primary, deceased-donor transplant survival. All variables were statistically associated with center groups, but not all of the significant differences were clinically interesting. The notable associations included "more favorable" recipients. However, one-year graft survival rates still increased stepwise with increasing center success at nearly all levels of secondary variables. Thus, differences in center-specific success rates were not the direct result of center practices with regard to known kidney transplant factors. PMID- 17424750 TI - Alemtuzumab induction in kidney transplantation. AB - The use of alemtuzumab for induction therapy is increasing. 7.6% of antibody induction recipients in 2003 and 9.3% of induction recipients in 2004 received alemtuzumab. Antibody induction, especially T-cell depleting agents, Thymoglobulin and alemtuzumab are very effective in preventing early acute rejection in the initial hospitalization for both deceased and living donor transplants. However, when examining the effect of antibody induction on acute rejection after the initial hospitalization during the first year, alemtuzumab induction was associated with increased odds of acute rejection in deceased donor transplants compared to no antibody induction, IL-2RA, and Thymoglobulin. There was no difference in acute rejection during the first year after initial hospitalization in living donor transplants among all 4 induction groups. Despite lower acute rejection rates with alemtuzumab induction, no difference in graft survival was observed compared to no antibody induction, IL-2RA, and Thymoglobulin over 24 months. We conclude that alemtuzumab is an effective induction agent in kidney transplantation. However, further studies are needed to assess its long-term efficacy and to establish the optimal immunosuppressive regimen that should be maintained when alemtuzumab is used as an induction agent. PMID- 17424751 TI - Trends in serum creatinine levels and transplant year. AB - The serum creatinine (SCr) level has a strong predictive value for kidney graft survival. On average, the SCr levels of recipients whose grafts continued to function were lower (< 1.5 mg/dl) than those for recipients who died with a functioning graft during the following year (-2.0 mg/dl) and were much lower than those for recipients whose graft failed within a year due to chronic rejection ( 3.0 mg/dl). The average one-year SCr levels have fallen steadily since 1995 among recipients of living and deceased donor kidney transplants, suggesting that immunosuppression and clinical management have improved graft function. These improvements have come despite increasing transplantation of older donor kidneys and heavier recipients, both of which are factors predisposing to higher SCr levels. Even though the average one-year SCr levels have dropped over the past 10 years, there has not been a substantial improvement in long-term graft survival. PMID- 17424752 TI - Immediate extubation in the operating room after cardiac operations with thoracotomy and sternotomy. AB - BACKGROUND: Immediate extubation after cardiac operations in the operating room (OR) is rarely practised in Taiwan. The increased use of the minimally invasive and off-pump coronary artery bypass surgery (CABG) and the financial pressure from health insurance have raised the interest of its application after cardiac operations. The purpose of the study was to investigate the practice of immediate extubation in patients undergoing cardiac operations via minimal invasive thoracotomy against via midline sternotomy. METHODS: Seventy unselected consecutive patients undergoing cardiac operations via either minimally invasive thoracotomy or midline sternotomy were enrolled for investigation. Anesthetic management, including bispectral index and continuous train-of-four (TOF) monitoring, was modified to extubate the patients in the OR within 15 min after the closure of the skin wound. Extubation criteria based on clear consciousness, recovery of muscle power, and stable hemodynamics without purposeful strong inotropic support were stipulated. Patient-controlled analgesia was used to provide adequate postoperative pain control. RESULTS: Extubation within 15 min after the end of surgery was successful in all patients who underwent thoracotomy while there were five patients who could not be extubated in the sternotomy group. The PaCO2 values 30 min after extubation and the ICU stay were significantly higher and longer in patients of the sternotomy group. The pain intensity after extubation or the doses of analgesics used did not differ between two groups of patients. There was only one patient in the sternotomy group who needed reintubation and there was no postoperative death in both groups of patients. CONCLUSIONS: We consider that immediate extubation after cardiac procedures either through thoracotomy or sternotomy is a rather safe practice and if possible minimally invasive technique should be chosen because it causes smaller wound and offers safer immediate extubation and shorter ICU stay. PMID- 17424753 TI - Comparisons of the dose-response and recovery time course of vecuronium and atracurium in anesthetized chinese adult patients. AB - BACKGROUND: The authors found no study comparing potency and duration of action of vecuronium and atracurium in healthy Chinese, so they compared the dose response relations of vecuronium and atracurium in anesthetized Chinese adult patients and evaluated the time-course of action of two drugs following administration of equipotent doses. METHODS: Sixty patients, ASA grade 1, aged from 17 to 50 yr, scheduled for elective plastic surgery were randomly distributed to either the vecuronium or the atracurium group. General anesthesia was maintained with 60% nitrous oxide in oxygen (N2O-O2), thiopental and fentanyl. Neuromuscular function was assessed using a accelerometer with train-of four (TOF) stimulation at the wrist every 12 sec, and the percentage of depression of first twitch (T1) was used as the study parameter. The dose response relations of vecuronium and atracurium were determined using the cumulative dose-response technique. RESULTS: The cumulative dose-response curve of vecuronium was shifted to the left in a parallel fashion compared with that of atracurium. As assessed by linear regression, the potency ratio of vecuronium:atracurium was 6:1. There were significant differences in the ED50, ED90, and ED95 between the two drugs. After the intravenous administration of equipotent doses of both drugs (1.5 x ED95), the duration of peak effect, clinical duration, recovery index, and total duration were not significantly different between the two drugs. CONCLUSIONS: The neuromuscular blocking potency of atracurium is approximately 17% that of vecuronium in anesthetized Chinese adult patients. Following equipotent doses, the time course of action of atracurium is similar to that of vecuronium. PMID- 17424754 TI - Ultrasonographic examination to search out the optimal upper arm position for coracoid approach to infraclavicular brachial plexus block--a volunteer study. AB - BACKGROUND: Infraclavicular brachial plexus block has been widely used for surgical procedures below the mid humerus owing to its excellent anesthetic quality and ease of practice. However, what is the optimal upper arm position for carrying out the procedure still lacks consensus of opinion. The primary goal of this study was to determine the optimal upper arm position for coracoid infraclavicular block by ultrasonographic examination. METHODS: High-frequency (5 10 MHz) ultrasonographic examination on the vertical line 2 cm medial to the coracoid process was performed in 40 volunteers. We assessed the influence of four different upper arm positions on the topographic anatomy of the infraclavicular region. Ultrasonography-derived distances and morphometric measurements were applied to evaluate the optimal puncture site. The deviation of coracoid puncture site from the ultrasonographically modified ideal puncture site in distance was also recorded. RESULTS: When the upper arm was abducted 900, the brachial plexus was much closer to the skin (1.67 cm) and farther from the pleura (1.15 cm) as compared with other positions. In this position, the revealation of anterosuperior plexus relative to artery, identification of all three cords and pleura were 53.8%, 64.1% and 87.2%, respectively. We also found that as the upper arm was drawing from abduction to adduction the ideal puncture site tended to shift more inferiorly. CONCLUSIONS: We recommend the most optimal position for carrying out coracoid infraclavicular brachial plexus block is to abduct the upper arm 90 degrees with external rotation of the shoulder. Though ultrasonographic guidance is suggested for infraclaricular brachial plexus block, an optimal position for puncture site determined by anatomical landmark is also acceptable. PMID- 17424755 TI - Earlier cessation of desflurane supply in closed-circuit anesthesia reduces emergence time in patients undergoing breast surgery. AB - BACKGROUND: Minimizing the time of anesthesia emergence can facilitate faster patient turnover in the operating rooms of a busy surgery center. According to Lin's new concept of inhalation uptake, after turning off the vaporizer under close-circuit anesthesia (CCA) with a very low fresh gas flow rate, the concentration of desflurane decreases at a slow rate. The aim of this study was to determine if earlier cessation of desflurane supply would shorten the emergence time and at the same time register the changes of desflurane concentration in the circuit after turning off the vaporizer. METHODS: 30 patients were randomly assigned to two groups, i.e., the control group and the study group. In the control group, the desflurane supply was continued up till the end of the operation, while in the study group the desflurane supply was cut off prior to the suturing the skin. In the study group, data regarding the hemodynamic changes, time from turning off desflurane with high flow washout to wakefulness, and the inspired as well as the expired desflurane concentrations at the low-flow anesthetic phase were collected. The time required from high flow washout to emergence was recorded in all patients. Inter-group and intra-group data were analyzed with nonparametric 2-independent-samples Mann-Whitney test and 2 related-samples Wilcoxon signed ranks test, respectively. RESULTS: Under CCA with similar surgical duration, the patients in the study group emerged from anesthesia significantly faster than those in the control group (5.6 +/- 1.9 min versus 8.8 +/- 2.3 min; P < 0.05), without molestation of stable hemodynamic signs. At the low-flow wash-in stage, the inspired desflurane concentrations were significantly higher than the expired ones from 0 to 2nd min; no significant difference was noted from the 3rd to 6th min, but after which the expired concentrations were significantly higher. Desflurane concentrations decreased most noticeably during the first 5 min (0.35 +/- 0.14%), and then the decrease was moderating from 6th to 10th min (0.21 +/- 0.58%) and staggered from 11th to 15th (0.14 +/- 0.06%). The mean duration of low flow wash was 25.6 +/- 11.6 min. No patient reported awareness during surgery. CONCLUSIONS: Ceasing desflurane supply earlier in CCA (250 mL/min) significantly shortens emergence time without significant hemodynamic changes. PMID- 17424756 TI - The feasibility of surface landmark for coracoid infraclavicular brachial plexus block by ultrasonographic assessment. AB - BACKGROUND: The coracoid infraclavicular block first introduced by Whiffler provides a safer and easily approach than classic infraclavicular block. In this technique, the anatomy-based puncture site is 2 cm medial and 2 cm caudal from the coracoid process. This prospective study was purposed to evaluate the feasibility of surface landmark-based coracoid block by ultrasonography. METHODS: High-frequency ultrasonographic examination was performed in 80 volunteers along the vertical line 2 cm medial to the coracoid process. The C point (C) is defined as landmark-based puncture site. The U point (U) is defined as the ultrasonographically modified optimal puncture site. After identifying the neurovascular bundle, the extent of precision based on landmark was examined and ultrasonographic measurements were also done. Demographic data was applied to correlate with the deviation between C and U. RESULTS: The landmark-based puncture site for coracoid infraclavicular block was found to have a fair precision rate of 74.4%, although not high enough to provide a reliable puncture in daily practice. There was a significant trend toward a more superior puncture site of 2.95 mm (95% CI, 1.2-4.7). In female subjects, U was 5.12 mm (95% CI, 2.91-7.33) superior to C which was statistically significantly (P < 0.001). In male subjects, U was not significantly superior to C. CONCLUSIONS: Ultrasonographic guidance is suggested whenever anatomical precision is inadequate or meeting with great individual bodily variance which renders landmark-based technique difficult. However, if this facility is not available, the gender discrepancy in measurement should be seriously considered when coracoid process is used as the landmark. PMID- 17424757 TI - Persistent systemic air embolism and delayed pulmonary hemorrhage after weaning from cardiopulmonary bypass--a case report. AB - Systemic air embolism (SAE) occurring during cardiac surgery is usually associated with high morbidity and mortality. We present a fatal case of persistent SAE identified by transesophageal echocardiography (TEE) after weaning from cardiopulmonary bypass (CPB). Perplexities in identification of a bronchovascular fistula and hesitation in aggressive management to arrest the resultant continuous air entry into systemic circulation caused death as an aftermath. Related instances in literature have been reviewed and appropriate managements are herein discussed. PMID- 17424758 TI - Successful management of severe upper airway obstruction during emergence of anesthesia in consequence of fracture of deflated laryngeal mask airway due to biting--a case report. AB - Biting the laryngeal mask airway during general anesthesia in the absence of a bite block as a forestallment is a common but usually uncomplicated event. We report a young healthy adult female patient who underwent removal of fixation implant in the right elbow under general anesthesia, during the emergence of which she bit and severed the airway tube of the laryngeal mask airway (LMA) after cuff deflation and developed upper airway obstruction in consequence of air blockade by the displaced deflated LMA cuff remaining inside the mouth. Removal of residual part of the LMA in the mouth was successful with propofol re anesthetization without molestation of 02 saturation. We discuss the management of this critical airway condition resulting from fracture of deflated LMA in the closed mouth. PMID- 17424759 TI - Postoperative delayed respiratory failure caused by Guillain-Barre syndrome--a case report. AB - Postoperative respiratory failure caused by Guillain-Barre syndrome (GBS) is a rare complication after general anesthesia. We report a GBS patient who after receiving an operation for polycystic liver disease under general anesthesia developed weakness of upper and lower extremities on the 3rd postoperative day, with decreased deep tendon reflex, which ultimately evolved into respiratory failure. Slurred speech and bilateral ptosis were also noted. All these manifested an acute peripheral polyneuropathy combined with bulbar involvement. According to the clinical picture, CSF examination and results of nerve conduction tests, Guillain-Barre syndrome was diagnosed. Plasmapheresis was immediately arranged and her motor weakness problem was soon improved after treatment. No neurological sequelae were found two months after discharge. Postoperative muscle weakness is usually caused by residual effects of anesthetic agents or surgical complications. But it may sometimes be related to some rare neurological diseases. To prompt an accurate diagnosis of Guillain-Barre syndrome is important because it can help forestall fatal complications. In addition, the prognosis will be encouraging with early treatment. PMID- 17424760 TI - Gabapentin relieves post-dural puncture headache--a report of two cases. AB - Post-dural puncture headache (PDPH) is a vexing problem of the patients following spinal anesthesia or a complication relative to inadvertent dural puncture in epidural anesthesia. The prevention and management of PDPH contain a laddered forestallment and therapy with varying results. The anticonvulsant gabapentin has been reported to be effective in prophylaxis and treatment of headaches. We report here two cases of PDPH, who failed to respond to traditional analgesics, but had good response to gabapentin. After treatment with gabapentin 400 mg three times daily, the headache was relieved remarkably in 24 hr. Discussions of the pathophysiology of PDPH, pharmacological actions of gabapentin, and possible mechanisms of action of gabapentin on PDPH are brought forward in the text. PMID- 17424761 TI - Birth asphyxia during general anesthesia for a morbidly obese parturient with gestational diabetes mellitus undergoing cesarean section--a case report. AB - A 26-year-old pregnant woman with a body mass index of 60 was scheduled for cesarean section at 36 weeks' gestation. She was found having affected with gestational diabetes mellitus and hypertension at 17 weeks of pregnancy. The blood glucose level as controlled by subcutaneous injection of insulin was maintained at the level of 110-140 mg/dL. Hypertension was also controlled by methyldopa and hydralazine with the systolic pressure maintaining at 140-180 mmHg during the pregnancy. Abnormal perfusion of umbilical artery without compromise of placental function was found twice by Doppler prenatal examination at gestation of 33 and 34 weeks respectively. The operation was performed under general anesthesia. However, apnea and low Apgar score of the neonate were noted. General anesthesia was thought as the root cause of this event at first. After a series of examination and management, hypoglycemia and uteroplacental perfusion insufficiency were considered as the causes of this morbidity. The neonate's condition was improved in 48 hr after close care. PMID- 17424762 TI - Subglottic stenosis after thoracotomy--a case report. AB - Traumatic intubation, reintubation, intubation with endotracheal tube of inappropriate size, and failure to firmly secure the tube may contribute to the development of subglottic stenosis. Systemic factors such as sepsis, hypotension, autoimmune and granulomatous disorders have all been implicated as contributing causes. In addition, a risky circumstance that might be considered important in the development of airway damage is the occurrence of gastreoesophageal reflux (GER), particularly in thoracotomy operations, where the patients are placed in the lateral position. The purpose of this report is to describe a patient who developed subglottic stenosis following a thoracotomy. The possible causes are macrotrauma due to multiple intubations and microtrauma due to inappropriate tube size in the course of anesthesia. Furthermore, GER may worsen mucosal injuries, which may be precipitated by the lateral position. PMID- 17424763 TI - [The scientific basis for healthy eating]. AB - A diet is healthy when it encourages good health and reduces the risk of diet related chronic diseases. Epidemiological research has demonstrated a close relationship between diet and the risk of developing these chronic diseases, which are characterized by high morbidity and mortality. The Mediterranean diet is a tradition that goes back for centuries and which contributes to excellent nutritional status; this diet is highly palatable and forms part of world culture. Some of the most outstanding recent studies on the relationship between unhealthy lifestyle and the risk of disease are reviewed. PMID- 17424764 TI - [Care of patients who have undergone bariatric surgery]. AB - A wide range of problems are caused by obesity, since patients are affected not only physically, but also psychologically, socially and in their working life, with major socio-economic consequences. The application of technical surgical has opened a new field in the treatment of this complex illness, long term treatment being the only effective kind for morbid obesity. With most surgical techniques, the patient should control food intake for life and maintain some adapted habits of physical activity. For this reason it is necessary for professionals in different areas to help patients to develop strategies to persevere with their programs. Evaluation of the patient's physical and psychological needs should be carried out prior to the planning of appropriate measures. Outpatient follow-up must focus on educating, motivating, training and advising the patient nutritionally, so that he or she is able to modify his or her eating habits and maintain a healthy lifestyle during the pre- and postoperative phases. Evaluation using Patterns serves as a tool to analyze disorders in the patient. Once the situation has been evaluated, we can proceed to the nursing diagnoses (NIC) on which we will base the interventions and pertinent activities which, in combination with those carried out by other members of the multidisciplinary team, will achieve optimum results. With all the information obtained, properly organized and discussed by the team, it is possible to personalize the Planning of Care with food registration, food choice, elaboration of a weekly diet, and the general dietary recommendations. Each of the results obtained (NOC) is evaluated in terms of the possible indicators, on the corresponding scale of measurement. In addition to patient motivation, effective instructions about changes in lifestyle and the need to accept treatment and follow-up are the key to positive collaboration with the multi-professional team in the application of strategies. PMID- 17424765 TI - [A comprehensive approach to treating adolescent obesity]. AB - Obesity started to appear as a public health problem at the end of the 20th century; however, it is currently acquiring epidemic dimensions. In most cases this pathology is the result of inappropriate eating habits, which are becoming a major dietary, psychological and social problem in developed countries. In fact, the most serious consequences of obesity in adolescence appear in adulthood. Eating behavior should be conceived in an all-round perspective, including the interaction of the characteristics of the subject (biological, psychological, cultural), with the food the subject can easily access and the environment around him/her (especially peer-group and advertising pressure). In view of these conditions, it remains clear that the treatment of these patients should be individual and integral. In order to obtain a good prognosis in obese adolescents, not just weight loss through restricting their diet, an increase in physical activity should be encouraged, as well as a behavioral change affecting whole lifestyle habits. To this end, an adequate psychological monitoring is essential. PMID- 17424766 TI - [Peripheral parenteral nutrition]. AB - The Target of this review is to provide an up-to-date overview of the advantages and disadvantages of Peripheral Parenteral Nutrition (PPN), including techniques, indications and results. The new generation catheters, together with a better knowledge of the intermediary metabolism, give us the opportunity to use PPN in many clinical situations for short periods of time. PPN is an alternative to Total Parenteral Nutrition (TPN) and is also a complement to enteral nutrition and to the oral route. Progress in the design of catheters and their materials, in infusion techniques and better knowledge of optimal nutrients has made PPN into an effective, safe and useful therapy in the treatment of patients for certain periods of time. PMID- 17424767 TI - [Obesity and functional foods: are the new ingredients and products effective?]. AB - According to WHO data, obesity and overweight have reached the level of a world wide epidemic: more than one billion adults are overweight, and at least three hundred million of them are obese. Obesity is associated with the main chronic diseases of our time. Obesity can reduce the life expectancy by 10 years and in Spain it is calculated that the health care costs account for 7% of the total. In the Spanish adult population, 14.5% are obese, and 38.5% are overweight. In the fight against obesity, the food industry has been incorporating new ingredients intended to help consumers obtain a suitable weight. In this study, we review the scientific evidence about some products and ingredients, discussing the fact that the main problem is their scant scientific support. Advertisers make much of supposed benefits which often lack a scientific basis, as in the case of products with a supposed effect on thermogenesis, metabolism and oxidation of fats, products which make people feel full, products affecting fat absorption, diuretics, laxatived, sedatives and substances affecting the central nervous system. PMID- 17424768 TI - [Community nutrition]. AB - A growing body of scientific and epidemiological evidence indicates that diet and health are related: diet may be a risk factor or have potential protective effects. As a consequence, the focus of nutrition research has experienced a shift towards qualitative aspects of diet which could influence chronic disease, longevity, quality of life and physical and cognitive performance, leading to the development of Community Nutrition. The main undertakings in a Community Nutrition Unit are related to the identification, assessment and monitoring of nutrition problems at the community level and to planning, design, implementation and evaluation of nutrition intervention programs. Such programs combine a number of suitable strategies in a whole population approach, a high risk approach or an approach targeted at specific population groups, and are implemented in different settings, such as the work place, schools or community organizations. Community nutrition interventions aim to gradually achieve change in eating patterns towards a healthier profile. Community Nutrition programs require the use of a combination of strategies and a working group of people from different backgrounds. Many factors influence the nutritional status of an individual or a population. In order to gain effective work output, sound understanding of these patterns and a practical surveillance system are required. PMID- 17424769 TI - [Dietary interview: a useful tool for data collection]. AB - Dietary interview is a very useful tool in the process of evaluating the nutritional state of a person and, of course, in the process of dietary counseling, particularly in the case of patients suffering from chronic conditions. The complexity of gathering information at individual or population level requires different models of questionnaires and tools to enhance the identification of the data collected. The current paper is based on the consensus document about dietary interview, made by a working group created during the 1st Meeting of the Spanish Dietitians' and Nutritionists' Association held in Barcelona 2002. PMID- 17424770 TI - [Prevention of childhood osteoporosis]. AB - Bone mineralization depends on genetic, nutritional, endocrine, metabolic and mechanical factors. Dairy products form the major source of dietary calcium and vitamin D, both of which are essential for optimal bone mineralization. Osteoporosis is defined as a reduction in bone mass; it is exacerbated by malnutrition, low weight, poor intake of vitamin D and calcium and lack of physical exercise. The purpose of this article is to encourage pediatricians to make an effort about the prevention of childhood osteoporosis. PMID- 17424771 TI - [New challenges in basic and applied nutrition]. AB - The quality standards for nutrition and food have undergone major changes during recent years in parallel with the increase in scientific knowledge in the area of food and health. Trends in consumer demands are changing accordingly. While during the last century our concerns were firstly focused on ensuring the availability of basic foods and later on ensuring its safety, nowadays our society is concentrating on improving wellbeing and, particularly, on tackling and preventing the major chronic diseases such as cardiovascular diseases, diabetes, obesity, diverse types of cancer, osteoporosis and autoimmune diseases. These problems account for almost 50% of the diseases and 60% of deaths over the world, and are directly related with the food we eat, irrespective of the importance of other factors (age, sex, physical exercise, genetic predisposition, nicotine poisoning or alcoholism). Europe has decided to go ahead with a major legislative change in the food sector (the Regulation on nutrition and health claims made on foods) which opens new perspectives to improve public health and offer clear economic growth for the health-related food sector. Putting into practice this new legislation and its consequences constitutes a major challenge. Nevertheless, new challenges are foreseen: our response to food depends on our individual genetic characteristics (nutrigenetics); it also depends on the individual history that is being imprinted (in a permanent or temporary form) on our chromosomes (epigenetics), as determined by the individual life style (food, different episodes and facts, including emotions) and, particularly, nutrition during the most active stages of our development. For the future, the additional contribution we will ask of food is to promote health and well-being in all facets. The requirement is that it should develop in a free framework, based on the best scientific available advice, with transparency as a fundamental guarantee. In this context, new disciplines (e.g. nutrigenomics) are emerging and new trends towards a more conscious, better informed, consumer are foreseen that will allow an appropriate reaction to these new challenges in the medium and long term. PMID- 17424772 TI - [Innovations in food processing: nonthermal methods]. AB - Consumers' desires for foods that are minimally preserved and processed are encouraging the development of new methods for the inactivation of micro organisms in foods. Non-thermal processes (such as high hydrostatic pressure, ultrasound, irradiation, high intensity pulsed electric fields, oscillating magnetic fields and light pulses) can be used for the inactivation of food spoilage micro-organisms without affecting the quality of the food. While the efficacy of many of these methods was demonstrated many years ago, technological advances are only now beginning to make possible their commercial exploitation. PMID- 17424773 TI - Neurocops: the politics of prohibtion and the future of enforcing social policy from inside the body. AB - Over the next decade an increasing number of new "pharmacotherapy" medications will become available with the potential to tremendously impact the use and abuse of illegal drugs and the overall direction of national and international drug policy. These pharmacotherapy medications are designed to block or significantly reduce the "highs" elicited by illegal drugs. Used as part of a drug treatment program, pharmacotherapy medications may provide valuable assistance for people voluntarily seeking a chemical aid in limiting or eliminating problem drug use. However, the tremendously politicized nature of the "drug war" raises substantial concerns that, in addition to those who voluntarily choose to use such medications, some people will be compelled to use them. This article concludes that in the absence of extraordinary circumstances, governmental action forcing or coercing a person to use a pharmacotherapy drug would violate a number of important legal rights. Among the rights implicated by compulsory use of pharmacotherapy drugs are the right to informed consent, the right to bodily integrity and privacy, the protection against cruel and unusual punishment, and the right to freedom of thought. PMID- 17424774 TI - Pregnant women inmates: evaluating their rights and identifying opportunities for improvements in their treatment. PMID- 17424775 TI - Too much religious freedom? Infants infected with herpes after Jewish mohel applies oral suction to circumcised penises. PMID- 17424776 TI - Ohio's sex offender residency restriction law: does it protect the health and safety of the state's children or falsely make people believe so? PMID- 17424777 TI - One and done: how Ohio's one-year, nonrenewable visiting medical faculty certificate is harming the state' s economic recovery. PMID- 17424778 TI - The current state of advance directive law in Ohio: more protective of provider liability than patient rights. PMID- 17424779 TI - Gene expression profiling in bone tissue of osteoporotic mice. AB - Ovaricetomized (OVX) animals represent an optimal model to investigate bone loss in osteoporosis. To further elucidate the underlying mechanisms of decreased bone formation and increased bone resorption following OVX, we conducted gene expression profiling experiments using bone samples of ovariectomized C57BL/6J mice. Following OVX, genes involved in immune response, cell cycle regulation, growth, apoptosis and bone resorption were upregulated, while genes that are important for regular cell processes, mitosis, metabolism of carbohydrates, extracellular matrix structure, angiogenesis, skeletal development and morphogenesis were downregulated. Among bone specific genes we observed upregulation of interleukin 7 (IL-7), IL-7 receptor and matrix metallopeptidase 8, while genes such as transforming growth factor-beta 3, procollagen type I and procollagen type VI exhibited marked decrease in expression. We also observed downregulation of two genes, parathyroid hormone receptor 1 and WD repeat domain 5, that are involved in skeletal development but were not previously reported to be altered in osteoporosis. We further performed gene set enrichment analysis (GSEA) in order to calculate enrichment of pathways specifically altered in murine bones following ovariectomy. In conclusion, OVX greatly influences expression of various genes involved in diverse biological processes confirming the notion that numerous pathways play an important role in pathophysiology of osteoporosis. PMID- 17424780 TI - [Epidemiology of osteoporosis]. AB - In many countries, including Croatia, there is no disease registry for osteoporosis and osteoporotic fractures. Epidemiological data show that the prevalence of osteoporosis is much higher in Asian people than in white European or North American populations, where the prevalence ranges between 10% and 15%. Epidemiological characteristics of osteoporosis in Croatia are similar to other European countries, though the incidence of osteoporotic fractures is somewhat higher. According to the annual report of registered diseases, the number of patients with osteoporosis and osteoporotic fractures increased during the last 10 years. In 2004, 0.70 % of all registered diseases in Croatia referred to osteoporosis and osteomalacia. The most important risk factors for osteoporosis are age, heredity, lifestyle, chronic diseases, hormonal abnormalities and physical characteristics of bone. Like in other chronic diseases, prevention measures are most important for disease control. PMID- 17424781 TI - [Glucocorticoid-induced osteoporosis]. AB - Glucocorticoids are the most common cause of drug-induced osteoporosis. Given the widespread use of oral glucocorticoids in the treatment of autoimmune, pulmonary, gastrointestinal disorders and organ transplantation, attention to glucocorticoid induced osteoporosis has substantially increased. Bone loss occurs rapidly in the first few months of glucocorticoid therapy. Trabecular bone is affected more than cortical bone. Glucocorticoid treatment is associated with a substantially increased risk of fractures, particularly hip and vertebral fractures. The skeletal effects of glucocorticoids are both dose- and duration-dependent. The patophysiology of glucocorticoid-induced osteoporosis is a complex process, several mechanisms are proposed but not yet fully highlighted. Despite several evidence-based guidelines for the prevention and treatment of glucocorticoid induced osteoporosis and the availability of effective therapeutic options, the proportion of individuals with appropriate evaluation and treatment remains relatively low. PMID- 17424782 TI - [Osteoporosis in men]. AB - With the prolongation of life expectancy, osteoporosis has become an increasing problem in the majority of developed countries worldwide. The paper discusses the frequency, pathogenesis, diagnostic criteria and treatment options for osteoporosis in men. Every third hip fracture occurs in men, and more than 11% of the male population over the age of 50 years suffer the fracture. Diagnostic tests for idiopathic osteoporosis are performed in men under 60 years of age without other potential risk factors of developing the disease. In the majority of cases, their low bone mineral density (BMD) is caused by a low peak bone mass. Secondary osteoporosis occurs in about 30 % of men, and involutionary osteoporosis developed in men over 60 years of age results from their decreased testosterone and IGF-1 levels. The study results showing that BMD levels in both sexes provide similar fracture risk information suggest that the existing diagnostic criteria for female osteoporosis can also be employed in men. It has been proved that biphosphonate and teriparitide therapy significantly increase BMD levels in men. The administration of androgens has been shown to be effective in men with hypogonadism, although their validity for patients with eugonadism has not yet been discussed. An improved knowledge of the bone metabolism and bone remodelling has recently opened the door to an extensive series of molecules that may play a key role in the treatment of male osteoporosis in the future. PMID- 17424783 TI - [Assessment tools in early detection of osteoporosis in dentistry]. AB - Osteoporosis, one of the major skeletal diseases in older age, is characterised by low bone mass and microarchitectural deterioration with a resulting increase in bone fragility and hence susceptibility to fracture. In this review we analyse the systemic and local factors associated with oral bone mass loss. Systemic factors most often correlated with the oral bone mass loss include osteoporosis, renal diseases, hormonal disorders, diet and the impact of different drugs on the bony structure. Chronic periodontal disease, early loss of teeth or the effect of inadequate prosthodontic appliance on the residual ridge are the local factors associated with mandibular bone loss. Different assessment tools for the assessment of mandibular oral bone loss have been proposed, such as DXA absorptiometry, quantitative computed tomography, intraoral microdensitometry, SCORE index and the assessment of the thickness and quality of the mandibular inferior cortical border. Qualitative and quantitative assessment of the mandibular bony structure is of great importance in all fields of dentistry - from periodontology to endodontics and prosthodontics, especially in dental implantology. It is important to make the correct indication prior to dental implant therapy, and taking into account the systemic and local factors mentioned above, assess both the actual quality and quantity of the mandible. PMID- 17424784 TI - [Osteoporotic fractures in the elderly]. AB - The epidemiology of injuries is a significant factor in the structure of mortality and causes of hospitalization of elderly people. Epidemiologically, consequences of injury, i.e. their complications, from reduced mobility, sores, contractures, infections, all over to mortality from hypostatic pneumonia, are strongly emphasized in the elderly. In 2005, more than 17.0% of Croatian population were older than 65 years of age, and 27.0% of the population were over 60. Osteoporotic bone fractures are frequent among the elderly; 5,489 hip fracture cases were registered in Croatia during 2005, and 382 of them died from fracture complications. In total, 97.38% of the dead patients were over 65 years of age. Besides hip fractures, other typical osteoporotic fractures are fractures of vertebral bodies, distal part of the radius, humerus, pelvic bones, etc. Regardless of conservative or surgical treatment for osteoporotic fracture, it is essential to stress out that after the care of fractured bone has been provided, appropriate diagnostic examinations and pharmacological treatment of osteoporosis should also be done. Orthopedic and traumatic surgeons all over the world have to understand that conservative or surgical treatment of osteoporotic fracture of any localization is just one link in this complex chain of managing the disease - osteoporotic treatment. PMID- 17424785 TI - [Treatment of osteoporotic fractures of the spine]. AB - Osteoporosis is a skeletal disease characterized by reduced bone strength due to the loss of bone mass, structural disproportion and reduced calcium level as well as changes in the bone structure caused by trabecular deficit. The loss of bone mass in the vertebrae leads to an increased risk of fractures. Osteoporotic vertebral fractures are two to three times more frequent than hip fractures, especially in women. Osteoporotic compressive fractures of the spine mostly occur in the middle and lower thoracic spine and in the thoracolumbar spine. Common consequences of fractures include long-term pains, decrease in the height and spinal deformity, which significantly correlate with disorders in the gait and mobility, reduced pulmonary function, loss of appetite and in the end increased mortality. Clinical picture in acute conditions manifests itself as a sudden pain in the back following minimal trauma but also often without any trauma at all. Diagnostics is based on a detailed medical history and a thorough clinical examination. Radiological diagnosis relies on standard x-rays. In case of clinical indication (differential diagnosis of tumor or spondylitis, neurological weakness) CT and/or MR imaging is performed. The approach to the treatment of osteoporotic fractures of the spine is individual and is based upon the medical history, localization of symptoms and correlation of the clinical examination with the radiological finding. Open reduction, decompression and fixation are indicated in patients with posttraumatic stenosis and neurological weakness. Current operative methods include vertebroplasty and kyphoplasty, which significantly reduce pain and improve mobility of the patient and consequently, the quality of life. PMID- 17424786 TI - [Treatment of osteoporosis]. AB - Osteoporosis is among the most frequent metabolic diseases affecting 8% to 10% of the population. Since the most disturbing outcome of osteoporosis is a fracture, it is important to identify patients at risk and intervene with pharmacologic therapies and lifestyle changes. Several drugs have shown their ability to reduce vertebral and/or peripheral fractures in patients with osteoporosis. Antiresorptive agents are a basis of therapy, but anabolic drugs have recently widened therapeutic options. Antiresorptive medications, estrogens, selective estrogen receptor modulators, bisphosphonates and calcitonins, work by reducing the rates of bone remodeling. Parathyroid hormone stimulates new bone formation, repairing architectural defects and improving bone density. Strontium ranelate reduces the risk for osteoporotic fractures by both inhibiting bone resorption and increasing bone formation. Other potential therapies for osteoporosis are also reviewed in this article. PMID- 17424787 TI - Report of the Poison Control Centre for the period 1 August-31 December 2006. PMID- 17424788 TI - Institute for Medical Research and Occupational Health, activity report for 2006. PMID- 17424789 TI - Securing the future of critical care nursing in Australia. PMID- 17424790 TI - Managing diarrhoea in intensive care. AB - OBJECTIVE: To assess the incidence of diarrhoea in intensive care patients with a length of stay (LOS) greater than 3 days who were receiving any type of enteral tube feeding, and to measure the effect of implementing a bowel management protocol. DESIGN: A 2-year prospective audit, with an intervention after 12 months. Diarrhoea was defined as bowel activity exceeding three stools of any consistency per day, or three or more unformed stools (or 300 mL) per day, for two consecutive days. SETTING: A tertiary referral intensive care unit (ICU) in a large public hospital. SAMPLE: Six hundred fifty-six consecutive patients admitted to ICU with a LOS >3 days. INTERVENTION: A bowel management protocol was implemented to address both diarrhoea and constipation. MAIN OUTCOME MEASURES: Number of patients who experienced diarrhoea during their ICU stay; number of ICU patient-days on which diarrhoea occurred. RESULTS: After the protocol was implemented, diarrhoea was experienced by 13% fewer patients (p = 0.0002) and occurred on 8% fewer ICU days (p < 0.0001). CONCLUSION: Use of an evidence-based protocol, and improved monitoring and reporting of bowel activity, can decrease the incidence of diarrhoea in ICU patients. PMID- 17424791 TI - Anticoagulation in CRRT: agents and strategies in Australian ICUs. AB - BACKGROUND: Continuous Renal Replacement Therapy (CRRT) should ideally operate with as little interruption as possible. The majority of circuit terminations occur due to clotting. The Longevity of CRRT is able to be improved when the extracorporeal circuit is anticoagulated. AIMS: This article willt focus attention on anticoagulant agents used in Australian intensive care units (ICU) to prevent clotting in the CRRT circuit. DISCUSSION: Anticoagulants reviewed include unfractionated or standard heparin, regional heparinisation, low molecular weight heparins and heparinoids, regional citrate, platelet-inhibiting agents (prostacyclin), thrombin antagonists (recombinant hirudin) and therapy with no anticoagulant use. Each type of anticoagulant was reviewed for mode of action, the method of delivery and how the effect is monitored. Circuit life and the incidence of bleeding were considered as the principle end points in selecting therapy, as well as side-effects with administration such as metabolic disturbances, contraindications to use including allergy and ease of use in the clinical environment. CONCLUSION: No approach to anticoagulation has yet been reported to be as successful in extending circuit life, whilst remaining inexpensive, easy to manage and easy to reverse, as unfractionated heparin. Certain patient conditions may preclude the use of heparin, such as heparin induced thrombocytopenia (HIT); then heparinoids, thrombin antagonists and sodium citrate are suggested as alternatives. Regional citrate reduces haemorrhagic complications in patients who have coagulation disorders or are at risk of bleeding. Clinical experience with various agents and strategies should also influence choice. The option of no anticoagulant may be appropriate in selected patients rather than more expensive and less familiar drugs. PMID- 17424792 TI - Chest x-ray quiz. Why would this patient be breathless? Large left pleural effusion compressing, and hence collapsing the left lung, with more blood being delivered into the right pulmonary circulation. PMID- 17424793 TI - Comparison of dynamic measurements of pulse contour with pulsed heat continuous cardiac output in postoperative cardiac surgical patients. AB - Cardiac output (CO) can be measured using bolus thermodilution via a pulmonary artery catheter (PAC) and as continuous cardiac output (CCO), using pulsed heat thermoditution. Pulse contour cardiac output (PCCO) measures continuous CO by analysis of the arterial waveform after calibration with thermodilution CO. The Pulsion Medical Systems (PiCCO system) achieves this by transpulmonary aortic thermodilution (TDtpa). There is uncertainty regarding the agreement between TDtpa, CCO, and PCCO CO measurements in situations of rapid haemodynamic changes. We studied the agreement of the measures by comparing digital recordings of cardiac index (CI) determined by PCCO and CCO (PCCI and CCI, respectively) made during periods of haemodynamic instability. After ethics committee approval we studied four post-coronary artery bypass graft patients, in the immediate postoperative period. Each patient had a 7.5F CCO catheter (Edwards Lifesciences) and a 5F, 20cm PCCO femoral artery catheter. Digital recordings were obtained for the first 12-18 postoperative hours. Six epochs of instability were identified in the first two to three postoperative hours, and at the commencement of inotropic or vasoactive drugs. Notable features, despite frequent PCCO calibrations, were the marked difference of PCCI compared to CCI. In contradistinction, they tracked very closely during a period of stability. Limitations of both methods were noted. Whilst PCCO responded to rapid change, it developed significant error during haemodynmamic instability and requires frequent recalibration. CCO on the other hand has a considerable time lag in responding to changes in CO. The way a monitor measures CO must be taken into account when using the data in clinical management. PMID- 17424794 TI - [The health network, the future of internal medicine? A California example]. PMID- 17424795 TI - [Narcolepsy: current view]. AB - Narcolepsy is a primary sleep disorder which may or may not arise associated with catalepsy. It is a relatively frequent disease with 2 peaks of appearance in childhood and in young adulthood. It has genetic components which do not seem sufficient for its demonstration. It often depends on the environment and its clinical picture can be very variable, particularly in children. It is thus under diagnosed and often confused with other diseases. In this short review of the literature we discuss: narcolepsy's history, clinical evaluation, diagnoses criteria, patient's history, clinical status and complementary examinations, clinical picture in the child and in the teenager, epidemiology, neurobiology, differential diagnoses, symptomatic narcolepsy, current treatments and internet sites of reference. PMID- 17424796 TI - [The role of steroids in the treatment of COPD]. AB - The COPD is a problem of public health. The assumption of responsibility of the COPD depends on its severity. The drugs most often used are the bronchodilators of short and long half-life. The combination of tiotropium and beta-2 agonist of long half-life brings small advantages of quality of life without influencing the evolution of the disease. The systemic steroids are to be reserved for severe exacerbations, and their use must be limited to two weeks. The inhaled steroids as basic treatment are to be reserved for the most severe cases. Several new molecules are being studied in the treatment of the COPD, for which the fight for the final suspension of the tobacco use is one of the determining elements. PMID- 17424797 TI - [Respiratory insufficiency]. AB - Respiratory failure results from the incompetence of the respiratory system to eliminate the CO2 produced by the metabolism and to extract the O2 from the atmosphere. Manifestations of the chronic form are: dyspnea, fatigue, headaches, oedema, polyglobulia, and also anxiety, denutrition, social isolation. If neglected, it will lead to pulmonary hypertension and rapidly fatal chronic cor pulmonale. At home mechanical ventilation and oxygen-therapy are the basis of treatment. Major goals of chronic treatment are: control of dyspnea and anxiety, maintenance of a regular physical activity and a sufficient alimentation. These goals can only be achieved by a network of professionals taking charge of these complex patients, with their periodic inclusion in a pulmonary rehabilitation program. PMID- 17424798 TI - [Pulmonary blastoma in adults]. AB - Pneumoblastoma (PB) is a rare, malignant, primary, pulmonary tumour, of young adults. Its discovery is fortuitous in asymptomatic patients. It is a well delimited, homogeneous lesion in the lung periphery. Histologically, its structure looks like a normal fetal lung. The surgical resection is the treatment of choice. The radiotherapy is an empirical palliative treatment to relieve dyspnea when other treatments failed. The prognosis is bad: 16% survive 5 years and 8% beyond 10 years, all treatments included. Metastases could appear in the liver, the brain and the bone. PMID- 17424799 TI - [Bronchiectasis: a reemerging disease]. AB - Bronchiectasis has a barely studied and under-estimated prevalence. Diagnosis is based on clinical history and CT scan. It is necessary to make some tests to clarify its etiology. Microbiology of expectoration, physiotherapy, prolonged and repeated antibiotherapy play the key role in treatment. We lack studies to recommend other therapies. Surgery is reserved for localized lesions. Ten years mortality is 30%, but in ICU it is very high. An early and aggressive treatment seems justified. PMID- 17424800 TI - [A large network of integrated medical care: the example of Kaiser Permanente in California. First-hand report by a physician]. AB - The Kaiser Permanente (KP) system of integrated medical care is a unique model of medical organization in the USA which achieves the twin goals of economic efficiency and first-rate care. Organizationally, it is quite different from most health maintenance organizations (HMOs). The doctors remain independent, but in an exclusive marriage with the Kaiser Hospitals and the Kaiser insurance, both of which are non-profit. KP cares for over 8 million members. KP ensures continuity of patient care whether at home, as an outpatient, or when hospitalized, and promotes prevention among healthy members. The integration of all services produces very high indices measuring quality of care, as investigated by both the press and official government agencies at a surprising low cost. The system also was found to be more cost-effective than the National Health System in the United Kingdom. PMID- 17424801 TI - [Body organ traffic and illegal transplantations: state of ethics and responsibilities]. PMID- 17424802 TI - [Reflections after the December Medical Meeting...]. PMID- 17424803 TI - [Unique case]. PMID- 17424804 TI - [One in two of the French does not make the difference between embryo and fetus]. PMID- 17424805 TI - [Red alert on "Viagra for females"]. PMID- 17424806 TI - [France and the neo-eugenics]. PMID- 17424807 TI - [Isolated small totalitarianism ]. PMID- 17424809 TI - Old faces, new jobs, tough choices. PMID- 17424808 TI - Health care at your fingertips. PMID- 17424810 TI - Law offers expanded opportunities on HSAs. PMID- 17424811 TI - Universal coverage proposals call for improved care. PMID- 17424812 TI - You can take it with you. PMID- 17424813 TI - Uncommon challenges. PMID- 17424814 TI - The road to recovery. PMID- 17424815 TI - Narrowing the gap. PMID- 17424816 TI - Employers to health plans: more information, please. PMID- 17424817 TI - Understanding universal health accounts. PMID- 17424818 TI - Selective sourcing for the agile enterprise. PMID- 17424819 TI - Web sight: the clinical vision for payer-provider portals. PMID- 17424820 TI - Living with cancer. PMID- 17424821 TI - It's universal. The democrats are all thinking big on fixing health care. How an untouchable issue became the hot topic for 2008. PMID- 17424822 TI - What now? Our feverish planet badly needs a cure. PMID- 17424823 TI - Parents: relax. Teens are acting more responsibly. It may even be time to reward them with some of the rights adults have. PMID- 17424824 TI - Chasing paper from medicine. PMID- 17424825 TI - Knee deep. PMID- 17424827 TI - Clearing of suspensions of Micrococcus lysodeikticus catalysed by lysozymes from hen, goose, and turkey egg whites, human milk, and phage T4. Assessment of potential as signal generators for homogeneous enzyme immunoassays for urinary steroids. AB - Lysozymes (3.2.1.17) from goose (Anser anser) egg white, turkey (Melagris gallopavo) egg white, phage T4 and human milk were compared with hen egg white lysozyme in their ability to clear a suspension of Micrococcus lysodeikticus. All of the lysozymes, except hen egg white lysozyme, catalysed the clearing of the Micrococcus lysodeikticus suspension in a biphasic fashion. Compared to hen egg white lysozyme, the total absorbance or transmission change over 5 and 20 minutes was less in all cases, except for human lysozyme. Human lysozyme was, therefore, a potential alternative, more rapid signal generator for the measurement of urinary estrone glucuronide excretion rates because of its structural similarity to hen egg white lysozyme. The apparent K(M) values for hen egg white lysozyme increased with the enzyme concentration. PMID- 17424828 TI - Utility of slot-blot-ELISA as a new, fast, and sensitive immunoassay for detection of carcinoembryonic antigen in the urine samples of patients with various gastrointestinal malignancies. AB - Carcinoembryonic antigen (CEA) is the most widely used clinical tumor marker. CEA immunoassay has found acceptance as a diagnostic adjunct in clinical diagnosis of gastrointestinal tumors (GIT). Several immunoassays have been established for detection of CEA in plasma, serum, tissue, feces, and urine of cancer patients using polyclonal or monoclonal antibodies raised against CEA. Some of these assays display both high sensitivity and specificity for the detection of CEA. However, these assays require special and highly expensive equipment and the procedures require long periods for their completion. In the present study, we established a Slot-Blot Enzyme Linked Immunosorbent Assay (SB-ELISA), based on anti-CEA monoclonal antibody (CEA-mAb), as a new, simple, fast, cheap, and non invasive immunodiagnostic technique for detection of CEA in the urine of GIT patients. Urine and serum samples were collected from 248 GIT patients (58 with pancreatic cancer, 20 with hepatoma, 23 with ampullary carcinoma, 15 with hilar cholangiocarcinoma, 28 with gastric cancer, 14 with esophageal cancer, and 90 with colorectal cancer). Moreover, urine and serum samples were collected from 50 healthy individuals to serve as negative controls. The traditional ELISA technique was used for determination of CEA in the sera of GIT patients using anti-CEA monoclonal antibody. A comparison between the results of both techniques (ELISA and SB-ELISA) was carried out. The traditional ELISA detected CEA in the sera of 154 out of 248 GIT patients with a sensitivity of 59.8%, 51.7% positive predictive value (PPV) and 75.37% negative predictive value (NPV). In addition, it identified 15 false positive cases out of 50 healthy individuals with a specificity of 70%. The urinary CEA was identified by a Western blotting technique and CEA-mAb at a molecular mass of 180 Kda. The developed SB-ELISA showed higher sensitivity, specificity, PPV, and NPV (70.1%, 78%, 62.4%, and 82.13%, respectively) for detection of CEA in the urine of GIT patients. The semi quantitative SB-ELISA showed a higher overall efficiency of 72.8% versus 63.4% in the case of the quantitative ELISA, for detection of CEA. In conclusion, SB-ELISA is more efficient for detection of CEA in gastrointestinal tumors. It is a simple, rapid, non-invasive, and sensitive assay. Moreover, all steps of the SB ELISA are performed at room temperature, without the use of expensive equipment; this may enhance the application of this assay in field studies and mass screening programs. PMID- 17424829 TI - Development and validation of a sensitive radioimmunoassay procedure for estimating FSH in mithun (Bos frontalis) plasma. AB - The present study was designed to develop and validate a simple and sensitive radioimmunoassay (RIA) procedure to estimate FSH in mithun (Bosfrontalis) plasma. The assay was carried out in 100 [L of mithun plasma. The bovine FSH standards (10 to 5000 pg/100 microL/tube) in hormone-free plasma were used in the assay. The sensitivity of the assay was 20 pg/100 microL/tube, which corresponded to 0.20 ng/mL plasma. The 50% relative binding sensitivity of the assay was 80 pg/100 microL/tube, which corresponded to 0.80 ng/mL plasma. The intra- and inter assay coefficients of variation were 4.6% and 12.4%, respectively. The biological validation of the assay was carried out in plasma samples that were collected during different stages of the estrous cycle. In the entire estrous cycle, plasma FSH concentration (p < 0.01) attained two peaks (on day 3 to 4 before estrus 5.1 +/- 0.3 ng/mL and on the day of estrus 6.9 +/- 0.2 ng/mL). FSH concentration remained at basal level (1.3 +/- 0.1 to 1.6 +/- 0.2 ng/mL) during day 4 to 16 of the estrous cycle. The concentration of plasma FSH was found to be significantly (p < 0.05) higher (4.9 +/- 0.3 to 6.8 +/- 0.5 ng/mL) until 48 h following the estrus onset. In conclusion, the RIA procedure that was developed in the current study is sufficiently reliable and sensitive to estimate different physiological levels of FSH in mithun plasma. PMID- 17424830 TI - Polyreactivity of monoclonal antibodies produced against thyroid stimulating hormone (hTSH). AB - Monoclonal antibodies against human thyroid stimulating hormone (hTSH) are among the key reagents required in the development of an immunoradiometric assay (IRMA) for hTSH in serum. In this study we have produced and characterized twelve hTSH monoclonal antibodies. Hybridomas were generated by fusion of B-lymphocytes from mice immunized with hTSH and myeloma cells. Clones producing antibodies against hTSH were selected using ELISA (Enzyme Linked Immunosorbent Assay). Antibodies from the selected and cloned hybridoma cells were purified by affinity chromatography and their reactivities were tested by ELISA against a panel of antigens, i.e., hTSH, bovine serum albumin (BSA), and milk protein, and also by studying the binding of these monoclonal antibodies with the radioiodinated antigens (hTSH, BSA, and milk protein). It was observed that some of the hTSH monoclonal antibodies produced were polyreactive, reacting with hTSH as well as with unrelated antigen BSA, while others were monoreactive, reacting only to hTSH. PMID- 17424831 TI - Direct radioimmunoassay for the measurement of serum testosterone using 3H as label. AB - Direct radioimmunoassay (RIA), based on the principle of competitive inhibition for the measurement of serum testosterone, using 3H as label, is described. Testosterone 3-O-carboxymethyloxime-bovine serum albumin (testosterone 3-O-CMO BSA) was used as an immunogen and testosterone, labeled at positions 1, 2, 6, and 7 with 3H was used as tracer. To 12 x 75 mm glass tubes 100 microL of standard (250 to 10,000 pg/mL) and unknown samples were added in duplicate, followed by 100 microL of antibody and 600 microL of tracer (10,000 counts per minute [cpm]) in all the tubes and incubated overnight at 4 degrees C. The bound and free fraction of labeled were separated by adding 200 microL of charcoal followed by centrifugation. The bound radioactivity was measured in the supernatant by using a scintillation fluid containing 2,5-diphenyloxazole (PPO, primary scintillator) and p-bis[2-(5-phenyloxazolyl)]-benzene (POPOP, secondary scintillator). In this new strategy, high ionic strength and low pH of the buffer are utilized to release bound steroid from proteins. The sensitivity of the assay is 270 pg/mL. The analytical recovery ranged from 100.24% and 108.94%. The inter-assay and intra-assay coefficients of variation ranged from 3.38% to 9.56% and 5.69% to 9.84%, respectively. The serum testosterone values obtained by this method were correlated with those obtained by solid phase radioimmunoassay: r = 0.91 (n=34). PMID- 17424832 TI - Direct radioimmunoassay for the measurement of serum progesterone using 3H as a label. AB - Direct radioimmunoassay (RIA), based on the principle of competitive inhibition for the measurement of serum progesterone using 3H as label, is described. Progesterone 3-O-carboxymethyloxime-bovine serum albumin (progesterone 3-O-CMO BSA) was used as an immunogen and progesterone labeled at positions 1, 2, 6, and 7 with 3H was used as tracer. To 12 mm x 75 mm glass tubes, 100 microL of standard (250 pg to 50,000 pg/mL) and unknown samples were added, in duplicate, followed by 100 [LL of antibody and 600 microL of tracer (10,000 counts per minute [cpm]) in all of the tubes and incubated overnight at 4 degrees C. The bound and free fractions of labeled material were separated by adding 200 microL of charcoal followed by centrifugation. The bound radioactivity was measured in the supernatant by using a scintillation fluid containing 2,5-diphenyloxazole (PPO, primary scintillator) and p-Bis[2-(5-phenyloxazolyl)]-benzene (POPOP, secondary scintillator). In the present study, a high ionic strength, along with low and neutral pH of the buffer, is utilized to release bound steroid from proteins. The sensitivity of the assay is 732 pg/mL. The recovery ranged between 94.03% to 100.96%. The inter-assay and intra-assay coefficients of variation ranged from 3.89% to 7.59% and from 9.96% to 12.6%, respectively. The serum progesterone values, obtained by this method, were correlated with those obtained by enzyme linked immunosorbent assay; r = 0.96 (n=94). PMID- 17424833 TI - A sensitive sandwich ELISA for buffalo prolactin. AB - Two murine monoclonal antibodies have been developed against the buffalo prolactin (buPRL). These were designated as 1501 and 1504. Using two MAbs and anti-buPRL rabbit serum, an analysis was performed for the development of a sandwich ELISA (sELISA). The 1504/buPRL/anti-buPRL rabbit serum system was found feasible for sELISA. The sELISA had a sensitivity of 156 pg/mL. The buffalo serum sample showed a parallelism with the standard curve. The intra- and inter-assay coefficients of variance were 8.4% and 9.06%, respectively. These data proved the validity of the assay. PMID- 17424834 TI - Immunodetection of collagen types I, II, III, and IV for differentiation of liver fibrosis stages in patients with chronic HCV. AB - The current study is aimed at evaluating serum collagens and other serum biochemical markers as useful, non-invasive markers of hepatic fibrosis associated with chronic hepatitis C virus (HCV). Collagen types I, II, III, and IV were detected in serum using ELISA and Western blot techniques. The ELISA levels of collagen I, II, III, and IV increased significantly with the progression of fibrosis staging. Based on receiver-operating characteristic (ROC) curve analysis, the collagen type III (70 kDa) and type IV (200 kDa) were more useful than other serum bio-markers for differentiating severe fibrosis from mild fibrosis. Multivariate discriminant analysis (MDA) selected a fibrosis discriminant score (FDS) = [2.345 + Collagen III (microg/mL) x 1.923 + Collagen IV (microg/mL) x 1.544 + ALT (U/mL) x 0.005] - [albumin(g/L) x 0.046]. The FDS correctly classified 87% of the severe fibrosis patients at a cut-off score = 2.2 (i.e., more than 2.2 indicated severe fibrotic liver and less than 2.2 indicated mild fibrotic liver) with specificity of 97%. In a validation study, the FDS was applied to the second cohort of patients and the results were reproduced without significant difference. In conclusion, the developed four-parameter based FDS is useful for identifying severe liver fibrosis in patients with chronic HCV infection. PMID- 17424835 TI - [Excretion of urinary metallothionein and osteal damage induced by cadmium in an environmentally cadmium exposed population]. AB - OBJECTIVE: To observe the relationship between urinary metallothionein excretion and osteal damage induced by cadmium in a general population. METHODS: The inhabitants living in both cadmium polluted and non-polluted areas were asked to participate in this study. Urinary cadmium (UCd) and blood cadmium (BCd) were measured by GF-AAS. Total cadmium(TCd)was evaluated with environmental cadmium exposure. URBP, UB2M, UALB and UMT were measured by ELISA method. UNAG, UNAGB were measured by fluorescence analysis method. Forearm bone mineral density in human were mensurated by SPA. RESULTS: UMT can reflect the change of cadmium body burden. Renal dysfunction and osteoporosis would appear successively after high level of cadmium exposure. UMT had a complex relationship with bone mineral density which related to the amount of UMT excretion. The BMDLs of UCd were calculated using software of BMDS Versionl. 3.2 for these biomarkers. The values of BMDL of these biomarkers were arranged: UNAGB < UNAG < UB2M < UMT < URBP < Tscore < UALB. CONCLUSION: Cadmium exposure could induce bone damage which occurred later than renal dysfunction related to cadmium exposure. UMT could be not only a specific and sensitive biological indicator of cadmium-induced renal dysfunction but also could reflect the damage on bone induced by cadmium. PMID- 17424836 TI - [Effect of dimethoate on dopamines neurotransmitters in brain regions of rat]. AB - OBJECTIVE: To observe changes of dopamine and its metabolite (3,4 hydroxyphenylaceticacid, DOPAC) levels of the brain regions in rats exposed by dimethoate (DM). METHODS: Groups of rats were treated with saline and 38.9, 83.7 and 180.0 mg/kg DM respectively, and were decapitated at different time course from 0.5 to 24 hours after the administration. The dopamine neurotransmitters and its metabolite were determined by reverse-phase high-performance liquid chromatography with electrochemical detection. RESULTS: The concentrations of DA (28% - 122% of control) and DOPAC (89% - 538% of control) varied according with the DM dose and the exposure time. CONCLUSION: These results indicated that DM induced toxic effects could alter the dopamine levels after the different dose and time exposure in the brain regions of rats. PMID- 17424837 TI - [Effect of anatoxin-A on calcium-mediate signal system of brain tissue in mice]. AB - OBJECTIVE: To explore the mechanism of central neurotoxity induced by anatoxin-A (ANTX-A) from the calcium-mediate signal system. METHODS: Mice were administered by i.p. at different doses of ANTX-A for one month, 0.9% of sodium chloride solution was used as the control. Using fluorescence method and colorimetry method, the intracellular calcium concentration and the activity of calcineurin (CaN, PP2B) and ATPase in brain were determined. RESULTS: Calcium concentrations in brain were increased in 12.5, 50 and 200 microg/kg bw groups compared with the control group, the difference had statistical significance (P < 0.01). The activity of PP2B of brain in 200 microg/kg bw group was enhanced compared with the control group (P < 0.05). In all ANTX-A treated groups, the activity of Na+ K+ -ATPase of brain in mice had no obvious changes, but the activity of Ca2+ ATPase in 50 and 200 microg/kg bw groups were significantly reduced compared with the control group ( P < 0.05). CONCLUSION: It is suggested that calcium-mediate signal system could have important regulative action in the central neurotoxity induced by antoxin-A. PMID- 17424838 TI - [Genetic polymorphisms of MPO, NQO1, GSTP1, UGT1A6 associated with susceptibility of chronic benzene poisoning]. AB - OBJECTIVE: To explore the relationship between genetic polymorphisms in MPO, NQO1, GSTP1, UGT1A6 and susceptibility to chronic benzene poisoning (BP). METHODS: A case-control study was conducted. 268 BP patients and 268 workers occupationally exposed to benzene without poisoning manifestations were investigated. The genotype of MPO (rs7208693), NQO1 (rs1800566), GSTP1 ( rs947894 ) and UGT1A6 (rs6759892, rs1105879, rs4124874 ,rs3755319 ,rs887829 and rs4148323) were tested by the TaqMan PCR method. RESULTS: There was a 0.675-fold (95% CI 0.434 - 0.994, P = 0.046) decreased risk of BP in the subjects with GSTP1 rs947894 G alleles compared with those carrying AA genotype. In the subjects carrying allele of MPO rs7208693 A, the risk of BP increased for the individuals carrying allele of UGT1A6 rs6759892 G (OR = 2.702, P = 0.024) compared to those with TT genotype or the individusals carrying allel of UGT1A6 rs1105879 C (OR = 2.619, P = 0.035) compared to those with TT genotype. In drinking population, the individuals carrying the NQO1 rs1800566 TT homozygote genotype had a 9.000-fold (5% CI 1.460 - 55.478, P = 0.021) risk of BP compared to those with CT + CC genotypes. In smoking population, there was 7.000-fold (95% CI 1.555 - 31.575, P = 0.012) of risk in BP subjects carrying NQO1 rs1800566T/T genotype, compared with those carrying CC + CT genotype. Haplotyes analysis of polymorphisms in UGT1A6 showed that compared with those carrying TAATGG haplotype, there was a 1.446-fold (OR = 1.446, 95% CI 1.005 - 2.080, P = 0.046) increased risk of BP for those carrying TACGGG haplotype. CONCLUSION: The subjects carrying allele of MPO rs7208693 A and UGT1A6 rs6759892 G or rs1105879 C at the same time could be more susceptible to BP. The subjects carrying NQO1 rs1800566 TT genotype and together with the habit of smoking or drinking could be more susceptible to BP. Those subjects carrying TAATGG haplotype of UGT1A6 could increase the risk of BP. Further studies should be needed on the association between the genetic polymorphisms in GSTP1 and the risk of BP. PMID- 17424839 TI - [Experssion of human glutathione-S-transferase A1 in CHO cells]. AB - OBJECTIVE: To establish the Chinese Hamster Ovary (CHO) expression system for the human glutathione-S-transferase (GST) A1. METHODS: CHO cells was transfected with recombinant vector pDNA3.1- GSTA1 by lipofectamineTM 2000. The expression of selected CHO-GSTA1 in G418-resistant clones were determined by PCR, RT-PCR and Western blot. CHO-GSTA1 high-expressed clone was cultured to collect supernatant and activity was assayed by U-2001 ultraviolet spectrophotometer. RESULTS: CHO GSTA1 was obtained by PCR, RT-PCR and Western blot. The expressed protein of GSTA1 had biological activity, it was 35 mmol/(mg min pro). CONCLUSION: The CHO GSTA1 was constructed and for research purification protein and candidate strains of detoxicitic mechanism of chemical toxins and carcinogenesis. PMID- 17424840 TI - [Research on the comet assay in vitro by plating HepG2 cells in 96-well plates]. AB - OBJECTIVE: To provide the proof for the high throughput screening method in vitro by establishing the comet assay in vitro using HepG2 cells plated in 96-well plates. METHODS: Plating HepG2 cells in 96-well plates, and then detecting the cytotoxicity using the neutral red method before comet assay. RESULTS: 8 genotoxicial agents could be detected using the method and the sensitivity was 100% , the ratio of the comet cells and the length of the tail of the comet were increased with the increasing dose when the cytotoxicity less than 50%. 11 non genotoxicial agents could be detected within 12 non-genotoxicial agents and the specificity was 91%. CONCLUSION: (1) This method could detect the direct genotoxic agents and the indirect genotoxic agents at the absent of the S9. (2)4 5 agents could be detected in the same 96-well plate using the method. (3) Using the method, we could obtain an dose at which the agent could cause genotoxicity. (4) By plating cells in plates in vitro, increased the speed of the detection and reduced the consumption of the agents. (5) The method could be developed into the high throughput screening method in vitro. PMID- 17424841 TI - [Study on nitrate-nitrogen removal and community analysis of denitrifying bacteria in downflow fixed-bed bioreactor]. AB - OBJECTIVE: To study the community of denitrifying bacteria and nitrate-nitrogen removal in downflow fixed-bed bioreactor. METHODS: Porous ceramics were used as immobilization carrier. Optimum conditions for nitrate-nitrogen removal of downflow fixed-bed bioreactor were studied. Morphology of immobilized denitrifying bacteria was observed by scanning electron microscope and their community analysis were processed by 16S rDNA sequencing. RESULTS: The optimum conditions for denitrification were T = 35 degrees C and pH8.0. NO3- -N was consumed 99% within the 10h while removal efficiency reached 91% and followed the zero-order kinetics in the first 4h. Majorities of denitrifying bacteria were bacillus. 16SrDNA homology analysis showed that denitrifying bacteria included P. putida, P. nitroreducens, S. maltophilia, C. testosteroni and Ochrobactrum sp etc. CONCLUSION: There were high nitrate-nitrogen removal efficiency and diversity of denitrifying bacteria in downflow fixed-bed bioreactor. PMID- 17424842 TI - [Study on vector construction, expression and refolding of fusion HPT protein]. AB - OBJECTIVE: Hygromycin B Phosphotransferase gene (hpt) is a widely used antibiotic selectable marker gene in the production of genetically engineered crops. Fusion expression vector of the hpt gene was constructed in order to obtain enough HPT protein for further safety assessment of the protein. METHODS: The hpt gene was cloned into liner expression vector pET41 EK/LIC and fusion expression vector pET 41EK/HPT was obtained. The constructed vector was transformed into expression host E. coli BL21 (DE3). After induction, the expressed fusion protein was mainly aggregated as inclusion bodies. To get the active protein, dilution refolding, chromatographic refolding and Sarkosyl resolution and dilution refolding methods were explored. Then the fusion protein was digested by enterokinase and N terminal amino acid sequence was conducted. RESULTS: The refolding study showed the protein got from Sarkosyl resolution and dilution refolding method had higher yield and bioactivity than those from other two methods. From this method 78mg active fusion obtained protein could be obtain from 1L bacterial culture and the purity was about 93%. From N terminal amino acid sequence it was confirmed that after digested by enterokinase, the HPT protein had same N-terminal amino acid sequence as the native HPT. CONCLUSION: The results of this study illustrated that large quantity HPT protein could be acquired through prokaryotic host expression which had comparable molecular weight, N-terminal amino acid sequences and biological activities with those of native HPT protein. PMID- 17424843 TI - [Effect of grape procyanidins on the protein kinase and proliferative cell nuclear antigen protein expression of mice hepatic cells]. AB - OBJECTIVE: To study the effect of grape procyanidins (GPC) on the protein kinase C (PKC) and proliferative cell nuclear antigen (PCNA) expression in mice hepatic cells. METHODS: The mice were orally given at the doses of GPC and killed after 30 days. The proliferation activity of the hepatic cells was determined by MYT assay, and the levels of PKC and PCNA protein expression in the cell were measured by immunohistochemistry method. RESULTS: The expression rates of PKC and PCNA protein of the high-dose GPC group were 47.62% and 82.76% respectively,and those of the negative control group were 6.42% and 32.62%. The differences between the two groups were significant (P < 0.05). The proliferation of mice hepatic cells increased with the dose of GPC in different groups. CONCLUSION: GPC could promote the PKC protein expression and improve the proliferative activity of the mice hepatic cells. PMID- 17424844 TI - [Antagonistic effects of Se-rich Spirulina platensis on rat liver fibrosis]. AB - OBJECTIVE: Antagonistic effects of supplement of Se-rich Spirulina platensis (Se SP) on hepatocirrhosis were investigated with the rat model of liver fibrosis induced by intraperitoneal injection 3% thioacetamide (TAA). METHODS: Parameters of routine liver function, content of malondialdehyde (MDA) and activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) in rat serum were determined by colorimetry. Content of selenium (Se) was measured by DAN fluorometry method and hyaluronic acid (HA) was detected by radio-immunoassay. Liver fibrosis was diagnosed by HE staining and relative contents of collagen (RCC) were estimated by Masson's trichrome staining. RESULTS: Parameters of liver function in Se-SP group were most recovered in all protective groups. Compared with the model groups, contents of MDA and HA were lower, whereas activities of GPx and SOD were higher (P < 0.05) in rats serum of Se-SP group. The RCC in rats liver of Se-SP group were lower than those of the model groups, where the liver fibrosis were identified dominantly to degree I according to pathological diagnosis. Moreover, Se content in rats serum had positive correlation (r = 0.645) with activity of GPx while a negative correlation (r = 0.675) with MDA level. CONCLUSION: The results indicated that Se-SP have detectable antagonistic effects to liver fibrosis, and suggested that enhancement of antioxidation level and liver reserve function might be associated with these effects. PMID- 17424845 TI - [Effect of milk basic protein on rat bone mineral density]. AB - OBJECTIVE: To investigate the effect of milk basic protein (MBP)'s on bone mineral density (BMD) and hemobiochemical indexes in normal and ovariectomized (ovx) rats. METHODS: 48 female Sprague-Dawley rats were ovariectomized and another 12 rats received a sham operation (sham). The ovx rats were randomly separated into 4 groups: ovx control, ovx low-does, ovx media dose, and ovx high dose while 44 normal SD female rats into 4 groups: normal control, normal low dose, normal media-dose, and normal high-dose. MBP at 10, 20 and 30 mg/(kg x d) were applied in rat diet for continuous 90 days. BMD (at proximal, middle of diaphysis, and distal end) and bone mineral content (BMC) was measured by dual energy X-ray absorptiomery. In addition, serum total protein (TP), total cholesterol (CHO), glucose (GLU) were determined. RESULTS: The bone density and mineral composition at distant end of femur in low-dosage BMP group were significantly higher than those in normal control group, while no significant change of bone density was observed with MBP treatment in ovx groups. Total serum protein level and albumin level were significantly higher in MBP treatment groups than those in normal control group. CONCLUSION: MBP at 10 mg/kg bw significantly elevated femoral distant-end bone density, as well as the total blood protein level and albumin level in normal rat. However, similar effect was not observed in ovx rat. PMID- 17424846 TI - [Change of dehydroepiandrosterone in serum of 6 - 18 year-old twin girls in Qingdao City]. AB - OBJECTIVE: To explore the change of serum dehydroepiandrosterone (DHEAS) concentrations throughout the puberty of girls and estimate the effects of genetic and environmental factors on serum DHEAS. METHODS: The study population consists of 360 girls aged 6 to 18 years:132 pairs monozygotic twins and 48 pairs dizygotic twins, who were all from Qingdao city, Shan-dong province. Anthropometric measurement and pubertal development stage were determined by Tanner standard, fasting serum DHEAS contents were assayed by RIA. RESULTS: Serum DHEAS concentrations increased from Tanner I to IV and decreased later, heritability of pubertal girl's serum DHEAS was 0.65. The heritability of 3 different pubertal stages (pre-thelarche, post-thelarche and pre-menarche, and post-menarche) were 0.92, 0.73 and 0.67 respectively. CONCLUSION: Serum DHEAS concentrations of pubertal girls could be mainly influenced by the genetic factors, especially in the period of adrenarche. Therefore, genetic influence might be important in adrenarche-related metabolic disorders. PMID- 17424847 TI - [Changes of cognition and micronutrient levels in rats treated by beta amyloid protein]. AB - OBJECTIVE: To observe the changes of the cognition and micronutrient level in rats treated by beta amyloid protein. METHODS: Thirty male Sprague-Dawley rats were randomly divided into control group, beta amyloid group (Abeta) and saline group. The rats were injected Abeta and saline by bilateral intrahippocampus (10 microg per lateral) in Abeta group and saline group separately. The experiment lasted 2 weeks.The cognition was identified by water maze and step through test. The contents of serum micronutrients were examined by atomic absorption spectrometry and fluorescence spectrometry. RESULTS: Declined cognition and prolonged latency were observed in Abeta group. The contents of serum vitamin C and iron in Abeta group were significantly lower than those of the control group and saline group (P < 0.05). There was no significant difference among the three groups in other indicators. CONCLUSION: It was seemed that impaired cognition and decreases of serum vitamin C and iron level could be observed in rats treated by beta amyloid protein. PMID- 17424848 TI - [Expression and identification of Cry 1Ie in Bacillus coli]. AB - OBJECTIVE: In this study, Cry1Ie expressed protein in the transgenic Cry1Ie maize was used as a model protein to establish a technical platform for the safety assessment of other genetically modified plants. METHODS: Bacillus coli expression system was used to express Cry1Ie protein and then the expressed protein was purified by SDS-PAGE. The equivalence of Cry1Ie proteins expressed in Bacillus coli and in transgenic Cry1Ie maize was analyzed by their immunorecognition and bioactivity. RESULTS: Cry1Ie protein was highly expressed in Bacillus coli and could be segregated and purified by SDS-PAGE. Cry1Ie expressed protein in Bacillus coli is substantially equivalent to that of maize genetically modified with Cry1Ie in tests of immunorecognition western blot and bioactivity in target pest. CONCLUSION: The established technical platform of external Cry1Ie expressed protein can be applied to assess the food safety of genetically modified plants. PMID- 17424849 TI - [Study on inducing differentiation of mouse embryonic stem cells into cardiomyocyte in vitro]. AB - OBJECTIVE: To study the differentiation from embeyonic stem cells into cardiomyocyte. METHODS: Gene alpha/ beta-MHC, a specific gene of cardiomyocyte and another three house-keeping genes were detected by RT-PCR from the ESCs of 0 ,2-,4-,6-,8-, 10-,12-,14-,16-,18- and 20 day and ESCs of 0-,3-, 5- and 10 day with specific culture condition. RESULTS: ESCs started to express alpha/betaMHC gene from 6 day and the levels of alpha/betaMHC increased with culture time. The levels of 14 day's alpha/betaMHC were the highest. After 14 day, alpha/betaMHC gene kept the stable levels. The levels of three house-keeping genes changed significantly with culture time, and GAPDH kept the best stability of three genes. CONCLUSION: The cycle of the differentiation from ESCs into cardiomyocyte in vitro could be designed as 10 - 12 days and GAPDH could be selected as the fittest inner comparison. PMID- 17424850 TI - [Study on the comparison of rDNA and ITS sequence of Fumonisin-producing strains]. AB - OBJECTIVE: To study the phylogenetic relationships of fumonisin-producing Fusarium verticillioides isolates from China and Gibberella fujikuroi species complex. METHODS: 5.8S, 28S rDNA and ITS1 and ITS2 sequences of 4 ATCC typical strains and 12 isolates from different geographical regions and different samples in China and other countries were determined and analyzed. The results were submitted to GenBank. Phylogenetic tree and homology tree were illustrated. The characteristics of etiological distribution of toxigenetic Fusarium verticillioides and the relativity of of rDNA ITS2 sequence and the fumS gene was elucidated by comparing the isolates from China, South Africa and USA. RESULTS: It was found that rDNA highly devergent ITS2 sequences of our isolates with positive fumonisin biosynthase gene were type I, which was consistant with fumonisin-producing isolates from South Africa. The Fusarium verticillioides isolates with ITS2 type II sequence were fumS gene negative. Two Fusarium subglutinans isolates SD-fmv047 and SD-fmv049 were nonfumonisin-producing and ITS2 sequences were neither type I nor II. The DNA sequences had high homology with Fusarium oxysporum. CONCLUSION: New and reliable theory and technology supports were provided for the further study of phylogenetic relationships of Gibberella fujikuroi species complex. PMID- 17424851 TI - [Mimic epitope of aflatoxin B1 screened by phage display technique]. AB - OBJECTIVE: To screen mimic epitope from phage-display random peptide libraries as sources of peptides that mimic the binding of aflatoxin B1 to monoclonal antibodies raised against the toxin and establish immunoassay for aflatoxin B1. METHODS: The monoclonal antibody against the aflatoxin B1 was used as ligand to screen the binding peptide from the Ph.D.-7 peptide library with phage display and the specificity of colons were identified by ELISA. The peptide sequences of positive phage clones were determined and analyzed by DNA sequencing. RESULTS: After four rounds of panning, the binding peptides were screened from random peptide libraries. Through indirect competitive ELISA, 4 positive clones could inhibited aflatoxin B1 and HXXDPXH was characterized by DNA sequencing, X is random amino acid. Then competitive ELISA was established for clone phage 10, the linear ranges of the inhibition curves were between 100 pg/ml and 2000 pg/ml, the detecting limit was 50 pg/ml. CONCLUSION: The mimic epitope of aflatoxin B1 antigen was obtained successfully by the phage display technigue and it is effective substitutes for aflatoxin B1-protein conjugate in the immunoassay. PMID- 17424852 TI - [Antibiotic susceptibility of Enterobacter sakazakii isolated from powdered formula for infant and young children]. AB - OBJECTIVE: To study the antibiotic susceptibility of Enterobacter sakazakii isolated from powdered formula for infant and young children in Chinese market. METHODS: Sixteen strains of E. sakazakii were evaluated for antibiotic susceptibility by disk diffusion technique recommended by the National Committee of Clinical laboratory Standard. RESULTS: All of the isolates were resistant to Benzylpenicillin, Oxaxillin and Vancomycin, moreover cephazolin and Cefpodoxime except one isolate. At the same time, all the isolates were susceptible to at least 13 antibiotics. CONCLUSION: It is important to pay more attention to the use of drugs in prevention and clinic treatment to reduce the multiple-antibiotic resistant strains and failure of experiential treatment. PMID- 17424853 TI - [Study of the expression of exogenous augmenter of liver regeneration recombinant plasmid in liver tissue of rat]. AB - OBJECTIVE: To observe the expression of exogenous ALR recombinant plasmid in liver tissue of rats suffered from immune hepatic fibrosis. METHODS: Immune hepatic fibrosis model of rats were made by using porcine serum, which were treated by pcDNA3-ALR recombinant plasmid. The expression of ALR proteins in liver tissue were detected by immunohistochemistry staining and Western blotting and the expression of ALR mRNA were detected by RT-PCR. RESULTS: ALR were expressed in liver tissue in all group rats. ALR expressions were lower in normal control groups , and were higher in ALR treat groups. The results of image analysis by immunohistochemistry were (0.109 +/- 0.01), (0.159 +/- 0.02) and (0.198 +/- 0.04) in normal groups, model groups and ALR groups respectively. There were significant differences between model and normal groups ( P < 0.01) as well as ALR groups and model groups (P < 0.01). The results of Western blotting were similar to immunohistochemistry. The specific 550bp gene fragments that were amplified by RT-PCR with a couple of especial primers were found in ALR group but not in normal and model groups, which was coincident with fragments amplified by PCR with ALR recombinant plasmid extracted directly from competent E. coli DH5. CONCLUSION: The expression of ALR was relatively low abundance in normal rat liver tissue, and increased markedly in immune injured rat liver tissue. The expression of the pcDNA3-ALR recombinant plasmid can be found in rat liver tissue suffered from immune hepatic fibrosis. The reinforcements of expression of ALR protein in liver tissue of pcDNA3-ALR recombinant plasmid treatment group was resulted from the overlap of endogenous and exogenous ALR expression. PMID- 17424854 TI - [Analyses of anti-hCV detected by ELISA and HCV RNA detected by RT-nPCR in chronic hepatitis C virus infectors]. AB - OBJECTIVE: In order to provide the basis for the clinical test and the blood station screening the health donator, the results of anti-HCV tested by ELISA (enzyme-linked-immuno-absorbed assay ) and HCV RNA tested by RT-nPCR (reverse transcript-nested-polymerase-chain-reaction) were compared in the chronic hepatitis C virus infectors. METHODS: Venous blood samples of 133 chronic hepatitis C virus infectors, 52 health controls were collected in May 2005. These infectors were infected with HCV nearly in 1990 through plasma donator and diagnosed in 1993 in a rural area of Zhao County in Hebei Province, which remained the same diagnosis as HCV infectors in 2002 Hebei Province. The anti-HCV was tested by ELISA and HCV RNA was tested by RT-nPCR. RESULTS: (1) In 185 cases, the positive rates of both anti-HCV and HCV RNA were 49.73% (92/185). The rate of anti-HCV negative but HCV RNA positive was 9.73% (18/185). The rate of anti-HCV positive but HCV RNA negative was 11.89% (22/185). The negative rate of both anti HCV and HCV RNA tested was 28.65% (53/185). The result-agreement rate of ELISA and RT-nPCR methods were 78.38% [(92 + 53)/185]. The disagreement rate between ELISA and RT-nPCR methods was not obviously different (paired chi2 = 0.40, P > 0.05). (2) In the chronic HCV infectors, the sensitivity of anti-HCV tested by ELISA was 82.71%, the specificity was 92.31%, and the omitting rate was 17.29%. The sensitivity of HCV RNA tested by RT-nPCR was 81.20%, the specificity was 96.15%, and the omitting rate was 18.80%. The sensitivity between ELISA and RT nPCR was not obviously different (chi2 = 0.102, P > 0.05). (3) The sensitivity tested by ELISA combined with RT-nPCR was 96.75%, which was evidently higher than that of single ELISA (82.71%) (chi2 = 9.62, P < 0.01). CONCLUSION: The false negative rate was nearly 17% when anti-HCV was tested with single ELISA in HCV infectors. The positive testing rate of HCV infection was increased remarkably when ELISA and RT-nPCR were tested simultaneously. PMID- 17424855 TI - [Study on the status and the relationship between job burnout and work ability in hospital staff in Yinchuan City]. AB - OBJECTIVE: To investigate the status of job burnout and work ability and to explore their relationship in hospitals employee. METHODS: Maslach burnout inventory (MBI) and work ability index (WAI) were used to measure burnout and work ability of 522 doctors and nurse in 10 hospitals in Yinchuan City. RESULTS: The results indicated that, there were remarkable differences between the doctors and nurses in the scores of MBI. The scores of DE and PA of the doctors were remarkable higher than those of nurses (P < 0.01 and P < 0.05). However, in different hospitals, the scores of EE of the doctors in national hospitals were significant higher than those of doctors in private hospitals. But the scores of PA and WAI of the doctors in national hospitals were remarkable lower than those of doctors in private hospitals. In all, the Pearson correlation showed that there are significantly negative correlations between job burnout and work ability (r = - 0.113- - 0.494, P < 0.01). The multianalysis indicated that the variables of influence work ability included the factors of EE, PA, status of sleeping, ages and type of job. CONCLUSION: There are higher job burnout in hospital staff, even so, status of work ability can be improved with enhancing personal resource and achievement, adjusting job burnout and improving the status of sleeping maladjustment. PMID- 17424856 TI - [Preliminary survey on water eutrophication and microcystins level in Beijing Miyun Reservoir]. AB - OBJECTIVE: In order to survey water eutrophication and microcystin levels in Beijing Miyun Reservoir. METHODS: Mivrocystin-LR (MC-LR), and MC-RR were determinated by LC/MS/MS at 5 sample sites in Miyun Reservoir. Eutrophication parameters, which were determinated by conventional analytical methods, included algae number, chlorophyll-a (Chl-a), total phosphor (TP), total nitrogen (TN), COD,TOC, Fe, turbidity and water temperature. RESULTS: Water in Miyun Reservoir was polluted very little by organic matter. According to Chl-a, TP appraise criterion and grading method, Miyun Reservoir was eutrophicated very little. MC LR and MC-RR could be detected out in water of Miyun Reservoir. Maximum concentration was discovered in September, MC-LR 41 ng/L and MC-RR 18 ng/L respectively. CONCLUSION: Beijing Miyun Reservoir was poor-eutrophic. Microcystins concentration in water of Miyun Reservoir was far lower than our country health standard. PMID- 17424857 TI - [Effects of nutrient fortified complementary food supplements on growth of infants and young children in poor rural area in Gansu Province]. AB - OBJECTIVE: To investigate the effects of complementary food supplements with protein and multimicronutrients on growth of infants and yound children in poor rural area in Gansu Province, China. METHODS: In 5 poor counties of Gansu Province, 1478 children aged 4-12 months were enrolled and divided into two groups. In addition to the usual home-made complementary food, all the children were fed one sachet of either formula I or formula II per day, protein and micronutrient were supplemepted in formula I group, fomula II group got energy supplement corresponding with same energy intake as formula I, a massive dose of vitamin A was supplemented to both groups every 6 months interval. Weight and height measurements were done every 3 months until they were 24 months old. RESULTS: At baseline survey, LAZ and WAZ of children in formula II group was better than those of formula I group, the prevalence of under nutrition in formula I was higher than those in formula II group, but not statistically significant. After 12 months supplementation, the differences of LAZ and WAZ between the two groups diminished, the stunting prevalence of the two groups was close, it demonstrated that formula I was slightly more effective than formula II, the effect size was 0.17. Formula I has positive effect independently with the nutrition status of children. When all children were 24 months, formula I was better than formula II in LAZ and WAZ changes P < 0.05), the length changes were bigger in formula I group (P < 0.05). If takeing the children initiating supplementation at under age of 7 month, LAZ changes in formula I group and formula II group was statistically significant (P < 0.005). The average length increased in formula I group was 1.3cm longer. CONCLUSION: Multinutrient fortified complementary food supplement could be effective for children aged 4-12 month in terms of growth promotion. The study proved that application of home level fortification of supplementation into the regular complementary feeding in poor rural China may be feasible. PMID- 17424858 TI - [Trends and prevalence of malnutrition among children under five year-old in Chinese rural of different classes]. AB - OBJECTIVE: To study the trends and prevalence of malnutrition status among Chinese children under 5 year-old in different type rural areas from 1992 - 2002. METHODS: The paper based on anthropometry data collected in 'National Nutrition Survey in 1992' and 'China National Nutrition and Health Survey in 2002' applied age - and sex - specific height and weight reference recommended by WHO/NCHS and analyzed data with the SAS8.2 software. RESULTS: The prevalence of stunting among children under 5 year-old in four classes rural areas were 12.4%, 15.4%, 11.5% and 29.3% respectively, the prevalence of underweight were 7.4%, 8.7%, 5.4% and 14.4%. The prevalence of moderate stunting in Chinese rural areas had decreased from 20.5% in 1992 to 12.0% in 2002, the prevalence of severe stunting fell from 14.5% to 5.2%. Compared to 1992, the prevalence of moderate underweight and severe underweight decreased 47.7% and 72.3% respectively. PMID- 17424859 TI - [Effect of three ways of nutritional education on knowledge-attitude-behavior with nutrition in pupils]. AB - OBJECTIVE: To explore the effects of different nutritional education ways on knowledge-attitude-behavior with nutrition in pupils. METHODS: According to cluster randomization sampling methods, application three ways of nutritional education, nutritional education for pupils, nutritional education for pupils' parents and nutritional education for both pupils and their parents, to 1477 pupils from three, four and five grades in primary school in Xuzhou City. RESULTS: Although each of three ways can improve the level of nutrition knowledge and diet behavior, the way of nutritional education for both pupils and their parents gets the best effect and the following is the way of nutritional education for pupils. CONCLUSION: Primary schools play a more important role than pupils' parents for improving pupils' knowledge, attitude and practice with nutrition. PMID- 17424860 TI - [Comparison between anodic stripping voltammetry and graphite furnace atomic absorption spectrophotometry for blood lead analysis]. AB - OBJECTIVE: To compare and evaluate anodic striping voltammetry (ASV) with graphite furnace atomic absorption spectrophotometry (GFAAS) for blood lead analysis. METHODS: These two methods were used to analyze same samples of three kinds of blood (cattle blood, rat blood & human blood) at the same time. The practicability and reliability of the results were evaluated by the statistic analysis. RESULTS: Limits of detection (LOD) for ASV and GFAAS were 5.3 microg/L and 0.045 microg/L. Limits of quality (LOQ) were 17.8 microg/L and 0.15 microg/L for ASV and GFAAS. The results that three kinds of blood samples were analyzed by these two methods were in high correlation after t test (P > 0.05). CONCLUSION: The data demonstrated that the ASV method could be in the complete conformity to GFAAS for different kinds of blood samples in lead analysis. The results showed that the sensibility of GFAAS was better than that of ASV, but the ASV showed a marked performance in stability, ease of operation and precision as well as GFAAS. GFAAS was suitable for the scientific research as ASV for a clinic. PMID- 17424861 TI - [Study on the di-n-butyl phthalate and di-2-ethylhexyl phthalate level of girl serum related with precocious puberty in Shanghai]. AB - OBJECTIVE: To determine the levels of the di-n-butyl phthalate and di-2 ethylhexyl phthalate in the serum of precocious girls in Shanghai, and study the development relationship between DBP, DEHP and precocious girls. METHODS: The concentrations of two phthalates (di-n-butyl phthalate, DBP, di-2-ethylhexyl phthalate, DEHP) in serum of 110 precocious girls and 100 normal children were measured by using gas chromatography, in the same time, the volumes of the uteruses and ovaries for precocious girls and normal children were measured by B ultrasound and then did correlation analysis between the concentrations of DBP, DEHP and the volumes of uteruses and ovaries of precocious girls. RESULTS: DBP and DEHP were detected for 27.3% and 22.7% in precocious girls respectively, DBP and DEHP were detected for only 4% and 3% in normal children respectively. From the ratio of DBP and DEHP in serum of precocious girls were higher than normal children. Precocious girls uteruses and ovaries volume were higher than those of normal children respectively (P < 0.05, P < 0.05). DBP in serum of precocious girls had positive correlation with the volumes of the uteruses (r = 0.456, P < 0.05), and had positive correlation with volumes of the ovaries (r = 0.378, P < 0.01). DEHP in serum of precocious puberty had positive correlation with the volumes of uteruses (r = 0.382, P < 0.05), and had positive correlation with volumes of ovaries (r = 0.689, P < 0.01). CONCLUSION: Precocious girls could be contaminated more severely than normal girls, and DBP, DEHP could influence the volumes of the uteruses and ovaries. PMID- 17424862 TI - [Research progress on epigenetics of lung cancer]. AB - Epigenetic events, a key driving force in the development of lung cancer, two changes integral to epigenetic transcriptional control are DNA methylation and covalent modification of histone proteins. Aberrant methylation may be the most common mechanism of inactivating cancer-related genes in lung cancer, and histone modification may be closely associated with DNA methylation. It was seemed that epigenetic changes could be of the earliest events observed during cancer development, making them excellent targets for chemoprevention. Understanding the mechanisms involved in epigenetic regulation and how they interact with genetic changes during lung cancer progression could facilitate development of newer, more efficacious, and safer chemopreventive agents. PMID- 17424863 TI - [Relation between cytochrome P450 2D6 and lung cancer susceptibility caused by smoking]. AB - Cytochrome P450 2D6(CYP2D6) is one of the prominent phase I metabolizing enzymes in hepatic tissue, and is responsible for metabolically activating the tobacco specific nitrosamine such as NNK and nicotinamide to highly reactive metabolites, so it may play a role in lung cancer. The study of its biological characters, structure function and the relationship beween CYP2D6 genetic polymorphisms and lung cancer susceptibility can redound to understand the significant influence of CYP2D6 on lung cancer susceptibility. Furthermore from the point of view of interaction between gene and environment it can evaluate the lung cancer risk accurately. PMID- 17424864 TI - [Advance of study on the detoxification of the microcystin]. AB - Contamination of water on toxic blooms of cyanobacteria has occurred in many regions of the world and could be a serious public health problem. It is hard to degrade all the microcystins by using ordinary waste water treatment process, so it is increasingly urgent to develop a new technology to deal with them. In this article the current ways of effectively removing microcystin toxins were reviewed. PMID- 17424865 TI - [Advances on the level of reference nutrient intake of protein and amino acid]. AB - The value of requirement of protein in adult is 0.66g/(kg x d), as is suggested by FAO in 1957. Through the detail discusses repeated by many experts in the world, FAO/WHO/UNU suggested that RNI of protein and amino acid was different for different age groups, based on nitrogen balance method and factorial method. But there were opposited opinions, based on the same methods, the values that some research groups in America and England got were different from that of FAO/WHO/UNU. The requirements of specific indispensable amino acid that Vernon. R. Young had got by using the stable isotope technique were 2 - 3 times higher than that of FAO/WHO/UNU. PMID- 17424866 TI - [Relationship between ghrelin and food intake or obesity]. AB - Ghrelin, an endogenous ligand of the growth hormone secretagogue receptor, is mainly produced by the stomach, but its expression has also been demonstrated in many other tissues. It has diverse biological effects. ghrelin stimulates food intake and regulates energy homeostasis through activating the expression of the orexigenic neuropeptides NPY (neuropeptide Y)/AGRP (agouti-related protein) in hypothalamic neurons, by which it plays a key role in obesity pathogenesis. This article simply introduces the recent research on its molecular structure, distribution, physiological effects and highlights the relationship between ghrelin and food intake as well as obesity. PMID- 17424867 TI - Nasal contact endoscopy for the in vivo diagnosis of inverted schneiderian papilloma and unilateral inflammatory nasal polyps. AB - BACKGROUND: Inverted schneiderian papilloma is an entity surrounded by controversies ranging from its etiology to the indication of treatment. Any method that permits histopathological analysis without damage to the tissue studied deserves to be investigated. In this study, we tested a new noninvasive method, i.e., nasal contact endoscopy. METHODS: The main alterations visible on nasal contact endoscopy were described in 11 patients with inverted papilloma and in 8 patients with unilateral inflammatory nasal polyps. The characteristics showing the differentiation between the two entities were defined and these findings were presented to inexperienced examiners. RESULTS: The significant characteristics for differentiation between the two diseases were the presence of vacuolized cells, cellular heterogeneity, cell clusters, a ciliated respiratory epithelium, and vascular patterns. Two of the three examiners showed a statistically significant high rate of correct diagnosis. CONCLUSION: Nasal contact endoscopy may be an effective method for the differential diagnosis between inverted schneiderian papilloma and inflammatory polyps even when the examiner is inexperienced. PMID- 17424868 TI - Activity of nasal amphotericin B irrigation against fungal organisms in vitro. AB - BACKGROUND: The role of fungi in chronic rhinosinusitis has been described in recent reports. Controversy exists on the use of topical amphotericin B therapy as a treatment modality for this condition. The effect of various concentrations of amphotericin B nasal irrigation on actively growing fungi was studied in vitro. METHODS: Ten species of fungi commonly found in the nasal cavity were grown on growth media plates. Each fungi was exposed to 20 mL of amphotericin B nasal irrigation at concentrations of either 100, 200, or 300 microg/mL or sterile water two times daily for 6 weeks. Each plate was subcultured on a weekly basis to examine for any viable fungi. RESULTS: Fungi growth was not arrested in the 100-microg/mL amphotericin B and sterile water groups at the end of 6 weeks. Use of the 300-and 200-microg/mL amphotericin B solutions showed failure of the subcultured fungi to grow at 5 and 6 weeks, respectively. CONCLUSION: Nasal amphotericin B irrigation is ineffective in killing fungi in vitro at a concentration of 100 microg/mL over a 6-week period. Concentrations of 200 and 300 lig/mL successfully prevented fungi growth at the conclusion of the study. The current concentration of commercially available topical amphotericin B (100 microg/mL) seems ineffective in eradicating fungi in vitro. PMID- 17424869 TI - Effectiveness of topical antibiotics on Staphylococcus aureus biofilm in vitro. AB - BACKGROUND: In vitro biofilm-producing capacity in isolates of Staphylococcus aureus and Pseudomonas aeruginosa collected from the sinus cavities after endoscopic sinus surgery (ESS) are associated with a poor outcome in patients with chronic rhinosinusitis (CRS). However, conventional oral antibiotic therapy is frequently ineffective in eradicating bacteria in the biofilm form. Increasing the concentration of antibiotics may offer a means of countering this resistance. The aim of this study was to determine the in vitro activity of moxifloxacin (MOXI) against S. aureus in biofilm form (recovered from patients with CRS at least 1 year post-ESS). METHOD: This study was performed in a research microbiology laboratory, where five isolates of S. aureus with known biofilm forming capacity were cultured in Tryptic Soy Broth 0.5% glucose in 96-well plates at 37 degrees C for 24 hours. After visual confirmation of biofilm formation, plates were incubated in phosphate-buffered saline (PBS) or with MOXI at concentrations of 0.1x, Ix, 100x, and 1000x minimal inhibitory concentration (MIC) for an additional 24 hours. Biofilm from 3 wells of each concentration were collected and sonicated and the number of viable bacteria was determined by serial dilution and plating. RESULTS: After incubation, the number of viable bacteria was similar for nontreated and MOXI-treated biofilms at MIC and sub-MIC levels. However, MOXI at 1000x (0.1-0.2 mg/mL) gave a 2 to 2.5 log reduction in number of viable bacteria. CONCLUSION: In vitro results show that increased concentrations of antibiotics, easily attainable in topical solutions, are effective in killing bacteria in bacterial biofilms. This suggests a role for topical antibiotic therapies in the treatment of biofilm infections. PMID- 17424870 TI - Outpatient intravenous antibiotics for methicillin-resistant Staphylococcus aureus sinusitis. AB - BACKGROUND: The widespread use of broad-spectrum antibiotics has resulted in an increase in the prevalence of methicillin-resistant Staphylococcus aureus (MRSA). Its presence in sinonasal cultures in patients with sinusitis suggests its pathogenicity. However, the efficacy and safety of treatment modalities for MRSA sinusitis remain incompletely described. METHODS: A retrospective chart review of six patients treated for MRSA sinusitis with outpatient intravenous (i.v.) antibiotics was performed for patient demographics, history of antibiotic use, history of prior sinus surgery, and treatment-related complications. A quality-of life survey and endoscopically guided cultures before and after therapy were used to measure treatment outcomes. RESULTS: The cohort consisted of five women and one man with a mean age of 50.8 years. All patients had undergone multiple sinus procedures with a mean number of 2.7 procedures per patient (range, 1-6 procedures). Five patients (83.3%) experienced negative cultures after outpatient i.v. antibiotics. The single patient who had persistent cultures experienced clinical and endoscopic improvement in her symptoms. The quality-of-life scores improved in five of the six patients (83.3%) after therapy. Four patients (66.7%) experienced five adverse events including allergic reaction (four events) and neutropenia (one event), all of which resolved with a change in medication. CONCLUSION: Outpatient i.v. antibiotics may be an effective therapy for the treatment of MRSA sinusitis. The occurrence of adverse events requires a dedicated protocol to therapy. Future studies are required to investigate long term efficacy. PMID- 17424871 TI - Clinical predictors of chronic rhinosinusitis. AB - BACKGROUND: Chronic rhinosinusitis (CRS) is diagnosed by the presence of signs and symptoms of CRS in conjunction with physical evidence of mucosal inflammation. We sought to identify symptoms that predict CRS. METHODS: We performed a retrospective review of patients referred to a tertiary care rhinology clinic for evaluation of CRS. Symptom survey data and diagnoses were reviewed. RESULTS: Of 187 patients who met inclusion criteria, 112 (60%) were diagnosed with CRS and 75 (40%) were not. Chronic purulent rhinorrhea (odds ratio [OR], 2.2) and hyposmia (OR, 2.3) individually and in combination (OR, 3.8) were significant predictors of CRS. The major criteria of the 1997 Task Force in Rhinology (TFR) also predicted CRS (OR, 1.9) but the minor criteria did not (OR, 0.3). CONCLUSION: In this preliminary study, purulent rhinorrhea and hyposmia are strong predictors of CRS. The major criteria of the TFR also predict CRS but the minor criteria do not. PMID- 17424872 TI - Choanal polyps: an evaluation of 53 cases. AB - BACKGROUND: The aim of this study was to evaluate our experience on the diagnosis and treatment of choanal polyps (CP). METHODS: This study consists of 53 patients with CP diagnosed by means of rhinoscopy, nasal endoscopy, computerized tomography, surgical, and histologic findings between 1996 and 2005. RESULTS: The origins of the CPs were the unilateral maxillary sinus in 47 patients, simultaneously bilateral maxillary sinus in 2 patients, septum in 2 patients, sphenoid sinus in I patient, anterior ethmoid sinus in 1 patient. The most common presenting symptoms were unilateral nasal obstruction, snoring, and rhinorrhea. In the cases of the antrochoanal polyps, the most common preoperative radiological finding was the total opacification of the maxillary sinus. CONCLUSION: Endoscopic approach for complete removal of the CPs is an extremely safe and effective procedure. It should be focused on the detection of the exact origin and the extent of the polyp to prevent recurrence. PMID- 17424873 TI - Smoking and improvement after nasal surgery in snoring men. AB - BACKGROUND: The aim of this study was to compare subgroups of smokers and nonsmokers undergoing nasal surgery and to evaluate improvement of nasal stuffiness, snoring, and symptoms related to sleep-disordered breathing after nasal surgery. METHODS: A cross-sectional prospective study was performed. The study population included 40 consecutive snoring men scheduled for surgical treatment of nasal obstruction. The patients completed nasal and sleep questionnaires, an Epworth sleepiness scale, and a visual analog scale of snoring intensity. They underwent polysomnography, anterior rhinomanometry, acoustic rhinometry, and cephalometric analysis. RESULTS: The smokers were younger, they snored longer and louder, and they had higher nasal resistance with decongestion and longer soft palates than the nonsmokers. Nasal stuffiness improved well after surgery, but a decrease of nasal resistance was not related to improvement of subjective snoring. CONCLUSION: Smoking was associated with increased snoring, nasal obstruction, and pharyngeal soft tissue volume. Expectations of patients may influence subjective assessment of snoring after nasal surgery. PMID- 17424874 TI - Synechia formation after endoscopic sinus surgery and middle turbinate medialization with and without FloSeal. AB - BACKGROUND: The aim of this study was to determine the incidence, outcomes, and risk factors for synechia formation after endoscopic sinus surgery (ESS) and middle turbinate medialization with and without FloSeal. METHODS: A retrospective review was performed of patients who underwent primary ESS with middle turbinate medialization, with or without the placement of FloSeal. Medialization was performed with the placement of an absorbable conchopexy suture and silastic splint. Operative variables and outcomes were analyzed to identify risk factors for synechia formation. RESULTS: One hundred thirty-five patients underwent medialization alone and 37 patients underwent medialization with placement of FloSeal. Overall, synechia formation was noted in 16 patients (9.3%). A statistically significant higher incidence of synechia formation was noted in patients who underwent middle turbinate medialization with the placement of FloSeal versus medialization alone (18.9% versus 6.7%). The incidences of intraoperative complications (6.2% versus 4.7%) and postoperative complications (6.2% versus 7%) were similar between patients with and without synechia, respectively. Patients experiencing synechia, however, underwent a statistically significant higher rate of revision procedures (25% versus 5.1%). CONCLUSION: Despite adequate prevention with middle turbinate medialization, synechia formation after ESS may result in higher rates of revision procedures. The placement of FloSeal in conjunction with middle turbinate medialization may result in a higher incidence of synechia formation. PMID- 17424875 TI - Evaluation of the effects of nasal septal deviation and its surgery on nasal mucociliary clearance in both nasal cavities. AB - BACKGROUND: In this study the effects of nasal septal deviation (NSD) and its surgery on nasal mucociliary clearance velocity (NMCV) in both nasal cavities were evaluated. METHODS: This study included 20 patients with NSD and 20 healthy subjects as a control group who had no complaint related to the nose. NMCV was measured with rhinoscintigraphy using technetium-99m-macroaggregated albumin (Tc 99m-MAA). RESULTS: Before the septal surgery, the NMCV of concave (10.24 +/- 3.96 mm/minute) and convex sides (10.78 +/- 3.53 mm/minute) of the patients were significantly lower than the control group (17.94 +/- 2.89 mm/minute). There is no statistically significant difference between the NMCV of the concave and convex sides. After septal surgery, the NMCVs of the concave (16.34 _ 4.40 mm/minute) and convex sides (17.21 +/- 3.43 mm/minute) were not significantly different from control groups (17.94 +/- 2.89 mm/minute). Postoperative NMCVs of the concave and convex sides was significantly better than preoperative NMCVs. CONCLUSION: The results of this study showed that NSD significantly impaired NMCVs in both sides and that septoplasty significantly improved NMCVs in both sides. PMID- 17424876 TI - Use of a hypodense sodium fluorescein solution for the endoscopic repair of rhinogenic cerebrospinal fluid fistulae. AB - BACKGROUND: Hypodense fluorescein solution can be used intrathecally to facilitate and accelerate the identification of the fistulous site. METHODS: Eighteen patients were submitted for nasal endoscopic correction of rhinogenic cerebrospinal fluid fistulas after their identification with a hypodense sodium fluorescein solution. RESULTS: Intrathecal injection of hypodense fluorescein permitted a rapid and safe identification of the leak and did not present significant side effects. The fistulous site was identified in all patients, and the time needed for staining was <30 minutes in all cases. Surgery was successful in 88% of the patients after only one intervention, with the rate reaching 100% after a secondary intervention in cases of recurrence. CONCLUSION: Intrathecal injection of a hypodense sodium fluorescein solution permits a more precise, rapid, and safe endoscopic approach, with no need to place the patient in the Trendelenburg position or to wait for a long period of time before starting the surgical procedure. PMID- 17424877 TI - Use of a sprayed fibrin hemostatic sealant after laser therapy for hereditary hemorrhagic telangiectasia epistaxis. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) is a relatively common autosomal dominant condition. Epistaxis is a frequent manifestation, often occurring daily and requiring iron and blood transfusions. Surgery often is bloody and difficult. The aim of this study was to evaluate the effectiveness of a sprayed fibrin, hemostatic sealant in preventing postoperative epistaxis after laser treatment of nasal mucosa in HHT. Fibrin sealant was compared with nasal packing for likelihood of postoperative epistaxis and financial impact including material costs and hospitalization fees. METHODS: Retrospective review was performed of 64 individual laser treatments for epistaxis in HHT patients at the University of California, San Diego, Medical Center between 2002 and 2005. Nasal packing was used in 30 procedures and fibrin sealant was used in 34 procedures. RESULTS: Six of 30 (20%) procedures using postoperative nasal packing required admission with an average hospital expense of $5914. One of 34 patients (3%) in the fibrin sealant group required hospitalization (p = 0.04). CONCLUSION: Aerosolized fibrin sealant prevents postoperative epistaxis after nasal laser treatment in HHT patients. Compared with traditional nasal packing we found improved patient comfort and recovery with substantial cost savings. PMID- 17424878 TI - Reduced nasal resistance after simple tonsillectomy in patients with obstructive sleep apnea. AB - BACKGROUND: The aim of this study was to investigate the effects of simple tonsillectomy on nasal resistance in patients with obstructive sleep apnea syndrome (OSAS). METHODS: Conventional tonsillectomy was performed in 20 patients who were refractory to treatment with continuous positive airway pressure. The subjects consisted of 17 men and 3 women (mean age, 32.9 +/-6.3 years). The effects of tonsillectomy were evaluated with preoperative and postoperative polysomnography and nasal resistance. RESULTS: After tonsillectomy, nasal resistance decreased significantly from 0.39+/-0.30 Pa/cm(3) per second to 0.27 +/-0.16 Pa/cm(3) per second (p < 0.05). Simultaneously, the apnea-hypopnea index decreased significantly from 55.7 +/-22.5 to 21.2 +/-14.2 (p < 0.05). There was no significant correlation between tonsillar weight and percentage of change in bilateral nasal resistance (p > 0.05). CONCLUSION: The reduction in nasal resistance induced by simple tonsillectomy could play an important role in improving OSAS, as does nasal surgery or adenotomy. PMID- 17424879 TI - Deposition of aerosolized particles in the maxillary sinuses before and after endoscopic sinus surgery. AB - BACKGROUND: Recent studies suggest that topical therapy is beneficial in many conditions underlying chronic sinusitis. Current literature has documented low aerosolized particle deposition efficiency into the paranasal sinuses. Mathematical modeling suggests that three factors influence the deposition efficiency: particle size, pressure gradient, and size of the sinus ostium. Ostium size is the most dominant factor. Therefore, we sought to determine if maxillary antrostomy and ethmoidectomy would increase the deposition efficiency. METHODS: Five cadavers underwent pre- and postoperative scintigraphy after administration of aerosolized Tc-99M. Images were obtained with a gamma-camera and regions of interest (ROIs) were drawn around the maxillary sinuses. Counts per minute in the pre- and postoperative ROIs were then compared using the paired t-test. RESULTS: Results indicated a significant increase in deposition of radioactivity in the maxillary sinuses in the postoperative state (p < 0.01). CONCLUSION: Topical therapy for chronic sinusitis may be more feasible in the postoperative population. PMID- 17424880 TI - Patient satisfaction survey of outpatient-based topical local anesthetic KTP laser inferior turbinectomy: a prospective study. AB - BACKGROUND: The aim of this study was to determine patient satisfaction and feasibility of performing KTP laser inferior turbinectomy under topical anesthetic/decongestant spray. A prospective survey study was performed in the Outpatients' Department of a District General Hospital. METHODS: Thirty-nine consecutive patients suffering from allergic or nonallergic vasomotor rhinitis were prospectively entered into the study. Preoperative details were gathered including current nasal obstruction score as indicated on a visual analog score (VAS). Lidocaine hydrochloride 5% w/v and phenylephrine hydrochloride 0.5% w/v was applied via topical spray to anesthetize and decongest the nose. The inferior turbinates (ITs) were reduced by parallel mucosal cuts using the KTP laser. Intraoperative details were recorded included pain experienced and bleeding. We studied the subjective clinical effectiveness and patient satisfaction. RESULTS: All patients were followed up at standard intervals by telephone over a 6-month period. At each interval, current nasal obstruction, pain, bleeding, and crusting on VAS were collected. Morbidity was determined in terms of return to work, return to hospital, or recurrence of symptoms. At the end of 6 months all patients were asked if they would have the procedure repeated again and/or recommend it to a friend. Reduction of ITs under topical anesthesia was acceptable to 83% of all patients. Eighty-seven percent of all patients felt they would recommend the procedure and mode of anesthesia to a friend. Our patients obtained the maximum subjective benefit at 6 weeks postoperatively. A significant improvement in nasal obstruction was achieved. There was subjective improvement in associated symptoms including hyposmia/anosmia, rhinorrhea, and sneezing. Only 28% of all patients required medication again to aid symptom control. Postoperative bleeding was insignificant. The return to a ward or clinic was minimal and 82% of all employed patients were back at work within a week. CONCLUSION: Our study showed that KTP laser inferior turbinectomy is suitable to be performed in the outpatient department under topical anesthetic/decongestant spray and provides excellent relief for nasal obstruction and high patient compliance and satisfaction. PMID- 17424881 TI - Comparison of techniques for transsphenoidal pituitary surgery. AB - BACKGROUND: The aim of this study was to compare three different techniques for transsphenoidal pituitary surgery: (1) sublabial transseptal approach with microscopic resection, (2) transnasal transseptal approach with endoscopic resection, and (3) endoscopic approach with endoscopic resection. METHODS: We performed a retrospective review of 50 pituitary surgeries performed by the same neurosurgeon. Demographic, radiographic, and clinical data were collected. RESULTS: Fifteen patients underwent sublabial approach with microscopic tumor resection, 21 patients underwent the transnasal approach with endoscopic resection, and 14 patients underwent the completely endoscopic technique. There were a total of 20 complications in the sublabial group, 13 transnasal complications, and 6 endoscopic complications. Cerebrospinal fluid leak incidence was 53% in the sublabial approaches, 47% transnasal, and 28% in the endoscopic patients. Diabetes insipidus was encountered in 33% of sublabial approaches, 5% of transnasal approaches, and 7% of endoscopic approaches. Lumbar drains were required in 40% of sublabial approaches, 38% of transnasal approaches, and 7% of endoscopic approaches. Nasal packing was used in 100% of sublabial and transnasal approaches and 0% of endoscopic approaches. Mean recurrence rate and follow-up was sublabial in 6.6% (50 months), transnasal in 9.5% (11 months), and endoscopic in 0% (7 months). Average hospital stay for sublabial approaches, transnasal approaches, and endoscopic approaches was 8.3, 6.2, and 3.4 days, respectively (p < 0.05). CONCLUSION: Transsphenoidal pituitary surgery has evolved over the past several decades, because advances in technology have been the catalyst for minimally invasive surgeries. Less invasive approaches, such as the transnasal approach with endoscopic resection of tumor and the completely endoscopic .technique have less morbidity and a shorter hospital stay than traditional sublabial approaches. Continued follow-up is needed to confirm long-term benefits and similar recurrence rates. PMID- 17424882 TI - Impact of age on presentation of chronic rhinosinusitis and outcomes of endoscopic sinus surgery. AB - BACKGROUND: As the population ages, the mean age of patients with chronic rhinosinusitis (CRS) likely will increase as will the frequency of endoscopic sinus surgery (ESS) in the older population. The purpose of this study was to compare symptom presentation, CT scores, endoscopy scores, and quality of life (QOL) measures in patients >60 years of age as compared with younger patients. METHODS: A prospective cohort of 18 patients >60 years and 121 patients < or =60 years undergoing ESS for CRS were studied. Patient factors, symptom severity, and CT scores were examined preoperatively. Endoscopy and QOL scores were compared pre- and postoperatively between the two groups. RESULTS: The prevalence of allergies, polyps, asthma, aspirin sensitivity, and revision surgery was similar in each age group. There was no statistically significant difference in the visual analog scale scores of symptom severity in each age group. Older patients had scores that were similar to younger patients with regard to CT (13.11 versus 11.63; p = 0.419), preoperative endoscopy (8.22 versus 7.96; p = 0.828), postoperative endoscopy (3.06 versus 4.82; p = 0.100), and change in endoscopy ( 5.17 versus -3.15; p = 0.083). Older patients had similar preoperative, postoperative, and change scores when compared with younger patients on the Rhinosinusitis Disability Index (-15.33 versus -20.63; p = 0.318) and Chronic Sinusitis Survey (14.35 versus 23.11; p = 0.126). CONCLUSION: Older patients with CRS present with symptoms and patient factors that are comparable with younger patients and have a similar degree of improvement on endoscopy scores and QOL measures after ESS. PMID- 17424883 TI - Randomized, controlled, study of absorbable nasal packing on outcomes of surgical treatment of rhinosinusitis with polyposis. AB - BACKGROUND: The aim of this study was to establish the effect of absorbable dressing on postoperative discomfort and mucosal healing after sinus surgery. A prospective, randomized, controlled, blinded study was performed. METHODS: Patients 18-80 years old undergoing sinus surgery were enrolled in the study. Each patient's ethmoid cavities were randomized to receive either absorbable dressing or the standard nonabsorbable sinus packs. Therefore, patients served as their own control. Preoperative as a CT scan and intraoperative endoscopic photographs were used for staging within the Lund-Mackay system. The procedure was performed as the indicated by extent of disease. The remaining absorbable dressing was removed at 2 weeks by endoscopic suctioning in the clinic. Patients completed questionnaires regarding sinus symptoms and discomfort. Postoperative endoscopic appearance was graded by a single rhinologist. Length of follow-up was 6 months. RESULTS: Thirty-five patients were randomized. There were no significant adverse events in either group. Patients' symptom scores improved at 2 weeks and at 1 and 3 months when compared with preoperation. Both groups had similar preoperative grade of disease and extent of surgery. Endoscopic appearance of the absorbable cavity showed a trend toward improvement at 2 weeks (p < 0.05). Endoscopic appearance showed a similar trend toward improvement at 1, 3, and 6 months in the absorbable group (NS). Twenty-seven patients had a strong preference for a particular nasal packing of which 16 of 27 (59.3%) patients preferred the absorbable dressing. CONCLUSION: The absorbable dressing showed a trend toward positive effect on early wound healing and in late results. Strong patient preference was indicated for the absorbable dressing over standard sponges. PMID- 17424884 TI - Cigarette smoke extract induces cytotoxicity on human nasal epithelial cells. AB - BACKGROUND: The nasal mucosa is the first area to be exposed to a variety of inhaled toxins. Among various inhaled toxins, cigarette smoke is the most common one and is associated with several nasal and sinus disorders. METHODS: To evaluate the cytotoxic effects of cigarette smoke, primary human nasal epithelial cells were cultured in various concentrations of cigarette smoke extract (CSE) for various times. Cell viability was evaluated by WST-1 (4-[3-(4-iodophenyl)-2 (4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) assay. Morphological findings were observed under the light microscope and the electron microscope. Annexin-V stain was used for the detection of apoptosis. RESULTS: Using the WST-1 assay, CSE reduced cell viability in a time- and concentration-dependant manner. CSE-treated cells showed initial membrane blebbing followed by vesicle formation without apoptotic body formation or cell membrane rupture. Cells were stained with annexin-V but without propidium iodide under a fluorescence microscope. TUNEL (terminal deoxynucleotidyltransferase-mediated dUPT nick end labeling) stain was positive in CSE-treated cells. CONCLUSION: CSE induces cytotoxicity on primary human nasal epithelial cells and the morphological findings closely mimic partition apoptosis. PMID- 17424885 TI - Innate responses to Aspergillus: role of C1q and pentraxin 3 in nasal polyposis. AB - BACKGROUND: Pentraxin 3 (PTX3) and complement component C1q are humoral factors of innate immunity, produced at sites of inflammation, and are essential in immune defense against several microbes such as Aspergillus, which is commonly implicated in nasal polyposis. METHODS: PTX3 and C1q were measured in nasal polyp tissue, normal nasal mucosa, and serum of patients and healthy subjects. Immunohistochemistry for the two proteins was done on normal nasal mucosa and nasal polyps. In addition, PTX3 and C1q production from mononuclear cells from patients and healthy subjects was assessed. RESULTS: Normal nasal mucosa was found to have 100-fold higher levels of PTX3 compared with serum. No measurable local increase of PTX3 was observed in polyps compared with normal mucosa. Immunohistochemistry revealed PTX3 expression in the lining of blood vessels both within normal mucosa and nasal polyps. PTX3 also was present in mononuclear cells infiltrating nasal polyps. C1q levels were higher in polyps than in normal nasal mucosa. CONCLUSION: High levels of PTX3 are present in normal nasal mucosa, suggesting a role in the maintenance of tissue homeostasis. Elevated C1q levels in nasal polyps might be indicative of an ongoing inflammatory response in the nasal mucosa in these patients. PMID- 17424886 TI - Microarray cDNA to identify inflammatory genes in nasal polyposis. AB - BACKGROUND: The objective of this study was to investigate the spectrum of inflammatory gene expression in patients with nasal polyposis. METHODS: The cDNA microarray technique was used to identify gene expression in tissue samples from nasal polyps and adjacent inflammatory nasal mucosa of 21 patients with nonallergic nasal polyposis. To validate the microarray analysis, we compared the expression of five genes by reverse transcription-polymerase chain reaction (RT PCR): tumor necrosis factor, IL-5, IL-9, fibroblast growth factor, and transforming growth factor (TGF)-beta1. RESULTS: We tested 96 different inflammatory genes in our samples. Thirty-six genes exhibited differences in expression between the two tissue types. In all 36 genes the level of expression was greater in the inflammatory mucosa than the polyps. The RT-PCR confirmed the cDNA results. CONCLUSION: We believe that the high expression of TGF-beta1 in inflammatory mucosa compared with the low expression in polyps may reflect an important role in the inhibitory mechanisms of nasal polyposis. PMID- 17424887 TI - Assessment for microsatellite DNA instability in nasal cytology samples of patients with allergic rhinitis. AB - BACKGROUND: Genetic alterations, including microsatellite instability (MSI) and loss of heterozygosity (LOH), have been described in both malignant and benign diseases. Previous studies have successfully detected such alterations in sputum samples of patients with bronchial asthma (BA). The aim of this study was to assess the presence of MSI and/or LOH in nasal cytology samples of patients with allergic rhinitis (AR). METHODS: Nasal brush samples and peripheral blood from 20 patients with AR were analyzed. DNA was extracted and analyzed for MSI and LOH using the following microsatellite markers: D16S289, D4S2394, D4S1651, DXS8039, D3S3606, and D2S2113, harboring potential susceptibility genes for AR and atopy. Microsatellite analysis was performed also in eight control subjects. RESULTS: No MSI and/or LOH were noted in either the AR or the control group. CONCLUSION: Although MSI and LOH are detectable phenomena in sputum samples of patients with BA, this seems not to be the case for nasal cytology samples of patients with AR. Additional studies are needed, using a larger number of polymorphic markers, to assess if such a difference exists among two diseases otherwise very closely related. PMID- 17424888 TI - STAT1 is involved in the pathogenesis of murine allergic rhinitis. AB - BACKGROUND: Signal transducer and activator of transcription (STAT) 1 signaling pathway mediates biological functions of interferon (IFN) gamma, which is a key cytokine-regulating T helper 1 (Thl) differentiation. Although constitutive activation of STAT1 has been reported in the airway epithelium of patients with chronic asthma, its in vivo role in the pathogenesis of allergic rhinitis is not clear. We determined the role of STAT1 in the pathogenesis of allergic rhinitis in vivo using STAT1 gene-deficient (STAT1-/-) mice and a murine model of Schistosoma mansoni egg antigen (SEA)-induced allergic rhinitis. METHODS: STATI /- BALB/c and wild-type (WT) mice were sensitized by intranasal administration of SEA, and their immunologic responses were examined. RESULTS: STATI-1- mice showed impaired nasal eosinophilia and markedly reduced histamine-induced nasal hyperresponsiveness after SEA sensitization. Moreover, levels of Th2-associated SEA-specific IgG1 and IgE antibodies were lower in STAT1-/- mice. Anti CD3stimulated nasal lymphocytes from STAT1-/-mice also produced less amounts of Th2-associated cytokines IL-4, IL-5, IL-10, and IL-13 compared with WT mice, but both produced comparable levels of IFN-gamma. CONCLUSION: These results show that STAT1 is involved in the pathogenesis of SEA-induced allergic rhinitis in BALB/c mice. Furthermore, they reveal a surprising role of STAT1 in induction of nasal eosinophilia, and Th2-type cytokine production from nasal lymphocytes during allergic rhinitis. PMID- 17424889 TI - Relationship between Toxocara seropositivity and allergic rhinitis. AB - BACKGROUND: Allergic rhinitis is the most common chronic disease in the upper respiratory tract, requiring better diagnostic methodology to counteract its onslaught. The purpose of this study was to investigate the association between Toxocara seropositivity and allergic rhinitis. METHODS: Sixty-four consecutive patients with allergic rhinitis and 61 healthy volunteers in the control group were included in this study. The diagnosis of allergic rhinitis was defined by positive epicutaneous prick test. The diagnosis of toxocariasis was established by IgG and specific IgE antibodies against Toxocara by ELISA method. All cases were examined for intestinal parasites also. The seropositive cases with Toxocara were reevaluated for symptoms and signs of toxocariasis. RESULTS: Higher toxocariasis seropositivity (ELISA IgG; 28.1%) rate was determined in the patients with allergic rhinitis and compared with the control groups (11.5%; p < 0.05). Specific IgE seropositivity (17.2%) was high for the patients with allergic rhinitis compared with the control group (3.3%; p < 0.05). CONCLUSION: Our results show that there was a possible causal relationship between Toxocara infection and allergic rhinitis. Patients with allergic rhinitis from a region with high prevalence for this parasitic infection should be evaluated for Toxocara infection. PMID- 17424890 TI - Tumor necrosis factor-alpha stimulates the expression of C-C chemokine ligand 2 gene in fibroblasts from the human nasal polyp through the pathways of mitogen activated protein kinase. AB - BACKGROUND: Recruitment of macrophages is crucial to the pathogenesis of the nasal polyp (NP) because this disease is believed to be inflammation related. Information regarding the expression of C-C chemokine ligand 2 (CCL2), an essential modulator of monocyte chemotaxis in nasal polyp fibroblasts (NPFs), remains unavailable. In this study, the effects of tumor necrosis factor (TNF)-a on CCL2 expression in NPFs and the signaling pathway involved were investigated. METHODS: Primary cultures of NPFs were established from NPs. The expressions of CCL2, c-Fos, and c-Jun mRNAs in NPF after TNF-a stimulation were detected by Northern blot. Western blot was used to examine the activation of mitogen activated protein kinase (MAPK) signaling pathways. Activator protein (AP) 1/DNA interactions were evaluated by electrophoretic mobility shift assay (EMSA). RESULTS: Northern blot showed that TNF-alpha stimulated CCL2 gene expression in NPFs. Significant increase of B-Raf, phosphorated MAPK including mitogen activated ERK-activate kinase (MEK)1/2, extracellular signal-related kinase 1/2, and p38 were detected by Western blot. c-Fos and c-Jun mRNAs were induced by TNF alpha, and PD98059 (MEK inhibitor) and SB203580 (p38 inhibitor) abolished the up regulation of c-Fos. EMSA revealed that TNF-a increased AP-1/DNA binding, and PD98059 and SB203580 attenuated this reaction, possibly via reducing c-Fos synthesis. PD98059 and curcunmin (AP-1 inhibitor) markedly suppressed the TNF alpha-induced CCL2 expression, whereas the effect of SB203580 was less noted. CONCLUSION: TNF-alpha induces CCL2 transcription in NPFs. B-Raf/MEK/ERK signaling cascade and to a less extent the p38 pathway are responsible for c-Fos activation and the subsequent AP-1/DNA interaction leading to CCL2 expression. PMID- 17424891 TI - Embracing change. Flexibility is key to design of suburban hospital. PMID- 17424892 TI - Incredible 'vistas'. ASHE and AIA/AAH salute best in teamwork for new construction, renovation and infrastructure. PMID- 17424893 TI - In sync. Basics of health facility commissioning. PMID- 17424894 TI - Traffic report. Advice on improving a hospital wayfinding system. PMID- 17424895 TI - Real numbers: understanding square footages and building multipliers that work. PMID- 17424896 TI - Fabulous fabrics: cleaning and spotting upholstered furnishings in the health care setting. PMID- 17424897 TI - Poor data threatens future of PbR system, warns minister. PMID- 17424898 TI - Call for joint posts to ease tension. PMID- 17424899 TI - Out-of-hours-care. The urgent care muddle may mean more nights to forget. PMID- 17424900 TI - On thought at last orders. PMID- 17424901 TI - Accountability. Listen and learn: keys to good commissioning. PMID- 17424902 TI - Data briefing. Patients opting for care close to home. PMID- 17424903 TI - Legislation. Power of two. AB - New legislation will place a duty on councils and PCTs to work together to assess health needs. It is hoped a new era of joint working can encourage councils and PCTs find overlaps and gaps in service. The government also hopes more joined-up working can help it shift care out of hospitals into the community. PMID- 17424904 TI - Payment by results. Major malfunction. PMID- 17424905 TI - Equality. Flexible friends. PMID- 17424906 TI - L1 elements, processed pseudogenes and retrogenes in mammalian genomes. AB - Long interspersed nuclear elements 1 (L1 elements or LINE1) are the most active autonomous retrotransposons in mammalian genomes. In addition to L1 elements themselves, other protein-coding mRNAs can also be reverse transcribed and integrated into the genome through the L1-mediated retrotransposition, leading to the formation of processed pseudogenes (PPs) and retrogenes, both of which are characterized by the lack of introns and the presence of a 3' polyA tract and flanking direct repeats. PPs are unable to encode a functional protein and have accumulated frameshift mutations and premature stop codons during evolution. A few of PPs are transcriptionally active. Retrogenes preserve undisrupted coding frames and are capable of encoding a functional protein that is identical or nearly identical to that of the progenitor gene. There is a significant excess of retrogenes that originate from the X chromosome and are retrotransposed into autosomes, and most of these retrogenes are specially expressed in male germ cells, suggesting the inactivation of X-linked genes during male meiosis provides a strong selection pressure on retrogenes originating from the X chromosome. PMID- 17424907 TI - Mitochondrial Abeta: a potential cause of metabolic dysfunction in Alzheimer's disease. AB - Deficits in mitochondrial function are a characteristic finding in Alzheimer's disease (AD), though the mechanism remains to be clarified. Recent studies revealed that amyloid beta peptide (Abeta) gains access into mitochondrial matrix, which was much more pronounced in both AD brain and transgenic mutant APP mice than in normal controls. Abeta progressively accumulates in mitochondria and mediates mitochondrial toxicity. Interaction of mitochondrial Abeta with mitochondrial enzymes such as amyloid beta binding alcohol dehydrogenase (ABAD) exaggerates mitochondrial stress by inhibiting the enzyme activity, releasing reactive oxygen species (ROS), and affecting glycolytic, Krebs cycle and/or the respiratory chain pathways through the accumulation of deleterious intermediate metabolites. The pathways proposed may play a key role in the pathogenesis of this devastating neurodegenerative disorder, Alzheimer's disease. PMID- 17424908 TI - Molecular mechanisms of calorie restriction's protection against age-related sclerosis. AB - The current knowledge on the molecular mechanisms of the protective effect of calorie restriction (CR) against age-related fibrosclerosis is tentatively reviewed with specific reference to the role of oxidative stress in aging. The effects of oxidative stress are often mediated by its own final products. Of these, 4-hydroxy-2,3-nonenal (HNE) induces the expression and synthesis of transforming growth factor beta1 (TGFbeta1) and activates nuclear binding of transcription factor activator protein 1 (AP-1) thus stimulating fibrogenesis. Several studies have shown that, as well as extending mean and maximum life span in a variety of species, CR delays the onset and slows the progression of a variety of age-associated diseases, including diabetes, cardiovascular diseases and neoplasia. However, the anti-aging mechanisms of CR are still not clearly understood. Of the numerous hypotheses put forward, one that still remains popular is protection against the age-associated increase of oxidative stress and consequent cell damage. CR protects the rat aorta from the age-related increase of both oxidative damage and fibrosis; as regards the possible mechanism/s of CR's protection against fibrosclerosis, it is conceivable that, by decreasing oxidative stress, CR reduces HNE levels and consequently TGFbeta1 expression and collagen deposition, likely by down-regulating the activation of Jun-N terminal kinase and of AP-1. Through the modulation of reactive oxygen species and oxidative stress CR may also attenuate the age-associated increase in the inflammatory milieu, thus preserving vascular functional integrity by suppressing the age-associated increase in inflammatory enzyme activities and prostanoids. PMID- 17424909 TI - Triosephosphate isomerase deficiency: facts and doubts. AB - Many glycolytic enzymopathies have been described that manifest clinically as chronic hemolytic anemia. One of these, triosephosphate isomerase (TPI) deficiency, is unique among the glycolytic enzyme defects since it is associated with progressive neurological dysfunction and frequently with childhood death. The physiological function of TPI is to adjust the rapid equilibrium between dihydroxyacetone phosphate and glyceraldehyde-3-phosphate produced by aldolase in glycolysis, which is interconnected to the pentose phosphate pathway and to lipid metabolism via triosephosphates. The TPI gene is well characterized; structure and function studies suggest that instability of the isomerase due to different mutations of the enzyme may underlie the observed reduced catalytic activity. Patients with various inherited mutations have been identified. The most abundant mutation is a Glu104Asp missense mutation that is found in homozygotes and compound heterozygotes. Two germ-line identical Hungarian compound heterozygote brothers with distinct phenotypes question the exclusive role of the inherited mutations in the etiology of neurodegeneration. This paper: (i) reviews our present understanding of TPI mutation-induced structural alterations and their pathological consequences, (ii) summarizes the consequences of TPI impairment in the Hungarian case at local and system levels, and (iii) raises critical questions regarding the exclusive role of TPI mutations in the development of this human disease. PMID- 17424910 TI - Steroid hormone rapid signaling: the pivotal role of S-palmitoylation. AB - Nuclear receptors (NRs) comprise a large family of proteins essential to manipulate various aspects of human biology. The class III of the NR family includes steroid hormone receptors (SHRs) involved in embryonic development, maintenance of differentiated cellular phenotypes, metabolism, and cell death. Dysfunction of SHR signaling leads to proliferative, reproductive, and metabolic diseases. SHRs are ligand-activated transcription factors which regulate cellular processes by inducing genomic events. In addition, membrane-initiating non genomic signals have been described for several SHRs. These non-genomic actions are independent of the SHR transcription activity and have been attributed to canonical SHRs and non-canonical hormone binding sites located at the plasma membrane. The molecular bases for SHR-plasma membrane association represent a debated issue. Here, we hypothesize that the SHR amino acid sequence homologous to that encompassing the S-palmitoylated Cys447 residue of human estrogen receptor alpha could represent a new consensus sequence for SHR S-palmitoylation. Reversible S-palmitoylation of SHRs could play a critical role in receptor localization at the plasma membrane and in rapid nongenomic signaling activation. PMID- 17424911 TI - Thermodynamic studies on the interaction of water-soluble porphyrins with the glucose/mannose-specific lectin from garden pea (Pisum sativum). AB - Due to the application of porphyrins as photosensitizers in photodynamic therapy to treat cancer, and the ability of some lectins to preferentially recognize tumor cells, studies on the interaction of porphyrins with lectins are of considerable interest. Here we report thermodynamic studies on the interaction of several free-base and metallo-porphyrins with pea (Pisum sativum) lectin (PSL). Association constants (Ka) were obtained by absorption titrations by monitoring changes in the Soret band of the porphyrins and the Ka values obtained for various porphyrins at different temperatures are in the range of 1.0 x 10(4) to 8.0 x 10(4) M(-1). Both cationic and anionic porphyrins were found to bind to PSL with comparable affinity. Presence of 0.1 M methyl-alpha-D-mannopyranoside--a carbohydrate ligand that is specifically recognised by PSL--did not affect the binding significantly, suggesting that porphyrin and sugar bind at different sites on the lectin. From the temperature dependence of the Ka values, the thermodynamic parameters, change in enthalpy and change in entropy associated with the binding process were estimated. These values were found to be in the range: delthaH degree = -95.4 to -33.9 kJ x mol(-1) and deltaS degree = -237.2 to -32.2 J x mol(-1) x K(-1), indicating that porphyrin binding to pea lectin is driven largely by enthalpic forces with the entropic contribution being negative. Enthalpy-entropy compensation was observed in the interaction of different porphyrins to PSL, with the exception of meso-tetra-(4 sulfonatophenyl)porphyrinato zinc(II), emphasizing the role of water structure in the overall binding process. Circular dichroism and differential scanning calorimetric studies indicate that while porphyrin binding does not induce significant changes in the lectin structure and thermal stability, carbohydrate binding induces moderate changes in the tertiary structure of the protein and also increases its thermal unfolding temperature and the enthalpy of the unfolding transition. PMID- 17424912 TI - Zinc fingers are known as domains for binding DNA and RNA. Do they also mediate protein-protein interactions? PMID- 17424913 TI - Revenue integrity departments support the revenue cycle. PMID- 17424914 TI - Scripts help collections staff. PMID- 17424915 TI - Analyze your costs in 3-D. PMID- 17424916 TI - [Pathophysiological and clinical significance of microalbuminuria]. PMID- 17424917 TI - [Nutritional status of peritoneal dialysis patients and parameters of dialysis adequacy]. AB - The aim of our study was to evaluate relations between peritoneal dialysis (PD) adequacy and nutritional parameters of PD patients. Patients (n = 124), who finished PD treatment, were separated on 2 groups according to the mean total Kt/V for the entire PD course being below 2.0 (group I) or over 2.0 (group II). Adequacy parameters, daily intake of food products and nutritional indices were evaluated in each patient every 3-6 months during the entire PD course. Mean values of examined parameters were used for comparison of differences observed between both groups. Group I included 63 men, 16 women, age 50.3 +/- 13.8 years, PD duration 13.2 +/- 10.1 months. Group II consisted of 12 men, 33 women, age 49.1 +/- 14.9 years, PD duration 8.8 +/- 6.0 months. Due to a significant difference in sex distribution, dialysis duration and ideal body mass (IBM) between groups, statistical analysis was performed with adjustment of results to these parameters. Absolute amounts of daily intake of food components were higher in group I for animal protein, sodium, retinol, niacin, saturated and polyunsaturated fatty acids and cholesterol. When daily food intake was normalized to IBM, group II showed higher both protein nitrogen appearance (I - 0.92 +/- 0.25, II - 1.12 +/- 0.45 g/kg IBM, p = 0.005) and intake of vegetable protein (I - 0.29 +/- 0.10, II - 0.34 +/- 0.09 g/kg IBM, p = 0.040), carbohydrates (I - 3.30 +/- 1.08, II - 3.80 +/- 1.34 g/kg IBM, p = 0.029), potassium (I - 33.2+/-10.6, II - 38.3 +/- 13.2 mg/kg IBM, p = 0.034), calcium (I 5.81 +/- 2.46, II - 7.20 +/- 3.54 mg/kg IBM, p = 0.028), magnesium (I - 2.86 +/- 0.86, II - 3.41 +/- 1.36 mg/kg IBM, p = 0.004), beta-carotene (I 22.4 +/- 15.8, II - 34.9 +/- 29.1 mg/kg IBM, p = 0.002) as well as calorie delivered from protein (I - 0.22 +/- 0.04, II - 0.26 +/- 0.07%, p = 0.001) and carbohydrates (I 0.79 +/- 0.15, II - 0.94 +/- 0.21, p = 0.000) in relation to total amount of ingested calorie. Group I showed significantly lower serum levels of albumin (I - 2.45, 1-3, II - 2.83, 1-3 scores, p = 0.023) and cholesterol (I - 5.54 +/- 1.06, II - 6.35 +/- 1.63 mmol/l, p = 0.009), but higher serum iron (I - 16.7 +/- 4.4, II - 15.8 +/- 5.2 micromol/l, p = 0.042) and ferritin (I - 615, 28-5113, II - 377, 24-3376 ng/ml, p = 0.021) concentrations as well as transferrin saturation (I - 31.1 +/- 9.2, II - 28.5 +/- 9.2%, p = 0.032). We conclude that PD patients with Kt/V over 2.0 as compared to those with Kt/V below 2.0 show tendency for better nutritional indices excluding serum iron parameters. PMID- 17424918 TI - [Interrelation between arterial hypertension and body mass index in patients with concomitant ischemic heart disease treated with cardiac rehabilitation]. AB - A goal of the study was to investigate the interrelations between body mass index (BMI) and arterial pressure level and its alterations induced by a long-term cardiac rehabilitation. In the patients with arterial hypertension with concomitant ischemic heart disease (IHD). The study group consisted of 73 patients (46 males and 27 females), with age range 54-64 years (the mean: 59.59 +/- 0.67 years), subjected to the 6-month ambulatory supervised cardiac rehabilitation, which comprised stamina training with cycloergometer combined with generally improving exercises. Arterial pressure level was measured in the resting state (just before the training onset) and at the interval training peak (peak pressure) during the consecutive three initial and three final training sessions. Upon completion of the rehabilitation cycle, statistically significant reduction of resting systolic pressure: 116.85 +/- 1.68 mmHg vs 105.12 +/- 1.43 mmHg (p<0.01) and insignificant reduction of resting diastolic pressure 79.85 +/- 1.24 mmHg vs 77.44 +/- 1.21 mmHg (NS) were observed. Additionally, insignificant lowering both systolic and diastolic peak pressure was noted, respectively: 119.45 +/- 1.92 vs 118.75 +/- 2.3 mmHg and 78.63 +/- 1.22 mmHg vs 76.96 +/- 1.30 mmHg. Correlation analysis concerning the values of resting systolic and diastolic pressure, likewise of peak systolic and diastolic pressure, on the one hand, and the values of body mass index, on the other hand, yielded in the all considered cases the statistically insignificant positive Pearson's coefficients. The further dependence analyzed, between the delta of resting and peak pressure, systolic and diastolic, and the body mass index values was featured by the insignificant statistically negative correlation coefficients. In result of the 6 month rehabilitation, 36 out of 73 patients demonstrated BMI value reduction by 0.65 +/- 0.33, which did not affect significantly an extent of the arterial pressure lowering. Therefore, in the examined patient group with hypertension and IHD, body mass index turned out to determine neither the initial nor the after rehabilitation pressure levels, both systolic and diastolic. PMID- 17424919 TI - [The plasma levels of granulocyte-colony stimulating factor (G-CSF) and macrophage-colony stimulating factor (M-CSF) in breast cancer patients]. AB - We have investigated the plasma levels of G-CSF and M-CSF and commonly accepted tumor marker CA 15-3 in 54 breast cancer patients before and after surgery and chemotherapy. The patients were divided into two groups: A (stage I) and B (stage II). G-CSF i M-CSF were determined using enzyme-linked immunosorbent assay (ELISA), CA 15-3 was measured by microparticle enzyme immunoassay (MEIA). G-CSF and M-CSF plasma levels (similarly as CA 15-3) were significantly higher in breast cancer patients before surgery comparing to the control group. After surgery plasma level of G-CSF (as CA 15-3) was decreased, but M-CSF increased. The plasma levels of tested cytokines and CA 15-3 increased after chemotherapy. This study suggests that tested cytokines (especially G-CSF) can be clinically useful in diagnostics of breast cancer, yet further investigation and confirmation by a prospective study are necessary. PMID- 17424920 TI - [Distribution of extrema on body surface isointegral maps in RBBB]. AB - The investigations were undertaken in order to assess the characteristic features of the isointegral maps obtained from the patients demonstrating on the standard ECG complete and incomplete right bundle branch block (RBBB). The body surface potential mapping (BSPM) procedure was performed using Fukuda Denshi system enabling to record electrocardiographic signals simultaneously from the 87 leads arranged over anterior and posterior thoracic surfaces. The isointegral maps of QRST, QRS, early portion of QRS complex (from QRS onset to R wave peak), and of late portion of QRS (from R wave peak to QRS offset) were created. The distribution of extrema (maxima and minima) was found to be specific for RBBB cases and the characteristic differences between RBBB group and the controls were observed. PMID- 17424921 TI - [Diabetes control in type 2 diabetic subjects treated in primary health care]. AB - Aim of the study was to assess control of type 2 diabetes in subjects treated by general practitioners. Study was conducted in one of the primary health care centers in a big city, in which health care was provided for 27 900 inhabitants. Control of diabetes was assessed in 355 of all 936 type 2 diabetic subjects registered in the center. None of them was seen by diabetologist in the preceeding year. Mean age was 65,7 +/- 10,3 lat, diabetes duration 9,2 +/- 7,3 lat, BMI 29,9 +/- 4,9 kg/m2. Normal body weight was found in 15%, overweight in 39%, obesity in 46% of subjects. Hypertension was present in 81%, dyslipidaemia in 62% of patients. Mean HbAlc was 7,2 +/- 1,3%, fasting serum glycaemia 144 +/- 48 mg/dl, total cholesterol 204 +/- 45 mg/dL, LDL - 119 +/- 33 mg/dL, HDL - 51 +/ 13 mg/dl, triglicerides 182 +/- 108 mg/dL. Mean systolic blood pressure was 146 +/- 20 mmHg, diastolic 83 +/- 11 mmHg. Treatment goals recommended by Polish Diabetological Association in 2005 were attained as follows: HbAlc < or = 6,1% - 19,7% of subjects, fasting glycaemia < or =110 mg/dl - 24%, total cholesterol < 175 mg/dl - 26%, LDL < 100 mg/dl - 29%, triglicerides < 150 mg/dl - 46%, cholesterol HDL > 40 mg/dl in men and > 50 mg/dl in women - 65% of subjects. Recommended systolic blood pressure < 130 mmHg was found in 16 %, diastolic blood pressure < 80 mmHg - in 24%, and both values - in 8% of diabetics. In no one subject all recommended treatment goals were met. Conclusions 1. Recommended treatment goals are perceived in unacceptably low number of type 2 diabetic subjects treated by general practitioners. 2. Medical care of type 2 diabetic subjects performed in primary health care is unsatisfactory and should be essentially improved or changed. PMID- 17424922 TI - [Omalizumab in severe asthma--case report]. AB - We describe the case of 31-year-old woman whose severe asthma was not properly controlled with best available therapy according to GINA (2002). Following administration of anti-IgE monoclonal antibody she experienced significant improvement in her asthma control PMID- 17424923 TI - [Case report of patient with systemic lupus erythematosus proceed with lymphadenopathy]. AB - On the basis of observed case we deciding describe the diagnostic problems connected with atypical course of systemic lupus erythematosus (SLE) with accompanying of lymphadenopathy and viral infection. In the differentiation diagnostic beside hematologic malignancies and SLE we take into consideration also other diseases, which may be manifested by lymphadenitis, such as tuberculosis, sarcoidosis, metastasis, Kikuchi-Fujimoto disease and Kawasaki disease. Corticosteroid treatment cause improvement of clinic condition of subject and standardization of many laboratory results. PMID- 17424924 TI - [Diagnostic problems in eosinophilic fasciitis]. AB - We presented two cases with symptoms of diffuse swelling of subcutaneous tissue, stiffness and tenderness of involved areas, fever, eosinophilia and hypergammaglobulinemia. The inflammatory infiltrates consisting of lymphocytes, plasma cells and eosinophils were yielded in fascia. The difficulties in differentition of the symptoms between eosinophilic fasciitis and "eosinophilia myalgia syndrome" are discussed. PMID- 17424925 TI - [Contemporary view on subclinical hyperthyroidism]. PMID- 17424926 TI - [Rimonabant--a panacea for patients with metabolic syndrome?]. PMID- 17424927 TI - [The effect of ethanol on the protein glycosylation. Part 1. Microheterogeneity of serum glycoproteins]. PMID- 17424928 TI - [Treatment of obstructive sleep apnea]. PMID- 17424929 TI - Metabolites from the fungus Cephalosporium sp. AL031. AB - A new pyrone derivative, 7, 9-dihydroxy-10-methyl-2H, 4aH, 6H, 10bH-pyrano[5,6 c][2]ben-zopyran-2,6-dione (1), was isolated from a culture broth of a strain of the fungus Cephalosporium sp. AL031, together with three known compounds, 3 acetyl-7-hydroxy-5-methoxyl-3H-isobenzofuran-1-one (2), vermopyrone (3), and 5 methylmellein (4). Their structures were elucidated by spectroscopic analysis including MS and 2D-NMR. Compounds 2, 3, and 4 are reported for the first time from fermentation broth of this fungus through the present study. PMID- 17424930 TI - Two indolocarbazole alkaloids with apoptosis activity from a marine-derived actinomycete Z(2)039-2. AB - Bioassay-guided fractionation of the EtOAc extract from the fermentation broth of a marine-derived actinomycete Z(2)039-2 led to the isolation of two known indolocarbazole alkaloids, K252c (1) and arcyriaflavin A (2). 1 and 2 exhibited moderate cytotoxic activities against the K562 cell line, and induced apoptotic activities at 10 and 100 microM, respectively. This is the first report on the significant apoptosis inducing effect of indolocarbazole alkaloids against K562 cancer cells. PMID- 17424931 TI - Antioxidant activity of cleomiscosins A and C isolated from Acer okamotoanum. AB - Phytochemical investigation of Acer okamotoanum leaf and twig led to the isolation of two coumarinolignans, cleomiscosin A (1) and cleomiscosin C (2). Here, we found that 2 dose-dependently inhibits LDL oxidation mediated by either catalytic copper ions (Cu2+) or free radicals generated with the azo compound 2,2'-azobis-(2-amidinopropane)dihydro-chloride (AAPH) with IC50s of 29.5 and 11.9 microM, respectively. By electrophoretic analysis, we also observed that 2 protects apolipoprotein B-100 (apoB-100) against Cu2+-induced fragmentation (65.3% inhibition at 5 microM). Furthermore, fluorescence analyses clearly indicated that both 1 and 2 protect against the oxidative modification of apoB 100 induced by either Cu2+ or HOCl (1, IC50s of 13.4 and 8.1 microM, respectively; 2, IC50s of 23.6 and 3.9 microM, respectively). These findings suggest that 1 and 2 could be beneficial in preventing LDL oxidation in atherosclerotic lesions. PMID- 17424932 TI - Modification of C2 functional group on asiatic acid and the evaluation of hepatoprotective effects. AB - For the development of novel hepatoprotective agents, C2 functional group on asiatic acid was modified, and evaluated for their hepatoprotective effects. Among prepared compounds, compounds 10 and 14 showed better hepatoprotective effects compared to silymarin. PMID- 17424933 TI - Structure activity relationship of antioxidative property of flavonoids and inhibitory effect on matrix metalloproteinase activity in UVA-irradiated human dermal fibroblast. AB - Collagenase, a matrix metalloproteinases (MMPs), is a key regulator in the photoaging process of skin due to the reactive oxygen species generated after exposure to ultraviolet A (UVA). Flavonoid compounds have been demonstrated to possess antioxidant properties, and could be useful in the prevention of photoaging. In this study, to investigate the structure-activity relationship of flavonoid compounds on their antioxidant property and inhibitory effects against the MMP activity, the effects of several flavonoids; myricetin, quercetin, kaempferol, luteolin, apigenin and chrysin, on the reactive oxygen species scavengering activity and inhibitory effect against the MMP activity were examined in vitro and in human dermal fibroblasts induced by UVA. The relative order of antioxidative efficacy, as determined using the 1, 1-diphenyl-2 picrylhydrazyl (DPPH) method and the xanthine/xanthine oxidase system, was as follows; flavones: luteolin > apigenin > chrysin, flavonols: myricetin > quercetin > kaempferol, and correlated with the respective number of OH group on their B-ring. In good correlation with the antioxidant properties, the flavonoids inhibited the collagenase activities, in a dose-dependent manner, and the MMP expression. These results suggested the UVA induced antioxidative activity and inhibitory effects of flavonoids on the collagenase in human dermal fibroblasts depends on the number of OH group in the flavonoid structure, and those with a higher number of OH group may be more useful in the prevention of UV stressed skin aging. PMID- 17424934 TI - Farnesyl protein transferase and tumor cell growth inhibitory activities of lipiferolide isolated from Liriodendron tulipifera. AB - The methanolic extract of the leaves of Liriodendron tulipifera was found to show inhibitory activity towards farnesyl protein transferase (FPTase). Bioassay guided fractionation of the methanolic extract resulted in the isolation of lipiferolide, an inhibitor of FPTase. This compound inhibited the FPTase activity in a dose-dependent manner, and showed cell growth inhibitory activity against several tumor cells. PMID- 17424935 TI - The analgesic, anti-inflammatory and calcium antagonist potential of Tanacetum artemisioides. AB - Several species of the genus Tanacetum are traditionally used in a variety of health conditions including pain, inflammation, respiratory and gastrointestinal disorders. In the current investigation, we evaluated the plant extract of T. artemisioides and some of its pure compounds (flavonoids) for analgesic, anti inflammatory and calcium antagonist effects in various in-vivo and in vitro studies. Using the actetic acid induced writhing test, intraperitoneal (i.p) administration of the plant extract (25-50 mg/kg) and its flavonoid compounds TA 1 and TA-2 (1-5 mg/kg ) exhibited significant analgesic actvity. The maximum analgesic effect observed with the crude extract of the plant was 71% at 50 mg/kg, while that of compounds TA-1 and TA-2 (5 mg/kg i.p) was 75 and 47%, respectively. The plant extract and its pure compounds caused inhbition of formalin induced paw licking in mice predominatly in the second phase of the test. Diclofenac sodium, a standard reference compound, showed a simlar effect in these chemical induced pain models. In the carrgeenan induced rat paw edema assay, the plant extract (50-200 mg/kg i.p) demonstrated significant (P< 0.01) anti-inflammatory activity which was comparable to that obtained with diclofenac sodium and indomethacin. In isolated rabbit jejunum preprations the plant extract showed an atropine sensitive dose-dependent (0.10-1.0 mg/mL) spasmogenic activity followed by a spasmolytic effect at the next higher doses (3-5 mg/mL). The crude extract of the plant also inhibited the high K+-induced contractions, indicating a calcium channel blocking (CCB) activity, which was further confirmed when the plant extract caused a rightward shift in the Ca++ concentration response curves in the isolated rabbit jejunum preparations, similar to that seen with verapamil. The flavonoid compounds isolated from the plant were devoid of any activity in the isolated tissue preparations. These results indicate that the plant extract of T. artemisioides possesses analgesic, anti-inflammatory and CCB activities. The flavonoid compounds of the plant may have a role in its observed analgesic and antiinflammatory activities, while the CCB activity of the plant may be attributed to some other chemical constituents present. Moreover the findings support the traditional reputation of the genus Tanacetum for its therapeutic benefits in pain and inflammatory conditions. PMID- 17424936 TI - Topoisomerase I inhibition and cytotoxicity of licochalcones A and E from Glycyrrhiza inflata. AB - Licochalcones A (1) and E (2), retrochalcones or reversely constructed chalcones, isolated from the roots of Glycyrrhiza inflata, were evaluated for their cytotoxicities against four different human tumor cell lines; A549 (lung), SK-OV 3 (ovarian), SK-MEL-2 (melanoma) and HCT-15 (colon), using the sulforhodamine B (SRB) assay. The effects of these compounds toward the DNA topoisomerase I (topo I) inhibitory activity were also measured using the supercoiled DNA unwinding assay. All compounds showed moderate cytotoxicities against the four different human tumor cell lines and inhibited the topo I activity in dose-dependent manners. The inhibition of topo I by licochalcones A (1) and E (2) may explain the cytotoxicities of these compounds against the human tumor cell lines. PMID- 17424937 TI - Endogenous sphingolipid metabolites related to the growth in Sphingomonas chungbukensis. AB - Sphingolipids are present in animals, plants, fungi, yeasts and some bacteria. In mammalian cells sphingolipids act as lipid mediators for cell growth, differentiation, apoptosis and angiogenesis. In contrast, in bacteria the biological significance of sphingolipids has not been fully elucidated and sphingolipid metabolism has not been investigated. The aim of this study was to compare the pattern of sphingolipid metabolites in HIT-T15 beta cells originating from hamster pancreas to that in the bacterial strain Sphingomonas chungbukensis DJ77, under various culture conditions. It was found that the concentration of cellular sphinganine (Sa) in S. chungbukensis was higher than that of sphingosine (So), while the level of cellular So in HIT-T15 cells was higher than that of Sa. Aeration and shaking during culture increased bacterial growth in S. chungbukensis, and the contents of So and Sa were also elevated. These results indicate that a de novo sphingolipid pathway appeared to be active in bacteria and that bacterial growth may be closely related to Sa levels. PMID- 17424938 TI - Effects of beta-glucan from Aureobasidium pullulans on acute inflammation in mice. AB - The effects of beta-glucan isolated from Aureobasidium pullulans were observed on acute xylene-induced inflammation. beta-glucan at a dose of 62.5, 125 or 250 mg/kg were administered once orally to xylene-treated mice (0.03 mL of xylene was applied on the anterior surface of the right ear to induce inflammation), and the body weight change, ear weight, histological profiles and histomorphometrical analyses of ear were conducted upon sacrifice. The xylene was topically applied 30 min after dosing with beta-glucan. The results were compared to those of diclofenac, indomethacin and dexamethasone (15 mg/kg injected once intraperitoneally). All animals were sacrificed 2 h after xylene application. Xylene application resulted in marked increases in induced ear weights compared to that of intact control ear; hence, the differences between intact and induced ear were also significantly increased. The histological characteristics of acute inflammation, such as severe vasodilation, edematous changes of skin and infiltration of inflammatory cells, were detected in xylene-treated control ears with marked increase in the thickness of the ear tissues. However, these xylene induced acute inflammatory changes were significantly and dose-dependently decreased by beta-glucan treatment. We conclude that beta-glucan from A. pullulans has a somewhat favorable effect in the reduction of the acute inflammatory responses induced by xylene application in mice. PMID- 17424939 TI - Occurrence of the van genes in Enterococcus faecalis and Enterococcus faecium from clinical isolates in Korea. AB - Enterococcus faecalis and Enterococcus faecium isolated from Korea patients in 1998 and 2005 were tested for susceptibility to nine different antimicrobial agents, including vancomycin and teicoplanin. Polymerase chain reaction (PCR) was used for the detection of several vancomycin resistance genes such as vanA ('high level'), vanB ('moderate high level'), vanC1 and vanC2 ('low level'). Both E. faecalis and E. faecium exhibited a resistance of 80% and more than 60% to synercid and mupirocin, respectively. Moreover, an average of 76% of all isolates was resistant to ciprofloxacin, gentamicin, lincomycin, cefotaxime, and meropenem, confirming the multiple drug resistance of most of the isolates. No resistance to vancomycin or teicoplanin was observed in the 1998 E. faecalis and E. faecium isolates. However, the 2005 E. faecalis and E. faecium isolates exhibited resistance of 16% and 12% to vancomycin and teicoplanin, respectively. In addition, vanA gene was detected in 4 strains of 2005 E. faecium isolates, thus showing a high resistance to vancomycin. No other vancomycin resistance genes, including vanB, vanC1, and vanC2, were found in our isolates. In this study, we compared the occurrence of antimicrobial resistance and the presence of specific resistance genes in E. faecalis and E. faecium. The results showed that the 2005 isolates were usually more resistant than the 1998 isolates. PMID- 17424940 TI - Inhibitory effect of bisphenol A on gap junctional intercellular communication in an epithelial cell line of rat mammary tissue. AB - An endocrine disruptor, bisphenol-A (BPA), has been reported to have several short-term actions in various cells and tissues. However, the mechanisms of these actions have not been fully elucidated. In order to assess the effect of BPA on the intercellular communication mediated by gap junctions, we conducted the present study in the rat epithelium-derived BICR-M1Rk cell-line, in which connexin 43 (Cx43) is a major gap junction channel-forming protein. The cytotoxicity of BPA toward the cultured cells was evaluated by using both MTT reduction and LDH leakage assay systems. The results showed no appreciable loss in cell viability in the presence of increasing concentrations of BPA (from 0.1 to 3.2 microM) for 1 h incubation. However, most of cell viability was lost when cells were incubated for 24 hr with the same concentrations of BPA. The BPA acted as an antagonist on gap junction-mediated intercellular communication (GJIC), and the phenomenon was dose-dependent and irreversible. According to the data obtained from scrape-loading dye-transfer experiments, three quarters of normal GJIC was reduced by concentration of 0.4 microM BPA for 1 h incubation. To identify the relevance of this retardation upon BPA treatment, the GJIC to Cx43 synthesis, the mRNA and protein levels of Cx43 were assessed with RT-PCR and Western-blotting, respectively. The total protein level of Cx43 was almost constant in a wide range of BPA concentrations, as well as in Cx43 mRNA level. These results suggest that BPA inhibits GJIC through a modulation of the gating of gap junction channels, not through a genomic modulation of Cx43. PMID- 17424941 TI - CYP2C19 polymorphism in Korean patients on warfarin therapy. AB - This study was designed to assess the effect of CYP2C19 polymorphism on warfarin dosage requirements and bleeding complications in the Korean population. Patients were placed into one of four groups according to the dose of warfarin they received and the presence of bleeding complications: regular dose control, regular dose bleeding, low dose control, and low dose bleeding. Genotyping for CYP2C19*2 and CYP2C19*3 was performed by the restriction fragment length polymorphism method for each patient and each study group. The measured internal normalized ratio (INR) in each dose group was similar even though the administered dosage was significantly different. A total of 66 patients were evaluated for CYP2C19 polymorphism. Among them 25 patients (37.9%) were homozygous wild type. Four patients (6.1%) had heterozygous mutations at both loci. Others had mutations on either the CYP2C19*2 or *3 locus. Higher genetic variation was observed in CYP2C19*2 than in CYP2C19*3 among Korean patients on warfarin therapy. Our data suggested that there is a higher incidence of bleeding complications in patients who have a higher allele frequency of CYP2C19. It was also revealed that the distribution of CYP2C19 polymorphism among Asian populations is more similar than of the distribution among Caucasian populations. PMID- 17424942 TI - Inhibition of mouse macrophages interleukin-12 production: suppression of nuclear factor-kappaB binding activity by a specific factor isolated from Scapharca broughtonii. AB - Pharmacological inhibition of interleukin-12 (IL-12) production may allow a therapeutic strategy for preventing development and progression of disease in experimental models of autoimmunity. In this study we investigated the effects of an ethanol fraction of the Scapharca broughtonii, on the production of IL-12 by mouse macrophages stimulated with lipopolysaccharides (LPS). The ethanol fraction (S3) prepared from Scapharca broughtonii potently inhibited LPS-induced IL-12 production in the RAW264.7 monocyte cell-line in a dose-dependent manner. The activation effect of the ethanol fraction (S3) on the IL-12 gene promoter was analyzed by transfecting RAW264.7 cells with IL-12 gene promoter/luciferase constructs. The repressive effect mapped to a region in the IL-12 gene promoter that contained a binding site for NF-kappaB. Furthermore, activation of macrophages by LPS resulted in markedly enhanced binding activity to the NF kappaB site, which significantly decreased upon addition of the ethanol fraction, indicating that the ethanol fraction of the blood shell inhibited IL-12 production in LPS-activated macrophages via inhibition of NF-kappaB binding activity. PMID- 17424943 TI - In vivo kinetics and biodistribution of HB-110, a novel HBV DNA vaccine, after administration in mice. AB - This study investigated the pharmacokinetic profile and biodistribution of HB 110, a novel HBV therapeutic vaccine candidate, in mice. HB-110 was rapidly degraded in the blood after i.v. injection with a half-life of 1.9+/-0.083 min, and was no longer detected at 60 min except in one individual near the detection limit. In the i.m. injection, plasmid DNA was detectable at the injection site until 11 days after administration, but the amounts were just above the detection limit. The blood concentration of HB-110 showed a maximum of 604 pg/mL at 15 min after i.m. injection, which was followed by degradation to undetectable levels at 90 min. The plasmid DNA in tissues peaked at 90 min after administration. The highest concentration of plasmid DNA was detected in the liver (24.172 pg/mg tissue), and considerable amounts were also observed in the lung (9.467 pg/mg tissue) and spleen (7.688 pg/mg tissue). The amount of plasmid DNA in tissues was 2 to 3 orders of magnitude lower than in the injection site at the same time points. The HB-110 concentration in tissues, including gonads, decreased rapidly and was undetectable 24 h after administration. PMID- 17424944 TI - Isolation of fatty acids with anticancer activity from Protaetia brevitarsis larva. AB - In this study, biologically active compounds were isolated from Protaetia brevitarsis larva (PBL) by dichloromethane extraction. The dichloromethane extract from PBL was highly cytotoxic to various cancer cells. From a silica gel column chromatograpy of this extract, we obtained four fractions (F-2, F-4, F-5 and F-7) having apoptosis-inducing activity. These fractions induced DNA ladder and caspase-3 activation during apoptosis in colon 26 tumor cells. In 1H and 13C NMR and mass spectral analysis of the fraction F-2 showing the highest apoptosis inducing activity, we found that the fraction was composed of three free fatty acids such as palmitic acid, (Z)-9-octadecenoic acid and octadecenoic acid. These results indicate that the dichloromethane extract of PBL includes anticancer components composed of at least three fatty acids, and apoptosis-inducing activity of the extract was mediated by caspase-3 activation in tumor cells. PMID- 17424945 TI - Intranasal vaccination with peptides and cholera toxin subunit B as adjuvant to enhance mucosal and systemic immunity to respiratory syncytial virus. AB - Amino acid sequences 200-225 and 255-278 of the F protein of human respiratory syncytial virus (HRSV) are T cell epitopes (Bourgeois et al., 1991; Corvaisier et al, 1993). Peptides corresponding to these two regions were synthesized and coupled with keyhole limpet haemocyanin (KLH). The two conjugated proteins were administered intranasally to BALB/c mice alone or together with cholera toxin B (CTB). ELISAs revealed that the mixture of the conjugates with CTB increased not only the systemic response but also the mucosal immune response of the saliva. The systemic response was lower and the mucosal immune response was undetectable in mice immunized with the conjugates on their own. These results suggest that these two peptide sequences are effective epitopes for inducing systemic and mucosal immune responses in conjunction with CTB, and may provide the basis for a nasal peptide vaccine against RSV for human use. PMID- 17424946 TI - Effects of guanosine on the pharmacokinetics of acriflavine in rats following the administration of a 1:1 mixture of acriflavine and guanosine, a potential antitumor agent. AB - Preclinical studies are currently underway to examine the potential antitumor effects of a 1:1 mixture of acriflavine (ACF; CAS 8063-24-9) and guanosine. Guanosine potentiates the anticancer activity of some compounds. However, the effects of guanosine on the pharmacokinetics of ACF in mammals are unknown. Therefore, this study investigated the effects of guanosine on the pharmacokinetics of ACF after administering a 1:1 mixture of ACF and guanosine in rats. The rats were given either 10 mg/kg of the mixture or 5 mg/kg ACF via an intravenous bolus injection; or 30 mg/kg of the mixture or 15 mg/kg ACF intramuscularly. An HPLC-based method, which was validated in this laboratory, was used to analyze the levels of trypaflavine (TRF) and proflavine (PRF) in the plasma, bile, urine, and tissue homogenates. It was found that TRF and PRF were rapidly cleared from the blood and transferred to the tissues after the i.v. bolus or i.m. injection of the combination mixture. Both TRF and PRF were found to be most highly concentrated in the kidneys after the i.v. bolus or i.m. injection, followed by slow excretion to the bile or urine. Guanosine had no effect on the plasma disappearance of TRF or PRF after the i.v. bolus injection. However, guanosine led to a prolongation of the plasma levels of PRF after the i.m. administration of the combination mixture, resulting in a 2 fold increase in the bioavailability (BA) of PRF The concentrations of TRF and PRF in all the tissues examined were similar in the groups given the mixture and ACF. However, guanosine led to a prolongation of the biliary and urinary excretions of both TRF and PRF after the i.v. bolus (1.25 fold) or i.m. (1.5-2.4 folds) injection. These prolonged effects of guanosine on the plasma disappearance or urinary excretion of TRF and PRF might be one reason for the enhanced antitumor effects of ACF. However, more study will be needed to further examine this potential mechanism. PMID- 17424947 TI - A novel mucoadhesive polymer film composed of carbopol, poloxamer and hydroxypropylmethylcellulose. AB - Using the casting method novel mucoadhesive polymer blend film consisting of Carbopol, poloxamer, and hydroxypropylmethylcellulose (HPMC) was prepared and characterized. Triamcinolone acetonide (TAA) was loaded into Carbopol/poloxamer/HPMC polymer blend film. Carbonyl band of Carbopol in Carbopol/poloxamer/HPMC shifted to longer wavenumber than that of Carbopol in Carbopol/poloxamer due to the hydrogen bonding among Carbopol, poloxamer, and HPMC. Tan delta peak assigned to glass transition temperature (Tg) of HPMC shifted to low temperature due to increased flexibility caused by increased poloxamer content in polymer blend films. Swelling ratio of Carbopol/poloxamer/HPMC films was lowest in Carbopoll poloxamer/HPMC at mixing ratio of 35/30/35 (wt/wt/wt). Adhesive force of Carbopol/poloxamer/HPMC films increased with increasing HPMC content in Carbopol/poloxamer/HPMC polymer blend film and increasing hydroxypropyl group content in HPMC due to hydrophobic property of HPMC although bioadhesive force was highest at mixing ratio of 35/30/35 (wt/wt/ wt). Release of TAA from TAA-loaded Carbopol/poloxamer/HPMC polymer blend film in vitro increased with increasing loading content of drug. PMID- 17424948 TI - Two step derivatization for the analyses of organic, amino acids and glycines on filter paper plasma by GC-MS/SIM. AB - A rapid dried-filter paper plasma-spot analytical method was developed to quantify organic acids, amino acids, and glycines simultaneously in a two-step derivatization procedure with good sensitivity and specificity. The new method involves a two-step trimethylsilyl (TMS) - trifluoroacyl (TFA) derivatization procedure using GC-MS/ selective ion monitoring (GC-MS/SIM). The dried-filter paper plasma was fortified with an internal standard (tropate) as well as a standard mixture of distilled water and methanol. Methyl orange was added to the residue as an indicator. N-methyl-N-(trimethylsilyl-trifluoroacetamide) and N methyl-bis-trifluoroacetamide were then added and heated to 60 degrees C for 10 and 15 min to produce the TMS and TFA derivatives, respectively. Using this method, the silylation of carboxylic functional groups was carried out, which was followed by the trifluoroacyl derivatization of the amino functional group. The derivatives were analyzed by GC-MS/SIM. A calibration cure showed a linear relationship for the target compounds between concentrations of 10-500 ng/mL. The limit of detection and quantification on a plasma spot were 10-90 ng/mL (S/N=9) and 80-500 ng/ mL, respectively. The correlation coefficient ranged from 0.938 and 0.999. When applied to the samples from positive patients, the method clearly differentiated normal subjects from the patients with various metabolic disorders such as PKU, MSUD, OTC and a Propionic Aciduria. The new developed method might be useful for making a rapid, sensitive and simultaneous diagnosis of inherited organic and amino acid disorders. In addition, this method is expected to be an alternative method for screening newborns for metabolic disorders in laboratories where expensive MS/MS is unavailable. PMID- 17424949 TI - Integrating safety into Kaizen events. PMID- 17424951 TI - Extending your leadership reach: making central and local connections. PMID- 17424950 TI - Silent but deadly. PMID- 17424952 TI - Break right!! How to survive the missiles of life. PMID- 17424953 TI - The long debate over belt air. PMID- 17424954 TI - Performing the lockout/tagout risk assessment. AB - Lockout/tagout provides the greatest level routine, repetitive, and integral to the production process, a risk assessment should be performed. If the task performed poses an unacceptable risk, acceptable risk reduction methods should be implemented to reduce the risk to acceptable levels. PMID- 17424955 TI - ANSI Z358.1 compliance: check yourself out! PMID- 17424956 TI - Complying with OSHA's hexavalent chromium standards. PMID- 17424958 TI - Sounds good to me. PMID- 17424959 TI - Wearing a hard hat is only half the job. PMID- 17424960 TI - Is your equipment a silent hazard? Part II: Self-retracting lifelines. PMID- 17424961 TI - These problems aren't set in stone. PMID- 17424962 TI - Job-made guardrails: are they strong enough. PMID- 17424963 TI - Heroes on call. PMID- 17424964 TI - Remember the heat's on. PMID- 17424965 TI - [Image guided therapy]. PMID- 17424966 TI - [Seven cases of posterior cerebral artery aneurysms]. AB - In a period of ten years, since January 1996, we have encountered seven cases of posterior cerebral artery aneurysms (0.58% of surgically treated 1027 aneurysms in our hospital). Six of these cases were females (from 39 years old to 73 years old, average 54.5 years old). In five of the cases, neck clippings were performed, using the pterional approach for one P1 and one P2a aneurysm, the remaining three were clipping using the subtemporal approach for one P2a and two P2p aneurysms. In one case diagnosed as "unknown SAH" by typical findings of the CT scan and normal four-vessel study at the time of admission. The second vertebral angiography (two weeks later) revealed a P2a aneurysm. Such a case attracted our interest in the clinic, because many similar cases might have been diagnosed as "unknown SAH". In the case of a 42-year-old male with intracerebral hematoma, four-vessel study made a definite diagnosis as moyamoya disease and revealed a P2a aneurysm, which we cured by embolization of GDC for the preservation of anastomosis networks which had already been formed. Intravascular embolization is very effective, but sufficient studies are required before applying it in the cases of large and/or thrombosed aneurysms. We described surgical approaches about pterional, subtemporal and transchoroidal approaches and discussed some of the differences in the classification methods of Pia, Yasargil, Zear & Rhoton and Seoane. In particular, attention is necessary because we have confused Yasargil's classification with Zeal & Rhoton's classifications by using of the same notation, "Pn". PMID- 17424967 TI - [Deep brain stimulation of the posterior subthalamic area (Zi/Raprl) for intractable tremor]. AB - Tremor in the proximal arm muscle, trunk, or legs is often resistant to the standard stereotactic surgery of the thalamic ventrointermediate nucleus. We have performed deep brain stimulation (DBS) of the posterior subthalamic area for those intractable tremors. The white matter area between the red nucleus and the subthalamic nucleus was targeted on the T2-weighted MR-CT fused image. Inhibitory effect on the tremor was tested with macrostimulation. The somatosensory-evoked potential recorded through DBS contacts demonstrated characteristic biphasic pattern. Eight cases with severe essential tremor and 18 of tremor-dominant Parkinson disease were treated with unilateral DBS of the area including the zona incerta and the prelemniscal radiation (Zi/Raprl). Tremors including the proximal part have been well controlled for 24 months after the operation. The stimulation parameters have been almost stable during the follow-up period. There was no obvious adverse effect of stimulation. We conclude that Zi/Raprl-DBS is a safe and effective treatment on Parkinsonian and essential tremor. PMID- 17424968 TI - [Three cases of de novo aneurysms]. AB - We report 3 cases of de novo aneurysms that developed long after neck clipping of the initial aneurysms (range, 7 to 20 years, mean 12 years). Case 1 was a 58-year old female who had undergone clipping of a ruptured aneurysm 17 years previously. Ten years later, she suffered another subarachnoid hemorrhage due to rupture of a new aneurysm, for which neck clipping was performed. Another seven years later, she had a third subarachnoid hemorrhage, and angiography revealed a new aneurysm, for which neck clipping was performed. Case 2 and 3 were both 68-year-old females who had suffered subarachnoid hemorrhage with two aneurysms and had undergone neck clipping for them respectively, 20 years and 12 years previously. Each patient was admitted to our hospital with complaints of headache, diplopia, vertigo, etc., and newly formed aneurysms were detected by magnetic resonance angiography. Conventional angiography revealed three and one new aneurysms, respectively. Since case 1 was a special case of multiple aneurysms in which lesions appeared in series rather than in parallel, all three patients harbored multiple aneurysms. It is recommended that patients with multiple aneurysms, especially those after a long period postoperatively, undergo periodic examination on an outpatient basis to detect formation of de novo aneurysms by magnetic resonance angiography or 3D-CT angiography. PMID- 17424969 TI - [The surgical treatment of non-functioning pituitary adenomas in the ninth decade]. AB - There are no reports of surgical treatment for non-functioning pituitary adenomas in the 9th decade of life, and it is thought that these patients often select conservative treatment and observation, because of their worry about deterioration of their general condition due to invasive surgery and because hypopituitarism and other medical problems are often present. However, it is also true that there is necessity for considering surgical treatment for the pituitary adenomas even in the 9th decade when there is a complaint of visual disturbance. We carried out the palliative surgical removal of three non-functioning pituitary adenomas with visual disturbance in the 9th decade and report these surgical treatments and outcomes. The improvement of visual disturbance was obtained in all three cases. The only perioperative complication was pneumothorax caused by barotrauma. The palliative transsphenoidal surgical removal of non-functioning pituitary adenomas for the purpose of improvement of visual disturbance is a safe procedure in the 9th decade of life. PMID- 17424970 TI - [Usefullness of intrasacral fixation in an extremely unstable lumbosacral spine]. AB - Intrasacral fixation technique devised by Jackson is said to provide rigid lumbosacral fixation. We treated 3 cases of lumbosacral lesions using this technique in which lumbosacral segment had become extremely unstable during surgical intervention adding to the effect of original lesions. In all cases, surgeries were performed in 2 stages, intrasacral fixation and anterior stabilization. Case 1: A 52-year-old male was diagnosed fungal discitis and spondylitis at L4 and L5. X-ray showed destruction of the vertebral bodies. L2, L3 and sacrum were fixed posteriorly using the intrasacral fixation technique. One week after the first operation, L4 and L5 vertebral bodies were replaced by long fibula grafts through the extraperitoneal approach. Case 2: A 25-year-old female with cauda equina syndrome and abnormal body form diagnosed as having spondyloptosis in which the entire vertebral body of L5 had descended below the endplate of S1. MR imaging revealed marked canal stenosis at the S1 level. In the first surgery, L5 vertebral body was resected through the transperitoneal approach. After 1 week of bed rest, posterior segments of L5 were resected, L4 was affixed to the sacrum and anterior stabilization was achieved with 2 mesh cages and lumbosacral spine was fixed using the intrasacral fixation technique. Case 3: A 64-year-old female was diagnosed as having pyogenic discitis and osteomyelitis at the L5-S1 level. In spite of successful medical treatment for infection, low back pain continued. Radiologically, L5 vertebral body was shown to have collapsed and slipped anteriorly over the sacrum. L3, L4 and sacrum were fixed by intrasacral fixation. One week after the first operation, the L5/S1 disc and the suppurtive vertebral bodies were resected through the extraperitoneal approach and anterior stabilization was performed with iliac bone grafts. At follow-up for a minimum of 6 months, initial fixation was maintained in all 3 cases and bony fusion was obtained. The intrasacral fixation technique was considered to be effective for extremely unstable lumbosacral lesions. PMID- 17424971 TI - [A case of hydrocephalus in follow-up of post-traumatic subdural effusion]. AB - We report an interesting case of hydrocephalus following traumatic subdural effusion. A 50-year-old male was diagnosed as a traumatic subdural effusion. Three months later, he was transferred to our hospital again because of conscious disturbance and incontinence. Emergent CT showed characteristic hydrocephalus. The lateral ventricle and the third ventricle were remarkably enlarged. After Ventriculo-peritoneal shunt, the symptoms and radiographical findings were resolved. Both the compression of arachnoid villi around the superior saggital sinus and stenosis of the aqueduct by subdural effusion could be associated with the cause of hydrocephalus in this case. PMID- 17424972 TI - [Translational research for neurostimulation]. AB - The effects of electrical stimulation on the nervous system have been known for many years. Electrical stimulation has been employed as a treatment of functional diseases of the nervous system and has been mainly developed on the basis of deep brain stimulation (DBS), motor cortex stimulation (MCS), and spinal cord stimulation (SCS). Over the past two decades, DBS has replaced lesioning techniques as a treatment for movement disorders, and DBS has been shown to be safe and efficacious. The field of application for DBS is becoming wider, currently encompassing dystonias, epilepsy, and psychiatric disease. On the other hand, MCS has been employed as a therapy for intractable pain for more than ten years. Recently, this therapy has been used as a treatment for involuntary movements and motor palsy caused by stroke. Although the effects of electrical stimulation are distinct and the therapeutic benefits are well recognized, the exact mechanisms of these therapies have not yet been fully clarified. Animal experiments, either in vitro or in vivo, show complex patterns roughly suggesting inhibitory effects. More recent data from in vitro biological research studies show that electrical stimulation affects cellular functions, particularly protein synthesis, suggesting that it could alter synaptic transmission and reorganize neural networks. Research for the understanding of the effects of electrical stimulation at the neuronal level is still in progress, and knowledge from these research will eventually improve the effectiveness of this therapy. Moreover, these basic research and preliminary clinical studies will call for the re examination of the basic effects of electricity on living tissues because it is now clear that electrical stimulation has a wider application. In this review, we summarizes the history and rationale behind electrical neurostimulation therapy and the recent applications of this technique for various functional diseases of the nervous system. PMID- 17424973 TI - [Ethical and legal issues of regenerative medicine using human ES cells]. PMID- 17424974 TI - A long-acting calcium antagonist over one year did not improve BMIPP myocardial scintigraphic imagings in patients with pure coronary spastic angina. AB - BACKGROUND: Calcium antagonists (Ca) have been effective in reducing angina attacks in patients with variant angina. However, there are no reports regarding the effectiveness of Ca on myocardial fatty acid metabolic images in patients with pure coronary spastic angina (CSA). OBJECTIVES: This study sought to examine the correlation between myocardial fatty acid metabolic images and the medical treatment of Ca in patients with pure CSA. METHODS AND RESULTS: This study included 35 consecutive patients (28 men, mean age of 66 +/- 10 years) with angiographically confirmed coronary spasm and no fixed stenosis. Long-acting Ca was administered to all 35 patients. Isosorbide dinitrate /nicorandil/another Ca/beta-bloker were administered when chest pain was not controlled. Using an iodinated fatty acid analogue, 15-(p-[iodine-123]iodophenyl)-3 (R,S)methylpentadecanoic acid (BMIPP), myocardial scintigraphies with intravenous adenosine triphosphate infusion were performed before cardiac catheterization and 12 mo after medical therapy. According to the medical control states, these 35 patients were classified into 3 groups; response (disappearance of angina attacks, 12 pts, 60 +/- 11 years), partial response (angina attacks < 4/mo, 12 pts, 67 +/- 10 years), and no response to therapy (angina attacks > or = 4/mo, 11 pts, 71 +/- 6 years). Reduced BMIPP uptake was observed in 24 (69%) of 35 patients before the treatment. Reduced BMIPP uptake was also found in 18 patients (51%) after 12 mo. Normal BMIPP uptake after 12 mo therapy was observed in about half (response: 42%, partial response: 58%, no response: 45%) of patients among the 3 groups. There was no difference regarding the value of washout rate (WOR) (response; 10 +/- 7 (before), 14 +/- 8% (12 mo)), partial response; 11 +/- 7, 10 +/- 5%, no response; 13 +/- 9, 14 +/- 8%) among the 3 groups. The defect scores of BMIPP in the three groups were not different during at least one year medical therapy. No difference regarding the distribution of other medical therapies (angiotensin converting enzyme inhibitors/angiotensin receptor blockers/beta blockers/statins) was found. The administration of Ca and isosorbide dinitrate/nicorandil and 2 Ca was significantly higher in the poor than in the good control patients. CONCLUSIONS: Long-acting Ca over one year did not improve myocardial fatty acid metabolic images in patients with pure CSA. This may be related to silent ischemia. PMID- 17424975 TI - Clinical evaluation of the effect of attenuation correction technique on 18F fluoride PET images. AB - OBJECTIVE: The purpose of this study is to evaluate effect of attenuation correction technique on 18F-fluoride positron emission tomography (PET). METHODS: We performed PET scans after the injection of 185 MBq 18F-fluoride on 32 patients from October 20th, 2004 to April 13th, 2005. We calculated bone-to-muscle ratios for the images with and without attenuation correction. We placed regions of interest (ROIs) on normal bone accumulation in 22 patients. The exclusion criteria were bone metastasis, Paget's disease, and rheumatoid arthritis. Several regions were chosen for ROI placement: skull, cervical vertebra, mandible, scapula, thoracic vertebra, rib, humerus, lumbar vertebra, radius, ulna, pelvis, femoral head, femoral shaft, tibia, and fibula. The count ratios of normal bones to gluteus muscle were calculated as bone-to-muscle ratios. The count ratios of abnormal skeletal lesions to gluteus muscles were calculated as bone-to-muscle ratios, while the count ratios of abnormal skeletal lesions to normal bones were calculated as bone-to-bone ratios. RESULTS: PET images without attenuation correction showed significantly higher mean bone-to-muscle ratios than those with attenuation correction (p < 0.05) for all normal bones except the femoral head and lumbar vertebrae. For abnormal bones, bone-to-muscle ratios without attenuation correction were significantly higher than those with attenuation correction (p < 0.005). The same statistical significance was found for bone-to bone ratios (p < 0.005). CONCLUSIONS: The attenuation correction technique is not necessary to conduct the visual interpretation of 18F-fluoride PET images. The bone-to-muscle ratio analysis without attenuation correction may be of use to differentiate malignant from benign disease processes. PMID- 17424976 TI - Initial evaluation of dynamic human imaging using 18F-FRP170 as a new PET tracer for imaging hypoxia. AB - 18F-FRP170, 1-(2-fluoro-1-[hydroxymethyl]ethoxy)methyl-2-nitroimidazole, is a new hypoxia imaging agent for positron emission tomography. This compound was synthesized by 18F-labeling of RP170, which was developed as a new hydrophilic 2 nitroimidazole analog. In the present study, we analyzed dynamic whole-body imaging in healthy volunteers and dynamic tumor imaging in three patients with lung cancer. METHODS: Four healthy male volunteers and three lung cancer patients were enrolled in this study. Volunteers underwent dynamic whole-body scans just after injection of 18F-FRP170 for about 90 min, while the lung cancer patients underwent dynamic tumor imaging for about 60 or 120 min. Data are expressed as standardized uptake values (SUV). Regions of interest were placed over images of each organ or tumor to generate time-SUV curves. RESULTS: The series of dynamic whole-body scans showed rapid elimination of 18F-FRP170 from the kidneys following elimination from the liver. Very low physiological uptake was observed above the diaphragm. 18F-FRP170 uptake in the lung cancer lesion could be visualized clearly from early after injection. The changes of tumor SUV, tumor/blood ratio, or tumor/muscle ratio about 30 min after injection or later were small. CONCLUSIONS: Dynamic imaging using 18F-FRP170 demonstrated rapid elimination from the kidney, suggesting the high hydrophilicity of this imaging agent. The background activity above the diaphragm was very low, and patients with lung cancer showed clear tumor uptake of 18F-FRP170 early after injection. PMID- 17424977 TI - Crossed cerebellar diaschisis: a positron emission tomography study with L [methyl-11C]methionine and 2-deoxy-2-[18F]fluoro-D-glucose. AB - OBJECTIVE: Crossed cerebellar diaschisis (CCD) is defined as a depression of blood flow and oxidative metabolism of glucose in the cerebellum contralateral to a supratentorial brain lesion, as detected with positron emission tomography (PET) and single photon emission computed tomography. We examined whether L [methyl-11C]methionine (MET) uptake is affected in CCD. METHODS: In 12 patients with a unilateral supratentorial brain tumor, we evaluated the uptake of 2-deoxy 2-[18F]fluoro-D-glucose (FDG) and MET in the cerebellar hemispheres by means of PET. Asymmetry index (AI) was defined as a difference in the average count between the ipsilateral and contralateral cerebellar hemispheres divided by the average count in both cerebellar hemispheres. Patients with AI of FDG PET more than 0.1 and those with AI equal to 0.1 or less than 0.1 were classified as CCD positive and CCD-negative, respectively. RESULTS: Six patients were CCD-positive and others were CCD-negative in the FDG PET study. Between CCD-positive and CCD negative patients, mean AI of MET was not significantly different (0.017 +/- 0.023 and 0.014 +/- 0.039, respectively). CONCLUSIONS: Different from glucose metabolism, cerebellar MET uptake was not affected in CCD. The present study may indicate that cerebellar MET uptake is independent of suppression of cerebellar neuronal activity. PMID- 17424978 TI - Global and regional evaluation of systolic and diastolic left ventricular temporal parameters using a novel program for ECG-gated myocardial perfusion SPECT--validation by comparison with gated equilibrium radionuclide angiography and speckle-tracking radial strain from echocardiography. AB - BACKGROUND: A newly developed program, named cardioGRAF, enabled the evaluation of left ventricular (LV) systolic and diastolic temporal parameters for the estimation of heart failure using ECG-gated myocardial perfusion SPECT (GMPS). OBJECTIVE: The feasibility of those global (g-) and regional (r-) parameters was validated to compare with gated equilibrium radionuclide angiography (ERNA) and speckle-tracking radial strain (STS) from echocardiography. METHODS: Thirty-three patients were studied using GMPS and ERNA (n=11) or GMPS and STS (n=22). The following g- or r-parameters obtained by cardioGRAF and ERNA or STS were compared: time to end systole (TES), time from end systole to peak filling rate (TPF1), time from 0 to peak filling rate (TPF2), time to peak radial strain (TPS), time from peak strain to peak negative strain rate (TP-SR1), and time from 0 to peak negative strain rate (TP-SR2). RESULTS: All g-parameters were successfully obtained by cardioGRAF and ERNA. The results demonstrated good correlations (g-TES: r = 0.79, p < 0.005; g-TPF1: r = 0.75, p < 0.02; TPF2: r = 0.83, p < 0.005). The differences were 11.9 +/- 31.8 ms in g-TES, 19.9 +/- 65.4 ms in g-TPF1, and 37.7 +/- 67.4 ms in g-TPF2. All r-parameters were successfully obtained by cardioGRAF. Eight patients and 12 segments were excluded because of the inadequate quality of routine echocardiography for STS analysis. However, r parameters obtained by cardioGRAF were significantly correlated with those of STS (r-TES and r-TPS: r = 0.61, p = 1 x 10(-8); r-TPF1 and r-TP-SR1: r = 0.69, p = 3 x 10(-11); r-TPF2 and r-TP-SR2: r = 0.76, p = 2 x 10(-15)). The differences were 22.1 +/- 38.2 ms between r-TES and r-TPS, 7.0 +/- 123.4 ms between r-TPF1 and r TP-SR1, and 38.1 +/- 111.5 ms between r-TPF2 and r-TP-SR2. CONCLUSION: The feasibility of evaluating systolic and diastolic temporal parameters by a new program was validated. This program has the potential to evaluate both diastolic and systolic heterogeneous wall motions which express dyssynchrony in heart failure. PMID- 17424979 TI - Advantages of upright position imaging with medium-energy collimator for sentinel node lymphoscintigraphy in breast cancer patients. AB - OBJECTIVE: To evaluate the advantage of upright position imaging with a medium energy collimator for the detection of sentinel lymph node (SLN). METHODS: Thirty four patients with operable breast cancer underwent sentinel node lymphoscintigraphy with 99mTc-tin colloid. Images were obtained in 5 different positions and paired images from the same patient were compared using side-by side interpretation. Images were compared in 3 groups: group 1 (anterior view); supine (SAV) vs. upright (UAV), group 2 (oblique view); supine (SOV) vs. upright (UOV), and group 3 (oblique view); modified supine (MOV) vs. UOV. Image quality was evaluated using a 3-grade scale of clear, faint, and equivocal depiction, and correlated to 3 parameters: distance from injection site to lymph node (hot node), counts in hot node, and image contrast. Parameters in group 1 were compared by classifying the primary tumor site into 4 subregions. RESULTS: Image quality in all 3 groups was more enhanced on the image obtained in the upright position than that in the supine position. Obtaining images in an upright position increased the mean distances by 1.5-3.2 cm, and mean contrasts were significantly increased by 0.13-0.31 (p < 0.05). It was shown that image quality was more greatly affected by image contrast than by counts in the hot node. Image contrast of 0.5 seemed an appropriate threshold level for detection of the hot node. On comparison of tumor sites, the upper outer quadrant (C) region of the 4 subregions demonstrated greater contrast enhancement on upright position images. CONCLUSION: Clinical images obtained in an upright position with a medium-energy collimator were superior to those obtained in a supine position. Use of this procedure is recommended to enhance lymph node detection on sentinel node lymphoscintigraphy. PMID- 17424980 TI - Diagnosis of maxillofacial tumor with L-3-[18f]-fluoro-alpha-methyltyrosine (FMT) PET: a comparative study with FDG-PET. AB - OBJECTIVES: To compare L-3-[18F]-fluoro-a-methyltyrosine (FMT)-positron emission tomography (PET) and 2-[18F]-fluoro-2-deoxy-D-glucose (FDG)-PET in the differential diagnosis of maxillofacial tumors. METHODS: This study included 36 patients (16 males, 20 females; 31-90 years old) with untreated malignant tumors (34 squamous cell carcinoma, one mucoepidermoid carcinoma, one rhabdomyosarcoma) and seven patients (five males, two females; 32-81 years old) with benign lesions. In all patients, both FMT-PET and FDG-PET were performed within two weeks before biopsy or treatment of the lesions. To evaluate the diagnostic usefulness of FMT-PET and FDG-PET, visual interpretation and semiquantitative analysis were performed. PET images were rated according to the contrast of tumor uptake as compared with background, and were statistically analyzed. As a semiquantitative analysis, standardized uptake values (SUV) of the primary tumors were measured, and the SUV data were analyzed using receiver operating characteristic (ROC) curves. RESULTS: The mean SUV of the malignant lesions were significantly higher than those of the benign lesions in both FMT-PET (2.62 +/- 1.58 vs. 1.20 +/- 0.30, p < 0.01) and FDG-PET (9.17 +/- 5.06 vs. 3.14 +/- 1.34, p < 0.01). A positive correlation (r = 0.567, p < 0.0001, n = 46) was noted between FMT and FDG. ROC analysis revealed that there was no statistically significant difference in SUVs between FMT and FDG for differentiating malignant tumors. In 27 of 36 patients, FMT-PET had better contrast of malignant tumor visualization to the surrounding normal structures by visual assessment (p < 0.005, binomial proportion test). CONCLUSIONS: Differential diagnosis of FMT-PET based on the uptake in maxillofacial tumors is equivalent to FDG-PET. However, the contrast of FMT uptake between maxillofacial tumors and the surrounding normal structures is higher than that of FDG, indicating the possibility of accurate diagnosis of maxillofacial tumors by FMT-PET. PMID- 17424981 TI - [Diagnostic of cognitive dysfunctions in multiple sclerosis]. AB - Cognitive dysfunctions are frequent symptoms in multiple sclerosis (MS). Up to 65% of the patients suffer from cognitive dysfunctions. Especially memory, attention and executive functions are impaired. These problems strongly affect the patients' ability to work and their quality of life. A differentiating diagnostic effort is necessary to control fatigue and depression. Screening tools alone cannot provide a detailed description of all cognitive domains. Therefore, an elaborated neuropsychological diagnostic is necessary. This report provides a description of cognitive functions and its diagnostic opportunities, especially in MS patients. After displaying aspects of differential diagnostics, a recommendation for a diagnostic work schedule is given. PMID- 17424982 TI - [Assurance-associated aspects of head and neck carcinomas: disposition, exposition, prognosis]. AB - After a brief introduction into the basics of head and neck oncology, the current review focusses on potential predictive markers and predisposing factors for these kinds of cancers. Furthermore, prognosis and degree of disability of cancer patients are commented on. With a survival rate of approximately 50%, the prognosis of head and neck carcinomas seems rather good, but it is in fact unsatisfactory. Since the treatment of advanced head and neck cancers is often accompanied by profound functional (articulation, phonation, respiration, and deglutition) and aesthetic impairment, it repeatedly leads to a high degree of disability and occupational invalidity. Furthermore, special limitations of head and neck cancer patients are discussed as well as the consequences they may have on the qualification for certain jobs. PMID- 17424983 TI - [The complex regional pain syndrome (CRPS) as differentiation from psychogenic disturbances]. AB - In the course of recent years, after the conception of dystrophic disturbances of the extremities had been reassessed, the CRPS has been making increasing demands on medical treatment and assessment in all insurance classes. There is common agreement that the syndrome can also be triggered by minor trauma. A decisive factor for the expert opinion is first of all the presentation of definite proof that there is actually a dystrophic disturbance present on the affected extremities. The patients concerned frequently suffer from a co-morbidity with accompanying psychological disturbances. For a lot of patients, primary organic effects caused by the injury can be excluded, whereas psychogenically determined symptoms can be found. This leads to significant consequences for the medical assessment under the insurance aspect in the individual case. The differential diagnosis may turn out to be difficult and in individual cases will require the use of the entire available technical diagnostics equipment and the involvement of psychological expert opinions. PMID- 17424984 TI - [Hannover-Polytrauma-Longterm-Study HPLS]. AB - From the period of September 2000 to March 2006 Hannover Reinsurance and the Department of Traumatology of the Hanover Medical School conducted a retrospective observational study of the long-term outcome of patients after polytrauma. The follow-up period was on average 17 +/- 5 years. The goal of the study was to determine the social, financial, medical and psychological long-term outcome after a severe accident. Of the 1560 enrolled patients 519 patients died in the observed period, 397 patients were followed up but could not be examined, 637 patients (female n = 158, male n = 479) were examined and answered detailed questionnaires concerning their social, financial medical and psychological situation. 6 patients did not fullfil the input criteria and one patient had two polytraumata. The average age at the time of accident was 26.5 years (+/- 12.3). RESULTS: Financial losses were observed in 41.1% of all males and 44.4% of females. 21.7% of all male (female 18.4%) had no financial protection at all. Due to the accident 30.1% of all men were unemployed and 19.8% permanently disabled (women 27.4% and 27.6% respectively). Psychological treatment almost doubled after discharge from rehabilitation centres. Even 20 years after the accident extra-mortality was substantially increased by 79%. CONCLUSION: Ten to twenty years after polytrauma significant financial, social and medical impairments are still present. PMID- 17424985 TI - [Extent to which a claimant's loss-mitigation and cooperation obligations towards the insurer are enforceable--requirements for an expert medical opinion]. AB - A policyholder claiming an annuity benefit on the grounds that, he is no longer able to pursue his occupation must meet certain obligations vis-a-vis the insurer, e.g. duty to mitigate the loss, duty to cooperate with the insurer. These obligations are often curtailed by the claimants' rights. Physicians and providers of expert medical opinions frequently do not share this opinion. Where medical and legal considerations intermingle and, potentially, conflict, there is great uncertainty as to whether, and to what extent, an insurer may require a claimant to undergo medical treatment. Before deciding on whether or not to grant an annuity, the insurer generally calls in medical experts to assess the policyholder's actual degree of disability and offer an opinion on whether and, if so, by what medical means that disability might be mitigated or remedied. Expert medical opinions are often decisive in such cases. The present article discusses the extent to which a claimant's loss-mitigation and cooperation obligations towards the insurer are enforceable; in this context, a number of court decisions are quoted. In addition, the authors define the requirements that an expert medical opinion must meet. PMID- 17424986 TI - [A new scale for measuring return-to-work motivation of mentally ill employees]. AB - A new scale "motivation for return to work" has been constructed to measure depressive patients' motivation to start working again in a stepwise process. The scale showed in 46 patients of a first case management (CM) sample with depressive employees a good correlation with the final social status of the CM. Only the motivated patients were successful returning to work and could be, separated clearly from the most demotivated one. Second, the scale correlated with the duration of sick leave and third showed an inverse correlation with the complete time of CM, suggesting that a successful stepwise return to work requires time. These first results need further examination. PMID- 17424987 TI - [Macroeconomic pitfalls of the national health program for the resources of private health insurance]. PMID- 17424988 TI - [Expensive health resort treatment in beautiful Switzerland]. PMID- 17424989 TI - [Significance of occupational old age pensions for clinics]. PMID- 17424990 TI - [New targets for the treatment of malaria]. PMID- 17424991 TI - [Trivirus--the multivirus killer]. PMID- 17424992 TI - [Green tea against AIDS?]. PMID- 17424993 TI - [The history of treatment lipid metabolism disorders]. PMID- 17424994 TI - [Lipid and lipoprotein metabolism]. PMID- 17424995 TI - [Molecular pharmacology and medicinal chemistry of fibrate]. PMID- 17424996 TI - [Fibrate in lipid metabolism disorders]. PMID- 17424997 TI - [Nicotinic acid in lipid metabolism disorders]. PMID- 17424998 TI - [The use of fish oil in lipid metabolism disorders]. PMID- 17424999 TI - [Plant omega 3- and omega 6-fatty acids]. PMID- 17425000 TI - [New targets and drugs for treatment of lipid metabolism disorders]. PMID- 17425001 TI - [Human skull development and voice disorders]. AB - The hominisation of the skull comes with the bipedic posture, due to a network of muscular and aponevrotic forces applied to the cranio-facial skeleton. A brief sight of the morphogenetic origine and issues of these forces help to understand more clearly the postural statement of the larynx, his functions, and his many extrinsic biomechanical bounds; then further his most frequently dysfunctions. The larynx is surrounded by several effective systems of protection: active, activo-passive, passive. The architectural features of the components of the laryngeal system allows us to consider the laryngeal function as an auto-balanced system. All the forces engaged are auto-balanced in a continuum of tension. This lead us to the concept of tensegrity system, neologism coming from tensional integrity described by Buckminster Fuller. The laryngeal employement by extrinsic system is pathological in case of chronicity. Any osteopathic treatment, which aims to restore the losses of laryngeal mobility, has to release first the peripherical structures involved in the laryngeal defense, before normalising the larynx itself Finally, the larynx recovers his functions in a tensegrity system. PMID- 17425002 TI - [Vocal forcing and posture: experimental studies on normal subject]. AB - One of the well-known characteristics of vocal forcing is postural with an increase in the antero-posterior movements of the trunk and head during phonation. OBJECTIVE: we conceived different physiological experiments on normal subjects to explore in an objective way these movements. MATERIALS AND RESULTS: A series of experiments using a platform of posturography confirmed that there is an increase in the tensions in the muscles implied in the posture when the subject forces his voice because of an ambient noise. This increase is characterized by the index VCOP rms (variance of the displacement of the center of pressure in upright position) which passes from 13.19 in normal voice to 18.63 in forced voice. A complementary study was carried out with an equipment of analysis of the movements (ELITE). CONCLUSION: We could, thus, confirm the existence of the contemporary antero-posterior movements of vocal forcing. The discussion concerns the application perspectives of these experimental knowledge in the clinical field of the dysfunctional dysphonia. PMID- 17425003 TI - [Vocal effectiveness in speech and singing: acoustical, physiological and perceptive aspects. applications in speech therapy]. AB - OBJECTIVE: What is vocal effectiveness in lyrical singing in comparison to speech? Our study tries to answer this question, using vocal efficiency and spectral vocal effectiveness. MATERIAL AND METHODS: Vocal efficiency was mesured for a trained and untrained subject. RESULTS: According to these invasive measures, it appears that the trained singer uses her larynx less efficiently. Efficiency of the larynx in terms of energy then appears to be secondary to the desired voice quality. The acoustic measures of spectral vocal effectiveness of vowels and sentences, spoken and sung by 23 singers, reveal two complementary markers: The "singing power ratio" and the difference in amplitude between the singing formant and the spectral minimum that follows it. Magnetic resonance imaging and simulations of [a], [i] and [o] spoken and sung show laryngeal lowering and the role of the piriform sinuses as the physiological foundations of spectral vocal effectiveness, perceptively related to carrying power. CONCLUSION: These scientifical aspects allow applications in voice therapy, such as physiological and perceptual foundations allowing patients to recuperate voice carrying power with or without background noise. PMID- 17425004 TI - [Epidermoid carcinoma of piriform sinus after chemo-radiotherapy: acoustic evaluation and voice handicap]. AB - OBJECTIVE: The aim of this prospective investigation was to analyse the voice and the vocal handicap for patients who received concomitant chemoradiation therapy in piriform sinus epidermoid carcinoma (T2, T3). MATERIAL AND METHODS: This study included two males populations: Twelve patients aged from 48 to 71 years old were evaluated between 6 and 30 months after the end of chemo-radiotherapy and ten normal subjects, matched in age. The evaluation consisted in firstly a vocal recording (vowel and text) allowing an objective acoustic analysis (temporal and frequential parameters) and a perceptive evaluation of the voice quality by means of GRBAS scale. Secondly an evaluation of the quality of life by means of the voice handicap index (VHI). RESULTS: The acoustic parameters of instability for the fundamental frequency (FO) and the maximum phonation time of the patients were statistically different (p <0.05) related to the normal subjects. On the other hand, the FO (patients - 130 Hz, subjects = 121 Hz) as well as the speech rate (patients = 142 words/mn, subject = 148 words/mn) were not statistically different. The perspective indexes showed a deterioration of the vocal quality while the handicap expressed was minor. CONCLUSION: This preliminary study showed the interest of evaluations of the vocal function and the quality of life after chemo-radiotherapy for piriform sinus carcinomas allowing patients to conserve a phonation and eventually the propose a speech therapy. PMID- 17425005 TI - [Quality of life after oropharyngectomy]. AB - OBJECTIVE: The purpose of this study was to assess direct or non-direct links between functional abilities of the patient evaluated objectively and his quality of life after oropharyngectomy, a surgery capable of engendering bad functional outcomes. METHODS: Praxies and speech were examined in 16 patients at 3, 6, 9 or 12 months after surgery and 2 self-evaluation questionnaires were proposed: One about health related quality of life of cancer patients (EORTC QLQ-C30 and H&N35) and, the other about oral communication handicap (VHI). RESULTS AND CONCLUSION: A correlation between quality of life and objective functional abilities can not always be found. Self-evaluation questionnaires might be a complementary approach in order to guide management in patient care after oropharyngectomy. PMID- 17425006 TI - [Validation of a self-assessment questionnaire for swallowing disorders (Deglutition Handicap Index)]. AB - OBJECTIVES: The aim of this study is the psychometric validation of a self assessment questionnaire about swallowing disorders among adults, insisting on the validity of its content. MATERIAL AND METHOD: Created on the model of the "Voice Handicap Index", this questionnaire is composed of 30 statements on deglutition related aspects in daily life (5 point-rating scale). It is subdivided in three domains of 10 items: physical (S) (symptoms related to swallowing), functional (F) (nutritional and respiratory consequences) and emotional (E) (psychosocial consequences). Our first goal was to verify wether the different domains of the questionnaire were well related to what we wished to quantify or not. Therefore, regarding the domain S we compared the physiopathological mechanisms assessed by videofluoroscopy examination, for the domain F its results compared to the nutritional status and for domain E a scale of quality of life. RESULTS: 96 patients with dysphagia and 53 normal subjects were included in the study. The internal consistency reliability was correct (Cronbach's alpha= 0.9), For the domain S the difference between the normal subjects and the patients was statically significant. Our hypothesis regarding the domains were confirmed but not specifically. The domain S is related to the physiopathological mechanisms but also to the degree of severity. There are some correlations between the domain F and the nutritional status but also the results of the videofluoroscopy and the scale of quality of life. The domain E presents a fair correlation with the quality of life but it's related too much to physiopathological mechanisms, the duration of meal and the severity. As a conclusion, the results of this study allows to resume the validation without modification in the questionnaire. We are thinking, because of its similarity with the "Voice Handicap Index", to name it the "Deglutition Handicap Index". PMID- 17425007 TI - [The rehabilitation of the tongue in hypoglossal-facial anastomosis]. AB - End to end hypoglossal-facial nerve anastomosis (tt HFA) is a traditional technique for rehabilitation of facial palsy. The sacrifice of the hypoglossal nerve generates a paralysis and an atrophy of the tongue which is thought to lead to speech, chewing and swallowing disorders. In a previous study, we demonstrated that tt HFA does not lead to speech disorders (Gatignol et al 2003). OBJECTIVES: In this work, we were interested in the functional consequences of the lingual atrophy and in the possibilities of rehabilitation with early therapy. MATERIAL AND METHODS: Nine patients were distributed in two groups, in one patients received a specific and early lingual rehabilitation in the other no specific treatment. These two groups (paired with control subjects) were subjected to a series of tests studying the motor function, the articulation (using palatograms). RESULTS: This study highlights the interest of early rehabilitation of the tongue in the first post-operative days. Early rehabilitation was associated with a reduction in lingual atrophy, an improvement in motility of the tongue thus generating a better management of saliva and stagnant food in the oral vestibule on the paralysed side. PMID- 17425008 TI - [Training strategies of the spontaneous voice in the reeducation of the speech voice]. AB - Rehabilitation cannot by definition be spontaneous, it must approach it by exercises which it proposes. It is the objective of the various strategies exposed here. The strategic points of the rehabilitation of the vocal spontaneousness of the spoken voice are: The articulatory precession of the vowels related to the pre-phonatory inspiratory impulse, the reduction of the dimension of a voluntary movement adapted to an automatic movement, the points of rocking of the singing voice towards the spoken voice, the slip of vocalization towards the prosody, the projected prosodic counting, the simple expressions whose dynamic model is known socially of all, the reading with under text of transcriptions of interview of famous authors. It is in this step and this progression that one can ensure oneself of a vocal recovery reaching the spontaneousness which the word needs to be and remain alive. PMID- 17425009 TI - [Endoscopic medialization with Vox implant: our experience]. AB - OBJECTIVE: The purpose of this work is to analyze our preliminary results on the endoscopic vocal cords medialization by Bioplastique. MATERIAL AND METHOD: This was a retrospective study on 39 cases of unilateral laryngeal paralysis, treated with the injection of Vox-implant between May 2004 and August 2006. The evolution of the swallowing disorders was followed by video fluoroscopy. The phoniatric evaluation was based on an auto evaluation questionnaire: Voice Handicap Index 10 (VHI-10). RESULTS: Of the 19 patients who presented with swallowing disorder, 18 gave a normal video fluoroscopy test. In only one case, a second injection, one year later was necessary to completely cure the patient from fluid aspirations. Thirty two patients were satisfied with the post-operative vocal results. The Wilcoxon matched pairs test gave a statistically significant difference between pre and post-operative VHI-10 (p< 0,001). The voice handicap improved approximately 53%. CONCLUSION: Bioplastic Vox-lmplant profits from a simple injection technique, it is well tolerated and gives a good voice quality. It allows moreover a fast and simple resolution of the problem, can be repeated, and in case of failure or insufficiency a thyroplasty remains possible. PMID- 17425010 TI - [Laryngeal pemphigus]. AB - The pemphigus vulgaris is a serious bullous disease of the adult. The pharyngolaryngeal localization has rarely been described. The oral and genital localizations are often inaugural. We report the case of a 45 year old female who presented a laryngeal pemphigus which evolved during several years before its cure. The bullous dermatosis are serious affections capable of being stressfull to life. Their diagnosis and treatment must be done quickly. The discovery of painful pharyngolaryngeal lesions, erosive or bullous, must lead to a biopsy with immunofluorescence histological examination in order to diagnose the pemphigus vulgaris and conduct quickly the treatment. PMID- 17425011 TI - [The "intra-cordal polyp": a clinical alternative to recognize?]. AB - The vocal cord polyp is an inflammatory false tumour of the larynx. It is characterized mainly by the specific existence of fibrinous exsudats organized in loose network or mounds, surrounded by newly formed vascular slits. The epithelium covering the polyp is usually more or less impaired. CLINICAL CASES: The authors report two adult patients among whom the initial presentation in video-stroboscopy was that of an intracordal lesion of a cystic type, however; with unusual characteristics (purplish color, angiomatous aspect). The surgery consisted of a cordotomy, the surface epithelium appearing normal. After dissection, the two lesions seemed to be presenting the typical aspect of fibrinoid mound of a polyp, observation confirmed by the anatomo-pathological study. In these cases, the resection was finally carried out with no mucous loss. CONCLUSION: The particular etiopathogeny of these lesions is been discussed. The treatment, medical and of rehabilitation could achieve a partial recovery. In the event of surgery, an access to the lesion through a cordotomy and not directly by removal with microcissors is advised. PMID- 17425012 TI - [Lateral fixation of the vocal fold (Lichtenberger's technique): interest in the bilateral laryngeal immobilities]. AB - In patients presenting a bilateral laryngeal immobility, the potential reversibility of certain cases, the refusal or the bad tolerance of long term tracheotomy bring up the question of the choice of the surgical technique if it is indicated. OBJECTIVES: To report our experience with the technique of lateralization of the paralyzed vocal fold (arytenoidopexy) suggested by Lichtenberger. CLINICAL CASES: After having described the technique, we report 5 cases (3 pos-thyroidectomy, I of central origine, 1 post-burn). From 1 to 12 months after surgery 2 patients were fully satisfied, a patient (central origin) recovered spontaneously at the end of a month and the 2 last had a partial result. Only one patient required several surgical gestures. CONCLUSION: The Lichtenberger's technique combines theoretical reversibility and conservation of a functional glottic plan. It avoids tracheotomy. This approach can according to us validly replace the traditional techniques, medium or long term tracheotomy , or endoscopic arythenoid or posterior vocal fold resection. PMID- 17425013 TI - Snapshot at the end of 2006. PMID- 17425014 TI - Revenue cycle management, Part II. AB - The proper management of your revenue cycle requires the application of "best practices" and the continual monitoring and measuring of the entire cycle. The correct technology will enable you to gain the insight and efficiencies needed in the ever-changing healthcare economy. The revenue cycle is a process that begins when you negotiate payor contracts, set fees, and schedule appointments and continues until claims are paid in full. Every single step in the cycle carries equal importance. Monitoring all phases and a commitment to continually communicating the results will allow you to achieve unparalleled success. In part I of this article, we explored the importance of contracting, scheduling, and case management as well as coding and clinical documentation. We will now take a closer look at the benefits charge capture, claim submission, payment posting, accounts receivable follow-up, and reporting can mean to your practice. PMID- 17425015 TI - Prepare your practice for pandemic flu. AB - Highly virulent H5N1 Asian bird flu is poised to arrive in the United States. If the avian flu virus mutates, becoming transmissible from person to person, widespread disease is probable. Presuming World Health Organization predictions hold, patient demand may overwhelm medical facilities making pandemic influenza preparation and coordination essential. This article provides a checklist to use to organize your pandemic flu-planning efforts. Physician offices and other healthcare facilities must stay flexible in their response to a possible influenza pandemic. Prepare by planning in advance for the worst-case scenario. Revise your plan as needed to keep abreast of future developments. PMID- 17425016 TI - Netiquette: e-mail for group practices. AB - E-mail is a relatively new tool in business and communication, and it has unique nuances. Effective use of e-mail and its features can enhance performance, interaction, even morale. Misuse of e-mail can be a legal liability and damage relationships within your practice. This article provides a guide to optimizing the use of e-mail within your practice. PMID- 17425017 TI - Keeping time: avoiding the top not-quite-ten mistakes in employee overtime. AB - In addition to many other compliance areas a physician practice administrator must oversee, attention to employment law issues can minimize risk, as well as improve employee relations and morale. Wage/hour compliance should be at the top of an administrator's employment law to-do list. This article reviews the most common mistakes practices make when dealing with employee overtime issues. PMID- 17425018 TI - Record retention: what to keep, what to throw out. PMID- 17425019 TI - Cool, calm, and collected. AB - Selecting a collection agency--or evaluating the one you have--can improve bottom line results while maintaining your professional image. This article provides criteria for choosing a prospective collection agency and evaluating the strategies a collection agency employs, and discusses what a client medical practice should expect in analyses and support. PMID- 17425020 TI - Computer briefs: open science: the coming paradigm shift. PMID- 17425021 TI - Physician mentoring and evaluation. AB - Maintaining a cohesive medical group requires more than partners who get along with one another. Physicians must share the same values and be willing to give (and graciously receive) honest feedback on issues such as quality of care, technical competence, patient- and staff relations, behavior, work ethic, and productivity. This article shows group leaders how to start this process by mentoring new physicians and how to then extend the process to include all physicians in the group. Medical practices that have evaluation systems in place enjoy benefits that include better communication, accountability, increased retention rates, and a more unified group. Many physician groups avoid the evaluation process because they are not comfortable "judging" their peers, they don't know how to approach the process, or they don't want to invest the time. This article presents alternative approaches to establishing a mentoring and evaluation process, shows group leaders how to identify which is right for them, and provides do's and don'ts for a smooth implementation ofthe process. PMID- 17425022 TI - Top seven hospital billing questions. PMID- 17425023 TI - Broadbanding a surgical academic practice salary: a possible score. AB - How can we look at productivity in academic centers when surgeons perform different procedures that pay differently? Another dilemma is how they are compensated for teaching medical students, residents, and other surgeons. We also have to compensate them for the types and difficulty of the procedures. We can view this problem as either "making the pie bigger" or "dividing the pie better. "First, we should focus on how to "divide the pie. "Regardless of the "pie size," the issue of allocation for remuneration purposes is an important issue. "Dividing" the pie is an internal exercise, whereas making the pie "bigger" involves additional internal and external factors. In this paper, we address the issue of dividing the pie in a measurable way. We also address how to score each activity so that bonuses or compensation can be calculated without the "more" productive surgeons effectively subsidizing the "less" productive, a situation that is often detrimental to organizational success. Academic surgeons are very important for teaching new surgeons and medical students; therefore, they should be remunerated adequately. Pay schemes may be developed to improve the retention of highly productive surgeons in the academic environment. PMID- 17425024 TI - Mutual expectation compacts: a means to link practice culture and vision. AB - We describe the development of a mutual expectation compact in an academic department of surgery as a means of reinforcing the department's vision document and promoting cultural change. The compact makes explicit those implicit agreements that exist between a physician and his or her practice. It strengthens the relationship by avoiding the misunderstandings that can arise when agreements are implicit. PMID- 17425025 TI - How the medical practice employee can get more from continuing education programs. AB - Continuing education can be a win-win situation for the medical practice employee and for the practice. However, in order education programs must become informed consumers of such programs. They must know how to select the right educational programs for their needs and maximize their own participation. Employees who attend continuing education programs without preparation may not get the full benefit from their experiences. This article suggests benchmarks to help determine whether a continuing education program is worthwhile and offers advice for calculating the actual cost of any continuing education program. It provides a how-to checklist for medical practice employees so they know how to get the most out of their continuing education experience before, during, and after the program. This article also suggests using a study partner system to double educational efforts among employees and offers 10 practical tips for taking and using notes at a continuing education program. Finally, this article outlines the benefits of becoming a regular student and offers three practical tips for maximizing the employee's exhibit hall experience. PMID- 17425026 TI - Trends in the consumer price index. PMID- 17425027 TI - Physician management and leadership education at the Cleveland Clinic Foundation: program impact and experience over 14 years. AB - As the challenges of leading in healthcare become more complex, healthcare institutions have increasingly emphasized the importance of leadership training for physicians. Several institutions have offered in-house training programs. This paper describes the 14-year experience and evolution of physician leadership development training at the Cleveland Clinic. We describe the curriculum, organization, and goals of the Leading in Health Care course, which is offered to high-potential physician leaders. As a metric of the success of this physician leadership effort, we report the number and types of business plans developed in the course that have been either implemented at the Cleveland Clinic or have directly affected plans for program implementation. PMID- 17425028 TI - Get personal with customer service. PMID- 17425029 TI - Light-duty jobs and the Americans with Disabilities Act. PMID- 17425030 TI - Hospital financial health: increasing staff productivity. PMID- 17425031 TI - Hypoxic preconditioning prevents development of post-stress depressions in rats. PMID- 17425032 TI - Effect of derivatives of docosahexaenoic acid on AMPA receptors in Purkinje neurons. PMID- 17425033 TI - Theoretical evaluation of cumulative risk of altitude decompression sickness. PMID- 17425034 TI - Activators of Epac proteins induce relaxation of isolated rat aorta. PMID- 17425035 TI - High-protein diet during lactation changes functioning of enzyme systems of the small and large intestines of progeny in adultness. PMID- 17425036 TI - Immunomodulating effect of elicitors and suppressors upon infection of potato with Globodera rostochiensis (Wollenweber, 1923) Beherens, 1975. PMID- 17425037 TI - Some specific features of nitrogen fixation in the digestive tract of the European beaver (Castor fiber). PMID- 17425038 TI - Studies on bottom communities in the Green-Fjord, Svalbard. PMID- 17425039 TI - Tadpoles of the grass frog (Rana temporaria) and moor frog (Rana arvalis) avoid zones with elevated ammonia concentration. PMID- 17425040 TI - Octopamine regulates the 20-hydroxyecdysone level in Drosophila females. PMID- 17425041 TI - Changes in the external morphology of shells of freshwater bivalves of the superfamily Pisidioidea depending on the geographic location of their habitats. PMID- 17425042 TI - The diversity of Y-chromosome lineages in indigenous population of South Siberia. PMID- 17425043 TI - Effect of helminthic infestation on the biochemical parameters of gulls of the genus Larus in the Barents Sea region. PMID- 17425044 TI - Involvement of general transcriptional factors in the regulation of transcription of the hsp70 gene in vivo. PMID- 17425045 TI - Plant adaptation to salinity reduces copper toxicity. PMID- 17425046 TI - Generation of superoxide radical and hydrogen peroxide in insect hemolymph in the course of immune response. PMID- 17425047 TI - Relationships between plants and enterobacteria that are pathogenic for humans. PMID- 17425048 TI - Translational dynamics of water in the cytoplasm of parenchymal cells of Malus domestica fruit: a pulsed NMR approach. PMID- 17425049 TI - Metabolic activity of the central and peripheral regions of mesencephalic auditory center of reptiles. PMID- 17425050 TI - Muscular tissue of bony fish inhabiting water bodies located in the 30-kilometer zone around the Chernobyl nuclear power plant. PMID- 17425051 TI - Independent origin of the trichoptile and neoptile in bird plumage. PMID- 17425052 TI - Monogamy as one of the ways of realization of the adaptive potential in mammals as exemplified by the steppe lemming Lagurus lagurus Pallas. PMID- 17425053 TI - Protective effect of exogenous 70-kDa heat shock protein during endotoxic shock (sepsis). PMID- 17425054 TI - The first representative vertebrate fauna from the late miocene of southern European Russia. PMID- 17425055 TI - Stabilizing function of sexual selection in relation to male aggressiveness in rodents. PMID- 17425056 TI - Effect of exogenous ecdysteroids on growth, development, and fertility of the Egyptian cotton leafworm Spodoptera littoralis Boisd. (Lepidoptera: Noctuidae). PMID- 17425057 TI - Thermotolerance of mammalian cells is not induced near the restriction point of the G1 phase of the cell cycle. PMID- 17425058 TI - Iron chelator deferoxamine induces epithelial transformation of tumor cells. PMID- 17425059 TI - Obtaining hybrid mammalian cells containing diploid chromosome number. PMID- 17425060 TI - Beta-catenin does not show nuclear location in undifferentiated murine embryonic stem cells. PMID- 17425061 TI - How effective are lifestyle changes in the prevention of type 2 diabetes mellitus? AB - Obesity and impaired glucose tolerance are associated with a greater risk for a number of conditions, including insulin resistance, diabetes mellitus, hypertension, dyslipidemia, coagulation abnormalities, inflammatory markers, and coronary heart disease. Lifestyle changes can delay or prevent the development of type 2 diabetes in patients with obesity and impaired glucose tolerance. The risks improve with weight loss and increased physical activity. A decrease of 7% to 10% or more from baseline weight can have a significant effect. This has now been documented in a number of randomized, controlled studies. PMID- 17425062 TI - Cariogenic bacteria as biomarkers for sugar intake. AB - Cariogenic bacteria (lactobacilli and mutans streptococci) in saliva are possible biomarkers for sugar intake. We evaluated the strength of the evidence for an association between sugar intake and the two groups of bacteria, and concluded that there is a limited to moderately scientific basis for this association. Because of many confounding factors, bacteria counts can never be used as a precise measurement of sugar intake, but might still be useful on a group level. PMID- 17425063 TI - Leucine and protein synthesis: mTOR and beyond. AB - The effects of amino acid intake on protein synthesis in the intact rat appear to be mediated almost entirely by a single amino acid: leucine. The effect of leucine on protein synthesis appears to be closely associated with eIF4G phosphorylation and its association with eIF4E, but whether eIF4G phosphorylation actually mediates the effects of leucine or is merely associated with these events has not been elucidated. Additional research is needed to determine whether leucine effects eIF4G phosphorylation, whether eIF4G phosphorylation is essential for the effect of leucine on protein synthesis, and whether mTOR (mammalian target of rapamycin) or another component of the mTOR complex is somehow involved in leucine-specific signaling. PMID- 17425064 TI - Identification of a functional polymorphism at the adipose fatty acid binding protein gene (FABP4) and demonstration of its association with cardiovascular disease: a path to follow. AB - Fatty acid binding proteins (FABPs) are proteins that reversibly bind fatty acids and other lipids. So far, nine tissue-specific cytoplasmic FABPs have been identified. Adipose tissue FABP (FABP4) has been suggested to be a bridge between inflammation and other pathways related to the metabolic syndrome. In this regard, genetic variability at the FABP4 locus has been shown to be associated with plasma lipid levels, type 2 diabetes, and coronary heart disease risk. PMID- 17425065 TI - A novel solution is needed to correct low nutrient intakes in elderly long-term care residents. AB - Currently, there is no consensus about how to best treat malnutrition in elderly long-term care residents. A recent study found that the traditional treatment method of serving small, frequent meals was not effective at increasing energy intake. This review describes strategies to treat malnutrition in this vulnerable population. PMID- 17425066 TI - A recent brief critical review on how an increased intake of alpha-tocopherol can suppress the bioavailability of gamma-tocopher. PMID- 17425067 TI - Stormwater pollutant loads modelling: epistemological aspects and case studies on the influence of field data sets on calibration and verification. AB - In urban drainage, stormwater quality models have been used by researchers and practitioners for more than 15 years. Most of them were initially developed for research purposes, and have been later on implemented in commercial software packages devoted to operational needs. This paper presents some epistemological problems and difficulties with practical consequences in the application of stormwater quality models, such as simplified representation of reality, scaling up, over-parameterisation, transition from calibration to verification and prediction, etc. Two case studies (one to estimate pollutant loads at the outlet of a catchment, one to design a detention tank to reach a given pollutant interception efficiency), with simple and detailed stormwater quality models, illustrate some of the above problems. It is hard to find, if not impossible, an "optimum" or "best" unique set of parameters values. Model calibration and verification appear to dramatically depend on the data sets used for their calibration and verification. Compared to current practice, collecting more and reliable data is absolutely necessary. PMID- 17425068 TI - An integrated modeling approach to predict flooding on urban basin. AB - Correct prediction of flood extents in urban catchments has become a challenging issue. The traditional urban drainage models that consider only the sewerage network are able to simulate the drainage system correctly until there is no overflow from the network inlet or manhole. When such overflows exist due to insufficient drainage capacity of downstream pipes or channels, it becomes difficult to reproduce the actual flood extents using these traditional one-phase simulation techniques. On the other hand, the traditional 2D models that simulate the surface flooding resulting from rainfall and/or levee break do not consider the sewerage network. As a result, the correct flooding situation is rarely addressed from those available traditional 1D and 2D models. This paper presents an integrated model that simultaneously simulates the sewerage network, river network and 2D mesh network to get correct flood extents. The model has been successfully applied into the Tenpaku basin (Nagoya, Japan), which experienced severe flooding with a maximum flood depth more than 1.5 m on September 11, 2000 when heavy rainfall, 580 mm in 28 hrs (return period > 100 yr), occurred over the catchments. Close agreements between the simulated flood depths and observed data ensure that the present integrated modeling approach is able to reproduce the urban flooding situation accurately, which rarely can be obtained through the traditional 1D and 2D modeling approaches. PMID- 17425069 TI - Detection of gas pockets in pressurised wastewater mains using dynamic system response analysis. AB - In the Netherlands, wastewater is mostly collected in combined sewer system and transported to a WWTP through pressure mains. These pressure mains form an element of the system that did not receive much attention lately with respect to monitoring of performance and maintenance. For that reason, their state of functioning is often not known. Failure of operation is only noticed when during storm conditions the capacity of the system drops, resulting in undesirable discharge to the surface water. A recently made inventory showed that about half of the pressure mains show increased pressure loss for no obvious reason. Reduction of the systems nominal capacity can result from many causes, like increased wall roughness, scaling and the occurrence of free gas in the pipeline. The occurrence of gas-pockets may be caused by degassing of dissolved gas, but also by air entrapment at the pump inlet or at air valves. A research project is started that will be carried out from 2003 to 2005. This project focuses on three goals: the description of the gas-water transport phenomena in wastewater pressure mains, a method to detect and diagnose gas problems, and to overcome future problems by either applying remedial measures or by developing improved design methods for wastewater pressure systems. This paper describes the layout of the experimental set-up to detect and diagnose gas problems and the first results of the experiments are presented and discussed. The results are used to validate a model (Wanda) developed by Delft Hydraulics that describes the phenomena involved in surges in air/water transport. PMID- 17425070 TI - Implementing a space-time rainfall model for the Sydney region. AB - This paper investigates a Spatial Neyman-Scott Rectangular Pulse (SNSRP) model, which is one of only a few models capable of continuous simulation of rainfall in both space and time. The SNSRP is a spatial extension of the Neyman-Scott Rectangular Pulse model at a single point. The model is highly idealized having six parameters: storm arrival, cell arrival, cell radius, cell lifetime and two cell intensity parameters. A spatial interpolation of the scale parameter is used so that the model can be simulated continuously in space, rather than as a multi site model. The parameters are calibrated using least-squares fits to statistical moments based on data aggregated to hourly and daily totals. The SNSRP model is calibrated to a very large network of 85 gauges over metropolitan Sydney and shows a good agreement to calibrated statistics. A simulation of 50 replicates over the region compares favourably to several observed temporal statistics, with an example given for one site. A qualitative discussion of the simulated spatial images demonstrates the underlying structure of non-advecting cylindrical cells. PMID- 17425071 TI - Prediction of stormwater particle loads from impervious urban surfaces based on a rainfall detachment index. AB - This paper makes use of Non-Coarse Particle (NCP) data collected from three different impervious surfaces in Toowoomba, Australia. NCP is defined as suspended solids less than 500 microm in size. NCP loads (in mg/m(2)) were derived for 24 storms from a galvanized iron roof, a concrete car park and a bitumen road pavement. A scatter plot analysis was used to identify potential correlations between NCP loads and basic rainfall parameters such as rainfall depth and intensity. An exponential-type trend, consistent with many washoff models, was evident between load and average rainfall intensity for all surfaces. However, load data for some storms did not fit this general trend. Various indices, comprising different combinations of basic rainfall parameters, were evaluated as an alternative to rainfall intensity. A composite index, referred to as the Rainfall Detachment Index, was found to be better than average rainfall intensity in explaining a relationship between NCP load and storm rainfall characteristics. The selected rainfall index utilizes 6-minute rainfall intensities and is a variant of the well known Rainfall Erosivity Index (EI30) used for soil erosion estimation. PMID- 17425072 TI - The prediction of solute transport in surcharged manholes using CFD. AB - Solute transport processes occur within a wide range of water engineering structures, and urban drainage engineers increasingly rely on modelling tools to represent the transport of dissolved materials. The models take as input representative travel time and dispersion characteristics for key system components, and these generally have to be identified via field or laboratory measurements. Computational Fluid Dynamics (CFD) has the potential to reveal the underlying hydraulic processes that control solute transport, and to provide a generic means of identifying relevant parameter values. This paper reports on a study that has been undertaken to evaluate the feasibility of utilising a CFD based approach to modelling solute transport. Discrete phase modelling has been adopted, as this is computationally efficient and robust when compared with the time-dependent solution of the advection-dispersion equation. Simulation results are compared with published laboratory data characterising the dispersion effects of surcharged manholes, focusing specifically on an 800 mm diameter laboratory manhole for a flowrate of 0.002 m(3)/s and a range of surcharge depths. Preliminary indications are that the CFD results adequately replicate the measured downstream temporal concentration profiles, and that a threshold surcharge depth, corresponding to a change in hydraulic regime within the manhole, can also be identified. PMID- 17425073 TI - Application of transient storage modelling to solute transport across a surcharged manhole. AB - Results from previous solute tracer laboratory experiments across circular surcharged manhole structures by Guymer et al. have been used to optimise parameters within Hart's transient storage model (TSM). A surcharge threshold level for the model parameters is evident and this is explained in relation to jet theory. The ability to decompose the TSM is demonstrated with reference to frequency of exchange with the storage zone allowing the proportions of solute entering these regions to be inferred, together with an indication of storage volume retention times. PMID- 17425074 TI - Identification of sewer pipes to be cleaned for reduction of CSO pollutant load. AB - To reduce the CSO (Combined Sewer Overflow) pollutant discharge, one of the effective options is cleaning of sewer pipes before rainfall events. To maximize the efficiency, identification of pipes to be cleaned is necessary. In this study, we discussed the location of pipe deposit in dry weather in a combined sewer system using a distributed model and investigated the effect of pipe cleaning to reduce the pollutant load from the CSO. First we simulated the dry weather flow in a combined sewer system. The pipe deposit distribution in the network was estimated after 3 days of dry weather period. Several specific pipes with structural defect and upper end pipes tend to have an accumulation of deposit. Wet weather simulations were conducted with and without pipe cleaning in rainfall events with different patterns. The SS loads in CSO with and without the pipe cleaning were compared. The difference in the estimated loads was interpreted as the contribution of wash-off in the cleaned pipe. The effect of pipe cleaning on reduction of the CSO pollutant load was quantitatively evaluated (e.g. the cleaning of one specific pipe could reduce 22% of total CSO load). The CSO simulations containing pipe cleaning options revealed that identification of pipes with accumulated deposit using the distributed model is very useful and informative to evaluate the applicability of pipe cleaning option for CSO pollutant reduction. PMID- 17425075 TI - A performance investigation of small-bore sewers. AB - This paper describes a full-scale physical model and its application to investigate the effectiveness/performance of small-bore sewers for a range of operational and design parameters. The implementation methodology involves observing the movement of synthetic gross solids in three small bore sewers (150, 100 and 75 mm diameter) for different volumes of simulated flush waves and gradients. The simulated flush waves were generated, using an automated wave sequencer, for three different flush volumes (3, 4.5 and 6 litres). To investigate the impact of solid shape factor, a number of tests were carried out using synthetic solids in combination with toilet tissue paper. In total, more than 1,000 tests were performed for different operational and design parameter combinations. Results obtained to date have confirmed earlier studies, particularly with respect to the role of flush volume in solids transport, and identified the impact of gradient variation and its significance particularly in small-bore sewers receiving low flush volume. Results from the physical model application exercise will be used to propose new design guidelines for wastewater collection systems with specific consideration to new developments and inform the decision support system, currently being developed as part of a research project on water cycle management for new developments (WaND). PMID- 17425076 TI - Changes in isotope ratios during domestic wastewater production. AB - This paper presents considerations for the application of the natural water isotope method on catchment areas. For the estimation of the amount of infiltration and inflow in sewer systems the paper shows two applications in the Netherlands: one successful application on a relatively small catchment area with a simple geo-hydrological groundwater system and one unsuccessful application in an area that shows a large heterogeneity of delta 18O values in groundwater. Also, the paper focuses on the validity of the assumption that the isotopic ratios of drinking water are equal to those of strict domestic wastewater. In the transition from drinking water to strict wastewater it is shown that changes in isotopic composition of the water due to evaporation in common household appliances and effects inside the human body are insignificant. However, the presence of high-efficiency condensing boilers in an area can significantly influence the delta 18O value of strict wastewater, especially in winter months. This effect should be taken into account when applying the isotope method in such areas. PMID- 17425077 TI - The effect of rainfall measurement uncertainties on rainfall-runoff processes modelling. AB - Rainfall data are a crucial input for various tasks concerning the wet weather period. Nevertheless, their measurement is affected by random and systematic errors that cause an underestimation of the rainfall volume. Therefore, the general objective of the presented work was to assess the credibility of measured rainfall data and to evaluate the effect of measurement errors on urban drainage modelling tasks. Within the project, the methodology of the tipping bucket rain gauge (TBR) was defined and assessed in terms of uncertainty analysis. A set of 18 TBRs was calibrated and the results were compared to the previous calibration. This enables us to evaluate the ageing of TBRs. A propagation of calibration and other systematic errors through the rainfall-runoff model was performed on experimental catchment. It was found that the TBR calibration is important mainly for tasks connected with the assessment of peak values and high flow durations. The omission of calibration leads to up to 30% underestimation and the effect of other systematic errors can add a further 15%. The TBR calibration should be done every two years in order to catch up the ageing of TBR mechanics. Further, the authors recommend to adjust the dynamic test duration proportionally to generated rainfall intensity. PMID- 17425078 TI - Sewer solids separation by sedimentation--the problem of modeling, validation and transferability. AB - Sedimentation of sewer solids in tanks, ponds and similar devices is the most relevant process for the treatment of stormwater and combined sewer overflows in urban collecting systems. In the past a lot of research work was done to develop deterministic models for the description of this separation process. But these modern models are not commonly accepted in Germany until today. Water Authorities are sceptical with regard to model validation and transferability. Within this paper it is checked whether this scepticism is reasonable. A framework-proposal for the validation of mathematical models with zero or one dimensional spatial resolution for particle separation processes for stormwater and combined sewer overflow treatment is presented. This proposal was applied to publications of repute on sewer solids separation by sedimentation. The result was that none of the investigated models described in literature passed the validation entirely. There is an urgent need for future research in sewer solids sedimentation and remobilization! PMID- 17425079 TI - The effect of selection of time steps and average assumptions on the continuous simulation of rainwater harvesting strategies. AB - The use of domestic rainwater tanks with back up from mains water supplies in urban areas can produce considerable reductions in mains water demands and stormwater runoff. It is commonplace to analyse the performance of rainwater tanks using continuous simulation with daily time steps and average water use assumptions. This paper compares this simplistic analysis to more detailed analysis that employs 6 minute time steps and climate dependent water demand. The use of daily time steps produced considerable under-estimation of annual rainwater yields that were dependent on tank size, rain depth, seasonal distribution of rainfall, water demand and tank configuration. It is shown that analysis of the performance of rainwater tanks is critically dependent on detailed inputs. PMID- 17425080 TI - Achieving multiple benefits from stormwater harvesting. AB - As the concept of integrated urban water management is incorporated into the practice of urban water servicing, new options, such as stormwater harvesting, which can have multiple benefits, are of increasing interest. The multi functional benefits of stormwater harvesting include the potential to enhance urban stream health through improvements to the flow regime as well as providing a valuable water supply source. This paper synthesises a current research programme being undertaken to assess the viability of, and develop recommendations for, stormwater harvesting. The design of the collection, treatment, storage, flood protection, and distribution components of an integrated system are each discussed, along with the environmental flow consequences of urban stormwater harvesting. The incorporation of swales and biofilters into the collection system was not found to lead to significant exfiltration and evaporation losses in most circumstances and so can be employed as part of the treatment train. Further treatment can be provided by WSUD-type biophysical measures such as ponds, wetlands or novelly designed biofilters or physio-chemical treatment processes. Depending on the design, the stormwater storage component may or may not provide flood protection. In many circumstances, the storage capacity requirements are not considered to be a barrier to stormwater harvesting. PMID- 17425081 TI - Multicriteria procedure for the design and the management of infiltration systems. AB - Infiltration systems are frequently used as an option to manage urban storm drainage. By reducing flows and volumes in downstream sewers or in surface waters, they decrease the overflows and make it possible to recharge groundwater. They come in various forms with different uses; therefore, their performance is diverse and integrates multiple aspects. Consequently, a multicriteria approach was developed in order to quantify the performance of these systems and to help in decision making problems. For that purpose, a list of performance indicators integrating technical, economical, environmental and social aspects was developed. The performances were defined with the help of a working group composed of engineers from different technical or strategic departments from Greater Lyon and researchers from different fields. The paper presents the last version of the performance indicators tested according to a set of quality requirements: availability of data, relevance, fidelity, precision, sensitivity/robustness. This critical review of the set of indicators has led us to redefine a certain number of indicators, identify numerous biases and allow putting forward general instructions for criterion or indicator construction. The last phase is to propose multicriteria decision aid methods; a procedure using ELECTRE methods should be used. PMID- 17425082 TI - On the use of an ontology for the identification of degrees of freedom in urban wastewater systems. AB - The management of Urban Wastewater Systems (UWS) requires a comprehensive understanding of the interactions of processes and substances in the system. This leads to complex numerical models which can be applied to predict management actions or understand misconduction of the system. Nevertheless, for the communication between stakeholders in the process of optimizing a UWS these models are far too sophisticated. In this paper the use of an ontology is described serving as a conceptual model of the UWS which can be used for dissemination or as a preliminary stage to numerical models. The ontology therefore is part of a dissemination tool describing potential measures which can be applied to optimize a UWS. PMID- 17425083 TI - Development and verification of a general approach to describe the efficiency of vortex separators in combined sewer systems. AB - The water framework directive (CEC, 2000) asks for cost-effective measures for achieving good ecological conditions in receiving waters. Because of low operation costs and good pollutant removal efficiency, vortex separators (VS) are an attractive alternative to traditional stormwater tanks. The German design standard for CSO structures, ATV-A 128 (1992), demands long-term pollution load simulations. Today's simulation software, however, considers the removal processes in CSO structures either very rudimentarily or not at all. The higher pollutant removal efficiency of a structure like a VS cannot be taken into account. This might be one reason why VS are used still comparatively scarcely. A mathematical model describing the removal efficiency could increase the acceptance of VS. Several functions describing the removal efficiency have been derived from model tests or large-scale studies within the last few years. Within this paper, the data from three large-scale studies are used to verify the general applicability of one steady-state and one dynamic approach. The results show that the complex processes involved with CSO facilities and the large error related to monitoring make the validation of models a difficult task. Anyhow, especially the dynamic approach was applicable at all considered facilities. PMID- 17425084 TI - Nitrogen removal capacity of wetlands: sediment versus epiphytic biofilms. AB - Wetlands are important sinks for nutrients and constructed wetlands are current practice for stormwater treatment. For nitrogen, the main removal process is denitrification (microbial reduction of nitrate to nitrogen gas). The bacteria responsible for this process are mostly found in the sediments and in epiphytic biofilms growing on wetland macrophytes. This paper reports on a project which aimed at measuring denitrification potential in sediments and epiphyton in urban wetlands. This study showed that wetland sediments could support high rates of denitrification. Interestingly, the most polluted of the wetlands studied had the highest denitrification potential. The management implication from this result is that indicators of pollution, such as hydrocarbon levels, will not necessarily reflect the ability of a wetland to denitrify. Two of the wetlands were studied in more detail. Here the denitrification potential of the epiphyton on dominant macrophytes and sediments were measured. The results indicated that the potential denitrification activity of the epiphyton was comparable to those measured in the sediments. Hence, biofilms could play a significant part in removing nitrogen loads. This work contributes to a better knowledge of the functioning of wetlands. This will lead to improved design and management of wetlands used for treating stormwater. PMID- 17425085 TI - Removal of dissolved nitrogen, phosphorus and carbon from stormwater by biofiltration mesocosms. AB - Biofiltration systems are becoming a popular stormwater treatment device in water sensitive urban design for the removal of fine particulate and dissolved pollutants from stormwater. However, there is limited published data on the effectiveness of these systems for nutrient removal. We constructed biofiltration mesocosms to assess nutrient removal (nitrogen, phosphorus and carbon) under experimental conditions. Different types of media were compared (gravel, sand, and sandy-loam) in vegetated and non-vegetated mesocosms (six treatments in total). Five plant species were used. Vegetated sand and vegetated sandy-loam provided the best overall treatment. Vegetated mesocosms were very effective in removing nitrogen (63-77% removal) and phosphorus (85-94% removal) from synthetic stormwater, and removed substantially more nutrients than the non-vegetated treatments. All treatments removed a substantial portion of the carbon from the stormwater (28-66%). When flushed with tap water, nitrogen and phosphorus were retained by the vegetated mesocosms, but leached from the non-vegetated mesocosms. Plant growth was most vigorous in the sandy-loam media, indicating that this is a good growth media, even without the addition of organic matter. PMID- 17425086 TI - Local infiltration devices at parking sites--experimental assessment of temporal changes in hydraulic and contaminant removal capacity. AB - On site infiltration of stormwater is a common practice in order to avoid hydraulic overload of the urban drainage system. If hydrological conditions allow on-site infiltration--this is even mandatory from a legal point of view. Focus in this work is on surface infiltration of stormwater from parking lots. Proper operation of those devices is assumed to be appr. 15 years, as permits granted are limited to this time. Questions are raised whether this considered life expectancy is feasible. One apprehension is a possible clogging effect reducing the hydraulic capacity of the swale. The second aim was to identify magnitudes of accumulated pollutant loads with respect to limitations onto lifetime. The experimental investigation covered infiltration swales of different ages from eleven supermarket parking lots in Tyrol. Hydraulic permeabilities were assessed as well as chemical conditions of the soil material regarding hydrocarbon index (HI) and heavy metals (Cu, Zn, Pb and Cd). Further mass balance of contaminants has been performed in order to assess the operational life time based on pollutant load consideration. Calculations were based on load estimations using literature based minima and maxima concentrations from surface flows. Testing the correlation of hydraulic and pollutant measurements against site specific parameters (age, traffic load) revealed no distinct relation. In general all measured pollutants were found under limit concentrations. Mass balance calculations showed that limit concentrations are not exceeded either for worst case loading and considering 15 years of operation. PMID- 17425087 TI - Stormwater reuse: designing biofiltration systems for reliable treatment. AB - Stormwater reuse is increasing in popularity as a technique for overcoming water shortages in urban Australia. However, technology for the reliable treatment of stormwater for reuse is still not fully developed. This paper presents the first steps in refining biofilters for stormwater reuse. Six different filter media were selected, to target specific stormwater pollutants, as well as support plant growth. They were tested in the laboratory, where the filters were dosed three times per week with semi-synthetic stormwater for five weeks. Pollutant removal performance was monitored, and revealed that all soil-based filters performed similarly (while sand filters behaved somewhat differently). All filters removed more than 80% of solids and greater than 90% of lead, copper, and zinc. Three filter types were able to remove some phosphorus (particularly in the top 30 cm of the media). Apart from sand, all filter media were net producers of nitrogen, leading to an important conclusion that non-vegetated, soil-based filters are not suitable for targeting nutrients. However, since heavy metals are the primary pollutant of concern with respect to stormwater reuse for irrigation (the most popular end-use), it was concluded that biofilters may be promising technologies for treatment of stormwater for reuse. PMID- 17425088 TI - Development of a confined water sensitive urban design (WSUD) system using engineered soils. AB - Innovative Water Sensitive Urban Design (WSUD) systems are being investigated at three locations to the north and south of Sydney, Australia. These systems contain porous concrete pipes that are designed so that stormwater exfiltrates through the permeable walls of the pipes into the surrounding substrate media material. The porous pipes and media material treat the passing stormwater. The primary aim of the overall project is to develop a model to describe the treatment effectiveness of confined WSUD systems. This paper focuses on the system located at the Weathertex Industrial Site, Heatherbrae. Due to wood processing operations that occur at this site, it is recognised that the surface runoff will carry a heavy organics loading. Granulated Activated Carbon (GAC) is recognised for its ability to reduce the concentration of dissolved organics present in both wastewater and stormwater. GAC was therefore chosen as a filtration medium to be investigated at this site. To maximise the effectiveness of the GAC, extensive laboratory batch studies were undertaken prior to the field system being constructed to determine the optimum GAC/sand ratio. The purpose of the experimental work was to assess the dissolved organic removal potential through sorption of various concentrations of GAC. The aim of this paper is to describe these laboratory experiments and discuss how they related to the field system. Through these experiments it was determined that a sand/GAC ratio of 25:1 was ideal for the media material at the Heatherbrae site. PMID- 17425089 TI - Influence of compost characteristics on heavy metal sorption from synthetic stormwater. AB - This paper has the aim to assess the ability of garden derived compost to remove dissolved heavy metal contaminants typically found in stormwater. Compost was found to have excellent chemical and physical properties for the sorption of dissolved metal ions (Cu2+, Pb2+ and Zn2+). Batch sorption data were used to determine the sorption efficiency of Cu (93%), Zn (88%) and Pb (97%) by compost. The relative sorption affinity of these metals by compost is found to be in the order of Pb2+ > Cu2+ Zn2+. The effect of different particle size fractions of compost upon the sorption of Cu was also investigated. Sorption conformed to the linear form of the Freundlich isotherm and can be considered favourable because the sorption intensity values obtained in this study are between 0.1 and 1. Compost with a smaller particle size fraction has larger surface areas and greater sorption than the larger particle size fraction. Compost derived from garden waste is efficient for removal of heavy metals from wastewater or treating water for industries. PMID- 17425090 TI - A process-based method of estimating exponential decay parameters for particulate removal in a gravel trench. AB - Observed reductions in pollutant concentrations through stormwater treatment devices commonly display the characteristic form of exponential decay, in which the rate of decrease of pollutant concentration with distance is proportional to the concentration. The observation of an apparently irreducible or background pollutant concentration, C*, in many devices has led to development of the two parameter "k-C*" model. It is known that this model is too simplistic because the parameters k and C* are not constant but can vary greatly with pollutant concentration and hydraulic conditions. This paper presents an alternative exponential decay model for filtration of particulate pollutants, which is based on simple mathematical descriptions of key removal processes. The model delivers a process-based method for estimating the exponential decay constant. Moreover, the need to specify a background concentration is eliminated. To test the theory, the model is applied to the removal of clay and silica particles from horizontal flow through an experimental gravel trench. Particle concentrations were measured at nine locations along a 7.2 m long flume. The model agrees very well with the observed change in suspended solids concentration for the two pollutant materials and the range of flow rates tested. A single model parameter, notionally representing the "stickiness" of pollutant particles, is required for different pollutant materials. PMID- 17425091 TI - Long-term hydraulic and pollution retention performance of infiltration systems. AB - Infiltration techniques are now widely used to manage stormwater in urban areas. These techniques are used and recognized around the world for their many advantages, such as decreasing stormwater flow in sewer systems and recharging groundwater. But numerous cases of infiltration devices that failed after a few years of operation are still being reported. This study, which is based on site monitoring of operational infiltration systems, is part of the Field Observatory for Urban Water Management (OTHU). The main goals of this study are to improve knowledge of long-term hydraulic behaviour, especially as concerns the clogging speed and the quality of the runoff. This article will present the site, the monitoring process and the model that will be used to assess the hydraulic behaviour. First results of the calibration of the model show that the model is able to assess the hydraulic behaviour of the basin when it is clogged (average value of hydraulic resistance 17.1 h) and when it has been scraped (hydraulic resistance less than 3.8 h). However, further data are needed in order to validate the model. We also show that the experimental setup is well designed to assess the water volume and the sediment brought to the basin with low uncertainties. PMID- 17425092 TI - Identifying the major influences on the microbial composition of roof harvested rainwater and the implications for water quality. AB - Perceptions of the quality of roof harvested rainwater remain an impediment to widespread implementation of rainwater tanks on urban allotments. Previous literature reports on roof water quality have given little consideration to the relative significance of airborne environmental micro-organisms to roof catchment contamination and the issue of tank water quality. This paper outlines the findings of a recent study into the influence of weather on roof water contamination conducted at an urban housing development in Newcastle, on the east coast of Australia. Samples of direct roof run-off were collected during a number of separate rainfall events, and microbial counts were matched to climatic data corresponding to each of the monitored events. Roof run-off contamination was found to be under the strong influence of both wind speed and direction. The preliminary findings of an investigation currently under way into the microbial diversity of rainwater harvesting systems have also been presented. The results indicate that the composition of organisms present varied considerably from source to source and throughout the collection system. In all cases, evidence of faecal contamination was found to be negligible. The implications of these findings to the issues of tank water quality, health risk analysis and monitoring protocols have been discussed. PMID- 17425093 TI - Combined sewer system versus separate system--a comparison of ecological and economical performance indicators. AB - This paper aims at comparing the cost-effectiveness of the two main types of urban drainage systems, that is, the combined sewer system and the separate sewer system, based on the analysis of simulations. The problem of which of the two systems is better was heavily discussed over the years and the answer given to the question was usually: 'it depends'. In this work, specific impacts are investigated in terms of a cause-effect analysis. The results are subsequently summarized and can help in the choice of the system to be implemented. Despite earlier reasoning, studies on river water quality strongly indicate that the separate system is not always the preferable solution because the polluted runoff from the street, containing e.g. different heavy metals, is discharged directly into the river. This analysis aims to compare the two different sewer systems on the basis of literature data and simulation of specific cases. The results are evaluated, as suggested in the EU-Water Framework Directive, on the basis of different assessment criteria: river water quality and morphology impacts, emissions and costs. PMID- 17425094 TI - Is stormwater harvesting beneficial to urban waterway environmental flows? AB - Urbanization degrades the hydrology and water quality of waterways. Changes to flow regimes include increased frequency of surface runoff, increased peak flows and an increase in total runoff. At the same time, water use in many cities is approaching, and in some cases exceeding, sustainable limits. Stormwater harvesting has the potential to mitigate a number of these detrimental impacts. However, excessive harvesting of stormwater could also be detrimental to stream health. Therefore, a study was undertaken to test whether typical stormwater harvesting scenarios could meet the dual objectives of (i) supplying urban water requirements, and (ii) restoring the flow regime as close as possible to 'natural' (pre-developed). Melbourne and Brisbane, which have different climates, were used along with three land use scenarios (low, medium and high density). Modelling was undertaken for a range of flow and water quality indicators. The results show that using these typical harvesting scenarios helped to bring flow and water quality back towards their pre-developed levels. In some cases, however, harvesting resulted in an over-extraction of flow, demonstrating the need for optimizing the harvesting strategy to meet both supply and environmental flow objectives. The results show that urban stormwater harvesting is a potential strategy for achieving both water conservation and environmental flows. PMID- 17425095 TI - Sewer system design moving into the 21st century--a UK perspective. AB - Change in external factors, such as environmental legislation and climate change, will mean the future of sewerage systems is likely to be different from the past. Combined sewerage systems comprise the vast majority of existing sewers in countries such as the UK. A study funded by UK Water Industry Research Ltd has reviewed the current state of sewerage within the UK, the likely drivers for change and the consequent future impacts over a 75 year timescale. Potential responses to address the anticipated changes have also been considered. It is concluded that due to the wide extent and value of existing sewer systems, these will continue to be used for the foreseeable future. However, in order to meet the major challenges as a result of changing external factors, these need to be operated more effectively, new ideas need to be explored and moves to develop better and more integrated water management systems need to be started if sewer systems in the UK are to provide the anticipated required levels of service well into the 21st century. PMID- 17425096 TI - Understanding community receptivity to water re-use: Ku-ring-gai Council case study. AB - This social research project investigated community receptivity to using rainwater and greywater as alternative domestic water sources. It was focused in the Ku-ring-gai local government area in northern Sydney, and involved a household questionnaire followed by community leader interviews and resident focus groups. Trends, such as a prolonged drought and increasing population, compound the current crisis and concern facing Sydney's available water supply. Substitution of domestic potable water has been promoted as part of the solution. The research results revealed that community receptivity was highest for external uses, such as watering gardens and flushing toilets, and progressively decreased with increasing personal contact. Receptivity to greywater reuse fell more rapidly with the community believing there was a higher health risk associated with its use. Gender and cultural background were revealed as significant variables and give insight into the design of strategies to target these demographic groups. This evidence provides a reliable stocktake of current receptivity revealing that the community has good awareness and positive association with water reuse for many household activities. This now needs to be harnessed through programs targeted at developing skills, resources and motivation for new water reuse practices and technologies across diverse social groupings. PMID- 17425097 TI - Simultaneous identification and determination of major taxoids from extracts of Taxus chinensis cell cultures. AB - Liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI MS/ MS) and high-performance liquid chromatography (HPLC) methods have been used to identify and quantify the major taxoids from extracts of Taxus chinensis cell cultures. Chromatography was carried out on a reverse phase C18 column with isocratic-mode elution. By analytically comparing LC/ESI-MS/MS of the extracts with that of the available reference substances and literature, six taxoids were identified as taxuyunnanine C (Tc, 1), yunnanxane (2), 2alpha,5alpha10beta triacetoxy-14beta-propionyloxytaxa-4(20),11-diene (3), 2alpha,5alpha, 10beta triacetoxy-14beta-(2-methyl)butyryloxytaxa-4(20),11-diene (4), taxol (5), and baccatin III (B III, 6), respectively. Among them, 2, 3 and 4 were assigned in the absence of the corresponding reference substances, and 3 and 4 were detected in this cell line for the first time. The identification was validated by NMR spectra. The precise quantification of 1 and 5 was made using HPLC. The limit of detection (LOD), 0.5 microg/ml for 5, 1.5 microg/ml for 1, and the linearity and accuracy of the quantitative method were evaluated, indicating a wide linear range and satisfactory accuracy. The amounts of other identified taxoids were calculated on the basis of comparison of the absolute response factors of similar structural substances. The proposed method provides a rapid, conventional and reliable tool to characterize and study cell lines for elucidating the taxane biosynthesis. PMID- 17425098 TI - Phomopsidone, a novel depsidone from an endophyte of the medicinal plant Eupatorium arnottianum. AB - The medicinal plant Eupatorium arnottianum can be found in the Northeast and center of Argentina and the South of Bolivia. From plant material collected in Argentina an endophytic Phomopsis was isolated. The fungus was identified by microscopic features and analysis of its ITS sequence. Cultures yielded, besides mellein and nectriapyrone, a novel depsidone derivative for which we propose the name phomopsidone (1). The structure of 1 was determined from its spectroscopic data. PMID- 17425099 TI - Narcotic alkaloids of four papaver species from Iran. AB - Four native Papaver species of Iran, i. e. P. glaucum, P. tenuifolium, P. dubium and P. fugax, were collected from their natural habitat and subjected to HPLC analysis for determination of their morphine, codeine and thebaine content. P. dubium and R. glaucum contained all of the three mentioned narcotic alkaloids, while morphine was not found in P. fugax, and P. tenuifolium was free from codeine. PMID- 17425100 TI - Antiviral and antimicrobial profiles of selected isoquinoline alkaloids from Fumaria and Corydalis species. AB - In the current study, 33 isoquinoline alkaloids belonging to protopine-, benzylisoquinoline-, benzophenanthridine-, spirobenzylisoquinoline-, phthalideisoquinoline-, aporphine-, protoberberine-, cularine-, and isoquinolone types as well as 7 derivatives of them obtained from some Fumaria and Corydalis species growing in Turkey have been evaluated for their in vitro antiviral and antimicrobial activities. Both DNA virus Herpes simplex (HSV) and RNA virus Parainfluenza (PI-3) were employed for antiviral assessment of the compounds using Madine-Darby bovine kidney and Vero cell lines and their maximum non-toxic concentrations (MNTC) and cytopathogenic effects (CPE) were determined using acyclovir and oseltamivir as the references. Antibacterial and antifungal activities of the alkaloids were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis, and Candida albicans by the microdilution method and compared to ampicilline, ofloxacine, and ketocanazole as the references. The alkaloids did not present any notable antibacterial effect, while they had significant antifungal activity at 8 microg/ml concentration. On the other hand, the alkaloids were found to have selective inhibition against the PI-3 virus ranging between 0.5 and 64 microg/ml as minimum and maximum CPE inhibitory concentrations, whereas they were completely inactive towards HSV. PMID- 17425101 TI - New anti-inflammatory agents. AB - The pyrrole derivatives la, b and 2a, b were used as precursors for the preparation of N-substituted pyrrole derivatives 3a, b-9a, b and pyrrolo[2,3 d]pyrimidines 13-16. Also, all the newly prepared products were tested for anti inflammatory activity as analogues to fenamates, and some of them revealed moderate anti-inflammatory activity compared to the standard drug indomethacin. PMID- 17425102 TI - Polyphenols of Rosa L. leaves extracts and their radical scavenging activity. AB - Antioxidant potential of Rosa L. leaves methanolic extracts was evaluated in vitro using a spectrophotometric method based on measuring the radical scavenging effect on 2,2-di-phenyl-1-picrylhydrazyl (DPPH) radicals. The contents of ellagic acid, quercetin and kaempferol in the extracts from leaves of seventeen rose species were determined using SPE-RP-HPLC methods. Additionally, total phenolic content was determined spectrophotometrically according to the Folin-Ciocalteu procedure and calculated as gallic acid equivalents (GAE). Remarkable high antioxidant activity and high total phenolic content (5.7% < GAE < 15.2%), large ellagic acid (EA) content from 9.37 to 19.42 mg/g of dry weight, a quercetin content ranging from 3.68 to 15.81 mg/g of dry weight and kaempferol content from 1.25 to 9.41mg/g of dry weight were found in rose leaves. Significant correlation between EA (r(2) = 0.6131), quercetin (r(2) = 0.5158), total phenolic content (r(2)= 0.8485) and antioxidant activity was observed. Basing on the studies conducted one may assume that the extracts of rose leaves are a rich source of natural antioxidants and could be used to prevent free-radical-induced deleterious effects. PMID- 17425103 TI - Antinociceptive activity of structural analogues of rotundifolone: structure activity relationship. AB - Rotundifolone, a monoterpene isolated from the essential oil of the leaves of Mentha x villosa, is a constituent of several essential oils and known to have antinociceptive activity. Our recent study demonstrated that the analogues of rotundifolone showed also a significant antinociceptive effect. In the present report, to investigate the correlation between the structure and antinociceptive activity, rotundifolone and its analogues were evaluated in the acetic acid induced writhing test in mice. All compounds showed to be more antinociceptive than rotundifolone against the pain response induced by acetic acid. Comparing the antinociceptive effect of rotundifolone with limonene oxide and (+)-pulegone, the results demonstrated that the epoxide group contributes as much as the ketone group to the antinociceptive activity of rotundifolone. Similarly, pulegone oxide and carvone epoxide were more antinociceptive than rotundifolone, thereby suggesting that the position of the functional group on the ring also influences the antinociceptive activity. (-)-Carvone produced maximal inhibition of the writhing response and was slightly more active than (+)-carvone. The study showed that by appropriate structural modification it may be possible to develop novel antinociceptive agents. PMID- 17425104 TI - Root cultures of linum species section Syllinum as rich sources of 6 methoxypodophyllotoxin. AB - Linum spp. from section Syllinum are promising for the production of aryltetralin lignans like podophyllotoxin (PTOX) and 6-methoxypodophyllotoxin (MPTOX). MPTOX is a PTOX congener that has cytotoxic activity comparable with PTOX. In this study root cultures of Linum Bungei from section Dasyllinum, L. strictum from section Linastrum, L. album, L. mucronatum ssp. mucronatum and L. nodiflorum from section Syllinum were established and their MPTOX levels were investigated in 1000 ml flasks. Root cultures of L. mucronatum ssp. mucronatum and L. nodiflorum were used to examine cell growth and production of MPTOX during a culture period of 36 days in 250 ml flasks. Considerable amounts of MPTOX in root cultures (1000 ml flasks) of L. album (6 mg/100 g DW), L. mucronatum ssp. mucronatum (770 mg/100 g DW) and L. nodiflorum (91 mg/100 g DW) were detected while it wasn't detected in root cultures of L. Bungei and L. strictum. In time course experiments, the maximum amount of MPTOX in L. nodiflorum root culture was at day 16 with 480 mg/ 100 g DW and the maximum amount of MPTOX in L. mucronatum ssp. mucronatum root culture was at day 12 with 130 mg/100 g DW. The results showed that root cultures of Linum species from section Syllinum are rich sources of MPTOX and since this lignan has remarkable cytotoxic activity, it can be used as a precursor for the production of antitumor agents. PMID- 17425105 TI - Induction of phenolic compounds in wheat (Triticum aestivum L.) tissue cultures by streptomycin. AB - The tissue cultures of wheat (Triticum aestivum L.) were induced from the mature embryos (explants) of the dry grains and grown on MS medium containing kinetin (0.1 mg/1) and 2,4 D (1.0 mg/l). The cultures were incubated for two weeks at (25+/-2) degrees C under a light/dark regime (16 h light daily). The formed calli were subcultured at the beginning of the stationary growth phase (15 days) with fresh MS medium containing 0, 5, 10, 25, 50, 100, 150 mg/l streptomycin elicitor and maintained for two weeks for three subcultures. A significant increase in phenylalanine ammonia lyase (PAL) activity coincided with the increase of the total phenolic compounds after elicitation with streptomycin. Maximum induction was recorded during the first two weeks, then gradually declined during the rest of the experimental period, but the values attained were still markedly higher than that of the control. The endogenous cinnamic acid content was also increased significantly with the increase in PAL activity making about 2-18% of the total phenolic acids. The growth and accumulation of phenolic compounds were inversely related. However, accumulation of phenolic compounds became limited for growth of wheat tissue culture especially during the long term cultivation. PMID- 17425106 TI - The influence of plant hormones on phospholipid monolayer stability. AB - The influence of hormones in water subphase on the stability of monolayers built of phospholipid mixtures extracted from embryogenic (PLE) and nonembryogenic (PLNE) wheat calli was examined. Additionally, experiments on individual lipids, dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidic acid (DPPA), were performed. DPPC was chosen because it was the main phospholipid present in both calli. Negatively charged DPPA could mimic a negatively charged natural mixture of lipids. As hormones, auxins (IAA and 2,4-D), cytokinins (zeatin and kinetin) and zearalenone were chosen. The time of monolayer stability for PLNE calli was much longer than for PLE calli. Kinetics of monolayer stability of PLNE was similar to DPPA, whereas that of PLE was similar to DPPC. Generally, hormones increased the time after which the monolayer stability was reached and decreased the surface pressure. The greatest effect was observed for auxins (especially IAA), whereas cytokinins affected the monolayer stability to a lesser degree. PMID- 17425107 TI - Scoparone inhibits ultraviolet radiation-induced lipid peroxidation. AB - Antioxidant capabilities of scoparone, the component of Artemisia scoparia and other medicinal plants, against lipid peroxidation induced by ultraviolet radiation or Fenton reaction have been analyzed. Lipid peroxidation was monitored by measuring the absorption spectra of the conjugated dienes and quantified by the Klein oxidation index. Obtained results imply that scoparone is a very efficient inhibitor of ultraviolet radiation-induced lipid peroxidation and damage. PMID- 17425108 TI - Ultracytochemical localization of acid phosphatase in Humicola lutea conidia and mycelia. AB - Electron microscopic cytochemical procedures were used to determine the cellular location of acid phosphatase in the fungus Humicola lutea grown in casein containing medium lacking in mineral orthophosphates. In our investigations acid phosphatase in nongerminating conidia was localized on the outer side of the cell wall, in the cell wall, and on the exterior surface of the plasma membrane. The reaction product of acid phosphatase in germinating conidia was seen in the outer wall layer while in young mycelium on the cell surface and in the exocellular space. The relationship between phosphatase activities localized in the cell wall and their role in the enzymatic degradation of the phosphoprotein casein providing available phosphates for cell growth is discussed. PMID- 17425109 TI - Effect of copper on acid phosphatase activity in yeast Yarrowia lipolytica. AB - Acid phosphatase (APase) activity of the yeast Yarrowia lipolytica increased with increasing Cu2+ concentrations in the medium. Furthermore, the enzyme in soluble form was stimulated in vitro by Cu2+, Co2+, Ni2+, Mn2+ and Mg2+ and inhibited by Ag+ and Cd2+. The most effective ion was Cu2+, especially for the enzyme from cultures in medium containing Cu2+, whereas APase activity in wall-bound fragments was only slightly activated by Cu2+. The content of cellular phosphate involving polyphosphate was decreased by adding Cu2+, regardless of whether or not the medium was rich in inorganic phosphate. Overproduction of the enzyme stimulated by Cu2+ might depend on derepression of the gene encoding the APase isozyme. PMID- 17425110 TI - Copper extrusion after accumulation during growth of copper-tolerant yeast Yarrowia lipolytica. AB - The Cu2+-tolerant yeast Yarrowia lipolytica accumulated Cu2+ until the late logarithmic phase. Thereafter, Cu2+ was temperature-dependently extruded into phosphate-limited culture medium containing high concentrations of heavy metal ions but not into 10 mM 2-(N-morpholino)ethane sulfonic acid (MES) buffer (pH 6.0). Peptone in the culture medium played an important role in the extrusion, which proceeded even when peptone was substituted with cysteine or histidine, but not with any other amino acid tested. PMID- 17425111 TI - Influence of various phenolic compounds on phenol hydroxylase activity of a Trichosporon cutaneum strain. AB - The phenol-degrading strain Trichosporon cutaneum R57 utilizes various aromatic and aliphatic compounds as a sole carbon and energy source. The intracellular activities of phenol hydroxylase [EC 1.14.13.7] of a Trichosporon cutaneum R57 strain grown on phenol (0.5 g/l) were measured. Different toxic phenol derivatives (cresols, nitrophenols and hydroxyphenols) were used as substrates in the reaction mixture for determination of the enzyme activity. The data obtained showed that the investigated enzyme was capable to hydroxylate all applied aromatic substrates. The measured activities of phenol hydroxylase varied significantly depending on the aromatic compounds used as substrates. The rate of phenol hydroxylase activity with phenol as a substrate (1.0 U/mg total cell protein) was accepted as 100%. PMID- 17425112 TI - Bacterial decolorization of acid orange 7 in the presence of ionic and non-ionic surfactants. AB - The effects of the non-ionic surfactant Triton X-100, the cationic surfactant cetyltri-methylammonium bromide (CTAB) and the anionic surfactant sodium N lauroyl sarcosinate (SLS) on the decolorization of the reaction medium containing the monoazo dye Acid Orange 7 (AO7) by Alcaligenes faecalis and Rhodococcus erythropolis were studied. It was found that the surfactants influenced in different ways the rate of decolorization. At all concentrations tested the non ionic surfactant Triton X-100 decreased the decolorization rate of R. erythropolis. At concentrations above the critical micelle concentration (CMC) Triton X-100 upset the usually observed exponential decay of the dye with A. faecalis due probably to the existence of an outer membrane in this organism. In concentrations above the CMC the anionic surfactant SLS inhibited the decolorization and, at prolonged incubation, caused partial release of the bound dye. The cationic surfactant CTAB in concentrations above and below the CMC accelerated drastically the binding of AO7 to the cells causing a rapid staining of the biomass and complete decolorization of the reaction medium. An attempt was made for explanation of the observed differences by the negative electrostatic charge of the living bacterial cell. PMID- 17425113 TI - Inhibition of anodic galvanotaxis of green paramecia by T-type calcium channel inhibitors. AB - Calcium ion (Ca2+) is one of the key regulatory elements for ciliary movements in the Paramecium species. It has long been known that members of Paramecium species including green paramecia (Paramecium bursaria) exhibit galvanotaxis which is the directed movement of cells toward the anode by swimming induced in response to an applied voltage. However, our knowledge on the mode of Ca2+ action during green paramecia anodic galvanotactic response is still largely limited. In the present study, quantification of anodic galvanotaxis was carried out in the presence and absence of various inhibitors of calcium signaling and calcium channels. Interestingly, galvanotactic movement of the cells was completely inhibited by a variety of Ca2+-related inhibitors. Such inhibitors include a Ca2+ chelator (EGTA), general calcium channel blockers (such as lanthanides), inhibitors of intracellular Ca2+ release (such as ruthenium red and neomycin), and inhibitors of T-type calcium channels (such as NNC 55-0396, 1-octanol and Ni2+). However, L type calcium channel inhibitors such as nimodipine, nifedipine, verapamil, diltiazem and Cd2+ showed no inhibitory action. This may be the first implication for the involvement of T-type calcium channels in protozoan cellular movements. PMID- 17425114 TI - Isolation and identification of potential phosphate solubilizing bacteria from the rhizoplane of Oryza sativa L. cv. BR29 of Bangladesh. AB - A total of 30 bacteria were isolated from the rhizoplane of rice cv. BR29 cultivated in Mymensingh, Bangladesh and from the seedlings obtained from surface sterilized seeds of BR29. Upon screening, 6 isolates showed varying levels of phosphate solubilizing activity in both agar plate and broth assays using National Botanical Research Institute's phosphate medium. The bacterial isolates were identified based on their phenotypic and 16S rRNA genes sequencing data as Acinetobacter sp. BR-12, Klebsiella sp. BR-15, Acinetobacter sp. BR-25, Enterobacter sp. BR-26, Microbacterium sp. BRS-1 and Pseudomonas sp. BRS-2. The BR-25 exhibited highest phosphate solubilizing activity followed by BR-15. They grew rapidly in the liquid medium at pH 5 and 7 but almost no growth occurred at pH 3. The pH value of the culture medium was decreased with bacterial growth suggesting that they might secrete organic acids to solubilize insoluble phosphorus. Scanning electron microscope analysis of two-week-old rice seedlings germinated from seeds previously inoculated with BR-25 and BR-15 revealed dense colonization at the root surfaces presumably using fimbriae on the bacterial cells. PMID- 17425115 TI - Excess boron reduces polyphenol oxidase activities in embryo and endosperm of maize seed during germination. AB - The effects of increasing concentrations of boron (0, 0.1, 1, 10 and 20 mM) as boric acid on the rate of germination and polyphenol oxidase activities in embryo and endosperm tissues of maize seeds (Zea mays L. cv. Arifiye) were studied. The germination percentage of maize seeds was not affected by boron concentrations up to 10 mM, and decreased by 20 mM. Distilled water and lower boron concentrations (0.1 and 1 mM) increased polyphenol oxidase activities at the beginning of germination up to 12 h whereas its excess levels (10 and 20 mM) decreased polyphenol oxidase activities in embryos and endosperm during germination. Polyphenol oxidase activities with o-diphenolic substrates (caffeic acid, catechol and dopa) were found to be higher than with a monophenolic substrat (tyrosine) in both embryos and endosperms. Further, caffeic acid oxidizing polyphenol oxidase was found to show more activity in embryos of the seeds germinating in distilled water when compared to other substrates. PMID- 17425116 TI - Genetic variability and relationships among thirty genotypes of finger millet (Eleusine coracana L. Gaertn.) using RAPD markers. AB - Ragi or finger millet (Eleusine coracana L.) is an important crop used for food, forage, and industrial products. It is distributed in tropical and temperate regions of the world. The germplasm identification and characterization is an important link between the conservation and utilization of plant genetic resources. Traditionally, species or varieties identification has relied on morphological characters like growth habit, leaf architecture or floral morphology. Investigation through RAPD (random amplified polymorphic DNA) markers was undertaken for identification and determination of the genetic variation among thirty genotypes of ragi of the family Poaceae. Thirteen selected decamer primers were used for genetic analysis. A total of 124 distinct DNA fragments ranging from 300-3000 bp was amplified by using selected random RAPD marker. The genetic similarity was evaluated on the basis of the presence or absence of bands. Cluster analysis was made by the similarity coefficient. It indicated that the 30 genotypes of ragi form two major clusters, first, a major cluster having only one genotype, i. e. Dibyasinha and a second major cluster having twenty-nine genotypes. The second major cluster again subdivides into two minor clusters. A first minor cluster has only three varieties, i. e. Neelachal, OEB-56 and Chilika. The genotypes Neelachal and OEB-56 exhibit a 86% similarity with each other and 80% similarity with Chilika. A second minor cluster has 26 genotypes and is divided into two sub-minor clusters. The first sub-minor cluster has only one genotype (VL-322). The second sub-minor cluster again subdivides into two groups. One group has one genotype and the second group again is divided into two sub-groups, one with 13 genotypes and the other with 11 genotypes. The highest similarity coefficient was detected in a genotype collected from southern India and the least from northern India. The genotypes of finger millet collected from diverse agroclimatic regions of India constitute a wide genetic base. This is helpful in breeding programs and a major input into conservation biology of cereal crop. PMID- 17425117 TI - Nuclear-localized plastid DNA fragments in protozoa, metazoa and fungi. AB - We analyzed nuclear-localized plastid-like DNA (nupDNA) fragments in protozoa, metazoa and fungi. Most eukaryotes that do not have plastids contain 40-5000 bp nupDNAs in their nuclear genomes. These nupDNA fragments are mainly derived from repeated regions of plastids and distribute through the whole genomes. A majority of nupDNA fragments is located on coding regions with very important functions. Similar to plastids, these nupDNAs most possibly originate from cyanobacteria. Analysis of them suggests that through millions of years of universal endosymbiosis and gene transfer they may have occurred in ancient protists before divergence of plants and animals/fungi, and some transferred fragments have been reserved till now even in modern mammals. PMID- 17425118 TI - Binding of FKBP23 to BiP in ER shown by gel filtration chromatography. AB - FKBP23 was found in mouse endoplasmic reticulum (ER) in 1998. It consists of an N terminal peptidyl-prolyl cis/trans isomerase (PPIase) domain and a C-terminal domain with Ca2+ binding sites. Previously, we reported that FKBP23 specifically binds to BiP, the main protein of the molecular chaperone Hsp70 in ER lumen, and the binding is interrelated with the Ca2+ concentration. In this work we have found the existence of the complex FKBP23/BiP by separation of an ER extract using gel filtration chromatography (GFC), and that the existence of this complex is Ca2+-interrelated. This result further verified the Ca2+-interrelated binding of these two proteins in vivo. PMID- 17425119 TI - Identification of minor sex pheromone components of the poplar clearwing moth Paranthrene tabaniformis (Lepidoptera, Sesiidae). AB - A chemical analysis of the crude sex pheromone gland extracts of virgin calling Paranthrene tabaniformis females, obtained from the European part of Kazakhstan, revealed the presence of five compounds: (3E,13Z)-octadeca-3,13-dien-1-ol (E3,Z13 18:OH), (3Z,13Z)-octadeca-3,13-dien-1-ol (Z3,Z13-18:OH), (2E,13Z)-octadeca-2,13 dien-1-ol (E2,Z13-18:OH), (13Z)-octadec-13-en-1-ol (Z13-18:OH), and octadecan-1 ol (18:OH) at the ratios 64.0:32.4: 1.4:0.9:1.3, which are structurally related to sex pheromone components of clearwing moths. Our previous field tests showed synergistic effects of Z3,Z13-18:OH and E2,Z13-18:OH to attract P. tabaniformis males, when these compounds were tested in binary mixtures with the known sex pheromone E3,Z13-18:OH. The three dienic alcohols should all be considered as sex pheromone components of the P. tabaniformis species, while the role of Z13-18:OH and 18:OH remained unclear. PMID- 17425120 TI - Anisaldehyde, a melanogenesis potentiator. AB - Anisaldehyde (4-methoxybenzaldehyde), previously reported as a tyrosinase inhibitor, did not inhibit melanogenesis in cultured B16-F10 melanoma cells but rather enhanced it. This adverse effect of anisaldehyde was accompanied by melanocytotoxicity in a dose-dependent manner up to 2 mM. The melanin content per cell at 1 mM was increased 5-fold compared to control and morphological observations showed the deposition of melanin pigments. Anisaldehyde was also examined against cultured human A375 melanoma cells. PMID- 17425121 TI - New findings about Ellman's method to determine cholinesterase activity. AB - The original Ellman's spectrophotometrical method for cholinesterase activity determination uses 5,5'-dithiobis-2-nitrobenzoic acid (DTNB, Ellman's reagent) as a chromogen and records the level of cholinesterase activity as an increase of absorbance at 412 nm. Although this procedure usually poses no problem, exceptions arise when the concentration of DTNB is far higher than the concentration of acetylthiocholine (ATCH). It was found that the ratio of concentrations of DTNB/ATCH is an important parameter for the ATCH hydrolysis course: high excess of DTNB decreases the hydrolysis rate resulting in a lower measured enzyme activity. Our experiments indicate that this influence of DTNB concentration can be explained by the inhibition of ATCH hydrolysis by DTNB. PMID- 17425123 TI - Now and then: lost causes and new hope. PMID- 17425124 TI - Nurse led sedation for paediatric MRI: progress and issues. AB - AIM: This article describes the current magnetic resonance imaging (MRI) sedation service, the role of the nurse sedationist, results of clinical audits, and future issues within MRI. METHOD: Data from 2004 until 2006 were analysed to describe the percentage of children who were successfully sedated, and any complications. The recovery profile was investigated by a questionnaire of 100 consecutive cases. FINDINGS: Sedation was attempted in 455 cases with chloral hydrate, and 325 with temazepam and droperidol. The success rate was 97.4 and 92.6 per cent respectively. Top-up sedation was used in 10 and 29 per cent respectively. There were seven minor incidents but none required admission. Approximately 20 per cent of children were drowsy the following day. CONCLUSION: We believe that the success of the service depends upon three main factors: the deselection of children in whom sedation is unsafe or likely to be unsuccessful, the use of appropriate drugs in limited doses, and the training of experienced nurses. PMID- 17425125 TI - Intestinal transplantation in children: a new challenge for nursing. AB - Small bowel transplantation is a recent innovation and thus a relatively new area of nursing practice. There is a lack of literature on the nursing care of children whose only hope of long-term survival is small bowel transplantation. The aim of this article is to raise awareness about intestinal transplantation in children using a case study to highlight the main nursing implications including: multidisciplinary team working, monitoring fluid balance, caring for immunosuppressed patients, monitoring for acute rejection and discharge preparation of the child and family. PMID- 17425126 TI - Guided imagery and distraction therapy in paediatric hospice care. AB - Available evidence indicates deficits in the pain management of children with analgesic drugs used outside licensed boundaries, in situations where their pharmacokinetics are untested. A case series is used to demonstrate the effectiveness of more holistic techniques such as guided imagery and distraction therapy in reducing the pain experienced by children in the hospice setting. Using these techniques it may be possible to reduce the amount of analgesia or the frequency of administration. PMID- 17425127 TI - Nasojejunal tube placement in paediatric intensive care. AB - Nasojejunal delivery of enteral feeds is a safe and effective alternative to parenteral nutrition in critically ill children in whom intra-gastric feeding is usually poorly tolerated. A guideline for bedside placement of nasojejunal tubes (NJTs) was developed by a mulit-disciplinary group. An audit of practice was carried out following implementation of the guideline. During the audit period 27 NJTs were successfully passed in 21 patients. The result of this innovation has been early initiation of nasojejunal feeding and an increase in bedside placement of NJTs within the PICU. Paediatric radiologists have reported a reduction in requests for NJT placement under X-ray screening and there has been a reduction in the use of medication and X-ray to place NJTs. Based on the audit data, 58 per cent of the children would have definitely or probably commenced parenteral nutrition had NJT placement and feeding been unsuccessful. The audit also demonstrated that 26 out of 27 nurses and doctors reported they found the guidelines easy or very easy to follow. Reducing variations in practice through the use of guidelines increases the frequency of jejunal feeding. This benefits critically ill patients by improving tolerance of enteral feeding for better nutritional outcomes. PMID- 17425128 TI - Resilience in Afghan children and their families: a review. AB - Resilience enables individuals to survive the horrors of war and other such tragedies. This article draws on personal reflections of living in the post Taliban period in Afghanistan and a review of the literature to explore resilience processes that may be present in the Afghan population. Adverse factors that Afghan children and families face include: destruction of infrastructure, danger, traumatic experiences, post traumatic stress disorder, political and ideological commitment. The protective factors that may have enabled them to survive, and subsequently rebuild in the face of multiple challenges are absolute faith in Allah (God), family support and community support. Understanding more about the kinds of adversities they face and the nature of their protective processes gives healthcare professionals a basis for working with them to improve their health and wellbeing. PMID- 17425129 TI - From hospital to home for technology-dependent children. PMID- 17425130 TI - Writing for publication: a personal view. AB - Each year nurses attend study days and courses in order to keep up to date, improve their practice and meet NMC requirements for continuing professional development. When they return to work their new knowledge is often shared with colleagues to make a difference to patient care. This sharing of knowledge may change things at a local level but nurses need to consider disseminating local good practice on a wider scale. This can be achieved by writing for publication and presenting at conferences. PMID- 17425131 TI - Intramuscular injection technique. PMID- 17425132 TI - Contemporary aspects of infant feeding. AB - Early dietary influences, even before birth, can have an effect on growth, which in turn has a profound influence on adult health. Formula-fed infants have a different growth pattern from that of breast-fed babies and a greater risk of developing obesity. However, infant formulas, when correctly prepared and handled, are a safe alternative for mothers who do not wish to or are unable to breast feed. A balanced view of the nutritional benefits of breast milk and infant feed formulas allows professionals to share sound advice with mothers. PMID- 17425133 TI - A comparison of the performance of two chromatographic and three extraction techniques for the analysis of PAHS in sources of drinking water. AB - The aim of this work is to establish a sensitive and reliable method for the analysis of the 16 priority Environmental Protection Agency-defined polycyclic aromatic hydrocarbons (PAHs) found in water samples. Gas chromatography (GC)-mass spectrometry (MS) and high-performance liquid chromatography (HPLC)-fluorescence detection (FLD)-UV techniques are optimized to obtain an adequate resolution of all compounds. Validation of the methods is carried out, and a good performance is observed for both techniques. The HPLC-FLD-UV technique is somewhat more sensitive than the GC-MS technique for the determination of PAHs; thus, the HPLC FLD-UV method is used to follow up both the solid-phase extraction (SPE) analysis using cartridges and discs and the liquid-liquid extraction (LLE), which are also evaluated for the extraction of the PAHs. Low recoveries between 43% and 79% are obtained using SPE cartridges, and higher values are obtained using SPE discs (56 96%) and LLE (60-105%). Better results are obtained using the LLE technique, and, thus, analysis of real water samples is carried out using this technique. LODs between 0.6 and 21 ng/L and relative standard deviations less than 15% are obtained using a spiked water sample analyzed using the full LLE HPLC-FLD-UV method. PMID- 17425134 TI - Separation, characterization, and quantitation of process-related substances of the anti-hypertensive drug doxazosin mesylate by reversed-phase LC with PDA and ESI-MS as detectors. AB - A simple and rapid reversed-phase liquid chromatography (LC) method with photodiode array (PDA) and electrospray ionization (ESI)-mass spectrometry (MS) as detectors was developed and validated to separate, identify, and quantitate the related substances of Doxazosin mesylate (DXZN) for monitoring the reactions involved during process development. The high-performance liquid chromatography profiles of related-substances of DXZN are used as fingerprints to follow the procedures used in the manufacturing units. The separation is accomplished on an Inertsil ODS-3 column with acetonitrile-ammonium acetate (10 mM, pH 4.0) as the mobile phase, using a gradient elution mode and monitoring the eluents by a photodiode array detector at 265 nm at ambient temperature. LC-ESI-MS-MS is used to identify the additional impurities formed during the synthesis. The identified impurities were synthesized and characterized by UV, Fourier transform-IR, 1H NMR, and MS data. The detection limits for the impurities are 0.74 - 4.14 x 10( 9) g, and the method is found to be suitable not only for the monitoring of synthetic reactions, but also for quality assurance of DXZN in bulk drugs and formulations. PMID- 17425135 TI - Liquid chromatographic analysis of glucosamine in commercial dietary supplements using indirect fluorescence detection. AB - A method of using indirect fluorescence detection is evaluated for the analysis of glucosamine in commercial dietary supplements following chromatographic separation. In this method, the eluting analyte, glucosamine, was detected by monitoring an increase in the fluorescence signal for L-tryptophan (L-Trp) or DL 5-methoxytryptophan (5-MTP) after glucosamine complexed with a copper(II) ion and released either L-Trp or 5-MTP from a copper(II) complex, which is introduced postcolumn. The fluorescence of L-Trp and 5-MTP are quenched when complexed with the copper(II) ion. The results obtained using indirect fluorescence detection are compared with the results obtained for precolumn derivatization with phenylisothiocyanate. Statistical analysis is performed to compare the results obtained for the two postcolumn interaction components, Cu(L-Trp)2 and Cu(5 MTP)2, as well as the results obtained using the indirect fluorescence detection method and a precolumn derivatization method. The indirect fluorescence detection method provided an alternative to precolumn derivatization for determining the concentration of glucosamine in commercial dietary supplements. The stability of the glucosamine-o-phthalaldehyde-3-mercaptopropionic acid derivative is also evaluated to investigate the applicability of the popular precolumn derivatization reagent, o-phthalaldehyde-3-mercaptopropionic acid, for this analysis. PMID- 17425136 TI - Preparative-scale chromatography of ecdysteroids of Serratula wolffli andrae. AB - Numerous ecdysteroids are isolated from the herb of Serratula wolffii Andrae, a cultivated plant. The isolation procedure includes a variety of low-pressure liquid chromatography, thin-layer chromatography (TLC), gel chromatography, and high-performance liquid chromatography (HPLC) methods. The progress of separation is monitored by TLC, and the final proof of purity is carried out by HPLC. The isolation process involves the removal of proteins, flavonoids, chlorophylls, other sterines, etc. The purification also includes the separation of the target ecdysteroids from each other. Isolation of the pure compounds requires 2-8 chromatographic steps. The consecutive steps are based on the different physicochemical properties of the ecdysteroids. In some cases, a special peak-cut method employing a flush of dichloromethane into the dichloromethane-isopropanol water mobile phase is used. This flush of dichloromethane leads to an almost perfect separation of otherwise unresolved peaks. Two ecdysteroids, 25 hydroxydacryhainansterone and 14-epi-20-hydroxyecdysone, are identified as natural products for the first time. The structure-chiroptical relationships for some ecdysteroids are also discussed. PMID- 17425137 TI - Quantitative determination of valienamine and validamine by thin-layer chromatography. AB - A simple and valid thin-layer chromatographic method for the separation and quantitative determination of valienamine and validamine is described. The two compounds are separated using a Silica gel G plate as the stationary phase and a mixture of 1-PrOH-AcOH-H2O (4:1:1, v/v/v) as the mobile phase. The plate is developed for 1 h at 25 degrees C and dried by a hairdrier, then immersed in 0.1% ninhydrin aqueous solution and heated for 5 min at 121 degrees C. The reacted spots are scanned with a single wavelength at 420 nm in the measurement mode of absorption. The limits of detection of the two compounds are both 0.4 microg. The responses of the densitometry are highly correlated with the amounts of valienamine and validamine in the range of 0.4-2.8 pg. Moreover, the method shows good accuracy and high precision. PMID- 17425138 TI - Rapid and sensitive method for the determination of sibutramine active metabolites in human plasma by reversed-phase liquid chromatography-tandem mass spectroscopy. AB - A new, rapid, and sensitive liquid chromatography-tandem mass spectrometry method is developed and validated to quantitate the sibutramine active metabolites mono desmethyl sibutramine (M1) and di-desmethyl sibutramine (M2) using imipramine as the internal standard in human plasma samples for routine bioequivalence studies. The method involves rapid solid-phase extraction from plasma, eliminating the drying and reconstitution steps. The analytes are chromatographed on a C8 reversed-phase chromatographic column and analyzed by mass spectrometry in the multiple reaction monitoring mode, which enables a quantitation limit at the sub nanogram level. The method has a chromatographic run time of 2.8 min. The proposed method is validated with a linear range of 0.1-8.0 and 0.2-16.0 ng/mL for M1 and M2, respectively, with a correlation coefficient of regression > or = 0.9990. The method is sensitive and reproducible, having intra- and inter-assay precision at the lower limit of quantitation (0.1 ng/mL for M1 and 0.2 ng/mL for M2) < 10.0%. The overall recovery for M1 and M2 is 93.5% and 77.9%, respectively. The method has been applied to a bioequivalence clinical study with great success. PMID- 17425139 TI - Quantitative determination of galantamine in human plasma by sensitive liquid chromatography-tandem mass spectrometry using loratadine as an internal standard. AB - A simple, rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry method is developed and validated for the quantitation of galantamine, an acetylcholinesterase inhibitor in human plasma, using a commercially available compound, loratadine, as the internal standard. Following liquid-liquid extraction, the analytes are separated using an isocratic mobile phase on a reverse-phase C18 column and analyzed by mass spectrometry in the multiple reaction monitoring mode using the respective (M+H)+ ions, m/z 288 to 213 for galantamine and m/z 383 and 337 for the internal standard. The assay exhibit a linear dynamic range of 0.5-100 ng/mL for galantamine in human plasma. The lower limit of quantitation is 0.5 ng/mL, with a relative standard deviation of less than 8%. Acceptable precision and accuracy are obtained for concentrations over the standard curve range. A run time of 2.5 min for each sample makes it possible to analyze more than 400 human plasma samples per day. The validated method is successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability, or bioequivalence studies. PMID- 17425140 TI - Separation of polar mushroom toxins by mixed-mode hydrophilic and ionic interaction liquid chromatography-electrospray ionization-mass spectrometry. AB - Reversed-phase liquid chromatography (RPLC) is commonly used to analyze nonvolatile contaminants and naturally occurring toxins in foods. However, polar compounds, such as hydrophilic polypeptides and quaternary ammonium salts, are often not satisfactorily separated by RPLC and present a challenge for analytical scientists. In this study, hydrophilic interaction liquid chromatography (HILIC), on an amide-based stationary phase in combination with electrospray ionization (ESI) tandem mass spectrometry (MS-MS), is successfully employed to simultaneously separate polar mushroom toxins, including amanitins and phallotoxins, which are cyclic oligopeptides and muscarine, a quaternary ammonium compound, in mushrooms. The sensitivity of different ionization modes is studied, and the positive ionization mode is found to provide a more sensitive and effective tool for the unambiguous identification of the concerned polar toxins because of their characteristic fragmentation patterns. The properties of the mobile phase are also found to have significant impacts on the separation. At a high acetonitrile (ACN) concentration, hydrophilic interaction dominates, and all analytes under study demonstrate a much higher affinity with the stationary phase. The addition of methanol (MeOH) as a modifier could further enhance the HILIC separation for amanitins, phallotoxins, and muscarine. Valley-to-valley separation is achieved upon the optimatizatiqn of the mobile phase (comprising of ACN, MeOH, and ammonium formate buffer at pH approximately 3.5) and the solvent gradient. HILIC coupled with ESI-MS-MS is demonstrated to be a novel technique for the simultaneous separation and confirmatory analysis of the concerned polar toxins by providing an environment of solubility and retention that could not be achieved through the use of RPLC. PMID- 17425141 TI - Gas chromatography problem solving and troubleshooting. Why is the Vespel material recommended? PMID- 17425142 TI - Parrhesia as a conceptual metaphor for nursing advocacy. PMID- 17425143 TI - Moral responsibility: a relational way of being. AB - This article reports a study exploring the meaning of the complex phenomenon of moral responsibility in nursing practice. Each of three focus groups with a total of 14 student nurses were conducted twice to gather their views on moral responsibility in nursing practice. The data were analysed by qualitative thematic content analysis. Moral responsibility was interpreted as a relational way of being, which involved guidance by one's inner compass composed of ideals, values and knowledge that translate into a striving to do good. It was concluded that, if student nurses are to continue striving to do good in a way that respects themselves and other people, it is important that they do not feel forced to compromise their values. Instead they should be given space and encouragement in their endeavours to do good in a relational way that advances nursing as a moral practice. PMID- 17425144 TI - Ethical sensitivity: state of knowledge and needs for further research. AB - Ethical sensitivity was introduced to caring science to describe the first component of decision making in professional practice; that is, recognizing and interpreting the ethical dimension of a care situation. It has since been conceptualized in various ways by scholars of professional disciplines. While all have agreed that ethical sensitivity is vital to practice, there has been no consensus regarding its definition, its characteristics, the conditions needed for it to occur, or the outcomes to professionals and society. The purpose of this article is to explore the meaning of the concept of ethical sensitivity based on a review of the professional literature of selected disciplines. Qualitative content analysis of the many descriptors found within the literature was conducted to enhance understanding of the concept and identify its essential characteristics. Ethical sensitivity is considered to be an emerging concept with potential utility in research and practice. PMID- 17425145 TI - Dutch nurses' views on codes of ethics. AB - This study explored the experiences and views of Dutch nurses on the content, function, dissemination and implementation of their codes of ethics. A total of 39 participants, who differed in age, qualifications, length of work experience and health care setting, took part in focus groups. The findings revealed common unfamiliarity with and a rather implicit use of codes, and negative comments on the growing number of codes available in the Netherlands. Limited dissemination, implementation and functioning of codes of ethics were also identified. The findings were discussed using concepts from the literature, nursing practice and personal experience. PMID- 17425146 TI - Moral obligations of nurses based on the ICN, UK, Irish and Polish codes of ethics for nurses. AB - A code of professional conduct is a collection of norms appropriate for the nursing profession and should be the point of reference for all decisions made during the care process. Codes of ethics for nurses are formulated by members of national nurses' organizations. These codes can be considered to specify general norms that function in the relevant society, adjusting them to the character of the profession and enriching them with rules signifying the essence of nursing professionalism. The aim of this article is to present a comparative analysis of codes of ethics for nurses: the ICN's Code of ethics for nurses, the U.K.'s Code of professional conduct, the Irish Code of professional conduct for each nurse and midwife, and the Polish Code of professional ethics for nurses and midwives. This analysis allows the identification of common elements in the professional ethics of nurses in these countries. PMID- 17425147 TI - Development of the perceptions of conscience questionnaire. AB - Health care often involves ethically difficult situations that may disquiet the conscience. The purpose of this study was to develop a questionnaire for identifying various perceptions of conscience within a framework based on the literature and on explorative interviews about perceptions of conscience (Perceptions of Conscience Questionnaire). The questionnaire was tested on a sample of 444 registered nurses, enrolled nurses, nurses' assistants and physicians. The data were analysed using principal component analysis to explore possible dimensions of perceptions of conscience. The results showed six dimensions, found also in theory and empirical health care studies. Conscience was perceived as authority, a warning signal, demanding sensitivity, an asset, a burden and depending on culture. We conclude that the Perceptions of Conscience Questionnaire is valid for assessing some perceptions of conscience relevant to health care providers. PMID- 17425148 TI - Whistleblowing in Japan. AB - This article, written from research data, focuses on the possible meaning of the data rather than on detailed statistical reporting. It defines whistleblowing as an act of the international nursing ethical ideal of advocacy, and places it in the larger context of professional responsibility. The experiences, actions, and ethical positions of 24 Japanese nurses regarding whistleblowing or reporting a colleague for wrongdoing provide the data. Of these respondents, similar in age, educational level and clinical experience, 10 had previously reported another nurse and 12 had reported a physician for a wrongful act. These data raise questions about overt actions to expose a colleague in a culture that values group loyalty and saving face. Additional research is needed for an in-depth understanding of whistleblowing, patient advocacy and professional responsibility across cultures, especially those that value group loyalty, saving face and similar concepts to the Japanese Ishin Denshin, where the value is on implicit understanding requiring indirect communication. Usually, being direct and openly discussing sensitive topics is not valued in Japan because such behavior disrupts the most fundamental value, harmony (wa). PMID- 17425149 TI - Professional ethics as an important factor in clinical competency in nursing. AB - It is imperative to understand the factors that influence clinical competency. Consequently, it is essential to study those that have an impact on the process of attaining clinical competency. A grounded theory approach was adopted for this study. Professional competency empowers nurses and enables them to fulfill their duties effectively. Internal and external factors were identified as affecting clinical competency. A total of 36 clinical nurses, nurse educators, hospital managers and members of the Nursing Council in Tehran participated in this research. Data were obtained by semistructured interviews. Personal factors and useful work experience were considered to be significant, based on knowledge and skills, ethical conduct, professional commitment, self-respect and respect for others, as well as from effective relationships, interest, responsibility and accountability. Effective management, education systems and technology were named as influential environmental factors. Personal and environmental factors affect clinical competency. Ethical persons are responsible and committed to their work, acquiring relevant work experience. A suitable work environment that is structured and ordered also encourages an ethical approach by nurses. PMID- 17425150 TI - Human dignity in paediatrics: the effects of health care. AB - Human dignity is grounded in basic human attributes such as life and self respect. When people cannot stand up for themselves they may lose their dignity towards themselves and others. The aim of this study was to elucidate if dignity remains intact for family members during care procedures in a children's hospital. A qualitative approach was adopted, using open non-participation observation. The findings indicate that dignity remains intact in family-centred care where all concerned parties encourage each other in a collaborative relationship. Dignity is shattered when practitioners care from their own perspective without seeing the individual in front of them. When there is a break in care, family members can restore their dignity because the interruption helps them to master their emotions. Family members' dignity is shattered and remains damaged when they are emotionally overwhelmed; they surrender themselves to practitioners' care, losing their self-esteem and self-respect. PMID- 17425151 TI - Supporting stroke patients' autonomy during rehabilitation. AB - In a qualitative study, 22 stroke patients undergoing rehabilitation in three nursing homes were interviewed about constraints on and improvements in their autonomy and about approaches of health professionals regarding autonomy. The data were analysed using grounded theory, with a particular focus on the process of regaining autonomy. An approach by the health professionals that was responsive to changes in the patients' autonomy was found to be helpful for restoration of their autonomy. Two patterns in health professionals' approach appeared to be facilitatory: (1) from full support on admission through moderate support and supervision, to reduced supervision at discharge; and (2) from paternalism on admission through partial paternalism (regarding treatment) to shared decision making at discharge. The approach experienced by the patients did not always match their desires regarding their autonomy. Support and supervision were reduced over time, but paternalism was often continued too long. Additionally, the patients experienced a lack of information. Tailoring interventions to patients' progress in autonomy would stimulate their active participation in rehabilitation and in decision making, and would improve patients' preparation for autonomous living after discharge. PMID- 17425152 TI - Ethical problems in end-of-life decisions for elderly Norwegians. AB - Norwegian health professionals, elderly people and family members experience ethical problems involving end-of-life decision making for elders in the context of the values of Norwegian society. This study used ethical inquiry and qualitative methodology to conduct and analyze interviews carried out with 25 health professionals, six elderly people and five family members about the ethical problems they encountered in end-of-life decision making in Norway. All three participant groups experienced ethical problems involving the adequacy of health care for elderly Norwegians. Older people were concerned about being a burden to their families at the end of their life. However, health professionals wished to protect families from the burden of difficult decisions regarding health care for elderly parents at the end of life. Strategies are suggested for dialogue about end-of-life decisions and the integration of palliative care approaches into health care services for frail elderly people. PMID- 17425153 TI - 'The globalisation of nursing: ethical, legal and political issues' university of surrey 10-11 july 2006: a summary of the deliberations of the concurrent working groups. PMID- 17425154 TI - An ethical voice for nurses--is anybody listening? PMID- 17425155 TI - Mitochondria and mitochondrial nitric oxide synthase alterations participate in energetical dysbalance, aging and age-related diseases. AB - Nitric oxide (NO) is a unique mediator of cellular regulations synthesized by nitric oxide synthases (NOS) present in cytoplasm of various cells. An additional Ca-dependent mitochondrial NOS (mtNOS) detected just recently synthesizes also NO inhibiting oxidative phosphorylation, i.e. mitochondrial energy producing metabolic process and protects mitochondria from oxygen radicals. Mitochondrial membrane possesses electrogenic uniporter transporting Ca into mitochondria (stimulation of mtNOS), while Na+/Ca2+ exchanger removes Ca from mitochondria. Mitochondrial disorders with low mtNOS activity participate in accelerated aging and age-related diseases. The direct NO balance determination is outside of the standard clinical facilities; Indirect alternatives, such as insulin resistance determination are accessible. Pharmacotherapy exploits effective therapeutic and preventive measures (NO donors, ACEI inhibitors, etc.) and pharmaceutical approach (development of mitochondriotropic drugs). We suggest, that mitochondrial disorders participate in aging and age related diseases and propose that the early diagnostics, preventive and therapeutic measures could prevent and even correct at least partially the development of age-related diseases (Tab. 4, Ref. 81). PMID- 17425156 TI - Avian influenza--a new challenge. AB - OBJECTIVE: To present a wide-ranging review of the literature on avian influenza A (H5N1). BACKGROUND: The recent epidemics caused by the avian influenza A virus in Asia, have demonstrated the capacity of this agent to cause serious illness in humans. METHODS: Most articles were obtained from the Medline database using the keywords "Influenza A virus", "avian flu", "epidemiology", "disease vectors" and "H5N1 infection" for the period between 1996 and 2006. RESULTS: We selected 25 original articles addressing the recent outbreaks of infection with the H5N1 subtype of avian influenza A in domesticated birds in Asia, which have resulted in significant economic losses and repercussions for public health, as well as some cases of human infection presenting high lethality. In most cases, infection has been associated with direct exposure to infected birds or contact with surfaces infected with bird excrement. However, cases of human-to-human transmission have been confirmed. CONCLUSION: Controlling outbreaks in domestic fowl and limiting contact between humans and infected birds must be the priorities in the management of this disease at the public health level. In addition, techniques and knowledge regarding the disease should be widely disseminated (Ref: 28). PMID- 17425157 TI - Systematic isolation and characterization of cDNAs encoding AAA proteins from human brain. AB - BACKGROUND: The AAA (ATPases Associated with various cellular Activities) domain characterizes a diverse superfamily of proteins. Mutations in genes encoding AAA domains cause a variety of human diseases including cystic fibrosis, Zellweger syndrome, adrenomyeloneuropathy, and dystonia. Recently, mutations in two AAA containing proteins paraplegin and spastin have been shown to cause two types of hereditary spastic paraplegia (HSP). The HSPs are genetically heterogeneous degenerative spinal cord disorders characterized by lower extremity weakness and spasticity. Clinical similarity between various genetic types of HSP led us to propose that different genetic types of HSP were due to common biochemical abnormalities including disturbances in related proteins. For this reason, we sought to identify novel AAA-containing proteins as potential candidates for HSP and related neurodegnerative disorders. We used degenerative PCR, based on the conserved AAA peptide sequence to systematically clone and characterize AAA genes expressed in human brain. RESULTS: We analyzed 646 clones and identified 19 known AAA-containing proteins including spastin and paraplegin, AAA-containing genes that cause HSP. In addition, we identified 14 unique DNA inserts representing novel putative AAA-containing proteins. Four of these novel genes are hypothetical AAA proteins and the rest of novel clones matched sequences of yet uncharacterized expressed sequence tags (ESTs). CONCLUSION: Fourteen novel AAA containing proteins are potential candidates for human diseases including degenerative neurologic disorders, and their further analysis is ongoing (Tab. 1, Fig. 1, Ref. 22). PMID- 17425158 TI - Correlation between the aPTT and clinical events in acute coronary syndromes treated with unfractionated heparin. AB - OBJECTIVE: The primary objective of this study is to analyse relationship between activated partial thromboplastin time (aPTT) and clinical events in patients with unstable coronary artery disease. BACKGROUND: Unfractionated heparin (UFH) is considered as a standard therapy in the management of unstable coronary artery disease. His efficacy is concerned to low rate of adverse events, but it shows lack of correlation between aPTT and clinical outcomes. METHODS: Thirty patients with acute coronary syndromes were randomised to receive weight-adjusted unfractionated heparin (Heparin-"Biochemie" 25.000 IU/5 ml), according to Rashke nomogram. Six hours after the initial bolus dose, the aPTT was measured and the heparin infusion was adjusted. Relationship between the aPTT and the day of onset of recurrent angina (RA), myocardial infarction (MI), heart failure (HF), death, and target vascular revasularization (TVR) during six months follow-up was evaluated. RESULTS: At 6 h, 40 % of patients achieved the target level of aPTT (60-85 sec). Also, 40 % patients were below the target range, 41.6 % had aPTT less than 30 sec and 20 % were above the target range. We noted significant difference between heparin dosing and aPTT (p=0.0000). But we did not find significant difference between aPTT and outcomes as RA and MI (p=1.0, p=0.42). Pearson Product Moment Correlation shows negative correlations between aPTT and the day of onset of RA (r=-0.05) and TVR (r=-0.23), and pozitive correlations for MI (r=0.35) and death (r=0.87). CONCLUSION: The level of aPTT shows discordance with the onset of clinical events and is a weak predictor of outcomes (Tab. 2, Fig. 7, Ref 5). PMID- 17425159 TI - Valsalva leak point pressure predicts outcome after transobturator suburethral tape implantation--fact or fiction? AB - INTRODUCTION AND OBJECTIVES: The aim of the study was to assess the influence of Valsalva leak point pressure (VLPP) on outcome of transobturator suburethral tape (TOT) implantation. MATERIAL AND METHODS: 65 female patients underwent surgery between March 2004 and November 2005. Their inclusion criterion was urodynamic stress urinary incontinence (SUI). The exclusion criteria were detrusor overactivity, detrusor underactivity, a significant post-void residuum and genital prolapse. "Subjective improvement" was considered as the difference between postoperative urinary-incontinence specific quality of life compered with its preoperative value (deltaIQoL=IQoL*-IQoL). RESULTS: Postoperative leakage and postoperative urinary incontinence specific quality of life in 1st versus 2nd group were 1.2+/-1.1 g and 90.6+/-5.1 versus 4.6+/-3.1 g and 84.0=5.6 respectively (p<0.005 and p<0.005 respectively). CONCLUSIONS: Cure and success rates were significantly higher in patients with higher VLPP values compared with patients with lower VLPP values. In spite of the fact that patients with higher VLPP values achieved significantly lower postoperative leakage, urine leakage decreased significantly more in patients with lower VLPP values. Patients with higher VLPP values reached significantly higher postoperative urinary incontinence-specific quality of life, however no significant difference was found in subjective improvement between patients with higher and lower VLPP values (Tab. 1, Ref. 23). PMID- 17425160 TI - Long-term results of surgical treatment of patients suffering from cerebral palsy. AB - OBJECTIVES: Cerebral palsy is defined as a syndrome subsequent to attack of developing brain and characterized by loss of normal controle of motor function. Orthopaedic surgeon have to select patients who will have benefit from surgery, and to perform it by that way, that will improve the motorical ability of the child. BACKGROUND: The operations performed in the past at one level were replaced by modern ,,multilevel approach" in case of need. The aim of this study was the analysis of the long term results of that new surgical approaches. METHODS: Evaluated were 275 patients diagnosed as having spastic or mixed form of cerebral palsy (CP). The average age at the time of surgery was 7.8 years (3 to 16 years). Altogether in 20 years 941 surgical procedures, mainly on the lower limbs were performed. RESULTS: Overall 9 % excellent, 53 % very good, 26 % good and 12 % bad results were achieved. Using single level unilateral operations 7 % excellent, 46 % very good, 30 % good and 17 % fair results were achieved. Comparing to this, using the modern methods, 11 % excelent, 57 % very good, 23 % good and 9 % of bad results were achieved. CONCLUSION: Authors believe, that the broad and early multilevel surgery, acomplished with proper physiotherapy is an important fact for improvement of outcome in treatment of children suffering from cerebral palsy (Tab. 2, Ref 6). PMID- 17425161 TI - Is interhemispheric arachnoid cyst with corpus callosum agenesis related to cavum vergae cyst? AB - Authors operated on successfully giant interhemispheric arachnoid cyst associated with corpus callosum agenesis. A conclusion was drawn that this cyst was in fact expansive cavum Vergae cyst, taking into consideration the developmental mechanism responsible for the formation of third ventricle dorsal cysts (Fig. 8, Ref. 10). PMID- 17425162 TI - Retropharyngeal lymphangioma. AB - Retropharyngeal lymphangioma is exceedingly rare and has been reported in only one case report in English literature. We are presenting a case of a 21 month-old girl with a history of acute tonsillitis and acute inspiratory stridor due to retropharyngeal bulging. A needle aspiration was done for decompression of the airway and suspected retropharyngeal abscess. Aspiration produced transparent fluid and a CT scan with contrast excluded abscess formation. Because the lymphangioma persisted in spite of two repeated aspirations and with continued presence of inflammation, we used a transcervical approach to excise the cervical lymphangioma. After surgery dehydrated alcohol was injected into the retropharyngeal space. 27 months later there was no sign of recurrence (Fig. 2, Ref 12). PMID- 17425163 TI - Maggot debridement therapy. AB - AIM: To establish a maggot debridement therapy unit and assess efficacy of this method in chronic conservative non-treatable leg ulcers in patient in whom conventional therapy failed. MATERIAL AND METHODS: From August 2004 to December 2005 we treated with Maggot debridement therapy (MDT) 10 patients, 6 men and 4 women, with 13 leg ulcers treated first with conventional and then with maggot therapy. Average age of these patients was 55 years, 11 wounds were of venous and 2 of arterial origin. 7 patients had diabetes mellitus. RESULTS: 1 patient with arterial ulcers underwent supracondylar amputation, 2 patients were re-admitted for recurrent ulcers, one extremely obese patient with cardiovascular co morbidity did not heal, but all other ulcers have been healed, cleared or minimised during 4-8 week MDT. We observed massive growth of granulation tissue and microbiological cleaning of these wounds. No adverse effects were observed. CONCLUSION: Maggot therapy was more effective and efficient in debriding non healing leg ulcers than a conservative treatment (Fig. 3, Ref. 6). PMID- 17425164 TI - Giant solid abdominal mass with cystic lesions: a case report and diaphorodiagnostic approach. AB - A peri-menopausal woman presented with abdominal distention, pelvic discomfort and problems of constipation for the last 4 months. All clinical and radiological examinations were in favor of a giant solid mass with cystic lesions arised from the left ovary. These findings raised suspicion of a primary malignant ovarian tumor or a preudomyxoma peritonei. Surgery revealed a giant mass arised from the uterine fundus. An abdominal hysterectomy with bilateral salpingo-oophorectomy and omentectomy were preformed. The histological examination verified a degenerated myoma with cystic lesions with no evidence of malignancy. The patient made an uneventful recovery. A gynecologist should always be prepared to perform a different surgery than planned according to operational findings (Fig. 2, Ref. 11). PMID- 17425165 TI - Coagulase negative staphylococci and their participation in pathogenesis of human infections. AB - Coagulase negative staphylococci are among the bacteria routinely isolated at various clinical microbiology departments. There are presently 41 recognized taxons, designated coagulase negative staphylococci. Although long considered nonpathogens as the components of normal human skin and mucosa, lately they turned into significant etiological agents causing nosocomial infections, mainly in link with the presence of foreign bodies in the human organism. In addition, their increasing resistance to antibacterial drugs evoked deepening concern of infections involving coagulase negative staphylococci. Problems already appear during interception of the coagulase negative staphylococci in the material and the subsequent determination, whether they are invaders or colonisers. Hence, major efforts have been made toward the development of methods enabling to recognize those properties of coagulase negative staphylococci, which would resolve this day-to-day dilemma, facing the clinical microbiologist. From the aspect of their pathogenic potential, the most important property of coagulase negative staphylococci is their capability to form biofilm on the surfaces of foreign bodies introduced (implanted) into the organism. In consequence of such formation they are protected in the biofilm against the effects of antibacterial drugs and of the immune system. Lately, major interest appeared in the study of factors, mainly S. epidermidis, which participate in the biofilm formation, as well as of the determination of virulence factors of other coagulase negative staphylococci (Tab. 2, Ref. 25). PMID- 17425166 TI - Which statistical tests for estimating osteoporotic fracture risk? AB - The article presents an analysis of statistical methods used for estimating fracture risk in patients with osteoporosis. Mathematical relations of different methods are explained (risk--R, risk ratio--RR, RD--risk difference, odds--O, odds ratio--OR, Yule's Q, Yule's Y, logistic model). What is important to keep in mind is that: 1) relative risk and odds ratio are statistics that only describe an association, not causation; 2) relative risk and odds ratio refer to a population, not to an individual patient; 3) the studies of small groups are more likely to find an association that might actually just be due to chance, the larger the groups, the less likely the association between a risk factor and an outcome (fracture); 4) when the incidence of an outcome of interest in the study population is low (<10 %), the OR is close to the RR, the more frequent the outcome becomes, the more the OR will overestimate the RR when it is more than 1 or underestimate the RR when it is less than 1. Sophisticated statistical packages are available which can calculate many of the tests of association but the problem is that the investigator must know which the desirable is. The incorrect option of statistical analysis, the incorrect interpretation of risk ratio or odds ratio and overestimation of the importance of a risk factor may lead to unintentional errors in the economic analysis of potential programs or treatments in osteoporosis. This article could be a contribution for investigators, who are concerned with assessment of fracture risk. PMID- 17425167 TI - University publication database. AB - Information technology plays an important role in the strategic initiatives of higher education institutions. Information systems are used to support accreditation efforts, integrate curriculum, solve administrative information problems, etc. The article briefly describes the information needs of higher education institutions, specifically the needs for data on publication activities (Fig. 3, Ref. 1). PMID- 17425168 TI - [Anorexia nervosa]]. PMID- 17425169 TI - [Environmental health. Medical aspects--children's unique vulnerability- perception of the environment]. AB - It is recognized that humans are participants in complex ecosystems and that their potential for health is proportional to the health function of those ecosystems ("ecosystem health"). From the medical viewpoint this close connection between environment and health is outlined acknowledging that man-made environmental degradation may lead to human illness. This is the very object of the "environmental medicine" which should also take into account psycho philosophical aspects related to the affective perception of our environment and its degradation. Data are reviewed on the special vulnerability of children as an example of groups within the population who are particularly sensitive to toxic hazards in the environment. The vulnerability of infants and children reflects the unique patterns of exposure to environmental hazards, coupled with the inherent fragility of their developmental processes. Given this example, the concept is explored that certain groups within our population, most notably children, are deserving of special protection in risk assessment, law, and regulation. As physicians, we should use prudence when counselling our patients, especially pregnant mothers, about avoidance of exposures to chemicals of unknown and untested toxic potential. PMID- 17425170 TI - [Vaccination in immunocompromised patients]. AB - Vaccination strategies in immunocompromised patients differ from those in immunocompetent patients: The risk of severe and complicated infections is increased, the immune response to vaccination might be hampered and side-effects- especially vaccination-induced infections after live-vaccines--must carefully be weighed against benefit of vaccination. Encouragement of vaccination, adequate timing, boostering and newer immunological strategies can improve immunity. This article summarizes vaccination problems in patients with HIV/AIDS, Solid-organ transplantation, haematological stem cell transplantation, splenectomy and immunosuppressive drugs. PMID- 17425171 TI - [Everybody needs an X-chromosome]. PMID- 17425172 TI - [Tick borne encephalitis 2006]. PMID- 17425173 TI - [Gifts and payments: notes on the history and psychology of medical fees]. PMID- 17425174 TI - ["Injured at rescue, moribund during transport, dead at clinic admission"--trauma rescue work and medical intervention in Germany 1950-1970]. PMID- 17425175 TI - [Egg: concepts, analyses and controversies in the human health]. AB - Cholesterol rich-foods consumption has been related as a causing factor for heart ischemic disease. Because the high cholesterol content of the yolk, the egg consumption has been decreased in spite of it, egg is a complete food, rich in many nutrients, and economically accessible. The cholesterol content in egg yolk has also important properties for the human organism. The present work had the objective to review the scientific literature about egg's cholesterol, describing the possible consequences on the human health and wellbeing, its effect when they are enriched, and the chemical composition in relation to its lipidic profile. Information for this review was collected through national and international inquiries. According to this information, most of the studies on egg consumption are not related to the risk of cardiopathies in healthy people. Also, in people with diabetes mellitus, there is scarce evidence to drawn any conclusion about egg consumption and cardiopathies. Omega-3 egg enrichment presumably possesses a protective effect against cancer, despite its cholesterol content remains unaltered. Many of the Food Chemical Composition Tables contain different values on egg's fat composition, so it should be continuously update to reduce these discrepancies. Also, more studies on omega-3 enriched eggs are necessary for deeper conclusions on their cardio-protective effect. PMID- 17425176 TI - [Secular trends in growth in male adolescents: height and weight gains, nutritional state and relation with education]. AB - This work aimed at characterizing the modifications in adolescents nutritional status, growth evolution and education along time and the correlation with the height, weight and body mass index. Data came from 2616 male adolescents soldiers between 17 and 19 years old enlisted for Armed Services in the city of Vicosa Minas Gerais, Brazil, between 1995 and 2004. There was a positive secular trend for height and weight, with a 4cm and 3kg median increment, respectively, along time. There was a reduction from 28.0% to 11.6% on the prevalence of short stature in the period. It was estimated positive correlation (p < 0.05) between education and stature, weight and BMI. The prevalence of weight excess (risk of overweight and overweight) increased from 7.1% (1995) to 9.1% (2004). Although it was observed positive secular trend for stature and weight, it was not sufficient to reach the median of the National Center for Health Statistics/Center for Disease Control and Prevention. Weight excess and short stature prevalence must be monitored, being necessary the implementation of measures that focus to prevent these disturbance and aim at reaching or maintaining an adequate nutritional state for future generations. PMID- 17425177 TI - [Serum levels of interferon-gamma and interleukine-10 in anemic children with vitamin A deficiency]. AB - Studies done with human and animals have shown that the diminished vitamin A intake can produce anemia, but the ethiopathogenic mechanism hasn't been known due to the few studies about the complex interactions between micronutrients deficiencies, immune system and cytokine productions. The objective of the present study was to analyze the serum concentration of IFN-gamma and IL-10 in anemic and non anemic children according to vitamin A nutritional status. A descriptive study of 53 euthrophic children, without infectious diseases (45-77 month old; 29 girls, 24 boys), living in a slum in Maracaibo-Venezuela. Anemia was evaluated by means of blood hemoglobin concentration analysis, standard WHO criteria for anemia were Hb < 110 g/L in <60 month old children; Hb < 115 g/L in >60 month old children. Vitamin A status was assessed by Conjunctival Impression Cytology (CIC) technique, which determines Normal CIC=Control or Abnormal CIC=VAD. The serum concentration of IFN-gamma and IL-10 (pg/mL; X +/- DE), was determined by the ELISA method. Statistical significance of differences was tested by ANOVA (p < 0.05). Prevalence of anemia was 75.50%; VAD 50.94% and anemia+VAD 35.86%. The anemic children with VAD showed significant serum values of IFN-gamma e IL-10 lower than in other groups. This alteration of the complex immunoregulatory mechanism could be responsible for dysfunction eritrhoid cell formation. PMID- 17425178 TI - [Serum total homocysteine and lipoproteins levels in young adults from urban and rural areas of Costa Rica]. AB - Serum total homocysteine and lipoproteins levels in young adults from urban and rural areas of Costa Rica. This study evaluated the lipid profile and the serum levels of total homocysteine of 400 adults aged 20 to 40 years, residents of rural and urban areas of the central valley of Costa Rica. The results showed that the proportion of subjects from urban areas with high levels of LDL-C (= 4.13 mmol/L) was 11% higher than the proportion of adults from rural areas (29.2% and 18.0%, p = 0.045). On the contrary, the proportion of rural subjects with low HDL-C levels (< 1.03 mmol/ L) was higher 19% than the proportion of urban adults (56.0% and 75.3%, p = 0.001). Rural residents also showed higher serum levels of triglycerides and total homocysteine than the urban residents, although the differences were not significant (TG: 1.94 mmol/L and 2.02 mmol/L, p = 0.529; TH: 9.33 micromol/L and 9.84 micromol/L, p = 0.145). The prevalences of hyperhomocysteinemia (>15 micromol/L), hypercholesterolemia (= 5.17 mmol/L) and hypertriglyceridemia (= 1.69 mmol/L) in the studied population were 5.8%, 53.6% and 50.1% respectively. 82% of the studied population had LDL-C levels above 2.58 mmol/L and approximately 61% of the adults had HDL-C levels lower than 1.03 mmol/L. Our data suggest that the studied costarrican population has a very high cardiovascular risk. The prevalence of low levels of HDL-C (< 1.03 mmol/L), marginal levels of total homocysteine and a tendency to have high levels of triglycerides were the most outstanding cardiovascular risk factors found in this population, specially in rural subjects, which could be associated with the quality of the diet of these habitants. It is necessary to develop effective intervention strategies to promote healthy lifestyles in the population in order to reduce the cardiovascular mortality rates in Costa Rica. PMID- 17425179 TI - [Helicobacter pylori infection (13C-UBT), and its relationship with nutritional and socioeconomic factors in low income school children from Valencia, Venezuela]. AB - Helicobacter pylori infection (Hp) is widely spread around the world, and it is considered one of the main causes of chronic gastritis, peptic and duodenal ulcers, and gastric cancer. Recent research has shown that it can be associated with nutritional disorders, mainly with iron and other micronutrient deficiencies. The objective of this study was to assess the prevalence of Hp infection, and infection pattern according to age, sex, nutritional status, and socioeconomic conditions in children who attended the Unidad Educativa "Valentin Espinal" in the city of Valencia. 170 children, between 3 and 14 years of age were studied to assess Hpylori infection (13C-urea breath test), age, nutritional status according to BMI and Height for age, hemoglobin (cianometahemoglobin), serum ferritin (ELISA), socioeconomic status (Graffar-Mendez-Castellano), housing conditions, number of families and of people cohabitating in the same household, and quality of services. 78.8% of the children were infected with Hp, witch was significantly correlated with age but not gender. 25.9% of the sample had undernutrition, and 46.5% were stunted. 98.1% of the families lived in poverty, and 98% of the households showed sanitary deficiencies. A mean of 6.0 +/- 2.4 persons lived in each household (range: 2-15), and an average of 3.2 person shared bedrooms. The odds of being infected were higher in those children who were stunted. Also, socioeconomic status, mother's education level, and poor hosing conditions were significantly associated to being infected. Hpylori is highly prevalent among socially and economically deprived children, and age, overcrowding, and a low education level of the mother increases the risk of being infected. PMID- 17425180 TI - [Study of the reineta protein modifications (Brama australis), put under freezing and storage to -18 degrees C and -30 degrees C]. AB - The objective of the present work was to study functional and thermal properties of reineta (Brama australis) frozen meat, analysed by water retention capacity (WRC), gel forming capacity (GFC), texture, emulsifying capacity and differential scanning calorimetry (DSC). For this study, reineta fillets were obtained and extracted by the same conditions, and cutted, packaged, frozen and stored at -18 degrees C and -30 degrees C for 7 months. The results obtained, showed that there were no signifficant differences in the responses to thermal treatment for all the specimens. For samples frozen at -18 degrees C and -30 degrees C, the protein contents were 23.5 + 0.0 and 25.4 + 1.0%, respectively. The WRC values were 0.45 + 0.1 and 1.59 +/- 0.0 g water/g protein, respectively. The gel forming capacity was only present in the fresh samples, whereas the frozen stored ones only form protein aggregates. The emulsifying capacity was between 960 and 1400 g oil / g protein, and the storage time increased this value. The miosin denaturation temperature (Td) and denaturation enthalpy (?H), obtained by DSC, fluctuated between 39.2 +/- 0.5 to 44.8 +/- 0.8 degrees C and 1.12 +/- 0.3 to 0.52 +/- 0.2 J/g, respectively. The actina values were between 71.0 +/- 0.6 to 75.3 +/- 0.5 degrees C and between 0.5 +/- 0.1 to 0.7 +/- 0.1 J/g. Cooperativity decreased as the storage time increased. This is showing a certain degree of protein displacement. The values found by thermal analyses showed a direct relationship with the functional properties, both decreasing with storage time. PMID- 17425181 TI - [Acceptability and nutritional quality of a beverage based on orange juice and whey powder, preserved by heat or high-intensity pulsed electric fields (HIPEF)]. AB - Orange juice may be a nutrient vehicle that helps to improve diet quality. The addition of whey allows the incorporation of high quality proteins. However, a good acceptability is necessary. The aims of the present study were: a) to assess the acceptability of a beverage prepared with orange juice (J) and whey powder (WP) at 7 g/100 g (J+WP7) or at 13 g/100 g (J+WP13); b) to measure available lysine content and ascorbic acid retention of the more accepted formulation, comparing the effect of HIPEF treatment (29 kV/cm, t(acum): 59 micros) with a conventional heat treatment at 75 degrees C, for 15 minutes (HT). The beverages were subjected to sensory evaluation (Friedman test). Available lysine was assessed by the Carpenter method, modified by Booth, and vitamin C by HPLC; minerals Na and K, by flame photometry; Ca, Mg and Zn by atomic absorption spectrometry. There were no significant differences between the acceptance of J and J+WP7. J+WP13 was significantly less accepted (p < 0.01), so it was discarded. Available lysine (mg/g protein) in untreated J+WP7 was 60.2 +/- 0.15; after treatments: 50.0 +/- 2.8 (HT) and 51.0 +/- 3.4 (HIPEF). The HIPEF treated J and (J+WP7) retained 100% and 98% of their vitamin C initial content and the HT treated, 91% and 88%, respectively. The amount of whey powder added to the orange juice conditioned the acceptability. The juice containing 7% of whey powder was well accepted, and after treatment by HIPEF, it retained a good nutritional quality, regarding available lysine, vitamin C retention and provision of mineral nutrients. PMID- 17425182 TI - [Antioxidant capacity of fruits and vegetables cultivated in Chile]. AB - The high prevalence of non transmissible chronic diseases (NCD) related to food consumption had increased the studies conducted to investigate the relationship between diet and health. A smaller incidence of NCD, with food patterns with high consumption of fruits and vegetables has been observed and chemical compounds of these foods have been one of the main subjects of the actual research in the reaqltion between food consumption and health. The effect of vegetable foods has been attributed to various nutrients and bioactive compounds with antioxidant activity. In order to determine the antioxidant capacity of vegetable foods cultivated in Chile, natural fruits and vegetables were analyzed according to the FRAP (ferric reducing activity power) method, reading to the 4 minutes. In vegetables, the values were between 0.002 and 1.91 milimoles of Fe/l00 g for cooked carrot and red pepper respectively. The values of the fruits ranged between 0.02 milimoles of Fe/100 g for the cucumber and 12.32 for maqui, the berries studies showed values between 3.10 for strawberry and 3.55 for wild blackberry. Lemmon and quince with 0.25 and 0.23 respectively are located in the intermediate level and the lowest values within the fruits corresponded to apple (fuji variety) and peaches. PMID- 17425183 TI - [Brazilian Food Composition Table TBCA-USP (Versions 3 and 4) in the international context]. AB - The Brazilian Food Composition Table (TBCA-USP) aims to divulge information about Brazilian food composition. Therefore, it adopts international patterns that define rules and criteria for the food composition area, through guides and manuals with protocols to be used for data generation and compilation, which amplifies the communication and exchange between laboratories. From 2001 to 2004, 863 new data were introduced in the TBCA-USP, in a total of 1838 data, 1200 of which refers to proximal composition. During this period, the versions 3.0, 4.0 and 4.1 of the TBCA-USP were developed and put on the Internet. These versions had several structural modifications in relation to the ones before. The version 3.0 of the TBCA-USP used to be separated by groups of foods, due to its large number of information. Once this number kept growing, the versions 4.0/4.1 turned to have their food data showed individually, accessed through the name of the food (in Portuguese, English or scientific name). All data is presented as 100 g/food and domestic measurement. There is a form available for compilation of food composition data, in order to stimulate researchers to send and publish data that may be evaluated correctly. The TBCA-USP is being structured similarly to the other international tables that follow patterns of INFOODS (International Network of Food Data Systems); and it is making an effort to create specific databases of nutrients or foods that have functional properties. PMID- 17425184 TI - [Evaluation of the flour and starch from white and purple varieties of mapuey (Dioscorea trifida)]. AB - Mapuey (Dioscorea trifida), is a tropical America tuber, which is appreciated for its taste and fine texture. It has not been fully cultivated in Venezuela, even though products like its flour and starch could replace conventional used products. In this work physical and chemical characteristic of flours from mapuey (varieties white and purple), were assessed, as well as some of their micronutrients. Physical, physicochemical and rheological properties, and chemical composition of isolated starches were also evaluated. Flours were obtained by a drying process and starches by aqueous extraction. Chemical analysis was performed following standard methodologies. The flour yield of purple mapuey was the highest, as was its protein content. The minerals content, showed significant differences between both varieties, presenting purple variety a higher content. Isolated starches showed high purity, this was corroborated by the scanning electron microscopy which showed irregular shaped granules (oval and elongated), with truncated end and smooth surfaces. Purple mapuey granules were smaller. Both varieties exhibited a B type diffraction pattern. The greater swelling power and water absorption capacity was presented by white mapuey, while the highest solubility was shown by the purple one. The white mapuey had maximum viscosity, as well as the highest value of breakdown, suggesting more fragile granules. Setback was lower in the white mapuey, suggesting lower tendency to retrogradation. The purple mapuey although, it showed a higher amylose content, presented lower consistency, even though the difference was not relevant. PMID- 17425185 TI - The future of health care in the United States. PMID- 17425186 TI - Past experiences of the clinical instructor and current attitudes toward evaluation of students. AB - The purpose of this study was to determine whether clinical instructors' past experiences as radiation therapy students impact how they currently evaluate students in the clinical setting. Two survey instruments were mailed to a random sample of 400 credentialed radiation therapists. The first of these questionnaires sought information on past experiences as a student, and the second assessed current attitudes toward clinical evaluation. There were 167 respondents, 85 of who were employed in settings involving the education of student radiation therapists. Among these clinical instructors, statistically significant correlations were found to exist between the two survey instruments that suggest respondents who believe they had positive experiences as radiation therapy students in the clinical setting are more likely to exhibit positive attitudes toward the clinical evaluation of their current students. PMID- 17425187 TI - Factors affecting curriculum content and the integration of evidence-based practice in entry-level physiotherapy programs. AB - Decisions about curricular content in entry-level health professional programs are influenced by a variety of external and internal factors. However, little is known about how lecturers make decisions about the curricular content to be included or excluded from entry-level programs. This study aimed to explore the factors influencing such decision making regarding curricular content in entry level Australian and New Zealand programs for physiotherapy, as well as how evidence-based practice (EBP) is integrated into the teaching and learning framework. Thirteen lecturers from 13 institutions (100% response rate) responsible for teaching a core part of physiotherapy practice, electrophysical agents, participated in a semistructured telephone interview. Decision making for curricular content involved an overall democratic process with the program team, but the day-to-day content was determined by the lecturer. Factors that lecturers reported as impacting on the choice of curriculum were current clinical practice, evidence, and accreditation or registration requirements. Thematic analysis of open-ended questions identified four main themes relating to the integration of the EBP paradigm within teaching: resource materials, use of broad definitions of evidence, inclusion of specific instructional strategies, and context of curriculum. Lecturers used a variety of research methodologies as a backdrop for the presentation of techniques and interventions that are used commonly in clinical practice despite limitations in the evidence base. The results highlighted tensions that exist when designing entry-level curricula with the need to prepare competent and safe practitioners while working within an EBP paradigm. PMID- 17425188 TI - Guiding principles in a merger of allied health and nursing schools. AB - Mergers have long been a reality in higher education during periods of financial challenge. More recently, academic mergers have evolved to become a strategy for achieving academic excellence, broadening institutional vision, and solidifying the competitive position of the merged entities. This report summarizes literature focused on critical considerations when evaluating and implementing mergers in an academic environment using a conceptual model adapted from Kotter. In addition, this paper reports on the planning and initial 9 months of the merger between the Edward and Margaret Doisy School of Allied Health Professions and the School of Nursing to form the Edward and Margaret Doisy College of Health Sciences at Saint Louis University. PMID- 17425189 TI - Eight critical skill sets required for manual therapy competency: a Delphi study and factor analysis of physical therapy educators of manual therapy. AB - Erudition of clinical "skills" used by manually trained physical therapists involves the complex domain of cognitive, affective, and psychomotor concepts. The purpose of the study was to obtain consensus among manual therapy educators regarding stand-alone skills required during competent application of manual therapy and to distill these stand-alone skills into smaller but more homogeneous skill sets. Eighty manual therapy educators in entry-level and post-entry-level educational programs completed a three-round Delphi survey aimed at developing consensus among experts in the field regarding the stand-alone skills that are important to a learners' development of manual therapy competencies. From the Delphi outcomes, a principal component factor analysis distilled eight component skill sets that were associated with orthopedic manual therapy (OMT) proficiency: (1) manual joint assessment, (2) proficiency of fine sensorimotor characteristics, (3) manual patient management, (4) bilateral hand-eye coordination, (5) manual gross characteristics of the upper extremity, (6) manual gross characteristics of the lower extremity, (7) control of self and patient movement, and (8) discriminate touch. Manual joint assessment comprised 46% of the factor analysis variance and contained the majority of stand-alone descriptor statements from a Delphi survey. This study found that despite the disparate backgrounds among respondents, the eight skill sets were identified as important to OMT competency and application. By identifying the key skill sets required for OMT competency and application, OMT educators may more effectively identify teaching methods associated with improved educational outcomes. PMID- 17425190 TI - Gender differences in oral health knowledge and behavior of the health science college students in Kuwait. AB - Dental caries and periodontal diseases have been declining in most industrialized countries, but this positive trend has not been seen in the Middle East. This study aimed to determine oral health knowledge and behavior of the students at the Health Sciences College in Kuwait as well as possible associated factors. This study was first conducted at the college of the male students (n = 153) during the autumn semester in 2001. A similar questionnaire study was then conducted at the college of the female students (n = 547) during the spring semester in 2002. The samples were merged for this study, for a total sample of 700 students. The response rate was 84% (n = 128) among the male students and 73% (n = 400) among the female students. Most of the students had visited a dentist during the past year, and quite a high proportion was seen for an examination or prevention. Female students reported twice-a-day tooth-brushing frequency much more often than did male students. They also used fluoride toothpaste more often than male students. Oral health knowledge (as a summary variable) was statistically significantly higher among the female students than among the male students. It was also strongly associated with the older age among the female students. The knowledge and oral health behavior of the Health Sciences College students in Kuwait, especially among the male students, seems to be poor and calls for an urgent improvement of health education programs. PMID- 17425191 TI - Use of a modified journal club and letters to editors to teach critical appraisal skills. AB - The objective of this study was to report the use of a journal club and letter to the editor writing project to teach critical appraisal skills to health sciences students. Data were collected from 22 consecutive classes (n = 907 students) between 1997 and 2003. The program took place in an 11-week evidence-based practice course in the second year of a chiropractic curriculum. After didactic preparation, students participated in several journal club exercises to appraise papers representing specific research designs. Students then critically appraised a recent article of their choice and wrote a critical appraisal essay in the form of a letter to the editor. The project was evaluated through a review of student assessment, an anonymous survey instrument, and the number of letters submitted and published. Assessments of student performance were consistently high, yielding an average score of 23.7 out of 25 possible points on an objective scale. Responses from 277 students completing a survey on their perceptions of the project were positive, indicating that the goals of the project were being met. Over the 7-year period, approximately 450 letters were written, 12 were submitted for publication, and 10 were published. The results of this study suggest that the use of a journal club and letter-to-the-editor writing project may provide a means by which health care students can gain skills in critical appraisal. Further investigation into these methods should be undertaken. PMID- 17425192 TI - An introduction to the intentional use of hope. AB - Hope, as a health-promoting force, is introduced and distinguished from related concepts to make clear the power of hope. The role of hope in healing and adjustment is explored, and hope-enhancing sources and strategies are described. Suggestions are provided to promote the deliberate use of hope in everyday practice. PMID- 17425193 TI - Protecting consumers in the wake of the U.S. Dietary Supplement Health and Education Act. AB - The Dietary Supplement Health and Education Act of 1994 has given allied health professionals the responsibility of assisting consumers with the use of dietary supplements. Problematic dietary supplements have been marketed and consumed, often for many years, before interventions by the Food and Drug Administration, and the clinical evaluation of these products takes pharmacologic and toxicologic skills that may be beyond the basic competency skills of some allied health professionals. Allied health professionals are in a position to make a stand regarding dietary supplements and their clinical application, but to do so they may need to complement their pharmacologic and toxicologic skills accordingly. PMID- 17425194 TI - How technology is reshaping the practice of nongynecologic cytology: frontiers of cytology symposium. AB - To pay tribute to the Founders of Acta Cytologica, this Golden Anniversary symposium on nongynecologic cytology revives the written symposium style of the 1950s. Participants from countries throughout the world were asked how new technologies are currently applied in their laboratories and whether future advances and challenges can be predicted. The specific questions and the participants' answers follow. PMID- 17425195 TI - Lymph node metastasis of soft tissue tumors: a cytomorphologic study. AB - OBJECTIVE: To study the frequency of regional lymph node metastasis of soft tissue tumors (STT) and to evaluate the utility of fine needle aspiration cytology (FNAC) as an initial investigative modality. STUDY DESIGN: A prospective and retrospective study of over 6 years (1998-2004) was performed to look for frequency of STT metastasizing to lymph nodes. FNAC of enlarged nodes was performed as a routine outpatient procedure after obtaining complete clinical details. Histopathology and immunohistochemistry were correlated where available. RESULTS: Lymph node enlargement was seen in 23 of 241 patients with STTs, of which 19 cases showed involvement (7.88%), synchronous with primary in 12 cases and metachronous in 7 cases. The most common sites of primary tumor were the lower extremity and head and neck region with involved regional lymph nodes. STTs commonly involving lymph nodes were rhabdomyosarcoma and extraskeletal Ewing's/primitive neuroectodermal tumor (PNET); other rare tumors included malignant granular cell tumor, epithelioid hemangioendothelioma, mediastinal ganglioneuroblastoma, angiosarcoma and epithelioid sarcoma. CONCLUSION: Lymph node aspirates should be examined for alien cells, particularly smears that are paucicellular and demonstrate cystic change. Lymph node metastasis of STT is rare and influences staging, treatment and prognosis. Enlarged regional nodes should be examined with FNAC. PMID- 17425196 TI - Frequency and rationale of fine needle aspiration biopsy conversion to core biopsy as a result of onsite evaluation. AB - OBJECTIVE: To measure the frequency and analyze the rationale and potential diagnostic benefits of converting the fine needle aspiration (FNA) procedure to core biopsy. STUDY DESIGN: The frequency of conversion to core biopsy was calculated over 13 months. Analysis of these cases was conducted in regard to the appropriateness for conversion and whether the core biopsy provided additional specific diagnostic information. RESULTS: During this period, the onsite triaging pathologist recommended FNA conversion to core biopsy in 31 of 821 procedures (3.7%). In 3 instances, the core biopsy could not be performed. The rationale for conversion in the remaining 28 cases (3.4%) included either scant aspirated material in 9 cases (32%) or an anticipated need for additional histologic material to further characterize the lesion in the other 19 (68%). In 27 cases (96%), the rationale for conversion was considered to be appropriate, and in 3 of these (11%) the core provided a change in diagnosis. Additional useful diagnostic information was identified in 12 cases (44%). CONCLUSION: Conversion to core biopsy during FNA is infrequent but justified in most cases. Appropriate utilization of this approach is helpful and may be cost effective. PMID- 17425197 TI - The role of fine needle aspiration cytology in evaluation of epididymal nodular lesions. AB - OBJECTIVE: To study the role of fine needle aspiration cytology (FNAC) in evaluation of nodular lesions of the epididymis. STUDY DESIGN: A retrospective study was done on 16 patients who reported with nodular lesions of the epididymis over a 1(1/2)-year period. FNAC was carried out in all cases. Histopathologic correlation was achieved in 10 cases. Cytologic features were studied in all cases and compared with histology results where available. RESULTS: Adequate material was obtained on FNAC in all cases. Cytologic features were adequate to establish a diagnosis in all cases and were confirmed as correct on histologic examination in the 10 cases in which biopsies were performed. Sperm granuloma was the most common lesion encountered. CONCLUSION: FNAC is a rapid, easily available technique for evaluation of nodular lesions of the epididymis. It helps in making a diagnosis and developing a management plan. PMID- 17425198 TI - A cytologic perspective on meibomian gland carcinoma. AB - OBJECTIVE: To reevaluate and reestablish the reproducible diagnostic cytomorphologic features of meibomian gland carcinoma (MGCA), demonstrating the importance of fine needle aspiration cytology (FNAC) in eyelid lesions and leading to better management of the tumor. STUDY DESIGN: Cytologic smears ofpreviously diagnosed cases of MGCA over a 3-year period were collected, along with their histopathology reports. The cytomorphology of the histopathologically proven cases were reevaluated in greater detail. The cytologic smears were stained with Leishman-Giemsa cocktail and Papanicolaou stain, and the histopathologic sections were stained with hematoxylin-eosin. Special stains and immunohistochemical stains were applied as required. RESULTS: Of the 19 cases, 3 diagnosed as MGCA on cytologic examination were reported as basal cell carcinoma on histopathology. On reevaluation of the cytologic smears, in addition to the regular characteristic features of MGCA, interesting findings such as an isolated dispersed cell population in a bubbly background, signet ring cells, pseudo-mucin cell balls, giant cell reaction and hyaline-like round bodies were observed. CONCLUSION: This study emphasizes the cytomorphologic features of MGCA in addition to those described in the literature, which could be of a great help in its diagnosis. Further, it emphasizes the importance of FNAC in the diagnosis of MGCA. PMID- 17425199 TI - Reporting endometrial cells in Papanicolaou smears of women age 40 and older: a qualifying educational note could prevent unnecessary follow-up biopsy. AB - OBJECTIVE: To determine the impact of adding a qualifying educational note when exfoliated endometrial cells were present in the first half of the menstrual cycle. STUDY DESIGN: We identified all Pap smears that had endometrial cells in women > or = 40 years, between July and November, 2004. When endometrial cells were seen in the first half of menstrual cycle, it was stated that, "endometrial cells correlate with the menstrual history provided." We then attempted to determine if this qualifying comment had an impact on the decision to obtain a follow-up endometrial biopsy. RESULTS: A total of 325 women were identified. Of these, 51 (15.7%) had follow-up endometrial biopsy without clinical evidence of endometrial pathology, compared to a rate of 36.9% before implementation of the note (p < 0.0001). Biopsies were performed in 21 of 195 (10.8%) when the note indicated that endometrial cells correlated with menstrual history, compared to 30 of 130 (23%) in whom a different note was appended because endometrial cells were out of phase or patients were postmenopausal (p = 0.0032). Biopsies did not reveal significant endometrial pathology in any of the 51 patients. CONCLUSION: Adding an educational note when endometrial cells are reported in the first half of the menstrual cycle in women > or = 40 years may help reduce unnecessary biopsies. PMID- 17425200 TI - Cytomorphologic characteristics, differential diagnosis and utility during intraoperative consultation for medulloblastoma. AB - OBJECTIVE: To study the cytologic findings that differentiate LC/A and non-LC/A (NLCA) types. STUDY DESIGN: Cytologic smears, prepared during intraoperative consultation (IOC) of 43 cases of pure histologic type, defined as > 80% of the tumor being composed of 1 of 3 major histologic types--classic, nodular and LC/A- were reviewed and graded semiquantitatively (0-3+) for 14 cytologic features: cellularity, nuclear streaming, endothelial proliferation, necrosis, lymphoglandular bodies, rosette formation, apoptosis, nuclear cannibalism, pleomorphism, macronucleoli, paranuclear inclusions, cytoplasmic vacuoles, mitoses and multinucleation. The grades were compared between the 2 histologic groups, LC/A vs. NLCA, as well as between the preparation methods, squash vs. touch (TP). Values of p < 0.05 were accepted as statistically significant. RESULTS: "Cell wrapping" (cannibalism), pleomorphism, macronucleoli, apoptosis, mitoses and cytoplasmic vacuoles were more significantly observed in LC/A than in NLCA. CONCLUSION: Intraoperative cytologic evaluation of these cytologic parameters allows distinction between LC/A and NLCA, which can be prognostically useful during IOC. Rosette formation, the histologic hallmark of classic medulloblastoma, is not a discriminator. Given the fewer artifacts and sufficient cellularity, TP is the preferred method of cytologic preparation. PMID- 17425201 TI - Thin-layer cytology: identification of human papillomavirus by hybridization, capture and signal amplification assay using an alcoholic cell preservative medium. AB - OBJECTIVE: To ensure complete reliability in the detection of hyman papillomavirus using either Specimen Transport Medium (Digene Inc., Gaithersburg, Maryland, U.S.A) or an alcoholic cell preservative. STUDY DESIGN: In order to compare both media, we adjusted the cell content of specimens and optimized the denaturation duration to avoid chemiluminescence inhibition. RESULTS: We validated this protocol in 2 groups of 90 patients, leading to agreement of 100% in both positive and negative tests. This was confirmed using linear regression curves. We demonstrated that any change in the Digene protocol, especially the use of other preservatives, should be carefully evaluated since it may interfere with DNA denaturation and hybridization and the chemiluminescence signal. The main risk was obtaining false positive responses from nonspecific DNA hybridization and false negative ones because of chemiluminescence signal inhibition. CONCLUSION: It is necessary to take into account preservative characteristics when a new protocol is under validation. PMID- 17425202 TI - Diagnosis of pleomorphic adenoma in a heterotopic salivary gland: a case report. AB - BACKGROUND: The histologic diversity encountered in pleomorphic adenoma may cause diagnostic difficulty in fine needle aspiration cytology (FNA) due to limited and selective sampling. CASE: A 40-year-old woman presented with a mass in the anterior aspect of the neck along the sternocleidomastoid muscle. FNA revealed a cellular tumor with a chondrimyxoid background and epithelial cells intermingled with a few mesenchymal cells. The diagnosis of pleomorphic adenoma was confirmed on histopathology. CONCLUSION: Primary ectopic pleomorphic adenoma can confidently be diagnosed by FNA. This technique is a useful tool in the initial assessment of the tumor. PMID- 17425203 TI - Cytologic features of malignant cystic pheochromocytoma: a case report. AB - BACKGROUND: Image-guided fine needle aspiration (FNA) biopsy of deep-seated lesions (e.g., retroperitoneal) has aided preoperative diagnosis and treatment planning. Fine needle aspiration biopsy of pheochromocytomas traditionally has been contraindicated by many authors because of possible hypertensive crisis and fatal hemorrhage. Cytologic features are described for a malignant pheochromocytoma (greater percentage of smaller cells and absence of hyaline globules) with extensive cystic degeneration that was later confirmed by histopathologic, immunohistochemical and ultrastructural studies. CASE: An elderly man was diagnosed with renal cell carcinoma based on clinical examination and ultrasonography. FNA performed on the mass revealed interesting features of pheochromocytoma with an abundance of small cells. CONCLUSION: The cytopathologist should be aware of the cytologic features of pheochromocytoma so as to detect a clinically unsuspected lesion. An increased percentage of smaller cells may be indicative of aggressive tumors. PMID- 17425204 TI - Primary papillary serous carcinoma of the peritoneum: report of a case with diagnosis by fine needle aspiration and immunocytochemistry. AB - BACKGROUND: Primary papillary serous carcinoma (PPSC) of the peritoneum is a rare neoplasm, histologically indistinguishable from papillary serous carcinoma of the ovary, which diffusely involves the peritoneum but spares or minimally invades the ovaries. To the best of our knowledge, the preoperative and the fine needle aspiration diagnosis of this disorder have not been reported before. CASE: A woman developed an extensive peritoneal neoplasm 4 years after hysterectomy and bilateral salpingo-oophorectomy for benign disease. Fine needle aspiration of the tumor was performed, and the cytologic and immunocytochemical findings were consistent with papillary serous carcinoma. A diagnosis of PPSC of the peritoneum was rendered because review of all slides from previous surgical specimens showed no evidence of carcinoma and no other primary tumors were found elsewhere. CONCLUSION: Fine needle aspiration cytology coupled with immunocytochemical and clinical data allows an unequivocal preoperative diagnosis of papillary serous carcinoma (primary peritoneal or with an ovarian origin). The sole limitation to establish a primary peritoneal origin before surgery is the requirement to histologically study the ovaries. Based on this fact, the preoperative fine needle aspiration cytology diagnosis of PSCP should be restricted to oophorectomized patients. PMID- 17425205 TI - Papillary endothelial hyperplasia of the orbit: report of a case highlighting a pitfall on fine needle aspiration biopsy. AB - BACKGROUND: Papillary endothelial hyperplasia (PEH) is an unusual form of thrombus organization that occurs predominantly in the extremities, including the head and neck. However, it is rare in the orbit/ocular region. Although the histologic features of PEH have been well described, the cytologic diagnosis remains difficult. CASE: A 63-year-old man presented with a left intraorbital mass that was increasing in size and associated with paresthesia and a recent history of excision of squamous cell carcinomas (SCC) from his left cheek. Fine needle aspiration biopsy (FNAB) yielded very limited material, predominantly blood. However, 1 Papanicolaou-stained slide showed groups of atypical cells with scanty but dense cytoplasm and large, hyperchromatic nuclei with prominent nucleoli. SCC was favored, and excision was performed. Histology showed PEH within an assumed cystic lymphangioma, associated with inflammatory and fibrotic change in the compressed supraorbital nerve. CONCLUSION: To our knowledge this is the third report on the cytology of PEH and the first report of FNAB in PEH of the orbit. The case was also unusual as it was the second metachronous PEH in the patient. PMID- 17425206 TI - Sebaceous lymphadenoma identified by fine needle aspiration biopsy: a case report. AB - BACKGROUND: Sebaceous lymphadenoma of the parotid gland is a rare benign neoplasm. This is the first reported case of fine needle aspiration biopsy (FNAB) findings for sebaceous lymphadenoma of the parotid gland. CASE: A 60-year-old male presented with painless, bilateral parotid swelling noted for 5 months. The swelling was more pronounced on the right. Examination revealed bilaterally prominent parotid glands with diffuse firmness but no discrete masses. There was no evidence of facial nerve dysfunction. Laboratory evaluation was negative for infectious and autoimmune etiologies. Magnetic resonance imaging revealed bilateral cystic parotid masses. FNAB of the right parotid was obtained to assist with preoperative counseling. It revealed lymphoid and salivary gland parenchymal cells. The patient underwent a right superficial parotidectomy. The surgical specimen of the parotid mass confirmed the diagnosis of sebaceous lymphadenoma on the tissue section. The contralateral parotid mass had not been excised at this writing. CONCLUSION: This report is the first to describe the FNAB findings of the unusual benign parotid neoplasm sebaceous lymphadenoma. Though the definitive diagnosis of any parotid mass requires tissue, generally obtained via parotidectomy, an FNAB diagnosis can be useful in counseling a patient prior to definitive biopsy. PMID- 17425207 TI - Cutaneous metastasis of occult hepatocellular carcinoma: a case report. AB - BACKGROUND: Skin metastases from hepatocellular carcinoma (HCC) represented only 0.8% of all known cutaneous metastases in a recent large series. The most frequent site appears to be the head, but this fact has received little attention. An accurate cytologic diagnosis is extremely difficult in patients with unknown liver dysfunction. We report the cytologic features of a face metastasis from occult HCC. CASE: A 65-year-old woman presented with a mass in the right preauricular region of 2 months' duration. Her past medical history was noncontributory. Fine needle aspiration cytology was performed. Following the cytologic diagnosis, computed tomography revealed a 6-cm mass in the right lobe of the liver, portal vein thrombosis and involvement of the superior vena cava. The smears were very cellular. The most frequent pattern was trabecular with transmural endothelization. The cells had an epithelial appearance and polyhedral shape, exhibiting distinct borders. The nuclei were centrally placed, with a prominent nucleolus. The cytoplasm was granular. There were numerous atypical bare nuclei. Subsequent staining with antihepatocyte showed positivity in most tumor cells. The final diagnosis was metastatic HCC. CONCLUSION: HCCs should be considered in the differential diagnosis of carcinomas metastatic to the face, even in the absence of liver symptoms. PMID- 17425208 TI - Fine needle aspiration of pulmonary adiaspiromycosis: a case report. AB - BACKGROUND: Pulmonary adiaspiromycosis is a common disease of many species of wild rodents and occasionally of humans, caused by the inhalation of spores of the fungus Chrysosporium parvum var crescens (Emmonsia crescens). CASE: A 74-year old female with pulmonary adiaspiromycosis was diagnosed by radiologically guided lung fine needle aspiration (FNA). The specimen showed intracellular and extracellular 100-300 microm conidia with a distinct thick, trilaminar wall, which was positive for Gomori-methenamine silver and periodic acid-Schiff stain. The background consisted of a granulomatous process. CONCLUSION: FNA is an effective method of diagnosing pulmonary adiaspiromycosis, and pathologists need to be aware of the characteristic features of this unusual opportunistic fungal infection. PMID- 17425209 TI - Ectopic lesions as potential pitfalls in fine needle aspiration cytology: a report of 3 cases derived from the thyroid, endometrium and breast. AB - BACKGROUND: Ectopic lesions are rarely encountered. Those that are derived from thyroid, breast, endometrium and salivary glands present with palpable masses that can mimic malignancy. Fine needle aspiration cytology (FNAC) is a practical procedure for the differential diagnosis of such lesions but can reveal surprising images for a cytopathologist. CASES: Three cases of discrete, ectopic lesions at different locations occurred. Case 1 was a 27-year-old woman. Upon diagnosis of a submandibular mass with a diameter of 1 cm, FNAC was performed. The smears showed crowded thyroid follicular cells comprising papillary clusters. A cytologic diagnosis of papillary thyroid lesion was rendered, Histopathology revealed that this lesion was ectopic thyroid tissue with focal chronic thyroiditis. Case 2 was a 38-year-old woman who presented with a painful mass with a diameter of 2.5 cm in the abdominal wall. The patient had undergone cesarean section 3 years earlier. The case was diagnosed on FNAC as low grade malignancy in which an adenocarcinoma/mesenchymal tumor distinction could not be made. The pathologic examination revealed endometriosis. Case 3 was a 31-year old woman who presented with a palpable nodule in the axillary region with a diameter of 1 cm. The patient had given birth 1 month earlier and was nursing. An FNAC diagnosis of lactation ectopic breast tissue was made. The mass disappeared by the end of lactation. CONCLUSION: FNAC of ectopic lesions may prove to be a diagnostic pitfall for cytopathologists. A cytopathologist who encounters a cellularpicturefrom a lesion that is outside the normal anatomic location must use a cautious diagnostic approach. Unless there are clear findings, the cytopathologist must refrain from a diagnosis of malignancy. PMID- 17425210 TI - Spindle cell neuroendocrine carcinoma of the lung: report of a case with fine needle aspiration cytology and differential diagnostic considerations. AB - BACKGROUND: Spindle cell neuroendocrine carcinomas of the lung may be frequently observed on fine needle cytology (FNC) samples and often pose stimulating differential diagnostic problems. CASE: The cytopathologic findings from FNC performed on a long-standing coin lesion of the lung in a 54-year-old woman were analyzed in view of the data and long clinical history. CONCLUSION: A final diagnosis of low grade spindle cell neuroendocrine carcinoma was reached by combining cytopathologic and immunocytochemical information. The main lesions considered in the differential diagnosis were intrapulmonary inflammatory myofibroblastic tumor (fibrohistiocytic variant) and spindle cell thymoma. PMID- 17425211 TI - Myxoinflammatory fibroblastic sarcoma: report of a case with fine needle aspiration cytology. AB - BACKGROUND: Myxoinflammatory fibroblastic sarcoma (MFS) is a distinct neoplasm that usually arises in the acral zones of distalextremities. We report, for the first time, the preoperative fine needle a,spiration cytology (FNAC) findings of an MFS case that was confirmed after surgical excision. CASE: An 81-year-old woman presented with a multinodular tumor in the distal right extremity that had been present for 1 year. FNA C of the lesion was performed and followed by local excision. The fine needle aspiration smears contained 2 of the 3 types of neoplastic cells that have been observed in MFS: spindled and ganglionlike cells. The background was myxoid, with a prominent inflammatory infiltrate. Histopathologic examination of the surgical specimen confirmed the diagnosis of MFS. CONCLUSION: Although the cytologic diagnosis was "pleomorphic sarcoma," MFS was considered and local excision recommended, given the reported low grade nature of this entity. However, the need for extreme caution in the diagnosis of soft tissue lesions on cytologic grounds alone cannot be overemphasized. PMID- 17425212 TI - Use of fine needle aspiration cytology in leprotic lesions: a report of 4 cases. AB - BACKGROUND: Although a few studies have shown fine needle aspiration cytology (FNAC) to be a sensitive diagnostic tool in the detection of nerve involvement, its role as an initial diagnostic procedure in pure neuritic leprosy (PNL) and in the detection of skeletal lesions with unusual findings has not been documented before. CASES: Three patients who presented with thickened nerves and a fourth with biopsy-proven lepromatous leprosy with lesions in hand bones underwent FNAC. Of the 3 patients with nerve thickening, 2 had a clinical suspicion or diagnosis of neuritic leprosy, whereas in the third patient a clinical differential diagnosis of a soft tissue tumor or parasitic cyst was considered. FNAC in all 3 cases revealed epithelioid cell granulomas, Langhans giant cells and caseous necrosis. Fites and Ziehl-Neelsen stains were negative for acid-fast bacilli. Cytologic diagnosis of pure neuritic leprosy was made in all 3 cases and confirmed by histopathologic examination. FNAC of skeletal lesions from the fourth patient confirmed involvement of bone with unusual cytologic findings of epithelioid cell granulomas and giant cells along with a significant proportion of foamy macrophages and strong Fites stain positivity. CONCLUSION: FNAC is a simple, useful, minimally traumatic and routinely applicable procedure in the diagnosis of pure neuritic leprosy and leprous osteitis. PMID- 17425213 TI - Fine needle aspiration biopsy and immunostaining findings in an aggressive inflammatory myofibroblastic tumor of the lung: a case report. AB - BACKGROUND: Inflammatory myofibroblastic tumors (IMTs) can vary from benign pseudosarcomatous tumors to low grade sarcomas. To date, fine needle aspiration (FNA) findings of lung IMTs, especially in the aggressive form, have not been fully described. Here we present FNA biopsy findings in conjunction with immunohistochemical studies in a case of primary and recurrent pulmonary IMT. CASE: A 22-year-old man first presented with a left lung mass and 4.5 years later with a recurrent mass. Preoperative computed tomography-guided FNA was performed on both tumors. FNA cytologic smears of both specimens consisted of scant, distorted spindle cells suggestive of a spindle cell lesion but were insufficient for further classification. Needle core biopsies as well as touch imprints were performed during the FNA procedures. The imprints revealed abundant, well preserved spindle cells with mild to moderate atypia and intermixed lymphocytes and plasma cells. The spindle cells in both specimens were immunoreactive for vimentin and smooth muscle actin and were negative for pancytokeratin, desmin, CD34 and c-kit. Thirty percent of the tumor cells were positive for p53. The findings were compatible with those of IMT. Histologic examination of the surgically resected initial and recurrent masses confirmed the diagnosis of lMT. CONCLUSION: The cytologic findings of pulmonary IMT in FNA specimens are suggestive of, although not specific for, IMT. Immunohistochemical studies can assist in the diagnosis by excluding other spindle cell lesions. Cytologic atypia and p53 immunoreactivity may be indicators of aggressive IMTs. PMID- 17425214 TI - Erythrocytes as funguslike artifacts in pulmonary cytology. PMID- 17425215 TI - Filariasis coexisting with a cold abscess. PMID- 17425216 TI - Cholesterol crystals in the thyroid gland. PMID- 17425217 TI - Hematopoietic cells in thyroid fine needle aspirates. PMID- 17425218 TI - Verrucous papillomatous lesion of post-kala-azar dermal leishmaniasis: fine needle aspiration diagnosis of an unusual presentation. PMID- 17425219 TI - Academic sleep centers of the future: an achievable vision. PMID- 17425220 TI - Altered circadian and homeostatic sleep regulation in prokineticin 2-deficient mice. AB - STUDY OBJECTIVES: Sleep is regulated by circadian and homeostatic processes. Recent studies with mutant mice have indicated that circadian-related genes regulate sleep amount, as well as the timing of sleep. Thus a direct link between circadian and homeostatic regulation of sleep may exist, at least at the molecular level. Prokineticin 2 (PK2), which oscillates daily with high amplitude in the suprachiasmatic nuclei (SCN), has been postulated to be an SCN output molecule. In particular, mice lacking the PK2 gene (PK2-/-) have been shown to display significantly reduced rhythmicity for a variety of circadian physiological and behavioral parameters. We investigated the role of PK2 in sleep regulation. DESIGN: EEG/EMG sleep-wake patterns were recorded in PK2-/- mice and their wild-type littermate controls under baseline and challenged conditions. MEASUREMENTS AND RESULTS: PK2-/- mice exhibited reduced total sleep time under entrained light-dark and constant darkness conditions. The reduced sleep time in PK2-/- mice occurred predominantly during the light period and was entirely due to a decrease in non-rapid eye movement (NREM) sleep time. However, PK2-/- mice showed increased rapid eye movement (REM) sleep time in both light and dark periods. After sleep deprivation, compensatory rebound in NREM sleep, REM sleep, and EEG delta power was attenuated in PK2-/- mice. In addition, PK2-/- mice had an impaired response to sleep disturbance caused by cage change in the light phase. CONCLUSIONS: These results indicate that PK2 plays roles in both circadian and homeostatic regulation of sleep. PK2 may also be involved in maintaining the awake state in the presence of behavioral challenges. PMID- 17425221 TI - Long term effects of sleep deprivation on the mammalian circadian pacemaker. AB - STUDY OBJECTIVES: In mammals, sleep is controlled by a homeostatic process, which regulates depth of sleep, and by the circadian clock of the suprachiasmatic nucleus (SCN), which regulates 24-h rhythms in timing of sleep. Sleep deprivation is known to cause molecular and physiological changes and results in an alteration in the timing of sleep. It is generally assumed that following sleep deprivation, homeostatic mechanisms overrule the circadian clock, allowing animals to sleep during their active phase. However, recent evidence indicates that sleep states have direct access to the circadian pacemaker of the SCN. We questioned therefore whether sleep deprivation may have long-term effects on the circadian pacemaker, which may explain altered sleep patterns following sleep deprivation. DESIGN: To test this hypothesis, we combined SCN recordings of electrical impulse frequency through stationary implanted electrodes in freely moving rats with electroencephalogram recordings in the same animal before, during, and after a mild 6-h sleep deprivation. MEASUREMENTS AND RESULTS: Following sleep deprivation, SCN neuronal activity was significantly reduced to about 60% of baseline levels. The decrements in SCN activity were most obvious during NREM sleep and REM sleep and lasted for 6-7 hours. CONCLUSIONS: The data show that sleep deprivation influences not only sleep homeostatic mechanisms, but also SCN electrical activity, resulting in a strong reduction in circadian amplitude in the major output signal from the SCN. PMID- 17425222 TI - The direct and indirect costs of untreated insomnia in adults in the United States. AB - OBJECTIVES: To estimate the direct and indirect cost burden of untreated insomnia among younger adults (age 18-64), and to estimate the direct costs of untreated insomnia for elderly patients (age 65 and over). DESIGN: A retrospective, observational study comparing insomnia patients to matched samples without insomnia. SETTINGS: Self-insured, employer sponsored health insurance plans in the U.S. PATIENTS OR PARTICIPANTS: 138,820 younger adults and 75,558 elderly patients with insomnia, plus equal-sized, matched comparison groups. INTERVENTIONS: NA. MEASUREMENTS AND RESULTS: Direct costs included inpatient, outpatient, pharmacy, and emergency room costs for all diseases, for six months before an index date. The index date for insomnia patients was the date of diagnosis with or the onset of prescription treatment for insomnia, some-time during July 1, 1999-June 30, 2003. Non-insomnia patients were assigned the same index dates as the insomnia patients to whom they were matched. Indirect costs included costs related to absenteeism from work and the use of short-term disability programs. Propensity score matching was used to find insomnia and non insomnia patients who had similar demographics, location, health plan type, comorbidities, and drug use patterns. Regression analyses controlled for factors that were different even after matching was completed. We found that average direct and indirect costs for younger adults with insomnia were about $1,253 greater than for patients without insomnia. Among the elderly, direct costs were about $1,143 greater for insomnia patients. CONCLUSIONS: Insomnia is associated with a significant economic burden for younger and older patients. PMID- 17425223 TI - Epidemiology of insomnia: a longitudinal study in a UK population. AB - STUDY OBJECTIVES: To investigate the incidence, persistence, and consequences of insomnia and their associations with psychological health and pain. DESIGN: A population based, longitudinal, cohort study using postal questionnaires at baseline and 12-month follow-up. Sleep problems in the past month were assessed using 4 questions: insomnia was defined as having at least 1 of the sleep problems "on most nights." Questions about psychological health, presence of pain at different sites, and demographic details were included in the questionnaire. SETTING: Five general practices in Staffordshire, UK. PARTICIPANTS: The questionnaire was mailed to a random sample of 4885 adults aged 18 years and over registered with these practices. There were 2662 questionnaires returned. RESULTS: Of the responders, 2363 completed all 4 sleep questions at baseline: 870 (37%) had insomnia and 1493 (63%) did not have insomnia. Of those without insomnia at baseline, the incidence of insomnia at 12 months was 15%, and this was significantly associated with baseline anxiety, depression, and pain. Of those who did have insomnia at baseline, 69% had insomnia at 12-month follow-up; persistence of insomnia was significantly associated with older age. Insomnia at baseline was significantly associated with incidence of anxiety, depression, and widespread pain at 12-month follow-up. CONCLUSIONS: Insomnia is common and often persistent. Older people appear more vulnerable to persistent symptoms. Our results provide evidence that the common problems of insomnia, pain, and psychological distress are intertwined and suggest that combined approaches to treatment may be needed to reduce the onset and persistence of these problems in the community. PMID- 17425224 TI - Effect of short-term treatment with gaboxadol on sleep maintenance and initiation in patients with primary insomnia. AB - STUDY OBJECTIVE: To perform an early evaluation of the efficacy and safety of gaboxadol in the treatment of primary insomnia. METHODS: 26 adults (18-65 years) with DSM-IV criteria for primary insomnia were randomly assigned gaboxadol (5 mg, 15 mg) or placebo in a double-blind, crossover study. After a 3-night polysomnographic (PSG) screen, treatment was administered 30 min before bedtime on 2 consecutive nights during 3 separate sessions including PSG. Efficacy analyses (n = 23) were based on the average of Nights 1 and 2, and compared gaboxadol versus placebo. Baseline was the average of Nights 2 and 3 of the screening session. Both gaboxadol doses significantly (P < 0.05) improved mean total sleep time (mean +/- SD: baseline = 368.0 +/- 51.1 min, 15 mg = 420.3 +/- 24.5 min, 5 mg = 419.8 +/- 20.4 min, placebo = 408.7 +/- 30.4 min). Both gaboxadol doses reduced mean wake after sleep onset, although statistical significance was only achieved with 5 mg (baseline = 61.6 +/- 35.4 min, 15 mg = 38.0 +/- 21.1 min, 5 mg = 34.6 +/- 14.3 min, placebo = 43.4 +/- 22.9 min). Gaboxadol 15 mg also significantly reduced mean latency to persistent sleep (baseline = 55.6 +/- 27.0 min, 15 mg = 23.6 +/- 15.1 min, placebo = 30.0 +/- 19.1 min) and enhanced slow wave duration (baseline = 72.4 +/- 20.8 min, 15 mg = 114.0 +/- 37.5 min, placebo = 93.9 +/- 31.3 min) with no significant effects on REM sleep duration. Patient reports (Leeds Sleep Evaluation Questionnaire) of reduced time to sleep and increased sleep quality showed significant improvement with gaboxadol 15 mg. No next-day residual effects were observed with either dose of gaboxadol (assessed 2 h and 9 h after lights on). All adverse events were mild or moderate. CONCLUSION: Gaboxadol 15 mg was effective and generally well tolerated in the short-term treatment of patients with primary insomnia. Gaboxadol also enhanced slow wave sleep duration and had no significant effects on REM sleep duration. These findings suggest that gaboxadol may be a useful treatment for insomnia. PMID- 17425225 TI - Sleep-disordered breathing and cardiovascular risk. AB - Sleep-disordered breathing, broadly characterized by obstructive sleep apnea (OSA) and central sleep apnea (CSA), is an increasingly recognized public health burden. OSA, consisting of apneas or hypopneas associated with respiratory efforts in the face of upper airway narrowing or collapse, is a common disorder that can be effectively treated with continuous positive airway pressure (CPAP). OSA not only results in daytime sleepiness and impaired executive function, but also has been implicated as a possible cause of systemic disease, particularly of the cardiovascular system. CSA, which may coexist with OSA, has gained attention because of the association of Cheyne-Stokes respiration with an ever-increasing prevalence of heart failure in an aging population. This article reviews some of the extensive literature on pathophysiologic mechanisms as they may relate to the development of cardiac and vascular disease and examine the evidence suggesting OSA as a specific cause of certain cardiovascular conditions. Available evidence regarding the implications of CSA in the context of heart failure is discussed. PMID- 17425226 TI - Proton magnetic resonance spectroscopy study of brain metabolism in obstructive sleep apnoea syndrome before and after continuous positive airway pressure treatment. AB - STUDY OBJECTIVES: Obstructive sleep apnoea syndrome (OSAS) causes sleep related oxygen desaturation, excessive daytime sleepiness (EDS), and cognitive impairment. The role of hypoxic brain damage, sleep fragmentation, and the associated comorbidities (hypertension, vascular disorders) in the pathogenesis of cognitive deficits remains controversial. The aim of this study was to evaluate the cerebral metabolism of OSAS patients in vivo before and after CPAP treatment. DESIGN AND PATIENTS: Fourteen OSAS patients without cardiovascular or cerebrovascular impairment underwent the same protocol before and after 6 months of CPAP including: overnight videopolysomnography (VPSG), Multiple Sleep Latency Test (MSLT), and within the next 2 days neuropsychological and 1H-MRS evaluations. Single voxel 1H-MRS was performed in the parietal-occipital cortex, and absolute concentrations of N-acetyl-aspartate (NAA), creatine, and choline were measured, acquiring spectra at multiple echo-times and using water as internal standard. Ten matched controls were also studied. RESULTS: OSAS patients had a mean RDI of 58/hr, a mean arousal index of 57/hr, and a mean nadir SpO2 of 71%. Before CPAP, all patients showed a normal global cognitive functioning, with only a small number of pathological tasks in working memory and attention tests in a minority of patients. CPAP therapy was effective in resolving sleep apnoea and normalizing sleep structure, and improving EDS and neuropsychological alterations. Before CPAP treatment cortical [NAA] in OSAS (11.86 mM +/- 0.80, mean +/- SD) was significantly lower than in controls (12.85 +/- 0.93; P = 0.01) and positively correlated with minimum SpO2 during sleep (r = 0.69; P = 0.006) and MSLT scores (r = 0.62; P = 0.01). Cortical [NAA] reduction persisted after therapy (11.94 +/- 1.33; P = 0.87 versus pre-CPAP). CONCLUSIONS: OSAS patients have cortical metabolic changes consistent with neuronal loss even in the absence of vascular comorbidities. Metabolic changes persisted after CPAP in the absence of EDS, nocturnal arousals, and major cognitive deficits, likely related to hypoxic damage prior to CPAP treatment. PMID- 17425227 TI - Gender-related differences in symptoms of patients with suspected breathing disorders in sleep: a clinical population study using the sleep disorders questionnaire. AB - BACKGROUND: Gender-related differences in the symptom profile of patients with suspected sleep disordered breathing (SDB) may be one explanation of the clinical underrecognition of SDB in women. STUDY OBJECTIVES: The aim of this study was to prospectively assess gender-related differences in presenting symptoms in a clinical sample of patients with suspected sleep disordered breathing. DESIGN: Administration of the Sleep Disorders Questionnaire prior to clinical and polysomnographic evaluation. Responses obtained from the questionnaire were used to construct 4 independent symptom scales: sleep apnea (SA), periodic limb movement syndrome (PLM), psychiatric sleep disorder (PSY), and narcolepsy (NAR). Analyses of variance were used to examine the effect of gender, AHI, and age on the symptom scales. Associations between gender and each diagnostic scale of the questionnaire were determined by multiple analyses of covariance. SETTING: Tertiary pulmonary referral center. PARTICIPANTS: 2739 men and 782 women with suspected SDB. All patients who were referred to the sleep laboratory underwent full-night polysomnography, irrespective of the likelihood of SDB. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Overall, men scored significantly higher on items related to worsening of snoring/breathing with alcohol (P < 0.001) and smoking history (P < 0.01) than women. Complaints such as witnessed apneas (P < 0.001) and worsening of snoring in supine position (P < 0.05), however, were more frequently reported by men with an apnea-hypopnea index (AHI) < 5/hr, compared with AHI-matched women. There were no significant differences in these items in patients with an AHI > 15/hr. In contrast, women complained significantly more often of insomnia, restless legs, depression, nightmares, palpitations at night, and hallucinations than men. As a result, women had significantly higher scores on the PLM, PSY, and NAR scales of the Sleep Disorders Questionnaire (P < 0.001, for all). After adjustments for age, body mass index, AHI, arousal index, oxygen saturation data, and smoking history, by means of multiple analyses of covariances, gender differences remained significant (P < 0.001, for all scales). CONCLUSIONS: We observed significant gender-related differences in presenting symptoms of patients with sleep disordered breathing at a tertiary level. These differences should be taken into consideration in clinical evaluation of women with suspected sleep disordered breathing. PMID- 17425228 TI - Early CPAP use identifies subsequent adherence to CPAP therapy. AB - STUDY OBJECTIVES: To explore the relationship between specific factors such as sex and early continuous positive airway pressure (CPAP) use, and 30-day adherence to CPAP therapy. DESIGN AND SETTING: Retrospective study conducted at a single center in southeast Michigan. PATIENTS: One hundred patients with obstructive sleep apnea who were recently initiated on CPAP therapy with electronic adherence information relayed from the CPAP device to a laboratory based computer through telephone modem. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: An empiric threshold value of objective CPAP use of greater than 4 hours per night measured 3 days following CPAP initiation was predictive of level of CPAP adherence measured 30 days later. Furthermore, CPAP adherence was directly proportional to age (R = 0.25, P = .018). There were no sex-related differences in adherence to CPAP therapy. CONCLUSIONS: Long-term adherence to CPAP therapy can be predicted as early as 3 days following CPAP initiation. The study also demonstrates that younger age and African-American race are independently associated with lower CPAP adherence. PMID- 17425229 TI - Maintenance of wakefulness test as a predictor of driving performance in patients with untreated obstructive sleep apnea. AB - STUDY OBJECTIVE: To determine the ability of Maintenance of Wakefulness Test (MWT) to predict simulated driving performance in patients suffering from sleep apnea syndrome. DESIGN: Study involving one hour of simulated driving, one night of polysomnography (PSG), and a 4 x 40-minute MWT. SETTING: Sleep laboratory. PATIENTS: Thirty male patients with untreated obstructive sleep apnea syndrome (OSAS) (mean age [+/- SD] = 51 +/- 8 years, range 34-62; mean body mass index (BMI) [+/- SD] = 29 +/- 3, range 24-37; mean apnea/hypopnea index (AHI) [+/- SD] = 43 +/- 24, range 14-96). As defined by MWT mean sleep latency, 23.3% of the patients were sleepy (0-19 min), 33.3% were alert (20-33 min), and 43.4% were fully alert (34-40 min). MEASUREMENTS: Nocturnal PSG, mean sleep latency at 4 x 40-minute MWT trials, Epworth Sleepiness Scale (ESS), and standard deviation from the center of the road (SDS) on driving simulator. RESULTS: Mean MWT scores inversely correlated with SDS during the simulated driving session (Pearson's r = -0.513, P < 0.01). We found a significant effect of MWT groups (sleepy, alert, or fully alert) on SDS (ANOVA, F(2, 29) = 5.861, P < 0.01). Post hoc tests revealed that the sleepy group had a higher SDS than the fully alert group (P = 0.006). ESS, AHI, microarousal index, and total sleep time did not predict simulated driving performance. CONCLUSIONS: A pathological MWT mean sleep latency (0-19 min) is associated with simulated driving impairment. Before MWT can be used to predict the driving ability of untreated patients with OSAS, further studies are needed to confirm that pathological MWT scores are associated with real driving impairment. PMID- 17425230 TI - Sleepy driver near-misses may predict accident risks. AB - STUDY OBJECTIVES: To quantify the prevalence of self-reported near-miss sleepy driving accidents and their association with self-reported actual driving accidents. DESIGN: A prospective cross-sectional internet-linked survey on driving behaviors. SETTING: Dateline NBC News website. RESULTS: Results are given on 35,217 (88% of sample) individuals with a mean age of 37.2 +/- 13 years, 54.8% women, and 87% white. The risk of at least one accident increased monotonically from 23.2% if there were no near-miss sleepy accidents to 44.5% if there were > or = 4 near-miss sleepy accidents (P < 0.0001). After covariate adjustments, subjects who reported at least one near-miss sleepy accident were 1.13 (95% CI, 1.10 to 1.16) times as likely to have reported at least one actual accident as subjects reporting no near-miss sleepy accidents (P < 0.0001). The odds of reporting at least one actual accident in those reporting > or = 4 near-miss sleepy accidents as compared to those reporting no near-miss sleepy accidents was 1.87 (95% CI, 1.64 to 2.14). Furthermore, after adjustments, the summary Epworth Sleepiness Scale (ESS) score had an independent association with having a near miss or actual accident. An increase of 1 unit of ESS was associated with a covariate adjusted 4.4% increase of having at least one accident (P < 0.0001). CONCLUSION: A statistically significant dose-response was seen between the numbers of self-reported sleepy near-miss accidents and an actual accident. These findings suggest that sleepy near-misses may be dangerous precursors to an actual accident. PMID- 17425231 TI - The effects of 53 hours of sleep deprivation on moral judgment. AB - STUDY OBJECTIVES: Functional neuroimaging studies suggest a prominent role for the medial prefrontal cortex in the formation of moral judgments. Activity in this region has also been shown to decline significantly during sleep loss. We therefore examined the effects of 2 nights of sleep deprivation on several aspects of moral judgment. DESIGN: Participants made judgments about the "appropriateness" of various courses of action in response to 3 types of moral dilemmas at rested baseline and again following 53 hours of continuous wakefulness. SETTING: In-residence sleep laboratory at the Walter Reed Army Institute of Research. PARTICIPANTS: Twenty-six healthy adults (21 men, 5 women). INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Compared to baseline, sleep deprivation resulted in significantly longer response latencies (suggesting greater difficulty deciding upon a course of action) only for Moral Personal (i.e., emotionally evocative) dilemmas, whereas response times to Moral Impersonal (less emotionally evocative) and Non Moral dilemmas did not change significantly with sleep loss. The effect of sleep deprivation on the willingness to agree with solutions that violate personally held moral beliefs was moderated by the level of emotional intelligence, as measured by the Bar-On EQ-i. Persons high in emotional intelligence were less susceptible to changes in moral judgments as a function of sleep loss. CONCLUSIONS: These findings suggest that sleep deprivation impairs the ability to integrate emotion and cognition to guide moral judgments, although susceptibility to the effects of sleep loss on this ability is moderated by the level of emotional intelligence. PMID- 17425232 TI - The characteristics of recovery sleep when recovery opportunity is restricted. AB - STUDY OBJECTIVES: The aim of this study was to investigate the recovery of sleep and waking functions following one night of total sleep deprivation, when recovery opportunity was either augmented or restricted. DESIGN: The 9-day laboratory study involved a baseline night, a night of sleep loss (approximately 40 h) followed by 5 nighttime recovery sleep periods. Recovery consisted of either five 9-h sleep opportunities or five 6-h sleep opportunities. SETTING: All data collection took place in a controlled laboratory environment at the Centre for Sleep Research. PARTICIPANTS: A total of n = 20 healthy adults (18-35 yrs) participated in the study. RESULTS: Each sleep period was recorded using a standard polysomnographic EEG montage. Waking functions were assessed every 2 hours during all wake periods, using a 10 minute psychomotor vigilance task (PVT) and a subjective alertness visual analogue scale (VAS). Sleep analyses indicated that across the week TST, SOL, REM, and sleep efficiency varied significantly between the 2 conditions, but amounts of SWS did not. Waking functions in the 9-h condition recovered after one sleep period. In the 6-h condition however, mean response time on the PVT was 10% below baseline and subjective alertness 20% below baseline for the entire recovery period. CONCLUSIONS: The results suggest that TST is a key factor in determining recovery. When recovery opportunity is restricted, both sleep variables and measures of waking functions do not recover. PMID- 17425233 TI - The impact of media use on sleep patterns and sleep disorders among school-aged children in China. AB - STUDY OBJECTIVES: To determine the amount of television viewing and computer use in urban school-aged Chinese children, and to examine their associations with sleep/wake patterns, duration of sleep, and sleep disorders. DESIGN AND SETTING: Students representing 8 Chinese cities were studied during November and December 2005. PARTICIPANTS: A total of 19,299 elementary-school children, 49.7% boys and 50.3% girls, with a mean age of 9.00 years, participated in the survey. MEASUREMENTS: A parent-administered questionnaire and the Chinese version of the Children's Sleep Habits Questionnaire were completed to quantify media use and to characterize sleep patterns and sleep disturbances. RESULTS: A television or computer was present in the bedroom of 18.5% and 18.3% of Chinese school-aged children, respectively. Media presence in the bedroom and media use were positively correlated with later bedtimes, later awakening times, and a shorter duration of sleep during weekdays and weekends. They were also significantly associated with at least 2 types of sleep disturbances. Overall, the most affected sleep behaviors were bedtime and awakening time on the weekends, the duration of sleep during the weekdays, and sleep disorders of bedtime resistance and sleep anxiety. Television viewing > or = 2 hours/day on weekends, with a prevalence of 48.8%, was the predominant risk factor for all sleep disorders with the exception of the sleep duration disorder. Computer use, however, had no correlation with any sleep disorder. CONCLUSION: The presence of media in a child's bedroom and media use had a negative effect on children's sleep/wake patterns, duration of sleep, and sleep disorders. PMID- 17425234 TI - Improving access to specialty care. AB - BACKGROUND: Waits and delays plague health care systems worldwide, and wait times for most specialists exceed those for primary care practices. In office-based practices, the provider office presence is not diluted by competing indispensable activities, and the demand for service is most often for a single type, or stream, of office-based appointment demand. In the more complex specialty practices, however, the demand streams for office visits and other services compete for provider time and dilute the supply of office visits. SEVEN FLOW STRATEGIES, WITH A FOCUS ON THE INITIAL APPOINTMENT: Seven strategies for reduction of delay can be applied, not only at all steps in patient flow and for all demand streams but also at all steps (for example, office visit, diagnostic procedure, surgery, follow-up) and within all specialty care types and ranges of practice. Each specialty care practice will need to discover how to use the basic principles and implement customized solutions within its own unique environment. Although it is ultimately critical to eliminate the delays in all streams of service, the focus is on the application of change strategies at the initial step between primary care and all specialty care practice types. The strategies are (1) balance supply and demand at each step in the chain, (2) work down the backlog, (3) reduce appointment types, (4) independent contingency planning for all variation, (5) reduce the demand for visits, (6) increase the supply, and (7) improve the efficiency of the office work flow. SUMMARY: Specialists support various, distinct demand streams that require demand/supply balance to achieve optimal system performance. If demand/supply balance exists within any stream, waits can be minimized, and the practice can choose time frames within which to balance workload. PMID- 17425235 TI - Do "most wired" hospitals deliver better care? AB - BACKGROUND: A study was conducted in 2006 to compare differences in objective quality of care measures among hospitals labeled "Most Wired"--a hospital or member-hospital of a health system listed among the Hospital and Health Network's Healthcare's Most Wired Hospitals for 2004--versus hospitals without that designation. METHODS: Ten quality indicators representing cardiac and pulmonary measures were calculated for adult hospitals participating in the U.S. Department of Health and Human Services' Hospital Compare initiative. Performance of Most Wired hospitals and comparison hospitals was compared using t-tests. The association of the Most Wired designation to measures of care was assessed using multivariable linear regression and generalized estimating equations. RESULTS: Compared with comparison hospitals, Most Wired hospitals tend to be larger, not for profit and teaching hospitals. Most Wired hospitals outperformed comparison hospitals in all but one quality indicator (p < .05). After adjustment, Most Wired hospitals were independently associated with better quality scores for only 2 out of 10 quality indicators. The Most Wired hospitals did not significantly underperform for any indicator. CONCLUSION: Most Wired hospitals outperformed other hospitals on most objective quality of care measures. However, some of the results were significantly attenuated by other factors associated with quality, suggesting that for specific indicators, "Most Wired" may be a marker of overall quality more than an independent factor. More research is needed on how overall implementation of health information technology directly affects quality of care measures. PMID- 17425236 TI - Eliminating preventable death at Ascension Health. AB - BACKGROUND: Borgess Medical Center, the alpha site, developed innovative strategies to eliminate preventable deaths--one of Ascension Health's eight priorities for action. IMPLEMENTATION OF STRATEGIES: A multifocal approach included the intensivist and hospitalist models and six strategies, four in critical care and two outside critical care. RESULTS: The results of one critical care strategy--tight glycemic control with insulin drips--and one non-critical care strategy--deploying rapid response teams--are reported for three periods: Period 1 (baseline; April 1, 2003-March 31, 2004), Period 2 (April 1, 2004-March 31, 2005), and Period 3 (April 1, 2005-March 31, 2006). Hyperglycemia (> 150 mg/dL) decreased from 48.12% to 37.18% (Period 2) and 25.08% (Period 3). Hypoglycemia (< 70 mg/dL), beginning at 1.64%, increased moderately, to 1.69% and 2.15%. Rapid response team calls per 1,000 discharges increased by 77.54% from Period 2 to Period 3--from 6.28 to 11.15. With an overall 25% reduction in mortality rate as a realistic expression of "eliminating preventable death" by 2008, observed mortality decreased during a two-year period by 19.2% (+/- 0.74%). DISCUSSION: Decreases in mortality were accompanied by control of hyperglycemia in critical care and the implementation of rapid response teams to rescue patients before cardiopulmonary arrest outside of critical care. Most of the preventable deaths occurred among non-end-of-life-care patients and were associated with adverse events, which in most cases, had been identified within Ascension Health's priorities for action. PMID- 17425237 TI - New technology, new errors: how to prime an upgrade of an insulin infusion pump. AB - BACKGROUND: A series of use errors occurred when switching an ambulatory care clinic patient from an older model to a newer model of an ambulatory continuous subcutaneous insulin infusion pump. CASE REPORT: The nurse practitioner (N.P.) reviewed the new pump's mechanics with the patient, who had a 26-year history of Type 1 diabetes mellitus, and supervised the patient's programming of the pump. At bedtime, a blood sugar of > 250 mg/dL prompted the patient to give herself insulin via the pump. The next morning, she was treated at the emergency department for diabetic ketoacidosis. CASE ANALYSIS: The pump had been improperly primed, resulting in no insulin delivery. The incident also reflected the absence of a fail-safe mechanism(s) on the pump to alert the user to the improper priming and inappropriate handoff of the patient's care. Unlike the old pump, the new pump did not require manual priming. The lack of delivery of insulin resulted in DKA, a potentially life-threatening complication of diabetes. A root cause analysis suggested several important safety issues, including skipping of steps on the patient training checklist and other shortcuts in patient training. DISCUSSION: The clinic developed policies and procedures, including mandatory formal training for each pump model by the certified pump trainer and for initiation of insulin pump therapy. This case illustrates the importance of a structured device selection process, provider education, patient education, and monitoring for safety and effectiveness of technological devices in care. PMID- 17425238 TI - The business case for quality at a university teaching hospital. PMID- 17425239 TI - How "user friendly" is the hospital for practicing hand hygiene? An ergonomic evaluation. AB - BACKGROUND: An important cause of nonoptimal hand hygiene may be lack of "user friendliness" of hand hygiene resources due to violation of ergonomic principles in the design of the hospital environment and lack of timely replenishment of consumable resources. METHODS: An ergonomics-based tool, SWAG (for the four main hand hygiene resources-sinks, waste receptacles, alcohol-based hand rub dispensers, and gloves) was developed and implemented to assess the intensive care units and 59 individual rooms in the hospital for structural ergonomic characteristics that facilitate usage of these resources. RESULTS: Several deficiencies in the structural layout of hand hygiene resources were identified that hinder their usage, such as poor visibility, difficulty of access, placement at undesirable height, lack of redundancy, and wide spatial separation of resources that are used sequentially. Consumable hand hygiene resources were often not available because of lack of timely replenishment. DISCUSSION: Many simple inexpensive changes using ergonomic principles can be implemented to promote hand hygiene in hospitals. PMID- 17425240 TI - [OralCDx brush biopsy--a tool for early diagnosis of oral squamous cell carcinoma]. AB - Aim of the present study was an evaluation of the OralCDx brush biopsy (CDx Laboratories Germany, D-72764 Reutlingen) as a screening method for apparently benign lesions of the oral mucosa which due to their harmless clinical appearance previously would not have been subjected to incisional biopsy. One hundred and eighteen brush biopsies from 100 patients were analyzed. Ninety-three brush biopsies from 79 patients revealed a negative result. All OralCDx "atypical", "positive" and "inadequate" results were referred for conventional scalpel biopsy and examined by conventional histology. In six patients (seven OralCDx tests) with the result "positive" and in 14 patients with the result "atypical" by histological examination, carcinoma (4), severe dysplasia (4), moderate dysplasia (1) and mild dysplasia (4) were detected. The OralCDx brush biopsy proved to be a valuable new minimally invasive method for early detection and surveillance of oral dysplasia and of oral squamous cell carcinoma of innocuous appearance. PMID- 17425241 TI - [Multiple cemento-ossifying fibromas of the jaw: a very rare diagnosis]. AB - The cemento-ossifying fibromas (COF) of the jaws are well circumscribed, generally slow-growing, benign lesions which enlarge in an expansive manner. On occasion, they may reach a large size and may result in considerable deformity. The histological pattern of these lesions varies with the stages. In most reported cases ossifying and cemento-ossifying fibromas occur as a solitary lesion. Multiple occurrence of such lesions is rare. The term"cemento-ossifying fibroma"is used to describe fibrous lesions containing calcifications with strong similarity between bone and cementum. Although WHO and some authors regard the cementifying fibroma (CF) as an odontogenic tumor and consider ossifying fibroma (OF) separately as non-odontogenic neoplasm, there is general agreement that CF and OF represent only histologic variants of the same lesion. The case of a 36 year old woman with multiple cemento-ossifying fibromas of the mandible and maxilla demonstrates the diagnostic procedures and a possible therapeutic strategy for this rare lesion. PMID- 17425242 TI - [Tobacco use prevention and cessation in the dental practice]. AB - This is the fourth part of a series of publications from the Swiss task force named "Smoking--intervention in the private dental office" on the topic "tobacco use and dental medicine". It presents the implementation of tobacco use prevention and cessation in the dental practice. Next to the optimal performance of plaque control, tobacco use cessation has become the most important measure for the treatment of periodontal diseases. In contrast to general medicine practice, the dental practice team is seeing its patients regularly and is therefore capable of helping their patients quit tobacco use. Tobacco dependence consists of both a physical and a psychological dependence. Therefore, the combination of pharmacotherapy with behavior change counseling is recommended. The use of brief Motivational Interviewing (BMI) for tobacco use short interventions in the dental practice appears to be suitable. Nicotine replacement therapy (NRT) is the treatment of choice for the dental practice team because both Varenicline and Bupropion SR have to be prescribed by physicians. PMID- 17425243 TI - Toward specifying the nature of the relationship between expressed emotion and schizophrenic relapse: the utility of curvilinear models. AB - Although expressed emotion (EE) has been demonstrated to be associated with the course of schizophrenia, the nature of this relationship is unclear. This study proposes that testing for curvilinear relationships can identify the specific nature of the relationships between EE indices and relapse. The utility of curvilinear models was explored through a reanalysis of data from a prior study of EE among Mexican-Americans (Karno et al., 1987). The results suggest that the relationship between the EE index of emotional overinvolvement (EOI) and relapse is curvilinear and that high levels of EOI may exert a toxic effect on course of illness whereas medium levels of EOI may be protective. The relationship between the EE index of warmth and relapse is also curvilinear and high levels of warmth appear to exert a protective effect on the course of illness. The role of culture is explored in explaining the specific manner in which EE relates to relapse among Mexican-Americans. PMID- 17425244 TI - Structure analysis of Antonovsky's sense of coherence from an epidemiological mental health survey with a brief nine-item sense of coherence scale. AB - Antonovsky has proposed the sense of coherence (SOC) as a crucial factor that protects against symptoms of mental disease. A central issue in research on the SOC construct is whether this is most appropriately considered as one-dimensional or three-dimensional with comprehensibility (C), manageability (Ma) and meaningfulness (Me) as separate dimensions. In this paper we address this issue by means of confirmatory factor analysis of a shortened nine-item version of Antonovsky's original 29- and 13-items Sense of Coherence Scale (SOCS-29), using epidemiological data from a mental health survey of adults in local communities (N = 1,062). In addition to analysing the internal structure of the SOC items, we examine the association between estimated factor scores and variables expected to be statistically related to SOC. Goodness-of-fit indices were very good for the three-factor model but acceptable even for the one-factor model. In the three factor model, however, the factors were found to be very highly or, with regard to Ma and C, even perfectly correlated. Moreover, the factor scores had very similar correlations with measures of psychological wellbeing, depression and anxiety and they are also very similarly related to age (and none of them are related to gender). We therefore conclude in favour of a one-factor model. PMID- 17425245 TI - Psychometric evaluation of the Spanish version of CONNECT: a measure of continuity of care in mental health services. AB - This article provides the results of the psychometric testing of the Spanish version of CONNECT(-S), a measure of continuity of care in mental health services. CONNECT-S is a multidimensional measure designed for use with seriously mentally ill respondents. Consisting of 12 scales and one single-item indicator, it addresses qualities of interaction in current relationships between mental health service providers and consumers in five conceptual domains: (1) practitioner knowledge of their clients, (2) creating flexibility, (3) practitioner availability, (4) practitioner co-ordination, and (5) smoothing transitions. One-hundred-and-fifty participants took part in the study. Participants were recruited from mental health outpatient clinics in both the Puerto Rican (n = 109) and the San Antonio (n = 41) samples. Internal consistency for scales in a combined site estimate ranged from 0.68 to 0.96. Test-retest reliability ranged from fair to substantial in all but one scale. Concurrent validity hypotheses based on a priori predictions were mostly supported. The Spanish translation and adaptation of CONNECT-S provided sound psychometric results across both sites. CONNECT-S addresses the gap in measurement of continuity of care for the two largest US Latino subgroups, Mexican Americans and Puerto Ricans; and provides an encouraging starting point for a measure that is both relevant and culturally sensitive. PMID- 17425246 TI - Reporting practices of dropouts in psychological research using a wait-list control: current state and suggestions for improvement. AB - Reporting practices regarding dropouts in wait-list control studies hold great importance for the ability to replicate, generalize, and draw conclusions from research. This concern is applicable to all psychological research utilizing wait list controls, regardless of purpose of research (e.g., treatment outcome). The current study assessed the present state of reporting practices in this type of experimental design and discussed the limitations and implications of the insufficient reporting found. 171 articles from psychology journals utilizing wait-list control design were surveyed regarding the reporting of the number of dropouts from the wait-list control and experimental conditions, characteristics and assessment scores of the dropouts, and total dropouts. Variables that are crucial to interpreting research findings are not consistently reported. Additionally, journal impact factor and year of publication were positively correlated with the adequacy of reporting. Consistencies with previous findings were noted, and suggestions for remedying the reporting inadequacies were discussed. PMID- 17425247 TI - Sticking the quantum leap. Modernizing older networks poses formidable IT challenges. Fifty-year-old sharp memorial hospital needed an ultramodern solution to stave off impending obsolescence. PMID- 17425248 TI - Out with the old, in with the new. Virtual item files can streamline ordering, lower costs and afford caregivers more time with patients than current supply data management technology. Should healthcare providers embrace them? PMID- 17425249 TI - Remote Control Training. A bicoastal senior living enterprise reduces training and help desk costs and time with old and new software. PMID- 17425250 TI - Business continuity by design. Don't let a disaster impair your facility's performance. PMID- 17425252 TI - From a distance: saving lives through remote care. A combination of eICU technology, intensivists and in-house staffing delivers significant improvements in ICU patient care. PMID- 17425251 TI - Signed, sealed and indelible. Electronic lab notebooks help a worldwide biopharmaceutical research firm protect intellectual property while reducing paperwork. PMID- 17425253 TI - Better patient care through video interpretation. A New Jersey hospital uses teleconferencing tools and interpreters to break down patient language and hearing barriers. PMID- 17425254 TI - Safeguarding and monitoring Data transmission. A Honolulu-based healthcare enterprise safeguards against confidential data leaks. PMID- 17425255 TI - The keys to the kingdom. How can healthcare organizations grant access to clinical data and applications, while protecting a patient's privacy? For one integrated health system, automated provisioning held the key. PMID- 17425256 TI - When consumers drive, physicians don't have to get taken for a ride. PMID- 17425257 TI - Fate of infectious salmon anaemia virus (ISAV) in experimentally challenged blue mussels Mytilus edulis. AB - In order to investigate the potential role of blue mussels Mytilus edulis as a vector of the fish pathogenic infectious salmon anaemia virus (ISAV), we developed an experimental bioaccumulation system in which mussels can accumulate virus during normal filtration. Detection of virus in mussels was performed by means of real-time RT-PCR. ISAV-RNA was detected in the mussels until 72 h post challenge. Hepatopancreas homogenate from experimentally challenged mussels was injected into salmon. All the fish injected with homogenate prepared immediately after accumulations were strongly ISAV positive 4 wk post-challenge. In the group injected with homogenate prepared 24 h after the challenge, 1 fish out of 25 was weakly ISAV positive. All of the fish that were challenged with mussel homogenate prepared 96 h after accumulation were ISAV negative. Mussels sampled from a tank with experimentally infected salmon demonstrating clinical signs consistent with ISA (infectious salmon anaemia) and mussels collected on net pen cages during ISA outbreaks in Atlantic salmon were all ISAV negative. The results indicate that the ISAV is rapidly inactivated in mussels and that mussels are not a likely reservoir host or vector for ISAV. PMID- 17425258 TI - Retreat sites of rain forest stream frogs are not a reservoir for Batrachochytrium dendrobatidis in northern Queensland, Australia. AB - Chytridiomycosis is a potentially fatal disease of amphibians caused by Batrachochytrium dendrobatidis, and is implicated in declines and extinctions of amphibian populations and species around the world. To cause local host extinction, a disease organism must persist at low host densities. One mechanism that could facilitate this is the ability to persist in the environment. In the laboratory, B. dendrobatidis spreads by both frog-to-frog and environment-to-frog transmission, and can persist on a number of biotic substrates. In the field, B. dendrobatidis has been detected on environmental samples taken during an epidemic, but it is not known if it persists in the environment when endemic. Retreat sites of 2 species of Australian rain forest stream frogs Litoria lesueuri and L. nannotis were sampled 0 to 3 d after occupation during the wet and dry seasons in northern Queensland, Australia, where chytridiomycosis has been endemic for at least 10 yr. The intensity and prevalence of infection in frogs during sampling were comparatively low compared with epidemics. Diagnostic quantitative polymerase chain reaction did not detect B. dendrobatidis in any retreat site samples. It thus appears that retreat sites are not a major environmental source of infection when B. dendrobatidis occurs at low prevalence and intensity on frogs. This suggests that control efforts may not need to eliminate the organism from the environment, at least when prevalence and intensity of infection are low in frogs. Simply treating hosts may be effective at controlling the disease in the wild. PMID- 17425259 TI - Additional disinfectants effective against the amphibian chytrid fungus Batrachochytrium dendrobatidis. AB - Chytridiomycosis, a disease contributing to amphibian declines worldwide, is caused by the fungus Batrachochytrium dendrobatidis. Identifying efficient and practical disinfectants effective against B. dendrobatidis is important to reduce the spread of the disease both in the wild and captivity. Previous studies identified a range of suitable disinfectant strategies. We evaluated the suitability of 3 additional disinfectants: two of these (TriGene Virucidal Disinfectant Cleaner and F10 Super Concentrate Disinfectant) are mixtures of chemicals and one (Betadine Antiseptic Liquid) contains a single active ingredient, povidone iodine. The disinfectants were tested using a range of concentrations for 1,5 and 10 min to determine their ability to kill B. dendrobatidis in vitro. The measure of effectiveness was 100% kill of zoosporangia grown in multiwell plates. All disinfectants had a 100% efficacy at concentrations recommended by the manufacturers. The lowest concentrations capable of 100% kill after exposure for 1 min were 0.1 ml l(-1) for TriGene, 0.33 ml l(-1) for F10 and 100 ml l(-1) for Betadine. TriGene is the most effective disinfectant yet to be found, and both TriGene and F10 are more effective than various disinfectants tested in previous studies. TriGene and F10 are considered suitable for use in the field, as only small amounts of concentrate are needed. PMID- 17425260 TI - Release method and anatomical hook location: effects on short-term mortality of angler-caught Acanthopagrus australis and Argyrosomus japonicus. AB - One field and 3 aquaria experiments were done to quantify the short-term mortality of yellowfin bream Acanthopagrus australis and mulloway Argyrosomus japonicus after being angled and subjected to 3 general handling treatments. Anglers were supplied with identical J-type hooks and asked to handle hooked fish by either (1) physically removing the hook or (2) cutting the line (5 cm from the mouth of the fish) and leaving the hook in. Some hooked A. japonicus were subjected to a third handling treatment where the line was cut underwater without exposing the fish to air. Technical and biological data were collected before all fish were released into sea cages and monitored for 5 d. Control fish were seined and similarly caged and monitored. Concentrations of plasma glucose and cortisol were collected from a sample of fish on the first and last day of the experiments. Significant predictors of mortality for both species involved the presence of blood at the mouth and an interaction between anatomical hook location and hook removal. A. australis and A. japonicus that had their ingested hooks removed experienced the greatest mortalities (87.5 and 72.7%, respectively). Typically, these fish suffered damage to their oesophagus, stomach wall and vital organs. Mortality rates of A. australis and A. japonicus were significantly decreased to 1.7 and 16%, respectively, when they were released with their lines cut, with some of these fish free of hooks after 5 d. In contrast, few mortalities occurred in either species when the hooks were removed or the lines cut on mouth-hooked fish or in A. japonicus when it was released with no air exposure. For A. australis, the field- and aquaria-based experiments provided comparable results in terms of identifying treatment-specific effects, but there were potential biases in rates of hook ingestion. Irrespective of the treatment of fish, all experiments caused physiological changes measured as elevations in either plasma cortisol or glucose. We concluded that anglers should cut the line from hook-ingested A. australis and A. japonicus, but remove the hook from mouth-hooked individuals to prevent subsequent ingestion. Further research is required to examine the longer-term consequences of these handling practices on fish health. PMID- 17425261 TI - Ingestion and ejection of hooks: effects on long-term health and mortality of angler-caught yellowfin bream Acanthopagrus australis. AB - Ninety juvenile yellowfin bream Acanthopagrus australis were angled from holding tanks, allowed to ingest nickel-plated, carbon-steel J-hooks and released (with their lines cut) into individual experimental tanks during 2 experiments in order to assess their (1) long-term (up to 105 d) health, mortality and rate of hook ejection and (2) short- and medium-term (< 42 d) temporal changes in health during hook ingestion. Equal numbers of control fish were scooped from holding tanks and similarly monitored in experimental tanks. Of 20 hook-ingested fish released during Expt 1, 3 died within 8 d, providing a non-significant mortality of 15%. Between Day 6 and Day 56 post-release, 13 of the surviving individuals ejected their hooks, which were typically oxidized to about 94% of their original weight and often broken into 2 pieces. At Day 105, there were no significant differences between the 20 control and 17 hook-ingested/-ejected fish in terms of their ability to digest and assimilate food (measured as changes in apparent digestibility coefficients), stress (measured as concentrations of plasma cortisol and glucose) or of morphological parameters that included weight (Wt) and maximum height (MH), maximum width (MW) and maximum girth (MG). During Expt 2, 3 individuals that still contained ingested hooks and 3 controls were sampled on each of 9 occasions between Day 3 and Day 42 post-release. All fish were sampled for blood cortisol and glucose and were then euthanized before being weighed and measured for total length (TL), MH, MW and MG. Hook-ingested individuals were also X-rayed to determine the position and orientation of hooks. There were no significant differences in plasma glucose between hook-ingested and control fish. Irrespective of the treatment of fish, concentrations of cortisol were elevated on some sampling occasions, indicating variable, acute stress. The MH and MG of fish were not significantly different between groups. Significant differences were detected for MG and Wt, with hook-ingested fish having weights similar to those of the control fish but a relatively greater MW (owing to stomach distension from ingested hooks) until 2 wk post-release, after which both morphological parameters generally declined. There was no significant temporal progression of hooks in the stomach of treatment fish; however, some hooks reorientated to positions that may have precluded passage along the digestive tract. We conclude that, for the J-hooks examined, cutting the line is an appropriate strategy that results in the greater majority of released hook ingested yellowfin bream surviving with minimal negative long-term effects. PMID- 17425262 TI - Identification of potential vaccine target antigens by immunoproteomic analysis of a virulent and a non-virulent strain of the fish pathogen Flavobacterium psychrophilum. AB - Flavobacterium psychrophilum is the etiological agent of bacterial coldwater disease (CWD) and rainbow trout fry syndrome (RTFS). To identify antigens associated with virulence or host immunity, we compared total and immunogenic proteins of cellular and extracellular products (ECP) between a virulent (CSF-259 93) and non-virulent (ATCC 49418) strain of F. psychrophilum. One-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis of total cellular proteins revealed only minor differences between the strains; however, separation of ECP showed that proteins were differentially expressed. Western blot analysis using rainbow trout (Oncorhynchus mykiss) anti-CSF-259-93 sera showed greater reactivity to proteins of the virulent strain, including many > 50 kDa. Further analysis by 2-dimensional electrophoresis (2DE) identified numerous differences between the strains. Western blot analysis combined with 2DE identified several immunogenic proteins that reacted with the antisera and were shared between the 2 strains. However, at least 15 immunogenic proteins appeared to be unique to the virulent strain, while 4 such proteins were identified in the non-virulent strain; 8 proteins unique to the virulent strain and 6 shared proteins were further analyzed for identification by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis. Of these, 3 immunogenic proteins (heat shock proteins HSP 60 and HSP 70) and 2 other proteins (ATP synthase and thermolysin) were conclusively identified. The 2 highly immunogenic heat shock proteins were shown to share extensive homology with heat shock proteins of related bacteria. This approach for antigen identification may provide a basis for targeted vaccine development against CWD and RTFS. PMID- 17425263 TI - Evaluation of a cohabitation challenge model in immunization trials for channel catfish Ictalurus punctatus against Ichthyophthirius multifiliis. AB - Calcein marking and cohabitation challenges have not been investigated in fish parasite research. This study evaluated a cohabitation challenge method in immunization trials against Ichthyophthirius multifiliis (Ich) using calcein, a fluorescent dye, to mark channel catfish Ictalurus punctatus (Rafinesque). Fish were marked by calcein immersion at 0, 500, and 1500 mg l(-1), and then challenged with 15 000 theronts fish(-1). No difference was noted in fish infection levels, mortality, and mean days to death (MDD) caused by Ich between unmarked and marked fish or between fish marked with high (1500 mg l(-1)) and low (500 mg l(-1)) concentrations of calcein. After ensuring that calcein marking had no effect on the susceptibility of fish to Ich theronts, 2 immunization trials were conducted to evaluate the cohabitation challenge model using calcein-marked catfish. Fish mortality, relative percent survival (RPS), and MDD were compared between cohabitation-challenged fish and fish challenged by non-cohabitation. No significant difference was observed in RPS for cohabitation-challenged fish and fish challenged by non-cohabitation. A cohabitation challenge can be used as an alternative challenge method in parasite studies, since it closely mimics natural exposure. PMID- 17425264 TI - Phylogeny of Neoparamoeba strains isolated from marine fish and invertebrates as inferred from SSU rDNA sequences. AB - We characterised 9 strains selected from primary isolates referable to Paramoeba/Neoparamoeba spp. Based on ultrastructural study, 5 strains isolated from fish (amoebic gill disease [AGD]-affected Atlantic salmon and dead southern bluefin tuna), 1 strain from netting of a floating sea cage and 3 strains isolated from invertebrates (sea urchins and crab) were assigned to the genus Neoparamoeba Page, 1987. Phylogenetic analyses based on SSU rDNA sequences revealed affiliations of newly introduced and previously analysed Neoparamoeba strains. Three strains from the invertebrates and 2 out of 3 strains from gills of southern bluefin tunas were members of the N. branchiphila clade, while the remaining, fish-isolated strains, as well as the fish cage strain, clustered within the clade of N. pemaquidensis. These findings and previous reports point to the possibility that N. pemaquidensis and N. branchiphila can affect both fish and invertebrates. A new potential fish host, southern bluefin tuna, was included in the list of farmed fish endangered by N. branchiphila. The sequence of P. eilhardi (Culture Collection of Algae and Protozoa [CCAP] strain 1560/2) appeared in all analyses among sequences of strain representatives of Neoparamoeba species, in a position well supported by bootstrap value, Bremer index and Bayesian posterior probability. Our research shows that isolation of additional strains from invertebrates and further analyses of relations between molecular data and morphological characters of the genera Paramoeba and Neoparamoeba are required. This complexity needs to be considered when attempting to define molecular markers for identification of Paramoeba/Neoparamoeba species in tissues of fish and invertebrates. PMID- 17425265 TI - Characterization of a rediscovered haplosporidian parasite from cultured Penaeus vannamei. AB - Mortalities of Penaeus vannamei, cultured in ponds in Belize, Central America, began during the last part of the grow-out cycle during the cold weather months from September 2004 through February 2005. Tissue squashes of infected hepatopancreata and histological examination of infected shrimp revealed that the mortalities might have been caused by an endoparasite. To confirm the diagnosis, DNA was extracted from ethanol preserved hepatopancreata and the small-subunit rRNA gene was sequenced. The 1838 bp sequence was novel and phylogenetic analysis placed the P. vannamei parasite within the phylum Haplosporidia as a sister taxon to a clade that includes Bonamia and Minchinia species. In situ hybridization was performed using anti-sense DNA probes that were designed to hybridize specifically with the parasite's nucleic acid. This organism presents similar characteristics to those of a haplosporidian that infected cultured P. vannamei imported from Nicaragua into Cuba, as described by Dykova et al. (1988; Fish Dis 11:15-22). PMID- 17425266 TI - Taura syndrome virus in specific pathogen-free Penaeus vannamei originating from Hawaii and in P. vannamei stocks farmed in France? AB - It is the opinion of the authors of the Comment on Do et al. (2006), that those authors incorrectly interpreted their test results, which are more likely the result of mislabeling of samples or within-laboratory contamination, and that the TSV isolates found in Penaeus vannamei in Korea in 2004 and 2005 did not originate from Hawaii as claimed by the authors, but from a country (or countries) in southeast Asia. Finally, we believe that the authors did not follow proper international guidelines, extend a professional courtesy to the supplier of the disputed shrimp sample, nor take a critical approach in interpreting their own data. It is unfortunate that the authors did not follow through with additional testing, or seek a second opinion from an independent laboratory, before implicating shrimp imported from Hawaii as the source of TSV in Korea. PMID- 17425268 TI - Good vibrations. PMID- 17425267 TI - A surgeon on the front line. PMID- 17425269 TI - Yes, we really are what we eat. PMID- 17425270 TI - Ask and you shall receive. PMID- 17425271 TI - Can google make you a better doctor? PMID- 17425272 TI - Going bare? Bad idea. PMID- 17425273 TI - Breaking bad habits. PMID- 17425274 TI - Collections. Learn from the pros. PMID- 17425275 TI - How to ask for copays. PMID- 17425276 TI - "About the size of....". PMID- 17425277 TI - Don't miss these business deductions. PMID- 17425278 TI - Compliance patient autonomy vs P4P imperatives. PMID- 17425279 TI - What about a crackdown on patients? PMID- 17425280 TI - How to get patients on board. PMID- 17425281 TI - An efficient bicomponent TiO2/SnO2 nanofiber photocatalyst fabricated by electrospinning with a side-by-side dual spinneret method. AB - In this communication, we demonstrate that the electrospinning process with a side-by-side dual spinneret can be a simple approach for fabricating bicomponent TiO2/SnO2 nanofibers with controllable heterojunctions. Specifically, both of the TiO2 and SnO2 components in the nanofibers are fully exposed to the surface. This morphology fully utilized the photogenerated holes and electrons during the photocatalytic process, thus leading to a high photocatalytic activity. We believe that this versatile approach can be extended to fabricate other novel high-efficiency bicomponent photocatalysts. PMID- 17425282 TI - Shall we dance? How a multicopper oxidase chooses its electron transfer partner. AB - Multicopper oxidases (MCOs) are encoded in the genomes of Eukarya, Bacteria, and Archea. These proteins are unique in that they contain at least four Cu atom prosthetic groups organized into one each of the three spectral classifications of copper sites in biology: type 1 (T1), type 2 (T2), and binuclear type 3 (T3), where the T2 and T3 sites form a trinuclear Cu cluster. With these four redox active copper sites, the multicopper oxidases catalyze the four-electron (4e(-)) reduction of dioxygen to 2H2O, an activity that they alone share with the terminal heme-containing oxidases. Most MCOs exhibit broad specificity towards organic reductants, while a relatively small number of family members exhibit equally robust activity towards metal ions like Fe(II), Cu(I), and Mn(II) and, thus, are considered metallo-oxidases. This Account analyzes the structure activity features of multicopper oxidases that determine their relative substrate specificity. Since the substrate oxidation step involves an outer-sphere electron transfer from the reductant to the T1Cu site in the protein, the concepts of Marcus theory are applied to unravel the origin of the substrate specificity of the multicopper ferroxidases. PMID- 17425283 TI - Bright oligothiophene N-succinimidyl esters for efficient fluorescent labeling of proteins and oligonucleotides. PMID- 17425284 TI - Validation studies of thermal extraction-GC/MS applied to source emissions aerosols. 1. Semivolatile analyte-nonvolatile matrix interactions. AB - In this work, we develop a novel validation approach for studying how nonvolatile aerosol matrixes of considerably different chemical composition potentially affect the thermal extraction (TE)-GC/MS quantification of a wide range of trace semivolatile organic markers. The nonvolatile matrixes of a set of source emissions aerosols are first operationally isolated by thermally clearing the aerosols of their native semivolatile organic matter. TE-GC/MS analysis is then performed in triplicate on matrixes refortified with multilevel organic compound standard suites. The spiking of empty thermal extraction tubes and blank quartz filters is introduced as experimental control and also allows for the calculation of method detection limits. For the vast majority of organic compounds fortifying the matrixes (e.g., the alkane, alkene, cycloalkane, sterane, and phthalate classes), the analytical bias observed was classified as either minor or nonexistent. Furthermore, compound recoveries were generally highly reproducible, demonstrating relative standard deviations of less than 20%. For a diesel engine exhaust sample, significant matrix effects for the six- and seven-ring polycyclic aromatic hydrocarbons (PAHs) are observed and ascribed to the high proportion of elemental carbon in the sample. Our results suggest that TE-GC/MS may underestimate inhalation exposures to PAHs (with 5 rings or more) in atmospheric aerosols replete with diesel engine exhaust (e.g., near roadways or in polluted urban air). Due to its stability and representativeness, the use of a thermally cleared particulate matter matrix for validation purposes is probably expandable to additional sample pretreatment and instrumental techniques also being applied to quantify organic molecular markers in source and atmospheric aerosols. PMID- 17425285 TI - Automatic in vitro determination of hypochlorous acid scavenging capacity exploiting multisyringe flow injection analysis and chemiluminescence. AB - In the present work, a chemiluminometric automatic flow methodology for the in vitro determination of hypochlorous acid scavenging capacity, under pH and concentration conditions similar to those found in vivo, is proposed. As the pH found in physiological conditions (7.4) and the pH required for the chemiluminescence detection reaction (>10) are different, the multisyringe flow injection analysis features were exploited to perform the in-line reaction of HOCl and the scavenger molecule at physiological pH prior to reaction of the remaining HOCl with luminol at alkaline conditions. These two reactions were carried out in about 3 s, allowing the determination of fast reacting antioxidants, in a time frame closer to in vivo generation of HOCl when compared to previously described methods. The developed method was applied to nonsteroidal antiinflammatory drugs of different chemical families, and positive controls (cysteine, gallic acid, lipoic acid). The HOCl scavenging capacity was evaluated at pH 7.4 and 10.0; different results were found for oxicam derivatives, providing evidence that the pH of in vitro methods should be carefully selected to allow assumptions about putative in vivo effects. PMID- 17425286 TI - Modification of tryptophan and methionine residues is implicated in the oxidative inactivation of surfactant protein B. AB - Exposing BLES (bovine lipid extract surfactant), a clinical surfactant, to reactive oxygen species (ROS) alters surfactant protein B (SP-B), as indicated by Coomassie Blue staining, silver staining, and Western analysis. Hypochlorous acid (HOCl) treatment leads to elevated maximum surface tension (gammamax) and a deterioration in minimum gamma (gammamin) during surface area cycling. Fenton reaction resulted in immediate increases in gammamin and gammamax. Intrinsic fluorescence measurements indicated Fenton, but not HOCl, induced conversion of Trp9 of SP-B to hydroxyTrp (OHTrp), N-formylkynurenine (NFKyn), and kynurenine (Kyn). Electrospray ionization mass spectrometry (ESI-MS) revealed molecular weight alterations consistent with oxidation of Met (HOCl, Fenton) and Trp (Fenton) residues. Oxidative alterations to Met29 and Met65 (HOCl, Fenton) and to Trp9 (OHTrp with HOCL and NFKyn plus Kyn with Fenton) were confirmed by matrix assisted laser desorption mass spectrometry (MALDI-MS) studies on SP-B tryptic fragments. Some Met oxidation was observed with control SP-B. When taken together with captive bubble tensiometer measurements, these studies suggest that Met oxidation of SP-B by HOCl or Fenton interferes with phospholipid respreading during compression-expansion of surfactant films, while Fenton oxidation, which produces more extensive Met oxidation and disruption of the indole ring of Trp9, further abrogated the ability of such films to attain low surface tensions during compression. These studies provide insight into the manner by which ROS generated during acute lung injury and the acute respiratory distress syndrome act to inhibit not only endogenous surfactant but also therapeutic surfactants administered to counteract these conditions. PMID- 17425288 TI - Structural features of target RNA molecules greatly modulate the cleavage efficiency of trans-acting delta ribozymes. AB - The aim of this work was to shed some more light on factors influencing the effectiveness of delta ribozyme cleavage of structured RNA molecules. An oligoribonucleotide that corresponds to the 3'-terminal region X of HCV RNA and yeast tRNAPhe were used as representative RNA targets. Only a few sites susceptible to ribozyme cleavage were identified in these targets using a combinatorial library of ribozyme variants, in which the region responsible for ribozyme-target interaction was randomized. On the other hand, the targets were fairly accessible for binding of complementary oligonucleotides, as was shown by 6-mer DNA libraries and RNase H approach. Moreover, the specifically acting ribozymes cleaved the targets precisely but with unexpectedly modest efficacy. To explain these observations, six model RNA molecules were designed, in which the same seven nucleotide long sequence recognized by the delta ribozyme was always single stranded but was embedded into different RNA structural context. These molecules were cleaved with differentiated rates, and the corresponding k2 values were in the range of 0.91-0.021 min-1; thus they differed almost 50-fold. This clearly shows that cleavage of structured RNAs might be much slower than cleavage of a short unstructured oligoribonucleotide, despite full accessibility of the targeted regions for hybridization. Restricted possibilities of conformational transitions, which are necessary to occur on the cleavage reaction trajectory, seem to be responsible for these differences. Their magnitude, which was evaluated in this work, should be taken into account while considering the use of delta ribozymes for practical applications. PMID- 17425287 TI - Analysis of the gastric H,K ATPase for ion pathways and inhibitor binding sites. AB - New models of the gastric H,K ATPase in the E1K and E2P states are presented as the first structures of a K+ counter-transport P2-type ATPase exhibiting ion entry and exit paths. Homology modeling was first used to generate a starting conformation from the srCa ATPase E2P form (PDB code 1wpg) that contains bound MgADP. Energy minimization of the model showed a conserved adenosine site but nonconserved polyphosphate contacts compared to the srCa ATPase. Molecular dynamics was then employed to expand the luminal entry sufficiently to allow access of the rigid K+ competitive naphthyridine inhibitor, Byk99, to its binding site within the membrane domain. The new E2P model had increased separation between transmembrane segments M3 through M8, and addition of water in this space showed not only an inhibitor entry path to the luminal vestibule but also a channel leading to the ion binding site. Addition of K+ to the hydrated channel with molecular dynamics modeling of ion movement identified a pathway for K+ from the lumen to the ion binding site to give E2K. A K+ exit path to the cytoplasm operating during the normal catalytic cycle is also proposed on the basis of an E1K homology model derived from the E12Ca2+ form of the srCa ATPase (PDB code 1su4). Autodock analyses of the new E2P model now correctly discriminate between high- and low-affinity K+ competitive inhibitors. Finally, the expanded luminal vestibule of the E2P model explains high-affinity ouabain binding in a mutant of the H,K ATPase [Qiu et al. (2005) J. Biol. Chem. 280, 32349-32355]. PMID- 17425289 TI - Kinetic mechanism of enterococcal aminoglycoside phosphotransferase 2"-Ib. AB - The major mechanism of resistance to aminoglycosides in clinical bacterial isolates is the covalent modification of these antibiotics by enzymes produced by the bacteria. Aminoglycoside 2''-Ib phosphotransferase [APH(2'')-Ib] produces resistance to several clinically important aminoglycosides in both Gram-positive and Gram-negative bacteria. Nuclear magnetic resonance analysis of the product of kanamycin A phosphorylation revealed that modification occurs at the 2''-hydroxyl of the aminoglycoside. APH(2'')-Ib phosphorylates 4,6-disubstituted aminoglycosides with kcat/Km values of 10(5)-10(7) M-1 s-1, while 4,5 disubstituted antibiotics are not substrates for the enzyme. Initial velocity studies demonstrate that APH(2'')-Ib operates by a sequential mechanism. Product and dead-end inhibition patterns indicate that binding of aminoglycoside antibiotic and ATP occurs in a random manner. These data, together with the results of solvent isotope and viscosity effect studies, demonstrate that APH(2'')-Ib follows the random Bi-Bi kinetic mechanism and substrate binding and/or product release could limit the rate of reaction. PMID- 17425290 TI - A gas-phase kinetic study of the reaction between bromine monoxide and methylperoxy radicals at atmospheric temperatures. AB - The rate constant of the reaction of BrO with CH(3)O(2) was determined to be k1 = (6.2 +/- 2.5) x 10(-12) cm3 molecule(-1) s(-1) at 298 K and 100-200 Torr of O2 diluent. Quoted uncertainty was two standard deviations. No significant pressure dependence of the rate constants was observed at 100-200 Torr total pressure of N2 or O2 diluents. Temperature dependence of the rate constants was further investigated over the range 233-333 K, and an Arrhenius type expression was obtained for k1 = 4.6 x 10(-13) exp[(798 +/- 76)/T] cm3 molecule(-1) s(-1). The product branching ratios were evaluated and the atmospheric implications were discussed. PMID- 17425292 TI - Blue-shifting intramolecular C-H...O interactions. AB - Two model systems, 3-methylacroleine and 3-(difluoromethyl)acroleine, are investigated computationally with respect to the character of the C-H...O interaction in their chelate-type (ZZ) conformers. By selecting the appropriate reference conformers, the C-H...O interaction is shown to result in the increase of the C-H stretching frequency (i.e., in the blue shift of the C-H stretching band). This is accompanied by the shortening of the C-H bond distance as compared to its values in reference molecules. Parallel to calculations of the C-H bond distance and stretching frequency, the energy contribution of the C-H...O interaction to the total energy (i.e., the energy associated with the C-H...O contact) is evaluated by using the methods proposed recently for the estimation of the energies of intramolecular hydrogen bonds. It is found that the C-H...O contact in the chelate-type forms of 3-methylacroleine and 3 (difluoromethyl)acroleine corresponds to the negative energy contribution and is repulsive. It is concluded that, despite the stability of the ZZ conformers of the two molecules and their deceptive structural shape, no hydrogen bond in the usual sense is formed between the C-H bond and the lone pair donor. The results are interpreted in terms of the steric compression, which leads to the dominance of the valence repulsion contribution in the C-H...O contact. This mechanism suggests that blue-shifting intramolecular interactions should not be that uncommon, although their recognition requires a careful consideration of the reference system. PMID- 17425291 TI - Probing P-H+-P hydrogen bonds: structures, binding energies, and spin-spin coupling constants. AB - Ab initio MP2/aug'-cc-pVTZ calculations have been performed to determine the structures and binding energies of 22 open and 3 cyclic complexes formed from the sp2 [H(2)C=PH and HP=PH (cis and trans)] and sp3 [PH2(CH3) and PH3] hybridized phosphorus bases and their corresponding protonated ions. EOM-CCSD calculations have been carried out to obtain (31)P-(31)P and (31)P-(1)H coupling constants across P-H+-P hydrogen bonds. Two equilibrium structures with essentially linear hydrogen bonds have been found along the proton-transfer coordinate, except for complexes with P(CH3)H3+ as the proton donor to the sp2 bases. Although the isomer having the conjugate acid of the stronger base as the proton donor lies lower on the potential energy surface, it has a smaller binding energy relative to the corresponding isolated monomers than the isomer with the conjugate acid of the weaker base as the donor. The hydrogen bond of the latter has increased proton-shared character. All of the complexes are stabilized by traditional hydrogen bonds, as indicated by positive values of the reduced coupling constants (2h)K(P-P) and (1)K(P-H), and negative values of (1h)K(H-P). (2h)J(P-P) correlates with the P-P distance, a correlation determined primarily by the nature of the proton donor. For open complexes, (1)J(P-H) always increases relative to the isolated monomer, while (1h)J(H-P) is relatively small and negative. (2h)J(P-P) values are quite large in open complexes, but are much smaller in cyclic complexes in which the P-H+-P hydrogen bonds are nonlinear. Thus, experimental measurements of (2h)J(P-P) should be able to differentiate between open and cyclic complexes. PMID- 17425293 TI - Laser-induced fluorescence spectrum of 3-vinyl-1H-indene. AB - The laser-induced fluorescence spectrum of 3-vinyl-1H-indene was recorded between 33,000 and 33,800 cm(-1). An origin band was observed at 33,455 cm(-1) along with several low-frequency modes. With the aid of density functional theory and configuration interaction calculations, the electronic transition was assigned as S1 <-- S0 and the short progression in an 80 cm(-1) mode was identified as a vinyl group torsion. Theoretical, spectroscopic, and thermochemical considerations suggest that the 3-vinyl-1H-indene spectrum results from excitation from both conformational isomers with the vinyl and indene double bonds in trans and cis arrangements. The results are discussed in the context of the identification of species arising from the discharge of benzene in argon. PMID- 17425294 TI - Conformation of L-tyrosine studied by fluorescence-detected UV-UV and IR-UV double-resonance spectroscopy. AB - The laser-induced fluorescence spectrum of jet-cooled L-tyrosine exhibits more than 20 vibronic bands in the 35450-35750 cm(-1) region. We attribute these bands to eight conformers by using results of UV-UV hole-burning spectroscopy. These isomers are classified into four groups; each group consists of two rotational isomers that have a similar side-chain conformation but different orientations of the phenolic OH. The splitting of band origins of rotational isomers is 31, 21, 5, and 0 cm(-1) for these groups. IR-UV spectra suggest that conformers belonging to two of the four groups have an intramolecular OH...N hydrogen bond between the COOH and NH2 groups. By comparing experimental and theoretical results of L tyrosine with those of L-phenylalanine, we propose probable conformers of L tyrosine. PMID- 17425295 TI - Quantitative structure-activity relationship of peptides binding to the class II major histocompatibility complex molecule Aq associated with autoimmune arthritis. AB - Presentation of (glyco)peptides by the class II major histocompatibility complex molecule Aq to T cells plays a central role in collagen-induced arthritis, an animal model for the autoimmune disease rheumatoid arthritis. A peptide library was designed using statistical molecular design in amino acid space in which five positions in the minimal mouse collagen type II binding epitope CII260-267 were varied. A substantially reduced peptide library of 24 peptides with diverse and representative molecular characteristics was selected, synthesized, and evaluated for the binding strength to Aq. A multivariate QSAR model was established by correlating calculated descriptors, compressed to its principle properties, with the binding data using partial least-square regression. The model was successfully validated by an external test set. Interpretation of the model provided a molecular property binding motif for peptides interacting with Aq. The information may be useful in future research directed toward new treatments of rheumatoid arthritis. PMID- 17425296 TI - 1,4-Dihydroindeno[1,2-c]pyrazoles with acetylenic side chains as novel and potent multitargeted receptor tyrosine kinase inhibitors with low affinity for the hERG ion channel. AB - The synthesis of a novel series of 1,4-dihydroindeno[1,2-c]pyrazoles with acetylene-type side chains is described. Optimization of those compounds as KDR kinase inhibitors identified 8, which displayed an oral activity in an estradiol induced murine uterine edema model (ED50 = 3 mg/kg) superior to Sutent (ED50 = 9 mg/kg) and showed potent antitumor efficacy in an MX-1 human breast carcinoma xenograft tumor growth model (tumor growth inhibition = 90% at 25 mg/kg.day po). The compound was docked into a homology model of the homo-tetrameric pore domain of the hERG potassium channel to identify strategies to improve its cardiac safety profile. Systematic interruption of key binding interactions between 8 and Phe656, Tyr652, and Ser624 yielded 90, which only showed an IC50 of 11.6 microM in the hERG patch clamp assay. The selectivity profile for 8 and 90 revealed that both compounds are multitargeted receptor tyrosine kinase inhibitors with low nanomolar potencies against the members of the VEGFR and PDGFR kinase subfamilies. PMID- 17425298 TI - Virtual screening for novel openers of pancreatic K(ATP) channels. AB - Ligand-based virtual screening approaches were applied to search for new chemotype KCOs activating Kir6.2/SUR1 KATP channels. A total of 65 208 commercially available compounds, extracted from the ZINC archive, served as database for screening. In a first step, pharmacokinetic filtering via VolSurf reduced the initial database to 1913 compounds. Afterward, six molecules were selected as templates for similarity searches: similarity scores, obtained toward these templates, were calculated with the GRIND, FLAP, and TOPP approaches, which differently encode structural information into potential pharmacophores. In this way, we obtained 32 hit candidates, 16 via GRIND and eight each via FLAP and TOPP. For biological testing of the hit candidates, their effects on membrane potentials in HEK 293 cells expressing Kir6.2/SUR1 were studied. GRIND, FLAP, and TOPP all yielded hits, but no method top-ranked all the actives. Thus, parallel application of different approaches probably improves hit detection. PMID- 17425297 TI - Structure-based design and synthesis of N(omega)-nitro-L-arginine-containing peptidomimetics as selective inhibitors of neuronal nitric oxide synthase. Displacement of the heme structural water. AB - The neuronal isoform of nitric oxide synthase (nNOS), the enzyme responsible for the production of nitric oxide in the central nervous system, represents an attractive target for the treatment of various neurodegenerative disorders. X-ray crystal structures of complexes of nNOS with two nNOS-selective inhibitors, (4S) N-{4-amino-5-[(2-aminoethylamino]pentyl}-N'-nitroguanidine (1) and 4-N-(Nomega nitro-l-argininyl)-trans-4-amino-l-proline amide (2), led to the discovery of a conserved structural water molecule that was hydrogen bonded between the two heme propionates and the inhibitors (Figure 2). On the basis of this observation, we hypothesized that by attaching a hydrogen bond donor group to the amide nitrogen of 2 or to the secondary amine nitrogen of 1, the inhibitor molecules could displace the structural water molecule and obtain a direct interaction with the heme cofactor. To test this hypothesis, peptidomimetic analogues 3-5, which have either an N-hydroxyl (3 and 5) or N-amino (4) donor group, were designed and synthesized. X-ray crystal structures of nNOS with inhibitors 3 and 5 bound verified that the N-hydroxyl group had, indeed, displaced the structural water molecule and provided a direct interaction with the heme propionate moiety (Figures 5 and 6). Surprisingly, in vitro activity assay results indicated that the addition of a hydroxyl group (3) only increased the potency slightly against the neuronal isoform over the parent compound (1). Rationalizations for the small increase in potency are consistent with other changes in the crystal structures. PMID- 17425299 TI - Novel selective neuropeptide Y2 receptor PEGylated peptide agonists reduce food intake and body weight in mice. AB - Selective activation of the NPY2 receptor to suppress appetite provides an approach to obesity management. Selective NPY2 PEGylated peptide agonists are described that consist of a peptide core corresponding to residues 25-36 of PYY and a nonpeptidic moiety at the peptide N-terminus that contributes to in vitro potency and in vivo efficacy and provides a PEGylation site. The lead peptide elicits a dose-dependent reduction of food intake in lean mice and of food intake, body weight, and fat mass in DIO mice. PMID- 17425300 TI - Identifying promising compounds in drug discovery: genetic algorithms and some new statistical techniques. AB - Throughout the drug discovery process, discovery teams are compelled to use statistics for making decisions using data from a variety of inputs. For instance, teams are asked to prioritize compounds for subsequent stages of the drug discovery process, given results from multiple screens. To assist in the prioritization process, we propose a desirability function to account for a priori scientific knowledge; compounds can then be prioritized based on their desirability scores. In addition to identifying existing desirable compounds, teams often use prior knowledge to suggest new, potentially promising compounds to be created in the laboratory. Because the chemistry space to search can be dauntingly large, we propose the sequential elimination of level combinations (SELC) method for identifying new optimal compounds. We illustrate this method on a combinatorial chemistry example. PMID- 17425301 TI - Exploration of rate-limiting conformational state for 5-[(7-chloro-4 quinolinyl)amino]-3-[(alkylamino)methyl][1,1'-biphenyl]-2-ols and N(omega)-oxides (tebuquine analogues) for antimalarial activity using molecular shape analysis and molecular field analysis studies. AB - Tebuquine is a 4-aminoquinoline that shows significantly more potency as an antimalarial than amodiaquine and chloroquine both in vitro and in vivo. To explore the conformation in the rate-limiting step and to elucidate pharmacophoric properties of tebuquine-related analogues, molecular shape analysis (MSA) along with molecular field analysis (MFA) methods were applied on a series of 5-[(7-chloro-4-quinolinyl)amino]-3-[(alkylamino)methyl][1,1' biphenyl]-2-ol analogues and their Nomega-oxides possessing antimalarial activity. The study was performed using 45 compounds in which 37 molecules were taken as a training set for the derivation of the 3D quantitative structure activity relationship models and eight molecules were kept as a test set to evaluate the predictive ability of the derived models. Both methods were analyzed in terms of their predictive abilities and produced comparable results with good conventional and cross-validated r2 values (0.908 and 0.886, respectively, for the MFA model and 0.846 and 0.812, respectively, for the MSA model). In external data set prediction, the MSA model scored much better than MFA. Steric, electrostatic, and hydrogen-bond donor/acceptor fields of molecules were found to be relevant descriptors for structure-activity relationships. The inclusion of polar solvent-accessible charged surface area and spatial descriptors in the MSA model generation resulted in a model with significant predictive ability for the test set molecules. This indicates the importance of the orientation of conformationally favored molecules inside the receptor site and solvation of the charged surfaces of the molecule by a polar solvent for the activity of the molecule. The results provided the appropriate tools for predicting the affinity of related compounds using a ligand-based approach, and for guiding the design and synthesis of novel and more potent antimalarial agents. PMID- 17425302 TI - A density functional theory study of the reaction of the biomimetic iron(II) complex of a tetradentate bispidine ligand with H2O2. AB - Three pathways for the reaction of bispidine-iron(II) complexes (where bispidine is a rigid tetradentate amine/pyridine ligand) with H2O2 have been studied by DFT calculations. For all oxidation states the high-spin and low-spin (intermediate spin) forms have been optimized, and the computed data have been compared with the readily available experimental results. It is concluded that there is a direct conversion of the bispidine-iron(II)-hydrogen peroxide complex to the corresponding iron(IV)-dihydroxo compound, which is a novel possible oxidant for the dihydroxylation of olefins. PMID- 17425303 TI - Photomagnetic properties of an iron(II) low-spin complex with an unusually long lived metastable LIESST state. AB - A comprehensive study of the photomagnetic behavior of the [Fe(L222N5)(CN)2].H2O complex has been carried out. This complex is characterized by a low-spin (LS) iron(II)-metal center up to 400 K and exhibits at 10 K the well-known Light Induced Excited Spin State Trapping (LIESST) effect. The critical LIESST temperature (T(LIESST)) has been measured to be 105 K. The kinetics of the transition from the metastable high-spin (HS) state to the low-spin state have been determined and used for reproducing the experimental T(LIESST) curve. This study represents a second example of a fully low-spin iron(II)-metal complex up to 400 K, which can be photoexcited at low temperature with an atypical long lived metastable HS state. This underlines the preponderant role of the inner coordination sphere for stabilizing the lifetime of the photoinduced HS state. PMID- 17425304 TI - Oligonuclear copper complexes of a bioinspired pyrazolate-bridging ligand: synthesis, structures, and equilibria in solution. AB - The synthesis of a new bioinspired dinucleating ligand scaffold based on a bridging pyrazolate with appended bis[2-(1-methylimidazolyl)methyl]aminomethyl chelate arms is reported. This ligand forms very stable copper complexes, and a series of different species is present in solution depending on the pH. Interconversions between these solution species are tracked and characterized spectroscopically, and X-ray crystallographic structures of three distinct complexes that correspond to the species present in solution from acidic to basic pH have been determined. Overall, this provides a comprehensive picture of the copper coordination chemistry of the new ligand system. Alterations in the protonation state are accompanied by changes in nuclearity and pyrazolate binding, which cause pronounced changes in color and magnetic properties. Antiferromagnetic coupling between the copper(II) ions is switched on or off depending on the pyrazole binding mode. PMID- 17425305 TI - Alkaline earth metal complexes of a phosphine-borane-stabilized carbanion: synthesis, structures, and stabilities. AB - The reaction between either MgI2 or CaI2 and 2 equiv of [(Me3Si)2{Me2(H3B)P}C]K (2) in toluene gives the corresponding organo-alkaline earth metal compounds [(Me3Si)2{Me2(H3B)P}C]2M in moderate to good yields [M = Mg (3), Ca (4)]. Compound 3 crystallizes solvent-free, whereas X-ray quality crystals of 4 could not be obtained in the absence of coordinating solvents; crystallization of 4 from cold methylcyclohexane/THF gives the solvate [(Me3Si)2{Me2(H3B)P}C]2Ca(THF)4 (4a). The corresponding heavier alkaline earth metal complexes [(Me3Si)2{Me2(H3B)P}C]2M(THF)5 [M = Sr (7), Ba (8)] are obtained from the reaction between MI2 and 2 equiv of 2 in THF, followed by recrystallization from cold methylcyclohexane/THF. Compound 3 degrades over a period of several weeks at room-temperature both in the solid state and in toluene solution to give the free phosphine-borane (Me3Si)2{Me2(H3B)P}CH (5) as the sole phosphorus-containing product. In addition, compounds 3, 4, and 4a react rapidly with THF in toluene solution, yielding 5 as the sole phosphorus-containing product; in contrast, compounds 7 and 8 are stable toward this solvent. PMID- 17425306 TI - Protein binding to lanthanide(III) complexes can reduce the water exchange rate at the lanthanide. AB - The GdIII-based magnetic resonance imaging contrast agent MS-325 targets the blood protein serum albumin, resulting in an increased efficacy (relaxivity) as a relaxation agent. MS-325 showed different relaxivities when bound to serum albumin from different species, e.g., r1=30.5 mM-1 s-1 (rabbit) vs 46.3 mM-1 s-1 (human) at 35 degrees C and 0.47 T. To investigate the mechanism for this difference, surrogate complexes were prepared where the GdIII ion was replaced by other LnIII ions. Fluorescence lifetime measurements of the EuIII analogue indicated that the hydration number was q=1 and did not change when bound to either human, rat, rabbit, pig, or dog serum albumin. The YbIII analogue, YbL1, was prepared and characterized by 1H NMR. Line-shape analysis of the paramagnetic shifted 1H NMR resonances in the presence of increasing amounts of human (HSA) or rabbit (RSA) serum albumin allowed estimation of the transverse relaxation rate, R2, of these resonances for the protein-bound YbL1. The rotational correlation time of YbL1 was calculated from R2, and the Yb-H distance and was tauR=8+/-1 ns when bound to HSA and 13+/-2 ns when bound to RSA. The water exchange rate at the DyIII analogue, DyL1, was determined from variable-temperature R2 measurements at 9.4 T when DyL1 was bound to either HSA or RSA. At 37 degrees C, water exchange at DyL1 was (31+/-5)x10(6) s-1 when bound to HSA but (3.8+/-0.2)x10(6) s-1 when bound to RSA. Slower water exchange upon RSA binding explains the differences in relaxivity observed. The approach of using surrogate lanthanides to identify specific molecular parameters influencing relaxivity is applicable to other protein-targeted GdIII contrast agents. PMID- 17425307 TI - Oxidation of the tris(carbene)borate complex PhB(MeIm)3MnI(CO)3 to MnIV[PhB(MeIm)3]2(OTf)2. AB - Reaction of the tris(carbene)borate ligand PhB(MeIm)3- with [Mn(CO)3(tBuCN)Br]2 leads to the manganese(I) tricarbonyl complex PhB(MeIm)3Mn(CO)3. In contrast to related complexes that are air-stable, PhB(MeIm)3Mn(CO)3 is O2-sensitive and is converted to a homoleptic MnIV complex. IR and cyclic voltammetry measurements of these complexes establish the exceptionally strong donating nature of the tris(carbene)borate ligand. PMID- 17425308 TI - DNA alkylation by pyrrole-imidazole seco-CBI conjugates with an indole linker: sequence-specific DNA alkylation with 10-base-pair recognition through heterodimer formation. AB - The sequence-specific DNA alkylation by conjugates 4 and 5, which consist of N methylpyrrole (Py)-N-methylimidazole (Im) polyamides and 1-(chloromethyl)-5 hydroxy-1,2-dihydro-3H-benz[e]indole (seco-CBI) linked with an indole linker, was investigated in the absence or presence of partner Py-Im polyamide 6. High resolution denaturing polyacrylamide gel electrophoresis revealed that conjugate 4 alkylates DNA at the sequences 5'-(A/T)GCCTA-3' through hairpin formation, and alkylates 5'-GGAAAGAAAA-3' through an extended binding mode. However, in the presence of partner Py-Im polyamide 6, conjugate 4 alkylates DNA at a completely different sequence, 5'-AGGTTGTCCA-3'. Alkylation of 4 in the presence of 6 was effectively inhibited by a competitor 7. Surface plasmon resonance (SPR) results indicated that conjugate 4 does not bind to 5'-AGGTTGTCCA-3', whereas 6 binds tightly to this sequence. The results suggest that alkylation proceeds through heterodimer formation, indicating that this is a general way to expand the recognition sequence for DNA alkylation by Py-Im seco-CBI conjugates. PMID- 17425309 TI - Hierarchical assembly of model cell surfaces: synthesis of mucin mimetic polymers and their display on supported bilayers. AB - Molecular level analysis of cell-surface phenomena could benefit from model systems comprising structurally defined components. Here we present the first step toward bottom-up assembly of model cell surfaces-the synthesis of mucin mimetics and their incorporation into artificial membranes. Natural mucins are densely glycosylated O-linked glycoproteins that serve numerous functions on cell surfaces. Their large size and extensive glycosylation makes the synthesis of these biopolymers impractical. We designed synthetically tractable glycosylated polymers that possess rodlike extended conformations similar to natural mucins. The glycosylated polymers were end-functionalized with lipid groups and embedded into supported lipid bilayers where they interact with protein receptors in a structure-dependent manner. Furthermore, their dynamic behavior in synthetic membranes mirrored that of natural biomolecules. This system provides a unique framework with which to study the behavior of mucin-like macromolecules in a controlled, cell surface-mimetic environment. PMID- 17425310 TI - Design and synthesis of a library of BODIPY-based environmental polarity sensors utilizing photoinduced electron-transfer-controlled fluorescence ON/OFF switching. AB - We systematically examined the mechanism of the solvent polarity dependence of the fluorescence ON/OFF threshold of the BODIPY (boron dipyrromethene) fluorophore and the role of photoinduced electron transfer (PeT). In a series of BODIPY derivatives with variously substituted benzene moieties at the 8-position, the oxidation potential of the benzene moiety became more positive and the reduction potential of the BODIPY fluorophore became more negative as the solvent polarity was decreased; consequently, the free energy change of PeT from the benzene moiety becomes larger in a more nonpolar environment. Utilizing this finding, we designed and synthesized a library of probes in which the threshold of fluorescence ON/OFF switching corresponds to different levels of solvent polarity. These environment-sensitive probes were used to examine bovine serum albumin (BSA) and living cells. The polarity at the surface of albumin was concluded to be similar to that of acetone, while the polarity of the internal membranes of HeLa cells was similar to that of dichloromethane. PMID- 17425311 TI - Direct measurement by laser flash photolysis of intraprotein electron transfer in a rat neuronal nitric oxide synthase. AB - Intraprotein interdomain electron transfer (IET) from flavin mononucleotide (FMN) to heme is essential in nitric oxide (NO) synthesis by NO synthase (NOS). Our previous laser flash photolysis studies have provided a direct determination of the kinetics of IET between the FMN and heme domains in truncated oxyFMN constructs of rat neuronal NOS (nNOS) and murine inducible NOS (iNOS), in which only the oxygenase and FMN domains along with the calmodulin (CaM) binding site are present [Feng, C. J.; Tollin, G.; Holliday, M. A.; Thomas, C.; Salerno, J. C.; Enemark, J. H.; Ghosh, D. K. Biochemistry 2006, 45, 6354-6362. Feng, C. J.; Thomas, C.; Holliday, M. A.; Tollin, G.; Salerno, J. C.; Ghosh, D. K.; Enemark, J. H. J. Am. Chem. Soc. 2006, 128, 3808-3811]. Here, we report the kinetics of IET between the FMN and heme domains in a rat nNOS holoenzyme in the presence and absence of added CaM using laser flash photolysis of CO dissociation in comparative studies on partially reduced NOS and a single domain NOS oxygenase construct. The IET rate constant in the presence of CaM is 36 s-1, whereas no IET was observed in the absence of CaM. The kinetics reported here are about an order of magnitude slower than the kinetics in a rat nNOS oxyFMN construct with added CaM (262 s-1). We attribute the slower IET between FMN and heme in the holoenzyme to the additional step of dissociation of the FMN domain from the reductase complex before reassociation with the oxygenase domain to form the electron transfer competent output state complex. This work provides the first direct measurement of CaM-controlled electron transfer between catalytically significant redox couples of FMN and heme in a nNOS holoenzyme. PMID- 17425313 TI - Hydrogen storage in a prototypical zeolitic imidazolate framework-8. PMID- 17425312 TI - Induced fit conformational changes of a "reversed amidine" heterocycle: optimized interactions in a DNA minor groove complex. AB - To better understand the molecular basis for recognition of the DNA minor groove by heterocyclic cations, a series of "reversed amidine" substituted heterocycles has been prepared. Amidine derivatives for targeting the minor groove have the amidine carbon linked to a central heterocyclic system, whereas in the reverse orientation, an amidine nitrogen provides the link. The reverse system has a larger dihedral angle as well as a modified spatial relationship with the groove relative to amidines. Because of the large dihedral, the reversed amidines should have reduced binding to DNA relative to similar amidines. Such a reduction is observed in footprinting, circular dichroism (CD), biosensor-surface plasmon resonance (SPR), and isothermal titration calorimetric (ITC) experiments with DB613, which has a central phenyl-furan-phenyl heterocyclic system. The reduction is not seen when a pyrrole (DB884) is substituted for the furan. Analysis of a number of derivatives defines the pyrrole and a terminal phenyl substituent on the reversed amidine groups as critical components in the strong binding of DB884. ITC and SPR comparisons showed that the better binding of DB884 was due to a more favorable binding enthalpy and that it had exceptionally slow dissociation from DNA. Crystallographic analysis of DB884 bound to an AATT site shows that the compound was bound in the minor groove in a 1:1 complex as suggested by CD solution studies. Surprisingly, unlike the amidine derivative, the pyrrole -NH of DB884 formed an H-bond with a central T of the AATT site and this accounts for the enthalpy-driven strong binding. The structural results and molecular modeling studies provide an explanation for the differences in binding affinities for related amidine and reversed amidine analogues. PMID- 17425314 TI - Design of chiral auxiliaries for the allene ether nazarov cyclization. PMID- 17425315 TI - Photochemical gating of intracellular Ca2+ release channels. PMID- 17425316 TI - Direct measurements of deuterium kinetic isotope effects in anionic, gas-phase substitution and elimination reactions. PMID- 17425317 TI - Chemical-shift anisotropy measurements of amide and carbonyl resonances in a microcrystalline protein with slow magic-angle spinning NMR spectroscopy. PMID- 17425318 TI - Biophysical characterization of a beta-peptide bundle: comparison to natural proteins. PMID- 17425319 TI - One-Step Synthesis and Magnetic Phase Transformation of Ln-TM-B Alloy by Chemical Reduction. AB - Binary and ternary intermetallic alloy systems are of interest for a variety of academic and technological applications. Despite recent advances in synthesizing binary alloy, there are very few reports of ternary alloy related to lanthanide series. The purpose of this work is to contribute to ternary alloy systems such as lanthanide-transition metal-boron with a simple chemical method and analysis of its magnetic behavior. Ternary Nd-Fe-B amorphous alloy was successfully synthesized with borohydride. The magnetic behavior in the process of formation of ternary Nd-Fe-B alloy and Nd2Fe14B from amorphous phase alloy is reported. Compared with the synthesis of a transition metal, the existence of a lanthanide ion makes aggregates-like particles with a diameter of 2 nm possible in the formation of a nanosphere, which is a significantly important result in terms of acceleration of the reduction-diffusion reaction for the formation of ternary alloy. In the process of reduction and diffusion, the Nd phase is diffused into the Fe-based phase, and then the ternary Nd2Fe14B intermetallic compound is fabricated. PMID- 17425320 TI - Versatile solid phase synthesis of gold nanoparticle dimers using an asymmetric functionalization approach. PMID- 17425321 TI - Chiral calcium complexes as Bronsted base catalysts for asymmetric addition of alpha-amino acid derivatives to alpha,beta-unsaturated carbonyl compounds. PMID- 17425322 TI - Design of C2-symmetric tetrahydropentalenes as new chiral diene ligands for highly enantioselective Rh-catalyzed arylation of N-tosylarylimines with arylboronic acids. PMID- 17425323 TI - Azide-modified graphitic surfaces for covalent attachment of alkyne-terminated molecules by "click" chemistry. PMID- 17425326 TI - Synthesis of a fluorous ligand and its application for asymmetric addition of dimethylzinc to aldehydes. AB - A new fluorous ligand was synthesized from the acetonide of dimethyl tartarate, which showed excellent asymmetric induction on the addition of dimethylzinc to aldehydes. This ligand will be useful for synthesis of bioactive compounds with a methyl carbinol moiety. It could be recycled without using a fluorous solvent or a fluorous column. PMID- 17425327 TI - Highly diastereoselective synthesis of vicinal quaternary and tertiary stereocenters using the iodo-aldol cyclization. AB - The intramolecular iodo-aldol cyclization of alpha-substituted enoate aldehydes and ketones is described. Using prochiral starting materials, the reaction produces hetero- and carbocycles containing quaternary centers adjacent to secondary or tertiary centers. The reactions occur in good yields and are highly selective for the trans-products, having the hydroxyl and iodomethyl groups on opposite faces of the ring system. PMID- 17425328 TI - Agrobacterium-mediated viral vector-amplified transient gene expression in Nicotiana glutinosa plant tissue culture. AB - A viral vector based on the bean yellow dwarf virus was investigated for its potential to increase transient gene expression. An intron-containing GUS reporter gene and the cis-acting viral regulatory elements were incorporated in the viral vector and could be complemented by the viral replication-associated proteins provided on a secondary vector. All vectors were delivered to Nicotiana glutinosa plant cell suspension or hairy root cultures via auxotrophic Agrobacterium tumefaciens. Cell culture generated greater yield of reporter gene expression than did root culture, as a result of the limitation imposed on roots to express the protein only in surface tissue containing actively dividing cells. Reporter gene expression increased for cell culture when the reporter gene construct was co-delivered with the construct supplying both viral replication associated proteins (REP and REPA); gene expression decreased when the construct supplying only the viral REP protein was co-delivered. Reporter protein expression increased from 0.091% for the reporter construct alone to 0.22% total soluble protein (% TSP) when the viral Rep-supplying vector was co-delivered with the reporter gene construct. Reporter protein was generated 3 days after the initiation of bacterial co-culture, providing for rapid generation of heterologous protein in cell culture. PMID- 17425329 TI - Protein misfolding and aggregation. AB - Interest in the problem of protein misfolding and aggregation has exploded in recent years for two reasons: (1) the sharp rise in the number and volume of therapeutic proteins produced commercially and (2) the recognition of the central role of protein aggregates in degenerative diseases. The systematic study of protein aggregation presents major challenges to both the experimentalist and the theoretician. Much of the work retains an empirical flavor due to the experimental complexities; the sensitivity of protein aggregation to the slightest change in protein amino acid composition, solvent properties, or protein concentration; and the lack of robust theoretical models of misfolding and aggregation. Novel experimental and computational approaches are being developed, and we anticipate substantial progress will be made in the near future. Several presentations describing the latest advances in protein misfolding and aggregation were given at the American Chemical Society meeting (BIOT division) held in September, 2006 in San Francisco. PMID- 17425330 TI - The linker between SH2 and kinase domains positively regulates catalysis of the Tec family kinases. AB - Tec family nonreceptor tyrosine kinases are key immunological enzymes that control processes that range from T and B cell development to reorganization of the actin cytoskeleton. The full-length Tec kinases have been resistant to crystallization. This lack of structural data and the paucity of in vitro biochemical data for this kinase family leave a void in our understanding of Tec kinase regulation. In this report we have used interleukin-2 tyrosine kinase (Itk) as a model system to gain insight into the regulatory apparatus of the Tec kinases. Use of a quantitative in vitro kinase assay has uncovered an essential role for the short linker region flanked by the SH2 and kinase domains of Itk in positively regulating Itk catalytic activity. The precise residues that allosterically regulate Itk are conserved among Tec kinases, pointing to the conserved nature of this regulatory mechanism within the family. These findings indicate that Tec kinases are not regulated in the same manner as the Src kinases but rather share some of the regulatory features of Csk instead. PMID- 17425331 TI - The unfolding pathway for Apo Escherichia coli aspartate aminotransferase is dependent on the choice of denaturant. AB - The guanidine hydrochloride (GdnHCl) mediated denaturation pathway for the apo form of homodimeric Escherichia coli aspartate aminotransferase (eAATase) (molecular mass = 43.5 kDa/monomer) includes a partially folded monomeric intermediate, M* [Herold, M., and Kirschner, K. (1990) Biochemistry 29, 1907 1913; Birolo, L., Dal Piaz, F., Pucci, P., and Marino, G. (2002) J. Biol. Chem. 277, 17428-17437]. The present investigation of the urea-mediated denaturation of eAATase finds no evidence for an M* species but uncovers a partially denatured dimeric form, D*, that is unpopulated in GdnHCl. Thus, the unfolding process is a function of the employed denaturant. D* retains less than 50% of the native secondary structure (circular dichroism), conserves significant quaternary and tertiary interactions, and unfolds cooperatively (mD*<==>U = 3.4 +/- 0.3 kcal mol 1 M-1). Therefore, the following equilibria obtain in the denaturation of apo eAATase: D <==> 2M 2M* <==> 2U in GdnHCl and D <==> D* <==> 2U in urea (D = native dimer, M = folded monomer, and U = unfolded state). The free energy of unfolding of apo-eAATase (D <==> 2U) is 36 +/- 3 kcal mol-1, while that for the D* 2U transition is 24 +/- 2 kcal mol-1, both at 1 M standard state and pH 7.5. PMID- 17425332 TI - Effect of zinc, copper, and calcium on the structure and stability of serum amyloid A. AB - Serum amyloid A (SAA) is a highly conserved acute phase reactant protein, and its concentration in serum can increase up to approximately 1000 times after an inflammatory stimuli. SAA is mainly associated with high-density lipoproteins in serum, and its main function appears to involve cholesterol transport and lipid metabolism. However, SAA has also been associated with many other functions and a number of diseases, although these potential links remain poorly understood. The three-dimensional structure of SAA is not known, but we have shown that murine SAA2.2 can exist in solution as a marginally stable hexamer, which at 37 degrees C dissociates to a monomeric species that misfolds irreversibly and self assembles into amyloid fibrils. Thus, the structure and function of SAA in vivo appear to be modulated when it binds to other proteins or small ligands. Herein, the effect of copper (Cu2+), zinc (Zn2+), and calcium (Ca2+) on the structure and stability of SAA2.2 in aqueous solution was examined using various probes of quaternary, tertiary, and secondary structure. At different concentrations of metals, including those found in the serum, the results show that the structure and stability of SAA2.2 are differently affected depending on the metal type and concentration. Copper (10-100 microM) was found to shift the equilibrium from hexamer to monomer without affecting significantly the stability of the tertiary and secondary structure of SAA2.2. In contrast, zinc (1-10 microM) bound to SAA2.2 and stabilized its quaternary, tertiary, and secondary structure. Calcium (1-10 mM) destabilized all elements of SAA2.2 structure and induced its aggregation at 10 mM. Complete aggregation of SAA2.2 was also observed when it was incubated with 1 mM Cu2+ or Zn2+, further demonstrating the tenuous structure and stability of SAA2.2. Thus, these results suggest that the many functional and pathological roles attributed to SAA may rely on its precarious structure, modulated by its interaction with ligands under homeostasis conditions and during the acute phase response. PMID- 17425333 TI - Thermodynamics of the interaction between O-acetylserine sulfhydrylase and the C terminus of serine acetyltransferase. AB - Cysteine biosynthesis in plants is partly regulated by the physical association of O-acetylserine sulfhydrylase (OASS) and serine acetyltransferase (SAT). Interaction of OASS and SAT requires only the 10 C-terminal residues of SAT. Here we analyze the thermodynamics of formation of a complex of Arabidopsis thaliana OASS (AtOASS) and the C-terminal ligand of AtSAT (C10 peptide) as a function of temperature and salt concentration using fluorescence spectroscopy and isothermal titration calorimetry (ITC). Our results suggest that the C-terminus of AtSAT provides the major contribution to the total binding energy in the plant cysteine synthase complex. The C10 peptide binds to the AtOASS homodimer in a 2:1 complex. Interaction between AtOASS and the C10 peptide is tight (Kd = 5-100 nM) over a range of temperatures (10-35 degrees C) and NaCl concentrations (0.02-1.3 M). AtOASS binding of the C10 peptide displays negative cooperativity at higher temperatures. ITC studies reveal compensating changes in the enthalpy and entropy of binding that also depend on temperature. The enthalpy of interaction has a significant temperature dependence (DeltaCp = -401 cal mol-1 K-1). The heat capacity change and salt dependence studies suggest that hydrophobic interactions drive formation of the AtOASS.C10 peptide complex. The potential regulatory effect of temperature on the plant cysteine synthase complex is discussed. PMID- 17425334 TI - Interaction of the fibronectin COOH-terminal Fib-2 regions with fibrin: further characterization and localization of the Fib-2-binding sites. AB - Incorporation of fibronectin into fibrin clots is important for the formation of a provisional matrix that promotes cell adhesion and migration during wound healing. Previous studies revealed that this incorporation occurs through noncovalent interaction between two NH2-terminal Fib-1 regions of fibronectin (one on each chain) and the alphaC-regions of fibrin, and is further reinforced by factor XIIIa-mediated covalent cross-linking of fibronectin to the fibrin matrix. To clarify the role of another pair of fibrin-binding regions, Fib-2, located at the disulfide-linked COOH-terminal ends of fibronectin, we prepared by limited proteolysis a dimeric 140 kDa (Fib-2)2 fragment containing both Fib-2 regions and tested its interaction with recombinant fragments corresponding to the alphaC regions of fibrin(ogen). In both ELISA and surface plasmon resonance (SPR) experiments 140 kDa (Fib-2)2 bound to the immobilized Aalpha221-610 alphaC fragment. However, the affinity of binding was substantially lower than that for Fib-1. Ligand blotting and ELISA established that the Fib-2 binding site is located in the connector part of the alphaC region including residues Aalpha221 391. Analysis of the SPR-detected binding of fibronectin to the immobilized Aalpha221-610 alphaC-fragment revealed two types of fibronectin-binding sites, one with high affinity and another one with much lower affinity. Competition experiments revealed about 30% inhibition of the Fib-2 mediated binding by increasing concentrations of Fib-1 fragment suggesting partial overlap of the two sets of binding sites. Based on these results and our previous studies we propose a mechanism of interaction of fibronectin with fibrin in which both Fib-1 and Fib 2 play a role. PMID- 17425335 TI - Improved chemical synthesis and demonstration of the relaxin receptor binding affinity and biological activity of mouse relaxin. AB - The primary stored and circulating form of relaxin in humans, human gene-2 (H2) relaxin, has potent antifibrotic properties with rapidly occurring efficacy. However, when administered to experimental models of fibrosis, H2 relaxin can only be applied over short-term (2-4 week) periods, due to rodents mounting an antibody response to the exogenous human relaxin, resulting in delayed clearance and, hence, increased and variable circulating levels. To overcome this problem, the current study investigated the therapeutic potential of mouse relaxin over long-term exposure in vivo. Mouse relaxin is unique among the known relaxins in that it possesses an extra residue within the C-terminal region of its A-chain. To enable a detailed assessment of its receptor interaction and biological properties, it was chemically synthesized in good overall yield by the separate preparation of each of its A- and B-chains followed by regioselective formation of each of the intramolecular and two intermolecular disulfide bonds. Murine relaxin was shown to bind with high affinity to the human, mouse, and rat RXFP1 (primary relaxin) receptor but with a slightly lower affinity to that of H2 relaxin. When administered to relaxin-deficient mice (which undergo an age dependent progression of organ fibrosis) over a 4 month treatment period, mouse relaxin was able to significantly inhibit the progression of collagen accumulation in several organs including the lung, kidney, testis, and skin (all p < 0.05 vs untreated group), consistent with the actions of H2 relaxin. These combined data demonstrate that mouse relaxin can effectively inhibit collagen deposition and accumulation (fibrosis) over long-term treatment periods. PMID- 17425336 TI - Coumarin-caged glycine that can be photolyzed within 3 microseconds by visible light. AB - The synthesis and characterization of a new photolabile precursor of glycine (coumarin-caged glycine) are reported. The new compound is suitable for rapid chemical kinetic investigations of the membrane-bound neurotransmitter receptor activated by glycine. Unlike previously used caging groups for glycine, this precursor can be photolyzed rapidly and efficiently in the visible wavelength region. This allows the use of a relatively inexpensive light source. The alpha carboxyl group of glycine was covalently coupled to the 7-(diethylamino)coumarin (DECM) caging group. The caged compound has a major absorption band with a maximum at 390 nm (epsilon390 = 13,900 M-1 cm-1). Photolysis was performed at wavelengths of >or=400 nm (epsilon400 = 12,400 M-1 cm-1). Under physiological conditions, DECM-caged glycine is water soluble and stable. In the visible wavelength region, it photolyzes rapidly to release glycine with a half-life of approximately 2.5 micrometers and a quantum yield of 0.12 +/- 0.01. The experimental results demonstrated that neither DECM-caged glycine nor its byproduct inhibits or activates human alpha1 glycine receptors expressed on the surface of HEK 293 cells. PMID- 17425337 TI - Inhibition of VMAT-2 and DT-diaphorase induce cell death in a substantia nigra derived cell line--an experimental cell model for dopamine toxicity studies. AB - We have induced intracellular dopamine oxidation to aminochrome in RCSN-3 cells derived from rat substantia nigra by inhibiting VMAT-2 with reserpine to increase free cytosolic dopamine concentration, to study aminochrome-dependent neurotoxicity in the absence of exogenous oxidizing agents such as metals, which may potentiate an aminochrome cytotoxic effect. The expression of VMAT-2 in RCSN 3 cells was determined by reverse transcriptase-polymerase chain reaction and immunocytochemistry. We observed double membrane bodies containing melanin when RCSN-3 cells were incubated with 100 microM dopamine by using transmission electron microscopy. No significant difference in the cell death was observed when the cells were treated 100 microM dopamine and 25 microM reserpine in the absence or presence of 100 microM dicoumarol, an inhibitor of DT-diaphorase. The lack of effect was due to the inhibitory action of 25 microM reserpine on DT diaphorase (Ki = 24 microM). However, a significant increase in the cell death was observed when DT-diaphorase was inhibited when the cells were incubated with 1 microM reserpine and 100 microM dopamine for 12 h since at this concentration reserpine inhibits VMAT-2 but not DT-diaphorase. Under this condition, we observed (i) the formation of blebbing; (ii) chromatin condensation accompanied by the formation of massive patches in contact with the nuclear membrane; (iii) the smoothness of the cell's surface, that is, lack of surface microprojections; and (iv) mitochondrial damage characterized by disruption of cristae architecture, which remains closely packed; disorganization of the mitochondrial matrix due to separation of the outer membrane from the internal membrane and considerable enlargement of the intermembrane space; and disruption of the external mitochondrial membrane determined by transmission electron microscopy. These results support the proposed neuroprotective role of DT-diaphorase against aminochrome neurotoxicity, and it suggests that RCSN-3 cells incubated with reserpine and dopamine are an excellent and more physiological cellular experimental model to study the role of dopamine oxidation in neurotoxic effects of dopamine. PMID- 17425338 TI - Retention of enzymatic activity of alpha-amylase in the reductive synthesis of gold nanoparticles. AB - In this paper, we report the generation of Au nanoparticles (NPs), using a pure enzyme for the reduction of AuCl4(-), with the retention of enzymatic activity in the complex. As a model system, alpha-amylase was used to readily synthesize and stabilize Au NPs in aqueous solution. Although several other enzymes were also pursued for the synthesis, it was interesting to observe that only alpha-amylase and EcoRI could produce Au NPs. Following NP synthesis, the activity of the enzyme was retained in the Au NP-alpha-amylase complex. The presence of Au NPs and alpha-amylase in the complex was established by UV-visible and FT-IR spectroscopy, X-ray diffraction (XRD) and transmission electron microscopic (TEM) measurements. Our observations suggest that the presence of free and exposed S-H groups is essential in the reduction of AuCl4(-) to Au NPs. Structural analysis of the enzymes showed that both alpha-amylase and EcoRI enzymes have free and exposed S-H groups in their native form and thus are suitable for the generation of NPs, whereas the other ones used here do not have such groups. Fortuitously, the enzymatic functional group of alpha-amylase is positioned opposite to that of the free and exposed S-H group, which makes it ideal for the production of Au NPs; binding of the enzyme to Au NPs via Au-S bond and also retention of the biological activity of the enzyme. PMID- 17425339 TI - NMR and IR study of fluorobenzene and hexafluorobenzene adsorbed on alumina. AB - The adsorption of fluorobenzene (C6H5F) and hexafluorobenzene (C6F6) onto the surface of neutral alumina is investigated by reflectance IR spectroscopy, near IR spectroscopy, and measurement of 19F NMR chemical shift values. Chemical shifts are dependent on surface coverage and reveal multiple peaks where different adsorption environments occur. C6H5F appears to be adsorbed through polar interactions with surface hydroxyls, whereas C6F6 shows separate resonances for the first layer of coverage and outer layers. Available surface areas are estimated, and reorientation of adsorbed hexafluorobenzene is proposed to account for chemical shift behavior and differences between calculated and measured monolayer coverage. PMID- 17425340 TI - Multilevel surface nano- and microstructuring via sequential photoswelling of dichromated gelatin. AB - Photoswelling of thin films of dichromated gelatin provides a basis for fabrication of multilevel surface reliefs via sequential UV illumination through different photomasks. The remarkable feature of this simple, benchtop technique is that by adjusting irradiation times, film thickness, or its hydration state the heights of the developed features can be varied from few nanometers to tens of microns. After UV exposure, the surface structures can be replicated faithfully into either soft or hard PDMS stamps. PMID- 17425341 TI - Difference in the bonding scheme of calix(6)arene and p-sulfonic calix(6)arene to nanoparticles of Fe(2)O(3) and Fe(3)O(4). AB - Calix(6)arene (C6) and p-sulfonic calix(6)arene (p-C6) formed chemical bonds with Fe(2)O(3) and Fe(3)O(4), respectively. The complexes exhibit different nature of bonding of C6 and p-C6 to the substrates. PMID- 17425342 TI - Synthesis and swelling behavior of pH-responsive polybase brushes. AB - We synthesize polybase brushes and investigate their swelling behavior. Poly(2 (dimethylamino)ethyl methacrylate)) (PDMAEMA) brushes are prepared by the "grafting from" method using surface-initiated Atom Transfer Radical Polymerization to obtain dense brushes with relatively monodisperse chains (PDI = 1.35). In situ quaternization reaction can be performed to obtain poly(2 (trimethylamino)ethyl methacrylate)) (PTMAEMA) brushes. We determine the swollen thickness of the brushes using ellipsometry and neutron reflectivity techniques. Brushes are submitted to different solvent conditions to be investigated as neutral brushes and weak and strong polyelectrolyte brushes. The swelling of the brushes is systematically compared to scaling models. It should be pointed out that the scaling analysis of different types of brushes (neutral polymer and weak and strong polyelectrolyte brushes) is performed with identical samples. The scaling behavior of the PDMAEMA brush in methanol and the PTMAEMA brush in water is in good agreement with the predicted scaling laws for a neutral polymer brush in a good solvent and a polyelectrolyte brush in the osmotic regime. The salt induced contraction of the quaternized brush is observed for high salt concentration, in agreement with the predicted transition between the regimes of the osmotic brush and the salted brush. From the crossover concentration, we calculate the effective charge ratio of the brush following the Manning counterion condensation. We also use PDMAEMA brushes as pH-responsive polybase brushes. The swelling behavior of the polybase brush is intermediate with respect to the behavior of the neutral polymer brush in a good solvent and the behavior of the quenched polyelectrolyte brush, as expected. The effective charge ratio of the PDMAEMA brush is determined as a function of pH using the scaling law of the polyelectrolyte brush in the osmotic regime. PMID- 17425343 TI - A new model of protein adsorption kinetics derived from simultaneous measurement of mass loading and changes in surface energy. AB - We describe a novel technology based on changes in the resonant frequency of an acoustically actuated surface and use it to measure temporal changes in the surface energy gamma (N m(-1)) of an elastomeric polymer membrane due to the adsorption of macromolecules from aqueous solution. The resonant elastomeric surface-tension (REST) sensor permits simultaneous determination of mass loading kinetics and gamma(t) for a given adsorption process, thereby providing a multivariable data set from which to build and test models of the kinetics of adsorption at solid-liquid interfaces. The technique is used to measure gamma(t) during the adsorption of either sodium dodecyl sulfate (SDS) or hen egg-white lysozyme (HEWL) onto an acrylic polymer membrane. The adsorption of SDS is reversible and is characterized by a decrease in gamma over a time period that coincides with that required for the mass loading of the membrane. For the adsorption of HEWL labeled with Alexa Fluor 532 dye, gamma continues to change long after the surface concentration of labeled HEWL, measured by using the elastomeric polymer membrane as an optical waveguide, reaches steady state. Gradual but significant changes in gamma(t) are observed as long as the concentration of protein in the bulk solution, c(b), remains nonzero. HEWL remains adsorbed to the membrane when c(b) = 0, but changes in gamma(t) are not observed under this condition, indicating that the interaction of bound protein molecules with those free in solution contribute to the prolonged change in the surface energy. This observation has been used to define a new model for the kinetics of globular protein adsorption to a solid-liquid interface that includes a mechanism by which the molecules in the bulk can facilitate the desorption of a sorbate molecule or change the energetic states of adsorbed molecules and, thus, the overall surface energy. The model is shown to capture the unique features of protein adsorption kinetics, including the relatively fast mass loading, the much more gradual change in surface energy that does not cease until the protein is removed from the bulk, the rapid desorption of an incubation-time-dependent fraction of bound protein when the protein is removed from the bulk, and the fixing of the residual surface concentration and surface energy at constant values once the removal of reversibly bound protein and free protein is complete. PMID- 17425344 TI - Controlled fabrication of nanopillar arrays as active substrates for surface enhanced Raman spectroscopy. AB - Highly ordered gold nanopillar arrays were fabricated using anodized aluminum oxide (AAO) templates. Nanopillars with a dimension of 110 +/- 15 nm in vertical height and 75 +/- 10 nm in base diameter were formed with a density of 150 microm(-2). The ordered nanopillar arrays give reproducible surface-enhanced Raman scattering (SERS) at a detection limit of 10(-8) M using thionine as probing molecules. The enhancement by the Au nanopillar arrays was comparable with or better than that of dispersed gold nanoparticle SERS substrates. This work demonstrates a new technique for producing highly ordered and reproducible SERS substrates potentially applicable for chemical and biological assay. PMID- 17425345 TI - Steric crowding effects on target detection in an affinity biosensor. AB - This work quantifies the impact of steric crowding on whole antibody (Ab) receptor immobilization and target Ab detection and also demonstrates how the versatile biotin/streptavidin receptor immobilization system must be tuned to optimize target detection in designing biosensors. Results are demonstrated on a label-free optical biosensor fabricated from n-type macroporous porous silicon (PSi) with approximately 88-107 nm diameter pores. We employ a sandwich assay scheme comprising a linking chemistry (biotin/streptavidin) to attach biotinylated anti-rabbit IgG (receptor) to detect rabbit IgG (target). A "bottom up" approach was taken to investigate each layer of the sandwich assay to optimize target binding. Steric crowding was observed to hinder subsequent layer binding for each layer in the sandwich (biotin, streptavidin, and receptor). Our results give definitive evidence that the onset of steric crowding within the biotin layer occurs at a surface coverage of 57%, which is much higher compared to that from published work on well-ordered self-assembled biotin monolayers on planar gold surfaces. This difference is attributed to the topographical heterogeneity of the PSi substrate. Streptavidin (SA) binding to surface-linked biotin was altered by preblocking the streptavidin binding sites with biotin. Through consistent trends in data, preblocking SA was shown to reduce steric crowding within the SA layer, which translated into increased receptor immobilization. The final detection range of rabbit IgG was 0.07-3 mg mL(-1) (0.4 17 ng mm(-2)), and binding specificity was demonstrated by employing an anti chicken IgG control receptor. This study underlines the importance of considering binding avidity and surface topography in optimizing chip-based biosensors. PMID- 17425346 TI - Formation of Pt-Ru nanoparticles in ethylene glycol solution: an in situ X-ray absorption spectroscopy study. AB - The chemical state and formation mechanism of Pt-Ru nanoparticles (NPs) synthesized by using ethylene glycol (EG) as a reducing agent and their stability have been examined by in situ X-ray absorption spectroscopy (XAS) at the Pt LIII and Ru K edges. It appears that the reduction of Pt(IV) and Ru(III) precursor salts by EG is not a straightforward reaction but involves different intermediate steps. The pH control of the reaction mixture containing Pt(IV) and Ru(III) precursor salts in EG to 11 led to the reduction of Pt(IV) to Pt(II) corresponding to [PtCl4](2-) whereas Ru(III)Cl3 is changed to the [Ru(OH)6](3-) species. Refluxing the mixture containing [PtCl4](2-) and [Ru(OH)6](3-) species at 160 degrees C for 0.5 h produces Pt-Ru NPs as indicated by the presence of Pt and Ru in the first coordination shell of the respective metals. No change in XAS structural parameters is found when the reaction time is further increased, indicating that the Pt-Ru NPs formed are extremely stable and less prone to aggregation. XAS structural parameters suggest a Pt-rich core and a Ru-rich shell structure for the final Pt-Ru NPs. Due to the inherent advantages of the EG reduction method, the atomic distribution and alloying extent of Pt and Ru in the Pt-Ru NPs synthesized by the EG method are higher than those of the Pt-Ru/C NPs synthesized by a modified Watanabe method. PMID- 17425347 TI - Chemical modifications of inert organic monolayers with oxygen plasma for biosensor applications. AB - In this paper we present a study of using oxygen plasma for chemically modifying inert hydrocarbon self-assembled monolayers of octadecyltrichlorosilane (OTS SAMs) and rendering active surfaces for protein immobilization. Detailed surface modification and protein immobilization were characterized by using ellipsometry, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared-attenuated total reflectance spectroscopy, and fluorescence microscopy. Our XPS results showed that the surface reaction between OTS-SAMs and oxygen plasma can generate new surface functional groups such as alcohol (C-O), aldehyde (C=O), and carboxylic acid (O-C=O), and their compositions can be controlled by using different treatment times and powers. A short treatment time ( approximately 1 s) and high power (10 W) can lead to a higher density of aldehyde groups, which can serve as linker groups for protein immobilization through the formation of Schiff bases with the amine groups of proteins. By using the fluorescence immunostaining method, we confirmed that human immunoglobulin (IgG) can be immobilized on a glass slide, only if the surface was decorated with OTS-SAMs and if the OTS-SAMs were pretreated with oxygen plasma. The protein immobilized on the oxygen-plasma treated surface can only be recognized by using a highly specific antibody, FITC anti-IgG, but not FITC-anti-biotin. PMID- 17425348 TI - Silver nanoparticles capped by oleylamine: formation, growth, and self organization. AB - Nearly monodisperse silver nanoparticles have been prepared in a simple oleylamine-liquid paraffin system. Intensive study has found that the formation process of silver nanoparticles could be divided into three stages: growth, incubation, and Ostwald ripening stages. Ultraviolet-visible spectroscopy, transmission electron microscopy (TEM), and high-resolution TEM have all demonstrated the occurrence of Ostwald ripening, which could result in better control over the size and size distribution of silver nanoparticles. SAXS (small angle X-ray scattering) results show that the as-obtained silver nanoparticles can self-assemble into ordered arrays. The possible reduction mechanism of silver ions by oleylamine is related to the Ag+-mediated conversion of primary amines to nitriles. PMID- 17425349 TI - Top-down approaches to the formation of silica nanoparticle patterns. AB - This article reports a simple, versatile approach to the fabrication of lithographically defined mesoscopic colloidal silica nanoparticle patterns over large areas using spin-coating, interferometric lithography, and reactive-ion etching. One-dimensional nanoparticle films (bands) and 2D discs, diamonds, and holes with sub-micrometer periodicity, high quality, and excellent uniformity were successfully fabricated over large areas. The well-defined shape and period of the patterned nanoparticle film were controlled in the interferometric lithography step, while the thickness of nanoparticle film was easily tuned in the spin-coating step. This approach can extend to other deposition methods such as convective self-assembly, electrostatic self-assembly, and other materials such as metallic and ferromagnetic nanoparticles. We have also been able to generate sparse, random, isolated particle patterns, using a combination of interferometric lithography and layer-by-layer deposition as an extension of this approach to another deposition method, and to generate disc nanoparticle patterns using colloidal lithography as an extension of this approach to another lithography technique. These patterned films will find important applications in the fields of material growth, biosensors, and catalysis, as well as serving as building blocks for further fabrication. PMID- 17425350 TI - iTRAQ-coupled 2D LC-MS/MS analysis on protein profile in vascular smooth muscle cells incubated with S- and R-enantiomers of propranolol: possible role of metabolic enzymes involved in cellular anabolism and antioxidant activity. AB - Propranolol is a nonselective beta-blocker of the beta-adrenergic receptors, and the S-enantiomer is more active compared with the R-enantiomer. Clinically, it has been shown to be effective in hypermetabolic burn patients by decreasing cardiac work, protein catabolism, and lipolysis. While gene expression profiles have recently been reported in children receiving propranolol treatment, variations from one individual to another may have influenced the data analysis. Using iTRAQ-coupled 2D LC-MS/MS analysis, we report here the first study of protein profile in vascular smooth muscle cells incubated separately with the two enantiomers of propranolol. Four types of cellular proteins including metabolic enzymes, signaling molecules, cytoskeletal proteins, and those involved in DNA synthesis/protein translation displayed changes. The higher protein level of a number of enzymes involved in cellular anabolism and antioxidant activity in cells incubated with the S-enantiomer, as revealed by LC-MS/MS, was further supported by real-time PCR and Western blot analyses. Significantly, the increase in the anabolic activity associated with the higher level of metabolic enzymes was also supported by the higher intracellular concentration of the metabolic cofactor NAD+ which was a result of an increased oxidation of NADH. Our findings therefore provide molecular evidence on metabolic effect associated with propranolol treatment. The metabolic enzymes identified in our study may in turn be useful targets for future pharmaceutical interventions to reduce clinical side effects following propranolol treatment. PMID- 17425351 TI - Tetraphenylporphyrin-cobalt(III) bis(1,2-dicarbollide) conjugates: from the solution characteristics to inhibition of HIV protease. AB - Tetraphenylporphyrin conjugates with one (PB1) and four (PB4) cobalt(III) bis(1,2 dicarbollide) substituents were synthesized and the physicochemical and photophysical properties as well as inhibition of HIV-1 protease were described. In methanol, both PB1 and PB4 were monomeric producing the triplet states and singlet oxygen after excitation. The triplet states of PB4 were quickly protonated. Porphyrins exhibited a small decrease of the quantum yields of the singlet oxygen formation (17% for PB4 and 13% for PB1) as compared with 5,10,15,20-tetrakis(4-sulfonatophenyl)porphyrin. On the contrary, no singlet oxygen was detected in aqueous solutions because of strong aggregation. Light scattering and atomic force microscopy (AFM) measurements documented that the behavior of aggregates in aqueous solutions is fairly complex and depends on pH, concentration, and aging. The aggregation started from spherical particles in neutral solutions. In acidic solutions, extended aggregation occurred because of slow protonation of the porphyrin pyrrole nitrogen atoms. Both PB1 and PB4 are new representatives of nonpeptide HIV-1 protease inhibitors. Their activity increased with the increasing number of the cobalt(III) bis(1,2-dicarbollide) substituents and was characterized with the IC50 values of 290+/-44 nM for PB1 and 77+/-13 nM for PB4. PMID- 17425352 TI - The mechanism of cyclic nucleotide hydrolysis in the phosphodiesterase catalytic site. AB - The cyclic nucleotide phosphodiesterase superfamily of enzymes (PDEs) catalyzes the stereospecific hydrolysis of the second messengers adenosine and guanosine 3',5'- cyclic monophosphate (cAMP, cGMP) to produce 5'-AMP and 5'-GMP, respectively. The PDEs are targets of high-throughput screening to determine selective inhibitors for a variety of therapeutic purposes. The catalytic pocket where the hydrolysis takes place is a highly conserved region and has several residues which are absolutely conserved across the PDE families. In this study, we consider a model cyclic substrate in which the adenine/guanine base has been replaced with a hydrogen atom, and we present results of a quantum computational investigation of the hydrolysis reaction as it occurs within the PDE catalytic site using the ONIOM hybrid (B3LYP/6-31g(d):PM3) method. We characterize the bound substrate, the bound hydrolyzed product, and the transition state which connects them for our model cyclic substrate placed in a truncated model of the PDE4D2 catalytic site. We address the role that the conserved histidine proximal to the bimetal system of the catalytic site, along with its conserved glutamine partner, plays in the generation of the hydroxide nucleophile. Our study provides computational evidence for several key features of the cAMP/cGMP hydrolysis mechanism as it occurs within the protein environment across the PDE superfamily. PMID- 17425353 TI - Electrochemistry and electrocatalysis of hemoglobin in Nafion/nano-CaCO3 film on a new ionic liquid BPPF6 modified carbon paste electrode. AB - Room temperature ionic liquid N-butylpyridinium hexafluorophosphate (BPPF6) was used as a binder to construct a new carbon ionic liquid electrode (CILE), which exhibited enhanced electrochemical behavior as compared with the traditional carbon paste electrode with paraffin. By using the CILE as the basal electrode, hemoglobin (Hb) was immobilized on the surface of the CILE with nano-CaCO3 and Nafion film step by step. The Hb molecule in the film kept its native structure and showed good electrochemical behavior. In pH 7.0 Britton-Robinson (B-R) buffer solution, a pair of well-defined, quasi-reversible cyclic voltammetric peaks appeared with cathodic and anodic peak potentials located at -0.444 and -0.285 V (vs SCE), respectively, and the formal potential (E degrees') was at -0.365 V, which was the characteristic of Hb Fe(III)/Fe(II) redox couples. The formal potential of Hb shifted linearly to the increase of buffer pH with a slope of 50.6 mV pH-1, indicating that one electron transferred was accompanied with one proton transportation. Ultraviolet-visible (UV-vis) and Fourier transform infrared (FT-IR) spectroscopy studies showed that Hb immobilized in the Nafion/nano-CaCO3 film still remained its native arrangement. The Hb modified electrode showed an excellent electrocatalytic behavior to the reduction of H2O2, trichloroacetic acid (TCA), and NaNO2. PMID- 17425354 TI - Lipid-based mechanisms for vesicle fission. AB - Shape transformations and topological changes of lipid vesicles, such as fusion, budding, and fission, have important chemical physical and biological significance. In this paper, we study the fission process of lipid vesicles. Two distinct routes are considered that are both based on an asymmetry of the lipid distribution within the membrane. This asymmetry consists of a nonuniform distribution of two types of lipids. In the first mechanism, the two types of lipids are equally distributed over both leaflets of the membrane. Phase separation of the lipids within both leaflets, however, results in the formation of rafts, which form buds that can split off. In the second mechanism, the asymmetry consists of a difference in composition between the two monolayers of the membrane. This difference in composition yields a spontaneous curvature, reshaping the vesicle into a dumbbell such that it can split. Both pathways are studied with molecular dynamics simulations using a coarse-grained lipid model. For each of the pathways, the conditions required to obtain complete fission are investigated, and it is shown that for the second pathway, much smaller differences between the lipids are needed to obtain fission than for the first pathway. Furthermore, the lipid composition of the resulting split vesicles is shown to be completely different for both pathways, and essential differences between the fission pathway and the pathway of the inverse process, i.e., fusion, are shown to exist. PMID- 17425355 TI - Two-dimensionally self-arranged protein nanoarrays on diblock copolymer templates. AB - Novel methods for creating protein arrays with two-dimensional control can significantly enhance basic biological research as well as various bioarray applications. We demonstrate that the structural variety and chemical heterogeneity of self-assembled, hexagonal polystyrene-b-poly(vinylpyridine) micelles can be successfully exploited as templates for easy and rapid fabrication of functional protein arrays over a large scale. Spontaneous formation of such polymeric template-guided protein molecules yields high-density protein arrays that exhibit repeat spacings in a nanoscopic dimension. The ensuing self-assembled protein molecules in the array maintain their natural conformation and activity over a very long time period. By tuning the size of the underlying block copolymer templates, our amphiphilic diblock copolymer-based approach to create high-density protein patterns also permits spatial control over two-dimensional repeat spacings of protein nanoarrays. These unique advantages of polystyrene-b-poly(vinylpyridine) templates make the spontaneously constructed protein nanoarrays highly suitable as functional protein sensor substrates. Therefore, our novel two-dimensional protein assembly method can be greatly beneficial for high-throughput proteomic assays and multiplexed high density protein sensing applications. PMID- 17425356 TI - Electron-transfer oxidation properties of substituted bi-, ter-, and quaterpyrroles. AB - A set of open-chain fully substituted bi-, ter-, and quaterpyrroles bearing analogous substituents in the alpha- and beta-pyrrolic positions were studied as a function of their chain length, subunit number, and size of potential conjugation pathway by means of cyclic voltammetry, EPR, and UV-vis spectroelectrochemistry. A comparison of E1/2 values for the first one-electron abstraction of bipyrrole 1 (1.07 V), terpyrrole 2 (0.67 V), and quaterpyrrole 3 (0.44 V) demonstrate clearly that the longer oligopyrroles are easier to oxidize. A strong absorption band is observed in the visible region when terpyrrole 2 is subject to one-electron oxidation, growing in at 856 nm accompanied by a shoulder at 778 nm. These strong absorptions in the visible region of the spectrum are in sharp contrast with the absence of bands in the red region when the bipyrrole 1 is subject to a similar one-electron oxidation and this can be explained by the presence of a longer conjugation pathway in the singly oxidized forms of 2 as was confirmed by EPR spectroscopy. The EPR spectra of 1*+, 2*+, and 3*+ indicate that the unpaired electron is more delocalized on the pyrroles with a longer conjugation and that the more the unpaired electron is delocalized, the faster is the electron exchange rate. PMID- 17425357 TI - Long-range Lennard-Jones and electrostatic interactions in interfaces: application of the isotropic periodic sum method. AB - Molecular dynamics (MD) simulations of heptane/vapor, hexadecane/vapor, water/vapor, hexadecane/water, and dipalmitoylphosphatidylcholine (DPPC) bilayers and monolayers are analyzed to determine the accuracy of treating long-range interactions in interfaces with the isotropic periodic sum (IPS) method. The method and cutoff (rc) dependences of surface tensions, density profiles, water dipole orientation, and electrostatic potential profiles are used as metrics. The water/vapor, heptane/vapor, and hexadecane/vapor interfaces are accurately and efficiently calculated with 2D IPS (rc=10 A). It is demonstrated that 3D IPS is not practical for any of the interfacial systems studied. However, the hybrid method PME/IPS [Particle Mesh Ewald for electrostatics and 3D IPS for Lennard Jones (LJ) interactions] provides an efficient way to include both types of long range forces in simulations of large liquid/vacuum and all liquid/liquid interfaces, including lipid monolayers and bilayers. A previously published pressure-based long-range LJ correction yields results similar to those of PME/IPS for liquid/liquid interfaces. The contributions to surface tension of LJ terms arising from interactions beyond 10 A range from 13 dyn/cm for the hexadecane/vapor interface to approximately 3 dyn/cm for hexadecane/water and DPPC bilayers and monolayers. Surface tensions of alkane/vapor, hexadecane/water, and DPPC monolayers based on the CHARMM lipid force fields agree very well with experiment, whereas surface tensions of the TIP3P and TIP4P-Ew water models underestimate experiment by 16 and 11 dyn/cm, respectively. Dipole potential drops (DeltaPsi) are less sensitive to long-range LJ interactions than surface tensions. However, DeltaPsi for the DPPC bilayer (845+/-3 mV proceeding from water to lipid) and water (547+/-2 mV for TIP4P-Ew and 521+/-3 mV for TIP3P) overestimate experiment by factors of 3 and 5, respectively, and represent expected deficiencies in nonpolarizable force fields. PMID- 17425358 TI - Advantages and limitations of the use of an extended polyelectrolyte model to describe the proton-binding process in macromolecular systems. Application to a poly(acrylic acid) and a humic acid. AB - A number of studies have shown the suitability of the polyelectrolyte model to describe the proton-binding behavior of macromolecules. This model, however, has two limitations associated with its theoretical approach: (1) it does not consider the possible heterogeneity of binding sites, and (2) for certain calculations, it involves the need to assume a specific molecular geometry. In this article we describe the theoretical basis of an extension of the polyelectrolyte model that removes the two limitations described above. Likewise, we discuss the advantages and limitations of the extended polyelectrolyte model (EPM) through its application to describe the proton-binding process in a well characterized macromolecular system (a poly(acrylic acid)) and a complex molecular system (a humic acid). The results obtained showed the suitability of EPM to describe proton-binding processes in complex molecular systems without the need to assume previously a specific molecular geometry and explicitly considering the possible heterogeneity of the binding sites. The results obtained indicated that the field effects associated with the conformational structure corresponding to each ionic strength, even in the discharged state, affect the values of the intrinsic constants defining the proton-binding process using EPM. Likewise, EPM analysis reveals the significant influence of both the surface charge density and the molecular size on the value of the electrostatic effects affecting the values of the intrinsic constants in the proton-binding process. PMID- 17425359 TI - Functional metalloproteins integrated with conductive substrates: detecting single molecules and sensing individual recognition events. AB - In the past decade, there has been significant interest in the integration of biomaterials with electronic elements: combining biological functions of biomolecules with nanotechnology offers new perspectives for implementation of ultrasensitive hybrid nanodevices. In particular, great attention has been devoted to redox metalloproteins, since they possess unique characteristics, such as electron-transfer capability, possibility of gating redox activity, and nanometric size, which make them appealing for bioelectronics applications at the nanoscale. The reliable connection of redox proteins to electrodes, aimed at ensuring good electrical contact with the conducting substrate besides preserving protein functionality, is a fundamental step for designing a hybrid nanodevice and calls for a full characterization of the immobilized proteins, possibly at the single-molecule level. Here, we describe how a multitechnique approach, based on several scanning probe microscopy techniques, may provide a comprehensive characterization of different metalloproteins on metal electrodes, disclosing unique information not only about morphological properties of the adsorbed molecules but also about the effectiveness of electrical coupling with the conductive substrate, or even concerning the preserved biorecognition capability upon adsorption. We also show how the success of an immobilization strategy, which is of primary importance for optimal integration of metalloproteins with a metal electrode, can be promptly assessed by means of the proposed approach. Besides the characterization aspect, the complementary employment of the proposed techniques deserves major potentialities for ultrasensitive detection of adsorbed biomolecules. In particular, it is shown how sensing of single metalloproteins may be optimized by monitoring the most appropriate observable. Additionally, we suggest how the combination of several experimental techniques might offer increased versatility, real-time response, and wide applicability as a detection method, once a reproducible correlation among signals coming from different single-molecule techniques is established. PMID- 17425360 TI - Interplay of electrostatic and hydrophobic effects with binding of cationic gemini surfactants and a conjugated polyanion: experimental and molecular modeling studies. AB - Understanding factors responsible for the fluorescence behavior of conjugated polyelectrolytes and modulation of their behavior are important for their application as functional materials. The interaction between the anionic poly{1,4 phenylene-[9,9-bis(4-phenoxy-butylsulfonate)]fluorene-2,7-diyl}copolymer (PBS PFP) and cationic gemini surfactants alpha,omega-(CmH2m+1N+(CH3)2)2(CH2)s(Br-)2 (m-s-m; m=12, s=2, 3, 5, 6, 10, and 12) has been studied experimentally in aqueous solution. These surfactants are chosen to see whether molecular recognition and self-assembly occurs between the oppositely charged conjugated polyelectrolyte and gemini surfactant when the spacer length on the surfactant is similar to the intercharge separation on the polymer. Without surfactants, PBS PFP exists as aggregates. These are broken up upon addition of gemini surfactants. However, as anticipated, the behavior strongly depends upon spacer length (s). Fluorescence measurements show three surfactant concentration regimes: At low concentrations (<2x10(-6) M) quenching occurs and is most marked with the small spacer 12-2-12; at intermediate concentrations (approximately 2x10(-6)-10(-3) M), fluorescence intensity is constant, with a 12-carbon spacer 12-12-12 showing the strongest fluorescence; above the critical micelle concentration (CMC; approximately 10(-3) M) increases in emission intensity are seen in all cases and are largest with the intermediate spacers 12-5-12 and 12-6 12, where the spacer length most closely matches the distance between monomer units on the polymer. With longer spacer length surfactants, surface tension measurements for concentrations below the CMC reveal the presence of polymer surfactant aggregates at the air-water interface, possibly reflecting increased hydrophobicity. Above the CMC, small-angle neutron scattering experiments for the 12-6-12 system show the presence of spherical aggregates, both for the pure surfactant and for polyelectrolyte/gemini mixtures. Molecular dynamics simulations help rationalize these observations and show that there is a very fine balance between electrostatic and hydrophobic interactions. With the shortest spacer 12-2-12, Coulombic interactions are dominant, while for the longest spacer 12-12-12 the driving force involves hydrophobic interactions. Qualitatively, with the intermediate 12-5-12 and 12-6-12 systems, the optimum balance is observed between Coulombic and hydrophobic interactions, explaining their strong fluorescence enhancement. PMID- 17425361 TI - Protons in non-ionic aqueous reverse micelles. AB - Using molecular dynamics techniques, we investigate the solvation of an excess proton within an aqueous reverse micelle in vacuo, with the neutral surfactant diethylene glycol monodecyl ether [CH3(CH2)11(OC2H4)2OH]. The simulation experiments were performed using a multistate empirical valence bond Hamiltonian model. Our results show that the stable solvation environments for the excess proton are located in the water-surfactant interface and that its first solvation shell is composed exclusively by water molecules. The relative prevalence of Eigen- versus Zundel-like solvation structures is investigated; compared to bulk results, Zundel-like structures in micelles become somewhat more stable. Characteristic times for the proton translocation jumps have been computed using population relaxation time correlation functions. The micellar rate for proton transfer is approximately 40x smaller than that found in bulk water at ambient conditions. Differences in the computed rates are examined in terms of the hydrogen-bond connectivity involving the first solvation shell of the excess charge with the rest of the micellar environment. Simulation results would indicate that proton transfers are correlated with rare episodes during which the HB connectivity between the first and second solvation shells suffers profound modifications. PMID- 17425362 TI - Free energy and dynamics of electron-transfer reactions in a room temperature ionic liquid. AB - Reaction free energetics and dynamics of unimolecular electron-transfer processes in ionic liquid 1-ethyl-3-methylimidazolium hexafluorophosphate (EMI+PF6-) are investigated via molecular dynamics computer simulations employing a model diatomic solute and compared with those in aprotic acetonitrile. Using the free energy perturbation method, diabatic free energy curves relevant to charge separation and recombination processes are studied over a wide range of the reaction coordinate. The diabatic curves are found to vary with the solute charge distribution, especially in EMI+PF6-. Nevertheless, if the free energy of reaction is not that substantial, the Marcus free energy relationship holds reasonably well, provided that the reorganization free energy averaged between the reactant and product states is employed. The effective polarity, measured as solvation-induced stabilization of dipolar solutes, is higher for EMI+PF6- than for acetonitrile, consonant with many solvatochromic measurements. Thus, in the normal regime, activation barriers for charge separation and recombination reactions are, respectively, lower and higher in EMI+PF6- than in acetonitrile. The influence of solvent dynamics on reaction kinetics through modulations of activation, deactivation, and barrier crossing is analyzed. Even though overall solvent relaxation dynamics in EMI+PF6- are considerably slower than those in acetonitrile, the deviation of the rate constant from the transition state theory predictions is found to be small for both solvents. Implications of this finding for other reactions in ionic liquids are briefly discussed. PMID- 17425363 TI - Poly(L-lysine)-graft-chitosan copolymers: synthesis, characterization, and gene transfection effect. AB - Polypeptide/polysaccharide graft copolymers poly(L-lysine)-graft-chitosan (PLL-g Chi) were prepared by ring-opening polymerization (ROP) of epsilon benzoxycarbonyl L-lysine N-carboxyanhydrides (Z-L-lysine NCA) in the presence of 6-O-triphenylmethyl chitosan. The PLL-g-Chi copolymers were thoroughly characterized by 1H NMR, 13C NMR, Fourier transform infrared (FT-IR), and gel permeation chromatography (GPC). The number-average degree of polymerization of PLL grafted onto the chitosan backbone could be adjusted by controlling the feed ratio of NCA to 6-O-triphenylmethyl chitosan. The particle size of the complexes formed from the copolymer and calf thymus DNA was measured by dynamic light scattering (DLS). It was found in the range of 120 approximately 340 nm. The gel retardation electrophoresis showed that the PLL-g-Chi copolymers possessed better plasmid DNA-binding ability than chitosan. The gene transfection effect in HEK 293T cells of the copolymers was evaluated, and the results showed that the gene transfection ability of the copolymer was better than that of chitosan and was dependent on the PLL grafting ratio. The PLL-g-Chi copolymers could be used as effective gene delivery vectors. PMID- 17425364 TI - Biodegradable poly(ester-anhydride) for new antifouling coating. PMID- 17425365 TI - The chemistry and applications of antimicrobial polymers: a state-of-the-art review. AB - Microbial infection remains one of the most serious complications in several areas, particularly in medical devices, drugs, health care and hygienic applications, water purification systems, hospital and dental surgery equipment, textiles, food packaging, and food storage. Antimicrobials gain interest from both academic research and industry due to their potential to provide quality and safety benefits to many materials. However, low molecular weight antimicrobial agents suffer from many disadvantages, such as toxicity to the environment and short-term antimicrobial ability. To overcome problems associated with the low molecular weight antimicrobial agents, antimicrobial functional groups can be introduced into polymer molecules. The use of antimicrobial polymers offers promise for enhancing the efficacy of some existing antimicrobial agents and minimizing the environmental problems accompanying conventional antimicrobial agents by reducing the residual toxicity of the agents, increasing their efficiency and selectivity, and prolonging the lifetime of the antimicrobial agents. Research concerning the development of antimicrobial polymers represents a great a challenge for both the academic world and industry. This article reviews the state of the art of antimicrobial polymers primarily since the last comprehensive review by one of the authors in 1996. In particular, it discusses the requirements of antimicrobial polymers, factors affecting the antimicrobial activities, methods of synthesizing antimicrobial polymers, major fields of applications, and future and perspectives in the field of antimicrobial polymers. PMID- 17425366 TI - Rapidly in situ-forming degradable hydrogels from dextran thiols through Michael addition. AB - Thiol-functionalized dextrans (dex-SH) (M(n,dextran) = 14K or 31K) with degrees of substitution (DS) ranging from 12 to 25 were synthesized and investigated for in situ hydrogel formation via Michael type addition using poly(ethylene glycol) tetra-acrylate (PEG-4-Acr) or a dextran vinyl sulfone conjugate with DS 10 (dex VS DS 10). Dex-SH was prepared by activation of the hydroxyl groups of dextran with 4-nitrophenyl chloroformate and subsequent reaction with cysteamine. Hydrogels were rapidly formed in situ under physiological conditions upon mixing aqueous solutions of dex-SH and either PEG-4-Acr or dex-VS DS 10 at polymer concentrations of 10 to 20 w/v%. Rheological studies showed that these hydrogels are highly elastic. By varying the DS, concentration, dextran molecular weight, and type of cross-linker, hydrogels with a broad range of storage moduli of 9 to 100 kPa could be obtained. Varying the ratio of thiol to vinyl sulfone groups from 0.9 to 1.1 did not alter the storage modulus of the hydrogels, whereas larger deviations from equimolarity (thiol to vinyl sulfone ratios of 0.75 and 1.5) considerably decreased the storage modulus. The plateau value of hydrogel storage modulus was reached much faster at pH 7.4 compared to pH 7, due to a higher concentration of the thiolate anion at higher pH. These hydrogels were degradable under physiological conditions. Degradation times were 3 to 7 weeks for dex-SH/dex-VS DS 10 hydrogels and 7 to over 21 weeks for dex-SH/PEG-4-Acr hydrogels, depending on the DS, concentration, and dextran molecular weight. PMID- 17425367 TI - Cholesterol mesogen containing water-soluble copolymers: design and organization behavior at different interfaces. AB - Cholesterol mesogen containing monomer, cholesteryl acrylamido butyrate (CAB) with the novel spacer group drawn from 4-amino butyric acid has been demonstrated to exhibit good reactivity with 2-acrylamido-2-methyl-1-propane sulfonic acid (AMPS) to yield copolymers with CAB content as high as 15 mol % hitherto not achieved. The spacer group is shown to provide the twin benefits of enhanced reactivity and solubility in water. The high pK(a) at > or =9.90 of these copolymers estimated from potentiometric studies demonstrates packing of AMPS segments as ionic clusters. The higher CAB in copolymer C provides the most densely packed nonpolar microdomains. From fluorescence quenching studies, the cross-linking provided by the cholesterol chains favoring intra- or intermolecular aggregated structures has been established. At the air/solution interface, copolymer C exhibits the most close-packed structures exhibiting "a" of 41.2 A(2)/molecule. The effect of neutralization on the adsorption characteristics is investigated. PMID- 17425368 TI - Independent generation of C5'-nucleosidyl radicals in thymidine and 2' deoxyguanosine. AB - The synthesis of the C5' tert-butyl ketone of thymidine 1a and 2'-deoxyguanosine 2 is achieved by reaction of 5'-C-cyano derivatives with tert-butyl lithium followed by acid hydrolysis. The 5'R configuration is assigned by X-ray crystal structure determination of an opportunely protected derivative of 1a. The (5'S) isomers of both nucleosides are not stable, and a complete decomposition occurs in the reaction medium. The photochemistry of 1a and 2 effectively produced the thymidin-5'-yl radical and the 2'-deoxyguanosin-5'-yl radical, respectively. In the thymidine system, the C5' radical is fully quenched in the presence of a physiological concentration of thiols. In the 2'-deoxyguanosine system, the C5' radical undergoes intramolecular attack onto the C8-N7 double bond of guanine leading ultimately to the 5',8-cyclo-2'-deoxyguanosine derivative. The cyclization of the 2'-deoxyguanosin-5'-yl radical occurs with a rate constant of ca. 1x10(6) s-1 and is highly stereoselective affording only the (5'S) diastereomer. PMID- 17425369 TI - Preparation and Diels-Alder chemistry of 4-vinylimidazoles. AB - Various 4-vinylimidazole derivatives have been prepared from the corresponding 4 iodoimidazoles or from urocanic acid. Several methods for the elaboration of these vinylimidazoles and their Diels-Alder reactions are reported. All of the vinylimidazoles prepared in the course of this study react with N-phenylmaleimide quite readily with mild thermal activation providing a single cycloadduct, in most cases the initial, nonaromatic adduct. With more electron rich substrates, there is a tendency for these initial cycloadducts to undergo aromatization, ene reaction, and oxidation although this can be circumvented to a large extent by the choice of reaction conditions. Limited reactions were observed with other dienophiles, providing the expected cycloadducts in most cases, although an abnormal adduct was obtained in one case with dimethyl acetylene dicarboxylate. These substrates also participate in regioselective Diels-Alder reactions with monoactivated dienophiles, but require fairly forcing conditions, thus only providing the aromatized cycloadducts in modest yields. An investigation of substituent effects at the 2-position of the imidazole moiety was undertaken, in which electron-donating and weakly electron-withdrawing substituents are tolerated. In addition, several substrates with terminally substituted vinyl moieties have been investigated. PMID- 17425370 TI - Efficient functionalizations of heteroatom-bridged calix[2]arene[2]triazines on the larger rim. AB - Heteroatom-bridged dichlorinated calix[2]arene[2]triazines, which were synthesized from the fragment coupling reactions of cyanuric chloride and various aromatic dinucleophiles, are a unique type of platform for the construction of functional macrocyclic host molecules. Utilizing a very convenient and straightforward nucleophilic displacement reaction of dichlorinated tetraoxacalix[2]arene[2]triazine by various chelating group-containing amines, a number of functionalized tetraoxacalix[2]arene[2]triazines on the larger rim were efficiently synthesized in good yields. The resulting tetraoxacalix[2]arene[2]triazines armed with two 2,2'-bi(pyridinyl)amino or two bis(2-pyridinemethyl)amino groups selectively formed 1:1 complexes with Cu2+ ion through most probably a chelating interaction effect. PMID- 17425371 TI - Aminolyses of aryl diphenylphosphinates and diphenylphosphinothioates: effect of modification of electrophilic center from P=O to P=S. AB - A kinetic study is reported for aminolysis of aryl diphenylphosphinothioates (2a i). The phosphinothioates 2a-i are less reactive than aryl diphenylphosphinates (1a-i), the oxygen analogues of 2a-i, regardless of the basicity of the leaving aryloxides or the attacking amines. The Yukawa-Tsuno plot for the reactions of 2b i with piperidine exhibits good linearity with a small r value (r=0.28), indicating that the leaving group departs at the rate-determining step with a small degree of bond fission. Reactions of 2,4-dinitrophenyl diphenylphosphinothioate (2a) with alicyclic secondary amines result in a good linear Bronsted-type plot with betanuc=0.52, implying that the reactions proceed through a concerted mechanism. The betanuc value determined for the reactions of 2a is slightly larger than that reported for the corresponding reactions of 2,4 dinitrophenyl diphenylphosphinate (1a, i.e., betanuc=0.38), suggesting that reactions of 2a proceed through a tighter transition state (TS) than that of 1a. The reaction of 2a with piperidine exhibits a ca. 0.4 kcal/mol more favorable enthalpy of activation (DeltaH) than that of 1a. On the contrary, the entropy of activation at 25.0 degrees C (TDeltaS) is ca. 1.5 kcal/mol more unfavorable for the reaction of 2a than for that of 1a. This result supports the proposal that the reaction of 2a proceeds through a tighter TS than that of 1a and explains why 2a-i are less reactive than 1a-i. PMID- 17425372 TI - Remote p-doping of InAs nanowires. AB - We report on remote p-type doping of InAs nanowires by a p-doped InP shell grown epitaxially on the core nanowire. This approach addresses the challenge of obtaining quantitative control of doping levels in nanowires grown by the vapor liquid-solid (VLS) mechanism. Remote doping of III-V nanowires is demonstrated here with the InAs/InP system. It is especially challenging to make p-type InAs wires because of Fermi level pinning around 0.1 eV above the conduction band. We demonstrate that shielding with a p-doped InP shell compensates for the built-in potential and donates free holes to the InAs core. Moreover, the off-current in field-effect devices can be reduced up to 6 orders of magnitude. The effect of shielding critically depends on the thickness of the InP capping layer and the dopant concentration in the shell. PMID- 17425373 TI - Photon storage with nanosecond switching in coupled quantum well nanostructures. AB - Photon storage with nanosecond switching was implemented with indirect excitons in coupled quantum well nanostructures. The storage and release of photons was controlled by the gate voltage pulses. The nanosecond write and readout times were much shorter than the storage time, which reached microseconds. Furthermore, the presented control of excitons on a time scale much shorter than the exciton lifetime demonstrates the feasibility of studying excitons in in situ controlled electrostatic traps. PMID- 17425376 TI - The Azzopardi phenomenon. PMID- 17425377 TI - Decidual vasculopathic-like lesion in endometrial ablation. PMID- 17425379 TI - Protocol for the examination of specimens from pediatric patients with hepatoblastoma. PMID- 17425380 TI - Basal/myoepithelial cells in chronic sinusitis, respiratory epithelial adenomatoid hamartoma, inverted papilloma, and intestinal-type and nonintestinal type sinonasal adenocarcinoma: an immunohistochemical study. AB - CONTEXT: The pathogenesis of respiratory epithelial adenomatoid hamartoma (REAH) and inverted papilloma (IP) is poorly understood, especially compared with sinonasal adenocarcinoma (SNAC). One feature of malignant glandular lesions is loss of the basal/myoepithelial layer. The immunophenotype of the basal/myoepithelial layer has not been fully examined in benign glandular lesions of the sinonasal tract. OBJECTIVE: To examine benign and malignant glandular lesions in the sinonasal tract for the immunophenotype of basal/myoepithelial cells, proliferation index, and cytokeratin and intestinal differentiation profiles. DESIGN: Sinonasal adenocarcinoma (intestinal-type adenocarcinoma [ITAC] and nonintestinal type adenocarcinoma [non-ITAC]), REAH, IP, and chronic sinusitis (CS) were stained for cytokeratin (CK) 7, CK20, 34betaE12, CDX-2, p63, Ki-67, smooth muscle actin (SMA), S100 protein, and calponin. RESULTS: Basal/myoepithelial cells in CS and REAH were positive for p63 and 34betaE12 but negative for SMA, S100 protein, and calponin. Proliferative activity was localized to the compartment containing p63-positive cells. Inverted papilloma demonstrated broad areas staining for p63 and 34betaE12, with intermediate proliferative activity in these areas. Sinonasal adenocarcinoma had the highest Ki-67 labeling index, and p63-positive SNACs had higher proliferation indices than p63-negative SNACs. REAH, IP, CS, and most SNACs expressed CK7. Only SNAC expressed CK20. Sixty percent of morphologic ITACs expressed CDX-2. CONCLUSIONS: Basal/myoepithelial cells in CS and REAH should be considered basal and not myoepithelial cells. In benign lesions, proliferative activity is limited to the compartments with p63 staining. In SNAC and IP, p63 expression correlates with proliferation index. REAH, IP, and CS share similar immunoprofiles (CK7+, CK20-, and CDX-2-), contrasting with SNAC (CK7+, CK20+/-, CDX-2-/+). PMID- 17425381 TI - Hemangiomas and angiosarcomas of the breast: diagnostic utility of cell cycle markers with emphasis on Ki-67. AB - CONTEXT: Vascular tumors comprise a minor subgroup of tumors arising in the breast and represent variants of hemangiomas and angiosarcomas. Diagnostic challenges may arise when differentiating hemangiomas from types I and II angiosarcomas. Ki-67 expression has been used as an adjunct to distinguish between benign and malignant lesions exhibiting histologic overlap at various anatomic sites. OBJECTIVE: To investigate the utility of Ki-67 and other cell cycle regulatory proteins (S-phase kinase-associated protein 2 [Skp2], p27, and cyclin D1) in the differential diagnosis of mammary vascular lesions. DESIGN: Thirty-four vascular tumors (21 hemangiomas and 13 angiosarcomas) of the breast were studied. The Ki-67 index and immunoreactivity for Skp2, p27, and cyclin D1 were determined in each case. Appropriate statistical methods were used. RESULTS: The mean value of Ki-67 index was statistically different when comparing hemangiomas and angiosarcomas (P < .001). Angiosarcomas were typically positive for Skp2, whereas hemangiomas were negative (P < .001). Sensitivity and specificity cutoffs for Ki-67 index to distinguish hemangiomas from angiosarcomas showed a candidate cutoff point of 175. The mean values of Ki-67 of low-grade angiosarcomas were significantly different from all hemangiomas (P < .001) and also different from the subset of atypical hemangiomas (P = .02). Sensitivity and specificity cutoffs for Ki-67 index to distinguish all hemangiomas from low-grade angiosarcomas showed a candidate cutoff point between 150 and 175. Among angiosarcomas, positivity for Ki-67 was inversely related to that of p27 but not to Skp2 or cyclin D1. This was also true among hemangiomas. CONCLUSIONS: Ki-67 index can be used as a diagnostic tool to distinguish between benign and malignant vascular lesions of the breast. This can be particularly helpful in cases of histologic overlap such as low-grade angiosarcoma and hemangioma. PMID- 17425382 TI - Adequacy of pathology resident training for employment: a survey report from the Future of Pathology Task Group. AB - CONTEXT: The recent change in accreditation requirements for anatomic pathology and clinical pathology residency training from 5 to 4 years and the rapid advances in technologies for pathology services have sparked a renewed debate over the adequacy of pathology residency training. In particular, perceived deficiencies in training have been declared from a variety of sources, both in the form of recent editorial opinions and from surveys of community hospital pathologist employers in 1998, 2003, and 2005 by Dr Richard Horowitz. OBJECTIVE: To obtain more comprehensive data on the perceptions of strengths and weaknesses in pathology residency training. DESIGN: The College of American Pathologists conducted a survey of potential pathology employers (senior College of American Pathologists members, members designated as head of group, and members of the Association of Directors of Anatomic and Surgical Pathology). Also surveyed were recent graduates of pathology residency programs, who were identified as being junior members of the College of American Pathologists, were recent recipients of certification from the American Board of Pathology, or were contacted through their directors of pathology residency programs. RESULTS: There were 559 employer respondents, of whom 384 were responsible for hiring and/or supervising new pathologists. There were 247 recent graduates of pathology residency training programs who responded. From the employers' standpoint, the majority expressed overall satisfaction with recent graduates, but almost one third of employers indicated that new hires had a major deficiency in a critical area. Specific areas of deficiency were clinical laboratory management and judgment in ordering special stains and studies. In addition, one half of employers agreed that more guidance and support for newly trained pathologists is needed now than was required 10 years ago. Academic employers generally were more satisfied than private sector employers. Newly trained pathologists did not appear to be inappropriately overconfident in their abilities. In addition, their perceptions of those specific areas in which they are most and least prepared are very similar to the ratings provided by employers. On average, newly trained pathologists' ratings of their own preparedness are highest for specific aspects of general pathology and anatomic pathology, and lowest for specific aspects of clinical pathology and administration. In selecting new pathologists, employers perceived medical knowledge and interpersonal skills as the most important discriminating applicant characteristics. When new employees were asked why they thought they were offered their position, the discriminating qualifications cited most often were academic background and training, as well as completion of a fellowship and subspecialty training. CONCLUSIONS: It is our hope that the results of this survey can be used as input for further discussions and recommendations for training of pathology residents so as to further advance the ability of pathologists to provide quality patient care upon their graduation from training. PMID- 17425383 TI - Kimura disease. AB - Kimura disease is a benign rare chronic inflammatory disorder of unknown etiology that involves the lymph nodes and subcutaneous tissue of the head and neck regions. Elevated serum immunoglobulin E levels and peripheral blood eosinophilia are also common. This disease is most common in middle-aged Asian men. Although the etiology is unknown, it most probably represents an aberrant chronic immune response. Treatment for Kimura disease includes surgical resection and regional or systemic steroid therapy. Cytotoxic therapy and radiation have also been utilized. The disease has an excellent prognosis, although it may recur locally. PMID- 17425384 TI - Double immunohistochemical staining with MUC4/p53 is useful in the distinction of pancreatic adenocarcinoma from chronic pancreatitis: a tissue microarray-based study. AB - CONTEXT: Immunohistochemical stains have been used for the distinction of pancreatic adenocarcinoma from chronic pancreatitis. OBJECTIVE: To determine if a double stain for MUC/p53 improved specificity and sensitivity for distinction of pancreatic adenocarcinoma from chronic pancreatitis by comparing maspin, mucin 4 (MUC4), p53, Smad4, and the double stain MUC4/p53. DESIGN: Seventy-four pancreatic adenocarcinomas and 19 chronic pancreatitis cases were retrieved from archival files. Tissue cores were arrayed to create a tissue microarray of 2-mm cores. Sections were stained with antibodies against maspin, MUC4, p53, and Smad4. Additionally, a 2-color, double stain for MUC4 and p53 was developed and evaluated. Five percent or greater staining in either of the cores was considered positive. Intensity (0, 1, 2) and extent (%) of tumor cells staining was also determined. RESULTS: The sensitivity for distinction of pancreatic adenocarcinoma from chronic pancreatitis with maspin, MUC4, p53, and Smad4 was 90%, 77%, 60%, and 63%, respectively; the specificity was 67%, 78%, 88%, and 88%, respectively. When MUC4 and p53 were combined in a double stain, and positive staining for either considered a positive result, the sensitivity increased to 96% but specificity was 73%. When immunoreactivity for both antibodies was necessary for a positive result, sensitivity fell to 39% but specificity was 100%. No correlation was found between intensity or extent of staining with any of the individual stains and tumor differentiation. CONCLUSION: The double immunohistochemical stain for MUC4/p53 can be a useful diagnostic tool in conjunction with the hematoxylin-eosin-stained section for pancreatic adenocarcinoma, particularly when limited tumor is available for multiple stains. PMID- 17425385 TI - Serum levels of matrix metalloproteinase 3 (stromelysin 1) for monitoring synovitis in rheumatoid arthritis. AB - CONTEXT: Matrix metalloproteinase 3 (MMP-3) is expressed in synovial tissues and involved in cartilage destruction in rheumatoid arthritis and osteoarthritis. OBJECTIVE: To study whether measurement of MMP-3 serum concentrations is useful to monitor the activity of rheumatoid synovitis. DESIGN: Levels of MMP-3 in serum and synovial tissue samples obtained from 29 rheumatoid arthritis patients and 20 osteoarthritis patients were measured by the 1-step sandwich enzyme immunoassay system. RESULTS: Levels of MMP-3 in the serum and synovial samples were significantly higher in rheumatoid arthritis than in osteoarthritis (P < .001), and the levels correlated directly with each other (r = 0.712, P < .001; N = 49). Immunohistochemistry demonstrated almost exclusive localization of MMP-3 to the lining cells in rheumatoid synovium. The immunoreactivity correlated directly with the scores of synovial inflammatory cell infiltration (r = 0.606, P < .001; n = 29) and the MMP-3 levels in the synovial tissues (r = 0.564, P = .001; n = 29) and those in the serum samples (r = 0.529, P = .003; n = 29) in rheumatoid arthritis. Levels of MMP-3 in rheumatoid serum samples dropped to low values at 1 and 2 weeks after total knee arthroplasty, while the levels of C-reactive protein increased at 1 week and the erythrocyte sedimentation rate and counts of white blood cells and platelets were unchanged at 1 and 2 weeks postoperative. CONCLUSIONS: Our results demonstrate that MMP-3 levels in the serum of rheumatoid arthritis patients correlate with the levels produced by the synovial lining cells and suggest that the activity of rheumatoid synovitis can be monitored by measuring serum levels of MMP-3. PMID- 17425386 TI - Goblet cell mimickers in esophageal biopsies are not associated with an increased risk for dysplasia. AB - CONTEXT: Identification of intestinal-type goblet cells (ITGCs) in hematoxylin eosin-stained sections of esophageal biopsies is essential for the diagnosis of Barrett metaplasia. However, we have seen cases diagnosed as Barrett metaplasia based solely on cells that pose morphologic similarity to ITGCs on hematoxylin eosin staining or stain positive with Alcian blue. OBJECTIVE: To determine the clinical significance of goblet cell mimickers. DESIGN: Initial biopsies from 78 patients with original diagnosis of Barrett metaplasia negative for dysplasia and a mean follow-up of 72 months were reviewed and reclassified into 3 categories: (1) ITGCs, (2) goblet cell mimickers, or (3) neither. Sections from available paraffin blocks were stained with Alcian blue at pH 2.5. The presence of the different types of cells and positive Alcian blue staining were correlated with each other and evaluated for their significance as predictors of progression to dysplasia. RESULTS: Goblet cell mimickers were present in 35 cases and were associated with ITGCs in the same biopsy in 23 (66%) of these cases. Intestinal type goblet cells were present in 56 cases, and the remaining 10 cases, although called Barrett on the original report, did not show either ITGCs or goblet cell mimickers. Only the presence of ITGCs was associated with significant risk for dysplasia (P = .008). Positive Alcian blue staining was not associated with a significant risk for dysplasia. CONCLUSIONS: Our results indicate that the diagnosis of Barrett metaplasia should be rendered with confidence only when ITGCs are identified on routine hematoxylin-eosin-stained sections. PMID- 17425387 TI - Ensuring timely completion of type and screen testing and the verification of ABO/Rh status for elective surgical patients. AB - CONTEXT: A blood bank can provide compatible blood for an elective surgical procedure, provided a blood sample is received by the laboratory with sufficient time to allow pretransfusion testing and acquire enough compatible red blood cell units. With the push for same-day admission surgical procedures, a patient's pretransfusion blood sample is often collected the morning of surgery. However, if blood is needed, compatible units might not be immediately available. OBJECTIVE: To define and improve the process of completing presurgical/preadmission type and screen testing and verifying the ABO/Rh status of scheduled surgical patients before they receive a transfusion. STUDY DESIGN: A list of surgical procedures that might necessitate blood transfusion was created. A checklist was used to ensure that the preoperative clinic nurse collects a baseline pretransfusion blood sample for type and screen testing from patients scheduled for a listed procedure. A new pretransfusion specimen was received on the day of surgery, if needed, so that a current specimen would be available for compatibility testing and to verify the accuracy of the patient's ABO/Rh status in case blood was requested. RESULTS: During the 1-year study period, 666 patients qualified for baseline type and screen testing. Cholecystectomy was the most commonly scheduled surgery. In 99% of cases, a baseline type and screen specimen was received in the laboratory at least 1 day before surgery. The interval between the preoperative clinic visit and date of surgery varied from same day (6 patients) to 3 months. CONCLUSION: Timely receipt of a presurgical specimen for type and screen testing and verification of a patient's ABO/Rh status can be ensured when clinical services collaborate and when the hospital blood utilization committee provides oversight to improve compliance. PMID- 17425388 TI - Thyroid transcription factor 1 immunohistochemistry as an intraoperative diagnostic tool at frozen section for distinction between primary and secondary lung tumors. AB - CONTEXT: Intraoperative distinction between primary and metastatic carcinomas in the lung at frozen section remains problematic. OBJECTIVE: To assess the value and practicality of immunohistochemistry for thyroid transcription factor 1 at the time of intraoperative frozen section. DESIGN: Thirty-three patients presented with either a solitary pulmonary mass or 2 pulmonary masses and a history of carcinoma in a different organ. In addition to routine frozen section for assessment of tumor type, we looked for expression of thyroid transcription factor 1, using the EnVision system with abridged methodology. RESULTS: Ten cases were positive for thyroid transcription factor 1, which was confirmed on subsequent paraffin sections. Nine of these were confirmed as primary pulmonary adenocarcinomas, but 1 case proved to be a rare false-positive metastatic colonic carcinoma. Twenty-three cases were negative on frozen section and reported as favoring metastatic disease. In all cases, additional immunohistochemical data increased diagnostic confidence, but particularly in cases of positive primary pulmonary tumors and in cases with disease metastatic from sites other than the large bowel. The average time in addition to that of the basic frozen section was 24 minutes per test with a cost of 32 pounds sterling (US$57). CONCLUSIONS: Frozen section immunohistochemistry for thyroid transcription factor 1 shows specificity and sensitivity similar to those seen for formalin-fixed tissues and is feasible within the time frame of a thoracotomy. Diagnostic confidence is increased, especially with positive primary pulmonary tumors. However, its practice should be properly planned within an operative procedure as liberal usage will likely have significant staff and cost implications. PMID- 17425389 TI - Clinical laboratory specimen rejection--association with the site of patient care and patients' characteristics: findings from a single health care organization. AB - CONTEXT: Programs to track laboratory quality have reported aggregated specimen rejection rates ranging from 0.30% to 0.83%. Because the performance of the laboratory, rather than errors, has been the focus, reasons for specimen rejection or demographic characteristics of individuals at risk for specimens of poor quality may not be fully understood. OBJECTIVE: To calculate the proportions of rejected specimens stratified by point of collections and demographic information of patients. DESIGN: Retrospective cross-sectional study. Data were retrieved from the intrainstitutional electronic databases. RESULTS: The proportions of specimens that were rejected in the emergency department and inpatient services were 2-fold and more than 5-fold higher, respectively, than for the outpatient services. Assessment of data by patients' ethnic heritages yielded no significant differences among African Americans (0.38%), Caucasians (0.38%), or "Others" (0.35%) in the outpatient services (P = .07). In the emergency department, the proportions of rejected specimens for African Americans (2.24%) were almost twice that of Caucasians (1.39%) and 30% higher than for Others (1.70%). A similar finding was observed for the inpatient services. CONCLUSIONS: The effect of ethnicity on the proportions of rejected specimens was significant for samples that were collected in the emergency department and inpatient services, even after adjusting for the total number of specimens. A constellation of factors, that is, disease severity and seriousness, practice of blood sample collection, and lesser proficiency of the nursing staff in phlebotomy may be reasons for this observation. However, the likelihood of differential care, although unlikely, cannot be refuted by the present data. PMID- 17425390 TI - MUC4 expression in non-small cell lung carcinomas: relationship to tumor histology and patient survival. AB - CONTEXT: Mucin 4 (MUC4) is a high-molecular-weight membrane-bound glycoprotein that is expressed in the foregut before epithelial differentiation. It is found in normal adult airway epithelium, non-small cell lung carcinoma (NSCLC) and in other human malignancies independent of mucus secretion. Although its tissue distribution has been studied, its utility in predicting prognosis in NSCLC is unknown. OBJECTIVE: To evaluate the relationship between MUC4 overexpression and long-term survival in patients with NSCLC. DESIGN: Immunohistochemical staining for MUC4 was performed on formalin-fixed, paraffin-embedded tissue samples from 343 cases of NSCLC arranged in a high-density tissue microarray. Information about long-term survival and tumor stage was collected for all patients. Semiquantitative assessment of MUC4 staining was correlated with survival (Kaplan Meier analysis). RESULTS: MUC4 was frequently expressed in adenocarcinomas (151/187 [81%]), squamous cell carcinomas (69/ 88 [78%]), adenosquamous carcinomas (6/8 [75%]), and large cell carcinomas (33/60 [55%]). High levels of expression (combined score, 2+/3+) for MUC4 were more characteristic of adenocarcinomas (126/187 [68%]) and adenosquamous carcinomas (6/8 [75%]) than of squamous cell carcinomas (46/88 [52%]) and large cell carcinomas (17/60 [28%]) (P < .001). In patients with stage I and II adenocarcinoma, there was a trend toward longer patient survival with higher levels of MUC4 immunoreactivity compared with lower levels (P = .11). CONCLUSION: MUC4 expression is common in pulmonary adenocarcinomas and may indicate a more favorable prognosis in early-stage adenocarcinomas. PMID- 17425391 TI - Distinguishing prostatic from colorectal adenocarcinoma on biopsy samples: the role of morphology and immunohistochemistry. AB - CONTEXT: Poorly differentiated carcinoma on prostate or colorectal biopsy can occasionally present a diagnostic challenge in determining tumor source especially in locally advanced colorectal carcinoma (CRCa) or prostate carcinoma (PCa). Such determination can affect prognosis and therapy. OBJECTIVE: To evaluate the role of morphology and immunohistochemistry in the previously mentioned setting. DESIGN: Surgical pathology and consultation records. Hematoxylin-eosin sections were reviewed in 16 cases (11 PCa, 5 CRCa). Immunohistochemistry for 9 markers was performed in 15 cases. RESULTS: Dirty necrosis, seen in 5 (100%) of 5 CRCa and 2 (18%) of 11 PCa cases, and the presence of columnar cells with basal nuclei, seen in 5 (100%) of 5 CRCa and 1 (9%) of 11 PCa cases, appear to be the most useful morphologic parameters. Immunohistochemistry confirmed the value of prostate-specific antigen (PSA), CDX2, cytokeratin (CK) 20, and beta-catenin in the differential of CRCa (0% PSA+, 60% CDX2+, 80% CK20+, and 100% beta-catenin+) versus PCa (80% PSA+, 0% CDX2+, 10% CK20+, and 0% beta-catenin+). P501S had a similar sensitivity as PSA in detecting PCa (80%). Two (20%) of 10 PCa cases were positive for 1 of the 2 markers but not the other. P501S was negative in all 5 cases of CRCa. CONCLUSIONS: P501S is a useful marker in this setting when included together with PSA, CDX2, CK20, and beta-catenin. P501S labels a subset of PCa cases that are negative for PSA. Dirty necrosis and/or columnar cells with basal nuclei could also be of help. PMID- 17425392 TI - Questionable role of human herpesviruses in the pathogenesis of Kikuchi disease. AB - CONTEXT: Kikuchi disease is a self-limiting febrile lymphadenopathy characterized by a patchy area of apoptosis. Kikuchi disease is thought to be caused by a virus, but this has not been clearly demonstrated. Human herpesviruses 6 and 7 (HHV-6 and HHV-7) are lymphotropic viruses that can induce apoptosis in infected lymphocytes. Recently, HHV-8 was reported to be a possible etiologic agent of Kikuchi disease. OBJECTIVE: To investigate the incidence of HHV-6, HHV-7, and HHV 8 infection in patients with Kikuchi disease. DESIGN: Seventy archival tissue specimens (from 50 Kikuchi disease cases and 20 control cases) were tested for the presence of HHV-6 and HHV-7 using a nested polymerase chain reaction, and for the presence of HHV-8 using single-step polymerase chain reaction. Immunohistochemistry for HHV-8 expression was carried out in those cases in which HHV-8 was detected using polymerase chain reaction. RESULTS: Of the 50 cases with Kikuchi disease, 21 (42%) were HHV-6 positive and 32 (64%) were HHV-7 positive. Eight (40%) of the 20 control cases were HHV-6 positive and 9 (45%) were HHV-7 positive. Both HHV-6 and HHV-7 were detected in 15 (30%) of the cases with Kikuchi disease and in 3 (15%) of the control cases. Three (6%) of the 50 cases of Kikuchi disease were HHV-8 positive but revealed no positive cells on immunohistochemical analysis for HHV-8. Human herpesvirus 8 was not expressed in any of the control cases. CONCLUSIONS: There was no association between the presence of HHV-6 or HHV-7 and Kikuchi disease. Because the HHV-8 genome but not protein was detected in a small proportion of the cases of Kikuchi disease, its potential causative role in this disease should be determined by further studies. PMID- 17425393 TI - Leadership and management training for residents and fellows: a curriculum for future medical directors. AB - CONTEXT: Management of laboratories and pathology practices is increasingly complex. Residents and fellows in laboratory medicine and pathology need more structured curricula in leadership and management (L&M) training to function as medical and laboratory directors. OBJECTIVE: To define a curriculum that provides basic competency in L&M for residents and fellows in pathology. DESIGN: A year long formal L&M course included didactic lectures, interactive sessions, case scenarios, team-building exercises, formal team presentations (capstone project), and precourse and postcourse assessment of L&M knowledge. The curriculum meets requirements of American College of Graduate Medical Education and supports goals for leadership training of the College of American Pathologists. Participants evaluated (5-point scale) the content and speakers of all sessions. Trainees were evaluated after considering postcourse examination results, quality of the capstone presentation, and a global assessment. RESULTS: The 5 non-capstone sessions received evaluation scores ranging from 4.4 (informatics) to 5 (L&M basics). Postcourse test scores showed significant improvement when compared with the pretest scores for the 2003-2004 and 2004-2005 trainee cohorts. CONCLUSIONS: Short-term results indicate that the course described improves trainee knowledge of L&M issues. PMID- 17425394 TI - The medicolegal aspect of error in pathology: a search of jury verdicts and settlements. AB - CONTEXT: Identifying medical errors is a topic of current attention. Among the various approaches is the study of medical malpractice cases. OBJECTIVE: To identify the most common medical errors involving the practice of pathology from a medicolegal perspective by analysis of published jury verdict and settlement reports. DESIGN: Search approximately 50 publications that gather jury verdict and settlement information using LexisNexis, an on-line searchable archive, for pathology-related cases. RESULTS: One hundred seventy-one legal cases were identified from 1988 through 2005. Nearly one-half involved surgical pathology; among the remainder, cytology cases slightly outnumbered those pertaining to clinical pathology. Among the surgical pathology cases and overall, based on this database, the most common reason for a medical malpractice lawsuit related to pathology was the alleged missed diagnosis of melanoma on a skin biopsy specimen. Less commonly, the surgical pathology cases involved breast biopsy specimens, gynecological specimens, lung, genitourinary system, technical or preanalytic errors (eg, mixed-up specimens), soft tissue, hematopathology, head and neck, gastrointestinal/hepatobiliary system, or thyroid. Among the 48 cases related to cytology, 37 involved false-negative Papanicolaou smears. Less common were cases related to fine-needle aspirates of the breast or thyroid or cytology specimens of the lung. Among the 36 cases involving clinical pathology, 32 related to the blood bank--usually transfusion-acquired human immunodeficiency virus infection. CONCLUSIONS: These data are in agreement with other publications as to the most frequent causes of medical malpractice allegations related to pathology. As these issues are addressed, the number of errors should decrease. Studying the jury verdict and settlements data may provide additional insight into medical errors and patient safety. PMID- 17425395 TI - Interference of Asian, American, and Indian (Ashwagandha) ginsengs in serum digoxin measurements by a fluorescence polarization immunoassay can be minimized by using a new enzyme-linked chemiluminescent immunosorbent or turbidimetric assay. AB - CONTEXT: Ginsengs are widely used by the general population. These herbs interfere with serum digoxin measurement using the fluorescence polarization immunoassay. OBJECTIVE: To assess potential interference of different ginsengs (Asian, American, and Indian, also known as Ashwagandha) in vitro and in vivo in a mouse model by using a new enzyme-linked chemiluminescent immunosorbent digoxin assay and an existing turbidimetric assay. Comparisons were made with the fluorescence polarization immunoassay. DESIGN: Aliquots of drug-free serum pools were supplemented with ginseng and apparent digoxin concentrations were measured using enzyme-linked chemiluminescent immunosorbent digoxin assay, turbidimetric assay, and fluorescence polarization immunoassay digoxin assays. Mice were fed with different ginseng preparations and apparent digoxin concentrations were measured 1 and 3 hours later. In a separate experiment, aliquots of serum digoxin pools were further supplemented with ginsengs and the serum digoxin concentrations were measured again. RESULTS: A significant apparent digoxin concentration was observed both in vitro and in vivo using the fluorescence polarization immunoassay, but no apparent digoxin concentration was observed using enzyme-linked chemiluminescent immunosorbent digoxin assay and turbidimetric assay. No interference was observed with enzyme-linked chemiluminescent immunosorbent digoxin assay and turbidimetric assay when digoxin serum pools were further supplemented with various ginsengs. CONCLUSIONS: It was concluded that both enzyme-linked chemiluminescent immunosorbent and turbidimetric digoxin assays are free from ginseng interferences. PMID- 17425396 TI - Significance of margin and extent of dysplasia in loop electrosurgery excision procedure biopsies performed for high-grade squamous intraepithelial lesion in predicting persistent disease. AB - CONTEXT: High-grade squamous intraepithelial lesions (cervical intraepithelial neoplasia 2 and 3) are commonly treated with loop electrosurgery excision procedure (LEEP) biopsies. OBJECTIVE: To highlight the significance of positive margins and extent of positive margins of the cervical LEEP biopsies in predicting the persistence of high-grade squamous intraepithelial lesion and to provide suggestions for reporting margins in cervical LEEP biopsies. DESIGN: The pathology files at the University of Arkansas for Medical Sciences were searched for cervical intraepithelial neoplasia 2 and 3 treated by LEEP biopsy from 1990 to 2001. RESULTS: A total of 489 LEEP biopsy specimens were retrieved and reviewed; 270 patients had follow-up within 1 year. The biopsy specimens of 110 patients showed positive endocervical margins. One hundred sixty specimens had negative ectocervical-endocervical margins. Follow-up of 54% of the cases with initial positive margins showed residual high-grade squamous intraepithelial lesions. This association was even greater when multiple blocks showed positive endocervical margins and in cases with positive deep margins. On the other hand, a negative margin predicted ability to completely remove the lesion in 95% of patients. CONCLUSION: This study reiterates the significance of the evaluation of the margin, even in samples that were received as multiple fragments. Reporting of LEEP biopsy findings should include the extent of the dysplasia, the status of the ectocervical-endocervical margin, and the status of the deep margin. PMID- 17425397 TI - Perineurioma: a distinctive and underrecognized peripheral nerve sheath neoplasm. AB - CONTEXT: Perineuriomas are benign peripheral nerve sheath neoplasms composed of perineurial cells with characteristic immunohistochemical and ultrastructural features. They have been traditionally classified into two main types according to their location--intraneural and extraneural--and overlap histologically with many other tumors, which may be diagnostically challenging to general surgical pathologists. OBJECTIVE: To review the clinical, morphologic, immunohistochemical, ultrastructural, cytogenetic, and molecular genetic aspects of perineurioma, as well as to discuss its clinicopathologic variants and differential diagnosis. DATA SOURCES: English-language literature published between 1966 and 2005 was reviewed. CONCLUSIONS: The correct identification of perineuriomas is important to avoid unnecessary overtreatment. The histologic diagnosis should be confirmed through immunohistochemical studies (including epithelial membrane antigen, S100 protein, and more recently described antibodies such as claudin-1 and GLUT1) or electron microscopy. Cytogenetic and molecular genetic studies are still of limited value for the diagnosis of perineuriomas but may play a fundamental role in excluding important differential diagnoses and also in helping elucidate the biology of these poorly known neoplasms. PMID- 17425398 TI - Adenoid cystic/basal cell carcinoma of the prostate: review and update. AB - CONTEXT: Although most prostate carcinomas are of the conventional acinar type, unusual variants have been reported. Adenoid cystic/basal cell carcinoma of the prostate is a rare tumor with distinctive histopathologic features. There are only a few publications in the literature concerning the diagnosis, treatment, and prognosis of this neoplasm. OBJECTIVE: To review current literature together with the clinical, pathologic, and immunohistochemical features of adenoid cystic/basal cell carcinoma of the prostate and offer a practical approach to the diagnosis--including the differential diagnosis--of this neoplasm in surgical pathologic specimens. DATA SOURCES: Adenoid cystic/basal cell carcinoma of the prostate is composed of infiltrating basaloid cells forming dilated acinar and cribriform spaces with luminal basementlike material. Differentiation of adenoid cystic/basal cell carcinoma from basal cell hyperplasia and cribriform pattern of acinar adenocarcinoma may be difficult. The use of cytokeratin 34betaE12 and prostate-specific antigen can help in difficult cases. Most cases are indolent, but metastasis has been documented in a few cases. CONCLUSIONS: Various histologic and immunohistochemical features are helpful in recognizing adenoid cystic/basal cell carcinoma of the prostate. This is a rare subtype of prostate cancer and correct diagnosis is important because of the unique clinical and biological features and the implications for treatment and prognosis. PMID- 17425399 TI - Metastatic renal cell carcinoma to hemangioblastoma in von Hippel-Lindau disease. AB - A case of metastatic renal cell carcinoma (RCC) to a capillary hemangioblastoma (HAB) of the central nervous system in a 52-year-old woman with von Hippel-Lindau (vHL) syndrome is described. We review the literature on metastatic RCC to HAB, summarize the histologic and immunohistochemical features that can distinguish between the 2 tumors, and comment on the significance of such a finding in terms of the clinical diagnosis of vHL. We found the expression of CAM 5.2, RCC antigen, and CD10 to be strong in RCC and absent in HAB and, conversely, staining with Leu-7, neural cell adhesion molecule, and inhibin-alpha was present in HAB but weak or absent in RCC. These antibodies can be used to differentiate these entities, provided one is astute in recognizing the possibility of their coexistence. PMID- 17425400 TI - Epithelial marker-negative desmoplastic small round cell tumor with atypical morphology: definitive classification by fluorescence in situ hybridization. AB - Desmoplastic small round cell tumor typically presents with abundant desmoplastic stroma containing nested primitive round cells bearing a polyphenotypic immunohistochemical profile. Lesions with minimal classic morphology pose a formidable diagnostic challenge. The current case represents one such example, arising as a large abdominal-pelvic mass in a 17-year-old female patient. The tumor was composed of a monomorphous population of small round cells lining microcystic structures and forming pseudoacini and fine anastomosing trabeculae and cords. The stroma was abundantly myxoid with only occasional thick desmoplastic septa. The tumor cells were variably immunopositive for vimentin, desmin, smooth muscle actin, synaptophysin, neuron-specific enolase, Bcl-2 and WT1 (nuclear); epithelial markers were negative. The definitive diagnosis of desmoplastic small round cell tumor was rendered with the demonstration of the characteristic EWS-WT1 gene fusion by fluorescence in situ hybridization. The current case emphasizes the utility of fluorescence in situ hybridization to demonstrate EWS-WT1 gene fusion in desmoplastic small round cell tumor with nonclassic morphologic and immunohistochemical features to avoid potential misdiagnosis. PMID- 17425401 TI - Disseminated coccidioidomycosis in pregnancy. AB - Coccidioidomycosis is a fungal infection contracted through the inhalation of airborne spores, which are most frequently present in desert areas of the southwestern United States and Mexico. Primary immune response to infection is by T(H)1, a subset of helper T cells. Although pulmonary symptoms are most common, hematogenous systemic spread can also occur. Pregnancy is a well-noted risk factor for disseminated Coccidioides infection. The objective of this review is to provide an overview of coccidioidomycosis and to review immunologic and hormonal factors that increase risk of dissemination in pregnancy. Dissemination may occur more frequently in pregnant patients than in nonpregnant women because of shifts in T-cell immunity, changes in cytokine production, and increased hormone levels. There is disagreement regarding the precise incidence of systemic spread in pregnancy, but most sources agree that risk is substantially increased and vigilance must be high in patients with exposures in endemic areas. PMID- 17425403 TI - Signature-based small molecule screening identifies cytosine arabinoside as an EWS/FLI modulator in Ewing sarcoma. AB - BACKGROUND: The presence of tumor-specific mutations in the cancer genome represents a potential opportunity for pharmacologic intervention to therapeutic benefit. Unfortunately, many classes of oncoproteins (e.g., transcription factors) are not amenable to conventional small-molecule screening. Despite the identification of tumor-specific somatic mutations, most cancer therapy still utilizes nonspecific, cytotoxic drugs. One illustrative example is the treatment of Ewing sarcoma. Although the EWS/FLI oncoprotein, present in the vast majority of Ewing tumors, was characterized over ten years ago, it has never been exploited as a target of therapy. Previously, this target has been intractable to modulation with traditional small-molecule library screening approaches. Here we describe a gene expression-based approach to identify compounds that induce a signature of EWS/FLI attenuation. We hypothesize that screening small-molecule libraries highly enriched for FDA-approved drugs will provide a more rapid path to clinical application. METHODS AND FINDINGS: A gene expression signature for the EWS/FLI off state was determined with microarray expression profiling of Ewing sarcoma cell lines with EWS/FLI-directed RNA interference. A small-molecule library enriched for FDA-approved drugs was screened with a high-throughput, ligation-mediated amplification assay with a fluorescent, bead-based detection. Screening identified cytosine arabinoside (ARA-C) as a modulator of EWS/FLI. ARA C reduced EWS/FLI protein abundance and accordingly diminished cell viability and transformation and abrogated tumor growth in a xenograft model. Given the poor outcomes of many patients with Ewing sarcoma and the well-established ARA-C safety profile, clinical trials testing ARA-C are warranted. CONCLUSIONS: We demonstrate that a gene expression-based approach to small-molecule library screening can identify, for rapid clinical testing, candidate drugs that modulate previously intractable targets. Furthermore, this is a generic approach that can, in principle, be applied to the identification of modulators of any tumor associated oncoprotein in the rare pediatric malignancies, but also in the more common adult cancers. PMID- 17425404 TI - TREM2-transduced myeloid precursors mediate nervous tissue debris clearance and facilitate recovery in an animal model of multiple sclerosis. AB - BACKGROUND: In multiple sclerosis, inflammation can successfully be prevented, while promoting repair is still a major challenge. Microglial cells, the resident phagocytes of the central nervous system (CNS), are hematopoietic-derived myeloid cells and express the triggering receptor expressed on myeloid cells 2 (TREM2), an innate immune receptor. Myeloid cells are an accessible source for ex vivo gene therapy. We investigated whether myeloid precursor cells genetically modified to express TREM2 affect the disease course of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. METHODS AND FINDINGS: EAE was induced in mice by immunization with a myelin autoantigen. Intravenous application of TREM2-transduced bone marrow-derived myeloid precursor cells at the EAE peak led to an amelioration of clinical symptoms, reduction in axonal damage, and prevention of further demyelination. TREM2-transduced myeloid cells applied intravenously migrated into the inflammatory spinal cord lesions of EAE-diseased mice, showed increased lysosomal and phagocytic activity, cleared degenerated myelin, and created an anti-inflammatory cytokine milieu within the CNS. CONCLUSIONS: Intravenously applied bone marrow-derived and TREM2-tranduced myeloid precursor cells limit tissue destruction and facilitate repair within the murine CNS by clearance of cellular debris during EAE. TREM2 is a new attractive target for promotion of repair and resolution of inflammation in multiple sclerosis and other neuroinflammatory diseases. PMID- 17425405 TI - Aphid thermal tolerance is governed by a point mutation in bacterial symbionts. AB - Symbiosis is a ubiquitous phenomenon generating biological complexity, affecting adaptation, and expanding ecological capabilities. However, symbionts, which can be subject to genetic limitations such as clonality and genomic degradation, also impose constraints on hosts. A model of obligate symbiosis is that between aphids and the bacterium Buchnera aphidicola, which supplies essential nutrients. We report a mutation in Buchnera of the aphid Acyrthosiphon pisum that recurs in laboratory lines and occurs in field populations. This single nucleotide deletion affects a homopolymeric run within the heat-shock transcriptional promoter for ibpA, encoding a small heat-shock protein. This Buchnera mutation virtually eliminates the transcriptional response of ibpA to heat stress and lowers its expression even at cool or moderate temperatures. Furthermore, this symbiont mutation dramatically affects host fitness in a manner dependent on thermal environment. Following a short heat exposure as juveniles, aphids bearing short allele symbionts produced few or no progeny and contained almost no Buchnera, in contrast to aphids bearing symbionts without the deletion. Conversely, under constant cool conditions, aphids containing symbionts with the short allele reproduced earlier and maintained higher reproductive rates. The short allele has appreciable frequencies in field populations (up to 20%), further supporting the view that lowering of ibpA expression improves host fitness under some conditions. This recurring Buchnera mutation governs thermal tolerance of aphid hosts. Other cases in which symbiont microevolution has a major effect on host ecological tolerance are likely to be widespread because of the high mutation rates of symbiotic bacteria and their crucial roles in host metabolism and development. PMID- 17425406 TI - Structural and chemical profiling of the human cytosolic sulfotransferases. AB - The human cytosolic sulfotransfases (hSULTs) comprise a family of 12 phase II enzymes involved in the metabolism of drugs and hormones, the bioactivation of carcinogens, and the detoxification of xenobiotics. Knowledge of the structural and mechanistic basis of substrate specificity and activity is crucial for understanding steroid and hormone metabolism, drug sensitivity, pharmacogenomics, and response to environmental toxins. We have determined the crystal structures of five hSULTs for which structural information was lacking, and screened nine of the 12 hSULTs for binding and activity toward a panel of potential substrates and inhibitors, revealing unique "chemical fingerprints" for each protein. The family wide analysis of the screening and structural data provides a comprehensive, high level view of the determinants of substrate binding, the mechanisms of inhibition by substrates and environmental toxins, and the functions of the orphan family members SULT1C3 and SULT4A1. Evidence is provided for structural "priming" of the enzyme active site by cofactor binding, which influences the spectrum of small molecules that can bind to each enzyme. The data help explain substrate promiscuity in this family and, at the same time, reveal new similarities between hSULT family members that were previously unrecognized by sequence or structure comparison alone. PMID- 17425407 TI - Object-oriented echo perception and cortical representation in echolocating bats. AB - Echolocating bats can identify three-dimensional objects exclusively through the analysis of acoustic echoes of their ultrasonic emissions. However, objects of the same structure can differ in size, and the auditory system must achieve a size-invariant, normalized object representation for reliable object recognition. This study describes both the behavioral classification and the cortical neural representation of echoes of complex virtual objects that vary in object size. In a phantom-target playback experiment, it is shown that the bat Phyllostomus discolor spontaneously classifies most scaled versions of objects according to trained standards. This psychophysical performance is reflected in the electrophysiological responses of a population of cortical units that showed an object-size invariant response (14/109 units, 13%). These units respond preferentially to echoes from objects in which echo duration (encoding object depth) and echo amplitude (encoding object surface area) co-varies in a meaningful manner. These results indicate that at the level of the bat's auditory cortex, an object-oriented rather than a stimulus-parameter-oriented representation of echoes is achieved. PMID- 17425408 TI - A dominant, recombination-defective allele of Dmc1 causing male-specific sterility. AB - DMC1 is a meiosis-specific homolog of bacterial RecA and eukaryotic RAD51 that can catalyze homologous DNA strand invasion and D-loop formation in vitro. DMC1 deficient mice and yeast are sterile due to defective meiotic recombination and chromosome synapsis. The authors identified a male dominant sterile allele of Dmc1, Dmc1(Mei11), encoding a missense mutation in the L2 DNA binding domain that abolishes strand invasion activity. Meiosis in male heterozygotes arrests in pachynema, characterized by incomplete chromosome synapsis and no crossing-over. Young heterozygous females have normal litter sizes despite having a decreased oocyte pool, a high incidence of meiosis I abnormalities, and susceptibility to premature ovarian failure. Dmc1(Mei11) exposes a sex difference in recombination in that a significant portion of female oocytes can compensate for DMC1 deficiency to undergo crossing-over and complete gametogenesis. Importantly, these data demonstrate that dominant alleles of meiosis genes can arise and propagate in populations, causing infertility and other reproductive consequences due to meiotic prophase I defects. PMID- 17425409 TI - HPA-1a/b(PlA1/A2,Zwa/b): the odyssey of an alloantigen system. PMID- 17425410 TI - Neonatal alloimmune thrombocytopenia: a 50-year story. PMID- 17425411 TI - Platelet additive solutions: current status. PMID- 17425412 TI - ABO and platelet transfusion therapy. PMID- 17425413 TI - Scott Murphy's contribution in the early years of posttransfusion purpura: a remembrance. PMID- 17425414 TI - Differences in ABO antibody levels among blood donors: a comparison between past and present Japanese, Laotian, and Thai populations. AB - Passively transfused blood group antibodies cause clinical problems. High titers of anti-A and anti-B seem to be one reason for hemolytic transfusion reactions and for ABO HDN. In Japan, anti-A and anti-B titers notably decreased in the 15 years between 1986 and 2001. At present, titers of more than 100,as measured using the saline method, are rare. Differences in the level of anti-A and anti-B among ethnic populations have been reported; these differences were found to be the result of environmental factors rather than hereditary factors. In the present study, the anti-A and anti-B titers of random donors in three Asian populations are compared. In Thailand, the IgM anti-A and anti-B titers are low and are similar to the Japanese titers reported in 2001, but the IgG anti-A and anti-B titers are high and are similar to the Japanese titers reported in 1986. In the Lao People's Democratic Republic, both the IgM and the IgG anti-A and anti B titers are high and are similar to those reported in Japan in 1986. In addition, anti-A and anti-B titers of different sex donors and of various age groups were also compared. High titers were found in 8.8 percent of the female donors in the younger than 30 age group and in 36.7 percent of the female donors in the older than 50 age group. PMID- 17425416 TI - Long-lasting poliovirus-neutralizing antibodies among Argentinean population immunized with four or five oral polio vaccine doses 1 month to 19 years previously. AB - The persistence of poliovirus-neutralizing antibodies was investigated in 297 individuals residing in Argentina who had completed the vaccination cycle with four or five oral polio vaccine (OPV) doses 1 mo to 19 yr before this study. Seropositivity for the three polio types in individuals who had received four OPV doses remained high and stable, showing rates not less than 94.6, 98.2, and 91.1% for types 1, 2, and 3, respectively, for a period of at least 6 yr. Almost identical rates were found in children who completed a vaccination schedule of five OPV doses 1 to 2 yr earlier. However, humoral immunity to poliovirus types 3 and 1 declined significantly 9 and 17 yr, respectively, after the booster dose had been administered; in contrast, type 2 immunity remained fairly stable during the 19-yr study period. Overall, geometric mean titer values for poliovirus types 1 and 2 were higher than those for poliovirus type 3. This is likely a result of low initial poliovirus type 3 antibody titers that eventually fell below the limits of detection at later time points. The results indicate that although antibody titers primed by OPV decline over time, they are remarkably long-lived, immunity to poliovirus types 1 and 2 being more prevalent than that against type 3 at late intervals postvaccination. PMID- 17425417 TI - Serologic prevalence of coxsackievirus group B in Greece. AB - Coxsackieviruses are human enteroviruses, which have been associated with myocarditis/pericarditis and sudden death. In one investigation (Spanakis N, Manolis EN, Tsakris A, Tsiodras S, Panagiotopoulos T, Saroglou G, and Legakis NJ: J Clin Pathol 2005;58:357-360), a cluster of cases of fatal myocarditis in Greece was linked to coxsackievirus B3. The information from this investigation prompted us to study serologically the prevalence of coxsackieviruses B throughout Greece. Sera were obtained from 506 healthy blood donors from various transfusion centers, covering the entire country. All sera were tested for the presence of IgG and IgM antibodies, using ELISAs with various antigenic specificities: (1) heat-denatured coxsackievirus type B1 and B5 virions, (2) a synthetic peptide from the N terminus of the VP1 protein of coxsackievirus B3, and (3) a synthetic peptide from the N terminus of the VP1 protein of coxsackievirus B4. Sera positive for IgG antibodies against coxsackieviruses B1/B5, B3, and B4 were detected in 6.7 to 21.6% of the individuals tested in the various regions of Greece. Statistical analysis revealed that the highest prevalence of IgG antibodies against coxsackieviruses B1/B5 was found in blood donors from Crete (p = 0.025), whereas the highest prevalence against coxsackievirus B4 was detected in blood donors from Athens (p = 0.01). IgM antibodies against coxsackievirus B were detected at low percentage, less than 5%, with no significant viral preference for particular geographic regions. The preference of anti coxsackievirus IgG antibodies for particular geographic regions could be potentially related to the previously reported clustering of cases of insulin dependent diabetes mellitus and myocarditis in Athens and Crete, respectively. PMID- 17425418 TI - CCR2 regulates development of Theiler's murine encephalomyelitis virus-induced demyelinating disease. AB - Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease, a murine model for multiple sclerosis, involves recruitment of T cells and macrophages to the CNS after infection. We hypothesized that CCR2, the only known receptor for CCL2, would be required for TMEV-induced demyelinating disease development because of its role in macrophage recruitment. TMEV-infected SJL CCR2 knockout (KO) mice showed decreased long-term clinical disease severity and less demyelination compared with controls. Flow cytometric data indicated that macrophages (CD45(high) CD11b(+) ) in the CNS of TMEV-infected CCR2 KO mice were decreased compared with control mice throughout disease. CD4(+) and CD8(+) T cell percentages in the CNS of TMEV-infected control and CCR2 KO mice were similar over the course of disease. There were no apparent differences between CCR2 KO and control peripheral immune responses. The frequency of interferon-gamma producing T cells in response to proteolipid protein 139-151 in the CNS was also similar during the autoimmunity stage of TMEV-induced demyelinating disease. These data suggest that CCR2 is important for development of clinical disease by regulating macrophage accumulation after TMEV infection. PMID- 17425419 TI - Progressive biliary destruction is independent of a functional tumor necrosis factor-alpha pathway in a rhesus rotavirus-induced murine model of biliary atresia. AB - Rhesus rotavirus (RRV)-inoculated neonatal BALB/c mice develop an immune-mediated inflammation of extra- and intrahepatic bile ducts that progresses to biliary obstruction and death by 3 wk of age. Livers of diseased animals demonstrate increased numbers of T lymphocytes with elevated expression of helper T cell type 1 (Th1) cytokines at 1 wk, which transitions to increased numbers of macrophages and high expression of the proinflammatory cytokine tumor necrosis factor (TNF) alpha by 2 wk. We employed both pharmacologic and genetic approaches for attenuation of TNF-alpha to determine whether it plays a causal role in injury. First, RRV-inoculated BALB/c mice were subjected to multiple treatments with either the TNF receptor I (TNF-RI)-Fc fusion protein etanercept or neutralizing antibodies to mouse TNF-alpha. Also, TNF-RI(-/-) mice were injected with RRV in the same manner as wild-type mice. In all cases, TNF inhibition did not reduce the severity or incidence of disease. Survival curves of mice given blocking agents were similar to those of control RRV-inoculated mice, and survival of challenged TNF-RI(-/-) mice was worse than that of wild-type mice, likely because of the prolonged presence of infectious RRV. In all experimental groups, markers of disease were unchanged from those of control mice. In summary, although RRV inoculated BALB/c mice have highly elevated expression of TNF-alpha, this cytokine does not play an obligate role in disease progression. PMID- 17425420 TI - Local and systemic human papillomavirus type 6b-specific cellular immune responses in patients with recurrent genital warts. AB - Anogenital warts are a common clinical manifestation of genital infection with human papillomavirus type 6b (HPV-6b). Accumulating data indicate that an effective cellular immune response is required for the control of HPV infections. However, in a minority of patients there is a high rate of recurrence of wart lesions. We report the characterization of both local and systemic HPV-specific cellular immune responses in three patients with a history of recurrent genital warts. Although the patients had chronic recurrent wart lesions, we have shown that each had both detectable intralesional and peripheral HPV-specific T lymphocytes. Interestingly, the lesion-infiltrating T cells were specific for only one HPV antigen, focusing on only a few epitopes. Conversely, the T cells derived from peripheral blood recognized a broader range of HPV antigens. The characteristics of the HPV-specific cellular immunity that we have shown in these patients may be indicative of a failure to mount an effective response against the virus. This would be consistent with the chronic nature of the disease in these specific individuals. These observations could be relevant to the design and immunomonitoring of immunotherapeutic vaccines for persistent HPV infections. PMID- 17425421 TI - Cellular immune responses in acute hepatitis E virus infection to the viral open reading frame 2 protein. AB - Hepatitis E virus (HEV) causes acute viral hepatitis and is endemic in the developing world. Few data are available on cellular immune responses in HEV infection. Using flow cytometry, we studied the frequencies of peripheral blood CD4(+) /CD8(+) T cells secreting interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-4 in 21 patients with acute hepatitis E and 18 healthy controls, after stimulation with the HEV capsid (ORF2) protein. Cytokine levels in serum specimens and culture supernatants of ORF2-stimulated peripheral blood mononuclear cells (PBMCs) were estimated in enzyme-linked immunosorbent assays. In addition, cytokine mRNA transcripts were measured in PBMCs by reverse transcription-polymerase chain reaction. In patients with acute hepatitis E, although the total CD4(+) population was expanded, the proportions of CD4(+)/CD69(+) and CD8(+) /CD69(+) cells producing IFN-gamma, TNF-alpha, and IL-4 in response to HEV ORF2 stimulation were unchanged. However, IFN-gamma levels in the supernatants and IFN-gamma mRNA transcripts in cells were elevated in ORF2 stimulated PBMCs in acute hepatitis E; levels of IL-2 or TNF-alpha were unchanged. Our findings suggest that CD4(+) IFN-gamma-secreting cells, which do not belong either to the helper T cell type 1 or type 2 phenotype, as is the case with natural killer T cells, may be involved in the pathogenesis of hepatitis E. Further, the limited immune reactivity we detected in peripheral blood cells may be related to the sequestration of immune events to the intrahepatic compartment, which is the major disease site. PMID- 17425422 TI - Increased interleukin-17 production both in helper T cell subset Th17 and CD4 negative T cells in human immunodeficiency virus infection. AB - Interleukin (IL)-17 is produced mainly by activated CD4(+) T cells, currently known as Th17. Human immunodeficiency virus (HIV) pathogenesis leads to CD4(+) T cell depletion. This is the first report of IL-17 in HIV infection. We assessed IL-17 expression in the CD4(+) T cells (Th17) of 40 asymptomatic HIV-infected treatment-naive patients compared with 40 HIV-seronegative volunteers. Peripheral blood mononuclear cells (PBMCs), with/without phorbol myristate acetate (PMA)/ionomycin stimulation, were stained with CD3, CD4, IL-17, and interferon (IFN)-gamma antibodies and analyzed by four-color flow cytometry. Both groups had comparable baseline data, except for age (mean+/-SD): 36 +/- 9 versus 30 +/- 9 yr (p= 0.001), CD4(+) T cell counts (median): 218 versus 623 cells/microL (p < 0.0001), CD8(+) T cell counts (median): 875.5 versus 382.5 cells/microL ((p) < 0.0001), and CD4(+)/CD8(+) cell ratios (median): 0.225 versus 1.45 (p< 0.0001). Without stimulation, the percentages of IL-17(+) CD3(+) CD4() and IL-17(+) CD3(+) CD4() cells among HIV-seropositive and -seronegative volunteers (median) were as follows: 0.68 versus 0.12% (p< 0.0001) and 0.92 versus 0.09% (p< 0.0001), respectively. With PMA/ionomycin stimulation, the percent IL-17 expression in CD4(+) cells (median) was 1.45 versus 0.65 (p< 0.0001) and in CD4() T cells it was 1.0 versus 0.12 (p< 0.0001). In conclusion, HIV infection is associated with a significant increase in IL-17 production in both CD4(+) and CD4() T cells in peripheral blood. IL-17 expression was further inducible by PMA/ionomycin stimulation in vitro only in CD4(+) T cells. The roles of IL-17 and Th17 in HIV viral replication and immunopathogenesis are under further investigation. PMID- 17425423 TI - Distinct pathways for signaling maturation in macrophages and dendritic cells after infection with paramyxovirus simian virus 5. AB - Professional antigen-presenting cells are critical components of both the innate and adaptive immune responses. Although dendritic cells (DCs) are generally thought to be the primary activators of naive T cells, macrophages have also been shown to fulfill this role. As with DCs, the capacity to induce optimal activation of T cells requires that macrophages undergo a process that results in the increased expression of costimulatory molecules, such as CD40, CD80, and CD86, and the production of cytokines. In this study we analyzed the effect of infection of macrophages generated from BALB/c mice with the paramyxovirus simian virus 5 (SV5). Here we have shown that bone marrow-derived macrophages (BMMs) are not productively infected at any multiplicity of infection tested. Analysis of activation markers revealed that SV5-infected BMMs robustly upregulated CD40 and modestly upregulated CD86, but did not upregulate the expression of CD80. Further, SV5-infected BMMs secreted low levels of interferon-beta and interleukin (IL)-12p40, but high levels of tumor necrosis factor-alpha and IL-6. Intriguingly, upregulation of these molecules on BMMs, unlike our previous results using bone marrow-derived dendritic cells, was not dependent on live virus. These findings provide evidence that different professional antigen presenting cells can detect and respond to virus via distinct mechanisms. PMID- 17425424 TI - Restricted and selective tropism of a Venezuelan equine encephalitis virus derived replicon vector for human dendritic cells. AB - Dendritic cells (DCs) consist of heterogeneous phenotypic populations and have diverse immunostimulatory functions dependent on both lineage and functional phenotype, but as exceptionally potent antigen-presenting cells, they are targets for generating effective antigen-specific immune responses. A promising replicon particle vector derived from Venezuelan equine encephalitis virus (VEE) has been reported to transduce murine footpad DCs. However, the receptive DC subset, the degree of restriction for this tropism, and the extent of conservation between rodents and humans have not been well characterized. Using fresh peripheral blood DCs, mononuclear cells, monocyte-derived macrophages, and monocyte-derived DCs, our results demonstrate conservation of VEE replicon particle (VRP) tropism for DCs between humans and rodents. We observed that a subset of immature myeloid DCs is the target population, and that VRP-transduced immature DCs retain intact functional capacity, for example, the ability to resist the cytopathic effects of VRP transduction and the capacity to acquire the mature phenotype. These studies support the demonstration of selective VRP tropism for human DCs and provide further insight into the biology of the VRP vector, its parent virus, and human DCs. PMID- 17425425 TI - Innate immune responses to replication of porcine reproductive and respiratory syndrome virus in isolated Swine alveolar macrophages. AB - Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease caused by a positive RNA strand arterivirus. PRRS virus (PRRSV) interacts primarily with lung macrophages. Identifying the genetic components involved in host resistance/susceptibility would represent an important step forward in the design of disease control programs. In this study, alveolar macrophages derived from five commercial pig lines were used to study the innate immune response to PRRSV infection in vitro. Analysis by flow cytometry has demonstrated that bronchial alveolar lavage fluid (BALF) preparations were almost exclusively composed of alveolar macrophages and that the pigs tested were free from infection. Macrophages from the Landrace line showed significantly reduced virus replication and poor growth of PRRSV during 30 h of infection. By 72 h, PRRSV viral load was down to 2.5 log(10) TCID(50) compared with an average of 5 log(10) TCID(50) for the other breeds tested. These observations suggest that factors intrinsic to the Landrace breed may be responsible for this reduced or delayed response to PRRSV. Preliminary investigation suggests that the PRRSV coreceptor, sialoadhesin, may not be responsible for the Landrace macrophage phenotype as its abundance and localisation were comparable in all the breeds. Strikingly, we found that the reduced or delayed growth of PRRSV was temporally associated with high levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-8 mRNA accumulation and substantial reduction of secretion of IL-8, suggesting a key contributory role for cytokine synthesis and secretion during the innate immune response to PRRSV infection. PMID- 17425426 TI - Neonatal ovine pulmonary dendritic cells support bovine respiratory syncytial virus replication with enhanced interleukin (IL)-4 And IL-10 gene transcripts. AB - The lung microenvironment is constantly exposed to microorganisms and particulate matter. Lung dendritic cells (DCs) play a crucial role in the uptake and processing of antigens found within the respiratory tract. Respiratory syncytial virus (RSV) is a common respiratory tract pathogen in children that induces an influx of DCs to the mucosal surfaces of the lung. Using a neonatal lamb model, we examined the in vivo permissiveness of DCs to RSV infection, as well as overall cell surface changes and cytokine responses of isolated lung DCs after bovine RSV (BRSV) infection. We report that isolated lung DCs and alveolar macrophages support BRSV replication. Isolated lung DCs were determined to be susceptible to BRSV infection as demonstrated by quantification of BRSV non structural protein 2 mRNA. BRSV infection induced an initial upregulation of CD14 expression on lung DCs, but by 5 d postinfection expression was similar to that on control cells. No significant changes in CD80/86 or MHC class I expression were seen on lung DCs after BRSV infection. Low to moderate expression of MHC class II and DEC-205 was detected by day 5 postinfection. Initially, on day 3 postinfection, lung DCs from BRSV-infected lambs had decreased endocytosis of fluorescein isothiocyanate (FITC)-ovalbumin (OVA). The amount of FITC-OVA endocytosed by lung DCs isolated on day 5 postinfection was similar to that of controls. The most interesting observation was the induction of immunomodulatory interleukin (IL)-4 and IL-10 cytokine gene transcription in lung DCs and alveolar macrophages after in vivo infection with BRSV. Overall, these findings are the first to demonstrate that neonatal lung DCs support in vivo BRSV replication and produce type II cytokines after viral infection. PMID- 17425427 TI - Immunologic, virologic, and neuropsychologic responses in human immunodeficiency virus-infected children receiving their first highly active antiretroviral therapy regimen. AB - Our objective was to measure the early dynamics, evolution, and durability over 96 wk of immunologic responses in children receiving their first highly active antiretroviral therapy (HAART) regimen. The study was designed as a prospective, single-arm study. Twelve human immunodeficiency virus (HIV)-infected children (median age, 11.8 yr) were enrolled. All subjects received stavudine, nevirapine, and ritonavir. Serial measurements included HIV viral load, lymphocyte subsets, thymic volume by computed tomography (CT), neurocognitive testing, and brain CT. Baseline median CD4(+) T cell count was 589 cells/mm(3) , viral load was 3.9 log(10) HIV RNA copies/mL, and thymic volume was 16.3 cm(3) . Ten children had an undetectable viral load at week 48. Eight maintained an undetectable viral load at 96 wk. The median increase in absolute CD4(+) T cell count was 225 cells/mm(3) by week 48, and 307 cells/mm(3) by week 96. The median increase in naive (CD45RA(+) CD62L(+) ) CD4(+) T cells was 133 cells/mm(3) by week 48, and 147 cells/mm(3) by week 96. The median number of naive CD8(+) T cells increased from 205 to 284 cells/mm(3) by week 24; this increase was sustained to week 96. The number of B cells increased and was associated with a decrease in immunoglobulin levels. The number of natural killer cells was stable. There were no significant changes in thymic volume. Most children exhibited stable cognitive function over the course of the study. We conclude that, in this cohort of relatively immunocompetent HIV-infected children, an initial HAART regimen was associated with rapid and sustained increases in total CD4(+) T cells, in naive CD4(+) and CD8(+) T cells, and in B cells through 96 wk. PMID- 17425428 TI - Using interferon-alpha to block expression of cellular ribosome subunit S24 variant 2 in human fibroblasts inhibits translation of the poliovirus genome. AB - Studies about the proteins induced by interferon (IFN-)-alpha stimulation have provided some data on their mechanism of antiviral effect. These proteins were confirmed to contribute to antiviral functions. In this study, IFN-alpha stimulation of human fibroblasts was shown to induce the inhibition of S24 variant 2 (a structural component of the ribosomal small subunit) at the mRNA and protein levels, implying a possible antiviral mechanism for IFN-alpha in human fibroblasts. The delay of poliovirus replication by IFN-alpha was partially compensated for by S24 variant 2 expressed in pcDNA vector-transfected cells, and the interference RNA of S24 variant 2 was able to induce mimetically, to some extent, this poliovirus replication delay. These observations revealed that S24 variant 2 could be involved in the antiviral effects of IFN-alpha in human fibroblasts. PMID- 17425429 TI - Subcellular localization and characterization of G protein-coupled receptor homolog from lymphocystis disease virus isolated in China. AB - G protein-coupled receptors (GPCRs) constitute a large superfamily involved in various types of signal transduction pathways, and play an important role in coordinating the activation and migration of leukocytes to sites of infection and inflammation. Viral GPCRs, on the other hand, can help the virus to escape from host immune surveillance and contribute to viral pathogenesis. Lymphocystis disease virus isolated in China (LCDV-C) contains a putative homolog of cellular GPCRs, LCDV-C GPCR. In this paper, LCDV-C GPCR was cloned, and the subcellular localization and characterization of GPCR protein were investigated in fish cells. LCDV-C GPCR encoded a 325 amino acid peptide, containing a typical seven transmembrane domain characteristic of the chemokine receptors and a conserved DRY motif that is usually essential for receptor activation. Transient transfection of GPCR-EGFP in fathead minnow (FHM) cells and epithelioma papulosum cyprini (EPC) cells indicated that LCDV-C GPCR was expressed abundantly in both the cytoplasm and nucleoplasm. Transient overexpression of GPCR in these two cells cannot induce obvious apoptosis. FHM cells stably expressing GPCR showed enhanced cell proliferation and significant anchorage-independent growth. The effects of GPCR protein on external apoptotic stimuli were examined. Few apoptotic bodies were observed in cells expressing GPCR treated with actinomycin D (ActD). Quantitative analysis of apoptotic cells indicated that a considerable decrease in the apoptotic fraction of cells expressing GPCR, compared with the control cells, was detected after exposure to ActD and cycloheximide. These data suggest that LCDV-C GPCR may inhibit apoptosis as part of its potential mechanism in mediating cellular transformation. PMID- 17425430 TI - Dendritic cell-based vaccine strategy against human immunodeficiency virus clade C: skewing the immune response toward a helper T cell type 2 profile. AB - Given the continued spread of human immunodeficiency virus (HIV)-1 worldwide, developing efficient vaccine strategies against HIV-1 is a key task. We tested the safety and immunogenicity of a multicomponent, cell-based vaccine that consisted of antigen-expressing apoptotic bodies with or without autologous dendritic cells (DCs). The vaccine strategy involved transfection of human 293T cells with codon-optimized DNA vectors expressing env of HIV1084i, a newly transmitted pediatric HIV-1 clade C strain; SHIV89.6P tat; and SIVmac239 gag protease. Apoptotic bodies were generated by heat shock and ultraviolet irradiation and mixed either with mouse DCs (DC-cell vaccine) or given directly (cell-only vaccine) to BALB/c mice for initial priming; boosts consisted of apoptotic bodies only. The immunogens were well tolerated with or without DCs. Compared with the cell-only vaccine, the DC-cell vaccine induced higher antibody titers against all three antigens, whereas virus-specific cytotoxic T lymphocyte responses were equally strong in both groups. Iso-type analysis of viral antigen specific antibodies revealed a skewing toward helper T type 2 responses induced by the DC-cell vaccine but not by the cell-only vaccine. In summary, both vaccine strategies were safe and induced cellular as well as humoral antiviral immunity; the DC-based approach had the advantage of significantly stronger antibody responses. PMID- 17425431 TI - Immunization with a synthetic multiepitope antigen induces humoral and cellular immune responses to hepatitis C virus in mice. AB - The immunogenicity of a synthetic multiepitope PCX3 antigen, which contains triple tandem repeats of five conserved epitopes from hepatitis C virus (HCV) polyprotein, was studied in BALB/c mice given three intraperitoneal injections of antigen with Freund's adjuvant. Both a strong antibody response and specific cytotoxic T lymphocytes were induced. The specific anti-PCX3 IgG was able to bind HCV particles from hepatitis C patient sera by incubation overnight. In particular, in transgenic mice with chimeric human livers, anti-PCX3 antibody was able to lower the viral load in two of five mice and to eliminate HCV infection in three of five mice by 2 wk after inoculation with HCV-positive serum from patients. These results indicated that the synthetic multiepitope PCX3 antigen elicits a potent humoral and cellular immune response against HCV. PMID- 17425432 TI - Evaluation of human immunodeficiency virus type 1-infected Chinese patients treated with highly active antiretroviral therapy for two years. AB - The purpose of this study was to evaluate the long-term efficacy and safety of nevirapine in combination with didanosine and stavudine in the treatment of human immunodeficiency virus (HIV)1-infected Chinese patients in routine clinical practice. The study, from April 2003 to May 2005, with follow-up through 24 mo, was conducted at the Department of Infectious Diseases, Second Xiangya Hospital, Central-South University in Changsha, Hunan Province, China. Twenty-seven HIV1 infected patients received didanosine, stavudine, and nevirapine. Information from case notes regarding age, sex, side effects, viral load, naive and memory T cells, and CD4(+) and CD8(+) T cell count at baseline, 3, 6, 12, 18, and 24 mo was collected and analyzed. Virologic suppression, defined as an HIV RNA concentration of less than 50 copies/mL at months 3, 6, 12, 18, and 24, was considered the main outcome measure. Of 27 patients, 17 were men with a mean age 33.5 yr. The mean baseline viral load was 5.15 log copies/mL and the mean CD4(+) cell count was 185 cells/dL. Of 27 patients, 3 patients discontinued study medication; treatment was changed, because of side effects, from didanosine (ddI), stavudine (d4T), and nevirapine (NVP) to zidovudine, lamivudine, and NVP for 24 patients who had completed 24 mo of treatment with ddI, d4T, and NVP; and viral load suppression was attained in 17 patients (70.8%) at 12 mo, in 14 patients (58.3%) at 18 mo, and in 13 patients (56.6%) at 24 mo. The CD4 T cell count increased by 114 cells/microL (mean, 299 cells/microL) after 12 mo of treatment and by 132 cells/microL (mean, 317 cells/microL) after 24 mo of treatment. Naive T cells and memory cells also increased in number, but at a slower rate. Activated (CD38(+)) CD8(+) T cells were elevated at baseline (67.7%) and declined by month 24 (49.7%), but did not reach normal levels. We conclude that a regimen of NVP with ddI and d4T provided durable suppression of plasma viral load in HIV-infected patients, with significant improvement in the CD4 cell count, and can be well tolerated by patients with HIV-1 infection. PMID- 17425433 TI - Pretreatment with recombinant human vascular endothelial growth factor reduces virus replication and inflammation in a perinatal lamb model of respiratory syncytial virus infection. AB - Vascular endothelial growth factor (VEGF) is increasingly recognized as a perinatal regulator of lung maturation and surfactant protein expression. Preterm and young infants are at increased risk for pulmonary immaturity characterized by insufficient surfactant production as well as increased risk for severe manifestations of respiratory syncytial virus (RSV) infection. Innate immune components including surfactant proteins A and D, and beta-defensins have putative antimicrobial activity against pulmonary pathogens including RSV. Our hypothesis was that recombinant human VEGF (rhVEGF) pretreatment therapy would decrease RSV disease in the perinatal lamb RSV model. Newborn lambs were pretreated with rhVEGF, betamethasone, or saline and then inoculated with bovine RSV or sterile medium. Tissues were collected 5 d postinoculation, corresponding to the initiation of severe lesions and peak viral replication. In RSV-infected lambs, rhVEGF therapy increased the mean daily body temperature, decreased airway neutrophil exudate, and reduced RSV replication compared with betamethasone or saline pretreatment. Furthermore, rhVEGF therapy significantly mitigated the RSV induced increase in surfactant protein A mRNA expression and decrease in surfactant protein D mRNA expression. In control (non-RSV-infected) lambs, pretreatment with rhVEGF increased sheep beta-defensin-1 (SBD1) mRNA expression, but no alteration in surfactant proteins A and D was detected. This novel study demonstrates that rhVEGF pretreatment mitigates RSV disease and, in addition, rhVEGF regulation of innate immune genes is dependent on RSV infection status. PMID- 17425434 TI - Inhibition of severe acute respiratory syndrome-associated coronavirus infection by equine neutralizing antibody in golden Syrian hamsters. AB - Equine anti-severe acute respiratory syndrome-associated coronavirus F(ab')(2) has been verified to protect mice from infection with severe acute respiratory syndrome-associated coronavirus (SARS-CoV). However, before potential clinical application, the antibody needs to be tested in as many animal models as possible to ensure its safety and efficiency. In this study, after verification by various methods that the golden Syrian hamster constitutes a model susceptible to SARS CoV infection, we confirmed that the antibody could protect animals completely from SARS-CoV infection in the preventive setting. More importantly, the antibody could reduce viral titers or copies by approximately 10(3)- to 10(4)-fold in animal lung after virus exposure, compared with negative control. These data provide further evidence to warrant clinical studies of this antibody in the treatment and prevention of SARS. PMID- 17425435 TI - Chemokine production predominates in human monocytes infected with Tula virus. AB - Many reports suggest the hypothesis of a complex immune response accompanying hantaviral infections. However, little is known about the immunopathogenesis of nonpathogenic hantaviruses, especially Tula virus (TULV). The aim of our study was to determine the cytokine/chemokine profile induced after the infection of human macrophages with TULV and the role of viral replication in this process. Also, we wanted to establish how the study of TULV is relevant to our previous study of pathogenic hantaviruses. We showed that TULV-infected macrophages produced chemokines (interleukin-8, macrophage chemoattractant protein-1, and macrophage inflammatory protein-1beta) important for recruiting inflammatory cells, whereas no significant changes were recorded in the tested cytokine levels. This property was not influenced by ultraviolet inactivation. There were some differences in chemokine production compared with our previous study with pathogenic hantaviruses. A possible explanation could be a different way of entering host cells found in the pathogenic and nonpathogenic hantaviruses and activation of different intracellular signaling pathways. PMID- 17425436 TI - CD4+ T cells play a pivotal role in the control of cowpox virus infection in mice. PMID- 17425438 TI - Pharmacodynamics and stability of subcutaneously infused glucagon in a type 1 diabetic Swine model in vivo. AB - BACKGROUND: The objective of this study was to determine the in vivo pharmacodynamics of glucagon and to test its glycemic effect over days by assessing its time course of activity and potency in a type 1 diabetic swine model. METHODS: Individual experiments were conducted in different pigs using glucagon preparations that were reconstituted on different days and stored at room temperature or near body temperature before usage. All experiments involved a subcutaneous bolus of glucagon to counter impending hypoglycemia induced by an earlier bolus of fast-acting insulin. Frequent blood-glucose measurements, using a standard in vitro hand-held meter, were taken during each experiment to track variations in blood-glucose concentration. RESULTS: Significant glucagon action was observed as early as 5 min after administration, as evidenced by an effective halt to declining blood glucose and a subsequent twofold rise in blood glucose after approximately 20 min. Results also indicate that the consumption of glucagon from the subcutaneous depot is substantially faster than that of fast acting insulin. Furthermore, no significant depreciation was observed in glucagon efficacy across aging preparations as old as 7 days. CONCLUSIONS: These results suggest profound utility of subcutaneous glucagon in a closed-loop glucose control system, especially since glucagon would provide an effective safeguarding measure to stave off impending hypoglycemia, an application where the rapid effect of subcutaneous glucagon is both serendipitous and essential. Despite the notion that the stability of glucagon in solution at room temperature is inferior to that of fast-acting insulin, its subcutaneous administration has promising prospects for long-term closed-loop ambulatory care. PMID- 17425439 TI - Type I diabetes mellitus in human and chimpanzee: a comparison of kyoto encyclopedia of genes and genomes pathway. AB - BACKGROUND: Diabetes is a worldwide medical problem and is a significant cause of morbidity and mortality. Type 1 diabetes results from the autoimmune destruction of insulin-producing beta cells in the pancreas. The identification of causative genes for the autoimmune disease type 1 diabetes in humans has made significant progress in recent years. Studies of pathways for type 1 diabetes in other living things can give useful information on the nature of type 1 diabetes. METHODS: Here, the author used a new pathway technology to compare type 1 diabetes mellitus in the human and the chimpanzee. RESULTS: According to the comparison, the mainframes of pathways are similar for both the human and the chimpanzee. CONCLUSIONS: These results can imply a close relation between the human and the chimpanzee. They also confirm usage of the chimpanzee model for studies of type 1 diabetes pathophysiology. PMID- 17425440 TI - Waist circumference identifies primary school children with metabolic syndrome abnormalities. AB - OBJECTIVE: This study was designed to assess whether waist circumference can predict metabolic syndrome abnormalities in primary schoolchildren. RESEARCH DESIGN AND METHODS: Of 5,103 children (2,526 males) 4-13 years old who underwent anthropometric measurements, 530 had more extensive testing. Body mass index (BMI), waist circumference, and blood pressure were determined in all subjects. The subgroup had Tanner stage, glucose, lipid profile, and insulin assays. The BMI of the 5,103 children was used to calculate our z scores. To determine which marker was a better predictor for metabolic syndrome, a receiver operating characteristic (ROC) curve was generated for BMI and waist circumference, with metabolic syndrome as the dichotomous variable. RESULTS: Over 530 children (8.7 +/- 2.4 years) 6% (n = 32) were obese (BMI >95(th) percentile; z BMI = 2.55), 13.6% (n = 72) were overweight (OW) (85(th) < BMI < 95(th) percentile; z BMI = 1.45), and 80.4% (n = 426) were non-OW (BMI <85(th) percentile; z BMI = - 0.14). Fifty-eight percent [95% confidence interval (CI) 53, 6], 22.8% (95% CI 19, 27), 15.5% (95% CI 12, 19), and 4.1% (95% CI 2, 6) were Tanner stage I, II, III, and IV, respectively. Metabolic syndrome was present in 9.4% overall, 6% of the non OW, 22.2% of the OW, and 31% in the obese group (P < 0.01). The differences between ROC areas were not significant (0.009) (95% CI -0.035 to 0.053; P = 0.679) for BMI and waist circumference. The optimal threshold for waist circumference percentile was 71.3 with a sensitivity and specificity of 58.9 (95% CI 48.4, 68.9) and 63.1 (95% CI 58.4, 67.7), respectively. CONCLUSIONS: Waist circumference and BMI predict metabolic syndrome abnormalities in children. Waist circumference > or =75(th) percentile could be the optimal threshold to predict metabolic syndrome in children. PMID- 17425441 TI - The HealthPia GlucoPack Diabetes phone: a usability study. AB - BACKGROUND: Type I diabetes is a common chronic disease of childhood. Both the growing influence of peers and the shifting away from parental influence have been implicated as prime elements contributing to poor glycemic outcomes in adolescents. Mobile technology that can be directed towards providing self management support and modifying potentially negative child parent interaction holds promise to improve control in adolescents with diabetes. METHODS: HealthPia, Inc. (Palisades Park, NJ) has developed a prototype system, the HealthPia GlucoPack Diabetes Monitoring System, which integrates a small blood glucose monitoring device into the battery pack of a cell phone. A pilot study used mixed quantitative and qualitative methods to evaluate user satisfaction with the integrated system, including the potential of the device to transmit self-monitoring data to a website for review and analysis by clinicians, parents, and patients. RESULTS: Adolescents in our study liked the integration of the two technologies and agreed that the glucometer was easy to use and that the tool was useful in the management of their diabetes. CONCLUSIONS: Future work will focus on the utilization of the diabetes phone as a component of a care delivery system for adolescents with diabetes, including involvement of the health care team and enhancement of the web services that support the use of the phone. PMID- 17425442 TI - Relative accuracy of the BD Logic and FreeStyle blood glucose meters. AB - BACKGROUND: The BD Logic((R)) (Becton, Dickinson and Co., Franklin Lakes, NJ) and FreeStyle((R)) (Abbott Diabetes Care, Alameda, CA) meters are used to transmit data directly to insulin pumps for calculation of insulin doses and to calibrate continuous glucose sensors as well as to monitor blood glucose levels. METHODS: The accuracy of the two meters was evaluated in two inpatient studies conducted by the Diabetes Research in Children Network (DirecNet). In both studies, meter glucose measurements made with either venous or capillary blood were compared with reference glucose measurements made by the DirecNet Central Laboratory at the University of Minnesota using a hexokinase enzymatic method. RESULTS: The BD Logic tended to read lower than the laboratory reference regardless of whether venous (median difference = -9 mg/dL) or capillary blood (median difference = -7 mg/dL) was used. This resulted in lower accuracy of the BD Logic compared with the FreeStyle meter based on the median relative absolute difference (RAD) for both venous blood (median RAD, 9% vs. 5%, P < 0.001) and capillary blood (median RAD, 11% vs. 6%, P = 0.008). The greatest discrepancy in the performance of the two meters was at higher reference glucose values. Accuracy was not significantly different when the reference was < or = 70 mg/dL. CONCLUSIONS: The BD Logic meter is less accurate than the FreeStyle meter. PMID- 17425443 TI - Relationship between interstitial and blood glucose in type 1 diabetes patients: delay and the push-pull phenomenon revisited. AB - BACKGROUND: Interpretation of glucose sensor results requires clarification of the relationship between interstitial (IG) and blood (BG) glucose. We examined the delay of IG upon BG change and reinvestigated the push-pull phenomenon in type 1 diabetes patients. The push-pull phenomenon postulates that IG shows a delayed increase but earlier decrease compared to BG. If so, postprandial sensor curves should have narrower peak widths than BG curves. METHODS: For both sensors a two-point calibration procedure was used. Delay was assessed by shifting combined fitted postprandial glucose sensor curves horizontally. The sensor and BG peak widths of the separately fitted curves were assessed and compared. Peak width was re-assessed for the microdialysis sensor using raw current values to rule out any calibration effect on the shape of the curve. The contribution of instrumental delay to the earlier reported 7.1-min delay of the microdialysis sensor was calculated. RESULTS: No delay [-2.2 +/- 6.2 (SD) min] was seen for the needle-type sensor. Instrumental delay was >6.2 min for the microdialysis sensor, accounting for more than 87% of the total reported delay of 7.1 +/- 5.5 min. Mean peak width for the BG curves was 100.8 +/- 25.0 min, for the needle-type sensor curves 110.0 +/- 20.5 min, and for the microdialysis sensor curves 104.6 +/- 21.7 min (P = 0.052 and P = 0.11 vs. BG, respectively). Mean peak width for the uncalibrated microdialysis current values was 105.0 +/- 23.1 min, which was not different from the peak width of the BG curves (P = 0.347). CONCLUSIONS: IG-BG delay may be smaller than previously postulated. The sensor curves tended to have broader peaks than the BG curves, in contrast to the expected narrower peaks predicted by the push-pull phenomenon. This argues against the existence of the push-pull phenomenon. PMID- 17425444 TI - Menstrual cycle effects on insulin sensitivity in women with type 1 diabetes: a pilot study. AB - BACKGROUND: Many women complain of difficulty maintaining euglycemia during the luteal phase of the menstrual cycle. This pilot study's objective was to evaluate possible differences in insulin sensitivity between follicular and luteal phases in women with type 1 diabetes. METHODS: Women using insulin infusion pumps (n = 5, mean age 29.2 +/- 10.9 years, mean body mass index 24 +/- 1.8 kg/m(2)) underwent frequently sampled intravenous glucose tolerance tests during each cycle phase. Insulin sensitivity and glucose effectiveness were determined by Minimal Model analysis. RESULTS: Non-insulin-mediated glucose disposal increased during the luteal phase (0.009 +/- 0.004 min(1)) versus the follicular phase (0.005 +/- 0.003 min(1)) (P < 0.05). Although no significant differences were found in mean insulin sensitivity between follicular (0.76 +/- 0.27 x 10(4)/min(1) /microU/mL) and luteal phase (0.58 +/- 0.26 x 10(4)/min(1) /microU/ mL), three of the five subjects had a decline in insulin sensitivity. CONCLUSIONS: Elevated blood glucose during the luteal phase may increase insulin independent glucose disposal. Some individuals appear more responsive to menstrual cycle effects on insulin sensitivity. Women should be encouraged to use available self-monitoring technology to identify possible cyclical variations in blood glucose that might require clinician review and insulin dosage adjustments. PMID- 17425445 TI - Calibration of the continuous glucose monitoring system for transient glucose monitoring. AB - BACKGROUND: Oral glucose tolerance testing (OGTT) remains the gold standard for diagnosis of diabetes and is used commonly in the research laboratory. The Medtronic MiniMed (North-ridge, CA) Continuous Glucose Monitoring System (CGMS Gold), developed for long-term monitoring of glycemic levels, could provide a convenient means for tracking OGTT results during research protocols; however, the sensor demonstrates glucose and time dependencies that preclude direct application of the company-provided conversion algorithm in the first 12-24 h after sensor insertion. Here, we propose an alternative conversion algorithm that permits utilization of the CGMS monitor for glucose monitoring during this time. METHODS: Healthy female participants underwent CGMS monitoring during OGTT or fasting sessions in combination with finger-stick blood glucose measurements. Logarithmic transformations and multiple regression analysis were used to quantify the time and glucose dependence of the sensors. RESULTS: Sensor performance, as characterized by sensitivity (ratio of sensor current to capillary blood glucose levels), was shown to vary logarithmically with glucose levels as well as time after sensor insertion. A conversion algorithm developed on the basis of these observations was tested on 17 subjects during OGTT. A mean absolute relative difference between capillary blood glucose and CGMS values of 11.6 +/- 6.5% for the new algorithm was seen, compared to 20.6 +/- 5.9% with the Medtronic Solutions version 3.0c algorithm. CONCLUSIONS: Incorporation of the glucose and time dependence in CGMS sensor data yields an improved mean absolute difference between actual and estimated blood glucose values compared to the Medtronic-supplied algorithm in the immediate time period following sensor insertion. PMID- 17425446 TI - Pramlintide as an adjunct to insulin in patients with type 2 diabetes in a clinical practice setting reduced A1C, postprandial glucose excursions, and weight. AB - OBJECTIVE: This study was designed to assess the safety and efficacy of pramlintide therapy in patients with type 2 diabetes in a clinical practice setting. METHODS: In this open-label study, 166 insulin-treated patients with type 2 diabetes added pramlintide therapy (120 microg) during an initiation period in which mealtime insulin was reduced by 30-50%. Insulin doses were subsequently adjusted to optimize glycemic control. Endpoints included safety, as well as change in A1C, postprandial glucose, weight, insulin dose, and patient satisfaction following 6 months of pramlintide treatment. RESULTS: At 6 months, the change in A1C from baseline (8.3%) was -0.56% (P < 0.05; n = 59). Pramlintide treatment significantly reduced mean postprandial glucose excursions (P < 0.05) and weight (-2.8 kg; P < 0.05; n = 125). Glycemic benefits were achieved with lower mealtime insulin doses (-10.3%; P < 0.05; n = 104). Nausea, primarily mild to moderate, was reported by 29.5% of patients (severe nausea in 2.4%). Rates of severe hypoglycemia were low (0.04 events/patient-year). CONCLUSIONS: In this uncontrolled, open-label setting, pramlintide initiation while reducing mealtime insulin, followed by insulin dose optimization, resulted in improvements in postprandial glucose excursions and A1C. These improvements in glycemic control were accompanied by weight loss. PMID- 17425447 TI - Response to Baum et al. on meter miscoding. PMID- 17425448 TI - Glycosylation profiling in clinical proteomics: heading for glycan biomarkers. PMID- 17425452 TI - Electron capture dissociation in the analysis of protein phosphorylation. AB - Protein phosphorylation is a widespread and important post-translational modification. Despite recent advances in phosphoproteomic methods, phosphopeptide identification and site localization remain challenging. Electron capture dissociation has inherent advantages for phosphorylation analysis. The use of electron capture dissociation in this area to date is reviewed and future prospects are outlined. PMID- 17425453 TI - Promise of multiphoton detection in discovery and diagnostic proteomics. AB - Proteomics has lacked adequate methods for handling the complexity (hundreds of thousands of different proteins) and range of protein concentrations (> or =10(6)) of eukaryotic proteomes. New multiphoton-detection methods for ultrasensitive detection of proteins produce 10,000-fold gains in sensitivity and allow highly quantitative, linear detection of 50 zmol (30,000 molecules) to 500 fmol of proteins in complex samples. The potential of multiphoton detection in top-down proteomics analyses is illustrated with applications in monitoring proteomes in very small numbers of cells, in identifying and monitoring complex functional isoforms of cancer-related proteins, and in super-sensitive immunoassays of serum proteins for high-performance detection of cancer. PMID- 17425454 TI - Coupling immunoaffinity techniques with MS for quantitative analysis of low abundance protein biomarkers. AB - The field of proteomics is rapidly turning towards targeted mass spectrometry (MS) methods to quantify putative markers or known proteins of biological interest. Historically, the enzyme-linked immunosorbent assay (ELISA) has been used for targeted protein analysis, but, unfortunately, it is limited by the excessive time required for antibody preparation, as well as concerns over selectivity. Despite the ability of proteomics to deliver increasingly quantitative measurements, owing to limited sensitivity, the leads generated are in the microgram per milliliter range. This stands in stark contrast to ELISA, which is capable of quantifying proteins at low picogram per milliliter levels. To bridge this gap, targeted liquid chromatography (LC) tandem MS (MS/MS) analysis of tryptic peptide surrogates using selected reaction monitoring detection has emerged as a viable option for rapid quantification of target proteins. The precision of this approach has been enhanced by the use of stable isotope-labeled peptide internal standards to compensate for variation in recovery and the influence of differential matrix effects. Unfortunately, the complexity of proteinaceous matrices, such as plasma, limits the usefulness of this approach to quantification in the mid-nanogram per milliliter range (medium abundance proteins). This article reviews the current status of LC/MS/MS using selected reaction monitoring for protein quantification, and specifically considers the use of a single antibody to achieve superior enrichment of either the protein target or the released tryptic peptide. Examples of immunoaffinity assisted LC/MS/MS are reviewed that demonstrate quantitative analysis of low abundance proteins (subnanogram per milliliter range). A strategy based on this technology is proposed for the expedited evaluation of novel protein biomarkers, which relies on the synergy created from the complementary nature of MS and ELISA. PMID- 17425455 TI - Cellular metabolomics of Escherchia coli. AB - Escherichia coli is among the simplest and best-understood free-living organisms. It has served as a valuable model for numerous biological processes, including cellular metabolism. Just as E. coli stood at the front of the genomic revolution, it is playing a leading role in the development of cellular metabolomics: the study of the complete metabolic contents of cells, including their dynamic concentration changes and fluxes. This review briefly describes the essentials of cellular metabolomics and its fundamental differentiation from biomarker metabolomics and lipidomics. Key technologies for metabolite quantitation from E. coli are described, with a focus on those involving mass spectrometry. In particular emphasis is given to the cell handling and sample preparation steps required for collecting data of high biological reliability, such as fast metabolome quenching. Future challenges, both in terms of data collection and application of the data to obtain a comprehensive understanding of metabolic dynamics, are discussed. PMID- 17425456 TI - Proteomic biomarker discovery for the monogenic disease cystic fibrosis. AB - Proteomics was initially viewed as a promising new scientific discipline to study complex disorders such as polygenic, infectious and environment-related diseases. However, the first attempts to understand a monogenic disease such as cystic fibrosis (CF) by proteomics-based approaches have proved quite rewarding. In CF, the impairment of a unique protein, the CF transmembrane conductance regulator, does not completely explain the complex and variable CF clinical phenotype. The great advances in our knowledge about the molecular and cellular consequences of such impairment have not been sufficient to be translated into effective treatments, and CF patients are still dying due to chronic progressive lung dysfunction. The progression of proteomics application in CF will certainly unravel new proteins that could be useful as biomarkers either to elucidate CF basic mechanisms and to better monitor the disease progression, or to promote the development of novel therapeutic strategies against CF. This review will summarize the recent technological advances in proteomics and the first results of its application to address the most important issues in the CF field. PMID- 17425458 TI - Proteomics of Alzheimer's disease: understanding mechanisms and seeking biomarkers. AB - Alzheimer's disease is the scourge of the modern, aging world: a costly, damaging disease that robs the elderly of their ability to function as well as their memories. Three decades of progress have resulted in a deep understanding of the pathological processes and a range of targets for therapy, many of which have advanced to late-stage clinical trials. Proteomics has contributed greatly to these advances and will continue to have a growing role in determining the nature of the pathological lesions in the brain. In addition, proteomics (both gel based and gel free, mass spectrometry based), is likely to play an increasing role in identifying biomarkers that may assist in early diagnosis and in monitoring progression and, most importantly, response to therapy. PMID- 17425459 TI - Approaches to the study of the cell secretome. AB - The secretome, or secretomics, has recently emerged as a new term to describe the global study of proteins that are secreted by a cell, tissue or organism at any given time or under certain conditions. The secretome constitutes an important class of proteins that control and regulate a multitude of biological and physiological processes, thus making it a clinically relevant source for biomarkers and therapeutic target discoveries. There are several approaches that are being implemented to study such a class of proteins; however, each of these approaches has its advantages and limitations. While genome-wide studies using signal predictions can provide a comprehensive analysis of the secretome, the detection and quantification of the actual secreted proteins in a tissue would be more relevant. The goal of this review is to provide an overview of the methods currently used to analyze such a class of proteins, as well as the challenges encountered during the study of the secretome. The implication of studying the cell secretome together with its clinical relevance will be also covered. PMID- 17425457 TI - Mass spectrometry-based strategies for characterization of histones and their post-translational modifications. AB - Due to the intimate interactions between histones and DNA, the characterization of histones has become the focus of great attention. A series of mass spectrometry-based technologies have been dedicated to the characterization and quantitation of different histone forms. This review focuses on the discussion of mass spectrometry-based strategies used for the characterization of histones and their post-translational modifications. PMID- 17425460 TI - Proteomics in atherothrombosis: a future perspective. AB - Atherothrombosis is the primary cause of death in Western countries. The cellular and molecular mechanisms underlying atherosclerosis remain widely unknown. The complex nature of atherosclerotic cardiovascular diseases demands the development of novel technologies that enable discovery of new biomarkers for early disease detection and risk stratification, which may predict clinical outcome. In this review, we outline potential sources and recent proteomic approaches that could be applied in the search of novel biomarkers of cardiovascular risk. In addition, we describe some issues raised in relation to the application of proteomics to blood samples, as well as two novel emerging concepts, such as peptidomics and population proteomics. In the future, the use of high-throughput techniques (proteomic, genomics and metabolomics) will potentially identify novel patterns of biomarkers, which, along with traditional risk factors and imaging techniques, could help to target vulnerable patients and monitor the beneficial effects of pharmacological agents. PMID- 17425461 TI - Autoantibody microarrays for biomarker discovery. AB - Identification of autoantigens and the detection of autoantibody reactivity are useful in biomarker discovery and for explaining the role of important biochemical pathways in disease. Despite all of their potential advantages, the main challenge to working with autoantibodies is their sensitivity. Nevertheless, proteomics may hold the key to overcoming this limitation by providing the means to multiplex. Clearly, the ability to detect multiple autoantigens using a platform such as a high-density antigen microarray would improve sensitivity and specificity of detection for autoantibody profiling. The aims of this review are to: briefly describe the current status of antigen-autoantibody microarrays; provide examples of their use in biomarker discoveries; address current limitations; and provide examples and strategies to facilitate their implementation in the clinical setting. PMID- 17425462 TI - Proteomics in uveal melanoma research: opportunities and challenges in biomarker discovery. AB - Uveal melanoma (UM) is the most frequent primary intraocular tumor in adult humans. Despite the significant advances in diagnosis and treatment of UM in the last decades, the prognosis of UM sufferers is still poor. Metastatic liver disease is the leading cause of death in UM and can develop after a long disease free interval, suggesting the presence of occult micrometastasis. Proteomics technology has opened new opportunities for elucidating the molecular mechanism of complex diseases, such as cancer. This article will review the recent developments in biomarker discovery for UM research by proteomics. In the last few years, the first UM proteomics-based analyses have been launched, yielding promising results. An update on recent developments on this field is presented. PMID- 17425463 TI - Use of nanomaterials in capillary and microchip electrophoresis. AB - This review gives an overview of different separation strategies with nanomaterials and their use in capillary electrophoresis (CE) and capillary electrochromatography, as well as in microchip electrophoresis, including metal and metal oxide nanoparticles, carbon nanotubes, fullerene and polymer nanoparticles, as well as silica nanoparticles. The paper highlights the new developments and innovative applications of nanoparticles as pseudostationary phases or immobilized on the capillary surface for CE separation. The separation and characterization of target nanoparticles with different sizes by CE are reviewed likewise. PMID- 17425464 TI - An array of possibilities in cancer research using cytokine antibody arrays. AB - Protein arrays have shown potential applications in cancer research. After several decades of research, it has become evident that many cytokines are central to the development of cancer and its treatment. Cytokine antibody arrays that have been designed to simultaneously detect multiple cytokines are not only available, but show a diversity of applications in the study of many diseases in addition to cancer. This review will focus on the implementation of cytokine antibody arrays in many aspects of cancer research, such as biomarker discovery, molecular mechanisms of cancer development, preclinical studies and the effects of cancer compounds. PMID- 17425465 TI - Proteomic approach to aging research. AB - The scope of the current paper is to review existing and potential applications of proteomic analysis to aging research. The focus will lie on the unique opportunities of high-throughput studies for uncovering specific alterations in protein expression, protein complexes or protein modifications caused by biological aging. The result of such studies will outline aging phenotypes and potentially indicate pathways involved in the pathogenesis of age-associated disfunctions. Specific attention is paid to the illustrations of successful applications of proteomic technologies and potential applications of new proteomic concepts to biogerontological studies. PMID- 17425467 TI - Genetic engineering for skeletal regenerative medicine. AB - The clinical challenges of skeletal regenerative medicine have motivated significant advances in cellular and tissue engineering in recent years. In particular, advances in molecular biology have provided the tools necessary for the design of gene-based strategies for skeletal tissue repair. Consequently, genetic engineering has emerged as a promising method to address the need for sustained and robust cellular differentiation and extracellular matrix production. As a result, gene therapy has been established as a conventional approach to enhance cellular activities for skeletal tissue repair. Recent literature clearly demonstrates that genetic engineering is a principal factor in constructing effective methods for tissue engineering approaches to bone, cartilage, and connective tissue regeneration. This review highlights this literature, including advances in the development of efficacious gene carriers, novel cell sources, successful delivery strategies, and optimal target genes. The current status of the field and the challenges impeding the clinical realization of these approaches are also discussed. PMID- 17425468 TI - Acoustic detection of coronary artery disease. AB - Coronary artery disease (CAD) occurs when the arteries to the heart (the coronary arteries) become blocked by deposition of plaque, depriving the heart of oxygen bearing blood. This disease is arguably the most important fatal disease in industrialized countries, causing one-third to one-half of all deaths in persons between the ages of 35 and 64 in the United States. Despite the fact that early detection of CAD allows for successful and cost-effective treatment of the disease, only 20% of CAD cases are diagnosed prior to a heart attack. The development of a definitive, noninvasive test for detection of coronary blockages is one of the holy grails of diagnostic cardiology. One promising approach to detecting coronary blockages noninvasively is based on identifying acoustic signatures generated by turbulent blood flow through partially occluded coronary arteries. In fact, no other approach to the detection of CAD promises to be as inexpensive, simple to perform, and risk free as the acoustic-based approach. Although sounds associated with partially blocked arteries are easy to identify in more superficial vessels such as the carotids, sounds from coronary arteries are very faint and surrounded by noise such as the very loud valve sounds. To detect these very weak signals requires sophisticated signal processing techniques. This review describes the work that has been done in this area since the 1980s and discusses future directions that may fulfill the promise of the acoustic approach to detecting coronary artery disease. PMID- 17425469 TI - Glucocorticoid replacement therapy in patients with Addison's disease. AB - One hundred and fifty years ago, Thomas Addison published his classic paper on the 'Constitutional and Local Effects of Disease of the Supra-renal Capsules', in which he described 11 patients with the disorder that would come to bear his name. Adrenal insufficiency is a rare disease, but its prevalence is increasing. The most frequent causes of adrenal insufficiency in western countries are autoimmune adrenalitis, but other causes include, tuberculosis systemic fungal infections, AIDS, metastatic carcinoma and isolated glucocorticoid deficiency. It is clear that autoimmunity precedes overt Addison's disease by years, as in many autoimmune endocrine disorders. Adrenocortical function is lost over a period of years as it progresses to overt Addison's disease. This editorial discusses the controversial glucocorticoid replacement therapy in patients with Addison's disease, and aims to provide a good review of literature and suggested guidelines for appropriate treatment of this disease. PMID- 17425470 TI - Recommendations for the management of migraine in paediatric patients. AB - Migraine is a common and disabling condition in children and adolescents. The complexity of migraine on a pathogenetic and clinical level results from the interaction between biological, psychological and environmental factors. Appropriate management requires an individually tailored strategy giving due consideration to both pharmacological and non-pharmacological measures. Ibuprofen (7.5-10.0 mg/kg) and acetaminophen (15 mg/kg) are safe and effective, and should be considered for symptomatic treatment. Sumatriptan nasal spray (5 and 20 mg) is also likely to be effective, but at the moment, should be considered for the treatment of adolescents only. With reference to prophylactic drug treatment, the available data suggest that flunarizine (5 mg/day) is likely to be effective and pizotifen and clonidine are likely to be ineffective. The efficacy data regarding propranolol, nimodipine and trazodone are conflicting. Insufficient evidence is available on cyproheptadine, amitriptyline, divalproex sodium, topiramate, levetiracetam, gabapentin or zonisamide. The management of migraine in children needs an individualised therapeutic approach, directed to the whole person of the child, taking into account the developmental perspective and the high rate of psychiatric comorbidities. It is the authors' opinion that for the prophylaxis of migraine, interventions such as identification and avoidance of trigger factors, regulation of lifestyle, relaxation, biofeedback, cognitive behavioural treatment and psychological or psychotherapeutic interventions (e.g., psychodynamics) could be much more effective than pharmacotherapy. PMID- 17425471 TI - Review of long-acting stimulants in the treatment of attention deficit hyperactivity disorder. AB - A number of long-acting medications for the treatment of attention deficit hyperactivity disorder (ADHD) have recently been developed and approved for use in the US. These compounds are intended to optimize and maintain ADHD symptom control throughout the day, while eliminating problems associated with short acting medications, such as the need for in-school, midday or multiple daily doses. Recent reports confirm that the safety and tolerability of long-acting medications are similar to those of short-acting medications, although long acting medications appear to have a lower risk of abuse and diversion and may be associated with significant improvements in medication adherence. Distinctions can be made among the long-acting medications with regard to the onset, magnitude and duration of their clinical effects. Recognition of these differences is important for individualizing treatment for patients with ADHD. PMID- 17425472 TI - Pharmacological antithrombotic adjuncts to percutaneous coronary intervention. AB - Stent thrombosis is the major cause of early adverse events during percutaneous coronary intervention. Its incidence has fallen considerably in recent years, principally due to the introduction of effective antithrombotic therapies. The selection of an appropriate antithrombotic regimen is critical in achieving a balance between reducing ischaemic events and minimising bleeding complications in patients undergoing percutaneous coronary intervention. In this article, evidence for the role of antiplatelet and anticoagulant therapies is discussed, including the thienopyridines, glycoprotein IIb/IIIa receptor antagonists, direct thrombin inhibitors and pentasaccharides. PMID- 17425473 TI - Inhaled therapy for stable chronic obstructive pulmonary disease. AB - Bronchodilators (anticholinergic drugs and beta2-agonist drugs) represent the mainstay of chronic obstructive pulmonary disease treatment and can be combined with inhaled corticosteroids for greater efficacy. This review examines the role of short- and long-acting cholinergic drugs, together with that of short- and long-acting beta2-agonist drugs. A brief analysis of the long-term use of corticosteroids in chronic obstructive pulmonary disease is made, with a review of the safety and possible side effects. Combination therapy is more frequently used due to its practicality and capacity to enhance compliance. The main studies on treatment combinations are also analyzed in this paper. PMID- 17425474 TI - Adult growth hormone deficiency: to treat or not to treat. AB - In the last 15 years, it has been recognised that growth hormone deficiency (GHD) in the adult leads to increased morbidity (metabolic syndrome, osteoporosis, muscle wasting, impaired quality of life) and increased incidence of cardiovascular events, a main cause of the increased mortality observed in this population. Pituitary adenomas and their treatment (surgery, radiation) are the most common cause of GHD in adults. Patients with biochemical diagnosis of severe GHD must be offered growth hormone replacement therapy only in the presence of GHD associated morbidity. This treatment improves morbidity, but its effect on mortality remains to be proven. Continuation of treatment for GHD patients in late adolescence to early adulthood, who have reached final height with growth hormone replacement, requires careful clinical judgement. PMID- 17425475 TI - Chemotherapy for advanced gastric cancer: across the years for a standard of care. AB - Chemotherapy is of crucial importance in advanced gastric cancer patients, in order to obtain palliation of symptoms and improve survival. The most extensively studied drugs as single agents are 5-fluorouracil, cisplatin, doxorubicin, epirubicin, mitomycin C and etoposide. Newer chemotherapeutic agents include the taxanes (docetaxel and paclitaxel), oral fluoropyrimidines (capecitabine and S 1), oxaliplatin and irinotecan. Randomised trials comparing monotherapy with combination regimens have consistently shown increased response rates in favour of combination regimens, whereas only marginally improved survival rates were usually found. Several combination therapies have been developed and have been examined in Phase III trials. However, in most cases, they have failed to demonstrate a survival advantage over the reference arm. There is no internationally accepted standard of care, and uncertainty remains regarding the choice of the optimal chemotherapy regimen. The objective of this article is to review the present literature available on major Phase II - III clinical trials, in which patients suffering from advanced gastric cancer were treated with cytotoxic chemotherapy. PMID- 17425476 TI - Uterine papillary serous carcinoma: a review. AB - The purpose of this article is to review the available literature for uterine papillary serous carcinoma (UPSC). A literature search was conducted to identify publications on UPSC. The literature on UPSC is composed mainly of retrospective, single-institution reports. Despite these limitations, several recommendations can be made. When UPSC is confirmed on preoperative biopsy, complete surgical staging should be performed. Although whole abdominal radiotherapy has a limited role in early-stage UPSC, there may be a role for postoperative chemotherapy in early-stage UPSC. In the setting of optimally debulked advanced-stage disease, a combination of radiation and chemotherapy may be indicated. In the setting of recurrent or suboptimally debulked advanced disease, a platinum-based regimen is recommended. Although comprising a minority of the women with endometrial cancer, women with UPSC do account for a disproportionate percentage of the recurrences. There is a need for clinical trials to determine the optimal therapy for this cohort of patients. PMID- 17425477 TI - Rivastigmine and Parkinson dementia complex. AB - Patients suffering from Parkinson's disease dementia (PDD) have a movement disorder, but it can be difficult to determine whether the functional impairment, which is critical in making the assessment of whether a patient has achieved the threshold for a diagnosis of dementia, is due to the dementia or the underlying Parkinson's disease. Although the cognitive impairment found in nondemented patients with Parkinson's disease is very dysexecutive in nature, the DSM IV (Diagnostic and Statistical Manual of Mental Disorders of the American Psychiatric Association IV) diagnosis of PDD has memory impairment as the defining characteristic of PDD. Severe deficits in cortical, cholinergic, excitatory, neuromodulatory input mean that memory impairment is not always due to encoding and retrieval strategy deficits, but it may also be amnesic without being related to concomitant Alzheimer's disease pathology. Patients with PDD have a high mortality, especially when they develop hallucinations and/or are admitted to nursing homes. Of interest is the reduction in mortality that was more marked in the subgroup with visual hallucinations at baseline. The increased mortality in PD may be due to autonomic failure, evidenced by the reductions in heart rate variability in these patients. This reduction is greater in patients with hallucinations. Rivastigmine is a dual inhibitor of brain acetyl- and butyrylcholinesterases that has been evaluated in the symptomatic treatment of patients with mild-to-moderate dementia associated with idiopathic Parkinson's disease. Although there is a need for more studies using pragmatic measures, such as time to residential care facility and both patient and carer quality of life assessments, rivastigmine appears to improve cognition and activities of daily living in patients with PDD, resulting in a clinically meaningful benefit in a large number of cases. PMID- 17425478 TI - Adalimumab for the treatment of ankylosing spondylitis. AB - Ankylosing spondylitis is a chronic inflammatory disease, with a prevalence of approximately 0.5%, which starts in the third decade of life. Treatment was, until recently, limited. Conventional disease-modifying drugs are not effective for the spinal manifestations, and NSAIDs and physical therapy were the standard treatment, without any other options for patients who did not respond to this treatment. Therefore, the high efficacy of the new group of TNF-blockers for the treatment of active ankylosing spondylitis represents a breakthrough for NSAID refractory patients. Following the introduction of the two TNF-blockers, infliximab and etanercept, the fully humanized, anti-TNF monoclonal antibody adalimumab is now the third product that has been approved for the treatment of ankylosing spondylitis. Adalimumab is given subcutaneously every 2 weeks at a dose of 40 mg. In open and placebo-controlled trials, the drug was shown to be safe and effective in ankylosing spondylitis patients. Long-term treatment data of up to 2 years are now available, confirming efficacy and acceptable safety. PMID- 17425479 TI - Tipranavir: a new protease inhibitor for the treatment of antiretroviral experienced HIV-infected patients. AB - Tipranavir (TPV) is a novel non-peptidic protease inhibitor (PI). It binds strongly and selectively to the HIV-1 protease, is orally administered twice daily, boosted with low doses of ritonavir, and shows a favourable resistance profile. In the two registrational trials, named RESIST 1 and 2, TPV/ritonavir 500/200 mg b.i.d., along with an optimised antiretroviral backbone, provided better virologic responses than controls receiving standard of care ritonavir boosted PI-based regimens. A total of 21 mutations at 16 protease codons have been shown to impact on TPV susceptibility and response rates. The TPV mutation score includes L10V, I13V, K20M/R/V, L33F, E35G, M36I, K43T, M46L, I47V, I54A/M/V, Q58E, H69K, T74P, V82L/T, N83D and I84V. Viruses containing eight or more of these mutations are generally resistant to the drug. TPV use is associated with an excess of grade 3/4 liver enzyme elevations compared with other ritonavir-boosted PIs, and the potential for drug-drug interactions is relevant and must be considered when prescribing TPV. PMID- 17425480 TI - Efavirenz: a review. AB - Efavirenz is a non-nucleoside reverse transcriptase inhibitor that in most treatment guidelines is recommended to be taken combined with two nucleoside analogue reverse transcriptase inhibitors, as a preferred first-line regimen for the treatment of HIV-1 infection. The antiretroviral efficacy of efavirenz-based combination regimens is good, as has been demonstrated in many clinical trials. Efavirenz has a long plasma half-life, which allows for once-daily dosing, but, as a consequence of this and the low genetic barrier, it is also prone to select for viral resistance when adherence to therapy is suboptimal. The most frequently encountered side effects are neuropsychiatric symptoms. These side effects are usually transient, but have been shown to persist for up to 2 years after initiation of therapy in some patients. This review outlines important and recent pharmacological and clinical data, which explain why efavirenz became a component of preferred treatment regimens today. PMID- 17425481 TI - LJP-394 (abetimus sodium) in the treatment of systemic lupus erythematosus. AB - Renal disease is one of the most severe aspects of systemic lupus erythematosus (SLE), potentially leading to irreversible kidney failure. The standard of care for severe lupus nephritis involves the use of high-dose corticosteroids, cyclophosphamide and other immunosuppressive drugs. Although these drugs are effective in controlling disease activity in the majority of patients, up to 25% of patients treated with cyclophosphamide-based protocols develop renal insufficiency and end-stage renal disease, and treatment discontinuation is associated with the occurrence of flares. Furthermore, these therapies are associated with a high incidence of short- and long-term side effects. LJP-394 (abetimus sodium) is an investigational agent specifically designed to decrease anti-dsDNA antibody levels, and it is under development for the prevention of nephritic flares in patients with SLE since the early 1990s. The drug has been evaluated in 13 clinical trials that evaluated > 800 patients with SLE, over a 10 year time span. It is likely that LJP-394 might have a role in the prevention of renal flares in SLE patients, and if the initial data is confirmed in an ongoing trial, this drug could represent either a substitute for immunosuppressive drugs or could allow a reduction of their dose, thereby reducing the risks of short- and long-term side effects. This paper reviews the principal aspects of chemistry, pharmacokinetics, efficacy and safety of LJP-394, and analyses studies on animal models and clinical studies conducted in the last few years. PMID- 17425482 TI - Clinical trials update from the World Congress of Cardiology 2006. AB - This article provides information on trials presented at the World Congress of Cardiology 2006 (WCC-2006). This was a joint meeting of the European Society of Cardiology and the World Heart Federation. Heart failure trials dealing with understudied patient populations (e.g., the elderly), trials of new treatments (i.e., immune modulation therapy) and new intervention strategies (i.e., the Cardiac Insufficiency Bisoprol-III study) or patient monitoring (the Home or Hospital Heart Failure study) were presented at this meeting. Among the studies about coronary artery disease treatment, WCC-2006 saw the presentation of the landmark meta-analyses regarding the incidence of late events in patients receiving drug-eluting stents. The main results of these meta-analyses, and the associated debate, are summarised in this article. Finally, the results of other trials of percutaneous patent foramen ovale closure and atrial fibrillation treatment in elderly patients are summarised. PMID- 17425483 TI - Place of electroencephalograpic biofeedback for attention-deficit/hyperactivity disorder. PMID- 17425484 TI - Viruses in the treatment of malignant glioma. PMID- 17425485 TI - New concept of stroke pathophysiology: the potential impact on stroke management. PMID- 17425487 TI - Desmoteplase in the treatment of acute ischemic stroke. AB - Desmoteplase, developed by Paion, Forest and Lundbeck, is a novel plasminogen activator that selectively activates fibrin-bound plasminogen and is currently being investigated for the treatment of acute ischemic stroke within the time window of 3-9 h after symptom onset. Desmoteplase is believed to offer pharmacologic advantages over currently approved treatment options. To date, two published Phase II perfusion imaging-based clinical trials have reported the safety and potential efficacy of desmoteplase in ischemic stroke. Results from a recently completed Phase III trial in Europe, Asia and the USA are awaited. This article reviews the available data on desmoteplase, including discussion of its favorable features and potential benefit beyond the 3-h time window in the treatment of ischemic stroke. PMID- 17425489 TI - Psychiatric manifestations of brain tumors: diagnostic implications. AB - Brain tumors can cause any type of psychiatric symptoms. Rarely, brain tumors can present without any localizing signs but with psychiatric symptoms. A review of the literature indicates that there is no association between psychiatric symptoms and tumor location or histological type. Hence, it is important for clinicians to have an index of suspicion of brain tumor in patients with new onset psychiatric symptoms, atypical presentations and treatment resistance and, as a result, consider neuroimaging. Early detection is of paramount importance for treatment and quality of life of patients. Mood symptoms may herald a brain tumor. For example, anorexia can be a presenting symptom of hypothalamic tumors. A prospective controlled study will be of benefit to further assess the association between psychiatric symptoms and brain tumor locations. New developments in the diagnostic and treatment area of brain tumors would be of help in undertaking such a prospective study. PMID- 17425490 TI - Targeting the dopamine system in the treatment of attention-deficit/hyperactivity disorder. AB - Attention-deficit/hyperactivity disorder (ADHD) is a highly heritable condition that affects a significant number of children and adults worldwide. During the past 30 years, the diagnosis and treatment of ADHD has relied on clinical assessment and empirical experience with stimulant medications. More recently, advances in family genetic studies, molecular genetic studies, preclinical research, radiographic imaging techniques and neuropsychological evaluation have significantly enhanced our understanding of the neurobiology of ADHD. This review highlights the current central role of dopamine in the pathophysiology and treatment of ADHD and implications for future advances in diagnosis and treatment. PMID- 17425491 TI - Use of nanoparticles for drug delivery in glioblastoma multiforme. AB - Treatment of glioblastoma multiforme (GBM), a primary malignant tumor of the brain, is one of the most challenging problems as no currently available treatment is curative. Surgery remains the basic treatment in which the bulk of the tumor is removed and the peripheral infiltrating part is the target of supplementary treatments. The currently available anticancer therapeutics have less than optimal usefulness for GBM, mainly owing to delivery problems to the tumor, including those due to blood-brain barrier. Several new therapies in development for GBM will require innovative methods of delivery besides the current practice of direct introduction of drugs and devices into the tumor. Nanobiotechnology, particularly nanoparticles, is making a significant contribution to the improvement of drug delivery in cancer and many of these technologies can be applied to GBM. Nanobiotechnology can provide a platform for combination of diagnostics, therapeutics, and its delivery to the tumor with subsequent monitoring of response. These technologies are reviewed and the prospects of improvement of therapeutic delivery to the brain are excellent during the next 5 years. Curing of GBM, however, depends on discovery of an anticancer agent that will destroy GBM completely. PMID- 17425492 TI - What is the best adjuvant treatment for very young patients with medulloblastoma? AB - The standard treatment for medulloblastoma is surgery followed by adjuvant chemotherapy and external beam radiotherapy to the craniospinal axis and posterior fossa. However, in very young children, craniospinal irradiation has a more significant detrimental effect in terms of neurocognitive function and growth. This article reviews the different strategies used for very young patients with medulloblastoma. PMID- 17425493 TI - Treatment of arterial ischemic stroke in children. AB - Stroke is a rare but increasingly recognized disorder in children. Current therapies for arterial ischemic stroke include thrombolytic, antithrombotic and antiplatelet agents, blood transfusion and surgery. Adult studies, pediatric case studies and expert opinion form the basis for these treatment strategies. Thrombolytic agents are increasingly used but, as in adults, the majority of arterial ischemic strokes in children are treated with antiplatelet and antithrombotic agents. Sickle-cell patients, a distinct subset of the pediatric stroke population, are treated primarily with transfusion therapy. Pediatric arterial ischemic stroke studies are needed to determine the most appropriate course of treatment. An international study is currently in progress to formally study the incidence, risk factors, treatment strategies and outcomes of stroke in children. PMID- 17425494 TI - Functional neuroimaging in post-traumatic stress disorder. AB - Traumatic stress has a broad range of effects on brain function. Brain areas implicated in the stress response include the amygdala, hippocampus, and prefrontal cortex. Brain studies in patients with post-traumatic stress disorder replicated findings in animal studies by finding alterations in these brain areas. Brain areas implicated in post-traumatic stress disorder play an important role in the stress response as well as memory, highlighting the important interplay between memory and the traumatic stress response. Future studies are required to assess the relationship between recovery from traumatic stress and changes in brain function. PMID- 17425496 TI - Neuroleptic malignant syndrome in elderly patients. AB - Antipsychotic drugs are widely and increasingly prescribed for neurobehavioral disorders in elderly patients. However, the efficacy of these drugs has not been consistently demonstrated in geriatric populations and there are continuing concerns regarding adverse effects. Among the latter are severe neurological disorders, including neuroleptic malignant syndrome. Although the incidence and mortality of neuroleptic malignant syndrome may have declined with heightened awareness of this disorder and the development of newer drugs, neuroleptic malignant syndrome still occurs in association with the use of antipsychotics. To enhance patient safety and clinical vigilance among practitioners, the authors present a clinical overview of neuroleptic malignant syndrome. PMID- 17425495 TI - Dementia syndromes: evaluation and treatment. AB - As our population ages, diseases affecting memory and daily functioning will affect an increasing number of individuals, their families and the healthcare system. The social, financial and economic impacts will be profound. This article provides an overview of current dementia syndromes to assist clinicians in evaluating, educating and treating these patients. PMID- 17425498 TI - Microspheres of collagen-apatite nanocomposites with osteogenic potential for tissue engineering. AB - Microparticulate systems have attracted a great deal of attention over the past few years as a carrier for the delivery of cells and proteins in the treatment of defective tissues. The composition of microparticulates is regarded as being of utmost importance for the successful recruitment of the cells involved in the tissue regeneration process. Collagen-apatite nanocomposites mimicking the extracellular bone matrix are thus considered to be a potential vector for bone regeneration, either directly or through the delivery of osteogenic cells. In this study, we developed microspheres constituted of collagen and apatite for the treatment of skeletal defects. The apatite-precipitated collagen solution (30% apatite) was formed into microspheres under a water-in-oil emulsion condition. Spherical particles with diameters of tens to hundreds of micrometers (average of approximately 166 microm) were successfully produced. The internal structure of the microspheres featured a typical nanocomposite wherein apatite nanocrystalline precipitates were organized evenly within the reconstituted collagen matrix. The nanocomposite microspheres were observed to recruit favorable adhesion and growth of rat bone marrow derived stem cells. The cells supported on the nanocomposite microspheres stimulated the expression of a series of bone-associated genes. The osteogenic marker, alkaline phosphatase, was secreted to a significantly higher level on the nanocomposite microspheres than on the pure collagen counterpart. The present finding suggests that the collagen-apatite nanocomposite microspheres have high osteogenic potential and are useful for tissue-engineering applications. PMID- 17425500 TI - Recent progress in dendrimer-based nanocarriers. AB - A large number of drug delivery systems--mostly in the form of liposomes, microspheres, nanoparticles and hydrogels--have been designed to achieve targeted delivery and sustained release of drugs by exploiting the inherent properties of polymers. The size, shape, and surface properties of the polymer are used to modulate the pharmacokinetic and pharmacodynamic behavior of drugs conjugated with or encapsulated in the polymeric carrier. Recently, a class of well-defined, monodisperse, and tree-like polymers called dendrimers has attracted attention because of the flexibility they offer in terms of their size, shape, branching, length, and surface functionality. A unique characteristic of dendrimers is that they can act as a particulate system while retaining the properties of a polymer. Drugs and diagnostic agents can be encapsulated in the central core or bound to the surface of the dendrimer by noncovalent or covalent interaction. Dendritic polymers can significantly improve pharmacokinetic and pharmacodynamic properties of low molecular weight and protein-based therapeutic agents. Furthermore, fluorescent antibodies and imaging contrast agents can be bound to these new polymers and the resulting complexes can be used for analyzing biological fluids and for diagnosis. Because of their size, shape, and ability to conjugate with a wide range of chemical entities, dendrimers have found many applications in the pharmaceutical and biomedical sciences. This review focuses on the unique carrier properties of biomimetic dendrimers and discusses a wide range of applications of dendrimers in drug delivery, including their use as drug solubilizers, absorption enhancers, release modifiers, and carriers for targeting drugs and diagnostic agents. PMID- 17425501 TI - Lipoproteins: from physiological roles to drug delivery potentials. AB - Lipoproteins, the endogenous lipid-protein associations responsible for lipid metabolism within the human body, have attracted interest in recent years for their potential as drug delivery carriers owing to, mainly, their lipophilic/amphiphilic nature, which makes them ideal for interacting with highly lipophilic drugs. After lipoprotein particles have been isolated from the blood, drugs can be "loaded" onto them with a variety of methods. Loading can be done either in soluble/suspended form in a liquid medium or as a dry film. Each method has advantages and disadvantages. The drug-loaded lipoproteins can be modified by the attachment of different ligands that target the particles to specific tissue/cell types within the body. A wide variety of drug molecules both from small molecular or macromolecular structures have been tested successfully, mostly in vitro, for their potential for delivery by lipoprotein carriers. PMID- 17425502 TI - Stem cell marker OCT3/4 in tumor biology and germ cell tumor diagnostics: history and future. AB - The transcription factor OCT3/4 (also known as POU5F1 and Oct4) is regarded as one of the key regulators of pluripotency. Expression in nonmalignant cells is restricted to the pluripotent cells in the embryo and the primordial germ cells that will pass pluripotency to future generations via the gametes. Although major progress has been made in successfully identifying other players in the pluripotency network by using high-throughput screening methods, the exact mechanisms involved in regulation of OCT3/4 in vivo remain largely to be elucidated. In human tumors, OCT3/4 is the most informative marker in germ cell tumors diagnostics and is expressed in the precursor lesions gonadoblastoma and carcinoma in situ, as well as in invasive embryonal carcinoma and seminomatous tumors. Currently, the application of OCT3/4 for screening in high-risk patient populations is the novel focus of study. This article reviews OCT3/4 expression in normal development and germ cell tumors. In addition, expression in adult tissues and nongerm cell tumor malignancies in relation to splice variants and pseudogenes is discussed. An overview of the upstream and downstream factors in the OCT3/4 pathway as well as the epigenetic regulation of the gene is summarized and the possible role in oncogenesis considered. PMID- 17425503 TI - Emerging technologies for genome-wide DNA methylation profiling in cancer. AB - Genome-wide epigenetics, namely, epigenomics, is one of the current frontiers in cancer genomics. Epigenetic silencing of tumor-suppressor genes plays an important role in the pathogenesis of most cancers. Several technologies are emerging for the genome-wide profiling of DNA methylation to find new tumor suppressor genes and predictions for clinical phenotypes in individual patients. PMID- 17425504 TI - Downregulation of gap junctions in cancer cells. AB - Gap junctions are intercellular plasma membrane domains enriched in channels that allow direct exchange of ions and small molecules between adjacent cells. Gap junction channels are composed of a family of transmembrane proteins called connexin. Connexins play important roles in the regulation of cell growth and differentiation. Cancer cells usually have downregulated levels of gap junctions, and several lines of evidence suggest that loss of gap junctional intercellular communication is an important step in carcinogenesis. In support of this hypothesis are studies showing that reexpression of connexins in cancer cells causes normalization of cell growth control and reduced tumor growth. To gain a more detailed understanding of the role of connexins as tumor suppressors, a clearer picture of the mechanisms involved in loss of gap junctions in cancer cells is needed. Furthermore, defining the mechanisms involved in downregulation of connexins in carcinogenesis will be an important step toward utilizing the potential of connexins as targets in cancer prevention and therapy. Various mechanisms are involved in the loss of gap junctions in cancer cells, ranging from loss of connexin gene transcription to aberrant trafficking of connexin proteins. This review will discuss our current knowledge on the molecular mechanisms involved in the downregulation of gap junctions in cancer cells. PMID- 17425505 TI - Recurrent fusion oncogenes in carcinomas. AB - Chromosome structural aberrations giving rise to fusion oncogenes is one of the most common mechanisms in oncogenesis. Although this type of gene rearrangement has long been recognized as a fundamental pathogenetic mechanism in hematologi cal malignancies and soft-tissue tumors, it has until recently only rarely been described in the common carcinomas. In this review, the existing information on recurrent fusion oncogenes characterizing carcinomas is summarized, namely, the RET and NTRK1 fusion oncogenes in papillary thyroid carcinoma, PAX8-PPARG in follicular thyroid carcinoma, MECT1-MAML2 in mucoepidermoid carcinoma, the TFE3 and TFEB fusion oncogenes in kidney carcinomas, BRD4-NUT in midline carcinomas, ETV6-NTRK3 in secretory breast carcinomas, and TMPRSS2-ETS fusion oncogenes in prostate carcinomas. As in hematological and soft-tissue malignancies, the most common types of genes involved in fusion oncogenes in carcinomas are transcription factors and tyrosine kinases. With a few exceptions, most fusion oncogenes are tumor type specific in carcinomas, as in other cancers. The mechanisms behind the relative specificity of this type of somatic mutation involve the cellular environment influencing the selection of oncogenic fusions, and the oncogenic fusions in turn driving differentiation programs that may alter the cellular environment. The data summarized on different types of carcinomas characterized by fusion oncogenes indicate that the pathogenetic mechanisms involved in epithelial carcino-genesis may be similar to those known to operate in hematological and soft-tissue malignancies, and further anticipates that many more fusion oncogenes await identification in the most common types of human cancer. PMID- 17425506 TI - Colorectal cancer: a multipathway disease. AB - The linear sequence of genetic alterations illustrated in the Vogelstein model provides a readily understandable illustration of the fundamental principles underlying colorectal tumorigenesis. However, it is now clear that colorectal cancer is a multi-pathway disease. In this review, the concept that inactivation of the tumor suppressor gene APC serves to initiate virtually all colorectal cancers is shown to be an oversimplification. APC inactivation may have important tumorigenic pathogenic effects beyond the mere initiation of precancerous adenomas. Furthermore, the early evolution of colorectal neoplasia must sometimes occur by mechanisms other than inactivation of APC or related alterations that would drive dysregulated Wnt pathway signaling. Oncogenic mutations implicating both BRAF and KRAS are highlighted as alternative initiating steps that synergize with DNA methylation and occur within the context of serrated polyps. CRC comprises subgroups with particular clinical, pathological, and molecular features. PMID- 17425507 TI - Novel animal models to study the role of the growth hormone insulin-like growth factor I axis in prostate cancer. PMID- 17425508 TI - Hrs makes receptors silent: a key to endosomal protein sorting. PMID- 17425509 TI - Normal and malignant germ cell development. PMID- 17425510 TI - The function of ubiquitin-binding protein complexes in endocytic downregulation of growth factor receptors. PMID- 17425511 TI - Diagnostic and prognostic value of molecular cytogenetic analyses of sextant biopsies from prostate cancer suspects. PMID- 17425512 TI - Azo chemistry and its potential for colonic delivery. AB - Improved delivery systems are needed for drugs currently in use to treat localized diseases of the colon. One promising approach is to deliver the drugs specifically to the colon, an approach that has gained importance recently in the treatment of these diseases. The advantages of targeting drugs specifically to the diseased colon include fewer systemic side effects, a need for lower doses of drugs, and maintenance of the drug in its intact form close to the target site. The potential for colon-specific delivery of therapeutic proteins and peptides is also of interest. To achieve colon-specific drug delivery following oral administration, the drug needs to be protected from absorption by the upper gastrointestinal tract and from degradation by the upper gastrointestinal tract environment, allowing the drug to be abruptly released into the proximal colon. One strategy for targeting orally administered drugs to the colon exploits carriers that are degraded specifically by colonic bacteria and utilizes microbially degradable polymers/drugs, especially azo-cross-linked polymers/drugs. Prodrugs utilizing azo linkages are sulfasalazine, ipsalazine, balsalazine, and olsalazine. These were developed for delivery of 5-amino salicylic acid to the colon for localized chemotherapy of inflammatory bowl disease. The azo-conjugation approach utilizes the ability of the colonic environment to cleave these conjugates and protects the drug from absorption or degradation in the upper gastrointestinal tract. It is believed that flavin mediators present in the colon and azo-reductase enzymes released from colonic bacteria are responsible for the degradation of azo-aromatic compounds for site specific delivery of the drug to the colon. PMID- 17425513 TI - Antibodies and peptides in cancer therapy. AB - Antibodies and peptides play a variety of roles in cancer therapy: monoclonal antibodies (mAbs) and peptides are directly used in anticancer therapy and also as targeting moieties. More than 10 mAbs, either unconjugated or in immunoconjugate forms have been approved for use in oncology. Various peptides, tumor necrosis factors (TNFs), and cell surface-targeted peptides have been identified using phage-display libraries. In combination with standard chemotherapeutic agents, mAbs and peptides can significantly increase the toxicity of anticancer drugs to tumors and reduce their adverse side effects on normal tissues. The present review summarizes some major applications of antibodies and peptides in cancer therapy. PMID- 17425514 TI - Salt intake and insulin sensitivity in healthy human volunteers. AB - The literature on salt intake and insulin sensitivity presents a mixed picture, as some studies have shown an increase, whereas others have shown a decrease, in insulin action as sodium intake is enhanced. In some cases, this may relate to the study of salt intake in patients with co-morbidities such as hypertension or diabetes. In the present study, we selected healthy normotensive lean volunteers who underwent a euglycaemic clamp following 6 days of a low-salt diet (20 mmol sodium daily) and, subsequently, 6 days of a high-salt diet (200 mmol sodium daily). Our results show an increase in insulin-mediated glucose disposal during euglycaemic clamp conditions that was significantly higher following the high salt diet compared with the low-salt diet (7.41+/-0.41 compared with 6.11+/-0.40 mg x kg(-1) of body weight x min(-1) respectively; P=0.03). We measured calf blood flow before and during insulin infusion (no significant change after the two dietary salt interventions was detected) and plasma non-esterified fatty acids (also no significant differences were detected). We observed the expected increases in renin concentration and aldosterone activity in subjects on the low salt diet, and also observed a significantly less increase in plasma noradrenaline concentration during euglycaemic insulin infusion following the high-salt compared with the low-salt diet. We propose that the 4-5-fold increase in serum aldosterone and the greater increase in plasma noradrenaline concentration following the low-salt intervention compared with the high-salt period may have contributed to the differences in insulin sensitivity following the adjustment in dietary sodium intake. PMID- 17425515 TI - A reversible form of lysine acetylation in the ER and Golgi lumen controls the molecular stabilization of BACE1. AB - The lipid second messenger ceramide regulates the rate of beta cleavage of the Alzheimer's disease APP (amyloid precursor protein) by affecting the molecular stability of the beta secretase BACE1 (beta-site APP cleaving enzyme 1). Such an event is stimulated in the brain by the normal process of aging, and is under the control of the general aging programme mediated by the insulin-like growth factor 1 receptor. In the present study we report that BACE1 is acetylated on seven lysine residues of the N-terminal portion of the nascent protein. This process involves lysine acetylation in the lumen of the ER (endoplasmic reticulum) and is followed by deacetylation in the lumen of the Golgi apparatus, once the protein is fully mature. We also show that specific enzymatic activities acetylate (in the ER) and deacetylate (in the Golgi apparatus) the lysine residues. This process requires carrier-mediated translocation of acetyl-CoA into the ER lumen and is stimulated by ceramide. Site-directed mutagenesis indicates that lysine acetylation is necessary for nascent BACE1 to leave the ER and move ahead in the secretory pathway, and for the molecular stabilization of the protein. PMID- 17425516 TI - Non-hypertensive tetraPEGylated canine haemoglobin: correlation between PEGylation, O2 affinity and tissue oxygenation. AB - TetraPEGylated canine Hb, [SP (succinimidophenyl)-PEG5K]4-canine-Hb, with PEGylation at its four reactive cysteine residues (a111 and b93) has been prepared and characterized. The hydrodynamic volume and the molecular radius of (SP-PEG5K)4-canine-Hb are intermediate to those of di- and hexaPEGylated human Hb as expected. However, the COP (colloidal osmotic pressure) of tetraPEGylated canine Hb is closer to that of hexaPEGylated human Hb than to that of diPEGylated human Hb. The O2 affinity of tetraPEGylated canine Hb is higher than that of canine Hb and comparable with that of hexaPEGylated Hb. The O2 affinity of tetraPEGylated canine Hb is not responsive to the presence of DPG (diphosphoglycerate) or chloride, but it retains almost full response to L-35, an allosteric effector that interacts at the aa-end of the central cavity. The tetraPEGylated canine Hb is vasoinactive in hamster in 10% top load infusion studies. It is also essentially non-hypertensive in an extreme exchange haemodilution protocol in hamster just as di- and hexaPEGylated human Hb. The O2 delivery by tetraPEGylated canine Hb is comparable with that of hexaPEGylated Hb but not as efficient as diPEGylated Hb. These results demonstrate that PEGylation induced solution properties of PEG [poly(ethylene glycol)]-Hb conjugates are dictated by the level and chemistry of PEGylation and the interplay of these plays a critical role in tissue oxygenation. The studies imply the need to establish the right level (and/or pattern) of PEGylation and O2 affinity of Hb PEG adducts in designing O2-carrying plasma volume expanders, and this remains the primary challenge in the design of PEGylated Hb as blood substitutes. PMID- 17425517 TI - Promoting health literacy through the health education assessment project. AB - BACKGROUND: The Council of Chief State School Officers' State Collaborative on Assessment and Student Standards Health Education Assessment Project (SCASS-HEAP) allows states to pool financial and human resources to develop effective ready-to use health education assessment resources through a collaborative process. The purpose of this article is to describe the extensive ongoing development of the SCASS-HEAP and its benefits for important stakeholders in health and education. METHODS: A review of the products from the first decade of the SCASS-HEAP was undertaken. RESULTS: The SCASS-HEAP supports a comprehensive systems approach to helping educators focus effectively on the most important skills and issues in child and adolescent health and gives health education a place at the school reform table, providing visibility and credibility and promoting the essential links between health and learning. CONCLUSION: State education agencies and school districts can use SCASS-HEAP materials for assessment and, perhaps more importantly, to help teachers modify and improve instruction at the classroom level for increased student learning. PMID- 17425518 TI - Differences in the correlates of physical activity between urban and rural Canadian youth. AB - BACKGROUND: Despite the benefits of physical activity (PA), a significant proportion of youth remains inactive. Studies assessing differences in the correlates of PA among urban and rural youth are scarce, and such investigations can help identify subgroups of the population that may need to be targeted for special intervention programs. The purpose of this study was to assess differences in the correlates of PA between Canadian urban and rural youth. METHODS: The sample consisted of 1398 adolescents from 4 urban schools and 1290 adolescents from 4 rural schools. Mean age of the participants was 15.6 +/- 1.3 years. Hierarchical regression analyses were used to examine the association between self-reported PA and a number of demographic, psychological, behavioral, and social correlates. RESULTS: Common correlates between the 2 locations included gender (with girls being less active than boys) perceptions of athletic/physical ability, self-efficacy, interest in organized group activities, use of recreation time, and friends' and siblings' frequency of participation in PA. Active commuting to school and taking a physical education class were unique correlates of PA at the multivariate level in urban and rural students, respectively. Variance explained in PA ranged from 43% for urban school students to 38% for rural school students. CONCLUSIONS: Although more similarities than discrepancies were found in the correlates of PA between the 2 geographical locations, findings from this study strengthen the policies that argue for a coordinated multisector approach to the promotion of PA in youth, which include the family, school, and community. PMID- 17425519 TI - The use of portfolios in coordinated school health programs: benefits and challenges to implementation. AB - BACKGROUND: Coordinated school health programs (CSHP) frequently struggle with how to adequately evaluate implementation. The CSHP framework provides flexibility in how it is implemented; however, this flexibility makes it a challenge to effectively evaluate. Portfolios have been used as a technique for evaluating progress and achievement. This article describes the development and implementation of a CSHP portfolio and examines the perceptions of school personnel who have used CSHP portfolios. METHODS: This study employed content analysis and key informant interviews. Two elementary, 4 middle, and 2 high schools in Florida participated. RESULTS: Portfolios (N = 5) contained rich descriptions and program artifacts documenting each school's CSHP goals and activities. Key informant interviews (N = 14) revealed that school personnel found value in completing CSHP portfolios. CSHP portfolios were described as helpful in assessing progress, facilitating transition with new members, and building support for their efforts. Barriers to portfolio development included lack of time, money, and human resources. CONCLUSIONS: This study found the use of portfolios in documenting the implementation of the CSHP feasible and useful for school personnel. Portfolios provide a rich description of CSHP activities that may not be apparent through traditional program reports used for evaluation. As portfolios continue to be used in implementing CSHP, they may be viewed as a best process for CSHP implementation and a key element in the evaluation of CSHP. PMID- 17425520 TI - Sociodemographic differences in depressed mood: results from a nationally representative sample of high school adolescents. AB - BACKGROUND: Research on adolescent mental health suggests that prevalence rates for depressed mood are not uniformly distributed across all populations. This study examined demographic difference in depressed mood among a nationally representative sample of high school adolescents. METHODS: The 2003 National Youth Risk Behavior Survey was utilized to examine the association between depressed mood and demographic variables. To examine demographic associations, chi-square and follow-up logistic regression models were created for the sample of high school adolescents (N = 15,214). RESULTS: Caucasians and African Americans were significantly less likely to report depressed mood when compared to Hispanics and Others. Women were also more likely than men to report depressed mood. No significant differences were observed among levels of urbanicity and age. CONCLUSIONS: An understanding of the difference in the prevalence and correlates of depressed mood among adolescents of various ethnic/racial and gender groups deserves increased attention. Given the fact that challenges with depressed mood in adolescents can be substantial and relatively unrecognized, there is an increased need to identify these adolescents early and intervene with culturally appropriate interventions. PMID- 17425521 TI - Social influences, and attitudes and beliefs associated with smoking among border Latino youth. AB - BACKGROUND: Adolescent smoking rates have declined among all ethnic groups since the late 1990s. However, despite the recent declines and intervention efforts, today smoking remains a serious problem among youth, with a quarter of adolescents being current smokers by the time they complete 12th grade. This problem is particularly prevalent among Latino youth, who have among the highest rates of lifetime and past-30-day use. The purpose of this study was to examine the association between psychosocial factors and the smoking behavior of Latino youth living along the Mexico-US border. METHODS: A convenient sample of 2471 middle and high school Latino students was surveyed in fall 2000. Logistic regression analysis was used to examine the association between study risk factors and youth smoking behavior. RESULTS: The strongest predictor of lifetime and past-30-day smoking was peer influence; however, the strength of the association was greater with recent use. There were also differences in the influence of family and attitudes and beliefs between the 2 groups. CONCLUSIONS: These differences need to be taken into consideration to guide development of tailored prevention and control interventions aimed at this specific group. These efforts should address social influences to smoke, particularly those from peers; promote changes in attitudes and beliefs toward smoking; increase understanding of the addictive nature of nicotine; and provide development of skills young people need to resist social and environmental pressures to smoke. Strict control and enforcement measures are needed to completely eliminate the sale of cigarettes to minors. PMID- 17425522 TI - Use of brief interventions for drug abusing teenagers within a middle and high school setting. AB - BACKGROUND: Promising and encouraging results have been recently reported on the use of briefer interventions for adolescent drug abusers. Because middle- and high-school-based drug abuse intervention programs have grown in popularity over the past several decades, the use of brief interventions (BIs) in school settings merits consideration. METHODS: We review several clinical and school contextual issues pertaining to the scientific efficacy, feasibility, and application of BIs for students who are abusing drugs. RESULTS: Several advantages for employing BIs in a school setting are identified, including the relatively high base rate of students with mild-to-moderate drug involvement and the likelihood that school counselors can readily learn BI techniques. Caveats of implementing BIs include practical, systemic, and clinical barriers. CONCLUSIONS: Despite concerns, schools are a viable setting in which to screen youth for drug abuse problems and to conduct a BI. PMID- 17425523 TI - Association of the presence of state and district health education policies with school tobacco prevention program practices. AB - BACKGROUND: Texas does not require health education or tobacco use prevention education (TUPE) in its middle schools. During planning for the Texas Tobacco Prevention Initiative, this baseline research was conducted to (1) describe tobacco prevention and control practices in middle schools prior to the pilot, (2) analyze implementation of a state law prohibiting tobacco use on campuses and at school events, and (3) identify how schools are influenced by district policies requiring health education. METHODS: Written surveys derived from the 2000 School Health Education Profile Tobacco Module developed by the Centers for Disease Control and Prevention were completed by principals and health coordinators at schools in districts requiring health education (n = 31) and schools without district requirements (n = 32). RESULTS: School tobacco policy familiarity and enforcement were consistently reported in response to a state law with rigorous recommendations prohibiting tobacco use. Significantly more activity in numerous components of TUPE was reported in schools in districts with a health education requirement. CONCLUSIONS: Results have implications for intervention programs planned in schools located in states seeking to develop or strengthen state laws, or in states without health education or specific health content requirements. PMID- 17425525 TI - Fitts's law holds for action perception. AB - Fitts's law is one of the most well-established principles in psychology. It captures the relation between speed and accuracy in performed and imagined movements. The aim of this study was to determine whether this law also holds during the perception of other people's actions. Subjects were shown apparent motion displays of a person moving his arm between two identical targets. Target width, the separation between targets, and movement speed were varied. Subjects reported whether the person could move at the perceived speed without missing the targets. The movement times reported as being just possible were exactly those predicted by Fitts's law (r(2)= .96). A subsequent experiment demonstrated the same lawful relation for the perception of a robot arm (r(2)= .93). To our knowledge, this makes Fitts's law the first motor principle that holds in imagery and the perception of biological and non-biological agents. PMID- 17425526 TI - Dorsal and ventral processing under dual-task conditions. AB - It is widely acknowledged that visual input is processed along two anatomically and functionally distinct pathways--a ventral pathway for conscious perception and a dorsal pathway for action control. The present study investigated whether the apparent direct and unmediated processing in the dorsal stream is subject to capacity limitations. Specifically, we tested whether a simple dorsal task of grasping an object is affected by the psychological refractory period (PRP), a well-known indication of capacity limitations. Subjects performed an auditory choice reaction task and then, following varying delays, had to judge an object's width (ventral task) or grasp an object across its width (dorsal task). Although these tasks were differentially affected by irrelevant variation of the objects' length, they were subject to comparable dual-task interference. These results show that despite important differences between ventral and dorsal information processing, both modes of processing are constrained by limited capacities. PMID- 17425527 TI - Constructivist coding: learning from selective feedback. AB - Although much learning in real-life environments relies on highly selective feedback about outcomes, virtually all cognitive models of learning, judgment, and categorization assume complete and representative feedback. We investigated empirically the effect of selective feedback on decision making and how people code experience with selective feedback. The results showed that, in contrast to a commonly raised concern, performance was not impaired following learning with selective and biased feedback. Furthermore, even in a simple decision task, the experience that people acquired was not a mere recording of the observed outcomes, but rather a reconstruction from general task knowledge. PMID- 17425528 TI - Remembering can cause forgetting--but not in negative moods. AB - Repeated retrieval of a subset of previously observed events can cause forgetting of the non-retrieved events. We examined how affective states experienced during retrieval modulate such retrieval-induced forgetting by inducing positive, negative, and neutral moods in subjects immediately before they attempted to retrieve studied items. On the basis of recent work, we hypothesized that positive moods encourage relational processing, which should increase interference from related events and thus enhance retrieval-induced forgetting. By contrast, negative moods should encourage item-specific processing, which should reduce interference and thus reduce such forgetting. Our results are consistent with these predictions. When subjects were in negative moods, repeated retrieval did not cause forgetting of the non-retrieved material, whereas when subjects were in positive and neutral moods, they showed reliable retrieval induced forgetting. Our findings suggest that the emotions involved during interrogation of a witness can affect the result of repeated interrogations. PMID- 17425529 TI - Animals and androids: implicit associations between social categories and nonhumans. AB - People commonly ascribe lesser humanness to others than to themselves. Two senses of humanness appear to be involved: attributes that are unique to humans and those that constitute essential "human nature." Denying uniquely human and human nature attributes to other people may implicitly liken them to animals and automata, respectively. In the present study, the go/no-go association task was used to assess implicit associations among social categories exemplifying the two senses of humanness, traits representing these senses, and the two types of nonhumans. Congruent associations (among artists, human-nature traits, and animals; among businesspeople, uniquely human traits, and automata) were consistently stronger than incongruent associations. Explicit ratings supported these differential associations. Social perception may involve two subtle ways of dehumanizing others. PMID- 17425530 TI - Better the DVL you know: acronyms reveal the contribution of familiarity to single-word reading. AB - Current theories of reading are divided between dual-route accounts, which propose that separable processes subserve word recognition for orthographically regular and irregular strings, and connectionist models, which propose a single mechanism mapping form to meaning. These theories make distinct predictions about the processing of acronyms, which can be orthographically illegal and yet familiar, as compared with the processing of pseudowords, which are regular but unfamiliar. This study examined whether acronyms are processed like pseudowords and words. Event-related potentials (ERPs) were recorded as subjects viewed familiar and unfamiliar acronyms, words, pseudowords, illegal strings, and--as the targets of the substantive behavioral task--proper names. Familiar acronyms elicited repetition effects on the N400 component, a functionally specific index of semantic activation processes; repetition effects for familiar acronyms were similar in magnitude, timing, and scalp distribution to those for words and pseudowords. The similarity of the brain response to familiar--but--illegal and unfamiliar--but--legal classes of stimuli is inconsistent with predictions of dual-route models of reading. PMID- 17425531 TI - Changes in anterior cingulate and amygdala after cognitive behavior therapy of posttraumatic stress disorder. PMID- 17425532 TI - Is automatic emotion regulation associated with agreeableness? A perspective using a social neuroscience approach. PMID- 17425533 TI - The Wickelgren power law and the Ebbinghaus savings function. PMID- 17425534 TI - Syntactic priming in comprehension: evidence from event-related potentials. AB - Syntactic priming is the facilitation of processing that occurs when a sentence has the same syntactic form as a preceding sentence. Such priming effects have been less consistently demonstrated in comprehension than in production, and those that have been reported have depended on the repetition of verbs across sentences. In an event-related potential experiment, subjects read target sentences containing reduced-relative clauses. Each was preceded by a sentence that contained the same verb and either a reduced-relative or a main-clause construction. Reduced-relative primes elicited a larger positivity than did main clause primes. Reduced-relative targets that were preceded by a main-clause prime elicited a greater positivity than the same target sentences following a reduced relative prime. In addition, syntactic priming effects were dissociated from effects of lexical repetition at the verb. PMID- 17425535 TI - Distinguishing the neural correlates of episodic memory encoding and semantic memory retrieval. AB - Episodic memory and semantic memory interact very closely. In particular, episodic memory encoding (EE) tends to elicit semantic memory retrieval (SR), and vice versa. Thus, similar activations for EE and SR in functional neuroimaging studies may reflect shared memory processes, or they may reflect the fact that EE and SR are usually confounded. To address this issue, we used a factorial functional magnetic resonance imaging approach to disentangle the neural correlates of EE and SR. Within the left temporal lobe, the hippocampus was associated with successful EE, whereas a posterior lateral region was associated with successful SR. Within the left inferior prefrontal cortex, a posterior region was involved in SR, a mid region was involved in both SR and EE, and an anterior region was involved in EE, but only when SR was also high. Thus, the neural correlates of EE and SR are dissociable but interact in specific brain regions. PMID- 17425536 TI - Rubber hands feel the touch of light. AB - Two experiments involving a total of 220 subjects are reported. The experiments document that "stroking" a false hand with the bright beam of light from a laser pointer can produce tactile and thermal sensations when the hand can be seen as one's own. Overall, 66% of subjects reported somatic sensations from the light. Felt hand location was recalibrated toward the location of the false hand for those subjects who felt the light. Moreover, the proprioceptive recalibration from the laser experience was comparable to that produced by actual coordinated brushing of the false hand and of the unseen real hand after 2 min of stimulation. The illusion may be experienced on one's real hand as well. The results are discussed in terms of multisensory integration. PMID- 17425537 TI - Haptic recognition of static and dynamic expressions of emotion in the live face. AB - If humans can detect the wealth of tactile and haptic information potentially available in live facial expressions of emotion (FEEs), they should be capable of haptically recognizing the six universal expressions of emotion (anger, disgust, fear, happiness, sadness, and surprise) at levels well above chance. We tested this hypothesis in the experiments reported here. With minimal training, subjects' overall mean accuracy was 51% for static FEEs (Experiment 1) and 74% for dynamic FEEs (Experiment 2). All FEEs except static fear were successfully recognized above the chance level of 16.7%. Complementing these findings, overall confidence and information transmission were higher for dynamic than for corresponding static faces. Our performance measures (accuracy and confidence ratings, plus response latency in Experiment 2 only) confirmed that happiness, sadness, and surprise were all highly recognizable, and anger, disgust, and fear less so. PMID- 17425538 TI - Mind-set matters: exercise and the placebo effect. AB - In a study testing whether the relationship between exercise and health is moderated by one's mind-set, 84 female room attendants working in seven different hotels were measured on physiological health variables affected by exercise. Those in the informed condition were told that the work they do (cleaning hotel rooms) is good exercise and satisfies the Surgeon General's recommendations for an active lifestyle. Examples of how their work was exercise were provided. Subjects in the control group were not given this information. Although actual behavior did not change, 4 weeks after the intervention, the informed group perceived themselves to be getting significantly more exercise than before. As a result, compared with the control group, they showed a decrease in weight, blood pressure, body fat, waist-to-hip ratio, and body mass index. These results support the hypothesis that exercise affects health in part or in whole via the placebo effect. PMID- 17425539 TI - Postidentification feedback affects real eyewitnesses. AB - Many studies of simulated eyewitness situations have shown that under certain laboratory conditions, people's confidence about their identifications predicts their accuracy, but that their reported confidence can be affected by telling them that they chose the suspect. In this study, eyewitnesses (n= 134) to real crimes took part in lineups at an identification suite in the United Kingdom and were asked questions about their memory both before and after they were told whether they had identified the suspect or a filler. Before the eyewitnesses were told whether they had identified the suspect or a filler, their responses to several questions reliably differentiated between those who identified the suspect and those who identified a filler. In addition, responses to the memory questions were affected by telling the eyewitnesses whether or not they had identified the suspect. These results show that postidentification feedback affects real eyewitnesses and highlight the importance of recording meta-memory variables before an eyewitness discovers whether he or she has identified the suspect. PMID- 17425540 TI - When looks are everything: appearance similarity versus kind information in early induction. AB - The goal of this research was to examine mechanisms underlying early induction- specifically, the relation between induction and categorization. Some researchers argue that even early in development, induction is based on category-membership information, whereas others argue that early induction is based primarily on similarity. Children 4 and 5 years of age participated in two types of tasks: categorization and induction. Both tasks were performed with artificial animal like categories in which appearance was pitted against category membership. Although the children readily acquired category-membership information and subsequently used this information in categorization tasks, they ignored category membership during the induction task, relying instead on the appearance of items. These results support the idea that early in development, induction is similarity based. PMID- 17425541 TI - Children's understanding and experience of mixed emotions. AB - Though some models of emotion contend that happiness and sadness are mutually exclusive in experience, recent findings suggest that adults can feel happy and sad at the same time in emotionally complex situations. Other research has shown that children develop a better conceptual understanding of mixed emotions as they grow older, but no research has examined children's actual experience of mixed emotions. To examine developmental differences in the experience of mixed emotions, we showed children ages 5 to 12 scenes from an animated film that culminated with a father and daughter's bittersweet farewell. In subsequent interviews, older children were more likely than younger children to report experiencing mixed emotions. These results suggest that in addition to having a better conceptual understanding of mixed emotions, older children are more likely than younger children to actually experience mixed emotions in emotionally complex situations. PMID- 17425543 TI - OASIS wound matrix versus Hyaloskin in the treatment of difficult-to-heal wounds of mixed arterial/venous aetiology. AB - Mixed arterial/venous (A/V) ulcers are difficult to treat and slow to heal likely as a result of deficiencies in molecular and cellular elements in the wound bed. Recently, biomaterials have been developed that replace extracellular matrix (ECM) molecules and growth factors critical to the normal healing process. In this study, the effects of OASIS and Hyaloskin were evaluated to compare the effectiveness of these two ECM-based products in their ability to achieve complete wound healing of mixed A/V ulcers. After 16 weeks of treatment, patients in each group were evaluated on four criteria: complete wound healing, time to dressing change, pain and comfort. Complete wound closure was achieved in 82.6% of OASIS-treated ulcers compared with 46.2% of Hyaloskin-treated ulcers (P < 0.001). Statistically significant differences favouring the OASIS treatment group were also reported for time to dressing change (P < 0.05), pain (P < 0.05) and patient comfort (P < 0.01). Overall, OASIS was superior to Hyaloskin for the treatment of patients with mixed A/V ulcers, a population in which standard treatment options largely consist of moist wound dressings and compression therapy is typically not an option. OASIS is a useful and well-tolerated treatment for mixed A/V ulcers that has the potential to improve quality of life and reduce costs associated with standard of care. PMID- 17425544 TI - Successful management of deep facial burns in a patient with extensive third degree burns: the role of a nanocrystalline silver dressing in facilitating resurfacing. AB - Full-thickness burns of the face are notoriously difficult to repair and their management poses a series of problems to the surgeon. We present the case of a 49 year-old man (only survivor of a catastrophic airport accident) with third-degree flame burns to >80% of total body surface area and extensive face damage who achieved a fully satisfactory outcome after a treatment plan based on gradual escharectomy followed by application of artificial dermis and, later, grafting with sheets of cultured keratinocytes. Re-epithelialisation was already visible at day 16 after admission and all facial wounds were closed by day 56, the treatment continuing on the scalp. Within 6 months of the accident, the patient had recovered functional and cosmetic features (including re-growth of skin appendages) that were beyond expectations. The use of nanocrystalline silver coated dressings during the escharectomy and resurfacing phases was important, as part of a multifaceted strategy, in ensuring excellent antimicrobial control, thus avoiding the need for autologous grafting and contributing to a rapid healing and complete restoration of the face and head skin. PMID- 17425545 TI - Paediatric partial-thickness scald burns--is Biobrane the best treatment available? AB - The article critically analyses the role of Biobrane, one of the widely used modern biosynthetic dressings, in paediatric partial-thickness scald burns. Most of the studies, available in the literature, confirmed that Biobrane has significantly reduced the hospital stay, wound healing time and requirements of pain medications. However, very few studies with long-term follow up are available. Further randomised controlled trials are required to challenge the supremacy of Biobrane in paediatric partial-thickness burns. PMID- 17425546 TI - Wound bed preparation of difficult wounds: an evolution of the principles of TIME. AB - In the last few years, considerable progress has been made in the treatment of chronic ulcers, thanks to new therapy methods. Wound bed preparation is a modern approach for the removal of local barriers to healing by optimising debridement, reduction of bioburden and exudate management through the TIME principles, which have been introduced by the International Advisory Board on Wound Bed Preparation. However, this protocol does not evaluate the state of the repair process and therefore does not suggest the ideal therapeutic choice for each single patient. The revised TIME-H concept considers also the supposed healing time, H, and gives a score that correlates the wound condition with the incidental concomitance with medical pathologies related to the therapeutic measures, thus guiding the clinician towards a practical and systematic approach in the treatment. By applying this scheme to our situation, the average healing time was considerably reduced. The formulation of the new protocol TIME-H for a critical assessment of treatment scheme, which also includes the general conditions of the patient, represents a more rational and adequate approach for an accurate prognosis and therefore for a more suitable therapeutic choice in the treatment of difficult wounds. PMID- 17425547 TI - Clinical predictors of treatment failure for diabetic foot infections: data from a prospective trial. AB - To aid clinicians in selecting the appropriate approach for treating patients with diabetic foot infections, we investigated whether any baseline clinical findings predicted an unfavourable clinical outcome. Using data from a large, prospective treatment trial of diabetic foot infections (SIDESTEP), we assessed the association between clinical treatment failure and baseline history, physical and laboratory findings, by univariate and multivariate logistic regression analyses. Among 402 patients clinically evaluable 10 days after completing antibiotic therapy, baseline factors significantly (P < 0.05) associated by univariate analysis with treatment failure were 'severe' (versus 'moderate') University of Texas (UT) wound grade; elevated white blood cell count, C-reactive protein or erythrocyte sedimentation rate; high wound severity score; inpatient treatment; low serum albumin; male sex; and skin temperature of affected foot >10 degrees C above that of unaffected foot. By multivariate logistic regression only severe UT wound grade (odds ratio 2.1) and elevated white blood cell count [odds ratio 1.7 for a 1 standard deviation (2971 cells/mm(3)) increase] remained statistically significant. Clinical failure rates were 46% for patients with both risk factors compared with 10% for patients with no risk factors and 16-17% for patients with one risk factor. Increased white blood cell count and severe UT wound grade at baseline, but not other features, were significant independent and additive risk factors for clinical failure in patients treated for a diabetic foot infection. PMID- 17425548 TI - The role of vascular endothelial growth inhibitor in wound healing. AB - Vascular endothelial growth inhibitor (VEGI) is an endothelial cell-specific cytokine and a potent inhibitor of endothelial cell proliferation and angiogenesis. The role of VEGI in angiogenesis related to tissue repair has previously not been investigated. Biopsies from different wound types were analysed by immunohistochemistry and quantitative polymerase chain reaction for the presence of VEGI protein and transcript, respectively. Human vascular endothelial cell line was transfected with VEGI expression plasmid and tested for their in vitro angiogenesis properties. Immunohistochemical staining for VEGI showed reduced expression in the dermal layer of the acute wounds compared with the chronic wound or normal skin. The ability of VEGI to prevent angiogenesis by in vitro assays showed that VEGI acts as a suppressor to the proliferation and microtubule formation of endothelial cells, and the addition of Hepatocyte Growth Factor had little effect on the ability of cell lines expressing the VEGI gene to increase microtubule formation. The aberrant expression of VEGI in different wound types appears to be linked to the outcome of the healing in these wounds. The altered expression of VEGI in chronic wounds constitutes an important target of future therapies. PMID- 17425549 TI - Randomised clinical trial of Hydrofiber dressing with silver versus povidone iodine gauze in the management of open surgical and traumatic wounds. AB - This prospective, randomised clinical trial compared pain, comfort, exudate management, wound healing and safety with Hydrofiber dressing with ionic silver (Hydrofiber Ag dressing) and with povidone-iodine gauze for the treatment of open surgical and traumatic wounds. Patients were treated with Hydrofiber Ag dressing or povidone-iodine gauze for up to 2 weeks. Pain severity was measured with a 10 cm visual analogue scale (VAS). Other parameters were assessed clinically with various scales. Pain VAS scores decreased during dressing removal in both groups, and decreased while the dressing was in place in the Hydrofiber Ag dressing group (n = 35) but not in the povidone-iodine gauze group (n = 32). Pain VAS scores were similar between treatment groups. At final evaluation, Hydrofiber Ag dressing was significantly better than povidone-iodine gauze for overall ability to manage pain (P < 0.001), overall comfort (P < or = 0.001), wound trauma on dressing removal (P = 0.001), exudate handling (P < 0.001) and ease of use (P < or = 0.001). Rates of complete healing at study completion were 23% for Hydrofiber Ag dressing and 9% for povidone-iodine gauze (P = ns). No adverse events were reported with Hydrofiber Ag dressing; one subject discontinued povidone-iodine gauze due to adverse skin reaction. Hydrofiber Ag dressing supported wound healing and reduced overall pain compared with povidone-iodine gauze in the treatment of open surgical wounds requiring an antimicrobial dressing. PMID- 17425550 TI - Negative pressure wound therapy via vacuum-assisted closure following partial foot amputation: what is the role of wound chronicity? AB - Randomised clinical trials (RCTs) to evaluate diabetic foot wound therapies have systematically eliminated large acute wounds from evaluation, focusing only on smaller chronic wounds. The purpose of this study was to evaluate the proportion and rate of wound healing in acute and chronic wounds after partial foot amputation in individuals with diabetes treated with negative pressure wound therapy (NPWT) delivered by the vacuum-assisted closure (VAC) device or with standard wound therapy (SWT). This study constitutes a secondary analysis of patients enrolled in a 16-week RCT of NPWT: 162 open foot amputation wounds (mean wound size = 20.7 cm(2)) were included. Acute wounds were defined as the wounds less than 30 days after amputation, whereas chronic wounds as the wounds greater than 30 days. Inclusion criteria consisted of individuals older than 18 years, presence of a diabetic foot amputation wound up to the transmetatarsal level and adequate perfusion. Wound size and healing were confirmed by independent, blinded wound evaluators. Analyses were done on an intent-to-treat basis. There was a significantly higher proportion of acute wounds (SWT = 59; NPWT = 63) than chronic wounds (SWT = 26; NPWT = 14), evaluated in this clinical trial (P = 0.001). There was no significant difference in the proportion of acute and chronic wounds achieving complete wound closure in either treatment group. Despite this finding, the Kaplan-Meier curves demonstrated statistically significantly faster healing in the NPWT group in both acute (P = 0.030) and chronic wounds (P = 0.033). Among the patients treated with NPWT via the VAC, there was not a significant difference in healing as a function of chronicity. In both the acute and the chronic wound groups, results for patients treated with NPWT were superior to those for the patients treated with SWT. These results appear to indicate that wound duration should not deter the clinician from using this modality to treat complex wounds. PMID- 17425551 TI - Negative pressure wound therapy: treating a venomous insect bite. AB - Reports of spider bites appear throughout North America. Bites associated with the brown recluse spider (Loxosceles recluse) cause serious medical complications because the venom of this spider contains a powerful necrotising agent with the potential to cause severe cutaneous necrosis. Although not much is known regarding the application of negative pressure wound therapy (NPWT) to spider bites, this therapy has considerable literature support for its efficacy, cost effectiveness and ease of use in chronic, difficult-to-heal wounds. A case study using NPWT to successfully treat a non healing upper arm wound presumed due to a venomous spider bite is presented here. The patient was successfully treated with a new, less costly NPWT product called the Versatile 1 and a new combination drain plus dressing called the Miller DermiVex drain, both manufactured by Blue Sky Medical (Carlsbad, CA). PMID- 17425552 TI - Gross dimensional assessment of cutaneous wounds as an indicator of healing: importance of careful consideration. PMID- 17425553 TI - A different approach to the management of necrotizing fasciitis in neonates: hyperbaric oxygen. PMID- 17425554 TI - Cortical control of eye and head movements: integration of movements and percepts. AB - The cortical control of eye movements is well known. It remains unclear, however, as to how the eye fields of the frontal lobes generate and coordinate eye and head movements. Here, we review the recent advances in electrical stimulation studies and evaluate relevant models. As electrical stimulation is conducted in head-unrestrained, behaving subjects with the evoked eye and head movements sometimes being indistinguishable from natural gaze shifts, a pertinent question becomes whether these movements are evoked by motor programs or sensory percepts. Recent stimulation studies in the visual cortex and the eye fields of the frontal lobes have begun to bring both possibilities to light. In addition, cognitive variables often interact with behavioral states that can affect movements evoked by stimulation. Identifying and controlling these variables are critical to our understanding of experimental results based on electrically evoked movements. This understanding is needed before one can draw inferences from such results to elucidate the neural mechanisms underlying natural and complex movements. PMID- 17425555 TI - The fidelity of the cortical retinotopic map in human amblyopia. AB - To delineate the fidelity of the functional cortical organization in humans with amblyopia, we undertook an investigation into how spatial information is mapped across the visual cortex in amblyopic observers. We assessed whether the boundaries of the visual areas controlled by the amblyopic and fellow fixing eye are in the same position, the fidelity of the retinotopic map within different cortical areas and the average receptive field size in different visual areas. The functional organization of the visual cortex was reconstructed using a fMRI phase-encoded retinotopic mapping analysis. This method sequentially stimulates each point in the visual field along the axes of a polar-coordinate system, thereby reconstructing the representation of the visual field on the cortex. We found that the cortical areas were very similar in normals and amblyopes, with only small differences in boundary positions of different visual areas between fixing and fellow amblyopic eye activation. Within these corresponding visual areas, we did find anomalies in retinotopy in some but not all amblyopes that were not simply a consequence of the poorer functional responses and affected central and peripheral field regions. Only a small increase in the average (or collective) receptive field size was found for full-field representation in amblyopes and none at all for central field representation. The former may simply be a consequence of the poorer functional responses. PMID- 17425556 TI - Long-term potentiation of neuronal excitation by neuron-glia interactions in the rat spinal dorsal horn. AB - By imaging neuronal excitation in rat spinal cord slices with a voltage-sensitive dye, we examined the role of glial cells in the P2X receptor agonist alphabeta methylene ATP (alphabetameATP)-triggered long-term potentiation (LTP) in the dorsal horn. Bath application of alphabetameATP potentiated neuronal excitation in the superficial dorsal horn. The potentiation was inhibited in the presence of the P2X receptor antagonists TNP-ATP, PPADS and A-317491, and was not induced in slices taken from rats neonatally treated with capsaicin. These results suggest that alphabetameATP acts on P2X receptors, possibly P2X(3) and/or P2X(2/3), in capsaicin-sensitive primary afferent terminals. Furthermore, the potentiation was inhibited by treatment with the glial metabolism inhibitor monofluoroacetic acid. Results obtained with the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB203580, tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL) 6, and antibodies to TNF-alpha and IL-6, as well as by double immunolabelling of activated p38 MAPK with markers of astrocytes and microglia, demonstrated that alphabetameATP activated p38 MAPK in astrocytes, and that the presence of proinflammatory cytokines and p38 MAPK activation were necessary for the induction of alphabetameATP-triggered LTP. These findings indicate that glial cells contribute to the alphabetameATP-induced LTP, which might be part of a cellular mechanism for the induction of persistent pain. PMID- 17425557 TI - Presynaptic GABA-A receptors prevent depression of nicotinic transmission in rabbit coeliac ganglion neurones. AB - We investigated the involvement of GABA-A receptors in the modulation of the nicotinic transmission of central origin in isolated rabbit coeliac ganglia. Our study was performed in vitro and the electrical activity of the ganglionic neurones was recorded using intracellular recording techniques. During iterative stimulation of the splanchnic nerves, the synaptic action potential probability decreased gradually, indicating a depression of the nicotinic activation. Pharmacological agents acting at GABA-A receptors modulated the action potential probability during the train of pulses. Muscimol (a GABA-A receptor agonist), diazepam (a benzodiazepine site agonist) and 1-[2 [[(diphenylmethylene)imino]oxy]ethyl]-1,2,5,6-tetrahydro-3-pyridinecarboxylic acid hydrochloride (a GABA uptake blocker) increased this probability. Conversely, gabazine or bicuculline (two GABA-A receptor antagonists), picrotoxin (a picrotoxin site agonist) and flumazenil (a benzodiazepine site antagonist) reduced it. These results demonstrate that endogenous GABA, released during the train of pulses, facilitates the central nicotinic activation of the ganglionic neurones by acting on GABA-A receptors. Muscimol also reduced the amplitude ratio of excitatory postsynaptic potentials triggered during the paired-pulse protocol without any change in postsynaptic properties. This result is consistent with a presynaptic action of GABA-A receptors. Our study shows that presynaptic GABA-A receptors facilitate the central nicotinic activation of prevertebral ganglionic neurones and thus play a novel role in the integrative properties of the sympathetic prevertebral ganglia. PMID- 17425558 TI - Functional and ultrastructural analysis of group I mGluR in striatal fast-spiking interneurons. AB - Striatal parvalbumin-containing fast-spiking (FS) interneurons provide a powerful feedforward GABAergic inhibition on spiny projection neurons, through a widespread arborization and electrical coupling. Modulation of FS interneuron activity might therefore strongly affect striatal output. Metabotropic glutamate receptors (mGluRs) exert a modulatory action at various levels in the striatum. We performed electrophysiological recordings from a rat striatal slice preparation to investigate the effects of group I mGluR activation on both the intrinsic and synaptic properties of FS interneurons. Bath-application of the group I mGluR agonist, (S)-3,5-dihydroxyphenylglycine (3,5-DHPG), caused a dose dependent depolarizing response. Both (S)-(+)-alpha-amino-4-carboxy-2 methylbenzeneacetic acid (LY367385) and 7-(hydroxyimino)cyclopropa[b]chromen-1a carboxylate ethyl ester (CPCCOEt), selective mGluR1 antagonists, significantly reduced the amplitude of the membrane depolarization caused by 3,5-DHPG application. Conversely, mGluR5 antagonists, 2-methyl-6-(phenylethylnyl)pyridine hydrochloride (MPEP) and 6-methyl-2-(phenylazo)-3-pyridinol (SIB1757), were unable to affect the response to 3,5-DHPG, suggesting that only mGluR1 contributes to the 3,5-DHPG-mediated excitatory action on FS interneurons. Furthermore, mGluR1 blockade significantly decreased the amplitude of the glutamatergic postsynaptic potentials, whereas the mGluR5 antagonist application produced a small nonsignificant inhibitory effect. Surprisingly, our electron microscopic data demonstrate that the immunoreactivity for both mGluR1a and mGluR5 is expressed extrasynaptically on the plasma membrane of parvalbumin immunoreactive dendrites of FS interneurons. Together, these results suggest that despite a common pattern of distribution, mGluR1 and mGluR5 exert distinct functions in the modulation of FS interneuron activity. PMID- 17425559 TI - Protein tyrosine phosphatase inhibition reduces degeneration of dopaminergic substantia nigra neurons and projections in 6-OHDA treated adult rats. AB - The survival of injured adult dopaminergic substantia nigra pars compacta neurons can be promoted by various neurotrophic factors. Most neurotrophic factor receptors are activated by intracellular tyrosine phosphorylation upon ligand binding and are subsequently inactivated or dephosphorylated by protein tyrosine phosphatases. This raised the possibility that tyrosine phosphatase inhibition might improve neuronal survival. Here, we infused the stable water-soluble tyrosine phosphatase-specific inhibitor, peroxovanadium [potassium bisperoxo(1,10 phenanthroline)oxovanadate (V) (bpV(phen))], for 14 days close to the substantia nigra starting immediately after a unilateral moderate injury by injection of the neurotoxin 6-hydroxydopamine (6-OHDA) into the midbrain of adult Sprague-Dawley rats. The dopaminergic nigrostriatal neurons were identified by retrograde tracing with fluorogold 7 days prior to the injury. With infusion of 3 or 10 microm peroxovanadium, 75% of these neurons survived compared to 45% in vehicle infused rats. Degeneration of the dopaminergic projections to the neostriatum was also reduced by 10 microm peroxovanadium. Twenty minutes after an acute injection of peroxovanadium into the substantia nigra, increased tyrosine phosphorylation in Western blots of nigral extracts was seen in the same protein bands as after injections of brain-derived neurotrophic factor (BDNF) or NT-4. This suggests that peroxovanadium enhances endogenous neurotrophic signalling resulting in improved neuronal survival. The neuroprotective effects of this small molecule protein tyrosine phosphatase inhibitor represent a proof-of-principle for a novel treatment strategy in a model for Parkinson's disease. PMID- 17425560 TI - Phenylalanine activates the mitochondria-mediated apoptosis through the RhoA/Rho associated kinase pathway in cortical neurons. AB - Phenylketonuria (PKU) is caused by deficiency of phenylalanine hydroxylase, resulting in an accumulation of phenylalanine in brain tissue and cerebrospinal fluid of phenylketonuria patients. Phenylketonuria is neuropathologically characterized by neuronal cell loss, white matter abnormalities, dendritic simplification, and synaptic density reduction. The neuropathological effect may be due to the "toxicity" of the high concentration of phenylalanine, while the underlying mechanism remains unclear. In this study, we found that cultured cerebral cortical neurons underwent mitochondria-mediated apoptosis when exposed to phenylalanine. We further demonstrated that phenylalanine induced RhoA activation. Phenylalanine also promoted myosin light chain (MLC) phosphorylation, which might be the result of the activation of Rho-associated kinase (ROCK). The RhoA antagonist, C3 transferase (C3), Rho-associated kinase specific inhibitor, Y 27632, and the overexpression of either dominant negative RhoA or dominant negative Rho-associated kinase inhibited phenylalanine-induced caspase-3 activation and rescued neurons from apoptosis, indicating that the RhoA/Rho associated kinase signalling pathway plays an important role in phenylalanine induced neuronal apoptosis. PMID- 17425561 TI - Radiation-induced apoptosis in retinal progenitor cells is p53-dependent with caspase-independent DNA fragmentation. AB - Caspases are important executioners of the endogenous cell death program. However, their function is not restricted to the induction of cell death. Caspases may process cytokines and contribute to cell differentiation or lymphocyte proliferation. In addition to their pleiotropic functions we show evidence that, under certain conditions, caspases are activated during apoptosis without executing the cell death program. Following whole body irradiation, p53 and caspases were activated in both the cerebellum and eye of postnatal day 5 mice. Although p53 activation and cell death kinetics were similar in both the cerebellum and eye, the processing of caspases was protracted and reduced in the eye. In particular, retinal caspase activation appeared not to be the executioner of cell death; incubation of retinal and cerebellar explants in the presence of the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone prevented DNA fragmentation, a hallmark of apoptosis, only in cerebellar granule cells. In contrast, in retinal cells no impairment of DNA fragmentation was observed in the presence of N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone, indicating p53-dependent but caspase-independent cell death pathways despite caspase activation. PMID- 17425562 TI - The extreme C-terminus of GluRdelta2 is essential for induction of long-term depression in cerebellar slices. AB - Long-term depression (LTD) of parallel fibre (PF)-Purkinje cell synapses in the cerebellum is recognized as a cellular substrate of motor learning. Although the delta2 glutamate receptor (GluRdelta2) has been shown to be crucial for LTD, the mechanisms by which GluRdelta2 functions remain elusive. In this study, we developed a virus vector-based gene transfer approach to rescue impaired LTD in GluRdelta2-null Purkinje cells in cerebellar slice preparations. We demonstrated that LTD was restored in GluRdelta2-null Purkinje cells transduced with wild-type but not with mutant GluRdelta2, which lacked the PDZ-ligand domain in the C terminus. Immunohistochemical analysis revealed no difference in expression levels or spine localization patterns between virally introduced wild-type and mutant GluRdelta2 proteins. Similarly, LTD was abrogated in Purkinje cells that had been acutely perfused with peptides, hampering the interaction of GluRdelta2 with PDZ proteins such as PSD-93, PTPMEG and S-SCAM but not with delphilin. Together, these results indicate that PDZ proteins that bind to the C-terminus of GluRdelta2 are not essential for localizing GluRdelta2 at synapses but are crucial for conveying signals necessary for the induction of LTD. PMID- 17425563 TI - Proliferative responses to growth factors decline rapidly during postnatal maturation of mammalian hair cell epithelia. AB - Millions of lives are affected by hearing and balance deficits that arise as a consequence of sensory hair cell loss. Those deficits affect mammals permanently, but hearing and balance recover in nonmammals after epithelial supporting cells divide and produce replacement hair cells. Hair cells are not effectively replaced in mammals, but balance epithelia cultured from the ears of rodents and adult humans can respond to hair cell loss with low levels of supporting cell proliferation. We have sought to stimulate vestibular proliferation; and we report here that treatment with glial growth factor 2 (rhGGF2) yields a 20-fold increase in cell proliferation within sheets of pure utricular hair cell epithelium explanted from adult rats into long-term culture. In epithelia from neonates, substantially greater proliferation responses are evoked by rhGGF2 alone, insulin alone and to a lesser degree by serum even during short-term cultures, but all these responses progressively decline during the first 2 weeks of postnatal maturation. Thus, sheets of utricular epithelium from newborn rats average > 40% labelling when cultured for 72 h with bromo-deoxyuridine (BrdU) and either rhGGF2 or insulin. Those from 5- and 6-day-olds average 8-15%, 12-day-olds average < 1% and after 72 h there is little or no labelling in epithelia from 27- and 35-day-olds. These cells are the mammalian counterparts of the progenitors that produce replacement hair cells in nonmammals, so the postnatal quiescence described here is likely to be responsible for at least part of the mammalian ear's unique vulnerability to permanent sensory deficits. PMID- 17425564 TI - Synapse-specific localization of vesicular glutamate transporters in the rat olfactory bulb. AB - Vesicular glutamate transporters (VGLUTs) mediate the packaging of the excitatory neurotransmitter glutamate into synaptic vesicles. Three VGLUT subtypes have so far been identified, with distinct expression patterns in the adult brain. Here, we investigated the spatial distribution of the three VGLUTs in the rat olfactory bulb, a brain region containing a variety of glutamate synapses, both axodendritic and dendrodendritic. Using multilabelling confocal microscopy and electron microscopic immunocytochemistry, we showed that each VGLUT isoform has a highly selective localization in olfactory bulb synapses. VGLUT1 is present at dendrodendritic synapses established by the output neurones (mitral and tufted cells) with bulbar interneurones in the glomerular layer and external plexiform layer, as well as in axonal synapses of the granule cell layer. By contrast, VGLUT2 is strongly expressed in axon terminals of olfactory sensory neurones, which establish synapses with second-order neurones in the glomerular neuropil. VGLUT2 is also found in the outer part of the external plexiform layer and in the granule cell layer but colocalizes only partially with VGLUT1. Finally, we showed that VGLUT3 is exclusively located in the glomerular neuropil, where it colocalizes extensively with the vesicular inhibitory amino acid transporter vesicular GABA transporter, suggesting that it is associated with a subset of inhibitory synapses. Together, these observations extend previous findings on VGLUT distribution in the forebrain, and suggest that each VGLUT subtype has a specific function in the distinct features of axodendritic and dendrodendritic synapses that characterize the olfactory bulb circuit. PMID- 17425565 TI - Axons of callosal neurons bifurcate transiently at the white matter before consolidating an interhemispheric projection. AB - The main alternative output routes of adult cortical axons are the internal capsule and the corpus callosum. How do callosal axons choose their trajectories? We hypothesized that bifurcation followed by elimination of one branch is a developmental strategy for accomplishing this aim. Using embryonic and postnatal mice, we labelled cortical projecting neurons and quantified their axonal bifurcations in correlation with the mediolateral position of their somata. Bifurcating axons were numerous in the younger brains but declined during further development. Most bifurcating axons pertained to neurons located in the dorsolateral cortex. Moreover, callosal neurons bifurcate more often than subcortically projecting cells. We then quantified bifurcations formed by dissociated green fluorescent cells plated onto cortical slices. Cells grown over dorsolateral cortex bifurcated more often than those grown over medial cortex, irrespective of their positional origin in the donor. Removal of intermediate targets from the slices prevented bifurcation. We concluded that transient bifurcation and elimination of the lateral branch is a strategy employed by developing callosal axons in search of their targets. As cell body position and intermediate targets determine axon behaviour, we suggest that bifurcations are regulated by cues expressed in the environment. PMID- 17425566 TI - Effect of binocular retinal lesions on CRMP2 and CRMP4 but not Dyn I and Syt I expression in adult cat area 17. AB - Removal of retinal input from a restricted region of adult cat visual cortex leads to a substantial reorganization of the retinotopy within the sensory deprived cortical lesion projection zone (LPZ). Still little is known about the molecular mechanisms underlying this cortical map reorganization. We chose two members of the collapsin response mediator protein (CRMP) family, CRMP2 and CRMP4, because of their involvement in neurite growth, and compared gene and protein expression levels between normal control and reorganizing visual cortex upon induction of central retinal lesions. Parallel analysis of Dynamin I (Dyn I) and Synaptotagmin I (Syt I), two molecules implicated in the exocytosis endocytosis cycle, was performed because changes in neurotransmitter release have been implicated in cortical plasticity. Western blotting and real-time polymerase chain reaction revealed a clear time-dependent effect of retinal lesioning on CRMP2 and CRMP4 expression, with maximal impact 2 weeks post-lesion. Altered CRMP levels were not a direct consequence of decreased visual activity in the LPZ as complete surgical removal of retinal input to one hemisphere had no effect on CRMP2 or CRMP4 expression. Thus, CRMP expression is correlated to cortical reorganization following partial deafferentation of adult visual cortex. In contrast, Dyn I and Syt I were not influenced and thereby do not promote exocytosis-endocytosis cycle modifications in adult cat cortical plasticity. PMID- 17425567 TI - Dissociation and trafficking of rat GABAB receptor heterodimer upon chronic capsaicin stimulation. AB - Gamma-aminobutyric acid type B receptors (GABAB) are G-protein-coupled receptors that mediate GABAergic inhibition in the brain. Their functional expression is dependent upon the formation of heterodimers between GABAB1 and GABAB2 subunits, a process that occurs within the endoplasmic reticulum. However, the mechanisms that regulate GABAB receptor oligomerization at the plasma membrane remain largely unknown. We first characterized the functional cytoarchitecture of an organotypic co-culture model of rat dorsal root ganglia and spinal cord. Subsequently, we studied the interactions between GABAB subunits after chronic stimulation of sensory fibres with capsaicin. Surface labelling of recombinant proteins showed a decrease in subunit co-localization and GABAB2 labelling, after capsaicin treatment. In these conditions, fluorescence lifetime imaging measurements further demonstrated a loss of interactions between green fluorescent protein-GABAB1b and t-dimer discosoma sp red fluorescent protein GABAB2 subunits. Finally, we established that the GABAB receptor undergoes clathrin-dependent internalization and rapid recycling to the plasma membrane following activation with baclofen, a GABAB agonist. However, in cultures chronically stimulated with capsaicin, the agonist-induced endocytosis was decreased, reflecting changes in the dimeric state of the receptor. Taken together, our results indicate that the chronic stimulation of sensory fibres can dissociate the GABAB heterodimer and alters its responsiveness to the endogenous ligand. Chronic stimulation thus modulates receptor oligomerization, providing additional levels of control of signalling. PMID- 17425568 TI - Neural pathways associated with loss of consciousness caused by intracerebral microinjection of GABA A-active anesthetics. AB - Anesthesia, slow-wave sleep, syncope, concussion and reversible coma are behavioral states characterized by loss of consciousness, slow-wave cortical electroencephalogram, and motor and sensory suppression. We identified a focal area in the rat brainstem, the mesopontine tegmental anesthesia area (MPTA), at which microinjection of pentobarbital and other GABA(A) receptor (GABA(A)-R) agonists reversibly induced an anesthesia-like state. This effect was attenuated by local pre-treatment with the GABA(A)-R antagonist bicuculline. Using neuroanatomical tracing we identified four pathways ascending from the MPTA that are positioned to mediate electroencephalographic synchronization and loss of consciousness: (i) projections to the intralaminar thalamic nuclei that, in turn, project to the cortex; (ii) projections to several pontomesencephalic, diencephalic and basal forebrain nuclei that project cortically and are considered parts of an ascending "arousal system"; (iii) a projection to other parts of the subcortical forebrain, including the septal area, hypothalamus, zona incerta and striato-pallidal system, that may indirectly affect cortical arousal and hippocampal theta rhythm; and (iv) modest projections directly to the frontal cortex. Several of these areas have prominent reciprocal projections back to the MPTA, notably the zona incerta, lateral hypothalamus and frontal cortex. We hypothesize that barbiturate anesthetics and related agents microinjected into the MPTA enhance the inhibitory response of local GABA(A)-R-bearing neurons to endogenous GABA released at baseline during wakefulness. This modulates activity in one or more of the identified ascending neural pathways, ultimately leading to loss of consciousness. PMID- 17425569 TI - Physiological effects of auditory nerve myelinopathy in chinchillas. AB - The goals were to study the physiological effects of auditory nerve myelinopathy in chinchillas and to test the hypothesis that myelin abnormalities could account for auditory neuropathy, a hearing disorder characterized by absent auditory brainstem responses (ABRs) with preserved outer hair cell function. Doxorubicin, a cytotoxic drug used as an experimental demyelinating agent, was injected into the auditory nerve bundle of 18 chinchillas; six other chinchillas were injected with vehicle alone. Cochlear microphonics, compound action potentials (CAPs), inferior colliculus evoked potentials (IC-EVPs), cubic distortion product otoacoustic emissions and ABRs were recorded before and up to 2 months after injection. Cochleograms showed no hair cell loss in any of the animals and measures of outer hair cell function were normal (cubic distortion product otoacoustic emissions) or enhanced (cochlear microphonics) after injection. ABR was present in animals with mild myelin damage (n = 10) and absent in animals with severe myelin damage that included the myelin surrounding spiral ganglion cell bodies and fibers in Rosenthal's canal (n = 8). Animals with mild damage had reduced response amplitudes at 1 day, followed by recovery of CAP and enhancement of the IC-EVP. In animals with severe damage, CAP and IC-EVP thresholds were elevated, amplitudes were reduced, and latencies were prolonged at 1 day and thereafter. CAPs deteriorated over time, whereas IC-EVPs partially recovered; latencies remained consistently prolonged despite changes in amplitudes. The results support auditory nerve myelinopathy as a possible pathomechanism of auditory neuropathy but indicate that myelinopathy must be severe before physiological measures are affected. PMID- 17425570 TI - Changes in neuronal response to ischemia in retinas with genetic alterations of somatostatin receptor expression. AB - Ischemia is a primary cause of neuronal death in retinal diseases. The repertoire of expressed transmitter receptors would determine the neurons' responses to ischemic damage, and peptidergic receptors may be involved. With a new in vitro model of the ischemic mouse retina, we investigated whether an altered expression of somatostatin receptors could modulate retinal responses to ischemia. We used retinas of somatostatin receptor 1 (sst(1)) knock out (KO) mice, where sst(2) are over-expressed and over-functional, and of sst(2) KO mice. TUNEL analysis of ischemic retinas showed a marked reduction of cell death in sst(1) KO retinas, while there were no differences between wild-type (WT) and sst(2) KO retinas. In addition, caspase-3 mRNA expression was also reduced in sst(1) KO as compared to WT retinas. An immunohistochemical analysis demonstrated that different cell populations responded differently to the ischemic insult, and that the persistence of some immunohistochemical markers was greater in sst(1) KO than in WT or in sst(2) KO retinas. In particular, rod bipolar cell survival was markedly improved in sst(1) KO retinas, while it was dramatically decreased in sst(2) KO retinas. Furthermore, consistent with a role of glutamate excitotoxicity in ischemia-induced neuronal death, retinal glutamate release was observed to increase under ischemic conditions, but this increase was significantly reduced in sst(1) KO retinas. These observations demonstrate that an increased presence of functional sst(2) protects against retinal ischemia, thus implementing the background for the use of sst(2) analogs in therapies of retinal diseases such as glaucoma or diabetic retinopathy. PMID- 17425571 TI - The neuropeptide corticotropin-releasing factor regulates excitatory transmission and plasticity at the climbing fibre-Purkinje cell synapse. AB - The climbing fibre (CF) input controls cerebellar Purkinje cell (PC) activity as well as synaptic plasticity at parallel fibre (PF)-PC synapses. Under high activity conditions, CFs release not only glutamate, but also the neuropeptide corticotropin-releasing factor (CRF). Brief periods of such high CF activity can lead to the induction of long-term depression (LTD) at CF-PC synapses. Thus, we have examined for the first time the role of CRF in regulating excitatory postsynaptic currents (EPSCs) and long-term plasticity at this synapse. Exogenous application of CRF alone transiently mimicked three aspects of CF-LTD, causing reductions in the CF-evoked excitatory postsynaptic current, complex spike second component and complex spike afterhyperpolarization. The complex spike first component is unaffected by CF-LTD induction and was similarly unaffected by CRF. Application of a CRF receptor antagonist reduced the expression amplitude and induction probability of CF-LTD monitored at the EPSC level. Collectively, these results suggest that under particular sensorimotor conditions, co-release of CRF from climbing fibres could down-regulate excitatory transmission and facilitate LTD induction at CF-PC synapses. Inhibition of either protein kinase C (PKC) or protein kinase A (PKA) attenuated the effects of CRF upon CF-EPSCs. We have previously shown that CF-LTD induction is PKC-dependent, and here demonstrate PKA dependence as well. These results suggest that both the acute effects of CRF on CF-EPSCs as well as the facilitating effect of CRF on CF-LTD induction can be explained by a CRF-mediated recruitment of PKC and PKA. PMID- 17425572 TI - Increased occurrence of climbing fiber inputs to the cerebellar flocculus in a mutant mouse is correlated with the timing delay of optokinetic response. AB - The cerebellum plays an essential role in motor control, and its dysfunction may delay the onset of action and disrupt smooth and efficient movement. A Purkinje neuron (PN), the sole output cell type in the cerebellar cortex, receives two distinct types of excitatory synaptic inputs, numerous weak inputs from granule neurons (GNs) and occasional strong inputs from a climbing fiber (CF). The role of each input and the significance of low firing rate of CF have been studied. Here we show that the increased occurrence of CF inputs altered the firing pattern of a PN, which was correlated with timing of a reflex. We used the mutant mice deficient in the glutamate receptor delta2 subunit, a molecule related to ionotropic glutamate receptor specifically expressed at GN-PN synapses. The mutant mouse shows more frequent CF inputs and longer timing delay in optokinetic response (OKR), reflex eye movement that follows slow motion of a large visual field. A PN shows two types of action potentials: complex spikes (CS) induced by CF inputs; and simple spikes. They changed respective firing rates during sinusoidal optokinetic stimulation, and the timing of each firing rate modulation was similar in wild-type and mutant mice. However, increased occurrence of CS in the mutant altered the total firing pattern of a PN in the flocculus, which was correlated with the timing delay of OKR. These results support the functional merit of low firing rate of CF in motor control. PMID- 17425573 TI - Developmental neural plasticity and its cognitive benefits: olivocerebellar reinnervation compensates for spatial function in the cerebellum. AB - The adult mammalian central nervous system displays limited reinnervation and recovery from trauma. However, during development, post-lesion plasticity may generate alternative paths, thus providing models to investigate reinnervation and repair. After unilateral transection of the neonatal rat olivocerebellar path (pedunculotomy), axons from the remaining inferior olive reinnervate the denervated hemicerebellum. Unfortunately, reinnervation to the cerebellar hemisphere is incomplete; therefore, its capacity to mediate hemispheric function (navigation) is unknown. We studied sensorimotor control and spatial cognition of rats with and without transcommissural reinnervation using simple (bridge and ladder) and complex (wire) locomotion tests and the Morris water maze (hidden, probe and cued paradigms). Although pedunculotomized animals completed locomotory tasks more slowly than controls, all groups performed equally in the cued maze, indicating that lesioned animals could orientate to and reach the platform. In animals pedunculotomized on day 3 (Px3), which develop olivocerebellar reinnervation, final spatial knowledge was as good as controls, although they learned more erratically, failing to retain all information from one day to the next. By contrast, animals pedunculotomized on day 11 (Px11), which do not develop reinnervation, did not learn the task, taking less direct routes and more time to reach the platform than controls. In the probe test, control and Px3, but not Px11, animals swam directly to the remembered location. Furthermore, the amount of transcommissural reinnervation to the denervated hemisphere correlated directly with spatial performance. These results show that transcommissural olivocerebellar reinnervation is associated with spatial learning, i.e. even partial circuit repair confers significant functional benefit. PMID- 17425574 TI - Role of GABA A inhibition in modulation of pyramidal tract neuron activity during postural corrections. AB - In a previous study we demonstrated that the activity of pyramidal tract neurons (PTNs) of the motor cortex is modulated in relation to postural corrections evoked by periodical tilts of the animal. The modulation included an increase in activity in one phase of the tilt cycle and a decrease in the other phase. It is known that the motor cortex contains a large population of inhibitory GABAergic neurons. How do these neurons participate in periodic modulation of PTNs? The goal of this study was to investigate the role of GABA(A) inhibitory neurons of the motor cortex in the modulation of postural-related PTN activity. Using extracellular electrodes with attached micropipettes, we recorded the activity of PTNs in cats maintaining balance on a tilting platform both before and after iontophoretic application of the GABA(A) receptor antagonists gabazine or bicuculline. The tilt-related activity of 93% of PTNs was affected by GABA(A) receptor antagonists. In 88% of cells, peak activity increased by 75 +/- 50% (mean +/- SD). In contrast, the trough activity changed by a much smaller value and almost as many neurons showed a decrease as showed an increase. In 73% of the neurons, the phase position of the peak activity did not change or changed by no more than 0.1 of a cycle. We conclude that the GABAergic system of the motor cortex reduces the posture-related responses of PTNs but has little role in determining their response timing. PMID- 17425575 TI - Impact of chronic subthalamic high-frequency stimulation on metabolic basal ganglia activity: a 2-deoxyglucose uptake and cytochrome oxidase mRNA study in a macaque model of Parkinson's disease. AB - The mechanisms of action of high-frequency stimulation (HFS) of the subthalamic nucleus (STN) remain only partially understood. Hitherto, experimental studies have suggested that STN-HFS reduces the activity of STN neurons. However, some recent reports have challenged this view, showing that STN-HFS might also increase the activity of globus pallidus internalis (GPi) neurons that are under strong excitatory drive of the STN. In addition, most results emanate from studies applying acute STN-HFS, while parkinsonian patients receive chronic stimulation. Thus, the present study was designed to assess the effect of chronic (10 days) STN-HFS in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treated nonhuman primate. For this purpose, 2-deoxyglucose (2-DG) uptake, a measure of global synaptic activity, was assessed in the basal ganglia and the motor thalamus after chronic unilateral STN-HFS. Cytochrome oxidase subunit 1 (COI) mRNA expression, a marker of efferent metabolic activity, was additionally assessed in the globus pallidus. Chronic STN-HFS (i) reversed abnormally decreased 2-DG uptake in the STN of parkinsonian nonhuman primates, (ii) reversed abnormally increased 2-DG accumulation in the GPi while COI mRNA expression was increased, suggesting global activation of GPi neurons, and (iii) reversed abnormally increased 2-DG uptake in the ventrolateral motor thalamus nucleus. The simultaneous decrease in 2-DG uptake and increase in COI mRNA expression are difficult to reconcile with the current model of basal ganglia function and suggest that the mechanisms by which STN-HFS exerts its clinical benefits are more complex than a simple reversal of abnormal activity in the STN and its targets. PMID- 17425576 TI - Dorsolateral cervical spinal injury differentially affects forelimb and hindlimb action in rats. AB - In experimental spinal injury studies, damage to the dorsal half of the spinal cord is common but the behavioural effects of damage to specific pathways in the dorsal cord have been less well investigated. We performed bilateral transection of the dorsolateral spinal funiculus (DLF) on 12 Long-Evans rats at the third cervical spinal segment. We quantified overground locomotion by measuring ground reaction forces, step timing and step distances as animals moved unrestrained. We also assessed skilled locomotion by measuring footslip errors made while the animals crossed horizontal ladders, and examined paw usage in a cylinder exploration task and during a skilled reaching task. Ground reaction forces revealed that rats with bilateral DLF lesions moved with a symmetrical gait, characterized mainly by altered forces exerted by the hindlimbs, delayed onset of hindlimb stance, and understepping of the hindlimbs relative to the forelimbs. These alterations in overground locomotion were subtle but were nevertheless consistent between animals and persisted throughout the 6-week recovery period. During ladder crossing, rats with DLF lesions made more footslip errors with the hindlimbs after surgery than before. Spontaneous forelimb usage during exploration was not affected by DLF axotomy but lesioned animals were less successful during skilled reaching. This is the first study which describes preferentially altered hindlimb use during overground locomotion after cervical DLF transections. We discuss these findings in relation to previous work and to the possible contributions of different ascending and descending pathways in the DLF to locomotion and skilled movements in rats. PMID- 17425577 TI - Olfactory lateralization in homing pigeons: initial orientation of birds receiving a unilateral olfactory input. AB - It has been shown that homing pigeons (Columba livia) rely on olfactory cues to navigate from unfamiliar locations. In fact, the integrity of the olfactory system, from the olfactory mucosa to the piriform cortex, is required for pigeons to navigate over unfamiliar areas. Recently it has been shown that there is a functional asymmetry in the piriform cortex, with the left piriform cortex more involved in the use of the olfactory navigational map than the right piriform cortex. To investigate further the lateralization of the olfactory system in relation to navigational processes in carrier pigeons, we compared their homing performance after either their left or the right nostril was plugged. Contrary to our expectations, we observed an impairment in the initial orientation of the pigeons with their right nostril plugged. However, both groups released with one nostril plugged tended to be poorer than control pigeons in their homing performance. The observed asymmetry in favour of the right nostril might be due to projections from the olfactory bulbs to the contralateral globus pallidum, a structure involved in motor responses. PMID- 17425578 TI - Sex differences in catechol contents in the olfactory bulb of control and unilaterally deprived rats. AB - The dopaminergic system plays important roles in the modulation of olfactory transmission. The present study examines the distribution of dopaminergic cells and the content of dopamine (DA) and its metabolites in control and deprived olfactory bulbs (OB), focusing on the differences between sexes. The content of DA and of its metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were measured by HPLC. The morphology and distribution of dopaminergic neurons were studied using tyrosine hydroxylase (TH) immunohistochemistry. Cells were typified with TH-parvalbumin, TH-cholecystokinin or TH-neurocalcin double-immunofluorescence assays. Biochemical analyses revealed sex differences in the content of DA and of its metabolites. In normal conditions, the OBs of male rats had higher concentrations of DA, DOPAC and HVA than the OBs of females. The immunohistochemical data pointed to sex differences in the number of TH-immunopositive cells (higher in male than in female rats). Colocalization analyses revealed that dopaminergic cells constitute a different cell subpopulation from those labelled after parvalbumin, cholecystokinin or neurocalcin immunostaining. Unilateral olfactory deprivation caused dramatic alterations in the dopaminergic system. The DA content and the density of dopaminergic cells decreased, the contents of DA and DOPAC as well as TH immunoreactivity were similar in deprived males and females and, finally, the metabolite/neurotransmitter ratio increased. Our results show that the dopaminergic modulation of olfactory transmission seems to differ between males and females and that it is regulated by peripheral olfactory activity. A possible role of the dopaminergic system in the sexually different olfactory sensitivity, discrimination and memory is discussed. PMID- 17425579 TI - Seasonal variations of clock gene expression in the suprachiasmatic nuclei and pars tuberalis of the European hamster (Cricetus cricetus). AB - In mammals, day length (photoperiod) is read and encoded in the main circadian clock, the suprachiasmatic nuclei (SCN). In turn, the SCN control the seasonal rhythmicity of various physiological processes, in particular the secretion pattern of the pineal hormone melatonin. This hormone then operates as an essential mediator for the control of seasonal physiological functions on some tissues, especially the pars tuberalis (PT). In the European hamster, both hormonal (melatonin) and behavioral (locomotor activity) rhythms are strongly affected by season, making this species an interesting model to investigate the impact of the seasonal variations of the environment. The direct (on SCN) and indirect (via melatonin on PT) effect of natural short and long photoperiod was investigated on the daily expression of clock genes, these being expressed in both tissues. In the SCN, photoperiod altered the expression of all clock genes studied. In short photoperiod, whereas Clock mRNA levels were reduced, Bmal1 expression became arrhythmic, probably resulting in the observed dramatic reduction in the rhythm of Avp expression. In the PT, Per1 and Rev-erbalpha expressions were anchored to dawn in both photoperiods. The daily profiles of Cry1 mRNA were not concordant with the daily variations in plasma melatonin although we confirmed that Cry1 expression is regulated by an acute melatonin injection in the hamster PT. The putative role of such seasonal-dependent changes in clock gene expression on the control of seasonal functions is discussed. PMID- 17425580 TI - Implication of protein kinase C in the orexin-induced elevation of extracellular dopamine levels and its rewarding effect. AB - In the present study, we investigated the role of orexinergic systems in the activation of midbrain dopamine neurons. In an in vitro study, exposure to either orexin A or orexin B under superfusion conditions produced a transient increase in the intracellular Ca(2+) concentration through the phospholipase C (PLC)/protein kinase C (PKC) pathway via G(q11)alpha or Gbetagamma subunits in midbrain cultured neurons, which were shown to be tyrosine hydroxylase (TH) positive cells, but not in purified midbrain astrocytes. Here we show that in vivo injection with a selective PKC inhibitor chelerythrine chloride or 2-{8 [(dimethylamino)methyl]-6,7,8,9-tetrahydropyrido[1,2-a]indol-3-yl}-3-1-methyl-1H indol-3-ylmaleimide HCl (Ro-32-0432) into the ventral tegmental area (VTA) significantly suppressed the place preference and increased levels of dopamine in the nucleus accumbens (NAcc) induced by intra-VTA injection of orexins. These results strongly support the idea that activation of the orexin-containing neuron in the VTA leads to the direct activation of mesolimbic dopamine neurons through the activation of the PLC/PKC pathway via G(q11)alpha or Gbetagamma-subunit activation, which could be associated with the development of its rewarding effect. PMID- 17425581 TI - Impact of 6-hydroxydopamine lesions and cocaine exposure on mu-opioid receptor expression and regulation of cholinergic transmission in the limbic-prefrontal territory of the rat dorsal striatum. AB - Information processing within the striatum is regulated by local circuits involving dopamine, cholinergic interneurons and neuropeptides released by recurrent collaterals of striatal output neurons. In the limbic-prefrontal territory of the dorsal striatum, enkephalin inhibits the NMDA-evoked release of acetylcholine directly through micro-opioid receptors (MORs) located on cholinergic interneurons and indirectly through MORs of output neurons of striosomes. In this territory, we investigated the consequence of changes in dopamine transmission, bilateral 6-hydroxydopamine-induced degeneration of striatal dopaminergic innervation or cocaine (acute and chronic) exposure on (i) MOR expression in both cholinergic interneurons and output neurons of striosomes, and (ii) the direct and indirect enkephalin-MOR regulations of the NMDA-evoked release of acetylcholine. Expression of MORs in cholinergic interneurons was preserved after 6-hydroxydopamine and down-regulated after cocaine treatments. Accordingly, the direct enkephalin-MOR control of acetylcholine release was preserved after 6-hydroxydopamine treatment and lost after cocaine exposure. Expression of MORs in output neurons of striosomes was down-regulated in the 6 hydroxydopamine situation and either preserved or up-regulated after acute or chronic cocaine exposure, respectively. Accordingly, the indirect enkephalin-MOR control of acetylcholine release disappeared in the 6-hydroxydopamine situation but surprisingly, despite preservation of MORs in striosomes, disappeared after cocaine treatment. Showing that MORs of striosomes are still functional in this situation, the MOR agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin inhibited the NMDA-evoked release of acetylcholine after cocaine exposure. Therefore, alteration in the regulation of cholinergic transmission by the enkephalin-MOR system might play a major role in the motivational and cognitive disorders associated with dopamine dysfunctions in fronto-cortico-basal ganglia circuits. PMID- 17425583 TI - Empirical tests of the functional significance of amygdala-based modulation of hippocampal representations: evidence for multiple memory consolidation pathways. AB - This series of experiments evaluated the effects of amygdala damage on the acquisition and long-term retention of variants of the water task, and tested the hypothesis that the amygdala is an essential neural system for consolidation of hippocampal memories. In Experiment 1, rats with large, neurotoxic lesions of the amygdala (AMYG) showed normal acquisition on the standard spatial version of the water task, as well as normal retention and decay rate profiles on the 24-h and 30-day retention probes. In Experiment 2, AMYG rats showed normal one-trial place learning abilities and could retain this one-trial information over a 24 h delay. Experiment 3 showed that the amygdala lesions used in this study were functionally significant because AMYG rats, from Experiment 2, showed impairments in a discriminative fear conditioning to context paradigm. Experiment 4 was a critical test of the idea that the amygdala is a decisive locus for consolidation of hippocampal memories. AMYG rats were trained to sub-asymptotic levels of performance on the standard version of the water task. Following each training session, the subjects were given a post-training peripheral injection of D amphetamine. A probe test revealed that normal subjects and AMYG rats showed similar post-training memory improvement effects. Taken together, the results show that hippocampal memory consolidation processes do not require amygdala modulation. Arguments for an alternative view are presented suggesting that there are multiple memory consolidation pathways, one of which may depend on amygdala neural circuitry. PMID- 17425582 TI - Dissociable effects of disconnecting amygdala central nucleus from the ventral tegmental area or substantia nigra on learned orienting and incentive motivation. AB - Recent evidence suggests that the amygdala central nucleus (CeA) and midbrain striatal dopamine systems are critically involved in the alteration of attentional and emotional processing of initially neutral stimuli by associative learning. In rats, the acquisition of learned orienting responses (ORs) to visual cues paired with food is impaired by lesions of the CeA, and by lesions that disconnect CeA from the dorsolateral striatum (DLS), a region traditionally implicated in elevated responsiveness to sensory stimuli. Similarly, the acquisition of emotional significance to cues paired with food also depends on the function of CeA and of the ventral striatal nucleus accumbens (ACB), a region often considered crucial to acquired reward and motivation. For example, the ability of a cue previously paired with food to increase the rate of food reinforced instrumental responding (Pavlovian-instrumental transfer, PIT) is eliminated by lesions of the CeA or the accumbens core. In this experiment, we found that lesions that functionally disconnected CeA from the substantia nigra pars compacta (SNc) impaired the acquisition of conditioned orienting to auditory cues paired with food, but had no effect on their ability to enhance instrumental responding, relative to the effects of unilateral lesions of that region. By contrast, lesions that disconnected CeA from the ventral tegmental area (VTA) had no effect on the acquisition of conditioned orienting, but facilitated Pavlovian instrumental transfer relative to unilateral midbrain lesions, rescuing that function to sham-lesion control levels. Otherwise, unilateral lesions of either midbrain region impaired transfer. Implications of these results for circuit models of amygdalo-striatal interactions in associative learning are discussed. PMID- 17425584 TI - Impaired hippocampal LTP in inbred mouse strains can be rescued by beta adrenergic receptor activation. AB - Long-term potentiation (LTP), an activity-dependent enhancement of synaptic strength, and memory can be influenced by neuromodulatory transmitters such as norepinephrine (NE) and also by genetic background. beta-Adrenergic receptor activation can facilitate the expression of hippocampal CA1 LTP induced by weak stimulus patterns, but its influence on LTP induced by stronger stimulus patterns is unclear. We examined neural NE and dopamine (DA) levels, beta-adrenergic receptor expression and hippocampal LTP in genetically diverse inbred mouse strains. Brain tissue levels of NE were significantly lower in strains 129S1/SvImJ (129), BALB/cByJ (BALB) and C3H/HeJ (C3H) than in C57BL/6NCrlBR (B6). Western blot analysis showed that hippocampal beta(1)-adrenergic receptor expression was similar in strains B6, 129 and C3H, but was increased in BALB. LTP was induced in area CA1 of hippocampal slices by four trains of high-frequency stimulation (HFS) of the Schaeffer collaterals in the four inbred strains. Two hours after induction, LTP was significantly reduced in strains 129, BALB and C3H compared to B6, correlating with neural NE levels. We rescued hippocampal LTP in strains 129, BALB and C3H to levels seen in B6 by bath application of 1 microm isoproterenol, a beta-adrenergic receptor agonist, during HFS. Propranolol, a beta-adrenergic receptor antagonist, blocked this rescue in 129, BALB and C3H but did not affect LTP in strain B6. Thus, although this form of multitrain LTP does not rely on beta-adrenergic receptor activation, our data show that pharmacological activation of beta-adrenergic receptors during multiple trains of HFS can rescue CA1 LTP in genetically diverse strains with impaired LTP. PMID- 17425585 TI - Social stress induces hypothalamo-pituitary-adrenal axis responses in lactating rats bred for high trait anxiety. AB - Hypothalamo-pituitary-adrenal axis responses to various stressors are typically attenuated during lactation, including in rats selectively bred for high or low anxiety. As high-anxiety dams are more aggressive towards intruders than low anxiety dams during maternal defence, we investigated their hypothalamo-pituitary adrenal axis responses to this social stress. Maternal defence induced elevated stress responses in high-anxiety dams only; nerve-growth-factor-induced gene B mRNA expression in the parvocellular paraventricular nucleus and adrenocorticotropin hormone secretory responses were substantially enhanced after maternal defence. In contrast, secretory responses to a non-social stress (elevated platform) were not different between high- and low-anxiety dams. Thus, responsiveness of the stress axis in lactation is dependent upon the innate level of anxiety of the dam and, as a consequence, her reactiveness to social threat. PMID- 17425586 TI - Task-related differential dynamics of EEG alpha- and beta-band synchronization in cortico-basal motor structures. AB - Movement-related processing results in the modulation of neuronal synchronization over several electroencephalography (EEG) frequency ranges, including alpha- (8 12 Hz) and beta-band (14-30 Hz). Whether modulation patterns differ across sites within the motor system remains unclear, but could denote how information is conveyed across the cortico-basal network. We therefore compared the event related synchronization/desynchronization (ERS/ERD) in recordings from the scalp, basal ganglia and thalamic structures during a motor task. Simultaneous depth and scalp EEG were recorded in 13 patients, undergoing deep brain stimulation of the thalamic ventral intermediate nucleus (VIM) or the subthalamic nucleus (STN). They performed a choice-reaction task with pre-cued Go-signals, instructive for either left- or right-sided button presses. In the beta-band, pre-cues and Go signals were followed by ERD starting well before and peaking at task execution, uniformly in all cortical and subcortical recordings. In contrast, a comparable alpha-band ERD was only seen at the scalp, whereas mirror-like ERS were observed in the motor-inhibitory STN. In VIM, which receives strong somatosensory afferences, a major alpha-ERD upon the Go-signal did not start until the motor response. These dissociations of task-related Alpha- and Beta-band dynamics tag a functional diversity in cortico-basal networks, which are simultaneously active in motor processing. Whereas the uniform downregulation of Beta-activity points to an anti-kinetic operation mode throughout the motor system, site-dependent courses of Alpha-synchronization rather reflect the coordination of activity levels in functionally divergent motor structures during the preparation and execution of movements. PMID- 17425590 TI - Significance of error-avoiding mechanisms for oxidative DNA damage in carcinogenesis. AB - Reactive oxygen species (ROS) are produced through normal cellular metabolism, and their formation is further enhanced by exposure to ionizing radiation and various chemicals. ROS attack DNA, and the resulting oxidative DNA damage is considered to contribute to aging, carcinogenesis and neurodegeneration. Among various types of oxidative DNA damage, 8-oxo-7,8-dihydroguanine (8-oxoguanine or 8-oxoG) is the most abundant, and plays significant roles in mutagenesis because of its ability to pair with adenine as well as cytosine. Enzymatic activities that may be responsible for preventing 8-oxoG-evoked mutations were identified in mammalian cells. We have focused on the following three enzymes: MTH1, OGG1 and MUTYH. MTH1 is a mammalian ortholog of Escherichia coli MutT, which hydrolyzes 8 oxo-dGTP to its monophosphate form in nucleotide pools, thereby preventing incorporation of the mutagenic substrate into DNA. OGG1, a functional counterpart of E. coli MutM, has an 8-oxoG DNA glycosylase activity. MUTYH, a mammalian ortholog of E. coli MutY, excises an adenine paired with 8-oxoG. These three enzymes are thought to prevent mutagenesis caused by 8-oxoG in mammals. To analyze the functions of mammalian MTH1 (Mth1), OGG1 (Ogg1) and MUTYH (Mutyh) in vivo, we established mutant mice for these three enzymes by targeted mutagenesis, and investigated spontaneous tumorigenesis as well as mutagenesis. Here we discuss our recent investigation of mutagenesis and carcinogenesis in these mutant mice. PMID- 17425591 TI - Hypoxia increases the motility of lung adenocarcinoma cell line A549 via activation of the epidermal growth factor receptor pathway. AB - Tumor hypoxia is associated with a malignant phenotype of cancer cells and poor patient prognosis. To investigate the role of hypoxia in tumor progression, we studied the effects of hypoxia in the A549 lung adenocarcinoma cell line. First, we showed that hypoxic treatment decreased cell-cell adhesion and induced a scattering of cancer cells. Concomitant with these morphological changes, the motility of cancer cells was increased, as demonstrated by the Boyden chamber assay. Then, we used oligonucleotide array analyses to identify the genes causally related to the hypoxia-induced motile phenotype. The results showed that the expression of approximately 100 genes was induced more than 5-fold by hypoxia. These included (among others) epidermal growth factor receptor (EGFR), as well as other well-known hypoxia-induced genes, such as vascular endothelial growth factor. Immunohistochemical analyses of primary lung adenocarcinomas confirmed the induction of EGFR in tumor cells in the vicinity of necrotic areas, a histological indicator of tumor hypoxia. Remarkably, the EGFR inhibitor AG1478 (10 microM) completely blocked the increased cell motility induced by hypoxia. Thus, the present study demonstrates the importance of the EGFR pathway in the increased motility of cancer cells that occurs in a hypoxic tumor environment. PMID- 17425592 TI - Polymerase chain reaction-based fluorescent Luminex assay to detect the presence of human papillomavirus types. AB - Becuase 40% of human papillomavirus (HPV) infections are mixed infections, the accurate identification of high-risk HPV genotypes in mixed infections is important for defining a woman's risk for progression to cervical cancer. Thus, advanced Luminex-based HPV genotyping has been developed to simultaneously detect the presence of multiple HPV types. Here, we describe the development of a Luminex-based HPV genotyping that combines polymerase chain reaction amplification with hybridization to fluorescence-labeled polystyrene bead microarrays (Luminex suspension array technology). New HPV type-specific oligonucleotide probes and YBT L1/GP6-1 primers were used to detect the HPV types in 132 clinical samples. We simultaneously evaluated the usefulness of this technique on clinical samples. We detected 15 specific HPV types (6, 16, 18, 31, 35, 42, 51, 52, 55, 56, 58, 59, 66, 67 and 68) examined with specificity without known cross-reaction to other HPV types. The detection limit for the different HPV types was above 500 plasmids. We compared the performance of the Luminex based assay to the established HPV DNA microarray chip for polymerase chain reaction products derived from 53 clinical samples. The evaluation showed excellent agreement. The Luminex-based HPV genotyping was a sensitive, reproducible technique for the simultaneous genotyping of all clinically relevant genital HPV types. This assay system may be used to provide critical clinical information for early detection of HPV, especially in cases where the HPV copy numbers are low and the latency period of HPV infection is prolonged. PMID- 17425593 TI - Substrate choice of membrane-type 1 matrix metalloproteinase is dictated by tissue inhibitor of metalloproteinase-2 levels. AB - Although tissue inhibitor of metalloproteinase-2 (TIMP-2) is known to be not only an inhibitor of matrix metalloproteinases (MMP) but also a cofactor for membrane type 1 MMP (MT1-MMP)-mediated MMP-2 activation, it is still unclear how TIMP-2 regulates MMP-2 activation and cleavage of substrates by MT1-MMP. In the present study we examined the levels of cell-surface MT1-MMP, MMP-2 activation and cleavage of MT1-MMP substrates in 293T cells transfected with the MT1-MMP and TIMP-2 genes. Co-expression of TIMP-2 at an appropriate level increased the level of cell-surface MT1-MMP, both the TIMP-2-bound and free forms, and generated processed MMP-2 with gelatin-degrading activity. In contrast, MT1-MMP substrates testican-1 and syndecan-1 were cleaved by the cells expressing MT1-MMP, which was inhibited by TIMP-2 even at levels that stimulate MMP-2 activation. These results suggest that TIMP-2 environment determines MT1-MMP substrate choice between direct cleavage of its own substrates and MMP-2 activation. PMID- 17425594 TI - Genome-wide screening of loci associated with drug resistance to 5-fluorouracil based drugs. AB - Resistance to chemotherapeutic agents represents the chief cause of mortality in cancer patients with advanced disease. Chromosomal aberration and altered gene expression are the main genetic mechanisms of tumor chemoresistance. In this study, we have established an algorithm to calculate DNA copy number using the Affymetrix 10K array, and performed a genome-wide correlation analysis between DNA copy number and antitumor activity against 5-fluorouracil (5-FU)-based drugs (S-1, tegafur + uracil [UFT], 5'-DFUR and capecitabine) to screen for loci influencing drug resistance using 27 human cancer xenografts. A correlation analysis confirmed that the single nucleotide polymorphism (SNP) showing significant associations with drug sensitivity were concentrated in some cytogenetic regions (18p, 17p13.2, 17p12, 11q14.1, 11q11 and 11p11.12), and we identified some genes that have been indicated their relations to drug sensitivity. Among these regions, 18p11.32 at the location of the thymidylate synthase gene (TYMS) was strongly associated with resistance to 5-FU-based drugs. A change in copy number of the TYMS gene was reflected in the TYMS expression level, and showed a significant negative correlation with sensitivity against 5 FU-based drugs. These results suggest that amplification of the TYMS gene is associated with innate resistance, supporting the possibility that TYMS copy number might be a predictive marker of drug sensitivity to fluoropyrimidines. Further study is necessary to clarify the functional roles of other genes coded in significant cytogenetic regions. These promising data suggest that a comprehensive DNA copy number analysis might aid in the quest for optimal markers of drug response. PMID- 17425595 TI - Decrease in risk of lung cancer death in Japanese men after smoking cessation by age at quitting: pooled analysis of three large-scale cohort studies. AB - To evaluate the impact of smoking cessation on individuals and populations, we examined the decrease in risk of lung cancer death in male ex-smokers by age at quitting by pooling the data from three large-scale cohort studies in Japan. For simplicity, subjects were limited to male never smokers and former or current smokers who started smoking at ages 18-22 years, and 110,002 men aged 40-79 years at baseline were included. During the mean follow-up of 8.5 years, 968 men died from lung cancer. The mortality rate ratio compared to current smokers decreased with increasing attained age in men who stopped smoking before age 70 years. Among men who quit in their fifties, the cohort-adjusted mortality rate ratios (95% confidence interval) were 0.57 (0.40-0.82), 0.44 (0.29-0.66) and 0.36 (0.13 1.00) at attained ages 60-69, 70-79 and 80-89 years, respectively. The corresponding figures for those who quit in their sixties were 0.81 (0.44-1.48), 0.60 (0.43-0.82) and 0.43 (0.21-0.86). Overall, the mortality rate ratio for current smokers, relative to non-smokers, was 4.71 (95% confidence interval 3.76 5.89) and those for ex-smokers who had quit smoking 0-4, 5-9, 10-14, 15-19, 20-24 and >or=25 years before were 3.99 (2.97-5.35), 2.55 (1.80-3.62), 1.87 (1.23 2.85), 1.21 (0.66-2.22), 0.76 (0.33-1.75) and 0.67 (0.34-1.32), respectively. Although earlier cessation of smoking generally resulted in a lower rate of lung cancer mortality in each group of attained age, the absolute mortality rate decreased appreciably after stopping smoking even in men who quit at ages 60-69 years. PMID- 17425596 TI - Meat, fish and fat intake in relation to subsite-specific risk of colorectal cancer: The Fukuoka Colorectal Cancer Study. AB - High intake of red meat has been associated with increased risk of colorectal cancer in Western countries. There has been much interest in the role of n-3 polyunsaturated fatty acids (PUFA) in colorectal cancer prevention, but epidemiological findings are limited and inconsistent. The objective of our study was to examine associations of meat, fish and fat intake with risk of colorectal cancer, paying particular attention to the subsite within the colorectum. Data were from the Fukuoka Colorectal Cancer Study, a population-based case-control study, covering 782 cases and 793 controls. Diet was assessed by interview, using newly developed personal-computer software for registering semiquantitative food frequencies. The intake of beef/pork, processed meat, total fat, saturated fat or n-6 PUFA showed no clear association with the overall or subsite-specific risk of colorectal cancer. There was an almost significant inverse association between n 3 PUFA and the risk of colorectal cancer; the covariate-adjusted odds ratio for the highest (median 3.94 g/day) versus lowest (median 1.99 g/day) quintile of energy-adjusted intake was 0.74 (95% confidence interval 0.52-1.06, trend P=0.050). The consumption of fish and fish products was similarly inversely related to the risk although the association was not statistically significant. These associations were more evident for distal colon cancer; adjusted odds ratio for the highest versus lowest quintile of n-3 PUFA intake was 0.56 (95% confidence interval 0.34-0.92, trend P=0.02). Our findings do not support the hypothesis that consumption of red meat increases colorectal cancer risk but do suggest that high intake of fish may decrease the risk, particularly of distal colon cancer. PMID- 17425598 TI - Specialist nurses for older people: implications from UK development sites. AB - AIM: This paper is a report of a study to explore the development of specialist staffing for older people in six case study sites in the United Kingdom. BACKGROUND: In the United Kingdom there has been some concern about the health care available to older people, leading to the development of a National Service Framework. A key theme of this framework is the development of specialist staff skilled in providing services tailored to the needs of older people. METHOD: A soft systems methodology was used in 2004-2005 to carry out interviews with key people, including specialist nurses for older people, other service providers, patients and informal carers (n = 132) in six case study sites identified from a national questionnaire. Interviewees were asked to describe their perceptions of the development, its history and its impact. FINDINGS: The development of specialist nursing services seemed to be shaped by national policy drivers for service development, which may not have been directly linked to the needs of older people. The ideal qualities of a specialist nurse for older people were described by participants as including not only knowledge and skills, but also personal characteristics. CONCLUSION: While progress has been made in establishing specialist posts, much remains to be explored about the roles of postholders, the qualities needed, and the support and preparation required. While advanced practice is a professional aspiration, a number of questions arise about the development of nursing as a self-directing profession in diverse international settings. Theories of specialist nursing practice also need to address the tensions between universal and local models and to consider theories about nursing older people. PMID- 17425599 TI - Knowledge about social networks and integration: a co-operative research project. AB - AIM: This paper is a report of a study investigating how knowledge of social network and integration influenced mental health professionals' understanding and practice. BACKGROUND: Community mental health work focuses on people suffering from mental health problems as well as the consequences for the person involved and their family or network. There is a need to expand community mental health workers' knowledge about social networks and their functions. METHOD: A qualitative study using a co-operative research approach was used to develop participants' knowledge of social network and social integration theory. Action research has the potential to facilitate changes in the field. Data were collected using focus groups. Qualitative content analysis was employed to develop the theme and categories. The data were collected in 2004-2005. FINDINGS: The main theme identified was the potential of experiential knowledge-based competence, which was characterized by the following categories: (1) increased knowledge, (2) awareness of social interactions, (3) cross-disciplinary professionalism and (4) potential for changes in practice. Participants' knowledge and awareness of the potential of social integration as a tool for social network interventions were considerably strengthened. However, this knowledge needs to be implemented in practice. CONCLUSION: Co-operative research is an approach that can be beneficial in the public sector. To achieve the best possible results, the whole team must be involved and play an active part in all areas of the research project. If the groups involved are too large, participants' level of engagement may suffer. PMID- 17425600 TI - Lamentation and loss: expressions of caring by contemporary surgical nurses. AB - AIM: This paper is a report of a phenomenological study of caring from the perspective of nurses working on surgical wards. BACKGROUND: While care and caring are complex foundational nursing concepts which have received considerable and ongoing attention from theorists, researchers and clinicians, there has been little research into caring on surgical units. METHOD: A convenience sample of ten nurses working on surgical units in a public teaching hospital in Canada was interviewed using van Manen's phenomenological approach. Data were collected during 2001 using semi-structured interviews. FINDINGS: The major theme of lamentation and loss was identified from the data. Participants revealed a dichotomous tension between what caring should be and what actually occurs. This tension was pervasive and generated lament - an expression of grief and mourning for the loss of caring. The essential structures supporting this theme included lack of time, lack of caring support, tasking, increased acuity, lack of continuity of care, emotional divestment and not caring for each other. Loss and sadness were articulated and participants lamented and grieved about the loss of care in contemporary practice. CONCLUSION: The forces and influences described by participants undermined caring in the new practice milieu. If this is a glimpse of the future, then the values of the nursing profession may be under siege. Caring as the central core, the essence or unifying concept of nursing may be subject to marginalization in contemporary practice. PMID- 17425601 TI - Peroxisome proliferator-activated receptor-gamma agonists inhibit respiratory syncytial virus-induced expression of intercellular adhesion molecule-1 in human lung epithelial cells. AB - Respiratory syncytial virus (RSV) is the major causative agent of severe lower respiratory tract disease and death in infants worldwide. The epithelial cells of the airways are the target cells for RSV infection and the site of the majority of the inflammation associated with the disease. However, despite five decades of intensive RSV research there exist neither an effective active vaccine nor a promising antiviral and anti-inflammatory therapy. Recently, peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, has been shown to possess anti-inflammatory properties. Therefore, we hypothesized whether the detrimental increase of intercellular adhesion molecule-1 (ICAM-1) on RSV-infected lung epithelial cells (A549 and primary normal human bronchial epithelial cells (NHBE)) might be modulated by natural and synthetic PPAR-gamma agonists (15d-PGJ2, ciglitazone, troglitazone, Fmoc-Leu). Our data show that all PPAR-gamma agonists under study significantly down-regulated the RSV-induced expression of ICAM-1 on A549- and NHBE cells in a dose-dependent manner resulting in a reduced beta2 integrin mediated adhesion of monocytic effector cells (U937) to RSV-infected A549 cell monolayers. In contrast, the PPAR-alpha agonist bezafibrate had no impact on the RSV-induced ICAM-1 expression. The reduced ICAM-1 expression was associated with a diminished ICAM-1 mRNA level and binding activity of nuclear factor-kappaB (p65/p50) in A549 cells. These findings suggest that PPARgamma agonists have beneficial effects in the suppression of the inflammatory response during RSV infection and therefore might have clinical efficacy in the course of severe RSV infection. PMID- 17425602 TI - Bone morphogenetic protein-2/4 signalling pathway components are expressed in the human thymus and inhibit early T-cell development. AB - T-cell differentiation is driven by a complex network of signals mainly derived from the thymic epithelium. In this study we demonstrate in the human thymus that cortical epithelial cells produce bone morphogenetic protein 2 (BMP2) and BMP4 and that both thymocytes and thymic epithelium express all the molecular machinery required for a response to these proteins. BMP receptors, BMPRIA and BMPRII, are mainly expressed by cortical thymocytes while BMPRIB is expressed in the majority of the human thymocytes. Some thymic epithelial cells from cortical and medullary areas express BMP receptors, being also cell targets for in vivo BMP2/4 signalling. The treatment with BMP4 of chimeric human-mouse fetal thymic organ cultures seeded with CD34+ human thymic progenitors results in reduced cell recovery and inhibition of the differentiation of human thymocytes from CD4- CD8- to CD4+ CD8+ cell stages. These results support a role for BMP2/4 signalling in human T-cell differentiation. PMID- 17425603 TI - Expression of c-kit and Sca-1 and their relationship with multidrug resistance protein 1 in mouse bone marrow mononuclear cells. AB - P-glycoprotein (Pgp) and multidrug resistance protein 1 (MRP1) are members of the ATP-binding cassette (ABC) family of transporter proteins. Both molecules are membrane-associated, energy-dependent efflux pumps with different substrate selectivity and they may play a role in the activation, differentiation and function of haematopoietic cells. Mouse haematopoietic cells are characterized by the expression of the cell surface molecules c-kit and Sca-1. Herein, the presence and activities of Pgp and MRP1 in mouse bone marrow mononuclear cells (BMMC) and their relationship with the proteins c-kit and Sca-1 were evaluated. Pgp and MRP activities were measured based on the extrusion of rhodamine 123 (for Pgp) and Fluo-3 (for MRP). Cell populations were assessed by cytometry using anti c-kit and anti-Sca1 antibodies. Pgp activity was present in 5% of BMMC while 50% of BMMC cells showed MRP activity. These findings agreed with the proportion of cells expressing the MRP1 surface molecule (51.3 +/- 4.17%). About 14% of BMMC were positive for c-kit and/or Sca-1 (9.3% c-kit- Sca-1+, 4.2% c-kit+ Sca-1- and 0.9% c-kit+ Sca-1+). Among these subpopulations only c-kit- Sca-1+ cells presented Pgp activity (21.36%). On the other hand, MRP activity was present in all three subpopulations. Most cells (82.5%) of the c-kit+ Sca-1- subpopulation presented MRP1 activity compared to only 54.1% of c-kit+ Sca-1+ and 38.8% of c kit- Sca-1+. This study demonstrates the expression and activity of MRP1 in BMMC. While only a small proportion of precursor cells had Pgp activity, MRP1 activity was present among different subpopulations of precursor cells. Further studies are necessary to establish the role of these transporters in haematopoietic cells. PMID- 17425604 TI - CD27 expression discriminates between regulatory and non-regulatory cells after expansion of human peripheral blood CD4+ CD25+ cells. AB - It is clear that regulatory T cells (Treg) have an important role in preventing autoimmunity and modulating responses to pathogens. Full characterization of Treg cell function in human patients would be greatly facilitated by practical methods for expanding Treg in vitro. Methods for expansion have been reported but whether expression of surface and intracellular markers associated with freshly isolated Treg following expansion correlates with the maintenance of function is unclear. Our aim was to investigate the various methods of expansion and to correlate regulatory activity with expression of these markers. We show that, of the markers associated with freshly isolated Treg, only CD27 expression correlated with regulatory activity and could be used to isolate cells with regulatory activity from lines expanded from CD4+ CD25+ cells. Also, cells expressing high levels of the transcription factor forkhead box P3 (Foxp3) were confined to the CD27+ population within these lines. Expression of CD27 by cells in lines expanded from CD4+ CD25- cells varied depending on the stimulus used for expansion, but these lines did not have significant regulatory activity even when the CD27+ cells were tested. Analysis of synovial CD4+ CD25+ cells from reactive arthritis patients revealed that they were predominantly CD27 positive. This also applied to CD25(high) and CD25(intermediate) CD4+ cells, despite their reported different abilities to regulate. We conclude that, whilst CD27 is useful for identifying Treg in the cell lines obtained after expansion of CD4+ CD25+ cells, its expression may not reliably identify the Treg cell population in other T-cell populations such as those found in joints. PMID- 17425606 TI - The neuropeptide pituitary adenylate cyclase-activating polypeptide exerts anti apoptotic and differentiating effects during neurogenesis: focus on cerebellar granule neurones and embryonic stem cells. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) was originally isolated from ovine hypothalamus on the basis of its hypophysiotrophic activity. It has subsequently been shown that PACAP and its receptors are widely distributed in the central nervous system of adult mammals, indicating that PACAP may act as a neurotransmitter and/or neuromodulator. It has also been found that PACAP and its receptors are expressed in germinative neuroepithelia, suggesting that PACAP could be involved in neurogenesis. There is now compelling evidence that PACAP exerts neurotrophic activities in the developing cerebellum and in embryonic stem (ES) cells. In particular, the presence of PACAP receptors has been demonstrated in the granule layer of the immature cerebellar cortex, and PACAP has been shown to promote survival, inhibit migration and activate neurite outgrowth of granule cell precursors. In cerebellar neuroblasts, PACAP is a potent inhibitor of the mitochondrial apoptotic pathway through activation of the MAPkinase extracellular regulated kinase. ES cells and embryoid bodies (EB) also express PACAP receptors and PACAP facilitates neuronal orientation and induces the appearance of an electrophysiological activity. Taken together, the anti apoptotic and pro-differentiating effects of PACAP characterised in cerebellar neuroblasts as well as ES and EB cells indicate that PACAP acts not only as a neurohormone and a neurotransmitter, but also as a growth factor. PMID- 17425607 TI - Exposure to oestrogen prenatally does not interfere with the normal female typical development of odour preferences. AB - The neural mechanisms controlling mate recognition and heterosexual partner preference are sexually differentiated by perinatal actions of sex steroid hormones. We previously showed that the most important action of oestrogen during prenatal development is to defeminise and, to some extent, masculinise brain and behaviour in mice. Female mice deficient in alpha-foetoprotein (AFP) due to a targeted mutation in the Afp gene (AFP-KO) do not show any female sexual behaviour when paired with an active male because they lack the protective action of AFP against maternal oestrogens. In the present study, we investigated whether odour preferences, another sexually differentiated trait in mice, are also defeminised and/or masculinised in AFP-KO females due to their prenatal exposure to oestrogens. AFP-KO females of two background strains (CD1 and C57Bl/6j) preferred to investigate male over female odours when given the choice between these two odour stimuli in a Y-maze, and thus remained very female-like in this regard. Thus, the absence of lordosis behaviour in these females cannot be explained by a reduced motivation of AFP-KO females to investigate male-derived odours. Furthermore, the presence of a strong male-directed odour preference in AFP-KO females suggests a postnatal contribution of oestrogens to the development of preferences to investigate opposite-sex odours. PMID- 17425608 TI - Insulin-like growth factor-I mRNA and peptide in the human anterior pituitary. AB - The pituitary is the central organ regulating virtually all endocrine processes, and pathologies of the pituitary cause manifold adverse effects. Because insulin like growth factor (IGF)-I appears to be involved in tumour pathogenesis, progression, and persistence, and only few data exist on the cellular synthesis sites of IGF-I, the present study aims to create a basis for further research on pituitary adenomas by investigating the presence of IGF-I in the human pituitary using reverse transcriptase-polymerase chain reaction, in situ hybridisation, immunohistochemistry and immunocytochemistry. IGF-I was expressed in the pituitary, and gene sequence analysis revealed a sequence identical to that found in human liver. The distribution pattern of IGF-I mRNA found by in situ hybridisation corresponded to that of IGF-I peptide in immunohistochemistry. In all pituitary samples investigated, IGF-I-immunoreactivity occurred in almost all adrenocorticotrophic hormone (ACTH)-immunoreactive cells. Occasionally, an interindividually varying number of growth hormone (GH) and, infrequently, follicle-stimulating hormone and luteinising hormone cells contained IGF-I immunoreactivity but none was detected in supporting cells. At the ultrastructural level, IGF-I-immunoreactivity was confined to secretory granules in coexistence with ACTH- or GH-immunoreactivity, respectively, indicating a concomitant release of the hormones. Thus, in humans, IGF-I appears to be a constituent in ACTH cells whereas its production in GH-producing and gonadotrophic cells may depend on the physiological status (e.g. serum IGF-I level, age or reproductive phase). It is assumed that locally produced IGF-I plays a crucial role in the regulation of endocrine cells by autocrine/paracrine mechanisms in addition to the endocrine route. PMID- 17425609 TI - Gonadotrophin-releasing hormone (GnRH) release in marmosets I: in vivo measurement in ovary-intact and ovariectomised females. AB - In vivo hypothalamic gonadotrophin-releasing hormone (GnRH) release was characterised for the first time in a New World primate. A nonterminal and repeatable push-pull perfusion (PPP) technique reliably measured GnRH in conscious common marmoset monkeys. Nineteen adult females (n = 8 ovary-intact in the mid-follicular phase; n = 11 ovariectomised) were fitted with long-term cranial pedestals, and a push-pull cannula was temporarily placed in unique locations within the pituitary stalk-median eminence (S-ME) 2 days prior to each PPP session. Marmosets underwent 1-3 PPPs (32 PPPs in total) lasting up to 12 h. Plasma cortisol levels were not elevated in these habituated marmosets during PPP, and PPP did not disrupt ovulatory cyclicity or subsequent fertility in ovary intact females. GnRH displayed an organised pattern of release, with pulses occurring every 50.0 +/- 2.6 min and lasting 25.4 +/- 1.3 min. GnRH pulse frequency was consistent within individual marmosets across multiple PPPs. GnRH mean concentration, baseline concentration and pulse amplitude varied predictably with anatomical location of the cannula tip within the S-ME. GnRH release increased characteristically in response to a norepinephrine infusion and decreased abruptly during the evening transition to lights off. Ovary-intact (mid follicular phase) and ovariectomised marmosets did not differ significantly on any parameter of GnRH release. Overall, these results indicate that PPP can be used to reliably assess in vivo GnRH release in marmosets and will be a useful tool for future studies of reproductive neuroendocrinology in this small primate. PMID- 17425610 TI - Gonadotrophin-releasing hormone (GnRH) release in marmosets II: pulsatile release of GnRH and pituitary gonadotrophin in adult females. AB - Unlike other mammals, including rodents, Old World primates and humans, common marmosets and probably all other New World primates synthesise and release chorionic gonadotrophin (CG), and not luteinising hormone (LH) from pituitary gonadotrophs. However, little is known about the physiological dynamics of gonadotrophin-releasing hormone (GnRH)-regulated CG release from gonadotrophs and whether such CG release has pulsatile release characteristics similar to those of LH in other mammalian species. Consequently, we performed a series of in vivo and in vitro studies in ovariectomised laboratory rats and female marmosets to compare GnRH-induced pituitary LH and CG release characteristics, respectively. Exogenous GnRH stimulated a slower onset of release of marmoset pituitary CG, both in vivo and in vitro, and induced an approximately 400% greater increase in the duration of marmoset pituitary CG release compared to that for rat LH. Not surprisingly, hypothalamic pulsatile release of GnRH in vivo was not obviously concordant with endogenous episodic changes in circulating levels of CG in marmosets, in contrast to the clear concordance observed between in vivo GnRH and LH release previously demonstrated in rats and other mammals. Pituitary CG release in marmosets thus demonstrates considerable divergence from the timely hypothalamic GnRH-regulated LH release in other female mammals, implying potentially different physiological dynamics in gonadotrophin regulation of marmoset ovarian function. PMID- 17425611 TI - Various dietary fats differentially change the gene expression of neuropeptides involved in body weight regulation in rats. AB - Various high-fat diets are obesogenic but not to the same extent. The aim of the present study was to investigate the effects of saturated fat n-6 and n-3 polyunsaturated fatty acids (PUFAs) on the central neuropeptidergic system in adult rats. Using reverse transcriptase-polymerase chain reaction and in situ hybridisation, we evaluated the net effect of feeding in these fats, comparing the effects of a high- to low-fat diet, and the diversity of the effects of these fats in the same amount within the diet. We also determined plasma lipids, glucose, insulin and leptin concentrations. Six-week feeding with high-saturated fat evoked hyperpahagia and the largest weight gain compared to both high-PUFA diets. Rats fed high-saturated fat were found to have decreased neuropeptide Y (NPY) mRNA expression in the arcuate nucleus (ARC) and the compact zone of the dorsomedial nucleus (DMHc), unchanged pro-opiomelanocortin (POMC), galanin-like peptide (GALP) mRNA expression in the ARC, as well as melanin-concentrating hormone (MCH) and prepro-orexin (preORX) mRNA expression in the lateral hypothalamus, compared to low-saturated fed rats. By contrast, feeding with both high-PUFA diets increased POMC and GALP mRNA expression in the ARC compared to the corresponding low-fat diet and the high-saturated fat diet. Furthermore, feeding with both low-PUFA diets reduced NPY mRNA expression compared to the low saturated fat diet exclusively in the DMHc. Uniquely, the high n-3 PUFA feeding halved MCH and preORX mRNA expression in the lateral hypothalamus compared to the other high-fat and low n-3 PUFA diets. In rats fed three high-fat diets, plasma insulin and leptin concentrations were significantly increased and the type of fat had no effect on these hormone levels. Rats fed high-saturated fat had both hyperglycaemia and hypertriacylglycerolemia and rats fed high n-3 PUFA only had hyperglycaemia. The present study demonstrates that various forms of dietary fat differentially change the expression of neuropeptide genes involved in energy homeostasis. PMID- 17425612 TI - Noradrenaline release in the medial preoptic area during the rat oestrous cycle: temporal relationship with plasma secretory surges of prolactin and luteinising hormone. AB - During the rat oestrous cycle, the afternoon of pro-oestrous is characterised by preovulatory surges of luteinising hormone (LH) and prolactin. On the afternoon of oestrous, a secretory surge of prolactin has also been reported. Because the medial preoptic area (MPOA) is known to regulate prolactin and LH secretory surges and noradrenaline has been demonstrated to stimulate these hormones release, we evaluated whether noradrenaline release in the MPOA was temporally associated with plasma prolactin and LH surges in cycling rats. During the 4 days of oestrous cycle, noradrenaline concentrations were determined in microdialysates from the MPOA, collected at 30-min intervals from 10.30 h to 19.00 h. Plasma prolactin and LH levels were measured in blood samples withdrawn hourly from 14.00 h to 19.00 h on pro-oestrous and from 13.00 h to 18.00 h on the other days of the cycle. On the afternoons of both pro-oestrous and oestrous, noradrenaline levels increased at 14.00 h and remained elevated until 16.30 h. Conversely, they were low and constant throughout metoestrous and dioestrous. Correlating with noradrenaline release in the MPOA, plasma prolactin surges occurred during the afternoons of both pro-oestrous and oestrous. On pro oestrous, the afternoon LH surge was also preceded by the increase in MPOA noradrenaline whereas, during oestrous, LH secretion was low and unaltered. A temporal association between noradrenaline release and prolactin secretion suggests that noradrenergic neurotransmission in the MPOA regulates prolactin surges in female rats. Moreover, our data also suggest that MPOA noradrenaline requires specific conditions to physiologically regulate LH secretion, which seems to occur during the afternoon of pro-oestrous. PMID- 17425613 TI - Exogenous androgen treatment delays the stress response following hamster facial nerve injury. AB - Following injury or stress of any type, cells undergo a stress response, involving the cessation of general protein synthesis and the up-regulation of heat shock proteins (HSP), which have been implicated in promoting cell survival and repair. In a variety of neuronal injury models, including the hamster facial motoneurone (FMN) model, steroid hormones augment regeneration and are neuroprotective. We have previously shown that facial nerve axotomy induces expression of HSP70 (HSP70) and/or up-regulates constitutively expressed HSP70 (HSC70) mRNA in axotomised hamster FMN and that testosterone propionate (TP) treatment reduces this response. These previous studies were unable to differentiate between HSC70 mRNA and HSP70 mRNA. Therefore, an objective of the present study was to determine which HSP (HSC70 or HSP70) was being up-regulated by axotomy and reduced by TP. Axotomy increased HSC70 protein in axotomised and non-axotomised FMN, relative to untreated baseline hamsters. Interestingly, TP transiently delayed the stress-induced up-regulation of HSC70 protein in axotomised FMN compared to axotomised FMN from non-TP treated controls. A potential explanation for this delay may involve the TP-induced liberation of HSP from the androgen receptor, which would provide the injured cell with an immediately available pool of protective HSP. An hypothesis is presented suggesting that this TP-induced delay of stress-induced HSC70 up-regulation might allow for the diversion of cellular energy away from HSP synthesis and towards the synthesis of proteins required for regeneration and survival. PMID- 17425614 TI - The effects of GABA agonists on glutamic acid decarboxylase, GABA-transaminase, activin, salmon gonadotrophin-releasing hormone and tyrosine hydroxylase mRNA in the goldfish (Carassius auratus) neuroendocrine brain. AB - GABA plays a pivotal role in reproduction by regulating luteinising hormone (LH) release from the anterior pituitary. Current evidence indicates that there is a prominent stimulatory effect of GABA on LH release in teleost fish which results from enhanced gonadotrophin-releasing hormone (GnRH) release and decreased dopamine turnover in the brain and pituitary. We hypothesised that there may be additional mechanisms underlying LH release in goldfish and investigated the relative mRNA levels of GABA synthesising enzymes (GAD65 and GAD67), degrading enzyme (GABA-T), activin betaa and betab, salmon GnRH (sGnRH), and tyrosine hydroxylase (TH) with the real-time reverse transcriptase-polymerase chain reaction after GABA agonist treatment. Sexually regressed female goldfish were i.p. injected with either the GABA(A) agonist muscimol (1 microg/g body weight) or the GABA(B) agonist baclofen (10 microg/g body weight). Both agonists significantly increased serum LH after 6 h. Muscimol decreased GAD65 (approximately ten-fold), GABA-T (approximately 15-fold) and TH (approximately three-fold) mRNA in the telencephalon. Baclofen significantly reduced GAD67 (approximately two-fold) and GABA-T (approximately two-fold) mRNA levels in the hypothalamus. Activin betaa, but not activin betab, steady-state mRNA was increased approximately three- to four-fold in both the hypothalamus and telencephalon after baclofen treatment. There was no change in sGnRH mRNA levels in either tissue after GABA agonist treatment. We show that the GABA(A) and GABA(B) receptor agonists have differing and rapid effects on gene transcription in the goldfish neuroendocrine brain and, by affecting specific targets, we identify putative genomic mechanisms underlying GABA-stimulated LH release in fish. PMID- 17425615 TI - Intracardiac entrapment of a pulmonary artery catheter. PMID- 17425616 TI - Tramadol does not prolong the effect of ropivacaine 7.5 mg/ml for axillary brachial plexus block. AB - BACKGROUND: The aim of this prospective, randomized, double-blind study was to evaluate the effect of the addition of tramadol to ropivacaine on the onset and duration of sensory and motor block, and duration of analgesia, for axillary brachial plexus block. METHODS: After institutional approval and informed consent had been obtained, 45 patients scheduled for forearm or hand surgery under axillary brachial plexus block were randomly allocated into two groups. The ropivacaine group received 40 ml of ropivacaine 7.5 mg/ml plus 2 ml of isotonic sodium chloride solution, and the tramadol group received 40 ml of ropivacaine 7.5 mg/ml plus 2 ml (100 mg) of tramadol. The onset and duration of sensory and motor block in the distribution of the musculocutaneous, radial, median and ulnar nerves, the duration of analgesia, the time to first pain medication, hemodynamics and side-effects were recorded. RESULTS: The addition of tramadol did not improve the speed of onset or increase the duration of sensory and motor block. The durations of analgesia were 631 +/- 33 min and 633 +/- 37 min (mean +/ standard deviation) in the ropivacaine and tramadol groups, respectively (P > 0.05). Hemodynamic parameters and side-effects did not differ between the groups. CONCLUSION: The addition of 100 mg of tramadol to 7.5 mg/ml of ropivacaine, for axillary brachial plexus block, does not prolong the duration of motor and sensory block and analgesia. PMID- 17425617 TI - Increasing compression depth during manikin CPR using a simple backboard. AB - BACKGROUND: The quality of external chest compressions (ECC) is influenced by the surface supporting the patient. The aim of this study was to compare chest compression depth with and without a rigid backboard. The authors hypothesized that the presence of a backboard would result in an increased depth of chest compressions. METHODS: A randomized, double-blinded, cross-over trial. We simulated in-hospital cardiac arrest using a resuscitation manikin placed in a standard hospital bed. In total, 23 hospital orderlies were randomly assigned to perform ECC for 2 min on two identical ResusciAnne manikins, under one of which a rigid backboard had been placed. Data were recorded using the Laerdal PC-Skill Reporting System. RESULTS: Mean chest compression depth increased from 43 to 48 mm (P < 0.0001) when a backboard was present (mean difference 5 mm, 95% CI 3.6 7.5 mm, SD 4.6). There was a significant increase in mean proportion of compressions >40 mm when using a backboard Mean 92% vs. 69%, P= 0.0007). No difference was found between the two groups in the following variables: duty cycle, compression rate, mean proportion of compressions of correct depth (40-50 mm) or proportion of compressions with incomplete release. CONCLUSIONS: Applying a backboard significantly increases depth of chest compressions during cardiopulmonary resuscitation when performed on a manikin model. PMID- 17425618 TI - Early treatment with arteriovenous extracorporeal lung assist and high-frequency oscillatory ventilation in a case of severe acute respiratory distress syndrome. AB - BACKGROUND: Lung protective ventilation can reduce mortality in acute respiratory distress syndrome (ARDS). However, many patients with severe ARDS remain hypoxemic and more aggressive ventilation is necessary to maintain sufficient gas exchange. Pumpless arteriovenous extracorporeal lung assist (av-ECLA) has been shown to remove up to 95% of the systemic CO(2) production, thereby allowing ventilator settings and modes prioritizing oxygenation and lung protection. High frequency oscillatory ventilation (HFOV) is an alternative form of ventilation that may improve oxygenation while limiting the risk of further lung injury by using extremely small tidal volumes (VT). METHODS: We discuss the management of a patient suffering from severe ARDS as a result of severe bilateral lung contusions and pulmonary aspiration. RESULTS: Severe ARDS developed within 4 h after intensive care unit admission. Conventional mechanical ventilation (CV) with high-airway pressures and low VT failed to improve gas exchange. Av-ECLA was initiated to achieve a less aggressive ventilation strategy. VT was reduced to 2 3 ml/kg, but oxygenation did not improve and airway pressures remained high. HFOV (8-10 Hz) was started using a recruitment strategy and oxygenation improved within 2 h. After 5 days, the patient was switched back to CV uneventfully and av ECLA was removed after 8 days. CONCLUSION: The combination of two innovative treatment modalities resulted in rapid stabilization and improvement of gas exchange during severe ARDS refractory to conventional lung protective ventilation. During av-ECLA, extremely high oscillatory frequencies were used minimizing the risk of baro- and volutrauma. PMID- 17425619 TI - The epidural space is deeper in elderly and obese patients in the Japanese population. AB - BACKGROUND: Epidural anaesthesia is an efficient method of providing both regional anaesthesia and post-operative pain relief. Detection of the epidural space is critical, but it is difficult to predict the depth of the epidural space. Published results are inconsistent. We retrospectively investigated the differences in the depth of the epidural space depending on the puncture site, approach type and physical findings of patients. METHODS: All surgical patients from 1994 to 2005 were included in the study and 4964 cases were analysed. The recorded distance from the needle tip to the skin surface was defined as the depth of the epidural. The differences in the depth were compared according to the puncture site, level and approach. We then searched for the factors determining the depth using multivariate regression analysis. RESULTS: The depth in upper thoracic sites (T1/2-T9/10) was 5.0 (1.0) cm [mean (SD), n= 465] via the midline approach and 5.2 (1.0) cm (n= 1226) via the paramedian approach. For lumbar sites, the depth was 4.1 (0.9) cm (n= 1835) via the midline approach and 4.6 (1.0) cm (n= 298) via the paramedian approach. In the multivariate regression analysis, patient age, body weight and more cephalad puncture were significantly and positively correlated with the depth at thoracic sites. CONCLUSIONS: The epidural space was deeper at upper thoracic sites than lower thoracic or lumbar sites. The depth with the paramedian approach was greater than with the midline approach at both sites. Patient age and weight were positive factors for depth. PMID- 17425620 TI - Application of multi-locus analytical methods to identify interacting loci in case-control studies. AB - To identify interacting loci in genetic epidemiological studies the application of multi-locus methods of analysis is warranted. Several more advanced classification methods have been developed in the past years, including multiple logistic regression, sum statistics, logic regression, and the multifactor dimensionality reduction method. The objective of our study was to apply these four multi-locus methods to simulated case-control datasets that included a variety of underlying statistical two-locus interaction models, in order to compare the methods and evaluate their strengths and weaknesses. The results showed that the ability to identify the interacting loci was generally good for the sum statistic method, the logic regression and MDR. The performance of the logistic regression was more dependent on the underlying model and multiple comparison adjustment procedure. However, identification of the interacting loci in a model with two two-locus interactions of common disease alleles with relatively small effects was impaired in all methods. Several practical and methodological issues that can be considered in the application of these methods, and that may warrant further research, are identified and discussed. PMID- 17425621 TI - Immunity, homing and efficacy of allogeneic adoptive immunotherapy for posttransplant lymphoproliferative disorders. AB - Adoptive immunotherapy using autologous Epstein-Barr virus (EBV)-specific cytotoxic T-lymphocytes (auto-CTL) can regress posttransplant lymphoproliferative disorders (PTLD). Widespread applicability of auto-CTL remains constrained. Generation is time-consuming, and auto-CTL cannot be established in patients treated with the B-cell depleting antibody rituximab. By contrast, pregenerated allogeneic CTL (allo-CTL) offers immediate accessibility. Allo-CTL has previously shown efficacy in "early" polyclonal- PTLD. We treated three patients with aggressive, advanced monoclonal-PTLD following solid-organ transplantation. All were refractory to at least three prior therapies. Despite HLA disparity, there was negligible toxicity, with early in vivo antiviral efficacy and reconstitution of EBV peptide-specific immunity. Two patients attained complete remission (CR). One remains in CR 17 months following therapy, coincident with persistence of donor-derived tumor targeted EBV-specific CTL; the other died of non-PTLD related pathology. In the third patient, autopsy demonstrated homing of allo-CTL at the tumor site. Larger prospective studies of EBV-specific allo-CTL in PTLD are warranted. PMID- 17425622 TI - Tubular basement membrane immune deposits in association with BK polyomavirus nephropathy. AB - Tubular basement membrane immune deposits (TBMID) are rare in renal allografts and usually have been found in association with immune complex mediated glomerular injury. We report an association between TBMID and BK polyomavirus nephropathy (BKN). We reviewed clinical data and results of allograft biopsies of 30 patients with BKN (16 with and 14 without TBMID). TBMID were detected by immunofluorescence or electron microscopy. Initial and follow-up biopsies were assessed for degree of interstitial inflammation and fibrosis and severity of viral infection, and were correlated with patients' clinical data. Biopsies initially diagnostic for BKN with TBMID, compared to BKN biopsies without deposits, demonstrated more severe interstitial inflammation and fibrosis, and greater numbers of virally infected cells. Similar findings were present in follow-up biopsies. Utilizing three different antibodies directed against viral epitopes, viral antigens could not be detected within TBMID. Thirty percent of patients with TBMID and 70% without deposits had follow-up biopsies, in which virus could not be detected immunohistochemically. Treatment for all included decreasing immunosuppression, cidofovir and/or leflunomide. Clinical data correlated well with histological findings. We conclude that a significant proportion of patients with BKN show TBMID on kidney biopsy. The prognostic significance of this finding remains to be elucidated. PMID- 17425623 TI - Financial implications of pancreas transplant complications: a business case for quality improvement. AB - We quantified the financial implications of surgical complications following pancreas transplantation. We reviewed medical and financial records of 49 pancreas transplant recipients at the University of Michigan Health System (UMHS) between 1/6/2002 and 11/22/2004. The association of donor, transplant recipient and financial variables was assessed. The median costs to UMHS of procedures and follow-up were $92,917 for recipients without surgical complications versus $108,431 when a surgical complication occurred, a difference of $15,514 (p = 0.03). Median reimbursement by the payer was $17,363 higher in patients with a surgical complication (p = 0.001). Similar trends (higher insurer costs) were noted when stratifying by payer (public and private) and specific procedure (SPK and PAK). All parties (patient, physician, payer and medical center) should benefit from quality improvement, with payers having a financial interest in pancreas transplant surgical quality initiatives. PMID- 17425624 TI - Severe local vancomycin induced skin necrosis. PMID- 17425625 TI - Population pharmacokinetics and bioavailability of tacrolimus in kidney transplant patients. AB - AIMS: The use of tacrolimus is complicated by its narrow therapeutic index and wide intra- and interpatient variability. Tacrolimus population pharmacokinetics, including bioavailability, were investigated in an adult kidney transplant cohort to identify patient characteristics that influence pharmacokinetics. METHODS: The database (drug monitoring data) included 83 adult kidney transplant recipients and analysis was performed by a population approach with NONMEM. Data were collected during the first months after transplantation. Patients were administered oral or intravenous tacrolimus as part of a triple immunosuppressive regimen that also included mycophenolate mofetil and corticosteroids. Subsequent doses were adjusted on the basis of clinical evidence of efficacy and toxicity as in routine therapeutic drug monitoring. RESULTS: A one compartment open model with linear absorption and elimination adequately described the data. The typical value of minimal clearance was 1.8 +/- 0.2 l h(-1). Clearance increased with time post transplantation to reach 50% of maximal value after 3.8 +/- 0.5 days, with a maximal value of 5.6 l h(-1). Moreover clearance increased by approximately 1.6 fold (range 0.5-1.6) if the dose of prednisone was >25 mg. The typical value for volume of distribution, V, (98 +/- 13 l kg(-1)) was similar to reported values in kidney transplant patients. The oral bioavailability of tacrolimus was poor and ranged from 11.2 to 19.1%. No covariates significantly influenced V or F. CONCLUSIONS: The number of days postoperation and corticosteroid dose were significant covariates influencing tacrolimus clearance. PMID- 17425626 TI - Investigation on pharmacokinetics of mycophenolic acid in Chinese adult renal transplant patients. PMID- 17425627 TI - Effect of renal impairment on the pharmacokinetics of exenatide. AB - AIMS: To evaluate the pharmacokinetics (PK), safety and tolerability of a single exenatide dose in patients with renal impairment (RI). METHODS: Exenatide (5 or 10 microg) was injected subcutaneously in 31 subjects (one with Type 2 diabetes) stratified by renal function [Cockcroft-Gault creatinine clearance (CrCL), number of subjects]: normal (>80 ml min(-1), n = 8), mild RI (51-80 ml min(-1), n = 8), moderate RI (31-50 ml min(-1), n = 7) or end-stage renal disease (ESRD) requiring haemodialysis (n = 8). PK data were combined with four previous single-dose studies in patients with Type 2 diabetes to explore the relationship of exenatide clearance (CLp/F) and CrCL. RESULTS: Mean half-life for healthy, mild RI, moderate RI and ESRD groups were 1.5, 2.1, 3.2 and 6.0 h, respectively. After combining data from multiple studies, least squares geometric means for CLp/F in subjects with normal renal function, mild RI, moderate RI and ESRD were 8.14, 5.19, 7.11 and 1.3 l h(-1), respectively. Exenatide was generally well tolerated in the mild and moderate RI groups, but not in subjects with ESRD due to nausea and vomiting. Simulations of exenatide plasma concentrations also suggest patients with ESRD should have a propensity for poor tolerability at the lowest available therapeutic dosage (5 microg q.d.). CONCLUSIONS: Since tolerability and PK changes were considered clinically acceptable in patients with mild to moderate RI, it would be appropriate to administer exenatide to these patients without dosage adjustment. However, poor tolerability and significant changes in PK make the currently available therapeutic doses (5 and 10 microg) unsuitable in severe RI or ESRD. PMID- 17425628 TI - Effect of timing of dosing in relation to food intake on the pharmacokinetics of esomeprazole. AB - AIM: To investigate the pharmacokinetics of esomeprazole before a high-fat meal vs. fasting. METHODS: This open-label, randomized, crossover study consisted of two 5-day dosing periods of esomeprazole 40 mg per day. On days 1 and 5, subjects received esomeprazole 15 min before a high-fat meal (fed) or 4 h before a non high-fat meal (fasting). RESULTS: On days 1 and 5, ratio of fed to fasting area under the plasma concentration-time curve [0.56, 90% confidence interval (CI) 0.50, 0.64, and 0.78, 90% CI 0.74, 0.82, respectively] and peak plasma concentration (0.34, 90% CI 0.28, 0.41, and 0.47, 90% CI 0.41, 0.52, respectively) were outside of the limits of bioequivalence. CONCLUSIONS: Esomeprazole bioavailability was reduced when taken within 15 min before eating a high-fat meal vs. that while fasting. PMID- 17425629 TI - Randomized exposure-controlled trials; impact of randomization and analysis strategies. AB - AIMS: In the literature, five potential benefits of randomizing clinical trials on concentration levels, rather than dose, have been proposed: (i) statistical study power will increase; (ii) study power will be less sensitive to high variability in the pharmacokinetics (PK); (iii) the power of establishing an exposure-response relationship will be robust to correlations between PK and pharmacodynamics (PD); (iv) estimates of the exposure-response relationship are likely to be less biased; and (v) studies will provide a better control of exposure in situations with toxicity issues. The main aim of this study was to investigate if these five statements are valid when the trial results are evaluated using a model-based analysis. METHODS: Quantitative relationships between drug dose, concentration, biomarker and clinical end-point were defined using pharmacometric models. Three randomization schemes for exposure-controlled trials, dose-controlled (RDCT), concentration-controlled (RCCT) and biomarker controlled (RBCT), were simulated and analysed according to the models. RESULTS: (i) The RCCT and RBCT had lower statistical power than RDCT in a model-based analysis; (ii) with a model-based analysis the power for an RDCT increased with increasing PK variability; (iii) the statistical power in a model-based analysis was robust to correlations between CL and EC(50) or E(max); (iv) under all conditions the bias was negligible (<3%); and (v) for studies with equal power RCCT could produce either more or fewer adverse events compared with an RDCT. CONCLUSION: Alternative randomization schemes may not have the proposed advantages if a model-based analysis is employed. PMID- 17425630 TI - Flavonoids inhibit the platelet TxA(2) signalling pathway and antagonize TxA(2) receptors (TP) in platelets and smooth muscle cells. AB - AIMS: Flavonoids may affect platelet function by several mechanisms, including antagonism of TxA(2) receptors (TP). These TP are present in many tissues and modulate different signalling cascades. We explored whether flavonoids affect platelet TP signalling, and if they bind to TP expressed in other cell types. METHODS: Platelets were treated with flavonoids, or other selected inhibitors, and then stimulated with U46619. Similar assays were performed in aspirinized platelets activated with thrombin. Effects on calcium release were analysed by fluorometry and changes in whole protein tyrosine phosphorylation and activation of ERK 1/2 by Western blot analysis. The binding of flavonoids to TP in platelets, human myometrium and TPalpha- and TPbeta-transfected HEK 293T cells was explored using binding assays and the TP antagonist (3)H-SQ29548. RESULTS: Apigenin, genistein, luteolin and quercetin impaired U46619-induced calcium mobilization in a concentration-dependent manner (IC(50) 10-30 microm). These flavonoids caused a significant impairment of U46619-induced platelet tyrosine phosphorylation and of ERK 1/2 activation. By contrast, in aspirin-treated platelets all these flavonoids, except quercetin, displayed minor effects on thrombin-induced calcium mobilization, ERK 1/2 and total tyrosine phosphorylation. Finally, apigenin, genistein and luteolin inhibited by >50% (3)H SQ29548 binding to different cell types. CONCLUSIONS: These data further suggest that flavonoids may inhibit platelet function by binding to TP and by subsequent abrogation of downstream signalling. Binding of these compounds to TP occurs in human myometrium and in TP-transfected HEK 293T cells and suggests that antagonism of TP might mediate the effects of flavonoids in different tissues. PMID- 17425631 TI - Consumption of herbal remedies and dietary supplements amongst patients hospitalized in medical wards. AB - AIMS: Herbal remedies may have adverse effects and potentially serious interactions with some commonly prescribed conventional medications. Little is known about consumption of herbal remedies and dietary supplements by hospitalized patients. The aim was to evaluate the rate of consumption and characterize the patients hospitalized in internal medicine departments who consume herbal remedies and dietary supplements. Also, to assess the medical teams' awareness and assess the percentage of patients with possible drug-herb interactions. METHODS: Patients hospitalized in the medical wards of two hospitals in Israel were interviewed about their use of herbal remedies or dietary supplements. The medical records were searched for evidence that the medical team had knowledge of the use of herbal remedies or dietary supplements. RESULTS: Two hundred and ninety-nine hospitalized medical patients were interviewed. Of the participants, 26.8% were herbal or dietary supplement consumers (HC). On multivariate analysis the only variates associated with herbal or dietary supplement consumption were the hospital [odds ratio (OR) 2.97, 95% confidence interval (CI) 1.29, 6.52], income (OR 0.39, 95% CI 0.15, 1.05), smoking habits (OR 0.17, 95% CI 0.05, 0.55) and benign prostatic hypertrophy (OR 4.64, 95% CI 1.3, 16.5). Ninety-four percent of the patients had not been asked specifically of herbal consumption by the medical team. Only 23% of the hospital's medical files of the HC patients had any record of the use of herbal or dietary supplements. Seven possible drug-herbal interactions were encountered (7.1%). The most serious was an interaction between camomile tea and ciclosporin. CONCLUSIONS: Herbal remedy consumption is common amongst patients hospitalized in internal medicine wards and is often overlooked by the medical team. Patients and doctors should be more aware of the possible adverse effects and of the potential of herb-drug interactions. PMID- 17425632 TI - Joint analysis of longitudinal data with informative right censoring. AB - Longitudinal data arise when subjects are followed over a period of time. A commonly encountered complication in the analysis of such data is the variable length of follow-up due to right censorship. This can be further exacerbated by the possible dependency between the censoring time and the longitudinal measurements. This article proposes a combination of a semiparametric transformation model for the censoring time and a linear mixed effects model for the longitudinal measurements. The dependency is handled via latent variables which are naturally incorporated. We show that the likelihood function has an explicit form and develops a two-stage estimation procedure to avoid direct maximization over a high-dimensional parameter space. The resulting estimators are shown to be consistent and asymptotically normal, with a closed form for the variance-covariance matrix that can be used to obtain a plug-in estimator. Finite sample performance of the proposed approach is assessed through extensive simulations. The method is applied to renal disease data. PMID- 17425633 TI - Cox regression methods for two-stage randomization designs. AB - Two-stage randomization designs (TSRD) are becoming increasingly common in oncology and AIDS clinical trials as they make more efficient use of study participants to examine therapeutic regimens. In these designs patients are initially randomized to an induction treatment, followed by randomization to a maintenance treatment conditional on their induction response and consent to further study treatment. Broader acceptance of TSRDs in drug development may hinge on the ability to make appropriate intent-to-treat type inference within this design framework as to whether an experimental induction regimen is better than a standard induction regimen when maintenance treatment is fixed. Recently Lunceford, Davidian, and Tsiatis (2002, Biometrics 58, 48-57) introduced an inverse probability weighting based analytical framework for estimating survival distributions and mean restricted survival times, as well as for comparing treatment policies at landmarks in the TSRD setting. In practice Cox regression is widely used and in this article we extend the analytical framework of Lunceford et al. (2002) to derive a consistent estimator for the log hazard in the Cox model and a robust score test to compare treatment policies. Large sample properties of these methods are derived, illustrated via a simulation study, and applied to a TSRD clinical trial. PMID- 17425634 TI - Approximations for expected generation number. AB - A deterministic formula is commonly used to approximate the expected generation number of a population of growing cells. However, this can give misleading results because it does not allow for natural variation in the times that individual cells take to reproduce. Here we present more accurate approximations for both symmetric and asymmetric cell division. Based on the first two moments of the generation time distribution, these approximations are also robust. We illustrate the improved approximations using data that arise from monitoring individual yeast cells under a microscope and also demonstrate how the approximations can be used when such detailed data are not available. PMID- 17425635 TI - Applications and extensions of Chao's moment estimator for the size of a closed population. AB - This article revisits Chao's (1989, Biometrics45, 427-438) lower bound estimator for the size of a closed population in a mark-recapture experiment where the capture probabilities vary between animals (model M(h)). First, an extension of the lower bound to models featuring a time effect and heterogeneity in capture probabilities (M(th)) is proposed. The biases of these lower bounds are shown to be a function of the heterogeneity parameter for several loglinear models for M(th). Small-sample bias reduction techniques for Chao's lower bound estimator are also derived. The application of the loglinear model underlying Chao's estimator when heterogeneity has been detected in the primary periods of a robust design is then investigated. A test for the null hypothesis that Chao's loglinear model provides unbiased abundance estimators is provided. The strategy of systematically using Chao's loglinear model in the primary periods of a robust design where heterogeneity has been detected is investigated in a Monte Carlo experiment. Its impact on the estimation of the population sizes and of the survival rates is evaluated in a Monte Carlo experiment. PMID- 17425636 TI - Proximity model for expression quantitative trait loci (eQTL) detection. AB - Expression quantitative trait loci (eQTL) are loci or markers on the genomes that are associated with gene expression. It is well known to biologists that some (cis) genetic influences on expression occur over short distances on the genome while some (trans) influences can operate remotely. We use a log-linear model to place structure on the prior probability for genetic control of a transcript by a marker locus so that the loci that are closest to a transcript are given a higher prior probability of controlling that transcript to reflect the important role that genomic proximity can play in the regulation of expression. This proximity model is an extension of the mixture over marker (MOM) model for the simultaneous detection of cis and trans eQTL of Kendziorski (Kendziorski et al., 2006, Biometrics62(1), 19-27). The genomic locations of the transcripts are used to improve the accuracy of the posterior distribution for the location of the eQTL. We compare the MOM method to our extension with both simulated data and data sets of recombinant inbred mouse lines. We also discuss an extension of the MOM method to model multiple eQTLs, and find that many transcripts are likely associated with more than one eQTL. PMID- 17425637 TI - Modeling Maternal-Infant HIV Transmission in the Presence of Breastfeeding with an Imperfect Test. AB - An important public health question is to determine the probabilities of perinatal HIV transmission and when it occurs, whether antepartum, intrapartum, or postpartum through breastfeeding. However, this is a difficult problem because the presence of HIV infection in an infant can only be ascertained through viral assays in the postpartum period. We propose a model that simultaneously estimates the risks of antepartum, intrapartum, and postpartum transmissions together with the sensitivity of the screening tests for HIV infection. The model allows estimating of infectivity through breast milk during postpartum periods. The methods are illustrated on a South African randomized clinical trial of extended AZT versus a short course of nevirapine in infants whose mothers had no access to antenatal antiretroviral therapy. PMID- 17425638 TI - Time-varying coefficient models for the analysis of air pollution and health outcome data. AB - In this article a time-varying coefficient model is developed to examine the relationship between adverse health and short-term (acute) exposure to air pollution. This model allows the relative risk to evolve over time, which may be due to an interaction with temperature, or from a change in the composition of pollutants, such as particulate matter, over time. The model produces a smooth estimate of these time-varying effects, which are not constrained to follow a fixed parametric form set by the investigator. Instead, the shape is estimated from the data using penalized natural cubic splines. Poisson regression models, using both quasi-likelihood and Bayesian techniques, are developed, with estimation performed using an iteratively re-weighted least squares procedure and Markov chain Monte Carlo simulation, respectively. The efficacy of the methods to estimate different types of time-varying effects are assessed via a simulation study, and the models are then applied to data from four cities that were part of the National Morbidity, Mortality, and Air Pollution Study. PMID- 17425639 TI - Spatial multistate transitional models for longitudinal event data. AB - Follow-up medical studies often collect longitudinal data on patients. Multistate transitional models are useful for analysis in such studies where at any point in time, individuals may be said to occupy one of a discrete set of states and interest centers on the transition process between states. For example, states may refer to the number of recurrences of an event, or the stage of a disease. We develop a hierarchical modeling framework for the analysis of such longitudinal data when the processes corresponding to different subjects may be correlated spatially over a region. Continuous-time Markov chains incorporating spatially correlated random effects are introduced. Here, joint modeling of both spatial dependence as well as dependence between different transition rates is required and a multivariate spatial approach is employed. A proportional intensities frailty model is developed where baseline intensity functions are modeled using parametric Weibull forms, piecewise-exponential formulations, and flexible representations based on cubic B-splines. The methodology is developed within the context of a study examining invasive cardiac procedures in Quebec. We consider patients admitted for acute coronary syndrome throughout the 139 local health units of the province and examine readmission and mortality rates over a 4-year period. PMID- 17425640 TI - Determining the number of clusters using the weighted gap statistic. AB - Estimating the number of clusters in a data set is a crucial step in cluster analysis. In this article, motivated by the gap method (Tibshirani, Walther, and Hastie, 2001, Journal of the Royal Statistical Society B63, 411-423), we propose the weighted gap and the difference of difference-weighted (DD-weighted) gap methods for estimating the number of clusters in data using the weighted within clusters sum of errors: a measure of the within-clusters homogeneity. In addition, we propose a "multilayer" clustering approach, which is shown to be more accurate than the original gap method, particularly in detecting the nested cluster structure of the data. The methods are applicable when the input data contain continuous measurements and can be used with any clustering method. Simulation studies and real data are investigated and compared among these proposed methods as well as with the original gap method. PMID- 17425641 TI - Confidence intervals for uncommon but dramatic responses to treatment. AB - A small literature discusses locally most powerful rank tests when only a fraction of treated subjects respond to treatment. The ranks used in these tests are very different from conventional ranks, being relatively flat for low responses and then rising steeply, and the associated tests are much more powerful than conventional rank tests when, indeed, only a small fraction of treated subjects exhibit dramatic responses. Because the tests are derived from considerations of local power, they do not yield a plausible family of models for effect, and therefore they do not yield confidence intervals for the magnitude of effect formed by inverting the tests. There is a similarity between these tests and another family of tests, originally motivated by different considerations involving peak performance in small subsets. Exploiting this similarity, a method for obtaining confidence statements is proposed. In the case of observational studies, sensitivity to unobserved bias from nonrandom assignment of treatments is also examined. Two examples are used as illustrations: (i) a study of smoking during pregnancy and its effects on birth weight, in which smokers are matched to six controls, and (ii) a matched pair study of damage to DNA among workers at aluminum production plants. PMID- 17425642 TI - Type I and Type II error under random-effects misspecification in generalized linear mixed models. AB - Generalized linear mixed models (GLMMs) have become a frequently used tool for the analysis of non-Gaussian longitudinal data. Estimation is based on maximum likelihood theory, which assumes that the underlying probability model is correctly specified. Recent research is showing that the results obtained from these models are not always robust against departures from the assumptions on which these models are based. In the present work we have used simulations with a logistic random-intercept model to study the impact of misspecifying the random effects distribution on the type I and II errors of the tests for the mean structure in GLMMs. We found that the misspecification can either increase or decrease the power of the tests, depending on the shape of the underlying random effects distribution, and it can considerably inflate the type I error rate. Additionally, we have found a theoretical result which states that whenever a subset of fixed-effects parameters, not included in the random-effects structure equals zero, the corresponding maximum likelihood estimator will consistently estimate zero. This implies that under certain conditions a significant effect could be considered as a reliable result, even if the random-effects distribution is misspecified. PMID- 17425643 TI - Cutaneous cryptococcosis with alemtuzumab in a patient treated for chronic lymphocytic leukaemia. PMID- 17425644 TI - Irregularly shaped leucoplakic lesion of the anus. Diagnosis: anal intraepithelial neoplasia (AIN). PMID- 17425645 TI - Coincidental presentation of vitiligo and psoriasis in a patient with polyglandular autoimmune syndrome. PMID- 17425647 TI - Kumkum-induced dermatitis: an analysis of 46 cases. AB - Kumkum is a commonly used cosmetic in India, traditionally based on turmeric powder, and its ingredients can cause various types of dermatosis. We undertook a study to characterize kumkum-induced dermatitis clinically and to identify by patch testing the allergens involved. In total, 46 patients with kumkum-induced dermatitis (mean age of these cases 46.5 years; female : male ratio 1.8 : 1) were investigated. The forehead was the most commonly involved site (31/46), followed by the glabellar area (16/46), hair parting (6/46), abdomen (5/46), and neck (3/46). Pigmented contact dermatitis was seen in 35 patients (76.1%), and allergic contact dermatitis in 11 patients (23.9%). In total, 25 patients were patch tested. The commonest positive allergic reaction was to thimerosal (18/25, 72%), followed by gallate mix (12/25, 48%). Para-phenylenediamine, Kathon CG, benzotriazol, tert-butyl hydroquinone and parabens gave a reaction for one patient each (1/25, 4%). Patch testing with kumkum was positive in 7 of 13 patients tested. PMID- 17425648 TI - Plantar pressures are higher under callused regions of the foot in older people. AB - BACKGROUND: Mechanically induced hyperkeratotic lesions (corns and calluses) are among the most common foot problems in older people. However, their aetiology is not well understood. AIM: To compare the magnitude of pressures generated under the foot when walking in older people with and without plantar calluses. METHODS: Peak plantar pressure measurements were obtained from 292 participants (99 men and 193 women) aged 62-96 years (mean +/- SD 77.6 +/- 6.9) recruited from a retirement village and a university health sciences clinic. Comparisons were then made between callused and noncallused regions of the foot. RESULTS: In total, 151 participants (52%) had at least one plantar callus. Those with plantar calluses were more likely to be female, have moderate to severe hallux valgus, and at least one lesser toe deformity. Regional peak plantar pressures were significantly higher in people with calluses under the second metatarsophalangeal joint (2.34 +/- 0.46 vs. 2.12 +/- 0.51 kg/cm(2), P = 0.001), the third to fifth metatarsophalangeal joints (1.71 +/- 0.46 vs. 1.50 +/- 0.51 kg/cm(2), P = 0.009) and the hallux (1.40 +/- 0.34 vs. 1.23 +/- 0.47 kg/cm(2), P = 0.007) compared with people without calluses under these sites. CONCLUSION: Plantar pressures are significantly higher under callused regions of the foot in older people. Raised pressure may play a role in the development of plantar calluses by accelerating the turnover rate of keratinocytes in the epidermis. Future studies should focus on evaluating the efficacy of pressure-relieving interventions in the prevention and treatment of keratotic disorders in older people. PMID- 17425649 TI - Which aeroallergens are associated with eczema severity? AB - We investigated if a correlation exists between aeroallergen sensitization and the severity of eczema. Data on aeroallergen response to skin-prick testing (SPT) and disease severity of children with eczema (n = 119) were evaluated. Atopy, as defined by at least one positive response to aeroallergen skin prick testing, was found in > 90% of eczema patients. House dust mite was the most commonly sensitized aeroallergen, followed by cat fur. Dermatophagoides pteronyssinus and Dermatophagoides farinae sensitization were associated with eczema severity (present in 67% of the mild and 97% of the severe group; P = 0.001). However, there was no association between eczema severity and higher strengths of SPT response (defined as SPT > 1+ to dust mites or dust). Atopy to moulds, Bermuda grass, cockroach, cat and dog was less prevalent and was not associated with eczema severity. It is sensible to advise parents on specific avoidance strategies only in severely affected children who have a definitive history of eczema exacerbation by specific aeroallergens and who are not responsive to conventional treatment. PMID- 17425650 TI - Methicillin-resistant Staphylococcus aureus pyomyositis presenting with cellulitis-like symptoms. PMID- 17425651 TI - Smoking and a complement gene polymorphism interact in promoting cardiovascular disease morbidity and mortality. AB - We have demonstrated previously that carriers of a genotype called C4B*Q0 (silent allele of the C4B gene) have a substantially increased risk to suffer from myocardial infarction or stroke, and are selected out from the healthy elderly population. Because smoking carries a major risk for cardiovascular disease (CVD), it seemed worthwhile to study if these two factors interact. Study 1 involved 74 patients with angina pectoris (AP), 85 patients with recent acute myocardial infarction (AMI) and 112 survivors of a previous AMI and 382 controls from Iceland. Study 2 involved 233 patients with severe CVD and 274 controls from Hungary. Smoking habits were registered for each subject. The number of C4A and C4B genes was determined by phenotyping or genotyping. Compared to controls, C4B*Q0 carrier frequency was significantly higher at diagnosis in Icelandic smokers with AP (P = 0.005) and AMI (P = 0.0003) and Hungarian smokers with severe coronary artery disease (P = 0.023), while no such difference was observed in non-smoking subjects. Age-associated decrease in C4B*Q0 observed previously in two remote Caucasian populations was found, in the present study, to be associated strongly with smoking, and to already occur in smokers after age 50 years both in Iceland and Hungary. Our findings indicate that the C4B*Q0 genotype can be considered as a major covariate of smoking in precipitating the risk for AMI and associated deaths. PMID- 17425652 TI - Low mannose-binding lectin complement activation function is associated with predisposition to Legionnaires' disease. AB - Innate immune system deficiency may predispose to severe infections such as Legionnaires' disease. We have investigated the role of mannose-binding lectin (MBL) deficiency in the Melbourne Aquarium Legionnaires' disease outbreak. Serum samples from patients and controls that were exposed but shown to be uninfected from the Melbourne Aquarium Legionnaires' disease outbreak were tested for MBL function (C4 deposition) and level (mannan-binding). MBL function was lower in Legionnaires' disease cases than in age- and sex-matched uninfected, exposed controls. The frequency of MBL deficiency with C4 deposition < 0.2 U/microl was significantly higher in Legionnaires' disease cases than in controls. This also applied to Legionnaires' disease cases requiring hospital care. There was no difference in MBL mannan-binding levels between Legionnaires' disease patients and controls. There was no significant interval change in MBL function or level after a mean of 46 days. MBL complement activation functional deficiency appears to predispose to Legionnaires' disease. PMID- 17425653 TI - Aberrant activation profile of cytokines and mitogen-activated protein kinases in type 2 diabetic patients with nephropathy. AB - Cytokine-induced inflammation is involved in the pathogenesis of type 2 diabetes mellitus (DM). We investigated plasma concentrations and ex vivo production of cytokines and chemokines, and intracellular signalling molecules, mitogen activated protein kinases (MAPK) in T helper (Th) cells and monocytes in 94 type 2 diabetic patients with or without nephropathy and 20 healthy controls. Plasma concentrations of inflammatory cytokines tumour necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-18 and chemokine CCL2 in patients with diabetic nephropathy (DN) were significantly higher than control subjects, while IL-10, CXCL8, CXCL9, CXCL10 and adiponectin concentrations of DN were significantly higher than patients without diabetic nephropathy (NDN) and control subjects (all P < 0.05). Plasma concentrations of TNF-alpha, IL-6, IL-10, IL-18, CCL2, CXCL8, CXCL9, CXCL10 and adiponectin exhibited significant positive correlation with urine albumin : creatinine ratio in DN patients. The percentage increases of ex vivo production of IL-6, CXCL8, CXCL10, CCL2 and CCL5 upon TNF-alpha activation were significantly higher in both NDN and DN patients than controls (all P < 0.05). The percentage increases in IL-18-induced phosphorylation of extracellular signal-regulated kinase (ERK) in Th cells of NDN and DN were significantly higher than controls (P < 0.05), while the percentage increase in TNF-alpha-induced phosphorylation of p38 MAPK in monocytes and IL-18-induced phosphorylation of p38 MAPK in Th cells and monocytes were significantly higher in NDN patients than controls. These results confirmed that the aberrant production of inflammatory cytokines and chemokines and differential activation of MAPK in different leucocytes are the underlying immunopathological mechanisms of type 2 DM patients with DN. PMID- 17425654 TI - Skewed T cell receptor repertoire of Vdelta1(+) gammadelta T lymphocytes after human allogeneic haematopoietic stem cell transplantation and the potential role for Epstein-Barr virus-infected B cells in clonal restriction. AB - The proliferation of Vdelta1(+) gammadelta T lymphocytes has been described in various infections including human immunodeficiency virus (HIV), cytomegalovirus (CMV) and malaria. However, the antigen specificity and functions of the human Vdelta1(+) T cells remain obscure. We sought to explore the biological role for this T cell subset by investigating the reconstitution of T cell receptor (TCR) repertoires of Vdelta1(+) gammadelta T lymphocytes after human allogeneic haematopoietic stem cell transplantation (HSCT). We observed skewed TCR repertoires of the Vdelta1(+) T cells in 27 of 44 post-transplant patients. Only one patient developed EBV-associated post-transplant lymphoproliferative disorder in the present patient cohort. The -WGI- amino acid motif was observed in CDR3 of clonally expanded Vdelta1(+) T cells in half the patients. A skew was also detected in certain healthy donors, and the Vdelta1(+) T cell clone derived from the donor mature T cell pool persisted in the recipient's blood even 10 years after transplant. This T cell clone expanded in vitro against stimulation with autologous EBV-lymphoblastoid cell lines (LCL), and the Vdelta1(+) T cell line expanded in vitro from the same patient showed cytotoxicity against autologous EBV-LCL. EBV-infected cells could also induce in vitro oligoclonal expansions of autologous Vdelta1(+) T cells from healthy EBV-seropositive individuals. These results suggest that human Vdelta1(+) T cells have a TCR repertoire against EBV infected B cells and may play a role in protecting recipients of allogeneic HSCT from EBV-associated disease. PMID- 17425655 TI - BY55/CD160 cannot be considered a cytotoxic marker in cytomegalovirus-specific human CD8(+) T cells. AB - CD160/BY55 is a glucosyl-phosphatidylinositol (GPI)-anchored cell membrane receptor that is expressed primarily in natural killer (NK) cells. Its presence in CD8(+) T lymphocytes is considered to be a marker of cytotoxic activity, although there are few data in this regard. In the present work, we analysed the expression of CD160 in subpopulations of cytomegalovirus (CMV)-specific CD8(+) T cells. Subpopulations were defined by CD28 and CD57 expression and exhibited varying degrees of differentiation and cytotoxic potential, as evaluated by the expression of perforin, interferon (IFN)-gamma and interleukin (IL) 7Ralpha/CD127. We included subjects with different intensities of anti-viral immune response. Results showed that the terminally differentiated CD28(-) CD57(+) subset displaying the highest level of perforin expressed CD160 at a level similar to that of memory CD28(+) CD57(-)perforin(-) cells. A comparison of the expression of perforin in CD160(+) cells versus CD160(-) cells showed that expression was significantly higher in the absence of CD160. Interestingly, the CMV-specific CD8(+) T cell subset from a patient with ongoing CMV reactivation did not begin to express CD160 until day +92 of the follow-up period. Taken together, our data show that CD160 cannot be considered a cytotoxic marker in CMV specific CD8(+) T cells. PMID- 17425656 TI - Autoantibodies to plasminogen and tissue plasminogen activator in women with recurrent pregnancy loss. AB - Reduced fibrinolytic activity has been described in primary anti-phospholipid syndrome (PAPS), and may be responsible for thrombotic events. Antibodies to tissue type plasminogen activator (t-PA) or plasminogen (PLG) might contribute to the hypofibrinolytic state in autoimmune diseases, but the clinical significance of these antibodies is still unclear in recurrent pregnancy loss (RPL). The aim of this study is to evaluate the prevalence and clinical significance of anti-PLG and anti-t-PA antibodies in 87 patients with a history of RPL: 54 women with well defined PAPS (mean age 32.5 years; range 26-38) and 33 women with unexplained RPL (mean age 30 years; range 24-39). IgG anti-PLG antibodies were found in 20 and four patients from the group with RPL/PAPS and unexplained RPL, respectively; IgG anti-t-PA antibodies were found in 11 and two patients from the above two groups, respectively. IgG anti-PLG antibodies were associated with the high risk of RPL (OR 7.2, P = 0.004), especially with RPL/PAPS (OR 11.2, P < 0.001) evaluated by Fisher's exact test, while IgG anti-t-PA were associated with RPL/PAPS (OR 10.0, P = 0.01) but not with RPL (OR 6.8, P = 0.06). A significant inhibition of exogenous fibrinolysis was observed by IgG fractions from patients with anti-PLG or anti-t-PA antibodies on microplates and on the human umbilical vein endothelial cells, compared with those from healthy controls. The prevalence of IgG anti-PLG antibodies was high in RPL patients, especially in RPL/PAPS, while the prevalence of IgG anti-t-PA antibodies was high in RPL/PAPS but not in RPL, and some of them might inhibit fibrinolysis in patients. PMID- 17425657 TI - Increased bone formation to unstable nano rough titanium implants. AB - Early bone response to cylindrical smooth titanium implants (S(a)=0.1 microm) inserted into the rabbit tibia was compared in a stable and nonstable regime. Surface roughness parameters were calculated from measurements obtained with optical interferometry and atomic force microscopy. Contrary to our hypothesis, the nonstable implant showed higher bone to metal contact and increased bone area in the endosteal region compared with the stable implant after 4 weeks of healing. Bone area measurements in the cortical region revealed similar values. Primitive woven bone was found in close contact with both implants, but significantly more with the nonstable implant. Finding more bone-to-implant contact (BIC) need not necessarily indicate that unstable implants were more strongly integrated. Primitive bone stage development observed indicates less strong implant anchorage than could be expected from BIC percentage alone. Stable implant design used in this study is a reliable model to evaluate submicron and nanostructures in vivo, as implant stability was achieved in the absence of microirregularaties. PMID- 17425658 TI - Adrenocortical zonation factor 1 is a novel matricellular protein promoting integrin-mediated adhesion of adrenocortical and vascular smooth muscle cells. AB - Expression of a previously cloned secretory protein named adrenocortical zonation factor 1 (AZ-1, also called Tin-ag-RP or lipocalin 7) is tightly linked with the zonal differentiation of adrenocortical cells. It is also present in vascular smooth muscle (VSM), although its function has remained unknown. In this study, the location of AZ-1 was specified to the basal laminae along adrenocortical sinusoidal capillaries and surrounding VSM cells in the arterial system, consistent with the fact that AZ-1 was extractable under denaturing conditions as a 52 kDa polypeptide. Purified recombinant AZ-1 exhibited abilities to bind to fibronectins via the first type III repeat (anastellin) and to collagens with affinities in submicromolar ranges. AZ-1 immobilized on substratum or bound to collagens or anastellin promoted adhesion and spreading of adrenocortical cells. Although VSM cells spread on AZ-1 slowly, AZ-1 bound to anastellin facilitated the spreading. The adhesion activity of AZ-1 was mediated by a subset of integrins, including alpha(1)beta(1), alpha(2)beta(1), and alpha(5)beta(1), in a cell type-specific manner. Collectively with the putative role of AZ-1 in the adrenocortical zonation, we propose that AZ-1 potentially regulates functions of adrenocortical and VSM cells by modulating cell-matrix interactions. PMID- 17425659 TI - Primary administration of Lactobacillus johnsonii NCC533 in weaning period suppresses the elevation of proinflammatory cytokines and CD86 gene expressions in skin lesions in NC/Nga mice. AB - The administration of probiotic lactic acid bacteria (LAB) has been studied for its potential to prevent atopic dermatitis (AD). The objective of this study was to assess the inhibitory mechanism of a skin lesion by LAB using an experimental model that we previously demonstrated in NC/Nga mice. Lactobacillus johnsonii NCC533 (La1) was administered orally to the La1 group from 20 to 22 days after birth, while phosphate-buffered saline was given to the control group. After the induction of skin lesions in 6-week-old mice, the expression of genes supposedly involved in AD was evaluated. Gene expression of the proinflammatory cytokines [interleukin-8 (IL-8), IL-12 and IL-23] was significantly enhanced in the lesional skin of the control group by the induction of the lesion, whereas gene expression of those in the La1 group was not elevated. Interestingly, expression of the costimulatory molecule CD86 showed a pattern similar to the expression of the cytokines in the lesional skin. Moreover, the La1 group showed a significantly lower gene expression of CD86 in Peyer's patches and mesenteric lymph nodes than the control group. The suppression of proinflammatory cytokines and CD86 by primary administration of La1 may significantly contribute to the inhibitory effect on the skin lesion. PMID- 17425660 TI - Roseovarius sp. strain 217: aerobic taurine dissimilation via acetate kinase and acetate-CoA ligase. AB - The genome sequence of Roseovarius sp. strain 217 indicated that many pathway enzymes found in other organisms for the degradation of taurine are represented, but that a novel, apparently energy-dependent pathway is involved in the conversion of acetyl phosphate to acetyl CoA. Thus, an ABC transporter for taurine could be postulated, while inducible taurine: pyruvate aminotransferase, alanine dehydrogenase, sulfoacetaldehyde acetyltransferase and sulfite dehydrogenase could be assayed. Whereas phosphate acetyltransferase has been found in other organisms, none was indicated in the genome sequence and no activity was found in cell-free extracts. Instead, acetate kinase was active as was acetate-CoA ligase. PMID- 17425661 TI - Isolation and characterization of a carbofuran-degrading strain Novosphingobium sp. FND-3. AB - A gram-negative Novosphingobium sp. strain FND-3 capable of degrading carbofuran was isolated and characterized. The carbofuran-degrading ability of strain FND-3 was investigated under various culture conditions. Strain FND-3 showed a high average carbofuran-degrading rate of 28.6 mg L(-1) h(-1) in mineral salts medium with 100 mg L(-1) carbofuran. GC/MS analysis pointed out the presence of several unknown metabolites. One hydrolyzate was identified as 2-hydroxy-3-(3-methypropan 2-ol) phenol via hydrolysis of carbofuran phenol. The appearance of another metabolite with M(+) of 180 m/z indicated that the hydroxylation of carbofuran occurred at the aromatic ring. One novel degrading product with M(+) of 239 m/z was identified as 2-hydroxy-3-(3-methylpropan-2-ol) benzene-N-methylcarbamate via hydrolyzing at the ether bond of furanyl ring of carbofuran. Strain FND-3 was also able to degrade other N-methylcarbamate pesticides. PMID- 17425662 TI - Cloning and functional expression of the mitochondrial alternative oxidase of Aspergillus fumigatus and its induction by oxidative stress. AB - Aspergillus fumigatus possesses a branched mitochondrial electron transport chain, with both cyanide-sensitive and -insensitive oxygen-consumption activities. Mitochondrial reactive oxygen species mediate signaling for alternative oxidase (AOX) expression. A 1173 bp-long Afaox gene encoding a 40 kDa protein has been cloned and identified. Recombinant constructs containing the Afaox ORF were transformed into Escherichia coli and Saccharomyces cerevisiae for heterologous expression. In A. fumigatus, AOX activity and mRNA expression were both induced with menadione or paraquat, suggesting an important role of AOX under oxidative stress. Therefore, positive transformants showed a cyanide resistant and salicylhydroxamic acid-sensitive respiration, whereas in control cells the oxygen uptake was completely inhibited after KCN addition. PMID- 17425663 TI - ABO blood group, other risk factors and incidence of venous thromboembolism: the Longitudinal Investigation of Thromboembolism Etiology (LITE). AB - BACKGROUND: Numerous case-control studies have reported higher prevalence of non O blood type among venous thromboembolism (VTE) patients than controls, but potential mechanisms or effect modifiers for the association are not fully established. PATIENTS/METHODS: Using a nested case-control design combining the Atherosclerosis Risk in Communities and the Cardiovascular Health Study cohort, ABO blood type and other VTE risk factors were measured on pre-event blood samples of 492 participants who subsequently developed VTE and 1008 participants who remained free of VTE. RESULTS: A total of 64.4% of cases and 52.5% of controls had non-O blood type. Among controls, mean values of factor VIIIc (FVIIIc) and von Willebrand factor among the non-O blood type group were higher than among the O group. Compared with O blood type, the age-adjusted odds ratio (OR) of VTE for non-O blood type was 1.64 (95% CI, 1.32-2.05) and was similar for the two parent studies and race groups. Further adjustment for sex, race, body mass index, diabetes mellitus and FVIIIc reduced the OR: 1.31 (95% CI, 1.02 1.68). Factor V Leiden (FV Leiden) appeared to modify the non-O blood type association with VTE in a supra-additive fashion, with an age-, sex- and race adjusted OR of 6.77 (95% CI, 3.65-12.6) for having both risk factors. CONCLUSIONS: Non-O blood type was independently associated with risk of VTE, and added to the risk associated with FV Leiden. PMID- 17425664 TI - Leptin induces tissue factor expression in human peripheral blood mononuclear cells: a possible link between obesity and cardiovascular risk? AB - BACKGROUND: Obesity is a major modifiable risk factor for cardiovascular disease. Leptin, the hormone synthesized and released primarily by adipose tissue and found increased in obese individuals, has been implicated in the regulation of inflammation and arterial and venous thrombosis. OBJECTIVE: To investigate the role of tissue factor (TF), the pivotal agonist of the clotting cascade, as a link between obesity and cardiovascular disease. METHODS AND RESULTS: In 15 obese patients, plasma levels of leptin and TF as well as TF expression in resting and endotoxin-stimulated mononuclear leukocytes (MN) were increased when compared with healthy donors. In a selected sample of obese patients, loss of body weight led to decreased circulating leptin levels, accompanied by a reduction in plasma TF as well as in TF expression, both in resting and endotoxin-stimulated MN. In subsequent in vitro experiments, leptin was incubated with MN from healthy subjects. Leptin induced TF activity and antigen in a dose-dependent fashion, as assessed by clotting assay and ELISA, respectively. Increased migration of c Rel/p65 into the nucleus, as determined by EMSA, and development of TF mRNA in monocytes, as assessed by RT-PCR, were observed. Experiments with mitogen activated protein kinase (MAPK) inhibitors, indicated the involvement of p38 and ERK1/2 pathways. CONCLUSIONS: The presence of TF-expressing MN in blood from obese subjects and the in vitro induction of TF by pharmacologic concentrations of leptin in MN from healthy subjects suggest that TF expression by leptin stimulated monocytes may contribute to the cardiovascular risk associated with obesity. PMID- 17425665 TI - Changes in fibrinolytic activity after angiotensin II receptor blockade in therapy-resistant hypertensive patients. AB - BACKGROUND: In hypertensive patients, the activated renin-angiotensin system induces a prothrombotic state resulting from imbalance between coagulation and fibrinolysis. Although blood pressure cannot be regulated in therapy-resistant hypertensive patients, they may still be responsive to medication that attenuates the renin-angiotensin system. OBJECTIVE: our objective was to study possible attenuating properties of angiotensin II type 1 receptor blockers (AT1RBs) on the prothrombotic state in therapy-resistant hypertensive patients, focusing on parameters of fibrinolysis and coagulation. METHODS: Fourteen therapy-resistant hypertensive patients received AT1RB eprosartan infusion (45 and 150 microg kg( 1)) (study group), and 33 therapy-resistant hypertensive patients received saline (0.9%) infusion (control group) prior to renal angiography. Baseline values of parameters of coagulation and fibrinolysis were set at 1.00, and relative changes were calculated. RESULTS: Plasminogen activator inhibitor type 1 (PAI-1) antigen showed non-significant decreases in both the study group (arterial 1.00-0.45, venous 1.00-0.42) and control group (arterial 1.00-0.84, venous 1.00-0.88). PAI-1 activity significantly decreased in the study group (arterial 1.00-0.72, venous 1.00-0.71) and control group (arterial 1.00-0.83, venous 1.00-0.94). In the study group, tissue-type plasminogen activator (t-PA) antigen decreased significantly (arterial 1.00-0.62, venous 1.00-0.67), whereas t-PA activity significantly increased (arterial 1.00-6.15, venous 1.00-2.66). In the control group, t-PA antigen remained unchanged. No changes were observed in blood pressure during and after infusion of eprosartan. CONCLUSION: Therapy-resistant hypertensive patients show beneficial changes in fibrinolytic activity after infusion of a non-pressor dose of AT1RB. PMID- 17425666 TI - Tissue factor around dermal vessels has bound factor VII in the absence of injury. AB - BACKGROUND: 'Idling' or ongoing low-level activity of the tissue factor (TF) pathway is a postulated mechanism by which the coagulation process can become active without a lag period at sites of injury. OBJECTIVE: To determine whether TF around cutaneous vessels has bound factor VIIa in the absence of injury, and thus could participate in the idling process. METHODS: Immunostaining of mouse skin with antibodies against a 15-residue peptide from the sequence of mouse TF, and against the whole extracellular portion of TF. RESULTS: The whole TF antibody recognized TF in squamous epithelium and around vessels in the dermis. By contrast, the monospecific antibody only recognized TF in the squamous epithelium, but not around vessels. We also found that biotinylated, active site inhibited FVIIa (FVIIai) bound to tissue sections in the same areas in which TF was recognized by the monospecific antibody (squamous epithelium), but did not bind around vessels. Molecular modeling revealed that FVIIa and FX binding to TF masked a significant part of the surface of the target peptide. CONCLUSIONS: In the aggregate, these data are most consistent with the interpretation that TF in perivascular sites has bound FVIIa, even in the absence of any injury. The presence of endogenously bound FVIIa prevents the subsequent binding of the monospecific antibody or exogenous FVIIai to perivascular TF. PMID- 17425667 TI - A novel class of small repetitive DNA sequences in Enterococcus faecalis. AB - The structural organization of Enterococcus faecalis repeats (EFAR) is described, palindromic DNA sequences identified in the genome of the Enterococcus faecalis V583 strain by in silico analyses. EFAR are a novel type of miniature insertion sequences, which vary in size from 42 to 650 bp. Length heterogeneity results from the variable assembly of 16 different sequence types. Most elements measure 170 bp, and can fold into peculiar L-shaped structures resulting from the folding of two independent stem-loop structures (SLSs). Homologous chromosomal regions lacking or containing EFAR sequences were identified by PCR among 20 E. faecalis clinical isolates of different genotypes. Sequencing of a representative set of 'empty' sites revealed that 24-37 bp-long sequences, unrelated to each other but all able to fold into SLSs, functioned as targets for the integration of EFAR. In the process, most of the SLS had been deleted, but part of the targeted stems had been retained at EFAR termini. PMID- 17425668 TI - Analysis of Trichophyton rubrum gene expression in response to cytotoxic drugs. AB - Suppressive subtractive hybridization was used to isolate transcripts specifically upregulated during Trichophyton rubrum exposure to acriflavin, fluconazole, griseofulvin, terbinafine or undecanoic acid. Macro-array dot-blot and sequencing of 132 clones, which correspond to genes differentially expressed after exposition of T. rubrum to at least one of these cytotoxic drugs, revealed 39 unique genes. Of these, 32 have not been previously described in T. rubrum, representing an increase in the number of T. rubrum genes that have been identified. The upregulation of the novel genes encoding a retrotransposon element, a carboxylic ester hydrolase, a copper resistance-associated P-type ATPase, a DNA mismatch repair protein and a NIMA (never in mitosis A) interactive protein was confirmed by Northern blot. PMID- 17425669 TI - Central carbon metabolism of Saccharomyces cerevisiae in anaerobic, oxygen limited and fully aerobic steady-state conditions and following a shift to anaerobic conditions. AB - Saccharomyces cerevisiae CEN.PK113-1A was grown in glucose-limited chemostat culture with 0%, 0.5%, 1.0%, 2.8% or 20.9% O2 in the inlet gas (D=0.10 h(-1), pH 5, 30 degrees C) to determine the effects of oxygen on 17 metabolites and 69 genes related to central carbon metabolism. The concentrations of tricarboxylic acid cycle (TCA) metabolites and all glycolytic metabolites except 2 phosphoglycerate+3-phosphoglycerate and phosphoenolpyruvate were higher in anaerobic than in fully aerobic conditions. Provision of only 0.5-1% O2 reduced the concentrations of most metabolites, as compared with anaerobic conditions. Transcription of most genes analyzed was reduced in 0%, 0.5% or 1.0% O2 relative to cells grown in 2.8% or 20.9% O2. Ethanol production was observed with 2.8% or less O2. After steady-state analysis in defined oxygen concentrations, the conditions were switched from aerobic to anaerobic. Metabolite and transcript levels were monitored for up to 96 h after the transition, and this showed that more than 30 h was required for the cells to fully adapt to anaerobiosis. Levels of metabolites of upper glycolysis and the TCA cycle increased following the transition to anaerobic conditions, whereas those of metabolites of lower glycolysis generally decreased. Gene regulation was more complex, with some genes showing transient upregulation or downregulation during the adaptation to anaerobic conditions. PMID- 17425670 TI - The Saccharomyces cerevisiae Arf3 protein is involved in actin cable and cortical patch formation. AB - We show that Arf3p, a member of the ADP ribosylation family, is involved in the organization of actin cables and cortical patches in Saccharomyces cerevisiae. Profilin-deficient cells (pfy1Delta) have severe growth defects and lack actin cables. Overexpression of ARF3 restores actin cables and corrects growth defects in these cells. Cells deficient for the cortical patch proteins Las17p and Vrp1p have growth defects and a random cortical patch distribution. Overexpression of ARF3 in las17Delta and in vrp1Delta cells partially corrects growth defects and restores the polarized distribution of cortical patches. The N-terminal glycine, a myristoylation site in Arf3p, is necessary for its suppressor activity. arf3Delta cells show a random budding pattern. Overexpression of BNI1, GEA2 or SYP1, three genes involved in actin cytoskeleton formation, restores the normal axial budding pattern of arf3Delta cells. BUD6 is a polarity gene and GEA2 is involved in retrograde transport and the organization of the actin cytoskeleton. We have identified genetic interactions between ARF3 and BUD6, and between ARF3 and GEA2. Both double mutant strains have actin cytoskeleton defects. Our results support a role for ARF3 in cell polarity and the organization of the actin cytoskeleton. PMID- 17425671 TI - The use of killer sensitivity patterns for biotyping yeast strains: the state of the art, potentialities and limitations. AB - In recent years molecular techniques have been the most useful tools for the unequivocal identification of undetermined strains at the species level. In many instances, however, a further discrimination at the strain level (biotyping) is required, such as during epidemiological investigations, in which the distribution of pathogenic microorganisms is studied, and for patent protection purposes. Although molecular methods are routinely used also for yeast biotyping, several nonmolecular techniques have been proposed. One of these, the determination of the killer sensitivity pattern (KSP) towards a panel of selected killer toxins has proven to be a good auxiliary method. Despite the plethora of studies published, the potential and limitations of the determination of KSPs have never been critically evaluated. In this review the use of this nonmolecular technique as a biotyping tool is discussed and compared with some currently used DNA-based procedures. In addition, methodological, mechanistic and ecological implications are evaluated. PMID- 17425672 TI - An approach to the hypoxic and oxidative stress responses in Kluyveromyces lactis by analysis of mRNA levels. AB - Genome duplication, after the divergence of Saccharomyces cerevisiae from Kluyveromyces lactis along evolution, has been proposed as a mechanism of yeast evolution from strict aerobics, such as Candida albicans, to facultatives/fermentatives, such as S. cerevisiae. This feature, together with the preponderance of respiration and the use of the pentose phosphate pathway in glucose utilization, makes K. lactis a model yeast for studies related to carbon and oxygen metabolism. In this work, and based on the knowledge of the sequence of the genome of K. lactis, obtained by the Genolevures project, we have constructed DNA arrays from K. lactis including a limited amount of selected probes. They are related to the aerobiosis-hypoxia adaptation and to the oxidative stress response, and have been used to test changes in mRNA levels in response to hypoxia and oxidative stress generated by H(2)O(2). The study was carried out in both wild-type and rag2 mutant K. lactis strains in which glycolysis is blocked at the phosphoglucose isomerase step. This approach is the first analysis carried out in K. lactis for the majority of the genes selected. PMID- 17425673 TI - Overproduction of translation elongation factor 1-alpha (eEF1A) suppresses the peroxisome biogenesis defect in a Hansenula polymorpha pex3 mutant via translational read-through. AB - In eukaryotes, elongation factor 1-alpha (eEF1A) is required during the elongation phase of translation. We observed that a portion of the cellular eEF1A colocalizes with purified peroxisomes from the methylotrophic yeast Hansenula polymorpha. We have isolated two genes (TEF1 and TEF2) that encode eEF1A, and which are constitutively expressed. We observed that overproduction of eEF1A suppressed the peroxisome deficient phenotype of an H. polymorpha pex3-1 mutant, which was not observed in a strain deleted for PEX3. The pex3-1 allele contains a UGG to UGA mutation, thereby truncating Pex3p after amino acid 242, suggesting that the suppression effect might be the result of translational read-through. Consistent with this hypothesis, overexpression of the pex3-1 gene itself (including its now untranslated part) partly restored peroxisome biogenesis in a PEX3 null mutant. Subsequent co-overexpression of TEF2 in this strain fully restored its peroxisome biogenesis defect and resulted in the formation of major amounts of full-length Pex3p, presumably via translational read-through. PMID- 17425674 TI - Why do some yeast species require niacin for growth? Different modes of NAD synthesis. AB - NAD holds a key position in metabolism and cellular regulatory events as a major redox carrier and a signalling molecule. NAD biosynthesis pathways have been reconstructed and compared in seven yeast species with completely sequenced genomes, including Saccharomyces cerevisiae, Kluyveromyces lactis, Candida glabrata, Debaryomyces hansenii, Candida albicans, Yarrowia lipolytica and Schizosaccharomyces pombe. Both amino acid and nucleotide sequence similarity analysis in silico indicated that de novo NAD biosynthesis might not exist in K. lactis, C. glabrata and Schiz. pombe, while other species have the kynurenine pathway. It also showed that the NAD salvage pathway via nicotinic acid and nicotinic acid mononucleotide is conserved in all of these yeasts. Deletion of KlNPT1 (the gene for nicotinate phosphoribosyl-transferase) is lethal, which demonstrates that this salvage pathway, utilizing exogenous nicotinic acid, is the unique route to synthesize NAD in K. lactis. The results suggested that the basis of the variation of niacin requirements in yeasts lies in their different combinations of NAD biosynthesis pathways. The de novo pathway is absent but the salvage pathway is conserved in niacin-negative yeasts, while both pathways coexist in niacin-positive yeasts. PMID- 17425675 TI - Mutations in the nucleolar proteins Tma23 and Nop6 suppress the malfunction of the Nep1 protein. AB - The nucleolar Saccharomyces cerevisiae protein Nep1 was previously shown to bind to a specific site of the 18S rRNA and to be involved in assembly of Rps19p into pre-40S ribosome subunits. Here we report on the identification of tma23 and nop6 mutations as recessive suppressors of a nep1(ts) mutant allele and the nep1 deletion as well. Green fluorescent protein fusions localized Tma23p and Nop6p within the nucleolus, indicating their function in ribosome biogenesis. The high lysine content of both proteins and an RNA binding motif in the Nop6p amino acid sequence suggest RNA-binding functions for both factors. Surprisingly, in contrast to Nep1p, Tma23p and Nop6p seem to be specific for fungi as no homologues could be found in higher eukaryotes. In contrast to most other ribosome biogenesis factors, Tma23p and Nop6p are nonessential in S. cerevisiae. Interestingly, the tma23 mutants showed a considerably increased resistance against the aminoglycoside G418, probably due to a structural change in the 40S ribosomal subunit, which could be the result of incorrectly folded 18S rRNA gene, missing rRNA modifications or the lack of a ribosomal protein. PMID- 17425676 TI - The apyrase KlYnd1p of Kluyveromyces lactis affects glycosylation, secretion, and cell wall properties. AB - The Kluyveromyces lactis ORF r_klactIV3,463 on chromosome IV, hereafter named KlYND1, encodes an endoapyrase that has nucleoside phosphatase activity with a lumenal orientation. The enzyme showed equally high activity towards GDP/UDP and ADP, and also showed activity, although to a lesser extent, towards GTP. No activity was detected with the other triphosphates and all monophosphates. The overexpression of KlYND1 in Klgda1Delta cells of K. lactis, devoid of the encoded GDPase/UDPase activity, suppressed the loss of O-glycosylation and cell wall related defects described in such mutants, and suggests a partial overlap of function between the two genes, and therefore some redundancy. The overexpression of KlYND1 in wild-type cells enhanced the secretion of the recombinant human serum albumin and glucoamylase employed as reporters. PMID- 17425677 TI - Intracerebral hemorrhage as initial presentation of gestational choriocarcinoma: a case report and literature review. AB - Choriocarcinoma is the most malignant tumor of gestational trophoblastic neoplasia. It grows rapidly and metastasizes to the lung, liver, and, less frequently, to the brain. One rare case of metastatic cerebral choriocarcinoma with initial presentation of intracerebral hemorrhage is reported. A 40-year-old woman initially presented sudden onset of headache. Intracerebral hemorrhage resulting from ruptured pseudoaneurysm was suspected. Emergent surgery with excision of the pseudoaneurysms was performed. Metastatic choriocarcinoma was accidentally found with positive immunohistochemical staining of cytokeratin and beta subunit of human chorionic gonadotropin (beta-HCG). Choriocarcinoma with brain metastases was diagnosed. She then received chemotherapy with regimen of etoposide, methotrexate, actinomycin-D, cyclophosphamide, and vincristine (EMACO). Elevated serum beta-HCG (30.3 mIU/mL) and new pulmonary lesions were noted by computed tomography 4 months after completion of EMACO. Salvage chemotherapy with etoposide, methotrexate, actinomycin-D, etoposide, and cisplatin (EMAEP) regimen was given. Seven months later after completion of EMAEP, two new pulmonary lesions were detected by positron emission tomography (PET) scan. So she received video-assisted thoracoscopic surgery with tumor excision. Pathologic report confirmed the diagnosis of lung metastases. The patient recovered well. She is free of disease for 12 months. The diagnosis of metastatic cerebral choriocarcinoma was only made by histopathology after craniotomy. Metastatic choriocarcinoma should be always in the differential diagnosis for women at childbearing age presenting with unexplained stroke-like symptoms. In addition, PET scan may be valuable in detecting occult metastatic lesions of choriocarcinoma. PMID- 17425678 TI - Human papillomavirus genotype as a prognostic factor in carcinoma of the uterine cervix. AB - The clinical implications of specific human papillomavirus (HPV) types in invasive cervical carcinomas are only now beginning to be appreciated. The objective of this study was to determine the clinical implications and prognostic value of the HPV genotype in cervical carcinomas. In this study, we employed an HPV DNA chip to detect the type-specific sequence of HPV from cervical swabs taken from women with biopsy-proven neoplastic lesions of the cervix. We divided the patients into four groups: HPV-negative, HPV-16-related, HPV-18-related, and intermediate risk type-related. Associations with clinicopathologic data (stage, histologic type, lymph node status, parametrial invasion, lymphvascular space invasion, tumor size, vaginal involvement) and overall survival were assessed. HPV DNA was detected in 81.4% of the patients, and 19.0% harbored multiple HPV variants. HPV-16-related was the predominant type and was detected in 47.4% (46/97) of the patients. The HPV-16-related types were detected more frequently in patients with squamous cell carcinomas, whereas the HPV-18-related types were more prevalent in cases of adenocarcinomas and adenosquamous carcinomas (P < 0.05). Otherwise, no significant correlations were detected between the HPV genotype and any other clinicopathologic parameters. After a median follow-up of 30 months, the 5-year survival rate was lower in the HPV-18-related patients, but this difference was not found to be statistically significant, according to the results of the log-rank test. We conclude that neither the presence nor type of HPV DNA bears any prognostic significance in cases of cervical carcinoma. PMID- 17425679 TI - Aldo-keto reductase family 1, member B10 in uterine carcinomas: a potential risk factor of recurrence after surgical therapy in cervical cancer. AB - Aldo-keto reductase family 1, member B10 (AKR1B10), an enzyme that converts retinals into retinols is known to detect in non-small cell lung carcinoma (squamous cell- and adeno-carcinomas), but is barely expressed in normal tissues. Since these types of carcinoma occur frequently in the uterus (like in the lung), AKR1B10 may also be overexpressed in two major types of uterine cancer, cervical cancer (CC), and endometrial cancer (EMC). The objective of this study is to investigate AKR1B10 expression in uterine cancer and to analyze its clinical significance. In samples from uterine cancer patients, AKR1B10 was detected in 6 out of 30 (20.0%) CC cases and 6 out of 38 (15.8%) EMC cases. Statistical analysis indicated that AKR1B10 expression was associated with tumor recurrence after surgery and keratinization of squamous cell carcinoma only in CC. Although retinol (a metabolic product by AKR1B10) was observed in the normal epithelium, the molecule was not observed in cancer cells of AKR1B10-positive CC samples suggesting that the recurrence in CC may not depend on the convert of retinals into retinols via AKR1B10, a potential indicator in the management of patients with CC. PMID- 17425680 TI - Bulky lymph node resection in patients with recurrent epithelial ovarian cancer: impact of surgery. AB - The aim of this study was to evaluate the role of systematic lymphadenectomy, feasibility, complications rate, and outcome in epithelial ovarian cancer (EOC) patients with recurrent bulky lymph node disease. A prospective observational study of EOC patients with pelvic/aortic lymph node relapse was conducted between January 1995 and June 2005. After a clinical and laparoscopic staging, secondary cytoreduction, including systematic lymphadenectomy, were performed. The eligibility criteria were as follows: disease-free interval > or =6 months, radiographic finding suggestive of bulky lymph node recurrence, and patients' consent to be treated with chemotherapy. Forty-eight EOC patients with lymph node relapse were recruited. Twenty-nine patients were amenable to cytoreductive surgery. Postoperatively, all patients received adjuvant treatment. The median numbers of resected aortic and pelvic nodes were 15 (2-32) and 17 (8-47), respectively. The median numbers of resected aortic and pelvic positive lymph nodes were 4 (1-18) and 3 (1-17), respectively. The mean size of bulky nodes was 3.3 cm. Four patients (14%) experienced one severe complication. No treatment related deaths were observed. After a median follow-up of 26 months, among cytoreduced patients, 18 women were alive with no evidence of disease, nine were alive with disease. Among the 11 patients not amenable to surgery, five women were alive with persistent disease, six patients died of disease, at a median follow-up of 18 months. Estimated 5-year overall survival and disease-free interval for operated women were 87% and 31%, respectively. In conclusion, patients with bulky lymph node relapse can benefit from systematic lymphadenectomy in terms of survival. The procedure is feasible with an acceptable morbidity rate. PMID- 17425681 TI - Survival and prognostic factors in early-stage epithelial ovarian carcinoma treated with taxane-based adjuvant chemotherapy. AB - The present study was undertaken with the question about the outcome (recurrence free survival, [RFS]) after adjuvant chemotherapy with taxane and carboplatin in the early stages of epithelial ovarian cancer after primary surgery. Treatment related toxicity was also evaluated. A total of 113 patients were included in this study. The 5-year RFS rate for all 113 patients treated with adjuvant chemotherapy including taxane and carboplatin after primary surgery was 79%. The 5-year RFS rate for 85 patients in FIGO stage I was 85% and for 18 patients in FIGO stage II, it was 44%. For clear-cell carcinomas, the RFS was 87%. In univariate analysis, recurrent disease was associated with both FIGO stage and tumor grade, but in multivariate logistic regression analysis of prognostic factors for tumor recurrences, only FIGO stage (stage I versus stage II) was a significant and independent prognostic factor. However, an odds ratio (OR) of 1.9 for tumor grade (grade 3 versus grades 1-2) demonstrated two times increased risk for recurrence in a patient with a grade 3 tumor compared with grade 1-2 tumors. Furthermore, an OR of 0.39 for lymph node sampling versus no sampling meant 61% reduced risk for recurrence for a patient who had undergone lymph node sampling at surgical staging laparotomy. The major toxicities in the present study were myelosuppression (46%) and neurotoxicity (34%). Despite the use of prophylaxis, severe paclitaxel-related hypersensitivity occurred in three patients (3%). PMID- 17425682 TI - Overexpressed active Notch1 induces cell growth arrest of HeLa cervical carcinoma cells. AB - Human cervical carcinoma is one of the most common malignant tumors, but the mechanisms that orchestrate the multiple oncogenic insults required for initiation and progression are not clear. Notch signaling plays a critical role in maintaining the balance between cell proliferation, differentiation, and apoptosis, but perturbed Notch signaling may contribute to tumorigenesis. We now show that Notch1 is detected in all cervical cancer, including advanced diseases. We also constitutively overexpressed active Notch1 in human cervical carcinoma to explore the effects of Notch1 signaling on human cervical carcinoma cell growth and to investigate the underlying molecular mechanisms. The signaling may participate in the development of human cervical carcinoma cells, but overexpressed active Notch1 inhibits their growth through induction of cell cycle arrest. Increased Notch1 signaling induced a downmodulation of human papillomavirus transcription through suppression of activator protein (AP)-1 activity by upregulation of c-Jun and the decreased expression of c-Fos. Thus, Notch1 signaling plays a key role and exerts dual effects, functioning in context specific manner. PMID- 17425683 TI - Advanced stage of cervical carcinoma undiagnosed during antenatal period in term pregnancy and concomitant metastasis on episiotomy scar during delivery: a case report and review of the literature. AB - The objective of this study is to review the implantation of malignant cells of cervical cancer in an episiotomy site. This is the second case of cervical cancer with concomitant episiotomy metastasis in the literature. The treatment consisted of radiochemotherapy. There was no confirmed recurrent disease after 1 year of follow-up in our reported case. In conclusion, pregnancy appears to be a chance for cervical cancer screening as a part of prenatal care. If cervical cancer is diagnosed in a woman after vaginal delivery, she must be examined carefully including the episiotomy site. PMID- 17425684 TI - The relation between age, time since menopause, and endometrial cancer in women with postmenopausal bleeding. AB - The objective is to assess among women with postmenopausal bleeding the relationship of age and time since menopause on one hand and the presence of endometrial cancer and atypical hyperplasia on the other hand. In a multicenter prospective cohort study, 614 women presenting with postmenopausal bleeding were included. Women underwent transvaginal sonography and, in cases where the endometrial thickness was >4 mm, endometrial sampling. Splines were used to assess the association between each of the continuous variables and (pre)malignancy of the endometrium. Subsequently, univariate and multivariate analysis were performed. The average age for women without (pre)malignancy was 61.7 years (SD 9.8). As malignant and premalignant cases were found to have similar age, these subgroups were merged in the analyses. Age was an independent predictor of (pre)malignancy. In women younger than 55 years, the odds ratio was 1.9 (95% CI: 1.1-3.3) for each year under 55 years of age and 1.03 (95% CI: 1.00 1.06) for each year over 55 years of age. The risk of (pre)malignancy of the endometrium was 4.9% in women less than 3 years postmenopausal versus 19.7% in women more than 20 years postmenopausal. However, in a multivariate analysis only age contributed to the prediction of risk. This study demonstrates that, in postmenopausal women with vaginal bleeding, the risk of (pre)malignancy of the endometrium is low in women under 50 years of age, increases considerably until 55 years of age, and rises only modestly with further advancing age. Future studies should explore whether these findings can be incorporated in the diagnostic work-up of women with postmenopausal bleeding. PMID- 17425685 TI - von Willebrand factor A1 domain: stuck in the middle. PMID- 17425686 TI - Clearance mechanisms of von Willebrand factor and factor VIII. PMID- 17425687 TI - Association between asymptomatic deep vein thrombosis detected by venography and symptomatic venous thromboembolism in patients undergoing elective hip or knee surgery. AB - BACKGROUND: Venography is commonly used to compare the efficacy of different thromboprophylaxis strategies for preventing deep vein thrombosis (DVT) in patients undergoing total hip replacement (THR) or total knee replacement (TKR). METHODS: We explored the relation between asymptomatic DVT and symptomatic venous thromboembolism (VTE) in patients undergoing THR or TKR treated with standard doses of enoxaparin (30 mg b.i.d. or 40 mg o.d.) by comparing the incidence of asymptomatic DVT in venographic studies with the incidence of symptomatic VTE in studies where venography was not performed. RESULTS: In 10 venographic studies involving 5796 patients, the incidence of asymptomatic DVT after THR was 13.2% [95% CI, 12.2-14.2%] and after TKR was 38.1% (95% CI, 35.5-40.8%). In two studies involving 3500 patients who did not undergo venography, the 90-day incidence of symptomatic VTE after THR was 2.7% (95% CI, 2.1-3.4%) and after TKR was 1.8% (95% CI, 0.9-2.7%). For every symptomatic VTE in THR studies where venography was not performed there were five asymptomatic DVTs in the venographic studies; for TKR, the ratio was 1:21. The incidence of asymptomatic DVT and the symptomatic VTE/asymptomatic DVT ratio was influenced by the venogram reading committee (Gothenburg vs. Hamilton: total DVT after THR, 19.5% vs. 8.7%, P < 0.0001; for TKR, 42.7% vs. 27.2%, P < 0.0001). CONCLUSIONS: Comparisons across trials show a consistent relation between asymptomatic venographic DVT in patients undergoing elective THR or TKR surgery and symptomatic VTE in patients not undergoing venography. Differences exist in the strength of the relation depending on the type of surgery and the venogram reading committee. PMID- 17425688 TI - Establishment of the enzyme-linked immunosorbent assay system to detect the amino terminal secretory form of rat Erc/Mesothelin. AB - By representational difference analysis, we previously identified the rat Erc (Expressed in renal carcinoma) gene that was more abundantly expressed in the renal carcinoma tissues of Eker rats than in the rat normal kidney. In this study, we raised antibodies against the amino-terminal portion of the rat Erc, and demonstrated the existence of a approximately 30-kDa secretory form in the supernatant of cultured cells derived from rat renal carcinoma. The enzyme-linked immunosorbent assay (ELISA) system using these antibodies detected high concentrations of this form in the sera of Eker rats bearing renal carcinomas, and in the sera of rats transplanted with mesothelioma cells. Mesothelin, a human homolog of the rat Erc, was recently reported to be a serum marker of malignant mesothelioma. The prognosis of mesothelioma is poor and there is no effective treatment at present. There are several rat model systems of mesothelioma that may be promising tools in the development of an antimesothelioma treatment. We hope our ELISA to detect the soluble form of rat Erc/Mesothelin is useful in the rat model system to exploit the antimesothelioma therapy to be used in human cases. PMID- 17425689 TI - Rapamycin, a specific inhibitor of the mammalian target of rapamycin, suppresses lymphangiogenesis and lymphatic metastasis. AB - Tumor lymphangiogenesis is now known to play a causal role in lymph node metastasis, and thus its inhibition would have great significance for the prevention of lymph node metastasis in cancer therapy. VEGF-C has recently been identified as a key molecule that involved in tumor lymphangiogenesis and lymphatic metastasis. However, the expressional regulation of VEGF-C is not fully understood. We investigated the role of mTOR, which is a downstream kinase of the phosphatidylinositol 3-kinase/Akt pathway, and the MAPK family (MEK1/2, p38, and JNK) in the regulation of VEGF-C and VEGF-A expression in B13LM cells, a lymphatic metastasis-prone pancreatic tumor cell line. We also investigated the antilymphangiogenic effect of rapamycin, a specific inhibitor of mTOR in vivo using male BALB/c nu/nu mice. VEGF-C expression was inhibited by the inhibitors for mTOR, p38, and JNK, but not by the inhibitor for MEK1/2, whereas VEGF-A expression was inhibited by all four of these inhibitors. The serum starvation induced expression of VEGF-C was inhibited by rapamycin, whereas that of VEGF-A was incompletely inhibited. The metastatic experiment in vivo demonstrated that the number and the area of lymphatic vessels in the primary tumors were significantly decreased by rapamycin. Finally, the lymph node metastasis was significantly suppressed in rapamycin-treated mice. Our results suggest that mTOR, p38, and JNK play important roles in VEGF-C expression, and that rapamycin has an antilymphangiogentic effect and exerts the expected inhibition of lymphatic metastasis. PMID- 17425690 TI - Photodynamic therapy-generated tumor cell lysates with CpG-oligodeoxynucleotide enhance immunotherapy efficacy in human papillomavirus 16 (E6/E7) immortalized tumor cells. AB - Immunotherapy with photodynamic therapy (PDT) offers great promise as a new alternative for cancer treatment; however, its use remains experimental. In this study, we examined the immunotherapeutic significance of human papillomavirus (HPV)-immortalized tumor cell lysates induced by PDT with CpG oligodeoxynucleotide (ODN). PDT-cell lysates were generated by irradiating Radachlorin (5 microg/mL) preloaded TC-1 cells carrying HPV 16 E7. PDT-cell lysates plus ODN coinjection for protection against E7-expressing tumors as well as specific immune responses were evaluated with the following tests: heat shock protein 70 (HSP70) enzyme-linked immunosorbent assay, in vitro and in vivo tumor growth inhibition, interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) assay, cytotoxic T-lymphocyte assay, and fluorescence activated cell sorting (FACS) analysis. PDT-cell lysates plus ODN coinjection showed a significant suppression of tumor growth at both prophylactic and therapeutic levels, compared to PDT (or F/T)-cell lysates or ODN alone. In addition, we evaluated the level of the immune response with the coinjection. HSP70, an important regulator of inflammatory and immune response, was observed in abundance in the PDT-cell lysates. IFN-gamma production and cytotoxic T lymphocytes (CTL) responses were induced by PDT-cell lysates plus ODN injection. The coinjection resulted in PDT-cell lysate-specific antibodies (IgG1, IgG2a, IgG2b, and IgG3) and T-helper cell responses significantly higher than PDT-cell lysates alone. Moreover, IFN-gamma production and CTL responses were significantly induced in the PDT-cell lysate plus ODN immunized groups. These enhanced immune responses appeared to be mediated by CD8+ T cells only. These data suggest that PDT-cell lysates plus ODN injection may be an effective approach to induce CTL immune responses as a possible immunotherapeutic strategy for cancer therapy. PMID- 17425693 TI - Abstracts of the 13th Annual Conference of the British HIV Association with the British Infection Society. Edinburgh, United Kingdom. April 25-28, 2007. PMID- 17425694 TI - Reduced tear production in three canine endocrinopathies. AB - OBJECTIVES: Previous reports have suggested that hypothyroid and diabetic patients can be predisposed to keratoconjunctivitis sicca. This study aimed to measure tear production in dogs with diabetes, hypothyroidism and hyperadrenocorticism using the Schirmer tear test and to compare these results with Schirmer tear test values for a group of normal dogs. METHODS: Schirmer tear tests were performed on 16 dogs with hyperadrenocorticism, 18 with diabetes and 12 with hypothyroidism together with 100 control dogs. Corneal sensitivity was also measured in 12 of the 18 diabetic dogs with a Cochet Bonnet aesthesiometer and compared with age- and breed-matched normal dogs. RESULTS: Schirmer tear test values in dogs with hypothyroidism, hyperadrenocorticism and diabetes were 12.3+/ 3.2, 14.0+/-4.0 and 12.3+/-5.3 mm/minutes, respectively. Schirmer tear test values were significantly lower than that for the control group (19.6+/-4.2 mm/minutes) in all dogs with an endocrinopathy. Only in two hypothyroid dogs and three diabetics, this was manifested as profound keratoconjunctivitis sicca with Schirmer tear test value lower than 5 mm/minutes. Diabetic dogs had significantly reduced corneal sensitivity compared with a matched set of control dogs. CLINICAL SIGNIFICANCE: This study shows a significant reduction in tear production in animals with diabetes mellitus, hypothyroidism and hyperadrenocorticism. Further research is needed to elucidate the mechanisms by which this reduction in tear production occurs. Assessment of tear production should be undertaken in animals diagnosed with these endocrinopathies, as these animals may progress to clinical keratoconjunctivitis sicca. PMID- 17425695 TI - Antibiotic susceptibility of bacterial isolates from corneal ulcers of dogs in Taiwan. AB - OBJECTIVES: To analyse antibiotic susceptibility of bacteria associated with ulcerative keratitis in dogs. METHODS: Bacteria isolated from 190 eyes with ulcerative keratitis were identified, and the antibiotic susceptibility of isolates was studied. RESULTS: In total, 258 species of bacteria were isolated from the 190 eyes. Of the isolates, 78 per cent were Gram-positive and 28 per cent were Gram-negative bacteria. The most commonly isolated Gram-positive bacteria in dogs were Staphylococcus spp (49 per cent), Streptococcus spp (7 per cent) and Corynebacterium spp (7 per cent); while Pseudomonas aeruginosa (7.6 per cent) and Escherichia coli (5.8 per cent) were the commonest Gram-negative pathogens. Resistance to commonly used ophthalmic antibiotics was seen in Staphylococcus, Streptococcus, Corynebacterium, Pseudomonas and Escherichia species isolates. CLINICAL SIGNIFICANCE: Isolates from dogs with corneal ulcers in Taiwan may be resistant to several commonly used ophthalmic preparations. Ciprofloxacin showed good action against most isolates, with the notable exception of Streptococcus species. Chloramphenicol or cephalothin had the best in vitro action against the Streptococcus species isolates. PMID- 17425696 TI - Solitary extramedullary plasmacytoma of the canine larynx. AB - An 11-year-old, female, spayed cocker spaniel was presented with dysphonia caused by a solitary laryngeal mass. Excisional biopsy was performed, and a diagnosis of plasmacytoma was made on the basis of histological examination. Further investigations showed no signs of systemic involvement. Coarse fractionated radiation therapy failed to control the tumour. Therapy was successfully instituted with a conventional combination chemotherapy protocol over a period of 14 months. The dog remains disease free 30 months after diagnosis. Most solitary, extramedullary plasmacytomas in dogs arise in the gastrointestinal tract, with fewer reports in other sites. The larynx is an uncommon sight of involvement in any species, and to the authors' knowledge, this is the first report of this tumour type in the canine larynx. In contrast to the therapeutic benefits reported in humans, the combination of surgery and radiation therapy was unsuccessful in this case, although sustained remission was gained following chemotherapy. PMID- 17425697 TI - Five cases of canine dysautonomia in England (2004 to 2006). AB - Canine dysautonomia was diagnosed definitively in five dogs by histopathology. All dogs were seen between June 2004 and July 2006 and originated from south-east England; four dogs originated from an urban area and one from a rural area. Of the urban dogs, one had recently visited Scotland and one had visited a kennel in a rural area nearby. Acute-onset but progressive vomiting, diarrhoea, depression and inappetence were the most common presenting clinical signs. Reduced or absent anal tone, dysuria, absence of pupillary light reflexes with intact vision, mydriasis, decreased corneal sensitivity and nictitating membrane protrusion were among the most frequent neurological findings. Abnormalities in pharmacological autonomic and physiological function testing (including orthostatic hypotension in two dogs) and diagnostic imaging studies were detected in some of the animals. All dogs failed to respond adequately to treatment, and given the poor prognosis, they were eventually euthanased. Histopathology identified marked chromatolysis of ganglion cell bodies. This case series emphasises that dysautonomia should be considered when a dog is presented in the UK with acute- or subacute-onset gastrointestinal signs and compatible physical and neurological abnormalities. PMID- 17425698 TI - Aberrant extradural spinal migration of Spirocerca lupi: four dogs. AB - Spirocerca lupi is a nematode mainly affecting dogs but has been found in other animals, particularly carnivores. Anatomical areas of typical and aberrant migration vary. This report describes four cases of Spirocerca lupi causing neurological symptoms, similar to thoracolumbar disc syndrome, as a result of aberrant migration of the nematode into the spinal canal. In two of the cases, the nematode could be demonstrated intraoperatively. The third was discovered on post-mortem examination, while the fourth case displayed compelling evidence of Spirocerca lupi involvement. Surgical removal of the Spirocerca lupi nematode would be the treatment of choice in cases of spinal migration, with therapeutic and preventive treatment with doramectin instituted to treat remote sites and prevent re-infection. In areas endemic for spirocercosis, Spirocerca lupi should be considered as an important differential diagnosis in cases that are presented with clinical signs suggestive of a spinal cord lesion. PMID- 17425699 TI - Atrial myocarditis as a cause of sinus arrest in a dog. AB - A 10-year-old male cairn terrier cross was presented with a history of myxomatous mitral valve disease diagnosed six months previously and with a four-week history of intermittent collapse. On 24 hour electrocardiograph (Holter) analysis, periods of no discernable electrical cardiac activity, which coincided with three collapsing episodes, were identified. Unfortunately, on re-presentation for removal of the Holter monitor, the dog collapsed and died. A post-mortem examination was conducted, and histology of the right and left atrium showed evidence of myocarditis. This is the first reported case, to our knowledge, of collapse because of electrical asystole in a dog with atrial myocarditis. PMID- 17425700 TI - Aortic endocarditis associated with a perforated septal membranous aneurysm in a boxer dog. AB - Perimembranous ventricular septal defect is a common congenital heart disease in the dog. It can partially or completely close with age by development of a membranous ventricular septal aneurysm. Aortic endocarditis is a reported complication of ventricular septal defect and membranous ventricular septal aneurysm in human beings. This report describes a case of aortic endocarditis associated with a membranous ventricular septal aneurysm perforated by a small ventricular septal defect in a boxer dog. PMID- 17425701 TI - Necrotising encephalitis in a French bulldog. AB - A 20-month-old, female French bulldog was presented with a three-month history of generalised seizures and progressive ataxia with occasional falling over on either side. Neurological examination revealed signs, suggesting a multifocal intracranial lesion. Magnetic resonance imaging of the brain revealed two connected lesions on the left side of the caudal brainstem and a further lesion in the cerebrum. The dog was euthanased, and histopathological examination revealed lesions which closely resembled those of necrotising encephalitis in Yorkshire terriers. This is the first case describing this type of necrotising encephalitis in a French bulldog. PMID- 17425702 TI - A new surgical method for the control of the epiphora in dogs: modified parotid duct transposition. AB - A five-year-old, female, spayed beagle with a history of unilateral epiphora of several months' duration as a result of inadequacy of the lacrimal drainage system was treated successfully with a new surgical method of parotid duct transposition. Data from complete preoperative ophthalmological examination, surgical technique, postoperative treatment, contrast radiography and histological examination were recorded. Absence of complications and complete patency of the stoma at 30 days after surgery confirmed the validity of this new technique. The use of an anatomic duct may be advantageous compared with the current techniques of conjunctivorhinostomy, conjuntivobuccostomy and conjunctival maxillary sinusostomy for the treatment of epiphora in the dogs caused by inadequacy of the lacrimal drainage system. PMID- 17425703 TI - Migraine: theory to therapy--25 years of progress. PMID- 17425704 TI - The discovery of a new drug class for the acute treatment of migraine. AB - The history of the scientific ideas and events that led to the discovery of sumatriptan is outlined with personal reminiscences about individuals who influenced the approach. The development of sumatriptan revolutionized the acute treatment of migraine and led to the availability of a number of other triptans. The anti-migraine effects of all the triptans are mediated via 5-HT(1B), and possibly 5-HT(1D) receptors, which transduce their effects via G; proteins. This suggests that agonists at other G(i) protein-coupled receptor types appropriately located (eg, somatostatin sst(2), adenosine A(1)) should be examined for their effects on the trigeminovascular system, Studies on such receptor targets may provide insight into a novel approach towards the design of new anti-migraine drugs. PMID- 17425705 TI - Ten lessons on the epidemiology of migraine. AB - The epidemiologic studies of the last 15 years have taught many lessons that will help define the clinical and research agenda in migraine for the next 15 years. Because the condition is common, disabling, and costly (Lesson 1) and because it has major comorbidities (Lesson 2), migraine is an important target for treatment. Because consultation, diagnosis, and effective treatment rates are low (Lessons 3 and 4) it is important to develop strategies for closing the gap between treatment available and treatment delivered in the primary care setting where the majority of patients seek care (Lesson 5 and 6). Strategies include public and provider education as well as the use of screening tools such as ID migraine to improve the recognition of migraine in primary care (Lesson 7). In addition, strategies that measure disability, with tools such as MIgraine Disability ASssessment, when coupled with treatment guidelines which consider illness severity in the selection of treatment provide great hope for improving patient outcomes (Lesson 8). Finally, the emerging evidence that migraine is progressive in a subgroup (Lesson 9) mandates the development of strategies to reduce the risk of progression by addressing modifiable risk factors and by assessing the effects of treatment on headache progression (Lesson 10). PMID- 17425706 TI - Major therapeutic advances in the past 25 years. PMID- 17425707 TI - Stratification of headache care. PMID- 17425708 TI - New migraine and pain research. AB - An understanding of the pathophysiology and pharmacology of migraine has been driven by astute clinical observations, elegant experimental medicine studies and importantly by the introduction of highly effective selective anti-migraine agents such as the Triptan 5-HT(1B/1D) agonists. New investigational migraine therapies such CGRP antagonists target key components of the trigeminal sensory neuroinflammatory response and show promise for the future. Cutting edge molecular profiling studies looking at gene expression during chronic pain are now being used to reveal the cell biology of pain and new potential therapeutic targets. Translational neuroimaging research can link the laboratory and the clinic and is now being used to help understand the neural systems biology of migraine. Research into migraine has generated sophisticated hypotheses that encompass primary CNS dysfunction, trigeminovascular activation, pain perception and activation of associated neural circuits involved in affective functions providing a rich framework within which to design and test future migraine treatment strategies. PMID- 17425709 TI - The convergence hypothesis. AB - The Convergence Hypothesis postulates a single pathophysiological mechanism to explain the clinical spectrum of primary headaches seen in patients with migraine. The history and the scientific underpinnings of the Convergence Hypothesis are presented. Extrapolations from the Convergence Hypothesis are used to explore the evolution of episodic to chronic migraine and the development of common migraine co-morbidities as a consequence of frequent migraine. A patient staging system is presented to illustrate this transformation process in migraine patients. PMID- 17425710 TI - Current trends in migraine prophylaxis. AB - A variety of drugs from diverse pharmacological classes are in use for migraine prevention. Traditionally, they have been discovered by serendipity. Examples include beta-adrenergic blockers, anticonvulsants, tricyclic antidepressants, and serotonin receptor antagonists. The mechanisms of action of migraine preventive drugs are multiple but it is postulated that they converge on two targets: (1) inhibition of cortical excitation; (2) restoring nociceptive dysmodulation. The antiepileptic drugs (e.g., topiramate, valproate, gabapentin), calcium channel blockers such as verapamil, and inhibitors of cortical spreading depression are some examples of drugs that reduce neuronal hyperexcitability. On the other hand, modulators of the serotonergic and adrenergic systems and cholinergic enhancing drugs may restore descending nociceptive inhibition and play a role in migraine prevention. To date, Level 1 evidence and clinical experience favor the use of the antidepressant amitriptyline, the anticonvulsants divalproex and topiramate, and the beta-adrenergic blockers propranolol, timolol and metoprolol as first line migraine preventive drugs. The evidence for others (e.g., verapamil) is not as strong. Migraine preventive drugs have varying degrees of adverse effects, some of which could be limiting, and their efficacy should balanced with their risks of adverse effects, patients' expectations and desires, and compliance. It is hoped that future migraine preventive drugs target migraine mechanisms more specifically, which could well enhance the therapeutic index. PMID- 17425711 TI - Pathophysiology of headache--past and present. PMID- 17425712 TI - Homogalacturonan synthesis in Arabidopsis thaliana requires a Golgi-localized protein with a putative methyltransferase domain. AB - Pectins are a family of complex cell-wall polysaccharides, the biosynthesis of which remains poorly understood. We identified dwarf mutants with reduced cell adhesion at a novel locus, QUASIMODO2 (QUA2). qua2-1 showed a 50% reduction in homogalacturonan (HG) content compared with the wild type, without affecting other cell-wall polysaccharides. The remaining HG in qua2-1 showed an unaltered degree of methylesterification. Positional cloning and GFP fusions showed that QUA2, consistent with a role in HG synthesis, encodes a Golgi-localized protein. In contrast to QUA1, another Golgi-localized protein required for HG-synthesis, QUA2 does not show sequence similarity to glycosyltransferases, but instead contains a putative methyltransferase (MT) domain. The Arabidopsis genome encodes 29 QUA2-related proteins. Interestingly, the transcript profiles of QUA1 and QUA2 are correlated and other pairs of QUA1 and QUA2 homologues with correlated transcript profiles can be identified. Together, the results lead to the hypothesis that QUA2 is a pectin MT, and that polymerization and methylation of homogalacturonan are interdependent reactions. PMID- 17425713 TI - Orthologous comparison in a gene-rich region among grasses reveals stability in the sugarcane polyploid genome. AB - Modern sugarcane (Saccharum spp.) is an important grass that contributes 60% of the raw sugar produced worldwide and has a high biofuel production potential. It was created about a century ago through hybridization of two highly polyploid species, namely S. officinarum and S. spontaneum. We investigated genome dynamics in this highly polyploid context by analyzing two homoeologous sequences (97 and 126 kb) in a region that has already been studied in several cereals. Our findings indicate that the two Saccharum species diverged 1.5-2 million years ago from one another and 8-9 million years ago from sorghum. The two sugarcane homoeologous haplotypes show perfect colinearity as well as high gene structure conservation. Apart from the insertion of a few retrotransposable elements, high homology was also observed for the non-transcribed regions. Relative to sorghum, the sugarcane sequences displayed colinearity, with the exception of two genes present only in sorghum, and striking homology in most non-coding parts of the genome. The gene distribution highlighted high synteny and colinearity with rice, and partial colinearity with each homoeologous maize region, which became perfect when the sequences were combined. The haplotypes observed in sugarcane may thus closely represent the ancestral Andropogoneae haplotype. This analysis of sugarcane haplotype organization at the sequence level suggests that the high ploidy in sugarcane did not induce generalized reshaping of its genome, thus challenging the idea that polyploidy quickly induces generalized rearrangement of genomes. These results also confirm the view that sorghum is the model of choice for sugarcane. PMID- 17425714 TI - Plant cells must pass a K+ threshold to re-enter the cell cycle. AB - Potassium is an inevitable component of plant life, and potassium channels play a pivotal role in plant growth and development. The role of potassium and of K(+) channels in plant cell division and cell-cycle progression, however, has not been determined so far. K(+) channel blocker studies with synchronized tobacco BY-2 cells revealed that K(+) uptake is required for proper cell-cycle progression during the transition from G(1) to S phase. Electrophysiological studies (patch clamp and voltage-clamp techniques) showed a cell-cycle dependency of K(+) channel activities and reduced driving force for K(+) uptake in dividing cells. Among the four Shaker-like K(+) channel genes expressed in BY-2 cells, NKT1 represents an inwardly rectifying K(+) channel that mediates K(+) uptake. NKT1 is transcriptionally induced during G(1) phase, while transcripts of the outward rectifier NTORK1 dominate S phase. Elongating BY-2 cells appeared hyperpolarized (-101 +/- 11 mV), and had elevated osmotic pressure and approximately twice the turgor pressure when compared with depolarized (-64 +/- 8 mV) dividing cells. This indicates that cells have to gain a threshold K(+) level to re-enter the cell cycle. Based on these findings, turgor regulation through modulation of K(+) channel density in plant cell division and cell-cycle progression is discussed. PMID- 17425715 TI - Self-compatibility in Brassica napus is caused by independent mutations in S locus genes. AB - Brassica napus is an amphidiploid species with the A genome from Brassica rapa and the C genome from Brassica oleracea. Although B. rapa, B. oleracea and artificially synthesized amphidiploids with the AC genome are self-incompatible, B. napus is self-compatible. Six S genotypes were identified in B. napus, five of which had class I S haplotypes from one species and a class II S haplotype from the other species, and mutations causing self-compatibility were identified in three of these S genotypes. The most predominant S genotype (BnS-1;BnS-6), which is that of cv. 'Westar', had a class I S haplotype similar to B. rapa S-47 (BrS 47) and a class II S haplotype similar to B. oleracea S-15 (BoS-15). The stigmas of 'Westar' rejected the pollen grains of both BrS-47 and BoS-15, while reciprocal crossings were compatible. Insertion of a DNA fragment of about 3.6 kb was found in the promoter region of the SP11/SCR allele of BnS-1, and transcripts of SP11/SCR were not detected in 'Westar'. The nucleotide sequence of the SP11 genomic DNA of BnS-6 was 100% identical to that of SP11 of BoS-15. Class I SP11 alleles from one species showed dominance over class II SP11 alleles from the other species in artificially synthesized B. napus lines, suggesting that the non functional dominant SP11 allele suppressed the expression of the recessive SP11 allele in 'Westar'. Two other S genotypes in B. napus also had non-functional class I S haplotypes together with recessive BnS-6. These observations suggest independent origins of self-compatibility in B. napus. PMID- 17425716 TI - Arabidopsis thaliana plants possess a specific farnesylcysteine lyase that is involved in detoxification and recycling of farnesylcysteine. AB - In plants, prenylated proteins are involved in actin organization, calcium mediated signal transduction, and many other biological processes. Arabidopsis thaliana mutants lacking functional protein prenyltransferase genes have also revealed roles for prenylated proteins in phytohormone signaling and meristem development. However, to date, the turnover of prenylated plant proteins and the fate of the prenylcysteine (PC) residue have not been described. We have detected an enzyme activity in Arabidopsis plants that metabolizes farnesylcysteine (FC) to farnesal, which is subsequently reduced to farnesol. Unlike its mammalian ortholog, Arabidopsis FC lyase exhibits specificity for FC over geranylgeranylcysteine (GGC), and recognizes N-acetyl-FC (AFC). FC lyase is encoded by a gene on chromosome 5 of the Arabidopsis genome (FCLY, At5g63910) and is ubiquitously expressed in Arabidopsis tissues and organs. Furthermore, T-DNA insertions into the FCLY gene cause significant decreases in FC lyase activity and an enhanced response to abscisic acid (ABA) in seed germination assays. The effects of FCLY mutations on ABA sensitivity are even greater in the presence of exogenous FC. These data suggest that plants possess a specific FC detoxification and recycling pathway. PMID- 17425717 TI - A simplified method for the analysis of transcription factor-promoter interactions that allows high-throughput data generation. AB - Transient expression systems are intensively used to study the transactivation potential of transcription factors and to confirm target promoters. Here we present a novel system based on the high-efficiency transformation of cultured Arabidopsis thaliana cells by agrobacteria. To demonstrate the potential of this system, we compared it with a commonly used protoplast transfection assay, and studied the regulation of phenylpropanoid biosynthetic pathway genes by various transcription factors. Both systems led to comparable results on the regulation of the promoters tested. However, the agrobacterium-mediated co-transformation assay needs significantly less time, requires only mixing of cultured plant cells with agrobacteria, is less labour-intensive and allows handling of multiple assays in parallel, making it suitable for medium- to high-throughput analyses. In addition, the binary vectors used are the same for both cell-based assays and stable plant transformations. PMID- 17425718 TI - From seedling to mature plant: arabidopsis plastidial genome copy number, RNA accumulation and transcription are differentially regulated during leaf development. AB - Little is known about DNA and RNA metabolism during leaf development and aging in the model organism Arabidopsis. Therefore we examined the nuclear and plastidial DNA content of tissue ranging in age from 2-day-old cotyledons to 37-day-old senescent rosette leaves. Flow-cytometric analysis showed an increase in nuclear DNA ploidy levels of up to 128 genome copies per nucleus in older leaves. The copy numbers of nuclear 18S-rRNA genes were determined to be 700 +/- 60 per haploid genome. Adjusted to the average level of nuclear DNA polyploidism per cell, plastome copy numbers varied from about 1000 to 1700 per cell without significant variation during development from young to old rosette leaves. The transcription activity of all studied plastid genes was significantly reduced in older rosette leaves in comparison to that in young leaves. In contrast, levels of plastidial transcript accumulation showed different patterns. In the case of psbA, transcripts accumulated to even higher levels in older leaves, indicating that differential regulation of plastidial gene expression occurs during leaf development. Examination of promoter activity from clpP and rrn16 genes by primer extension analyses revealed that two RNA polymerases (NEP and PEP) transcribe these genes in cotyledons as well as in young and senescent leaves. However, PEP may have a more prominent role in older rosette leaves than in young cotyledons. We conclude that in cotyledons or leaves of different ages plastidial gene expression is regulated at the transcriptional and post-transcriptional levels, but not by plastome copy number. PMID- 17425719 TI - Sulfite oxidase protects plants against sulfur dioxide toxicity. AB - The gaseous pollutant SO(2) readily reacts with water to form sulfite that impacts deleteriously on animal and plant health. By modulating the level of sulfite oxidase (SO) that catalyzes the transformation of sulfites to the non toxic sulfate, we show that Arabidopsis and tomato plants can be rendered resistant or susceptible to SO(2)/sulfite. Plants in which sulfite oxidase expression was abrogated by RNA interference (RNAi) accumulated relatively less sulfate after SO(2) application and showed enhanced induction of senescence and wounding-associated transcripts, leaf necrosis and chlorophyll bleaching. In contrast, SO overexpression lines accumulated relatively more sulfate and showed little or no necrosis after SO(2) application. The transcript of sulfite reductase, a chloroplast-localized enzyme that reduces sulfites to sulfides, was shown to be rapidly induced by SO(2) in a sulfite oxidase-dependent manner. Transcripts of other sulfite-requiring enzymatic activities such as mercaptopyruvate sulfur transferases and UDP-sulfoquinovose synthase 1 were induced later and to a lesser extent, whereas SO was constitutively expressed and was not significantly induced by SO(2). The results imply that plants can utilize sulfite oxidase in a sulfite oxidative pathway to cope with sulfite overflow. PMID- 17425720 TI - Convergent evolution in the BAHD family of acyl transferases: identification and characterization of anthocyanin acyl transferases from Arabidopsis thaliana. AB - Members of the BAHD family of plant acyl transferases are very versatile catalytically, and are thought to be able to evolve new substrate specificities rapidly. Acylation of anthocyanins occurs in many plant species and affects anthocyanin stability and light absorption in solution. The versatility of BAHD acyl transferases makes it difficult to identify genes encoding enzymes with defined substrate specificities on the basis of structural homology to genes of known catalytic function alone. Consequently, we have used a modification to standard functional genomics strategies, incorporating co-expression profiling with anthocyanin accumulation, to identify genes encoding three anthocyanin acyl transferases from Arabidopsis thaliana. We show that the activities of these enzymes influence the stability of anthocyanins at neutral pH, and some acylations also affect the anthocyanin absorption maxima. These properties make the BAHD acyl transferases suitable tools for engineering anthocyanins for an improved range of biotechnological applications. PMID- 17425721 TI - Altered expression of plant lysyl tRNA synthetase promotes tRNA misacylation and translational recoding of lysine. AB - The Arabidopsis thaliana lysyl tRNA synthetase (AtKRS) structurally and functionally resembles the well-characterized prokaryotic class IIb KRS, including the propensity to aminoacylate tRNA(Lys) with suboptimal identity elements, as well as non-cognate tRNAs. Transient expression of AtKRS in carrot cells promotes aminoacylation of such tRNAs in vivo and translational recoding of lysine at nonsense codons. Stable expression of AtKRS in Zea mays causes translational recoding of lysine into zeins, significantly enriching the lysine content of grain. PMID- 17425722 TI - The maize (Zea mays L.) RTCS gene encodes a LOB domain protein that is a key regulator of embryonic seminal and post-embryonic shoot-borne root initiation. AB - Maize has a complex root system composed of different root types formed during different stages of development. The rtcs (rootless concerning crown and seminal roots) mutant is impaired in the initiation of the embryonic seminal roots and the post-embryonic shoot-borne root system. The primary root of the mutant shows a reduced gravitropic response, while its elongation, lateral root density and reaction to exogenously applied auxin is not affected. We report here the map based cloning of the RTCS gene which encodes a 25.5 kDa LOB domain protein located on chromosome 1S. The RTCS gene has been duplicated during evolution. The RTCS-LIKE (RTCL) gene displays 72% sequence identity on the protein level. Both genes are preferentially expressed in roots. Expression of RTCS in coleoptilar nodes is confined to emerging shoot-borne root primordia. Sequence analyses of the RTCS and RTCL upstream genomic regions identified auxin response elements. Reverse transcriptase-PCR revealed that both genes are auxin induced. Microsynteny analyses between maize and rice genomes revealed co-linearity of 14 genes in the RTCS region. We conclude from our data that RTCS and RTCL are auxin responsive genes involved in the early events that lead to the initiation and maintenance of seminal and shoot-borne root primordia formation. PMID- 17425723 TI - Effects of immunosuppressive drugs on HIV infection: implications for solid-organ transplantation. AB - With the advent of highly active antiretroviral therapy (HAART), HIV infection has become a chronic disease. Various end-stage organ failures have now become common co-morbidities and are primary causes of mortality in HIV-infected patients. Solid-organ transplantation therefore has been proposed to these patients, as HIV infection is not anymore considered an absolute contraindication. The initial results of organ transplantation in HIV-infected patients are encouraging with no differences in patient and graft survival compared with non-HIV-infected patients. The use of immunosuppressive drug therapy in HIV-infected patients has so far not shown major detrimental effects, and some drugs in combination with HAART have even demonstrated possible beneficial effects for specific HIV settings. Nevertheless, organ transplantation in HIV-infected patients remains a complex intervention, and more studies will be required to clarify open questions such as long-term effects of drug interactions between antiretroviral and immunosuppressive drugs, outcome of recurrent HCV infection in HIV-infected patients, incidence of graft rejection, or long-term graft and patient survival. In this article, we first review the immunological pathogenesis of HIV infection and the rationale for using immunosuppression combined with HAART. We then discuss the most recent results of solid-organ transplantation in HIV-infected patients. PMID- 17425724 TI - Renal dysfunction in liver transplant patients: comparing patients transplanted for liver tumor or acute or chronic disease. AB - Liver transplant patients are susceptible to renal dysfunction through a number of mechanisms. Our aim was to investigate if renal function differs among transplant indication groups. Consecutive liver transplantations (396) were divided in three groups: 277 with chronic liver disease (CLD), 90 with acute liver failure (ALF), and 29 with liver tumor. Data were recorded before and after transplantation. The glomerular filtration rate (GFR) was based on Cockcroft Gault formula and renal function staged using the National Kidney Foundation guidelines. On the transplantation day, 4%, 15%, and 0% of patients in the CLD, ALF, and tumor groups, respectively, showed severely decreased GFR (10(5) IU/mL in four patients (15%) at 20-29 month in monoprophylaxis group and six (18%) at 4-35 months in combination group. Recurrent hepatitis was seen in two patients (8%) at 27 and 45 months and monoprophylaxis group and three (9%) at 21-35 months in combination group. The rate of recurrence was not statistically different between two groups. CONCLUSION: LAM monoprophylaxis seemed to be effective for OLT recipients with HBV infection who had non-replicating HBV at transplantation. HBIG administration may play a less valuable role in preventing HBV recurrence in this group of patients. PMID- 17425741 TI - Significance of microvascular thrombosis in renal allografts: role of ex vivo thrombolytic therapy. AB - BACKGROUND: Microvascular thrombosis is an uncommon pathologic finding in cadaveric kidneys. It is associated with disseminated intravascular coagulation most frequently associated with head injuries and massive blood transfusions. The high rate of non-function published in the literature portends an ominous significance for transplanted organs, which have been discarded by many transplant centers. METHODS: In this report, we document the effectiveness of ex vivo thrombolytic therapy with activase, a plasminogen activator, injected intra arterially in 12 kidneys with extensive histologic intraglomerular microvascular thrombosis prior to transplantation. All cold stored cadaveric kidneys were flushed for 50 min with 50 mg of activase diluted in University of Wisconsin solution. Pre-flush and post-flush wedge biopsies were performed on the allografts. RESULTS: All biopsies showed complete resolution of the intraglomerular microthrombi following activase flush. One kidney clotted in the third week. Three patients developed delayed graft function (25%) of which one experienced allograft thrombosis following one month after transplantation. All remaining grafts are still functioning at a mean follow up of 55.1 months (range: 23-71) with a mean serum creatinine of 1.8 mg/dL (range: 0.8-2.5). CONCLUSIONS: Back table flush with activase rapidly lyses severe intraglomerular microthrombi making them successfully transplantable. Pre-treatment with activase seems to be an effective therapeutic intervention for kidneys with massive intraglomerular thrombosis. PMID- 17425742 TI - Twenty-five consecutive isolated intestinal transplants in adult patients: a five yr clinical experience. AB - PATIENTS AND METHODS: Between December 2000 and December 2005, 25 isolated intestinal transplants from cadaveric donors have been performed for short gut syndrome (short bowel syndrome, 52%), chronic intestinal pseudo-obstruction (24%), Gardner syndrome (16%), radiation enteritis (4%) and massive intestinal angiomatosis (4%). Indications for transplantation were: loss of venous access, recurrent sepsis due to central line infection, major electrolyte and fluid imbalance. Liver dysfunction was present in 13 cases. All patients were adult; median age was 36.3 yr and mean weight at transplantation 61.6 kg. All recipients were on life-threatening parenteral nutrition for a mean time of 23.7 months. Mean donor/recipient body weight ratio was 1.08. Rejection monitoring was accomplished by graft ileoendoscopies and intestinal biopsies through the temporary ileostomy. Our immunosuppressive regimen was based on induction therapy with three different protocols: daclizumab for induction, tacrolimus and steroids as maintenance therapy; alemtuzumab for induction and low-dose tacrolimus as maintenance; thymoglobulin for induction and maintenance based on low-dose tacrolimus. Closure of the abdomen at the end of transplantation represented a technical problem with several options performed: graft reduction, only skin closure, prothesic meshes, abdominal closure in two steps, cutaneous flaps and abdominal wall transplant in one case. RESULTS: The mean hospital stay was 37 days. The mean follow-up 27 months. Twenty patients are alive (80%) with two- and five-yr patient survival rate of 80% and 66%; mortality rate was 20% due to sepsis in all cases. Our two- and five-yr graft survival rate is 76% and 64%, graftectomy rate was 16%. Sixteen grafts are working properly, with no need of parenteral nutrition. We diagnosed 35 mild acute cellular rejection (ACRs), seven moderate ACRs and three severe ACRs (two needed graftectomy). We experienced two episodes of chronic rejection biopsy-proven. Rapamicine was added in case of renal failure or biopsy-proven intestinal rejection. Graft-vs.-host disease was not seen in our series while post-transplant lymphoproliferative disease in two cases. After discharge, the most common indication for medical support was dehydration. The abdominal wall transplant did not experience any rejection. DISCUSSION AND CONCLUSIONS: Induction therapy has reduced the amount of postoperative immunosuppressive agents, especially in the first period, lowering the risk of renal failure and sepsis and the mucosal surveillance protocol for early detection of rejection dramatically reduced the number of severe ACR. PMID- 17425743 TI - Paired kidney donation by shipment of living donor kidneys. AB - It is apparent from calculations that for paired kidney donation programs, a national program will provide optimum benefit. To obviate major problems associated with donors traveling long distances, we propose shipping donor kidneys. Evidence is provided from the United Network for Organ Sharing (UNOS) Kidney Transplant Registry, that 14,873 immediate functioning kidneys from deceased head-trauma donors with an average cold ischemia time of 18.3 h had 85.7% three-yr graft survival compared with 87.8% survival of 23 369 transplants from living donors with 2.4 h of ischemia. Grafts from 10,368 deceased donors with 13-24 h cold ischemia time had three-yr graft survival of 82.6% compared with 84.2% for 1153 transplants with up to six h cold ischemia time. After adjusting for major confounding factors, cold ischemia does not significantly influence graft survival. We conclude that shipment of donor kidneys can be performed safely and will significantly increase paired donor transplants. PMID- 17425744 TI - Renal function with cyclosporine C2 monitoring, enteric-coated mycophenolate sodium and basiliximab: a 12-month randomized trial in renal transplant recipients. AB - BACKGROUND: Cyclosporine exposure, as estimated by the area under the curve (AUC), predicts outcomes in renal transplantation. Cyclosporine concentration at two h post-dose (C(2)) has been shown to be the most reliable, single-point surrogate marker for AUC. The objective of this study was to measure renal function beyond month 2 post-transplant using two different C(2) maintenance targets in combination with enteric-coated mycophenolate sodium (EC-MPS), corticosteroids, and basiliximab induction. METHODS: In this open-label, multicenter trial, renal transplant recipients entered one of two randomized groups at day 61 post-transplant: group A (higher-C(2) range) or group B (lower C(2) range). RESULTS: Patients (164) were recruited, and 141 patients were entered the randomized groups (group A, n = 66; group B, n = 75). At 12 months, the mean calculated creatinine clearance was significantly greater in group B than in group A (79.2 vs. 71.0 mL/min, p < 0.05). Biopsy-proven acute rejection occurred in 14.7% patients in group B and in 24.2% patients in group A (n.s.). During the 12-month trial, 17.7% patients discontinued EC-MPS because of adverse events. Group B (44.0%) had fewer serious adverse events when compared with group A (62.1%; p = 0.04). Overall patient and graft survival were 99.4% and 95.7% respectively. Among 99 high-risk patients (i.e., African-American race, previous transplant, PRA >35% or >4 HLA mismatches), mean creatinine clearance at 12 months was 65.6 mL/min and biopsy-proven rejection occurred in 20.2% patients. CONCLUSIONS: Low cyclosporine C(2) levels are associated with improved renal function compared with higher C(2) levels when used in conjunction with EC-MPS, steroids and basiliximab induction. EC-MPS with low cyclosporine C(2) levels, corticosteroids and basiliximab provides excellent renal function with good efficacy even in high-risk patients. PMID- 17425745 TI - The fate of recipient-derived hepatocytes in sex-mismatched liver allograft following liver transplantation. AB - BACKGROUND: ''Bone marrow-derived stem cells'' have attracted great attention as potential candidates for liver-directed gene therapy and as a tool for regenerative medicine. However, the fate of these cells is not well-known. The aim of this present study was to investigate the fate of ''recipient-derived repopulated hepatocytes'' in sex-mismatched liver allografts in individuals following liver transplantation during systematic longitudinally performed liver biopsies. METHODS: Paraffin-embedded sex-mismatched liver biopsy samples of nine recipients (male/female ratio 5/4; mean age: 39.7 yr) were reviewed. Double labeling with immunohistochemistry for hepatocytes and recipient-specific bone marrow-derived cells and fluorescence in-situ hybridization for visualizing X and Y chromosomes were performed. These slides were examined systematically using an image analyzer system (Olympus microscope; Cyto-Vision, Applied Imaging, Biosciences Centre, Newcastle, UK). Only cells with two nuclear spots were considered for interpretation. RESULTS: The mean times from transplantation to first biopsy and between the first and the second biopsies were 5.9 and 20.9 months respectively. The proportion of recipient-derived repopulated hepatocytes was significantly decreased in the late biopsies when compared with the early biopsies (p = 0.001). All nine samples of the first biopsies had demonstrated recipient-derived hepatocyte repopulation, with a mean of 2.0%, whereas only seven of nine samples of the second biopsies had demonstrated recipient-derived hepatocyte repopulation with a low mean of 0.5% (p = 0.001). CONCLUSION: Based on these results, we suggest that ''recipient-specific bone marrow-derived hepatocyte repopulation'' in liver allograft during tissue injury is a relatively early event. PMID- 17425746 TI - Glutathione S-transferase A1 polymorphisms and acute graft-vs.-host disease in HLA-matched sibling allogeneic hematopoietic stem cell transplantation. AB - Busulfan and the metabolites of cyclophosphamide are conjugated with glutathione and catabolized by enzymes of the cytosolic glutathione S-transferases family. There are clearly linked single nucleotide polymorphisms in the promoter region of the glutathione S-transferase A1 gene (i.e., GSTA1*A, -567T, -69C and -52G; GSTA1*B, -567G, -69T and -52A). We assessed whether the clinical outcomes, including acute graft-vs.-host disease, of 61 patients with hematological malignancies, following HLA-matched sibling allogeneic stem cell transplantation using busulfan/cyclophosphamide conditioning are altered by glutathione S transferase A1 genotypes. Globally, grade II-IV acute graft-vs.-host disease developed in 13 patients (21%). Grade II-IV acute graft-vs.-host disease developed in 15.2% of 46 patients with GSTA1*A/*A diplotype and in 40.0% of 15 patients with GSTA1*A/*B or GSTA1*B/*B diplotype (p = 0.04). Moreover, this relationship between GSTA1*A/*A diplotypes and lower incidence of acute graft-vs. host disease was independent of the age, gender, stem cell source, and disease status. The incidences of acute skin graft-vs.-host disease were 7% (3/46) in patients with GSTA1*A/*A and 27% (4/15) in patients without GSTA1*A/*A (p = 0.009, univariate; p = 0.01, multivariate). Acute hepatic graft-vs.-host disease developed in 6 (13%) of 46 patients with the GSTA1*A/*A diplotype and in 4 (27%) of 15 patients without this diplotype (p = 0.09, univariate; p = 0.12, multivariate). Ten patients (16%) developed hepatic veno-occlusive disease. No significant difference was found in the incidence of hepatic veno-occlusive disease between patients with and without the GSTA1*A/*A diplotype (19.6% vs. 6.7%; p = 0.24). We conclude that the GSTA1*A/*A diplotype is an independent protective factor against acute graft-vs.-host disease, especially for skin graft vs.-host disease, and probably for hepatic graft-vs.-host disease, in patients using busulfan/cyclophosphamide conditioning. The identification of glutathione S transferase A1 genotypes prior to allogeneic stem cell transplantation could allow conditioning regimens and graft-vs.-host disease prophylaxis to be modified to improve outcome. PMID- 17425747 TI - Histologic eosinophilia as an aid to diagnose acute cellular rejection after living donor liver transplantation. AB - The significance of histologic eosinophilia in the diagnosis of acute cellular rejection (ACR) after living donor liver transplantation was evaluated. A retrospective analysis was performed on 185 liver biopsy specimens to determine the presence of eosinophil infiltration around the portal tracts. Data were collected and analyzed to determine whether there was a correlation between ACR and the maximum eosinophil counts per portal triad (Em) and the rate of portal triads that included at least one eosinophil (Er). A receiver operating characteristic curve revealed the best cut-off value of Em and Er as 2% and 8% respectively. The sensitivity and specificity of an Em of two to predict ACR were 54% and 84% respectively. The sensitivity and specificity of Er were 72% and 65% respectively. One-way analysis of variance revealed that both Em and Er correlated with ACR severity. Histologic eosinophilia can be a useful parameter for confirming the occurrence of ACR and for evaluating ACR severity. PMID- 17425748 TI - Examining the association between media coverage of organ donation and organ transplantation rates. AB - Research addressing the organ shortage in the USA has examined multiple factors influencing one's decision to become an organ donor. One of these research lines addresses media coverage of organ donation. The present investigation seeks to advance this research line by examining the association between organ donation media coverage and organ transplantation rates. A content analysis spanning January 1990 to December 2005 of three television networks reveals an overall positive association between coverage and transplantation rates. The implications of our findings are discussed along with recommendations for practitioners and advocates alike. PMID- 17425749 TI - Disability following kidney transplantation: the link to medication coverage. AB - There is no uniformity regarding patient disability following kidney transplantation. Given improved results of patient and graft survival, and the link between insurance, medication coverage and disability, efforts must be made to define disability after a successful transplant. We conducted an individual questioner study of kidney transplant patients to determine factors relating to patient-perceived disability. Seventy patients participated in the study. Patient perception of disability did not correlate with education or ethnicity. Most patients believed they were disabled on dialysis and this did not change following transplantation. While 42 (60%) of the patients felt that they could work, either full-time or part-time, only 20 (28%) were actually working or in school. Most patients believe that working will eliminate disability status and, therefore, insurance and medication coverage. Patients considered disability more related to their status as a kidney transplant patient than any specific physical limitations. The link, whether real or perceived, between 'disability' and immunosuppressive medication coverage is a significant barrier for many patients. The transplant community must reach some degree of consensus regarding post transplant activity restrictions. The transplant community needs to find a way to take an active role in post-transplant education and employment. PMID- 17425750 TI - Liver transplantation without isoniazid prophylaxis for recipients with a history of tuberculosis. AB - Tuberculosis remains one of the most serious infections after organ transplantation. Isoniazid prophylaxis for liver transplant recipients with a history of tuberculosis is generally recommended. However, its benefit is controversial because of potential hepatotoxicity of isoniazid. It is crucial to determine appropriate post-transplant managements for the recipients with a history of tuberculosis. The purpose of this study was to investigate the necessity of isoniazid prophylaxis for liver transplant recipients who had a history of tuberculosis. The medical records of 1116 liver transplant recipients were studied, of whom seven had a history of tuberculosis (0.63%). One who underwent living-donor liver transplantation for fulminant hepatic failure was excluded from evaluation because of early death, caused by bacterial sepsis two months after transplantation, although reactivation of tuberculosis was not observed. The median observation period after transplantation was 25.5 months (range 12-82). Reactivation of tuberculosis did not occur in any of these six patients. In conclusion, we could not find rationale for isoniazid prophylaxis in liver transplant recipients with past diagnosis of tuberculosis, when the disease is considered to be inactive. Tuberculosis should be considered as cause of post transplant infections, and careful post-transplant observations are essential for an early diagnosis. PMID- 17425751 TI - Does mini-incision donor nephrectomy improve quality of life in living kidney donors? AB - BACKGROUND: Living kidney donation helps to avoid or reduce the time period of dialysis and on waiting lists in patients requiring a new organ. Mini-incision donor nephrectomy (MIDN) shows to result in better clinical outcome in comparison with traditional open donor nephrectomy (ODN). This study was performed to evaluate the impact of different surgical procedures on the quality of life (QoL) in patients that underwent donor nephrectomy. METHODS: The aim of the study was to detect differences in QoL assessed with the Short Form-36 Version 2 (SF-36v2) questionnaire between MIDN (n = 34) and ODN (n = 36). Furthermore, the development of QoL from prior to surgery until one yr afterwards, as well as outcomes of QoL in comparison with norm-based scores was investigated. RESULTS: Sixty-one of 70 patients, which is 87% (MIDN: 86%, ODN: 88%) resent a whole set questionnaires. QoL was similar at all time-points (prior to surgery, one wk, three months and one yr) in both groups. A tendency of better QoL in MIDN (Bodily Pain) after one wk was detectable (p = 0.075). Physical Component Summaries (PCS) significantly decreased from prior to surgery until one wk after surgery (p = 0.001) and improved significantly until three months (MIDN: p = 0.006, ODN: p = 0.001) and also until one yr after surgery (p = 0.002). Mental Component Summaries (MCS) were stable throughout the whole investigated time period. In comparison with norm-based scores, MIDN (p = 0.005) and ODN (p = 0.001) showed significantly higher PCS prior to, lower scores one wk after (p = 0.001), similar scores three months after and better scores (MIDN: p = 0.023, ODN: 0.015) one yr after surgery. Mental Component Scores were similar in both prior to and one wk after surgery. After three months and one yr scores were significantly better in MIDN (three months: p = 0.049, one yr: p = 0.037) and ODN (three months: 0.020, one yr: 0.073). CONCLUSION: Quality of life after living donor nephrectomy is not influenced by the surgical technique. Nevertheless the standardized instrument of the SF-36v2 Health Survey is a useful, practicable and universally interpretable tool to gain and estimate recovery from surgical procedures in the perioperative period and its development thereafter. PMID- 17425752 TI - Thrombotic microangiopathy after simultaneous pancreas-kidney transplantation. AB - Thrombotic microangiopathy (TMA) is rare after transplantation and is associated with a high incidence of kidney graft dysfunction. Between December 2000 and March 2006, 136 simultaneous pancreas-kidney transplantations were performed with an incidence of TMA of 5.1% (71.4% localized to kidney allograft). All cases were diagnosed during the first three months and were attributed to tacrolimus; 74% were women. Systemic TMA presented higher values of lactate dehydrogenase (2658 +/- 659 U/L vs. 1331 +/- 473 U/L, p = 0.04) and a greater decrease in hematocrit (45.8 +/- 17.7% vs. 19.2 +/- 6%, p = 0.02) than in localized TMA. Acute kidney rejection complicated almost 90% of the cases with 43% of kidney graft lost. Tacrolimus was switched to sirolimus and fresh-frozen plasma was administered. Creatinine clearance after a mean follow-up of two yr was 100.7 mL/min/1.73 m(2) and 57.9 mL/min/1.73 m(2) in patients with systemic and localized TMA, respectively. In conclusion, sirolimus is an alternative to TMA associated with tacrolimus. PMID- 17425753 TI - Nateglinide improves postprandial hyperglycemia and insulin secretion in renal transplant recipients. AB - BACKGROUND: Postprandial hyperglycemia (PPHG) frequently occurs among renal transplant recipients (RTR). Reduced early insulin response (EIR) after a meal leads to impaired suppression of endogenous glucose production and subsequently PPHG, which is a risk factor for cardiovascular disease. Nateglinide is a rapid acting insulin secretagogue inducing an EIR after a meal. Our main objective was to investigate the safety and effect of nateglinide treatment on postprandial plasma glucose excursions and insulin secretion in RTR with PPHG. PATIENTS AND METHODS: A total of 14 Caucasian RTR with new-onset diabetes mellitus (NODM; n = 6) or impaired glucose tolerance (IGT; n = 8) were included. The insulin response and glucose excursions were measured for 240 min after a standardized liquid meal at baseline and after two-wk treatment with nateglinide. RESULTS: Treatment with nateglinide was followed by a significant decrease in mean two-h plasma glucose from 10.5 mmol/L (3.1) to 7.6 mmol/L (2.1; p < 0.001) and a decline in total postprandial area under the curve (AUC) of glucose concentration (p < 0.001). Both estimated EIR and the late insulin response increased significantly (p = 0.008 and p = 0.003, respectively). No serious adverse event was observed during the study period. CONCLUSIONS: Treating RTR with nateglinide for two-wk significantly improved PPHG, increased the insulin response following a standardized meal and was well tolerated. PMID- 17425754 TI - Multi-drug resistance gene-1 (MDR-1) haplotypes and the CYP3A5*1 genotype have no influence on ciclosporin dose requirements as assessed by C0 or C2 measurements. AB - The intestinal efflux pump P-glycoprotein (P-gp), the product of the multi-drug resistance-1 (MDR-1) gene, significantly influences the pharmacokinetics of several drugs. Ciclosporin is a substrate for P-gp and is metabolized by cytochrome P450 (CYP) 3A enzymes. P-gp activity is affected by several known single nucleotide polymorphisms (SNPs) and haplotypes. MDR-1 genotypes of SNPs C1236T, G2677T/A and C3435T, as well as haplotypes C-G-C and T-T-T and CYP3A5*1 genotype (predictive of CYP3A5 expression), were related to ciclosporin blood concentrations measured at both 0 and 2 h after drug dosing in 197 stable renal transplant patients. Significant differences (of a magnitude unlikely to be relevant clinically) in dose-normalized blood ciclosporin concentrations were found only between MDR-1 genotypes of the C1236T SNP and between haplotype groups C-G-C and T-T-T in patients that were expressers of CYP3A5. MDR-1 SNPs and haplotypes and also CYP3A5*1 genotype, do not appear to have a major influence on ciclosporin pharmacokinetics. PMID- 17425755 TI - Longer survival of liver transplant recipients with hepatitis virus coinfections. AB - Hepatitis virus coinfections [HBV plus HCV coinfection (HBV/HCV) or HBV plus HDV coinfection (HBV/HDV)] may progress more rapidly to cirrhosis than hepatitis B or C monoinfections in immunocompetent patients. Only limited information is available on the outcome of coinfected patients after liver transplantation. We studied survival rates of 204 patients with viral hepatitis transplanted at our center between 1972 and 1997. HBV/HDV and HBV/HCV coinfections were present in 23 and nine individuals, respectively, while 97 patients had monoinfection by HCV and 75 had HBV monoinfection. Survival of coinfected patients was significantly longer than that of monoinfected patients (14.4 +/- 0.9 vs. 8.5 +/- 0.6 yr; p = 0.0003). The same was true for graft survival (p = 0.0002). In Cox's regression, viral coinfection (p = 0.0001), absence of hepatocellular carcinoma (HCC) (p = 0.00001) and no retransplantation (p = 0.02) were independently associated with patient survival. After exclusion of patients with HCC (n = 62), survival of coinfected patients was still significantly longer than that of monoinfected individuals (p = 0.002). The improved outcome was similar for both HBV/HDV and HBV/HCV coinfections. In contrast to immunocompetent patients, individuals with multiple hepatitis virus infections had an improved outcome after liver transplantation. Thus, viral coinfections may be associated with ameliorated courses of diseases under certain conditions. PMID- 17425756 TI - Steroids in intestinal transplantation. AB - BACKGROUND: Recently, new immunosuppressive protocols after intestinal transplantation have been proposed to avoid steroids use and their adverse effects. We evaluated the impact of steroids on survival and post-transplant complications in our experience. PATIENT AND METHODS: In our retrospective study we considered the mean daily dosage of steroids received by 25 patients after intestinal/multivisceral transplantation (minimal follow-up was six months). We analyzed graft and patient survival rates, correlation with rejection and infectious episodes and steroids side effects. RESULTS: After a mean follow-up of three yr, we did not find any significant difference in steroid doses between our immunosuppressive protocols. Patients with a mean dosage of prednisone higher than 20 mg/d experienced a lower graft (p = 0.009) and patient (p = 0.02) survival rate. The side effects of steroids after transplant were similar. Infections were more frequent during steroids administration (p = 0.04). DISCUSSION AND CONCLUSION: Steroids therapy may be useful to treat acute rejection, but in our experience high steroids regimen did not improve graft and patient survival, increasing infectious rate. We assumed that high dose of steroids can be avoided as maintenance therapy, except in selected cases. PMID- 17425757 TI - Mini-incisions by lombotomy or subcostal access in living kidney donors: a randomized trial comparing pain, safety, and quality of life. AB - OBJECTIVES: The aim of this study was to compare two mini-incision techniques and judge the impact on the quality of life, pain, and safety of living kidney donors. PATIENTS AND METHODS: From March through September 2003, a prospective randomized study with 60 donors had nephrectomy performed - 30 through a lombotomy and another 30 patients underwent subcostal mini-incisions. The same anesthetic procedure was used for both groups. All patients were evaluated from baseline (T0) to day 90 after surgery. Pain evaluation included visual analog scale (VAS) and drug usage. To assess quality of life (QOL), the questionnaire SF 36 was used and surgical outcomes were also checked. RESULTS: Sixty patients (41.6 +/- 8.9 yr old) were included in the protocol. Regarding incision length and blood loss, no statistical difference was observed. However, irrespective to the site of the mini-incision, patients with body mass index (BMI) higher than 25 kg/m(2) had significantly longer incision length as well as higher blood loss. There were no complications. No significant difference in tramadol or in pain perception was observed between groups. QOL was also not different between groups, however, there was a significant loss with subsequent return to baseline levels. CONCLUSION: The position of the mini-incision (lombotomy or subcostal) has no significant impact on surgical outcomes, pain perception, and QOL of living kidney donors. Mini-incision techniques represent fast and safe approaches to perform nephrectomy in the healthy population. Special care must be taken in obese patients in order to minimize surgical complications. PMID- 17425758 TI - Enalapril/amlodipine combination in cyclosporine-treated renal transplant recipients: a prospective randomized trial. AB - BACKGROUND: Most hypertensive renal transplant recipients require two or more antihypertensive medications to achieve blood pressure control. However, which medications must be combined is still a matter of debate. METHODS: A prospective randomized open-label blinded evaluation trial comparing the six-month effects of the amlodipine-enalapril combination (n = 32) vs. enalapril alone (n = 33) and vs. amlodipine alone (n = 34) on arterial pressure, renal function, albuminuria and tolerability. RESULTS: At six months, diastolic arterial pressure was more adequately controlled (i.e., <90 mmHg) in the combination group than in the amlodipine and enalapril groups (100% vs. 82.4% and 84.8%, respectively, p = 0.038). The same trend was observed for systolic arterial pressure (65.6% vs. 58.8% and 51.5%, NS). The six-month change in albuminuria was similar in the combination group and in the enalapril group (-64.7% vs. -59.5%); however, patients in the combination group exhibited a greater reduction in albuminuria than in the amlodipine group (-64.7% vs. -29.0%, p = 0.002). As compared with baseline values, serum creatinine and potassium remained unchanged in the combination group, whereas they increased by 9 +/- 12 micromol/L (p = 0.01) and by 0.2 +/- 0.4 mmol/L (p < 0.01), respectively, in the enalapril group. The cyclosporine trough levels remained unchanged in the combination group, but increased in the amlodipine group. CONCLUSION: Angiotensin-converting enzyme inhibitor (ACEI)-calcium-channel blocker (CCB) combination controls arterial pressure more adequately than ACEI alone or CCB alone, reduces albuminuria and may prevent the ACEI-induced initial rise in serum creatinine. PMID- 17425759 TI - Influence of economic and demographic factors on quality of life in renal transplant recipients. AB - BACKGROUND: The purpose of this study was to determine the influence of annual income, Medicare status, and demographic variables on the health-related quality of life (HQoL) of renal transplant recipients. METHODS: A cross-sectional survey was mailed to 146 Georgia renal transplant recipients who had functional grafts. Data were collected using the SF-12 Health Survey (version 2), a demographics survey, and 2003 tax documents. One-way ANOVAs and Pearson's R correlations were used to examine relationships between annual income, Medicare status, demographic variables and SF-12 scores. Significant variables were included in stepwise multiple regression analyses. RESULTS: Data from 130 participants (89% response rate) were collected. Recipients with no Medicare coverage had significantly higher scores on the Physical Functioning and Role Physical SF-12 scales (p = 0.005) compared to recipients with Medicare. Annual income was positively correlated with General Health (p < 0.05). Age and race were significant predictors of Vitality (p = 0.004) and Physical Component Summary (p < 0.001) scores. Age, race, and Medicare status were significant predictors of Physical Functioning and Role Physical scores (p < 0.001). Age, annual income, race, and years post-transplant were significant predictors of General Health score (p < 0.001). Age was the sole predictor of Bodily Pain score (p = 0.002), and marital status was the sole predictor of Social Functioning score (p = 0.005). CONCLUSIONS: Interventions designed to offset financial barriers may be needed to bolster renal transplant recipients' HQoL. PMID- 17425761 TI - Know your colour vision challenges. PMID- 17425762 TI - Assessment of inherited colour vision defects in clinical practice. AB - BACKGROUND: Colour vision deficiency (CVD) has a high prevalence and is often a handicap in everyday life. Those who have CVD will be better able to adapt and make more informed career choices, if they know about their deficiency. The fact that from 20 to 30 per cent of adults with abnormal colour vision do not know they have CVD suggests that colour vision is not tested as often as it should be. This may be because of practitioner uncertainty about which tests to use, how to interpret them and the advice that should be given to patients on the basis of the results. The purpose of this paper is to recommend tests for primary care assessment of colour vision and provide guidance on the advice that can be given to patients with CVD. METHODS: The literature on colour vision tests and the relationship between the results of the tests and performance at practical colour tasks was reviewed. RESULTS: The colour vision tests that are most suitable for primary care clinical practice are the Ishihara test, the Richmond HRR 4th edition 2002 test, the Medmont C-100 test and the Farnsworth D15 test. These tests are quick to administer, give clear results and are easy to interpret. Tables are provided summarising how these tests should be interpreted, the advice that can be given to CVD patients on basis of the test results, and the occupations in which CVD is a handicap. CONCLUSION: Optometrists should test the colour vision of all new patients with the Ishihara and Richmond HRR (2002) tests. Those shown to have CVD should be assessed with the Medmont C-100 test and the Farnsworth D15 test and given appropriate advice based on the test results. PMID- 17425763 TI - The effect of vertical gaze position on ocular tear film stability. AB - BACKGROUND: An enlarged exposed ocular surface area is known to degrade tear film stability. Little is known of how tear film stability alters with vertical gaze and the change of palpebral fissure height. METHODS: Tear film break-up time (TBUT; fluorescent dye) and palpebral fissure height were measured in three vertical gaze positions (primary position and 20 degrees up and 20 degrees down) on 23 healthy volunteers. The effect of gaze direction on TBUT was analysed by ANOVA. RESULTS: Gaze had a significant influence on tear film stability [F(2,44)=27.6, p<0.001]. In up-gaze with an enlarged palpebral fissure height (11.1 +/- 1.4 mm), TBUT was degraded (17.3 +/- 12.7 seconds). In down-gaze and decreased palpebral fissure height (6.7 +/- 1.2 mm) TBUT was extended (43.5 +/- 35.4 seconds). CONCLUSION: Ocular tear film stability alters with vertical gaze. Our hypothesis is that the change of the palpebral fissure height and the exposed ocular surface area alters the thickness of the lipid layer and with it tear film evaporation. PMID- 17425764 TI - Microbial keratitis in orthokeratology: the Australian experience. AB - BACKGROUND: This study was conducted to investigate the demographics of orthokeratology (OK) practice in Australia, to uncover any previously undocumented cases of serious adverse responses in OK, including microbial keratitis (MK), and to review the demographics of MK in OK in Australia. METHODS: A questionnaire was sent to the 62 members of the Orthokeratology Society of Australia (OSA). Questions related to aspects of their OK practice, demographics of their OK patient base and any adverse responses to OK lens wear that they had encountered. RESULTS: Thirty-three questionnaires (53 per cent) were returned. OSA members have been fitting OK lenses for a median of 7.5 years. OK patients were predominantly female, Caucasian, aged between 15 and 39 years and wearing lenses in an overnight modality. In addition to two cases reported previously, the survey uncovered seven further cases of MK in OK patients over an eight-year period. The infecting organism was Pseudomonas aeruginosa in four cases, Acanthamoeba spp. in two cases and unknown in three cases. There was no loss of visual acuity in seven cases. One case resulted in vision of counting fingers at one metre and another case resolved with 6/12 visual acuity. Non-compliance with instructions on lens care and after-care was reported in seven of nine cases of MK. CONCLUSION: Overall, OSA members who responded to the survey have many years of experience in OK. The typical Australian OK patient is in young adulthood, female and Caucasian. A total of nine cases of presumed MK associated with OK have been reported in Australia over an eight-year period and seven of these were new cases uncovered by this survey. Our analysis suggests that the demographics of MK cases in OK reflect the demographics of the OK lens-wearing population. PMID- 17425765 TI - The effect of a compliance enhancement strategy (self-review) on the level of lens care compliance and contamination of contact lenses and lens care accessories. AB - PURPOSE: The aims were to determine the level of compliance of contact lens wearers, to identify which procedures have highest levels of non-compliance and to study the effectiveness of a compliance enhancement strategy on the level of compliance. METHODS: The level of compliance for each subject was evaluated with a self-administered questionnaire together with microbiological examination of their contact lenses, lens cases and lens care solutions. The effect of an intervention to improve the level of compliance was determined by a controlled trial. A regular self-review exercise on proper lens handling was given to the test group once every three months for 12 months. The levels of compliance and contamination of contact lenses and lens care accessories between test and control groups were compared at the end of the 12-month period to evaluate the effect of the intervention. RESULTS: Sixty-five subjects who were experienced contact lens wearers were recruited and 60 completed the study. All showed some degree of non-compliance in the care of their contact lenses and lens accessories. Most (about 60 per cent) were non-compliant with at least six of a total of 15 lens care procedures. The most common non-compliant behaviour among contact lens wearers was associated with the care of the lens case. By the end of the study period, our compliance enhancement strategy did not appear to have had a significant effect on the behaviour of our subjects, except for improvement in the care of lens cases. Compliance for other procedures improved in both test and control groups. CONCLUSION: All subjects showed some degree of non-compliance and the spectrum of non-compliance was wide. The poorest levels of compliance were associated with care of the lens case, which was also the most frequently contaminated item. Our compliance enhancement strategy showed a significant difference in improvement in compliance between the intervention and control groups only in the care of lens cases. PMID- 17425766 TI - The effect of font size and type on reading performance with Arabic words in normally sighted and simulated cataract subjects. AB - BACKGROUND: Previous investigations have shown that reading is the most common functional problem reported by patients at a low vision practice. While there have been studies investigating effect of fonts in normal and low vision patients in English, no study has been carried out in Arabic. Additionally, there has been no investigation into the use of optimum print sizes or fonts that should be used in Arabic books and leaflets for low vision patients. METHODS: Arabic sentences were read by 100 normally sighted volunteers with and without simulated cataract. Subjects read two font types (Times New Roman and Courier) in three different sizes (N8, N10 and N12). The subjects were asked to read the sentences aloud. The reading speed was calculated as number of words read divided by the time taken, while reading rate was calculated as the number of words read correctly divided by the time taken. RESULTS: There was an improvement in reading performance of normally sighted and simulated visually impaired subjects when the print size increased. There was no significant difference in reading performance between the two types of font used at small print size, however the reading rate improved as print size increased with Times New Roman. CONCLUSION: The results suggest that the use of N12 print in Times New Roman enhanced reading performance in normally sighted and simulated cataract subjects. PMID- 17425767 TI - Bilateral intracavernous carotid artery aneurysms presenting as progressive cranial nerve palsies. AB - Bilateral intracavernous carotid artery aneurysms are rare and can present with different clinical findings. We report such a case presenting with progressive cranial nerve palsies due to gradual increase in the size of aneurysms. This case demonstrates the clinician's difficulty in diagnosis when isolated cranial nerve palsy is the sole presenting sign. PMID- 17425768 TI - An unusual case of a Forster-Fuchs' spot. AB - Myopia has many pathological features in the eye. In this report, we review the unusual case of a young gentleman who developed a Forster-Fuchs' spot at his macula, with his fundus and fluorescein angiographic findings, and current prognosis and management. PMID- 17425774 TI - Web-based tools can be used reliably to detect patients with major depressive disorder and subsyndromal depressive symptoms. AB - BACKGROUND: Although depression has been regarded as a major public health problem, many individuals with depression still remain undetected or untreated. Despite the potential for Internet-based tools to greatly improve the success rate of screening for depression, their reliability and validity has not been well studied. Therefore the aim of this study was to evaluate the test-retest reliability and criterion validity of a Web-based system, the Internet-based Self assessment Program for Depression (ISP-D). METHODS: The ISP-D to screen for major depressive disorder (MDD), minor depressive disorder (MinD), and subsyndromal depressive symptoms (SSD) was developed in traditional Chinese. Volunteers, 18 years and older, were recruited via the Internet and then assessed twice on the online ISP-D system to investigate the test-retest reliability of the test. They were subsequently prompted to schedule face-to-face interviews. The interviews were performed by the research psychiatrists using the Mini-International Neuropsychiatric Interview and the diagnoses made according to DSM-IV diagnostic criteria were used for the statistics of criterion validity. Kappa (kappa) values were calculated to assess test-retest reliability. RESULTS: A total of 579 volunteer subjects were administered the test. Most of the subjects were young (mean age: 26.2 +/- 6.6 years), female (77.7%), single (81.6%), and well educated (61.9% college or higher). The distributions of MDD, MinD, SSD and no depression specified were 30.9%, 7.4%, 15.2%, and 46.5%, respectively. The mean time to complete the ISP-D was 8.89 +/- 6.77 min. One hundred and eighty-four of the respondents completed the retest (response rate: 31.8%). Our analysis revealed that the 2-week test-retest reliability for ISP-D was excellent (weighted kappa = 0.801). Fifty-five participants completed the face-to-face interview for the validity study. The sensitivity, specificity, positive, and negative predictive values for major depressive disorder were 81.8% and 72.7%, 66.7%, and 85.7% respectively. The overall accuracy was 76.4%. CONCLUSION: The evidence indicates the ISP-D is a reliable and valid online tool for assessing depression. Further studies should test the ISP-D in clinical settings to increase its applications in clinical environments with different populations and in a larger sample size. PMID- 17425775 TI - Choice of treatment for fever at household level in Malawi: examining spatial patterns. AB - BACKGROUND: Although malaria imposes an enormous burden on Malawi, it remains a controllable disease. The key strategies for control are based on early diagnosis and prompt treatment with effective antimalarials. Its success, however, depends on understanding the factors influencing health care decision making at household level, which has implications for implementing policies aimed at promoting health care practices and utilization. METHODS: An analysis of patterns of treatment seeking behaviour among care-givers of children of malarial fever in Malawi, based on the 2000 Malawi demographic and health survey, is presented. The choice of treatment provider (home, shop, or formal hospital care, others) was considered as a multi-categorical response, and a multinomial logistic regression model was used to investigate determinants of choosing any particular provider. The model incorporated random effects, at subdistrict level, to measure the influence of geographical location on the choice of any treatment provider. Inference was Bayesian and based on Markov chain Monte Carlo techniques. RESULTS AND CONCLUSION: Spatial variation was found in the choice of a provider and determinants of choice of any provider differed. Important risk factors included place of residence, access to media, care-giver's age and care factors including unavailability and inaccessibility of care. A greater effort is needed to improve the quality of malaria home treatment or expand health facility utilization, at all levels of administration if reducing malaria is to be realised in Malawi. Health promotion and education interventions should stress promptness of health facility visits, improved access to appropriate drugs, and accurate dosing for home-based treatments. PMID- 17425776 TI - DNA repair gene XRCC1 polymorphisms and bladder cancer risk. AB - BACKGROUND: Cigarette smoking and chemical occupational exposure are the main known risk factors for bladder transitional cell carcinoma (TCC). Oxidative DNA damage induced by carcinogens present in these exposures requires accurate base excision repair (BER). The XRCC1 protein plays a crucial role in BER by acting as a scaffold for other BER enzymes. Variants in the XRCC1 gene might alter protein structure or function or create alternatively spliced proteins which may influence BER efficiency and hence affect individual susceptibility to bladder cancer. Recent epidemiological studies have shown inconsistent associations between these polymorphisms and bladder cancer. To clarify the situation, we conducted a comprehensive analysis of 14 XRCC1 polymorphisms in a case-control study involving more than 1100 subjects. RESULTS: We found no evidence of an association between any of the 14 XRCC1 polymorphisms and bladder cancer risk. However, we found carriage of the variant Arg280His allele to be marginally associated with increased bladder cancer risk compared to the wild-type genotype (adjusted odds ratio [95% confidence interval], 1.50 [0.98-2.28], p = 0.06). The association was stronger for current smokers such that individuals carrying the variant 280His allele had a two to three-fold increased risk of bladder cancer compared to those carrying the wildtype genotype (p = 0.09). However, the evidence for gene-environment interaction was not statistically significant (p = 0.45). CONCLUSION: We provide no evidence of an association between polymorphisms in XRCC1 and bladder cancer risk, although our study had only limited power to detect the association for low frequency variants, such as Arg280His. PMID- 17425777 TI - Factors associated with excessive bleeding in cardiopulmonary bypass patients: a nested case-control study. AB - INTRODUCTION: Excessive bleeding (EB) after cardiopulmonary bypass (CPB) may lead to increased mortality, morbidity, transfusion requirements and re-intervention. Less than 50% of patients undergoing re-intervention exhibit surgical sources of bleeding. We studied clinical and genetic factors associated with EB. METHODS: We performed a nested case-control study of 26 patients who did not receive antifibrinolytic prophylaxis. Variables were collected preoperatively, at intensive care unit (ICU) admission, at 4 and 24 hours post-CPB. EB was defined as 24-hour blood loss of > 1 l post-CPB. Associations of EB with genetic, demographic, and clinical factors were analyzed, using SPSS-12.2 for statistical purposes. RESULTS: EB incidence was 50%, associated with body mass index (BMI) < 26.4 (25-28) Kg/m2, (P = 0.03), lower preoperative levels of plasminogen activator inhibitor-1 (PAI-1) (P = 0.01), lower body temperature during CPB (P = 0.037) and at ICU admission (P = 0.029), and internal mammary artery graft (P = 0.03) in bypass surgery. We found a significant association between EB and 5G homozygotes for PAI-1, after adjusting for BMI (F = 6.07; P = 0.02) and temperature during CPB (F = 8.84; P = 0.007). EB patients showed higher consumption of complement, coagulation, fibrinolysis and hemoderivatives, with significantly lower leptin levels at all postoperative time points (P = 0.01, P < 0.01 and P < 0.01). CONCLUSION: Excessive postoperative bleeding in CPB patients was associated with demographics, particularly less pronounced BMI, and surgical factors together with serine protease activation. PMID- 17425778 TI - Factors associated with dental attendance among adolescents in Santiago, Chile. AB - BACKGROUND: Dental treatment needs are commonly unmet among adolescents. It is therefore important to clarify the determinants of poor utilization of dental services among adolescents. METHODS: A total of 9,203 Chilean students aged 12-21 years provided information on dental visits, oral health related behavior, perceived oral health status, and socio-demographic determinants. School headmasters provided information on monthly tuition and annual fees. Based on the answers provided, three outcome variables were generated to reflect whether the respondent had visited the dentist during the past year or not; whether the last dental visit was due to symptoms; and whether the responded had ever been to a dentist. Aged adjusted multivariable logistic regression models were used to assess the influence of the covariates gender; oral health related behaviors (self-reported tooth brushing frequency & smoking habits); and measures of social position (annual education expenses; paternal income; and achieved parental education) on each outcome. RESULTS: Analyses showed that students who had not attended a dentist within the past year were more likely to be male (OR = 1.3); to report infrequent tooth brushing (OR = 1.3); to have a father without income (OR = 1.8); a mother with only primary school education (OR = 1.5); and were also more likely to report a poor oral health status (OR = 2.0), just as they were more likely to attend schools with lower tuition and fees (OR = 1.4). Students who consulted a dentist because of symptoms were more likely to have a father without income (OR = 1.4); to attend schools with low economic entry barriers (OR = 1.4); and they were more likely to report a poor oral health status (OR = 2.9). Students who had never visited a dentist were more likely to report infrequent tooth brushing (OR = 1.9) and to have lower socioeconomic positions independently of the indicator used. CONCLUSION: The results demonstrate that socioeconomic and behavioral factors are independently associated with the frequency of and reasons for dental visits in this adolescent population and that self-perceived poor oral health status is strongly associated with infrequent dental visits and symptoms. PMID- 17425779 TI - The current MLVA typing scheme for Enterococcus faecium is less discriminatory than MLST and PFGE for epidemic-virulent, hospital-adapted clonal types. AB - BACKGROUND: MLVA (multiple-locus variable-number tandem repeat analysis) is a reliable typing technique introduced recently to differentiate also isolates of Enterococcus faecium. We used the established VNTR (variable number of tandem repeats) scheme to test its suitability to differentiate 58 E. faecium isolates representing mainly outbreaks and clusters of infections and colonizations among patients from 31 German hospitals. All isolates were vancomycin-resistant (vanA type). Typing results for MLVA are compared with results of macrorestriction analysis in PFGE (pulsed-field gel electrophoresis) and MLST (multi-locus sequence typing). RESULTS: All 51 but one hospital isolates from 1996-2006 were assigned to the clonal complex (CC) of epidemic-virulent, hospital-adapted lineages (MLST CC-17; MLVA CC-1) and differed from isolates of sporadic infections and colonizations (n = 7; 1991-1995) and other non-hospital origins (n = 27). Typing of all 58 hospital VRE revealed MLVA as the least discriminatory method (Simpson's diversity index 0.847) when compared to MLST (0.911) and PFGE (0.976). The two most common MLVA types MT-1 (n = 16) and MT-159 (n = 14) combined isolates of several MLST types including also major epidemic, hospital adapted, clonal types (MT-1: ST-17, ST-18, ST-280, ST-282; MT-159: ST-78, ST-192, ST-203). These data clearly indicate that non-related E. faecium could possess an identical MLVA type being especially critical when MLVA is used to elucidate supposed outbreaks with E. faecium within a single or among different hospitals. Stability of a given MLVA profile MT-12 (ST-117) during an outbreak over a period of five years was also shown. CONCLUSION: MLVA is a suitable method to assign isolates of E. faecium into distinct clonal complexes. To investigate outbreaks the current MLVA typing scheme for E. faecium does not discriminate enough and cannot be recommended as a standard superior to PFGE. PMID- 17425780 TI - Passive smoking as a risk factor of anemia in young children aged 0-35 months in Jordan. AB - BACKGROUND: Passive smoking unfavorably affects pregnancy, child birth and child health. Passive smoking associates with still-birth, premature birth as well as acute respiratory infection, asthma, disorder in red blood cell metabolism in children. This study examined the effects of passive smoking on anemia in young children in Jordan. METHODS: The analysis based on the information from 740 children aged 0-35 months that were tested for hemoglobin levels included in the 2002 Jordan Population and Family Health Survey. This study used multivariate logistic regression method to analyze the effect of passive smoking on anemia in young children in Jordan, controlling for a number of risk factors and confounding factors for anemia. RESULTS: Results indicated that independent of other risk factors and confounding factors, anemia in young children was strongly positively associated with exposure to passive smoking from both parents (OR= 2.99, p < 0.01). Severely undernourished children were at higher risk of anemia independent of passive smoking and other risk factors (OR= 5.29, p < 0.05). Children age 24-35 months, children born to mothers age 35-49, and children lived in households with a hygienic toilet facility were less likely to suffer from anemia. CONCLUSION: Passive smoking from both parents was strongly positively associated with anemia in young children in Jordan independent of other risk factors and confounding factors. The results support the importance of smoking prevention during and after pregnancy that prevent childhood anemia and others morbidities in young children. PMID- 17425781 TI - A novel bifunctional N-acetylglutamate synthase-kinase from Xanthomonas campestris that is closely related to mammalian N-acetylglutamate synthase. AB - BACKGROUND: In microorganisms and plants, the first two reactions of arginine biosynthesis are catalyzed by N-acetylglutamate synthase (NAGS) and N acetylglutamate kinase (NAGK). In mammals, NAGS produces an essential activator of carbamylphosphate synthetase I, the first enzyme of the urea cycle, and no functional NAGK homolog has been found. Unlike the other urea cycle enzymes, whose bacterial counterparts could be readily identified by their sequence conservation with arginine biosynthetic enzymes, mammalian NAGS gene was very divergent, making it the last urea cycle gene to be discovered. Limited sequence similarity between E. coli NAGS and fungal NAGK suggests that bacterial and eukaryotic NAGS, and fungal NAGK arose from the fusion of genes encoding an ancestral NAGK (argB) and an acetyltransferase. However, mammalian NAGS no longer retains any NAGK catalytic activity. RESULTS: We identified a novel bifunctional N-acetylglutamate synthase and kinase (NAGS-K) in the Xanthomonadales order of gamma-proteobacteria that appears to resemble this postulated primordial fusion protein. Phylogenetic analysis indicated that xanthomonad NAGS-K is more closely related to mammalian NAGS than to other bacterial NAGS. We cloned the NAGS-K gene from Xanthomonas campestis, and characterized the recombinant NAGS-K protein. Mammalian NAGS and its bacterial homolog have similar affinities for substrates acetyl coenzyme A and glutamate as well as for their allosteric regulator arginine. CONCLUSION: The close phylogenetic relationship and similar biochemical properties of xanthomonad NAGS-K and mammalian NAGS suggest that we have identified a close relative to the bacterial antecedent of mammalian NAGS and that the enzyme from X. campestris could become a good model for mammalian NAGS in structural, biochemical and biophysical studies. PMID- 17425782 TI - Pygo1 and Pygo2 roles in Wnt signaling in mammalian kidney development. AB - BACKGROUND: The pygopus gene of Drosophila encodes an essential component of the Armadillo (beta-catenin) transcription factor complex of canonical Wnt signaling. To better understand the functions of Pygopus-mediated canonical Wnt signaling in kidney development, targeted mutations were made in the two mammalian orthologs, Pygo1 and Pygo2. RESULTS: Each mutation deleted >80% of the coding sequence, including the critical PHD domain, and almost certainly resulted in null function. Pygo2 homozygous mutants, with rare exception, died shortly after birth, with a phenotype including lens agenesis, growth retardation, altered kidney development, and in some cases exencephaly and cleft palate. Pygo1 homozygous mutants, however, were viable and fertile, with no detectable developmental defects. Double Pygo1/Pygo2 homozygous mutants showed no apparent synergy in phenotype severity. The BAT-gal transgene reporter of canonical Wnt signaling showed reduced levels of expression in Pygo1-/-/Pygo2-/- mutants, with tissue-specific variation in degree of diminution. The Pygo1 and Pygo2 genes both showed widespread expression in the developing kidney, with raised levels in the stromal cell compartment. Confocal analysis of the double mutant kidneys showed disturbance of both the ureteric bud and metanephric mesenchyme-derived compartments. Branching morphogenesis of the ureteric bud was altered, with expanded tips and reduced tip density, probably contributing to the smaller size of the mutant kidney. In addition, there was an expansion of the zone of condensed mesenchyme capping the ureteric bud. Nephron formation, however, proceeded normally. Microarray analysis showed changed expression of several genes, including Cxcl13, Slc5a2, Klk5, Ren2 and Timeless, which represent candidate Wnt targets in kidney development. CONCLUSION: The mammalian Pygopus genes are required for normal branching morphogenesis of the ureteric bud during kidney development. Nevertheless, the relatively mild phenotype observed in the kidney, as well as other organ systems, indicates a striking evolutionary divergence of Pygopus function between mammals and Drosophila. In mammals, the Pygo1/Pygo2 genes are not absolutely required for canonical Wnt signaling in most developing systems, but rather function as quantitative transducers, or modulators, of Wnt signal intensity. PMID- 17425783 TI - Vaccination response to tetanus toxoid and 23-valent pneumococcal vaccines following administration of a single dose of abatacept: a randomized, open-label, parallel group study in healthy subjects. AB - The effect of abatacept, a selective T-cell co-stimulation modulator, on vaccination has not been previously investigated. In this open-label, single dose, randomized, parallel-group, controlled study, the effect of a single 750 mg infusion of abatacept on the antibody response to the intramuscular tetanus toxoid vaccine (primarily a memory response to a T-cell-dependent peptide antigen) and the intramuscular 23-valent pneumococcal vaccine (a less T-cell dependent response to a polysaccharide antigen) was measured in 80 normal healthy volunteers. Subjects were uniformly randomized to receive one of four treatments: Group A (control group), subjects received vaccines on day 1 only; Group B, subjects received vaccines 2 weeks before abatacept; Group C, subjects received vaccines 2 weeks after abatacept; and Group D, subjects received vaccines 8 weeks after abatacept. Anti-tetanus and anti-pneumococcal (Danish serotypes 2, 6B, 8, 9V, 14, 19F and 23F) antibody titers were measured 14 and 28 days after vaccination. While there were no statistically significant differences between the dosing groups, geometric mean titers following tetanus or pneumococcal vaccination were generally lower in subjects who were vaccinated 2 weeks after receiving abatacept, compared with control subjects. A positive response (defined as a twofold increase in antibody titer from baseline) to tetanus vaccination at 28 days was seen, however, in > or = 60% of subjects across all treatment groups versus 75% of control subjects. Similarly, over 70% of abatacept-treated subjects versus all control subjects (100%) responded to at least three pneumococcal serotypes, and approximately 25-30% of abatacept-treated subjects versus 45% of control subjects responded to at least six serotypes. PMID- 17425784 TI - Phylogenetics of Cucumis (Cucurbitaceae): cucumber (C. sativus) belongs in an Asian/Australian clade far from melon (C. melo). AB - BACKGROUND: Melon, Cucumis melo, and cucumber, C. sativus, are among the most widely cultivated crops worldwide. Cucumis, as traditionally conceived, is geographically centered in Africa, with C. sativus and C. hystrix thought to be the only Cucumis species in Asia. This taxonomy forms the basis for all ongoing Cucumis breeding and genomics efforts. We tested relationships among Cucumis and related genera based on DNA sequences from chloroplast gene, intron, and spacer regions (rbcL, matK, rpl20-rps12, trnL, and trnL-F), adding nuclear internal transcribed spacer sequences to resolve relationships within Cucumis. RESULTS: Analyses of combined chloroplast sequences (4,375 aligned nucleotides) for 123 of the 130 genera of Cucurbitaceae indicate that the genera Cucumella, Dicaelospermum, Mukia, Myrmecosicyos, and Oreosyce are embedded within Cucumis. Phylogenetic trees from nuclear sequences for these taxa are congruent, and the combined data yield a well-supported phylogeny. The nesting of the five genera in Cucumis greatly changes the natural geographic range of the genus, extending it throughout the Malesian region and into Australia. The closest relative of Cucumis is Muellerargia, with one species in Australia and Indonesia, the other in Madagascar. Cucumber and its sister species, C. hystrix, are nested among Australian, Malaysian, and Western Indian species placed in Mukia or Dicaelospermum and in one case not yet formally described. Cucumis melo is sister to this Australian/Asian clade, rather than being close to African species as previously thought. Molecular clocks indicate that the deepest divergences in Cucumis, including the split between C. melo and its Australian/Asian sister clade, go back to the mid-Eocene. CONCLUSION: Based on congruent nuclear and chloroplast phylogenies we conclude that Cucumis comprises an old Australian/Asian component that was heretofore unsuspected. Cucumis sativus evolved within this Australian/Asian clade and is phylogenetically far more distant from C. melo than implied by the current morphological classification. PMID- 17425785 TI - Stromal derived factor-1 exerts differential regulation on distinct cortical cell populations in vitro. AB - BACKGROUND: Stromal derived factor (SDF-1), an alpha chemokine, is a widely known chemoattractant in the immune system. A growing body of evidence now suggests multiple regulatory roles for SDF-1 in the developing nervous system. RESULTS: To investigate the role of SDF-1 signaling in the growth and differentiation of cortical cells, we performed numerous in vitro experiments, including gene chip and quantitative RT-PCR analysis. Using SDF-1 medium and AMD3100, a receptor antagonist, we demonstrate that the chemokine signaling regulates key events during early cortical development. First, SDF-1 signaling maintains cortical progenitors in proliferation, possibly through a mechanism involving connexin 43 mediated intercellular coupling. Second, SDF-1 signaling upregulates the differentiation of cortical GABAergic neurons, independent of sonic signaling pathway. Third, SDF-1 enables the elongation and branching of axons of cortical glutamatergic neurons. Finally, cortical cultures derived from CXCR4-/- mutants show a close parallel to AMD3100 treatment with reduced cell proliferation and differentiation of GABAergic neurons. CONCLUSION: Results from this study show that SDF-1 regulates distinct cortical cell populations in vitro. PMID- 17425786 TI - FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension. AB - BACKGROUND: Skeletal muscle atrophy can occur under many different conditions, including prolonged disuse or immobilization, cachexia, cushingoid conditions, secondary to surgery, or with advanced age. The mechanisms by which unloading of muscle is sensed and translated into signals controlling tissue reduction remains a major question in the field of musculoskeletal research. While the fibroblast growth factors (FGFs) and their receptors are synthesized by, and intimately involved in, embryonic skeletal muscle growth and repair, their role maintaining adult muscle status has not been examined. METHODS: We examined the effects of ectopic expression of FGFR1 during disuse-mediated skeletal muscle atrophy, utilizing hindlimb suspension and DNA electroporation in mice. RESULTS: We found skeletal muscle FGF4 and FGFR1 mRNA expression to be modified by hind limb suspension,. In addition, we found FGFR1 protein localized in muscle fibers within atrophying mouse muscle which appeared to be resistant to atrophy. Electroporation and ectopic expression of FGFR1 significantly inhibited the decrease in muscle fiber area within skeletal muscles of mice undergoing suspension induced muscle atrophy. Ectopic FGFR1 expression in muscle also significantly stimulated protein synthesis in muscle fibers, and increased protein degradation in weight bearing muscle fibers. CONCLUSION: These results support the theory that FGF signaling can play a role in regulation of postnatal skeletal muscle maintenance, and could offer potentially novel and efficient therapeutic options for attenuating muscle atrophy during aging, illness and spaceflight. PMID- 17425787 TI - Boys are more stunted than girls in sub-Saharan Africa: a meta-analysis of 16 demographic and health surveys. AB - BACKGROUND: Many studies in sub-Saharan Africa have occasionally reported a higher prevalence of stunting in male children compared to female children. This study examined whether there are systematic sex differences in stunting rates in children under-five years of age, and how the sex differences in stunting rates vary with household socio-economic status. METHODS: Data from the most recent 16 demographic and health surveys (DHS) in 10 sub-Saharan countries were analysed. Two separate variables for household socio-economic status (SES) were created for each country based on asset ownership and mothers' education. Quintiles of SES were constructed using principal component analysis. Sex differentials with stunting were assessed using Student's t-test, chi square test and binary logistic regressions. RESULTS: The prevalence and the mean z-scores of stunting were consistently lower amongst females than amongst males in all studies, with differences statistically significant in 11 and 12, respectively, out of the 16 studies. The pooled estimates for mean z-scores were -1.59 for boys and -1.46 for girls with the difference statistically significant (p < 0.001). The stunting prevalence was also higher in boys (40%) than in girls (36%) in pooled data analysis; crude odds ratio 1.16 (95% CI 1.12-1.20); child age and individual survey adjusted odds ratio 1.18 (95% CI 1.14-1.22). Male children in households of the poorest 40% were more likely to be stunted compared to females in the same group, but the pattern was not consistent in all studies, and evaluation of the SES/sex interaction term in relation to stunting was not significant for the surveys. CONCLUSION: In sub-Saharan Africa, male children under five years of age are more likely to become stunted than females, which might suggest that boys are more vulnerable to health inequalities than their female counterparts in the same age groups. In several of the surveys, sex differences in stunting were more pronounced in the lowest SES groups. PMID- 17425788 TI - Functional mapping of hospitals by diagnosis-dominant case-mix analysis. AB - BACKGROUND: Principles and methods for the allocation of healthcare resources among healthcare providers have long been health policy research issues in many countries. Healthcare reforms including the development of a new case-mix system, Diagnosis Procedure Combination (DPC), and the introduction of a DPC-based payment system are currently underway in Japan, and a methodology for adequately assessing the functions of healthcare providers is needed to determine healthcare resource allocations. METHODS: By two-dimensional mapping of the rarity and complexity of diagnoses for patients receiving treatment, we were able to quantitatively demonstrate differences in the functions of different healthcare service provider groups. RESULTS: On average, inpatients had diseases that were 3.6-times rarer than those seen in outpatients, while major teaching hospitals treated inpatients with diseases 3.0-times rarer on average than those seen at small hospitals. CONCLUSION: We created and evaluated a new indicator for DPC, the diagnosis-dominant case-mix system developed in Japan, whereby the system was used to assess the functions of healthcare service providers. The results suggest that it is possible to apply the case-mix system to the integrated evaluation of outpatient and inpatient healthcare services and to the appropriate allocation of healthcare resources among health service providers. PMID- 17425789 TI - Brief fruit and vegetable messages integrated within a community physical activity program successfully change behaviour. AB - BACKGROUND: Consumption of the recommended amounts of fruits and vegetables is associated with several health benefits. Currently less than 25% of the American population meets the minimum recommendation of five servings a day. In order to change this health behaviour, interventions should be based on theory and include community-wide social support. METHODS: A low intensity intervention was developed in which participants (n = 86) were randomly assigned to either the fruit and vegetable intervention (FVI) or standard control condition. The intervention was integrated into an ongoing community physical activity program and study participants were drawn from the sample of community members enrolled in the program. The FVI consisted of brief social cognitive theory-based messages delivered in nine weekly newsletters designed to improve participant outcome and self-efficacy expectations related to fruit and vegetable consumption. RESULTS: Participants in the FVI condition increased in their fruit and vegetable consumption by approximately one to one and one-third servings per day. The control condition showed no change in consumption. The effect of the intervention was enhanced when examined by the extent to which it was adopted by participants (i.e., the number of newsletters read). Those participants who read seven or more newsletters showed an increase of two servings per day. CONCLUSION: This intervention was effective at improving fruit and vegetable consumption among adults. Minimal interventions, such as newsletters, have the ability to reach large audiences and can be integrated into ongoing health promotion programs. As such, they have potential for a strong public health impact. PMID- 17425790 TI - A kingdom-specific protein domain HMM library for improved annotation of fungal genomes. AB - BACKGROUND: Pfam is a general-purpose database of protein domain alignments and profile Hidden Markov Models (HMMs), which is very popular for the annotation of sequence data produced by genome sequencing projects. Pfam provides models that are often very general in terms of the taxa that they cover and it has previously been suggested that such general models may lack some of the specificity or selectivity that would be provided by kingdom-specific models. RESULTS: Here we present a general approach to create domain libraries of HMMs for sub-taxa of a kingdom. Taking fungal species as an example, we construct a domain library of HMMs (called Fungal Pfam or FPfam) using sequences from 30 genomes, consisting of 24 species from the ascomycetes group and two basidiomycetes, Ustilago maydis, a fungal pathogen of maize, and the white rot fungus Phanerochaete chrysosporium. In addition, we include the Microsporidion Encephalitozoon cuniculi, an obligate intracellular parasite, and two non-fungal species, the oomycetes Phytophthora sojae and Phytophthora ramorum, both plant pathogens. We evaluate the performance in terms of coverage against the original 30 genomes used in training FPfam and against five more recently sequenced fungal genomes that can be considered as an independent test set. We show that kingdom-specific models such as FPfam can find instances of both novel and well characterized domains, increases overall coverage and detects more domains per sequence with typically higher bitscores than Pfam for the same domain families. An evaluation of the effect of changing E values on the coverage shows that the performance of FPfam is consistent over the range of E-values applied. CONCLUSION: Kingdom-specific models are shown to provide improved coverage. However, as the models become more specific, some sequences found by Pfam may be missed by the models in FPfam and some of the families represented in the test set are not present in FPfam. Therefore, we recommend that both general and specific libraries are used together for annotation and we find that a significant improvement in coverage is achieved by using both Pfam and FPfam. PMID- 17425791 TI - Craniopharyngioma. AB - Craniopharyngiomas are benign slow growing tumours that are located within the sellar and para sellar region of the central nervous system. The point prevalence of this tumour is approximately 2/100,000. The onset of symptoms is normally insidious with most patients at diagnosis having neurological (headaches, visual disturbances) and endocrine (growth retardation, delayed puberty) dysfunctions. Craniopharyngiomas are thought to arise from epithelial remnants of the craniopharyngeal duct or Rathke's pouch (adamantinomatous type) or from metaplasia of squamous epithelial cell rests that are remnants of the part of the stomadeum that contributed to the buccal mucosa (squamous papillary type). The neuroradiological diagnosis is mainly based on the three components of the tumour (cystic, solid and calcified) in the characteristic sellar/para sellar location. Definitive diagnosis is made following histological examination of a surgical specimen. The differential diagnosis includes other tumours in this region (pituitary adenoma), infectious or inflammatory processes (eosinophilic granuloma), vascular malformations (aneurysm) and congenital anomalies (Rathke's cleft cyst). The current treatment is gross total excision of the tumour, if there is no hypothalamic invasion or, in the presence of hypothalamic invasion, a sub-total resection with post-operative radiotherapy. Endocrine disturbances are normally permanent and need careful replacement. Overall, there is an 80% 5 year survival, though this can be associated with marked morbidity (hypothalamic dysfunction, altered neuropsychological profile). PMID- 17425792 TI - Patient risk profiles and practice variation in nonadherence to antidepressants, antihypertensives and oral hypoglycemics. AB - BACKGROUND: Many patients experience difficulties in following treatment recommendations. This study's objective is to identify nonadherence risk profiles regarding medication (antidepressants, antihypertensives, and oral hypoglycemics) from a combination of patients' socio-demographic characteristics, morbidity presented within general practice and medication characteristics. An additional objective is to explore differences in nonadherence among patients from different general practices. METHODS: Data were obtained by linkage of a Dutch general practice registration database to a dispensing registration database from the year 2001. Subjects included in the analyses were users of antidepressants (n = 4,877), antihypertensives (n = 14,219), or oral hypoglycemics (n = 2,428) and their GPs. Outcome variables were: 1) early dropout i.e., a maximum of two prescriptions and 2) refill nonadherence (in patients with 3+ prescriptions); refill adherence < 80% was considered as nonadherence. Multilevel modeling was used for analyses. RESULTS: Both early dropout and refill nonadherence were highest for antidepressants, followed by antihypertensives. Risk factors appeared medication specific and included: 1) non-western immigrants being more vulnerable for nonadherence to antihypertensives and antidepressants; 2) type of medication influencing nonadherence in both antihypertensives and antidepressants, 3) GP consultations contributing positively to adherence to antihypertensives and 4) somatic co-morbidity influencing adherence to antidepressants negatively. There was a considerable range between general practices in the proportion of patients who were nonadherent. CONCLUSION: No clear risk profiles for nonadherence could be constructed. Characteristics that are correlated with nonadherence vary across different types of medication. Moreover, both patient and prescriber influence adherence. Especially non-western immigrants need more attention with regard to nonadherence, for example by better monitoring or communication. Since it is not clear which prescriber characteristics influence adherence levels of their patients, there is need for further research into the role of the prescriber. PMID- 17425793 TI - Decision-making regarding total knee replacement surgery: a qualitative meta synthesis. AB - BACKGROUND: Knee osteoarthritis is a highly prevalent condition that can result in disability and reduced quality of life. The evidence suggests that total knee replacement surgery (TKR) is an effective intervention for patients with severe knee problems, but there is also an unmet need for this treatment in the UK. To help understand the reason for this unmet need, the aim of this study was to explore the factors that influence the decision-making process of TKR surgery by synthesising the available evidence from qualitative research on this topic. METHODS: A meta-synthesis was undertaken. This involved sevens steps: getting started, deciding what is relevant to the initial interest, reading the studies, determining how the studies are related, translating the studies into one another, synthesising translations, and finally, expressing the synthesis. Second order and third-order interpretations regarding decision-making in TKR surgery were drawn from the literature. RESULTS: Ten qualitative studies were found and are included in the synthesis. The evidence suggests that social and cultural categories of aging have shaped the expectation of knee osteoarthritis, and this in turn shapes patients' expectations of treatment options. The role of the health care professional was the strongest theme to emerge across all ten studies. Coping strategies and life context determine short and longer-term outcomes of TKR. CONCLUSION: The decision-making process regarding TKR surgery is extremely complex, as patients have weigh up numerous considerations before they can make a decision about surgery. By synthesising ten qualitative studies, we have illuminated the importance of the health care professional during this process. PMID- 17425794 TI - Protein homologous cores and loops: important clues to evolutionary relationships between structurally similar proteins. AB - BACKGROUND: To discover remote evolutionary relationships and functional similarities between proteins, biologists rely on comparative sequence analysis, and when structures are available, on structural alignments and various measures of structural similarity. The measures/scores that have most commonly been used for this purpose include: alignment length, percent sequence identity, superposition RMSD and their different combinations. More recently, we have introduced the "Homologous core structure overlap score" (HCS) and the "Loop Hausdorff Measure" (LHM). Along with these we also consider the "gapped structural alignment score" (GSAS), which was introduced earlier by other researchers. RESULTS: We analyze the performance of these and other conventional measures at the task of ranking structure neighbors by homology, and we show that the HCS, LHM, and GSAS scores display considerably improved performance over the conventional measures of sequence or structural similarity. CONCLUSION: The HCS, LHM, and GSAS scores are easily computable quantities that allow users of structure-neighbor databases to more easily identify interesting structural similarities between proteins. PMID- 17425795 TI - Facial diplegia with hyperreflexia--a mild Guillain-Barre Syndrome variant, to treat or not to treat? AB - Guillain Barre Syndrome (GBS) is readily diagnosed when the presentation is that of ascending weakness and areflexia. Atypical presentations with preserved, and at times, brisk reflexes, can be a diagnostic dilemma. We describe a patient with GBS who presented with facial diplegia and hyperreflexia on examination and discuss management options. PMID- 17425796 TI - 3D-MRI rendering of the anatomical structures related to acupuncture points of the Dai mai, Yin qiao mai and Yang qiao mai meridians within the context of the WOMED concept of lateral tension: implications for musculoskeletal disease. AB - BACKGROUND: A conceptual model of lateral muscular tension in patients presenting thyroid associated ophthalmopathy (TAO) has been recently described. Clinical improvement has been achieved by using acupuncture on points belonging to the so called extraordinary meridians. The aim of this study was to characterize the anatomical structures related to these acupuncture points by means of 3D MRI image rendering relying on external markers. METHODS: The investigation was carried out the index case patient of the lateral tension model. A licensed medical acupuncture practitioner located the following acupuncture points: 1) Yin qiao mai meridian (medial ankle): Kidney 3, Kidney 6, the plantar Kidney 6 (Nan jing description); 2) Yang qiao mai meridian (lateral ankle): Bladder 62, Bladder 59, Bladder 61, and the plantar Bladder 62 (Nan jing description); 3) Dai mai meridian (wait): Liver 13, Gall bladder 26, Gall bladder 27, Gall bladder 28, and Gall bladder 29. The points were marked by taping a nitro-glycerin capsule on the skin. Imaging was done on a Siemens Magnetom Avanto MR scanner using an array head and body coil. Mainly T1-weighted imaging sequences, as routinely used for patient exams, were used to obtain multi-slice images. The image data were rendered in 3D modus using dedicated software (Leonardo, Siemens). RESULTS: Points of the Dai mai meridian--at the level of the waist--corresponded to the obliquus externus abdominis and the obliquus internus abdominis. Points of the Yin qiao mai meridian--at the medial side of the ankle--corresponded to tendinous structures of the flexor digitorum longus as well as to muscular structures of the abductor hallucis on the foot sole. Points of the Yang qiao mai meridian--at the lateral side of the ankle--corresponded to tendinous structures of the peroneus brevis, the peroneous longus, and the lateral surface of the calcaneus and close to the foot sole to the abductor digiti minimi. CONCLUSION: This non invasive MRI investigation has revealed the anatomical relations of acupuncture points belonging to 3 of the so-called extraordinary meridians. We conclude that the clinically developed "WOMED concept of lateral tension" is related to tendino muscular structures. PMID- 17425797 TI - Modelling the evolution of the archeal tryptophan synthase. AB - BACKGROUND: Microorganisms and plants are able to produce tryptophan. Enzymes catalysing the last seven steps of tryptophan biosynthesis are encoded in the canonical trp operon. Among the trp genes are most frequently trpA and trpB, which code for the alpha and beta subunit of tryptophan synthase. In several prokaryotic genomes, two variants of trpB (named trpB1 or trpB2) occur in different combinations. The evolutionary history of these trpB genes is under debate. RESULTS: In order to study the evolution of trp genes, completely sequenced archeal and bacterial genomes containing trpB were analysed. Phylogenetic trees indicated that TrpB sequences constitute four distinct groups; their composition is in agreement with the location of respective genes. The first group consisted exclusively of trpB1 genes most of which belonged to trp operons. Groups two to four contained trpB2 genes. The largest group (trpB2_o) contained trpB2 genes all located outside of operons. Most of these genes originated from species possessing an operon-based trpB1 in addition. Groups three and four pertain to trpB2 genes of those genomes containing exclusively one or two trpB2 genes, but no trpB1. One group (trpB2_i) consisted of trpB2 genes located inside, the other (trpB2_a) of trpB2 genes located outside the trp operon. TrpA and TrpB form a heterodimer and cooperate biochemically. In order to characterise trpB variants and stages of TrpA/TrpB cooperation in silico, several approaches were combined. Phylogenetic trees were constructed for all trp genes; their structure was assessed via bootstrapping. Alternative models of trpB evolution were evaluated with parsimony arguments. The four groups of trpB variants were correlated with archeal speciation. Several stages of TrpA/TrpB cooperation were identified and trpB variants were characterised. Most plausibly, trpB2 represents the predecessor of the modern trpB gene, and trpB1 evolved in an ancestral bacterium. CONCLUSION: In archeal genomes, several stages of trpB evolution, TrpA/TrpB cooperation, and operon formation can be observed. Thus, archeal trp genes may serve as a model system for studying the evolution of protein-protein interactions and operon formation. PMID- 17425798 TI - European citizens' use of E-health services: a study of seven countries. AB - BACKGROUND: European citizens are increasingly being offered Internet health services. This study investigated patterns of health-related Internet use, its consequences, and citizens' expectations about their doctors' provision of e health services. METHODS: Representative samples were obtained from the general populations in Norway, Denmark, Germany, Greece, Poland, Portugal and Latvia. The total sample consisted of 7934 respondents. Interviews were conducted by telephone. RESULTS: 44 % of the total sample, 71 % of the Internet users, had used the Internet for health purposes. Factors that positively affected the use of Internet for health purposes were youth, higher education, white-collar or no paid job, visits to the GP during the past year, long-term illness or disabilities, and a subjective assessment of one's own health as good. Women were the most active health users among those who were online. One in four of the respondents used the Internet to prepare for or follow up doctors' appointments. Feeling reassured after using the Internet for health purposes was twice as common as experiencing anxieties. When choosing a new doctor, more than a third of the sample rated the provision of e-health services as important. CONCLUSION: The users of Internet health services differ from the general population when it comes to health and demographic variables. The most common way to use the Internet in health matters is to read information, second comes using the net to decide whether to see a doctor and to prepare for and follow up on doctors' appointments. Hence, health-related use of the Internet does affect patients' use of other health services, but it would appear to supplement rather than to replace other health services. PMID- 17425799 TI - Molecular and cellular characterization of ABCG2 in the prostate. AB - BACKGROUND: Identification and characterization of the prostate stem cell is important for understanding normal prostate development and carcinogenesis. The flow cytometry-based side population (SP) technique has been developed to isolate putative adult stem cells in several human tissue types including the prostate. This phenotype is mainly mediated by the ATP-binding cassette membrane transporter ABCG2. METHODS: Immunolocalization of ABCG2 was performed on normal prostate tissue obtained from radical prostatectomies. Normal human prostate SP cells and ABCG2+ cells were isolated and gene expression was determined with DNA array analysis and RT-PCR. Endothelial cells were removed by pre-sorting with CD31. RESULTS: ABCG2 positive cells were localized to the prostate basal epithelium and endothelium. ABCG2+ cells in the basal epithelium constituted less than 1% of the total basal cell population. SP cells constituted 0.5-3% of the total epithelial fraction. The SP transcriptome was essentially the same as ABCG2+ and both populations expressed genes indicative of a stem cell phenotype, however, the cells also expressed many genes in common with endothelial cells. CONCLUSION: These results provide gene expression profiles for the prostate SP and ABCG2+ cells that will be critical for studying normal development and carcinogenesis, in particular as related to the cancer stem cell concept. PMID- 17425800 TI - Does vancomycin prescribing intervention affect vancomycin-resistant enterococcus infection and colonization in hospitals? A systematic review. AB - BACKGROUND: Vancomycin resistant enterococcus (VRE) is a major cause of nosocomial infections in the United States and may be associated with greater morbidity, mortality, and healthcare costs than vancomycin-susceptible enterococcus. Current guidelines for the control of VRE include prudent use of vancomycin. While vancomycin exposure appears to be a risk factor for VRE acquisition in individual patients, the effect of vancomycin usage at the population level is not known. We conducted a systematic review to determine the impact of reducing vancomycin use through prescribing interventions on the prevalence and incidence of VRE colonization and infection in hospitals within the United States. METHODS: To identify relevant studies, we searched three electronic databases, and hand searched selected journals. Thirteen studies from 12 articles met our inclusion criteria. Data were extracted and summarized for study setting, design, patient characteristics, types of intervention(s), and outcome measures. The relative risk, 95% confidence interval, and p-value associated with change in VRE acquisition pre- and post-vancomycin prescription interventions were calculated and compared. Heterogeneity in study results was formally explored by stratified analysis. RESULTS: No randomized clinical trials on this topic were found. Each of the 13 included studies used a quasi experimental design of low hierarchy. Seven of the 13 studies reported statistically significant reductions in VRE acquisition following interventions, three studies reported no significant change, and three studies reported increases in VRE acquisition, one of which reported statistical significance. Results ranged from a reduction of 82.5% to an increase of 475%. Studies of specific wards, which included sicker patients, were more likely to report positive results than studies of an entire hospital including general inpatients (Fisher's exact test 0.029). The type of intervention, endemicity status, type of study design, and the duration of intervention were not found to significantly modify the results. Among the six studies that implemented vancomycin reduction strategies as the sole intervention, two of six (33%) found a significant reduction in VRE colonization and/or infection. In contrast, among studies implementing additional VRE control measures, five of seven (71%) reported a significant reduction. CONCLUSION: It was not possible to conclusively determine a potential role for vancomycin usage reductions in controlling VRE colonization and infection in hospitals in the United States. The effectiveness of such interventions and their sustainability remains poorly defined because of the heterogeneity and quality of studies. Future research using high-quality study designs and implementing vancomycin as the sole intervention are needed to answer this question. PMID- 17425802 TI - Core histone genes of Giardia intestinalis: genomic organization, promoter structure, and expression. AB - BACKGROUND: Giardia intestinalis is a protist found in freshwaters worldwide, and is the most common cause of parasitic diarrhea in humans. The phylogenetic position of this parasite is still much debated. Histones are small, highly conserved proteins that associate tightly with DNA to form chromatin within the nucleus. There are two classes of core histone genes in higher eukaryotes: DNA replication-independent histones and DNA replication-dependent ones. RESULTS: We identified two copies each of the core histone H2a, H2b and H3 genes, and three copies of the H4 gene, at separate locations on chromosomes 3, 4 and 5 within the genome of Giardia intestinalis, but no gene encoding a H1 linker histone could be recognized. The copies of each gene share extensive DNA sequence identities throughout their coding and 5' noncoding regions, which suggests these copies have arisen from relatively recent gene duplications or gene conversions. The transcription start sites are at triplet A sequences 1-27 nucleotides upstream of the translation start codon for each gene. We determined that a 50 bp region upstream from the start of the histone H4 coding region is the minimal promoter, and a highly conserved 15 bp sequence called the histone motif (him) is essential for its activity. The Giardia core histone genes are constitutively expressed at approximately equivalent levels and their mRNAs are polyadenylated. Competition gel-shift experiments suggest that a factor within the protein complex that binds him may also be a part of the protein complexes that bind other promoter elements described previously in Giardia. CONCLUSION: In contrast to other eukaryotes, the Giardia genome has only a single class of core histone genes that encode replication-independent histones. Our inability to locate a gene encoding the linker histone H1 leads us to speculate that the H1 protein may not be required for the compaction of Giardia's small and gene-rich genome. PMID- 17425803 TI - BranchClust: a phylogenetic algorithm for selecting gene families. AB - BACKGROUND: Automated methods for assembling families of orthologous genes include those based on sequence similarity scores and those based on phylogenetic approaches. The first are easy to automate but usually they do not distinguish between paralogs and orthologs or have restriction on the number of taxa. Phylogenetic methods often are based on reconciliation of a gene tree with a known rooted species tree; a limitation of this approach, especially in case of prokaryotes, is that the species tree is often unknown, and that from the analyses of single gene families the branching order between related organisms frequently is unresolved. RESULTS: Here we describe an algorithm for the automated selection of orthologous genes that recognizes orthologous genes from different species in a phylogenetic tree for any number of taxa. The algorithm is capable of distinguishing complete (containing all taxa) and incomplete (not containing all taxa) families and recognizes in- and outparalogs. The BranchClust algorithm is implemented in Perl with the use of the BioPerl module for parsing trees and is freely available at http://bioinformatics.org/branchclust. CONCLUSION: BranchClust outperforms the Reciprocal Best Blast hit method in selecting more sets of putatively orthologous genes. In the test cases examined, the correctness of the selected families and of the identified in- and outparalogs was confirmed by inspection of the pertinent phylogenetic trees. PMID- 17425804 TI - Receptor for advanced glycation end products Glycine 82 Serine polymorphism and risk of cardiovascular events in rheumatoid arthritis. AB - Patients with rheumatoid arthritis (RA) are at risk of excess mortality, predominantly owing to cardiovascular (CV) events. The receptor for advanced glycation end products (RAGE) has been implicated in the perpetuation of the chronic inflammatory response in vascular disease. A Gly82-->Ser polymorphism in the RAGE gene, which is associated with enhanced RAGE signaling, is present more frequently in patients with RA than the general population. To investigate whether RAGE Gly82-->Ser polymorphism is associated with CV events in RA, we examined CV events, CV risk factors, features of RA and RAGE Gly82-->Ser polymorphism in 232 patients with RA attending a tertiary referral hospital. CV events, the duration and severity of RA, and risk factors for CV disease were determined using patient questionnaires, chart review, laboratory analysis and radiographs. DNA was typed for HLA-DRB1 genes and RAGE Gly82-->Ser polymorphism. The RAGE Ser82 allele, which is in linkage disequilibrium with the RA susceptibility allele HLA-DRB1*0401, was carried by 20% of patients. More than 20% of the cohort had suffered a vascular event; a shorter duration of RA, but not the RAGE genotype, was significantly associated with CV events. However, a history of statin use was protective. Thus, the RAGE Ser82 allele, associated with enhanced RAGE signaling, does not predispose to CV events in RA. However, treatment of hyperlipidemia with statins reduces the probability of a CV event. PMID- 17425801 TI - The transcriptome analysis of early morphogenesis in Paracoccidioides brasiliensis mycelium reveals novel and induced genes potentially associated to the dimorphic process. AB - BACKGROUND: Paracoccidioides brasiliensis is a human pathogen with a broad distribution in Latin America. The fungus is thermally dimorphic with two distinct forms corresponding to completely different lifestyles. Upon elevation of the temperature to that of the mammalian body, the fungus adopts a yeast-like form that is exclusively associated with its pathogenic lifestyle. We describe expressed sequence tags (ESTs) analysis to assess the expression profile of the mycelium to yeast transition. To identify P. brasiliensis differentially expressed sequences during conversion we performed a large-scale comparative analysis between P. brasiliensis ESTs identified in the transition transcriptome and databases. RESULTS: Our analysis was based on 1107 ESTs from a transition cDNA library of P. brasiliensis. A total of 639 consensus sequences were assembled. Genes of primary metabolism, energy, protein synthesis and fate, cellular transport, biogenesis of cellular components were represented in the transition cDNA library. A considerable number of genes (7.51%) had not been previously reported for P. brasiliensis in public databases. Gene expression analysis using in silico EST subtraction revealed that numerous genes were more expressed during the transition phase when compared to the mycelial ESTs 1. Classes of differentially expressed sequences were selected for further analysis including: genes related to the synthesis/remodeling of the cell wall/membrane. Thirty four genes from this family were induced. Ten genes related to signal transduction were increased. Twelve genes encoding putative virulence factors manifested increased expression. The in silico approach was validated by northern blot and semi-quantitative RT-PCR. CONCLUSION: The developmental program of P. brasiliensis is characterized by significant differential positive modulation of the cell wall/membrane related transcripts, and signal transduction proteins, suggesting the related processes important contributors to dimorphism. Also, putative virulence factors are more expressed in the transition process suggesting adaptation to the host of the yeast incoming parasitic phase. Those genes provide ideal candidates for further studies directed at understanding fungal morphogenesis and its regulation. PMID- 17425805 TI - RNA mutagenesis yields highly diverse mRNA libraries for in vitro protein evolution. AB - BACKGROUND: In protein drug development, in vitro molecular optimization or protein maturation can be used to modify protein properties. One basic approach to protein maturation is the introduction of random DNA mutations into the target gene sequence to produce a library of variants that can be screened for the preferred protein properties. Unfortunately, the capability of this approach has been restricted by deficiencies in the methods currently available for random DNA mutagenesis and library generation. Current DNA based methodologies generally suffer from nucleotide substitution bias that preferentially mutate particular base pairs or show significant bias with respect to transitions or transversions. In this report, we describe a novel RNA-based random mutagenesis strategy that utilizes Qbeta replicase to manufacture complex mRNA libraries with a mutational spectrum that is close to the ideal. RESULTS: We show that Qbeta replicase generates all possible base substitutions with an equivalent preference for mutating A/T or G/C bases and with no significant bias for transitions over transversions. To demonstrate the high diversity that can be sampled from a Qbeta replicase-generated mRNA library, the approach was used to evolve the binding affinity of a single domain VNAR shark antibody fragment (12Y-2) against malarial apical membrane antigen-1 (AMA-1) via ribosome display. The binding constant (KD) of 12Y-2 was increased by 22-fold following two consecutive but discrete rounds of mutagenesis and selection. The mutagenesis method was also used to alter the substrate specificity of beta-lactamase which does not significantly hydrolyse the antibiotic cefotaxime. Two cycles of RNA mutagenesis and selection on increasing concentrations of cefotaxime resulted in mutants with a minimum 10,000 fold increase in resistance, an outcome achieved faster and with fewer overall mutations than in comparable studies using other mutagenesis strategies. CONCLUSION: The RNA based approach outlined here is rapid and simple to perform and generates large, highly diverse populations of proteins, each differing by only one or two amino acids from the parent protein. The practical implications of our results are that suitable improved protein candidates can be recovered from in vitro protein evolution approaches using significantly fewer rounds of mutagenesis and selection, and with little or no collateral damage to the protein or its mRNA. PMID- 17425806 TI - p42MAPK-mediated phosphorylation of xEIAP/XLX in Xenopus cytostatic factor arrested egg extracts. AB - BACKGROUND: BIR family proteins are evolutionarily conserved anti-apoptotic molecules. One member of Xenopus BIR family proteins, xEIAP/XLX, is a weak apoptosis inhibitor and rapidly degraded in a cell-free apoptotic execution system derived from interphase egg extracts. However, unfertilized eggs are naturally arrested at the metaphase of meiosis II by the concerted activities of Mos-MEK-p42MAPK-p90Rsk kinase cascade (cytostatic factor pathway) and many mitotic kinases. Previous studies suggest that cytostatic factor-arrested egg extracts are more resistant to spontaneous apoptosis than interphase egg extracts in a p42MAPK-dependent manner. We tested whether xEIAP/XLX might be phosphorylated in cytostatic factor-arrested egg extracts, and also examined whether xEIAP/XLX could be functionally regulated by phosphorylation. RESULTS: We found that p42MAPK was the major kinase phosphorylating xEIAP/XLX in cytostatic factor-arrested egg extracts, and three Ser residues (Ser 235/251/254) were identified as p42MAPK-mediated phosphorylation sites. We characterized the behaviors of various xEIAP/XLX mutants that could not be phosphorylated by p42MAPK. However, neither protein stability nor anti-apoptotic ability of xEIAP/XLX was significantly altered by the substitution of Ser with either Ala or Asp at these three sites. CONCLUSION: xEIAP/XLX is physiologically phosphorylated by p42MAPK in Xenopus unfertilized eggs. However, this protein may not serve as an essential mediator of p42MAPK-dependent anti-apoptotic activity. PMID- 17425807 TI - Expression profiling of genes regulated by TGF-beta: differential regulation in normal and tumour cells. AB - BACKGROUND: TGF-beta is one of the key cytokines implicated in various disease processes including cancer. TGF-beta inhibits growth and promotes apoptosis in normal epithelial cells and in contrast, acts as a pro-tumour cytokine by promoting tumour angiogenesis, immune-escape and metastasis. It is not clear if various actions of TGF-beta on normal and tumour cells are due to differential gene regulations. Hence we studied the regulation of gene expression by TGF-beta in normal and cancer cells. RESULTS: Using human 19 K cDNA microarrays, we show that 1757 genes are exclusively regulated by TGF-beta in A549 cells in contrast to 733 genes exclusively regulated in HPL1D cells. In addition, 267 genes are commonly regulated in both the cell-lines. Semi-quantitative and real-time qRT PCR analysis of some genes agrees with the microarray data. In order to identify the signalling pathways that influence TGF-beta mediated gene regulation, we used specific inhibitors of p38 MAP kinase, ERK kinase, JNK kinase and integrin signalling pathways. The data suggest that regulation of majority of the selected genes is dependent on at least one of these pathways and this dependence is cell type specific. Interestingly, an integrin pathway inhibitor, RGD peptide, significantly affected TGF-beta regulation of Thrombospondin 1 in A549 cells. CONCLUSION: These data suggest major differences with respect to TGF-beta mediated gene regulation in normal and transformed cells and significant role of non-canonical TGF-beta pathways in the regulation of many genes by TGF-beta. PMID- 17425808 TI - A network perspective on the topological importance of enzymes and their phylogenetic conservation. AB - BACKGROUND: A metabolic network is the sum of all chemical transformations or reactions in the cell, with the metabolites being interconnected by enzyme catalyzed reactions. Many enzymes exist in numerous species while others occur only in a few. We ask if there are relationships between the phylogenetic profile of an enzyme, or the number of different bacterial species that contain it, and its topological importance in the metabolic network. Our null hypothesis is that phylogenetic profile is independent of topological importance. To test our null hypothesis we constructed an enzyme network from the KEGG (Kyoto Encyclopedia of Genes and Genomes) database. We calculated three network indices of topological importance: the degree or the number of connections of a network node; closeness centrality, which measures how close a node is to others; and betweenness centrality measuring how frequently a node appears on all shortest paths between two other nodes. RESULTS: Enzyme phylogenetic profile correlates best with betweenness centrality and also quite closely with degree, but poorly with closeness centrality. Both betweenness and closeness centralities are non-local measures of topological importance and it is intriguing that they have contrasting power of predicting phylogenetic profile in bacterial species. We speculate that redundancy in an enzyme network may be reflected by betweenness centrality but not by closeness centrality. We also discuss factors influencing the correlation between phylogenetic profile and topological importance. CONCLUSION: Our analysis falsifies the hypothesis that phylogenetic profile of enzymes is independent of enzyme network importance. Our results show that phylogenetic profile correlates better with degree and betweenness centrality, but less so with closeness centrality. Enzymes that occur in many bacterial species tend to be those that have high network importance. We speculate that this phenomenon originates in mechanisms driving network evolution. Closeness centrality reflects phylogenetic profile poorly. This is because metabolic networks often consist of distinct functional modules and some are not in the centre of the network. Enzymes in these peripheral parts of a network might be important for cell survival and should therefore occur in many bacterial species. They are, however, distant from other enzymes in the same network. PMID- 17425809 TI - E-cadherin expression and bromodeoxyuridine incorporation during development of ovarian inclusion cysts in age-matched breeder and incessantly ovulated CD-1 mice. AB - BACKGROUND: Female CD-1/Swiss Webster mice subjected to incessant ovulation for 8 months and 12-month breeder mice both developed ovarian inclusion cysts similar to serous cystadenomas. The majority of cysts appeared to be dilated rete ovarii tubules, but high ovulation number resulted in more cortical inclusion cysts. We hypothesized that comparison of inclusion cyst pathology in animals of the same age, but with differences in total lifetime ovulation number, might allow us to determine distinguishing characteristics of the two types of cyst. METHODS: Ovaries from breeder mice (BR) or females subjected to incessant ovulation (IO) were compared at 6-, 9- and 12-months of age. Ovaries were serially sectioned and cysts characterized with regard to location and histology, E-cadherin immunoreactivity and rates of BrdU incorporation. RESULTS: Inclusion cysts developed with age in BR and IO ovaries. The majority of cysts were connected to the ovarian hilus. Two cortical inclusion cysts were observed in ten IO ovaries and one in ten BR ovaries. Low or no E-cadherin immuno-staining was seen in the OSE of all mice studied. Conversely, strong membrane immuno-staining was observed in rete ovarii epithelial cells. Variable E-cadherin immunoreactivity was seen in cells of hilar inclusion cysts, with strong staining observed in cuboidal ciliated cells and little or no staining in flat epithelial cells. Two of the three cortical cysts contained papillae, which showed E-cadherin immuno-staining at the edge of cells. However hilar and cortical cysts were not distinguishable by morphology, cell type or E-cadherin immunoreactivity. BrdU incorporation in cyst cells (1.4% [95% CI: 1.0 to 2.1]) was greater than in OSE (0.7% [95% CI: 0.4 to 1.2]) and very few BrdU-labeled cells were observed in rete ovarii at any age. Incessant ovulation significantly increased BrdU incorporation in OSE of older animals. CONCLUSION: These experiments confirm ovarian inclusion cysts develop with age in the CD-1 mouse strain, irrespective of total ovulation burden. We conclude longer periods of incessant ovulation do not lead to significant changes in inclusion cyst formation or steroidogenesis in CD-1 mice and inclusion cyst type can not be distinguished by morphology, cell proliferation rate or E cadherin immunoreactivity. PMID- 17425810 TI - Physician burnout: a time for healing. PMID- 17425811 TI - Rupture of the primigravid uterus: a review of the literature. AB - Uterine rupture is a catastrophic obstetric complication, associated with high rates of perinatal morbidity and mortality. The most common risk factor is previous uterine surgery, and most cases of uterine rupture occur in women with a previous cesarean delivery. Traditionally, the primigravid uterus has been considered almost immune to spontaneous rupture. In fact, although spontaneous rupture of the primigravid uterus is indeed a very rare event, a number of such cases have been reported recently. Prompt recognition of uterine rupture and expeditious recourse to laparotomy are critical in influencing perinatal and maternal morbidity. Not all uterine ruptures present with the typical clinical picture of abdominal pain, hypovolemia, vaginal bleeding, and fetal compromise. Therefore, it is important to maintain a high index of suspicion for uterine rupture in women presenting with some, or all, of these features, regardless of parity. Here we provide a systematic review of cases of spontaneous uterine rupture in primigravid women reported in the literature to date. Clinical presentation, differential diagnosis, common etiological factors, complication rates, and appropriate management of this rare obstetric event are discussed. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to recall that uterine rupture in a primigravida is a rare event, without typical signs and symptoms, and explain that the morbidity and mortality of the mother and child is directly related to a high index of suspicion and prompt treatment by the clinician. PMID- 17425812 TI - The role of oxidative stress in spontaneous abortion and recurrent pregnancy loss: a systematic review. AB - Human reproduction is not considered a highly efficient biological process. Before the end of the first trimester, 30%-50% of conceptions end in spontaneous abortion. Most losses occur at the time of implantation. 15%-20% of clinical pregnancies end in spontaneous abortions. Recurrent pregnancy loss is a frustrating clinical problem both for clinicians and patients. Recurrent pregnancy loss affects 0.5%-3% of women in the reproductive age group, and between 50%-60% of recurrent pregnancy losses are idiopathic. Oxidative stress induced damage has been hypothesized to play a role in spontaneous abortion, idiopathic recurrent pregnancy loss, hydatidiform mole, defective embryogenesis, and drug-induced teratogenicity. Some studies implicate systemic and placental oxidative stress in the pathophysiology of abortion and recurrent pregnancy loss. Oxidant-induced endothelial damage, impaired placental vascularization and immune malfunction have all been proposed to play a role in the pathophysiology of idiopathic recurrent pregnancy loss. Oxidative stress-induced placental dysfunction may be a common cause of the multifactorial and polygenic etiologies of abortion, recurrent pregnancy loss, defective embryogenesis, hydatidiform mole, and drug-induced teratogenic effects. Oxidative stress-induced modification of phospholipids has been linked to the formation of antiphospholipid antibodies in the antiphospholipid syndrome. The objective of this review was to examine the association between oxidative stress, spontaneous abortion and recurrent pregnancy loss, based on the published literature. We conducted an extensive literature search utilizing the databases of Medline, CINAHL, and Cochrane from 1986 to 2005. The following keywords were used: oxidative stress, abortion, recurrent pregnancy loss, reactive oxygen species, antioxidants, fetal development, and embryopathies. We conducted an electronic search, as well as a manual search of cross-references. We have included all studies in the English language found in the literature focusing on oxidative stress and its association with abortions, recurrent pregnancy loss and drug-induced teratogenicity. The role of antioxidant vitamins for primary prevention of oxidative stress-induced pathologies needs to be investigated further. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to state that the causes of spontaneous and recurrent abortion are multifaceted, however, some of the causes may be preventable and also explain that the role of oxidative stress during pregnancy and adverse pregnancy outcomes has a basis in pathophysiology, although the role of oxidative stress and the treatment of oxidative stress during or before pregnancy remains speculative. PMID- 17425813 TI - Urethral pain syndrome and its management. AB - Urethral pain syndrome is occurrence of persistent or recurrent episodic urethral pain usually on voiding with daytime frequency and nocturia, in the absence of proven infection or other obvious pathology. This is a condition of uncertain etiology. The objective of this article is to review etiology, diagnosis, and management of urethral pain syndrome. Since there is paucity of information on randomized trials, search of published literature has been made using keywords such as "urethral pain syndrome", "painful bladder syndrome", "urethral stenosis", and "lower urinary tract symptoms". Urethral pain syndrome is a disease of ambiguous etiology. Diagnosis is mainly based upon symptoms, and investigations are aimed to exclude other conditions affecting lower urinary tract. Various modalities of treatment including antibiotics, alpha-blockers, acupuncture, and laser therapy have been proved successful. Psychological support is very important in this group of women. Management requires multidisciplinary approach, and treatment at its best is by trial and error. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to explain that the urethral pain syndrome (UPS) has specific signs and symptoms without specific responses to a variety of treatment options and recall that treatment may require a multidisciplinary approach and a lot of sensitivity by the physician. PMID- 17425814 TI - Monitoring with EEG entropy decreases propofol requirement and maintains cardiovascular stability during induction of anaesthesia in elderly patients. AB - BACKGROUND AND OBJECTIVES: Electroencephalographic entropy is used to measure the degree of brain hypnosis and anaesthesia depth. Two parameters are displayed in the monitor, state entropy (SE) and response entropy (RE). Ageing leads to reduction in liver mass as well as hepatic blood flow, which decreases clearance of propofol and increases the risk of cardiovascular adverse effects. The aim of this study is to demonstrate the effect of electroencephalographic entropy on propofol requirement and haemodynamic parameters during induction of anaesthesia in elderly patients. METHODS: We studied 72 elderly patients. Standard monitoring was performed for all patients together with entropy monitor. Patients were allocated randomly either to the control group, which were given the recommended induction dose of propofol, or to the entropy group which was induced with propofol based on entropy reading where the end-point was SE 50 and SE-RE difference less than 10. Propofol induction doses and haemodynamic changes were recorded. Anaesthesia was maintained using sevoflurane and O2 air mixture. RESULTS: After induction of anaesthesia, the systolic, diastolic, mean arterial pressure, RE and SE were significantly lower in the control group (P value < 0.05). Total dose of propofol and the dose kg-1 were significantly reduced by 37.1% and 31.8%, respectively, in the entropy group (P value < 0.01). CONCLUSION: The use of electroencephalographic entropy during induction of anaesthesia in elderly patients reduces propofol requirements and maintains cardiovascular stability. PMID- 17425815 TI - Survey on intraoperative temperature management in Europe. AB - BACKGROUND AND OBJECTIVES: Inadvertent perioperative hypothermia causes serious morbidity in surgical patients. However, recent reports suggest that patients might still be hypothermic after elective surgery. We thus surveyed intraoperative temperature monitoring and management practices in Europe. METHODS: Postal survey of 801 representative hospitals from 17 European countries on the same day. The questions addressed the number of surgical procedures and type of anaesthesia performed, mode and site of temperature monitoring and method of patient warming. Mean and standard error of the mean or count and percentage were calculated. The t-test or contingency table analysis with the Fisher's exact test were used. RESULTS: Eight thousand and eighty-three surgical procedures were assessed from 316 responding hospitals (39.4%). Overall, patient temperature monitored in 19.4% and 38.5% of the patients were actively warmed. Under general anaesthesia, body temperature was monitored in 25% and during regional anaesthesia in 6%, P = 0.0005. Nasopharyngeal temperature was most often taken under general anaesthesia, while tympanic temperature was preferred during regional anaesthesia. Under general anaesthesia, 43% of patients were actively warmed as compared to 28% with regional anaesthesia, P = 0.0005. Forced-air warming was the method of choice for both general and regional anaesthesia. CONCLUSIONS: Intraoperative temperature monitoring is still uncommon and hence active patient warming is not a standard of care in Europe. Awareness of perioperative hypothermia is critical to its prevention, and thus temperature monitoring is a pre-requisite. The objective is to maintain normothermia in patients throughout surgery. A European practice guideline for perioperative patient temperature management is warranted. PMID- 17425816 TI - Assessment of the clinical relevance of quantitative sensory testing with Von Frey monofilaments in patients with allodynia and neuropathic pain. A pilot study. AB - BACKGROUND: Allodynia is a common and disabling symptom in many patients with neuropathic pain. Whereas quantification of pain mostly depends on subjective pain reports, allodynia can also be measured objectively with quantitative sensory testing. In this pilot study, we investigated the clinical relevance of quantitative sensory testing with Von Frey monofilaments in patients with allodynia as a consequence of a neuropathic pain syndrome, by means of correlating subjective pain scores with pain thresholds obtained with quantitative sensory testing. METHODS: During a 4-week trial, we administered a cannabis extract to 17 patients with allodynia. We quantified the severity of the allodynia with Von Frey monofilaments before, during and after the patients finished the trial. We also asked the patients to rate their pain on a numeric rating scale at these three moments. RESULTS: We found that most of the effect of the cannabis occurred in the last 2 weeks of the trial. In this phase, we observed that the pain thresholds, as measured with Von Frey monofilaments, were inversely correlated with a decrease of the perceived pain intensity. CONCLUSION: These preliminary findings indicate clinical relevance of quantitative sensory testing with Von Frey monofilaments in the quantification of allodynia in patients with neuropathic pain, although confirmation of our data is still required in further studies to position this method of quantitative sensory testing as a valuable tool, for example, in the evaluation of therapeutic interventions for neuropathic pain. PMID- 17425817 TI - Antiemetic effect of ondansetron 0.2 mg mL-1 in PCA morphine solution. AB - BACKGROUND AND OBJECTIVES: To study the effect of 0.2 mg mL-1 of ondansetron added to morphine patient-controlled analgesia solution after a 4 mg loading dose on the incidence and severity of postoperative nausea and vomiting. METHODS: One hundred and sixty patients scheduled for elective surgery, between 18 and 65 yr old, were studied. Patients who smoked, received antiemetics and hormonal therapy, had a history of motion sickness or gastrointestinal disease, a body mass index >35 or menstruation at the time of the study were excluded. Patients were assigned to the ondansetron and control groups by block randomization. At the end of anaesthesia, all patients received 4 mg of ondansetron intravenously and the same patient-controlled analgesia regimen of morphine. The ondansetron group (n = 80) received 0.2 mg of ondansetron per 1 mg of morphine. The nausea score, vomiting score and the requested ondansetron dose were evaluated at 1, 2, 6, 12 and 24 h. Patient-satisfaction for nausea/vomiting was recorded at the end of the study. RESULTS: Patient characteristics and cumulative morphine consumption were similar but ondansetron group had higher pain scores (P = 0.006). The ondansetron group had a lower nausea and vomiting scores, and more patients were free from nausea and vomiting than the control group (41 vs. 26, respectively, P = 0.025). The ondansetron group had fewer cumulative ondansetron doses than the control group and better patient satisfaction than the control group (P < 0.05). CONCLUSION(S): Ondansetron 4 mg plus 0.2 mg mL-1 given with PCA morphine can reduce nausea and vomiting thus improving patient satisfaction. PMID- 17425818 TI - Formation of thrombus in a native aortic sinus of Valsalva after palliation of hypoplastic left heart syndrome. AB - An eight-month-old girl with hypoplastic left heart syndrome, who underwent a modified Norwood operation at the age of two days, and a Hemifontan operation at five months of age, had severely impaired ventricular function and new electrocardiographic changes. Coronary angiography demonstrated a small adherent thrombus in the non-coronary sinus of Valsalva of the native aortic root, which may be the cause of unexplained ventricular dysfunction. PMID- 17425819 TI - Past semen donors' views about the use of a voluntary contact register. AB - This paper reports on qualitative data from a UK Department of Health-funded telephone interview study of 32 men who donated sperm samples at King's College Hospital, London between 1988 and 2002. The study considers the donors' attitudes and views about a proposed voluntary information exchange and contact register. The results suggest that semen donors continue to manage thoughts and feelings that arise from donation throughout their lifetimes, linked to their philosophical beliefs and/or evolving personal and social experiences. Some donors supported the register proposal even if they anticipated that there might be a personal cost to themselves. Several saw contact with donor offspring and/or knowledge about the outcome of their donation as potentially satisfying, indicating that it is possible to retain an interest in those to whom one is genetically related regardless of social contact. There was strong support among donors to have: (i) some control over information release and contact and (ii) access to ongoing information, advice or support from professionals who are skilled and experienced in search and reunion services. Some donors would be willing to be approached for information or contact even if they did not register. The findings indicate important policy and practice messages about the ongoing needs of donors. PMID- 17425820 TI - Antioxidants to reduce sperm DNA fragmentation: an unexpected adverse effect. AB - Reactive oxygen species (ROS) have a negative impact on sperm DNA, leading to the formation of oxidative products such as 8-oxo-7,8-dihydroxyguanosine. This compound causes fragmentation and, thus, has a mutagenic effect. Patient treatment with oral antioxidant vitamins is, therefore, standard practice for male infertility, in an attempt to decrease formation of ROS and improve fertility. In this study, the DNA fragmentation index and the degree of sperm decondensation were measured using the sperm chromatin structure assay before and after 90 days treatment with antioxidant vitamins associated with zinc and selenium. Antioxidant treatment led to a decrease in sperm DNA fragmentation ( 19.1%, P < 0.0004), suggesting that at least part of the decay was linked to ROS. However, it also led to an unexpected negative effect: an increase in sperm decondensation with the same order of magnitude (+22.8%, P < 0.0009). The opening of interchain disulphide bridges in protamines may explain this aspect, as antioxidant vitamins, especially vitamin C, are able to open the cystin net, thus interfering with paternal gene activity during preimplantation development. This observation might explain the discrepancy observed concerning the role of these antioxidant treatments in improving male fertility. PMID- 17425821 TI - Effects of failed oocyte activation and sperm protamine deficiency on fertilization post-ICSI. AB - Sperm premature chromosomal condensation (PCC) has been associated with failed fertilization. Previous studies suggest that protamine deficiency or failed oocyte activation may make spermatozoa prone to PCC. However, it is not clear which of these two factors has a more profound effect on fertilization failure. In order to distinguish between these two phenomena, oocytes that failed to fertilize after intracytoplasmic sperm injection (ICSI) were artificially activated and the association between protamine deficiency and PCC was evaluated in the remaining oocytes that failed to fertilize. The results of this study reveal that after artificial activation, fertilization rate post-ICSI increased from 59.95 to 87.7% and PCC spermatozoa appeared to be present in over 50% of the remaining oocytes that failed to fertilize. The percentage of sperm PCC was significantly higher in protamine deficient samples, thus suggesting that after failed oocyte activation, sperm PCC induced by protamine deficiency may be considered as an alternative cause of failed fertilization post-ICSI. Furthermore, the results of this study did not show any correlation between pronuclei size asynchrony and protamine deficiency. PMID- 17425822 TI - How should the clinical efficiency of oocyte cryopreservation be measured? AB - Clinical application of oocyte cryopreservation may be in the context of fertility preservation for women about to undergo cytotoxic therapies or may be as an alternative to embryo cryopreservation in routine assisted reproduction. The clinical efficiency of oocyte cryopreservation will be a consequence of the cumulative impact of pre-freeze oocyte quality, postthaw survival and subcellular effects of cryopreservation protocols, which impact on early embryo quality and post-transfer viability, together with the degree of selection which is applied to the available biological material. Any valid assessment must include reference to all the above aspects, particularly when comparing cryopreserved oocytes with non-frozen controls or cryopreserved embryos. Cumulative pregnancy rates from oocyte collections may provide the most relevant index of success. Survival of human oocytes cryopreserved using current methodology is similar to that achieved with early-cleavage-stage embryos. Although evidence suggests that developmental potential may be compromised when current oocyte cryopreservation protocols are applied, there is a paucity of rigorously controlled studies in the literature. PMID- 17425823 TI - Co-culture of pre-antral ovarian follicles does not increase the yield of in vitro-matured oocytes in the mouse. AB - In-vitro culture of ovarian follicles offers the possibility of checking whether there are direct interactions between follicles. Gross follicle morphology and oocyte maturation were investigated as endpoints after co-culture of pre-antral mouse ovarian follicles. Pre-antral mouse follicles with a diameter of 95-110 microm (small) or >110-160 microm (large) were cultured. Pairs of different size or large like-sized follicles were cultured either in physical contact or without in 20 microl culture medium droplets under oil for 10 days. Individual culture of small and large follicles served as controls. On day 10 human chorionic gonadotrophin was administered. On day 11, 'in-vitro ovulated' cumulus-oocyte complexes where the cumuli oophori had expanded (termed 'mucified') were collected. The percentage of metaphase II (MII) oocytes was scored as primary endpoint. There were temporary differences between the degree of follicle attachment and diffusion of granulosa cells, but with the observations of antrum formation by day 10, there was no difference between different types of co cultures and individual culture of large follicles. The MII-to-follicle rate was highest for individual culture of large follicles (54.7%) followed by coculture of large like-sized follicles growing either in contact or not (40.1% and 42.6%), co-culture of different-sized follicles growing in contact or not (28.5% and 29.8%) and small follicles cultured individually (8.1%). In conclusion, mature oocyte yield was not increased by co-culture of mouse pre-antral ovarian follicles. PMID- 17425825 TI - Effect of aspiration vacuum on the developmental competence of immature human oocytes retrieved using a 20-gauge needle. AB - In-vitro maturation (IVM) of immature oocytes has been proposed as a potential alternative to conventional IVF treatment following ovarian stimulation. However, the effects of the oocyte retrieval conditions on subsequent development have not been well understood. This study assessed the effects of different aspiration vacuums during oocyte retrieval on the developmental competence of immature oocytes following IVM, IVF and embryo transfer, retrospectively. Immature oocytes were aspirated with 20-gauge needles with a vacuum of 180 or 300 mmHg. Immature oocytes were cultured in IVM medium for 26 h. All mature oocytes were inseminated by intracytoplasmic sperm injection (ICSI). Embryo transfer was carried out 2 or 3 days after ICSI. The percentage of cumulus-cell enclosed oocytes and of transferable embryos per retrieved oocytes in 180 mmHg (69.7% and 23.8%, respectively) were significantly higher (P < 0.01) than those in 300 mmHg (46.2% and 12.8%, respectively). The ongoing pregnancy rate per retrieval cycle in 180 mmHg (30%) was higher (P < 0.01) than that in 300 mmHg (4.3%). The data indicate that lower pressure of vacuum aspiration with a 20-gauge needle improves the developmental competence of immature oocytes following IVM, IVF and embryo transfer. PMID- 17425826 TI - Mullerian inhibiting substance is an accurate marker of ovarian response in women of advanced reproductive age undergoing IVF. AB - While multiple investigators have demonstrated that Mullerian inhibiting substance (MIS) concentration is an accurate marker of ovarian reserve, all previous reports have focused on patients aged 36 and younger. It remains to be seen if MIS concentration is a useful marker in patients with the highest prevalence of diminished ovarian reserve: those aged 37 and over. Day 2 MIS concentrations in patients aged 37 and over undergoing IVF were examined, and the predictive value of MIS concentration was compared to that of FSH and oestradiol concentration. Three groups of patients were studied: (i) patients who experienced cycle cancellation; (ii) patients who underwent oocyte retrieval; (iii) patients who were precluded from IVF due to elevated FSH concentrations. While FSH and oestradiol concentrations were statistically similar in groups 1 and 2, MIS concentrations were significantly lower in group 1. In group 2, MIS concentration positively correlated with the number of oocytes retrieved. In group 3, 91% had low or undetectable MIS concentrations. The results indicate that MIS concentration accurately predicts cycle cancellation that is belied by normal FSH and oestradiol measurements, and has a better correlation with the number of oocytes retrieved than FSH and oestradiol measurements. PMID- 17425827 TI - Cumulative live birth rates following IVF in 41- to 43-year-old women presenting with favourable ovarian reserve characteristics. AB - For women aged 41-43 years old, success rates in IVF are generally poor. This study aimed to assess cumulative live birth rate related to treatment costs over a maximum of three IVF cycles in selected women who were considered to still have adequate ovarian reserve. Fifty-five patients (38% of the total cohort, n = 144) were excluded from IVF treatment based on low antral follicle count (<5 follicles) and/or elevated basal FSH (>15 IU/l). Of those admitted, 66 (74%) actually started and completed a total of 125 IVF/intracytoplasmic sperm injection cycles. Treatment resulted in 10 live births (8% per cycle). Kaplan Meier survival analysis revealed a realistic cumulative live birth rate after three cycles of 17%. The direct medical costs per live birth were calculated to be approximately 44,000 euro. These results show that selection towards favourable ovarian reserve status in the female age group 41-43 years yielded disappointing results in terms of cumulative live birth rates after IVF. In view of the costs raised per live birth, improvement of selection parameters for treatment in this age group is warranted. PMID- 17425829 TI - High survival and hatching rates following vitrification of embryos at blastocyst stage: a bovine model study. AB - Cryopreservation of embryos at the blastocyst stage may provide an effective method to increase the cumulative pregnancy rate for each treatment cycle of ovarian-stimulated IVF. The objective of this study was to evaluate the survival rate and hatching rate of bovine blastocysts following vitrification using a method designed for oocytes, with a view to introducing this methodology into human assisted reproduction technology and reproductive medicine. Bovine blastocysts were produced from abattoir materials subjected to in-vitro maturation and in-vitro fertilization. Survival rate of the bovine blastocysts was 100% (94/94) following vitrification using a method designed for oocyte cryopreservation. There was no difference in the hatching rate of the bovine blastocysts between control (62.5%: 60/96) and vitrified (61.7%: 58/94) groups. The number of dead cells in the blastocysts was not significantly different between control (5.0 +/- 2.9) and vitrified (9.5 +/- 4.0) groups. In conclusion, the results of this study indicate that bovine blastocysts can be vitrified successfully using a procedure designed for oocyte cryopreservation. It is possible that this method may also be successful for the cryopreservation of human embryos. A further study into this is currently being organized. PMID- 17425830 TI - Prospective randomized comparison of Wallace and Labotect embryo transfer catheters. AB - The aim of this study was to compare the clinical performance of Wallace and Labotect embryo transfer catheters in a single assisted reproduction centre. A total of 260 women undergoing embryo transfer were randomized between Wallace and Labotect catheters. When both catheters failed to negotiate the cervical canal, the transfer was accomplished with a stiff catheter. Intention to treat analysis revealed that Wallace and Labotect embryo transfer catheters yielded statistically similar clinical pregnancy (44.6 versus 34.6%), implantation (23.2 versus 18.9%) and ongoing pregnancy (38.5 versus 27.7%) rates. As treated analysis revealed clinical pregnancy rates of 42.5 versus 35.4%, implantation rates of 22.3 versus 20.6% and ongoing pregnancy rates of 36.8 versus 28.3% with Wallace and Labotect catheters respectively. Catheter change due to unsuccessful negotiation of the internal cervical os was significantly more frequently necessary when embryo transfer was first intended with the Wallace than the Labotect catheter (P < 0.001; 33 and 2% respectively). Although not being statistically significant, the observed differences may be regarded as clinically important and may reach statistical significance in larger trials. More trials are necessary before reaching a definitive conclusion regarding the performance of the Labotect embryo transfer catheter. PMID- 17425831 TI - Proteomic profiling of forskolin-induced differentiated BeWo cells: an in-vitro model of cytotrophoblast differentiation. AB - Placental trophoblastic differentiation is characterized by the fusion of monolayer cytotrophoblasts into syncytiotrophoblasts. During this process of differentiation, several morphological and biochemical changes are known to occur, and this model has been employed to investigate the changes that occur at the gene and protein level during differentiation. Using the sensitive technique of proteomics [two-dimensional gel electrophoresis (2DGE)], changes in protein profile were evaluated in the control and forskolin-induced differentiated cells of trophoblastic choriocarcinoma BeWo cell line. Several proteins were differentially expressed in control and differentiated cells. Four major proteins were up-regulated as assessed by silver staining, and were further characterized as c-h-ras p 21 (phosphorylated), retinoblastoma susceptibility protein 1 and integrase interactor protein 1. These proteins are known to play an important role in growth arrest of cells, and thus may play a role in initiating the process of differentiation. PMID- 17425832 TI - Fertility assessment in non-infertile HIV-infected women and their partners. AB - The objective of the study was to assess the fertility of non-infertile couples seeking pregnancy in whom the woman was HIV infected. Therefore, a cross sectional study was conducted between January 1998 and March 2005. A standardized fertility assessment was performed in all the included couples. A total of 130 women and 121 men were evaluated. Their median age was 34 years (range 22-43). Only 7.2% of the women were severely immunocompromised. The majority of women had regular cycles. Only one woman had an active sexually transmitted disease at the time of evaluation. A tubal occlusion on hysterosalpingogram was present in 27.8% of the women with no proven fertility. In 50.5% of the women, hepatitis C virus co-infection was present. One-third of the male partners (38/121) was infected with HIV. Abnormal semen parameters were observed in 83.4% of HIV-infected and 41.7% of HIV-uninfected partners (OR = 7; 95% CI = 2.1-23). It is concluded that the great majority of the HIV-infected women seeking pregnancy had a good infection status. Because in many of the couples, the women presented unexplained tubal occlusions and the men presented semen alterations, a hysterosalpingography and semen analysis should be part of the preconceptional investigations. PMID- 17425833 TI - Use of three-dimensional ultrasound in evaluating the intrauterine position of a levonorgestrel-releasing intrauterine system. AB - This paper reports the first case of three-dimensional (3-D) transvaginal ultrasonography (TVS) imaging of malpositioned levonorgestrel-releasing intrauterine system (LNG-IUS). In patients carrying LNG-IUS, the application of 3 D TVS with the adjunctive volume contrast imaging in the coronal plane (VCI-C) and inversion rendering modes clearly display the correct spatial position of the LNG-IUS in relation to the uterine cavity much better than two-dimensional ultrasound. PMID- 17425834 TI - Obtaining metaphase spreads from single blastomeres for PGD of chromosomal rearrangements. AB - It has previously been shown that it is possible to obtain metaphase chromosomes from single blastomeres converted into metaphase in the cytoplasm of a mouse zygote. This method is highly labour intensive and cannot be performed outside the preimplantation genetic diagnosis (PGD) laboratory, so to overcome these limitations, a method was developed for obtaining metaphase spreads from single biopsied blastomeres using different chemicals. The substances tested were calyculin A, caffeine, paclitaxel and colcemid in a total of 496 disaggregated and 234 biopsied blastomeres from day 3 embryos. It was demonstrated that the optimal method involved a combined use of 'selective biopsy' (selection of the biopsied blastomere according to morphological criteria) and exposure to caffeine. This resulted in shortening the mean incubation time of biopsied blastomeres, with a metaphase formation rate of 80%. The method is simple for obtaining metaphases from single blastomeres, and may be implemented in clinical practice of PGD for structural rearrangements. PMID- 17425836 TI - Duplication of the sperm genome by human androgenetic embryo production: towards testing the paternal genome prior to fertilization. AB - There is currently no technique for evaluating the sperm genome before fertilization. However, sperm genome duplication could offer a way forward, whereby one of the sister blastomeres of a 2-cell haploid androgenetic embryo could be analysed. A method was developed for production of human androgenotes by enucleation of oocytes at telophase II (TII) after intracellular sperm injection (ICSI). The results were compared with those obtained via the more usual procedure of oocyte enucleation at metaphase II (MII) prior to ICSI. TII enucleation led to an improvement in the rate of embryo survival, increased the production rate of 1PN-embryos, and also the production of 2- to 8-cell-stage embryos (85.0, 74.9 and 65.8% in TII enucleation, versus 73.8, 48.9 and 33.3% in MII enucleation). Fluorescence in-situ hybridization (FISH) analysis of 30 2- to 5-cell androgenic embryos for two to seven chromosomes revealed the correct chromosome distribution in 76.7% of haploid human androgenotes. PMID- 17425837 TI - Preimplantation genetic diagnosis for fragile X syndrome using multiplex nested PCR. AB - Fragile X syndrome is caused by a dynamic mutation in the FMR1 gene. Normal individuals have <55 CGG repeats in the 5 untranslated region, premutation carriers have 55-200 repeats and a full mutation has >200 repeats. Female carriers are at risk of having affected offspring. A multiplex nested polymerase chain reaction protocol is described for preimplantation genetic diagnosis (PGD) of fragile X syndrome with simultaneous amplification of the CGG-repeat region, the Sry gene and several flanking polymorphic markers. The amplification efficiency was > or =96% for all loci. The allele dropout rate in heterozygotic females was 9% for the FMR1 CGG-repeat region and 5-10% for the polymorphic markers. Amplification failure for Sry occurred in 5% of single leukocytes isolated from males. PGD was performed in six patients who underwent 15 cycles. Results were confirmed in all cases by amniocentesis or chorionic villous sampling. Five clinical pregnancies were obtained (31% per cycle), four of which resulted in a normal delivery and one miscarried. This technique is associated with high efficiency and accuracy and may be used in carriers of full mutations and unstable high-order premutations. PMID- 17425838 TI - The ESHRE/ASRM consensus on polycystic ovary syndrome (PCOS)--an extended critical analysis. AB - The 'ESHRE/ASRM consensus on diagnosis, nomenclature and long-term health risks of polycystic ovarian syndrome (PCOS)' (conference in Rotterdam, Netherlands, March, 2003), which was criticized shortly after its oral presentation in 2003, is still being debated following its publication in 2004. Therefore, an extended, intensive analysis of all parameters and their combinations has been performed, differentiating between inclusion and exclusion criteria. By doing this, the profound significance of the endocrinological part of the syndrome is highlighted. The nomenclature, semantic meaning of terms, definitions, structural balancing, specificity, practicability, flexibility, limitations and reproducibility for both individualization, as well as grouping from the clinical and scientific perspective, have been examined in detail. This analysis supports previous critical comments, and reaches the conclusion that the recommendations of the consensus lack accuracy and specificity, a weakness that results in the risk of overdiagnosis and in a lack of reproducibility. Altogether, they fail to fulfil the criteria of universal guidelines on several levels. Instead, a profound paradigm shift is demanded, from an inconsistent perception to a more systematic, balanced and comprehensive view. A short indication on an alternative sweeping concept of a strictly defined, therapy-targeted classification and diagnostic procedure of female functional androgenization is presented. PMID- 17425840 TI - How fashions change in gynaecology and infertility. AB - In medicine the changes in therapies are the evidence of the progress in knowledge. But many examples concerning the treatment of early spontaneous abortion, late abortion, menopause, contraception and infertility show that gynaecologists and obstetricians change their treatments just as fashion designers change their styles. My 'Corner' on this occasion is an appeal for caution and humility. PMID- 17425841 TI - Chorionic gonadotrophin, genes and embryonic signals regulating the implantation window. PMID- 17425842 TI - [Vasculogenic mimicry in human primary gallbladder carcinoma and clinical significance thereof]. AB - OBJECTIVE: To investigate if there is vasculogenic mimicry (VM) in human primary gallbladder carcinoma and clinical significance thereof. METHODS: Seventy-four specimens of primary gallbladder carcinoma obtained from operation underwent HE staining and double staining of CD(31) and PAS to observe the existence of VM. The correlation of pathological examination and clinical outcomes was analyzed. RESULTS: VM was seen in 10 of the 74 (13.5%) specimens. VM was not correlated with age, sex, location, diameter, differentiation degree, Nevin stage, and invasion depth of tumor, and existence of lymph node metastasis; but was associated with histological type (chi(2) = 10.241, P = 0.017), hepatic metastasis (chi(2) = 11.904, P = 0.01), and poor overall survival (chi(2) = 5.771, P = 0.016). Cox analysis showed that existence of VM, invasion depth, lymph node metastasis, hepatic metastasis, and operational method were independent risk factors of the prognosis of primary gallbladder carcinoma. CONCLUSION: VM exists in human primary gallbladder carcinoma. Those cases of human primary gallbladder carcinoma with VM have a poorer prognosis. PMID- 17425843 TI - [Expression of Fas, Fas ligand, Fas-associated death domain protein, caspase 8 and mutant P53 protein in esophageal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the expression of five apoptosis-related proteins, Fas, Fas ligand (FasL), Fas-associated death domain protein (FADD), caspase 8, and mutant p53, in human esophageal squamous cell carcinoma (ESCC) tissue, and analyze the association of these proteins with ESCC malignant progression. METHODS: 116 ESCC specimens obtained during operation. Tissue microarray composed of the 116 specimens of cancerous tissues and corresponding paracancerous normal epithelium tissues was constructed. The expression of Fas, FasL, FADD, caspase 8, and mutant p53 was detected in the ESCC tissues and paracancerous normal epithelium tissues and analysis was conducted for the correlation between the expression of these proteins and the pathoclinical features and prognosis. involvement, differentiated grade, pTNM stages and disease-free survival. RESULTS: The positive rate of Fas in the ESCC tissues was 41.4%, significantly lower than that in the normal squamous epithelium was 95.7%, P < 0.001). The positive rates of FasL and FADD in the ESCC tissues were 76.7% and 50.0%, both significantly higher than those in the normal squamous epithelium (39.7% and 7.8%, both P < 0.001). Caspase 8 was strongly positive in the whole normal esophageal epithelium tissue except basal and superbasal cells, but negative in ESCC. Mutant p53 protein was negative in the normal esophageal epithelium tissue, with only several cases positive in the basal cells, but was diffusely positive in ESCC tissues with a positive rate of 37.1%. The expression of Fas in the well and moderately differentiated ESCC tissues was significantly higher than in the poorly differentiated ones (P = 0.022). The patients with positive expression of FADD had lower disease-free survival rate (P = 0.0028). The expression of Fas, FasL, caspase 8, and mutant p53 was not related with disease-free survival rate (P > 0.05). CONCLUSION: The apoptosis-related markers, such as Fas, FasL, FADD, caspase 8, and mutant p53 protein may play important roles in the development and progression of ESCC, and FADD can be used as a marker to predict the advance and prognosis of ESCC. PMID- 17425844 TI - [Study of the correlation between HER-2 gene and lymphangiogenesis and their prognostic significance in human breast cancer]. AB - OBJECTIVE: To demonstrate the molecular mechanism of poor prognosis in HER-2 overexpressing breast carcinoma, to study the correlation between angiogenesis/lymphangiogenesis and HER-2, and the effect of them in the progression and prognosis, so as to provide novel molecular markers for lymph node metastasis and prognosis in breast carcinoma. METHODS: (1) By using TaqMan real-time quantitative reverse transcription polymerase chain reaction (real-time qRT-PCR) technique and immunohistochemistry (IHC), the mRNA and protein expression of VEGF-C were detected in SKBR-3 and MDA-MB-231 cell lines before and after the interference with Herceptin. (2) Real-time qRT-PCR was performed to detect the mRNA expression of VEGF-C and -D genes in 67 cases of primary breast infiltrating ductal carcinoma. (3) 160 formalin-fixed paraffin-embedded infiltrating ductal carcinomas were selected. The protein expression of HER-2, VEGF-C, lymphatic vessel density (LVD) and lymphatic vessel invasion (LVI) were detected by IHC. RESULT: (1) After interference with Herceptin, the VEGF-C mRNA and protein expression of sKBR-3 the cells decreased significantly. In primary breast cancer, the protein expression of HER-2 was positively correlated with VEGF-C protein expression (r = 0.215, P < 0.05). There was a significantly positive correlation between VEGF-C protein expression and LVD (r = 0.248, P < 0.01). (2) VEGF-C mRNA expression in the patients with lymph node metastasis was higher than that in the patients without lymph node metastasis (1.6 +/- 0.5 vs 1.1 +/- 0.5) (t = 2.196, P < 0.05). Logistic regression model showed HER-2, LVD and LVI were important factors influencing lymph node metastasis (all P < 0.05). (3) Kaplan-Meier analysis indicated that HER-2 protein overexpression, VEGF-C protein overexpression, and high LVD and LVI were associated with patient's worse disease-free survival (P(uni) < 0.05). Cox regression analysis indicated that HER 2 and LVI were the independent prognostic factors of breast cancer (both P(mult) < 0.05). CONCLUSION: Through up-regulating VEGF-C expression, HER-2 overexpression induces lymphangiogenesis, promotes metastasis, and results in the poor prognosis of breast cancer. VEGF-C expression, LVD and LVI are novel prognostic indicators of breast cancer. PMID- 17425845 TI - [Clinicopathological study on nodal molecular staging of non-small-cell lung cancer]. AB - OBJECTIVE: To investigate the practicability of detecting the micrometastasis in lymph nodes of no-small-cell lung cancer (NSCLC) by means of reverse transcriptase-polymerase chain reaction (RT-PCR). METHODS: Regional lymph node samples were collected during operation from 25 patients with NSCLC randomly selected. The lymph node sample from each patient was divided into 2 groups: lymph nodes of hilum of lung and of mediastinum. Every lymph node was divided into two parts of the same size. One half part of the lymph node was examined by hematoxylin eosin (H&E) staining. If HE staining discovered metastasis, further examination was not needed. If HE staining failed to discover metastasis, then all the remaining lymph node samples of each patient were mixed together to undergo RT-PCR for cytokeratin 19 (CK(19)), a tissue marker of epithelium and epithelial tumors. RESULTS: (1) 195 lymph nodes from 25 patients with NSCLC were examined by H&E staining. 30 lymph nodes in 9 patients showed gross nodal metastasis and none showed micrometastatic tumor cells. (2) Of the 39 groups of mixed regional lymph node samples which were diagnosed to be devoid of metastases by H&E staining, 11 groups were found to have positive reactions to CK(19) mRNA. (3) Six of the sixteen patients staged as PN(0) had hilum lymph nodal micrometastasis, while 5 of nine patients with stage PN(1) had mediastinal lymph nodal micrometastasis (chi(2) = 54.063, P = 0.0043). CONCLUSION: (1) H&E staining can accurately detect gross nodal metastasis in the lymph nodes of the patients with NSCLC, but is unfit for detecting lymph nodal micrometastasis. (2) RT-PCR can facilitate the detection of occult micrometastatic tumor cells in the lymph nodes of NSCLC, and its assessment of nodal micrometastasis can provide a refinement of molecular stage for partial patients with stage I to II. PMID- 17425846 TI - [Microsurgical anatomy and surgical approach of the ambient cistern]. AB - OBJECTIVE: to describe detailed anatomy and compare four surgical approach of the ambient cistern, to assist in the process of conceptualizing a reliable surgical approach to allow for a successful course of surgery. METHODS: The microsurgical anatomy of the ambient cistern and four surgical approach was studied in 12 formalin-fixed brain specimens (24 hemispheres). Four simply formalin-fixed, the cerebral arteries of eight brain specimens were perfused with colored silicone, and they were dissected with the aid of an operating microscope. Special attention was paid to the anatomic structures limiting visualization in each approach. RESULTS: The ambient cistern extends from the posterior margin of the crural cistern to the lateral edge of the midbrain colliculi, and round the lateral surface of the upper portion of the brainstem; The ambient cistern mainly contained P(2) segment of the posterior cerebral artery, superior cerebellar artery, anterior choroidal artery, basal vein and trochlear nerve. The subtemporal approach allow for excellent exposure of the lower half of the ambient cistern, the vein of Labbe and the parahippocampal gyrus obstacle to exposure through this approach; The transtemporal transchoroidal approaches exposed the upper half of the ambient cistern of non-dominance hemisphere, its major drawback is the need to perform a corticectomy in the temporal lobe. In addition, the vein of Labbe extended far enough anteriorly to limit the cortical incision in the temporal lobe. The transinsular transchoroidal exposed the anterior upper half of the ambient cistern. Its major disadvantage is damage the temporal stem, the longer the inferior limiting sulcus, the shorter the incision, the better for preservation the temporal stems; The occipital transtentorial approaches exposed the posterior lower half of the ambient cistern, the parahippocampal gyrus also blocked to exposure through this approach. CONCLUSION: Surgical approaches to lesions of the ambient cistern must be tailored to the site of the pathological findings. Position of the vein of Labbe, and the midpoint of the rounded medial edge of the parahippocampal gyrus, is key to determine Surgical Approach. PMID- 17425847 TI - [Assessment of preclinical cognition impairment in hypertension patients with functional magnetic resonance imaging]. AB - OBJECTIVE: To explore whether the hypertension patients with no clinical cognitive impairment manifestations have certain brain dysfunctions. METHODS: Twenty-five moderately to severely hypertensive patients, males and females, aged 63.0 +/- 1.6 (60 approximately 65), with a disease history if 5 to 10 years, and 25 sex, age, and educational level-matched healthy persons underwent tests by mini-mental status examination (MMSE) scales, state anxiety inventory (STAI-S) and trait anxiety inventory (STAI-T), and then underwent two functional magnetic resonance imaging (fMRI) studies. In Experiment 1, the subjects were demanded to listen actively to the meaningless words (pseudowords) and in Experiment 2 the subjects listened actively to real words and make the valence (abstract or concrete) of the words in silence. The subjects were told to listen passively the noise from the MR scanners during the resting period, which was used as the control task. The fMRI data were analyzed with statistical parametric mapping (SPM99) software. RESULTS: The MMSE score of the patient group was 29.3 +/- 1.1, not significantly different from that of the control group (29.6 +/- 0.5, P > 0.05). The STAI-S score of the patient group was 47 +/- 5.3748, significantly higher than that of the control group (41.6 +/- 4.9777, P < 0.05). The STAI-S score of the patient group was 45 +/- 3, not significantly different from that of the control group (43 +/- 4, t = 1.0619, P = 0.3032). In Experiment 1, the activated patterns and deactivated patterns were nearly similar for the patient and control groups. The activated regions included the bilateral superior temporal lobe, bilateral inferior frontal cortex and supplementary motor areas. In Experiment 2, the activated patterns were also nearly similar for these 2 groups. The regions included the bilateral superior temporal lobe, bilateral inferior frontal cortex, left angular gyrus, bilateral superior frontal cortex, bilateral cerebellum, premotor areas, and supplementary motor areas. The deactivation patterns were similar for the patients and controls both in Experiment 1 and Experiment 2. These regions were medial prefrontal cortex, posterior cingulate and bilateral inferior parietal cortex, which nearly overlapping with the default model network proposed by Raichle et al. However, the activation and deactivation magnitude and extent of the brain were significantly greater for the patients than for the controls both in Experiment 1 and Experiment 2. CONCLUSION: Some brain dysfunction may have already existed in patients with moderately to severely hypertension disorders, though their behavior performance scores are within the normal range of ages. The fMRI technique can be a useful tool to detect the preclinical brain abnormalities. PMID- 17425848 TI - [Effects of peroxisome proliferator-activated receptor gamma and its ligands on cytotrophoblast invasion in first trimester of pregnancy and mechanism thereof]. AB - OBJECTIVE: To investigate effects of peroxisome proliferator-activated receptor gamma (PPARgamma) and its ligands on cytotrophoblast invasion and its influence on matrix metalloproteinase (MMP)-2 and MMP-9 activities on cytotrophoblast cells. METHODS: Samples of fresh placental trophoblast tissue were obtained from the pregnant women in first trimester. Cytotrophoblasts were isolated, cultured, and added with PPARgamma. Immunocytochemistry and inverted microscopy were used to examine the protein expression of PPARgamma in the cytotrophoblasts. Isolated cytotrophoblasts were inoculated in the Transwell chamber The natural stimulatory ligand of PPARgamma15-deoxy-delta (12, 14)-prostaglandin J2 (15d-PGJ2) and synthesized stimulatory ligand of PPARgamma troglitazone (TGZ) of the concentrations of 0, 1, and 10 micromol/L respectively were added respectively to examine the invasion ability of the cytotrophoblasts. Immunofluorochemistry and confocal technique and RT-PCR were used to examine the expression of MMP-2 and MMP-9. RESULTS: PPARgamma protein expression was detectable in the cytotrophoblasts, mainly in the nuclei. The invasion indexes of the cytotrophoblasts stimulated by 15d-PGJ2 and TGZ of different concentrations were significant lowered in a concentration-dependent manner, the effect of 15d-PGJ2 being stronger than that of TGZ (P < 0.05). After stimulated by 15d-PGJ2 and TGZ of different concentrations the mRNA expression and protein expression of MMP-9 and MMP-2 in the cytotrophoblasts were all inhibited significantly (all P < 0.05). CONCLUSION: PPARgamma plays an important role in the modulation of trophoblast invasion, and PPARgamma ligands can inhibit the trophoblast invasion through downregulating MMP-2 and MMP-9 expressions. PMID- 17425849 TI - [Expression of cyclin B1 and cyclin-dependent kanasel in ectopic and eutopic endometrium tissues of patients with endometriosis]. AB - OBJECTIVE: To investigate the possible roles of cyclin B1 and cyclin-dependent kanase1 cyclin-dependent kanase1 play in the pathogenesis and development of endometriosis and their association with the ovarian hormones. METHODS: Twenty nine specimens of ectopic endometrium tissues and 20 specimens of eutopic endometrium tissues were obtained from 29 patients with endometriosis, aged 24 approximately 46. Thirty specimens of endometrium form 30 women without endometriosis were used as controls. The intracellular location of cyclin B1 and Cdc2 was detected by microscopy. Western blotting, RT-PCR, and immunohistochemistry were employed to examine the mRNA expression and protein expression of cyclin B1 and Cdc2. Serum estrogen and progestogen were detected. RESULTS: The expression level of cyclin B1 in the ectopic endometrium from the women with endometriosis was significantly higher than that in the ectopic endometrium tissues from the women with and without endometriosis (both P < 0.05). No significant difference was found between the expression level of cyclin B1 in the ectopic endometrium tissues from the women with and without endometriosis (both P > 0.05). The Cdc2 expression levels were not significantly different among th3 3 groups and the proliferative and secretary stage of endometrium (all P > 0.05). Cyclin B1 expression level was positively correlated with the serum estrogen level, and negatively correlated with the serum progestogen level, and Cdc2 expression was not correlated with the serum sex hormone levels. CONCLUSION: The expression of cyclin B1 in the ectopic endometrium is higher than normal. Cyclin B1 may be involved in the proliferation of endometrium in endometriosis. PMID- 17425850 TI - [Treatment of stroke in rats with bone marrow mesenchymal stem cells]. AB - OBJECTIVE: To investigate the effects of treatment of stroke in rats with bone marrow mesenchymal stem cells (BMSCs) and mechanism thereof. METHODS: Bone marrow of a healthy volunteer was collected and the BMSCs were separated with density gradient centrifugation. The hBMSC were cultivated and harvested until the third passage. A number of adult male Sprague-Dawley rats received corresponding behavioral training before surgery and underwent transient middle cerebral arterial occlusion (MCAO) for 2 hours. Sixty of them showing the scores of 6 approximately 12 according to the modified neurological severity score system were randomly divided into 2 groups: treatment group (n = 48, injected into the cortex around the ischemic areas with hBMSCs 3x10(5)/15 microl) and control group (n = 12, injected with D-Hanks solution 15 microl 24 hours after the establishment of MCAO models. Morris water maze test, Rotarod test and adhesive removal test were performed since the 4th day to the 32 day after transplantation once every 3 days. 1, 2, 3, and 4 weeks after the transplantation 12 rats from each group were killed randomly to take out their brains. Immunofluorescence was used to identify the migration, survival and differentiation of the hBMSC. RESULTS: A large number of hBMSC could be seen within 2 weeks after transplantation. The number of hBMSC decreased since the 21st day after transplantation and few cells could be found at the end of 1 month after. No definite evidence supported the differentiation of neural cells derived from the hBMSCs during the whole process. Morris water maze test showed that the mean escape time 1 week after transplantation of the treatment group was (69 +/- 10) s, significantly shorter than that of the control group [(120 +/- 0) s, P < 0.05] The significant difference persisted until the 4(th) week (P > 0.05). Rotarod test with the speed of 10 r/min showed that the mean latency period 10 days after transplantation of the treatment group was (167 +/- 18) s, significantly longer than that of the control group [(37 +/- 19) s, P < 0.05]. The significant difference persisted until the experimental terminal. The adhesive-removal test showed that the mean latency period 13 days after transplantation of the treatment group was (33 +/- 8) s, significant shorter than that of the control group [(84 +/- 13) s, P < 0.05]. The significant difference persisted until the experimental terminal. CONCLUSION: Injection of hBMSCs into brain cortex improves neurological functional recovery after stroke. The transplanted cells can migrate and survive for a certain period, but no hBMSC express proteins phenotype of neural cells. PMID- 17425851 TI - [Osteoblastic differentiation and in vivo osteogenic activity of marrow-derived mesenchymal stem cells stimulated by Tacrolimus: experiment with rats]. AB - OBJECTIVE: To investigate the effect of the immunosuppressant Tacrolimus (FK506) on the osteoblastic differentiation and in vivo osteogenic inducement of bone marrow-derived mesenchymal stem cells (MSCs). METHODS: MSCs were derived from Fischer 344 rats. Some MSCs were cultured with L-ascorbic acid-2-phosphate (AsAP) and beta-glycerophosphate (beta-GP) or FK506 plus AsAP and beta-GP. The alkaline phosphatase (APase) activity and calcium deposition were detected 4, 8, 12, and 16 days after the culture. Scanning electron microscopy was used to examine the calcified nodules. Northern blotting was used to detect the mRNA expression of osteocalcium. Multiparous beta-tricalcium phosphate (beta-TCP) ceramic cubes were dipped into 2 kinds of suspension of MSCs, treated by FK506 + AsAP + beta-GP or AsAP + beta-GP, so as to produce 48 pieces of MSCs/beta-TCP complex that were randomly divided into 2 equal groups to be cultured with AsAP + beta-GP or AsAP + beta-GP + FK506 for 4 weeks. The these pieces were transplanted into the subcutaneous sites of the rats' backs and were taken out 4 and 8 weeks later respectively for histological examination. RESULTS: In vitro assays showed that the APase activity, calcium deposition, expression of osteocalcin mRNA of the FK506 + AsAP + beta-GP group at any time points were all significantly higher than those of the AsAP + beta-GP group (all P < 0.05). SEM showed that since the 16th day after culture calcified nodules began to be seen in the FK506 + AsAP + beta-GP group. Since the 4th weeks after transplantation remarkable new bone formation could be seen in the FK506 treated MSCs/beta-TCP complexes in comparison with those MSCs/beta-TCP complexes without treatment with FK506. CONCLUSION: Greatly enhancing the in vitro osteoblastic differentiation and in vivo osteogenesis of MSCs, FK506 has a potential value as a bone growth factor and may improve the clinical result of bone transplantation used to treat large bone defect. The results of this experiment also contributes to a better understanding on the mechanism of immunosuppressants. PMID- 17425852 TI - [Effects of antisense oligonucleotide to nuclear factor-kappaB on the development of bleomycin-induced pulmonary fibrosis and IL-4 expression therein: experiment with mice]. AB - OBJECTIVE: To investigate the expression of nuclear factor-kappaB (NF-kappaB) in the cells in the bronchoalveolar lavage fluid (BALF) of idiopathic pulmonary fibrosis (IPF) and its effects on IL-4 expression therein, and to investigate the therapeutic role of antisense oligonucleotide (ASON) to NF-kappaB on IPF. METHODS: C57BL/6 mice were randomly divided into 4 groups: bleomycin (BLM) group (n = 35, injected with BLM through caudal vein), control group [n = 20, injected with normal saline (NS) via caudal vein], ASON group (n = 35, injected with ASON to p65, a subunit of NF-kappaB, at the dose of 900 microg), and SON group (n = 35, injected with sense oligonucleotide to p65 subunit). Six hours after intravenous injection, the BLM, ASON, and SON groups were treated with BLM-A5 (5 mg/kg dissolved in 20 microl NS) by intratracheal installation, and the control group was treated with NS (20 microl). 0.5, 1, 3, 7, 14, and 28 days following intratracheal instillation of BLM or 0.5, 1, 14 days following intratracheal instillation of NS, 5 mice of every group were sacrificed and bronchoalveolar lavage was performed. The BALF was collected and assayed with ELISA for IL-4. Immunohistochemistry (IHC) and microscope image analysis were completed to detect the expression of p65 and IL-4 in the bronchoalveolar lavage cells. Another 5 mice from each group were sacrificed 28 days after intratracheal instillation with their total right lungs taken out to undergo pathohistological examination. The content of hydroxyproline in the left lung was detected by high performance liquid chromatography and ELISA. RESULTS: (1) Twenty-eight days after intratracheal instillation, the BLM and the SON groups showed consolidation of the lung parenchyma with loss of the alveolar architecture and increased cellularity, while the ASON and control groups showed no significant pulmonary consolidation or fibrosis. (2) Twenty-eight days after intratracheal instillation, the hydroxyproline content of the BLM group was 876.8 +/- 91.1 nmol/lung, significantly higher than that of the control group (347.6 +/- 53.9 nmol/lung, t = -9.833, P < 0.001); the hydroxyproline content of the ASON group was 505.6 +/- 34.8 nmol/lung, significantly lower than that of the BLM group (t = -9.862, P < 0.001); however, the hydroxyproline content of the SON group was 775.2 +/- 68.9 nmol/lung, not significantly different from that of the BLM group (t = 2.118, P = 0.102). (3) One day after the intratracheal instillation of BLM, the value of average integral optical density of p65 in the bronchoalveolar lavage cells of the BLM, SON, and ASON groups were 275 +/- 13, 233 +/- 60, 233 +/ 60, and 126 +/- 34 respectively, all significantly higher that of the control group (38 +/- 18, t = 27.350, 8.039, and 6.107, P < 0.001, = 0.001, and = 0.004), that of the ASON group being significantly lower than those of the BLM and SON groups (t = 7.664 and -3.407, P = 0.002 and 0.027). (4) IHC showed that 1 day after the intratracheal instillation, the value of average integral optical density of IL-4 of the BLM, SON, and ASON groups were 134 +/- 16, 128 +/- 2, and 80 +/- 9 respectively, all significantly higher than that of the control group (33 +/- 12, t = 10.346, -5.927, and 5.313, P < 0.001, = 0.004, = 0.006), that of the ASON group being significantly lower than those of the BLM and SON groups (t = 6.967 and -3.591, P = 0.002 and 0.023). (5) ELISA showed that 1 day after the intratracheal instillation the IL-4 level BLM, SON, and ASON groups were (20.8 +/ 7.2) ng/L, (21.4 +/- 8.0) ng/L, and (9.7 +/- 1.4) ng/L respectively, all significantly higher than that of the control group [(1.6 +/- 3.6) ng/L, t = 6.494, 4.143, and 4.331, P = 0.003, 0.014, and 0.012], that of the ASON group being significantly lower than those of the BLM and SON groups (t = -3.553 and 3.577, P = 0.024 and 0.023) (6) Correlation analysis showed that 1 day after intratracheal instillation the expression of p65 was positively correlated with IL-4 expression in the bronchoalveolar lavage cells in the treatment group (r = 0.890, P < 0.05) and the ASON group (r = 0.909, P < 0.05). CONCLUSION: The expression of NF-kappaB is significantly increased and augments the expression of IL-4 indirectly in the BALF cells during the process of BLM-induced lung fibrosis. ASON significantly inhibits the NF-kappaB activation and the IL-4 expression, and may be useful in gene therapy for pulmonary fibrosis. PMID- 17425853 TI - [Rotating three-dimensional dynamic culture of osteoblasts seeded on segmental scaffolds with controlled internal channel architectures for construction of segmental tissue engineered bone in vitro]. AB - OBJECTIVE: To explore the feasibility, effectiveness, and mechanism of culturing osteoblasts on calcium phosphate cement (CPC) scaffolds with controlled internal channel architectures in a rotating bioreactor, and to develop a novel method for construction of segmental tissue engineered bone in vitro. METHODS: Self hardening CPC scaffolds with controlled internal channel architectures were designed and fabricated using computer aided design (CAD) and indirect rapid prototyping (RP) techniques. A rotating bioreactor was developed. Osteoblasts were isolated from the skull of rabbit and seeded onto the CPC scaffolds, cultured for up to 21 days in static or rotating three-dimensional (3D) dynamic conditions. 7, 14, and 21 days after the incubation the proliferation, metabolic activity, and differentiation of the osteoblasts were determined by MTT, glucose consumption, and alkaline phosphate activity (ALP) assays respectively. The distribution of cells throughout the scaffolds was observed by scanning electron microscopy (SEM) and the sphere like structures which the SEM images showed within the extracellular matrix were assessed by energy dispersive X-ray (EDX) analysis. RESULTS: At all culture time points, the rotatingly cultured constructs demonstrated greater proliferation, metabolic activity, and osteoblastic differentiation than those of the statically cultured constructs as evidenced by MTT, glucose consumption and ALP assays. SEM indicated that 21 days after the distribution of cells in the scaffolds in the rotating culture was much more uniform than in the static culture. The sphere like structures was identified as calcium phosphate nodules by EDX analysis. CONCLUSION: As a novel method for construction of segmental tissue engineered bone in vitro, the rotating 3D dynamic culture of osteoblasts on CPC scaffolds with controlled internal channel architectures improves the properties such as proliferation, metabolic activity, osteoblastic differentiation, and uniform distribution of the seeded cells over those that maintain in static culture. PMID- 17425860 TI - [The study of molecular imaging should be strengthen]. PMID- 17425861 TI - [The infection and immunity of HIV: hope and thought]. PMID- 17425863 TI - [Committed differentiation of transplanted bone derived mesenchymal stem cells and their potential to amend damaged liver functions: in vivo experiment with mice]. AB - OBJECTIVE: To observe whether bone mesenchymal stem cells (MSCs) have the potential to transdifferentiate into functional hepatocyte-like cells in the special "niche" as well as the therapeutic feasibility to repair damaged liver in mice. METHODS: 20 nude mice were randomly divided into four groups (n = 5 in each group): Group A: 1.0 ml/kg of carbon tetrachloride (CCl(4)) (dissolved in olive oil by ratio of 1:1) was injected into the peritoneum of mice twice a week for 5 weeks. GFP-positive MSCs (1 x 10(6) cells) were injected into the caudal tail vein 1 week after the first dose of CCl(4); Group B: treated with CCl(4) as in A, but received the same volume of saline; Group C: normal nude mice with GFP positive MSCs Transplanted in the same way as in A. Group D: normal controls. 4 weeks after the cell transplantation, all animal subjects were killed. Liver function tests (LFT), histology of HE and Masson staining as well as double immunofluorescent staining for GFP and albumin were studied in all groups. RESULTS: The hepatic fibrosis in group A & B confirmed the success of model for liver damage and there was no marked difference in the percent of the area occupied by collagen between two groups (10.5 +/- 1.5 vs 12.7 +/- 1.6, t = 2.238, P > 0.05). GFP-positive MSCs were mainly observed around portal area or interspace of lobules in group A. Some of GFP-positive cells also express albumin (35% +/- 7%). While in group B, C or D, there is no such findings. The level of serum albumin in group A was higher than that in group B (24.4 g/L +/- 3.3 g/L vs 18.6 g/L +/- 2.9 g/L, P < 0.05) while the level of ALT was also different between two groups (121 U/L +/- 21 U/L vs 192 U/L +/- 29 U/L, P < 0.05). CONCLUSION: The stimulus of persistent liver damage might enhances the migration of MSCs to the liver, in which some of the MSCs have the potential to transdifferentiate into hepatocyte-like cells. Transplantation of MSCs might amend the damaged tissue of host liver to a certain extent. PMID- 17425864 TI - [MRI monitoring ultra-small superparamagnetic iron oxide (USPIO) particle labeling C6 rat glioma cells]. AB - OBJECTIVE: To monitor the effects of labeling C6 rat glioma cells with different concentrations of USPIO in vivo and in vitro. METHODS: C6 rat glioma cells of 1 x 10(6), 2 x 10(6) and 1 x 10(7) were labeled with 0 microg/ml, 25 microg/ml, 50 microg/ml USPIO, The signal intensity of cells were evaluated by MRI with T(1)WI, T(2)WI and GRE/30 degrees sequences in vitro. 1 x 10(6) of C6 glioma cells were labeled with 0 microg/ml, 25 microg/ml, 50 microg/ml USPIO and inoculated into the right frontal lobe of 2 rats under stereotaxis apparatus respectively (total 6 rats), Same MRI parameters were used just as above. Iron particle density and cells was measured by HE and Prussian blue stain under microscopy. RESULTS: Different cell population was cultured with 0 microg/ml, 25 microg/ml, 50 microg/ml USPIO about 12 hours. The MR signal intensity of labeling cells were inversely correlated with the different concentration of USPIO groups in T(2)W and GRE/30 degrees imaging (t = 4.19, 3.38, P < 0.05) in vitro. There was an inversely correlation between the labeling cell population and the signal intensity at the same concentration of USPIO (t = 5.16, 2.35, 4.41; P < 0.05). Dyeing degree of labeling cells stained by Prussian blue gradually deepened from 25 microg/ml to 50 microg/ml by microscopy. In vivo MRI can clearly show the cells labeled with 25 microg/ml USPIO. CONCLUSIONS: Iron particle density in the rat glioma cells were gradually increased with the concentration of USPIO. The MR signal intensity was inversely correlated with the cell population at the same condition. 25 microg/ml USPIO labeling rat glioma cells were enough for in vivo monitoring by MRI. PMID- 17425865 TI - [Characteristics and in vitro imaging study of matrix metalloproteinase-2 targeting activable cell-penetrating peptide]. AB - OBJECTIVE: To study invasively imaging MMP2-positive tumor cell by paramagnetic Gadolinium or fluorescein carried by a activable cell penetrating peptides. METHODS: To label Fluorescein-5-isothiocyanate (FITC) and gadopentetate with the activable cell-penetrating peptides by a solid-phase synthesis method. Identification by TOF mass spectrum (TOF-MS). Isoelectric point of the activable cell penetrating peptides is determined by disc electrophoresis. T1 relacion of gadopentetate labeled with the activable cell-penetrating peptides (B) in water were determined on 400 MHZ NMR. Human lung cancer cell lines: A549 were respectively stained by FITC labeled with ACPPs (A) or FITC alone. MMP2 expression and activity were determined by zymography. T1WI signal of A549 incubated with 120 nmol/ml B or Diethylenetriaminepentaacetic Acid-Gadolinium (Gd DTPA) for different times were obtained by 1.5T MRI. The location of B in A549 was detected by Transmission Electron Microscopy. Visualization analysis and half quantitative analysis were used to determine the signal characteristics. The ANOVA analysis and the paired samples t test were performed by SPSS 13.0. RESULTS: MALDI TOF-MS molecular weigh of A and B respectively is 3789.74 and 3911.93. Isoelectric point is 11.005T1 Relacion of 0.5 mmol/L Gd-DTPA and B at 17 degrees C respectively is (0.052 +/- 0.01) sec and (0.050 +/- 0.001) sec. Fluorescein uptake assays showed that A translocated into A549 but would be inhibited by MMP2 antibody. Zymography showed both active-MMP2 (67000) and pro MMP2 (72000) expressed byA549. MR imaging showed A549 incubating with B had a high T(1) signal, and the signal of A549 incubating with Gd-DTPA is similar with that of the control group. Furthermore, ANOVA suggested that the T(1) signal intensity of A549 incubating with B was effected by incubating time (F = 267.569, P < 0.001) and increasing in a time-dependent fashion at the observed time. There is no difference between the T(1) signal intensity of A549 incubating with Gd DTPA and the control group (P > 0.05). TEM showed A in cytoplasm and nucleus. CONCLUSION: The study in vitro suggests that the MMP-2 activable cell-penetrating peptides bearing contrast media can detect the MMP2-positive tumor cell. PMID- 17425867 TI - [Gadolinium-loaded nanoparticle as a novel molecular imaging contrast agent for magnetic resonance imaging]. AB - OBJECTIVE: To synthesize a novel gadolinium-loaded nanoparticle as a molecular imaging contrast agent for magnetic resonance imaging. METHODS: Gadolinium ion was incorporated within a silica nanoparticle. The size of this nanosized particle was determined by using transmission electron microscope. The spin-echo (SE) images of nine nanoparticle dilutions in vitro were obtained from a 1.5 T clinical scanner, compared with gadolinium diethylene triaminepenta acetate (Gd DTPA). In vivo distribution of nanoparticle in Balb/c nude mice and Balb/c nude mice with nasopharyngeal carcinoma (NPC) xenografted CNE-2 tumors was studied at MR imaging, 30 sec, 5 min, 30 min, 1 h, 2 h, 4 h, and 24 h after intravenous administration. RESULTS: The gadolinium-loaded nanoparticle was short rod-shaped, and approximately 30 to 40 nm in diameter. The value of longitudinal relaxativity (r(1)) of gadolinium nanoparticle was much higher than that of Gd-DTPA. Thirty minutes after injection the gadolinium nanoparticle, the signal intensities of liver, kidney and xenografted tumor increased from 226 +/- 10, 283 +/- 7 and 195 +/- 5 to 352 +/- 12, 328 +/- 10 and 245 +/- 7, respectively. The dynamic MRI scanning showed that gadolinium nanoparticles were mainly distributed in liver after intravenous administration. Strong enhancement was also detected in CNE-2 xenografted tumors. CONCLUSION: A new gadolinium-loaded nanoparticle with high relaxativity was synthesized successfully, and might serve as a carrier for magnetic resonance molecular imaging. PMID- 17425868 TI - [Clinical and gene research of X-linked lymphoproliferative disease in a Chinese family]. AB - OBJECTIVE: To verify the pathogenesis in Chinese and to investigate the genetic rule of X-linked lymphoproliferative disease (XLP) therein. METHODS: The case history of a proband of XLP, male, aged 1 year and 5 months, who died 40 days after hospitalization, was reviewed. Fourteen his family members were interviewed for the development history, anamnesis, and underwent physical examination. Single-strand conformation polymorphism (SSCP-PCR) and sequencing were used to detect the SH2D1A mutation among the elder sister, younger brother, and parents of the poband. RESULTS: The proband and his elder brother suffered with virus associated hemophagocytic syndrome and both died in 40 days after the disease coming on in the last two years in succession. The second exon of SH2D1A of the younger brother of the proband showed a nonsense mutation in SH2D1A gene: the C-T nucleotide substitution at nucleotide position 462 result in a stop codon and pre mature termination of protein synthesis. The mother was proved as mutation heterozygote of the C and T nucleotide on the same site. The other members of the family were proved normal. The clinical manifestation of the younger brother of the proband was Langerhans cell histiocytosis. CONCLUSION: Langerhans cell histiocytosis may be a new clinical phenotype of XLP. The gene of SH2D1A is responsible for the disease of XLP in Chinese too. The newly developed method of SH2D1A mutation analysis may be suitable in the diagnosis of XLP in Chinese. PMID- 17425869 TI - [Hereditary hemorrhagic telangiectasia caused by mutation in intron 4 of ALK1 gene: analysis of a HTT family]. AB - OBJECTIVE: To investigate the molecular pathogenesis of hereditary hemorrhagic telangiectasia (HHT). METHODS: Peripheral blood samples were collected from a HTT family, including the proband, female, aged 48, and her mother, elder brother, elder sister, younger brother, and son. HHT gene mutations were identified by PCR SSCP and DNA sequencing and confirmed by reverse sequencing. Ectopic transcripts of RT-PCR were used to confirm the characteristics of the mutation in non canonical splicing site (IV S4 + 3 a > t). RESULTS: A mutational segment of PCR product of exon 4, exon-intron boundaries and the 3', 5' untranslated sequence of ALK1 gene was identified by PCR-SSCP. The mutational segment was analyzed by DNA sequencing. An IV S4 + 3 a > t mutation was found, causing splicing abnormality of intron 4 and exon 3 skipping. CONCLUSION: A splicing pattern of the IV S4 + 3 a > t mutation has been reported among Chinese HHT2 patients for the first time. PMID- 17425870 TI - [Association of abnormality of liver enzymes and metabolic syndrome in patients with nonalcoholic fatty liver disease]. AB - OBJECTIVE: To investigate the changes of liver enzymes in the patients with nonalcoholic fatty liver disease (NAFLD) and to analyze the association between liver enzymes and metabolic syndrome (MS). METHODS: 318 non-diabetic subjects with NAFLD underwent anthropometry (height, weight, waist and hip circumference, and blood pressure), laboratory tests, including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyltransferase (GGT), serum lipid, fasting and 2 hour plasma glucose, and insulin after 75 g glucose load, and abdominal ultrasonography. Metabolic syndrome was diagnosed according to the National Cholesterol Education Program Adult Treatment Panel III criteria. RESULTS: 174 of the 318 NAFLD patients (54.7%) showed 1 to more abnormal liver enzymes. 123 of the 318 NAFLD patients (38.7%) were diagnosed as with MS, among which 48.8% of those with abnormal liver enzymes were diagnosed as with MS, a percentage significantly higher than that of those without normal liver enzyme (28.6%, P < 0.001). The levels of the 4 liver enzymes were all higher in the group with MS than in the group without MS (all P < 0.05). With the increase of the number of elevated MS components the serum levels of ALT, AST, and GGT were elevated accordingly. The risk of abnormality of liver enzyme levels significantly increased with the number of MS components after adjustment for age and sex [OR (95% CI): ALT 1.78 (1.37 approximately 2.32), AST 1.65 (1.19 approximately 2.29), GGT 1.81 (1.35 approximately 2.43)]. CONCLUSION: Most patients with NAFLD have abnormal liver enzymes, and the percentage of MS is higher in the abnormal liver enzyme group. The risk of abnormality of liver enzyme levels significantly increases with the number of MS components which suggested the components of MS might be the risk factor of increase of liver enzymes' levels. PMID- 17425871 TI - [Effects of intervention therapy of methylcobalamin and folic acid on plasma homocysteine concentration and homocysteine thiolactonases/paraoxonase activity in patients with type 2 diabetes mellitus]. AB - OBJECTIVE: To study the effects of methylcobalamin and folic acid treatment on plasma homocysteine (Hcy) level and homocysteine thiolactonase/paraoxonase (HTase/PON) activity in patients with type 2 diabetes mellitus. METHODS: 120 patients with type 2 diabetes mellitus were randomly divided into four equal groups: Group I, receiving no intervention therapy as control group, Group II, given folic acid orally (5 mg/d), Group III, receiving intramuscular injection of methylcobalamin (500 microg qd), and Group IV, treated with methylcobalamin (500 microg qd) in addition to folic acid (5 mg/d). Forty healthy age-matched persons were used as normal controls. Before and 12 weeks after 2-week treatment, plasma total Hcy, vitamin B(12), folic acid, and HTase/PON activity were assayed. RESULTS: After 12-week treatment the plasma folic acid and methylcobalamin, and Hcy levels decreased and serum HTase/PON activity increased significantly in the three groups receiving intervention treatment (all P < 0.05). The Hcy level decreased by 2.8% in Group I, 14.0% in Group II, 37.3% in Group III, and 21.7% in Group IV respectively (all P < 0.01). The HTase/PON activity increased by 2.7% in Group I, 8.0% in Group II, 3.4% in Group III, and 17.6% in Group IV respectively (all P < 0.05). CONCLUSION: Methylcobalamin and folic acid treatment alone can decrease the Hcy level and increase the HTase/PON activity in the patients with type 2 diabetes mellitus, and the methylcobalamin and folic acid combination therapy is much more effective. Folic acid may affect the HTase/PON activity through its antioxidant ability. PMID- 17425872 TI - [Association between clinical manifestations of infants with human cytomegalovirus infection and glycoprotein B genotype]. AB - OBJECTIVE: To investigate the association between the clinical manifestations of infants with human cytomegalovirus (HCMV) infection and glycoprotein B (gB) genotype. METHODS: Urine samples were obtained from 107 symptomatic infants with HCMV infection confirmed by fluorescence quantitative PCR, 70 male and 37 female, aged 5 d-8 months, and 25 asymptomatic infants with HCMV infection, 16 male and 9 female, aged 21 d-7 months. A fragment of glycoprotein B gene was amplified by nested PCR (nPCR). HCMV gB genotyping was carried out by restriction fragment length polymorphism (RFLP), and some of the amplified DNA fragments were verified by DNA sequencing. RESULTS: Of the HCMV specimens from 107 infants, 53 were typed as gB group I, 20 as gB II, 18 as gB III, 7 as gB I and gB II, 5 as gB I and gB III, and 4 as gB II and gB III, and gB IV was not found. The HCMV gB genotype from 53 infants with hepatic function damage showed that 36 were classified as gB I, 5 as gB II, 7 as gB III, 3 as gB I and gB II, and 2 as gB I and gB III. The gB I genotype was more common among the infants with hepatic function damage (P < 0.05). The distribution of gB genotype in the asymptomatic infants was as follow: gB I, 10/25; gB II, 6/25; gB III, 8/25; and gB I and gB II, 1/25. The homology of PCR products of HCMV gB in 24 strains amplified compared with the sequences of prototype strains in GenBank was from 97.06% to 99.64%. CONCLUSION: RFLP analysis of HCMV gB genotype is definite and reliable. The gB I genotype is more common among the infants with hepatic function damage. PMID- 17425873 TI - [Clinical features of thyrotoxic heart disease: analysis of 75 cases]. AB - OBJECTIVE: To investigate the clinical features of thyrotoxic heart disease. METHODS: The clinical data of 75 patients with thyrotoxic heart disease, 30 males and 45 females, aged 54.3 +/- 14.4 (17 approximately 84, with the course from 1 month to 30 years, were analyzed retrospectively. RESULTS: Forty-seven of the 75 patients (63%)presented heart failure, 47 patients (63%) showed atrial fibrillation, 2 patients (3%) showed sick sinus syndrome; 48 patients (66%) showed left atrial enlargement, 31 (43%) showed left ventricular enlargement, 30 (41%) showed right atrial enlargement, 35 (47%) showed right ventricular enlargement, and 18 (25%) showed entire heart enlargement. Mitral valve regurgitation occurred in 45 patients, tricuspid regurgitation in 45, aortic valve regurgitation in 34, and pulmonary artery hypertension (with the pulmonary blood pressure of 44 +/- 7 mm Hg and LVEF of 60% +/- 19%) in 33 patients. CONCLUSION: The clinical manifestations of thyrotoxic heart disease are various Thyroid function should be investigated to exclude thyrotoxic heart disease when routine treatment is ineffective in heart disease. The key point of treatment is hyperthyroidism. PMID- 17425875 TI - [Comparison of stenting with sirolimus eluting stent and spot stenting with small vessel stent in treatment of small vessels with long lesions]. AB - OBJECTIVE: To evaluate the short and long-term outcomes between stenting with sirolimus eluting stent (SES) and spot stenting with small vessel stent (SVS) for treatment of small vessels with long lesions. METHODS: 306 coronary heart disease patients in need of stenting in small vessel (diameter < 3.0 mm) with long lesions (> 20 mm) were divided into 2 groups: SES group (n = 93, receiving 157 CYPHER stents) and SVS group (n = 113, receiving spot stenting tactics). The clinical characteristics, success rate of procedure, rate of in-stent restenosis, target lesion revasculation (TLR), and major adverse cardiac events (MACE) were recorded after 6-month follow-up. RESULTS: The baseline clinical and angiographic characteristics were similar between these two groups. Two CYPHER stents failed to pass the tortuous and calcified lesions in the SES group; DRIVER stents were used instead and succeeded in the passing. The rates of in-stent restenosis, TLR, and MACE of the SES group were 4.0%, 2.2%, and 3.2% respectively, all significantly, lower than those of the SVS group (26.5%, 10.6%, and 13.3% respectively, all P < 0.05). CONCLUSION: In treatment of small vessel with long lesions, the rates of in-stent restenosis, TLR, and MACE are all much lower in the SES group than in the SVS group. Spot stenting with SVS may be feasible for small vessels with long and tortuous lesions, especially for those in which SES fails to pass through. PMID- 17425876 TI - [New method of lymph node tracing in gastrectomy for advanced gastric carcinoma]. AB - OBJECTIVE: To develop a new method of lymph node tracing in gastrectomy for advanced gastric carcinoma (AGC). METHODS: Forty-six patients with gastric angle carcinoma with metastasis in the third group lymph nodes underwent radical operation. During the operation methylene blue was injected inside or around the border of the metastatic third group lymph nodes. The secondary sentinel lymph nodes (SSLNs) that were the nearest to the third group lymph nodes were searched in 5 approximately 10 minutes, and the SSLNs discovered were resected to undergo HE staining and immunohistochemistry. RESULTS: SSLNs were found in 38 patients with a positive rate of 82.6% by methylene blue staining. Pathological examination found SSLNs in 30 cases with a positive rate of 78.9%. Totally 68 SSLNs were found in the 38 patients. The numbers of methylene blue positive SSLN in the groups 7, 5, 4, 6, 9, and 8 were 40, 9, 7, 5, 4, and 3 respectively. Pathology and histochemistry found 31, 9, 5, 4, 2, and 1 SSLNs in the groups 7, 5, 4, 6, 9, and 8 with the positive rates of 77.5%, 100%, 71.4%, 80.0%, 50.0%, and 33.3% respectively. There were no significant correlation between the size and degree of differentiation of tumor and the positive rate of SSLN. CONCLUSION: The technique of SSLN tracing expands the application range of SLN tracing and provides a new thinking for study of lymph node in gastric carcinoma in different stages. PMID- 17425878 TI - [Anti-apoptotic effects of mesenchymal stem cells on cardiac myocytes: in vitro study with rats]. AB - OBJECTIVE: To investigate the anti-apoptotic effects of mesenchymal stem cells (MSCs) on hypoxia-injured cardiac myocytes. METHODS: MSCs were isolated from the bone marrow of 6 - 8 week-old Sprague-Dawley rats. Cardiac myocytes from neonatal rat were cultured under hypoxia, then the hypoxia-injured cardiac myocytes were divided into 3 groups: cultured alone (control group), co-cultured with the MSCs, or co-cultured in MSC-conditioned media in conditions of anoxia (95% N(2) + 5% CO(2), continuous hypoxia group) or normoxia [hypoxia/reoxygen (H/R) group] for 72 hours. The cell apoptosis was measured by Hoechst staining, and Western blotting was used to test the protein expression of Bcl-2 and Bax in the cardiac myocytes. RESULTS: The apoptotic rate of the cardiac myocytes cultured under hypoxia was 51.6% +/- 2.4%, significantly higher than those of the cardiac myocytes co-cultured with MSCs and MSC-conditioned media respectively (15.1% +/- 5.4% and 24.0% +/- 4.2% respectively, both P < 0.001). The apoptotic rate of the H/R group was 20.9% +/- 2.7%, significant higher than that of the MSC group (11.5% +/- 3.7%, P < 0.05), however, not significantly different from that of the MSC-conditioned media group (20.1% +/- 4.2%, P > 0.05). The protein expression of Bcl-2 was not significantly different among different groups. The Bax protein expression of the MSC group and MSC-conditioned media group were 2.28 +/- 0.46 and 3.01 +/- 0.26 respectively, both significantly lower than that of the control group (7.62 +/- 1.28, both P < 0.05). The decreased expression of Bax in the cardiac myocytes was greatly related to the decreasing of apoptosis. CONCLUSION: Co-cultured MSCs show significant anti-apoptotic effects on cardiac myocytes both in continuous hypoxia and in H/R conditions with the possible mechanism of direct cell to cell interaction and paracrine of cytokines which effect the expression of Bax in the myocytes. PMID- 17425879 TI - [Long-term effects of recurrent seizures in neonate period on gamma-aminobutyric acid A receptor alpha1 and beta2 subunits expression in adult brain: experiment with rats]. AB - OBJECTIVE: To investigate the long-term effects of recurrent seizures in neonate period on the expression of gamma-aminobutyric acid A receptor (GABAAR) alpha1 and beta2 subunits in brain and spatial memory and seizure susceptibility in adult period. METHODS: Thirty-two 7-day-old SD rats were randomly divided into 2 equal groups: seizure group, inhaling flurothyl to induce seizure daily for 6 days, and control group. On days 61 - 65 after birth Morris water maze test was used to record the escape latency. On day 75 after birth pentylenetetrazol (PTZ) was injected intraperitoneally to induce seizure so as to record the latency. Then the rats were killed to take their brains, 8 in each group used to undergo immunohistochemistry to examine the protein expression of the GABAAR alpha1 and beta2 subunits, and the other 8 in each group used to examine the mRNA expression of the GABAAR alpha1 and beta2 subunits in the brains using RT-PCR. RESULTS: On day 64 the escape latency of the seizure group was 82,424 ms +/- 35,622 ms, significantly longer than that of the control group (40,712 ms +/- 29,468 ms, P = 0.001). On day 75 the frequency of crossing target within 120 s in the water maze of the seizure group was 1.2 times +/- 0.9 times, significantly less than that of the control group (3.1 times +/- 1.3 times, P < 0.01). The seizure latency after the PTZ injection of the seizure group was (1487 +/- 662) s, not significantly different from that of the control group (1841 s +/- 648 s, P = 0.133). In comparison with the control group the accumulated optical density (AOD) of GABAAR alpha1 subunit protein immunoactivity in the parietal cortex, and hippocampal CA1 2 and CA4 regions of the seizure group decreased significantly (all P < 0.05), and was not significantly different in the frontal cortex, dentate gyrus, and hippocampal CA3 region (all P > 0.05). In comparison with the control group the accumulated optical density (AOD) of GABAAR beta2 subunit protein immunoactivity in the thalamus, and hippocampal CA1-4 regions of the seizure group decreased significantly (all P < 0.05), and was not significantly different in the frontal cortex and parietal cortex (both P > 0.05). In comparison with the control group the mRNA expression of GABAAR alpha1 subunit and the mRNA expression of GABAAR beta2 subunit of the seizure group were significantly lower in the hippocampus (both P < 0.05) and not significantly different in the cerebral cortex (both P > 0.05). CONCLUSION: Recurrent seizures in neonate period modify the expression of GABAAR alpha1 and beta2 subunits in the cerebral cortex and hippocampus in adult period which may be related to cognitive deficit. PMID- 17425885 TI - [Scientific productivity and Hirsch's h index of Spanish social psychology: convergence between productivity indexes and comparison with other areas]. AB - This article describes Hirsch's h index as a measure of researchers' scientific productivity and quality, examining its convergent validity with other productivity and quality measures (total citations, maximum citation, sexennials, years since earning Ph.D., and years as a full professor). It shows the distribution of the index among Spanish Social Psychology professors, and a comparison is made among the ten professors with highest h index of the six knowledge areas into which academic Psychology is divided in Spain. The results show that the index largely correlates with the citations but only very modestly with the number of sexennial investigations. The distributions by areas show that Psychobiology and Basic Psychology, as a whole, have a higher h index than the other areas, with Social Psychology in fourth place. Lastly, we discuss the implications of these results and we offer some recommendations for using the index in academic and research decision making. PMID- 17425886 TI - [Evaluation of communicative and linguistic development using the Spanish version of the MacArthur-Bates inventories]. AB - The MacArthur-Bates Inventories are a valid and reliable method for assessing communicative and linguistic skills in infants (8-15 months) and young children (16-30 months), and have been adapted to many languages. This paper presents their adaptation to Spanish. Structure, innovations included in the Spanish version, and the standardisation process are described, and some results obtained from the analysis of a sample of 1150 children are presented. The advantages and limitations of the use of parental report inventories are highlighted. For children before three years of age, there are very few assessment tools of language and communicative development with sufficient guarantees produced and edited in Spain. The Spanish version of the MacArthur-Bates Inventories represents an important contribution for early identification of language delays or disorders, and for follow-up of these cases. PMID- 17425887 TI - [Influence of the recording interval and a graphic organizer on the writing process/product and on other psychological variables]. AB - An experimental study of the influence of the recording interval and a graphic organizer on the processes of writing composition and on the final product is presented. We studied 326 participants, age 10 to 16 years old, by means of a nested design. Two groups were compared: one group was aided in the writing process with a graphic organizer and the other was not. Each group was subdivided into two further groups: one with a mean recording interval of 45 seconds and the other with approximately 90 seconds recording interval in a writing log. The results showed that the group aided by a graphic organizer obtained better results both in processes and writing product, and that the groups assessed with an average interval of 45 seconds obtained worse results. Implications for educational practice are discussed, and limitations and future perspectives are commented on. PMID- 17425888 TI - [Individual differences in hypothetic-deductive reasoning: importance of cognitive skills and flexibility]. AB - Recent investigations show that people's differential performance when solving reasoning tasks is due to differences not only at computational level, but also at the rational or intentional level. This study is within the framework of this context and it specifically analyses individual differences in hypothetic deductive reasoning, taking into account the characteristics associated with the task itself (content and instructions) as well as the individuals' differential characteristics. Two hundred and seventy-six students from the University of Santiago de Compostela participated in this study. Several psychometric tests were administered and participants were requested to solve various reasoning tasks. The results obtained confirm that, in effect, the differences between competent and non-competent reasoners are observed at both the above-mentioned levels: cognitive skills and abilities (algorithmic level) and cognitive flexibility (intentional level). PMID- 17425889 TI - [Complexity and social psychology of organizations]. AB - This article presents complexity (nonlinearity, chaos, self-organisation, fractals...) as a new and emerging epistemological paradigm, an alternative to an old, simpler, and reductionist paradigm. According to this point of view, we try to view work organizations as complex adaptive systems (CAS). Lastly, we offer a review of the literature on the applications of complexity to Organizational Social Psychology. PMID- 17425890 TI - [Psychosocial risk factors at work as predictors of mobbing]. AB - This work analyses the way in which various psychosocial risk indicators may predict mobbing. A sample of 638 workers, 168 men and 470 women, from the fruit and-vegetable sector was evaluated. An anonymous questionnaire was administered to all employees who were present on the evaluation days in the companies comprising the study. After analysing the data obtained with the mobbing questionnaire NAQ-RE (Saez, Garcia-Izquierdo, and Llor, 2003) and with the psychosocial risk factors evaluation method of the INSHT (Martin and Perez, 1997), using canonical regression, we found that several psychosocial factors such as role definition, mental workload, interest in the workers, and supervision / participation predict two types of mobbing: personal mobbing and work-performance-related mobbing. PMID- 17425891 TI - [Culture and cultural gaps in work teams: implications for organisational commitment]. AB - Some theoreticians of organisational commitment have proposed that culture is an important determinant of organisational commitment. Nevertheless, very few studies have examined the role that work teams culture (subculture) and their cultural gaps play in commitment. This study is an attempt to overcome this lack. Using a sample of 375 work teams from various public and private organisations, it was found that the results confirmed our proposals. Cultural gaps were negatively related to commitment; the teams subculture was positively related to commitment, and more highly to commitment to values than to commitment to continuing. Contrary to the results of other studies, the demographic variables (age, time on the team, time in the company) were not significant, except that educational level was related to the commitment to continue. The implications of these results are analysed. PMID- 17425892 TI - Supervisors safety response: a multisample confirmatory factor analysis. AB - Supervisors Safety Response (SSR) has been closely linked to workplace safety, and, as perceived by workers, it is considered to be one of the most influential issues with regard to employees compliance with safety behaviours. This study defines and tests a bifactorial and a monofactorial model of the SSR. Two facets of the SSR were measured: (a) supervisors response toward workers safe or unsafe behaviour and (b) supervisors safety attitudes and behaviours applied to their own work. In three samples of injured blue-collar workers ( N(1) = 110, N(2) = 123, N(3) = 104), multisample confirmatory factor analyses, using maximum likelihood estimation, were conducted to test both the bifactorial and the monofactorial model. Both models provide an overall good fit, but parsimony and the high correlation between factors in the bifactorial model support the monofactorial model. Attention is drawn to the measurement of the SSR as a diagnostic tool useful in selecting intervention goals, specifically integrating supervisors safety behaviour. PMID- 17425893 TI - Picture priming in normal aging and Alzheimer's disease. AB - The present study investigated age invariance for naming pictures and whether implicit memory is spared in Alzheimer's disease (AD). During the study phase, young adults, AD patients, and older controls were shown outlines of familiar pictures. After a distracter task, implicit memory was assessed incidentally. The results showed similar visual priming for the three groups, although young adults responded faster than the two older groups. Moreover, the number of errors was smaller for studied than for non-studied pictures. This pattern of results was repeated across the three groups, although AD patients produced more errors than young adults and older controls, and there were no differences between these latter groups. These results confirmed previous visual and haptic findings showing unimpaired perceptual priming in normal aging and AD patients when implicit memory is assessed using identification tasks. These results are interpreted from a cognitive neuroscience perspective. PMID- 17425894 TI - [Gender and emotional response induced by imagery]. AB - The aim of this study is to explore gender differences in emotional expression: Do men benefit from their stereotyped response pattern to some negative affects such as sadness? Do women benefit less than men from positive affect? We studied sadness and happiness in the laboratory, using imagery induction with some temporal proximity, and registering physiological, facial, and cognitive responses. The results show a complex panorama in which the differences depend on the emotional content and presentation order. The results are in accordance with the educational theories that postulate prototypical emotional education, and indicate a way to reduce the problems related to women's sensitization to sadness, using the beneficial effects of positive experiences. PMID- 17425895 TI - Exercise motivation in university community members: a behavioural intervention. AB - The purpose of this study was to examine how exercise level, exercise motives, and barriers changed from the baseline phase to the follow-up phase after a behavioural and cognitive intervention aimed at increasing exercise. Seventy-five members of our university community (43 subjects in the control group and 32 in the experimental group), all of whom received cognitive feedback, agreed to complete the baseline phase. Only the experimental group received behavioural feedback and a free-access gym ticket with personal training in order to facilitate their adherence to exercise. The results suggest that a combination of behavioural and psychological techniques is an efficient strategy for increasing exercise level. In addition, the results showed that extrinsic motivation predominates the early stages of change-of-exercise behaviour, and that intrinsic motivation is important for progression towards maintenance. Subjects who decreased their exercise level increased their extrinsic exercise motivation and subjects who increased their exercise level decreased the barriers related to intrinsic motivation. These findings indicate that, in order to facilitate exercise adherence, feedback about motives for undertaking exercise is needed, combined with advice about how to improve physical condition. This combination could help eliminate certain barriers that hinder engaging in an active and healthy life-style. PMID- 17425896 TI - The commitment of young soccer players. AB - The main purpose of this study was to test the Sport Commitment Model in young soccer players. Participants' ( N = 437) age ranged from 14 to 16 years. Based on the sport commitment model proposed by Scanlan and colleagues, 6 scales and 28 items constitute the Sport Commitment Questionnaire (SCQ). Results of this study provide some support for the reliability and validity of the SCQ, using a sample of young soccer players. Reliability analysis provided acceptable internal consistency for four scales: Sport Commitment, Sport Enjoyment, Involvement Alternatives and Social Constraints. Sport Enjoyment (Beta= .56, p <.001) and Involvement Alternatives (Beta= -.40, p <.001) were the strongest sport commitment predictors. Further studies with a broader age range, different genders, and other sports or competitive levels are required to continue analyzing the sport commitment model. PMID- 17425897 TI - [Junior version of the Spanish NEO PI-R (JS NEO)]. AB - This article presents the preliminary data of the development of a Junior version of the Spanish NEO PI-R (JS NEO), suitable for teenagers from 12 to 17 years of age. From the 240 original English items, 92 were modified or reworded to some degree to make the vocabulary adequate for this age group. The psychometric properties of the JS NEO were investigated in a sample of 2,505 adolescents. Results showed that the adult NEO PI-R factor structure was replicated in the junior version of the inventory. Internal consistency and temporal stability reliabilities of the scales were adequate for most scales. Furthermore, the cross form correlations between the junior (JS NEO) and the adult (NEO PI-R) scales indicated satisfactory construct validity of the junior version of the inventory. PMID- 17425898 TI - [Personality, gender and violent criminality in prison inmates]. AB - The Antisocial Personality Questionnaire developed by Blackburn and Fawcett (1999) has shown adequate reliability and validity in studies of male offenders interned in psychiatric hospitals. This paper provides data on the APQ collected from a sample of offenders (males and females) without any diagnosed mental illness. The sample was made up of 216 offenders (108 males and 108 females) confined in Spanish prisons. We analyzed the psychometric properties of the instrument and we also examined differences in personality as a function of gender and type of crime (violent vs. non-violent). Results support the reliability of the APQ scales as well as the structure proposed by Blackburn. Additionally, we selected items that discriminate between offenders convicted for violent and non-violent crimes; when these items are factorized, a three-factor structure emerges, resembling Eysenck's model. PMID- 17425899 TI - [The consequences of a nurse's negative evaluation of a blood pressure reading. Implications for the etiology of white-coat hypertension]. AB - The aim of this study was to show how a nurses negative evaluation of a blood pressure (BP) reading can cause an increase in BP in the following reading. Twenty-five subjects with normal pressure who had been informed that they were to participate in an epidemiological BP study took part and they were divided into two groups. After taking the first BP reading, participants from the experimental group were informed that their pressure was normal but high for their age, whereas those in the control group were told that their pressure was normal for their age. The results showed that this comment caused a significant increase in the diastolic BP of the experimental group. The theoretical and clinical implications of this reaction are discussed in relation to the phenomenon known as White-Coat Hypertension from the perspective of the theory of incubation. PMID- 17425900 TI - Implementation intentions: control of fear despite cognitive load. AB - The cognitive consequences of forming implementation intentions in controlling fear were addressed in the present study. Participants with an intense fear of spiders evaluated pictures of spiders, pleasant pictures, and neutral pictures under cognitive load. Regulatory control was measured by participants' self report ratings of the pictures on the Self-Assessment Manikins Scales. Only participants given implementation intentions reported weaker negative emotional responses to the pictures of spiders as compared to participants given a goal intention and to no-goal control participants. Thus, emotional control by implementation intentions was shown not to tax a person's cognitive resources, attesting to the automatic nature of this self-regulation strategy. PMID- 17425901 TI - [Depression and the complete state model of health]. AB - Health is a state of complete physical, mental, and social well-being and not merely the absence of disease or infirmity. In order to specify the contents of this positive state, the Complete State Model of Health (CSMH) considers mental health as a series of symptoms of hedonia and positive functioning, operationalized by measures of subjective, psychological, and social well-being. This model has empirically confirmed two new axioms: (a) rather than forming a single bipolar dimension, health and illness are correlated unipolar dimensions, and (b) the presence of mental health implies positive personal and social functioning. In the present article, we have taken the CSMH as the theoretical framework for the study of depression. Confirmatory factor analyses did not support the first axiom. In fact, the model that posits that measures of mental illness and health form a single bipolar dimension provided the best fit to the data. PMID- 17425902 TI - Brain c-Fos immunocytochemistry and cytochrome oxidase histochemistry after a fear conditioning task. AB - The involvement of the basolateral and the medial amygdala in fear conditioning was evaluated using different markers of neuronal activation. The method described here is a combination of cytochrome oxidase (CO) histochemistry and c Fos immunocytochemistry on fresh frozen brain sections. Freezing behavior was used as an index of auditory and contextual fear conditioning. As expected, freezing scores were significantly higher in rats exposed to tone-shock pairings in a distinctive environment (conditioned; COND), as compared to rats that did not receive any shocks (UNCD). CO labeling was increased in the basolateral and medial amygdala of the COND group. Conversely, c-Fos expression in the basolateral and medial amygdala was lower in the COND group as compared to the UNCD group. Furthermore, c-Fos expression was particularly high in the medial amygdala of the UNCD group. The data provided by both techniques indicate that these amygdalar nuclei could play different roles on auditory and contextual fear conditioning. PMID- 17425903 TI - [Language is not neutral. Commentary about APA style]. AB - Some basic characteristics, not always explicit, of the editorial style proposed by the American Psychological Association (APA), and from the objections posed by some authors who maintain critical positions towards the use of this style, are reviewed, starting with the work of Madigan, Johnson, and Linton (1995) and the subsequent controversy. Starting with this review, problems related to underlying assumptions of the style, with ethical aspects of research, and with the epistemological positions defended by the different traditions of research are discussed. In the conclusions, a simpler differentiation between the scientific technical and communicative-practical systems of enquiry is proposed, and an explicit commitment, in the text of the report, to the ethical responsibilities derived from the authorship and the development of the research. PMID- 17425904 TI - [How to fit and interpret multilevel models using SPSS]. AB - Hierarchic or multilevel models are used to analyse data when cases belong to known groups and sample units are selected both from the individual level and from the group level. In this work, the multilevel models most commonly discussed in the statistic literature are described, explaining how to fit these models using the SPSS program (any version as of the 11 th ) and how to interpret the outcomes of the analysis. Five particular models are described, fitted, and interpreted: (1) one-way analysis of variance with random effects, (2) regression analysis with means-as-outcomes, (3) one-way analysis of covariance with random effects, (4) regression analysis with random coefficients, and (5) regression analysis with means- and slopes-as-outcomes. All models are explained, trying to make them understandable to researchers in health and behaviour sciences. PMID- 17425905 TI - [Application of the pi* goodness-of-fit index to latent structure models]. AB - Testing model fit for latent structure models (latent trait models and latent class models) is difficult because of the lack of goodness-of-fit statistics with a known distribution. This paper describes the application of the pi* goodness-of fit statistic to latent structure models. The statistic pi* is based on the concept of latent classes and has a natural interpretation when applied to these models. This statistic assumes that the population is made up of several classes that follow a parametric model, and a residual class outside the model. The value of pi* is the population proportion in the residual class. This paper describes the estimation algorithms of pi* for latent trait and latent class models and an empirical example with a scale of study habits. There are two latent classes in these data: bad and regular students, which are related to the student s responsibility. PMID- 17425906 TI - [Application of four procedures for detecting differential item functioning in polytomous items]. AB - The authors describe and use four methods for detecting Differential Item Functioning in polytomous items: Mantel, Generalized Mantel-Haenszel (GMH), Ordinal Logistic Regression (RLO), and Discriminant Logistic Regression (RLD). For each procedure, the theoretical model and the measure of effect size are described. The data from the "Reading Comprehension Test" from the PISA2000 evaluation program were analyzed using a cross-validation design. Two booklets were independently evaluated in the American and Spanish samples. Adopting as decision rule the significance of the statistical test and the measurement of the effect size, agreement among the evaluated procedures was total for two of the analyzed items. PMID- 17425907 TI - [Item characteristics that predict response latency in personality measures]. AB - This study assesses some item characteristics that are expected to affect response latencies in the computerized administration of a personality questionnaire. First, some previous empirical results are discussed, and a theoretical framework from which predictions can be made is proposed. Second, the predictions are empirically assessed using two data-sets, one of them based on binary items, and the other one based on graded-response items. Some of the results obtained are new, whereas the remaining agree with previous empirical results. The implications of the present results in applied research, and particularly for the use of latencies, is discussed. PMID- 17425908 TI - [Interaction in ANOVA: misconceptions]. AB - Although interaction in analysis of variance has an unequivocal theoretical meaning (and so it appears in the statistic literature), frequent misconceptions are found in empirical research, which, in many cases, lead to wrong conclusions. In this paper, 150 articles are reviewed: in 12.7% of them, no attention is paid to the interaction (either because it is not analysed or because it is analysed but not discussed); in 79.1%, interaction is studied through simple effects analysis; and only in 8.2% of the cases, interaction is correctly discussed. It could be that psychology researchers tend to analyse and interpret the interaction between factors incorrectly because the most widespread statistic packages (with SPSS in the lead) do not allow performing the comparisons needed to analyse a significant interaction in factorial designs with randomized groups. In order to contribute to eradicating this problem, we herein show how to design some of the linear comparisons that allow isolating the interaction effect, and we explain how to use SPSS to compute these comparisons. PMID- 17425909 TI - [Structural validity of miokinetic psychodiagnosis-revised and digitalyzed (MKP RD) lineograms]. AB - In this study, we have applied a principal components analysis to the data obtained from a graphical record of motor behaviour performed without vision. The data was obtained from two samples, one made up of men and the other one made up of women. Hand dominance was controlled in both samples. Five independent factors were obtained: Factor I (Tendency to increase or decrease the length), Factor II (Tendency to deviate in a sagittal direction), Factor V (Tendency to deviate in a frontal direction); two factors (III and IV) were found in a transversal direction. This factor structure did not depend on sex, because the same structure was found in both sexes, nor it could be explained by learning, because the same factors appeared for each hand, except for Factors III and IV which were different for each hand. This study supports different motor dispositions as foundations of behaviour tendencies. PMID- 17425910 TI - The role of PPAR-alpha agonists in the prevention of CVD in diabetes. PMID- 17425911 TI - On the way to a diabetes cure? Improvement of glycemic control through islet transplantation with the use of a standardized common protocol: international trial of the Edmonton protocol for islet transplantation. PMID- 17425912 TI - Mucosal exposure to antigen: cause or cure of type 1 diabetes? AB - The human gut offers more than 200 m2 of mucosal surface, where direct interactions between the immune system and foreign antigens take place to eliminate pathogens or induce immune tolerance toward food antigens or normal gut flora. Therefore, mucosally administered antigens can induce tolerance under certain circumstances. In autoimmune diabetes, mucosal vaccination with autoantigens elicits some efficacy in restoring tolerance in mice, but it never succeeded in humans. Furthermore, in some instances autoimmunity can be precipitated upon oral or intranasal autoantigen administration. Therefore, it is difficult to predict the effect of mucosal vaccination on autoimmunity and much effort should be put into establishing better assays to reduce the risk for possible adverse events in humans and enable a rapid and smooth translation. PMID- 17425913 TI - Randomized controlled clinical trials of psychological interventions to improve glycemic control in pediatric and adult patients with T1DM. PMID- 17425914 TI - Technology and intensive management in youth with type 1 diabetes: state of the art. AB - Recent implementation of technologic advances in the management of pediatric type 1 diabetes creates an exciting time for the diabetes community. Safety and efficacy of new therapies are commonly demonstrated first in adult patients with pediatric studies following. This review covers recent advances in six main areas: blood glucose monitoring including continuous monitoring; point-of-care A1c testing; designer insulins; insulin delivery systems including insulin pumps; electronic communication; and telehealth. PMID- 17425915 TI - Recent advances in understanding depression in adults with diabetes. AB - The authors review the science linking depression with diabetes. Some recent heuristic research is identified that highlights progress in the field and is directing future research. Issues in the management of depression in diabetes are outlined, including interactions of depression and insulin resistance. PMID- 17425916 TI - Efficacy of lifestyle behavior change programs in diabetes. AB - Evidence that a comprehensive lifestyle behavior change program targeting eating and physical activity, and ultimately weight loss, can alter the course of diabetes progression is growing. This report summarizes data from major clinical trials primarily, describes key features of behavior change programs, and comments on existing challenges regarding the adequacy, sustainability, and feasibility of behavior change programs for weight loss and diabetes prevention. PMID- 17425917 TI - Insights on pathogenesis of type 2 diabetes from MODY genetics. AB - Maturity-onset diabetes of the young (MODY) is a type of non-insulin-dependent diabetes mellitus caused by rare autosomal-dominant mutations. MODY genes play key biochemical roles in the pancreatic beta cell; therefore, common variants of MODY genes are excellent candidate genes for type 2 diabetes. We review recent studies that suggest that common MODY gene variation contributes modestly to the heritability of type 2 diabetes. PMID- 17425918 TI - Gene discovery in diabetic nephropathy. AB - Diabetic nephropathy affects approximately one third of diabetic patients and is the leading cause of end-stage renal disease. Identification of genetic susceptibility factors contributing to the development of diabetic nephropathy could facilitate prediction, development of improved treatments, and prevention of this devastating complication of diabetes. Although investigations to identify the causal genetic variants associated with diabetic nephropathy have been inconclusive, new approaches, including whole genome association scanning, offer hope for the future. PMID- 17425919 TI - The biology of RAGE and its ligands: uncovering mechanisms at the heart of diabetes and its complications. AB - The interaction of glucose-modified and inflammation-promoting ligands with the receptor for advanced glycation end products (RAGE) is emerging as a central mechanism contributing to the diverse complications of diabetes. These ligands, particularly in oligomeric form, bind to RAGE and transduce intracellular signals. The consequences of this interaction, as elucidated in cultured cells and animal models, include upregulation of inflammatory and tissue-degradative pathways. Pharmacologic antagonism of RAGE may hold promise for the treatment of diabetic complications. PMID- 17425920 TI - The search for type 2 diabetes susceptibility loci: the chromosome 1q story. AB - The unbiased approach of genome-wide linkage analysis has shown evidence for linkage of type 2 diabetes mellitus to the chromosome 1q21-25 region in at least eight independent studies. More than 26 candidate genes have already been evaluated, but to date none explain the evidence for linkage in this gene-dense region. Considerable data suggest that multiple genes account for this linkage result. The search for these genes is now the focus of an international consortium of groups reporting linkage of type 2 diabetes to this region of chromosome 1q21-q25. PMID- 17425921 TI - [No resuscitation in heart arrest--a hidden decision?]. PMID- 17425922 TI - [Do-not-resuscitate orders--ethical and legal considerations]. AB - The article is based on a structured literature review. Practicing DNR orders in Denmark has not yet been described and there are no national guidelines. Generally patients must give informed consent before do-not-resuscitate (DNR) orders. Only 32% of competent patients are involved prior to the decision even though patients wish to talk about decisions regarding the end of life . In general doctors cannot predict patients' do-not-resuscitate preferences and doctors underestimate patients' self-estimated quality of life. Doctors rarely contact the Danish Living Will Registry. Patients with do-not-resuscitate orders may be subject to other therapeutic reductions. In conclusion, patient autonomy should also be respected in the case of do-not-resuscitate orders. PMID- 17425923 TI - ["Do not resuscitate" orders in Danish medical wards. A questionnaire]. AB - INTRODUCTION: Do not resuscitate (DNR) orders should prevent pointless life prolonging procedures. Practicing DNR orders in Denmark is not described and there are no national guidelines. The purpose of this study is to clarify how DNR orders, patient autonomy, and living wills are handled in a Danish medical ward. MATERIALS AND METHODS: All 193 medical wards in Denmark received questionnaires (Figure 1) addressed to the medical Head of Department. 138 (71.5 %) questionnaires were analysed. RESULTS: 127 (92 %) wards had DNR orders. In 52 (38 %) wards DNR orders could include reduction of other treatment modalities. Competent patients were 'always' asked in 20 (14 %), 'often' in 34 (25 %), 'seldom' in 59 (43 %), and 'never' in 12 (9 %) of the wards prior to DNR decisions. Spouses were asked more often than the patient; 'always' in 31 (22 %), 'often' in 68 (49 %), 'seldom' in 21 (15 %), and 'never' in 4 (3 %) of the wards. The Danish Living Will Registry was contacted 'always' in 2 (1 %), 'often' in 15 (11 %), 'seldom' in 68 (49 %), and 'never' in 37 (27 %) of the wards. 112 (81 %) wards did not have a written guideline on DNR orders. CONCLUSION: Patients should be asked more often prior to DNR decisions. Health workers should contact The Danish Living Will Registry more frequently. As a decision aid and in order to strengthen both health workers' and patients' legal rights, a national guideline on DNR ordering could be established. PMID- 17425924 TI - [Stress induced "Takotsubo" cardiomyopathy: you can die from a broken heart!]. AB - Stress induced "Takotsubo" cardiomyopathy is a recently described cardiovascular syndrome mimicking acute myocardial infarction and characterised by reversible left ventricular dysfunction with apical ballooning in patients without coronary artery disease. It usually affects elderly women and is preceded by emotional or physical stress. With early diagnosis and aggressive therapy, the prognosis is favourable. The syndrome is probably underdiagnosed, and the pathophysiology may involve coronary vascular dysfunction, excessive sympathetic stimulation and left ventricular outflow obstruction. PMID- 17425925 TI - [Surgical versus non-surgical treatment for femoral pseudoaneurysms]. PMID- 17425926 TI - [Excision of skin tumor]. PMID- 17425927 TI - [Uniform reference intervals and harmonised results in clinical biochemistry in Scandinavia--light ahead]. AB - Danish laboratories are introducing identical reference intervals for a number of biochemical components in accordance with a Nordic agreement and recommendations during 2007. Danish doctors will consequently experience both changes in reference interval limits as well as adjustments of patients' levels of reported results for some components. PMID- 17425928 TI - [Scientific qualifications of younger physicians in residency in Denmark 1998 2003]. AB - INTRODUCTION: The purpose of this study is to describe the scientific qualifications of younger physicians in residency in Denmark during 1998-2003. MATERIALS AND METHODS: Data consist of all applications for residency during 1998 2003 in 21 medical specialties. 1920 physicians in residency are included in the study. Connections between scientific activities and the residents' specialty and education (university, foundation year (1 year), and first year of specialist training) are investigated. RESULTS: 74% of young physicians in residency have scientific qualifications and 16% have a PhD or a doctor's degree. Scientific qualifications vary much between specialties. Physicians graduated from Aarhus University have higher scientific qualifications than physicians educated from the other universities; in essence because more have a PhD. Physicians with first year of specialist training from a university hospital have higher scientific qualifications than physicians with training from other hospitals. Foreign physicians have low scientific production. Physicians with a PhD or a doctor's degree have a significantly higher candidate age than the average for all residents, but it does not equal the time a PhD takes. CONCLUSIONS: No reduction in residents' total scientific qualifications were found in the period 1998-2003, but there is a tendency towards greater spread in scientific qualifications among the residents. Specialty, university and first year of specialist training are all factors that influence young physicians' scientific activities. PMID- 17425929 TI - [Gender, qualifications and choice of specialty of younger physicians in Denmark 1998-2003]. AB - INTRODUCTION: The purpose of this study is to describe differences in specialty choices and qualifications between male and female physicians at the start of their residency in Denmark 1998-2003. MATERIAL AND METHOD: Data consist of all applications for residency in the period 1998-2003 in 21 medical specialties. 1,920 physicians in residency are included in the study. Their gender is compared with specialty choices, age and points achieved on supplementary criteria. RESULTS: Over 80% of the residents in child and youth psychiatry and gynaecology/obstetrics were women. 87% of the residents in orthopaedic surgery were men. No gender differences were found in biological age and candidate age at the start of residency. Women achieved higher points for clinical qualification and theoretical courses, while the male residents achieved the highest points for scientific and pedagogical qualification. Within the individual medical specialties there were no gender differences. CONCLUSION: Gender does not have independent influence on achieved residency, but the study shows that there are gender differences in specialty choice and in the way men and women qualify for residency. The clear differences between men and woman across medical specialties are not matched by similar differences within each specialty. Gender differences thus seem determined by differences in specialty choice. The study results can help create the basis for the future medical residency and can guide the considerations of specialty societies, young physicians and other interested parties with regard to recruitment and specialty profile. PMID- 17425930 TI - [Neonatal hypernatraemic dehydration--why increasing incidence?]. AB - INTRODUCTION: Neonatal dehydration with hypernatraemia is a serious condition with risk of cerebral damage and death. Recent studies have reported a rising incidence. MATERIALS AND METHODS: A retrospective study was conducted at Hvidovre Hospital over a 5-year period to identify term or near-term infants (>35 weeks of gestation) who were admitted with a weight-loss >10%. Infants admitted between the 3rd and the 14th day of life with a discharge diagnosis code indicating dehydration were also included. RESULTS: During the period a total of 89 infants were admitted and 24 had hypernatraemia. The incidence increased from 2.1 to 4.9 (RR 2,5-p >0.0008). All infants were breastfed and only 3 had had supplementation prior to admission. The weight-loss ranged between 270 and 1100 grams (equivalent to 10.1-29.7% reduction in birth weight) while lethargy and jaundice were frequent symptoms. Cerebral complications occurred for 8 infants and 2 developed permanent brain damage. CONCLUSIONS: Breastfeeding should be encouraged but supplementation may be necessary for some infants. Infants born to primiparous women, infants heavy for gestational age and infants who do not thrive on day 4-5 appear to have a high risk. These infants could be identified by regular weighing and, if necessary, supplementation and follow-up should be instituted. PMID- 17425931 TI - [Severe hypernatraemic dehydration in collodion baby]. AB - Case report on severe hypernatraemic dehydration in a non-recognised collodion baby who also suffered from hydrops fetalis caused by supraventricular tachycardia. Excessive transcutaneous fluid loss caused s-Na+ reaching 182 mmol/l within 36 hours of birth. The infant was cautiously rehydrated during the following three days. No sign of neurologic impairment was observed. It is emphasized that early observation of the collodion baby must take place in a humidified incubator. Major weight changes in the newborn should always result in analysis of serum sodium. PMID- 17425932 TI - [Losartan-induced Stevens-Johnson syndrome in a geriatric patient]. AB - Stevens-Johnson syndrome is an acute inflammatory and reactive disorder with skin and severe mucous membrane changes. Often patients have associated constitutional symptoms, including fever. The syndrome can be precipitated by infection or medication. We report a case of losartan-induced Stevens-Johnson syndrome associated with renal impairment in a geriatric patient. PMID- 17425933 TI - Risk factors associated with the prevalence of tuberculosis-like lesions in fenced wild boar and red deer in south central Spain. AB - In recent decades the management of large game mammals has become increasingly intensive in south central Spain (SCS), resulting in complex epidemiological scenarios for disease maintenance, and has probably impeded schemes to eradicate tuberculosis (TB) in domestic livestock. We conducted an analysis of risk factors which investigated associations between the pattern of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red deer (Cervus elaphus) across 19 hunting estates from SCS and an extensive set of variables related to game management, land use and habitat structure. The aggregation of wild boar at artificial watering sites was significantly associated with an increasing risk of detecting TBL in both species, which probably relates to enhanced opportunities for transmission. Aggregation of wild boar at feeding sites was also associated with increased risks of TBL in red deer. Hardwood Quercus spp. forest availability was marginally associated with an increased risk of TB in both species, whereas scrubland cover was associated with a reduced individual risk of TBL in the wild boar. It is concluded that management practices that encourage the aggregation of hosts, and some characteristics of Mediterranean habitats could increase the frequency and probability of both direct and indirect transmission of TB. These findings are of concern for both veterinary and public health authorities, and reveal tuberculosis itself as a potential limiting factor for the development and sustainability of such intensive game management systems in Spanish Mediterranean habitats. PMID- 17425934 TI - A descriptive spatial analysis of bovine tuberculosis in intensively controlled cattle farms in New Zealand. AB - We describe the temporal and geographical distribution of confirmed cases of bovine tuberculosis (TB) in a population of cattle in the south-east of the North Island of New Zealand. Data were derived from routine TB testing conducted between 1980 and 2003 and included details for 69 farms. Four six-year periods were defined to coincide with changes in depopulation strategies against the wildlife TB reservoir, the brushtail possum Trichosurus vulpecula. For the periods 1980 to 1985 and 1986 to 1991 the median annual incidence rate of TB was 0.4 and 4.7 cases per 1000 cattle-years at risk, respectively. For the period 1992 to 2003 the median annual incidence rate of TB decreased to 1.8 cases per 1000 cattle-years at risk, coincident with the use of poisoning to control possums in the surrounding forest park (a major possum habitat area). We identified clusters of TB cases adjacent to the forest park and found no evidence of spatio-temporal interaction of TB risk among farms. Our findings support the hypothesis that possums living in the forest park are a source of bovine TB in this area and that farm-to-farm spread of disease was not an important infection mechanism. PMID- 17425935 TI - Factors associated with routine mass antimicrobial usage in fattening pig units in a high pig-density area. AB - One hundred and seven fattening pig units randomly selected in Catalonia (Spain) were surveyed during 2001-2003, in order to determine the frequency of the prophylactic use of antimicrobials and mass treatments against respiratory and enteric disease outbreaks. Logistic regression and regression tree analysis were done to determine the factors associated with the on-farm mass antimicrobial usage. Fifty-eight per cent of the surveyed farms used mass antimicrobial prophylaxis; among them, 39% used two or more antimicrobials. The more frequently administered drugs were colistin, beta-lactams and tetracyclines. Fattening units from strict finishing farms were more likely to use antimicrobials than farrow-to finish farms (Odds Ratio = 11.7) and farms with changing facilities were less likely to use these compounds (OR = 0.17). In the event of a respiratory disease outbreak, most farms applied mass treatment and 46% used two or more compounds, with tetracyclines, beta-lactams and sulphonamides being the most popular ones. Mass treatment with several compounds in case of respiratory problems was significantly related to the use of growth promoters on the farm (OR = 4.3). If an outbreak of an enteric disease occurred, most farms resorted also to a mass treatment and 48% used two or more drugs. The most frequent antimicrobial agent for the treatment of enteric disease was colistin. Fattening farms (OR = 6.9) and operations that perform routine antimicrobial prophylaxis (OR = 2.0) were more likely to use more than one antimicrobial agent for enteric diseases. PMID- 17425936 TI - Estimating the day of highly pathogenic avian influenza (H7N7) virus introduction into a poultry flock based on mortality data. AB - Despite continuing research efforts, knowledge of the transmission of the highly pathogenic avian influenza (HPAI) virus still has considerable gaps, which complicates epidemic control. The goal of this research was to develop a model to back-calculate the day HPAI virus is introduced into a flock, based on within flock mortality data. The back-calculation method was based on a stochastic SEIR (susceptible (S) - latently infected (E) - infectious (I) - removed (= dead; R)) epidemic model. The latent and infectious period were assumed to be gamma distributed. Parameter values were based on experimental H7N7 within-flock transmission data. The model was used to estimate the day of virus introduction based on a defined within-flock mortality threshold (detection rule for determining AI). Our results indicate that approximately two weeks can elapse before a noticeable increase in mortality is observed after a single introduction into a flock. For example, it takes twelve (minimum 11 - maximum 15) days before AI is detected if the detection rule is fifty dead chickens on two consecutive days in a 10 000 chicken flock (current Dutch monitoring rule for notification). The results were robust for flock size and detection rule, but sensitive to the length of the latent and infectious periods. Furthermore, assuming multiple introductions on one day will result in a shorter estimated period between infection and detection. The implications of the model outcomes for detecting and tracing outbreaks of H7N7 HPAI virus are discussed. PMID- 17425937 TI - Case-control study on feed risk factors for BSE cases born after the feed ban in France. AB - In France, after the ban on meat and bone meal (MBM) in cattle feeding in June 1990, cases of Bovine Spongiform Encephalopathy (BSE) have continued to be detected in bovines born after that ban (called BAB cases). A case-control study was therefore carried out to determine the way these cases were contaminated. A multivariate conditional model was built adjusting for the production type of the animals and taking into account the herd size. The results confirmed that feeding cattle with proprietary concentrates was at risk for BSE, with an adjusted odds ratio of 6.8 (2.5; 18.7) for the consumption of less or three different proprietary concentrates and 17.6 (5.7; 54.8) for more than three, when comparing with no consumption of proprietary concentrates, considering feeding of bovines before the age of two. The results suggest that cross-contaminations by MBM in bovine concentrates have occurred after 1990. To a lesser extent, on-farm cross contaminations, i.e. consumption by cattle of feedstuffs initially dedicated to other animals and which could legally contain MBM, have probably also existed, since the presence on farms of poultry fed purchased feed involved an increased risk of BSE with an odds ratio of 1.8 (1.1; 3.0). The use of milk replacers, which often incorporates animal fats, was also at risk with an odds ratio of 1.8 (1.0; 3.1). PMID- 17425938 TI - Molecular mechanism of liver development and regeneration. AB - The liver is the central organ for metabolism and has strong regenerative capability. Although the liver has been studied mostly biochemically and histopathologically, genetic studies using gene-targeting technology have identified a number of cytokines, intracellular signaling molecules, and transcription factors involved in liver development and regeneration. In addition, various in vitro systems such as fetal liver explant culture and primary culture of fetal liver cells have been established, and the combination of genetic and in vitro studies has accelerated investigation of liver development. Identification of the cell-surface molecules of liver progenitors has made it possible to identify and isolate liver progenitors, making the liver a unique model for stem cell biology. In this review, we summarize progresses in understanding liver development and regeneration. PMID- 17425939 TI - Molecular basis of vertebrate endoderm development. AB - The embryonic endoderm gives rise to the epithelial lining of the digestive and respiratory systems and organs such as the thyroid, lungs, liver, gallbladder, and pancreas. Studies in Xenopus, zebrafish, and mice have revealed a conserved molecular pathway controlling vertebrate endoderm development. The TGFbeta/Nodal signaling pathway is at the top of this molecular hierarchy and controls the expression of a number of key transcription factors including Mix-like homeodomain proteins, Gata zinc finger factors, Sox HMG domain proteins, and Fox forkhead factors. Here we review the function of these molecules comparing and contrasting their roles in each model organism. Finally, we will describe how our understanding of the molecular pathway governing endoderm development in embryos is being used to differentiate embryonic stem cells in vitro along endodermal lineages, with the ultimate goal of making therapeutically useful tissue. PMID- 17425940 TI - YidC as an essential and multifunctional component in membrane protein assembly. AB - Membrane proteins fulfill a number of vital functions in prokaryotic and eukaryotic cells. They are often organized in multicomponent complexes, folded within the membrane bilayer and interacting with the cytoplasmic and periplasmic or external soluble compartments. For the biogenesis of integral membrane proteins, the essential biochemical steps are (1) the insertion and topogenesis of the transmembrane protein segments into the lipid bilayer, (2) the three dimensional folding of the translocated hydrophilic domains, and (3) the assembly into multimeric complexes. Intensive research has elucidated the basic mechanisms of membrane protein insertion in the homologous translocation machineries of different cellular systems. Whereas the Sec translocation system is found in the endoplasmic reticulum of eukaryotic cells and in the prokaryotic plasma membrane, the YidC-Oxa1 membrane insertase is present in prokaryotic and organellar membranes. This review focuses on the discoveries of the YidC system in bacterial as well as the Oxa1/Alb3 protein family of eukaryotic cells and will particularly emphasize evolutionary aspects. PMID- 17425941 TI - Mitotic spindle dynamics in Drosophila. AB - Mitosis, the process by which the replicated chromosomes are segregated equally into daughter cells, has been studied for over a century. Drosophila melanogaster is an ideal organism for this research. Drosophila embryos are well suited to image mitosis, because during cycles 10-13 nuclei divide rapidly at the surface of the embryo, but mitotic cells during larval stages and spermatocytes are also used for the study of mitosis. Drosophila can be easily maintained, many mutant stocks exist, and transgenic flies expressing mutated or fluorescently labeled proteins can be made. In addition, the genome has been completed and RNA interference can be used in Drosophila tissue culture cells. Here, we review our current understanding of spindle dynamics, looking at the experiments and quantitative modeling on which it is based. Many molecular players in the Drosophila mitotic spindle are similar to those in mammalian spindles, so findings in Drosophila can be extended to other organisms. PMID- 17425943 TI - Sweet potato: a review of its past, present, and future role in human nutrition. AB - The overall objective of this chapter is to review the past, present, and future role of the sweet potato (Ipomoea batatas [L.] Lam) in human nutrition. Specifically, the chapter describes the role of the sweet potato in human diets; outlines the biochemical and nutritional composition of the sweet potato with emphasis on its beta-carotene and anthocyanin contents; highlights sweet potato utilization, and its potential as value-added products in human food systems; and demonstrates the potential of the sweet potato in the African context. Early records have indicated that the sweet potato is a staple food source for many indigenous populations in Central and South Americas, Ryukyu Island, Africa, the Caribbean, the Maori people, Hawaiians, and Papua New Guineans. Protein contents of sweet potato leaves and roots range from 4.0% to 27.0% and 1.0% to 9.0%, respectively. The sweet potato could be considered as an excellent novel source of natural health-promoting compounds, such as beta-carotene and anthocyanins, for the functional food market. Also, the high concentration of anthocyanin and beta-carotene in sweet potato, combined with the high stability of the color extract make it a promising and healthier alternative to synthetic coloring agents in food systems. Starch and flour processing from sweet potato can create new economic and employment activities for farmers and rural households, and can add nutritional value to food systems. Repositioning sweet potato production and its potential for value-added products will contribute substantially to utilizing its benefits and many uses in human food systems. Multidisciplinary, integrated research and development activities aimed at improving production, storage, postharvest and processing technologies, and quality of the sweet potato and its potential value-added products are critical issues, which should be addressed globally. PMID- 17425942 TI - Neurogenesis in the fish retina. AB - The retinas of teleost fish have long been of interest to developmental neurobiologists for their persistent plasticity during growth, life history changes, and response to injury. Because the vertebrate retina is a highly conserved tissue, the study of persistent plasticity in teleosts has provided insights into mechanisms for postembryonic retinal neurogenesis in mammals. In addition, in the past 10 years there has been an explosion in the use of teleost fish-zebrafish (Danio rerio) in particular-to understand the mechanisms of embryonic retinal neurogenesis in a model vertebrate with genetic resources. This review summarizes the key features of teleost retinal neurogenesis that make it a productive and interesting experimental system, and focuses on the contributions to our knowledge of retinal neurogenesis that uniquely required or significantly benefited from the use of a fish model system. PMID- 17425944 TI - Infectobesity: obesity of infectious origin. AB - The rapid increase in obesity and the associated health care costs have prompted a search for better approaches for its prevention and management. Such efforts may be facilitated by better understanding the etiology of obesity. Of the several etiological factors, infection, an unusual causative factor, has recently started receiving greater attention. In the last two decades, 10 adipogenic pathogens were reported, including human and nonhuman viruses, scrapie agents, bacteria, and gut microflora. Some of these pathogens are associated with human obesity, but their causative role in human obesity has not been established. This chapter presents information about the natural hosts, signs and symptoms, and pathogenesis of the adipogenic microorganisms. If relevant to humans, "Infectobesity" would be a relatively novel, yet extremely significant concept. A new perspective about the infectious etiology of obesity may stimulate additional research to assess the contribution of hitherto unknown pathogens to human obesity and possibly to prevent or treat obesity of infectious origins. PMID- 17425945 TI - Refrigerated fruit juices: quality and safety issues. AB - Fruit juices are an important source of bioactive compounds, but techniques used for their processing and subsequent storage may cause alterations in their contents so they do not provide the benefits expected by the consumer. In recent years consumers have increasingly sought so-called "fresh" products (like fresh products), stored in refrigeration. This has led the food industry to develop alternative processing technologies to produce foods with a minimum of nutritional, physicochemical, or organoleptic changes induced by the technologies themselves. Attention has also focused on evaluating the microbiological or toxicological risks that may be involved in applying these processes, and their effect on food safety, in order to obtain safe products that do not present health risks. This concept of minimal processing is currently becoming a reality with conventional technologies (mild pasteurization) and nonthermal technologies, some recently introduced (pasteurization by high hydrostatic pressure) and some perhaps with a more important role in the future (pulsed electric fields). Nevertheless, processing is not the only factor that affects the quality of these products. It is also necessary to consider the conditions for refrigerated storage and to control time and temperature. PMID- 17425946 TI - Tetrodotoxin poisoning. AB - Tetrodotoxin (TTX) is one of the most potent and oldest known neurotoxins. The poisoning cases due to ingestion of TTX-containing marine animals, especially for puffer, have frequently occurred in Asia since a long time ago. This chapter describes various topics on TTX poisoning including the tendency of poisoning incidents, typical case report, treatment and prevention, biology distribution, original source, infestation mechanism, detection methods, characteristics of chemistry and pharmacology, and therapeutic application. Furthermore, the protocols for how to make puffer safe to eat and how to prevent puffer products made from toxic puffers have been suggested. Finally, the biological significance and neurophysiological role of TTX have been elucidated and TTX may act as an important drug like anesthetic in future. PMID- 17425947 TI - Marine biotechnology for production of food ingredients. AB - The marine world represents a largely untapped reservoir of bioactive ingredients that can be applied to numerous aspects of food processing, storage, and fortification. Due to the wide range of environments they survive in, marine organisms have developed unique properties and bioactive compounds that, in some cases, are unparalleled by their terrestrial counterparts. Enzymes extracted from fish and marine microorganisms can provide numerous advantages over traditional enzymes used in food processing due to their ability to function at extremes of temperature and pH. Fish proteins such as collagens and their gelatin derivatives operate at relatively low temperatures and can be used in heat-sensitive processes such as gelling and clarifying. Polysaccharides derived from algae, including algins, carrageenans, and agar, are widely used for their ability to form gels and act as thickeners and stabilizers in a variety of foods. Besides applications in food processing, a number of marine-derived compounds, such as omega-3 polyunsaturated fatty acids and photosynthetic pigments, are important to the nutraceutical industry. These bioactive ingredients provide a myriad of health benefits, including reduction of coronary heart disease, anticarcinogenic and anti-inflammatory activity. Despite the vast possibilities for the use of marine organisms in the food industry, tools of biotechnology are required for successful cultivation and isolation of these unique bioactive compounds. In this chapter, recent developments and upcoming areas of research that utilize advances in biotechnology in the production of food ingredients from marine sources are introduced and discussed. PMID- 17425948 TI - Fruits of the actinidia genus. AB - Kiwifruit is the most well-known crop in the genus Actinidia. Although Actinidia fruit sales in the international market are dominated by a single kiwifruit cultivar Actinidia deliciosa "Hayward," there are a considerable number of cultivars and selections in the genus that have widely diverse shape, size, and hairiness. They also offer a wide variation in sensory attributes such as flesh color, flavor, and taste, and in nutritional attributes such as the vitamin C level and carotenoid content. The level of actinidin, which is a cysteine protease in kiwifruit, also varies greatly among cultivars. This chapter reviews available information related to several important components, allergenic properties, and health benefits of Actinidia fruits. PMID- 17425949 TI - Carnosine-like immunoreactivity in neurons of the brain of an advanced teleost, the gray mullet (Chelon labrosus, Risso). AB - The distribution of the dipeptide carnosine (beta-alanyl-L-histidine) and the related dipeptides anserine and homocarnosine has been studied by biochemical methods and immunohistochemistry in the brain of mammals and other tetrapods. These studies have indicated the presence of these dipeptides mainly in glial cells and in some neurons (olfactory receptor neurons and certain putative migrating neurons and neuroblasts). Here, we used immunohistochemistry with a polyclonal anti-carnosine antibody and the streptavidin-avidin method to study for the first time the distribution of carnosine/carnosine-related dipeptides in the brain of a teleost fish (the gray mullet Chelon labrosus). In order to assess the neuronal nature of carnosine-immunoreactive cells, we also used double immunofluorescence methods with antibodies to carnosine and to the neuronal protein HuC/D. The results obtained show that carnosine and/or related dipeptides are present in neurons of various brain regions. The carnosine-like immunoreactive neuronal populations of the optic tectum and cerebellum are described in detail. In the optic tectum, immunoreactivity to carnosine/carnosine related dipeptides is present in neurons of the stratum album and the stratum griseum centrale. In the cerebellum, immunoreactivity to these dipeptides is localized in Purkinje cells, in putative Golgi and stellate cells, and in many nerve fibers. Carnosine-like immunoreactive cells in mullet brain seem to be specific types of neurons, in line with previous results in a urodele but at variance with previous results in mammals, reptiles and anurans. PMID- 17425950 TI - Morphometrical and neurochemical changes in the anteroventral subdivision of the rat medial amygdala during estrous cycle. AB - The anteroventral subdivision of the medial amygdala (MeAV) is one of the vomeronasal structures involved in the control of hormonally dependent behaviors such as sexual and agonistic behaviors in rats. The present study investigates some anatomical and neurochemical parameters of this nucleus (volume, number of neurons, number of glial elements, and of NADPH-diaphorase-positive neurons) in females in two estrous cycle phases (diestrous and estrous) and in males. We also investigate the possible existence of adult neurogenesis in this nucleus in the females. Results showed that volume and estimated number of Nissl-stained neurons in the MeAV vary with the estrous cycle phase: estrous females have greater values than diestrous females. As a consequence of these variations, there is a transient sex difference between males and diestrous females. Two subpopulations of NADPH-diaphorase-positive neurons were detected: intensely stained and medium stained. The intensely stained neurons were more numerous in the estrous than the diestrous females. Neither BrdU nor GFAP inmunostaining revealed significant differences between the two groups, suggesting that adult cell generation, i.e., increases in the number of glial elements, has no significant role in the changes detected in the number of Nissl-stained sections. In conclusion, the MeAV shows functional diergism, due to plastic changes in the female rat brain probably linked to the increase of estradiol during estrous. Finally, these changes are probably functionally related to changes in the behaviors that are controlled through this nucleus. PMID- 17425951 TI - Expression of hepatocyte growth factor and c-Met after spinal cord injury in rats. AB - Since hepatocyte growth factor (HGF) plays a pivotal role in the development of the central nervous system and pathological conditions, we examined the long-term changes in the mRNA and protein expression of HGF and its receptor c-Met after spinal cord injury (SCI) in rats. HGF mRNA was significantly increased from 7 days after SCI in the injured segment, and the peak was at 7 days after SCI as assessed by real-time RT-PCR. Importantly, c-met mRNA expression was up-regulated from 1 day after SCI, and reached a peak at 14 days after SCI. Although up regulation of HGF and c-met mRNA expression in the injured segment gradually decreased, the increased expression level persisted until 56 days after SCI. Consistent with HGF mRNA expression, HGF protein level was significantly increased mainly in the injured region, which persisted until 56 days after SCI. Immunohistochemistry showed that most of GFAP-positive reactive astrocytes expressed HGF and c-Met both on 14 days and 56 days after SCI. Staining with the mitotic indicator, bromodeoxyuridine (BrdU), revealed that a small number of BrdU incorporated cells were co-localized with HGF/GFAP-positive or c-Met/GFAP positive cells both on 14 and 56 days. These data suggest that HGF and c-Met were up-regulated mainly in the reactive astrocytes around the injured region in the subacute to chronic stage of spinal cord injury. Since HGF plays a critical role in neurotrophic activity, activation of the HGF/c-Met signaling system might be involved in the process of post-traumatic regeneration. PMID- 17425952 TI - Ameliorating effect of coenzyme Q10, riboflavin and niacin in tamoxifen-treated postmenopausal breast cancer patients with special reference to lipids and lipoproteins. AB - OBJECTIVES: Tamoxifen (TAM), a non-steroidal anti-estrogen that is widely used in adjuvant therapy for all stages of breast carcinomas and in chemoprevention of high-risk group. The hepatic estrogenic effect of TAM induces hypertriglyceridemia by reduced activity of lipolytic enzymes (LPL) on triglycerides. Coenzyme Q10 (Co Q10), riboflavin and niacin are proved to be potent antioxidant and protective agents against many diseases including cancer and cardiovascular diseases (CVD). In this context, the objective of the study is to find the effect of the combined modality of Co Q10 (100 mg), riboflavin (10 mg) and niacin (50 mg) with TAM (10 mg twice a day) on serum lipids and lipoprotein levels in postmenopausal women with breast cancer. DESIGN AND METHODS: The vitamin supplementation with tamoxifen was given for a period of 90 days. Blood samples were collected at the base line, 45th and 90th day during the course of treatment. Plasma total cholesterol (TC), free cholesterol (FC), ester cholesterol (EC), phospholipids (PL), triglycerides (TGL), free fatty acids (FFA), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and very low density cholesterol (VLDL-C) were estimated in 78 untreated, only TAM-treated and combinatorialy treated group along with 46 age and sex-matched controls. RESULTS: Serum TGL and VLDL-C (p<0.001) were found to be significantly elevated and LDL-C (p<0.01), significantly reduced among TAM treated patients as compared to the untreated breast cancer subjects. All the lipids and lipoprotein levels were found to be significantly altered in the untreated breast cancer patients when compared to their normal counterparts. All the lipid and lipoprotein abnormalities were reverted back to near normal levels on 90 days of treatment on combinatorial therapy. CONCLUSION: The study figures the altered lipid and lipoprotein levels in the untreated and TAM-treated breast cancer patients. On combination therapy with Co Q10, riboflavin and niacin, it counteracts the tamoxifen-induced hyperlipidemia to normal levels. PMID- 17425953 TI - FGFs, Wnts and BMPs mediate induction of VEGFR-2 (Quek-1) expression during avian somite development. AB - Regulation of VEGFR-2 (Quek1) is an important mechanism during blood vessel formation. In the paraxial mesoderm, Quek1 expression is restricted to the lateral portion of the somite and later to sclerotomal cells surrounding the neural tube. By implanting FGF 8b/8c or SU 5402 beads into the paraxial mesoderm, we show that FGF8 in addition to BMP4 from the intermediate mesoderm (IM) is a positive regulator of VEGFR-2 (Quek1) expression in the quail embryo. The expression of Quek1 in the medial somite half is normally repressed by the notochord and Sfrps-expression in the neural tube. Over-expression of Wnt 1/3a also results in an up-regulation of Quek1 expression in the somites. We also show that up-regulation of FGF8/Wnt 1/3a leads to an increase in the number of endothelial cells, whereas inhibition of FGF and Wnt signaling by SU 5402 and Sfrp-2 results in a loss of endothelial cells. Our results demonstrate that the regulation of Quek1 expression in the somites is mediated by the cooperative actions of BMP4, FGF8 and Wnt-signaling pathways. PMID- 17425954 TI - Accelerated de novo sarcomere assembly by electric pulse stimulation in C2C12 myotubes. AB - The assembly of sarcomeres, the smallest contractile units in striated muscle, is a complex and highly coordinated process that relies on spatio-temporal organization of sarcomeric proteins, a process requiring spontaneous Ca(2+) transients. To investigate the relationship between Ca(2+) transients and sarcomere assembly in C2C12 myotubes, we employed electric pulse stimulation (EPS), which allows the frequency of Ca(2+) transients to be manipulated. We monitored contractile activity as a means of evaluating functional sarcomere establishment using the differential image subtraction (DIS) method. C2C12 myotubes initially displayed no contractility with EPS, due to a lack of sarcomere architecture. However, C2C12 myotubes showed remarkable contractile activity with EPS-induced repetitive Ca(2+) transients (1 Hz) within only 2 h. This activity was concurrent with the development of sarcomere structure. Importantly, the period required for the acquisition of contractile activity in response to excitation was dependent upon the frequency of Ca(2+) oscillations, but a sustained increase in intracellular Ca(2+) (not oscillatory) by high frequency EPS (10 Hz) was incapable of conferring either contractility or sarcomere assembly on the myotubes. The EPS-facilitated de novo functional sarcomere assembly appeared to require calpain-mediated proteolysis. In addition, modulation of integrin signals, by adding collagen IV or RGD-peptide, significantly affected the EPS-induced development of contractility. Taken together, these observations indicate that the frequency of the Ca(2+) oscillation determines the time required to establish functionally active sarcomere assembly and also suggest that the Ca(2+) oscillatory signal may be decoded through reorganization of the integrin-cytoskeletal protein complex via calpain-mediated proteolysis. PMID- 17425955 TI - Intermediate filaments: a role in epithelial polarity. AB - Intermediate filaments have long been considered mechanical components of the cell that provide resistance to deformation stress. Practical experimental problems, including insolubility, lack of good pharmacological antagonists, and the paucity of powerful genetic models have handicapped the research of other functions. In single-layered epithelial cells, keratin intermediate filaments are cortical, either apically polarized or apico-lateral. This review analyzes phenotypes of genetic manipulations of simple epithelial cell keratins in mice and Caenorhabditis elegans that strongly suggest a role of keratins in apico basal polarization and membrane traffic. Published evidence that intermediate filaments can act as scaffolds for proteins involved in membrane traffic and signaling is also discussed. Such a scaffolding function would generate a highly polarized compartment within the cytoplasm of simple epithelial cells. While in most cases mechanistic explanations for the keratin-null or overexpression phenotypes are still missing, it is hoped that investigators will be encouraged to study these as yet poorly understood functions of intermediate filaments. PMID- 17425956 TI - Anti-fibrillogenic and fibril-destabilizing activity of nicotine in vitro: implications for the prevention and therapeutics of Lewy body diseases. AB - The aggregation of alpha-synuclein (alphaS) has been implicated as a critical step in the development of Lewy body diseases (LBD) and multiple system atrophy (MSA). Both retrospective and prospective epidemiological studies have consistently demonstrated an inverse association between cigarette smoking and Parkinson's disease (PD). We used fluorescence spectroscopy with thioflavin S, electron microscopy and atomic force microscopy to examine the effects of nicotine, pyridine, and N-methylpyrrolidine on the formation of alphaS fibrils (f alphaS) from wild-type alphaS (alphaS (WT)) and A53T mutant alphaS (A53T) and on preformed f alpha Ss. Nicotine dose-dependently inhibited the f alphaS formation from both alphaS (WT) and A53T. Moreover, nicotine dose-dependently destabilized preformed f alpha Ss. These effects of nicotine were similar to those of N methylpyrrolidine. The anti-fibrillogenic activity of nicotine may be exerted not only by the inhibition of f alphaS formation but also by the destabilization of preformed f alphaS. Additionally, this effect may be attributed to N methylpyrrolidine moieties of nicotine. PMID- 17425957 TI - Acute psychosocial challenge and cardiac autonomic response in women: the role of estrogens, corticosteroids, and behavioral coping styles. AB - Theoretical statements, as well as clinical and experimental data, suggest that the amplitude of cardiovascular reactivity to acute stressors can be a good predictor of preclinical and clinical cardiovascular states. The aim of the present study is to investigate the role of estrogens, the hypothalamic-pituitary adrenocortical activity, and the behavioral profile in individual cardiac autonomic reactivity to brief laboratory stressors in women. Thirty-six adult, healthy women were exposed to a stress interview and a mental task test, each lasting 5 min. They were assigned to two experimental groups: D4, i.e. 4 days after menses beginning (follicular phase, n=18), and D14, i.e. 14 days after menses beginning (ovulatory phase, n=18). The cardiac measurements in the baseline, stress and recovery periods consisted in heart rate (average R-R interval) and parasympathetic tone (r-MSSD) quantification, while the HPA axis activity and stress reactivity were assessed via plasma cortisol and dehydroepiandrosterone concentrations. The ethological profile during the interview was drawn by means of non-verbal behavior analysis. The cardiac, adrenocortical and behavioral responses to the two stressors were similar in groups D4 and D14, despite significantly higher estradiol levels in the latter. Subjects with higher pre-stress cortisol levels had higher heart rate and lower vagal activity in the baseline, stress and recovery phases. Women showing higher level of submission were characterized by higher heart rate acceleration and vagal withdrawal during both the interview and the recovery phase. In addition, the subjects that exhibited greater displacement during the interview were also characterized by lower heart rate increments and less pronounced vagal suppression during post-stress recovery. In conclusion, the present results do not support a clear buffering role of estrogens in cardiovascular response to acute stressors. However, they confirm that baseline HPA axis activity can be predictive of cardiac autonomic activity and stress responsiveness. They also highlight the modulating role of the individual style of behavioral coping in cardiac sympathovagal stress reactivity. Therefore, the objective assessment of the individual behavioral profile via the analysis of non-verbal communication patterns might represent a powerful tool for identifying subjects with higher risk of cardiac events. PMID- 17425958 TI - Stellate ganglion block may relieve hot flashes by interrupting the sympathetic nervous system. AB - Stellate ganglion block is routinely used in pain clinics. The mechanism of action of the stellate ganglion block is uncertain; the most common explanation is that it produces peripheral vasodilation, resulting in neural inhibition in the ganglion's sphere of innervation. However, the wide range of conditions that have been reported to respond favorably to stellate ganglion block suggest that its effectiveness may not be solely the result of increased blood flow nor restricted just to its sphere of innervation. We have found that stellate ganglion block is effective in the treatment of hot flashes in postmenopausal women, as well as those with estrogen depletion resulting from breast cancer treatment. Based on evidence that hot flashes may be centrally mediated and that the stellate ganglion has links with the central nervous system nuclei that modulate body temperature, we hypothesize that the stellate ganglion block provides relief of hot flashes by interrupting the central nervous system connections with the sympathetic nervous system, allowing the body's temperature regulating mechanisms to reset. If this mechanism can be confirmed, this would provide women with intractable hot flashes with an effective, potentially long lasting means of relieving their symptoms, and potentially widen the range of indications for stellate ganglion block to include other centrally mediated syndromes. PMID- 17425959 TI - Review of collagen VII sequence variants found in Australasian patients with dystrophic epidermolysis bullosa reveals nine novel COL7A1 variants. AB - BACKGROUND: Dystrophic epidermolysis bullosa (DEB) is an inherited skin fragility disorder where blistering occurs in the sub-lamina densa zone at the level of anchoring fibrils (AFs) of the dermo-epidermal junction. Both autosomal dominant (DDEB) and recessive (RDEB) result from mutations in the type VII collagen gene (COL7A1). OBJECTIVE: The purpose of this study was to understand the genotype phenotype correlation in Australian patients with DEB. METHODS: Skin biopsies from patients were processed for immunofluorescence mapping, the COL7A1 gene was screened for sequence variants. RESULTS: We report 14 Australian families with different forms of dystrophic epidermolysis bullosa (DEB) with 23 different COL7A1 allelic variants, nine of which were novel. Four cases of RDEB-HS combined two premature termination codon (PTC) variants and three other cases of RDEB-HS with combined PTC and spice-site or glycine substitution variants. G2043R, a de novo dominant variant, was also identified in this study. Four "silent" glycine substitutions were found in this study, G2775S, G1673R, G1338V and G2719A. EB17, with combined R2791W and G2210V variants, had a DDEB-Pasini phenotype, in contrast to two family members who had severe DDEB pruriginosa, with the same genotype. CONCLUSION: In this study, the RDEB variants included nonsense variants, splice site variants, internal deletions or insertions, "silent" glycine substitutions within the triple helix or N or C terminal ends of the triple helix and non-glycine missense variants within the triple helix domain. DDEB usually involves glycine substitutions within the triple helix of COL7A1 although other missense variants or splice-site alterations may underlie some cases. PMID- 17425960 TI - Contactin1a expression is associated with oligodendrocyte differentiation and axonal regeneration in the central nervous system of zebrafish. AB - Contactin1a (Cntn1a) is a zebrafish homolog of contactin1 (F3/F11/contactin) in mammals, an immunoglobulin superfamily recognition molecule of neurons and oligodendrocytes. We describe conspicuous Cntn1a mRNA expression in oligodendrocytes in the developing optic pathway of zebrafish. In adults, this expression is only retained in glial cells in the intraretinal optic fiber layer, which contains 'loose' myelin. After optic nerve lesion, oligodendrocytes re express Cntn1a mRNA independently of the presence of regenerating axons and retinal ganglion cells upregulate Cntn1a expression to levels that are significantly higher than those during development. After spinal cord lesion, expression of Cntn1a mRNA is similarly increased in axotomized brainstem neurons and white matter glial cells in the spinal cord. In addition, reduced mRNA expression in the trigeminal/anterior lateral line ganglion in erbb3-deficient mutant larvae implies Cntn1a in Schwann cell differentiation. These complex regulation patterns suggest roles for Cntn1a in myelinating cells and neurons particularly in successful CNS regeneration. PMID- 17425961 TI - Extrapontine myelinolysis in a 4 year old with diabetic ketoacidosis. AB - Extrapontine and central pontine myelinolysis (EPM/CPM) are rare events in pediatric neurology but can have devastating consequences. They are most commonly associated with rapid correction of hyponatremia but have been reported in other situations as well. This condition is relatively more common in adult neurology, not surprisingly, as alcoholism and associated malnutrition are often predisposing conditions. There have been few case reports in children with regards to this. We describe a 4-year old who presented with focal neurological deficits in the setting of diabetic ketoacidosis and the ensuing underlying osmotic imbalances. The patient made a remarkable recovery with no deficits of note-cognitive or motor. To our knowledge this is the youngest case reported so far of EPM in a child with diabetic ketoacidosis. The history of the condition, early animal experiments, clinicopathologic correlates, previous case reports and other scenarios in which this unusual event can occur are discussed--though the exact pathogenesis of this condition still remains unclear. We hope to bring to the attention of clinicians caring for children in the acute care setting, the importance of gradual correction of serum osmolality to reduce morbidity and mortality. PMID- 17425962 TI - Proton magnetic resonance spectroscopy for radiation encephalopathy induced by radiotherapy for nasopharyngeal carcinoma. AB - OBJECTIVE: To understand the metabolic patterns of NAA, Cr and Cho in radiation encephalopathy (RE) induced by radiotherapy for nasopharyngeal carcinoma detected by proton magnetic resonance spectroscopy (1H-MRS), and provide useful evidence for early diagnosis of this disease. METHODS: Chemical shift imaging 1H-MRS was performed for 10 healthy volunteers (control group) and 21 patients with pathologically confirmed nasopharyngeal carcinomas, who developed RE after radical radiotherapy as diagnosed on the basis of clinical symptoms and imaging findings. The contents of NAA, Cr and Cho in the pixels were observed, the metabolic maps generated, and NAA/Cr and NAA/Cho ratios calculated for all the subjects. RESULTS: The concentrations of NAA, Cr and Cho were rarely observed in the necrosis and liquefaction foci in the patients, nor were any signals displayed on their metabolic maps. In the visible lesions, with the exception of the necrosis and liquefaction foci, the content of NAA increased slightly, whereas that of Cr or Cho decreased obviously or even became absent. An area around the lesion was identified where NAA content decreased but Cr or Cho increased. The signal in the metabolic maps appeared indistinguishable. The NAA/Cr and NAA/Cho ratios were less than 1. Farther away from the visible lesions, the NAA, Cr and Cho contents remained normal with NAA/Cr and NAA/Cho ratios of less than 1. CONCLUSION: 1H-MRS is capable of displaying the patterns of metabolite changes in RE induced by radiotherapy, and identifying larger area of abnormal metabolism in RE than the visible lesion in MRI, which suggests the possibility of earlier detection of RE with 1H-MRS. PMID- 17425963 TI - [Immunoediting of natural killer cells by human nasopharyngeal carcinoma cell line: altered expression of KIRs and NKG2D receptors leads to reduction of natural killer cell-mediated cytolysis]. AB - OBJECTIVE: To analyze the changes of inhibitory killer cell immunoglobulin-like receptors (KIRs), NKG2D receptor and the cytotoxicity of natural killer (NK) cells induced by persistent exposure to CNE2 cells. METHODS: The HLA-class I genotypes of CNE2 cells and KIR genotypes were determined by PCR with sequence specific primers (PCR-SSP). The expressions of KIR2DL1, KIR2DL3, KIR3DL1, and NKG2D by the NK cells (freshly isolated NK cells, NK cells cocultured with 100 U/ml IL2 or with 100 U/ml IL2 and CNE2 cells as the control, IL2 and CNE2 groups, respectively) were analyzed by flow cytometry. Cytotoxicity of NK cells against CNE2 cells were detected by LDH releasing assay. RESULTS: The HLA genotypes of CNE2 cells were A2, 24, B18, 35, Cw4, 7. NK cells isolated from 3 healthy donors expressed KIR2DL1, KIR2DL3, and KIR3DL1. After 4, 24 and 48 h of culture, NK cells in CNE2 group displayed higher KIR2DL1, KIR2DL3 but lower NKG2D expression than those in the control and IL2 groups (P<0.01), whereas the latter two groups showed no significant difference in KIR2DL1, KIR2DL3, and NKG2D expressions (P>0.05), and no difference in KIR3DL1 expression was found between the 3 groups (P>0.05). After 24 h of culture, the cytotoxicity against CNE2 cells mediated by the NK cells in IL2 and CNE2 groups were (26.96-/+1.47) % and (2.74-/+1.64) % at E:T ratios of 10:1, and (35.74-/+3.59)% and (4.57-/+2.41) % at E:T ratio of 20:1, respectively. NK cells in CNE2 group displayed lower cytotoxicity than those in IL2 group (P<0.01). CONCLUSIONS: Persistent exposure to tumor cells expressing NKG2D ligands can lead to downregulated expression of NKG2D receptor, increased expression of KIRs and reduction of NK-mediated cytolysis. These results elucidate the molecular mechanism of reduced cytotoxicity mediated by the edited NK cells and indicate that blocking HLA-class I-bound KIRs or enhancing the expression of NKG2D may promote NK cell-mediated cytolysis. PMID- 17425964 TI - [Protective effect of donator liver ischemic preconditioning on posttransplant liver graft function in Chinese miniature pigs]. AB - OBJECTIVE: To investigate the protective effect of the ischemia preconditioning of the donor liver on posttransplant liver graft function in Chinese miniature pigs. METHODS: Twenty-five partially inbred Chinese miniature pigs were randomized into three groups, namely the normal control group, the ischemia reperfusion group and ischemic preconditioning group. Biopsies of the liver graft were performed to analyze HSP70 expression by means of immunoblotting, and the changes of serum AST/ALT levels were assayed using an automated biochemical analyzer. Histopathological assessment was carried out to identify the hepatocyte injury using optical and transmission electron microscopy. RESULTS: Ischemia preconditioning resulted in a notable increase in HSP70 expression and milder injury of the hepatocyte microstructure, whereas ischemia-reperfusion caused a significant increase of serum transaminases level (P<0.01) with declined HSP70 expression and obvious microstructural changes of the liver tissue. CONCLUSION: Ischemic preconditioning can produce obvious protective effects on the donor liver, and positively regulates the expression of shock protein. PMID- 17425965 TI - [Construction of lentivirus producing PLC-gamma1 siRNA and its effect on apoptosis of human colorectal carcinomas cell lines]. AB - OBJECTIVE: To construct recombinant lentivirus that stably suppresses phospholipase C (PLC) gamma1 expression in human colorectal carcinoma LoVo cells to obtain LoVo cell lines deficient in PLC gamma1 for investigation of the role of PLC gamma1 gene. METHODS: Recombinant lentivirus producing PLC gamma1 siRNA were constructed to infect LoVo cells, and the stably transduced cells were selected with blasticidin. The protein and mRNA expression of PLC gamma1 was examined by Western blotting and RT-PCR, and the effect of the lentivirus on cell apoptosis was analyzed by flow cytometry. RESULTS AND CONCLUSION: PLC gamma1 siRNA significantly suppressed PLC gamma1 expression in LoVo cells, suggesting high efficiency of gene silencing induced by the siRNA produced by the recombinant lentivirus. Concomitantly, cell apoptosis induced by 5-FU was significantly increased. PMID- 17425966 TI - [Induced differentiation and signaling factor PTEN expression of 3T3-L1 adipocytes]. AB - OBJECTIVE: To optimize the condition for inducing the differentiation of 3T3-L1 preadipocytes into adipocytes and study the expression of PTEN tumor suppression gene in this process, aiming to understand the regulatory role of PTEN in normal adipocyte differentiation and collect laboratory evidence for developing drugs targeting PTEN. METHODS: The differentiation of 3T3-L1 preadipocytes cultured in high-glucose DMEM were induced according to 2 protocols with different combinations of dexamethasone, isobutylmethylxanthine (IBMX) and insulin, and the resultant adipocytes were identified by oil red O staining. The total proteins of 3T3-L1 were extracted and analyzed by Western blotting, and PTEN homology between mice and human was analyzed by bioinformatic method. RESULTS: For optimized 3T3 L1 differentiation, 3T3-L1 cells were initially induced with the combination of 1 micromol/L dexamethasone, 0.5 mmol/L IBMX and 5 microg/ml insulin for 48 h, followed by treatment with 5 microg/ml insulin in 4.5 g/L glucose DMEM for 48 h, which resulted in high differentiation rate of 3T3-L1 cells (up to 90% on the 10th day) with unified morphology and size. PTEN expression varied quantitatively in the process of differentiation, especially low on the 12th day as compared with those measured on days 4, 6 and 9. The mice PTEN mRNA shared 96% homology and PTEN amino acid 100% homology with their human counterparts. CONCLUSION: Endogenous PTEN expression is down-regulated during 3T3-L1 differentiation, suggesting that PTEN may enhance insulin sensitivity and promote adipogenesis under physiological conditions. PMID- 17425967 TI - [Evaluation of AB-8 macroporous adsorption resin for adsorption of total flavones in liquorice]. AB - OBJECTIVE: To evaluate the performance of AB-8 macroporous adsorption resin for adsorption and desorption of flavones in liquorice. METHODS: The concentration of flavones in liquorice was determined by ultraviolet spectrophotometry, and the adsorption behavior of AB-8 macroporous adsorption resin to flavones in liquorice was examined for the adsorption capacity and the volume of solution loaded. RESULTS: Optimal adsorption of flavones was achieved with the sample pH of 5, total flavones concentration in the solution of 0.85 mg/ml, sample flow velocity of 3 BV/h, and washing with 60% ethanol at the flow velocity of 3 BV/h. CONCLUSION: AB-8 macroporous adsorption resin can be well applicable for enrichment of flavones in liquorice. PMID- 17425968 TI - [Removal of the template molecules from vinblastine-imprinted polymer]. AB - OBJECTIVE: To investigate the optimal condition for complete removal of the template molecules from vinblastine (VLB)-imprinted polymer. METHOD: The prepared polymers were packed into the cartridges of solid-phase extraction column and washed by methanol-glacial acetic acid mixture with different proportions. The contents and recoveries of VLB in the effluents were determined. RESULTS: Polymer extraction with methanol-glacial acetic acid (9:1, V/V) resulted in VLB recovery of 91.73%, but template bleeding was observed because of incomplete VLB removal. Using methanol-glacial acetic acid (6:4, V/V) as the extraction solvent, the recovery of VLB reached 98.03% with less solvents and extract times. The polymers could selectively adsorb VLB through non-covalent interactions and still exhibited strong affinity for the template molecule but not for the structural analogue vincristine after extraction with methanol-glacial acetic acid (6:4, V/V). CONCLUSION: Methanol-glacial acetic acid (6:4, V/V) is an ideal extract solvent for complete template molecule removal from the polymers, and the processed polymers possess stable capacity of specific recognition and selectivity to the template. PMID- 17425969 TI - [Bioinformatic identification and analysis of Schistosoma japonicum adenine phosphoribosyltransferase]. AB - OBJECTIVE: To identify adenine phosphoribosyltransferases in Schistosoma japonicum and analyze their structural features. METHODS: Based on the accessible transcriptome and proteomic data, the S. japonicum adenine phosphoribosyl transferases were identified using bioinformatics approaches including bi directional homology comparison, domain search and phylogenetic analysis. Homology modeling was also performed to describe the structural features of the proteins. RESULTS AND CONCLUSION: Two homologue sequences of adenine phosphoribosyltransferase were obtained from S. japonicum, and the EST abundance, physico-chemical properties and three-dimensional structures of them were also acquired. PMID- 17425970 TI - [Effect of elastic fiber alterations in the tunica albuginea of the penis on erectile function of diabetic rats]. AB - OBJECTIVE: To investigate the effect of elastic fiber alterations in the tunica albuginea of the penis on erectile function of diabetic rats. METHODS: Streptozotocin (STZ) injection was adopted to produce rat models of diabetes mellitus and erectile dysfunction. Forty rats were randomized equally into two groups according to the time after streptozotocin (STZ) injection, namely 4 week group and 7 week group. Each group was further divided into 4 subgroups, including a control group (n=5, without STZ injection), diabetic with erectile dysfunction group (DM and ED group), diabetic without erectile dysfunction group (DM group) and group with neither diabetes mellitus or erectile dysfunction after STZ injection (None group). Victoria blue/Ponceau red staining and color image analysis were used to observe the content of the elastic fibers in the tunica albuginea, which was quantified by means of integrated optical density (IOD) readings. RESULT: Significant difference in the IOD was observed between different groups (F=10.433, P<0.001). The content of elastic fibers in the tunica albuginea was the lowest in DM and ED group among the 4 groups (P<0.05), and there was no significant difference between 7-week and 4 week groups (F=0.685, P=0.415), nor was any interaction observed (F=0.905, P=0.452). CONCLUSIONS: Decreased elastic fibers in the tunica albuginea can result from diabetes mellitus. Elastic fibers in the tunica albuginea play an important role in the course of erection, and erectile dysfunction may result from decreased elastic fiber content. PMID- 17425971 TI - [Expression of PCNA and GST-pi after stereotactic irradiation for liver cirrhosis and hepatocarcinoma in rabbits]. AB - OBJECTIVE: To observe the changes of the cytokines following stereotactic irradiation for hepatocarcinoma with cirrhosis in rabbits. METHODS: Sixteen rabbits with liver cirrhosis and hepatocarcinoma (experimental group) were randomized into two equal groups to receive stereotactic irradiation at single dose of 20 and 30 Gy, respectively. Eight rabbits with hepatocarcinoma (control group) were divided into two equal groups and treated in identical manner. All the rabbits were killed 3 weeks after irradiation, and EV two-step method was used to observe the cytokine changes of proliferating cell nuclear antigen (PCNA) and glutathione S-transferase pi (GST-pi) after irradiation. RESULTS: After irradiation, PCNA and GST-pi expression showed significant difference in the adjacent liver tissue between the experimental and control rabbits with irradiation at 20 Gy (P=0.010), but not with the irradiation dose of 30 Gy (P=1.000). Irradiation at different doses resulted in significant difference in the cytokine expression in the experimental rabbits (P=0.010). In the liver tissue exposed to irradiation, different irradiation doses resulted in significant difference in PCNA and GST-pi protein expression (P=0.010). CONCLUSIONS: For hepatocarcinoma with cirrhosis in rabbits, radiation at the single dose of 30 Gy produces better response than 20 Gy, and PCNA and GST-pi may serve as good indexes for evaluating the therapeutic effect. PMID- 17425972 TI - [Preparation of cisplatin-impregnated coral hydroxyapatite drug delivery system]. AB - OBJECTIVE: To prepare a cisplatin-impregnated coral-derived hydroxyapatite (CCHA) drug delivery system (DDS), and evaluate its inhibitory effect on human osteosarcoma cells U-2 OS, human breast cancer and prostatic carcinoma cells PC-3 in vitro. METHODS: The coral-derived hydroxyapatite (CHA) was manufactured by hydrothermal exchange and impregnated with cisplatin by vacuum freeze-drying techniques. The leaching solutions of this DDS was collected at different intervals in a course of 8 weeks and their inhibitory effect on the cells was tested in vitro by MTT assay. RESULTS: Electron microscope showed that cisplatin was distributed homogeneously in the pores of CHA. The inhibition rates of the leaching solution on all the tumor cells exceeded 50% except for PC-3 cells, whose inhibition rate was 29.92% when treated with the solution collected at the eighth week. CONCLUSION: CCHA allows sustained drug release and maintains excellent inhibitory effect on human bone tumor cells within 8 weeks in vitro. PMID- 17425973 TI - [Epidemiological application of ion chromatography for urinary iodine monitoring]. AB - OBJECTIVE: To establish a method of ion chromatography for urine iodide determination and evaluate its application in epidemiological studies. METHODS: Urinary iodine was determined using ICS-2500 ion chromatography coupled with IonPac AS7 analytical column (4 mm x 250 mm), silver electrode and direct current amperometry for 467 elderly subjects (above 50 years) with their urinary Tsh and FT4 also determined. The history of thyroid diseases was examined among these subjects. RESULTS: The peak area of the ion chromatography was linearly correlated with urinary iodine concentration, with the correlation coefficient of 0.9999. The detection limit of iodide was around 3.5 microg/L (S/N=3), with coefficients of variation ranging from 3.16% to 3.45% and mean recovery rate of 95%. The urinary iodine level in the 467 elderly subjects showed a positively skewed distribution with the median level of 224.7 microg/L. CONCLUSION: Ion chromatography for urinary iodine determination has excellent selectivity, sensitivity, reliability, accuracy and stability, and may ensure satisfactory effect for application in epidemiological studies. PMID- 17425974 TI - [Effect of 5 drugs with different antitumor mechanism on tumor-formation potential of rat hepatocarcinoma cells and mesenchymal stem cells]. AB - OBJECTIVE: To study the association of drug sensitivity of rat hepatocarcinoma cells and bone marrow mesenchymal stem cells (MSCs) to 5 antitumor drug with different antitumor mechanisms with tumorigenicity of the hepatocarcinoma cells in nude mice. METHODS: Primary liver carcinoma was induced with diethylnitrosamine in rats, and the tumor cells and MSCs were obtained from 10 of the rats. The inhibition ratio of the hepatocarcinoma cells and MSCs following treatment with the 5 drugs were measured by MTT assay. The weight of the tumor in nude mice resulting from the injection of the isolated tumor cells treated with the 5 anticancer drugs was measured 6 weeks after implantation. For each anticancer drug, the difference in the inhibition ratio of the anticancer drugs against the hepatocarcinoma cells and MSCs, and he correlation of the inhibition ratio of the anticancer drugs with implanted tumor weight were analyzed. RESULTS: No correlation was found between the inhibition ratio of the 5 anticancer drugs against the hepatocarcinoma cells and the tumor weight of nude mice, but a significant negative correlation was identified between the inhibition ratio of the MSCs and the tumor weight. CONCLUSION: MSCs have similar drug resistance mechanism to the tumor stem cells. The inhibition ratio of the anticancer drugs against the MSCs can help evaluate the invasion potential of hepatocarcinoma cells. PMID- 17425975 TI - [Changes of Smoothened expression during retinofugal pathway development in mouse embryos]. AB - OBJECTIVE: To examine the change of Smoothened (Smo) expression in the retinofugal pathway and in the growth cones during the period of embryonic day 13 (E13) to E15. METHODS: Smo expression in the chiasm and growth cones was observed by fluorescent immunostaining and retinal explant culture. RESULTS: On E13 and E14, Smo was expressed moderately in the retina and optic disc, and in the corner of the retina, Smo expression was especially dense. On E13, Smo expression was detected in the optic nerves and ventral diencephalon, but only in the superficial region of the optic tract on E14. Smo was also detected in the stem and filopodia of the growth cones in the retinal explant culture during this period. CONCLUSION: Smo expression changes in different developmental phases, suggesting that Smo might play a role in signal optic axon growth during the development of the retinofugal pathway. PMID- 17425976 TI - [Role of 5-HT2A-receptors coupled with superoxide anion in the medial area of nucleus retrofacialis]. AB - OBJECTIVE: To explore the effect of 5-HT2A-receptors coupled with superoxide anion (O2-) on respiratory regulation signal transductionin passageway in the medial area of nucleus retrofacialis (mNRF). METHODS: mNRF island was prepared from medullary slices of neonatal SD rats according to Johnson's method and transferred separately into 24-well culture plates with reagents according to protocol, followed by incubation for 60 min at 37 degrees C in a humidified incubator with 5% CO2. Absorbance of 100 microl supernatant was measured by spectrophotometry at 550 nm and the effect of 5-HT and 2,5-dimethoxy-4 iodoamphetamine hydrochloride (DOI, agonist of 5-HT2A-receptors) on O2- generation in the mNRF was observed, along with the inhibition of this effect by ketanserin (antagonist of 5-HT2A-receptors) and alpha-lipoic acid (alpha-LA, a antioxidant). RESULTS: 5-HT concentration-response curve demonstrated that absorbance peak occurred at 1 micromol/L without further increment with higher concentration. DOI concentration-response curve showed the absorbance peak at 20 micromol/L without further increment. 5-HT and DOI significantly increased the absorbance with comparable effects. Ketanserin and alpha-LA significantly decreased the absorbance generated by 5-HT and DOI. CONCLUSION: Activation of 5 HT2A receptors results in obvious O2- production in mNRF, suggesting that 5-HT2A receptors regulate respiratory function in association with O2-. PMID- 17425977 TI - [Observation of an injectable tissue-engineered bone constructed with autologous platelet-rich plasma and bone marrow stromal cells]. AB - OBJECTIVE: To construct an injectable tissue-engineered bone graft with fibrin glue (FG), autologous platelet-rich plasma (PRP) and bone marrow stromal cells (BMSCs) cultured in vitro and study its biological characteristics and microscopic structures. METHODS: BMSCs isolated from rabbit iliac bone marrow were culture-expanded in vitro. The injectable tissue-engineered bone constructed from autologous PRP, FG, and BMSCs was cultured in vitro, and its biological characteristics were observed including the time of gel formation, histological features, seed cell survival and microscopic structures. RESULTS: The constructed injectable tissue-engineered bone began gel formation within 20 to 30 s, and after a week-long culture, the gelatine began to degrade, and numerous well viable fusiform cells could be seen to adhere to the bottom of the Petri dish. Scanning electron microscopy identified globular and olivary cells embedded in the fibrin glue, and numerous small particles could be seen around of the cells. CONCLUSION: Construction of an injectable tissue-engineered bone graft with FG, BMSCs and PRP does not require sophisticated techniques and ensures good biological property of the bone graft that can be easily shaped and allow good growth of the seed cells, suggesting great potential of this technique for clinical use. PMID- 17425978 TI - [Comparison of clinical outcomes of four protocols for frozen-thawed embryo transfer cycle]. AB - OBJECTIVE: To compare the clinical outcome of 4 protocols of frozen-thawed embryo transfer cycle to select the optimal endometrial preparation method for frozen thawed embryos transfer. METHODS: A retrospective analysis of the 4 clinical protocols was conducted including natural cycle, down-regulated hormone replacement treatment (HRT) cycle, hMG cycle and natural cycle+hCG in endometrial preparation for 419 frozen-thawed embryos transfer cycle, and the clinical pregnancy rate, implantation rate, early abortion rate, ectopic pregnancy rate , ongoing pregnancy rate and delivery rate were compared between the 4 protocols. RESULTS: There was no significant difference between the 4 groups with different clinical protocols in age, duration of infertility, reason of infertility, number of embryo transferred and endometrial thickness. The 4 protocols differed little in the implantation rate, clinical pregnancy rate, biochemical pregnancy rate, early abortion rate, ectopic pregnancy rate, ongoing pregnancy rate and delivery rate in the four clinical protocols. CONCLUSION: The 4 clinical protocols for frozen-thawed embryos transfer all have favorable clinical outcome, and choice of a specific protocol should be made according to the a comprehensive consideration of the individual conditions of the patient. PMID- 17425979 TI - [Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system]. AB - OBJECTIVE: To achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli. METHODS: The gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting. RESULTS: The plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75. CONCLUSION: Efficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation. PMID- 17425980 TI - [Anatomic reduction of midfacial sag after intraoral reduction malarplasty]. AB - OBJECTIVE: To study the technical refinements of intraoral reduction malarplasty to prevent postoperative drooping of the cheek. METHODS: Twenty-four patients with prominent zygomatic complex underwent reduction malarplasty from January 2005 to January 2006. In all these cases, the osteotomized malar complex was repositioned superioposteriorly, and the perioral muscles and periosteum were redraped accordingly and fixed on deep temporal fascia. RESULTS: Symmetry between the left and right complexer were achieved, with a decreased midfacial width of 10.22-/+1.97 mm. During postoperative follow-up of 4 to 6 months, all the patients had satisfactory operative results, and reported no postoperative drooping of cheek. CONCLUSION: This method of superioposterior reduction of the perioral muscles and periosteum is simple and effective, but the long-term result needs to be observed. PMID- 17425981 TI - [Ecdysterone promotes wound healing in rabbits]. AB - OBJECTIVE: To study the efficacy of ecdysterone in promoting wound healing in rabbits. METHODS: Twenty-one New Zealand rabbits were randomized equally into 3 groups and in each rabbit, 4 separate full-thickness skin wounds were induced and treated with dexamethasone, ecdysterone and Yunnan white powder, or untreated, respectively. On day 3, 6, and 9, respectively, one group of rabbits were sacrificed and the local wound tissue was obtained for pathological examination. RESULTS: Three days after the operation, the 4 wounds in each rabbit did not show significant difference in the healing rate (P>0.05). On day 6, as was similar to day 9, ecdysterone and Yunnan white powder showed better efficacy in promoting wound healing than dexamethasone (P<0.01), the latter having similar effect with the control management (P>0.05), and that of ecdysterone was similar to Yunnan white powder (P>0.05). Pathologically, early stage of wound healing was characterized by exudation and hyperemia, and on days 6 and 9, ecdysterone and Yunnan white powder exhibited similar strong effect for shaping the granulation tissue and stimulating epithelial cell proliferation. CONCLUSION: Ecdysterone can obviously promote wound healing in rabbits, which may offer a clinical alternative for promoting wound healing. PMID- 17425982 TI - [Correlation analysis of increased blood glucose and insulin resistance after traumatic brain injury in rats]. AB - OBJECTIVE: To study the pattern of the alterations of blood glucose, insulin and insulin sensitivity after traumatic brain injury in rats, and verify the occurrence of insulin resistance after the injury. METHODS: Based on Feeney's model of brain injury, the blood glucose and insulin concentration of the dogs measured 30 min before and at 6, 12, 24, 48, 72 and 120 h after injury. BG60-120, GIR60-120, and insulin sensitivity index (ISI) reflecting the insulin sensitivity were measured at 6, 24, 48, and 72 hours following severe traumatic brain injury using euglycemic-hyperinsulinemic clamp. RESULTS: Both the blood glucose and insulin concentration increased markedly in rats following moderate and severe brain injury. BG60-120 increased markedly, and GIR60-120 and ISI decreased significantly 6, 24, 48, and 72 h after severe brain trauma as compared with those of the sham operation group. Blood glucose concentration of rats following severe injury was positively correlated with insulin concentration and BG60-120 at the corresponding time points, but negatively with GIR60-120 and ISI. CONCLUSION: Both the blood glucose and insulin concentration increase markedly in rats following severe brain injury. Increased blood glucose even in the presence of high-level insulin is due to acute insulin resistance occurring after traumatic brain injury. PMID- 17425983 TI - [Expression of liver-type fatty acid-binding protein and vascular endothelial growth factor and their correlation in human hepatocellular carcinoma]. AB - OBJECTIVE: To detect the expression of liver-type fatty acid-binding protein (L FABP) and vascular endothelial growth factor (VEGF) in human hepatocellular carcinoma (HCC) and its adjacent liver tissues, and investigate the correlation between the expressions of L-FABP and VEGF and their role in the occurrence and progression of HCC. METHODS: Reverse transcriptase-polymerase chain reaction (RT PCR) and immunohistochemical (IHC) staining were employed to examine the expression of L-FABP and VEGF in HCC and its adjacent liver tissues obtained from the surgical specimens of 61 HCC patients who underwent liver resections in West China hospital. RESULTS: The results of RT-PCR showed that the expression level of L-FABP and VEGF in HCC was significantly higher than that in its adjacent liver tissues (L-FABP: 0.97-/+0.12, 0.83-/+0.14, t=5.21, P<0.05; VEGF: 0.92 /+0.11, 0.59-/+0.15, t=11.79, P<0.05). L-FABP tended to co-express with VEGF (P<0.05). IHC staining revealed that the expression of L-FABP and VEGF was mainly located in the cytoplasm, and the gray scale of L-FABP expression was significantly higher than that in the adjacent liver tissues (92.73-/+7.67, 82.83 /+6.90, t=7.44, P<0.05). The number of L-FABP- and VEGF-positive cells in HCC was significantly lower than that in the adjacent liver tissues (L-FABP: 92.18 /+4.44, 84.52-/+6.43, t=5.94, P<0.05; VEGF: 88.69-/+5.56, 77.64-/+5.93, t=8.72, P<0.05). Co-expression of L-FABP and VEGF observed in RT-PCR and also in IHC (P<0.05). CONCLUSION: Both L-FABP and VEGF expressions are up-regulated in HCC. L FABP gene may be involved in the carcinogenesis of human HCC. Expression of L FABP is associated with VEGF expression, suggesting that L-FABP promotes the growth of blood vessels by taking up the fatty acids from the bloodstream, and both of them produce a marked effect on energy metabolism in HCC. PMID- 17425984 TI - [A new Bayesian reconstruction algorithm for PET images based on correction of the detected sinogram data]. AB - OBJECTIVE: To improve Bayesian reconstruction of positron-emission tomography (PET) images by devising a novel coupled feedback (CF) iterative model. METHODS: The CF iterative algorithm was applied to update the noisy detected emission sinogram data using the latest reconstructed image in the iterative process of PET reconstruction. The relevant operations included linear filtering, wiener filtering, and projection of the reconstructed images. The sinogram data used in the study was obtained from simulated phantom data. RESULTS: The experiments and corresponding visional and quantitative comparisons showed that the new method had better performance than the traditional Bayesian reconstruction of PET images for improvement of the PET images. CONCLUSIONS: The proposed sinogram-correcting method allows improvement on the original measurement data, and is applicable for PET image reconstruction or other reconstruction tasks with high noise level. PMID- 17425985 TI - [Inhibitory effects of reduced glutathione sodium on renal nuclear factor-kappaB expression in rats with diabetes of different stages]. AB - OBJECTIVE: To investigate the relation between diabetic nephropathy and nuclear factor-kappaB (NF-kappaB) expression, and observe the effect of reduced glutathione sodium (GSH) on NF-kappaB activation and in prevention of diabetic nephropathy. METHODS: Seventy male Sprague-Dawley rats weighing 200-/+25 g were randomized into control group (10 rats) and diabetic group (60 rats, subgrouped into 1 month, 3 months, 6 months and their corresponding intervention subgroups, each consisting of 10 rats). The rats in the 6 diabetic groups were subjected to intraperitoneal injection of streptozotocin, and those in the control group received injection with 0.1 mmol/L citric acid buffer solution of the same volume. The diabetic models were affirmed upon a fasting blood glucose >or=16.5 mmol/L 3 days after the injection. The intervention groups were injected intraperitoneally with GSH (10 mg/100 g) once daily. Fasting blood glucose and body weight were measured every week. The rats were executed at the end of 1, 3, and 6 months respectively and the nucleoproteins were extracted from the renal specimen. NF-kappaB was measured using electrophoretic mobility shift assay (EMSA) after labeling with isotope probe, and the gray scale of the electrophoretic bands was analyzed. RESULTS: EMSA optical density analysis of electrophoretic bands showed that NF-kappaB expression increased in each diabetic groups in comparison with the control group (P<0.05), and NF-kappaB level rose proportionally with the disease course of 1 month, 3 months and 6 months. The activity of NF-kappaB decreased in the intervention groups as compared with the corresponding untreated groups (P<0.05). CONCLUSION: The activation of NF-kappaB plays a role in the onset and development of diabetes. NF-kappaB inhibition and containment of inflammation might be one of the mechanisms of GSH treatment for diabetes. PMID- 17425986 TI - [Clinical features and prognosis of IgA nephropathy with glomerular crescents]. AB - OBJECTIVE: To understand the clinical features and prognosis of IgA nephropathy (IgAN) patients with glomerular crescents. METHODS: The clinical data collected at the time of renal biopsy and the follow-up data of 89 IgAN patients with glomerular crescents were analyzed with 412 IgAN patients without crescents serving as the control group. Follow-up study was conducted in 78 patients with crescents and 198 without it, and the renal survival rate was estimated using Kaplan-Meier survival analysis. RESULTS: The incidence of glomerular crescents was 17.8% in IgAN patients. Clinically, 39 patients with crescents experienced rapidly progressive glomerulonephritis, resulting in a significantly higher rate of this manifestation than that in patients without crescent. Patients with crescents also had more grave pathological changes in the glomerulus and renal tubule interstitium than the control patients. Patients were followed up for an average of 40.3-/+29.6 months in crescent group and 45.1-/+26.9 months in the control group, and the 1-, 3-, 5-year renal survival rate was 95.24%, 80.95%, 61.9% in the former and 100%, 91.67%, 79.17% in the latter, respectively. CONCLUSION: IgAN patients with crescents have severer clinical and pathological manifestations and poorer prognosis than those without crescents. PMID- 17425987 TI - [Effects of different interventions for early-stage acute lung injury on respiratory response and work of breathing in canine model]. AB - OBJECTIVE: To evaluate the effects of different interventional strategies, namely controlled high-concentration oxygen therapy, continuous positive airway pressure (CPAP) and bi-level positive airway pressure (BiPAP) ventilation, on respiratory response and work of breathing (WOB) in canine models of early-stage acute lung injury (ALI). METHODS: After successful duplication of ALI models with oleic acid (diagnostic criteria: Pa(O2)/Fi(O2)0.05), which maintained the level of early ALI/ARDS stage. BiPAP greatly improved V(T) and V(T)/Ti, showing better effects than CPAP and O2. No significant differences were noted among the 3 groups in T(I)/T(tot) (P>0.05). BiPAP showed superior effect to CPAP in lowering the peak transdiaphragmatic pressure (Pdi). CPAP and BiPAP both effectively counteracted intrinsic positive end expiratory pressure (PEEPi) (P<0.01), while O2 produced no obvious such effects (P>0.05). BiPAP showed the most evident effects, followed by CPAP, in reducing WOB, but oxygen therapy produced no obvious effects. CPAP (P<0.01) and BiPAP (P>0.05) both effectively reduced the proportion of ingredients in WOB related to PEEPi. CONCLUSION: BiPAP and CPAP can produce favorable effects in relieving dyspnea, reducing WOB and improving respiratory response to control the deterioration of ARDS. BiPAP has more significant therapeutic effects than CPAP and oxygen therapy. PMID- 17425988 TI - [Effects of oxymatrine on expression of nuclear factor kappa B in kidneys of rats with adriamycin-induced chronic renal fibrosis]. AB - OBJECTIVE: To investigate the effects of oxymatrine on the expression of nuclear factor kappa B (NF-kappaB) in the kidneys of rats with adriamycin-induced chronic renal fibrosis. METHODS: Totally 120 Wistar rats were randomly assigned to normal control group, renal fibrosis model group, benazepril treatment group and oxymatrine treatment group (n=30). The rats in the normal control were injected with normal saline via the tail vein, and those in the other 3 groups with adriamycin (2 mg/kg) on the 7th day and 21st day of the experiment, respectively. Oxymatrine (100 mg/kg) or benazepril (6 mg/kg) was given by gastric perfusion after the second injection. Every 4 weeks after the second injection, 5 rats in each group were killed to evaluate the pathological changes and functional impairment of the kidney. Immunohistochemistry was used to detect the expression of NF-kappaB and inhibitory kappa B (IkappaB) in the kidney. The association of NF-kappaB expression with IkappaB expression, renal pathological changes and functional impairment were studied. RESULTS: Oxymatrine and benazepril ameliorated renal fibrosis and functional impairment. Immunohistochemical staining revealed increased NF-kappaB expression and decreased IkappaB expression in the model group in comparison with oxymatrine and benazepril treatment groups 8 weeks after the second injection, but no significant difference was noted between the latter two groups. NF-kappaB expression in the kidneys of rats with adriamycin-induced chronic renal fibrosis showed an inverse correlation with IkappaB expression and positive correlation with pathological changes and functional impairment. CONCLUSION: Oxymatrine may inhibit renal fibrosis by down regulating NF-kappaB expression, which may play a key role in protection against renal fibrosis. PMID- 17425989 TI - [Cloning of tight junction protein claudin-1 and construction of the mammalian expression vector]. AB - OBJECTIVE: To construct a recombinant plasmid containing the coding region of tight junction protein claudin-1 gene to understand the functional role of claudin-1 in human colorectal carcinoma. METHODS: The total RNA was extracted using Trizol from human colorectal carcinoma cell line SW620, and the DNA for claudin-1 was obtained by means of RT-PCR. The PCR product was inserted into the plasmid pEGFP-C1 after restriction endonuclease digestion and ligation. The recombinant plasmid was then transfected into human colorectal carcinoma cell line SW480. RESULTS: The sequence of the recombinant plasmid was verified by restriction endonuclease and DNA sequence analysis, and the target protein expression was detected mostly on the cell membrane. CONCLUSION: The expression vector claudin-1/pEGFP-C1 has been constructed successfully and the target protein can be expressed in human colorectal carcinoma cell line. PMID- 17425990 TI - [MRI findings of uterine cervical cancer and value of MRI in preoperative staging]. AB - OBJECTIVE: To evaluate the value of magnetic resonance imaging (MRI) in diagnosis and preoperative staging of uterine cervical cancer. METHODS: MRI findings and staging in 72 patients with cervical carcinoma were retrospectively analyzed, and the size, location, signal intensity and invasion of the tumor were observed. MRI sequence included SE T1WI, (TSE)T2WI, T2WI/SPIR and contrast-enhanced T1WI. RESULTS: MRI identified uterus cervical cancer in all cases with the exception of only 1 case of IA stage. The tumor was represented by hypointensity and isointensity on T1WI, heterogeneous and homogeneous hyperintensity on T2WI, mildly heterogeneous enhancement after bolus intravenous GD-DTPA injection. MRI had an accuracy of 86% in localization of the tumor, but its accuracy in clinical staging was only 64% (chi2=6.453, P<0.05). The tumor volume measured by MRI was similar with that by pathological measurement (1.94-/+1.15 vs 1.94-/+1.11, P>0.05). CONCLUSION: MRI can accurately describe the size and invasion of uterine cervical cancer, especially useful in detecting parametrial invasion, but for diagnosis of IA uterine cervical cancer, MRI findings are not sufficient without considerations of clinical findings and cellular examination. PMID- 17425991 TI - [Clinical study of double filtration plasmapheresis for treatment of refractory myasthenia gravis]. AB - OBJECTIVE: To assess the therapeutic effect, timing of administration, complication prevention and management of the double filtration plasmapheresis (DFP) in the treatment of refractory myasthenia gravis (MG). METHODS: Thirty-one patients with refractory MG were treated with KM 8800 membrane plasmapheresis monitor. DFP was performed every 3 days and the exchanging liquid was composed of 50 ml of 20% albumen and 1000 ml plasma substitute. Physical examination for absolute clinical score and blood sample was collected for AchR-Ab determination early in the morning on days 0, 3, 7, 14 of DFP. RESULTS: With a total effective rate of 91.9%, complete recovery, basic recovery, improvement, and response was achieved in 2, 4, 11, and 17 patients, respectively, whereas the other 3 failed to respond. Hypotension occurred twice in 2 cases and was corrected after symptomatic treatment. CONCLUSION: DFP may effectively lower blood AchR-Ab level of with minimal complications, and can be valuable for treatment of refractory MG. PMID- 17425992 TI - [Effects of arsenic trioxide on apoptosis of peripheral T-lymphocytes from asthmatic patients and normal subjects in vitro]. AB - OBJECTIVE: To study the effects of arsenic trioxide on apoptosis of peripheral T lymphocytes from asthmatic patients and normal subjects in vitro. METHODS: The T lymphocytes were isolated from the blood of 21 asthmatic patients and 20 healthy controls and treated with arsenic trioxide and dexamethasone. Cell apoptosis was observed by fluorescence microscope and measured with flow cytometry and Cytochrome C ELISA kit. RESULTS: The T-lymphocytes from the asthmatic patients, when compared to those from of the healthy control, exhibited decelerated spontaneous apoptosis after a 24-hour incubation in vitro. Dexamethasone treatment significantly increased the percentage of apoptotic T-lymphocytes from both asthmatic patients and normal subjects in comparable magnitude. Arsenic trioxide treatment, in contrast, significantly increased the percentage of apoptotic T-lymphocytes from asthmatic patients, but slightly affected the cells from the control group. CONCLUSIONS: Spontaneous apoptosis of T-lymphocytes can be decelerated in asthmatic patients, whose T-lymphocytes are more sensitive to arsenic trioxide-induced apoptosis than those of normal subjects, but the T lymphocytes from normal subjects and asthmatic patients are equally sensitive to dexamethasone. PMID- 17425993 TI - [Pituitary down-regulation and ovulation stimulation with gradually increased gonadotropin in controlled ovarian hyperstimulation]. AB - OBJECTIVE: To evaluate the efficacy of ovulation stimulation protocol with gradual increment of gonadotropin in women with high ovarian response. METHODS: A retrospective study was conducted between june 2005 and April 2006 in 70 women undergoing controlled ovarian hyperstimulation. The clinical outcomes of the women using gradual increment protocol were compared with those of women receiving other ovulation-stimulating protocols. RESULTS: The mean number of large follicles (>or=14 mm) and retrieved oocytes on the day of retrival was significantly lower, but the duration of stimulation was significantly longer in the gradual increment group than in the control group. The rate of follicular puncture was also higher in the former group. The clinical pregnancy rate, total gonadotropin dosage, cancellation rate and incidence of ovarian hyperstimulation syndrome were similar for the two groups. CONCLUSION: Ovulation stimulation protocol with gradually increased gonadotropin may provide a promising alternative for controlled ovarian hyperstimulation in women with a strong ovarian response. PMID- 17425994 TI - [Vitamin D receptor gene polymorphism is not associated with bone mineral density of pre-menopausal women in Guangzhou]. AB - OBJECTIVE: To investigate the prevalence of vitamin D receptor (VDR) gene polymorphism in pre-menopausal women in Guangzhou and study its relationship with bone mineral density(BMD). METHODS: The genotypes of VDR gene in 193 per menopausal women in Guangzhou were analyzed by polymerase chain reaction restriction fragment length polymorphism. BMD of the lumbar spine, femoral neck, greater trochanter and Ward's triangle were measured by dual-energy X-ray absorptiometry. RESULTS: In the 193 subjects, 120 (66.2%) were identified as VDR bb genotype, 64 (33.2%) as Bb, and 9 (4.6%) as BB. The b allele frequencies reached 78.76%, and B allele frequencies was 21.24%. The distribution followed the Hardy-Weinberg equilibrium. No significant difference was found in BMD among the subjects with different genotypes. CONCLUSION: VDR genotype is not related to BMD, and VDR polymorphism can not be used as a genetic marker for predicting the risk of osteoporosis in pre-menopausal women in Guangzhou. PMID- 17425995 TI - [Changes of heme oxygenase-1 and inflammatory factors in patients with end-stage renal failure following hemodialysis]. AB - OBJECTIVE: To evaluate the changes in heme oxygenase-1 (HO-1), C-reactive protein (CRP), interleukin-6 (IL-6), and malondialdehyde (MDA) levels in patients with end-stage renal failure (ESRF) following hemodialysis. METHODS: The urea nitrogen and the blood creatinine levels were determined with an automatic biochemistry analyzer, HO-1 and IL-6 levels by enzyme linked immunosorbent assay (ELISA), CRP by chemoluminescence technique, and MDA by thiobarbituric acid assay. RESULTS: HO 1, CRP, IL-6, and MDA levels were significantly elevated in patients with ESRF in comparison with the control group. After hemodialysis, HO-1 and CRP levels increased and IL-6 and MDA levels decreased significantly. CONCLUSION: In ESRF patients who have elevated inflammatory factors and HO-1 levels, hemodialysis can increase HO-1 and CRP levels and lower IL-6 and MDA levels. PMID- 17425996 TI - [Treatment of high-energy tibia plateau injury with half-ring external fixation combined with minimum internal fixation]. AB - OBJECTIVE: To evaluate the effect of less invasive surgical treatment for high energy tibia plateau injury with half-ring external fixation combined with minimum internal fixation. METHODS: From January, 2003 to May,2006, 16 cases of high-energy tibia plateau fracture were treated with half-ring external fixation combined with minimum internal fixation including 10 cases of type V and 6 cases of type VI according to Schatzker's classification. The average age of the patients was 42.4 years (range 25 to 50 years). RESULTS: All patients were followed for an average of 16 months (range 5 to 27 months). All the fractures healed after an average time of 3.5 months. Two patients developed infections of the pin holes. According to the criteria of Rasmussen, excellent results were achieved in 10, good results in 4 cases, moderate in 2, and poor in none. CONCLUSION: Half-ring external fixation combined with minimum internal fixation can be ideal for treatment of complex tibia plateau fracture. PMID- 17425997 TI - [Radiosensitivity of nasopharyngeal carcinoma in relation to cell cycle synchronization effect of tumor necrosis factor alpha]. AB - OBJECTIVE: To investigate the effect of tumor necrosis factor alpha (TNFalpha) on radiosensitivity of nasopharyngeal carcinoma (NPC) in relation to TNFalpha induced cell cycle synchronization. METHODS: The radio-resistance of a NPC cell line subclone CNE-2Z-S1 was verified by in vivo experiments and flow cytometry was performed to evaluate cell cycle synchronization in TNFalpha-treated CNE-2Z S1 cells. The radiosensitivity of the cell synchronized CNE-2Z-S1 cells was determined by clone formation in vitro and in vivo experiment in nude mice. RESULTS: TNFalpha was capable of inducing cell cycle arrest and synchronization of CNE-2Z-S1 cells. Pretreatment with TNFalpha remarkably enhanced the radiosensitivity of CNE-2Z-S1 in vitro, and in vivo experiments with nude mice also suggested the role of TNFalpha in enhancing the radiosensitivity of NPC. CONCLUSION: TNFalpha can enhance the radiosensitivity of NPC cells by inducing cell cycle synchronization. PMID- 17425998 TI - [Growth of craniopharyngioma involving the third ventricular floor in relation to the hypothalamus]. AB - OBJECTIVE: To investigate the growth of craniopharyngioma involving the third ventricular floor with regard to the hypothalamus by detecting expressions of leukocyte common antigen (CD45) and intercellular adhesion molecule (ICAM-1) in the tumor tissue. METHODS: The expressions of CD45 and ICAM-1 proteins in 30 craniopharyngioma samples with third ventricular floor involvement were detected by SP immunohistochemistry. RESULTS: The inflammations labeled by CD45 were identified commonly in the craniopharyngioma tissues involving the third ventricular floor. The expression of ICAM-1 was mainly in the inner tumor cells and interstitial cells, but not detected in the basilar tumor cells growing toward the third ventricular floor. Adamantinomatous craniopharyngiomas showed markedly higher CD45 and ICAM-1 expressions than squamous papillary tumors (P<0.05). CONCLUSION: Inflammatory adhesion largely characterizes the growth of the craniopharyngioma tissues involving the third ventricular floor toward the hypothalamus without the tendency of invasion. The difference in the inflammation between the two types of craniopharyngioma may affect the prognosis of the patients. PMID- 17425999 TI - [Relation between embryo transfer distance from the fundus and the pregnancy rate]. AB - OBJECTIVE: To investigate the association of embryo transfer distance from the fundus (TDF) with the clinical pregnancy rate (PR). METHODS: Between January 2005 and September 2006, ultrasound (US)-guided embryo transfer was conducted in 610 women. TDF was measured by transabdominal US according to which the subjects were divided into 4 groups. The association between TDF and PR or between TDF and abnormal pregnancy was analyzed. RESULTS: No significant differences in PR were observed among those 4 groups, but PR was the highest in the group with TDF >or=2 cm, and the lowest in the group 1 cm. The rate of early spontaneous abortion was higher in the group with TDF >or=2 cm. CONCLUSION: TDF variation does not produce significant difference in PR, but the distance of 1.0-2.0 cm is recommended. PMID- 17426000 TI - [Percutaneous intradiscal oxygen-ozone injection for lumbar disc herniation: no need of perioperative antibiotic prophylaxis]. AB - OBJECTIVE: To evaluate the feasibility of no antibiotic administration to prevent infection during the perioperative period of percutaneous intradiscal ozone injection for treatment of lumbar disc herniation. METHODS: Seventy-two patients with lumbar disc herniation but normal body temperature as well as normal results of three routine tests (blood, urine, stool) and C-reactive protein (CRP) level were randomly divided into two groups. The patients in prophylaxis group were given cephalothin V(2.0 g) intravenous 30 min before the operation, and the control group did not use any antibiotics. All the patients were injected with 6 10 ml ozone (40 microg/ml) for medical use into the discs with 21G needles under fluoroscopic guidance, followed by 10 ml ozone into the paravertebral space. Three days later the general examinations and CRP measurement were repeated. RESULTS: No infection was found in these patients, nor were any significant differences noted in the results of the examinations between the two groups after controlling in patients with above-normal white blood cell count, neutrophil percentage and CRP level. CONCLUSION: Prophylaxis antibiotics is not necessary during the perioperative period of percutaneous intradiscal ozone injection for lumbar disc herniation. PMID- 17426001 TI - [Comparison of three analgesic methods for postoperative pain relief and their effects on plasma interleukin-6 concentration following radical surgery for gastric carcinoma]. AB - OBJECTIVE: To compare the efficacy of preemptive epidural analgesia combined with postoperative epidural analgesia, postoperative epidural analgesia alone and intravenous analgesia for postoperative pain relief and their effects on plasma interleukin-6 (IL-6) concentration following radical surgery for gastric carcinoma. METHODS: Sixty-six patients with gastric carcinoma scheduled for gastrectomy were randomly divided into 3 groups, namely group P (n=22), group E (n=22) and group V (n=22), to receive preemptive epidural analgesia combined with postoperative epidural analgesia, exclusive postoperative epidural analgesia, and exclusive postoperative intravenous analgesia, respectively. Hemodynamic data were recorded for all the patients during the operation, and visual analogue scale (VAS) was used to assess the pain intensity at 4, 8, 16, 24, 48 and 72 h after surgery. Plasma IL-6 concentration was determined before surgery and at 24, 48, 72 h after surgery. RESULTS: No significant changes occurred in the hemodynamics during the preoperative periods. VAS and IL-6 were lower in group P than in group E and V, and group E had lower measurement than group V (P<0.05). CONCLUSION: Preemptive epidural analgesia combined with postoperative epidural analgesia provides more satisfactory pain relief and more effectively prevents IL 6 increment than exclusive epidural analgesia or intravenous analgesia after gastrectomy for gastric carcinoma. PMID- 17426002 TI - [Therapeutic efficacy of strontium-89-chloride for bone metastatic tumors without bone pain]. AB - OBJECTIVE: To evaluate the safety and efficacy of strontium-89-chloride for management of bone metastases in patients without bone pain. METHODS: Fifty-four patients without painful bone metastases were given a single intravenous dose (1.48-2.22 MBq/kg) of strontium-89-chloride, which was repeated once or twice at the interval between 3 and 6 months. RESULTS: The total response rate was 74.0% in these, and the response rate was significantly lower in patients with focal size>2 cm than in those with focal size10-fold) mRNA levels of FKS2, the alternate catalytic subunit of glucan synthase, partially restoring glucan synthase activity. Consistent with these results, up-regulation of PKC-Slt2 signaling and overexpression of FKS1 or FKS2 alleviated sensitivity of pgs1Delta to cell wall-perturbing agents and restored growth at elevated temperature. These findings demonstrate that functional Pgs1p is essential for cell wall biogenesis and activation of the PKC-Slt2 signaling pathway. PMID- 17426022 TI - Epitope analysis of the malaria surface antigen pfs48/45 identifies a subdomain that elicits transmission blocking antibodies. AB - Pfs48/45, a member of a Plasmodium-specific protein family, displays conformation dependent epitopes and is an important target for malaria transmission-blocking (TB) immunity. To design a recombinant Pfs48/45-based TB vaccine, we analyzed the conformational TB epitopes of Pfs48/45. The Pfs48/45 protein was found to consist of a C-terminal six-cysteine module recognized by anti-epitope I antibodies, a middle four-cysteine module recognized by anti-epitopes IIb and III, and an N terminal module recognized by anti-epitope V antibodies. Refolding assays identified that a fragment of 10 cysteines (10C), comprising the middle four cysteine and the C-terminal six-cysteine modules, possesses superior refolding capacity. The refolded and partially purified 10C conformer elicited antibodies in mice that targeted at least two of the TB epitopes (I and III). The induced antibodies could block the fertilization of Plasmodium falciparum gametes in vivo in a concentration-dependent manner. Our results provide important insight into the structural organization of the Pfs48/45 protein and experimental support for a Pfs48/45-based subunit vaccine. PMID- 17426023 TI - Structural and functional analysis of phi29 p16.7C dimerization mutants: identification of a novel aromatic cage dimerization motif. AB - Prokaryotic DNA replication is compartmentalized at the cellular membrane. The Bacillus subtilis phage varphi29-encoded membrane protein p16.7 is one of the few proteins known to be involved in the organization of prokaryotic membrane associated DNA replication. The functional DNA binding domain of p16.7 is constituted by its C-terminal half, p16.7C, which forms high affinity dimers in solution and which can form higher order oligomers. Recently, the solution and crystal structures of p16.7C and the crystal structure of the p16.7C-DNA complex have been solved. Here, we have studied the p16.7C dimerization process and the structural and functional roles of p16.7 residues Trp-116 and Asn-120 and its last nine C-terminal amino acids, which form an extended tail. The results obtained show that transition of folded dimers into unfolded monomers occurs without stable intermediates and that both Trp-116 and the C-terminal tail are important for dimerization and functionality of p16.7C. Residue Trp-116 is involved in formation of a novel aromatic cage dimerization motif, which we call "Pro cage." Finally, whereas residue Asn-120 plays a minor role in p16.7C dimerization, we show that it is critical for both oligomerization and DNA binding, providing further evidence that DNA binding and oligomerization of p16.7C are coupled processes. PMID- 17426024 TI - The role of a single-stranded nucleotide loop in transcriptional regulation of the human sod2 gene. AB - Manganese superoxide dismutase (MnSOD), a mitochondrial antioxidant enzyme, is necessary for survival of aerobic life. Previously, we demonstrated that a Sp1 based promoter is essential for constitutive transcription and a NF-kappaB-based intronic enhancer is responsible for cytokine-mediated induction. Here we show that nucleophosmin (NPM), a RNA-binding protein, binds to an 11G single-stranded loop in the promoter region and serves to integrate the Sp1 and NF-kappaB responses. Disruption of the loop structure causes a reduction of both constitutive and inductive transcription due to loss of the binding motif for NPM. Interaction of NF-kappaB.NPM.Sp1 facilitated by binding of NPM to the loop structure in the promoter region appears to comprise the basic complex for the transcriptional stimulation. These results suggest a novel molecular mechanism for communication between the enhancer and the GC-rich promoter. PMID- 17426025 TI - The microneme proteins EtMIC4 and EtMIC5 of Eimeria tenella form a novel, ultra high molecular mass protein complex that binds target host cells. AB - Eimeria tenella, in common with other parasitic protozoa of the phylum Apicomplexa, invades host cells using an actinomyosin-powered "glideosome" complex and requires the secretion of adhesive proteins from the microneme organelles onto the parasite surface. Microneme proteins of E. tenella include EtMIC4, a transmembrane protein that has multiple thrombospondin type I domains and calcium-binding epidermal growth factor-like domains in its extracellular domain, and EtMIC5, a soluble protein composed of 11 tandemly repeated domains that belong to the plasminogen-apple-nematode superfamily. We show here that EtMIC4 and EtMIC5 interact to form an oligomeric, ultrahigh molecular mass protein complex. The complex was purified from lysed parasites by non-denaturing techniques, and the stoichiometry was shown to be [EtMIC4](2):[EtMIC5](1), with an octamer of EtMIC4 bound non-covalently to a tetramer of EtMIC5. The complex is formed within the parasite secretory pathway and is maintained after secretion onto the surface of the parasite. The purified complex binds to a number of epithelial cell lines in culture. Identification and characterization of this complex contributes to an overall understanding of the role of multimolecular protein complexes in specific interactions between pathogens and their hosts during infection. PMID- 17426026 TI - Molecular characterization of the Ran-binding zinc finger domain of Nup153. AB - The nuclear pore complex is the gateway for selective traffic between the nucleus and cytoplasm. To learn how building blocks of the pore can create specific docking sites for transport receptors and regulatory factors, we have studied a zinc finger module present in multiple copies within the nuclear pores of higher eukaryotes. All four zinc fingers of human Nup153 were found to bind the small GTPase Ran with dissociation constants ranging between 5 and 40 mum. In addition a fragment of Nup153 encompassing the four tandem zinc fingers was found to bind Ran with similar affinity. NMR structural studies revealed that a representative Nup153 zinc finger adopts the same zinc ribbon structure as the previously characterized Npl4 NZF module. Ran binding was mediated by a three-amino acid motif (Leu(13)/Val(14)/Asn(25)) located within the two zinc coordination loops. Nup153 ZnFs bound GDP and GTP forms of Ran with similar affinities, indicating that this interaction is not influenced by a nucleotide-dependent conformational switch. Taken together, these studies elucidate the Ran-binding interface on Nup153 and, more broadly, provide insight into the versatility of this zinc finger binding module. PMID- 17426027 TI - Escherichia coli MutS tetramerization domain structure reveals that stable dimers but not tetramers are essential for DNA mismatch repair in vivo. AB - The Escherichia coli mispair-binding protein MutS forms dimers and tetramers in vitro, although the functional form in vivo is under debate. Here we demonstrate that the MutS tetramer is extended in solution using small angle x-ray scattering and the crystal structure of the C-terminal 34 amino acids of MutS containing the tetramer-forming domain fused to maltose-binding protein (MBP). Wild-type C terminal MBP fusions formed tetramers and could bind MutS and MutS-MutL-DNA complexes. In contrast, D835R and R840E mutations predicted to disrupt tetrameric interactions only allowed dimerization of MBP. A chromosomal MutS truncation mutation eliminating the dimerization/tetramerization domain eliminated mismatch repair, whereas the tetramer-disrupting MutS D835R and R840E mutations only modestly affected MutS function. These results demonstrate that dimerization but not tetramerization of the MutS C terminus is essential for mismatch repair. PMID- 17426028 TI - Asp-120 locates Zn2 for optimal metallo-beta-lactamase activity. AB - Metallo-beta-lactamases are zinc-dependent hydrolases that inactivate beta-lactam antibiotics, rendering bacteria resistant to them. Asp-120 is fully conserved in all metallo-beta-lactamases and is central to catalysis. Several roles have been proposed for Asp-120, but so far there is no agreed consensus. We generated four site-specifically substituted variants of the enzyme BcII from Bacillus cereus as follows: D120N, D120E, D120Q, and D120S. Replacement of Asp-120 by other residues with very different metal ligating capabilities severely impairs the lactamase activity without abolishing metal binding to the mutated site. A kinetic study of these mutants indicates that Asp-120 is not the proton donor, nor does it play an essential role in nucleophilic activation. Spectroscopic and crystallographic analysis of D120S BcII, the least active mutant bearing the weakest metal ligand in the series, reveals that this enzyme is able to accommodate a dinuclear center and that perturbations in the active site are limited to the Zn2 site. It is proposed that the role of Asp-120 is to act as a strong Zn2 ligand, locating this ion optimally for substrate binding, stabilization of the development of a partial negative charge in the beta-lactam nitrogen, and protonation of this atom by a zinc-bound water molecule. PMID- 17426029 TI - Beta-dystroglycan as a target for MMP-9, in response to enhanced neuronal activity. AB - Matrix metalloproteinase-9 has recently emerged as an important molecule in control of extracellular proteolysis in the synaptic plasticity. However, no synaptic targets for its enzymatic activity had been identified before. In this report, we show that beta-dystroglycan comprises such a neuronal activity-driven target for matrix metalloproteinase-9. This notion is based on the following observations. (i) Recombinant, autoactivating matrix metalloproteinase-9 produces limited proteolytic cleavage of beta-dystroglycan. (ii) In neuronal cultures, beta-dystroglycan proteolysis occurs in response to stimulation with either glutamate or bicuculline and is blocked by tissue inhibitor of metalloproteinases 1, a metalloproteinase inhibitor. (iii) Beta-dystroglycan degradation is also observed in the hippocampus in vivo in response to seizures but not in the matrix metalloproteinase-9 knock-out mice. (iv) Beta-dystroglycan cleavage correlates in time with increased matrix metalloproteinase-9 activity. (v) Finally, beta dystroglycan and matrix metalloproteinase-9 colocalize in postsynaptic elements in the hippocampus. In conclusion, our data identify the beta-dystroglycan as a first matrix metalloproteinase-9 substrate digested in response to enhanced synaptic activity. This demonstration may help to understand the possible role of both proteins in neuronal functions, especially in synaptic plasticity, learning, and memory. PMID- 17426030 TI - Selective inhibition of the collagenase activity of cathepsin K. AB - Cathepsin K, the main bone degrading protease, and chondroitin 4-sulfate (C4-S) form a complex with enhanced collagenase activity. In this report, we demonstrate the specific inhibition of the collagenase activity of cathepsin K by negatively charged polymers without affecting the overall proteolytic activity of the protease. Three different mechanisms to interfere with cathepsin-catalyzed collagen degradation are discussed: 1) inhibition of the formation of the cathepsin K/C4-S complex, 2) inhibition of the attachment of C4-S to collagen, and 3) masking of the collagenase cleavage sites in collagen. By targeting these interaction sites, collagen degradation can be modulated while the non collagenolytic activities of cathepsin K remain intact. The main inhibitory effect on collagen degradation is due to the impeding effect on the active cathepsin K/C4-S complex. Essential structural elements in the inhibitor molecules are negative charges which compete with the sulfate groups of C4-S in the cathepsin K/C4-S complex. The inhibitory effect can be controlled by length and charge of the polymers. Longer negatively charged polymers (e.g. polyglutamates, oligonucleotides) tend to inhibit all three mechanisms, whereas shorter ones preferentially affect the cathepsin K/C4-S complex. PMID- 17426031 TI - The decorin sequence SYIRIADTNIT binds collagen type I. AB - Decorin belongs to the small leucine-rich repeat proteoglycan family, interacts with fibrillar collagens, and regulates the assembly, structure, and biomechanical properties of connective tissues. The decorin-collagen type I binding region is located in leucine-rich repeats 5-6. Site-directed mutagenesis of this 54-residue-long collagen-binding sequence identifies Arg-207 and Asp-210 in leucine-rich repeat 6 as crucial for the binding to collagen. The synthetic peptide SYIRIADTNIT, which includes Arg-207 and Asp-210, inhibits the binding of full-length recombinant decorin to collagen in vitro. These collagen-binding amino acids are exposed on the exterior of the beta-sheet-loop structure of the leucine-rich repeat. This resembles the location of interacting residues in other leucine-rich repeat proteins. PMID- 17426032 TI - Arabidopsis thaliana squalene epoxidase 1 is essential for root and seed development. AB - Squalene epoxidase converts squalene into oxidosqualene, the precursor of all known angiosperm cyclic triterpenoids, which include membrane sterols, brassinosteroid phytohormones, and non-steroidal triterpenoids. In this work, we have identified six putative Arabidopsis squalene epoxidase (SQE) enzymes and used heterologous expression in yeast to demonstrate that three of these enzymes, SQE1, SQE2, and SQE3, can epoxidize squalene. We isolated and characterized Arabidopsis sqe1 mutants and discovered severe developmental defects, including reduced root and hypocotyl elongation. Adult sqe1-3 and sqe1-4 plants have diminished stature and produce inviable seeds. The sqe1-3 mutant accumulates squalene, consistent with a block in the triterpenoid biosynthetic pathway. Therefore, SQE1 function is necessary for normal plant development, and the five SQE-like genes remaining in this mutant are not fully redundant with SQE1. PMID- 17426033 TI - Glyoxylate and pyruvate are antagonistic effectors of the Escherichia coli IclR transcriptional regulator. AB - The Escherichia coli isocitrate lyase regulator (IclR) regulates the expression of the glyoxylate bypass operon (aceBAK). Founding member of a large family of common fold transcriptional regulators, IclR comprises a DNA binding domain that interacts with the operator sequence and a C-terminal domain (C-IclR) that binds a hitherto unknown small molecule. We screened a chemical library of more than 150 metabolic scaffolds using a high-throughput protein stability assay to identify molecules that bind IclR and then tested the active compounds in in vitro assays of operator binding. Glyoxylate and pyruvate, identified by this method, bound the C-IclR domain with KD values of 0.9+/-0.2 and 156.2+/-7.9 microM, as defined by isothermal titration calorimetry. Both compounds altered IclR interactions with operator DNA in electrophoretic mobility shift assays but showed an antagonistic effect. Glyoxylate disrupted the formation of the IclR/operator complex in vitro by favoring the inactive dimeric state of the protein, whereas pyruvate increased the binding of IclR to the aceBAK promoter by stabilizing the active tetrameric form of the protein. Structures of the C-IclR domain alone and in complex with each effector were determined at 2.3 A, confirming the binding of both molecules in the effector recognition site previously characterized for the other representative of the family, the E. coli AllR regulator. Site-directed mutagenesis demonstrated the importance of hydrophobic patch formed by Met-146, Leu-154, Leu-220, and Leu-143 in interactions with effector molecules. In general, our strategy of combining chemical screens with functional assays and structural studies has uncovered two small molecules with antagonistic effects that regulate the IclR-dependent transcription of the aceBAK operon. PMID- 17426034 TI - The role of complex formation between the Escherichia coli hydrogenase accessory factors HypB and SlyD. AB - The Escherichia coli protein SlyD is a member of the FK-506-binding protein family of peptidylprolyl isomerases. In addition to its peptidylprolyl isomerase domain, SlyD is composed of a molecular chaperone domain and a C-terminal tail rich in potential metal-binding residues. SlyD interacts with the [NiFe] hydrogenase accessory protein HypB and contributes to nickel insertion during biosynthesis of the hydrogenase metallocenter. This study examines the HypB-SlyD complex and its significance in hydrogenase activation. Protein variants were prepared to delineate the interface between HypB and SlyD. Complex formation requires the HypB linker region located between the high affinity N-terminal Ni(II) site and the GTPase domain of the protein. In the case of SlyD, the deletion of a short loop in the chaperone domain abrogates the interaction with HypB. Mutations in either protein that disrupt complex formation in vitro also result in deficient hydrogenase production in vivo, indicating that the contact between HypB and SlyD is important for hydrogenase maturation. Surprisingly, SlyD stimulates release of nickel from the high affinity Ni(II)-binding site of HypB, an activity that is also disrupted by mutations that affect complex formation. Furthermore, a SlyD truncation lacking the C-terminal metal-binding tail still interacts with HypB but is deficient in stimulating metal release and is not functional in vivo. These results suggest that SlyD could activate metal release from HypB during metallation of the [NiFe] hydrogenase. PMID- 17426035 TI - NMR investigation of Tyr105 mutants in TEM-1 beta-lactamase: dynamics are correlated with function. AB - The existence of coupled residue motions on various time scales in enzymes is now well accepted, and their detailed characterization has become an essential element in understanding the role of dynamics in catalysis. To this day, a handful of enzyme systems has been shown to rely on essential residue motions for catalysis, but the generality of such phenomena remains to be elucidated. Using NMR spectroscopy, we investigated the electronic and dynamic effects of several mutations at position 105 in TEM-1 beta-lactamase, an enzyme responsible for antibiotic resistance. Even in absence of substrate, our results show that the number and magnitude of short and long range effects on (1)H-(15)N chemical shifts are correlated with the catalytic efficiencies of the various Y105X mutants investigated. In addition, (15)N relaxation experiments on mutant Y105D show that several active-site residues of TEM-1 display significantly altered motions on both picosecond-nanosecond and microsecond-millisecond time scales despite many being far away from the site of mutation. The altered motions among various active-site residues in mutant Y105D may account for the observed decrease in catalytic efficiency, therefore suggesting that short and long range residue motions could play an important catalytic role in TEM-1 beta-lactamase. These results support previous observations suggesting that internal motions play a role in promoting protein function. PMID- 17426036 TI - Certain pairs of ubiquitin-conjugating enzymes (E2s) and ubiquitin-protein ligases (E3s) synthesize nondegradable forked ubiquitin chains containing all possible isopeptide linkages. AB - It is generally assumed that a specific ubiquitin ligase (E3) links protein substrates to polyubiquitin chains containing a single type of isopeptide linkage, and that chains composed of linkages through Lys(48), but not through Lys(63), target proteins for proteasomal degradation. However, when we carried out a systematic analysis of the types of ubiquitin (Ub) chains formed by different purified E3s and Ub-conjugating enzymes (E2s), we found, using Ub mutants and mass spectrometry, that the U-box E3, CHIP, and Ring finger E3s, MuRF1 and Mdm2, with the E2, UbcH5, form a novel type of Ub chain that contains all seven possible linkages, but predominantly Lys(48), Lys(63), and Lys(11) linkages. Also, these heterogeneous chains contain forks (bifurcations), where two Ub molecules are linked to the adjacent lysines at Lys(6) + Lys(11), Lys(27) + Lys(29), or Lys(29) + Lys(33) on the preceding Ub molecule. However, the HECT domain E3s, E6AP and Nedd4, with the same E2, UbcH5, form homogeneous chains exclusively, either Lys(48) chains (E6AP) or Lys(63) chains (Nedd4). Furthermore, with other families of E2s, CHIP and MuRF1 synthesize homogeneous Ub chains on the substrates. Using the dimeric E2, UbcH13/Uev1a, they attach Lys(63) chains, but with UbcH1 (E2-25K), MuRF1 synthesizes Lys(48) chains on the substrate. We then compared the capacity of the forked heterogeneous chains and homogeneous chains to support proteasomal degradation. When troponin I was linked by MuRF1 to a Lys(48)-Ub chain or, surprisingly, to a Lys(63)-Ub chain, troponin I was degraded rapidly by pure 26S proteasomes. However, when linked to the mixed forked chains, troponin I was degraded quite poorly, and its polyUb chain, especially the forked linkages, was disassembled slowly by proteasome-associated isopeptidases. Because these Ring finger and U-box E3s with UbcH5 target proteins for degradation in vivo, but Lys(63) chains do not, cells probably contain additional factors that prevent formation of such nondegradable Ub-conjugates and that protect proteins linked to Lys(63)-Ub chains from proteasomal degradation. PMID- 17426037 TI - Activation of the kinase activity of ATM by retinoic acid is required for CREB dependent differentiation of neuroblastoma cells. AB - The ATM protein kinase is mutated in ataxia telangiectasia, a genetic disease characterized by defective DNA repair, neurodegeneration, and growth factor signaling defects. The activity of ATM kinase is activated by DNA damage, and this activation is required for cells to survive genotoxic events. In addition to this well characterized role in DNA repair, we now demonstrate a novel role for ATM in the retinoic acid (RA)-induced differentiation of SH-SY5Y neuroblastoma cells into post-mitotic, neuronal-like cells. RA rapidly activates the activity of ATM kinase, leading to the ATM-dependent phosphorylation of the CREB protein, extrusion of neuritic processes, and differentiation of SH-SY5Y cells into neuronal-like cells. When ATM protein expression was suppressed by short hairpin RNA, the ATM-dependent phosphorylation of CREB was blocked. Furthermore, ATM negative cells failed to differentiate into neuronal-like cells when exposed to retinoic acid; instead, they underwent cell death. Expression of a constitutively active CREBVP16 construct, or exposure to forskolin to induce CREB phosphorylation, rescued ATM negative cells and restored differentiation. Furthermore, when dominant negative CREB proteins with mutations in either the CREB phosphorylation site (CREBS133A) or the DNA binding domain (KCREB) were introduced into SH-SY5Y cells, retinoic acid-induced differentiation was blocked and the cells underwent cell death. The results demonstrate that ATM is required for the retinoic acid-induced differentiation of SH-SY5Y cells through the ATM dependent-phosphorylation of serine 133 of CREB. These results therefore define a novel mechanism for activation of the activity of ATM kinase by RA, and implicate ATM in the regulation of CREB function during RA-induced differentiation. PMID- 17426038 TI - Differences in prognostic factors and survival among White men and Black men with prostate cancer, California, 1995-2004. AB - The authors conducted a study to determine whether differences in prostate cancer survival between White men and Black men are reduced or eliminated after accounting for differences in prognostic factors. Using population-based statewide cancer registry data, the authors analyzed data from a cohort of 122,375 non-Hispanic White men and Black men from California who were newly diagnosed with prostate cancer between 1995 and 2004 and followed through 2004. Compared with White men, Black men were characterized by younger age at diagnosis, more distant stage, less treatment with surgery or radiation therapy, higher tumor grades, lower neighborhood socioeconomic status, and more recent year of diagnosis. Adjusted only for age, the hazard ratio for prostate cancer death (Blacks vs. Whites) was 1.61 (95% confidence interval (CI): 1.50, 1.72). Additional adjustment for potentially modifiable factors (stage and treatment) eliminated most of the racial difference in survival (adjusted hazard ratio = 1.10, 95% CI: 1.03, 1.18). The racial difference in survival was completely eliminated after further adjustment for other factors (grade, socioeconomic status, and year of diagnosis) (adjusted hazard ratio = 0.99, 95% CI: 0.92, 1.06). Thus, the large difference in prostate cancer survival between White men and Black men was completely explained by known prognostic factors, with potentially modifiable disparities playing the largest role. PMID- 17426039 TI - Smoking cigarettes before first childbirth and risk of breast cancer. AB - Inconsistent epidemiologic findings on cigarette smoking and female breast cancer risk may reflect insufficient assessment of smoking onset and amount relative to reproductive events. To determine the risk of breast cancer associated with smoking during different periods of reproductive life, the authors evaluated 906 incident breast cancer cases in a nationwide cohort of 56,042 female US radiologic technologists (1983-1998) who responded to two questionnaire surveys. After they accounted for age, birth cohort, and established breast cancer risk factors, smoking-related breast cancer risks differed by smoking during three reproductive time periods (p = 0.003), with a statistically significant 3% increase per pack-year of smoking between menarche and first childbirth (relative risk = 1.03, 95% confidence interval: 1.02, 1.05) and no significant association for smoking after first childbirth. Risk also increased with younger age at smoking initiation (p-trend = 0.06), after adjustment for pack-years of smoking before and after first childbirth, indicating an independent effect of age at smoking initiation. The findings from this study suggest that sensitivity of the female breast to tobacco carcinogens is increased during adolescence and early adulthood but decreases after first childbirth, when most breast tissue has terminally differentiated. PMID- 17426040 TI - Using multiple cause-of-death data to investigate associations and causality between conditions listed on the death certificate. AB - Death rarely results from only one cause, and it can be caused by a variety of factors. Multiple cause-of-death data files can list as many as 20 contributing causes of death in addition to the reported underlying cause of death. Analysis of multiple cause-of-death data can provide information on associations between causes of death, revealing common combinations of events or conditions which lead to death. Additionally, physicians report the causal train of events through which they believe that different conditions or events may have led to each other and ultimately caused death. In this paper, the authors discuss methods used in studying associations between reported causes of death and in investigating commonly reported causal pathways between events or conditions listed on the death certificate. PMID- 17426041 TI - Re: "Bias in clinical intervention research". PMID- 17426042 TI - Comparison of adolescents with and without diabetes on indices of psychosocial functioning for three years. AB - OBJECTIVE: To determine whether diabetes is associated with psychosocial difficulties over the transition to adolescence. METHODS: We compared adolescents with diabetes (n = 132) with a healthy comparison group (n = 131) on indices of psychosocial functioning for 3 years. We interviewed both groups annually and had one parent complete a questionnaire. RESULTS: There were no group differences in depressive symptoms, anxiety, anger, or behavioral problems. However, adolescents with diabetes showed greater declines in social acceptance compared with healthy adolescents, and a greater rise in disturbed eating behavior. Over time, depressive symptoms and anxiety increased and self-worth decreased for females but not males; however, these differences were not qualified by group CONCLUSIONS: Diabetes is not associated with indicators of psychological distress from early to middle adolescence, but may be associated with the emergence of social difficulties and eating disturbances. Gender differences in psychological distress emerged, replicating past research. PMID- 17426043 TI - Lessons in pediatric neuropsycho-oncology: what we have learned since Johnny Gunther. AB - OBJECTIVE: This article provides a commentary on the progression of research over the last six decades into the neurobehavioral outcomes of children treated for intracranial tumors. METHODS: Published studies and literature reviews are surveyed, with illustrations from the author's ongoing longitudinal study. RESULTS: Research on late effects in pediatric brain tumors continues to increase in both volume and quality. Samples are larger and more differentiated, scope of outcome measurement has increased, and more powerful developmental research designs are being used. Particularly promising recent developments are described, including research on: imaging-behavior relationships, improved modeling of dose volume heterogeneity in radiation therapy, improved late effects measurement, and treatments for neurobehavioral sequelae. CONCLUSIONS: There is now a large body of scientific evidence of increasing sophistication regarding the outcomes of patients receiving the most toxic treatments. It is argued that more research into the neuropsychological effects associated with "benign" tumors of childhood is needed. PMID- 17426044 TI - Brief report: psychosocial functioning of fathers as primary caregivers of pediatric oncology patients. AB - OBJECTIVE: To evaluate the psychosocial functioning of fathers as primary caregivers of pediatric oncology patients. METHODS: Fathers who identified themselves as the primary medical caregivers were given a packet of questionnaires, including the Brief Symptom Inventory (BSI), the Impact of Event Scale (IES), the Impact on Family Scale (IFS), the Caregiver Strain Questionnaire (CGSQ), and the Parent Experience of Child Illness (PECI) scale, to complete and return by mail. The 23 fathers who returned the questionnaire packets were compared with 23 mothers who were matched on demographic variables. RESULTS: There were no differences between groups on self-report measures of distress or illness-related parenting stress. Descriptively, however, the majority of parents were above normative means on measures of psychological distress with a significantly greater proportion of fathers endorsing elevated levels of depression on the BSI. CONCLUSION: Including fathers in pediatric psychosocial research is important and represents a growing trend in psycho-oncology. PMID- 17426045 TI - The role of parental monitoring in adolescent health outcomes: impact on regimen adherence in youth with type 1 diabetes. AB - OBJECTIVE: To determine if parental monitoring of adolescent behavior was related to regimen adherence and metabolic control among adolescents with type 1 diabetes. An additional objective was to compare the relative importance of instrumental parenting behaviors such as monitoring to affective behaviors such as parental support as predictors of regimen adherence. METHOD: Ninety-nine adolescents aged 12-18 years and their primary caregiver completed self-report questionnaires. Path analysis was used to test a model where diabetes-specific parental monitoring and support were predicted to have direct effects on regimen adherence and indirect effects on metabolic control via regimen adherence and an alternative model where parental support moderated the effects of monitoring on adherence. RESULTS: Diabetes-specific, but not general, monitoring was found to be associated with regimen adherence based on both parent and youth report. Monitoring had an indirect effect on metabolic control through regimen adherence. Although adolescent-reported parental support was significantly associated with regimen adherence in bivariate analyses, multivariate analyses indicated that parental support was not a significant independent predictor of health outcomes when parental monitoring was considered simultaneously. Modest support was also found for parental support as a moderator of the relationship between monitoring and adherence. CONCLUSIONS: Close parental monitoring of care completion can contribute to better adherence in adolescents with diabetes. General warmth and support in the absence of careful parental supervision may be insufficient to help youth achieve adequate levels of adherence. PMID- 17426046 TI - Using radon risk to motivate smoking reduction II: randomized evaluation of brief telephone counseling and a targeted video. AB - Radon and cigarette smoking have synergistic effects on lung cancer risk. Electric utility company bill stuffers offered free radon test kits to households with at least one smoker. Participating households (n = 1364) were randomized within a 2 x 2 design to evaluate the main effects of brief telephone counseling and a targeted video on smoking cessation and the establishment of new household smoking bans. Phone counseling was associated with cessation at 3-month follow-up but neither intervention led to 12-month or sustained cessation. While neither intervention had a significant effect on new bans, there were trends in the predicted direction and the combination of the two significantly increased new bans compared with no intervention. The presence of children in the household was associated with new bans. While few households had high levels of radon, such levels were associated with radon mitigation behaviors. Together with a previous study, these results suggest radon risk is a useful and inexpensive way to engage smoking households in risk reduction behaviors, especially the institution of household smoking bans. PMID- 17426047 TI - Sex selectivity of mouse ultrasonic songs. AB - In many species, reproduction requires detecting, recognizing, and courting a potential mate. Progress through these stages is guided by cues involving a wide range of sensory systems. Here we explore the tasks of detection, recognition, and response in terms of the ultrasonic songs of male mice presented with odor cues contained in urine. We find that the quantity of singing, more so than specific features of the songs, varies depending upon the odor cue. For experienced male mice, responses to female odor cues depend only on the concentration of female cues and are independent of the presence of male cues. However, for naive mice, male cues appear to be synergistic for the response to female cues. We therefore find no direct behavioral evidence for a role of opponent neural processing, such as lateral inhibition, in distinguishing sex by olfactory cues. However, modeling demonstrates that lateral inhibition could be one possible mechanism to account for the switch from synergy to independence. PMID- 17426048 TI - Odor-sampling time of mice under different conditions. AB - Response accuracy and odor sample times on positive (S+) and negative (S-) trials were recorded for mice trained on a variety of go, no-go odor detection and discrimination tasks. Odor sample time was relatively stable over extended training on the same task, increased during acquisition of difficult tasks, relatively insensitive to reinforcement magnitude, and, in some cases, provided more information regarding task difficulty and discrimination than did response accuracy. Mice generally sampled longer on S- trials in simple odor detection tasks but longer on S+ trials in odor discrimination tasks. PMID- 17426049 TI - Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carassius auratus) exposed to a glyphosate formulation using the micronucleus test and the comet assay. AB - Glyphosate is a widely used broad-spectrum weed control agent. In the present study, an in vivo study on the genotoxic effects of a technical herbicide (Roundup) containing isopropylamine salt of glyphosate was carried out on freshwater goldfish Carassius auratus. The fish were exposed to three doses of glyphosate formulation (5, 10 and 15 ppm). Cyclophosphamide at a single dose of 5 mg/l was used as positive control. Analysis of micronuclei, nuclear abnormalities and DNA damage were performed on peripheral erythrocytes sampled at intervals of 48, 96 and 144 h posttreatment. Our results revealed significant dose-dependent increases in the frequencies of micronuclei, nuclear abnormalities as well as DNA strand breaks. Our findings also confirmed that the alkaline comet assay and nuclear deformations in addition to micronucleus test on fish erythrocytes in vivo are useful tools in determining the potential genotoxicity of commercial herbicides. PMID- 17426050 TI - Dimerization of the Rad50 protein is independent of the conserved hook domain. AB - The Mre11 complex (Mre11-Rad50-Nbs1) is involved in a diverse array of DNA metabolic processes including the response to DNA double-strand breaks (DSBs). The structure of Rad50 plays a key role in the DNA-binding and end-bridging activity of the complex. An interesting feature within the central portion of the Rad50 protein is the Rad50 hook region that is defined by the highly conserved CXXC motif. The structure of the Pyrococcus furiosus Rad50 hook region revealed an intermolecular dimerization of Rad50 through the coordination of a zinc ion by the four cysteines. Biochemical and genetic analysis in Saccharomyces cerevisiae have shown that mutations in the conserved cysteines impact all functions of the Mre11 complex including interaction with Mre11, increased sensitivity to DSB inducing agents, telomere maintenance and intrachromosomal association. Mutations in the yeast hook domain can lead to increased chromosome fragmentation, suggesting that the hook domain of Rad50 is essential for the tethering of chromosome ends. In this study, we have examined the effects of mutating the key cysteine residues in the hook domain of human Rad50 (hRad50), focusing on the interactions Rad50 has with itself, Mre11 and DNA. Our results reveal that mutation of the conserved cysteine residues abrogates dimerization at the hook domain in hRad50; however, disrupting dimerization at this domain does not appear to impair the interaction of full-length hRad50 with itself and hMre11 or affect DNA-binding activity of the hMre11-Rad50 complex. PMID- 17426051 TI - Schizophrenia and toxoplasmosis. PMID- 17426052 TI - MAP17 overexpression is a common characteristic of carcinomas. AB - We undertook a large-scale genetic screen to identify genes able to alter the cellular response to physiological signals and provide selective advantage once tumorigenesis has begun. We identified MAP17, a small 17 kDa non-glycosylated membrane protein previously identified, being overexpressed in carcinomas. We found that MAP17 is overexpressed in a great variety of human carcinomas. Immunohistochemical analysis of MAP17 during cancer progression shows, at least in prostate and ovarian carcinomas, that overexpression of the protein strongly correlates with tumoral progression (P < 0.0001). Many tumor cells also express MAP17 and its expression does not correlate with expression of SCL, a neighbor gene reported to be co-expressed in some hematopoietic cell lines. SCL neither is expressed in most MAP17-positive tumors, indicating the independent transcription of MAP17, at least in carcinomas. We cloned 5' genomic region to MAP17 and described the minimal promoter necessary to produce independent activation of MAP17. Moreover, we have found that MAP17 promoter is activated by oncogenes. Taken together, our data show an independent activation of MAP17 promoter that can be driven by oncogenes and that might explain the common overexpression of MAP17 in human carcinomas. PMID- 17426053 TI - Will pay-for-performance and quality reporting affect health care disparities? AB - Pay-for-performance (P4P) and public quality-reporting programs can increase the quality of health care for the services being measured. However, unless carefully designed, these programs may have the unintended consequence of increasing racial and ethnic disparities. This paper describes ways in which P4P and public reporting programs may increase disparities and suggests ways in which programs might be designed that will make them likely to reduce, or at least not increase, disparities. PMID- 17426054 TI - Proteome analysis of grape skins during ripening. AB - The characterization of proteins isolated from skin tissue is apparently an essential parameter for understanding grape ripening as this tissue contains the key compounds for wine quality. It has been particularly difficult to extract proteins from skins for analysis by two-dimensional electrophoresis gels and, therefore, a protocol for this purpose has been adapted. The focus was on the evolution of the proteome profile of grape skin during maturation. Proteome maps obtained at three stages of ripening were compared to assess the extent to which protein distribution differs in grape skin during ripening. The comparative analysis shows that numerous soluble skin proteins evolve during ripening and reveal specific distributions at different stages. Proteins involved in photosynthesis, carbohydrate metabolisms, and stress response are identified as being over-expressed at the beginning of colour-change. The end of colour-change is characterized by the over-expression of proteins involved in anthocyanin synthesis and, at harvest, the dominant proteins are involved in defence mechanisms. In particular, increases in the abundance of different chitinase and beta-1,3-glucanase isoforms were found as the berry ripens. This observation can be correlated with the increase of the activities of both of these enzymes during skin ripening. The differences observed in proteome maps clearly show that significant metabolic changes occur in grape skin during this crucial phase of ripening. This comparative analysis provides more detailed characterization of the fruit ripening process. PMID- 17426055 TI - The ectopically parting cells 1-2 (epc1-2) mutant exhibits an exaggerated response to abscisic acid. AB - The ECTOPICALLY PARTING CELLS 1 (EPC1) gene encodes a putative retaining glycosyltransferase of the GT64 family, and epc1-1 mutant plants have a severely dwarfed phenotype. A new mutant allele of this gene, epc1-2, has been isolated. Reduced cell adhesion that has previously been reported for the epc1-1 mutant was not observed for either the epc1-1 or epc1-2 mutants grown in our conditions, suggesting that EPC1 does not affect cell adhesion but is involved in some other process affecting plant growth and development. It is shown that the epc1-2 mutant exhibits hypersensitivity to the phytohormone abscisic acid in germination and root elongation assays, however it shows an unaltered response to gibberellin, epi-brassinosteroid, auxin, or ethylene. An EPC1:YFP fusion protein is localized to small motile structures within the cytosol that are similar in size and number to the Golgi apparatus. Analysis of cell wall pectins revealed that levels of beta-(1,4)-galactan in the epc1-2 mutant are reduced by 50%, whilst other pectic polysaccharides (homogalacturonan, arabinan, and rhamnogalacturonan II) are unchanged. PMID- 17426056 TI - Ultraviolet A-specific induction of anthocyanin biosynthesis in the swollen hypocotyls of turnip (Brassica rapa). AB - Ultraviolet A (UV-A)-mediated regulation of anthocyanin biosynthesis was investigated in swollen hypocotyls of the red turnip 'Tsuda'. The shaded swollen hypocotyls which contained negligible anthocyanin were exposed to artificial light sources including low fluence UV-B, UV-A, blue, red, far-red, red plus UV A, far-red plus UV-A, and blue plus red. Among these lights, only UV-A induced anthocyanin biosynthesis and co-irradiation of red or far-red with UV-A did not affect the extent of UV-A-induced anthocyanin accumulation. The expression of phenylalanine ammonia lyase (PAL; EC 4.3.1.5), chalcone synthase (CHS; EC 2.3.1.74), flavanone 3-hydroxylase (F3H; EC 1.14.11.9), dihydroflavonol 4 reductase (DFR; EC 1.1.1.219), and anthocyanidin synthase (ANS; EC 1.14.11.19) genes was increased with time during a 24 h exposure to UV-A. In contrast, irradiation with red, blue, UV-B, and a combination of blue with red failed to induce CHS expression. Microarray analysis showed that only a few genes, including CHS and F3H, were induced significantly by UV-A, while a separate set of many genes was induced by low fluence UV-B. The UV-A-specific induction of anthocyanin biosynthesis and the unique gene expression profile upon UV-A irradiation as compared with blue and UV-B demonstrated that the observed induction of anthocyanin biosynthesis in red turnips was mediated by a distinct UV-A-specific photoreceptor, but not by phytochromes, UV-A/blue photoreceptors, or UV-B photoreceptors. PMID- 17426057 TI - Phytotoxic effects of trichothecenes on the growth and morphology of Arabidopsis thaliana. AB - Non-volatile sesquiterpenoids, a trichothecene family of phytotoxins such as deoxynivalenol (DON) and T-2 toxin, contain numerous molecular species and are synthesized by phytopathogenic Fusarium species. Although trichothecene chemotypes might play a role in the virulence of individual Fusarium strains, the phytotoxic action of individual trichothecenes has not been systematically studied. To perform a comparative analysis of the phytotoxic action of representative trichothecenes, the growth and morphology of Arabidopsis thaliana growing on media containing these compounds was investigated. Both DON and diacetoxyscirpenol (DAS) preferentially inhibited root elongation. DON-treated roots were less organized compared with control roots. Moreover, preferential inhibition of root growth by DON was also observed in wheat plants. In addition, T-2 toxin-treated seedlings exhibited dwarfism with aberrant morphological changes (e.g. petiole shortening, curled dark-green leaves, and reduced cell size). These results imply that the phytotoxic action of trichothecenes differed among their molecular species. Cycloheximide (CHX)-treated seedlings displayed neither feature, although it is known that trichothecenes inhibit translation in eukaryotic ribosomes. Microarray analyses suggested that T-2 toxin caused a defence response, the inactivation of brassinosteroid (BR), and the generation of reactive oxygen species in Arabidopsis. This observation is in agreement with our previous reports in which trichothecenes such as T-2 toxin have an elicitor-like activity when infiltrated into the leaves of Arabidopsis. Since it has been reported that BR plays an important role in a broad range of disease resistance in tobacco and rice, inactivation of BR might affect pathogenicity during the infection of host plants by trichothecene-producing fungi. PMID- 17426058 TI - Familial blood vessel tumors and subsequent cancers. AB - BACKGROUND: Population-based data on the familial risk for vascular tumors are largely lacking. Such data are important for clinical counseling and cancer genetics. METHODS: We used the Swedish Family-Cancer Database to calculate standardized incidence ratios for specific subtypes of vascular tumors in offspring using parents as probands. In addition, risks for second cancers were analyzed. RESULTS: Offspring hemangioblastoma in the nervous system was associated with parental kidney cancer and nervous system hemangioblastoma and hemangioma. Offspring nervous system hemangiopericytoma was associated with parental pituitary adenomas. Offspring angiosarcoma in the trunk and extremities was associated with maternal breast cancer. Second Kaposi's sarcoma, non Hodgkin's lymphoma, Hodgkin's disease and myeloma were increased following primary skin Kaposi's sarcoma. Kidney and endocrine gland tumors and nervous system hemangioblastomas and hemangiomas were in excess following primary nervous system hemangioblastoma and hemangioma. CONCLUSIONS: Our data showed that familial clustering of nervous system hemangioblastoma and hemangioma and the risks of subsequent cancers were primarily related to von-Hippel-Lindau disease. As a novel association, offspring nervous system hemangiopericytomas were in excess when parents were diagnosed with pituitary adenoma. Similarly, offspring angiosarcoma is associated with maternal breast cancer. Immunodeficiency may explain the excess of lymphoproliferative diseases after skin Kaposi's sarcoma. PMID- 17426059 TI - Gemcitabine, vinorelbine, and pegylated liposomal doxorubicin (GVD), a salvage regimen in relapsed Hodgkin's lymphoma: CALGB 59804. AB - BACKGROUND: Because of high single-agent activity and modest toxicity, we hypothesized the combination of gemcitabine (G), vinorelbine (V), and pegylated liposomal doxorubicin (D) would be an effective salvage therapy for Hodgkin's lymphoma (HL). PATIENTS AND METHODS: A total of 91 patients participated. GVD was administered on days 1 and 8 every 21 days at doses of G 1000 mg/m(2), V 20 mg/m(2), and D 15 mg/m(2) for transplant-naive patients, and G 800 mg/m(2), V 15 mg/m(2), and D 10 mg/m(2) for post-transplant patients. RESULTS: The dose limiting toxicity was mucositis for the transplant-naive patients and febrile neutropenia for post-transplant patients. The overall response rate (RR) for all patients was 70% [95% confidence interval (CI) 59.8, 79.7], with 19% complete remissions. The 4-year event-free and overall survival rates in transplant-naive patients treated with GVD followed by autologous transplant were 52% (95% CI 0.34, 0.68) and 70% (95% CI 0.49, 0.84), and in the patients in whom prior transplant failed, these were 10% (95% CI 0.03, 0.22) and 34% (95% CI 0.17, 0.52), respectively. CONCLUSIONS: GVD is a well-tolerated, active regimen for relapsed HL with results similar to those reported for more toxic regimens. High RRs in patients in whom prior transplant failed confirms this regimen's activity even in heavily pretreated patients. PMID- 17426060 TI - The role of Src in prostate cancer. AB - The Src family kinases (SFKs) are the largest family of nonreceptor protein tyrosine kinases and are responsible for signal transduction during many cellular activities, including differentiation, adhesion, and migration. Aberrant Src/SFK activity has been widely implicated in cancer development. Several lines of evidence indicate a role for SFKs in the development of prostate cancer, e.g. SFK overexpression in prostate cancer cell lines and tissues and reduced cancer cell proliferation, invasion, and migration following Src inhibition. In particular, Src may be involved in androgen-independent growth during advanced stages of disease. Src signaling is also a key pathway during normal and dysregulated bone functioning, and bone metastases are responsible for substantial morbidity in advanced prostate cancer. Src/SFK inhibition therefore represents a potentially useful therapeutic strategy for patients with various stages of prostate cancer. To date, four Src inhibitors have reached clinical trials. Of these, the broadest range of in vitro prostate cancer data are available for dasatinib, which inhibits several SFKs as well as other tyrosine kinases. Src inhibitors may be specifically evaluated in prostate cancer clinical trials in the near future. PMID- 17426061 TI - Vascular endothelial growth factor gene polymorphisms associated with prognosis for patients with gastric cancer. AB - BACKGROUND: The present study analyzed vascular endothelial growth factor (VEGF) gene polymorphisms and their impact on the prognosis for patients with gastric cancer. PATIENTS AND METHODS: Five hundred and three consecutive patients with surgically resected gastric adenocarcinoma were enrolled in the present study. The genomic DNA was extracted from paraffin-embedded tissue and four VEGF (-460T > C, -116G > A, +405G > C, and +936C > T) gene polymorphisms were determined using a polymerase chain reaction-restriction fragment length polymorphism assay. RESULTS: The survival analysis showed no association of three VEGF gene polymorphisms with the prognosis. For the +936C > T polymorphism, the T/T genotype, however, had a worse overall survival (OS) compared with the C/C genotype (P = 0.037). The -460 T/C or C/C genotype was a poor prognostic factor in patients with stage 0 or I gastric cancer (OS: hazard ratio (HR) = 3.96, disease-free survival (DFS): HR = 4.87). In the haplotype analysis, the CACC haplotype was associated with a significantly worse survival when compared with the TGGC haplotype (OS: HR = 1.72, DFS: HR = 1.73). CONCLUSIONS: VEGF gene polymorphisms were found to be an independent prognostic marker for patients with gastric cancer. Consequently, the analysis of VEGF gene polymorphisms can help identify patient subgroups at high risk of a poor disease outcome. PMID- 17426062 TI - Telemonitoring or structured telephone support programmes for patients with chronic heart failure: systematic review and meta-analysis. AB - OBJECTIVE: To determine whether remote monitoring (structured telephone support or telemonitoring) without regular clinic or home visits improves outcomes for patients with chronic heart failure. DATA SOURCES: 15 electronic databases, hand searches of previous studies, and contact with authors and experts. DATA EXTRACTION: Two investigators independently screened the results. REVIEW METHODS: Published randomised controlled trials comparing remote monitoring programmes with usual care in patients with chronic heart failure managed within the community. RESULTS: 14 randomised controlled trials (4264 patients) of remote monitoring met the inclusion criteria: four evaluated telemonitoring, nine evaluated structured telephone support, and one evaluated both. Remote monitoring programmes reduced the rates of admission to hospital for chronic heart failure by 21% (95% confidence interval 11% to 31%) and all cause mortality by 20% (8% to 31%); of the six trials evaluating health related quality of life three reported significant benefits with remote monitoring, and of the four studies examining healthcare costs with structured telephone support three reported reduced cost and one no effect. CONCLUSION: Programmes for chronic heart failure that include remote monitoring have a positive effect on clinical outcomes in community dwelling patients with chronic heart failure. PMID- 17426063 TI - Heavy metal--rely on gut feelings: novel diagnostic approach to test drug compliance in patients with lanthanum intake. PMID- 17426064 TI - Understanding the complexity of trans fatty acid reduction in the American diet: American Heart Association Trans Fat Conference 2006: report of the Trans Fat Conference Planning Group. AB - A 2-day forum was convened to discuss the current status and future implications of reducing trans fatty acids without increasing saturated fats in the food supply while maintaining functionality and consumer acceptance of packaged, processed, and prepared foods. Attendees represented the agriculture and oilseed industry and oil processing, food manufacturing, food service, government, food technology, and health and nutrition disciplines. Presentations included food science behind fatty acid technology, the health science of dietary fatty acids, alternatives to trans fatty acids, and the use of alternatives in food manufacturing and food service. The reduction of trans fatty acids in the food supply is a complex issue involving interdependent and interrelated stakeholders. Actions to reduce trans fatty acids need to carefully consider both intended and unintended consequences related to nutrition and public health. The unintended consequence of greatest concern is that fats and oils high in saturated fats, instead of the healthier unsaturated fats, might be used to replace fats and oils with trans fatty acids. Many different options of alternative oils and fats to replace trans fatty acids are available or in development. Decisions on the use of these alternatives need to consider availability, health effects, research and development investments, reformulated food quality and taste, supply-chain management, operational modifications, consumer acceptance, and cost. The conference demonstrated the value of collaboration between the food industry and health and nutrition professionals, and this conference model should be used to address other food development, processing, and/or technology issues. PMID- 17426065 TI - The efficacy of anti-TNF in rheumatoid arthritis, a comparison between randomised controlled trials and clinical practice. AB - BACKGROUND: Randomised controlled trials (RCTs) evaluating the efficacy of antagonists to tumour necrosis factor alpha (TNFalpha) showed high response percentages in the groups treated with active drugs. OBJECTIVE: To compare the efficacy of anti-TNF treatments for rheumatoid arthritis (RA) patients in RCTs and in daily clinical practice, with an emphasis on the efficacy for patients eligible and not eligible for RCTs of anti-TNF treatments. METHODS: First, randomised placebo-controlled trials written in English for etanercept, infliximab and adalimumab for patients with RA were selected by a systematic review. Second, the DREAM (Dutch Rheumatoid Arthritis Monitoring) register with patients starting for the first time on one of the TNF-blocking agents was used. Patient characteristics, doses of medication and co-medication as well as the ACR20 response percentages were compared between RCTs and DREAM data, stratified for trial eligibility. RESULTS: In 10 of 11 comparisons, the ACR20 response percentages were lower in daily clinical practice than in the RCT active drug group, which was significant in five of 11 comparisons. Only 34-79% of DREAM patients fulfilled the selection criteria for disease activity in the several RCTs examined. DREAM patients eligible for RCTs had higher response percentages than ineligible DREAM patients. ACR20 response percentages of eligible DREAM patients were comparable with the ACR20 response percentages of the RCT active drug group in 10 of 11 comparisons. CONCLUSION: The efficacy of TNF-blocking agents in RCTs exceeded the efficacy of these drugs in clinical practice. However, in clinical practice more patients with lower disease activity were treated with TNF-blocking agents compared with those treated in RCTs. For daily practice patients who were eligible for RCTs, responses were more similar to responses reached in RCTs. PMID- 17426066 TI - Mechano-sensitivity of epithelial sodium channels (ENaCs): laminar shear stress increases ion channel open probability. AB - Epithelial cells are exposed to a variety of mechanical forces, but little is known about the impact of these forces on epithelial ion channels. Here we show that mechanical activation of epithelial sodium channels (ENaCs), which are essential for electrolyte and water balance, occurs via an increased ion channel open probability. ENaC activity of heterologously expressed rat (rENaC) and Xenopus (xENaC) orthologs was measured by whole-cell as well as single-channel recordings. Laminar shear stress (LSS), producing shear forces in physiologically relevant ranges, was used to mechanically stimulate ENaCs and was able to activate ENaC currents in whole-cell recordings. Preceding pharmacological activation of rENaC with Zn2+ and xENaC with gadolinium and glibenclamide largely prevented LSS-activated currents. In contrast, proteolytic cleavage with trypsin potentiated the LSS effect on rENaC whereas the LSS effect on xENaC was reversed (inhibition of xENaC current). Further, we found that exposure of excised outside out patches to LSS led to an increased ion channel open probability without affecting the number of active channels. We suggest that mechano-sensitivity of ENaC may represent a ubiquitous feature for the physiology of epithelia, providing a putative mechanism for coupling transepithelial Na+ reabsorption to luminal transport. PMID- 17426067 TI - Membrane topology structure of human high-affinity, sodium-dependent dicarboxylate transporter. AB - High-affinity, sodium-dependent dicarboxylate transporter (NaDC3) is responsible for transport of Krebs cycle intermediates and may involve in regulation of aging and life span. Hydropathy analysis predicts that NaDC3 contains 11 or 12 hydrophobic transmembrane (TM) domains. However, the actual membrane topological structure of NaDC3 remains unknown. In this study, confocal immunofluorescence microscopy and membrane biotinylation of epitope-tagged N and C termini of NaDC3 provide evidence of an extracellular C terminus and an intracellular N terminus, indicating an odd number of transmembrane regions. The position of hydrophilic loops within NaDC3 was identified with antibodies against the loops domains combined with cysteine accessibility methods. A confocal image of membrane localization and transport activity assay of the cysteine insertion mutants show behavior similar to that of wild-type NaDC3 in transfected HEK293 cells, suggesting that these mutants retain a native protein configuration. We find that NaDC3 contains 11 transmembrane helices. The loops 1, 3, 5, 7, and 9 face the extracellular side, and loops 2, 4, 6, and 10 face the cytoplasmic side. A re entrant loop-like structure between TM8 and TM9 may protrude into the membrane. Our results support the topography of 11 transmembrane domains with an extracellular C terminus and an intracellular N terminus of NaDC3, and for the first time provide experimental evidence for a novel topological model for NaDC3. PMID- 17426068 TI - Rat liver sinusoidal endothelial cells survive without exogenous VEGF in 3D perfused co-cultures with hepatocytes. AB - Liver sinusoidal endothelial cells (SECs) are generally refractory to extended in vitro culture. In an attempt to recreate some features of the complex set of cues arising from the liver parenchyma, we cocultured adult rat liver SECs, identified by the expression of the marker SE-1, with primary adult rat hepatocytes in a 3D culture system that provides controlled microscale perfusion through the tissue mass. The culture was established in a medium containing serum and VEGF, and these factors were then removed to assess whether cells with the SE-1 phenotype could be supported by the local microenvironment in vitro. Rats expressing enhanced green fluorescent protein (EGFP) in all liver cells were used for isolation of the SE-1-positive cells added to cocultures. By the 13th day of culture, EGFP-expressing cells had largely disappeared from 2D control cultures but exhibited moderate proliferation in 3D perfused cultures. SE-1-positive cells were present in 3D cocultures after 13 days, and these cultures also contained Kupffer cells, stellate cells, and CD31-expressing endothelial cells. Global transcriptional profiling did not reveal profound changes between 2D and 3D cultures in expression of most canonical angiogenic factors but suggested changes in several pathways related to endothelial cell function. PMID- 17426069 TI - Low-dose remifentanil to suppress haemodynamic responses to noxious stimuli in cardiac surgery: a dose-finding study. AB - BACKGROUND: High-dose remifentanil (1-5 microg kg-1 min-1), commonly used for cardiac surgery, has been associated with muscle rigidity, hypotension, bradycardia, and reduced cardiac output. The aim of this study was to determine an optimal lower remifentanil dose, which should be accompanied by fewer adverse events, that still effectively suppresses haemodynamic responses to typical stressful stimuli (i.e. intubation, skin incision, and sternotomy). METHODS: Total i.v. anaesthesia consisted of a target-controlled propofol (2 microg ml-1) and a remifentanil infusion. Forty patients were allocated to receive either a constant infusion of remifentanil at 0.1 microg kg-1 min-1 or up-titrations to 0.2, 0.3, or 0.4 microg kg-1 min-1, respectively, 5 min before each stimulus. Subsequently, changes in heart rate and mean arterial blood pressure were recorded for 8 min. Increases exceeding 20% of baseline were considered to be of clinical relevance. Patients who exhibited these alterations were termed responders. RESULTS: The number of responders was less with the two higher remifentanil dosages (P<0.05) while propofol target doses could either be kept at the same level or even be reduced without affecting the plane of anaesthesia. Although single phenylephrine bolus had to be applied more frequently in these two groups (P<0.05), no severe haemodynamic depression was observed. CONCLUSIONS: Remifentanil at 0.3 and 0.4 microg kg-1 min-1 in combination with a target controlled propofol infusion in the pre-bypass period is well tolerated. It appears to mitigate potentially hazardous haemodynamic responses from stressful stimuli equally well as higher doses when compared with data from the literature. PMID- 17426070 TI - The rise and decline of tonsillectomy in twentieth-century America. AB - This article explores the rise and decline of tonsillectomy/adenoidectomy (T&A) in twentieth-century America. Between 1915 and the 1960s, T&A was the most frequently performed surgical procedure in the United States. Its rise was dependent on novel medical concepts, paradigms, and institutions that were in the process of reshaping the structure and practice of medicine. The driving force was the focal theory of infection, which assumed that circumscribed and confined infections could lead to systemic disease in any part of the body. The tonsils in particular were singled out as "portals of infection," and therefore their removal became a legitimate therapy. Nevertheless, what kinds of evidence could prove that tonsils were portals of infection? How could the effectiveness of tonsillectomy be determined? An inherent difficulty was the absence of any consensus on the criteria that would be employed to judge its efficacy. Yet tonsillectomy persisted despite ambiguous supportive evidence. Although criticisms of the procedure were common by the 1930s, its decline did not begin until well after 1945 and involved debates over the nature of evidence, the significance of clinical experience in the validation of a particular therapy, and the role of competing medical specialties. PMID- 17426071 TI - Prolonged renal failure secondary to antithymocyte globulin treatment in severe aplastic anemia. AB - OBJECTIVE: To report a case of acute renal failure in a patient with severe aplastic anemia after administration of antithymocyte globulin (ATG). CASE SUMMARY: A 41-year-old man diagnosed with severe aplastic anemia was treated with ATG and cyclosporine. After one dose of ATG (3012 mg, 40 mg/kg), the patient developed anuric acute renal failure, with serum creatinine 3.4 mg/dL (1.2 mg/dL at baseline) and blood urea nitrogen (BUN) 29 mg/dL (13 mg/dL at baseline), which required intermittent hemodialysis. Renal failure resolved with cessation of the drug, serum creatinine and BUN returned to baseline levels, and the patient no longer required hemodialysis. DISCUSSION: ATG is a purified and concentrated gamma globulin, primarily a monomeric immunoglobulin G from hyperimmune serum of horses. It is widely used to treat severe aplastic anemia and to manage acute transplant rejection. This patient had no other confounding factors for the cause of the renal failure. An objective causality assessment using the Naranjo probability scale suggested that ATG was the probable cause of the acute renal failure. Primary glomerular disease was not excluded, as a renal biopsy was not performed. CONCLUSIONS: The association between renal injury and administration of ATG remains unclear; therefore, we recommend that renal function be assessed and carefully monitored prior to and after administration of ATG. PMID- 17426072 TI - Effects of risperidone on lipid profile. PMID- 17426073 TI - Glycopeptide-induced neutropenia: cross-reactivity between vancomycin and teicoplanin. AB - OBJECTIVE: To report teicoplanin-related neutropenia that developed after an episode of neutropenia induced by vancomycin therapy. CASE SUMMARY: A 57-year-old female suffered from osteomyelitis of the left humerus, with a white blood cell (WBC) count of 2.8 x 10(3)/mm3 and absolute neutrophil count (ANC) of 0.28 x 10(3)/mm3, occurring after 24 days of vancomycin therapy. Vancomycin was changed to teicoplanin and the agranulocytosis resolved 4 days later. However, a new episode of neutropenia, with a WBC count of 2.8 x 10(3)/mm3 and ANC of 0.448 x 10(3)/mm3, occurred 11 days after teicoplanin initiation. Agranulocytosis resolved 4 days following withdrawal of teicoplanin. DISCUSSION: Because of the close time relationship between drug administration and the development of symptoms and signs, as well as between drug withdrawal and changes in WBC count and ANC, the episodes of neutropenia were suspected to be drug related. Teicoplanin-induced agranulocytosis that followed vancomycin-induced agranulocytosis suggests a possible cross-reactivity between the 2 drugs. Both reactions were categorized as probable according to the Naranjo probability scale. CONCLUSIONS: For all patients with vancomycin-induced neutropenia, possible cross-reactivity of teicoplanin should be monitored. PMID- 17426074 TI - Cefepime cerebrospinal fluid concentrations in neonatal bacterial meningitis. PMID- 17426075 TI - Effect of content and format of prescription drug labels on readability, understanding, and medication use: a systematic review. AB - OBJECTIVE: To evaluate the evidence regarding the optimal content and format of prescription labels that might improve readability, understanding, and medication use. DATA SOURCES: We performed a systematic review of randomized controlled trials, observational studies, and systematic reviews from MEDLINE and the Cochrane Database (1990-June 2005), supplemented by reference mining and reference lists from a technical expert panel. STUDY SELECTION: We selected studies that focused on the content of physician-patient communication about medications and the content and format of prescription drug labels. DATA EXTRACTION: Two reviewers extracted and synthesized information about study design, populations, and outcomes. DATA SYNTHESIS: Of 2009 articles screened, 36 that addressed the content of physician-patient communication about medications and 69 that were related to the content or format of medication labels met review criteria. Findings showed that patients request information about a drug's indication, expected benefits, duration of therapy, and a thorough list of potential adverse effects. The evidence about label format supports the use of larger fonts, lists, headers, and white space, using simple language and logical organization to improve readability and comprehension. Evidence was not sufficient to support the use of pictographic icons. Little evidence linked label design or content to measurable health outcomes, adherence, or safety. CONCLUSIONS: Evidence suggests that specific content and format of prescription drug labels facilitate communication with and comprehension by patients. Efforts to improve the labels should be guided by such evidence, although additional study assessing the influence of label design on medication-taking behavior and health outcomes is needed. Several policy options exist to require minimal standards to optimize medical therapy, particularly in light of the new Medicare prescription drug benefit. PMID- 17426076 TI - Drug-induced Sweet's syndrome. AB - OBJECTIVE: To systematically review the pertinent literature on drug-induced Sweet's syndrome (SS). DATA SOURCES: MEDLINE (1966-December 2006), International Pharmaceutical Abstracts (1970-December 2006), Science Citation Index (1945 December 2006), and EMBASE (1980-December 2006) were searched using the key terms Sweet's syndrome, drug-induced, and acute neutrophilic dermatitis. STUDY SELECTION AND DATA EXTRACTION: All case reports of drug-induced SS located using the above databases were collected for causality assessment. In addition, relevant articles regarding the various causes and presentations of SS were selected to provide background information. Bibliographies of all relevant articles were reviewed for additional citations. DATA SYNTHESIS: All case reports of drug-induced SS were evaluated against an expanded Naranjo scale with specific criteria for SS. Tables were developed listing key criteria for evaluating the case reports for causality. Data were evaluated by quantity and quality of evidence, and an assessment was made as to whether there was a feasible pharmacologic mechanism to explain causality. CONCLUSIONS: Granulocyte colony stimulating factor (G-CSF), all-trans retinoic acid (ATRA), and vaccines met 2 of 3 criteria for an association with SS. There are sufficient data and a plausible pharmacologic mechanism for G-CSF and ATRA. Vaccines meet the qualitative criteria and also have a plausible pharmacologic mechanism. The evidence regarding minocycline is of high quality; however, the quantity of evidence and a reasonable pharmacologic mechanism are lacking. A host of miscellaneous drugs have also been implicated in causing the disorder, all without sufficient evidence. PMID- 17426077 TI - Bazedoxifene: a third-generation selective estrogen receptor modulator for treatment of postmenopausal osteoporosis. AB - OBJECTIVE: To review clinical studies and other available literature regarding the development, pharmacology, toxicology, pharmacokinetics/pharmacodynamics, adverse effects, and place in therapy of bazedoxifene, a selective estrogen receptor modulator (SERM), currently in Phase III clinical trials for the treatment and prevention of postmenopausal osteoporosis. DATA SOURCES: A literature search was performed of PubMed (1966-February 2007), International Pharmaceutical Abstracts (1970-February 2007), Web of Science (1975-February 2007), Biological Abstracts (1926-2007), and Google Scholar (2001-February 2007) databases, using the search terms bazedoxifene, TSE-424, Indole-33, WAY-140424, selective estrogen receptor modulator, and SERM. In addition, product information was requested from the manufacturer, and www.clinicaltrials.gov was searched for unpublished Phase III clinical trials in progress. STUDY SELECTION AND DATA EXTRACTION: Articles on Phase I and II trials were selected for review, as well as articles discussing preclinical development of bazedoxifene. At the time of writing, no articles on Phase III trials were available for review. Abstracts of unpublished data were reviewed, as was information provided by the manufacturer. DATA SYNTHESIS: Bazedoxifene is a third-generation SERM currently in Phase III clinical trials. It has been found to act as an agonist on skeletal tissue, with bone turnover reduced by 20-25% with doses of 20 or 40 mg daily. In addition, bazedoxifene has been found to be an antagonist on breast tissue and uterine tissue, demonstrating inhibition of breast tissue proliferation and decreased endometrial stimulation as the dose is increased. CONCLUSIONS: Current literature suggests that bazedoxifene will likely be safe and effective when used in the treatment of postmenopausal osteoporosis. Completion of Phase III clinical trials will more fully elucidate the safety and efficacy profile of bazedoxifene, as well as more clearly define its place in therapy. PMID- 17426078 TI - Terbinafine-induced acute autoimmune hepatitis in the setting of hepatitis B virus infection. AB - OBJECTIVE: To report a case of terbinafine-induced autoimmune hepatitis in a patient with chronic hepatitis B infection. CASE SUMMARY: A 57-year-old Taiwanese male with chronic hepatitis B virus (HBV) began an oral regimen of terbinafine 250 mg once daily for dermatophyte toenail onychomycosis, despite the manufacturer's recommendation not to use the drug in patients with liver dysfunction. The patient's liver enzyme levels were within normal limits at initiation of therapy. Immediately prior to concluding the 12 week treatment course, he became anorexic with malaise and subsequently developed ascites and jaundice. After a visit to an outside emergency department and positively trending liver function test levels, he was referred to our liver clinic. The patient was taking no other medications or herbal supplements, did not drink alcohol, and did not appear to suffer a flare of HBV infection. The diagnosis was supported by the presence of transient autoantibodies and a liver biopsy consistent with acute autoimmune drug injury. Three weeks after terbinafine was discontinued, peak levels of aspartate aminotransferase (1282 IU/L), alanine aminotransferase (1044 IU/L), and bilirubin (5.9 mg/dL) were noted; his platelet level had decreased to 77 x 10(3)/mm3. He was treated with supportive care that included vitamin K for coagulopathy, diuretics for ascites, and adefovir to prevent hepatitis B exacerbation. The patient's liver function studies began to normalize 6 weeks after terbinafine was discontinued. DISCUSSION: Terbinafine induced hepatobiliary dysfunction, due to hepatocellular injury, cholestasis, or mixed form, has been reported, but this is the first case of autoimmune hepatitis supported by serologic, biochemical, and biopsy results. Use of the Naranjo probability scale revealed a probable relationship between the patient's hepatitis and terbinafine. Furthermore, the Roussel Uclaf Causality Assessment Method, a scoring system that specifically assesses the likelihood of drug induced elevated levels of liver-associated enzymes, also supported a probable relationship. The pathogenesis of most drug-induced autoimmune hepatitis remains speculative, likely involving hapten-carrier complex and the cytochrome P450 isoenzymes. In this patient, his chronic HBV carrier state may have predisposed him to this autoimmune reaction. CONCLUSIONS: Healthcare practitioners should heed the manufacturer's warning that terbinafine not be used in patients with underlying hepatic disease. PMID- 17426079 TI - Impact of left ventricular lead position in cardiac resynchronization therapy on left ventricular remodelling. A circumferential strain analysis based on 2D echocardiography. AB - AIMS: To assess if myocardial deformation imaging allows definition of an optimal left ventricular (LV) lead position with improved effectiveness of cardiac resynchronization therapy (CRT) on LV reverse remodelling. METHODS: Circumferential strain imaging based on tracking of acoustic markers within 2D echo images (GE Ultrasound) was performed in 47 heart failure patients (59 +/- 9 years, 28 men) at baseline, one day postoperatively, 3 and 10 months after initiation of CRT. Myocardial deformation imaging was used to determine(1) the segment with latest peak negative systolic circumferential strain prior to CRT, and(2) the segment with maximal temporal difference of peak strain before-to-on CRT as the segment with greatest benefit of CRT and assumed LV lead position. Anatomic LV lead position was determined by fluoroscopy. Optimal LV lead position was defined as concordance or immediate neighbouring of the segment with latest systolic strain prior to CRT and segment with assumed LV lead position. RESULTS: Agreement of assumed LV lead position based on strain analysis and LV lead position defined by fluoroscopy were high (kappa = 0.847). At 10 month follow-up, there was greater increase of EF (12 +/- 3 vs. 7 +/- 4%, P < 0.001), greater decrease of left ventricular end-diastolic volume (LVEDV) (23 +/- 8 vs. 13 +/- 7 mL, P < 0.001) and left ventricular end-systolic volume (LVESV) (42 +/- 10 vs. 27 +/- 8 mL, P < 0.001), and greater increase of VO(2)max (2.8 +/- 0.8 vs. 1.9 +/- 1.0 mL/kg/min, P = 0.035) in the optimal (n = 28 patients) compared to the non optimal LV lead position group (n = 19 patients). The distance between segment with latest systolic strain prior to CRT and segment with assumed LV lead position was the only independent predictor of DeltaLVEDV and DeltaLVESV at 10 month follow-up (R(2) = 0.2175, P = 0.0197) and (R(2) = 0.3774, P = 0.0054), respectively. CONCLUSION: Detailed analysis of the myocardial contraction sequence using circumferential strain imaging allows determination of the LV lead position in CRT. Optimal LV lead position in CRT defined by circumferential strain analysis results in greater improvement in LV function and more LV reverse remodelling than non-optimal LV lead position. PMID- 17426080 TI - Anticoagulants in heart disease: current status and perspectives. PMID- 17426081 TI - Germline NF1 mutational spectra and loss-of-heterozygosity analyses in patients with pheochromocytoma and neurofibromatosis type 1. AB - BACKGROUND: Neurofibromatosis type 1 (NF1) is a pheochromocytoma-associated syndrome. Because of the low prevalence of pheochromocytoma in NF1, we ascertained subjects by pheochromocytoma that also had NF1 in the hope of describing the germline NF1 mutational spectra of NF1-related pheochromocytoma. MATERIALS AND METHODS: An international registry for NF1-pheochromocytomas was established. Mutation scanning was performed using denaturing HPLC for intragenic variation and quantitative PCR for large deletions. Loss-of-heterozygosity analysis using markers in and around NF1 was performed. RESULTS: There were 37 eligible subjects (ages 14-70 yr). Of 21 patients with corresponding tumor available, 67% showed somatic loss of the nonmutated allele at the NF1 locus vs. 0 of 12 sporadic tumors (P = 0.0002). Overall, 86% of the 37 patients had exonic or splice site mutations, 14% large deletions or duplications; 79% of the mutations are novel. The cysteine-serine rich domain (CSR) was affected in 35% but the RAS GTPase activating protein domain (RGD) in only 13%. There did not appear to be an association between any clinical features, particularly pheochromocytoma presentation and severity, and NF1 mutation genotype. CONCLUSIONS: The germline NF1 mutational spectra comprise intragenic mutations and deletions in individuals with pheochromocytoma and NF1. NF1 mutations tended to cluster in the CSR over the RAS-GAP domain, suggesting that CSR plays a more prominent role in individuals with NF1-pheochromocytoma than in NF1 individuals without this tumor. Loss-of-heterozygosity of NF1 markers in NF1-related pheochromocytoma was significantly more frequent than in sporadic pheochromocytoma, providing further molecular evidence that pheochromocytoma is a true component of NF1. PMID- 17426082 TI - Maternal plasma concentrations of soluble endoglin in pregnancies with intrauterine growth restriction. AB - CONTEXT: Soluble endoglin (sEng), a coreceptor for TGF with antiangiogenic properties, acts synergistically with soluble fms-like tyrosine kinase 1 (sFlt1) to induce symptoms of HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome in animal models and to promote a preeclamptic phenotype. Pregnant women with preeclampsia show increased sEng concentrations in circulation, whereas the sEng increase is detectable months before the clinical onset of the disease. OBJECTIVE: The aim of the study was to determine whether maternal sEng is altered in pregnancies with normotensive intrauterine growth restriction (IUGR). DESIGN: sEng and sFlt1 were retrospectively determined by a commercial ELISA. PATIENTS: The study included 11 normotensive pregnancies with IUGR, 18 pregnancies with manifest preeclampsia, and 15 gestational-age-matched controls. RESULTS: Patients with preeclampsia showed significantly higher sEng concentrations compared with controls (57.0 ng/ml vs. 5.3 ng/ml; P < 0.001). Also IUGR pregnancies showed significantly elevated sEng concentrations (25.9 ng/ml; P < 0.001), but the levels were lower compared with the preeclamptic patients. There was a strong positive correlation between the sEng and sFlt1 concentration (Pearson 0.552; P < 0.01). Similar to sEng, the maternal sFlt1 concentration is highest in the preeclamptic patients (8388 vs. 2602 pg/ml; P < 0.01) but also significantly elevated in the IUGR patients (6952 pg/ml; P < 0.01). CONCLUSIONS: Pregnancy with IUGR, but without maternal symptoms, was characterized by elevated sEng concentrations in circulation. Although this finding is less pronounced when compared with preeclampsia, sEng seems to be involved in different clinical manifestations of placental pathology. PMID- 17426083 TI - Correlation between soluble endoglin, vascular endothelial growth factor receptor 1, and adipocytokines in preeclampsia. AB - CONTEXT: Recent reports have demonstrated that soluble endoglin (sEng), an antiangiogenic protein thought to impair TGF-beta binding to receptors, and soluble vascular endothelial growth factor receptor (sVEGFR)-1 play important roles in the pathophysiology of preeclampsia (PE). Moreover, insulin resistance, which is greatly influenced by adipocytokines, characterizes PE. OBJECTIVES: We examined possible links between sEng, VEGF, sVEGFR, and adipocytokines in the pathophysiology of PE. STUDY DESIGN: We performed a cross-sectional study in 30 PE patients and controls matched for gestational age and body mass index. Blood samples were collected soon after disease onset. We measured serum concentrations of leptin, adiponectin, sEng, VEGF, placental growth factor (PlGF), and sVEGFR [soluble fms-like tyrosine kinase 1 (sFlt-1) and soluble fetal liver kinase 1 (sFlk-1)], and examined the placental protein content of sEng and sFlt-1. RESULTS: sEng concentrations in PE patients (60.9 +/- 28.8 ng/ml) were significantly higher than those in controls (11.2 +/- 4.4 ng/ml). There was a significant correlation between sEng and sFlt-1 or PlGF. Moreover, there were significant differences in mean blood pressure between the high and low sEng groups, and in proteinuria between the high and low sFlt-1 groups, and significant differences in placental sEng and sFlt-1 contents between patients with and without severe hypertension or proteinuria. sEng was also correlated positively with adiponectin levels and negatively with the leptin to adiponectin ratio. CONCLUSIONS: Along with sFlt-1 and PlGF, sEng might play a role in the pathophysiology of PE, especially in elevating blood pressure, while the association with hypoadiponectinemia and the high leptin to adiponectin ratio in pregnancy seem to be risk factors for PE. PMID- 17426084 TI - High prevalence and mutual exclusivity of genetic alterations in the phosphatidylinositol-3-kinase/akt pathway in thyroid tumors. AB - CONTEXT: Genetic alterations in the phosphatidylinositol-3-kinase (PI3K)/Akt pathway and their role in thyroid tumor pathogenesis in Chinese people remain undefined. OBJECTIVE: The objective of the study was to examine the major genetic alterations and their relationship in the PI3K/Akt pathway in differentiated thyroid tumors in a Chinese cohort. DESIGN: We used real-time quantitative PCR for the analysis of PIK3CA copy gain and direct DNA sequencing for the detection of PIK3CA, RAS, and PTEN mutations on genomic DNA isolated from 234 thyroid tumors, including 31 follicular thyroid cancer (FTC), 141 papillary thyroid cancer (PTC), and 62 follicular thyroid adenoma (FTA). RESULTS: We found PIK3CA copy gain (defined as four or more copies) in nine of 31 FTC (29%), 20 of 141 PTC (14%), and five of 62 FTA (8%); PIK3CA gene mutations in four of 31 FTC (13%), one of 141 PTC (1%), and none of 62 FTA (0%); Ras mutations in three of 31 FTC (10%) and none of the 141 PTC and 62 FTA; and PTEN mutations in two of 31 FTC (6%) and none of 62 FTA (0%). Collectively, nine of 31 FTC (29%) vs. none of 62 FTA (0%) (P < 0.01) harbored one of the mutations, and when PIK3CA copy gain was included, 16 of 31 FTC (52%) vs. five of 62 FTA (8%) (P < 0.01) harbored any genetic alteration in the PI3K/Akt pathway. Mutual exclusivity was seen among all these PI3K/Akt pathway-related genetic alterations in all thyroid tumors except for two cases that harbored two genetic alterations. CONCLUSION: These data from a Chinese cohort provide further genetic evidence suggesting that dysregulated PI3K/Akt pathway plays a significant role in the pathogenesis of thyroid tumors, particularly FTC. PMID- 17426085 TI - Comparison of ethinyl-estradiol plus cyproterone acetate versus metformin effects on classic metabolic cardiovascular risk factors in women with the polycystic ovary syndrome. AB - CONTEXT: Oral contraceptives may worsen the metabolic profile of patients with polycystic ovary syndrome (PCOS), favoring the use of insulin sensitizers in these patients. OBJECTIVE: The aim of the study was to compare the effects of a contraceptive pill on metabolic classic cardiovascular risk factors with those of the insulin sensitizer metformin. DESIGN: We conducted a randomized, parallel, open-label clinical trial. SETTING: The study was conducted at an academic hospital. PATIENTS: Thirty-four consecutive PCOS patients were studied. INTERVENTIONS: Patients were randomized to oral treatment with metformin (850 mg twice daily) or with the Diane(35) Diario pill (35 microg of ethinyl-estradiol plus 2 mg of cyproterone acetate) for 24 wk. MAIN OUTCOME MEASURES: Hyperandrogenism, lipid profiles, and indexes of glucose tolerance and insulin sensitivity were measured at baseline and after 12 and 24 wk of treatment. RESULTS: Diane(35) Diario resulted in higher reductions in hirsutism score and serum androgen levels compared with metformin. Menstrual regularity was restored in all the patients treated with Diane(35) Diario compared with only 50% of those receiving metformin. Plasma apolipoprotein A-I and HDL-phospholipid levels increased with Diane(35) Diario, whereas metformin did not induce any change in the lipid profile. On the contrary, the insulin sensitivity index increased with metformin but did not change with Diane(35) Diario. No differences in the frequencies of abnormalities of glucose tolerance and dyslipidemia were found between both treatments. CONCLUSIONS: Diane(35) Diario appears to be superior to metformin for the control of hyperandrogenism and for the restoration of menstrual regularity in PCOS patients, and it is not associated with any clinically relevant worsening in the classic metabolic cardiovascular risk profile of these women. PMID- 17426086 TI - Effects of testosterone therapy on cardiovascular risk markers in androgen deficient women with hypopituitarism. AB - CONTEXT: Low-dose testosterone replacement therapy in women with relative androgen deficiency has been shown to have beneficial effects on body composition, bone mass, and psychosexual function. However, the safety of chronic testosterone administration on cardiovascular risk and insulin resistance is unknown. OBJECTIVE: The aim of the study was to determine the effects of physiological testosterone replacement on cardiovascular risk markers and insulin resistance in women. DESIGN: A 12-month, randomized, placebo-controlled study was conducted. SETTING: A General Clinical Research Center was the setting for the study. STUDY PARTICIPANTS: A total of 51 women of reproductive age with androgen deficiency due to hypopituitarism participated. INTERVENTION: Study participants were randomized to physiological testosterone administration, 300 mug daily, or placebo, by patch. MAIN OUTCOME MEASURES: We measured fasting glucose, fasting insulin, insulin-resistance homeostasis model of assessment (IRHOMA), quantitative insulin sensitivity check index (QUICKI), high-sensitivity C reactive protein, vascular cell adhesion molecule (VCAM), leptin, lipoprotein (a), apolipoprotein A1, and homocysteine. RESULTS: At 12 months, fasting insulin and IRHOMA were significantly lower in the testosterone compared with the placebo group, and there was a trend toward a higher QUICKI level at 12 months in the testosterone compared with the placebo group. These differences were no longer significant after controlling for baseline levels. We observed no effect, either positive or negative, of testosterone administration on high-sensitivity C reactive protein, VCAM leptin, lipoprotein (a), or apolipoprotein A1. CONCLUSIONS: Our data suggest that physiological testosterone replacement in women with hypopituitarism for 12 months does not increase, and may improve, insulin resistance. Chronic low-dose testosterone administration does not increase markers of cardiovascular disease reflecting several different mechanistic pathways. Large, randomized, placebo-controlled, long-term prospective studies are needed to determine whether low-dose testosterone replacement affects cardiovascular risk and event rates in women. PMID- 17426087 TI - The presence of the d3-growth hormone receptor polymorphism is negatively associated with fetal growth but positively associated with postnatal growth in healthy subjects. AB - CONTEXT: A common polymorphism in the GH receptor (GHR) gene has been linked to increased growth response in GH-treated patients. No former study has focused on the association to prenatal growth. OBJECTIVE: The aim of the study was to evaluate the association between the d3-GHR isoforms and spontaneous pre- and postnatal growth. DESIGN: A prospective study was conducted on third-trimester fetal growth velocity (FGV), birth weight, birth length, and postnatal growth. SETTING: The study was conducted at Copenhagen University Hospital. PARTICIPANTS: A total of 115 healthy adolescents were divided into those born small for gestational age (SGA) and appropriate for gestational age with or without intrauterine growth restriction. MAIN OUTCOME MEASURES: FGV was measured by serial ultrasonography, birth weight, birth length, and adolescent height. Isoforms of the d3-GHR gene (fl/fl, d3/fl, and d3/d3) were determined. RESULTS: The prevalence of the d3-GHR isoforms was 50% but differed among the groups (P = 0.006), with a high prevalence (88%) in the group born SGA with verified intrauterine growth restriction. The d3-GRH allele were associated with decreased third-trimester FGV (P = 0.05) in SGA subjects. In the entire cohort, carriers of the d3-GHR allele had a significantly increased height (-0.10 vs. 0.34 SD score; P = 0.017) and change in height from birth to adolescence compared with carriers of the full-length GHR allele (0.57 vs. -0.02 SD score; P = 0.005). CONCLUSIONS: This study showed an increased spontaneous postnatal growth velocity in the carriers of the d3-GHR allele. Interestingly, we found the opposite effect on prenatal growth in the SGA group, with a decreased FGV in carriers of the d3-GHR allele. PMID- 17426088 TI - Endocrine abnormalities in patients with Fanconi anemia. AB - BACKGROUND: Fanconi anemia (FA) is an inherited disorder with chromosomal instability, bone marrow failure, developmental defects, and a predisposition to cancer. Systematic and comprehensive endocrine function data in FA are limited. OBJECTIVE: We studied a cohort of FA patients enrolled in the National Cancer Institute's Inherited Bone Marrow Failure Syndrome study. STUDY DESIGN AND PATIENTS: Retrospective review of the medical records of 45 FA patients (ages 2 49 yr), 23 of whom were intensively evaluated at the National Institutes of Health. Anthropometric measurements, GH, IGF-I, IGF binding protein-3, thyroid, gonadal hormone, lipid levels, glucose homeostasis, brain imaging, and bone mineral density were obtained in these latter patients. RESULTS: Endocrine abnormalities were present in 73%, including short stature and/or GH deficiency (51%), hypothyroidism (37%), midline brain abnormalities (17%) (these patients had very short stature and 60% were GH-deficient); abnormal glucose/insulin metabolism (39%); obesity (27%); dyslipidemia (55%); and metabolic syndrome (21%). Patients with any endocrine abnormality were shorter than those without; only GH deficiency correlated significantly with short stature (P = 0.01). In addition, 65% of peripubertal or postpubertal patients had gonadal dysfunction. Ninety-two percent of the patients 18 yr or older had osteopenia or osteoporosis. CONCLUSIONS: Endocrine dysfunction is widespread in children and adults with FA; we expand the FA phenotype to include early onset osteopenia/osteoporosis and lipid abnormalities. Despite the reputation of FA as a progressive, lethal disease, proper management of the full spectrum of FA-related endocrinopathy offers major opportunities to reduce morbidity and improve quality of life. Our findings emphasize the need for comprehensive endocrine and metabolic evaluation and long-term follow-up in patients with FA. PMID- 17426089 TI - Cytotoxic T lymphocyte-associated molecule-4 gene polymorphism and hyperthyroid Graves' disease relapse after antithyroid drug withdrawal: a follow-up study. AB - CONTEXT: We previously showed an association between the exon1 +49 A/G single nucleotide polymorphism (SNP) and the relapse of Graves' disease (GD). The G allele was associated with early relapse. OBJECTIVE: In this follow-up study, we sought to replicate the result by genotyping nine additional polymorphisms and recruiting another 60 GD patients. DESIGN AND PARTICIPANTS: The GD patients were divided into three groups: recurred within 9 months, between 10-36 months, and more than 36 months. There were 65 patients with early recurrence, 55 with medium recurrence, and 88 with late recurrence. Although several SNPs were associated with recurrence, the most significant marker was still exon1 +49 A/G. Separate analysis of the genotypes for the 60 newly enrolled patients indicated that our present study was not biased by the previous samples. Once exon1 +49 A/G was included in the model to predict recurrence, other markers would not add more predictive information. Haplotype analysis did not show an additional value once exon1 +49 A/G was compulsorily included. RESULTS: Multivariate logistic regression analysis showed that GG genotype of exon1 +49 A/G SNP had an adjusted odds ratio of 2.2 (95% confidence interval, 1.1-4.4) compared with the combined group of GA plus AA. Other significant predictors were large goiter size at the end of the treatment and positive TSH-binding inhibitory Ig at the end of the treatment. CONCLUSIONS: This follow-up study confirms the usefulness of the exon1 +49 A/G SNP of the cytotoxic T lymphocyte-associated molecule-4 gene in predicting recurrence after cessation of treatment. There is no additional power by including other polymorphisms to predict recurrence. PMID- 17426090 TI - Effects of combined recombinant insulin-like growth factor (IGF)-I and IGF binding protein-3 in type 2 diabetic patients on glycemic control and distribution of IGF-I and IGF-II among serum binding protein complexes. AB - CONTEXT: Administration of recombinant human IGF-I (rhIGF-I)/recombinant human IGF binding protein-3 (rhIGFBP-3) to patients with type 2 diabetes improves blood glucose and enhances insulin sensitivity. The changes in various components of the IGF system that occur in response to rhIGF-I/rhIGFBP-3 as well as the minimum effective dose have not been determined. OBJECTIVES: The aim was to determine the dose of rhIGF-I/rh-IGFBP-3 necessary to achieve a significant decrease in glucose and to determine the changes that occur in the IGF-II and acid labile subunit in response to treatment. DESIGN: A total of 39 insulin-requiring type 2 diabetics were randomized to placebo or one of six groups that received different dosages of rhIGF-I/rhIGFBP-3. After 3 d in which insulin doses were adjusted to improve glucose control, a variable insulin dosage regimen was continued, and either placebo or one of six dosages (0.125-2.0 mg/kg.d) of rhIGF-I/rhIGFBP-3 was administered for 7 d. All subjects were hospitalized, and dietary intake as well as insulin dosage were controlled with instructions to treat to normal range targets. RESULTS: Fasting glucose was reduced in the groups that received either 1 (32 +/- 5% reduction) or 2 mg/kg.d (40 +/- 6% reduction) of the complex. Mean daily glucose (four determinations) was reduced by 26 +/- 4% in the 1 mg/kg group and by 33 +/- 5% in the 2 mg/kg group compared with 18 +/- 4% in the placebo group. Total serum IGF-I increased between 2.0 +/- 0.3- and 5.7 +/- 1.3-fold by d 8. IGFBP-3 concentrations increased significantly only in the 2 mg/kg group. IGF II concentrations declined to values that were between 27 +/- 4% and 64 +/- 7% below baseline. Acid labile subunit concentrations declined significantly in the three highest dose groups. The sum of the IGF-I + IGF-II concentrations was significantly increased at the two highest dosages. There were very few drug associated adverse events reported in this study with the exception of hypoglycemia, which occurred in 15 subjects who had received rhIGF-I/rhIGFBP-3 treatment. CONCLUSIONS: Administration of rhIGF-I/rhIGFBP-3 resulted in a redistribution of the amount of IGF-I and IGF-II that bound to IGFBP-3. Fasting and mean daily blood glucose were reduced significantly in the two highest dosage groups. The results suggest that both the total concentration of IGF-I as well as its distribution in blood may determine the extent to which insulin sensitivity is enhanced. PMID- 17426091 TI - Associations of adiponectin with body fat distribution and insulin sensitivity in nondiabetic Hispanics and African-Americans. AB - CONTEXT: Hypoadiponectinemia has emerged as an independent risk factor for type 2 diabetes and cardiovascular disease. Although associations of adiponectin with central obesity and insulin resistance have been reported, very little data are available from studies using detailed measures of insulin sensitivity (S(I)) and/or body fat distribution in ethnic groups at high risk for metabolic disease. OBJECTIVE: The aim of the study was to identify the correlates of adiponectin in 1636 nondiabetic Hispanics and African-Americans. DESIGN: A cross-sectional analysis of participants in the Insulin Resistance Atherosclerosis Family Study was conducted. S(I) was determined from frequently sampled iv glucose tolerance tests with minimal model analysis. Subcutaneous and visceral adipose tissues (SAT, VAT, respectively) were determined with computed tomography. Triglyceride, high-density lipoprotein, C-reactive protein, and adiponectin were measured in fasting samples. Generalized estimating equation (GEE) models were used to identify factors associated with adiponectin concentration. SETTING: A multicenter study using a family-based design was conducted. PARTICIPANTS: A total of 1636 nondiabetic Hispanic and African-American subjects participated. MAIN OUTCOME MEASURES: Circulating adiponectin concentration was measured. RESULTS: Age, female gender, high-density lipoprotein, SAT, and S(I) were positive independent correlates of adiponectin, whereas glucose, CRP, and VAT were negative independent correlates (all P < 0.05). Ethnicity was not an independent correlate of adiponectin in this model (P = 0.27); however, an ethnicity by VAT interaction term was retained, indicating a stronger negative association of VAT with adiponectin in African-Americans compared with Hispanics. CONCLUSION: Directly measured S(I), VAT, and SAT were independently correlated with adiponectin in Hispanic and African-American subjects. The inverse association of VAT with adiponectin was stronger in African-Americans compared with Hispanics, a finding that suggests possible ethnic differences in the effects of visceral obesity. PMID- 17426092 TI - Genes for enzymes regulating dehydroepiandrosterone sulfonation are associated with levels of dehydroepiandrosterone sulfate in polycystic ovary syndrome. AB - CONTEXT: The adrenal androgen (AA) metabolite dehydroepiandrosterone sulfate (DHEAS) is often elevated in women with polycystic ovary syndrome (PCOS); AA excess in PCOS appears to be, in part, a heritable trait. Dehydroepiandrosterone (DHEA) sulfonation is controlled by the enzymes DHEA sulfotransferase (SULT2A1) and steroid sulfatase (STS). Polymorphisms in these genes have not been evaluated as modulators of DHEAS level in PCOS. OBJECTIVE: The aim was to test the hypothesis that variants in the SULT2A1 and STS genes are associated with DHEAS levels in women with PCOS. DESIGN: Women with and without PCOS were genotyped for seven single nucleotide polymorphisms (SNPs) in SULT2A1 and seven SNPs in STS. SNPs and haplotypes were determined and tested for association with DHEAS. SETTING: Subjects were recruited from the reproductive endocrinology clinic at the University of Alabama at Birmingham; controls were recruited from the surrounding community. Genotyping took place at Cedars-Sinai Medical Center in Los Angeles. PARTICIPANTS: A total of 287 white women with PCOS and 187 controls participated in the study. MAIN MEASUREMENTS: SULT2A1 and STS genotype and DHEAS levels were measured. RESULTS: In women with PCOS, SNP rs182420 in SULT2A1 was associated with DHEAS (P = 0.0035). Two haplotypes carrying the minor allele of rs182420 were also associated with DHEAS (P = 0.04 each). Variants within STS were not associated with DHEAS level. No associations were observed in control women. CONCLUSION: This study presents genetic evidence suggesting a potential role of SULT2A1, but not STS, in the inherited AA excess of PCOS. PMID- 17426093 TI - Impact of fasting glycemia on short-term prognosis after acute myocardial infarction. AB - OBJECTIVE: The prognosis of patients with acute myocardial infarction (MI), according to the new criteria for impaired fasting glucose (IFG) (FG 100-126 mg/dl), has not been evaluated. RESEARCH DESIGN AND METHODS: A total of 2353 patients with acute MI and surviving at d 5 after admission were analyzed for short-term morbidity and mortality. FG was obtained at d 4 and 5. Patients were classified as diabetes mellitus (known diabetes or FG > or = 126 mg/dl), high IFG (110 < or = FG < 126 mg/dl), low IFG (100 < or = FG < 110 mg/dl), and normal fasting glucose (NFG) (FG < 100 mg/dl). RESULTS: Among the 2353 patients, 968 (41%) had diabetes mellitus, 262 (11%) had high IFG, 332 (14%) had low IFG, and 791 (34%) had NFG. Compared with NFG patients, 30-d cardiovascular mortality was increased in high but not low IFG subjects. In-hospital heart failure was increased in high IFG subjects (42 vs. 20% for NFG, P < 0.0001) but not low IFG subjects (21 vs. 20%). High IFG, but not low IFG, was an independent factor associated with 30-d cardiovascular mortality [odds ratio 2.33 (1.55-3.47)] and in-hospital heart failure [odds ratio 1.70 (1.36-2.07)]. The optimal threshold levels of FG on the receiver-operating characteristic curves were 114 and 112 mg/dl to predict mortality and in-hospital heart failure, respectively. CONCLUSION: The present study, based on a nonselected cohort of MI patients, underscores the high prevalence of IFG (25%) and highlights the clinical relevance of 110 mg/dl, but not 100 mg/dl, as a cutoff value to define the risk for worse outcome. PMID- 17426094 TI - Associations of serum thyrotropin concentrations with recurrence and death in differentiated thyroid cancer. AB - OBJECTIVE: The relation between serum TSH levels and risk for recurrence or thyroid carcinoma-related death in patients with differentiated thyroid carcinoma has only been studied to a limited extent. DESIGN: We conducted a single-center observational study in 366 consecutive patients with differentiated thyroid carcinoma, who had all been treated according to the same protocol for initial therapy and follow-up. Median duration of follow-up was 8.85 yr. METHODS: The relation between summarizing variables of unstimulated serum TSH concentrations (25th, 50th, and 75th percentiles, the percentage of suppressed and unsuppressed TSH values) and risk for recurrence or thyroid carcinoma-related death was analyzed by Cox survival analyses in patients with at least four TSH measurements. RESULTS: In Cox regression analysis, we found a positive association between serum TSH concentrations and risk for thyroid carcinoma related death and relapse, even in initially cured patients. The median of the individual TSH concentrations was the best indicator for thyroid carcinoma related death (hazard ratio 2.03; confidence interval 1.22-3.37) and relapse (hazard ratio 1.41; confidence interval 1.03-1.95). A threshold of 2 mU/liter differentiated best between relapse-free survival and thyroid carcinoma-related death or relapse. CONCLUSION: Our study supports current guidelines, which advise to aim at TSH levels in the low normal range in cured low-risk patients, whereas TSH levels should be suppressed in noncured or high-risk patients. PMID- 17426095 TI - Hypothalamic-pituitary-adrenal axis dysregulation and memory impairments in type 2 diabetes. AB - CONTEXT: There is evidence of both hypothalamic-pituitary-adrenocortical (HPA) axis and cognitive dysfunction in type 2 diabetes mellitus (T2DM). However, the exact nature and the associations between these abnormalities remain unclear. OBJECTIVES: The aim of the study was to characterize the nature of the HPA dysregulation in T2DM and ascertain whether impaired cognition in T2DM could be attributed to these abnormalities. DESIGN: A cross-sectional study was performed, contrasting matched groups on HPA axis function and cognition by using the combined dexamethasone (DEX)/CRH test and a neuropsychological battery assessing declarative and working memory, attention, and executive function. SETTING: The study was conducted in a research clinic in an academic medical center. PARTICIPANTS: Participants were volunteers functioning in the cognitively normal range. We studied 30 middle-aged individuals with T2DM, on average 7.5 yr since diabetes diagnosis, and 30 age-, gender-, and education-matched controls. MAIN OUTCOME MEASURES: Basal cortisol levels, cortisol levels during the DEX/CRH test, and performance on neuropsychological tests were measured. RESULTS: Individuals with T2DM had elevated basal plasma cortisol levels, higher levels after DEX suppression, and a larger response to CRH (all P A) were found in two different probands. Neither was found in the 74 nondiabetic controls and additional 270 healthy subjects of Thai origin. R164W segregated with diabetes in the family of the proband and in vitro studies showed that it impairs the repressor activity of PAX4 on the insulin and glucagon promoters. The remaining six variants were previously described and observed in both groups. One of them, R192H, was three times more frequent in MODY probands than in 342 nondiabetic controls (minor allele frequency = 0.196 vs. 0.064; P < 0.00001). The same variant was associated with a younger age at diagnosis among 254 Thai subjects with adult-onset type 2 diabetes (44.6 +/- 15 vs. 49.7 +/- 11 yr; P = 0.048). CONCLUSIONS: We have identified two possible pathogenic mutations of PAX4, R164W, and IVS7-1G>A. For one of these, we have shown evidence of segregation with diabetes and a functional impact on PAX4 activity. Single-nucleotide polymorphism R192H might influence the age at onset of diabetes. PMID- 17426100 TI - Visceral and subcutaneous adipose tissue assessed by magnetic resonance imaging in relation to circulating androgens, sex hormone-binding globulin, and luteinizing hormone in young men. AB - CONTEXT: No large studies of young men have examined circulating sex hormones in relation to visceral and sc adipose tissues. OBJECTIVE: The aim of this study was to investigate the role of visceral adipose tissue and sc adipose tissue on circulating sex hormones and the impact of obesity on sex hormone reference intervals. DESIGN, SETTING, AND PARTICIPANTS: Population-based study of 783 Danish 20- to 29-yr-old men was performed using dual-energy x-ray absorptiometry in all men and magnetic resonance imaging in 406 men. MAIN OUTCOME MEASURES: Total, bioavailable, and free testosterone, dihydrotestosterone (DHT), total and bioavailable estradiol, SHBG, and LH were measured. RESULTS: In multiple regressions, visceral adipose tissue was an independent, inverse correlate of bioavailable and free testosterone. Subcutaneous adipose tissue correlated negatively with SHBG and positively with bioavailable estradiol adjusted for total testosterone. Both visceral adipose tissue and sc adipose tissue correlated inversely with total testosterone and DHT. Adjusting for SHBG, only visceral adipose tissue remained significantly correlated. Low total testosterone in viscerally obese men was not accompanied by increased LH. The androgen reference intervals were significantly displaced toward lower limits in obese vs. nonobese men (total testosterone: 8.5-29.3 vs. 12.5-37.6 nmol/liter; bioavailable testosterone: 6.1-16.9 vs. 7.6-20.7 nmol/liter; free testosterone: 0.23-0.67 vs. 0.29-0.78 nmol/liter; and DHT: 0.63-2.5 vs. 0.85-3.2 nmol/liter), whereas total estradiol (36.5-166 pmol/liter) and bioavailable estradiol (23.4-120 pmol/liter) reference intervals were not. In obese men, 22.9% had total testosterone less than 12.5 nmol/liter. CONCLUSIONS: Visceral adipose tissues correlate independently with bioavailable and free testosterone in young men. The inverse relationship between total testosterone and sc adipose tissue seems to be accounted for by variations in SHBG. The reference intervals for total testosterone, bioavailable testosterone, free testosterone, and DHT are displaced toward lower limits in obese men. PMID- 17426101 TI - Relationships of plasma adiponectin level and adiponectin receptors 1 and 2 gene expression to insulin sensitivity and glucose and fat metabolism in monozygotic and dizygotic twins. AB - CONTEXT: Adiponectin is a key insulin-sensitizing adipokine acting on muscle metabolism via two specific receptors [adiponectin receptors 1 and 2 (AdipoR1 and AdipoR2, respectively)]. OBJECTIVES: The aim of the study was to investigate the genetic and nongenetic control of plasma adiponectin and muscle AdipoR1/R2 gene expression and the impact of these components on in vivo glucose and fat metabolism. DESIGN AND PARTICIPANTS: Plasma adiponectin and muscle gene expression of AdipoR1/R2 were measured before and during insulin infusion in 89 young and 69 elderly monozygotic and dizygotic twins. Insulin action, and glucose and fat oxidation rates were determined using hyperinsulinemic euglycemic clamps and indirect calorimetry. RESULTS: We demonstrated a genetic component in the control of plasma adiponectin and AdipoR1/R2 gene expression. Furthermore, levels of adiponectin and AdipoR1/R2 were influenced by age, sex, abdominal obesity, and aerobic capacity. Intrapair correlations in monozygotic twins indicated a nongenetic influence of birth weight on plasma adiponectin and AdipoR2 expression. Nonoxidative glucose metabolism was associated with AdipoR1 and plasma adiponectin, in young and elderly twins, respectively. In addition, plasma adiponectin was related to glucose and fat oxidation in younger subjects. CONCLUSIONS: Plasma adiponectin and muscle gene expression of its specific receptors are controlled by genetic and several specific nongenetic factors. The data suggest that the "adiponectin axis" plays a role in in vivo insulin action and nonoxidative glucose metabolism. PMID- 17426102 TI - Comparison of seven serum thyroglobulin assays in the follow-up of papillary and follicular thyroid cancer patients. AB - BACKGROUND: Serum thyroglobulin (Tg) is the marker of differentiated thyroid cancer after initial treatment and TSH stimulation increases its sensitivity for the diagnosis of recurrent disease. AIM: The goal of the study is to compare the diagnostic values of seven methods for serum Tg measurement for detecting recurrent disease both during L-T4 treatment and after TSH stimulation. METHODS: Thyroid cancer patients who had no evidence of persistent disease after initial treatment (total thyroidectomy and radioiodine ablation) were studied at 3 months on L-T4 treatment (Tg1) and then at 9-12 months after withdrawal or recombinant human TSH stimulation (Tg2). Sera with anti-Tg antibodies or with an abnormal recovery test result were excluded from Tg analysis with the corresponding assay. The results of serum Tg determination were compared to the clinical status of the patient at the end of follow-up. RESULTS: Thirty recurrences were detected among 944 patients. A control 131I total body scan had a low sensitivity, a low specificity, and a low clinical impact. Assuming a common cutoff for all Tg assays at 0.9 ng/ml, sensitivity ranged from 19-40% and 68-76% and specificity ranged from 92-97% and 81-91% for Tg 1 and Tg2, respectively. Using assays with a functional sensitivity at 0.2-0.3 ng/ml, sensitivity was 54-63% and specificity was 89% for Tg1. Using the two methods with a lowest functional sensitivity at 0.02 and 0.11 ng/ml resulted in a higher sensitivity for Tg1 (81% and 78%), but at the expense of a loss of specificity (42% and 63%); finally, for these two methods, using an optimized functional sensitivity according to receiver operating characteristic curves at 0.22 and 0.27 ng/ml resulted in a sensitivity at 65% and specificity at 85-87% for Tg1. CONCLUSION: Using an assay with a lower functional sensitivity may give an earlier indication of the presence of Tg in the serum on L-T4 treatment and may be used to study the trend in serum Tg without performing any TSH stimulation. Serum Tg determination obtained after TSH stimulation still permits a more reliable assessment of cure and patient's reassurance. PMID- 17426103 TI - Concurrent chemoradiotherapy for limited-disease small cell lung cancer in elderly patients aged 75 years or older. AB - BACKGROUND: The optimal treatment for limited-disease small cell lung cancer (LD SCLC) in patients aged 75 years or older remains unknown. METHODS: Elderly patients with LD-SCLC who were treated with chemoradiotherapy were retrospectively reviewed to evaluate their demographic characteristics and the treatment delivery, drug toxicities and antitumor efficacy. RESULTS: Of the 94 LD SCLC patients treated with chemotherapy and thoracic radiotherapy at the National Cancer Center Hospital between 1998 and 2003, seven (7.4%) were 75 years of age or older. All of the seven patients were in good general condition, with a performance status of 0 or 1. Five and two patients were treated with early and late concurrent chemoradiotherapy, respectively. While the four cycles of chemotherapy could be completed in only four patients, the full dose of radiotherapy was completed in all of the patients. Grade 4 neutropenia and thrombocytopenia were noted in seven and three patients, respectively. Granulocyte-colony stimulating factor support was used in five patients, red blood cell transfusion was administered in two patients and platelet transfusion was administered in one patient. Grade 3 or more severe esophagitis, pneumonitis and neutropenic fever developed in one, two and three patients, respectively, and one patient died of radiation pneumonitis. Complete response was achieved in six patients and partial response in one patient. The median survival time was 24.7 months, with three disease-free survivors for more than 5 years. CONCLUSION: Concurrent chemoradiotherapy promises to provide long-term benefit with acceptable toxicity for selected patients of LD-SCLC aged 75 years or older. PMID- 17426104 TI - Where are the patients in the quality of health care? PMID- 17426105 TI - Ketamine-induced neuronal cell death in the perinatal rhesus monkey. AB - Ketamine is widely used as a pediatric anesthetic. Studies in developing rodents have indicated that ketamine-induced anesthesia results in brain cell death. Additional studies are needed to determine if ketamine anesthesia results in brain cell death in the nonhuman primate and if so, to begin to define the stage of development and the duration of ketamine anesthesia necessary to produce brain cell death. Rhesus monkeys (N = 3 for each treatment and control group) at three stages of development (122 days of gestation and 5 and 35 postnatal days [PNDs]) were administered ketamine intravenously for 24 h to maintain a surgical anesthetic plane, followed by a 6-h withdrawal period. Similar studies were performed in PND 5 animals with 3 h of ketamine anesthesia. Animals were subsequently perfused and brain tissue processed for analyses. Ketamine (24-h infusion) produced a significant increase in the number of caspase 3-, Fluoro Jade C- and silver stain-positive cells in the cortex of gestational and PND 5 animals but not in PND 35 animals. Electron microscopy indicated typical nuclear condensation and fragmentation in some neuronal cells, and cell body swelling was observed in others indicating that ketamine-induced neuronal cell death is most likely both apoptotic and necrotic in nature. Ketamine increased N-methyl-D aspartate (NMDA) receptor NR1 subunit messenger RNA in the frontal cortex where enhanced cell death was apparent. Earlier developmental stages (122 days of gestation and 5 PNDs) appear more sensitive to ketamine-induced neuronal cell death than later in development (35 PNDs). However, a shorter duration of ketamine anesthesia (3 h) did not result in neuronal cell death in the 5-day-old monkey. PMID- 17426106 TI - Rodent carcinogenicity profile of the antidiabetic dual PPAR alpha and gamma agonist muraglitazar. AB - The carcinogenic potential of muraglitazar, a dual human peroxisome proliferator activated receptor alpha/gamma agonist, was evaluated in 2-year studies in mice (1, 5, 20, and 40 mg/kg) and rats (1, 5, 30, and 50 mg/kg). Benign gallbladder adenomas occurred at low incidences in male mice at 20 and 40 mg/kg (area under the curve [AUC] exposures > or = 62 times human exposure at 5 mg/day) and were considered drug related due to an increased incidence of gallbladder mucosal hyperplasia at these doses. There was a dose-related increased incidence of transitional cell papilloma and carcinoma of the urinary bladder in male rats at 5, 30, and 50 mg/kg (AUC exposures > or = 8 times human exposure at 5 mg/day). At 30 and 50 mg/kg, the urinary bladder tumors were accompanied by evidence of increased urine solids. Subsequent investigative studies established that the urinary bladder carcinogenic effect was mediated by urolithiasis rather than a direct pharmacologic effect on urothelium. Incidences of subcutaneous liposarcoma in male rats and subcutaneous lipoma in female rats were increased at 50 mg/kg (AUC exposures > or = 48 times human exposure at 5 mg/day) and attributed, in part, to persistent pharmacologic stimulation of preadipocytes. Toxicologically relevant nonneoplastic changes in target tissues included thinning of cortical bone in mice and hyperplastic and metaplastic adipocyte changes in mice and rats. Considering that muraglitazar is nongenotoxic, the observed tumorigenic effects in mice and rats have no established clinical relevance since they occurred at either clinically nonrelevant exposures (gallbladder and adipose tumors) or by a species-specific mechanism (urinary bladder tumors). PMID- 17426107 TI - Predicting age-appropriate pharmacokinetics of six volatile organic compounds in the rat utilizing physiologically based pharmacokinetic modeling. AB - The capability of physiologically based pharmacokinetic models to incorporate age appropriate physiological and chemical-specific parameters was utilized to predict changes in internal dosimetry for six volatile organic compounds (VOCs) across different ages of rats. Typical 6-h animal inhalation exposures to 50 and 500 ppm perchloroethylene, trichloroethylene, benzene, chloroform, methylene chloride, or methyl ethyl ketone (MEK) were simulated for postnatal day 10 (PND10), 2-month-old (adult), and 2-year-old (aged) rats. With the exception of MEK, predicted venous blood concentrations of VOCs in the aged rat were equal or up to 1.5-fold higher when compared to the adult rat at both exposure levels, whereas levels were predicted to be up to 3.8-fold higher in the case of PND10 at 50 ppm. Predicted blood levels of MEK were similar in the adult and aged rat, but were more than 5-fold and 30-fold greater for PND10 rats at 500 and 50 ppm, respectively, reflecting high water solubility along with lower metabolic capability and faster ventilation rate per unit body weight (BW) of PND10 animals. Steady-state blood levels of VOCs, simulated by modeling constant exposure, were predicted to be achieved in the order PND10 > adult > aged, largely due to increasing fat volume. The dose metric, total amount metabolized per unit liver volume was generally much lower in PND10 than in adult rats. The blood:air partition coefficient, fat volume, and fat blood flow were identified as critical determinants for the predicted differences in venous blood concentrations between the adult and aged. The lower metabolic capability, largely due to a smaller liver size, and faster ventilation rate per unit BW of PND10 animals contribute the most to the differences between PND10 and adult rats. This study highlights the pharmacokinetic differences and the relevant parameters that may contribute to differential susceptibility to the toxic effects of VOCs across life stages of the rat. PMID- 17426108 TI - Mode-of-action framework for evaluating the relevance of rodent forestomach tumors in cancer risk assessment. AB - Studies have shown that a majority of known human carcinogens also cause cancer in laboratory animals. The converse, however, is not as well established-known animal carcinogens are not equally predictive of human carcinogenicity. A particularly controversial aspect of interspecies extrapolation is application of rodent forestomach tumor data for predicting cancer risk in humans, given that a human counterpart for the rodent forestomach does not exist. Proliferative lesions in the rodent forestomach may result from a combination of factors related to route-specific tissue irritation and/or unnatural dosing regimens and are less likely to be relevant in evaluating human carcinogenic potential, particularly when tumors are exclusive to the forestomach. We review the comparative functional anatomy, physiology, tumor biology, tissue concordance, and historical regulatory practices in the use of forestomach tumors for cancer risk assessment, examining specific chemical examples. We also propose a standardized mode-of-action approach that combines multiple risk characterization criteria, including relevance to human exposure conditions, physiologically based toxicokinetics, genotoxicity, and comparative/mechanistic toxicology. Forestomach tumors associated with chronic irritation of the forestomach epithelium, particularly those induced by repeated oral gavage dosing, should not form the basis for carcinogenic classification or quantitative cancer potency estimates for humans. Genotoxic chemicals and those that cause tumors at multiple sites, at doses at or below the maximum tolerated dose, and in the absence of forestomach irritation, are more likely to be relevant human carcinogens. Cancer risk assessment that utilizes forestomach tumor data should consider relevant human exposures, systemic bioavailability, tissue dosimetry and concordance. PMID- 17426109 TI - High glucose induces IL-1beta expression in human monocytes: mechanistic insights. AB - Previously, IL-1beta secretion from Type 2 diabetic patients has been shown to be increased compared with controls. In this study, we aimed to delineate the mechanism of IL-1beta induction under high-glucose (HG) conditions in human monocytes. THP-1 cells cultured in normal glucose were treated with increasing concentrations of d-glucose (10-25 mM) for 6-72 h. IL-1beta and IL-1 receptor antagonist levels were measured by ELISA and Western blots, whereas mRNA was quantitated by RT-PCR. Specific inhibitors and small interfering RNAs of PKC, p38, ERK1/2, NF-kappaB, and NADPH oxidase were used to determine the mediators in parallel experiments under HG conditions. IL-1beta-secreted protein, cellular protein, and mRNA increase under HG conditions is time and dose dependent, with maximum increase at 15 mM (48 h; P < 0.05). IL-1 receptor antagonist release was time and dose dependent, similar to IL-1beta expression pattern; however, the molar ratio of IL-1beta to IL-1RA was increased. Data from inhibitor and small interfering RNA experiments indicate that IL-1beta release under HG is mediated by PKC-alpha, via phosphorylation of p38 MAPK, and ERK1/2 leading to NF-kappaB activation, resulting in increased mRNA and protein for IL-1beta. At the same time, it appears that NADPH oxidase via p47phox activates NF-kappaB, resulting in increased IL-1beta secretion. Data suggest that, under HG conditions, monocytes release significantly higher amounts of IL-1beta through multiple mechanisms, further compounding the disease progression. Targeting signaling pathways mediating IL-1beta release could result in the amelioration of inflammation and possibly diabetic vasculopathies. PMID- 17426110 TI - Sexually dimorphic responses to fat loss after caloric restriction or surgical lipectomy. AB - White adipose tissue is the principal site for lipid accumulation. Males and females maintain distinctive white adipose tissue distribution patterns. Specifically, males tend to accumulate relatively more visceral fat, whereas females accumulate relatively more subcutaneous fat. The phenomenon of maintaining typical sex-specific fat distributions suggests sex-specific mechanisms that regulate energy balance and adiposity. We used two distinct approaches to reduce fat mass, caloric restriction (CR), and surgical fat removal (termed lipectomy) and assessed parameters involved in the regulation of energy balance. We found that male and female mice responded differentially to CR- and to lipectomy-induced fat loss. Females decreased energy expenditure during CR or after lipectomy. In contrast, males responded by eating more food during food return after CR or after lipectomy. Female CR mice conserved subcutaneous fat, whereas male CR mice lost adiposity equally in the subcutaneous and visceral depots. In addition, female mice had a reduced capability to restore visceral fat after fat loss. After CR, plasma leptin levels decreased in male but not in female mice. The failure to increase food intake after returning to ad libitum intake in females could be due to the relatively stable levels of leptin. In summary, we have found sexual dimorphisms in the response to fat loss that point to important underlying differences in the strategies by which male and female mice regulate body weight. PMID- 17426111 TI - Physiological effects of nonthyroidal illness syndrome in patients after cardiac surgery. AB - In a prospective randomized placebo-controlled study, we assessed potential physiological effects of nonthyroidal illness syndrome (NTIS) in acute illness. Coronary artery bypass graft surgery was employed as a prospective model of acute illness and NTIS. Triiodothyronine (T(3)) or placebo was infused for 24 h after surgery, with a T(3) dose selected to maintain postoperative serum T(3) concentrations at preoperative levels. Patients were evaluated before coronary artery bypass graft and during the postoperative period. Cardiovascular function was monitored with Swan-Ganz catheter measurements and ECG. Urinary nitrogen excretion and L-[1-(13)C]leucine flux were used to evaluate protein metabolism. Serum measurements of relevant hormones, iron, and total iron-binding capacity were used to assess effects on sex steroid, growth hormone axis, and iron responses to illness. Cardiovascular function was not affected by T(3) infusion, except for a transient higher cardiac index in the T(3) group 6 h after surgery (3.04 +/- 0.12 for T(3) and 2.53 +/- 0.08 for placebo, P = 0.0016). Protein metabolism was not affected; changes in urinary nitrogen excretion and L-[1 (13)C]leucine flux were equivalent in the two groups (P = 0.35 and P = 0.95, respectively). No differences were observed in changes in testosterone, estrogens, growth hormone, insulin-like growth hormone I, iron, or total iron binding capacity between T(3) and placebo groups. We conclude that, in the early stages of major illness, the decrease in circulating T(3) concentrations in NTIS has only a minimal transient physiological impact on cardiac function and plays no significant role in protecting against protein catabolism or modulating other endocrine responses or iron responses to illness. PMID- 17426112 TI - Evaluation of intramyocellular lipid breakdown during exercise by biochemical assay, NMR spectroscopy, and Oil Red O staining. AB - The study compared the net decline of intramyocellular lipids (IMCL) during exercise (n = 18) measured by biochemical assay (BIO) and Oil Red O (ORO) staining on biopsy samples from vastus lateralis muscle and by (1)H-MR spectroscopy (MRS) sampled in an 11 x 11 x 18-mm(3) voxel in the same muscle. IMCL was measured before and after a 2-h cycling bout ( approximately 75% V(.)(O(2) peak)). ORO and MRS measurements showed substantial IMCL use during exercise of 31 +/- 12 and 47 +/- 6% of preexercise IMCL content. In contrast, use of BIO for IMCL determination did not reveal an exercise-induced breakdown of IMCL (2 +/- 9%, P = 0.29) in young healthy males. Correlations between different measures of exercise-induced IMCL degradation were low. Coefficients were 0.48 for MRS vs. ORO (P = 0.07) and were even lower for BIO vs. MRS (r = 0.38, P = 0.13) or ORO (r = 0.08, P = 0.78). This study demonstrates that different methods to measure IMCL in human muscles can result in different conclusions with regard to exercise-induced IMCL changes. MRS has the advantage that it is noninvasive, however, not fiber type specific and hampered by an at least 30-min delay in measurements after exercise completion and may overestimate IMCL use. BIO is the only quantitative method but is subject to variation when biopsies have different fiber type composition. However, BIO yields lower IMCL breakdown compared with ORO and MRS. ORO has the major advantage that it is fiber type specific, and it therefore provides information that is not available with the other methods. PMID- 17426113 TI - New functional aspects of the neuroendocrine marker secretagogin based on the characterization of its rat homolog. AB - Secretagogin is a recently cloned human beta-cell-expressed EF-hand Ca(2+) binding protein. Converging evidence indicates that it exerts Ca(2+) sensor activity and is involved in regulation of insulin synthesis and secretion. To obtain a potent tool for the extension of its functional analysis in rat in vitro systems, we cloned the rat homolog of human secretagogin. Using comparative sequence analysis, immunostaining, and immunoblotting, we demonstrated a high degree of sequence homology and similar tissue expression patterns of human and rat secretagogin. Highest rat secretagogin expression levels were found in pancreatic beta-cells. On the basis of newly generated anti-rat secretagogin antibodies, we established a rat secretagogin-specific sandwich capture ELISA and demonstrated release of secretagogin from viable Rin-5F cells. Dexamethasone treatment of Rin-5F cells resulted in an increased secretagogin release rate, which was inversely correlated with insulin secretion. In contrast, the secretagogin transcription rate was markedly reduced. This resulted in a decreased intracellular secretagogin content under the influence of dexamethasone. Sucrose gradient cell fractionation analysis of Rin-5F cells confirmed the predominant cytosolic localization of secretagogin, with only limited association of secretagogin with insulin granules. The loss of intracellular secretagogin after dexamethasone treatment affected predominantly the insulin granule-associated secretagogin fractions. The sequence homology and the comparable tissue expression patterns of human and rat secretagogin indicate conserved intracellular functions. The effects of dexamethasone on the total secretagogin content in Rin-5F cells and on its intracellular distribution might result in an impaired Ca(2+) sensitivity of dexamethasone-treated insulin secreting cells. PMID- 17426114 TI - Metabolic and histochemical characteristics of fat and muscle tissues in homozygous or heterozygous pigs for the body composition QTL located on chromosome 7. AB - Quantitative trait loci (QTL) influencing many traits including backfat thickness and carcass composition have been detected on porcine chromosome 7 (SSC7) in an F2 cross between Large White (LW) and Meishan (MS) pigs. However, the genes and controlled pathways underlying the QTL effects on body phenotype remain unknown. This study aimed at investigating the tissue characteristics at metabolic and cellular levels in pigs that were either homozygous or heterozygous for a body composition SSC7 QTL. A backcross pig (BC3) was first progeny tested to confirm its heterozygoty for the SSC7 QTL; results on all offspring (n = 80) confirmed the QTL effects on body fatness. This boar was then mated with three sows known to be heterozygous for this QTL. In the subset of pigs per genotype, we found that heterozygous LW(QTL7)/MS(QTL7) pigs had smaller adipocytes in backfat, together with a lower basal rate of glucose incorporation into lipids and lower activities of selected lipogenic enzymes in backfat isolated cells, compared with homozygous LW(QTL7)/LW(QTL7) pigs. A higher number of adipocytes was also estimated in backfat of LW(QTL7)/MS(QTL7) animals compared with LW(QTL7)/LW(QTL7) pigs. The SSC7 QTL did not influence oxidative and glycolytic metabolisms of longissimus and trapezius muscles, as estimated by the activities of specific energy metabolism enzymes, or the myofiber type properties. Altogether, this study provides new evidence for an altered adipocyte cellularity in backfat of pigs carrying at least one MS allele for the SSC7 QTL. Some candidate genes known for their functions on adipocyte growth and differentiation are suggested. PMID- 17426115 TI - Genome-wide analysis of PPARalpha activation in murine small intestine. AB - The peroxisome proliferator-activated receptor alpha (PPARalpha) is a fatty acid activated transcription factor that governs a variety of biological processes. Little is known about the role of PPARalpha in the small intestine. Since this organ is frequently exposed to high levels of PPARalpha ligands via the diet, we set out to characterize the function of PPARalpha in small intestine using functional genomics experiments and bioinformatics tools. PPARalpha was expressed at high levels in both human and murine small intestine. Detailed analyses showed that PPARalpha was expressed most highly in villus cells of proximal jejunum. Microarray analyses of total tissue samples revealed, that in addition to genes involved in fatty acid and triacylglycerol metabolism, transcription factors and enzymes connected to sterol and bile acid metabolism, including FXR and SREBP1, were specifically induced. In contrast, genes involved in cell cycle and differentiation, apoptosis, and host defense were repressed by PPARalpha activation. Additional analyses showed that intestinal PPARalpha-dependent gene regulation occurred in villus cells. Functional implications of array results were corroborated by morphometric data. The repression of genes involved in proliferation and apoptosis was accompanied by a 22% increase in villus height and a 34% increase in villus area of wild-type animals treated with WY14643. This is the first report providing a comprehensive overview of processes under control of PPARalpha in the small intestine. We show that PPARalpha is an important transcriptional regulator in small intestine, which may be of importance for the development of novel foods and therapies for obesity and inflammatory bowel diseases. PMID- 17426116 TI - Collateral density, remodeling, and VEGF-A expression differ widely between mouse strains. AB - Substantial variability exists in collateral density and ischemia-induced collateral growth among species. To begin to probe the underlying mechanisms, which are unknown, we characterized two mouse strains with marked differences in both parameters. Immediately after femoral artery ligation, collateral and foot perfusion were lower in BALB/c than C57BL/6 (P < 0.05 here and below), suggesting fewer pre-existing collaterals. This was confirmed with angiography and immunohistochemistry (approximately 35% fewer collaterals in the BALB/c's thigh). Recovery of hindlimb perfusion was attenuated in BALB/c, in association with 54% less collateral remodeling, reduced angiogenesis, greater ischemia, and more impaired hindlimb use. Densities of CD45+ and CD4+ leukocytes around collaterals increased similarly, but TNF-alpha expression was 50% lower in BALB/c, which may contribute to reduced collateral remodeling. In normal tissues, compared with C57BL/6, BALB/c exhibit an altered arterial branching pattern and, like skeletal muscle above, have 30% fewer collaterals in intestine and, remarkably, almost none in pial circulation, resulting in greatly impaired perfusion after cerebral artery occlusion. Ischemic induction of VEGF-A was attenuated in BALB/c. Analysis of a C57BL/6 x BALB/c recombinant inbred strain dataset identified a quantitative trait locus for VEGF-A mRNA abundance at or near the Vegfa locus that associates with lower expression in BALB/c. This suggests a cis-acting polymorphism in the Vegfa gene in BALB/c could contribute to reduced VEGF-A expression and, in turn, the above deficiencies in this strain. These findings suggest these strains offer a model to investigate genetic determinants of collateral formation and growth in ischemia. PMID- 17426117 TI - A promoting role of androgen receptor in androgen-sensitive and -insensitive prostate cancer cells. AB - Although the vital role of the androgen receptor (AR) has been well demonstrated in primary prostate cancers, its role in the androgen-insensitive prostate cancers still remains unclear. Here, we used a small hairpin RNA approach to directly assess AR activity in prostate cancer cells. Reduction of AR expression in the two androgen-sensitive prostate cancer cell lines, LNCaP and LAPC4, significantly decreased AR-mediated transcription and cell growth. Intriguingly, in two androgen-insensitive prostate cell lines, LNCaP-C42B4 and CWR22Rv1, knockdown of AR expression showed a more pronounced effect on AR-induced transcription and cell growth than androgen depletion. Using cDNA microarrays, we also compared the transcriptional profiles induced by either androgen depletion or AR knockdown. Although a significant number of transcripts appear to be regulated by both androgen depletion and AR knockdown, we observed a subset of transcripts affected only by androgen depletion but not by AR knockdown, and vice versa. Finally, we demonstrated a direct role for AR in promoting tumor formation and growth in a xenograft model. Taken together, our results elucidate an important role for the AR in androgen-insensitive prostate cancer cells, and suggest that AR can be used as a therapeutic target for androgen-insensitive prostate cancers. PMID- 17426118 TI - An exonuclease I hydrolysis assay for evaluating G-quadruplex stabilization by small molecules. AB - Telomere length homeostasis is a prerequisite for the generation and growth of cancer. In >85% cancer cells, telomere length is maintained by telomerase that add telomere repeats to the end of telomere DNA. Because the G-rich strand of telomere DNA can fold into G-quadruplex that inhibits telomerase activity, stabilizing telomere quadruplex by small molecules is emerging as a potential therapeutic strategy against cancer. In these applications, the specificity of small molecules toward quadruplex over other forms of DNA is an important property to ensure no processes other than telomere elongation are interrupted. The evaluating assays currently available more or less have difficulty identifying or distinguishing quadruplex-irrelevant effect from quadruplex stabilization. Here, we describe an exonuclease I hydrolysis assay that evaluates quadruplex stabilization by DNA-interacting compounds, discriminates inhibitory effect from different sources and helps determine the optimal compound concentration. PMID- 17426119 TI - Comparison of RNAi efficiency mediated by tetracycline-responsive H1 and U6 promoter variants in mammalian cell lines. AB - Conditional expression of short hairpin RNAs (shRNAs) to knock down target genes is a powerful tool to study gene function. The most common inducible expression systems are based on tetracycline-regulated RNA polymerase III promoters. During the last years, several tetracycline-inducible U6 and H1 promoter variants have been reported in different experimental settings showing variable efficiencies. In this study, we compare the most common variants of these promoters in several mammalian cell lines. For all cell lines tested, we find that several inducible U6 and H1 promoters containing single tetracycline operator (tetO) sequences show high-transcriptional background in the non-induced state. Promoter variants containing two tetO sequences show tight suppression of transcription in the non induced state, and high tet responsiveness and high gene knockdown efficiency upon induction in all cell lines tested. We report a variant of the H1 promoter containing two O2-type tetO sequences flanking the TATA box that shows little transcriptional background in the non-induced state and up to 90% target knockdown when the inducer molecule (dox-doxycycline) is added. This inducible system for RNAi-based gene silencing is a good candidate for use both in basic research on gene function and for potential therapeutic applications. PMID- 17426120 TI - Identification and characterization of OGG1 mutations in patients with Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) exhibit higher levels of 8-oxo-guanine (8 oxoG) DNA lesions in their brain, suggesting a reduced or defective 8-oxoG repair. To test this hypothesis, this study investigated 14 AD patients and 10 age-matched controls for mutations of the major 8-oxoG removal gene OGG1. Whereas no alterations were detected in any control samples, four AD patients exhibited mutations in OGG1, two carried a common single base (C796) deletion that alters the carboxyl terminal sequence of OGG1, and the other two had nucleotide alterations leading to single amino acid substitutions. In vitro biochemical assays revealed that the protein encoded by the C796-deleted OGG1 completely lost its 8-oxoG glycosylase activity, and that the two single residue-substituted OGG1 proteins showed a significant reduction in the glycosylase activity. These results were consistent with the fact that nuclear extracts derived from a limited number of AD patients with OGG1 mutations exhibited greatly reduced 8 oxoG glycosylase activity compared with age-matched controls and AD patients without OGG1 alterations. Our findings suggest that defects in OGG1 may be important in the pathogenesis of AD in a significant fraction of AD patients and provide new insight into the molecular basis for the disease. PMID- 17426121 TI - Flow cytometric analysis of DNA binding and cleavage by cell surface-displayed homing endonucleases. AB - LAGLIDADG homing endonucleases (LHEs) cleave 18-24 bp DNA sequences and are promising enzymes for applications requiring sequence-specific DNA cleavage amongst genome-sized DNA backgrounds. Here, we report a method for cell surface display of LHEs, which facilitates analysis of their DNA binding and cleavage properties by flow cytometry. Cells expressing surface LHEs can be stained with fluorescently conjugated double-stranded oligonucleotides (dsOligos) containing their respective target sequences. The signal is absolutely sequence specific and undetectable with dsOligos carrying single base-pair substitutions. LHE-dsOligo interactions facilitate rapid enrichment and viable recovery of rare LHE expressing cells by both fluorescence-activated cell sorting (FACS) and magnetic cell sorting (MACS). Additionally, dsOligos conjugated with unique fluorophores at opposite termini can be tethered to the cell surface and used to detect DNA cleavage. Recapitulation of DNA binding and cleavage by surface-displayed LHEs provides a high-throughput approach to library screening that should facilitate rapid identification and analysis of enzymes with novel sequence specificities. PMID- 17426122 TI - Selective use of multiple vitamin D response elements underlies the 1 alpha,25 dihydroxyvitamin D3-mediated negative regulation of the human CYP27B1 gene. AB - The human 25-hydroxyvitamin D3 (25(OH)D3) 1alpha-hydroxylase, which is encoded by the CYP27B1 gene, catalyzes the metabolic activation of the 25(OH)D3 into 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3), the most biologically potent vitamin D3 metabolite. The most important regulator of CYP27B1 gene activity is 1alpha,25(OH)2D3 itself, which down-regulates the gene. The down-regulation of the CYP27B1 gene has been proposed to involve a negative vitamin D response element (nVDRE) that is located approximately 500 bp upstream from transcription start site (TSS). In this study, we reveal the existence of two new VDR-binding regions in the distal promoter, 2.6 and 3.2 kb upstream from the TSS, that bind vitamin D receptor-retinoid X receptor complexes. Since the down regulation of the CYP27B1 gene is tissue- and cell-type selective, a comparative study was done for the new 1alpha,25(OH)2D3-responsive regions in HEK-293 human embryonic kidney and MCF-7 human breast cancer cells that reflect tissues that, respectively, are permissive and non-permissive to the phenomenon of 1alpha,25(OH)2D3-mediated down regulation of this gene. We found significant differences in the composition of protein complexes associated with these CYP27B1 promoter regions in the different cell lines, some of which reflect the capability of transcriptional repression of the CYP27B1 gene in these different cells. In addition, chromatin architecture differed with respect to chromatin looping in the two cell lines, as the new distal regions were differentially connected with the proximal promoter. This data explains, in part, why the human CYP27B1 gene is repressed in HEK-293 but not in MCF-7 cells. PMID- 17426123 TI - Stimulation of fission yeast and mouse Hop2-Mnd1 of the Dmc1 and Rad51 recombinases. AB - Genetic analysis of fission yeast suggests a role for the spHop2-Mnd1 proteins in the Rad51 and Dmc1-dependent meiotic recombination pathways. In order to gain biochemical insights into this process, we purified Schizosaccharomyces pombe Hop2-Mnd1 to homogeneity. spHop2 and spMnd1 interact by co-immunoprecipitation and two-hybrid analysis. Electron microscopy reveals that S. pombe Hop2-Mnd1 binds single-strand DNA ends of 3'-tailed DNA. Interestingly, spHop2-Mnd1 promotes the renaturation of complementary single-strand DNA and catalyses strand exchange reactions with short oligonucleotides. Importantly, we show that spHop2 Mnd1 stimulates spDmc1-dependent strand exchange and strand invasion. Ca(2+) alleviate the requirement for the order of addition of the proteins on DNA. We also demonstrate that while spHop2-Mnd1 affects spDmc1 specifically, mHop2 or mHop2-Mnd1 stimulates both the hRad51 and hDmc1 recombinases in strand exchange assays. Thus, our results suggest a crucial role for S. pombe and mouse Hop2-Mnd1 in homologous pairing and strand exchange and reveal evolutionary divergence in their specificity for the Dmc1 and Rad51 recombinases. PMID- 17426124 TI - A recombineering based approach for high-throughput conditional knockout targeting vector construction. AB - Functional analysis of mammalian genes in vivo is primarily achieved through analysing knockout mice. Now that the sequencing of several mammalian genomes has been completed, understanding functions of all the genes represents the next major challenge in the post-genome era. Generation of knockout mutant mice has currently been achieved by many research groups but only by making individual knockouts, one by one. New technological advances and the refinements of existing technologies are critical for genome-wide targeted mutagenesis in the mouse. We describe here new recombineering reagents and protocols that enable recombineering to be carried out in a 96-well format. Consequently, we are able to construct 96 conditional knockout targeting vectors simultaneously. Our new recombineering system makes it a reality to generate large numbers of precisely engineered DNA constructs for functional genomics studies. PMID- 17426125 TI - Novel DNA-binding element within the C-terminal extension of the nuclear receptor DNA-binding domain. AB - The heterodimer of the ecdysone receptor (EcR) and ultraspiracle (Usp), members of the nuclear receptors superfamily, is considered as the functional receptor for ecdysteroids initiating molting and metamorphosis in insects. Here we report the 1.95 A structure of the complex formed by the DNA-binding domains (DBDs) the EcR and the Usp, bound to the natural pseudopalindromic response element. Comparison of the structure with that obtained previously, using an idealized response element, shows how the EcRDBD, which has been previously reported to possess extraordinary flexibility, accommodates DNA-induced structural changes. Part of the C-terminal extension (CTE) of the EcRDBD folds into an alpha-helix whose location in the minor groove does not match any of the locations previously observed for nuclear receptors. Mutational analyses suggest that the alpha-helix is a component of EcR-box, a novel element indispensable for DNA-binding and located within the nuclear receptor CTE. This element seems to be a general feature of all known EcRs. PMID- 17426126 TI - An in vitro selection scheme for oligonucleotide probes to discriminate between closely related DNA sequences. AB - Using an in vitro selection, we have obtained oligonucleotide probes with high discriminatory power against multiple, similar nucleic acid sequences, which is often required in diagnostic applications for simultaneous testing of such sequences. We have tested this approach, referred to as iterative hybridizations, by selecting probes against six 22-nt-long sequence variants representing human papillomavirus, (HPV). We have obtained probes that efficiently discriminate between HPV types that differ by 3-7 nt. The probes were found effective to recognize HPV sequences of the type 6, 11, 16, 18 and a pair of type 31 and 33, either when immobilized on a solid support or in a reverse configuration, as well to discriminate HPV types from the clinical samples. This methodology can be extended to generate diagnostic kits that rely on nucleic acid hybridization between closely related sequences. In this approach, instead of adjusting hybridization conditions to the intended set of probe-target pairs, we 'adjust', through in vitro selection, the probes to the conditions we have chosen. Importantly, these conditions have to be 'relaxed', allowing the formation of a variety of not fully complementary complexes from which those that efficiently recognize and discriminate intended from non-intended targets can be readily selected. PMID- 17426127 TI - HIV controls the selective packaging of genomic, spliced viral and cellular RNAs into virions through different mechanisms. AB - In addition to genomic RNA, HIV-1 particles package cellular and spliced viral RNAs. In order to determine the encapsidation mechanisms of these RNAs, we determined the packaging efficiencies and specificities of genomic RNA, singly and fully spliced HIV mRNAs and different host RNAs species: 7SL RNA, U6 snRNA and GAPDH mRNA using RT-QPCR. Except GAPDH mRNA, all RNAs are selectively encapsidated. Singly spliced RNAs, harboring the Rev-responsible element, and fully spliced viral RNAs, which do not contain this motif, are enriched in virions to similar levels, even though they are exported from the nucleus by different routes. Deletions of key motifs (SL1 and/or SL3) of the packaging signal of genomic RNA indicate that HIV and host RNAs are encapsidated through independent mechanisms, while genomic and spliced viral RNA compete for the same trans-acting factor due to the presence of the 5' common exon containing the TAR, poly(A) and U5-PBS hairpins. Surprisingly, the RNA dimerization initiation site (DIS/SL1) appears to be the main packaging determinant of genomic RNA, but is not involved in packaging of spliced viral RNAs, suggesting a functional interaction with intronic sequences. Active and selective packaging of host and spliced viral RNAs provide new potential functions to these RNAs in the early stages of the virus life cycle. PMID- 17426128 TI - A hybrid CMV-H1 construct improves efficiency of PEI-delivered shRNA in the mouse brain. AB - RNA-interference-driven loss of function in specific tissues in vivo should permit analysis of gene function in temporally and spatially defined contexts. However, delivery of efficient short hairpin RNA (shRNA) to target tissues in vivo remains problematic. Here, we demonstrate that efficiency of polyethylenimine (PEI)-delivered shRNA depends on the regulatory sequences used, both in vivo and in vitro. When tested in vivo, silencing of a luciferase target gene by shRNA produced from a hybrid construct composed of the CMV enhancer/promoter placed immediately upstream of an H1 promoter (50%) exceeds that obtained with the H1 promoter alone (20%). In contrast, in NIH 3T3 cells, the H1 promoter was more efficient than the hybrid construct (75 versus 60% inhibition of target gene expression, respectively). To test CMV-H1 shRNA efficiency against an endogenous gene in vivo, we used shRNA against thyroid hormone receptor alpha1 (TRalpha1). When vectorized in the mouse brain, the hybrid construct strongly derepressed CyclinD1-luciferase reporter gene expression, CyclinD1 being a negatively regulated thyroid hormone target gene. We conclude that promoter choice affects shRNA efficiency distinctly in different in vitro and in vivo situations and that a hybrid CMV-H1 construct is optimal for shRNA delivery in the mouse brain. PMID- 17426129 TI - Biochemical and cellular characteristics of the 3' -> 5' exonuclease TREX2. AB - TREX2 is an autonomous nonprocessive 3' --> 5' exonuclease, suggesting that it maintains genome integrity. To investigate TREX2's biochemical and cellular properties, we show that endogenous TREX2 is expressed widely in mouse tissues and human cell lines. Unexpectedly, endogenous human TREX2 is predominantly expressed as a 30-kDa protein (not 26 kDa, as previously believed), which is likely encoded by longer isoforms (TREX2(L1) and/or TREX2(L2)) that possess similar capacity for self-association, DNA binding and catalytic activity. Site directed mutagenesis analysis shows that the three functional activities of TREX2 are distinct, yet integrated. Mutation of amino acids putatively important for homodimerization significantly impairs both DNA binding and exonuclease activity, while mutation of amino acids (except R163) in the DNA binding and exonuclease domains affects their corresponding activities. Interestingly, however, DNA binding domain mutations do not impact catalytic activity, while exonuclease domain mutations diminish DNA binding. To understand TREX2 cellular properties, we find endogenous TREX2 is down regulated during G2/M and nuclear TREX2 displays a punctate staining pattern. Furthermore, TREX2 knockdown reduces cell proliferation. Taken together, our results suggest that TREX2 plays an important function during DNA metabolism and cellular proliferation. PMID- 17426130 TI - Comparison of approaches for rational siRNA design leading to a new efficient and transparent method. AB - Current literature describes several methods for the design of efficient siRNAs with 19 perfectly matched base pairs and 2 nt overhangs. Using four independent databases totaling 3336 experimentally verified siRNAs, we compared how well several of these methods predict siRNA cleavage efficiency. According to receiver operating characteristics (ROC) and correlation analyses, the best programs were BioPredsi, ThermoComposition and DSIR. We also studied individual parameters that significantly and consistently correlated with siRNA efficacy in different databases. As a result of this work we developed a new method which utilizes linear regression fitting with local duplex stability, nucleotide position dependent preferences and total G/C content of siRNA duplexes as input parameters. The new method's discrimination ability of efficient and inefficient siRNAs is comparable with that of the best methods identified, but its parameters are more obviously related to the mechanisms of siRNA action in comparison with BioPredsi. This permits insight to the underlying physical features and relative importance of the parameters. The new method of predicting siRNA efficiency is faster than that of ThermoComposition because it does not employ time-consuming RNA secondary structure calculations and has much less parameters than DSIR. It is available as a web tool called 'siRNA scales'. PMID- 17426131 TI - DNA sequencing by MALDI-TOF MS using alkali cleavage of RNA/DNA chimeras. AB - Approaches developed for sequencing DNA with detection by mass spectrometry use strategies that deviate from the Sanger-type methods. Procedures demonstrated so far used the sequence specificity of RNA endonucleases, as unfortunately equivalent enzymes for DNA do not exist and therefore require transcription of DNA into RNA prior to fragmentation. We have developed a novel, rapid and accurate concept for DNA sequencing using mass spectrometry and RNA/DNA chimeras and applied it to sequence mitochondrial DNA. Our method is based on the preparation of a chimeric RNA/DNA with a DNA polymerase that also incorporates ribonucleotides. Sequencing is carried out with one ribonucleotide (ATP, CTP or GTP) and the other three nucleotides in their deoxyribo-form. The product is treated with alkali, which cleaves 3' of all ribonucleotides to form a terminal 3' phosphate. Conditions have been streamlined so that molecular, biological and alkali cleavage conditions are compatible with matrix-assisted laser desorption/ionization time-of-flight (MALDI) mass spectrometric analysis. Fragment analysis by MALDI MS provides a sequence-specific fingerprint, which allows the identification of differences between a reference and another sequence. Due to the mass profile, the position and kind of the mutation can be assigned. These differences between signatures are indicative of known, unidentified, rare and private mutations. This novel DNA sequencing protocol was applied to sequence the hypervariable region 1 (HV1) of mitochondrial DNA in 22 individuals. PMID- 17426132 TI - Nuclear localization of human DNA mismatch repair protein exonuclease 1 (hEXO1). AB - Human exonuclease 1 (hEXO1) is implicated in DNA mismatch repair (MMR) and mutations in hEXO1 may be associated with hereditary nonpolyposis colorectal cancer (HNPCC). Since the subcellular localization of MMR proteins is essential for proper MMR function, we characterized possible nuclear localization signals (NLSs) in hEXO1. Using fluorescent fusion proteins, we show that the sequence 418KRPR421, which exhibit strong homology to other monopartite NLS sequences, is responsible for correct nuclear localization of hEXO1. This NLS sequence is located in a region that is also required for hEXO1 interaction with hMLH1 and we show that defective nuclear localization of hEXO1 mutant proteins could be rescued by hMLH1 or hMSH2. Both hEXO1 and hMLH1 form complexes with the nuclear import factors importin beta/alpha1,3,7 whereas hMSH2 specifically recognizes importin beta/alpha3. Taken together, we infer that hEXO1, hMLH1 and hMSH2 form complexes and are imported to the nucleus together, and that redundant NLS import signals in the proteins may safeguard nuclear import and thereby MMR activity. PMID- 17426133 TI - The checkpoint clamp, Rad9-Rad1-Hus1 complex, preferentially stimulates the activity of apurinic/apyrimidinic endonuclease 1 and DNA polymerase beta in long patch base excision repair. AB - Growing evidence suggests that the Rad9-Rad1-Hus1 complex (the 9-1-1 complex), besides its functions in DNA damage sensing and signaling pathways, plays also a direct role in various DNA repair processes. Recent studies have demonstrated that the 9-1-1 complex physically and functionally interacts with several components of the base excision repair (BER) machinery namely DNA polymerase beta (Pol beta), flap endonuclease 1 (Fen 1), DNA ligase I (Lig I) and the MutY homologue of Schizosaccharomyces pombe. In this work, we found for the first time that the 9-1-1 complex interacts in vitro and in vivo with the apurinic/apyrimidinic endonuclease 1 (APE 1), an early component of BER, and can stimulate its AP-endonuclease activity. Moreover, we show that the 9-1-1 complex possesses a stimulatory effect on long patch base excision repair (LP-BER) reconstituted in vitro. The enhancement of LP-BER activity is due to the specific stimulation of the two early components of the repair machinery, namely APE 1 and Pol beta, suggesting a hierarchy of interactions between the 9-1-1 complex and the BER proteins acting in the repairosome. Overall, our results indicate that the 9-1-1 complex is directly involved in LP-BER, thus providing a possible link between DNA damage checkpoints and BER. PMID- 17426134 TI - The recombination-associated protein RdgC adopts a novel toroidal architecture for DNA binding. AB - RecA plays a central role in the nonmutagenic repair of stalled replication forks in bacteria. RdgC, a recombination-associated DNA-binding protein, is a potential negative regulator of RecA function. Here, we have determined the crystal structure of RdgC from Pseudomonas aeruginosa. The J-shaped monomer has a unique fold and can be divided into three structural domains: tip domain, center domain and base domain. Two such monomers dimerize to form a ring-shaped molecule of approximate 2-fold symmetry. Of the two inter-subunit interfaces within the dimer, one interface ('interface A') between tip/center domains is more nonpolar than the other ('interface B') between base domains. The structure allows us to propose that the RdgC dimer binds dsDNA through the central hole of approximately 30 A diameter. The proposed model is supported by our DNA-binding assays coupled with mutagenesis, which indicate that the conserved positively charged residues on the protein surface around the central hole play important roles in DNA binding. The novel ring-shaped architecture of the RdgC dimer has significant implications for its role in homologous recombination. PMID- 17426135 TI - Idefix insulator activity can be modulated by nearby regulatory elements. AB - Insulators play important roles in controlling gene activity and maintaining regulatory independence between neighbouring genes. In this article, we show that the enhancer-blocking activity of the insulator present within the LTR retrotransposon Idefix can be abolished if two copies of the region containing the insulator--specifically, the long terminal repeat (LTR)--are fused to the retrotransposon's 5' untranslated region (5' UTR). The presence of this combination of two [LTR-5' UTR] modules is a prerequisite for the loss of enhancer-blocking activity. We further show that the 5' UTR causes flanking genomic sequences to be displaced to the nuclear periphery, which is not observed when two insulators are present by themselves. This study thus provides a functional link between insulators and independent genomic modules, which may cooperate to allow the specific regulation of defined genomic loci via nuclear repositioning. It further illustrates the complexity of genomic regulation within a chromatic environment with multiple functional elements. PMID- 17426136 TI - Crystal structure of the third KH domain of human poly(C)-binding protein-2 in complex with a C-rich strand of human telomeric DNA at 1.6 A resolution. AB - KH (hnRNP K homology) domains, consisting of approximately 70 amino acid residues, are present in a variety of nucleic-acid-binding proteins. Among these are poly(C)-binding proteins (PCBPs), which are important regulators of mRNA stability and posttranscriptional regulation in general. All PCBPs contain three different KH domains and recognize poly(C)-sequences with high affinity and specificity. To reveal the molecular basis of poly(C)-sequence recognition, we have determined the crystal structure, at 1.6 A resolution, of PCBP2 KH3 domain in complex with a 7-nt DNA sequence (5'-AACCCTA-3') corresponding to one repeat of the C-rich strand of human telomeric DNA. The domain assumes a type-I KH fold in a betaalphaalphabetabetaalpha configuration. The protein-DNA interface could be studied in unprecedented detail and is made up of a series of direct and water mediated hydrogen bonds between the protein and the DNA, revealing an especially dense network involving several structural water molecules for the last 2 nt in the core recognition sequence. Unlike published KH domain structures, the protein crystallizes without protein-protein contacts, yielding new insights into the dimerization properties of different KH domains. A nucleotide platform, an interesting feature found in some RNA molecules, was identified, evidently for the first time in DNA. PMID- 17426137 TI - DNA structural deformations in the interaction of the controller protein C.AhdI with its operator sequence. AB - Controller proteins such as C.AhdI regulate the expression of bacterial restriction-modification genes, and ensure that methylation of the host DNA precedes restriction by delaying transcription of the endonuclease. The operator DNA sequence to which C.AhdI binds consists of two adjacent binding sites, O(L) and O(R). Binding of C.AhdI to O(L) and to O(L) + O(R) has been investigated by circular permutation DNA-bending assays and by circular dichroism (CD) spectroscopy. CD indicates considerable distortion to the DNA when bound by C.AhdI. Binding to one or two sites to form dimeric and tetrameric complexes increases the CD signal at 278 nm by 40 and 80% respectively, showing identical local distortion at both sites. In contrast, DNA-bending assays gave similar bend angles for both dimeric and tetrameric complexes (47 and 38 degrees, respectively). The relative orientation of C.AhdI dimers in the tetrameric complex and the structural role of the conserved Py-A-T sequences found at the centre of C-protein-binding sites are discussed. PMID- 17426138 TI - RNA helicase A interacts with divergent lymphotropic retroviruses and promotes translation of human T-cell leukemia virus type 1. AB - The 5' untranslated region (UTR) of retroviruses contain structured replication motifs that impose barriers to efficient ribosome scanning. Two RNA structural motifs that facilitate efficient translation initiation despite a complex 5' UTR are internal ribosome entry site (IRES) and 5' proximal post-transcriptional control element (PCE). Here, stringent RNA and protein analyses determined the 5' UTR of spleen necrosis virus (SNV), reticuloendotheliosis virus A (REV-A) and human T-cell leukemia virus type 1 (HTLV-1) exhibit PCE activity, but not IRES activity. Assessment of SNV translation initiation in the natural context of the provirus determined that SNV is reliant on a cap-dependent initiation mechanism. Experiments with siRNAs identified that REV-A and HTLV-1 PCE modulate post transcriptional gene expression through interaction with host RNA helicase A (RHA). Analysis of hybrid SNV/HTLV-1 proviruses determined SNV PCE facilitates Rex/Rex responsive element-independent Gag production and interaction with RHA is necessary. Ribosomal profile analyses determined that RHA is necessary for polysome association of HTLV-1 gag and provide direct evidence that RHA is necessary for efficient HTLV-1 replication. We conclude that PCE/RHA is an important translation regulatory axis of multiple lymphotropic retroviruses. We speculate divergent retroviruses have evolved a convergent RNA-protein interaction to modulate translation of their highly structured mRNA. PMID- 17426139 TI - Optimization and characterization of tRNA-shRNA expression constructs. AB - Expression of short hairpin RNAs via the use of PolIII-based transcription systems has proven to be an effective mechanism for triggering RNAi in mammalian cells. The most popular promoters for this purpose are the U6 and H1 promoters since they are easily manipulated for expression of shRNAs with defined start and stop signals. Multiplexing (the use of siRNAs against multiple targets) is one strategy that is being developed by a number of laboratories for the treatment of HIV infection since it increases the likelihood of suppressing the emergence of resistant virus in applications. In this context, the development of alternative small PolIII promoters other than U6 and H1 would be useful. We describe tRNA(Lys3)-shRNA chimeric expression cassettes which produce siRNAs with comparable efficacy and strand selectivity to U6-expressed shRNAs, and show that their activity is consistent with processing by endogenous 3' tRNAse. In addition, our observations suggest general guidelines for expressing effective tRNA-shRNAs with the potential for graded response, to minimize toxicities associated with competition for components of the endogenous RNAi pathway in cells. PMID- 17426140 TI - The TNF superfamily member LIGHT contributes to survival and activation of synovial fibroblasts in rheumatoid arthritis. AB - OBJECTIVES: The TNF superfamily member LIGHT has a T-cell co-stimulatory role and has previously been associated with inflammation and autoimmunity. To investigate its role in rheumatoid arthritis (RA), a disease where activated T cells contribute in a prominent way, we have analysed the expression of LIGHT and its receptors in RA and analysed its effects on synovial fibroblasts in vitro. METHODS: The expression of LIGHT was measured in synovial tissues and fluids and the receptors of LIGHT were detected on synovial fibroblasts derived from patients with RA and osteoarthritis (OA). The effects of recombinant LIGHT on the production of proinflammatory cytokines and proteases and on the apoptosis of synovial fibroblasts was assessed. RESULTS: LIGHT mRNA was present in synovial tissues of patients with RA but not with OA. Correspondingly, soluble LIGHT protein could be detected in RA synovial fluid samples at much higher levels than in synovial fluid from patients with OA. Immunohistochemical detection of LIGHT and analysis of synovial fluid cells by flow cytometry revealed CD4 T cells as the major source of LIGHT in the rheumatoid joint. Synovial fibroblasts from RA patients were found to express the LIGHT receptors HVEM and LTbetaR. Recombinant LIGHT induced RA synovial fibroblasts to upregulate MMP-9 mRNA, CD54 and IL-6 in an NF-kappaB-dependent fashion. In vitro, exposure of cultured synovial fibroblasts to LIGHT reduced FAS-mediated apoptosis significantly, without affecting the rate of spontaneous apoptosis. CONCLUSIONS: The results provide evidence for a novel T-cell-dependent activation of synovial fibroblasts by LIGHT in joints of patients with RA, contributing to an inflammatory and destructive phenotype. PMID- 17426141 TI - Using a self-reported functional score to assess disease progression in systemic sclerosis. AB - OBJECTIVES: This study compares the scleroderma Functional Score (FS) with the validated Disability Index of the Health Assessment Questionnaire (HAQ-DI) and other outcome measures. The aim is to determine if the FS is useful as an objective assessment tool for longitudinal evaluation of the functional impact of systemic sclerosis (SSc). METHODS: A cohort of 135 patients was studied (M:F, 15:120), with a mean age of 45.7 (S.D. = 13.2) at SSc disease onset. 69 (51%) had diffuse cutaneous scleroderma (dcSSc) and 66 (49%) had limited disease (lcSSc). The mean interval between the two assessments was 1.8 yrs (S.D. = 1.2). Functional impact was determined by evaluating archived self-reported questionnaires (FS, HAQ and scleroderma-VAS). Concurrent evaluation of the disease severity score was derived from clinical data stored in the hospital database and from medical case note reviews. RESULTS: At baseline, the mean FS was 11.0 (S.D. = 9.0) and at reassessment 12.0 (S.D. = 9.2). The mean absolute change in FS between the two assessments was 4.1 (S.D. = 4.9). With time 49% (n = 66) showed a clinically significant change in their functional ability with regard to FS, of these 29% (n = 39) worsened and 20% (n = 27) improved. There was an excellent cross-sectional correlation between the FS and the HAQ-DI (rho = 0.90; P < 0.0001). Also, a strong correlation between longitudinal change in these two outcome measures (rho = 0.59, P < 0.0001) was observed. CONCLUSIONS: This is the first longitudinal study of the scleroderma FS. It demonstrates that the FS can capture bidirectional and clinically significant changes in SSc related disability over time. The concurrent validity of the FS is asserted through its strong correlation with the HAQ-DI. The FS is a disease-specific, inexpensive and practical instrument for assessing functional status in SSc. It is a promising self-administered assessment tool for use in evaluating new SSc treatments. PMID- 17426142 TI - Differential P1 arginine and lysine recognition in the prototypical proprotein convertase Kex2. AB - The high-resolution crystal structure of kexin (Kex2) in complex with a peptidyl chloromethylketone inhibitor containing a noncognate lysine at the P(1) position provides the structural basis for the differential lysine/arginine selectivity that defines the prohormone (proprotein) convertase (PC) family. By comparison with the previous structures of Kex2 and furin, this structure of the acylated enzyme provides a basis for the observed decrease in the acylation rate with substrates containing a lysine at P(1) and the absence of an effect on the deacylation rate without involving mobility of the S(1) lid. The structure of the complex shows that a secondary subsite in the S(1) pocket is present, and that this site recognizes and binds the P(1) lysine in a more shallow fashion than arginine. This results in a displacement of the bound peptide away from the S385 nucleophile relative to substrates containing a P(1) arginine. It is concluded that this alternate binding site and resultant displacement of the scissile bond in the active site results in the observed decrease in the acylation rate. Studies of the inactivation kinetics of Kex2 by two peptidyl chloromethylketone inhibitors demonstrates that the selectivity between lysine and arginine at the P(1) position arises at the acylation step, consistent with what was observed with peptidyl substrates [Rockwell NC, Fuller RS (2001) J Biol Chem 276:38394 38399]. PMID- 17426143 TI - Replication fitness determines high virulence of influenza A virus in mice carrying functional Mx1 resistance gene. AB - The IFN-induced resistance factor Mx1 is a critical component of innate immunity against influenza A viruses (FLUAV) in mice. Animals carrying a wild-type Mx1 gene (Mx1(+/+)) differ from regular laboratory mice (Mx1(-/-)) in that they are highly resistant to infection with standard FLUAV strains. We identified an extraordinary variant of the FLUAV strain, A/PR/8/34 (H1N1) (designated hvPR8), which is unusually virulent in Mx1(+/+) mice. hvPR8 was well controlled in Mx1(+/+) but not Mx1(-/-) mice provided that the animals were treated with IFN before infection, indicating that hvPR8 exhibits normal sensitivity to growth restriction by Mx1. hvPR8 multiplied much faster than standard PR8 early in infection because of highly efficient viral gene expression in infected cells. Studies with reassortant viruses containing defined genome segments of both hvPR8 and standard PR8 demonstrated that the HA, neuraminidase, and polymerase genes of hvPR8 all contributed to virulence, indicating that efficient host cell entry and early gene expression renders hvPR8 highly pathogenic. These results reveal a surprisingly simple concept of how influenza viruses may gain virulence and illustrate that high speed of virus growth can outcompete the antiviral response of the infected host. PMID- 17426144 TI - Vesicle formation from the nuclear membrane is induced by coexpression of two conserved herpesvirus proteins. AB - Although the nuclear envelope is a dynamic structure that disassembles and reforms during mitosis, the formation of membranous vesicles derived from the nuclear envelope has not yet been described in noninfected cells. However, during herpesvirus maturation, intranuclear capsids initiate transit to the cytosol for final maturation by budding at the inner nuclear membrane. Two conserved herpesvirus proteins are required for this primary envelopment, designated in the alphaherpesviruses as pUL31 and pUL34. Here, we show that simultaneous expression of pUL31 and pUL34 of the alphaherpesvirus pseudorabies virus in stably transfected rabbit kidney cells resulted in the formation of vesicles in the perinuclear space that resemble primary envelopes without a nucleocapsid. They contain pUL31 and pUL34 as shown by immunolabeling and are derived from the nuclear envelope. Thus, coexpression of only two conserved herpesvirus proteins without any other viral factor is sufficient to induce the formation of vesicles from the nuclear membrane. This argues for the contribution of cellular factors in this process either recruited from their natural cytoplasmic location or not yet identified as components of the nuclear compartment. PMID- 17426145 TI - Lack of Spem1 causes aberrant cytoplasm removal, sperm deformation, and male infertility. AB - We identified a previously uncharacterized gene, spermatid maturation 1 (Spem1), encoding a protein exclusively expressed in the cytoplasm of steps 14-16 elongated spermatids in the mouse testis. This protein contains no known functional domains and is highly conserved across mammalian species. Male mice deficient in Spem1 were completely infertile because of deformed sperm characterized by a bent head wrapped around by the neck and the middle piece of the tail. We show that lack of Spem1 causes failure of the cytoplasm to become loose and detach from the head and the neck region of the developing spermatozoa. Retained cytoplasmic components mechanically obstruct the straightening of the sperm head and the stretching of the growing tail, leading to the bending of the head in the neck, followed by the wrapping of the head by the neck or the middle piece of the sperm tail. Our study reveals that proper cytoplasm removal is a genetically regulated process requiring the participation of Spem1 and that lack of Spem1 causes sperm deformation and male infertility. PMID- 17426147 TI - Microfossil evidence for pre-Columbian maize dispersals in the neotropics from San Andres, Tabasco, Mexico. AB - The history of maize (Zea mays L.) is one of the most debated topics in New World archaeology. Molecular and genetic studies indicate that maize domestication took place in tropical southwest Mexico. Although archaeological evidence for the evolution of maize from its wild ancestor teosinte has yet to be found in that poorly studied region, other research combining paleoecology and archaeology is documenting the nature and timing of maize domestication and dispersals. Here we report a phytolith analysis of sediments from San Andres, Tabasco, that confirms the spread of maize cultivation to the tropical Mexican Gulf Coast >7,000 years ago ( approximately 7,300 calendar years before present). We review the different methods used in sampling, identifying, and dating fossil maize remains and compare their strengths and weaknesses. Finally, we examine how San Andres amplifies the present evidence for widespread maize dispersals into Central and South America. Multiple data sets from many sites indicate that maize was brought under cultivation and domesticated and had spread rapidly out of its domestication cradle in tropical southwest Mexico by the eighth millennium before the present. PMID- 17426148 TI - Beta-arrestin is a necessary component of Wnt/beta-catenin signaling in vitro and in vivo. AB - The Wnt/beta-catenin signaling pathway is crucial for proper embryonic development and tissue homeostasis. The phosphoprotein dishevelled (Dvl) is an integral part of Wnt signaling and has recently been shown to interact with the multifunctional scaffolding protein beta-arrestin. Using Dvl deletion constructs, we found that beta-arrestin binds a region N-terminal of the PDZ domain of Dvl, which contains casein kinase 1 (CK1) phosphorylation sites. Inhibition of Wnt signaling by CK1 inhibitors reduced the binding of beta-arrestin to Dvl. Moreover, mouse embryonic fibroblasts lacking beta-arrestins were able to phosphorylate LRP6 in response to Wnt-3a but decreased the activation of Dvl and blocked beta-catenin signaling. In addition, we found that beta-arrestin can bind axin and forms a trimeric complex with axin and Dvl. Furthermore, treatment of Xenopus laevis embryos with beta-arrestin morpholinos reduced the activation of endogenous beta-catenin, decreased the expression of the beta-catenin target gene, Xnr3, and blocked axis duplication induced by X-Wnt-8, CK1epsilon, or DshDeltaDEP, but not by beta-catenin. Thus, our results identify beta-arrestin as a necessary component for Wnt/beta-catenin signaling, linking Dvl and axin, and open a vast array of signaling avenues and possibilities for cross-talk with other beta-arrestin-dependent signaling pathways. PMID- 17426146 TI - Genomic resources for songbird research and their use in characterizing gene expression during brain development. AB - Vocal learning and neuronal replacement have been studied extensively in songbirds, but until recently, few molecular and genomic tools for songbird research existed. Here we describe new molecular/genomic resources developed in our laboratory. We made cDNA libraries from zebra finch (Taeniopygia guttata) brains at different developmental stages. A total of 11,000 cDNA clones from these libraries, representing 5,866 unique gene transcripts, were randomly picked and sequenced from the 3' ends. A web-based database was established for clone tracking, sequence analysis, and functional annotations. Our cDNA libraries were not normalized. Sequencing ESTs without normalization produced many developmental stage-specific sequences, yielding insights into patterns of gene expression at different stages of brain development. In particular, the cDNA library made from brains at posthatching day 30-50, corresponding to the period of rapid song system development and song learning, has the most diverse and richest set of genes expressed. We also identified five microRNAs whose sequences are highly conserved between zebra finch and other species. We printed cDNA microarrays and profiled gene expression in the high vocal center of both adult male zebra finches and canaries (Serinus canaria). Genes differentially expressed in the high vocal center were identified from the microarray hybridization results. Selected genes were validated by in situ hybridization. Networks among the regulated genes were also identified. These resources provide songbird biologists with tools for genome annotation, comparative genomics, and microarray gene expression analysis. PMID- 17426149 TI - Absence of dopamine D2 receptors unmasks an inhibitory control over the brain circuitries activated by cocaine. AB - Cocaine is a psychostimulant and a drug widely abused by humans. Cocaine elicits its effects primarily by blocking the activity of the dopamine (DA) transporter, leading to elevated levels of extracellular DA in areas receiving dopaminergic innervation, with the consequent activation of DA receptors. Cocaine, however, also elevates other neurotransmitter levels, leading to a general activation of interconnected brain circuitries. Studies aimed at unraveling the molecular mechanisms underlying the effects of cocaine have shown a leading role of DA D1 receptors in the cascade of cellular events elicited by this drug. In this study, we have analyzed the acute response to cocaine in animals deleted for the expression of DA D2 receptors (D2R), an inhibitor of DA signaling. Importantly, we show that although D1 receptor-mediated functions are preserved and even enhanced in D2R-/- mutants, the behavioral response to acute cocaine administration is severely altered. In addition, c-fos response to acute cocaine administration, in contrast to wild-type mice, is absent in D2R-/- mutants. Our findings show that the absence of D2R, very likely through a presynaptic mechanism, uncovers an inhibitory signaling pathway normally masked by the activity of this receptor on brain circuitries engaged by abused drugs. PMID- 17426150 TI - Blocking S-adenosylmethionine synthesis in yeast allows selenomethionine incorporation and multiwavelength anomalous dispersion phasing. AB - Saccharomyces cerevisiae is an ideal host from which to obtain high levels of posttranslationally modified eukaryotic proteins for x-ray crystallography. However, extensive replacement of methionine by selenomethionine for anomalous dispersion phasing has proven intractable in yeast. We report a general method to incorporate selenomethionine into proteins expressed in yeast based on manipulation of the appropriate metabolic pathways. sam1(-) sam2(-) mutants, in which the conversion of methionine to S-adenosylmethionine is blocked, exhibit reduced selenomethionine toxicity compared with wild-type yeast, increased production of protein during growth in selenomethionine, and efficient replacement of methionine by selenomethionine, based on quantitative mass spectrometry and x-ray crystallography. The structure of yeast tryptophanyl-tRNA synthetase was solved to 1.8 A by using multiwavelength anomalous dispersion phasing with protein that was expressed and purified from the sam1(-) sam2(-) strain grown in selenomethionine. Six of eight selenium residues were located in the structure. PMID- 17426151 TI - Seismological support for the metastable superplume model, sharp features, and phase changes within the lower mantle. AB - Recently, a metastable thermal-chemical convection model was proposed to explain the African Superplume. Its bulk tabular shape remains relatively stable while its interior undergoes significant stirring with low-velocity conduits along its edges and down-welling near the middle. Here, we perform a mapping of chemistry and temperature into P and S velocity variations and replace a seismically derived structure with this hybrid model. Synthetic seismogram sections generated for this 2D model are then compared directly with corresponding seismic observations of P (P, P(C)P, and PKP) and S (S, S(C)S, and SKS) phases. These results explain the anticorrelation between the bulk velocity and shear velocity and the sharpness and level of SKS travel time delays. In addition, we present evidence for the existence of a D" triplication (a putative phase change) beneath the down-welling structure. PMID- 17426152 TI - Gorilla-like anatomy on Australopithecus afarensis mandibles suggests Au. afarensis link to robust australopiths. AB - Mandibular ramus morphology on a recently discovered specimen of Australopithecus afarensis closely matches that of gorillas. This finding was unexpected given that chimpanzees are the closest living relatives of humans. Because modern humans, chimpanzees, orangutans, and many other primates share a ramal morphology that differs from that of gorillas, the gorilla anatomy must represent a unique condition, and its appearance in fossil hominins must represent an independently derived morphology. This particular morphology appears also in Australopithecus robustus. The presence of the morphology in both the latter and Au. afarensis and its absence in modern humans cast doubt on the role of Au. afarensis as a modern human ancestor. The ramal anatomy of the earlier Ardipithecus ramidus is virtually that of a chimpanzee, corroborating the proposed phylogenetic scenario. PMID- 17426153 TI - Aldolase provides an unusual binding site for thrombospondin-related anonymous protein in the invasion machinery of the malaria parasite. AB - An actomyosin motor located underneath the plasma membrane drives motility and host-cell invasion of apicomplexan parasites such as Plasmodium falciparum and Plasmodium vivax, the causative agents of malaria. Aldolase connects the motor actin filaments to transmembrane adhesive proteins of the thrombospondin-related anonymous protein (TRAP) family and transduces the motor force across the parasite surface. The TRAP-aldolase interaction is a distinctive and critical trait of host hepatocyte invasion by Plasmodium sporozoites, with a likely similar interaction crucial for erythrocyte invasion by merozoites. Here, we describe 2.4-A and 2.7-A structures of P. falciparum aldolase (PfAldo) obtained from crystals grown in the presence of the C-terminal hexapeptide of TRAP from Plasmodium berghei. The indole ring of the critical penultimate Trp-residue of TRAP fits snugly into a newly formed hydrophobic pocket, which is exclusively delimited by hydrophilic residues: two arginines, one glutamate, and one glutamine. Comparison with the unliganded PfAldo structure shows that the two arginines adopt new side-chain rotamers, whereas a 25-residue subdomain, forming a helix-loop-helix unit, shifts upon binding the TRAP-tail. The structural data are in agreement with decreased TRAP binding after mutagenesis of PfAldo residues in and near the induced TRAP-binding pocket. Remarkably, the TRAP- and actin binding sites of PfAldo seem to overlap, suggesting that both the plasticity of the aldolase active-site region and the multimeric nature of the enzyme are crucial for its intriguing nonenzymatic function in the invasion machinery of the malaria parasite. PMID- 17426154 TI - Spatial orientation of mitochondrial processing peptidase and a preprotein revealed by fluorescence resonance energy transfer. AB - Mitochondrial processing peptidase (MPP), which is composed of heterodimeric alpha-MPP and beta-MPP subunits. It specifically recognizes mitochondrial preproteins and removes their basic N-terminal signal prepeptides. In order to elucidate the spatial orientation of the preproteins toward MPP, which has been missed by crystal structures of a yeast MPP including a synthetic prepeptide in its acidic proteolytic chamber, we analysed the fluorescence resonance energy transfer (FRET) between EGFP fused to a yeast aconitase presequence (preEGFP) and regiospecific 7-dietylamino-3-(4'-maleimidyl phenyl)-4-methyl coumarin (CPM) labelled yeast MPPs. FRET efficiencies of 65 and 55% were observed between the EGFP chromophore and CPM-Ser(84) and -Lys(156) of beta-MPP, respectively, leading to calculated distances between the molecules of 48 and 50 A, respectively. Considering the FRET results and the structural validity based on the crystal structure of the MPP-presequence complex, a plausible model of preEGFP associated with MPP was constructed in silico. The modelled structure indicated that amino acid residues on the C-terminal side of the cleavage site in the preprotein were orientated tail out from the large cavity of MPP and interacted with the glycine rich loop of alpha-MPP. Thus, MPP orientates preproteins at the specific cleft between the catalytic domain and the flexible glycine-rich loop which seems to pinch the extended polypeptide. PMID- 17426155 TI - Double-labelled in situ hybridization reveals the lack of co-localization of mRNAs for the circadian neuropeptide PDF and FMRFamide in brains of the flies Musca domestica and Drosophila melanogaster. AB - Many lines of evidence have suggested that neuropeptides other than pigment dispersing factor (PDF) are involved in regulating insect circadian rhythms, and FMRFamide-related peptides are additional candidates acting as such neuromodulators. Double-immunolabelling in insect brains with anti-crustacean beta-PDH and anti-FMRFamide antibodies had previously suggested that insect PDF and FMRFamide-like peptides may coexist in the same cells. However, it is critical for this kind of comparative investigations to use antibodies of proven specificity, to eliminate the possibility of both reciprocal cross-reactivity and the detection of unknown peptides. In the present study, we achieved the cDNA cloning of an fmrf mRNA from the housefly Musca domestica, for which co localization of FMRFamide and PDF peptides was previously suggested. In order to examine the possible co-expression of this gene with the pdf gene, we carried out double-labelled in situ hybridization for simultaneous detection of both pdf and fmrf mRNAs in housefly, Musca brains. The results clearly indicated that they occur in distinctly different cells. This was also proven for the fruit fly Drosophila melanogaster by similar double-labelled in situ hybridization. The results thus revealed no reason to evoke the physiological release of FMRFamide and PDF peptides from the same neurons. PMID- 17426156 TI - The unheralded and essential contribution of the field worker in tropical medicine: a committed technician making a difference in Corte de Pedra, Brazil. PMID- 17426157 TI - Sulfadoxine-pyrimethamine efficacy and selection of Plasmodium falciparum DHFR mutations in Burkina Faso before its introduction as intermittent preventive treatment for pregnant women. AB - Sulfadoxine-pyrimethamine efficacy was determined with a 28-day follow-up in 97 children between 6 months and 15 years of age. The polymerase chain reaction (PCR)-corrected treatment failure was 8.2% and the uncorrected was 21.6%. The presence of the dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) mutations linked to sulfadoxine-pyrimethamine resistance before and after treatment was determined by PCR-restriction fragment length polymorphism (RFLP) and by a fluorogenic PCR assay. Before treatment, the prevalence of the triple DHFR mutations was higher among the patients having had a recurrent parasitemia (either recrudescence or new infection; 28.6% versus 9.3%), although the difference was not significant (P = 0.1). The double mutation Ala-436/Gly-437 was observed in 67% of samples, whereas no Glu-540 mutation was found. After treatment, the triple DHFR mutation was found in 76.2% of patients with recurrent parasitemia, recrudescence, and new infection alike. Such high prevalence of mutant parasites indicates that sulfadoxine-pyrimethamine should not be used as monotherapy. PMID- 17426158 TI - Unusual pattern of Plasmodium falciparum drug resistance in the northwestern Peruvian Amazon region. AB - High levels of Plasmodium falciparum resistance to both chloroquine (CQ) and sulfadoxine-pyrimethamine (SP) have been documented throughout the Amazon Basin of South America. Because of reports about the persistent efficacy of both of these drugs in the northwestern Peruvian Amazon region, we carried out an evaluation of the therapeutic efficacy of chloroquine (25 mg/kg) and SP (25 mg/kg of the sulfadoxine component) for the treatment of uncomplicated P. falciparum infections at two sites: Ullpayacu and Pampa Hermoza/Alianza. A total of 111 patients were enrolled. Only 5 (14.3%) of the 35 patients who received CQ had an adequate clinical and parasitologic response (ACPR). Six subjects (17%) had early treatment failure, 1 (2.9%) had late clinical failure, and 23 (65.7%) had late parasitologic failure (LPF). Of the subjects treated with SP, 92.3% had ACPR and 7.7% had LPF. Based on these findings, it is clear that there are at least limited areas within the Peruvian Amazon region where P. falciparum strains continue to be sensitive to SP. PMID- 17426159 TI - Malaria risk factors in Amerindian children in French Guiana. AB - Malaria is a major public health problem in French Guiana. This study was conducted in children <1-5 years of age in Camopi, an Amerindian village in eastern French Guiana. Medical, environmental, and behavioral predictive factors of malaria were studied using the Kaplan-Meier method and Cox modeling. Variables included were clearing vegetation around the home (hazard ratio [HR] = 0.62, 95% confidence interval [CI] = 0.43-0.88 for 50-75% cleared and HR = 0.5, 95% CI = 0.31-0.81 for > 75% cleared) relative to homes surrounded by vegetation; distance of a home from a river (HR = 0.56, 95% CI = 0.37-0.85 for distances between 20 and 40 meters, HR = 0.72, 95% CI = 0.47-1.09 for distances between 40 and 80 meters, HR = 0.52, 95% CI = 0.28-0.94 for distances between 80 and 120 meters, and HR = 0.5, 95% CI = 0.30-0.86 for distances > 120 meters) relative to distances < 20 meters; the number of occupants in the home (HR = 1.54, 95% CI = 0.98-2.44 for 7 occupants, HR = 1.9, 95% CI = 1.29-2.81 for 8-11 occupants, and HR = 2.03, 95% CI = 1.27-3.23 for > 11 occupants); clothing (HR = 0.64, 95% CI = 0.46-0.90 for children wearing western-style clothes relative to those wearing the traditional kalimbe), and ethnicity (Wayampi children had a lower hazard of malaria relative to Emerillon children: HR = 0.55, 95% CI = 0.40-0.80). The environment near the home was strongly associated with malaria. This suggests that simple pragmatic protection measures would be useful in Camopi. PMID- 17426160 TI - Age-dependent distribution of Plasmodium falciparum gametocytes quantified by Pfs25 real-time QT-NASBA in a cross-sectional study in Burkina Faso. AB - Sexual stages of Plasmodium falciparum play a key role in the transmission of malaria. Studies on gametocytes are generally based on microscopic detection, but more sensitive detection methods for P. falciparum gametocytes frequently detect sub-patent gametocytes. We used Pfs25 mRNA quantitative-nucleic acid sequence based amplification (QT-NASBA) to quantify gametocytes in 412 samples from a cross-sectional study in Burkina Faso, covering all age groups, to determine age related patterns in gametocyte carriage and gametocyte density. The more sensitive QT-NASBA technique gave estimates of gametocyte prevalence 3.3-fold higher than microscopy (70.1% versus 21.4%, respectively). Prevalence of gametocytes significantly decreased with age. Our data suggest that asexual parasite densities are primarily responsible for the age-related decrease of gametocyte prevalence, possibly because of developing asexual stage immunity. Gametocyte densities decrease also with age, primarily because of decreasing asexual parasite densities; only a small but significant age effect on gametocyte density may be caused by developing sexual stage-specific immunity. PMID- 17426161 TI - Southeast Asian ovalocytosis and pregnancy in a malaria-endemic region of Papua New Guinea. AB - The band 3 deletion for southeast Asian ovalocytosis (SAO) occurs commonly in southeast Asia and the western Pacific. Southeast Asian ovalocytosis is associated with protection against cerebral malaria in children and therefore could reduce sequestration of erythrocytes parasitized by Plasmodium falciparum in the brain microvasculature. Sequestration of parasitized erythrocytes in the placenta accounts for much of the pathology of malaria during pregnancy. Therefore, we investigated the effect of SAO on malaria during pregnancy in the malaria-hyperendemic north coastal region of Papua New Guinea. The frequency of SAO in 927 women attending hospital for delivery was 8.7% (95% confidence interval = 6.9-10.5). Markers of fertility, the frequency of miscarriages and stillbirths, maternal anemia, placental and peripheral malaria at delivery, and birth weight were similar in women with and without SAO. In summary, although we can not exclude an interaction between SAO and malaria during pregnancy, we found no evidence that it provided a clinical benefit in this population. PMID- 17426162 TI - Caregivers' acceptance of using artesunate suppositories for treating childhood malaria in Papua New Guinea. AB - Community-based interventions using artemisinin-derived suppositories may potentially reduce malaria-related childhood mortality. However, their sociocultural acceptability is unknown in Papua New Guinea and a formal examination of caregiver's attitudes to rectal administration was needed to inform effective deployment strategies. Caregivers (n = 131) of children with uncomplicated malaria were questioned on their prior experience with, and attitudes to, rectal administration and then offered artesunate suppositories as treatment of their child. The 29% who refused this alternative were further questioned to determine their reasons for this refusal. Lack of spousal approval and fear of side effects were the most common reasons for refusal. Sixty-six percent of caregivers agreed to self-administer suppositories, which were perceived as effective (99%), safe (96%), and fast-acting (91%), but problematic to administer to a struggling child (56%). Shame, embarrassment, and hygiene were not significant concerns. Acceptability of rectal administration should be relatively high in Papua New Guinea. However, deployment must be accompanied by health education that addresses the practical aspects of administration, is appropriate for the illiterate, and is directed at fathers as well as mothers. PMID- 17426163 TI - Pfmdr1 and in vivo resistance to artesunate-mefloquine in falciparum malaria on the Cambodian-Thai border. AB - Artemisinin combination therapies (ACTs) have recently been adopted as first-line therapy for Plasmodium falciparum infections in most malaria-endemic countries. In this study, we estimated the association between artesunate-mefloquine therapy failure and genetic changes in the putative transporter, pfmdr1. Blood samples were acquired from 80 patients enrolled in an 2004 in vivo efficacy study in Pailin, Cambodia, and genotyped for pfmdr1 copy number and haplotype. Having parasites with three or more copies of pfmdr1 before treatment was strongly associated with recrudescence (hazard ratio [HR] = 8.30; 95% CI: 2.60-26.43). This relationship was maintained when controlling for initial parasite density and hematocrit (HR = 7.91; 95% CI: 2.38-26.29). Artesunate-mefloquine treatment selected for increased pfmdr1 copy number, because isolates from recurrent episodes had higher copy numbers than the paired enrollment samples (Wilcoxon rank test, P = 0.040). pfmdr1 copy number should be evaluated further as a surveillance tool for artesunate-mefloquine resistance in Cambodia. PMID- 17426164 TI - Disseminated intravascular coagulation complicated by peripheral gangrene in a rhesus macaque (Macaca mulatta) experimentally infected with Plasmodium coatneyi. AB - We report the first case of disseminated intravascular coagulation (DIC) complicated by peripheral gangrene induced by Plasmodium coatneyi in rhesus monkeys. Ten days after experimental challenge, numerous petechiae were noted over the trunk and extremities, with polychromasia, severe anemia, thrombocytopenia, and moderate parasitemia. These changes were accompanied by elevated serum activity of blood urea nitrogen, creatinine, transaminases, and creatinine phosphokinase. The animal received intravenous fluid support, artemether, and blood transfusion. Three days after treatment, the platelet counts returned to normal, and parasitemia was abated. However, several areas of skin discoloration with gangrenous tissue in the hands and the tail were observed. Coagulation profile showed elevated D-dimers and elevated levels of fibrinogen/fibrin degradation products with low levels of protein S functional activity. DIC with peripheral gangrene is very rare in Plasmodium-infected individuals. Our results indicate that the experimental model of P. coatneyi infection of rhesus monkeys is important for studies of malarial anemia and coagulopathy. PMID- 17426165 TI - Efficacy of atovaquone-proguanil for treatment of acute multidrug-resistant Plasmodium falciparum malaria in Thailand. AB - A combination of atovaquone-proguanil (Malarone); GlaxoSmithKline, Research Triangle Park, NC) was previously shown to be highly effective in the treatment of uncomplicated Plasmodium falciparum malaria. However, there are only limited recent efficacy data, particularly from regions of multidrug resistance. In this study, we examined the efficacy of atovaquone-proguanil for the treatment of uncomplicated P. falciparum malaria on the Thailand-Myanmar border. Patients were given directly observed atovaquone-proguanil (1,000 mg/400 mg) once a day for three days and followed-up for four weeks in a non-transmission area. Of 140 eligible patients enrolled in this open-label study, 97.8% (95% confidence interval = 95.4-100%) responded to therapy and remained clear of parasitemia at follow-up. Mean parasite clearance time was 41.9 hours and mean fever clearance time was 37.1 hours. On the basis of genotyping, three cases of treatment failure were identified (1 RIII and 2 RI). These data indicate that atovaquone-proguanil remains highly efficacious for the treatment of multidrug-resistant P. falciparum malaria in Thailand. PMID- 17426166 TI - Daily survival rates and dispersal of Aedes aegypti females in Rio de Janeiro, Brazil. AB - Daily survival rates, life expectancy, dispersal, and parity are important components of vectorial capacity of Aedes aegypti. These parameters were estimated for mosquito populations from a slum and a suburban district in Rio de Janeiro, during the wet and dry seasons in 2005. In each mark-release-recapture experiment, three cohorts of dust-marked Ae. aegypti females were released. Recaptures were carried out daily in randomly selected houses, using backpack aspirators, adult traps, and sticky ovitraps. Recapture varied between 6.81% and 14.26%. Daily survival was estimated by fitting two alternative models: exponential and nonlinear models with correction for the removal of individuals. Slum area presented higher survival and parity rates (68.5%). Dispersal rates were higher in the suburban area, where a maximum dispersal of 363 m was observed. Results suggest intense risk of dengue epidemic, particularly in the urban area. PMID- 17426167 TI - Infestation of Wistar rats with Tunga penetrans in different microenvironments. AB - Tungiasis is a zoonotic ectoparasitosis that causes considerable morbidity in affected populations. The type of microenvironment that facilitates infestation of hosts by Tunga penetrans has not been investigated. In this study, we exposed 30 laboratory-raised Wistar rats, a suitable model for the infestation, at six different places characterized by different microenvironments in a hyperendemic fishing village in northeastern Brazil. During a period of two weeks, the animals were monitored and the number of embedded fleas was documented. The number of lesions varied considerably according to the microenvironment and was highest in a cage placed at the far end of a compound of a household affected by tungiasis. No penetration was observed inside houses. Results indicate that in this endemic area transmission of T. penetrans seems to occur mainly outdoors. PMID- 17426168 TI - Macrophage killing of Leishmania amazonensis amastigotes requires both nitric oxide and superoxide. AB - The requirements for effective and efficient intracellular killing of Leishmania amazonensis by activated macrophages are unknown. Despite resistance to the arginase inhibitor LOHA by intracellular L. amazonensis amastigotes, enhanced replication did not account for the relative resistance of this parasite to macrophage activation. Herein we report that the presence of both superoxide and nitric oxide is necessary for efficient killing of L. amazonensis amastigotes within LPS/IFN-gamma-activated bone marrow-derived macrophages generated from C3H mice. Addition of an extracellular signal-regulated kinase (ERK) inhibitor to L. amazonensis-infected macrophages increased the ability of these activated macrophages to kill L. amazonensis amastigotes. This enhanced macrophage killing through addition of ERK inhibitor was abrogated by inhibition of superoxide or iNOS, whereas inhibiting superoxide had no effect on the killing of L. major. These results suggest that ERK activation may modulate effective macrophage killing, leading to the ability of L. amazonensis to resist elimination within activated macrophages. PMID- 17426169 TI - Cross-sectional serosurvey of feline leishmaniasis in ecoregions around the Northwestern Mediterranean. AB - A cross-sectional serosurvey using Leishmania infantum ELISA was performed on 445 cats living in ecoregions around the Northwestern Mediterranean basin; 58 cats from an area of the US where leishmaniasis is not endemic were used as negative controls. ELISA results were further confirmed in 69 cats by Western blot (WB). Finally, 76 of them were also tested for FeLV and FIV. Seroprevalence by ELISA prot A was 6.29%, and that by ELISA-IgG was 5.25%. Positive cat sera recognized patterns of polypeptides in WB, including L. infantum-specific antigenic fractions. There was no association with retroviruses. Leishmania-specific antibodies are prevalent in cats living in ecoregions around the Northwestern Mediterranean basin; thus, leishmaniasis must be included in the differential diagnosis of diseases in cats living in these ecoregions. Their role as peridomestic reservoirs for L. infantum needs further characterization, but it could be hypothesized that the cat is a secondary reservoir host, rather than an accidental one. PMID- 17426170 TI - Characterization of natural antimony resistance in Leishmania donovani isolates. AB - Clinical resistance to pentavalent antimonial compounds has long been recognized as a major problem in the treatment of visceral leishmaniasis in India. However, mechanisms of natural resistance are unclear. In this study, we observed that Leishmania donovani clinical isolates not responsive to sodium stibogluconate showed resistance to antimony treatment in both in vitro and in vivo laboratory conditions. The resistant isolates have increased levels of intracellular thiols. This increase in thiol levels was not mediated by the amplification of gamma glutamylcysteine synthetase, but was accompanied by amplification of trypanothione reductase and an intracellular ATP-binding cassette transporter gene MRPA. The resistance of parasites to antimony could be reversed by the glutathione biosynthesis-specific inhibitor, buthionine sulfoximine, which resulted in increased drug susceptibility. These results suggest the possible role of thiols and MRPA in antimony resistance in field isolates. PMID- 17426171 TI - Utility of lymph node aspiration in the diagnosis of visceral leishmaniasis in Sudan. AB - We evaluated lymph node aspiration (LNA) as a simple diagnostic procedure for visceral leishmaniasis (VL). Lymph node aspiration was compared with the direct agglutination test (DAT) using a diagnostic titer > or = 1:6,400 in 7,880 suspected VL patients in eastern Sudan. Compared with DAT, LNA had a sensitivity of 65.1% (95% confidence interval = 63.5-66.6%). Parasite density in LNA correlated strongly with DAT titers (P < 0.0001), and low parasite density accounted for 78.1% of positive LNA results with DAT titers < 1:6,400 (n = 782). Risk factors predictive of a positive LNA result were an age of 1-29 years, male sex, a hemoglobin level < 10.0 g/dL, a DAT titer > or = 1:800, and a location with a higher prevalence of VL. Lymph node and splenic aspirations were similarly accurate as tests of cure after treatment of 50 VL patients in southern Sudan. Pre-treatment LNA results were negative in 20 cases of severe post kala-azar dermal leishmaniasis. PMID- 17426172 TI - Molecular evidence of Babesia equi transmission in Haemaphysalis longicornis. AB - We studied the tick, Haemaphysalis longicornis, to determine the possibility of both transovarial and transstadial transmission of Babesia equi. We also studied the usefulness of the needle injection method for pathogenic tick-transmitted organisms including Babesia parasites. Erythrocytes infected with B. equi were injected into the midgut of engorged adults or nymphs using a hypodermic needle passed through the integument. DNA of B. equi in ticks was detected using nested polymerase chain reaction (PCR). B. equi DNA was present in adults, eggs, and larvae, indicating that transovarial transmission occurred. B. equi DNA was present in adults that developed from infected nymphs, and the B. equi antigen was present in their salivary glands, indicating that transstadial transmission occurred. These findings suggest that H. longicornis may play a role in the transmission of B. equi. PMID- 17426173 TI - Evaluation of brucellosis by PCR and persistence after treatment in patients returning to the hospital for follow-up. AB - Polymerase chain reaction (PCR) was applied to confirm the diagnosis of brucellosis and to study its clearance in response to the standard treatment regimen with doxycycline and rifampin at hospitals in Callao and Lima, Peru. The PCR confirmed the diagnosis in 23 (91.7%) patients with brucellosis including 12 culture-confirmed cases. For patients treated at the hospital in Callao, PCR was positive for all samples collected during and at the conclusion of treatment and for 76.9% of follow-up samples collected on average 15.9 weeks after completion of treatment. For patients treated at the hospital in Lima, PCR tests were positive for 81.8% of samples collected during treatment, for 33.3% of samples collected at the conclusion of treatment, and for > or = 50% of samples collected at first, second, and third post-treatment follow-up. Thus, Brucella DNA may persist in the serum weeks to months after completion of the standard treatment regimen. PMID- 17426174 TI - High prevalence of HIV and syphilis in a remote native community of the Peruvian Amazon. AB - Little data are available on how HIV and other sexually transmitted infections (STIs) affect indigenous people in Latin America, including Peru. We conducted a sero-epidemiologic survey of HIV infection and syphilis in a native community, the Chayahuita, an indigenous population in the Amazon region of Peru. The seroprevalences of HIV and syphilis in adults were 7.5% (6 of 80) and 6.3% (5 of 80), respectively. None of the participants had ever used a condom. Male to male sexual behavior was common. At the current levels of HIV prevalence, there is the risk of a negative impact on the survival of the Chayahuita ethnic group as a whole. The outcomes of this study highlight the need for urgent medical and anthropologic approaches to stop HIV transmission in indigenous Amazonian communities. PMID- 17426175 TI - Geographical distribution and risk factors associated with enteric diseases in Vietnam. AB - In Vietnam, shigellosis, typhoid fever, and cholera are important enteric diseases. To determine their magnitude and geographical distribution, and explore associated risk factors, we examined national surveillance data from 1991 to 2001 and potential ecological determinants. Average annual incidence rates were calculated and mapped for each province. Bivariate and multiple regression analyses were used to explore associations with selected environmental and human risk factors. Overall, shigellosis rates per 100,000 population (median, 41; mean, 70) were higher and more widespread than rates for typhoid fever (median, 7; mean, 23) and cholera (median, 0.3; mean, 2.7). Shigellosis was highest in the Central Highlands and was significantly associated with rainfall and urban poverty; typhoid fever prevailed in the Mekong River Delta and was most associated with vapor pressure and river/stream drinking water; and cholera predominated along the Central Coastal regions and correlated positively with rainfall and public well drinking water. The distinct geographical patterns of each disease appear to be driven by a combination of different ecological factors. PMID- 17426176 TI - Multiplex real-time PCR assay for detection of Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. AB - Entamoeba histolytica, Giardia intestinalis, and Cryptosporidium spp. are not only three of the most important and common diarrhea-causing parasitic protozoa, but they often have similar clinical presentations. Microscopic diagnosis of these parasites is neither sensitive nor specific. Recently, more specific and sensitive alternative molecular methods (polymerase chain reaction [PCR] and antigen detection tests) have been introduced for all three of these parasitic infections. The use of these molecular diagnostic tests in routine diagnostic laboratories is still limited. In this study, we developed a multiplex real-time PCR assay for the simultaneous detection of E. histolytica, G. intestinalis, and Cryptosporidium spp. in one reaction using species-specific probes. This assay was evaluated using clinical specimens and was found to be quite sensitive and specific. The reagents used in this multiplex PCR assay can also be used for detection of these parasites individually. The use of this real-time PCR multiplex assay in developing countries at present will have limited scope for routine diagnosis because the cost will be high for a single test, although in the developed world, the test could see immediate application. PMID- 17426177 TI - Diarrheal disease among HIV-infected adults in Karnataka, India: evaluation of risk factors and etiology. AB - The objectives of this study were to evaluate characteristics associated with diarrhea, the effect of trimethoprim-sulfamethoxazole (TMP/SMX) prophylaxis on diarrhea, the response to treatment with ciprofloxacin and tinidazole (Cipro-TZ), and presence of enteric pathogens. Adults infected with human immunodeficiency virus with and without diarrhea served as cases and controls, respectively. Participants provided a medical history and underwent a physical examination. Blood was collected for CD4 cell counts and stool for culture. Cases were treated with Cipro-TZ. Factors associated with a risk of diarrhea included crowded living and no toilet (all P < 0.05). Protective variables (P < 0.05) included a CD4 count greater than 200 cells/mm(3) and TMP/SMX prophylaxis. Cases were more likely to have a pathogen identified (P = 0.05). Eighty-six percent of the cases responded to treatment. Important risk factors for diarrhea were identified. Protection by TMP/SMX reinforces the importance of prophylaxis. These data suggest that treatment with an antibiotic and anti-parasitic medication may be effective. PMID- 17426178 TI - Two cases of rarely recognized infection with Entamoeba moshkovskii. AB - We report two cases (a 36-year-old woman and 2-year-old girl) infected with Entamoeba moshkovskii in Turkey. Entamoeba moshkovskii was identified and distinguished from the morphologically identical parasites E. histolytica and E. dispar by a nested polymerase chain reaction and restriction fragment length polymorphism analysis of the small subunit ribosomal DNA gene. PMID- 17426179 TI - A comparison of methods for detecting the eggs of Ascaris, Trichuris, and hookworm in infant stool, and the epidemiology of infection in Zanzibari infants. AB - This study compared five methods for detecting the eggs of the human parasitic geohelminths Ascaris, Trichuris, and hookworm in infant stool, and describes the epidemiology of infection in infants from a parasite-endemic area. A total of 424 infants 5-11 months old were enrolled from three villages on Pemba Island, Zanzibar. Methods used included the Kato-Katz technique, formol ethyl acetate sedimentation, modified formol ethyl acetate sedimentation, modified Wisconsin floatation, and simple gravity sedimentation. Of methods used alone, Wisconsin floatation and simple gravity sedimentation each provided the highest sensitivity for detecting eggs of these three geohelminths (89.6%). Of methods used in combination, the Kato-Katz technique/simple gravity sedimentation and Wisconsin floatation/simple gravity sedimentation each provided the highest sensitivity (99.0%). Prevalence of geohelminth infection was 26.5%. Between five and nine months of age the mean prevalence was 9.4%, while at 10 and 11 months of age the mean prevalence was 43.4%. Village prevalence varied from 3.6% to 43.8%. Infant geohelminth infection can occur at a high prevalence, and what method is best depends on research specifics. PMID- 17426180 TI - Low efficacy of mebendazole against hookworm in Vietnam: two randomized controlled trials. AB - Vietnam is participating in a global de-worming effort that aims to treat 650 million school children regularly by 2010. The treatment used in Vietnam is single dose oral mebendazole (Phardazone) 500 mg. We tested the efficacy of single dose mebendazole 500 mg in the therapy of hookworm infection in a randomized double-blind placebo-controlled trial among 271 Vietnamese schoolchildren. The treatment efficacy of single dose mebendazole in children did not differ significantly from placebo, with a reduction in mean eggs per gram of feces relative to placebo of 31% (95% CI -9 to 56%, P = 0.1). In light of these findings we then carried out a similar randomized trial comparing triple dose mebendazole, single dose albendazole, and triple dose albendazole against placebo in 209 adults in the same area. The estimated reduction in mean post-treatment eggs per gram of feces relative to placebo was 63% (95% CI 30-81%) for triple mebendazole, 75% (47-88%) for single albendazole, and 88% (58-97%) for triple albendazole. Our results suggest that single dose oral mebendazole has low efficacy against hookworm infection in Vietnam, and that it should be replaced by albendazole. These findings are of major public health relevance given the opportunity costs of treating entire populations with ineffective therapies. We recommend that efficacy of anti-helminth therapies is pilot tested before implementation of national gut worm control programs. PMID- 17426181 TI - Thrombocytopenia associated with dengue hemorrhagic fever responds to intravenous administration of anti-D (Rh(0)-D) immune globulin. AB - Severe thrombocytopenia and increased vascular permeability are two major characteristics of dengue hemorrhagic fever (DHF). An immune mechanism of thrombocytopenia due to increased platelet destruction appears to be operative in patients with DHF (see Saito et al., 2004, Clin Exp Immunol 138: 299-303; Mitrakul, 1979, Am J Trop Med Hyg 26: 975-984; and Boonpucknavig, 1979, Am J Trop Med Hyg 28: 881-884). The interim data of two randomized placebo controlled trials in patients (N = 47) meeting WHO criteria for dengue hemorrhagic fever (DHF) with severe thrombocytopenia (platelets < or = 50,000/mm(3)) reveal that the increase in platelet count with anti-D immune globulin (WinRho SDF), 50 microg/kg (250 IU/kg) intravenously is more brisk than the placebo group. The mean maximum platelet count of the anti-D-treated group at 48 hours was 91,500/mm(3) compared with 69,333/mm(3) in the placebo group. 75% of the anti-D treated group demonstrated an increase of platelet counts > or = 20,000 compared with only 58% in the placebo group. These data suggest that treatment of severe thrombocytopenia accompanying DHF with anti-D may be a useful and safe therapeutic option. PMID- 17426182 TI - Induction of bivalent immune responses by expression of dengue virus type 1 and type 2 antigens from a single complex adenoviral vector. AB - There are approximately 100 million new cases of dengue (DEN) virus infection each year. Infection can result in illness ranging from a mild fever to hemorrhaging, shock, or even death. There are four serotypes of dengue virus (DEN1-4), and immunity to one serotype does not cross protect from infection with other serotypes. Currently there are no approved vaccines for dengue fever. In this report, we describe the construction of a bivalent dengue virus vaccine using a complex recombinant adenovirus approach to express multiple genes of DEN1 and DEN2 serotypes. In vaccinated mice, this vector induced humoral immune responses against all four dengue serotypes as measured by enzyme-linked immunosorbent assay. However, the neutralizing antibody responses were specific for DEN1 and DEN2 serotypes. Expansion of this vaccine development platform towards the DEN3 and DEN4 serotypes can lead towards the development of an adenovirus-based tetravalent dengue vaccine. PMID- 17426183 TI - Difficulty in diagnosis and treatment of dengue hemorrhagic fever in patients with chronic renal failure: report of three cases of mortality. AB - In 2002, an outbreak of dengue fever (DF) in Taiwan caused mortality in some patients with chronic renal failure (CRF). We report three cases of CRF who died of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) despite an intensive care process. The difficulty in diagnosis and the treatment dilemma attributable to ambiguity in symptoms and signs between CRF and DF may delay the diagnosis, contributing to a high risk of mortality. The narrow window of fluid tolerance in patients with CRF further hampers the success of resuscitation in DHF and DSS. Continuous venous to venous hemodialysis (CVVHD) is helpful in managing a condition with unstable hemodynamics. However, to decrease mortality, the physician must pay great attention to ensure an early awareness of DHF/DSS and deliver a prompt and aggressive treatment of patients of dengue viral infections with chronic renal failure. PMID- 17426184 TI - Monkeypox zoonotic associations: insights from laboratory evaluation of animals associated with the multi-state US outbreak. AB - At the onset of the 2003 US monkeypox outbreak, virologic data were unavailable regarding which animal species were involved with virus importation and/or subsequent transmission to humans and whether there was a risk for establishment of zoonotic monkeypox in North America. Similarly, it was unclear which specimens would be best for virus testing. Monkeypox DNA was detected in at least 33 animals, and virus was cultured from 22. Virus-positive animals included three African species associated with the importation event (giant pouched rats, Cricetomys spp.; rope squirrels, Funisciuris sp.; and dormice, Graphiuris sp.). Virologic evidence from North American prairie dogs (Cynomys sp.) was concordant with their suspected roles as vectors for human monkeypox. Multiple tissues were found suitable for DNA detection and/or virus isolation. These data extend the potential host range for monkeypox virus infection and supports concern regarding the potential for establishment in novel reservoir species and ecosystems. PMID- 17426185 TI - Efficacy of the antipoxvirus compound ST-246 for treatment of severe orthopoxvirus infection. AB - Efficacy of the new antipoxvirus compound ST-246 was evaluated as treatment of monkeypox (MPX) virus infection in a ground squirrel model of the disease. Ground squirrels were given a lethal dose of MPX virus and were then treated orally at various times post-inoculation (pi) with 100 mg/kg/day of ST-246. Morbidity and mortality, clinical laboratory results, viral load, and pathology of placebo and treatment groups were compared. All animals that started treatment with ST-246 on days 0, 1, 2, and 3 pi survived lethal challenge with MPX virus; 67% of animals treated on day 4 pi also survived. In contrast, 100% of the placebo group died. Most of the ST-246-treated animals showed no evidence of clinical disease or alteration of baseline clinical laboratory values and had minimal histopathologic changes. These results suggest that ST-246 is a promising candidate for early treatment of severe orthopoxvirus infection. PMID- 17426186 TI - Recombinant alphaviruses are safe and useful serological diagnostic tools. AB - Serological assays for diagnosis of Venezuelan equine encephalitis virus (VEEV) currently require bio-safety level 3 facilities and select agent certification to produce antigens, reference sera, or viral stocks. Rapid identification of VEEV infection is required to respond to human and equine outbreaks of encephalitis caused by that virus and can be useful for epidemiologic surveillance. Alphavirus (Sindbis)-based recombinant viruses that express VEEV structural proteins are attenuated in animal models, thus representing an alternative to the handling of virulent infectious virus. Virus and viral antigens from recombinant Sindbis/VEE constructs engineered to express structural proteins from multiple VEEV subtypes were evaluated as diagnostic reagents in VEEV-specific serological assays, e.g., plaque reduction neutralization test (PRNT), hemagglutination inhibition (HI) assay, and complement fixation (CF) test. Chimeric viruses were produced efficiently in cell culture and were as effective as the parental virus for identifying infection of humans, horses, and rodents in these serological assays. PMID- 17426187 TI - Fox squirrel (Sciurus niger) associations with West Nile virus. AB - Tree squirrels (Sciurus spp.) have been recently shown to be commonly exposed to West Nile virus (WNV). Many characteristics of WNV infections in tree squirrels are unknown. To better understand WNV associations in fox squirrels (S. niger), we conducted mark-recapture sampling (N = 72) and radio telemetry to study the longitudinal seroprevalence, seroconversions, and ectoparasites of these animals during 2005-2006 in northern Colorado. Five seroconversions were documented during this study. The majority of seroconversions occurred during the late summer/fall months. However, one seroconversion was documented over the time period of February to late March 2005. Fleas (Orchopeas howardi) were tested for WNV RNA using real-time PCR techniques. No WNV RNA positive fleas (N = 33) were detected. In addition, urine samples (N = 17) opportunistically collected from fox squirrels were negative for WNV RNA. Results indicate that seroconversions can be observed in fox squirrels during low WNV transmission years. PMID- 17426188 TI - Dhori virus (Orthomyxoviridae: Thogotovirus) infection in mice: a model of the pathogenesis of severe orthomyxovirus infection. AB - After intranasal, subcutaneous, or intraperitoneal infection with Dhori virus (DHOV), adult mice developed a fulminant and uniformly fatal illness with many of the clinical and pathologic findings seen in mice infected with H5N1 highly pathogenic avian influenza A virus. Histopathologic findings in lungs of DHOV infected mice consisted of hemorrhage, inflammation, and thickening of the interstitium and the alveolar septa and alveolar edema. Extra-pulmonary findings included hepatocellular necrosis and steatosis, widespread severe fibrinoid necrosis in lymphoid organs, marked lymphocyte loss and karyorrhexis, and neuronal degeneration in brain. Similar systemic histopathologic findings have been reported in the few fatal human H5N1 cases examined at autopsy. Because of the relationship of DHOV to the influenza viruses, its biosafety level 2 status, and its similar pathology in mice, the DHOV-mouse model may offer a low-cost, relatively safe, and realistic animal model for studies on the pathogenesis and management of H5N1 virus infection. PMID- 17426189 TI - Perceptions of smoking and nonsmoking peers: the value of smoker and nonsmoker prototypes in predicting smoking onset and regular smoking among adolescents. AB - Adolescents' perceptions of persons their age who smoke cigarettes (also known as prototypes of smoking peers) play a critical role in an adolescent's decision to start smoking. However, adolescents' perceptions of their peers who do not smoke (prototypes of nonsmoking peers) could be implicated in adolescents' smoking decisions as well. In the present study, the authors examined the additional role of nonsmoker prototypes in adolescents' smoking onset and regular smoking. At seven high schools, Dutch students (n = 1,035) between the ages of 12 and 15 years who were attending the eighth grade provided self-reported data on their smoker and nonsmoker prototypes and smoking behavior during a baseline and 6 month follow-up measurement. Logistic regression analyses showed that both smoker and nonsmoker prototypes assessed at Time 1 predicted smoking onset by Time 2 among nonsmoking adolescents. However, only nonsmoker prototypes predicted regular smoking among adolescents who smoked occasionally at baseline. PMID- 17426190 TI - Glucocorticoids for ARDS: Just Do It! PMID- 17426191 TI - Atrial septostomy: why we still need it. PMID- 17426192 TI - The staging system for non-small cell lung cancer: time for an overhaul? PMID- 17426193 TI - Clarion call for trials assessing "cardiopulmonary" agents to reduce morbidity and mortality in inflammatory lung diseases. PMID- 17426194 TI - Comet tails in high-altitude pulmonary edema: diagnostic portent or streak in the sky? PMID- 17426195 TI - Methylprednisolone infusion in early severe ARDS: results of a randomized controlled trial. AB - OBJECTIVE: To determine the effects of low-dose prolonged methylprednisolone infusion on lung function in patients with early severe ARDS. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: ICUs of five hospitals in Memphis. PARTICIPANTS: Ninety-one patients with severe early ARDS (/= 1,200 pg/mL, BNP had a specificity of 92% for CPE. Higher levels of BNP were associated with a decreased odds for ARDS (odds ratio, 0.4 per log increase; p = 0.007) after adjustment for age, history of CHF, and right atrial pressure. BNP was associated with in-hospital mortality (p = 0.03) irrespective of the final diagnosis and independent of APACHE (acute physiology and chronic health evaluation) II score. CONCLUSION: In ICU patients with hypoxemic respiratory failure, BNP appears useful in excluding CPE and identifying patients with a high probability of ARDS, and was associated with mortality in patients with both ARDS and CPE. Larger studies are necessary to validate these findings. PMID- 17426197 TI - Outcome of patients with small-vessel vasculitis admitted to a medical ICU. AB - PURPOSES: This study aims to describe the clinical course and prognostic factors of patients with small-vessel vasculitis admitted to a medical ICU. METHODS: We reviewed the clinical records of 38 patients with small-vessel vasculitis admitted consecutively to the ICU between January 1997 and May 2004. The APACHE (acute physiology and chronic health evaluation) III prognostic system was used to determine the severity of illness on the first ICU day; the sequential organ failure assessment (SOFA) score was used to measure organ dysfunction, and the Birmingham vasculitis activity score for Wegener granulomatosis (BVAS/WG) was used to assess vasculitis activity. Outcome measures were the 28-day mortality and ICU length of stay. RESULTS: Nineteen patients (50%) had Wegener granulomatosis, 16 patients (42%) had microscopic polyangiitis, 2 patients had CNS vasculitis, and 1 patient had Churg-Strauss syndrome. Reasons for ICU admission included alveolar hemorrhage in 14 patients (37%), sepsis in 5 patients (13%), seizures in 3 patients (8%), and pneumonia in 2 patients (5%). The median ICU length of stay was 4.0 days (interquartile range, 2.0 to 6.0 days). The APACHE III score was lower in survivors than nonsurvivors (p = 0.010). The predicted hospital mortality was 54% for nonsurvivors and 21% for survivors (p = 0.0038). The mean SOFA score was 11.6 (SD, 2.6) in nonsurvivors, compared to 6.9 (SD, 2.4) in survivors (p = 0.0004). Mean BVAS/WG scores were 8.6 (SD, 3.6) in nonsurvivors and 4.7 (SD, 4.6) in survivors (p = 0.0889). Twenty-six percent of the patients received invasive mechanical ventilation, and 33% underwent dialysis. The 28-day and 1-year mortality rates were 11% and 29%, respectively. CONCLUSIONS: The mortality of patients with small-vessel vasculitis admitted to the ICU is lower than predicted, and alveolar hemorrhage is the most common reason for ICU admission. PMID- 17426198 TI - Atrial septostomy in treatment of end-stage right heart failure in patients with pulmonary hypertension. AB - BACKGROUND: Right ventricular (RV) failure is the main cause of death in patients with pulmonary hypertension (PH). Balloon atrial septostomy (BAS) is believed to relieve symptoms of PH by increasing systemic flow and reducing RV preload. METHODS: Fourteen BAS procedures were performed in 11 patients (5 men and 6 women; mean [+/- SD] age, 33 +/- 12 years) with RV failure in the course of PH that was refractory to conventional treatment. BAS consisted of a puncture of the interatrial septum and subsequent dilatations with balloons of increasing diameter in a step-by-step manner. RESULTS: After BAS, the mean oxygen saturation of aortic blood decreased (before, 93 +/- 4%; after, 84 +/- 4%; p = 0.001), while mean cardiac index increased (before, 1.54 +/- 0.34 L/min/m(2); after, 1.78 +/- 0.35 L/min/m(2); p = 0.001), resulting in a positive trend for mean systemic oxygen transport (before, 270 +/- 64 mL/min; after, 286 +/- 81 mL/min; p = 0.08). Pulmonary vascular resistance (PVR) slightly increased immediately after the procedure, and this rise inversely correlated with mixed venous blood partial oxygen pressure both before BAS (r = -0.69; p = 0.009) and after BAS (r = -0.64; p = 0.018). Mean functional class improved from 3.2 +/- 0.4 to 2.6 +/- 0.7 (p = 0.03) after 1 month. At follow-up (mean time to follow-up, 8.1 +/- 6.2 months; range, 0.8 to 20.2 months), seven patients died and two underwent lung transplantation. There was no difference in the survival rate compared to that obtained from National Institutes of Health equation. A significant size reduction in the created defect was observed in six patients, requiring repeat BAS procedures in three cases. CONCLUSIONS: The current BAS technique improves cardiac index and functional class without significant periprocedural complications, except for a transient increase in PVR related to acute desaturation of mixed venous blood. At long-term follow-up, a high incidence of spontaneous decrease in orifice size has been observed. PMID- 17426199 TI - Long-term outcome in a patient with pulmonary hypertension and hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) may be associated with pulmonary hypertension (PH). In the context that little attention has been given to long term follow-up of such individuals, we report a patient with PH associated with HHT with special attention to clinical features and long-term response to therapy. To our knowledge, this case represents only the second with a 10-year follow-up reported and demonstrates that aggressive therapy can lead to long-term improvement in clinical parameters and survival. PMID- 17426200 TI - Pericardial abnormalities predict the presence of echocardiographically defined pulmonary arterial hypertension in systemic sclerosis-related interstitial lung disease. AB - OBJECTIVES: To determine the prevalence and significance of pericardial abnormalities in systemic sclerosis (SSc)-related interstitial lung disease (ILD). METHODS: Retrospective study of 41 subjects with SSc-related ILD who underwent evaluation including thoracic high-resolution CT (HRCT) imaging, transthoracic echocardiography (TTE), and pulmonary function testing. HRCT review evaluated the pericardium for the presence of pericardial effusion (PEf), thickness of the anterior pericardial recess (APR) [abnormal defined as > 10 mm], and pericardial thickening as calculated by total pericardial score (TPS) [abnormal defined as > 8 mm]. Pulmonary arterial hypertension (PAH) was defined as a pulmonary artery pressure > 35 mm Hg estimated by TTE. RESULTS: Fifty-nine percent had an abnormal pericardium, 49% had a PEf, 56% had an abnormal APR, and 49% had an abnormal TPS. An abnormal pericardium was more common in men than women. Subjects with and without pericardial abnormalities were otherwise similar with respect to age, SSc classification, autoantibodies, ILD radiographic pattern, and presence of esophageal dilation. Both groups had similar median percentage of predicted total lung capacity, percentage of predicted FVC, percentage of predicted FEV(1), and percentage of predicted diffusion capacity of the lung for carbon monoxide. Subjects with pericardial abnormalities were more likely to have coexistent PAH (35% vs 75%; p = 0.02) and a higher median right ventricular systolic pressure (31 mm Hg vs 44 mm Hg; p = 0.03). Multiple logistic regression revealed that TPS was the best individual predictor of the presence of TTE-defined PAH. CONCLUSIONS: In patients with SSc-related ILD, pericardial abnormalities are commonly seen on HRCT, and their presence is strongly associated with echocardiographically defined PAH, with abnormal TPS as the best individual predictor. PMID- 17426201 TI - Significance of extranodal extension of regional lymph nodes in surgically resected non-small cell lung cancer. AB - STUDY OBJECTIVES: Regional lymph node (LN) involvement affects the prognosis of patients with surgically resected non-small cell lung cancer (NSCLC). The significance of extranodal extension in these groups of patients was prospectively studied to determine its clinicopathologic relationships and its influence on patient survival. METHODS: A total of 199 NSCLC patients who were proved to have regional LN involvement after resection were included. Histologic examinations including tumor cell type, grade of differentiation, vascular invasion, regional LN metastasis emphasizing the number and station of LN involvement, the presence or absence of extranodal extension, and the immunohistochemistry of p53 expression were obtained. The relationships between extranodal extension and histologic type, grade of differentiation, vascular invasion, tumor size, pathologic stage, p53 expression, or patient survival were analyzed. RESULTS: Extranodal extension was significantly higher in women, adenocarcinoma, advanced stage, tumors with vascular invasion, or p53 overexpression. The total number and positive rate of resected LNs with extranodal extension were significantly correlated with advanced stage, tumors with vascular invasion, or p53 overexpression. By multivariate analysis of survival, the presence or total number of LNs with extranodal extension, tumor stage, and p53 expression were significant prognostic factors. The 5-year survival rate of stage IIIA patients without extranodal extension (30.4%) was significantly better than that of stage II patients with extranodal extension (16.8%). No survival difference between extranodal positive stage II and IIIA patients was noted. CONCLUSIONS: Extranodal extension of regional LNs is an important prognostic factor in patients with surgically resected NSCLC. PMID- 17426202 TI - Declining incidence rate of lung adenocarcinoma in the United States. AB - BACKGROUND: Adenocarcinoma of the lung (ADL) increased worldwide during the last half century. We now report that a continuous decline of ADL began in the United States in 1999. METHOD: Incidence rates of ADL and squamous cell carcinoma of the lung (SQL) from The Surveillance Epidemiology and End Results Program were reviewed for the 31-year period beginning in 1973. The low-tar cigarette (tar 200% in women. From 1999 through 2003, the rate declined 14% in men and 8% in women. An analysis of age-specific incidence rates of ADL according to birth cohort demonstrates that rates declined progressively among persons born after 1934 for both genders. The increase in low tar cigarette consumption did not precede the increase in ADL incidence rates, and the decline of ADL incidence after 1998 occurred without a preceding decline of low-tar cigarette consumption. CONCLUSION: Since 1999, the ADL incidence has declined. The temporal trend of ADL incidence may suggest that air pollution could be the possible determining cause for the trend. Increasing use of low-tar cigarettes in the United States and the decline in environmental tobacco smoke may be contributors but are less likely to be the driving force. PMID- 17426203 TI - Influenza and COPD mortality protection as pleiotropic, dose-dependent effects of statins. AB - BACKGROUND: Published data on antiinflammatory and immunomodulatory effects of statins suggest they may reduce mortality risks associated with an unchecked immune response to selected infections, including influenza and COPD. We assessed whether statin users had reduced mortality risks from these conditions. METHODS: We conducted a matched cohort study (n = 76,232) and two separate case-control studies (397 influenza and 207 COPD deaths) to evaluate whether statin therapy is associated with increased or decreased mortality risk and survival time using health-care encounter data for members of health maintenance organizations. For the cohort study, baseline illness risks from all causes prior to initiation of statin therapy were used to statistically adjust for the occurrence of outcomes after initiation of treatment. RESULTS: For moderate-dose (>/= 4 mg/d) statin users, this cohort study found statistically significant reduced odds ratios (ORs) of influenza/pneumonia death (OR, 0.60; 95% confidence interval [CI], 0.44 to 0.81) and COPD death (OR, 0.17; 95% CI, 0.07 to 0.42) and similarly reduced survival hazard ratios. Findings were confirmed with the case-control studies. Confounding factors not considered may explain some of the effects observed. CONCLUSIONS: This study found a dramatically reduced risk of COPD death and a significantly reduced risks of influenza death among moderate-dose statin users. PMID- 17426204 TI - Chest ultrasonography for the diagnosis and monitoring of high-altitude pulmonary edema. AB - BACKGROUND: The comet-tail technique of chest ultrasonography has been described for the diagnosis of cardiogenic pulmonary edema. This is the first report describing its use for the diagnosis and monitoring of high-altitude pulmonary edema (HAPE), the leading cause of death from altitude illness. METHODS: Eleven consecutive patients presenting to the Himalayan Rescue Association clinic in Pheriche, Nepal (4,240 m) with a clinical diagnosis of HAPE underwent one to three chest ultrasound examinations using the comet-tail technique to determine the presence of extravascular lung water (EVLW). Seven patients with no evidence of HAPE or other altitude illness served as control subjects. All examinations were read by a blinded observer. RESULTS: HAPE patients had higher comet-tail score (CTS) [mean +/- SD, 31 +/- 11 vs 0.86 +/- 0.83] and lower oxygen saturation (O(2)Sat) [61 +/- 9.2% vs 87 +/- 2.8%] than control subjects (p < 0.001 for both). Mean CTS was higher (35 +/- 11 vs 12 +/- 6.8, p < 0.001) and O(2)Sat was lower (60 +/- 11% vs 84 +/- 1.6%, p = 0.002) at hospital admission than at discharge for the HAPE patients with follow-up ultrasound examinations. Regression analysis showed CTS was predictive of O(2)Sat (p < 0.001), and for every 1-point increase in CTS O(2)Sat fell by 0.67% (95% confidence interval, 0.41 to 0.93%, p < 0.001). CONCLUSIONS: The comet-tail technique effectively recognizes and monitors the degree of pulmonary edema in HAPE. Reduction in CTS parallels improved oxygenation and clinical status in HAPE. The feasibility of this technique in remote locations and rapid correlation with changes in EVLW make it a valuable research tool. PMID- 17426205 TI - Diagnostic usefulness of fluorine-18-alpha-methyltyrosine positron emission tomography in combination with 18F-fluorodeoxyglucose in sarcoidosis patients. AB - OBJECTIVES: L-[3-(18)F]-alpha-methyltyrosine ((18)F-FMT) is an amino-acid tracer for positron emission tomography (PET) and is used for tumor detection because malignant tumor cells accumulate (18)F-FMT based on the increased expression of an amino-acid transporter. This study was conducted to investigate the usefulness of (18)F-FMT PET in combination with fluorine-18-fluorodeoxyglucose ((18)F-FDG) PET for the diagnosis of sarcoidosis in patients with suspected malignancy. SETTING: Twenty-four sarcoidosis patients with suspected malignancy underwent (18)F-FDG and (18)F-FMT PET. The study included 17 patients with extrapulmonary manifestation mimicking malignant disease (13 patients with systemic lymphadenopathy, 3 of them with concomitant hepatosplenic processes; 3 patients with hepatosplenic processes without concomitant lymphadenopathy; and 1 patient with multiple bone lesions), 3 patients with occurrence of bilateral hilar lymphadenopathy in cancer patients, and 4 patients with multiple nodules mimicking pulmonary metastasis. RESULTS: All patients showed increased uptake of (18)F-FDG and no increase in the accumulation of (18)F-FMT in their lymphadenopathy. Standardized uptake values (SUVs) of (18)F-FDG and (18)F-FMT were 5.01 +/- 2.15 and 0.77 +/- 0.24, respectively (mean +/- SD). All extranodal lesions such as liver, spleen, and bone were visually positive on (18)F-FDG PET and negative on (18)F-FMT PET. No neoplasm was confirmed in all patients. In a control group of patients with lung cancer, SUVs for (18)F-FDG and (18)F-FMT were 6.34 +/- 2.52 and 1.54 +/- 0.82, respectively. CONCLUSION: The uptake of (18)F FDG was positive in the sarcoid lesions, and therefore (18)F-FDG PET could not differentiate sarcoidosis from malignant disease. Use of (18)F-FMT PET in combination with (18)F-FDG PET may be the effective method to distinguish sarcoidosis from malignancy. PMID- 17426206 TI - Ability of low-dose helical CT to distinguish between benign and malignant noncalcified lung nodules. AB - STUDY OBJECTIVES: Low-dose helical CT scanning identifies early stage lung malignancies and also a large proportion of lung nodules of uncertain diagnostic and prognostic significance (ie, indeterminate nodules). The sensitivity, specificity, and predictive value of these indeterminate nodules detected by CT scanning as part of a lung cancer screening program is largely unknown. We therefore calculated the sensitivity, specificity, and predictive values of CT detected lung nodules that were followed up at least 18 months. DESIGN: Single arm screening trial with longitudinal follow-up. SETTING: Rural areas of United States, from 2000 to 2004. PARTICIPANTS: Former and current nuclear weapons workers, >/= 45 years old, including smokers and never-smokers, with variable exposure to occupational lung carcinogens. INTERVENTIONS: A total of 4,401 participants were CT scanned for lung cancer with an initial full chest low-dose CT scan, interval CT scans at 3, 6, and 12 months for indeterminate lung nodules (eg, nodules not immediately suspicious for lung cancer), and a 18-month, full chest, low-dose incidence CT scan. RESULTS: We achieved follow-up for a minimum of 18 months for > 95% of 807 participants with indeterminate or suspicious lung nodules. Only 3 of 727 indeterminate nodules were identified as being malignant during the subsequent 18 months. The radiologist's designation of a nodule as suspicious had a sensitivity of 84.2% and a specificity of 96.6%. Given a prior probability of lung cancer of 2.4%, positive and negative predictive values were 37.2% and 99.6%. Overall, we detected 33 primary lung cancers, including 19 stage I cancers, 5 stage II cancers, 7 stage III-IV cancers, and 3 limited-stage small cell cancers. CONCLUSIONS: Helical CT scanning detects many indeterminate nodules, but few are malignant. CT scanning has high sensitivity and specificity to detect early lung cancer. The problem of false-positive results in helical CT scanning is limited and can be rationally managed. Current CT follow-up recommendations are supported. PMID- 17426207 TI - Roles of angiopoietin-1 and angiopoietin-2 on airway microvascular permeability in asthmatic patients. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) increases microvascular permeability. Recently, considerable attention has been devoted to the physiologic roles of angiopoietin-1 and angiopoietin-2 as regulatory factors of VEGF. This study was designed to examine the roles of angiopoietin-1 and angiopoietin-2 in controlling airway microvascular permeability in asthma. METHODS: Levels of these angiogenic factors and airway vascular permeability index were examined in 30 asthmatics and 12 control subjects. After 2-week run-in period, all asthmatics were randomly assigned to receive fluticasone propionate (400 mug/d) or montelukast (10 mg) for 12 weeks. RESULTS: VEGF, angiopoietin-1, and angiopoietin-2 levels in induced sputum were significantly higher in asthmatics than in control subjects. We found an inverse correlation between angiopoietin-1 level and vascular permeability index in asthmatics, while there was a positive correlation between angiopoietin-2 level and that index. VEGF and angiopoietin-1 levels were significantly decreased after fluticasone therapy, while VEGF and angiopoietin-2 levels were significantly decreased after montelukast therapy. Although VEGF levels after treatment were different between two groups, vascular permeability index in the montelukast group was the same level as that in the fluticasone group. Moreover, improvement in vascular permeability index after fluticasone therapy was inversely correlated with decrease in angiopoietin-1 level, while that after montelukast therapy was positively correlated with decrease in angiopoietin-2 level. CONCLUSIONS: Angiopoietin-1 and angiopoietin-2 play complementary and coordinated roles in regulating microvascular permeability stimulated by VEGF in asthma. Combination of corticosteroids with leukotriene antagonists might effectively improve plasma leakage and provide a new strategy in treating bronchial asthma. PMID- 17426208 TI - Airway wall thickening in patients with cough variant asthma and nonasthmatic chronic cough. AB - BACKGROUND: Chronic cough, which may be of asthmatic or nonasthmatic origin, is an important clinical issue. Airway inflammation, and remodeling demonstrated by subbasement membrane thickening has been associated with cough variant asthma (CVA) as well as with nonasthmatic chronic cough (NAC). CT studies have shown airway wall thickening in patients with asthma who wheeze. We examined airway wall thickness by CT in adult patients with chronic cough and examined its pathophysiologic implication. METHODS: Nonsmoking, steroid-naive patients with CVA (n = 27), NAC (n = 26), and healthy control subjects (n = 15) were studied. Airway dimensions were assessed by a validated CT technique, in which we measured airway wall area (WA) corrected by body surface area (BSA), the ratio of WA to outer wall area (percentage of wall area [WA%]), absolute wall thickness (T)/ square root BSA, and airway luminal area/BSA of a segmental bronchus. Correlations between CT parameters and clinical indexes such as disease duration and cough sensitivity were examined. RESULTS: In patients with CVA, WA/BSA, WA%, and T/ square root BSA were all significantly greater than those in control subjects. In patients with NAC, WA/BSA and T/ square root BSA were significantly greater than in control subjects. The increase of WA/BSA and T/ square root BSA of NAC patients was less than that of CVA patients. In a subset of patients with NAC, WA% correlated with capsaicin cough sensitivity (n = 9, r = 0.75, p = 0.034). CONCLUSIONS: Walls of central airways are thickened in patients with CVA, and also to a lesser degree in patients with NAC. Airway wall thickening in NAC may be associated with cough hypersensitivity. PMID- 17426209 TI - Reduced intrathoracic blood volume and left and right ventricular dimensions in patients with severe emphysema: an MRI study. AB - BACKGROUND: Left ventricular (LV) filling is impaired in patients with severe emphysema manifesting in small end-diastolic dimensions. We hypothesized that the hyperinflated lungs of these patients with high intrinsic positive end-expiratory pressure will decrease intrathoracic blood volume (ITBV) and ventricular preload. We therefore measured ITBV, and LV and right ventricular (RV) dimensions and function using MRI techniques in patients with severe emphysema. METHODS: Patients with severe emphysema (n = 13) and matched healthy volunteers (n = 11) were included. The magnetic resonance (MR) examination consisted of three parts: (1) evaluation of RV and LV dimensions and function and interventricular septum curvature using cine MRI; (2) quantification of aortic flow using MR phase velocity mapping; and (3) calculation of the cardiopulmonary peak transit time (PTT) from the pulmonary artery to the ascending aorta using contrast-enhanced, time-resolved, two-dimensional MR angiography. RESULTS: There were no differences between the groups regarding age, height, or weight. In the emphysema patients, ITBV index (- 35%), LV end-diastolic volume index (LVEDVI) [- 21%], RV end diastolic volume index (- 20%), cardiac index (- 22%), and stroke volume index (SVI) [- 40%] were lower compared to control subjects. LV and RV end-systolic volumes, LV wall mass, septal curvature, and PTT did not differ between the groups. LVEDVI (r = 0.83) as well as SVI (r = 0.82) correlated closely to ITBV index. SVI correlated closely to LVEDVI (r = 0.84). CONCLUSIONS: LV and RV performance is impaired in patients with severe emphysema because of small end diastolic dimensions. One possible explanation for the decreased biventricular preload in these patients is intrathoracic hypovolemia caused by hyperinflated lungs. PMID- 17426210 TI - Copeptin, C-reactive protein, and procalcitonin as prognostic biomarkers in acute exacerbation of COPD. AB - BACKGROUND: A novel approach to estimate the severity of COPD exacerbation and predict its outcome is the use of biomarkers. We assessed circulating levels of copeptin, the precursor of vasopressin, C-reactive protein (CRP), and procalcitonin as potential prognostic parameters for in-hospital and long-term outcomes in patients with acute exacerbation of COPD (AECOPD) requiring hospitalization. METHODS: Data of 167 patients (mean age, 70 years; mean FEV(1), 39.9 +/- 16.9 of predicted [+/- SD]) presenting to the emergency department due to AECOPD were analyzed. Patients were evaluated based on clinical, laboratory, and lung function parameters on hospital admission, at 14 days, and at 6 months. RESULTS: Plasma levels of all three biomarkers were elevated during the acute exacerbation (p < 0.001), but levels at 14 days and 6 months were similar (p = not significant). CRP was significantly higher in patients presenting with Anthonisen type I exacerbation (p = 0.003). In contrast to CRP and procalcitonin, copeptin on hospital admission was associated with a prolonged hospital stay (p = 0.002) and long-term clinical failure (p < 0.0001). Only copeptin was predictive for long-term clinical failure independent of age, comorbidity, hypoxemia, and lung functional impairment in multivariate analysis (p = 0.005). The combination of copeptin and previous hospitalization for COPD increased the risk of poor outcome (p < 0.0001). Long-term clinical failure was observed in 11% of cases with copeptin < 40 pmol/L and no history of hospitalization, as compared to 73% of patients with copeptin >/= 40 pmol/L and a history of hospitalization (p < 0.0001). CONCLUSIONS: We suggest copeptin as a prognostic marker for short-term and long-term prognoses in patients with AECOPD requiring hospitalization. PMID- 17426211 TI - Effects of a walking aid in COPD patients receiving oxygen therapy. AB - STUDY OBJECTIVES: To elucidate whether a simple walking aid may improve physical performance in COPD patients with chronic respiratory insufficiency who usually carry their own heavy oxygen canister. DESIGN: Randomized crossover trial. SETTING: Physiopathology laboratory of three rehabilitation centers. PATIENTS AND INTERVENTIONS: We studied 60 stable COPD patients (mean age, 70.6 +/- 7.9 years; FEV(1), 44.8 +/- 14.3% of predicted [+/- SD]) with chronic respiratory insufficiency who randomly performed, on 2 consecutive days, a standardized 6-min walking test using two different modalities: a full-weight oxygen canister transported using a small wheeled cart and pulled by the patient (Aid modality) or full-weight oxygen canister carried on the patient's shoulder (No-Aid modality). MEASUREMENTS AND RESULTS: The distance walked, peak effort dyspnea, and leg fatigue scores as primary outcomes, and other cardiorespiratory parameters as secondary outcomes were recorded during both tests. A significant difference (p < 0.05) between the two tests occurred for all the measured outcomes in favor of the Aid modality. Most importantly, significant changes for distance (+ 43 m, p < 0.001), peak effort dyspnea (- 2.0 points, p < 0.001), leg fatigue (- 1.4 points, p < 0.001), as well as for mean and nadir oxygen saturation and heart rate with the Aid modality (but not with the No-Aid modality) were recorded in the subgroup of patients walking < 300 m at baseline. CONCLUSIONS: This study suggests that a simple walking aid may be helpful in COPD patients receiving long-term oxygen therapy, particularly in those with lower residual exercise capacity. PMID- 17426212 TI - Normal bronchial blood flow in COPD is unaffected by inhaled corticosteroids and correlates with exhaled nitric oxide. AB - BACKGROUND: In COPD patients, there is reduced vascularity and inflammation of the bronchi, which may have opposite effects on bronchial blood flow (QAW). We studied the relationship of QAW with the fraction of exhaled nitric oxide (FENO), which is a potent vasodilator. We also investigated the vascular response to budesonide and a beta(2)-agonist. METHODS: We measured QAW in 17 patients with COPD (mean [+/- SEM] age, 67 +/- 3 years; 10 male patients; mean FEV(1), 57 +/- 3% predicted; mean FEV(1)/FVC ratio, 54 +/- 4%), all of whom were ex-smokers, and in 16 age-matched nonsmoking volunteers (mean age, 64 +/- 4 years) and compared this to FENO. QAW was measured using the acetylene dilution method. RESULTS: Mean QAW was similar in patients with COPD (34.29 +/- 1.09 microL/mL/min) compared to healthy subjects (35.50 +/- 1.74 microL/mL/min; p > 0.05) and was not affected by long-term treatment (35.89 +/- 1.63 microL/mL/min) or short-term treatment (32.50 +/- 1.24 microL/mL/min; p < 0.05) with inhaled budesonide. QAW positively correlated with the diffusion of carbon monoxide (ie, carbon monoxide transfer coefficient: r = 0.74; p < 0.05). FENO levels were mildly elevated in steroid treated patients (10.89 +/- 0.87 parts per billion [ppb]) and untreated patients (9.40 +/- 0.86 ppb) compared to the control group (8.22 +/- 0.57 ppb; p < 0.05) and were correlated with QAW (r = 0.6; p < 0.05). Ten minutes after the inhalation of 200 microg of albuterol, QAW was more elevated in healthy control subjects (59.33 +/- 2.40 microL/mL/min) compared to COPD patients (38.00 +/- 0.58 microL/mL/min; p < 0.05), indicating that COPD patients may have a reduced bronchial vascular reactivity. CONCLUSIONS: QAW is normal in COPD patients and is not affected by therapy with inhaled corticosteroids or beta(2)-agonists. In addition, QAW correlates with levels of FENO, which may have a regulatory role. PMID- 17426213 TI - Depressed myocardial contractile reserve in patients with obstructive sleep apnea assessed by tissue Doppler imaging with dobutamine stress echocardiography. AB - BACKGROUND: Hypoxia has been suggested to affect myocardial contractile function in patients with obstructive sleep apnea (OSA). We sought to determine whether myocardial contractile reserve (MCR), as evaluated by echocardiographic tissue Doppler imaging with dobutamine stress (TDDS), might be depressed in OSA patients. METHODS: Thirty patients with suspected OSA (25 men and 5 women; mean age, 51 +/- 11 years [+/- SD]) underwent overnight polysomnography and TDDS. Peak myocardial systolic velocity (Sm) and peak myocardial early diastolic velocity (Em) in the 12 myocardial segments of the left ventricular (LV) walls were averaged, and the mean Sm and Em during TDDS were compared between patients with apnea-hypopnea index (AHI) <15/h (group 1, n = 13) and those with AHI >/= 15/h (group 2, n = 17). MCR was calculated as the difference between the resting and peak Sm during TDDS. RESULTS: In both groups, Sm increased dose dependently during TDDS. However, the relative increase in Sm was significantly lower in group 2, resulting in a lower value of MCR (5.5 +/- 1.2 cm/s vs 7.4 +/- 1.3 cm/s, p < 0.001). The Em was lower in group 2 compared with group 1 throughout TDDS. MCR was correlated significantly with AHI (r = - 0.67, p < 0.0001), resting Em (r = 0.53, p < 0.005), and body mass index (r = - 0.46, p < 0.05) independent of the LV mass index. CONCLUSIONS: OSA can affect MCR, implying an etiologic contribution from repetitive hypoxic events. TDDS could identify subtle abnormalities of OSA-related cardiac involvement. PMID- 17426214 TI - Respiratory patterns during sleep in obesity-hypoventilation patients treated with nocturnal pressure support: a preliminary report. AB - BACKGROUND: The obesity-hypoventilation syndrome (OHS), commonly defined as a combination of obesity and diurnal hypercapnia, is efficiently treated using nasal positive pressure ventilation (NPPV). The present study aimed to determine whether nocturnal polysomnography allows detection of respiratory disturbances occurring in patients with OHS treated with NPPV that may interfere with the quality of sleep and of ventilatory support, and are not detected by nocturnal pulse oximetry and capnography. METHODS: Twenty OHS patients in stable clinical condition treated by NPPV for at least 3 months with a bilevel pressure support ventilator were studied. All patients underwent single-night polysomnography under NPPV including transcutaneous measurement of Pco(2) (TcPco(2)). Four types of respiratory events were defined and quantified: patient/ventilator desynchronization, periodic breathing (PB), autotriggering, and apnea-hypopneas. RESULTS: Eleven patients (55%) exhibited desynchronization occurring mostly in slow-wave sleep and rapid eye movement sleep and associated with arousals but not inducing significant changes in TcPco(2) or oxygen saturation using pulse oximetry (Spo(2)). Eight patients (40%) showed a high index of PB, mostly occurring in light sleep and associated with more severe nocturnal hypoxemia. Autotriggering was sporadic and usually limited to one or two breaths, although prolonged and asymptomatic autotriggering occurred in one patient during 10.6% of total sleep time. CONCLUSIONS: Patient/ventilatory asynchrony and PB are respiratory patterns occurring frequently in OHS patients treated using NPPV. Nocturnal monitoring of Spo(2) and TcPco(2), commonly used to assess the efficacy of ventilatory support, do not adequately explore this aspect of therapy that might influence its efficacy as well as sleep quality. PMID- 17426215 TI - Predictors of decreased spontaneous baroreflex sensitivity in obstructive sleep apnea syndrome. AB - BACKGROUND: The impact of obstructive sleep apnea syndrome (OSAS) on the arterial baroreflex, and its significance, is still under debate. We investigated the baroreflex sensitivity (BRS) during sleep in well-selected OSAS patient and control subject cohorts METHODS: We performed a prospective study of 10 non-OSAS subjects, 14 subjects with mild-to-moderate OSAS, and 14 male subjects with severe OSAS subjects. Groups were matched for age, body mass index, and other relevant variables. Subjects had no other disease and were not receiving regular medication. BP was monitored beat-by-beat (Portapres; Finapres Medical Systems; Amsterdam, the Netherlands) at night during polysomnography. Spontaneous BRS was assessed by the sequence technique. Heart-rate correction was also applied to calculate BRS at a heart rate (HR) of 60 beats/min (BRS-60) to account for intersubject variability in baseline HR. Eight suitable patients were treated with continuous positive airway pressure (CPAP), and BRS measurements were repeated 6 weeks later. RESULTS: BRS and BRS-60 were significantly lower in patients with severe OSAS than in patients with mild-to-moderate OSAS and in non OSAS subjects, and a separate sleep-stage analysis revealed this difference to be evident in stage 2 non-rapid eye movement sleep and during nocturnal wakefulness. There was no difference in BRS and BRS-60 between non-OSAS subjects and patients with mild-to-moderate OSAS. In multivariate analysis, the desaturation index was the only independent predictor of depressed BRS. CPAP therapy significantly improved the BRS measures. CONCLUSION: Patients with severe OSAS demonstrate depressed BRS during sleep, which may contribute to the cardiovascular pathophysiology in OSAS patients. PMID- 17426216 TI - Biological lung volume reduction: a new bronchoscopic therapy for advanced emphysema. AB - BACKGROUND: Biological lung volume reduction (BLVR) using biological reagents to remodel and shrink damaged regions of lung has previously been accomplished in sheep with experimental pulmonary emphysema. This report summarizes the initial clinical experience including a 3-month follow-up using this technique in humans. METHODS: An open-label phase 1 trial designed to evaluate the safety of BLVR in patients with advanced heterogeneous emphysema enrolled six patients. Of these, three patients received unilateral treatment at two pulmonary subsegments (group 1) and three patients received unilateral treatment at four pulmonary subsegments (group 2). The incidence of adverse events and changes in pulmonary function test results, symptoms, and exercise capacity were evaluated. RESULTS: The mean (+/- SD) age of the six men enrolled in the study was 66 +/- 5.7 years (age range, 57 to 73 years). BLVR was well tolerated in both treatment groups and was not associated with any serious complications. All patients were discharged from the hospital on posttreatment day 1. Although the primary purpose of the study was to examine safety, improvements were observed in mean vital capacity (+7.2 +/- 9.5%; range, -2% to + 19%), mean residual volume (RV) [-7.8 +/- 8.5%; range, + 1% to 22%], mean RV/total lung capacity ratio (-6.6 +/- 4.7%; range, -1% to -15%), mean 6-min walk distance (+14.5 +/- 18.5%; range, 0 to + 51%), and in mean dyspnea score. On average, group 2 patients experienced greater benefit from BLVR than group 1 patients, suggesting a dose-response pattern. CONCLUSIONS: Preliminary results indicate that BLVR can be safe and may produce benefits in appropriately selected patients with advanced heterogeneous emphysema. PMID- 17426217 TI - Tracheobronchial laceration after balloon dilation for benign strictures: incidence and clinical significance. AB - BACKGROUND: Although balloon dilation is a safe procedure, it can lead to laceration of the airway, causing bleeding, pneumothorax, pneumomediastinum, or mediastinitis. We therefore determined the incidence and clinical significance of tracheobronchial lacerations after balloon dilation for treatment of benign tracheobronchial strictures. METHODS: We evaluated 97 patients who had undergone balloon dilation in 124 sessions for the treatment of benign tracheobronchial strictures. The degree of airway laceration was evaluated bronchoscopically. The cumulative airway patency rate after balloon dilation was compared in patients with and without lacerations using Kaplan-Meier survival curves and log-rank testing. RESULTS: There were 64 tracheobronchial lacerations (51.6%) during the 124 sessions of balloon dilation. Of these, 60 were superficial and 4 were deep, but there were no incidents of transmural laceration. In patients with lacerations, mild chest pain (n = 5), blood-tinged sputum (n = 21), and pneumomediastinum (n = 2) occurred, but all resolved completely within 24 h. All superficial lacerations healed spontaneously within 1 month, and all deep lacerations healed 2 to 9 months after conservative treatment. During the follow up period, the median cumulative airway patency period in patients with and without lacerations was 24 and 4 months (p < 0.05), respectively. CONCLUSION: Laceration secondary to balloon dilation in the tracheobronchial tree is relatively common but rarely progresses to transmural laceration and may improve patency outcomes. PMID- 17426218 TI - Central airway mechanics and flow limitation in acquired tracheobronchomalacia. AB - BACKGROUND: Acquired tracheobronchomalacia (TBM) can cause central airway collapse in patients with COPD and may worsen airflow obstruction and symptoms. It is usually not known whether central airway malacia contributes to airflow obstruction. This study was undertaken to quantify central airway collapsibility and relate it to expiratory flow limitation in patients with TBM. METHODS: Eighty patients evaluated for acquired TBM and 4 healthy control subjects were studied with measurements of central airway narrowing derived from bronchoscopic videotapes and simultaneous pressure measurements in the trachea and esophagus. Tracheal narrowing was assessed by a shape index and plotted against the transtracheal pressure to measure collapsibility. Subsequently, airflow and transpulmonary pressure (PL) were measured to identify expiratory flow limitation during quiet breathing and to determine the critical PL required for maximum expiratory flow. RESULTS: Tracheal collapsibility varied widely among patients. Some had profound tracheal narrowing during quiet breathing, and others showed substantial collapse only during forced exhalation. Of the patients, 15% were not flow limited during quiet breathing, 53% were flow limited throughout exhalation, and 30% were flow limited only during the latter part of the exhalation. Patients with flow limitation at rest showed greater tracheal narrowing than those without (p = 0.009), but the severity of expiratory flow limitation was not closely related to tracheal collapsibility. Twenty-three patients were flow limited during quiet exhalation at PLs that did not cause central airway collapse. CONCLUSIONS: In TBM, central airway collapse is not closely related to airflow obstruction, and expiratory flow limitation at rest often occurs in peripheral airways without central airway collapse. PMID- 17426219 TI - Pleural tuberculosis in the United States: incidence and drug resistance. AB - BACKGROUND: Pleural tuberculosis (TB) should be considered in any patient with a lymphocytic pleural effusion. The diagnostic approach is under debate. Knowledge of pleural TB epidemiology would be beneficial. To help clarify pleural TB epidemiology, we analyzed US national TB surveillance data for 1993 to 2003. METHODS: We compared pleural TB to pulmonary TB (where each was reported as the major site of TB disease, and no additional sites of disease were reported). Applicable statistical tests were performed; p < 0.05 was considered to be significant. RESULTS: From 1993 through 2003, 7,549 cases of pleural TB and 156,779 cases of pulmonary TB were reported (in 2003: pleural TB, 536 cases; pulmonary TB, 10,551 cases). The annual proportion of pleural TB was relatively stable (median rate, 3.6%; range, 3.3 to 4.0%) compared to that for pulmonary TB, which steadily decreased (average annual decrease, 0.9%; p < 0.01). Pleural TB occurred significantly more often than pulmonary TB among persons >/= 65 years old (30.4% vs 23.3%, respectively; p < 0.01), and it occurred significantly less often among children < 15 years old (1.8% vs 6.1%, respectively; p < 0.01) and persons 45 to 64 years old (22.9% vs 27.9%, respectively; p < 0.01). Pleural TB patients (63.4%) were born slightly more often in the United States than were pulmonary TB patients (60.9%; p < 0.01). Drug-resistance patterns of pleural TB broadly reflected those of pulmonary TB. However, isolates from pleural TB patients were less often resistant to at least isoniazid (6.0% vs 7.8%, respectively; p < 0.01) and to at least one first-line TB drug (9.9% vs 11.9%, respectively; p < 0.01) compared with pulmonary TB patients. CONCLUSIONS: Knowledge of pleural TB demographic, clinical, and drug-resistance patterns may assist clinicians in making diagnostic and therapeutic decisions. PMID- 17426220 TI - Diagnostic value of interferon-gamma in tuberculous pleurisy: a metaanalysis. AB - BACKGROUND: Conventional tests are not always helpful in making a diagnosis of tuberculous pleurisy. Many studies have investigated the usefulness of interferon (IFN)-gamma measurements in pleural fluid for the early diagnosis of tuberculous pleurisy. We conducted a metaanalysis to determine the accuracy of IFN-gamma measurements in the diagnosis of tuberculous pleurisy. METHODS: After a systematic review of English-language studies, sensitivity, specificity, and other measures of accuracy of IFN-gamma concentrations in the diagnosis of pleural effusion were pooled using random-effects models. Summary receiver operating characteristic curves were used to summarize overall test performance. RESULTS: Twenty-two studies met our inclusion criteria. The summary estimates for IFN-gamma in the diagnosis of tuberculous pleurisy in the studies included were as follows: sensitivity, 0.89 (95% confidence interval [CI], 0.87 to 0.91); specificity, 0.97 (95% CI, 0.96 to 0.98); positive likelihood ratio, 23.45 (95% CI, 17.31 to 31.78); negative likelihood ratio, 0.11 (95% CI, 0.07 to 0.16); and diagnostic odds ratio, 272.7 (95% CI, 147.5 to 504.2). CONCLUSIONS: IFN-gamma determination is a sensitive and specific test for the diagnosis of tuberculous pleurisy. The measurement of IFN-gamma levels in pleural effusions is thus likely to be a useful tool for diagnosing tuberculous pleurisy. The results of IFN-gamma assays should be interpreted in parallel with clinical findings and the results of conventional tests. PMID- 17426221 TI - Chronic and acute effects of smoking on left and right ventricular relaxation in young healthy smokers. AB - BACKGROUND: Left ventricular (LV) diastolic dysfunction has been observed in cigarette smokers with coronary artery disease. The aim of the study was to assess LV and right ventricular (RV) diastolic function in healthy, young, and slim smokers before and after smoking one cigarette. MATERIAL AND METHODS: The participants were 66 healthy volunteers (age < 40 years; body mass index < 25 kg/m(2)): 33 smokers (study group [HS]) and 33 nonsmokers (control group). Echocardiographic examination was done in the HS before smoking one cigarette (HS 1) and after smoking one cigarette (HS-2). To assess diastolic function of LV and RV mitral valve flow (MVF), pulmonary venous flow (PVF) and tricuspid valve flow (TVF) were evaluated. RESULTS: MVF early to late phase ratio (E/A) was significantly lower in HS-1 and HS-2 than in the control group. The PVF systolic to diastolic phase ratio (S/D) was significantly higher in HS-1 and HS-2 than in the control group. These changes suggest LV diastolic function impairment in the HS, but the MVF pattern remained within the normal range. PVF S/D showed systolic dominance (S/D > 1) typical for impaired LV relaxation and abnormal for this age group. TVF E/A was significantly lower in HS-2 than in HS-1 and control subjects and suggests RV diastolic dysfunction. CONCLUSIONS: The following conclusion are made: (1) MVF and PVF demonstrate LV relaxation impairment in healthy smokers before and after smoking one cigarette; (2) the assessment of PVF is a good method reflecting LV diastolic function changes, even when MVF remains normal; and (3) TVF shows RV relaxation impairment after smoking one cigarette in healthy smokers. PMID- 17426222 TI - Niflumic acid and AG-1478 reduce cigarette smoke-induced mucin synthesis: the role of hCLCA1. AB - BACKGROUND: Cigarette smoke induces bronchial mucus secretion. However, the mechanism of this induction is still unidentified. In this study, we investigated the role of the putative calcium-activated chloride channel 1 (CLCA1) and its blocker, niflumic acid, in cigarette smoke-induced mucin synthesis both in vivo and in vitro. METHODS AND RESULTS: Sprague-Dawley rats were exposed to cigarette smoke for 4 weeks. The CLCA1, epidermal growth factor receptor (EGFR), and MUC5AC expressions were increased in the trachea and lung tissues. Goblet-cell hyperplasia with marked mucin staining was detected in the tracheal and bronchial epithelium. In the human bronchial epithelial cell line NCI-H292, cigarette smoke solution also induced mucin production as well as the RNA and protein expressions of CLCA1, EGFR, and MUC5AC. Both in vivo and in vitro, the induction of MUC5AC and mucin synthesis were inhibited by niflumic acid, and/or a selective EGFR tyrosine kinase inhibitor, AG-1478. Niflumic acid also blocked the epidermal growth factor-induced MUC5AC and mucin staining in the NCI-H292 cell line. CONCLUSION: Both EGFR and niflumic acid-sensitive chloride channels (probably CLCA1) are dependently affecting the mucin production as a part of a single complex signaling pathway. CLCA1 may be a key signaling member that can be targeted with pharmacologic interventions to treat mucus hypersecretion. PMID- 17426223 TI - Effectiveness of an elderly smoking cessation counseling training program for social workers: a longitudinal study. AB - BACKGROUND: To achieve greater coverage of the elderly smoking population, the provider/client interface could be broadened to include other professional groups who work with the elderly. We evaluated the effectiveness of a 9-h smoking cessation counseling training program for social workers. METHODS: We recruited 177 social workers and used a preintervention/postintervention longitudinal design, analyzed by multilevel, multivariable modeling to adjust for between subjects covariables and within-subjects correlation in repeated measurements at baseline, 3 months, 6 months, and 12 months after training. RESULTS: Overall, knowledge improved from a mean score of 6.70 +/- 1.03 (+/- SD) at baseline to 7.35 +/- 0.75 at 12 months (range, 0 to 8 correct responses), attitude from 2.84 +/- 0.41 to 3.10 +/- 0.48, and self-perceived competence from 2.49 +/- 0.38 to 2.85 +/- 0.36 (range, 1 to 4, where 4 is best). On multilevel modeling, three of the four "A"s (ask, advice, assist, arrange as per the Agency for Healthcare Research and Quality framework) registered significant gains from baseline to 12 months overall, whereas "advice" did not show any appreciable change. CONCLUSION: These findings demonstrate that our smoking cessation training program achieved sustained effectiveness in the first year after training in enhancing knowledge, positively shifting attitudes, boosting self-perceived competence, and increasing the self-reported frequency of practicing three of the four As in their routine interaction with elderly clients. PMID- 17426224 TI - Gastroesophageal reflux disease, acid suppression, and Mycobacterium avium complex pulmonary disease. AB - BACKGROUND: Weekly symptoms of gastroesophageal reflux disease (GERD) occur in 20% of the population, and GERD has been implicated in the pathophysiology of many respiratory diseases. Microaspiration of contaminated water is a potential portal of entry for Mycobacterium avium complex (MAC) organisms into the respiratory tract, and acid-suppression therapy may enhance the survival of mycobacteria in the stomach. This study aimed to assess the prevalence of GERD, swallowing disorders, reflux symptoms, and acid-suppression therapy in patients with MAC lung disease (MAC positive [MAC+]), and to compare these patients to control subjects without MAC lung disease (MAC negative [MAC-]). METHODS: Clinical information was collected on 58 MAC+ patients and 58 age- and sex matched MAC- patients who were asked to complete a DeMeester questionnaire of reflux symptoms and to identify any acid-suppressive medication consumed. RESULTS: A clinical diagnosis of GERD was documented in 23 of 52 MAC+ patients (44.2%), compared to 16 MAC- patients (27.6%) [p = 0.019]. MAC+ patients consumed significantly more histamine type 2 receptor antagonists and prokinetic agents, and MAC- patients consumed more antacids. The mean DeMeester questionnaire score (+/- SD) for MAC+ patients was 1.39 +/- 1.8, and for MAC- patients was 0.88 +/- 1.4. (p = 0.098). Aspiration was suspected in nine MAC+ patients (15.5%), compared to three MAC- patients (5.2%) [p = 0.032]. There was no association between GERD and radiologic presentation of MAC disease. Consolidation and nodules > 5 mm were more common in those receiving acid suppression than those who were not. CONCLUSIONS: GERD, acid suppression, and clinically suspected aspiration are more common in patients with MAC lung disease than in similar patients without MAC disease. PMID- 17426225 TI - Serum indicators for the diagnosis of pneumocystis pneumonia. AB - BACKGROUND: The diagnosis of pneumocystis pneumonia (PCP) is difficult because it requires microscopic examination to identify pneumocystis from induced sputum or BAL fluid. STUDY OBJECTIVE: To evaluate the usefulness of four serum markers lactate dehydrogenase (LDH), (1-->3) beta-D-glucan (beta-D-glucan), KL-6, and C reactive protein (CRP)-in the diagnosis of PCP. DESIGN: Case-control retrospective study. PATIENTS AND MEASUREMENTS: We reviewed the medical records of 295 consecutive patients who underwent BAL for the diagnosis of PCP. Differential cell counts in BAL fluid and serum levels of LDH, beta-D-glucan, KL 6, and CRP were examined. Oxygenation index was determined using arterial oxygen tension and inspiratory oxygen concentration. RESULTS: Based on the microscopic examination of BAL fluid, 57 patients were PCP positive and 238 patients were PCP negative. There were no significant differences in cell count or differentials in BAL fluid between the positive and negative cases. Serum levels of LDH, beta-D glucan, and KL-6 were significantly higher in PCP-positive patients (p < 0.01). Receiver operating characteristic curves suggest that beta-D-glucan was the most reliable indicator. The cut-off level of beta-D-glucan was estimated to be 31.1 pg/mL, with which the positive and negative predictive values were 0.610 and 0.980, respectively. In PCP-positive patients, the oxygenation index was decreased and correlated with LDH. Both LDH and beta-D-glucan levels were correlated with the proportion of neutrophils in BAL fluid. CONCLUSIONS: Serum beta-D-glucan is a reliable marker for the diagnosis of PCP. Since BAL procedure is invasive, measuring beta-D-glucan should be considered as a primary modality for a diagnosis of PCP, especially for patients with severe respiratory failure. PMID- 17426226 TI - Effect of albuterol on maximal exercise capacity in cystic fibrosis. AB - BACKGROUND: Inhaled, short-acting beta-adrenergic agonists (SAbetaAs) are widely prescribed in cystic fibrosis (CF) subjects, despite a lack of convincing data for efficacy and the potential for these agents to result in airway instability. We tested the hypothesis that inhaled albuterol would improve maximal exercise performance in CF subjects with airflow obstruction, as a result of acute bronchodilation. METHODS: Randomized, double-blind, placebo-controlled crossover study of the effect of inhaled albuterol on maximal exercise performance in 20 stable adult CF patients (mean +/- SD age, 23.3 +/- 6.1 years; FEV(1), 57.65 +/- 17.13% of predicted). RESULTS: Ventilatory limitation to exercise was demonstrated in 16 subjects (80%). Significant bronchodilation occurred with exercise alone (end-exercise FEV(1), 2.24 +/- 0.8 L; vs preexercise FEV(1), 2.09 +/- 0.77 L; p < 0.0001), but albuterol resulted in significantly greater exercise induced bronchodilation than placebo (change in FEV(1), 0.3 +/- 0.15 L vs 0.15 +/ 0.11 L; 95% confidence interval [CI], + 0.07 to + 0.23; p < 0.001). However, there was no difference in maximal workload achieved (albuterol, 158 +/- 46 W; vs placebo, 158 +/- 45 W; 95% CI, - 4.41 to + 4.71; p = 0.95), nor any other measure of exercise performance including maximal oxygen uptake. CONCLUSIONS: Despite causing significant acute bronchodilation, inhaled albuterol did not improve maximal exercise performance in ventilatory-limited CF adults, adding to the body of literature that fails to show any clinical benefit of SAbetaAs in CF subjects. The current results provide further evidence to question the widespread use of these agents, although the potential for adrenergic beta-agonists to instead improve submaximal exercise performance merits further investigation. PMID- 17426227 TI - Infection with Burkholderia cepacia complex bacteria and pulmonary exacerbations of cystic fibrosis. AB - BACKGROUND: Studies have shown that cystic fibrosis (CF) patients who are chronically infected with Burkholderia cepacia complex bacteria may potentially acquire new strains of B cepacia. Our objective was to determine whether pulmonary exacerbations of CF are associated with acquisition of new B cepacia strains or with B cepacia strain replacement. METHODS: Thirty-six patients from seven centers who were chronically infected with B cepacia complex bacteria were prospectively followed up over a 38-month period. Patients had sputum cultures performed every 3 months while clinically stable and at the time of a pulmonary exacerbation. Each B cepacia complex isolate was speciated by polymerase chain reaction amplification of the recA gene to determine species status and was genotyped by pulsed-field gel electrophoresis to determine strain type. RESULTS: Thirty-five of 36 patients (97%) had chronic infection with Burkholderia cenocepacia III-A during clinical stability. All 36 patients maintained the same species and strain of B cepacia complex at the time of exacerbation as was found during clinical stability. B cepacia complex isolates retrieved during exacerbations were significantly less susceptible to ciprofloxacin, chloramphenicol, piperacillin, meropenem, and tobramycin compared to isolates retrieved from the same patients during clinical stability. CONCLUSION: Adult CF patients infected with B cenocepacia maintain the same strain of B cenocepacia during exacerbations; pulmonary exacerbations are not caused by acquisition of a new B cepacia species or strain. B cepacia isolates retrieved during exacerbations may be more resistant to antibiotics. PMID- 17426228 TI - Excess respiratory symptoms in full-time male and female workers in large-scale swine operations. AB - BACKGROUND: The respiratory health effects of working in swine operations have been previously investigated mainly in male owner/operators with intermittent exposure to indoor air contaminants. OBJECTIVES: To examine the respiratory health of male and female workers employed full time in large-scale intensive swine operations in Saskatchewan. DESIGN: A cross-sectional study of male and female swine workers employed full time and nonfarming control subjects. RESULTS: In total, 374 swine farmers (240 men and 134 women) and 411 nonfarming rural control subjects (184 men and 227 women) participated in the study. After controlling for age and smoking, male and female workers were significantly more likely to have chronic and usual cough, and chronic and usual phlegm in comparison to male and female nonfarming control subjects, respectively. The risks of these symptoms were greater in female workers (chronic cough: odds ratio [OR], 5.14; 95% confidence interval, 2.67 to 9.89; chronic phlegm: OR, 4.26; 95% confidence interval, 1.86 to 9.73) than in male workers (chronic cough: OR, 3.47; 95% confidence interval, 1.77 to 6.81; chronic phlegm: OR, 3.22; 95% confidence interval, 1.76 to 5.89). These increased risks were not observed for asthma and asthma-like symptoms in female workers. Male workers had an increased risk of shortness of breath in comparison to their nonfarming counter parts. CONCLUSIONS: Swine workers had increased risk of chronic and usual bronchitis-like symptoms. Female workers in swine operations appear to have greater risk of these symptoms. Future surveillance programs should include both male and female workers. PMID- 17426229 TI - Recent advances in community-acquired pneumonia: inpatient and outpatient. AB - Community-acquired pneumonia (CAP) is a common illness, with the majority of patients treated out of the hospital, yet the greatest burden of the cost of care comes from inpatient management. In the past several years, the management of these patients has advanced, with new information about the natural history and prognosis of illness, the utility of serum markers to guide management, the use of appropriate clinical tools to guide the site-of-care decision, and the finding that guidelines can be developed in a way that improves patient outcome. The challenges to patient management include the emergence of new pathogens and the progression of antibiotic resistance in some of the common pathogens such as Streptococcus pneumoniae. Few new antimicrobial treatment options are available, and the utility of some new therapies has been limited by drug-related toxicity. Ancillary care for severe pneumonia with activated protein C and corticosteroids is being studied, but recently, inpatient care has been most affected by the development of evidence-based "core measures" for management that have been promoted by the Centers for Medicare and Medicaid Services, which form the basis for the public reporting of hospital performance in CAP care. PMID- 17426230 TI - Single-nucleotide polymorphisms and lung disease: clinical implications. AB - Human genetic variation has enormous implications for individual susceptibility to lung disease, as well as for differences in prognosis and response to therapeutic interventions. Single-nucleotide polymorphisms (SNPs) are the most common type of DNA sequence variation. An SNP is the substitution of a single base in the sequence for one that is different from that present in the majority of the population. In this review, we describe in more detail what SNPs are, how they are discovered, and their potential to elucidate the genetic basis of lung disease. We illustrate several examples of how SNPs are being used-or are poised for use-in diagnostic and therapeutic applications. We conclude with a brief discussion of the future of medicine and how genetic knowledge and application can play an ever-increasing and important role in more effective diagnosis and treatment at a more personalized level. PMID- 17426231 TI - Special considerations for patients with cystic fibrosis undergoing lung transplantation. AB - This article reviews lung transplantation in patients with cystic fibrosis (CF). Lung transplantation is commonly utilized for patients with end-stage CF. There are several characteristics of CF that present unique challenges before and after lung transplantation. There is new information available that can be utilized to predict outcomes in patients with end-stage CF, and therefore can help in decisions of referral and listing for lung transplantation. The new lung allocation score, which allocates organs to patients who are on the lung transplant waiting list in the United States, presents new challenges and opportunities for patients with end-stage CF. In addition, the effect of the presence of microbiological flora prior to lung transplantation has been better linked to outcomes after lung transplantation. It is now known that, other than those patients harboring Burkholderia cepacia in their lungs before transplantation, most CF patients can undergo transplantation successfully. Nutrition remains an important issue among CF patients, and diabetes is a common problem after lung transplantation. In contrast, liver disease does not usually present major problems but, if it is severe, can necessitate liver and lung transplantation. Mechanical ventilation prior to transplantation might not be an absolute contraindication for CF patients. CF lung transplant recipients have good outcomes after lung transplantation compared with those of other lung transplant recipients. Quality of life is dramatically improved. However, they are still prone to common complications that all lung transplant recipients are prone to, including primary graft dysfunction, acute and chronic rejection, a variety of infections and malignancies, and renal failure. PMID- 17426232 TI - Catheter-tract metastases associated with chronic indwelling pleural catheters. AB - Indwelling pleural catheters are increasingly being used for ambulatory treatment of malignant pleural effusion, particularly for patients unsuitable for pleurodesis. These catheters are often left in situ for the rest of the patient's life. Tumor metastasis along the tract between pleura and skin surface is a potential complication in patients with chronic indwelling pleural catheters that has seldom been reported. We describe four cases of catheter-tract metastasis that developed between 3 weeks and 9 months after catheter insertion. Catheter tract metastasis occurred in two patients with mesothelioma despite prophylactic irradiation at time of insertion, and in two patients with metastatic adenocarcinoma. All cases were successfully treated using external-beam radiotherapy without necessitating catheter removal. A retrospective audit in our center showed that catheter-tract metastasis occurred in 6.7% of 45 patients treated with indwelling pleural catheters for malignant pleural effusions. Both clinicians and patients should be aware of this potential complication. PMID- 17426233 TI - Flavored cigar smoking induces acute eosinophilic pneumonia. AB - Two cases of acute eosinophilic pneumonia (AEP) following smoking of flavored cigars were analyzed for characteristic features. None of our patients had a history of smoking flavored cigars/cigarettes in the past. One of them had never smoked, and the second patient was an ex-smoker who quit 17 years ago. Both patients presented with community-acquired pneumonia-like symptoms that did not respond to treatment with antibiotics. Their chest radiographs revealed bilateral diffuse infiltrates. The diagnosis of AEP was established based on the clinical picture, BAL that revealed an average eosinophil count > 45%, and immediate clinical improvement after introducing corticosteroids. All other possible causes were excluded during the initial workup. PMID- 17426234 TI - Corynebacterium ulcerans infection of the lung mimicking the histology of Churg Strauss syndrome. AB - We report the first case of pulmonary Corynebacterium ulcerans infection mimicking Churg-Strauss syndrome (CSS). Productive cough, fever, general fatigue, and weight loss developed in a 50-year-old man. Laboratory data revealed prominent eosinophilia and elevated serum IgE. On chest images, multiple nodules and cavities were predominantly detected in the right lung. Histopathologic examination showed necrotizing granulomas and vasculitis with massive eosinophilic infiltration identical to the findings seen in CSS; however, clusters of Gram-positive, coryneform rods were observed in the alveolar spaces. A toxigenic strain of C ulcerans was isolated from lung tissue. The patient was treated with antibiotics, and a favorable clinical course ensued. PMID- 17426235 TI - Adenocarcinoma of the lung presenting as a mycetoma with an air crescent sign. AB - An 89-year-old man was admitted to the hospital due to intermittent anterior chest wall pain for > 1 month. A chest radiograph obtained on November 9, 2004, demonstrated a mass with an irregular border, inside a thin-walled cavity, located in the superior segment of the left lower lobe. A chest CT scan revealed an irregular thin-walled cavity, 5.9 x 5.4 x 4 cm in size, with an air-crescent sign in the superior segment of the left lower lobe, and an intracavitary fungus ball-like mass. A bronchoscopic examination was performed, revealing only external compression of the left lower lobe bronchial lumen. Cultures from both the brushing cytology and brushing fungus specimens were negative. Since the patient was a heavy smoker and the chest radiograph obtained 23 months before had revealed no active pulmonary lesion, neoplastic growth was still highly suspected. Thus, an (18)F-fluoro-2-deoxyglucose positron emission tomography study was performed on November 25, and a mass with a slightly increased standard uptake value (3.17; cutoff value, 2.5) was found. He received a left lower lobe lobectomy on December 23, and a tumor with many septum-like structures connecting the surrounding pulmonary parenchymal tissue was found in the superior segment of the left lower lobe. The final pathologic diagnosis was adenocarcinoma of the lung (pT2N0M0). Thus, even though the chest radiograph and chest CT scan showed a typical air-crescent sign (ie, mass inside a cavity) favoring a mycetoma, the physician should still keep in mind that lung cancer may also unusually present in this way. PMID- 17426236 TI - Hemoglobin Bassett produces low pulse oximeter and co-oximeter readings. AB - Variant hemoglobins can have altered oxygen affinity and can produce changes in oximeter readings. We present a case of hemoglobin Bassett, a possible cause of low pulse oximeter and co-oximeter readings in a 63-year-old woman. PMID- 17426237 TI - A dying art?: The doctor's letter of condolence. PMID- 17426238 TI - A 56-year-old man with choking, recurrent pneumonia, and weight loss. PMID- 17426239 TI - A 45-year-old woman with systemic lupus erythematosus and progressive dyspnea. PMID- 17426240 TI - A 45-year-old man with severe dysphagia and pulmonary infiltrates. PMID- 17426241 TI - Clinical identification of the simple sleep-related movement disorders. AB - Simple sleep-related movement disorders must be distinguished from daytime movement disorders that persist during sleep, sleep-related epilepsy, and parasomnias, which are generally characterized by activity that appears to be simultaneously complex, goal-directed, and purposeful but is outside the conscious awareness of the patient and, therefore, inappropriate. Once it is determined that the patient has a simple sleep-related movement disorder, the part of the body affected by the movement and the age of the patient give clues as to which sleep-related movement disorder is present. In some cases, all-night polysomnography with accompanying video may be necessary to make the diagnosis. Hypnic jerks (ie, sleep starts), bruxism, rhythmic movement disorder (ie, head banging/body rocking), and nocturnal leg cramps are discussed in addition to less well-appreciated disorders such as benign sleep myoclonus of infancy, excessive fragmentary myoclonus, and hypnagogic foot tremor/alternating leg muscle activation. PMID- 17426242 TI - Severe sepsis and septic shock: should blood be transfused to raise mixed venous oxygen saturation? PMID- 17426243 TI - Stress-induced pulmonary systolic hypertension in patients with scleroderma. PMID- 17426244 TI - Partnership with patients: how much partnership? PMID- 17426245 TI - Survey on quality of training in pulmonary physiology during fellowship. PMID- 17426246 TI - Risks in using nonrigorous spanish translations of asthma questionnaires. PMID- 17426247 TI - Inhibitor of DNA binding 1 activates vascular endothelial growth factor through enhancing the stability and activity of hypoxia-inducible factor-1alpha. AB - Inhibitor of DNA binding 1 (Id-1) has been implicated in tumor angiogenesis by regulating the expression of vascular endothelial growth factor (VEGF), but its molecular mechanism has not been fully understood. Here, we show the cross talk between Id-1 and hypoxia-inducible factor-1alpha (HIF-1alpha), that Id-1 induces VEGF by enhancing the stability and activity of HIF-1alpha in human endothelial and breast cancer cells. Although both the transcript and proteins levels of VEGF were induced by Id-1, only the protein expression of HIF-1alpha was induced without transcriptional changes in both human umbilical endothelial cells and MCF7 breast cancer cells. Such induction of the HIF-1alpha protein did not require de novo protein synthesis but was dependent on the active extracellular response kinase (ERK) pathway. In addition, stability of the HIF-1alpha protein was enhanced in part by the reduced association of the HIF-1alpha protein with von Hippel-Lindau protein in the presence of Id-1. Furthermore, Id-1 enhanced nuclear translocation and the transcriptional activity of HIF-1alpha. Transcriptional activation of HIF-1-dependent promoters was dependent on the active ERK pathway, and the association of HIF-1alpha protein with cyclic AMP responsive element binding protein was enhanced by Id-1. Finally, Id-1 induced tube formation in human umbilical endothelial cells, which also required active ERK signaling. In conclusion, we provide the molecular mechanism of the cross talk between HIF-1alpha and Id-1, which may play a critical role in tumor angiogenesis. PMID- 17426248 TI - Integrative genomic analysis of small-cell lung carcinoma reveals correlates of sensitivity to bcl-2 antagonists and uncovers novel chromosomal gains. AB - Cancer is a highly heterogeneous disease in terms of the genetic profile and the response to therapeutics. An early identification of a genomic marker in drug discovery may help select patients that would respond to treatment in clinical trials. Here we suggest coupling compound screening with comparative genomic hybridization analysis of the model systems for early discovery of genomic biomarkers. A Bcl-2 antagonist, ABT-737, has recently been discovered and shown to induce regression of solid tumors, but its activity is limited to a fraction of small-cell lung carcinoma (SCLC) models tested. We used comparative genomic hybridization on high-density single-nucleotide polymorphism genotyping arrays to carry out a genome-wide analysis of 23 SCLC cell lines sensitive and resistant to ABT-737. The screen revealed a number of novel recurrent gene copy number abnormalities, which were also found in an independent data set of 19 SCLC tumors and confirmed by real-time quantitative PCR. A previously unknown amplification was identified on 18q and associated with the sensitivity of SCLC cell lines to ABT-737 and another Bcl-2 antagonist. The region of gain contains Bcl-2 and NOXA, two apoptosis-related genes. Expression microarray profiling showed that the genes residing in the amplified region of 18q are also overexpressed in the sensitive lines relative to the resistant lines. Fluorescence in situ hybridization analysis of tumors revealed that Bcl-2 gain is a frequent event in SCLC. Our findings suggest that 18q21-23 copy number will be a clinically relevant predictor for sensitivity of SCLC to Bcl-2 family inhibitors. The 18q21 23 genomic marker may have a broader application in cancer because Bcl-2 is associated with apoptosis evasion and chemoresistance. PMID- 17426249 TI - Targeted imaging of colonic tumors in smad3-/- mice discriminates cancer and inflammation. AB - The peripheral benzodiazepine receptor (PBR) is a trans-mitochondrial membrane protein that modulates steroid biosynthesis. Recently, up-regulation and nuclear localization of PBR has been shown to be associated with colon, prostate, and breast cancer. PBR has been targeted by the exogenous synthetic ligand, PK11195, for various purposes including imaging. To capitalize on these observations, we developed a high-throughput, noninvasive, in vivo imaging approach to detect spontaneously arising colonic tumors in mice using a novel PBR-targeted molecular imaging agent (NIR-conPK11195). NIR-conPK11195 localized and was retained in colonic adenomas and carcinomas in Smad3(-/-) mice but not in non-neoplastic hamartomas or chronically inflamed colonic tissue. Using a fluorescence signal-to noise ratio of > or =4-fold 13 h after injection of the agent, we detected colonic tumors with a sensitivity of 67% and a specificity of 86% in a cohort of 37 Smad3(-/-) mice and control littermates. Furthermore, using oral administration of dextran sulfate to induce colonic inflammation, we showed that the clearance profile of NIR-conPK11195 distinguished transient uptake in inflammatory tissue from longer term retention in tumors. Taken together, these results indicate that NIR-conPK11195 is a promising optical molecular imaging tool to rapidly screen for colonic tumors in mice and to discriminate inflammation from cancer. PMID- 17426250 TI - Loss of p53 expression correlates with metastatic phenotype and transcriptional profile in a new mouse model of head and neck cancer. AB - Squamous cell carcinoma of the head and neck (HNSCC) is the sixth most frequent cancer worldwide. Because HNSCC is largely acquired by environmental carcinogen exposure rather than through germ line mutations, there are no known familial forms of the disease in humans nor are there inbred rodent strains prone to spontaneous head and neck tumors. Transgenic animals with inactivation of tumor suppressor genes commonly mutated in human cases of HNSCC provide attractive models for studying the pathogenesis of head and neck cancer. p53 is the most frequently inactivated tumor suppressor gene in HNSCC. We used a chemical induction protocol in mice heterozygous for the p53 gene to evaluate how p53 inactivation contributed to head and neck carcinogenesis the mouse model. Metastatic squamous cell carcinomas developed in 100% of animals. Histopathologically, the tumors ranged from well to poorly differentiated and showed many molecular features of human HNSCC. Mice carrying only one p53 allele developed tumors with significantly reduced latency compared with wild-type controls (average, 18 versus 22 weeks). Metastatic cancer cells showed complete loss of p53 expression when compared with primary tumors. Transcriptional profiling showed not only distinct genetic differences between primary and metastatic tumors, but also when cancers from heterozygous null and wild-type animals were compared. Our results provide novel insights into the molecular genetics of tumor progression in head and neck cancer. PMID- 17426251 TI - HMGA2 regulates transcription of the Imp2 gene via an intronic regulatory element in cooperation with nuclear factor-kappaB. AB - IMP2 (insulin-like growth factor-II mRNA binding protein 2) is an oncofetal protein that is aberrantly expressed in several types of cancer. We recently identified the Imp2 gene as a target gene of the architectural transcription factor HMGA2 (high mobility group A2) and its tumor-specific truncated form HMGA2Tr. In this study, we investigated the mechanism via which HMGA2 regulates Imp2 gene expression. We show that HMGA2 and HMGA2Tr directly regulate transcription of the Imp2 gene by binding to an AT-rich regulatory region located in the first intron. In reporter experiments, we show that this AT-rich regulatory region mimics the response of the endogenous Imp2 gene to HMGA2 and HMGA2Tr. Furthermore, we show that a consensus nuclear factor-kappaB (NF-kappaB) binding site located immediately adjacent to the AT-rich regulatory region binds NF-kappaB and that NF-kappaB and HMGA2 cooperate to regulate Imp2 gene expression. Finally, we provide evidence that there is a strong and statistically significant correlation between HMGA2 and IMP2 gene expression in human liposarcomas. PMID- 17426252 TI - Androgen-dependent gene expression of prostate-specific antigen is enhanced synergistically by hypoxia in human prostate cancer cells. AB - The androgen receptor (AR) is implicated in prostate cancer growth, progression, and angiogenesis. Hypoxia-inducible factor-1 (HIF-1), which transcriptionally regulates hypoxia-inducible angiogenic factors, is up-regulated in prostate cancers compared with adjacent normal tissues. HIF-1 may be involved in prostate cancer as well as the AR, but the involvement of HIF-1 in prostate cancer angiogenesis and progression has not been fully elucidated. In the present study, we found that in prostate cancer LNCaP cells dihydrotestosterone enhanced the expression of GLUT-1, one of the HIF-1 target genes, and also that hypoxia enhanced the expression of prostate-specific antigen (PSA) that is one of the AR target genes and is involved in tumor invasion. Small interfering RNA that specifically inhibits HIF-1 reduced the expression levels of PSA as well as GLUT 1. Reporter gene analysis showed that dihydrotestosterone activated the HIF-1 mediated gene expression and hypoxia enhanced the AR-induced promoter activity of human PSA gene. Deletion and site-directed mutation of the 5'-flanking region of human PSA gene revealed that the sequence ACGTG between -3951 and -3947 was essential in the response to hypoxia. Furthermore, chromatin immunoprecipitation assay indicated that HIF-1 interacts with the AR on the human PSA gene promoter. These results indicated that in prostate cancers, HIF-1 might cooperate with the AR to activate the expression of several genes related to tumor angiogenesis, invasion, and progression. PMID- 17426253 TI - Growth stimulation of non-small cell lung cancer cell lines by antibody against epidermal growth factor receptor promoting formation of ErbB2/ErbB3 heterodimers. AB - Antibodies are the most rapidly expanding class of human therapeutics, including their use in cancer therapy. Monoclonal antibodies (mAb) against epidermal growth factor (EGF) receptor (EGFR) generated for cancer therapy block the binding of ligand to various EGFR-expressing human cancer cell lines and abolish ligand dependent cell proliferation. In this study, we show that our mAb against EGFRs, designated as B4G7, exhibited a growth-stimulatory effect on various human cancer cell lines including PC-14, a non-small cell lung cancer cell line; although EGF exerted no growth-stimulatory activity toward these cell lines. Tyrosine phosphorylation of EGFRs occurred after treatment of PC-14 cells with B4G7 mAb, and it was completely inhibited by AG1478, a specific inhibitor of EGFR tyrosine kinase. However, this inhibitor did not affect the B4G7-stimulated cell growth, indicating that the growth stimulation by B4G7 mAb seems to be independent of the activation of EGFR tyrosine kinase. Immunoprecipitation with anti-ErbB3 antibody revealed that B4G7, but not EGF, stimulated heterodimerization between ErbB2 and ErbB3. ErbB3 was tyrosine phosphorylated in the presence of B4G7 but not in the presence of EGF. Further, the phosphorylation and B4G7-induced increase in cell growth were inhibited by AG825, a specific inhibitor of ErbB2. These results show that the ErbB2/ErbB3 dimer functions to promote cell growth in B4G7-treated cells. Changes in receptor-receptor interactions between ErbB family members after inhibition of one of its members are of potential importance in optimizing current EGFR family-directed therapies for cancer. PMID- 17426254 TI - MDM2 splice variants predominantly localize to the nucleoplasm mediated by a COOH terminal nuclear localization signal. AB - Of the >40 alternative and aberrant splice variants of MDM2 that have been described to date, the majority has lost both the well-characterized nuclear localization signal (NLS1) and the nuclear export signal (NES) sequence. Because cellular localization of proteins provides insight regarding their potential function, we determined the localization of three different MDM2 splice variants. The splice variants chosen were the common variants MDM2-A and MDM2-B. In addition, MDM2-FB26 was chosen because it is one of the few variants described that contains the complete p53-binding site. All three splice variants predominantly localized to the nucleus. Nuclear localization of MDM2-A and MDM2-B was controlled by a previously uncharacterized nuclear localization signal (NLS2), whereas nucleoplasmic localization of MDM2-FB26 was mediated by NLS1. p53 and full-length MDM2 colocalized with the splice variants in the nucleus. MDM2-A and MDM2-B both contain a COOH-terminal RING finger domain, and interaction with full-length MDM2 through this domain was confirmed. MDM2-FB26 was the only splice variant evaluated that contained a p53-binding domain; however, interaction between MDM2-FB26 and p53 could not be shown. p14(ARF) did not colocalize with the splice variants and was predominantly expressed within the nucleoli. In summary, nuclear localization signals responsible for the nucleoplasmic distribution of MDM2 splice variants have been characterized. Colocalization and interaction of MDM2-A and MDM2-B with full-length MDM2 in the nucleus have important physiologic consequences, for example, deregulation of p53 activity. PMID- 17426255 TI - An IAP retrotransposon in the mouse ADAMTS13 gene creates ADAMTS13 variant proteins that are less effective in cleaving von Willebrand factor multimers. AB - Severe deficiency of ADAMTS13, a von Willebrand factor (VWF)-cleaving metalloprotease, causes thrombotic thrombocytopenic purpura. When analyzed with VWF multimers, but not with an abbreviated VWF peptide (VWF73) as the substrate, the plasma ADAMTS13 activity levels of mouse strains segregated into a high and a low group that differed by approximately 10 fold. Low ADAMTS13 activity was detected in mice containing 2 alleles of intracisternal A-type particle (IAP) retrotransposon sequence in the ADAMTS13 gene. Molecular cloning of mouse ADAMTS13 identified 2 truncated variants (IAP-a and IAP-b) in the low-activity mice. Both of the IAP variants lacked the 2 carboxyl terminus thrombospondin type 1 repeat (TSR) and CUB domains of full-length ADAMTS13. The IAP-b variant also had splicing abnormalities affecting the spacer domain sequence and had miniscule enzymatic activity. Compared with full-length ADAMTS13, the IAP-a variant was approximately one ninth as active in cleaving VWF multimers but was only slightly less active in cleaving VWF73 peptide. Recombinant human ADAMTS13 was also less effective in cleaving VWF multimers than VWF73 when the C-terminal TSR sequence was deleted. In summary, the carboxyl terminus TSR sequence is important for cleaving VWF multimers. Assay results should be interpreted with caution when peptide substrates are used for analysis of variant ADAMTS13 proteins. PMID- 17426256 TI - Regulation of COX-2 mediated signaling by alpha3 type IV noncollagenous domain in tumor angiogenesis. AB - Human alpha3 chain, a noncollagenous domain of type IV collagen [alpha3(IV)NC1], inhibits angiogenesis and tumor growth. These biologic functions are partly attributed to the binding of alpha3(IV)NC1 to alphaVbeta3 and alpha3beta1 integrins. alpha3(IV)NC1 binds alphaVbeta3 integrin, leading to translation inhibition by inhibiting focal adhesion kinase/phosphatidylinositol 3 kinase/Akt/mTOR/4E-BP1 pathways. In the present study, we evaluated the role of alpha3beta1 and alphaVbeta3 integrins in tube formation and regulation of cyclooxygenase-2 (COX-2) on alpha3(IV)NC1 stimulation. We found that although both integrins were required for the inhibition of tube formation by alpha3(IV)NC1 in endothelial cells, only alpha3beta1 integrin was sufficient to regulate COX-2 in hypoxic endothelial cells. We show that binding of alpha3(IV)NC1 to alpha3beta1 integrin leads to inhibition of COX-2-mediated pro angiogenic factors, vascular endothelial growth factor, and basic fibroblast growth factor by regulating IkappaBalpha/NFkappaB axis, and is independent of alphaVbeta3 integrin. Furthermore, beta3 integrin-null endothelial cells, when treated with alpha3(IV)NC1, inhibited hypoxia-mediated COX-2 expression, whereas COX-2 inhibition was not observed in alpha3 integrin-null endothelial cells, indicating that regulation of COX-2 by alpha3(IV)NC1 is mediated by integrin alpha3beta1. Our in vitro and in vivo findings demonstrate that alpha3beta1 integrin is critical for alpha3(IV)NC1-mediated inhibition of COX-2-dependent angiogenic signaling and inhibition of tumor progression. PMID- 17426257 TI - Increased risk of pregnancy complications in patients with essential thrombocythemia carrying the JAK2 (617V>F) mutation. AB - Essential thrombocythemia (ET) may occur in women of childbearing age. To investigate the risk of pregnancy complications, we studied 103 pregnancies that occurred in 62 women with ET. The 2-tailed Fisher exact test showed that pregnancy outcome was independent from that of a previous pregnancy. The rate of live birth was 64%, and 51% of pregnancies were uneventful. Maternal complications occurred in 9%, while fetal complications occurred in 40% of pregnancies. The Mantel-Haenszel method showed that fetal loss in women with ET was 3.4-fold higher (95% confidence interval [CI]: 3-3.9; P < .001) than in the general population. Half of the women studied carried the JAK2 (617V>F) mutation, and a multivariate logistic regression model identified this mutation as an independent predictor of pregnancy complications (P = .01). Neither the platelet count nor the leukocyte count was a risk factor. JAK2 (617V>F)-positive patients had an odds ratio of 2.02 (95% CI: 1.1 - 3.8) of developing complications in comparison with JAK2 (617V>F)-negative patients. Aspirin did not prevent complication in JAK2 (617V>F)-positive patients and appeared to worsen outcome in JAK2 (617V>F)-negative patients. A relationship was found between JAK2 (617V>F) and fetal loss (P = .05). This study indicates that patients carrying the JAK2 (617V>F) mutation have higher risk of developing pregnancy complications. PMID- 17426258 TI - Constitutive phosphoinositide 3-kinase/Akt activation represents a favorable prognostic factor in de novo acute myelogenous leukemia patients. AB - The phosphoinositide 3-kinase (PI3K/Akt) pathway is activated in acute myelogenous leukemia (AML) and is promising for targeted inhibition. Ninety-two patients with primary AML were analyzed for PI3K/Akt constitutive activation. Fifty percent of the patients presented with constitutive PI3K activation (PI3K (+)). No difference was observed between PI3K (+) and PI3K (-) groups concerning age, sex, white blood cell count, lactate dehydrogenase (LDH) level, bone marrow blast cells, French-American-British (FAB) classification, cytogenetics, RAS or nucleophosmin (NPM) mutations. Slightly more FLT3-ITD was detected in the PI3K ( ) group (P = .048). The complete remission rate was similar between the 2 groups. With a median follow-up of 26 months, we observed for PI3K (+) and PI3K (-) patients, respectively, 56% and 33% overall survival (P = .001) and 72% and 41% relapse-free survival (P = .001). Constitutive PI3K/Akt activity is a favorable prognosis factor in AML, even after adjustment for FLT3-ITD, and may confer a particular sensitivity to chemotherapy. A better understanding of the downstream effectors of the PI3K/Akt pathway is needed before targeting in AML. PMID- 17426259 TI - The effects of widowhood and vascular risk factors on late-life depression. AB - OBJECTIVE: The purpose of this study was to examine whether the impact of conjugal loss on depression is greater in the presence of vascular risk factors (CVRFs) (stress vulnerability hypothesis), or whether conjugal loss and CVRFs are independent predictors of depression following spousal loss (independent pathways hypothesis). METHOD: The current study is a secondary data analysis of the Changing Lives of Older Couples database, which is a study of older widowed persons. One thousand five hundred thirty-two participants engaged in a baseline interview, and interviews were conducted 6, 18, and 48 months after the death of a spouse. RESULTS: Spousal loss is a significant predictor of depressive symptoms at six months after the death. At 18- and 48-months postloss, CVRFs significantly predict depression onset, however no interactions between the two variables were seen at any of the three follow-up waves. Looking longitudinally from baseline to each follow-up wave, the widows with low CVRFs were at greater risk for elevated depression at six-months postloss than the non-widows with low vascular risk, and all widows were at a greater risk for elevated depression at 18-months postloss. At 48-month follow-up, those with high CVRFs who had not lost a spouse were at significantly greater risk for depression than the non-widows with low CVRFs. CONCLUSIONS: Results do not provide support for the stress-vulnerability hypothesis and suggest that loss and CVRFs are independent predictors of depression, whose effects vary with the passage of time. PMID- 17426260 TI - Cognitive impairment in late-life generalized anxiety disorder. AB - OBJECTIVE: This study sought to characterize cognitive functioning in elderly patients with generalized anxiety disorder (GAD), as compared with normal comparison subjects and patients with major depression. METHODS: The cognitive functioning in GAD (N=19) was assessed with the Mattis Dementia Rating Scale and across specific domains of naming, executive ability, and memory, in comparison with late-life major depressive disorder (MDD; N=68) and versus no psychiatric illness (N=40). RESULTS: In comparison to healthy normal comparison subjects, anxious subjects were impaired on measures of short-term and delayed memory. Depressed subjects also performed worse than normal comparison subjects on delayed memory, as well as in naming. Anxious subjects did not differ significantly from depressed subjects in any measure of cognitive function. CONCLUSION: In this preliminary study, anxious subjects displayed cognitive impairments in short-term memory; while depressed patients compared to normal comparison subjects showed executive dysfunction and more general cognitive impairments not evident in anxious subjects. Studies of neuropsychological function in elderly anxious subjects may be informative in developing treatment interventions that mitigate cognitive dysfunction and illuminate the course of illness and underlying neural pathways. PMID- 17426261 TI - The cover. Calla Lilies. PMID- 17426262 TI - A piece of my mind . Steven's violin. PMID- 17426263 TI - Researchers buoyed by novel HIV drugs: will expand drug arsenal against resistant virus. PMID- 17426264 TI - New clues to HDL's benefits revealed. PMID- 17426265 TI - Study aims to clarify efficacy, safety of eye drug treatments. PMID- 17426266 TI - Disease, drug response linked to loss or gain of big DNA chunks in genome. PMID- 17426267 TI - Breastfeeding benefits in some HIV settings. PMID- 17426268 TI - Sham surgery in clinical trials. PMID- 17426269 TI - Nonoperative treatment for lumbar disk herniation. PMID- 17426270 TI - Performance measures and outcomes for patients hospitalized with heart failure. PMID- 17426271 TI - Performance measures and outcomes for patients hospitalized with heart failure. PMID- 17426272 TI - Folic acid supplementation and cardiovascular diseases. PMID- 17426273 TI - Folic acid supplementation and cardiovascular diseases. PMID- 17426274 TI - Nonvalidation of reported genetic risk factors for acute coronary syndrome in a large-scale replication study. AB - CONTEXT: Given the numerous, yet inconsistent, reports of genetic variants being associated with acute coronary syndromes (ACS), there is a need for comprehensive validation of ACS susceptibility genotypes. OBJECTIVE: To perform an extensive validation of putative genetic risk factors for ACS. DESIGN, SETTING, AND PARTICIPANTS: Through a systematic literature search of articles published before March 10, 2005, we identified genetic variants previously reported as significant susceptibility factors for atherosclerosis or ACS. Restricting our analysis to white patients to reduce confounding from racial admixture, we identified 811 patients who presented from March 2001 through June 2003 with ACS at 2 Kansas City, Mo, university-affiliated hospitals. During 2005-2006, we genotyped the 811 patients along with 650 age- and sex-matched controls for 85 variants in 70 genes and attempted to replicate previously reported associations. We further explored possible associations without prior assumption of specific risk models and used the Sign test to search for weak associations. MAIN OUTCOME MEASURES: Compare each prespecified gene variant associated with ACS risk among cases and controls. A surplus of associations would imply that some are associated with ACS. RESULTS: Of 85 variants tested, only 1 putative risk genotype (-455 promoter variant in beta-fibrinogen) was nominally statistically significant (P = .03). Only 4 additional genes were positive in model-free analysis. Neither number of associations was more frequent than expected by chance, given the number of comparisons. Finally, only 41 of 84 predefined risk variants were even marginally more frequent in cases than in controls (with 1 tie), representing a 48.8% "win rate" (95% confidence interval, 38.1%-59.5%) for the collective risk genotypes (P = .91, Sign test). CONCLUSIONS: Our null results provide no support for the hypothesis that any of the 85 genetic variants tested is a susceptibility factor for ACS. These results emphasize the need for robust replication of putative genetic risk factors before their introduction into clinical care. PMID- 17426275 TI - Corticosteroids for the prevention of atrial fibrillation after cardiac surgery: a randomized controlled trial. AB - CONTEXT: Atrial fibrillation (AF) is the most common arrhythmia to occur after cardiac surgery. An exaggerated inflammatory response has been proposed to be one etiological factor. OBJECTIVE: To test whether intravenous corticosteroid administration after cardiac surgery prevents AF after cardiac surgery. DESIGN, SETTING, AND PATIENTS: A double-blind, randomized multicenter trial (study enrollment August 2005-June 2006) in 3 university hospitals in Finland of 241 consecutive patients without prior AF or flutter and scheduled to undergo first on-pump coronary artery bypass graft (CABG) surgery, aortic valve replacement, or combined CABG surgery and aortic valve replacement. INTERVENTION: Patients were randomized to receive either 100-mg hydrocortisone or matching placebo as follows: the first dose in the evening of the operative day, then 1 dose every 8 hours during the next 3 days. In addition, all patients received oral metoprolol (50-150 mg/d) titrated to heart rate. MAIN OUTCOME MEASURE: Occurrence of AF during the first 84 hours after cardiac surgery. RESULTS: The incidence of postoperative AF was significantly lower in the hydrocortisone group (36/120 [30%]) than in the placebo group (58/121 [48%]; adjusted hazard ratio, 0.54; 95% confidence interval, 0.35-0.83; P = .004; number needed to treat, 5.6). Compared with placebo, patients receiving hydrocortisone did not have higher rates of superficial or deep wound infections, or other major complications. CONCLUSION: Intravenous hydrocortisone reduced the incidence of AF after cardiac surgery. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00442494. PMID- 17426276 TI - Autologous nonmyeloablative hematopoietic stem cell transplantation in newly diagnosed type 1 diabetes mellitus. AB - CONTEXT: Type 1 diabetes mellitus (DM) results from a cell-mediated autoimmune attack against pancreatic beta cells. Previous animal and clinical studies suggest that moderate immunosuppression in newly diagnosed type 1 DM can prevent further loss of insulin production and can reduce insulin needs. OBJECTIVE: To determine the safety and metabolic effects of high-dose immunosuppression followed by autologous nonmyeloablative hematopoietic stem cell transplantation (AHST) in newly diagnosed type 1 DM. DESIGN, SETTING, AND PARTICIPANTS: A prospective phase 1/2 study of 15 patients with type 1 DM (aged 14-31 years) diagnosed within the previous 6 weeks by clinical findings and hyperglycemia and confirmed with positive antibodies against glutamic acid decarboxylase. Enrollment was November 2003-July 2006 with observation until February 2007 at the Bone Marrow Transplantation Unit of the School of Medicine of Ribeirao Preto, Ribeirao Preto, Brazil. Patients with previous diabetic ketoacidosis were excluded after the first patient with diabetic ketoacidosis failed to benefit from AHST. Hematopoietic stem cells were mobilized with cyclophosphamide (2.0 g/m2) and granulocyte colony-stimulating factor (10 microg/kg per day) and then collected from peripheral blood by leukapheresis and cryopreserved. The cells were injected intravenously after conditioning with cyclophosphamide (200 mg/kg) and rabbit antithymocyte globulin (4.5 mg/kg). MAIN OUTCOME MEASURES: Morbidity and mortality from transplantation and temporal changes in exogenous insulin requirements (daily dose and duration of usage). Secondary end points: serum levels of hemoglobin A1c, C-peptide levels during the mixed-meal tolerance test, and anti-glutamic acid decarboxylase antibody titers measured before and at different times following AHST. RESULTS: During a 7- to 36-month follow-up (mean 18.8), 14 patients became insulin-free (1 for 35 months, 4 for at least 21 months, 7 for at least 6 months; and 2 with late response were insulin-free for 1 and 5 months, respectively). Among those, 1 patient resumed insulin use 1 year after AHST. At 6 months after AHST, mean total area under the C-peptide response curve was significantly greater than the pretreatment values, and at 12 and 24 months it did not change. Anti-glutamic acid decarboxylase antibody levels decreased after 6 months and stabilized at 12 and 24 months. Serum levels of hemoglobin A(1c) were maintained at less than 7% in 13 of 14 patients. The only acute severe adverse effect was culture-negative bilateral pneumonia in 1 patient and late endocrine dysfunction (hypothyroidism or hypogonadism) in 2 others. There was no mortality. CONCLUSIONS: High-dose immunosuppression and AHST were performed with acceptable toxicity in a small number of patients with newly diagnosed type 1 DM. With AHST, beta cell function was increased in all but 1 patient and induced prolonged insulin independence in the majority of the patients. PMID- 17426277 TI - Safety and immunogenicity of a baculovirus-expressed hemagglutinin influenza vaccine: a randomized controlled trial. AB - CONTEXT: A high priority in vaccine research is the development of influenza vaccines that do not use embryonated eggs as the substrate for vaccine production. OBJECTIVE: To determine the dose-related safety, immunogenicity, and protective efficacy of an experimental trivalent influenza virus hemagglutinin (rHA0) vaccine produced in insect cells using recombinant baculoviruses. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, placebo-controlled clinical trial at 3 US academic medical centers during the 2004-2005 influenza season among 460 healthy adults without high-risk indications for influenza vaccine. INTERVENTIONS: Participants were randomly assigned to receive a single injection of saline placebo (n = 154); 75 microg of an rHA0 vaccine containing 15 microg of hemagglutinin from influenza A/New Caledonia/20/99(H1N1) and influenza B/Jiangsu/10/03 virus and 45 microg of hemagglutinin from influenza A/Wyoming/3/03(H3N2) virus (n = 153); or 135 microg of rHA0 containing 45 microg of hemagglutinin each from all 3 components (n = 153). Serum samples were taken before and 30 days following immunization. MAIN OUTCOME MEASURES: Primary safety end points were the rates and severity of solicited and unsolicited adverse events. Primary immunogenicity end points were the rates of 4-fold or greater increases in serum hemagglutinin inhibition antibody to each of the 3 vaccine strains before and 28 days after inoculation. The prespecified primary efficacy end point was culture-documented influenza illness, defined as development of influenza-like illness associated with influenza virus on a nasopharyngeal swab. RESULTS: Rates of local and systemic adverse effects were low, and the rates of systemic adverse effects were not different in either vaccine group than in the placebo group. Hemagglutinin inhibition antibody responses to the H1 component were seen in 3% of placebo, 51% of 75-microg vaccine, and 67% of 135-microg vaccine recipients, while responses to B were seen in 4% of placebo, 65% of 75 microg vaccine, and 92% of 135-microg vaccine recipients. Responses to the H3 component occurred in 11% of placebo, 81% of 75-microg vaccine, and 77% of 135 microg vaccine recipients. Influenza infections in the study population were due to influenza B and A(H3N2), and influenza A infections were A/California/7/2004 like viruses, an antigenically drifted strain. Seven cases of culture-confirmed CDC-defined influenza-like illness occurred in 153 placebo recipients (4.6%) compared with 2 cases (1.3%) in 150 recipients of 75 microg of vaccine, and 0 cases in recipients of 135 microg of vaccine. CONCLUSIONS: In this study, a trivalent rHA0 vaccine was safe and immunogenic in a healthy adult population. Preliminary evidence of protection against a drifted influenza A(H3N2) virus was obtained, but the sample size was small. Inclusion of a neuraminidase component did not appear to be required for protection. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00328107. PMID- 17426278 TI - Does this patient have ventilator-associated pneumonia? AB - CONTEXT: Ventilator-associated pneumonia (VAP) is a common and serious nosocomial infection. Accurate, timely diagnosis enables affected patients to receive appropriate therapy and avoids mistreatment of patients having other conditions. OBJECTIVE: To review the published medical literature describing the precision and accuracy of clinical, radiographic, and laboratory data to diagnose bacterial VAP relative to a histological gold standard. DATA SOURCES: English-language articles identified by a structured search strategy using MEDLINE (January 1966 October 31, 2006) and Google Scholar. Additional articles were identified through the reference lists of studies and review papers identified by the search strategy. STUDY SELECTION: Included studies described clinical findings associated with VAP in 25 or more patients receiving mechanical ventilation who subsequently underwent pulmonary biopsy or autopsy. Fourteen studies describing clinical findings in 655 patients met inclusion criteria. DATA EXTRACTION: Data were abstracted onto a structured form, allowing calculation of the likelihood ratios (LRs) for each sign or combination of findings. DATA SYNTHESIS: The presence or absence of fever, abnormal white blood cell count, or purulent pulmonary secretions do not substantively alter the probability of VAP. However, the combination of a new radiographic infiltrate with at least 2 of fever, leukocytosis, or purulent sputum increases the likelihood of VAP (summary LR, 2.8; 95% confidence interval, 0.97-7.9). The absence of a new infiltrate on a plain chest radiograph lowers the likelihood of VAP (summary LR, 0.35; 95% confidence interval, 0.14-0.87). Fewer than 50% neutrophils on cell count analysis of lower pulmonary secretions makes VAP unlikely (LR range, 0.05-0.10). CONCLUSIONS: Routine bedside evaluation coupled with radiographic information provides suggestive but not definitive evidence that VAP is present or absent. Given the severity of VAP and the frequency of serious conditions that can mimic VAP, clinicians should be ready to consider additional tests that provide further evidence for VAP or that establish another diagnosis. PMID- 17426279 TI - Back pain in the workplace. PMID- 17426280 TI - Crash risk in general aviation. PMID- 17426281 TI - Cellular therapy for type 1 diabetes: has the time come? PMID- 17426282 TI - JAMA patient page. Ventilator-associated pneumonia. PMID- 17426283 TI - Magnetic resonance imaging parameters for assessing risk of recurrent hamstring injuries in elite athletes. AB - BACKGROUND: Magnetic resonance (MR) imaging has established its usefulness in diagnosing hamstring muscle strain and identifying features correlating with the duration of rehabilitation in athletes; however, data are currently lacking that may predict which imaging parameters may be predictive of a repeat strain. PURPOSE: This study was conducted to identify whether any MR imaging-identifiable parameters are predictive of athletes at risk of sustaining a recurrent hamstring strain in the same playing season. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: Forty-one players of the Australian Football League who sustained a hamstring injury underwent MR examination within 3 days of injury between February and August 2002. The imaging parameters measured were the length of injury, cross-sectional area, the specific muscle involved, and the location of the injury within the muscle-tendon unit. Players who suffered a repeat injury during the same season were reimaged, and baseline and repeat injury measurements were compared. Comparison was also made between this group and those who sustained a single strain. RESULTS: Forty-one players sustained hamstring strains that were positive on MR imaging, with 31 injured once and 10 suffering a second injury. The mean length of hamstring muscle injury for the isolated group was 83.4 mm, compared with 98.7 mm for the reinjury group (P = .35). In the reinjury group, the second strain was also of greater length than the original (mean, 107.5 mm; P = .07). Ninety percent of players sustaining a repeat injury demonstrated an injury length greater than 60 mm, compared with only 58% in the single strain group (P = .01). Only 7% of players (1 of 14) with a strain <60 mm suffered a repeat injury. Of the 27 players sustaining a hamstring strain >60 mm, 33% (9 of 27) suffered a repeat injury. Of all the parameters assessed, only a history of anterior cruciate ligament sprain was a statistically significant predictor for suffering a second strain during the same season of competition. CONCLUSION: A history of anterior cruciate ligament injury was the only statistically significant risk factor for a recurrent hamstring strain in our study. Of the imaging parameters, the MR length of a strain had the strongest correlation association with a repeat hamstring strain and therefore may assist in identifying which athletes are more likely to suffer further reinjury. PMID- 17426284 TI - In vivo imaging of farnesoid X receptor activity reveals the ileum as the primary bile acid signaling tissue. AB - We generated and characterized a firefly luciferase reporter mouse for the nuclear receptor farnesoid X receptor (FXR). This FXR reporter mouse has basal luciferase expression in the terminal ileum, an organ with well-characterized FXRalpha signaling. In vivo luciferase activity reflected the diurnal activity pattern of the mouse, and is regulated by both natural (bile acids, chenodeoxycholic acid) and synthetic (GW4064) FXRalpha ligands. Moreover, in vivo and in vitro analysis showed luciferase activity after GW4064 administration in the liver, kidney, and adrenal gland, indicating that FXRalpha signaling is functional in these tissues. Hepatic luciferase activity was robustly induced in cholestatic mice, showing that FXRalpha signaling pathways are activated in this disease. In conclusion, we have developed an FXR reporter mouse that is useful to monitor FXRalpha signaling in vivo in health and disease. The use of this animal could facilitate the development of new therapeutic compounds that target FXRalpha in a tissue-specific manner. PMID- 17426285 TI - Hes1 and Hes5 control the progenitor pool, intermediate lobe specification, and posterior lobe formation in the pituitary development. AB - The pituitary gland is composed of two distinct entities: the adenohypophysis, including the anterior and intermediate lobes, and the neurohypophysis, known as the posterior lobe. This critical endocrine organ is essential for homeostasis, metabolism, reproduction, and growth. The pituitary development requires the control of proliferation and differentiation of progenitor cells. Although multiple signaling molecules and transcription factors are required for the proper pituitary development, the mechanisms that regulate the fate of progenitor cells remain to be elucidated. Hes genes, known as Notch effectors, play a crucial role in specifying cellular fates during the development of various tissues and organs. Here, we report that mice deficient for Hes1 and Hes5 display severe pituitary hypoplasia caused by accelerated differentiation of progenitor cells. In addition, this hypoplastic pituitary gland (adenohypophysis) lacks the intermediate lobe and exhibits the features of the anterior lobe only. Hes1 and Hes5 double-mutant mice also lack the neurohypophysis (the posterior lobe), probably due to incomplete evagination of the diencephalon. Thus, Hes genes control not only maintenance of progenitor cells but also intermediate vs. anterior lobe specification during the adenohypophysis development. Hes genes are also essential for the formation of the neurohypophysis. PMID- 17426286 TI - Signal transducer and activator of transcription (Stat) 5b-mediated inhibition of insulin-like growth factor binding protein-1 gene transcription: a mechanism for repression of gene expression by growth hormone. AB - GH plays a central role in controlling somatic growth, tissue regeneration, and intermediary metabolism in most vertebrate species through mechanisms dependent on the regulation of gene expression. Recent studies using transcript profiling have identified large cohorts of genes whose expression is induced by GH. Other results have demonstrated that signal transducer and activator of transcription (Stat) 5b, a latent transcription factor activated by the GH receptor-associated protein kinase, Jak2, is a key agent in the GH-stimulated gene activation that leads to somatic growth. By contrast, little is known about the steps through which GH-initiated signaling pathways reduce gene expression. Here we show that Stat5b plays a critical role in the GH-regulated inhibition of IGF binding protein-1 gene transcription by impairing the actions of the FoxO1 transcription factor on the IGF binding protein-1 promoter. Additional observations using transcript profiling in the liver indicate that Stat5b may be a general mediator of GH-initiated gene repression. Our results provide a model for understanding how GH may simultaneously stimulate and inhibit the expression of different cohorts of genes via the same transcription factor, potentially explaining how GH action leads to integrated biological responses in the whole organism. PMID- 17426287 TI - Calcium-sensing receptor endocytosis links extracellular calcium signaling to parathyroid hormone-related peptide secretion via a Rab11a-dependent and AMSH sensitive mechanism. AB - The calcium-sensing receptor (CaR) helps to maintain the homeostasis of extracellular calcium by controlling the secretion of hormones associated with this process. The mechanism of agonist-induced endocytosis and down-regulation of CaR and the influence of this event on the secretion of CaR-regulated hormones is not fully understood. In this study, we show that CaR is constitutively endocytosed and recycled to the plasma membrane by a Rab11a-dependent mechanism; during this process, the level of total cellular CaR is maintained. This trafficking of CaR promotes the secretion of PTH-related peptide (PTHrP), as evidenced by a decrease on PTHrP secretion in the presence of a dominant-negative mutant of Rab11a. Interestingly, this Rab11a dominant-negative mutant does not interfere with CaR-dependent activation of ERK 1/2, suggesting that ERK signaling is not sufficient to promote PTHrP secretion downstream of CaR. In addition, AMSH (associated molecule with the SH3 domain of STAM), a CaR carboxyl-terminal binding protein, redirects CaR from slow recycling to down-regulation, reducing CaR expression and decreasing PTHrP secretion. Our results indicate that endocytosis and trafficking of CaR modulate PTHrP secretion. PMID- 17426288 TI - Surface-scanning mutational analysis of protein arginine methyltransferase 1: roles of specific amino acids in methyltransferase substrate specificity, oligomerization, and coactivator function. AB - Protein arginine methyltransferase 1 (PRMT1) is an arginine-specific protein methyltransferase that methylates a number of proteins involved in transcription and other aspects of RNA metabolism. Its role as a transcriptional coactivator for nuclear receptors involves its ability to bind to other coactivators, such as glucocorticoid receptor-interacting protein 1 (GRIP1), as well as its ability to methylate histone H4 and coactivators such as peroxisome proliferator-activated receptor gamma coactivator-1alpha. Its ability to form homodimers or higher-order homo-oligomers also is important for its methyltransferase activity. To understand the function of PRMT1 further, 19 surface residues were mutated, based on the crystal structure of PRMT1. Mutants were characterized for their ability to bind and methylate various substrates, form homodimers, bind GRIP1, and function as a coactivator for the androgen receptor in cooperation with GRIP1. We identified specific surface residues that are important for methylation substrate specificity and binding of substrates, for dimerization/oligomerization, and for coactivator function. This analysis also revealed functional relationships between the various activities of PRMT1. Mutants that did not dimerize well had poor methyltransferase activity and coactivator function. However, surprisingly, all dimerization mutants exhibited increased GRIP1 binding, suggesting that the essential PRMT1 coactivator function of binding to GRIP1 may require dissociation of PRMT1 dimers or oligomers. Three different mutants with altered substrate specificity had widely varying coactivator activity levels, suggesting that methylation of specific substrates is important for coactivator function. Finally, identification of several mutants that exhibited reduced coactivator function but appeared normal in all other activities tested, and finding one mutant with very little methyltransferase activity but normal coactivator function, suggested that these mutated surface residues may be involved in currently unknown protein-protein interactions that are important for coactivator function. PMID- 17426289 TI - SORCS1: a novel human type 2 diabetes susceptibility gene suggested by the mouse. AB - OBJECTIVE: A small number of susceptibility genes for human type 2 diabetes have been identified by candidate gene analysis or positional cloning. Genes found to influence diabetes or related traits in mice are likely to be susceptibility genes in humans. SorCS1 is the gene identified as responsible for the mouse chromosome 19 T2dm2 quantitative trait locus for fasting insulin levels, acting via impaired insulin secretion and increased islet disruption in obese females. Genes that impair compensatory insulin secretion in response to obesity-induced insulin resistance may be particularly relevant to human diabetes. Thus, we sought to determine whether variation in the human SORCS1 gene was associated with diabetes-related traits. RESEARCH DESIGN AND METHODS: We assessed the contribution of variation in SORCS1 to human insulin-related traits in two distinct Mexican-American cohorts. One cohort (the Mexican-American Coronary Artery Disease [MACAD] cohort) consisted of nondiabetic individuals, allowing assessment of genetic association with subclinical intermediate insulin-related traits; the second cohort (the San Antonio Family Diabetes Study [SAFADS]) contained individuals with diabetes, allowing association analyses with overt disease. RESULTS: We first found association of SORCS1 single nucleotide polymorphisms and haplotypes with fasting insulin levels and insulin secretion in the MACAD cohort. Similar to our results in the mice, the genetic association was strongest in overweight women. We then observed association with diabetes risk and age at diagnosis in women of the SAFADS cohort. CONCLUSIONS: Identification of SORCS1 as a novel gene affecting insulin secretion and diabetes risk is likely to provide important insight into the biology of obesity-induced type 2 diabetes. PMID- 17426290 TI - Surgical exposures of the humerus. PMID- 17426291 TI - Interspinous process spacers. AB - The patient with neurogenic claudication resulting from lumbar spinal stenosis who fails to experience satisfactory relief from nonsurgical measures has limited treatment options. Lumbar epidural steroid injections and surgical laminectomy are generally accepted alternatives for the patient with moderate to severe symptoms. Interspinous process spacers, a relatively new class of technology, are proposed for use in the patient who prefers less invasive surgery or in whom medical comorbidities preclude a major surgical procedure. Early data from biomechanical and clinical studies support the short-term efficacy of interspinous process spacers in treating claudication related to spinal stenosis. Sufficient medium- and long-term data are lacking, however, particularly with respect to durability of symptomatic relief and the risk of device migration or dislocation. Although interspinous process spacers are a promising new technology, the results of longer-term clinical follow-up studies are needed to more clearly define their role in the management of lumbar spinal stenosis. PMID- 17426292 TI - Osteonecrosis in the foot. AB - Osteonecrosis, also referred to as avascular necrosis, refers to the death of cells within bone caused by a lack of circulation. It has been documented in bones throughout the body. In the foot, osteonecrosis is most commonly seen in the talus, the first and second metatarsals, and the navicular. Although uncommon, osteonecrosis has been documented in almost every bone of the foot and therefore should be considered in the differential diagnosis when evaluating both adult and pediatric foot pain. Osteonecrosis is associated with many foot problems, including fractures of the talar neck and navicular as well as Kohler's disease and Freiberg's disease. Orthopaedists who manage foot disorders will at some point likely be faced with the challenges associated with patients with osteonecrosis of the foot. Because this disease can masquerade as many other pathologies, physicians should be aware of the etiology, presentation, and treatment options for osteonecrosis in the foot. PMID- 17426293 TI - The rotator interval: anatomy, pathology, and strategies for treatment. AB - Over the past two decades, it has become accepted that the rotator interval is a distinct anatomic entity that plays an important role in affecting the proper function of the glenohumeral joint. The rotator interval is an anatomic region in the anterosuperior aspect of the glenohumeral joint that represents a complex interaction of the fibers of the coracohumeral ligament, the superior glenohumeral ligament, the glenohumeral joint capsule, and the supraspinatus and subscapularis tendons. As basic science and clinical studies continue to elucidate the precise role of the rotator interval, understanding of and therapeutic interventions for rotator interval pathology also continue to evolve. Lesions of the rotator interval may result in glenohumeral joint contractures, shoulder instability, or in lesions to the long head of the biceps tendon. Long term clinical trials may clarify the results of current surgical interventions and further enhance understanding of the rotator interval. PMID- 17426294 TI - The use of lumbar epidural/transforaminal steroids for managing spinal disease. AB - Lumbar epidural steroid injections are used to manage low back and leg pain (ie, sciatica). Utilization of the procedure is increasing, with Medicare spending for lumbar epidural procedures topping $175 million annually. Few prospective randomized controlled trials have clearly demonstrated the efficacy of epidural steroid injections; many have shown conflicting results. Several studies show favorable short-term outcomes with epidural steroid injection for radicular pain, but less conclusive results are achieved >6 months. Methodologic flaws limit interpretation of results from most scientific studies. As a tool for predicting surgical outcome, epidural spinal injection has been found to have a sensitivity between 65% and 100%, a specificity between 71% and 95%, and a positive predictive value as high as 95% for 1-year surgical outcome. Despite inconclusive evidence, when weighing the surgical alternatives and associated risk, cost, and outcomes, lumbar epidural steroid injections are a reasonable nonsurgical option in select patients. PMID- 17426295 TI - Acute midshaft clavicular fracture. AB - Clavicular fractures represent 2.6% to 5% of all fractures, and middle third fractures account for 69% to 82% of fractures of the clavicle. The junction of the outer and middle third is the thinnest part of the bone and is the only area not protected by or reinforced with muscle and ligamentous attachments. These anatomic features make it prone to fracture, particularly with a fall on the point of the shoulder, which results in an axial load to the clavicle. Optimal treatment of nondisplaced or minimally displaced midshaft fracture is with a sling or figure-of-8 dressing; the nonunion rate is very low. However, when midshaft clavicular fractures are completely displaced or comminuted, and when they occur in elderly patients or females, the risk of nonunion, cosmetic deformity, and poor outcome may be markedly higher. Thus, some surgeons propose surgical stabilization of a complex midshaft clavicular fracture with either plate-and-screw fixation or intramedullary devices. Further randomized, prospective trials are needed to provide better data on which to base treatment decisions. PMID- 17426296 TI - Neurovascular injuries to the athlete's shoulder: Part I. AB - The neurovascular structures traversing the shoulder region can be compromised in a number of ways. Athletes are particularly at risk of neurovascular injury to the shoulder as the result of extreme force and stress on the shoulder girdle. Many such injuries have been described in the literature as cervical radiculitis, spinal accessory nerve injury, long thoracic nerve palsy, burner (stinger) syndrome, and brachial neuritis. A high index of diagnostic acumen and proper selection of clinical assessment and imaging techniques are needed to diagnose such injuries. PMID- 17426299 TI - "O" for opportunity. PMID- 17426300 TI - Update in the pharmacologic treatment of diabetes: focus on insulin detemir, insulin glulisine, and inhaled dry powdered insulin. AB - The prevalence of diabetes mellitus has increased to more than 20 million people in the United States, and current estimates indicate that one third of all Americans born in the year 2000 will develop diabetes mellitus in their lifetime. The need for diabetes therapies offering improved glucose control by mimicking normal physiological properties of glucose metabolism and improving on logistics such as ease of use, self-management, monitoring, and delivery is clear. This article, part 2 of a 3-part series, reviews newer injectable insulin preparations and examines the first-to-market orally inhaled dry powdered insulin (IDPI). The information provided is tailored to diabetes educators and includes mechanism of action, pharmacokinetics, drug interactions, clinical trials, dosage and administration guidelines, side effects, and educational pearls for each insulin discussed. A detailed patient case designed to acquaint the reader with these newer insulin products and provide an understanding of clinical issues to consider when providing diabetes education to patients is included. PMID- 17426301 TI - The new indicator test (Neuropad): a valuable diagnostic tool for small-fiber impairment in patients with type 2 diabetes. AB - PURPOSE: The purpose of this study was to evaluate the new indicator test for sudomotor function (Neuropad) in the diagnosis of small-fiber impairment in patients with type 2 diabetes. METHODS: This study included 123 patients with type 2 diabetes (59 men; mean age, 64.3 +/- 8.6 years; mean diabetes duration, 12 +/- 6.1 years). Sudomotor dysfunction was assessed by means of the new indicator test. Neuropathy was diagnosed by the Neuropathy Disability Score and small-fiber impairment by temperature perception (Tiptherm device) and pain perception (Neurotip). RESULTS: The frequency of sudomotor dysfunction was significantly (P = .001) higher in patients with neuropathy (95%) than in those without neuropathy (30.2%). Sensitivity of the indicator test for neuropathy was 95%, and specificity was 69.8%. Frequency of neuropathy was significantly (P = .018) higher with the indicator test (74.8%) than with conventional clinical examination (65.4%). Sudomotor dysfunction was significantly (P = .001) more frequent in patients with small-fiber impairment (99%) than in those without small-fiber impairment (21.7%). Sensitivity for small-fiber impairment was 99%, and specificity was 78.3%. There was no difference (P = .999) in the frequency of small-fiberimpairment as diagnosed with the indicator test (80.5%) and with clinical examination (81.3%). CONCLUSIONS: The indicator test has a very high sensitivity and specificity for small-fiber impairment in patients with type 2 diabetes. PMID- 17426302 TI - Social support and mortality among older persons with diabetes. AB - PURPOSE: The purpose of this study was to examine the relationship between social support and mortality among older persons with diabetes and the pathways by which social support affects diabetes survival. METHODS: Using data from the Longitudinal Study of Aging cohort 2 baseline (1994) and follow-up (1997-1998 and 1999-2000 surveys), the authors identified 1431 persons aged >or=70 years with diabetes, among whom 387 deaths occurred. Social support was measured by an index with regard to participants' connection with relatives, friends, neighbors, social events, church, and senior centers. Regression analysis was used to find the pathway, and survival analysis was used to find the relationship between social support and mortality. RESULTS: Compared to people with a low level of social support, the risk of death is 41% lower among people with medium levels of support (hazards ratio = 0.59, 0.39-0.91) and 55% lower among those with the highest levels of support (hazards ratio = 0.45, 0.21-0.98). Eight of the 11 regression models demonstrated that the effect of social support on mortality was mediated by both physical and mental health status. CONCLUSIONS: Social support is strongly associated with mortality. Based on findings from this study, social support should be considered an important target for intervention to reduce mortality risk among older adults with diabetes. PMID- 17426303 TI - Treatment and awareness of type 2 diabetes in Beijing, China, compared to New York. AB - PURPOSE: The purpose of this study was to determine awareness and treatment of type 2 diabetes among Beijingers. METHODS: Surveys generated in Mandarin and English were used to poll 75 Beijingers with type 2 diabetes, 29 Beijingers without diabetes, and 23 New Yorkers without diabetes. Beijing data were compared with diabetes statistics on exercise and blood glucose testing frequency from the 2002 New York City Department of Health and Mental Hygiene Community Health Survey and the 2003, 2004, and 2005 New York State Behavioral Risk Factor Surveillance System. RESULTS: The surveyed Beijingers with diabetes used primarily only Western pills to treat their type 2 diabetes, and a smaller percentage used traditional Chinese medicine. Most of the surveyed Beijingers with diabetes wrote that they exercised regularly. Most exercised at least 7 times per week. About half of the surveyed Beijingers with diabetes had no food restrictions. Virtually all of the surveyed Beijingers with diabetes tested their blood glucose, but approximately half of these individuals tested less than 1 time per week. Beijingers with diabetes were less aware than New York respondents who did not have diabetes of what diabetes is and the consequences of poor diabetes treatment. Beijingers who did not have diabetes were less aware than New Yorkers who did not have diabetes of what diabetesis, the symptoms, the causes, the treatments, and the consequences of poor diabetes treatment. CONCLUSIONS: The results indicate that compared to New Yorkers with diabetes, Beijingers with diabetes tended to exercise much more frequently but tested their blood glucose less frequently. With the projected increase of type 2 diabetes in Beijing, more efforts need to be made to increase the frequency of blood glucose testing and basic diabetes awareness. PMID- 17426304 TI - Diabetes problem-solving scale development in an adult, African American sample. AB - PURPOSE: The purpose of this pilot study was to examine psychometric properties of the Diabetes Problem-Solving Scale (DPSS), which was designed to assess how adults with type 2 diabetes approach and manage problems encountered in diabetes self-management. METHODS: Participants were 64 African American adults with type 2 diabetes. The 30-item DPSS and measures of social problem solving, diabetes self-management, and depressive symptoms were administered. Blood samples were collected to measure hemoglobin A1C level. RESULTS: Cronbach alpha for the DPSS total scale was .77 and ranged from .72 to .78 for subscales. Correlations of the DPSS total score and subscale scores with a standardized social problem-solving scale ranged from 0.30 to 0.46 (all P < .01). Higher DPSS total scores, indicating better self-reported diabetes problem solving, were associated with higher medication adherence, more frequent self-monitoring of blood glucose, and lower hemoglobin A1C level. Of the DPSS subscales, Impulsive Style, Negative Transfer of Past Experience/Learning, and Negative Motivation were differentially associated with reduced self-management and disease control. CONCLUSION: The DPSS demonstrated acceptable total scale and subscale internal consistency, construct validity, and predictive validity in this pilot sample. The scale may have utility both in identifying associations between diabetes-related problem solving and self-management and in guiding problem solving interventions to improve self management and control. PMID- 17426305 TI - Health beliefs of Mexican Americans with type 2 diabetes: The Starr County border health initiative. AB - PURPOSE: The purpose of this study was to compare 2 culturally competent diabetes self-management interventions designed for Mexican Americans: an original extended program (24 hours of education, 28 hours of support groups) versus a shorter, more resource-efficient compressed strategy (16 hours of education, 6 hours of support groups). The effects of the interventions on health beliefs are compared. METHODS: The authors recruited 216 persons between 35 and 70 years of age diagnosed with type 2 diabetes for at least 1 year. Intervention groups of 8 participants and 8 support persons were randomly assigned to 1 of the interventions. RESULTS: Mean health belief scores on each subscale improved for both intervention groups. Both intervention groups reported significant improvements in perceptions of control of their diabetes. Improvements in health beliefs were more sustained at 12 months for individuals in the longer, extended program. The health belief subscale control was the most significant predictor of HbA1c levels at 12 months. CONCLUSIONS: Both culturally competent diabetes self management education interventions were effective in promoting more positive health beliefs. These findings on health beliefs indicate a dosage effect of the intervention and support the importance of ongoing contact through support groups to attain more sustainable improvements in health beliefs. PMID- 17426306 TI - Further evaluating the acceptability of group visits in an uninsured or inadequately insured patient population with uncontrolled type 2 diabetes. AB - PURPOSE: The purpose of the study was to evaluate perceptions of care delivered through group visits to disadvantaged patients with type 2 diabetes. METHODS: One hundred eighty-six patients with uncontrolled type 2 diabetes were randomly assigned to receive care in group visits or usual care for 12 months. Their perceptions of the care they received were measured at baseline and 6 and 12 months by the Primary Care Assessment Tool (PCAT), the Diabetes-Specific Locus of Control (DLC) survey, and the Trust in Physician Scale (TPS). RESULTS: Compared to patients in usual care, group visit patients' PCAT scores were higher in the domains of ongoing care (P = .001), community orientation (P < .0001), and cultural competence (P = .022). In addition, group patients had higher scores for the Powerful-Other Health Professional subscale of the DLC survey (P = .010). CONCLUSIONS: Patients assigned to group visits had generally more positive perceptions about their care in the areas of ongoing care, community orientation of care, and cultural competence of care than did those in usual care. The perception that one's health professional is powerful, however, has been associated with a reluctance of patients to make medication changes on their own in previous studies. These findings suggest the need for modification in the way that group visits are conducted to empower and activate patients while still delivering continuous, culturally competent, and community-oriented care. PMID- 17426307 TI - The context of a legacy of diabetes: correlates of self-care behavior. AB - PURPOSE: The purpose of this study is to develop a means to assess recollections of a family member's diabetes self-care behavior and to assess the relationships to a participant's own self-care behavior. Assessing recollections of a family member's self-care behavior may provide insight into what patients learn from family members with diabetes. METHODS: Volunteers with diabetes were recruited from 2 centers in the metropolitan New York/New Jersey area (N = 104) to complete a new pencil-and-paper survey called the Family Summary of Diabetes Self-care Activities Survey, adapted from the widely used, valid and reliable Summary of Diabetes Self-care Activities Survey to assess their recollections about a family member's diabetes self-care behavior. The measure was used to examine the relationships between recollections of a family member's self-care behavior and the participant's own self-care behavior. RESULTS: There was a significant positive association between recollections about family members' dietary behavior with participants' own dietary behavior, indicating that more dietary adherence of the family member is associated with more dietary adherence by the individual. No significant associations were found between the other self-care behaviors (physical activity, glucose monitoring, pill and insulin adherence). CONCLUSION: The relationship of recollections about the self-care of family members to individuals' own self-care provides some empirical support for the concept of multigenerational legacies of diabetes. PMID- 17426308 TI - An uncompromising approach to achieving glycemic goals. PMID- 17426309 TI - Removing barriers to insulin therapy. PMID- 17426310 TI - Practical approaches to insulin therapy. PMID- 17426311 TI - Time to move on. PMID- 17426312 TI - High-fat diets: modeling the metabolic disorders of human obesity in rodents. AB - RESEARCH METHODS AND PROCEDURES: High-fat (HF) diet feeding can induce obesity and metabolic disorders in rodents that resemble the human metabolic syndrome. However, this dietary intervention is not standardized, and the HF-induced phenotype varies distinctly among different studies. The question which HF diet type is best to model the metabolic deterioration seen in human obesity remains unclear. Therefore, in this review, metabolic data obtained with different HF diet approaches are compiled. Both whole-body and organ-specific diet effects are analyzed. RESULTS: On the basis of these results, we conclude that animal fats and omega-6/omega-9-containing plant oils can be used to generate an obese and insulin-resistant phenotype in rodents, whereas fish oil-fed animals do not develop these disorders. DISCUSSION: Looking at the present data, it does not seem possible to define an ideal HF diet, and an exact definition of diet composition and a thorough metabolic characterization of the HF diet effects in a researcher's specific laboratory setting remains essential for metabolic studies with this model. PMID- 17426313 TI - Association of neuropeptide Y receptor Y5 polymorphisms with dyslipidemia in Mexican Americans. AB - We examined the genetic association of neuropeptide Y receptor Y5 (NPY5R) single nucleotide polymorphisms (SNPs) with measures of the insulin resistance (metabolic) syndrome. We genotyped 10 NPY5R SNPs in 439 Mexican American individuals (age=43.3+/-17.3 years and BMI=30.0+/-6.7 kg/m2) distributed across 27 pedigrees from the San Antonio Family Diabetes Study and performed association analyses using the measured genotype approach as implemented in Sequential Oligogenic Linkage Analysis Routines (SOLAR). Minor alleles for five (rs11100493, rs12501691, P1, rs11100494, rs12512687) of the NPY5R SNPs were found to be significantly (p<0.05) associated with fasting plasma triglyceride concentrations and decreased high-density lipoprotein concentrations. In addition, the minor allele for SNP P2 was significantly associated (p=0.031) with a decreased homeostasis model assessment of beta-cell function (HOMA-%beta). Linkage disequilibrium between SNP pairs indicated one haplotype block of five SNPs (rs11100493, rs12501691, P1, rs11100494, rs12512687) that were highly correlated (r2>0.98). These preliminary results provide evidence for association of SNPs in the NPY5R gene with dyslipidemia (elevated triglyceride concentrations and reduced high-density lipoprotein levels) in our Mexican American population. PMID- 17426314 TI - The association between birth weight and visceral fat in middle-age adults. AB - OBJECTIVE: Low birth weight, a proxy for fetal underdevelopment, is associated with increased risk of developing type 2 diabetes during adulthood. Low birth weight is also associated with central obesity, but little is known about the association between birth weight and visceral adiposity. The purpose of this study is to test the hypothesis that lower birth weight is associated with increased amounts of visceral fat in middle-age adults. RESEARCH METHODS AND PROCEDURES: This is an observational study of 91 adults (58 men and 33 women) 40+/-6 years of age (mean+/-standard deviation). Ethnicity was either Japanese American (79%) or non-Hispanic white (21%). Birth weight was obtained from State Departments of Health. Measurements included smoking status, BMI, and visceral (intra-abdominal) fat measured by computed tomography. RESULTS: Visceral fat was not associated with birth weight after adjustment for age, sex, ethnicity, BMI, or smoking status (p=0.76). There was no evidence that the association between birth weight and visceral fat varied by age, sex, or ethnicity. DISCUSSION: We found no evidence that low birth weight is associated with increased visceral fat in middle-age adults. PMID- 17426315 TI - Sex-specific difference in the interconversion of cortisol and cortisone in men and women. AB - OBJECTIVE: Our objective was to demonstrate that the smaller oxoreductase activity of 11beta-HSD1 in women would shift the interconversion of cortisol and cortisone toward cortisone, resulting in a larger amount of generated labeled cortisone in healthy women than in healthy men. RESEARCH METHODS AND PROCEDURES: Using mass spectrometry, the amount of cortisone generated from a continuous infusion (8 am to 6 pm) of stable-labeled cortisol (1alpha,2alpha-d-cortisol) was determined in non-obese and in obese (BMI>35 kg/m2) men and women during steady state conditions (from 2 pm to 6 pm). In this setting, the amount of generated labeled cortisone (expressed as % of the achieved steady-state concentrations of labeled cortisol) reflects the sum of the bi-directional conversion of cortisol into cortisone (and vice versa) by 11beta-hydroxysteroid dehydrogenase. RESULTS: The amount of generated labeled cortisone was higher in men than in women (p<0.0001). This sex difference was higher in obese than in non-obese patients (p=0.0062). CONCLUSIONS: The interconversion of cortisol and cortisone during steady-state conditions is shifted toward cortisol in men as compared with women. This suggests a higher overall oxoreductase activity of 11beta-hydroxysteroid dehydrogenase type 1 in men than in women. This sex-specific difference is maintained in obesity. PMID- 17426316 TI - The efficacy of a technology-based system in a short-term behavioral weight loss intervention. AB - OBJECTIVE: The objective was to examine the efficacy of adding a technology-based program to an in-person, behavioral weight loss intervention. RESEARCH METHODS AND PROCEDURES: Fifty-seven subjects (BMI=33.1+/-2.8 kg/m2; age=41.3+/-8.7 years) participated in a 12-week intervention with random assignment to Standard In Person Behavioral Weight Control Program (SBWP) or Intermittent or Continuous Technology-Based Program (INT-TECH, CON-TECH). SBWP subjects received seven individualized weight loss sessions encouraging dietary and exercise modifications. INT-TECH and CON-TECH subjects received all SBWP components; additionally, these groups used a SenseWear Pro Armband (BodyMedia, Inc.) to monitor energy expenditure and an Internet-based program to monitor eating behaviors. These features were used by INT-TECH subjects during weeks 1, 5, and 9 and CON-TECH subjects weekly throughout the intervention. RESULTS: Intent-to treat analysis revealed weight loss of 4.1+/-2.8 kg, 3.4+/-3.4 kg, and 6.2+/-4.0 kg, for SBWP, INT-TECH, and CON-TECH groups, respectively (CON-TECH>INT-TECH, por=6% of initial body weight (n=359) were randomized to 52 weeks of 1 mg/d MK-0557 or placebo and maintained on a hypocaloric diet (300 kcal below weight maintenance requirements). RESULTS: In randomized patients, the VLCD was associated with an average weight loss of 9.1 kg. After 12 weeks of double-blind treatment, weight began to gradually increase for both placebo- and MK-0557 treated patients. The mean weight change (95% confidence interval) from baseline at the end of the VLCD to Week 52 was +3.1 (2.1, 4.0) and +1.5 (0.5, 2.4) kg for patients treated with placebo and MK-0557, respectively. The difference of 1.6 kg between the two groups was significant (p=0.014). Secondary endpoints, such as blood pressure, lipid profile, insulin, and leptin, as well as waist circumference and quality-of-life measurements, did not show significant differences between MK-0557 and placebo treatments. DISCUSSION: Although the difference in weight regain between placebo- and MK-0557-treated patients was statistically significant, the magnitude of the effect was small and not clinically meaningful. Antagonism of the neuropeptide Y Y5 receptor is not an efficacious treatment strategy for reducing weight regain after VLCD. PMID- 17426326 TI - Wise Mind project: a school-based environmental approach for preventing weight gain in children. AB - OBJECTIVE: The Wise Mind pilot study compared the efficacy of an environmental approach for prevention of inappropriate weight gain in children with an active control condition that used an environmental approach for modifying expectancies related to the use of alcohol, tobacco, and drugs. RESEARCH METHODS AND PROCEDURES: A total of 670 second to sixth grade students from four schools were enrolled in the study. The study spanned 2 academic years, and 586 students were available for evaluation at the end of the study. Two schools were randomly assigned to each treatment arm. The environmental approach for weight gain prevention focused on modification of eating habits and physical activity, and the active control group focused on modification of expectancies related to substance use. RESULTS: Using an intention to treat design, the study found no differences in weight gain prevention between the two interventions. The weight gain prevention program was associated with reduction of total caloric intake, reduction of dietary fat intake, reduction of protein intake, and increased physical activity in comparison with the active control group and relative to baseline. These changes in food intake were attributed to changes in food selections that resulted from modification of school cafeteria menus and food preparation. DISCUSSION: The Wise Mind school-based weight gain prevention program induced behavioral changes in healthy eating and physical activity but did not induce significant changes in body weight in comparison with the control arm. Recommendations for future research are provided. PMID- 17426327 TI - Field validation of the MTI Actigraph and BodyMedia armband monitor using the IDEEA monitor. AB - OBJECTIVE: Accelerometers offer considerable promise for improving estimates of physical activity (PA) and energy expenditure (EE) in free-living subjects. Differences in calibration equations and cut-off points have made it difficult to determine the most accurate way to process these data. The objective of this study was to compare the accuracy of various calibration equations and algorithms that are currently used with the MTI Actigraph (MTI) and the Sensewear Pro II (SP2) armband monitor. RESEARCH METHODS AND PROCEDURES: College-age participants (n=30) wore an MTI and an SP2 while participating in normal activities of daily living. Activity patterns were simultaneously monitored with the Intelligent Device for Estimating Energy Expenditure and Activity (IDEEA) monitor to provide an accurate estimate (criterion measure) of EE and PA for this field-based method comparison study. RESULTS: The EE estimates from various MTI equations varied considerably, with mean differences ranging from -1.10 to 0.46 METS. The EE estimates from the two SP2 equations were within 0.10 METS of the value from the IDEEA. Estimates of time spent in PA from the MTI and SP2 ranged from 34.3 to 107.1 minutes per day, while the IDEEA yielded estimates of 52 minutes per day. DISCUSSION: The lowest errors in estimation of time spent in PA and the highest correlations were found for the new SP2 equation and for the recently proposed MTI cut-off point of 760 counts/min (Matthews, 2005). The study indicates that the Matthews MTI cut-off point and the new SP2 equation provide the most accurate indicators of PA. PMID- 17426328 TI - Obesity history as a predictor of walking limitation at old age. AB - OBJECTIVE: To study whether walking limitation at old age is determined by obesity history. RESEARCH METHODS AND PROCEDURES: In a retrospective longitudinal study based on a representative sample of the Finnish population of 55 years and older (2055 women and 1337 men), maximal walking speed, body mass, and body height were measured in a health examination. Walking limitation was defined as walking speed<1.2 m/s or difficulty in walking 0.5 km. Recalled height at 20 years of age and recalled weight at 20, 30, 40, and 50 years of age were recorded. RESULTS: Subjects who had been obese at the age of 30, 40, or 50 years had almost a 4-fold higher risk of walking limitation compared to non-obese. Obesity duration increased the age- and gender-adjusted risk of walking limitation among those who had been obese since the age of 50 (odds ratio, 4.33; 95% confidence interval, 2.59 to 7.23, n=114), among the obese since the age of 40 [6.01 (2.55 to 14.14), n=39], and among the obese since the age of 30 [8.97 (3.06 to 26.29), n=14]. The risk remained elevated even among those who had previously been obese but lost weight during their midlife or late adulthood [3.15 (1.63 to 6.11), n=71]. DISCUSSION: Early onset of obesity and obesity duration increased the risk of walking limitation, and the effect was only partially mediated through current BMI and higher risk of obesity-related diseases. Preventing excess weight gain throughout one's life course is an important goal in order to promote good health and functioning in older age. PMID- 17426329 TI - Randomized trial of a multifaceted commercial weight loss program. AB - OBJECTIVE: To test whether a commercial weight loss program promotes greater weight loss in overweight or obese women compared with control conditions and to describe the effect on plasma lipids, carotenoids, hormones, and fitness. RESEARCH METHODS AND PROCEDURES: Overweight or obese women were randomized to commercial weight loss program or control conditions (n=35 each). RESULTS: At randomization, participants were 41.1 (11.4) (mean [standard deviation]) years, BMI 34.0 (3.5) kg/m2, and weight 92.0 (11.1) kg. At 6 months, change in weight by intent-to-treat (ITT) analysis was -7.2 (6.7) kg and -7.8% (7.2%) in the intervention group vs. -0.3 (3.9) kg and -0.3% (4.5%) in the control group (n=35 for each; p<0.01). One-year ITT analysis revealed significantly greater change in weight, percent weight, BMI, and waist and hip circumferences in the intervention vs. control group. Completers at 1 year exhibited change in weight of -7.3 (10.4) kg for the intervention group (n=32) vs. -0.7 (5.6) kg for controls (n=33) (p<0.01), and -7.8% (11.1%) weight change for the intervention group vs. -0.7% (6.2%) for controls (p<0.01). High-density lipoprotein (HDL) cholesterol concentration increased significantly in the intervention group. Fasting serum insulin decreased in the intervention but increased in the control group at 6 months (p<0.01), remaining different at 1 year (p=0.05). DISCUSSION: The commercial program successfully facilitated weight loss, which was notably maintained at 1 year, and promoted favorable changes in plasma lipid and hormone concentrations. PMID- 17426330 TI - Physical activity-related energy expenditure with the RT3 and TriTrac accelerometers in overweight adults. AB - OBJECTIVE: The objective was to evaluate two accelerometers, the RT3 and the TriTrac-R3D for their ability to produce estimates of physical activity-related energy expenditure (PAEE) in overweight/obese adults. RESEARCH METHODS AND PROCEDURES: PAEE estimates from both accelerometers were obtained in two experiments. In Experiment 1, 13 overweight/obese subjects (BMI 34.2+/-6.4 kg/m2) were monitored over 2 weeks in everyday life, PAEE being simultaneously measured by the doubly labeled water method (DLW). In Experiment 2, 8 overweight/obese subjects (BMI 34.3+/-5.0 kg/m2) and 10 normal-weight subjects (BMI 20.8+/-2.1 kg/m2) were monitored during a treadmill walking protocol, PAEE being simultaneously measured by indirect calorimetry. RESULTS: In Experiment 1, there was no significant difference between methods in mean PAEE (DLW: 704+/-223 kcal/d, RT3: 656+/-140 kcal/d, TriTrac-R3D 624+/-419 kcal/d). The relative difference between methods (accelerometer vs. DLW) was -17.1%+/-16.7% for the RT3 and -20.0+/-44.6% for the TriTrac-R3D. Correlation for PAEE between RT3 and DLW was higher than between TriTrac-R3D and DLW (r=0.67, p<0.05 and r=0.36, p=0.25, respectively). The 95% confidence interval (CI) (kcal/d) of the mean difference between methods was large, amounting to -385 to 145 for the RT3 and -887 to 590 for the TriTrac-R3D. In Experiment 2, both accelerometers were sensitive to the changes in treadmill speed, with no significant difference in mean PAEE between methods in overweight/obese subjects. CONCLUSIONS: Although both accelerometers did not provide accurate estimates of PAEE at individual levels, the data suggest that RT3 has the potential to assess PAEE at group levels in overweight/obese subjects. PMID- 17426331 TI - Short-term effects of a "health-at-every-size" approach on eating behaviors and appetite ratings. AB - OBJECTIVE: To assess the effects of a "Health-At-Every-Size" (HAES) intervention on eating behaviors and appetite ratings in 144 premenopausal overweight women. RESEARCH METHODS AND PROCEDURES: Women were randomly assigned to one of the 3 groups: HAES group, social support (SS) group, and control group (N = 48 in each group). Interventions were conducted over a 4-month period, and measurements were taken before and after this period. Eating behaviors (cognitive dietary restraint, disinhibition, and susceptibility to hunger) were evaluated by the Three-Factor Eating Questionnaire. Appetite ratings (desire to eat, hunger, fullness, and prospective food consumption) were assessed by visual analogue scales before and after a standardized breakfast. RESULTS: More important decreases in susceptibility to hunger and external hunger were observed in the HAES group when compared with the SS group (p=0.05, for susceptibility to hunger) and the control group (p=0.02 and p=0.005, for susceptibility to hunger and external hunger, respectively). In addition, women from the HAES group had more important decreases in postprandial area under the curve for desire to eat (p=0.02) and hunger (p=0.04) when compared with the control group. The change in the desire to eat noted in the HAES group was also different from the one observed in SS group (p=0.02). Women from the HAES group experienced significant weight loss at 4 months (-1.6+/-2.5 kg, p<0.0001), which did not differ significantly from the SS and control groups (p=0.09). An increase in flexible restraint was significantly related to a greater weight loss in both HAES and SS groups (r=-0.39, p<0.01; and r=-0.37, p<0.05, respectively). A decrease in habitual susceptibility to disinhibition was also associated with a greater weight loss in HAES and control groups (r=0.31, p<0.05; and r=0.44, p<0.05, respectively). DISCUSSION: These results suggest that a HAES intervention could have significant effects on eating behaviors and appetite ratings in premenopausal overweight women, when compared with an SS intervention or a control group. PMID- 17426332 TI - Dietary fat and weight gain among women in the Nurses' Health Study. AB - OBJECTIVE: To assess the association of dietary fat and weight gain among adult women and to investigate whether offspring of overweight parents have a greater predisposition to weight gain due to intake of dietary fat. RESEARCH METHODS AND PROCEDURES: This was an 8-year follow-up of 41,518 women in the Nurses' Health Study (NHS), a population-based, prospective cohort. The women were 41 to 68 years of age, free of cardiovascular disease, cancer, and diabetes in 1986 when "baseline" weight and diet were assessed. Eight years later (1994), changes in weight and dietary intake were assessed. Linear regression models were used to relate change in weight to fat intake and change in fat intake, using the percentage of energy from carbohydrate as the comparison, adjusted for age, BMI in 1986, leisure time physical activity, time spent sitting, percent of calories from protein, and change in percentage of calories from protein. RESULTS: Overall, there was a weak positive association between total fat intake (beta=0.11) and weight gain. Increases in monosaturated and polyunsaturated fat were not associated with weight gain, but increases in animal fat, saturated fat, and trans fat had a positive association with weight change. There was not strong evidence of effect modification by parental weight status (p=0.7 to 0.8 for percentage of calories from total fat, animal fat, and vegetable fat); however, the associations were stronger among the overweight compared with leaner women (p<0.05 for percentage of calories from each type of fat). Among overweight women, for every one percentage increase in percentage of calories from trans fat, women gained an additional 2.3 lb (95% confidence interval, 1.80 to 2.86). CONCLUSION: Our results show that, overall, percent of calories from fat has only a weak positive association with weight gain; however, percentage of calories from animal, saturated, and trans fat has stronger associations. There was no clear evidence that the diet-weight gain association was stronger among offspring of overweight parents, but dietary fat was associated with greater weight gain among overweight women. PMID- 17426333 TI - Secular trends in childhood obesity in Denmark during 50 years in relation to economic growth. AB - OBJECTIVE: Our aim was to examine whether secular trends in childhood overweight and obesity during five decades could be explained by economic growth. RESEARCH METHODS AND PROCEDURES: Annual measurements of height and weight were available for all children born between 1930 and 1983 attending primary school in the Copenhagen Municipality: 165,389 boys and 163,609 girls from the age of 7 through 13 years. After computerization, we calculated BMI (kg/m2) and estimated the prevalence of overweight and obesity, according to international age- and gender specific criteria, by year of birth and of measurement, and separately by each age group and gender. Economic growth was indicated by the Gross National Product and the overall consumption per capita, adjusted for inflation. RESULTS: The prevalence of overweight occurred in phases: an increase from 1930 until the 1950s, followed by a plateau period between the 1950s and the 1960s and a steep increase thereafter. This pattern was apparent across all age groups and in both genders. Obesity trends showed a similar phase pattern; the prevalence remained relatively stable from 1930 until the 1940s, increased until the mid-1950s, followed by a plateau until 1965, and thereafter a second steep increase. Obesity trends were similar among boys across all age groups, although only among girls from 11 to 13 years of age. In both genders, increments were most pronounced in the upper BMI percentiles. After stagnation until 1947, the economic growth indicators showed a steady increase; i.e., after the first increase started in overweight and obesity, whether analyzed by year of birth or year of measurement, there were no indications of phases in the rise thereafter. DISCUSSION: Prevalence of overweight and obesity among Danish children rose in phases, which were not paralleled by trends in economic growth. The macroeconomic growth indicators seem inappropriate as proxies for the environmental exposures that have elicited the obesity epidemic. PMID- 17426334 TI - Trends in pre-pregnancy obesity in nine states, 1993-2003. AB - OBJECTIVE: Pre-pregnancy obesity poses risks to both pregnant women and their infants. This study used a large population-based data source to examine trends, from 1993 through 2003, in the prevalence of pre-pregnancy obesity among women who delivered live infants. RESEARCH METHODS AND PROCEDURES: Data from the Pregnancy Risk Assessment Monitoring System in nine states were analyzed for trends in pre-pregnancy obesity (BMI>29.0 kg/m2) overall and by maternal demographic and behavioral characteristics. Pre-pregnancy BMI was calculated from self-reported weight and height on questionnaires administered after delivery, and demographic characteristics were taken from linked birth certificates. The sample of 66,221 births was weighted to adjust for survey design, non-coverage, and non-response, and it is representative of all women delivering a live birth in each particular state. The sampled births represented 18.5% of all births in the United States. RESULTS: Pre-pregnancy obesity increased 69.3% during the study period, from 13.0% in 1993 to 1994 to 22.0% in 2002 to 2003. The percentage increase ranged from 45% to 105% for individual states. Subgroups of women with the highest prevalence of obesity in 2002 to 2003 were those who were 20 to 29 years of age, black, had three or more children, had a high school education, enrolled in Women, Infants, and Children, or were non-smokers. However, all subgroups of women examined experienced at least a 43% increase in pre-pregnancy obesity over this time period. DISCUSSION: The prevalence of pre-pregnancy obesity is increasing among women in these nine states, and this trend has important implications for all stages of reproductive health care. PMID- 17426335 TI - A comparison of adiposity measures as predictors of all-cause mortality: the Melbourne Collaborative Cohort Study. AB - OBJECTIVE: Our goal was to examine five different measures of adiposity as predictors of all-cause mortality. RESEARCH METHODS AND PROCEDURES: Subjects were 16,969 men and 24,344 women enrolled between 1990 and 1994 in the Melbourne Collaborative Cohort Study (27 to 75 years of age). There were 2822 deaths over a median follow-up period of 11 years. BMI, waist circumference, and waist-to-hip ratio were obtained from direct anthropometric measurements. Fat mass and percentage fat were estimated by bioelectric impedance analysis. RESULTS: Comparing the top quintile with the second quintile, for men there was an increased risk of between 20% and 30% for all-cause mortality associated with each of the anthropometric measures. For women, there was an increased risk of 30% (95% confidence interval for hazard ratio, 1.1-1.6) observed for waist circumference and 50% (1.2-1.8) for waist-to-hip ratio, but little or no increased risk for BMI, fat mass, and percentage fat. Waist-to-hip ratio was positively and monotonically associated with all-cause mortality for both men and women. There was a linear association between waist circumference and all-cause mortality for men, whereas a U-shaped association was observed for women. DISCUSSION: Measures of central adiposity were better predictors of mortality in women in the Melbourne Collaborative Cohort Study compared with measures of overall adiposity. We recommend measuring waist and hip circumferences in population studies investigating the risk of all-cause mortality associated with obesity. The use of additional measures such as bioelectric impedance is not justified for this outcome. PMID- 17426336 TI - Higher insulin, triglycerides, and blood pressure with greater trunk fat in Tanner 1 Chinese. AB - OBJECTIVE: The aims of this study were to investigate the body fat distribution pattern in prepubertal Chinese children and to investigate the relationship between central fat distribution and specific biomarkers of cardiovascular disease. RESEARCH METHODS AND PROCEDURES: The study was conducted in an urban Mainland Chinese (Jinan, Shandong) sample of children using a cross-sectional design. Pubertal status was determined by Tanner criteria. Measurements included weight, height, waist circumference, DXA measures of total body fat and trunk fat; fasting serum measures of glucose, insulin, triglyceride, cholesterol, high density lipoprotein-cholesterol; and systolic and diastolic blood pressure. Multiple regression models were developed with the biomarkers of cardiovascular risk factor as the dependent variables, and adjustments were made for significant covariates, including sex, age, height, weight, waist circumference, total body fat, trunk fat, and interactions. RESULTS: A total of 247 healthy prepubertal subjects were studied. After co-varying for age, weight, height, and extremity fat (the sum of arm fat and leg fat), girls had greater trunk fat than boys (p<0.0001, R2 for model=0.95). Insulin and triglyceride were positively related to central fat measured by DXA-trunk fat (p<0.05) but not related to the waist circumference. In the blood pressure model, waist circumference was a significant predictor of both systolic blood pressure and diastolic blood pressure, while DXA trunk fat was associated with diastolic blood pressure only. Significant interactions between sex and trunk fat, and sex and total fat, were found in relation to diastolic blood pressure. DISCUSSION: In prepubertal Chinese children, greater trunk fat was significantly associated with higher insulin and triglyceride in boys and girls and was associated with higher diastolic blood pressure in boys only. PMID- 17426337 TI - The association between changes in height and obesity in Chilean preschool children: 1996-2004. AB - OBJECTIVES: To establish the association between changes in height and prevalence of obesity between 1996 and 2004 in Chilean preschool children. RESEARCH METHODS AND PROCEDURES: Children registered in the Junta Nacional de Jardines Infantiles (JUNJI) preschool program were routinely measured three times a year and in consecutive years. Two indices [weight-for-height z-scores (WHZ) and BMI centiles] were used to define obesity. Height-for-age z-scores (HAZ) were used to define stature. Generalized estimating equations were used to describe the relationship between stature and obesity while controlling for repeated measurements in children. RESULTS: The prevalence of obesity in Chilean preschool children has remained relatively constant over the past 9 years when either index is used. The prevalence of stunted (HAZ2) has increased. Using WHZ to define obesity, stunted children do not seem to be at increased risk of obesity when compared with children of normal height. Tall children were strongly protected from obesity when either criterion was used to define obesity. DISCUSSION: The lack of association between stunting and obesity using WHZ>or=2 likely reflects the fact that Chile is in the post-transitional phase of the nutrition transition, and stunting is no longer a risk factor for obesity; however, the associations observed between stunting and BMI suggest that either WHZ or BMI, or both, are inaccurate criteria to define overweight. CONCLUSIONS: There is a unique relationship between stature and obesity in preschool children that is different from that observed in older children in the same population. PMID- 17426338 TI - Association of childhood sexual abuse with obesity in a community sample of lesbians. AB - OBJECTIVE: Our goal was to examine the association between childhood sexual abuse (CSA) and obesity in a community-based sample of self-identified lesbians. RESEARCH METHODS AND PROCEDURES: A diverse sample of women who self-identified as lesbian was recruited from the greater Chicago metropolitan area. Women (n=416) were interviewed about sexual abuse experiences that occurred before the age of 18. Self-reported height and weight were used to calculate BMI and categorize women as normal-weight (<25.0 kg/m2), overweight (25.0 to 29.9 kg/m2), obese (30.0 to 39.9 kg/m2), or severely obese (>or=40 kg/m2). The relationship between CSA and BMI was examined using multinomial logistic regression analysis. RESULTS: Overall, 31% of women in the sample reported CSA, and 57% had BMI>or=25.0 kg/m2. Mean BMI was 27.8 (+/-7.2) kg/m2 and was significantly higher among women who reported CSA than among those who did not report CSA (29.4 vs. 27.1, p<0.01). CSA was significantly related to weight status; 39% of women who reported CSA compared with 25% of women who did not report CSA were obese (p=0.004). After adjusting for age, race/ethnicity, and education, women who reported CSA were more likely to be obese (odds ratio, 1.9; 95% confidence interval, 1.1-3.4) or severely obese (odds ratio, 2.3; 95% confidence interval, 1.1-5.2). DISCUSSION: Our findings, in conjunction with the available literature, suggest that CSA may be an important risk factor for obesity. Understanding CSA as a factor that may contribute to weight gain or act as a barrier to weight loss or maintenance in lesbians, a high-risk group for both CSA and obesity, is important for developing successful obesity interventions for this group of women. PMID- 17426339 TI - Relationship of visceral adiposity to cardiovascular disease risk factors in black and white teens. AB - OBJECTIVE: We tested the hypothesis that visceral adiposity, compared with general adiposity, would explain more of the variance in cardiovascular disease (CVD) risk factors. RESEARCH METHOD AND PROCEDURES: Subjects were 464 adolescents (238 black and 205 girls). Adiposity measures included visceral adipose tissue (VAT; magnetic resonance imaging), percent body fat (%BF; DXA), BMI, and waist girth (anthropometry). CVD risk factors were fasting insulin, fibrinogen, total to high-density lipoprotein-cholesterol ratio, triglycerides (TGs), systolic blood pressure, and left ventricular mass indexed to height2.7. RESULTS: After adjustment for age, race, and sex, all adiposity indices explained significant proportions of the variance in all of the CVD risk factors; %BF tended to explain more variance than VAT. Regression models that included both %BF and VAT found that both indices explained independent proportions of the variance only for total to high-density lipoprotein-cholesterol ratio. For TGs, the model that included both %BF and VAT found that only VAT was significant. For systolic blood pressure and left ventricular mass indexed to height2.7, anthropometric measures explained more of the variance than VAT and %BF. DISCUSSION: The hypothesis that visceral adiposity would explain more variance in CVD risk than general adiposity was not supported in this relatively large sample of black and white adolescents. Only for TGs did it seem that VAT was more influential than %BF. Perhaps the deleterious effect of visceral adiposity becomes greater later in life as it increases in proportion to general adiposity. PMID- 17426340 TI - Abdominal obesity and coronary heart disease in Thai men. AB - OBJECTIVE: To determine the association of four simple anthropometric indices with coronary heart disease (CHD) in Thai men, and to determine the optimal cut off points for each index in the prediction of CHD. RESEARCH METHODS AND PROCEDURES: This is a cohort study with 17 years of follow-up. A total of 2536 male employees from the Electricity Generating Authority of Thailand 35 to 59 years of age at baseline were included in the study. Height, weight, waist circumference, and hip circumference were measured to generate BMI, waist circumference (WC), waist-to-hip ratio (WHR), and waist-to-height ratio (WHtR). Cox regression models were used to estimate hazard ratios by thirds of each index. Receiver operating characteristic curves were used to assess discrimination of CHD. RESULTS: WHtR was most strongly associated with CHD events in Thai men. The age-adjusted hazard ratio for those in the highest, compared with the lowest, third was 2.89 (1.37, 6.11). Although WHtR had the largest area under the receiver operating characteristic curve (AUC) with the optimal cut-off estimated to be 0.51 (sensitivity, 55%; specificity, 61%), no statistically significant difference (p>0.10) was found between the AUC for WHtR and that for the other three indices. CONCLUSION: WHtR is, marginally, the best of the four indices considered to predict CHD events in Thai men. PMID- 17426341 TI - Birthplace, years of residence in the United States, and obesity among Mexican American adults. AB - OBJECTIVE: To evaluate the association between birthplace (Mexico or U.S.) and obesity in men and women and to analyze the relationship between duration of U.S. residency and prevalence of obesity in Mexican immigrants. RESEARCH METHODS AND PROCEDURES: We used cross-sectional data from 7503 adults of Mexican descent residing in Harris County, TX, to evaluate the relationships among BMI, birthplace, and years of residency in the U.S., controlling for demographic characteristics, physical activity level, and acculturation level. RESULTS: U.S. born adults had an increased risk (between 34% and 65%) of obesity compared with their Mexican-born counterparts. After controlling for recognized confounders and risk factors, this association was maintained in the highly acculturated only. Among highly acculturated obese U.S.-born men, 6% of the cases were attributable to the joint effect of birthplace and acculturation; in women, this proportion was 25%. Among Mexican-born women, there was an increasing trend in mean BMI with increasing duration of residency in the U.S.. Compared with immigrants who had lived in the U.S. for <5 years, Mexican-born women who had resided in the U.S. for >or=15 years had an adjusted BMI mean difference of 2.12 kg/m2 (95% confidence interval, 1.53-2.72). DISCUSSION: Mexican-born men and women have a lower risk of obesity than their U.S.-born counterparts, but length of U.S. residency among immigrants, especially in women, is directly associated with risk of obesity. Development of culturally specific interventions to prevent obesity in recent immigrants may have an important public health effect in this population. PMID- 17426342 TI - Effect of retirement on eating out and weight change: an analysis of gender differences. AB - OBJECTIVES: The objectives were to understand how the retirement decisions of older Americans influence household food consumption patterns by gender and, in turn, to examine the impact of the change in food consumption on weight. RESEARCH METHODS AND PROCEDURES: This study used five waves of the Health and Retirement Study (1992 to 2002; n=28,117). Participants were 50 to 71 years old during the study period. We used longitudinal regression analyses controlling for health events, spousal factors, socioeconomic factors, and individual fixed effects over time. RESULTS: Retirement of the individual and of his/her spouse reduced the individual's monthly spending on eating out by $10 and $7 on average, respectively, but did not change household spending on food at home. The wife's, but not the husband's, retirement decreased the spouse's spending on eating out by $13/mo. Spending on eating out was a significant but weak (0.003BMI/$) predictor of weight gain. DISCUSSION: The decrease in spending on eating out after retirement, particularly women's, suggests that people eat out less when they have more time for food preparation at home. However, increases in other risks of weight gain with retirement, such as physical inactivity, could counteract the effects of eating out less. PMID- 17426344 TI - Meeting report: tumor biology--how signaling processes translate to therapy. AB - On the occasion of the 10th Joint Meeting "Signal Transduction-Receptors, Mediators and Genes" in November 2006, the Signal Transduction Society (STS) focused on the holy grail of translational research, that is, how basic signaling research might be transferred into cancer therapy. Participants heard of blockbusters and setbacks and learned of new therapeutic approaches that concentrate on distinct signaling pathways at a more individual level. Thus, the young STS community and the 300 meeting participants left with an optimistic outlook and the notion that the translational task will keep scientists busy all over the world for a while. PMID- 17426345 TI - Metabolic targeting as an anticancer strategy: dawn of a new era? AB - As a result of a spectrum of mitochondrial defects, tumor cells often preferentially use glycolysis to generate adenosine triphosphate (ATP), even in the presence of oxygen, a phenomenon known as aerobic glycolysis, or the "Warburg effect." Dichloroacetate (DCA) is an inhibitor of mitochondrial pyruvate dehydrogenase kinase (PDK), which inhibits pyruvate dehydrogenase (PDH), a gatekeeping enzyme for the entry of pyruvate into the mitochondrial tricarboxylic acid (TCA) cycle. In mice, DCA treatment appears to reactivate mitochondrial respiration in tumor cells, induces their selective killing, and suppresses cancer growth. These observations provide intriguing insights into the plasticity of tumor metabolism that may offer new opportunities for therapeutic intervention. PMID- 17426346 TI - Sequestration and segregation of receptor kinases in epithelial cells: implications for ErbB2 oncogenesis. AB - Cell behaviors are regulated by signaling pathways triggered by the activation of cell surface receptors. A key aspect of receptor signaling is the location of these receptors relative to their ligands and to other receptors, particularly in epithelia, whose cells are polarized by tight junction barriers into apical and basolateral membrane regions. In polarized epithelia, the co-receptor ErbB2 is often localized to the apical surface by its intramembrane ligand Muc4, thus segregating it from its partner ErbB3, which is sequestered at the lateral surface, co-localized with cadherin junctions. The ErbB2-ErbB3 receptor heterodimer, when activated, is a potent stimulator of cell proliferation; thus, the segregation mechanism helps maintain these cells in a differentiated state. Similarly, epidermal growth factor, the ligand for ErbB1, which is present in the apical fluid of some epithelia, is segregated from its receptor by the tight junction barrier. Loss of cell polarity and the tight junction barrier facilitates the interaction of ErbB2 with the hemidesmosome integrin alpha(6)beta(4). This integrin acts as a docking site for signaling pathways to promote cell proliferation and further disruption of cell junctions. The ultimate dissolution of tight junctions may result from activation of transforming growth factor-beta receptors, one subunit of which is directly associated with the junction. This activation triggers degradation of critical tight junction components. These sequestration and segregation phenomena provide a model by which overexpression of the ErbB2 receptor kinase may trigger oncogenesis by initiating junction breakdown. Equally important, these mechanisms may act as a sensor for epithelial damage that can activate repair mechanisms. PMID- 17426347 TI - Rhythmic auditory stimulation improves gait more than NDT/Bobath training in near ambulatory patients early poststroke: a single-blind, randomized trial. AB - OBJECTIVES: The effectiveness of 2 different types of gait training in stroke rehabilitation, rhythmic auditory stimulation (RAS) versus neurodevelopmental therapy (NDT)/Bobath- based training, was compared in 2 groups of hemiparetic stroke patients over a 3-week period of daily training (RAS group, n = 43; NDT/Bobath group =35). METHODS: Mean entry date into the study was 21.3 days poststroke for the RAS group and 22.3 days for the control group. Patients entered the study as soon as they were able to complete 5 stride cycles with handheld assistance. Patients were closely equated by age, gender, and lesion site. Motor function in both groups was pre-assessed by the Barthel Index and the Fugl-Meyer Scales. RESULTS: Pre- to posttest measures showed a significant improvement in the RAS group for velocity (P = .006), stride length (P = .0001), cadence (P = .0001) and symmetry (P = .0049) over the NDT/Bobath group. Effect sizes for RAS over NDT/Bobath training were 13.1 m/min for velocity, 0.18 m for stride length, and 19 steps/min for cadence. CONCLUSIONS: The data show that after 3 weeks of gait training, RAS is an effective therapeutic method to enhance gait training in hemiparetic stroke rehabilitation. Gains were significantly higher for RAS compared to NDT/Bobath training. PMID- 17426348 TI - Loss of cardiac tetralinoleoyl cardiolipin in human and experimental heart failure. AB - The mitochondrial phospholipid cardiolipin is required for optimal mitochondrial respiration. In this study, cardiolipin molecular species and cytochrome oxidase (COx) activity were studied in interfibrillar (IF) and subsarcolemmal (SSL) cardiac mitochondria from Spontaneously Hypertensive Heart Failure (SHHF) and Sprague-Dawley (SD) rats throughout their natural life span. Fisher Brown Norway (FBN) and young aortic-constricted SHHF rats were also studied to investigate cardiolipin alterations in aging versus pathology. Additionally, cardiolipin was analyzed in human hearts explanted from patients with dilated cardiomyopathy. A loss of tetralinoleoyl cardiolipin (L(4)CL), the predominant species in the healthy mammalian heart, occurred during the natural or accelerated development of heart failure in SHHF rats and humans. L(4)CL decreases correlated with reduced COx activity (no decrease in protein levels) in SHHF cardiac mitochondria, but with no change in citrate synthase (a matrix enzyme) activity. The fraction of cardiac cardiolipin containing L(4)CL became much lower with age in SHHF than in SD or FBN mitochondria. In summary, a progressive loss of cardiac L(4)CL, possibly attributable to decreased remodeling, occurs in response to chronic cardiac overload, but not aging alone, in both IF and SSL mitochondria. This may contribute to mitochondrial respiratory dysfunction during the pathogenesis of heart failure. PMID- 17426350 TI - Unobtrusive integration of data management with fMRI analysis. AB - This note describes a software utility, called X-batch which addresses two pressing issues typically faced by functional magnetic resonance imaging (fMRI) neuroimaging laboratories (1) analysis automation and (2) data management. The first issue is addressed by providing a simple batch mode processing tool for the popular SPM software package (http://www.fil.ion. ucl.ac.uk/spm/; Welcome Department of Imaging Neuroscience, London, UK). The second is addressed by transparently recording metadata describing all aspects of the batch job (e.g., subject demographics, analysis parameters, locations and names of created files, date and time of analysis, and so on). These metadata are recorded as instances of an extended version of the Protege-based Experiment Lab Book ontology created by the Dartmouth fMRI Data Center. The resulting instantiated ontology provides a detailed record of all fMRI analyses performed, and as such can be part of larger systems for neuroimaging data management, sharing, and visualization. The X-batch system is in use in our own fMRI research, and is available for download at http://X-batch.sourceforge.net/. PMID- 17426351 TI - The Extensible Neuroimaging Archive Toolkit: an informatics platform for managing, exploring, and sharing neuroimaging data. AB - The Extensible Neuroimaging Archive Toolkit (XNAT) is a software platform designed to facilitate common management and productivity tasks for neuroimaging and associated data. In particular, XNAT enables qualitycontrol procedures and provides secure access to and storage of data. XNAT follows a threetiered architecture that includes a data archive, user interface, and middleware engine. Data can be entered into the archive as XML or through data entry forms. Newly added data are stored in a virtual quarantine until an authorized user has validated it. XNAT subsequently maintains a history profile to track all changes made to the managed data. User access to the archive is provided by a secure web application. The web application provides a number of quality control and productivity features, including data entry forms, data-type-specific searches, searches that combine across data types, detailed reports, and listings of experimental data, upload/download tools, access to standard laboratory workflows, and administration and security tools. XNAT also includes an online image viewer that supports a number of common neuroimaging formats, including DICOM and Analyze. The viewer can be extended to support additional formats and to generate custom displays. By managing data with XNAT, laboratories are prepared to better maintain the long-term integrity of their data, to explore emergent relations across data types, and to share their data with the broader neuroimaging community. PMID- 17426352 TI - Toward a workbench for rodent brain image data: systems architecture and design. AB - We present a novel system for storing and manipulating microscopic images from sections through the brain and higher-level data extracted from such images. The system is designed and built on a three-tier paradigm and provides the research community with a web-based interface for facile use in neuroscience research. The Oracle relational database management system provides the ability to store a variety of objects relevant to the images and provides the framework for complex querying of data stored in the system. Further, the suite of applications intimately tied into the infrastructure in the application layer provide the user the ability not only to query and visualize the data, but also to perform analysis operations based on the tools embedded into the system. The presentation layer uses extant protocols of the modern web browser and this provides ease of use of the system. The present release, named Functional Anatomy of the Cerebro Cerebellar System (FACCS), available through The Rodent Brain Workbench (http:// rbwb.org/), is targeted at the functional anatomy of the cerebro-cerebellar system in rats, and holds axonal tracing data from these projections. The system is extensible to other circuits and projections and to other categories of image data and provides a unique environment for analysis of rodent brain maps in the context of anatomical data. The FACCS application assumes standard animal brain atlas models and can be extended to future models. The system is available both for interactive use from a remote web-browser client as well as for download to a local server machine. PMID- 17426353 TI - Voronoi analysis uncovers relationship between mosaics of normally placed and displaced amacrine cells in the thraira retina. AB - Although neuronal dynamics is to a high extent a function of synapse strength, the spatial distribution of neurons is also known to play an important role, which is evidenced by the topographical organization of the main stations of the visual system: retina, lateral geniculate nucleus, and cortex. The coexisting systems of normally placed and displaced amacrine cells in the vertebrate retina provide interesting examples of retinotopic spatial organization. However, it is not clear whether these two systems are spatially interrelated or not. The current work applies two mathematical-computational methods-a new method involving Voronoi diagrams for local density quantification and a more traditional approach, the Ripley K function-in order to characterize the mosaics of normally placed and displaced amacrine cells in the retina of Hoplias malabaricus and search for possible spatial relationships between these two types of mosaics. The results obtained by the Voronoi local density analysis suggest that the two systems of amacrine cells are spatially interrelated through nearly constant local density ratios. PMID- 17426354 TI - Web-based method for translating neurodevelopment from laboratory species to humans. AB - Biomedical researchers and medical professionals are regularly required to compare a vast quantity of neurodevelopmental literature obtained from an assortment of mammals whose brains grow at diverse rates, including fast developing experimental rodent species and slower developing humans. In this article, we introduce a database-driven website, which was created to address this problem using statistical-based algorithms to integrate hundreds of empirically derived developing neural events in 10 mammalian species (http://translatingtime.net/). The site, based on a statistical model that has evolved over the past decade, currently incorporates 102 different neurodevelopmental events obtained from 10 species: hamsters, mice, rats, rabbits, spiny mice, guinea pigs, ferrets, cats, rhesus monkeys, and humans. Data are arranged in a Structured Query Language database, which allows comparative brain development measured in postconception days to be converted and accessed in real time, using Hypertext Preprocessor language. Algorithms applied to the database also allow predictions for dates of specific neurodevelopmental events where empirical data are not available, including for the human embryo and fetus. By designing a web-based portal, we seek to make these comparative data readily available to all those who need to efficiently estimate the timing of neurodevelopmental events in the human fetus, laboratory species, or across several different species. In an effort to further refine and expand the applicability of this database, we include a mechanism to submit additional data. PMID- 17426355 TI - Cutting edge issues in allergy and clinical immunology. AB - Approximately every 5 yr, Clinical Reviews in Allergy and Immunology deviates from its usual practice of publishing volumes devoted to one theme to including papers that cover a range of subjects. This issue is one such exception and arose following a symposium at the International Institute for Research in Autoimmune Diseases named AESKU.KIPP Institute at their Wendelsheim facility. The AESKU.KIPP Institute was a particularly venue because it was initially established by a German diagnostic company and a Swiss benefactor, Karl- Heinz Kipp. The goal of the Institute was to develop a unique atmosphere to encourage original research in the field of autoimmunity and clinical immunology. The thought was to create an institute where young scientists from throughout the world could come for short periods of time to learn newer methodologies in both clinical immunology and also molecular biology. This theme contains several of the papers presented at the opening of the Institute and are incorporated herein because they focus on several unique aspects of clinical immunology, often referred to as the mosaic of autoimmunity. PMID- 17426356 TI - Hughes Syndrome: the antiphospholipid syndrome--a clinical overview. AB - In 1983, a detailed clinical description of a new syndrome was published. This prothrombotic syndrome was initially called the anticardiolipin syndrome and subsequently the antiphospholipid syndrome (APS), or Hughes Syndrome. Almost uniquely, it results in arterial as well as venous thrombosis and is marked by the presence of circulating antiphospholipid antibodies. Clinical features are protean, ranging from peripheral deep vein thrombosis (DVT) to involvement of internal organs such as the liver, kidneys, and adrenals. Likewise, arterial thrombosis can result in life-threatening infarction of organs such as the heart. The nervous system is frequently affected, with migraine, memory loss, balance disorders, stroke, and atypical multiple sclerosis being prominent. Other features include recurrent miscarriage, thrombocytopenia, and livedo reticularis. More recent observations have included ischemic bone fractures, renal and celiac artery stenosis, and a possible tendency toward accelerated atherosclerosis. The condition is seen in patients with lupus, but, significantly, occurs without associated lupus ("primary" APS)-indeed, increasing clinical recognition of Hughes Syndrome suggests that this condition will overtake lupus in prevalence. Treatment at present is by anticoagulation. The mechanisms for thrombosis are being worked out; it has been suggested that in some situations (e.g., pregnancy loss), an inflammatory component as well as thrombosis may play a part. PMID- 17426357 TI - The immune response to citrullinated proteins in patients with rheumatoid arthritis: genetic, clinical, technical, and epidemiological aspects. AB - This article reviews data concerning the applicability of anti-citrullinated peptide antibodies in the diagnosis, estimation of prognosis, and follow-up of patients with rheumatoid arthritis (RA). The production of anti-citrullinated peptide antibodies is closely associated with the presence of the HLA-DRB1 shared epitope, a known risk factor for development of RA, and the production may be influenced by environmental factors such as tobacco smoking. Patients who harbor this antibody from the early stage of their disease develop more severe erosive disease than patients with RA who lack the antibody. The anti-citrullinated peptide antibody level may be a reflection of disease activity, at least in the early phase of the disease. The antibody can sometimes be found several years before the onset of clinical symptoms of RA, which may represent an open window for preventive measures to be taken. PMID- 17426358 TI - Auto-antibodies and autoreactive T-cells in rheumatoid arthritis: pathogenetic players and diagnostic tools. AB - Rheumatoid arthritis (RA) is not only the most severe of all joint diseases but also the most common systemic autoimmune disease affecting approximately 1% of the world-wide adult population. RA is characterized by the presence of autoantibodies in serum and synovial fluid distinguishing the disease from other arthritides such reactive arthritis or osteoarthritis. Since the historical description of rheumatoid factor (RF), which is an autoantibody directed to immunoglobulin G, numerous additional autoantibodies have been discovered in sera of RA patients. These antibodies may be directed to cartilage components, stress proteins, enzymes, nuclear proteins and, most importantly, citrullinated proteins such as fibrin or vimentin. In contrast to other antibodies including RF, anti citrullinated protein antibodies are targeted almost exclusively by RA patients thus being the most specific serological markers of RA. Even though most other antibodies are not used for diagnostics, they may contribute to the patholophysiology of RA by forming immune complexes in the joint. Furthermore, autoreactive T-cells in serum and synovial fluid may initiate or enhance the disease process via production of proinflammatory cytokines leading to autoantibody secretion, stimulation of macrophages and activation of bone resorbing osteoclasts. Identification of novel autoantigens, particularly citrullinated proteins, and the characterization of the cellular and molecular processes underlying the autoimmune reactions against them has provided new insights into the complex pathogenesis of RA. This has made possible the development of novel therapeutic concepts that may allow to treat the disease more effectively in its early stages where the chances are highest to interrupt the deleterious processes. PMID- 17426359 TI - Are anti-ribosomal P protein antibodies relevant in systemic lupus erythematosus? AB - Systemic lupus erythematosus (SLE) is a prototypal auto-immune disorder characterized with multiple organ involvement resulting in disability and increased mortality. Immune regulatory disturbances cumulate in activation of B cells and consequent auto-antibody production. Antigens for these auto-antibodies can be nuclear components and cytoplasmic elements. Anti-P antibodies react against acidic phosphorylated ribosomal proteins P0, P1, and P2 (with molecular mass of 38, 19, and 17 kDa, respectively) and are located on the S60 subunit of ribosomes. Ribosomal P proteins share a common 22-amino acid sequence that is present in the carboxyl-terminal. Anti-P antibodies can be detected in approx 15 to 20% of patients with lupus by several immunoassays, most frequently by enzyme linked immunosorbent assay (ELISA) and/or Western blotting. However, no standardized assay is available. Auto-antibodies against eukaryotic P proteins appear highly specific for SLE; therefore, they can be used as diagnostic marker for the disease. Furthermore, association has been described with particular manifestations of lupus, especially with neuropsychiatric, renal, and hepatic involvements. Anti-P positivity and the titer of anti-P antibodies also fluctuate with clinical disease activity. Despite several lines of evidence, results are conflicting regarding the existence of such associations. Discrepancies can be explained by different study set-up or study population; it also can be attributed to the different sensitivity of tests used for the detection of anti-P antibody. PMID- 17426360 TI - Increased prevalence of anti-third generation cyclic citrullinated peptide antibodies in patients with rheumatoid arthritis and CREST syndrome. AB - To investigate the prevalence of anti-third generation cyclic citrullinated peptide antibodies (anti-CCP3) in patients with systemic connective tissue diseases, we assembled a training set consisting of 115 patients with rheumatoid arthritis (RA), 52 with Calcinosis, Raynaud's phenomenon, oesophageal dysmotility, sclerodactyly, telangiectasia (CREST) syndrome, 21 with scleroderma, 20 with ankylosing spondylitis, 18 with reactive arthritis, 25 with juvenile rheumatoid arthritis (RA), 51 with osteoarthritis, 26 with mixed connective tissue disease, 23 with primary Sjogren's syndrome, 74 with systemic lupus erythematosus, 49 with Polymyalgia rheumatica, and 39 with polymyositis/dermatomyositis. The commercial enzyme-linked immunosorbent assay (ELISA) was used to detect anti-CCP antibodies, including anti-CCP2 (regular, second generation of CCP antigen) and anti-CCP3 (third generation of CCP antigen) in disease-related specimens and normal controls. These serum samples were also evaluated for anti-centomere antibodies by anti-centromere ELISA kit. The higher frequencies of anti-CCP3 and anti-CCP2 were detected in 75.6 and 70.4% patients with RA, respectively. At the same time, anti-CCP3 (not anti-CCP2) was significantly increased in samples isolated from patients with CREST syndrome. The clinical sensitivity of IgG anti-CCP3 for the patients with CREST syndrome was 29% (15 of 52) and the specificity was 96% (384 of 397), with the exception of the RA group. The anti-centromere antibodies were significantly higher in patients with CREST only. The results of our study suggest that compared to anti CCP2 assay, the new anti-CCP3 assay can enhance the clinical sensitivity for diagnosis of RA and, as an associate marker combined with anticentromere, can distinguish CREST syndrome from other systemic connective tissue diseases, especially RA. The clinical specificity of anti-CCP3 was lower than anti-CCP2 assay in diagnosis of RA because of the crossreaction to the patients with CREST syndrome. PMID- 17426361 TI - Transcutaneous immunization with imiquimod is amplified by CD40 ligation and results in sustained cytotoxic T-lymphocyte activation and tumor protection. AB - Transcutaneous immunization (TCI) using ligands of Toll-like receptors (TLRs) and cytotoxic T-lymphocyte (CTL) epitopes lead to the induction of potent T-cell responses. To characterize the efficacy of TCI-mediated CTL activation, we monitored the frequency and functional activity of specific CTL induced with TCI using the ovalbumin-derived epitope SIINFEKL composed in creme containing the synthetic TLR7 ligand R-837. We found that the frequency and activity decayed rapidly 10 d post-TCI. Consistently, no significant memory T-cell formation was detectable. In a prophylactic vaccination setting, TCI was protective against a lethal challenge with ovalbumin expressing EG.7 thymoma cells when the tumor cells were inoculated 5 d later. However, only a delay of tumor growth was observed when the tumor challenge was performed 55 d after immunization. Conversely, a single combined treatment with TCI and an agonist anti-CD40 (FGK 45) monoclonal antibody greatly enhanced the primary response, with up to 30% of peptide-specific CTL and the effective induction of memory cells. Consequently, mice treated with TCI/anti-CD40 were completely protected against a lethal tumor challenge with EG.7 tumor cells after 55 d. In this article, we demonstrate that transcutaneous immunization approaches using TLR ligands deliver sufficient amounts of antigen to mediate durable protection against tumors if adequate costimulation is provided. These results may contribute to the development of advanced vaccination protocols against malignancies and persistent virus infections. PMID- 17426362 TI - Role of anti-beta2 glycoprotein I antibodies in antiphospholipid syndrome: in vitro and in vivo studies. AB - Antiphospholipid syndrome (APS) is characterized by the presence of recurrent venous/ arterial thrombosis and fetal losses associated with a family of auto antibodies directed against phospholipid (PL)-binding proteins. Among them, beta2 glycoprotein I (beta2GPI) is the most important. As a plasma cationic protein, beta2GPI binds to anionic PLs involved in several fluid-phase coagulation steps, and more importantly, it can be expressed on the surface of different cell types. Anti-beta2GPI antibodies recognize the molecule expressed on endothelial cells, platelets, monocytes, and trophoblast cells. Once bound, the antibodies trigger in vitro cell signaling that modulates biological responses potentially responsible for pathogenic mechanisms. Experimental animal models have supported the in vivo pathogenic role of anti-beta2GPI antibodies in both thrombosis and fetal loss models. PMID- 17426363 TI - High-content T-cell and B-cell analysis. AB - Immune responses are orchestrated by B- and T-cells endowed with a diverse repertoire of antigen receptors. Comprehensive profiling of the humoral and cellular immune responses, including molecular composition, the specificity of the response, the cellular activation, differentiation, and homing status, will aid to improve diagnosis, classification, monitoring, and prognostication of immune disorders driven by antigen-specific B- and T-lymphocytes. PMID- 17426364 TI - Indications for biotherapy in systemic vasculitides. AB - Biotherapy now holds a specific place in the therapeutic armamentarium for systemic vasculitides. Such therapy includes cytokines, such as (pegylated) alpha interferon for hepatitis B virus-related polyarteritis nodosa and hepatitis C virus-related cryoglobulinemic vasculitis, and polyvalent immunoglobulin (IVIg), with well-defined indications and pending positive results. More specifically targeted monoclonal antibodies include antitumor necrosis factor-alpha or anti CD20 for antineutrophil cytoplasmic antibody-associated vasculitides or anti interleukin-5 and anti-IgE for Churg-Strauss syndrome. However, the exact indications of these latter new agents, as well as their optimal dosage and duration, are not defined. Therefore, they are prescribed mainly for patients with disease refractory to conventional therapy, in whom results are promising. Results of international ongoing trials will determine whether the agents may also have a place as first-line treatment. PMID- 17426365 TI - Aspirin and nonsteroidal anti-inflammatory drug hypersensitivity. AB - Acetylsalicylic acid (ASA) or aspirin and nonsteroidal anti-inflammatory drug (NSAID) sensitivities encompass a diverse group of both pharmacological and hypersensitivity reactions. Conventionally, hypersensitivities include aspirin exacerbated respiratory disease (AERD), ASA-induced urticaria, and anaphylaxis. With an increasing prevalence of coronary artery disease in an aging population, aspirin continues to play a significant role in cardiac prophylaxis in a large patient population. Invariably, the clinician will encounter patients with clear indications for aspirin therapy but a history of aspirin sensitivity. Although protocols have been established for aspirin challenge and desensitization, it is not always an efficacious or safe procedure. This article reviews the different classifications of ASA/NSAIDs hypersensitivities to better guide the clinician in dealing with this patient population. History of crossrelativities between multiple NSAIDs implies a non-IgE-mediated process. Similarly, a history of monosensitivity to one NSAID implies an IgE-mediated process, although specific antibodies are often elusive. Despite the name, AERD can potentially be exacerbated by all cyclooxygenase (COX) inhibitors based on dose-dependent inhibition of COX-1. Aspirin desensitization can be achieved to improve both upper and lower respiratory symptoms for most patient with AERD. Aspirin desensitization can usually be achieved for those in need of the antiplatelet effects of aspirin, with the exception of those with aspirin-induced urticaria and baseline chronic urticaria. However, desensitization should only be attempted in those with stable coronary artery disease because the process of desensitization carries the inherent risk of anaphylaxis/anaphylactoid reaction, which may further increase cardiac demand and bring about ischemic injury. Therefore, desensitization is reserved until coronary artery disease is stabilized. PMID- 17426366 TI - Molecular mimicry and auto-immunity. AB - The term "molecular mimicry" was coined by R. Damian in 1964, who was first to suggest that antigenic determinants of micro-organisms may resemble antigenic determinants of their host. Damian suggested that this similarity served as a defense mechanism of a microorganism from the host's immune system and prevented the development of immune response to the micro-organism, thereby protecting it from host defense. Years later, the term "molecular mimicry" was attributed a different meaning-namely, antigenic determinants of microorganisms might elicit an auto-immune response that harms the host. The concept of molecular mimicry is based on a structural similarity between a pathogen or metabolite and self structures. The similarity could be expressed as shared amino acid sequences (linear or mimotope) or similar conformational structure between a pathogen and self-antigen. "Molecular mimicry" has become a very popular explanation for the frequent association of infection with auto-immune disease. PMID- 17426367 TI - A critical review of local anesthetic sensitivity. AB - With their ability to block pain signals to the brain, local anesthetics (LAs) have made possible many surgical procedures and interventions once thought impossible. LAs are generally safe and well tolerated when used correctly by trained professionals. However, adverse reactions do occur, and may generate a referral to an Allergist for evaluation of LA allergy. LA structure, classification, and metabolism will be briefly reviewed. A critical analysis of the studies and case reports involving LA allergy found via PubMed search for "local anesthetic allergy" and "local anesthetic hypersensitivity" will be discussed. In addition, the clinical evaluation of a patient with concern for a LA allergy will be examined. PMID- 17426368 TI - Injecting drug use in pregnant HIV-infected women in Europe. AB - OBJECTIVE: to describe the characteristics of pregnant HIV-infected women with past or current injecting drug use (IDU) and their infants in a large European cohort. DESIGN: prospective study. METHODS: analysis of data on HIV-infected pregnant women enrolling in the European Collaborative Study, 1985 to May 2005. Factors examined included injecting behaviour, HIV-related and socio-demographic characteristics and pregnancy outcomes. RESULTS: of the 5566 women enrolled, 2342 (42%) reported ever using injecting drugs, including 862 current users. Most IDUs were white and nulliparous at enrollment. Current IDUs were younger than ex-IDUs (respective median ages, 26.7 and 28.1 years), less likely to be married or cohabiting (57% (319/559) versus 74% (601/809), p<0.001) and more likely to report an IDU sexual partner (63% (483/762) versus 42% (548/1307) p<0.001). Of the current IDUs, 768 (89%) used illicit drugs throughout their pregnancy, with 567 having an infant with drug withdrawal symptoms. Overall, 42% (399/944) IDUs had a CD4 count of >or=500 cells/mm3, with current IDUs slightly more at risk of severe immunosuppression than ex-IDUs (17% versus 12%, p=0.058). Of the 2515 mother-child pairs, 1716 (68%) received no antenatal antiretroviral prophylaxis, 503 (20%) monotherapy or dual therapy and 296 (12%) HAART. The MTCT rate among women receiving any ART was 4.06% (95% CI 2.62-5.98) and 0.93% (95% CI 0.11-3.34) among those on HAART. In multivariable analysis, elective caesarean section was associated with an 82% reduced MTCT risk versus vaginal delivery (AOR 0.18, p<0.001) and maternal HAART with a 77% reduced risk versus no antenatal ART (AOR 0.23, p<0.001). CONCLUSION: among the HIV-infected pregnant women in this cohort, characteristics of current and ex-IDUs differed considerably. However, MTCT risk did not vary according to IDU, and risk factors for transmission were as described for women with no history of IDU in this cohort and elsewhere. PMID- 17426369 TI - [Acid-base balance and umbilical blood gases in neonates born at term in North Eastern region of Poland]. AB - INTRODUCTION: acidosis observed in infants in the neonatal period can be the cause of many disturbances in adaptation to extra uterine life. In the literature various values of acid-base balance and umbilical blood gases parameters are quoted as borderline between normal and pathological. AIM: of study was to determine the parameters of acid-base balance and umbilical venous and arterial blood gases in newborn infants from North-Eastern Poland, significant for the evaluation of the condition of the neonate population from this region. MATERIAL AND METHODS: three groups of newborns were selected for the study. Group I consisted of 492 neonates, born at term, by spontaneous vaginal birth, from single pregnancies, without detectable developmental malformations, without traits of hypotrophy or macrosomy, with normal record of foetal cardiac activity during pregnancy and delivery. Apgar score at 5 min. was >7. Group II consisted of 121 infants born by Caesarean section due to maternal indications. Group III, (75 neonates) was selected from infants born at term by vaginal birth, with Apgar score 422 ng/ml allows confirmation of intrauterine origin of infection symptoms in pregnant women and allows the prediction of infection in infants born of women who showed symptoms of infection. PMID- 17426371 TI - [Concentration of procalcitonin in maternal blood, amniotic fluid and in umbilical cord blood as an indicator of neonatal infection]. AB - THE AIM of this study was to evaluate the usefulness of determining the concentration of procalcitonin (PCT) in the blood of pregnant women, in amniotic fluid and in umbilical blood as a predictor of intrauterine or infant infection. MATERIAL AND METHODS: the study was conducted on the following material: pregnant women s blood, amniotic fluid and umbilical blood taken during delivery from 60 women and 60 infants. In this group there were 30 vaginal deliveries and 30 deliveries by caesarean section. The PCT concentration was determined by an immunoluminometrical method (ILMA) using two antigen specific monoclonal antibodies; method sensitivity was 0.3 ng/ml. The determining precision was 5 10%. In statistical analysis the U Mann- Whitney test, chi square and ROC curve were employed. A p value less then 0.05 was considered as statistically significant. RESULTS: statistically significant increase in concentration of PCT (p<0.049) was found in umbilical blood of infants whose mothers had intrauterine infection, while such a correlation was not found either in the pregnant women's blood nor in the amniotic fluid. The low levels of AUC indicators found in all analyzed systemic fluids indicate minor usefulness of PCT as an indicator allowing predictions of infant infections. PCT concentrations in the amniotic fluid in comparison with concentrations of PCT in pregnant women's blood and in umbilical blood showed the highest value as a predictor of infection in infants after delivery (respectively AUC - 0.599; 0.582; 0.545). However, low levels of indicator LR - 1.35 demonstrate a nonsignificant value of this indicator as a predictor of infection in infants independent of infection in pregnant women, infection in pregnant women and their infants, or premature delivery. CONCLUSIONS: 1. Clinical symptoms of infection in pregnant women are accompanied by increased concentration of PCT in the umbilical blood. 2. The absence of symptoms of infection in a pregnant woman, infection in infant or simultaneous infection in infant and pregnant woman is not a predictor of concentrations of PCT in pregnant woman blood, in amniotic fluid nor in umbilical blood. 3. Examining the concentration of PCT in blood of pregnant women, in amniotic fluid and in umbilical blood is not a clinically useful indicator predicting intrauterine infection in infant. PMID- 17426372 TI - [Newborn life threatening respiratory failure treatment with extracorporeal membrane oxygenation]. AB - THE AIM of the study was to show first results of newborn life threatening respiratory failure treatment with extracorporeal membrane oxygenation (ECMO) in Poland. MATERIAL: Nine newborns were treated with extracorporeal membrane oxygenation in Silesian Center for Heart Diseases. Newborns were born in 38 week of gestational age (36-41 weeks) with mean birth weight of 3490 g. Reasons for the referral were: meconium aspiration syndrome, infection, and pulmonary hypertension. Each newborn fulfilled an Extracorporeal Life Support Organization (ELSO) criteria for extracorporeal membrane oxygenation. RESULTS: seven out of nine of patients treated with extracorporeal membrane oxygenation survived. Full clinical stabilization was reached about 6th hour of treatment. Mean extracorporeal oxygenation time was 162 hours. For eight newborns veno-venous method was applied and for one newborn veno-arterial method. Roller pump was used in 7 cases and centrifugal pomp in one case. Five newborns had uneventful treatment. During extracorporeal membrane oxygenation therapy we have observed several complications: PDA, hemorrhagic complications, renal failure, arterial hypertension, septicemia, tubing rupture. CONCLUSIONS: extracorporeal oxygenation is an effective method of treatment for newborn life threatening respiratory failure. Obtained results do not differ much from Extracorporeal Life Support Organization register results. The most essential problem for extracorporeal membrane oxygenation therapy is correct qualification, early referral, safe transportation as well as the development of centers providing ECMO treatment. PMID- 17426373 TI - [Chorioamnionitis in relation to cord blood sICAM-1 and CRP levels of newborns with and without markers of early onset infection]. AB - THE AIM of the study was an evaluation of the relationship between histological chorioamnionitis and cord blood soluble intercellular adhesion molecule-1 (sICAM 1) and CRP levels. MATERIAL AND METHODS: in the group of 152 neonates observed for early onset infection (clinical and laboratory signs) the relationship between histological chorioamnionitis and cord blood sICAM-1 (ELISA) and CRP (turbidimetric method) levels was assessed. All these newborns were born in the years 2002-2004 to mothers with risk factors of perinatal infection. RESULTS: histological chorioamnionitis (ChA) was found in 50 cases, while early onset infection signs occurred in 112 neonates. The percentage of ill and healthy newborns in group with ChA and without ChA was similar. There were significantly higher sICAM-1 (p<0.001) levels in neonates from pregnancy complicated with ChA than without ChA (179 +/- 68.7 v. 147 +/- 30.8 ng/ml). There was not such difference in the CRP levels. There was also a significant positive correlation between cord blood sICAM-1 and ChA in the whole analyzed group of patients (r=0.26; p<0.001) and in the group of ill neonates (r=0.37; p<0.0001). There was a significant difference (p<0.001) in sICAM-1 levels between ill and healthy neonates from the group with ChA (191,8 +/- 73.9 v. 143,4 +/- 31.9 ng/ml) but not without ChA. No difference was found in sICAM-1 levels between groups of healthy neonates born after pregnancy complicated and not complicated with ChA. CONCLUSIONS: coexistence of ChA and early onset infection in neonates enhanced the cord blood sICAM-1 levels. Healthy newborns born from pregnancy complicated by ChA do not react by increasing cord blood sICAM-1 levels. It is not possible to exclude early onset infection in neonates on the basis of normal cord blood sICAM-1 levels, because ill newborns born after pregnancy not complicated with ChA also do not react with increasing sICAM-1 production at birth. PMID- 17426374 TI - [Late form of vitamin K deficiency bleeding. Description of four cases with various clinical picture and outcome]. AB - Late form of vitamin K deficiency bleeding (VKDB) occurs in infants who received none, or inadequate vitamin K prophylaxis after birth. Exclusively breastfed infants are especially prone to development of this disease. We present 4 cases of late VKDB with different clinical picture and outcome. In two patients intracranial bleeding of variable magnitude was detected, in one, bleeding into pleural cavity and in the fourth severe bleeding from injection sites without internal haemorrhage. Diagnosis was based on the clinical picture with special emphasis on the child's past history and results of coagulation tests. Treatment consisted mainly of vitamin K administration and infusion of fresh frozen plasma. One patient developed severe and definite brain damage after intracranial bleeding, the second only a mild brain dysfunction, the remaining two infants recovered fully. These observations and literature data suggest that in exclusively breastfed infants who did not receive vitamin K after birth, late VKDB should be taken into account as a cause of haemorrhage with different localization and magnitude. There is a need for more emphasis on teaching of physicians about late VKDB and for parenteral vitamin K prophylaxis programme for all newborn infants. PMID- 17426375 TI - [Alveolar capillary dysplasia as a cause of failure in treatment of a neonate with pulmonary persistent hypertension of the newborn - case report]. AB - A patient with severe persistent pulmonary hypertension of the newborn (PPHN) due to alveolar capillary dysplasia, congenital (ACD), is presented. In the treatment, apart from standard methods, high frequency oscillatory ventilation (HFOV), inhaled nitric oxide and activated C protein have been applied. In spite of treatment the patient died and post-mortem diagnosis was based on lung histopathology examination. ACD occurs very rarely and is a congenital disease. Diagnosis is by pulmonary tissue histopathology examination. Pathological structure of the lungs leads to severe dysfunction of gas exchange as well as increasing pulmonary hypertension. No effective treatment is known and all so far described cases have ended up with death. The described case and literature data lead the authors to the following conclusions: 1. in case of PPHN resistant to treatment, ACD diagnosis should be taken into consideration, 2. histopathological examination determines the diagnosis, 3. limited capabilities of diagnosis are the reason for applying non-standard and expensive treatment methods which so far are doomed to failure, 4. in case of a patient with severe, persistent pulmonary hypertension and unclear aetiology, not reacting to nitrous oxide treatment, a diagnostic lung biopsy should be considered. PMID- 17426376 TI - Simple sequence repeats in mycobacterial genomes. AB - Simple sequence repeats (SSRs) or microsatellites are the repetitive nucleotide sequences of motifs of length 1-6 bp. They are scattered throughout the genomes of all the known organisms ranging from viruses to eukaryotes. Microsatellites undergo mutations in the form of insertions and deletions (INDELS) of their repeat units with some bias towards insertions that lead to microsatellite tract expansion. Although prokaryotic genomes derive some plasticity due to microsatellite mutations they have in-built mechanisms to arrest undue expansions of microsatellites and one such mechanism is constituted by post-replicative DNA repair enzymes MutL, MutH and MutS. The mycobacterial genomes lack these enzymes and as a null hypothesis one could expect these genomes to harbour many long tracts. It is therefore interesting to analyse the mycobacterial genomes for distribution and abundance of microsatellites tracts and to look for potentially polymorphic microsatellites. Available mycobacterial genomes, Mycobacterium avium, M. leprae, M. bovis and the two strains of M. tuberculosis (CDC1551 and H37Rv) were analysed for frequencies and abundance of SSRs. Our analysis revealed that the SSRs are distributed throughout the mycobacterial genomes at an average of 220-230 SSR tracts per kb. All the mycobacterial genomes contain few regions that are conspicuously denser or poorer in microsatellites compared to their expected genome averages. The genomes distinctly show scarcity of long microsatellites despite the absence of a post-replicative DNA repair system. Such severe scarcity of long microsatellites could arise as a result of strong selection pressures operating against long and unstable sequences although influence of GC-content and role of point mutations in arresting microsatellite expansions can not be ruled out. Nonetheless, the long tracts occasionally found in coding as well as non-coding regions may account for limited genome plasticity in these genomes. PMID- 17426377 TI - Sequence and recombination analyses of the geminivirus replication initiator protein. AB - The sequence motifs present in the replication initiator protein (Rep) of geminiviruses have been compared with those present in all known rolling circle replication initiators. The predicted secondary structures of Rep representing each group of organisms have been compared and found to be conserved. Regions of recombination in the Rep gene and the adjoining 5' intergenic region (IR)of representative species of Geminiviridae have been identified using Recombination Detection Programs. The possible implications of such recombinations on the increasing host range of geminivirus infections are discussed. PMID- 17426378 TI - A hybrid approach for predicting promiscuous MHC class I restricted T cell epitopes. AB - In the present study, a systematic attempt has been made to develop an accurate method for predicting MHC class I restricted T cell epitopes for a large number of MHC class I alleles. Initially, a quantitative matrix (QM)-based method was developed for 47 MHC class I alleles having at least 15 binders. A secondary artificial neural network (ANN)-based method was developed for 30 out of 47 MHC alleles having a minimum of 40 binders. Combination of these ANN-and QM-based prediction methods for 30 alleles improved the accuracy of prediction by 6% compared to each individual method. Average accuracy of hybrid method for 30 MHC alleles is 92.8%. This method also allows prediction of binders for 20 additional alleles using QM that has been reported in the literature, thus allowing prediction for 67 MHC class I alleles. The performance of the method was evaluated using jack-knife validation test. The performance of the methods was also evaluated on blind or independent data. Comparison of our method with existing MHC binder prediction methods for alleles studied by both methods shows that our method is superior to other existing methods. This method also identifies proteasomal cleavage sites in antigen sequences by implementing the matrices described earlier. Thus, the method that we discover allows the identification of MHC class I binders (peptides binding with many MHC alleles) having proteasomal cleavage site at C-terminus. The user-friendly result display format (HTML-II) can assist in locating the promiscuous MHC binding regions from antigen sequence. The method is available on the web at www.imtech.res.in/raghava/nhlapred and its mirror site is available at http://bioinformatics.uams.edu/mirror/nhlapred/. PMID- 17426379 TI - Genome inventory and analysis of nuclear hormone receptors in Tetraodon nigroviridis. AB - Nuclear hormone receptors (NRs) form a large superfamily of ligand-activated transcription factors, which regulate genes underlying a wide range of (patho) physiological phenomena. Availability of the full genome sequence of Tetraodon nigroviridis facilitated a genome wide analysis of the NRs in fish genome. Seventy one NRs were found in Tetraodon and were compared with mammalian and fish NR family members. In general, there is a higher representation of NRs in fish genomes compared to mammalian ones. They showed high diversity across classes as observed by phylogenetic analysis. Nucleotide substitution rates show strong negative selection among fish NRs except for pregnane x receptor (PxR), estrogen receptor (ER) and liver x receptor (LxR). This may be attributed to crucial role played by them in metabolism and detoxification of xenobiotic and endobiotic compounds and might have resulted in slight positive selection. Chromosomal mapping and pairwise comparisons of NR distribution in Tetraodon and humans led to the identification of nine syntenic NR regions, of which three are common among fully sequenced vertebrate genomes. Gene structure analysis shows strong conservation of exon structures among orthologoues. Whereas paralogous members show different splicing patterns with intron gain or loss and addition or substitution of exons played a major role in evolution of NR superfamily. PMID- 17426380 TI - "Pinning strategy": a novel approach for predicting the backbone structure in terms of protein blocks from sequence. AB - The description of protein 3D structures can be performed through a library of 3D fragments, named a structural alphabet. Our structural alphabet is composed of 16 small protein fragments of 5 C alpha in length, called protein blocks (PBs). It allows an efficient approximation of the 3D protein structures and a correct prediction of the local structure. The 72 most frequent series of 5 consecutive PBs, called structural words (SWs)are able to cover more than 90% of the 3D structures. PBs are highly conditioned by the presence of a limited number of transitions between them. In this study, we propose a new method called "pinning strategy" that used this specific feature to predict long protein fragments. Its goal is to define highly probable successions of PBs. It starts from the most probable SW and is then extended with overlapping SWs. Starting from an initial prediction rate of 34.4%, the use of the SWs instead of the PBs allows a gain of 4.5%. The pinning strategy simply applied to the SWs increases the prediction accuracy to 39.9%. In a second step, the sequence-structure relationship is optimized, the prediction accuracy reaches 43.6%. PMID- 17426381 TI - ProRegIn: a regularity index for the selection of native-like tertiary structures of proteins. AB - Automated protein tertiary structure prediction from sequence information alone remains an elusive goal to computational prescriptions. Dividing the problem into three stages viz. secondary structure prediction, generation of plausible main chain loop dihedrals and side chain dihedral optimization, considerable progress has been achieved in our laboratory (http://www.scfbio iitd.res.in/bhageerath/index.jsp) and elsewhere for proteins with less than 100 amino acids. As a part of our on-going efforts in this direction and to facilitate tertiary structure selection/rejection in containing the combinatorial explosion of trial structures for a specified amino acid sequence, we describe here a web-enabled tool ProRegIn (Protein Regularity Index) developed based on the regularity in the Phi, Psi dihedral angles of the amino acids that constitute loop regions. We have analysed the dihedrals in loop regions in a non-redundant dataset of 7351 proteins drawn from the Protein Data Bank and categorized them as helix-like or sheet-like (regular) or irregular. We noticed that the regularity thus defined exceeds 86% for Phi barring glycine and 70% for Psi for all the amino acid side chains including glycine, compelling us to reexamine the conventional view that loops are irregular regions structurally. The regularity index is presented here as a simple tool that finds its application in protein structure analysis as a discriminatory scoring function for rapid screening before the more compute intensive atomic level energy calculations could be undertaken. The tool is made freely accessible over the internet at www.scfbio iitd.res.in/software/proregin.jsp. PMID- 17426382 TI - Comparison of sequence-based and structure-based phylogenetic trees of homologous proteins: Inferences on protein evolution. AB - Several studies based on the known three-dimensional (3-D) structures of proteins show that two homologous proteins with insignificant sequence similarity could adopt a common fold and may perform same or similar biochemical functions. Hence, it is appropriate to use similarities in 3-D structure of proteins rather than the amino acid sequence similarities in modelling evolution of distantly related proteins. Here we present an assessment of using 3-D structures in modelling evolution of homologous proteins. Using a dataset of 108 protein domain families of known structures with at least 10 members per family we present a comparison of extent of structural and sequence dissimilarities among pairs of proteins which are inputs into the construction of phylogenetic trees. We find that correlation between the structure-based dissimilarity measures and the sequence based dissimilarity measures is usually good if the sequence similarity among the homologues is about 30% or more. For protein families with low sequence similarity among the members, the correlation coefficient between the sequence based and the structure-based dissimilarities are poor. In these cases the structure-based dendrogram clusters proteins with most similar biochemical functional properties better than the sequence-similarity based dendrogram. In multi-domain protein families and disulphide-rich protein families the correlation coefficient for the match of sequence-based and structure-based dissimilarity (SDM) measures can be poor though the sequence identity could be higher than 30%. Hence it is suggested that protein evolution is best modelled using 3-D structures if the sequence similarities (SSM) of the homologues are very low. PMID- 17426383 TI - SURF'S UP! - protein classification by surface comparisons. AB - Large-scale genome sequencing and structural genomics projects generate numerous sequences and structures for 'hypothetical' proteins without functional characterizations. Detection of homology to experimentally characterized proteins can provide functional clues, but the accuracy of homology-based predictions is limited by the paucity of tools for quantitative comparison of diverging residues responsible for the functional divergence. SURF'S UP! is a web server for analysis of functional relationships in protein families, as inferred from protein surface maps comparison according to the algorithm. It assigns a numerical score to the similarity between patterns of physicochemical features(charge, hydrophobicity) on compared protein surfaces. It allows recognizing clusters of proteins that have similar surfaces, hence presumably similar functions. The server takes as an input a set of protein coordinates and returns files with "spherical coordinates" of proteins in a PDB format and their graphical presentation, a matrix with values of mutual similarities between the surfaces, and the unrooted tree that represents the clustering of similar surfaces, calculated by the neighbor-joining method. SURF'S UP! facilitates the comparative analysis of physicochemical features of the surface, which are the key determinants of the protein function. By concentrating on coarse surface features, SURF'S UP! can work with models obtained from comparative modelling. Although it is designed to analyse the conservation among homologs, it can also be used to compare surfaces of non-homologous proteins with different three dimensional folds, as long as a functionally meaningful structural superposition is supplied by the user. Another valuable characteristic of our method is the lack of initial assumptions about the functional features to be compared. SURF'S UP! is freely available for academic researchers at http://asia.genesilico.pl/surfs_up/. PMID- 17426384 TI - Peptide segments in protein-protein interfaces. AB - An important component of functional genomics involves the understanding of protein association. The interfaces resulting from protein-protein interactions - (i) specific, as represented by the homodimeric quaternary structures and the complexes formed by two independently occurring protein components, and (ii) non specific, as observed in the crystal lattice of monomeric proteins - have been analysed on the basis of the length and the number of peptide segments. In 1000 A2 of the interface area, contributed by a polypeptide chain, there would be 3.4 segments in homodimers, 5.6 in complexes and 6.3 in crystal contacts. Concomitantly, the segments are the longest (with 8.7 interface residues) in homodimers. Core segments (likely to contribute more towards binding) are more in number in homodimers (1.7) than in crystal contacts (0.5), and this number can be used as one of the parameters to distinguish between the two types of interfaces. Dominant segments involved in specific interactions, along with their secondary structural features, are enumerated. PMID- 17426385 TI - Modelling and simulation of signal transductions in an apoptosis pathway by using timed Petri nets. AB - This paper first presents basic Petri net components representing molecular interactions and mechanisms of signalling pathways, and introduces a method to construct a Petri net model of a signalling pathway with these components. Then a simulation method of determining the delay time of transitions, by using timed Petri nets - i.e. the time taken in fi ring of each transition - is proposed based on some simple principles that the number of tokens flowed into a place is equivalent to the number of tokens fl owed out. Finally, the availability of proposed method is confirmed by observing signalling transductions in biological pathways through simulation experiments of the apoptosis signalling pathways as an example. PMID- 17426386 TI - Dynamic causal models of neural system dynamics:current state and future extensions. AB - Complex processes resulting from interaction of multiple elements can rarely be understood by analytical scientific approaches alone; additional, mathematical models of system dynamics are required. This insight, which disciplines like physics have embraced for a long time already, is gradually gaining importance in the study of cognitive processes by functional neuroimaging. In this field, causal mechanisms in neural systems are described in terms of effective connectivity. Recently, dynamic causal modelling (DCM) was introduced as a generic method to estimate effective connectivity from neuroimaging data in a Bayesian fashion. One of the key advantages of DCM over previous methods is that it distinguishes between neural state equations and modality-specific forward models that translate neural activity into a measured signal. Another strength is its natural relation to Bayesian model selection (BMS) procedures. In this article, we review the conceptual and mathematical basis of DCM and its implementation for functional magnetic resonance imaging data and event-related potentials. After introducing the application of BMS in the context of DCM, we conclude with an outlook to future extensions of DCM. These extensions are guided by the long-term goal of using dynamic system models for pharmacological and clinical applications, particularly with regard to synaptic plasticity. PMID- 17426387 TI - Formal TCA cycle description based on elementary actions. AB - Many databases propose their own structure and format to provide data describing biological processes. This heterogeneity contributes to the difficulty of large systematic and automatic functional comparisons. To overcome these problems, we have used the BioPsi formal description scheme which allows multi-level representations of biological process information. Applied to the description of the tricarboxylic acid cycle (TCA), we show that BioPsi allows the formal integration of functional information existing in current databases and make them available for further automated analysis. In addition such a formal TCA cycle process description leads to a more accurate biological process annotation which takes in account the biological context. This enables us to perform an automated comparison of the TCA cycles for seven different species based on processes rather than protein sequences. From current databases, BioPsi is able to unravel information that are already known by the biologists but are not available for automated analysis tools and simulation software, because of the lack of formal process descriptions. This use of the BioPsi description scheme to describe the TCA cycle was a key step of the MitoScop project that aims to describe and simulate mitochondrial metabolism in silico. PMID- 17426388 TI - Modelling spatio-temporal interactions within the cell. AB - Biological phenomena at the cellular level can be represented by various types of mathematical formulations. Such representations allow us to carry out numerical simulations that provide mechanistic insights into complex behaviours of biological systems and also generate hypotheses that can be experimentally tested. Currently, we are particularly interested in spatio-temporal representations of dynamic cellular phenomena and how such models can be used to understand biological specificity in functional responses. This review describes the capability and limitations of the approaches used to study spatio-temporal dynamics of cell signalling components. PMID- 17426389 TI - Integrative content-driven concepts for bioinformatics "beyond the cell". AB - Bioinformatics has delivered great contributions to genome and genomics research, without which the world-wide success of this and other global ('omics') approaches would not have been possible. More recently, it has developed further towards the analysis of different kinds of networks thus laying the foundation for comprehensive description, analysis and manipulation of whole living systems in modern "systems biology". The next step which is necessary for developing a systems biology that deals with systemic phenomena is to expand the existing and develop new methodologies that are appropriate to characterize intercellular processes and interactions without omitting the causal underlying molecular mechanisms. Modelling the processes on the different levels of complexity involved requires a comprehensive integration of information on gene regulatory events, signal transduction pathways, protein interaction and metabolic networks as well as cellular functions in the respective tissues / organs. PMID- 17426390 TI - Transient receptor potential channel TRPM8 agonists stimulate calcium influx and neurotensin secretion in neuroendocrine tumor cells. AB - TRPM8 is a member of the melastatin-type transient receptor potential ion channel family. Activation by cold or by agonists (menthol, icilin) induces a transient rise in intracellular free calcium concentration ([Ca(2+)](i)). Our previous study demonstrated that Ca(2+)-permeable cation channels play a role in IGF-1 induced secretion of chromogranin A in human neuroendocrine tumor (NET) cell line BON [Mergler et al.: Neuroendocrinology 2006;82:87-102]. Here, we extend our earlier study by investigating the expression of TRPM8 and characterizing its impact on [Ca(2+)](i) and the secretion of neurotensin (NT). We identified TRPM8 expression in NET BON cells by RT-PCR, Western blotting and immunofluorescence staining. Icilin increased [Ca(2+)](i) in TRPM8-transfected human embryonic kidney cells (HEK293) but not in mock-transfected cells. Icilin and menthol induced Ca(2+) transients in BON cells as well as in primary NET cell cultures of two different pancreatic NETs as detected by single cell fluorescence imaging. Icilin increased non-selective cation channel currents in BON cells as detected by patch-clamp recordings. This activation was associated with increased NT secretion. Taken together, this study demonstrates for the first time the expression TRPM8 in NET cells and its role in regulating [Ca(2+)](i) and NT secretion. The regulation of NT secretion in NETs by TRPM8 may have a potential clinical implication in diagnosis or therapy. PMID- 17426391 TI - Corticosterone impairs insulin-stimulated translocation of GLUT4 in the rat hippocampus. AB - BACKGROUND: Exposure to stress levels of glucocorticoids produces physiological responses that are characteristic of type 2 diabetes, such as peripheral insulin resistance and impairment in insulin-stimulated trafficking of glucose transporter 4 (GLUT4) in muscle and fat. In the central nervous system, stress produces neuroanatomical and neurochemical changes in the hippocampus that are associated with cognitive impairments. METHODS: In view of these observations, the current studies examined the effects of short-term (1 week) exposure of stress levels of glucocorticoids upon insulin receptor (IR) expression and signaling, including GLUT4 translocation, in the rat hippocampus. RESULTS: One week of corticosterone (CORT) treatment produced insulin resistance in response to peripheral glucose challenge. In the hippocampus, IR expression was unchanged in CORT-treated rats as compared with vehicle-treated rats. However, insulin stimulated phosphorylation of the IR, total Akt levels and total GLUT4 levels were reduced in CORT-treated rats when compared to controls. In addition, insulin stimulated translocation of hippocampal GLUT4 to the plasma membrane was completely abolished in CORT-treated rats. CONCLUSIONS: These results demonstrate that in addition to eliciting peripheral insulin resistance, short-term CORT administration impairs insulin signaling in the rat hippocampus, effects that may contribute to the deleterious consequences of hypercortisolemic/hyperglycemic states observed in type 2 diabetes. PMID- 17426392 TI - Upper limb deep vein thrombosis: a literature review to streamline the protocol for management. AB - OBJECTIVE: The objective of this article is to provide up-to-date information about aetiology, pathogenesis, diagnostic modalities and treatment of upper limb deep vein thrombosis (ULDVT). METHODS: Generic terms including ULDVT, axillary subclavian DVT, and complications of central venous catheters were searched on electronic database. We analysed original studies, review articles and evaluation studies published over the last 25 years. RESULTS: Forty-seven studies on ULDVT encompassing 2,557 patients were evaluated. The incidence of ULDVT was quoted 1 4% of the total DVT. Primary ULDVT (20% of the total) was due to activity-related venous trauma. Secondary ULDVT (80% of the total) was due to central venous catheters and malignancy. Duplex ultrasound (sensitivity 78-100% and specificity 82-100%), contrast venography (gold standard) and magnetic resonance venography were the diagnostic tools used. Pulmonary embolism (2-35%) and post-thrombotic syndrome (7-46%) were the main sequelae. Anticoagulation was the universal intervention, giving 79% symptom relief (13.2% rethrombosis rate). Thrombolysis and/or percutaneous thrombectomy were used in 38% of cases for the management of ULDVT, giving 83% symptom relief (90% recanalization rate and 9% rethrombosis rate). Surgical decompression, venous angioplasty and superior vena cava filters were the main adjunctive interventions. CONCLUSION: ULDVT, although rare, is associated with considerable morbidity and mortality (29-40%) due to potential risks of pulmonary embolism, post-thrombotic syndrome and loss of vascular access. Simple anticoagulation is suitable for the majority of patients. Thrombolysis/thrombectomy is often successful but less frequently used. Surgical decompression, venous angioplasty and superior vena cava filters have some role in recurrent cases. An optimal management protocol can be established using a multimodality approach. PMID- 17426393 TI - Decreased treatment failure rates following duodenal release ferrous glycine sulfate in iron deficiency anemia associated with autoimmune gastritis and Helicobacter pylori gastritis. AB - BACKGROUND AND OBJECTIVES: Since gastric acidity and ascorbate play a critical role in the solubilization and reduction of iron for subsequent absorption, the achlorhydria associated with autoimmune and Helicobacter pylori gastritis may explain the poor response of such patients to oral iron treatment. In order to circumvent this problem, we explored the therapeutic potential of a duodenal formulation of ferrous glycine sulfate consisting of micropellets that do not dissolve at the acid environment of the stomach but, owing to their solubility at a higher pH, discharge their content directly into the duodenum. DESIGN AND METHODS: In a case-control study, the treatment results of 39 patients with iron deficiency anemia receiving a duodenal formulation of ferrous glycine sulfate (group A) were compared with the results of 39 patients receiving other oral iron compounds (group B). Autoimmune gastritis, H. pylori gastritis or both were present in over 75% of patients in each group. RESULTS: After 1 and 3 months of treatment, mean hemoglobin in group A increased from 9.5 +/- 1.2 to 11.2 +/- 1.3 and 12.8 +/- 1.3 g/dl, respectively. By comparison, in group B, the corresponding values increased from 9.3 +/- 1.3 to 10.2 +/- 1.5 (p = 0.019) and 11.1 +/- 1.7 g/dl (p = 0.022). A favorable response, defined as a more than 2 g/dl increase in basal hemoglobin or hemoglobin exceeding 12 g/dl, was obtained in 33 of 39 patients in group A compared with only 18 of 39 in group B (p = 0.009). Because of treatment failure, 14 patients in group B were subsequently referred for intravenous ferric sucrose therapy versus only 3 in group A (p < 0.0001). Conversely, of 5 patients in group A managed by intravenous iron prior to referral, 4 became independent of parenteral iron after starting the duodenal formulation of ferrous glycine sulfate. INTERPRETATION AND CONCLUSIONS: In patients with iron deficiency anemia associated with autoimmune and H. pylori gastritis with a high rate of refractoriness to oral iron treatment, satisfactory response to a duodenal formulation of ferrous glycine sulfate can be elicited in the vast majority of cases, obviating the need for expensive, inconvenient and occasionally risky intravenous iron administration. PMID- 17426394 TI - Utility technology in the assessment of the cut-off between a negative and a positive test in a caries prediction model. AB - The methodology for the assessment of a negative or positive test in caries prediction models has not received much attention. This study aims to explain how utility technology could be applied in a caries prediction model for the assessment of the cut-off between a negative and a positive test. In this study loss of utilities was assigned to false outcomes of the prediction test. A regression equation with past caries experience variables of 11-year-old children and caries increment in the 11- to 15-year forecast period as outcome was computed. The adjusted explained variance for caries increment was 0.45. Formulas were constructed for the loss of utilities for false-negative tests whereas false positive tests were corrected with a fixed loss of utilities. Each 11-year-old child of the 252 children was screened at various points of the regression equation. Loss of utilities was calculated for each child on the basis of the validation criterion, the outcome of the test and the actual caries increment of the child. The point on the regression equation with the least loss of utilities for the whole group was taken as the cut-off between a negative and a positive test. If the validation criterion for the prediction period was set on no caries, the prediction model resulted in a sensitivity of 84% and a specificity of 73%. This prediction model has potential when caries incidence is low and resources limited. PMID- 17426395 TI - Risk factors for a false positive test outcome in diagnosis of caries in approximal surfaces: impact of radiographic modality and observer characteristics. AB - The aim of this study was to evaluate the impact of image modality and observer characteristics on radiographic caries diagnosis with particular emphasis on the false positive (FP) test outcome. Eighty human teeth were radiographed by eight digital modalities and film. Eight observers scored caries lesions in each approximal tooth surface. Their use of image enhancement and time spent in assessing an image was recorded. Microscopy served as validation for the presence/absence of a lesion. Of 160 approximal surfaces, 63% were sound and 37% had lesions. Multivariate logistic regression analysis was performed with the FP test outcome as the dependent variable and the radiographic modality and observer's gender, experience, employment, image enhancement, and time use as the independent variables. The significant risk factors for an FP test outcome were: (1) gender: males scored fewer FP than females (OR 0.26, CI 0.18-0.36); (2) experience: observers with no experience with digital radiography had a six times higher risk for an FP (OR 6.09, CI 4.9-8.1); (3) radiographic modality: two of the digital systems had a significantly lower risk for an FP [a phosphor plate system (OR 0.55, CI 0.37-0.73) and a sensor-based system (OR 0.65, CI 0.46-0.93)] and one phosphor plate system had a higher risk (OR 1.57, CI 1.17-2.13) than film; (4) time use: more time was spent when scoring an FP (OR 1.05, CI 1.03 1.05) than for any other test outcome. Type of employment and the use of image enhancement had no significant impact on the risk for an FP diagnosis. PMID- 17426396 TI - Detection of approximal caries by clinical and radiographic examination in 9-year old Swedish children. AB - The aims were to determine caries prevalence in 9-year-olds belonging to a low caries prevalence population and to assess how accurately some commonly used risk factors/risk markers (predictors) can identify additional approximal caries as judged from bitewing examination (BW). One calibrated dentist examined 117 consecutive 9-year-olds from the inner city of Stockholm. The predictors were the number of clinically detected dfs and DFS, visible plaque, salivary mutans streptococci, tooth brushing habits, consumption of sugary products, parents' education, and (before BW) an overall clinical judgement by the examiner. Analyses of the data included sensitivity and specificity and a multiple logistic regression model. When BW was included, the mean DFS was 0.27 and the mean dfs 1.74. For approximal enamel and dentin lesions, the average gain from BW was 0.20 lesions for permanent first molars and 1.23 lesions for primary molars. Altogether, 48% of the children benefited from BW. The best predictors of additional approximal caries from BW were presence of caries (cavitation/dentin level) on occlusal surfaces of permanent first molars and the dentist's judgement based on clinical findings, both with a combined sensitivity and specificity of 134%. The only statistically significant variables in a logistic regression model were the dentist's judgement and the child's statement about regular tooth brushing. It is concluded that a sizeable proportion of 9-year-olds, representing a low-caries prevalence population, benefited from BW. However, the ability to identify these children from the predictors was limited. PMID- 17426397 TI - Reliability of quantitative laser fluorescence analysis of smooth surface lesions adjacent to the gingival tissues. AB - The aim of this study was to determine the impact of analysis patch border inclusion or exclusion on the reliability of quantitative laser fluorescence (QLF) analyses of lesions close to the gingival margin. Ninety-three lesions on the maxillary anterior teeth were imaged using a fluorescent capturing system. All the lesions were located on the gingival third of the teeth. One examiner undertook two analyses of the images 1 week apart and another examiner undertook a single analysis. Analyses were undertaken using QLF 2.00 g with four patch borders. Each border was assessed as active or inactive. QLF metrics DeltaF, DeltaQ and area were exported. Kappa statistics were used to measure the agreement of border inclusion between all three analyses, and intra-class correlation coefficients (ICCs) were used to determine the intra- and inter examiner reliability of the QLF metrics. Agreement on border exclusion was poor; with intra-examiner kappa of 0.48 and inter-examiner at 0.20. However, despite the inconsistencies in border exclusions the ICCs for each QLF metric were high; intra-examiner DeltaQ 0.91, DeltaF 0.80 and area 0.92; inter-examiner DeltaQ 0.86, DeltaF 0.68, area 0.88. Lesions adjacent to the gingival margin will often require a patch analysis border to be excluded in order to ensure a satisfactory reconstruction and thus accurate analysis. The decision to include or exclude a border and, if excluded, which border to select appears to be highly variable between examiners. Nevertheless, the QLF metrics appear to be robust as demonstrated by the high ICCs noted in this study. PMID- 17426398 TI - Effect of an alcohol-free, 1% chlorhexidine gel as an adjunct to a fluoridated dentifrice using an intraoral crown model. AB - BACKGROUND/AIMS: The use of chlorhexidine as a topically applied oral antiseptic is well documented; however, clinical studies examining the effects of chlorhexidine gel on in situ dental caries are limited. This study utilized an in situ caries model and a modified crossover design to examine whether the addition of a biweekly topical, alcohol-free, 1% chlorhexidine digluconate gel to a daily fluoridated dentifrice inhibited artificial caries in dental tissues better than the fluoridated dentifrice alone when compared to a nonfluoridated placebo dentifrice. METHODS: Thirty patients were recruited based on their need for a mandibular, full crown. Artificial caries lesions were created in extracted human teeth and enamel and root tissue sections 100 mum in thickness were characterized using polarized light microscopy. The sections were fixed in the crown and placed on the prepared tooth. The participants were assigned a placebo toothpaste, a toothpaste with 1,100 ppm F or a 1,100 ppm F toothpaste followed by 1 ml of 1% chlorhexidine gel at day 1 and day 14 (chlorhexidine+). Patients were instructed to brush twice daily for 4 weeks. Following each round, the sections in the crown were replaced with new sections. The sections were recharacterized and the mean changes were compared using ANOVA at alpha = 0.05. RESULTS: The chlorhexidine + Fdentifrice and the F dentifrice alone significantly reduced lesion area in enamel tissue when compared to the placebo dentifrice. Both treatments also inhibited lesion progression and initiation in root tissue better than control in this model system. Although the chlorhexidine+ group enhanced remineralization and inhibited lesion progression better than the F(-) dentifrice alone for all outcomes measured, the differences were not significant. CONCLUSIONS: The chlorhexidine, in conjunction with a fluoride dentifrice, was no more effective than the fluoride dentifrice alone. Further study is needed before this 1% alcohol-free chlorhexidine gel should be recommended as an adjunct to a fluoride dentifrice in the treatment of dental caries. PMID- 17426399 TI - Long-term effect of xylitol gum use on mutans streptococci in adults. AB - Many studies have shown the effects of chewing xylitol gum on mutans streptococci (MS) over short- and long-term periods in children; however, few studies have addressed long-term periods in adults. The objective of this investigation was to examine for 6 months the effects of chewing xylitol gum on MS in saliva and plaque in 127 adults (mean age 28.0 years). The participants were assigned to three groups according to gum type, in part taking preference for flavor into account and in part at random: xylitol (XYL), maltitol (MAL) and control (CR); 33, 34 and 27 subjects in each group, respectively, completed the trial. Daily gum use of the XYL and MAL groups was 7.9 and 7.1 g, respectively. MS levels, which declined significantly in saliva (p < 0.05) and plaque (p < 0.001) in the XYL group after 6 months, exhibited a significant increase in plaque in the MAL group (p < 0.001). Differences in relative changes of MS levels in plaque during the experimental period were significant between the XYL group and the CR (p < 0.05) and MAL groups (p < 0.001). Differences in relative change of amount of plaque during the experimental period were not statistically significant between the groups. The present study demonstrated that chewing xylitol gum for 6 months continued to inhibit the growth of mutans streptococci in adults. PMID- 17426400 TI - Ultrasonic determination of the effect of casein phosphopeptide-amorphous calcium phosphate paste on the demineralization of bovine dentin. AB - The purpose of this study was to investigate the demineralization of dentin by measuring changes in the velocity of the sonic longitudinal waves transmitted through this substrate. One group of samples was immersed in demineralization solution for 10 min twice a day and then stored in artificial saliva. Two additional groups of samples were treated with a solution of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste or a placebo paste without CPP-ACP before demineralization and a control group was stored in artificial saliva. The sonic velocity of the demineralized specimens was found to decrease significantly over time. No significant increase in sonic velocity was observed in specimens treated with CPP-ACP, suggesting that CPP-ACP acted to prevent demineralization. PMID- 17426401 TI - Effect of white cheese and sugarless yoghurt on dental plaque acidogenicity. AB - White cheese and sugarless yoghurt are frequently consumed traditional Turkish foods. The aim of this study was to assess their acidogenic potential when eaten alone or following a rinse with sucrose solution. Plaque pH was measured by a pH microelectrode at baseline to determine the resting plaque pH and at time intervals of 1-60 min after rinsing with 10% sucrose solution or eating the test food for 1 min. White cheese consumption for 1 min increased the plaque pH. When white cheese was eaten 5 min after rinsing with 10% sucrose solution, the plaque pH rose rapidly. After sugarless yoghurt consumption, the pH fell to a similar minimum as for sucrose after 5 min but returned to baseline levels after 30 instead of 60 min. Consumption of sugarless yoghurt after a sucrose rinse initially reduced the plaque pH further but did not affect the time taken for pH to regain baseline levels. PMID- 17426402 TI - Saliva-promoted adhesion of Streptococcus mutans MT8148 associates with dental plaque and caries experience. AB - Colonization of enamel surfaces by Streptococcus mutans is thought to be initiated by the attachment of bacteria to a saliva-derived conditioning film (acquired pellicle). However, the clinical relevance of the contribution of saliva-promoted S. mutans adhesion in biofilm formation has not yet been fully elucidated. The aim of this study was to correlate saliva-promoted S. mutans adhesion with biofilm formation in humans. We correlated all measurements of salivary factors and dental plaque formation in 70 healthy subjects. Dental plaque development after thorough professional teeth cleaning correlated positively with S. mutans adhesion onto saliva-coated hydroxyapatite pellets and the glycoprotein content of either parotid or whole saliva. Saliva-promoted S. mutans adhesion and glycoprotein content were also positively correlated with each other in parotid and whole saliva. By contrast, neither salivary mutans streptococci, Lactobacillus nor Candida correlated with biofilm formation. Parotid saliva-mediated S. mutans adhesion was significantly higher in 12 caries experienced (CE) subjects than in 9 caries-inexperienced (CI) subjects. Salivary S. mutans adhesion was significantly less (p < 0.01) in the CI group than in the CE group. In conclusion, the present findings suggest the initial S. mutans adhesion, modulated by salivary protein adsorption onto the enamel surface, as a possible correlate of susceptibility to dental plaque and caries. PMID- 17426403 TI - The influence of fluoride varnish on the attrition of dentine. AB - Previous investigations have shown the potential for fluoride to be protective in an abrasion/erosion laboratory model. The aim of this study was to investigate the effect of high concentrations of fluoride delivered in a varnish on attrition of dentine. Fifteen caries-free, intact lower third molar teeth were sectioned and the enamel removed by a water-cooled diamond disc. Polished dentine surfaces were divided into 8 areas, 4 of which were randomly covered with a high concentration fluoride varnish for 24 h. The samples were subjected to 5,000 cycles of attrition bathed under artificial saliva. Microhardness testing adjacent to the wear scars showed no statistical difference between the fluoride treated (71.42 KHN, SD 10.52) and control surfaces (72.66 KHN, SD 9.69). The volume of the wear scar was statistically greater for the fluoride-treated surface at 9.6 microm(3) (SD 4.92) and 8.13 microm(3) (SD 5.54) for the control areas (p = 0.029). The low pH of the fluoride varnish appears to have increased the amount of wear from attrition in this laboratory study. PMID- 17426404 TI - Surface layer erosion of natural caries lesions with phosphoric and hydrochloric acid gels in preparation for resin infiltration. AB - The infiltration of proximal enamel lesions with low-viscosity light curing resins could be a viable approach to stop lesion progression. However, penetration of sealant might be hampered by the comparatively highly mineralized surface layers of natural lesions. Therefore, the aim of this study was to compare the efficacy of three different etching gels in removing the surface layer in various etching times. Extracted human molars and premolars showing proximal white spot lesions were cut across the demineralized areas. Ninety-six lesions expected from visual examination to be confined to the outer enamel (C1) were selected. The cut surface and half of each lesion were varnished, thus serving as control. Subsequently, the lesions were etched with either phosphoric (37%) or hydrochloric (5 or 15%) acid gel for 30-120 s (n = 8/group). Specimens were examined using confocal microscopy and transversal microradiography. Surface layer reduction was significantly increased in lesions etched with 15% HCl gel for 90 and 120 s compared to those etched with H(3)PO(4) gel for 30-120 s (p < 0.05). No significant differences regarding the depths of erosion in the lesions compared to sound enamel could be observed (p > 0.05). An effective reduction in the surface layer of natural enamel caries can be achieved by etching with 15% hydrochloric acid gel for 90-120 s. PMID- 17426405 TI - Detection of proximal caries in vitro using standard and task-specific enhanced images from a storage phosphor plate system. AB - Eight dentists evaluated 72 proximal surfaces of premolars with respect to all caries lesions and to lesions into dentine in digital images from a storage phosphor plate system. The images were processed in four different ways: (1) the default algorithm of the standard imaging system (DF); (2) an algorithm correcting for attenuation and visual response (AV), and two proprietary caries specific enhancement algorithms (3) K1, and (4) K2, respectively. The lesions were validated by histological examination. There were no significant differences in the areas under the receiver operating characteristic curves between differently processed radiographs for the categories of all caries lesions and caries into dentine. PMID- 17426406 TI - The effect of saliva on the formation of KOH-soluble fluoride after topical application of amine fluoride solutions of varying fluoride concentration and pH. AB - The effect of saliva on the amount of KOH-soluble fluoride formed on a sound enamel surface after application of amine fluoride solution of varying fluoride concentrations (1, 0.5 and 0.25% F) and pH (5.3, 4.5 and 4.0) was examined in an in vitro study. The saliva pretreatment increased the amount of KOH-soluble fluoride at the highest pH value. For any given fluoride concentration, the presence of saliva did not influence the amount of KOH-soluble fluoride. These data suggest that saliva could enhance the amount of KOH-soluble fluoride if the topical fluoride preparations are not highly acid. PMID- 17426407 TI - Lack of effect of prior treatment with fluoride on genotoxicity of two chemical agents in vitro. AB - The goal of this study was to investigate the ability of fluoride to modulate the genotoxic effects induced by the oxidative agent hydrogen peroxide (H2O2) and the alkylating agent methyl methanesulfonate (MMS) in vitro by the single-cell gel (comet) assay. Chinese hamster ovary cells were exposed in culture for 1 h at 37 degrees C to sodium fluoride at 7-100 microg/ml. NaF-treated and control cells were then incubated with 0-10 microM MMS in phosphate-buffered saline (PBS) for 15 min at 37 degrees C, or 0-100 microM H2O2 in distilled water for 5 min on ice. Negative control cells were treated with PBS for 1 h at 37 degrees C. Clear concentration-related effects were observed for the two genotoxins. Increase of DNA damage induced by either MMS or H2O2 was not significantly altered by pretreatment with NaF. The data indicate that NaF does not modulate alkylation induced genotoxicity or oxidative DNA damage as measured by the single-cell gel (comet) assay. PMID- 17426408 TI - Current approaches to the diagnosis and treatment of polycystic ovarian syndrome in youth. AB - Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in reproductive-age women. It often presents during late adolescence but in some cases certain features are evident even before menarche. PCOS is a spectrum of disorders with any combination of oligo/anovulation, clinical and/or biochemical evidence of androgen excess, obesity, insulin resistance and polycystic ovaries on ultrasound. The pathogenesis is unknown; however, it is a complex multigenetic disorder where disordered gonadotropin release, dysregulation of steroidogenesis, hyperinsulinism and insulin resistance play a role. The diagnosis is based on a typical physical exam (acne, hirsutism, obesity, and acanthosis nigricans) and laboratory evidence of hyperandrogenism, such as elevated free testosterone, androstenedione and dehydroepiandrosterone sulfate (DHEAS), decreased sex hormone binding globulin (SHBG) and increased luteinizing hormone (LH). An ovarian ultrasound may detect the multiple cysts. Secondary causes of PCOS need to be excluded. There are several classes of medications correcting different parameters of PCOS that can be used alone or in combination. Oral contraceptive therapy is used to reduce androgen and LH levels with resultant improvement in acne and hirsutism, and the induction of regular menses. Antiandrogens are usually required for a substantial improvement in hirsutism score. Insulin sensitizers such as metformin are a new class of drugs utilized in treatment of PCOS. By improving insulin sensitivity and decreasing insulin levels, they improve the unfavorable metabolic profile of patients with PCOS. Metformin also helps to increase SHBG, decrease androgen levels and induce ovulation. Despite all the available medications, life-style changes are the mainstay of therapy as weight loss and exercise improve all parameters of PCOS without the potential side effects of medication. PMID- 17426409 TI - Electromyography in evaluation of pain after different types of tonsillectomy: prospective randomized study. AB - BACKGROUND: Combined surface electromyography (sEMG) and visual analogue scale (VAS) pain assessment was performed on randomly assigned adults following tonsillectomy to research the effect of fibrin sealant (Group 1, n = 25), CO(2) laser (Group 2, n = 18) and electrocoagulation (Group 3, n = 40) on postsurgical pain. METHODS: Timing and amplitude of sEMG activity of masseter, infrahyoid and submandibular muscles were measured during swallowing simultaneously with VAS assessment and compared to normative database. RESULTS: Postsurgical electric activity of masseter and infrahyoid was significantly higher in comparison with normative database. sEMG and VAS data showed that tonsillectomy ended with fibrin sealant caused less pain than laser or electrocoagulation techniques with no correlation between the VAS score and duration of swallows. CONCLUSION: Tonsillectomy affects muscle activity significantly by involving additional muscles (infrahyoid) in swallowing. Combined sEMG and VAS data indicated electrocautery as the most painful, laser as less painful, and fibrin sealant as the least painful technique. PMID- 17426410 TI - Effects of mitemcinal (GM-611), an acid-resistant nonpeptide motilin receptor agonist, on the gastrointestinal contractile activity in conscious dogs. AB - The effects of mitemcinal (GM-611) on the gastrointestinal contractile activity were investigated using chronically implanted force transducers in conscious dogs and were compared with the effects of porcine motilin (pMTL), EM-523 and EM-574. In the interdigestive state, intravenous and oral administration of mitemcinal, EM-523 and EM-574 induced the gastrointestinal contractile activity in a manner similar to pMTL. The contractile activity caused by mitemcinal was suppressed by continuous intravenous infusion of a motilin receptor antagonist. In the digestive state, intravenous and oral administration of mitemcinal, EM-523 and EM 574 also stimulated the gastrointestinal contractile activity. Mitemcinal, EM-523 and EM-574 given intravenously increased the gastric contractile activity in a similar dose range; however, mitemcinal was approximately 10 times more potent than EM-523 and EM-574 when administered orally in the digestive state. These results indicate that the mitemcinal-induced gastrointestinal contractile activity operates via motilin receptors and possesses a higher activity than EM 523 and EM-574 when administered orally in conscious dogs in the digestive state. Mitemcinal may therefore be useful in the treatment of several gastrointestinal disorders involving dysmotility, such as gastroparesis and functional dyspepsia, even when administered orally. PMID- 17426411 TI - Demoralization and Psychotherapy: A Tribute to Jerome D. Frank, MD, PhD (1909 2005). PMID- 17426412 TI - Sexual symptoms in endocrine diseases: psychosomatic perspectives. AB - BACKGROUND: Not only the most frequent causes of endocrine sexual dysfunction, such as hypogonadism and hyperprolactinemia, but almost all extragonadal endocrinopathies (hyper- and hypothyroidism, hyper- and hypocortisolism, steroidal secreting tumors, etc.) may have a greater or lesser effect on sexual function. METHODS: We analyzed scientific literature on the correlations between hormones and sexual behavior, analyzing the most important issue from a practical point of view. The aim of this review article was thus to summarize the sexual symptoms that may be observed with endocrine diseases. RESULTS: Hormones directly or indirectly regulate all human sexual functions (desire, erection/lubrication, ejaculation, orgasm). Some sexual symptoms may occur as a psychosomatic consequence of hormonal impairment. However, in other cases, endocrine failure may be generated by the psychosomatic involvement. CONCLUSIONS: The endocrinologist, as an expert in body chemistry, is ideally positioned to identify and evaluate the full range of medical, physical, and psychiatric problems disrupting sexual function. PMID- 17426413 TI - How does our brain constitute defense mechanisms? First-person neuroscience and psychoanalysis. AB - Current progress in the cognitive and affective neurosciences is constantly influencing the development of psychoanalytic theory and practice. However, despite the emerging dialogue between neuroscience and psychoanalysis, the neuronal processes underlying psychoanalytic constructs such as defense mechanisms remain unclear. One of the main problems in investigating the psychodynamic-neuronal relationship consists in systematically linking the individual contents of first-person subjective experience to third-person observation of neuronal states. We therefore introduced an appropriate methodological strategy, 'first-person neuroscience', which aims at developing methods for systematically linking first- and third-person data. The utility of first-person neuroscience can be demonstrated by the example of the defense mechanism of sensorimotor regression as paradigmatically observed in catatonia. Combined psychodynamic and imaging studies suggest that sensorimotor regression might be associated with dysfunction in the neural network including the orbitofrontal, the medial prefrontal and the premotor cortices. In general sensorimotor regression and other defense mechanisms are psychoanalytic constructs that are hypothesized to be complex emotional-cognitive constellations. In this paper we suggest that specific functional mechanisms which integrate neuronal activity across several brain regions (i.e. neuronal integration) are the physiological substrates of defense mechanisms. We conclude that first-person neuroscience could be an appropriate methodological strategy for opening the door to a better understanding of the neuronal processes of defense mechanisms and their modulation in psychoanalytic psychotherapy. PMID- 17426414 TI - Which panic disorder patients benefit from which treatment: cognitive therapy or antidepressants? AB - BACKGROUND: Beliefs about the controllability of a disorder may be relevant in the causation, maintenance and treatment of disorders. We investigated whether congruence between patients' beliefs about controllability of a panic disorder and the type of treatment provided predicted outcome. METHODS: The differential effectiveness of cognitive therapy and antidepressant treatment (paroxetine or clomipramine) was investigated in a sample of 129 panic disorder patients in a 12 week, pretest posttest placebo-controlled study. Panic frequency, agoraphobic avoidance, anxiety, depression, and disability were measured with various validated interviewer and self-report measures. Beliefs about controllability were measured with the Multidimensional Anxiety Locus of Control Scale measuring an internal, chance, therapist and medication locus of control. In order to analyze aptitude-treatment interactions a new strategy called the Regression Trunk Approach was used in addition to classical hierarchical multiple regression analysis. RESULTS: Using the Regression Trunk Approach we found that locus of control orientation (LOC) predicted the differential effectiveness of cognitive therapy. Those patients with a medium internal LOC who received cognitive therapy performed significantly better than all patients who received a placebo pill on 8 of the 10 outcome variables. We did not find a differential LOC effect for antidepressant treatment. No evidence for aptitude-treatment interactions using hierarchical multiple regression analysis was found. CONCLUSIONS: Moderately strong beliefs about self-control of panic disorder congruent with the cognitive intervention provided seem to moderate treatment effectiveness. Future studies must be more attentive to the nonlinear effects of patient characteristics on the outcome of different types of treatments. PMID- 17426415 TI - Intrapersonal conflict in goals and values of patients with unipolar depression. AB - BACKGROUND: Although increased conflicts between attitudes and beliefs about certain goals or values are often discussed as important factors in depression, there are only few empirical studies investigating these relations among patients with depressive disorders. METHODS: In the present study, we used the Intrapersonal Conflict Test to assess cognitive inconsistencies in goals or values. A total of 53 inpatients with unipolar depression and 24 nondepressed controls (inpatients of an internal and a surgery ward) participated in the study. In addition to the Intrapersonal Conflict Test, patients completed the Beck Depression Inventory, the Dysfunctional Attitude Scale, the Inventory of Interpersonal Problems as well as the Problem Solving Inventory. RESULTS: Compared with controls, patients with depressive disorders showed significantly higher scores for global inconsistencies, inconsistencies within different goals/values, as well as between goals/values and their perceived realization. Significant correlations were found between conflict measures and the Inventory of Interpersonal Problems, as well as the Problem Solving Inventory. Path analyses show that group differences in intrapersonal conflicts were partially mediated by interpersonal problems but not by depressive symptoms or cognitive vulnerability factors. CONCLUSIONS: Given the cross-sectional design of the study, the findings of this exploratory study do not allow for conclusions regarding the role of intrapersonal conflicts in the development and course of depression. Nevertheless, the high levels of intrapersonal conflicts observed in the study suggest that inconsistencies in goals or values should be considered in the psychological treatment of depression. PMID- 17426416 TI - Is a full recovery possible after cognitive behavioural therapy for chronic fatigue syndrome? AB - BACKGROUND: Cognitive behavioural therapy (CBT) for chronic fatigue syndrome (CFS) leads to a decrease in symptoms and disabilities. There is controversy about the nature of the change following treatment; some suggest that patients improve by learning to adapt to a chronic condition, others think that recovery is possible. The objective of this study was to find out whether recovery from CFS is possible after CBT. METHODS: The outcome of a cohort of 96 patients treated for CFS with CBT was studied. The definition of recovery was based on the absence of the criteria for CFS set up by the Center for Disease Control (CDC), but also took into account the perception of the patients' fatigue and their own health. Data from healthy population norms were used in calculating conservative thresholds for recovery. RESULTS: After treatment, 69% of the patients no longer met the CDC criteria for CFS. The percentage of recovered patients depended on the criteria used for recovery. Using the most comprehensive definition of recovery, 23% of the patients fully recovered. Fewer patients with a co-morbid medical condition recovered. CONCLUSION: Significant improvement following CBT is probable and a full recovery is possible. Sharing this information with patients can raise the expectations of the treatment, which may enhance outcomes without raising false hopes. PMID- 17426417 TI - Low control at work and the risk of suicide in Japanese men: a prospective cohort study. AB - BACKGROUND: Although adverse psychosocial job characteristics are suspected predictors of suicide death, prospective studies based on established stress instruments are limited. METHODS: In a multicenter community-based Japanese cohort study, we prospectively investigated the association between psychosocial job characteristics and the risk of death from suicide among male workers. Baseline examination was conducted from 1992 to 1995 to determine the socioeconomic, behavioural and biological variables in addition to the psychosocial job characteristics of 3,125 male workers aged 65 and under and free from major illness. Low job control and high job demands were measured as adverse psychosocial job characteristics according to a job demand-control model questionnaire. Suicide deaths were identified using the Cause-of-Death Register. RESULTS: During the 9-year follow-up, 14 suicides were identified. The suicide death rate was 48.1 per 100,000 person years. Multivariate analysis revealed a more than fourfold increase in the risk of suicide among men with low control at work (relative risk: 4.10; 95% confidence interval: 1.31-12.83) compared with counterpart men after adjustment for age, marital status, educational attainment, occupation, smoking status, alcohol consumption, total cholesterol level, and study area. Job demands were not associated with risk of death from suicide. CONCLUSIONS: By using a job demand-control model questionnaire, low control at work was revealed as a predictor of suicide death among Japanese male workers. The finding implies that job redesign aimed at increased worker control could be a worthwhile strategy in preventing, or at least reducing, the risk of suicide death. PMID- 17426418 TI - Assessing alexithymia and type A behavior in coronary heart disease patients: a multimethod approach. AB - BACKGROUND: Despite increasing emphasis on using multiple methods to assess personality constructs in psychosomatic research, previous investigations of relations between alexithymia and type A behavior (TAB) have been limited by the use of single methods of measurement and almost no attempt to assess subcomponents of TAB. The aims of this study were to (1) evaluate levels of agreement between structured interview assessments of alexithymia, TAB, hostility, and time urgency and well-established self-report measures of these constructs, and (2) explore relations between alexithymia and TAB and its subcomponents in patients with coronary heart disease (CHD). METHODS: 62 CHD patients were investigated 6 weeks after coronary angioplasty. Alexithymia was assessed with the Diagnostic Criteria for Psychosomatic Research (DCPR) and the 20-item Toronto Alexithymia Scale (TAS-20). TAB was assessed with the DCPR and the Short Form of the Jenkins Activity Survey Type A scale (JAS-SF). Time urgency was assessed with the DCPR and the Speed/Impatience scale of the Jenkins Activity Survey (JAS-S), and hostility was assessed with the DCPR and the Hostility subscale of the Revised Symptom Checklist-90 (SCL-HOS). RESULTS: The DCPR classifications showed reasonably high levels of agreement with the TAS-20 and JAS-SF classifications of alexithymia and TAB, but lower levels of agreement in identifying patients with high hostility on the SCL-HOS and high time urgency on the JAS-S. Alexithymia measured by both the DCPR and the TAS-20 was unrelated to both self-report and structured interview measures of TAB, hostility, and time urgency. CONCLUSIONS: The DCPR is a suitable screening instrument for assessing alexithymia and TAB, although the two constructs are unrelated. PMID- 17426419 TI - [Strategy considerations in genome cohort construction in Korea]. AB - Focusing on complex diseases of public health significance, strategic issues regarding the on-going Korean Genome Cohort were reviewed: target size and diseases, measurements, study design issues, and followup strategy of the cohort. Considering the epidemiologic characteristics of Korean population as well as strengths and drawbacks of current research environment, we tried to tailor the experience of other existing cohorts into proposals for this Korean study. Currently 100,000 individuals have been participating the new Genome Cohort in Korea. Target size of de novo collection is recommended to be set as between 300,000 to 500,000. This target size would allow acceptable power to detect genetic and environmental factors of moderate effect size and possible interactions between them. Family units and/or special subgroups are recommended to parallel main body of adult individuals to increase the overall efficiency of the study. Given that response rate to the conventional re-contact method may not be satisfactory, successful follow-up is the main key to the achievement of the Korean Genome Cohort. Access to the central database such as National Health Insurance data can provide enormous potential for near-complete case detection. Efforts to build consensus amongst scientists from broad fields and stakeholders are crucial to unleash the centralized database as well as to refine the commitment of this national project. PMID- 17426420 TI - [High throughput genotyping for genomic cohort study]. AB - Human Genome Project (HGP) could unveil the secrets of human being by a long script of genetic codes, which enabled us to get access to mine the cause of diseases more efficiently. Two wheels for HGP, bioinformatics and high throughput technology are essential techniques for the genomic medicine. While microarray platforms are still evolving, we can screen more than 500,000 genotypes at once. Even we can sequence the whole genome of an organism within a day. Because the future medicine will focus on the genetic susceptibility of individuals, we need to find genetic variations of each person by efficient genotyping methods. PMID- 17426421 TI - [Statistical issues in genomic cohort studies]. AB - When conducting large-scale cohort studies, numerous statistical issues arise from the range of study design, data collection, data analysis and interpretation. In genomic cohort studies, these statistical problems become more complicated, which need to be carefully dealt with. Rapid technical advances in genomic studies produce enormous amount of data to be analyzed and traditional statistical methods are no longer sufficient to handle these data. In this paper, we reviewed several important statistical issues that occur frequently in large scale genomic cohort studies, including measurement error and its relevant correction methods, cost-efficient design strategy for main cohort and validation studies, inflated Type I error, gene-gene and gene-environment interaction and time-varying hazard ratios. It is very important to employ appropriate statistical methods in order to make the best use of valuable cohort data and produce valid and reliable study results. PMID- 17426422 TI - [A review of power and sample size estimation in genomewide association studies]. AB - Power and sample size estimation is one of the crucially important steps in planning a genetic association study to achieve the ultimate goal, identifying candidate genes for disease susceptibility, by designing the study in such a way as to maximize the success possibility and minimize the cost. Here we review the optimal two-stage genotyping designs for genomewide association studies recently investigated by Wang et al(2006). We review two mathematical frameworks most commonly used to compute power in genetic association studies prior to the main study: Monte-Carlo and non-central chi-square estimates. Statistical powers are computed by these two approaches for case-control genotypic tests under one-stage direct association study design. Then we discuss how the linkage disequilibrium strength affects power and sample size, and how to use empirically-derived distributions of important parameters for power calculations. We provide useful information on publicly available software developed to compute power and sample size for various study designs. PMID- 17426423 TI - [Ethical considerations in genomic cohort study]. AB - During the last decade, genomic cohort study has been developed in many countries by linking health data and genetic data in stored samples. Genomic cohort study is expected to find key genetic components that contribute to common diseases, thereby promising great advance in genome medicine. While many countries endeavor to build biobank systems, biobank-based genome research has raised important ethical concerns including genetic privacy, confidentiality, discrimination, and informed consent. Informed consent for biobank poses an important question: whether true informed consent is possible in population-based genomic cohort research where the nature of future studies is unforeseeable when consent is obtained. Due to the sensitive character of genetic information, protecting privacy and keeping confidentiality become important topics. To minimize ethical problems and achieve scientific goals to its maximum degree, each country strives to build population-based genomic cohort research project, by organizing public consultation, trying public and expert consensus in research, and providing safeguards to protect privacy and confidentiality. PMID- 17426424 TI - [Impact of an early hospital arrival on treatment outcomes in acute ischemic stroke patients]. AB - OBJECTIVES: Recent educational efforts have concentrated on patient's early hospital arrival after symptom onset. The purpose of this study was to evaluate the time interval between symptom onset and hospital arrival and to investigate its relation with clinical outcomes for patients with acute ischemic stroke. METHODS: A prospective registry of patients with signs or symptoms of acute ischemic stroke, admitted to the OO Medical Center through emergency room, was established from September 2003 to December 2004. The interval between symptom onset and hospital arrival was recorded for each eligible patient and analyzed together with clinical characteristics, medication type, severity of neurologic deficits, and functional outcomes. RESULTS: Based on the data of 256 patients, the median interval between symptom onset and hospital arrival was 13 hours, and 22% of patients were admitted to the hospital within 3 hours after symptom onset. Patients of not-mild initial severity and functional status showed significant differences between arrival hours of 0-3 and later than 3 in terms of their functional outcomes on discharge. Logistic regression models also showed that arrival within 3 hours was a significant factor influencing functional outcome (OR=5.6; 95% CI=2.1, 15.0), in addition to patient's initial severity, old age, cardioembolism subtype, and referral to another hospital. CONCLUSIONS: The time interval between symptom onset and hospital arrival significantly influenced treatment outcome for patients with acute ischemic stroke, even after controlling for other significant clinical characteristics. The findings provided initiatives for early hospital arrival of patients and improvement of emergency medical system. PMID- 17426425 TI - [Association of social support and social activity with physical functioning in older persons]. AB - OBJECTIVES: According to Rowe and Kahn (1998), successful aging is the combination of a low probability of disease, high functioning, and active engagement with life. The purpose of this study was to assess the relationship between active engagement with life and functioning among the community-dwelling elderly. METHODS: Data were collected from Wave 2 of the Suwon Longitudinal Aging Study (SLAS), consisting of a sample of 645 persons aged 65 and older living in the community. A social activity checklist and social support inventory were used as measures of engagement with life, along with the Physical Functioning (PF) scale as a measure of functioning. The effects of social support and social activity on physical functioning, taking into account the covariates, were analyzed by hierarchical linear regression analysis. RESULTS: Maintenance of social activity and social support were significantly associated with higher physical function, after adjusting for sociodemographic and health-related covariates. Social support appeared to be more prominent than social activity in predicting physical functioning. CONCLUSIONS: Social support and social activity are potentially modifiable factors associated with physical function in older persons. Studies examining the role social engagement may play in preventing disability are warranted. PMID- 17426426 TI - Using tobit regression analysis to further understand the association of youth alcohol problems with depression and parental factors among Korean adolescent females. AB - OBJECTIVES: This study characterized the extent to which youth depressive symptoms, parental alcohol problems, and parental drinking account for differences in alcohol-related problems among a large sample of adolescent females. METHODS: The stratified sample consists of 2077 adolescent females from twelve female-only high schools located in a large metropolitan city in the Republic of Korea. Students completed a questionnaire about alcohol use and alcohol problems, their parents' alcohol problems, and a number of risk and protective factors. Data were analyzed using tobit regression analyses to better characterize the associations among variables. RESULTS: Almost two-thirds of students who consume alcohol had experienced at least one to two alcohol-related problems in their lives and 54.6% reported at least one current symptom of depression, with nearly one-third reporting two depressive symptoms. Two-thirds of the students indicated that at least one parent had an alcohol-related problem, and that approximately 29% had experienced several problems. Results of tobit regression analyses indicate that youth alcohol-related problems are positively associated with depressive symptoms (p<0.01) and parent drinking problems (p<0.05). Parental drinking is no longer significant when the variable parental attention is added to the model. Decomposition of the tobit parameters shows that for every unit of increase in depressive symptoms and in parent drinking problems, the probability of a youth experiencing alcohol problems increases by 6% and 1%, respectively. For every unit of increase in parental attention, the probability of youth experiencing drinking problems decreases by 5%. CONCLUSIONS: This study presents evidence that alcohol-related problems and depressive symptoms are highly prevalent among adolescent females. Although a comprehensive public health approach is needed to address drinking and mental health problems, different interventions are needed to target factors associated with initiation of alcohol problems and those associated with increased alcohol problems among those who already began experiencing such problems. PMID- 17426427 TI - [The determinants of purchasing private health insurance in Korean cancer patients]. AB - OBJECTIVES: The aim of this study is to identify factors determining the purchase of private health insurance under the mandatory National Health Insurance(NHI) system in Korea. METHODS: The data were collected by the National Cancer Center in Korea. It includes cancer patients who were newly diagnosed with stomach (ICD code, C16), lung(C33-C34), liver (C22), colorectal cancer(C18-C20) or breast(C50) cancer. Data were gathered from the hospital Order Communication System (OCS), medical records, and face-to-face interviews, using a structured questionnaire. Clinical, socio-demographic and private health insurance related factors were also gathered. RESULTS: Overall, 43.9% of patients had purchased one or more private health insurance schemes related to cancer, with an average monthly premium of won 65,311 and an average benefit amount of won 19 million. Females, younger aged, high income earners, national health insurers and metropolitan citizens were more likely to purchase private health insurance than their counterparts. CONCLUSIONS: About half of Korean people have supplementary private health insurance and their benefits are sufficient to cover the out-of-pocket fees required for cancer treatment, but inequality remains in the purchase of private health insurance. Further studies are needed to investigate the impacts of private health insurance on NHI, and the relationship between cancer patients' burden and benefits. PMID- 17426428 TI - Developmental toxicity by exposure to bisphenol A diglycidyl ether during gestation and lactation period in Sprague-Dawley male rats. AB - OBJECTIVES: Bisphenol A diglycidyl ether (BADGE) is the major component in commercial liquid epoxy resins, which are manufactured by co-reacting bisphenol A with epichlorohydrin. This study was performed to show the developmental effects of prenatal and postnatal exposures to BADGE in male rat offspring. METHODS: Mated female rats were divided into four groups, each containing 12 rats. The dosing solutions were prepared by thoroughly mixing BADGE in corn oil at the 0, 375, 1500 and 3000 mg/kg/day concentrations. Mated females were dosed once daily by oral gavage on gestation day (GD) 6 - 20 and postnatal day (PND) 0 - 21. Pregnant female dams were observed general symptoms and body weight. Also, male pups were observed the general symptoms, body weight, developmental parameters (e.g. anogenital distance, pina detachment, incisor eruption, nipple retention, eye opening, testis descent), organ pathologic changes and hormone levels of plasma. RESULTS: Pregnant rats treated with BADGE died at a rate of about 70% in the 1500 mg/kg/day group and all rats treated with 3000 mg/kg/day died. Body weight, for male pups treated with doses of 375 mg/kg/day, was significantly lower than in the control group at PND 42, 56, and 63 (p<0.05). Evaluation of body characteristics including; separation of auricle, eruption of incisor, separation of eyelid, nipple retention, descent of testis, and separation of the prepuce in the BADGE treated group showed no difference in comparisons with the control group. AGD and adjusted AGD (mm/kg) for general developmental items in BADGE 375 mg/kg/day treated pups tended to be longer than in controls, however, these differences were not statistically significant. Relative weights of adrenal gland, lung (p<0.05), brain, epididymis, prostate, and testis (p<0.01) were heavier than in control in measures at PND 9 weeks. There were no significant changes in comparisons of histological findings of these organs. Loss of spermatids was observed in the seminiferous tubule at PND 9 weeks, but no weight changes were observed. The plasma estrogen levels were similar in the control and treatment groups at PND 3, 6 and 9 weeks. The plasma testosterone levels in the control group tended to increase with age. However, in the BADGE 375 mg/kg/day treated male pups it did not tend to increase. CONCLUSIONS: These findings suggest that BADGE is a chemical that has developmental effects consistent with it being an endocrine disruptor. PMID- 17426429 TI - [Change of cognitive function and associated factors among the rural elderly: a 5 year follow-up study]. AB - OBJECTIVES: This prospective population-based cohort study was conducted to evaluate the risk factors of cognitive impairment and the degree of cognitive function change through a 5-year follow-up. METHODS: The baseline and follow-up surveys were conducted in 1998 and 2003, respectively. Among 176 subjects who had normal cognitive function in the baseline study, 136 were followed up for 5 years. The cognitive function was investigated using the Korean version of the Mini-Mental State Examination (MMSE-K). The collected data were analyzed using SPSS and Stata. RESULTS: Of the 136 subjects analyzed, 25 (18.4%) were cognitively impaired. Old age and low social support in the baseline survey were risk factors for cognitive impairment after 5 years. In the generalized estimating equation for 128 subjects except severe cognitive impairment about the contributing factors of cognitive function change, the interval of 5 years decreased MMSE-K score by 1.02 and the cognitive function was adversely affected with increasing age, decreasing education and decreasing social support. CONCLUSIONS: Although the study population was small, it was considered that the study results can be used to develop a community-based prevention system for cognitive impairment. PMID- 17426430 TI - [General population time trade-off values for 42 EQ-5D health states in South Korea]. AB - OBJECTIVES: This study was conducted to elicit quality weights for 42 EQ-5D health states with the time trade-off (TTO) method from the general population of South Korea. METHODS: We selected the same EQ-5D health states as those in the UK MVH study. The Korean version of EQ-5D questionnaire and TTO method were used for the valuation process. We interviewed 500 people as a representative sample of the general population in Seoul and Gyeonggi-do. The result was compared with those from UK, Japan, and USA by Spearman's rank correlation and t-test. RESULTS: TTO values for 42 EQ-5D health states and 'unconscious' state were obtained from the general South Korean population. The best one was '11112' state and the worst one was 'unconscious' state. The states worse than death were '33323', '33333', and 'unconscious' states, which had negative TTO values. There was a strong correlation between TTO values of the EQ-5D health states and those of their corresponding states from UK, Japan, and USA (Spearman's correlation coefficient: 0.885, 0.882, and 0.944, respectively, p <0.001). However, absolute TTO values of most EQ-5D health states were significantly different from those of their corresponding states in other foreign studies (UK: 41/42, USA: 32/42, Japan: 15/17). CONCLUSIONS: We found that the Korean general population TTO values for EQ-5D health states were different from those of other foreign studies, suggesting that a specific Korean valuation set should be developed and used for economic evaluation studies in South Korea. PMID- 17426431 TI - [The levels of psychosocial stress, job stress and related factors of medical doctors practicing at local clinics]. AB - OBJECTIVES: This study was conducted to investigate the levels of psychosocial stress, job stress and their related factors among medical doctors practicing at local clinics. METHODS: A survey using a self administered questionnaire was administered to 1,456 doctors practicing at private clinics via post for 2 months (2006. 1 - 2006. 3). Psychosocial stress, job stress,demographic factors, job related factors and health related behaviors were investigated. Among the eligible study population, the respondents were 428 doctors (29.4%). RESULTS: The average scores of psychosocial stress and job stress were 2.19 and 3.13, respectively. The levels of psychosocial stress and job stress were statistically lower in older respondents, those who worked shorter or who were more satisfied with their job, and those with higher socioeconomic status. The level of psychosocial stress was related with smoking status, drinking status and exercise. The level of job stress was related with smoking status and exercise. In multiple linear regression analysis using psychosocial stress as a dependent variable, age, working hours per day, job satisfaction and perception on socioeconomic status were significant independent variables. In analysis using job stress as a dependent variable, age, working hours per day and job satisfaction were significant independent variables. CONCLUSIONS: Stress affects the doctor-patient relationship, productivity and overall health level of people. Therefore, it is important to manage and relieve the stress of doctors. It is suggested that more advanced studies on stress level and related factors and ways to improve the stress and health related behaviors of medical doctors should be conducted. PMID- 17426432 TI - [The current trend of avian influenza viruses in bioinformatics research]. AB - OBJECTIVES: Since the first human infection from avian influenza was reported in Hong Kong in 1997, many Asian countries have confirmed outbreaks of highly pathogenic H5N1 avian influenza viruses. In addition to Asian countries, the EU authorities also held an urgent meeting in February 2006 at which it was agreed that Europe could also become the next target for H5N1 avian influenza in the near future. In this paper, we provide the general and applicable information on the avian influenza in the bioinformatics field to assist future studies in preventive medicine. METHODS: We introduced some up-to-date analytical tools in bioinformatics research, and discussed the current trends of avian influenza outbreaks. Among the bioinformatics methods, we focused our interests on two topics: pattern analysis using the secondary database of avian influenza, and structural analysis using the molecular dynamics simulations in vaccine design. RESULTS: Use of the public genome databases available in the bioinformatics field enabled intensive analysis of the genetic patterns. Moreover, molecular dynamic simulations have also undergone remarkable development on the basis of the high performance supercomputing infrastructure these days. CONCLUSIONS: The bioinformatics techniques we introduced in this study may be useful in preventive medicine, especially in vaccine and drug discovery. PMID- 17426433 TI - The impacts of obesity on psychological well-being: a cross-sectional study about depressive mood and quality of life. AB - OBJECTIVES: The aim of this study was to assess whether individuals who visit clinics to ask medical help for obesity treatment depict comparable levels of depression, body dissatisfaction, eating psychopathology and lower quality of life. METHODS: This is a cross sectional study with 534 females who sought treatment for their obesity or overweight being recruited in seven clinical units in Seoul, Korea. The patients group was divided into two groups. The group 1 consisted of the patients with BMI >25 kg/m(2). The women who showed BMI < or =25 kg/m(2) among patients recruited for this study were classified as the group 2. The control group (group 3) was composed of 398 healthy females who have never tried to lose weight. RESULTS: We found that group 1 had higher frequency of more than moderate level of depression than group 2 and group 3 did. Both patients groups showed greater eating disordered attitudes and behaviors regardless of obese condition than the control group. Group 1 showed relatively lower level of quality of life than group 2 and group 3 in terms of the quality of life related to physical well-being. In addition, the control group reported higher quality of life in psychological health than both patients groups did. CONCLUSIONS: In conclusion, it is necessary for clinicians to make a careful evaluation of depressive tendency and eating disorders when obese women seek for medical help. The combination of medical treatment and psychological approach for obese women would result in higher quality of life. PMID- 17426434 TI - First steps towards molecular prevention in gastric carcinogenesis. PMID- 17426435 TI - Fermentation of starch and protein in the colon: implications for genomic instability. PMID- 17426436 TI - Come forth 1E10 anti-idiotype vaccine: delivering the promise to immunotherapy of small cell lung cancer. PMID- 17426437 TI - Loss of p53 and overexpression of EphA2 predict poor prognosis for ovarian cancer patients. PMID- 17426438 TI - Constitutive autophagy in plant root cells. AB - In previous studies, using a membrane-permeable protease inhibitor, E-64d, we showed that autophagy occurs constitutively in the root cells of barley and Arabidopsis. In the present study, a fusion protein composed of the autophagy related protein AtAtg8 and green fluorescent protein (GFP) was expressed in Arabidopsis to visualize autophagosomes. We first confirmed the presence of autophagosomes with GFP fluorescence in the root cells of seedlings grown on a nutrient-sufficient medium. The number of autophagosomes changed as the root cells grew and differentiated. In cells near the apical meristem, autophagosomes were scarcely found. However, a small but significant number of autophagosomes existed in the elongation zone. More autophagosomes were found in the differentiation zone where cell growth ceases but the cells start to form root hair. In addition, we confirmed that autophagy is activated under starvation conditions in Arabidopsis root cells. When the root tips were cultured in a sucrose-free medium, the number of autophagosomes increased in the elongation and differentiation zones, and a significant number of autophagosomes appeared in cells near the apical meristem. The results suggest that autophagy in plant root cells is involved not only in nutrient recycling under nutrient-limiting conditions but also in cell growth and root hair formation. PMID- 17426439 TI - A novel role for DNA mismatch repair and the autophagic processing of chemotherapy drugs in human tumor cells. AB - DNA Mismatch repair (MMR) maintains genome integrity by correcting DNA replication errors and blocking homologous recombination between divergent DNA sequences. The MMR system also activates both checkpoint and apoptotic responses following certain types of DNA damage. In a recent study, we describe a novel role for MMR in mediating an autophagic response to 6?thioguanine (6-TG), a DNA modifying chemical. Our results show that MMR proteins (MLH1 or MSH2) are required for signaling to the autophagic pathway after exposure to 6-TG. Using PFT-alpha, a p53 inhibitor, and shRNA-mediated silencing of p53 expression, we also show that p53 plays an essential role in the autophagic pathway downstream of the MMR system. This study suggests a novel function of MMR in mediating autophagy following chemical (6-TG) DNA mismatch damage through p53 activation. Here, we present the model and the clinical implications of the role of MMR in autophagy. PMID- 17426440 TI - A cycling protein complex required for selective autophagy. AB - Survival of environmental stress conditions requires the maintenance of cellular homeostasis. To preserve this balance, cells utilize a degradative mechanism known as autophagy. During this process, in response to starvation or other stresses, bulk cytoplasm is non-specifically sequestered within double-membrane vesicles and delivered to the lysosome/vacuole for subsequent degradation and recycling. The cytoplasm to vacuole targeting (Cvt) pathway is a type of specific autophagy, which occurs constitutively during growing conditions. Here, we examine three autophagy-related (Atg) proteins, Atg9, Atg23 and Atg27, which exhibit a unique localization pattern, residing both at the pre-autophagosomal structure (PAS) and other peripheral sites. These proteins colocalize, interact with one another in vivo, and form a functional complex. Furthermore, all three proteins cycle between the PAS and the other sites, and depend upon one another for this movement. Our data suggest that Atg9, Atg23 and Atg27 play a role in Atg protein retrieval from the PAS. In addition, Atg9 and Atg27 are the only known integral membrane Atg proteins involved in vesicle formation; a better understanding of their function may offer insight into the mechanism of membrane delivery to the PAS, the site of double-membrane vesicle assembly. PMID- 17426441 TI - LS14 cells: a model for chemoresistance in liposarcoma. AB - Liposarcoma, a malignancy of adipose tissue, is the most common soft tissue sarcoma. Patients whose primary tumor cannot be resected or those who have developed metastasis, have poor prognosis since liposarcomas are highly resistant to chemotherapy. We recently generated a spontaneously immortalized cell line, named LS14, from a patient with metastatic liposarcoma. Our goal was to compare the responsiveness of LS14 and SW872 liposarcoma cells to anti-cancer drugs and explore mechanisms of chemoresistance. Using complementary assays for cell viability and number we found that SW872 cells responded robustly to relatively low concentrations of doxorubicin, cisplatin and vinblastine. This reduction in cell viability was due to apoptosis, as evident by phosphatidylserine exposure and caspase 3 cleavage. In contrast, only a high dose of doxorubicin or combination therapy effectively reduced LS14 cell viability and induced apoptosis. LS14 cells showed a higher expression of Bcl-2 and Bcl-xL, but a lower expression of survivin and Bax, than SW872 cells, suggesting that anti-apoptotic proteins contribute to chemoresistance in LS14 cells. Although LS14 cells did not form colonies in soft agar, they generated large tumors and metastases in SCID mice, establishing their tumorigenicity in vivo. In conclusion, LS14 cells are much more resistant to chemotherapy than SW872 cells, making them an excellent model for exploring the efficacy and mechanism of action of anti-cancer drugs in liposarcomas. PMID- 17426442 TI - A new putative target for antisense gene therapy of glioma: glycogen synthase. AB - The treatment of malignant brain gliomas remains a challenge, despite the availability of the classical triad of surgery, radiotherapy, and chemotherapy. There is thus the need for investigations into other forms of treatment strategies, such as gene therapy. Using antisense technology we have targeted glycogen metabolism, since malignant astrocytes present a high content of glycogen. In vitro rat C6-glioma cells, transfected with antisense glycogen synthase (C6-AS cells) exhibited a decreased expression of glycogen synthase and reduced activity of glycogen synthesis, along with attenuated invasiveness. In vivo tumors induced by C6-AS cells in nude mice exhibited a significant reduction in tumor growth compared with controls. This reduction could be mediated by the induction of MCH-I expression. The inhibition of glycogen synthesis by antisense glycogen synthase validates a putative target and a new approach for further study to advance the much-needed efficacy of intervention strategies for malignant gliomas. PMID- 17426443 TI - Zebrafish tp53 knockdown extends the survival of irradiated zebrafish embryos more effectively than the p53 inhibitor pifithrin-alpha. AB - Cancer treatment strategies using radiation therapy have significantly improved outcomes in patients. Although advances in physics and electronics have enabled physicians to plan and deliver radiotherapy regimens precisely and efficiently, studies of the underlying molecular mechanisms of the radiation response could enable more effective and less toxic treatment strategies, as well as protection against radiation damage following unplanned exposure. Using the developing zebrafish embryo as a model system, we hypothesized that knockdown of tp53 mRNA or small molecule inhibition of p53 could alter the efficacy of radiotherapy against developing zebrafish embryos as a model for tumors. We found that interfering with p53 expression and function using hydroxylprolyl-phosphono peptide nucleic acid oligomers or pifithrin-alpha protected developing zebrafish embryos against radiation damage, consistent with our hypothesis. PMID- 17426444 TI - AKT1, AKT2 and AKT3-dependent cell survival is cell line-specific and knockdown of all three isoforms selectively induces apoptosis in 20 human tumor cell lines. AB - AKT is a key serine/threonine kinase in the PTEN/PI3K/AKT pathway(1) and activationof AKT is often observed in human cancers. To explore the role of AKT in cell survival in different tumor cells, we tested 20 human tumor cell lines for response to knockdown of AKT by small interference RNA (siRNA) and/or a kinase-dead mutant AKT. siRNA-mediated knockdown of all three AKT isoforms in tumor cell lines led to a reduction of phosphorylation of AKT substrates. Knockdown of AKT resulted in apoptosis in six out of 11 tumor cells with activated AKT. In contrast, knockdown of AKT induced apoptosis in three out of nine cell lines with a low level of active AKT. The responsiveness of the cells to knockdown of AKT was not affected by mutational status of p53 but appeared correlated with overexpression of HER2. To assess the role of individual AKT isoforms, five of the cell lines responsive to knockdown of AKT were further characterized. In ZR-75 cells, AKT1 is the predominant isoform responsible for cell proliferation and survival. Conversely, in IGROV1 cells, AKT2 plays a major role in cell proliferation, but no single isoform is essential for cell survival. Thus, the relative importance of the AKT isoforms is cell line-specific. Our data suggest that inhibiting all three AKT isoforms is necessary to elicit maximal apoptotic response in tumor cells, and the level of activated AKT is a favorable but not always reliable biomarker for preselection of responsive tumor cell lines to AKT inhibitors. PMID- 17426445 TI - A phase I clinical trial of an adenovirus-mediated endostatin gene (E10A) in patients with solid tumors. AB - PURPOSE: The purpose of the current study were to assess the safety and feasibility of repetitive intratumoral administration of E10A, an adenoviral vector encoding the wild-type endostatin gene, to patients with solid tumors, and to evaluate its biologic effect and the pharmacokinetics of endostatin. METHODS: Patients were treated with escalating doses from 1 x 10(10) VP to 1 x 10(12) VP of E10A intratumorally on days 1 and 8. Patients were assessed for toxicity and viral shedding, and antitumor response was evaluated by imaging techniques and tumor biopsy. Circulating levels of endostatin were examined. RESULTS: Fifteen patients received 29 injections of E10A. No dose-limiting toxicity was developed, and the maximum tolerated dose had not yet been reached. Fever and local reaction of injection site were common, but rarely severe. Mild and transient hepatotoxicity was observed in one patient. Minor response of injected tumor was achieved and improvement of the control tumor was observed in one patient with nasopharyngeal carcinoma, and tumor necrosis was occurred in two patients. Sustained elevation of serum endostatin levels was detected. CONCLUSION: Weekly intratumoral injection of up to 1 x 10(12) VP of E10A to patients with solid tumor is a feasible and well-tolerated procedure that exerts mild antitumor effects. A small and sustained elevation of endogenous endostatin in blood possibly has antitumor activity. PMID- 17426446 TI - Phi29 pRNA vector for efficient escort of hammerhead ribozyme targeting survivin in multiple cancer cells. AB - Ribozymes are potential therapeutic agents which suppress specific genes in disease-affected cells. Ribozymes have high substrate cleavage efficiency, yet their medical application has been hindered by RNA degradation, aberrant cell trafficking, or misfolding when fused to a carrier. In this study, we constructed a chimeric ribozyme escorted by the motor pRNA of bacteriophage phi29 to achieve proper folding and enhanced stability. A pRNA molecule contains an interlocking loop domain and a 5'/3' helical domain, which fold independently of one another. When a ribozyme is connected to the helical domain, the chimeric pRNA/ribozyme reorganizes into a circularly permuted form, and the 5'/3' ends are relocated and buried in the original 71'/75' positions. Effective silencing of the anti apoptotic gene survivin by an appropriately designed chimeric ribozyme, as demonstrated at mRNA and protein levels, led to programmed cell death in various human cancer cell lines, including breast, prostate, cervical, nasopharyngeal, and lung, without causing significant non-specific cytotoxicity. Through the interlocking interaction of right and left loops, monomer pRNA/ribozyme chimeras can be incorporated into multi-functional dimer, trimer and hexamer complexes for specific gene delivery. Using the phi29 motor pRNA as an escort may revive the ribozyme's strength in medical application. PMID- 17426447 TI - Caspase-8 preferentially senses the apoptosis-inducing action of NG-18, a Gambogic acid derivative, in human leukemia HL-60 cells. AB - Gambogic acid (GA) is the major active ingredient of gamboge secreted from a Chinese traditional medicine Garcinia hanburryi possessing potent anti-tumor activity. N-(2-ethoxyethyl)gambogamide (NG-18), a derivative of GA, also efficiently inhibits proliferation of cultured human tumor cells. The inhibition effect of NG-18 is associated with its ability to induce apoptosis. In the present study, NG-18 markedly induced leukemia HL-60 cells apoptosis, and the extrinsic and intrinsic apoptosis pathways were activated almost at the same time. NG-18-induced tumor cell apoptosis was associated with up-regulation of pro apoptotic Bcl-2 family member Bax, and downregulation of anti-apoptotic protein Bcl-2. The NG-18-induced apoptosis was blocked completely by a pan-caspase inhibitor Z-VAD-FMK, indicating that caspases were functionally and actively involved in this process. The specific inhibition of caspase-8 activity using Z IETD-FMK significantly blocked NG-18-induced apoptosis. In contrast, inhibition of other initiator caspases, caspase-2 or -9, using Z-VDVAD-FMK or Z-LEHD-FMK respectively had no effect on NG-18-induced apoptosis. Altogether, our data demonstrated that NG-18-induced apoptosis was dependent on caspases and caspase-8 acted as a key executor in the event. PMID- 17426448 TI - A novel polypeptide from shark cartilage with potent anti-angiogenic activity. AB - Using guanidine-HCl extraction, acetone precipitation, ultra-filtration and chromatography, a novel polypeptide with potent anti-angiogenic activity was purified from cartilage of the shark, Prionace glauca. N-terminal amino acid sequence analysis and SDS-PAGE revealed that the substance is a novel polypeptide with MW 15500 (PG155). The anti-angiogenic effects of PG155 were evaluated using zebrafish embryos model in vivo. Treatment of the embryos with 20 microg/ml PG155 resulted in a significant reduction in the growth of subintestinal vessels (SIVs). A higher dose resulted in almost complete inhibition of SIV growth, as observed by endogenous alkaline phosphatase (EAP) staining assay. An in vitro transwell experiment revealed that the polypeptide inhibited vascular endothelial growth factor (VEGF) induced migration and tubulogenesis of human umbilical vein endothelial cells (HUVECs). Exposure of HUVECs in 20 microg/ml PG155 significantly decreased the density of migrated cells. Almost complete inhibition of cell migration was found when HUVECs were treated with 40-80 microg/ml PG155. PG155 (20 microg/ml) markedly inhibited the tube formation of HUVECs and a dose dependent effect was also found when treatment of HUVECs with PG155 at the concentration from 20-160 microg/ml. PMID- 17426449 TI - Cell cycle dependent nucleolar localization of the coronavirus nucleocapsid protein. AB - The nucleolus is a dynamic sub-nuclear structure which is involved in ribosome subunit biogenesis, modulation of cell growth and response to cell stress. The nucleolar proteome varies particularly with regard to the cell cycle. Viral proteins can localise to the nucleolus and using the coronavirus nucleocapsid (N) protein as a model, the cell cycle dependent trafficking of viral proteins to the nucleolus was investigated. Cell synchronisation studies coupled to live cell confocal microscopy indicated that nucleolar localisation of N protein was greater in the G2/M phase of the cell cycle than at other stages. This result was reinforced when FRAP and FLIP analysis indicated that N protein was more mobile within the nucleoplasm and nucleolus in the G2/M phase of the cell cycle. The data suggested that viral nucleolar proteins can also localise to the nucleolus in a cell cycle dependent manner and this may be related to dynamic trafficking. PMID- 17426450 TI - R-spondins in cutaneous biology: nails and cancer. AB - The R-spondins are a relatively recently identified family of secreted proteins linked to the Wnt signaling pathway. Recently, human phenotypes have been associated with mutations in two human R-spondins. Germline RSPO4 and RSPO1 mutations result in recessive anonychia (absence of all fingernails and toenails) and a recessive syndrome with XX-male sex reversal, PPK and a predisposition to SCC, respectively. This review discusses the key roles R-spondins play in embryogenesis, adult tissue maintenance and skin carcinogenesis. PMID- 17426451 TI - p27Kip1 metabolism: a fascinating labyrinth. AB - The progression through the phases of cell division cycle is regulated by different cyclins and cyclin-dependent kinases (CDKs) complexes. Due to their key function, the activity of cyclin/CDK complexes is controlled by several mechanisms, including the inhibition by a number of proteins collectively defined CDK inhibitors or CKIs. Among the CKIs, p27Kip1 represents a protein of central activity for the control of several phenotypes, including proliferation, differentiation and malignant transformation. p27Kip1 belongs to the growing family of "natively unfolded," "intrinsically disordered" or "intrinsically unstructured" proteins. The disorder proteins present a very large number of possible conformations that, after the binding, converge to a well-defined structure with an extraordinary affinity for the target. As matter of fact, the absence of a pre-existing folding strongly facilitates p27Kip1 interaction with a number of targets. Until recently, p27Kip1 has been solely viewed as a nuclear protein with the function of modulating cyclin-CDK activity and hence, cell cycle progression. However, exhaustive studies have now demonstrated that the protein plays additional roles outside of the nucleus, including, particularly, the control of cell motility. Thus, the cellular localization is of fundamental importance in p27Kip1 function. Accordingly, at least two different mechanisms of degradation, occurring either in the nucleus or in the cytosol, have been observed. Convincing evidences have demonstrated that p27Kip1 is a phosphoprotein showing at least six to eight phosphorylatable residues. However, the precise functional roles of the phosphorylations and the identification of the kinases responsible for the post-synthetic modifications are still debated. In this brief review, we will report the Literature data that connect the post-synthetic modifications of p27Kip1 with its function, localization and metabolism. The picture that emerges demonstrates that several of the pieces of the CKI metabolism are still nebulous. PMID- 17426452 TI - HTRA1 variant confers similar risks to geographic atrophy and neovascular age related macular degeneration. AB - Age-related macular degeneration (AMD) is the most common cause of irreversible visual impairment in the developed world. The two forms of advanced AMD, geographic atrophy (GA) and choroidal neovascularization (wet AMD), represent two types of degenerative processes in the macula that lead to loss of central vision. Soft confluent drusen, characterized by deposits in macula without visual loss are considered a precursor of advanced AMD. A single nucleotide polymorphism, rs11200638, in the promoter of HTRA1 has been shown to increases the risk for wet AMD. However, its impact on soft confluent drusen and GA or the relationship between them is unclear. To better understand the role the HTRA1 polymorphism plays in AMD subtypes, we genotyped an expanded Utah population with 658 patients having advanced AMD or soft confluent drusen and 294 normal controls and found that the rs11200638 was significantly associated with GA. This association remains significant conditional on LOC387715 rs10490924. In addition, rs11200638 was significantly associated with soft confluent drusen, which are strongly immunolabeled with HTRA1 antibody in an AMD eye with GA similar to wet AMD. Two-locus analyses were performed for CFH Y402H variant at 1q31 and the HTRA1 polymorphism. Together CFH and HTRA1 risk variants increase the odds of having AMD by more than 40 times. These findings expand the role of HTRA1 in AMD. Understanding the underlying molecular mechanism will provide an important insight in pathogenesis of AMD. PMID- 17426453 TI - p63: the phantom of the tumor suppressor. AB - The last twenty years of research into p53 function has revealed some fascinating discoveries into the orchestration of tumor suppressor pathways with a multitude of putative drug targets being investigated. However, it was not until 1998 that the ancestral mother of p53 was documented. The eldest evolutionary conserved homolog of the p53 family is known today as p63. Originally, it was thought p63 was another tumor suppressor that could function in a similar capacity to p53. However, elegant demonstrations of the divergent roles that p63 plays as a key transcriptional regulator of the proliferation and differentiation cascade in stratified epithelia are documented. These data link deltaNp63alpha to adult tissue stem cell regulation and possibly "cancer stem cells". p63 lacks mutation in cancer development, which is in stark contrast to the classically high mutation status of p53 in a large compendium of cancer types. Perhaps suggesting a selective preference for p53 mutation. Why is p63 rarely mutated despite being part of the same gene family? Interestingly, p63 is often over-expressed and amplified in cancer, thus revealing a paradox. Is p63 required to provide cancer cell populations with a selective advantage as much as a loss of p53 function by mutation? Has p53 been masking a "phantom" with promising features as a target for drug development? Can we exploit the biochemical know how gained from the mass of p53 research to further elucidate deltaNp63alpha gene function? In this review, we will summarise the emerging advances that are elucidating deltaNp63alpha as a promising drug target. PMID- 17426454 TI - NFkappaB-independent signaling to the cyclin D1 gene by Rac. AB - In this report we characterize the mechanism of Rac-mediated cyclin D1 gene expression in mouse embryonic fibroblasts. Activated Rac strongly stimulated cyclin D1 gene transcription but did not alter the half-life of cyclin D1 mRNA. Inhibition of NFkappaB signaling with the IkappaB super-repressor blocked the Rac dependent expression of cyclin D1 mRNA, and this effect was selective since ERK dependent cyclin D1 mRNA induction was minimally affected by super-repressor expression. However, we found that p65 activity in this system was induced by serum and not by activated Rac. Moreover, mouse cyclin D1 promoter-luciferase assays showed that Rac stimulated cyclin D1 gene expression without activating NFkappaB and that an essential Rac-regulated promoter element is located far upstream or downstream of the cyclin D1 transcription start site. We conclude that, in MEFs, Rac-mediated induction of cyclin D1 mRNA requires activation of a parallel NFkappaB pathway whereas ERK induces cyclin D1 transcription independent of NFkappaB. PMID- 17426455 TI - Macrophage migration inhibitory factor coordinates DNA damage response with the proteasomal control of the cell cycle. AB - Proper repair of DNA damage is critical for protecting genomic stability, cellular viability and suppression of tumorigenesis. Both p53-dependent and p53 independent pathways have evolved to coordinate the cellular response following DNA damage. In this review, we highlight the importance of the ubiquitously expressed protein macrophage migration inhibitory factor (MIF) for an appropriate response to DNA damage. We discuss the mechanisms by which MIF affects the activity of the ubiquitin-proteasome system, and how this impacts on the integrity of the genome and on cancer. PMID- 17426456 TI - The ever expanding role for c-Myc in promoting genomic instability. AB - Genomic instability (GI) is a hallmark of many cancers. GI is believed to confer upon impending neoplastic cells the ability to accumulate all of the requisite mutations for transformation within the relatively short time-frame of an organism's lifespan. Recently described properties of the c-Myc oncoprotein show that, in addition to its directly transforming role, it can mediate GI via the induction of reactive oxygen species and by promoting whole chromosome instability leading to tetraploidy and aneuploidy. Mediators of both properties have been identified and have begun to provide a framework within which to understand not only how c-Myc alters the genome but how it might also cooperate with its more traditional transforming activities. These genome-altering properties of c-Myc suggest that they provide the protein with the ability to confer a "mutator phenotype" to cells in which its expression is deregulated. PMID- 17426458 TI - Obesity and the risk of colon polyps. PMID- 17426457 TI - High-throughput screening of stabilizers for respiratory syncytial virus: identification of stabilizers and their effects on the conformational thermostability of viral particles. AB - Respiratory syncytial virus (RSV) is the leading cause of severe respiratory infection in children worldwide. Recombinant live attenuated viral preparations are one of the most promising strategies for vaccination but they typically possess poor thermostability. In this work, a library of compounds was screened and stabilizers were selected based on their ability to inhibit the aggregation of RSV perturbed at 56 degrees C. After screening and selection of excipients, the conformational stability of the RSV proteins was evaluated in the presence of potential stabilizers. The secondary and tertiary structures as well as aggregation/dissociation of RSV were monitored using circular dichroism and second derivative UV absorption spectroscopies and light scattering, respectively, as a function of temperature (10-90 degrees C). RSV membrane fluidity was also evaluated by generalized polarization of Laurdan fluorescence. Screening experiments showed that a variety of sugars, amino acids, polyols and polyanions inhibited the aggregation of viral particles. Conformational stability studies demonstrated that the addition of sugars and polyols stabilized RSV as indicated by a significant increase in the transition melting temperature (Tm) of both the secondary and tertiary structures as well as the gel to liquid crystalline membrane transition. These results should provide the basis for rational development of more physically stable formulations of live attenuated RSV vaccines. PMID- 17426459 TI - Desmoid tumors and familial adenomatous polyposis: a tale of two syndromes. PMID- 17426460 TI - GERD evaluation: time for a new paradigm? AB - In the past there has been reliance on heartburn and acid regurgitation as the cardinal symptoms of gastro-esophageal reflux. Now it is recognized that diverse additional symptoms including extraesophageal manifestations are essential components of gastroesophageal reflux disease (GERD). Furthermore, previous reliance in mucosal damage at endoscopy as evidence of therapeutic success in the management of GERD has proven of only limited value in the diverse world of GERD symptoms. Indeed is now apparent that the relationship between symptoms and lesions is modest at best. Most patients with GERD have no overt erosions or ulceration at endoscopy and they frequently suffer from a wide array of complaints other than heartburn that contribute to their diminished perception of life's quality. To address these problems, multidimensional questionnaires designed to encompass the diverse symptomatology of GERD have been investigated, principally, to provide a reproducible metric of change in the subjective response to pharmacologic therapy during trials for patients with nonerosive reflux disease. The most promising of these instruments is The ReQuest questionnaire, because it has been extensively validated in multiple patient groups and cultures, appears highly promising in this regard and for erosive disease too and may ultimately prove to be of value for the initial evaluation of patients with GERD. We propose that accurate symptom delineation complemented with endoscopy performed in selected groups of patients will become the mainstay in diagnosis of GERD and the assessment of therapy efficacy. PMID- 17426461 TI - Obscure gastrointestinal bleeding: diagnostic and therapeutic approach. AB - Obscure gastrointestinal bleeding (OGIB) is defined as an intermittent or continuous loss of blood in which the source has not been identified after upper endoscopy and colonoscopy. It constitutes a diagnostic and therapeutic challenge for the general internist and the gastroenterologist. This article provides an overview of the etiology, clinical presentation, and diagnostic modalities of OGIB including push enteroscopy, double balloon enteroscopy, wireless capsule endoscopy, enteroclysis, angiography, bleeding scanning with labeled red blood cells, and surgery with intraoperative enteroscopy. Therapeutic modalities including iron replacement, combined hormones, octreotide acetate, therapeutic endoscopy, and surgery are also discussed. In addition, a rational approach to patients with OGIB according to the clinical presentation is presented herein. PMID- 17426462 TI - Distinguishing eosinophilic esophagitis in pediatric patients: clinical, endoscopic, and histologic features of an emerging disorder. AB - GOALS: To determine the clinical, endoscopic, and histologic criteria that distinguish children with eosinophilic esophagitis (EE) from those with non-EE diagnoses. BACKGROUND: EE is a disease of escalating incidence. Distinguishing children with EE from those with non-EE diagnosis can be difficult before endoscopy. STUDY: A retrospective case-control study was performed for children with any degree of esophageal eosinophilic inflammation who underwent esophageal biopsy at Children's Hospital San Diego from January 1998 to December 2002. A database containing children who met histologic criteria for EE and an equivalent number of children who had milder esophageal eosinophilia (non-EE patients) was created to compare the 2 groups. RESULTS: The number of EE cases increased from 15 in 1998 to 35 in 2002. EE patients were predominantly school-aged boys; 5 of 102 were suspected to have EE before biopsy. Although EE and non-EE patients complained of vomiting and abdominal pain at equivalent rates, EE patients were 3 times more likely to complain of dysphagia [odds ratio (OR)=3.11, 95% confidence interval (CI) 1.55-6.65] and twice as likely to have stricture formation (OR=2.43, 95% CI 0.72-11.75). On endoscopy, patients with EE were 19-times more likely than non-EE patients to have endoscopic abnormalities (OR=19, 95% CI 9.0 45.88). Histologically, EE patients were more likely to have basal zone hyperplasia and degranulated eosinophils (OR=45 and 157, respectively). CONCLUSIONS: We demonstrate that school-aged children, particularly boys, who complain of dysphagia should raise the index of suspicion for EE. We also suggest that EE-associated strictures are more common than peptic strictures in children. PMID- 17426463 TI - Esophagitis in frail elderly people. AB - INTRODUCTION: We studied the clinical course of elderly patients acutely hospitalized for various diseases, assessing any differences between patients with and without esophagitis. STUDY: A case-control study on the presence of esophagitis was conducted on the clinical records of all in-patients undergoing gastroduodenoscopy at Padova Geriatric Hospital from 1997 to 2001. Data were examined on 338 sex-matched patients: 169 with a diagnosis of esophagitis and 169 with a negative endoscopy. RESULTS: Admissions for acute respiratory disorders [odds ratios (OR) 2.68; 95% confidence interval (CI) 0.89-8.01], a remote diagnosis of esophagitis (OR 11.34; 95%CI 2.68-48.07), obesity (OR 3.36; 95%CI 0.91-12.48), and being bedridden (OR 6.84; 95%CI 3.27-14.29) were found to be independent risk factors for the presence of esophagitis. The symptoms prompting the endoscopic diagnoses included: gastrointestinal bleeding (OR 7.61; 95%CI 2.76 21.0), heartburn (OR 4.58; 95%CI 1.86-11.28), and cough (OR 3.59; 95%CI 1.34 9.62). Steroids (OR 2.68; 95%CI 1.11-6.44) and calcium antagonists (OR 1.50; 95%CI 0.79-2.87) were associated with esophagitis as risk factors, whereas proton pump inhibitors (OR 0.46; 95%CI 0.25-0.87), nitrates (OR 0.14; 95%CI 0.02-0.78), and sucralfate in males (OR 0.09; 95%CI 0.01-0.92) were associated as protective factors. Patients with esophagitis were discharged with an endocrinologic/metabolic-type diagnosis. Deaths were significantly higher among patients with esophagitis (25 vs. 9); more severe esophagitis was characterized by a higher Charlson comorbidity index and a greater presence of anorexia and nausea. CONCLUSIONS: These findings seem to substantiate the theory that esophagitis is a characteristic which exacerbates frailty in hospitalized elderly people and its identification may be helpful in these patients. PMID- 17426464 TI - Functional chest pain: esophageal or overlapping functional disorder. AB - BACKGROUND: Whether patients with functional chest pain have an esophageal or overlapping functional disorder of the gut is unclear. We investigated the prevalence of functional gastrointestinal disorders in patients with functional chest pain. METHODS: One hundred patients with functional chest pain and normal cardiac, endoscopic, and manometric studies were evaluated for esophageal hypersensitivity with a balloon distension test. Subsequently, a modified Rome II functional bowel disorder questionnaire was mailed to these subjects. Prevalence of irritable bowel syndrome (IBS) and other functional disorder were determined using the Rome II criteria. In addition, we assessed the prevalence of chest pain in 81 patients with functional constipation. RESULTS: There were 69 responders (54 women); 2 were excluded. Fifty-five patients (82%) fulfilled criteria for other functional disorders besides chest pain. Although there was an overlap, IBS (27%) and abdominal bloating (22%) were most common; dyspepsia (7%), dysphagia (7%), nonspecific bowel disorder (7%), constipation (4%), abdominal pain (3%), and diarrhea (1%) were less common. Among responders, 52 (78%) had esophageal hypersensitivity and 15 (22%) had normosensitivity, with similar prevalence of functional disorders. Thirty-two (39%) of the subjects with functional constipation reported chest pain occasionally, and 5 (6%) frequently. CONCLUSIONS: Approximately 80% of patients with functional chest pain exhibit features of other functional disorders including IBS suggesting an overlap. This association is independent of esophageal hypersensitivity. Recognition of this overlap may facilitate better management of these patients. PMID- 17426465 TI - Therapeutic effect of melatonin in patients with functional dyspepsia. AB - BACKGROUND AND GOAL: Melatonin may inhibit reactive oxygen species-related pathogenesis in the alimentary tract by neutralizing free radicals. In the present study we assessed the potential protective action of melatonin in ulcerlike dyspepsia. STUDY: Sixty patients aged 19 to 39 years with the diagnosis of functional dyspepsia according to the Rome Criteria II and no Helicobacter pylori infection were involved in the study. Melatonin, at a dose of 5 mg (n=30), or placebo (n=30) were taken in the evening for a period of 12 weeks. At this time, patients were on an equivalent diet and were only to take an alkaline drug in case of the abdominal pain. RESULTS: After 12 weeks, the dyspeptic symptoms completely subsided in 17 patients in the melatonin-treatment group (56.6%). In other 9 individuals (30.0%) a partial improvement in health was achieved, especially in the frequency and intensity of nocturnal pain. After placebo, the majority of patients (93.3%) did not experience any improvement in symptoms. Multivariate analysis indicated that melatonin (odds ratio 95.86, 95% confidence interval 3.72-2469.37, P<0.01) correlated independently with significantly improved patients health. H. pylori past infection decreased positive effect of melatonin in ulcerlike dyspepsia. CONCLUSIONS: Melatonin can be considered as an auxiliary drug in the treatment of ulcerlike dyspepsia. PMID- 17426466 TI - Is TCRgamma clonality assay useful to detect early celiac disease? AB - BACKGROUND: Although histology is considered the gold standard for the diagnosis of celiac disease, the early stages (latent or potential) of this disease are difficult to diagnose, because of the negativity of laboratory tests and the lack of villous atrophy. Thus, markers of early disease are needed. AIMS: We investigated the possibility to detect latent or potential celiac disease by means of TCRgamma clonality assay in intraepithelial T cells in patients with suspected disease, negative laboratory tests, and an increased number of intraepithelial lymphocytes. PATIENTS AND METHODS: Duodenal biopsies were obtained from 35 patients with nonspecific duodenitis (controls), 13 latent or potential celiac disease subjects, 28 well-defined celiac patients, and 8 celiac patients in gluten-free diet. Histologic and immunohistochemical quantification of intraepithelial lymphocytes, as well as TCRgamma clonality assay, were carried out in all subjects by means of standard techniques. RESULTS: Intraepithelial lymphocytes and TCRgamma clonality were significantly increased in potential and defined celiac patients with respect to the controls, even though the increase in TCRgamma clonality was lesser with respect to that of intraepithelial lymphocytes. No significant differences were found concerning this variable between the potential and defined celiac subjects. CONCLUSIONS: TCRgamma clonality does not represent a marker of early disease. However, it might be useful to help in distinguishing celiac disease from other causes of nonspecific duodenitis. PMID- 17426467 TI - Risk factors for ulcerative colitis in a Chinese population: an age-matched and sex-matched case-control study. AB - BACKGROUND AND AIMS: Cigarette smoking, alcohol use, appendectomy, and family history of inflammatory bowel disease (IBD) have all been shown to be associated with IBD, but there were no reports of risk factors for IBD in a Chinese population in which the incidence of IBD is increasing during the past decade. We conducted a case-control study to examine associations between previously reported environmental risk factors and development of ulcerative colitis (UC) in Wuhan city, central China. METHODS: A total of 177 patients with UC and 177 age matched and sex-matched controls were prospectively studied in Wuhan city from January 2004 to December 2004. An age-matched and sex-matched case-control study was conducted to assess the role of smoking, alcohol use, appendectomy, and other potential risk factors in the development of UC by a detailed questionnaire. RESULTS: Smoking was a protective factor and exsmoking is a risk factor for UC [compared with nonsmokers, smokers: odds ratios (OR)=0.28, 95% confidence intervals (CI): 0.16-0.48, P=0.0001; exsmokers: OR=4.36, 95%CI: 1.46-13.04, P=0.008]. Positive family history of IBD was a risk factor (OR=4.35, 95%CI: 1.21 15.71, P=0.025) whereas appendectomy was a protective factor (OR=0.24, 95%CI: 0.07-0.86, P=0.028) for UC. There were no significant associations between UC and other factors examined. CONCLUSIONS: Although the incidence of UC in Chinese is relatively lower than that in white, the same risk factors for UC that were reported in white populations were associated with Chinese UC patients. Specifically, smoking was a protective factor for UC and exsmoking was associated with an increase risk of UC in a Chinese population. Family history of IBD was shown to be a risk for UC, whereas appendectomy was associated with a low risk for UC. PMID- 17426468 TI - Body mass index: a marker for significant colorectal neoplasia in a screening population. AB - BACKGROUND AND AIMS: Although some studies suggest a positive association between increasing body mass index (BMI) and risk for colorectal neoplasia, the impact on screening has not been examined. We performed a cross-sectional study to examine the association of BMI and colorectal neoplasia in a screening population. METHODS: Data collected for 2493 patients presenting for screening colonoscopy included known risk factors for colorectal neoplasia, demographic information, and lifestyle factors. Our outcome was the endoscopic detection of significant colorectal neoplasia which included adenocarcinoma, high-grade dysplasia, villous tissue, adenomas 1 cm or greater and multiple (>2) adenomas of any size. RESULTS: Overall, we observed an increased risk and prevalence for significant colorectal neoplasia in women as BMI increased (P value for trend <0.002). This relationship was the strongest for the women with a BMI > or =40 (odds ratios=4.26; 95% confidence intervals=2.00-9.11). There was no such relationship in our male population. CONCLUSIONS: Increasing BMI, in our population, was associated with an increase risk for colorectal neoplasia in female patients. This study reinforces the importance of screening colonoscopy especially in obese women. PMID- 17426469 TI - Risk of colorectal cancer in women with a prior diagnosis of gynecologic malignancy. AB - GOALS AND BACKGROUND: Earlier studies regarding the risk of colorectal cancer (CRC) in women with a prior diagnosis of gynecologic malignancies have revealed conflicting results. We sought to further clarify this association. METHODS: A retrospective cohort study was performed using the General Practice Research Database of the United Kingdom. Patients with a prior diagnosis of ovarian, uterine, or cervical cancers were compared with control patients without a prior gynecologic malignancy. The primary outcome was a diagnosis of CRC. Poisson regression analysis was used to assess the effects of potential confounders. RESULTS: The study included 1995 ovarian, 1348 uterine, and 1101 cervical cancer patients and 7980, 5392, and 4404 matched control patients, respectively. The adjusted incidence rate ratio (IRR) of CRC among ovarian cancer patients was 2.90 [95% confidence intervals (CI) 1.45-5.82]. Five of 10 cases of CRC in ovarian cancer patients were diagnosed within 6 months of the cancer diagnosis with an adjusted IRR of 8.0 (95% CI 1.9-33.6). Excluding the initial 6 months of follow up after the diagnosis of ovarian cancer, the adjusted IRR was 1.6 (95% CI 0.76 5.03). The adjusted IRR of CRC in patients with a prior diagnosis of uterine and cervical cancer was 0.79 (95% CI 0.24-2.61) and 1.50 (95% CI 0.43-5.21), respectively. CONCLUSIONS: Women with a prior diagnosis of ovarian cancer are at an increased risk of CRC. The risk of CRC was not increased among patients with a prior history of uterine and cervical cancer. PMID- 17426470 TI - 3' Mutation of the APC gene and family history of FAP in a patient with apparently sporadic desmoid tumors. AB - Desmoid tumors may occur sporadically or as part of the extraintestinal manifestations of familial adenomatous polyposis. Different phenotypes have been described and some genotype-phenotype correlations have been raised, associated with different sites of germline mutations in the adenomatous polyposis coli (APC) gene. We report on a 42-year-old woman ascertained for a large desmoid tumor of the anterior chest wall with pleural involvement, which persistently recurred despite a decade of treatment including hormone therapy, chemotherapy, and surgery. Spontaneous disappearance of the tumor was later noted after 1 year without any treatment and confirmed after 4 years of regular follow-up. Repeated colonoscopies were normal in the proband and DNA sequencing showed a frameshift mutation due to a single adenosine deletion at position 5772 (codon 1924). This mutation, located in the exon 15 at the 3' end of the APC gene, leads to an unusual and late onset phenotype. The pedigree revealed other isolated or familial adenomatous polyposis-associated cases of desmoid tumors. This family report shows that a molecular analysis of the APC gene should be performed in familial desmoid tumors for accurate genetic counseling and follow-up. PMID- 17426471 TI - Bile leak after laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy is commonly performed as the treatment of choice for symptomatic gallstone diseases. Bile leak is a potential complication of this procedure and the cystic duct stump is the most common site of leakage. Early diagnosis and treatment of bile leak is crucial in decreasing the morbidity and mortality related to this complication. Endoscopic retrograde cholangiopancreatography with stent placement and/or sphincterotomy is highly effective in the diagnosis and treatment of this problem. PMID- 17426472 TI - Factors associated with abnormal gastric emptying in alcohol-related chronic pancreatitis. AB - BACKGROUND: Patients with alcohol-related chronic pancreatitis (ARCP) may present with abnormal gastric emptying (GE), which has been ascribed mainly to nutrient maldigestion. Nevertheless, many patients also have diabetes with autonomic dysfunction and malnutrition and the role of these factors on abnormal GE has not been investigated. GOALS: To determine the influences of malabsorption, diabetes, malnutrition, and autonomic dysfunction on GE abnormalities in patients with ARCP. STUDY: Forty ARCP outpatients and 18 healthy controls were studied. GE was measured by scintigraphy after a standard, liquid, nutrient meal labeled with Technetium-phytate. Autonomic function was evaluated by cardiovascular tests. The influence of each factor on abnormal GE was assessed by Bayesian multiple regression analysis. RESULTS: In the ARCP group, GE was abnormal in 19 patients (47.5%), who showed either accelerated (N=12) or delayed emptying (N=7). Diabetes was highly prevalent (P<0.01) in ARCP patients with either rapid or delayed GE (18/19). Multiple regression analysis showed that not only diabetes, but also autonomic dysfunction has significant effects on abnormal GE, whereas malabsorption and malnutrition seemed not to be associated to abnormal emptying. CONCLUSIONS: A substantial proportion of patients with ARCP may have abnormal GE. Either delayed or accelerated GE seem to be related to underlying diabetes mellitus and autonomic neuropathy rather than to nutrient malabsorption and malnutrition. PMID- 17426473 TI - Efficacy of preoperative albendazole use according to months in hydatid cyst of the liver. AB - PURPOSE: Hydatid cyst is still a significant health and economical problem due to insufficient preventive measures. Infestation by hydatid disease in humans most commonly occurs in the liver and causes complications unless treated properly. Therefore, hepatic hydatid cyst should immediately be treated when diagnosed. Today, the treatment of hydatid cysts is principally surgical. To avoid recurrence, viable scoleces in the hydatid cyst should be eliminated before emptying surgically. Length of Albendazole therapy, a drug effective on scoleces in the cyst, is still controversial. METHODS: In this study, there was a significant difference in the rate of viable scoleces when group III receiving albendazole for 3 months preoperatively was compared with group I, group II, and the control group. We do think 3-month preoperative abendazole treatment is effective in reducing the rate of recurrence and if still viable scoleces are observed in the surgery, albendazole treatment should be continued for 1 month postoperatively. RESULTS: After the therapy, in group I 10 (47.6%), in group II 7 (33.3%), and in group III 2 (0.9%) patient's scoleces were alive. In the control group (group IV) 17 (80%) patients' scoleces were intact. When group III is compared with the control group, a significant difference was observed (P<0.05). Yet again, there was a significant difference between the groups when groups I and II were compared with group III. CONCLUSIONS: We concluded that in treatment of hydatid cyst, albendazole should be continued at least for 3 months preoperatively and if still viable scoleces are identified, medical treatment should follow the surgical intervention at least for 1 month to decrease the possibility of residual cysts and recurrence. PMID- 17426474 TI - The clinical characteristics of rhabdomyolysis in patients with liver cirrhosis. AB - BACKGROUND: We have frequently seen idiopathic rhabdomyolysis developed after liver cirrhosis (LC) in our hospital. This has rarely been reported, and no studies have compared it with other cases of rhabdomyolysis. GOALS: We attempted to examine the pathologic characteristics of rhabdomyolysis development and to assess the prognosis in patients with LC. STUDY: We retrospectively reviewed the medical records of 243 patients (261 cases) with rhabdomyolysis and selected 74 patients (91 cases) for the study group, who concurrently had LC. Seventy-five patients (76 cases) with no evidence of LC were served as controls. RESULTS: In 59.3% of the LC-group patients, the causes of rhabdomyolysis were not identified. In 10.5% of the control-group patients, the causes of rhabdomyolysis proved to be unknown (P=0.000). In 17.6% of the LC-group patients, 30 cases (33.0%) of the recurrent rhabdomyolysis were identified. In the control group, only one case of the recurrent rhabdomyolysis was noted (P=0.001). The mortality was significantly higher in the LC group than the control group (27.5% vs. 14.5%) (P=0.042). Of 25 LC-group patients who died, 96.0% had Child-Pugh classification C. In addition, 84.0% developed acute renal failure during the course. Coexistent infection, hepatic encephalopathy, and the elevated levels of serum lactate dehydrogenase and C-reactive protein were also important prognostic factors. CONCLUSIONS: In conclusion, rhabdomyolysis is developed without specific causes in patients with LC, and it is serious and often fatal particularly in cases in which acute renal failure and severe hepatic dysfunction exist. Our results indicate that LC is the underlying disease for the development of rhabdomyolysis. PMID- 17426475 TI - Psychiatric comorbidity does not predict interferon treatment completion rates in hepatitis C seropositive veterans. AB - GOALS: The aim of this study was to evaluate the impact of common psychiatric disorders on treatment completion of antiviral therapy prescribed to a series of hepatitis C virus (HCV) positive US veterans. BACKGROUND: Clinical experience suggests that preexisting psychiatric conditions may adversely affect the ability to tolerate combination antiviral therapy in patients with HCV infection. STUDY: We performed a retrospective chart review of 130 HCV positive veterans treated with combination antiviral therapy [interferon (IFN)/ribavirin] at VA San Diego from 2000 to 2004. We examined baseline psychiatric and substance use diagnoses, as well as demographic and comorbid medical disease variables for all patients started on treatment. RESULTS: Thirteen percent of patients in our cohort required treatment discontinuation for neuropsychiatric adverse effects. There was no association between treatment completion and any specific psychiatric diagnosis, baseline use of antidepressants, history of substance abuse/dependence, or combined psychiatric and substance use diagnoses for patient groups receiving either standard or pegylated IFN plus ribavirin therapies. Psychiatric and substance use disorders were not associated with dropout due to neuropsychiatric adverse effects. Baseline comorbid medical disorders also did not predict treatment completion. However, higher body weight did predict likelihood of treatment completion, especially for those > or =100 kg compared with thinner subjects (odds ratio=2.90; P=0.037). CONCLUSIONS: In this cohort of veterans, prior psychiatric or substance use history did not predict completion of recommended IFN/ribavirin treatment. These findings suggest that a larger pool of veterans with psychiatric or substance use disorders may be considered candidates for antiviral therapy when provided with multidisciplinary support. PMID- 17426476 TI - Late-onset sarcoidosis after liver transplantation for primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) and systemic sarcoidosis are granulomatous diseases of unknown etiology whose hepatic manifestations may infrequently be imitative of one another. Described herein is the first reported case in the medical literature of systemic sarcoidosis developing after liver transplantation for PBC. The presented patient, who suffered from typical clinical, laboratory, and pathologic manifestations of PBC, developed decompensated liver cirrhosis within a course of 8 years, necessitating orthotopic liver transplantation. A year and a half after transplantation, the patient developed diffuse, biopsy proven, dermatologic and pulmonary manifestations of systemic sarcoidosis, which promptly responded to corticosteroid treatment. In retrospect, the patient's longstanding liver disease was probably caused by an unrecognizable, isolated hepatic form of sarcoidosis or an overlap between PBC and sarcoidosis. This patient illustrates the complexity that may be rarely encountered in differentiating between PBC and hepatic sarcoidosis. Discussed are the clinical, laboratory, and pathologic overlaps between hepatic sarcoidosis and PBC, and clues that may aid in the diagnosis and differentiation between the 2 disorders. Hepatologists and liver transplantation specialists should be aware of the rare possibility of hepatic sarcoidosis imitating PBC, and exacerbating systemically after liver transplantation. PMID- 17426477 TI - Septicemia, hepatic abscess, and encephalitis due to Yersinia enterocolitica. PMID- 17426478 TI - Primary biliary cirrhosis presenting with isolated eosinophilia. PMID- 17426479 TI - First reported case of a Candida glabrata perihepatic abscess as a complication of percutaneous endoscopic gastrostomy tube placement. PMID- 17426480 TI - Probiotics in the prevention of traveler's diarrhea: meta-analysis. PMID- 17426481 TI - Morphine-induced antinociception in the formalin test: sensitization and interactions with D1 and D2 dopamine receptors and nitric oxide agents. AB - In this study, the effects of dopamine receptor antagonists and nitric oxide agents on morphine-induced sensitization in the formalin test in mice have been investigated. Repeated daily intraperitoneal administration of morphine (30 mg/kg for 3 days) followed by a 11-day wash out period increased morphine-induced antinociception in the formalin test, which may be due to sensitization. The antinociceptive response to higher doses of morphine (6 and 9 mg/kg) but not 3 mg/kg was significantly increased in sensitized animals compared with control groups. Pretreatment of animals with an opioid receptor antagonist, naloxone (4 mg/kg), during repeated administration of morphine, attenuated the morphine induced sensitization. In the second part of the study, the animals received SCH23390 (D1 receptor antagonist), sulpiride (D2 receptor antagonist), L-Arg (nitric oxide precursor) and NG-nitro-L-Arg methylester (nitric oxide synthase inhibitor) during repeated morphine administration, to evaluate the role of dopamine receptor antagonists and nitric oxide agents in this phenomenon. Pretreatment of animals with NG-nitro-L-Arg methylester (20 mg/kg) and sulpiride (100 mg/kg) during morphine sensitization decreased the antinociceptive response to higher doses of morphine in the formalin test. It is concluded that D2 dopamine receptor and nitric oxide mechanisms may be involved at least partly in morphine-induced sensitization in the formalin test. PMID- 17426482 TI - Potentiation of olanzapine substitution in rats discriminating clozapine by the D2/3 agonist quinpirole. AB - The D2/3 agonist (+)-4-propyl-9-hydroxynaphthoxazine (PHNO) has been reported to enhance the ability of olanzapine to substitute for clozapine and attenuate olanzapine-induced response suppression in monkeys. These data suggest that the relatively marked D2/3 antagonist actions of olanzapine limit its substitution for clozapine. The work reported here replicated and extended these findings. Twelve rats were trained to discriminate clozapine (5 mg/kg, intraperitoneal) from vehicle in an FR30 quantal food rewarded assay. The substitution curve for olanzapine (0-2.5 mg/kg) was then computed after treatment with either vehicle or a high dose (0.1 mg/kg) of the D2/3 agonist quinpirole. The olanzapine substitution curve was shifted significantly 5.2-fold in parallel to the left by quinpirole. Olanzapine suppressed responding significantly, but this effect was not attenuated or enhanced by quinpirole, which suppressed responding itself. Thus antagonist actions at D2/3 receptors clearly limit the ability of olanzapine to substitute for clozapine. These findings suggest that the clozapine versus vehicle discrimination is probably a bioassay for agents that resemble clozapine but which do not necessarily induce D2/3 antagonism. This discrimination may therefore not specifically detect clozapine-like antipsychotics, although it may be of value in developing such antipsychotics. The low discriminability of antipsychotics in general may be because antagonist actions at D2/3 receptors limit incentive salience in discrimination assays. These data are compatible with recent theorizing that therapeutic actions of antipsychotics in schizophrenia involve D2/3 receptor-mediated attenuation of stimulus salience. PMID- 17426483 TI - Comparative effects of nonselective and subtype-selective gamma-aminobutyric acidA receptor positive modulators in the rat-conditioned emotional response test. AB - Benzodiazepine receptor anxiolytics show no selectivity between gamma aminobutyric acid-A receptors containing alpha1, alpha2, alpha3 or alpha5 subunits. Pharmacological studies and data emerging from transgenic mouse models, however, predict that compounds with selective affinity and/or efficacy for gamma aminobutyric acid-A receptor subtypes would have novel pharmacological profiles. Thus, the gamma-aminobutyric acid-A-alpha1 'affinity selective' drug zolpidem has a sedative-hypnotic profile, whereas L838,417, which has 'selective efficacy' for gamma-aminobutyric acid-A-alpha2, alpha3 and alpha5 receptors, has an anxiolytic like profile. Here, we compare the nonselective benzodiazepine-site-positive modulators diazepam, lorazepam, midazolam, alprazolam and zopiclone with (i) gamma-aminobutyric acid-AA-alpha1 affinity selective compounds zolpidem and CL218,872 and (ii) L838,417, in the rat-conditioned emotional response test after systemic administration. Given the role of the basolateral amygdala in anxiety and the expression of alpha1, alpha2 and alpha3 subunits in this region, we also assessed the effects of bilateral infusion of L838,417 and midazolam directly into basolateral amygdala in the conditioned emotional response test. Nonselective modulators at low-moderate doses produced anxiolytic effects and sedation at higher doses. Zolpidem was inactive as an anxiolytic and engendered severe sedation, whereas CL218,872 produced an anxiolytic-like profile with minimal sedation. L838,417 produced an anxiolytic-like profile with no sedation, albeit producing behavioural disturbance at high doses. Infusion of midazolam and L838,417 into basolateral amygdala engendered anxiolytic-like effects, although both compounds were more effective after systemic injections, implicating additional brain sites in their anxiolytic-like actions after systemic administration. In conclusion, the diversity of effects of the compounds studied implicates both intrinsic efficacy and/or subtype selectivity as important determinants of anxiolytic-like effects in the rat-conditioned emotional response test. PMID- 17426484 TI - Sensitization and cross-sensitization after chronic treatment with methylphenidate in adolescent Wistar rats. AB - An increasing debate exists about the potential of early exposure to methylphenidate to increase the risk for drug abuse. In addition, little is known about the neurobiological effects of early exposure to methylphenidate. This study was designed to investigate whether chronic treatment with methylphenidate induces behavioral sensitization to subsequent methylphenidate and D-amphetamine challenge in adolescent Wistar rats. Young Wistar rats (P25) were treated with either methylphenidate (1, 2, or 10 mg/kg, intraperitoneally) or saline for 28 days. After 14 days of washout, animals were challenged with methylphenidate 2.5 mg/kg intraperitoneally or D-amphetamine 2 mg/kg intraperitoneally (P67). Locomotor behavior was assessed using the open field test. Rats chronically treated with methylphenidate in the adolescent period showed augmented locomotor sensitization to D-amphetamine but not to methylphenidate in the adult phase. These findings suggest that early exposure do methylphenidate might increase the risk for subsequent D-amphetamine abuse. Further studies focusing on the neurobiological effects of early exposure to methylphenidate are warranted. PMID- 17426485 TI - Histaminergic H1 receptors mediate L-histidine-induced anxiety in elevated plus maze test in mice. AB - The central histaminergic system is reported to mediate behavioural, hormonal and physiological homeostasis of living organisms. Recent reports indicate its prominent role in various neurobehavioural disorders such as depression and psychosis. This study evaluated the effect of activation of the central histaminergic system in anxiety-like conditions, using the elevated plus-maze test in mice, and elucidated the role of different histaminergic receptors mediating such effects. Peripheral administration of L-histidine (L-His), in a dose-dependent manner, significantly decreased the exploration time in open arms and number of entries into open arms without modifying the number of entries into closed arms of the elevated plus-maze, indicating anxiogenesis. Further, such effects of central histamine were significantly attenuated, in a dose-dependent manner, by pretreatment with pyrilamine (H1 receptor antagonist). Pretreatment with either zolantidine (H2 receptor antagonist) or thioperamide (H3 receptor antagonist), however, failed to attenuate the L-His-induced anxiogenesis. Our results indicate that anxiogenic effects of central histaminergic system appear to be mediated prominently by activation of H1 receptors. PMID- 17426486 TI - Inhaled toluene vapor as a discriminative stimulus. AB - Few studies exist exploring the discriminative stimulus effects of inhalants and none that have trained an interoceptive discrimination using the inhaled route. This study was designed to assess if it was possible to train an inhaled toluene discrimination. The second objective was to determine whether the discrimination was based on interoceptive or exteroceptive stimulus effects. Eight B6SJLF1/J mice were trained to discriminate 10 min of exposure to 6000 ppm inhaled toluene vapor from air, using a standard food-reinforced operant procedure. Toluene vapor produced robust, concentration-dependent, discriminative stimulus effects, with concentrations of 4000 ppm and higher producing full substitution. Substitution of inhaled toluene vapor for the training condition was exposure-time dependent. A minimum of 7 min of exposure to 6000 ppm was required to produce complete substitution. Injected intraperitoneal toluene produced dose-dependent full substitution for inhaled toluene vapor. Both inhaled and intraperitoneal ethylbenzene produced similar levels of partial substitution for 6000 ppm toluene vapor. Inhaled isoflurane vapor produced no substitution for toluene vapor. These results show that a toluene vapor discrimination can be successfully trained in mice and the discrimination is selective for toluene compared to ethylbenzene and isoflurane. The results also suggest that the discrimination was likely to have been based primarily on interoceptive rather than exteroceptive stimulus effects. PMID- 17426487 TI - Protection against Abeta-induced memory loss by tripeptide D-Arg-L-Glu-L-Arg. AB - The molecular and cellular mechanisms underlying the memory deficits in Alzheimer's disease are increasingly thought to be associated with faulty processing of amyloid precursor protein. Following our earlier findings that it is possible to use the tripeptide RER (NH2-D-Arg-L-Glu-L-Arg-COOH, derived from the external domain of amyloid precursor protein) to rescue memory in animal models, we report here that the diasteromeric (D/L) form of the acetylated tripeptide RER protects against Abeta-induced memory loss for a passive avoidance task in young chicks and enhances retention for a weak version of the task when injected peripherally up to 12 h before training. The tripeptide readily crosses the blood-brain barrier, binds to membrane receptor sites in the brain and is without adverse effects on general behaviour. We discuss this finding in the context of other studies of the importance of peptides containing D-amino acids, and conclude that these RER-related peptides may form the basis for a potential therapeutic agent in the early stages of Alzheimer's disease. PMID- 17426488 TI - Oxidative stress in methamphetamine-induced self-injurious behavior in mice. AB - Previous studies have shown that N-methyl-D-aspartate, the formation of free radicals and poly(ADP-ribose) polymerase are related to methamphetamine-induced neurotoxicity. This study was designed to investigate the involvement of oxidative stress in methamphetamine-induced self-injurious behavior in mice. In this study, methamphetamine (20 mg/kg) induced continuous self-injurious behavior in six of seven mice. N-methyl-D-aspartate-receptor antagonists (MK801 and 3-((R) 2-carboxypiperazin-4-yl) propyl-1-phosphonic acid) significantly attenuated this methamphetamine-induced self-injurious behavior. These results suggest that the activation of N-methyl-D-aspartate receptors is involved in methamphetamine induced self-injurious behavior. Furthermore, we found that the nonselective nitric oxide synthase inhibitor l-N-nitro-L-arginine methyl ester hydrochloride and the neuronal nitric oxide synthase inhibitor 7-nitroindazole, but not the inducible nitric oxide synthase inhibitor aminoguanidine, the free-radical inhibitors fullerene and 3-methyl-1-phenyl-2-pyrazolin-5-one-186, or the poly(ADP ribose) polymerase inhibitor benzamide, significantly attenuated methamphetamine induced self-injurious behavior. The present results show that oxidative stress, which is mediated by the activation of neuronal nitric oxide synthase, is associated with methamphetamine-induced self-injurious behavior. These findings may help us to better understand the clinical phenomenon of self-injurious behavior. PMID- 17426489 TI - One-way avoidance learning and diazepam in female roman high-avoidance and low avoidance rats. AB - The goal of this experiment was to study the influence of the time spent in the safe compartment (30 vs. 1 s) and of an intraperitoneal injection of diazepam (1 mg/kg vs. vehicle) on one-way avoidance learning, in inbred female roman high avoidance and roman low-avoidance rats. Rats learned to run from a danger compartment, where they received a warning signal (88 dB tone) followed by a 1 mA electric footshock, to a safe compartment, where these stimuli were not presented. The number of trials needed to reach 10 consecutive avoidance responses was the dependent variable. Roman low-avoidance rats exposed to 1 s in the safe compartment showed poorer performance than their roman high-avoidance counterparts. These differences were not observed in rats exposed to 30 s in the safe place, and were abolished by the injection of diazepam. These results suggest the importance of fear and reinforcement in one-way avoidance learning and its usefulness for studying emotional processes underlying genetic or pharmacological manipulations. PMID- 17426490 TI - An initiative that sets the stage for the future: the editor's viewpoint. PMID- 17426492 TI - The home healthcare nurse of the 21st century: you've come a long way, baby! PMID- 17426493 TI - Looking back. A tribute to my great aunt, Marguerite Wales: author, leader, consultant, public health nurse, and director of Henry Street Visiting Nurse Service. PMID- 17426494 TI - Moving toward the future: providing speech-language pathology services via telehealth. PMID- 17426495 TI - Home care today: showcasing interdisciplinary management in home care. PMID- 17426496 TI - No place like home: a history of nursing and home care in the U.S. PMID- 17426497 TI - The pharmacologic management of pain. A home care requisite. PMID- 17426499 TI - Health coaching: a new and exciting technique to enhance patient self-management and improve outcomes. AB - Health coaching is quickly emerging as a new approach of partnering with patients to enhance self-management strategies for the purpose of preventing exacerbations of chronic illness and supporting lifestyle change. Medicare is now pilot testing this approach for patients with congestive heart failure and diabetes. With acute care hospitalization an outcome of great interest to us all, health coaching is an exciting technique worthy of consideration by home health providers. PMID- 17426501 TI - Highlights of the times. Ten roller coaster years in home care: 1995 to 2005. PMID- 17426502 TI - Memories are made of this! PMID- 17426504 TI - Development of pain: maturation of spinal inhibitory networks. PMID- 17426505 TI - Genetics of pain: lessons for future studies. PMID- 17426506 TI - Cytokines, inflammation, and pain. PMID- 17426507 TI - Imaging of pain. PMID- 17426508 TI - Viral vector-based gene transfer for treatment of chronic pain. PMID- 17426509 TI - Signal transduction in chronic pain. PMID- 17426510 TI - Spinal mechanisms of pain. PMID- 17426511 TI - Impaired insulin signaling as a potential trigger of pain in diabetes and prediabetes. PMID- 17426512 TI - Ion channels in pain transmission. PMID- 17426513 TI - Animal models of pain. PMID- 17426515 TI - Use of the Beryllium Lymphocyte Proliferation Test (BeLPT) for screening. PMID- 17426518 TI - Malignant mesothelioma in telephone workers. PMID- 17426519 TI - High job strain and ambulatory blood pressure in middle-aged men and women from the Belgian job stress study. AB - OBJECTIVE: The aim of this study was to assess whether job strain is associated with 24-hour ambulatory blood pressure measurements within a subsample of the Belgian Job Stress Project (BELSTRESS) population. METHODS: A group of 89 middle aged male and female workers perceiving high job strain and an equally large group of workers perceiving no high job strain wore an ambulatory blood pressure monitor for 24 hours on a regular working day. RESULTS: Mean ambulatory blood pressure at work, at home, and while asleep was significantly higher in workers with job strain as compared with others. The associations between job strain and ambulatory blood pressure were independent from the covariates. CONCLUSIONS: Within this study, high job strain was an important independent risk factor for higher ambulatory blood pressure at work, at home, and during sleep in a group of men and women. PMID- 17426520 TI - Indirect cost burden of migraine in the United States. AB - OBJECTIVE: The purpose of this study was to determine the indirect cost burden associated with migraine. METHODS: Data were obtained from Thomson-Medstat's Health and Productivity Management (HPM) database for the 2002 through 2003 calendar years. The migraine cohort was composed of patients who had a diagnosis of migraine or migraine-specific abortive prescription medication, or both. A control cohort of patients without migraine was matched to patients in the migraine cohort. The average annual indirect burden of illness (BOI) of migraine and a national indirect BOI were estimated. RESULTS: Annual indirect expenditures were significantly higher in the migraine group compared with the control group ($4453 vs $1619; P<0.001). The national annual indirect BOI, excluding presenteeism, was estimated to be $12 billion (mostly attributed to absenteeism). CONCLUSIONS: Migraine imparts a substantial indirect cost burden. Projected to a national level, this amounts to an annual cost to US employers of approximately $12 billion. PMID- 17426521 TI - Cyclic load magnitude is a risk factor for a cumulative lower back disorder. AB - OBJECTIVE: Epidemiological data suggest that high loads lifted by workers engaged in static and cyclic daily activities may be a risk factor for low back disorder. Our previous research provided physiological and biomechanical validation of the epidemiological data for static load conditions. The objective of this report was to provide physiological and biomechanical experimental validation to the epidemiological data in cyclic (repetitive) load conditions. METHODS: Three groups of in vivo feline models were subjected to 3 cyclic load levels in a series of 6 periods of 10 minutes of work spaced by 10 minutes of rest followed by 7 hours of rest. Multifidus electromyography (EMG) and lumbar displacement were statistically analyzed after processing. RESULTS: Delayed muscular hyperexcitability was observed only in moderate (40 N) and high (60 N) loads (P<0.0001) but was absent in low (20 N) loads. The magnitude of the delayed hyperexcitability was found to be higher (P<0.0001) in the high (60 N) loads compared with the moderate (40 N) loads. CONCLUSIONS: Exposure to moderate and high loads in cyclic (repetitive) work results in an acute neuromuscular disorder indicative of soft tissue inflammation that may become chronic with further exposure. PMID- 17426522 TI - Occupational medicine physicians in the United States: demographics and core competencies. AB - OBJECTIVE: We assessed the demographic profile and opinions of current occupational medicine (OM) physicians on the importance of specific core competencies. METHODS: A random sample of 1500 OM physicians listed in the membership directory of the American College of Occupational and Environmental Medicine (ACOEM) were asked to complete a voluntary survey. RESULTS: Six hundred and ten OM physicians completed the survey. Fifty two percent worked in clinical settings, and 16% worked in corporate or industrial settings. Eighty percent were satisfied with their choice of careers. CONCLUSIONS: OM physicians appeared to be highly trained, with 60% certified in OM and 68% board certified in other specialties. The OM physicians valued staying current in the field, understanding the relationship between occupational exposure and health, and communicating with stakeholders most highly. Occupational physicians are an important source of knowledge regarding what competencies and core knowledge areas are important for OM practice. PMID- 17426523 TI - Occupational injury surveillance of traumatic injuries in Illinois, using the Illinois trauma registry: 1995-2003. AB - BACKGROUND: Trauma registries continue to be underutilized for surveillance, despite providing data on the most severe injuries with a level of detail not available in national data sets or workers' compensation files. METHODS: We evaluate trends and patterns of traumatic occupational injuries from the Illinois Trauma Registry (ITR). RESULTS: Between 1995 and 2003, 44.4 of every 100,000 Illinois workers (age-adjusted) suffered work-related, nonfatal traumatic injuries. The majority of workers suffering traumatic injuries were white males younger than 55 years old. Falls were the most common cause of injury, and fracture of the extremities was the most common type of injury experienced by Illinois workers. Approximately 8% of all workers required either partial or total ambulatory assistance at time of discharge. CONCLUSION: The ITR provides detailed and complete data regarding the most severe occupational injuries. CLINICAL SIGNIFICANCE: In contrast to reports from national surveillance data sets, we do not observe a significant decline in occupational injuries between 1995 and 2003. Trauma registries should be used more frequently for surveillance programs, because they provide detailed and reliable data regarding the most severe occupational injuries not available in other data sets. PMID- 17426524 TI - Implications of detecting the mold Syncephalastrum in clinical specimens of New Orleans residents after Hurricanes Katrina and Rita. AB - After the extensive flooding in New Orleans following Hurricanes Katrina and Rita, thousands of homes in the flooded areas had significant growth of mold. The potential health effects from exposures to these extraordinary environments are unknown. In February 2006, we investigated a cluster of patients with clinical specimens yielding Syncephalastrum, a zygomycete that rarely causes infection. We identified the cases of eight patients from September 12, 2005, to January 12, 2006, with specimens from sputum, bronchoalveolar lavage, endotracheal aspirate, ear swab, and nasal swab. All patients appeared to be transiently colonized without evidence of infection, even among immunosuppressed patients. Only one patient reported significant exposure to mold (working on mold remediation without wearing a respirator) on the day of his incident culture. PMID- 17426525 TI - Employee working conditions and healthcare system performance: the Veterans Health Administration experience. AB - OBJECTIVE: The authors explored the association between health care employees' perceptions of their organizations and objective measures of system performance. METHODS: A national survey of employees conducted in 2001 by the Veterans Administration (VA) assessed employee perceptions of hospital organizational characteristics. The authors analyzed cross-sectional associations between these perceptions and objective measures of health care system performance--employee and patient care outcomes. RESULTS: Employee perceptions of organizational climate (indicators of the organizational culture) were strongly related to overall satisfaction and measures of system performance. Overall, change in perceptions of organizational climate by 1 standard deviation (SD) was potentially associated with changes of 2% to 35% in selected outcomes. CONCLUSIONS: Organizational climate, policies, and resultant working conditions in health care institutions appear to be strong drivers of system performance. Interventions directed toward improving care quality and safety should address these factors. PMID- 17426526 TI - Dermal absorption of solvents as a major source of exposure among shipyard spray painters. AB - OBJECTIVE: The purpose of this study was to evaluate the relevance of inhalational and dermal exposure to solvents in shipyard spray painters. Special emphasis was placed on the spatial distribution of dermal exposure and absorption across different regions of the body. METHODS: Fifteen male spray painters were recruited for this study. The subjects were monitored during a 3-day work period using a repeated-measures study design. Air and dermal exposure of solvents were collected each day. Urine was collected before and after the work shift. RESULTS: Air samples showed that the workers were primarily exposed to ethylbenzene and xylene. The concentrations of ethylbenzene and xylene outside the workers' masks were 59.2 +/- 10.4 (mean +/- standard error [SE]) ppm and 29.4 +/- 4.70 ppm, whereas those inside the masks were 7.91 +/- 17.4 ppm and 3.83 +/- 8.22 ppm, respectively. The average mass of ethylbenzene and xylene across the different body regions inside the block units of assembled ships were 305.1 +/- 63.9 mg and 165.6 +/- 34.1 mg. The quantity was, on average, 5.8 and 5.1 times higher than those collected outside the blocks. In both measurements, the highest exposure mass was found on the upper legs, and the lowest exposure mass was found on the back. Principal component analysis (PCA) was used to transform the variables of dermal exposure for all investigated body regions into only one principal component. Multiple regression analyses revealed a significant relationship between dermal exposure to xylene (PCA dermal xyl) and urinary methylhippuric acid (MHA) levels, adjusting for air xylene exposure (R2=0.491, P<0.05). CONCLUSIONS: :The present study indicated that dermal exposure to xylene significantly increased the urinary levels of MHA, suggesting that dermal exposure to solvents was an important route among spray painters. PMID- 17426527 TI - Tobacco smoking by occupation in Australia: results from the 2004 to 2005 National Health Survey. AB - OBJECTIVE: This study presents the most recent estimates of Australia's national tobacco smoking rates by occupation. METHODS: Smoking data was extracted from the 2004 to 2005 National Health Survey, which captured approximately 26,000 persons and achieved a response rate of around 90%. Participants were limited to those of working age (18 to 64 years), with data stratified by job category and gender during the analysis. RESULTS: The prevalence of smoking among Australian workers is estimated to be 25% (28% among males and 21% among females). Tobacco usage is considerably less common among those who are employed compared with the unemployed. By job category, smoking was most common among laborers and the least common among professionals, managers, or administrators. CONCLUSIONS: Overall, this study suggests that Australian rates of tobacco smoking vary widely depending on occupation. Effective tobacco-control strategies targeting vulnerable sections of the workforce, particularly blue-collar workers, are clearly needed. PMID- 17426528 TI - TNF-alpha polymorphisms in chronic beryllium disease and beryllium sensitization. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) is a potent cytokine involved in normal immune functions. The aim of this study was to investigate if there is an association between chronic beryllium disease or beryllium sensitization and two variants of the TNF-alpha gene located at -308 and -238 called TNF-alpha 308*02 and TNF-alpha-238*02. METHODS: TNF-alpha-308 and TNF-alpha-238 genotyping was conducted in a large, population-based cohort consisting of 886 beryllium workers (92 individuals with chronic beryllium disease, 64 who were beryllium sensitized, and 730 individuals without sensitization or disease). RESULTS: The odds of chronic beryllium disease in the presence of at least one TNF-alpha 308*02 or TNF-alpha-238*02 allele was not significant (OR=1.0; 95% CI=0.7, 1.7 and OR=0.8; 95% CI=0.4, 1.6). This was true regardless of whether a worker was homozygous or heterozygous for TNF-alpha-308*02 or TNF-alpha-238*02. Similarly, neither allele was associated with sensitization (P>0.05). CONCLUSIONS: Unlike an earlier report, there was no association between these specific TNF-alpha alleles and either chronic beryllium disease or sensitization to beryllium. PMID- 17426529 TI - An employer-based cost-benefit analysis of a novel pharmacotherapy agent for smoking cessation. AB - INTRODUCTION: An employer-based cost-benefit analysis for varenicline versus bupropion was conducted using clinical outcomes from a recently published trial. METHODS: A decision tree model was developed based on the net benefit of treatment to produce a nonsmoker at 1 year. Sensitivity analyses were conducted based on quit rates with placebo and varenicline and the cost of varenicline. RESULTS: Estimated 12-month employer cost savings per non-smoking employee were $540.60 for varenicline, $269.80 for bupropion SR generic, $150.80 for bupropion SR brand, and $81.80 for placebo. Varenicline was more cost beneficial than placebo, which had quit rates of 16.9% or less. The quit rate with varenicline would have to be 100 bpm, has been utilized as a physical sign of hypovolemic shock among the injured for decades without evidence to support its use as a predictor of injury or significant hypovolemia. We sought to determine whether admission heart rate is a valid predictor of hemodynamically significant injuries. METHODS: Trauma registry data from 1998 to 2004 were analyzed with logistic regression to determine whether heart rate was associated with need for emergent intervention for bleeding (laparotomy, thoracotomy, or angiography), need for packed red blood cell (pRBC) transfusion in the first 24 hours, or severe injury (ISS >25) after blunt or penetrating trauma. RESULTS: Records of 10,825 patients were analyzed. Overall, heart rate was neither sensitive nor specific in determining the need for emergent intervention, pRBCs in the first 24 hours or severe injury. This was not altered by the presence of hypotension (systolic blood pressure <90 mm Hg) or age in the blunt cohort. CONCLUSIONS: Heart rate alone is not sufficient to determine the need for emergent interventions for hemorrhage. Although tachycardia may still indicate need for emergent intervention in the trauma patient, its absence should not allay such concern. PMID- 17426535 TI - Hypertonic saline and pentoxifylline attenuates gut injury after hemorrhagic shock: the kinder, gentler resuscitation. AB - BACKGROUND: We have previously demonstrated that postshock resuscitation with Hypertonic saline and Pentoxifylline (HSPTX) attenuates pulmonary and histologic gut injury when compared with Ringer's lactate (RL). In this study, we hypothesized that the decrease in gut injury observed with HSPTX is associated with the attenuation of inducible nitric oxide synthase (iNOS) activity and production of ileal proinflammatory mediators after hemorrhagic shock. METHODS: In a rat model of hemorrhagic shock, resuscitation was conducted with RL (32 mL/kg; n = 7) or HSPTX (4 mL/kg 7.5% NaCl + PTX 25 mg/kg; n = 7). Sham animals that did not undergo shock were also studied. Four hours after resuscitation, the terminal ileum was collected for evaluation of nitrite, tumor necrosis factor (TNF)-alpha, Interleukin (IL)-6, and cytokine-induced neutrophil chemoattractant (CINC) by enzyme immunoassay. Heme oxygenase-1 (HO-1), iNOS, cytoplasmic inhibitor of kappa B (Ikappa B) phosphorylation, and nuclear factor (NF)kappa B p65 nuclear translocation were determined by Western blot. RESULTS: HSPTX resuscitation resulted in a 49% decrease in iNOS when compared with RL (p < 0.05). Similar results were obtained when examining nitrite (882 +/- 59 vs. 1,435 +/- 177 micromol/L; p < 0.01), and HO-1 content (p < 0.05). RL resuscitation resulted in markedly higher levels of TNF-alpha (83 +/- 27 vs. 9 +/- 5 pg/mL; p < 0.01), IL-6 (329 +/- 58 vs. 118 +/- 43 pg/mL; p < 0.05), and CINC (0.43 +/- .06 vs. 0.19 +/- .08 ng/mL; p < 0.05) than HSPTX. The increase in cytokines observed with RL was also associated with an increase in I-kappaB phosphorylation (p < 0.01) and NF-kappaB p65 nuclear translocation (p < 0.001). CONCLUSION: The attenuation in gut injury after postshock resuscitation with HSPTX is associated with downregulation of iNOS activity and subsequent proinflammatory mediator synthesis. HSPTX has the potential to be a superior resuscitation fluid with significant immunomodulatory properties. PMID- 17426536 TI - A ten-year retrospective review: does pyloric exclusion improve clinical outcome after penetrating duodenal and combined pancreaticoduodenal injuries? AB - OBJECTIVES: We sought to determine whether the performance of pyloric exclusion during repair of penetrating advanced duodenal injuries prevents postoperative duodenal fistulas and improves clinical outcome. METHODS: A retrospective chart review of patients from 1995 to 2004 with penetrating duodenal injuries >or=grade II and all combined pancreaticoduodenal injuries was performed. Patients managed either without or with pyloric exclusion were compared on the basis of age, sex, mechanism, injury grade, Injury Severity Score (ISS), hemodynamic stability, the presence of vascular injury or associated injuries, postoperative complications, length of hospital stay, and mortality. RESULTS: Fifteen of 29 patients were managed without pyloric exclusion and 14 with exclusion. Both groups were similar with respect to age, sex, mechanism, injury grade, ISS, hemodynamic stability, the presence of vascular injury, associated abdominal injuries, and mortality rates. A trend toward a higher overall complication rate (71% vs. 33%), pancreatic fistula rate (40% vs. 0%), and length of hospital stay (24.3 days vs. 13.5 days) was evident in the pyloric exclusion group. No duodenal fistula was detected in either patient group. CONCLUSION: In our study population, the performance of pyloric exclusion for penetrating advanced duodenal injury and combined pancreatic and duodenal injuries did not improve clinical outcome. The trend toward a greater overall complication rate, pancreatic fistula rate, and increased length of hospital stay in the pyloric exclusion group suggests that simple repair without pyloric exclusion is both adequate and safe for most penetrating duodenal injuries. PMID- 17426537 TI - Preperitonal pelvic packing for hemodynamically unstable pelvic fractures: a paradigm shift. AB - BACKGROUND: The current management of pelvic fracture patients who are hemodynamically unstable in the United States consists of aggressive resuscitation, mechanical stabilization, and angioembolization. Despite this multidisciplinary approach, our recent analysis confirms an alarming 40% mortality in these high-risk patients. Therefore, we pursued alternate therapies to improve patient outcomes. European trauma groups have suggested the technique of pelvic packing via laparotomy to directly address the venous bleeding that comprises 85% of pelvic fracture hemorrhage. We hypothesized that a modified technique of direct preperitoneal pelvic packing (PPP) would reduce the need for angiography, decrease blood transfusion requirements, and lower mortality. METHODS: Since September 2004, all patients at our ACS-verified level I trauma center with hemodynamic instability and pelvic fractures underwent PPP/external fixation, according to our protocol. Statistics are reported as mean +/- SEM and analyzed using Student's t test. RESULTS: During the study period, 28 consecutive patients underwent PPP. There was one protocol deviation of prePPP angiography to evaluate an extremity vascular injury. The majority were men (68%) with a mean age of 40 +/- 3.9 years and a mean injury severity score of 55 +/- 3.0. The mean emergency department (ED) systolic blood pressure was 77 +/- 3.0 mm Hg, heart rate was 120 +/- 4.3 bpm, and base deficit 13 +/- 0.8 mmol/L. Pelvic fracture classifications included lateral compression (LC) II (9), anteroposterior compression (APC) III (8), LC I (3), vertical shear (3), LC III (3), and APC II (2). Patients required 4 +/- 1.2 units of packed red blood cells (PRBCs) during 82 +/- 13 minutes in the ED. Blood transfusion requirements before postoperative surgical intensive care unit (SICU) admission compared with the subsequent 24 postoperative hours were significantly different (12 +/- 2.0 versus 6 +/- 1.1; p = 0.006). The first 4 patients underwent routine angiography postPPP, with 1 undergoing therapeutic embolization; 4 of the subsequent 24 patients underwent angioembolization with clinical concern of ongoing pelvic hemorrhage. Seven (25%) patients died from multiple organ failure (2), postinjury myocardial infarction/pulseless electrical activity (PEA) arrest (2), invasive mucormycosis (1), withdrawal of care (1), and closed head injury (1); there were no deaths as a result of acute blood loss. CONCLUSIONS: PPP is a rapid method for controlling pelvic fracture-related hemorrhage that can supplant the need for emergent angiography. There is a significant reduction in blood product transfusion after PPP, and this approach appears to reduce mortality in this select high-risk group of patients. PMID- 17426538 TI - Extraperitoneal pelvic packing: a salvage procedure to control massive traumatic pelvic hemorrhage. AB - OBJECTIVE: To describe the method of extraperitoneal pelvic packing (EPP), and to assess the impact of EPP on outcome in severely hemodynamically unstable patients after blunt pelvic trauma. METHODS: Of 661 patients treated for pelvic trauma, 18 underwent EPP as part of our protocol with the intent to control massive pelvic bleeding and constituted the study population. Data retrospectively collected from the medical records and from the Ulleval Trauma Registry included demographics, fracture classification, additional injuries, blood transfusions, surgical interventions, angiographic procedure, physiologic parameters, and survival. RESULTS: Survival rate within 30 days was 72% (13/18), and correlated inversely to the age of the patient (p = 0.038). Only one of the nonsurvivors died of exsanguination. A significant increase in systolic blood pressure (BP) (p = 0.002) was observed immediately after EPP. Angiography performed after EPP was positive for arterial injury in 80% of patients. All types of pelvic ring fractures were represented. CONCLUSIONS: EPP as part of a multi-interventional resuscitation protocol might be life saving in patients with life-threatening pelvic injury who are exsanguinating. However, the high rate of arterial injuries seen after EPP indicates that the procedure should be supplemented with angiography once the patient is sufficiently stabilized to tolerate transportation to the angiography suite. PMID- 17426539 TI - Diagnostic peritoneal lavage: is 100,000 RBCs a valid figure for penetrating abdominal trauma? AB - BACKGROUND: Controversy exists regarding the interpretation of diagnostic peritoneal lavage results. This is especially true in the evaluation of patients sustaining penetrating trauma, specifically stab wounds to the lower chest and abdomen. Ideally one wants to avoid missed injuries and minimize unnecessary operations. METHODS: This is a retrospective review of 195 patients sustaining stab wounds to the anterior lower chest and abdomen at Parkland Memorial Hospital between 1993 and 2005, looking at missed injuries and false positive rates using red cell counts of 100,000, 10,000, and the standard criteria for blunt trauma including >500 white blood cells (WBCs), amylase, and/or bile. RESULTS: The first analysis used >100,000 red blood cells (RBCs)/mm3 as a positive value. The false positive rate was 12.2%. The second analysis used >10,000 RBCs/mm3 as a positive value with a false positive rate of 44%. When considering the entire study population (195 patients), the false positive rate increased when using the lower number (>10,000) from 2.5% to 15.8% (p < 0.001). There were no missed injuries when using >100,000 red cells and/or >500 white cells, the presence of bile or amylase. CONCLUSION: Decreasing the red blood cell count from >100,000 to >10,000 as the criteria for operating on patients with stab wounds to the anterior lower chest and/or abdomen will significantly increase the number of nontherapeutic procedures. Based on this study, >100,000 RBCs/mm3 appears to be a valid and safe number to use when evaluating these patients, particularly when used with other positive criteria such as increased white cells, bile, and amylase. PMID- 17426540 TI - Severity of burn injury and sepsis determines the cytokine responses of bone marrow progenitor-derived macrophages. AB - BACKGROUND: Although thermal injury and sepsis result in enhanced monocytopoiesis, the functional characteristics of macrophages that develop in the microenvironment of burn and sepsis are unknown. Here we compare cytokine responses of bone marrow progenitor-derived macrophages (BMO) and peritoneal macrophages (PMO) after graded levels of thermal injury and sepsis. METHODS: Mice were randomly divided into sham (S), burn (B), and burn sepsis (BS) groups. The mild injury group received either a 7-second dorsal scald burn alone or in combination with 1,000 colony forming units (CFU) Pseudomonas aeruginosa at the wound site. The severe injury group was subjected to a 10-second burn with or without inoculation of 5,000 CFU P. aeruginosa. ER-MP12+ progenitors were separated from bone marrow cells 72 hour after injury. Macrophage colony stimulating factor (M-CSF) and Granulocyte-macrophage colony stimulating factor (GM-CSF) responsive clonogenic potentials, and lipopolysaccharide (LPS) stimulated cytokine production were determined. RESULTS: In mild injury and sepsis, GM-CSF and M-CSF responsive clonal growth of ER-MP12+ progenitors was enhanced in the B and BS groups compared with the S group. M-CSF responsive colony growth in severe sepsis was significantly higher than that in all the other groups. LPS-stimulated tumor necrosis factor-alpha and Interleukin-6 levels were higher in the B and BS groups compared with the S group. Severe injury and sepsis attenuated this response significantly. The cytokine responses of PMO from both injury groups were similar to that of BMO. CONCLUSION: Severity of burn injury and the magnitude of sepsis influence the cytokine responses of BMO and PMO in a similar manner suggesting the microenvironment of burn injury and sepsis profoundly influence the functional phenotype of BMO. PMID- 17426541 TI - Recombinant activated factor VII increases survival time in a model of incompressible arterial hemorrhage in the anesthetized pig. AB - BACKGROUND: Hemorrhage is the leading cause of death in battlefield casualties and the second leading cause of death after civilian trauma. Evacuation time for military casualties to surgical care can be prolonged and improved hemostasis could greatly reduce mortality. There are several anecdotal reports that recombinant activated factor VII (rFVIIa) may arrest uncontrolled bleeding after trauma. However, the majority of prospective randomized controlled trials show little benefit in survival. The aim of this study was to determine whether rFVIIa could increase survival time within a clinically relevant time scale for military practice and reduce the volume of blood loss in a model of incompressible arterial hemorrhage. A secondary aim was to determine the effects of hypotensive versus normotensive resuscitation on the effectiveness of rFVIIa. METHODS: Terminally anaesthetized Large White pigs were randomly allocated to one of four treatment groups. All animals received a controlled hemorrhage of 40% of the total estimated blood volume. They were given either rFVIIa (180 microg/kg) or placebo (saline 0.3 mL/kg) intravenously and a 4 to 5 mm longitudinal aortotomy created in the infra renal aorta before resuscitation commenced with 0.9% saline to one of two target systolic arterial blood pressures (SBPs): 110 mm Hg (normotensive) or 80 mm Hg (hypotensive). Group sizes were as follows: placebo/normotensive (6), placebo/hypotensive (7), rFVIIa/normotensive (7), and rFVIIa/hypotensive (7). Survival was monitored for a maximum of 6 hours after the onset of resuscitation. RESULTS: rFVIIa was associated with a significantly prolonged survival time in animals managed hypotensively (214 [79-349] vs. 35 [19 52] minutes mean [95% confidence interval] rFVIIa vs. placebo, p = 0.03 Peto log rank test). There was no significant difference in survival time between those given rFVIIa and placebo in groups managed normotensively (128 [6-249] vs. 40 [15 66] minutes respectively, p = 0.27). Both rFVIIa and hypotensive management were associated with reduced uncontrolled hemorrhage volumes. There was no evidence of inappropriate intravascular thrombi or microthrombi associated with the use of rFVIIa. CONCLUSIONS: rFVIIa, combined with hypotensive resuscitation, can increase survival time and reduce hemorrhage in a model of arterial hemorrhage. The increase in survival time is clinically relevant for military evacuation of battlefield casualties to surgical care. PMID- 17426542 TI - Pretreatment with pro- and synbiotics reduces peritonitis-induced acute lung injury in rats. AB - BACKGROUND: To study whether enteral pretreatment with a synbiotic composition of lactic acid bacteria and bioactive fibers can reduce peritonitis-induced lung neutrophil infiltration and tissue injury in rats. MATERIALS AND METHODS: Rats were divided into five groups, and subjected to induction of peritonitis-induced lung injury using a cecal ligation and puncture model (CLP). All animals were pretreated for 3 weeks prior the CLP by daily gavage with either (1) a synbiotic composition (10(10) CFU of Pediococcus pentosaceus 5-33:3, 10(10) CFU of Leuconostoc mesenteroides 77:1, 10(10) CFU of L. paracasei subspecies paracasei, 10(10) CFU of L. plantarum 2362 plus fermentable fibers), (2) fermentable fibers alone, (3) nonfermentable fibers, (4) a probiotic composition (10(10) CFU of P. pentosaceus 5-33:3, 10(10) CFU of L. mesenteroides 77:1, 10(10) CFU of L. paracasei subsp. paracasei, 10(10) CFU of L. plantarum 2,362), or (5) a heat killed probiotic composition. All animals were killed 24 hours after CLP and lung tissue samples were studied for degree of neutrophil infiltration and levels of tumor necrosis factor (TNF)-alpha, Interleukin (IL)-1beta. In addition the lung wet-to-dry tissue weight ratio, the myeloperoxidase activity, and malondialdehyde content were also assessed. RESULTS: No mortality was encountered in any of the groups. Histologic signs of lung injury (number of neutrophils and TNF-alpha, IL 1beta staining) were observed in all groups except the synbiotic and probiotic treated groups. Myeloperoxidase activity and malondialdehyde content were significantly lower in the two lactobacillus- pretreated groups, with no difference between them. Heavy infiltration of lung tissue with neutrophils was observed only in fiber-treated (302.20 +/- 7.92) and placebo-treated (266.90 +/- 8.92) animals. This was totally abolished in the synbiotic-treated group (34.40 +/- 2.49). Lung edema (wet-to-dry lung weight ratio) was significantly reduced in the synbiotic-treated group (4.92 +/- 0.13 vs. 5.07 +/- 0.08 and 5.39 +/- 0.10, respectively). CONCLUSION: Three weeks of preoperative enteral administration of a synbiotic composition reduced peritonitis-induced acute lung injury in rats in a CLP model. PMID- 17426543 TI - Therapeutic endovascular treatments for traumatic vertebral artery injuries. AB - OBJECTIVE: Traumatic vertebral artery injuries pose difficulty in early diagnosis and management because of concomitant neurologic dysfunction and limitations in direct surgical access. The purpose of this report is to review endovascular therapy in patients with traumatic vertebral artery injuries for preservation of the parent artery, and to determine the safety and efficacy of such endovascular therapy. METHODS: Six patients with traumatic vertebral artery lesions were treated using therapeutic endovascular methods. Endovascular therapy was accomplished by stent insertion or coil embolization or both. RESULTS: Except one patient who underwent coil embolization of a transected vertebral artery, all dissections and pseudoaneurysms were successfully treated by stent placement or stent-assisted coiling with preservation of parent arteries. No additional surgical procedures for vascular lesions were required. There were no delayed neurologic or vascular complications and no lesions recurred during the follow-up period (mean, 36.7 months). CONCLUSION: The author's experience demonstrates that endovascular therapy using stents and coils is both feasible and safe in the treatment of traumatic vertebral artery injuries. Endovascular therapy selectively eliminated the vascular abnormality while maintaining the normal patency of the cerebral arteries. Long-term follow-up review of these repairs will be necessary to provide a full evaluation of the safety and efficacy of these devices. PMID- 17426544 TI - The utility of the anterolateral thigh donor site in reconstructing the United States trauma patient. AB - BACKGROUND: Identification of a single donor site capable of providing all the components of the soft tissue envelope and the ability to selectively harvest a subset of these components is a central requirement for the microvascular reconstruction of the trauma patient. The anterolateral thigh (ALT) flap's long pedicle and adaptability in supporting a variety of tissues (muscle, fascia, soft tissue) make it a valuable tool for microsurgical reconstruction in these challenging patients. We investigated the utility of the ALT as a donor for microvascular tissue reconstruction in a Level I trauma center. METHODS: We conducted a retrospective chart review on all trauma patients treated by the plastic surgery service at the R Adams Cowley Shock Trauma Center who required microsurgical free flap coverage from July 2002 to March 2005. Fifty-eight patients underwent reconstruction of traumatic deformities with 62 microvascular free flaps from the ALT region. RESULTS: Of the 58 patients, 42 were male and 16 were female with an average age of 39 years. Recipient site locations for the 62 flaps were lower extremity, upper extremity, trunk, and head and neck. Analysis of flap anatomy revealed that 43 were fasciocutaneous, 14 were myocutaneous, 2 were adipofascial, and 3 were myofascial (vastus lateralis muscle). Six flaps were based on septocutaneous perforators, whereas the remainder contained myocutaneous perforators. Nine thigh donor sites required a split thickness skin graft, and 53 were closed primarily. The size of the flaps ranged from 36 cm2 to 600 cm2. CONCLUSIONS: The ALT is a predictable donor site that facilitates a 2 team approach. ALT displays minimal donor site morbidity and in most cases provided sufficient tissue to cover the entire traumatic defect. Our results suggest the ALT is a reliable tissue source and an ideal donor site for the management of complex traumatic wounds in the United States. PMID- 17426545 TI - The value of multidetector row computed tomography in the diagnosis of traumatic clivus epidural hematoma in children: a three-year experience. AB - BACKGROUND: Clivus epidural hematoma (CEH) is reported to be extremely rare. However, we consider that CEH may not be as rare as commonly thought, and that it may often go unrecognized because of diagnostic problems. The aim of this study was to evaluate the value of multidetector row computed tomography (MDCT) in the diagnosis of CEH. METHODS: Twenty-seven children aged less than 16 years who had experienced traffic crashes or falls were retrospectively evaluated with MDCT including sagittal reconstruction of the cervical spine. RESULTS: Sagittal reconstructed images showed definitive findings of CEH in 3 (13%) of the 24 traffic-crash patients. CONCLUSION: MDCT can show definitive findings of CEH, and diagnostic problems can be solved through the utilization of MDCT. We consider that CEH may be more common than previously thought. PMID- 17426546 TI - Use of low-intensity pulsed ultrasound for posttraumatic nonunions of the tibia: a review of patients treated in the Netherlands. AB - BACKGROUND: Low-intensity pulsed ultrasound is effective in fresh fracture healing, resulting in a 40% reduction in healing time. The aim of this study is to determine the effect of ultrasound treatment on established tibia nonunions. METHODS: The study group consists of all Dutch patients of posttraumatic consecutive nonunion of the tibia, who started their ultrasound treatment between January 2000 and February 2003. In total, 71 cases have been included, which involve 56 men and 15 women. Mean age was 40 years. Low-intensity pulsed ultrasound was the only new treatment. Strict criteria of enrollment minimized any spontaneous healing chance. According to literature, the spontaneous healing rate was between 5% and 30%. The study outcome, healed or failed, was the primary efficacy parameter. Thirty percent was chosen to represent the maximum expected spontaneous healing and was the basis for statistical evaluation. Stratification was performed for the variables at the ultrasound treatment start. RESULTS: The overall healing rate is 52 of 71 cases (73%). Ultrasound treatment shows a statistical significant higher healing rate compared with that of the spontaneous healing chance (p < 0.0001). Stratification shows no statistical significance for any of the variables analyzed. The long-term follow-up shows high compliance rate and no refractures. CONCLUSION: Tibia nonunions have a high occurrence rate and cause significant impairment to daily functioning. This study shows that low intensity pulsed ultrasound is effective in the treatment of established tibia nonunions and can be seen as a good, safe, and cheaper alternative to surgery. PMID- 17426547 TI - Long bone fractures caused by penetrating injuries in terrorists attacks. AB - BACKGROUND: High-energy penetrating injuries are increasingly common in the civilian setting. During the years 2000 to 2003, more than 70 suicide bombing attacks occurred in Israel. These were characterized by high numbers of casualties, primarily caused by blast injuries. Injury caused by a blast can be either a primary blast effect of acceleration-deceleration of the blast wave or a secondary effect of metal fragments deliberately placed in explosives, causing severe penetrating injuries. The latter type of injury may result in severe open limb fractures. METHODS: We identified and reviewed 91 patients with 117 long bone fractures caused by penetrating terror-related injuries treated in our institution during 2000 to 2003. The patients were divided according to the mechanism of injury, i.e., either gunshot injury or blast injuries; several parameters were compared. RESULTS: Patients in the blast injury group included more children and elderly patients than the gunshot injury group did. This group also had a significantly higher Injury Severity Score and a higher number of associated injuries, including multiple fractures and mortality. The treatment modalities for the fractures were similar for both groups, as was the fracture final outcome. Local soft tissue injury was more severe in the gunshot injury group, as demonstrated by a higher number of type IIIC fractures, as well as more nerve injuries. CONCLUSION: Terror attacks may produce several modes of severe penetrating injuries causing high-grade open fractures. These should be aggressively treated by physicians remaining cognizant of other systemic and general implications of such a severe trauma. PMID- 17426548 TI - Trimed fixation system for displaced fractures of the distal radius. AB - OBJECTIVE: To study the functional outcome of displaced distal radius fractures treated by open reduction and internal fixation with the use of fragment-specific fixation. METHODS: Fifteen consecutive displaced distal radius fractures (5 extra articular and 10 intra-articular) were treated using the Trimed distal radius fixation system (Trimed, Valencia, CA). Radiographic assessment and range of motion of the affected wrist were evaluated postoperatively. Functional outcomes were evaluated with use of the Disabilities of the Arm, Shoulder, and Hand questionnaire and the Gartland and Werley scoring system. RESULTS: The mean follow-up was 11 months. The patients initiated controlled passive and active motion exercises during the first week. The mean score on the Disabilities of the Arm, Shoulder, and Hand questionnaire was 16 points. The mean return to work was 12 weeks. The functional outcome was 80% excellent or good by the Gartland and Werley scoring system for intra-articular fractures and 80% excellent for extra articular fractures. Range of active motion in intra-articular fractures was measured 60 degrees wrist extension, 64 degrees flexion, 65 degrees pronation, and 70 degrees supination, and for extra-articular fractures, 75 degrees extension, 70 degrees flexion, 80 degrees pronation, and 75 degrees supination. There was failure of fixation with loss of the reduction that was achieved at the time of operation in one patient. CONCLUSIONS: Fragment-specific fixation of unstable distal radius fractures using the Trimed system appears to be effective. Anatomic reduction, earlier range of motion, and improved functional results can be achieved using low-profile implants. PMID- 17426549 TI - Determinants of long-term functional consequences after major trauma. AB - OBJECTIVE: The purpose of this study was to describe the long-term functional consequences from major trauma and to quantify the effect of sociodemographic, injury-related, and physical determinants of its outcome. METHODS: A prospective cohort study was performed at the University Medical Center Utrecht (Level I trauma center) in the Netherlands during 1999 and 2000. All severely (injury severity score [ISS] >or=16) injured adult (age >or=16) trauma survivors (n = 359) were selected for follow-up. Between 12 and 18 months after trauma, outcome was assessed by means of Glasgow Outcome Scale (GOS), EuroQol (EQ-5D), and cognitive complaints. RESULTS: Follow-up assessments (overall response rate 93%) were obtained of 335 patients (249 men, 86 women) with a mean age of 38 years (SD = 17) and a mean ISS of 25 (SD = 10.6). The mean visual analog scale score on the EuroQol (EQvas) was 73.5 (SD = 17.8) and the mean utility score (EQus) was 69.1 (SD = 29.9), both below the norm. Patients reported limitations of mobility (48%); self-care (18%); daily activities (55%); pain and discomfort (63%); anxiety or depression (28%); and cognitive complaints (65%). In multivariate analyses, injury localization (spinal cord injury, lower extremity injury, or brain injury) was significantly associated with EQvas, EQus, and other outcome measures. Educational level was significantly associated with EQvas, anxiety/depression, and cognitive complaints. Comorbidity was significantly associated with EQvas, EQus, all dimensions of the EQ-5D (except anxiety/depression), and cognitive complaints. CONCLUSION: In addition to the injury localization, educational level and comorbidity were identified as important independent predictors of long-term functional consequences after major trauma. These determinants need further attention in outcome research and clinical practice. PMID- 17426550 TI - Hemodynamic patterns preceding circulatory deterioration and death after trauma. AB - OBJECTIVE: To describe the sequence of hemodynamic changes associated with sudden circulatory deterioration compared with those of terminal patients to identify the earliest signs warning of shock and death. METHODS: This is a prospective observational study of 89 patients with thermodilution cardiac index and continuous noninvasive hemodynamic monitoring who had episodes of circulatory deterioration. These data were compared with the data of a second group of 24 patients in their terminal stage just before death. RESULTS: The earliest indications of impending collapse were decreased cardiac index, and tissue perfusion reflected by decreased transcutaneous O2 tension (PtcO2). This was followed by reduced blood pressure, tachycardia, reduced arterial hemoglobin saturation, and increased transcutaneous CO2 tension (PtcCO2). This pattern of changes was more pronounced in the nonsurvivors and was seen in exaggerated form in terminal patients. CONCLUSION: Sequential hemodynamic patterns revealed reduced blood flow and poor tissue perfusion as the earliest warning signs in both circulatory deterioration and death. These were followed by reduced mean arterial pressure, tachycardia, and low values of pulse oximetry. Adequate blood flow and even distribution of flow are needed for tissue perfusion. PMID- 17426551 TI - Evidence-based validation of the predictive value of the American Association for the Surgery of Trauma kidney injury scale. AB - BACKGROUND: To evaluate the predictive value of the American Association for the Surgery of Trauma (AAST) kidney injury scale for the management of traumatic renal injuries. METHODS: From October 1995 through October 2004, 424 patients presented to our hospital with traumatic renal injury. RESULTS: Overall, 27.8% of patients had grade I injury, 26.4% had grade II injury, 19.3% had grade III injury, 18.2% had grade IV injury, and 8.3% had grade V injury. Patient age, Glasgow Coma Scale score, Revised Trauma Score, creatinine, blood urea nitrogen (BUN), white blood count, gender, substance abuse, shock, flank ecchymosis, abdominal pain, and mortality were not associated with AAST grade. Systolic blood pressure and hematocrit levels decreased with increasing AAST grades (p = 0.032 and p = 0.045, respectively). Volume transfused and length of hospitalization increased with AAST grades (p = 0.003 and p = 0.004, respectively). Patients with gunshot injury had higher AAST grades than those with blunt trauma (p < 0.001). Hypotension (14%), blood transfusion (47%), gross hematuria (65.9%), and flank pain (25%) were associated with higher AAST grades (p = 0.010, p < 0.001, p = 0.016, and p = 0.001, respectively). Ninety patients (21.2%) underwent renal exploration: 61% nephrectomies and 39% renorraphies. In multivariable analyses, type of injury, hematuria at presentation, and AAST scale predicted the risk of renal exploration (p < 0.001, p = 0.024, and p < 0.001, respectively), whereas type of injury and AAST scale were the sole predictors of nephrectomy (p < 0.001 and p < 0.001, respectively). CONCLUSIONS: We confirmed that the AAST injury severity scale is a powerful and valid tool for prediction of clinical outcome in patients with renal trauma. PMID- 17426552 TI - Predictive model for estimating risk of crush syndrome: a data mining approach. AB - BACKGROUND: There is no standard triage method for earthquake victims with crush injuries because of a scarcity of epidemiologic and quantitative data. We conducted a retrospective cohort study to develop predictive models based on clinical data for crush injury in the Kobe earthquake. METHODS: The medical records of 372 patients with crush injuries from the Kobe earthquake were retrospectively analyzed. Twenty-one risk factors were assessed with logistic regression analysis for three outcomes relating to crush syndrome. Two types of predictive triage models--initial evaluation in the field and secondary assessment at the hospital--were developed using logistic regression analysis. Classification accuracy, Brier score and area under the receiver operating characteristic curve (AUC) were used to evaluate the model. RESULTS: The initial triage model, which includes pulse rate, delayed rescue, and abnormal urine color, has an AUC of 0.73. The secondary model, which includes WBC, tachycardia, abnormal urine color, and hyperkalemia, shows an AUC of 0.76. CONCLUSIONS: These triage models may be especially useful to nondisaster experts for distinguishing earthquake victims at high risk of severe crush syndrome from those at lower risk. Application of the model may allow relief workers to better utilize limited medical and transportation resources in the aftermath of a disaster. PMID- 17426553 TI - Usefulness of the abbreviated injury score and the injury severity score in comparison to the Glasgow Coma Scale in predicting outcome after traumatic brain injury. AB - BACKGROUND: Assessment of injury severity is important in the management of patients with brain trauma. We aimed to analyze the usefulness of the head abbreviated injury score (AIS), the injury severity score (ISS), and the Glasgow Coma Scale (GCS) as measures of injury severity and predictors of outcome after traumatic brain injury (TBI). METHODS: Data were prospectively collected from 410 patients with TBI. AIS, ISS, and GCS were recorded at admission. Subjects' outcomes after TBI were measured using the Glasgow Outcome Scale (GOS-E) at 12 months postinjury. Uni- and multivariate analyses were performed. RESULTS: Outcome information was obtained from 270 patients (66%). ISS was the best predictor of GOS-E (rs = -0.341, p < 0.001), followed by GCS score (rs = 0.227, p < 0.001), and head AIS (rs = -0.222, p < 0.001). When considered in combination, GCS score and ISS modestly improved the correlation with GOS-E (R = 0.335, p < 0.001). The combination of GCS score and head AIS had a similar effect (R = 0.275, p < 0.001). Correlations were stronger from patients 8). CONCLUSIONS: GCS score, AIS, and ISS are weakly correlated with 12-month outcome. However, anatomic measures modestly outperform GCS as predictors of GOS-E. The combination of GCS and AIS/ISS correlate with outcome better than do any of the three measures alone. Results support the addition of anatomic measures such as AIS and ISS in clinical studies of TBI. Additionally, most of the variance in outcome is not accounted for by currently available measures of injury severity. PMID- 17426554 TI - A crisis in the delivery of care to patients with brain injuries in South Texas. AB - BACKGROUND: To determine the opinions of neurosurgeons regarding the care of the injured and to assess the impact of these attitudes on the care of the patients with brain injuries. METHODS: A survey was sent to the 2,465 active members of the American Association of Neurologic Surgeons. A manpower assessment of neurosurgical coverage of South Texas was also performed. RESULTS: In total, 872 surveys were returned (35%). Seventy-one percent of the respondents were over the age of 44. Eighty-seven percent of neurosurgeons stated that they currently provide trauma care: 74% at Level I or II trauma centers. The majority of neurosurgeons treated <5 trauma patients per week, 80% placed 2 or fewer intracranial pressure (ICP) monitors per month. Fifty-nine percent of the respondents preferred not to treat trauma patients because of (1) perceived increased medicolegal risk (80%), (2) conflict with elective practice (75%), (3) time required (70%), and (4) inadequate compensation (65%). Fifty-six percent received no compensation for trauma call. The majority of neurosurgeons indicated that no personnel other than neurosurgeons should be allowed to perform trauma craniotomies (90%) or insert ICP monitors (76%). However, 61% thought that non neurosurgeons should be able to perform neuro-critical care. A maldistribution of neurosurgeons was identified in South Texas, with much of the population uncovered for trauma care. Significant delays in definitive neurosurgical care were identified as a result of this maldistribution. CONCLUSIONS: One-half of neurosurgeons prefer not to care for trauma patients because of perceived added time commitment, conflicts with elective practice, lack of compensation, and perceived medicolegal risk. But, they thought that only neurosurgeons should provide emergency neurosurgical procedures. These attitudes appear to impinge on the care of the patients with brain injuries in South Texas. PMID- 17426555 TI - Predicting trauma inpatient mortality in an administrative database: an investigation of survival risk ratios using New York data. AB - BACKGROUND: A precise and unbiased measure of injury severity, calculable from administrative data sets, is needed for population-based studies. METHODS: The 2001 Nationwide Inpatient Sample was used to calculate independent and traditional survival risk ratios. Logistic regression models predicting survival for New York State's Statewide Planning and Research Cooperative System 2003 inpatients (117,630 records) were constructed for both types of survival risk ratios. The contribution of only the worst survival risk ratio (SRR), the two worst SRRs, the three worst, the four worst, and all trauma diagnoses was investigated. The contribution of gender and age to the models was assessed. RESULTS: Use of the two worst SRRs, gender, and age gives the best prediction of inhospital mortality. CONCLUSION: Inhospital mortality in both trauma centers and in community hospitals can be predicted well from an administrative database by using three predictors--gender, age, and the product of the SRRs for the two diagnoses with the lowest chance of survival. PMID- 17426556 TI - The reasons families donate organs for transplantation: implications for policy and practice. AB - BACKGROUND: The objective of this study is to examine the reasoning behind donation decisions of donor-eligible patients' next-of-kin. Cases were identified through chart review at hospitals in Ohio and Pennsylvania. Family decision makers of donor-eligible patients (N = 420) were interviewed to understand decisions regarding organ donation. METHODS: Responses to inquiries regarding donation decisions were recorded verbatim and then categorized into major themes. Patterns of reasons to donate or not were examined to identify trends. Logistic regression was employed to explore the associations between decision-maker characteristics and donation decision. RESULTS: Most families gave more than one reason for their decision. Respondents who chose to donate most frequently mentioned altruistic reasons for their decision (78%). However, this response was almost never provided as a single reason. Seventy-five percent of families attributed the decision to the patient's explicit communication of a desire to donate. The belief that the patient did not want to donate was cited by 51% of families who did not donate; 36% of these families cited this as the sole reason for not donating. Concerns about disfigurement and feelings of emotional exhaustion also played a significant role. CONCLUSIONS: Reasons to donate or not are complex. Altruism, although important, does not seem to be sufficient to motivate organ donation. Knowing patient's preferences was a stronger influence overall. Moreover, certain reasons were highly correlated with each other. Supportive emotional care for the family and reassurance and information about the donation process are indicated as important issues to address to encourage donation. PMID- 17426557 TI - Intrahepatic gas at postmortem computed tomography: forensic experience as a potential guide for in vivo trauma imaging. AB - BACKGROUND: Until August 2004 there were 106 forensic cases examined with postmortem multislice computed tomography (MSCT) and magnetic resonance (MR) imaging before traditional autopsy within the Virtopsy project. Intrahepatic gas (IHG) was a frequent finding in postmortem MSCT examinations. The aim of this study was to investigate its cause and significance. METHODS: There were 84 virtopsy cases retrospectively investigated concerning the occurrence, location, and volume of IHG in postmortem MSCT imaging (1.25 mm collimation, 1.25 mm thickness). We assessed and noted the occurrence of intestinal distention, putrefaction, and systemic gas embolisms and the cause of death, possible open trauma, possible artificial respiration, and the postmortem interval. We investigated the relations between the findings using the contingency table (chi2 test) and the comparison of the postmortem intervals in both groups was performed using the t test in 79 nonputrefied corpses. RESULTS: IHG was found in 47 cases (59.5%). In five of the cases, the IHG was caused or influenced by putrefaction. Gas distribution within the liver of the remaining 42 cases was as follows: hepatic arteries in 21 cases, hepatic veins in 35 cases, and portal vein branches in 13 cases; among which combinations also occurred in 20 cases. The presence of IHG was strongly related to open trauma with systemic gas. Pulmonary barotrauma as occurring under artificial respiration or in drowning also caused IHG. Putrefaction did not seem to influence the occurrence of IHG until macroscopic signs of putrefaction were noticeable. CONCLUSIONS: IHG is a frequent finding in traumatic causes of death and requires a systemic gas embolism. Exceptions are putrefied or burned corpses. Common clinical causes such as necrotic bowel diseases appear rarely as a cause of IHG in our forensic case material. PMID- 17426558 TI - Analysis of injury- and violence-related fatalities in the Ohio Medicaid population: identifying opportunities for prevention. AB - BACKGROUND: To identify the leading causes of injury- and violence-related deaths in demographic subgroups of the population in Ohio, by Medicaid status. METHODS: We used linked Ohio Medicaid and death certificate files, 1992 to 1998, and obtained the probability (p) of dying from a specific mechanism of injury--given death from injury--by Medicaid status, using multinomial multivariable logistic regression analysis. Probabilities were rank-ordered to identify the leading causes of death in each subgroup. RESULTS: The leading cause of injury-related deaths was homicide among Medicaid decedents in the age groups 0 to 4, 15 to 24, and 25 to 44 (p = 0.283, 0.380, and 0.269, respectively), and motor vehicle crashes among nonMedicaid decedents aged 5 to 14, 15 to 24, 25 to 44, and 45 to 74 (p = 0.448, 0.462, 0.293, and 0.293, respectively). Accidental falls ranked first among the elderly (p = 0.593 and 0.414, respectively in Medicaid and nonMedicaid decedents). Suicide and accidental exposure to smoke, fire, and flames also ranked high among the leading causes of injury-related deaths in many population subgroups. CONCLUSIONS: Findings from this study, pointing to the vulnerability of population subgroups to certain mechanisms of injury, can be used to formulate targeted prevention strategies. PMID- 17426559 TI - Risk-inducing activities leading to injuries in a child and adolescent population of Germany. AB - BACKGROUND: Injuries are the most common cause of mortality in children, also accounting considerably for childhood morbidity. However, data on injuries only provide valid information on the actual risk of each injury-causing activity when taken in consideration of the relationship with actual activity exposure data. Therefore, the primary goal of this investigation is to determine the relative risk of normal child and adolescent activities. METHODS: From January 1, 1999 to December 31, 2001, a school questioning in regard to social, pedagogic, and leisure activities was performed among 2,325 students ranging from 6 to 17 years old. A total of 3,645 injuries sustained by children and adolescents treated at the surgical emergency department of the University Hospital Dresden were analyzed. Furthermore, a danger awareness test was performed. RESULTS: Forty three percent of all injuries happened during leisure time, 41% at school, 8% in traffic, and 8% at home. Bicycle riding was pointed out as the most frequent leisure activity, regardless of gender and age. Horse riding had a 9-fold increased risk and moped driving had a 23.75-fold increased risk for injury compared with adolescent bike riding. Horse riding and snowboarding showed an increased risk for injury in children (5.6- and 4.2-fold, relative to biking). The level of danger awareness was significantly lower in children with a history of frequent injuries. CONCLUSIONS: The riskier activities were horse-riding, moped driving, and snowboarding. The level of danger awareness did affect the frequency of injuries. The authors recommend a danger awareness test for all children to identify those who would benefit from injury prevention training. PMID- 17426560 TI - Evaluation of the relationship between mechanism of injury and outcome in pediatric trauma. AB - BACKGROUND: Most prehospital triage strategies are based on physiologic, anatomic, and mechanism-related variables. Although previous studies have suggested the value of physiologic and anatomic triage criteria, the predictive capacity of mechanism of injury has been questioned. The purpose of the current study was to evaluate the relationship between mechanism of injury and resource utilization and outcome among injured children treated at trauma centers. METHODS: The relationship between mechanism of injury and mortality and resource utilization (need for operative care, total and ICU length of stay) was analyzed using the records of pediatric patients (age <15 years) included in the National Pediatric Trauma Registry between 1995 and 2001. RESULTS: Significant variability in the outcome, resources requirements, and need for inpatient rehabilitation after discharge were observed among the mechanisms analyzed. Mechanisms such as firearm injuries were more likely to be severe and require significant trauma center resources, whereas other mechanisms such as falls related to stairs were more likely to result in injuries that were less severe and require relatively few resources. A proposed framework is presented into which mechanisms are stratified according to severity of injury (high vs. low severity) and need for trauma center resources (high vs. low requirement). CONCLUSIONS: Mechanism of injury is associated with the need for trauma center care but this association is highly dependent on the measure used to determine appropriateness of triage. PMID- 17426561 TI - The consequences of high-risk behaviors: trauma during pregnancy. AB - BACKGROUND: Trauma during pregnancy places two lives at risk. Knowledge of risk factors for trauma during pregnancy may improve outcomes. METHODS: We reviewed the charts of 188 such patients admitted to a Level I trauma center from 1996 to 2004. A comparison was made of injury severity and outcome from a cohort of nonpregnant female trauma patients selected with a similar temporal occurrence and age range. RESULTS: Motor vehicle collisions comprised 160 cases, 67 using a restraint device. Of 84 patients tested, 45 tested positive for intoxicants, 16 positive for 2 or more intoxicants. A significant trend toward less testing through the study period was observed (p = 0.0002). Injury severity was assessed by Revised Trauma Score (RTS). RTS <11 or admission to operating room or intensive care units (OR/ICU) classified patients as severely injured. The six maternal fatalities had an RTS <11 or OR/ICU disposition. Fetal outcomes included 155 live in utero, 18 live births, and 15 fatalities correlating with injury severity by either criteria (p < 0.0001). Of the fetal fatalities, 7 occurred with RTS = 12, but only 3 fatalities occurred in the 147 cases not admitted to OR/ICU. Gestational age correlated (p < 0.0001) with fetal outcomes. The 18 live births had mean gestational ages of 35 +/- 4 weeks as compared with fetal fatalities at 20 +/- 9 weeks, and fetuses alive in utero at 22 +/- 9 weeks gestation. Coagulation tests prothrombin time (PT), international normalized ratio (INR) (both p < 0.008), and partial thromboplastin time (PTT) (p < 0.0001) correlated with maternal outcome. A matched cohort of nonpregnancy trauma cases during the same time frame indicated that, despite a significantly higher percentage of severely injured patients, fewer fatalities occurred. This might reflect a greater risk for the pregnant trauma patient. CONCLUSIONS: This study of trauma in pregnancy cases revealed a high percentage with risk behaviors. There was a significant trend toward less intoxicant testing in recent years. Coagulation tests were the most predictive of outcomes. Lower gestational age correlated with fetal demise. PMID- 17426562 TI - Attempted suicide and the elderly trauma patient: risk factors and outcomes. AB - OBJECTIVE: To analyze the scope and burden of hospitalizations for suicide attempts among elderly patients in the United States. METHODS: The National Trauma Data Bank (NTDB, American College of Surgeons, Chicago, IL, 2002) was used for this study. It is a multistate database of hospitalizations for traumatic injury in the United States. Information on all patients reported to the database from 1995 to 2002 was analyzed. Logistic regression was used to analyze the risk factors for suicide attempt in elderly patients, compared with both a younger suicidal cohort and a cohort of elderly patients injured in a motor vehicle collision (MVC). The impact of age on outcome after failed suicide attempt was also analyzed. RESULTS: A total of 1,812 persons aged 65 and over were hospitalized for suicide attempts during the study period, comprising 9.5% of total hospitalizations for suicide attempts. Regression analysis demonstrated that elderly patients who attempted suicide were more likely to be male, white, to have used a firearm, and to have insurance than younger patients. They were more likely to have a psychiatric condition but less likely to have insurance than elderly patients hospitalized for MVCs. Mortality was higher for elderly patients hospitalized for suicide attempts than for younger patients who attempted suicide. Suicidal elderly were less likely to be discharged to home than either younger suicidal patients or elderly patients hospitalized after MVCs. CONCLUSIONS: The failed suicide attempt is an opportunity for intervention. By better understanding risk factors and outcomes of suicide attempts among elderly patients, we can identify higher risk groups and begin to tailor social service programs, psychiatric interventions, and medical care. A multimodality approach to suicide prevention for the elderly would include compassionate, appropriate, psychosocial interventions, and could be studied prospectively to analyze its impact. PMID- 17426563 TI - Household firearm ownership and rates of suicide across the 50 United States. AB - BACKGROUND: The current investigation explores the association between rates of household firearm ownership and suicide across the 50 states. Prior ecologic research on the relationship between firearm prevalence and suicide has been criticized for using problematic proxy-based, rather than survey-based, estimates of firearm prevalence and for failing to control for potential psychological risk factors for suicide. We address these two criticisms by using recently available state-level survey-based estimates of household firearm ownership, serious mental illness, and alcohol/illicit substance use and dependence. METHODS: Negative binomial regression was used to assess the relationship between household firearm ownership rates and rates of firearm, nonfirearm, and overall suicide for both sexes and for four age groups. Analyses controlled for rates of poverty, urbanization, unemployment, mental illness, and drug and alcohol dependence and abuse. RESULTS: US residents of all ages and both sexes are more likely to die from suicide when they live in areas where more households contain firearms. A positive and significant association exists between levels of household firearm ownership and rates of firearm and overall suicide; rates of nonfirearm suicide were not associated with levels of household firearm ownership. CONCLUSION: Household firearm ownership levels are strongly associated with higher rates of suicide, consistent with the hypothesis that the availability of lethal means increases the rate of completed suicide. PMID- 17426564 TI - Traumatic thoracic aortic pseudocoarctation treated with an endograft. PMID- 17426565 TI - Traumatic abdominal aortic dissection in a child. PMID- 17426566 TI - Survival after simultaneous blunt injury to the ascending aorta and transverse arch. PMID- 17426567 TI - Endoscopic management of traumatic hemobilia. PMID- 17426568 TI - Late presentation of a hepatic pseudoaneurysm with hemobilia after angioembolization for blunt hepatic trauma. PMID- 17426569 TI - A neglected retained penrose drain mimicking an amputation stump neuroma. PMID- 17426570 TI - Iatrogenic arteriovenous fistula after trans-heel traction pin: a case report. PMID- 17426571 TI - Sir Ivan Magill: the right physician in the right place at the right time. PMID- 17426572 TI - Soft palatal injury resulting from an unusual fishhook in a child. PMID- 17426573 TI - Guidelines for prevention, diagnosis, and treatment of ventilator-associated pneumonia (VAP) in the trauma patient. PMID- 17426574 TI - Interpersonal violence and motor vehicle crashes caused the most significant amount of maxillofacial trauma. PMID- 17426576 TI - Evaluating anemia as a risk factor for worse neurologic outcome after traumatic brain injury (TBI). PMID- 17426577 TI - Anemia management program reduces transfusion volumes, incidence of ventilator associated pneumonia, and cost in trauma patients. PMID- 17426580 TI - Survey study estimating surge capacity in trauma centers in the United States. PMID- 17426582 TI - Splenic artery embolization: have we gone too far? PMID- 17426583 TI - Dosing of solvent/detergent plasma versus single-donor fresh-frozen plasma. PMID- 17426584 TI - Non-invasive hemodynamic monitoring using USCOM in HEMS at the scene. PMID- 17426587 TI - Cation Cl- cotransporters in the dendrites of goldfish bipolar cells. AB - Bipolar cells receive an input from gamma-aminobutyric acid ergic horizontal cells in distal retinas. gamma-Aminobutyric acid can induce either an inhibitory or an excitatory Cl(-) response based on the intracellular Cl(-) concentration in the bipolar dendrites. Two cation Cl(-) cotransporters, Na-K-2Cl(-) and K-2Cl(-), differently control local Cl(-) levels in neurons by uptaking or extracting Cl( ), respectively. By using immunocytochemical techniques, we found that Na-K-2Cl( ) and K-2Cl(-) were preferentially distributed in On-bipolar and Off-bipolar dendrites in goldfish retinas. Na-K-2Cl(-) was colocalized with protein kinase C in On-bipolar dendrites, whereas K-2Cl(-) was located in Off-bipolar dendrites, fully overlapping with ionotropic glutamate receptor GluR4. Possibly, an internal Cl(-) level is higher in the On-bipolar dendrites than in the Off-bipolar dendrites. gamma-Aminobutyric acid inputs in the distal retina might excite On bipolar cells, but inhibit Off-bipolar cells. PMID- 17426588 TI - An event-related potential study on the early processing of crying faces. AB - Crying is an attachment behavior, which in course of evolution had survival value. This study examined the characteristics of the face-sensitive N170, and focused on whether crying expressions evoked different early event-related potential waveforms than other facial expressions. Twenty-five participants viewed photographs of six facial expressions, including crying, and performed an implicit processing task. All stimuli evoked the N170, but the facial expression modulated this component in terms of latency and amplitude to some extent. The event-related potential correlates for crying faces differed mostly from those for neutral and fear faces. The results suggest that facial expressions are processed automatically and rapidly. The strong behavioral and emotional responses to crying appear not to be reflected in the early brain processes of face recognition. PMID- 17426589 TI - Functional magnetic resonance imaging of verbal learning in children with heavy prenatal alcohol exposure. AB - We examined functional MRI activation patterns corresponding to verbal paired associate learning in a group of 11 children with heavy prenatal alcohol exposure compared with 16 typically developing children. Among the typically developing children, prominent activation was observed in the left medial temporal lobe, left dorsal frontal lobe and bilateral posterior temporal cortices during learning and recall. Analyses revealed significantly less activation in left medial and posterior temporal regions and significantly more activation in right dorsal frontal cortex in the alcohol-exposed children relative to controls, even when group differences in memory test performance were statistically controlled. These results may indicate an increased reliance on frontal memory systems in the children with heavy prenatal alcohol exposure, perhaps compensating for dysfunctional medial temporal memory systems. Our findings are consistent with neuropsychological and structural imaging studies, and provide the first evidence for brain activation abnormalities, independent of group performance differences, during verbal learning and recall in children with heavy prenatal alcohol exposure. PMID- 17426590 TI - Musashi-1 expression in postnatal mouse olfactory epithelium. AB - We investigated the age-related change in the distribution of a molecular marker for neural stem and precursor cells, Musashi-1, in the olfactory epithelium of mice from 1 day up to 16 months of age using immunohistochemistry. We also compared the distribution pattern of Musashi-1 with that of growth-associated protein 43, the olfactory marker protein, and Notch-1. Musashi-1 was expressed in the globose basal cell layer and the lower part of the growth-associated protein 43-positive layer, with immunoreactivity decreasing with aging. Notch-1 was observed only in the early postnatal period. These findings are consistent with the fact that globose basal cells are proliferating olfactory precursor cells and that their ability to generate new neurons decreases with aging. PMID- 17426591 TI - Vibrotactile activation of the auditory cortices in deaf versus hearing adults. AB - Neuroplastic changes in auditory cortex as a result of lifelong perceptual experience were investigated. Adults with early-onset deafness and long-term hearing aid experience were hypothesized to have undergone auditory cortex plasticity due to somatosensory stimulation. Vibrations were presented on the hand of deaf and normal-hearing participants during functional MRI. Vibration stimuli were derived from speech or were a fixed frequency. Higher, more widespread activity was observed within auditory cortical regions of the deaf participants for both stimulus types. Life-long somatosensory stimulation due to hearing aid use could explain the greater activity observed with deaf participants. PMID- 17426592 TI - Somatosensory pain does not affect total cerebral blood volume. AB - The effect of somatosensory pain on the total cerebral blood volume was investigated in anesthetized rats. Our results show for the first time that total cerebral blood volume remains unaltered in both brain hemispheres during 2.5 min noxious stimulation of the sensory C-fibres of the sciatic nerve. Regional cerebral blood flow was increased by 97% in the thalamus and by 47% in the hypothalamus at the same time. Blockade of the L-arginine-nitric oxide system reduced significantly the steady-state control level of total cerebral blood volume (i.l.: from 5.7+/-1.3 to 4.58+/-1.6 vol%, c.l.: from 5.0+/-0.6 to 4.24+/ 0.9 vol%). Nitric oxide synthase blockade, however, did not affect either the stimulation induced increase of regional cerebral blood flow or the steadiness of total cerebral blood volume during the stimulation. PMID- 17426593 TI - If-then planning modulates the P300 in children with attention deficit hyperactivity disorder. AB - Children with attention deficit hyperactivity disorder have difficulties with tasks that require response inhibition. We measured electroencephalographic data of nonmedicated children with attention deficit hyperactivity disorder and control children in two conditions: (a) a neutral condition without a self regulation strategy and (b) a condition that involved the making of if-then plans (i.e. 'If situation X is encountered, then I will perform the goal-directed behavior Y'). If-then plans improved response inhibition and increased the P300 in children with attention deficit hyperactivity disorder compared with the neutral condition. The present results encourage the application of self regulation using if-then plans in addition or as an alternative to common medical therapy. PMID- 17426594 TI - Dopamine D2 receptor stimulation promotes the proliferation of neural progenitor cells in adult mouse hippocampus. AB - We initially examined the effects of apomorphine in vitro using mouse embryonic and adult neural progenitor cells. The effects of apomorphine treatment led to dose-dependent increases in the number of embryonic and adult neural progenitor cells, and dopamine D2 receptor antagonist treatment significantly reduced the increases induced by apomorphine. Next, we investigated the effects of apomorphine in vivo in the adult mouse hippocampus. The effects of single-dose apomorphine administration led to an increase of approximately 30% in the number of bromodeoxyuridine-positive cells in the dentate gyrus. Moreover, the chronic apomorphine administration induced an increase in the number of bromodeoxyuridine positive cells by about 30%. Thus, we suggest that the stimulation of dopamine D2 receptors increases the proliferation of neural progenitor cells both in vivo and in vitro. PMID- 17426595 TI - Somatotopy of corticospinal tract in the internal capsule shown by functional MRI and diffusion tensor images. AB - Using functional MRI and diffusion tensor tractography, we studied the topographical relation of hand and foot fibers of the corticospinal tract within the internal capsule to verify the recent unexpected finding by Holodny et al., who reported that hand fibers are located anterolateral to foot fibers, not anteromedial as is currently believed. The location of hand fibers with respect to foot fibers was anterolateral in four participants, posterolateral in two, and anteromedial in one of seven participants examined. Thus, there was some support for the anterolateral finding of Holodny et al., but interindividual variability was also indicated. PMID- 17426596 TI - Galectin-3 inhibits Schwann cell proliferation in cultured sciatic nerve. AB - The production of galectin-3, a carbohydrate-binding mammalian lectin, is upregulated in Schwann cells after peripheral nerve injury in areas where Schwann cells proliferate. Here we tested if galectin-3 affected proliferation of Schwann cells in cultured sciatic nerve segments. Galectin-3 significantly decreased the number of bromodeoxyuridine-labelled Schwann cell nuclei. Neither lactose nor a synthetic inhibitor directed against the carbohydrate-binding region abolished the effects of galectin-3. In addition, a mutant galectin-3 unable to bind endogenous carbohydrates had similar effects as normal galectin-3. We conclude that galectin-3 reduces proliferation of Schwann cells in cultured sciatic nerve segments by a mechanism which is independent of its carbohydrate-binding moiety. PMID- 17426597 TI - Cardiovascular response to renin substrate microinjection into the central nucleus of the amygdala of rats. AB - Central nucleus of the amygdala is involved in cardiovascular regulation. Although most components of the renin-angiotensin system have been found to be distributed in amygdala, renin expression in brain has remained controversial. This work was undertaken to elucidate the extent of renin presence in this nucleus. A cannula was implanted bilaterally into the central nucleus of the amygdala. Mean arterial pressure and heart rate were directly measured via indwelling femoral artery cannula post bilateral intra central nucleus of the amygdala microinjection of renin substrate. Renin substrate microinjection dose dependently increased mean arterial pressure and heart rate, whereas captopril, saralasin and losartan pretreatment inhibited these effects. The results suggest the presence of local renin or similar proteases in this nucleus. PMID- 17426598 TI - Water avoidance stress activates colonic myenteric neurons in female rats. AB - Stress stimulates colonic motor function and plays a role in functional bowel disorders, prevalently in women. We examined, in conscious female rats, the influence of water avoidance stress for 60 min on colonic myenteric neuron activity using immunohistochemical detection of Fos as a marker of neuronal activity. In control rats, Fos immunoreactive nuclei were rare in proximal and distal colon and no defecation was observed. Water avoidance stimulated fecal pellet output, which was associated with Fos expression in myenteric ganglia of proximal and distal colon including in a population of peripheral choline acetyltransferase-immunoreactive neurons. Atropine blocked fecal pellet output but not Fos expression in myenteric ganglia. These results indicate that psychological stress stimulates the activity of colonic cholinergic myenteric neurons. PMID- 17426599 TI - Neural correlates of stimulus-reward association in the rat mediodorsal thalamus. AB - Neural activity was recorded from the mediodorsal thalamic nucleus of behaving rats during the discrimination of olfactory cues associated with or without a reward. Approximately 10% of mediodorsal thalamic neurons showed significant responses during the presentation of cues. Most of these neurons responded strongly to cues associated with a reward. These neurons also differentiated between cues, even in the same reinforcement (reward/nonreward) group. All of the neurons tested in extinction, relearning, and/or new-learning trials changed their responses flexibly according to the reward contingency. These neurons were located primarily in the central and medial segments of the mediodorsal thalamus. These results suggest that these mediodorsal thalamic neurons are the neural substrates for association learning of olfactory stimuli with rewards. PMID- 17426600 TI - Involvement of M1-muscarinic acetylcholine receptors, protein kinase C and mitogen-activated protein kinase in the effect of huperzine A on secretory amyloid precursor protein-alpha. AB - This study is to explore the involvement of muscarinic acetylcholine receptors/protein kinase C cascade and the mitogen-activated protein kinase pathway in the effect of huperzine A on the secretory amyloid precursor protein alpha. Upregulation of secretory amyloid precursor protein-alpha by huperzine A was attenuated by muscarinic acetylcholine receptor antagonist (specifically by M1-muscarinic acetylcholine receptor antagonist), and markedly blocked (-37.7%) by protein kinase C inhibitor as well. Meanwhile, huperzine A can activate the phosphorylation of mitogen-activated protein kinase and, accordingly, partly restored PD98059-decreased secretory amyloid precursor protein-alpha secretion. In addition, huperzine A largely inhibited (-55.4%) acetylcholinesterase activity of the cell line. Our results suggest that activated M1-muscarinic acetylcholine receptor/protein kinase C pathway and mitogen-activated protein kinase signaling are involved in the process of huperzine A enhancing the secretory amyloid precursor protein-alpha secretion. PMID- 17426601 TI - Are you looking at me? Neural correlates of gaze adaptation. AB - Eye gaze is an important social signal, and humans can accurately determine gaze direction in others. Recently, dramatic adaptation effects on gaze perception were shown, in which the perception of small gaze deviations to the adapted direction is diminished. We compared participants' perceptions of gaze direction before and after adaptation to the left or right gaze, and examined event-related potential correlates of this gaze adaptation. We found a striking impairment in perceiving gaze to the adapted side. Event-related potentials revealed no direction-specific gaze adaptation effects on N170, although small adaptation effects were seen later, approximately 250-350 ms. This suggests that, rather than modulating the gaze processing approximately 170 ms in posterior occipitotemporal areas, adaptation modulates subsequent processes that are possibly mediated by more anterior right-temporal areas. PMID- 17426602 TI - Cortical activity during the recognition of cooperative actions. AB - We investigated brain activity during the recognition of cooperative actions between two persons. Participants were requested to observe movie clips in which one person received an object passed by another person and to rate the appropriateness of the hand-motion patterns of the receiver. The relative oxyhaemoglobin concentrations in the regions that are considered to be involved in the mirror neuron system (the inferior frontal and precentral areas) were measured by 24-channel near-infrared spectroscopy. The results revealed that the relative oxyhaemoglobin concentration increased to a greater extent during the observation of appropriate receiving actions than during the observation of inappropriate ones. These results suggest that the mirror neuron system is involved in the comprehension of cooperative actions. PMID- 17426603 TI - Prediction of relative and absolute time of reward in monkey prefrontal neurons. AB - We studied single-neuron activity in the monkey dorsolateral prefrontal cortex during a saccade task, in which correct responses were rewarded after a delay of 0.5 or 1.5 s in one trial-block, and after 1.5 or 3-s delay in the other trial block. Activity of some neurons depended on the relative length of the delays (longer or shorter) within each block, and activity for the 1.5-s trials was significantly different between the blocks. Activity of another group of neurons reflected the absolute length of delay: hence, the activity in the 1.5-s trials did not differ between the blocks. These results indicate that both relative and absolute time of future reward is represented in subsets of neurons in the dorsolateral prefrontal cortex. PMID- 17426604 TI - Effects of meditation on frontal alpha-asymmetry in previously suicidal individuals. AB - This study investigated the effects of a meditation-based treatment for preventing relapse to depression, mindfulness-based cognitive therapy (MBCT), on prefrontal alpha-asymmetry in resting electroencephalogram (EEG), a biological indicator of affective style. Twenty-two individuals with a previous history of suicidal depression were randomly assigned to either MBCT (N=10) or treatment-as usual (TAU, N=12). Resting electroencephalogram was measured before and after an 8-week course of treatment. The TAU group showed a significant deterioration toward decreased relative left-frontal activation, indexing decreases in positive affective style, while there was no significant change in the MBCT group. The findings suggest that MBCT can help individuals at high risk for suicidal depression to retain a balanced pattern of baseline emotion-related brain activation. PMID- 17426605 TI - Transgenic glial nuclear factor-kappa B inhibition decreases formalin pain in mice. AB - In this work, we studied transgenic glial fibrillary acidic protein-IkappaBalpha dn mice that selectively inactivate the classical nuclear factor kappaB pathway by overexpressing the inhibitory protein of kappaBalpha in astrocytes, under the control of glial fibrillary acidic protein promoter. We sought to determine if glial nuclear factor kappaB inhibition decreases formalin pain. Formalin testing was carried out on 25-35 g littermate adult male wild-type and transgenic C57Bl/6 mice. Formalin increased spinal cord c-Fos expression and glial fibrillary acidic protein immunostaining in both wild-type and transgenic mice. Transgenic glial fibrillary acidic protein-inhibitory protein of kappaBalpha-dn mice had lower duration of formalin-induced paw-licking behavior. These data support a role of glial nuclear factor kappaB inhibition in reducing pain after peripheral nerve inflammation. PMID- 17426606 TI - Involvement of hippocampal metabotropic glutamate receptors in radial maze performance. AB - We investigated the role of hippocampal metabotropic glutamate receptors in spatial learning and memory, using an eight-arm radial maze task. (S)-4 Carboxyphenylglycine, a group I metabotropic glutamate receptor antagonist, or trans-(+/-)-1-amino-(1S,3R)-cyclopentanedicarboxylic acid, a broad-spectrum metabotropic glutamate receptor agonist, was administered into the dorsal hippocampus after rats had acquired the task. Both of these agents significantly impaired radial maze performance, suggesting a functional importance of hippocampal metabotropic glutamate receptors in spatial working memory. PMID- 17426608 TI - Statistical and nonstatistical significance: implications for health care researchers. AB - Quality improvement professionals have to decide whether a change has led to improvement. This is typically done through testing the statistical significance of the findings. In this article, we explore controversies surrounding statistical significance testing with attention to contemporary criticism of bad practice resulting from the misuse of statistical significance testing. Most statistical significance tests use tests (eg, F, chi) with known distributions with the P values used as the main evidence to evaluate whether tests are statistically significant. The primary conclusion of this article is that the P value alone as a measure of statistical significance does not give sufficient information about testing of hypotheses. When it is coupled with other measures, however, such as the point estimation of the effect size and the use of a confidence interval around it, the combination of these statistics can provide a more thorough explanation of statistical testing. This article offers recommendations for process improvement investigators as to when to appropriately apply and not to apply statistical significance testing. PMID- 17426609 TI - National hospital antibiotic timing measures for pneumonia and antibiotic overuse. AB - The development of drug-resistant bacteria from the overuse of antibiotics is a serious problem, with overutilization threatening to disarm caregivers and their patients even as together they face increasingly virulent strains of microbes. On the other hand, the speedy treatment of pneumonia with antibiotics is a firmly established, evidence-based practice, enshrined in Joint Commission on Accreditation for Healthcare Organizations Core Measures used in hospital accrediting and public reporting, and in Centers for Medicare and Medicaid Services (CMS) public-reporting and pay-for-performance hospital measures. This sets the stage for a potential conflict between (a) not doing the wrong thing by overprescribing antibiotics and (b) prescribing antibiotics on time for pneumonia. In November 2005, pneumonia antibiotic timing results were announced for the 133 top-performing hospitals in the first year of the 3-year CMS Hospital Quality Incentive Demonstration (HQID) pay-for-performance project, conducted in collaboration with Premier Inc, a hospital purchasing and informatics alliance. Premier client hospitals participating in the HQID also submit drug utilization and other comparative data to Premier for client access for benchmarking purposes; this makes it possible to see how the antibiotics specified for pneumonia are used by Premier hospitals for other conditions. In this study we look at where increased success in meeting the HQID pneumonia antibiotic timing measure is tied to an increase in antibiotic use for conditions where antibiotics are unwarranted--with the potential for promoting antibiotic resistance. PMID- 17426610 TI - Control limits for p control charts with small subgroup sizes. AB - The p chart is widely used in health care and other service organizations as well as in manufacturing to monitor the proportion of observations with some particular characteristic for comparing several sources of data or for tracking a single source of data over time. The conventional approach is to use 3sigma limits found by using the normal approximation to the binomial distribution. This article reviews a method for taking into account the fact that 3sigma limits are not always appropriate, and suggests the use of the exact binomial distribution instead of the normal approximation to eliminate the problems associated with small subgroups. An example with only 4 small subgroups shows that the use of probability control limits and of modified control limits avoids the above problems. PMID- 17426611 TI - A practical limit to trials needed in one-person randomized controlled experiments. AB - Recently in this journal, J. Olsson and colleagues suggested the use of factorial experimental designs to guide a patient's efforts to choose among multiple interventions. These authors argue that factorial design, where every possible combination of the interventions is tried, is superior to sequential trial and errors. Factorial design is efficient in identifying the effectiveness of interventions (factor effect). Most patients care only about feeling better and not why their conditions are improving. If the goal of the patient is to get better and not to estimate the factor effect, then no control groups are needed. In this article, we show a modification in the factorial design of experiments proposed by Olsson and colleagues where a full-factorial design is planned, but experimentation is stopped when the patient's condition improves. With this modification, the number of trials is radically fewer than those needed by factorial design. For example, a patient trying out 4 different interventions with a median probability of success of .50 is expected to need 2 trials before stopping the experimentation in comparison with 32 in a full-factorial design. PMID- 17426612 TI - Perception and use of guidelines and interprofessional dynamics: assessing their roles in guideline adherence in delivering medications in nursing homes. AB - PURPOSE: Focusing on guidelines governing medication use in nursing homes, in this study we explore the ways in which clinical professionals perceive and apply treatment guidelines, and how interprofessional interactions shape the delivery of pharmacotherapies to residents. DESIGN AND METHODS: Seventeen semistructured interviews were conducted with physicians, nursing staff, and consultant pharmacists in 4 purposefully selected nursing homes in Michigan. RESULTS: Perceptions of guidelines varied by clinical groups, with physicians perceiving them as reference tools, whereas nurses and consultant pharmacists saw them as rules to which clinical practices should adhere. Key external factors including resident and family demands, economic constraints, limited face-to-face contact, and the organizational hierarchy further drove clinical personnel to use guidelines differently and induced interprofessional conflicts. These negative interactions make facility-wide guideline adherence challenging by undermining the regulatory role of consultant pharmacists. IMPLICATIONS: This study provides important insight on work-related factors that hinder the implementation of treatment guidelines in nursing homes. PMID- 17426613 TI - The relationship between pediatric volume and information technology adoption in hospitals. AB - OBJECTIVE: Numerous studies have identified organizational factors related to the adoption of information technology (IT) by hospitals. However, no study has examined whether patient characteristics of hospitals are related to the adoption of health IT. This study examines IT adoption in hospitals that care for either a large number or a large proportion of children. METHODS: Primary data from an IT survey of acute care hospitals were combined with secondary data on hospital discharges. Pediatric volume was both categorically and continuously operationalized in several ways. IT adoption was examined both at the individual IT application level and in several aggregate measures of organizational adoption. Univariate and linear regression models were used to analyze the data. Regression models controlled for average patient severity of illness (case mix), public insurance volume, bed size, and system affiliation for each hospital. RESULTS: All 98 acute care hospitals that participated in the survey were matched to the hospital discharge data. Analyses suggest that IT adoption is positively correlated with a higher absolute number of pediatric discharges from hospitals. Similarly, as children make up a higher percentage of a given hospital's discharges, the propensity to adopt clinical and nonclinical IT applications increases significantly as well. CONCLUSION: Acute care hospitals caring for a large number, or a large proportion, of children are more likely to adopt health IT. This relationship may be because children, when hospitalized, are more likely to seek care in technologically and clinically advanced facilities. However, it is unclear whether the IT adopted is calibrated for optimal pediatric use. More research on the use of IT is needed and should focus on other pediatric clinical settings as well. PMID- 17426614 TI - When should a multicampus hospital be considered a single entity for public reporting on patient safety issues? AB - OBJECTIVE: Attempts to provide information to consumers about patient safety on specific hospitals have conflicted with organization self-perceptions and led to confusion among the general public. This article presents organizational theory framework and criteria to classify organizations as single versus multiple reporting entities. PARTICIPANTS AND METHODS: Operational definitions are presented. A case study comparing institutions both within and across state boundaries in the Mayo Clinic Health System is used to demonstrate their utility. The study includes analysis of an employee survey on employee satisfaction and patient safety climate in 2004 among nurses and physicians at the 2 Mayo Clinic hospitals in Rochester, Minn. RESULTS AND CONCLUSIONS: The criteria for a single organization are more strongly supported for the Mayo Clinic hospitals located in the same city than for hospitals in the same system but separated geographically. Although there is debate about the measurement of organizational culture, employee surveys provide some evidence of a commonality across hospitals in the same city. The case study comparing institutions both within and across state boundaries in the Mayo Clinic Health System demonstrate the utility of the proposed criteria. PMID- 17426615 TI - The Patient Safety Leadership Academy at the University of Pennsylvania: the first cohort's learning experience. AB - BACKGROUND: We based the Patient Safety Leadership Academy (PSLA) on the premise that improving management skills could improve patient safety and employee satisfaction. STUDY DESIGN: Fellows completed baseline surveys on leadership skills knowledge, patient safety knowledge, and program goals. They completed the same surveys 7 months later at the final PSLA session. The fellows also completed a survey assessing how PSLA improved expertise and comparing PSLA to other patient safety learning opportunities. Matched pairs t tests were used to compare baseline and postprogram results. RESULTS: Baseline scores indicated appropriateness of focusing on leadership, with average leadership knowledge (2.48) significantly lower than patient safety knowledge (3.22). For patient safety, postprogram results were significant for 8 of 10 questions. All results were significant for leadership. Fellows also rated skills covered by the curriculum on a scale of 1 to 10. For all areas, the median score for knowledge gained was 7. When compared with other patient safety learning experiences, participants rated PSLA as 4 or 5, where 1 indicated the other experience much more valuable and 5 much more valuable. CONCLUSIONS: PSLA demonstrates that leadership skills are perceived as important by physicians and managers in surgical areas. This study demonstrated that a leadership skills approach to patient safety training could improve knowledge in specific leadership areas and general patient safety. PMID- 17426616 TI - Does implementation of evidence-based guidelines follow the available evidence? Implementation of hypertension guidelines at Veterans Affairs medical centers. AB - BACKGROUND: We sought to determine whether the proportion of strategies adopted in the implementation of evidence-based clinical guidelines by Department of Veterans Affairs medical centers (VAMCs) reflects an evidence-based assessment of the implementation strategies' relative effectiveness that was widely disseminated among VAMCs. METHODS: We used data from a multisite observational study we conducted to test a model of changing clinical behaviors. For that study, we had compiled information from key informants at 43 VAMCs about strategies for implementing the VA's new hypertension management guidelines. We had subsequently classified the reported strategies into 41 categories. For this study, we matched these 41 categories to the 8 broad categories of implementation strategies in terms of which effectiveness of strategies had been reported to VAMCs. We compared the 8 categories' rank order on effectiveness with their ranking by number of VAMCs adopting the strategies. RESULTS: The effectiveness based rank ordering does not correspond with rank order based on percentage of VAMCs adopting (Spearman correlation coefficient = -0.4). DISCUSSION: Results suggest that institutions, like practitioners, face barriers related to knowledge, attitudes, and behavior-including, in particular, insufficient resources-that impede their following the evidence when choosing guideline implementation strategies. PMID- 17426617 TI - QMHC interview. Integrating quality management efforts in a complex organization: a panel discussion with Nancy Lorenzi, Wright Pinson, MD, and John M. Starmer, MD. PMID- 17426620 TI - Cervical electromyogram profile differences between patients of neck pain and control. AB - STUDY DESIGN: A comparative analysis of electromyogram (EMG) signals of patients of cervical pain and normal controls. OBJECTIVES: To determine the differences between frequency and time domain parameters of EMG signals of patients of cervical pain and normal controls. SUMMARY OF BACKGROUND DATA: No diagnostic technique has emerged as a satisfactory tool for identification of spinal pain. METHOD: Seventeen male and 17 female chronic neck pain patients without cervical radiculopathy were recruited through neurology EMG clinic. The controls consisted of 30 male and 33 female subjects with no history of neck pain in the past 12 months. All subjects performed flexion, left anterolateral flexion, left lateral flexion, left posterolateral extension, and extension to pain threshold/20% maximum voluntary contraction and pain tolerance/maximum voluntary contraction in random order. The descriptive statistics for body weight normalized strength, normalized peak EMG, time to onset, time to peak, median frequency, mean power frequency, and frequency bands were calculated. These variables were subjected to analysis of variance and logistic regression to distinguish between patients and controls. RESULTS: The normalized peak EMG of patients was significantly greater than those of controls in both maximal and submaximal exertions (P < 0.01). Whereas there was no consistent pattern in time to peak EMG, the time to onset of EMG revealed that the left sternocleidomastoid was always recruited before the onset of torque. A lack of significant difference in the median frequency of the 2 samples indicates that the pain did not disturb the muscle conduction velocity. Using discriminant logistic regression on frequency domain and time domain parameters, up to 97% of patients and controls were correctly classified with the resubstitution method. CONCLUSION: Surface EMG can be used successfully in distinguishing chronic pain patients and controls, and efficacy of treatment regimes. PMID- 17426621 TI - Cervical range of motion and cephalic kinesthesis: ultrasonographic analysis by age and sex. AB - STUDY DESIGN: Exploratory experimental design. OBJECTIVE: To assess cervical range of motion (ROM) and ability to return the head to a neutral position in healthy subjects according to age and sex. SUMMARY OF BACKGROUND DATA: Cervical ROM and ability to return the head to a neutral position have been studied, but no data on the influence of age and sex in this movement is available. METHODS: We divided 232 healthy volunteers able to sit for 20 minutes into 6 age groups: 15-24, 25-34, 35-44, 45-54, 55-65, and older than 65 years. Zebris 3-dimensional ultrasonography measured the active cervical ROM and ability to return the head to the neutral position. Bone landmarks and vertical position were defined for each subject. Recorded data were neutral position, lateral bending (right and left), flexion-extension, and axial rotation (right and left). We evaluated patients' ability, without external intervention, to return the head to a self defined neutral position. RESULTS: ROM was affected by age as assessed by analysis of variance (F = 27.8 in the sagittal plane, F = 12.1 in the frontal plane, and F = 19.7 in the axial plane; all P < 0.0001), but neither sex nor age affected ability to return the head to a neutral position (F = 0.615, P = 0.688 on the left; F = 0.808, P = 0.545 on the right). Maximal ROM was observed for right axial rotation. CONCLUSIONS: We provide a database by age and sex for patient ability to return the head to the neutral position and for cervical ROM. This database could be used as a reference in clinical applications. PMID- 17426622 TI - Surgical outcome of drop foot caused by degenerative lumbar diseases. AB - STUDY DESIGN: A total of 46 patients undergoing lumbar spine surgery for degenerative lumbar disease and presenting with drop foot were included in this retrospective study. OBJECTIVE: To determine which preoperative patients' symptoms were of statistical significance in their effect on surgical outcome. SUMMARY OF BACKGROUND DATA: Drop foot is a neuromuscular condition that results in palsy of the ankle dorsiflexion and is a major problem in Japanese daily life. Few studies have described the effect of a surgical intervention on drop foot associated with degenerative lumbar disorders and the factors that affect the surgical outcome. The manual muscle test can be used to determine the muscular strength of the tibialis anterior. Drop foot is then defined as a score below 3 out of 5. METHODS: Patient medical history, preoperative tibialis anterior strength, presence of leg pain, and duration of palsy were recorded and compared with surgical outcome. RESULTS: Of patients, 61% recovered from drop foot after surgery. Patients with a tibialis anterior score of 2-3- and those suffering from palsy for a shorter duration of time showed better surgical results. Cases without leg pain were also shown to be treated effectively with surgery. CONCLUSIONS: Palsy duration and preoperative strength were factors that most affected drop foot recovery following surgical intervention for spinal degeneration. PMID- 17426623 TI - Upper-airway obstruction after short posterior occipitocervical fusion in a flexed position. AB - STUDY DESIGN: Case report. OBJECTIVE: To stress the importance of the fusion angle of the occipitocervical spine based on an unusual case of upper-airway obstruction after a posterior fusion from the occipital bone to the second cervical vertebra (O-C2) in a flexed position. SUMMARY OF BACKGROUND DATA: It is well known that cervical malalignment after occipito-cervicothoracic fusion may cause dysphagia or, rarely, dyspnea. However, to the best of our knowledge, there have been no previous English reports of prolonged upper-airway obstruction after an O-C2 fusion. METHODS: We present the case of a 77-year-old woman with rheumatoid arthritis, who developed an upper-airway obstruction immediately after an O-C2 fusion. She was reintubated immediately and extubated the next day. She again suffocated suddenly 3 days after surgery, and a tracheotomy was performed. Suspecting that the main cause of the airway obstruction was not only pharyngeal edema, but also the fixture of the upper cervical angle in a flexed position, we changed the angle to the neutral position 14 days after surgery. RESULTS: After revision surgery, the upper-airway obstruction disappeared. CONCLUSION: An adequate fixation angle is necessary to avoid airway obstruction after an occipitocervical fusion, even for short upper cervical fusions, especially in patients with rheumatoid arthritis. PMID- 17426624 TI - Halo pin insertion-associated brain abscess: case report and review of literature. AB - STUDY DESIGN: A case report of a patient with a halo pin-associated brain abscess and a review of literature. OBJECTIVES: To report a rare complication of halo pin insertion-associated brain abscess, and to discuss the diagnostic and treatment approach to its management. SUMMARY OF BACKGROUND DATA: Halo orthosis is a commonly used, well-tolerated spinal stabilizing device. However, on rare occasions, it can penetrate the inner table of the cranium, resulting in abscess formation that needs to be investigated urgently and treated promptly. METHODS: A 23-year-old male sustained a C2 vertebral fracture from a motor vehicular accident and was placed in halo traction. Two and a half months later, he noted loosening of the right occipital halo pin, which was tightened in the clinic. Fourteen days after pin tightening, he developed right-sided headaches, and it was decided to remove the halo traction. After the halo removal, the patient noted purulent discharge from the right occipital pin site, worsening headache, and associated nausea and vomiting. Magnetic resonance imaging of the brain demonstrated a right parietal lobe abscess and a sinus tract extending through the overlying calvarium. The patient underwent an evacuation of the abscess, and the culture was positive for methicillin-resistant Staphylococcus aureus. The patient was treated with intravenous vancomycin for 6 weeks. RESULT: The patient reported mediated improvement after surgery and on a follow-up visit was doing well without any neurologic sequelae. CONCLUSION: Halo pin-associated brain abscess is a rare but extremely important complication requiring prompt diagnosis and immediate intervention. Halo pin loosening with signs of local infection, fever, headaches, or seizures needs to be investigated urgently with neuroimaging. We conclude that with early diagnosis and appropriate treatment, serious morbidity and mortality can be avoided. PMID- 17426626 TI - Preliminary evaluation of a computer-assisted tool for the design and adjustment of braces in idiopathic scoliosis: a prospective and randomized study. AB - STUDY DESIGN: Prospective and randomized clinical study. OBJECTIVES: To evaluate the correction of the spine obtained using a 3-dimensional visualization software tool developed to assist the design and adjustment of braces compared with the correction obtained with the conventional method in a cohort of subjects with adolescent idiopathic scoliosis (AIS). SUMMARY OF BACKGROUND DATA: The optimal design and adjustment of trim lines, pad placement, and areas of relief for the Boston brace system in AIS are currently done using clinical examination and coronal radiographs. Correction of spinal curves in the coronal plane has been achieved with this technique, but 3-dimensional correction has yet to be demonstrated. METHODS: Forty-eight consecutive subjects with AIS requiring treatment with a Boston brace were prospectively entered in the study. For 24 patients (test group), brace design and adjustment was obtained using the computer-assisted tool combining surface topography, surface pressure measurement, and 3-dimensional reconstructions of the trunk, while design and adjustment for the remaining subjects (control group) was done in the conventional manner. Immediate in-brace correction of the spine at the initial visit was compared in both groups. RESULTS: Both groups were comparable in terms of age, sex, curve type, and average deformity in both the coronal and sagittal planes. The average prebrace thoracic deformity was 35 degrees of Cobb angle, while the average lumbar curve was 32 degrees in the test group and 35 degrees in controls. A statistically and clinically significant improvement in correction of coronal curves and of curves in the plane of maximal deformity was found for both thoracic and lumbar curves in both groups, but the improvement was significantly greater in the test group. The average in-brace correction in the test group was 12 degrees +/- 7 degrees compared with 7 degrees +/- 5 degrees in the control group for thoracic curves, while the average in-brace correction in the test group was 10 degrees +/- 5 degrees compared with 6 degrees +/- 5 degrees in the control group for lumbar curves. Similar average corrections were detected in the plane of maximal deformity. In addition, a significant improvement in the orientation of the plane of maximum deformity from 37 degrees to 23 degrees for lumbar curves was noted only in the test group, indicating that true 3 dimensional correction by the brace was obtained in this group. CONCLUSION: It is possible to improve the design and adjustment of braces in AIS and to achieve 3 dimensional correction of scoliotic curves with the use of a computer-assisted tool allowing 3-dimensional visualization of the spinal curves and the external shape of the trunk. PMID- 17426627 TI - C1 anatomy and dimensions relative to lateral mass screw placement. AB - STUDY DESIGN: An anatomic surface osteometric analysis of cadaveric cervical spines. OBJECTIVE: To assess the feasibility of placing a 3.5-mm cortical screw in the lateral mass of C1 in a large number of specimens for the purpose of gaining internal fixation for various conditions. SUMMARY OF BACKGROUND DATA: Previous studies have addressed surface dimensions of the atlas vertebra and computerized tomography-measured dimensions of the lateral masses of the atlas. These studies used a limited number of specimens with potentially homogeneous origins. METHODS: A total of 120 atlas (C1) vertebrae for a total of 240 lateral masses were examined and external measurements obtained to assess the feasibility of placing a 3.5-mm cortical screw in the lateral mass. RESULTS: The minimum lateral mass dimensions found from 240 C1 lateral masses were 13.15 mm anterior posterior, 4.22 mm medial-lateral, and 4.73 mm cephalocaudal. The height of the posterior arch at the groove for the vertebral artery (pedicle analog) was less than 4 mm in 46 of 240 (19.2%) arches. CONCLUSIONS: It is feasible to safely and reproducibly place a 3.5-mm cortical screw in the lateral mass of C1 when the appropriate starting point and trajectory of the screw are chosen. PMID- 17426628 TI - Age-related changes in expression of tissue inhibitor of metalloproteinases-3 associated with transition from the notochordal nucleus pulposus to the fibrocartilaginous nucleus pulposus in rabbit intervertebral disc. AB - STUDY DESIGN: Experimental study on age-related changes in expression of tissue inhibitor of metalloproteinases-3 (TIMP-3) associated with transition from notochordal nucleus pulposus (NP) to fibrocartilaginous NP in rabbit intervertebral disc (IVD). OBJECTIVES: To identify roles of notochordal NP in extracellular matrix (ECM) metabolism of IVD. SUMMARY OF BACKGROUND DATA: One of most interesting properties of TIMP-3 is to inhibit aggrecanases in addition to matrix metalloproteinases. Balance of aggrecanase/TIMP-3 is critical to maintain homeostasis of ECM metabolism. METHODS: Four-week-old and 160-week-old male Japanese white rabbits were used. Age-related changes in IVDs were evaluated histologically using previously established grading system. Immunohistochemistry of TIMP-3 and semiquantitative reverse transcriptase-polymerase reaction (RT-PCR) of TIMP-3, a disintegrin and metalloproteinases with thrombospondin motifs (ADAMTS) 4, 5, and transforming growth factor-beta1 (TGF-beta1), were conducted. RESULTS: Semiquantitative assessment of histologic changes indicated that 4-week old rabbit was equivalent to fetus to 2-year-old human and 160-week-old rabbit was equivalent to 11- to 30-year-old human, particularly 11- to 16-year-old, which corresponds to transition period from notochordal to fibrocartilaginous NP. Immunohistochemistry revealed that TIMP-3 was positive in 4-week-old rabbit only. Semiquantitative RT-PCR revealed that levels of expressions of TGF-beta1 and TIMP 3 mRNAs in 4-week-old were significantly higher than those in 160-week-old rabbits. There was no significant difference in expression of ADAMTS4 mRNA. ADAMTS5 mRNA was not detected or extremely low in both groups. Expression of TIMP 3 mRNA in NP was upregulated by TGF-beta1 but was not affected by IL-1beta. On the contrary, expression of ADAMTS4 mRNA was not upregulated by TGF-beta1 but was upregulated by IL-1beta. CONCLUSIONS: Levels of expression of TIMP-3 in notochordal NP were significantly lower in 160-week-old rabbits than those in 4 week-old rabbits. Decrease in expression of TIMP-3, possibly mediated in part by TGF-beta1, may cause imbalance of ADAMTS4/TIMP-3 ratio at transition period from notochordal to fibrocartilaginous NP. PMID- 17426629 TI - Axonal growth potential of lumbar dorsal root ganglion neurons in an organ culture system: response of nerve growth factor-sensitive neurons to neuronal injury and an inflammatory cytokine. AB - STUDY DESIGN: The axonal growth potential of dorsal root ganglion (DRG) neurons in an organ culture system was investigated. OBJECTIVE: To examine the effects of neuronal injury and tumor necrosis factor-alpha (TNF-alpha) on the axonal growth potential of 2 types of nociceptive DRG neurons: nerve growth factor (NGF) sensitive and glial cell line-derived neurotrophic factor (GDNF)-sensitive neurons. SUMMARY OF BACKGROUND DATA: Nerve ingrowth into the disc is recognized to be one of the causes of discogenic pain. Almost all of these disc-innervating neurons are NGF-sensitive. The axonal growth potential of NGF-sensitive neurons has not been investigated. METHODS: Adult Sprague-Dawley rats were used for immunohistochemistry (n = 7) and cell viability studies (n = 6). Bilateral L3-L5 DRGs, which were successfully removed without damage, were noncultured or cultured in serum-free medium containing TNF-alpha at 0, 0.01, 0.1, and 1 ng/mL for 48 hours (n = 5, each treatment). The DRGs were then immunostained for activating transcription factor 3 (ATF3, a marker for injured neurons) or double stained for growth-associated protein 43 (GAP-43, a marker for axonal growth) with calcitonin gene-related peptide (CGRP, a marker for NGF-sensitive neurons) or isolectin B4 (IB4, a marker for GDNF-sensitive neurons). Cell viability was assessed by a lactate dehydrogenase (LDH) assay and an MTS assay (n = 6, each treatment). RESULTS: Immunoreactive evidence of injured neurons (ATF3 positive) was frequently observed in cultured DRGs, but never in noncultured DRGs. The percentage of neurons exhibiting axonal growth potential (GAP-43 immunoreactive) was significantly higher for NGF-sensitive neurons than for GDNF-sensitive neurons at any concentration of TNF-alpha. More than 95% of the cultured neurons were viable. CONCLUSIONS: The results suggest that the cultured DRG neurons exhibit pathologic changes similar to those found in injured neurons. NGF sensitive neurons, which include disc-innervating neurons, may have a greater potential to extend their axons in response to neuronal injury under pathologic conditions in the presence of TNF-alpha than GDNF-sensitive neurons. PMID- 17426630 TI - A new in vivo animal model to create intervertebral disc degeneration characterized by MRI, radiography, CT/discogram, biochemistry, and histology. AB - STUDY DESIGN: A new in vivo sheep model was developed that produced disc degeneration through the injection of 5-bromodeoxyuridine (BrdU) into the intervertebral disc. This process was studied using magnetic resonance imaging (MRI), radiography, CT/discogram, histology, and biochemistry. OBJECTIVES: To develop a sheep model of intervertebral disc degeneration that more faithfully mimics the pathologic hallmarks of human intervertebral disc degeneration. SUMMARY OF BACKGROUND DATA: Recent studies have shown age-related alterations in proteoglycan structure and organization in human intervertebral discs. An animal model that involves the use of age-related changes in disc cells can be beneficial over other more invasive degenerative models that involves directly damaging the matrix of disc tissue. METHODS: Twelve sheep were injected with BrdU or vehicle (phosphate-buffered saline) into the central region of separate lumbar discs. Intact discs were used as controls. At the 2-, 6-, 10-, and 14-week time points, discs underwent MRI, radiography, histology, and biochemical analyses. A CT/discogram study was performed at the 14-week time point. RESULTS: MRI demonstrated a progressive loss of T2-weighted signal intensity at BrdU-injected discs over the 14-week study period. Radiograph findings included osteophyte and disc space narrowing formed by 10 weeks post-BrdU treatment. CT discography demonstrated internal disc disruption in several BrdU-treated discs at the 14 week time point. Histology showed a progressive loss of the normal architecture and cell density of discs from the 2-week time point to the 14-week time point. A progressive loss of cell proliferation capacity, water content, and proteoglycans was also documented. CONCLUSIONS: BrdU injection into the central region of sheep discs resulted in degeneration of intervertebral discs. This progressive, degenerative process was confirmed using MRI, histology, and by observing changes in biochemistry. Degeneration occurred in a manner that was similar to that observed in human disc degeneration. PMID- 17426631 TI - Effect of immunomodulation with human interferon-beta on early functional recovery from experimental spinal cord injury. AB - STUDY DESIGN: Electron and light microscopic changes, neutrophil infiltration, and lipid peroxidation in the spinal cord and early neurologic examination were studied in rats. OBJECTIVE: To examine the effects of immunomodulator treatment with recombinant human interferon-beta after spinal cord contusion injury. SUMMARY OF BACKGROUND DATA: Immunomodulator treatment with interferon-beta has been the subject of extensive studies, but mainly in relation to multiple sclerosis. Recently, it was reported that interferon-beta possessed significant neuroprotection after experimental transient ischemic stroke. However, to our knowledge, there have been no previous reports about the neuroprotective effect of interferon-beta after spinal cord injury. METHODS: Rats were randomly allocated into 5 groups. Group 1 was control and after clinical examination, normal spinal cord samples were obtained. Group 2 was introduced 50 g/cm contusion injury. Group 3 was vehicle, immediately after trauma 1 mL of physiologic saline was injected. Group 4 was given 30 mg/kg methylprednisolone sodium succinate intraperitoneally immediately after trauma. Group 5 was given 1 x 10(7) IU interferon-beta immediately and 0.5 x 10(7) IU interferon-beta 4 hours after trauma. Animals were examined by inclined plane and Basso-Beattie-Bresnahan scale 24 hours after trauma. Spinal cord samples obtained following clinical evaluations. Neutrophil infiltration was evaluated by myeloperoxidase activity and lipid peroxidation was estimated by thiobarbituric acid test. Electron and light microscopic results were also performed to determine the effects of interferon-beta on tissue structure. RESULTS: Interferon-beta treatment improved neurologic outcome, which was supported by decreased myeloperoxidase activity and lipid peroxidation. Electron and light microscopic results also showed preservation of tissue structure in the treatment group. CONCLUSIONS: Immunomodulator treatment with interferon-beta possesses obvious neuroprotection after acute contusion injury to the rat spinal cord. PMID- 17426632 TI - Torsion-induced pressure distribution changes in human intervertebral discs: an in vitro study. AB - STUDY DESIGN: Biomechanical testing of human cadaveric lumbar specimens was performed to evaluate the effects of torsional torque on intradiscal pressure and disc height. OBJECTIVE: Evaluate the effects of small torsion torques on intradiscal pressure and disc height in human lumbar specimens. SUMMARY OF BACKGROUND DATA: Nuclear depressurization in addition to an instantaneous disc height increase were found in previous porcine research when small (<2 degrees) axial vertebral rotations were applied. If applicable to human spines, this phenomenon may support spinal manipulation for the relief of low back pain. METHODS: Six human lumbar cadaveric functional spine units (FSU) were loaded in the neutral position with 600 N axial compression. Intranuclear pressure measurements were then obtained at 0, 0.5, 1.0, and 2.0 Nm of torsion. Posterior elements were removed and measurements were repeated for the disc body unit (DBU). RESULTS: There was no statistically significant difference in nuclear pressure or intervertebral disc height with different torsion torques among or between the FSUs and DBUs. However, a disc height increase ranging from 0.13 mm to 0.16 mm occurred with the insertion of a 1.85-mm diameter pressure probe cannula. CONCLUSIONS: Small torsion torques showed no significant difference in intradiscal pressures or disc heights. This is an unlikely mechanism for the perceived benefits of spinal manipulation. PMID- 17426633 TI - A radiostereometric analysis of the Bryan Cervical Disc prosthesis. AB - STUDY DESIGN: Prospective, radiostereometric study of the Bryan Cervical Disc prosthesis (Medtronic Sofamor Danek, Memphis, TN) for the treatment of a single level disc disease of the cervical spine. OBJECTIVE: To study the stability of the titanium endplates of the disc prosthesis at the interface between the bone and prosthesis. SUMMARY OF BACKGROUND DATA: Cervical disc prosthesis is a motion sparing technology in which the longevity is dependent on initial fixation as well as secondary fixation with bone ingrowth into the surface of the device. Little is known regarding the stability of this fixation with the currently studied prosthesis. METHODS: Eleven patients with symptomatic cervical radiculopathy underwent implantation of a radiostereometry modified Bryan prosthesis after a standard anterior cervical discectomy. The adjacent vertebrae were perioperatively marked with tantalum markers. The patients were then frequently studied with radiostereometric radiographs and evaluated for pain, and neurologic and physical function for 2 years. RESULTS: Both titanium endplates of the device were immediately stable in 5 patients. Small but measurable changes (micromotions) were recorded in 4 patients until the 3-month examination and in 2 patients at the 3-6-month time period. All prostheses continued to be stable after 6 months until the final assessment at 2 years after surgery. The recorded micromotions did not influence the clinical results. CONCLUSIONS: The Bryan prosthesis is immediately stable in many patients and is securely fixed to the bone within 3-6 months in all patients. The result of this study suggests that there is sufficient bone ingrowth on the coated surface of the Bryan prosthesis endplates to stabilize securely the prosthesis. PMID- 17426635 TI - Diagnostic accuracy and reliability of fine-cut CT scans with reconstructions to determine the status of an instrumented posterolateral fusion with surgical exploration as reference standard. AB - STUDY DESIGN: Accuracy of a diagnostic test referenced to the gold standard. OBJECTIVES: This study evaluated the reliability and accuracy of fine-cut computed tomography scans with coronal and sagittal reconstructions to determine the status of an instrumented posterolateral fusion by using surgical exploration as the reference standard. SUMMARY OF BACKGROUND DATA: There is still a need for a reliable and accurate noninvasive method to determine the status of a spinal fusion. METHODS: Three spine surgeons reviewed 93 prerevision fine-cut CT scans over 163 fused levels of consecutive patients who had revision surgery after an instrumented posterolateral lumbar fusion. The facet joints and posterolateral gutters at each level were classified as fused or not. The surgeons were unaware of the findings on surgical exploration. Interobserver variability and likelihood ratios for a solid fusion when both, one, or none of the facets and when both, one, or none of the posterolateral gutters were fused, were calculated. RESULTS: There were 42 males and 51 females with a mean age of 57 years (range, 19-86 years) at revision. On exploration, there were 32 (19.6%) nonunions over 163 levels. The kappa for interobserver variability for evaluating facet fusions (0.42) was moderate and for posterolateral fusions (0.62) was substantial. The probability of a solid fusion on exploration was higher when both posterolateral gutters were fused on CT scan (89%) than when both facets were fused on CT scan (74%). When both facets and both posterolateral gutters were fused on CT scan, the probability of a solid fusion on exploration is 96%. The absence of fusion of one or both facets or one posterolateral gutter were poor predictors of nonunion on surgical exploration. CONCLUSIONS: The CT scan reading of either one or both posterolateral gutters fused or both facets fused were moderately predictive of a solid fusion on surgical exploration. Fine-cut CT scans with reconstructions are moderately predictive of the presence of nonunion when both facets are not fused. PMID- 17426636 TI - Magnetic resonance imaging and magnetic resonance myelography in the presurgical diagnosis of lumbar foraminal stenosis. AB - STUDY DESIGN: Retrospective case series with a control group. OBJECTIVE: To measure the diagnostic performance of magnetic resonance imaging (MRI) and MR myelography (MRM) for symptomatic foraminal stenosis in patients who need surgery. SUMMARY OF BACKGROUND DATA: MR images are extensively used in the evaluation of foraminal stenosis and are often used to evaluate nerves exiting from the foramen. There has been no published report of the diagnostic performance of these imaging methods (MRI and MRM). METHODS: Diagnostic performances were studied in 90 patients in whom the site of the stenosis was confirmed by means of selective decompression surgeries. The disease prevalence among patients was 26% (23 of 90 patients). The disease prevalence among foramens was 3% (25 of 936 foramens). The prevalence of abnormal findings in 27 asymptomatic volunteers was also studied. Two blinded observers interpreted foraminal narrowing on combinations of sagittal and axial MR images, abnormalities of the course of the nerve root in the foramen, and spinal nerve swelling on MRM. RESULTS: The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MRI for the diagnosis of symptomatic foraminal stenosis were 96%, 67%, 4%, and 100%, respectively. The corresponding values for abnormal nerve root course on MRM were 96%, 83%, 7%, and 100%, respectively, and for spinal nerve swelling on MRM were 60%, 99%, 35%, and 99%, respectively. CONCLUSIONS: Compared with conventional MRI, MRM affords more specific information for the presurgical diagnosis of symptomatic foraminal stenosis. PMID- 17426637 TI - Abnormal rib number in childhood malignancy: implications for the scoliosis surgeon. AB - STUDY DESIGN: Retrospective review. OBJECTIVE: To determine if rib anomalies are present in pediatric malignancies in the United States. SUMMARY OF BACKGROUND DATA: Scoliosis surgeons view radiographs of the entire spine, counting the number of ribs. A European study noted that rib anomalies were more common in certain malignancies. We wished to determine if this is also true in the United States. If so, the potential for screening, early detection of malignancy, and a better understanding of tumor biology is possible. METHODS: A retrospective review of 218 children with malignancy and a control group of 200 children with polytrauma or suspected child abuse was performed. Chest radiographs were reviewed to determine the number of ribs, and the presence of rib anomalies. 24 ribs was considered normal, <24 or >24 was considered abnormal. P < 0.05 was considered significant. RESULTS: The average age was 6.8 +/- 5.5 years and number of ribs was 23.8 +/- 0.6. Rib number was normal in 86.8%. There were significant differences between the malignancy and control groups in age (control, 5.7 +/- 5.1 years; malignancy, 7.8 +/- 5.7 years, P = 0.00007), rib number (control, 23.9 +/- 0.5; malignancy, 23.7 +/- 0.7, P = 0.001), and normal/abnormal rib counts (control, 92% normal; malignancy, 82% normal, P = 0.003). In the malignant group, 50% had a lymphoproliferative malignancy, 33% a solid tumor, and 17.0% a neural tumor. Neural malignancies had a higher incidence of rib abnormalities compared with lymphoproliferative or solid malignancies (P = 0.01). Relative to the control group, those with a neural and lymphoproliferative malignancy were 6.23 (95% CI, 2.7-14.5) and 2.0 (95% CI, 1.0-4.1) times more likely to have an abnormal rib count. CONCLUSIONS: Homeobox genes, important in vertebral and rib sequencing, are abnormally expressed in many different malignancies. This association is a question of great interest. What is the potential for rib number being used as a predictor of childhood malignancy? Can these findings be expanded to adults? These questions require further research. The association noted in this study is interesting but should not yet be used to alarm parents regarding an increased risk of malignancy in their children. PMID- 17426638 TI - Intraoperative monitoring of motor-evoked potentials in children undergoing spinal surgery. AB - STUDY DESIGN: Clinical case series. OBJECTIVE: To study the combined use of modifications of stimulation methods and adjustments of anesthetic regimens on the reliability of motor-evoked potential (MEP) monitoring in a large group of children undergoing spinal surgery. SUMMARY OF BACKGROUND DATA: Monitoring of MEPs is advocated during spinal surgery, but systematic data from children are sparse. METHODS: A total of 134 consecutive procedures in 108 children <18 years of age were analyzed. MEPs were elicited by transcranial electrical stimulation (TES) and supplemented by temporal and spatial facilitation. The standard anesthesia regimen consisted of propofol, nitrous oxide, and remifentanil. Propofol was replaced with ketamine if no reliable MEPs could be recorded. In children <6 years of age, a ketamine-based anesthesia was used. RESULTS: With temporal facilitation alone, reliable MEPs were obtained in 78% (105 of 134) of the procedures and, if combined with spatial facilitation, in 96% (129 of 134) of the procedures. Reliable MEPs were documented in 98% (111 of 113) of children >6 years and in 86% (18 of 21) in children <6 years of age. CONCLUSIONS: Combining spatial facilitation with a TES protocol improved monitoring of corticospinal motor pathways during spinal surgery in children. A ketamine-based anesthetic technique was preferred in children <6 years of age. PMID- 17426639 TI - Bacterial shedding in common spine surgical procedures: headlamp/loupes and the operative microscope. AB - STUDY DESIGN: Simulated discectomy was performed using 1) headlamp/loupes, 2) the operative microscope, or 3) neither; and bacterial shedding was assessed using air sampler/settle plate techniques. OBJECTIVES: To determine the potential contribution of these 2 commonly used systems of magnification/illumination to postoperative infection by using a controlled, experimental setting to limit confounding factors. SUMMARY OF BACKGROUND DATA: Postoperative infections following surgery on the degenerative lumbar spine generally range from 0.6% to 6%. Headlamp/loupes and the operative microscope may contribute to potential infection, but, to date, the contribution to shedding rates while using these tools has not been assessed. METHODS: Seventy samples from simulated discectomies within the usual sterile operative setting were collected using a 2-stage air sampler/settle plate technique. Group 1 (30 samples) used the operative microscope, Group 2 (30 samples) used headlamp/loupes, and Group 3 (10 samples) used neither as a control. Collected samples were examined for bacterial growth using blood agar plates and were assessed qualitatively and quantitatively. RESULTS: Greater than half of the collected samples in Groups 1 and 2 demonstrated bacterial growth with coagulase negative Staphylococcus being found in nearly all positive samples and secondary species being common. No statistical differences between the 2 groups were noted for number of samples with growth, species of bacterial growth, number with growth of secondary species, or colony counts. Both groups demonstrated significantly greater number of samples with growth relative to the control Group 3. CONCLUSIONS: The use of headlamp/loupes or the operative microscope is associated with bacterial shedding. Proper techniques of cleaning, storage, and draping should be used to minimize their contribution to potential postoperative infection. No significant difference was noted between the 2 test groups, suggesting that infection risk should not come into play when choosing techniques of illumination/magnification. PMID- 17426640 TI - The reliability of the clinical tests and questions recommended in international guidelines for low back pain. AB - STUDY DESIGN: An intertester reliability study of the questions and tests recommended in guidelines for the management of low back pain (LBP). OBJECTIVE: This study undertook a reliability study to evaluate the reliability of the items of the LBP clinical examination with a large sample of LBP patients. SUMMARY OF BACKGROUND DATA: A crucial part of the diagnostic triage process, recommended by many national and international guidelines for the management of LBP, is the clinical examination. The questions and tests used in this process have never been rigorously evaluated for their intertester reliability in first contact clinicians who are not medically trained. METHODS: Patients, referred to physiotherapy departments across the United Kingdom with LBP (n = 301) were recruited in a sample of convenience. The 50 questions and physical tests were administered by a physiotherapist and then repeated by another physiotherapist within the same day. Data were analyzed using kappa and weighted kappa correlation coefficients (kappa). Confidence intervals (95% CIs) were calculated. RESULTS: Eighty-six percent (n = 43) of the questions and test demonstrated kappa of 0.41 (fair agreement) or above. Five questions and 2 physical tests (prone knee bend and myotomal assessment) demonstrated agreement of only slight levels. CIs were generally narrow and the uncertainty regarding the kappa coefficients demonstrated correspondingly low. CONCLUSIONS: This study has rigorously evaluated the intertester reliability of the clinical examination process of the diagnostic triage. These clinicians generally demonstrated fair agreement when testing features of the "nerve root," "yellow" and "red flag" presentations recommended in international guidelines for the management of LBP and nonspecific LBP. However, reliance on single tests with only fair levels of agreement may be unwise. Further work is required to investigate the validity of the tests. PMID- 17426641 TI - Adolescent idiopathic scoliosis in twins: a population-based survey. AB - STUDY DESIGN: A questionnaire-based identification of adolescent idiopathic scoliosis (AIS) patients in a twin cohort. OBJECTIVE: The purpose of this study was to establish a scoliosis twin cohort to provide data on the heritability of AIS. SUMMARY OF BACKGROUND DATA: The etiology of AIS is still unclear, and the true mode of inheritance has yet to be established. Concordance rates in monozygotic twins have been reported to be between 0.73 and 0.92, and in dizygotic twins between 0.36 and 0.63. Studies on concordance in twin pairs provide a basis for analyzing the influence of genetic versus environmental factors. METHODS: All 46,418 twins registered in the Danish Twin Registry born from 1931 to 1982 were sent a questionnaire, which included questions about scoliosis. A total of 34,944 (75.3%) representing 23,204 pairs returned the questionnaire. RESULTS: A subgroup of 220 subjects considered to have AIS was identified, thus giving a prevalence of 1.05%. The concordant twin pairs were all monozygotic. Pairwise, the concordance rate was 0.13 for monozygotic and zero for dizygotic twin pairs; proband-wise concordance was 0.25 for monozygotic and zero for dizygotic pairs. The concordance of monozygotic and dizygotic pairs was significantly different (P < 0.05). CONCLUSION: We have found evidence for a genetic etiology in AIS, but the risk of developing scoliosis in 1 twin whose other twin has scoliosis is smaller than believed up until now. PMID- 17426642 TI - Epidemiological and physiological approaches to understanding the etiology of pediatric obesity: finding the needle in the haystack. AB - Recent increases in the prevalence of childhood obesity have created an urgent need for preventive strategies, but such strategies in turn depend on an improved understanding of the etiology of pediatric obesity. There is a dearth of evidence of the cause of pediatric obesity at present, with much of the literature of limited quality, inconclusive, and contradictory. The present review highlights the paradox of energy imbalance-its apparent simplicity but actual complexity-and the difficulties in etiologic research that arise from this complexity. The review identifies a number of emerging problems for etiologic studies. The review also makes a number of proposals that might improve future etiologic studies and provides a framework for integrating the diverse body of evidence of etiology that will become available in future. Gathering improved evidence of etiology, and then integrating and interpreting it, will take many years. In the meantime, an emphasis on developing more effective preventive interventions is necessary. PMID- 17426643 TI - Hypoxic preconditioning reverses protection after neonatal hypoxia-ischemia in glutathione peroxidase transgenic murine brain. AB - The effect of hypoxic preconditioning (PC) on hypoxic-ischemic (HI) injury was explored in glutathione peroxidase (GPx)-overexpressing mice (human GPx transgenic [hGPx-tg]) mice. Six-day-old hGPx-tg mice and wild-type (Wt) littermates were pre-conditioned with hypoxia for 30 min and subjected to the Vannucci procedure of HI 24 h after the PC stimulus. Histopathological injury was determined 5 d later (P12). Additional animals were killed 2 h or 24 h after HI and ipsilateral cerebral cortices assayed for GPx activity, glutathione (GSH), and hydrogen peroxide (H2O2). In line with previous studies, hypoxic PC reduced injury in the Wt brain. Preconditioned Wt brain had increased GPx activity, but reduced GSH, relative to naive 24 h after HI. Hypoxic PC did not reduce injury to hGPx-tg brain and even reversed the protection previously reported in the hGPx tg. GPx activity and GSH in hGPx-tg cortices did not change. Without PC, hGPx-tg cortex had less H2O2 accumulation than Wt at both 2 h and 24 h. With PC, H2O2 remained low in hGPx-tg compared with Wt at 2 h, but at 24 h, there was no longer a difference between hGPx-tg and Wt cortices. Accumulation of H2O2 may be a mediator of injury, but may also induce protective mechanisms. PMID- 17426644 TI - Retinoids increase lung elastin expression but fail to alter morphology or angiogenesis genes in premature ventilated baboons. AB - Retinoids regulate elastin synthesis by alveolar myofibroblasts and affect angiogenesis pathways, both of which are processes critical for alveolar development. Retinoids accelerate alveolarization in rodents and are now used therapeutically in premature infants at risk of bronchopulmonary dysplasia (BPD). This study examined the effects of retinoid supplementation on alveolar elastin expression and deposition and angiogenesis-related signaling in a primate model of BPD. Premature baboons delivered at 125 d of gestation after maternal steroid treatment were given surfactant and ventilated with minimal supplemental oxygen for 14 d with (n = 5) and without (n = 5) supplemental vitamin A (5000 U/kg/d) and compared with 140-d unventilated controls. Ventilatory efficiency index (VEI) and oxygenation index (OI) were not statistically different between ventilated treatment groups. Expression of vascular endothelial growth factor A (VEGF-A), fms-related tyrosine kinase 1 (Flt-1), and tyrosine kinase with immunoglobulin like and EGF-like domains 1 (TIE-1) was repressed by premature delivery and mechanical ventilation and was not altered by retinoid supplementation. Retinoid supplementation did not enhance alveolar angiogenesis. Elastin expression was repressed by premature delivery and extended ventilation, and retinoid supplementation increased elastin expression specifically in alveolar myofibroblasts within alveolar walls. These results suggest that the small decrease in mortality among premature infants receiving retinoid supplementation may not be mediated through enhanced alveolar development. PMID- 17426645 TI - GJB2 and GJB6 mutations in children with congenital cytomegalovirus infection. AB - Congenital cytomegalovirus (CMV) infection is a leading cause of sensorineural hearing loss (SNHL) in children. Whether connexin mutations are factors in the development of CMV-related hearing loss has not been explored. We examined gap junction protein beta-2 (GJB2) and gap junction protein beta-6 (GJB6) mutations in 149 children with congenital CMV infection and 380 uninfected neonates. Mutations in GJB2 and GJB6 were assessed by nucleotide sequencing and polymerase chain reaction (PCR) methods, respectively. The study population was predominantly African American, and 4.3% of the subjects were carriers of a connexin 26 mutation. The overall frequency of GJB2 mutations was significantly higher in the group of children with CMV infection and hearing loss (21%) compared with those with CMV infection and normal hearing (3%, p = 0.017) and the group of uninfected newborns (3.9%, p = 0.016). Eight previously reported mutations (M34T, V27I, R127H, F83L, R143W, V37I, V84L, G160S), and four novel mutations (V167M, G4D, A40T, and R160Q) were detected. None of the study children had the 342-kb deletion (delGJB6-D13S1830) in GJB6, which suggests that this mutation does not play a role in hereditary deafness in the African American population. Although GJB2 mutations were detected in children with and without CMV-related hearing loss, those with hearing loss had a higher frequency of GJB2 mutations. PMID- 17426646 TI - Pediatric HIV-1-specific cytotoxic T-lymphocyte responses suggesting ongoing viral replication despite combination antiretroviral therapy. AB - Human immunodeficiency virus-1 (HIV-1)-specific cytotoxic T-lymphocyte (CTL) responses are common in infected adults and usually exhibit rapid decay after combination antiretroviral therapy (ART). CTLs develop later in the first year of life, and the fate of HIV-1-specific responses in perinatally infected children after ART is less well described. HIV-1-specific CTL responses were measured in 17 perinatally infected children and adolescents (ages 3-20 y) receiving combination ART. Seven had prolonged viral suppression (<400 copies/mL) for 2.5 5.3 y and 10 had persistent viremia (median, 77,550 copies/mL). HIV-1-specific CTL responses were tested by interferon (IFN)-gamma enzyme-linked immunospot (ELIS-pot) assays using 53 overlapping peptide pools spanning the entire HIV-1 proteome. HIV-1-specific CTL responses were detected in 14 of 17 individuals. Responses to one to four viral proteins were found in eight of 10 individuals with persistent viremia and six of seven with prolonged viral suppression. The magnitude and breadth of CTL responses were similar between groups. HIV-1 specific CTL responses were present in the majority of perinatally infected subjects, irrespective of viremia at evaluation. Because ART-treated infected adults usually have rapid decay of responses, these data suggest viral replication below the limits of detection is more persistent in combination ART treated perinatally infected pediatric subjects. The long-term clinical implications of these findings remain to be determined. PMID- 17426647 TI - White matter volume and anisotropy in preterm children: a pilot study of neurocognitive correlates. AB - The objectives of this study were to evaluate the differences in whole brain white matter (WM) volume and anisotropy between preterm and term children and to determine the relationships with cognitive outcome. Twenty-five low birth weight (BW), preterm, neurologically normal children between 8.8 and 11.5 y of age were recruited for volumetric and diffusion-tensor magnetic resonance imaging (DTI), together with 13 age-matched term control subjects. Subsequent intelligence quotient (IQ) testing was performed for 21 preterm children within 6 mo of imaging studies. We computed the mean volume and fractional anisotropy (FA) of the whole brain WM and compared the differences between the two groups. Mean WM volume and FA were significantly lower in the preterm group (p = 0.014 and p < 0.001, respectively). Multiple regression analysis found both WM volume and FA to be independent variables significantly affecting full scale IQ (FSIQ) (r2 = 0.407, p = 0.021 and r2 = 0.496, p = 0.005, respectively) after adjusting for BW, gestational age (GA), and gender. In the evaluation of the whole brain WM of preterm children, we found that both volume and FA remain reduced at late childhood with both parameters significantly affecting long-term cognitive outcome. PMID- 17426648 TI - (TA)n UGT 1A1 promoter polymorphism: a crucial factor in the pathophysiology of jaundice in G-6-PD deficient neonates. AB - Increased heme catabolism has been reported in glucose-6-phosphate dehydrogenase (G-6-PD)-normal neonates who were also homozygous for (TA)7/(TA)7 (UGT1A1*28) uridine diphosphoglucuronate-glucuronosyltransferase 1A1 (UGT) promoter polymorphism (Gilbert syndrome). As G-6-PD deficiency is associated with increased hemolysis, we hypothesized that in G-6-PD-deficient neonates who also have the (TA)7/(TA)7 UGT promoter genotype, steady-state hemolysis would be even further increased. Male G-6-PD-deficient neonates were sampled for plasma total bilirubin (PTB), blood carboxyhemoglobin corrected for inhaled carbon monoxide in ambient air (COHbc) (an index of heme catabolism), and UGT (TA)n promoter genotype determination and compared with previously published G-6-PD-normal neonates. Although COHbc values were higher in the G-6-PD-deficient than in the G 6-PD-normal cohorts (0.97 +/- 0.32% of total Hb (tHb) versus 0.76 +/- 0.19% of tHb, p < 0.001), PTB values were similar (9.2 +/- 3.4 mg/dL versus 8.9 +/- 3.0 mg/dL, respectively, p = 0.3). Within the G-6-PD-deficient group, although COHbc values were alike between the three UGT promoter genotypes, PTB was higher in the (TA)7/(TA)7 homozygotes (11.1 +/- 4.0 mg/dL) compared with (TA)6/(TA)7 heterozygotes (9.1 +/- 3.2 mg/dL, p = 0.03) and wild-type (TA)6/(TA)6 homozygotes (8.8 +/- 3.4 mg/dL, p = 0.02). In the steady state, similar rates of hemolysis, but increased PTB in the G-6-PD- deficient, (TA)7/(TA)7 homozygotes, imply that (TA)7/(TA)7, homozygosity is central to increased PTB. PMID- 17426649 TI - Microvascular development in porcine right and left ventricular walls. AB - Patients with congenital heart disease who have a morphological right ventricle (RV) serving as a systemic ventricle have an increased incidence of RV dysfunction. A different structural response of microvessels to increased pressure load in the RV is a possible mechanism for this dysfunction. To examine the merit of this hypothesis, we explored the possibility that in the normal heart, the branching architecture of microvasculature in walls of the left ventricle (LV) and RV mature differently. The branching structure of intramyocardial arterioles and their downstream branches were investigated using three-dimensional (3D) micro-computed tomography (CT) images in different regions of the RV and LV walls of normal fetal, 1-mo, and 5-mo old pigs. The results point to a significant difference in the volume of myocardium perfused per vessel cross-sectional area (CSA) between the LV and RV walls at 5 mo. We speculate that this difference may be related to the reserve functional capacity of the LV, which requires a corresponding reserve in the expansion capacity of vasculature in the LV wall. PMID- 17426650 TI - Effect of television viewing at mealtime on food intake after a glucose preload in boys. AB - Television viewing (TVV) is considered a contributing factor to the development of childhood obesity yet it is unclear whether obesity results, in part, from increased energy intake during TVV. The objective of this study was to determine the effect of TVV on food intake (FI) of boys at a meal and its effect on caloric compensation at the test meal after a premeal glucose drink. On four separate mornings and in random order, boys received equally sweetened preloads containing Splenda sucralose or glucose [1.0 g/kg body weight (BW)] in 250 mL of water 2 h after a standard breakfast. Food intake from a pizza meal was measured 30 min later with or without TVV. Both preload treatment (p < 0.01) and TVV (p < 0.001) affected FI (kcal). TVV increased lunchtime FI by an average of 228 kcal. Glucose suppressed FI in the no TVV condition compared with control, but the effect was not statistically significant during TVV. Body composition and subjective appetite scores were positively associated with FI at the test lunch. In conclusion, TVV while eating a meal contributes to increased energy intake by delaying normal mealtime satiation and reducing satiety signals from previously consumed foods. PMID- 17426651 TI - The renin-angiotensin system in conscious newborn sheep: metabolic clearance rate and activity. AB - The role of the renin-angiotensin system (RAS) in regulating newborn mean arterial blood pressure (MAP) and tissue blood flow remains unclear. Although postnatal MAP increases, vascular responsiveness to infused angiotensin II (ANG II) is unchanged, possibly reflecting increased metabolic clearance rate of ANG II (MCR(ANG II)). To address this, we examined MAP, heart rate, plasma ANG II and renin activity (PRA), and MCR(ANG II) in conscious postnatal sheep (n = 9, 5-35 d old) before and during continuous systemic ANG II infusions to measure MCR (ANG II). Postnatal MAP increased (p < 0.02), whereas plasma ANG II decreased from 942 +/- 230 (SEM) to 471 +/- 152 and 240 +/- 70 pg/mL at <10 d, 10-20 d, and 21-35 d postnatally (p = 0.05), respectively. Despite high plasma ANG II, PRA remained elevated, averaging 6.70 +/- 1.1 ng/mL.h throughout the postnatal period, but decreased 35% (p = 0.01) during ANG II infusions. MCR(ANG II) decreased approximately sixfold after birth and averaged 115 mL/min.kg during the first month. Circulating ANG II is markedly increased after birth, reflecting placental removal, high fetal MCR(ANG II), and enhanced RAS activity. Although circulating ANG II decreases as MAP increases, MCR(ANG II) is unchanged, suggesting decreased ANG II production. Persistent vascular smooth muscle (VSM) AT2 receptor subtype (AT2R) expression after birth may modify the hypertensive effects of ANG II postnatally. PMID- 17426652 TI - Identification of a prosencephalic-specific enhancer of SALL1: comparative genomic approach using the chick embryo. AB - Comparative genomics is a promising approach for identifying regulatory elements governing the unique spatio-temporal expression patterns of morphogenetic genes. Conserved noncoding genomic sequences are candidate regulatory elements. Here we performed a survey for conserved noncoding elements (CNE) nested within the SALL1 gene; mutations in this gene result in the Townes-Brocks syndrome. A comparison of the genomic sequence between humans and chicken revealed five CNE. Genomic fragments corresponding to each CNE were inserted into reporter cassettes consisting of eGFP cDNA and a minimal promoter. These constructs were electroporated into chick embryos during gastrula, neurula, and pharyngula stages. Among the five CNE that were examined, one 443 bp CNE exhibited tissue specific enhancer activity. At the neurula stage, the eGFP signal was visualized in the prosencephalon. At the pharyngula stage, the eGFP signal was confined within the anterior neural ridge, which represents one of the morphogenetic centers regulating the patterning of the anterior neural plate. This report identifies, for the first time, an enhancer element of SALL1. PMID- 17426653 TI - Inhibition of nuclear factor-kappaB ameliorates bowel injury and prolongs survival in a neonatal rat model of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is a major cause of morbidity and death in premature infants. NEC is associated with increased levels of pro-inflammatory cytokines in plasma and tissues that are regulated by the transcription factor nuclear factor-kappaB (NF-kappaB). It remains unknown, however, whether NF-kappaB mediates injury in neonatal NEC. We therefore examined the activation status of NF-kappaB perinatally in the small intestine and in a neonatal rat model of NEC. We found that intestinal NF-kappaB is strongly activated at birth and, in dam-fed newborn rats, is down-regulated within a day. In contrast, NF-kappaB remains strongly activated at both d 1 and d 2 in stressed animals, and this is accompanied by a significant decrease in the levels of the endogenous NF-kappaB inhibitor protein IkappaBalpha and IkappaBbeta at d 2. To determine the importance of elevated NF-kappaB activity in intestinal injury in NEC, we administered the NEMO-binding domain (NBD) peptide that selectively inhibits the critical upstream IkappaB kinase (IKK). NBD but not a control peptide decreased mortality and bowel injury in this model, supporting the hypothesis that bowel injury in NEC results from elevated NF-kappaB activity. Our findings therefore lead us to conclude that selective NF-kappaB inhibition represents a promising therapeutic strategy for NEC. PMID- 17426654 TI - Interleukin-1beta, interleukin-18, and interferon-gamma expression in the cerebrospinal fluid of premature infants with posthemorrhagic hydrocephalus- markers of white matter damage? AB - Posthemorrhagic hydrocephalus (PHHC) represents a major complication of preterm birth. The aim of this study was to determine whether cerebrospinal fluid (CSF) levels of the pro-inflammatory cytokines IL-1beta, IL-18, and interferon (IFN) gamma are altered in the CSF of preterm infants with PHHC and may serve as a marker of white matter damage (WMD). Twenty-seven preterm infants with PHHC were included in the study; 13 of them had signs of cystic WMD (cWMD) on ultrasound examinations. CSF sample 1 was obtained at first ventriculostomy, sample 2 at shunt implantation. Results were compared with a control group of 20 age-matched patients without neurologic diseases. IL-1beta concentrations were elevated in CSF sample 1 of PHHC patients without WMD and in sample 1 of patients with cWMD. Concentrations of IL-18 were increased in both samples of patients without WMD and in sample 2 of patients with cWMD. CSF levels of IFN-gamma were elevated in sample 1 of PHHC patients with cWMD. The pro-inflammatory cytokine IL-1beta and IL-18 levels in the CSF are elevated in patients with PHHC. Higher IFN-gamma levels are detected in a subgroup of patients developing cWMD, indicating its involvement in the pathogenesis of cWMD in the context of PHHC. PMID- 17426655 TI - Pharmacokinetics of high-dose recombinant erythropoietin in plasma and brain of neonatal rats. AB - Recombinant human erythropoietin (rEpo) is neuroprotective in neonatal models of brain injury. Pharmacokinetic data regarding the penetration of circulating rEpo into brain tissue is needed to optimize neuroprotective strategies. We sought to determine the pharmacokinetics of rEpo given intraperitoneally or subcutaneously in plasma and brain. We hypothesized that 1) exogenous rEpo would penetrate the blood-brain barrier (BBB), 2) brain and plasma Epo would correlate, and 3) brain injury would enhance rEpo penetration. Two hundred and eighty-four 7-d-old control, sham, or brain-injured rats were treated with i.p. or s.c. rEpo (0, 250, 2500, or 5000 U/kg) and killed at scheduled intervals. Plasma and brain tissue were collected. Epo concentrations were measured by ELISA. Intraperitoneal injection yielded a faster and greater peak concentration of plasma rEpo (Tmax 3 h, Cmax 10,016 +/- 685 mU/mL) than s.c. injection (Tmax 9 h, Cmax 6224 +/- 753 mU/mL). Endogenous brain Epo was below detection even after hypoxia exposure. Systemic rEpo crossed the BBB in a dose-dependent manner, peaked in brain at 10 h, and was increased after brain injury. We conclude that high-dose rEpo is detectable in brain for >20 h after a single systemic injection. These pharmacokinetic data are valuable for planning of rEpo neuroprotection experiments. PMID- 17426656 TI - Variation in the normal hearing threshold predicts childhood IQ, linguistic, and behavioral outcomes. AB - Childhood hearing level varies considerably within the range considered normal. Four classes of outcome were investigated for associations with hearing thresholds in this range: ability to identify speech in noise, neurocognitive ability, linguistic ability, and behavior. The research was conducted in a general population cohort of 711 children with mean hearing threshold of 15 dB HL or better. Some outcomes: speech in noise, intelligence, and certain linguistic abilities, were predicted in both boys and girls; effects were stronger in girls. In girls only, poorer hearing predicted worse behavior. These effects remained after statistical control for childhood socioeconomic status and otitis media. Variability in normal hearing, due to causes other than otitis media, is associated with the listening, language, and neurocognitive abilities of children, and the behavior of girls. We suggest that these effects may be present for three reasons, cochlear insults, neurodevelopmental factors, and psychological factors. We discuss how these may interact to produce the effects observed. PMID- 17426657 TI - New predictors of the metabolic syndrome in children--role of adipocytokines. AB - There is ample discussion of the relevance of the metabolic syndrome, the definition criteria, and predictive power. Nevertheless, along with the increasing prevalence of childhood obesity, the prevalence of the metabolic syndrome in obese children is reported at 30%, irrespective of the definition applied. Because children are otherwise relatively free of co-morbidities, they constitute an interesting population in which to study the sequence of events of obesity-related pathology. The adipocytokines appear to be important in this respect. Leptin was initially suggested as a promising "antiobesity" hormone. New concepts indicate that, in humans, leptin and its soluble receptor may be more important in states of energy deficiency rather than a predictor of the metabolic syndrome. Adiponectin, on the other hand, is not only related to obesity and insulin resistance, but appears to be the strongest predictor for metabolic syndrome, even in children. In newborns and infants, both adipocytokines occur in high concentrations, even though this cannot completely explain the increased risk for ensuing metabolic disease later in life. Finally, low-grade systemic inflammation may underlie the clustering of metabolic risk factors, but their role in children remains to be specified. Overall factors from the adipose tissue may constitute not only markers but also mediators of metabolic sequelae of obesity. PMID- 17426658 TI - Elastolytic matrix metalloproteinases and coronary outcome in children with Kawasaki disease. AB - Kawasaki disease (KD) is a multisystem vasculitis that leads to coronary artery damage and aneurysm formation. Elastolytic matrix metalloproteinases (MMPs) have been implicated in the pathogenesis of arterial aneurysms. To determine the relationship between circulating levels of elastolytic MMPs and development of coronary artery aneurysms in children with KD, we partnered studies done in affected children with an animal model of disease. In affected children, circulating protein levels and enzymatic activity of MMP-2 and MMP-9 did not have a statistically significant relationship with coronary artery outcome after adjusting for demographic characteristics, and clinical and laboratory features. Although matrix-degrading proteolytic activity was specific for affected mice and localized to inflamed coronary artery segments, the enzymatic activity in the systemic circulation of affected and control mice were not different. Similar to affected children, peripheral blood levels of MMP-9 enzymatic activity did not correlate with coronary artery disease in the animal model. Therefore, circulating levels of MMPs known to act locally may not be useful biomarkers of disease. This is especially relevant to enzymatic activity that is tightly regulated at multiple levels including the local tissue environment. PMID- 17426659 TI - Effect of supplemental oxygen on reinitiation of breathing after neonatal resuscitation in rat pups. AB - To test our hypothesis that resuscitation with 21% and 40% oxygen (O2) would shorten time to onset of respiratory activity when compared with resuscitation with 100% O2, diaphragmatic electromyogram (EMG) electrodes were inserted in Sprague-Dawley rat pups, age 8-10 d before intubation and mechanical ventilation with 5% O2 to induce cessation of respiratory activity. Each animal was then resuscitated with 100% and 21% O2 (n = 10) or 100% and 40% O2 (n = 11) for 30 s before the ventilator was disconnected. Recovery of diaphragm activity was compared between resuscitation groups. Blood gas status and heart rate data were characterized in additional rat pups. Time to first respiratory effort was 36 +/- 21 s (mean +/- SD) for room air resuscitation and 72 +/- 22 s for 100% O2, (p = 0.002). In contrast, there was no difference in time to onset of diaphragm activity when resuscitation with 40% O2 was compared with 100% O2: 84 +/- 27 s versus 76 +/- 23 s, respectively (p > 0.05). Resuscitation with 100% and 40% O2 both resulted in hyperoxia and hypocapnia when compared with room air, without effect on heart rate. Our findings indicate that even modest hyperoxic resuscitation will result in a delayed onset of respiration compared with normoxic gas, via a mechanism that may involve both hyperoxemic and hypocapnic inhibition of chemoreceptors. PMID- 17426660 TI - Obesity, metabolic syndrome, and cardiovascular disease. AB - Early childhood overweight and obesity have alarmingly increased over the years. Adulthood obesity is a well demonstrated significant independent predictor of cardiovascular risk (CVR) and/or mortality, which predisposes to the major components of metabolic syndrome (MS). Evidence of MS in obese children has been also reported associated with biochemical and inflammatory factors that affect vascular physiologic function. Assessment of vascular function can be measured noninvasively in children allowing early detection of endothelial dysfunction and severe increase of arterial stiffness before clinical manifestations of atherosclerosis. Impairment of endothelial function related to the severity of obesity and to the degree of insulin resistance is considered as a condition that confers a premature atherogenicity status and is linked to adult conventional cardiovascular risk factors. Adipose tissue factors that interfere with insulin action and endothelial cell function have also been identified as major precursors of CVR factors. The metabolic and cardiovascular consequences of childhood obesity are well demonstrated and have a major impact on the development of atherosclerosis and lifetime CVR. The development of programs involving both diet and exercise for children with overt overweight/obesity appears to be essential to improve vascular function and metabolic disorders. Such interventions should be complemented by a primary prevention against childhood obesity. PMID- 17426661 TI - Obesity: do grapes hold the answer? PMID- 17426662 TI - Is your metabolism determined by (cell) fate? AB - The rise in the incidence of metabolic disease to become a major public health problem has been met with a substantial increase in research into both the clinical and basic science of metabolism. This work has revealed that the origins of metabolic diseases of adults can begin early in life. Furthermore, the age of onset of symptoms has been rapidly decreasing. Therefore, pediatricians should be critically involved in both the generation of new therapies as well as the institution of measures of disease prevention. This perspective examines how recent advances have improved our understanding of the development of metabolic diseases. A connection between glucocorticoids and the origins of metabolic disease is one enticing clue because of the clinical similarity between patients with glucocorticoid excess and those with metabolic disease. A unifying link was found by investigating the role of glucocorticoids on cell fate and differentiation of mesenchymal stem cells. We conclude that understanding the mechanisms by which glucocorticoids can modify how cell fate decisions are made holds promise for developing new therapies and preventative measures. PMID- 17426663 TI - Beyond the dopamine hypothesis to the NMDA glutamate receptor hypofunction hypothesis of schizophrenia. PMID- 17426664 TI - Serotonin syndrome associated with the use of escitalopram. AB - Escitalopram is the newest selective serotonin reuptake inhibitor (SSRI) available for use in the United States. It has been approved for the treatment of major depression and generalized anxiety disorder. It is the S-enantiomer of the SSRI citalopram and is highly serotonin specific as it has minimal effect on the reuptake of dopamine or norepinephrine. It is also a well-tolerated medication, with a side-effect profile comparable to the other SSRIs. While a number of side effects have been seen during escitalopram therapy, such as insomnia, nausea, and increased sweating, there are no reported cases of serotonin syndrome associated with escitalopram therapy to date. We present the case of a 24-year-old woman who developed serotonin syndrome after an increase in her escitalopram to 30 mg/day. We will review the diagnostic criteria of serotonin syndrome and the clinical scenarios in which serotonin syndrome can develop. We will also discuss the proposed treatments and role that polypharmacology may play in the development of this clinical entity. PMID- 17426665 TI - Balancing short-term symptom control and long-term functional outcomes in patients with Parkinson's disease. AB - Levodopa has played a central role in the treatment of Parkinson's disease for nearly 40 years and remains the single most effective symptomatic treatment for the disease. However, the response to levodopa therapy changes over time, and its long-term use is commonly associated with disabling motor complications. For this reason, the appropriate role of levodopa in the treatment of Parkinson's disease in particular, the question of when to initiate therapy with the drug-has been a matter of controversy. Because levodopa is the most effective treatment for Parkinson's disease, the management of this disease becomes a matter of balancing short-term symptom control with long-term functional outcomes. This article provides an overview of the basis for levodopa-associated motor complications and their impact on patients' clinical function and quality of life, followed by a discussion of strategies for managing these complications to achieve optimum symptom control while minimizing the adverse effects of long-term therapy. PMID- 17426666 TI - Mitochondrial myopathy complicated by eating disorder: a case report highlighting the potential interaction of genetic, metabolic, and psychodynamic factors. AB - A case is presented of a 56-year-old woman with a history of an eating disorder that preceded recognition of a mitochondrial myopathy. The possibility exists that her eating disorder was causally related to a more fundamental defect in mitochondrial oxidative metabolism. This case report highlights the phenotypic variability of mitochondrial myopathies. An increased risk of eating disorder may be associated with drugs that interfere with mitochondrial oxidative respiration. PMID- 17426667 TI - Level of insight and clinical features of obsessive-compulsive disorder with and without body dysmorphic disorder. AB - INTRODUCTION: Body dysmorphic disorder (BDD) and obsessive-compulsive disorder (OCD) have several similarities and are included among the obsessive-compulsive spectrum of disorders. However, the content of preoccupations and level of insight of BDD patients differ from OCD patients. OBJECTIVE: To compare the level of insight regarding obsessive-compulsive symptoms (OCS) and other clinical features in OCD patients with and without comorbid BDD. METHODS: We evaluated 103 OCD patients (n=25, comorbid BDD), according to Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria using the Structured Clinical Interview for DSM-IV, the Yale-Brown Obsessive-Compulsive Scale, the University of Sao Paulo Sensory Phenomena Scale, the Beck Depression and Anxiety Inventories, and the Brown Assessment of Beliefs Scale. RESULTS: The study groups differed significantly on several clinical features, including level of insight. A worse level of insight regarding OCS was independently associated with the presence of comorbid BDD. Lower educational level, more psychiatric comorbidities, presence of somatic and hoarding obsessions, and presence of intrusive images were associated with BDD comorbidity, even after adjusting for possible confounders. CONCLUSION: The presence of BDD in OCD patients is associated with poorer insight into obsessional beliefs and higher morbidity, reflected by lower educational levels and higher number of psychiatric comorbid disorders in general. PMID- 17426669 TI - History and prevalence of involuntary emotional expression disorder. AB - The syndrome now known as involuntary emotional expression disorder (IEED) is a condition characterized by uncontrollable episodes of laughing and/or crying. It has been known for more than a century, but confusing and conflicting terminology may have hampered the progress of physicians in recognizing this condition. IEED is associated with various neurological disorders and neurodegenerative diseases, including amyotrophic lateral sclerosis, multiple sclerosis, Parkinson's disease, Alzheimer's disease and other dementias, and neurological injuries such as stroke and traumatic brain injury. It is hoped that better defined terminology for IEED may help in the future diagnosis of this debilitating condition, the establishment of accurate prevalence rates for IEED in the varying underlying conditions, and also in removing blame and stigma from sufferers by providing reassurance about the nature of their condition. PMID- 17426670 TI - Involuntary emotional expression disorder: definition, diagnosis, and measurement scales. AB - Involuntary emotional expression disorder (IEED) is a syndrome in which emotional or affective motor control becomes dysregulated as a result of brain damage from neurological disease or as a result of brain injury. A debilitating disorder with persistent symptoms, IEED has a significant impact upon the lives of patients and caregivers, but is often overlooked or misdiagnosed. IEED is a clinically well defined disorder, and specific criteria may be used when making a diagnosis. It must be distinguished from depression or other psychological and neurological conditions. Several assessment scales have been validated for use in IEED, in order to evaluate the baseline severity and to track the course of the disorder and response to treatment. PMID- 17426671 TI - Pathophysiology of involuntary emotional expression disorder. AB - Extensive clinical experience and research suggest that a cortico-limbic subcortico-thalamic-ponto-cerebellar network plays a significant role in the expression of human emotions. This network includes specific cerebral, cerebellar, and brainstem areas and their multiple projections/pathways, with activity modulated through serotonergic, dopaminergic, glutamatergic, and possibly sigma receptor neurotransmitter systems. Disruptions of regulatory and inhibitory mechanisms in the structure and function of this network likely constitute a pathophysiological basis for the crying and laughing episodes characteristic of involuntary emotional expression disorder. Pharmacologic interventions targeting the neurochemical modulators of the emotional expression systems may afford opportunities for symptom control among persons affected by this disorder. PMID- 17426672 TI - Involuntary emotional expression disorder: treating the untreated. AB - Patients with involuntary emotional expression disorder (IEED) have impaired social and occupational functioning and there is currently no Food an Drug Administration-approved treatment. Treatment options include tricyclic antidepressants (TCAs), selective serotonin reuptake inhibitors (SSRIs), dopaminergic agents, and a combination of dextromethorphan and quinidine. Studies of monaminergic agents have typically been small and executed in single-center settings. Assessment measures generally show significant symptomatic improvements, including a reduction in the number of laughing or crying episodes and improvements in patients' clinical condition. The tolerability profiles of these agents are well defined, and include dizziness, tachycardia and QTc prolongation (TCAs), and sleep and sexual disturbances (SSRIs). The combination of dextromethorphan and quinidine has also been assessed in two large multicenter studies in patients with amyotrophic lateral sclerosis and multiple sclerosis. Compared with placebo and either agent alone, there were significant improvements in symptoms, quality of life, and relationships. The most common side effects were dizziness and nausea, and potential drug interactions with quinidine should also be considered. Choice of treatment should be evidence-based, taking into account both efficacy and tolerability. PMID- 17426673 TI - ADHD: prevalence, diagnosis, and issues of comorbidity. PMID- 17426674 TI - [Echocardiographic assessment of results of single stage mitral and aortic valvuloplasty]. AB - Echocardiographic assessment of results of single stage mitral and aortic valvuloplasty was carried out in 29 patients (mean age 44.1+/-1.2 years) with combined rheumatic mitral and aortic valve disease. Dynamics of most of ultrasound parameters of the heart after double intervention on mitral and aortic valves was positive. Aortic regurgitation was either absent or minimal. Mean pressure gradient across the aortic valve was reduced 50%. Average reduction of transmitral valve pressure gradient was > or = 70%. Average increase of mitral orifice area index was 146%; average decrease of left atrial diameter index was 17%. PMID- 17426675 TI - [Visceral obesity as a pathogenetic factor of ischemic heart disease]. AB - Relationship between visceral obesity and development of ischemic heart disease (IHD) has been widely studied during last years. Fat tissue is considered to be endocrine and secretory organ liberating wide spectrum of protein and signal factors-adipokines. We studied 27 healthy subjects and 55 patients (including 39 patients with IHD) with visceral and gynoid obesity. Abdominal fat content was assessed quantitatively by computed tomography. It was proven that total volume of fat tissue in visceral obesity is substantially elevated especially in mesogastrium, omentum, and around portal vein, where increased amounts of very low density lipoproteins, free fatty acids, and low density lipoproteins are generated. Enhanced inflow of free fatty acids and glucose to liver constitutes pathophysiologic basis of visceral obesity. This very process creates elevation of optical density of tissue in visceral obesity. Elevation of low density lipoproteins, very low density lipoproteins, and cholesterol are important factors in pathogenesis of IHD. PMID- 17426676 TI - [Peculiarities of regulatory mechanisms of autonomic nervous system in patients with arterial hypertension and disturbed 24-hour blood pressure profile]. AB - BACKGROUND: Insufficient nocturnal blood pressure (BP) decline is associated with elevated risk of complications of hypertensive disease. Heart rate variability (HRV) reflects activity of sympathetic and parasympathetic parts of autonomic nervous system. AIM: To elucidate special characteristics of HRV in patients with various types of 24-hour BP rhythm. Material and methods. Bifunctional 24-hour monitoring and echocardiography were carried out in 42 men with stage I-II hypertensive disease and I-II degree of arterial hypertension (mean age 21.7+/ 4.5 years) and 16 practically healthy young people (mean age 24.6+/-5.2 years). RESULTS: Subjects with insufficient (<10%) and adequate nocturnal BP decline (non dippers and dippers) were distinguished (groups ND and D, respectively). Patients with hypertension in group ND had elevation of systolic BP variability during night and day time, augmentation of nocturnal and diurnal HRV low frequency power, lowering of nocturnal high frequency power, lowering of pNN50 and rMSSD values at night. HRV parameters of control subjects in this group did not differ from those of healthy people. All HRV parameters in group D were characterized by significant 24-hour rhythmicity. This rhythmicity was substantially disturbed in patients of group ND. Parameters of central hemodynamics were similar in groups D and ND. CONCLUSION: These results evidence for the presence of enhanced activity of sympathetic part of autonomic nervous system in non-dipper patients with hypertensive disease throughout 24 hours and during night time and for disturbed circadian rhythm of autonomic nervous system activity. This can serve as a basis for increased rate of cardiovascular complications in this category of patients. PMID- 17426677 TI - [Prevention of ischemic stroke in middle aged patients with atrial fibrillation. Effect of sinus rhythm maintenance, aspirin, warfarin, and simvastatin]. AB - Patients (n=263, age 49-64 years) with persistent nonvalvular atrial fibrillation were subjected to controlled treatment aimed at restoration and maintenance of sinus rhythm (n=134) or slowing of rate of ventricular contractions. Efficacy of complementary use of combination of simvastatin with aspirin, warfarin and aspirin for prevention of ischemic stroke was assessed in both of these strategies. First strategy, supplemented with combination of simvastatin and aspirin prevented ischemic stroke and lethal outcome most successfully, while the use of warfarin was unexpectedly associated with comparative elevation of risk of ischemic stroke. It is impossible to exclude that long term administration of warfarin in strategy of sinus rhythm maintenance attenuated advantages of normal rhythm in relation of lowering of stroke risk in AFFIRM study and other similar works. Contrary to known cardio-embolic mechanism of cerebral embolism in permanent atrial fibrillation main aim of the treatment in successful sinus rhythm maintenance may become prevention of arterio-arterial emboli. PMID- 17426678 TI - [Effect of corticosterone and plasma lipoproteins on working capacity and ultrastructure of isolated rat heart]. AB - Blood lipoproteins (very low density--VLDL, low density--LDL and high density- HDL) penetrate into the myocardium through capillary endothelial layer via receptor mediated endocytosis (all lipoproteins), nonreceptor uptake, or along interendothelial gaps (LDL). In the myocardium lipoproteins are captured by interstitial macrophages and are subjected to degradation in secondary liposomes. Their action on myocardium results in development of perivascular swelling and constriction of substantial portion of capillaries. However part of capillaries (about 20%) stay in a condition of vasodilation. Destructive changes revealed (myelin figures, mild lysis of myofibrils) are presumably caused by activation of lysosomal proteinases. Corticosterone lowered coronary flow velocity, while parameters of working capacity of the heart remained at control level. Combined use of corticosterone and VLDL suppressed myocardial functional activity, to a great extent because of diminishment of coronary flow velocity. Corticosterone and LDL exerted less pronounced negative effect. Corticosterone and HDL caused improvement of parameters of cardiac working capacity. PMID- 17426679 TI - [Bone marrow stem cells in the treatment of ischemic heart disease]. AB - Cardiac cells possess limited ability to regeneration. Therefore acute myocardial infarction results in replacement of "perished" cardiomyocytes by scar tissue. The consequence of this is lowering of myocardial contractile function and development of heart failure. Novel approach to improvement of blood supply, contractility and possibly heart failure prevention--transplantation of stem cells--has attracted increasing attention during recent years. The use of autologous bone marrow stem cells is one of most perspective directions of cellular therapy. Until present time mechanism of action of stem cells, necessary amount of cellular material, most adequate method of stem cells introduction, methods of diagnosis of clinical efficacy of cellular therapy have not been fully elucidated. PMID- 17426680 TI - [Inflammation and chronic heart failure. The role of statins]. AB - Immune inflammation is an important link in pathogenesis of chronic heart failure (CHF). It has been proven that hyperproduction of proinflammatory cytokines (TNFa, IL-6, IL-1, etc) with associated endothelial dysfunction and oxidative stress affect unfavorably clinico-hemodynamic parameters and prognosis of life of patients. However attempt to augment efficacy of treatment of CHF by means of inclusion of TNFa activity inhibitors in the complex of main remedies turned out unsuccessful. At present under discussion are perspectives of application in this category of patients of statins--lipid lowering drugs with pleiotropic properties most important of which is antiinflammatory action. Theoretical obstacles for the use of statins constitute available epidemiological data on reverse relationship between cholesterol level and mortality of patients with CHF in a framework of endotoxin-lipoprotein theory. At the same time preclinical and clinical experience has been accumulated evidencing for perspectiveness of such approach. Final solution of the problem of safety and feasibility of application of statins in CHF is expected after completion of prospective randomized trials GISSI-HF and CORONA. PMID- 17426681 TI - [Aneurysm of internal carotid artery. Clinical symptoms, surgical treatment, results]. AB - Aneurysms of the extracranial portion of the internal carotid artery is rare pathology - only 2.2% of all surgical procedures on this portion of the internal carotid artery are caused by this disease. Ten-year experience in surgical treatment of these aneurysms is presented in article. Symptoms and etiology of this disease are considered. It is demonstrated that fibromuscular dysplasia is one of the main causes of these aneurysms. Strategy and policy of surgical treatment, clinical cases are analyzed. Resection of the aneurysm with angioplasty or grafting is optimal. End-to-end anastomosis is possible in patients with kinking. Results of surgical treatment are positive, there is no hospital lethality. PMID- 17426682 TI - [Preoperative preparation of patients with stage IV of atherosclerotic chronic lower limb ischemia]. AB - Results of surgical treatment of 322 patients with stage IV atherosclerotic chronic lower limb ischemia are discussed. Among the patients not treated with specific therapy for prevention of postoperative complications the lethality was 17.3%. Careful examination before surgery and preventive measures permitted to reduce lethality to 11%. PMID- 17426683 TI - [Morphological characteristics and intracellular electrolytic composition of blood cells at surgeries on lower extremities arteries]. AB - Morphological characteristics and trace and macroelement composition of blood cells flowing out of ischemic lower limb before, during and after reconstructive surgeries under different type of anesthesia. A total of 102 male patients aged 45 to 60 years with atherosclerotic occlusions of the arteries of the femoral popliteal zone were included into the study. According to anesthesia type all the patients were divided into 3 groups: group 1 consisted of 34 patients operated under spinal anesthesia, 37 patients of group 2 underwent surgery under combined anesthesia (spinal anesthesia with intravenous sedation), 31 patients of group 3 under total intravenous anesthesia with myoplegia and artificial pulmonary ventilation. All the blood examinations were carried out with scanning electron microscope XL-30 ("Philips") and X-ray spectrum microanalyzer Edax ( "Edax International", USA). It is demonstrated that in spinal and combined anesthesia morphological characteristics of blood cells normalized due to optimization of intraerythrocytic and intrathrombocytic electrolytic homeostasis unlike total intravenous anesthesia, when intracellular imbalance of trace and macroelements progresses and ultrastructural cellular alterations persist. PMID- 17426684 TI - [Is antisecretory therapy of patients with chemical burn of the esophagus mandatory?]. AB - Intragastric pH-metry and esophagogastroduodenoscopy has been performed in 64 patients with chemical burn of the esophagus. It has been revealed that 51.6% patients had hyperacidity, 25% - normacidity, and 23.4% - hypoacidity. Gastroesophageal reflux has been diagnosed in 50% patients according to pH-metry examination and in 80-100% patients according to esophagogastroduodenoscopy, duodenogastric reflux - in 25 and 34.4% cases, respectively. A direct correlation exists between gastroesophageal reflux and basal gastric acidity, severity of chemical burn of the esophagus. It is concluded that antisecretory therapy is mandatory in complex treatment of patients with chemical burn of the esophagus. PMID- 17426685 TI - [Surgical treatment of cicatricial stricture of the esophagus and stomach after chemical burn]. AB - The experience with treatment of 104 patients with burn cicatricial strictures of the esophagus and stomach is analyzed. Postoperative complications were seen in 28 (26.9%) patients. Hospital lethality was 12.5%. It is concluded that method of esophagoplasty in these patients should be selected individually. PMID- 17426686 TI - [Syndrome of intraabdominal hypertension in patients with destructive forms of pancreatitis]. AB - Results of intraabdominal pressure examination in 129 patients with acute destructive pancreatitis are presented. It is demonstrated that pancreonecrosis was associated with intraabdominal pressure rise in 78% cases, and intraabdominal hypertension syndrome developed in 9.3% patients. Prolonged intraabdominal hypertension in patients with advanced pancreonecrosis requires laparotomy. Correlation between intraabdominal hypertension and diffusion of pancreatogenic intra- and retroabdominal inflammation, and also between intraabdominal hypertension and condition severity according to APACHE II scale is demonstrated. PMID- 17426687 TI - [Assessment of severity assessment and prognosis of pyodestructive abdominal diseases (with editor's comment)]. AB - Leukocytic index of intoxication (LII), number of leukocytes and lymphocytes in blood were compared with clinical symptoms and histological examination of removed organs in patients with acute pyodestructive diseases of abdominal organs. Correlation between LII and inflammation severity was revealed. It is also demonstrated that high LII with low level of lymphocytes is a negative prognostic sign. PMID- 17426688 TI - [Polyposis of the stomach. Endoscopic treatment or follow-up?]. AB - Out of 3243 patients with gastric polips, polyposis was diagnosed in 298 ones including 135 cases of complicated and 163 cases of uncomplicated forms. Hyperplastic polips were diagnosed in 281 patients. Along with hyperplastic polyps, solitary adenomas were diagnosed in 9 cases, Peitz-Jegerc polyps - in 6, and juvenile polyps - in 2. Polypectomy was performed in 153 patients: partial - in 104, and total - in 49. Follow-up was carried out in 98 non-operated patients, indications for endoscopic surgery were seen in 18 cases. After partial endoscopic polypectomy, repolypectomy was performed in 24 cases: single - in 8, multiple - in 16. Repeated endoscopic treatment after total polypectomy was performed in 3 patients. There were no cases of polyp transformation into cancer. Open surgical treatment was not performed. There were no severe complications after endoscopic treatment. PMID- 17426689 TI - [Recurrences after surgical treatment of early (pT1) cancer of the stomach: laws of development, extended lymphadenectomy in prophylaxis of recurrences]. AB - Long-term results of treatment of 175 patients with early cancer of the stomach are analyzed. Recurrences of the disease (local, regional recurrences and distant metastases) were diagnosed in 14 (8.2% of all operated) patients. Duration of recurrence-free period ranged from 2-3 months to 9.5 years; median was 12 months. Tumor spread, regional nodes affection, tumor morphological structure, age of patients were the main prognostic factors for recurrences. Extent of surgery and lymphodissection did not correlate with recurrences rate. It is concluded that extended lymphodissection in patients with early cancer of the stomach should not be regarded as a real method for an increase of treatment efficacy. PMID- 17426690 TI - [Purulent and necrotic complications of diabetic foot syndrome and their reflection in ICD-10 and standards of insurance medicine]. AB - Treatment of 1046 patients with complicated forms of diabetic foot is analyzed. New classification of purulent and necrotic complication of diabetic foot is described. Current medical-economic standards for diabetic foot complicated forms are subjected to criticism. A new concept of the medical-economic standard is discussed. PMID- 17426691 TI - [The role of a surgical method in combined and complex treatment of patients with squamous cell oral and oropharyngeal cancer]. AB - Surgery was used in the treatment of 194 patients with tumors of the oral cavity and throat. In 110 of them the treatment was combined (radiotherapy and surgery) and in 84 - complex (polychemotherapy + surgery + postoperative radiotherapy). Characteristics of surgeries are compared. It is demonstrated that number of function-preserving operations in patients given complex treatment was 86.9%, combined treatment - 70.9%. PMID- 17426692 TI - [Prophylaxis and treatment of primary insufficiency of bronchial stump after pneumonectomy]. AB - Efficacy of gel DAM+ in bronchial surgery was evaluated in experimental and clinical studies. In clinical practice DAM+ was used for treatment of 22 patients with insufficiency of bronchial stump, and in 12 patients with high risk of stump insufficiency this gel was administered into submucosal layer before stump closure. It is demonstrated that administration of DAM+ gel into submucosal layer is simple and effective method of treatment and prophylaxis of bronchial stump insufficiency. PMID- 17426693 TI - In vitro derivation of functional insulin-producing cells from human embryonic stem cells. AB - The capacity for self-renewal and differentiation of human embryonic stem (ES) cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus. Here, we report a newly developed and effective method, carried out in a serum-free system, which induced human ES cells to differentiate into insulin-producing cells. Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells, as detected by the expression of the definitive endoderm markers Sox17 and Brachyury. Further, all-trans retinoic acid (RA) was used to promote pancreatic differentiation, as indicated by the expression of the early pancreatic transcription factors pdx1 and hlxb9. After maturation in DMEM/F12 serum-free medium with bFGF and nicotinamide, the differentiated cells expressed islet specific markers such as C-peptide, insulin, glucagon and glut2. The percentage of C-peptide-positive cells exceeded 15%. The secretion of insulin and C-peptide by these cells corresponded to the variations in glucose levels. When transplanted into renal capsules of Streptozotocin (STZ)-treated nude mice, these differentiated human ES cells survived and maintained the expression of beta cell marker genes, including C-peptide, pdx1, glucokinase, nkx6.1, IAPP, pax6 and Tcf1. Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia; and the corrected phenotype was sustained for more than six weeks. Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus. PMID- 17426694 TI - A nonsynonymous SNP in human cytosolic sialidase in a small Asian population results in reduced enzyme activity: potential link with severe adverse reactions to oseltamivir. AB - The use of oseltamivir, widely stockpiled as one of the drugs for use in a possible avian influenza pandemic, has been reported to be associated with neuropsychiatric disorders and severe skin reactions, primarily in Japan. Here we identified a nonsynonymous SNP (single nucleotide polymorphism) in dbSNP database, R41Q, near the enzymatic active site of human cytosolic sialidase, a homologue of virus neuraminidase that is the target of oseltamivir. This SNP occurred in 9.29% of Asian population and none of European and African American population. Our structural analyses and Ki measurements using in vitro sialidase assays indicated that this SNP could increase the unintended binding affinity of human sialidase to oseltamivir carboxylate, the active form of oseltamivir, thus reducing sialidase activity. In addition, this SNP itself results in an enzyme with an intrinsically lower sialidase activity, as shown by its increased Km and decreased Vmax values. Theoretically administration of oseltamivir to people with this SNP might further reduce their sialidase activity. We note the similarity between the reported neuropsychiatric side effects of oseltamivir and the known symptoms of human sialidase-related disorders. We propose that this Asian enriched sialidase variation caused by the SNP, likely in homozygous form, may be associated with certain severe adverse reactions to oseltamivir. PMID- 17426695 TI - Hormones and cancer. PMID- 17426696 TI - EMT or apoptosis: a decision for TGF-beta. PMID- 17426697 TI - PTEN, a general negative regulator of cyclin D expression. PMID- 17426698 TI - A novel function of p53 in TR3-MDM2 cross-talk. PMID- 17426699 TI - Ewings family oncoproteins: drunk, disorderly and in search of partners. PMID- 17426700 TI - Back to the future of oridonin: again, compound from medicinal herb shows potent antileukemia efficacies in vitro and in vivo. PMID- 17426701 TI - Offspring sex ratio and gonadal irradiation in the British Childhood Cancer Survivor Study. AB - We investigated offspring sex ratio among 6232 offspring born to 3218 survivors of childhood cancer in relation to therapeutic irradiation, and pooled our data with those from two other large-scale studies giving a total of 9685 offspring. Exposure to high-dose gonadal irradiation was not associated with a significant alteration in offspring sex ratio compared to low doses (men: P=0.58, women: P=0.66). There was also no evidence that the ratio varied with time since cancer diagnosis when comparing survivors treated with radiotherapy vs those without (men: P=0.51; women: P=0.46). This, the largest study to date, finds no evidence that exposure to radiation affects the offspring sex ratio among survivors of childhood cancer. PMID- 17426702 TI - Combination treatment with Grb7 peptide and Doxorubicin or Trastuzumab (Herceptin) results in cooperative cell growth inhibition in breast cancer cells. AB - Grb7 has potential importance in the progression of cancer. We have previously identified a novel peptide that binds to the SH2 domain of Grb7 and inhibits its association with several different receptor tyrosine kinases. We have synthesised the Grb7 peptide, G7-18NATE, with two different cell penetrating peptides, Penetratin and Tat. In this study, we have shown that both Penetratin- and Tat conjugated G7-18NATE peptides are able to inhibit the proliferation of SK-BR-3, ZR-75-30, MDA-MB-361 and MDA-MB-231 breast cancer cells. There was no significant effects on breast cancer MCF-7cells, non-malignant MCF 10A or 3T3 cells. In addition, there was no significant inhibition of proliferation by Penetratin or Tat alone or by their conjugates with arbitrary peptide sequence in any of the cell lines tested. We determined the EC50 of G7-18NATE-P peptide for SK-BR-3 cell proliferation to be 7.663 x 10(-6) M. Co-treatment of G7-18NATE-P peptide plus Doxorubicin in SK-BR-3 breast cancer cells resulted in an additional inhibition of proliferation, resulting in 56 and 84% decreases in the Doxorubicin EC50 value in the presence of 5 x 10(-6) and 1.0 x 10(-5) M G7-18NATE-P peptide, respectively. Importantly, the co-treatment with Doxorubicin and the delivery peptide did not change the Doxorubicin EC50. Since Grb7 associates with ErbB2, we assessed whether the peptide inhibitor would have a combined effect with a molecule that targets ErbB2, Herceptin. Co-treatment of Herceptin plus 1.0 x 10( 5) M G7-18NATE-P peptide in SK-BR-3 cells resulted in a 46% decrease in the Herceptin EC50 value and no decrease following the co-treatment with Herceptin and penetratin alone. This Grb7 peptide has potential to be developed as a therapeutic agent alone, in combination with traditional chemotherapy, or in combination with other targeting molecules. PMID- 17426703 TI - Reproductive risk factors for endometrial cancer among Polish women. AB - We conducted a population-based case-control study of reproductive factors in Warsaw and Lodz, Poland, in 551 incident endometrial cancer cases and 1925 controls. The reproductive variable most strongly related to risk was multiparity, with subjects with three or more births having a 70% lower risk than the nulliparous women. The reduced risk was particularly strong below 55 years of age. Subjects with older ages at a first birth were also at reduced risk even after adjustment for number of births. Ages at last birth or intervals since last birth were not strongly related to risk. Spontaneous abortions were unrelated to risk, but induced abortions were associated with slight risk increases (odds ratios=1.28, 95% confidence intervals 0.8-2.1 for 3+ vs no abortions). The absence of effects on risk of later ages at, or short intervals since, a last birth fails to support the view that endometrial cancer is influenced by mechanical clearance of initiated cells. Alternative explanations for reproductive effects should be sought, including alterations in endogenous hormones. PMID- 17426704 TI - Studies on p53, BAX and Bcl-2 protein expression and microsatellite instability in stage III (UICC) colon cancer treated by adjuvant chemotherapy: major prognostic impact of proapoptotic BAX. AB - We evaluated the expression patterns of proapoptotic BAX, antiapoptotic Bcl-2 and p53, the proposed upstream effector of these molecules, as potential prognostic markers in UICC stage III colon cancer by immunohistochemical staining. To identify high-frequency microsatellite instability (MSI+) individuals, we performed single-strand conformation polymorphism-based analysis for BAT26. A total of 188 patients who had received 5-fluorouracil (5-FU)-based adjuvant chemotherapy (5-FU/folinic acid or 5-FU/levamisole) were enrolled. Median follow up was 84.5 months. We found that BAX, Bcl-2 and p53 protein expressions were high or positive in 59, 70 and 50% of 188 cases, respectively. MSI+ tumours were detected in 9% of 174 evaluable patients. BAX or Bcl-2 was correlated with a higher degree of differentiation or left-sided tumours (P=0.01 or P=0.03, respectively); MSI was correlated with right-sided tumours (P<0.0001). In contrast to p53, Bcl-2, or MSI, low BAX, advanced pN category, low grade of differentiation and treatment with 5-FU/levamisole were univariately associated with poorer disease-free survival (DFS) (P=0.0005, P=0.001, P=0.005 and P=0.01, respectively) and poorer overall survival (OS) (P=0.002, P=0.0001, P=0.003 and P=0.02, respectively). Besides pN category and treatment arm, BAX was an independent variable related to both OS and DFS (P=0.003 and P=0.001, respectively). In both univariate and multivariate analysis, the p53-/BAX high in comparison with the p53+/BAX high subset conferred a significantly improved DFS (P=0.03 and P=0.03, respectively) as well as a marginally improved OS (P=0.07 and P=0.08, respectively). BAX protein expression may be of central significance for clinical outcome to 5-FU-based adjuvant chemotherapy in stage III colon cancer, and bivariate analysis of p53/BAX possibly may provide further prognostic evidence. PMID- 17426705 TI - In vivo inhibition of angiogenesis by sulphamoylated derivatives of 2 methoxyoestradiol. AB - Drugs that inhibit growth of tumours and their blood supply could have considerable therapeutic potential. 2-Methoxyoestradiol-3,17-O,O-bis-sulphamate (2-MeOE2bisMATE) has been shown to inhibit the proliferation of MCF-7 (ER+) breast cancer cells and angiogenesis in vitro. 2-MeOE2bisMATE and its analogue, 17-Cym-2-MeOE2MATE, were investigated for their ability to inhibit in vivo angiogenesis and tumour growth. The mouse Matrigel plug assay for angiogenesis was used to investigate the effect of compounds on neovascularisation and was quantified using a FITC-dextran injection technique. Nude mice bearing tumours derived from MCF-7 cells were used to assess efficacy on tumour growth. Tumour sections were stained for VEGFR-2 and Ki67 to assess tumour angiogenesis and cell proliferation respectively. Matrigel plugs supplemented with basic fibroblast growth factor resulted in increased neovascularisation over 7 days. Oral administration of 2-MeOE2bisMATE for 7 days at 10 or 50 mg kg(-1) significantly reduced neovascularisation to or below control levels respectively. 17-Cym-2 MeOE2MATE at 20 mg kg(-1) was equally effective. 2-MeOE2bisMATE, dosed daily for 21 days, caused a 52% reduction in tumour growth at 5 mg kg(-1) and 38% regression at 20 mg kg(-1). 17-Cym-2-MeOE2MATE (20 mg kg(-1)) reduced tumour growth by 92%. Immunohistochemistry revealed a reduction in angiogenesis and proliferation. Matrigel plug and tumour imaging after FITC-dextran injection indicated that 2-MeOE2bisMATE caused a marked disruption of vasculature. These sulphamoylated oestrogen derivatives have been shown to be potent inhibitors of angiogenesis in vivo. This, together with their ability to inhibit tumour growth, indicates the potential of this new class of drugs for further development for cancer therapy. PMID- 17426706 TI - A preclinical evaluation of pemetrexed and irinotecan combination as second-line chemotherapy in pancreatic cancer. AB - Gemcitabine (GEM)-based chemotherapy is regarded as the standard treatment of pancreatic adenocarcinoma, but yields a very limited disease control. Very few studies have investigated salvage chemotherapy after failure of GEM or GEM containing chemotherapy and preclinical studies attempting to widen the therapeutic armamentarium, not including GEM, are warranted. MIA PaCa2, CFPAC-1 and Capan-1 pancreatic cancer cell lines were treated with GEM, fluouracil (5 FU), docetaxel (DCT), oxaliplatin (OXP), irinotecan (CPT-11), pemetrexed (PMX) and raltitrexed (RTX) as single agent. Pemetrexed, inducing apoptosis with IC50s under the Cmax in the three lines tested, appeared the most effective drug as single agent. Based on these results, schedule- and concentration-dependent drug interactions (assessed using the combination index) of PMX/GEM, PMX/DCT and PMX CPT-11 were evaluated. The combinatory study clearly indicated the PMX and CPT-11 combination as the most active against pancreatic cancer. To confirm the efficacy of PMX-CPT-11 combination, we extended the study to a panel of 10 pancreatic cancer cell lines using clinically relevant concentrations (PMX 10 microM; CPT-11 1 microm). In eight of 10 lines, the PMX-CPT-11 treatment significantly reduced cell recovery and increased both the subG1 and caspase 3/7 fraction. After a 5 day wash out period, an increased fraction of subG1 and caspase3/7 persisted in PMX-CPT-11-pretreated cell lines and a significant reduction in the clonogenicity capacity was evident. Finally, in vivo, the PMX/CPT-11 combination showed the ability to inhibit xenograft tumours growth as second-line therapy after GEM treatment. The PMX and CPT-11 combination displays a strong schedule-independent synergistic cytotoxic activity against pancreatic cancer, providing experimental basis for its clinical testing as salvage chemotherapy in pancreatic cancer patients. PMID- 17426707 TI - No efficacy of annual gynaecological screening in BRCA1/2 mutation carriers; an observational follow-up study. AB - BRCA1/2 mutation carriers are offered gynaecological screening with the intention to reduce mortality by detecting ovarian cancer at an early stage. We examined compliance and efficacy of gynaecological screening in BRCA1/2 mutation carriers. In this multicentre, observational, follow-up study we examined medical record data of a consecutive series of 888 BRCA1/2 mutation carriers who started annual screening with transvaginal ultrasonography and serum CA125 between 1993 and 2005. The women were annually screened for 75% of their total period of follow up. Compliance decreased with longer follow-up. Five of the 10 incident cancers were interval tumours, diagnosed in women with a normal screening result within 3 10 months before diagnosis. No difference in stage distribution between incident screen-detected and interval tumours was found. Eight of the 10 incident cancers were stage III/IV (80%). Cancers diagnosed in unscreened family members had a similar stage distribution (77% in stage III/IV). The observed number of cases detected during screening was not significantly higher than expected (Standardized Incidence Ratio (SIR): 1.5, 95% confidence interval: 0.7-2.8). For the subgroup that was fully compliant to annual screening, a similar SIR was found (1.6, 95% confidence interval: 0.5-3.6). Despite annual gynaecological screening, a high proportion of ovarian cancers in BRCA1/2 carriers are interval cancers and the large majority of all cancers are diagnosed in advanced stages. Therefore, it is unlikely that annual screening will reduce mortality from ovarian cancer in BRCA1/2 mutation carriers. PMID- 17426708 TI - WTH3 is a direct target of the p53 protein. AB - Previous results showed that overexpression of the WTH3 gene in multidrug resistance (MDR) cells reduced MDR1 gene expression and converted their resistance to sensitivity to various anticancer drugs. The WTH3 gene promoter was found to be differentially regulated in paired MDR vs non-MDR MCF7 cells owing to epigenetic modifications and transcription factor modulations. To understand further the mechanisms that govern WTH3's differential expression, we uncovered a p53-binding site in its promoter, which indicated that WTH3 could be regulated by the p53 gene. This hypothesis was then tested by different strategies. The resulting data revealed that (1) the WTH3 promoter was upregulated by the p53 transgene in diverse host cells; (2) there was a correlation between WTH3 expression levels and p53 gene status in a cell line panel; (3) a WTH3 promoter region was directly targeted by the p53 protein in vitro and in vivo. In addition, overexpression of the WTH3 gene promoted the apoptotic phenotype in host cells. On the basis of these findings, we believe that the negative role played by the WTH3 gene in MDR development is through its proapoptotic potential that is regulated by multiple mechanisms at the transcription level, and one of these mechanisms is linked to the p53 gene. PMID- 17426709 TI - Targeted Sleeping Beauty transposition in human cells. AB - Transposons are natural gene delivery vehicles. The Sleeping Beauty (SB) transposon shows efficient transposition and long-term transgene expression in the cells of vertebrates including humans. SB transposition into chromosomal DNA occurs in a fairly random manner. This is clearly not desirable in human gene therapeutic applications because there are potential genotoxic effects associated with transposon integration. In this study we set out to manipulate the selection of SB's target sites for targeted transposition into predetermined chromosomal regions. We evaluated experimental strategies based on engineered proteins composed of DNA-binding domains fused to (i) the transposase; (ii) another protein that binds to a specific DNA sequence within the transposable element; and (iii) another protein that interacts with the transposase. We demonstrated targeted transposition into endogenous matrix attachment regions (MARs) and a chromosomally integrated tetracycline response element (TRE) in cultured human cells, using targeting proteins that bind to the transposon DNA. An approach based on interactions between the transposase and a targeting protein containing the N-terminal protein interaction domain of SB was found to enable an approximately 10(7)-fold enrichment of transgene insertion at a desired locus. Our experiments provide proof-of-principle for targeted chromosomal transposition of an otherwise randomly integrating transposon. Targeted transposition can be a powerful technology for safe transgene integration in human therapeutic applications. PMID- 17426710 TI - Role of acid ceramidase in resistance to FasL: therapeutic approaches based on acid ceramidase inhibitors and FasL gene therapy. AB - Head and neck squamous cell cancers (HNSCC) are particularly aggressive and are resistant to many forms of treatment. Ceramide metabolism has been shown to play an important role in cancer progression and cancer resistance to therapy in many tumor models, including HNSCC. Here, we study the role of the ceramide metabolizing enzyme acid ceramidase (AC) in therapeutic responses in HNSCC. First, we show that AC is over-expressed in 70% of head and neck squamous cell tumors compared with normal tissues, suggesting that this enzyme may play an important role in facilitating HNSCC growth. Next, comparison of three HNSCC cell lines with low, medium, and high levels of AC reveals an inverse correlation between the levels of AC and their response to exogenous C-6-ceramide. Furthermore, over-expression of AC in SCC-1 cells increased resistance to Fas induced cell killing. Conversely, down-regulation of AC using specific AC small interfering RNA (siRNA) sensitized the SCC-1 cancer cell line to Fas-induced apoptosis. Finally, we show that the AC inhibitor LCL 204 can sensitize HNSCC cell lines to Fas-induced apoptosis both in vitro and in a xenograft model in vivo, suggesting that the combination of FasL gene therapy and LCL 204 may become a new treatment option for advanced-stage head and neck cancer. PMID- 17426711 TI - An efficient and safe herpes simplex virus type 1 amplicon vector for transcriptionally targeted therapy of human hepatocellular carcinomas. AB - Our previous studies have shown that transgene expression could be targeted to proliferating cells when cell cycle transcriptional regulatory elements were incorporated into herpes simplex virus type 1 (HSV-1) amplicon backbone vectors. In the study reported here, we further demonstrated the transcriptional activation of transgene expression in association with the onset of cellular proliferation using the mouse partial hepatectomy model. Moreover, transcriptional regulation could be rendered specific to human hepatocellular carcinoma (HCC) cells by inserting the chimeric gene Gal4/NF-YA under the regulation of the HCC-specific hybrid promoter. The hybrid promoter, which consists of four copies of the apolipoprotein E (ApoE) enhancer element inserted upstream of the human alpha1-antitrypsin(hAAT) promoter, induced an higher level of transcription than other liver-specific promoters such as alpha-fetoprotein (AFP) and albumin (Alb) promoter. As a consequence, the enhancement of tissue specific expression in the context of Gal4/NF-YA fusion proteins enabled the monitoring of transgene expression using a bioluminescence imaging system. Furthermore, these vectors have been shown to be non-toxic and exhibited potent infectivity for proliferating primary HCC cells and HCC cell lines. Together, these results demonstrated that the new hybrid vectors could provide options for the design of safe and efficient systemic gene therapeutic strategies for human HCC. PMID- 17426712 TI - Hsp40 molecules that target to the ubiquitin-proteasome system decrease inclusion formation in models of polyglutamine disease. AB - We studied the ability of heat shock, DnaJ-like-1 (HSJ1) proteins (which contain DnaJ and ubiquitin-interacting motifs) to reduce polyglutamine-mediated inclusion formation. The experiments demonstrated that expression of heat shock protein 70 (hsp70), hsp40, HSJ1a, and HSJ1b significantly reduced protein inclusion formation in a model of spinal and bulbar muscular atrophy (SBMA). HSJ1a also mediated a significant decrease in the number of inclusions formed in a primary neuronal model of protein aggregation. Studies to elucidate the mechanisms underlying these reductions showed that hsp70 and hsp40 increased chaperone mediated refolding. In contrast, expression of HSJ1 proteins did not promote chaperone activity but caused an increase in ubiquitylation. Furthermore, HSJ1a was associated with a ubiquitylated luciferase complex, and in the presence of HSJ1a but not an HSJ1a UIM mutant (HSJ1a-deltaUIM) there was a reduction in luciferase protein levels. Together these results show that HSJ1 proteins mediated an increase in target protein degradation via the ubiquitin-proteasome system (UPS). We also found that the expression of HSJ1a significantly decreased the number of neurons containing inclusions in an in vivo model of polyglutamine disease. These findings indicate that targeted modification of the UPS to facilitate degradation of misfolded proteins may represent a highly effective therapeutic avenue for the treatment of polyglutamine disease. PMID- 17426713 TI - Sustained AAV-mediated dystrophin expression in a canine model of Duchenne muscular dystrophy with a brief course of immunosuppression. AB - Adeno-associated virus-based vector (AAV)-mediated gene delivery has been successful in some animal models of human disease such as the mdx mouse model of human Duchenne muscular dystrophy (DMD). However, recent evidence of immune mediated loss of vector persistence in dogs and humans suggests that immune modulation might be necessary to achieve successful long-term transgene expression in these species. We have previously demonstrated that direct intramuscular injection of AAV2 or AAV6 in wild-type random-bred dogs resulted in a robust immune response to capsid or capsid-associated proteins. We now demonstrate that a brief course of immunosuppression with a combination of anti thymocyte globulin (ATG), cyclosporine (CSP), and mycophenolate mofetil (MMF) is sufficient to permit long-term and robust expression of a canine micro-dystrophin (c-micro-dys) transgene in the skeletal muscle of a dog model for DMD (canine X linked muscular dystrophy, or cxmd dog) and that its expression restored localization of components of the dystrophin-associated protein complex at the muscle membrane. This protocol has potential applications to human clinical trials to enhance AAV-mediated therapies. PMID- 17426714 TI - Does prolonged clopidogrel therapy improve outcome in patients with drug-eluting or bare-metal stents? PMID- 17426715 TI - Rosiglitazone monotherapy for type 2 diabetes mellitus-too soon to ADOPT? PMID- 17426716 TI - Addressing the big challenge of obesity: how to prevent regain of lost weight. PMID- 17426717 TI - Intensive versus moderate blood-pressure control in normotensive patients with type 2 diabetes. PMID- 17426718 TI - Taking the weight off the fistula: the importance of vein transposition and fistula evaluation in obese patients. PMID- 17426719 TI - Clinical correlates of Lewy-body-related disorders: splitting and lumping or lumping and splitting? PMID- 17426720 TI - Sunitinib--a new approach following failure of imatinib in patients with advanced gastrointestinal stromal tumor. PMID- 17426721 TI - Does ofloxacin protect against BCG-related toxic effects in patients with bladder cancer? PMID- 17426722 TI - Do dehydroepiandrosterone supplements have anti-aging potential? PMID- 17426723 TI - Abundance of the POLG disease mutations in Europe, Australia, New Zealand, and the United States explained by single ancient European founders. AB - We reported previously that the DNA polymerase gamma (POLG) W748S mutation, a common cause of mitochondrial recessive ataxia syndrome (MIRAS), has a common ancient founder for all the disease chromosomes in Finland, Norway, United Kingdom, and Belgium. Here, we present results showing that the same ancestral chromosome underlies MIRAS and Alpers syndrome in Australia and New Zealand. Furthermore, we show that a second common POLG mutation, A467T, also shows common European ancestry: patients from Australia, New Zealand, and the United States share a common haplotype with the previously reported European patients. These data of ancestral haplotypes indicate that the POLG locus is quite stable and that the recessive W748S and A467T mutations, and probably also G848S, have occurred once in history. They have effectively spread to populations of European descent with carrier frequencies up to 1% in several populations. Our data predict that these mutations are common causes of ataxia and Alpers disease in the Western world. PMID- 17426724 TI - Evidence for gene x smoking exposure interactions in a genome-wide linkage screen of asthma and bronchial hyper-responsiveness in EGEA families. AB - Asthma and bronchial hyper-responsiveness (BHR), an asthma-related phenotype, result from many genetic (G) and environmental (E) factors. Passive exposure to tobacco smoke (ETS) in early life is one of these risk factors. Following a genome scan for asthma and associated phenotypes conducted in 295 French Epidemiological study on the Genetics and Environment of Asthma, our present aim was to investigate interactions between genetic susceptibility to asthma and to BHR with passive ETS using two different methods: the predivided sample test (PST) and the mean interaction test (MIT). PST and MIT consider the identical by descent (identity by descent) distribution at all markers in affected sib-pairs with 2, 1 or 0 sib(s) exposed to ETS. While the PST allows detection of both linkage and G x E interaction, the MIT tests for linkage by taking into account a possible interaction. Among the six regions detected at P10% of total calories, in only 6% was fibre intake > or =20 g/1000 kcal (considered ideal), and in 25% it was > or =15 g/1000 kcal (acceptable). CONCLUSIONS: These results indicate that compliance to dietary recommendations is not completely satisfactory, even in Italy. Calorie intake is a bit elevated, given the high BMI of our diabetic population. As to dietary composition, there are two crucial issues: the high intake of saturated fat and--most importantly--the low intake of fibre. All strategies aiming to a proper implementation of guidelines should take these results into due account. PMID- 17426740 TI - Vitamin C and exercise-induced immunodepression. PMID- 17426741 TI - Consumption of cruciferous vegetables and glucosinolates in a Spanish adult population. AB - OBJECTIVE: To assess the intake of glucosinolates and cruciferous vegetables among Spanish adults. DESIGN: Cross-sectional analysis of a prospective cohort study. SETTING: The Spanish cohort of the European Prospective Investigation into Cancer and Nutrition (EPIC). SUBJECTS: We analysed data from 40 684 men and women aged 35-64 years from the EPIC-Spain cohort. The usual diet was assessed by means of the dietary history method, and glucosinolate intake was calculated using a published food composition database. RESULTS: The average intake of cruciferous vegetables was 11.3 g/day, accounting for about 5% of total vegetable consumption, whereas the daily intake of total glucosinolates was 6.5 mg, among which 35% were of indole type. The absolute intake of glucosinolates was in average higher in men than in women (6.8 vs 6.2 mg/day), whereas glucosinolate density per energy unit was higher in women's diet (3.4 vs 2.7 mg/4200 kJ). Northern regions consumed in average 36% more glucosinolates than Southern regions (7.3 vs 5.4 mg/day). There was a positive association of glucosinolate intake with body mass index, physical activity, educational level and an inverse relationship with alcohol consumption. CONCLUSIONS: Contrary to the pattern seen for total vegetable intake, our estimate of consumption of cruciferous vegetables, and hence of glucosinolates, is relatively low within Europe, which in turn is lower than in North America and several Asian populations. PMID- 17426742 TI - Muesli with 4 g oat beta-glucans lowers glucose and insulin responses after a bread meal in healthy subjects. AB - OBJECTIVE: To evaluate the impact of an extruded muesli product based on beta glucan-rich oat bran on postprandial glycaemia and insulinaemia. SUBJECT/DESIGN: The study is divided in two series. Blood glucose and serum insulin responses were studied after subjects consuming test meals including a serving of muesli with 3 g (series 1) and 4 g (series 2) of beta-glucans, respectively. The muesli was a component in a single serving packet with muesli and yoghurt. This was served together with white wheat bread in the morning after an overnight fast. The compositions were standardized to contain 50 g available carbohydrates. As a reference meal a serving packet without beta-glucans was included. The study was performed at Applied Nutrition and Food Chemistry, Lund University, Sweden. Nineteen and thirteen healthy volunteers with normal body mass index were recruited for series 1 and 2, respectively. RESULTS: Muesli with 3 g of beta glucans, included in a mixed bread meal, gave no significant differences in glycaemic response compared to a reference meal without muesli and beta-glucans. In contrast, muesli with 4 g of beta-glucans significantly (P<0.05) lowered the glucose and insulin responses compared to the reference meal. CONCLUSIONS: Muesli enriched with 4 g of beta-glucans reduces postprandial glucose and insulin levels to a breakfast based on high glycaemic index products. A total of 4 g of beta glucans from oats seems to be a critical level for a significant decrease in glucose and insulin responses in healthy people. PMID- 17426743 TI - The impact of freezing and toasting on the glycaemic response of white bread. AB - OBJECTIVE: To investigate the impact of freezing and toasting on the glycaemic response of white bread. SUBJECTS/METHODS: Ten healthy subjects (three male, seven female), aged 22-59 years, recruited from Oxford Brookes University and the local community. A homemade white bread and a commercial white bread were administered following four different storage and preparation conditions: (1) fresh; (2) frozen and defrosted; (3) toasted; (4) toasted following freezing and defrosting. They were administered randomized repeated measures design. Incremental blood glucose, peak glucose response, 2 h incremental area under the glucose response curve (IAUC). RESULTS: The different storage and preparation conditions resulted in lower blood glucose IAUC values compared to both types of fresh white bread. In particular, compared to the fresh homemade bread (IAUC 259 mmol min/l), IAUC was significantly lower when the bread was frozen and defrosted (179 mmol min/l, P<0.05), toasted (193 mmol min/l, P<0.01) and toasted following freezing and defrosting (157 mmol min/l, P<0.01). Similarly, compared to the fresh commercial white bread (253 mmol min/l), IAUC was significantly lower when the bread was toasted (183 mmol min/l, P<0.01) and frozen, defrosted and toasted (187 mmol min/l, P<0.01). CONCLUSIONS: All three procedures investigated, freezing and defrosting, toasting from fresh, and toasting following freezing and defrosting, favourably altered the glucose response of the breads. This is the first study known to the authors to show reductions in glycaemic response as a result of changes in storage conditions and the preparation of white bread before consumption. In addition, the study highlights a need to define and maintain storage conditions of white bread if used as a reference food in the determination of the glycaemic index of foods. PMID- 17426744 TI - Urinary excretion of 13 dietary flavonoids and phenolic acids in free-living healthy subjects - variability and possible use as biomarkers of polyphenol intake. AB - OBJECTIVE: Estimation of dietary intake of polyphenols is difficult, due to limited availability of food composition data and bias inherent to dietary assessment methods. The aim of the present study was to evaluate whether we could detect polyphenols and their metabolites in a spot urine sample in a free-living human population and whether it was related to those observed in 24-h urine samples, for potential use as a biomarkers of polyphenol intake. SUBJECTS: Four 24-h urine samples and two spot urine samples were collected from 154 participants of the SU.VI.MAX cohort (a randomized primary-prevention trial evaluating the effect of daily antioxidant supplementation on chronic diseases) in two separate studies over, respectively, a 7- and 2-day periods. Thirteen polyphenols and metabolites (chlorogenic acid (CGA), caffeic acid (CA), m coumaric acid (mCOU), gallic acid (GA), 4-O-methylgallic acid (MeGA), quercetin (Q), isorhamnetin (MeQ), kaempferol (K), hesperetin (HESP), naringenin (NAR), phloretin (PHLOR), enterolactone (ENL) and enterodiol (END) were measured using HPLC-ESI-MS-MS. RESULTS: Correlations between the urinary excretion levels were observed. The most significant were explained by metabolic filiations (CGA/CA, CA/mCOU, GA/MeGA, Q/MeQ, NAR/PHLOR, ENL/END) or co-occurrence in a same food source (NAR/HESP). Concentrations in spot samples correlated with those in 24-h urine sample (P<0.02, except for CA and for MeQ). Intra-individual variations were smaller than inter-individual variations for all polyphenols (P<0.01) except for MeGA and for PHLOR. CONCLUSION: These results show that these polyphenols and metabolites are useful biomarkers for polyphenol intake. PMID- 17426745 TI - Identifying sources of reporting error using measured food intake. AB - OBJECTIVE: To investigate the magnitude and relative contribution of different sources of measurement errors present in the estimation of food intake via the 24 h recall technique. DESIGN: We applied variance decomposition methods to the difference between data obtained from the USDA's Automated Multiple Pass Method (AMPM) 24-h recall technique and measured food intake (MFI) from a 16-week cafeteria-style feeding study. The average and the variance of biases, defined as the difference between AMPM and MFI, were analyzed by macronutrient content, subject and nine categories of foods. SUBJECTS: Twelve healthy, lean men (age, 39+/-9 year; weight, 79.9+/-8.3 kg; and BMI, 24.1+/-1.4 kg/m2). RESULTS: Mean food intakes for AMPM and MFI were not significantly different (no overall bias), but within-subject differences for energy (EI), protein, fat and carbohydrate intakes were 14, 18, 23 and 15% of daily intake, respectively. Mass (incorrect portion size) and deletion (subject did not report foods eaten) errors were each responsible for about one-third of the total error. Vegetables constituted 8% of EI but represented >25% of the error across macronutrients, whereas grains that contributed 32% of EI contributed only 12% of the error across macronutrients. CONCLUSIONS: Although the major sources of reporting error were mass and deletion errors, individual subjects differed widely in the magnitude and types of errors they made. PMID- 17426746 TI - How do women change osteoporosis-preventive behaviours in their children? AB - OBJECTIVE: We have reported that a lifestyle intervention with mothers improved calcium intake and physical activity in both mothers and their children. In this study, we aimed to describe the strategies and approaches used by these mothers to improve their children's calcium intake and physical activity. DESIGN: A qualitative study using semistructured interviews. SETTING: Population-based convenience sample. SUBJECTS: A subsample of 39 mothers were taken from a population-based random sample of 354 mothers who had participated in the original osteoporosis-prevention trial. RESULTS: Mothers described specific dietary changes they made to increase their children's calcium intake. They also described strategies for improving calcium intake and physical activity such as raising awareness of the importance of calcium; ensuring calcium-rich foods were accessible; assessing their children's likes and dislikes and working within these; role modelling; information provision; taking a balanced approach to attempting behaviour change; and encouraging activities that they could do with their children. Mothers emphasized the general importance of a balanced diet and lifestyle, rather than just focussing on lifestyle factors specific to osteoporosis prevention. CONCLUSION: Even without specific guidance, mothers are adept at developing strategies to apply to changing lifestyle behaviours in their children and identifying barriers to change. These results provide information, which could be incorporated into future interventions for lifestyle change in children and also provide further support for considering parent-focused approaches to this problem. PMID- 17426747 TI - Determination of the glycaemic index of various staple carbohydrate-rich foods in the UK diet. AB - OBJECTIVE: To determine the glycaemic index (GI) of various staple carbohydrate rich foods in the UK diet, and to consider the factors influencing the GI of foods. DESIGN: Subjects were served with 25 or 50 g portions of glucose on three occasions, followed by a selection of test foods providing an equal amount of available carbohydrate, in random order. Each test food was consumed by 10 subjects. Capillary blood glucose levels were measured in the fasted state and over the 120 min following commencement of consumption of the foods. SETTING: The study was carried out in a research institute (MRC Human Nutrition Research, Cambridge, UK). SUBJECTS: Forty-two healthy adult volunteers were studied. METHODS: The GI values of 33 foods were measured according to the WHO/FAO recommended methodology. These foods included various breads, breakfast cereals, pasta, rice and potatoes, all of which were commercially available in the UK. CONCLUSIONS: The results illustrate a number of factors which are important in influencing the GI of a food, highlighting the importance of measuring the GI of a food, rather than assuming a previously published value for a similar food. This is useful both to researchers analysing dietary surveys or planning intervention studies, and also to health professionals advising individuals on their diets. PMID- 17426748 TI - Divergent in vitro and in vivo lipid peroxidation in the postprandial phase of patients with type I diabetes mellitus. AB - OBJECTIVE: The two- to fourfold higher risk of cardiovascular disease in diabetes mellitus is more strongly predicted by the postprandial than by the fasting blood glucose and lipids. We aimed to investigate the impact of postprandial changes in serum lipoprotein fractions on lipid peroxidation in type I diabetes mellitus (T1DM). DESIGN: This was a prospective observational study. SETTING: The study was performed at Antwerp University Hospital, Belgium. SUBJECTS: Twenty-three well-controlled T1DM patients were included. INTERVENTION: Patients received a standard breakfast and lunch (>50% energy as fat). Blood was sampled at fasting (F), after the post-breakfast hyperglycemic peak (BP), just before lunch (B), after the post-lunch hyperglycemic peak (LP), after the post-lunch dale (LD) and 5 h after lunch (L) for the measurement of serum lipids, lipoprotein subfraction composition, alpha-tocopherol and lipid peroxidation in vivo and in vitro. RESULTS: Serum triacylglycerols (Tgs) increased (from 1.03+/-0.40 at F to 1.60+/ 0.87 mmol/l at LP, P=0.001), but cholesterol decreased by 12% in parallel with alpha-tocopherol (from 4.43+/-0.76 at F to 4.12+/-0.82 micromol/mmol total lipid at B, P=0.006). Although plasma malondialdehyde increased from 1.02+/-0.36 at F to 1.14+/-0.40 micromol/L at LP, P=0.03, copper-induced in vitro peroxidation decreased in the low-density lipoprotein and high-density lipoprotein fractions. CONCLUSIONS: In well-controlled T1DM patients moderate postprandial increases in serum Tgs are accompanied by a relative deficiency in alpha-tocopherol. Lipid peroxidation in vivo increases but cannot be ascribed to changes in the susceptibility of lipoproteins to copper-induced in vitro peroxidation. PMID- 17426749 TI - Genetic susceptibility to heart disease in Canada: lessons from patients with familial hypercholesterolemia. AB - Much of the recent progress in treating patients with heart disease due to narrowed coronary arteries has resulted from studying disease evolution in patients with rare monogenic forms of disease. For instance, autosomal dominant familial hypercholesterolemia (FH, MIM (Mendelian Inheritance in Man) 143890) typically results from heterozygous mutations in LDLR encoding the low-density lipoprotein (LDL) receptor. Deficient LDLR activity results in elevated circulating LDL cholesterol, which accumulates within blood vessel walls, forming arterial plaques that can grow and eventually occlude the arterial lumen. Heterozygous LDLR mutations are usually detected using exon-by-exon sequence analysis (EBESA) of genomic DNA, a technology that has identified approximately 50 mutations in heterozygous FH (HeFH) subjects in Ontario. However, approximately 35% of Ontario HeFH patients had no EBESA-identified LDLR mutation. The diagnostic gap relates both to the genetic heterogeneity of FH and also to inadequate sensitivity of EBESA to detect certain mutation types, such as large deletions or insertions in LDLR. By means of a dedicated method to detect copy number variations (CNVs), additional heterozygous mutations in LDLR ranging from approximately 500 to >15 000 bases were uncovered, accounting for most of the remainder of Ontario HeFH patients. The appreciation of the key role of genomic CNVs in disease coincides with recent genome-wide mapping studies demonstrating that CNVs are common in apparently healthy people. CNVs thus represent a new level of genomic variation that is both an important mechanism of monogenic disease and a contributor to genomic variation in the general population; as well, it may have implications for evolution, biology, and possibly susceptibility to common complex diseases. PMID- 17426750 TI - Phenetic relationships among different races of Striga gesnerioides (Willd.) Vatke from West Africa. AB - Striga gesnerioides is a root hemiparasite that primarily parasitizes dicotyledonous species, including cowpea (Vigna unguiculata L.) and other legumes. Based on the differential resistance response of various cultivars, landraces, and breeding lines, it has been proposed that several distinct races of cowpea-parasitic S. gesnerioides exist in West Africa. In this study, we used amplified fragment length polymorphism profile analysis to examine the genetic variability within and among populations of cowpea-parasitic S. gesnerioides within the suspected distribution range of a particular race, and statistical clustering methods to define the phenetic relationships of the various races in West Africa. Our data indicate that genetic variability within and among populations of each of the previously recognized races of cowpea-parasitic S. gesnerioides is extremely low. On the basis of genotypic profile and host differential resistance responses, 2 previously unknown races were identified. Of the 7 races now identifiable, races SG1 (from Burkina Faso) and SG5 (from Cameroon) are the most closely related, and SG4 (from Benin) and SG3 (from Niger/Nigeria) are the most divergent. SG6, a new race of the parasite identified in Senegal, was found to be the most genetically similar to SG4 from Benin. We also demonstrate that a hypervirulent isolate of the S. gesnerioides from Zakpota, in the Republic of Benin, is genotypically distinct from other populations of SG4, thereby warranting designation as a separate race, which we called SG4z. To further support our race classification scheme, we identified a group of molecular markers that effectively discriminate each of the various races. Finally, we show that an isolate (designated SG4i) of the wild legume Indigofera hirsuta--parasitic S. gesnerioides is genetically distinct and significantly diverged from the various races of cowpea-parasitic S. gesnerioides. Our data suggest that both geographic isolation and host-driven selection are critical factors defining race formation in S. gesnerioides in West Africa. PMID- 17426751 TI - Length, orientation, and plant host influence the mutation frequency in microsatellites. AB - Microsatellites are simple, tandem DNA repeats that represent unstable regions of the genome. They undergo frequent changes in tract length by base additions or deletions due to DNA polymerase slippage during replication. To characterize factors affecting the frequency of spontaneous mutations occurring in microsatellites in plants, a reporter system was used in Arabidopsis thaliana and tomato (Lycopersicon esculentum). The beta-glucuronidase (GUS) reporter system was used to measure the mutation frequency in various microsatellites (G(7), G(10), G(13), G(16), and C(16)) in somatic tissues. Our results indicate that this frequency increases with the number of repeats: a G(16) tract was almost 80 fold more mutable than a G(7) tract. Furthermore, the frequency of mutations depends on repeat orientation, as G(16) was 3-fold more mutable than C(16). The mutation rate was also found to differ markedly in Arabidopsis and tomato for an identical microsatellite. Indeed, Arabidopsis showed a 5-fold higher mutation frequency than tomato with the same G(7) reporter construct. Finally, mutation in a G(16) tract was frequent enough that mutations transmitted germinally to the next generation could be detected at a relatively high frequency. PMID- 17426752 TI - Chiasmata and chromosome breakages are related to crossing over in Drosophila ananassae males. AB - A cytogenetic analysis of male crossing over in Drosophila ananassae revealed that cytological exchanges resulted in genetic crossing over, and that chiasma frequency and the genetic recombination correlated positively in chromosomes 2 and 3. Furthermore, the frequency of chromosome breakages correlated positively with chiasma frequency. Paracentric inversion heterozygosity had no detectable influence on the chromosome pairing or exchange events within the inversion loop at meiosis. Scoring of the chiasma demonstrated that males homozygous for the previously mapped enhancers of male crossing over had low frequencies of chiasmata, whereas higher frequencies of chiasmata were observed in males heterozygous for enhancers. The results presented here indicate that the genetic factors controlling male crossing over are involved in the origin of chromosome breakages and in exchange events. PMID- 17426753 TI - Polytene chromosomes and phylogenetic relationships of Chironomus atrella (Diptera: Chironomidae) in North America. AB - The identity of Chironomus atrella Townes has been confusing because the name has been used for at least 2 quite different species. This situation is clarified karyosystematically by describing the banding patterns and chromosomal polymorphisms from a number of locations in Canada and the US. Most populations show only moderate levels of polymorphism (average heterozygosity, 0.36), although in some samples from shallow waters, the level of polymorphism is much higher (average heterozygosity, up to 0.92). The banding patterns of the polytene chromosomes are either identical or closely related to those found in Holarctic species with a northern distribution. These patterns and the distribution of inversions in the C. atrella populations are consistent with a progenitor that colonized North America across the Bering Strait and spread down the Rocky Mountain chain; at the same time, new gene combinations developed that allowed it to spread eastward over the majority of the continent. PMID- 17426754 TI - Construction and characterization of the first bacterial artificial chromosome library for the cotton species Gossypium barbadense L. AB - As the second most widely cultivated cotton, Gossypium barbadense is well known for its superior fiber properties and its high levels of resistance to Fusarium and Verticillium wilts. To enhance our ability to exploit these properties in breeding programs, we constructed the first bacterial artificial chromosome (BAC) library for this species. The library contains 167 424 clones (49 920 BamHI and 117 504 HindIII clones), with an estimated average insert size of 130 kb. About 94.0% of the clones had inserts over 100 kb, and the empty clones accounted for less than 4.0%. Contamination of the library with chloroplast clones was very low (0.2%). Screening the library with locus-specific probes showed that BAC clones represent 6.5-fold genome equivalents. This high-quality library provides an additional asset with which to exploit genetic variation for cotton improvement. PMID- 17426755 TI - A sequence related to rice Pong transposable element displays transcriptional activation by in vitro culture and reveals somaclonal variations in maize. AB - Miniature inverted-repeat transposable elements (MITEs) are nonautonomous elements that are abundant in plant genomes. The rice MITE mPing was shown to be mobilized by anther culture, and the associated transposon Pong was shown to transpose actively in an Oryza sativa 'indica' rice cell-culture line. We have identified 3 sequences in maize named ZmTPAPong-like 1, 2, and 3 that displayed homology with the transposase of Pong. Here, we show that these sequences are differentially expressed during the in vitro androgenetic process in maize. We also demonstrate that the ZmTPAPong-like 1 and 3 sequences reveal somaclonal variations among plants regenerated from the calli of a doubled haploid line. These data suggest that the ZmTPAPong-like sequences could form part of a Zea mays element related to the rice Pong element. The possible activation of this newly discovered element under stress conditions is discussed. PMID- 17426756 TI - Population structure of Sclerotinia sclerotiorum in an Australian canola field at flowering and stem-infection stages of the disease cycle. AB - Populations of the ascomycete pathogen Sclerotinia sclerotiorum sampled from a canola field were analysed using microsatellite markers. Fifty isolates were collected from ascospore-infested canola petals and, later in the season, another 55 isolates were obtained from stem lesions; these isolates were used to compare inoculum and disease-causing populations. Fifty-five unique haplotypes were identified, with gene diversity ranging from 0.40 to 0.71. Genotypic diversity was higher in the inoculum population than it had been in the previous year, but analysis of molecular variance (AMOVA) showed that less than 10% of the variation was attributable to differences between the 2 years. Genotypic disequilibrium measures were consistent with the occurrence of both clonal reproduction and out crossing. There was no significant population subdivision between the ascospore and stem-lesion populations, as measured with fixation indices (R(ST) = 0.015, p = 0.90) and AMOVA, suggesting that there are no genetically defined subgroups of isolates more likely to proceed from petal colonization to cause stem infection. This might be because S. sclerotiorum possesses wide-ranging pathogenicity mechanisms that account for the lack of host specificity observed to date. PMID- 17426757 TI - A novel strategy for identifying mutations that sensitize Drosophila eye development to caffeine and hydroxyurea. AB - This report describes a novel strategy for isolating Drosophila mutants with conditional eye phenotypes that should be generally applicable for identifying genes required for cellular responses to specific drugs. To test the strategy, we screened 3 of the 5 major chromosome arms for hydroxyurea- and (or) caffeine sensitive (huc) mutants, and isolated mutations affecting 5 different complementation groups. Most of these were represented by single alleles; however, we also isolated multiple alleles of huc(29DE) gene, an essential gene that is also associated with a nonconditional pupal lethal phenotype. We also identified huc(95E) mutants, which are extremely sensitive to caffeine. Although huc(95E) is a nonessential gene, mutant imaginal disc cells undergo caffeine dependent apoptosis, and huc(95E) gene function is required for the viability of the organism when mutant larvae are exposed to levels of caffeine that controls can easily tolerate. We have mapped the cytological positions of huc(29D) and huc(95E) as a first step toward molecularly characterizing the relevant genes. PMID- 17426758 TI - Environmental stress and life-stage dependence on the detection of heterozygosity fitness correlations in the European eel, Anguilla anguilla. AB - Heterozygosity-fitness correlations (HFCs) have been reported in populations of many species, although HFCs can clearly vary across species, conspecific populations, temporal samples, and sexes. We studied (i) the temporal stability of the association between genetic variation and growth rate (length and mass increase) and (ii) the influence of genetic variability on survival in the European eel (Anguilla anguilla L). HFCs were assessed using genotypes from 10 allozyme and 6 microsatellite markers in 22-month-old experimental individuals. The results were compared with those of a previous study carried out in 12-month old individuals, in which more heterozygous individuals showed a significantly faster growth rate. In contrast, 22-month-old individuals showed no evidence that genetic variability was correlated with growth rate. Additionally, heterozygous individuals did not show a higher survival rate compared with more homozygous individuals after either handling stress or parasite infection. The decrease in HFCs over time is consistent with the general prediction that differences in growth and survival among individuals are maximal early in life and in our case most likely due to the relaxation of environmental conditions related to population-density effects. Alternatively, the decline in HFCs could be attributed to either ontogenetic variance in gene activity between 12- and 22 month-old individuals or differential mortality leaving only the largest individuals. PMID- 17426759 TI - Defeating numts: semi-pure mitochondrial DNA from eggs and simple purification methods for field-collected wildlife tissues. AB - Mitochondrial DNA (mtDNA) continues to play a pivotal role in phylogeographic, phylogenetic, and population genetic studies. PCR amplification with mitochondrial primers often yields ambiguous sequences, in part because of the co amplification of nuclear copies of mitochondrial genes (numts) and true mitochondrial heteroplasmy arising from mutations, hybridization with paternal leakage, gene duplications, and recombination. Failing to detect numts or to distinguish the origin of such homologous sequences results in the incorrect interpretation of data. However, few studies obtain purified mtDNA to confirm the mitochondrial origin of the first reference sequences for a species. Here, we demonstrate the importance and ease of obtaining semi-pure mtDNA from wildlife tissues, preserved under various typical field conditions, and investigate the success of 3 commercial extraction kits, cesium-chloride gradient mtDNA purification, long-template PCR amplification, cloning, and more species-specific degenerate primers. Using more detailed avian examples, we illustrate that unfertilized or undeveloped eggs provide the purest sources of mtDNA; that kits provide an alternative to cesium-chloride gradient methods; and that long template PCR, cloning, and degenerate primers cannot be used to produce reliable mitochondrial reference sequences, but can be powerful tools when used in conjunction with purified mtDNA stocks to distinguish numts from true heteroplasmy. PMID- 17426760 TI - Three new satellite sequences and a mobile element found inside HSP70 introns of the Mediterranean mussel (Mytilus galloprovincialis). AB - We report the characterization of 3 new repetitive sequences from the bivalve mollusc Mytilus galloprovincialis, designated Mg1, Mg2, and Mg3, with monomer lengths of 169, 260, and 70 bp, respectively. The 3 repeats together constitute approximately 7.8% of the M. galloprovincialis genome and were found, together with ApaI-type 2 repeats, inside the introns of 2 genes of the HSP70 family, hsc70 and hsc71. Both the monomer length and the genomic content of the repeats indicate satellite sequences. The Mg1 repetitive region and its flanking sequences exhibit significant homology to CvE, a member of the Pearl family of mobile elements found in the eastern oyster (Crassostrea virginica). Thus, the whole homologous region is designated MgE, the first putative transposable element characterized in M. galloprovincialis. The ApaI, Mg2, and Mg3 repeats are continuously arranged inside the introns of both the hsc70 and hsc71 genes. The presence of perfect inverted repeats flanking the ApaI-Mg2-Mg3 repetitive region, as well as a sequence analysis of the repeats, indicates a transposition-like insertion of this region. The genes of the HSP70 family are highly conserved, and the presence of repetitive DNA or of mobile elements inside their introns is reported here for the first time. PMID- 17426761 TI - Genetic diversity and geographical dispersal in grapevine clones revealed by microsatellite markers. AB - Intravarietal genetic diversification associated with geographical dispersal of a vegetatively propagated species was studied using grapevine Vitis vinifera L. 'Cabernet Sauvignon' as a model. Fifty-nine clonal samples obtained from 7 countries (France, Chile, Spain, Australia, Hungary, USA, and Italy) were analyzed using 84 microsatellite markers. Eighteen polymorphic microsatellite loci (21.4%) were detected, finding 22 different genotypes in the population analyzed with a genetic similarity of over 97%. The presence of chimeric clones was evidenced at locus VMC5g7 by means of a segregation analysis of descendants by self-pollination of a triallelic Chilean clone and by somatic embryogenesis analysis, showing a mutation in L2 cell layer. Only 2 clones (obtained from France and Australia) presented the ancestral genotype, and the most divergent genotype was exhibited by another French clone, which had accumulated 5 somatic mutations. The 2 largest populations considered (from France and Chile) showed a clear divergency in the polymorphisms detected. These antecedents enabled the tracing of geographical dispersal with a phylogenetic hypothesis supporting France as the center of origin of diversification of Cabernet Sauvignon. The results obtained could help to explain diversification processes in other grapevine cultivars. The possibility that this kind of genetic variability occurs in other vegetatively propagated species is discussed, focusing on possible fingerprinting applications. PMID- 17426762 TI - Nucleotide-binding site (NBS) profiling of genetic diversity in durum wheat. AB - Molecular markers are effective tools to investigate genetic diversity for resistance to pathogens. NBS (nucleotide-binding site) profiling is a PCR (polymerase chain reaction)-based approach to studying genetic variability that specifically targets chromosome regions containing R-genes and R-gene analogues. We used NBS profiling to measure genetic diversity among 58 accessions of durum wheat. Mean polymorphism rates detected using MseI and AluI as restriction enzymes were 34% and 22%, respectively. Mean number of polymorphisms per enzyme primer combination was equal to 23.8 +/- 5.9, ranging from 13 to 31 polymorphic bands. In total, 96 markers over 190 indicated a good capacity to discriminate between accessions (the polymorphic index content ranging from 0.30 to 0.50). The results obtained with NBS profiling were compared with simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) data of the same set of accessions. The genetic distances computed with 190 NBS profiling markers were in close agreement with those obtained with AFLP and SSR markers (r = 0.73 and 0.76, respectively). Our results indicate that NBS profiling provides an effective means to investigate genetic diversity in durum wheat. PMID- 17426763 TI - Isolation and characterization of the pea cytochrome c oxidase Vb gene. AB - Three copies of the gene that encodes cytochrome c oxidase subunit Vb were isolated from the pea (PscoxVb-1, PscoxVb-2, and PscoxVb-3). Northern Blot and reverse transcriptase-PCR analyses suggest that all 3 genes are transcribed in the pea. Each pea coxVb gene has an N-terminal extended sequence that can encode a mitochondrial targeting signal, called a presequence. The localization of green fluorescent proteins fused with the presequence strongly suggests the targeting of pea COXVb proteins to mitochondria. Each pea coxVb gene has 5 intron sites within the coding region. These are similar to Arabidopsis and rice, although the intron lengths vary greatly. A phylogenetic analysis of coxVb suggests the occurrence of gene duplication events during angiosperm evolution. In particular, 2 duplication events might have occurred in legumes, grasses, and Solanaceae. A comparison of amino acid sequences in COXVb or its counterpart shows the conservation of several amino acids within a zinc finger motif. Interestingly, a homology search analysis showed that bacterial protein COG4391 and a mitochondrial complex I 13 kDa subunit also have similar amino acid compositions around this motif. Such similarity might reflect evolutionary relationships among the 3 proteins. PMID- 17426764 TI - Tracing B-genome chromatin in Brassica napus x B. juncea interspecific progeny. AB - We used polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques to demonstrate the presence of Brassica B-genome chromosomes and putative B-genome introgressions in B. napus x B. juncea interspecific progeny. The B-genome--specific repeat sequence pBNBH35 was used to generate PCR products and FISH probes. The highest frequencies of viable progeny were obtained when B. napus was the maternal parent of the interspecific hybrid and the first backcross. B-genome--positive PCR assays were found in 34/51 fertile F2 progeny (67%), which was more than double the proportion found in fertile BC(1) progeny. Four B-genome--positive F(2)-derived families and 1 BC(1)-derived family were fixed or segregating for B. juncea morphology in the F(4) and BC(1)S(2), respectively, but in only 2 of these families did B. juncea-type plants exhibit B. juncea chromosome count (2n = 36) and typical B-genome FISH signals on 16 chromosomes. The remaining B. juncea-type plants had B. napus chromosome count (2n = 38) and no B-genome FISH signals, except for 1 exceptional F(4)-derived line that exhibited isolated and weak B-genome FISH signals on 11 chromosomes and typical A-genome FISH signals. B. juncea morphology was associated with B-genome- positive PCR signals but not necessarily with 16 intact B-genome chromosomes as detected by FISH. B-genome chromosomes tend to be eliminated during selfing or backcrossing after crossing B. juncea with B. napus, and selection of lines containing B-genome chromatin during early generations would be promoted by use of this B-genome repetitive marker. PMID- 17426765 TI - Colocalization of a partially dominant gene for yellow seed colour with a major QTL influencing acid detergent fibre (ADF) content in different crosses of oilseed rape (Brassica napus). AB - Quantitative trait loci (QTLs) contributing to yellow seed colour and acid detergent fibre (ADF) were localized and compared in 3 mapping populations developed from 2 crosses (designated 'YE1' and 'YE2') between 2 distinct sources of true-breeding yellow-seeded oilseed rape (Brassica napus) and 2 different black-seeded genotypes. A clear correlation was observed between seed colour and ADF content in both crosses. In all 3 populations, a major QTL, with a large effect on both seed colour and ADF in multiple environments, was detected at the same position on chromosome N18. In YE1, a second minor QTL, with a small effect on seed colour but not on ADF content, was localized on chromosome N1. In YE2, no QTL was observed on N1; however, 2 minor seed-colour loci were localized to N15 and N5. A second major QTL for ADF was localized in YE1 on N13; in YE2, no other QTLs for ADF were detected. Combined QTL and segregation data for seed colour and ADF content in the different populations suggest that a partially dominant B. napus gene for seed colour on N18 contributes to a reduction in fibre content in different yellow-seeded B. napus genotypes. The other QTLs that were identified appear to represent different genes in the 2 yellow-seeded rapeseed sources, which, in each case, affect only fibre content or seed colour, respectively. Potential candidate genes and implications for marker-assisted breeding of oilseed rape with reduced seed dietary fibre content are discussed. PMID- 17426766 TI - Seed and agronomic QTL in low linolenic acid, lipoxygenase-free soybean (Glycine max (L.) Merrill) germplasm. AB - Linolenic acid and seed lipoxygenases are associated with off flavours in soybean products. F5 recombinant inbred lines (RILs) from a cross between a low linolenic acid line (RG10) and a seed lipoxygenase-free line (OX948) were genotyped for simple sequence repeats (SSR), random amplified polymorphic DNA (RAPD), sequence tagged sites (STS), and cleaved amplified polymorphic sequence (CAPS) markers and evaluated for seed and agronomic traits at 3 Ontario locations in 2 years. One hundred twenty markers covering 1247.5 cM were mapped to 18 linkage groups (LGs) in the soybean composite genetic map. Seed lipoxygenases L-1 and L-2 mapped as single major genes to the same location on LG G13-F. L-3 mapped to LG G11-E. This is the first report of a map position for L-3. A major quantitative trait locus (QTL) associated with reduced linolenic acid content was identified on LG G3-B2. QTLs for 12 additional seed and agronomic traits were detected. Linolenic acid content, linoleic acid content, yield, seed mass, protein content, and plant height QTL were present in at least 4 of 6 environments. Three to 8 QTLs per trait were detected that accounted for up to 78% of total variation. Linolenic acid and lipoxygenase loci did not overlap yield QTL, suggesting that it should be possible to develop high-yielding lines resistant to oxidative degradation by marker-assisted selection (MAS). PMID- 17426767 TI - Loci affecting flowering time in oat under short-day conditions. AB - Flowering time (or days to heading) is an important characteristic in crop plants that affects adaptation to cropping cycles and growing seasons. The objectives of this study were to identify molecular markers associated with flowering time in 3 oat populations developed from Brazilian oat varieties, and to compare their map locations with those of other loci that might influence flowering time. Flowering time was studied in recombinant inbred lines from 3 hexaploid oat populations: UFRGS 8 x Pc68/5*Starter; UFRGS 881971 x Pc68/5*Starter; and UFRGS 8 x UFRGS 930605. Bulked segregant analysis, using amplified fragment length polymorphism, was followed by selective mapping in each population and in a reference population, 'Kanota' x 'Ogle' (KxO). One quantitative trait locus (QTL) with major effects on flowering time was identified in each cross. Comparative mapping showed that a major QTL, with earliness alleles originating from UFRGS 8 and UFRGS 881971, is in a region with close homology to KxO linkage group 17 and to a locus that reportedly confers day-length insensitivity in oat (Di1). This is the first report to identify the map location of the Di1 locus, and putatively confirm the presence of Di1 alleles in new germplasm. Further comparative mapping and the alignment of mapped oat markers with the sequenced rice genome suggest that this QTL and (or) Di1 is orthologous to the Hd1 locus in rice and the CONSTANS gene in Arabidopsis and other species. A different QTL with major effects segregated in the UFRGS 8 x UFRGS 930605 cross, where the early-flowering allele for Di1 was probably fixed. Two additional QTLs with smaller effects were identified in the UFRGS 8 x Pc68/5*Starter population. These results suggest that the Brazilian oat line UFRGS 8 contains an optimal set of alleles conditioning earliness under the short-day conditions of the Brazilian winter growing season, and that molecular selection could be used to introgress these alleles into other breeding material. PMID- 17426768 TI - Genotype distribution of the prion protein gene (PRNP) promoter polymorphisms in Korean cattle. AB - Recently, an association between bovine spongiform encephalopathy (BSE) and insertion/deletion (indel) polymorphisms in the bovine prion protein gene (PRNP) promoter region has been reported in German cattle. These PRNP polymorphisms cause changes in PRNP expression and are thought to play an important role in BSE susceptibility. BSE has been reported in British and Japanese Holstein cattle but has not been diagnosed in Hanwoo cattle (Bos taurus coreanae) up to now. These results prompted us to investigate the genotype distributions of these PRNP promoter polymorphisms in 107 Hanwoo cattle and 52 Holstein cattle and compare the results with those of previous studies. A significant difference (P=0.0249) in allele frequency of the 23 bp indel polymorphism was observed between Hanwoo and the BSE-affected German cattle previously investigated. There were no significant differences in the genotype (P=0.2095) or allele (P=0.8875) frequencies of the 12 bp indel polymorphism between Hanwoo and BSE-affected German cattle. Interestingly, the genotype and allele frequencies of the 23 bp indel polymorphism in Korean Holsteins were very similar to those previously reported for BSE-affected German cattle and healthy US cattle sires. PMID- 17426769 TI - Molecular characterization and chromosome-specific TRAP-marker development for Langdon durum D-genome disomic substitution lines. AB - The aneuploid stocks of durum wheat (Triticum turgidum L. subsp. durum (Desf.) Husnot) and common wheat (T. aestivum L.) have been developed mainly in 'Langdon' (LDN) and 'Chinese Spring' (CS) cultivars, respectively. The LDN-CS D-genome chromosome disomic substitution (LDN-DS) lines, where a pair of CS D-genome chromosomes substitute for a corresponding homoeologous A- or B-genome chromosome pair of LDN, have been widely used to determine the chromosomal locations of genes in tetraploid wheat. The LDN-DS lines were originally developed by crossing CS nulli-tetrasomics with LDN, followed by 6 backcrosses with LDN. They have subsequently been improved with 5 additional backcrosses with LDN. The objectives of this study were to characterize a set of the 14 most recent LDN-DS lines and to develop chromosome-specific markers, using the newly developed TRAP (target region amplification polymorphism)-marker technique. A total of 307 polymorphic DNA fragments were amplified from LDN and CS, and 302 of them were assigned to individual chromosomes. Most of the markers (95.5%) were present on a single chromosome as chromosome-specific markers, but 4.5% of the markers mapped to 2 or more chromosomes. The number of markers per chromosome varied, from a low of 10 (chromosomes 1A and 6D) to a high of 24 (chromosome 3A). There was an average of 16.6, 16.6, and 15.9 markers per chromosome assigned to the A-, B-, and D-genome chromosomes, respectively, suggesting that TRAP markers were detected at a nearly equal frequency on the 3 genomes. A comparison of the source of the expressed sequence tags (ESTs), used to derive the fixed primers, with the chromosomal location of markers revealed that 15.5% of the TRAP markers were located on the same chromosomes as the ESTs used to generate the fixed primers. A fixed primer designed from an EST mapped on a chromosome or a homoeologous group amplified at least 1 fragment specific to that chromosome or group, suggesting that the fixed primers might generate markers from target regions. TRAP-marker analysis verified the retention of at least 13 pairs of A- or B-genome chromosomes from LDN and 1 pair of D-genome chromosomes from CS in each of the LDN-DS lines. The chromosome specific markers developed in this study provide an identity for each of the chromosomes, and they will facilitate molecular and genetic characterization of the individual chromosomes, including genetic mapping and gene identification. PMID- 17426770 TI - Genetic mapping of agronomic traits in false flax (Camelina sativa subsp. sativa). AB - The crucifer oilseed plant false flax (Camelina sativa subsp. sativa) possesses numerous valuable agronomic attributes that make it attractive as an alternative spring-sown crop for tight crop rotations. The oil of false flax is particularly rich in polyunsaturated C18-fatty acids, making it a valuable renewable feedstock for the oleochemical industry. Because of the minimal interest in the crop throughout the 20th century, breeding efforts have been limited. In this study, a genetic map for C. sativa was constructed, using amplified fragment length polymorphism (AFLP) markers, in a population of recombinant inbred lines that were developed, through single-seed descent, from a cross between 'Lindo' and 'Licalla', 2 phenotypically distinct parental varieties. Three Brassica simple sequence repeat (SSR) markers were also integrated into the map, and 1 of these shows linkage to oil-content loci in both C. sativa and Brassica napus. Fifty five other SSR primer combinations showed monomorphic amplification products, indicating partial genome homoeology with the Brassica species. Using data from field trials with different fertilization treatments (0 and 80 kg N/ha) at multiple locations over 3 years, the map was used to localize quantitative trait loci (QTLs) for seed yield, oil content, 1000-seed mass, and plant height. Some yield QTLs were found only with the N0 treatment, and might represent loci contributing to the competitiveness of false flax in low-nutrient soils. The results represent a starting point for future marker-assisted breeding. PMID- 17426771 TI - Mapping of major spot-type and net-type net-blotch resistance genes in the Ethiopian barley line CI 9819. AB - Net blotch of barley (Hordeum vulgare L.), caused by the fungal phytopathogen Pyrenophora teres Drechs. f. teres Smedeg., constitutes one of the most serious constraints to barley production worldwide. Two forms of the disease, the net form, caused by P. teres f. teres, and the spot form, caused by P. teres f. maculata, are differentiated by the type of symptoms on leaves. Several barley lines with major gene resistance to net blotch have been identified. Earlier, one of these was mapped in the Rolfi x CI 9819 cross to barley chromosome 6H, using a mixture of 4 Finnish isolates of P. teres f. teres. In this study, we used the same barley progeny to map resistance to 4 spot-type isolates and 4 net-type isolates of P. teres. With all net-type isolates, a major resistance gene was located on chromosome 6H, in the same position as described previously, explaining up to 88% of the phenotypic variation in infection response in the progeny. We designate this gene Rpt5. Several minor resistance genes were located on chromosomes 1H, 2H, 3H, 5H, and 7H. These minor genes were not genuinely isolate-specific, but their effect varied among isolates and experiments. When the spot-type isolates were used for infection, a major isolate-specific resistance gene was located on chromosome 5H, close to microsatellite marker HVLEU, explaining up to 84% of the phenotypic variation in infection response in the progeny. We designate this gene Rpt6. No minor gene effects were detected in spot-type isolates. The Ethiopian 2-rowed barley line CI 9819 thus carries at least 2 independent major genes for net-blotch resistance: Rpt5, active against net-type isolates; and Rpt6, active against specific spot-type isolates. PMID- 17426772 TI - A genetic analysis of seed and berry weight in grapevine. AB - Fruit size and seedlessness are highly relevant traits in many fruit crop species, and both are primary targets of breeding programs for table grapes. In this work we performed a quantitative genetic analysis of size and seedlessness in an F1 segregating population derived from the cross between a classical seeded (Vitis vinifera L. 'Dominga') and a newly bred seedless ('Autumn Seedless') cultivar. Fruit size was scored as berry weight (BW), and for seedlessness we considered both seed fresh weight (SFW) and the number of seeds and seed traces (SN) per berry. Quantitative trait loci (QTL) analysis of BW detected 3 QTLs affecting this trait and accounting for up to 67% of the total phenotypic variance. QTL analysis for seedlessness detected 3 QTLs affecting SN (explaining up to 35% of total variance) and 6 affecting SFW (explaining up to 90% of total variance). Among them, a major effect QTL explained almost half of the phenotypic variation for SFW. Comparative analysis of QTLs for these traits reduced the number of grapevine genomic regions involved, one of them being a major effect QTL for seedlessness. Association analyses showed that microsatellite locus VMC7F2, closely linked to this QTL, is a useful marker for selection of seedlessnes. PMID- 17426773 TI - Mapping of FHB resistance QTLs in tetraploid wheat. AB - Triticum turgidum L var. durum is known to be particularly susceptible to infection by Fusarium graminearum, the causal agent for Fusarium head blight (FHB), which results in severe yield losses and grain contaminated with mycotoxins. This research was aimed at identifying FHB resistance in tetraploid wheat and mapping the location of FHB resistance genes. A tetraploid cross of durum wheat ('Strongfield') x Triticum carthlicum ('Blackbird') was used to generate a doubled-haploid (DH) population. This population was evaluated for type II resistance to F. graminearum in replicated greenhouse trials, in which heads were innoculated and the percent of infected spikelets was determined 21 days later. The population was also genotyped with microsatellite markers to construct a map of 424 loci, covering 2 052 cM. The FHB reaction and genotypic data were used to identify FHB resistance quantitative trait loci (QTLs). It was determined that 2 intervals on chromosomes 2BL and 6BS controlled FHB resistance in this tetraploid cross. The FHB resistance allele on chromosome 2BL (r2=0.26, logarithm of odds (LOD)=8.5) was derived from 'Strongfield', and the FHB resistance allele on chromosome 6BS (r2=0.23, LOD=6.6) was derived from 'Blackbird'. Two other loci, on chromosomes 5AS and 2AL, were shown to regulate FHB infection and to have an epistatic effect on the FHB resistance QTL on chromosome 6BS. Further, the FHB resistance QTL peak on chromosome 6BS was clearly coincident with the known FHB resistance gene Fhb2, derived from Sumai 3. The results show that FHB resistance can be expressed in durum wheat, and that T. carthlicum and Triticum aestivum likely share a common FHB resistance gene on chromosome 6BS. PMID- 17426774 TI - Differential responses of Coffea arabica L. leaves and roots to chemically induced systemic acquired resistance. AB - Coffea arabica is susceptible to several pests and diseases, some of which affect the leaves and roots. Systemic acquired resistance (SAR) is the main defence mechanism activated in plants in response to pathogen attack. Here, we report the effects of benzo(1,2,3)thiadiazole-7-carbothioic acid-S-methyl ester (BTH), a SAR chemical inducer, on the expression profile of C. arabica. Two cDNA libraries were constructed from the mRNA isolated from leaves and embryonic roots to create 1587 nonredundant expressed sequence tags (ESTs). We developed a cDNA microarray containing 1506 ESTs from the leaves and embryonic roots, and 48 NBS-LRR (nucleotide-binding site leucine-rich repeat) gene fragments derived from 2 specific genomic libraries. Competitive hybridization between untreated and BTH treated leaves resulted in 55 genes that were significantly overexpressed and 16 genes that were significantly underexpressed. In the roots, 37 and 42 genes were over and underexpressed, respectively. A general shift in metabolism from housekeeping to defence occurred in the leaves and roots after BTH treatment. We observed a systemic increase in pathogenesis-related protein synthesis, in the oxidative burst, and in the cell wall strengthening processes. Moreover, responses in the roots and leaves varied significantly. PMID- 17426775 TI - Microsatellite markers are powerful tools for discriminating among olive cultivars and assigning them to geographically defined populations. AB - Twelve simple sequence repeat (SSR) loci were used to differentiate among 118 cultivars sampled in several countries of the Mediterranean basin and to analyze the genetic structure of olive cultivar gene pools. The markers were found to have high discrimination power. On average, with a single assay it was possible to discriminate 96% of the pairwise comparisons and, with a combination of 3 loci, virtually all cultivars were distinguished. The SSR markers were also tested for their ability to assign cultivars to their geographic population of origin. A selection of 6 loci was found to maximize assignment accuracy, correctly reallocating up to 75.4% of cultivars to their population of origin. Because of the confusion surrounding the origin of most olive cultivars, their molecular identification and ascertainment of origin will be extremely useful for germplasm management and breeding. PMID- 17426776 TI - Genomic distribution of MITEs in barley determined by MITE-AFLP mapping. AB - Miniature inverted-repeat transposable elements (MITEs) represent a large superfamily of transposons that are moderately to highly repetitive and frequently found near or within plant genes. To elucidate the organization of MITEs in the barley genome, MITEs were integrated into the genetic map of barley. In this report, we describe the use of MITEs in amplified fragment length polymorphism (AFLP) mapping, and demonstrate their superiority over conventional AFLP mapping. Barley MITEs include members of the Stowaway, Barfly, and Pangrangja families. By amplifying the flanking sequences of these MITEs, a total of 214 loci were mapped from a population of 93 doubled-haploid segregating individuals between Hordeum vulgare ssp. vulgare and H. vulgare ssp. spontaneum. The 214 MITE-AFLP and 40 anchor simple sequence repeat (SSR) loci were distributed on 7 linkage groups, covering a total map distance of 1 165 cM. The average marker density on each chromosome ranged between 3.4 and 9.6 cM per locus. Only 1 MITE-based locus was frequently found to be associated with MITE loci from the same family, resulting in clusters in chromosomal subregions. In barley, it will be possible to cover the entire genome with a limited set of MITE based primers and to build highly dense maps of specific regions. PMID- 17426777 TI - Cytogenetic analysis of Anaecypris hispanica and its relationship with the paternal ancestor of the diploid-polyploid Squalius alburnoides complex. AB - The karyotype of the endangered fish Anaecypris hispanica was revisited using advanced cytogenetic techniques to elucidate its putative relationship with the paternal ancestor of the hybrid complex Squalius alburnoides and to clarify some of the recently described cytogenetic patterns of the complex. The results of chromomycin A3 and Ag staining, as well as fluorescent in situ hybridization with 28S and 5S rDNA and the (TTAGGG)n telomeric probes, were compared with the patterns observed in specimens of the all-male nonhybrid lineage of S. alburnoides complex, which is considered to reconstitute the nuclear genome of the probably extinct paternal ancestor. Several cytogenetic features observed in A. hispanica specimens were indeed shared by S. alburnoides nuclear nonhybrid males, supporting the hypothesis of a close evolutionary link between A. hispanica and the paternal ancestor of the complex. The genomic rearrangements involving 28S rDNA sites previously described in the S. alburnoides complex and in its maternal ancestor (S. pyrenaicus) were not detected in A. hispanica; they are, therefore, probably due to mechanisms related to hybridization and polyploidy. PMID- 17426778 TI - Complex genome rearrangements reveal evolutionary dynamics of pericentromeric regions in the Triticeae. AB - Most pericentromeric regions of eukaryotic chromosomes are heterochromatic and are the most rapidly evolving regions of complex genomes. The closely related genomes within hexaploid wheat (Triticum aestivum L., 2n=6x=42, AABBDD), as well as in the related Triticeae taxa, share large conserved chromosome segments and provide a good model for the study of the evolution of pericentromeric regions. Here we report on the comparative analysis of pericentric inversions in the Triticeae, including Triticum aestivum, Aegilops speltoides, Ae. longissima, Ae. searsii, Hordeum vulgare, Secale cereale, and Agropyron elongatum. Previously, 4 pericentric inversions were identified in the hexaploid wheat cultivar 'Chinese Spring' ('CS') involving chromosomes 2B, 4A, 4B, and 5A. In the present study, 2 additional pericentric inversions were detected in chromosomes 3B and 6B of 'CS' wheat. Only the 3B inversion pre-existed in chromosome 3S, 3Sl, and 3Ss of Aegilops species of the Sitopsis section, the remaining inversions occurring after wheat polyploidization. The translocation T2BS/6BS previously reported in 'CS' was detected in the hexaploid variety 'Wichita' but not in other species of the Triticeae. It appears that the B genome is more prone to genome rearrangements than are the A and D genomes. Five different pericentric inversions were detected in rye chromosomes 3R and 4R, 4Sl of Ae. longissima, 4H of barley, and 6E of Ag. elongatum. This indicates that pericentric regions in the Triticeae, especially those of group 4 chromosomes, are undergoing rapid and recurrent rearrangements. PMID- 17426779 TI - Polymorphisms in CCL2&CCL5 chemokines/chemokine receptors genes and their association with diseases. AB - BACKGROUND: Chemokines and chemokine receptors are major mediators of leukocyte trafficking into the sites of the immune response. They participate in defence against microbial infection, in Th1/Th2 polarization of the immune response, allograft rejection and angiogenesis/angiostasis as well as in tumorigenesis and metastasis. To date, several functional polymorphisms of chemokine and chemokine receptor genes have been discovered that are able to deregulate chemokine system and, therefore, they may interfere with the pathogenesis of a large number of inflammatory and other diseases. In this review we focus on the known polymorphisms of two chemokines: CCL2, CCL5 and their corresponding receptors (CCR2, CCR5) and we also discuss their associations with susceptibility and progression to selected immune-mediated diseases. METHODS AND RESULTS: Based on relevant literature this article gives a short overview of case-control and family studies regarding effect of the genetic factors on diseases such as coronary artery disease, systemic lupus erythematosus, diabetes mellitus, lung diseases and others. CONCLUSION: Recent advance in the identification of chemokine genetic background of the diseases could provide opportunity for pharmacological treatment. However, we need more information about posttranscriptional events to understand functional relevance of polymorphisms and to discovery new avenues to blocking disease development. PMID- 17426780 TI - MicroRNA biogenesis, functionality and cancer relevance. AB - BACKGROUND: MicroRNAs (miRNA) are small non-coding RNAs that negatively regulate gene expression in a sequence- specific manner. Post-transcriptional silencing of target genes by miRNA occurs either by specific cleavage of homologous mRNA or by specific inhibition of protein synthesis. MiRNAs are essential regulators of various processes such as proliferation, differentiation, development, cell death and interaction between virus and host cell. AIM: The aim of this paper is to summarize the main findings from research on miRNA biogenesis, functionality and cancer relevance. METHOD: A narrative literature review of all of the relevant papers known to the authors was conducted. RESULTS: Several human diseases including cancer are associated with aberrant regulation of miRNAs expression or deficiency in miRNA biogenesis. Analysis of miRNA expression signatures can serve as a valuable tool for cancer classification, diagnostics and prediction of tumor behavior. CONCLUSIONS: There has been demonstrated a possibility to use these microRNA signatures for a specific cancer classification with potential predictive and therapeutic value. The known data provide evidence that microRNAs may open new ways for cancer diagnosis, prognosis estimation and therapy. PMID- 17426781 TI - Present state of evaluating malnutrition in the elderly - analysing indicators. AB - OBJECTIVES: Malnutrition in old age is a significant problem. The study presents a survey of the basic methods used in the diagnosis of malnutrition in the elderly such as nutritional anamnesis, anthropometrical, laboratory a functional examinations and indicating specific differences as compared with a younger population. METHODS: We conducted a search of Czech and foreign literature (using PubMed, Medline) focused on diagnostic methods and examinations of nutritional markers especially by geriatric patients published from 1994 to 2006. RESULTS AND CONCLUSIONS: The examinations we presented are simple, non-invasive methods, which can we use especially for out-patient monitoring of geriatric patients to recognize nutrition disorders. Timely diagnosis of malnutrition in the elderly can prevent patients from worsing self-sufficiency and quality of life of the seniors, increasing the risk of institutionalisation, prolonging hospitalisations due to increased rates of complications and increasing the treatment costs and mortality of geriatric patients. PMID- 17426782 TI - Cortical plasticity and its implications for focal hand dystonia. AB - BACKGROUND: The exact origin of focal dystonias has not been elucidated so far. Aberrant plasticity of the brain cortex is suspected to be a crucial factor in the development of this group of movement disorders. The aim of this article is to summarize recent findings on the etiopathogenesis of focal hand dystonias with a focus on the role of abnormal cortical plasticity. METHODS AND RESULTS: A search of the literature mainly from 1995 to 2005 was done using the PubMed and Ovid search engines. English-language articles were identified using the following keywords: focal hand dystonia or writer's cramp and cortical plasticity, sensorimotor, imaging. Additional references were found through bibliography reviews of relevant articles. The data from neurophysiological and imaging studies, as well as clinical observation, in focal hand dystonia suggest multiple failures at different levels of the somatosensory and motor systems, particularly in the brain cortex. This disorders lead to attenuation of inhibitory and fortification of excitatory processes. CONCLUSIONS: The emerging theory presumes that a maladaptive plasticity of brain cortex with abnormal sensorimotor intergration can evolve in predisposed individuals. Consequent methods of management of focal hand dystonias are outlined. PMID- 17426783 TI - Excipients and their role in absorption: Influencing bioavailability of cyclosporine by triglycerides and polyglycerol esters. AB - AIMS: The goal of the experiment was to verify bioavailability of immunosuppressive drug cyclosporine after oral administration, to determine, in particular, effect of triglycerides and polyglycerol esters on bioavailability of this lipophilic, poorly soluble drug. METHODS: 1) Absorption of cyclosporine A from soya oil and polyglycerol-3-oleate was tested after intra-duodenal application to rats. This method enable to administer dispersion directly to the site of absorption and avoid problems with potential precipitation of active substances in the stomach. Samples were pre-dispersed in water and diluted on concentration 1mg/ml prior administration. In defined time intervals a blood sample was taken in a volume of 0.5 ml from the cannuled carotid. Blood was sampled in time 5, 15, 30, 45, 60, 90, 120, 180 and 240 minutes after application. For the comparison, Equoral oral solution diluted and pre-dispersed in the same manner was used. 2) 100 mg of cyclosporin in form of 1% dispersion in water was administered orally to dogs under fasting condition. The blood level of cyclosporine was evaluated from samples taken in time 0.5, 1, 1.5, 2, 2.5, 3, 5, 8, 12 and 24 hours following application. Formulations containing different ratio of polyglycerol esters / olive oil were compared. RESULTS: The experiments conducted indicate that cyclosporine is 19x more available from polyglycerol-3 oleate than from soya oil. When applying cyclosporine in polyglycerol-3-oleate the average maximum blood level is 10x higher then in application of cyclosporine in oil. If polyglycerols are fully substituted with plant oils in the formula observed, its pharmacokinetic parameters decrease to 1/10 of the initial values. CONCLUSIONS: The right selection of a type of excipient accompanying cyclosporine affects significantly cyclosporine availability and thus its efficiency. PMID- 17426784 TI - Evaluation of urine N1,N12-Diacetylspermine as potential tumor marker for urinary bladder cancer. AB - BACKGROUND: N1,N12-diacetylspermine, a diacetylpolyamine which was recently identified in urine, appeared to be a useful tumor marker for a number of cancers. No valid data on urine diacetylspermine concentration in patients with urinary bladder cancer exist. AIM: Evaluation of urine N1,N12-diacetylspermine concentrations in individuals with urinary bladder cancer. METHODS: Urine samples were used from 36 patients with urothelial tumors of the urinary bladder and from 30 patients with benign urological diseases. Urine was collected before cystoscopy. Enzyme-linked immunoabsorbent assays (ELISA) were performed for diacetylspermine from urine. RESULTS: Urine diacetylspermine did not differentiate in individuals with urinary bladder cancer from controls (medians 171.5 vs 143.8, p = 0.64). Its efficacy for urinary bladder cancer detection was not shown. CONCLUSIONS: Urine N1,N12-diacetylspermine is probably not a useful marker for urinary bladder cancer. PMID- 17426785 TI - Our experiences with measurement of new potential biomarkers in the diagnosis of latent forms of myocardial ischemia. AB - BACKGROUND: The need for a laboratory marker of myocardial ischemia has been alluded to for at least the last decade. AIM: The aim of this study was to evaluate the diagnostic importance of the myosin light chain-1 (MLC-1), clusterin and Reg-Ialpha in patients with suspected myocardial ischemia. METHODS: A group of 176 at high-risk for myocardial ischemia subjects was evaluated and divided into two subgroups using myocardial SPECT (Single Photon Emission Computed Tomography) - individuals with and without signs of myocardial ischemia. Laboratory markers in venous blood were repeatedly examined in all subjects: a) immediately prior to SPECT: C-reactive protein, Haemoglobin, Hematocrite, Lactate, MLC-1, Clusterin, Reg-Ialpha b) at subjective maximum: Hb, Htc, lactate, MLC-1, Clusterin, Reg-Ialpha c) 30 min after stress levels reached their peak: MLC-1, Clusterin, Reg-Ialpha and d) 60 min after peak stress levels: MLC-1, Clusterin, Reg-Ialpha. RESULTS: Patients were divided into subgroups according to their positive and negative SPECT results (positive: n = 37; negative: n = 139). MLC-1 values were different for all 4 blood collections. An increase in MLC-1 > 2.2 mg/l showed 64 % sensitivity and 88 % specificity for the diagnosed presence of myocardial ischemia (AUC 0.81; LR+ 5.9; PPV+ 68 % and NPV- 87 %). There was no significant difference between the groups in terms of Clusterin and Reg-Ialpha for any of the sampling periods. CONCLUSIONS: High diagnostic efficacy of detectable MLC-1 was shown for the diagnosis of latent myocardial ischemia. Measurement of serum Clusterin or Reg-Ialpha did not sufficient for the diagnosis of latent myocardial ischemia. PMID- 17426786 TI - Detection of Aspergillus spp. in biological samples by real-time PCR. AB - BACKGROUND: Recently, the proportion of invasive infections caused by the filamentous fungi of the Aspergillus genus are growing in immunocompromised persons particularly in transplant recipients and neutropenic patients. Unfortunately, laboratory diagnostics of invasive aspergillosis remains extremely difficult, mainly with regard to the sensitivity of the methods and to the correct interpretation of the results in particular. AIM: The aim of this work was to design a standard and reproducible Aspergillus DNA detection method and its validation. The second aim was to practically use this method for diagnosis of Aspergillus DNA in various samples in patients. METHOD: Real-time PCR with two hybridization probes. Amplification and on-line quantification was carried out on a LightCycler 1.5 Instrument. RESULTS: Specificity of the reaction was tested for A. fumigatus, A. flavus, A. niger and A. terreus, and its sensitivity was determined at 5 copies per ml. The reproducibility of the results was comparable to other methods, reported in the literature. Applicability of the real-time PCR was assessed for detection of Aspergillus DNA in 354 various clinical samples taken from 179 patients at risk of invasive aspergillosis over the period of 33 months. Of 354 samples 103 (29.10 %) taken from 65 patients (36.31 %) were evaluated as positive. Over one year, the percentage of positive samples was mostly about 30 % or less per month. CONCLUSIONS: Our results demonstrate the high sensitivity, specificity and reproducibility of this technique, and its usefulness for rapid laboratory diagnosis of invasive aspergillosis. PMID- 17426787 TI - Quantitative and qualitative analysis of proteins in rat peripheral nerves predegenerated for 7 days. AB - OBJECTIVES: In contrast to peripheral nerves, central neurons do not regrow spontaneously after injury. Our previous studies showed that transplantation of degenerating peripheral nerves or their extracts can induce regeneration in the injured central nervous system. Non-predegenerated nerves show much weaker neurotrophic activity. The aim of the present work was to examine quantitatively and qualitatively the protein composition of rat sciatic nerve extracts. MATERIAL AND METHODS: The experiments were carried out on male Wistar C rats. Distal fragments were collected immediately after transection or after 7 day-long predegeneration. The nerves were homogenized, centrifuged and ultracetrifuged. Extracts were analyzed by means of two-dimensional electrophoresis. RESULTS: The two-dimensional electrophoresis showed 69 protein subfractions with isoelectric points ranging from 4.2 to 7.0 pH and molecular weight ranging from 13.5 kDa to 335.4 kDa in extracts obtained from nonpredegenerated nerves. In predegenerated nerve extracts 114 subfractions with isoelectric points ranging from 4.2 to 7.4 pH and molecular weight from 21.1 kDa to 335.4 kDa were found. Fractions: 25.5 kDa, 31.6 kDa, 36 kDa, 38.4 kDa, 42.4 kDa, 46.6 kDa, and 50.5 kDa showed significant increase and two fractions: 68.5 kDa and 335.4 kDa demonstrated significant decrease in the number of subfractions in predegenerated nerves. Fractions 160.8 kDa, 236.1 kDa, and 5 fractions below 21.1 kDa were present only in extracts from non-predegenerated nerves. CONCLUSIONS: In conclusion, the results of our study demonstrate that the most intense changes in protein composition in degenerating nerves take place in low molecular weight fractions. PMID- 17426788 TI - Prognostic value of hMLH1 and hMSH2 immunohistochemical expression in non-small cell lung cancer. A tissue microarray study. AB - BACKGROUND: hMLH1 and hMSH2 genes are both known to play a role in DNA mismatch repair. Nonetheless, the clinical significance of hMLH1 and hMSH2 protein expression in lung cancers remains unclear. AIM: The aim of this study was to investigate the immunohistochemical expression of hMLH1 and hMSH2 proteins in tumor specimens from 179 non-small cell lung cancer (NSCLC) patients using a tissue microarray technique and to correlate these results with other clinicopathological variables, including the disease specific and overall survivals. METHOD: hMLH1 and hMSH2 protein expression was evaluated by immunohistochemistry using monoclonal antibodies G168-728 for hMLH1 and FE11 for hMSH2 protein expression analysis. The Pearson chi2 test was used to compare the hMLH1 and hMSH2 alterations among the cases and between various clinical and laboratory variables. P < or = 0.05 was considered statistically significant. RESULTS: Alteration of hMLH1 and hMSH2 protein expression was observed in 10 % of patients. No significant correlation was found between the protein expression and patient age, smoking status, tumor histology or disease stage and disease free and overall survival. CONCLUSIONS: Alterations in the expression of hMLH1 and hMSH2 proteins did not have any prognostic value in stage III. NSCLC patients. PMID- 17426789 TI - The relationship between morphological indicators of human body and posture. AB - BACKGROUND: Posture and Stability Control is a process of maintaining balance and position of the body and its parts in a constantly changing environment. It is an important regulatory mechanism of the body, because it precedes movement, is upon the completion of which this system tries to maintain the body position. AIM: The purpose of this long term study is to determinate by special testing which morphological indicators and how influence postural stability and its control. METHOD: This study used testing with NeuroCom Balance-12 protocols, anthropometric measuring - 32 values, anamnestic questionnaire. For this article we analyzed one chosen protocol: Weight Bearing Squat Test. RESULTS: In this research, 41 individuals took part in it. 22 (53.66 %) were female, 19 (46.34 %) were male. Interesting statistically significant coherences were discovered during analysis of Weight Bearing Squat protocol on level p < 0.01, p < 0.05 and the Student's two-optional T-test proved statistically significantly higher values of Left Body Weight erect position and Left Body Weight 30 degrees knee flexion in men in comparison with women. CONCLUSIONS: This study shows us a direction and we may consider it a convenient base for more specific continuation in this problematic area. PMID- 17426790 TI - Early diagnosis of DIC development into the overt phase. AB - BACKGROUND: Disseminated intravascular coagulation (DIC) is a syndrome characterized by over-activation of intravascular coagulation, accompanied by consumption of coagulation factors. It is a pathologic systemic inflammatory response accompanied by release of coagulatory substances into the circulation. AIM: The aim of this paper is to emphasize the importance of early the early diagnosis of DIC and to present a case report of a successful early diagnosis and treatment. METHOD: A case report giving a detailed account of clinical and laboratory presentation of a case of DIC in a patient with sepsis. Relevant laboratory results and other tests are presented, treatment is described in detail. CONCLUSIONS: DIC remains an important and potentially life-threatening complication of severe disease and states. Early diagnosis is vital for the success of treatment, and in severely ill patients, the possibility of DIC should always be kept in mind in order to detect early signs and initiate treatment as soon as possible. PMID- 17426791 TI - Selected pregnancy variables in women with placental abruption. AB - OBJECTIVE: The aim of this study was to investigate risk factors for placental abruption and to determine if anamnestic variables such as inherited thrombosis or recurrent fetal loss might be used as a predictor for placental abruption. METHODS: A retrospective case-control study at the University Hospital, Palacky University, Olomouc, Czech Republic. One hundred and eighty women with placental abruptio out of 20,175 deliveries (0.79 %) who were compared to 196 unselected pregnant women. A detailed anamnesis was taken. RESULTS: Compared to controls, women with placental abruptio had a 12-fold increased prevalence of prior recurrent fetal loss and a 6-fold increased prevalence of inherited thrombosis. CONCLUSIONS: We found that recurrent fetal loss, and inherited thrombosis may be significant risk factors for placental abruptio. PMID- 17426792 TI - Screening for chromosomal anomalies in the first trimester: a report on the first year of prospective screening for chromosomal anomalies in the first trimester in the Czech Republic. AB - BACKGROUND: The increase in maternal age in recent years has intensified the effort to develop early non-invasive methods for screening for trisomy 21 and other chromosomal abnormalities in prenatal diagnosis. In the first trimester of pregnancy, maternal age, fetal nuchal translucency (NT), maternal levels of free beta- human chorionic gonadotropin (beta-hCG) and pregnancy-associated plasma protein-A (PAPP-A) are used as screening markers. We evaluated the introduction of this method of screening for the first time in the Czech Republic. METHODS: it is a prospective study for one-year from the beginning of 2004. The risk of trisomy 21(Down's syndrome) was estimated for 686 singleton pregnancies. The specific risk was calculated using the Fetal Medicine Foundation software (FMF) by accredited sonographers. Karyotyping was offered to women with risk >or= 1 in 250. RESULTS: In the population screened 18 % of women were aged 35 and more. We found 2 cases of trisomy 21 and 1 case of trisomy 18 (Edward syndrome) resulting in a detection rate of 100 % for trisomy 21 for a 5 % false positive rate (33 of 683). The maternal age of the detected cases was 30, 38 and 42 years. CONCLUSION: Introduction of the first trimester screening to our clinic, reduced the number of invasive genetic testing from 18 % to 5 %. First trimester screening for trisomy 21 and other aneuploidies has a high sensitivity with a low false positive rate and can be delivered in an efficient manner in a university hospital. PMID- 17426793 TI - Uterine rupture during pregnancy and delivery among women attending the Al Tthawra Hospital in Sana'a City Yemen Republic. AB - BACKGROUND: About 20 percent of the population in developing countries is composed of women of reproductive age. These women face one of the catastrophic risks of pregnancy "uterine rupture". Studies conducted in the developing world give strong evidence that uterine rupture is a major health problem in these countries with the rate being high in rural areas. AIM: The purpose of the study was to estimate the incidence and determine the risk factors and outcome of uterine rupture among women using the referral hospital Al-thawra in Sana'a City, Yemen republic and to extrapolate the data to the whole of Yemen. METHODS: The data was collected retrospectively; by interviewing, examining and following up all the cases of uterine rupture coming to the hospital during a period of 9 months between September 1996 and May 1997. A descriptive analysis and distribution frequency of the commonest causes of uterine rupture in 37 cases are presented taking into account medical, reproductive, health services provided and sociodemographic factors. RESULTS: Incidence of uterine rupture in Yemen was found to be (0.63), obstructed labor 83 %, contracted pelvis 19 %, previous surgery in 48 %, Oxytocine infusion in 42 %. Grand-multiparty was in 65 % and maternal age over 35 years in 50 %. Antenatal care was only in 34 %. CONCLUSION: The high percentage of malpresentation, cephalopelvic disproportion, previous uterine surgery accompanied by the high percentage of use of Oxytocin in this study highlights very clearly the role of this medication in increasing the risk of uterine rupture in Yemen. PMID- 17426794 TI - The relevance of brain natriuretic peptides investigation in various cardiovascular diseases. AB - BACKGROUND: Brain natriuretic peptides are relevant markers of heart impairment. AIM: We investigated the relevance of investiging brain natriuretic peptides (NT proBNP, BNP) in monitoring different types of cardiovascular disease (chronic heart failure due to coronary artery disease, cardiomyopathy, acquired valve disease, congenital heart diseases). METHODS: The NT-proBNP assay (Roche) was performed on 280 patients (mean age 49 years; range 20-89 years) and 48 healthy controls (mean age 43 years; range 13-65 years) and BNP assay (Bayer Shionoria) was performed in a subgroup of 42 patients (mean age 50 years; range 20-79 years). Patients were divided into four groups characterized by severity of heart failure according to the New York Heart Association classification. RESULTS: NT proBNP concentrations differed in patients with cardiovascular diseases from controls (median 371 ng/l versus 41.5 ng/l, p < 0.0001). The cut off value of NT proBNP determined in 280 patients with cardiovascular diseases was at 130 ng/l (AUC-area under curve = 0.93; sensitivity 98 %; specificity 79 %). Comparison of NT-proBNP and BNP values in patients showed significant correlation (r = 0.93; p < 0.0001). NT-proBNP showed significant differences between groups. CONCLUSIONS: Measurement of brain natriuretic peptides is useful and relevant in various types of heart diseases including congenital. PMID- 17426795 TI - Prevalence of inherited thrombophilia in patients with severe ovarian hyperstimulation syndrome. AB - BACKGROUND AND OBJECTIVE: To determine the prevalence of markers of inherited thrombophilia in patients with severe form of ovarian hyperstimulation syndrome (OHSS) DESIGN AND METHODS: Blood samples were analysed for markers of thrombophilia (mutation of factor V - Leiden, mutation of methylentetrahydrofolat reductase (MTHFR) C677T and mutation of prothrombin G20210A). The study group consisted of women who had undergone controlled ovarian hyperstimulation for IVF complicated by severe OHSS (group A, n = 50). Results were compared with two controls groups - women who underwent ovarian hyperstimulation for IVF without developing OHSS (group B, n = 93) and healthy pregnant women with no history of infertility (group C, n = 196). RESULTS: We have found 7 out of 50 patients from group A positive for Leiden mutation (heterozygous) 11 out of 93 from group B (heterozygous), p = 0.71 OR 1.21 (0.39 3.70) and 10 out of 196 from group C (heterozygous), p = 0.03 OR 3.03 (0.97 9.28). Polymorphism of MTHFR 677T gene was detected in 17 out of 50 patients (heterozygous) from group A, in 36 out of 93 patients from group B (35 heterozygous, 1 homozygous) and in 93 out of 196 patients from the group C (86 heterozygous, 7 homozygous) with no statistical significance. CONCLUSIONS: We found an increased prevalence of factor V Leiden mutation in Czech infertile women. However carriers of Leiden mutation had no enhanced risk of development severe form of OHSS during stimulation. PMID- 17426796 TI - Acute severe mitral regurgitation with cardiogenic shock caused by two-step complete anterior papillary muscle rupture during acute myocardial infarction. AB - BACKGROUND: Rupture of a papillary muscle is an infrequent but often fatal mechanical complication of acute myocardial infarction. AIM: The aim of this paper is to present a case report of a 65-year old women with acute severe mitral regurgitation with cardiogenic shock caused by two-step complete anterior papillary muscle rupture during acute myocardial infarction. The transthoracic echocardiography was obtained at the bedside and showed a posterior mitral valve prolapse with a severe mitral regurgitation. During this examination the patient developed acute pulmonary oedema and a consequent cardiogenic shock. Transthoracic echocardiography was then reevaluated and completed. New findings of bi-leaflet mitral flail and progression of massive mitral regurgitation were documented. The complete rupture of a papilary muscle was then considered as a cause of an acute clinical deterioration. Intraoperative findings showed a complete transection of both heads of anterolateral and necrotic regions of basis of posteromedial papillary muscle. CONCLUSIONS: This case confirms the importance of immediate echocardiography in confirming a diagnosis of acute mechanical complications of acute coronary syndromes and this examination is important for the management of a hemodynamically unstable patient. Echocardiography should be done immediately on any patient in whom the diagnosis of mechanical complication of acute coronary syndromes is suspected. PMID- 17426797 TI - An uncommon case of spontaneous resolution of mural thrombus causing symptomatic severe stenosis of the extracranial part of the internal carotid artery. AB - OBJECTIVES: Etiology of ischemic stroke(IS) remains sometimes unexplained in younger subjects. Its well-known causes are atherosclerosis(AS), embolization, arterial dissection, congenital defects, vasculitides, smoking, use of oral contraceptives, coagulopathy and arterial dissection, or vasospasm. CASE REPORT: A 54-year old male subject was examined because of motor disturbance in his left upper extremity. Computerized tomography(CT) of the brain did not detect any focal pathological changes. Utrasonographic(USG) examination revealed severe stenosis of the right internal carotid artery(ICA) caused by hypoechogenic material, the blood velocity beyond the stenosis was 240 cm/s. The USG finding was confirmed by digital subtraction angiography (DSA). Based on the results of these examinations, an acute carotid endarterectomy (CEA) was indicated. During CEA the vascular surgeon observed patency of the ICA with only small calcified AS plaque on its posterior side. Ultrasonographic examination, performed on the 5(th) day, showed a small calcified AS plaque on the posterior side of the right ICA. CONCLUSION: In conclusion, if we exclude the possibility of a false positivity of 2 imaging methods at once, we have encountered a very unusual case of SR of stenosing material in the extracranial part of the ICA, initially causing its severe stenosis manifesting by acute neurological symptomatology. The mechanism of this SR remains unclear and the problem should be further studied, preferably using less invasive AG methods. PMID- 17426798 TI - Bifocal pacing - A novel cardiac resynchronization therapy? Results of bifocal pacing study and review of the current literature. AB - BACKGROUND: Bifocal pacing (BFP) has been proposed as a more feasible alternative of cardiac resynchronization therapy (CRT). AIM: To evaluate BFP in patients with severe congestive heart failure and significant intraventricular conduction delay and to compare it with biventricular pacing (BVP). METHODS: Both echocardiographic examination including tissue Doppler imaging and invasive measurements of cardiac hemodynamics was performed under basal conditions and during BFP and BVP. RESULTS: 50 patients were included: 29 patients with ischemic heart disease (IHD), 21 patients with idiopathic dilated cardiomyopathy (IDCM). Left ventricular (LV) pressure gradient (dp/dt max) increased during BFP compared to the baseline (13.4 %, 95 % CI 9.2-17.6 %, p < 0.0001) and a further increase was achieved during BVP (29.5 %, 95 % CI 23.7-35.4 %, p < 0.0001). A significant correlation was found between the distance of the right ventricular apical and outflow tract leads and percentage of dp/dt max increase in IDCM patients (r = 0.72, p < 0.001), but not in IHD patients. Interventricular mechanical delay (IVMD) decreased in BFP compared to baseline (43 +/- 22 ms vs. 53 +/- 31 ms, p = 0.006). BVP produced even shorter IVMD (22 +/- 19 ms, p < 0.0001). In all patients, the regional systolic contraction times were significantly shortened, corresponding with prolongation of the respective regional diastolic filling times during both BFP (p < 0.05 for all segments) and BVP (p < 0.001 for all segments). The effect of BVP on regional systole shortening was more pronounced. CONCLUSIONS: BFP improves LV hemodynamics by decreasing the inter- and intraventricular conduction delays. The leads in the right ventricle should be placed at the longest achievable distance. BVP is superior to BFP. PMID- 17426799 TI - Possible restorations of the upper extremity motion in tetraplegic patients - 5 year clinical experience. AB - AIM: The aim of the work is to objectify the functional effectiveness of these operations and their influence on the quality of life of handicapped patients. METHOD: The authors evaluate the results of reconstructive surgery restoring hand grip in a group of 15 tetraplegic patients (3 women and 12 men) with complete spinal cord lesion of C5-C7 segments. The average age of patients in the group is 33 (22-50) years old. The reconstructions were performed using tendon transfer and tenodesis in the forearm and hand area. The effectiveness of the transfer was assessed objectively with regard to muscle strength by measuring the restored "thumb-index finger" grip and "into fist" grip. The range of motion achieved was also evaluated. Transfer effectiveness was evaluated on the basis of subjective patients' evaluation. An ADL (activities of daily living) questionnaire by Mohammed's (1992) took into account the effect of the surgery in a whole range of common daily activities. RESULTS: An extended range of daily activities was evident mainly in the fields of: communication, eating and drinking and operations associated with increase in general selfcare of the patient. There was no deterioration of condition in any of the activities. CONCLUSIONS: Up to 80 % of tetraplegic patients are suitable candidates for transfers and, to a certain extent, it is possible to improve the upper limb function. In a partial function restoration of the upper limbs there is immense potential for improvement in the quality of life of these patients. PMID- 17426800 TI - Wear measurement of retrieved polyethylene ABG 1 cups by universal-type measuring microscope and X-ray methods. AB - BACKGROUND: Polyethylene wear is considered a most important part of periprosthetic osteolysis development. Thus, its measurement is central to contemporary orthopaedics. AIMS: The aim of this paper was to compare the accuracy of three radiographic techniques for wear measurement. Secondly, the influence of the abduction angle of the cup on measurement accuracy was investigated. METHODS: Wear was measured manually in 80 patients by a single observer according to the Livermore, Charnley, and Dorr description. A multi component statistical analysis was used to test the hypothesis that the Livermore technique was superior. In vitro data obtained from a Universal-type measuring microscope served as a gold standard. RESULTS: In vitro measurements showed an average linear wear of 0.363 mm per year (0.000-0.939, SD 0.241) with a corresponding volumetric wear rate of 161 mm3 per year (0-467, SD 118.2). The Livermore technique showed the least deviation from the optical reference standard and a superior position from the viewpoint of error analysis but the correlation coefficient was slightly less (r = 0.761) than for the Dorr and Charnley techniques (r = 0.795 and r = 0.778, respectively). In addition, the mean error of the Dorr method differed significantly from zero (p = 0.036). Overall, the Livermore technique was the most accurate method for polyethylene wear measurement regardless of the abduction angle of the cup. CONCLUSIONS: The Livermore technique performed manually was more accurate than the Charnley and Dorr methods. Nevertheless, we consider the Dorr technique an adequate tool for day-to-day wear measurements, mainly due to its simplicity. PMID- 17426801 TI - Use of the Pavlik nail for femoral shortening. AB - BACKGROUND: Selecting the correct type of osteosynthesis has a crucial impact on the final outcome of shortening operations for leg length discrepancy (LLD). One approach, introduced at the beginning of the 1950's, was fixation of femoral shortening osteotomy using the Pavlik intramedullary nail. AIM: The aim of this paper is to draw attention to the outstanding contribution of this great representative of Czech orthopaedics, to compare his method with other approaches to osteosynthesis, and, in the original patient set, to evaluate the long-term results of femoral shortening based on available medical records. MATERIAL AND METHODS: Of 34 operated patients, a subset of 11 underwent femoral shortening and fixation by the Pavlik nail and were evaluated according to the Merle d'Aubigne and Postel score. Preoperative, postoperative and current radiographs are described. RESULTS: The results were: excellent (6), good (3), and fair (2). There was no infection, thrombembolism, or osteosynthesis failure. Heterotopic ossification was recorded. The general validity of Pavlik's technical orthopaedic thinking is confirmed by stable osteosynthesis, and prevalence of excellent and good results. CONCLUSION: This study has confirmed the general validity of Pavlik's technical and orthopaedic thinking. His intramedullary nail was a suitable instrument for achieving stable osteosynthesis after a shortening femoral osteotomy. There was neither nail migration nor bone fragment dislocation. No additional plaster immobilization was used and this promoted early, progressive and rapid rehabilitation oriented towards active exercises. The low complication rate was evidence of correct methodology. Despite the undoubted value of the Pavlik femoral intramedullary nail, this method has never become widespread. PMID- 17426802 TI - Cisatracurium vs. Rocuronium: A prospective, comparative, randomized study in adult patients under total intravenous anaesthesia. AB - AIMS: To compare the pharmacodynamics of cisatracurium and rocuronium-induced neuromuscular block following single dose, allowing either spontaneous or neostigmine-accelerated complete recovery. METHODS: Following the ethics committee approval and informed consent, 120 patients scheduled for elective surgery under TIVA with tracheal intubation were randomized into 4 groups with different cisatracurium (CIS, 0.10 or 0.15 mg.kg(-1)) or rocuronium (ROC, 0.60 or 0.90 mg.kg(-1)) doses administered. For each patient, the onset time for 95 % depression of T1, clinical duration until 25 % recovery, recovery index (T1 from 25 to 75 %) and time from T1 25 % to TOF-ratio 0.9 were determined allowing either spontaneous or induced recovery. RESULTS: The onset times were 277 (SD 58), 220 (46), 91 (16) and 77 (16) s for the CIS 0.10, CIS 0.15, ROC 0.60 and ROC 0.90 groups (p < 0.05), respectively, with lower variability in both ROC groups (p < 0.05). The clinical durations were 42 (7), 52 (7), 35 (11) and 52 (12) min, respectively (p < 0.05 for lower doses). Recovery index was identical in all groups allowing either spontaneous recovery - 15.9 (1.8), 15.5 (1.7), 16.1 (3.7) and 16.1 (4.0) min, or following neostigmine administration - 4.4 (0.9), 4.5 (0.8), 4.3 (0.8) and 4.7 (0.7) min for respective groups. During spontaneous recovery, the variability of DUR25-TOF90 was twice as great for ROC than CIS groups (p < 0.05), while after neostigmine administration it was uniform in all groups. CONCLUSIONS: For equipotent doses, the onset times for CIS were approximately three times longer than for ROC. The average clinical duration for both relaxants ranged from 35 to 52 min with acceptable variability. Neostigmine administration accelerated the recovery and reduced its variability. When allowing for spontaneous recovery, less scatter was demonstrated for both CIS groups than for ROC ones. PMID- 17426803 TI - Pulse wave variability within two short-term measurements. AB - BACKGROUND: Pulse wave analysis permits non-invasive assessment of indices of arterial elasticity. Decreased arterial elasticity results in higher pulse wave velocity and therefore earlier wave reflection. AIM: The aim of this study was to evaluate the short-term variability for measurement of surface arteries. METHOD: For detection of artery wall movement, a plethysmography method was used. The changes in artery volume, affecting the fluid behind the scanning membrane, transfer themselves into pressure changes and move towards the positive input of a sensitive differential pressure transducer. The output is carried to a notebook computer and analysed within the frequency and time domain. To correlate arterial pulse waves, the following parameters were used: relative crest time RCT, interwave distance IWD, dicrotic wave amplitude DWA and dicrotic wave time DWT. These parameters were analysed using Bland-Altman plots. RESULTS: Two data points (5.6 %) fall outside the 2*SD range for both IWD and DWA parameters on posterior tibial artery and one data point (2.8 %) for DWT. Three data points (8.3 %) fall outside the 2*SD range for IWD and two data points for both DWA and DWT on the radial artery. Two data points fall outside the RCT measured on the ulnar artery, and all but one for IWD, DWA and DWT lie within the 2*SD range. CONCLUSIONS: Pulse wave analysis showed reproducibility in a short-term study of all three measured distal arteries and is therefore suitable for use in patients for observing the progression of artery-wall-involving diseases. PMID- 17426804 TI - Possibilities of experimental verification of the validity of urethral pressure profilometry. AB - OBJECTIVES: The aim is to define conditions for exact measurement of urethral pressure profile, to prepare an aparatus and fabricate an artificial urethra for testing measuring catethers. To examine it's qualities by experiment. METHODS: We designed a measuring appliance and a specialized software for measuring and evaluating. An experimental phantom of urethra, made from a part of bull's urethra, was used for testing of measurement's validity with use of different measuring catethers. Measurements were 10x repeated within the same position of the catether. Reproducibility of the measurement and stability of the experimental model was tested. RESULTS: Measuring urethra and experimental phantom of urethra succeeded, it was possible to make well reproducible measurements with different measuring catethers and to evaluate their metrologic abilities and limits. Results of our measurements confirmed appointed theoretical conditions of the measurement. CONCLUSION: Measurements proved that it is possible to simulate the conditions that are similar to physiological conditions and that it is possible to test metrological qualities of compressive profilometry of urethra. PMID- 17426805 TI - The position of a doctor in cases of violence attack commited by a patient from the point of view of juristic theory. AB - There is no doubt, that the medical staff has often to face aggressive patients, their threats and event their violent physical attacks today. More frequently than ever they are faced with the necessity to think over and consider very carefully the way to defend against imminent or persistent attack on them. First, I would attempt to qualify patients' aggressive behaviour - from lesser acts, when the violent attack is not imminent, through threatening with killing, inflicting aggravated bodily harm or causing exceptionally serious damage or enforcement by violence, the threat of violence or the threat of causing another serious damage, to do something, to desist from doing something or tolerate something. to the imminent or persistent attack. Then I would try to describe the best ones of possible ways of self-defence against various levels of aggressive behaviour. PMID- 17426806 TI - Web-based instruction and its impact on the learning activity of medical students: a review. AB - BACKGROUND: The aim of this review was to summarise the experience on implementation of information technology to support the teaching and learning process in medicine. Particular attention was paid to web-based tutorials, their impact on increasing the effectiveness of medical instruction and motivation of students towards self-directed learning. Most of the studies selected for the purpose of the review comprised evaluation of the web-tutorials in view of practical implementation, strengths, weaknesses, and main preferences in comparison with traditional lecture-based education. METHOD AND RESULTS: We searched MEDLINE via PubMed using MeSH term "computer-assisted learning" between 1996 and 2005 and selected for inclusion in this review were studies on the implementation and evaluation of web based tutorials in medical education. Additional related papers were obtained through cross-referencing. We found that overall, students prefer Web tutorials to traditional lecture-based classes for accessibility, ease of use, freedom of navigation, high medical image quality and advantage of repeated practice, that web-based learning has been continually developing and that it is a very important tool in Evidence Based Medicine. CONCLUSIONS: Web based education is an important tool in medical training. It will require transformation in the way medicine is taught from instructor based to self directed learning. It is above all seen as a device for information retrieval and storage. PMID- 17426807 TI - Lipid metabolism in active Crohn's disease: pre-results. AB - BACKGROUND: Crohn's disease (CD) is a chronic relapsing disease. Especially acute period may be associated with metabolic disturbances. Alteration of lipid metabolism has been described in critically ill patients and hypocholesterolemia is associated with poor prognosis. The human organism acquires cholesterol by two principal processes - synthesis de novo, and absorption from the diet. It is possible to assess, using cholesterol synthesis markers (lathosterol) and cholesterol absorption markers (sitosterol, campesterol) the leading form of cholesterol acquisition. AIM: The aim of this study is assess the association between the lipid profile in plasma and the plasma concentration of sterols in active CD patients and in control subjects. METHOD: Routine laboratory tests, CDAI, lipid and non-cholesterol sterols plasma levels were performed on days 3, 14 and 28. The metabolic parameters have been compared with a control cohort of 100 healthy blood donors. RESULTS: Presently, complete data for 8 patients are available The serum total cholesterol, LDL and HDL cholesterol, and triglyceride concentrations were lower in patients with acute Crohn's disease than in the control group. Moreover lathosterol, campesterol and sitosterol concentrations were lower, whereas squalene concentration was higher than in controls. As mentioned above, complete data are not currently available. Therefore statistical analysis has not been finished. CONCLUSION: Our pre-results show substantial abnormalities in the concentrations of plasma lipids and non-cholesterol sterols, which are presented as markers of cholesterol requirement, in patients with acute CD. PMID- 17426808 TI - Microcirculation in the aggregated lymphoid nodules in pig. AB - AIMS: To compare the blood supply of the Peyer's patches in pigs with the already defined rodent one. MATERIALS AND METHODS: Pig tissue was used. Injections of india ink, microscopic examination of the histological preparations stained by a haematoxylin - eosin and scanning electron microscopy of the corrosive casts were used for the depiction of the vessels. RESULTS: A model of the blood microcirculation of PP and its relation to the blood circulation of the small intestine was created. Only the capillaries in the follicles, but not the ascending arterioles as in rats were found. PMID- 17426809 TI - Recalibrated tree of leaf beetles (Chrysomelidae) indicates independent diversification of angiosperms and their insect herbivores. AB - BACKGROUND: The great diversity of the "Phytophaga" (weevils, longhorn beetles and leaf beetles) has been attributed to their co-radiation with the angiosperms based on matching age estimates for both groups, but phylogenetic information and molecular clock calibrations remain insufficient for this conclusion. METHODOLOGY: A phylogenetic analysis of the leaf beetles (Chrysomelidae) was conducted based on three partial ribosomal gene markers (mitochondrial rrnL, nuclear small and large subunit rRNA) including over 3000 bp for 167 taxa representing most major chrysomelid lineages and outgroups. Molecular clock calibrations and confidence intervals were based on paleontological data from the oldest (K-T boundary) leaf beetle fossil, ancient feeding traces ascribed to hispoid Cassidinae, and the vicariant split of Nearctic and Palearctic members of the Timarchini. PRINCIPAL FINDINGS: The origin of the Chrysomelidae was dated to 73-79 Mya (confidence interval 63-86 Mya), and most subfamilies were post Cretaceous, consistent with the ages of all confirmed body fossils. Two major monocot feeding chrysomelid lineages formed widely separated clades, demonstrating independent colonization of this ancient (early Cretaceous) angiosperm lineage. CONCLUSIONS: Previous calibrations proposing a much older origin of Chrysomelidae were not supported. Therefore, chrysomelid beetles likely radiated long after the origin of their host lineages and their diversification was driven by repeated radiaton on a pre-existing diverse resource, rather than ancient host associations. PMID- 17426810 TI - Haplotype association between haptoglobin (Hp2) and Hp promoter SNP (A-61C) may explain previous controversy of haptoglobin and malaria protection. AB - BACKGROUND: Malaria is one of the strongest recent selective pressures on the human genome, as evidenced by the high levels of varying haemoglobinopathies in human populations-despite the increased risk of mortality in the homozygous states. Previously, functional polymorphisms of Hp, coded by the co-dominant alleles Hp1 and Hp2, have been variously associated with several infectious diseases, including malaria susceptibility. METHODOLOGY/PRINCIPAL FINDINGS: Risk of a clinical malarial episode over the course of a malarial transmission season was assessed using active surveillance in a cohort of Gambian children aged 10-72 months. We report for the first time that the major haplotype for the A-61C mutant allele in the promoter of haptoglobin (Hp)-an acute phase protein that clears haemoglobin released from haemolysis of red cells-is associated with protection from malarial infection in older children, (children aged >or=36 months, >500 parasites/ul and temperature >37.5 degrees C; OR = 0.42; [95% CI 0.24-0.73] p = 0.002) (lr test for interaction, <36 vs >or=36 months, p = 0.014). Protection was also observed using two other definitions, including temperature >37.5 degrees C, dipstick positive, plus clinical judgement of malaria blinded to dipstick result (all ages, OR = 0.48, [95% CI 0.30-0.78] p = 0.003; >or=36 months, OR = 0.31, [95% CI 0.15-0.62] p = 0.001). A similar level of protection was observed for the known protective genetic variant, sickle cell trait (HbAS). CONCLUSIONS/SIGNIFICANCE: We propose that previous conflicting results between Hp phenotypes/genotypes and malaria susceptibility may be explained by differing prevalence of the A-61C SNP in the populations studied, which we found to be highly associated with the Hp2 allele. We report the -61C allele to be associated with decreased Hp protein levels (independent of Hp phenotype), confirming in vitro studies. Decreased Hp expression may lead to increased oxidant stress and increased red cell turnover, and facilitate the development of acquired immunity, similar to a mechanism suggested for sickle cell trait. PMID- 17426811 TI - Phage lambda CIII: a protease inhibitor regulating the lysis-lysogeny decision. AB - The ATP-dependent protease FtsH (HflB) complexed with HflKC participates in post translational control of the lysis-lysogeny decision of bacteriophage lambda by rapid degradation of lambda CII. Both phage-encoded proteins, the CII transcription activator and the CIII polypeptide, are required for efficient lysogenic response. The conserved CIII is both an inhibitor and substrate of FtsH. Here we show that the protease inhibitor CIII is present as oligomeric amphipathic alpha helical structures and functions as a competitive inhibitor of FtsH by preventing binding of the CII substrate. We identified single alanine substitutions in CIII that abolish its activity. We characterize a dominant negative effect of a CIII mutant. Thus, we suggest that CIII oligomrization is required for its function. Real-time analysis of CII activity demonstrates that the effect of CIII is not seen in the absence of either FtsH or HflKC. When CIII is provided ectopically, CII activity increases linearly as a function of the multiplicity of infection, suggesting that CIII enhances CII stability and the lysogenic response. FtsH function is essential for cellular viability as it regulates the balance in the synthesis of phospholipids and lipopolysaccharides. Genetic experiments confirmed that the CIII bacteriostatic effects are due to inhibition of FtsH. Thus, the early presence of CIII following infection stimulates the lysogenic response, while its degradation at later times ensures the reactivation of FtsH allowing the growth of the established lysogenic cell. PMID- 17426812 TI - Transmission of stress-induced learning impairment and associated brain gene expression from parents to offspring in chickens. AB - BACKGROUND: Stress influences many aspects of animal behaviour and is a major factor driving populations to adapt to changing living conditions, such as during domestication. Stress can affect offspring through non-genetic mechanisms, but recent research indicates that inherited epigenetic modifications of the genome could possibly also be involved. METHODOLOGY/PRINCIPAL FINDINGS: Red junglefowl (RJF, ancestors of modern chickens) and domesticated White Leghorn (WL) chickens were raised in a stressful environment (unpredictable light-dark rhythm) and control animals in similar pens, but on a 12/12 h light-dark rhythm. WL in both treatments had poorer spatial learning ability than RJF, and in both populations, stress caused a reduced ability to solve a spatial learning task. Offspring of stressed WL, but not RJF, raised without parental contact, had a reduced spatial learning ability compared to offspring of non-stressed animals in a similar test as that used for their parents. Offspring of stressed WL were also more competitive and grew faster than offspring of non-stressed parents. Using a whole genome cDNA microarray, we found that in WL, the same changes in hypothalamic gene expression profile caused by stress in the parents were also found in the offspring. In offspring of stressed WL, at least 31 genes were up- or down regulated in the hypothalamus and pituitary compared to offspring of non-stressed parents. CONCLUSIONS/SIGNIFICANCE: Our results suggest that, in WL the gene expression response to stress, as well as some behavioural stress responses, were transmitted across generations. The ability to transmit epigenetic information and behaviour modifications between generations may therefore have been favoured by domestication. The mechanisms involved remain to be investigated; epigenetic modifications could either have been inherited or acquired de novo in the specific egg environment. In both cases, this would offer a novel explanation to rapid evolutionary adaptation of a population. PMID- 17426813 TI - Antibiotic stress, genetic response and altered permeability of E. coli. AB - BACKGROUND: Membrane permeability is the first step involved in resistance of bacteria to an antibiotic. The number and activity of efflux pumps and outer membrane proteins that constitute porins play major roles in the definition of intrinsic resistance in Gram-negative bacteria that is altered under antibiotic exposure. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe the genetic regulation of porins and efflux pumps of Escherichia coli during prolonged exposure to increasing concentrations of tetracycline and demonstrate, with the aid of quantitative real-time reverse transcriptase-polymerase chain reaction methodology and western blot detection, the sequence order of genetic expression of regulatory genes, their relationship to each other, and the ensuing increased activity of genes that code for transporter proteins of efflux pumps and down regulation of porin expression. CONCLUSIONS/SIGNIFICANCE: This study demonstrates that, in addition to the transcriptional regulation of genes coding for membrane proteins, the post-translational regulation of proteins involved in the permeability of Gram-negative bacteria also plays a major role in the physiological adaptation to antibiotic exposure. A model is presented that summarizes events during the physiological adaptation of E. coli to tetracycline exposure. PMID- 17426814 TI - Human GLI3 intragenic conserved non-coding sequences are tissue-specific enhancers. AB - The zinc-finger transcription factor GLI3 is a key regulator of development, acting as a primary transducer of Sonic hedgehog (SHH) signaling in a combinatorial context dependent fashion controlling multiple patterning steps in different tissues/organs. A tight temporal and spatial control of gene expression is indispensable, however, cis-acting sequence elements regulating GLI3 expression have not yet been reported. We show that 11 ancient genomic DNA signatures, conserved from the pufferfish Takifugu (Fugu) rubripes to man, are distributed throughout the introns of human GLI3. They map within larger conserved non-coding elements (CNEs) that are found in the tetrapod lineage. Full length CNEs transiently transfected into human cell cultures acted as cell type specific enhancers of gene transcription. The regulatory potential of these elements is conserved and was exploited to direct tissue specific expression of a reporter gene in zebrafish embryos. Assays of deletion constructs revealed that the human-Fugu conserved sequences within the GLI3 intronic CNEs were essential but not sufficient for full-scale transcriptional activation. The enhancer activity of the CNEs is determined by a combinatorial effect of a core sequence conserved between human and teleosts (Fugu) and flanking tetrapod-specific sequences, suggesting that successive clustering of sequences with regulatory potential around an ancient, highly conserved nucleus might be a possible mechanism for the evolution of cis-acting regulatory elements. PMID- 17426815 TI - Evaluation of diagnostic accuracy, feasibility and client preference for rapid oral fluid-based diagnosis of HIV infection in rural India. AB - BACKGROUND: Oral fluid-based rapid tests are promising for improving HIV diagnosis and screening. However, recent reports from the United States of false positive results with the oral OraQuick ADVANCE HIV1/2 test have raised concerns about their performance in routine practice. We report a field evaluation of the diagnostic accuracy, client preference, and feasibility for the oral fluid-based OraQuick Rapid HIV1/2 test in a rural hospital in India. METHODOLOGY/PRINCIPAL FINDINGS: A cross-sectional, hospital-based study was conducted in 450 consenting participants with suspected HIV infection in rural India. The objectives were to evaluate performance, client preference and feasibility of the OraQuick Rapid HIV 1/2 tests. Two Oraquick Rapid HIV1/2 tests (oral fluid and finger stick) were administered in parallel with confirmatory ELISA/Western Blot (reference standard). Pre- and post-test counseling and face to face interviews were conducted to determine client preference. Of the 450 participants, 146 were deemed to be HIV sero-positive using the reference standard (seropositivity rate of 32% (95% confidence interval [CI] 28%, 37%)). The OraQuick test on oral fluid specimens had better performance with a sensitivity of 100% (95% CI 98, 100) and a specificity of 100% (95% CI 99, 100), as compared to the OraQuick test on finger stick specimens with a sensitivity of 100% (95% CI 98, 100), and a specificity of 99.7% (95% CI 98.4, 99.9). The OraQuick oral fluid-based test was preferred by 87% of the participants for first time testing and 60% of the participants for repeat testing. CONCLUSION/SIGNIFICANCE: In a rural Indian hospital setting, the OraQuick Rapid- HIV1/2 test was found to be highly accurate. The oral fluid-based test performed marginally better than the finger stick test. The oral OraQuick test was highly preferred by participants. In the context of global efforts to scale-up HIV testing, our data suggest that oral fluid-based rapid HIV testing may work well in rural, resource-limited settings. PMID- 17426816 TI - The temperature-sensitive role of Cryptococcus neoformans ROM2 in cell morphogenesis. AB - ROM2 is associated with Cryptococcus neoformans virulence. We examined additional roles of ROM2 in C. neoformans and found that ROM2 plays a role in several cell functions specifically at high temperature conditions. Morphologically rom2 mutant cells demonstrated a "tear"-like shape and clustered together. A sub population of cells had a hyperelongated phenotype at restrictive growth conditions. Altered morphology was associated with defects in actin that was concentrated at the cell periphery and with abnormalities in microtubule organization. Interestingly, the ROM2 associated defects in cell morphology, location of nuclei, and actin and microtubule organization were not observed in cells grown at temperatures below 37 degrees C. These results indicate that in C. neoformans, ROM2 is important at restrictive temperature conditions and is involved in several cell maintenance functions. PMID- 17426817 TI - An auditory neural correlate suggests a mechanism underlying holistic pitch perception. AB - Current theories of auditory pitch perception propose that cochlear place (spectral) and activity timing pattern (temporal) information are somehow combined within the brain to produce holistic pitch percepts, yet the neural mechanisms for integrating these two kinds of information remain obscure. To examine this process in more detail, stimuli made up of three pure tones whose components are individually resolved by the peripheral auditory system, but that nonetheless elicit a holistic, "missing fundamental" pitch percept, were played to human listeners. A technique was used to separate neural timing activity related to individual components of the tone complexes from timing activity related to an emergent feature of the complex (the envelope), and the region of the tonotopic map where information could originate from was simultaneously restricted by masking noise. Pitch percepts were mirrored to a very high degree by a simple combination of component-related and envelope-related neural responses with similar timing that originate within higher-frequency regions of the tonotopic map where stimulus components interact. These results suggest a coding scheme for holistic pitches whereby limited regions of the tonotopic map (spectral places) carrying envelope- and component-related activity with similar timing patterns selectively provide a key source of neural pitch information. A similar mechanism of integration between local and emergent object properties may contribute to holistic percepts in a variety of sensory systems. PMID- 17426819 TI - Mature Homogeneous Erythropoietin-Level Building Blocks by Chemical Synthesis: The EPO 114-166 Glycopeptide Domain, Presenting the O-Linked Glycophorin. AB - A synthesis of EPO 114-166 glycopeptide (1), presenting the O-linked glycophorin of erythropoietin, is described. PMID- 17426820 TI - Social support following perinatal loss. AB - The purpose of this project was to examine parents' descriptions of the ways family and friends supported them after they had experienced a perinatal loss. For this project, a secondary analysis of data from two phenomenological studies on perinatal loss was performed. A combined total of 62 interview transcripts from 22 mothers and 9 fathers were examined. Data analysis included identifying all statements in the interview transcripts that pertained to the ways that family and friends supported parents. The modes of supportive behavior (emotional, advice/feedback, practical, financial, and socializing) in Vaux's theory of social support served as a useful framework for presenting the findings. Parents received emotional support most frequently. Findings from the current study provide data for health care professionals to use to provide guidance to family and friends of bereaved parents. PMID- 17426818 TI - Differential cerebral cortex transcriptomes of baboon neonates consuming moderate and high docosahexaenoic acid formulas. AB - BACKGROUND: Docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (ARA, 20:4n 6) are the major long chain polyunsaturated fatty acids (LCPUFA) of the central nervous system (CNS). These nutrients are present in most infant formulas at modest levels, intended to support visual and neural development. There are no investigations in primates of the biological consequences of dietary DHA at levels above those present in formulas but within normal breastmilk levels. METHODS AND FINDINGS: Twelve baboons were divided into three formula groups: Control, with no DHA-ARA; "L", LCPUFA, with 0.33%DHA-0.67%ARA; "L3", LCPUFA, with 1.00%DHA-0.67%ARA. All the samples are from the precentral gyrus of cerebral cortex brain regions. At 12 weeks of age, changes in gene expression were detected in 1,108 of 54,000 probe sets (2.05%), with most showing <2-fold change. Gene ontology analysis assigns them to diverse biological functions, notably lipid metabolism and transport, G-protein and signal transduction, development, visual perception, cytoskeleton, peptidases, stress response, transcription regulation, and 400 transcripts having no defined function. PLA2G6, a phospholipase recently associated with infantile neuroaxonal dystrophy, was downregulated in both LCPUFA groups. ELOVL5, a PUFA elongase, was the only LCPUFA biosynthetic enzyme that was differentially expressed. Mitochondrial fatty acid carrier, CPT2, was among several genes associated with mitochondrial fatty acid oxidation to be downregulated by high DHA, while the mitochondrial proton carrier, UCP2, was upregulated. TIMM8A, also known as deafness/dystonia peptide 1, was among several differentially expressed neural development genes. LUM and TIMP3, associated with corneal structure and age-related macular degeneration, respectively, were among visual perception genes influenced by LCPUFA. TIA1, a silencer of COX2 gene translation, is upregulated by high DHA. Ingenuity pathway analysis identified a highly significant nervous system network, with epidermal growth factor receptor (EGFR) as the outstanding interaction partner. CONCLUSIONS: These data indicate that LCPUFA concentrations within the normal range of human breastmilk induce global changes in gene expression across a wide array of processes, in addition to changes in visual and neural function normally associated with formula LCPUFA. PMID- 17426828 TI - Cholinergic sensory inputs to command neurons in edible snail. AB - We studied cholinergic component of visceral sensory input to defensive behavior command neurons in edible snail. Nicotinic receptor antagonist tubocurarine and muscarinic receptor antagonist atropine reversibly decreased the amplitude of the total excitatory postsynaptic potential induced by electrostimulation of the peripheral region in the mechanosensory receptor field of command neurons on the surface of internal organs. Our results indicate that acetylcholine is involved in sensory signal transduction from the visceral sac to command neurons of snail parietal ganglia. The subsynaptic membrane of visceral synaptic input contains nicotinic and muscarinic receptors. PMID- 17426829 TI - The role of protective effects of proline-containing peptides (PGP, PG, and GP) in contractile dysfunction of mesenteric lymphatic vessels in rats with experimental acute peritonitis. AB - The development of acute peritonitis in rats induced by intraperitoneal injection of thioglycollate was accompanied by a decrease in contractile function of mesenteric lymphatic vessels and impaired response to norepinephrine. Administration of proline-containing peptides after induction of inflammation significantly decreased the severity of these disorders. Our results attest to the possibility of using peptides for the correction of mesenteric microcirculatory disturbances during inflammation. PMID- 17426830 TI - Metabolic activity of cells in brain cortex after alcohol intoxication and correction of changes with Dolivin. AB - Metabolic processes in the brain cortex of albino rats with prenatal and postnatal alcohol intoxication were studied by the method of electron paramagnetic resonance. Accumulation of superoxide radical-generating agents, inactivation of superoxide dismutase, and increase in nitric oxide concentration were detected. Activation of free radical processes was associated with accumulation of Mn(2+)and Fe(2+). The parameters of electron paramagnetic resonance returned to normal after Dolivin treatment. These changes were accompanied by a decrease in the intensity of oxidative metabolism. PMID- 17426831 TI - Selective blockade of central m1 muscarinic cholinergic receptors with pirenzepine impairs cardiovascular and respiratory function in rats with acute hemorrhage. AB - Ultrasound studies showed that selective antagonist of central M1 muscarinic cholinergic receptors pirenzepine (50 mg/kg intravenously) causes transitory hypotension and respiratory depression in anesthetized intact rats. The M1 receptor antagonist had no effect on cardiac output and portal blood flow. Pretreatment with pirenzepine increased the sensitivity of rats with acute massive hemorrhage to circulatory hypoxia. After blockade of central M1 muscarinic cholinergic receptors, the posthemorrhagic period was characterized by primary decompensation of blood pressure, portal blood flow, and respiration and development of low cardiac output syndrome. The animals died over the first minutes after bleeding arrest. Our results indicate that central M1 muscarinic cholinergic receptors act as shock-limiting cholinergic structures under conditions of posthemorrhagic changes in systemic and portal blood flow, as well as during respiratory dysfunction. PMID- 17426832 TI - Antioxidant potential of tear fluid and blood plasma in rabbits with grade III chemical eye burn at various stages of burn disease. AB - The antioxidant potential of tear fluid and plasma in rabbits with chemical eye burn of grade III was measured at various stages of burn disease. Eye burns were accompanied by depletion of the antioxidant system in tear fluid and plasma and impairment of antioxidant defense in the whole organism. These changes contribute to activation of free radical lipid peroxidation. The pathological changes in the organism and eye were most pronounced at the stage of trophic disturbances. PMID- 17426833 TI - Effect of parathyroidectomy on fluctuations in calcium transport between the blood and mineralized tissues of rats. AB - Parathyroidectomy increases the degree of(45)Ca fluctuations between the blood and mineralized tissues (bones and teeth) in rats, which is associated with a decrease in 14C-glycine incorporation into bone proteins. Disturbances in Ca(2+)metabolism and transport during hyperparathyroidism are partly prevented by parathyroidectomy. It is mainly related to variations in the interaction of parathyroid hormone, calcitonin, 1.25(OH)2D3, and other bioactive substances, but not to initiation of mineralization with protein matrixes. PMID- 17426834 TI - Evaluation of the resistance of blood plasma to oxidative stress by oxidizability of proteins and lipids during metal-catalyzed oxidation. AB - A new approach for the evaluation of oxidizability of proteins and lipids in the same sample of blood plasma was proposed. We tested a method for evaluation of metal-catalyzed oxidation of fibrinogen by the formation of bityrosine cross links during oxidation detected by the increase in fluorescence at 415 nm. A correlation was revealed between parameters of oxidizability estimated by this marker and carbonyl derivatives (dinitrophenylhydrazine method). Oxidizability of total proteins from whole plasma was compared with oxidizability of plasma lipids (marker malonic dialdehyde). Study of these parameters in patients with coronary heart disease showed that the proposed experimental approach allows us to divide the sample into several subgroups differing in the resistance to oxidative stress. These data can be used for diagnostic and prognostic purposes. PMID- 17426835 TI - Urokinase induces ROS production in vascular smooth muscle cells. AB - Urokinase stimulates the production of superoxide radical in cultured aortal smooth muscle cells simultaneously with activation of the expression of NAD(F)H oxidases nox1, nox4, and phox22. Antioxidant ebselen abolishes the stimulating effect of urokinase on smooth muscle cell proliferation. The data showed that urokinase can potentiate oxidative stress in the arterial wall and can play an important role in the development of adverse arterial remodeling. PMID- 17426836 TI - Oxidized forms of fibrinogen induce expression of cell adhesion molecules by cultured endothelial cells from human blood vessels. AB - Oxidized forms of fibrinogen similarly to initial non-oxidized fibrinogen induced expression of P-selectin and ICAM-1 cell adhesion molecules in the cultured endothelial cells derived from human umbilical vein. The effect of oxidized fibrinogen on the expression of adhesion molecules was more pronounced. These data attest to more active participation of oxidized forms of fibrinogen into inflammation in the vascular wall, the first stage of atherogenesis. PMID- 17426837 TI - Blood serum DNA in patients with rheumatoid arthritis is considerably enriched with fragments of ribosomal repeats containing immunostimulatory CpG-motifs. AB - We previously hypothesized that the sequence of transcribed region of human ribosomal repeats is selectively accumulated in circulating extracellular DNA due to its increased resistance to double-strand breaks caused by accumulation of single-chain breaks produced by nucleases. The contents of rDNA in blood serum DNA and in DNA from leukocytic nuclei both in healthy donors and in patients with rheumatoid arthritis were compared using dot hybridization method. By the content of non-methylated CpG-repeats, transcribed region of rDNA is identical to bacterial DNA, which is characterized by potent immunostimulatory effect. The transcribed region of rDNA (13.3 kb) contains more than 200 CpG-motifs capable of interacting with TLR9 receptors, which are the mediators of the cell immune response to the action of CpG-rich DNA fragments. The data suggest that DNA from dead cells circulating in the peripheral blood is enriched with sequences possessing potent immunostimulatory properties. PMID- 17426838 TI - Various peptides as substrates for aminopeptidases from salmon eggs during embryogenesis. AB - Seven p-nitroaniline-modified peptides were used as substrates for studies of aminopeptidase and peptidase activities in eggs of lake salmon Salmo salar Sebago at different stages of embryogenesis. The formation of p-nitroaniline during peptide hydrolysis was measured in the incubation medium and in maturing eggs. Activities of all studied aminopeptidases and peptidases increased during the development of fertilized eggs and sharply increased before hatching. L-alanin-p NA, L-pro-p-Na, L-Arg-p-Na, and L-Phe-p-NA were hydrolyzed most of all, while N Glt-L-Phe-p-NA and Z-Gly-Pro-p-NA least of all. PMID- 17426839 TI - Specific activities of poetam preparation (superlow-doses of antibodies to erythropoietin) and recombinant erythropoietin. AB - We compared the capacity of superlow-dose of antibodies to erythropoietin (Poetam) and recombinant erythropoietin (Recormon) to stimulate the recovery of adriamycin-suppressed erythropoiesis in mice. Both preparations exhibited high erythron activation capacity and considerably increased the content of erythrocytes and reticulocytes in the peripheral blood and content of erythrokaryocytes and erythroid precursors in the hemopoietic tissue of experimental animals. The effect of Recormon manifested immediately after injection, while the effect of Poetam was somewhat delayed, but more lasting (due to activation of host erythropoietin system). PMID- 17426840 TI - Inhibition of cytochrome P-450 with 2-diethylamino-ethyl-2,2 diphenylpropylacetate (SKF-525A) reduces immunotoxicity of chlorinated carbohydrates. AB - Experiments on outbred albino rats showed that single intraperitoneal injection of cytochrome P-450 inhibitor 2-diethylaminoethyl-2,2-diphenylpropylacetate (SKF 525A) in a dose of 50 mg/kg before acute poisoning with 1,2-dichloroethane and trichloroethane in a dose of 1.0 LD(50), metabolized in the body to compounds with higher toxicity (the phenomenon of "lethal synthesis") reduced their immunotoxicity by decreasing the formation of their biotransformation products. PMID- 17426841 TI - Systemic effects of energy metabolism regulator amber-anti-tox during vibration induced dysregulation. AB - Energy metabolism regulator Amber-anti-tox had a systemic effect under experimental conditions of vibration-induced dysregulation. Whole-body vibration was accompanied by nonlinear changes in energy metabolism in the heart, liver, kidneys, and lymphocytes and impairment of intra-systemic inter-organ relationships between mitochondria. Amber-anti-tox prevented vibration-induced de energization of mitochondria and contributed to the preservation of multidimensional relationships of energy metabolism in vital internal organs. PMID- 17426842 TI - Generation of reactive oxygen species by umbilical blood cells and immune status of newborns at risk of infectious inflammatory diseases. AB - We carried out a comparative clinical and immunological examination of newborns whose mothers were at risk of infectious inflammatory diseases. Umbilical blood cell phenotype was evaluated by flow cytofluorometry. ROS level was evaluated by chemiluminescence intensity. Spontaneous production of ROS and phagocytic activity of cells in the whole umbilical blood was reduced in newborns born after complicated pregnancy. Low immunoregulatory index indicating changed CD4+/CD8+ ratio and low percentage of natural killer cells were observed in children with manifestations of bacterial infection. ROS production by isolated granulocytes and the effects of PI3K and p38 MAPK (kinases involved in the regulation of activity of NADPH oxidase responsible for the production of ROS) in the risk group infants differed from the corresponding parameters in the control group. The results indicate shifts in the phagocytosis system, immune status, and the receptor-conjugated regulatory systems of ROS generation by granulocytes in newborns at risk of infectious inflammatory diseases. PMID- 17426843 TI - Effect of transplantation of immunocompetent cell on orientation and exploratory behavior and cytokine gene expression in the brain of experimental animals. AB - Intravenous injection of adherent splenocyte fraction from donor (CBAxC57Bl/6)F(1) mice characterized by specific open-field behavior modified this behavior in syngeneic recipient mice. This was paralleled by appropriate changes in the levels of IL-1b and type 1 IL-1 receptor mRNA in the brain cells of recipient mice. Hence, we demonstrated the possibility of directed regulation of orientation and exploratory behavior in mice by transplantation of immunocompetent cells. Mononuclear phagocytes play an important role in this phenomenon. PMID- 17426844 TI - Effect of long-term mental and pain stress on the dynamics of H4 histone acetylation in hippocampal neurons of rats with different levels of nervous system excitability. AB - Priority data on specific effect of long-term mental and pain stress on the dynamics of H4 histone acetylation in the pyramidal neuron nuclei of the hippocampal CA3 field in rats selected by the nervous system excitability were obtained using a comparative genetic method. The congruency of long-term post stress modification of H4 histone acetylation in neurons of rats with high threshold excitability and behavioral changes intrinsic of these rats suggest that increased acetylation of H4 histone together with changes in heterochromatin conformation play a triggering role in long-term modifications of genome expression underlying the pathogenesis of posttraumatic stress disorders and other psychogenias. PMID- 17426845 TI - Walker 256 tumor growth in rats with hereditary defect of vasopressin synthesis. AB - Stable deceleration of Walker 256 tumor growth was detected in Brattleboro rats with vasopressin synthesis defect in comparison with normal WAG rats. In contrast to continuous tumor growth typical of rats, the growth of this tumor in Brattleboro rats was negligible and was observed during the first 15-18 days after transplantation, after which the tumor regressed and disappeared. The effect was age-dependent and was more pronounced in old animals. Repeated injection of Walker 256 cells does not lead to tumor development, which attested to direct involvement of the immune system in the detected phenomenon. PMID- 17426846 TI - Biological activity of hemoglobin-containing complex isolated from blood serum of mice with Ehrlich carcinoma. AB - Injection of hemoglobin-containing complex of serum proteins isolated from animals with Ehrlich carcinoma led to regression of intraperitoneally and intramuscularly transplanted Ehrlich carcinoma in male C57Bl/6 mice. The hemoglobin-containing complex of serum proteins disturbed cycle distribution of Ehrlich carcinoma cells and caused apoptosis of about 34.3% tumor cells. Addition of hemoglobin-containing serum protein complex into Ehrlich carcinoma incubation medium did not lead to the death of tumor cells and even slightly increased their proliferation. PMID- 17426847 TI - Non-starch polysaccharides as correctors in cytostatic therapy of experimental tumors. AB - We studied the effects of low-esterified pectin, calcium pectate, and alginate on the development of transplanted Ehrlich adenocarcinoma and Lewis pulmonary carcinoma and on the efficiency of cyclophosphamide treatment in mice. PMID- 17426848 TI - Geroprotective effect of epithalamine (pineal gland peptide preparation) in elderly subjects with accelerated aging. AB - A 12-year randomized clinical study of epithalamine (pineal gland peptide preparation) was carried out in elderly patients with coronary disease and accelerated aging of the cardiovascular system. Long-term treatment with epithalamine decreased the functional age and degree of cardiovascular aging; exercise tolerance increased. After 12 years the number of elderly subjects dead in the group treated by epithalamine was 28% lower than in the control group, despite the same basic therapy. Cardiovascular mortality was 2-fold lower in patients treated by epithalamine; the incidence of cardiovascular failure and respiratory diseases was 2-fold lower in this group. Long-term treatment with epithalamine was associated with a geroprotective effect on the long-term life prognosis in elderly subjects with accelerated aging. PMID- 17426849 TI - Production of lymphocyte-activating factors by mouse macrophages during aging and under the effect of short peptides. AB - Age-specific characteristics of production of lymphocyte-activating factor by mouse peritoneal macrophages and modulation of this production by short synthetic peptides (Vilon, Epithalon, and Cortagen) were studied. The production of lymphocyte-activating factors by macrophages stimulated with lipopolysaccharides in vitro was lower in old animals. The opposite modulating effects of short peptides on the production of lymphocyte-activating factors by resident and lipopolysaccharide-stimulated macrophages in young and old mice were demonstrated for the first time. This is a possible mechanism of immune system dysfunction during aging, which opens new vistas for its correction with short synthetic peptides. PMID- 17426850 TI - Rheumatoid course of humoral (vascular) rejection after heart allotransplantation. AB - Analysis of planned endomyocardial biopsy specimens of heart allotransplants from 22 recipients revealed signs of humoral type rejection (slight, medium, and severe) presenting as fixation of IgG, IgM, and complement components (C3, C4d) in 61 of 63 sections. Permanent presence of rejection signs attests to rheumatoid course of this process. PMID- 17426851 TI - Functional morphology of chorioallantoic vascular network in chicken. AB - The formation, development, and reduction of the capillary network in the chicken chorioallantoic membrane on days 7-20 of egg incubation were studied by light and electron microscopy with morphometry. Specific features in the architectonics and structure of the mesodermal large vessels and their connection to the suprachoroidal capillaries for provision of adequate gas exchange are shown. PMID- 17426852 TI - Possible reversibility of structural changes in renal vascular bed after correction of experimental coarctation of the aorta. AB - Renal vessels were studied in 22 puppies with induced coarctation of the aorta, 15 with corrected defect, and 12 controls by functional, histological, and morphometrical methods. Constriction of the isthmus of the aorta was associated with decreased pressure of blood flowing to the kidneys and pronounced morphological restructuring of the renal blood system components. Correction of the experimental defect triggers the mechanism of regression of previously induced changes and promotes appreciable recovery of the structure of renal arteries, veins, and glomeruli. PMID- 17426853 TI - Effect of serotonin on gonadal function. AB - The role of endogenous serotonin in the regulation of spermato- and oogenesis was studied in experiments on rabbits. The involvement of the peripheral serotoninergic structures in the studied processes is demonstrated. PMID- 17426854 TI - A novel method for evaluation of ethanol oxidation in living brain. AB - We propose a method for intravital examination of ethanol metabolism in rat brain. Calipsol-anesthetized rats were fixed in stereotaxis and ethanol solution was infused into the lateral cerebral ventricle via an orifice in the skull at a constant rate with a syringe or micro-pump. Elimination of ethanol and production of acetaldehyde (ethanol metabolite) were measured in perfusate samples from the cisterna magna by gas chromatography. The method is highly sensitive, reliable, and reproducible and allows to study the kinetics and enzyme mechanisms of ethanol oxidation in the brain and regulation of this process. PMID- 17426855 TI - Joint spectroscopic and theoretical investigations of transition metal complexes involving non-innocent ligands. AB - A series of transition metal complexes involving non-innocent o-dithiolene and o phenylenediamine ligands has been characterized in detail by various spectroscopic methods like magnetic circular dichroism (MCD), absorption (abs), resonance Raman (rR), electron paramagnetic resonance (EPR), and sulfur K-edge X ray absorption spectroscopies. A computational model for the electronic structure of the complexes is then proposed based on the density functional theory (DFT) or ab-initio methods, which can successfully account for the observed trends in the experimental spectra (MCD, rR, and abs) of the complexes. Based on these studies, the innocent vs non-innocent nature of the ligands in a given transition metal complex is found to be dependent on the position of the central metal ion in the periodic table, its effective nuclear charge in interplay with relativistic effects. PMID- 17426856 TI - Rhodium(III) complexes with a bidentate N-heterocyclic carbene ligand bearing flexible dendritic frameworks. AB - Rh(III) complexes with a bidentate N-heterocyclic carbene ligand bearing flexible dendritic frameworks have been synthesized and fully characterized by X-ray crystallographic analysis, NMR measurements and theoretical calculations. PMID- 17426857 TI - Stepwise nucleophilic substitution of manganocene, syntheses and structures of the dimer [CpMn(hpp)]2 and the unusual manganate cage [LiMn(hpp)3]2 (hppH = 1,3,4,6,7,8-hexahydro-2H-pyrimido[1,2,a]pyrimidine). AB - While the nucleophilic substitution reaction of manganocene [Cp(2)Mn] with Li(hpp) (1 : 1 equivalents) gives the neutral dimer [CpMn(hpp)]2 (1), further substitution of the Cp ligands leads to the unusual dimeric manganate cage compound [LiMn(hpp)3]2 (2), generated by the dimerisation of a simple tris organomanganate monomer. PMID- 17426858 TI - Stereoselective dimerization of racemic C3-symmetric Ti(IV) amine triphenolate complexes. AB - A novel, mononuclear Ti(IV) amine triphenolate complex obtained by reaction of Ti(OiPr)(4) with tris(2-hydroxy-3-phenylbenzyl)amine bearing phenyl ortho substituents affords quantitatively and spontaneously the corresponding heterochiral micro-oxo dinuclear compound whereas an analogous chiral, enantiopure complex maintains its mononuclear structure even in the presence of an excess of water. PMID- 17426859 TI - Allosteric deprogramming of a trinuclear heterometallic helicate. AB - Two multidentate ditopic ligands L1 and L2 which contain both N-donor and crown ether units have been synthesised. The potentially octadentate ligand L1 forms a trinuclear heterometallic double helicate with Cu(I) and Zn(II) ([Zn2Cu(L1)2](5+)), whereas L2 forms a tetranuclear heterometallic double helicate with the same metal ions ([Zn2Cu2(L2)2](6+)). Both species have been characterised by (1)H NMR, ESI-MS and single crystal X-ray crystallography. Reaction of [Zn2Cu2(L2)2](6+) with Ba(2+) results in the coordination of the crown ether units giving the simple barium coordinated species [Zn2Cu2(L2)2Ba2](10+). However, reaction of [Zn2Cu(L1)2](5+) with Ba(2+) deprograms the ligand and results in the formation of a mixture of species. PMID- 17426860 TI - A chiral probe for the detection of Cu(II) by UV, CD and emission spectroscopies. AB - The synthesis and characterization of the new chiral ligand 2, based on a BINOL tren (tren = tris(2-aminoethyl)amine) conjugate, in enantiomerically pure form, are reported. UV-Vis spectrophotometric and spectrofluorimetric studies at variable pH show that the incorporation of a phenolic functionality in close proximity to the polyamino fragment of the molecule causes a dramatic variation of the acid-base and coordination properties of the molecule; the major role of this functionality in determining the selectivity observed in the complexation of Cu(2+), when compared with Zn(2+) and Ni(2+), at pH = 5 in aqueous MeOH, has been clarified, also by comparison with the spectroscopic and coordination properties of model compound 3, bearing protected OMe functionalities. The peculiar absorbance, emission and chiroptical properties of the binaphthyl fragment incorporated in compound 2 can be used for the selective detection of Cu(2+) using three different and potentially orthogonal techniques, namely UV, CD and fluorescence spectroscopies. PMID- 17426861 TI - [n + n]-Heterometallomacrocyclic complexes (n > or = 2) prepared from platinum(II)-centred ditopic 2,2':6',2''-terpyridine ligands: dimensional cataloguing by pulsed-field gradient spin-echo NMR spectroscopy. AB - The reaction of 4'-(2-propyn-1-oxy)-2,2':6',2''-terpyridine (HC[triple bond]CCH2Oterpy) with trans-[PtI2(PR3)2] (R = Et, (n)Bu, Ph) results in the regioselective formation of the metalloditopic ligands trans-[Pt(C[triple bond]CCH2Oterpy)2(PR3)2], crystallographic data for which are presented. Each ditopic ligand reacts with FeCl(2).4H(2)O to give heterometallomacrocycles, the smallest of which is a [2 + 2] macrocycle, confirmed structurally for R = Et. The NMR spectroscopic data confirm the formation of symmetrical species, i.e. macrocyclic and not polymeric species. The distribution of products has been investigated using pulsed-field gradient spin-echo (PGSE) diffusion NMR spectroscopy, and indicates that the kinetic products from the reactions of 1, 2 or 3(L) with iron(II) are [Fe(n)L(n)](2n+) with n = 2, 3 or 4. For L = 1 and 2, these mixtures of products convert in solution to the thermodynamically favoured [Fe(2)L(2)](4+). PMID- 17426862 TI - Dimeric uranyl complexes with bridging perrhenates. AB - The reaction between [UO2(ReO4)2.H(2)O] and two equivalents of either tri-n-butyl phosphine oxide (TBPO) or tri-iso-butyl phosphate (TiBP) results in the formation of [UO2(mu2-ReO4)(ReO4)(TBPO)2]2 (1) and [UO2(mu2-ReO4)(ReO4)(TiBP)2]2 (2) respectively. Both complexes crystallise as two structurally similar centrosymmetric dimers, the cores containing two uranyl moieties linked by bridging perrhenates. Two P=O donor ligands and one monodenatate perrhenate complete the pentagonal bipyramidal coordination sphere at each metal centre. Both complexes have also been characterised in the solid state by vibrational and absorption spectroscopy. Solution spectroscopic characterisation indicates that both perrhenate and phosphine oxide (1) or phosphate (2) remain coordinated, although it is not possible to state conclusively that the dimeric species remain intact. A low resolution structural study of a minor product from the reaction that yielded revealed a monomeric complex with only monodentate perrhenate coordination, [UO2(ReO4)2(H2O)(TiBP)2] (2'). These results represent the first structural evidence for the bridging coordination mode of perrhenate on coordination to an actinide and yields further insight into the possible solvent phase pertechnetate complexes that may exist in PUREX process phosphate rich solvent. PMID- 17426863 TI - Thermodynamic and structural properties of Eu3+ complexes of a new 12-membered tetraaza macrocycle containing pyridine and N-glutaryl groups as pendant arms: characterization of three complexing successive phases. AB - A new polyazamacrocyclic ligand (called pctga) containing pyridine and N-glutaryl arms has been synthesized as a potential agent for MRI (magnetic resonance imaging). Three series of successive complexes formed with Eu(3+) were characterized by at least two of the following methods: potentiometry, EXAFS or luminescence spectrometry. In the immediate complexes [EuH(h)(pctga)(H2O)6](h 3)+**, the metal ion is bound to the oxygen atoms of the three internal carboxylate groups and to six water molecules. As the lanthanide moves into the macrocyclic cavity, these species rapidly evolve into the intermediate metastable complexes [EuH(h)(pctga)(H2O)4](h-3)+*. The formation of two new bonds with the nitrogen atoms of the tetraazamacrocycle decreased the number of coordinated water molecules to four. In the final thermodynamically stable complexes [EuH(h)(pctga)(H2O)(2)](h-3)+, the pctga is bound to the europium(III) in a heptadentate manner, via the four nitrogen atoms of the tetraazamacrocycle and the three oxygen atoms of the internal carboxylate groups. The coordination number of the metal ion is completed to nine with two inner-sphere water molecules. The mean hydration numbers were calculated from the values of the bimolecular quenching constant k(q) of the luminescence species. The thermodynamic parameters corresponding to the protonation constants of the ligand and to the formation constants of the various intermediate and final complexes were determined from potentiometric measurements. They show that the complex species have some specific thermodynamic and structural properties inherent to the N-glutaryl groups and to the pyridine cycle. The insertion of this aromatic substructure rigidifies the ligand and sensibly diminishes the value of the overall formation constant (log beta(110) = 18.66(5)). This whole study allows us to propose a complexation mechanism for the system Eu(3+)/pctga in solution which is a mixture of the ones determined for the ligands dota and tced. PMID- 17426864 TI - Syntheses and structures of pyrazolylmethane complexes of rhenium(III), (IV) and (V). AB - Reaction of [ReOCl3(PPh3)(2)] with HCpz(3) (pz = pyrazole) in dichloromethane leads to the formation of a new Re(iv) complex [ReCl3(HCpz3)]X (X=Cl, [ReO4]) with loss of the rhenium-oxo group. We also report a convenient, high-yield synthetic route to complexes of the type [ReOXn(L)](3-n)+ (X=Cl, Br, n = 2, 3) by the reaction of bis(pyrazolylmethane) and bis(pyrazolylacetate) ligands with [ReOCl3(PPh3)2]. Dinuclear complexes containing the O=Re-O-Re=O group were also isolated and structurally characterised. We have also investigated the reactions of these ligands with diazenide precursors and isolated and characterised complexes of the type [ReClx(N2Ph) (L)(PPh3)] (x = 1,2). The potential applications of these complexes as radiopharmaeuticals is discussed. PMID- 17426865 TI - The synthesis and characterisation of europium terpyridine-N-oxide complexes. AB - Europium complexes of a series of terpyridine-N-oxide ligands have been prepared and structurally characterised by crystallographic studies. While the addition of three equivalents of terpyridine-1-oxide or terpyridine-1,1'-bisoxide results in complexes with nine co-ordinate tricapped trigonal prismatic or monocapped square antiprismatic geometries, respectively, three equivalents of the terpyridine 1,1',1'-trisoxide yields an unexpected 8 co-ordinate geometry. Luminescence studies of the three complexes in acetonitrile show a typical europium emission spectra, dominated by the (5)D(0)-(7)F(2) transition. While no simple trend in the relative quantum yields could be ascertained, the terpyridine-1-oxide complex was observed to have the most intense luminescence for this set of complexes. PMID- 17426866 TI - Steroid analysis in saliva: a noninvasive tool for pediatric research and clinical practice. PMID- 17426867 TI - Breastfeeding and Baby-Friendly Hospital Initiative: more important and with more evidence than ever. PMID- 17426868 TI - Brain arrest, cardiac arrest and uncertainties in defining death. PMID- 17426869 TI - Extrahepatic biliary atresia: current concepts and future directions. AB - OBJECTIVE: To provide an updated review on extrahepatic biliary atresia, focusing mainly on its etiopathogenesis, diagnosis, treatment and prognosis. SOURCES: MEDLINE and PubMed databases were searched using the following keywords: biliary atresia, etiopathogenesis, diagnosis, treatment, prognosis, children. SUMMARY OF THE FINDINGS: Extrahepatic biliary atresia is the main indication for liver transplantation among pediatric patients. As to its etiology, cytomegalovirus, reovirus and rotavirus have been widely investigated as possible triggers of the immunomediated obstruction of the biliary tree. The immune response, especially the predominant TH1 and interferon-gamma responses, genetic susceptibility and disorders related to the embryonic development of the biliary tree can play a role in the etiopathogenesis of extrahepatic biliary atresia. Yet today, portoenterostomy is the only available treatment, with better results when performed in the first 2 months of life. As to prognosis, all untreated children eventually die due to complications resulting from portal hypertension and liver cirrhosis, and most treated children have to undergo liver transplantation. CONCLUSIONS: Extrahepatic biliary atresia is still the major indication for pediatric liver transplantation, and to change this scenario some more light should be shed upon the etiopathogenesis of biliary atresia in different disease phenotypes. Future research into the role of interferon-gamma and of other cytokines is necessary in order to assess whether these aspects should be potential targets for therapeutic intervention. PMID- 17426870 TI - Risk factors for anemia in infants assisted by public health services: the importance of feeding practices and iron supplementation. AB - OBJECTIVE: To investigate risk factors for anemia in infants assisted by public health services. METHODS: In a cross-sectional study carried out in Vicosa, state of Minas Gerais, Brazil, 205 children from 6 to 12 months were evaluated. Socioeconomic, environmental and biological data were collected, as well as information on child's birth, nutritional status, maternal data, child health care practices, feeding practices, and iron supplementation. Diagnosis of anemia was based on hemoglobin levels under 11 g/dL, using a portable Hemocue photometer. To analyze variables associated with anemia, a hierarchical logistic regression model was used. RESULTS: The prevalence of anemia was 57.6%. Family income per capita less than 0.5 minimum wage, frequency of fruit intake less than daily and lack of iron supplementation increased the chance of anemia among infants. CONCLUSION: Adequate health and nutrition support to low income families, promotion of healthy nutritional habits and prescription of iron supplements are of great importance to prevent and manage anemia in infants assisted by public health services. PMID- 17426871 TI - Evolution of body mass index in two historical series of adolescents. AB - OBJECTIVE: To verify the evolution of body mass index (BMI) between two studies of adolescent populations. METHODS: Data on the BMI of 8,020 adolescents aged 10 to 15 years living in the city of Sao Paulo, Brazil, and enrolled on the 2005 study entitled "The nutritional profile of adolescents at public and private schools in Sao Paulo" were compared with data from the 1989 National Nutrition and Health Census (PNSN - Pesquisa Nacional sobre Saude e Nutricao). Binomial testing was used to compare proportions once both data sets had been transformed into percentiles. RESULTS: Comparing the two surveys, significant increases were identified in 85th and 95th percentile BMI values for male adolescents aged 10 to 15 years and for female adolescents aged 10 to 14 years. Analysis of the difference between the 5th and 95th BMI percentiles of the Sao Paulo and PNSN samples indicates that there was probably an increase in the number of adolescents in the higher BMI ranges in Sao Paulo in relation to the PNSN survey. CONCLUSIONS: These results demonstrate a tendency for the adolescents observed mean BMI values to increase during the period between the two surveys, indicating a need for increased monitoring of this measurement as a form of preventing overweight in this population. PMID- 17426872 TI - Impact of obesity on ventilatory function. AB - OBJECTIVE: Although obesity was found to be associated with severe impairment of ventilation, most of the study population has been morbidly obese adults. We aimed to explore the effects of mild obesity on ventilatory function in the pediatric age group. METHODS: In a cross-sectional controlled study, 80 patients (M/F: 35/45), who were evaluated in our outpatient clinic with the complaint of excess body weight, with no history of asthma or other atopic diseases were studied and compared to a control group of 50 normal weight children controlled for age and sex. The mean age of patients was 9.7+/-2.5 years (7 to 15 years). Anthropometric measurements and spirometry were performed in all subjects. Forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1) were used as measures of ventilatory function. RESULTS: There were no significant differences in FEV1%, FVC% and FEV1%/FVC% by study group (p > 0.05). Only three patients had obstructive abnormalities documented on their pulmonary function tests (two had moderately severe and one had mild obstructive abnormalities). No correlation was observed between pulmonary function parameters and anthropometric measurements. CONCLUSION: These data demonstrate that pulmonary function test parameters of the mildly obese children were similar to those of the normal weight children. Anthropometric measurements had no significant effect on spirometric measurements in children as they did on adults. PMID- 17426873 TI - Abdominal circumference as an indicator of clinical and laboratory parameters associated with obesity in children and adolescents: comparison between two reference tables. AB - OBJECTIVE: To evaluate the sensitivity and specificity of two pediatric abdominal circumference reference tables to detect abnormally high body mass index, total cholesterol, fasting blood insulin and leptin levels, and homeostasis model assessment values. METHODS: A total of 624 male and female subjects, with ages ranging from 7 to 18 years, were evaluated. All children were recruited from two public schools. Venous blood samples were collected for determination of fasting plasma insulin, glucose, leptin, and total cholesterol levels. Weight, height and abdominal circumference were assessed according to internationally accepted guidelines. Contingency tables were constructed, comparing the presence or absence of increased abdominal circumference, according to cutoff points established by Taylor et al. and Freedman et al., with the presence or absence of abnormal values in the laboratory tests. RESULTS: Sensitivity values were consistently higher for the table by Taylor et al., whereas the table by Freedman et al. showed greater specificity. Positive predictive values were quite low in general, and were only relevant for body mass index. CONCLUSIONS: Results indicate that the table by Taylor et al. is best for screening purposes, as it identifies individuals at higher risk of presenting abnormal test results. On the other hand, the reference table by Freedman et al. is more suitable for clinical practice, as it could be used to replace laboratory measurements, such as blood insulin or leptin levels, which may not be available at all sites. PMID- 17426874 TI - The effects of fluconazole and cytokines on human mononuclear cells. AB - Candida infections are common infections and fluconazole is one of the most frequently administered antifungal agents in their treatment. The resistance developed against antifungal agents has necessitated the improvement of new treatments. This study focuses on the investigation of the effect of fluconazole and cytokines such as interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), granulocyte-macrophage colony-stimulating factor (GM-CSF) on chemokine production and anticandidal activity of human monocytes. In the study it was observed that GM-CSF caused an increase in candidacidal activity of monocytes. Anticandidal activity of GM-CSF + IFN-gamma combination was not found to be more effective than GM-CSF or IFN-gamma alone. The presence of cytokine and fluconazole caused an increase in the levels of CCL3 and CCL4 chemokines. Accordingly, it was considered that chemokines could contribute to the efficacy of fluconazole in C. albicans infections. Besides, in order to strengthen the immune system some cytokines might be used in addition to antifungal agents for the treatment. PMID- 17426875 TI - Re-mapping the molecular features of the human immunodeficiency virus type 1 and human T-cell lymphotropic virus type 1 Brazilian sequences using a bioinformatics unit established in Salvador, Bahia, Brazil, to give support to the viral epidemiology studies. AB - The analysis of genetic data for human immunodeficiency virus type 1 (HIV-1) and human T-cell lymphotropic virus type 1 (HTLV-1) is essential to improve treatment and public health strategies as well as to select strains for vaccine programs. However, the analysis of large quantities of genetic data requires collaborative efforts in bioinformatics, computer biology, molecular biology, evolution, and medical science. The objective of this study was to review and improve the molecular epidemiology of HIV-1 and HTLV-1 viruses isolated in Brazil using bioinformatic tools available in the Laboratorio Avancado de Saude Publica (Lasp) bioinformatics unit. The analysis of HIV-1 isolates confirmed a heterogeneous distribution of the viral genotypes circulating in the country. The Brazilian HIV 1 epidemic is characterized by the presence of multiple subtypes (B, F1, C) and B/F1 recombinant virus while, on the other hand, most of the HTLV-1 sequences were classified as Transcontinental subgroup of the Cosmopolitan subtype. Despite the high variation among HIV-1 subtypes, protein glycosylation and phosphorylation domains were conserved in the pol, gag, and env genes of the Brazilian HIV-1 strains suggesting constraints in the HIV-1 evolution process. As expected, the functional protein sites were highly conservative in the HTLV-1 env gene sequences. Furthermore, the presence of these functional sites in HIV-1 and HTLV-1 strains could help in the development of vaccines that pre-empt the viral escape process. PMID- 17426876 TI - Trypanosoma cruzi high infectivity in vitro is related to cardiac lesions during long-term infection in Beagle dogs. AB - Trypanosoma cruzi is a hemoflagelate parasite associated with heart dysfunctions causing serious problems in Central and South America. Beagle dogs develop the symptoms of Chagas disease in humans, and could be an important experimental model for better understanding the immunopathogenic mechanisms involved in the chagasic infection. In the present study we investigated the relation among biological factors inherent to the parasite (trypomastigote polymorphism and in vitro infectivity) and immunoglobulin production, inflammation, and fibrosis in the heart of Beagle dogs infected with either T. cruzi Y or Berenice-78 strains. In vitro infectivity of Vero cells as well as the extension of cardiac lesions in infected Beagle was higher for Y strain when compared to Berenice-78 strain. These data suggested that in vitro infectivity assays may correlate with pathogenicity in vivo. In fact, animals infected with Y strain, which shows prevalence of slender forms and high infectivity in vitro, presented cardiomegaly, inflammation, and fibrosis in heart area. Concerning the immunoglobulin production, no statistically significant difference was observed for IgA, IgM or IgG levels among T. cruzi infected animals. However, IgA together IgM levels have shown to be a good marker for the acute phase of Chagas disease. PMID- 17426877 TI - Lutzomyia (Nyssomyia) whitmani s.l . (Antunes & Coutinho, 1939)(Diptera: Psychodidae: Phlebotominae): geographical distribution and the epidemiology of American cutaneous leishmaniasis in Brazil--mini-review. AB - The aim of the present study, in view of the widespread geographical distribution of Lutzomyia (Nyssomyia) whitmani s.l. in Brazil, in close association with the regions of transmission of Leishmania (Viannia) braziliensis and L. (V.) shawi, is to investigate the distribution of this sand fly species and American cutaneous leishmaniasis (ACL) in relationship to vegetation and landscape ecology throughout Brazil. Thematic maps were elaborated by the MapInfo programme, giving information on the spatial distribution of L. whitmani s.l., in accordance with types of vegetation and foci of ACL. With regards to the known areas of transmission of ACL in Brazil, it is notable that L. (N.) whitmani s.l. occurs in most of them, where it has been implicated as a possible vector of L. (V.) braziliensis. The presence of L. whitmani s.l. has been registered in 26 states, the one exception being Santa Catarina; in some states such as Roraima, Acre, Tocantins, and Mato Grosso do Sul this sand fly species has been recorded in a large number of municipalities. L. whitmani s.l. has been found in association with a variety of vegetation types, including the Amazonian forest, savanna ("campos cerrados"), and northeastern savanna ("caatingas nordestinas" or "savana estepica"). PMID- 17426878 TI - Theileria electrophorin.sp., a parasite of the electric eel Electrophorus electricus (Osteichthyes: Cypriniformes: Gymnotidae) from Amazonian Brazil. AB - The name Theileria electrophori n.sp. is proposed for a small parasite described in the erythrocytes of the electric eel, Electrophorus electricus, from Amazonian Brazil. Division of the organism in the erythrocyte produces only four bacilliform daughter cells which become scattered in the host cell, without a cruciform or rosette-shaped disposition. Exoerythrocytic meronts producing a large number of merozoites were encountered in Giemsa-stained impression smears of the internal organs, principally in the liver, and are presumably the source of the intraerythrocytic forms of the parasite. This developmental pattern is characteristic of piroplasms within the family Theileriidae, where the author considers the parasite of E. electricus to most appropriately belong. It effectively distinguishes the organism from the dactylosomatid parasites Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbe, 1894 also found in fishes. This appears to be the second report of Theileria Bettencourt, Franca and Borges, 1907 in a fish. PMID- 17426879 TI - Prevalence of oral hairy leukoplakia and epithelial infection by Epstein-Barr virus in pregnant women and diabetes mellitus patients--cytopathologic and molecular study. AB - Oral hairy leukoplakia (OHL) is generally reported in patients with severe immunosuppression, except for a few cases in individuals with moderate degree of immunodeficiency. It is a white lesion that appears mainly in the lateral border of the tongue, caused by Epstein-Barr virus (EBV). The nuclear changes caused by EBV (Cowdry A inclusion, ground glass and nuclear beading), observed in cytopathology, are specific and enough for the definitive diagnosis of OHL, independent of the identification of the virus. Here we investigated the prevalence of OHL and the presence of EBV-DNA in the lateral borders of the tongue from 90 pregnant women, 90 diabetes mellitus (DM) patients, 30 healthy individuals (negative group) and 30 HIV+ with OHL (positive group). Smears were analyzed by cytopathology and polymerase chain reaction (PCR). A case of subclinical OHL and candidiasis was identificated in a DM patient by cytopathologic analysis. PCR results demonstrated EBV-DNA in 65% of the pregnant women, in 35% of DM patients, and in 20% of the healthy individuals. We concluded that DM patients can develop OHL with a low prevalence. Furthermore, the prevalence of the EBV in lateral border of the tongue is larger in pregnant women than in healthy individuals. PMID- 17426880 TI - Description of Lutzomyia (Lutzomyia) falquetoi sp. nov. (Diptera: Psychodidae, Phlebotominae) a new species from the state of Espirito Santo, Brazil. AB - Lutzomyia (Lutzomyia) falquetoi, sp. nov. (Diptera: Psychodidae, Phlebotominae) is described from the state of Espirito Santo, Brazil. This new species belongs to the series longipalpis and is easily distinguished from the other members of this taxon by the presence of five well-developed spines and a pre-apical spiniform bristle on the gonostyle as well as nine bristles on the basal tuft of the gonocoxite. PMID- 17426881 TI - Genetic relationship of diarrheagenic Escherichia coli pathotypes among the enteropathogenic Escherichia coli O serogroup. AB - The genetic relationship among the Escherichia coli pathotypes was investigated. We used random amplified polymorphic DNA (RAPD) data for constructing a dendrogram of 73 strains of diarrheagenic E. coli. A phylogenetic tree encompassing 15 serotypes from different pathotypes was constructed using multilocus sequence typing data. Phylogram clusters were used for validating RAPD data on the clonality of enteropathogenic E. coli (EPEC) O serogroup strains. Both analyses showed very similar topologies, characterized by the presence of two major groups: group A includes EPEC H6 and H34 strains and group B contains the other EPEC strains plus all serotypes belonging to atypical EPEC, enteroaggregative E. coli (EAEC) and enterohemorrhagic E. coli (EHEC). These results confirm the existence of two evolutionary divergent groups in EPEC: one is genetically and serologically very homogeneous whereas the other harbors EPEC and non-EPEC serotypes. The same situation was found for EAEC and EHEC. PMID- 17426882 TI - Histopathological and ultrastructural aspects of mice lungs experimentally infected with dengue virus serotype 2. AB - Histological and ultrastructural alterations in lung tissue of BALB/c mice infected with dengue virus serotype 2 (non-neuroadapted), by intraperitoneal and intravenous routes were analyzed. Lung tissues were processed following the standard techniques for photonic and electron transmission microscopies. Histopathological and ultrastructural studies showed interstitial pneumonia, characterized by the presence of mononuclear cells. In the mouse model, the dengue virus serotype 2 seems to led to a transient inflammatory process without extensive damage to the interalveolar septa, but caused focal alterations of the blood-exchange barrier. Endothelial cells of blood capillaries exhibited phyllopodia suggesting activation by presence of dengue virus. Morphometrical analysis of mast cells showed an expressive increase of the number of these cells in peribronchiolar spaces and adjacent areas to the interalveolar septa. Alveolar macrophages showed particles dengue virus-like inside rough endoplasmic reticulum and Golgi complex, suggesting viral replication. The tissue alterations observed in our experimental model were similar to the observed in human cases of dengue fever and dengue hemorrhagic fever. Our results show that BALB/c mice are permissive host for dengue virus serotype 2 replication and therefore provides an useful model to study of morphological aspects of dengue virus infection. PMID- 17426883 TI - Observations on Eimeria species of Dasyprocta leporina (Linnaeus, 1758)(Rodentia: Dasyproctidae) from the state of Para, North Brazil. AB - Redescriptions are given of the mature oocysts of Eimeria aguti Carini 1935, E. cotiae Carini, 1935 and E. paraensis Carini, 1935, in the faeces of five specimens of the rodent Dasyprocta leporina (Rodentia: Dasyproctidae) from the state of Para, North Brazil. New information is provided on the sporulation time of these parasites and the prepatent period in experimentally infected D. leporina. Some endogenous stages of E. cotiae are described in the epithelial cells of the ileum, and the absence of any oocysts in the gall-bladder contents of the infected animals indicates that the intestine is also the site of development of E. aguti and E. paraensis. Difficulties in separating E. cotiae and E. paraensis on morphology of the oocysts are discussed. The oocysts of both parasites share many structural features and have a wide size range. It is concluded that although it is at present best to maintain these names, the possibility exists that they were separately given to oocysts of smaller dimensions (E. cotiae) and larger dimensions (E. paraensis) of a single parasite. Location of an endogenous site of development for E. paraensis that is distinctly separate from that of E. cotiae might establish more definitely the separate specific status of the two parasites. PMID- 17426884 TI - Biomphalaria tenagophila potencial vector of Schistosoma mansoni in the Parana River basin (Argentina and Paraguay). AB - Susceptibility and compatibility experiments were carried out with 700 Biomphalaria tenagophila from the Parana River basin exposed to infection with Schistosoma mansoni. Individual infection was performed with 10 miracidia of SJ2 strain from the Paraiba valley (Brazil) originally infective to B. tenagophila. These snails were laboratory-breed progeny of B. tenagophila collected from six localities of Argentina and one from Paraguay. From Argentina: Rincon de Vences (7%) and Posadas (11%) became infected with S. mansoni and the calculation of Frandsen's index (TCP/100) shows that they were Class II poorly compatible. Those snails from Goya (22%), Maloyas (5%), and Beron de Astrada (3%) were Class III compatible to the S. mansoni. None of the 100 snails exposed from Caa-Cati became infected (Class 0 incompatible). Tested samples from Paraguay (Encarnacion) were infected (20%) and compatible (Class III). It was also studied the persistence of the infection in 244 snails of the first generation (F1) of those that were susceptible from three places. It was demonstrated an increment of the susceptibility in the F1 from Maloyas (chi2 = 27.22; p = 0.0001) and Posadas (chi2 = 4.24; p = 0.04). The results point out the possibility that schistosomiasis might be able to spread into the Parana River basin where B. tenagophila exists. PMID- 17426885 TI - Detection of Onchocerca volvulus (Nematoda: Onchocercidae) infection in vectors from Amazonian Brazil following mass Mectizan distribution. AB - Detection of Onchocerca volvulus in Simulium populations is of primary importance in the assessment of the effectiveness of onchocerciasis control programs. In Brazil, the main focus of onchocerciasis is in the Amazon region, in a Yanomami reserve. The main onchocerciasis control strategy in Brazil is the semi-annually mass distribution of the microfilaricide ivermectin. In accordance with the control strategy for the disease, polymerase chain reaction (PCR) was applied in pools of simuliids from the area to detect the helminth infection in the vectors, as recommended by the Onchocerciasis Elimination Program for the Americas and the World Health Organization. Systematic sampling was performed monthly from September 1998 to October 1999, and a total of 4942 blackflies were collected from two sites (2576 from Balawau and 2366 from Toototobi). The molecular methodology was found to be highly sensitive and specific for the detection of infected and/or infective blackflies in pools of 50 blackflies. The results from the material collected under field conditions showed that after the sixth cycle of distribution of ivermectin, the prevalence of infected blackflies with O. volvulus had decreased from 8.6 to 0.3% in Balawau and from 4 to 0.1% in Toototobi. PMID- 17426886 TI - Wild dengue virus types 1, 2 and 3 viremia in rhesus monkeys. AB - Among the flaviviruses, dengue, with its four serotypes, has spread throughout the tropics. The most advanced vaccines developed so far include live attenuated viruses, which have been tested in humans but none has been licensed. Preclinical testing of dengue vaccine candidates is performed initially in mice and in nonhuman primates. In the latter the main criteria used to assay protection are neutralizing antibodies elicited by the vaccine candidate and the magnitude and duration of peripheral viremia upon challenge of previously immunized animals. Towards the identification of wild-type viruses that could be used in challenge experiments a total of 31 rhesus monkeys were inoculated subcutaneously of wild dengue types 1, 2, and 3 viruses. The viremia caused by the different viruses was variable but it was possible to identify dengue viruses useful as challenge strains. PMID- 17426887 TI - Astrovirus infection in children living in the Central West region of Brazil. AB - This study presents data regarding the circulation of astrovirus in Goiania-GO and Brasilia-DF. These viruses were detected in fecal samples from hospitalized children up to five years old with and without acute gastroenteritis. A total of 1244 fecal samples were collected in two periods, 1994 to 1996 (Brasilia) and 1998 to 2002 (Goiania and Brasilia), and were analyzed for viral RNA using the reverse transcription-polymerase chain reaction (RT-PCR). Positivity rates of 4.3 and 0.5% for astrovirus were observed in children with acute gastroenteritis and those without gastroenteritis, respectively. Among children with gastroenteritis no statistically significant difference was seen with regards to viral positivity rates in relation to gender and age. However, a higher incidence rate was observed for children from Brasilia aged 36 months or more. Overall, astroviruses occurred predominantly from September to March in the two cities, suggesting a seasonal pattern for these viruses which coincides with the highest relative air humidity period. The results of this study highlight the importance of astrovirus as an etiologic agent of acute gastroenteritis in children of the Central West region of Brazil. PMID- 17426888 TI - Effects of Brazilian propolis on Leishmania amazonensis. AB - Leishmaniasis, an endemic parasitosis that leads to chronic cutaneous, mucocutaneous or visceral lesions, is part of those diseases, which still requires improved control tools. Propolis has shown activities against different bacteria, fungi, and parasites. In this study we investigated the effect of four ethanolic extracts of typified propolis collected in different Brazilian states, on Leishmania amazonensis performing assays with promastigote forms, extracellular amastigotes, and on infected peritoneal macrophages. Ethanolic extracts of all propolis samples (BRG, BRPG, BRP-1, and BRV) were capable to reduce parasite load as monitored by the percentage of infected macrophages and the number of intracellular parasites. BRV sample called red propolis, collected in the state of Alagoas, and containing high concentration of prenylated and benzophenones compounds, was the most active extract against L. amazonensis. The anti-Leishmania effect of BRV sample was increased in a concentration and time dependent manner. BRV treatment proved to be non-toxic to macrophage cultures. Since BRV extract at the concentration of 25 microg/ml reduced the parasite load of macrophages while presented no direct toxic to promastigotes and extracellular amastigotes, it was suggested that constituents of propolis intensify the mechanism of macrophage activation leading to killing of L. amazonensis. Our results demonstrate, for the first time, that ethanolic extracts of Brazilian propolis reduce L. amazonensis infection in macrophages, and encourage further studies of this natural compound in animal models of leishmaniasis. PMID- 17426889 TI - The number of families of Triatoma dimidiata in a Guatemalan house. AB - Triatoma dimidiata is an important vector of Chagas disease in Guatemala. To help understand the biology and population dynamics of the insect, we estimated the number of full sibling families living in one house. Forty one families with an average size of 2.17 individuals were detected using random amplification of polymorphic DNA-polymerase chain reaction genetic markers. This result suggests high levels of migration of the vector, polyandry, and a significant capability for spreading the disease. PMID- 17426890 TI - Treatment of human acute schistosomiasis with oxamniquine induces an increase in interferon-gamma response to Schistosoma mansoni antigens. AB - Patients with acute schistosomiasis were studied before and after oxamniquine treatment. They had been exposed to cercariae 5 to 9 weeks before, and presented compatible clinical manifestations, eosinophilia, and high levels of total IgE. Interferon-gamma (IFN-gamma) and interleukin-4 were measured by ELISA in whole blood samples under soluble egg antigen or soluble adult worm preparation stimulation. After treatment, the reduction of leukocytosis and eosinophilia were not significant, but total IgE levels decreased significantly, in contrast to IFN gamma levels that were significantly increased. The oxamniquine treatment of acute schistosomiasis patients is followed by an improvement of a Th1 response in vitro. If this response has a protective aspect is unknown, and some investigations need to be realized. PMID- 17426891 TI - Evaluation of cultures of Saccharomyces cerevisae as baits for Triatoma dimidiata and Triatoma pallidipennis. AB - We tested the attraction of Triatoma dimidiata and T. pallidipennis to traps baited with yeast volatiles. Two traps were simultaneously presented in opposite sides of an experimental arena. One trap presented a yeast culture in sucrose solution, while the other contained sucrose solution as control. A first experimental series was done without offering a central refuge for bugs. In a second series, one shelter where the insects could hide was offered and the traps were presented afterwards. In the first series, yeast baited traps attracted significantly more insects than control ones for both species. In the second series, T. pallidipennis was significantly attracted to yeast, whereas T. dimidiata was not attracted. The potential use of yeast baited traps for capturing these vectors of Chagas disease is discussed. PMID- 17426892 TI - What does determine gonad weight in the wild kissing bug Mepraia spinolai. AB - Female fecundity increases with body size in a variety of insects, but it is unknown if this generalization applies for kissing bugs. In this study, we evaluate whether gonad weight in the bloodsucking insect Mepraia spinolai correlates with body size, or determined by nutrition or developmental time. We found that the investment on reproductive tissue correlates positively and significantly with body size and with the amount of ingested blood by female insects along their lifespan. Total molting time did not significantly affect gonad weight. We suggest that under optimal feeding conditions M. spinolai females could express their maximum reproductive potential. PMID- 17426893 TI - New record and larval habitats of Culex eduardoi (Diptera: Culicidae) in an irrigated area of Patagonia, Chubut Province, Argentina. AB - The object of the present work was to identify the larval habitats of Culex eduardoi and to determine the microenvironmental conditions related to their presence in different artificial freshwater environments (temporary, semi permanent, irrigation ditches, and drainage ditches) in tillable areas of Chubut Province, Argentina. This report represents the first record of Cx. eduardoi from this Province and extends its range to latitude 45 degrees S. Immature stages of Cx. eduardoi were found in 8 out of 109 (7.3 %) freshwater habitats and were significantly more prevalent in semi-permanent water bodies. Positive sites had significantly larger surface areas and more vegetation cover than negative sites. PMID- 17426894 TI - Lymnaea cousinni (Gastropoda: Lymnaeidae) as transmitter of fascioliasis. PMID- 17426895 TI - Antimicrobial activity of Uncaria tomentosa against oral human pathogens. AB - Uncaria tomentosa is considered a medicinal plant used over centuries by the peruvian population as an alternative treatment for several diseases. Many microorganisms usually inhabit the human oral cavity and under certain conditions can become etiologic agents of diseases. The aim of the present study was to evaluate the antimicrobial activity of different concentrations of Uncaria tomentosa on different strains of microorganisms isolated from the human oral cavity. Micropulverized Uncaria tomentosa was tested in vitro to determine the minimum inhibitory concentration (MIC) on selected microbial strains. The tested strains were oral clinical isolates of Streptococcus mutans, Staphylococcus spp., Candida albicans, Enterobacteriaceae and Pseudomonas aeruginosa. The tested concentrations of Uncaria tomentosa ranged from 0.25-5% in Mueller-Hinton agar. Three percent Uncaria tomentosa inhibited 8% of Enterobacteriaceae isolates, 52% of S. mutans and 96% of Staphylococcus spp. The tested concentrations did not present inhibitory effect on P. aeruginosa and C. albicans. It could be concluded that micropulverized Uncaria tomentosa presented antimicrobial activity on Enterobacteriaceae, S. mutans and Staphylococcus spp. isolates. PMID- 17426896 TI - A single surgical unit's experience with abdominal tuberculosis in the HIV/AIDS era. AB - INTRODUCTION: Human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) has resulted in a resurgence of abdominal tuberculosis in South Africa, and these patients often present to general surgeons. We describe a single-hospital experience in a region of high HIV prevalence. METHODS: A prospective database of all patients with suspected abdominal tuberculosis was maintained from January 2003 until July 2005. RESULTS: There were 67 patients (20 men, 47 women) with an average age of 32 years (range 27-61 years). The erythrocyte sedimentation rate was universally elevated (105 +/- 23). Altogether, 23 patients were HIV-positive and 7 were HIV-negative. The status was unknown in the remainder. Chest radiographs demonstrated an abnormality in 17 patients (22%). Abdominal ultrasonography was performed in 59 patients and computed tomography in 12. Twelve laparotomies were performed, seven as emergencies. None in the elective laparotomy group died, whereas the mortality rate in the emergency group was 60%. Laparoscopy was insufficient for a variety of reasons. Two patients underwent appendectomy and two excision of a perianal fistula. Two patients underwent biopsy of a palpable subcutaneous node, which confirmed the diagnosis in both cases. A definitive diagnosis was achieved in all 12 patients subjected to laparotomy and at colonoscopic biopsy in 2, lymph node biopsy in 2, appendectomy in 2, perianal fistulectomy in 2, and percutaneous drainage in 2. In the remaining 47 patients the diagnosis was made on the basis of the clinical presentation and radiologic imaging. The patients were commenced on antituberculous therapy. The in-hospital mortality in this group was 10%. Therapy was continued at a centralized tuberculosis facility independent of the hospital. Surgical follow-up was poor, with only five (7%) patients completing the 6-month review at a surgical clinic. CONCLUSIONS: A resurgence in tuberculosis during the HIV era produces a new spectrum of presentations for the surgeon. Emergency surgery is associated with high mortality. Bacterial and histologic evidence of infection are difficult to obtain, and indirect clinical and imaging evidence are used to commence a trial of therapy. A short-term clinical response is regarded as proof of disease. Lack of follow-up means that the efficacy of this strategy is unproven. Health policy changes are needed to enable appropriate surgical follow-up to determine the most effective management algorithm. PMID- 17426897 TI - Transient liver dysfunction after laparoscopic gastrectomy for gastric cancer patients. AB - INTRODUCTION: The effect of laparoscopic surgery under CO2 pneumoperitoneum on liver function is not clear. The aim of this study was to clarify whether laparoscopy-assisted distal gastrectomy (LADG) is associated with changes in liver function compared with open distal gastrectomy (ODG). METHODS: A total of 205 patients who underwent LADG (n = 147) or ODG (n = 58) between January 1994 and April 2004 were included in this study. Liver function tests-aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin, total bilirubin were examined before surgery and at 1, 3, and 7 days after surgery. The postoperative clinical course was compared between the two groups. RESULTS: AST levels on day 1 and ALT levels on days 1 and 3 were significantly higher in the LADG group. Albumin levels showed a marked decrease after operation in both groups, but the level recovered more rapidly in the LADG group than in the ODG group, showing significant differences on days 3 and 7. The total bilirubin levels remained unchanged from baseline. The postoperative complication rate was similar in the two groups, although 3 LADG patients among the 27 patients with liver disease suffered severe enteritis. CONCLUSIONS: Transient liver dysfunction was documented in patients after laparoscopic gastrectomy under CO2 pneumoperitoneum. PMID- 17426899 TI - Germ-line and somatic mutations of the APC gene and/or ss catenin gene in the occurrence of FAP associated thyroid carcinoma. PMID- 17426898 TI - Role of surgeons in clinical trials for thyroid cancer. AB - Properly performed clinical trials provide a foundation for evidence-based medical practice. The surgeon plays a central role in the management of patients with malignant solid tumors, including thyroid cancer, because operative extirpation of the malignancy is the essential first step in effective therapy. This article discusses the role of the surgeon in the clinical research of thyroid cancer and also reviews the important clinical trials that have influenced the treatment of patients with thyroid cancer. Recent discoveries defining the genetic mutations underlying the various types of thyroid cancers have led to the development of targeted therapies. These chemical compounds, which are now being evaluated in clinical trials, hold great promise for the treatment of patients with locally advanced and distant metastatic disease. The surgical investigator also plays an important role in procuring tumor tissue from patients in clinical trials. The molecular analysis of these tissues is of critical importance in selecting specific therapies and predicting patient response and prognosis. PMID- 17426900 TI - Evidence-based endocrine surgery: thyroid cancer. PMID- 17426901 TI - Evidence-based approach to the management of sporadic medullary thyroid carcinoma. AB - Medullary thyroid carcinoma (MTC) is a rare malignancy of the thyroid C cells. It occurs in hereditary (25% of cases) and sporadic (75%) forms. Sporadic MTCs frequently metastasize to cervical lymph nodes. Thorough surgical extirpation of the primary tumor and nodal metastases by compartment-oriented resection has been the mainstay of treatment (level IV evidence). Surgical resection of residual and recurrent disease is effective in reducing calcitonin levels and controlling complications of central neck disease (level IV evidence). Radioactive iodine, external beam radiation therapy, and conventional chemotherapy have not been effective. Newer systemic treatments, with agents that target abnormal RET proteins hold promise and are being tested in clinical trials for patients with metastatic disease. PMID- 17426903 TI - Intrathyroidal differentiated thyroid carcinoma: tumor size-based surgical concepts. AB - INTRODUCTION: Total thyroidectomy (TT) remains the treatment of choice for differentiated thyroid carcinomas (DTCs), but a limited approach can be proposed when tumors are at an early stage and limited to a single lobe. The aim of this study was to analyze the pathologic and clinical aspects of a retrospective series of DTCs in an attempt to determine whether these features might prove useful for limiting the surgical strategy in selected cases. METHODS: From 2000 to 2005, a total of 2798 patients (637 males, 2161 females; mean age 44.6 years) underwent TT for papillary thyroid carcinoma in our department. The histologic features considered were size, histologic subtype and capsule of the tumor, its multifocality/bilaterality, infiltration of the thyroid capsule, and the presence of nodal metastases. RESULTS: Statistical analysis revealed that among tumors < or = 1 cm the presence of the tumor capsule, infiltration of the thyroid capsule, bilaterality, and the presence of node metastases were all significantly lower when the tumor size was < or = 0.5 cm (p < 0.0001). Furthermore, patients with bilateral carcinomas demonstrated a significantly higher presence of a tumor capsule (p = 0.012), infiltration of the thyroid capsule (p < 0.0001), and node metastases (p < 0.0001) and a higher incidence of the "tall-cells" variant (p < 0.0001) when compared to the unilateral population. CONCLUSIONS: Based on these data it is suggested that tumors < or = 1 cm cannot be considered a homogeneous class of DTCs. Nevertheless, in tumors < or = 0.5 cm the absence of a "tall cells" variant and the absence of infiltration of the thyroid capsule might be useful for avoiding an unnecessary completion thyroidectomy after lobectomy. PMID- 17426904 TI - Coagulopathy: its pathophysiology and treatment in the injured patient. AB - Hemorrhage continues to be one of the leading causes of death following trauma. Trauma patients are susceptible to the early development of coagulopathy and the most severely injured patients are coagulopathic on hospital admission. Hypothermia, acidosis, and dilution from standard resuscitation can worsen the presenting coagulopathy and perpetuate bleeding. Early identification of coagulopathy is dependent on clinical awareness and point of care laboratory values. Routinely used laboratory coagulation parameters fail to adequately describe this state. Thrombelastography is a test that can be done at the bedside and uses whole blood to provide a functional evaluation of coagulation. Rapid diagnosis of coagulopathy, followed by prevention or correction of hypothermia and acidosis should be a priority during the initial evaluation and resuscitation. Judicious use of resuscitation fluids and early replacement of coagulation factors will help prevent iatrogenic hemodilution. This review covers the pathophysiology as well as the clinical and laboratory diagnosis of coagulopathy. Prevention and treatment strategies are discussed, including early transfusion of coagulation factors during massive transfusion and the use of recombinant factor VIIa. Damage control resuscitation is briefly discussed, and it involves the combination of hypotensive resuscitation and hemostatic resuscitation. Finally, a description of the use of fresh whole blood in the military setting is included. Its use has been proven to be safe and beneficial in this setting and warrants further investigation as an adjunct to the management of civilian trauma patients. PMID- 17426905 TI - Recurrence pattern of squamous cell carcinoma in the middle thoracic esophagus after modified Ivor-Lewis esophagectomy. AB - BACKGROUND: Despite increasingly radical surgery for esophageal carcinoma, many patients still develop tumor recurrence after operation. This study was designed to evaluate the recurrence pattern of squamous cell carcinoma in the middle thoracic esophagus after modified Ivor-Lewis esophagectomy. METHODS: We retrospectively reviewed data of 196 patients who underwent modified Ivor-Lewis esophagectomy with two-field lymph node dissection from January 1997 to January 2001. Recurrence was classified as locoregional or hematogenous recurrence. Logistic regression analysis was performed to identify risk factors of postoperative recurrence. RESULTS: The overall 3-year and 5-year survival rates in all patients were 53% and 31%, respectively. Recurrence was recognized in 96 patients (48.9%) in the 3 years after operation. The median time to tumor recurrence was 12.2 months. The pattern of recurrence was locoregional in 52 patients (mainly mediastinal in 41, single cervical/supraclavicular in 8), hematogenous in 44 patients (simultaneous locoregional and hematogenous in 10; mainly liver, bone, or lung in 39). The locoregional recurrence rate was significantly lower in patients with postoperative radiotherapy than that in patients without postoperative radiotherapy (p = 0.02). Logistic regression analysis showed that T3 (p = 0.032), N1 (p = 0.003), and postoperative radiotherapy (p = 0.022) were independent risk factors for tumor locoregional recurrence. CONCLUSIONS: About one half of the patients would develop recurrent disease within 3 years after modified Ivor-Lewis esophagectomy with two-field lymph node dissection, and most of them had mediastinal lymph node, liver, bone, or lung metastasis. Postoperative radiotherapy was beneficial in the control of locoregional recurrence. PMID- 17426906 TI - Laparoscopic Nissen-Rossetti fundoplication with routine use of intraoperative endoscopy and manometry: technical aspects of a standardized technique. AB - BACKGROUND: Several different ways of fashioning a total fundoplication lead to different outcomes. This article addresses the technical details of the antireflux technique we adopted without modifications for all patients with GERD beginning in 1972. In particular it aims to discuss the relation between the mechanism of function of the wrap and the physiology of the esophagus. METHODS: The study population consisted of 380 patients affected by GERD with a 1-year minimum of follow-up who underwent laparoscopic Nissen-Rossetti fundoplication by a single surgeon. RESULTS: No conversion to open surgery and no mortality occurred. Major complications occurred in 4 patients (1.1%). Follow-up (median 83 months; range: 1-13 years) was achieved in 96% of the patients. Ninety-two percent of the patients were satisfied with the results of the procedure and would undergo the same operation again. Postoperative dysphagia occurred in 3.5% of the patients, and recurrent heartburn was observed in 3.8%. CONCLUSIONS: Laparoscopic Nissen-Rossetti fundoplication with the routine use of intraoperative manometry and endoscopy achieved good outcomes and long-term patient satisfaction with few complications and side-effects. Appropriate preoperative investigation and a correct surgical technique are important in securing these results. PMID- 17426908 TI - Determination of pain and response to methylprednisolone in Guillain-Barre syndrome. AB - Pain can be a serious problem in patients with Guillain-Barre syndrome (GBS). Different pain symptoms and the effect of methylprednisolone on pain are evaluated. METHODS: GBS patients were recruited from a randomized placebo controlled study comparing intravenous immunoglobulin (IVIg) + methylprednisolone (500 mg for 5 days) versus IVIg + placebo. Presence and severity of pain were prospectively scored at randomization and after 4 weeks. Efficacy of methylprednisolone was evaluated using endpoints: percentage of patients with pain and percentage of patients improving in pain-severity level. Medical records of the subgroup of patients treated in the Erasmus MC were screened retrospectively for different pain symptoms and course. Pain was scored at different time intervals: within 4 weeks before randomization and 0-2, 2-4, 4-24, 24-52 weeks after randomization. RESULTS: 123 (55%) of 223 patients had pain at randomization. In 70%, pain already started before onset of weakness. Methylprednisolone did not show a positive effect on the presence and reduction of pain. In the subgroup of 39 patients, backache (33%), interscapular (28%), muscle (24%), radicular pain (18%) and painful par-/dysaesthesiae (18%) were most frequently present within the period of 4 weeks before randomization. Twenty-six percent had extreme pain 0-2 weeks after randomization. Most symptoms of pain decreased after this period, but painful par-/dysaesthesiae and muscle pain often remained present during at least 6 months. CONCLUSIONS: Pain frequently occurs, often starts before onset of weakness and may cause severe complaints. Especially painful par-/dysaesthesiae and muscle pain may persist for months. Methylprednisolone seems to have no significant effect on the presence and intensity of pain. PMID- 17426909 TI - Apathy and verbal fluency in STN-stimulated PD patients. An observational follow up study. AB - OBJECTIVE: To evaluate apathy and its relation to verbal fluency tasks in a consecutive series of 19 patients with Parkinson's disease (PD) submitted to deep brain stimulation of the subthalamic nucleus (DBS of STN). METHODS: 19 consecutive PD patients submitted to bilateral DBS of STN were studied for apathy pre-operatively and 17 months after surgery. The PD patients underwent a battery of cognitive tests assessing reasoning, memory and frontal executive functions, including phonemic and categorial fluency tasks. The Beck Depression Inventory (BDI) was used for depression. Apathy was assessed by means of the Apathy Scale (AS). In order to quantify changes among individual patients, the clinical criterion of more or less than 1 SD (standard z-score) was used to register a patient as improved or worsened, respectively. RESULTS: After surgery, apathy scores did not change and mood improved (p < 0.02), while a significant worsening was found in the phonemic fluency (p < 0.001). The percentage of patients with an apathy score above the recommended cut-off value (14) was 42% both before and after DBS of STN. Individual outcomes on the apathy scale (1 SD criterion) evidenced that 53% of the patients remained stable, 16% improved, while 31% worsened. This last percentage reduced to 21% (4/19) when considering only the PD patients with an apathy score > or =14 after surgery. No significant correlation was found between the apathy scores variation and any of the neurological variables considered, and, in particular, no correlation was found between apathy and verbal fluency. CONCLUSIONS: The results of the present study suggest that DBS of STN does not necessarily induce apathy even if individual patients show a moderate post-operative worsening of apathetic symptoms. PMID- 17426910 TI - Lumbar spinal stenosis: assessment of cauda equina involvement by electrophysiological recordings. AB - The objective of this study was to investigate the relationship between electrophysiological recordings and clinical as well as radiological findings in patients suggestive to suffer from a lumbar spinal stenosis (LSS). We hypothesise that the electrophysiological recordings, especially SSEP, indicate a lumbar nerve involvement that is complementary to the neurological examination and can provide confirmatory information in less obvious clinical cases. In a prospective cohort study, 54 patients scheduled for surgery due to LSS were enrolled in an unmasked, uncontrolled trial. All patients were assessed by neurological examination, electrophysiological recordings, and magnetic resonance imaging (MRI) of the lumbar spine. The electrophysiological recordings focused on spinal lumbar nerve involvement. RESULTS: About 88% suffered from a multisegmental LSS and 91% of patients respectively complained of chronic lower back pain and/or leg pain for more than 3 months, combined with a restriction in walking distance. The neurological examination revealed only a few patients with sensory and/or motor deficits while 87% of patients showed pathological electrophysiological recordings (abnormal tibial SSEP in 78% of patients, abnormal H-reflex in 52% of patients). CONCLUSIONS: Whereas the clinical examination, even in severe LSS, showed no specific sensory-motor deficit, the electrophysiological recordings indicated that the majority of patients had a neurogenic disorder within the lumbar spine. By the pattern of bilateral pathological tibial SSEP and pathological reflexes associated with normal peripheral nerve conduction, LSS can be separated from a demyelinating polyneuropathy and mono-radiculopathy. The applied electrophysiological recordings, especially SSEP, can confirm a neurogenic claudication due to cauda equina involvement and help to differentiate neurogenic from vascular claudication or musculo-skeletal disorders of the lower limbs. Therefore, electro-physiological recordings provide additional information to the neurological examination when the clinical relevance of a radiologically suspected LSS needs to be confirmed. PMID- 17426911 TI - Vertebral artery dissection presenting with fifth cervical root (C5) radiculopathy. PMID- 17426912 TI - Vitamin D levels in people with multiple sclerosis and community controls in Tasmania, Australia. AB - BACKGROUND: Adequate 25(OH)D levels are required to prevent adverse effects on bone health. Population-based data on factors associated with 25(OH)D levels of people with MS have been lacking. OBJECTIVES: To examine the prevalence and determinants of vitamin D insufficiency in a population-based sample of MS cases and controls, and to compare 25(OH)D status between MS cases and controls, taking into account case disability. METHODS: We conducted a population based case control study in Tasmania, Australia (latitude 41-43 degrees S) on 136 prevalent cases with MS confirmed by magnetic resonance imaging and 272 community controls, matched on sex and year of birth. Measurements included serum 25(OH)D, sun exposure, skin type, dietary vitamin D intake and disability including EDSS. RESULTS: A high prevalence of vitamin D insufficiency was found in MS cases and controls. Among MS cases, increasing disability was strongly associated with lower levels of 25(OH)D and with reduced sun exposure. Cases with higher disability (EDSS > 3) were more likely to have vitamin D insufficiency than controls (OR = 3.07 (1.37, 6.90) for 25(OH)D 0.10 for cup area (0.15), rim area (0.19), maximum cup depth (0.13), height variation contour (0.11), and RNFL cross-sectional area (0.14). The ICC was good (> or =90%) for all parameters, except for mean RNFL thickness (89%) for HRT, and height variation contour (84%) for RTA. CONCLUSION: The observed differences within the limits of agreement were clinically important. Therefore, the two devices cannot be used interchangeably in clinical practice. PMID- 17426924 TI - Improvement of artemisinin production by chitosan in hairy root cultures of Artemisia annua L. AB - Artemisinin production by hairy roots of Artemisia annua L. was increased 6-fold to 1.8 microg mg(-1) dry wt over 6 days by adding 150 mg chitosan l(-1). The increase was dose-dependent. Similar treatment of hairy roots with methyl jasmonate (0.2 mM) or yeast extract (2 mg ml(-1)) increased artemisinin production to 1.5 and 0.9 microg mg(-1) dry wt, respectively. PMID- 17426925 TI - Use of Pseudomonas mendocina, or recombinant Escherichia coli cells expressing toluene-4-monooxygenase, and a cell-free tyrosinase for the synthesis of 4 fluorocatechol from fluorobenzene. AB - The transformation of fluorobenzene (FB) by whole cell expressing toluene-4 monooxygenase (T4MO) resulted in the formation of various hydroxylated products. The predominant product was either 4-fluorophenol (4FP) or 4-fluorocatechol (4Fcat) depending on the ratio of biocatalyst to substrate concentration. The transformation of 1 mM FB by whole cells (1.5 mg CDW/ml) gave a 52% yield of 4Fcat as a single product. The yield of 4Fcat was improved 1.6-fold (80%) by adding 10 mM ascorbic acid to the biotransformations. A combination of two biocatalysts (whole cells expressing T4MO and cell free mushroom tyrosinase) also resulted in the transformation of FB (5 mM) to higher concentrations of 4Fcat (1.8 mM) compared to a whole cell biotransformation alone. However, mixed products were formed and the yield of 4Fcat from FB was lower using the two-step (tandem) method (27%) compared to the use of whole cells of P. mendocina KR1 alone (80%). PMID- 17426926 TI - Alterations in expression of senescence marker protein-30 gene by 3,3',5-triiodo L-thyronine (T3). AB - Thyroid hormone (T3) is essential for normal development, differentiation, and metabolic balance of the body. A toxic dose of T(3) in animals increases the basal metabolic rate and reactive oxygen species production, resulting more oxidative stress through Ca(2+) influx to cytoplasm. Senescence Marker Protein-30 (SMP30) is preferentially expressed in the liver and protects cells against various injuries by enhancement of Ca(2+) efflux to either extra cellular space or intraorganellar spaces through membrane Ca(2+) pump activity. In this paper we report an alteration in the level of SMP30 gene expression using RT-PCR and western blot analysis in T(3) treated female Wistar rats. The results indicate that there is an induction of SMP30 expression during early hours of T(3 )treatment and it declines in severe hyperthyroidism. Therefore, we speculate that SMP30 is regulated by T(3) and might play a protective role in hyperthyroidism. PMID- 17426927 TI - Saturated fatty acid-mediated endoplasmic reticulum stress and apoptosis are augmented by trans-10, cis-12-conjugated linoleic acid in liver cells. AB - Lipid accumulation in non-adipose tissues leads to cell dysfunction and apoptosis, a phenomenon known as lipotoxicity. Recent evidence suggests that lipotoxicity in hepatocytes involves endoplasmic reticulum (ER) stress and c-Jun NH(2)-terminal kinase-mediated apoptosis. The present study examined (1) the dose response and time course characteristics of fatty acid-mediated ER stress and apoptosis in H4IIE liver cells; (2) whether saturated fatty acid-induced apoptosis involved the ER-associated caspase-12; and (3) whether trans-10, cis-12 conjugated linoleic acid, an inhibitor of stearoyl-CoA desaturase, influenced fatty acid-mediated ER stress and apoptosis. Saturated fatty acids induced ER stress in a dose-dependent manner with a time course that was delayed relative to chemical-induction of ER stress. Saturated fatty acids increased caspase-9 and caspase-3 activity, however increased caspase-12 activity was not observed. Inhibition of stearoyl-CoA desaturase, using conjugated linoleic acid (trans-10, cis-12), augmented saturated fatty acid-induced ER stress and apoptosis. These data suggest that saturated fatty acids induce ER stress and apoptosis at physiologic concentrations and with a relatively rapid time course. It would appear that saturated fatty acid-mediated apoptosis occurs independently of caspase-12 activation. Since conjugated linoleic acid inhibited stearoyl-CoA desaturase activity, it is hypothesized that saturation, per se, plays a role in lipotoxicity in liver cells. PMID- 17426928 TI - Proteomic analysis of MOLT-4 cells treated by valproic acid. AB - The effect of valproic acid (VA) on protein expression in human T-lymphocytic leukemia cells MOLT-4 was studied. VA is an inhibitor of histonedeacetylases and has a potential use as antitumor agent in leukemia treatment. The authors in this work prove that 4 h long incubation with 2 mmol/l VA causes phosphorylation of histone H2A.X and its colocalization with 53BP1 in nuclear foci. Their co localization is typical for DSB signaling machinery. These foci were detected in cells after 4 h exposure without increase of Annexin V positive apoptotic cells. Slight increase in apoptosis (Annexin V positivity) after 24 h is accompanied by more intensive increase in phosphorylation of H2A.X and also by formation of nuclear foci containing gammaH2A.X and 53BP1. Treatment of cells with 2 mmol/l VA resulted in induction of apoptosis affecting about 30% of cells after incubation for 72 h. The changes in protein expression were examined after cell incubation with 2 mmol/l VA for 4 h. Proteins were separated by two-dimensional electrophoresis and quantified using image evaluation system. Those exhibiting significant VA-induced abundance alterations were identified by mass spectrometry. Changes in expression of 22 proteins were detected, of which 15 proteins were down-regulated. Proteomic analysis resulted in successful identification of three proteins involving alfa-tubulin 3, tubulin-specific chaperone and heterogeneous nuclear ribonucloprotein F. Expression of seven proteins was up-regulated, including heterogeneous nuclear ribonucloprotein A/B. Identified proteins are related to microtubular system and hnRNP family. Suppression of microtubular proteins and changes of balance among hnRNPs can contribute to proliferation arrest and apoptosis induction. PMID- 17426929 TI - Angiotensin receptor blockade and angiotensin-converting-enzyme inhibition limit adverse remodeling of infarct zone collagens and global diastolic dysfunction during healing after reperfused ST-elevation myocardial infarction. AB - To determine whether therapy with the angiotensin II type 1 receptor blocker (ARB) candesartan and the comparator angiotensin-converting-enzyme inhibitor (ACEI) enalapril during healing after reperfused ST-elevation myocardial infarction (RSTEMI) limit adverse remodeling of infarct zone (IZ) collagens and left ventricular (LV) diastolic dysfunction, we randomized 24 dogs surviving anterior RSTEMI (90-min coronary occlusion) to placebo, candesartan, and enalapril therapy between day 2 and 42. Six other dogs were sham. We measured regional IZ and non-infarct zone (NIZ) collagens (hydroxyproline; types I/III; cross-linking), transforming growth factor-beta (TGF-beta) and topography at 6 weeks, and hemodynamics, LV diastolic and systolic function, and remodeling over 6 weeks. Compared to sham, placebo-RSTEMI differentially altered regional collagens, with more pronounced increase in TGF-beta, hydroxyproline, and type I, insoluble, and cross-linked collagens in the IZ than NIZ, and increased IZ soluble and type III collagens at 6 weeks, and induced persistent LV filling pressure elevation, diastolic and systolic dysfunction, and LV remodeling over 6 weeks. Compared to placebo-RSTEMI, candesartan and enalapril limited adverse regional collagen remodeling, with normalization of type III, soluble and insoluble collagens and decrease in pyridinoline cross-linking in the IZ at 6 weeks, and attenuation of LV filling pressure, diastolic dysfunction, and remodeling over 6 weeks. The results suggest that candesartan and enalapril during healing after RSTEMI prevent rather than worsen adverse remodeling of IZ collagens and LV diastolic dysfunction, supporting the clinical use of ARBs and ACEIs during subacute RSTEMI. PMID- 17426930 TI - Typical phenotypic spectrum of velocardiofacial syndrome occurs independently of deletion size in chromosome 22q11.2. AB - Velocardiofacial syndrome (VCFS) is a relatively common developmental disorder characterized by craniofacial anomalies and conotruncal heart defects. Many VCFS patients present hemizygous deletions on part of chromosome 22q11.2; suggestive that haploinsufficiency in this region is responsible for this etiology. Most 22q11.2 deletions occur sporadically, although in some cases the deletion may be transmitted. A total of 29 VCFS patients and their parents were genotyped using six consecutive polymorphic markers (STS) of the chromosome 22q11.2: D22S420, D22S941, D22S264, D22S306, D22S425, and D22S257. The results revealed that 72% (21/29) of the patients harbored a deletion involving the polymorphic markers D22S420, D22S941, and/or D22S264. Haplotype analysis showed that among the patients studied, the deletions were either of maternal or paternal origin. Our findings demonstrated that independently of their size, any deletion occurring in the VCFS critical region is enough to confer the patient phenotype. PMID- 17426931 TI - Glyceraldehyde-3-phosphate dehydrogenase exerts different biologic activities in apoptotic and proliferating hepatocytes according to its subcellular localization. AB - Recent evidences indicate new roles for the glycolytic protein glyceraldehyde-3 phosphate dehydrogenase (GAPDH) in essential mammalian cell processes, such as apoptosis and proliferation. To clarify the involvement of this protein in growth and programmed cell death in the liver, cell models of hepatocytes in culture were used to study GAPDH expression, localization and enzymatic activity in hepatocyte proliferation and apoptosis. GAPDH expression in cell compartments was studied by Western blot. Nuclear expression of GAPDH increased in apoptosis, and cytoplasmic expression was elevated in apoptosis and proliferation. Subcellular localization was determined by GAPDH immunostaining and confocal microscopic analysis. Quiescent and proliferating hepatocytes showed cytoplasmic GAPDH, while apoptotic cells showed cytoplasmic but also some nuclear staining. The glycolytic activity of GAPDH was studied in nuclear and cytoplasmic cell compartments. GAPDH enzymatic activity increased in the nucleus of apoptotic cells and in cytoplasms of apoptotic and proliferating hepatocytes. Our observations indicate that during hepatocyte apoptosis GAPDH translocates to the nucleus, maintaining in part its dehydrogenase activity, and suggest that this translocation may play a role in programmed hepatocyte death. GAPDH over-expression and the increased enzymatic activity in proliferating cells, with preservation of its cytoplasmic localization, would occur in response to the elevated energy requirements of dividing hepatocytes. In conclusion, GAPDH plays different roles or biological activities in proliferating and apoptotic hepatocytes, according to its subcellular localization. PMID- 17426932 TI - [Pathobiology of acute myeloid leukemia]. AB - Acute myeloid leukemia (AML) arises from the clonal expansion of primitive myeloid precursor cells. A series of genetic alterations leads to a perturbation of normal developmental programs affecting growth, maturation and differentiation of hematopoietic cells. As a consequence, immature leukemic cells that have the ability to divide and proliferate, but lack normal differentiation mechanisms, accumulate in the bone marrow. This, in turn, leads to a severe impairment of normal hematopoiesis. Over the last several years, a number of clinical and basic research studies have elucidated important pathogenetic mechanisms leading to the initiation of AML. The identification of numerous chromosomal aberrations and mutations specific for AML has deepened our insights into the biology of AML and has allowed to improve our diagnostic tools, the definition of prognostic subgroups and therapeutic concepts. PMID- 17426933 TI - [Diagnostics, classification and prognostic criteria of acute myeloid leukemia]. AB - DIAGNOSTICS: The continuously growing knowledge about criteria important for biology, pathogenesis, prognosis and treatment of acute myeloid leukemia (AML) necessitates a broad spectrum of diagnostic methods for first diagnosis and for the further course of the disease. Relevant diagnostic techniques (cytomorphology with cytochemistry, immunophenotyping, cytogenetics and molecular genetics, DNA array) are described - with a focus on their mode of operation as well on their clinical significance. Due to the high clinical relevance and growing complexity, AML diagnostics should be performed in specialized laboratories. CLASSIFICATION: Compared to the FAB classification which is based primarily on morphological criteria, the classification recommended in 2001 by the WHO additionally takes cytogenetics, molecular genetics and further clinical factors into consideration. Both classifications are described. PROGNOSTIC CRITERIA: A wide range of prognostic criteria of AML is discussed on the basis of currently available clinical data. The most important criteria are the karyotype of the leukemic clone and the patient's age. PMID- 17426934 TI - [Current therapeutic strategies in the management of acute myeloid leukemia]. AB - Acute myeloid leukemia (AML) is a rare disease of the hematopoietic stem cell leading to uncontrolled proliferation of immature progenitor cells. This results in a replacement of healthy hematopoiesis and in pancytopenia with corresponding symptoms (anemia, thrombo- and granulocytopenia). Diagnosis can reliably be confirmed by bone marrow aspiration, which also allows risk stratification by cytogenetic and molecular analysis. Therapy of AML that should preferentially be performed in clinical studies comprises induction therapy for achievement of complete cytomorphological remission (CR) and postremission strategies consisting of consolidation and maintenance therapy for eradication of residual blasts. The backbone of polychemotherapy is cytarabine and anthracyclines. Different regimens exist that achieve CR rates of 60-80%. As a consequence, pancytopenias up to 6 weeks will be experienced, that will lead to specific problems such as infections by atypical pathogens. To date, induction therapy will be performed independently of the individual risk constellation (with the exception of acute promyelocytic leukemia); however, postremission therapy is highly dependent on individual risk stratification. Besides conventional strategies, allogeneic stem cell transplantation has to be considered in certain risk groups depending on the availability of a matched donor. Taken together, cure can be achieved in about 40% of patients, however, with large interindividual variability. PMID- 17426935 TI - [Allogeneic stem cell transplantation in the management of acute myeloid leukemia]. AB - Allogeneic stem cell transplantation (SCT) is the most powerful treatment option for acute myeloid leukemia (AML). However, SCT is also complicated by a high risk for treatment-related morbidity and mortality. The antileukemic effect of SCT is based on the radio-/chemotherapy applied for conditioning, as well as on the allogeneic immune reaction, mediated by immunocompetent donor cells, the graft versus-leukemia effect. The latter effect is of particular importance in the context of reduced-intensity conditioning regimens, that have enabled us to offer allogeneic SCT to a by far bigger part of patients suffering from AML. The indication for allogeneic SCT is based on the patient's individual risk profile. Biological and clinical characteristics of the leukemia contribute to this risk profile, as do extraleukemic conditions such as age and comorbidity. Allogeneic SCT represents the standard of care for all patients with AML < 65 years of age, who are beyond first complete remission (CR) or who have failed to respond to induction chemotherapy. In first CR, allogeneic SCT is a standard for patients with unfavorable karyotype disease or other risk factors, whereas for patients without specific risk factors it is just an option, in particular within clinical trials. In patients with a favorable leukemic karyotype, allogeneic SCT is usually not performed in first CR. Future developments in the field include transplant strategies specifically designed for biological AML subgroups, as well as the integration of new drugs into transplant regimens. PMID- 17426936 TI - [Therapy of acute myeloid leukemia (AML) for medically non-fit patients]. AB - Acute myeloid leukemia (AML) has an increasing incidence with higher age, which is about 15 per 100,000 for patients > 65 years. Many older AML patients show functional restrictions and have a high comorbidity status, so that they do not seem to be qualified for a curatively intended chemotherapy. Decisive for the low cure rate of older AML patients are both patient-dependent and disease-dependent reasons such as secondary or therapy-related leukemia or adverse cytogenetics with complex chromosomal abnormalities, which are poor prognostic factors and are responsible for the low probability to achieve long-lasting complete remissions. Prognostic scores are developed for identifying "medically non-fit" patients as objectively as possible. In the future, these patients should not only be offered best supportive care but also well-tolerable concepts of therapy, which are feasible on an ambulatory basis. The main aim for this patient group must be to avoid long hospitalizations and to maintain a high quality of life. PMID- 17426937 TI - Granular dynamics, jamming, rheology and instabilities. PMID- 17426938 TI - Initiation and evolution of current ripples on a flat sand bed under turbulent water flow. AB - We investigate the formation and dynamics of sand ripples under a turbulent water flow. Our experiments were conducted in an open flume with spherical glass beads between 100 and 500 microm in diameter. The flow Reynolds number is of the order of 10,000 and the particle Reynolds number of the order of 1 to 10. We study the development of ripples by measuring their wavelength and amplitude in course of time and investigate the influence of the grain size and the flow properties. In particular, we demonstrate two different regimes according to the grain size. For fine grains, a slow coarsening process (i.e., a logarithmic increase of the wavelength and amplitude) takes place, while for coarser grains, this process occurs at a much faster rate (i.e., with a linear growth) and stops after a finite time. In the later case, a stable pattern is eventually observed. Besides, we carefully analyze the wavelength of ripples in the first stages of the instability as a function of the grain size and the shear velocity of the flow, and compare our results with other available experimental data and with theoretical predictions based on linear stability analyses. PMID- 17426939 TI - Correlations and aggregate statistics in granular packs. AB - We study how the aggregate statistical properties for density fluctuations in granular aggregates scale with the sample size and how such a scaling is associated with the correlations between grains. Correlations are studied both between grain positions and between Voronoi cell volumes, showing distinct behaviors and properties. A non-linear scaling in the aggregate volume fluctuations as function of the sample size is discovered and the connection between such anomalous scaling and correlations is explained. It emerges that volume fluctuations might be described by means of a single universal equation for all samples at all cluster sizes. PMID- 17426940 TI - [Ultrasound during follow-up of carcinoma]. AB - According to the guidelines, ultrasonography (US) is now established as the cross sectional imaging technique of choice in postoperative care of colorectal carcinoma. Although conventional percutaneous US is inferior to computed tomography (CT) and magnetic resonance imaging (MRI) for detecting hepatic metastases, the application of specific contrast media has significantly increased sensitivity and specificity to 87% and 88%, respectively. The combination of US and CT/MRI achieves the highest detection rates. During follow up of rectal carcinoma, in up to 20% of locoregional recurrences are diagnosed solely by endorectal sonography and result in repeat resection with curative intention. In noncolorectal carcinoma, US is recommended in the guidelines for following up hepatocellular carcinoma and malignant thyroid disease, but the available data are insufficient to support those recommendations. PMID- 17426941 TI - [Three-dimensional echocardiography in cardiac surgery. Current status and perspectives]. AB - Three-dimensional (3D) echocardiography is a new imaging technique that can provide useful information about cardiovascular morphology, pathology, and function. Recent refinements in instrumentation, data acquisition, post processing, and computation speed allow 3D echocardiography to play an important role in cardiac imaging. These modalities provide comprehensive information on ventricular and valve morphology and function. Combined with 3D color Doppler sonography, further assessment of valvular function and determination of flow in the left ventricular outflow tract and cross-septal defects are now possible. Three-dimensional color flow imaging also makes echocardiography accurate for assessing the severity of mitral regurgitation. The purpose of this review is to describe technical developments in 3D echocardiography and its clinical application in cardiac surgery. Moreover, based on clinical studies at our centre, we describe the morphology of the mitral valve, its flow pattern, and function of the mitral annulus. PMID- 17426942 TI - [Benign prostate hyperplasia: success and limitations of pharmacological therapy]. AB - A profound knowledge of pathogenesis and natural history enables a differentiated therapy for elderly men with lower urinary tract symptoms due to benign prostatic hyperplasia (BPH). The role of phytotherapy is still controversially discussed and, therefore, not clearly recommended by any BPH-guideline. alpha(1)-blockers are the therapy of choice for symptomatic patients at a low risk of disease progression (prostate volume <30-40 ml). 5alpha-reductase inhibitors (5ARI) reduce the prostate volume by 20-25% and the risk for acute urinary retention/surgery by more than 50% compared to placebo. Combination therapy (alpha(1)-blocker plus 5ARI) is superior to either monotherapy, though this advantage is only demonstrable after a prolonged treatment period (>12 months). PMID- 17426943 TI - [Doping. High-tech cheating in sport]. AB - Today, doping is no longer limited to the classical drugs with well known effects and side effects. Older generation anabolic steroids are used mainly in fitness and recreational sports. In contrast, due to doping tests, substances used in competitive sports include peptide hormones, medications not yet approved, and even specially developed drugs, such as designer steroids. Of the peptide hormones, particularly growth hormones (human growth hormone), erythropoietin and generics, insulin, and presumably insulin-like growth factor 1 are used. Substance groups potentially relevant for doping are selective androgen receptor modulators and gene therapy drugs. For most of these, there is no knowledge about side effects in healthy individuals, and no adequate doping tests. Therefore, anti-doping measures cannot rely solely on the continual improvement of doping analyses, but should include increased measures for doping prevention. Not only sports organizations, but also governmental agencies should be involved in developing and implementing these measures. PMID- 17426944 TI - [Postoperative symptoms and surgical sequelae--their meaning for internists]. PMID- 17426945 TI - [Pneumonias and immunosuppression]. AB - Pneumonias are common, life threatening complications in immunodeficient patients. A rapid, targeted diagnosis and therapy are decisive for the course of the disease. The etiological spectrum is substantially broader than for immunocompetent patients. Important indications are provided by the type and intensity of the immunosuppression, gaps in the prophylactic concept and particular exposures. For diagnostic planning, high resolution computed tomogram of the thorax is necessary. The standard method for isolating the pathogen is flexible bronchoscopy with bronchoalveolar lavage. Indications for invasive bioptic measures depend on the individual situation, the expected spectrum of pathogens as well as risk factors and adverse effects. Non-invasive antigen and PCR tests complement the diagnostic spectrum, especially for difficult to cultivate pathogens such as fungi and Herpes viruses. The selection of the initial, targeted antimicrobial therapy is based on guidelines. The treatment should be followed after obtaining the relevant microbiological data. PMID- 17426946 TI - [Obscure coughing and the feeling of suffocation during sleep. Long-term nocturnal acoustic recording]. AB - A 58-year old male presented with a 6-week history of increasing nocturnal cough, dyspnea and awakening. Physical examination, chest auscultation and lung function in the daytime did not show any pathological findings. Only nocturnal long-term recording of respiratory sounds was helpful: In- and expiratory wheezing, as the acoustic manifestation of airway obstruction, was found throughout the night. As the cause for nocturnal cough and distinct airway obstruction, a pansinusitis with postnasal drip syndrome was diagnosed. PMID- 17426947 TI - [Aortocoronary bypass and cardiac valve surgery]. AB - In Germany, approximately 100,000 patients underwent cardiac surgery in 2005. The most important postoperative problems with prognostic impact for these patients were acute heart failure, acute renal failure, infections, neurological complications, postcardiotomy syndrome, cardiac tamponade and atrial fibrillation. The "EuroSCORE" is a relevant predictor of immediate and long-term outcome in these patients. The patency rates of coronary artery bypass grafts (CABG) are important for the long-term outcome in such surgery. There has been a significant increase in the number of patients aged 80 years and older who are referred for cardiac surgery, which is an outstanding challenge for cardiac surgeons. PMID- 17426948 TI - Stromal-cell regulation of natural killer cell differentiation. AB - Natural killer (NK) cells are bone-marrow-derived lymphocytes that play a crucial role in host defense against some viral and bacterial infections, as well as against tumors. Their phenotypic and functional maturation requires intimate interactions between the bone marrow stroma and committed precursors. In parallel to the identification of several phenotypic and functional stages of NK cell development, recent studies have shed new light on the role of stromal cells in driving functional maturation of NK cells. In this review, we provide an overview of the role of bone marrow microenvironment in NK cell differentiation. PMID- 17426949 TI - Mitochondrial channelopathies in aging. AB - Defects in ion channels (channelopathies) are increasingly found in a large spectrum of human pathologies including aging. Mutations in genes encoding ion channel proteins, which disrupt channel function, are the most commonly identified cause of channelopathies. Mutations in associated proteins, alterations in the expression of ion channels, or changes in the activity of non mutated channel genes or associated proteins can also produce acquired channelopathies. Mitochondria, the powerhouse of the cells, are considered to be the most important cellular organelles to contribute to aging mainly because of their role in the production of reactive oxygen species in the initiation of apoptotic cell remodeling and in efficient ATP synthesis. During the past 50 years, multiple ion channels or transporters have been found in mitochondria, and the relationship between the activity of these channels and cellular aging, as well as the overall cellular biological function, has been intensively studied in a number of cell types and animal models. In this review, we discuss the better characterized mitochondrial ion channels whose dysfunction (mitochondrial channelopathies) may affect or accelerate the aging processes. These channels include the mitochondrial ATP-sensitive potassium channel (mitoK(ATP)), Ca(2+) transporters, voltage-dependent anion channel, and the mitochondrial permeability transition pore (mitoPTP). PMID- 17426950 TI - Laminin isoforms in development and disease. AB - The members of the laminin family of heterotrimers are major constituents of all basement membranes, sheet-like extracellular structures, present in almost all organs. The laminins bind to cell surface receptors and thereby tightly connect the basement membrane to the adjacent cell layer. This provides for the specific basement membrane functions to stabilize cellular structures, to serve as effective physical barriers, and furthermore, to govern cell fate by inducing intracellular signalling cascades. Many different types of diseases involve basement membranes and laminins. Metastasizing solid tumors must pass through basement membranes to reach the vascular system, and various microbes and viruses enter the cells through direct interaction with laminins. Furthermore, whereas mutations in one specific laminin chain lead to a muscular disorder, mutations of other laminin chains cause skin blistering and kidney defects, respectively. This review summarizes recent progress concerning the molecular mechanisms of laminins in development and disease. The current knowledge may lead to clinical treatment of lamininopathies and may include stem-cell approaches as well as gene therapy. PMID- 17426951 TI - [Interactional therapy program for mothers with postpartum mental disorders. First results of a pilot project]. AB - BACKGROUND: The period around delivery frequently causes psychiatric diseases that may disturb maternal competence and influence bonding behaviour with the child. Until now only a few possibilities have existed for inpatient treatment and therapy for these problems. The therapy program developed in Wiesloch, Germany, is especially well suited to such patients. METHODS: Fifty-three mothers with postpartum disorders (33 depressive, 20 psychotic) were examined before and after therapy. Psychopathologic severity, psychosocial level of functioning, and parameters of the mother-child interaction were assessed and compared. RESULTS: Overall the results showed clear improvements in the assessed parameters at the end of treatment for both psychotic mothers and those with affective disorders. CONCLUSION: The interactional treatment program for postpartum mental disorders leads to a significant reduction in psychic/psychiatric severity and the associated psychosocial impairment and disturbed mother-child interaction. Considerations of the effects of therapy were not possible due to the study design. PMID- 17426952 TI - [Changes in the public's social distance toward individuals with schizophrenia in six German cities. Results of representative pre- and postinterventional telephone surveys from 2001 to 2004]. AB - BACKGROUND: In several German cities various programmes and initiatives have been conducted during recent years aimed at reducing the stigma attached to mental illness, especially schizophrenia. So far only single interventions in specific target groups have been evaluated but not their effectiveness in the general population. METHODS: Two representative telephone surveys with repeated measurements were conducted in six German cities (n 7,225/4,622). Social distance towards people with schizophrenia and knowledge of antistigma projects were assessed. RESULTS: Between 2001 and 2004 the social distance toward persons with schizophrenia decreased in the general population. Persons familiar with antistigma projects or who lived in cities with such projects showed lower social distance. CONCLUSIONS: For the first time evidence has been found that antistigma interventions are successful in reducing the stigma attached to schizophrenia in the general population. PMID- 17426953 TI - [Renal cell carcinoma--developments in therapy]. PMID- 17426954 TI - Mapping of five resistance genes to sugar-beet powdery mildew using AFLP and anchored SNP markers. AB - Sugar-beet powdery mildew, caused by the fungus Erysiphe betae, now occurs in all sugar-beet growing areas and can reduce sugar yield by up to 30%. Powdery mildew resistant plants from three novel sources were crossed with sugar beet to generate segregating populations. Evaluation of resistance was carried out in artificially inoculated field and controlled environment tests. The resistance level in two of the sources was found to be significantly higher than that in currently available sugar-beet cultivars. AFLP analysis was used in combination with bulked segregant analysis to develop markers linked to the resistant phenotype in each population. Five dominant major resistance genes were identified and assigned the proposed symbols Pm2 to Pm6. Pm3 conferred complete resistance to powdery mildew; the other genes conferred high levels of partial resistance. From the use of anchoring SNP markers, two genes were located to chromosome II and three to chromosome IV. Two of the genes on chromosome IV mapped to the same location and one of the genes on chromosome II mapped to the same region as the previously identified Pm1 gene. With the availability of these genes there is now excellent potential for achieving durable resistance to sugar beet powdery mildew, thus reducing or obviating the need for chemical control. PMID- 17426955 TI - Allelic variation of polyphenol oxidase (PPO) genes located on chromosomes 2A and 2D and development of functional markers for the PPO genes in common wheat. AB - Polyphenol oxidase (PPO) activity is highly related to the undesirable browning of wheat-based end products, especially Asian noodles. Characterization of PPO genes and the development of their functional markers are of great importance for marker-assisted selection in wheat breeding. In the present study, complete genomic DNA sequences of two PPO genes, one each located on chromosomes 2A and 2D and their allelic variants were characterized by means of in silico cloning and experimental validation. Sequences were aligned at both DNA and protein levels. Two haplotypes on chromosome 2D showed 95.2% sequence identity at the DNA level, indicating much more sequence diversity than those on chromosome 2A with 99.6% sequence identity. Both of the PPO genes on chromosomes 2A and 2D contain an open reading frame (ORF) of 1,731 bp, encoding a PPO precursor peptide of 577 amino acids with a predicted molecular mass of approximately 64 kD. Two complementary dominant STS markers, PPO16 and PPO29, were developed based on the PPO gene haplotypes located on chromosome 2D; they amplify a 713-bp fragment in cultivars with low PPO activity and a 490-bp fragment in those with high PPO activity, respectively. The two markers were mapped on chromosome 2DL using a doubled haploid population derived from the cross Zhongyou 9507/CA9632, and a set of nullisomic-tetrasomic lines and ditelosomic line 2DS of Chinese Spring. QTL analysis indicated that the PPO gene co-segregated with the two STS markers and was closely linked to SSR marker Xwmc41 on chromosome 2DL, explaining from 9.6 to 24.4% of the phenotypic variance for PPO activity across three environments. In order to simultaneously detect PPO loci on chromosomes 2A and 2D, a multiplexed marker combination PPO33/PPO16 was developed and yielded distinguishable DNA patterns in a number of cultivars. The STS marker PPO33 for the PPO gene on chromosome 2A was developed from the same gene sequences as PPO18 that we reported previously, and can amplify a 481-bp and a 290-bp fragment from cultivars with low and high PPO activity, respectively. A total of 217 Chinese wheat cultivars and advanced lines were used to validate the association between the polymorphic fragments and grain PPO activity. The results showed that the marker combination PPO33/PPO16 is efficient and reliable for evaluating PPO activity and can be used in wheat breeding programs aimed for noodle and other end product quality improvement. PMID- 17426956 TI - Over-expression of a LEA gene in rice improves drought resistance under the field conditions. AB - Late embryogenesis abundant (LEA) proteins have been implicated in many stress responses of plants. In this report, a LEA protein gene OsLEA3-1 was identified and over-expressed in rice to test the drought resistance of transgenic lines under the field conditions. OsLEA3-1 is induced by drought, salt and abscisic acid (ABA), but not by cold stress. The promoter of OsLEA3-1 isolated from the upland rice IRAT109 exhibits strong activity under drought- and salt-stress conditions. Three expression constructs consisting of the full-length cDNA driven by the drought-inducible promoter of OsLEA3-1 (OsLEA3-H), the CaMV 35S promoter (OsLEA3-S), and the rice Actin1 promoter (OsLEA3-A) were transformed into the drought-sensitive japonica rice Zhonghua 11. Drought resistance pre-screening of T(1) families at anthesis stage revealed that the over-expressing families with OsLEA3-S and OsLEA3-H constructs had significantly higher relative yield (yield under drought stress treatment/yield under normal growth conditions) than the wild type under drought stress conditions, although a yield penalty existed in T(1) families under normal growth conditions. Nine homozygous families, exhibiting over-expression of a single-copy of the transgene and relatively low yield penalty in the T(1) generation, were tested in the field for drought resistance in the T(2) and T(3) generations and in the PVC pipes for drought tolerance in the T(2) generation. Except for two families (transformed with OsLEA3-A), all the other families (transformed with OsLEA3-S and OsLEA3-H constructs) had higher grain yield than the wild type under drought stress in both the field and the PVC pipes conditions. No significant yield penalty was detected for these T(2 )and T(3) families. These results indicate that transgenic rice with significantly enhanced drought resistance and without yield penalty can be generated by over-expressing OsLEA3-1 gene with appropriate promoters and following a bipartite (stress and non-stress) in-field screening protocol. PMID- 17426957 TI - Consistent detection of QTLs for crown rust resistance in Italian ryegrass (Lolium multiflorum Lam.) across environments and phenotyping methods. AB - Crown rust, caused by Puccinia coronata f. sp. lolii, is one of the most important diseases of temperate forage grasses, such as ryegrasses (Lolium spp.), affecting yield and nutritional quality. Therefore, resistance to crown rust is a major goal in ryegrass breeding programmes. In a two-way pseudo-testcross population consisting of 306 Lolium multiflorum individuals, multisite field evaluations as well as alternative methods based on artificial inoculation with natural inoculate in controlled environments were used to identify QTLs controlling resistance to crown rust. Disease scores obtained from glasshouse and leaf segment test (LST) evaluations were highly correlated with scores from a multisite field assessment (r = 0.66 and 0.79, P < 0.01, respectively) and thus confirmed suitability of these methods for crown rust investigations. Moreover, QTL mapping based on a linkage map consisting of 368 amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers revealed similar results across different phenotyping methods. Two major QTLs were consistently detected on linkage group (LG) 1 and LG 2, explaining up to 56% of total phenotypic variance (V (p)). Nevertheless, differences between position and magnitude of QTLs were observed among individual field locations and suggested the existence of specific local pathogen populations. The present study not only compared QTL results among crown rust evaluation methods and environments, but also identified molecular markers closely linked to previously undescribed QTLs for crown rust resistance in Italian ryegrass with the potential to be applied in marker-assisted forage crop breeding. PMID- 17426958 TI - Joint modeling of additive and non-additive (genetic line) effects in multi environment trials. AB - A statistical approach for the analysis of multi-environment trials (METs) is presented, in which selection of best performing lines, best parents, and best combination of parents can be determined. The genetic effect of a line is partitioned into additive, dominance and residual non-additive effects. The dominance effects are estimated through the incorporation of the dominance relationship matrix, which is presented under varying levels of inbreeding. A computationally efficient way of fitting dominance effects is presented which partitions dominance effects into between family dominance and within family dominance line effects. The overall approach is applicable to inbred lines, hybrid lines and other general population structures where pedigree information is available. PMID- 17426959 TI - An ultradense genetic recombination map for Brassica napus, consisting of 13551 SRAP markers. AB - Sequence related amplified polymorphism (SRAP) was used to construct an ultradense genetic recombination map for a doubled haploid (DH) population in B. napus. A total of 1,634 primer combinations including 12 fluorescently labeled primers and 442 unlabeled ones produced 13,551 mapped SRAP markers. All these SRAPs were assembled in 1,055 bins that were placed onto 19 linkage groups. Ten of the nineteen linkage groups were assigned to the A genome and the remaining nine to the C genome on the basis of the differential SRAP PCR amplification in two DH lines of B. rapa and B. oleracea. Furthermore, all 19 linkage groups were assigned to their corresponding N1-N19 groups of B. napus by comparison with 55 SSR markers used to construct previous maps in this species. In total, 1,663 crossovers were detected, resulting in a map length span of 1604.8 cM. The marker density is 8.45 SRAPs per cM, and there could be more than one marker in 100 kb physical distance. There are four linkage groups in the A genome with more than 800 SRAP markers each, and three linkage groups in the C genome with more 1,000 SRAP markers each. Our studies suggest that a single SRAP map might be applicable to the three Brassica species, B. napus, B. oleracea and B. rapa. The use of this ultra high-density genetic recombination map in marker development and map-based gene cloning is discussed. PMID- 17426960 TI - C3H/HeJ mice carrying a toll-like receptor 4 mutation are protected against the development of insulin resistance in white adipose tissue in response to a high fat diet. AB - AIMS/HYPOTHESIS: Inflammation is associated with obesity and has been implicated in the development of diabetes and atherosclerosis. During gram-negative bacterial infection, lipopolysaccharide causes an inflammatory reaction via toll like receptor 4 (TLR4), which has an essential function in the induction of innate and adaptative immunity. Our aim was to determine what role TLR4 plays in the development of metabolic phenotypes during high-fat feeding. MATERIALS AND METHODS: We evaluated metabolic consequences of a high-fat diet in TLR4 mutant mice (C3H/HeJ) and their respective controls. RESULTS: TLR4 inactivation reduced food intake without significant modification of body weight, but with higher epididymal adipose tissue mass and adipocyte hypertrophy. It also attenuated the inflammatory response and increased glucose transport and the expression levels of adiponectin and lipogenic markers in white adipose tissue. In addition, TLR4 inactivation blunted insulin resistance induced by lipopolysaccharide in differentiated adipocytes. Increased feeding efficiency in TLR4 mutant mice was associated with lower mass and lower expression of uncoupling protein 1 gene in brown adipose tissue. Finally, TLR4 inactivation slowed the development of hepatic steatosis, reducing the liver triacylglycerol content and also expression levels of lipogenic and fibrosis markers. CONCLUSIONS/INTERPRETATION: TLR4 influences white adipose tissue inflammation and insulin sensitivity, as well as liver fat storage, and is important in the regulation of metabolic phenotype during a fat-enriched diet. PMID- 17426961 TI - A single high dose of cocaine induces differential sensitization to specific behaviors across adolescence. AB - RATIONALE: Adolescence is a critical period for drug addiction. Acute stimulant exposure elicits different behavioral responses in adolescent and adult rodents. The same biological differences that mediate age-specific behavioral responsiveness to stimulants in rodents could contribute to increased addiction vulnerability in adolescent humans. OBJECTIVES: This study compared the ability of a single high dose of cocaine (40 mg/kg) to induce behavioral sensitization to a challenge dose of cocaine (10 mg/kg) 24 h later in young adolescent postnatal day 28 (PN 28), mid-adolescent (PN 42), and young adult (PN 65) male rats. Horizontal activity was resolved into ambulatory and non-ambulatory movements. An observational behavioral rating was obtained by recording specific behaviors. We examined if individual behavioral responses to novelty and cocaine correlate with sensitization in each age group. RESULTS: Single dose cocaine pretreatment induced behavioral sensitization to non-ambulatory horizontal activity, sniffing behaviors, and stereotypies in animals of all ages. Ambulatory sensitization was observed only in the youngest adolescents. Cocaine pretreatment caused greater increases in stereotypies in the young adolescents than in adults. The magnitude of the behavioral response to the initial cocaine treatment was positively correlated with the magnitude of sensitization in individual young adolescents. High levels of novelty-induced ambulatory activity only correlated with the magnitude of ambulatory sensitization in the youngest adolescents. CONCLUSIONS: We conclude that a single high dose of cocaine produces age-specific patterns of behavioral sensitization. Young adolescent rats appear to be more sensitive than adults to some of the behavioral alterations induced by a single high dose of cocaine. PMID- 17426962 TI - Micromachined, planar-geometry, atmospheric-pressure, battery-operated microplasma devices (MPDs) on chips for analysis of microsamples of liquids, solids, or gases by optical-emission spectrometry. AB - Because of their desirable characteristics, for example small size, lightness, low power and gas consumption, and potential for portability, miniaturized plasma sources are receiving significant attention in the scientific literature. To take advantage of these characteristics we micromachined and fabricated new, planar geometry, self-igniting, atmospheric-pressure microplasma devices (MPDs) on chips. These microplasmas required such low power for their operation they could be operated from a re-chargeable battery (of the type used in cordless power tools). Despite their advantages, most miniaturized plasma sources reported in the literature have not performed well with liquid samples; analysis of powders or solids that can be converted to a powder (and processed and used as slurries) is even more difficult. To address these shortcomings we coupled an electrothermal, mini-in-torch vaporization (mini-ITV) "dry" sample-introduction system to the low-power planar microplasma devices we developed. In this preliminary investigation, absolute detection limits obtained from microsamples of single-element liquid standards and optical emission spectrometry with photomultiplier-tube detection and a spectral bandpass similar to that of portable, commercially available fiber-optic spectrometers were in the low-pg to ng range, for example 2 pg (for K) to 25 ng (for Pb). Mini-ITV also enabled (as far as we are aware, for the first time) measurement of analyte emission from microsamples of powdered solids (as slurries). In addition to the 3% H2 in Ar mixtures, the ac-operated microplasmas were sustained by use of a variety of electrode materials and different plasma-support gases (e.g. Ar, He and 3% H2 in He) thus indicating fabrication versatility and operational flexibility. Such flexibility has the potential to enable microplasmas to be tailored to analytical problems, and this is demonstrated by using a He MPD and chlorine emission measurements (837.594 nm) from gaseous microsamples as an example. PMID- 17426963 TI - Promotion from associate to full professor. PMID- 17426964 TI - Computer simulations of a dielectric barrier discharge used for analytical spectrometry. AB - A model developed for a dielectric barrier discharge (DBD) in helium, used as a new spectroscopic source in analytical chemistry, is presented in this paper. The model is based on the fluid approach and describes the behavior of electrons, He+ and He+2 ions, He metastable atoms, He atoms in higher excited levels, and He2 dimers. The He ground-state atoms are regarded as background gas. The characteristic effect of charging/discharging of the dielectrics which cover both electrodes is also simulated. Typical results of the model include the distribution of potential inside the plasma (and the potential drop across the dielectrics), the electric current and gap voltage as a function of time for a given applied potential profile, the spatial and temporal number-density profiles of the different plasma species, and the relative contributions of the mechanisms of their production and loss. PMID- 17426966 TI - Recombinant expression and characterization of XynD from Bacillus subtilis subsp. subtilis ATCC 6051: a GH 43 arabinoxylan arabinofuranohydrolase. AB - The complete genome sequence of Bacillus subtilis reveals that sequences encoding several hemicellulases are co-localised with a gene (xynD) encoding a putative family 43 glycoside hydrolase that has not yet been characterised. In this work, xynD has been isolated from genomic DNA of B. subtilis subsp. subtilis ATCC 6051 and cloned for cytoplasmatic expression in Escherichia coli. Recombinant XynD (rXynD) was purified using ion-exchange chromatography and gel permeation chromatography. The enzyme had a molecular mass of approximately 52 kDa, a pI above 9.0 and releases alpha-L-arabinose from arabinoxylo-oligosaccharides as well as arabinoxylan polymers with varying degree of substitution. Using para nitrophenyl-alpha-L-arabinofuranoside as substrate, maximum activity was observed at pH 5.6 and 45 degrees C. The enzyme retained its activity over a large pH range, while activity was lost after pre-incubation above 50 degrees C. Gas liquid chromatography and proton nuclear magnetic resonance spectrometry analysis indicated that rXynD specifically releases arabinofuranosyl groups from mono substituted C-(O)-2 and C-(O)-3 xylopyranosyl residues on the xylan backbone. As rXynD did not display endoxylanase, xylosidase or arabinanase activity and was inactive on arabinan, we conclude that this enzyme is best described as an arabinoxylan arabinofuranohydrolase. PMID- 17426967 TI - A Paenibacillus sp. dextranase mutant pool with improved thermostability and activity. AB - Random mutagenesis was used to create a library of chimeric dextranase (dex1) genes. A plate-screening protocol was developed with improved thermostability as a selection criterion. The mutant library was screened for active dextranase variants by observing clearing zones on dextran-blue agar plates at 50 degrees C after exposure to 68 degrees C for 2 h, a temperature regime at which wild-type activity was abolished. A number of potentially improved variants were identified by this strategy, five of which were further characterised. DNA sequencing revealed ten nucleotide substitutions, ranging from one to four per variant. Thermal inactivation studies showed reduced (2.9-fold) thermostability for one variant and similar thermostability for a second variant, but confirmed improved thermostability for three mutants with 2.3- (28.9 min) to 6.9-fold (86.6 min) increases in half-lives at 62 degrees C compared to that of the wild-type enzyme (12.6 min). Using a 10-min assay, apparent temperature optima of the variants were similar to that of the wild type (T (opt) 60 degrees C). However, one of these variants had increased enzyme activity. Therefore, the first-generation dextranase mutant pool obtained in this study has sufficient molecular diversity for further improvements in both thermostability and activity through recombination (gene shuffling). PMID- 17426968 TI - Flt3-L gene therapy enhances immunocytokine-mediated antitumor effects and induces long-term memory. AB - Therapeutic treatment with hu14.18-IL-2 immunocytokine (IC) or Flt3-L (FL) protein is initially effective at resolving established intradermal NXS2 neuroblastoma tumors in mice. However, many treated animals develop recurrent disease. We previously found that tumors recurring following natural killer (NK) mediated IC treatment show augmented MHC class I expression, while the tumors that recurred following T cell dependent Flt3-L treatment exhibited decreased MHC class I expression. We hypothesized that this divergent MHC modulation on recurrent tumors was due to therapy-specific immunoediting. We further postulated that combining IC and Flt3-L treatments might decrease the likelihood of recurrent disease by preventing MHC modulation as a mechanism for immune escape. We now report that combinatorial treatment of FL plus hu14.18-IL-2 IC provides greater antitumor benefit than treatment with either alone, suppressing development of recurrent disease. We administered FL by gene therapy using a clinically relevant approach: hydrodynamic limb vein (HLV) delivery of DNA for transgene expression by myofibers. Delivery of FL DNA by HLV injection in mice resulted in systemic expression of >10 ng/ml of FL in blood at day 3, and promoted up to a fourfold and tenfold increase in splenic NK and dendritic cells (DCs), respectively. Furthermore, the combination of FL gene therapy plus suboptimal IC treatment induced a greater expansion in the absolute number of splenic NK and DCs than achieved by individual component treatments. Mice that received combined FL gene therapy plus IC exhibited complete and durable resolution of established NXS2 tumors, and demonstrated protection from subsequent rechallenge with NXS2 tumor. PMID- 17426969 TI - Human NK cell infusions prolong survival of metastatic human neuroblastoma bearing NOD/scid mice. AB - AIM: Several lines of evidence suggest that NK cell immunotherapy may represent a successful approach in neuroblastoma (NB) patients refractory to conventional therapy. However, homing properties, safety and therapeutic efficacy of NK cell infusions need to be evaluated in a suitable preclinical murine NB model. MATERIALS AND METHODS: Here, the therapeutic efficacy of NK cell infusions in the presence or absence of NK-activating cytokines have been evaluated in a NB metastatic model set up in NOD/scid mice, that display reduced functional activity of endogenous NK cells. RESULTS: In NOD/scid mice the injected NB cells rapidly reached all the typical sites of metastatization, including bone marrow. Infusion of polyclonal IL2-activated NK cells was followed by dissemination of these cells into various tissues including those colonized by metastatic NB cells. The early repeated injection of IL2-activated NK cells in NB-bearing NOD/scid mice significantly increased the mean survival time, which was associated with a reduced bone marrow infiltration. The therapeutic effect was further enhanced by low doses of human recombinant IL2 or IL15. CONCLUSION: Our results indicate that NK-based adoptive immunotherapy can represent a valuable adjuvant in the treatment of properly selected NB patients presenting with metastatic disease, if performed in a minimal residual disease setting. PMID- 17426971 TI - Capecitabine and doxorubicin combination chemotherapy as salvage therapy in pretreated advanced gastric cancer. AB - PURPOSE: The aim of this study was to evaluate the activity and the safety of a combination regimen of capecitabine and doxorubicin as salvage chemotherapy in advanced gastric cancer patients who had undergone one or two prior chemotherapy regimens. METHODS: Patients received capecitabine, 2,500 mg/m(2)/day PO for 14 days (D1-14) and doxorubicin, 30 mg/m(2) IV on day 1 every 3 weeks until disease progression. The response was evaluated according to RECIST criteria, and the toxicity was evaluated by NCI-CTC (version 2.0). RESULTS: Forty-five patients were enrolled. Twenty-six patients were treated as second-line chemotherapy and the remaining patients as third-line chemotherapy. A total of 152 cycles of chemotherapy (median 2, range 1-12) were administered. Median dose intensities of capecitabine and doxorubicin were 11,326 and 9.6 mg/m(2)/week, respectively. The overall response rate was 6.7% (95% CI, 4.1-12.5%) and the disease control rate was 46.7% (95% CI, 28.6-87.1%) according to an intent-to-treat analysis. The median progression-free survival was 11.3 weeks (95% CI, 5.6-16.7 weeks). The median overall survival was 29.1 weeks (95% CI, 18.3-39.9 weeks) with one-year survival rate of 24%. Severe (grade III/IV) hematologic and non-hematologic toxicity was uncommon and included nausea/vomiting in five (11.1%), neutropenia in two (4.4%), anemia in one (2.2%), and hand-foot syndrome in one patient (2.2%). CONCLUSIONS: The combination of capecitabine and doxorubicin is a feasible salvage regimen in advanced pre-treated gastric cancer. PMID- 17426972 TI - Glutamine and alanyl-glutamine accelerate the recovery from 5-fluorouracil induced experimental oral mucositis in hamster. AB - INTRODUCTION: Mucositis induced by anti-neoplastic drugs is an important, dose limiting and costly side effect of cancer therapy. AIM: To evaluate the effect of oral glutamine and alanyl-glutamine, a more stable glutamine derivative, on 5-FU induced oral mucositis in hamsters. MATERIALS AND METHODS: Oral mucositis was induced by two intraperitoneal (i.p) administrations of 5-FU on the first and second days of the experiment (60 and 40 mg/kg, respectively) followed by mechanical trauma on the fourth day in male hamsters. Animals received saline, glutamine or alanyl-glutamine suspension (100 mM) 1 h before the injections of 5 FU and daily until sacrifice, on the 10th or 14th day. Macroscopic and histopathological analyses were evaluated and graded. Tissues from the cheek pouches were harvested for measurement of myeloperoxidase activity and glutathione stores. For investigation of serum concentration of glutamine, blood was obtained by heart puncture from anesthetized animals before sacrifice, on day 10. RESULTS: Treatment with glutamine and alanyl-glutamine reduced macroscopic and histological parameters of oral mucositis, and reduced the myeloperoxidase activity on day 14, but not on day 10. The 5-FU-induced oral mucositis significantly decreased the serum glutamine levels as well as the cheek pouch glutathione stores observed on day 10. Glutamine or alanyl-glutamine administration reversed the 5-FU effects, restoring serum glutamine levels and cheek pouch glutathione stores, observed on day 10, but did not prevent oral mucositis on the tenth day. CONCLUSION: Glutamine or alanyl-glutamine accelerated the mucosal recovery increasing mucosal tissue glutathione stores, reducing inflammatory parameters and speeding reepithelization. PMID- 17426973 TI - A phase I study of capecitabine and a modulatory dose of irinotecan in metastatic breast cancer. AB - PURPOSE: There is a need for chemotherapy regimens active against anthracycline- and taxane-refractory breast cancer. Data from preclinical and pilot studies performed at Roswell Park Cancer Institute (RPCI) suggested that when irinotecan (IRN) is given with 5-fluorouracil (5-FU) efficacy is affected by the sequence of drug administration. Pretreatment with IRN 24 h before 5-FU increased the number of tumor cells in S-phase and the antitumor activity in a preclinical system. These data provided the rationale for the evaluation of IRN and capecitabine, a 5 FU prodrug, sequentially administered in patients with metastatic breast cancer. The main objective of the study was to determine the MTD and identify dose limiting toxicities (DLTs) of capecitabine and IRN. Additionally, the degree of accumulation of cells in S-phase in tumor biopsies obtained at 24 h after the first dose of IRN was measured in consenting patients. PATIENTS AND METHODS: Metastatic breast cancer patients who experienced disease progression after at least one (taxane or anthracycline based) chemotherapy regimen and an expected survival of at least 3 months and ECOG performance status 0-2 were eligible. Twelve patients were enrolled and treated. The starting doses were IRN 80 mg/m(2) given over 90 min on days 1, 8, 22, 29, and capecitabine 1,500 mg/m(2)/day given days 2-15 and 23-36. Evaluation for response was performed after the first cycle. Sequential tumor biopsies were performed on five patients. RESULTS: The first three patients treated exhibited modulation in cyclin A index on tumor biopsy as defined by the study, defining the modulatory dose of IRN as 80 mg/m(2). Overall, 4/5 biopsies showed modulation. Dose Limiting Toxicities (DLTs) were assessed during the first cycle of therapy. Two DLTs (Grade 3 nausea vomiting and dehydration; grade 3 pneumonia, hypoxia, hypotension) were seen at dose level 2 of capecitabine (2,000 mg/m(2)/day) and the first cohort was expanded. There were no DLTs for patients treated at DL 1. No grade 3-4 toxicities occurred at DL 1. Seven patients were evaluable for response following one cycle of treatment (partial response 1, stable disease 4, progressive disease 2) Of the five inevaluable patients, two experienced DLT, one received 50% of the planned capecitabine dose, one progressed prior to evaluation, and one withdrew consent. CONCLUSION: IRN 80 mg/m(2) days 1, 8, 22, 29 in combination with capecitabine 1,500 mg/m(2)/day in divided dose days 2-15 and 23-36 has an acceptable toxicity profile and resulted in modulation of S-phase in 4/5 specimens examined. Further studies of the activity of this combination and modulatory effect of IRN are warranted. PMID- 17426974 TI - Sonodynamic therapy on chemically induced mammary tumor: pharmacokinetics, tissue distribution and sonodynamically induced antitumor effect of gallium-porphyrin complex ATX-70. AB - Sonodynamically induced antitumor effect of a gallium porphyrin complex, ATX-70 was evaluated on a chemically induced mammary tumor in Sprague-Dawley rats. The timing of 24 h after the administration of ATX-70 was chosen for ultrasonic exposure, based on pharmacokinetic analysis of ATX-70 concentrations in the tumor, plasma, skin, and muscle. At an ATX-70 dose not less than 2.5 mg/kg and at a free-field ultrasonic intensity not less than 3 W/cm(2), the synergistic effect between ATX-70 administration and ultrasonic exposure on the tumor growth inhibition was significant. These results suggest that ATX-70 is a potential sonosensitizer for sonodynamic treatment of spontaneous mammary tumors. PMID- 17426975 TI - Phylogeny and evolution of mating-type genes from Pyrenophora teres, the causal agent of barley "net blotch" disease. AB - The main aim of this study was to test the patterns of sequence divergence and haplotype structure at the MAT locus of Pyrenophora teres, the causal agent of barley 'net blotch' disease. P. teres is a heterothallic ascomycete that co occurs in two symptomatological forms, the net form (NF) and the spot form (SF). The mating-type genes MAT1-1-1 and MAT1-2-1 were sequenced from 22 NF isolates (12 MAT1-1-1 and 10 MAT1-2-1 sequences) and 17 SF isolates (10 MAT1-1-1 and seven MAT1-2-1 sequences) collected from Sardinian barley landrace populations and worldwide. On the basis of a parsimony network analysis, the two forms of P. teres are phylogenetically separated. More than 85% of the total nucleotide variation was found between formae speciales. The two forms do not share any polymorphisms. Six diagnostic nucleotide polymorphisms were found in the MAT1-1-1 intron (1) and in the MAT1-1-1 (3) and MAT1-2-1 (2) exons. Three diagnostic non synonymous mutations were found, one in MAT1-1-1 and two in MAT1-2-1. For comparison with P. teres sequence data, the mating-type genes from Pyrenophora graminea were also isolated and sequenced. Divergence between P. graminea and P. teres is of a similar magnitude to that between NF and SF of P. teres. The MAT genes of P. graminea were closer to those of SF than to NF, with the MAT1-2-1 SF peptide not different from the MAT1-2-1 peptide of P. graminea. Overall, these data suggest long genetic isolation between the two forms of P. teres and that hybridization is rare or absent under field conditions, with each form having some particular niche specialization. This indicates that research on resistance to P. teres should consider the two forms separately, as different species. PMID- 17426976 TI - Serum pro-matrix metalloproteinase-3 as an indicator of disease activity and severity in rheumatoid arthritis: comparison with traditional markers. AB - Matrix metalloproteinase-3 (MMP-3) production increases in rheumatoid arthritis (RA) and has been proposed as a marker of disease activity and joint damage. The aim of this cross-sectional study is to examine the usefulness of serum proMMP-3 as an indicator of disease activity and severity in comparison with erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP). Serum proMMP-3 was measured by a quantitative ELISA in 85 RA patients and 70 healthy subjects. Clinical and laboratory measures of disease activity and severity were obtained. Radiological joint damage was assessed by the method of Larsen. Serum proMMP-3 was significantly higher in RA patients than that in the healthy subjects. The active RA patients had significantly higher serum proMMP-3 than the inactive patients. Serum proMMP-3 was significantly correlated with some parameters of disease activity including swollen joints count, proximal interphalangeal joint score, morning stiffness, and Health Assessment Questionnaire; however, ESR and serum CRP were better correlated with all indicators of the disease activity than proMMP-3. The analysis of receiver operating characteristic supported that ESR and CRP had higher performance for reflection of activity compared to proMMP-3. There were no significant associations among Larsen score and proMMP-3, ESR, and CRP. Our results suggest that the cross-sectional measurement of serum proMMP-3 could not give additional information about RA disease activity compared to ESR and CRP, and could not give any information about joint damage. PMID- 17426977 TI - Late onset undifferentiated spondyloarthritis presenting with polymyalgia rheumatica features: description of seven cases. AB - To underline the importance of considering a diagnosis of undifferentiated spondyloarthritis (uSpA) in patients presenting polymyalgia rheumatica (PMR) features. All patients with late onset uSpA meeting criteria for PMR at the onset of their disease seen in the Rheumatology Division of Xeral-Calde Hospital of Lugo, Spain during a 5 year period, and in the Rheumatology Department of Lucania, Italy in a two and a half year period, were studied. Six patients with late onset uSpA showing PMR symptoms at the onset were seen during the study periods in the two centres. Another patient had previously been observed in Lugo in a study dealing with the spectrum of conditions mimicking PMR. Of the seven patients, five had manifestations of SpA at the beginning of the disease and two developed these in the following 6 months. All seven met the Amor and/or the ESSG criteria for classifying and diagnosing SpA. The possibility that late onset uSpA may have PMR-like features at the beginning of the disease should be taken into account. The diagnosis is not difficult if the entire clinical spectrum of SpA is considered. PMID- 17426978 TI - Metabolism of reactive oxygen species in cotton cytoplasmic male sterility and its restoration. AB - To elucidate reactive oxygen species (ROS) metabolism of cotton cytoplasmic male sterility and the effects of restorer gene on the metabolism of ROS, the metabolism changes in the production and scavenging of ROS and gene expression related to ROS-scavenging enzymes were investigated in the anther mitochondria of CMS line, maintainer line and hybrid F(1). During the abortion preliminary stage (sporogenous cell division stage), anthers of CMS line had a little higher superoxide (O(2)(-)) production rate and hydrogen peroxide (H(2)O(2)) and malondialdehyde (MDA) contents than those of maintainer or hybrid F(1. )Simultaneously, a little higher ROS contents might serve as a signal to increase the activity of superoxide dismutase (SOD) in anthers of CMS line to reduce the ROS damage to the anther development. But at the abortion peak (pollen mother cell meiosis stage), anthers of CMS line had extraordinarily higher ROS contents and lower ROS-scavenging enzymic activities compared with the hybrid F(1), during which the ROS contents and ROS-scavenging enzymic activities in hybrid F(1) were approximate to those of maintainer line. The expression of Mn-sod and apx mRNA in anther of CMS line was obviously inhibited when ROS produced with a great deal during anther abortion, however the gene expression in hybrid F(1) kept normal with the maintainer. Excessive accumulation of O(2)(*-) , H(2)O(2 )and MDA, significant reduction of ROS-scavenging enzymic activities and lower gene expression level of ROS-scavenging enzyme were coinstantaneous with male cells death in anthers of CMS line. But when the restorer gene was transferred into CMS line, excessive production of ROS could be eliminated in the anthers of hybrid F(1). The restorer gene likely plays an important role in keeping the dynamic balance between the production and elimination of ROS. PMID- 17426979 TI - Shoot formation from root tip region: a developmental alteration by WUS in transgenic tobacco. AB - We examined the effect of ectopic expression of WUS on the morphology of tobacco seedlings and the segments in vitro. WUS was amplified from Arabidopsis cDNA and introduced into the tobacco genome under the transcriptional control of the beta estradiol-inducible expression system. When 1-week-old transgenic seedlings were cultured in the presence of beta-estradiol, only the root tip region developed bulbous tissues followed by shoot formation and plant regeneration, suggesting its applicability for improving the strategy of micropropagation in recalcitrant species. Evident abnormality was not observed in the cotyledons, hypocotyl nor root except for the tip. However, ectopic WUS seemed to be functional in those parts through the observation of gene expression and the behavior of cultured segments. Small root segments with a root tip treated with beta-estradiol also showed bulbing but no shoots unless exogenous cytokinin was supplied. These findings suggest the existence of unknown factors regulating ectopic WUS function in the seedling. PMID- 17426980 TI - A pregnant lady with intermittent vaginal bleeding (2007: 3b). Placenta percreta. AB - Placenta percreta is a potentially life-threatening complication of pregnancy, which is increasing in incidence. Ante-natal diagnosis with ultrasound and magnetic resonance imaging aids the obstetric team in planning further management. We present a case of placenta percreta with imaging and a brief review of the literature. PMID- 17426981 TI - Improved body mass index after mesenterico-portal bypass. AB - Extrahepatic portal hypertension in children secondary to portal vein obstruction is frequently associated with impaired somatic growth. The aim of this study was to assess growth and nutritional status, as reflected by the body mass index (BMI), before and after mesenterico-portal bypass (Rex shunt). Eleven children with a portal vein cavernoma underwent mesenterico-portal bypass using autologous jugular vein. All shunts have remained patent during follow-up periods of 7 months to 5 years. All except one child, who had a normal BMI prior to surgery, demonstrated an increase in their BMI standard deviation scores after surgery. Mean BMI standard deviation scores increased from -0.44 +/- 1.28 (95% CI -1.30 to 0.42) to 0.46 +/- 1.08 (95% CI -0.27 to 1.19), a highly statistically significant increase (P=0.003). Restoration of hepatopetal portal blood flow by mesenterico portal bypass surgery improves nutrition and growth in children with extrahepatic portal hypertension. PMID- 17426983 TI - Introduction of the euro: the change is easily swallowed at a young age. PMID- 17426984 TI - Long-term cognitive and behavioral consequences of neonatal encephalopathy following perinatal asphyxia: a review. AB - Neonatal encephalopathy (NE) following perinatal asphyxia (PA) is considered an important cause of later neurodevelopmental impairment in infants born at term. This review discusses long-term consequences for general cognitive functioning, educational achievement, neuropsychological functioning and behavior. In all areas reviewed, the outcome of children with mild NE is consistently positive and the outcome of children with severe NE consistently negative. However, children with moderate NE form a more heterogeneous group with respect to outcome. On average, intelligence scores are below those of children with mild NE and age matched peers, but within the normal range. With respect to educational achievement, difficulties have been found in the domains reading, spelling and arithmetic/mathematics. So far, studies of neuropsychological functioning have yielded ambiguous results in children with moderate NE. A few studies suggest elevated rates of hyperactivity in children with moderate NE and autism in children with moderate and severe NE. CONCLUSION: Behavioral monitoring is required for all children with NE. In addition, systematic, detailed neuropsychological examination is needed especially for children with moderate NE. PMID- 17426985 TI - Healthcare technology and technology assessment. AB - New technology is one of the primary drivers for increased healthcare costs in the United States. Both physician and industry play important roles in the development, adoption, utilization and choice of new technologies. The Federal Drug Administration regulates new drugs and new medical devices, but healthcare technology assessment remains limited. Healthcare technology assessment originated in federal agencies; today it is decentralized with increasing private sector efforts. Innovation is left to free market forces, including direct to consumer marketing and consumer choice. But to be fair to the consumer, he/she must have free knowledge of all the risks and benefits of a new technology in order to make an informed choice. Physicians, institutions and industry need to work together by providing proven, safe, clinically effective and cost effective new technologies, which require valid pre-market clinical trials and post-market continued surveillance with national and international registries allowing full transparency of new products to the consumer--the patient. PMID- 17426986 TI - Reconstruction with fascia lata allograft of the posterior vertebra elements after resection for aneurysmal bone cyst in a child. AB - We report a case of ABC in a child where, after resection of the posterior spinal column of L1, we did a biological reconstruction using a posterior tension band with a segment of fascia lata allograft in tension between T12 and L2. After the long term follow up, X-ray and MRI controls showed a satisfactory alignment of the spine and no local recurrence. The patient now has no sign of spinal instability or deviation, with no kind of discomfort or pain, and has a normal life. In our experience this biological tension band interferes minimally with the growth of the spine, and has a less number of complications in comparison with other more aggressive methods and so is a good option for restoring the stability in young patients with benign spinal tumors that arises on the posterior column without having any kind of potential deviations. PMID- 17426987 TI - Treatment of cervical myelopathy in patients with the fibromyalgia syndrome: outcomes and implications. AB - Some patients with fibromyalgia also exhibit the neurological signs of cervical myelopathy. We sought to determine if treatment of cervical myelopathy in patients with fibromyalgia improves the symptoms of fibromyalgia and the patients' quality of life. A non-randomized, prospective, case control study comparing the outcome of surgical (n = 40) versus non-surgical (n = 31) treatment of cervical myelopathy in patients with fibromyalgia was conducted. Outcomes were compared using SF-36, screening test for somatization, HADS, MMPI-2 scale 1 (Hypochondriasis), and self reported severity of symptoms 1 year after treatment. There was no significant difference in initial clinical presentation or demographic characteristics between the patients treated by surgical decompression and those treated by non-surgical means. There was a striking and statistically significant improvement in all symptoms attributed to the fibromyalgia syndrome in the surgical patients but not in the non-surgical patients at 1 year following the treatment of cervical myelopathy (P or = 90 years) (3.1). Cartilage defects were found all over the joint surfaces, none of the five regions was more often affected than the others. Least osteophytes were found on the medial border of the facet joints. In conclusion, the prevalence of cervical facet joint degeneration is probably very high in individuals aged 50 years and more, with a tendency to increase in severity with age. All levels of the middle and lower cervical spine were affected to almost the same degree, whereas in the lumbar spine an increase in degeneration towards the lower levels was reported. Also, in the cervical spine in most cases the cartilage was evenly degenerated all over the joint surface while in the lumbar spine certain regions were reported to be affected predominantly. PMID- 17426993 TI - Chimpanzees really know what others can see in a competitive situation. AB - Chimpanzee's perspective-taking abilities are currently disputed. Here we show that in some food competition contexts, subordinate chimpanzees do take the visual perspective of dominant individuals, preferentially targeting a hidden piece of the food that the dominant cannot see over a piece that is visible to both individuals. However, the space where the animals compete is critical in determining whether subjects demonstrate this skill. We suggest that competition intensity, as mediated by these spatial factors, may play an important role in determining the strategy chimpanzees utilize in competitive contexts. Since some strategies may not require visual perspective taking in order to be successful, chimpanzees may not always demonstrate this skill. Differences in spatial arrangement may therefore account for the conflicting results of past studies. PMID- 17426994 TI - Development and characterization of biofilms on stainless steel and titanium in spent nuclear fuel pools. AB - The aim of the present research was to study the biofilms developed in a Spanish nuclear power plant and their ability to entrap radionuclides. In order to carry this out, a bioreactor, which was then submerged in a spent nuclear fuel pool, was designed. To characterise the biofilm on two different metallic materials (stainless steel and titanium), standard culture microbiological methods and molecular biology tools, as well as epifluorescence and scanning electron microscopy were used. The bacterial composition of the biofilm belongs to several phylogenetic groups (alpha, beta, and gamma-Proteobacteria, Actinobacteridae, and Firmicutes). The radioactivity of the biofilms was measured by gamma-ray spectrometry. Biofilms were able to retain radionuclides from radioactive water, especially (60)Co. The potential use of these biofilms in bioremediation of radioactive water is discussed. PMID- 17426995 TI - Plasma lipoproteins and triacylglycerol are predictors of small, dense LDL particles. AB - Recently published data suggest that the assessment of LDL subfraction profiles may contribute to the determination of the cardiovascular risk. In this study, we tested the ability of various metabolic parameters to estimate the presence or the preponderance of small, dense LDL particles (sdLDL). One hundred and fifty individuals attending the Outpatient Clinics of the University Hospital of Ioannina for suspected metabolic abnormalities were included in the study. Individuals were excluded if they were found to be diabetic or if they had a history of cardiovascular disease. Patients with thyroid dysfunction, liver or kidney diseases as well as those receiving drugs that may interfere with lipids or glucose metabolism were also excluded from the study. The ability of the various parameters to identify individuals with pattern B LDL phenotype or, alternatively, with measurable quantities of sdLDL particles was tested with the calculation of the areas under the ROC curves. The ratio of triglycerides to HDL C was the best predictor of the presence of the pattern B LDL phenotype. Nevertheless, when the variable of interest was the presence of measurable quantities of sdLDL subfractions, the ratio of apoB to apoAI had the best predictive ability. In conclusion the ratios of apoB to apoAI and of triglycerides to HDL-C can reliably predict the presence of measurable quantities of sdLDL particles and of the pattern B LDL phenotype, respectively. However, since the quantitative determination of sdLDL concentrations may contribute to the determination of the cardiovascular risk, whereas the role of the LDL particle size remains controversial, apoB to apoAI ratio could provide more valuable information compared to markers that simply predict the presence of the pattern B LDL phenotype. PMID- 17426996 TI - Apolipoprotein E epsilon4 genotype is independently associated with increased intima-media thickness in a recessive pattern. AB - Polymorphisms in the apolipoprotein E (Apo E) gene have been associated with lipid levels, carotid intima media thickness (CCA-IMT), inflammation and cardiovascular disease (CVD). Earlier findings suggested an association of the Apo E alleles with increased CCA-IMT following a recessive pattern. Whether associations might be independent of C-reactive protein (CRP), lipid levels and other CVD risk factors is not known. We investigated the relationships between Apo E (epsilon2, epsilon3 and epsilon4 alleles) and CCA-IMT, measured by B-mode ultrasound, in dominant and recessive models in a community-based sample of 437 men 75 years of age. In men homozygous for the epsilon4 allele CCA-IMT was significantly increased by 0.13 mm to 0.86 +/- 0.16 mm compared to 0.73 +/- 0.19 mm in non- epsilon4-carriers (P = 0.0012) and 0.73 +/- 0.21 mm in epsilon4 heterozygous (P = 0.0044) in unadjusted recessive models. The association between Apo E epsilon4 genotype and CCA-IMT was independent of Apo E epsilon2 and Apo E epsilon3 alleles, CRP, lipid variables (TG, LDL, HDL) and other CVD risk factors (smoking, hypertension, body mass index, diabetes) (P = 0.018). No relations between Apo E genotype and CCA-IMT were observed in dominant models. No significant associations between the Apo E epsilon2 and epsilon3 alleles and CCA IMT were found. In this study, men homozygous with the ApoE epsilon4 allele had thicker CCA-IMT, independently of Apo E epsilon2 and epsilon3 alleles, CRP, lipid variables (TG, LDL, HDL) and other CVD risk factors (smoking, hypertension, body mass index, diabetes), suggesting CCA-IMT to be modified by the ApoE epsilon4 genotype in a recessive pattern. PMID- 17426997 TI - Neurosurgical training and the spine: reflections on EANS winter meeting Luxembourg, February, 2006. PMID- 17426998 TI - Haemodynamic patterns in children with posttraumatic diffuse brain swelling. A preliminary study in 6 cases with neuroradiological features consistent with diffuse axonal injury. AB - BACKGROUND: In the present report we describe the cerebral haemodynamics and the neuroradiological findings observed in six consecutive children, three males and three females aged 4-15.6 yrs (mean age 8.95) displaying a neuroradiological pattern consistent with diffuse axonal injury (DAI) along with slit ventricles. METHODS: All the patients were admitted to the Paediatric Intensive Care Unit with GCS scores less than 8 after a severe brain injury. Serial head computed to mography (CT) and magnetic resonance (MR) scans demonstrated a radiological pattern of DAI. Transcranial Doppler Sonography (TCD) of the middle cerebral arteries was performed through the temporal bone window in all the patients. All patients but one underwent a continuous monitoring of intracranial pressure (ICP) and cerebral extraction of O(2) (CEO(2)). Treatment with barbiturates and hyperventilation was necessary in all the cases. In one patient, a bilateral decompressive cran iectomy was performed in order to decrease severe in tracranial hypertension. RESULTS: Hyperflow along with intracranial hyper tension, variably responsive to barbiturate medication, was observed in all the patients by means of TCD and CEO(2). CONCLUSIONS: Intracranial hypertension can be elevated in pediatric posttraumatic hyperflow syndromes associated with DAI. The observation of the time course of the parameters studied allowed us to modify the pharmacological treatment and/or perform surgical decompression (external cerebrospinal fluid (CSF) drainage in five cases; decompressive craniectomy in one case). Compartmental hyperflow TCD pattern was evident in only one patient. Although the limited number of pa tients in our series does not allow definite conclusions, we strongly believe that TCD, with ICP and CEO(2) monitoring, are useful tools in planning surgical strategy in children with neuroradiological signs of DAI. PMID- 17426999 TI - Adult cerebellar mutism and cognitive-affective syndrome caused by hemangioblastoma. PMID- 17427002 TI - [Vaccination--the big success story]. PMID- 17427003 TI - [History of recommendations for immunizations in Austria]. AB - Public immunizations against small-pox were first conducted in Austria over 200 years ago on December 10th 1800. The first legal regulation concerning small-pox vaccination was issued in 1836. Official recommendations for childhood vaccinations were first published in 1973 in conjunction with the Austrian anticipatory health care program for children. Since 1984 the Ministry of Health in cooperation with the Federal Advisory Committee on Health publishes and updates recommendations for immunizations for all age groups at intervals dictated by epidemic necessities and availability of vaccines. PMID- 17427004 TI - Influenza vaccination in Austria, 1982-2003. AB - Since the past decade influenza vaccination is becoming an increasingly important aspect of public health programs. In the early 1990s independent investigators began to gather information on the comparative use of influenza vaccine in developed countries. The annual level of influenza vaccine distributed in each country was calculated as the number of doses distributed per 1000 total resident population. During the first period of research in 18 developed countries influenza vaccine was widely underused. From 1980 to 1992 the annual vaccination rates increased in most of the 18 countries. But Austria (Switzerland and Finland) showed little change (20 doses/1000 in 1982 and 23 doses in 1992). When in 1992-1995 four new countries were included, doses increased slightly in Austria, too, to about 52 doses/1000 in 1995, but the country still belonged to the low-use countries compared to Spain 160 doses/1000, although vaccination was recommended for all elderly people <65 years and persons with high-risk medical conditions. Self-payment was usual, and there was no reimbursement within the national or social health insurance schemes. From 1996 (77 doses/1000), to 2000 (118 doses/1000) and 2003 (127 doses/1000), vaccination increased slightly, but Austria remained one of the lowest three Western European countries reported, together with Sweden (127) and Norway (102). Possible reasons for the low usage in Austria may be the following: people mistake influenza for an influenza-like illness, a lack of effective social marketing, costs are not taken over by social or private insurance and discordance in the Austrian medical fraternity about the importance of vaccination. Especially in view of a possible new influenza pandemic, public awareness of the importance of vaccination must increase. PMID- 17427005 TI - [Examples for vaccines against diarrheal diseases--rotavirus and traveller's diarrhea]. AB - Diarrheal diseases constitute one of the most important health problems worldwide. Children less than 5 years, living in developing countries, are particularly in danger with respect to the incidence and severity of the gastrointestinal disorders. Travelers to developing countries are also at risk to develop diarrheal disorders; around 30-50% of them acquire so called "travelers's diarrhea" caused by bacteria, viruses or protozoa. It has been estimated that approximately 30-70% of diarrhea are due to bacteria, of which the most frequently detected enteric pathogens are non-invasive, enterotoxigenic Escherichia coli (ETEC). Their exotoxins, the heat stabile (ST) and the heat labile (LT) toxins are in large part responsible for the pathogenicity of the bacteria. About 20% of cases of traveler's diarrhea are caused by LT producing ETEC. This heat labile toxin exhibits a 80% sequence homology with cholera toxin. The presently available vaccine against cholera (Dukoral) contains inactivated Vibrio cholerae bacteria and the recombinant non-toxic B subunit of cholera toxin. Consequently, this vaccine displays also some efficacy against traveler's diarrhoea with up to 25 % of travelers being protected against this disease. Rotaviruses are the leading recognized cause of diarrhoea-related illness and deaths among infants worldwide in developing and industrialized countries. Based on the high incidence of this disease two oral vaccines have been developed and are available in Europe in 2007. Due to the impact of rotavirus diseases also in Austria vaccination against this disease has been already suggested in the Austrian vaccination schedules for infants from 6-24 weeks of age. One of the two vaccines, Rotarix, is an attenuated monovalent vaccine with a broad cross reactivity against the most frequent serotypes. The second one, RotaTeq, is a pentavalent attenuated vaccine containing 5 human-bovine reassortants. Both vaccines display 85-98% efficacy against severe rotavirus disease and an excellent tolerability with no difference in side reactions to the placebo controls, particularly with respect to intussusceptions. PMID- 17427006 TI - [FSME monitoring: monitoring of adverse events of tick-borne-encephalitis vaccines by selected paediatricians and general practitioners]. AB - The Institute for Vaccine-Safety of the Austrian Green Cross, the so-called "Institut fur Impfsicherheit des Osterreichischen Grunen Kreuzes", promoted a postmarketing sentinel study referring to the safety of tick-borne-encephalitis vaccines in spring 2002. Five-hundred selected paediatricians and general practitioners took part and generated data on 25,905 administered doses. They reported on 107/25,907 (0.413 %) local and systemic adverse reactions. The leading side-effect was fever, besides other adverse reactions corresponding to those known in the literature. The age-group most often affected was that of toddlers and infants. Monitoring with the help of selected paediatricians and general practitioners, carried out alongside the routine obligatory procedures of monitoring, proved to be a valid instrument for detecting vaccine adverse events of tick-borne-encephalitis vaccines. PMID- 17427007 TI - [Genetic immunization: new ways for protective and therapeutic vaccines against allergic diseases]. AB - Gene vaccines have proven to be a powerful tool to induce anti-allergic immune responses. Their underlying functional principle is based on the recruitment of allergen-specific Th1 cells and the establishment of a Th1 cytokine milieu, which protects against the development of a Th2-biased response and balances an already ongoing Th2-type response. Genetic immunization also offers novel approaches to the major problems associated with protein immunization, such as crosslinking of preexisting IgE on mast cells/basophils or induction of de novo synthesis of IgE by the protein immunization itself. In addition to the description of the principles of gene vaccination, this review gives a short overview of recently developed anti-allergic gene vaccines with an optimized efficacy and safety profile. PMID- 17427008 TI - Immunodominant peptides from conserved influenza proteins--a tool for more efficient vaccination in the elderly? AB - Influenza-specific CD8+ T cells are important for the clearance of infection especially in high risk groups such as elderly persons. Activation of these cells by immunization might therefore be a useful tool for a better protection of this specific age group. We therefore analyzed the frequency, phenotype and function of CD8+ T cells with specificity to the influenza M1(58-66) peptide in young, middle-aged and elderly persons ex vivo and after in vitro stimulation. Significantly lower numbers of M1(58-66)-specific CD8+ T cells were detected in the middle-aged and elderly compared to young donors. M1(58-66)-specific CD8+ T cells were either CD45RA(low)CD45RO(low) or CD45RA-CD45RO+, expressed CD28 and CD62L and did not produce perforin. There was no difference in the phenotype of influenza-specific CD8+ T cells between the three age groups. Despite the initially low numbers of M1(58-66)-specific CD8+ T cells in the older age groups, these cells could be expanded in vitro following peptide stimulation. They also acquired a CD45RO+CD28+ CD62L(+/-) phenotype and produced perforin. Our results demonstrate that although initially low in number, M1(58-66)-specific CD8+ T cells from elderly persons can be propagated and differentiated into perforin producing effector cells upon appropriate stimulation. M1(58-66) peptide or other immunodominant peptides derived from conserved influenza proteins could therefore be useful in future influenza vaccines in order to render elderly persons better protected against disease, in particular in the case of an influenza pandemic. PMID- 17427009 TI - Insights into molecular medicine: development of new diagnostic and prognostic parameters. AB - Molecular medicine leads us towards an understanding of some diseases at the molecular level. Examples are the analysis of immune complexes and receptor antireceptor compounds used in clinical medicine. Structural changes of some serum proteins occur in inflammation, neoplasia and autoimmunity. The detection and analysis of such structural modifications may offer a new field for the diagnosis, prognosis and therapy of some diseases. Modern medicine requires new technologies with high sensitivity, specificity and applicability. For the first time in Austria we have combined fluorescence correlation spectroscope (FCS), surface enhanced laser desorption ionisation--time of flight (SELDI-TOF) and the molecular modelling and visualization system according to the computer enhanced programs. Experimental and computational methods are combined in such a way that clinical data can be interpreted by theoretical methods at a molecular level or vice versa, the computational output delivers input for new investigations. One method brings us single results. In view of the spectrum of parameters relevant to clinical entities, multiplexing is a new way of development. Since the technologies are new, the scientifically interested reader should be informed about the matters arising. PMID- 17427010 TI - [Cervix carcinoma--Austrian Vaccination Committee recommends HPV vaccination]. PMID- 17427011 TI - [2006 immunization plan--details of rotavirus vaccination]. PMID- 17427014 TI - [Food borne infections: study of outbreaks--the key to the source]. PMID- 17427015 TI - [Informed consent in medical treatment of minors and revised regulation of guardianship]. PMID- 17427016 TI - Giant lipoma of the thenar. PMID- 17427017 TI - [Food borne infectious outbreaks, Austria 2005]. AB - In Austria, a total of 606 food borne outbreaks, affecting 1,910 people altogether (including 368 hospitalized patients and 1 fatal outcome) were documented in 2005. Salmonella spp. and Campylobacter spp. accounted for 99% of all reported outbreaks. Fortysix (8%) of the 606 outbreaks were acquired abroad. Bacteria caused all but one of the 560 domestically acquired food borne outbreaks: 427 (76%) were due to Salmonella spp., 128 (23%) due to Campylobacter spp. and two outbreaks each due to enterohaemorrhagic Escherichia coli and Yersinia enterocolitica. Norovirus caused an outbreak affecting 22 persons. The respective hospitalization rate for domestically acquired Salmonella spp. and Campylobacter spp. infections was 20% and 16%. Of those outbreaks where information as to the source was provided, eggs were implicated in 57%, meat products (especially poultry) in 30% and milk or dairy products (especially raw milk) in 4%. The ratio between family outbreaks and general outbreaks was 89% to 11%. For general outbreaks the following locations of exposure were given: commercial food suppliers (incl. restaurants, cafeterias) 34 x, family celebrations 14 x, nursery schools 4 x, festivities (e.g. fairs) 3 x, nursing homes twice and once a mixed outbreak involving commercial food suppliers plus homes (Austria-wide S. Enteritidis PT19 outbreak). In our opinion, the relatively high number of family outbreaks merely reflects the still insufficient quality of epidemiological outbreak investigation in Austria, i.e. lack of consolidating individual clusters into larger food borne outbreaks which exceed district or provincial borders. PMID- 17427018 TI - Economic growth and the incidence of occupational injuries in Austria. AB - OBJECTIVES: The aim of this paper was to analyze the impact of economic growth measured by real gross domestic product (GDP) on the incidence of occupational injuries in Austria. METHODS: The relationship between GDP and the occupational injury rate of the wage-earning population between 1955 and 2004 was analyzed using an error correction model. The sample size increased from 1.568,371 persons in 1955 to 2.656,952 in 2004. Occupational injuries were divided into fatal and non-fatal injuries. RESULTS: Occupational injuries (fatal and non-fatal) decreased from 8.59% to 4.08%: non-fatal injuries decreased from 8.56% to 4.07%; fatal injuries decreased from 0.03% to 0.01%. Austrian GDP increased from EUR 37.7 billion to EUR 202.8 billion (base year 1995). Statistical analysis clearly shows that a growing economy is associated with declining injury rates (fatal and non-fatal). Two mechanisms are discussed. Firstly, rising GDP is accompanied by greater investment in safer technologies and occupational safety measures. Secondly, booming economies are associated with a reduced risk of unemployment, which is already known to be a risk factor for occupational injuries. CONCLUSIONS: Economic development appears to have an impact on the incidence of occupational injuries in Austria. Health policy should emphasize the necessity for safety at work particularly in phases of economic slowdown. PMID- 17427019 TI - Family medicine practice and research: survey of physicians' attitudes towards scientific research in a post-communist transition country. AB - BACKGROUND: Although the paradigm of modern medicine is evidence-based practice, there is a lack of research output and interest in research in family medicine. We investigated attitudes towards scientific research among family medicine practitioners in a country in post-communist socioeconomic transition, and related it to their attitudes towards alternative medicine and clinical knowledge relevant for their practice. METHODS: We surveyed 427 family medicine practitioners in Croatia about their attitudes towards scientific research (5 point rating scale, total score range 20-100) and alternative medicine (5-point rating scale, total score range 14-70). We also tested their knowledge on diagnosis and treatment of hypertension (10 questions) and diabetes (12 questions). RESULTS: The attitude towards scientific research was positive (score 79.0 +/- 7.2 out of maximum 100) and significantly more positive than that towards alternative medicine (score 45.0 +/- 9.9 out of maximum 70; t(425) = 19.06, P < 0.001). The respondents correctly answered about half the questions on hypertension and diabetes; knowledge on new diagnostic and treatment guidelines was better than their textbook knowledge. The attitude scores were not related to knowledge or research activity or the medical practice of the respondents. CONCLUSIONS: Family medicine practitioners in a transition country have a more positive attitude towards science than towards alternative medicine, despite the adverse situation in which they practice. To involve family medicine practitioners in research, interventions must be directed towards changes in behavior and practice and not only towards increasing positive attitudes. PMID- 17427020 TI - Impaired anal sphincter function in professional cyclists. AB - BACKGROUND: The purpose of our study was to assess anal function in professional mountain bikers, as an increasing number of cycling enthusiasts report functional and morphological problems in the anal region. We tried to find out if constant saddle vibration suffered by professional cyclists could lead to defecation disorders. METHODS: 19 professional male mountain bikers took part in this study. The findings were compared with those of a control group (CG) of healthy volunteers who rode bikes less frequently. Both groups underwent rectal examination and anorectal manometry. Rectal sensation, rectal compliance and the anorectal inhibitory reflex were also assessed. Information on anal pain, blood on the toilet tissue, history of hemorrhoids, anal fissure, abscess formation and problems with defecation was collected by questionnaire and local examination was carried out. RESULTS: Sphincter resting pressures (89 +/- 27 mmHg vs CG: 69 +/- 16 mmHg), squeeze pressures (137 +/- 36 mmHg vs CG: 96 +/- 19 mmHg) and sphincter volumes (resting vector volume: 513 +/- 277 cc vs CG 273 +/- 170 cc, squeeze vector volume: 1389 +/- 867 cc vs CG: 490 +/- 219 cc) were significantly higher in the professional mountain bikers. Values reflecting rectal sensation (sensory threshold: 51 +/- 32 ml vs CG: 46 +/- 12 ml) were not significantly different between the groups. CONCLUSION: Proctological problems are very common in professional mountain bikers. Permanent microtrauma through constant saddle vibration may lead to chronic inflammation and anal fissure resulting in anal pain and therefore high sphincter pressure. Muscle hypertrophy as a consequence may lead to defecation disorders. PMID- 17427021 TI - Suicidal ideation among Viennese high school students. AB - OBJECTIVE: Suicidality spans a spectrum ranging from suicidal thoughts to suicidal acts, and suicidal ideation is more prevalent in youth populations, suggesting important developmental issues. We assessed the prevalence of lifetime suicidal ideation and associated psychosocial factors among Viennese high school students. METHOD: An anonymous self-report survey assessing demographic characteristics and the major psychiatric risk factors of teenage suicide was completed by students at three Viennese high schools (n = 214; mean age 15.4 years). RESULTS: Eighty-one (37.9%) high school students reported having had suicidal thoughts at some point in their lives, girls significantly more often than boys (48.5% vs. 29.1%, P = 0.004). Furthermore, lifetime suicidal ideation in Viennese high school students was associated with living in broken-home families, cigarette smoking, substance problems (alcohol/drugs), self-reported depression, and high school type (the highest prevalence was in grammar school). After adjusting for confounders, we found that female gender, substance problems, school type and cigarette smoking were significantly associated with lifetime suicidal ideation in Viennese high school students. CONCLUSIONS: The psychosocial factors associated with adolescent suicidal ideation require attention in the contexts of suicide prevention and mental health promotion. PMID- 17427022 TI - Subtype and genotypic resistance analysis of HIV-1 infected patients in Austria. AB - BACKGROUND: Analysis of HIV-1 subtypes and genotypic resistance have been shown to be relevant for epidemiologic and therapeutic studies or for vaccine development. In Europe, the majority of HIV-1 isolates belong to subtype B. Due to migration an increasing incidence for additional subtypes and complex recombinant forms are expected. OBJECTIVES AND STUDY DESIGN: To evaluate the prevalence of HIV-1 subtypes in Austria, 188 plasma samples of treatment experienced patients were investigated. For phylogenetic analysis protease and reverse transcriptase genes were amplified and sequenced. Subtypes were determined by comparing reference sequences. For genotypic resistance determination, the Resistance-Algorithm-Comparison from Stanford University was used. RESULTS: Non-B subtypes were found in 20.2% of all patients with a dominant prevalence (50%) in the Southern provinces of Austria. With 85% CRF01_AE and CRF02_AG are the predominant circulating recombinant forms in Austria. When resistance mutations were analyzed, 57.4% of all patients were susceptible to all three groups of antiretroviral drugs, whereas in 12.2% resistance against all three classes of antiretroviral drugs was found. CONCLUSION: HIV-1 subtype B is still dominant in major parts of Austria. However, a significantly increasing percentage of non-B subtypes and recombinant forms are observed in the Southern provinces. PMID- 17427023 TI - Adventitial cystic disease of the popliteal artery: a potential cause of intermittent claudication. AB - Adventitial cystic disease (ACD) is an uncommon and rare vascular entity characterized by a highly viscous and mucinous cyst located in the adventitia of an artery. Although first described in the external iliac artery, the disease is more frequently sited in the popliteal artery. ACD characteristically occurs in young adults, mostly male, with intermittent claudication as the initial presenting symptom. As the cyst enlarges, it leads to vascular compression with stenosis or occlusion. Because cystic formation of the popliteal artery is uncommon, the disease is often unrecognized. We report a case of popliteal artery ACD in a 54-year-old woman who was successfully treated surgically. PMID- 17427024 TI - [Evidence and consensus based Austrian guidelines for management of acute and chronic nonspecific backache]. PMID- 17427025 TI - Cerebrospinal fluid leaks. PMID- 17427027 TI - Gap junction coding genes and schizophrenia: a genetic association study. AB - The aim of this study was to evaluate the association of genes that encode gap junction forming proteins and schizophrenia. Representative genetic candidates (Panx2 and Cx36) from two families of gap junction genes were selected for analysis. According to the present findings these genes represent both functional and positional candidates for schizophrenia. The sample was comprised of 381 schizophrenic patients, and the same number of matched controls was tested in this study in order to evaluate the possible influence of the aforementioned genes on the pathogenesis of schizophrenia. Four SNPs in the case of Panx2 and two SNPs in the case of Cx36 were selected for analysis. Allele-, genotype- and haplotype-wise association did not yield statistically significant results. These data do not suggest that Panx2 or Cx36 could increase the risk of schizophrenia in the Japanese population. PMID- 17427028 TI - Prediction of complex traits based on the epistasis of multiple haplotypes. AB - Analysis of epistasis, or gene-gene interactions, is of particular importance for revealing the molecular mechanisms of complex human diseases. Multiple genes, each of which has a moderate effect, might interact and produce a complex phenotypic trait. In this paper, we present a novel method of epistasis analysis, utilizing multiple phase-resolved haplotypes residing in different genomic regions. Prediction models can then be derived from the epistasis to indicate the susceptibility of a person to a dichrotomous phenotypic trait. The simulation results showed that the prediction accuracy of this method is dependent on the penetrance rate of the underlying model. The computation cost, on the other hand, is dependent on the number of genomic regions involved for the complex phenotypic trait. PMID- 17427029 TI - A novel DFNA5 mutation does not cause hearing loss in an Iranian family. AB - Mutations in DFNA5 lead to autosomal dominant non-syndromic sensorineural hearing loss that starts at the high frequencies. To date, only three DFNA5 mutations have been described, and although different at the genomic DNA level, all lead to exon 8 skipping at the mRNA level. This remarkable fact has led towards the hypothesis that DFNA5-associated hearing loss is caused by a gain-of-function mutation and not by haplo-insufficiency as previously thought. Here, we describe a fourth DFNA5 mutation: the insertion of a cytosine at nucleotide position 640 (AF073308.1:_c.640insC, AAC69324.1:_p. Thr215HisfsX8). Unlike the previously described mutations, this frameshift mutation truncates the protein in exon 5 of the gene. Although the mutation was found in an extended Iranian family with hereditary hearing loss, it does not segregate with the hearing loss phenotype and is even present in persons with normal hearing. This fact provides further support for the hypothesis that DFNA5-associated hearing loss is caused by a gain of-function mutation. PMID- 17427030 TI - Mutation identification and characterization of a Taiwanese patient with fucosidosis. AB - Fucosidosis is a rare lysosomal storage disease caused by a defect of the alpha L: -fucosidase (FUCA1) gene. Worldwide 26 mutations underlying the disease have been reported. By direct DNA sequencing of exons and flanking introns, homozygous Y126X mutation and Q281R polymorphism were found in a Taiwanese patient with fucosidosis. Upon expressing in COS-7 cells, 97.4% of alpha-L: -fucosidase activity compared with that of the wild-type construct was observed in the cDNA containing Q281R polymorphism. Western blot analysis revealed a 58-kDa precursor and 56-kDa mature forms for cells transfected with wild-type and Q281R enzymes. Using the fluorogenic substrate, the Michaelis constants and maximal velocities of both enzymes were very similar. While no appreciable enzyme activity (0.0%) was observed with Y126X mutation, no apparent decrease in FUCA1 mRNA level was seen with Y126X mutation. The expressed truncated Y126X protein was unstable and largely degraded. The delineation of the molecular defect could serve to complement future prenatal diagnosis for this family when necessary. PMID- 17427031 TI - Mutation analysis and genotype/phenotype relationships of Gaucher disease patients in Spain. AB - Mutations in the glucocerebrosidase (GBA) gene cause Gaucher disease (GD). The aim of this study was to characterise the GBA mutations and analyze genotype/phenotype relationships in 193 unrelated patients from the Spanish GD Registry. We have identified 98.7% of the mutated GBA alleles, finding 56 different GBA mutations and 66 genotypes causing GD in Spain: 47 previously described mutations and 9 novel mutations (4 missense R395C, R463H, W312R and V398I, 1 nonsense R359X, 4 frameshift c.708delC, c.1214-1215delGC, c.1439 1445del7 and c.42-65del24). The most prevalent mutations were N370S and L444P, accounting for 68.7% of the mutated alleles. A wide phenotypic difference was observed within each genotypic group, and 9% of diagnosed type 1 patients developed neurological involvement including parkisonism, tremor, hypoacusia and eye movements. All of these findings indicate that there is a significant genotypic heterogeneity that explains the huge phenotypic variation among Spanish GD patients. PMID- 17427032 TI - Cytochrome P450 1A1 (CYP1A1) T3801C and A2455G polymorphisms in breast cancer risk: a meta-analysis. AB - The cytochrome P450 1A1 gene (CYP1A1), encoding Phase I metabolic enzymes, appeared to be a candidate gene for breast cancer risk. However, studies on the association between polymorphisms in this gene and breast cancer have yielded conflicting results. We performed a meta-analysis to investigate the association with breast cancer of the CYP1A1 polymorphisms T3801C (9,316 cases and 12,714 controls) and A2455G (9,552 cases and 9,320 controls). In the genotype contrast of A2455G, both additive [GG vs AA, P = 0.04, fixed-effects OR 0.72; 95% CI (0.53 0.99), P = 0.95 for heterogeneity] and recessive [GG vs (GA + AA), P = 0.04, fixed-effects OR 0.73; 95% CI (0.53-0.99), P = 0.97 for heterogeneity] models produced significant results in east-Asians. In pre-menopausal women in a worldwide population, significant association between A2455G and breast cancer was also found using both models [additive model: P = 0.02, fixed-effects OR 0.52; 95% CI (0.29-0.92), P = 0.39 for heterogeneity; recessive model: P = 0.02, fixed-effects OR 0.51; 95% CI (0.29-0.90), P = 0.38 for heterogeneity]. Our meta analysis suggests that an A2455G G/G genotype is associated with a trend of reduced breast cancer risk, both in east-Asian women and in pre-menopausal women worldwide, while the T3801C C allele might not be a risk factor for breast cancer. Larger scale primary studies are required to further evaluate the interaction of CYP1A1 polymorphisms and breast cancer risk in specific populations. PMID- 17427033 TI - Development and structure of the female gametophyte in Austrobaileya scandens (Austrobaileyaceae). AB - Austrobaileyales, comprising the four families Austrobaileyaceae, Trimeniaceae, Schisandraceae, and Illiciaceae, are included in the basal angiosperms along with Amborellaceae and Nymphaeaceae. Here, we present the first developmental study of the female gametophyte in Austrobaileya scandens, the only species of Austrobaileyaceae, which are sister to the rest of the Austrobaileyales. Austrobaileya scandens has a four-celled/four-nucleate embryo sac as in the derived families of the order, e.g., Illiciaceae and Schisandraceae. It is monosporic, with the chalazal megaspore of a tetrad developing into the embryo sac composed of an egg cell, two synergids, and one polar nucleus. This mode of embryo sac formation was first reported in Schisandra over 40 years ago and should now be established as the Schisandra type. Its occurrence in A. scandens shows that the Schisandra-type embryo sac is likely common to the whole Austrobaileyales as well as to Nymphaeaceae. Amborellaceae were recently reported to have an eight-celled/nine-nucleate embryo sac, clarifying that none of the basal angiosperms has the seven-celled/eight-nucleate Polygonum-type embryo sac found in the majority of angiosperms, and that the Polygonum-type embryo sac represents a derived character state in angiosperms. PMID- 17427034 TI - Disruption of the psbA gene by the copy correction mechanism reveals that the expression of plastid-encoded genes is regulated by photosynthesis activity. AB - The functional analysis of genes encoded by the chloroplast genome of tobacco by reverse genetics is routine. Nevertheless, for a small number of genes their deletion generates heteroplasmic genotypes, complicating their analysis. There is thus the need for additional strategies to develop deletion mutants for these genes. We have developed a homologous copy correction-based strategy for deleting/mutating genes encoded on the chloroplast genome. This system was used to produce psbA knockouts. The resulting plants are homoplasmic and lack photosystem II (PSII) activity. Further, the deletion mutants exhibit a distinct phenotype; young leaves are green, whereas older leaves are bleached, irrespective of light conditions. This suggests that senescence is promoted by the absence of psbA. Analysis of the transcript levels indicates that NEP (nuclear-encoded plastid RNA polymerase)-dependent plastid genes are up regulated in the psbA deletion mutants, whereas the bleached leaves retain plastid-encoded plastid RNA polymerase activity. Hence, the expression of NEP-dependent plastid genes may be regulated by photosynthesis, either directly or indirectly. PMID- 17427035 TI - A mixture model approach to the mapping of QTL controlling endosperm traits with bulked samples. AB - Endosperm traits are of triploid inheritance and have become a focus of breeding effort for their close relations with the grain quality. Current methods for mapping quantitative trait loci (QTL) underlying endosperm traits are restricted to the use of the phenotypes of single grain samples as input data set, which are often not available in practice due to the small size of the cereal seeds. This paper proposed a statistical model for one specially tailored mapping strategy, where the marker genotypes are obtained from the maternal plants in the segregation population and the phenotypic responses are replaced by the trait means of composite endosperm samples pooled from each plant. It should therefore be more practical and have wide applicability in mapping endosperm traits. The method was implemented by fitting the phenotypic means of endosperms into a Gaussian mixture model. Both the exact and approximate Expectation-Maximization algorithms were proposed to estimate the model parameters. The presence of the QTL was determined by likelihood ratio test statistics. Statistical power and other properties of the new method were investigated and compared to the current single-seed method under a variety of scenarios through simulation studies. The simulations suggest a reasonable sample size should be used to ensure reliable results. The proposed method was also applied to a simulated genome data for further evaluation. As an illustration, a real data of maize was analyzed to find the loci responsible for the popping expansion volume. PMID- 17427036 TI - Inter-scan reproducibility of coronary calcium measurement using Multi Detector Row Computed Tomography (MDCT). AB - PURPOSE: To assess inter-scan reproducibility of coronary calcium measurements obtained from Multi Detector-Row CT (MDCT) images and to evaluate whether this reproducibility is affected by different measurement protocols, slice thickness, cardiovascular risk factors and/or technical variables. DESIGN: Cross-sectional study with repeated measurements. MATERIALS AND METHODS: The study population comprised 76 healthy women. Coronary calcium was assessed in these women twice in one session using 16-MDCT (Philips Mx 8000 IDT 16). Images were reconstructed with 1.5 mm slice thickness and 3.0 mm slice thickness. The 76 repeated scans were scored. The Agatston score, a volume measurement and a mass measurement were assessed. Reproducibility was determined by estimation of mean, absolute, relative difference, the weighted kappa value for agreement and the Intra-class correlation coefficient (ICCC). RESULTS: Fifty-five participants (72.4%) had a coronary calcification of more than zero in Agatston (1.5 mm slice thickness). The reproducibility of coronary calcium measurements between scans in terms of ranking was excellent with Intra-class correlation coefficients of >0.98, and kappa values above 0.80. The absolute difference in calcium score between scans increased with increasing calcium levels, indicating that measurement error increases with increasing calcium levels. However, no relation was found between the mean difference in scores and calcium levels, indicating that the increase in measurement error is likely to result in random misclassification in calcium score. Reproducibility results were similar for 1.5 mm slices and for 3.0 mm slices, and equal for Agatston, volume and mass measurements. CONCLUSION: Inter scan reproducibilility of measurement of coronary calcium using images from MDCT is excellent, irrespective of slice thickness and type of calcium parameter. PMID- 17427037 TI - Immunohistochemical detection of Langerhans cells in dental granulomas and radicular cysts. AB - OBJECTIVE: Dental granulomas (DGs) and radicular cysts (RCs) are chronic periapical lesions frequently involving the jaws. Langerhans cells (LCs) are dendritic cells responsible for the presentation of antigens to T lymphocytes. This study examined the expression of LCs in DG and RCs by immunohistochemical staining. STUDY DESIGN: Eighteen cases of DGs and 26 cases of RCs were analyzed using anti-CD1a marker. RESULTS: CD1a-labeled LCs were observed in 11.1% of DGs and in 69.2% of RCs, showing a significant correlation (P < 0.0001; Fisher's test). In DGs, LCs were only observed in granulation tissue, showing discrete immunostaining density. In RCs, LCs exhibited both a round and a dendritic shape in all epithelial layers. Although a correlation was observed between immunostaining density and epithelial thickness, as well as between immunostaining and inflammatory intensity, the differences were not significant in radicular cysts. CONCLUSION: Langerhans cells provide important insight into the immunopathogenesis of chronic periapical lesions. PMID- 17427038 TI - Tissue microarrays: robust molecular morphology tools. PMID- 17427039 TI - Delayed presentation of arteriovenous fistula 20 years after blunt renal trauma. AB - Delayed hematuria following blunt trauma is rare but can be significant. We describe an unusual delayed presentation of renal arteriovenous fistula following blunt trauma.A gentleman presented with hematuria and clot colic. Following initial investigations, renal arteriography showed an arteriovenous fistula in the mid-pole of the right kidney. The fistula was successfully occluded by super selective coil embolization. Although this case is an unusual presentation, AV fistulae must be ruled out in management of patients of hematuria. PMID- 17427040 TI - The Arabidopsis thaliana GSK3/Shaggy like kinase AtSK3-2 modulates floral cell expansion. AB - The GSK3/Shaggy family of serine/threonine protein kinases is involved in a series of biological processes in animals, plants and yeast [Charrier et al. (2002) Plant Physiol 130:577-590; Jope and Johnson (2004) Trends Biochem Sci 29:95-102; Li and Nam (2002) Science 295:1299-1301; Piao et al. (2001) Plant J 27:305-314]. In Arabidopsis thaliana, out of the 10 members of the GSK3/Shaggy like gene family (AtSKs), a biological function has been assigned to only 1 member (AtSK2-1) by mutation. In the present work, a study was undertaken to elucidate the function of AtSK3-2. We have generated mutated versions of the A. thaliana Shaggy-like kinase 3-2 (AtSK3-2), in which Lys(167) and Arg(178), respectively homologues to Lys(85) and Arg(96) of the mammal GSK3beta, were modified into Ala by site-directed mutagenesis. In vitro kinase activity assays of the mutated recombinant protein AtSK3-2-R178A showed that the "primed activity" of the mutated kinase was reduced by 90% while the "non-primed" activity was only 20% reduced compared to the wild-type protein kinase. However, the mutant protein AtSK3-2-K167A showed no activity. Arabidopsis transgenic lines over-expressing AtSK3-2-R178A displayed smaller floral organs, namely pedicels, sepals and petals. Conversely, over-expression of both the wild-type AtSK3-2 protein and the AtSK3-2-K167A mutated version, displayed no altered morphogenesis. Scanning electron microscopic analyses of the AtSK3-2-R178A transgenic plants clearly showed a reduced cell size in flower organs, in which quantitative RT-PCR expression analyses of cell wall expansion enzymes showed reduced transcript levels of three xyloglucan endotransglycosylases (XET), namely XTH22 (TCH4), XTH23 (XTR6) and XTH30 (XTR4). Our data show that AtSK3-2 plays an important role in the control of cell elongation in flower development. PMID- 17427041 TI - Identification of cellulose synthase AtCesA7 (IRX3) in vivo phosphorylation sites -a potential role in regulating protein degradation. AB - Cellulose is central to plant development and is synthesised at the plasma membrane by an organised protein complex that contains three different cellulose synthase proteins. The ordered assembly of these three catalytic subunits is essential for normal cellulose synthesis. The way in which the relative levels of these three proteins are regulated within the cell is currently unknown. In this work it is shown that one of the cellulose synthases essential for secondary cell wall cellulose synthesis in Arabidopsis thaliana, AtCesA7, is phosphorylated in vivo. Analysis of in vivo phosphorylation sites by mass spectrometry reveals that two serine residues are phosphorylated. These residues occur in a region of hyper variability between the cellulose synthase catalytic subunits. The region of the protein containing these phosphorylation sites can be phosphorylated by a plant extract in vitro. Incubation of this region with plant extracts results in its degradation via a proteasome dependant pathway. Full length endogenous CesA7 is also degraded via a proteasome dependant pathway in whole plant extracts. This data suggests that phosphorylation of the catalytic subunits may target them for degradation via a proteasome dependant pathway. This is a possible mechanism by which plants regulate the relative levels of the three proteins whose specific interaction are required to form an active cellulose synthase complex. PMID- 17427042 TI - [Are ICD-11 and DSM-V Necessary?]. PMID- 17427043 TI - [Clinical standing of valproate treatment of bipolar disorders]. AB - During recent years valproate has been established as a cornerstone for the drug treatment of bipolar disorder. In Germany, valproate was licensed both for the treatment of acute mania and for maintenance treatment in summer 2005. At this occasion, this review summarises the scientific evidence and clinical experience of well-known experts with valproate-treatment. It was concluded that valproate will continue to be of high clinical significance despite the recent increase of treatment alternatives, both in monotherapy and combination treatment of acute mania, mixed states and maintenance treatment. PMID- 17427044 TI - [Improvements in motor and non-motor symptoms in parkinson patients under ropinirole therapy]. AB - Ropinirole is a non-ergoline selective D2 dopamine agonist. Its efficacy and safety has been established in several controlled double-blind studies in patients with early and advanced Parkinson's disease. It is assumed that the improvement in the activities of daily living under ropinirole is not only due to the improved motor symptoms but also due to the improvement of non-motor symptoms like symptoms of mood and anxiety. The objective of this post marketing surveillance study was to show that under the conditions of the daily routine in the neurologic practice ropinirole may not only improve motor symptoms, the activity of daily living and complications of the treatment (dystonia, dyskinesia) but also alleviate symptoms of depression and anxiety. A total of 110 neurological practices enrolled 327 patients in early and advanced stages of the disease (139 females, 188-males; mean age: 67 years). They were treated with ropinirole as monotherapy and as adjunctive therapy with l-dopa over a period of 12 - 14 weeks. Selected symptoms of the Unified Parkinson's Disease Rating Scale (UPDRS) part II-IV and symptoms of depression and anxiety were rated by the clinicians. Mood and functional impairment in job, family and social life were rated by the patients using selected items of the Beck Depression Inventory and the Sheehan Disability Scale (SDS). The different subtypes, i. e. the akinetic rigid, tremor-dominant and the mixed subtype, are described separately. The total UPDRS score at baseline was similar for all three subtypes and there was also a similar improvement in the three groups under ropinirole. Both according to self rating and to clinician rating the symptoms of depression and anxiety at baseline were more severe in the akinetic-rigid and the mixed subtype compared to the tremor-dominant subtype. The symptoms considerably improved and were reduced by 48 % under therapy with ropinirole. Adverse events were reported by 7.7 % of the patients. The surveillance study has shown that ropinirole may improve not only motor symptoms, activities of daily living and complications of treatment but also symptoms of mood and anxiety. PMID- 17427045 TI - [Consultation- and liaisonpsychiatry]. PMID- 17427046 TI - Continuing education self-study program. PMID- 17427047 TI - Pragmatics and adult language disorders. PMID- 17427049 TI - Pragmatics and adult language disorders: past achievements and future directions. AB - In this article, the current state of our knowledge of pragmatic disorders in adults with language impairment is assessed. A brief historical background of clinical pragmatics is presented, and the place of adult language pathology within the development of this field is discussed. A comprehensive review is undertaken of pragmatic deficits in adults with language impairments of diverse etiologies. Specifically, pragmatic deficits are examined in adults with left hemisphere damage, often resulting in aphasia, and in adults with right hemisphere damage, traumatic brain injury, schizophrenia, and neurodegenerative disorders (principally, Alzheimer's disease). Although many pragmatic phenomena have been examined in these clinical populations, studies have also tended to neglect important areas of pragmatic functioning in adults with these disorders. Several such areas are identified within a wider discussion of how researchers and clinicians can best pursue future investigations of pragmatics in adults with language impairment. PMID- 17427050 TI - Cognitive pragmatics of language disorders in adults. AB - Cognitive pragmatics is the study of the mental structures and processes involved in the use of language in communicative contexts. Paradigms of cognitive psychology (off-line and on-line) have been applied to the study of the abilities to go beyond the literal (inference) and derive meaning in relation to context (e.g., metaphor and sarcasm). These pragmatic functions have been examined for the involvement of processes of meaning activation, embellishment, and revision. Clinical investigators have explored abilities and deficits in acquired aphasia, right hemisphere dysfunction, and closed head injury. This article reviews and provides some analysis of clinical studies that are consistent with the themes constituting cognitive pragmatics. PMID- 17427051 TI - Management of discourse deficits following traumatic brain injury: progress, caveats, and needs. AB - Currently, there is substantial evidence to support the assessment of communication, following traumatic brain injury (TBI), beyond what is included in standardized aphasia or child language batteries. The sensitivity of discourse analyses for delineating subtle cognitive-communicative deficits is well established in the research literature. A variety of useful monologic and conversational discourse measures have been identified including productivity, efficiency, content accuracy and organization, story grammar and coherence, and topic management. Disruption of discourse may persist for years after TBI and appears to have a negative impact on quality of life. Although discourse analyses have great potential for clinical application for individuals with TBI, their widespread use has been limited by time and training constraints. A further limitation is that there is no empirical evidence to guide the treatment of discourse deficits. There is need for the development and investigation of theoretically based interventions. PMID- 17427052 TI - Using conversation analysis to assess and treat people with aphasia. AB - This article gives an overview of the application to aphasia of conversation analysis (CA), a qualitative methodology for the analysis of recorded, naturally occurring talk produced in everyday human interaction. CA, like pragmatics, considers language use in context, but it differs from other analytical frameworks because the clinician is not making interpretations about how an aspect of language should be coded or judging whether an utterance is successful or adequate in terms of communication. We first outline the CA methodology before discussing its application to the assessment of aphasia, principally through the use of two published assessment tools. We then move on to illustrate applications of CA in the field of aphasia therapy by discussing two single case study interventions. Key conversation behaviors are illustrated with transcripts from interactions recorded by the person with aphasia and the person's habitual conversation partner in the home environment. Finally, we explore the implications of using CA as a tool for assessment and treatment in aphasia. PMID- 17427053 TI - Pragmatics in discourse performance: insights from aphasiology. AB - This article examines the preservation of pragmatic abilities of individuals with aphasia, as manifested in the discourse they produce. The construct of coherence is used as a framework for understanding this pragmatic preservation. Discourse coherence is largely derived from the structure, selection, and highlighting of information expressed in a discourse. Personal narratives, as one type of discourse, represent an extended turn-in-conversation on a topic of personal relevance to the speaker, common in everyday life. As such, they provide a valuable source of information about a speaker's pragmatic ability. Examples of personal narratives told by individuals with aphasia are used to illustrate and discuss the means by which discourse coherence is achieved. These include a tightly structured temporal-causal event line, development of theme, and evaluation of information. Possible approaches to clinical assessment are considered, including use of global rating systems. PMID- 17427055 TI - External quality assurance in thrombosis and hemostasis: an international perspective. AB - External Quality Assurance in Thrombosis and Hemostasis (EQATH) was organized in 2005 as an international collaboration of external quality assurance (EQA) programs and organizations with a common interest in improving the quality of hemostasis testing. The goals include exchanging of information regarding program operations; exchanging split specimens among programs to determine if there are differences in practice; participating in value setting of standards; and providing outreach programs to locations in the world without EQA support of hemostasis testing in laboratories. The organization currently includes 11 EQA programs from 10 countries. A survey of program structure and function revealed variation in the size and structure among the programs. In general, the staffing levels paralleled the size and complexity of the program. The number of laboratory participants in the EQA programs ranged from 58 to 1700. The presentation of testing covered in the program (modules) ranged from a program with a single module of a single test, to programs with single modules of many different types of tests, to a program with 13 modules, each of which contains a limited scope of related tests. Participating laboratories were graded (pass/fail or out with consensus) by six of the EQA programs, whereas five programs report the results to the laboratory for self-evaluation. Of the 11 programs responding, seven have deemed status from an accrediting or licensing agency, and successful participation satisfied requirements for accreditation for the participating laboratory. This type of benchmarking activity and cooperative activity among EQA programs will lead to improvement of the programs. PMID- 17427056 TI - Emerging technologies in hemostasis diagnostics: a report from the Australasian Society of Thrombosis and Haemostasis Emerging Technologies Group. AB - Technology in hemostasis laboratories has evolved enormously during the last 30 years. Although many scientists and clinicians will remember the traditional tilt tube techniques to screen for coagulation abnormalities and to monitor anticoagulant therapy, the hemostasis laboratory today uses a variety of modern technologies. These include flow cytometry, chromogenic assays, molecular typing (e.g., polymerase chain reaction), immunologic assays (e.g., enzyme-linked immunosorbent assays), functional assays of specific coagulation proteins, and platelet function analyzers. Although these advances in technology have resulted in greater capability, productivity, sensitivity, specificity, and ultimately, improvement in the clinical care of patients, controversies and limitations remain. This article highlights new and emerging technologies in hemostasis and discusses whether they have improved or are likely to improve laboratory diagnostics by specifically addressing the following: (1) Can new technologies help predict likelihood of thrombosis recurrence? (2) Has an understanding of the role of A Disintegrin-like And Metalloprotease with Thrombo Spondin type 1 motifs (ADAMTS13) in microangiopathy resulted in improved diagnostic methods for this disorder? (3) Does thrombelastography allow better definition of bleeding risk than conventional hemostasis assays, especially in settings of acute hemostatic pathology? PMID- 17427057 TI - Emerging technologies and quality assurance in hemostasis: a review of findings from the Royal College of Pathologists of Australasia Quality Assurance Program. AB - Regular multilaboratory surveys of laboratories by the Royal College of Pathologists of Australasia Quality Assurance Program (QAP) have been conducted to assess proficiency in tests of hemostasis for the last 40 years. This article focuses primarily on specialized assays of hemostasis, for which surveys have been conducted for some 10 years. For von Willebrand disease (vWD) evaluations, a total of 47 plasma samples have been dispatched to survey participants, including representative samples from normal individuals plus all of the major vWD subtypes (i.e., types 1, 2A, 2B, 2M, 2N, and 3). These surveys have focused partly on the issue of diagnostic interpretive error rates associated with different assays and test panels. In this context, considerable improvement is seen when laboratories incorporate the vWF:collagen-binding assay into the test panel. Thrombophilia associated tests assessed by the program and discussed in this review include activated protein c resistance, lupus anticoagulant, and deficiencies of protein C, protein S, and antithrombin. Other tests briefly reviewed here include factor assays and inhibitors, D-dimer, and heparin/anti-Xa assays. Anticardiolipin antibody and anti-beta(2)-glycoprotein I antibody (aB(2)GPI) testing, assessed by the Immunology QAP, is also reviewed briefly, as are genetic tests associated with thrombophilic markers such as factor V Leiden and the prothrombin gene. PMID- 17427058 TI - Emerging technologies and quality assurance: the United Kingdom National External Quality Assessment Scheme perspective. AB - The pattern of tests employed and technologies developed within hemostasis laboratories has changed considerably within the last 10 years. These changes have presented challenges to external quality assessment (EQA) providers, including the United Kingdom National External Quality Assessment Scheme (NEQAS). EQA for point-of-care devices used for monitoring oral anticoagulant therapy has focused on provision of suitable material to assess performance of devices designed for capillary blood testing, and on education of a user group not usually trained in laboratory quality control procedures. Development of novel therapeutic agents for hemophilia has presented challenges regarding standardization of assays for monitoring treatment, whereas advances related to laboratory testing and automation have not always been accompanied by improved accuracy and precision. EQA provision has also been shown to be of value in molecular genetic screening tests for thrombophilia, and in highlighting standardization issues related to D-dimer measurement in the exclusion of deep vein thrombosis. The increasing prevalence of screening tests of global hemostasis, such as thrombin generation tests and thromboelastography, presents additional challenges to EQA providers in the attempt to standardize these new and potentially beneficial technologies. PMID- 17427059 TI - Quality assurance in hemostasis: the perspective from the College of American Pathologists proficiency testing program. AB - External quality assurance (EQA) is an important component of the total quality assurance program of a clinical hemostasis laboratory. The College of American Pathologists (CAP) helps meet this requirement by providing a proficiency testing program that evaluates a broad range of hemostasis methods and analytes. This article reviews the published experience of the CAP proficiency testing program in hemostasis. The purpose is to formulate general conclusions about the benefits of EQA. Between 1963 and 2006, the performance characteristics of a variety of tests have been evaluated, including the prothrombin time, activated partial thromboplastin time, coagulation factor activity assays (e.g., fibrinogen, factor [F] VIII, FIX, FXI), von Willebrand factor assays, unfractionated heparin monitoring, lupus anticoagulant testing, and platelet function. Based on the results of these evaluations, the major benefits of EQA are to (1) enhance patient care and safety through improved laboratory testing; (2) characterize test accuracy and precision across multiple methods; (3) correlate specific method variables with accuracy and precision; (4) identify interfering substances and quantify their effects across multiple methods; (5) identify clinical laboratories that are at risk for poor performance so that their performance can improve; and (6) satisfy accreditation and regulatory requirements. PMID- 17427060 TI - External quality assessment schemes in coagulation in Germany: between regulatory bodies and patient outcome. AB - In the early 1980s, the first external quality assessment schemes (EQAS) regarding parameters of the coagulation laboratory were established in the daily routine of German laboratories. At present, the EQAS performed by INSTAND offers a wide range of global and single parameters of thrombosis and hemostasis. Only the participation in prothrombin time and activated partial thromboplastin time exercises is mandatory. However, devices applied for patient self-management of oral anticoagulation are not included. This article provides an overview of the EQA activities of INSTAND regarding different coagulation parameters. Problems in acquiring adequate sample material and strategies in establishing acceptability ranges are also discussed. PMID- 17427061 TI - External quality assessment scheme for hemostasis in India. AB - Regular participation in an external quality assessment scheme (EQAS) is critical for ensuring acceptable laboratory performance. However, participation in such programs is uncommon for laboratories performing tests of hemostasis in developing countries. There are several reasons, including lack of awareness of its significance, absence of locally administered and easily accessible programs, and costs associated with some of the international schemes. To address this problem, we initiated an EQAS for hemostasis in India in the year 2000. This initially was limited to approximately 25 laboratories associated with the chapters of the Hemophilia Federation (India), with samples and analysis of results supported by United Kingdom National External Quality Assessment Scheme. This was converted to a national program in 2003, in association with the Indian Society of Haematology and Transfusion Medicine. Local manufacture of survey samples began in 2004, along with analysis of results. Currently, more than 100 laboratories are registered in the program. They receive samples three times a year for the following tests: prothrombin time, activated partial thromboplastin time, and factor assays. Some surveys also include samples for fibrinogen and von Willebrand factor assays. In recent surveys, 60 to 95% of laboratories had their clotting times and 57 to 77% of laboratories had their factor assays within consensus. The program has helped identify causes of unacceptable performance. The challenges ahead are to increase participation, improve reporting of results, and provide individualized support to laboratories to improve performance when necessary. PMID- 17427062 TI - Platelet function testing: quality assurance. AB - Platelet function tests are widely used for the diagnosis of platelet disorders. In recent years there has been increasing interest in the use of platelet function tests to monitor antiplatelet drug therapy. Quality assurance is important to optimize the performance of laboratory assays but it has not been widely applied to platelet function tests. This deficiency likely reflects the need to use freshly collected blood samples for platelet function tests, and the complex, time-consuming nature of some assays such as aggregation studies. Platelet function testing lacks guidelines, is poorly standardized between laboratories, and rarely is evaluated by internal and external quality assurance exercises. The sensitivity, specificity, and diagnostic utility of some newer, simplified assays of platelet function have been evaluated in a range of clinical settings but corresponding quality assurance data for many established as well as emerging platelet function assays are lacking. Quality assurance issues relevant to testing platelet function are reviewed in this article, with a focus on their application to established and to new and emerging tests. PMID- 17427063 TI - International biological standards for coagulation factors and inhibitors. AB - The use of international biological standards during the last 30 years has proved extremely successful in promoting global harmonization of estimates between laboratories and methods. Experience has led to the identification of physical criteria essential for standards to be suitable for long-term use. High precision of liquid filling coupled with low residual moisture and oxygen and the use of sealed glass ampoules have been found consistent with homogeneous and stable International Standards (ISs). Most plasma coagulation factors and inhibitors are calibrated in International Units (IU), which are defined as the amount of analyte in 1 mL of normal pooled plasma. Adoption of the IU has provided clarity in the definition of normal and abnormal states and has facilitated dose calculation for replacement therapy. The assay of like-versus-like materials (e.g., concentrate versus concentrate) has been found to improve interlaboratory agreement and there are now both plasma and concentrate ISs available for many coagulation factors and inhibitors. Studies into the assay of recombinant factor VIII have indicated that additional measures, such as modifications to assay methodology, are necessary to reduce interlaboratory variability. This experience may prove valuable in the future, when we have to deal increasingly with the challenges to standardization associated with the products of bioengineering. PMID- 17427064 TI - Standardization, regulation, quality assurance and emerging technologies in hemostasis: issues, controversies, benefits, and limitations. AB - There are many benefits to the overall process of standardization for tests of hemostasis and thrombosis. Nevertheless, it should also be recognized there are several specific problems and limitations to this process, as highlighted in this review. Some of the issues are formidable, but it is hoped that they are not insurmountable. Sometimes, clinical pressures drive diagnostic test processes before they are formally proven to be of clinical value. Such clinical pressures drive diagnostic changes in the hemostasis laboratory, including the incorporation of new and emerging technologies, which in turn drives the need for evolving and effective external quality assurance. Incorporated into the diagnostic or test-performance process are a large number of organizations involved in driving standardization, with the ultimate intention of improving diagnostics, but this process will also have unintended potential for adverse outcomes. Although this review notes many of the benefits to the process, it focuses primarily on those negative factors that are often less obvious but still require attention and process review. PMID- 17427065 TI - Endoscopic ultrasound versus endoscopic retrograde cholangiography for patients with intermediate probability of bile duct stones: a randomized trial comparing two management strategies. AB - BACKGROUND AND STUDY AIMS: Diagnostic endoscopic retrograde cholangiography (ERC) is being replaced by endoscopic ultrasonography (EUS) in patients with suspected bile duct stones. The assumption that such an approach is advantageous, however, has never been tested in a randomized trial. PATIENTS AND METHODS: 100 patients with intermediate probability of bile duct stones were randomly allocated to EUS or ERC. Two patients in the ERC group were excluded; the remaining 98 patients received the allocated intervention and were entered into the analysis (EUS, 50 patients; ERC, 48 patients). Detected stones were removed endoscopically; patients without stones were followed for 1 year. The primary end point was the proportion of patients with a negative outcome, related to either endoscopic procedures (complications) or to false-negative diagnosis of stones. Investigators assessing the negative outcomes were not blinded to group assignment. The secondary end point was the total number of endoscopic procedures (EUS and ERC) performed in each group to diagnose and treat stones. RESULTS: Bile duct stone prevalence was 28% and 25% in the EUS and ERC groups, respectively (P > 0.05). In the EUS group, 71 endoscopic procedures were performed, and 63 in the ERC group (mean per patient, 1.42 +/- 0.76, and 1.31 +/- 0.55, respectively; P > 0.05). In the EUS group, these included 49 successful and one failed initial EUS, 15 ERCs for bile duct stone treatment, and six procedures required during follow up. In the ERC group there were 36 successful and 12 failed initial ERCs, 13 repeat procedures (EUS or ERC) performed after failed or equivocal initial ERC, and two procedures during follow-up. Five patients in the EUS group (10%, 95% CI 4-22) and 19 patients in the ERC group (40%, 95% CI 27-54) experienced a negative outcome (P < 0.001). No difference was observed when only moderate to severe complications were considered (6%, 95% CI 1-17, and 10%, 95% CI 4-23, respectively). CONCLUSIONS: In patients with intermediate probability of bile duct stones, the management strategy based on EUS (with selective ERC in patients with confirmed stones) is safer and not associated with an excess of endoscopic procedures compared with a strategy based on ERC alone. PMID- 17427067 TI - Endoscopic gallbladder drainage of patients with acute cholecystitis. AB - BACKGROUND AND STUDY AIM: The standard treatment for acute cholecystitis is early laparoscopic cholecystectomy. In cases of increased operative risk surgery may be postponed or rejected, and instead alternative methods, such as percutaneous or endoscopic drainage, may be attempted. This paper is a retrospective assessment of our results with endoscopic gallbladder drainage (EGBD) by means of endoscopic retrograde cholangiopancreatography (ERCP). PATIENTS AND METHODS: Over a 9-year period, data from all patients who underwent attempted EGBD, primarily referred for diagnosis and treatment of cholestasis by ERCP, were analyzed. EGBD was attempted in those patients who had concomitant acute cholecystitis. RESULTS: EGBD was successful in 24 of 34 patients with acute cholecystitis (70.6%). The success rate rose from 50% during the first 4-year period to 89% during the subsequent 5-year period. Of the 24 patients in whom EBGD had been successful, 21 showed clinical improvement and 10 were finally managed nonoperatively. Of the 21 clinically improved patients, 14 underwent elective surgery a median of 24 days later. Of the 10 patients in whom EGBD failed nine underwent surgery four of whom required surgery within one week. CONCLUSION: Treatment of acute cholecystitis in patients could be done successfully by EGBD, and in 70% of cases cholecystectomy could then be carried out on an elective basis rather than as emergency surgery. Some patients could be treated with EGBD alone. Future prospective trials will clarify the role of EGBD in patients with acute cholecystitis. PMID- 17427068 TI - Publication bias does not play a role in the reporting of the results of endoscopic ultrasound staging of upper gastrointestinal cancers. AB - BACKGROUND AND STUDY AIM: An overestimation of the accuracy of endoscopic ultrasound (EUS) results in rectal cancer staging has been reported recently, which was found to be caused by the selective reporting of more positive results. In this study, we assessed whether publication bias was also present in the reporting of EUS staging results in upper gastrointestinal cancer. METHODS: A Medline literature search was performed. English-language articles containing information on the accuracy of EUS for T staging and/or N staging of esophageal, gastric, and pancreatic cancer were included. Articles published in abstract form only, case reports, and reviews were excluded. Studies reporting EUS results of patients who had undergone preoperative radiation and/or chemotherapy were also excluded. EUS results were plotted against numbers of patients, year of publication, journal type, and journal impact factor. RESULTS: The plots of the numbers of patients against accuracies for T stage and N stage and the statistical analyses showed no evidence of publication bias with regard to upper gastrointestinal cancer. The reported accuracy of EUS for the T stage of esophageal, gastric, and pancreatic cancer declined slightly over the years, but this was statistically significant only in the case of esophageal cancer (P = 0.01). No statistically significant correlations were found for N staging for any of the three types of cancer. In addition, no correlations were found between EUS results and journal type or journal impact factor. CONCLUSION: No evidence was found for the selective reporting of more positive EUS results for esophageal, gastric, and pancreatic cancer staging, which suggests that publication bias was not present. PMID- 17427069 TI - Predicting outcomes and complications of percutaneous endoscopic gastrostomy. AB - BACKGROUND AND STUDY AIMS: Percutaneous endoscopic gastrostomy (PEG) is the preferred route for long-term enteral feeding. Our aims were to prospectively evaluate the outcome ("PEG status") and complications of PEG and to determine whether these can be predicted by patients' baseline characteristics. PATIENTS AND METHODS: We conducted a prospective study in two tertiary hospitals between August 2003 and January 2005, enrolling all patients who were undergoing PEG placement. We completed a questionnaire with details of demographic data, diagnosis, indication for PEG, Charlson's co-morbidity index, Barthel's index, laboratory tests, complications, and date and cause of death. Patients were followed at scheduled appointments. Univariate and multivariate analyses were performed. RESULTS: 168 patients (48% male, 52% female; mean age +/- standard deviation 74 +/- 16 years) underwent PEG using the pull technique. The main indication was neurogenic dysphagia (156 patients, 92.9%). Although most indications were appropriate, in half the cases these were established too late. There were no procedure-related deaths. Major complications occurred in four patients (2.4%); minor complications occurred in 52 patients (31%). No single variable could predict complications. Fifteen patients (9%) had the PEG removed. No single variable was independently associated with PEG removal. The mortality was 6.5% at 30 days, 17.3% at 90 days and 33.9% at 1 year. The C-reactive protein was the only predictive factor of early mortality (< or = 30 days), and Charlson's co-morbidity index was the only predictive factor of late mortality (> 30 days). CONCLUSIONS: PEG placement is an easy and safe procedure, although it is often requested too late. No single variable could predict complications or PEG removal. C-reactive protein was found to be predictive of early mortality and Charlson's index was predictive of late mortality. PMID- 17427070 TI - Analysis of symptoms and endoscopic findings in 117 patients with histological diagnoses of eosinophilic esophagitis. AB - BACKGROUND AND STUDY AIMS: Although eosinophilic esophagitis has been increasingly diagnosed over recent years, little is known about this disease. In this study, symptoms, accompanying allergic disorders, and endoscopic findings in 117 patients with eosinophilic esophagitis were analyzed retrospectively. PATIENTS AND METHODS: The physicians who had treated the 117 patients (mean age 42.2 years; 9 children, 108 adults; male patients 71.8%) with the histological diagnosis of eosinophilic esophagitis were asked to provide data on symptoms, accompanying allergic disorders, and endoscopic findings. RESULTS: In 82.2% of the patients symptoms appeared in adulthood, predominantly between the ages of 21 and 30 years. The average duration of symptoms until final diagnosis of eosinophilic esophagitis was 4.2 years (range 0-44 years). The most frequent symptom was dysphagia (70.1%), followed by heartburn (47%), chest pain (29%), epigastric pain (29%), and a combination of dysphagia and heartburn (29%). Allergic disorders were seen in 48.7% of our patients. The most frequent endoscopic findings were stipple-like exudates (25.6%), linear fissures (25.6%), and reddening (25.6%), followed by rings (18.8%) and strictures (16.2%) of the esophagus. The esophageal mucosa was regarded as "normal" in 24.8% of the patients. CONCLUSION: Dysphagia in the second or third decade of life may suggest eosinophilic esophagitis. Symptoms of eosinophilic esophagitis may be indistinguishable from those of gastroesophageal reflux disease. The endoscopic appearance is not specific. Biopsies taken from multiple locations in the esophageal mucosa are essential for diagnosis of eosinophilic esophagitis. PMID- 17427071 TI - In-vivo confocal real-time mini-microscopy in animal models of human inflammatory and neoplastic diseases. AB - BACKGROUND AND STUDY AIMS: Although various improvements in tissue imaging modalities have recently been achieved, in-vivo molecular and subsurface imaging in the field of gastroenterology remains a technical challenge. In this study we evaluated a newly developed, handheld, miniaturized confocal laser microscopy probe for real-time in-vivo molecular and subsurface imaging in rodent models of human disease. MATERIALS AND METHODS: The minimicroscope uses a 488-nm, single line laser for fluorophore excitation. The optical slice thickness is 7 microm, the lateral resolution 0.7 microm. The range of the z-axis is 0-250 microm below the tissue surface. Imaging was performed using different fluorescent staining protocols; 5-carboxyfluorescein-labeled octreotate was synthesized for targeted molecular imaging. RESULTS: Cellular and subcellular details of the gastrointestinal tract could be visualized in vivo at high resolution. Confocal real-time microscopy allowed in-vivo identification of tumor vessels and liver metastases, as well as diagnosis of focal hepatic inflammation, necrosis, and associated perfusion anomalies. Somatostatin-receptor targeting permitted in-vivo molecular staining of AR42-J-induced carcinoma and pancreatic islet cells. CONCLUSIONS: Confocal mini-microscopy allows rapid in-vivo molecular and subsurface imaging of normal and pathological tissue in the gastrointestinal tract at high resolution. Because this technology is applicable to humans, it might impact on future in-vivo microsocpic and molecular diagnosis of diseases such as cancer and inflammation. PMID- 17427072 TI - EUS-first versus ERC-first for patients with intermediate probability of bile duct stones: is one strategy superior to the other? PMID- 17427073 TI - Management of acute cholecystitis and the surgeon's dilemma: the gun shoots both directions. PMID- 17427074 TI - Argon plasma coagulation in chronic radiation proctitis. AB - Chronic radiation proctitis is a relatively common late complication of pelvic irradiation. The main symptoms are diarrhea, urgency, tenesmus, and rectal bleeding. While mild cases may settle spontaneously over some months, severe hemorrhagic radiation proctitis may require repeated blood transfusions and is difficult to treat with medical therapy. Argon plasma coagulation (APC) is a noncontact thermal coagulation technique which can be applied endoscopically. A probe passed through the scope delivers a field of argon gas to the mucosal surface where it is ionized by a high voltage filament, resulting in superficial mucosal heating and coagulation of friable blood vessels. The technique reduces rectal bleeding in 80%-90% of cases, and may improve the other troublesome symptoms of diarrhea and urgency. APC is probably less effective in very severe cases of hemorrhagic radiation proctitis; in these cases topical formalin or a combination of APC and topical formalin can be useful. Overall, APC has proved to be a safe and well tolerated technique. PMID- 17427075 TI - In-vivo microvascular imaging of early squamous-cell cancer of the esophagus by confocal laser endomicroscopy. AB - We present the case of a 63-year-old man with an early esophageal squamous-cell cancer that was visualized and analyzed by fluorescein-aided confocal laser endomicroscopic imaging. Our findings indicate that endomicroscopy is readily able to provide images of microvessels and to differentiate between normal and abnormal tumor vessels. Endomicroscopy therefore has the potential to characterize microvascular changes in early squamous-cell cancer, suggesting that this technique might have major implications for optimizing treatment strategies in modern endoscopy. PMID- 17427076 TI - Detection of tissue abnormalities in normal mucosa surrounding colorectal cancer using an endocytoscopy system. AB - The endocytoscopy system is a novel diagnostic technique that provides extremely high-magnification imaging of the gastrointestinal mucosa. We are currently using a prototype Olympus endocytoscope for clinical research in gastrointestinal tumors. In one surgical specimen obtained after resection of a cancer of the transverse colon, focal abnormalities of colonic glands were detected 7 cm away from the primary tumor, within macroscopically normal mucosa. Our finding, which was confirmed by histopathological examination, suggests the need for further clinical investigation to assess whether endocytoscopy is able to identify premalignant changes in apparently normal mucosa. This could potentially be useful for accurate evaluation before planning minimally invasive therapy. PMID- 17427077 TI - Can the triclip be successfully applied in clinical practice? PMID- 17427079 TI - Endoscopic treatment of bleeding esophageal varices. PMID- 17427082 TI - Elastosonography in malignant rectal disease: preliminary data. PMID- 17427083 TI - Endoscopic dilation in eosinophilic esophagitis: a treatment strategy associated with a high risk of perforation. PMID- 17427085 TI - Early endoscopic sphincterotomy in acute cholecystitis in elderly patients with high surgical risk. PMID- 17427088 TI - [Interdisciplinary oncology using pancreatic cancer as an example]. PMID- 17427089 TI - [Surgical treatment of pancreatic cancer]. PMID- 17427090 TI - [Adjuvant treatment of pancreatic cancer]. PMID- 17427091 TI - [The role of radiochemotherapy in multimodal therapy of pancreatic cancer]. PMID- 17427092 TI - [Palliative treatment of pancreatic cancer]. PMID- 17427093 TI - [Molecular-based treatment concepts in advanced pancreatic cancer]. PMID- 17427094 TI - [Hamartoma of the lung]. PMID- 17427095 TI - [Geriatric assessment]. PMID- 17427096 TI - [CME and continued education]. PMID- 17427097 TI - [Body images of male patients with eating disorders]. AB - Ideals of male attractiveness have changed considerably. The ideal male body at present is characterized by low body fat and pronounced muscles. Similar to what has been found for women, these normative societal conceptions should influence the pathology of men with eating disorders. In the present study, men with and without eating disorders are compared regarding body satisfaction and body perception. Both questionnaire data and a computer assisted approach are applied. Men with bulimia nervosa wish to have a body with less fat whereas men with anorexia do not wish for a bigger body although they are seriously underweight. Men in all groups wish to have more muscles. Men with and without an eating disorder do not differ in this respect. The wish for less body fat and more muscles is associated with body dissatisfaction in men. Treatment of men with eating disorders should focus on men's body images similar to how it is conceptualized in treatments for women with eating disorders. Different from women, a body image focused approach for men should emphasize the meaning of muscularity. PMID- 17427098 TI - [The parent-child relationship inventory for children: background and first results]. AB - A questionnaire on representations of the parent-child relationship in preadolescent children is presented. At present, no standardised measure is available for this age group which assesses economically but still differentiated how younger children perceive the relationship with their parents. The original version of the Parent-Child Relationship Inventory for Children (PI-C, in German: FEB-K) was given to a sample of 371 fourth-grade students of different elementary schools in Hamburg. Factor- and item analysis led to a 22-item questionnaire with the dimensions Care, Control, Lack of Limitations, Confidence, and Conflict/Rejection. A total of 53.8 % of variance is explained by the 5-factor solution. Based on these factors, distinct groups of the perceived parent-child relationship emerged in cluster analysis which were associated with children's eating behaviour. The PI-C is proposed for research on the parent-child relationship in clinical as well as non-clinical samples and for the use as diagnostic instrument in clinical practice. PMID- 17427099 TI - [Effectiveness of dialectical behavior therapy for patients with borderline personality disorder in the long-term course--a 30-month-follow-up after inpatient treatment]. AB - BACKGROUND: The beneficial effects of Dialectical Behavior Therapy (DBT) for patients with borderline personality disorder (BPD) are well established. However, it is not well known whether this type of treatment relieves symptoms and signs of BPD in the long-term course thereafter and whether the results of DBT are transferable for patients with high comorbidity. METHODS: We conducted a follow-up examination of 50 consecutive inpatients with BPD as defined by DSM-IV. The patients were examined at admission, at discharge and 15 and 30 months after discharge. For the clinical diagnosis and to survey psychopathology we used the Structured Clinical Interview for DSM-IV (SCID), the Global Assessment of Functioning (GAF) and several self-rating-instruments. RESULTS: Compared to admission 30 months after discharge we observed the following results: A significant number of patients did not meet the DSM-IV criteria for BPD anymore, comorbidity (particularly mood disorders, drug or alcohol abuse/dependence and eating disorders) was reduced, psychosocial functioning was improved and general and BPD-typical symptoms were relieved. CONCLUSION: Our findings support the efficacy of DBT in an inpatient setting and show that the achieved success of therapy is stable for a prolonged period of time. Patients with high comorbidity seem to profit from DBT as well. PMID- 17427100 TI - [Mindfulness training as a psychotherapeutic tool. Clarification of concept, clinical application and current state of empirical research]. AB - With mindfulness-based interventions becoming increasingly popular this review outlines: 1. How mindfulness is defined and conceptualized, 2. How mindfulness is applied in psychotherapy, 3. How effective mindfulness-based interventions are and 4. How the effectiveness of these methods might be explained. Findings show that mindfulness can be seen as a multidimensional concept which is the core or an important component of several psychotherapeutical methods that have been shown to be effective in the treatment of different psychological problems and disorders. With regard to possible mechanisms of change it is argued that enhancing patients' skills to accept and tolerate aversive experiences can be seen as a specific and important effect of these approaches. Future research should try to clarify for whom these skills are particularly important, and how these skills can be taught to patients who are less open for mindfulness-based approaches. PMID- 17427101 TI - [Screening]. PMID- 17427102 TI - Diagnostic management of benign and malignant pheochromocytoma. AB - As rare and thus often overlooked hormone-secreting tumors, pheochromocytomas pose a particular diagnostic challenge. Difficulties involve biochemical confirmation, localizing, and detection of malignancy. Measurement of free plasma metanephrines, genetic testing and specific imaging procedures--such as MIBG and octreotide scintigraphy or fluorodopamine PET--represent a considerable progress, and the management of benign pheochromocytomas has become very effective. However, a comparable improvement in the prognosis of malignant chromaffin cell tumors, which occur in approximately 10-15% of all cases, has not yet been achieved. Here, telomerase catalytic subunit (hTERT) activity and heat shock protein 90 expression could serve both as molecular markers allowing an earlier diagnosis of malignancy and as therapeutic targets. Familial syndromes should be considered both in benign and malignant pheochromocytoma, and should be tested for prior to surgery in selected patient groups. PMID- 17427103 TI - Genetics and biology of pheochromocytoma. AB - The familial forms of pheochromocytoma have recently been demonstrated to be more frequent than believed in the past. The genes currently known to be responsible for tumor formation are RET, VHL, NF1, SDHB, SDHC and SDHD. Germline mutations of these genes increase the risk of developing pheochromocytomas and/or paragangliomas which variably associate with other neoplasms and characterize diverse clinical syndromes such as MEN 2, von Hippel-Lindau (VHL), and neurofibromatosis type 1 (NF 1), or the PGL syndromes, respectively. Although the pathogenesis of pheochromocytoma/paraganglioma formation is still largely unknown, studies of the familial forms have started to uncover some pathways that favor tumor formation, such as activation of tyrosine-kinase, induction of hypoxia-inducible factors, activation of the oncogene Ras or reduced apoptosis. These studies have also demonstrated that various gene mutations can differently affect the biological characteristics of pheochromocytoma: for example, while the tumors are mostly adrenergic (epinephrine secreting) and episodically secreting in MEN 2, they are mostly noradrenergic (norepinephrine secreting) and continuously secreting in VHL. Biological variability can also be observed in the PGL syndromes where tumors develop in the head and neck and are parasympathetic in origin and non-secreting, or in the thorax and the abdomen, where they are sympathetic in origin and catecholamine secreting. Genetic testing in patients with pheochromocytomas or paragangliomas is, at present, strongly recommended and is mandatory in young patients or in cases of multiple or recurrent tumors. The clinical picture and the biological characteristics of the tumor may suggest the priority of the genes to be tested first. PMID- 17427104 TI - Management of adrenal incidentalomas. PMID- 17427105 TI - Detecting and treating primary aldosteronism: primary aldosteronism. AB - Primary aldosteronism (PA) is the most common cause of mineralocorticoid hypertension. Different studies, using the plasma aldosterone concentration to plasma renin activity ratio (PAC/PRA) for the screening of patients with hypertension, have shown a marked increase in the detection rate of PA. Idiopathic bilateral adrenal hyperplasia (IHA) and aldosterone-producing adrenal adenoma (APA), are the leading causes of primary aldosteronism. Glucocorticoid remediable aldosteronism (GRA), also called familial hyperaldosteronism type I, familial hyperaldosteronism type II and carcinomas are rare causes of PA. Patients with hypertension and hypokalemia, those with a family history of hypertension and stroke at an early age, or patients with medication-resistant hypertension should be screened for PA using the PAC/PRA ratio. If a high ratio is found, a sodium loading test or a captopril test is warranted to confirm the diagnosis. Adrenal gland imaging is important in subtype differentiation (APA vs IHA). Adrenal venous sampling should be used when other tests prove inconclusive. Genetic testing has facilitated detection of GRA. Surgery is considered the treatment of choice for patients with APA, while bilateral hyperplasia subtypes are treated medically. Normalization of aldosterone levels or aldosterone receptor blockade are necessary to prevent the morbidity and mortality associated with hypertension, hypokalemia, and cardiovascular damage. PMID- 17427106 TI - Insights into the molecular biology of adrenocortical tumors. PMID- 17427107 TI - Seeking the factors predisposing to local skin inflammatory state development in children with type 1 diabetes (T1DM) treated with continuous subcutaneous insulin infusion (CSII). AB - The often CSII treatment complication is local skin infection. The aim of the study was to analyze chosen factors predisposing to this complication. MATERIAL AND METHODS: We observed 40 children aged 1.9-15.6, suffering from diabetes for 0.1-12 and treated by CSII for 0.01-4.4 years in whom HbA1c, BMI, injection site and catheter insertion duration, catheter colonization, skin flora and Staphylococcus aureus carrier state were analyzed. The catheter cultures were prepared with Maki method. The skin and nasal vestibule swab were taken to detect local flora. RESULTS: In the culture of 43 catheters (Maki method) a positive growth (>10 cfu) was detected in 9 (21%), homogeny culture of coagulase-negative staphylococci in 7 and mixed culture (both S.epidermidis and S.aureus) in two cases. Skin inflammation of the injection site was observed in a total of 10 children (25%), in two of whom catheter culture was positive. A statistically significant relation between the presence of bacteria in the catheter and on the skin around the injection site was found. Among the examined parameters, the relation between the catheter colonization and HbA1c, female sex and BMI were observed. CONCLUSIONS: Metabolic control, female sex and BMI influence the development of a skin inflammatory state in patients treated with CSII. S.aureus carrier state has no impact either on catheter colonization or the development of an infection. However, bacteria skin occurrences can predispose to catheter colonization by the strain as well as to developing an inflammation. PMID- 17427108 TI - Long-term improvement of metabolic control without increased risk of hypoglycaemia by intensive insulin regimens in type 1 diabetes patients treated in a regular clinical setting. AB - AIM: To evaluate if intensive insulin regimen with multiple daily injections (MDI) is successful for treating type 1 diabetes patients over a long period of time in a regular clinical setting. METHOD: This is a prospective, observational seven-year study. Fifty-nine (35 male) type 1 diabetic patients with bad metabolic control (HbA1c> or =9%), aged 31.9 years, range 18-47 were included in the present study. All of them had had at least 5 years of diabetes duration after diagnosis and showed negative responses of serum C-peptide to a standard breakfast. The main control variables are: Metabolic control measured by serum HbA1c values (values < 6.2 % was the treatment objective) and the frequency of hypoglycaemic episodes (episodes/patient-month). RESULTS: Significant decreases in mean+/-SD HbA1c values in this group of patients were observed from the first year of follow-up, with the mean values being: 7.5+/-1.5%, 7.2+/-1.8%, 7.6+/ 1.6%, 7.1+/-1.7%, 7+/-1.4, 6.6+/-1.6% and 6.8+/-1.4% for the first, second, third, fourth, fifth, sixth and seventh years of follow-up respectively. Sixteen %, 27.5%, 15.7%, 33.3%, 28.6%, 42% and 33% of the patients reached the treatment objective (HbA1c values<6.2%) for each year of follow-up. Throughout the study period the rate of severe hypoglycaemia (episodes/patient-year) was 0.32+/-0.2 which was not significantly different compared with the value of 0.28+/-0.1 observed the year before the study began. Similarly frequencies of mild/moderate hypoglycaemia episodes (episodes/patient-month) varies between 16.5+/-4 and 21.7+/-5, which are not significantly different from the value of 17.7+/-6 observed the year before the study began. CONCLUSION: Long-term improvement in metabolic control was observed in this group of type 1 diabetes patients with previous bad control, during treatment in a regular clinical setting. A considerable percentage of type 1 diabetic patients with MDI reached the treatment objective in every year of follow-up. Furthermore improvement in metabolic control is not associated with significantly increased frequency of hypoglycaemia episodes. PMID- 17427109 TI - Distribution of the components of the NCEP ATP III-defined metabolic syndrome in newly diagnosed diabetes and non-diabetes caucasian subjects; implications for metabolic syndrome prevention and treatment. AB - The study aimed at assessing the relationship between five components of the NCEP ATP III-defined metabolic syndrome (MS) in diabetes and non-diabetes MS subjects. METHODS: Prevalence of MS and its components was assessed in 1586 adult Caucasian individuals, without known diabetes. Oral glucose tolerance test (OGTT) with 75 g glucose according to WHO protocol was conducted in all subjects. RESULTS: MS was diagnosed in 419 persons (prevalence 26.4%), including 123 subjects with newly diagnosed diabetes. Diabetes subjects presented with higher plasma triglycerides and slightly more pronounced abdominal obesity as well as higher fasting plasma insulin, which in all reflected insulin resistance-related character of MS pathophysiology. However, the distribution of MS components was largely uneven in diabetes and non-diabetes subjects. Elevated plasma triglycerides and low HDL cholesterol were the most prevalent components. The least often found component was elevated fasting plasma glucose (FPG). Non-diabetes subjects with elevated FPG were more likely to present with three or four other components than with two of them only. In contrast, those with high plasma triglycerides, low plasma HDL cholesterol or increased waist circumference were all more likely to have two rather than three or four other MS components. Subjects with diabetes presented significantly more often with four or five MS components when compared with non diabetes subjects. CONCLUSIONS: The components of metabolic syndrome are not equally prevalent in Caucasian population. Lipid disorders are most often found and glucose intolerance is the least prevalent MS component. Glucose intolerance and diabetes occurred in a more complex setting, concomitantly with three or four other MS components, suggesting that diabetes is associated with more advanced stages of MS. These findings may have important implications for MS prevention and treatment. PMID- 17427110 TI - Glucose metabolism, insulin secretion and insulin sensitivity in juvenile hemochromatosis. A case report and review of the literature. AB - Among specific diabetes subtypes secondary to pancreatopathies, hereditary hemochromatosis is an inherited disorder of iron metabolism resulting in excessive iron overload and tissue damage in various organs. We here report the case of a man with the young-onset form of the disease and describe his glycaemic status before and during venesection therapy. A 25-year old man visited our clinic in Athens, Greece, with hypogonadotropic hypogonadism due to hereditary hemochromatosis. Genetic analysis revealed that he was suffering from the juvenile aggressive form and treatment was initiated with frequent phlebotomies in conjunction with androgen substitution. Within 18 months of therapy ferritin level was normalized and hypogonadism was fully restored. Despite severe iron overload, glucose tolerance remained normal during the various stages of the disease, although alterations in both insulin secretion and sensitivity were detected. Present data indicate that in juvenile hemochromatosis, the efficacy of the chelation therapy and probably the chronic interval required to restore normal iron concentration both play important roles in the formation of glucose metabolism characteristics. PMID- 17427111 TI - Pegvisomant treatment in gigantism caused by a growth hormone-secreting giant pituitary adenoma. AB - BACKGROUND: Gigantism is rare with the majority of cases caused by a growth hormone (GH)-secreting pituitary adenoma. Treatment options for GH-secreting pituitary adenomas have been widened with the availability of long-acting dopamine agonists, depot preparations of somatostatin analogues, and recently the GH receptor antagonist pegvisomant. CASE REPORT: A 23-year-old male patient presented with continuous increase in height during the past 6 years due to a GH secreting giant pituitary adenoma. Because of major intracranial extension and failure of octreotide treatment to shrink the tumour, the tumour was partially resected by a trans-frontal surgical approach. At immunohistochemistry, the tumour showed a marked expression of GH and a sparsely focal expression of prolactin. Somatostatin receptors (sst) 1-5 were not detected. Tumour tissue weakly expressed dopamine receptor type 2. The Gs alpha subunit was intact. Conversion from somatostatin analogue to pegvisomant normalized insulin-like growth-factor-I (IGF-I) levels and markedly improved glucose tolerance. CONCLUSION: Pegvisomant is a potent treatment option in patients with pituitary gigantism. In patients who do not respond to somatostatin analogues, knowledge of the SST receptor status may shorten the time to initiation of pegvisomant treatment. PMID- 17427112 TI - Association between glycosylated hemoglobin level and 16-year incidence of chronic kidney disease in type 1 diabetes. AB - CONTEXT: The incidence of recently defined outcome of chronic kidney disease (CKD) has not been widely reported in type 1 diabetes. OBJECTIVE: To examine the prospective association between baseline glycosylated hemoglobin levels and the 16-year incidence of CKD and end-stage renal disease (ESRD) in type 1 diabetes. DESIGN: Prospective cohort study of type 1 diabetes individuals. SETTING: Community based in southwestern Wisconsin. PARTICIPANTS: 547 younger-onset type 1 diabetes individuals who were free of CKD at baseline (1984-86). MAIN OUTCOME MEASURES: Development of CKD (defined as estimated glomerular filtration rate<60 ml/min/1.73 m(2) or ESRD [history of dialysis or renal transplantation]) over 16 year follow-up period, among individuals free of CKD at baseline. Alternate outcome was 16-year incident ESRD. RESULTS: After 16 years of follow-up, there were 158 cases of CKD and 37 cases of ESRD in our cohort. The 16-year cumulative incidence of CKD was 31.7 percent. Elevated glycosylated hemoglobin levels were associated with incident CKD and ESRD in separate models. Multivariable odds ratio (OR) [95% confidence intervals (CI)] comparing the highest quartile of glycosylated hemoglobin (11-15.3%) to the lowest quartile (6-8.6%) was 6.44 (3.61 11.51), p-trend<0.0001 for incident CKD and 21.87 (2.84-168.39), p-trend<0.0001 for ESRD. CONCLUSIONS: Higher baseline glycosylated hemoglobin levels are independently associated with incident CKD and ESRD, among individuals with type 1 diabetes. PMID- 17427113 TI - Endosonographic diagnosis of solid pancreatic tumors: a retrospective analysis from a tertiary referral center. AB - BACKGROUND: The sensitivity of endosonography for pancreatic tumors is good, but differentiation between malignant and benign lesions remains a challenge. We, therefore, analyzed the correlation between endosonographic findings and pancreatic carcinoma in a population with a high prevalence of chronic pancreatitis. METHODS: 171 endosonographic examinations were retrospectively evaluated using 22 dichotomous criteria. Final diagnosis was defined by the results of pancreas resection or by clinical follow-up (median: 41 months). A sum score was established after multivariate analysis. RESULTS: Final diagnosis was carcinoma, chronic pancreatitis, neuroendocrine tumor and normal pancreas in 67, 65, 9, and 38 subjects (prevalence 39 %, 38 %, 5 %, 22 % respectively) After multivariate analysis, carcinoma was significantly correlated with six endosonographic criteria and age, which resulted in the following score (yes = 1, no = 0): (dilated pancreatic duct) + (vascular infiltration) + (suspicious lymph nodes) + (tumor edge with pseudopodia-like extensions) + (age > 50 years) - (increased pancreas parenchyma echogenicity) - (tumor diameter < 10 mm) + 3. After receiver operating characteristic analysis, the area under the curve was 0.85. For score values of 5 (4) or more, sensitivity was 0.63 (0.93), specificity 0.91 (0.55), positive predictive value 0.82 (0.57), negative predictive value 0.79 (0.92), positive likelihood ratio 7.24 (2.05), and negative likelihood ratio 0.41 (0.14). CONCLUSION: A simple and potentially useful score for the prediction of pancreatic cancer based on six endosonographic criteria and patient age was established. Distribution of final diagnoses in the patient population argues for the score's applicability in clinical practice of a tertiary referral center and warrants prospective validation. PMID- 17427114 TI - Surgery in Fasciola hepatica pancreatitis: report of a case and review of literature. AB - Fasciola hepatica is a trematode rarely causing disease in humans. In symptomatic cases, while various pathologies such as damage to liver parenchyma, acute cholecystitis, and obstructive jaundice can be seen, the development of pancreatitis is rarely mentioned in the literature. The treatment of the disease is medical. In cases where no definite diagnosis can be made or in incidental cases where common bile duct exploration is being done, F. hepatica can be detected accidentally during operation. No consensus has yet been reached on the surgical procedure to be applied in this condition. We report on our case due to the rare occurrence of pancreatitis as a complication. In surgical cases, external drainage of the bile is both crucial in observing the response to the treatment, and also should be accepted as part of the treatment. PMID- 17427115 TI - Metal clip as a nidus for formation of common bile duct stone following laparascopic cholecystectomy. AB - A 83-year-old woman came to our hospital with fever up to 38.5 degrees C, abdominal pain and jaundice. She had undergone a cholecystectomy for symptomatic gallstones in 1992. The endoscopic retrograde cholangiography showed a dilated common bile duct with a free floating surgical clip with stone formation around this foreign body. After endoscopic sphincterotomy the extraction of the foreign body was successful. We found a two cm cholesterol stone the nidus of which was built by a Filshie-clip. The post endoscopic course was uneventful. PMID- 17427116 TI - [The abdominal compartment syndrome - widely unknown in gastroenterology]. AB - A sustained increase in intra-abdominal pressure may derange cardiovascular haemodynamics, respiratory and renal functions and finally lead to multi-organ failure. It is primarily seen in surgical intensive care units and is most frequently associated with small and large bowel surgery, vascular surgery, and abdominal trauma. An expert panel has defined elevated intra-abdominal pressure > 20 mmHg in conjunction with newly occurring organ dysfunction as "abdominal compartment syndrome" (ACS). This entity is not well recognised in gastroenterology, although ACS may occur as a complication of endoscopic perforation resulting in tension pneumoperitoneum. With the propagation of laparoscopic procedures it may be appropriate to emphasise the importance of intra-abdominal pressure monitoring in order to avoid this potentially lethal complication. PMID- 17427117 TI - [Surveillance colonoscopy in ulcerative colitis patients in Germany]. AB - It has been assumed that cancer surveillance colonoscopy in patients with ulcerative colitis is not conducted according to the guidelines in Germany. An inquiry of the self-help organisation German Crohn's Disease/Ulcerative Colitis Association (DCCV) among organisation members belonging to colorectal cancer risk groups confirmed that the number of biopsies taken during colonoscopy is less than that proposed by the guidelines. Only with 9.2 % of the risk group did a guideline-conformal colonoscopy take place. In more than 50 % of the cases less than 10 biopsies were taken. PMID- 17427118 TI - Clade age and not diversification rate explains species richness among animal taxa. AB - Animal taxa show remarkable variability in species richness across phylogenetic groups. Most explanations for this disparity postulate that taxa with more species have phenotypes or ecologies that cause higher diversification rates (i.e., higher speciation rates or lower extinction rates). Here we show that clade longevity, and not diversification rate, has primarily shaped patterns of species richness across major animal clades: more diverse taxa are older and thus have had more time to accumulate species. Diversification rates calculated from 163 species-level molecular phylogenies were highly consistent within and among three major animal phyla (Arthropoda, Chordata, Mollusca) and did not correlate with species richness. Clades with higher estimated diversification rates were younger, but species numbers increased with increasing clade age. A fossil-based data set also revealed a strong, positive relationship between total extant species richness and crown group age across the orders of insects and vertebrates. These findings do not negate the importance of ecology or phenotype in influencing diversification rates, but they do show that clade longevity is the dominant signal in major animal biodiversity patterns. Thus, some key innovations may have acted through fostering clade longevity and not by heightening diversification rate. PMID- 17427119 TI - Adaptive advantages of cooperative courtship for subordinate male lance-tailed manakins. AB - Male lance-tailed manakins (Chiroxiphia lanceolata) cooperate in complex courtship displays, but the dominant (alpha) partner monopolizes mating opportunities. This raises the question of why subordinates (betas) cooperate. Three nonexclusive hypotheses explain the adaptive basis of helping behavior by subordinate males: cooperation may increase (1) subordinates' immediate reproductive success, (2) the reproductive success of close relatives, or (3) subordinates' chances of future reproduction. I demonstrated that beta males rarely sired chicks and were unrelated to their alpha partners but received delayed direct benefits from cooperation; betas had an increased probability of becoming an alpha when compared to males that had not been betas. To investigate the mechanism by which betas attain these adaptive benefits, I examined betas' success in replacing their alpha partners both in natural turnover events and when alphas were experimentally removed. Beta males did not consistently inherit alpha roles in the same territories where they served their beta tenure, arguing that queuing for status does not fully explain the benefits of cooperation for betas. Instead, betas may be apprenticing to develop effective and appropriate displays that enhance their subsequent success as alphas. Complex social affiliations appear to mediate selective pressure for cooperation in this species. PMID- 17427120 TI - Are species adapted to their regeneration niche, adult niche, or both? AB - Functional traits are important drivers of successional processes and the assembly of plant communities. It is generally assumed that functional traits are closely linked to the regeneration niche because of the high selection pressures in the seedling stage, but recent studies have challenged this view. In this study, I use cross species and phylogenetic correlation analysis between leaf traits and light environment to evaluate whether species are adapted to the regeneration niche, adult niche, or both. Leaf chemistry, morphology, physiology, and crown exposure were quantified for up to 58 Bolivian tropical moist forest tree species that differ in their regeneration and adult light niche. Multiple regression analysis shows that leaf traits of seedlings, saplings, and trees are most strongly related to the regeneration niche, and once this is taken into account, adult niche does not significantly explain any of the remaining variation in leaf traits. This suggests that, although the regeneration phase is short, it has a long-lasting effect on the form and shape of plant species. PMID- 17427121 TI - Adaptive trade-off in floral morphology mediates specialization for flowers pollinated by bats and hummingbirds. AB - Evolution toward increased specificity in pollination systems is thought to have played a central role in the diversification of angiosperms. Theory predicts that the presence of trade-offs in adapting to different pollinator types will favor specialization, yet few studies have attempted to characterize such interactions in nature. I conducted flight cage experiments with bats, hummingbirds, and artificial flowers to examine effects of corolla width on pollination. I videotaped visits to analyze pollinator behavior and counted pollen grains transferred to stigmas. Results demonstrated that flower-pollinator fit is critical to effective pollination; wide corollas guided bat snouts better, and narrow corollas guided hummingbird bills better. Poor fit resulted in variable entry angles and decreased pollen transfer. A model using these results predicts that wide corollas will be selected for when bats make more than 44% of the visits and narrow corollas when they make fewer. Intermediate corollas are never favored (i.e., generalization is always suboptimal). This is the first study to clearly document a pollinator-mediated fitness trade-off in floral morphology. PMID- 17427122 TI - Spiteful soldiers and sex ratio conflict in polyembryonic parasitoid wasps. AB - The existence of spiteful behaviors remains controversial. Spiteful behaviors are those that are harmful to both the actor and the recipient, and they represent one of the four fundamental types of social behavior (alongside selfishness, altruism, and mutual benefit). It has generally been assumed that the conditions required for spite to evolve are too restrictive, and so spite is unlikely to be important. This idea has been challenged in recent years, with the realization that localized competition can relax the conditions required for spite to evolve. Here we develop a theoretical model for a prime candidate for a spiteful behavior, the production of the sterile soldier caste in polyembryonic wasps. Our results show that (a) the biology of these soldiers is consistent with their main role being to mediate conflict over the sex ratio and not to defend against competitors and (b) greater conflict will occur in more outbred populations. We also show that the production of the sterile soldier caste can be classed as a spiteful behavior but that, to an extent, this is merely a semantic choice, and other interpretations such as altruism or indirect altruism are valid. However, the spite interpretation is useful in that it can lead to a more natural interpretation of relatedness and facilitate the classification of behaviors in a way that emphasizes biologically interesting differences that can be empirically tested. PMID- 17427123 TI - Adaptable night camouflage by cuttlefish. AB - Cephalopods are well known for their diverse, quick-changing camouflage in a wide range of shallow habitats worldwide. However, there is no documentation that cephalopods use their diverse camouflage repertoire at night. We used a remotely operated vehicle equipped with a video camera and a red light to conduct 16 transects on the communal spawning grounds of the giant Australian cuttlefish Sepia apama situated on a temperate rock reef in southern Australia. Cuttlefish ceased sexual signaling and reproductive behavior at dusk and then settled to the bottom and quickly adapted their body patterns to produce camouflage that was tailored to different backgrounds. During the day, only 3% of cuttlefish were camouflaged on the spawning ground, but at night 86% (71 of 83 cuttlefish) were camouflaged in variations of three body pattern types: uniform (n=5), mottled (n=33), or disruptive (n=34) coloration. The implication is that nocturnal visual predators provide the selective pressure for rapid, changeable camouflage patterning tuned to different visual backgrounds at night. PMID- 17427124 TI - Communication of male quality in owl hoots. AB - The evolution of communication through intrasexual selection is expected to lead signalers to transmit honest information on their fighting ability. Here we studied the information encoded in the acoustic structure of the territorial calls of a nocturnal raptor. During territorial contests, male scops owls give hoots composed of a downward frequency shift followed by a stable plateau. We found that the frequency of the hoot was negatively correlated with the body weight of the vocalizer. We shifted the frequency contour of natural hoots in order to create resynthesized calls corresponding to individuals of varying body weight and used these stimuli in playback experiments simulating an intrusion into the territory of established breeders. Territory owners responded less intensely when they heard hoots simulating heavier intruders, and males with heavier apparent weight tended to give hoots with a lower plateau in response to playbacks simulating heavier intruders. PMID- 17427125 TI - The shapes of phylogenetic trees of clades, faunas, and local assemblages: exploring spatial pattern in differential diversification. AB - Life on Earth is characterized by strong diversity skewness: related lineages typically show pronounced variation in diversification success, and clades contain hyperdiverse and depauperate subclades. Previous studies have documented diversity skewness only for entire (global) clades. We demonstrate methods for measurement and significance testing of diversity skewness of local assemblages and regional biotas; we illustrate this with an analysis of geographic structure in diversity skewness of primate assemblages. For primates, continental faunas differ in diversity skewness from expectations based on the global phylogeny: South American faunas have significantly low skewness and African faunas have significantly high skewness. However, no local assemblage has diversity skewness different from that expected based on sampling the continental fauna. We also document a latitudinal gradient in diversity skewness for the African assemblages and test for (but do not find) associations of skewness with longitude, local species richness, and net primary productivity. Our data suggest that continental scale biogeographic events rather than local-scale processes have shaped diversity skewness in modern primate faunas. PMID- 17427126 TI - Plastic trait integration across a CO2 gradient in Arabidopsis thaliana. AB - Shifts across environments in patterns of trait integration may govern or alter adaptive responses. Changes in resource supply rates may be an especially important cause of plasticity of trait integration because they can lead to shifts in colimitation and coregulation of traits. Traditional evolutionary genetic characterization of trait integration relies on covariance analyses. Structural equation modeling (SEM) can complement such analyses. The SEM provides insights into causal structure not possible with a covariance analysis, thereby providing mechanistic understanding of shifts in integration and suggesting likely foci of selection in changing environments. We tested for changes in trait integration by growing 35 genotypes of Arabidopsis thaliana (Brassicaceae; mouse eared cress) from throughout the species' range in four atmospheric CO2 concentrations: 250 (past), 355 (approximately recent CO2), 530, and 710 (future) microM M(-1). SEM revealed significant shifts in the integration of N, C, and H2O use and their effects on reproductive dry mass across the CO2 gradient. The low CO2 stress of 250 microM M(-1) had the most divergent integration structures. Standardized total effects of C assimilation, water loss, and early N mass changed in sign across the C supply gradient, and the total effect of quantum yield decreased from significant to nonsignificant values across the gradient. Transpiration exhibited significant genetic variation and is thus a candidate target for selection and adaptation under novel growth CO2 concentrations. The strength of the correlation between C assimilation and transpiration declined by 19% from 250 to 710 microM M(-1), indicating a partial decoupling of their current mutual evolutionary constraint in the atmosphere of the future. Structural equation analysis of functional integration provides unique insights into the mechanisms through which changes in limiting resources can alter the nature of trait integration. PMID- 17427127 TI - Fitness effects associated with the major flowering time gene FRIGIDA in Arabidopsis thaliana in the field. AB - To date, the effect of natural selection on candidate genes underlying complex traits has rarely been studied experimentally, especially under ecologically realistic conditions. Here we report that the effect of selection on the flowering time gene FRIGIDA (FRI) reverses depending on the season of germination and allelic variation at the interacting gene FLOWERING LOCUS C (FLC). In field studies of 136 European accessions of Arabidopsis thaliana, accessions with putatively functional FRI alleles had higher winter survival in one FLC background in a fall-germinating cohort, but accessions with deletion null FRI alleles had greater seed production in the other FLC background in a spring germinating cohort. Consistent with FRI's role in flowering, selection analyses suggest that the difference in winter survival can be attributed to time to bolting. However, in the spring cohort, the fitness difference was associated with rosette size. Our analyses also reveal that controlling for population structure with estimates of inferred ancestry and a geographical restriction was essential for detecting fitness associations. Overall, our results suggest that the combined effects of seasonally varying selection and epistasis could explain the maintenance of variation at FRI and, more generally, may be important in the evolution of genes underlying complex traits. PMID- 17427128 TI - Selection through female fitness helps to explain the maintenance of male flowers. AB - Andromonoecy, the production of both male and hermaphrodite flowers in the same individual, is a widespread phenomenon that occurs in approximately 4,000 species distributed in 33 families. Hypotheses for the evolution of andromonoecy suggest that the production of intermediate proportions of staminate flowers may be favored by selection acting through female components of fitness. Here we used the andromonoecious herb Solanum carolinense to determine the pattern of selection on the production of staminate flowers. A multivariate analysis of selection indicates that selection through female fitness favors the production of staminate flowers in at least one population. We conclude that this counterintuitive benefit of staminate flowers on female fitness highlights the importance of considering female components of fitness in the evolution of andromonoecy, a reproductive system usually interpreted as a "male" strategy. PMID- 17427129 TI - Scaling of nectar foraging in orchid bees. AB - Morphology influences the rate at which foraging bees visit nectar flowers, the quantity of nectar they must consume to fuel their activities, and, consequently, the profitability of flower species. Because feeding time is a major determinant of visitation rate, I used a biomechanical model to examine how energy intake rate (E) varies with sucrose concentration, body mass (M), and proboscis length in orchid bees (Apidae: Euglossini). Under geometric scaling, the optimal sugar concentration (Smax) should be largely independent of body size, and E proportional to M1.0. In a comparative study of 30 orchid bee species ranging from 50 to 800 mg, Smax fell between 35% and 40% w/w, but E proportional to M0.54, significantly less than model predictions. Proboscis length and radius scale geometrically with body mass, but proboscis length exhibits substantial size-independent variation, particularly in small bees. One cost of a long proboscis is a reduction in both E and Smax in accordance with the scaling model. This finding highlights a difference between the lapping mechanism used by bumblebees and the suction mechanism used by orchid bees. A field study confirms that orchid bees harvest nectars with between 34% and 42% sucrose, independent of body size. PMID- 17427130 TI - Habitat choice in predator-prey systems: spatial instability due to interacting adaptive movements. AB - The role of habitat choice behavior in the dynamics of predator-prey systems is explored using simple mathematical models. The models assume a three-species food chain in which each population is distributed across two or more habitats. The predator and prey adjust their locations dynamically to maximize individual per capita growth, while the prey's resource has a low rate of random movement. The two consumer species have Type II functional responses. For many parameter sets, the populations cycle, with predator and prey "chasing" each other back and forth between habitats. The cycles are driven by the aggregation of prey, which is advantageous because the predator's saturating functional response induces a short-term positive density dependence in prey fitness. The advantage of aggregation in a patch is only temporary because resources are depleted and predators move to or reproduce faster in the habitat with the largest number of prey, perpetuating the cycle. Such spatial cycling can stabilize population densities and qualitatively change the responses of population densities to environmental perturbations. These models show that the coupled processes of moving to habitats with higher fitness in predator and prey may often fail to produce ideal free distributions across habitats. PMID- 17427131 TI - Direct and indirect effects of a sex-biased antagonist on male and female fertility: consequences for reproductive trait evolution in a gender-dimorphic plant. AB - Gender-dimorphic plants are often subject to sex-differential enemy attack, but whether and how this contributes to trait evolution is unknown. To address this gap, we documented the spatiotemporal prevalence of sex-biased weevil damage in a gynodioecious strawberry. We then conducted path analysis to evaluate the direct and indirect pathways for weevils to affect female and male fertility and to mediate selection in two experimental gardens. Direct effects of weevils significantly reduced fertility and mediated selection on reproductive traits, even in the nonpreferred sex (females). Weevils significantly reduced floral display size in hermaphrodites in both gardens, and this translated into a substantial negative indirect effect on male fertility in the garden where the pathway to fertility via display was stronger. Thus, indirect effects of weevils can contribute to selection in hermaphrodites, which gain the majority of their fitness via male function. Our results also indicate that weevils often play a larger role than pollinators in shaping reproductive phenotype and thus raise the intriguing possibility that antagonists may be drivers of sexual dimorphism. Finally, our results support the view that mutualists, antagonists, and the abiotic environment should be considered when attempting to understand reproductive trait evolution in gender-dimorphic species. PMID- 17427132 TI - Short-term foraging costs and long-term fueling rates in central-place foraging swans revealed by giving-up exploitation times. AB - Foragers tend to exploit patches to a lesser extent farther away from their central place. This has been interpreted as a response to increased risk of predation or increased metabolic costs of prey delivery. Here we show that migratory Bewick's swans (Cygnus columbianus bewickii), though not incurring greater predation risks farther out or delivering food to a central place, also feed for shorter periods at patches farther away from their roost. Predictions from an energy budget model suggest that increasing metabolic travel costs per se are responsible. Establishing the relation between intake rate and exploitation time enabled us to express giving-up exploitation times as quitting harvest rates (QHRs). This revealed that net QHRs were not different from observed long-term net intake rates, a sign that the birds were maximizing their long-term net intake rate. This study is unique because giving-up decisions were measured at the individual level, metabolic and predation costs were assessed simultaneously, the relation with harvest rate was made explicit, and finally, short-term giving up decisions were related to long-term net intake rates. We discuss and conceptualize the implications of metabolic traveling costs for carrying-capacity predictions by bridging the gap between optimal-foraging theory and optimal migration theory. PMID- 17427133 TI - A macroevolutionary explanation for energy equivalence in the scaling of body size and population density. AB - Across a wide array of animal species, mean population densities decline with species body mass such that the rate of energy use of local populations is approximately independent of body size. This "energetic equivalence" is particularly evident when ecological population densities are plotted across several or more orders of magnitude in body mass and is supported by a considerable body of evidence. Nevertheless, interpretation of the data has remained controversial, largely because of the difficulty of explaining the origin and maintenance of such a size-abundance relationship in terms of purely ecological processes. Here I describe results of a simulation model suggesting that an extremely simple mechanism operating over evolutionary time can explain the major features of the empirical data. The model specifies only the size scaling of metabolism and a process where randomly chosen species evolve to take resource energy from other species. This process of energy exchange among particular species is distinct from a random walk of species abundances and creates a situation in which species populations using relatively low amounts of energy at any body size have an elevated extinction risk. Selective extinction of such species rapidly drives size-abundance allometry in faunas toward approximate energetic equivalence and maintains it there. PMID- 17427134 TI - Importance of interactions between food quality, quantity, and gut transit time on consumer feeding, growth, and trophic dynamics. AB - Ingestion kinetics of animals are controlled by both external food availability and feedback from the quantity of material already within the gut. The latter varies with gut transit time (GTT) and digestion of the food. Ingestion, assimilation efficiency, and thus, growth dynamics are not related in a simple fashion. For the first time, the important linkage between these processes and GTT is demonstrated; this is achieved using a biomass-based, mechanistic multinutrient model fitted to experimental data for zooplankton growth dynamics when presented with food items of varying quality (stoichiometric composition) or quantity. The results show that trophic transfer dynamics will vary greatly between the extremes of feeding on low-quantity/high-quality versus high quantity/low-quality food; these conditions are likely to occur in nature. Descriptions of consumer behavior that assume a constant relationship between the kinetics of grazing and growth irrespective of food quality and/or quantity, with little or no recognition of the combined importance of these factors on consumer behavior, may seriously misrepresent consumer activity in dynamic situations. PMID- 17427135 TI - Distinguishing microsite and competition processes in tree growth dynamics: an a priori spatial modeling approach. AB - Spatially oriented studies have examined the role of competition on plant populations and communities but not the combined effects of microsite heterogeneity and competition. The aim of this study was threefold: first, to apply and test a common geostatistical tool (semivariograms) to disentangle competition and microsite effects; second, to assess the results of this methodology against a generalized early stand development model for tree populations; and third, to examine the role and timing of microsite and competition processes in early population stages. We mapped and measured annual relative growth rates of trees in three different-aged ponderosa pine stands in Patagonia, Chile. We tested the relative support of five a priori semivariogram based hypotheses and showed that through stand development, many sites followed our expected sequence of semivariogram models. These translated to initial spatially random growth followed by microsite-dominated, mixed microsite and competition, and finally pure competition effects on growth. Our approach will have many and diverse applications wherever processes differ in the type of spatial pattern they exhibit as well as in spatial scale. We emphasize that this methodology works best when there is strong a priori support for the hypotheses being tested but the timing, strength, and occurrence of processes are not known. PMID- 17427136 TI - Can population genetic structure be predicted from life-history traits? AB - Population genetic structure is a key parameter in evolutionary biology. Earlier comparative studies have shown that genetic structure depends on species ecological attributes and life-history traits, but species phylogenetic relatedness had not been accounted for. Here we reevaluate the relationships between genetic structure and species traits in seed plants. Each species is characterized by a set of life-history and ecological features as well as by its geographic range size, its heterozygote deficit, and its genetic structure at nuclear and organelle markers to distinguish between pollen- and seed-mediated gene flow. We use both a conventional regression approach and a method that controls for phylogenetic relationships. Once phylogenetic conservatism and covariation among traits are taken into account, genetic structure is shown to be related with only a few synthetic traits, such as mating system for nuclear markers and seed dispersal mode or geographic range size for organelle markers. Along with other studies on invasiveness or rarity, our work illustrates the fact that predicting the fate of species across a broad taxonomic assemblage on the basis of simple traits is rarely possible, a testimony of the highly contingent nature of evolution. PMID- 17427137 TI - A perfect storm: the combined effects on population fluctuations of autocorrelated environmental noise, age structure, and density dependence. AB - While it is widely appreciated that climate can affect the population dynamics of various species, a mechanistic understanding of how climate interacts with life history traits to influence population fluctuations requires development. Here we build a general density-dependent age-structured model that accounts for differential responses in life-history traits to increasing population density. We show that as the temporal frequency of favorable environmental conditions increases, population fluctuations also increase provided that unfavorable environmental conditions still occur. As good years accumulate and the number of individuals in a population increases, successive life-history traits become vulnerable to density dependence once a return to unfavorable conditions prevails. The stronger this ratcheting of density dependence in life-history traits by autocorrelated climatic conditions, the larger the population fluctuations become. Highly fecund species, and those in which density dependence occurs in juvenile and adult vital rates at similar densities, are most sensitive to increases in the frequency of favorable conditions. Understanding the influence of global warming on temporal correlation in regional environmental conditions will be important in identifying those species liable to exhibit increased population fluctuations that could lead to their extinction. PMID- 17427138 TI - Potential for Ebola transmission between gorilla and chimpanzee social groups. AB - Over the past decade Ebola hemorrhagic fever has emerged repeatedly in Gabon and Congo, causing numerous human outbreaks and massive die-offs of gorillas and chimpanzees. Why Ebola has emerged so explosively remains poorly understood. Previous studies have tended to focus on exogenous factors such as habitat disturbance and climate change as drivers of Ebola emergence while downplaying the contribution of transmission between gorilla or chimpanzee social groups. Here we report recent observations on behaviors that pose a risk of transmission among gorilla groups and between gorillas and chimpanzees. These observations support a reassessment of ape-to-ape transmission as an amplifier of Ebola outbreaks. PMID- 17427139 TI - Predation can increase the prevalence of infectious disease. AB - Many host-pathogen interactions are embedded in a web of other interspecific interactions. Recent theoretical studies have suggested that reductions in predator abundance can indirectly lead to upsurges in infectious diseases harbored by prey populations. In this note, we use simple models to show that in some circumstances, predation can actually increase the equilibrial prevalence of infection in a host, where prevalence is defined as the fraction of host population that is infected. Our results show that there is no complete generalization possible about how shifts in predation pressure translate into shifts in infection levels, without some understanding of host population regulation and the role of acquired immunity. Our results further highlight the importance of understanding the dynamics of nonregulatory pathogens in reservoir host populations and the understudied effects of demographic costs incurred by individuals that survive infection and develop acquired immunity. PMID- 17427140 TI - Is there really insufficient support for Tilman's R* concept? A comment on Miller et al. AB - Miller et al. (2005), in the American Naturalist (165:439-448), critically reviewed the applicability of Tilman's resource-ratio hypothesis. One of their conclusions was that only eight experimental papers support the R* concept, while five do not. We are familiar with some of the latter studies, and we question this conclusion. Our evaluation shows that 12 of the 13 articles investigated by Miller et al. support R* prediction, while one article does not fit the experimental conditions for a proper test. Moreover, the microbial and aquatic literature contains many more competition experiments consistent with the R* prediction. We therefore conclude that there is strong experimental support for the R* concept, at least from studies with bacteria, phytoplankton, and zooplankton. PMID- 17427141 TI - Evaluating support for the resource-ratio hypothesis: a reply to Wilson et al. PMID- 17427142 TI - Oxygen, nitric oxide and articular cartilage. AB - Molecular oxygen is required for the production of nitric oxide (NO), a pro inflammatory mediator that is associated with osteoarthritis and rheumatoid arthritis. To date there has been little consideration of the role of oxygen tension in the regulation of nitric oxide production associated with arthritis. Oxygen tension may be particularly relevant to articular cartilage since it is avascular and therefore exists at a reduced oxygen tension. The superficial zone exists at approximately 6% O2, while the deep zone exists at less than 1% O2. Furthermore, oxygen tension can alter matrix synthesis, and the material properties of articular cartilage in vitro. The increase in nitric oxide associated with arthritis can be caused by pro-inflammatory cytokines and mechanical stress. Oxygen tension significantly alters endogenous NO production in articular cartilage, as well as the stimulation of NO in response to both mechanical loading and pro-inflammatory cytokines. Mechanical loading and pro inflammatory cytokines also increase the production of prostaglandin E2 (PGE2). There is a complex interaction between NO and PGE2, and oxygen tension can alter this interaction. These findings suggest that the relatively low levels of oxygen within the joint may have significant influences on the metabolic activity, and inflammatory response of cartilage as compared to ambient levels. A better understanding of the role of oxygen in the production of inflammatory mediators in response to mechanical loading, or pro-inflammatory cytokines, may aid in the development of strategies for therapeutic intervention in arthritis. PMID- 17427143 TI - [Gastroesophageal reflux disease (GERD) in patients with obstructive sleep apnoea syndrome (OSAS)]. AB - Gastroesophageal reflux disease (GERD) is a common cause of chronic cough, heartburn, epigastric or retrosternal discomfort, chest pain and abdominal pain or esophagitis. Our patients with OSAS seldom manifest GERD symptoms. We suspected that obesity and high pressure in abdominal cavity may induce acid gastroesophageal reflux in these patients. The aim of the study was to test the hypothesis that obesity, cigarettes smoking or ventilatory and gas exchange abnormalities provoke GERD. We studied 21 consecutive patients with severe OSAS (mean AHI 44.9+/-23.8) before CPAP treatment, all without GERD clinical symptoms. Standard polysomnography, gastroscopy and 24-h oesophageal pH monitoring was performed. There were 6 females, 15 males, mean age 57+/-9 years, mean BMI 38+/-6 kg/m2. All patients presented with normal spirometric and gas exchange values (mean VC 3.64+/-1.23 1, 90% of normal, mean FEV1 2.61+/-0.95 1, 83% of normal, mean FEV1%VC 72%, mean PaO2 68.1+/-7.7 mmHg, mean PaCO2 40.8+/-5.8 mmHg, mean pH 7.42+/-0.02). GERD was diagnosed in 14 patients. Patients with GERD were younger, more often were cigarettes smokers (5/14). We did not fi nd statistically significant differences between severity of OSAS, BMI, ventilatory or gas exchange parameters and GERD. PMID- 17427144 TI - [Safety of influenza vaccination in asthmatics]. AB - The common cause of exacerbations in asthmatics are viral infections. Influenza infection especially increases the risk of respiratory exacerbations. The ACIP report puts asthma among indications to regular influenza vaccinations. In practice, many physicians doubt in effectiveness and safety of vaccination in patients suffering from allergy. It caused such procedures were not included in asthma guidelines published by other authors. Our study confirmed the safety of influenza split vaccine vaccination in asthmatics. We proved that vaccination had not any influence on asthma control in 14-day period after injection. We showed also high frequency of, mostly local, side effects. These events were generally mild and transient. PMID- 17427145 TI - [Serum and sputum eosinophil cationic protein levels and clinical status in cystic fibrosis patients]. AB - Recent studies have emphasied the role of eosinophils and it's metabolites in the pathogenesis of lung disease in cystic fibrosis. This study was designed to assess the relationship between serum and sputum ECP levels and clinical status of cystic fibrosis patients. MATERIAL AND METHODS: 30 patients, aged 6-30 with moderate cystic fibrosis were recruited. Spirometry, weight and high, and Shwachman-Kulczycki score were measured, and serum and sputum samples were obtained for measurements of the eosinophil cationic protein (ECP). RESULTS: We observed significant inverse correlation between sputum ECP levels and BMI (p<0.001) and FEV1 (p<0.001), and not significant inverse correlation between sputum ECP levels Shwachman-Kulczycki score (p=0.057). There was no signifi cant correlation between sputum ECP levels and FEF25/75% and between serum ECP levels and measured clinical parameters. CONCLUSIONS: Results of this study suggest influence of eosinophil inflammation in respiratory tract on clinical status of CF patients. PMID- 17427146 TI - [Tobacco smoking among the first-year medical students]. AB - The studies have shown that despite studying medicine the rate of tobacco smokers among students is still high. Moreover, in Poland the incidence of smoking is increasing among the youngsters. The study was designed to assess the rate of smokers among the first-year students of two consecutive courses at the Medical University of Gdansk and define their attitudes towards tobacco smoking. The voluntary, multiple-choice questionnaires were distributed among 412 students. The rate of response was 100%. Twenty one percent of responders were current smokers (17% females and 28% men), whereas 7% declared previous smoking. There was no correlation between incidence of smoking among students and their parents (p=0.11). 61% of smokers declared the will to give up their habit and of those 94% were aware of the negative impact of cigarettes on their health. In the group which did not declare the will to quit smoking only 75% knew what the impact of cigarettes on their health was; p=0.02. Only 23% of smokers considered anti nicotine therapy effective. Almost a half of smokers considered nicotine replacement therapy ineffective for them in quitting cigarette smoking. CONCLUSION: The rate of smokers among the first-year medical students is lower than in general population, but it is still relatively high. However more than a half of smokers wants to give up their habit. Of those who are not willing to, 25% is not aware of the cigarettes' impact on their health. These results call for further anti-nicotine actions. PMID- 17427147 TI - [Respiratory responses to CO2 stimulation in hypercapnic patients with obstructive sleep apnea syndrome]. AB - Obstructive sleep apnea can be associated with daytime chronic hypercapnia in some patients, but the prevalence of the phenomenon is highly variable in the published literature. The most often it is found in patients with coexisting COPD. There is also an evidence of persisting hypercapnia in OSA patients without other respiratory disease. In previous studies lung function impairment, obesity, gender, severity of OSAS have been considered to contribute to daytime hypercapnia. Several studies demonstrated that the defect in control of breathing can play a role in the development of chronic hypercapnia in patients with OSAS. The aim of the study was to estimate respiratory responses to hypercapnic stimulation in patients with OSAS and chronic daytime hypercapnia. Material consisted of 38 patients with OSAS and chronic hypercapnia (COPD was present in 24-group B, "pure" OSA in 14-group A) and 40 normocapnic OSA patients (group C). Lung function testing, blood gases and chemical control of breathing tests were performed in all of them before initiating therapy with nCPAP. Diagnosis of OSAS was stated with standard polisomnography and AHI was similar in mentioned groups. RESULTS: Respiratory responses to hypercapnic stimulation were significantly lower in hypercapnic patients (A 10.6+/-4.6; B 9.5+/-5.6) in opposition to normocapnics (C 23.3+/-14.0 l/min/kPa). In all studied patients PaCO2 level significantly correlated with respiratory responses to hypercapnic stimulation (r=-.61), lung function indices (VC r=-.69 and FEV1 r=-.71), mean SaO2 during sleep (r=-.68), and BMI (r=.49), but not with the factors like age, AHI or minimal SaO2 during sleep. Analysis with multiple regression revealed that hypercapnic drive, mean SaO2 during sleep, FEV1 and BMI were the best predictors of hypercapnia in studied group, being responsible for 72% of the total variance in PaCO2 in our OSA patients (R2=0.72; p<0.0001). CONCLUSION: predisposition to daytime hypercapnia in our OSA patients was related to dimished chemosensitivity to CO2, mean desaturation during sleep, the severity of obesity and impairment of lung function mainly due to coexisting COPD. PMID- 17427148 TI - [Lack of correlation between exhaled nitric oxide (eNO) and clinical indicators of the disease activity and quality of life in mild and moderate asthmatics]. AB - The measurements of exhaled nitric oxide (eNO) are simple and useful method of assessment of inflammation in asthmatics' airways. One of the causes of its limited application in clinical practice is a number of factors influencing the results of measurements. The aim of the study was to determine the usefulness of eNO measurements in assessing the inflammation in a heterogeneous, in relation to atopic and smoking status, group of patients. MATERIALS AND METHODS: 120 subjects suspected of having asthma participated in this study. During 2 weeks the patients noted daily asthma symptoms and daily use of rescue medication. After 14 days health related quality of life (HRQL) was determined by means of Asthma Quality of Life Questionnaire (AQLQ), eNO levels were measured and airways reversibility test was performed. RESULTS: Preliminary diagnosis of asthma was confirmed in 84 patients on the basis of positive result of airways reversibility test. Among them, 21 subjects (25%) were smokers and 60 (71.4%) were atopic. No correlation was found between eNO and daily asthma symptom score, daily use of rescue medication, percent of airway reversibility after beta2-agonist and HRQL. CONCLUSION: eNO measurements in a heterogeneous, in relation to atopic and smoking status, group of patients are of limited value in clinical assessment of asthma activity. PMID- 17427149 TI - [Exacerbations of chronic obstructive pulmonary disease and the role of sputum bacteriological examination]. AB - Bacteriological examination of sputum is the simplest and widely accessible diagnostic method of respiratory infections. However its value in nonspecific respiratory infections, especially in exacerbations of COPD, is questionable because they can be caused by factors other than bacterial or by viral infections. The evaluation of bacteriological examination of sputum in patients with exacerbations of COPD and the evaluation of interaction between clinical course, some laboratory markers and bacteriology of sputum was the aim of the study. 109 patients hospitalized with exacerbations of COPD were examined. Semi quantitative bacteriological examination of sputum, total blood count, erythrocytes sedimentation rate, gasometry and spirometry were performed in each patient. The identification of pathogens was conducted by microtests API from Bio Merieux. In 39 patients (36%) pathogenic bacteria were cultured from sputum. The most prevalent organisms were: A. baumanii-21% and S. aureus-17%. Positive culture was seen most often in patients with severe and very severe COPD. Bacterial infection as a cause of COPD exacerbation should be suspected especially in patients with severe-staged disease of long duration, when bacterial cells and predominant neutrophil-count are present in sputum. In patients with severe COPD, often treated in hospital and with antibiotics, Gram negative flora should be considered as an etiologic agent. PMID- 17427150 TI - [Exacerbation of idiopathic pulmonary fibrosis]. PMID- 17427151 TI - [The role of interleukin 17 cytokine family in inducing allergic inflammation in the pulmonary tract]. AB - IL-17 family is a group of proinflammatory cytokines produced by activated memory T-cells. These cytokines play an important role in the development of cellular and humoral mechanisms of immunological responses lying at the basis of allergic disorders. The aim of this paper is to present the current knowledge on the role of interleukin 17 cytokine family in the pathogenesis of allergic disorders of the respiratory tract. IL-17A (as well as IL-17F) plays role in the development of airway hyperresponsiveness through activation of allergen-specific T-cells. Levels of IL-17A are elevated in sputum of asthmatic patients and correlate with airway hyperresponsiveness to methacholine. However, it remains fact, that the main effect of IL-17A in the pulmonary tract is recrutation of polymorphonuclear leukocytes, depending on CXC chemokine release from stromal cells. IL-17E evokes different immunological responses. This cytokine participates in the development of Th2-cell-dependent immunological response and the coexisting pathological tissue changes. These actions take place mainly through the induction of synthesis of the Th2 cell-derived cytokines (IL-4, IL-5, IL-13) and the development of eosinophilic inflammation. It is thought, that the character of the immunological response evoked by different cytokines of IL-17 family depends on the differences between the spatial structure of their fragments including disulfide bridges and that these differences determine their receptor interactions and biological functions. PMID- 17427152 TI - [Angiogenesis and neoangiogenesis--the role in lung cancer and other tumors]. PMID- 17427153 TI - [Cardiac involvement in sarcoidosis]. PMID- 17427154 TI - [Central sleep apnoea (CSA) in male with heart failure]. AB - We studied 44-year old man with heart failure (ejection fraction -25%). Obesity, arterial hypertension, snoring and excessive daytime sleepiness suggested concomitant obstructive sleep apnoea. Limited polysomnography with Polymesam revealed typical Cheyne-Stokes respiration with mainly central apnoeas (RDI=48/hour). We did not find any obstructive episodes during sleep study. Patient responded to CPAP therapy and apnoea hypopnoe index decreased to 12/hour on 8 mbar pressure. PMID- 17427155 TI - [History pages]. PMID- 17427156 TI - [Dr Mateusz Fonberg (1840-1880)]. PMID- 17427157 TI - [Miraculous survival]. PMID- 17427160 TI - pcaH, a molecular marker for estimating the diversity of the protocatechuate degrading bacterial community in the soil environment. AB - Microorganisms degrading phenolic compounds play an important role in soil carbon cycling as well as in pesticide degradation. The pcaH gene encoding a key ring cleaving enzyme of the beta-ketoadipate pathway was selected as a functional marker. Using a degenerate primer pair, pcaH fragments were cloned from two agricultural soils. Restriction fragment length polymorphism (RFLP) screening of 150 pcaH clones yielded 68 RFLP families. Comparison of 86 deduced amino acid sequences displayed 70% identity to known PcaH sequences. Phylogenetic analysis results in two major groups mainly related to PcaH sequences from Actinobacteria and Proteobacteria phyla. This confirms that the developed primer pair targets a wide diversity of pcaH sequences, thereby constituting a suitable molecular marker to estimate the response of the pca community to agricultural practices. PMID- 17427161 TI - Cell cycle effects resulting from inhibition of hepatocyte growth factor and its receptor c-Met in regenerating rat livers by RNA interference. AB - Hepatocyte growth factor (HGF) and its receptor c-Met are involved in liver regeneration. The role of HGF and c-Met in liver regeneration in rat following two-thirds partial hepatectomy (PHx) was investigated using RNA interference to silence HGF and c-Met in separate experiments. A mixture of 2 c-Met-specific short hairpin RNA (ShRNA) sequences, ShM1 and ShM2, and 3 HGF-specific ShRNA, ShH1, ShH3, and ShH4, were complexed with linear polyethylenimine. Rats were injected with the ShRNA/PEI complex 24 hours before and at the time of PHx. A mismatch and a scrambled ShRNA served as negative controls. ShRNA treatment resulted in suppression of c-Met and HGF mRNA and protein compared with that in controls. The regenerative response was assessed by PCNA, mitotic index, and BrdU labeling. Treatment with the ShHGF mixture resulted in moderate suppression of hepatocyte proliferation. Immunohistochemical analysis revealed severe suppression of incorporation of BrdU and complete absence of mitosis in rats treated with ShMet 24 hours after PHx compared with that in controls. Gene array analyses indicated abnormal expression patterns in many cell-cycle- and apoptosis related genes. The active form of caspase 3 was seen to increase in ShMet-treated rats. The TUNEL assay indicated a slight increase in apoptosis in ShMet-treated rats compared with that in controls. CONCLUSION: The data indicated that in vivo silencing of c-Met and HGF mRNA by RNA interference in normal rats results in suppression of mRNA and protein, which had a measurable effect on proliferation kinetics associated with liver regeneration. PMID- 17427162 TI - Daytime sources of nitrous acid (HONO) in the atmospheric boundary layer. AB - Nitrous acid (HONO) is an important precursor of the hydroxyl radical (OH), the self-cleaning agent of the atmosphere and a key species in the formation of harmful photooxidants during summer smog. Recent field measurements using very sensitive HONO instruments have shown that daytime HONO concentrations are much higher than has been assumed previously and that the contribution of HONO to the radical formation was underestimated in the past. A strong photochemical HONO source has been proposed, which contributes to the primary OH radical production up to 56 %. These exciting results initiated new laboratory studies, in which new sources of HONO have been identified. It is demonstrated that HONO is photochemically formed 1) on surfaces treated with nitric acid, 2) by reduction of NO(2) on photosensitized organic surfaces like humic acids and c) in the gas phase photolysis of ortho-substituted nitroaromatics. Although significant uncertainties still exist on the exact mechanisms, these additional sources might explain daytime observations in the atmosphere and demonstrate that HONO should be generally measured in field campaigns, besides other radical sources. PMID- 17427163 TI - Magnetic properties and crystal structure of solvent-free Sc@C82 metallofullerene microcrystals. AB - Magnetic properties of solvent-free crystals of the endohedral Sc@C82 are investigated by SQUID and X-ray powder diffraction. We find that the crystal is a paramagnet and the magnetic susceptibility decreases from 150 K with evidence of antiferromagnetic-like interactions by slow cooling. X-ray crystal analysis reveals the presence of a phase transition at 150 K, which is attributed to an orientational ordering transition of the fullerene molecules. At low temperatures we find a magnetic metastable state that can be controlled by the cooling rate. The metastable state can be formed by rapid cooling. The direction of Sc@C82 molecular axis in the crystals is disordered in the metastable state, and the susceptibility is higher than that in the slow cooling case at low temperature. PMID- 17427164 TI - An alkoxide-directed intermolecular [2+2+1] annulation: a three-component coupling reaction for the synthesis of tetrasubstituted alpha,beta-unsaturated gamma-lactams. PMID- 17427165 TI - Cu2(OTf)2-catalyzed and microwave-controlled preparation of tetrazoles from nitriles and organic azides under mild, safe conditions. PMID- 17427166 TI - Ring expansion of a Cp* moiety: formation of a 1,2-diphosphacyclooctatetraene ligand. PMID- 17427167 TI - The aggregated state of amyloid-beta peptide in vitro depends on Cu2+ ion concentration. PMID- 17427168 TI - Targeted RNA interference of phosphatidylinositol 3-kinase p110-beta induces apoptosis and proliferation arrest in endometrial carcinoma cells. AB - Phosphatidylinositol 3-kinase (PI3K) signalling plays a pivotal role in intracellular signal transduction pathways involved in cell growth, cellular transformation, and tumourigenesis. PI3K is overexpressed in many human cancers, including endometrial carcinomas, one of the most common female genital tract malignancies. Here, we used small interfering RNA (siRNA) targeted to PI3K p110 beta to determine whether inhibition of the beta isoform could be a potential therapeutic target for endometrial carcinoma. In this study, treatment of HEC-1B endometrial cancer cells with PI3K p110-beta-specific siRNA resulted in increased apoptosis and decreased tumour cell proliferation. Depletion of PI3K p110-beta decreased the protein levels of AKT1, AKT2, pAKT, and mTOR-downstream targets of PI3K. Knock-down of PI3K p110-beta by siRNA also induced decreased expression of cyclin E and Bcl-2, suggesting that PI3K p110-beta stimulates tumour growth, at least in part by regulating cyclin E and Bcl-2. Thus, our results indicate that siRNA-mediated gene silencing of PI3K p110-beta may be a useful therapeutic strategy for endometrial cancers overexpressing PI3K p110-beta. PMID- 17427169 TI - Role of BRAF-V600E in the serrated pathway of colorectal tumourigenesis. AB - There is increasing evidence for an alternative pathway of sporadic colorectal tumourigenesis that is associated with DNA microsatellite instability (MSI), due to methylation and loss of expression of the mismatch repair gene MLH1. Recent studies have highlighted a serrated pathway of colorectal cancer (CRC) in which serrated polyps with activating mutations in BRAF progress to CRCs with MSI following methylation and silencing of MLH1. The present study provides a novel mechanistic experimental model for these clinical observations. We investigated the role of BRAF activating mutation (BRAF-V600E) in colorectal tumourigenesis by studying the effects of forced expression of BRAF-V600E in the 'normal' colon epithelial NCM460 cell line and by targeting endogenous BRAF-V600E in MSI-High (MSI-H) colon cancer cell lines. The findings indicate that BRAF mutation in colon epithelial cells contributes to a gain in resistance towards apoptotic stimuli, which is likely to be an important characteristic of pre-malignant serrated lesions. BRAF-V600E also plays a role in the development and maintenance of transformed and invasive phenotypes in colon epithelial cells. Our findings also suggest that BRAF mutation potentiates promoter hypermethylation of the MLH1 gene promoter. Together, these results highlight BRAF as a potential target for therapeutic intervention in sporadic MSI-H colorectal cancers. PMID- 17427170 TI - Ultrasound-guided transcutaneous Tru-Cut biopsy to diagnose laryngopharyngeal masses: a pilot study. AB - BACKGROUND: Patients with bulky laryngopharyngeal masses and a relative or absolute contraindication to general anesthesia present diagnostic difficulties. In the current study, the authors assessed the utility of transcutaneous ultrasound-guided Tru-Cut biopsy (USGTCB) under local anesthesia in such individuals. METHODS: The current report was a prospective, nonrandomized study. Patients meeting the inclusion criteria underwent USGTCB as outpatients. RESULTS: Ten patients were recruited, 4 of whom had an untreated mass obstructing the laryngeal lumen and 6 of whom were previously treated for laryngeal cancer. All tolerated the procedure well. The only adverse event noted was spontaneously resolving near-syncope in 1 patient. In 9 patients the biopsy was diagnostic: invasive squamous cell carcinoma in 7 patients and chondronecrosis in 2 patients. In the remaining patient, radiologically suspected cricoid chondrosarcoma was confirmed based on the surgical specimen. CONCLUSIONS: The results of this pilot study are encouraging. USGTCB of laryngopharyngeal masses was found to produce no local morbidity, was diagnostic in each of the 5 patients with suspected disease recurrence after radiotherapy, was feasible in the outpatient setting, and had high sensitivity and specificity. The procedure is particularly useful for patients contraindicated for general anesthesia or those with a risk of tracheotomy due to intubation difficulties. PMID- 17427172 TI - Comparison of two techniques of transarterial chemoembolization before liver transplantation for hepatocellular carcinoma: a case-control study. AB - Supraselective transarterial chemoembolization (STACE) more efficiently targets chemotherapy delivered via the feeding arterial branches of the tumor than does conventional transarterial chemoembolization (TACE). However, the hypothesis of its greater efficacy compared with the latter is subject to controversy. The aim of the present study was to compare STACE to conventional TACE in a controlled study of candidates for liver transplantation (LT) for hepatocellular carcinoma (HCC). Patients were matched for factors associated with HCC recurrence and survival. Sixty patients were included: 30 who were treated with STACE and 30 treated with conventional TACE. The 2 groups were similar in terms of matched criteria. In the overall population (uni- and multinodular HCC), there was no marked difference between the 2 groups in 5-year disease-free survival: 76.8% vs. 74.8%. In sensitivity analysis of patients considered to be the best candidates for TACE (uninodular HCC < or =5 cm), there was a trend toward significance between STACE and TACE in 5-year disease-free survival: 87% vs. 64% (P = 0.09). The only factor associated with complete tumor necrosis was STACE in the overall population (30.8% vs. 6.9%, P = 0.02), with a similar trend in the subgroup of patients with a single nodule (33.3% vs. 6.7%, P = 0.06), whereas the mean number of procedures was similar in the 2 groups (mean, 1.3 procedures; range 1-5 procedures; P = NS). STACE is more efficient at inducing complete tumor necrosis in the liver. This study observed trends toward improvement in the disease-free survival of patients with uninodular HCC < or =5 cm. Future studies focusing on such patients are warranted. PMID- 17427173 TI - Vascular complications after orthotopic liver transplantation after neoadjuvant therapy for hilar cholangiocarcinoma. AB - Liver transplantation after neoadjuvant chemoradiotherapy has emerged as an effective treatment for patients with localized, node-negative, unresectable hilar cholangiocarcinoma (CCA) or CCA arising in the setting of primary sclerosing cholangitis (PSC). However, concern has arisen regarding the potential for vascular complications due to high-dose neoadjuvant therapy before transplantation. We reviewed our experience with specific aims to determine the incidences of arterial, portal, and hepatic venous complications in patients transplanted for CCA compared with patients who undergo transplantation for other indications, and to describe patient outcome as a result of these vascular complications. We reviewed data for all patients who underwent liver transplantation for CCA between January 1993 and April 2006 and compared the incidences of vascular complications to whole organ and living donor recipient control groups. Sixty-eight patients underwent neoadjuvant therapy and subsequent liver transplantation. Arterial complications arose in 21%; portal venous complications arose in 22%; and overall, 40% developed vascular complications. Late hepatic artery complications occurred more often in living donor recipients transplanted for CCA compared with the living donor control group (P=0.047). Late portal vein complications occurred more often in both whole organ and living donor recipients transplanted for CCA compared with the control groups (P=0.01 and P=0.009). Hepatic venous complications were rare. Patient and graft survival were not different between CCA and control patients. Liver transplantation with neoadjuvant therapy is associated with far higher rates of late arterial and portal venous complications, but these complications do not adversely affect patient and graft survival. PMID- 17427174 TI - Early and late outcome of cardiac surgery in patients with liver cirrhosis. AB - Liver cirrhosis is a major risk factor in general surgery. Few studies have reported on the outcome of cardiac surgery in these patients. Herein we report our recent experience in this high-risk patient population according to the Child Turcotte-Pugh classification and Model for End-Stage Liver Disease (MELD) score. Between January 1998 and December 2004, 27 patients (mean age 58 +/- 10 yr, 20 male) with cirrhosis who underwent cardiac surgery were identified. Patients were in Child-Turcotte-Pugh class A (n = 10), B (n = 11), and C (n = 6) and mean MELD score was 14.2 +/- 4.2. Operative mortality was 26% (n = 7). Stratified mortality according to Child-Turcotte-Pugh class was 11%, 18%, and 67% for class A, B, and C, respectively. No mortality occurred in patients who had revascularization without the use of cardiopulmonary bypass (n = 5). The 1-yr survival was 80%, 45%, and 16% for Child-Turcotte-Pugh class A, B, and C, respectively (P = 0.02). Major postoperative complications occurred in 22%, 56%, and 100% for Child Turcotte-Pugh class A, B, and C, respectively. Child-Turcotte-Pugh classification was a better predictor of hospital mortality (P = 0.02) compared to MELD score (P = 0.065). In conclusion, our results suggest that cardiac surgery can be performed safely in patients with Child-Turcotte-Pugh class A and selected patients with class B. Operative mortality remains high in class C patients. Careful patient selection is critical in order to improve surgical outcome in patients with cirrhosis. PMID- 17427175 TI - Increase in the toxic effects of Tl+ on isolated rat liver mitochondria in the presence of nonactin. AB - The effects of Tl(+) ions on isolated rat liver mitochondria were studied in the presence of nonactin, a cyclic ionophore. Nonenergized rat liver mitochondria were increasingly swollen at an elevated concentration of Tl(+) in the 160 mOsm medium containing 0-150 mM sucrose and 0-75 mM TlNO(3) or 0-50 mM Tl acetate. On the contrary, mitochondria in experiments with nonactin were contracted in the medium with 5-25 mM Tl(+) and were swollen only in the medium with 50-75 mM TlNO(3) or 50 mM Tl acetate. State 4 respiration along with swelling of succinate energized mitochondria followed contraction after their deenergization was further enhanced at increasing concentration of Tl acetate in a medium containing nonactin. Regardless of the presence of nonactin, State 3 and 2,4-dinitrophenol (DNP)-stimulated respiration and the monoamine oxidase (MAO) activity were not affected in the medium with 0-25 mM Tl acetate and sucrose. DNP-stimulated respiration decreased and the MAO activity somewhat increased in the medium containing 50 mM Tl acetate and nonactin. Uptake of (86)Rb(+) by energized mitochondria in the presence of valinomycin was considerably decreased when Tl(+) and nonactin were simultaneously present in the medium. An increase of the toxic effect of Tl(+) on rat liver mitochondria in the presence of nonactin is accounted for by disruption of mitochondria due to their more extensive swelling and uncoupling of mitochondria, resulting in the stimulation of State 4 and depletion of their energy store. PMID- 17427176 TI - Unaltered hepatic oxidative phosphorylation and mitochondrial permeability transition in wistar rats treated with nimesulide: Relevance for nimesulide toxicity characterization. AB - Nonsteroidal anti-inflammatory drugs have been associated with hepatotoxicity in susceptible patients. One such example is nimesulide, a preferential cyclooxygenase 2-inhibitor, widely used for the treatment of inflammation and pain. It was suggested that nimesulide could exert its hepatotoxicity by altering hepatic mitochondrial function, which was demonstrated in vitro. The objective of this study was to verify whether liver mitochondria isolated from rats treated with doses of nimesulide well above therapeutic levels possessed decreased calcium tolerance and oxidative phosphorylation, which indicates in vivo nimesulide mitochondrial toxicity. Male and female rats received nimesulide or its vehicle twice daily, for 5 days, and were killed on the seventh day for the isolation of liver mitochondria. Mitochondrial respiration, transmembrane electric potential, and calcium tolerance were characterized in all experimental groups. Nimesulide had no effect on liver mitochondrial function. Indexes of mitochondrial integrity, calcium loading capacity, and oxidative phosphorylation efficiency were unchanged between liver mitochondria from treated and control animals. In the animals tested, no evidence of degraded mitochondrial function due to nimesulide administration could be found. The results corroborate the notion that despite recognized in vitro mitochondrial toxicity, nimesulide does not cause detectable mitochondrial dysfunction in Wistar rats, even when administered in much higher concentrations than those known to have anti inflammatory effects. PMID- 17427177 TI - A Mu-class glutathione S-transferase from gills of the marine shrimp Litopenaeus vannamei: purification and characterization. AB - Glutathione S-transferases (GSTs) are a family of detoxifying enzymes that catalyze the conjugation of glutathione (GSH) to electrophiles, thereby increasing the solubility of GSH and aiding its excretion from the cell. In this study, a glutatione S-transferase from the gills of the marine shrimp Litopenaeus vannamei was purified by affinity chromatography using a glutathione-agarose affinity column. GST was purified to homogeneity as judged by reducing SDS-PAGE and zymograms. This enzyme is a homodimer composed of approximately 25-kDa subunits and identified as a Mu-class GST based on its activity against 1-chloro 2,4-dinitrobenzene (CDNB) and internal peptide sequence. The specific activity of purified GST was 440.12 micromol/(min mg), and the K(m) values for CDNB and GSH are very similar (390 and 335 microM, respectively). The intersecting pattern of the initial velocities of this enzyme in the Lineweaver-Burke plot is consistent with a sequential steady-state kinetic mechanism. The high specific activity of shrimp GST may be related to a highly effective detoxification mechanism necessary in gills since they are exposed to the external and frequently contaminated environment. PMID- 17427178 TI - Effect of aroclor 1248 and two pure PCB congeners on phospholipase D activity in rat renal tubular cell cultures. AB - This paper elucidates the effect of different polychlorinated biphenyls (PCBs) on the phospholipase D (PLD) activity in soluble and particulate fractions of rat renal proximal tubular culture cells. Treatment with Aroclor 1248 (a commercial PCB mixture) caused a marked increase in the activity of PLD in intact renal tubular cells. The PLD activity was increased by Aroclor 1248 in the particulate fraction while the enzyme activity was unaffected in the soluble fraction. This work also shows that PCB 153 (2,2',4,4',5,5'-hexachlorobiphenyl, a di-ortho substituted nonplanar congener) can increase the activity of PLD only in the particulate fraction. The exposure of cell cultures to PCB 77 (3,3',4,4' tetrachlorobiphenyl, a non-ortho-substituted planar congener) does not alter PLD activity. These results suggest that PCB effects are structure dependent. Therefore, in order to clarify the molecular mechanism of activation of PLD by PCBs, the contents of immunoreactive PLD were examined by immunoblot analysis. Renal tubular cells expressed a PLD protein of 120 kDa corresponding with the PLD1 mammalian isoform in both the particulate and the soluble fraction. Aroclor 1248, PCB 153, and PCB 77 do not induce changes in the levels of PLD protein. These data indicate that PCBs, particularly nonplanar congeners, increase PLD activity. Moreover, these changes could not be demonstrated in the enzyme content in rat renal tubular cell cultures. PMID- 17427179 TI - Inhibition of fipronil and nonane metabolism in human liver microsomes and human cytochrome P450 isoforms by chlorpyrifos. AB - Previous studies have established that chlorpyrifos (CPS), fipronil, and nonane can all be metabolized by human liver microsomes (HLM) and a number of cytochrome P450 (CYP) isoforms. However, metabolic interactions between these three substrates have not been described. In this study the effect of either coincubation or preincubation of CPS with HLM or CYP isoforms with either fipronil or nonane as substrate was investigated. In both co- and preincubation experiments, CPS significantly inhibited the metabolism of fipronil or nonane by HLM although CPS inhibited the metabolism of fipronil more effectively than that of nonane. CPS significantly inhibited the metabolism of fipronil by CYP3A4 as well as the metabolism of nonane by CYP2B6. In both cases, preincubation with CPS caused greater inhibition than coincubation, suggesting that the inhibition is mechanism based. PMID- 17427180 TI - Nonstereogenic alpha-aminoisobutyryl-glycyl dipeptidyl unit nucleates type I' beta-turn in linear peptides in aqueous solution. AB - The use of alpha,alpha-disubstituted amino acids represents a valuable strategy to exercise conformational control in peptides. Incorporation of the nonstereogenic alpha-aminoisobutyryl-glycyl (Aib-Gly) dipeptidyl sequence into i+1 and i+2 positions of an acyclic peptide sequence, originally designed and investigated by Gellman and coworkers, [H-Arg-Tyr-Val-Glu-Val-Yyy-Xxx-Orn-Lys-Ile Leu-Gln-NH2] nucleates a stable [2:4] left-handed type I' beta-turn in water. NMR spectra show that this newly designed beta-hairpin does not aggregate in water up to a concentration of approximately 1 mM, and that its backbone conformation is superimposable on corresponding hairpins containing the DPro-Gly (literature) and Aib-DAla (this work) sequences. The Aib-Gly turn-inducer sequence eliminates complications because of cis-trans isomerization of Zzz-Pro bonds, and constitutes an attractive alternative to the proteogenic Asn-Gly and nonproteogenic DPro-Gly motifs previously suggested as turn-inducer sequences. These design principles could be exploited to prepare water-soluble beta-hairpin peptides with robust structures and novel function. PMID- 17427181 TI - Targeting protein-protein interactions: lessons from p53/MDM2. AB - The tremendous challenge of inhibiting therapeutically important protein-protein interactions has created the opportunity to extend traditional medicinal chemistry to a new class of targets and to explore nontraditional strategies. Here we review a widely studied system, the interaction between tumor suppressor p53 and its natural antagonist MDM2, for which both traditional and nontraditional approaches have been reported. This system has been a testing ground for novel proteomimetic scaffold-based strategies, i.e., for attempts to mimic the recognition surface displayed by a folded protein with unnatural oligomers. Retroinverso peptides, peptoids, terphenyls, beta-hairpins, p oligobenzamides, beta-peptides, and miniproteins have all been explored as inhibitors of the p53/MDM2 interaction, and we focus on these oligomer-based efforts. Traditional approaches have been successful as well, and we briefly review small molecule inhibitors along with other strategies for reactivation of the p53 pathway, for comparison with oligomer- based approaches. We close with comments on an emerging dichotomy among protein-protein interaction targets. PMID- 17427182 TI - Caffeine challenge in patients with panic disorder: baseline differences between those who panic and those who do not. AB - A proportion of patients with panic disorder (PD) display an increased sensitivity to the anxiogenic/panicogenic properties of caffeine. The aim of this study is to identify probable baseline differences between PD patients who panic and those who do not, after caffeine administration. In a randomized, double blind, cross-over experiment performed in two occasions 3-7 days apart, 200 and 400 mg of caffeine, respectively, were administered in a coffee form to 23 patients with PD with or without Agoraphobia. Evaluations included the State Trait Anxiety Inventory, the DSM-IV 'panic attack' symptoms (visual analogue scale form), the Symptom Checklist-90-Revised (SCL-90-R), as well as breath holding (BH) duration, heartbeat perception accuracy and heart rate. Only those patients who did not present a panic attack after both challenges ('no panic group', N=14, 66.7%), and those who presented a panic attack after at least one challenge ('panic group', n=7, 33.3%) were included in the analysis. The panickers, compared to the non-panickers, presented at baseline: significantly higher total score of the SCL-90-R; significantly higher scores on all the SCL-90 R clusters of symptoms, except that of 'paranoid ideation'; significantly lower BH duration. The present preliminary findings indicate that PD patients who panic after a 200 mg or a 400 mg caffeine challenge, compared to the PD patients who do not panic after both of these challenges, may present at baseline significantly higher non-specific general psychopathology--as reflected in the SCL-90-R--and significantly shorter BH duration. PMID- 17427183 TI - Mapping disability-adjusted life years: a Bayesian hierarchical model framework for burden of disease and injury assessment. AB - This paper presents a Bayesian disability-adjusted life year (DALY) methodology for spatial and spatiotemporal analyses of disease and/or injury burden. A Bayesian disease mapping model framework, which blends together spatial modelling, shared-component modelling (SCM), temporal modelling, ecological modelling, and non-linear modelling, is developed for small-area DALY estimation and inference. In particular, we develop a model framework that enables SCM as well as multivariate CAR modelling of non-fatal and fatal disease or injury rates and facilitates spline smoothing for non-linear modelling of temporal rate and risk trends. Using British Columbia (Canada) hospital admission-separation data and vital statistics mortality data on non-fatal and fatal road traffic injuries to male population age 20-39 for year 1991-2000 and for 84 local health areas and 16 health service delivery areas, spatial and spatiotemporal estimation and inference on years of life lost due to premature death, years lived with disability, and DALYs are presented. Fully Bayesian estimation and inference, with Markov chain Monte Carlo implementation, are illustrated. We present a methodological framework within which the DALY and the Bayesian disease mapping methodologies interface and intersect. Its development brings the relative importance of premature mortality and disability into the assessment of community health and health needs in order to provide reliable information and evidence for community-based public health surveillance and evaluation, disease and injury prevention, and resource provision. PMID- 17427184 TI - Identification of two risk haplotypes for schizophrenia and bipolar disorder in the synaptic vesicle monoamine transporter gene (SVMT). AB - The synaptic vesicular monoamine transporter (SVMT) plays a key role in monoaminergic neurotransmission determining the size of neurotransmitter vesicular pools available for exocytotic release. Recently, several lines of evidence have suggested that altered functions of SVMT may be involved in the pathogenesis of certain neuropsychiatric diseases, including psychotic and mood disorders. In the present study, we tested the potential involvement of SVMT gene variants in the etiology of schizophrenia and bipolar disorder. Five different SNPs (T440G, C1368T, T2666C, A2683C, and A745G) were included in the analysis covering a region of about 35 kb along the SVMT gene. Analyses were performed in a case-control sample consisting of 88 bipolar patients, 107 subjects with schizophrenia, and 164 controls. Two risk haplotypes for both schizophrenia and bipolar disorder in SVMT gene were identified. Particularly, 2666T-2683A-745G (TAG) and 2666C-2683C-745A (CCA) combinations were significantly more frequent in both bipolar and schizophrenic patients than in controls. UNPHASED package estimated haplotype effects for all patients yielded relative risks of 4.1 (95%CI: 1.83-9.21) for TAG combination and 2.336 (95%CI: 1.28-4.26) for CCA haplotype. Conversely, 2666T-2683C-745A (TCA) and 2666C-2683A-745G (CAG) haplotypes seemed to protect against these mental disorders, since the estimated frequency in control chromosomes was 12% whilst such haplotypes were not observed in any bipolar or schizophrenic subject (P < 0.0000). Our results strongly suggest that SVMT gene or certain regions of it may constitute a genetic substrate of susceptibility for both schizophrenia and bipolar disorder. PMID- 17427185 TI - Nuclear receptor NR4A2 IVS6 +18insG and brain derived neurotrophic factor (BDNF) V66M polymorphisms and risk of Taiwanese Parkinson's disease. AB - Both of environmental and genetic factors confer vulnerability to Parkinson's disease (PD). NR4A2 (Nurr1), a member of the steroid/thyroid hormone nuclear receptor superfamily, is essential for the neurogenesis and differentiation of dopaminergic neurons in the midbrain. Brain derived neurotrophic factor (BDNF) deficiency may play a role in the pathogenesis of PD, as the surviving dopaminergic nigrostriatal neurons have reduced levels of BDNF. This study examines whether BDNF V66M (c.196 G --> A) or NR4A2 IVS6 +18insG polymorphism is associated with the risk of Taiwanese PD and the age of onset using a case control study. The genotype or allele frequency distribution of both BDNF V66M and NR4A2 IVS6 +18insG polymorphisms was not significantly different between the cases and the controls. Neither BDNF nor NR4A2 polymorphism influences PD onset age. Notably, after stratification by sex, female individuals carrying the NR4A2 2G/2G genotype demonstrated a trend toward significant decrease in risk of developing PD (OR = 0.49, 95% CI = 0.25-0.96, P = 0.039). These results suggest that the NR4A2 IVS6 +18insG polymorphism may play a minor role in PD susceptibility among Taiwanese women. PMID- 17427186 TI - Association analysis of COMT polymorphisms and schizophrenia in a Chinese Han population: a case-control study. AB - Schizophrenia is a common disease with complex mode of inheritance; great efforts have been made to identify the susceptible genes. Catechol-O-methyltransferase (COMT) gene has long been considered as a candidate gene mainly because of two reasons: First, it encodes a key dopamine catabolic enzyme. Second, it maps to the velocardiofacial syndrome (VCFS) region of chromosome 22q11, which is associated with schizophrenia predisposition. Numerous case-control and family based studies have been conducted, majority of them focused on a functional Val/Met polymorphism (rs4680). Unfortunately, these studies have produced conflicting results. In a previous report, Shifman et al. found a three-marker haplotype (rs737865-rs4680-rs165599) that showed significant association with schizophrenia. In this study, we try to replicate their findings in Chinese Han population and failed to find any associations. PMID- 17427187 TI - Why do young women smoke? IV. Role of genetic variation in the dopamine transporter and lifetime traumatic experience. AB - Cigarette smoking is a complex behavior to which environmental, psychological, and genetic factors contribute. Applying a multifactorial model, we examined the role of genetic variation in the dopamine transporter (DAT1) in smoking initiation (SI) and nicotine dependence. The participants were female college students who had never smoked (n = 148) or had smoked daily for at least a year (n = 242). All participants provided extensive background information and completed a series of psychological instruments. Five SNPs were genotyped in the 3' and 5' regions of DAT1. Data were analyzed by logistic regression. The best fitting model for SI (P = 1.9 x 10(-17), Nagelkerke R2 = 0.33) revealed novelty seeking (OR = 1.14, P = 0.000004) and lifetime traumatic experience (OR = 2.3, P = 0.001) as risk factors and a DAT1_E15 + 274-DAT1_VNTR G-9 haplotype as protective (OR = 0.57, P = 0.03). In the model for nicotine dependence (P = 1.4 x 10(-8), Nagelkerke R2 = 0.27) novelty seeking was a risk factor (OR = 1.07, P = 0.03); the DAT1_E15+274-DAT1_VNTR G-9 haplotype (OR = 0.37, P = 0.001) and the interaction between trauma and a DAT1_E15 + 274-DAT1_VNTR C-9 haplotype (OR = 0.15, P = 0.01) were protective. Lifetime experience of trauma was associated with high nicotine dependence among non-carriers of the C-9 haplotype but not among carriers of this haplotype. These findings indicate that in the context of a multifactorial model, haplotypes in the 3' region of DAT1 influence the propensity of young women to initiate smoking as well as the severity of nicotine dependence once the habit is established. A haplotype in the 3' untranslated region of DAT1 modifies the effect of lifetime traumatic experience on the severity of nicotine dependence. PMID- 17427188 TI - CGG repeat length correlates with age of onset of motor signs of the fragile X associated tremor/ataxia syndrome (FXTAS). AB - Fragile X-associated tremor/ataxia syndrome (FXTAS) is a late-onset neurological disorder among carriers of premutation CGG-repeat expansions within the FMR1 gene. Principal features of FXTAS include progressive action tremor and gait ataxia, with associated features of parkinsonism, peripheral neuropathy, dysautonomia, and cognitive decline. Although both clinical and neuropathologic features of FXTAS are known to be highly associated with CGG repeat length, the relationship between repeat length and age-of-onset is not known. To address this issue, the ages of onset of action tremor and gait ataxia were documented by history for 93 male carriers. For this cohort, the mean ages of onset were 62.6 +/- 8.1 years (range, 39-78 years) for tremor, and 63.6 +/- 7.3 years (range, 47 78 years) for ataxia; the mean CGG repeat number was 88.5 +/- 14 (range, 60-133). Analysis of the relationship between clinical onset and molecular measures revealed significant correlations between CGG repeat number and onset of both tremor (P = 0.001) and ataxia (P = 0.002), as well as overall onset (P < 0.0001). Our findings indicate that the CGG repeat number is a potential predictor of the age of onset of core motor features of FXTAS. PMID- 17427189 TI - Association of the neuropilin-2 (NRP2) gene polymorphisms with autism in Chinese Han population. AB - Autism is a pervasive neurodevelopmental disorder, with a significant role of genetic factors in its development. The neuropilin-2 (NRP2) gene is localized to 2q34, an autism susceptibility locus. NRP2 has been demonstrated to both guide axons and to control neuronal migration in the central nervous system. It has been reported that NRP2 may be required in vivo for sorting migrating cortical and striatal interneurons to their correct destination. We examine the association between the NRP2 gene and autism using a cohort of 169 Chinese Han family trios. Four single nucleotide polymorphisms (SNPs) were genotyped by the polymerase chain reaction-based restriction fragment length polymorphism (PCR RFLP) analyses. The transmission disequilibrium tests (TDT) of SNPs and haplotype association were carried out using the TDTPHASE program. We found significant genetic association between autism and two of the SNPs of the NRP2 gene (rs849578: P = 0.017, rs849563: P = 0.027), as well as specific haplotypes, especially those formed by rs849563. Furthermore, haplotypes constructed with all markers showed significant excess transmission in both global and individual haplotype analyses (P = 0.004 and 0.017, respectively). The polymorphisms in the NRP2 gene are associated with autism, implying that the NRP2 gene may render individuals to be predisposed to autism. PMID- 17427190 TI - Association studies testing for risk for late-onset Alzheimer's disease with common variants in the beta-amyloid precursor protein (APP). AB - Linkage studies have suggested a susceptibility locus for late-onset Alzheimer's disease (LOAD) on chromosome 21. A functional candidate gene in this region is the beta-amyloid precursor protein (APP) gene. Previously, coding mutations in APP have been associated with early onset Alzheimer's Disease (EOAD). Three copies of APP are associated with AD pathology in Down's syndrome and in EOAD, suggesting that overexpression of APP may be a risk factor for LOAD. Although APP is a strong functional and positional candidate, to date there has been no thorough investigation using a dense map of SNPs across the APP gene. In order to investigate the role of common variation in the APP gene in the risk of LOAD, we genotyped 44 SNPs, spanning 300 kb spanning the entire gene, in a large case control series of 738 AD cases and 657 healthy controls. The SNPs showed no association in genotypic or allelic tests, even after stratification for presence or absence of the APOE 4 allele. Haplotype analysis also failed to reveal significant association with any common haplotypes. These results suggest that common variation in the APP gene is not a significant risk factor for LOAD. However, we cannot rule out the possibility that multiple rare variants that increase APP expression or Abeta production might influence the risk for LOAD. PMID- 17427191 TI - Low brain-derived neurotrophic factor (BDNF) levels in serum of Huntington's disease patients. AB - Huntington's disease (HD) is a neurodegenerative disorder characterized by motor, cognitive, and psychiatric symptoms and by a progressive degeneration of neurons in basal ganglia and in brain cortex. Brain-derived neurotrophic factor (BDNF) is a pro-survival factor for striatal neurons. Some evidence implicates a brain BDNF deficiency, related to mutated huntingtin expression, in the selective vulnerability of striatal neurons in HD. We compared BDNF serum levels in 42 patients with HD (range 28-72 years, mean age 51.9 +/- 11.5), and 42 age-matched healthy subjects (range 25-68 years, mean age 48.2 +/- 12.5). We evaluated the potential relationship between BDNF serum levels, CAG repeat number (range 40-54, mean 44.8 +/- 3.4) and duration of illness (range 6-228 months, mean 103.6 +/- 62.1). Serum BDNF levels were significantly lower in patients than in age-matched healthy subjects. Lower BDNF levels were associated with a longer CAG repeat length and a longer duration of illness. Severity of the illness, as assessed by the Unified Huntington's Disease Rating Scale (UHDRS) motor and cognitive scores, was negatively related to serum BDNF levels. These results in vivo confirm that the huntingtin mutation causes BDNF production to decline and show that the BDNF deficiency is detectable in HD patients' sera. Further studies on a larger sample size should confirm whether BDNF concentrations in patients' serum could be a useful clinical marker related to the patients' disease phenotype. PMID- 17427192 TI - Sustained attention and response inhibition in boys with fragile X syndrome: measures of continuous performance. AB - Sustained attention and response inhibition were examined in boys with full mutation fragile X syndrome (FXS) using adapted visual and auditory continuous performance tests (CPTs). Only 61% of 56 boys with visual CPT data and 54% of 52 boys with auditory data were able to demonstrate sufficient understanding to complete the visual and auditory CPTs, respectively. Mental age (MA) predicted whether boys with FXS were able to demonstrate understanding of the CPTs. The performance of boys with FXS who were able to complete the CPTs was compared to a sample of boys without disabilities matched on MA. Boys with FXS demonstrated similar or smaller declines in sustained attention over task time than their MA matched peers on the visual and auditory CPTs, respectively, but consistently demonstrated greater declines in response inhibition over task time than their MA matched peers. There were no differences between groups for response time of hits. Higher MAs consistently predicted better sustained attention and response inhibition over task time on the visual and auditory CPTs. Furthermore, boys taking psychotropic medication performed better at the beginning of most tasks, although their performance deteriorated at a faster rate over time, and boys rated as meeting diagnostic criteria for ADHD-hyperactive type had more difficulty over task time with response inhibition on the auditory CPT. For both boys with FXS and their MA matches, performance was better on the visual CPT than on the auditory CPT though this effect may be attributable to a number of factors other than the modality. PMID- 17427193 TI - MECP2 coding sequence and 3'UTR variation in 172 unrelated autistic patients. AB - Mutations in the coding sequence of the methyl-CpG-binding protein 2 gene (MECP2), which cause Rett syndrome (RTT), have been found in male and female autistic subjects without, however, a causal relation having unequivocally been established. In this study, the MECP2 gene was scanned in a Portuguese autistic population, hypothesizing that the phenotypic spectrum of mutations extends beyond the traditional diagnosis of RTT and X-linked mental retardation, leading to a non-lethal phenotype in male autistic patients. The coding region, exon intron boundaries, and the whole 3'UTR were scanned in 172 patients and 143 controls, by Detection of Virtually All Mutations-SSCP (DOVAM-S). Exon 1 was sequenced in 103 patients. We report 15 novel variants, not found in controls: one missense, two intronic, and 12 in the 3'UTR (seven in conserved nucleotides). The novel missense change, c.617G > C (p.G206A), was present in one autistic male with severe mental retardation and absence of language, and segregates in his maternal family. This change is located in a highly conserved residue within a region involved in an alternative transcriptional repression pathway, and likely alters the secondary structure of the MeCP2 protein. It is therefore plausible that it leads to a functional modification of MeCP2. MECP2 mRNA levels measured in four patients with 3'UTR conserved changes were below the control range, suggesting an alteration in the stability of the transcripts. Our results suggest that MECP2 can play a role in autism etiology, although very rarely, supporting the notion that MECP2 mutations underlie several neurodevelopmental disorders. PMID- 17427194 TI - Investigation of parent-of-origin effects in ADHD candidate genes. AB - Attention deficit hyperactivity disorder (ADHD) is a common early-onset childhood disorder with a strong genetic component. Results from previous studies have suggested that there may be a parent-of-origin effect for ADHD candidate genes. In particular, a recent investigation identified a pattern of paternal over transmission of risk alleles for nine ADHD candidate genes. We examined this phenomenon in a sample of 291 trios for five genes previously associated with ADHD (HTR1B, SNAP-25, DRD5, DAT1, and BDNF). Using a dense map of markers and two analytic methods in this relatively large family-based sample, we do not find any evidence for significant paternal over-transmission of risk alleles in these candidate loci. Thus, we conclude that a substantial parent-of-origin effect is unlikely for these leading ADHD candidate genes. PMID- 17427196 TI - Monoamine oxidase A gene polymorphism predicts adolescent outcome of attention deficit/hyperactivity disorder. AB - ADHD is generally deemed to be a highly heritable disorder with mean heritability of 0.75. The enzyme monoamine oxidase (MAO), which has both A and B types, has long been considered a candidate pathological substrate for ADHD, and more recently, the genes for both MAO enzymes have been examined as mediators of the illness. Previous studies indicated that 30-50% of children with ADHD will experience symptoms that persist into adolescence and will have more significant impairment in social and neuropsychological functioning compared to those whose symptoms have remitted. Genes may also influence these characteristics of the disorder, and in this context MAO genes may also be candidates for moderating the presentation of ADHD. The current study examined the association between adolescent outcome of ADHD and MAO gene polymorphisms, including the 941T > G polymorphism in exon 8 (rs1799835) and 1460C > T polymorphism in exon 14 (rs1137070) of the MAOA gene, and the A > G polymorphism in intron13 (rs1799836), C > T polymorphism in the 3'UTR (rs1040399), and 2327T > C polymorphism in exon15 of the MAOB gene. Significant associations were observed between the MAOA gene polymorphisms and ADHD remission. Due to the small sample size and the possibility of phenotypic and etiologic heterogeneity of ADHD outcomes across ethnic or geographic groups, these results must be replicated before they can be generalized to other populations. PMID- 17427195 TI - Mutation screening of the PTEN gene in patients with autism spectrum disorders and macrocephaly. AB - Mutations in the PTEN gene are associated with a broad spectrum of disorders, including Cowden syndrome (CS), Bannayan-Riley-Ruvalcaba syndrome, Proteus syndrome, and Lhermitte-Duclos disease. In addition, PTEN mutations have been described in a few patients with autism spectrum disorders (ASDs) and macrocephaly. In this study, we screened the PTEN gene for mutations and deletions in 88 patients with ASDs and macrocephaly (defined as >or=2 SD above the mean). Mutation analysis was performed by direct sequencing of all exons and flanking regions, as well as the promoter region. Dosage analysis of PTEN was carried out using multiplex ligation-dependent probe amplification (MLPA). No partial or whole gene deletions were observed. We identified a de novo missense mutation (D326N) in a highly conserved amino acid in a 5-year-old boy with autism, mental retardation, language delay, extreme macrocephaly (+9.6 SD) and polydactyly of both feet. Polydactyly has previously been described in two patients with Lhermitte-Duclos disease and CS and is thus likely to be a rare sign of PTEN mutations. Our findings suggest that PTEN mutations are a relatively infrequent cause of ASDs with macrocephaly. Screening of PTEN mutations is warranted in patients with autism and pronounced macrocephaly, even in the absence of other features of PTEN-related tumor syndromes. PMID- 17427197 TI - D-glucose stimulation of L-arginine transport and nitric oxide synthesis results from activation of mitogen-activated protein kinases p42/44 and Smad2 requiring functional type II TGF-beta receptors in human umbilical vein endothelium. AB - Elevated extracellular D-glucose increases transforming growth factor beta1 (TGF beta1) release from human umbilical vein endothelium (HUVEC). TGF-beta1, via TGF beta receptors I (TbetaRI) and TbetaRII, activates Smad2 and mitogen-activated protein kinases p44 and p42 (p42/44(mapk)). We studied whether D-glucose stimulation of L-arginine transport and nitric oxide synthesis involves TGF-beta1 in primary cultures of HUVEC. TGF-beta1 release was higher ( approximately 1.6 fold) in 25 mM (high) compared with 5 mM (normal) D-glucose. TGF-beta1 increases L-arginine transport (half maximal effect approximately 1.6 ng/ml) in normal D glucose, but did not alter high D-glucose-increased L-arginine transport. TGF beta1 and high D-glucose increased hCAT-1 mRNA expression ( approximately 8-fold) and maximal transport velocity (V(max)), L-[(3)H]citrulline formation from L [(3)H]arginine (index of NO synthesis) and endothelial NO synthase (eNOS) protein abundance, but did not alter eNOS phosphorylation. TGF-beta1 and high D-glucose increased p42/44(mapk) and Smad2 phosphorylation, an effect blocked by PD-98059 (MEK1/2 inhibitor). However, TGF-beta1 and high D-glucose were ineffective in cells expressing a truncated, negative dominant TbetaRII. High D-glucose increases L-arginine transport and eNOS expression following TbetaRII activation by TGF-beta1 involving p42/44(mapk) and Smad2 in HUVEC. Thus, TGF-beta1 could play a crucial role under conditions of hyperglycemia, such as gestational diabetes mellitus, which is associated with fetal endothelial dysfunction. PMID- 17427198 TI - AND-34/BCAR3 differs from other NSP homologs in induction of anti-estrogen resistance, cyclin D1 promoter activation and altered breast cancer cell morphology. AB - Over-expression of AND-34/BCAR3/NSP2 (BCAR3) or its binding-partner p130Cas/BCAR1 generates anti-estrogen resistance in human breast cancer lines. Here, we have compared BCAR3 to two related homologs, NSP1 and NSP3/CHAT/SHEP, with regards to expression, anti-estrogen resistance, and signaling. BCAR3 is expressed at higher levels in ERalpha-negative, mesenchymal, than in ERalpha-positive, epithelial, breast cancer cell lines. Characterization of "intermediate" epithelial-like cell lines with variable ER-alpha expression reveals that BCAR3 expression correlates with both mesenchymal and ERalpha-negative phenotypes. Levels of the BCAR3/p130Cas complex correlate more strongly with the ERalpha-negative, mesenchymal phenotype than levels of either protein alone. NSP1 and NSP3 are expressed at lower levels than BCAR3 and without correlation to ERalpha/mesenchymal status. Among NSP-transfectants, only BCAR3 transfectants induce anti-estrogen resistance and augment transcription of cyclin D1 promoter constructs. Over-expression of all homologs results in activation of Rac, Cdc42 and Akt, suggesting that these signals are insufficient to induce anti-estrogen resistance. BCAR3 but not NSP1 nor NSP3 transfectants show altered morphology, transitioning from polygonal cell groups to rounded, single cells with numerous blebs. Whereas stable over-expression of BCAR3 in MCF-7 cells does not lead to classic epithelial-to-mesenchymal transition, it does result in down-regulation of cadherin-mediated adhesion and augmentation of fibronectin expression. These studies suggest that BCAR's ability to induce anti-estrogen resistance is greater than that of other NSP homologs and may result from altered interaction of breast cancer cells with each other and the extracellular matrix. PMID- 17427199 TI - MEK/ERK-dependent uPAR expression is required for motility via phosphorylation of P70S6K in human hepatocarcinoma cells. AB - Motility and invasiveness events require specific intracellular signaling cascade activations. In cancer liver cells, one of these mechanisms could involve the MAPK MEK/ERK cascade activation which has been shown over expressed and activated in hepatocellular carcinoma. To study whether the MEK/ERK cascade is involved in the motility of HCC, we examined the effect of MEK inhibitor and ERK2 silencing using monolayer wound-healing assays and fluoroblock invasion systems. Evidence was provided that the MAPK cascade is a key transduction pathway which controls HCC cells motility and invasiveness. We could disconnect proliferation to motility using mitomycin C and we established that RNAi-mediated inhibition of ERK2 led to strongly reduced cell motility. To improve our understanding, we analysed the regulation and the role of urokinase receptor (uPAR) in this process. We provided evidence that uPAR was under a MEK/ERK dependent mechanism and blocking uPAR activity using specific antagonist or inhibiting its expression by RNA interference which resulted in complete inhibition of motility. Moreover, we found in MAPK inhibited cultures and in uPAR silencing cells that p70S6K phosphorylation on residue Thr-389 was significantly reduced, whereas Ser-421/Thr 424 phosphorylation did not change. We highlighted that the FRAP/mTOR pathway did not affect motility and Thr-389 phosphorylation. Furthermore, we demonstrated that p70S6K inhibition by RNA interference completely inhibited hepatocarcinoma cell motility. Therefore, targeting uPAR and/or MEK/ERK/S6K by RNA interference could be a major therapeutic strategy for the future treatment of invasive hepatocarcinoma cells. PMID- 17427200 TI - Nanoparticulate delivery of suicide DNA to murine prostate and prostate tumors. AB - BACKGROUND: Currently available treatments for benign prostatic hyperplasia (BPH) and localized prostate cancer are generally effective but are often attended by serious side effects that impact on the quality of life. In particular, most current therapies are non-specific, with surgery, radiation, and chemical ablation having the potential to cause damage to surrounding tissue. Here, we demonstrate the effectiveness of a prostate-specific, locally delivered gene therapy for the targeted killing of prostate cells. METHODS: Using a degradable, poly(beta-amino ester) polymer, poly(butane diol diacrylate co amino pentanol) (C32), we developed a nanoparticulate system to deliver a diphtheria toxin suicide gene (DT-A) driven by a prostate specific promoter to cells. These C32/DT A nanoparticles were directly injected to the normal prostate and to prostate tumors in mice. RESULTS: Nearly 50% of normal prostates showed a significant reduction in size, attributable to cellular apoptosis, whereas injection with naked DT-A-encoding DNA had little effect. Significant apoptosis was also observed in C32/DT-A injected prostate tumors. Importantly, no damage to surrounding tissue was observed. CONCLUSIONS: These results suggest that local delivery of poly(beta-amino ester) polymer/ DT-A nanoparticles may have application in the treatment of BPH and prostate cancer. PMID- 17427201 TI - Work postures and neck-shoulder pain among orchestra musicians. AB - BACKGROUND: The purpose of the present study was to identify associations between a work posture with elevated arm position, duration of active playing time, and neck-shoulder pain among orchestra musicians. METHODS: In this study, with a cross-sectional study design, a total of 235 subjects from 12 Swedish orchestras were categorized into four exposure groups according to arm position and duration of active playing time. Logistic regression analysis was used to calculate odds ratios (OR) for neck-shoulder pain in the four exposure groups. RESULTS: A higher prevalence of neck-shoulder pain were found in the groups "elevated arm position, <2 hr per workday" [OR 4.15 (1.30-13.22)], and "elevated arm position, >3 hr per workday" [OR 5.35 (1.96-14.62)] compared to the group "neutral arm position, <2 hr per workday". CONCLUSIONS: Musicians working in an elevated arm position (e.g., violinists, violists, flutists, and trumpet players) had a higher prevalence of neck-shoulder pain than those working in a more neutral position. PMID- 17427202 TI - Registry-based case-control study of cancer in California firefighters. AB - BACKGROUND: There is no consensus whether firefighters are at increased cancer risk for particular cancers. Previous studies have been small, mostly investigated cancer mortality, and suggested increased risks for brain, bladder, testicular, prostate, thyroid and colo-rectal cancers, leukemia, and melanoma. METHODS: Records of all male cancers registered in California during 1988-2003 were obtained. Firefighters were identified from occupation and industry text fields. Logistic regression analysis used other cancers as controls. RESULTS: Of the 804,000 eligible records, 3,659 had firefighting as their occupation. Firefighting was associated with testicular cancer (odds ratio = 1.54, 95% confidence interval: 1.18-2.02), melanoma (1.50, 1.33-1.70), brain cancer (1.35, 1.06-1.72), esophageal cancer (1.48, 1.14-1.91), and prostate cancer (1.22, 1.12 1.33). CONCLUSIONS: Use of other-cancer controls and lack of an occupational history may have biased relative risks towards the null. However, this study, which contained more firefighter cancers than any previous epidemiologic study, produced evidence supporting some prior hypotheses. PMID- 17427203 TI - A genome-wide linkage analysis for the personality trait neuroticism in the Irish affected sib-pair study of alcohol dependence. AB - Neuroticism is a personality trait which reflects individual differences in emotional stability and vulnerability to stress and anxiety. Consistent evidence shows substantial genetic influences on variation in this trait. The present study seeks to identify regions containing susceptibility loci for neuroticism using a selected sib-pair sample from Ireland. Using Merlin regress, we conducted a 4 cM whole-genome linkage analysis on 714 sib-pairs. Evidence for linkage to neuroticism was found on chromosomes 11p, 12q, and 15q. The highest linkage peak was on 12q at marker D12S1638 with a Lod score of 2.13 (-log p = 2.76, empirical P-value <0.001). Our data also support gender specific loci for neuroticism, with male specific linkage regions on chromosomes 1, 4, 11, 12, 15, 16, and 22, and female specific linkage regions on chromosomes 2, 4, 9, 12, 13, and 18. Some genome regions reported in the present study replicate findings from previous linkage studies of neuroticism. These results, together with prior studies, indicate several potential regions for quantitative trait loci for neuroticism that warrant further study. PMID- 17427204 TI - Bayesian and frequentist two-stage treatment strategies based on sequential failure times subject to interval censoring. AB - For many diseases, therapy involves multiple stages, with the treatment in each stage chosen adaptively based on the patient's current disease status and history of previous treatments and clinical outcomes. Physicians routinely use such multi stage treatment strategies, also called dynamic treatment regimes or treatment policies. We present a Bayesian framework for a clinical trial comparing two stage strategies based on the time to overall failure, defined as either second disease worsening or discontinuation of therapy. Each patient is randomized among a set of treatments at enrollment, and if disease worsening occurs the patient is then re-randomized among a set of treatments excluding the treatment received initially. The goal is to select the two-stage strategy having the largest average overall failure time. A parametric model is formulated to account for non constant failure time hazards, regression of the second failure time on the patient's first worsening time, and the complications that the failure time in either stage may be interval censored and there may be a delay between first worsening and the start of the second stage of therapy. Four different criteria, two Bayesian and two frequentist, for selecting a best strategy are considered. The methods are applied to a trial comparing two-stage strategies for treating metastatic renal cancer, and a simulation study in the context of this trial is presented. Advantages and disadvantages of this design compared to standard methods are discussed. PMID- 17427206 TI - Renal revascularization in diabetics: few facts, firm convictions, forward momentum. PMID- 17427207 TI - Reproducible coronary plaque quantification by multislice computed tomography. AB - BACKGROUND: The aim of this study was to investigate reproducibility and accuracy of computer-assisted coronary plaque measurements by multislice computed tomography coronary angiography (QMSCT-CA). METHODS AND RESULTS: Forty-eight patients undergoing MSCT-CA and coronary arteriography for symptomatic coronary artery disease and quantitative intravascular ultrasound (IVUS, QCU) were examined. Two investigators performed the QMSCT-CA twice and a third investigator performed the QCU, all blinded for each other's results. There was no difference found for the matched region of interest (ROI) lengths (QCU 29.4 +/- 13 mm vs. QMSCT-CA 29.6 +/- 13 mm, P = 0.6; total length = 1,400 mm). The comparison of volumetric measurements showed (lumen QCU 267 +/- 139 mm(3) vs. mean QMSCT-CA 177 +/- 91 mm(3), P < 0.001; vessel 454 +/- 194 mm(3) vs. 398 +/- 187 mm(3), P <<0.001; and plaque 189 +/- 93 mm(3) vs. 222 +/- 121 mm(3); investigator 1, P = 0.02; and investigator 2, P = 0.07) significant differences. Automated lumen detection was also applied for QMSCT-CA (218 +/- 112 mm(3), P < 0.001 vs. QCU). The interinvestigator variability measurements for QMSCT-CA showed no significant differences. CONCLUSION: QMSCT-CA systematically underestimates absolute coronary lumen- and vessel dimensions when compared with QCU. However, repeated measurements of coronary plaque by QMSCT-CA showed no statistically significant differences, although, the outcome showed a scattered result. Automated lumen detection for QMSCT-CA showed improved results when compared with those of human investigators. PMID- 17427208 TI - Passing the torch: challenges ahead. PMID- 17427209 TI - Abnormal cortical activation during response inhibition in 22q11.2 deletion syndrome. AB - 22q11.2 deletion syndrome (22q11.2DS) is a well-known genetic risk factor for schizophrenia. The catechol-O-methyltransferase (COMT) gene falls within the 22q11.2 minimal critical region of the deletion. Brain activity, as measured by functional magnetic resonance imaging (fMRI) during a Go/NoGo, response inhibition task was assessed in adolescents with 22q11.2DS (n = 13), typically developing (TD) controls (n = 14), and controls with developmental disability (DD, n = 9). Subjects with 22q11.2DS were also genotyped for the COMT Met/Val polymorphism. Groups did not differ on task performance. However, compared to both control groups, the 22q11.2DS group showed greater brain activation within left parietal regions. Comparison of brain activation between 22q11.2DS Met and Val subgroups revealed significantly increased activation (Met>Val) in the cingulate but not the dorsolateral prefrontal cortex. These preliminary findings suggest that adolescents with 22q11.2DS compensate for executive dysfunction via recruitment of parietal regions. Further, the COMT Met subgroup of 22q11.2DS may recruit additional cingulate activation for tasks requiring attention and inhibition. 22q11.2DS is a unique model for learning about the deleterious effects of decreased dosage of the COMT gene on brain function. PMID- 17427210 TI - A metastatic renal carcinoid tumor presenting as breast mass: a diagnostic dilemma. AB - We present clinicopathological and cytological findings of a well-defined breast mass in a patient with history of primary renal carcinoid tumor. Fine-needle aspiration (FNA) cytology showed monotonous tumor cells with plasmacytoid appearance arranged singly and in small clusters. Occasional tumor cells were arranged in acinar architecture resembling glandular differentiation. Tumor cells showed fine speckled chromatin. The unusual location for metastasis of this rare type of carcinoid tumor and overlapping cytological features with primary mammary carcinoma led to an erroneous preliminary cytological diagnosis of primary breast carcinoma with plasmacytoid features. Tumor cells in the corresponding cell block showed strong diffuse positivity for synapthophysin and pan-cytokeratin with weak focal positivity for chromogranin markers. These patterns of immunostaining were similar to the original renal carcinoid tumor. To the best of our knowledge, a few cases of carcinoid tumor metastatic to the breast have been reported in the literature and more than half of these cases were initially misdiagnosed as primary breast carcinoma causing unnecessary surgical treatment. This is a first reported case of metastatic renal carcinoid tumor into breast diagnosed with FNA biopsy. This report highlights the cytological features of well-differentiated neuroendocrine tumor (carcinoid tumor) and its potential diagnostic pitfalls. PMID- 17427211 TI - Acardiac syndrome coexisting with complex skeletal dysplasia in the co-twin: 3D sonographic findings. AB - We present a case of twin reversed arterial perfusion syndrome complicated by an acardius acephalus fetus and complex skeletal dysplasia in the co-twin, including a reduction of the right arm and absence of the right scapula and 4 ribs, as evaluated with three-dimensional sonography and MRI. PMID- 17427212 TI - Gonadal brucellar abscess: imaging and clinical findings in 3 cases and review of the literature. AB - We report the cases of 3 patients with brucellar gonadal abscess who were examined with gray-scale and color Doppler sonography. In these 3 cases, sonography revealed a thick-walled avarian (n = 2) or testicular (n = 1) abscess. Duplex Doppler sonography indicated a low-resistance type of flow in all 3 patients. The diagnosis of brucellar gonadal abscess is difficult, because it may have a misleading tumor-like appearance. Awareness of the imaging findings of gonadal abscess in patients with brucellosis and correlation with the results of serologic testing can allow early diagnosis, especially in endemic areas. PMID- 17427213 TI - Reduced flow velocity in the internal carotid artery independently of cardiac hemodynamics in patients with cerebral ischemia. AB - PURPOSE: To retrospectively investigate the relationships between carotid flow velocities, clinical features and cardiac hemodynamics to assess the meaning and significance of reduced carotid flow velocities in patients with cerebral ischemic symptoms. METHODS: We selected the files from patients who had undergone duplex Doppler sonographic examination of extracranial carotid arteries, echocardiography, and MR angiography, and in whom the following parameters were available: peak systolic (PSV) and end diastolic (EDV) flow velocity, pulsatility index (PI), and diameter of the left and right common (CCA) and internal (ICA) carotid arteries, intima-media thickness (IMT) of the left and right CCA, left ventricle (LV) mass, peak flow velocity on LV outflow tract, and fractional shortening (FS). Patients with stenosis of the carotid artery or its main intracranial branches were excluded, as were patients with major cerebral infarction, severe intracranial abnormality, or heart function disorder. The remaining 59 patients were subdivided according to the presence or absence of cerebral ischemic symptoms, diabetes mellitus, arterial hypertension, and hyperlipidemia for multivariate analyses and stepwise regressions. RESULTS: Women had smaller diameters and lower PI in the left and right CCA, and smaller LV mass than men. Age, CCA diameter, and IMT showed an inverse correlation with carotid flow velocities in several arterial segments. There was a positive correlation between PSV in the left CCA and ICA and FS, and between PSV in the left CCA and peak velocity on LV outflow tract. Flow velocities in the left and right ICA were significantly slower in patients with than in patients without cerebral ischemic symptoms. CONCLUSIONS: Cardiac hemodynamics and carotid flow velocities are significantly related, only on the left side, probably due to larger hemodynamic stress. Increased intracerebral circulatory resistance is probably involved in the decrease in carotid flow velocity and increase in PI in patients with cerebral ischemic symptoms. PMID- 17427214 TI - Ultrasound biomicroscopy for estimation of tumor thickness for conjunctival melanoma. AB - We assessed the feasibility of using high-frequency ultrasound biomicroscopy (UBM) in estimating thickness of conjunctival melanomas preoperatively. Three patients underwent UBM prior to excision of conjunctival melanoma between February 2003 and March 2006. Conjunctival tumor thickness was estimated via UBM in each case and compared with the estimated histopathologic tumor thickness (Breslow thickness). Patient 1 had an estimated thickness of 1.99 mm via UBM versus a Breslow thickness measurement of 1.5 mm based on the surgical specimen. Patient 2 had a UBM thickness measurement of 2.4 mm and a Breslow thickness of 2.23 mm based on the surgical specimen. In patient 3, both UBM thickness and Breslow thickness were 2.3 mm. The use of high-frequency UBM is feasible for the estimation of tumor thickness for conjunctival melanomas. UBM may serve as an additional diagnostic tool for estimating tumor thickness before surgical resection of conjunctival melanoma. PMID- 17427215 TI - Relationship between cartilage stiffness and dGEMRIC index: correlation and prediction. AB - We sought to determine if a generalized relationship between the dGEMRIC index (T1Gd relaxation time) and compressive stiffness of articular cartilage could be defined across multiple samples. Osteochondral blocks were cut from 12 human tibial plateaus, six from cadaveric sources and six from total knee replacement surgeries. Each block contained submeniscal ("covered") and extrameniscal ("uncovered") cartilage regions. At approximately 18 sites per block, a pulse indentation was applied and local dGEMRIC index was determined using dGEMRIC MRI. No correlation was found between stiffness and full cartilage depth dGEMRIC index. When averaged over the depth comparable to the indentation, good correlations were found between stiffness and the dGEMRIC index whether all data were combined, or whether each sample/region was considered independently. However, the standard error of the estimate for predicting stiffness from the local dGEMRIC index was improved when the uncovered and covered regions were considered separately. Further improvement in predictive capacity was obtained if, rather than predict absolute stiffness, differences in the dGEMRIC index across a region were used to predict difference in stiffness. The dGEMRIC index is highly correlated to mechanical stiffness. A generalized relationship was found to provide good correspondence across sources and regions. Use of the dGEMRIC index as a predictive measure of stiffness is possible, depending on the application's acceptable error. PMID- 17427216 TI - Cyto-histological correlations in primary, recurrent, and metastatic bone and soft tissue osteosarcoma. Institut Curie's experience. AB - To determine diagnostic cytomorphologic features of osteosarcoma on fine-needle aspiration materials, we reviewed the cytologic material and corresponding histologic slides of 126 tumors in 107 patients. Fifty-five (43.6%) tumors were primary, 31 (24.6%) were recurrent, and 40 (31.8%) were metastatic. Review of original cytology reports revealed that 120 (95.3%) tumors were diagnosed as malignant. Six (4.7%) cases were reported as suspicious, false-negative, or unsatisfactory samples. Our findings showed that osteoblastic roundish cells, spindle-shaped cells, reactive giant cells, and osteoid were the most consistent features representative of osteosarcoma. Periosteal reactions, fractures with callous formation, giant cells of osteoclastic type in various conditions, chondrosarcoma with enchondral ossification are entities to consider in the differential diagnosis. PMID- 17427217 TI - Endoscopic ultrasound-guided biopsies of pancreatic masses: comparison between fine needle aspirations and needle core biopsies. AB - Endoscopic ultrasound-guided fine needle aspiration (EUS-guided FNA) is a highly sensitive and specific method for diagnosing pancreatic masses. Alternatively, EUS-guided needle core biopsies (NCB) have also been introduced. We sought to determine efficacies of pancreatic EUS-guided FNAs and NCBs. Records of consecutive EUS-guided FNAs received over a 24-mo-period were reviewed. Cases with concurrent NCBs were selected for the study. The diagnoses from the two modalities were compared and designated concordant (CC) or discordant (DC). Of 252 cases, 52 had concurrent NCBs. The final diagnoses included primary and secondary tumors. Of the 52 cases, 29/52 (55.8%) were CC and 23/52 (44.2%) were DC. The sensitivities for FNAs and NCBs were 95.0% and 67.6%, respectively. Both modalities were 100% specific. Direct comparison between EUS-guided FNAs and NCBs demonstrated that the former are more sensitive for diagnosing pancreatic neoplasms, both primaries and metastases. There was no correlation between CC/DC cases and type of neoplasm. PMID- 17427218 TI - Role of FNA cytology and immunochemistry in the diagnosis and management of medullary thyroid carcinoma: report of six cases and review of the literature. AB - Medullary thyroid carcinoma (MTC) is a rare neuroendocrine thyroid malignancy. This study retrospectively reviewed 10 fine-needle aspiration samples from six MTC patients. Aspirated specimens were from thyroid (3), cervical lymph nodes (5), left lung (1), and anterior chest wall (1). Cytomorphology consisted predominantly of plasmacytoid cells (3 cases), spindle cells (2 cases), and epithelioid cells (1 case). However, all specimens had a mixture of other cell types and "salt and pepper" chromatin. Only one specimen showed Congo-red positive amyloid. Calcitonin was expressed in 7/7 specimens. Four patients underwent surgical excision and MTC was confirmed in all four. Follow-up studies included serum calcitonin (3/6 cases) and imaging (2/6 cases). One patient had MTC associated with multiple endocrine neoplasia IIA syndrome and one had familial MTC with a history of MTC in mother. In conclusion, the cytomorphology of MTC is typical and calcitonin immunostain is a reliable method for confirming primary or metastatic MTC. Early cytological diagnosis of MTC positively impacted patient management. Follow-up with serum calcitonin and imaging is helpful in the early detection of recurrences. PMID- 17427219 TI - Merkel cell carcinoma and chronic lymphocytic leukemia (collision tumor) of the arm: a diagnosis by fine-needle aspiration biopsy. AB - Simultaneous involvement of the same anatomical site by two different primary malignant tumors is rare. Cases of hematopoietic malignancies associated with breast and skin neoplasms have been described. The association of chronic lymphocytic leukemia (CLL) and Merkel cell carcinoma (MCC) has been established, although the cause for this association is still unclear. There are reports of MCC metastatic to lymph nodes involved by CLL. We report the case of a 57-year old man with history of CLL with concurrent involvement of the arm by CLL and MCC diagnosed on fine-needle aspiration biopsy (FNA). To our knowledge, this is the first reported case of such tumors colliding in a nonlymphoid site, diagnosed by FNA in the English literature. PMID- 17427220 TI - Elastofibroma dorsi. PMID- 17427221 TI - Cytomorphological correlates of epidermal growth factor receptor mutations in lung carcinoma. AB - The initial diagnosis of lung carcinoma is frequently made by fine-needle aspiration biopsy. Novel therapeutic strategies of this disease include tyrosine kinase inhibitors (TKI), such as gefitinib (Iressa) or erlotinib (Tarceva), which target the kinase domain of epidermal growth factor receptor (EGFR). Somatic mutations of this region have been shown to predict a therapeutic response of lung carcinomas to TKI. EGFR mutations have been described in adenocarcinomas of the lung, especially the bronchioloalveolar subtype, which has both cytopathologic and histopathologic definitions. This study investigates whether tumors with EGFR mutations display a characteristic phenotype on fine-needle aspiration biopsy. We identified 37 fine-needle aspiration biopsy of lung masses on which molecular analysis for EGFR mutations was available. Molecular analysis was performed on DNA isolated from formalin-fixed, paraffin-embedded, or frozen tissue from the corresponding core biopsies/cell blocks or resection specimens followed by PCR with primers for the tyrosine kinase region exons 18-24 and nucleotide sequence analysis by gel electrophoresis. Two observers who were blinded to the mutational data assessed several cytomorphological parameters. A semiquantitative analysis included predominant tissue pattern (flat or overlapping), nuclear features (nucleoli, intranuclear inclusions, grooves), cytoplasmic qualities, and extracellular material. All cases were adenocarcinomas primary in the lung. Thirteen cases showed EGFR mutations in exons 18, 19, 20, or 21 of the tyrosine kinase domain. The 24 cases negative for the relevant mutation served as the control group. Tumors with EGFR mutations were statistically more likely to demonstrate a predominantly flat, monolayer architecture (P=0.04) with nuclear inclusions (P=0.014) and the absence of macronucleoli (P=0.001). The predominance of flat monolayers in conjunction with the absence of extracellular mucin and macronucleoli indicated the presence of EGFR mutations with a positive predictive value of 69% and a negative predictive value of 92%. All four cases with extracellular mucin were negative for the examined mutations. Some of the traditional cytomorphological features of bronchioloalveolar carcinoma, i.e., flat monolayers, intranuclear inclusions, and the absence of macronucleoli, statistically correlated with the presence of mutations within the tyrosine kinase region of EGFR. Cytomorphological features could serve as an adjunctive predictive marker of response to TKIs and possibly to other new therapies in development. PMID- 17427222 TI - Myxoid liposarcoma. PMID- 17427223 TI - Fine-needle aspiration cytology of apocrine adenosis of the breast: report on three cases. AB - Apocrine adenosis is a distinctive breast lesion, which can sometimes be misdiagnosed as malignant histologically. Although its histological characteristics have been well established, there are limited descriptions of its cytological features. We report fine-needle aspiration (FNA) cytology of three cases of apocrine adenosis. There were diverse cytological features between cases, and in two cases, FNA smears were highly cellular. Naked nuclei were present in all cases, and were numerous in one. Enlarged nucleoli and anisonucleosis were common. Apocrine metaplasia with nuclear atypia mimicking apocrine carcinoma was prominent in one case, whereas one case lacked definite apocrine features. Regardless of their various atypical FNA cytologies, the nuclei were consistently less hyperchromatic in all cases. Although FNA cytology of apocrine adenosis has the potential to be misinterpreted as malignant, the naked nuclei of background and less hyperchromatic nuclear features may be useful in distinguishing apocrine adenosis from carcinoma. Being familiar with these distinctive breast lesions is also important in decreasing false positives. PMID- 17427224 TI - Cytomorphology and B72.3 labeling of benign and malignant ductal epithelium in pancreatic lesions compared to gastrointestinal epithelium. AB - Endoscopic ultrasound-guided pancreatic fine-needle aspiration biopsy very frequently produces gastrointestinal epithelial contamination (GIC). We studied the cytomorphology and B72.3 immunoreactivity of lesional epithelium of benign and malignant ductal lesions of the pancreas and compared the findings to our previously established template of GIC. Air-dried smears, fixed smears, and ThinPrep (TP) specimens were obtained using a cytobrush, directly from benign and malignant ductal lesions of 18 Whipple specimens, to ensure purity of the epithelium studied. Smear background, cell architecture, and cellular features were analyzed. Immunocytochemical staining with B72.3 was performed in 14 cases. Epithelium of ductal carcinoma was distinguished from benign ductal epithelium in chronic pancreatitis and GIC primarily by crowded architecture and atypical cellular features, including high nuclear-to-cytoplasmic ratio, irregular nuclei, nucleoli, and vacuolated cytoplasm. Benign ductal and GIC epithelium were only distinguished by architecture (goblet cells and brush borders), but not consistently, especially gastric epithelium that lacked these features. B72.3 shows promise in the differentiation between GIC and benign and malignant ductal epithelium, with no staining supporting benign ductal cells, fine punctate perinuclear staining correlating with GIC, and strong cytoplasmic staining supporting malignancy. PMID- 17427225 TI - Cell blocks of breast FNAs frequently allow diagnosis of invasion or histological classification of proliferative changes. AB - Two major limitations of breast fine needle aspiration (FNA) compared with core needle biopsies (CNB) are the inability to determine whether a cancer is invasive and to classify proliferative lesions. We studied 40 consecutive "rapid cell blocks" from breast FNAs with surgical pathology follow-up to test whether cell blocks can overcome these limitations. Of 25 carcinomas, invasion could be identified in the cell block sections in 11 (44%). One cystosarcoma phyllodes was suspected based on the cell block sections. Cell blocks from 12 of 14 benign breast FNAs showed sufficient cells to assign a histologic diagnosis of no hyperplasia (1 case, confirmed on follow-up) and usual hyperplasia (11 cases; confirmed in eight of 11 on follow-up). Specific histologic diagnoses included intraductal papilloma (2 cases), and in situ lobular neoplasia (2 cases). Cell blocks complement smears and monolayers and appear to overcome major limitations of breast FNA. PMID- 17427226 TI - Percutaneous coronary artery fistula closure in adults: technical and procedural aspects. AB - BACKGROUND: Coronary artery fistulae (CAF) are an uncommon congenital anomaly characterized by an abnormal connection between the coronary arteries and cardiac chambers or vessels without traversing the usual capillary network. CAF are associated with a number of well-described symptomatic sequelae, which may necessitate treatment. With a tendency for symptoms to develop over time, symptomatic CAF may present for the first time in adulthood, with limited data existing on the technical aspects related to transcatheter fistula closure in adults. METHODS: We describe our experience in percutaneous closure of CAF in an exclusively adult population, with an emphasis on the various procedural features of device closure in this patient group. RESULTS: Attempted transcatheter CAF closure was performed in 14 patients between 1990 and 2006. Procedural success was achieved in 11 patients, with vessel tortuosity and lumen caliber important limitations in occlusion device delivery. Procedural complications included vessel dissection and device embolization. CONCLUSION: Transcatheter occlusion is a safe and effective method of therapy for symptomatic adults with CAF. PMID- 17427227 TI - Detection of gene duplication signals of Ig folds from their amino acid sequences. AB - Protein folds may evolve from short peptide ancestors via gene duplication and fusion. For proteins with internal structural symmetry, this means that their sequences should be made up of identical repeats. However, many of these repeat signals can only be seen at the structural level yet. Motivated by the fact that proteins may have similar structures if their sequences have more than 25% identical amino acids, we suggest a method to detect the sequence repeats of proteins directly from their sequences. Using this method, we show that the internal repetitions of the immunoglobulin folds could be identified directly at the sequence level. PMID- 17427229 TI - Stepwise prediction of conformational discontinuous B-cell epitopes using the Mapitope algorithm. AB - Mapping the epitope of an antibody is of great interest, since it contributes much to our understanding of the mechanisms of molecular recognition and provides the basis for rational vaccine design. Here we present Mapitope, a computer algorithm for epitope mapping. The algorithm input is a set of affinity isolated peptides obtained by screening phage display peptide-libraries with the antibody of interest. The output is usually 1-3 epitope candidates on the surface of the atomic structure of the antigen. We have systematically tested the performance of Mapitope by assessing the effect of the algorithm parameters on the final prediction. Thus, we have examined the effect of the statistical threshold (ST) parameter, relating to the frequency distribution and enrichment of amino acid pairs from the isolated peptides and the D (distance) and E (exposure) parameters which relate to the physical parameters of the antigen. Two model systems were analyzed in which the antibody of interest had previously been co-crystallized with the antigen and thus the epitope is a given. The Mapitope algorithm successfully predicted the epitopes in both models. Accordingly, we formulated a stepwise paradigm for the prediction of discontinuous conformational epitopes using peptides obtained from screening phage display libraries. We applied this paradigm to successfully predict the epitope of the Trastuzumab antibody on the surface of the Her-2/neu receptor in a third model system. PMID- 17427230 TI - Beta-thalassemia major births after National Screening Program in Taiwan. AB - OBJECTIVE: A National Thalassemia Screening Program was adopted in Taiwan in 1993. This report examined that program's results and impact. METHODS: Patients with beta-thalassemia major born between 1994 and 2003 were recruited through the help of all thalassemia clinics in Taiwan. A structured questionnaire was designed to collect the reasons for affected births. RESULTS: There were 97 affected births from 1994 to 2003.These births resulted after informed choice (n = 4), screening problems (n = 83), and undetermined causes (n = 10). Approximately 83% (5/6) of affected births in 2003 came from interracial marriages. CONCLUSIONS: This report has identified several areas that might improve the thalassemia-screening program, including carrier screening in high school rather than in early pregnancy and the involvement of genetic counselors, providing care of new female immigrants. PMID- 17427231 TI - Computational proteomics analysis of HIV-1 protease interactome. AB - HIV-1 protease is a small homodimeric enzyme that ensures maturation of HIV virions by cleaving the viral precursor Gag and Gag-Pol polyproteins into structural and functional elements. The cleavage sites in the viral polyproteins share neither sequence homology nor binding motif and the specificity of the HIV 1 protease is therefore only partially understood. Using an extensive data set collected from 16 years of HIV proteome research we have here created a general and predictive rule-based model for HIV-1 protease specificity based on rough sets. We demonstrate that HIV-1 protease specificity is much more complex than previously anticipated, which cannot be defined based solely on the amino acids at the substrate's scissile bond or by any other single substrate amino acid position only. Our results show that the combination of at least three particular amino acids is needed in the substrate for a cleavage event to occur. Only by combining and analyzing massive amounts of HIV proteome data it was possible to discover these novel and general patterns of physico-chemical substrate cleavage determinants. Our study is an example how computational biology methods can advance the understanding of the viral interactomes. PMID- 17427232 TI - Classifying the intensity of pediatric cancer treatment protocols: the intensity of treatment rating scale 2.0 (ITR-2). AB - BACKGROUND: To develop and validate a method of classifying the intensity of pediatric oncology treatments using four operationally defined categories of treatment intensity. PROCEDURE: An earlier version of a rating scale, the intensity of treatment rating (ITR), was revised and validated in two phases. In the Scale Construction phase, three criterion raters revised the ITR items based on consensus and item agreement data from pediatric oncologist raters (N = 15). In the Scale Validation phase, the new ITR-2 items were validated using a second set of pediatric oncologists external to our institution (N = 12). In addition, a third group of raters (N = 16), was employed to assess inter-rater reliabilities for 12 patients at varying levels of treatment intensities. RESULTS: Agreement between criterion ratings and the median of external raters for all 34 items on the ITR-2 was very high (r = 0.95, range 0.71-0.91). When the ITR-2 was used to rate 12 patient examples, the inter-rater agreement among pediatric oncologists was also very high (median agreement between criterion-rater pairs: r = 0.87; overall relatedness among 16 raters: r(ICC) = 0.83). CONCLUSIONS: The revised ITR Scale 2.0 (ITR-2) is a valid and reliable scale for classifying the intensity of pediatric oncology treatments. PMID- 17427233 TI - In cyclosporine induced neurotoxicity, is tacrolimus an appropriate substitute or is it out of the frying pan and into the fire? PMID- 17427234 TI - Composite adrenal anaplastic neuroblastoma and virilizing adrenocortical tumor with germline TP53 R248W mutation. AB - Composite tumors are extremely rare. Such tumors in adrenal glands are usually of neuroendocrine-neural type and occur mostly in adults. Their pathogenesis remains elusive. We report a patient with composite neuroblastoma (NB), adrenocortical tumor (ACT), and Li-Fraumeni syndrome (LFS) with germline TP53 R248W mutation. LFS predisposes to the development of leukemia, sarcomas, adrenocortical and breast carcinomas, brain tumors and, questionably, NB. A unique correlation between a single TP53 mutation (R337H) and ACT has been reported in southern Brazilian children. It remains unclear at this time whether a similar association of NB and R248W in patients with LFS exists. PMID- 17427236 TI - Estrous-cycle-dependent hippocampal levels of signaling proteins. AB - There is information that proteins are expressed in a hormone-dependent manner but no systematic study on this subject has been carried out to the best of our knowledge. We therefore decided to investigate protein expression in a well studied brain area, the hippocampus, in female rats at various phases of the estrous cycle and in male rats. Male and female OFA Sprague-Dawley rats were used in the studies and estrous phases were determined using vaginal smears and females were grouped according to PE, E, ME, and DE. Hippocampal tissue was taken, proteins extracted, run on two-dimensional gel electrophoresis and proteins were identified by mass spectrometry methods (MALDI-TOF-TOF and nano-LC ESI-MS/MS). Individual signaling protein levels quantified by specific software were shown to depend on sex and phase of the estrous cycle. These include NG,NG dimethylarginine dimethylaminohydrolase for nitric oxide signaling, stathmin, SH3 domain protein 2A, SH3 domain protein 2B, S100 calcium binding protein B, calcyclin-binding protein, Syndapin I, GTPase HRas, guanine nucleotide-binding proteins, septin 8, G-septin alpha, phosphtidylethanolamine-binding protein, several protein phosphatases. Results from this study, although increasing complexity of protein knowledge, may help to design further investigations at the protein level and may assist to interpret literature on protein expression and brain protein levels. PMID- 17427237 TI - Psoriasis during anti-tumor necrosis factor-alpha therapy for Crohn's disease. PMID- 17427238 TI - Rapid onset of ulcerative colitis after treatment with PEG-interferon plus ribavirin for chronic hepatitis C. PMID- 17427239 TI - Can regular use of 5-ASAs protect against the development of dysplasia and colorectal cancer in patients with ulcerative colitis? PMID- 17427240 TI - Fulminant Crohn's colitis: when only an antibody will do. AB - BACKGROUND: Fulminant Crohn's colitis complicated by toxic dilatation of the large bowel usually requires surgery, which carries a substantial morbidity and mortality rate. METHODS: We report the case of a female with steroid refractory fulminant Crohn's colitis complicated by toxic megacolon who was successfully treated with the antitumor necrosis factor monoclonal antibody adalimumab. RESULTS: Adalimumab induced rapid clinical response and remission, thereby avoiding emergency colectomy. CONCLUSIONS: This is the first report of treatment of fulminant Crohn's colitis with an antitumor necrosis factor antibody and the first report of the use of adalimumab for fulminant colitis of any cause. PMID- 17427241 TI - Epithelial barrier disruption allows nondisease-causing bacteria to initiate and sustain IBD in the IL-10 gene-deficient mouse. AB - BACKGROUND: In the IL-10 gene-deficient mouse model, development of intestinal inflammation is associated with a defect in epithelial barrier integrity that is thought to allow sufficient passage of bacteria or bacterial antigens to initiate a mucosal immune response. Microbial monoassociation experiments into axenic animals have shown that some, but not all, endogenous bacteria will initiate an intestinal inflammatory response. For instance, Bacteroides vulgatus does not initiate intestinal inflammation in axenic IL-10 gene-deficient mice. We investigated whether B. vulgatus requires concomitant disruption of the intestinal epithelial barrier integrity in order to initiate an inflammatory response. METHODS: We first identified a dose of the indomethacin that would cause a primary disruption of the epithelial barrier without causing intestinal inflammation. IL-10 axenic mice were then administered this dose of indomethacin in their drinking water for 7 days and concomitantly monoassociated, by oral gavage, with B. vulgatus. RESULTS: Indomethacin treatment (2 microg/g/d) for 7 days resulted in disruption of epithelial barrier integrity, but it caused neither a systemic inflammatory response nor a mucosal inflammatory response in the colon or cecum. Monoassociation with B. vulgatus alone did not lead to a mucosal inflammatory response, despite a measurable systemic response. In contrast, administration of indomethacin plus B. vulgatus-monoassociation resulted in a marked intestinal inflammatory response in colon and cecum. CONCLUSIONS: Our data show that, in a genetically predisposed animal model, the nondisease-causing endogenous bacteria, B. vulgatus, is able to cause an intestinal inflammatory response provided that disruption of the intestinal epithelial barrier has occurred. PMID- 17427243 TI - It's not all in the genes. PMID- 17427242 TI - Reduced anterior hippocampal formation volume in hyponatremic schizophrenic patients. AB - Diminished hippocampal volume occurs in the anterior segment of some schizophrenic patients, and in the posterior segment in others. The significance of hippocampal pathology in general and these segmental differences in specific is not known. Several lines of evidence suggest anterior hippocampal pathology underlies the life-threatening hyponatremia seen in a subgroup of patients with schizophrenia; therefore our goal was to determine if this region was preferentially diminished in hyponatremic patients. We studied seven polydipsic hyponatremic, ten polydipsic normonatremic, and nine nonpolydipsic normonatremic schizophrenic inpatients, as well as 12 healthy controls. All underwent structural scanning on a high resolution (3.0 T) magnetic resonance imaging (MRI) scanner. Hippocampal formation, amygdala, and third ventricle volumes were manually traced in each subject. The hippocampus was divided at the posterior extent of the uncus, and all structural volumes were corrected for whole brain volume and other significant recognized factors (i.e., age, gender, height, parental education). Despite being overhydrated, anterior hippocampal formation volume was diminished in those with polydipsia and hyponatremia relative to each of the other three groups. Third ventricle volume was larger in this group than in healthy controls but similar to the two patient groups. Posterior hippocampal and amygdala volumes did not differ between groups. Other potential confounds (e.g., water imbalance) either had no effect or accentuated these differences. We conclude the anterior hippocampal formation is smaller in hyponatremic schizophrenic patients, thereby linking an important and objective clinical feature of schizophrenia to a neural pathway that can be investigated in animal models. The findings strengthen the hypothesis that anterior hippocampal formation pathology disrupts functional connectivity with other limbic structures in schizophrenia. PMID- 17427244 TI - Assessment of reliability and validity of IBD phenotyping within the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK) IBD Genetics Consortium (IBDGC). AB - BACKGROUND: The NIDDK IBD Genetics Consortium (IBDGC) collects DNA and phenotypic data from inflammatory bowel disease (IBD) subjects to provide a resource for genetic studies. No previous studies have been performed on the reliability and validity of phenotypic determinations in either Crohn's disease (CD) or ulcerative colitis (UC) using primary records. Our aim was to determine the reliability and validity of these phenotypic assessments. METHODS: The de identified records of 30 IBD patients were reviewed by 2 phenotypers per center using a standard protocol for phenotypic assessment. Each phenotyper evaluated 10 charts on 2 occasions 5 months apart. Reliability was expressed as the kappa (kappa) statistic. Performance characteristics were determined by comparison to a consensus-derived "gold standard" and by generation of receiver operating characteristic (ROC) curves. RESULTS: Agreement for diagnosis was excellent (kappa = 0.82; 95% confidence interval [CI]: 0.71-0.92). Agreement for CD location was good for jejunal, ileal, colorectal, and perianal disease with kappa between 0.60 and 0.74 but was fair for esophagogastroduodenal (kappa = 0.36). Agreement for UC extent (kappa = 0.67; 95% CI: 0.48-0.85), and CD behavior (kappa = 0.67; 95% CI: 0.49-0.83) were very good. Area under the ROC curves was greater than 0.84 for diagnosis, CD behavior, UC extent, and ileal and colonic CD location. CONCLUSIONS: IBD phenotype classification using a standard protocol exhibited very good to excellent inter- and intrarater agreement and validity. This study highlights the importance of standard protocols in generating reliable and valid phenotypic assessments. The data will facilitate estimates of phenotyping misclassification rates that should be considered when making inferences from IBD genotype-phenotype studies. PMID- 17427245 TI - Gender differences in body composition deficits at diagnosis in children and adolescents with Crohn's disease. AB - BACKGROUND: Childhood Crohn's disease (CD) is associated with poor growth and decreased body mass index (BMI); however, lean mass (LM) and fat mass (FM) deficits prior to therapy have not been characterized. OBJECTIVES: To quantify LM and FM in incident pediatric CD subjects and controls, and to identify determinants of LM and FM deficits. METHODS: Whole body LM and FM were assessed using DXA in 78 CD subjects and 669 healthy controls, ages 5-21 yr. Gender specific z-scores for LM (LM-Ht) and FM (FM-Ht) relative to height were derived using log linear regression models in the controls. Multivariate linear regression models adjusted for potential confounders. RESULTS: CD was associated with significantly lower height and BMI for age. Within CD subjects, FM-Ht and LM Ht were significantly lower in females compared with males (FM-Ht z: -0.66+/-0.83 vs. -0.08+/-0.95, p<0.01; LM-Ht z: -1.12+/-1.12 vs. -0.57+/-0.99, p<0.05). In females, CD was associated with significantly lower LM-Ht (p<0.001) and FM-Ht (p=0.001), adjusted for age, race and Tanner stage, compared with controls. LM and FM deficits were significantly greater in older females with CD; 47% of adolescent females had LM-Ht or =3, marked disturbance of consciousness and severe hand paresis. CONCLUSION: We did not find any impact of APOE genotype on mortality or poor outcome 1 year after IS. PMID- 17427279 TI - Hemodynamic cerebral ischemia during carotid endarterectomy evaluated by intraoperative monitoring and post-operative diffusion-weighted imaging. AB - OBJECTIVE: We used the result of monitoring to evaluate patients with post operative neurological deficits attributable to hemodynamic cerebral ischemia owing to cross-clamping of the carotid artery. METHODS: We evaluated 131 carotid endarterectomy (CEA) procedures performed on 118 patients, 96 men and 22 women ranging in age from 38 to 82 years (mean: 67.1 years). For monitoring, we used a combination of somatosensory evoked potential (SEP), functional dynamic electroencephalography (EEG), near-infrared spectroscopy (NIRS) and transcranial Doppler (TCD). Patients who awoke with neurological deficits after CEA immediately underwent diffusion-weighted imaging (DWI) and magnetic resonance angiography (MRA). RESULTS: In 30 of the 131 procedures (22.9%), intraoperative monitoring disclosed abnormalities after cross-clamping of the internal carotid artery (ICA). In two of these 30 patients, shunt was not introduced, because of full recovery of monitoring after blood pressure increasing, however, one patient demonstrated transient ischemic attack (TIA). In six of remaining 28 patients who need shunt, transient hemodynamic cerebral ischemia occurred, however, all patients recovered gradually within 18 hours after CEA. No new lesions were detected on post-operative DWI of the seven patients and MRA demonstrated good patency of the carotid artery. The other 101 patients whose intraoperative monitoring after cross-clamping of the ICA did not disclose abnormalities demonstrated no hemodynamic TIA. CONCLUSION: Hemodynamic ischemia owing to cross clamping of the ICA is rare in patients treated by CEA. However, in patients manifesting neurological deficits upon awakening from CEA, DWI and MRA should be performed immediately to facilitate their prompt treatment. PMID- 17427280 TI - Relative distribution of plasma flow markers and red blood cells across BBB openings in acute cerebral ischemia. AB - Acute blood-brain barrier (BBB) opening in cerebral ischemia is an often observed but seldom studied phenomenon. Increased permeability has been implicated with several consequences including exacerbating ischemic injury, leading to hemorrhagic transformation (HT) and also predictive of chronic damage and a way of delivering therapeutics to the diseased parts of brain. Very few studies have investigated the 'size' of such acute openings. Herein the blood-brain distribution of fluorescent isothiocyanate (FITC)- labeled red blood cells (RBCs; approximately 5 tm in diameter) and two different sized plasma flow markers in cerebral microvessels was studied by laser scanning confocal microscopy (LSCM) 6 and 24 hours after the onset of a 3 hour period of focal ischemia. At hour 6, Evans blue-tagged albumin [EB-Alb; molecular weight (MW)= 68 kDa, Stokes-Einstein radius=37 A], a marker of both plasma flow and BBB opening, was seen both inside and around microvessels whereas the RBCs were only intravascular. FITC-labeled dextran (FITC-dextran; MW=2000 kDa, Stokes-Einstein radius = approximately 150 A), another plasma flow tracer, had not leaked across the BBB into the tissue at this time. At hour 24, both RBCs and FITC-dextran were found extravascularly along with EB-Alb. We postulate that smaller sized openings in BBB at hour 6 limited the leaking of the two large tracers (RBCs and FITC-dextran) and that such size-dependency was lost by 24 hours with the progression of the ischemic injury. PMID- 17427281 TI - Severe intracranial bleedings during endovascular procedures: outcome of surgically treated patients. AB - OBJECTIVE: Severe intracranial bleedings (SIBs) during endovascular procedures (EPs) are accompanied by acute intracranial hypertension and brain herniation signs. The purpose of this study was to determine the effectiveness of urgent surgical management and its related patient outcome in cases with such a fatal complication. METHOD: Medical records were reviewed retrospectively for the last 750 patients treated in our department in the past 12 years with acute non traumatic intracranial bleeding, who underwent a diagnostic or therapeutic EP. Patients with a severe intra-procedural bleeding episode undergoing urgent surgical management (within 30 minutes after bleeding) were analysed. RESULTS: Fourteen of 750 patients with ruptured vascular malformations presented a new SIB during EP. In nine patients, this occurred during initial angiography, two during aneurysm coiling, two during balloon angioplasty and one during arteriovenous malformation (AVM) embolization. The neurological condition 6 months later was good (independents patients) in seven cases with only a mild disability in two of them. Two patients showed a severe disability. Four patients died without recovering their consciousness. One patient presented a satisfactorily course but died weeks later owing to a pulmonary embolism. CONCLUSION: Despite the fatal spontaneous prognosis of severe intracranial bleeding occurring during endovascular diagnostic or therapeutic procedures, a favorable outcome can be expected if an appropriated treatment was set within 30 minutes of the bleeding. PMID- 17427282 TI - Alzheimer's disease and coffee: a quantitative review. AB - PURPOSE: To estimate the pooled risk of coffee consumption for Alzheimer's disease (AD). MATERIAL AND METHODS: We have reviewed all observational studies that evaluated the association between AD risk and coffee consumption. Four studies were identified: two case-control studies and two cohorts. These studies were carried out between 1990 and 2002. RESULTS: There was an obvious protective effect of coffee consumption in the pooled estimate [risk estimate: 0.73 (95% confidence interval: 0.58-0.92)]. However, the homogeneity test was highly significant (p<0.01), indicating heterogeneity across the pooled studies. Pooled analysis applying the random effect model was 0.79 with 95% confidence interval overlapping unity (95% confidence interval: 0.46-1.36). Three studies assessed coffee consumption by interview questionnaire. The risk of AD in coffee consumers versus non-consumers in studies that used interview questionnaire had a pooled risk estimate of 0.70 with 95% confidence interval 0.55-0.90. CONCLUSION: Although our pooled estimates show that coffee consumption is inversely associated with the risk of AD, the four studies had heterogeneous methodologies and results. Further prospective studies evaluating the association between coffee consumption and AD are strongly needed. PMID- 17427283 TI - A marine-derived acidic oligosaccharide sugar chain specifically inhibits neuronal cell injury mediated by beta-amyloid-induced astrocyte activation in vitro. AB - In Alzheimer's disease (AD), beta-amyloid (Abeta) plaques are surrounded by activated astrocytes and microglia. A growing body of evidence suggests that these activated astrocytes contribute to neurotoxicity through the induction of inflammatory cytokines and the production of oxidative stress mediators. Thus, a compound inhibiting Abeta-induced activation of astrocytes may lead to a novel therapy for AD. Our current work investigates the roles of acidic oligosaccharide sugar chain (AOSC), derived from brown algae Echlonia Kurome Okam, on Abeta induced inflammatory responses and cytotoxicity. We observed that AOSC inhibited the toxicity and apoptosis in SH-SY5Y cell line induced by Abeta-stimulated astrocytes conditioned medium. We found that AOSC inhibited the reactive phenotype of astrocytes, blocked cellular oxidative stress, reduced the production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 and prevented the influx of Ca2+. Thus, our results indicate that AOSC might be a potentially therapeutic compound for AD. PMID- 17427284 TI - Activation of the RAGE pathway: a general mechanism in the pathogenesis of polyneuropathies? AB - OBJECTIVES: Binding of ligands to the receptor for advanced glycation end products (RAGE) results in activation of the transcription factor NF-kappaB and subsequent expression of NF-kappaB regulated cytokines and is a possible pathomechanism in diabetic and in vasculitic polyneuropathies (PNP). We wanted to investigate whether the newly discovered RAGE pathway also contributes to the pathogenesis of various other PNP. METHODS: The presence of the RAGE ligand Nepsilon-Carboxymethyllysine (CML), the receptor itself and NF-kappaBp65 was studied in sural nerve biopsies of patients with alcohol-associated PNP (n=5), PNP owing to vitamin B12 deficiency (n=5), chronic inflammatory demyelinating PNP (CIDP, n=10), Charcot-Marie-Tooth disease (CMT) I or II (n= 10), PNP caused by monoclonal gammopathy of unknown significance (MGUS) (n=5), idiopathic PNP (n=10) and five normal controls by immunohistochemistry. Biopsies of either ten patients with diabetic and vasculitic PNP served as positive controls. RESULTS: CML, RAGE and NF-kappaBp65 were found in co-localization in epineurial vessels in PNP owing to vitamin B12 deficiency, diabetes and vasculitis and in the perineurium in diabetic PNP, vasculitic PNP and in some cases in CIDP and vitamin B12 deficiency. Only diabetic subjects demonstrated co-expression of the three antigens in endoneurial vessels. Increased CML, RAGE and NF-kappaBp65 expression was detected in endoneurial and epineurial mononuclear cells in CIDP and in vasculitic PNP. Additionally, RAGE expression in Schwann cells was significantly increased in diabetic PNP. DISCUSSION: These data suggest that activation of the RAGE pathway might contribute to the pathogenesis of CIDP, PNP owing to vitamin B12 deficiency, diabetes and vasculitis, whereas it does not seem to be involved in the pathogenesis of PNP owing to alcohol, MGUS, CMT I or II and idiopathic PNP. PMID- 17427285 TI - Peptide anchoring spore coat assembly to the outer forespore membrane in Bacillus subtilis. AB - Spore formation in Bacillus subtilis involves the formation of a thick, proteinaceous shell or coat that is assembled around a specialized membrane known as the outer forespore membrane. Here we present evidence that the assembling coat is tethered to the outer forespore membrane by a 26-amino-acid peptide called SpoVM, which is believed to form an amphipathic helix. We show that proper localization of SpoVM is dependent on SpolVA, a morphogenetic protein that forms the basement layer of the spore coat, and conversely, that proper localization of SpoIVA is dependent on SpoVM. Genetic, biochemical and cytological evidence indicates that this mutual dependence is mediated in part by contact between an amino acid side-chain located near the extreme C-terminus of SpoIVA and an amino acid side-chain on the hydrophilic face of the SpoVM helix. Evidence is also presented that SpoVM adheres to the outer forespore membrane via hydrophobic, amino acid side-chains on the hydrophobic face of the helix. The results suggest that the SpoVM helix is oriented parallel to the membrane with the hydrophobic face buried in the lipid bilayer. PMID- 17427286 TI - Identification of pseudomurein cell wall binding domains. AB - Methanothermobacter thermautotrophicus is a methanogenic Gram-positive microorganism with a cell wall consisting of pseudomurein. Currently, no information is available on extracellular pseudomurein biology and so far only two prophage pseudomurein autolysins, PeiW and PeiP, have been reported. In this paper we show that PeiW and PeiP contain two different N-terminal pseudomurein cell wall binding domains. This finding was used to identify a novel domain, PB007923, on the M. thermautotrophicus genome present in 10 predicted open reading frames. Three homologues were identified in the Methanosphaera stadtmanae genome. Binding studies of fusion constructs of three separate PB007923 domains to green fluorescent protein revealed that it also constituted a cell wall binding domain. Both prophage domains and the PB007923 domain bound to the cell walls of Methanothermobacter species and fluorescence microscopy showed a preference for the septal region. Domain specificities were revealed by binding studies with other pseudomurein-containing archaea. Localized binding was observed for M. stadtmanae and Methanobrevibacter species, while others stained evenly. The identification of the first pseudomurein cell wall binding domains reveals the dynamics of the pseudomurein cell wall and provides marker proteins to study the extracellular pseudomurein biology of M. thermautotrophicus and of other pseudomurein-containing archaea. PMID- 17427287 TI - Dimeric structure of the cell shape protein MreC and its functional implications. AB - The bacterial actin homologue MreB forms helical filaments in the cytoplasm of rod-shaped bacteria where it helps maintain the shape of the cell. MreB is co transcribed with mreC that encodes a bitopic membrane protein with a major periplasmic domain. Like MreB, MreC is localized in a helical pattern and might be involved in the spatial organization of the peptidoglycan synthesis machinery. Here, we present the structure of the major, periplasmic part of MreC from Listeria monocytogenes at 2.5 A resolution. MreC forms a dimer through an intimate contact along an N-terminal alpha-helix that connects the transmembrane region with two C-terminal beta-domains. The translational relationship between the molecules enables, in principle, filament formation. One of the beta-domains shows structural similarity to the chymotrypsin family of proteins and possesses a highly conserved Thr Ser dipeptide. Unexpectedly, mutagenesis studies show that the dipeptide is dispensable for maintaining cell shape and viability in both Escherichia coil and Bacillus subtilis. Bacterial two-hybrid experiments reveal that MreC Interacts with high-molecular-weight penicillin-binding proteins (PBPs), rather than with low-molecular-weight endo- and carboxypeptidases, indicating that MreC might act as a scaffold to which the murein synthases are recruited in order to spatially organize the synthesis of new cell wall material. Deletion analyses indicate which domains of B. subtilis MreC are required for interaction with MreD as well as with the PBPs. PMID- 17427288 TI - Interaction between FtsW and penicillin-binding protein 3 (PBP3) directs PBP3 to mid-cell, controls cell septation and mediates the formation of a trimeric complex involving FtsZ, FtsW and PBP3 in mycobacteria. AB - In bacteria, biogenesis of cell wall at the division site requires penicillin binding protein 3 (PBP3) (or Ftsl). Using pull-down, bacterial two-hybrid, and peptide-based interaction assays, we provide evidence that FtsW of Mycobacterium tuberculosis (FtsWMTB) interacts with PBP3 through two extracytoplasmic loops. Pro306 in the larger loop and Pro386 in the smaller loop of FtsW are crucial for these interactions. Fluorescence microscopy shows that conditional silencing of ftsW in Mycobacterium smegmatis prevents cell septation and positioning of PBP3 at mid-cell. Pull-down assays and conditional depletion of FtsW in M. smegmatis provide evidence that FtsZ, FtsW and PBP3 of mycobacteria are capable of forming a ternary complex, with FtsW acting as a bridging molecule. Bacterial three hybrid analysis suggests that in M. tuberculosis, the interaction (unique to mycobacteria) of FtsZ with the cytosolic C-tail of FtsW strengthens the interaction of FtsW with PBP3. ftsW of M. smegmatis could be replaced by ftsW of M. tuberculosis. FtsWMTB could support formation of the FtsZ-FtsW-PBP3 ternary complex in M. smegmatis. Our findings raise the possibility that in the genus Mycobacterium binding of FtsZ to the C-tail of FtsW may modulate its interactions with PBP3, thereby potentially regulating septal peptidoglycan biogenesis. PMID- 17427289 TI - SigmaE-dependent small RNAs of Salmonella respond to membrane stress by accelerating global omp mRNA decay. AB - The bacterial envelope stress response (ESR) is triggered by the accumulation of misfolded outer membrane proteins (OMPs) upon envelope damage or excessive OMP synthesis, and is mediated by the alternative sigma factor, sigmaE. Activation of the GE pathway causes a rapid downregulation of major omp mRNAs, which prevents further build-up of unassembled OMPs and liberates the translocation and folding apparatus under conditions that require envelope remodelling. The factors that facilitate the rapid removal of the unusually stable omp mRNAs in the ESR were previously unknown. We report that in Salmonella the ESR relies upon two highly conserved, sigmaE-controlled small non-coding RNAs, RybB and MicA. By using a transcriptomic approach and kinetic analyses of target mRNA decay in vivo, RybB was identified as the factor that selectively accelerates the decay of multiple major omp mRNAs upon induction of the ESR, while MicA is proposed to facilitate rapid decay of the single ompA mRNA. In unstressed bacterial cells, the two oE dependent small RNAs function within a surveillance loop to maintain envelope homeostasis and to achieve autoregulation of oE. PMID- 17427290 TI - Developmental regulation of the glyoxylate cycle in the human pathogen Penicillium marneffei. AB - Penicillium marneffei is a thermally dimorphic opportunistic human pathogen with a saprophytic filamentous hyphal form at 25 degrees C and a pathogenic unicellular yeast form at 37 degrees C. During infection. P. marneffei yeast cells exist intracellularly in macrophages. To cope with nutrient deprivation during the infection process, a number of pathogens employ the glyoxylate cycle to utilize fatty acids as carbon sources. The genes which constitute this pathway have been implicated in pathogenesis. To investigate acetate and fatty acid utilization, the acuD gene encoding a key glyoxylate cycle enzyme (isocitrate lyase) was cloned. The acuD gene is regulated by both carbon source and temperature in P. marneffei, being strongly induced at 37 degrees C even in the presence of a repressing carbon source such as glucose. When introduced into the non-pathogenic monomorphic fungus Aspergillus nidulans, the P. marneffei acuD promoter only responds to carbon source. Similarly, when the A. nidulans acuD promoter is introduced into P. marneffei it only responds to carbon source suggesting that P. marneffei possesses both cis elements and trans-acting factors to control acuD by temperature. The Zn(II)2Cys6 DNA binding motif transcriptional activator FacB was cloned and is responsible for carbon source-, but not temperature-, dependent induction of acuD. The expression of acuD at 37 degrees C is induced by AbaA, a key regulator of morphogenesis in P. marneffei, but deletion of abaA does not completely eliminate temperature-dependent induction, suggesting that acuD and the glyoxylate cycle are regulated by a complex network of factors in P. marneffei which may contribute to its pathogenicity. PMID- 17427291 TI - Hypermutagenesis in mutA cells is mediated by mistranslational corruption of polymerase, and is accompanied by replication fork collapse. AB - Elevated mistranslation induces a mutator response termed translational stress induced mutagenesis (TSM) that is mediated by an unidentified modification of DNA polymerase III. Here we address two questions: (i) does TSM result from direct polymerase corruption, or from an indirect pathway triggered by increased protein turnover? (ii) Why are homologous recombination functions required for the expression of TSM under certain conditions, but not others? We show that replication of bacteriophage T4 in cells expressing the mutA allele of the glyVtRNA gene (Asp-Gly mistranslation), leads to both increased mutagenesis, and to an altered mutational specificity, results that strongly support mistranslational corruption of DNA polymerase. We also show that expression of mutA, which confers a recA-dependent mutator phenotype, leads to increased lambdoid prophage induction (selectable in vivo expression technology assay), suggesting that replication fork collapse occurs more frequently in mutA cells relative to control cells. No such increase in prophage induction is seen in cells expressing alaVGlu tRNA (Glu-->Ala mistranslation), in which the mutator phenotype is recA-independent. We propose that replication fork collapse accompanies episodic hypermutagenic replication cycles in mutA cells, requiring homologous recombination functions for fork recovery, and therefore, for mutation recovery. These findings highlight hitherto under-appreciated links among translation, replication and recombination, and suggest that translational fidelity, which is affected by genetic and environmental signals, is a key modulator of replication fidelity. PMID- 17427292 TI - Hypnosis and the treatment of posttraumatic conditions: an evidence-based approach. AB - This article reviews the evidence for the use of hypnosis in the treatment of posttraumatic conditions including posttraumatic stress disorder and acute stress disorder. The review focuses on empirically supported principles and practices and suggests that hypnosis can be a useful adjunctive procedure in the treatment of posttraumatic conditions. Cognitive-behavioral and exposure-based interventions, which have the greatest empirical support, are highlighted, and an illustrative case study is presented. PMID- 17427293 TI - Evidence-based hypnotherapy for asthma: a critical review. AB - Asthma is a chronic disease with intermittent acute exacerbations, characterized by obstructed airways, hyper-responsiveness, and sometimes by chronic airway inflammation. Critically reviewing evidence primarily from controlled outcome studies on hypnosis for asthma shows that hypnosis is possibly efficacious for treatment of symptom severity and illness-related behaviors and is efficacious for managing emotional states that exacerbate airway obstruction. Hypnosis is also possibly efficacious for decreasing airway obstruction and stabilizing airway hyper-responsiveness in some individuals, but there is insufficient evidence that hypnosis affects asthma's inflammatory process. Promising research needs to be replicated with larger samples and better designs with careful attention paid to the types of hypnotic suggestions given. The critical issue is not so much whether it is used but how it is used. Future outcome research must address the relative contribution of expectancies, hypnotizability, hypnotic induction, and specific suggestions. PMID- 17427294 TI - [Academic freedom and value-free science: Max Weber and the university teacher's congress at Jena, 1908]. PMID- 17427295 TI - [Missing in World War II: the development of the tracing service of the German Red Cross]. PMID- 17427296 TI - [History as palimpsest: writing and reading on the "children of the perpetrators"]. PMID- 17427297 TI - Reader responds to "The epidemic of chronic kidney disease". PMID- 17427298 TI - Private law enforcement in Norwegian history: the husband's right to chastise his wife. PMID- 17427299 TI - Readers' response to "Time to reform physician relicensure". PMID- 17427300 TI - [The social engineer in the "Interest Office": a study in Swedish welfare history]. PMID- 17427301 TI - Reader's response to "Endocannabinoids--the brain's own marijuana--may be linked to the metabolic syndrome". PMID- 17427302 TI - Reader's response and author's reply to "Our patients are sicker--posting grades on the classroom, office, or hospital door". PMID- 17427303 TI - [The dream that came true: state-subsidized vacations for needy housewives]. PMID- 17427304 TI - Readers' responses and author's reply to "The reality of 'traditional Chinese' medicines". PMID- 17427305 TI - Diagnosis: paraspinal (retropharyngeal) abscess with cervical osteomyelitis. PMID- 17427306 TI - Diagnosis: cystic fibrosis. PMID- 17427307 TI - When to divulge patient information. PMID- 17427308 TI - Reader's response and author's reply to "Hope in the neonatal intensive care nursery". PMID- 17427309 TI - Readers' response and author's reply to "Laboratory results that should be ignored". PMID- 17427310 TI - Reader's response to "The future of emergency care in the United States". PMID- 17427311 TI - Joint replacement in the overweight patient - a view from the Patient Liaison Group. PMID- 17427312 TI - 'Autistic regression associated with seizure onset in an infant with tuberous sclerosis'. PMID- 17427313 TI - [Quality, efficiency, and skill: the practical use of science in Swedish 18th century political decisionmaking]. PMID- 17427314 TI - The institution of retirement on Scanian estates in the nineteenth century. PMID- 17427315 TI - [The Swedish labor movement's attitude toward the cultivation of the aesthetic taste of the masses]. PMID- 17427316 TI - ["The interest of humanity in general and my sex in particular": the letters of Julie Bondeli on upbringing and the education of women]. PMID- 17427318 TI - An up side to ozone depletion? PMID- 17427317 TI - Public statement: guidelines for the assessment and management of pain in rodents and rabbits. AB - The ACLAM Analgesia Task Force was appointed by ACLAM President Diane Gaertner in 2003. The charge to the Task Force was to develop guidelines that could be used by veterinarians, scientists and IACUCs in helping to provide appropriate assessment and management of pain in rodents, with the understanding that ultimately the clinical veterinarian on site at the institution must make decisions relevant to a specific animal or animals and/or protocol. The guidelines were not to be developed as, and should not be used as, requirements. To complete its charge the Task Force reviewed and cited, in a comprehensive manner, available data-based literature in writing the paper. In the course of completion of the document, ACLAM Board of Directors (BOD) reviewed an early draft and at that time also appointed 3 ACLAM diplomates with particular expertise in assessment and management of pain in rodents to act as reviewers of the draft. The Task Force members responded to the critiques and comments submitted by both the BOD and the 3 reviewers. This revised draft was then placed on the ACLAM website for comments from the entire College. The draft was well received by responding diplomates. Comments from the membership were reviewed and discussed by the Task Force, and most were incorporated into the final draft manuscript. The draft then received final review and editing by the ACLAM Publications Committee Chairman, and was accepted in the format presented here. Despite this extensive vetting process through ACLAM, readers should nonetheless be aware that because this document represents the approved statement of an AALAS affiliate organization, it has not undergone the usual JAALAS peer review process. I would like to acknowledge the efforts of the Task Force and the leadership of ACLAM for supporting this comprehensive and informative synthesis. The document should serve as a resource to the research community for years to come. PMID- 17427319 TI - Theoretically speaking. Interview by Hilary Crichton. PMID- 17427320 TI - Delivering the goods. PMID- 17427321 TI - Are tracheal dilators useful in stomal stenosis? PMID- 17427322 TI - Medication management with panic disorder in the exercise state: some prescribing guidelines. PMID- 17427323 TI - PEC proposals promise equality for clinicians and managers. PMID- 17427324 TI - Dentistry shortfall adds to PCT woes. PMID- 17427325 TI - GPs' commissioning saves hospital. PMID- 17427326 TI - Strategist slams reform protests. PMID- 17427327 TI - Restructuring. How Middle England turned into a nation of reconfiguration rebels. PMID- 17427328 TI - On winning hearts and minds. PMID- 17427329 TI - The age of uncertainty. Interview by Nick Edwards. PMID- 17427330 TI - Data briefing. Why are admissions on the increase? PMID- 17427331 TI - Mental health. 'People who really want to do this make a difference'. AB - Birmingham's early intervention in psychosis service is the type the government would like to see across England, but roll-out can at best be described as patchy. Models of provision vary widely--from a standalone service to one fully integrated with other youth provision in the area. Research suggests early intervention could save as much as 53 percent on the cost of a conventional service. PMID- 17427332 TI - Health inequality. In the balance. PMID- 17427333 TI - Employ health trainers. PMID- 17427334 TI - Elderly care. Controlling interest. PMID- 17427335 TI - Assisted living nurses monitor chronic diseases. PMID- 17427336 TI - 2003: a year of transition. PMID- 17427337 TI - Conditions improve in 2003, but challenges lie ahead. PMID- 17427338 TI - CMS retools quality measures. PMID- 17427339 TI - Eden alternative with a twist. PMID- 17427340 TI - Gene therapy methods in bone and joint disorders. Evaluation of the adeno associated virus vector in experimental models of articular cartilage disorders, periprosthetic osteolysis and bone healing. AB - BACKGROUND: Gene therapy is a technique that draws on the introduction of new genes into cells for the purpose of treating disease by restoring or adding gene expression. Numerous growth factors and other proteins with the ability to promote the regeneration of tissues in the locomotive system have been identified, but their clinical use is often hindered by delivery problems. In principle, these problems can be overcome by delivering the relevant genes, as the therapeutic substances thereby can be persistently produced directly by local cells at the site of diseases. HEALING OF ARTICULAR CARTILAGE: Articular chondrocytes are receptive to transduction using various gene delivery methods. Following genetic modification, they are capable of sustained expression of transgene products at biologically relevant levels. Our research has proved the AAV vector to be an effective tool for gene delivery to articular chondrocytes in vitro as well as in vivo. To this end, we have demonstrated that the AAV vector mediated TGFbeta1-overexpression stimulates cartilage anabolism. WEAR DEBRIS INDUCED OSTEOLYSIS: The RANKL system may be a key therapeutic target in treatment of aseptic periprosthetic loosening. We investigated whether gene transfer of OPG using an AAV vector has protective effects against orthopaedic wear debris induced bone loss. In osteoclastogenesis and in bone wafer resorption assays, the bioactivity of the transgene OPG was proven by depletion of osteoclastogenesis and reduced bone resorption. Using an in vivo model of debris-induced bone resorption, we demonstrated complete inhibition of osteolysis in animals receiving AAV-OPG gene therapy. FRACTURE HEALING IN RELATION TO OSTEOPOROSIS: The success of future OPG treatment of osteoporosis is highly dependent on its effects on fracture healing and remodelling. Using an in vivo fracture healing model, our studies demonstrated that AAV-OPG gene therapy did not conflict with normal bone healing, in contrast to high-dosage intravenous treatment with OPG. However, AAV-OPG therapy depressed remodelling and integration of the genuine cortical bone at the fracture line. STRUCTURAL BONE ALLOGRAFT HEALING: Structural bone allografts often fracture due to their lack of osteogenic and remodelling potiential. To overcome these limitations, we utilized allografts coated with AAV caALK2 vector that mediated in vivo gene transfer. We showed that the AAV vector was capable of transducing adjacent inflammatory cells and osteoblasts in the fracture callus and that BMP signals delivered via AAV-caALK2 coating induced bone formation directly on the cortical surface of the allograft. CONCLUSION: The presented research may be seen as initial steps towards development of gene therapeutic treatment options for complex orthopaedic diseases. As such, our studies represent proof-of-principle that the rAAV vector promotes efficient gene transfer in vitro to a spectrum of cells with orthopaedic relevance, and that in vivo targeting of somatic tissue with a single administration of a rAAV vector at the time of surgery could be sufficient for long-term expression of therapeutic proteins. Essential to the future success of transgene delivery by rAAV vectors is the absence of an immune response to either the vector or the gene product. Furthermore, development of rAAV vectors with regulatory gene expression needs further attention in future research. PMID- 17427341 TI - Global construction health and safety--what works, what does not, and why? AB - The marginalization of construction workers makes them especially vulnerable in terms of poor occupational health and safety standards and lax enforcement, with resulting disabilities. A sound research base is needed to rectify this situation. Efforts to raise awareness of construction hazards and ameliorate them are described. PMID- 17427342 TI - Building safety--an international perspective. AB - International health and safety standards to protect construction workers are already in effect but are often ignored by management. Conforming to these standards and implementing other measures described by the author could substantially reduce the high incidences of injuries, illnesses, and fatalities arising from work in this industry. PMID- 17427343 TI - Significant developments in occupational health and safety in Australia's construction industry. AB - Construction is Australia's third most dangerous industry. On average, 49 building and construction workers have been killed at work each year since 1997 1998. Securing safer construction workplaces is jeopardized by an Australian government bent on removing union influence. Workers must prove their individual health and safety is at risk or face fines of up to $22,000 over work stoppages. The Australian Chamber of Commerce and Industry view is that occupational health and safety (OHS) legislation is overly complex and biased unfairly against the employer. Significant advances in OHS achieved by construction unions working together with employers and government authorities are now at risk, to the detriment of workers. PMID- 17427344 TI - Struggles for health and safety in the Danish construction industry. AB - An encouraging trend of reductions in accidents and fatalities in the Danish construction industry, brought about by the combined sustained efforts of unions, management, and government, is suffering a reverse. While some large construction companies have achieved excellent safety records through effective internal programs combining rewards and penalties as incentives, the overall picture is worsening as government eases pressures on small and medium-sized enterprises by relaxing occupational health and safety regulations. PMID- 17427345 TI - Protest and survive, the U.K. Construction Safety Campaign: a case study of workers' and victims' family action and legal and regulatory inaction on construction health and safety. AB - The U.K. Construction Safety Campaign, based on grass-roots rank-and-file trade union activity, has both a domestic agenda and a wider sphere with regard to international developments. Using organized protests and media exposure to achieve its ends, the Campaign has contributed substantially to reducing injuries and fatalities in the U.K. construction industry, as well as achieving some justice for victims of the industry's neglect of safe management practices. PMID- 17427346 TI - Cancer and construction: what occupational histories in a Canadian community reveal. AB - From 2000 to 2002, male patients at a Canadian cancer treatment center with new incident head-and-neck or esophageal cancers were invited to participate in a population-based study. The study population included 87 cases and 172 controls. A lifetime-history questionnaire was administered. Odds ratios (ORs) were calculated for occupational groups with a minimum of five cases, adjusted for duration of employment, age, smoking, alcohol, education, and income. A significantly increased risk was shown for construction workers (OR = 2.20; 95% CI 1.25-3.91). This investigation of a set of rare cancers over a limited time period demonstrates the feasibility of this research approach. The increased risk among construction workers supports the need for more comprehensive study of exposures in this occupational group. PMID- 17427347 TI - Is the apparent decrease in injury and illness rates in construction the result of changes in reporting? AB - Injury rates in all industries and in construction in particular have been declining. Inconsistencies in the information suggest some of the apparent decrease may be due to changes in the ways injuries are treated, misclassification of employees, or underreporting. Lost-time injury rates for the largest construction employers declined by as much as 92% between 1988 and 1999. Yet the rate for cases with restricted work activity actually increased from 0.7 to 1.2 per 100 full-time workers between 1990 and 2000, and fatalities among construction workers remain high. In Massachusetts, at least 14% of construction employers misclassified workers as independent contractors, with the effect that injuries to these workers are not recordable. Studies that compare OSHA logs with other data sources find that the OSHA logs do not include a significant proportion of injuries and illnesses identified elsewhere. PMID- 17427348 TI - A roadmap to diffuse ergonomic innovations in the construction industry: there is nothing so practical as a good theory. AB - Despite the availability of resources and practices that would reduce work related morbidity and mortality in the construction industry, their diffusion to workers has been slow, partly because the ties between management and trade workers are weak. In promoting an innovation, it is necessary to target the stakeholders who will be making the decisions related to it. The authors' focus is on ergonomics, but their observations may be applied more broadly to other areas of intervention-effectiveness research. PMID- 17427349 TI - Trainer evaluation of a union-based ten-hour safety and health hazard-awareness program for U.S. construction workers. AB - A web-based survey of union-based outreach instructors evaluated training materials developed to teach OSHA ten-hour hazard-awareness courses to members of the construction trades. Respondents taught an average of five ten-hour hazard awareness courses per year. When asked about hazards commonly encountered by their trainees, 83% identified falls from ladders, with a range of 1-22 hazards identified. Over one third of the trainers taught individuals whose primary languages were not English. Increased interaction with trainees through subsequent phone calls may be a marker of differential training impact. PMID- 17427350 TI - The role of asbestos fiber dimensions in the prevention of mesothelioma. AB - A recent interpretation of the pathogenetic role of asbestos fiber size in the development of mesothelioma and in the possibility of mesothelioma prevention needs clarification. This point of view is based on a biased interpretation of the literature. Epidemiologic, experimental, and molecular evidence suggests that the arguments for the role of fiber size relative to dose, dose-response effect, and genetic susceptibility are scientifically unsound. Their proponent also states that means available in the past for the implementation of dust-control measures and/or personal protective equipment would not have contributed to reducing the frequency of mesothelioma among exposed subjects, an argument again based on invalid assumptions. PMID- 17427351 TI - The Rochdale asbestos cancer studies and the politics of epidemiology: what you see depends on where you sit. AB - The history of the exploitation of epidemiology by the U.K. asbestos industry and the subsequent obscuring of the disastrous results of exposures is presented, exploring in particular the roles of Sir Richard Doll and his colleagues. Epidemiology, often regarded as a neutral science, is susceptible to socio political influences. PMID- 17427352 TI - Ban on asbestos diaphragms in the chlorine-related chemical industry and efforts toward a worldwide ban. AB - The EU Directive 1999/77/EC, prohibiting new applications of chrysotile, exempted diaphragms used in electrolysis to produce chorine and caustic soda in existing chlor-alkali plants until they reach the end of their service life, or until suitable asbestos-free substitutes become available, whichever is sooner. There is no technical justification for extending this exemption after January 2008, when it is scheduled for review. Economic interests should not take precedence over the social and environmental costs imposed by the asbestos production required for this industry. The EU Scientific Committee on Toxicity, Ecotoxicity and the Environment should not further delay the necessary decision on a total and immediate worldwide ban of the mining, manufacturing, and all uses of asbestos. PMID- 17427353 TI - Popcorn-worker lung caused by corporate and regulatory negligence: an avoidable tragedy. AB - Diacetyl-containing butter flavor was identified as the cause of an outbreak of bronchiolitis obliterans (BO) and other lung diseases in popcorn-plant workers. Litigation documents show that the outbreak was both predictable and preventable. The industry trade organization was aware of BO cases in workers at butter flavoring and popcorn-manufacturing plants but often failed to implement industrial hygiene improvements and actively hid pertinent warning information. Due to weaknesses in the organization and mandates of regulatory bodies, organizations such as NIOSH, OSHA, the FDA, particularly the "generally recognized as safe" (GRAS) system, and the EPA failed to detect and prevent the outbreak, which highlights the need for systemic changes in food-product regulation, including the need for corporations to act responsibly, for stronger regulations with active enforcement, for a restructuring of the GRAS system, and for criminal penalties against corporations and professionals who knowingly hide information relevant to worker protection. PMID- 17427354 TI - A successful workplace program for voluntary counseling and testing and treatment of HIV/AIDS at Heineken, Rwanda. AB - Heineken Breweries launched a workplace HIV/AIDS program at its Rwanda subsidiary in September 2001. By January 25, 2005, 736/2,595 eligible individuals had reported for counseling and HIV testing: 380/521 employees (72.9%), 254/412 spouses (61.7%), 99/1,517 children (6.5%), and 3/145 retired (2.0%). As a result, 109 HIV+ individuals were identified: 62 employees, 34 spouses, 12 children, and 1 retired. In September 2003 an anonymous HIV seroprevalence survey was performed with participation rates of 69.4% for employees, 58.2% for spouses, and 79.7% for adolescents. Using the survey result, the expected number of HIV+ employees was 71, which implies a program uptake of 87.1% (62/71) in this group. Of the identified 109 HIV+ beneficiaries, 42 were on highly active antiretroviral treatment (HAART). In November 2003 a qualitative study of awareness and health seeking behavior of the Heineken Rwanda beneficiaries identified key principles contributing to the success of this program. PMID- 17427355 TI - Industry influence on occupational and environmental public health. AB - Traditional covert influence of industry on occupational and environmental health (OEH) policies has turned brazenly overt in the last several years. More than ever before the OEH community is witnessing the perverse influence and increasing control by industry interests. Government has failed to support independent, public health-oriented practitioners and their organizations, instead joining many corporate endeavors to discourage efforts to protect the health of workers and the community. Scientists and clinicians must unite scientifically, politically, and practically for the betterment of public health and common good. Working together is the only way public health professionals can withstand the power and pressure of industry. Until public health is removed from politics and the influence of corporate money, real progress will be difficult to achieve and past achievements will be lost. PMID- 17427356 TI - Against anti-health epidemiology: corporate obstruction of public health via manipulation of epidemiology. AB - In response to several articles on corporate corruption of science that appeared earlier in this journal, a critic outlined an epistemological model based on an unsupported assertion that epidemiologic evidence is always required to support cause-effect relationships. This model, if adopted, would eliminate compensation to victims of toxic exposures and impede regulation of accepted hazards. Epidemiology is only one element in support of cause-effect determinations. The critic's proposal of an anti-health epidemiology was initially developed by corporations with the goal of providing defense in litigation, and is based not on science but on a corporate need to enhance profits at the expense of public health. PMID- 17427357 TI - "Gulf war syndrome" may be related to circadian dysrhythmia. AB - An Institute of Medicine (IOM) review found that the data obtained from research addressing the health issues of Gulf War veterans do not satisfactorily clarify the origins, extent, and long-term implications of their health problems. The IOM committee concluded that there should have been more screening and medical examinations of deployed personnel before and after service in the Gulf. The many possible causes of the "Gulf War syndrome" examined, however, did not include circadian dysrhythmia or desynchronosis. It would have been possible to determine the level of desynchronosis in the returning Gulf War veterans, and to follow them into their subsequent pursuits to determine whether chronic desynchronosis was present in those who had persistent symptoms. If circadian dysrhythmia is found to be present in veterans now returning from the Gulf, they should receive treatment to correct the problem before they develop chronic desynchronosis. PMID- 17427358 TI - Integrity of scientific evaluations of government agencies. PMID- 17427359 TI - More about differential peeky bias. PMID- 17427360 TI - FIOH-sponsored newsletter. Misrepresentation of asbestos hazards in Zimbabwe. Lack of objectivity and coherence in articulating the subject of chrysotile asbestos in the Bailer et al. article. PMID- 17427361 TI - FIOH-sponsored Newsletter. Misrepresentation of asbestos hazards in Zimbabwe. Zimbabwe can use chrysotile fiber safely and responsibly. PMID- 17427362 TI - On-site clinics are for employers, not workers. PMID- 17427363 TI - Understanding quality in health care. PMID- 17427364 TI - Time to stop whinging about portfolios. PMID- 17427365 TI - Congenital heart disease study day. PMID- 17427366 TI - Ka pai, te roro hiko. PMID- 17427367 TI - Judge expresses 'considerable sympathy' for NAs. PMID- 17427368 TI - Meeting the needs of patients in the last days of life. PMID- 17427369 TI - Dying with dignity in aged care. PMID- 17427370 TI - Helping families get past the missing socks. PMID- 17427371 TI - Caring for cultural differences. PMID- 17427372 TI - Nurses must claim ownership of PDRPS. PMID- 17427373 TI - Understanding funding in aged care. PMID- 17427374 TI - Getting the ear of the minister. PMID- 17427375 TI - The role of an individual case organiser. PMID- 17427376 TI - NZNO seeks changes to the student loan scheme. PMID- 17427377 TI - New red blood cell antigens in dogs and cats--a welcome discovery. PMID- 17427378 TI - Breed associations for canine exocrine pancreatic insufficiency. AB - BACKGROUND: Knowledge of breed associations is valuable to clinicians and researchers investigating diseases with a genetic basis. HYPOTHESIS: Among symptomatic dogs tested for exocrine pancreatic insufficiency (EPI) by canine trypsin-like immunoreactivity (cTLI) assay, EPI is common in certain breeds and rare in others. Some breeds may be overrepresented or underrepresented in the population of dogs with EPI. Pathogenesis of EPI may be different among breeds. ANIMALS: Client-owned dogs with clinical signs, tested for EPI by radioimmunoassay of serum cTLI, were used. METHODS: In this retrospective study, results of 13,069 cTLI assays were reviewed. RESULTS: An association with EPI was found in Chows, Cavalier King Charles Spaniels (CKCS), Rough-Coated Collies (RCC), and German Shepherd Dogs (GSD) (all P < .001). Chows (median, 16 months) were younger at diagnosis than CKCS (median, 72 months, P < .001), but not significantly different from GSD (median, 36 months, P = .10) or RCC (median, 36 months, P = .16). GSD (P < .001) and RCC (P = .015) were younger at diagnosis than CKCS. Boxers (P < .001), Golden Retrievers (P < .001), Labrador Retrievers (P < .001), Rottweilers (P = .022), and Weimaraners (P = .002) were underrepresented in the population with EPI. CONCLUSIONS AND CLINICAL IMPLICATIONS: An association with EPI in Chows has not previously been reported. In breeds with early-onset EPI, immune-mediated mechanisms are possible or the disease may be congenital. When EPI manifests later, as in CKCS, pathogenesis is likely different (eg, secondary to chronic pancreatitis). Underrepresentation of certain breeds among dogs with EPI has not previously been recognized and may imply the existence of breed-specific mechanisms that protect pancreatic tissue from injury. PMID- 17427379 TI - Transient hyperammonemia due to urea cycle enzyme deficiency in Irish wolfhounds. AB - BACKGROUND: Irish Wolfhounds frequently have a congenital portosystemic shunt, but a considerable proportion of the 6- to 8-wk-old pups has hyperammonemia in the absence of portosystemic shunting. This hyperammonemia causes no signs and is transient, normalizing at the age of 3-4 months. HYPOTHESIS: Transient hyperammonemia has a metabolic basis in Irish Wolfhounds. ANIMALS: Two related (same sire) litters of Irish Wolfhounds (17 pups) and their parents were studied. METHODS: Integrity of the portal circulation was examined by ultrasonography and scintigraphy. Absence of parenchymal liver disease was verified by liver biopsy. Amino acid profiles were measured in 4 pups and repeated in 2 of these pups when ammonia concentrations had normalized. The amino acid profiles were compared with those of healthy Irish Wolfhound pups. RESULTS: Fasting venous ammonia concentrations were high (113-622 microg/dL, 65-345 micromol/L) in all pups, whereas bile acids were within reference range in all but 1. The ammonia and bile acid concentrations from all parents were within reference range. Portosystemic shunting was excluded in all but 1 pup. Liver biopsy excluded significant lesions in all 10 pups examined. Hypercitrullinemia was found and persisted even when ammonia had normalized, at the expense of an increase in glutamine and asparagine. CONCLUSIONS AND CLINICAL IMPORTANCE: Citrulline concentrations are controlled by the urea cycle enzymes argininosuccinase and argininosuccinate synthetase, and a defect in either of these enzymes may be responsible for the transient hyperammonemia in Irish Wolfhounds. Resolution of the hyperammonemia is associated with increased activity of alternative metabolic pathways forming glutamine and asparagine. Confirmation requires measurement of enzyme activities in liver tissue. PMID- 17427380 TI - Flexible bronchoscopy and bronchoalveolar lavage in 68 cats (2001-2006). AB - BACKGROUND: Bronchoscopy is an important tool for identifying an underlying etiology for respiratory disease in cats. However, the procedure is challenging, because feline airways are small and prone to bronchoconstriction. HYPOTHESIS: Bronchoscopy and bronchoalveolar lavage (BAL) are appropriate and safe diagnostic procedures in the cat. ANIMALS: Sixty-eight cats. METHODS: Flexible bronchoscopy was performed in all cats with the cats under propofol infusion with jet ventilation. The procedures were reviewed for BAL volumes instilled and recovered and for the number and type of complications with the use of 3 flexible endoscopes < 5.0-mm outer diameter. The BAL procedure was compared among scopes by using a one-way analysis of variance. Complication rates were compared by using chi-square analysis. Significance was set at P < .05. RESULTS: Clinical diagnoses included inflammatory airway disease in 46 of 68 cats, pneumonia in 10 of 68, neoplastic disease in 8 of 68, and other conditions in 4 of 68 cats. Mean lavage volumes instilled for the 3 scopes were 2.62-5.05 mL/kg (range, 0.77-9.38 mL/kg). Mean percent fluid recovered for the 3 scopes was 51-73%, (range, 0 140%). BAL cell counts were adequate for cytologic assessment (> 300 cells/microL) in 61 of 64 cats (97%), and in 107 of 120 samples (89%) collected. Complications occurred in 38% of procedures; however, these were mild in 24% of cats; 6% of cats died or were euthanized after the procedure. Complications were not associated with fluid volume instilled or recovered, and could not be related to the underlying disease process. CONCLUSIONS AND CLINICAL IMPORTANCE: Flexible bronchoscopy with BAL was well tolerated in most cats examined. PMID- 17427381 TI - Relative adrenal insufficiency in dogs with sepsis. AB - BACKGROUND: A syndrome of relative adrenal insufficiency has been identified in septic humans, and is associated with hypotension and death. Relative adrenal insufficiency is generally associated with basal serum cortisol concentration within or above the reference range and a blunted cortisol response to adrenocorticotropic hormone administration. It is unknown whether relative adrenal insufficiency occurs in septic dogs. HYPOTHESIS: That relative adrenal insufficiency occurs in septic dogs, and that relative adrenal insufficiency is associated with hypotension and mortality. ANIMALS: Thirty-three septic dogs admitted to a small animal intensive care unit. METHODS: Dogs were included in the study if they had a known or suspected infectious disease and had systemic inflammatory response syndrome. Dogs were excluded if they had disease or medication history expected to affect the hypothalamic-pituitary-adrenal axis. Serum cortisol and endogenous plasma adrenocorticotropic hormone concentrations were measured before, and serum cortisol concentration measured 1 hour after, intramuscular administration of 250 microg of cosyntropin/dog. The change in cortisol concentration (delta-cortisol) before and after cosyntropin administration was determined in each dog. RESULTS: Hypotension was associated with lower delta-cortisol values (OR 1.3; CI 1.0-1.9; P = .029). delta-Cortisol cutoff of 3.0 microg/dL was most accurate for predicting hypotension, survival to discharge, and 28-day survival. The rate of death in dogs with delta-cortisol < or = 3 microg/dL was 4.1 times that of dogs with delta-cortisol > 3 microg/dL (RR 4.1; CI 1.5-12.3; P = .01). CONCLUSIONS AND CLINICAL RELEVANCE: Delta-cortisol < or = 3 microg/dL after adrenocorticotropic hormone administration is associated with systemic hypotension and decreased survival in septic dogs. PMID- 17427382 TI - Tissue Doppler imaging in Maine Coon cats with a mutation of myosin binding protein C with or without hypertrophy. AB - BACKGROUND: The cardiac myosin binding protein C gene is mutated in Maine Coon (MC) cats with familial hypertrophic cardiomyopathy. HYPOTHESES: Early diastolic mitral annular velocity is incrementally reduced from normal cats to MC cats with only an abnormal genotype to MC cats with abnormal genotype and hypertrophy. ANIMALS: Group 1 consisted of 6 normal domestic shorthair cats, group 2 of 6 MC cats with abnormal genotype but no hypertrophy, and group 3 of 15 MC cats with hypertrophy and abnormal genotype. METHODS: The genotype and echocardiographic phenotype of cats were determined, and the cats were divided into the 3 groups. Tissue Doppler imaging (TDI) of the lateral mitral annulus from the left apical 4 chamber view was performed. Five nonconsecutive measurements of early diastolic mitral annular velocity (EM) or summated early and late diastolic velocity (EAsum) and heart rate were averaged. RESULTS: There was an ordered reduction in Em-EAsum as group number increased (group 1, range 9.7-14.7 cm/s; group 2, range 7.5-13.2 cm/s; group 3, range 4.5-14.1 cm/s; P = .001). Using the lower prediction limit for normal Em-EAsum, the proportion of cats with normal Em-EAsum decreased as the group number increased (P = .001). However, Em-EAsum was reduced in only 3 of 6 cats in group 2. CONCLUSION: The incremental reduction of Em-EAsum as group severity increased indicates that diastolic dysfunction is an early abnormality that occurs before hypertrophy development. TDI measurement of Em or EAsum of the lateral mitral annulus is an insensitive screening test for identification of phenotypically normal, genotypically affected cats. PMID- 17427383 TI - Distinguishing cardiac and noncardiac dyspnea in 48 dogs using plasma atrial natriuretic factor, B-type natriuretic factor, endothelin, and cardiac troponin I. AB - BACKGROUND: It is challenging to differentiate congestive heart failure (CHF) from noncardiac cause of dyspnea. HYPOTHESIS: Circulating concentrations of atrial natriuretic peptide (NT-proANP), B-type natriuretic peptide (BNP), endothelin-I (ET-1), and cardiac troponin-I (cTnI) can be used to help distinguish between cardiac and noncardiac causes of dyspnea in dogs. ANIMALS: Forty-eight client-owned dogs admitted to a veterinary teaching hospital for respiratory distress. METHODS: Blood samples from patients were prospectively obtained. The etiology of dyspnea was determined by using physical examination, thoracic radiographs, and echocardiography. RESULTS: CHF was diagnosed in 22 dogs, and dyspnea of noncardiac origin (noHD group) was diagnosed in 26 dogs. Analyses revealed significant difference between groups for NT-proANP (geometric mean, 95% confidence [CI]; no HD: 0.26 nmol/mL, 95% CI 0.17-1.09; CHF: 1.38 nmol/mL, 95% CI 1.09-1.74 nmol/mL; P < .0001), BNP (noHD: 12.18 pg/mL, 95% CI 10.91-16.17 pg/mL; CHF: 34.97 pg/mL, 95% CI 23.51-52.02 pg/mL; P < .0001), and ET 1 (noHD: 0.32 fmol/mL, 95% CI 0.23-0.46 fmol/mL; CHF: 1.26 fmol/mL, 95% CI 0.83 1.91 fmol/mL; P < .0001). Plasma cTnI concentrations were not significantly different between groups (noHD: 0.29 ng/mL, 95% CI 0.12-0.72 ng/mL; CHF: 0.42 ng/mL, 95% CI 0.18-0.97, P = .53). Receiver operating curves indicated areas under the curve for NT-proANP, BNP, and ET-1 of 0.946, 0.886, and 0.849, respectively. CONCLUSIONS AND CLINICAL IMPORTANCE: Plasma NT-proANP, BNP, and ET 1, but not cTnI, appear useful for distinguishing between dogs with cardiac and noncardiac causes of dyspnea, with plasma NT-proANP having the highest sensitivity (95.5%) and specificity (84.6%). PMID- 17427384 TI - Prospective clinical evaluation of an ELISA B-type natriuretic peptide assay in the diagnosis of congestive heart failure in dogs presenting with cough or dyspnea. AB - BACKGROUND: B-type natriuretic peptide (BNP) is increased in dogs with congestive heart failure (CHF). HYPOTHESIS: The purpose of this study was to evaluate the clinical utility of a novel canine-specific enzyme-linked immunosorbent assay of BNP for the diagnosis of CHF in dogs presenting with either cough or dyspnea. ANIMALS: Three hundred and thirty dogs from 2 large university teaching hospitals. METHODS: We prospectively measured plasma BNP concentrations in 3 groups of dogs: (1) normal adult dogs (n = 75), (2) dogs with asymptomatic heart disease (n = 76), and (3) dogs with cough or dyspnea (n = 179). The final diagnosis of dogs with cough or dyspnea and the severity of CHF (International Small Animal Cardiac Health Council Heart Failure Classification [ISACHC]) were determined by medical record review by a study cardiologist who was blinded to the results of the BNP assay. RESULTS: Dogs with CHF had a higher median BNP concentration (24.6 pg/mL) than dogs with noncardiac causes of cough or dyspnea (2.6 pg/mL) (P < .0001). The area under the curve was 0.91 for the receiver operating curve analysis of the diagnostic accuracy of the BNP measurement to differentiate CHF from other causes of cough or dyspnea. The median BNP concentrations in dogs were 3.0 pg/mL with ISACHC I, 17.8 pg/mL with ISACHC II, and 30.5 pg/mL with ISACHC III. (P < .0001) CONCLUSION AND CLINICAL IMPORTANCE: Measurement of BNP is useful in establishing or in excluding the diagnosis of CHF in dogs with cough or dyspnea. B-type natriuretic peptide concentrations rose significantly as a function of severity of CHF. PMID- 17427385 TI - Transthoracic echocardiographic measurement of patent ductus arteriosus in dogs. AB - BACKGROUND: Patent ductus arteriosus (PDA) size and morphology influence the selection of the kind and the size of the embolization device used to effect shunt closure. HYPOTHESIS: That echocardiographic measurement of PDA in dogs is accurate. ANIMALS: Forty-five client-owned dogs with PDA. METHODS: Prospective observational study. Echocardiographic and angiographic data were compared. RESULTS: Measurement of the ductus in color Doppler echocardiography (CD-E) and 2 dimensional echocardiography (2D-E) was achieved from left parasternal views in 43 of 45 unsedated dogs (96%). In these 43 dogs, the angiographic minimal PDA diameter was 3.72 +/- 1.59 mm, and the diameter of the PDA ampulla was 8.46 +/- 3.01 mm. The CD-E minimal PDA diameter ranged from 2.3 to 9.5 mm (median, 4.0 mm). There was a significant mean difference to the angiographic measurements (1.15 +/- 0.95 mm; P < .0001). An agreement in a 1-mm range was found in 21 of 43 dogs (48%). The 2D-E minimal PDA diameter was 3.73 +/- 1.78 mm, and the mean difference to the angiographic measurements was not significant (0.00 +/- 0.72 mm; P = .98). An agreement in a 1-mm range was found in 31 of 43 dogs (72%). The 2D-E measurement of the PDA ampulla revealed a significant mean difference to the angiographic data (1.95 +/- 2.43 mm, P < .0001). An agreement in a 2-mm range was found in 21 of 43 dogs (49%). CONCLUSIONS AND CLINICAL IMPORTANCE: The 2D-E from the left cranial parasternal view is an excellent noninvasive method to estimate the PDA minimal diameter before doing catheter intervention. PMID- 17427386 TI - Chordae tendineae rupture in dogs with degenerative mitral valve disease: prevalence, survival, and prognostic factors (114 cases, 2001-2006). AB - BACKGROUND: Degenerative mitral valve disease (MVD) is the most common heart disease in small breed dogs, and chordae tendineae rupture (CTR) is a potential complication of this disease. The survival time and prognostic factors predictive of survival in dogs with CTR remain unknown. HYPOTHESIS: The prevalence and prognosis of CTR in dogs with MVD increases and decreases, respectively, with heart failure class. ANIMALS: This study used 706 dogs with MVD. METHODS: The diagnosis of CTR was based on a flail mitral leaflet with the tip pointing into the left atrium during systole, which was confirmed in several 2-dimension imaging planes using the left and right parasternal 4-chamber views. RESULTS: CTR was diagnosed in 114 of the 706 dogs with MVD (16.1%) and most of these (106/114, 93%) had severe mitral valve regurgitation as assessed by color Doppler mode. CTR prevalence increased with International Small Animal Cardiac Health Council (ISACHC) clinical class (i.e., 1.9, 20.8, 35.5, and 69.6% for ISACHC classes Ia, Ib, II, and III, respectively [P < .05]). Long-term follow-up was available for 57 treated dogs (angiotensin-converting enzyme inhibitors and diuretics) and 58% of these (33/57) survived > 1 year after initial CTR diagnosis (median survival time, 425 days). Clinical class, the presence of ascites or acute dyspnea at the time of diagnosis, heart rate, plasma urea concentration, and left atrial size were predictors of survival. CONCLUSIONS AND CLINICAL RELEVANCE: CTR is associated with a higher overall survival time than previously supposed. Its prognosis mostly depends on a combination of clinical and biochemical factors. PMID- 17427387 TI - Omega-3 fatty acids in Boxer dogs with arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Omega-3 fatty acids have been shown to reduce arrhythmia in animal models and people. These effects have not been studied in dogs with spontaneously occurring arrhythmia. HYPOTHESIS: Fish oil will reduce the frequency of ventricular arrhythmia in Boxer dogs with arrhythmogenic right ventricular cardiomyopathy (ARVC). ANIMALS: Twenty-four Boxers with ARVC were included in this study. METHODS: Asymptomatic Boxers not receiving antiarrhythmic medications were evaluated with echocardiogram and electrocardiogram. Dogs with at least 1 ventricular premature contraction (VPC) received 24-hour ambulatory electrocardiography (AECG) recordings. Dogs with > 95 VPCs in 24 hours were randomized to 1 of 3 treatments: (1) Fish oil, 2 g; (2) Flax oil, 2 g; or (3) sunflower oil, 2 g (Control group), for 6 weeks. Investigators and owners were blinded to the treatment groups. All baseline measurements were repeated after the 6-week supplementation. RESULTS: There were no differences at baseline for age, sex, blood pressure, weight, echocardiographic measurements, or VPCs. Median number of VPCs in 24 hours for all dogs was 543 (range, 96-40,063) at baseline and 193 (range, 6-14,825) after 6 weeks of supplementation. VPCs/24 h were reduced for the Fish oil group (baseline median = 397 [range, 249-10,587]; 6-week median = 162 [range, 16-3,781]; P = .02), but not for the Flax oil (P = .58) or Control (P = .48) groups. CONCLUSIONS AND CLINICAL IMPORTANCE: These data suggest that fish oil, but not flax oil, supplementation for 6 weeks reduces arrhythmia in Boxers with ARVC and that it could be useful in treating this common disease. Further studies are needed to determine optimal dose and duration of treatment. PMID- 17427388 TI - Clinical stage, therapy, and prognosis in canine anal sac gland carcinoma. AB - BACKGROUND: Reports of canine anal sac gland carcinoma (ASGC) describe varied clinical presentations and management and differing responses to therapy. A unifying approach to clinical stage determination and management of this disease has yet to be presented. HYPOTHESIS: An ordinal clinical staging scheme for canine ASGC can be devised on the basis of responses to therapy for a retrospective cohort of affected dogs. ANIMALS: 130 dogs with naturally occurring ASGC. METHODS: A simplified clinical stage system and a management algorithm for canine ASGC were derived from retrospective evaluation of a cohort of 80 dogs; applicability of both was then prospectively evaluated in a cohort of 50 dogs. RESULTS: Retrospective evaluation revealed 4 statistically significant negative prognostic indicators for survival: lack of therapy, presence of distant metastases, presence of lymph node metastases, and primary tumor size. Lymph node extirpation was a statistically significant positive prognostic indicator by bivariate analysis. In both retrospective and prospective analyses, the modified clinical stage scheme revealed a significant association with survival time. CONCLUSIONS AND CLINICAL IMPORTANCE: The clinical staging scheme permits differentiation between groups in terms of prognosis and, therefore, decisions on therapy. This will facilitate application of appropriate therapy and enhanced communication and collaboration in further investigations of ASGC. PMID- 17427389 TI - Canine Dal blood type: A red cell antigen lacking in some Dalmatians. AB - BACKGROUND: Based upon alloantibodies produced after sensitizing dogs with transfused blood, more than a dozen blood group systems have been recognized thus far, and some have been classified as dog erythrocyte antigens (DEA). HYPOTHESIS: A new canine red cell antigen was suspected, based on the development of specific alloantibodies in a Dalmatian previously sensitized by blood transfusions. ANIMALS: Twenty-six Dalmatians (including 1 Dalmatian in need of blood compatibility studies); 55 canine blood donors. METHODS: Serologic tests, including blood typing, crossmatching, and direct Coombs' test were performed by standard tube techniques and a novel gel column technology adapted from human blood banking. RESULTS: By day 40 after transfusion of an anemic Dalmatian, all major crossmatch tests to 55 non-Dalmatian dogs were incompatible. The 2 initial donors, who were compatible before transfusion, were also now incompatible, suggesting the development of an alloantibody to a common red cell antigen. No siblings were available, but 4 of 25 unrelated Dalmatians were crossmatch compatible, suggesting that they were missing the same red cell antigen. The patient was blood typed DEA 1.1, 3, 4, and 5 positive, but DEA 7 negative. Further blood typing and crossmatching results did not support an association to any of these known blood types. The alloantibodies produced were determined to be of the immunoglobulin G class. CONCLUSIONS AND CLINICAL IMPORTANCE: Based upon the identification of an acquired alloantibody in a Dalmatian, a presumably new common blood type named Dal was identified. Dalmatians lacking the Dal antigen are likely at risk of delayed and acute hemolytic transfusion reactions. PMID- 17427390 TI - A newly recognized blood group in domestic shorthair cats: the Mik red cell antigen. AB - BACKGROUND: Naturally occurring alloantibodies produced against A and B red cell antigens in cats can cause acute hemolytic transfusion reactions. Blood incompatibilities, unrelated to the AB blood group system, have also been suspected after blood transfusions through routine crossmatch testing or as a result of hemolytic transfusion reactions. HYPOTHESIS: Incompatible crossmatch results among AB compatible cats signify the presence of a naturally occurring alloantibody against a newly identified blood antigen in a group of previously never transfused blood donor cats. The associated alloantibody is clinically important based upon a hemolytic transfusion reaction after inadvertent transfusion of red cells expressing this red cell antigen in a feline renal transplant recipient that lacks this red cell antigen. METHODS: Blood donor and nonblood donor cats were evaluated for the presence of auto- and alloantibodies using direct antiglobulin and crossmatch tests, respectively, and were blood typed for AB blood group status. Both standard tube and novel gel column techniques were used. RESULTS: Plasma from 3 of 65 cats and 1 feline renal transplant recipient caused incompatible crossmatch test results with AB compatible erythrocytes indicating these cats formed an alloantibody against a red cell antigen they lack, termed Mik. The 3 donors and the renal transplant recipient were crossmatch-compatible with one another. Tube and gel column crossmatch test results were similar. CONCLUSIONS AND CLINICAL IMPORTANCE: The absence of this novel Mik red cell antigen can be associated with naturally occurring anti-Mik alloantibodies and can elicit an acute hemolytic transfusion reaction after an AB-matched blood transfusion. PMID- 17427391 TI - Determination of lactate concentrations in blood plasma and peritoneal fluid in horses with colic by an Accusport analyzer. AB - BACKGROUND: Intestinal hypoperfusion can lead to increased lactate concentrations in plasma and peritoneal fluid of horses with colic. HYPOTHESIS: The purposes of this study were to (1) evaluate the reliability of the Accusport analyzer to assess peritoneal fluid lactate (PFL) concentrations in healthy horses and those with colic, (2) identify clinical features associated with abnormal blood plasma lactate (BPL) and PFL concentrations, and (3) evaluate the prognostic value of BPL and PFL. ANIMALS: BPL and PFL were determined in 20 healthy horses and in 106 horses with colic. RESULTS: The Accusport was reliable for determining BPL concentrations < 13 mM and PFL concentrations < 20 mM. Multivariate analysis indicated that PCV and the need for intestinal resection were independently associated with the BPL; pulse, PCV, venous pO2, the presence of necrotic intestine, an increased amount of peritoneal fluid, and fluid total protein content were independently associated with PFL. With a 1 mM increase in BPL or PFL, the respective odds ratios for required abdominal surgery increase to 1.23 (BPL) and 1.58 (PFL), odds ratios for a required intestinal resection increase to 1.20 (BPL) and 1.41 (PFL), and odds ratios for developing ileus increase by 1.33 (BPL) and 1.36 (PFL). PFL concentrations of 1, 6, 12, and 16 mM correspond to a probability of death of 11, 29, 63, and 82%, respectively, in horses without strangulating obstruction and of 25, 52, 82, and 92%, respectively, in horses with strangulating obstruction. CONCLUSION: PFL is more useful and sensitive than BPL for prognostic purposes in horses with colic. PMID- 17427392 TI - Cardiovascular effects of acute pulmonary obstruction in horses with recurrent airway obstruction. AB - BACKGROUND: Recurrent airway obstruction (RAO) is common in horses. Although pulmonary artery (PA) pressure increases during RAO, cardiac function in horses with RAO has received limited attention. HYPOTHESIS: The purpose of this study was to noninvasively determine the cardiovascular effects of acute pulmonary obstruction (APO) in horses with RAO and their reversibility. ANIMALS: Five geldings with RAO, inducible by exposure to moldy hay, were studied. METHODS: Pulmonary mechanics, echocardiography, serum troponin I concentrations, arterial blood gases, and hematocrit were obtained before and after 7 days of APO. Heart rate, PA diameter and flow characteristics, right and left ventricular luminal dimensions and wall thicknesses, global cardiac performance, and evidence of myocardial damage were evaluated. Pulmonary mechanics and echocardiography were reevaluated during remission. RESULTS: [corrected] Severe, transient APO did not induce chronic cor pulmonale in horses, because cardiac anatomy and function were normal between episodes. An acute episode of APO produced anatomical and functional cardiac changes in both the right and left heart (including increased PA diameter, abnormal septal motion, and decreased left ventricular diameter and estimated stroke volume), possibly because of the development of pulmonary hypertension, without apparent myocardial damage. The decrease in stroke volume was offset by the increase in heart rate. CONCLUSIONS AND CLINICAL IMPORTANCE: With APO of 7 days' duration, cardiovascular abnormalities and the functional airway changes that produce them are reversible when the offending allergens are removed. PMID- 17427393 TI - Detection of fibrin deposits in tissues from horses with severe gastrointestinal disorders. AB - BACKGROUND: In humans and experimental animals, disseminated intravascular coagulation (DIC) causes fibrin deposition in several organs, which eventually leads to ischemia and multiorgan failure. HYPOTHESIS: Horses who died or were euthanized for severe gastrointestinal disorders could have fibrin deposits in different tissues. ANIMALS: Tissue-organ samples collected during postmortem examinations on 66 colic horses with poor prognoses (eg, severe intestinal ischemia, enteritis, peritonitis), from 11 colic horses with good prognoses (eg, large-colon obstruction or displacement), and from 16 slaughter horses. METHODS: Tissue samples (kidney, lung, liver) were stained with hematoxylin and eosin, and phosphotungstic acid hematoxylin for a blinded histologic examination. A fibrin score (grades 0 to 4) was established for each tissue sample and for each horse. RESULTS: Fibrin deposits were found in tissue specimens of 11 of 27 of horses (40.7%) in the ischemic group, 8 of 21 in the enteritis group (38.1%), and 7 of 18 in the peritonitis group (39.0%), whereas none of the horses in the obstructive group (n = 11) and only 1 horse in the slaughter group (n = 16) had fibrin deposits in their tissues. In addition, the mean fibrin score values for the ischemic, enteritis, and peritonitis groups (1.3 +/- 1.7, 1.1 +/- 1.6, and 0.9 +/- 1.3, respectively) were statistically higher than those for the obstructive and slaughter groups (0.0 +/- 0.0 and 0.1 +/- 0.5, respectively). The largest fibrin deposits were found in the lungs. CONCLUSIONS AND CLINICAL IMPORTANCE: Horses with severe gastrointestinal disorders have fibrin deposits that are consistent with capillary microthrombosis, multiorgan failure, and DIC. PMID- 17427394 TI - Synthetic adrenocorticotropic hormone stimulation tests in healthy neonatal foals. AB - BACKGROUND: Cosyntropin (adrenocorticotropic hormone [ACTH]) stimulation tests are used to evaluate adrenal function. Low-dose ACTH stimulation tests are the most accurate method for diagnosing relative adrenal insufficiency in critically ill humans but have not been evaluated in foals. HYPOTHESIS: Peak serum cortisol concentrations in healthy foals will not be significantly different after intravenous administration of 1, 10, 100, and 250 microg of cosyntropin. ANIMALS: 14 healthy neonatal foals, 3-4 days of age. METHODS: A randomized cross-over model was used in which cosyntropin (1, 10, 100, or 250 microg) was administered intravenously on days 3 and 4 of life. Blood samples were collected before and 30, 60, 90, 120, and 150 minutes after administration of cosyntropin for determination of serum cortisol concentration. RESULTS: Serum cortisol concentrations did not significantly increase after administration of 1 microg of cosyntropin. Cortisol concentration peaked 30 minutes after administration of 10 microg of cosyntropin and 90 minutes after 100 and 250 microg of cosyntropin. There was no relationship between cosyntropin dose and serum cortisol concentration at 30 minutes. Compared with the 10-microg dose, 100 and 250 microg of cosyntropin induced significantly greater cortisol concentrations at 90 minutes, at which point the 10-microg cosyntropin-dose cortisol values were indistinguishable from baseline. There was no significant difference in the area under the cortisol concentration curve between the 100- and 250-microg doses. No effect of day of testing or foal weight on peak cortisol concentration was detected. CONCLUSIONS AND CLINICAL IMPORTANCE: The results of this study suggest that 10- and 100-microg doses of cosyntropin would be appropriate for evaluating adrenal function in neonatal foals. PMID- 17427395 TI - Compressive cervical myelopathy in young Texel and Beltex sheep. AB - BACKGROUND: This investigation was prompted by the referral of increasing numbers of young Texel and Beltex rams with ataxia and weakness, or wobbler syndrome. HYPOTHESIS: The study aims were to describe the clinical and pathologic findings in affected sheep. ANIMALS: The animals evaluated in this study included 7 Texel sheep (6 male and 1 female) and 3 Beltex sheep (2 male and 1 female) referred from pedigree flocks. Typically, the sheep were 15-18 months of age at referral. METHODS: Diagnostic investigations included radiographic and computed tomographic (CT) myelography followed by gross postmortem and histopathologic examinations. RESULTS: Clinical findings typical of cervical spinal cord compression were present in all sheep but varied in severity. Myelography confirmed dorsal spinal cord compression in the region of C6-C7. No bony abnormalities were identified as described in cases of canine and equine wobbler syndrome. Postmortem examinations revealed discrete, smooth, nodular to polypoid projections of adipose tissue apparently prolapsing through the dorsolateral intervertebral space at C6-C7 and causing localized spinal cord compression. Histopathology of the nodules confirmed that they were composed of well-differentiated adipocytes typical of fatty tissue. Spinal cord lesions were similar in all sheep with marked Wallerian degeneration at the site of compression and mild Wallerian degeneration present cranial and caudal to the lesion. CONCLUSIONS AND CLINICAL IMPORTANCE: The findings of this study suggest a novel cervical myelopathy in these sheep breeds caused by the presence of fatty nodules encroaching into the dorsal vertebral canal at C6-C7. Additional investigations are required to establish the etiology and possible hereditary risk factors for this unique clinicopathologic syndrome. PMID- 17427396 TI - Neurotoxicosis in 4 cats receiving ronidazole. PMID- 17427397 TI - Hyperthyroidism associated with probable struma cordis in a young dog. PMID- 17427398 TI - Presumptive fluphenazine-induced hepatitis and urticaria in a horse. PMID- 17427399 TI - Multicentric mastocytoma in a horse. PMID- 17427400 TI - Disseminated intravascular coagulation in a horse with Streptococcus equi subspecies zooepidemicus meningoencephalitis and interstitial pneumonia. PMID- 17427401 TI - Ultrasonographic diagnosis of aortoiliac thrombosis in 2 calves. PMID- 17427402 TI - Genotypic analysis of Giardia duodenalis in domestic cats. AB - BACKGROUND: Giardia duodenalis is an intestinal flagellated protozoan that affects many mammalian species often causing severe diarrheal disease. Several different genotypes have been identified (Assemblages A-G). Most isolates recovered from domestic cats have been assigned to either Assemblage A, the zoonotic form of the parasite, or Assemblage F, identified thus far only in cats. Genotypic variation within G. duodenalis may influence clinical presentation and course of disease. Therefore, host-adapted genotypes may not be responsible for diarrheal disease (eg, Assemblage F in cats). HYPOTHESIS: Multiple Giardia genotypes will be present in domestic cats, including Assemblage F, which will not be correlated with clinical signs. ANIMALS: 250 domestic cats from eastern Mississippi and northwestern Alabama. METHODS: Prevalence survey. Fecal samples evaluated for cysts using a centrifugation concentration technique and a commercially available direct immunoflourescent antibody kit. Giardia isolates were characterized by PCR amplification and sequencing of the glutamate dehydrogenase gene. RESULTS: Both Assemblage A-I (6/17) and Assemblage F (11/17) were identified. Although Assemblage was significantly associated with age and housing, no association was detected between Assemblage and a variety of other factors including the presence of gastrointestinal signs (acute vomiting, diarrhea, and constipation). CONCLUSIONS AND CLINICAL IMPORTANCE: The presence of diarrhea in domestic cats with Giardia cannot be used as a predictor of the presence of zoonotic genotypes in animals within the study area. Although Assemblage A was associated with age and housing, veterinarians should consider any isolation of Giardia from domestic cats as potentially zoonotic. PMID- 17427403 TI - Inflammatory airway disease of horses. AB - The purpose of this consensus statement is to provide a review of current knowledge and opinions concerning inflammatory airway disease (IAD) and to help practitioners differentiate IAD from heaves (or recurrent airway obstruction; RAO) and other inflammatory respiratory diseases of horses. PMID- 17427404 TI - [Elaboration of systematic position and some physiological characteristics of collection strains of Debaryomyces hansenii (Zopf) Looder et Kreger-Van Rij]. AB - Morphological and physiological properties of 52 strains of yeast Debaryomyces hansenii from the Ukrainian collection of microorganisms has been investigated. The belonging of 40 strains to the given species is confirmed. According to new classification 35 of them are referred to D. hansenii var. hansenii, five strains -to D.hansenii var. fabryi. It is shown, that assimilation of hydrocarbons and antagonistic properties are inherent in a small amount of strains. One half of the investigated debaryomycetes are riboflavin producers. The osmo- and galofility is inherent in all the investigated strains. These characteristics are proposed to be applied as additional ones in identification of these yeast. PMID- 17427405 TI - [Purple sulfur bacteria isolated from reservoirs of the Yavoriv sulfur deposit]. AB - Three pure cultures of purple sulfur bacteria were isolated from reservoirs of the Yavoriv sulfur deposit. The studying of their morphology, cytology and physiology has confirmed the belonging of these bacteria to Chromatiaceae family and has allowed identifying them as Thiocapsa sp., Lamprocystis sp. and Chromatium sp. PMID- 17427406 TI - [Ethanol into acetaldehyde bioconversion by mutant strains of Hansenula polymorpha Felcao de Morais & Dalia Maia]. AB - The influence of some factors (tris-HCl buffer, pH 8.0, concentration, use of different binding agents aeration modes, genetically determined peroxisome degradation damage) on biotransformation efficiency of 0.217 M (1%) ethanol to acetaldehyde at 30 degrees C by Hansenula polymorpha 7-4A (gcrl EAO) strain cells with glucose repression block was investigated. Optimal cultivation conditions for cells were selected. Bioconversion efficiency using 1 M tris-HCl buffer, pH 8.0, was found the highest one as compared with using the buffer in concentrations from 0.1 M to 3 M. The process efficiency when using tris(hydroxymethyl)aminomethane as binding acetaldehyde agent proved much higher than when using sodium bisulfite both at aeration by air stream and incubation on shaker. Using 146 and 179 mutants cells for bioconversion with defects in alcohol oxidase inactivation during macropexophagy stimulated efficiency increase by 5.58% and 8.10%, respectively, as compared with the use of parental 7-4A strain cells. PMID- 17427407 TI - [Characteristics of carotinoids of methylotrophic bacteria of Methylobacterium genus]. AB - A detailed chromatographic and spectrophotometric analysis of carotinoids in Methylobascterium genus strains: M. extorquens B-3362, M. fujisawaense B-3365, M. mesophilicum B-3352. It has been shown that carotinoids of the studied strains are mainly represented by xanthophylls. The carotinoid common for all three strains and presented in the highest quantity was identified as oscilloxanthin. Carotene found in M. fujisawaense B-3365 and M. mesophilicum B-3352 species was identified as 3,4-didehydrolycopene. Quantitative content of carotenoids was 0.45 mg/g of dry weight for M. fujisawaense B-3365 and M. mesophilicum B-3352 and 0.23 mg/g for M. extorquens B-3362. Interspecies differences were revealed both in qualitative and quantitative content of carotinoids. However, in spite of such differences, carotinoids are similar in these three strains by Rf in thin-layer chromatography and by output time in highly efficient liquid chromatography carotinoids, and make about 80% of the total content of carotinoids. PMID- 17427408 TI - [Similar nucleotide blocks in tobamoviral subgenomic promoters]. AB - It was discovered by computational analysis of nucleotide sequences that the movement (MP) and coat proteins (CP) subgenomic RNA promoter regions of the most tobamoviruses tested (10 from 15) contain almost identical three-component nucleotide blocks. In the middle part of the block there is the central nine nucleotide motive GGTTCGTTT or GATTCGTTT, flanged with length-variable GC- and AT containing sequences. GC-containing sites are localized upstream of the central motive and possess the CGC motive. The sixth nucleotide of central motives (G) is located in genomes 65-81 nucleotides upstream of MP gene or 9-20 nucleotides upstream of CP gene. The others tobamoviruses tested (5 from 15) have a considerable quantity of nucleotide replacements in central motives, high variability of GC-containing sites as well as two similar nucleotide blocks in promoters of CP genes. The role of the similar nucleotide blocks as functional elements of tobamoviral subgenomic RNA promoters is discussed. PMID- 17427409 TI - [Amplification of proviral DNA of bovine immunodeficiency virus in experimentally infected animals]. AB - The primer set for detection of bovine immunodeficiency virus (BIV) proviral DNA, retrovirus of unknown pathology, by standard polymerase chain reaction was developed. Amplicon of the expected size was detected in DNA from peripheral blood mononuclear cells of seropositive experimentally BIV infected sheep and cattle. Primers targeted the short fragment of BIV pol gene. Sequences of BIV proviral DNA extracted from BIV infected cell culture as well as from experimentally infected cows were taken for targeted pol BI BPX gene locus. Problems of BIV detection from the clinical material of the experimentally and naturally infected animals are discussed. PMID- 17427410 TI - [Pantoea agglomerans--pathogen of Elytrigia repens and Arrhenatherum elatius diseases]. AB - Strains of Pantoea agglomerans were isolated from the samples with symptoms of bacterial damage of various organs of Elytrigia repens and Arrhenatherum elatius. The isolated bacteria are aggressive to different extent for a host-plant and a number of cultivated plants. Some problems of evolution of this bacterial species parasitism are discussed. PMID- 17427411 TI - [Phosphinothricin-resistant somatic hybrids Brassica napus + Orychophragmus violaceus]. AB - Phosphinothricin (PPT) resistant hybrid plants between Brassica napus L. cv. Kalinovsky and Orychophragmus violaceus (L.) O.E. Shulz. were obtained as a result of somatic hybridization experiments. The hybrids inherited PPT resistance from O. violaceus plants which were previously transformed by the vector containing Spm/dSpm Zea mays transposon system with bar gene located within the nonautonomous transposon. The obtained plants had intermediate morphology. Their hybrid nature has been confirmed by isozyme (esterase and amilase activity) and PCR (bar, gus, Spm/dSpm integration) analyses. The hybrids combined B. napus plastom and O. violaceus mithochondrion that was revealed by PCR-RFLP. The hybrid plants might be included to rapeseed breeding programme after examination of their oil quality as well as to chloroplast transformation experiments that is still urgent for B. napus. PMID- 17427412 TI - [The role of Cd-binding proteins and phytochelatins in the formation of cadmium resistance in Nicotiana plumbaginifolia cell lines]. AB - Nicotiana plumbaginifolia callus lines with the equal resistance to cadmium have been produced under different selective conditions--either without inhibition of the phytochelatin synthesis (line Cd-R) or in the presence of the inhibitor butionine sulfoximine (line Cd-Ri). The level of phytochelatin synthesis in the line Cd-R five-fold exceeded the control value and in the line Cd-Ri it was twice as much as in the control. It was shown that in the control line mainly three cadmium-binding proteins are expressed of the molecular weihgts 41, 34 and 19 kD. The common feature of the both resistant lines is the expression of the cadmium binding proteins of 40, 37 and 19 kD. The resistant lines differ with respect to the synthesis of relatively low-molecular cadmium-binding proteins. The proteins of the molecular weights 12.5, 11.5 and 9 kD are expressed in the line Cd-R, while the proteins of 13 and 10 kD are expressed in the line Cd-Ri. It was supposed that both the phytochelatins and the Cd-binding proteins contribute to the resisitance of N. plumbaginifolia callus lines to cadmium and the lack of the phytochelatins can be equilibrated by the changes in the low-molecular Cd-binding protein synthesis. PMID- 17427413 TI - [Mobilization of simazine degradation genes by the plasmid pSa]. AB - The plasmid pSa was found to mobilize the genes for simazine degradation (smz) of the rhizosphere bacterium Herbaspirillum sp.B601 by forming hybrid pSa-Smz plasmids. Independent migration of smz genes into various loci of genome during transfer and elimination of the hybrid plasmids indicated that the genes were parts of a "catabolic island" which could be unstable under certain conditions. PMID- 17427414 TI - Telomeres and seed banks. AB - We have found that a progressive loss of telomeric sequences occurs from high molecular weight DNA with an increasing appearance at low molecular weight as the periods of storage in the dry state were extended in time to provide seed germination loss from 98 to 0%. Telomere distribution would appear to follow the general pattern of DNA random fragmentation that occurs in the embryos of seeds stored in the dry state, but there are also indications of an overall telomere loss from DNA as a consequence of storage. There is a need for a convenient "equality marker" for the seeds that can be monitored over time. Reviewing the implications of our results very carefully we believe that there is considerable potential in the use of telomere sequences to mark embryo ageing of seeds held in Seed Banks. PMID- 17427415 TI - [Structural and functional organization of mitochondria in soybean root statocytes under microgravitation]. AB - The effects of microgravity and ethylene on morphology and ultrastructural organization of mitochondria in root statocytes of soybean seedlings grown for 6 days on the board of the space shuttle Columbia during the STS-87 mission were investigated. The spaceflight seedlings and the ground-grown control seedlings were grown in BRIG (Biological Research in Canister) in the presence of KMnO4 to remove ethylene. It was revealed that irrespectively of KMnO4 treatment the mitochondria in the spaceflight seedlings were characterized by round or oviform and by low electron density of organelle matrix, whereas the organelles in the ground controls were polymorphic in shape and had higher electron density of matrix. The possible mechanisms of morphological and ultrastructural rearrangements of mitochondria that may be involved in adaptation processes of soybean seedlings to microgravity conditions are discussed. PMID- 17427416 TI - Comparison of microarray and sage techniques in gene expression analysis of human glioblastoma. AB - To enhance glioblastoma (GB) marker discovery we compared gene expression in GB with human normal brain (NB) by accessing SAGE Genie web site and compared obtained results with published data. Nine GB and five NB SAGE-libraries were analyzed using the Digital Gene Expression Displayer (DGED), the results of DGED were tested by Northern blot analysis and RT-PCR of arbitrary selected genes. Review of available data from the articles on gene expression profiling by microarray-based hybridization showed as few as 35 overlapped genes with increased expression in GB. Some of them were identified in four articles, but most genes in three or even in two investigations. There was found also some differences between SAGE results of GB analysis. Digital Gene Expression Displayer approach revealed 676 genes differentially expressed in GB vs. NB with cut-off ratio: twofold change and P < or = 0.05. Differential expression of selectedgenes obtained by DGED was confirmed by Northern analysis and RT-PCR. Altogether, only 105 of 955 genes presented in published investigations were among the genes obtained by DGED. Comparison of the results obtained by microarrays and SAGE is very complicated because authors present only the most prominent differentially expressed genes. However, even available data give quite poor overlapping of genes revealed by microarrays. Some differences between results obtained by SAGE in different investigations can be explained by high dependence on the statistical methods used. As for now, the best solution to search for molecular tumor markers is to compare all available results and to select only those genes, which significant expression in tumor combined with very low expression in normal tissues was reproduced in several articles. 105 differentially expressed genes, common to both methods, can be included in the list of candidates for the molecular typing of GBs. Some genes, encoded cell surface or extra-cellular proteins may be useful for targeting gliomas with antibody-based therapy. PMID- 17427417 TI - [Genetic structure of diploid-polyploid complex of spined loaches Cobitis (Cypriniformes, Cobitidae) of the Danube delta]. AB - The unusually high diversity of spined loache biotypes in Lower Danube was detected by means of biochemical genetic investigation and cytometric analysis of 358 specimens collected in riverbed and eriks. Along with two diploid species (C. elongatoides and C. "tanaitica") six hybrid forms were revealed: diploid C. elongatoides "tanaitica"; triploid C. 2 elongatoides--"tanaitica", C. elongatoides--2 "tanaitica" and C. 2 elongatoides--species-1 and tetraploid C. 3 elongatoides--"tanaitica", C. elongatoides--species-2--2 "tanaitica". Besides that the specimens with recombinant genotypes occured. In spite of the apomictic mode of reproduction the poliploids do not possess the clonal structure but according to the level of polymorphism and the genotype distribution are isomorphous to the parental diploid species. This means that on the contrary to the polyploid cobitids of the Dnieper which have appeared in this catchment area due to the expansion of their home range the polyploid fishes from the Lower Danube reaches are autochtonous and are produced as a result of hybridization with the local diploid species. The process is seemingly going on without any kind of limitations. PMID- 17427418 TI - [Using chromosome aberration frequency analysis in formation of risk groups for neoplastic diseases]. AB - Taking into account that instability of chromosomes represents the mutagenic influence of of environmental factors; that chromosomal aberrations can be used as the markers of gene changes (including the oncologic ones); that the absence of the well-defined answer for age dependence on frequency of aberrations of the inspected persons; for oncologic patients sum up the promoted level of this index: on the basis of determination of index "frequency of aberrations of chromosomes" it is possible to form groups of the promoted risk, including oncologic for more deep study other methods. PMID- 17427419 TI - [Genetic and biochemical aspects of keratin synthesis by hair follicles]. AB - In this review article the data about synthesis and gene regulation of keratin by hair follicles have been summarized. It has been shown that both differentiation of hair follicle matrix cells and normal growth of hair require the coordinated activities of the genes encoding structural proteins. The keratin genes are clustered in families and are usually 5-10 kb in the genome. The separate clusters of two keratin IF gene families and five KAP gene families have been discovered and some of them have been mapped. The close relation between these clusters suggests that the "global" regulatory domains might govern their expression. PMID- 17427420 TI - [State management of health care in Ukraine: theory and practice]. AB - The authors consider in the article the theory and practice of state management of health care of Ukraine. PMID- 17427421 TI - [Pilote strategy to optimize an individual treatment of arterial hypertention and algorithm of the prophylactics of vascular complications]. AB - Ischemic stroke (IS) and myocardial infarction are very serious complications of arterial hypertention (AH). To prevent such complications it is important to control the level of arterial pressure, state of lipid spectre and to reveal hypercoagulation in blood which is a predictor to the formation of thrombi and after its severity degree to find clinical homeostatic risk factor, individual dose of an antiaggregant medication and its application. 146 patients with II-III stage arterial hypertention were observed using "Koaguloscop-TC" apparatus. 36 parameters reflecting the formation of all phases of coagulation cascade and the fibrinolysis system of these patients have been studied. To optimize individual treatment of arterial hypertention and prophylactics of vascular complications is necessary to prescribe a complex treatment including antihypertensive, hypolypidemic and antiagragative agents. PMID- 17427422 TI - [Structural and functional stage of the myocardium in different forms of ischemic heart disease]. AB - 152 patients with ischemic heart disease aged from 46 to 60 years have been observed. Among these patients, 69 were with stable stenocardia, 32--with unstable stenocardia and 53--in stage related to acute heart attack. Normal geometry of the heart was found in 33% of patients with stable stenocardia, excentric hypertrophy was found in 30% of patients with stable stenocardia. Excentric hypertrophy prevailed in patients with unstable stenocardia and myocardial infarction. Systolic disfunction of the myocardium was found in 72% of patients with stable stenocardia, in 41% of patients with unstable stenocardia and in 43% of patients with myocardial infarction. Diastolic dysfunction of the myocardium in terms of rigide and restrictive types was found oftener in patients with myocardial infarction. PMID- 17427423 TI - [Functional state of the vessel endothelium in patients with systemic lupus erythematosis]. AB - SLE patients show the formation of endothelial dysfunction manifested by the decreuse of endothelial dependent vasodilatation in response to reactive hyperemia test and incapuse of carotid artery intima-media thickness (IMT). Disorder of Endothelial dependent vasodilation damage in SLE patients is associated with age, damage index and inflammatory process activity. An association with the duration of the disease was not been found. (IMT) thickness increase is associated with the age, damage index and has no correlation with the duration of the disease. The rate of atherosclerotic plaques in the carotid artery in SLE patients exceeds by 1.8 times that of the control group. Atherosclerotic identification rate was found to have tight association with age, disease activity and damage index. Atherosclerotic damage degree is associated with damage index and disease activity and has no correlation with age and the duration of the disease. PMID- 17427424 TI - [Index state of contact platelet activation in patients with acute viral myocarditis]. AB - The article shows that ISCART statistically correlates with the degree of severity of acute virus myocarditis. The authors proposed for the first time the group of medications and proved that efficiency of these medications correlates with the degree of severity of the disease and has a place to be used by physicians. PMID- 17427425 TI - [Metabolic syndrome in men of 35-65 years old from rural district of Poltava region and possibility of the prophylactic therapy with atorvastatine]. AB - Observation of 250 men aged 35-65 years (middle age was 52 +/- 2 years) showed that prevalence of MS was 27%. Atorvastatine administration at dose of 10-20 mg per day for patients with ischemic heart disease and chronic obstructive lung disease associated with MS reduced during month administration the level of cholesterol, triglycerides, beta- and pre-beta-lipoproteids and increased the level of alpha-cholesterol in blood of these patients. Addition of atorvastatine to a common treatment patients with chronic obstructive lung disease improved as well the clinical picture of systemic inflammation process. PMID- 17427426 TI - [Treatment of patients with duodenal peptic ulcer not associated with Helicobacter infection]. AB - The authors have shown in the artcile personality characteristics and neuropsychic disorders in patients with duodenal peptic ulcer. These characteristics are the following: increased level of anxiety, presence of frustration, agressiveness, apparent rigidity and astenic, depressive, affective derangements. It does have not positive influence on the course of the disease and requires adequate correction. Psychotherapy use in a complex treatment makes for ulcers healing, dicreasing treatment period and numbers of relapses. It eliminates disorders of psychoemotional status of patients, enhances quality of life of patients from the beginning of the treatment. PMID- 17427427 TI - [Functional state of the autonomic nervous system in young persons with gastroesophageal reflux disease]. AB - The article considers changes of the vegetative nervous system in young persons with gastroesophageal reflux disease (GERD). The author has analyzed prevalence of vegetative dysfunctions in young persons with GERD. Clinical pictures and grade of severity of the disease have been outlined. Individual instrumental assessment of a vegetative tone and vegetative reactivity has been carried out. Obtained data show that functional disorders of the vegetative nervous system are leading factors in the genesis of GERD in young persons. PMID- 17427428 TI - [Prognostic and diagnostic criteria in the development of bronchial asthma in children of young school age]. AB - The analyzes of anthropometric parameters, somatotype and dermatoglyphics of children aged 7-12 years with bronchial asthma have been studied. Obtained results have shown that somatotype and dermatoglyphics are specific genetic and phenotype markers of susceptibility to the development of bronchial asthma. PMID- 17427429 TI - [Soy-based food in a complex treatment of patients with tuberculosis]. AB - Results of clinical and laboratoty studies on efficiency and safety of upplied soya-based food during intensive antimicobacterial treatment of patients with tuberculosis are presented in the article. Obtained results showed the necessity of introduction soya-based food in a complex treatment of this infection as it enhances processes of detoxication, positevely influences on functional and structural state of the liver, reduces cytolysis, cholestasis symptomes, mesenchymal and inflamation reactions thus increases the tolerance of antimicobacterial agents. PMID- 17427430 TI - [Blood cholesterol spectre in patients with acute and chronic inflammation of infectious origin]. AB - Low level of blood cholesterol is often found in patients with diseases which pathogenesis is mainly associated with inflamation. To detect blood cholesterol spectre, 383 patients with acute and chronic infections have been observed, level of blood cholesterol of 1259 patients with different pathology was retrospectively analyzed. It was found that an increase in frequency of low cholesterol and decrease in frequency of high cholesterol in patients with diseases not associated with infections do not depend on the age of patients. Extremely low level of cholesterol (Cholesterol < or = 100 mg/dl) is found in 12,8% of patients with inflamation of infectious origion, oftener in patients with community-acquired pneumonia and chronic virus hepatitis. Patients with intestinal infections have extremely low level of cholesterol; two-fold oftener than healthy persons have. PMID- 17427431 TI - [Substantation of usefulness of the combined sorption therapy in women in childbirth who have high risk of septic complications]. AB - Features of immune and cytokine status of women in childbirth who had high risk of septic complications and were under combined enteral and application sorption therapy with SILLARD-II have been studied. A new method of prophylaxis of septic complications in women who underwent Cesarean section and had a high risk of septic complications have been developed and introduced into practice. The author has showed that this proposed method is effective based on immune and cytokine status of women in childbirth. PMID- 17427432 TI - [Effect of cerebrokurin and lipin on changes in the rat brain mitochondria in experimental model of hypoxia]. AB - The authors carried out the sudy on morphofunctional changes of mitochondria of neurocytes of the brainstem of rats under experimental model of hypoxia and administration of Cerebrokurin and lipin. Cerebrokurin administration to rats which underwent severe chronic hypoxia lead to increase in mitochondria diameter and considerable increase of their area in the unit of volume as well it resulted in renewal of myelin sheaths. Use of Cerebrokurin and lipin in animals which underwent moderate hypoxia lead to probable increase in mitochondria area in the unit of volume in comparison with animals not treated with these agents. PMID- 17427433 TI - [Effect of plant biocomposites based on Georgian tea "per se" and in combination with cisplatin on Walker carcinosarcoma W-256 and Guerin's carcinoma growth rate in rats]. AB - Green tea biocomposite had effectivey hampered the growth of rat Walker W-256 carcinoma and in less extent rat Guerin's carcinoma. Black tea biocomposite had not practically influenced on Guerin's carcinoma growth. The biocomposite from green tea and extract from red vine rind and lemon suppressed at the level of tendency the growth of rat Walker W-256 carcinoma. The biocomposite from green tea and extract from red vine rind had hampered only Guerin's carcinoma growth and at the tendency had increased the growth of W-256 carcinosarcoma growth. This biocomposite increased also considerably the therapeutic efficiency of cisplatin on Guerin's carcinoma. Studed vegetable biocomposites posesses antiinflammatory and antioxidant properties. PMID- 17427434 TI - [The cytokine profile during experimental esophagitis and modification of cytoprotective mechanisms]. AB - The cytokine profile during experimental esophagitis and modification of NO-NOS and prostaglandin activity and the melatonin administration has been investigated. PMID- 17427435 TI - [Specificity of the training of modern manager for the system of Ministry of Health of Ukraine]. AB - The article devotes to the analyze of some questions of the training of managers for the system of Ministry of Health of Ukraine. PMID- 17427436 TI - [Stages of the development of military stomatology and oral surgery in main Clinical Military Hospital of Ministry of Defence of Ukraine]. AB - The article presents stages of the foundation and development of military stomatology and oral surgery in main clinical military hospital of ministry of defence of Ukraine for last 250 years. The authors outlined the role of scientists and physicians of the hospital in the treatment and prophylactics of teeth diseases of military persons as well as in studing dentofacial wounds and their replacement. Cumulative expirience was shown to be important for the development of military stomatology in total. PMID- 17427437 TI - [Contribution of Academician D. K. Zabolotnyi to the development of venereology (to 140th aniversary of birthday and 100th aniversary of opening of the first Syphilology Laboratory in Russia)]. AB - The Contribution of D. K. Zabolotnogo to the development of venerology is discussed in the article. Numerous scientific papers of D. K. Zabolotnogo are dedicated to aetiology, pathogenesis, epidemiology and prophylactics of three infections: pestilence, cholera and syphilis. PMID- 17427438 TI - An integrated TEAM approach to renal care. PMID- 17427439 TI - Nutritional supplements to be covered in Pa. by Chronic Renal Disease Program. PMID- 17427440 TI - Why measure quality in kidney disease? PMID- 17427441 TI - Helping helpers: a living donor mentor program. PMID- 17427442 TI - Expanded criteria donor kidneys: evolution and current practice. AB - Kidney transplantation is a well-established therapy for chronic renal failure, but its application is limited primarily by the availability of transplantable organs. The number of wait-listed patients continues to grow, and aggressive attempts to increase the numbers of transplants performed have failed to keep pace with demand. The continuing disparity between the demand for kidney transplants and the supply of organs has made efficient use of organs critical, forcing the transplant community to use organs from higher-risk donors than would previously have been considered. The precise definition of some marginal kidneys as expanded criteria donor kidneys was based on objective parameters and intended to expand the donor pool. This article reviews the development and definition of expanded criteria donor kidneys and the current strategies and outcomes associated with their use. PMID- 17427443 TI - Going the distance ... with renal disease. Part 3. PMID- 17427444 TI - Finding 'hope' for understanding kidney disease. PMID- 17427445 TI - DOPPS estimate of patient life years attributable to modifiable hemodialysis practices in Canada. AB - We examined data from the Canadian Organ Replacement Registry, and from a special substudy of CORR, to determine whether changes have occurred in practice patterns before and after the 1999 Canadian Society of Nephrology guidelines were published. Second, we used data from the Dialysis Outcomes and Practice Patterns Study to calculate the impact of observed deviations from guideline targets and estimated potential gains in life years that might accrue if guideline targets were achieved in all Canadian hemodialysis patients. For dialysis dose and hemoglobin targets, there was a significant improvement in Canadian facility performance over time. On the other hand, vascular access care showed a worse pattern with increased catheter use. A calculation of attributable risk, which assumes causality, suggests that 49 percent of deaths could be averted if all patients currently outside the guidelines achieved them over the next five years. When expressed as an annual death rate per hundred patient years, this corresponds to a decrease from 18 to 10.1 deaths per 100 patient years. We conclude that promoting a facility-based culture of quality improvement based on achievement of guideline targets is supported by international and Canadian observational data from the DOPPS. In the future, the impact of such an approach should be assessed empirically by correlating changes in practice over time with changes in outcomes. PMID- 17427446 TI - PCT network attacks 'unfair' reporting of top-slice effect. PMID- 17427447 TI - NHS Together. United on the streets. PMID- 17427449 TI - Time to tear ourselves away from paper. PMID- 17427448 TI - Primary care chiefs: reorganisation too far. PMID- 17427450 TI - The ticking clock. PMID- 17427451 TI - Paths to enlightenment. Pioneer providers. PMID- 17427452 TI - Commissioning. Line up to deliver. PMID- 17427453 TI - The way out west. PMID- 17427454 TI - Data briefing. Understanding preventable injury. PMID- 17427455 TI - Subtainable development. Cooking up a storm. AB - As one of the world's largest organisations, the NHS should make sustainable development a top priority. With a target-driven culture and ongoing financial pressure, sustainable development tends to struggle for a place on the agenda of senior NHS managers. The DoH has signalled its intensions to push trusts on sustainable development, and it may form part of future Healthcare Commission assessments. PMID- 17427456 TI - Service redesign. Backs for the future. PMID- 17427457 TI - Staff involvement. Life after Redfern. PMID- 17427458 TI - Sen. John Breaux: advising on healthcare policy. PMID- 17427459 TI - Changing the tax code for health insurance. PMID- 17427460 TI - 'The miracle on Meadows'. PMID- 17427461 TI - Banking on technology. AB - New technology is being used by banks to help providers improve their revenue cycle through: Digital lockbox service. Remote deposit. Web-based payment. Automated payment processing at the point of service. PMID- 17427462 TI - Effectively managing self-pay balances using KPIs. AB - Key performance indicators can be used to effectively monitor and improve self pay processes. Two keys to success in using KPIs to identify areas for improvement are: Begin with a limited number of KPIs. Commit to refining the measurements as you modify and improve your processes. PMID- 17427463 TI - The opportunity of price transparency. PMID- 17427464 TI - Dollars and sense engaging physicians in supply-cost control. AB - Five strategies for engaging physicians in supply-cost savings initiatives are to: Build a compelling case. Help physicians understand the cost profiles of the products they are using and their impact on cost of care. Give physicians a voice in supply-cost negotiations. Stick with the decisions you make in regard to vendors. Share the cost savings with physicians through investments in new equipment and technology. PMID- 17427465 TI - Trends in hospitals' use of contract labor. AB - During the period of 1997-2005, total personnel expense for the nation's short term hospitals averaged about 51.1 percent of total operating revenue. Contract labor expense as a percentage of total personnel expense increased from 1.3 percent to more than 3 percent during this period. By making more judicious use of contract labor, short-term acute care hospitals nationwide could reduce their staffing costs by as much as $3.5 billion. PMID- 17427466 TI - 7 steps toward gaining control of your labor costs. AB - The following steps can help you address your organization's labor costs: Ask critical questions. Create pay codes to track different additional labor expenses. Look for patterns in staffing. Understand your recruitment and turnover costs. Redefine bonuses. Review your overtime policies. Communicate. PMID- 17427467 TI - The value of vendor-reported ambulatory EHR benefits data. AB - Implementation of an electronic health record is expensive and labor-intensive. For this reason, providers often seek information about possible benefits to help them decide whether to implement an EHR. Our study found that a benefits database maintained by an ambulatory clinical systems vendor provided information that is useful, but that also has limitations. PMID- 17427468 TI - Just ASC: how to formulate the question. AB - When a hospital is considering whether to expand or transform its approach to outpatient surgery business, its internal financial and operating reports can help clarify trends and provide insight into the performance of the existing business over time. Benchmarking data can allow a hospital to compare its estimated performance for a proposed ambulatory surgery center with the actual performance of comparable existing facilities. Financial leaders need to be able to combine internal and external data into a pro forma to help determine whether to move ahead with plans to developthe ASC. PMID- 17427469 TI - "Joint" agreements: making the most of orthopedic contracts. AB - There are several steps your organization can take to lower orthopedic costs: Partner with physicians. Create incentives for physician involvement. Establish a technology review committee. Develop solicitation protocols. Leverage your group purchasing organization's contract portfolio. Negotiate discount agreements. PMID- 17427470 TI - The folly of using RCCs and RVUs for intermediate product costing. AB - Two measures for computing the cost of intermediate projects--a ratio of cost to charges and relative value units--are highly flawed and can have serious financial implications for the hospitals that use them. Full-cost accounting, using the principles of activity-based costing, enables hospitals to measure their costs more accurately, both for competitive bidding purposes and to manage them more effectively. PMID- 17427471 TI - Nurse staffing report cards. PMID- 17427472 TI - How big is your toolbox? PMID- 17427473 TI - Financial incentives for healthcare IT adoption. AB - For the pace of healthcare IT adoption to be accelerated, barriers such as the misalignment of financial incentives must be corrected. While adoption incentives are still in the formative stage, it is clear that mature incentives need to be sufficiently large to lead to adoption, emphasize outcomes rather than strictly use of IT, and require that hospitals put "skin in the game". Failure to develop these incentives will guarantee that the country will continue to experience low healthcare IT adoption rates. A collaborative effort by purchasers, providers, and government is the key to addressing the challenge of misaligned incentives. PMID- 17427474 TI - What would you do? What should this hosipital do about its physicians developing a freestanding endoscopy unit? PMID- 17427475 TI - Will CMS's adoption of severity-adjusted DRGs promote upcoding? PMID- 17427476 TI - Accurate documentation can improve your hospital's bottom line. PMID- 17427477 TI - Communication key to improved documentation. PMID- 17427478 TI - A few words may affect case mix, reimbursement. PMID- 17427479 TI - CMs, social workers collaborate on difficult cases. PMID- 17427480 TI - QI program reduces length of stay after hysterectomy. PMID- 17427481 TI - Hospital CoP changes affect EMTALA compliance. PMID- 17427482 TI - Administrators fall short of full recognition of CM duties. PMID- 17427483 TI - It takes all CM functions to provide necessary care. PMID- 17427484 TI - Patient safety alert. Integrated community effort wins Codman for FL hospital. PMID- 17427485 TI - [Incidentally discovered adrenal masses in the Department of Internal Medicine, Endocrinology and Hemostatic Disorders, Medical University of Gdansk]. AB - Clinically silent adrenal masses (incidentaloma) are incidentally discovered lesions when noninvasive imaging methods (ultrasonography--USG, computer tomography--CT, magnetic resonance imaging--MRI) are performed for the reason other than known or suspected adrenal disease. Most of studies report the prevalence of adrenal incidentaloma range between 1 and 10% in radiological series. MATERIAL AND METHODS: Between 1993 and 03.2004 we observed 198 patients with incidentalomas of adrenal glands (144 females--72.7% and 54 males--27.3%). RESULTS: After endocrinological evaluation, 164 patients were qualified for surgery. In 119 (72.5%) cases open adrenalectomy was performed, and in 45 (27.5%) laparoscopic adrenalectomy was done. Adrenocortical adenoma was diagnosed in 54.9%, adrenal hyperplasia in 8.5%, adrenal carcinoma in 6.7%, pheochromocytoma in 12.9% (in 3.1% of all cases malignant pheochromocytoma was diagnosed), in 4.9% others malignant tumors (primary or metastatic), in 4.9% adrenal cysts and in 7.1% other rare adrenal pathologies were found. CONCLUSION: All malignancies were found in tumors with the diameter over 3 cm. In tumors with diameter over 6 cm malignant cases were found in 70.8%. PMID- 17427486 TI - [SEN virus infection in maintenance hemodialysis patients]. AB - SEN virus (SENV) is a new, mostly parenterally transmitted, hepatotropic agent. The prevalence of SENV among patients undergoing maintenance hemodialysis (HD) in Poland, as well as risk factors for the infection are not established. Serum samples of 91 patients receiving maintenance HD in Bialystok were tested for the presence of strain H SENV (SENV-H) DNA by means of polymerase chain reaction. Fifty-one non-dialysis subjects, age- and sex-matched with the HD patients, mostly with chronic kidney diseases (96%), without hepatitis B (HBV) and C (HCV) or the history of blood transfusion and donation served as controls. SENV-H viremia was prevalent in 40% of HD patients and in 2% of control subjects (p < 0.0001). On multivariable logistic analysis, neither age (63.1 +/- 13.2 years), gender (49% females), dialysis vintage (29, 2-200, months), previous transfusions of packed red blood cells (84%) or fresh frozen plasma (4%), seropositivity for HBs antigen (13%), HCV antibodies (23%) or HCV RNA (17%) were independently associated with SENV-H prevalence in HD patients (chi2 for the model = 13.3, p = 0.103). No associations between SENV-H status and clinical or biochemical markers of liver disease, including serum aminotransferases levels were observed. In conclusion, SENV-H viremia is widespread among patients receiving maintenance HD in north-eastern Poland. Risk factors for its occurrence are equivocal; the infection may well be transmitted by parenteral and feco-oral routes. SEN virus is not directly responsible for liver damage in maintenance HD patients. PMID- 17427487 TI - [The influence of genetic polymorphism of CYP2E1 on the development of alcohol liver cirrhosis]. AB - Alcoholism is a significant medical, social, and economic problem. Genetic polymorphism of enzymes involved in alcohol metabolism plays a relevant role in etiopathogenesis of alcohol disease and alcohol liver cirrhosis. The aim of the study was the evaluation of the influence of genetic polymorphism of CYP2E1 on the development of the alcohol abuse and alcohol liver cirrhosis the Polish population. MATERIAL AND METHODS: The CYP2E1 genotype and c1 and c2 alleles frequency were examined in 188 patients. Genotyping of the CYP2E1 was performed using polymerase chain reaction-restriction fragment length polymorphism method on white cell DNA. RESULTS: In the examined population encompassing 188 subjects the c2 allele was present only in 1.06% of patients. It was found only in patients abusing alcohol. In the group of patients with alcoholic cirrhosis it was present in 3.5% of cases. The c1/c2 genotype was present in 2.12% of subjects. The c2/c2 genotype was not found in any patient. Heterozygotes cl/c2 were present only in 7% of patients with alcohol liver cirrhosis. The c2 allele and cl/c2 genotype occurred statistically significantly more frequently in patients with alcohol cirrhosis than in control group. Patients possessing the c2 allele and cl/c2 genotype statistically significantly earlier initiated the alcohol abusing than those in which the c1 allele and c1/c1 genotype were present. CONCLUSION: Our studies suggest that the frequency of allele c2 in Polish population is low, but the presence of c2 allele may be a risk factor for the alcohol liver cirrhosis. PMID- 17427488 TI - [Activity of antioxidant enzymes in patients with liver cirrhosis]. AB - The aim of our studies was the estimation of activities of antioxidant enzymes in patients with liver cirrhosis. We investigated activities of superoxide dismutases (CuZnSOD, MnSOD), catalase (CAT), selenium dependent GSH peroxidase (Se-GSH-Px), selenium independent GSH peroxidase (non-Se-GSH-Px), GSH-S transferase (GST), GSH reductase (GSHR) and the level ofreduced gutathione (GSH) in cirrhotic and healthy liver tissues. The activities of CuZnSOD, MnSOD, CAT and GSH-dependent enzymes (except GSHR) were found to be lower in cirrhotic tissue compared to healthy liver. Those changes were associated with decrease of GSH level in cirrhotic tissue compared with control liver tissue. Our results show that antioxidant barrier in liver cirrhosis is impaired. It is associated with decrease of glutathione level and changes of activities of antioxidant enzymes (SOD, CAT, GSHPx, GST, GSHR) in liver cirrhosis compared with healthy liver. PMID- 17427489 TI - [Eventrations--the prospective analysis of risk factors of eventrations patients after laparotomy treated in 8 surgical departments of Podkarpacie District]. AB - Eventration is still a major surgical problem. Although not very common, but may cause serious complications leading to patient's death. It is observed in 0.2-7% (average 2%) of all the laparotomy. The aim of the study was the analysis of risk factors and frequency of eventrations. MATERIAL AND METHODS: The study was performed in 8 surgical departments in Podkarpacie district since April 2004 till March 2005. RESULTS: During the analysed period there were 4360 laparotomies performed. Eventration was observed in 23 cases (0.52% of all laparotomies performed)--30.5% of the afflicted were females; males constituted the remaining 69.5%. Patients over the age of 70 represented approximately 70% of all of the eventration cases. The most common risk factors were: ASA (American Society of Anesthesiologists) III or higher, low plasma protein level, operations on the large bowel, abdominal midline incision and emergency operations. Occurring eventrations were aseptic, late, III degree. CONCLUSIONS: (1) The frequency of eventrations in hospitals of podkarpacie district is low (0.52% of all laparotomies indicates good technique of wound suture after laparotomy). (2) The eventrations were associated with the following risk factors: age over 70, ASA greater or equal III, low plasma protein level, operations on the large intestine, midline incision and emergency surgeries. PMID- 17427490 TI - Quality of life before and after laparoscopic fundoplication. Does quality of life depend on psychological factors? Preliminary report. AB - Gastroesophageal reflux disease (GERD) is the most common benign foregut disorder. In many cases multiple factors contribute to the development of esophagitis and result in GERD symptoms. GERD has a definite negative impact on patients' quality of life. The aim of this study was to assess the postoperative results and compare pre- and postoperative quality of life after laparoscopic Nissen or Toupet fundoplication and also to determine if psychological factors have an influence on the quality of life and surgical outcome. MATERIAL AND METHODS: Fifty patients with a proven GERD and hiatal esophageal hernia (HHE) were included in a prospective study. After routine examinations, assessment of quality of life (Gastrointestinal Quality of Life Index--GIQLI) and psychological investigation (Minnesota Multiphasic Personality Inventory--MMPI) the patients underwent laparoscopic fundoplication. Three months after operation all of the patients had quality of life assessment again. RESULTS: Patients were divided into two groups depending on the result of MMPI test. Patients with conversion disorder (CD) preoperatively had significantly lower mean overall score in GIQLI (84.3 +/- 16.3) in comparison to patients without CD (93.0 +/- 19.4). The mean score increased significantly in both groups 3 month after surgery (to 100.6 +/- 21.2 in patients with CD and to 106.7 +/- 18.1 in patients without CD). CONCLUSION: Quality of life in GERD patients is significantly impaired. It shows general improvement after surgery. There are findings in the literature and in our research project showing that comorbidities or psychological factors are able to affect the subjective outcome without any fundamental explanations or basis. Thus, psychological intervention can decrease the spectrum of symptoms and improve the subjective outcome of fundoplication in selected patients. PMID- 17427491 TI - [Taste sensitivity to natrium chloride and saccharose in women and men of different age]. AB - The gender and age influence on saccharose taste and sodium chloride taste sensitivity was tested in this work. MATERIAL AND METHODS: 275 volunteers, i.e. 161 women and 114 men, aged 18-55, participated in the study. All individuals were healthy and non-smoking. Serials dilutions of saccharose and sodium chloride were used to analyze sweet and salt taste sensitivity. The examination was performed according to the procedures established by the Polish Committee of Standards (PN-ISO 3972: 1998). The findings suggest no link between the gender and saccharose and sodium chloride taste sensitivity. However there is a link between taste sensitivity and age, i.e. taste sensitivity to sodium chloride decreases significantly whereas taste sensitivity to saccharose decreases slightly with age ofpopulation. PMID- 17427492 TI - [Metastases of the melanoma to the intestine]. AB - The cases of 4 patients with primary skin melanoma, who underwent resection of the small intestine due to the intestinal metastases, are presented. Two patients died as a result of metastases after intestinal resection; the other two have been alive for a year and 10 years without any symptoms of the disease. The presence of the melanoma metastases to the intestine can be suspected in patients with anaemia caused by bleeding into the lumen of an intestine, ileus, rapid weight loss or noticeable tumor in the abdomen. The resection of intestine with metastases should be chosen as a safe method, which allows good palliation; it also offers the chance for a longer life. PMID- 17427493 TI - [Clinical characteristics of 33 patients with adrenal incidentaloma]. AB - Incidentally diagnosed, clinically asymptomatic adrenal masses (incidentaloma) have become more and more common problem in everyday clinical practice. The prevalence of such tumors is 2.3% in autopsy series and 0.5-2% in computed tomography series. The aim of the study was a clinical analysis of patients with adrenal incidentaloma, hospitalized in the Department of Internal and Metabolic Diseases of the Medical University of Silesia. MATERIAL AND METHODS: 33 patients, including 25 females (75.7%) and 8 males (24.3%), were investigated. The mean age was 59.0 +/- 10.7 year (31-80) and the mean body mass index (BMI) was 29.6 +/- 5.9 kg/m2 (19.4-47). RESULTS: 16 patients (48.5%) were obese and 10 more patients (30.3%) were overweight. Hypertension was diagnosed in 25 patients (75.8%), diabetes or impaired glucose tolerance in 9 patients (27.3%) and hyperlipidemia in 20 patients (60.6%). Endocrine tests revealed: disturbed cortisol rhythm in 1 patient, uncertain result of the overnight dexamethasone suppression test in 4 patients, elevated urinary 17-hydroxycorticosteroids excretion in 7 patients, elevated urinary 17-ketosteroids excretion in 2 patients and increased urinary excretion ofvanillylmandelic acid in 2 patients. The tumor was located in the right adrenal gland in 13 patients (39.4%) and in the left adrenal gland in 16 patients (48.5%). 4 patients (12.1%) demonstrated bilateral adrenal masses. At least one of the tumor sizes was greater than 40 mm in 3 patients (9.1%). CONCLUSION: Clinically asymptomatic adrenal tumors occurred more frequently in overweight or obese women, between 51-70 years old, with lipid disorders and hypertension. PMID- 17427494 TI - [Malignant neoplasm of nose and paranasal sinuses in Lower Silesia in years 1992 2001]. AB - Malignant neoplasms of the nose and paranasal sinuses occur rarely but due to late diagnosis and poor treatment effects still remain a serious problem. The aim of the study was the analysis of all nose and paranasal sinus neoplasms treated at Lower Silesia in the years 1992-2001. MATERIAL AND METHODS: In the years 1992 2001 in Lower Silesia region there were 182 patients treated for malignant nose and paranasal sinus tumors. Males outnumbered females significantly with male : female ratio was 1.9:1. Principal management was combined therapy--surgery with radiotherapy in 84% of the cases. Radiotherapy alone was performed in 8.8% and chemotherapy as palliative treatment in 7.1%. Fifteen patients (8.2%) underwent radical neck dissection followed by radiotherapy. RESULTS: The analysed group constituted 79.6% patients with epithelial tumors and 20.4% with nonepithelial tumors. Cases T3-T4 predominated in the material in amount of 77.9% (113). There were 15 T2 cases (10.4%) and none of T1. Most of the tumors (57%) arose from middle segment of ethmomaxillary sinuses. Upper segment tumors constituted 18.6% of cases and lower segment--10.4%. Enlarged lymph nodes were diagnosed in 8.2%. Survival rate analysis in patient treated in the years 1992-2001 revealed 37.9% (69) of 3-year survival and 30.2% (55) of 5-year survival. CONCLUSIONS: Unsatisfactory results of treatment are an effect of a high advanced stage of the tumor while diagnosed. Its direct causes are: asymptomatic course of the disease, localization in complex anatomical structure, not enough experience among medical staff dealing with the disease very rarely. PMID- 17427495 TI - [The evaluation of the brain arteriovenous malformations' operative treatment in the clinical material of Silesian Medical University's Departament of Neurosurgery]. AB - A risk of haemorrhage in arteriovenous malformations (AVM) of the brain is estimated as 2-4% per year. A mortality rate from this reason is estimated as 1% per year and morbidity 10-20%. The methods of treatment ofAVM are: microsurgical treatment, radiosurgery, endovascular treatment. The aim of this work is presentation of our results of surgical treatment of AVM and comparison with the results of radiosurgical and endovascular treatment presented by other authors in the literature. MATERIAL AND METHODS: Between 1990-2002 in the Department of Neurosurgery of Medical University of Silesia in Katowice 31 patients were operated with AVM's of the brain. Among them there were 10 female and 21 male at the age from 10 to 69 years. The average age was 36.1. The first sign was intracranial haemorrhage in 19 cases and epileptic seizures in 9 cases. In all cases the cerebral angiography was performed and all patients were assessed as I to III score according to the Spetzler-Martin scale. All patients were operated on using microneurosurgical techniques. The total removal of the tumour was assessed using the intraoperative Doppler examination. The state of the patients at the discharge was estimated according to Glasgow Outcome Scale (GOS). RESULTS: In all cases the malformations were removed totally. In 2 cases (6.4%) we noticed the deterioration of neurological condition after operation. The state of 28 patients (90.3%) was assessed as very good and good (I or II score) according to GOS at the discharge. We didn't notice any mortality in our group of patients. CONCLUSIONS: In the conclusion we want to emphasize the advantages of surgical treatment of AVM's of the brain especially these including in I to III score according to Spetzler-Martin scale comparing with the results ofradiosurgical and endovascular methods of treatment. PMID- 17427496 TI - [The role of brain magnetic resonance studies in the diagnostics of central nervous system lesions in HIV-1 positive patients]. AB - Approximately 10% of persons infected with human immunodeficiency virus (HIV) and 75-90% patients with acquired immunodeficiency syndrome (AIDS) present neurological symptoms. This group causes the greatest diagnostic difficulties among AIDS patients. The purpose of the study was to demonstrate the usefulness of the magnetic resonance imaging (MR) of the central nervous system (CNS) in the diagnosis and differentiation of primary and secondary lesions. MATERIAL AND METHODS: Fifty one patients with the symptoms suggesting CNS involvement were enrolled into the study. In 40 of them AIDS diagnosis was established based on the picture of CNS lesions. MR studies were performed using EDGE-ECLIPSE 1.5 T unit with FSE sequences, T1- and T2-weighted images, before and after paramagnetic contrast agent injection, in sagittal, coronal and transverse planes. In almost all cases FLAIR technique was also used. Thirteen persons were examined several times. Altogether we executed 62 studies. Magnetic resonance results were compared with the results of laboratory tests and sometimes histopathological examination at autopsy. RESULTS: Pathological findings were visible on MR of CNS in 37 (92.5%) of 40 patients with AIDS. In 8 (20%) cases the diagnosis of AIDS dementia complex (ADC) was established. Toxoplasmosis was diagnosed in 9 (22.5%) cases, tuberculosis in 6 (15%), cryptococcosis in 5 (12.5%) patients, progressive multifocal leukoencephalopathy (PML) in 3 (7.5%) patients, primary lymphoma also in 3 (7.5%), and atypical mycobacteriosis in 2 patient. In 1 (2.5%) case we could not establish the final diagnosis. We described the patterns of MR appearance in the different types of lesions mentioned above based on our findings and the literature review. CONCLUSIONS: Magnetic resonance imaging has a high sensitivity in identifying lesions of the central nervous system. It is very useful, together with the results of other tests, in the final diagnosis of the lesions in the CNS. Magnetic resonance plays also an important role in the treatment planning and monitoring of the efficacy of therapy used in AIDS patients. PMID- 17427497 TI - [Port device central venous access in children with chronic renal disease- personal experience]. AB - The application of central venous lines in children has been widely accepted in the case of pediatric cancer treatment. This is of particular importance when the treatment must be continued during the long period of time. The indication to long-term application of central venous lines became significantly frequent within last years. They are necessary in the treatment of chronic pediatric patients, in whom the central venous line allows continuous access for medication, parenteral rehydration, nutrition and frequent blood sampling. MATERIAL AND METHODS: In the current study authors present their experience in subcutaneous port devices application in children with kidney disease. The case history data obtained from 8 children were retrospectively analysed. In these children subcutaneous port devices were applied for mean 26.7 months (totally 9 port devices). The mean age at the time of implantation was 2.2 years, and the mean body weight--10.6 kg. RESULTS: Peripheral venous access in all children was bad. In one child during the time of implantation the hematoma of coli and chest was present. Infectious complications connected with implanted port device were not detected. Thrombotic complications were present in 6 children with chronic renal failure--in 5 the lumen of port device has been successfully recanalysed, in 3 cases even several times. In 1 child the thrombus on the tip of central venous line was detected. In 2 children the removal of port device was necessary because of breakage of venous line and in the second case because of port device thrombosis. Two children died with functioning port device. The cause of death was not connected with implanted port device. CONCLUSIONS: The application of subcutaneous port devices definitely improved the comfort of treatment but was significantly associated with thrombotic complications. Infectious complications were not detected as compared to hematological group of patients. PMID- 17427498 TI - [Assessment of postvaccinal response in children past pertussis]. AB - In the nineties, despite high percentage of vaccination of children, from 86% to 99% dependently on the region, there came to the increase of whooping cough cases. Until then infants and children to the age of 5 years suffered from this disease, whereas in the last decade the number of cases increased mainly among children over 5 years of age, who were subjected to full cycle of vaccination against whooping cough. Searching for the causes of such epidemiologic situation the following suggestions have been given: change of the bacteria antigenicity, the phenomenon of postvaccinal immunity extinction, immune system mechanisms disorders and groundless excuse from vaccination. The study estimates parameters of postvaccinal response to immunization with selected vaccines in children post whooping cough. PMID- 17427499 TI - [Surgical treatment of Parkinson's disease as a supplement to pharmacological therapy]. AB - The aim of the study was review of literature and description of surgical treatment of Parkinson's disease, based on neuroanatomical and neurophysiological aspects ofnigrostriatal system. Parkinson's disease is the neurodegenerative disorder characterized by dopaminergic cell loss in the compact part of substantia nigra. It is treated pharmacologically or surgically. The surgical treatment is the supplement to the pharmacological one in severe and drug resistant cases. In this study the common methods of surgical treatment of Parkinson's disease: ablative lesions and deep brain stimulation procedures in globus pallidus internus, thalamus and subthalamic nucleus and also the intrastriatal transplantation ofdopaminergic cells (into the striatum) were described. PMID- 17427500 TI - [Oxytetracycline--mechanism of action and application in skin diseases]. AB - Oxytetracycline is a bacteriostatic antibiotic. Newly discovered, additional mechanisms of action include antioxidant, antiinflammatory and immynosupresive activity of oxytetracycline and other tetracyclines. These activities were the basis for developing therapy regimens with oxytetracycline in subantimicrobial doses. Due to its significant efficacy, limited adverse effects and low therapy costs, oxytetracycline at the dose of 500 mg per day is presently considered as therapy of choice in papulopustulous acne. Rosacea and perioral dermatitis are other indications. Topical oxytetracycline shows significant efficacy in primary and secondary skin infections with inflammatory reaction. PMID- 17427501 TI - [Fatty liver disease--mechanism, clinical significance and factors with a special regard to primary hepatotropic viruses]. AB - Fatty liver disease is one of most frequently diagnosed hepatopathies while detailed examination of potential causes of liver enzymes abnormalities is done. Despite its isolated nosology fatty liver disease often co-exists with other liver pathologies or - more often - it is their natural consequence. Liver steatosis is also more often found in primary hepatotropic viral infections. However, it's more prevalent among HCV (hepatitis C virus) infected persons than in those HBV (hepatitis B virus) infected. It is described in 30-70% routinely pursued liver biopsies in HCV infected individuals. Hitherto, there were many analyses concerning clinical and prognostic implications pursued of HCV influence upon inflammatory and fibrotic process of the liver. This article is an up-to date review of current clinical and therapeutic implications of hepatosteatosis supported by referent study results. PMID- 17427502 TI - [The guidelines for the effective prevention of pressure ulcers]. AB - The pressure ulcers belong to hard healing chronic wounds which can be the cause of heavy systemic complications delaying recovery to health and exposing patients to psychophysical suffering. They increase the demand for medical care and cause the growth of general costs of treatment and hospitalization. The most subjected to the development of bedsores are long-term immobilized and severely ill patients. Therefore, identification of high risk patients and the initiation of preventive workings are highly recommended. The aim of this work was to indicate the essential elements of prophylaxis of bedsores that are helpful in organization ofnurses' work. They include the following elements: risk estimation, decreased pressure and mechanical forces, correct state of skin, pain control, assurance of proper nutrition state, patient's activation, complex and individualized care. To achieve high effectiveness of prevention the teams initiating and estimating the effectiveness of standards and prophylaxis programs and treatment of chronic wounds are appointed. The aim of their work is to reduce the prevalence and the development of bedsores among menaced patients. PMID- 17427503 TI - [Polycystic ovary syndrome and hair unit function disturbances in dermatological practice]. AB - Polycystic ovary syndrome (PCOS) is the common endocrine disorder of reproductive age women which is characterized by hyperandrogenism, chronic anovulation, and increased risk for infertility, endometrial cancer, developing metabolic dysfunction (type II diabetes, dyslipidemia), hypertension and heart disease. The syndrome is also associated with some skin disorders: hirsutismus, alopecia androgenetica and acne. The successful dermatologic therapy requires the holistic diagnosis of the women with skin disorders described above and use the antiandrogenic treatment with conventional methods and laser depilation in hirsutismus. In this paper we present literature studies and our own experiences. PMID- 17427504 TI - [Potential prognostic value of angiogenesis, cell proliferation and metastasing in patients with surgically treated gastric cancer--current knowledge]. AB - Gastric cancer is nowadays one of the most severe human malignant tumours. Surgeons and oncologists often encounter in their clinical practice the course of disease that cannot be fully accounted for by commonly used classical histo clinical prognostic parameters. Gastric tumours of similar size, similar progression stage and similarly treated may develop along significantly different courses, the phenomenon that seems to depend on as yet not fully recognized biological factors. Preliminary observations from different centers concerning the role of new molecular parameters having potential influence on prognosis in gastric cancer are often contradictory. This study based on bibliography and own observations presents current data connected with potential prognostic value of angiogenesis, proliferation and metastasing in patients with surgically treated gastric cancer. It is very possible that in the future the evaluation of these molecular parameters will find its place in clinical practice for determination of more precise prognosis and improving distant results of combined treatment in gastric cancer patients. PMID- 17427505 TI - [Vasovagal syncope--diagnostic difficulties in adolescents]. AB - Consciousness disturbances in adolescents may be caused by different reasons. Among them, cardiologic and neurological backgrounds should be considered. The most important diagnostic tests the vasovagal syncopes are: tilt-table test, brain oxygenation measurement, routine and extended elektroencephalography. Performing of these examinations simultaneously may help in determining cause of symptoms. Our patient serves us as an example. PMID- 17427506 TI - [Self-mutilation as a consequence of the perinatal brachial plexus palsy]. AB - In this study we presented 2 cases of the self-mutilation of the hand following obstetrical brachial plexus palsy. The mechanism of this behaviour was also discussed. PMID- 17427507 TI - [Hereditary angioedema in a 16-year-old girl]. AB - The angioedema may be connected with immunological or allergic reactions, rarely appears as a genetically determined hereditary disorder. The cause of hereditary genetically determined angioedema is the defect of complement due to lack or decreased activity of Cl esterase inhibitor with the low serum C4 complement. The acquired angioedema is the most frequently the effect of lymphoproliferative and autoimmunological diseases. Hereditary angioedema was clinically characterized by subcutaneous oedema of extremities, face, neck, throat, gastrointestinal tract and brain. Tranexamic acid, sigma-aminocaproic acid and anabolic steroids- Danazol and Stanazol are administered for short- and long-term prophylaxis. Life threatening recurrent episodes of angioedema were treated with replacement therapy. We present a case of hereditary angioedema in a 16-year-old girl admitted to hospital with massive facial oedema. The recurrence of symptoms without any effect of typical treatment was the indication for additional diagnostic tests. Diagnostic and therapeutic problems and a variable clinical course of disease were described. PMID- 17427508 TI - [Primary hypertrophic osteoarthropathy as a rare cause of joint pain]. AB - Primary hypertrophic osteoarthropathy (HOA) is a rare cause ofjoint pain. The diagnosis is often delayed, due to mild complaints. However, finger clubbing combined with joint pain should suggest considering HOA in the diagnosis. The progression of toes and fingertips widening is slow and can be overlooked by a patient. It is common that patient seeks physician's aid only when the joint pain occurs. The presented case is an example of delayed diagnosis. Due to the fact that secondary HOA coexists with some diseases associated with serious prognosis it is recommended to exclude other possible causes before diagnosing primary HOA. Some clues, like the age of disease onset or positive family history allow the diagnosis of primary HOA with high probability. This case report contains clinical symptoms and new diagnostic criteria, differential diagnosis, and possible complications. PMID- 17427509 TI - [Tuberculous peritonitis--rare cause of peritonitis]. AB - A case of 73-year-old male patient operated due to tuberculous peritonitis was presented. Diagnostic difficulties in the recognition of this rare site of tuberculosis were discussed. The role of explorative laparotomy and histopathologic evaluation of the material obtained during operation was emphasized. PMID- 17427510 TI - [Between obligation and duty--on deontological codes in medical professions]. AB - The social role of medical occupations is changing, along with their professional ethics. It could be argued that the role of ethical regulation (ethical codes) in these professions is diminishing. It could be due to ethical standards increasingly taking on character of legal norms. The notion of "moral obligation" is thus subject to transformation, as it progressively denotes legal duty, i.e., coercion. Legal validity of ethical standards is defined by lawyers. The continual tendency of occupational ethics to resemble rules of law may thus question the effect of ethical regulation in medical professions. The latter are performed not only because of official or economical compulsion but also because of moral obligation and the medic's desire to do well. The standing of occupational ethics in the work ofphysician, nurse and pharmacist is a prerequisite for maintaining the high social prestige of these professions. PMID- 17427511 TI - [Nitric oxide metabolism]. AB - Nitric oxide is an important mediator of physiological and pathological processes. It is a lipophilic molecule that contains a single unpaired electron which causes NO to be chemically reactive, and to function as a free radical with a short lifetime. NO can act by direct and indirect effects. Direct effects occur between NO and specific biological molecules whereas indirect effects are mediated by reactive nitrogen oxide species (RNOS) formed from the reaction of NO either with oxygen or superoxide. This review discusses the metabolic pathways of NO. PMID- 17427512 TI - [Comment to the article: Unique massive recurrence liposarcoma of the retroperitoneum. AFollow-Up Studies. Wiewiora M, Pardela M, Kobielski, published in Wiad Lek. 2006;59(7-8); 577-9]. PMID- 17427513 TI - Chemotherapy in patients with recurrent or refractory epithelial ovarian cancer. AB - OBJECTIVES: To evaluate the response rates, progression-free survival, and overall survival of patients with epithelial ovarian carcinoma who were treated with chemotherapy after being resistant to or had recurrence after first-line chemotherapy. MATERIAL AND METHOD: Clinical and pathological data of all patients with epithelial ovarian carcinoma who received chemotherapy in the second-setting at the Department of Obstetrics and Gynecology, Bangkok Metropolitan Administration Medical College and Vajira Hospital from January 1994 to December 2005 were reviewed. RESULTS: During the study period, 61 ovarian carcinoma patients met the inclusion criteria. All patients had primary surgery, not responded to or had recurrence after first-line chemotherapy, and received subsequent chemotherapy. Thirty-seven cases (60. 7%) were considered as platinum resistant and 24 cases as platinum-sensitive (39.3%). The overall response rate (RR) to subsequent chemotherapy was 23.0% (14 patients): complete response 18.0% (11 patients) and partial response 5.0% (three patients). Stable disease was achieved as the best response in 11 patients (18.0%). Thirty-six patients (59.0%) experienced disease progression. Median progression-free survival (PFS) of all 61 patients was 5.7 months (95%CI, 4.7-6.7 months) while median overall survival (OS) was 18.3 months (95%CI, 2.7-34.0 months). Some prognostic factors were studied and found that patients with platinum-sensitive had a better response rate, longer PFS and OS than those with platinum-resistant diseases. CONCLUSION: Response rate of ovarian carcinoma to subsequent chemotherapy for resistant or recurrent diseases was modest. Median PFS and OS of the patients were less than and slightly longer than a year respectively. The patients who had platinum sensitive diseases had a better prognosis in terms of RR, PFS, and OS than those with platinum-resistant disease. PMID- 17427514 TI - Comparison of osteoporosis self assessment tool for Asian (OSTA) and standard assessment in Menopause Clinic, Chiang Mai. AB - OBJECTIVE: To compare osteoporosis self assessment tool for Asian (OSTA) with the standard measurement of bone mineral density (BMD) by dual energy x-rays absorptiometry (DXA) in- identifying the prevalence of osteopenia and osteoporosis in Thai menopausal women who attended the Menopause Clinic, Faculty of Medicine, Chiang Mai University. MATERIAL AND METHOD: The data was retrospectively collected from the medical records of women who had lumbar femoral neck and radius BMD t-score measurement by DXA (Hologic, QDR-4500C) between January 2004 and December 2005. The body weight and age ofpatients were calculated for the OSTA index score. The women with OSTA score < -1 and > or = -1 were classified as "moderate to high risk" and "low risk" for fracture, respectively. The BMD T-scores of < -1 and > or = -1 were classified as "osteopenia to osteoporosis " and "normal" respectively. RESULTS: Three hundred and fifteen menopausal women of the age of 45-87 years were included. The BMD T scores assessment revealed that more than half of the subjects had osteopenia to osteoporosis (T-score < or = -1), 196 women (62%) at lumbar spine, 133 women (42.2%) at femur and a third, 114 women (36.2%) at radius respectively. The OSTA index at the standard cut-point of < -1 had a sensitivity and specificity of 36.2% and 71.4% respectively for the lumbar spine and 40.6% and 72.0% respectively for the neck of the femur in predicting osteopenia to osteoporosis status. CONCLUSION: The OSTA index score had a low sensitivity (36-48%) but a high specificity (71-75%) for identifying osteopenic to osteoporotic women among population of menopausal period Nevertheless, it is a useful screening tool in old age women (> 65 years). PMID- 17427515 TI - Effect of a new oral contraceptive with drospirenone on vital signs, complete blood count, glucose, electrolytes, renal, and liver function. AB - OBJECTIVE: To evaluate the effect of a new oral contraception formulation with drospirenone (Yasmin) on vital signs, complete blood count, glucose, electrolytes, and renal and liver function. MATERIAL AND METHOD: An open-label non-comparative clinical trial was conducted. One hundred women who were planning to use oral contraception for at least six months were recruited. The subjects received a blister pack which contained 21 tablets of 3 mg drospirenone /30 tg ethinyl estradiol for the first four cycles (1 cycle = 28 days). Cycle 5 and 6 blister packs were dispensed during the visit in cycle 4. Heart rate and blood pressure of each subject were checked at baseline and each visit. Serum from each subject was collected and sent for complete blood count, glucose, electrolytes, and renal and liver function tests at baseline and at cycle 6. Mean differences in these tests at cycle 6 compared to baseline were assessed. RESULTS: Ninety-two of the 100 subjects (92%) completed the present study. There was no significant change in heart rate, blood pressure, complete blood count, glucose, electrolytes, and renal and liver function tests at cycle 6 when compared to baseline. CONCLUSION: Oral contraception formulation with drospirenone (Yasmin) is well tolerated and has good contraceptive efficacy. It is safe, as it has no effect on heart rate, blood pressure, complete blood count, glucose, electrolytes, and renal and liver function. PMID- 17427516 TI - Incidence of large-for-gestational age newborn: a comparison between pregnant women with abnormal and normal screening test for gestational diabetes. AB - OBJECTIVE: To evaluate the incidence of large-for-gestational age (LGA) newborn between pregnant women with abnormal and normal glucose challenge test (GCT). DESIGN: Retrospective cohort study. MATERIAL AND METHOD: Two hundred and sixty pregnant women, who were at risk for gestational diabetes mellitus (GDM), received screening following practice guideline. The women were divided into two groups. The study group comprised of 130 women whose screening test results of 50 g 1-hour GCT were abnormal but had not been diagnosed with GDM (normal oral glucose tolerance test). The control group comprised of 130 women whose GCT results were normal. Comparison of various maternal and neonatal characteristics as well as the incidence of LGA between the groups was made. RESULTS: There were no significant differences in age, gestational age at first antenatal care, body mass index, and risk of GDM between the two groups. The study group had a significantly higher number of parity and number of risk factors of GDM than the control group. There was no significant difference in the incidence of LGA newborn between the two groups (8.5% in the study group and 10.8% in the control group, p = 0.528). There were also no significant differences in gestational age at delivery, pre-eclampsia, pre-term delivery, hyperbillirubinemia of the newborn between the two groups. There were no cases of maternal acute postpartum hemorrhage, and birth asphyxia. CONCLUSION: The incidence ofLGA newborn was similar between non-GDMwomen with abnormal and normal screening GCT results in Siriraj Hospital. PMID- 17427517 TI - Prevalence of bacterial vaginosis in Thai pregnant women with preterm labor in Siriraj hospital. AB - OBJECTIVE: To study the prevalence of bacterial vaginosis (BV) in pregnant women with preterm labor in Siriraj hospital. MATERIAL AND METHOD: A cross-sectional study of 158 pregnant women with suspected preterm labor was performed between January and July 2005. The subjects enrolled in the present study were between 28+0 and 36+6 menstrual weeks. BV blue test was performed on the vaginal fluid collected from lower one- third of vagina. RESULTS: The prevalence of BV in women in the preterm labor group was 25.8% compared to 14.1% in the preterm contraction group (p = 0.07). CONCLUSION: Compared with preterm contractions a higher prevalence of BV was found in the pregnant women with preterm labor. Given that a quarter of pregnant women with preterm labor tested positive for BV, it might be appropriate to perform this test in the triage setting. PMID- 17427518 TI - The association between meconium-stained amniotic fluid and chorioamnionitis or endometritis. AB - OBJECTIVE: Assess the association between meconium-stained amniotic fluid and chorioamnionitis or endometritis in term pregnant women. MATERIAL AND METHOD: A five-year retrospective study was undertaken between January 1, 1999 and December 31, 2003. One thousand seventy-nine pregnant women who delivered at the Department of Obstetrics & Gynecology, Phramongkutklao Hospital were included in the present study. RESULTS: Five hundred andfifty-three pregnant women (51.25%) had meconium-stained amniotic fluid (group 1) and 526 (48.75%) pregnant women were clear of amniotic fluid (group 2). Two pregnant woman in group 1 (0.36%) and eight pregnant women in group 2 (1.52%) were found to have chorioamnionitis (OR = 0.235). Postpartum endometritis was detected in only one (0.18%) pregnant women in group 1 and nine (1.71%) pregnant women in group 2 (OR = 0.104). CONCLUSION: No association was found between meconium-stained amniotic fluids and chorioamnionitis or endometritis. PMID- 17427519 TI - Influenza viral infection in 2005-2006 in Samitivej Hospital. AB - OBJECTIVE: Set an alarm system for early detection of respiratory viral infection. MATERIAL AND METHOD: The authors prospectively investigated avian flu and SARS between March 2005 and April 2006. Specimens from a nasopharyngeal swab or bronchial washing were analyzed for influenza A, B, parainfluenza, adenovirus, respiratory syncytial virus and avian flu and SARS by using technique of PCR and immunofluorescence by the Department of Medical Sciences. RESULTS: Eighty-nine patients who were enrolled in the present study. Peak of the incidence was 43% on April 2005 and there was no incidence of the influenza from December 2005 till April 2006. This may be due to the mass campaign of influenza vaccination or seasonal variation. CONCLUSION: Vaccination remains the primary strategy for the prevention of influenza, and the broadened recommendations should lead to protection of a larger portion of the population. PMID- 17427520 TI - Lipodystrophy and dyslipidemia in human immunodeficiency virus-infected Thai patients receiving antiretroviral therapy. AB - BACKGROUND: Lipodystrophy and dyslipidemia are increasing problems in HIV infected patients receiving antiretroviral therapy (ART). OBJECTIVE: Determine the prevalence and predicting factors for lipodystrophy and dyslipidemia in these patients. MATERIAL AND METHOD: A cross-sectional study was performed on HIV infected patients receiving ART in January 2006. Clinical assessment for lipodystrophy and blood tests for lipid profile were obtained. RESULTS: There were 56 patients with a mean age of 42.3 years and 71.4% were male. Lipodystrophy was detected in 66.1%. Baseline characteristics between patients with and without lipodystrophy were similar except for age, history of receiving d4T and undetectable HIV-RNA (p < 0. 05). There was a high prevalence of dyslipidemia (53.6%). By logistic regression, undetectable HIV-RNA (odds ratio (OR) 6.4, p = 0.016) and receiving d4T (OR 7.4, p = 0.022) are predicting factors for lipodystrophy. CONCLUSION: Lipodystrophy and dyslipidemia in HIV-infected Thai patients receiving ART are common. Clinical monitoring for these abnormalities during ART is highly recommended. PMID- 17427521 TI - Prevalence of metabolic syndrome and its relationship to weight in the Thai population. AB - BACKGROUND: Although the prevalence of the metabolic syndrome (MetS) has been well-documented in Western Caucasian populations, there are few studies in non Caucasian populations. The objectives of the present study were to estimate the prevalence of MetS and to find an optimal BMI cut-off value for defining obesity in the Thai population. MATERIAL AND METHOD: A sample of 307 men and 295 healthy women aged between 20 and 90 years (average age of 45 years) who came for a health check-up clinic in Khon Kaen, a northeast province of Thailand, were studied. The present study was conducted between 2003 and 2004. The modified ATP III criteria were used to estimate the age-and-sex specific prevalence of MetS, in which a BMI of > or = 27 kg/m(2) for men and 25 kg/m(2) for women were used in place of waist circumference. In the Thai population, these BMI cut-offs were equivalent to a percent body fat of 25% and 35% in men and women respectively. RESULTS: The overall prevalence of MetS was 15%, with no significant differences between men (15.3%) and women (14.6%). In men, the prevalence increased from 9.5% among the 20-39 age group to 24.7% among the 50+ age groups. In women, the respective prevalence was 7% and 29.5%. When BMI was removed from the classification ofMetS, the overall prevalence of "MetS-without-BMI" (still defined by the presence of at least 3 abnormalities) in both men and women was 7.8%. However the prevalence of MetS-without-BMI increased with higher BMI levels: among those with BMI < 25, the prevalence was 4.6% in men and 5.0% in women; among those with BMI > or = 25, the prevalence was 13% in men and 16% in women. CONCLUSION: The prevalence of MetS in this semi-rural Thai population was 15%, which is as common as in Caucasian populations. In the Thai population, obesity was a major component of MetS. PMID- 17427522 TI - Demonstration of nasogastric intubation using video compact disc as an adjunct to the teaching processes. AB - BACKGROUND: An in-house produced video compact disc (VCD) demonstrating nasogastric intubation was introduced to the medical curriculum at Buddhachinaraj Hospital, Clinical Teaching Centre, Phitsanulok in 2005. OBJECTIVE: The present study aimed to qualitatively evaluate the effect of the VCD on our medical students' procedural skills. MATERIAL AND METHOD: The 6th year medical students were randomly allocated into two groups; one group viewed the VCD (test, n = 30), and the other group did not (control, n = 31). After 2 months, the authors examined all students by asking them to perform the procedure using a manikin and a checklist. The examiners were blind to the allocations at the time of examination. Unpaired t-tests and Chi-square tests were used Data are expressed as mean and SD. RESULTS: Total checklist's score was 20. There was no diference in terms of age or gender between the two groups. Thirty of 61 medical students who had watched the VCD got an average score of 15.9 +/-1.86, while the control group's average score was 13.2+/-1.94. Statistical analyses showed that the difference was significant (p < 0.0001). CONCLUSION: The medical students were able to self-improve their procedural skills of nasogastric intubation after viewing the VCD. Application of this method to other basic procedures may be useful. PMID- 17427523 TI - Ventriculo-subgaleal shunt: step-by-step technical note. AB - Ventriculo-subgaleal (VSG) shunt placement is an option for temporary CSF diversion in neonate suffering from hydrocephalus. The authors describe step-by step technique of VSG shunt assembly and insertion by using previously-cut shunt tubes and connector. It is simple and inexpensive. The authors also present a short review of the literature of the VSG shunt. PMID- 17427524 TI - Pain reduction in patients with painful vertebral compression fractures undergoing percutaneous vertebroplasty. AB - BACKGROUND: Vertebral compression fracture (VCF) is the most common complication of osteoporosis. It results in significant mortality and morbidity. Percutaneous vertebroplasty (PVP) is a procedure that injects percutaneously bone cement into a collapsed vertebra. OBJECTIVE: To determine the results of PVP in pain reduction from osteoporosis VCF and its complications. MATERIAL AND METHOD: Thirty-five patients (34 women, 1 man, 48-98 years) with persistent back pain due to VCF underwent 66 percutaneous injection of polymethylmethacrylate (PMMA) into the vertebrae (27 thoracic levels, 39 lumbar levels) under fluoroscopic guidance between December 2003 and July 2005. Severity of back pain was assessed by using visual analog scale (VAS) before and after the operation. RESULTS: Thirty-two patients (91%) reported significant pain relief the mean VAS of 35 patients, before PVP and after an 8-week period, post-operatively, were 6.9 +/-1.8 and 2.0+/-1.8 (p = 0.001). There was only one minor complication. Two patients experienced intermittent sciatic shooting pain. This improved and disappeared within three months. CONCLUSION: P VP is a minimally invasive procedure providing safe, immediate, and sustained pain reduction in patients with refractory pain and disability caused by painful VCF PMID- 17427525 TI - Risk factors of deep vein thrombosis (DVT) after total knee arthroplasty (TKA) at Phramongkutklao Hospital. AB - BACKGROUND: There have been sporadic reports on the Asian risk factor of DVT after total knee arthroplasty. OBJECTIVE: To determine the risk factors of DVT MATERIAL AND METHOD: Retrospective review of one hundred patients who had undergone TKA and postoperative Contrast Venography in bilateral legs between 2002 and 2005 were performed to identify risk factor of DVT The patients were divided into two groups, positive and negative venography which the patients who had positive venography were indicating the development to DVT RESULTS: One hundred patients were evaluated associated to DVT Eighteen of these patients were men, and eighty-two were women. The median age at the time of the procedure was seventy-five years old (range: 62 - 79 years old). Sixty-one patients showed positive venography for D VT Five critical risk factors were identified to develop DVT: 1. Underlying cardiovascular disease 2. Underlying hematological disease 3. Underlying rheumatoid arthritis 4. Patients who took oral herbal medicine about one year before the operation 5. Patients who received revision TKA. CONCLUSION: The risk factors of DVT in the presented patients at Phramongkotklo Hospital were similar to other countries. The research study could identify statistically significant risk factors and stimulate surgeons undertaking TKA to be aware of the probability of the patient to develop DVT. PMID- 17427526 TI - Prospective study of hypotension and bradycardia during spinal anesthesia with bupivacaine: incidence and risk factors, part two. AB - OBJECTIVE: Hypotension and bradycardia during spinal anesthesia are common and may relate to severe adverse events such as cardiac arrest or death. The authors'first retrospective study identified three non-modfiiable factors including females, age more than 40 years, and type of operation. Level of blockage higher than T5 was the only one modifiable factor that could be confirmed That study was strongly limited by the nature of the retrospective anesthetic record reviewed study. MATERIAL AND METHOD: The authors prospectively studied the records of 2, 000 patients who received spinal anesthesia at Sriraj Hospital from 1 July 2004 to 31 December 2004. The collected parameters were patient demographic data (sex, age, body weight, height, ASA status), operative data (type of operation, emergency status, position and duration of operation), anesthetic data (type and dosage of local anesthetic agents used, intravenous fluid, vasoactive and sedative agents, highest sensory level of spinal blockage, usage and doses of spinal opioids). Recording forms of hypotension and bradycardia were completed by the responsible anesthetic team in each consecutive patient. RESULTS: Incidence of hypotension (20% or more decrease in systolic blood pressure) in the present study was 57.9%. The highest incidence was in cesarean section. Four non-modifiable risk factors could be identified which included females, age more than 50, body mass index more than 35 (OR = 2.1, 95% CI 1.01-4.29) and type of operation. Two modifiable risk factors included high dose of heavy bupivacaine (OR 1. 88, 95%CI 1. 3-2.74) and level of sensory blockage equal to or higher than T5 (OR 2.27, 95%CI 1.73-2.97). CONCLUSION: Usage of high dose of heavy bupivacaine and level of blockage higher than T5 were two modifiable risk factors associated with hypotension during spinal anesthesia. Avoidance of high block and lower dose of heavy bupivacaine can reduced the incidence and severity of hypotension after spinal anesthesia. PMID- 17427527 TI - The acromial morphology of Thais in relation to gender and age: study in scapular dried bone. AB - OBJECTIVE: The objective of the present study was to determine the acromial shape and examine if there is a correlation between the acromial morphology and genders, ages and sides. MATERIAL AND METHOD: The present examined 154 dried Thai scapulas (107 males and 47 females) with age range from 16 to 87years (mean = 49 + 17 years). The acromial morphology of each scapula was studied by the computerized image analysis of digitized photography through the supraspinatous outlet view, with the distance (M) measured from its anterior to posterior end, the height (H) of the resultant curve and the distance (N) from the anterior end to the point perpendicular to the height. The acromial types were defined as type I (flat), II (curved) and III (hooked) with the criteria that N is more than or equal to the 2/3, 1/3 and less than 1/3 of M, respectively. RESULTS: The incidences of types I, II and III were 3.2%, 93.5% and 3.2%, respectively. It was found in both sexes, female (93.6%) and male (93.5%) and in both sides, left (96%) and right (91.1%). With respect to the age range, type II were found in 100% of subjects of less than 30 years, 4.5% in those between 30-60 years, were 4.5% (1), 93.2% (1), 2.3% (111). Those more than 60 years old were 2.3 %(1), 90.7% (11) and 7.0% (III). The spur formation on the anterior end of the acromion was found in 14.9% of scapulas, curved type mostly; and it was associated with hooked type in only one scapula. CONCLUSION: There was no significant type difference between sex, side and age range (p > 0. 05). The spurs found are not related to acromial morphology and old age. PMID- 17427528 TI - Nonmydriatic digital retinal images for determining diabetic retinopathy. AB - OBJECTIVE: To evaluate the efficacy of nonmydriatic digital retinal images for determining diabetic retinopathy. MATERIAL AND METHOD: Single field 45-degree digital retinal images of 225 eyes from 142 diabetic patients were obtained with a nonmydriatic camera. The images were diagnosed and graded by a general ophthalmologist. These results were compared with clinical diagnosis obtained by retinal specialists, after examination by using biomicroscope with plus lens and indirect ophthalmoscope of the patients. International clinical diabetic retinopathy disease severity scale was used for grading diabetic retinopathy in all cases. RESULTS: Presence of diabetic retinopathy was detected in 70 eyes (31.1%). The sensitivity and specificity for determining diabetic retinopathy was 68.57% (95%CI 57.00-78.20) and 92.25% (95%CI 87.00-95.50), respectively. The positive predictive value and negative predictive value was 80.00% (95%CI 68.20 88.20) and 86.67% (95%CI 80.60-91.00). Overall accuracy was 84.89%. CONCLUSION: Single field 45-degree nonmydriatic digital retinal images were limited by fair sensitivity for determining diabetic retinopathy although overall accuracy from the present study was relatively high. Upcountry, this tool might facilitate increased access of diabetic patients for eye evaluation but cannot replace standard eye examination. PMID- 17427529 TI - Incidence of antibiotic-associated diarrhea in a pediatric ambulatory care setting. AB - BACKGROUND: Although antibiotic-associated diarrhea (AAD) is a common adverse event in children receiving oral antibiotics, few epidemiological studies have investigated this issue. OBJECTIVE: To determine the incidence of AAD in children who received oral antibiotics at the Pediatric Outpatient Department, Chiang Mai University Hospital. MATERIAL AND METHOD: Children who were prescribed oral antibiotics between September 2004 and June 2005 were randomly enrolled. Subjects with immunodeficiencies, acute/chronic diarrhea, and a history of having taken antibiotics within two weeks prior to this visit were excluded Patients' characteristics including age, gender principal diagnosis, and type of antibiotics were recorded. Parents were asked to observe stool frequency and consistency until one week after discontinuing antimicrobial agents and fill out an appropriate questionnaire. AAD was defined if there were at least three loose or liquid stools per day for two consecutive days. Risk factors including age, type, and dosage of the antibiotics used, were analyzed RESULTS: Two hundred and twenty-five children were eligible for data analysis. The mean age was 4.1 years (0.3-14.5 years). Pharyngotonsillitis was the most common diagnosis (53.8%), and amoxicillin and cloxacillin comprised the most common antibiotics prescribed in the present cohort. The incidence ofAAD was 6.2%. All episodes were presented while the patients were taking antibiotics with a mean (+/- SD) onset and duration of occurrence of 2.28 +/- 1.13 and 2.64 +/- 1.15 days, respectively. Premature discontinuation of antimicrobial agents was reported in nine patients (64.3%). There was a trend towards a higher incidence of AAD in the amoxicillin/clavulanate group (16.7%) compared to amoxicillin (6.9%) and erythromycin (11.1%) groups, although it was not statistically significant. In addition, the present study could not demonstrate an association between younger age or the high dosage of antibiotics used, and the development ofAAD. CONCLUSION: AAD was not uncommon in a pediatric ambulatory care setting. It tended to occur in younger children receiving amoxicillin/clavulanate. PMID- 17427530 TI - Cognitive-behavioral intervention for young tsunami victims. AB - OBJECTIVE: To evaluate the efficacy of the cognitive-behavioral intervention provided to children by volunteer professionals in a tsunami-hit area in Thailand. The intervention was aimed to prevent the severe development of post traumatic stress symptoms, and to help the children who might recover spontaneously to do so more quickly. MATERIAL AND METHOD: One hundred and sixty children in Ranong province voluntarily participated in 2-days group activities on the 57h day after the tsunami. The manualized intervention had been designed based on cognitive-behavioral model. Three domains of post traumatic stress reactions were targeted, intrusion, arousal, and avoidance. The Children's Impact of Events Scale (13) (CRIES-13) was completed by the participants before and 2 weeks after the intervention. RESULTS: The program was generally well understood by the children. No difference in CRIES scores was observed before and after the children entered the intervention program. However when the sample was categorized into two groups, the group that was more likely to develop PTSD showed a significant reduction in the scores, whereas a significant increase in the scores was evident in the other group. CONCLUSION: The findings supported the efficacy of cognitive-behavioral intervention in the children who were prone to develop PTSD. The program needed to be adapted to suit the religious, socio economic, and cultural background of the sample. The unchanged Avoidance scores in the present study were possibly explained by the general feeling that a tsunami might happen again and the parental involvement. The increase of post intervention scores in otherwise normal sample, though not exceeding the cut-off raised some concern, and possibly the need for screening before the intervention. PMID- 17427531 TI - The comparison of Montgomery and Asberg Depression Rating Scale (MADRS thai) to diagnostic and statistical manual of mental disorders (DSM) and to Hamilton Rating Scale for Depression (HRSD): validity and reliability. AB - OBJECTIVE: To assess the validity and reliability of the Montgomery and Asberg Depression Rating Scale (MADRS Thai) by comparing DSM-IV TR criteria and Hamilton Rating Scale for Depression (HRSD). MATERIAL AND METHOD: Descriptive study. Subjects were recruited from the psychiatric out-patient clinic Siriraj Hospital. Subjects were assessed by MADRS Thai version and clinical interview. VALIDITY: Opinion from psychiatric and social science experts, evaluating 13 study cases and 2 7 control cases, compared to the Global Assessment Scale (GAS) and to the changed scale of HRSD. RELIABILITY: Inter-rater and Intra-rater reliability and Internal consistency. RESULTS: Content validity is above 0.5 except the item oflnner tension, Lassitude and Inablilty to feel. Criterion validity when compared to DSM-IV TR or HRSD, the sensitivity, specificity, PPV and NPV were all 100%. Pearson correlation coefficients sensitive to change were -0. 49 (p -value = 0. 11) and 0. 679 (p-value = 0. 025) when compared to 1) DSM-IV TR and 2) HRSD, respectively. ICC oflnter-rater and Intra-rater reliability were both 0.99 when compared to either 1) DSM-IV TR or 2) HRSD. Cronbach 's alpha of Internal consistency was 0. 96 and 0. 92 when compared to 1) DSM-IV TR and 2) HRSD, respectively. CONCLUSION: The result suggested that the Thai version of the Montgomery and Asberg Depression Rating Scale had a very good validity and reliability. Therefore, it can be used as a diagnostic test in Thai depressed patients. PMID- 17427532 TI - Reliability of the evaluation for left ventricular ejection fraction by ECG-gated multi-detector CT (MDCT): comparison with biplane cine left ventriculography. AB - OBJECTIVE: To evaluate the reliability of measurement for left ventricular ejection fraction (LVEF) by ECG-gated multi-detector CT (MDCT) comparing with biplane cine left ventriculography that is current gold standard. MATERIAL AND METHOD: The authors reviewed the data from 15 patients who were referred for coronary CT angiography for clinical indications and underwent cardiac catheterization within 14 days. Coronary CTA studies were performed on MDCT Somatom Sensation 16, Siemens, Germany, Slice thickness 1 mm, Slice collimation 0. 75 mm, and Pitch 0. 3. L VEF were measured with MDCTby Simpson s method and compared with values measured by biplane area length method from cardiac catheterization. The L VEF from both techniques were compared using intraclass correlation power analysis (SPSS analysis software). RESULTS: The study population consisted with six men and nine women with a mean age of 54+/-10 years. The LVEF measured from MDCT and cine ventriculography were 54.7 +/-10% and 56. 3+/-10%, respectively. LVEF measured with MDCT by interpreter I and interpreter 2 was significantly correlated with L VEF measured with biplane cine ventriculography (ICC= 0.99 and 0.98, respectively). The interobserver reliability was excellent with ICC = 0.9. CONCLUSION: LVEF measurement with MDCT during coronary CT angiography can be performed easily, very accurately, and compare well with measures taken from biplane cine left ventriculography. PMID- 17427533 TI - Sensitivity of mammography and ultrasonography on detecting abnormal findings of ductal carcinoma in situ. AB - OBJECTIVE: To analyze the mammographic and ultrasonographic findings of ductal carcinoma in situ (DCIS) and determine the sensitivity in Thai women. MATERIAL AND METHOD: Mammograms and bilateral whole-breast ultrasonograms of 37 proven cases of DCIS were reviewed. The former was assessed for microcalcifications and soft tissue densities while the latter was evaluated for masses and thickened ducts. Ultrasonography was used to spot the areas to visualize soft tissue densities in mammogram. RESULTS: Mammography detected 22 cases of DCIS having pure microcalcifications, eight cases with mixed microcalcifications and soft tissue densities, six cases with pure abnormal soft tissue densities and one case showing negative finding. The ultrasonography detected 13 cases showing masses, seven cases as showing thickened ducts and 17 cases as negative findings. CONCLUSION: Microcalcifications are characteristic findings in mammogram accounting for 81% of DCIS in the present study. Ultrasonography shows abnormalities including mass and thickened duct lesions in 54% of DCIS. The combined modalities can give the detection of abnormalities in 97% of DCIS. PMID- 17427534 TI - Leptospirosis in Takeo Province, Kingdom of Cambodia, 2003. AB - BACKGROUND: In Cambodia, epidemiology and disease burden of leptospirosis were not addressed as they do not have an existing surveillance system and have limitations on their laboratory diagnosis. OBJECTIVE: Define the existence of leptospirosis and determine the antibodies to serovars of leptospires in Cambodia. MATERIAL AND METHOD: One hundred and twenty-one suspected cases of leptospirosis were enrolled in this cross-sectional study, between September 8 and November 30, 2003 from Takeo Provincial Hospital in Doun Keo District, Cambodia. RESULTS: Common clinical manifestations were fever (96%), headache (92%), and myalgia (87%). Common risk behaviors were throwing garbage on the ground (84%), pulling out sprouts (77%), fertilizing (49%), and plowing (47%). Microscopic agglutination test result confirmed four cases and polymerase chain reaction test result confirmed seven cases. Two cases each showed antibodies to serovars Javanica and Australis. An estimated annual incidence of leptospirosis in Takeo province was 7.65 per 100,000 populations. Further studies to define epidemiology and burden of disease are needed. CONCLUSION: Increasing awareness and knowledge on leptospirosis among people are necessary to decrease the impact of leptospirosis in Cambodia. PMID- 17427535 TI - Adverse reactions of 300 MG diethylcarbamazine, and in a combination of 400 MG albendazole, for a mass annual single dose treatment, in migrant workers in Phang Nga province. AB - BACKGROUND: Foreign migrant workers with work permits in Thailand are given once a year 300 mg diethyl-carbamazine (DEC) for bancroftian filariasis, and 400 mg albendazole (ABZ) for helminthiasis. Treatment effectiveness, tolerability, and safety of two treatment arms, DEC + ABZ and DEC alone, had never been fully documented. OBJECTIVE: Evaluate the tolerability of the two treatment arms and analyze the effects of adverse reaction, prevalence, and intensity of both common and uncommon adverse drug reactions (ADR) in relation to the reaction time (2 hours = acute, > 2 to 24 hours = subacute, and > 24 to 72 hours = latent). MATERIAL AND METHOD: A hospital-based clinical study of on-hour-2 treatment with both treatment arms in 280 Myanmar male migrant volunteers (DEC + ABZ = 150, DEC = 130) was conducted in Phang Nga province, southern Thailand Of these, ADR evaluation at three reaction times was performed using antigenemic (WbAg+) and non-antigenemic (WbAg-) volunteer groups (DEC + ABZ/WbAg+ = 14, DEC/WbAg+ = 12, DEC + ABZ/ WbAg- = 8, andDEC/WbAg- = 16). RESULTS: Both drug groups had similarly overall ADR prevalence [5.2%for DEC + ABZ and 5. 1% for DEC (p > 0. 05)], as well as mean ADRacute scores (p > 0. 05) on hour 2 post-treatment. The four groups had maximum overall prevalence (10% to 40%for ADRsubacute). It was more likely to show no relationship between treatment arms and WbAg (neither WbAg+ nor WbAg-) with adverse reaction intensity for ADRacute, ADRsubacute, Or ADRlatent Three major specific ADR were fatigue, dizziness, and headache. CONCLUSION: Adverse reaction prevalence and intensity were independent for WbAg and treatment arm. The DEC + ABZ have no greater effects on ADR development as the DEC does. The common ADR after treatment are not required for symptomatic treatment. The study confirms DEC + ABZ regime can be safe and not toxic for use in mass treatment of those migrants in Thailand and, its value, in a mass annual single dose treatment, is beneficial for the Global Alliance to Eliminate of Lymphatic Filariasis (GAELF). PMID- 17427536 TI - Bioequivalence study of 30 mg pioglitazone tablets in Thai healthy volunteers. AB - OBJECTIVE: To compare the bioequivalent parameters of 30 mg pioglitazone tablets manufactured locally (Glista) and originally (Actos). MATERIAL AND METHOD: A randomized, single dose, two-treatment, two-period, two-sequence crossover study was conducted Twenty-four healthy volunteers were recruited at Siriraj Clinical Research Unit. Each subject received a 30 mg pioglitazone tablet of both formulations with at least a week washout period Blood samples were collected over 48 h after the oral administration. The plasma fractions were analyzed for pioglitazone using a liquid chomatography-mass spectrometry (LC-MS/MS). RESULTS: Twenty-four volunteers enrolled in the present study. Pharmacokinetic parameters were determined using the non-compartment model. The 90 percent confidence intervals of the mean ratios (test/reference) of Cmax (86.2687-113.7313%), A UC0- >t(85. 7139-114.2861%) and AUC0-->infinity (81.7820-118.2180%) fell within the acceptable range (80-125%) for bioequivalent eligibility. Both preparations were well tolerated and had afew non-serious adverse events. CONCLUSION: The 2-tablet preparations of pioglitazone were bioequivalent in Thai healthy volunteers. PMID- 17427537 TI - Advancing age and renal impairment as important predisposing factors of gatifloxacin-induced hyperglycemia in non-diabetes patients. AB - There have been case reports about adverse effects to glucose homeostasis related to gatifloxacin use. The authors report an elderly, non-diabetic patient who developed severe hyperglycemia after receiving oral gatifloxacin 400mg/d. He was a 73-year-old male, patient with a history of hypertension, cured vesical pheochromocytoma, idiopathic dilated cardiomyopathy, chronic renal insufficiency (baseline serum creatinine of 1.7 mg/dL), and gouty arthritis admitted to the hospital with a diagnosis of acute bronchitis. Seven days after initiating gatifloxacin, his symptoms were improved. Subsequently he developed polyuria, polydipsia, and fatigue with an increase in serum creatinine to 2.8 mg/dL, and random plasma glucose levels elevated to 903 mg/dL. Gatifloxacin was stopped. Intravenous regular insulin infusion was administered. Euglycemia was achieved within 8 hours after fluid rehydration and only low dose insulin was required He maintained normal glucose levels without any antidiabetic drugs afterward. Old age and renal impairment were considered significant contributing factors for this hyperglycemic adverse event from gatifloxacin. PMID- 17427538 TI - Case report combined hepatocellular and cholangiocarcinoma with sarcomatous transformation. AB - Combined hepatocellular and cholangiocarcinoma with sarcomatous transformation was first recognized in Ramathibodi Hospital in 2005. This variant of carcinoma has been increasingly reported particularly from Asian countries. Dedifferentiation of the epithelial component to various sarcomatous components is likely the underlying mechanism. The causative factors of hepatocarcinogenesis in Thailand include chronic viral hepatitis B or C, exposures to aflatoxin B1 and nitrosamine(s) and occasionally some certain nodular hepatocellular lesions due to arterial hyperperfusion. It is suggested that the recent change of the Thai peoples' life style to an increased consumption of fast foods containing food preservatives especially nitrate or nitrite, the nitrosamine precursor may allow heavy exposure(s) to the chemical carcinogen(s) i.e. nitrosamine(s) leading to sarcomatous transformation of the carcinoma. PMID- 17427539 TI - Total anal sphincter saving technique for fistula-in-ano; the ligation of intersphincteric fistula tract. AB - OBJECTIVE: To describe a new technique for fistula-in-ano surgery aimed at total sphincter preservation, and evaluate the preliminary results concerning non healing and intact anal function. MATERIAL AND METHOD: A prospective observational study in eighteen fistula-in-ano patients treated by ligation of intersphincteric fistula tract (LIFT) technique, from January to June 2006. RESULTS: Fistula-in-ano in seventeen patients healed primarily (94.4%). There was one non-healing case (5.6%). The mean healing time was four weeks. None had disturbances in clinical anal continence. CONCLUSION: The early outcome of the LIFT technique is quite impressive. Results warrant a larger study with long-term evaluation. This technique has the potential to become a viable option for fistula-in-ano surgery. PMID- 17427540 TI - Epilepsy and the law--a view from Thailand. AB - BACKGROUND: Patients with epilepsy often suffer social injustice in many societies. OBJECTIVE: This is a review of current laws dealing with epilepsy and the consequences of this complex disease. MATERIAL AND METHOD: The author looked at reported unjust legal and social acts resulting from epileptic seizures and their results. RESULTS: The author found that accusation of criminal acts, accidents while driving, and job loss are relatively common experiences for epileptics. Such events reflect lack of public understanding. There is inadequate legislation to protect epileptics. Most disturbing areas are ictal behavior misinterpreted as crime, driving while experiencing a seizure, and social discrimination. There is no law dealing with epilepsy in Thailand while considerable progress in this field has been made in Western countries. CONCLUSION: Epilepsy patients are inadequately protected by Thai law. There is an urgent need for education and campaigns for their rights focused on the public, the legal profession, and the health care professionals. PMID- 17427541 TI - Update on status epilepticus. AB - Status epilepticus is a medical emergency and is secondary to a range of insults to the central nervous system. The authors reviewed the current management of this disorder in light of the latest developments from recent trials and guidelines. Important principles in management include early recognition of status epilepticus, identification of the underlying cause and prompt treatment to terminate seizures and reduce complications. The role of electroencephalographic monitoring and different treatment regimens are examined. PMID- 17427542 TI - A fast approaching crisis. PMID- 17427543 TI - A traditional model of pregnancy care shows promise in reaching disadvantaged women. PMID- 17427544 TI - Specialty pharmacists boost the care and compliance of HIV-infected patients. AB - Pharmacists have a key role to play in managing patients infected with HIV That is the conclusion of a new study showing that clinical pharmacists may well be an underutilized resource in managing patients Swho are beginning treatment with highly active antiretroviral therapy. And investigators report that their value is particularly noticeable among disadvantaged populations. PMID- 17427545 TI - Care managers have more resources at their disposal in new Medicaid effort in Florida. AB - New multidisciplinary care model gives Pfizer another crack at Florida's Medicaid population. Two years after an earlier DM program was discontinued, Florida is once again turning to Pfizer Health Solutions (PHS) to handle its biggest DM program to date. This time, PHS is bringing more resources to the table and taking on an additional three disease states. PMID- 17427546 TI - The Joint Commission calls for comprehensive action on poor health literacy. AB - A new call to action on poor health literacy. Maintaining that the problem is costing the country dearly in dollars as well as quality of life, The Joint Commission has put out a new policy white paper calling on all key players to take steps to deal with the problem of poor health literacy. Although changes at the top are sorely needed, policy makers maintain that there is much that individual providers can do to alleviate the problem. PMID- 17427547 TI - [In vitro susceptibility of respiratory isolates of Streptococcus pneumoniae and Streptococcus pyogenes to telithromycin and 11 other antimicrobial agents: Turkish results of e-BASKETT-II surveillance study]. AB - In respiratory tract infections, therapy is often empirical and there is a need for local data on the rate of resistance to available antimicrobials. In this multicentre study which is a part of the international e-BASKETT-II surveillance study, respiratory isolates of Streptococcus pneumoniae (n=260) and Streptococcus pyogenes (n=312) collected between September 2002 and June 2003 from 18 hospitals in Turkey were tested against penicillin G, amoxicillin, cefuroxime, ceftriaxone, erythromycin, clarithromycin, azithromycin, clindamycin, telithromycin, tetracycline, levofloxacin and vancomycin. Antibiotic susceptibilities were determined with disk diffusion method and confirmed with broth dilution method following the CLSI guidelines. Isolates which were resistant to erythromycin were genotyped by polymerase chain reaction. In S. pneumoniae 11.5% of the isolates were highly and 22.7% were intermediately resistant to penicillin. Rate of resistance to erythromycin, clarithromycin, azithromycin was 17.3%, and 21.5% of the isolates were resistant to tetracycline. Resistance to levofloxacin and vancomycin was not observed and only one isolate was found intermediately resistant (MIC=2 microg/mL) to telithromycin. Genotypes in erythromycin-resistant isolates were ermB (77.8%), mefA (17.8%) and ermB+mefA (2.2%). S. pyogenes isolates were uniformly susceptible to beta-lactams and vancomycin, and only one isolate was intermediately resistant to levofloxacin. Macrolide resistance was observed in 1.3% of the isolates and three out of these harboured the mefA gene. One isolate with an MIC of 4 microg/mL for telithromycin had ermB gene. Telithromycin has demonstrated a good in vitro activity against macrolide resistant respiratory tract isolates. As a result, e-BASKETT-II surveillance study has been one of the most extensive in vitro studies comparing telithromycin to available antimicrobial agents for respiratory tract infections in Turkey. PMID- 17427548 TI - [Rapid purification of Bordetella pertussis fimbria 2, 3 proteins and their immunogenicities]. AB - Bordetella pertussis fimbria antigens play an important role in the adhesion to the cell surfaces, and induce the formation of protective antibodies in the host. The aim of this study was to purify the fimbria proteins by a rapid and easy method which involved Superdex 75 gel filtration chromatography, and to investigate their immunogenicity. The fimbria proteins purified from the different strains showed single protein bands in 12.5% SDS-PAGE (Sodium dodecyl sulphate-polyacrylamide gel electrophoresis) method. Depending on the strain used, the proteins were observed as Fim2 and Fim3 subunits or only Fim 2 subunit on 16% SDS-PAGE. The purified subunits were confirmed by Western and dot blotting analysis with monoclonal antibodies BP F2 (anti-Fim2) and BPC10 (anti-Fim 3). The immunogenicity studies performed in the mice revealed that purified fimbria proteins have led to higher agglutinin response than the vaccine with whole cell pertussis antigens. It was determined that the extent of agglutinin responses were different for different strains from which the fimbria proteins were purified. In addition, each B. pertussis strain showed a different agglutination reaction with different anti-Fim sera. The results, overall, pointed out the significance of fimbria structure of the B. pertussis strain used for vaccine manufacture and control. PMID- 17427549 TI - [Biotypes and antibiotic resistance patterns of Gardnerella vaginalis strains isolated from healthy women and women with bacterial vaginosis]. AB - As Gardnerella vaginalis is accepted as a member of normal vaginal flora, it is one of the dominant species which has been related to bacterial vaginosis (BV). The aim of this study was to determine the isolation rate, biotypes and antibiotic resistance patterns of G.vaginalis from the vaginal swab samples of 408 women who were admitted to the outpatient clinics of Family Planning Center. Hippurate hydrolysis, lipase and beta-galactosidase tests were performed for biotyping the isolates, and agar dilution (for metronidazole) and disk diffusion (for clindamycin) tests were used for the detection of antibiotic resistance patterns. As a result, by Nugent's BV scoring protocol, 122 (29.9%), 20 (29.4%), 137 (33.6%), and 18 (4.4%) of the women were diagnosed as BV, intermediate form, normal vaginal flora (NVF) and mycotic vaginosis, respectively. The overall isolation rate of G.vaginalis was found as 23% (94/408). Of them, 56.4% (53/94) and 8.5% (8/94) were isolated from samples of BV cases and subjects with NVF, respectively, and the difference was statistically significant (p<0.05). The biotyping results showed that the most frequently detected types were biotype 1 (44%), 5 (20%) and 4 (18%). There was no statistically significant difference between the biotype distribution of BV patients and the subjects who have NVF (p=0.687). The results of antibiotic susceptibility tests indicated that 70% and 53% of the isolates were resistant to metronidazole and clindamycin, respectively. It was of interest that MIC values for metronidazole was > or =128 microg/ml in 57% of resistant strains. The data of this study has emphasized that the metronidazole resistance is very high in our population, and the large scale studies are needed to clarify the relationship between BV and G.vaginalis biotypes, which can be found in the normal vaginal flora. PMID- 17427550 TI - [Investigation of the presence of Clostridium difficile in antibiotic associated diarrhea patients by culture and toxin detection methods]. AB - Clostridium difficile-associated disease can be observed especially in hospitalized patients who use broad-spectrum antibiotics. The aim of this study was to investigate the presence of C. difficile as the causative agent of diarrhea in outpatients and inpatients. During January-December 2005, 45 outpatients and 46 inpatients (of them 11 were intensive care unit patients) who had developed diarrhea due to antibiotic use, were included to the study. In addition 7 intensive care unit personnel and 20 food handlers were also included to the study in order to detect their carrier states. The age range of patients was 16-80 years, and of them 45 (49.5%) were male, while the age range of the personnel was 25-55 years, and of them 21 (78%) were male. Stool samples collected from the study groups were cultivated in C. difficile agar media (C. difficile Agar Base, Oxoid) as well as on routine bacteriologic media, and C. difficile growth was confirmed by latex agglutination test with the use of specific antisera. The presence of C. difficile toxin A was investigated by latex method (Oxoid, UK), and toxin A and B was searched by enzyme-linked immunoassay (ELISA; Seramun GmbH, Serazym C. difficile Toxin A+B), in the stool samples. While C. difficile was isolated from 13 (14.3%) of the 91 samples, no positive result was detected in the personnel. There was no statistically significant difference between outpatient and inpatient groups by means of C. difficile culture positivity (15.5% and 17.1%, respectively) (p>0.05). All of the culture positive samples were also found positive by ELISA Toxin A+B method (100%), but only 4 of them (30.7%) yielded positive result by Toxin A latex test. It was detected that 84.6% (11/13) of the patients had used ampicillin/sulbactam, 7.7% (1/13) used cotrimoxazole-SXT, and 7.7% (1/13) used macrolide antibiotics. The use of ampicillin/sulbactam was found statistically significant in development of diarrhea (p<0.05). Our data indicated that ELISA Toxin A+B is a reliable method with 100% specificity and sensitivity in the rapid diagnosis of C. difficile until the culture results were obtained, however, although specificity of Toxin A latex test is 100%, its use alone as a primary rapid diagnostic test was not recommended because of its low (30.7%) sensitivity. PMID- 17427551 TI - [Nosocomial urinary tract infections in the intensive care unit patients]. AB - The aim of this prospective study was to determine the frequency of nosocomial urinary tract infections (UTI) and risk factors in intensive care unit patients. A total of 110 patients were included in the study, and UTI developed in 39.1% of them. The mean age of patients with UTI (53.6+/-20.0 yrs) was found higher than the patients without UTI (39.7+/-22.2 yrs), (p<0.01), whereas there was no gender associated difference between these two groups (p>0.01). No patient had developed symptoms suggesting UTI. All of the UTI has developed in patients with urinary catheter. Infections occurred in 72.2% of the patients with catheter lasting for seven days or more, and 24.3% of those lasting less than seven days (p<0.001). The UTI development rate was found as 31.3% in the patients who had been using antibiotics, while this ratio was 50% in those who had not used antibiotics (p<0.05). Pyuria was detected in 86.1% and 17.9% of the patients with and without UTI, respectively (p<0.001). Detection of bacteria in urine by the examination of Gram stained preparations, and the tests of peripheral blood leucocyte counting and CRP levels were not found sensitive and specific for nosocomial UTI. The prevalence of UTI increased gradually by the duration of hospitalization. The most frequently isolated microorganisms were found as Candida (27.2%), Escherichia coil (27.2%) and staphylococci (12.9%). Since nosocomial UTI which are important causes of morbidity in intensive care unit patients, are difficult to diagnose and treat, more efforts should be used for preventive measures. PMID- 17427552 TI - [Investigation of Chlamydia trachomatis positivity in women with and without gynecologic complaints by cytologic and direct immunofluorescence methods]. AB - The asymptomatic nature of the majority of Chlamydia trachomatis infections leads to persistent infections and serious complications as well as continuous transmission of bacteria in the populations. The aim of this study was to investigate the presence of C. trachomatis in non-pregnant women with and without gynecologic signs and symptoms, and to detect the rate of asymptomatic carriage. Cervical specimens collected from 200 nonpregnant women (age range: 20-81 yrs; mean age: 40.2+/-10.4 yrs) who were admitted to Gynecology and Obstetrics Clinics of Hacettepe University Hospital were included to the study. Of them 68 had clinical complaints such as vaginal discharge, itching/irritation, inflammation and inguinal pain, while 132 had not any clinical complaints. All the samples were examined by direct immunofluorescence (DFA) method (Fluorotect Chlamydia, Omega Diagnostics, UK) with fluorescein isothiocyanate labeled monoclonal antibodies against C. trachomatis serotype specific major outer membrane proteins, and the samples were simultaneously screened cytologically by Papanicolaou staining method. As a result, C. trachomatis antigen positivity was found in 49 (24.5%) of the samples by DFA method, and chlamydial inclusion bodies were detected in 19 (9.5%) of women by cytologic method. Twelve (24.5%) of the 49 DFA positive samples, and 7 (4.6%) of the 151 DFA negative samples yielded positive results cytologically. The observed proportion of overall agreement (P) was estimated as 78% between the results of methods. C. trachomatis antigen positivity was detected in 16.2% (11/68) and 28.8% (38/132) of women with and without clinical symptoms, respectively. There was no statistically significant difference between C. trachomatis positivity and neither the presence of clinical signs and symptoms nor the characteristics of the signs and symptoms (p>0.05). In conclusion, the high asymptomatic carriage rate detected in our study population indicated that, for the prevention of bacterial transmission in the populations, the women who were admitted to gynecology and obstetrics clinics should be screened for C. trachomatis positivity even if they had no clinical complaints. The use of DFA method together with the widely used, practical and economical cytologic examination method, would increase the sensitivity and specificity of C. trachomatis diagnosis. PMID- 17427553 TI - [Differential diagnosis of bacterial and viral meningitis in childhood acute meningitis: a statistical model]. AB - Acute bacterial meningitis (BM) which is a pediatric emergency with high mortality and morbidity, must be diagnosed and treated promptly. There is no unique method to prove or rule out the diagnosis of BM in a patient with cerebrospinal fluid (CSF) findings consistent with BM but negative Gram stain and culture results. For this purpose the combination of CSF parameters are used for diagnosis. The aims of this study were to compare retrospectively the mean leukocyte count, serum C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), CSF leukocyte and neutrophil, CSF protein and glucose values in 40 bacterial and 29 viral meningitis (VM patients, ages between 1 month and 14 years, and to develop a statistical model for the differentiation of BM and VM cases. Logistic regression analysis was used to investigate the relationship between BM and age, CPR, ESR, leukocyte count, CSF leukocyte, neutrophil, protein and glucose values. Based on CSF protein and neutrophil ratio which were found as independent variables, the regression model could predict the patients having BM with 95% and viral meningitis with 93.2% accuracy. PMID- 17427554 TI - [Investigation of transplacental transmission of TT virus in mother--newborn pairs]. AB - TT virus (TTV) is widespread throughout the world and can be detected in 50-95% of healthy individuals. However, in chronic hepatitis C virus (HCV) infected patients coinfected with TTV, histological activity indices were higher than patients with HCV infection alone. There are studies which indicate that TTV can cause aplastic anemia and thrombocytopenia. While TTV is known to be transmitted through blood transfusions and by fecal-oral route, published information on transplasental transmission is controversial. The aims of this study were to detect the frequency of TTV infection among healthy pregnant women and to search whether TTV is transmitted transplasentally during pregnancy. For this purpose, plasma samples collected from 54 women and their newborns were investigated by polymerase chain reaction (PCR), and viral loads were determined for infected newborns, and their respective mothers. Also, ten mothers whose newborns were tested negative for TTV-DNA were enrolled in viral load tests for comparison. TTV DNA was detected in 49 (90.7%) of 54 women. Of 49 newborns whose mothers were infected with TTV, only 4 (8.2%) were found positive for TTV-DNA. There was no statistically significant difference in TTV-DNA loads between mothers who transmitted the virus and those who did not (Mann-Whitney U analysis Z=-0.071, P=0.944). As a result, transplasental transmission of TTV is possible but occurs in low frequency and independent of viral load. PMID- 17427555 TI - [Evaluation of 35 adult measles cases detected in a measles outbreak]. AB - The aim of this study was to evaluate the clinical, epidemiological and demographical features of adult measles cases admitted to our hospital, during the measles epidemic which emerged in Europe and in our country in the year 2001. A total of 35 adult measles cases (10 male, 25 female) ages between 16-36 (mean age: 23.1 +/- 4.6) years were detected between January and June 2001. The diagnosis was based on the clinical findings, however only 11 of the cases could be serologically confirmed with the presence of measles IgM antibody positivities. Fever (91.4%), cough (94.3%) and conjunctivitis (77.1%) were the most common symptoms. All of the patients had maculopapular rash, and the presence of Koplik spots, lymphadenopathy and hepatomegaly were observed in 77.1%, 57.1%, and 5.7% of patients, respectively. Leukopenia and elevated liver enzymes were detected in 28.5% and 37.2% of the patients, respectively. Pneumonia was the most common complication which was seen in 20% of the patients. Of the cases 2.9% exhibited otitis media and 17.1% diarrhea. Spontaneous abortus occurred in one of the three pregnant women. In conclusion, measles is still an important public health problem in our country, and since it may lead to severe complications and economic and labor loss, an effective elimination programme should be obtained by the use of vaccine in two doses and effective strategies for the immunization of the target populations. PMID- 17427556 TI - [Can HBsAg levels guide to differentiate inactive HBsAg carriers from HBeAg negative chronic B hepatitis?]. AB - It is valuable to differentiate the inactive HBsAg carrier state from HBeAg negative chronic B hepatitis (CBH) which develops due to precore or core promoter region mutations in hepatitis B virus (HBV). The aim of this study was to investigate the role of HBsAg S/N (sample rate/index calibrator mean rate) levels in the differentiation of inactive HBsAg carriers from HBeAg negative CBH cases. A total of 134 HBsAg positive patients followed-up in Kocaeli University Medical Faculty hospital between June 2004-September 2005, were included to the study. The patients were classified into four groups according to their serological patterns (Group 1: HBeAg and HBV-DNA negative 34 cases with normal ALT levels; Group 2: HBeAg negative, anti-HBe positive, HBV-DNA >10(5) copies/ml, 36 cases with increased ALT levels; Group 3: HBeAg negative, anti-HBe positive, HBV-DNA 102-10(5) copies/ml, 32 cases with normal ALT levels; Group 4: HBeAg positive, HBV-DNA >10(5) copies/ml, 32 cases with increased ALT levels). The age and gender distributions of the groups were similar. HBV markers have been detected by microparticle enzyme immunoassay (AxSYM System, v3.0, Abbott Laboratories, USA), and viral load were investigated by real-time polymerase chain reaction (iCycler IQ, v3.0a, Bio Rad Laboratories, USA). As a result, the mean HBsAg S/N level in group 2 who were HBeAg negative with a viral load of >10(5) copies/ml, was found significantly higher than group 1 who were inactive HBsAg carriers (285.9+/-78.8 and 214.4+/-108.6, respectively; p<0.05). In contrast there was no statistically significant difference between group 1 (HBV-DNA negative) and group 3 (HBV-DNA <10(5) copies/mL) by means of mean HBsAg S/N levels (214.4+/-108.6 and 216.3+/ 57.2, respectively; p>0.05). Although HBsAg levels seem to guide the differentiation of inactive HBsAg carriers from HBeAg negative CBH cases with high viral loads (>10(5) copies/ml), advanced studies are needed to clarify this relationship with the use of quantitative HBsAg measurements (IU/ml) in large patient groups and by performing mutation analysis. PMID- 17427557 TI - [Retrospective evaluation of patients who were diagnosed as infectious mononucleosis between 1984-2005]. AB - In this study, the clinical and laboratory features of 26 infectious mononucleosis (IMN) cases who have been diagnosed between the years of 1984-2005 were evaluated retrospectively. The mean age of the patients was 26+/-11 years, the rate of being hospitalized was 65%, and mean hospitalization period was 9.2+/ 6 days. Fever (81%), weakness (50%), sore throat (50%), headache (50%) and swollen neck (35%) were the most common symptoms, while in the physical examination cervical lymphadenopathy (81%), splenomegaly (69%), hyperemic pharynx (65%), hepatomegaly (54%) and tonsillitis (50%) were observed. Laboratory results yielded leukocytosis in 21%, leucopenia in 12%, anemia in 44%, thrombocytopenia in 5% and elevated transaminase levels in 84% of the patients. Of the patients 15 (57.7%) had the history of using antibiotics before the diagnosis. Serological diagnosis was performed by Paul-Bunnel test and/or IgM positivity against Epstein Barr virus (EBV) viral capsid antigen (VCA). Tonsillo-pharyngitis secondary to edema and respiratory distress due to lymphadenopathy pressure were detected in four patients, whereas pancytopenia was established only in one patient, as complications. This study emphasized that, although IMN is a self-limited infection, the diagnostic difficulties may arise when the clinical course is atypical, and rarely seen life-threatening complications may also develop during IMN course. PMID- 17427558 TI - [Development of a database for tracking HIV positive/AIDS patients]. AB - The collection of reliable data is the first step to assess the status of HIV/AIDS in a community. HIV recording systems are necessary for organizing and analyzing the patients' data. The aim of the study was to develop a database to be used to track HIV positive/AIDS patients. The database includes general demographic fields as well as specific fields such as health history, laboratory and other clinical history, current and past drug regimens (both antiretroviral and non-antiretroviral drugs). It is also possible to organize and maintain a patient database according to specific diseases, laboratory tests and/or medication treatments. PMID- 17427559 TI - [Short communication: use of cefoxitin disc diffusion method for the detection of methicillin resistance in Staphylococcus aureus clinical isolates]. AB - The aim of this study was to compare the cefoxitin disc diffusion method with oxacillin broth microdilution and agar screening methods for the detection of methicillin-resistant Staphylococcus aureus (MRSA). The presence of PBP2a in 94 S. aureus clinical isolates were screened by latex agglutination MRSA-Screen test (Oxoid Ltd, UK), and 64 (68%) of the strains were found positive. Cefoxitin disc diffusion method and oxacillin broth microdilution and agar screening methods were performed in the guidance of Clinical and Laboratory Standards Institute (CLSI) proposals. All of the 64 PBP2a positive isolates were found resistant, and 30 PBP2a negative isolates were detected as susceptible to oxacillin by both of the broth microdilution and agar screening methods. In cefoxitin disc diffusion method, zone diameters of 62 isolates were measured as 0-14 mm, while two isolates gave 15-17 mm and 30 isolates gave 220 mm zone diameters. According to the CLSI proposals, 64 isolates were accepted as oxacillin-resistant in view of their cefoxitin zone diameters. As a result, the sensitivity and specificity of cefoxitin disc diffusion method was detected as 100%, when comparing with both gold standard (PBP2a presence) method and oxacillin broth microdilution and agar screening methods, indicating that it is a reliable test to investigate oxacillin resistance of clinical S. aureus isolates in routine laboratories where molecular tests could not be performed due to economical reasons. PMID- 17427560 TI - [Short communication: comparison of susceptibilities of Escherichia coli urinary tract isolates against fosfomycin tromethamine and different antibiotics]. AB - The aim of this study was to investigate the susceptibilities of Escherichia coli strains isolated from urine samples, against fosfomycin tromethamine and different antibiotics in the period of October-December 2004 in a local hospital in Ankara, Turkey. A total of 120 E. coli strains isolated from urine cultures of subjects who were admitted to outpatient clinics were included to the study. The identification and antimicrobial susceptibility tests (against amikacin, amoxicillin/clavulanate, ampicillin, cefepime, cefoxitin, cefotaxime, cefuroxime, cefalotin, ciprofloxacin, gentamicin, levofloxacin, meropenem, nitrofurantoin, piperacillin, piperacillin/tazobactam and trimethoprim/sulphametoxazole) were performed by a commercial automatized system (Phoenix, Becton Dickinson, USA). Fosfomycin tromethamine susceptibility was studied by Kirby Bauer disk diffusion method according to the CLSI criteria. Only one strain (0.8%) was found resistant to fosfomycin tromethamine, while no resistance was determined against amikacin and meropenem. Most of the isolates were found susceptible to nitrofurantoin (90%), cefoxitin (82.5%), gentamicin (81%), piperacillin/tazobactam (81%), cefepime (79%) and cefotaxime (%79%). All of the E. coli isolates which were resistant to ciprofloxacin and levofloxacin (44% and 43%, respectively) were found susceptible to fosfomycin tromethamine. In conclusion, since E. coli is by far the most prevalent community acquired urinary tract pathogen, fosfomycin tromethamine seems to be a reasonable alternative for the ampirical therapy of uncomplicated urinary tract infections. PMID- 17427561 TI - [Short communication: determination of hepatitis C virus genotypes by INNO-LIPA and sequence analysis methods]. AB - The detection of genotypes of hepatitis C virus (HCV) which exhibit very high genetic variability, has a great impact for the therapy and follow-up of the chronicity of infections. The aim of this study was to detect the genotypes of HCV strains by using two different methods. Thirty patients (5 hemodialysis patients, 9 chronic hepatitis C patients, 5 blood donors, 1 hospital staff) who were positive for both anti-HCV (Vitros, Ortho-Clinical Diagnostics) and HCV-RNA (Rotorgene, Artus) were included to the study. The serum samples were studied by Inno-LIPA (Inno-LIPA HCV-II, Innogenetics, Belgium) and sequence analysis (9700 Sequence Detection System, and ABI PRISM 310 Genetic Analyzer, Applied Biosystems, USA) methods. For Inno-LIPA, 5'non-coding region (5'NCR) of HCV-RNA was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) and genotyped by line probes. For sequence analysis (SA), NS5B and 5'NCR regions were amplified by RT-PCR, and genotypic variations were assessed by Cycle Sequencing system (Applied Biosystems, USA). As a result, one strain was found as 1a, and 28 strains were found as 1b with both Inno-LIPA and SA methods, however, one strain was genotyped as 1b/3a by Inno-LIPA, but as 1b by SA method. Our data have indicated that the results obtained by Inno-LIPA and sequence analysis methods were in concordance for the detection of HCV genotypes, considering that they have similar sensitivities. PMID- 17427562 TI - [Short communication: investigation of intestinal coccidia in patients with diarrhea]. AB - The intestinal protozoa have been increasingly identified in immunocompromised patients. In this study, stool samples of 554 patients [288 male, 266 female; 11 (2%) of them were immunocompromised] with diarrhea were examined between June 2004 and June 2005 in the Parasitology Laboratory of Dokuz Eylul University Hospital, Izmir, Turkey, in order to investigate the prevalence of intestinal coccidia. After formalin-ethyl acetate sedimentation procedure, the samples were examined by both native lugol and Kinyoun acid-fast staining methods for the detection of Cryptosporidium, Cyclospora and Isospora spp. As a result, Isospora belli oocysts were detected in the stool samples of two patients (0.4%) of which one was HIV positive and the other had experienced liver transplantation. Seven (1.3%) and two (0.4%) of the immunocompetent subjects were found positive for Cryptosporidium spp. and for Cyclospora cayetanensis, respectively. In conclusion, Isospora is one of the important protozoa which should be taken into consideration by clinicians in the immunocompromised patients with diarrhea. PMID- 17427563 TI - [A case of urinary tract infection caused by Flavimonas oryzihabitans]. AB - Flavimonas oryzihabitans is an uncommon bacterial species isolated from clinical specimens. In this report, a 53 years old female patient who had been followed up with the diagnosis of rectum cancer and renal failure for five and two years, respectively, was presented. F. oryzihabitans was isolated from the urine culture of the patient, and the clinical response to ofloxacin therapy was excellent even the urinary catheter has not been removed. Although this pathogen is mostly community aquired, it was found resistant to most of the antimicrobial agents tested. Since it was susceptible to fluoroquinolones and carbapenems, these antibiotics could be the drug of choice for the treatment of infections caused by F. oryzihabitans. The aim of this presentation was to withdraw attention to this bacterium which is thought to be the first urine isolate in our country. PMID- 17427564 TI - [Case report: tuberculosis of parotid gland]. AB - Primary tuberculosis of the parotid gland is an unusual clinical presentation. In this report a 32 years old male patient with parotid gland tuberculosis has been presented. The patient has been admitted to Ear, Nose & Throat outpatient clinic of our hospital with the complaint of left facial nodule. Histopathologic examination of the needle aspiration biopsy (NAB) specimen yielded benign necrotic lenfoid tissue, and in cervical ultrasonography cystic formations in left parotid gland were detected. Since the nodule size has increased in the follow-up period, cervical tomography was performed and heterogenous mass in the left side with lobular contour and hypodense appearance in posterior cervical region was detected. Histopathologic examination of the repeated NAB revealed chronic sialadenitis and benign lymphoid hyperplasia, and the patient has undergone left parotidectomy and lymph node dissection. Histopathologic examination of the excisional biopsy specimen revealed necrotising granuloma with diffuse caseification lesions concordant with tuberculosis. The patient was diagnosed as parotid gland tuberculosis, and anti-tuberculous therapy was started with isoniazid, rifampin, ethambutol and pyrazinamide. The history of the patient pointed out that he had used steroid for four months with a suspective diagnosis of rheumatoid arthritis, and his father had tuberculosis. In conclusion, since tuberculosis is a common infection in our country, it should be considered in the differential diagnosis of parotid nodules. PMID- 17427565 TI - [A severe case of Weil's disease]. AB - Leptospirosis which is caused by Leptospira species, may present with clinical features that vary from a mild flu-like illness to an acute life-threatening condition. Weil's disease, the most severe form of leptospirosis is characterized by multiorgan involvement including liver, kidney and lungs. In this report a severe Weil's disease was presented. A 43 years old male patient who had a history of swallowing water while swimming in the creek, was admitted to the hospital with the complaints of weakness, cough, bloody sputum, generalized jaundice and dark urine. Acute renal failure, bilateral lung infiltration, hyperbilirubinemia, leukocytosis and thrombocytopenia were detected, and the patient has undergone to hemodialysis. Ceftriaxone and ciprofloxacin treatment was applied to the patient after collection of blood, urine and sputum cultures and serum samples for serological tests. None of the cultures yielded pathogenic microorganisms. Microscopic agglutination test (MAT) was applied to two serum samples which were collected with 10 days interval. The first serum sample revealed antibody positivity at 1/200 titer against L. semeranga Patoc I, while the second serum revealed antibody positivity at 1/400 titer against both L. semeranga Patoc I and L. icterohaemorrhagiae Wijnberg. By the administration of antibiotic therapy and early supportive care the patient was cured completely. In conclusion Weil's disease should be taken into consideration in the patients with multiple organ involvements. PMID- 17427566 TI - [A case of brucellosis presenting as high titer negative result by standard tube agglutination test]. AB - Since prozone is a well known phenomenon in the serologic diagnosis of Brucella infections, it is necessary to prepare higher serum dilutions in the standard tube agglutination (STA) test for the brucellosis suspected patients. However, due to limited economical support, the serum dilutions generally last at 1/320 1/640 titers in some laboratories in Turkey. In this report, a brucellosis case whose STA test was found negative until the titer of 1/1280, has been presented. A 36-year-old female was admitted to our hospital with the complaints of fever, sweating, fatigue, generalized arthralgia and weight loss, lasting for 45 days. Hepatosplenomegaly was detected in the physical examination, and laboratory tests yielded anemia, leucopenia, elevated erythrocyte sedimentation rate and high C reactive protein levels. Although brucellosis was suspected, Brucella STA test was found negative at 1/640 titer. On the sixth day of admission, Brucella melitensis was isolated from her blood culture. Since a positive result at 1/40 titer was detected in Brucella STA test with the use of Coombs antiserum, the patient's serum was retested at higher dilutions than 1/640, and positive result was obtained starting from 1/1280 dilution and extended to 1/5120 titer. The patient was treated with rifampin and doxycyline and discharged with complete cure. In conclusion, in countries endemic for brucellosis, STA test should be performed at 1/1280 or higher titrations in suspected patients especially in the presence of negative culture results, for the prevention of false negative results due to prozone phenomenon. PMID- 17427567 TI - [Bacterial resistance to antiseptics and disinfectants]. AB - Antiseptic and disinfectant products contain active chemical agents known as biocides. The responses of different types of microorganisms to antiseptics and disinfectants vary and result in microbiostatic or microbicidal effects. Antiseptic and disinfectant resistance can be intrinsic or acquired by mutation or acquisition of plasmids or transposons. The mechanisms of resistance to antiseptics and disinfectants include cellular impermeability, biofilm formation, efflux and mutation(s) at the target site or overexpression of the target. It is important to know the mode of action of antiseptics and disinfectants and the mechanisms of microbial resistance in order to provide more rational use and production of more effective compounds. In this review article, bacterial resistance mechanisms to antiseptics and disinfectants have been discussed. PMID- 17427568 TI - [HbsAg seroprevalence in a county in Turkey where tobacco labours are prevalent]. PMID- 17427569 TI - Call for emergency networks to improve children's care. PMID- 17427570 TI - New guidelines for managing patients with burns. PMID- 17427571 TI - Responsible behaviour. PMID- 17427572 TI - Pain: a multifaceted phenomenon. PMID- 17427573 TI - The right to refuse. PMID- 17427574 TI - Staphylococcal scalded skin syndrome. PMID- 17427575 TI - Endotracheal tubes: early detection of oesophageal intubation. PMID- 17427576 TI - The emergency care nurse role: a clinical staff attitude survey. PMID- 17427577 TI - Board's eye view. PMID- 17427578 TI - Keep your eye on the prize. PMID- 17427579 TI - Social networking. PMID- 17427581 TI - Meet Dr Carl Tinkelman, "zoo-a-dontist". PMID- 17427580 TI - Governor signs "Cover All Kids" legislation. PMID- 17427582 TI - Pregnant women in your practice--What do we know? What should we do? PMID- 17427583 TI - A quantum leap in dental technology. PMID- 17427584 TI - "Tell someone" vaccine first new option available for prevention of cervical cancer. PMID- 17427585 TI - Miss communication. PMID- 17427586 TI - The EMTALA TAG. PMID- 17427587 TI - Surgery's 2007 federal advocacy agenda. PMID- 17427588 TI - Staying safe: simple tools for safe surgery. PMID- 17427589 TI - Retired surgeon is now a "clock doctor". PMID- 17427591 TI - Statement on surgery using lasers, pulsed light, radiofrequency devices, or other techniques. PMID- 17427590 TI - The economics of managed care reimbursement: a rationale for nonparticipation. PMID- 17427592 TI - Getting ready for Medicare's new quality reporting program. PMID- 17427593 TI - Access to emergency care. PMID- 17427594 TI - Access to emergency care. PMID- 17427595 TI - October Bulletin. What surgeons should know about...trends in Medicare reimbursement. PMID- 17427596 TI - Reimbursement. PMID- 17427597 TI - Does socio-economic status influence the use of mental health services? PMID- 17427598 TI - Disentangling associations between poverty at various levels of aggregation and mental health. AB - The present editorial discusses whether socioeconomic status of the individual and of the neighbourhood could be important in prevalence, treatment and prevention of psychiatric morbidity. Previous research showed that patients diagnosed with mental disorders are concentrated in socioeconomically disadvantaged areas. This could be the result of (1) an association between individual socioeconomic status and mental health, (2) an association between neighbourhood socioeconomic status and mental health, or (3) social selection. Research disentangling associations between individual and neighbourhood socioeconomic status on the one hand and mental health outcomes on the other, reported that neighbourhood socioeconomic disadvantage was associated with individual mental health over and above individual-level socioeconomic status, indicating deleterious effects for all inhabitants both poor and affluent. In conclusion, subjective mental health outcomes showed stronger evidence for an effect of neighbourhood socioeconomic status than research focussing on treated incidence. Within the group of patients, however, service use was higher in patients living in disadvantaged neighbourhoods. Social capital was identified as one of the mechanisms whereby neighbourhood socioeconomic disadvantage may become associated with observed reductions in mental health. After controlling for individual socioeconomic status, there is evidence for an association between neighbourhood socioeconomic status and objective as well as subjective mental health in adults. Evidence for such an association in young children is even stronger. PMID- 17427599 TI - Socio-economic status and geographies of psychiatric inpatient service use. Places, provision, power and well-being. AB - This editorial briefly summarises some aspects of research on socio-economic status and use of mental health services that have particular relevance for the theme of this issue of Epidemiologia e Psichiatria Sociale. This discussion takes a view from the perspective of health geography, which examines how the relationships between individuals and their social and physical environment result in variations in health and health care use. Three particular issues are considered here. First, the geographical distribution and organisation of psychiatric services may interact with social and economic factors in ways that are important for service use. Second, increasingly sophisticated ecological modelling strategies have elucidated the associations between socio-economic factors and service use at the population level. Third, more intensive, qualitative research complements these statistical analyses and encouraged reflection on the socio-economic processes, within psychiatric care settings, as well as in wider society, which influence service use. PMID- 17427600 TI - What is the impact of socio-economic inequalities on the use of mental health services? AB - Amartya Sen, who received the Nobel Prize for Economics, has demonstrated that the incidence of deprivation, in terms of capability, can be surprisingly high even in the most developed countries of the world. The study of socio-economic inequalities, in relation to the utilisation of health services, is a priority for epidemiological research. Socio-economic status (SES) has no universal definition. Within the international research literature, SES has been related to social class, social position, occupational status, educational attainment, income, wealth and standard of living. Existing research studies have shown that people from a more deprived social background, with a lower SES, are more likely to have a higher psychiatric morbidity. Many studies show that SES influences psychiatric services utilization, however the real factors linking SES and mental health services utilisation remain unclear. In this editorial we discuss what is currently known about the relationship between SES and the use of mental health services. We also make an argument for why we believe there is still much to uncover in this field, to understand fully how individuals are influenced by their personal socio-economic status, or the neighbourhood in which they live, in terms of their use of mental health services. Further research in this area will help clarify what interventions are required to provide greater equality in access to mental health services. PMID- 17427601 TI - What is a risk ratio? PMID- 17427602 TI - Psychoeducational family interventions for schizophrenia in the last decade: from explanatory to pragmatic trials. AB - A number of explanatory RCT studies published since the 1980s have demonstrated the clinical efficacy of Psychoeducational Family Interventions (PFI) for schizophrenia when provided in combination with drug therapy. In recent years, there has been a shift from efficacy to effectiveness studies and great attention by the researchers in developing training programmes in these interventions for ordinary staff. In this paper, we will provide an overview of the studies on PFI for schizophrenia which have been carried out in the last decade in routine clinical settings or with at least a partial involvement of ordinary staff. These studies have been grouped into: (a) studies comparing PFI with standard care; (b) studies comparing PFI with individual integrated interventions; (c) studies comparing different PFI strategies; (d) implementation studies. The results of these studies reveal that, when provided in clinical settings, PFI have positive middle-term effects on patients' clinical status and disability, and limited impact on family burden. From a methodological viewpoint, these studies had several similarities, such as homogeneity of PFI models and mid-term follow-up assessments, and several differences, mainly in the intensity and duration of the family exposure to the intervention. Future studies are needed to identify the "best dose" at which PFI can be provided in routine conditions at the most convenient cost-benefit ratio. PMID- 17427603 TI - [The diffusion and the reason for the use of complementary and alternative medicine among users of mental health services: a systematic review of literature]. AB - AIMS: A systematic review of the studies on use of alternative and complementary medicine by mental health services' users to investigate the prevalence and motivations of CAM used to treat mental disorders. METHODS: This paper shows a part of the results of a literature review, limited to studies between 1966 and 2005, was carried out using Ovid Medline, PsycINFO and EBM Reviews databases. This was supplemented by hand searching of reference lists. RESULTS: Ninety-one studies on CAM's use by psychiatric patients were analyzed. In this paper the twenty-five articles about the CAM's prevalence and use's motivations are described. The use of complementary and alternative medicine (CAM) varies across countries: data suggest that from 50% to 80% of people suffering of mental disorders use CAM. Heterogeneous methods, samples and results characterizing the studies make generalizations results difficult. Patients motivations of CAM's use were poorly investigated by research, however it seems that persons affected by mental disorders use MAC prevalently because they are dissatisfied with conventional medicine and/or because these alternatives approaches are congruent with their own values, beliefs and philosophical orientation toward health care. CONCLUSIONS: The review emphasize the importance of a best physicians' education and information about CAM to improve the management of non-conventional and orthodox medicine relationship. The motivations of CAM's use underline the relevance of patient's holistic conception and the therapeutic role of a patient physician relationship based on emphaty. Besides it seems important to investigate furtherly patients' psychological and social aspects that influence their choise to use CAM. PMID- 17427604 TI - Relapsing versus non relapsing course of schizophrenia: a cohort study in a community based mental health service. AB - AIM: We examined the effect of several clinical variables on the tendency to relapse and to require hospitalization in a cohort of patients, living in the community and followed up naturalistically for seven years. METHOD: Forty-six patients affected by schizophrenia and schizoaffective disorder, according to both DSM-IV and ICD-10 criteria, were assessed by Positive and Negative Syndrome Scale and Life Skills Profile (LSP). All patients consecutively enrolled, were assessed in a stable clinical phase of illness and treated as usual by their reference psychiatrist. Social and clinical outcome was assessed yearly for seven years after the study entry and analyzed with survival analysis. RESULTS: Patients who did not relapse, were characterized by higher functioning, lower positive symptoms, higher ability in self-care and non-turbulence and higher IQ at their baseline clinical evaluation. These variables were entered in a Cox regression model to corroborate the predictive power on the relapsing course of illness. Only IQ and non-turbulence scores of LSP were entered in the equation (Wald method: p = 0.007 and p = 0.002 respectively). CONCLUSIONS: Several factors interact with the course of illness and influence the tendency to require hospitalization. In the present study we report that non-turbulence is a significant predictor of a non-relapsing course of illness. Further studies are needed to clarify the role of other mediating variables. PMID- 17427605 TI - [Heterogeneity of the Departments of Mental Health in the Veneto Region ten years after the National Plan 1994-96 for Mental Health. Which implication for clinical practice? Findings from the PICOS Project]. AB - AIMS: This study aims to present data on structural and human resources of public mental health services located in the Veneto Region, Italy, and to discuss them in the light of implementation of the first National Target Plan for Mental Health ("Progetto Obiettivo 1994-1996") ten years after its launch. METHODS: The study was conducted in the context of the PICOS (Psychosis Incident Cohort Outcome Study) Project, a large first-presentation multisite study on patients with psychotic disorders attending community mental heath services in the Veneto Region. Human and structural resources were surveyed in 26 study sites using a structured interview administered by the PICOS local referents. RESULTS: CMHCs and Day Centres were homogeneously distributed across the Region and their overall rates per resident population met the national standards; a wide variability in the distribution of Day Hospitals was found, with the overall rate per resident population very far from meeting the national standard; the overall rate for Residential Facilities beds was higher than the recommended national standard, showing however an high variability across sites. The overall rate of mental health professionals per resident population was only slightly below the national standard: this was mainly achieved thanks to non-profit organizations which supplement the public system with unspecialised professionals; however, a wide variability in the local rates per resident population was found, with the 50% of the sites showing rates far lower the national standard. Specific lack of trained professionals involved in the provision of psychosocial interventions was found in most sites. CONCLUSIONS: A marked variability in human and structural resources across community mental health services in the Veneto Region was found. Possible reasons for this heterogeneity were analysed and implications for mental health care provision were further discussed. PMID- 17427606 TI - Implications of changes in the impact factors of psychiatric journals. PMID- 17427607 TI - [Substance use and psychiatric symptoms in subjects referred to the drug addiction services by the prefecture]. PMID- 17427608 TI - [Validity and reliability of Quality of Life, Enjoyment and Satisfaction Questionnaire, Short Form]. PMID- 17427609 TI - Management of diabetes should be a team approach. PMID- 17427610 TI - The preparticipation evaluation: evolving to enhance the health and safety of athletes. PMID- 17427611 TI - Cardiovascular disease prevention in women. PMID- 17427612 TI - Are opioids effective in the treatment of neuropathic pain? PMID- 17427613 TI - Decongestants and antihistamines do not relieve symptoms of otitis media with effusion. PMID- 17427614 TI - The athletic preparticipation evaluation: cardiovascular assessment. AB - Thousands of young athletes receive preparticipation evaluations each year in the United States. One objective of these evaluations is to detect underlying cardiovascular abnormalities that may predispose an athlete to sudden death. The leading cardiovascular causes of sudden death in young athletes include hypertrophic cardiomyopathy, congenital coronary artery anomalies, repolarization abnormalities, and Marfan syndrome. Because these abnormalities are rare and difficult to detect clinically, it is recommended that family physicians use standardized history questions and examination techniques. Athletes, accompanied by their parents, if possible, should be asked about family history of cardiac disease and sudden death; personal cardiac history; and exercise-related symptoms, specifically syncope, chest pain, and palpitations. The physical examination should include blood pressure measurement, palpation of radial and femoral pulses, dynamic cardiac auscultation, and evaluation for Marfan syndrome. Athletes with "red flag" signs or symptoms may need activity restriction, special testing, and referral if the diagnosis is unclear. PMID- 17427615 TI - Work-related eye injuries and illnesses. AB - More than 65,000 work-related eye injuries and illnesses, causing significant morbidity and disability, are reported in the United States annually. A well equipped eye tray includes fluorescein dye, materials for irrigation and foreign body removal, a short-acting mydriatic agent, and topical anesthetics and antibiotics. The tray should be prepared in advance in case of an eye injury. Eye patching does not improve cornea reepithelialization or discomfort from corneal abrasions. Blunt trauma to the eye from a heavy object can cause a blow-out fracture. Sudden eye pain after working with a chisel, hammer, grinding wheel, or saw suggests a penetrating globe injury. Chemical eye burns require immediate copious irrigation. Nontraumatic causes of ocular illness are underreported; work related allergic conjunctivitis increasingly has been recognized among food handlers and agriculture workers who are exposed to common spices, fruits, and vegetables. The patient's history of eye injury guides the diagnosis. Primary prevention and patient counseling on proper eye protection is essential because over 90 percent of injuries can be avoided with the use of eye protection. As laser use increases in industry and medical settings, adequate personal protection is needed to prevent cataracts. Outdoor workers exposed to significant ultraviolet rays need sun protection and safety counseling to prevent age-related macular degeneration. Contact lenses do not provide eye protection, and physicians should be familiar with guidelines for the use of contacts in the workplace. PMID- 17427616 TI - Information from your family doctor. Work-related eye injuries and illnesses: what you should know. PMID- 17427617 TI - Peppermint oil. AB - Peppermint leaf and peppermint oil have a long history of use for digestive disorders. Recent evidence suggests that enteric-coated peppermint oil may be effective in relieving some of the symptoms of irritable bowel syndrome. A combination product including peppermint oil and caraway oil seems to be moderately effective in the treatment of non-ulcer dyspepsia. Topical application of peppermint oil may be effective in the treatment of tension headache. Because of its relaxing effects on smooth muscle, peppermint oil given via enema has been modestly effective for relief of colonic spasm in patients undergoing barium enemas. Peppermint oil is well tolerated at the commonly recommended dosage, but it may cause significant adverse effects at higher dosages. PMID- 17427618 TI - Outpatient management of anticoagulation therapy. AB - The Seventh American College of Chest Physicians (ACCP) Conference on Antithrombotic and Thrombolytic Therapy provides guidelines for outpatient management of anticoagulation therapy. The ACCP guidelines recommend short-term warfarin therapy, with the goal of maintaining an International Normalized Ratio (INR) of 2.5 +/- 0.5, after major orthopedic surgery. Therapy for venous thromboembolism includes an INR of 2.5 +/- 0.5, with the length of therapy determined by associated conditions. For patients with atrial fibrillation, the INR is maintained at 2.5 +/- 0.5 indefinitely; for most patients with mechanical valves, the recommended INR is 3.0 +/- 0.5 indefinitely. Use of outpatient low molecular-weight heparin (LMWH) is as safe and effective as inpatient unfractionated heparin for treatment of venous thromboembolism. The ACCP recommends starting warfarin with unfractionated heparin or LMWH for at least five days and continuing until a therapeutic INR is achieved. Because patients with venous thromboembolism and cancer who have been treated with LMWH have a survival advantage that extends beyond their venous thromboembolism treatment, the ACCP recommends beginning their therapy with three to six months of LMWH. When invasive procedures require the interruption of oral anticoagulation therapy, recommendations for bridge therapy are determined by balancing the risk of bleeding against the risk of thromboembolism. Patients at higher risk of thromboembolization should stop warfarin therapy four to five days before surgery and start LMWH or unfractionated heparin two to three days before surgery. PMID- 17427619 TI - Parkinson's disease. PMID- 17427620 TI - FPIN's clinical inquiries. Glycemic control in patients with type 2 diabetes. PMID- 17427621 TI - An infant with a hypopigmented macule. PMID- 17427622 TI - Physicians as role models. PMID- 17427623 TI - Bigger, yes, but better? UnitedHealth Group's planned acquisition of Sierra Health leaves some wary of a trend that could disrupt service, payment. AB - "Oh no. Not again." That was one CEO's reaction to UnitedHealth's deal for Sierra, Nevada's top health insurer. Recent takeovers, including the WellPoint/Anthem deal, have left bad memories. "Since then, the race downward on payments has been dramatic, so you can see why these kinds of mergers create a lot of anxiety", says Lawrence Matheis, left. PMID- 17427624 TI - Satisfied customers. Survey gives not-for-profits good grades. PMID- 17427625 TI - Bush won't budge. Citizens' group report on improving coverage rejected. PMID- 17427626 TI - Congress works to save SCHIP. Hospitals don't want to be targeted to offset new funds. PMID- 17427627 TI - Access varies with income: JAMA. Study shows many patients squeezed on paying for care. PMID- 17427628 TI - Cleveland Clinic in another deal. Proposed telemedicine partnership raises conflict-of-interest concerns. PMID- 17427629 TI - Friends go to bat for AHRQ. PMID- 17427630 TI - Fostering ethics. How developing individual, group expectations make for a better organization. PMID- 17427631 TI - Health care hall of fame. Keeping her word. Brinker's breast cancer foundation started 25 years ago with a sisterly promise. PMID- 17427632 TI - Health care hall of fame. Visionary founder. Cardwell helped pioneer School of Health Administration at Va. University. PMID- 17427633 TI - Health care hall of fame. Never alone at the wheel. Warden's team-oriented leadership drove Ford's growth over 15-year tenure. PMID- 17427634 TI - The end of 'fair share'? With Maryland law struck down, states work to protect coverage mandates. PMID- 17427635 TI - Help from the outside. States use variety of contractors to provide healthcare for prisoner population. PMID- 17427636 TI - By the numbers. Largest behavioral health providers. Ranked by behavioral health net patient revenue. PMID- 17427637 TI - The usefulness of angiotensin converting enzyme in the differential diagnosis of Crohn's disease and intestinal tuberculosis. AB - BACKGROUND: Since the pathologic findings of Crohn's disease (CD) and intestinal tuberculosis (IT) overlap to a large degree, the development of other biomarkers will be of great help for making the differential diagnosis of these 2 diseases. The aim of the present study is to examine the clinical efficacy of using the tissue angiotensin converting enzyme (ACE) assay in making the differential diagnosis between CD and IT. METHODS: Tissue specimens were obtained from 36 patients who were diagnosed with CD or IT by the colonoscopic biopsy, as well as by the clinical findings. The expression of tissue ACE was detected by immunohistochemical staining. The optimal cut-off value of the immunoreactive scoring (IRS) system we used to differentiate CD from IT was determined by analysis of the ROC curve and AUROC. RESULTS: Granuloma was present in 15 of 19 patients with CD (78.9%) and in 15 of 17 patients with IT (88.2%). ACE was present in the cytoplasm of the epithelioid cells in the granulomas from 13 of 15 patients with CD and in 14 of 15 patients with IT. The IRS scores of ACE were greater in the patients with CD than that of the patients with IT (8.07 +/- 4.38 vs. 4.13 +/- 2.47, respectively, p = 0.006). In differentiating CD from IT, the AUROC curve for the IRS of ACE was 0.767 with a sensitivity of 66.7%, a specificity of 93.3% and the cut-off point was 7.5. CONCLUSIONS: The results of our study suggest that the assessment of the tissue ACE expression can be helpful for making the differential diagnosis between CD and IT. PMID- 17427638 TI - Volatile organic compounds contribute to airway hyperresponsiveness. AB - BACKGROUND: Volatile organic compounds (VOCs) in concentrations found in both the work and home environments may influence lung function. We investigated the prevalence of airway responsiveness in workers exposed to VOCs. METHODS: We used allergic skin tests, nonspecific airway hyperresponsiveness testing and questionnaires to study twenty exposed workers and twenty-seven control subjects. Atopy was defined as a reactor who showed > 3+ response to one or more allergens on the skin prick tests. Airway hyperresponsiveness (BRindex) was defined as log [% fall of FEV1/log (last concentration of methacholine) +10]. RESULTS: The VOC exposed workers, in comparison with the control subjects, tended to have a higher BRindex (1.19 +/- 0.07 vs. 1.15 +/- 0.08, respectively). Workers exposed to VOCs with atopy or smoker, as compared with the workers exposed to VOCs with non-atopy and who were non-smokers and the control subjects with non-atopy and who were non smokers, had a significantly higher BRindex (1.20 +/- 0.05 vs. 1.14 +/- 0.06 vs. 1.10 +/- 0.03, respectively p < 0.05). The BRindex was not correlated with atopy, the smoking status or the duration of VOC exposure. CONCLUSIONS: These findings suggest that VOCs may act as a contributing factor of airway hyperresponsiveness in workers exposed to VOCs. PMID- 17427639 TI - Eosinophilic enteritis presenting as intussusception in adult. AB - Eosinophilic gastroenteritis is defined as a disorder that selectively affects the gastrointestinal tract with eosinophil-rich inflammation in the absence of any known causes for eosinophilia. The clinical manifestations vary according to the site of the eosinophilic infiltrated layer of the bowel wall. Eosinophilic enteritis presenting as intussusception in adult has not been previously reported in the literature. Especially, making the diagnosis of intussusception in adults is often difficult due to the variable clinical findings. In our case, the correct diagnosis of intussusception due to eosinophilic enteritis was arrived at rather easily based on the ultrasonography and endoscopic biopsy. The patient was treated with oral prednisolone at 30 mg/day for 7 days, and then the drug was tapered off for 2 months; we didn't perform surgery. He has been asymptomatic for about 1 year after discharge without disease recurrence. PMID- 17427640 TI - Adrenalectomy for metastatic disease to the adrenal gland from gastric cancer: report of a case. AB - Metastases to the adrenal glands are frequently found at autopsy. In practice, adrenal metastases have generally been accepted as evidence of blood-borne systemic disease. So, clinically curable adrenal metastases is a rare malady. The role for surgical resection in adrenal metastases has not been clearly defined. A 45-year-old man initially underwent total gastrectomy with D2 lymph node dissection for treating his advanced gastric cancer. A solitary adrenal metastases was resected 1 year later. The patient has survived for 3 years and no further evidence of disease was found on his last follow-up examination. We report here on this case to show that for selected cases, surgical resection of adrenal metastases is feasible and this procedure may extend survival for metastatic gastric cancer patients. PMID- 17427641 TI - Malignant gastrointestinal stromal tumor in a patient with neurofibromatosis type 1. AB - Neurofibromatosis type 1 (von Recklinghausen's disease, NF-1) is an autosomal dominant neurocutaneous disorder characterized by abnormal skin pigmentation (cafe au lait spots and axillary freckling), cutaneous and plexiform neurofibromas, skeletal dysplasias, and Lisch nodules (pigmented iris hamartomas). Gastrointestinal stromal tumors (GISTs) are the most common tumors of mesenchymal origin in the gastrointestinal tract, mesentery, omentum, and retroperitoneum. Here, we report a case of GIST in the ileum of a 76-year-old woman previously diagnosed as NF-1. She was admitted due to sudden onset of abdominal pain. Contrast enhanced CT scan revealed a moderately defined, peripherally enhanced soft tissue mass of about 8.8 x 7.3cm, originating from the small bowel in the left of the abdomen. Surgical excision was performed and the tumor was found to be composed of tumor cells that were positive for c-kit protein. The patient started imatinib treatment a month later, but stopped medication due to dyspepsia after a few months and eventually progressed after 18 months. PMID- 17427642 TI - Remission of intestinal Behcet's disease treated with anti-tumor necrosis factor alpha monoclonal antibody (Infliximab). AB - Behcet's disease (BD) is a chronic relapsing multisystem disease characterized by oral ulceration, genital ulceration and ocular lesions. Gastrointestinal involvement is rare, often difficult to treat and associated with a high mortality rate. We treated a 47-year-old Korean man with BD who had a recurrent intestinal ulcer with tumor necrosis factor alpha antibody (infliximab); he initially underwent right hemicolectomy due to uncontrolled intestinal bleeding. For patients with intestinal BD who fail to respond to conventional treatment, infliximab may be a safe and effective new therapeutic option. PMID- 17427643 TI - Hypertrophic cardiomyopathy complicated by left ventricular apical necrosis and aneurysm in a young man: FDG-PET findings. AB - A 29-year old male was transferred to our hospital with an abnormal chest X-ray finding diagnosed as hypertrophic cardiomyopathy with apical necrosis and aneurysm formation. Four years after the initial hospitalization, we confirmed the aneurysm and necrosis using both integrated positron emission tomography (PET) and computed tomography (CT) scanning. The F-18 2-fluoro-2-deoxy-D-glucose (FDG) PET/CT enabled precise localization of the aneurysm, which was found to be composed of semi-lunar calcification of non-metabolic myocardium. A contrast enhanced CT angiography showed an hour-glass appearance of the left ventricular cavity. The integrated PET/CT fusion scanner is a novel multimodality technology that allows for a comprehensive analysis of the anatomical and functional status of complex heart disease. Based on these findings, long standing mechanical and physiologic abnormalities may have led to chronic ischemia in the hypertrophied myocardium, induced necrosis and calcification at the cardiac apex. PMID- 17427644 TI - A case of pulmonary vein tumor presenting as a left atrial mass. AB - Primary cardiac tumors are extremely rare and can originate within the heart or be the result of tumor spread from other sites. We report a female patient with a pulmonary vein tumor extending into the left atrium that had a suspicious primary malignant origin with a sacral metastatic carcinoma. The patient was admitted complaining of pain in her buttock area as a result of a sacral tumor. It was believed that the sacral tumor was a metastasis from the imaging study and clinical manifestation. The primary malignant origin was evaluated. The chest CT showed a left atrium thrombus-like lesion without a pulmonary abnormality. After a transesophageal echocardiogram, the patient was diagnosed with a pulmonary vein tumor extending to the left atrium. The patient was given palliative radiotherapy for the sacral pain. Initially, the clinical impression was a metastatic sacral tumor with a thromboembolism of the left atrium. However, this patient was finally diagnosed with a pulmonary vein tumor with a left atrium extension by a transesophageal echocardiogram. PMID- 17427645 TI - Use of tadalafil for treating pulmonary arterial hypertension secondary to chronic obstructive pulmonary disease. AB - Pulmonary arterial hypertension (PAH) secondary to chronic obstructive pulmonary disease (COPD) is incurable and it has an unpredictable survival rate. Two men who suffered from COPD presented with progressive dyspnea and edema, respectively. PAH, as estimated by the peak velocity of tricuspidal regurgitation, and the depressed myocardial performance index (MPI) of the right ventricle (RV) were noted on echocardiography. In addition to the baseline therapy for their depressed ventilatory function, we prescribed tadalafil 10 mg orally every other day for 2 weeks and then we doubled the dosage. They well tolerated the medication without any notable side effects. After 4 weeks of tadalafil treatment, the patients' pulmonary arterial pressure was decreased and the MPI of the RV was improved in both. The exercise capacity, as measured by the respiratory oxygen uptake, also improved from 10.9 mL/kg/min to 13.8 mL/kg/min in one patient. We report here on 2 patients with PAH secondary to COPD, and they showed notable improvement of their pulmonary hemodynamics and exercise capacity with the administration of tadalafil. PMID- 17427646 TI - Pneumatosis intestinalis with pneumoperitoneum mimicking intestinal perforation in a patient with myelodysplastic syndrome after hematopoietic stem cell transplantation. AB - Pneumatosis intestinalis (PI) is an uncommon disorder characterized by an accumulation of gas in the bowel wall, and has been associated with a variety of disorders and procedures. We describe a 35-year-old man who undertook hematopoietic stem cell transplantation due to myelodysplastic syndrome. An abdominal X-ray demonstrated extensive PI with pneumoperitoneum mimicking hollow organ perforation. However, the patient had no abdominal symptoms and there was no evidence of peritoneal inflammation. After two weeks of conservative management, including bowel rest and antibiotics, his pneumoperitoneum resolved spontaneously without any complications. Of the many factors that affect the gastrointestinal tract mucosal integrity, intramural pressure, and bacterial flora-produced intraluminal gas interact to produce PI. If the condition is accompanied by bowel ischemia, portomesenteric venous gas, metabolic acidosis, and abdominal sepsis, or if PI is severe in extent immediate surgical intervention is indicated. The described case supports that a mechanical rather than a bacterial etiology underlies the pathogenesis of PI. PMID- 17427647 TI - A case of McCune-Albright syndrome with associated multiple endocrinopathies. AB - McCune-Albright syndrome (MAS) is a rare disorder that develops from an activating mutation in the Gs gene. It is characterized by an association with Polyostotic fibrous dysplasia, and precocious puberty, Caf-au-lait pigmentation, and other endocrinopathies that result from the hyperactivity of a variety of endocrine glands. Recently we encountered a patient with MAS with fibrous dysplasia, skin pigmentation, acromegaly, hyperprolactinemia and a thyroid nodule. A 23-year-old male presented for an evaluation of a change in his facial structures. Fibrous dysplasia was diagnosed by a bone biopsy and radiographic studies. The GH level increased paradoxically after an oral glucose load. The plasma prolactin, IGF-1 and alkaline phosphatase were high. Thyroid ultrasonography revealed multiple nodules. The brain MRI demonstrated a mass in the left pituitary gland. Genetic analysis identified a change from Arg (CGT) at codon 201 to Cys (TGT). PMID- 17427648 TI - Paroxysmal autonomic dysregulation with fever that was controlled by propranolol in a brain neoplasm patient. AB - Intractable fever in cancer patients is problematic and the causes of this fever can be diverse. Paroxysmal persistent hyperthermia after sudden mental change or neurologic deficit can develop via autonomic dysregulation without infection or any other causes of fever. Paroxysmal hyperthermic autonomic dysregulation is a rare disease entity. It manifests as a form of paroxysmal hypertension, fever, tachycardia, tachypnea, pupillary dilation, agitation and extensor posturing after traumatic brain injury, hydrocephalus, brain hemorrhage or brain neoplasm. We recently experienced a case of paroxysmal hyperthermia following intracerebral hemorrhage along with brain neoplasm. Extensive fever workups failed to show an infectious or inflammatory source and/or hormonal abnormality. Empirical treatments with antibiotics, antipyretics, morphine, steroid and antiepileptic agents were also ineffective. However, Propranolol, a lipophilic beta-blocker, successfully controlled the fever and stabilized the patient. Fever in cancer patients is a common phenomenon, but a central origin should be considered when the fever is intractable. Propranolol is one of the most effective drugs for treating paroxysmal hyperthermia that is due to autonomic dysregulation. PMID- 17427649 TI - Acral metastasis in a patient with ampullary carcinoma. AB - Although skin metastasis from a malignant tumor of an internal organ usually occurs at an advanced disease stage, there has been no prior report of a cutaneous acral metastasis from ampullary carcinoma to date. We report a 71-year old male patient with cutaneous metastasis from an ampullary adenocarcinoma. The patient had a history of pylorus preserving pancreaticoduodenectomy for carcinoma of the ampulla of Vater two years prior to presentation. Physical examination revealed ill-defined, painful and hard erythematous nodules at the left thumb and distal phalanx of the right middle finger. The computed tomography scan showed low density masses in the retroperitoneum; the histological examination of a nodule from the right middle finger showed a metastatic adenocarcinoma. This case illustrates that cutaneous metastasis from ampullary carcinoma has a poor prognosis. PMID- 17427650 TI - Esophageal thermal injury by hot adlay tea. AB - Reversible thermal injury to the esophagus as the result of drinking hot liquids has been reported to generate alternating white and red linear mucosal bands, somewhat reminiscent of a candy cane. This phenomenon is associated with chest pain, dysphagia, odynophagia, and epigastric pain. Here, we report a case of thermal injury to the esophageal and oral cavity due to the drinking of hot tea, including odynophagia and dysphagia. A 69-year-old man was referred due to a difficulty in swallowing which had begun a week prior to referral. The patient, at the time of admission, was unable to swallow even liquids. He had recently suffered from hiccups, and had consumed five cups of hot adlay tea one week prior to admission, as a folk remedy for the hiccups. Upon physical examination, the patient's oral cavity evidenced mucosal erosion, hyperemia, and mucosa covered by a whitish pseudomembrane. Nonspecific findings were detected on the laboratory and radiological exams. Upper endoscopy revealed diffuse hyperemia, and erosions with thick and whitish pseudomembraneous mucosa on the entire esophagus. The stomach and duodenum appeared normal. We diagnosed the patient with thermal esophageal injury inflicted by the hot tea. He was treated with pantoprazole, 40 mg/day, for 14 days, and evidenced significant clinical and endoscopic improvement. PMID- 17427651 TI - Adenoviral pneumonia during etanercept treatment in a patient with rheumatoid arthritis. AB - Inhibitors of tumor necrosis factor-alpha (TNF-alpha) have been approved for treating rheumatoid arthritis. As one of the biological response modifiers, etanercept has also been used in the treatment of psoriatic arthritis and inflammatory bowel disease. While etanercept is effective, certain infectious complications, such as tuberculosis, fungus, and cytomegalovirus, have been reported. We report the first Korean case of adenoviral pneumonia in a 55-year old female who developed disseminated adenoviral infection following etanercept treatment, which resolved after anti-TNF-alpha discontinuation. PMID- 17427652 TI - Issues and answers: nursing value. PMID- 17427653 TI - Health policy report: collaboration--it's everywhere! PMID- 17427654 TI - Men in nursing: still a new frontier. PMID- 17427655 TI - Fathers: present, or just in the room? PMID- 17427656 TI - The Conservative view. PMID- 17427657 TI - Campaign for Normal Birth. Students' stories. PMID- 17427658 TI - Social enterprise: do midwives gain? PMID- 17427659 TI - Wellbeing and beyond. PMID- 17427660 TI - Free care for the displaced? PMID- 17427661 TI - Links to Ethiopia. PMID- 17427662 TI - A five-hour hike to give birth. PMID- 17427663 TI - What women want from postnatal care. PMID- 17427664 TI - The uniqueness of normality. PMID- 17427665 TI - Funding direction. PMID- 17427666 TI - [The death of father or mother leaves a painful void]. PMID- 17427667 TI - [The art of holistic care in pediatric oncology]. PMID- 17427668 TI - [Perioperative care of children with cleft lip-jaw-palate]. PMID- 17427669 TI - [Ambulatory tube weaning therapy for children with special needs]. PMID- 17427670 TI - [Pharmacotherapy of common colds in childhood]. PMID- 17427671 TI - [Travel into the light]. PMID- 17427672 TI - [A good start in life. New answers to new challenges]. PMID- 17427673 TI - [Evidence of a modification of the profile of addictive drugs consumption through the study of patients consulting a psychiatric emergency room]. AB - The consumption of psychoactive drugs is frequently the reason of consultation in a psychiatric emergency unit. We present here the evolution over time of the consumption pattern of patients consulting the psychiatric emergency unit of the Brugmann University Hospital. Major modifications were observed during this period. Mainly, problematic consumption of opiates was progressively overwhelmed by this of cannabis, psychostimulants and recently synthesis drugs. We suggest that this reflects the situation of psychoactive drugs consumption in the general population. This implicates a modification of prevention and treatment attitudes in drug consuming persons. PMID- 17427674 TI - [Neurophysiological and clinical aspects of psychophysiological insomnia]. AB - Nowadays, a high percentage of the population in modern societies suffers from various kinds of sleep disturbances, in part due to the impact of increased socio cultural stress factors. The most common sleep complaint is insomnia. Psychophysiological insomnia is one of the most frequent forms of sleep disturbances. It is a type of conditioned or learned insomnia that is presumably derived from two sources. The first involves the conditioning of sleep-preventing habits in which repeated pairing of sleeplessness and situational, temporal, or behavioural stimuli normally associated with sleep leads to conditional arousal that impairs sleep. The second involves somatised tension believed to result from the internalisation of psychological conflicts and excessive worrying about sleep, which are incompatible with sleep. Quantitative EEG approach, particularly with spectral analysis, has recently refined the analysis of the EEG in sleep. These EEG quantification techniques are now very useful to understand the neurophysiological basis of a lot of sleep disorders, particularly in the case of psychophysiological insomnia. PMID- 17427675 TI - [A medical reduced practice reimbursed by the sickness insurance is not synonymous with medical plethora in the French community of Belgium]. AB - OBJECTIVE: To determine if a reduced medical practice reimbursed by the sickness insurance is a proof of medical plethora. Enquiry design : Descriptive observation transverse design. POPULATION: The French-speaking general practitioners and specialists questioned in 2003 about their reduced activity volume recorded in 2000 by the National Institute of the Sickness - Disability insurance (INAMI) on the French Community territory. METHOD: All French-speaking general practitioners and specialists, not reaching the accreditation activity level in 2000, were selected starting from databank of the INAMI. A questionnaire was transmitted to the physicians by mailing via the INAMI services. The answer rate was 44,0 % for the specialists and 40,5 % for the general practitioners. RESULTS: In 2000, 47,7 % of the French-speaking general practitioners and 23,3 % of the French-speaking specialists did not reach the activity level given the right to be accredited by the INAMI. After the enquiry, only 5,5 % of the general practitioners and 5,7 % of the specialists had really a reduced activity. It was explained for a great part by the choice of career guidance out of the INAMI system and under-declaration of foreign working and retirement. CONCLUSIONS: The data bases of the INAMI are the only sources of individual medical activity recording. Their uses, in term of medical manpower planning, must be done with corrective factors issued from this study. The accreditation activity level is a good indicator to underline the activity out of the sickness insurance sector. PMID- 17427676 TI - [Fear of falling in older people]. AB - Fear of falling and poor physical performance are prominent symptoms in many older people. The prevalence of fear of falling in community-living elderly ranges between 12 % and 65 %, and is higher in women than men. It commonly occurs after falls, but it also occurs without a previous fall history. One of the major consequences of fear of falling is the restriction and avoidance of activities. However, not all elderly with fear of falling avoid activities in daily life. Some elderly only become cautious, which may be functional in preventing falls. Only a small percentage of elderly show a pattern of excessive fear and restriction of activities. The consequences of this pattern may, however, be debilitating and devastating. Excessive fear and avoidance may compromise the quality of life, and may result in a decline of physical capabilities and, ultimately, in an increased risk of falls, which may further fuel fear and avoidance. Future research should investigate whether individualised intervention strategies are efficient in preventing falls and activity-related fear of falling within this population in order to improve her quality of life. PMID- 17427677 TI - [The relevance of the concept of resiliency in the field of psychiatry]. AB - The concept of resilience refers to the capacity of certain individuals either to adapt successfully to adversity, or to function in a competent manner when faced with a traumatic situation. The concept of resilience represents a new paradigm in the field of clinical psychology. It puts into perspective determinist models, whether they be psychological (experiences incurred during the first years of life forever seal the destiny of a subject) or biological (an individual is defined by his/her genes). It permits the study of an individual according to his/her resources and no longer according to his/her faults. Nevertheless, for several years, resilience has suffered from its own success by being frequently used in inappropriate contexts. With the aim of better understanding this concept and of avoiding its determinist and normative distortion, we present here a review of the different factors (psychological, family, sociological and biological factors) contributing to the process of resilience. Moreover, in the discussion, we present some methodological and conceptual issues presented by the concept of resilience. PMID- 17427678 TI - [Placebo and placebo effects (second part) : ethical aspects]. AB - The use of placebo is controversial. It is generally accepted when no treatment exists for the disease under study but a polemic persists when a therapy is already regularly applied by medical practitioners. The paper cites and comments on the publications of national and international institutions (Belgian Medical Board, World Medical Association, Council for International Organizations of Medical Sciences) on this topic. The pros and cons about the use of placebo in clinical research are presented. Each protocol, with or without placebo has to abide by the general principles of medical ethics whose requirements may sometimes be contradictory. In these circumstances, the investigators must find a compromise acceptable morally and scientifically and submit it to ethics committees which have to decide about the validity of their arguments. PMID- 17427679 TI - [When a multiple trauma complains about abdominal pains six weeks after the fall. . ]. AB - A 18-year old patient presents abdominal pains associated with nausea and vomiting six weeks after a multiple trauma leading to paraplegia. Esophagitis, urolithiasis, and acalculous cholecystitis were diagnosed. This report illustrates two rare abdominal complications of a multiple trauma with immobilisation: acalculous cholecystitis, hypercalcaemia, coralliform urolithiasis following urinary infection and bacteriurie with Enterobacter Cloacae. The interpretation of the abdominal semiology of a multiple trauma's patient presenting long-term immobilization may be misleading. PMID- 17427680 TI - [Depression in children]. PMID- 17427681 TI - [Diffusion, stress of diffusion and mapping of the central nervous system]. PMID- 17427682 TI - [Evolution of the hospital institutions of Brussels and the progressive individualization of the departments of obstetrics]. AB - The origin of Brussels hospitals goes back to the XIIth century when several institutions created by the Church were only aimed at lodging poor pilgrims. The evolution from ecclesiastic management to municipal direction ended up in the establishment of two main public hospitals devoted to health care : Saint-Jean and Saint-Pierre. The latter, founded under Austrian rule and associated from the start with clinical teaching, gained therefore a prominent position. In 1834, it became a university hospital thanks to an agreement between the recently founded Free University of Brussels and the municipal health authorities. Finally, the administration of the main university clinical infrastructure was totally taken over by the University and moved to the newly erected Erasmus Hospital together with all faculty buildings. Development of the Brussels Maternity occurred rather slowly and underwent several location changes because of varying general politics as well as for sanitary reasons of hygiene. It was not before the XXth century that obstetrical practice progressively shifted from the private domiciliary sphere at the hands of midwives, to the hospital environment and medical control as is the case today. PMID- 17427683 TI - Laboratory evaluation of diflubenzuron as a feed-through for control of immature sand flies (Diptera: Psychodidae). AB - The benzoylurea chitin synthesis inhibitor diflubenzuron was evaluated as a rodent feed-through for the control of immature stages of Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of second instars of P. papatasi larvae that were fed feces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing 0, 8.97, 89.7, or 897 ppm diflubenzuron was evaluated. No pupation or adult emergence occurred when larvae were fed feces from hamsters that were fed diets containing diflubenzuron. The mortality of sand flies fed feces from treated hamsters was coincident with pupation of the controls, suggesting a specific effect on the larval-to-pupal molt. The results of this study suggest that a control strategy using rodent baits containing diflubenzuron for phlebotomine sand flies and zoonotic cutaneous leishmaniasis may be possible. PMID- 17427684 TI - Insecticide resistance in the bed bug: a factor in the pest's sudden resurgence? AB - Infestations of the bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), are increasing around the world at an alarming rate and have become a major public health concern. The evolution of insecticide resistance could be a primary factor in explaining this resurgence. Extremely high levels of resistance to two pyrethroid insecticides, deltamethrin and lambda-cyhalothrin, relative to a susceptible colony, were detected in populations collected from human dwellings in Kentucky and Ohio. Offspring of a cross between a resistant and susceptible colony had intermediate susceptibility. Evaluations of populations from across the United States indicate that resistance to pyrethroid insecticides is already widespread. Without the development of new tactics for bed bug management, further escalation of this public health problem should be expected. PMID- 17427685 TI - Flight muscle dimorphism and heterogeneity in flight initiation of field collected Triatoma infestans (Hemiptera: Reduviidae). AB - Recent experiments demonstrated that most field-collected Triatoma infestans (Klug) (Hemiptera: Reduviidae) adults from northern Argentina either never initiated flight or did so repeatedly in both sexes. This pattern could not be explained by sex, adult age, weight, weight-to-length ratio (W/L), or chance. We examined whether bugs that never initiated flight possessed developed flight muscles, and whether flight muscle mass relative to total body mass (FMR) was related to the probability of flight initiation. Approximately half of the adults that never initiated flight had no flight muscles. The absence of flight muscles was 2.4 times more frequent in males than females. Females had significantly larger flight muscle mass than males. For both sexes, the frequency of bugs with no flight muscles was spatially heterogeneous among individual collection sites. A logistic regression model of flight initiation that included both FMR and W/L provided abetter fit than models including either one of these predictors. FMR is a novel predictor of flight initiation in Triatominae, with a stronger effect than W/L. The higher frequency of females initiating flight in our experiments may be explained by females having flight muscles more frequently than males, and having FMR and W/L values more suitable for flying. These findings demonstrate that individuals and natural populations of T. infestans can differ dramatically with regard to flight initiation. PMID- 17427686 TI - Critical evaluation of quantitative sampling methods for Aedes aegypti (Diptera: Culicidae) immatures in water storage containers in Vietnam. AB - In response to an identified paucity of information on the size and composition of immature Aedes aegypti (L.) (Diptera: Culicidae) populations in large field containers, we assessed net sampling and pumping/sieving methods for estimating and enumerating third (III)/fourth (IV) instar and pupal populations. Sweep net detection thresholds (number above which > or = 90% chance of a positive sample) were < or = 28 immatures for seven different container types (115-3000 liter jars and tanks) in the laboratory, and mean recovery percentages varied by container type (6.15-41.29 and 7.39-33.10% for III/IV instars and pupae, respectively). A pumping method or hand bailing was applied in the field for the collection of III/IV instars and pupae from 406 receptacles, of which 343 had been previously sampled via a five-sweep netting technique. Larvae were 9.30 times more prevalent than pupae, and abundance varied by container type with means of 36-537 III/IV instars and 6-53 pupae per receptacle. Sweep netting for III/IV instars effectively identified 86.2% of Ae. aegypti-positive containers, whereas sampling for pupae detected only 43.1% of positive containers. When conversion factors (inverse of laboratory recovery percentages) were applied to field net sampling data, estimates of container populations were more accurate for III/IV instars than pupae (maximum R2 = 0.610 and 0.328, respectively); however, the relationship between immature abundance and emergent adult populations remains to be defined. PMID- 17427687 TI - Differential survival of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) larvae exposed to low temperatures in Taiwan. AB - Aedes aegypti (L.) and Aedes albopictus (Skuse) differ in their distribution in Taiwan. The former species is distributed in the south of Taiwan, whereas the latter is found throughout the island. One possible explanation proposes that low temperatures in the winter prevent the expansion of Ae. aegypti. Hence, the impact of low temperatures on immatures of both species was studied in the laboratory and in the field. Our study showed that, under most conditions, Ae. aegypti were more sensitive to low temperatures than Ae. albopictus both in the laboratory and in the field. The survival rates at 10 degrees C for the first and fourth instars of Ae. albopictus were significantly better than those of Ae. aegypti. At 2.5 and 5.0 degrees C, the first instars of Ae. albopictus survived better than the same stadium of Ae. aegypti, but the fourth instars of Ae. aegypti survived better. Short exposures to low temperatures did not affect the acclimatization of Ae. aegypti immatures but longer exposures did increase the physiological adaptation to low temperatures. For Ae. albopictus, exposure to low temperatures increases the acclimatization of this species. In field experiments, Ae. aegypti larvae had a significantly higher mortality than Ae. albopictus during exposures to cold fronts in the 2004 winter. We conclude that low temperatures in northern and central Taiwan have a negative impact on the distribution of Ae. aegypti, but this factor alone is not sufficient to prevent this species from occupying the rest of Taiwan. PMID- 17427688 TI - Early evening questing and oviposition activity by the Culex (Diptera: Culicidae) vectors of West Nile virus in northeastern North America. AB - To determine whether the Culex (Diptera: Culicidae) mosquitoes that transmit West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in the northeastern United States seek hosts and oviposit contemporaneously, we recorded when these mosquitoes attacked caged birds and when they deposited eggs. They traversed oviposition sites most frequently approximately 2 h after astronomical sunset, and eggs generally were deposited at that time. Although they most frequently approached avian hosts approximately 2 h after sunset during midsummer, they are more opportunistic during mid- to late fall. Because the Culex mosquitoes that serve as the main vectors of West Nile virus in the northeastern United States quest for hosts and seek to oviposit well after sunset, insecticidal aerosols would be most effective when applied at that time. PMID- 17427689 TI - Interspecific and intraspecific differences in foraging preferences of container dwelling mosquitoes. AB - Feeding preferences of larval container-dwelling mosquitoes are not well understood. Primary production is often absent in container systems and external inputs of animal and plant detritus supply the energy base of container food webs by supporting microorganism prey for mosquitoes. We quantified the feeding preferences of Aedes albopictus (Skuse), a non-native invasive mosquito, and Ochlerotatus triseriatus (Say), a native mosquito, when given a choice of animal and plant detritus. We tested for interpopulational (Illinois versus Florida) differences in feeding preferences, and quantified each species' performance on these two detritus types. When given a choice, both species spent significantly more time feeding at an animal detritus patch. The Illinois populations of both species spent more time feeding at animal detritus patches than did the Florida populations, which spent more time feeding at leaf detritus than did Illinois populations. Both species reached a later instar and had higher survival when reared with animal versus leaf detritus. Ae. albopictus spent more time feeding at animal detritus and had higher survival when reared on either detritus type compared with Oc. triseriatus. Greater preference for and better performance exhibited by Ae. albopictus in high-quality food (animal detritus) may result in preemption of high quality food and may contribute to the superior competitive ability of Ae. albopictus relative to Oc. triseriatus. PMID- 17427690 TI - Taxonomic and phylogenetic relationships between neotropical species of ticks from genus Amblyomma (Acari: Ixodidae) inferred from second internal transcribed spacer sequences of rDNA. AB - The accurate specific identification of ticks is essential for the study, control and prevention of tick-borne diseases. Herein, we determined ribosomal nucleotide sequences of the second internal transcribed spacer (ITS2) of 15 Neotropical hard tick species of the genus Amblyomma Koch found in Brazil. Most of the studied ticks accidentally parasite humans and potentially act as vectors of zoonoses. Lengths of the ITS2 sequences ranged from 956 to 1,207 bp, whereas GC content varied from 62.4 to 66.9%. A matrix of ITS2 divergence was calculated with the ITS2 sequence data obtained showing divergence levels varying from 1.5 to 28.8%. The analysis indicated that this molecular marker can be useful for Amblyomma specific identification. Phylogenetic inferences based on the ITS2 sequences were used to assess some issues in subgenus taxonomy. PMID- 17427691 TI - A detailed larval salivary gland polytene chromosome photomap for Culex quinquefasciatus (Diptera: Culicidae) from Johannesburg, South Africa. AB - The suitability of polytene chromosomes for population genetic and in situ DNA physical mapping was investigated in Johannesburg, South Africa, Culex quinquefasciatus Say and Culex pipiens L.. The most defined and consistent polytene spreads were obtained from salivary glands of early fourth instars from Cx. quinquefasciatus. A photomap is provided for a Cx. quinquefasciatus colony originating from Johannesburg. Variations in banding patterns, chromosome aberrations, and paracentric inversions within the colony and comparisons with previous published descriptions are discussed. Chromosomes of sufficient quality were obtained such that a DNA coding for an odorant binding protein could be mapped by colorimetric in situ hybridization. PMID- 17427692 TI - Collection and analysis of salivary proteins from the biting midge Culicoides nubeculosus (Diptera: Ceratopogonidae). AB - Salivary proteins of hematophagous Culicoides spp. are thought to play an important role in pathogen transmission and skin hypersensitivity. Analysis of these proteins, however, has been problematic due to the difficulty in obtaining adequate amounts of secreted Culicoides saliva. In the current study, a collection method for midge saliva was developed. Over a 3-d period, 3- to 5-d old male and female Culicoides nubeculosus Meigen (Diptera: Ceratopogonidae) were repeatedly placed onto the collection system and allowed to deposit saliva into a filter. Salivary products were eluted from the filters and evaluated by gel electrophoresis and mass spectrometry as well as by intradermal testing and determination of clotting time. Gel electrophoresis revealed approximately 55 protein spots displaying relative molecular masses from 5 to 67 kDa and isoelectric points ranging from 4.5 to 9.8. The majority of molecular species analyzed by mass spectrometry showed high convergence with salivary proteins recently obtained from a cDNA library of Culicoides sonorensis Wirth & Jones, including proteins involved in sugarmeal digestion, defense, and coagulation inhibition as well as members of the D7 family and unclassified salivary proteins. In addition, the proteome analysis revealed a number of peptides that were related to proteins from insect species other than Culicoides. Intradermal injection of the saliva in human skin produced edema, vasodilatation, and pruritus. The anticoagulant activity of the saliva was demonstrated by significantly prolonged clotting times for human platelets. The potential role of the identified salivary proteins in the transmission of pathogens and the induction of allergies is discussed. PMID- 17427693 TI - Biological activity of yangambin on the postembryonic development of Chrysomya megacephala (Diptera: Calliphoridae). AB - Phytochemicals endowed with hormonal, antihormonal, or toxic activity are potential agents for insect control. Thus, we became interested in testing Brazilian plant metabolites on Chrysomya megacephala (F.) (Diptera: Calliphoridae), a public health menace that is one of the most prevalent flies in Brazilian urban areas. We tested the lignan yangambin, from the leaves of Ocotea duckei Vattimo (Lauraceae). Topical treatment of eggs and first instars with yangambin as well as feeding larvae a yangambin-treated diet resulted in inhibition of postembryonic development, morphological alteration, and oviposition reduction. PMID- 17427694 TI - Impact of a bifenthrin-treated lethal ovitrap on Aedes aegypti oviposition and mortality in north Queensland, Australia. AB - Lethal ovitraps (LOs) containing an insecticide-treated ovistrip are used as a lure-and-kill device for the container-breeding dengue vector, Aedes aegypti (L.). We aimed to affirm that the pyrethroid bifenthrin could be used effectively in LOs against Ae. aegypti in north Queensland, Australia, by quantifying oviposition in and mortality caused by LOs. Small cage experiments in which individual gravid Ae. aegypti were given a choice of LOs and untreated ovitraps revealed that although LOs were less acceptable for oviposition, they provided an average 64.6% adjusted mortality. Although 92% of mosquitoes ovipositing in LOs died, 61.8% of mosquitoes that visited but did not oviposit in an LO also died, demonstrating that lethal contact occurred without egg laying. The bifenthrin content of strips (approximately 0.1 mg/cm2; 7 mg/strip) did not decrease significantly after 4 wk of field exposure nor did the toxic effect of the LOs. Large cage trials with groups of 10 Ae. aegypti confirmed that bifenthrin-treated LOs provided consistent control (average adjusted mortality 79.7%). Four-week field trials in north Queensland showed that although LOs were acceptable to ovipositing Ae. aegypti (mean time to first egg 10.9 d; mean eggs 47.3), insecticide-free ovitraps were oviposited in more readily (6.8 d, 199 eggs). The number of eggs laid per mosquito in laboratory LOs allowed calculation of the number of Ae. aegypti killed in field-deployed LOs; rapid estimates can be made by simply dividing the number of eggs on the strip by 2.84. Overall, the studies demonstrated that bifenthrin-treated LOs have potential for use as a lure-and kill device against Ae. aegypti and that they should be effective in the field for at least 4 wk. Given that untreated ovitraps were more acceptable for Ae. aegypti oviposition, the removal of alternative oviposition sites before deployment of LOs in the field should maximize their effectiveness. PMID- 17427695 TI - Structure-activity relationships of 33 piperidines as toxicants against female adults of Aedes aegypti (Diptera: Culicidae). AB - Aedes aegypti (L.) (Diptera: Culicidae) is the primary vector of both dengue and yellow fever. Use of insecticides is one of the primary ways to control this medically important insect pest. However, few new insecticides have been developed for mosquito control in recent years. As a part of our collaborative effort to search for new insecticides to control mosquitoes, piperidine was used as base compound for further optimization. Herein, we report the structure activity relationships of 33 piperidines against adult female Ae. aegypti. On the basis of 24-h LD50 values after topical application, the most toxic compound was 2-ethyl-piperidine, with an LD50 as low as 0.8 microg per mosquito. The toxicities of piperidine derivatives were significantly decreased when a benzyl moiety was attached to the carbon of the piperidine ring, with an LD50 value as high as 29.2 microg per mosquito. The toxicity order of three moieties attached to the carbon of the piperidine ring was ethyl- > methyl- > benzyl-derivatives. When the same moiety was attached to the piperidine ring, the carbon position to which the moiety was attached conferred different toxicity and the toxicity order was second carbon > third carbon > fourth carbon. Together, these preliminary results may be useful in guiding further piperidine ring modifications in the development of potential new insecticides. PMID- 17427696 TI - Evaluation of indoxacarb, an oxadiazine insecticide for the control of pyrethroid resistant Anopheles gambiae (Diptera: Culicidae). AB - Owing to the spread of pyrethroid resistance in Anopheles gambiae s.s. (Diptera: Culicidae) and other vector mosquitoes, there is an urgent need to develop alternative insecticides to supplement the pyrethroids for malaria control. Indoxacarb is an oxadiazine insecticide initially commercialized by DuPont for control of agricultural pests. Performance against An. gambiae bearing kdr (pyrethroid and DDT resistance) or Ace-1R insensitive acetylcholinesterase (organophosphate and carbamate resistance) mechanisms was studied using larval and adult bioassays and a simulated experimental hut system (tunnel tests) that allows fuller expression of the behavioral responses to insecticide. Larval and adult bioassays (topical application and cone tests on treated netting) showed a standard probit dosage-mortality response and no evidence of cross-resistance to the kdr and Ace-1R resistance mechanisms. Toxic activity was slow compared with standard insecticides and additional mortality was observed. Indoxacarb induced no excitorepellency in adults. In tunnel tests, indoxacarb induced no inhibition of mosquito penetration or blood feeding through the holed netting, but it induced delayed mortality over 24-96 h. There was > 90% mortality of the kdr strain on netting treated with the 500 mg/m2 dosage, whereas permethin at 500 mg/nm2 only killed 30% of this strain. A mixture of indoxacarb and pyrethroid showed neither synergism nor antagonism. The absence of cross-resistance to current insecticides indicates that indoxacarb has malaria vector control potential as larvicide or adulticide where mosquitoes are pyrethroid resistant. PMID- 17427697 TI - Efficacy of a single doramectin injection against adult female Boophilus microplus (Acari: Ixodidae) in the final stages of engorgement before detachment. AB - The efficacy of injectable doramectin applied at 200 microg/kg was evaluated against adult female Boophilus microplus (Canestrini) (Acari: Ixodidae) in the later stages of engorgement before detachment. Lethal levels of doramectin in the serum of treated cattle (9.7-36.6 ppb) were reached and sustained within 24 h after treatment. However, treatment at 19 or 20 d postinfestation allowed greater tick survival and reproductive capability (IF) than at 18 d postinfestation, indicating that a significant portion of the ticks were able to engorge and detach before obtaining a lethal dose of doramectin. Thus, treatment at 18 d after infestation provided significantly higher overall control (99.5%) than treatment at 19 or 20 d postinfestation (95.8 and 89.1%, respectively). Analysis of control on a daily basis demonstrated that treatment at 18 d postinfestation provided > 99% on each day of the evaluation. Conversely, treatment at 19 or 20 d postinfestation produced levels of control ranging from 22.6 to 85.6% during the first 2 d of female detachment, and > or = 99% control was not achieved until after the fourth day of female detachment, where it remained throughout the study. Therefore, application of injectable doramectin at < or = 18 d after tick infestation was the only treatment regime considered acceptable for use in the U.S. Boophilus Eradication Program. Treatment intervals > 18 d postinfestation could pose a substantial risk of dispersing viable ticks to tick-free areas outside the permanent quarantine zone that has been established along the Texas Mexico border. PMID- 17427698 TI - A comparison of three bioassay techniques to determine amitraz susceptibility in Boophilus microplus (Acari: Ixodidae). AB - The ability of the Miller, Soberanes, and White bioassay techniques to describe amitraz susceptibility in three different strains of Boophilus microplus (Canestrini) (Acari: Ixodidae) was compared. For a susceptible strain, all techniques adequately described amitraz susceptibility by producing a full range of mortality that corresponded with increasing concentration of amitraz. However, when resistant strains were evaluated, only the Miller and the Soberanes techniques adequately estimated the dose-response relationship. Lethal concentrations were not precisely estimated when all the data were included in the analyses for every strain and technique tested. Better estimates were obtained when subsets of data around the range of interest were subjected to probit analysis. For the Soberanes technique, the slope of the probit regression was steeper for the Brazilian resistant and Texan susceptible strains compared with the heterozygous Mexican strain. The pattern was different when the same strains were tested with the Miller technique. The slopes of the regressions for the Mexican and the Texan strains did not differ significantly, but the Brazilian strain had a steeper slope than the other strains. Resistance ratios were much greater when the Soberanes technique was used than when the Miller technique was used on the same strains. However, neither technique produced enough separation between susceptible and resistant strains to develop a traditional discriminating dose (DD) test that required a concentration of 2x LC99.9 estimate. A DD test at the LC99 would be possible for both techniques. We discuss the strengths and weaknesses of the three techniques, including potential improvements to the White technique. The White technique has the greatest potential to determine the mechanisms of amitraz resistance in detailed synergist studies. Currently, only the Miller method can fulfill this task. The Miller and Soberanes techniques are well suited for the study of the epidemiology of resistance worldwide, because they use commercially available, formulated amitraz that is easy and inexpensive to obtain. PMID- 17427699 TI - Response of Armigeres subalbatus (Diptera: Culicidae) to intraperitoneally isolated Brugia spp. microfilariae. AB - The relationship between mosquito and parasite involves a delicate balance that is influenced not only by the mosquito but also by parasite determinants. Using the biologically and morphologically similar parasites Brugia malayi and Brugia pahangi and the mosquito Armigeres subalbatus (Coquillett) (Diptera: Culicidae), it should be possible to dissect out the key elements involved in initiating or avoiding an immune response, known as melanotic encapsulation, because in this mosquito B. malayi microfilariae (mf) are melanized and destroyed, but B. pahangi mf develop normally into infective-stage larvae. Because of limitations in isolating sufficient mf from the circulation of an infected mammalian host, Brugia spp. mf that can be obtained in large numbers from the peritoneal cavity of an infected host were tested to ascertain the immune response of Ar. subalbatus to this source of mf. Results indicate that the immune response of Ar. subalbatus against intraperitoneal (i.p.) Brugia spp. mf mimics that which is observed when this mosquito is exposed to mf-infected animals, indicating that i.p. mf are similar to those mf that circulate naturally in the blood of the vertebrate host. Therefore, the i.p. mf should serve as an excellent source of material for genomic and proteomic studies designed to analyze the role of the parasite in influencing the immune response of the mosquito. PMID- 17427700 TI - Is nonviremic transmission of West Nile virus by Culex mosquitoes (Diptera: Culicidae) nonviremic? AB - Inter- and intraspecific transfer of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) occurred infrequently when donor Culex tarsalis Coquillett fed concurrently on house finches with recipient Culex quinquefasciatus Say and Cx. tarsalis. Five of six of these house finches had WNV in blood samples collected by jugular venipuncture 30-45 min postfeeding, with titers ranging from 2.3 to 4.2 log10 plaque-forming units (PFU)/ml. After 2-wk incubation at 26 degrees C, three Cx. quinquefasciatus and one Cx. tarsalis of 230 blood-fed recipients were infected, of which one Cx. quinquefasciatus was capable of transmission. Our data indicated that infectious female mosquitoes feeding on small vertebrates create a nonpropagative viremia capable of infecting concurrently cofeeding females. The proportion of recipients infected is likely related to the amount of virus expectorated by donor females, the blood volume of the vertebrate host, and the susceptibility of the cofeeding mosquitoes. PMID- 17427701 TI - Borrelia lusitaniae in immature Ixodes ricinus (Acari: Ixodidae) feeding on common wall lizards in Tuscany, central Italy. AB - Lizards and small rodents were live captured in Tuscany, central Italy, from May through August 2005. Prevalence of infestation by larval Ixodes ricinus L. (Acari: Ixodidae) and mean numbers of larvae per host were not significantly different for common wall lizards, Podarcis muralis Laurenti, and Apodemus spp. mice, whereas infestation levels by nymphs were significantly greater on lizards. Borrelia lusitaniae, which was previously shown to be dominant in host-seeking I. ricinus in the same study area, was detected by polymerase chain reaction (PCR) in 19.8% (95% confidence interval: 14.4, 26.0) of larval ticks and in 52.9% (27.8, 77.0) of nymphs that were collected from lizards. Moreover, 18.8% (7.2, 36.4) and 25.0 (3.2, 65.1) of lizards' tail biopsies and blood samples, respectively, were positive for B. lusitaniae. Conversely, attached ticks and ear biopsies from Apodemus spp. mice were PCR negative. Passerine birds belonging to 10 species were live captured in March 2005, and Borrelia valaisiana was detected in 57.1% (18.4, 90.1) of I. ricinus nymphs feeding on Eurasian blackbirds, Turdus merula L. Results of this study suggest that lizards play an important role as reservoirs for B. lusitanae and may affect the dominance of this genospecies in the Mediterranean area. PMID- 17427702 TI - First report of permethrin-resistant Boophilus microplus (Acari: Ixodidae) collected within the United States. AB - Boophilus microplus (Canestrini) (Acari: Ixodidae), collected in Hidalgo County, TX, were determined to be resistant to permethrin. Discriminating dose (DD) tests at the LC99 and 2X the LC99 of susceptible ticks yielded lower than expected mortalities for permethrin but not for coumaphos or amitraz acaricides. Initial bioassay results confirmed the pyrethroid resistance detected in the DD assays. Two generations of selection with permethrin at a rate > 60% increased the measured resistance ratios from 9.5 (7.9-11.5) to 263 (217-320). Synergist studies suggested that metabolic enzymes were involved in permethrin resistance. Native gel electrophoresis verified that the CZS9 esterase was not involved in resistance to permethrin. Polymerase chain reaction examination revealed the presence of a mutation of the sodium channel gene (Phe [arrow] Ile amino acid substitution in the S6 trans-membrane segment of domain III), in the resistant B&H population. The frequency of this mutation increased after selection with permethrin and concurrent increase in estimated resistance ratios. The B&H population was eradicated from the United States by the U.S. Department of Agriculture's Animal and Plant Health Inspection Service Veterinary Services, Cattle Fever Tick Eradication Program through the use of the organophosphate acaricide coumaphos. PMID- 17427703 TI - Does feeding on infected mosquitoes (Diptera: Culicidae) enhance the role of song sparrows in the transmission of arboviruses in California? AB - Song sparrows, Melopiza melodia, inoculated subcutaneously with either western equine encephalomyelitis virus (family Togaviridae, genus Alphavirus, WEEV) or West Nile virus (family Flaviviridae, genus Flavivirus, WNV) developed elevated viremias, and they were considered to be competent experimental hosts for both viruses. However, birds that ingested from three to 20 mosquitoes containing comparable amounts of either WEEV or WNV failed to become infected, indicating limited oral susceptibility. Comparatively few field-collected birds had antibodies against either WEEV or WNV, indicating that this species was infrequently bitten by infectious mosquitoes in nature and probably was of limited importance in viral amplification. PMID- 17427704 TI - Ability of transstadially infected Ixodes pacificus (Acari: Ixodidae) to transmit West Nile virus to song sparrows or western fence lizards. AB - The hypothesis that Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) may serve as a reservoir and vector of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in California was tested by determining the ability of this tick species to become infected with the NY99 strain of WNV while feeding on viremic song sparrows, to maintain the infection transstadially, and then to transmit WNV to recipient naive song sparrows and western fence lizards during the nymphal stage. The percentage of ticks testing positive by reverse transcription polymerase chain reaction (RT-PCR) decreased from 77% of 35 larvae at day 6 after ticks were transferred to donor song sparrows (day of detachment) to 23% of 35 nymphs at 59 d postinfestation (approximately 19 d after molting to the nymphal stage). However, the percentage of ticks positive by RT-PCR from which infectious virus was recovered by Vero cell assay decreased from 59% on day 6 to 12% on day 59, even though there was no statistically significant decrease in the quantity of RNA within positive ticks. Attempts to improve the sensitivity of plaque assays by blind passage through C6/36 cell cultures were unsuccessful. These data indicated that ticks maintained viral RNA but not necessarily infectious virus over time. Nymphs from larvae that fed on song sparrows with peak viremias ranging from 7.2 to 8.5 log10 plaque-forming units (PFU) per ml were used in transmission attempts. From one to seven RNA-positive nymphal ticks engorged and detached from each of four recipient song sparrows or western fence lizards. Blood samples from sparrows and lizards remained negative, indicating that transmission did not occur. An additional four lizards inoculated with 1,500 PFU of WNV developed moderate viremias, ranging from 4.2 to 5.6 log10 PFU/ml. Our data and data from previous studies collectively indicated that ixodid ticks were not able to experimentally transmit WNV and therefore most likely would not be important vectors in WNV transmission cycles. PMID- 17427705 TI - Ectoparasite diversity and exposure to vector-borne disease agents in wild rodents in central coastal California. AB - A survey of wild rodents was performed in the Morro Bay area of central coastal California to determine serological and polymerase chain reaction (PCR) prevalence of Anaplasma phagocytophilum Dumler, Barbet, Bekker, Dasch, Palmer, Ray, Rikihisa, and Rurangirwa, Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt, and Brenner, Francisella tularensis McCoy, and Yersinia pestis Yersin; to describe the ectoparasitic fauna on important vector-borne disease hosts; and to determine whether pathogen exposure was associated with infestation by ectoparasites. We trapped 411 rodents from 10 species in 2004 and 2005. Anaplasma phagocytophilum exposure was detected in 11% of all wild rodents tested, with seropositive animals in eight species. Anaplasma phagocytophilum DNA was detected by PCR amplification in Neotoma fuscipes Baird and Reithrodontomys megalotis Baird (0.6% of all rodents). Yersinia spp. exposure was identified in 3.2% of all rodents tested, with highest detected exposure in peridomestic rodents, Mus musculus L. (20%), and Rattus rattus L. (50%). No individuals tested positive for the Y. pestis pla gene by PCR. In total, 338 fleas were identified from each of 10 rodent species examined. The most abundant flea was Malareus telchinus Rothschild. Relative density of flea infestation was highest on Spermophilus beecheyi Richardson and Microtus californicus Peale. Ticks recovered from trapped animals included Ixodes angustus Neumann, Ixodes pacificus Cooley & Kohls, Ixodes spinipalpis Hadwen & Nuttall, and Dermacentor occidentalis Marx. Given the moderate climate and diversity of rodents and arthropods in the Morro Bay area, ongoing investigation of this region will be helpful in understanding disease maintenance cycles. PMID- 17427706 TI - Mosquito species succession and physicochemical factors affecting their abundance in rice fields in Mwea, Kenya. AB - The succession of mosquito species and abiotic factors affecting their distribution and abundance in rice (Oryza spp.) fields was investigated over a 16 wk rice growing cycle covering the period between January and May 2006. Fifteen experimental rice plots were sampled for mosquito larvae and characterized based on rice height, number of tillers, floating vegetation cover, water depth, water temperature, turbidity, salinity, pH, dissolved oxygen, total dissolved solids, and conductivity. Microscopic identification of 3,025 larvae yielded nine mosquito species predominated by Anopheles arabiensis Patton (45.0%), Culex quinquefasciatus Say (35.8%), Anopheles pharoensis Theobald (9.0%) and Ficalbia splendens Theobald (7.1%). Other species, including Anopheles rufipes Gough, Anopheles coustani Laveran, Anonopheles maculipalpis Giles, Culex annulioris Theobald, and Culex poicilipes Theobald made up 3.1% of the total collection. Anopheles gambiae s.l., Cx. quinquefasciatus, and An. pharoensis occurred throughout the cycle, but they were more abundant up to 4 wk posttransplanting with peaks after fertilizer application. As rice plants became established, three groups of mosquitoes were recognized: the first groups included An. rufipes, Fl. splendens, and Cx. annulioris, which occurred throughout much of the second half of the rice cycle, whereas the second group included Cx. poicilipes, which was found in the middle of the rice cycle. An. coustani and An. maculipalpis formed the third group occurring toward the end of the cycle. Dissolved oxygen, number of tillers, and rice height were negatively associated with the abundance ofAn. arabiensis and Cx. quinquefasciatus larvae. In addition, Cx. quinquefasciatus also was associated with water depth (-ve) and turbidity (+ve). Abundance of An. pharoensis larvae was significantly associated with water temperature (+ve), the number of tillers (-ve), and rice height (-ve), whereas Fl. splendens was significantly associated with the number of tillers (+ve). The results demonstrate a complex nature of the interactions between some of the factors in the ecosystem and mosquito species abundance and calls for time-dependent and species-specific mosquito control operations. PMID- 17427707 TI - Aedes aegypti population sampling using BG-Sentinel traps in north Queensland Australia: statistical considerations for trap deployment and sampling strategy. AB - BG-Sentinel mosquito traps were trialed as a tool for the rapid assessment (24-h collections) and routine monitoring (72-h collections) of adult Aedes aegypti L. populations in north Queensland. Analysis of Ae. aegypti collections using BG Sentinels set in suburban Cairns for 24 h permitted the calculation of sample size for a range of precision levels. Clusters of houses with BG-Sentinels operating continuously for 15 d, with collections every 72 h, also permitted required sample size calculation. Evidence of Ae. aegypti spatial clustering at the house scale was revealed, with statistically significant effects detected for all collection days. Less variation was detected at each trap location, with only nine of 32 trap locations revealing significant clustering over time. Trap-out effects through continuous BG-Sentinel operation at a fixed location were absent. The findings support fixed position sampling at 72-h intervals for routine monitoring ofAe. aegypti populations in Cairns. Despite the relationship between collections of adult vectors and the incidence of disease remaining unknown, BG Sentinel collections provide an alternative and less labor-intensive abundance measure for assessing risk of dengue virus transmission and success of dengue vector control programs. PMID- 17427708 TI - Light-induced variability in development of forensically important blow fly Phormia regina (Diptera: Calliphoridae). AB - The use of the postmortem interval (PMI) in practical applications of forensic entomology is based upon developmental data of blow flies (Diptera: Calliphoridae) generated under controlled environmental conditions. Careful review of the published forensic entomology data sets showed that experimental (environmental) parameters differed between studies. Despite the differences in study design, there are no empirical data on the effect of photoperiod on blow fly development; yet, photoperiod has been shown to alter some insect development and behavior among noncalliphorids. Consequently, will differences in photoperiod alter the developmental times of calliphorids, and thereby alter PMI estimates? To answer this question, we used a replicated design with precise temperature measurement to examine the effects of photoperiod on the forensically important blow fly Phormia regina (Meigen). We concluded that inaccurate temperature recordings by using set-chamber temperatures over rearing-container temperatures would have overshadowed any affect light had on development. Second, constant light increased variation in overall adult developmental time and significantly delayed development compared with cyclic light. Finally, not accounting for delayed development induced by photoperiod underestimated the initial empirical estimate of the PMI. These sources of variation need to be included in forensic estimates because this variation can compromise predictions of PMI based upon current data sets. Without pinpointing optimal photoperiods with which to test development, we must assume that potentially large sources of variability exist within current estimates of the PMI. PMID- 17427709 TI - Seasonal pattern of host-seeking activity by the human-biting adult life stage of Dermacentor andersoni (Acari: Ixodidae). AB - Nonremoval drag sampling was conducted in multiple sites in Larimer County, CO, from March to July 2006 to determine the seasonal pattern of host-seeking activity by the human-biting adult life stage of the Rocky Mountain wood tick, Dermacentor andersoni Stiles (Acari: Ixodidae). Four sites, at elevations ranging from 1,790 to 2,470 m, consistently yielded host-seeking ticks. Ticks were active from March until late June; the length of the active period ranged from 84 to 104 d. Based on site-specific linear regression models of increase and decrease in tick host-seeking activity over time, densities of D. andersoni adults were estimated to reach 50% of their peaks between 2 and 11 April, to peak between 21 and 28 April, and to fall below 50% of the peaks between 24 and 27 May. The length of the periods with tick densities exceeding 50 and 75% of the peak was 43 52 and 21-26 d, respectively. Rapid increases in tick numbers in late March to early April followed a stretch of days with daily maximum temperatures exceeding 5 degrees C, whereas rapidly declining tick numbers in mid- to late May were associated with daily maximum temperatures consistently exceeding 20 degrees C and daily minimum relative humidity commonly falling below 20%. Densities of D. andersoni adults exceeded 50% of the peak when daily maximum temperatures were in the 16-19 degrees C range and daily minimum relative humidity was > 20%. Finally, tick seasonality may be adapted to local climatic conditions within Larimer County; site-specific daily maximum temperatures at the time of peak tick host seeking activity in late April were positively associated with site-specific mean daily maximum temperatures for April. PMID- 17427710 TI - Olfactory responses of medically and economically important mites (Acari: Epidermoptidae and Acaridae) to volatile chemicals. AB - Dermatophagoides farinae Hughes (Acari: Epidermoptidae), the American house dust mite, and Tyrophagus putrescentiae (Schrank) (Acari: Acaridae), the mold mite, are medically and economically important but controlling them has proved difficult, and recolonization is commonplace. Their behavioral responses to different sources of volatile chemicals are still not fully elucidated. For the first time, the Y-tube olfactometer, which is an enclosed bioassay to resolve responses to test and control volatiles, has been successfully used with these mites. Mites were tested individually, and both T. putrescentiae and D. farinae responded to food volatiles. Y-tube olfactometers may be used to test for potential semiochemicals, thereby increasing knowledge of our behavior of astigmatic mites. PMID- 17427711 TI - Duration of exposure to suboptimal atmospheric moisture affects nymphal blacklegged tick survival. AB - The biological processes affecting Ixodes scapularis Say survival are complex. Understanding these processes will be beneficial for predicting tick distribution and population dynamics. This research shows that the duration for which nymphal ticks are exposed to drying air is an important factor for their survival. Experimental analysis of variance results show that duration of exposure to dry air (duration) is as important as vapor pressure deficit (relative humidity) (duration, relative humidity, P < 0.0001). Ticks do not survive when exposed to dry air for long periods; however, the return of humid air within 4-8 h has as large a positive impact on tick survival, as does constant humid air. This experiment exposes nymphal ticks to conditions of suboptimal humidity for different durations and then returns them to saturated conditions that are more typical of daily relative humidity fluctuations experienced during summer in southern New England forests. PMID- 17427712 TI - A polymerase chain reaction-based diagnostic to identify larvae and eggs of container mosquito species from the Australian region. AB - Dengue outbreaks occur regularly in parts of northern Queensland, Australia, and there is concern that these outbreaks may spread with the introduction and range expansion of the two main vectors Aedes aegypti (L.) and Aedes albopictus (Skuse). Problems encountered in separating larvae of endemic and exotic container mosquito species resulted in the development of a polymerase chain reaction diagnostic procedure that uses a restriction enzyme to cut the internal transcribed spacer region 1 of the ribosomal DNA to separate Ae. aegypti and Ae. albopictus from a number of common local container mosquito species which can be used at any stage of the life cycle, including eggs up to 8 wk of age. Identification was possible using desiccated or alcohol-preserved specimens with a response time of < 24 h after receipt of the specimens. PMID- 17427713 TI - Impact of phlebotomine sand flies on U.S. military operations at Tallil Air Base, Iraq: 3. Evaluation of surveillance devices for the collection of adult sand flies. AB - We evaluated the effectiveness of commercially available light traps and sticky traps baited with chemical light sticks for the collection of phlebotomine sand flies. Evaluations were conducted at Tallil Air Base, Iraq, in 2003. In an initial study, a Centers for Disease Control and Prevention (CDC)-style trap with UV bulb collected significantly more sand flies than did an up-draft CDC trap, a standard down-draft CDC trap (STD-CDC), or a sticky strap with a green chemical light stick. In a subsequent study, we found that the addition of chemical light sticks to sticky traps resulted in a significant increase in the number of sand flies collected compared with sticky traps without the light sticks. These data indicate that 1) the CDC light trap with an UV bulb is an effective alternative to the standard CDC light trap for collecting phlebotomine sand flies in Iraq, and 2) that the addition of a chemical light stick to a sticky trap can result in a field-expedient tool for the collection of sand flies. PMID- 17427714 TI - Phenology of Phlebotomus papatasi (Diptera: Psychodidae) relative to the seasonal prevalence of zoonotic cutaneous leishmaniasis in central Tunisia. AB - The population density of Phlebotomus papatasi Scopoli (Diptera: Psychodidae), vector of Leishmania major Yakimoff & Schokhor (Kinetoplastida: Trypanosomatidae), the etiologic agent of zoonotic cutaneous leishmaniasis (ZCL), was assessed May-November 2005 in central Tunisia by using sticky traps. The densities of P. papatasi were found to peak in early spring and again in the autumn. The lowest densities were observed in August. Prevalence of ZCL in the governorate of Sidi Bouzid peaks in December, 3 to 3.5 mo after the fall sand fly population peak, suggesting a close temporal association with the abundance of P. papatasi. PMID- 17427723 TI - A possible solution to South Africa's healthcare challenges? PMID- 17427724 TI - Socio-economic inequities in dental caries experience of 12-year-old South Africans: policy implications for prevention. AB - OBJECTIVES: To examine the socioeconomic inequities in dental caries experience of 12-year-olds, in order to inform policy actions for caries prevention in South Africa. METHODS: Secondary analysis of the 1999-2002 national survey data of 12 year-olds (N=5411), available from 37 regions in 7 of the 9 provinces, was carried out. Logistic regression was used to determine risks for caries experience among 12-year-olds in each province, using parental occupation and racial group as independent variables. Regression curve-estimation was used to examine the spatial relationship between mean DMFT and caries prevalence. RESULTS: The mean DMFT (+/-SD) for the study population was 1.19 (+/-2.13), significant caries index was 3.35 and caries prevalence was 40.1%. The highest mean DMFT was among the Coloured population (2.14+/-2.50). Compared to children in the highest occupational class, the risk for children of the unemployed to experience caries was lower in the North West province (Odds ratio [OR]=0.47; p<0.01), but significantly higher in the coastal provinces-- KwaZulu-Natal and Northern Cape, with OR of 1.32 and 1.52 respectively. The regression curve derived demonstrated that a unit increase in caries experience in low-level caries populations would generate more cases than similar increase in high-level caries populations. CONCLUSIONS: DMFT alone provided an incomplete picture of the impact of caries in South Africa, thus the need to monitor inequities as part of policy impact. The distribution of caries suggests that 'high-risk' approach to prevention in the presence of existing social gaps may inadvertently reinforce inequities in caries-burden and supports the concurrent implementation of population-approach, such as water fluoridation. PMID- 17427725 TI - Managing the polymerization shrinkage of resin composite restorations: a review. AB - Polymerization shrinkage (PS) of resin composites almost invariably generates a degree of stress at the tooth/restoration interface. The magnitude of the stresses so produced depends on resin composite composition and its ability to flow before solidification, which is, in turn related to cavity configuration and curing characteristics of the composite. The purpose of the present article is to review the published literature on the management of PS, with special reference to those elements that are under the control of the dentist. Available data from laboratory, finite element analysis and review articles, relating to the methods and techniques that have been proposed for managing the problem of PS, are included. On the basis of the review, it may be concluded that PS has negative effects on the resin composite/tooth interface. The review identifies the preferred handling and/or management techniques that clinicians might employ to reduce the levels of PS that might affect the restorations they place, and consequently reduce its adverse effects. PMID- 17427726 TI - Implant failure due to titanium hypersensitivity/allergy?--Report of a case. AB - Six titanium implants were placed in the mandible between the left and right mental foramen of a 49-year old female patient. The implants were all constructed of the same grade IV titanium. Three types of implants were used, namely LIBB compression implant, cylindrical implant and Branemark-like implant. These three types of implants were placed as part of a research project, which received approval from the ethics committee. The patient developed a severe reaction to all the implant units, with both clinical and radiological features of complication. The localised tissue reaction was severe enough to warrant removal of all the implants. The surrounding soft tissue was submitted for histological analysis, which revealed a chronic inflammatory response with concomitant fibrosis around all the implants as well as foreign body giant cell reaction around two implants. Following implant removal the patient recuperated well and the soft and hard tissue healed satisfactorily. PMID- 17427728 TI - Sedation corner 15. PMID- 17427729 TI - General practitioner's radiology case 48. Mucous retention cyst. PMID- 17427730 TI - Protect your income. PMID- 17427731 TI - Brain functional localization: a survey of image registration techniques. AB - Functional localization is a concept which involves the application of a sequence of geometrical and statistical image processing operations in order to define the location of brain activity or to produce functional/parametric maps with respect to the brain structure or anatomy. Considering that functional brain images do not normally convey detailed structural information and, thus, do not present an anatomically specific localization of functional activity, various image registration techniques are introduced in the literature for the purpose of mapping functional activity into an anatomical image or a brain atlas. The problems addressed by these techniques differ depending on the application and the type of analysis, i.e., single-subject versus group analysis. Functional to anatomical brain image registration is the core part of functional localization in most applications and is accompanied by intersubject and subject-to-atlas registration for group analysis studies. Cortical surface registration and automatic brain labeling are some of the other tools towards establishing a fully automatic functional localization procedure. While several previous survey papers have reviewed and classified general-purpose medical image registration techniques, this paper provides an overview of brain functional localization along with a survey and classification of the image registration techniques related to this problem. PMID- 17427732 TI - A deformable registration method for automated morphometry of MRI brain images in neuropsychiatric research. AB - Image registration methods play a crucial role in computational neuroanatomy. This paper mainly contributes to the field of image registration with the use of nonlinear spatial transformations. Particularly, problems connected to matching magnetic resonance imaging (MRI) brain image data obtained from various subjects and with various imaging conditions are solved here. Registration is driven by local forces derived from multimodal point similarity measures which are estimated with the use of joint intensity histogram and tissue probability maps. A spatial deformation model imitating principles of continuum mechanics is used. Five similarity measures are tested in an experiment with image data obtained from the Simulated Brain Database and a quantitative evaluation of the algorithm is presented. Results of application of the method in automated spatial detection of anatomical abnormalities in first-episode schizophrenia are presented. PMID- 17427733 TI - Large deformation diffeomorphism and momentum based hippocampal shape discrimination in dementia of the Alzheimer type. AB - In large-deformation diffeomorphic metric mapping (LDDMM), the diffeomorphic matching of images are modeled as evolution in time, or a flow, of an associated smooth velocity vector field v controlling the evolution. The initial momentum parameterizes the whole geodesic and encodes the shape and form of the target image. Thus, methods such as principal component analysis (PCA) of the initial momentum leads to analysis of anatomical shape and form in target images without being restricted to small-deformation assumption in the analysis of linear displacements. We apply this approach to a study of dementia of the Alzheimer type (DAT). The left hippocampus in the DAT group shows significant shape abnormality while the right hippocampus shows similar pattern of abnormality. Further, PCA of the initial momentum leads to correct classification of 12 out of 18 DAT subjects and 22 out of 26 control subjects. PMID- 17427734 TI - Simultaneous registration and parcellation of bilateral hippocampal surface pairs for local asymmetry quantification. AB - In clinical applications where structural asymmetries between homologous shapes have been correlated with pathology, the questions of definition and quantification of "asymmetry" arise naturally. When not only the degree but the position of deformity is thought relevant, asymmetry localization must also be addressed. Asymmetries between paired shapes have already been formulated in terms of (nonrigid) diffeomorphisms between the shapes. For the infinity of such maps possible for a given pair, we define optimality as the minimization of deviation from isometry under the constraint of piecewise deformation homogeneity. We propose a novel variational formulation for segmenting asymmetric regions from surface pairs based on the minimization of a functional of both the deformation map and the segmentation boundary, which defines the regions within which the homogeneity constraint is to be enforced. The functional minimization is achieved via a quasi-simultaneous evolution of the map and the segmenting curve, conducted on and between two-dimensional surface parametric domains. We present examples using both synthetic data and pairs of left and right hippocampal structures and demonstrate the relevance of the extracted features through a clinical epilepsy classification analysis. PMID- 17427735 TI - Atlas renormalization for improved brain MR image segmentation across scanner platforms. AB - Atlas-based approaches have demonstrated the ability to automatically identify detailed brain structures from 3-D magnetic resonance (MR) brain images. Unfortunately, the accuracy of this type of method often degrades when processing data acquired on a different scanner platform or pulse sequence than the data used for the atlas training. In this paper, we improve the performance of an atlas-based whole brain segmentation method by introducing an intensity renormalization procedure that automatically adjusts the prior atlas intensity model to new input data. Validation using manually labeled test datasets has shown that the new procedure improves the segmentation accuracy (as measured by the Dice coefficient) by 10% or more for several structures including hippocampus, amygdala, caudate, and pallidum. The results verify that this new procedure reduces the sensitivity of the whole brain segmentation method to changes in scanner platforms and improves its accuracy and robustness, which can thus facilitate multicenter or multisite neuroanatomical imaging studies. PMID- 17427736 TI - Topology-preserving tissue classification of magnetic resonance brain images. AB - This paper presents a new framework for multiple object segmentation in medical images that respects the topological properties and relationships of structures as given by a template. The technique, known as topology-preserving, anatomy driven segmentation (TOADS), combines advantages of statistical tissue classification, topology-preserving fast marching methods, and image registration to enforce object-level relationships with little constraint over the geometry. When applied to the problem of brain segmentation, it directly provides a cortical surface with spherical topology while segmenting the main cerebral structures. Validation on simulated and real images characterises the performance of the algorithm with regard to noise, inhomogeneities, and anatomical variations. PMID- 17427737 TI - A statistical parts-based model of anatomical variability. AB - In this paper, we present a statistical parts-based model (PBM) of appearance, applied to the problem of modeling intersubject anatomical variability in magnetic resonance (MR) brain images. In contrast to global image models such as the active appearance model (AAM), the PBM consists of a collection of localized image regions, referred to as parts, whose appearance, geometry and occurrence frequency are quantified statistically. The parts-based approach explicitly addresses the case where one-to-one correspondence does not exist between all subjects in a population due to anatomical differences, as model parts are not required to appear in all subjects. The model is constructed through a fully automatic machine learning algorithm, identifying image patterns that appear with statistical regularity in a large collection of subject images. Parts are represented by generic scale-invariant features, and the model can, therefore, be applied to a wide variety of image domains. Experimentation based on 2-D MR slices shows that a PBM learned from a set of 102 subjects can be robustly fit to 50 new subjects with accuracy comparable to 3 human raters. Additionally, it is shown that unlike global models such as the AAM, PBM fitting is stable in the presence of unexpected, local perturbation. PMID- 17427738 TI - Automatic segmentation of the caudate nucleus from human brain MR images. AB - We describe a knowledge-driven algorithm to automatically delineate the caudate nucleus (CN) region of the human brain from a magnetic resonance (MR) image. Since the lateral ventricles (LVs) are good landmarks for positioning the CN, the algorithm first extracts the LVs, and automatically localizes the CN from this information guided by anatomic knowledge of the structure. The face validity of the algorithm was tested with 55 high-resolution T1-weighted magnetic resonance imaging (MRI) datasets, and segmentation results were overlaid onto the original image data for visual inspection. We further evaluated the algorithm by comparing automated segmentation results to a "gold standard" established by human experts for these 55 MR datasets. Quantitative comparison showed a high intraclass correlation between the algorithm and expert as well as high spatial overlap between the regions-of-interest (ROIs) generated from the two methods. The mean spatial overlap +/- standard deviation (defined by the intersection of the 2 ROIs divided by the union of the 2 ROIs) was equal to 0.873 +/- 0.0234. The algorithm has been incorporated into a public domain software program written in Java and, thus, has the potential to be of broad benefit to neuroimaging investigators interested in basal ganglia anatomy and function. PMID- 17427739 TI - Geometrically accurate topology-correction of cortical surfaces using nonseparating loops. AB - In this paper, we focus on the retrospective topology correction of surfaces. We propose a technique to accurately correct the spherical topology of cortical surfaces. Specifically, we construct a mapping from the original surface onto the sphere to detect topological defects as minimal nonhomeomorphic regions. The topology of each defect is then corrected by opening and sealing the surface along a set of nonseparating loops that are selected in a Bayesian framework. The proposed method is a wholly self-contained topology correction algorithm, which determines geometrically accurate, topologically correct solutions based on the magnetic resonance imaging (MRI) intensity profile and the expected local curvature. Applied to real data, our method provides topological corrections similar to those made by a trained operator. PMID- 17427740 TI - A geometric method for automatic extraction of sulcal fundi. AB - Sulcal fundi are 3-D curves that lie in the depths of the cerebral cortex and, in addition to their intrinsic value in brain research, are often used as landmarks for downstream computations in brain imaging. In this paper, we present a geometric algorithm that automatically extracts the sulcal fundi from magnetic resonance images and represents them as spline curves lying on the extracted triangular mesh representing the cortical surface. The input to our algorithm is a triangular mesh representation of an extracted cortical surface as computed by one of several available software packages for performing automated and semi automated cortical surface extraction. Given this input we first compute a geometric depth measure for each triangle on the cortical surface mesh, and based on this information we extract sulcal regions by checking for connected regions exceeding a depth threshold. We then identify endpoints of each region and delineate the fundus by thinning the connected region while keeping the endpoints fixed. The curves, thus, defined are regularized using weighted splines on the surface mesh to yield high-quality representations of the sulcal fundi. We present the geometric framework and validate it with real data from human brains. Comparisons with expert-labeled sulcal fundi are part of this validation process. PMID- 17427741 TI - Automated extraction of the cortical sulci based on a supervised learning approach. AB - It is important to detect and extract the major cortical sulci from brain images, but manually annotating these sulci is a time-consuming task and requires the labeler to follow complex protocols. This paper proposes a learning-based algorithm for automated extraction of the major cortical sulci from magnetic resonance imaging (MRI) volumes and cortical surfaces. Unlike alternative methods for detecting the major cortical sulci, which use a small number of predefined rules based on properties of the cortical surface such as the mean curvature, our approach learns a discriminative model using the probabilistic boosting tree algorithm (PBT). PBT is a supervised learning approach which selects and combines hundreds of features at different scales, such as curvatures, gradients and shape index. Our method can be applied to either MRI volumes or cortical surfaces. It first outputs a probability map which indicates how likely each voxel lies on a major sulcal curve. Next, it applies dynamic programming to extract the best curve based on the probability map and a shape prior. The algorithm has almost no parameters to tune for extracting different major sulci. It is very fast (it runs in under 1 min per sulcus including the time to compute the discriminative models) due to efficient implementation of the features (e.g., using the integral volume to rapidly compute the responses of 3-D Haar filters). Because the algorithm can be applied to MRI volumes directly, there is no need to perform preprocessing such as tissue segmentation or mapping to a canonical space. The learning aspect of our approach makes the system very flexible and general. For illustration, we use volumes of the right hemisphere with several major cortical sulci manually labeled. The algorithm is tested on two groups of data, including some brains from patients with Williams Syndrome, and the results are very encouraging. PMID- 17427742 TI - Classification based on cortical folding patterns. AB - We describe here a classification system based on automatically identified cortical sulci. Multivariate recognition methods are required for the detection of complex brain patterns with a spatial distribution. However, such methods may face the well-known issue of the curse of dimensionality-the risk of overfitting the training dataset in high-dimensional space. We overcame this problem, using a classifier pipeline with one- or two-stage of descriptor selection based on machine-learning methods, followed by a support vector machine classifier or linear discriminant analysis. We compared alternative designs of the pipeline on two different datasets built from the same database corresponding to 151 brains. The first dataset dealt with cortex asymmetry and the second dealt with the effect of the subject's sex. Our system successfully (98%) distinguished between the left and right hemispheres on the basis of sulcal shape (size, depth, etc.). The sex of the subject could be determined with a success rate of 85%. These results highlight the attractiveness of multivariate recognition models combined with appropriate descriptor selection. The sulci selected by the pipeline are consistent with previous whole-brain studies on sex effects and hemispheric asymmetries. PMID- 17427743 TI - Weighted fourier series representation and its application to quantifying the amount of gray matter. AB - We present a novel weighted Fourier series (WFS) representation for cortical surfaces. The WFS representation is a data smoothing technique that provides the explicit smooth functional estimation of unknown cortical boundary as a linear combination of basis functions. The basic properties of the representation are investigated in connection with a self-adjoint partial differential equation and the traditional spherical harmonic (SPHARM) representation. To reduce steep computational requirements, a new iterative residual fitting (IRF) algorithm is developed. Its computational and numerical implementation issues are discussed in detail. The computer codes are also available at http://www.stat.wisc.edu/ mchung/softwares/weighted.SPHARM/weighted-SPHARM.html. As an illustration, the WFS is applied i n quantifying the amount ofgray matter in a group of high functioning autistic subjects. Within the WFS framework, cortical thickness and gray matter density are computed and compared. PMID- 17427744 TI - Cortical surface shape analysis based on spherical wavelets. AB - In vivo quantification of neuroanatomical shape variations is possible due to recent advances in medical imaging and has proven useful in the study of neuropathology and neurodevelopment. In this paper, we apply a spherical wavelet transformation to extract shape features of cortical surfaces reconstructed from magnetic resonance images (MRIs) of a set of subjects. The spherical wavelet transformation can characterize the underlying functions in a local fashion in both space and frequency, in contrast to spherical harmonics that have a global basis set. We perform principal component analysis (PCA) on these wavelet shape features to study patterns of shape variation within normal population from coarse to fine resolution. In addition, we study the development of cortical folding in newborns using the Gompertz model in the wavelet domain, which allows us to characterize the order of development of large-scale and finer folding patterns independently. Given a limited amount of training data, we use a regularization framework to estimate the parameters of the Gompertz model to improve the prediction performance on new data. We develop an efficient method to estimate this regularized Gompertz model based on the Broyden-Fletcher-Goldfarb Shannon (BFGS) approximation. Promising results are presented using both PCA and the folding development model in the wavelet domain. The cortical folding development model provides quantitative anatomic information regarding macroscopic cortical folding development and may be of potential use as a biomarker for early diagnosis of neurologic deficits in newborns. PMID- 17427745 TI - Multiscale 3-D shape representation and segmentation using spherical wavelets. AB - This paper presents a novel multiscale shape representation and segmentation algorithm based on the spherical wavelet transform. This work is motivated by the need to compactly and accurately encode variations at multiple scales in the shape representation in order to drive the segmentation and shape analysis of deep brain structures, such as the caudate nucleus or the hippocampus. Our proposed shape representation can be optimized to compactly encode shape variations in a population at the needed scale and spatial locations, enabling the construction of more descriptive, nonglobal, nonuniform shape probability priors to be included in the segmentation and shape analysis framework. In particular, this representation addresses the shortcomings of techniques that learn a global shape prior at a single scale of analysis and cannot represent fine, local variations in a population of shapes in the presence of a limited dataset. Specifically, our technique defines a multiscale parametric model of surfaces belonging to the same population using a compact set of spherical wavelets targeted to that population. We further refine the shape representation by separating into groups wavelet coefficients that describe independent global and/or local biological variations in the population, using spectral graph partitioning. We then learn a prior probability distribution induced over each group to explicitly encode these variations at different scales and spatial locations. Based on this representation, we derive a parametric active surface evolution using the multiscale prior coefficients as parameters for our optimization procedure to naturally include the prior for segmentation. Additionally, the optimization method can be applied in a coarse-to-fine manner. We apply our algorithm to two different brain structures, the caudate nucleus and the hippocampus, of interest in the study of schizophrenia. We show: 1) a reconstruction task of a test set to validate the expressiveness of our multiscale prior and 2) a segmentation task. In the reconstruction task, our results show that for a given training set size, our algorithm significantly improves the approximation of shapes in a testing set over the Point Distribution Model, which tends to oversmooth data. In the segmentation task, our validation shows our algorithm is computationally efficient and outperforms the Active Shape Model algorithm, by capturing finer shape details. PMID- 17427746 TI - Anatomical equivalence class: a morphological analysis framework using a lossless shape descriptor. AB - Methods of computational anatomy are typically based on a spatial transformation that maps a template to an individual anatomy and vice versa. However, important morphological characteristics are frequently not captured by this transformation, thereby leading to lossy representations. We extend this formulation by incorporating residual anatomical information, i.e., information that is not captured by the shape transformation but is necessary in order to fully and exactly reconstruct the anatomy under measurement. We, therefore, arrive at a lossless morphological representation. By virtue of being lossless, this representation allows us to represent the same anatomy by an infinite number of pairs [transformation, residual], since different residuals correspond to different transformations. We treat these pairs as members of an anatomical equivalence class (AEC), which we approximate using principal component analysis. We show that projection onto the orthogonal to the AEC subspace produces measurements that allow us to better detect morphological abnormalities by eliminating variation in the data that is irrelevant and confounds underlying subtle morphological characteristics. Finally, we show that higher classification rates between a group of normal brains and a group of brains with localized atrophy are obtained if we use nonmetric distances between AECs instead of conventional Euclidean distances between individual morphological measurements. The results confirm that this representation can improve the results compared to conventional analysis, but also highlight limitations of the current approach and point to directions of further development of this general morphological analysis framework. PMID- 17427747 TI - [Effects of trace elements on the immune system]. AB - Trace elements as essential micronutrients play an important role in various physiological processes and are crucial for proper functioning of the immune system. Deficiency of trace elements and infectious diseases are often concomitantly observed and result in complex interactions. The major trace elements such as selenium, zinc, copper, manganese, etc. have immunomodulatory effects and thus influence susceptibility to and the course and outcome of a variety of viral infections. Some trace elements inhibit viral replication in the host cells and therefore have antiviral activity. Many trace elements act as antioxidants or are able not only to regulate the host immune response but also to alter the viral genome. This article gives a brief review of the major interactions between the trace elements and the immune system. PMID- 17427748 TI - [Tetanus immunity in the elderly in the Czech Republic]. AB - The study objective was to test immunity against tetanus in the elderly in the Czech Republic. Within serological surveys and in old people's homes, 776 persons aged 60 years or more were tested for antibodies against tetanus by an ELISA method. As many as 91 % of these persons had medium to high titers of antibodies against tetanus. Low but presumably still protective titers of antibodies against tetanus were detected in 9 % of the population tested. We documented good immunity against tetanus among the elderly in the Czech Republic as a result of a rich immunization history in this country as compared with some other countries in the world. The sporadic incidence of tetanus cases in the Czech Republic over the last 15 years indirectly indicates the efficacy of the immunization strategy used. PMID- 17427749 TI - [Microbial biofilms in the food industry]. AB - Biofilms are microbial communities whose architecture includes microorganisms, biotic substances produced by these microorganisms and attached organic and inorganic substances from the environment. They pose a serious problem in human medicine. Microbial biofilm communities are also cause for concern in the food industry since pathogenic microorganisms released from the biofilm may contaminate food and raw materials for food production. Not only the microbial community as a whole but also particular cells exhibit increased resistance to sanitation measures and disinfectants which makes it difficult to remove the biofilm or to inactivate particular built up microorganisms. Mainly model studies and molecular genetic and microscopy methods can contribute to better understanding of this issue, and thus to prevention and inactivation of microbial communities on food contact surfaces of equipment in the food production plants. Such studies would be of benefit to both health care and food processing and production. PMID- 17427750 TI - [What are the prospects for medical microbiology in the next 20 years?]. AB - In the future, microbiological laboratories are expected to maintain their proficiency in both molecular biological procedures and conventional methods. PMID- 17427751 TI - [Notified cases of imported transmisssible infections in the Czech Republic]. AB - OBJECTIVES: Based on analysis of epidemiological characteristics, to determine trends in the incidence of imported transmissible diseases in the Czech Republic and to draw attention to the epidemiological significance of international tourism and migration. MATERIAL AND METHODS: In cooperation with the National Reference Centre for Epidemiological Data Analysis, National Institute of Public Health Prague, the nationwide Epidat information system databases from 1993 to 2005 were analyzed using software Epi Info version 6.04d. RESULTS: Between 1993 and 2005, altogether 12,091 cases of transmissible diseases were imported into the Czech Republic from 168 countries in the world. The notified annual incidence of imported infections ranges from 206 to 1,714 cases. The highest percentage of cases was imported by Czech tourists (50.7 %) while 40 % of the notified cases were imported into the Czech Republic by foreigners. The period 1998-2004 for which the data on Czech tourism to other countries are available is characterized by increase in the absolute number of imported cases while the relative incidence of imported cases per 100,000 population traveling to other countries shows a downward trend. The highest numbers of cases were imported from Viet Nam (1,258 cases), Slovakia (1,155 cases) and India (786 cases). When considering Czech tourism to other countries for which the corresponding data are available, the highest rates of imported diseases in 1998-2004 were linked to the travels to Tunisia (1.18 imported cases per 1,000 tours), Bulgaria (0.69 per 1,000 tours) and Turkey (0.65 per 1,000 tours). The leading diagnosis of imported cases was salmonellosis (22%), followed by campylobacteriosis (10%) and trichuriasis (8%). CONCLUSION: Analysis of the Epidat information system databases for reporting transmissible diseases enables assessment of trends in imported infections in the Czech Republic as a point of departure for estimation of health risk from tourism and migration. PMID- 17427752 TI - [Failures to comply with the routine childhood immunization schedule due to contraindications and the use of alternative vaccines in children aged 0-4 years in the Czech Republic]. AB - The study objective was to determine the rate of failures to present for routine childhood immunization due to either parental vaccine refusal, failure to comply with the immunization schedule and real or false contraindications. The rate of use of alternative vaccines which can be provided within the routine immunization schedule either on parental request or when recommended by the physician was also determined. We analyzed records of 5,038 children born between January 1, 2000 and December 31, 2004 and registered with a sample of general practitioners. We identified 1,284 cases (25.5 %) of no, incomplete or alternative vaccination. Failure to present for immunization with at least one vaccine as scheduled due to contraindication was stated in 291 (5.8 %) children; a total of 436 contraindications were reported (more than 1 contraindication in some children). The most common contraindication was a CNS disorder (171 cases) such as encephalopathy, epilepsy or unspecified conditions. Sixty-nine children (1.4 %) remained nonvaccinated against at least one disease (due to parental vaccine refusal or failure to comply with the immunization schedule). Alternative vaccines were administered to 936 (18.5 %) children for the following reasons: contraindication in 275 (5.5 %) children and on parental request in 716 (14.2 %) children (cost incurred by parents), with 55 (1.1 %) children given alternative vaccines for both reasons. These results suggest that contraindications and the use of alternative vaccines are quite common and need to be monitored. PMID- 17427753 TI - [Pancreas cancer epidemiology]. AB - Pancreas cancer is characterized by a rapid and fatal outcome. It is a non communicable disease showing increasing incidence mainly in Europe, North America and Japan. In the Czech Republic, the incidence of pancreas cancer has more than tripled over the last 50 years. The pancreas cancer incidence and mortality rates are practically identical in this country. The standardized incidence rates (world standard) are 10.4 in men and 7.1 in women, with the Czech Republic ranking the sixth and fourth in the world, respectively. The standardized mortality rates are 10.4 and 7.2, respectively, with the Czech Republic ranking the fourth and sixth to seventh in the world, respectively. Very rapid fatality is also indicative of the seriousness of the disease whose median survival ranges from three to six months. PMID- 17427754 TI - The role of FB21 as a marker of glomerular endothelial cell injury. AB - FB21 was reactive with the glomerular endothelial cells and distal tubules of the human kidney and was bound to a sialic-acid-dependent cell surface antigen. We evaluated the FB21 staining in fetal kidneys, and the kidneys of children and adults with normal kidneys and glomerulonephritis and investigated whether FB21 can be used as a marker for endothelial cell injury. FB21 was reactive with the endothelial cells of normal kidneys and was detected on the surface of endothelial cells by immunoelectron microscopy. FB21 was reactive with endothelial cells in the kidneys of over 32-week fetuses. The endothelial cell FB21 staining scores in the first renal biopsy specimens of patients with hemolytic uremic syndrome (HUS) were lower than in the kidneys of children with normal kidneys and was negatively correlated with their serum E-selectin concentrations. The FB21 staining of glomerular endothelial cells was similar to the staining for the other endothelial markers, CD34 and von Willebrand factor (vWF). However, FB21 staining of interstitial blood vessels was very weak and was distinct from that of other endothelial markers. These results suggest that FB21 can be used as a specific marker for glomerular endothelial cell injury in various types of glomerulonephritis. PMID- 17427755 TI - Nitric oxide modulates biphasic contractile response of guinea pig vas deferens to electrical field stimulation. AB - Electrical field stimulation (EFS) produced a biphasic contractile response; viz. initial rapid phasic contraction and second slow tonic contraction, in isolated guinea pig vas deferens. Pretreatment with the substrate of nitric oxide (NO) synthase (NOS), 1 mM L-arginine (L-ARG), augmented both the initial rapid and the second slow contractile responses to EFS (5 Hz, 0.5 msec, 30 V, for 30 sec). The increase of stimulation frequency from 5 Hz to 10 Hz or 20 Hz tended to attenuate the augmented responses. On the contrary, pretreatment with an inhibitor of NOS, 0.1 mM NG-nitro-L-arginine (L-NNA) suppressed both the initial rapid and the second slow contractile responses to EFS. The suppressive effect on the initial rapid contraction was also attenuated by the increase of stimulation frequency from 5 Hz or 10 Hz to 20 Hz. Contractile response to exogenously administered 1 mM adenosine triphosphate (ATP) tended to be slightly increased and decreased by the treatment with 1 mM L-ARG and 0.1 mM L-NNA, respectively. Contractile response to exogenously administered 10 microM noradrenaline (NA) was almost unaffected by the treatment with 1 mM L-ARG, while the treatment with 0.1 mM L NNA slightly depressed the response. Potentiated contractile response to 1 mM ATP in the presence of 10 microM NA was further potentiated by the treatment with 1 mM L-ARG, while the response was almost unaffected by the treatment with 0.1 mM L NNA. These findings may indicate that NO acts mainly on presynaptic site and increases the release of chemical transmitter, ATP or prevents the inactivation of ATP. Also, NO may act, at least in part, on postsynaptic site and potentiates the contractile response to ATP in the presence of NA. PMID- 17427756 TI - Indoor-atmospheric radon-related radioactivity affected by a change of ventilation strategy. AB - The present author has kept observation for concentrations of atmospheric radon, radon progeny and thoron progeny for several years at the campus of Fukushima Medical University. Accidentally, in the midst of an observation term, i.e., February 2005, the facility management group of the university changed a strategy for the manner of ventilation, probably because of a recession: (I) tidy everyday ventilation of 7:30-24:00 into (II) shortened weekday ventilation of 8: 00-21 : 00 with weekend halts. This change of ventilation manner brought a clear alteration for the concentrations of radon-related natural radioactivity in indoor air. The present paper concerns an investigation of the effect of the ventilation strategy on the indoor-atmospheric radon-related radioactivity. PMID- 17427757 TI - Expression of human glucocorticoid receptor beta of peripheral blood mononuclear cells in patients with severe autoimmune hepatitis. AB - We evaluated the expression of human glucocorticoid receptor beta (hGRbeta) in patients with severe autoimmune hepatitis (AIH). The subjects were 27 patients with AIH, including 6 with severe type (prothrombin time [PT] < 40%) and 21 with non-severe type (PT>40%). Total RNA extracted from peripheral blood mononuclear cells (PBMCs) was reversed using reverse transcriptase. The resultant complementary DNA was amplified by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for hGR alpha and beta. The six patients with severe AIH were female; three presented fulminant hepatic failure with hepatic encephalopathy. In all patients with AIH, hGR a was detected. The incidence of hGR beta expression in patients with non-severe type was 42.9% (9/21) ; it was 100% (6/6) in those with severe type. The positive ratio was significantly higher in severe-type patients. These results suggest that hGR beta expression in PBMCs is a novel predictor of AIH severity. PMID- 17427758 TI - Clinicolaboratory characteristics of Japanese patients with primary biliary cirrhosis-autoimmune hepatitis overlap. AB - To clarify the clinicolaboratory characteristics of patients with primary biliary cirrhosis (PBC)-autoimmune hepatitis (AIH) overlap, we analyzed their clinicolaboratory findings and compared them with those of patients with AIH or PBC retrospectively. We analyzed the laboratory findings of 177 patients that diagnosed 103 PBC and 74 AIH patients at our department during the period from January 1990 to April 2005. Of 103 PBC patients with a diagnosis of PBC, we identified 10 cases (9.7%) of PBC-AIH overlap (2 male, 8 female; mean age 56.5 years). PBC preceded AIH in 2 patients, and both diseases occurred simultaneously in the other 8 patients. There is no patients AIH preceded PBC. Positive frequency of anti-smooth muscle antibody (ASMA), IgG and IgM levels were significantly higher in patients with overlap than in those with AIH or PBC. Ursodeoxychoric acid (UDCA) was administered to all 10 patients initially, and later an immunosuppressant, prednisolone or azathioprine, was added in 6 patients. Two of the 10 patients died of liver failure 5 and 12 years after diagnosis, respectively. Both patients had been treated by either prednisolone or UDCA alone. We conclude that in patients with PBC-AIH overlap, the clinical characteristics of both PBC and AIH exist in an enhanced manner. PMID- 17427759 TI - Possible association of cytotoxic T lymphocyte antigen-4 genetic polymorphism with liver damage of primary biliary cirrhosis in Japan. AB - Cytotoxic T lymphocyte antigen-4 (CTLA-4) is an important inhibitor of T lymphocyte response. Polymorphisms in the CTLA-4 gene have been reported to be associated with numerous autoimmune diseases. The aim of this study was to determine whether polymorphisms of CTLA-4 exon 1 (+49) genes are associated with susceptibility and clinicolaboratory findings of primary biliary cirrhosis (PBC) in the Japanease population. Blood samples were obtained from 45 patients (6 men and 39 women, aged 23-56 years) with PBC and 73 healthy controls (48 men and 25 women, aged 22-72 years). CTLA-4 exon 1 (+49) polymorphism was defined using a polymerase chain reaction-restriction fragment length polymorphism with Bst71I restriction enzyme. The genotype frequencies of A/A, A/G, and G/G in 45 patients with PBC were 11% (5 patients), 44% (20 patients), and 44% (20 patients), respectively. There was no significant difference between frequencies in PBC patients and healthy controls. PBC patients with G/G genotype had significantly higher serum levels of ALT, GGT, and IgM than those in patients with A/A or A/G genotype. In conclusion, CTLA-4 gene polymorphisms are not associated with susceptibility of PBC in Japan; however, G/G genotype may be associated with liver damage. PMID- 17427760 TI - Estimation of microinhomogeneity of conduction impairment by wavelet analysis during early phase of myocardial ischemia in pigs. AB - Ventricular fibrillation (VF) is most frequent in the very early phase in acute coronary occlusion, and is triggered by the re-entrant mechanism in this phase. An inhomogeneous conduction in the ischemic myocardium would be substrates for re entry. The aim of this study was to examine the relationship between the severity of irregularities of the QRS complex and VF. Eleven pigs were analyzed, and the heart was fixed in the pericardial cradle. Ag-AgCl bipolar electrodes were fixed on the epicardium in ischemic and non-ischemic regions. The proximal portion of the left anterior descending coronary artery was occluded for one hour. Electrocardiograms (ECGs) were continuously recorded on a magnetic tape, and wavelet analysis was performed on signal-averaged ECG (25 beats) every 60 sec after the experiment. The number of local maxima (N) and the duration between the first and the last local maximum (D) were automatically measured. N and D significantly increased in the ischemic area, but not in the non-ischemic area. N and D increased approximately twofold just before the occurrence of VF in 8 fibrillated pigs (p<0.01, each). There were significant positive linear relationships between the rate of increase in N and D to VF and basal heart rate before coronary occlusion (r=0.90, p <0.01 in N, r=0.84, p <0.01 in D at 160 Hz). These results suggest that there would be a threshold inhomogeneous conduction for the occurrence of VF and an increase in heart rate would accelerate the inhomogeneous conduction in acute myocardial ischemia. PMID- 17427761 TI - Usefulness of complement split product, Bb, as a clinical marker for disease activity of lupus nephritis. AB - To evaluate the usefulness of Bb, a split product of complement factor B, as a clinical marker for disease activity of lupus nephritis, we measured the Bb concentration of sera from 42 patients with lupus nephritis. Serum Bb levels were significantly higher in patients with active nephritis (active nephritis group, n= 30) than in patients with nephritis in remission (remission group, n=12) (14.3+/-8.3 versus 7.4+/-5.9 microg/ml; p = 0.012). In contrast, there was no significant difference in serum C3 levels between active nephritis group and remission group (42.5+/-20.9 versus 44.7+/-15.9 mg/dl ; p = 0.77). In the comparison of Bb levels between active nephritis group and remission group, the sensitivity was 66.6%, specificity was 83.3%, and the positive and negative likelihood ratios were 3.95% and 0.41%, respectively. The present results suggest that serum Bb level is a useful clinical marker for disease activity in lupus nephritis. PMID- 17427762 TI - Effect of isosorbide dinitrate on gastric blood flow in rats with liver cirrhosis determined by analyzing gastric blood flow, portal vein pressure and blood gas. AB - We investigated the effects of isosorbide dinitrate (IDN) on gastric blood flow (GBF), portal venous pressure (PVP) and blood gas of rats with liver cirrhosis (LC) accompanied by portal hypertension. Thirty male Wistar rats (LC in 17 and normal in 13) were used. Before and after IDN administration, GBF, PVP and blood gas in the femoral artery and portal vein were measured. Portal blood oxygen concentration was estimated by calculating the ratio of PO2 in portal blood and that in arterial blood (PpvO2/PaO2) of each rat. The GBF in the LC rats was significantly lower than that in the normal rats. In the LC group, IDN administration significantly increased the GBF. The PpvO2/PaO2 value in the group with LC was significantly lower after IDN administration than that before IDN administration. In the investigation whether changes in PVP or Ppv/PaO2 contributed more to the change in GBF after IDN administration, a significant correlation was found between rates of change in GBF and PpvO2/PaO2 were significantly correlated (r= -0.733, p <0.05). The effect of IDN on changes in the stomach accompanying portal hypertension is mainly attributable to a decrease in preload, which suppresses inflow to the stomach, as reflected by a decrease in PpvO2/ PaO2, rather than to a decrease in afterload on GBF, as reflected by a decrease in PVP. PMID- 17427763 TI - Suppressive effect of acupuncture stimulation to the sacral segment on the state of vigilance and the brainstem cholinergic neurons. AB - The effects of acupuncture stimulation to the sacral segment on the electroencephalogram (EEG) and activity of the cholinergic neurons in the laterodorsal tegmental nucleus (LDT) were examined in urethane-anesthetized rats. When EEG was small amplitude and higher frequency, the stimulation to the sacral segment induced large amplitude and slow EEG with latencies ranged from 45 sec to 12 min, and durations from 48 sec to 56 min. The stimulus induced EEG is composed of significant increase in delta power and significant decrease in theta and beta powers. Firing rate of the cholinergic LDT neurons significantly decreased from 2.9+/-1.5 Hz to 1.1+/-0.8 Hz after the stimulus (n=12, p<0.05). The decrease of neuronal activity always preceded to the start of large and slow EEG, while the increase of the activity always preceded to the change of EEG from large slow wave to small faster wave. These results suggest that the acupuncture stimulation to the sacral segment changes the state of the animals from light anesthesia to deep anesthesia, and that the change is mediated by the suppression of the cholinergic neurons in the LDT. PMID- 17427764 TI - Case of an elderly man with associated Henoch-Schonlein purpura during treatment of acute pancreatitis. AB - A 70-year-old man with acute pancreatitis (acute exacerbation of chronic pancreatitis) was admitted to our department. Despite temporary improvement, the pancreatitis worsened on the 21st hospital day, forming a pancreatic pseudocyst, with infection in the cyst. After treatment with various antibiotics, a blood test on the 71st hospital day indicated improved inflammatory response despite continuing abdominal cramps. From the 75th hospital day, the patient developed purpura and arthralgia of the lower limbs, with melena and hematuria. Henoch Schonlein purpura was diagnosed definitively by skin biopsy. Such a complication of acute pancreatitis with Henoch-Schonlein purpura is rare. This case also suggests that microvasculitis around the pancreas resulting from Henoch-Schonlein purpura might have prolonged the pancreatitis. PMID- 17427765 TI - Effects of fluvoxamine on behavioral and psychological symptoms of dementia in Alzheimer's disease: a report of three cases. AB - OBJECTIVE: To report 3 cases of severe behavioral and psychological symptoms of dementia (BPSD) in Alzheimer's disease (AD) with fluvoxamine treatment and to discuss the treatment implications for use of the drug. CASE SUMMARY: An 83-year old woman was diagnosed with AD. Before treatment, she showed sudden irritation and excitement. Her BEHAVE-AD score was 40. She was started on fluvoxamine and quetiapine. Eight weeks later, she was friendly and thankful towards the staff. Her BEHAVE-AD score was 10. The second case was a 79-year-old woman diagnosed with AD. Before treatment, she attempted to leave our hospital and wandered and shouted throughout the day. Her BEHAVE-AD score was 42. She was started on fluvoxamine, and the dosage was gradually increased. Eight weeks later, the shouting and excitement disappeared almost completely. Her BEHAVE-AD score was 13. The third case was a 79-year-old man diagnosed with AD. Before treatment, we put him in a private, locked room because he was extremely agitated and violent because of delusions. His BEHAVE-AD score was 42. He was started on fluvoxamine and sodium valproate. Eight weeks later, the delusion became mild and did not affect his mood or behavior. His BEHAVE-AD score at this point was 4. DISCUSSION: Fluvoxamine was effective in controlling BPSD with AD. This finding shows that the pathophysiology of BPSD due to AD may occur because of a hyposerotonergic state in the brain. CONCLUSION: These cases show that fluvoxamine appears to be effective in the control of BPSD with AD. PMID- 17427766 TI - A case of rapidly expanding and increasing focal nodular hyperplasia. AB - We report a case of 14-year-old male patient who underwent bile-duct-to-jejunum anastomosis because of congenital biliary atresia at the age of 2 months. A 15 mm hypervascular nodule was detected for the first time in the S1 region of the liver at the age of 9 years. Two years later, 6 hypervascular nodules were found in the liver. A tumor biopsy was performed. It was diagnosed as a focal nodular hyperplasia (FNH). However, the number of nodules increased from 6 to 12 and those diameters were enlarged two to seven times one year later; the tumor biopsy was performed again. Histologically, the findings were consistent with those obtained previously, which indicated FNH. We consider that this is a very rare case of FNH in which both the number of nodules and the size were increased in a short period of time. We present it here as a valuable case report. PMID- 17427767 TI - Stapes mobilization revisited: primum non nocere. PMID- 17427768 TI - Jugular bulb diverticulum. PMID- 17427769 TI - Endoscopic view of a powered partial resection of a concha bullosa. PMID- 17427770 TI - Teflon granuloma. PMID- 17427771 TI - Skull base thrombotic mycosis. PMID- 17427772 TI - Ramsay Hunt syndrome, type I. PMID- 17427773 TI - Management of paratracheal adenoid cystic carcinoma. PMID- 17427774 TI - Correction of caudal septal deviation: use of a caudal septal extension graft. PMID- 17427775 TI - Pay-for-call reimbursement: what's your experience? PMID- 17427776 TI - Psychological characteristics of patients with Meniere's disease compared with patients with vertigo, tinnitus, or hearing loss. AB - An association between Meniere 's disease and psychological distress is frequently reported. Patients who do not have Meniere's disease but who have similar symptoms also experience various kinds of psychological disturbances. We conducted a study to investigate the relationship between Meniere's disease and personality traits, illness behavior, depression, and anxiety. We compared these factors in 77 patients who had Meniere's disease and 133 controls who did not have the disease but had one of its symptoms--either vertigo, tinnitus, or hearing loss. The mental status of study participants was assessed with standard tests. Patients in both groups had higher than normal levels of anxiety and neuroticism. The only significant difference between the two groups was a higher rate of extroversion in the Meniere's disease group. Minor differences emerged when Meniere's patients with tinnitus or vertigo were compared with similar controls. Relationships between psychological observations and otologic symptomatology or an otologic diagnosis were not specific, which illustrates the need to consider the role of illness behavior and personality as targets for psychological support or therapy associated with ENT treatment. PMID- 17427777 TI - The role of endoscopic sinus surgery in the diagnosis and treatment of metastatic orbital carcinoid tumors. AB - Carcinoid tumor metastatic to the orbit is a rare occurrence. We report such a case in a patient with a carcinoid metastasis that was found in the medial rectus muscle. The diagnosis was established via a transnasal endoscopic approach. We also discuss the treatment of metastatic orbital carcinoid in the hope that we will increase awareness of the utility of transnasal endoscopy in facilitating both the diagnosis and treatment of posteromedial orbital pathology. PMID- 17427778 TI - Bilateral peritonsillar abscesses: a challenging diagnosis. AB - Peritonsillar abscess is the most common complication of acute tonsillitis. Bilateral peritonsillar abscesses are much less common, and they may be more difficult to detect on physical examination because the oropharynx often appears to be symmetrical rather than asymmetrical, as is the case in unilateral abscess. Previous steroid treatment may also complicate the diagnosis by masking the signs and symptoms of abscess. We describe the case of a young woman who presented to the emergency department with relatively mild symptoms despite having large bilateral peritonsillar abscesses. We believe that her symptoms had been masked by previous steroid therapy. We also review the treatment and microbiology of peritonsillar abscess. PMID- 17427779 TI - Acute prevertebral calcific tendinitis: a nonsurgical cause of prevertebral fluid collection. AB - Calcific tendinitis of the prevertebral muscles is a rare clinical entity. Its nonspecific presenting symptoms (e.g., retropharyngeal space infection) may mimic an infectious etiology. Treatment is based on the administration of a nonsteroidal anti-inflammatory drug (NSAID) for 2 to 3 weeks and cessation of symptom-provoking activity. Most patients will improve greatly within the first 48 to 72 hours after the initiation of an NSAID, and complete resolution generally occurs within 3 weeks. We describe a new case of prevertebral calcific tendinitis, and we review the literature on this condition. PMID- 17427780 TI - Irreversible hydrocolloid: an unusual presentation of esophageal obstruction. AB - Upper digestive tract obstruction can occur after ingestion of various types of foreign bodies. However to the best of our knowledge, no case of a near-total obstruction caused by an irreversible hydrocolloid has heretofore been reported in the literature. We present just such a case, and we discuss our preferred method of removing foreign bodies from the cervical esophagus. PMID- 17427781 TI - Extent of training crisis revealed. PMID- 17427782 TI - 'The impact of blood cancers has been systematically underestimated'. PMID- 17427783 TI - To strike or not to strike. PMID- 17427784 TI - The nurse who drove down obesity. PMID- 17427785 TI - NICE guidance on antenatal and postnatal mental health. PMID- 17427786 TI - Reproductive system: part one--male anatomy. AB - This article, the first in a four-part series, explores the core anatomy of the male human reproductive system. Part two will focus on its physiology and function. Part three and four will explore the female human reproductive anatomy and its reproductive physiology. The series should enhance the reader's theory and practice when caring for patients with reproductive or sexual health needs. PMID- 17427787 TI - Protecting children from all forms of abuse and neglect. AB - Nurses have a key role in safeguarding children from abuse and neglect. This article outlines nurses' responsibilities in child protection and general protocols for talking to children and reporting concerns about their welfare. PMID- 17427788 TI - Infection protection. State health departments look to the national healthcare safety network to help with infection reporting mandates. PMID- 17427790 TI - ELINCS gets adopted. HL 7 is taking over ELINCS to ensure lab data and EHRs continue towards interoperability. PMID- 17427789 TI - Certification commission charts future. CCHIT's Mark Leavitt lays out his organization's plans to keep certification efforts moving. PMID- 17427791 TI - Receivables financing: 101. When everything else has been mortgaged to the hilt, there's one more place to go for solvency. PMID- 17427792 TI - E-prescribing for all. A new industry group is looking to remove the technological and financial barriers that keep physicians from e-prescribing. PMID- 17427793 TI - Now it's personal. Personal health records may be next on deck for the certification process. PMID- 17427794 TI - Giants join forces. Bringing together the VA's inpatient system and DoD's outpatient EMR may serve as a prime example of cooperation. PMID- 17427795 TI - Image is everything. No longer solely the domain of radiologists, imaging is taking hold throughout the hospital. PMID- 17427796 TI - Guarding the perimeter. New security risks and HIPAA compliance continue to challenge IT professionals. PMID- 17427797 TI - Crosswalking the NPI chasm, part II. Any NPI strategy must clearly spell out both solutions and results. PMID- 17427798 TI - What's in a name? Clarity and common sense can go a long way when setting up master data files. PMID- 17427799 TI - Practice moves full speed ahead. With a new practice management system, Farragut Family Practice masters the management of business operations. PMID- 17427800 TI - Payer buys IT to improve. Dean Health Plan enhances medical management capabilities for improved satisfaction and decreased costs. PMID- 17427801 TI - Securing the network. Network access control solutions can be an important tool in the fight against intrusion. PMID- 17427802 TI - Contract for success. San Jose Medical Group has inked a deal with MPV Inc. that will help ensure its payers live up to their commitments. PMID- 17427803 TI - Getting up and running. Englewood Hospital and Medical Center found a low-cost approach to bringing new employees on board. PMID- 17427804 TI - ABA is required for Leptosphaeria maculans resistance via ABI1- and ABI4 dependent signaling. AB - Abscisic acid (ABA) is a defense hormone with influence on callose-dependent and independent resistance against Leptosphaeria maculans acting in the RLMcol pathway. ABA-deficient and -insensitive mutants in Ler-0 background (abal-3 and abil-1) displayed susceptibility to L. maculans, along with a significantly decreased level of callose depositions, whereas abi2-1 and abi3-1 remained resistant, together with the abi5-1 mutant of Ws-0 background. Suppressor mutants of abil-1 confirmed that the L. maculans-susceptible response was due to the dominant negative nature of the abil-1 mutant. Highly induced camalexin levels made ABA mutants in Col-0 background (aba2-1, aba3-1, and abi4-1) appear resistant, but displayed enhanced susceptibility as double mutants with pad3-1, impaired in camalexin biosynthesis. beta-Aminobutyric acid (BABA) pretreatment of Ler-0 contributed to an elevated level of endogenous ABA after L. maculans inoculation. Comparisons between (RLM1co1)pad3 and rlmlLerpad3 showed that ABA and BABA enhancement of callose deposition requires induction from RLM1col. ABII, but not ABI2, was found to be involved in a feedback mechanism that modulates RLM1co, expression. Genetic analysis showed further that this feedback occurs upstream of ABI4 and that components downstream of ABI4 modulate ABIJ activity. ABA and BABA treatments of the L. maculans-susceptible callose synthase mutant pmr4 showed that ABA also induces a callose-independent resistance. Similar treatments enhanced callose depositions and induced resistance to L. maculans in oilseed rape, and BABA-induced resistance was found to be independent of salicylic acid. PMID- 17427805 TI - The HopX (AvrPphE) family of Pseudomonas syringae type III effectors require a catalytic triad and a novel N-terminal domain for function. AB - Many gram-negative plant pathogenic bacteria employ type III secretion systems to deliver effector proteins directly into the host cell during infection. On susceptible hosts, type III effectors aid pathogen growth by manipulating host defense pathways. On resistant hosts, some effectors can activate specific host disease resistance (R) genes, leading to generation of rapid and effective immune responses. The biochemical basis of these processes is poorly understood. The HopX (AvrPphE) family is a widespread type III effector among phytopathogenic bacteria. We determined that HopX family members are modular proteins composed of a conserved putative cysteine-based catalytic triad and a conserved potential target/cofactor interaction domain. HopX is soluble in host cells. Putative catalytic triad residues are required for avirulence activity on resistant bean hosts and for the generation of a cell-death response in specific Arabidopsis genotypes. The putative target/cofactor interaction domain is also required for these activities. Our data suggest that specific interaction with and modification of a cytosolic host target drives HopX recognition in resistant hosts and may contribute to virulence in susceptible hosts. Surprisingly, the Legionella pneumophila genome was found to contain a protein with similarity to HopX in sequence and domain arrangement, suggesting that these proteins might also contribute to animal pathogenesis and could be delivered to plant and animal hosts by diverse secretion systems. PMID- 17427806 TI - Spatial analysis of arabidopsis thaliana gene expression in response to Turnip mosaic virus infection. AB - Virus-infected leaf tissues comprise a heterogeneous mixture of cells at different stages of infection. The spatial and temporal relationships between sites of virus accumulation and the accompanying host responses, such as altered host gene expression, are not well defined. To address this issue, we utilized Turnip mosaic virus (TuMV) tagged with the green fluorescent protein to guide the dissection of infection foci into four distinct zones. The abundance of Arabidopsis thaliana mRNA transcripts in each of the four zones then was assayed using the Arabidopsis ATH1 GeneChip oligonucleotide microarray (Affymetrix). mRNA transcripts with significantly altered expression profiles were determined across gradients of virus accumulation spanning groups of cells in and around foci at different stages of infection. The extent to which TuMV-responsive genes were up- or downregulated primarily correlated with the amount of virus accumulation regardless of gene function. The spatial analysis also allowed new suites of coordinately regulated genes to be identified that are associated with chloroplast functions (decreased), sulfate assimilation (decreased), cell wall extensibility (decreased), and protein synthesis and turnover (induced). The functions of these downregulated genes are consistent with viral symptoms, such as chlorosis and stunted growth, providing new insight into mechanisms of pathogenesis. PMID- 17427807 TI - Separate and combined disruptions of two exo-beta-1,3-glucanase genes decrease the efficiency of Pichia anomala (strain K) biocontrol against Botrytis cinerea on apple. AB - The modes of action of the antagonistic yeast Pichia anomala (strain K) have been studied; however, thus far, there has been no clear demonstration of the involvement of exo-beta-1,3-glucanase in determining the level of protection against Botrytis cinerea afforded by this biocontrol agent on apple. In the present study, the exo-beta-1,3-glucanase-encoding genes PAEXG1 and PAEXG2, previously sequenced from the strain K genome, were separately and sequentially disrupted. Transfer of the URA3-Blaster technique to strain K, allowing multiple use of URA3 marker gene, first was validated by efficient inactivation of the PaTRP1 gene and recovery of a double auxotrophic strain (uracil and tryptophan). The PAEXG1 and PAEXG2 genes then were inactivated separately and sequentially with the unique URA3 marker gene. The resulting mutant strains showed a significantly reduced efficiency of biocontrol of B. cinerea when applied to wounded apple fruit, the calculated protection level dropping from 71% (parental strain) to 8% (mutated strain) under some experimental conditions. This suggests that exo-beta-1,3-glucanases play a role in the biological control of B. cinerea on apple. Furthermore, biological control experiments carried out in this study underline the complexity of the host-antagonist-pathogen interaction. Two experimental parameters (yeast inoculum concentration and physiological stage of the fruit) were found to influence dramatically the protection level. Results also suggest that, under some conditions, the contribution of exo-beta-1,3 glucanase to biological control may be masked by other modes of action, such as competition. PMID- 17427808 TI - The consensus N-myristoylation motif of a geminivirus AC4 protein is required for membrane binding and pathogenicity. AB - Some geminiviruses encode a small protein, AC4, whose role in pathogenesis has only recently attracted attention. A few studies have shown that this protein is involved in pathogenesis and suppresses RNA silencing. Here, using Nicotiana benthamiana, we show that East African cassava mosaic Cameroon virus (EACMCV) AC4 is a pathogenicity determinant and that it suppresses the systemic phase of RNA silencing. Furthermore, confocal imaging analyses show that it binds preferentially to the plasma membrane as well as to cytosolic membranes including the perinucleus but is excluded from the nucleus. A computational examination of the AC4 protein encoded by the EACMCV, a bipartite geminivirus, shows that it encodes a consensus N-myristoylation motif and is likely posttranslationally myristoylated and palmitoylated. Replacement of Gly-2 and Cys-3 (sites of posttranslational attachment of myristic and palmatic acids, respectively) with alanine affected AC4 membrane binding and pathogenesis. Furthermore, replacement of Ile-5, a nonessential myristoylation residue, with alanine did not affect AC4 function. Together, these data indicate that EACMCV AC4 is likely dually acylated at Gly-2 and Cys-3 and that these modifications are intrinsic signals for membrane targeting and pathogenesis. This is the first report of a membrane protein to be involved in pathogenesis and RNA silencing suppression. PMID- 17427809 TI - A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction. AB - Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6, and mutant BcPG2-D192A caused no symptoms. Expression of BcPG2 caused the most severe symptoms, including wilting and necrosis. BcPG2 previously has been shown to be essential for B. cinerea virulence. The in vivo effect of this enzyme and the inhibition by a polygalacturonase-inhibiting protein (PGIP) was examined by coexpressing Bcpg2 and the Vvpgipl gene from Vitis vinifera in N. benthamiana. Coinfiltration resulted in a substantial reduction of the symptoms inflicted by the activity of BcPG2 in planta, as evidenced by quantifying the variable chlorophyll fluorescence yield. In vitro, however, no interaction between pure VvPGIP1 and pure BcPG2 was detected. Specifically, VvPGIP1 neither inhibited BcPG2 activity nor altered the degradation profile of polygalacturonic acid by BcPG2. Furthermore, using surface plasmon resonance technology, no physical interaction between VvPGIP1 and BcPG2 was detected in vitro. The data suggest that the in planta environment provided a context to support the interaction between BcPG2 and VvPGIP1, leading to a reduction in symptom development, whereas neither of the in vitro assays detected any interaction between these proteins. PMID- 17427810 TI - TolC is required for pathogenicity of Xylella fastidiosa in Vitis vinifera grapevines. AB - Xylella fastidiosa infects a wide range of hosts and causes serious diseases on some of them. The complete genomic sequences of both a citrus variegated chlorosis (CVC) and a Pierce's disease (PD) strain revealed two type I protein secretion plus two multidrug resistance efflux systems, and all evidently were dependent on a single tolC homolog. Marker exchange mutagenesis of the single tolC gene in PD strain Temecula resulted in a total loss of pathogenicity on grape. Importantly, the tolC- mutant strains were not recovered after inoculation into grape xylem, strongly indicating that multidrug efflux is critical to survival of this fastidious pathogen. Both survival and pathogenicity were restored by complementation using tolC cloned in shuttle vector pBBR1MCS-5, which was shown to replicate autonomously, without selection, for 60 days in Temecula growing in planta. These results also demonstrate the ability to complement mutations in X. fastidiosa. PMID- 17427811 TI - Xylella fastidiosa requires polygalacturonase for colonization and pathogenicity in Vitis vinifera grapevines. AB - Xylella fastidiosa is the causal agent of Pierce's disease of grape, an economically significant disease for the grape industry. X. fastidiosa systemically colonizes the xylem elements of grapevines and is able to breach the pit pore membranes separating xylem vessels by unknown mechanisms. We hypothesized that X. fastidiosa utilizes cell wall degrading enzymes to break down pit membranes, based on the presence of genes involved in plant cell wall degradation in the X. fastidiosa genome. These genes include several beta-1,4 endoglucanases, several xylanases, several xylosidases, and one polygalacturonase (PG). In this study, we demonstrated that the pglA gene encodes a functional PG. A mutant in pglA lost pathogenicity and was compromised in its ability to systemically colonize Vitis vinifera grapevines. The results indicate that PG is required for X. fastidiosa to successfully infect grapevines and is a critical virulence factor for X. fastidiosa pathogenesis in grapevine. PMID- 17427812 TI - Expression of AtWRKY33 encoding a pathogen- or PAMP-responsive WRKY transcription factor is regulated by a composite DNA motif containing W box elements. AB - WRKY transcription factors regulate distinct parts of the plant defense transcriptome. Expression of many WRKY genes themselves is induced by pathogens or pathogen-mimicking molecules. Here, we demonstrate that Arabidopsis WRKY33 responds to various stimuli associated with plant defense as well as to different kinds of phytopathogens. Although rapid pathogen-induced AtWRKY33 expression does not require salicylic acid (SA) signaling, it is dependent on PAD4, a key regulator upstream of SA. Activation of AtWRKY33 is independent of de novo protein synthesis, suggesting that it is at least partly under negative regulatory control. We show that a set of three WRKY-specific cis-acting DNA elements (W boxes) within the AtWRKY33 promoter is required for efficient pathogen- or PAMP-triggered gene activation. This strongly indicates that WRKY transcription factors are major components of the regulatory machinery modulating immediate to early expression of this gene in response to pathogen attack. PMID- 17427813 TI - The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca. AB - Podosphaera fusca is the main causal agent of cucurbit powdery mildew in Spain. Four Bacillus subtilis strains, UMAF6614, UMAF6619, UMAF6639, and UMAF8561, with proven ability to suppress the disease on melon in detached leaf and seedling assays, were subjected to further analyses to elucidate the mode of action involved in their biocontrol performance. Cell-free supernatants showed antifungal activities very close to those previously reported for vegetative cells. Identification of three lipopeptide antibiotics, surfactin, fengycin, and iturin A or bacillomycin, in butanolic extracts from cell-free culture filtrates of these B. subtilis strains pointed out that antibiosis could be a major factor involved in their biocontrol ability. The strong inhibitory effect of purified lipopeptide fractions corresponding to bacillomycin, fengycin, and iturin A on P. fusca conidia germination, as well as the in situ detection of these lipopeptides in bacterial-treated melon leaves, provided interesting evidence of their putative involvement in the antagonistic activity. Those results were definitively supported by site-directed mutagenesis analysis, targeted to suppress the biosynthesis of the different lipopeptides. Taken together, our data have allowed us to conclude that the iturin and fengycin families of lipopeptides have a major role in the antagonism of B. subtilis toward P. fusca. PMID- 17427814 TI - Iron acquisition from Fe-pyoverdine by Arabidopsis thaliana. AB - Taking into account the strong iron competition in the rhizosphere and the high affinity of pyoverdines for Fe(III), these molecules are expected to interfere with the iron nutrition of plants, as they do with rhizospheric microbes. The impact of Fe-pyoverdine on iron content of Arabidopsis thaliana was compared with that of Fe-EDTA. Iron chelated to pyoverdine was incorporated in a more efficient way than when chelated to EDTA, leading to increased plant growth of the wild type. A transgenic line of A. thaliana overexpressing ferritin showed a higher iron content than the wild type when supplemented with Fe-EDTA but a lower iron content when supplemented with Fe-pyoverdine despite its increased reductase activity, suggesting that this activity was not involved in the iron uptake from pyoverdine. A mutant knock-out iron transporter IRT1 showed lower iron and chlorophyll contents when supplemented with Fe-EDTA than the wild type but not when supplemented with Fe-pyoverdine, indicating that, in contrast to iron from EDTA, iron from pyoverdine was not incorporated through the IRT1 transporter. Altogether these data suggest that iron from Fe-pyoverdine was not incorporated in planta through the strategy I, which is based on reductase activity and IRT1 transporter. This is supported by the presence of pyoverdine in planta as shown by enzyme-linked immunosorbent assay and by tracing 15N of 15N-pyoverdine. PMID- 17427815 TI - Mirl is highly upregulated and localized to nuclei during infectious hyphal growth in the rice blast fungus. AB - Rice blast, caused by Magnaporthe grisea, is a devastating disease of rice throughout the world. Many recent molecular studies have focused on the early infection stages, but our knowledge about molecular events at the infectious hyphae stage is limited. In this study, 750 hygromycin-resistant transformants were isolated by transforming M. grisea Guyll with a promoterless enhanced green fluorescent protein (EGFP) construct. In one of the transformants, L1320, EGFP signals were observed in the nuclei of infectious hyphae. The transforming vector was inserted in a predicted gene named MIR1 and resulted in a Mir1 1-107-EGFP fusion. Mir1 is a low-complexity protein with no known protein domain and has no homolog in GenBank or other sequenced fungal genomes. Quantitative real-time reverse-transcriptase polymerase chain reaction analysis and expression assays of MIR1-EGFP fusion constructs indicated that the expression of MIR1 was highly induced during plant infection. Deletion analyses identified a 458-bp region that was sufficient for the MIR1 promoter activity. Further characterization revealed that a 96-bp sequence was essential for the enhanced in planta expression. MIR1 is an M. grisea-specific gene that is highly conserved among the field isolates belonging to the M. grisea species complex. The mir1 mutants had no obvious defects in appressorial penetration and rice infection. When overexpressed with the RP27 promoter, nuclear localization of the Mir1-EGFP fusion was observed in conidia and vegetative hyphae. These data suggest that the expression but not the nuclear localization of MIR1 is specific to infectious hyphae and that reporter genes based on MIR1 may be suitable for monitoring infectious growth in M. grisea. PMID- 17427816 TI - Heterochromatin-like regions as ecological niches for avirulence genes in the Leptosphaeria maculans genome: map-based cloning of AvrLm6. AB - Map-based cloning of avirulence genes of the AvrLml-2-6 cluster was recently undertaken in Leptosphaeria maculans and led to the identification of AvrLm1. The ensuing chromosome walk toward AvrLm6 resulted in the delineation of a 562-kb bacterial artificial chromosome (BAC) clone contig in an avirulent isolate. Following sequencing of the contig and sequence comparison with a virulent isolate, four AvrLm6 candidate genes were identified. Complementation of the virulent isolate with the four candidates was performed and one gene was found to fully restore the avirulent phenotype on Rlm6 oilseed rape genotypes. AvrLm6 was found to be located in the same genome context as AvrLml, because it is a solo gene surrounded by 85 and 48 kb of degenerated repeats on its 5' and 3' sides, respectively. AvrLm6 is an orphan gene encoding a small, potentially secreted, cysteine-rich protein. Comparison of AvrLm1 and AvrLm6 expressions by quantitative reverse-transcription polymerase chain reaction revealed that both genes are highly overexpressed during primary leaf infection. Using RNA interference, decreasing expression of AvrLm6 was shown to result in virulence toward Rlm6 genotypes whenever the expression was reduced by more than 60% compared with the wild-type isolate. PMID- 17427817 TI - The effect of chlorhexidine on dentin hybrid layers in vivo. AB - This in vivo study evaluated by TEM the degradation of dentin hybrid layers in deep occlusal resin composite restorations. Caries-free premolars scheduled for extraction as part of orthodontic treatment were prepared, restored and evaluated after two and six months. The adhesive used was a single-bottle etch-and-rinse product (Single Bond Plus, 3M ESPE). Control group restorations were placed according to the manufacturer's instructions, while the experimental group received application of a 2% solution of chlorhexidine digluconate after etching. No degradation was observed in either group after two months. Slight degradation was found in the control group after six months, but none was observed in the experimental group. In vitro testing showed no significant difference in microtensile bond strength between the control and experimental adhesive protocols. PMID- 17427818 TI - Postoperative sensitivity: a comparison of two bonding agents. AB - Historically, postoperative pain associated with temperature was considered a thermal conduction problem. More recently, pulpal hydrodynamics has been used to explain this sensitivity. Relative to restorations placed with dentin bonding agents that require a separate etching step, agents that include an acidic primer are believed to result in a better seal of the dentinal tubules. This study compared pain associated with a standardized cold stimulus in two groups of restorations. One group was placed with a self-priming resin that required a separate etch step, the other with a self-etching, self-priming dentin bonding agent. This was a community-based, randomized, double-blind clinical trial. Two hundred and nine restorations were placed for 76 participants. All teeth were asymptomatic at the start of the trial. Immediately following application of a standardized cold stimulus, participants rated the pain for each restored tooth using a Visual Analog Scale (VAS). For each group of restorations, VAS scores at 13 weeks were compared to preoperative scores. In addition, the preoperative score was subtracted from the 13-week score, and the two groups of restorations were compared. For both groups of restorations, the median scores were significantly reduced at 13 weeks. This decrease in the VAS score reflects a reduction in sensitivity below that which existed preoperatively. There was no significant difference between the two groups of restorations in terms of change in sensitivity at 13 weeks. PMID- 17427819 TI - Clinical performance and wear resistance of two compomers in posterior occlusal restorations of permanent teeth: six-year follow-up. AB - This study evaluated the clinical performance and wear resistance of compomer restorations placed in the occlusal cavities of posterior permanent teeth after six years. In 1999, 72 Class I restorations were placed by a single operator in 33 patients. Eighty-two percent of these restorations were located in molars. Each patient received at least two restorations, one with F2000 (3M ESPE) and another with Dyract AP (Dentsply). The finished and polished restorations that were free of any failure were considered the baseline. The restorations were clinically evaluated at baseline and at one-, two- and six-year intervals using modified USPHS criteria for color mismatch, marginal discoloration, surface roughness, marginal adaptation, anatomic form and secondary caries. Polyvinylsiloxane impressions (Express, 3M ESPE) were also taken, and models were obtained for indirect wear assessment (Leinfelder scale) at the same intervals. After six years, 11 patients attended the recall. Twenty-seven compomer restorations (11 with Dyract and 16 with F2000) were reevaluated. Data were submitted to the Friedman's test, ANOVA with repetitive measures, Tukey's test (clinical data), Wilcoxon and Kruskal-Wallis tests and the Spearman's correlation test (wear evaluation), all at a significance level of p < 0.05. When comparing the materials, F2000 and Dyract presented similar clinical performance and occlusal wear at the end of the clinical trial. The two compomers showed a significant increase in wear at the six-year follow-up, and a positive correlation (r2 = 0.65) was detected between wear and evaluation time (p < 0.001). Despite the decrease in restoration quality and the increase in occlusal wear, nearly all restorations were considered acceptable after the six-year evaluation. PMID- 17427820 TI - The effect of energy application sequence on the microtensile bond strength of different C-factor cavity preparations. AB - This study investigated the effect of three equivalent radiant exposure energy application sequences (EASs): high intensity power (HIP: 1,177 mW/cm2, 40 seconds), low intensity power (LOP: 573 mW/cm2, 71 seconds) and ramp (RMP: 650 mW/cm2, 5 seconds, then the irradiance increases to 1047 mW/cm2, 37 seconds) on the microtensile bond strength (MTBS) in both low and high C-factor cavity preparations. Thirty Paradigm blocks (Z100) were mounted in stone, with their top surfaces parallel to the mounting block. High C-factor (HC = 3.8) Class I cavity preparations were then prepared in 15 of the Paradigm blocks using a computer controlled specimen former. Low C-factor (LC = 1.8) Class II cavity preparations were prepared in the remaining 15 blocks by sectioning them perpendicularly using a slow-speed diamond saw. Five samples, one from each experimental group (3 EASs x 2 C-factors), were prepared and stored in the dark for 48 hours in distilled water at 37 degrees C prior to MTBS testing on the third day and on the remainder of the samples (n = 30). Mode of fracture was determined with a stereomicroscope at 20-40x magnification. The findings of this study indicated that HC, in combination with HIP, had significantly lower bond strength (27.54 MPa) than LC with HIP (51.39 MPa). On the other hand, there was no significant difference between high and low C-factors with the other EASs (LOP or RMP). There was also no significant effect for EAS (HIP, RAMP and LOP), with C-factor (HC or LC) held constant. The HIP-HC group had the highest percentage of adhesive (20%) and mixed adhesive (50%) failures (70% total). Adhesive and mixed adhesive failures occurred in other groups, but with lower percentages (RMP-LC: 40% total) (LOP-HC: 40% total). PMID- 17427821 TI - Increases in dentin-bond strength if doubling application time of an acetone containing one-step adhesive. AB - This study investigated the microtensile bond strength (microTBS) of a one-step self-etching adhesive to human dentin and bovine enamel following different bonding treatments. Occlusal portions of human molars and labial surfaces of bovine incisors were ground flat to provide uniform dentin and enamel surfaces, respectively. Futurabond was used following five different protocols: 1) according to the manufacturer's directions, 2) acid etched with 36% phosphoric acid (H3PO4) for 15 seconds, 3) 10% sodium hypochlorite (NaOCl) treated for two minutes after H3PO4-etching, 4) doubling the application time of the adhesive and 5) doubling the number of adhesive coats. Composite build-ups (6 mm in height) were constructed incrementally with Arabesk resin composite. The specimens were stored in 100% humidity for 24 hours at 37 degrees C and sectioned into beams of 1.0 mm2 cross-sectional area. Each beam was tested in tension in an Instron machine at 0.5 mm/minute, and mean microTBS data (MPa) were analyzed by one-way ANOVA and post-hoc multiple comparisons tests (alpha = 0.05). Doubling the application time of Futurabond attained the highest microTBS to dentin; whereas, no differences among all bonding application parameters evaluated could be detected when the adhesive was applied to enamel. PMID- 17427822 TI - Effects of temperature on the fluoride release and recharging ability of glass ionomers. AB - This study investigated the effects of environmental temperature on the fluoride release and recharging ability of glass ionomers. Five disk specimens (15 mm in diameter and 1 mm thick) were made of each of the following materials: a conventional luting glass ionomer, two high viscosity restorative glass ionomers and a restorative resin-modified glass ionomer. The fluoride release of each material was measured at 4 degrees C, 37 degrees C and 55 degrees C. An additional three groups, which were made of the same materials, were stored in distilled and deionized water for 30 days and recharged in 250 ppm fluoride solution at 4 degrees C, 37 degrees C and 55 degrees C for five minutes. The fluoride re-release was measured daily from two days prior to two days after the recharging process. At all temperatures, the luting glass ionomers showed the greatest fluoride release and recharging ability, followed by the resin-modified glass ionomer, then the high viscosity glass ionomers. For each material, the fluoride release increased with increasing temperature and all glass ionomers showed greater recharging ability at higher temperatures. An increase in environmental temperature increased both the fluoride release and recharging of the glass ionomers. This may be important in developing regimes for improving the delivery of topical fluoride products. PMID- 17427823 TI - Influence of curing rate of resin composite on the bond strength to dentin. AB - This study determined whether the strength with which resin composite bonds to dentin is influenced by variations in the curing rate of resin composites. Resin composites were bonded to the dentin of extracted human molars. Adhesive (AdheSE, Ivoclar Vivadent) was applied and cured (10 seconds @ 1000 mW/cm2) for all groups. A split Teflon mold was clamped to the treated dentin surface and filled with resin composite. The rate of cure was varied, using one of four LED-curing units of different power densities. The rate of cure was also varied using the continuous or pulse-delay mode. In continuous curing mode, in order to give an energy density totaling 16 J/cm2, the power densities (1000, 720, 550, 200 mW/cm2) emitted by the various curing units were compensated for by the light curing period (16, 22, 29 or 80 seconds). In the pulse-delay curing mode, two seconds of light curing at one of the four power densities was followed by a one minute interval, after which light cure was completed (14, 29, 27 or 78 seconds), likewise, giving a total energy density of 16 J/cm2. The specimens produced for each of the eight curing protocols and two resin composites (Tetric EvoCeram, Ivoclar Vivadent; Filtek Supreme XT, 3M ESPE) were stored in water at 37 degrees C for seven days. The specimens were then either immediately subjected to shear bond strength testing or subjected to artificial aging (6,000 cycles between 5 degrees C and 55 degrees C baths) prior to testing. Failure modes were also assessed. The shear bond strengths were submitted to factorial analysis of variance, and the failure modes were submitted to a Chi-square test (alpha = 0.05). All but power density (curing mode, resin composite material and mode of aging) significantly affected shear bond strength. The curing mode and resin composite material also influenced the failure mode. At the selected constant energy density, pulse-delay curing reduced bonding of the resin composite to dentin. PMID- 17427824 TI - Shear bond strength of dual-cured and self-cured resin composites to dentin using different bonding agents and techniques. AB - This study determined the effects of bonding agents on the shear bond strength of dual- and self-cured resin composites to dentin. Two light-cured dentin bonding agents (Excite and One-Step) and a dual-cured bonding agent (Excite DSC) were compared. Light activation of the bonding agents prior to placement of the resin composites was also evaluated. This in vitro study was performed on 120 extracted non-carious human third molars. The occlusal part of the crowns was removed to expose a flat dentin surface. The teeth were then randomly divided into three major groups for Excite, One-Step and Excite DSC as bonding agents. The specimens in each adhesive group were divided into four subgroups: with and without light activation of the bonding agent and with dual-cured (Luxacore Dualcure, DMG, Hamburg, Germany) or light-cured resin (Luxacore, DMG, Hamburg, Germany) composites. After placing the restorations, the specimens were kept in water at 37 degrees C for 24 hours before being tested for shear bond strength on an Instron universal testing machine at a crosshead speed of 0.5 mm/minute. The results showed that the shear bond strength of dual-cured resin composite to dentin was significantly higher than that of self-cured resin composite (p = 0.017). Light activation of the bonding agents prior to applying the resin composites led to a significantly higher shear bond strength of the resin composites to dentin, compared to no light activation (p < 0.05). PMID- 17427825 TI - The effect of soft-start polymerization by second generation LEDs on the degree of conversion of resin composite. AB - Fourier-Transform (FT)--Raman spectroscopy was used to evaluate in vitro the degree of conversion (DC) of Charisma dental composite cured by three different light curing units (LCUs) using soft-start and normal protocols. Eighty circular blocks of resin (7 mm in diameter x 2.5 mm thick) were prepared and cured using the following sources: halogen light (Degulux soft-start, n = 20, G1-G2), low power light emitting diode (LED) with transparent polymer tip (LD13, n = 20, G3 G4) and fiber optic tip (LD13, n = 20, G5-G6) and, finally, high power LED (Radii, n = 20, G7-G8). The top and bottom surfaces of the blocks were analyzed by FT-Raman spectroscopy. The respective DCs were estimated calculating the peak height ratio of the aliphatic C=C (1640 cm(-1)) and aromatic C=C (1610 cm(-1)) Raman modes. The DC at the surfaces ranged between 50% and 60% for the top and 46% and 58% for the bottom. The halogen light and high power LED LCUs with the soft-start and normal protocols produced the highest DC values of dental composite at both surfaces (p < 0.001). Curing by low power LED in the soft-start protocol did not produce adequate DC at the depth of 2.5 mm. The type of LCU light guide tip did not present a significant statistical difference in the final DC of the dental composite (p > 0.05). PMID- 17427826 TI - Bond strength of different adhesive systems to dental hard tissues. AB - This study compared the shear bond strengths of different adhesive systems to enamel and dentin of different depths. The adhesive systems used were: Single Bond one-bottle total-etch; AQ Bond one-step self-etching, Clearfil SE Bond two step self-etching and Tyrian SPE/One-step Plus two-step self-etching. Eighty extracted non-carious human mandibular molars were mounted in self-curing resin and the occlusal surfaces were ground with a mechanical grinder to obtain flat occlusal enamel surfaces. After applying the adhesive systems, a plastic tube was attached to the enamel surfaces. The tube was filled with a universal hybrid resin composite, which was then polymerized. The specimens were stored in water at 37 degrees C for 24 hours. Shear bond testing was carried out using an Instron Universal testing machine with a crosshead speed of 1 mm/minute. The occlusal enamel of the 80 teeth was removed to determine the bond strengths of the adhesives to superficial dentin. To obtain deep dentin, the same teeth were ground deep towards the pulp, with the remaining dentin thickness approximately 0.7 mm. The adhesives and restorative material were then applied to the prepared dentin surfaces following the methodology described above. For occlusal enamel surfaces, the bond strength of Single Bond was significantly higher than the other adhesives. Single Bond, Clearfil SE Bond and Tyrian SPE/One-step Plus performed equally when bonded to superficial dentin; the lowest bond strength was obtained with AQ Bond. On deep dentin, the highest bond strengths were obtained with Clearfil SE Bond. For each adhesive system, shear bond strength to enamel was statistically higher than it was to either superficial or deep dentin. There were no significant differences between shear bond strengths to superficial versus deep dentin, regardless of adhesive. PMID- 17427827 TI - Bond strengths of all-ceramics: acid vs laser etching. AB - Various applications of dental lasers on dental materials have been proposed for surface modifications. This study evaluated whether laser etching could be an alternative to hydrofluoric acid (HF) etching. One hundred and ten lithia-based all-ceramic specimens (Empress 2) (R: 4 mm, h: 4 mm) were prepared and divided into five groups (n = 22/group). The untreated specimens served as the control, while one of the experimental groups was treated with 9.5% HF for 30 seconds. Three remaining test groups were treated with different laser (Er:YAG laser wavelength:2940 nm, OpusDent) power settings: 300 mJ, 600 mJ and 900 mJ. Ten specimens in each group were luted to the other 10 specimens by a dual-curing cement (Variolink II), and shear-bond strength (SBS) tests were performed (Autograph, crosshead speed: 0.5 mm/minute). The results were statistically analyzed (Kruskal Wallis and Mann Whitney-U, alpha = .05). Mean SBS (MPa) were 31.9 +/- 4.0, 41.4 +/- 4.3, 42.8 +/- 6.2, 29.2 +/- 4.5 and 27.4 +/- 3.8 for the control and HF, 300, 600 and 900 mJ groups, respectively. SEM evaluations revealed different surface morphologies depending on the laser parameters. The differences between HF acid and 300 mJ, when compared with the control, 600 and 900 mJ groups, were significant (p < .05). The 300 mJ laser group exhibited the highest shear-bond strength values, indicating that laser etching could also be used for surface treatments. PMID- 17427828 TI - Restoration interface microleakage using one total-etch and three self-etch adhesives. AB - This study evaluated the efficacy of a total-etch and three self-etch adhesives in reducing microleakage after three months water storage and thermocycling. Thirty freshly extracted caries-free human premolars and molars were used. Class V standardized preparations were performed on the facial and lingual surfaces, with the gingival margin placed 1 mm below the CEJ. The teeth were randomly divided into four groups; Group I: Xeno III one-step self-etch adhesive (Dentsply/Caulk), Group II: Prime & Bond NT total-etch adhesive (Dentsply/Caulk), Group III: i-Bond one-step self-etch adhesive (Heraeus Kulzer) and Group IV: Clearfil SE Bond two-step self-etch adhesive (Kuraray Medical). The teeth were restored using 2 mm increments of shade A2 resin composite (Esthet-X, Dentsply/Caulk). Each layer was cured using the Spectrum 800 curing light (Dentsply/Caulk) for 20 seconds at 600mW/cm2. The teeth were stored in distilled water for 90 days. Samples were thermocycled 500x between 5 degrees C and 55 degrees C with a dwell time of 30 seconds, then placed in a 0.5% methylene blue dye solution for 24 hours at 37 degrees C. Samples were sectioned longitudinally and evaluated for microleakage at the occlusal and gingival margins under a stereomicroscope at 20x magnification. Dye penetration was scored: 0 = no penetration; 1 = partial dye penetration along the occlusal or gingival wall; 2 = dye penetration along the occlusal or gingival wall; 3 = dye penetration to and along the axial wall. A Mann-Whitney test was used to demonstrate significantly more dye penetration in Group III than in the other groups at both the occlusal and gingival scores (p < 0.0001). When comparing the occlusal and gingival scores for each group, the Wilcoxon Rank test showed no significant difference in dye penetration for Xeno III (p > 0.05), Prime & Bond NT (p = 0.059) and I Bond (p = 0.083), and Clearfil SE Bond yielded more dye penetration at the occlusal than at the gingival wall (p = 0.001). PMID- 17427829 TI - Surface roughness of novel resin composites polished with one-step systems. AB - OBJECTIVES: This study: 1) analyzed the surface roughness of five novel resin composites that contain nanoparticles after polishing with three different one step systems and 2) evaluated the effectiveness of these polishers and their possible surface damage using scanning electron microscope (SEM) analysis. METHODS: The resin composites evaluated in this study include CeramX, Filtek Supreme XT, Grandio, Premise and Tetric EvoCeram. A total of 100 discs (20/resin composites, 10 x 2 mm) were fabricated. Five specimens/resin composites cured under Mylar strips served as the control. The samples were polished for 30 seconds with PoGo, OptraPol and One Gloss discs at 15,000 rpm using a slow speed handpiece. The surfaces were tested for roughness (Ra) with a surface roughness tester and examined with SEM. One-way ANOVA was used for statistical analysis (p = 0.05). RESULTS: For all the composites tested, differences between the polishing systems were found to be significant (p < 0.05). For Filtek Supreme XT, Mylar and PoGo created equally smooth surfaces, while significantly rougher surfaces were obtained after OptraPol and One Gloss applications. For Grandio, Mylar and PoGo created equally smooth surfaces, while OptraPol and One Gloss produced equally rougher surfaces. Tetric EvoCeram exhibited the roughest surface with OptraPol, while no significant differences were found between Premise and CeramX. According to SEM images, OptraPol and One Gloss scratched and plucked the particles away from the surface, while PoGo created a uniform finish, although the roughness values were not the same for each composite. CONCLUSION: Effectiveness of the polishers seems to be material dependent. PMID- 17427830 TI - A case for rubber dam application--subcutaneous emphysema after Class V procedure. PMID- 17427831 TI - Academy of Operative Dentistry Hollenback Memorial Prize. Professor Dr Guido Vanherle. PMID- 17427832 TI - Academy of Operative Dentistry Award of Excellence. Dr Thomas G Berry. PMID- 17427833 TI - Prevalence of isolated systolic hypertension in the population over 55 years old. Results from a national study. AB - INTRODUCTION: Isolated systolic hypertension (ISH) is an important risk factor for cardiovascular events (myocardial infarction and stroke) and premature mortality, particularly in the elderly population. Among this population, it is an even better predictor of morbidity and mortality than diastolic blood pressure (BP). This cooperative survey was promoted by the World Hypertension League, Associacao Portuguesa de Hipertensao, Instituto Nacional de Cardiologia Preventiva, Sociedade Portuguesa de Cardiologia, Sociedade Portuguesa de Geriatria, Sociedade Portuguesa de Medicina Intema, and Direccao Geral de Saude, with the collaboration of Servier Portugal. OBJECTIVES: The aim of this survey was to evaluate the prevalence of ISH within the Portuguese elderly population (aged 55+) during the year 2000, compared and together with other countries, in order to estimate its worldwide prevalence. Secondary objectives were to investigate demographic factors related to ISH (age, gender and other cardiovascular risk factors) and to analyze the effects of repeated blood pressure measurements on prevalence and on postural variations in blood pressure. METHODS: We performed a survey including 108 primary care centers. General practitioners were invited to include the first consecutive 15 patients aged over 55 years. Subjects were be selected over a period of one week and had to agree to attend three visits. All blood pressure measurements were to be performed in accordance with the standards of the 7th Report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure (JNC VI). 1031 subjects were included in this survey. CONCLUSIONS: We concluded that ISH prevalence ranges between 20.3% in primary" care patients and 35% in the community, using mean values of both systolic and diastolic BP. ISH prevalence is higher among women; it increases with age, decreasing slightly after the age of 70. Risk factors include gender, age and family history of hypertension. The predictive value for ISH diagnosis is 100% and 65% with methods ISH-1 and ISH-2 respectively, with positive predictive value of 51.05% and 63% for ISH-1 and ISH 2 respectively. Orthostatic hypotension prevalence is higher at 5 minutes than at 2 minutes and is higher among elderly subjects. The prevalence of untreated hypertension in the Portuguese population aged over 55 years, according to the WHO/ISH criteria, was 44.92%. PMID- 17427834 TI - Prevalence, awareness, treatment and control of hypertension in Portugal. The PAP study. AB - The objective of this study was to estimate the prevalence and distribution of hypertension and to determine the level of awareness, treatment, and control of hypertension in the Portuguese adult population. The study was conducted in 2003, using a multistage cluster sampling method to select a representative national sample. A total of 5023 adults, aged 18 to 90 years, were examined. Three blood pressure measurements were obtained by trained observers using an OMRON M4-1 sphygmomanometer after a 5-minute seated rest. Information on history of hypertension and use of antihypertensive medication was obtained using a standard questionnaire. Hypertension was defined as mean systolic blood pressure > or =140 mmHg and/or diastolic blood pressure > or =90 mmHg, or use of antihypertensive medication. Overall, 42.1% of the Portuguese adult population aged 18 to 90 years, representing 3,311,830 people, would have hypertension. The age-specific prevalence of hypertension in the three age-groups studied--under 35 years, 35-64 years, and over 64 years--was 26.2%, 54.7% and 79% in men and 12.4%, 41.1% and 78.7% in women respectively. Among hypertensive subjects, only 46.1% were aware of their high blood pressure, 39.0% were taking antihypertensive medication, and 11.2% had their blood pressure controlled (<140/90 mmHg). Our results indicate that hypertension is highly prevalent in Portugal. The percentages of hypertensives who are aware of their condition, are being treated, and whose hypertension is controlled are unacceptably low. These results underscore the urgent need to develop national strategies to improve prevention, detection, and treatment of hypertension in Portugal. PMID- 17427835 TI - Isolated pulmonary valve endocarditis due to Pseudomonas aeruginosa. AB - Isolated endocarditis of the native pulmonary valve is a rare clinical condition. It usually appears in association with certain predisposing factors, particularly intravenous drug abuse, alcohol abuse, sepsis, endovascular infections or congenital heart disease. The authors describe the case of isolated pulmonary valve endocarditis due to Pseudomonas aeruginosa of possible nosocomial origin in the absence of the predisposing factors mentioned above. Native pulmonary valve infection due to Pseudomnzonas aeruginosa in the absence of predisposing factors has not been described in the literature. The authors stress the importance of diagnosing this entity as early as possible as it is associated with high mortality and is on the increase due to the growing number of invasive procedures applied to patients in the hospital environment. PMID- 17427836 TI - The role of statins in aortic stenosis. Myth or reality? AB - Our understanding of aortic valve disease has improved in the past decade from considering it a degenerative process to the realization that it is an active biologic disease. Aortic valve disease, although it has a similar atherosclerotic pathogenesis to vascular disease, differs in terms of bone calcification. Determining the appropriate timing of statin therapy to slow the progression of bone formation in these lesions is essential for the routine use of these drugs in such patients. Knowledge of the biology of valve lesions will play an important part in understanding this disease and future treatment options for these patients. PMID- 17427837 TI - Congenital pericardial cyst. PMID- 17427838 TI - Left ventral apical pericardial cyst. PMID- 17427839 TI - National Registry on Cardiac Electrophysiology--2005. PMID- 17427840 TI - Medicaid information technology architecture: an overview. AB - The Medicaid Information Technology Architecture (MITA) is a roadmap and tool-kit for States to transform their Medicaid Management Information System (MMIS) into an enterprise-wide, beneficiary-centric system. MITA will enable State Medicaid agencies to align their information technology (IT) opportunities with their evolving business needs. It also addresses long-standing issues of interoperability, adaptability, and data sharing, including clinical data, across organizational boundaries by creating models based on nationally accepted technical standards. Perhaps most significantly, MITA allows State Medicaid Programs to actively participate in the DHHS Secretary's vision of a transparent health care market that utilizes electronic health records (EHRs), ePrescribing and personal health records (PHRs). PMID- 17427841 TI - Clinical health information technologies and the role of medicaid. AB - Clinical health information technologies (HIT) are widely viewed as essential tools for improving the quality and efficiency of health care delivery. Medicaid agencies make substantial investments in information technology (IT), have much to gain through the widespread use of clinical HIT, and can have significant influence on the adoption of HIT by providers. Medicaid agencies, however, face legal, regulatory, and financing challenges in relation to supporting HIT adoption, use, standardization, and interoperability. This article summarizes the issues related to Medicaid's participation and support of clinical HIT, and makes recommendations for addressing policy challenges at the State and Federal level. PMID- 17427842 TI - Medicaid and health information: current and emerging legal issues. AB - Legal questions are an inevitable byproduct ofsignificant technology change in health care such as that underway as a result of health information technology (HIT). This article examines several important existing and emerging legal questions in a Medicaid context. First, do the Centers for Medicare & Medicaid Services (CMS) and State Medicaid agencies, have a fiduciary obligation to adopt and fully use health information technology given its potential to improve health care quality while reducing racial, ethnic, and socioeconomic disparities in health and health care? Second, how can Medicaid privacy standards be reconciled with the Health Insurance Portability and Accountability Act (HIPAA) privacy rule? Third, what actual or perceived legal barriers exist to ensuring that Medicaid information is interoperable with data produced under critical health care, educational, and social programs from which beneficiaries are simultaneously receiving care? PMID- 17427843 TI - Valuing hospital investment in information technology: does governance make a difference? AB - This article examines the investment of patient care information technology (IT) systems by a nationwide sample of U.S. short-term acute care hospitals and the resulting impact these systems have in the productivity of institutions from 1990 1998. Of particular interest is the extent to which for-profit and not-for-profit hospitals obtain different results from the adoption of lT systems. We find that the marginal effect of IT on for-profit hospital productivity is to reduce the number of days supplied, while in not-for-profit hospitals the marginal effect of IT is to increase the quantity of services supplied. This resulting effect is consistent with the differing objectives of not-for-profit and for-profit hospitals and demonstrates the positive marginal value of IT as a sustainable and prudent investment. PMID- 17427844 TI - Medicare beneficiaries' use of computers and internet: 1998-2005. PMID- 17427845 TI - Role of SCHIP in serving children with special health care needs. AB - The purpose of this article is to provide new information about two policy issues: (1) Is the State Children's Health Insurance Program (SCHIP) an important source of health insurance for children with special health care needs (CSHCN)? and (2) Does SCHIP provide CSHCN with better access to care, compared with other insurance coverage? Using the 2001 National Survey of CSHCN, we found that a limited fraction of CSHCN were eligible for SCHIP while a relatively small proportion of SCHIP-eligible CSHCN were uninsured. Access to care for CSHCN under SCHIP was better than those SCHIP-eligible but uninsured, and similar to those income-eligible for SCHIP but privately insured. PMID- 17427846 TI - End-of-life expenditures by Ohio Medicaid beneficiaries dying of cancer. AB - We evaluate the extent to which the Ohio Medicaid Program serves as a safety net to terminally ill cancer patients, and the costs associated with providing care to this patient population. Over a 10-year period, Ohio Medicaid served nearly 45,000 beneficiaries dying of cancer, and spent more than $1 billion in medical care expenditures in their last year of life. Eighty percent of the expenditures were incurred by 67 percent of the decedents who had been enrolled in Medicaid for at least 1 year before death, implying an opportunity for the Medicaid Program to ensure timely transition to palliative care and hospice. PMID- 17427847 TI - Trends in the health status of medicare risk contract enrollees. AB - Previous research has found Medicare risk contract enrollees to be healthier than beneficiaries in fee-for-service (FFS). Medicare Current Beneficiary Survey (MCBS) data were used to examine trends in health and functional status measures among risk contract and FFS enrollees from 1991 to 2004. Risk contract enrollees reported better health and functioning, but the differences tended to narrow over time. Most of the differences in trends were observed for functional status measures and institutionalization; differences in trends for perceived health status and prevalence rates of chronic conditions tended to be small or non existent. The narrowing of functional and health status differences between the risk contract and FFS populations may have implications for payment policy, as well as implications for the role of private health plans in Medicare. PMID- 17427848 TI - Setting physicians' prices in FFS medicare: an economic perspective. AB - Recent policy discussions by the Medicare Payment Advisory Commission (MedPAC) regarding physician prices in the traditional fee-for-service (FFS) Medicare Program reflect movement toward a market pricing model. Earlier objectives such as sustainable levels of spending have given way to concerns over the relationship between fees and actual costs, access to care, and the importance of demand and supply in local markets. An important objective in other policy settings is economically efficient distribution of services. We explain the meaning of economic efficiency for Medicare physician prices and explore difficulties one might encounter in pursuing economic efficiency, as well as the cost of not pursuing it. PMID- 17427849 TI - [Contribution to healing]. PMID- 17427850 TI - [From the Salvation Army to the Mormons (interview by Irene Neubauer)]. PMID- 17427851 TI - [Like a feather drifting away]. PMID- 17427852 TI - [Sexual rehabilitation of heart patients]. PMID- 17427853 TI - [Shoes as a safety risk]. PMID- 17427854 TI - [Creating dignity anew in every process]. PMID- 17427855 TI - [Tolerating the tension of feelings (1.)]. PMID- 17427856 TI - [Preventing limitations as much as possible]. PMID- 17427857 TI - [Discovering or rediscovering collaboration]. PMID- 17427858 TI - [The image the nursing profession has of itself]. PMID- 17427859 TI - [Applying the policy to practice]. PMID- 17427860 TI - [Activity of trainees under the magnifying glass]. PMID- 17427861 TI - ["Before each new step, taking time to reflect]. PMID- 17427862 TI - [A question of priority]. PMID- 17427863 TI - The importance of CE in tough times. PMID- 17427864 TI - The effect of a patient's own negligence. PMID- 17427865 TI - Reference checks: what to say when you get the call. PMID- 17427866 TI - How to get the most out of your MDA Annual Session. PMID- 17427867 TI - Life insurance basics you should know. PMID- 17427868 TI - Moderate coffee drinking may have many benefits. PMID- 17427869 TI - Dr. Arthur Victor: America's oldest practicing dentist. PMID- 17427871 TI - Successful associateships don't just "happen". PMID- 17427870 TI - Keeping up with the latest in regulatory compliance. PMID- 17427872 TI - Functional specialisation within the cortical language network: effects of cortical dysfunction. AB - In the 1990's neuroanatomical models of language and semantic memory have been mainly based on functional neuroimaging studies of brain activity in healthy volunteers and correlational studies between structural lesions in patients and behavioral deficits. In this paper we present a novel approach where we test models that have been developed in healthy volunteers by means of functional imaging in patients in combination with behavioral studies. Study populations consist of patients with focal cortical stroke (n = 2), amnestic mild cognitive impairment (n = 14) and primary progressive aphasia (n = 18). The experiments provide converging evidence that 1. the integrity of the right mid- and anterior fusiform gyrus is required for full and detailed retrieval of knowledge of visual attributes of concrete entities 2. the left posterior superior temporal sulcus is critically involved in lexical-semantic retrieval 3. the anterior temporal pole to the left functions as an associative structure that links the representations of meaning that are distribured over the cortical brain surface. Our experiments also provide us with new insight into the degradation and re-organisation of the language system in cortical neurodegenerative disease. PMID- 17427873 TI - Gene expression profiling of primary cutaneous melanoma. AB - Cutaneous Malignant Melanoma (CMM) is the most malignant skin tumour in humans, the incidence of which is rising rapidly in most fair-skinned populations, without apparent decline in mortality. Both hereditary, constitutional and environmental factors play a role in its etiology. CMM arises from melanocytes in the epidermis, and proceeds through discrete steps of tumor-progression that consist histologically of the radial growth phase (RGP), vertical growth phase (VGP) and metastatic phase. The underlying molecular mechanisms that govern the transition between these growth phases are hardly known. The prognosis of patients with VGP melanoma depends on several clinical and histological parameters; the latter include thickness, mitotic activity, presence or absence of ulceration and regression, and pattern of lymphocytic host response. However, there is still need for new prognostic parameters. To obtain insight in the molecular mechanisms of tumor progression in CMM, and in search of new prognostic markers, we performed global gene-expression profiling using 44K oligonucleotide micro-arrays on a unique retrospective series of 83 frozen primary MM with VGP, 9 metastases and 23 benign nevi. Unsupervised analysis allowed us o identify clusters of melanoma patients with different outcome based on their gene expression profile only. Supervised analysis resulted in the identification of a genomic signature of 254 genes with prognostic significance. The large majority of the 254 enes was correlated with thickness, thereby stressing the importance of thickness in he prognosis of CMM. This signature was validated on a separate series of melanoma patients, and proved to have a predictive accuracy comparable to what can be obtained by tumour thickness and ulceration. On an immunohistochemical level, we identified 8 new markers that may help in the prognostication of melanoma patients; three of these markers, i.e. the mini chromosome maintenance (mcm) proteins mcm3, 4 and 6, hat are involved in DNA replication, had independent prognostic value. Additionally, upervised analysis showed similarities in gene expression profile between primary CMM and their metastases. In conclusion, our data provide new information regarding the molecules that are operative in the progression of CMM. CMM is notorious for its resistance to chemotherapy, and disseminated CMM is a uniformly fatal disease. As several of the progression-related genes, encode molecules that have been the target of established or xperimental cancer therapies, our results may hopefully contribute to the treatment of end-stage CMM-patients. PMID- 17427874 TI - Acquired chromosomal rearrangements targeting selected transcription factors: contribution of molecular cytogenetic and expression analyses to the identification of clinically and biologically relevant subgroups in hematological malignancies. AB - In leukemias chromosomal aberrations, balanced translocations in particular, play a critical role in the oncogenic process. The characterization of these chromosomal alterations was crucial to the discovery of the genes implicated in leukemogenesis, as the chromosomal breakpoints indicated their genomic localization. In addition, these molecular defects may serve as targets for diagnostic essays and can have a major prognostic value. Finally, the characterization of the deregulated cellular pathways potentially identifies targets for therapeutic intervention. In this paper we summarize our efforts to expand the current knowledge of the diagnostic, prognostic or biological significance of selected chromosomal aberrations identified in M-FISH studies. First, we illustrated the power of M-FISH in dissecting complex chromosomal aberrations in myeloid neoplasms. MLL amplification was defined as a clinical entity characterized by adverse prognosis and within the multitude and variety of chromosomal rearrangements a pattern of a limited number of cytogenetic subclasses was discerned. In leukemias characterized by 11q23 amplification, we described the amplicon and confirmed MLL, in addition to DDX6, as a principal amplification target. Molecular characterization of a large series of unselected sporadic and recurrent 3q26 rearranged leukemias confirmed the decisive role of ectopic EVI1 expression in these malignancies. We contributed to an extensive analysis of the phenotypical and prognostic features of T-ALL characterized by HOX11L2 expression and identified HOX11L2 overexpression as one of the most frequent genetic defects in childhood T-ALL, associated with intermediate prognosis. Finally, we designed and validated diagnostic tools for the detection of the t(9;14) (p13;q34) resulting in PAX5 overexpression and convincingly associated the presence of this rearrangement to high-grade morphology and karyotype complexity. In conclusion, the series of investigations presented here clearly illustrate the benefits of M-FISH as molecular tool for the dissection and characterization of complex and cryptic rearrangements. The subsequent reports demonstrate the utility of molecular cytogenetics and expression analyses to the clinical management of patients diagnosed with hematological malignancies. PMID- 17427875 TI - Temperature rising. Feeling a bit warm? You may just have to live with it. PMID- 17427876 TI - Morbidity and mortality patterns in HIV-1 seropositive/ seronegative women in Kampala and Harare during pregnancy and in the subsequent two years. AB - OBJECTIVE: To compare birth outcomes, hospital admissions and mortality amongst HIV-1 seropositive and HIV-1 seronegative pregnant women in Kampala, Uganda and Harare, Zimbabwe. DESIGN: In Kampala and Harare about 400 HIV-1 seropositive and 400 HIV-1 seronegative pregnant women were recruited at initial visit for antenatal care into a prospective study and followed for two years after delivery. The women were classified as HIV-1 seropositive at recruitment if initial and second ELISA tests were positive and confirmed by Western Blot assay. Data on demographic, reproductive, contraceptive and medical histories were obtained using a comprehensive questionnaire at entry, 32 and 36 weeks gestation, at delivery and at six, 12, and 24 months post delivery. In addition, a physical examination and various blood tests were performed at each antenatal and post natal visit. RESULTS: During the two years after delivery, HIV-1 seropositive women had higher hospital admission and death rates than HIV-1 seronegative women. HIV-1 seropositive mothers had a two-fold increase in risk of being admitted to hospital (Kampala: RR = 2.09; 95% CI = 0.95 to 4.59; Harare: RR = 1.98; 95% CI = 1.13 to 3.45). In the six weeks after delivery eight deaths occurred, six of which were among HIV-1 seropositive women and in the period from six weeks to two years after delivery, 53 deaths occurred, 51 of which were among HIV-1 seropositive women (Kampala: RR = 17.7; 95% CI = 4.3 to 73.2; Harare: RR = 10.0; 95% CI = 2.3 to 43.1). However, there was no difference in hospital admission rates between HIV-1 seropositive and seronegative women during pregnancy itself and there was only one death during that period (in a HIV-1 seronegative woman). There was no difference in the frequency of complications of delivery between HIV-1 seropositive and HIV-1 seronegative women and the outcome of births were also similar. CONCLUSIONS: A significant number of HIV-1 positive pregnant women presented at both Harare and Kampala although there was no difference in the number of hospital admissions or mortality between HIV-1 seropositive and HIV-1 seronegative women during pregnancy. Although there were no differences in complications during pregnancy or outcome at delivery, in the two years after delivery, HIV-1 seropositive women in both centres were at increased risk of being admitted to hospital and of dying. PMID- 17427877 TI - Occurrence of diabetogenic changes in pregnancy among black women in an urban setting. AB - OBJECTIVE: To find out if pregnancy in black Zimbabwean women is a diabetogenic state using basal blood levels of cortisol, insulin, C-peptide and glucose. METHODS: 111 women (28 non-pregnant, 29 first trimester, 26 second trimester and 28 third trimester) aged between 18 and 35 years were recruited for the study. Fasting plasma cortisol, insulin, C-peptide and glucose were determined by standard methods. The glucose/insulin ratio was used as an index of insulin sensitivity and the C-peptide/glucose ratio as well as the homeostasis assessment model (HOMA) as an index of insulin resistance. RESULTS: The means of fasting plasma cortisol levels were significantly elevated, p < 0.0001 among the four groups (non-pregnant, first, second and third trimester women). Fasting plasma insulin levels peaked during the third trimester and significant differences were noted among all women, p < 0.05. Similar data was obtained for C-peptide levels (a better indicator of beta-cell insulin secretory activity) among the groups, p < 0.01. The means of fasting plasma glucose levels were significantly decreased with advancing gestation, p < 0.0001. Significantly lower glucose/insulin ratios, a measure of insulin sensitivity and elevated C-peptide/glucose ratios, an index of insulin resistance, were demonstrated among the women, (p < 0.05 and < 0.01 respectively). CONCLUSION: The basal data presented in this paper clearly demonstrates that the diabetogenic effects of pregnancy are also expressed by Zimbabwean black women, especially in late gestation. PMID- 17427878 TI - Kidney transplantation in a developing economy: challenges and initial report of three cases at Ile Ife. AB - BACKGROUND: Kidney transplantation (KT) is globally adjudged the best alternative treatment for end stage renal disease (ESRD) in preference to life-long dialysis. This form of therapy was hitherto unavailable in Nigeria until our hospital and a private hospital embarked on a KT programme despite our depressed economy, and inadequate facilities. We present the initial report of KT performed in our hospital and the challenges of KT in our developing society. CASE REPORTS: Three patients with ESRD had living related KT between June 2002 and April 2003. The first patient died with functioning graft six and a half months post transplantation from complications of Diabetes mellitus and sepsis, while the remaining two still enjoy a good quality of life 35 months post transplantation. There were problems with procurement and monitoring of immunosuppressive drugs in the three patients. This report also illustrates the common causes of ESRD in Nigeria and some of the complications of KT. To our knowledge, these are the first reported cases of KT in Nigeria. CONCLUSION: Kidney transplantation is cost effective and offers a good quality of life for ESRD patients. Poverty, inadequate facilities and lack of donors are major problems facing KT in our society. Although KT requires high technical and material resources, with proper training, commitment and adequate funding, it is feasible, safe and cheaper on a long term basis for the management of patients with ESRD in a developing economy like ours. There is a need for government funding of KT programmes in developing countries. PMID- 17427879 TI - Low output from health professionals training schools contributing to human resources crisis in Malawi. PMID- 17427880 TI - Transplant rules changing. CMS pushing toward outcome-focused system. PMID- 17427881 TI - Community breaks out. After years of quiet growth, Community Health makes risky play for Triad to become top publicly traded hospital company. AB - Community Health Systems has long been a quiet company, slowly building its portfolio with small acquisitions. With its bid for Triad, Community would become the largest publicly traded hospital company in the U.S. While the company isn't talking yet about possible divestitures, James Burgdorfer, left, says most similar large deals result in sell-offs. PMID- 17427882 TI - Cashing in at Triad. PMID- 17427883 TI - Plans aplenty... but universal coverage still seeking a foundation. PMID- 17427884 TI - A boost for transparency. PMID- 17427885 TI - Doc ownership battle looms. PMID- 17427886 TI - Weighing the cost of a pandemic. PMID- 17427887 TI - Medicaid under siege. While some aim at universal coverage, a safety net program keeps taking hits. PMID- 17427888 TI - Evidence of an evolution. Our annual construction survey indicates the building boom continues--with research-based design playing a key role. PMID- 17427889 TI - Supplying faith. GPOs seeking even stronger ties to Catholic hospitals, health systems. PMID- 17427890 TI - Magnetic-field-induced vertigo: a theoretical and experimental investigation. AB - Vertigo-like sensations or apparent perception of movement are reported by some subjects and operators in and around high field whole body magnetic resonance body scanners. Induced currents (which modulate the firing rate of the vestibular hair cell), magneto-hydrodynamics (MDH), and tissue magnetic susceptibility differences have all been proposed as possible mechanisms for this effect. In this article, we examine the theory underlying each of these mechanisms and explore resulting predictions. Experimental evidence is summarised in the following findings: 30% of subjects display a postural sway response at a field gradient product of 1 T(2)m(-1); a determining factor for experience of vertigo is the total unipolar integrated field change over a period greater than 1 s; the perception of dizziness is not necessarily related to a high value of the rate of change of magnetic field; eight of ten subjects reported sensations ranging from mild to severe when exposed to a magnetic field change of the order of 4.7 T in 1.9 s; no subjects reported any response when exposed to 50 ms pulses of dB/dt of 2 Ts(-1) amplitude. The experimental evidence supports the hypothesis that magnetic-field related vertigo results from both magnetic susceptibility differences between vestibular organs and surrounding fluid, and induced currents acting on the vestibular hair cells. Both mechanisms are consistent with theoretical predictions. PMID- 17427891 TI - A few remarks on 'combined action of DC and AC magnetic fields on ion motion in a macromolecule'. AB - Zhadin and Barnes [2005:26:323-330] concluded that they solved the differential equation describing combined action of DC and AC magnetic fields on thermal motion of ions in a biological macromolecule and, as a result, a diversity of biological phenomena could be explained. It is shown here that biological phenomena cannot be explained based on this model. Adair [2006:27:332-334] gave several arguments for the statement that the interaction of weak magnetic fields with ions trapped in protein cavities cannot produce detectable biological effects through changing the character of the ion orbits. The arguments are analyzed here and some are shown to be questionable or unjustified. We stress the difference between the conclusion made by Adair and that stated in this article. PMID- 17427893 TI - Catalytic carbophilic activation: catalysis by platinum and gold pi acids. AB - The ability of platinum and gold catalysts to effect powerful atom-economic transformations has led to a marked increase in their utilization. The quite remarkable correlation of their catalytic behavior with the available structural data, coordination chemistry, and organometallic reactivity patterns, including relativistic effects, allows the underlying principles of catalytic carbophilic activation by pi acids to be formulated. The spectrum of reactivity extends beyond their utility as catalytic and benign alternatives to conventional stoichiometric pi acids. The resulting reactivity profile allows this entire field of catalysis to be rationalized, and brings together the apparently disparate electrophilic metal carbene and nonclassical carbocation explanations. The advances in coupling, cycloisomerization, and structural reorganization--from the design of new transformations to the improvement to known reactions--are highlighted in this Review. The application of platinum- and gold-catalyzed transformations in natural product synthesis is also discussed. PMID- 17427894 TI - Changes in fetal motility as a result of congenital disorders: an overview. AB - After 35 years of real-time two-dimensional sonography, can we now identify changes in fetal motility indicative of malfunction of the central nervous system in high-risk pregnancies? A literature search on motor assessment procedures (movements specified for body part (SMP), quantity, quality and behavioral state), and motor milestones obtained per fetus (with various congenital disorders) yielded 48 articles describing motility of 104 fetuses. In 67%, SMPs (especially isolated arm or leg movements, breathing and general movements) were assessed. Quantitative aspects were examined in 76%, qualitative aspects in 62% and behavioral states in two fetuses. Abnormal motility can be divided into two main subcategories: hypo- and hyperkinetic, demonstrating decreased variation in qualitative performance (reduced or increased, respectively, amplitude, speed and number of participating body parts, abnormal quantity (reduced or increased, respectively) and reduced differentiation into SMPs. Posture was affected in 40/60 hypokinetic and 4/44 hyperkinetic moving fetuses. The majority of the disorders resulted in an adverse outcome. Fourteen percent survived with a handicap, depending on the underlying disorder. The 16 disorders with hypokinetic motility had mainly an autosomal recessive etiology with no possibility of invasive prenatal diagnosis or conclusive sonographic structural anomalies, in contrast to the 17 disorders with hyperkinetic motility. Within the limitations of the studies, a deeper understanding of affected milestones in motor development can be obtained. Broadening motor assessment procedures from quantitative only to include qualitative aspects, differentiation of SMPs and behavioral states and emphasizing onset and continuity of motility before and after birth will enhance the reliability and predictive value of motility as a parameter in the assessment of fetal condition. PMID- 17427895 TI - Transvaginal ultrasound measurement of cervical length and efficacy of misoprostol in first-trimester pregnancy failure. AB - OBJECTIVE: The aim of this study was to assess the role of cervical length measurement in predicting successful treatment, by misoprostol administration, of early (first-trimester) pregnancy failure. METHOD: A prospective study was conducted of all patients who agreed to medical treatment of pregnancy failure. Cervical length and other sonographic variables were measured using pelvic ultrasound before medical treatment began. Measurements were compared between the group with successful medical treatment and the group in whom treatment failed. RESULTS: In 125 women included in the study, the success rate of misoprostol treatment was 64.8%. There were no significant differences between the groups with successful and failed treatment for cervical length (29.9 +/- 9.3 vs. 30.4 +/- 6.8 mm, P = 0.75), distance between gestational sac and 'virtual' cervical internal os (23.9 +/- 13 vs. 26.6 +/- 13 mm, P = 0.26), crown-rump length (8.7 +/ 9.7 vs. 6.7 +/- 8.6 mm, P = 0.25), or gestational sac diameter (31.3 +/- 14 vs. 30.1 +/- 15 mm, P = 0.73). CONCLUSION: Cervical length does not predict the success of misoprostol treatment of first-trimester pregnancy failure. PMID- 17427896 TI - On spirometric pulmonary function in 3- to 5-year-old children. PMID- 17427897 TI - High-frequency oscillation (HFO) prevents activation of NF-kappaB found with conventional mechanical ventilation (CMV) in surfactant-depleted rabbit lung. AB - High-frequency oscillation (HFO) has been recognized as an effective ventilatory strategy to minimize lung injury during respiratory support. Conventional mechanical ventilation (CMV) compared with HFO was shown to result in an increased number of PMNs and inflammatory cytokines in the lung lavage fluid. However how mechanical forces can be sensed by cells and converted into biochemical signals for intracellular signal transduction is still unknown. In this current study, we sought to determine whether the activation of Nuclear factor-kappa B (NF-kappaB) might be involved in the lung injury caused by CMV. Surfactant-depleted Japanese white rabbits received 1- or 4-hr CMV or 1- or 4-hr HFO. Then, activation of NF-kappaB in the lungs was assessed by conducting electrophoretic mobility shift assays (EMSA). In the experiment with whole lungs, NF-kappaB activity was much higher in the 4-hr CMV lungs than in the 4-hr HFO lungs. To clarify the origin of the cells in which NF-kappaB was activated, we did a second lung lavage at the end of ventilation and washed out the cells that had infiltrated the alveoli. The levels of NF-kappaB activity were the similar in the lungs of 4-hr HFO rabbits and in those of 4-hr CMV ones. On the other hand, NF-kappaB activity was much higher in the 4-hr CMV lungs than in the 4-hr HFO lungs in the experiment with the lung lavage fluid cells. These results show that the increase in NF-kappaB activity in the lungs of 4-hr CMV rabbits was due mainly to the cells that had infiltrated the alveoli. PMID- 17427898 TI - Cutoff value of lipid-laden alveolar macrophages for diagnosing aspiration in infants and children. AB - Chronic passage of gastric and/or alimentary material into the airways is a frequent and difficult-to-diagnose condition. Because alveolar macrophages phagocytose aspirated material, it has been suggested that their identification is a useful diagnostic method. To know the usefulness of the lipid-laden alveolar macrophages (LLAM) index as a diagnostic tool for aspiration, children from 1 month to 16 years of age were included in three groups: G-I, children with pulmonary pathology and suspicion of aspiration by clinic or image evaluation; G II, with pulmonary pathology without suspicion of aspiration; and G-III, without respiratory symptoms nor suspicion of aspiration. Bronchoalveolar lavage was obtained through bronchoscopy in G-I and G-II, and through endotracheal tube in G III, and the LLAM index (0-400) was determined. A total of 112 patients (41, 30, and 41 in G-I to III, respectively) were studied. LLAM index (mean +/- SEM) was highest in G-I (233.2 +/- 5.5), as compared with G-II (187.8 +/- 11.6, P < 0.05), and G-III (108.5 +/- 13.5, P < 0.001). However, notable overlap of LLAM values was observed between G-I and G-II, and between G-II and G-III. When patients from G-I and G-III were jointly analyzed, the area under the ROC curve for diagnosing aspiration was 0.92, with a best cutoff value of >165 (98.6% sensitivity, 78.0% specificity, 87.8% overall accuracy). LLAM index, with a cutoff value of >165 is a useful diagnostic test for aspiration when there is suspicion of this condition. However, due to its low specificity, it does not discriminate other causes of chronic lung disease. PMID- 17427899 TI - Human metapneumovirus bronchiolitis in infancy is an important risk factor for asthma at age 5. AB - BACKGROUND: Human metapneumovirus (hMPV) infection plays an important role in the pediatric respiratory infections. However, little is known about the relationship between hMPV-bronchiolitis and the development of subsequent wheezing. The aim of this study was to evaluate the outcome at third to fifth year after admission for hMPV-bronchiolitis and to compare it with children admitted for respiratory syncytial virus (RSV)-bronchiolitis and with children without lower respiratory disease in the first 2 years of life. METHODS: We studied 55 children (23 hMPV and 32 RSV acute bronchiolitis) aged 3-5 years and hospitalized during the seasons 2000-2005. Thirty-eight children admitted due to acute rotavirus gastroenteritis in the same seasons, and without bronchiolitis during infancy, were also analyzed. Clinical data were collected through structured clinical interviews at the follow-up visit. Skin prick tests to food and inhaled allergens were performed. RESULTS: Asthma was significantly more frequent in children with hMPV-bronchiolitis (odds ratio (OR) = 5.21), RSV-bronchiolitis (OR = 4.68), allergic rhinitis (OR = 1.7), and siblings with atopy (OR = 1.75). In the multivariate analyses, hMPV was the most important risk factor for asthma in preschool years (OR = 15.9), followed by RSV-bronchiolitis (OR = 10.1) and allergic rhinitis (OR = 4.9). No differences were found with respect to allergic rhinitis, atopic dermatitis, food allergy, proportion of positive prick tests, family history of atopy, and asthma. CONCLUSIONS: hMPV-bronchiolitis in infancy was strongly associated with asthma as well as other forms of bronchial obstructive disease at third and fifth year of life. This association is, at least, as strong as the association observed with RSV infections. PMID- 17427900 TI - Design of molecular logic devices based on a programmable DNA-regulated semisynthetic enzyme. PMID- 17427901 TI - H2O2 oxidations catalyzed by an iron(III) corrolazine: avoiding high-valent iron oxido species? PMID- 17427902 TI - Synthesis of 2,3-disubstituted indoles by a rhodium-catalyzed aromatic amino Claisen rearrangement of N-propargyl anilines. PMID- 17427903 TI - Stents (grafting) should be the primary treatment for symptomatic superficial femoral artery disease. PMID- 17427904 TI - Magnetic navigation: a pivotal technology. PMID- 17427905 TI - Emotional numbing weakens abused inner-city women's resiliency resources. AB - Conservation of resources theory (S. E. Hobfoll, 1988, 1999) hypothesizes that loss of resiliency resources can contribute to abused women's posttraumatic stress disorder (PTSD) symptoms, which, in turn, contribute to a further loss of resources, which can make abused women even more vulnerable to future stressors. This study investigates the impact of PTSD symptoms on abused women's future loss of resources-resources that women both value and need to aid their ongoing adjustment. Posttraumatic stress disorder symptoms contributed to future resource loss in abused women, even when controlling for the effects of prior resource loss and depression. Emotional numbing symptoms of PTSD accounted most for women's resource loss. Findings highlight the importance of research and intervention that more directly examines the link between emotions and resource loss. PMID- 17427906 TI - Mediators of control beliefs, stressful life events, and adaptive behavior in school age children: the role of appraisal and social support. AB - The authors examine the role of appraisal and social support as mediators of the relation between control beliefs and adaptive behavioral outcome. Using the responses from 297 children, ages 8 to 12 years old, the results suggest two significant mediational pathways. Social support was a mediator of the relation between unknown control for negative events and adaptive behavior and the relation between unknown control for positive events and adaptive behavior. Negative appraisal demonstrated no mediation relations. The role of social support and negative appraisal in the display of adaptive behavior and the implications for further model testing are discussed. PMID- 17427907 TI - Quality of parental relationships among persons with a lifetime history of posttraumatic stress disorder. AB - Several studies of combat veterans have examined the relationship between parental satisfaction and PTSD symptoms. These studies found that numbing is associated with substantial decrements in parent-child relationship quality. The current study extends previous work by assessing the effect of PTSD on parent child relationships in a nationally representative sample of civilian men and women with PTSD resulting from a broad range of trauma. It was hypothesized that PTSD avoidance/numbing symptoms would be predictive of parent-child relationship quality and parent-child conflict. Moreover, these relationships are predicted to hold after controlling for a broad range of support-related variables and work/finance related variables. As hypothesized, after controlling for number of children and respondent-initiated domestic violence, numbing was predictive of increased parent-child aggression. PMID- 17427908 TI - Posttraumatic appraisals in the development and persistence of posttraumatic stress symptoms. AB - Cognitive models of posttraumatic stress disorder (PTSD) posit that appraisal plays an important role in the development and persistence of PTSD. This study examined posttraumatic appraisals and their relationship to the development and course of PTSD symptoms. Two hundred fifty-three injury survivors were assessed for PTSD symptoms and posttraumatic cognitions across a 12-month period. A path analytic modeling approach showed that posttraumatic appraisals were important direct and indirect predictors of later PTSD severity. The findings suggest that appraisals made in the aftermath of trauma have a significant influence on subsequent psychological adjustment. PMID- 17427909 TI - Intrusive memories and ruminations related to violent crime among young offenders: phenomenological characteristics. AB - Very little is known about the nature of perpetrator's memories of violent crime. The authors conducted semistructured interviews with a representative sample of 105 young offenders convicted of serious violence, assessing intrusive memories, ruminations, and symptoms of posttraumatic stress disorder related to their violent crime. Forty-eight (46%) participants described significant intrusive memories of the assault, and 38 (36%) reported ruminations related to the assault. Ethnic origin and historical variables explained 19% of the variance of posttraumatic stress disorder symptom severity; intrusion and rumination characteristics added an additional 48% explained variance. The intrusive memories tended to concern the moment when the event turned for the worse for the perpetrator. The findings have implications for risk assessment and therapeutic interventions for violent offenders. PMID- 17427910 TI - Confirmatory factor analysis of posttraumatic stress symptoms in Cambodian refugees. AB - This study used confirmatory factor analysis to examine the factor structure of the Cambodian version of the Harvard Trauma Questionnaire in 488 Cambodian refugees residing in the United States. Five alternative conceptual models were compared. A model with four correlated factors reflecting symptoms of re experiencing, avoidance, emotional numbing, and hyperarousal provided the best fit to these data. The avoidance and emotional numbing clusters demonstrated distinct associations with depression symptoms, providing some evidence of the construct validity of the four-factor solution. PMID- 17427912 TI - Aggression among combat veterans: relationships with combat exposure and symptoms of posttraumatic stress disorder, dysphoria, and anxiety. AB - Prior research has revealed heightened aggressive behavior among veterans with PTSD. This study tested a model examining the interrelationships among combat exposure, posttraumatic stress disorder (PTSD) symptoms, dysphoric symptoms, and anxiety symptoms in predicting aggressive behavior in a sample of 265 male combat veterans seeking diagnostic assessment of PTSD. Combat exposure was indirectly associated with aggression primarily through its relationship with PTSD symptoms. Symptoms of PTSD were directly related to aggression, and indirectly related to aggression through dysphoric symptoms. Results highlight the role of PTSD symptoms and dysphoric symptoms with respect to aggressive behavior among this population, and suggest the relevance of aggression theory to the study of combat veterans. PMID- 17427913 TI - The Criterion A problem revisited: controversies and challenges in defining and measuring psychological trauma. AB - The Criterion A problem in the field of traumatic stress refers to the stressor criterion for posttraumatic stress disorder (PTSD) and involves a number of fundamental issues regarding the definition and measurement of psychological trauma. These issues first emerged with the introduction of PTSD as a diagnostic category in the Diagnostic and Statistical Manual of Mental Disorders, Third Edition (DSM-III; American Psychiatric Association, 1980) and continue to generate considerable controversy. In this article, the authors provide an update on the Criterion A problem, with particular emphasis on the evolution of the DSM definition of the stressor criterion and the ongoing debate regarding broad versus narrow conceptualizations of traumatic events. PMID- 17427914 TI - Randomized clinical trial for treatment of chronic nightmares in trauma-exposed adults. AB - Nightmares and sleep disturbance are fundamental concerns for victims of trauma. This study examined the efficacy of a manualized cognitive-behavioral treatment (CBT) for chronic nightmares in trauma-exposed individuals via a randomized clinical trial. Participants were randomly assigned to a treatment group or wait list control group, with 27 participants completing the treatment. At the 6-month follow-up assessment, 84% of treated participants reported an absence of nightmares in the previous week. Significant decreases were also reported in symptoms of depression and posttraumatic stress, fear of sleep, and number of sleep problems, while sleep quality and quantity improved. The present study adds to the growing literature indicating this brief CBT as a first-line treatment for trauma-exposed individuals with chronic nightmares. PMID- 17427915 TI - The human bovine arch--a common misnomer. PMID- 17427916 TI - Visualization of iliac and proximal femoral artery hemodynamics using time resolved 3D phase contrast MRI at 3T. AB - PURPOSE: To demonstrate the feasibility of time-resolved 3D MR velocity mapping at 3 Tesla for the visualization of vascular hemodynamics in normal iliac and femoral arteries. MATERIALS AND METHODS: Electrocardiographically (ECG) synchronized three-dimensional (3D) CINE phase-contrast MRI with three directional flow encoding was adapted to analyze flow in peripheral arteries at 3T. Visualization of peripheral arterial hemodynamics within the acquired data volume included 3D streamlines and time-resolved 3D particle traces within the major vessels and localized analysis of flow profiles using 2D-vector graphs. Data was visually compared to results from color-coded duplex ultrasound (US). RESULTS: Global and detailed local blood flow characteristics were successfully analyzed in all subjects. In agreement with US findings, normal laminar flow patterns without flow acceleration or disturbances were visualized in all healthy individuals. In an exemplary patient measurement multiple segmental flow accelerations could be demonstrated. MRI additionally revealed complex helical flow alterations distal to a moderate stenosis. CONCLUSION: Due to the full spatial and temporal coverage of the arteries of interest, 3D CINE phase contrast MRI at 3T is a promising tool for the evaluation of vascular hemodynamics in peripheral arteries. Future methodological improvements will be directed to improve spatial and temporal resolution as well as quantitative data analysis. Moreover, the technique will have to be evaluated in patients in comparison to standard diagnostic tools. PMID- 17427918 TI - Characterization of a novel SPG3A deletion in a French-Canadian family. AB - Hereditary spastic paraplegias (HSPs) are characterized by progressive lower limb spasticity and weakness. Mutations in the SPG3A gene, which encodes the large guanosine triphosphatase atlastin, are the second most common cause of autosomal dominant hereditary spastic paraplegia. In a large SPG3A screen of 70 hereditary spastic paraplegia subjects, a novel in-frame deletion, p.del436N, was identified. Characterization of this deletion showed that it affects neither the guanosine triphosphatase activity of atlastin nor interactions between atlastin and spastin. Interestingly, immunoblot analysis of lymphoblasts from affected patients demonstrated a significant reduction in atlastin protein levels, supporting a loss-of-function disease mechanism. PMID- 17427919 TI - Estradiol attenuates hyperoxia-induced cell death in the developing white matter. AB - OBJECTIVE: Periventricular leukomalacia is the predominant type of brain injury in preterm infants underlying the development of cerebral palsy. Periventricular leukomalacia has its peak incidence at 23 to 32 weeks postconceptional age characterized by extensive oligodendrocyte migration and maturation. Oxygen toxicity has been identified as a possible contributing factor to the pathogenesis of cerebral palsy in survivors of preterm birth. 17beta-estradiol (E2) is important for the development and function of the central nervous system. Furthermore, neuroprotective properties have been attributed to estrogens. We examined the effect of E2 on hyperoxia-induced cell death in the developing white matter in the rat brain. METHODS: Six-day-old (P6) rat pups, the immature oligodendroglial cell line (OLN-93), and primary oligodendrocyte cultures were subjected to 80% O(2) in the presence or absence of E2 (600 microg/kg intraperitoneally in vivo, 10(-6)-10(-10)M in vitro). Cell counts and lactate dehydrogenase assay were used to assess cell survival. Immunoblot analysis was used for detection of estrogen receptor expression and investigation of apoptotic signaling pathways. White matter injury was assessed by myelin basic protein immunocytochemistry at P11. RESULTS: E2 produced significant dose-dependent protection against oxygen-induced apoptotic cell death in primary oligodendrocytes. Treatment with E2 prevented hyperoxia-induced proapoptotic Fas upregulation and caspase-3 activation. Finally, E2 antagonized hyperoxia-induced inactivation of extracellular signal-regulated kinase 1 and 2 and Akt, key kinases of the mitogen-activated protein kinase and phosphatidylinositol 3-kinase cell survival promoting pathways, respectively. Loss of myelin basic protein labeling was seen in P11 pups after oxygen exposure, and E2 attenuated this injury. INTERPRETATION: These results suggest a possible role for estrogens in the prevention of neonatal oxygen-induced white matter injury. PMID- 17427920 TI - Imaging correlates of axonal swelling in chronic multiple sclerosis brains. AB - OBJECTIVE: T2-weighted magnetic resonance imaging is a sensitive tool for monitoring progression of multiple sclerosis, but it does not provide information on the severity of the underlying tissue damage. Measurement of T1 hypointensities and magnetization transfer ratio (MTR) can potentially distinguish lesions with more severe tissue damage. The objective of this study was to use image-guided pathology to determine histological differences between lesions that are abnormal only on T2-weighted images versus lesions that are abnormal on T2-weighted, T1-weighted, and MTR images. METHODS: A total of 110 regions were selected from postmortem magnetic resonance images of 10 multiple sclerosis patients. Regions were classified into three magnetic resonance imaging defined categories: normal-appearing white matter; abnormal on T2-weighted image only (T2-only); and abnormal on T2-weighted, T1-weighted, and MTR images (T2T1MTR). Myelin status, lesion activity, astrocytosis, serum protein distribution, axonal area, and axonal loss were evaluated histopathologically. RESULTS: Comparisons between groups showed that T2T1MTR regions were more likely to be demyelinated (83% compared with 55% of T2-only regions) and more likely to be chronic inactive lesions (68% compared with 0% of demyelinated T2-only regions). There was no difference between T2-only and T2T1MTR regions in axonal area, but there was a significant difference in axonal count, indicating that axons in the T2T1MTR regions were enlarged relative to those in T2-only regions. INTERPRETATION: Axonal swelling and axonal loss were major pathological features that distinguish T2T1MTR regions from T2-only regions. PMID- 17427922 TI - Niger-Congo speaking populations and the formation of the Brazilian gene pool: mtDNA and Y-chromosome data. AB - We analyzed sequence variation in the mitochondrial DNA (mtDNA) hypervariable segment I (HVS-I) from 201 Black individuals from two Brazilian cities (Rio de Janeiro and Porto Alegre), and compared these data with published information from 21 African populations. A subset of 187 males of the sample was also characterized for 30 Y-chromosome biallelic polymorphisms, and the data were compared with those from 48 African populations. The mtDNA data indicated that respectively 69% and 82% of the matrilineages found in Rio de Janeiro and Porto Alegre originated from West-Central/Southeast Africa. These estimates are in close agreement with historical records which indicated that most of the Brazilian slaves who arrived in Rio de Janeiro were from West-Central Africa. In contrast to mtDNA, Y-chromosome haplogroup analysis did not allow discrimination between places of origin in West or West-Central Africa. Thus, when comparing these two major African regions, there seems to be higher genetic structure with mtDNA than with Y-chromosome data. PMID- 17427923 TI - Infant carrying: the role of increased locomotory costs in early tool development. AB - Among the costs of reproduction, carrying one's infant incurs one of the greatest drains on maternal energy, simply because of the added mass alone. Because of the dearth of archaeological evidence, however, how early bipeds dealt with the additional cost of having to carry infants who were less able to support their body weight against gravity is not particularly well understood. This article presents evidence on the caloric drain of carrying an infant in one's arms versus having a tool with which to sling the infant and carry her passively. The burden of carrying an infant in one's arms is on average 16% greater than having a tool to support the baby's mass and seems to have the potential to be a greater energetic burden even than lactation. In addition, carrying a baby in one's arms shortens and quickens the stride. An anthropometric trait that seems to offset some of the increased cost of carrying a baby in the arms is a wider bi trochanteric width. PMID- 17427924 TI - The slow and unnoticed changes in the funding mix. PMID- 17427925 TI - mtDNA from hair and nail clarifies the genetic relationship of the 15th century Qilakitsoq Inuit mummies. AB - The 15th century Inuit mummies excavated at Qilakitsoq in Greenland in 1978 were exceptionally well preserved and represent the largest find of naturally mummified specimens from the Arctic. The estimated ages of the individuals, their distribution between two adjacent graves, the results of tissue typing, and incomplete STR results led researchers to conclude that the eight mummies formed two distinct family groups: A grandmother (I/5), two daughters (I/3, I/4), and their two children (I/1, I/2) in one grave, and two sisters (II/6, II/8) and a daughter (II/7) of one of them in the other. Using mtDNA from hair and nail, we have reanalyzed the mummies. The results allowed the unambiguous assignment of each of the mummies to one of three mtDNA haplogroups: A2b (I/5); A2a (I/2, I/3, II/6, II/8); A2a-311 (I/1, I/4, II/7), excluded some of the previous relations, and pointed to new ones. I/5 is not the grandmother/mother of the individuals in Grave I, and she is not maternally related to any of the seven other mummies; I/3 and I/4 are not sisters and II/7 is neither the daughter of II/6 nor of II/8. However, I/1 may be the child of either I/4 or II/7 and these two may be sisters. I/2 may be the son of I/3, who may be the daughter of either II/6 or II/8, and these two may be sisters. The observation of haplogroups A2a and A2b amongst the 550-year-old Inuit puts a lower limit on the age of the two lineages in Greenland. PMID- 17427926 TI - A bioarcheological study of maxillary sinusitis. AB - Maxillary sinusitis was studied as an indicator of poor air quality. Seven skeletal samples were examined from North America, England, and Nubia, and selected to represent different geographic locations, environments, and subsistence economies. Frequency rates varied from 17.2 to 51.5% of individuals affected with one or both sinuses preserved. Hardin Village had the highest frequency (51.5%), followed by the Aleuts (42.9%), "Illinois" (38.6%), Indian Knoll (38.5%), Kulubnarti (21.8%), Christchurch, Spitalfields (18.0%), and "South Dakota" (17.2%). Male frequencies ranged from 16.7 to 36.7%, but the female frequency ranged more widely from 18.0 to 76.5%. At most sites female rates exceeded male. The effect of urban and rural environment on sinusitis occurrence, and also subsistence economy, biological sex, and social status were explored, and comparative sites also considered; urban agricultural sites had a mean frequency of 48.5%, rural agricultural sites had a mean frequency of 45.0%, and hunter-gatherer sites had a mean frequency of 40.0%. In the urban sites male and female frequencies were near equal, but in the rural agricultural and hunter gatherer sites female frequencies exceeded male frequencies. Dental disease was not found to have much impact on sinusitis frequency. The importance of the link between poor air quality and respiratory health is highlighted in clinical studies in both developed and developing countries, but also in bioarcheological studies. PMID- 17427927 TI - North Indian Muslims: enclaves of foreign DNA or Hindu converts? AB - The mtDNA composition of two Muslim sects from the northern Indian province of Uttar Pradesh, the Sunni and Shia, have been delineated using sequence information from hypervariable regions 1 and 2 (HVI and HVII, respectively) as well as coding region polymorphisms. A comparison of this data to that from Middle Eastern, Central Asian, North East African, and other Indian groups reveals that, at the mtDNA haplogroup level, both of these Indo-Sunni and Indo Shia populations are more similar to each other and other Indian groups than to those from the other regions. In addition, these two Muslim sects exhibit a conspicuous absence of West Asian mtDNA haplogroups suggesting that their maternal lineages are of Indian origin. Furthermore, it is noteworthy that the maternal lineage data indicates differences between the Sunni and Shia collections of Uttar Pradesh with respect to the relative distributions of Indian specific M sub-haplogroups (Indo Shia > Indo Sunni) and the R haplogroup (Indo Sunni > Indo Shia), a disparity that does not appear to be related to social status or geographic regions within India. Finally, the mtDNA data integrated with the Y-chromosome results from an earlier study, which indicated a major Indian genetic (Y-chromosomal) contribution as well, suggests a scenario of Hindu to Islamic conversion in these two populations. However, given the substantial level of the African/Middle Eastern YAP lineage in the Indo-Shia versus its absence in the Indo-Sunni, it is likely that this conversion was somewhat gender biased in favor of females in the Indo-Shia. PMID- 17427928 TI - Orientation of the human sacrum: anthropological perspectives and methodological approaches. AB - Discovering the nature of sacral orientation is of considerable anthropological importance. Therefore, this study aims at presenting a new anthropologically based definition for sacral anatomical orientation (SAO) angle, establishing standards of SAO for human population; examining the relationship between pelvic incidence (PI) and SAO; and associating SAO with demographic parameters. The study population consisted of 424 adult and 14 sub-adult (13-18 years, for SAO only) pelvises. Sacral orientation was measured using two different definitions: a) SAO is the angle created between the intersection of a line running parallel to the superior surface of the sacrum and a line running between the anterior superior iliac spine (ASIS) and the anterior-superior edge of the symphysis pubis; b) PI is the angle created between the perpendicular to the sacral plate at its midpoint and the line connecting this point to the middle of the axis of the acetabulum. SAO was measured using a specially designed mechanical measurement tool and a 3D digitizer. PI was measured via the 3D digitizer. The methods developed by us for measuring SAO and PI in skeletal material are valid and reliable. SAO and PI measures were highly correlated (r = -0.824, P < 0.001). The average SAO was 49.01 degrees (SD = 10.16), and the average PI 54.08 degrees (SD = 12.64). SAO was independent of ethnicity and sex, yet age dependent. This study establishes a methodology for estimating SAO and PI in skeletal material and furnishes the anthropological milieu with base line data regarding these parameters. Future studies in human evolution can greatly benefit from this study. PMID- 17427929 TI - Understanding the adaptive value of diagonal-sequence gaits in primates: a comment on Shapiro and Raichlen, 2005. PMID- 17427930 TI - Free radical generation during the activation of hemolymph prepared from the homopteran Dactylopius coccus. AB - Superoxide anion (O(-) (2)) and nitric oxide (NO) generation in Dactylopius coccus hemolymph obtained by perfusion and activated with zymosan was studied. Activated hemolymph reduced 3-[4,5 dimethylthiazolil-2]-2,5-diphenyl tetrazolium bromide. This reduction was prevented by superoxide dismutase (SOD) indicating O( ) (2) generation. This activity was dependent on temperature, and hemolymph incubated at 75 degrees C lost its activity. Chromatocytes incubated with zymosan released their content and produced O(-) (2). Activated hemolymph also produced NO and this activity was prevented in the presence of NG-nitro-L-arginine methyl ester, suggesting that nitric oxide synthase (NOS) might be present in D. coccus hemolymph. The probable source of O(-) (2) in the D. coccus hemolymph is the anthraquinone oxidation, since commercial carminic dye produced O(-) (2) during its oxidation by Agaricus bisporus tyrosinase. Gram+ Micrococcus luteus exposed to activated hemolymph were killed in vitro, and addition of NG-nitro-L-arginine methyl ester and D-Mannitol (a hydroxyl radical scavenger) prevented their killing. The cytotoxic effect produced by the activated hemolymph was not observed with the Gram- bacteria Serratia marcescens. These results suggest that D. coccus activated hemolymph generates reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) that may limit M. luteus growth. PMID- 17427931 TI - New isoforms of odorant-binding proteins and potential semiochemicals of locusts. AB - To obtain more information on the elements of chemical communication in the migratory locust (Locusta migratoria) (Orthoptera: Acrididae), we have searched for additional odorant-binding proteins (OBPs) and for volatiles in the feces that could represent potential semiochemicals for this species. A two-dimensional electrophoretic (2DE) analysis of an antennal extract showed only three closely positioned spots that were recognized by the antiserum against locust OBP. Three genes were also identified using PCR and 5'RACE-PCR approaches, encoding isoforms differing from each other for a single amino acid substitution. The gas chromatographic-electroantennogram (GC-EAD) headspace analysis of a feces sample revealed the presence of several compounds that elicited dose-dependent electrophysiological responses in the antennae of both sexes. Most of these compounds are different from those identified in the feces of the desert locust (Schistocerca gregaria) and reported to be behaviorally active. Ligand-binding experiments performed with such volatiles and recombinant OBP did not show affinity, thus indicating that the binding pocket of OBP requires larger molecules than those so far identified. PMID- 17427932 TI - Identification of Y chromosomal PCR marker and production of a selected strain for molecular sexing in the brown planthopper, Nilaparvata lugens. AB - A laboratory colony was established in order to enable molecular sexing in premature stages in the brown planthopper, Nilaparvata lugens. We found four male specific amplified fragment length polymorphisms (AFLPs) in the planthopper, and sequenced one of the AFLPs along with its 5' flanking region (1,423 bp in total). PCR primers were designed based on the nucleotide sequence information so that the PCR product was present in male planthoppers and absent in female planthoppers. However, we could not completely distinguish males from females, because the PCR amplification product was absent in some of the males screened. We, therefore, established a laboratory colony, in which all males carried this sequence. We can directly sex pre-adult stages in this colony using our PCR primers, making this strain of considerable value for studies that require sex separation in egg and nymphal stages. PMID- 17427933 TI - Bacteria and PAMPs activate nuclear factor kappaB and Gro production in a subset of olfactory ensheathing cells and astrocytes but not in Schwann cells. AB - The primary olfactory nerves provide uninterrupted conduits for neurotropic pathogens to access the brain from the nasal cavity, yet infection via this route is uncommon. It is conceivable that olfactory ensheathing cells (OECs), which envelope the olfactory nerves along their entire length, provide a degree of immunological protection against such infections. We hypothesized that cultured OECs would be able to mount a biologically significant response to bacteria and pathogen-associated molecular patterns (PAMPs). The response of OECs to Escherichia coli (E. coli) and various PAMPs was compared to that of Schwann cells (SCs), astrocytes (ACs), and microglia (MG). A subset of OECs displayed nuclear localization of nuclear factor kappaB), an inflammatory transcription factor, after treatment with E. coli (20% +/- 5%), lipopolysacchride (33% +/- 9%), and Poly I:C (25% +/- 5%), but not with peptidoglycan or CpG oligonucleotides. ACs displayed a similar level of activation to these treatments, and in addition responded to peptidoglycan. The activation of OECs and ACs was enhanced by coculture with MG (56% +/- 16% and 85% +/- 13%, respectively). In contrast, SCs did not respond to any treatment or to costimulation by MG. Immunostaining for the chemokine Gro demonstrated a functional response that was consistent with NF kappaB activation. OECs expressed mRNA for Toll-like receptors (TLRs) 2 and 4, but only TLR4 protein was detected by Western blotting and immunohistochemistry. The results demonstrate that OECs possess the cellular machinery that permits them to respond to certain bacterial ligands, and may have an innate immune function in protecting the CNS against infection. PMID- 17427934 TI - Regulation of pheromone biosynthesis in the "Z strain" of the European corn borer, Ostrinia nubilalis. AB - The regulation of pheromone biosynthesis by the neuropeptide PBAN in the Z strain of the European corn borer, Ostrinia nubilalis, was investigated using labeled intermediates. Injection of radiolabeled acetate showed PBAN did not influence the de novo synthesis of saturated fatty acids in the gland. When deuterium labeled myristic acid was topically applied to the gland, females injected with PBAN produced more labeled pheromone than did control females, indicating that PBAN controls one of the later steps of pheromone biosynthesis. Although more myristic acid was Delta11-desaturated in the gland in the presence of PBAN, this was counterbalanced by less Delta11-desaturation of palmitic acid, indicating that desaturase activity did not change overall. This change in flux of myristic acid through to pheromone was shown to be caused by increased reduction of fatty acid pheromone precursors occurring in the presence of PBAN. PMID- 17427935 TI - Effects of gamma-irradiation on midgut proteolytic activity of the mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae). AB - The Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann), is a key pest of citrus in Spain because of significant yield losses and to quarantine restrictions. Biologically based control methods, such as the Sterile Insect Technique (SIT), which relies on the sterilization by irradiation of large numbers of insects, is gaining an increasing role in the control of medfly in Mediterranean areas. However, gamma-irradiation might damage the midgut epithelium cells, causing a lowering of nutritive assimilation that can negatively affect adult performance. Irradiation effects on digestive physiology are well established for a number of insect pests, but there is no information on medfly. Our aim was to determine the effects of gamma-irradiation on C. capitata digestive protease activity. Both larvae and adults were found to use a similar proteolytic system based on aspartyl-, trypsin-, chymotrypsin-, amino peptidase-, and carboxypeptidase A- and B-like activities. Pupae of the Vienna-7 (tsl) strain were irradiated at 70 or 140 Gy, two days before emergence, and the adults fed during 5 days on sugar-protein (4:1) diets. Protease activity was measured in midgut extracts and compared with males non-irradiated reared in the same conditions. The results showed that the irradiation doses tested had no effect on the digestive proteolytic activities of medfly adults. Moreover, the longevity of irradiated medflies at the highest dose (140 Gy) was similar to that of controls. PMID- 17427936 TI - High dose levodopa therapy is not toxic in multiple system atrophy: experimental evidence. AB - Levodopa is generally regarded the first choice therapy for parkinsonism associated with multiple system atrophy (MSA-P). However, MSA-P patients often show a poor or unsustained levodopa response which inflicts high dose therapy. This is generally attributed to progressive striatal degeneration with loss of dopamine receptors. Experimental evidence suggests that dopaminergic stimulation may accelerate the striatal disease process in MSA, possibly by pro-oxidative mechanisms. Intact nigrostriatal dopamine release augments striatal lesion size in the unilateral nigral and striatal double lesion rat model of MSA-P. Further, neuronal vulnerability to exogenous oxidative stress is increased in a transgenic MSA mouse model with oligodendroglial alpha-synuclein inclusions. The aim of the present study was to analyze whether high dose levodopa delivery in the transgenic MSA model is associated with neurotoxicity exacerbated by the presence of oligodendroglial alpha-synuclein inclusion pathology. Control and transgenic MSA mice underwent pulsatile treatment with either vehicle, low or high dose levodopa for a period of 1 month. Behavioral and neuropathological indices failed to show evidence for neurotoxic effects of high-dose levodopa in this alpha synuclein transgenic MSA model. These findings support the idea that high dose levodopa therapy in MSA is not detrimental to the underlying neuropathological process. PMID- 17427937 TI - Successful treatment of truncal myoclonus. PMID- 17427938 TI - Distinct distribution of autosomal dominant spinocerebellar ataxia in the Mexican population. AB - Dominant ataxias show wide geographic variation. We analyzed 108 dominant families and 123 sporadic ataxia patients from Mexico for mutations causing SCA1 3, 6-8, 10, 12, 17 and DRPLA. Only 18.5% of dominant families remained undiagnosed; SCA2 accounted for half (45.4%), followed by SCA10 (13.9%), SCA3 (12%), SCA7 (7.4%), and SCA17 (2.8%). None had SCA1, 6, 8, 12 or DRPLA. Among sporadic cases, 6 had SCA2 (4.9%), and 2 had SCA17 (1.6%). In the SCA2 patients we identified 6 individuals with the rare (CAG)(33) allele, 2 of whom showed early onset ataxia. The distribution of dominant ataxia mutations in Mexicans is distinct from other populations. PMID- 17427939 TI - Management of status dystonicus: our experience and review of the literature. AB - Status dystonicus (SD) is a life threatening disorder that develops in patients with both primary and secondary dystonia, characterized by acute worsening of symptoms with generalized and severe muscle contractions. To date, no information is available on the best way to treat this disorder. We review the previously described cases of SD and two new cases are reported, one of which occurring in a child with static encephalopathy, and the other one in a patient with pantothenate kinase-associated neurodegeneration. Both patients were admitted to an intensive care unit and treated with midazolam and propofol. This approach proved to be useful in the former while the progressive nature of the dystonia of the second patient required the combination of intrathecal baclofen infusion and bilateral pallidal deep brain stimulation. We believe that a rapid and aggressive approach is justified to avoid the great morbidity and mortality which characterize SD. Our experience, combined with the data available in the literature, might permit to establish the best strategies in managing this rare and severe condition. PMID- 17427940 TI - Levodopa-induced dyskinesias. AB - Levodopa-induced dyskinesias (LID) are common and difficult to treat. This review focuses on three issues related to LID: clinical features, classification and rating, pathophysiology and pathogenesis, and management. The three primary clinical syndromes are OFF-period dystonia, peak-dose dyskinesia, and diphasic dyskinesia. Several other forms also occur, making the evaluation and choice of treatment complicated. A core component of the pathophysiology of LID is overactivity of the direct striatal output pathway. This pathway provides a direct GABAergic connection by which the striatum inhibits the output regions of the basal ganglia, i.e., the internal globus pallidus and the substantia nigra pars reticulata. Altering dopaminergic dosing and timing can abate dyskinesias, but usually impact the control of parkinsonism. Putative therapies to reduce the problem of dyskinesias could focus on the glutamatergic, GABAergic, alpha2 adrenergic, serotonergic (5HT1A, 5HT2A), opioid, histamine H3, adenosine A2A receptors, the monoamine transport or cannabinoid CB1 receptors systems. The only currently available drug with an evidence-based recommendation on efficacy for dyskinesia is amantadine. Therapy goals include the prevention of dyskinesia and treatment of dyskinesias that are troublesome clinically. New rating measures to assess severity and disability related to dyskinesia are in the process of development and clinimetric testing. PMID- 17427941 TI - Prevalence and clinical features of common LRRK2 mutations in Australians with Parkinson's disease. AB - We determined the prevalence of two common leucine-rich repeat kinase 2 (LRRK2) gene mutations in Australian patients with Parkinson's disease (PD). Of 830 affected patients, eight were heterozygous for the G2019S mutation, and two were heterozygous for the R1441H (4,322 G > A) mutation. In addition, one familial patient had a novel A1442P (4,324 G > C) mutation. Haplotype analysis showed that all LRRK2 G2019S-positive individuals carried the common founder haplotype 1 and a putative founder haplotype for the R1441H mutation carriers. Clinically, patients with LRRK2 mutations had typical levodopa responsive Parkinsonism with tremor being the commonest presenting feature. Patients with the G2019S mutation in our series had a similar age of onset of symptoms when compared with patients with other LRRK2 mutations or sporadic PD, although they were more likely to have a family history of PD (2.4% of Australian patients with familial PD and 0.3% of Australian patients with sporadic PD). Our results demonstrate that the G2019S mutation carriers share the same ancestors who migrated to Australia originally from Europe and that other LRRK2 mutations (R1441H and A1442P) can be found in this population. PMID- 17427942 TI - Myocardial sympathetic degeneration correlates with clinical phenotype of Parkinson's disease. AB - In idiopathic Parkinson's disease (PD), different clinical subtypes are distinguished due to predominant motor symptoms: a tremor-dominant type (TDT), an akinetic rigid type (ART), and a mixed type (MT). We compared myocardial sympathetic innervation, measured by MIBG scintigraphy, in different subtypes of PD at early and advanced stages of PD. We applied MIBG scintigraphy in 102 patients with PD. About 57 patients were at Hoehn and Yahr (H&Y) stage 1, 22 at H&Y stage 2, and 23 at H&Y stages 3 and 4. For quantification of myocardial MIBG uptake, the heart-to-mediastinum (H/M) count-ratio was calculated. At all H&Y stages, myocardial MIBG uptake was significantly higher in TDT patients than in ART or MT patients (P < 0.05; ANOVA). Furthermore, at each H&Y stage, myocardial MIBG uptake correlated significantly with severity of hypokinesia (P < 0.05; Spearman's correlation) and rigidity (P < 0.05), but not with severity of resting or postural tremor. The significant correlation between myocardial sympathetic degeneration and severity of hypokinesia and rigidity suggests that myocardial sympathetic degeneration and hypokinetic-rigid symptoms develop in a closely coupled manner in early as well as advanced PD. No such correlation can be found between myocardial sympathetic degeneration and parkinsonian tremor. PMID- 17427943 TI - Co-morbidity and the frequency of general practitioner consultations in Parkinson's disease in the United Kingdom. PMID- 17427944 TI - Effect of steroid hormones on Bufo arenarum oviduct. Ultrastructural study. AB - The endocrine regulation of the mucosa of the oviductal pars convoluta was analyzed by ultrastructural studies demonstrating that ovariectomy, together with a decrease in ovarian steroids circulating levels, caused a marked regression in this portion of Bufo arenarum oviduct. Twenty-five days after ovariectomy, a decrease in the depth of the epithelial and glandular layers was observed due to the notable loss of secretory cells, whose number was clearly smaller than in nonovariectomized females. The remaining secretory cells showed involution signs, with few secretory granules in their cytoplasm, little endoplasmic reticulum near poorly developed Golgi complexes and a large amount of lipid droplets. Cells in an advanced autolysis state were found in the lumen. These characteristics evidence a nonfunctional state of the pars convoluta. Treatment with 5alpha dihydrotestosterone (DHT) completely reversed the ovariectomy effect, inducing pars convoluta growths and restoring the characteristics of epithelial and glandular secretory cells in the whole pars convoluta, with micrographs similar to the control. These same effects were observed after treatment with estradiol 17beta (E2), progesterone (P) o E(2)+P in the glandular layer of the whole pars convoluta, but only in the epithelial layer of the most anterior region of this duct. In the secretory cells of other segments these treatments induced the formation of granules of high electron density and homogeneous aspect. Each steroid had a particular effect on the pars convoluta. Although E2 and DHT induced the development of the organoids involved in the proteins biosynthesis, P and DHT acted as secretagogues. PMID- 17427945 TI - Diverse effects of Stat1 on the regulation of hsp90alpha gene under heat shock. AB - Stat1 has been known as a regulator of gene expression and a mediator of IFNgamma signaling in mammalian cells, while its effect in a heat shock response remains unclear. We used RNAi knockdown, point mutations, ChIP and promoter activity assays to study the effect of Stat1 on the heat-shock induction of the hsp90alpha gene under heat shock conditions. We found that Stat1 regulates the heat shock induction of its target genes, the hsp90alpha gene in a heat shock response while the constitutive activity of the gene remains unaffected. The result of Stat1 in complex with Stat3 and HSF1 that bound at the GAS to lead a moderate heat shock induction was designated as an "intrinsic" induction of the hsp90alpha gene. Additionally a reduced or an elevated level of heat shock induction was also controlled by the Stat1 on hsp90alpha. These diverse effects on the hsp90alpha gene were a "reduced" induction with over-expressed Stat1 elicited by transfection of wild-type Stat1 or IFNgamma treatment, bound at the GAS as homodimer; and an "enhanced" heat shock induction with a mutation-mediated prohibition of Stat1/GAS binding. In conclusion, the status and efficacy of Stat1 bound at the GAS of its target gene are pivotal in determining the impact of Stat1 under heat shock. The results provided the first evidence on the tumor suppressor Stat1 that it could play diverse roles on its target genes under heat shock that also shed lights on patients with fever or under thermotherapy. PMID- 17427946 TI - Molecular definitions of fatty acid hydroxylases in Arabidopsis thaliana. AB - Towards defining the function of Arabidopsis thaliana fatty acid hydroxylases, five members of the CYP86A subfamily have been heterologously expressed in baculovirus-infected Sf9 cells and tested for their ability to bind a range of fatty acids including unsubstituted (lauric acid (C12:0) and oleic acid (C18:1)) and oxygenated (9,10-epoxystearic acid and 9,10-dihydroxystearic acid). Comparison between these five P450s at constant P450 content over a range of concentrations for individual fatty acids indicates that binding of different fatty acids to CYP86A2 always results in a higher proportion of high spin state heme than binding titrations conducted with CYP86A1 or CYP86A4. In comparison to these three, CYP86A7 and CYP86A8 produce extremely low proportions of high spin state heme even with the most effectively bound fatty acids. In addition to their previously demonstrated lauric acid hydroxylase activities, all CYP86A proteins are capable of hydroxylating oleic acid but not oxygenated 9,10-epoxystearic acid. Homology models have been built for these five enzymes that metabolize unsubstituted fatty acids and sometimes bind oxygenated fatty acids. Comparison of the substrate binding modes and predicted substrate access channels indicate that all use channel pw2a consistent with the crystal structures and models of other fatty acid-metabolizing P450s in bacteria and mammals. Among these P450s, those that bind internally oxygenated fatty acids contain polar residues in their substrate binding cavity that help stabilize these charged/polar groups within their largely hydrophobic catalytic site. PMID- 17427947 TI - beta-sitosterol decreases irradiation-induced thymocyte early damage by regulation of the intracellular redox balance and maintenance of mitochondrial membrane stability. AB - Both radiation injury and oxidation toxicity occur when cells are exposed to ion irradiation (IR), ultimately leading to apoptosis. This study was designed to determine the effect of beta-sitosterol (BSS) on early cellular damage in irradiated thymocytes and a possible mechanism of effect on irradiation-mediated activation of the apoptotic pathways. Thymocytes were irradiated (6 Gy) with or without BSS. Cell apoptosis and apoptosis-related proteins were evaluated. BSS decreased irradiation-induced cell death and nuclear DNA strand breaks while attenuating intracellular reactive oxygen species (ROS) and increasing the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). BSS decreased the release of cytochrome c from mitochondria to the cytosol and the mitochondrio-nuclear translocation of apoptosis-inducing factor (AIF). Furthermore, BSS partially inhibited the radiation-induced increase of cleaved caspase 3 and cleaved PARP, and attenuated the activation of JNK and AP-1. In addition, evidence suggests that ROS generated by irradiation are involved in this course of cell damage. The results indicate that BSS confers a radioprotective effect on thymocytes by regulation of the intracellular redox balance which is carried out via the scavenging of ROS and maintenance of mitochondrial membrane stability. PMID- 17427948 TI - Targeted molecular dynamics simulation studies of binding and conformational changes in E. coli MurD. AB - Enzymes involved in the biosynthesis of bacterial peptidoglycan, an essential cell wall polymer unique to prokaryotic cells, represent a highly interesting target for antibacterial drug design. Structural studies of E. coli MurD, a three domain ATP hydrolysis driven muramyl ligase revealed two inactive open conformations of the enzyme with a distinct C-terminal domain position. It was hypothesized that the rigid body rotation of this domain brings the enzyme to its closed active conformation, a structure, which was also determined experimentally. Targeted molecular dynamics 1 ns-length simulations were performed in order to examine the substrate binding process and gain insight into structural changes in the enzyme that occur during the conformational transitions into the active conformation. The key interactions essential for the conformational transitions and substrate binding were identified. The results of such studies provide an important step toward more powerful exploitation of experimental protein structures in structure-based inhibitor design. PMID- 17427949 TI - Induction of immature dendritic cell apoptosis by foot and mouth disease virus is an integrin receptor mediated event before viral infection. AB - Foot and mouth disease virus (FMDV) has been demonstrated to infect dendritic cells (DC) and reduced its ability to stimulate host immune responses. This study aimed to determine whether non-replicating FMDV could induce apoptosis of the host immune cells. In this study, we have demonstrated that bone morrow derived dendritic cells (BMDCs) were induced to undergo apoptosis in a dose-dependent manner, which was determined by the annexin-V staining, DNA fragmentation, and TUNEL staining methods, after they were treated with the chemically inactivated FMDV in vitro. The initiation of apoptosis was apparently via an interaction of the integrin receptor on BMDCs and the RGD motif within the VP1 capsid protein of FMDV. The initiation activated a cascade of apoptotic pathway including reduced expression of Bcl-2, activation of caspases, and release of cytochrome c from mitochondria. Pretreatment with BMDCs with LPS prevented the inactivated FMDV induced apoptosis, suggesting immature BMDCs are susceptible to such apoptosis. Taken together, the data demonstrate that the inactivated FMDV induces the apoptosis in BMDCs via the integrin receptor and subsequently triggers the apoptosis signal, suggesting that such induction of apoptosis is likely to impair immune responses against FMDV infection. PMID- 17427950 TI - Melatonin reduces protein and lipid oxidative damage induced by homocysteine in rat brain homogenates. AB - Numerous data indicate that hyperhomocysteinemia is a risk factor for cardio- and cerebrovascular diseases. At least in part, homocysteine (HCY) impairs cerebrovascular function because it generates large numbers of free radicals. Since melatonin is a well-known antioxidant, which reduces oxidative stress and decreases HCY concentrations in plasma, the aim of this study was to investigate the effect of melatonin in preventing HCY-induced protein and lipid oxidation in rat brain homogenates. Brain homogenates were obtained from Sprague-Dawley rats and were incubated with or without HCY (0.01-5 mM) or melatonin (0.01-3 mM). Carbonyl content of proteins, and malondialdehyde (MDA) and 4-hydroxyalkenals (4 HDA) concentrations in the brain homogenates were used as an index of protein and lipid oxidation, respectively. Under the experimental conditions used, the addition of HCY (0.01-5 mM) to the homogenates enhanced carbonyl protein and MDA+4-HDA formation. Melatonin reduced, in a concentration-dependent manner, protein and lipid oxidation due to HCY in the brain homogenates. These data suggest that preserving proteins from oxidative insults is an additional mechanism by which melatonin may act as an agent in potentially decreasing cardiovascular and cerebrovascular diseases related to hyperhomocysteinemia. PMID- 17427951 TI - Nanomechanotransduction and interphase nuclear organization influence on genomic control. AB - The ability of cells to alter their genomic regulation in response to mechanical conditioning or through changes in morphology and the organization of the interphase nuclei are key questions in cell biology. Here, two nanotopographies have been used as a model surfaces to change cell morphology in order to investigate spatial genomic changes within the nuclei of fibroblasts. Initially, centromeres for chromosome pairs were labeled and the average distance on different substrates calculated. Further to this, Affymetrix whole genome GeneChips were used to rank genomic changes in response to topography and plot the whereabouts on the chromosomes these changes were occurring. It was seen that as cell spreading was changed, so were the positions along the chromosomes that gene regulations were being observed. We hypothesize that as changes in cell and thus nuclear morphology occur, that this may alter the probability of transcription through opening or closing areas of the chromosomes to transcription factors. PMID- 17427952 TI - A "moving metal mechanism" for substrate cleavage by the DNA repair endonuclease APE-1. AB - Apurinic/apyrimidinic endonuclease (APE-1) is essential for base excision repair (BER) of damaged DNA. Here molecular dynamics (MD) simulations of APE1 complexed with cleaved and uncleaved damaged DNA were used to determine the role and position of the metal ion(s) in the active site before and after DNA cleavage. The simulations started from an energy minimized wild-type structure of the metal free APE1/damaged-DNA complex (1DE8). A grid search with one Mg2+ ion located two low energy clusters of Mg2+ consistent with the experimentally determined metal ion positions. At the start of the longer MD simulations, Mg2+ ions were placed at different positions as seen in the crystal structures and the movement of the ion was followed over the course of the trajectory. Our analysis suggests a "moving metal mechanism" in which one Mg2+ ion moves from the B- (more buried) to the A-site during substrate cleavage. The anticipated inversion of the phosphate oxygens occurs during the in-line cleavage reaction. Experimental results, which show competition between Ca2+ and Mg2+ for catalyzing the reaction, and high concentrations of Mg2+ are inhibitory, indicate that both sites cannot be simultaneously occupied for maximal activity. PMID- 17427953 TI - Crystal structure of human slingshot phosphatase 2. PMID- 17427954 TI - Effect of thermal stress on early and late passaged mouse lens epithelial cells. AB - Cataract is an age related disease of protein aggregation. It has been suggested that aging affects the cells ability to protect protein integrity. The protein integrity, which is essential for cellular homeostasis, is maintained by a complex system of refolding or degradation of damaged proteins. The heat shock proteins (hsps) are the major contributors in the maintenance of protein integrity. The heat shock transcription factor (HSF-1) is the master regulator of all hsp synthesis in response to stress. This investigation examined the role of HSF-1 in the regulation of hsp synthesis in early and late passaged alphaTN-4 cells. Data collected in this study revealed that the nucleotide sequence of HSF 1 mRNA obtained from early and late passaged alphaTN-4 cells were identical. When early and late passaged cell were exposed to thermal stress, their hsp expression were also similar. HSP-40 expression was detected after 2 h of heat stress, whereas HSP-70 and low molecular weight heat shock protein alphabeta crystallin showed significantly increased synthesis 18 h post heat stress. The late passaged alphaTN-4 cells ability to upregulate hsps in response to heat stress could be due to its high replicative activities. The data presented here suggests a relationship between the presence of functional HSF-1 and sustained proliferative activities of the late passaged alphaTN-4 cell. PMID- 17427955 TI - Fusion expression of DDR2 extracellular domain in insect cells and its purification and function characterization. AB - Discoidin domain receptor 2 (DDR2) is a kind of protein tyrosine kinases associated with cell proliferation and tumor metastasis, and collagen, a ligand for DDR2, up-regulates matrix metalloproteinase 1 (MMP-1) and MMP-2 expression in extracellular matrix (ECM). To investigate the role of DDR2 in cartilage destruction in rheumatoid arthritis (RA), we expressed the extracellular domain (ECD) of DDR2 (without signal peptide and transmembrane domain, designated DR) in insect cells, purified and characterized DR, hoping to use it as a specific antagonist of DDR2. By using Bac-To-Bac Expression System with a His tag, we successfully obtained the recombinant bacularvirus containing DDR2 ECD, purified it and characterized its function. The soluble fraction of DR was about 12% of the total fused protein. After chromatographic purification, DR with 92% purity was obtained. Competitive inhibition assay demonstrated that DR blocked the binding between DDR2 and natural DDR2 receptors on NIH3T3 and synovial cells. Results of RT-PCR, Western blotting, and gelatinase zymography showed that DR was capable of inhibiting MMP-1 and MMP-2 secretion from NIH3T3 and RA synoviocytes stimulated by collagen II. For MMP-1, inhibition was displayed at the levels of mRNA and protein, whereas for MMP-2 it was at the level of protein. These findings suggested that the expressed DR inhibited the activity of natural DDR2 and relevant MMP-1 and MMP-2 expression in RA synoviocytes and NIH3T3 cells provoked by collagen II. PMID- 17427956 TI - Pulsed electromagnetic fields rapidly modulate intracellular signaling events in osteoblastic cells: comparison to parathyroid hormone and insulin. AB - Pulsed electromagnetic field (PEMF) devices are approved for the healing of bone nonunions, but there is a lack of understanding as to their mechanism of action at the cell and molecular level. Intermittent parathyroid hormone (PTH) therapy is currently utilized for treatment of osteoporosis, and is also being investigated for the purpose of augmenting fracture healing. Insulin and IGF-1 are also thought to play important anabolic roles in osteogenesis. In this report, signaling pathways activated by acute PTH or insulin treatments were compared to those activated by PEMF treatment in osteoblast-like cells. Some signaling molecules like the extracellular response kinases 1/2 (Erk1/2) and the cAMP response element binding protein (CREB) were activated by insulin and PTH, respectively, but not by PEMF treatment. Other signaling molecules like the insulin receptor substrate-1 (IRS-1), the S6 ribosomal subunit kinase, and the endothelial nitric oxide synthase (eNOS) were phosphorylated by PTH, insulin, and PEMF to the same relative extent and within the same time frame. IRS-1, eNOS, and S6 have been implicated in bone anabolism, and our results suggest that the anabolic effects of PEMF may be mediated, in part, through the activation of these proteins. PMID- 17427957 TI - Accuracy of binding mode prediction with a cascadic stochastic tunneling method. AB - We investigate the accuracy of the binding modes predicted for 83 complexes of the high-resolution subset of the ASTEX/CCDC receptor-ligand database using the atomistic FlexScreen approach with a simple forcefield-based scoring function. The median RMS deviation between experimental and predicted binding mode was just 0.83 A. Over 80% of the ligands dock within 2 A of the experimental binding mode, for 60 complexes the docking protocol locates the correct binding mode in all of ten independent simulations. Most docking failures arise because (a) the experimental structure clashed in our forcefield and is thus unattainable in the docking process or (b) because the ligand is stabilized by crystal water. PMID- 17427958 TI - Structural and kinetic analysis of proton shuttle residues in the active site of human carbonic anhydrase III. AB - We report the X-ray crystal structures and rate constants for proton transfer in site-specific mutants of human carbonic anhydrase III (HCA III) that place a histidine residue in the active-site cavity: K64H, R67H, and K64H-R67N HCA III. Prior evidence from the exchange of 18O between CO2 and water measured by mass spectrometry shows each mutant to have enhanced proton transfer in catalysis compared with wild-type HCA III. However, His64 in K64H and K64H-R67N HCA III have at most a capacity for proton transfer that is only 13% that of His64 in HCA II. This reduced rate in mutants of HCA III is associated with a constrained side chain conformation of His64, which is oriented outward, away from the active-site zinc in the crystal structures. This conformation appears stabilized by a prominent pi stacking interaction of the imidazole ring of His64 with the indole ring of Trp5 in mutants of HCA III. This single orientation of His64 in K64H HCA III predominates also in a double mutant K64H-R67N HCA III, indicating that the positive charge of Arg67 does not influence the observed conformation of His64 in the crystal structure. Hence, the structures and catalytic activity of these mutants of HCA III containing His64 account only in small part for the lower activity of this isozyme compared with HCA II. His67 in R67H HCA III was also shown to be a proton shuttle residue, having a capacity for proton transfer that was approximately four times that of His64 in K64H HCA III. This is most likely due to its proximity and orientation inward towards the zinc-bound solvent. These results emphasize the significance of side chain orientation and range of available conformational states as characteristics of an efficient proton shuttle in carbonic anhydrase. PMID- 17427959 TI - The transcription factor SOX17 is involved in the transcriptional control of the uteroglobin gene in rabbit endometrium. AB - The transcription of the uteroglobin gene (ug) is induced by progesterone in the rabbit endometrium, primarily through the binding of the progesterone receptor to the distal region of the ug promoter. However, other transcription factors participate in the progesterone action. The proximal ug promoter contains several putative consensus sequences for the binding of various progesterone-dependent endometrial nuclear factors (Perez Martinez et al. [1996] Arch Biochem Biophys 333: 12-18), suggesting that several transcription factors might be implicated in the hormonal induction of ug. We report here that one of these progesterone dependent factors specifically binds to the sequence CACAATG (-183/-177) of the rabbit ug promoter. This sequence (hereafter called element G') is very similar to the consensus sequence for binding of the SOX family of transcription factors. Mutation of the element G' reduced transcription from the ug promoter in transient expression experiments. The endometrial factor was purified and analyzed by nano-liquid chromatography and ion trap coupled mass spectrometry yielding two partial amino acid sequences corresponding to a region of SOX17 that is highly conserved inter-species. This identification was confirmed by immunological techniques using a specific anti-SOX17 antibody. In agreement with the above findings, overexpression of SOX17 in transfected endometrial cells increased transcription from the ug promoter. SOX17 gradually accumulated in the nucleus in vivo concomitant with the induction of ug expression by progesterone in the endometrium. Thus, these findings implicate, for the first time, SOX17 in the transcriptional control of rabbit ug. PMID- 17427960 TI - Inhibition of GSK-3beta decreases NF-kappaB-dependent gene expression and impairs the rat liver regeneration. AB - Serine-threonine protein kinase glycogen synthase kinase (GSK)-3 is involved in regulation of many cell functions, but its role in regulating liver regeneration is unknown. Here we investigated the effects of GSK-3beta inhibition on liver regeneration after partial hepatectomy in the rat. The potent and selective GSK 3beta inhibitor SB216763 (0.6 mg/kg intravenously) or vehicle (10% dimethyl sulfoxide) was administered 30 min before 70% partial hepatectomy. Liver regeneration was estimated by the cell proliferation, apoptosis, and the related cell signaling and cycling proteins. In 30 min after hepatectomy in the rat, GSK 3beta was found to be translocated to the nucleus, but GSK-3beta inhibitor SB216763 that could phosphorylate residue Ser9 on GSK-3beta did not attenuated the accumulation. Consequently, the inhibition of GSK-3beta decreased the nuclear factor-kappaB activity, the NF-kappaB-dependent gene expression, and COX2 expression, but enhanced p21(WAF1/Cip1) transcription. Moreover, the injection of SB216763 impaired the proliferation cell nuclear antigen (PCNA) index and increased the apoptosis of liver compared to the vehicle. GSK-3beta plays an important role in rat liver regeneration. We conclude it may partially result from the inhibition of the NF-kappaB pathway and enhancement of p21 (WAF1/Cip1) expression. PMID- 17427961 TI - Structural prediction of the beta-domain of metallothionein-3 by molecular dynamics simulation. AB - The beta-domain of metallothionein-3 (MT3) has been reported to be crucial to the neuron growth inhibitory bioactivity. Little detailed three-dimensional structural information is available to present a reliable basis for elucidation on structure-property-function relationships of this unique protein by experimental techniques. So, molecular dynamics simulation is adopted to study the structure of beta-domain of MT3. In this article, a 3D structural model of beta-domain of MT3 was generated. The molecular simulations provide detailed protein structural information of MT3. As compared with MT2, we found a characteristic conformation formed in the fragment (residue 1-13) at the N terminus of MT3 owing to the constraint induced by 5TCPCP9, in which Pro7 and Pro9 residues are on the same side of the protein, both facing outward and the two 5-member rings of prolines are arranged almost in parallel, while Thr5 is on the opposite side. Thr5 in MT3 is also found to make the first four residues relatively far from the fragment (residue 23-26) as compared with MT2. The simulated structure of beta-domain of MT3 is looser than that of MT2. The higher energy of MT3 than that of MT2 calculated supports these conclusions. Simulation on the four isomer arising from the cis- or trans-configuration of 6CPCP9 show that the trans-/trans-isomer is energetic favorable. The partially unfolding structure of beta-domain of MT3 is also simulated and the results show the influence of 6CPCP9 sequence on the correct folding of this domain. The correlations between the bioactivity of MT3 and the simulated structure as well as the folding of beta-domain of MT3 are discussed based on our simulation and previous results. PMID- 17427962 TI - Prostaglandin E2 induces vascular endothelial growth factor secretion in prostate cancer cells through EP2 receptor-mediated cAMP pathway. AB - Prostaglandin E2 (PGE2) has been shown to induce expression of vascular endothelial growth factor (VEGF) and other signaling molecules in several cancers. PGE2 elicits its functions though four G-protein coupled membrane receptors (EP1-4). In this study, we investigated the role of EP receptors in PGE2-induced molecular events in prostate cancer cells. qRT-PCR analysis revealed that PC-3 cells express a substantially higher level of EP2 and moderately higher EP4 than DU145 and LNCaP cells. LNCaP cells had virtually no detectable EP2 mRNA. EP1 and EP3 mRNAs were not detected in these cells. Treatment of prostate cancer cells with PGE2 (1 nM-10 microM) increased both VEGF secretion and cyclic adenosine monophosphate (cAMP) production. Levels of induction in PC-3 cells were greater than in DU145 and LNCaP cells. The selective EP2 agonist CAY10399 also significantly increased VEGF secretion and cAMP production in PC-3 cells, but not in DU145 and LNCaP cells. Moreover, PGE2 and CAY10399 increased mitogen activated protein kinase/extracellular signal regulated kinase (MAPK/Erk) and Akt phosphorylation in PC-3 and DU145 cells, but not in LNCaP cells. However, neither the MAPK/Erk inhibitor U0126 nor the PI3K/Akt inhibitor LY294002 abolished PGE2 induced VEGF secretion in PC-3 cells. We further demonstrated that the adenylate cyclase activator forskolin and the cAMP anologue 8-bromo-cAMP mimicked the effects of PGE2 on VEGF secretion in PC-3 cells. Meanwhile, the adenylate cyclase inhibitor 2'5'-dideoxyadenosine, at concentrations that inhibited PGE2-induced cAMP, significantly blocked PGE2-induced VEGF secretion in PC-3 cells. We conclude that PGE2-induced VEGF secretion in prostate cancer cells is mediated through EP2-, and possibly EP4-, dependent cAMP signaling pathways. PMID- 17427963 TI - Birth of cloned miniature pigs derived from somatic cell nuclear transferred embryos activated by ultrasound treatment. AB - The present study was carried out to determine (1) the optimal duty cycle of ultrasound for activation of pig oocytes and cloned embryos derived from miniature pig fetal fibroblasts and (2) whether cloned embryos can develop to term following activation by ultrasound stimulation. When oocytes were exposed to ultrasound with 20% or 30% duty cycle, the blastocyst formation rates were significantly (P < 0.05) higher than that of oocytes exposed to ultrasound with 10% duty cycle. In contrast, the blastocyst formation rate of cloned embryos decreased as the duty cycle of ultrasound increased; the value of embryos exposed to ultrasound with 10% duty cycle was significantly (P < 0.05) higher than that of embryos exposed to ultrasound with 50% duty cycle. When cloned embryos exposed to ultrasound with 10% duty cycle were transferred into the oviducts of two recipient gilts to assess their development in vivo, the pregnancy of one of the gilts was maintained to term and two piglets were delivered via Cesarean section. The results of the present study showed that (1) although the duty cycle of ultrasound affects in vitro development after activation of both pig oocytes and miniature pig cloned embryos, the optimal duty cycle is different between them and (2) miniature pig cloned embryos have the ability to develop into piglets after activation by ultrasound stimulation. PMID- 17427964 TI - Surgery for head and neck skin tumors in the elderly. AB - BACKGROUND: Our aim was to determine the surgical characteristics of an elderly cohort of patients undergoing resection of head and neck skin neoplasms. METHODS.: All cases of patients with head and neck skin neoplasms (excluding malignant melanoma) who underwent surgical resection in a regional referral center over a 10-year period were retrospectively reviewed. The study group comprised 152 patients (208 cases) aged >or=80 years. They were compared with 311 patients (430 cases) aged <80 years. RESULTS: Elderly patients had higher Charlson comorbidity index, American Society of Anesthesiologists' physical status scores, dementia, and aspirin intake (all p < .05). They also had a higher incidence of scalp involvement and lower incidence of nasal involvement (both p < .05). Elderly patients had a larger lesion size at presentation and required sacrifice of a greater area of skin (both p < .05). However, they also underwent more local anesthetic procedures, although the need for local flap or skin graft repair was not increased. Two elderly patients had intraoperative tachyarrythmias induced by local anesthesia. However, no patient died within 30 days of surgery, and wound complication rate and disease-free survival were comparable to younger patients. CONCLUSION: With careful attention to comorbid illness and perioperative monitoring, surgical resection of head and neck skin neoplasms is safe in the elderly. Lesions are more advanced at presentation and hence require sacrifice of a larger area of skin to obtain macroscopic clearance. Yet for the majority of lesions, local anesthesia is adequate and surgical resection and simple skin closure can be accomplished without the need for complex flap or skin graft reconstructions. PMID- 17427965 TI - Ruptured pseudoaneurysm of the internal maxillary artery complicating CT-guided fine-needle aspiration in an irradiated, surgical bed. AB - BACKGROUND: CT-guided fine-needle aspiration (FNA) is a safe procedure, but major complications can occur rarely. Pseudoaneurysm rupture in the head and neck region following CT-guided FNA is an emergency that can result in life threatening hemorrhage. This case emphasizes the salient risk factors for pseudoaneurysm formation and rupture in the head and neck region following CT guided FNA. METHODS: A patient was seen with oral and facial hemorrhage as a result of a ruptured pseudoaneurysm 11 weeks following CT-guided FNA in a previously irradiated surgical bed. RESULTS: The patient was treated with coil embolization in and around the pseudoaneurysm and discharged without any further complications. CONCLUSIONS: Although CT-guided FNA is a safe and effective procedure, some patients may be at increased risk for rare but major complications. Caution should be used in proceeding with CT-guided FNA in an irradiated surgical bed of the head and neck. PMID- 17427966 TI - Endonasal endoscopic resection and radiotherapy in olfactory neuroblastoma: our experience. AB - BACKGROUND: Our aim was to evaluate the efficacy of a bimodal method of treatment consisting in endoscopic resection followed by radiotherapy in patients with olfactory neuroblastoma (ON). METHODS: This is a retrospective review on 10 patients with ON treated at a tertiary referral center. All the patients were treated with endonasal endoscopic resection, and 1 refused postoperative radiotherapy. RESULTS: No mortality was observed. Local tumor control was obtained in all the patients. Follow-up ranged from 15 to 79 months (median, 37 months). One patient developed a regional recurrence and for this was treated with bilateral, modified type III radical neck dissection plus radiotherapy on the neck. All patients regained a good quality of life after the treatment. CONCLUSIONS: This method of treatment causes minimal injury to the patients, reduces side effects, and improves the quality of life of those with olfactory neuroblastoma. PMID- 17427967 TI - Cervical lipoblastoma: case report, review of literature, and genetic analysis. AB - BACKGROUND: Lipoblastoma is a rare, benign tumor of infants and children, usually occurring in the extremities and trunk, with only a few cases reported in the neck. METHODS: We describe the case of an infant with a rapidly enlarging, painless neck mass. MRI revealed a 4-cm-diameter mass deep to the paraspinal muscles, in close proximity to the C2 vertebral foramen. Review of literature, diagnostic methods, and genetics of lipomatous tumors are discussed. RESULTS: Complete surgical excision via a posterior cervical approach demonstrated irregular lobules of immature fat cells separated by a loose, myxoid connective tissue. Histology and genetic analysis confirmed the diagnosis of lipoblastoma. CONCLUSION: Cervical lipoblastoma is rare, and typically presents as an asymptomatic, painless mass, rarely causing airway obstruction or nerve compression. MRI can be helpful in identifying the lipomatous nature of the mass, but the findings can be inconsistent due to variable maturity of fat cells and the mesenchymal content of the tumor. Chromosomal analysis is useful in differentiating lipoblastoma from liposarcoma. Recommended treatment is complete surgical excision. PMID- 17427968 TI - Clinical and operative management of persistent hyperparathyroidism after renal transplantation: a single-center experience. AB - BACKGROUND: Persistent (tertiary) hyperparathyroidism (TH) after renal transplantation may cause considerable morbidity and necessitate parathyroidectomy. This study investigated the characteristics of this patient subgroup. METHODS: The medical data and pathology specimens of 20 kidney transplant recipients who underwent parathyroidectomy for TH in 2001 to 2004 were reviewed. RESULTS: Treatment consisted of subtotal resection of 3.5 glands in 13 patients, resection of 3 to 3.5 glands under intraoperative parathyroid hormone monitoring (iPTH) in 5 patients, and selective resection in 2 patients with markedly asymmetric gland enlargement. Eighteen patients had hyperplasia-diffuse in 10, nodular in 4, or both in 2; 2 patients had 1 large nodule in every gland. Six patients had postoperative complications. Follow-up of 2 years revealed recurrent hypercalcemia in 1 patient and a high level of PTH (>60 pg/mL) in 12. CONCLUSION: Subtotal resection for TH may be insufficient. The use of iPTH monitoring is recommended. Renal transplant recipients have distinctive characteristics and require special perioperative attention. PMID- 17427969 TI - Locoregional control in patients with palpable medullary thyroid cancer: results of standardized compartment-oriented surgery. AB - BACKGROUND: Extent of neck dissection is controversial in patients with palpable medullary thyroid cancer (MTC). METHODS: We evaluated 64 MTC patients (19 hereditary, 45 sporadic) with palpable thyroid nodules (group 1, n = 35) or palpable lymph node metastases (group 2, n = 29). Standard surgery included total thyroidectomy, central compartment dissection, and additional neck dissection on indication. RESULTS: In group 1, 40% of the patients were cured. Thirty-one percent of all patients had central, 23% ipsilateral, 14% contralateral, and 14% mediastinal, metastases. Fifty-one percent developed locoregional recurrence. Locoregional recurrence (p = .043) and reoperations (p = .020) were noted more often after a less than standard initial procedure. In group 2, no patients were cured. All had central, 93% ipsilateral, 45% contralateral, and 52% mediastinal metastases. Thirty-eight percent developed locoregional recurrence. CONCLUSIONS: Locoregional recurrence frequently occurs in palpable MTC, and tumor control may be improved by standard central, bilateral, and upper mediastinal neck dissection. PMID- 17427970 TI - Mycobacterium fortuitum-induced persistent parotitis: successful therapy with clarithromycin and ciprofloxacin. AB - BACKGROUND: Parotitis caused by nontuberculous mycobacteria, a very rare disease entity, has never been reported to be caused by Mycobacterium fortuitum (M. fortuitum) in the literature. METHODS AND RESULTS: An 8-year-old girl was seen with painful swelling of the right parotid gland despite antibiotic treatment of more than 1 month. Elevated serum amylase activity and diffuse contrast-enhanced CT of the parotid gland confirmed the diagnosis of parotitis. Histopathological study of specimens taken from the right parotid tail mass showed granulomatous inflammation with acid-fast positive bacilli; culture later confirmed M. fortuitum. After administration of clarithromycin and ciprofloxacin for 9 consecutive months, the parotitis and parotid tail mass were completely resolved at follow-up examination. CONCLUSION: To our knowledge, this is the first case report of parotitis caused by M. fortuitum and its successful medical treatment. PMID- 17427971 TI - Prognostic significance of cyclooxygenase-2 and DNA topoisomerase IIalpha expression in oral carcinoma. AB - BACKGROUND: Despite recent advances in the diagnosis and treatment of oral carcinoma, outcomes remain disappointing. The identification of new prognostic factors is necessary to improve survival. To determine the prognostic significance of cyclooxygenase (COX)-2 and DNA topoisomerase (DNA-Topo) IIalpha expression in patients with oral carcinoma, we immunohistochemically examined these enzymes and studied their relation to overall 5-year survival. METHODS: Surgical specimens were obtained from 160 patients with oral carcinoma, 80 with and 80 without regional lymph node metastasis. The specimens were immunostained for COX-2 and DNA-Topo IIalpha as an index of cell proliferative activity. COX-2 immunoreactivity and clinicopathological data were analyzed, and 5-year survival was calculated by the Kaplan-Meier method. RESULTS: COX-2 expression in primary lesions was higher in cases with lymph node metastasis than in those without lymph node metastasis. An increase in tumor size was associated with increased COX-2 expression. In most cases with lymph node metastasis, COX-2 expression was higher in metastatic lesions than in primary lesions. As COX-2 expression increased, the DNA-Topo IIalpha labeling index significantly increased and the overall 5-year survival rate decreased. CONCLUSION: Expression of COX-2 and DNA Topo IIalpha were related to lymph node metastasis, cell proliferative activity, and overall 5-year survival rate in oral carcinoma. These enzymes may therefore be valuable diagnostic and prognostic indices in oral carcinoma. PMID- 17427972 TI - Problems in the current TNM staging of nonmelanoma skin cancer of the head and neck. PMID- 17427973 TI - Experimental metamorphosis of Halisarca dujardini larvae (Demospongiae, Halisarcida): evidence of flagellated cell totipotentiality. AB - The potency of flagellated cells of Halisarca dujardini (Halisarcida, Demospongiae) larvae from the White Sea (Arctic) was investigated experimentally during metamorphosis. Two types of experiments were conducted. First, larvae were maintained in Ca2+ free seawater (CFSW) until the internal cells were released outside through the opening of the posterior pole. These larvae that only composed of flagellated cells (epithelial larvae) were then returned to sea water (SW) to observe their metamorphosis. The posterior aperture closed before they settled on a substratum and started a metamorphosis similar to intact larvae. Secondly, epithelial larvae were, first, further treated in CFSW and then mechanically dissociated. Separated cells or groups of cells were returned to SW, where they constituted large friable conglomerates. After 12-17 h in SW, flagellated cells showed the first steps of dedifferentiation, and regional differentiation was noticeable within conglomerates after approximately 24-36 h. External cells differentiated into pinacocytes while internal cells kept their flagella and became united in a layer. Within 48-72 h, internal cells of the conglomerates formed spherical or ovoid clusters with an internal cavity bearing flagella. These clusters further fused together in a rhagon containing one or two large choanocyte chambers. The sequence of cellular processes in epithelial larvae and in flagellated cell conglomerates was similar. Previous observations indicating the totipotentiality of larval flagellated cells during normal metamorphosis of H. dujardini are thus confirmed. PMID- 17427974 TI - Metamorphosis of cinctoblastula larvae (Homoscleromorpha, porifera). AB - The metamorphosis of the cinctoblastula of Homoscleromorpha is studied in five species belonging to three genera. The different steps of metamorphosis are similar in all species. The metamorphosis occurs by the invagination and involution of either the anterior epithelium or the posterior epithelium of the larva. During metamorphosis, morphogenetic polymorphism was observed, which has an individual character and does not depend on either external or species specific factors. In the rhagon, the development of the aquiferous system occurs only by epithelial morphogenesis and subsequent differentiation of cells. Mesohylar cells derive from flagellated cells after ingression. The formation of pinacoderm and choanoderm occurs by the differentiation of the larval flagellated epithelium. This is possibly due to the conservation of cell junctions in the external surface of the larval flagellated cells and of the basement membrane in their internal surface. The main difference in homoscleromorph metamorphosis compared with Demospongiae is the persistence of the flagellated epithelium throughout this process and even in the adult since exo- and endopinacoderm remain flagellated. The antero-posterior axis of the larva corresponds to the baso-apical axis of the adult in Homoscleromorpha. PMID- 17427975 TI - NSSR1 is regulated in testes development and cryptorchidism and promotes the exon 5-included splicing of CREB transcripts. AB - Neural salient serine/arginine rich protein 1 (NSSR1, alternatively SRp38) is a newly identified splicing factor that is highly expressed in neural and reproductive tissues. We showed that the expression of testicular NSSR1 increased significantly during mouse testes development. NSSR1 was mainly expressed in germ cells, but barely detected in Sertoli cells. Testicular NSSR1 was mostly phosphorylated and cytosolic in germ cells. In comparison, pituitary NSSR1 was mostly dephosphorylated and nuclear. In the cryptorchid testes, the dephosphorylated NSSR1 was significantly increased. RT-PCR analysis demonstrated that the alternative splicing of CREB and CREM genes was altered in the cryptorchid testes. In addition, CREB transcripts were associated with NSSR1 either in testes tissues or cultured GC-1 cells. Moreover, the studies with NSSR1 over-expression or silence demonstrated that NSSR1 promoted the exon 5 inclusion of CREB, indicating that NSSR1 is a new factor that regulates the alternative exon 5 inclusion of CREB transcripts. The findings for the first time provide the evidence indicating the potential importance of NSSR1 in testes development, spermatogenesis and cryptorchidism. PMID- 17427976 TI - Variation in fecal testosterone levels, inter-male aggression, dominance rank and age during mating and post-mating periods in wild adult male ring-tailed lemurs (Lemur catta). AB - In primate species exhibiting seasonal reproduction, patterns of testosterone excretion in adult males are variable: in some species, peaks correlate with female receptivity periods and heightened male-male aggression over access to estrous females, in others, neither heightened aggression nor marked elevations in testosterone have been noted. In this study, we examined mean fecal testosterone ( f T) levels and intermale aggression in wild adult male ring tailed lemurs residing in three groups at Beza Mahafaly Reserve, Madagascar. Results obtained from mating and post-mating season 2003 were compared to test Wingfield et al. [1990. Am Nat 136:829-846] "challenge hypothesis", which predicts a strong positive relationship between male testosterone levels and male male competition for access to receptive females during breeding season. f T levels and rates of intermale aggression were significantly higher during mating season compared to the post-mating period. Mean f T levels and aggression rates were also higher in the first half of the mating season compared with the second half. Number of males in a group affected rates of intermale agonism, but not mean f T levels. The highest-ranking males in two of the groups exhibited higher mean f T levels than did lower-ranking males, and young males exhibited lower f T levels compared to prime-aged and old males. In the post-mating period, mean male f T levels did not differ between groups, nor were there rank or age effects. Thus, although male testosterone levels rose in relation to mating and heightened male-male aggression, f T levels fell to baseline breeding levels shortly after the early mating period, and to baseline non-breeding levels immediately after mating season had ended, offsetting the high cost of maintaining both high testosterone and high levels of male-male aggression in the early breeding period. PMID- 17427977 TI - Observations of daytime births in two groups of red-handed howlers (Alouatta belzebul) on an island in the Tucurui reservoir in eastern Brazilian Amazonia. AB - Births were observed in two free-ranging groups of red-handed howlers (Alouatta belzebul) in eastern Brazilian Amazonia. Both events occurred in the early afternoon, and were relatively rapid and discrete. No other group members approached the females during parturition, nor attempted to interfere with the neonate in any way. Only one of the mothers ingested the placenta. This same female carried the neonate ventrally during the first month of life and then it was carried dorsally. The other infant was carried by its mother in a dorsal position from birth onward. Both infants survived their first month, but subsequently disappeared, in the second and fourth months of life. The precise causes of their death are unknown. In one case, we speculate that the infant's death was due to infestation by botfly larvae. PMID- 17427979 TI - Evidence-based sepsis therapy: a hospitalist perspective. PMID- 17427981 TI - What will board certification be-and mean-for hospitalists? PMID- 17427982 TI - Missing the forest for the trees. PMID- 17427983 TI - Extreme presentation of bony metastatic disease. PMID- 17427984 TI - Perspectives from the other side of the ocean: a Croatian hospital experience. PMID- 17427985 TI - Discontinuity of care: further thoughts on standardized processes. PMID- 17427993 TI - Affinity precipitation an option for early capture in bioprocessing. AB - Affinity precipitation is a bioseparation technique where the affinity ligand is coupled to a stimuliresponsive polymer. Stimuli-responsive polymers show abrupt, yet reversible, phase transition (precipitation) in response to a small change in an environmental parameter. The corresponding ligand conjugates can be used to co precipitate and thereby capture and isolate target molecules from complex solutions such as culture supernatants and cell lysates. The approach is compatible with a 'discardibles only' type of downstream process and can be scaled over several orders of magnitude. This report discusses the set-up and development of affinity precipitation procedures, the related instrumentation and scale up, as well as applications for the isolation of proteins and polynucleotides. PMID- 17427994 TI - Biotechnological production of xylitol in a three-phase fluidized bed bioreactor with immobilized yeast cells in Ca-alginate beads. AB - Cells of Candida guilliermondii entrapped in Ca-alginate beads were used for xylitol production, from concentrated hemicellulose hydrolyzate of sugarcane bagasse, in a fluidized bed bioreactor (FBR). The maximum xylitol concentration 28.9 g xylitol/L was obtained at a high aeration rate of 600 mL/min after 70 h of fermentation, indicating that the use of high aeration rate in this system is favored for better oxygen transfer into the immobilized cells. The specific xylitol productivity and the xylitol yield were of 0.4 g xylitol/L.h and 0.58 g xylitol/g xylose respectively. The immobilization efficiency at the end of the fermentation was of 65 %. After 90 h of fermentation xylitol productivity and yield decreased to 0.25 g xylitol/L.h and 0.47 g xylitol/g xylose respectively, indicating the beginning of xylitol consumption by the yeast. The use of FBR system with immobilized cells presented high xylitol yield and productivity. PMID- 17427995 TI - Genetic fidelity of long-term micropropagated shoot cultures of vanilla (Vanilla planifolia Andrews) as assessed by molecular markers. AB - Occurrence of genetic variants during micropropagation is occasionally encountered when the cultures are maintained in vitro for long period. Therefore, the micropropagated multiple shoots of Vanilla planifolia Andrews developed from axillary bud explants established 10 years ago were used to determine somaclonal variation using random amplified polymorphic DNA (RAPD) and intersimple sequence repeats markers (ISSR). One thousand micro-plants were established in soil of which 95 plantlets (consisting of four phenotypes) along with the mother plant were subjected to genetic analyses using RAPD and ISSR markers. Out of the 45 RAPD and 20 ISSR primers screened, 30 RAPD and 7 ISSR primers showed 317 clear, distinct and reproducible band classes resulting in a total of 30 115 bands. However, no difference was observed in banding patterns of any of the samples for a particular primer, indicating the absence of variation among the micropropagated plants. Our results allow us to conclude that the micropropagation protocol that we have used for in vitro proliferation of vanilla plantlets for the last 10 years might be applicable for the production of clonal plants over a considerable period of time. PMID- 17427996 TI - Comparison of quasisteady-state performance of the DEAMOX process under intermittent and continuous feeding and different nitrogen loading rates. AB - The recently developed denitrifying ammonium oxidation (DEAMOX) process combines the anammox reaction with autotrophic denitrifying conditions using sulfide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. This paper compares a quasisteady-state performance of this process for treatment of baker's yeast wastewater under intermittent and continuous feeding and increasing nitrogen loading rate (NLR) from 300 till 858 mg N/L/d. The average total nitrogen removal slightly decreased on increasing the NLR: from 86 to 79% (intermittent feeding) and from 87 to 84% (continuous feeding). The better performance under continuous feeding was due to a more complete nitrate removal in the former case whereas the ammonia removal was similar for both feeding regimes under the comparable NLR. A possible explanation can be that, during continuous feeding (simultaneous supply of nitrate and sulfide), there were less mass transfer limitations for sulfide oxidizing denitrifiers presumably located in the outer layer of sludge aggregates. On the contrary, the ammonia oxidisers presumably located inside the aggregates apparently suffered from nitrite mass transfer limitations under both the feedings. The paper further describes some characteristics of the DEAMOX sludge. PMID- 17427997 TI - In vivo glyco-engineered antibody with improved lytic potential produced by an innovative non-mammalian expression system. AB - Recent studies have demonstrated that the reduction of the core fucosylation on N glycans of human IgGs is responsible for a clearly enhanced antibody-dependent cellular cytotoxicity (ADCC). This finding might give access to improved active therapeutic antibodies. Here, the expression of the tumor antigen-specific antibody IGN311 was performed in a glyco-optimized strain of the moss Physcomitrella patens. Removal of plant specific N-glycan structures in this plant expression host was achieved by targeted knockout of corresponding genes and included quantitative elimination of core fucosylation. Antibodies transiently expressed and secreted by such genetically modified moss protoplasts assembled correctly, showed an unaltered antigen-binding affinity and, in extensive tests, revealed an up to 40-fold enhanced ADCC. Thus, the glyco engineered moss-based transient expression platform combines a rapid technology with the subsequent analysis of glycooptimized therapeutics with regard to advanced properties. PMID- 17427998 TI - Chemically defined medium environment for the development of renal stem cells into tubules. AB - The use of stem cells is a valuable therapeutical option for the regeneration of diseased tissues and organs. However, the involved cellular processes are hardly known. To gain detailed information about their development, a new culture technology was developed. Embryonic renal tissue containing stem/progenitor cells was mounted within a perfusion culture container at the interface of an artificial interstitium made of polyester. Using this innovative approach we show that renal tubules develop in chemically defined Iscove's modified Dulbecco's medium without serum addition and without coating by extracellular matrix proteins. The development of tubules depends on the administration of aldosterone, and can be visualized by immunohistochemical labeling. The presented technology makes the exact analysis of developmental steps now possible, and provides a new powerful tool to optimize growth and differentiation of renal stem cells. It may also enable many other kinds of stem cells to steer their development into functional tissues under clearly defined in vitro conditions. PMID- 17427999 TI - Determination of cefquinome in pig plasma and bronchoalveolar lavage fluid by high-performance liquid chromatography combined with electrospray ionization mass spectrometry. AB - The aim of this study was to develop a rapid and sensitive method for the quantification of cefquinome in animal plasma and bronchoalveolar lavage (BAL) fluid using high-performance liquid chromatography combined with electrospray tandem mass spectrometry (LC-ESI-MS/MS). Cefadroxil is used as internal standard. For plasma, the sample preparation includes a simple deproteinization step with a Microcon filter. This allows detecting the unbound cefquinome concentration, which is correlated with the concentration in other body fluids, such as BAL fluid. To be able to detect the total plasma concentration, deproteinization with acetonitrile, followed by a back-extraction of actonitrile with dichloromethane was performed. The BAL fluid is centrifuged to precipitate floating particles. Chromatographic separation is achieved on a PLRP-S column using 0.005% formic acid and methanol as mobile phase. For plasma, good linearity was observed in the range of 5-2500 ng ml(-1) for both the unbound and total concentration. The response in BAL fluid was linear in the range of 4-1000 ng ml(-1). The limit of quantification (LOQ) was set at 5.00 ng ml(-1) for plasma and at 4.00 ng ml(-1) for BAL fluid. The limit of detection (LOD) was 3.12 ng ml(-1) and 0.41 ng ml(-1) for the unbound and total concentration in plasma, respectively, and was 1.43 ng ml(-1) for BAL fluid. The method was shown to be of use in a pharmacokinetic study in pigs, where the correlation between cefquinome concentrations in plasma and BAL fluid of pigs was studied. PMID- 17428000 TI - Characterization of N-palmitoylated human growth hormone by in situ liquid-liquid extraction and MALDI tandem mass spectrometry. AB - Acylation is a common post-translational modification found in secreted proteins and membrane-associated proteins, including signal transducing and regulatory proteins. Acylation is also explored in the pharmaceutical and biotechnology industry to increase the stability and lifetime of protein-based products. The presence of acyl moieties in proteins and peptides affects the physico-chemical properties of these species, thereby modulating protein stability, function, localization and molecular interactions. Characterization of protein acylation is a challenging analytical task, which includes the precise definition of the acylation sites in proteins and determination of the identity and molecular heterogeneity of the acyl moiety at each individual site. In this study, we generated a chemically modified human growth hormone (hGH) by incorporation of a palmitoyl moiety on the N(epsilon) group of a lysine residue. Monoacylation of the hGH protein was confirmed by determination of the intact molecular weight by mass spectrometry. Detailed analysis of protein acylation was achieved by analysis of peptides derived from hGH by protease treatment. However, peptide mass mapping by MALDI MS using trypsin and AspN proteases and standard sample preparation methods did not reveal any palmitoylated peptides. In contrast, in situ liquid-liquid extraction (LLE) performed directly on the MALDI MS metal target enabled detection of acylated peptide candidates by MALDI MS and demonstrated that hGH was N-palmitoylated at multiple lysine residues. MALDI MS and MS/MS analysis of the modified peptides mapped the N-palmitoylation sites to Lys158, Lys172 and Lys140 or Lys145. This study demonstrates the utility of LLE/MALDI MS/MS for mapping and characterization of acylation sites in proteins and peptides and the importance of optimizing sample preparation methods for mass spectrometry-based determination of substoichiometric, multi-site protein modifications. PMID- 17428001 TI - Localization of CiCBR in the invertebrate chordate Ciona intestinalis: evidence of an ancient role for cannabinoid receptors as axonal regulators of neuronal signalling. AB - CiCBR is a G-protein-coupled receptor in the sea-squirt Ciona intestinalis and the first ortholog of vertebrate CB(1) and CB(2) cannabinoid receptors to be identified in an invertebrate (Elphick et al. [2003] Gene 302:95-101). Here we have used Western blotting and immunocytochemistry to examine expression of CiCBR in adult Ciona, employing novel antibodies to the C-terminal tail of CiCBR. Consistent with the expected mass for CiCBR, a approximately 47-kDa band was detected in Ciona membranes, and immunocytochemical analysis of serial sections of Ciona revealed intense immunoreactivity in the cerebral ganglion localised in a dense meshwork of fibers in the neuropile. Accordingly, Western blot analysis of neural complex homogenates revealed the presence of a approximately 47-kDa band. CiCBR immunoreactivity was also observed in axons exiting the ganglion in the anterior and posterior nerves, and analysis of whole-mount preparations revealed that these axons project over the interior surface of the oral and atrial siphons. Isolated CiCBR-immunoreactive axons not associated with the anterior and posterior nerves were observed projecting through the cortical layer of the cerebral ganglion. Central and peripheral CiCBR-immunoreactive fibers were studded with intensely stained varicosities, indicative of a role for CiCBR in regulation of axonal release of neurotransmitters, neuromodulators, or neurohormones. Collectively, our data suggest that the well-established role that the CB(1) receptor has as an axonal regulator of neurotransmitter release in mammals may have originated with ancestral-type cannabinoid receptors in invertebrate chordates before the emergence of CB(1)- and CB(2)-type receptors in vertebrates. PMID- 17428002 TI - Different levels of CD52 antigen expression evaluated by quantitative fluorescence cytometry are detected on B-lymphocytes, CD 34+ cells and tumor cells of patients with chronic B-cell lymphoproliferative diseases. AB - BACKGROUND: The success of treatment using monoclonal antibodies in oncology is influenced by, among other factors, the level of target antigen expression on tumor cells. The authors analyzed the intensity of the CD52 antigen expression in patients with chronic lymphoproliferative diseases and compared them with B lymphocytes of a healthy population and CD34(+) cells in peripheral blood stem cells (PBSC) grafts. METHODS: Recently diagnosed and previously untreated patients with B-cell chronic lymphocytic leukemia (B-CLL), mantle-cell lymphoma (MCL), or small lymphocytic lymphoma (SLL) were evaluated and compared with control group and CD34(+) cells. The intensity of CD52 was expressed in molecules of equivalent soluble fluorochrome units (MESF) and antibody-binding capacity (ABC). RESULTS: In the group of patients with B-CLL, the CD52 level on tumor cells (245 x 10(3) MESF; 107 x 10(3) ABC) was significantly lower than on B lymphocytes of the control group (446 x 10(3) MESF; 194 x 10(3) ABC; P < 0.001) and SLL tumor cells (526 x 10(3) MESF; 229 x 10(3) ABC; P < 0.001). The CD52 antigen was expressed on a majority of CD34(+) cells, but its expression intensity was low (101 x 10(3) MESF; 44 x 10(3) ABC). CONCLUSIONS: Our data demonstrate differences in the intensity of the CD52 antigen expression between B lymphocytes and tumor lymphocytes of B-CLL patients, and between B-CLL and SLL tumor cells. CD52 antigen is expressed at low level on CD34(+) cells. PMID- 17428003 TI - Scaleable purification process for gene therapy retroviral vectors. AB - BACKGROUND: Retroviral vectors (RVs) constitute one of the preferred gene therapy tools against inherited and acquired diseases. Development of scaleable downstream processes allowing purification under mild conditions and yielding viral preparations with high titer, potency and purity is critical for the success of clinical trials and subsequent clinical use of this technology. METHODS: A purification process for murine leukaemia virus (MLV)-derived vector supernatants was developed based on membrane separation and anion-exchange chromatography (AEXc). Initial clarification of the vector stocks was performed using 0.45 microm membranes followed by concentration with 500 kDa molecular weight cut-off (MWCO) membranes; further purification was performed by AEXc using a tentacle matrix bearing DEAE functional ligands. Finally, concentration/diafiltration was performed by 500 kDa MWCO membranes. To validate final product quality the process was scaled up 16-fold. RESULTS: Optimization of microfiltration membrane pore size and ultrafiltration transmembrane pressure allowed the recovery of nearly 100% infectious particles. Further purification of the RVs by AEXc resulted in high removal of protein contaminants while maintaining high recoveries of infectious vectors (77+/-11%). Up-scaling of the process resulted in high titer vector preparations, 3.2x10(8) infectious particles (IP)/ml (85-fold concentration), with an overall recovery reaching 26%. The process yielded vectors with transduction efficiencies higher than the starting material and more than 99% pure, relative to protein contamination. CONCLUSIONS: The combination of membrane separation and AEXc processes results in a feasible and scaleable purification strategy for MLV-derived vectors, allowing the removal of inhibitory contaminants thus yielding pure vectors with increased transduction efficiencies. PMID- 17428004 TI - Analysis of chemical and metabolic components in traditional Chinese medicinal combined prescription containing Radix Salvia miltiorrhiza and Radix Panax notoginseng by LC-ESI-MS methods. AB - High-performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) methods were developed for the analysis of chemical and metabolic components in traditional Chinese medicinal combined prescription containing Radix Salvia miltiorrhiza and Radix Panax notoginseng (commonly known as Fufang Danshen prescription, FDP). The HPLC experiments used a reversed-phase Zorbax C(18) column with the column temperature at 30 degrees C and a binary mobile phase system consisting of aqueous formic acid (0.1%, v/v) and acetonitrile using a gradient elution at the flow rate of 1.0 mL/min. The ESI-MS was operated with a single-quadrupole mass spectrometer in both negative and positive ion modes. 36 major chromatographic peaks of FDP, including 14 saponins, 13 phenolic acids and nine diterpenoid quinones were characterized by their MS spectra and in comparison with some of the reference standards. In addition, after oral administration of extraction of FDP, the rat's plasma, urine and feces were also analyzed; 53 metabolic components including 30 original components and 23 transformative components of FDP were detected, and possible metabolic pathways of some components in FDP were given. The analysis of chemical and metabolic components in FDP by HPLC-MS methods could be a useful means of identifying the multi-components of FDP and to hint at their possible metabolic mechanism of action in the body. PMID- 17428005 TI - Mechanism of oleic acid-induced myofibril disassembly in rat cardiomyocytes. AB - This study investigated the mechanism of oleic acid (OA)-induced disassembly of myofibrils in cardiomyocytes. OA treatment disrupted myofibrils, as revealed by the disorganization of several sarcomeric proteins. Since focal adhesions (FAs) are implicated in myofibril assembly, we examined structural changes in FAs after OA treatment. Immunofluorescence studies with antibodies against FA proteins (vinculin, integrin beta1D, and paxillin) showed that FAs and costameres disintegrated or disappeared after OA treatment and that the changes in FA proteins occurred prior to myofibril disassembly. The effects of OA on FAs and myofibrils were reversed after removal of OA. OA decreased expression of integrin beta1D, paxillin, vinculin, and actin, and induced tyrosine dephosphorylation of FA kinase (FAK) and paxillin. These effects were blocked by pretreatment with sodium orthovanadate, a protein tyrosine phosphatase (PTP) inhibitor. This inhibitor also prevented OA-induced myofibril disassembly, indicating the involvement of PTP in myofibril disassembly. Furthermore, OA increased protein levels of PTP-PEST. The upregulation of this phosphatase correlated with the tyrosine dephosphorylation of paxillin and FAK, which are targets for PTP-PEST. In addition, OA decreased RhoA activity and the phosphorylation of cofilin, a downstream target of RhoA. Cofilin dephosphorylation increased its actin-severing activity and led to the depolymerization of F-actin, which might provide another potential mechanism for OA-induced myofibril disassembly. PMID- 17428007 TI - Post-harvest HPLC determination of chlorfluazuron residues in pears treated with different programs. AB - The present study was conducted to monitor the level of chlorfluazuron residues in pear samples in order to assess the risk posed by the presence of such residues to the consumer. Chlorfluazuron was sprayed onto pear trees at the recommended dose rate at two different times at 30 and 21 days prior to harvesting in one treatment, at 21 and 14 days prior to harvesting in another treatment, and three times at 30, 21 and 14 days prior to harvesting in a third treatment. Chlorfluazuron residues were extracted with acetonitrile and partitioned into ethyl acetate. The residue determination was performed on an Apollo C(18) column using HPLC with a UV detection of 254 nm following the clean up of the extract by open column chromatography with Florisil. The versatility of this method was evidenced by its good linearity (>0.995) in the concentration range between 0.2 and 10 microg/mL. The majority of the mean recoveries at two different fortification levels, 0.05 and 0.25 ppm, ranged from 84.9 +/- 3.2 to 94.3 +/- 10.6, and the repeatability (as the relative standard deviation) from three repetitive determinations of recovery was between 3.8 and 11%. The calculated limit of detection (LOD) was 0.008 ppm and the limit of quantitation was 0.03 ppm. Trace amounts of chlorfluazuron were detectable when it was applied onto the pear trees at two or three times prior to harvesting; however, the levels of chlorfluazuron were not quantified. The excellent sensitivity and selectivity of this method allowed for quantitation and identification at low levels with a run time of less than 12 min. Chlorfluazuron can be used safely to protect pears when sprayed two or three times at 14 days prior to harvesting. PMID- 17428006 TI - The relative importance of specific risk factors for insomnia in women treated for early-stage breast cancer. AB - BACKGROUND: Many individual risk factors for insomnia have been identified for women with a history of breast cancer. We assessed the relative importance of a wide range of risk factors for insomnia in this population. METHODS: Two thousand six hundred and forty-five women < or =4 years post-treatment for Stage I (> or =1 cm)-IIIA breast cancer provided data on cancer-related variables, personal characteristics, health behaviors, physical health/symptoms, psychosocial variables, and the Women's Health Initiative-Insomnia Rating Scale (WHI-IRS; scores > or =9 indicate clinically significant insomnia). RESULTS: Thirty-nine per cent had elevated WHI-IRS scores. In binary logistic regression, the variance in high/low insomnia group status accounted for by each risk factor category was: cancer-specific variables, 0.4% (n.s.); personal characteristics, 0.9% (n.s.); health behaviors, 0.6% (n.s.); physical health/symptoms, 13.4% (p<0.001); and, psychosocial factors, 11.4% (p<0.001). Insomnia was associated with worse depressive (OR = 1.32) and vasomotor symptoms (particularly night sweats) (OR = 1.57). CONCLUSION: Various cancer-specific, demographic, health behavior, physical health, and psychosocial factors have been previously reported as risk factors for insomnia in breast cancer. In our study (which was powered for simultaneous examination of a variety of variables), cancer-specific, health behavior, and other patient variables were not significant risk factors when in the presence of physical health and psychosocial variables. Only worse depressive and vasomotor symptoms were meaningful predictors. PMID- 17428008 TI - Determination of a selective Na+/H+ exchanger inhibitor, 4 cyano(benzo[b]thiophene-2-carbonyl)guanidine (KR-33028) in rat plasma by liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography-tandem mass spectrometric (LC/MS/MS) method was developed for the determination of a selective Na(+)/H(+) exchanger inhibitor 4 cyano(benzo[b]thiophene-2-carbonyl)guanidine (KR-33028) in rat plasma. KR-33028 and the internal standard, linezolid, were extracted from rat plasma with ethyl acetate at neutral pH. The analytes were separated on an XBridge C(18) column with a mixture of methanol-0.1% formic acid (35:65, v/v) as mobile phase and detected using an electrospray ionization tandem mass spectrometry in the multiple-reaction-monitoring mode. The standard curve was linear (r = 0.9998) over the concentration range of 2.0-1000 ng/mL. The coefficients of variation of intra- and inter-assay were 1.3-6.8% and the relative error was 0.8-5.0%. The recoveries of KR-33028 and linezolid were 70.5 and 84.6%, respectively. The lower limit of quantification for KR-33028 was 2.0 ng/mL using 50 microL plasma sample. This method was successfully applied to the pharmacokinetic study of KR-33028 in rats. PMID- 17428009 TI - Simultaneous quantitative and qualitative analysis of bioactive phenols in Dendrobium aurantiacum var. denneanum by high-performance liquid chromatography coupled with mass spectrometry and diode array detection. AB - A novel high-performance liquid chromatographic method with mass spectrometry and diode array detection method for the simultaneous qualitative and quantitative analysis of bioactive phenols was developed. In total, nine chemically diverse phenols including five bibenzyls, three phenanthrenes and a coumarin were unambiguously identified in Dendrobium aurantiacum var. denneanum by comparison with the available references or reported data according to their retention behaviors, UV spectra and fragmentations of ESI-MS. The contents of the four main phenolic compounds, moscatilin, gigantol, moscatin and coumarin, in D. aurantiacum var. denneanum from the wild and various cultivated populations were determined by HPLC-UV. The sample preparation involved a rapid and simple procedure based on solid-phase extraction using a C(18) reversed-phase cartridge. The quantitative analysis was performed on a Beckman Coulter ODS column (5 microm, 250 x 4.6 mm) using a linear gradient elution system of acetonitrile-0.5% formic acid. The method was validated for linearity, limits of detection (LOD) and quantification (LOQ), precision and accuracy. Good results were obtained with respect to the overall intra- and inter-day variations (RSD less than 3.22%) and the percentage recoveries (ranging from 90.50 to 99.22%). Notable differences in the contents of phenols were observed among different cultivated populations. The samples colleted in April and May (spring), or October and November (autumn) accumulated much higher contents of phenols than those collected in other seasons. PMID- 17428010 TI - Identification of the major metabolites of 5-n-butyl-4-{4-[2-(1H-tetrazole-5-yl) 1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2-(2,6-dichloridephenyl)-3H-1,2,4 triazol-3-one, a new angiotensin type 1 receptor antagonist, in rat bile by HPLC diode array detection-MS and HPLC-MS/MS. AB - 5-n-Butyl-4-{4-[2-(1H-tetrazole-5-yl)-1H-pyrrol-1-yl]phenylmethyl}-2,4-dihydro-2 (2,6-dichloridephenyl)-3H-1,2,4-triazol-3-one (1b), a new non-peptide angiotensin type 1 receptor antagonist, has been observed to play a positive role in the treatment of hypertension in preclinical tests. Rats were dosed with the drug, and the major metabolites in the bile were separated by gradient elution high performance liquid chromatography. HPLC-diode array detection-mass spectrometry, HPLC-electrospray ionization MS/MS methods in negative ion mode and collision induced dissociation were used to elucidate the structures of the major metabolites of 1b. One dihydroxylated 1b (M1), two monohydoxylated 1b (M2, M3) and one 1b monoglucuronide (M5) were found in male rat bile; however, three monohydoxylated 1b (M2, M3, M4) and one 1b monoglucuronide (M5) were found in female rat bile. These metabolites greatly differ in amount between male and female rat bile. PMID- 17428011 TI - Simultaneous quantification of oxysophocarpine and its active metabolite sophocarpine in rat plasma by liquid chromatography/mass spectrometry for a pharmacokinetic study. AB - A sensitive, rapid and specific method for the simultaneous quantification of oxysophocarpine (OSC) and its active metabolite sophocarpine (SC) in rat plasma was developed and validated, using a liquid-liquid extraction procedure followed by liquid chromatography/electrospray ionization mass spectrometric (LC/ESI-MS) analysis. The separation was performed on a Zorbax Extend-C(18) column (2.1 mm i.d. x 50 mm, 5 microm) with a C(18) guard column using methanol-water containing 5 mm ammonium acetate (15:85, v/v) as mobile phase. Analysis was performed in selected ion monitoring (SIM) mode with an electrospray ionization (ESI) interface. [M + H](+) at m/z 263 for OSC, [M + H](+) at m/z 247 for SC and [M + H](+) at m/z 249 for matrine (internal standard) were selected as detecting ions, respectively. The method was linear in the concentration ranges 10-1000 ng/mL for OSC and 5-500 ng/mL for SC. The intra- and inter-day precisions (coefficient of variation) were within 7% for both analytes. Their accuracy (relative error) ranged from -6.4 to 1.5%. The limits of detection for OSC and SC were 3 and 1.5 ng/mL, respectively. The limits of quantitation for OSC and SC were 10 and 5 ng/mL, respectively. Recoveries of both analytes were greater than 85% at the low, medium and high concentrations. Both analytes were stable during all sample storage, preparation and analytic procedures. The method was successfully applied to a pharmacokinetic study after an oral administration of OSC to rats with a dose of 15 mg/kg. PMID- 17428012 TI - Simultaneous qualitation and quantification of four phytoecdysones in Radix Achyranthis Bidentatae by high-performance liquid chromatography with diode array detection. AB - A high-performance liquid chromatographic method with diode array detection was developed and validated to simultaneously identify and quantify four phytoecdysones,i.e., polypodine B (1), ecdysterone (2), 25-R inokosterone (3) and 25-S inokosterone (4), in Radix Achyranthis Bidentatae. The analysis was performed using a C(18) column with 6% tetrahydrofuran aqueous and acetonitrile isocratic elution, and the detection was carried out by DAD at 242 nm. The identities of the analytes were determined by comparing the retention time and UV spectrum with those of reference compounds. The validation of the method included linearity, sensitivity, recovery and stability. All calibration curves of the four phytoecdysones showed good linear regression (r(2) >or= 0.9993). The limit of detection (S/N = 3) and limit of quantification (S/N = 10) were less than 7.5 and 12.3 ng, respectively. The method provided desirable repeatability with overall intra- and inter-day variations of less than 4.67%. The obtained recoveries varied between 95.1 and 104.4% while the relative standard deviations were below 4.85% (n = 3). The analysis results indicated that the contents of the investigated phytoecdysones in Radix Achyranthis Bidentatae from different locations were highly variant, and the genuine crude drug indigenous to Henan province did not possess the highest concentration of phytoecdysones. PMID- 17428013 TI - Performance evaluation of elemental analysis/isotope ratio mass spectrometry methods for the determination of the D/H ratio in tetramethylurea and other compounds--results of a laboratory inter-comparison. AB - Tetramethylurea (TMU) with a certified D/H ratio is the internal standard for Site-specific Natural Isotope Fractionation measured by Nuclear Magnetic Resonance (SNIF-NMR) analysis of wine ethanol for detection of possible adulterations (Commission Regulation 2676/90). A new batch of a TMU certified reference material (CRM) is currently being prepared. Whereas SNIF-NMR has been employed up to now, Elemental Analysis/Isotope Ratio Mass Spectrometry ((2)H-EA IRMS) was envisaged as the method of choice for value assignment of the new CRM, as more precise (better repeatable) data might be obtained, resulting in lower uncertainty of the certified value. In order to evaluate the accuracy and intra- and inter-laboratory reproducibility of (2)H-EA-IRMS methods, a laboratory inter comparison was carried out by analysing TMU and other organic compounds, as well as some waters. The results revealed that experienced laboratories are capable of generating robust and well comparable data, which highlights the emerging potential of IRMS in food authenticity testing. However, a systematic bias between IRMS and SNIF-NMR reference data was observed for TMU; this lack of data consistency rules out the (2)H-IRMS technique for the characterisation measurement of the new TMU CRM. PMID- 17428014 TI - Study on ion formation in electrospray droplet impact secondary ion mass spectrometry. AB - A new type of cluster secondary ion mass spectrometry (SIMS), named electrospray droplet impact (EDI), has been developed in our laboratory. In general, rather strong negative ions as well as positive ions can be generated by EDI compared with conventional SIMS. In this work, various aspects of ion formation in EDI are investigated. The Bronsted bases (proton acceptor) and acids (proton donor) mixed in the analyte samples enhanced the signal intensities of deprotonated molecules (negative ions) and protonated molecules (positive ions), respectively, for analytes. This suggests the occurrence of heterogeneous proton transfer reactions (i.e. M + M' --> [M+H](+) + [M'-H](-)) in the shockwave-heated selvedge of the colliding interface between the water droplet and the solid sample deposited on the metal substrate. EDI-SIMS shows a remarkable tolerance to the large excess of salts present in samples. The mechanism for desorption/ionization in EDI is much simpler than those for MALDI and SIMS because only very thin sample layers take part in the shockwave-heated selvedge and complicated higher-order reactions are largely suppressed. PMID- 17428015 TI - Fast and direct screening of polyaromatic hydrocarbon (PAH)-contaminated sand using a miniaturized membrane inlet mass spectrometer (mini-MIMS). AB - A miniaturized membrane inlet mass spectrometer (mini-MIMS; total weight 10 kg everything included) was equipped with a small sample cell using a flat sheet silicone membrane mounted close to the ionizing region of a multipole mass spectrometer. Spiked sand samples were placed in small stainless steel vials and dropped into the heated sample cell (>150 degrees C). A hole in the vial in front of the membrane and above the sand made it possible for the polyaromatic hydrocarbon (PAH) residuals to penetrate the membrane and enter the mass spectrometer as they evaporated from the sample. Using this simple setup we were able to quantitatively (approximately 10% relative standard deviation (RSD)) detect PAHs with up to five aromatic rings and with detection limits in the low parts-per-million (ppm) range. The vial system solves one of the major difficulties in analysis of larger PAHs using a MIMS. Normally, analysis of PAHs with more than two rings is hampered by a long memory effect due to the sticking of the PAHs to the inlet system, the membrane and surfaces in the vacuum system. By removing the vial from the sample cell within 2 min, we were able to analyze samples at 5-10 min intervals. The preliminary laboratory experiments presented here show much promise with respect to the development of a hand held (<10 kg) on site mass spectrometry system for PAH screening at contaminated sites. PMID- 17428016 TI - Electrospray ionization/atmospheric pressure photoionization multimode source for low-flow liquid chromatography/mass spectrometric analysis. AB - Analysis of several polar and non-polar compounds is performed with a newly developed dual electrospray ionization/atmospheric pressure photoionization (ESI/APPI) or ESPI source. Several variables are considered in the source, such as ESI probe heater temperature, solvent flow, dopant effects, repeller plate voltage, source geometry and photon energy (Kr vs. Ar lamp). Direct photoionization resulting in a molecular radical cation [M](*+) dominates at high temperatures (>400 degrees C) and low flow rates (<200 microL/min). Indirect photo-induced chemical ionization (PCI) involving solvent molecules becomes important at lower temperatures and higher solvent flow rates. Indirect PCI is enhanced using an Ar lamp, which yields comparable [M+H](+) signal but poorer [M](*+) signal than the Kr lamp at lower temperatures and higher flow rates. This is in support of our recent finding that the Ar lamp results in a solvent dependent enhancement of analyte molecules via PCI. Analysis of 12 compounds in methanol under low-flow conditions (10 microL/min) demonstrates that the dual ESPI source performs favorably for most compounds versus the standard ESCI source, and significantly better than ESCI for the analysis of unstable drugs, like flurbiprofen. Several factors contributing to the benefits of the ESPI source are the shared optimal geometry for ESI and APPI sources and soft ionization of APPI versus APCI. PMID- 17428017 TI - Validated LC-MS/MS method for quantification of gabapentin in human plasma: application to pharmacokinetic and bioequivalence studies in Korean volunteers. AB - A sensitive validated liquid chromatography-tandem mass spectrometric method (LC MS/MS) for gabapentin (GB) in human plasma has been developed and applied to pharmacokinetic (PK) and bioequivalence (BE) studies in human. In a randomized crossover design with a 1-week period, each subject received a 300 mg GB capsule. The procedure involves a simple protein precipitation with acetonitrile and separated by LC with a Gemini C(18) column using acetonitrile-10 mm ammonium acetate (20:80, v/v, pH 3.2) as mobile phase. The GB and internal standard [(S) (+)-alpha-aminocyclohexanepropionic acid hydrate] were analyzed using an LC-API 2000 MS/MS in multiple reaction monitoring mode. The ionization was optimized using ESI(+) and selectivity was achieved using MS/MS analysis, m/z 172.0 --> 154.0 and m/z 172.0 --> 126.0 for GB and IS, respectively. The assay exhibited good linearity over a working range of 20-5000 ng/mL for GB in human plasma with a lower limit of quantitation of 20 ng/mL. No endogenous compounds were found to interfere with the analysis. The accuracy and precision were shown for concentrations over the standard ranges. This method was successfully applied for the PK and BE studies by analysis of blood samples taken up to 36 h after an oral dose of 300 mg of GB in 24 healthy volunteers. PMID- 17428018 TI - Development and evaluation of a rapid lateral flow immunochromatographic strip assay for screening 19-nortestosterone. AB - The lateral flow strip test for 19-nortestosterone is one kind of immunochromatographic assay. Nitrocellulose membrane was separately immobilized with goat anti-rabbit IgG (control line) and 19-NT-OVA conjugate (test line). Anti-19-NT polyclonal antibody labeled with colloidal gold particles acted as the detector reagent. The assay is qualitatively, not quantitatively, judged with positive or negative result. We tested the sensitivity of the strip using spiked swine urine, and each specimen was independently measured by LC/MS/MS. The sensitivity, measure by eye, was determined to be 200 ng/mL. The assay time was less than 15 min, and so suitable for on-site rapid test. PMID- 17428019 TI - Liquid chromatography-tandem mass spectrometric method for determination of the anti-inflammatory compound vicenin-2 in the leaves of L. ericoides Mart. AB - This paper reports a rapid and sensitive method for determination of the anti inflammatory compound vicenin-2 in L. ericoides Mart. using liquid chromatography tandem mass spectrometry. Separation of the compound of interest was performed on a VP-ODS(18) (150 x 2 mm, Shimadzu, Japan) column and a pre-column packed with GPV-ODS C(18) (5 x 2 mm, Sigma-Aldrich, USA) with acetonitrile-water (15:85) mobile phase containing 2% acetic acid using isocratic flow at 0.5 mL/min for 2 min. Multiple-reaction monitoring of vicenin-2 was performed using electrospray positive ionization. The linear calibration curves were generated using a concentration range of 5-2500 ng/mL with correlation coefficients >0.99. The values of limit of detection and limit of quantitation were found to be 1 and 5 ng/mL, respectively. The method developed based on LC-ESIMS/MS is advantageous because it permits the rapid and selective detection of vicenin-2. Furthermore, the method can be easily applied to the routine analysis of vicenin-2 in plant extracts using a minimal amount of sample. PMID- 17428020 TI - Determination and pharmacokinetics of manidipine in human plasma by HPLC/ESIMS. AB - A sensitive HPLC/ESIMS method was established for the determination of manidipine in human plasma and pharmacokinetics study. After basified plasma with ammonia, manidipine and the internal standard (IS) (felodipine) were extracted with n hexane and separated on a Hypersil ODS2 column with a mobile phase of methanol-5 mm ammonium acetate solution containing 0.1% acetic acid (85:15, v/v). MS determination was performed by electrospray ionization in the selected ion monitoring mode. Manidipine was monitored at m/z 611.4 and IS at m/z 384. The assay had a calibration range from 0.2 to 20 ng/mL and a lower limit of quantification of 0.1 ng/mL. The method has been successfully applied to the pharmacokinetic study in healthy volunteers. PMID- 17428022 TI - Charge carrier transporting molecular materials and their applications in devices. PMID- 17428023 TI - Organic semiconducting oligomers for use in thin film transistors. PMID- 17428024 TI - Micro- and nanopatterning techniques for organic electronic and optoelectronic systems. PMID- 17428025 TI - Electronic and vibrational spectroscopies applied to organic/inorganic interfaces. PMID- 17428026 TI - Conjugated polymer-based organic solar cells. PMID- 17428027 TI - Turnip yellow mosaic virus as a chemoaddressable bionanoparticle. AB - Viruses and virus-like particles (VLPs) have been demonstrated to be robust scaffolds for the construction of nanomaterials. In order to develop new nanoprobes for time-resolved fluoroimmuno assays as well as to investigate the two-dimensional self-assembly of viruses and VLPs, the icosahedral turnip yellow mosaic virus (TYMV) was investigated as a potential building block in our study. TYMV is an icosahedral plant virus with an average diameter of 28 nm that can be isolated inexpensively in gram quantities from turnips or Chinese cabbage. There are 180 coat protein subunits per TYMV capsid. The conventional N hydroxysuccinimide-mediated amidation reaction was employed for the chemical modification of the viral capsid. Tryptic digestion with sequential MALDI-TOF MS analysis identified that the amino groups of K32 of the flexible N-terminus made the major contribution for the reactivity of TYMV toward N-hydroxysuccinimide ester (NHS) reagents. The reactivity was also monitored with UV-vis absorbance and fluorescence, which revealed that approximately 60 lysines per particle could be addressed. We hypothesized that the flexible A chain contains the reactive lysine because the crystal structure of TYMV has shown that chain A is much more flexible compared to B and C, especially at the N-terminal region where the Lys 32 located. In addition, about 90 to 120 carboxyl groups, located in the most exposed sequence, could be modified with amines catalyzed with 1-(3 dimethylaminopropyl-3-ethylcarbodiimide) hydrochloride (EDC) and sulfo-NHS. TYMV was stable to a wide range of reaction conditions and maintained its integrity after the chemical conjugations. Therefore, it can potentially be employed as a reactive scaffold for the display of a variety of materials for applications in many areas of nanoscience. PMID- 17428028 TI - Evaluation of a published in silico model and construction of a novel Bayesian model for predicting phospholipidosis inducing potential. AB - The identification of phospholipidosis (PPL) during preclinical testing in animals is a recognized problem in the pharmaceutical industry. Depending on the intended indication and dosing regimen, PPL can delay or stop development of a compound in the drug discovery process. Therefore, for programs and projects where a PPL finding would have adverse impact on the success of the project, it would be desirable to be able to rapidly identify and screen out those compounds with the potential to induce PPL as early as possible. Currently, electron microscopy is the gold standard method for identifying phospholipidosis, but it is low-throughput and resource-demanding. Therefore, a low-cost, high-throughput screening strategy is required to overcome these limitations and be applicable in the drug discovery cycle. A recent publication by Ploemen et al. (Exp. Toxicol. Pathol. 2004, 55, 347-55) describes a method using the computed physicochemical properties pKa and ClogP as part of a simple calculation to determine a compound's potential to induce PPL. We have evaluated this method using a set of 201 compounds, both public and proprietary, with known in vivo PPL-inducing ability and have found the overall concordance to be 75%. We have proposed simple modifications to the model rules, which improve the model's concordance to 80%. Finally, we describe the development of a Bayesian model using the same compound set and found its overall concordance to be 83%. PMID- 17428029 TI - Basis set exchange: a community database for computational sciences. AB - Basis sets are some of the most important input data for computational models in the chemistry, materials, biology, and other science domains that utilize computational quantum mechanics methods. Providing a shared, Web-accessible environment where researchers can not only download basis sets in their required format but browse the data, contribute new basis sets, and ultimately curate and manage the data as a community will facilitate growth of this resource and encourage sharing both data and knowledge. We describe the Basis Set Exchange (BSE), a Web portal that provides advanced browsing and download capabilities, facilities for contributing basis set data, and an environment that incorporates tools to foster development and interaction of communities. The BSE leverages and enables continued development of the basis set library originally assembled at the Environmental Molecular Sciences Laboratory. PMID- 17428030 TI - Polyelectrolyte complexes stabilize and controllably release vascular endothelial growth factor. AB - Angiogenesis has long been a desired therapeutic approach to improve clinical outcomes of conditions typified by ischemia. Vascular endothelial growth factor (VEGF) has demonstrated the ability to generate new blood vessels in vivo, but trials using intravenous delivery have not yet produced clinical success. Localized, sustained delivery of VEGF has been proven necessary to generate blood vessels as demonstrated by implantable, controlled release devices. Ultimately, nanoparticles delivered by intravenous injection may be designed to accumulate in target tissues and sustain the local VEGF concentration; however, injectable nanosuspensions that control the release of stabilized VEGF must first be developed. In this study, we utilize the heparin binding domain of VEGF to bind the polyanion dextran sulfate, resulting in an enhanced thermal stability of VEGF. Coacervation of the VEGF-bound dextran sulfate with selected polycations (chitosan, polyethylenimine, or poly-L-lysine) produced nanoparticles approximately 250 nm in diameter with high VEGF encapsulation efficiency (50 85%). Release of VEGF from these formulations persisted for >10 days and maintained high VEGF activity as determined by ELISA and a mitogenic bioassay. Chitosan-dextran sulfate complexes were preferred because of their biodegradability, desirable particle size ( approximately 250 nm), entrapment efficiency ( approximately 85%), controlled release (near linear for 10 days), and mitogenic activity. PMID- 17428031 TI - Mass spectrometry imaging of small molecules using desorption/ionization on silicon. AB - Development of novel tools to image spatial distribution of small molecules in biological samples is essential in disease diagnosis and biomarker discovery. To simplify sample preparation and reduce background noise in the low-mass region, we describe here the use of a matrix-free mass spectrometric imaging method, i.e., desorption/ionization on silicon (DIOS), for biological surface analysis. The imaging parameters, such as the laser beam diameter and the translation stage movement, were studied and optimized to improve imaging performance. The use of DIOS imaging to map small molecules on mouse liver tissues was demonstrated. In addition, phosphatidylcholine (PC) and propidium iodide (PI) were used as the cell membrane and nucleus markers, respectively, to "visualize" the presence of HEK 293 cells. The reconstructed ion maps of PC and PI were compared with the optical images collected from the same sample using bright-field and fluorescence microscopy. A good correlation of the spatial distribution of cells confirmed the validity of this DIOS imaging approach. PMID- 17428032 TI - Secondary ion MS imaging to relatively quantify cholesterol in the membranes of individual cells from differentially treated populations. AB - Time-of-flight secondary ion mass spectrometry (TOF-SIMS) is a well-established bioanalytical method for directly imaging the chemical distribution across single cells. Here we report a protocol for the use of SIMS imaging to comparatively quantify the relative difference in cholesterol level between the plasma membranes of two cells. It should be possible to apply this procedure to the study of other selected lipids. This development enables direct comparison of the chemical effects of different drug treatments and incubation conditions in the plasma membrane at the single-cell level. Relative, quantitative TOF-SIMS imaging has been used here to compare macrophage cells treated to contain elevated levels of cholesterol with respect to control cells. In situ fluorescence microscopy with two different membrane dyes was used to discriminate morphologically similar but differentially treated cells prior to SIMS analysis. SIMS images of fluorescently identified cells reveal that the two populations of cells have distinct outer leaflet membrane compositions with the membranes of the cholesterol-treated macrophages containing more than twice the amount of cholesterol of control macrophages. Relative quantification with SIMS to compare the chemical composition of single cells can provide valuable information about normal biological functions, causative agents of diseases, and possible therapies for diseases. PMID- 17428033 TI - Electroosmosis-based nanopipettor. AB - Decreasing the volume of reagent solutions consumed in each assay is an effective means to reduce the overall cost in high-throughput analysis laboratories. Recently, increasing attention has been paid to investigate the behavior of individual cells. If one wishes to transfer solution to or from a single cell, a picoliter pipettor is needed since the entire cell volume is commonly less than 1 nL. While pressure ejection and iontophoresis have been used to deliver picoliter volumes of solutions, these techniques cannot yield routine pipettors which perform both solution "picking up" and "dispensing" functions. The state-of-the art pipettors can handle liquids down to approximately 100 nL, although the pipetting accuracy and precision deteriorate considerably from microliters to nanoliters. If one wishes to pipet reagents of less than 100 nL, new pipettors need to be developed. Electroosmosis has been utilized to pump solutions at flow rates of nanoliters to approximately picoliters per second, which is ideal for nanopipettors. The issue is how to arrange fluidic/electrical connections so that pipetting functions can be performed conveniently. In this paper, we present the results of our initial attempt to develop an electroosmosis-based nanopipettor. The first version of this pipettor consists of a microfabricated electroosmotic (EO) flow pump, a polyacrylamide grounding interface, and a nanoliter-to picoliter pipet tip. The detailed configuration and fabrication process of the pipettor are discussed. An excellent feature of an EO-driven pipettor is that it has no moving parts. Good reproducibilities (RSD = 6% at 140 pL, 2% at 950 pL, and 2% at 13 nL) and accuracies (9% at 0.13 nL, 4% at 1.0 nL, and 3% at 10 nL) of this pipettor have been demonstrated to aliquot/transport nanoliter-to-picoliter solutions. PMID- 17428034 TI - Trace analysis of DNA: preconcentration, separation, and electrochemical detection in microchip electrophoresis using Au nanoparticles. AB - We have developed a simple and sensitive on-chip preconcentration, separation, and electrochemical detection (ED) method for trace analysis of DNA. The microchip comprised of three parallel channels: the first two are for the field amplified sample stacking and subsequent field-amplified sampled injection steps, while the third one is for the microchip gel electrophoresis (MGE) with ED (MGE ED). To improve preconcentration and separation performances of the method, the stacking and separation buffers containing the hydroxypropyl cellulose (HPC) matrix were modified with gold nanoparticles (AuNPs). The formation of AuNPs and HPC/AuNP-modified buffers were characterized by UV-visible spectroscopy and TEM experiments. The conducting polymer-modified electrode was also modified with AuNPs to enhance detection performances of the electrode. The conducting polymer/AuNP layers act as electrocatalysts for the direct detection of DNA based on their oxidation in a solution phase. The total sensitivity was improved by approximately 25 000-fold when compared with a conventional MGE-ED analysis. The calibration plots were linear (r2 = 0.9993) within the range of 0.003-1.0 pg/microL for a 20-bp DNA sample. The sensitivity was 0.20 nA/(fg/microL), with a detection limit of 5.7 amol in a 50-microL sample, based on S/N = 3. The applicability of the method for the analysis of 13 fragments present in a 100-bp DNA ladder was successfully demonstrated. PMID- 17428035 TI - Kinetic and spectroscopic characterization of type II isopentenyl diphosphate isomerase from Thermus thermophilus: evidence for formation of substrate-induced flavin species. AB - Type II isopentenyl diphosphate (IPP) isomerase catalyzes the interconversion of IPP and dimethylallyl diphosphate (DMAPP). Although the reactions catalyzed by the type II enzyme and the well-studied type I IPP isomerase are identical, the type II protein requires reduced flavin for activity. The chemical mechanism, including the role of flavin, has not been established for type II IPP isomerase. Recombinant type II IPP isomerase from Thermus thermophilus HB27 was purified by Ni2+ affinity chromatography. The aerobically purified enzyme was inactive until the flavin cofactor was reduced by NADPH or dithionite or photochemically. The inactive oxidized flavin-enzyme complex bound IPP in a Mg2+-dependent manner for which KD approximately KmIPP, suggesting that the substrate binds to the inactive oxidized and active reduced forms of the protein with similar affinities. N,N Dimethyl-2-amino-1-ethyl diphosphate (NIPP), a transition state analogue for the type I isomerase, competitively inhibits the type II enzyme, but with a much lower affinity. pH-dependent spectral changes indicate that the binding of IPP, DMAPP, and a saturated analogue isopentyl diphosphate promotes protonation of anionic reduced flavin. Electron paramagnetic resonance (EPR) and UV-visible spectroscopy show a substrate-dependent accumulation of the neutral flavin semiquinone during both the flavoenzyme reduction and reoxidation processes in the presence of IPP and related analogues. Redox potentials of IPP-bound enzyme indicate that the neutral semiquinone state of the flavin is stabilized thermodynamically relative to free FMN in solution. PMID- 17428036 TI - FTIR study of the retinal Schiff base and internal water molecules of proteorhodopsin. AB - Proteorhodopsin (PR), an archaeal-type rhodopsin found in marine bacteria, is a light-driven proton pump similar to bacteriorhodopsin (BR). It is known that Asp97, a counterion of the protonated Schiff base, possesses a higher pKa ( approximately 7) compared to that of homologous Asp85 in BR (<3). This suggests that PR has a hydrogen-bonding network different from that of BR. We previously reported that a strongly hydrogen-bonded water molecule is observed only in the alkaline form of PR, where Asp97 is deprotonated (Furutani, Y., Ikeda, D., Shibata, M., and Kandori, H. (2006) Chem. Phys. 324, 705-708). This is probably correlated with the pH-dependent proton pumping activity of PR. In this work, we studied the water-containing hydrogen-bonding network in the Schiff base region of PR by means of Fourier-transform infrared (FTIR) spectroscopy at 77 K. [zeta 15N]Lys-labeling and 18O water were used for assigning the Schiff base N-D and water O-D stretching vibrations in D2O, respectively. The frequency upshift of the N-D stretch in the primary K intermediate is much smaller for PR than for BR, indicating that the Schiff base forms a hydrogen bond after retinal photoisomerization. We then measured FTIR spectra of the mutants of Asp97 (D97N and D97E) and Asp227 (D227N and D227E) to identify the amino acid interacting with the Schiff base in the K state. The PRK minus PR spectra of D97N and D97E were similar to those of the acidic and alkaline forms, respectively, of the wild type implying that the structural changes upon retinal photoisomerization are not influenced by the mutation at Asp97. In contrast, clear spectral differences were observed in D227N and D227E, including vibrational bands of the Schiff base and water molecules. It is concluded that Asp227 plays a crucial role during the photoisomerization process, though Asp97 acts as the primary counterion in the unphotolyzed state of PR. PMID- 17428037 TI - Ultrafast relaxation dynamics of perchlorinated cycloheptatriene in solution. AB - The photochemistry of perchlorinated cycloheptatriene (CHTCl(8)) has been studied by means of ultrafast pump-probe, transient anisotropy and continuous UV irradiation experiments in various solvents as well as by DFT calculations. After UV-excitation to the 1A' '-state, two competing reactions occur--a [1,7] sigmatropic chlorine migration via two ultrafast internal conversions and a [4,5] electrocyclization forming octachlorobicylo[3.2.0]hepta-[2,6]-diene. The first reaction has been studied by excitation with a 263 nm femtosecond-laser pulse. Pump-probe experiments reveal a first, solvent-independent time constant, tau1(CHTCl(8)) = 140 fs, that can be associated with the electronic relaxation of the 2A'-1A' ' transition, while a second one, tau2(CHTCl(8)), ranges from 0.9 to 1.8 ps depending on the polarity of the solvent. This finding is consistent with a [1,7]-chlorine migration during the 1A'-2A' transition where the migrating chlorine atom is partly negatively charged. The charge separation has also been confirmed by DFT calculations. Transient anisotropy measurements result in a time zero value of r(0) = 0.35 after deconvolution and a decay constant of tau1(a) = 120 fs, which can be explained by vibrational motions of CHTCl(8) in the electronically excited states, 1A' ' and 2A'. After continuous UV-irradiation of CHTCl(8), octachlorobicylo[3.2.0]hepta-[2,6]-diene is primarily formed with a solvent-dependent yield. From these investigations, we suggest a relaxation mechanism for CHTCl(8) after photoexcitation that is comparable to cycloheptatriene. PMID- 17428038 TI - Unraveling the formation of HCPH(X2A') molecules in extraterrestrial environments: crossed molecular beam study of the reaction of carbon atoms, C(3Pj), with phosphine, PH3(X1A1). AB - The reaction between ground state carbon atoms, C(3P(j)), and phosphine, PH3(X(1)A1), was investigated at two collision energies of 21.1 and 42.5 kJ mol( 1) using the crossed molecular beam technique. The chemical dynamics extracted from the time-of-flight spectra and laboratory angular distributions combined with ab initio calculations propose that the reaction proceeds on the triplet surface via an addition of atomic carbon to the phosphorus atom. This leads to a triplet CPH3 complex. A successive hydrogen shift forms an HCPH2 intermediate. The latter was found to decompose through atomic hydrogen emission leading to the cis/trans-HCPH(X(2)A') reaction products. The identification of cis/trans HCPH(X(2)A') molecules under single collision conditions presents a potential pathway to form the very first carbon-phosphorus bond in extraterrestrial environments like molecular clouds and circumstellar envelopes, and even in the postplume chemistry of the collision of comet Shoemaker-Levy 9 with Jupiter. PMID- 17428039 TI - Diketone radical cations: ketonic and enolic forms as revealed by matrix EPR studies and DFT calculations. AB - Radical cations of 2,3-butanedione, 2,4-pentanedione, 3-methylpentane-2,4-dione, 2,5-hexanedione, and 2,3-pentanedione were investigated by electron paramagnetic resonance (EPR) spectroscopy in a solid Freon matrix and density functional theory (DFT) quantum chemical calculations. All the diketone radical cations in ketonic form show small proton hyperfine couplings (typically unresolved in the EPR spectra). In the cases of 2,4-pentanedione and 3-methylpentane-2,4-dione, enolic forms of the radical cations (pi-type species with main spin population at carbon atom) were characterized. Preferential stabilization of the enolic form of 3-methylpentane-2,4-dione radical cation was explained by trap-to-trap positive hole migration rather than monomolecular relaxation of the ionized ketonic form through H atom transfer. PMID- 17428040 TI - Vibrational spectra, ab initio calculations, and ring-puckering potential energy function for gamma-crotonolactone. AB - The infrared and Raman spectra of liquid and vapor gamma-crotonolactone have been collected. Both the experimental data and ab initio calculations show that the molecule is rigidly planar in its electronic ground state. This conclusion agrees with the previously reported microwave studies and is attributed to the conjugation between the C=C and C=O double bonds of the ring. The ring-puckering potential energy function was generated from ab initio calculations and was confirmed by the vapor-phase Raman spectra to be nearly harmonic. Density functional theory (DFT) calculations predict a harmonic ring-puckering frequency of 203 cm(-1) as compared to the observed vapor-phase Raman value of 208 cm(-1). The DFT calculations were also used to compute the infrared and Raman spectra of gamma-crotonolactone, and these agree very well with the experimental spectra. PMID- 17428041 TI - Treatment of collinear and noncollinear electron spin within an approximate density functional based method. AB - We report benchmark calculations of the density functional based tight-binding method concerning the magnetic properties of small iron clusters (Fe2 to Fe5) and the Fe13 icosahedron. Energetics and stability with respect to changes of cluster geometry of collinear and noncollinear spin configurations are in good agreement with ab initio results. The inclusion of spin-orbit coupling has been tested for the iron dimer. PMID- 17428042 TI - Initial steps toward automating the fitting of DFTB Erep(r). AB - The most time-consuming part of developing new parametrizations for the density functional based tight-binding (DFTB) method consists of producing accurate and transferable repulsive pair potentials. In the conventional approach to repulsive parametrization, every possible diatomic combination of the elements covered by the parametrization must be individually hand-constructed. We present an initial attempt to automate some of this time-consuming process. We consider a simple genetic algorithm-based approach to the fitting problem. PMID- 17428043 TI - Dihydroxypyrimidine-4-carboxamides as novel potent and selective HIV integrase inhibitors. AB - Human immunodeficiency virus type-1 (HIV-1) integrase, one of the three constitutive viral enzymes required for replication, is a rational target for chemotherapeutic intervention in the treatment of AIDS that has also recently been confirmed in the clinical setting. We report here on the design and synthesis of N-benzyl-5,6-dihydroxypyrimidine-4-carboxamides as a class of agents which exhibits potent inhibition of the HIV-integrase-catalyzed strand transfer process. In the current study, structural modifications on these molecules were made in order to examine effects on HIV-integrase inhibitory potencies. One of the most interesting compounds for this series is 2-[1-(dimethylamino)-1 methylethyl]-N-(4-fluorobenzyl)-5,6-dihydroxypyrimidine-4-carboxamide 38, with a CIC95 of 78 nM in the cell-based assay in the presence of serum proteins. The compound has favorable pharmacokinetic properties in preclinical species (rats, dogs, and monkeys) and shows no liabilities in several counterscreening assays, highlighting its potential as a clinically useful antiviral agent. PMID- 17428044 TI - Structure and magnetic properties of Pb2Cu3B4O11: a new copper borate featuring [Cu3O8]10- units. AB - Pb2Cu3B4O11 crystallizes in the monoclinic space group P2/n (No. 13) with a = 6.8016(15) A, b = 4.7123(10) A, c = 14.614(3) A, beta = 97.089(3) degrees, and Z = 2. The crystal structure consists of infinite [Cu3O8]10- zigzag chains of alternating dimers and monomers. The magnetic susceptibility and specific heat capacity show spin-gap and Curie-Weiss behaviors that can be explained by a model of Cu(2)-Cu(2) dimers and isolated or weakly coupled Cu(1) monomers. PMID- 17428045 TI - Nature of "hydrogen bond" in the diborane-benzene complex: covalent, electrostatic, or dispersive? AB - Motivated by the recent discovery of unusual "hydrogen bonding"-like interaction between a borane system and benzene molecules in a molecular crystal, we carried out quantum mechanical calculations on a model complex, diborane-benzene cluster. The aim is to understand the nature of this unique interaction, which is expected to play an essential role in this novel class of molecular crystals. As analyzed in the present study, the interaction between diborane and benzene is special in the following aspects: (1) this interaction is mostly dispersive; (2) the observed pseudodirectionality with one of the diborane bridge hydrogen directed toward the benzene centroid minimizes the van der Waals contact; and (3) in the "hydrogen bond" map, this interaction is located in a unique region, which is presently populated by a few known molecular complexes with very different chemical characteristics. It is anticipated that the results from the present analysis will provide meaningful guidance for molecular engineering with diborane benzene as a building block and for stabilization of this and possible other hydrogen bonds by dispersive contributions. PMID- 17428046 TI - The importance of questioning scientific assumptions: some lessons from f element chemistry. AB - As scientists, we know that we should constantly question the assumptions upon which our research is based. We also know that we do not do this often enough. The recent results in f element chemistry described here should serve to remind us not to take the traditional boundaries of any area of chemistry for granted including topics as fundamental as redox chemistry and bond-length generalizations. New ways of doing reductive chemistry in the f element area as well as the synthesis of "long bond organometallics" that have unconventional bond distances and reactivity demonstrate how the "rules" in this area, thought to be true for decades, have been recently overturned. The synthetic chemistry that made these advances possible has generated additional unexpected opportunities in f element chemistry that are also described here. Overall, these results should stimulate researchers in all areas to challenge their assumptions. PMID- 17428047 TI - In search of cyclohexane-like Sn6(12-): synthesis of Li2Ln5Sn7 (Ln = Ce, Pr, Sm, Eu) with an open-chain heptane-like Sn7(16-) instead. AB - The title compounds were prepared by direct reactions of the corresponding elements at high temperature. They are isostructural and crystallize in the chiral orthorhombic space group P212121 (Li2Ce5Sn7: a = 6.273(1), b = 13.839(2), and c = 17.467(2) A; Li2Pr5Sn7: a = 6.241(1), b = 13.762(2), and c = 17.367(1) A; Li2Sm5Sn7: a = 6.262 (1), b = 13.809(1), and c = 17.432(1) A; Li2Eu5Sn7: a = 6.165(1), b = 13.562(2), and c = 17.128(1) A). The structure contains isolated Sn7 oligomers that resemble the carbon core of an open-chain heptane molecule C7H16. Although these heptamers are stacked along the a axis at a distance that is comparable to the distances within the heptamer, electronic structure calculations show that this intermolecular contact is nonbonding for a formal charge of 16- or higher per heptamer. A hypothetical lower charge of 14-, on the other hand, leads to positive and substantial bond-overlap population that would result in branched infinite chains of infinity[Sn714-]. Magnetic measurements of the Ce and Pr compounds indicate a 3+ oxidation state for the rare-earth cations and, therefore, 17 available electrons from the cations per formula unit. According to four-probe conductivity measurements, the compounds are metallic. PMID- 17428048 TI - Dimerization processes of square planar [PtII(tbpy)(dithiolato*)]+ radicals. AB - The preparation and structural characterization of the neutral, square planar complexes [PtII(tbpy)(A)] (1), [PtII(tbpy)(B)] (2), and [PtII(PPh3)2(B)] (3) have been accomplished, where (tbpy) = 4,4'-di-tert-butylpyridine, (A)2- = 3,6 bis(trimethylsilyl)-1,2-benzenedithiolate(2-), and (B)2- = 1,2-bis(4-tert butylphenyl)ethylene-1,2-dithiolate(2-) and (A*)1- and (B*)1- represent the corresponding monoanionic radicals. Electrochemical and chemical one-electron oxidation of 1 and 2 in CH2Cl2 solution affords the monomeric monocations [PtII(tbpy)(A*)]+ (1a) and [PtII(tbpy)(B*)]+ (2a), both of which possess an S = 1/2 ground state. The corresponding spin doublet monocationic dimers [PtII2(tbpy)2(A)(A*)]+ (1b) and [PtII2(tbpy)2(B)(B*)]+ (2b) were electrochemically generated in solution (50% oxidation) and identified by X-band EPR spectroscopy. Complete one-electron oxidation of 1 and 2 yielded the diamagnetic dimers [PtII2(tbpy)2(A*)2]2+ (1c) and [PtII2(tbpy)2(B*)2]2+ (2c) which are in equilibrium with the corresponding paramagnetic monomers 1a and 2a in solution. The crystal structure of [PtII2(tbpy)2(B*)2](PF6)2.3CH2Cl2 (2c) revealed a centrosymmetric, lateral dimer whose bridging part is a PtII2(mu2-S)2 rhomb; the metal ions possess a square based pyramidal geometry. Solid-state sulfur K-edge X-ray absorption spectra of 1, 2, 2a, 2c, and [PtII(B*)2]0 (4) have been recorded, which clearly show that a sulfur-centered radical (B*)1- is present in 2a, 2c, and 4. The absence of ligand-based radicals in 1 and 2 is also clearly established. One-electron oxidation of [Pt(PPh3)2(B)] (3) afforded only the spin doublet species [PtII(PPh3)2(B*)]+ (3a); no dimer formation was detected. Synthesis and crystal structure of square planar [PtII(B*)2]0.thf are also reported. PMID- 17428049 TI - Tunneling in green tea: understanding the antioxidant activity of catechol containing compounds. A variational transition-state theory study. AB - The catechol functionality present in the catechins is responsible for the protective effects exerted by green tea against a wide range of human diseases. High-level electronic structure calculations and canonical variational transition state theory including multidimensional tunneling corrections have allowed us to understand the key factors of the antioxidant effectiveness of the catechol group. This catechol group forms two hydrogen bonds with the two oxygen atoms of the lipid peroxyl radical, leading to a very compact reactant complex. This fact produces an extremely narrow adiabatic potential-energy profile corresponding to the hydrogen abstraction by the peroxyl radical, which makes it possible for a huge tunneling contribution to take place. So, quantum-mechanical tunneling highly increases the corresponding rate constant value, in such a way that catechins become able to trap the lipid peroxyl radicals in a dominant competition with the very damaging free-radical chain-lipid peroxidation reaction. PMID- 17428050 TI - Heparin/poly(l-lysine) nanoparticle-coated polymeric microspheres for stem-cell therapy. PMID- 17428051 TI - Concerted proton-electron transfer reactions in water. are the driving force and rate constant depending on pH when water acts as proton donor or acceptor? AB - The competition between stepwise and concerted (CPET) pathways in proton-coupled electron-transfer reactions in water is discussed on thermodynamic and kinetic bases. In the case where water is the proton acceptor, the CPET pathway may compete favorably with the stepwise pathway. The main parameter of the competition is pK of the oxidized form of the substrate being smaller or larger than 0. The driving force of the forward reaction is however independent of pH, despite the equilibrium redox potential of the proton-electron system being a function of pH. At high pH values, CPET reactions involving OH- as proton acceptor may likewise compete favorably with stepwise pathways. The overall reaction rate constant is an increasing function of pH, not because the driving force depends on pH but because OH- is a reactant. In buffered media, association of the substrate with the basic components of the buffer offers an alternative CPET route; the driving force comes closer to that offered by the pH-dependent equilibrium redox potential. PMID- 17428053 TI - Organoborane acceptor-substituted polythiophene via side-group borylation. PMID- 17428052 TI - Infrared spectroscopy and theoretical studies on gas-phase protonated leu enkephalin and its fragments: direct experimental evidence for the mobile proton. AB - The gas-phase structures of the protonated pentapeptide Leu-enkephalin and its main collision-induced dissociation (CID) product ions, b4 and a4, are investigated by means of infrared multiple-photon dissociation (IR-MPD) spectroscopy and detailed molecular mechanics and density functional theory (DFT) calculations. Our combined experimental and theoretical approach allows accurate structural probing of the site of protonation and the rearrangement reactions that have taken place in CID. It is shown that the singly protonated Leu enkephalin precursor is protonated on the N-terminus. The b4 fragment ion forms two types of structures: linear isomers with a C-terminal oxazolone ring, as well as cyclic peptide structures. For the former structure, two sites of proton attachment are observed, on the N-terminus and on the oxazolone ring nitrogen, as shown in a previous communication (Polfer, N. C.; Oomens, J.; Suhai, S.; Paizs, B. J. Am. Chem. Soc. 2005, 127, 17154-17155). Upon leaving the ions for longer radiative cooling delays in the ion cyclotron resonance (ICR) cell prior to IR spectroscopic investigation, one observes a gradual decrease in the relative population of oxazolone-protonated b4 and a corresponding increase in N-terminal protonated b4. This experimentally demonstrates that the mobile proton is transferred between two sites in a gas-phase peptide ion and allows one to rationalize how the proton moves around the molecule in the dissociation process. The a4 fragment, which is predominantly formed via b4, is also confirmed to adopt two types of structures: linear imine-type structures, and cyclic structures; the former isomers are exclusively protonated on the N-terminus in sharp contrast to b4, where a mixture of protonation sites was found. The presence of cyclic b4 and a4 fragment ions is the first direct experimental proof that fully cyclic structures are formed in CID. These results suggest that their presence is significant, thus lending strong support to the recently discovered peptide fragmentation pathways (Harrison, A. G.; Young, A. B.; Bleiholder, B.; Suhai, S.; Paizs, B. J. Am. Chem. Soc. 2006, 128, 10364-10365) that result in scrambling of the amino acid sequence upon CID. PMID- 17428054 TI - Ligand conjugated low-density lipoprotein nanoparticles for enhanced optical cancer imaging in vivo. PMID- 17428055 TI - Dielectric anisotropy of a homochiral trinuclear nickel(II) complex. PMID- 17428056 TI - Gold catalysts open a new general chemoselective route to synthesize oximes by hydrogenation of alpha,beta-unsaturated nitrocompounds with H2. PMID- 17428057 TI - Organic-inorganic hybrids based on pyrene functionalized octavinylsilsesquioxane cores for application in OLEDs. PMID- 17428058 TI - A vaulted biaryl phosphoric acid-catalyzed reduction of alpha-imino esters: the highly enantioselective preparation of alpha-amino esters. PMID- 17428059 TI - Highly enantioselective insertion of carbenoids into N-H bonds catalyzed by copper complexes of chiral spiro bisoxazolines. PMID- 17428060 TI - Enantioselective organocatalytic intramolecular ring-closing Friedel-Crafts-type alkylation of indoles. AB - An enantioselective organocatalytic intramolecular ring-closing Friedel-Crafts type alkylation of indolyl alpha,beta-unsaturated aldehydes has been developed. This powerful new strategy allows enantioselective access to THPIs and THBCs in a straightforward and atom-economical manner. PMID- 17428061 TI - Gold(I)-catalyzed intramolecular enantioselective hydroarylation of allenes with indoles. AB - Treatment of 2-allenyl indole 4 with a catalytic 1:2 mixture of [(S)-2]Au2Cl2 [(S)-2 = (S)-3,5-tBu-4-MeO-MeOBIPHEP] and AgBF4 in toluene at -10 degrees C for 17 h led to isolation of tetrahydrocarbazole 5 in 88% yield with 92% ee. The protocol was effective for the cyclization of terminally disubstituted allenes and for the formation of seven-membered rings. PMID- 17428062 TI - Efficient removal of ruthenium byproducts from olefin metathesis products by simple aqueous extraction. AB - Simple aqueous extraction removed ruthenium byproducts efficiently from ring closing metathesis (RCM) reactions catalyzed by a poly(ethylene glycol) (PEG) supported N-heterocyclic carbene-based ruthenium complex. PMID- 17428063 TI - Attractive noncovalent interactions in the mechanism of grubbs second-generation Ru catalysts for olefin metathesis. AB - Second-generation ruthenium carbenoid catalysts for olefin metathesis are a hundred to a thousand times more active than first-generation catalysts, despite a slower initiation step. A new density functional capable of treating medium range correlation energy shows that the relative rates of generation of the catalyst are determined by attractive noncovalent interactions. PMID- 17428064 TI - A new construction of 2-alkoxypyrans by an acylation-reductive cyclization sequence. AB - A new convergent synthetic approach to a pyran motif common to many naturally occurring structures is described. In this approach, two fragments are joined by esterification, and a subsequent intramolecular reductive cyclization affords the 2-hydroxypyran. PMID- 17428065 TI - Bioactive 2-oxazolines: a new approach via one-pot, four-component reaction. AB - Substituted 2-oxazolines of the general structure shown above are found in several families of bioactive natural products and can be prepared in an efficient and general one-pot, four-component condensation. PMID- 17428066 TI - Affinity purification of copper chelating peptides from chickpea protein hydrolysates. AB - Chickpea protein hydrolysates obtained with alcalase and flavourzyme were used for purification of copper chelating peptides by affinity chromatography using copper immobilized on solid supports. The chelating activity of purified peptides was indirectly measured by the inhibition of beta-carotene oxidation in the presence of copper. Two protein hydrolysates, obtained after 10 and 100 min of hydrolysis, were the most inhibitory of beta-carotene oxidation. Purified copper chelating peptides from these protein hydrolysates contained 19.7 and 35.1% histidine, respectively, in comparison to 2.7 and 2.6% in the protein hydrolysates. Chelating peptides from hydrolysate obtained after 10 min of hydrolysis were the most antioxidative being 8.3 times more antioxidative than the hydrolysate, while chelating peptides purified from protein hydrolysate obtained after 100 min were 3.1 times more antioxidative than its hydrolysate. However, the histidine content was higher in peptides derived from the 100 min hydrolysate (19.7 against 35.1% in 10 min hydrolysate), indicating that this amino acid is not the only factor involved in the antioxidative activity, and other factors such as peptide size or amino acid sequence are also determinant. This manuscript shows that affinity chromatography is a useful procedure for purification of copper chelating peptides. This method can be extended to other metals of interest in nutrition, such as calcium, iron, or zinc. Purified chelating peptides, in addition to their antioxidative properties, may also be useful in food mineral fortification for increasing the bioavailability of these metals. PMID- 17428067 TI - Temporal sequence of cell wall disassembly events in developing fruits. 1. Analysis of raspberry (Rubus idaeus). AB - Raspberry fruits were harvested at five developmental stages, from green to red ripe, and the changes in cell wall composition, pectin and hemicellulose solubilization, and depolymerization were analyzed. Fruit softening at intermediate stages of ripening was associated with increased pectin solubilization, which occurred without depolymerization. Arabinose was found to be the most abundant noncellulosic neutral sugar in the cell wall and showed dramatic solubilization late in ripening. No changes in pectin molecular size were observed even at the 100% red stage. Subsequently, as fruit became fully ripe a dramatic depolymerization occurred. In contrast, the hemicellulosic fractions showed no significant changes in content or polymer size during ripening. The paper discusses the sequence of events leading to cell wall disassembly in raspberry fruit. PMID- 17428068 TI - Temporal sequence of cell wall disassembly events in developing fruits. 2. Analysis of blueberry (Vaccinium species). AB - Softening and pathogen susceptibility are the major factors limiting the marketing of blueberries as fresh fruits, and these traits are associated with fruit cell wall structure. However, few studies that characterize wall modifications occurring during development and ripening have been reported for this fruit. In this study the ripening-associated modifications of blueberry fruit cell walls (composition, pectin and hemicellulose solubilization, and depolymerization) at five stages of ripeness have been analyzed. Xylose was found to be the most abundant noncellulosic neutral sugar associated with fruit walls, and the observed high Xyl/Glc ratio suggested that xylans, which are usually a minor hemicellulosic fruit wall component, are abundant in blueberry. The pectic matrix showed increased solubilization at early and intermediate stages of ripening, but no changes were detected in late ripening. Furthermore, little reduction in pectin polymer size occurred during blueberry ripening. In contrast, hemicellulose levels decreased as ripening progressed, and a clear depolymerization of these components was observed. A model for cell wall degradation in this fruit is discussed. PMID- 17428069 TI - Mutagenesis of the supF gene by stereoisomers of 1,2,3,4-diepoxybutane. AB - 1,2,3,4-diepoxybutane (DEB) is a key metabolite of the important industrial chemical and environmental contaminant, 1,3-butadiene (BD). Although all three optical isomers of DEB, S,S-, R,R-, and meso-DEB, are produced by metabolic processing of BD, S,S-DEB exhibits the most potent genotoxicity and cytotoxicity, followed by R,R- and then meso-DEB. Our previous studies suggested that the observed differences between the biological effects of DEB optical isomers may be structural in their origin. Although S,S- and R,R-DEB produced mainly 1,3 interstrand 1,4-bis-(guan-7-yl)-2,3-butanediol (bis-N7G-BD) cross-links, meso diepoxide induced equal numbers of intrastrand and interstrand bis-N7G-BD lesions. In the present study, the mutagenicity of the three DEB stereoisomers in the supF gene was investigated. We found that S,S-DEB was the most potent mutagen. Interestingly, mutation specificity and mutant spectra were strongly dependent on DEB stereochemistry. Although A:T to T:A transversions were the major form of mutation observed following treatment with each of the three stereoisomers (35-40%), S,S-DEB induced higher numbers of G:C to A:T transitions, whereas R,R-DEB treatment resulted in a greater frequency of G:C to T:A transversions. Our results are consistent with the stereospecific induction of promutagenic nucleobase adducts other than G-G cross-links by DEB stereoisomers. PMID- 17428070 TI - Fragrance compound geraniol forms contact allergens on air exposure. Identification and quantification of oxidation products and effect on skin sensitization. AB - Fragrances are common causes of contact allergy. Geraniol (trans-3,7-dimethyl-2,6 octadiene-1-ol) is an important fragrance terpene. It is considered a weak contact allergen and is used for fragrance allergy screening among consecutive dermatitis patients. Analogous to other monoterpenes studied, such as limonene and linalool, geraniol has the potential to autoxidize on air exposure and form highly allergenic compounds. The aim of the present study was to investigate and propose a mechanism for the autoxidation of geraniol at room temperature. To investigate whether allergenic compounds are formed, the sensitizing potency of geraniol itself, air-exposed geraniol, and its oxidation products was determined using the local lymph node assay in mice. The results obtained show that the allylic alcohol geraniol follows an oxidation pattern different from those of linalool and limonene, which autoxidize forming hydroperoxides as the only primary oxidation products. The autoxidation of geraniol follows two paths, originating from allylic hydrogen abstraction near the two double bonds. From geraniol, hydrogen peroxide is primarily formed together with aldehydes geranial and neral from a hydroxyhydroperoxide. In addition, small amounts of a hydroperoxide are formed, analogous to the formation of the major linalool hydroperoxide. The autoxidation of geraniol greatly influenced the sensitizing effect of geraniol. The oxidized samples had moderate sensitizing capacity, quite different from that of pure geraniol. The hydroperoxide formed is believed to be the major contributor to allergenic activity, together with the aldehydes geranial and neral. On the basis of the present study and previous experience, we recommend that the possibility of autoxidation and the subsequent formation of contact allergenic oxidation products are considered in risk assessments performed on fragrance terpenes. PMID- 17428071 TI - Characterization of fractal particles using acoustics, electroacoustics, light scattering, image analysis, and conductivity. AB - Fractals are aggregates of primary particles organized with a certain symmetry defined essentially by one parameter-a fractal dimension. We have developed a model for the interpretation of acoustic data with respect to particle structure in aggregated fractal particles. We apply this model to the characterization of various properties of a fumed silica, being but one example of a fractal structure. Importantly, our model assumes that there is no liquid flow within the aggregates (no advection). For fractal dimensions of less than 2.5, we find that the size and density of aggregates, computed from the measured acoustic attenuation spectra, are quite independent of the assumed fractal dimension. This aggregate size agrees well with light-scattering measurements. We applied this model to the interpretation of electroacoustic data as well. A combination of electroacoustic and conductivity measurements yields sufficient data for comparing the fractal model of the particle organization with a simple model of the separate primary particles. Conductivity measurements provide information on particle surface conductivity reflected in terms of the Dukhin number (Du). Supporting information for the zeta potential and Du can also be provided by electroacoustic measurements assuming thin double-layer theory. In comparing values of Du from these two measurements, we find that the model of separate solid particles provides much more consistent results than a fractal model with zero advection. To explain this, we first need to explain an apparent contradiction in the acoustic and electroacoustic data for porous particles. Although not important for interpreting acoustic data, advection within the aggregate does turn out to be essential for interpreting electrokinetic and electroacoustic phenomena in dispersions of porous particles. PMID- 17428072 TI - Evidence for the existence of an effective interfacial tension between miscible fluids. 2. Dodecyl acrylate-poly(dodecyl acrylate) in a spinning drop tensiometer. AB - We studied drops of dodecyl acrylate in poly(dodecyl acrylate) (molecular weight of 25,000) in a spinning drop tensiometer to determine whether an effective interfacial tension (EIT) existed between these two miscible fluids. We found convincing evidence. We estimated the mechanical relaxation time from an immiscible analogue (1-propanol and poly(dodecyl acrylate)) and showed that the dodecyl acrylate drops maintained quasi-steady diameters long after this relaxation period. Drops continuously grew in length and became more diffuse, but the width of the transition zone did not grow with t(1/2) as expected from Fick's law although this system had been shown to follow Fick's law in a static configuration (Antrim, D.; Bunton, P.; Lewis, L. L.; Zoltowski, B. D.; Pojman, J. A. J. Phys. Chem. B 2005, 109, 11842-11849). The EIT was determined from Vonnegut's equation, EIT = (Deltarho)omega(2)r(3)/4; both the inner and outer diameters were measured, yielding values of 0.002 and 0.02 mN m(-1), respectively. The EIT was found to be independent of the rotation rate above 6000 rpm and independent of the initial drop volume. The EIT was found to decrease with temperature and increase with the difference in concentration between the monomer drop and polymer-monomer fluid. The square gradient parameter, k, was determined from EIT = k(Deltac(2)/delta), where Deltac is the difference in mole fraction and delta is the width of the transition zone. The square gradient parameter was on the order of 10(-9) N. The square gradient parameter was found to decrease with temperature, to be independent of concentration, and to increase with the molecular weight of the polymer. PMID- 17428073 TI - Reversible condensation of DNA using a redox-active surfactant. AB - We report characterization of aqueous solutions of dilute Lambda phage DNA containing the redox-active surfactant (11-ferrocenylundecyl)trimethylammonium bromide (FTMA) as a function of the oxidation state of the FTMA. FTMA undergoes a reversible one-electron oxidation from a reduced state that forms micelles in aqueous solution to an oxidized state (containing the ferrocenium cation) that does not self-associate in solution. This investigation sought to test the hypothesis that FTMA can be used to achieve reversible control over the conformation of DNA-surfactant complexes in solution. Whereas DNA adopts extended coil conformations in aqueous solutions, our measurements revealed that addition of reduced FTMA (2-5 microM) to aqueous solutions of DNA (5 microM in nucleotide units) resulted in coexistence of extended coils and compact globules in solution. At higher concentrations of reduced FTMA (up to 30 microM), the DNA was present as compact globules only. In contrast, oxidized FTMA had no measurable effect on the conformation of DNA, allowing DNA to maintain an extended coil state up to a concentration of 75 microM oxidized FTMA. We further demonstrate that it is possible to chemically or electrochemically transform the oxidation state of FTMA in preformed complexes of FTMA and DNA, thus achieving in situ control over the conformations of the DNA in solution. These results provide guidance for the design of surfactant systems that permit active control of DNA surfactant interactions. PMID- 17428074 TI - Synergistic assembly of dendrimer-templated platinum catalysts on nitrogen-doped carbon nanotube electrodes for oxygen reduction. AB - A study of the synergistic tuning of nitrogen-doped carbon nanotubes (NCNTs) as support- and size-monodisperse platinum nanoparticles templated from G4-NH2 dendrimers (Pt-DEN's) as catalysts targeted toward oxygen reduction is reported. UV-vis spectroscopy, adsorption isotherms, TGA, TEM, and voltammetry were used to characterize the loading and activity of Pt-DENs immobilized on CNT and NCNT supports. The facile uptake of Pt-DENs was found to be influenced by the number of edge plane sites on the NCNT support with higher adsorption rates observed for NCNTs with increased nitrogen content. Pt-DEN/NCNT composites exhibit high activity with a mass-transport-limited current density and mass activity of 2.3 mA cm(-2) and 0.05 mA g(-1), respectively, for the oxygen reduction reaction (ORR). PMID- 17428075 TI - Dendritic nanoparticles-the impact of ligand cross-linking on nanocore stability. AB - This paper describes the synthesis of gold nanoparticles stabilized by two series of new dendritic disulfide ligands with alkene groups at their peripheries. Intraparticle cross-linking of the alkene groups around the periphery of each nanoparticle was achieved by Grubbs' metathesis. It was demonstrated that cross linking of the organic ligand has no effect on the size or morphology of the inorganic gold core as determined by TEM and UV-vis measurements. However, the introduction of cross-linking at the surface of the ligand enhances the stability of the gold nanocore toward chemical etchant agents (NaCN) and thermal treatment. The impact of cross-linking on nanoparticle stability is greater when the cross linking is closer to the nanoparticle surface (i.e., using lower generation dendritic ligands). Attempts to perform further synthetic transformation on the hybrid materials in order to remove the gold core led to insoluble products composed predominantly of the dendritic ligand. PMID- 17428076 TI - Nanoscale imaging of domains in supported lipid membranes. AB - The formation of domains in supported lipid membranes has been studied extensively as a model for the 2D organization of cell membranes. The compartmentalization of biological membranes to give domains such as cholesterol rich rafts plays an important role in many biological processes. This article summarizes experiments from the author's laboratory in which a combination of atomic force microscopy and near-field scanning optical microscopy is used to probe phase separation in supported monolayers and bilayers as models for membrane rafts. These techniques are used to study binary and ternary lipid mixtures that have gel-phase or liquid-ordered domains that vary in size from tens of nanometers to tens of micrometers, surrounded by a fluid-disordered membrane. Examples are presented in which these models are used to investigate the distribution of glycolipid membrane raft markers and the preference for peptide and protein localization in ordered versus fluid membrane phases. Finally, the enzyme-mediated restructuring of membranes containing liquid-ordered domains provides an in vitro model for the coalescence of membrane rafts to give signaling platforms. Overall, the results demonstrate the importance of using techniques that can probe the nanoscale organization of membranes and of combining techniques that yield complementary information. Furthermore, the ability of supported lipid bilayers to model some aspects of membrane compartmentalization provides an important approach to understanding natural membranes. PMID- 17428077 TI - Surface accessibility of protein post-translational modifications. AB - Protein post-translational modifications are crucial to the function of many proteins. In this study, we have investigated the structural environment of 8378 incidences of 44 types of post-translational modifications with 19 different approaches. We show that modified amino acids likely to be involved in protein protein interactions, such as ester-linked phosphorylation, methylarginine, acetyllysine, sulfotyrosine, hydroxyproline, and hydroxylysine, are clearly surface associated. Other modifications, including O-GlcNAc, phosphohistidine, 4 aspartylphosphate, methyllysine, and ADP-ribosylarginine, are either not surface associated or are in a protein's core. Artifactual modifications were found to be randomly distributed throughout the protein. We discuss how the surface accessibility of post-translational modifications can be important for protein protein interactivity. PMID- 17428078 TI - A multivariate screening strategy for investigating metabolic effects of strenuous physical exercise in human serum. AB - A novel hypothesis-free multivariate screening methodology for the study of human exercise metabolism in blood serum is presented. Serum gas chromatography/time-of flight mass spectrometry (GC/TOFMS) data was processed using hierarchical multivariate curve resolution (H-MCR), and orthogonal partial least-squares discriminant analysis (OPLS-DA) was used to model the systematic variation related to the acute effect of strenuous exercise. Potential metabolic biomarkers were identified using data base comparisons. Extensive validation was carried out including predictive H-MCR, 7-fold full cross-validation, and predictions for the OPLS-DA model, variable permutation for highlighting interesting metabolites, and pairwise t tests for examining the significance of metabolites. The concentration changes of potential biomarkers were verified in the raw GC/TOFMS data. In total, 420 potential metabolites were resolved in the serum samples. On the basis of the relative concentrations of the 420 resolved metabolites, a valid multivariate model for the difference between pre- and post-exercise subjects was obtained. A total of 34 metabolites were highlighted as potential biomarkers, all statistically significant (p < 8.1E-05). As an example, two potential markers were identified as glycerol and asparagine. The concentration changes for these two metabolites were also verified in the raw GC/TOFMS data. The strategy was shown to facilitate interpretation and validation of metabolic interactions in human serum as well as revealing the identity of potential markers for known or novel mechanisms of human exercise physiology. The multivariate way of addressing metabolism studies can help to increase the understanding of the integrative biology behind, as well as unravel new mechanistic explanations in relation to, exercise physiology. PMID- 17428079 TI - Glycoprotein microarrays with multi-lectin detection: unique lectin binding patterns as a tool for classifying normal, chronic pancreatitis and pancreatic cancer sera. AB - Pancreatic cancer is the fourth leading cause of cancer-related death in the United States, with a 5-year survival rate of less than 4%. Effective early detection and screening are currently not available, and tumors are typically diagnosed at a late stage, frequently after metastasis. Existing clinical markers of pancreatic cancer lack specificity, as they are also found in inflammatory diseases of the pancreas and biliary tract. In the work described here, naturally occurring glycoproteins were enriched by using lectin affinity chromatography and then further resolved by nonporous reversed-phase chromatography. Glycoprotein microarrays were then printed and probed with a variety of lectins to screen glycosylation patterns in sera from normal, chronic pancreatitis, and pancreatic cancer patients. Ten normal, 8 chronic pancreatitis, and 6 pancreatic cancer sera were investigated. Data from the glycoprotein microarrays were analyzed using bioinformatics approaches including principal component analysis (PCA) and hierarchical clustering (HC). Both normal and chronic pancreatitis sera were found to cluster close together, although in two distinct groups, whereas pancreatic cancer sera were significantly different from the other two groups. Both sialylation and fucosylation increased as a function of cancer on several proteins including Hemopexin, Kininogen-1, Antithrombin-III, and Haptoglobin related protein, whereas decreased sialylation was detected on plasma protease C1 inhibitor. Target alterations on glycosylations were verified by lectin blotting experiments and peptide mapping experiments using microLC-ESI-TOF. These altered glycan structures may have utility for the differential diagnosis of pancreatic cancer and chronic pancreatitis and identify critical differences between biological samples from patients with different clinical conditions. PMID- 17428080 TI - Relationships between nonadiabatic bridged intramolecular, electrochemical, and electrical electron-transfer processes. AB - Fermi's golden rule is used to develop relationships between rate constants for electron transfer in donor-bridge-acceptor and electrode-bridge-acceptor systems and resistances across metal-bridge-electrode and metal-bridge-tip junctions. Experimental data on electron-transfer rates through alkanethiolate, oligophenylene, and DNA bridges are used to calculate the electronic coupling matrix element per state through these moieties. The formulation is then used to predict the resistance of these bridges between two gold contacts. This approach provides a straightforward method for experimentalists to assess the self consistency between intramolecular electron-transfer rate constants and low-bias resistances measured for molecularly bridged junctions between two metallic contacts. Reported resistances for alkanethiolate bridges vary by a factor of 20, with predicted resistances falling within this range. However, comparisons between carboxylato and directly linked alkanethiolate bridges suggest differences between the coupling at the interface to either the redox center or the gold electrode in such systems. Calculated resistances for oligophenylene bridges are close to those measured experimentally in a similar oligophenylene system. PMID- 17428081 TI - Magnetic interactions in supramolecular N-O...H-C[triple bond]C- type hydrogen bonded nitronylnitroxide radical chains. AB - Two paramagnetic building blocks, 2-(4-ethynyl-1-phenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl (3) and 2-(5-ethynyl-2-pyridyl)-4,4,5,5 tetramethylimidazoline-1-oxyl (4) were synthesized and crystallized. Single crystal X-ray studies of 3 and 4 show the formation of supramolecular head-to tail one-dimensional H-bonded (N-O...H-C[triple bond]C- type) chain structures with O...C distances of 3.181 and 3.155 A, respectively. High-resolution isotropic liquid state (c600 treated patients, satraplatin was generally well tolerated and the most common adverse event was non-cumulative myelosuppression. PMID- 17428105 TI - Management of hepatitis C infection after liver transplantation. AB - Recurrent hepatitis C virus (HCV) infection after orthotopic liver transplantation (OLT) has been associated with progression to cirrhosis in approximately 20% of patients, 5 years postoperatively. Accelerated decompensation has also been noted when compared with cirrhosis in non-transplant patients. Different treatment strategies are available for recurrent HCV infection post-OLT, but efforts are hindered by the modest response rates, poor tolerability and the risk of rejection as well as graft loss. Anti-HCV immunoglobulin therapy to prevent graft infection with HCV has no established role at present but studies are ongoing. Treatment prior to transplantation in patients with decompensated cirrhosis has been evaluated but the results are too preliminary to make firm recommendations. Prophylactic interferon-based antiviral therapy in the early postoperative period to prevent graft infection was shown to have low response rates and high rates of adverse effects. Treatment of established recurrent HCV infection with combination peginterferon (pegylated interferon) and ribavirin is associated with 10-59% sustained virological response and the predictive value of a positive early virological response has been validated in the post-transplant setting. Improvement in inflammatory activity after viral eradication is well established, but fibrosis regression or stabilisation is less predictable and factors such as rejection and biliary complications may still contribute to graft loss. Most studies have initiated therapy at least 6 months postoperatively in order to optimise patient tolerance and enable the addition of ribavirin. The use of adjuvant agents to treat drug induced neutropenia and anaemia in this population is evolving and becoming a crucial part of therapy. Determination of optimal doses of both pegylated interferon and ribavirin, and guidance on when to stop treatment, as well as improving tolerability are important steps in achieving higher response rates and minimising drug toxicity. PMID- 17428106 TI - Montelukast in the treatment of allergic rhinitis: an evidence-based review. AB - Cysteinyl-leukotrienes (CysLTs) are endogenous mediators of inflammation and play an important role in allergic airway disease by stimulating bronchoconstriction, mucus production, mucosal oedema and inflammation, airway infiltration by eosinophils, and dendritic cell maturation that prepares for future allergic response. Montelukast inhibits these actions by blocking type 1 CysLT receptors found on immunocytes, smooth muscle and endothelium in the respiratory mucosa. Initially developed as a treatment for asthma, montelukast has more recently found use in the treatment of allergic rhinitis (AR). We conducted a systematic review of studies that have evaluated montelukast in the treatment of seasonal AR (SAR) and perennial AR (PAR), with and without concomitant asthma. Primary consideration was given to large, randomised, placebo-controlled, double-blind clinical trials in which AR endpoints were assessed and the use of concurrent treatments for AR was excluded. Eight such studies were found in the literature. The primary endpoint in these was daytime nasal symptom severity represented by a composite score derived from individual self-ratings of nasal congestion, rhinorrhoea, nasal pruritus and sneezing. Secondary endpoints have included these individual nasal symptom scores, additional scores for eye, ear and throat symptoms, the impact of rhinitis on quality of sleep, global evaluations of outcome by patients and physicians, and measures of the severity of concomitant asthma. A general outcome was that patients treated with montelukast had significantly greater improvements in their symptoms of SAR and PAR than did patients who were given a placebo. As monotherapy, montelukast exhibited efficacy similar to that of loratadine, but less than that of the intranasally administered corticosteroid fluticasone propionate. The use of montelukast in combination with antihistamines such as loratadine or cetirizine has generally resulted in greater efficacy than when these agents were used alone, and in some studies has produced results comparable with intranasally applied corticosteroids. In patients with AR comorbid with asthma, montelukast treatment has resulted in significant improvements in both, compared with placebo. Montelukast is well tolerated and has a favourable safety profile; adverse events have occurred at similar frequencies in patients taking either montelukast or placebo. Montelukast provides an effective and well tolerated oral treatment for allergic airway inflammation in patients with SAR or PAR without asthma, and in patients in whom AR is comorbid with asthma. PMID- 17428107 TI - Aerosolised antibacterials for the prevention and treatment of hospital-acquired pneumonia. AB - Aerosolised administration of antibacterials remains theoretically attractive for the prevention and treatment of hospital-acquired pneumonia (HAP) because of the ability to generate high drug concentrations at the site of infection. There is renewed interest in this area because of the shortcomings of current therapies and increasing multidrug resistance in Gram-negative organisms. Clinical trials of aerosolised or endotracheally administered antibacterials for HAP prevention have generally been positive; however, early trials were hampered by the development of resistance related to indiscriminate use. More recent trials have shown efficacy at HAP prevention without adverse effects on microflora as a result of more limited usage. However, prophylactic aerosolised antibacterials still need to be studied in large randomised trials before they could enter widespread use. The treatment of HAP with aerosolised antibacterials has mostly been reported in case series without control groups. Both early reports with aminoglycosides and the more recent use of colistin have reported very good response rates; even with organisms such as Pseudomonas aeruginosa and Acinetobacter baumannii. Aerosolised antibacterials were almost always added to intravenous therapy. On the basis of these reports, the current HAP guidelines allow the addition of aerosolised antibacterials in selected patients with multidrug-resistant organisms. This seems to be a reasonable recommendation until large trials are performed. Overall, toxicity was relatively low in the publications reviewed. Aerosolised drug administration in mechanically ventilated patients requires attention to a number of factors in order to maximise drug deposition in the lung. PMID- 17428108 TI - Pharmacological treatments for basal cell carcinoma. AB - Basal cell carcinoma (BCC) is the most common non-melanoma skin cancer, and its incidence continues to rise. Current management options are numerous and focus on tumour eradication while maximising cosmetic and functional capacity. Although surgery continues to be considered the main treatment modality, new pharmacological agents, such as immunomodulators, topical chemotherapeutic agents and photodynamic therapy, have emerged and show promising results. Pharmacological agents offer the potential for lower morbidity and improved tissue preservation compared with surgery and radiotherapy. However, pharmacological treatments possess higher failure rates when compared with surgery, and most studies have investigated only low-risk lesions. Several prospective, randomised, double-blind, vehicle-controlled studies have established the efficacy of imiquimod for superficial BCC. This review summarises the evidence regarding the mechanism, efficacy and safety of pharmacological agents based on the literature from the past 10 years. Experimental treatments that have been successfully utilised in the treatment of BCC are also discussed. Treatment of BCC with other agents, such as tazarotene, glycoalkaloid (BEC-5) cream, cidofovir and calcium dobesilate have been reported, but further studies are needed to ascertain the efficacy and adverse-effect profiles of these treatments. PMID- 17428109 TI - Exenatide: a review of its use in patients with type 2 diabetes mellitus (as an adjunct to metformin and/or a sulfonylurea). AB - Exenatide (Byetta) is a novel, synthetic, incretin mimetic, glucoregulatory peptide approved in the US and Europe for the treatment of patients with type 2 diabetes mellitus who have inadequate glycaemic control despite receiving treatment with maximum tolerated doses of metformin and/or a sulfonylurea. In randomised, controlled, phase III trials and post hoc completer analyses in this patient population, the addition of subcutaneous exenatide twice daily significantly improved glycaemic control and was associated with progressive and significant bodyweight reduction from baseline for up to 2 years. The overall intensity of glycaemic control with exenatide was similar to that achieved with once-daily insulin glargine or twice-daily biphasic insulin aspart. Exenatide was generally well tolerated. Most adverse events were mild to moderate in severity and gastrointestinal in nature. The overall rate of hypoglycaemia was similar to rates observed with placebo (when administered with metformin) and insulin comparators (when administered with metformin and a sulfonylurea). The addition of exenatide to therapy with metformin and a sulfonylurea provided significant improvements in treatment satisfaction and patients' health-related quality of life (HR-QOL). The drug was also cost effective compared with pioglitazone, glibenclamide (glyburide), insulin glargine (all in combination with metformin and/or a sulfonylurea) and metformin alone. Overall, adjunctive therapy with exenatide is a valuable therapeutic option in patients with type 2 diabetes requiring moderate improvements in glycaemic control despite treatment with metformin and/or a sulfonylurea. PMID- 17428110 TI - Extracellular matrix as a strategy for treating chronic wounds. AB - The dermis normally directs all phases of skin wound healing following tissue trauma or disease. However, in chronic wounds, the dermal matrix is insufficient to stimulate healing and assistance by external factors is needed for wound closure. Although the concept of the extracellular matrix directing wound healing is not new, ideas about how best to provide the extracellular matrix components required to 'jump-start' the healing process are still evolving. Historically, these strategies have included use of enzyme-inhibiting dressing materials, which bind matrix metalloproteinases and remove them from the chronic wound environment, or direct application of purified growth factors to stimulate fibroblast activity and deposition of neo-matrix. More recently, the application of a structurally intact, biochemically complex extracellular matrix, designed to provide the critical extracellular components of the dermis in a single application, has allowed for the reconstruction of new, healthy tissue and restoration of tissue integrity in the previously chronic wound. This review focuses on this third mechanism as an emerging tactic in effective wound repair. Intact extracellular matrix can quickly, easily, and effectively provide key extracellular components of the dermis necessary to direct the healing response and allow for the proliferation of new, healthy tissue. Its application may promote the healing of wounds that have been refractory to other, more conventional treatment strategies, and may eventually show utility when used earlier in wound healing treatment with the goal of preventing wounds from reaching a truly chronic, nonresponsive state. PMID- 17428111 TI - The role of anti-tumor necrosis factor-alpha therapy in Pyoderma gangrenosum associated with inflammatory bowel disease. AB - Pyoderma gangrenosum (PG) is an ulcerative neutrophilic dermatosis seen in 1-5% of patients with inflammatory bowel disease (IBD). The pathogenesis of PG remains unclear, but may be related to abnormal T-cell responses and production of tumor necrosis factor (TNF)-alpha, a powerful proinflammatory cytokine. Although their use is not supported by appropriately controlled trials, corticosteroids and systemic immunosuppressants are the classical cornerstones of treatment of PG, against which they have a nonspecific effect. Successful curative or symptomatic treatment of associated disorders may lead to an improvement in PG. A new era for the management of chronic inflammatory disease began with the advent of biotherapies and particularly anti-TNFalpha therapy, which allows for a specific intervention in the immune cascade. Anti-TNFalpha therapy has improved and broadened the therapeutic options for IBD and, therefore, has brought new perspectives to management of the extra-intestinal manifestations of this disorder, including PG. To date, infliximab, etanercept, and adalimumab have been used in the treatment of PG. Published data have demonstrated that infliximab is highly effective in the treatment of PG, whether associated with IBD or not. This treatment is generally well tolerated, even as long-term therapy. However, rare and serious complications have been reported. Although infliximab is a costly drug, its use should be considered for patients with PG and particularly with corticosteroid-refractory PG associated with IBD. Additional comparative long term studies are needed to determine the long-term efficacy and safety of anti TNFalpha therapy and define its role in the management of PG, with or without accompanying IBD. PMID- 17428112 TI - Perineal herpes simplex infection in bedridden geriatric patients. AB - BACKGROUND: Herpes simplex virus (HSV) lesions are prone to reactivation and recurrence in response to various local or systemic triggering factors. OBJECTIVE: To study the characteristics of five bedridden geriatric patients who presented with herpetic recurrences on the buttocks, gluteal cleft, and perianal region during hospitalization. METHODS: Data were gathered regarding age, gender, reason for hospitalization, localization of lesions, clinical presentation, previous clinical diagnosis and topical treatments, immune status and immunosuppressant drug intake, as well as prior history of labial or genital herpes. A skin biopsy was taken for histologic examination and immunohistochemical viral identification. Viral culture and viral serology were performed and data regarding antiviral therapy were recorded. RESULTS: The five patients (three women, two men) were aged >80 years and hospitalized for either severe drug-induced renal insufficiency (one case), severe pneumonia (two cases), or stroke causing restricted mobility (two cases). Numerous well demarcated, painful ulcerations developed in the perianal region of these patients, and one patient also presented with some vesicular lesions. The lesions had been confused with mycotic and/or bacterial infections for 10-14 days. No inguinal lymphadenopathies were present and there was no fever. None of the patients had a previous history of recurrent labial or genital HSV infections or HIV infection. Histology was suggestive of HSV infection in two of five patients. Immunohistochemistry identified HSV type I (three patients) and HSV type II (two patients) infections. Viral culture with immunofluorescence viral identification revealed HSV type I in one of the four patients in whom a swab for viral culture was taken. Serology revealed past HSV infection. All lesions cured gradually after 10-14 days of intravenous acyclovir (aciclovir) treatment. CONCLUSION: Herpetic lesions of the perineal region represent a rare complication in bedridden geriatric patients in the absence of a previous history of HSV infections at the same site. Common traits of patients with this condition were the presence of numerous ulcerated lesions, prolonged time course, and confinement to bed. The latter probably modifies the skin condition, which triggers viral reactivation and favors cutaneous extension of the infection. Complementary diagnostic methods for viral detection and identification are mandatory. PMID- 17428113 TI - Efficacy and safety of cyclophosphamide, azathioprine, and cyclosporine (ciclosporin) as adjuvant drugs in pemphigus vulgaris. AB - BACKGROUND: Pemphigus vulgaris is a potentially life-threatening, autoimmune bullous disease of the skin and mucous membranes. Most commonly, the disease is treated with prednisone in combination with an immunosuppressant agent, frequently referred to as adjuvant drug. However, there is no consensus regarding the first-choice adjuvant drug for the treatment of pemphigus vulgaris or the recommended dosage. OBJECTIVE: To evaluate the efficacy and safety of prednisone as monotherapy and in combination with the three most popular adjuvant agents - azathioprine, cyclosporine (ciclosporin), and cyclophosphamide - in the treatment of pemphigus vulgaris. METHODS: This was a retrospective study with a follow-up of 7-21 years. The study was conducted in an academic hospital with an outpatient division for patients with bullous diseases. A total of 101 patients with moderate-to-severe mucocutaneous pemphigus vulgaris were included in the study. For assessment of disease severity a 'pemphigus score,' based on the percentage of involved skin or oral mucous membranes, was developed. At treatment initiation the average pemphigus score was comparable in all treated groups of patients. Four treatment regimens were evaluated: oral prednisone at an initial dose of 100mg (1.1-1.5 mg/kg) per day as monotherapy, and prednisone combined with adjuvant drugs, i.e. oral azathioprine at a dose of 100mg (1.1-1.5 mg/kg) per day; cyclosporine (ciclosporin) at a dose of 2.5-3 mg/kg/day; or cyclophosphamide at a dose of 100mg (1.1-1.5 mg/kg) per day. The main outcome measures were average time to clinical remission, average time to immunologic remission (non detectable circulating pemphigus vulgaris antibodies), proportion of patients who remained free of clinical relapse within 5 years after discontinuation of therapy, time from treatment discontinuation until first relapse, and incidence of adverse effects. RESULTS: The average (+/- SD) time to clinical remission was 7.2 +/- 13.1 months in patients who received prednisone monotherapy, 6.8 +/- 10.5 months in patients receiving additional azathioprine, 8.1 +/- 11.8 months in the cyclosporine group, and 4.9 +/- 6.9 months (which was significantly shorter than all other treatment groups, p < 0.05) in patients receiving cyclophosphamide. The average (+/- SD) times to immunologic remission were 33 +/- 27 months, 28 +/- 24 months, 30 +/- 21 months, and 23 +/- 17 months for prednisone monotherapy, azathioprine, cyclosporine, and cyclophosphamide, respectively. The proportions of patients who remained free of clinical relapse within 5 years after discontinuation of therapy were 55%, 50%, 43%, and 69% for prednisone monotherapy, azathioprine, cyclosporine, and cyclophosphamide, respectively. In patents who experienced relapse, the average (+/- SD) time from treatment discontinuation to clinical relapse was 10.50 +/- 6.86 months in patients receiving prednisone monotherapy, 16.40 +/- 17.36 months in the azathioprine group, 12.44 +/- 6.48 months in the cyclosporine group, and 21.16 +/- 20.13 months in the cyclophosphamide group. The safety profiles of all treatment regimens were comparable. CONCLUSION: Oral prednisone with cyclophosphamide is the most effective treatment for pemphigus vulgaris. All therapy regimens had a similar safety profile. In our opinion, cyclophosphamide at a dose of 1.1-1.5 mg/kg/day should be the adjuvant drug of choice in the treatment of moderate-to severe pemphigus vulgaris. PMID- 17428114 TI - Placebo response in two long-term randomized psoriasis studies that were negative for rosiglitazone. AB - BACKGROUND: Previous research has suggested that the thiazolidinedione rosiglitazone may possess anti-psoriatic activity. OBJECTIVE: To compare the efficacy and safety of rosiglitazone with that of placebo in the treatment of chronic plaque psoriasis. METHODS: Two large-scale, randomized, double-blind, multicenter studies (study A, n = 1563; study B, n = 1032) were conducted over 52 weeks (plus optional 44 weeks safety extension) in an outpatient setting. The subjects (aged 18-75 years) had moderate-to-severe chronic plaque psoriasis affecting >or=10% body surface area (BSA) with plaques of any elevation above normal-appearing skin (or >or=6% BSA involvement with marked elevation) and had not used phototherapy during the previous month or thiazolidinediones within the previous 3 months. Rosiglitazone was administered as 2, 4, or 8 mg tablets once daily. The main outcome measure was the proportion of subjects achieving >or=75% improvement from baseline in the Psoriasis Area and Severity Index (PASI 75) at week 26. RESULTS: Rosiglitazone was well tolerated but no more effective than placebo for moderate-to-severe chronic plaque psoriasis. However, there was a large placebo response unrelated to concomitant rescue medication. Interestingly, subjects had been advised to expect a long period before onset of action. At week 26 and across both studies for subjects receiving placebo, the PASI 75 was 9% (48/506) and the PASI 50 (proportion of subjects who achieved at least 50% improvement from baseline) was 27% (135/506). In addition, few subjects withdrew from placebo or rosiglitazone treatment because of 'lack of efficacy' and the majority persisted in the year-long study. CONCLUSION: While these large-scale, robust studies demonstrated that rosiglitazone is not active in psoriasis, they also showed that for a large proportion of subjects receiving placebo, the expectation of a successful treatment, the favorable adverse effect profile of the drug, and the supportive environment of a clinical study conferred beneficial effects. These results may have implications for the design of future placebo controlled studies in patients with psoriasis. PMID- 17428115 TI - Infantile scurvy: an old diagnosis revisited with a modern dietary twist. AB - Ascorbic acid (vitamin C) is necessary for the formation of collagen, reducing free radicals, and aiding in iron absorption. Scurvy, a disease of dietary ascorbic acid deficiency, is uncommon today. Indeed, implementation of dietary recommendations largely eradicated infantile scurvy in the US in the early 1900s. We present a case of an otherwise healthy 2-year-old Caucasian girl who presented with refusal to walk secondary to pain in her lower extremities, generalized irritability, sleep disturbance, and malaise. The girl's parents described feeding the patient an organic diet recommended by the Church of Scientology that included a boiled mixture of organic whole milk, barley, and corn syrup devoid of fruits and vegetables. Physical examination revealed pale, bloated skin with edematous, violaceous gums and loosening of a few of her teeth. Dermatologic findings included xerosis, multiple scattered ecchymoses of the extremities, and perifollicular hemorrhage. Laboratory and radiographic evaluation confirmed the diagnosis of scurvy. The patient showed dramatic improvement after only 3 days of treatment with oral ascorbic acid and significant dietary modification. In this case report, we revisit the old diagnosis of scurvy with a modern dietary twist secondary to religious practices. This case highlights the importance of taking a detailed dietary history when evaluating diseases involving the skin. PMID- 17428116 TI - Stevens-Johnson syndrome due to concomitant use of lamotrigine and valproic acid. AB - Stevens-Johnson syndrome (SJS) is a rare but life-threatening acute mucocutaneous hypersensitivity reaction, usually related to drugs. Severe cutaneous adverse effects such as SJS and toxic epidermal necrolysis can arise during treatment with antiepileptic drugs (AEDs). A 23-year-old female patient with idiopathic epilepsy was referred to the emergency service with fever, oral and genital mucosal lesions, generalized rash, and weakness. She had been taking carbamazepine, valproic acid, and lamotrigine until 1 week prior to referral. Following consultations with a range of clinicians, the patient was diagnosed with SJS related to lamotrigine on the basis of her history and physical findings, and on consideration of current consensus definitions of this condition. The patient was followed up and treated in the emergency intensive care unit with intravenous fluids, antibacterial therapy, and methylprednisolone. After 18 days, the patient was considered to have made a sufficient recovery and was discharged. Patients who undergo therapy with AEDs, particularly new users of these agents, should be informed of and monitored for possible systemic and cutaneous adverse effects of AEDs. Polytherapy should be avoided for as long as possible as this increases the frequency of adverse effects, and treatment should be discontinued if any rash appears. PMID- 17428117 TI - Punctate palmoplantar keratoderma (Brauer-Buschke-Fischer syndrome). AB - Punctate palmoplantar keratoderma (PPPK) is a rare entity with an estimated prevalence rate of about 1.17 per 100,000. The exact etiology of the disorder is not known but a dual influence of genetic and environmental factors may trigger the disease. We report the case of a 70-year-old male patient with punctate palmoplantar keratodermic lesions for more than 40 years. Histopathologic examination revealed a hyperkeratotic epidermis without columns of parakeratosis or elastorhexis. On electron microscopy, the basal cells of the epidermis were found to have enlarged nucleoli and abundant tonofilaments, with keratohyalin like granules confined to the upper part of the stratum spinosum, findings that were consistent with PPPK. Topical keratolytic agents were used with little success. Patients with PPPK and their next of kin should be investigated for possible associated malignancies. PMID- 17428118 TI - Cutaneous spherulosis (myospherulosis). PMID- 17428119 TI - Etanercept for the management of oral lichen planus. PMID- 17428120 TI - Towards a truly biomimetic olfactory microsystem: an artificial olfactory mucosa. AB - Today, the capability of the human olfactory system is still, in many ways, superior to that of the electronic nose. Although electronic noses are often compared with their biological counterpart, they neither mimic its neural architecture nor achieve its discriminating performance. Experimental studies on the mammalian olfactory system suggest that the nasal cavity, comprising of the mucous layer and the olfactory epithelium, performs a degree of chromatographic separation of complex mixtures. Thus receptor cells distributed beneath the mucous layer provide both spatial and temporal chemosensory information. Here we report on the development of an artificial olfactory microsystem that replicates this basic structure. This contains an integrated channel to emulate the nasal cavity and coated with a polymer to mimic the partitioning mucous layer, which is positioned directly over a sensor array. Our system employs an 80 element chemoresistive microsensor array with carbon black/polymer odour-sensitive films combined with a microfluidic package fabricated by micro-stereolithography. Results show that this biomimetic system generates both spatial and temporal odorant signals, with a temporal chemical retention period of up to 170 s. Data analysis has revealed improvements in its ability to discriminate between two simple odours and a set of complex odours. We believe such emulation of the olfactory system can lead to improved odour discrimination within the field of electronic noses. PMID- 17428121 TI - Nanostructuring of heme-proteins for biodevice applications. AB - Proteins represent versatile building blocks for the realisation of nanostructured materials to be applied in the nanobiotechnological field. The Langmuir-Blodgett (LB) technique was utilised as a means to develop nanobiodevices based on protein molecules. Namely, engineered Cytochrome P450 thin films were fabricated and characterised. The possibility to employ LB-based protein structures to use in biosensors has been exploited. The characterisation process was performed in order to verify the best working parameters. As a first step' the protein films were studied at the air-water interface and then transferred into a solid support for further characterisation. The films were characterised by different techniques such as: UV-vis spectroscopy, nanogravimetry, atomic force microscopy and biochemical assays. The results showed that it was possible to form active cytochrome P450s nanostructures by the LB technique. PMID- 17428122 TI - Understanding social and professional integration as an adjustment process: contribution to a theory of coadaptation. AB - This qualitative research was designed to understand how the social integration process in the workplace unfolds with adults who have intellectual disabilities. Participant observation and interviews permitted an in-depth examination of 10 integration situations within a company, with regard for the actions and perceptions of the stakeholders. Analyses revealed various intertwined processes, in which the focus persons and their colleagues together sought an optimal adjustment, or coadaptation, with support from a job coach. The study of these interactive processes led to the emergence of a theory of coadaptation, a process that defines and builds social and professional integration. This theoretical development revisits the interactive process within the socioecological perspective of disability and the concept of support and its assessment. PMID- 17428123 TI - Cell phone use by adults with intellectual disabilities. AB - Although cell phone use has grown dramatically, there is a gap in cell phone access between people with disabilities and the general public. The importance of cell phone use among people with intellectual disabilities and studies about use of cell phones by adults with intellectual disabilities was described. Our goal was to determine the extent and scope of cell phone use by 83 adults with intellectual disabilities, reasons for nonuse, and factors affecting use. Results suggest a gap in the use of cell phone technology between people with intellectual disabilities and the general population. When used, they are employed primarily for emergencies, storing telephone numbers, and day-to-day communication. Chief reasons for nonuse include cost, perception of not needing one, and lack of accessibility. PMID- 17428124 TI - Developmental concept of idiocy. AB - In dominant definitions of mental retardation, researchers have insisted on the diagnosis being restricted to conditions manifested during the developmental period. However, even in the 19th century, this was only one of several conceptual options, some of which did not exclude adult brain injury or dementia. Events in the 19th and early 20th centuries, particularly with the growth of institutions, scientific study, and, later, intelligence testing led to these other options being excluded. Here, I discuss the proposal that current definitions are highly contingent on factors that are neither essential nor necessary. Although not arguing for any specific changes to current definitions, I do argue that theoretical options should be kept open and that dominant ones should be questioned. PMID- 17428125 TI - Project MED: effects of a Medication EDucation booklet series for individuals with intellectual disabilities. AB - We developed eight heavily illustrated booklets covering patients' rights and responsibilities, antiepileptic medicines, and most psychotropic medicines. The language level was very basic but covered a wide range of information. We distributed free copies of the booklets, together with standardized questionnaires, to consumers with and without intellectual disabilities or other developmental disabilities; 604 questionnaires were returned. The majority of consumers indicated that they read the booklets, learned more about their rights/responsibilities and the medicines described, and found the booklets helpful. Consumers with intellectual disability experienced more difficulty than "average" consumers in understanding the materials, but satisfaction and understanding were reportedly high overall. Female and minority respondents indicated somewhat higher satisfaction with the booklets than did their counterparts. PMID- 17428126 TI - Including students with moderate and severe intellectual disabilities in school extracurricular and community recreation activities. AB - We conducted an online statewide survey of teachers of students with moderate and severe intellectual disabilities to determine the extent to which their students were included in school extracurricular and community recreation activities. For the 252 teacher respondents who indicated that their primary caseload consisted of students with significant intellectual disabilities, we report the numbers of students participating in school and community activities and the primary type of support students required to participate in each activity. Finally, we identify implications for practitioners who want to increase the participation of students with significant disabilities in school and community activities. PMID- 17428128 TI - Perspective on quality of life in dementia care. PMID- 17428129 TI - Police officers and disability: perceptions and attitudes. PMID- 17428133 TI - Effects of videorecorded interactions and counseling for teachers on their responses to preschoolers with intellectual impairments. AB - Effects of an intervention and video interaction counseling with teachers of 78 children from school and centers were investigated. The study involved a quasi experimental pretest-intervention-posttest control group design. In Condition 1 teachers received the counseling for 12 weeks, followed by withdrawal of this intervention for weeks. In Condition 2 the intervention was in reversed order. The students' competent behavior was assessed by rating their response to social support during engagement in a standardized task, their performance of standardized tasks of cognitive skills, and ratings of their social skills. Results revealed differences in favor of the participants in the two video interaction counseling research conditions on measures of social support and cognitive competence. PMID- 17428134 TI - The renaming of mental retardation: understanding the change to the term intellectual disability. PMID- 17428135 TI - Taking risks: creating powerful dispositions for professionals working with people who have developmental disabilities and their families. PMID- 17428136 TI - End-of-life care challenges for persons with intellectual disability and dementia: making decisions about tube feeding. PMID- 17428137 TI - From whence came mental retardation? Asking why while saying goodbye. PMID- 17428138 TI - Counter the mistreatments for autism with professional integrity. PMID- 17428139 TI - The fruitlessness of trying to convey competencies to people while ignoring their image. PMID- 17428140 TI - Gisli Gudjonsson's penchant for corroboration. PMID- 17428141 TI - State institution populations in 2006 less than 80% of 40 years earlier; "real dollar" per person costs more than 10 times greater. PMID- 17428143 TI - Attitudes of juvenile justice staff towards intellectual, psychiatric, and physical disabilities. AB - Juvenile justice employees' attitudes towards diverse impairments and their ideological and demographic predictors were examined. Supporting theories are that attitudes are comprised of ideological constructs and demographic variables and that there is a hierarchy of disability present in attitudes towards diverse disabilities. A cross-sectional design with quantitative measures was used. Results indicate that attitudes towards different types of disabilities vary considerably and are hierarchical in nature and that independent variables of demographics, social desirability in responding, and ideological constructs of social dominance orientation, normativism, and humanism have some effects on attitude outcomes, but not universally. PMID- 17428144 TI - Factors that impact resuscitation preferences for young people with severe developmental disabilities. AB - A cross-sectional descriptive study was performed to evaluate resuscitation decisions and factors that impact these choices for young people with severe developmental disabilities residing in a skilled nursing facility. Decision makers were provided with information to clarify resuscitation preferences. Parents/guardians of 30 of the 67 residents also completed a survey. A significant number of decision-makers changed their resuscitation preference to DNR after detailed explanations were provided. Survey results suggest that interpersonal relationships, such as those with family members, religious leader, and physician, were more influential for families who chose full resuscitation compared to those with DNR preferences. Factors such as perception of quality of life and medical condition of the individuals with developmental disabilities were not significantly different between these two groups. PMID- 17428145 TI - Attitudes of mothers towards their child with Down syndrome before and after the introduction of prenatal diagnosis. AB - In 1970, before the introduction of prenatal diagnosis of chromosome anomalies, an unpublished questionnaire study concerning the social and emotional situation of mothers of children with Down syndrome was conducted in southern Germany. To assess the psychosocial impact of the availability of prenatal diagnosis on parents of genetically handicapped children, we re-evaluated and repeated the 1970 study over 30 years later. Although mothers' feelings of guilt for having a child with disabilities remained on a low level, today's mothers have a stronger feeling of being involuntarily segregated in society. On the other hand, they more often experience support and respect from outside, particularly through self support groups; moreover, tendencies of active withdrawal from social life have decreased. PMID- 17428146 TI - Metabolism and pharmacokinetics in children and the elderly. AB - In drug development, clinical trials are commonly carried out in either healthy volunteers or carefully selected patients. However, it has been recognised for a long time that both extremes of age, children and elderly, display a number of important metabolic and pharmacokinetic differences as compared with adults. This also has important consequences as to the toxic reactions and risk assessment of chemicals. What is known, what is not known and what should we do to improve the situation? PMID- 17428147 TI - Stereoselectivity in drug metabolism. AB - Many chiral drugs are used as their racemic mixtures in clinical practice. Two enantiomers of a chiral drug generally differ in pharmacodynamic and/or pharmacokinetic properties as a consequence of the stereoselective interaction with optically active biological macromolecules. Thus, a stereospecific assay to discriminate between enantiomers is required in order to relate plasma concentrations to pharmacological effect of a chiral drug. Stereoselective metabolism of drugs is most commonly the major contributing factor to stereoselectivity in pharmacokinetics. Metabolizing enzymes often display a preference for one enantiomer of a chiral drug over the other, resulting in enantioselectivity. The structural characteristics of enzymes dictate the enantiomeric discrimination associated with the metabolism of chiral drugs. The stereoselectivity can, therefore, be viewed as the physical property characteristic that phenotypes the enzyme. This review provides a comprehensive appraisal of stereochemical aspects of drug metabolism (i.e., enantioselective metabolism and first-pass effect, enzyme-selective inhibition or induction and drug interaction, species differences and polymorphic metabolism). PMID- 17428148 TI - Amino acid conjugation: contribution to the metabolism and toxicity of xenobiotic carboxylic acids. AB - Despite being the first conjugation reaction demonstrated in humans, amino acid conjugation as a route of metabolism of xenobiotic carboxylic acids is not well characterised. This is principally due to the small number and limited structural diversity of xenobiotic substrates for amino acid conjugation. Unlike CYP and uridine 5'-diphosphate glucuronosyltransferase, which are localised in the endoplasmic reticulum, the enzymes of amino acid conjugation reside in mitochondria. Unique among drug metabolism pathways, amino acid conjugation involves initial formation of a xenobiotic acyl-CoA thioester that is then conjugated principally with glycine in humans. However, formation of the xenobiotic acyl-CoA thioester does not always infer subsequent amino acid conjugation. Evidence is presented that in the absence of glycine conjugation substrates that form acyl-CoA thioesters perturb mitochondrial function. This review discusses literature on the enzymes involved and the concept that xenobiotic substrate selectivity provides a barrier to protect the metabolic integrity of the mitochondria. PMID- 17428149 TI - Arylamine N-acetyltransferases. AB - Arylamine N-acetyltransferases (NATs), known as drug- and carcinogen-metabolising enzymes, have had historic roles in cellular metabolism, carcinogenesis and pharmacogenetics, including epidemiological studies of disease susceptibility. NAT research in the past 5 years builds on that history and additionally paves the way for establishing the following new concepts in biology and opportunities in drug discovery: i) NAT polymorphisms can be used as tools in molecular anthropology to study human evolution; ii) tracing NAT protein synthesis and degradation within cells is providing insight into protein folding in cell biology; iii) studies on control of NAT gene expression may help to understand the increase in the human NAT isoenzyme, NAT1, in breast cancer; iv) a NAT homologue in mycobacteria plays an essential role in cell-wall synthesis and mycobacterial survival inside host macrophage, thus identifying a novel biochemical pathway; v) transgenic mice, with genetic modifications of all Nat genes, provide in vivo tools for drug metabolism; and vi) structures of NAT isoenzymes provide essential in silico tools for drug discovery. PMID- 17428150 TI - Role of signalling systems in the effects of dietary factors on the expression of mammalian CYPs. AB - Changes in mammalian diets alter the hepatic expression of CYP drug-metabolising enzymes and endobiotic oxidases. Thus, dietary constituents may significantly influence the duration of action of chemicals in tissues. Recent improvements in the mechanistic information on the regulation of constitutive and inducible expression of CYPs has facilitated our understanding as to how dietary factors modulate expression. Altered regulation appears to occur either by direct activation of transcription factors or by indirect modulation of signal transduction pathways. For example, dietary lipid directly activates PPAR-alpha, or other nuclear hormone receptors, to elicit CYP induction, and vitamin A deficiency downregulates the growth hormone-responsive CYP2C11 by perturbing Janus kinase-signal transducers and activators of transcription signalling. This article focuses on the present understanding of the regulation of CYP genes by dietary nutrients. PMID- 17428151 TI - Drug interactions between proton pump inhibitors and antiretroviral drugs. AB - Gastroesophageal reflux disease and other related symptoms are common comorbidities for HIV-infected patients. Therefore, concurrent use of proton pump inhibitors and antiretrovirals is a common practice in the clinical setting. However, this practice may alter antiretroviral pharmacokinetics and lead to treatment failure due to inadequate drug exposure. Clinician awareness of these complex interactions is essential for optimal HIV management. This review summarizes the current knowledge on the interactions between proton pump inhibitors and antiretrovirals, and makes recommendations for the coadministration of proton pump inhibitors. PMID- 17428152 TI - Toxicity as a result of immunostimulation by biologics. AB - The immune system has evolved highly effective mechanisms of surveillance and defense against foreign pathogens, and is also thought to act in surveillance and suppression of cancer. Thus, a predominant goal of immune system-based therapies is to normalize or enhance the host immune response in the areas of infectious disease and oncology. This review considers general approaches used for therapeutic immunostimulation, alterations in immune response mechanisms that occur with these treatments and the syndromes that commonly arise as a result of these changes. Because nonclinical studies of these therapies are conducted in animal models as the basis for predicting potential human toxicities, this review also considers the value of nonclinical testing to predict human toxicity. PMID- 17428153 TI - Whole-body physiologically based pharmacokinetic models. AB - This review summarizes the most recent developments in and applications of physiologically based pharmacokinetic (PBPK) modeling methodology originating from both the pharmaceutical and environmental toxicology areas. It focuses on works published in the last 5 years, although older seminal papers have also been referenced. After a brief introduction to the field and several essential definitions, the main body of the text is structured to follow the major steps of a typical PBPK modeling exercise. Various applications of the methodology are briefly described. The major future trends and perspectives are outlined. The main conclusion from the review of the available literature is that PBPK modeling, despite its obvious potential and recent incremental developments, has not taken the place it deserves, especially in pharmaceutical and drug development sciences. PMID- 17428154 TI - Therapeutic drug monitoring of mycophenolic acid: does it improve patient outcome? AB - Treatment with the immunosuppressive agent mycophenolate mofetil (MMF) decreases the risk of rejection after renal transplantation and improves graft survival compared with azathioprine. The exposure to the active metabolite mycophenolic acid (MPA) is correlated to the risk of developing acute rejection. The interpatient variability in exposure of MPA is wide relative to the proposed therapeutic window of the MPA AUC(0 12) (30 - 60 mg.h/l). The pharmacokinetics of MPA are influenced by patient characteristics such as gender, time after transplantation, serum albumin concentration, renal function, comedication and pharmacogenetic factors. Therapeutic drug monitoring is likely to reduce inter patient variability. Limited sampling strategies are used to predict the full AUC(0 12). Three prospective randomised studies compared concentration controlled MMF therapy to a fixed-dose regimen. Preliminary outcomes of these studies showed conflicting results and longer follow up is needed to further clarify the role of therapeutic drug monitoring in increasing the therapeutic potential of MMF. PMID- 17428155 TI - The pharmacology and clinical use of caspofungin. AB - Caspofungin was the first echinocandin to be licensed for the treatment of invasive fungal infections. Caspofungin has in vitro and in vivo activity against Candida spp. and Aspergillus spp., which constitute the majority of medically important opportunistic fungal pathogens. Caspofungin inhibits the synthesis of the 1,3-beta-glucan, with resultant osmotic instability and lysis. The pharmacology of caspofungin is relatively complex. Trafficking of drug into tissues is an important determinant of the shape of the concentration-time relationship. Caspofungin has demonstrated efficacy in experimental models of invasive candidiasis and aspergillosis, which reflect its activity in the treatment of oropharyngeal, esophageal and disseminated candidiasis, as well as salvage therapy for patients with invasive aspergillosis. PMID- 17428156 TI - Current strategies with high-dose tirofiban. AB - The glycoprotein (GP) IIb/IIIa receptor is a platelet-specific adhesion receptor that mediates the formation of platelet aggregates. Pharmacologic blockade of the receptor is associated with a reduction in major cardiovascular adverse events after percutaneous coronary interventions and in the setting of acute coronary syndromes. Three intravenous GP IIb/IIIa receptor inhibitors are available: abciximab, tirofiban and eptifibatide. Tirofiban is a small, synthetic non peptide, competitive GP IIb/IIIa antagonist with high specificity and high affinity for the GP IIb/IIIa receptor. In a head-to-head comparison, tirofiban 10 microg/kg bolus followed by a 0.15-microg/kg/min infusion was found to be inferior to standard dose of abciximab in patients undergoing percutaneous coronary intervention. Insufficient platelet inhibition with low-dose tirofiban may likely explain these results. Subsequently, a high-bolus dose of tirofiban (25 microg/kg bolus) followed by standard infusion was tested and evidence suggest that in this dosing tirofiban may be as effective as abciximab and have a comparable safety profile. Therefore, high-bolus dose tirofiban may be an appealing and cost-effective alternative to abciximab. However, further testing is warranted given the short follow up and limited statistical power of the available data. PMID- 17428157 TI - Oxaliplatin in the treatment of colorectal cancer. AB - Significant advances in the treatment of colorectal cancer have been observed over the past several years. With the introduction of oxaliplatin combined with infusional 5-fluorouracil and leucovorin, survival for patients with metastatic colorectal cancer has nearly doubled. The incorporation of biologic agents that target angiogenesis (bevacizumab) and tumor growth pathways (cetuximab, panitumimab) extends survival even further, in addition to increasing response rates in patients with metastatic disease. The benefit of these newer drugs is also being realized in the adjuvant setting, where the addition of oxaliplatin to infusional 5-fluorouracil and leucovorin has led to improvements in 3-year disease-free survival. Future challenges with the use of oxaliplatin include defining strategies to optimize its use while avoiding treatment-limiting neurotoxicity and identification of markers predictive of response. PMID- 17428158 TI - High-throughput in vitro hemotoxicity testing and in vitro cross-platform comparative toxicity. AB - For decades, hemotoxicity has been considered as simply changes in peripheral blood parameters or morphological changes observed in the bone marrow as a result of drug administration. The effects are actually the result of the drug acting at a much earlier level in the blood-forming system, usually at the level of the lympho-hematopoietic stem cell or its immediate differentiating progeny. To detect these early cellular responses, highly sensitive, non-subjective and fully standardized in vitro high-throughput testing platforms have been developed that detect changes in intracellular ATP concentrations which are directly proportional to and predict the proliferative/cytotoxic response of different cell populations. PMID- 17428159 TI - Curability of lung cancer. PMID- 17428161 TI - Cetuximab plus irinotecan in refractory colorectal cancer patients. AB - Since the mid-1980s, it has been known that the epidermal growth factor receptor, an immunoglobulin G cytoplasmic membrane protein and member of the type I subfamily of tyrosine kinase receptors, plays a key role in tumor growth and metastasis. Cetuximab is the first immunoglobulin G1 monoclonal antibody that blocks the epidermal growth factor receptor, resulting in an inhibition of tumor growth, angiogenesis, tumor spread and metastasis. Cetuximab is active in heavily pretreated colorectal cancer, as a single agent or in combination with irinotecan, a conclusion that is strongly suggested by the Bowel Oncology with Cetuximab Antibody (BOND) study in terms of response rate (11% monotherapy vs 23% in combination; p < 0.007) and time to progression (1.5 vs 4.1 months; p < 0.001). The main adverse event related to cetuximab is skin reaction, the intensity of which is correlated with efficacy. The Monoclonal Antibody Erbitux in a European Pre-License (MABEL) study has confirmed these results, recently generating a median survival of 9.2 months, comparable to 8.6 months achieved in the BOND study. Cetuximab has shown efficacy (with or without irinotecan) as second-line treatment in patients who have failed irinotecan in combination with bevacizumab, an antivascular endothelial growth factor monoclonal antibody. The combination is currently being tested in the first-line management of advanced colorectal cancer, as well as in the adjuvant setting. The evaluation of other combinations either with oxaliplatin or capecitabine, is ongoing, with preliminary reports of promising activity. PMID- 17428162 TI - Paclitaxel poliglumex (PPX, CT-2103): macromolecular medicine for advanced non small-cell lung cancer. AB - Taxanes, including paclitaxel and docetaxel, are widely used cytotoxic agents for the treatment of solid tumors. Paclitaxel, a small, hydrophobic agent, binds extensively to plasma proteins, and its pharmacokinetic profile is characterized by a short plasma elimination half-life with a broad tissue distribution. These unfavorable pharmacokinetic characteristics are associated with limited tumor exposure and high systemic exposure, reducing the therapeutic index of paclitaxel. Paclitaxel poliglumex (PPX, CT-2103), a polymer-drug conjugate of paclitaxel and poly-L-glutamic acid, was designed to enhance the therapeutic index of paclitaxel by improving its pharmacokinetic profile, and to provide a water-soluble alternative to the standard paclitaxel formulation. Potential advantages of polymer-drug conjugates include delivery of a higher concentration of active drug to tumor tissue and limited exposure of normal tissues. In addition, the slow release of drug from the polymer carrier lowers peak plasma concentrations of the active drug. PMID- 17428163 TI - Synthetic retinoid fenretinide in breast cancer chemoprevention. AB - Preclinical models suggest that retinoids inhibit mammary carcinogenesis. The induction of apoptosis is a unique feature of fenretinide, the most-studied retinoid in clinical trials of breast cancer chemoprevention, owing to its selective accumulation in breast tissue and its favorable toxicological profile. In a Phase III breast cancer prevention trial, fenretinide showed a strong trend of reduction of incidence of second breast malignancies in premenopausal women, which was confirmed by 15 years of follow-up. This warrants further research on the mechanisms of action and potential efficacy of fenretinide and provides the rationale for a Phase III primary prevention trial in young women at high risk for breast cancer. This review will highlight the role of fenretinide in breast cancer chemoprevention. PMID- 17428164 TI - Bevacizumab in the management of solid tumors. AB - Angiogenesis is defined as the formation of new blood vessels from a pre-existing vascular bed. By supplying nutrients and oxygen and removing waste products in malignant tumors, it is an essential process that regulates cancer growth and dissemination. This process is regulated by both pro- and antiangiogenic compounds. Vascular endothelial growth factor is one of the most important and best-studied proangiogenic factors. Bevacizumab, a monoclonal antibody against vascular endothelial growth factor, has been shown to inhibit angiogenesis and is proving to be of clinical benefit in a variety of tumor types. The strongest evidence comes from studies in advanced colorectal and non-small-cell lung cancer, with growing evidence in breast and epithelial ovarian tumors. The duration and timing of bevacizumab's use is currently the focus of several ongoing clinical trials. PMID- 17428165 TI - Tariquidar (XR9576): a P-glycoprotein drug efflux pump inhibitor. AB - P-glycoprotein actively transports structurally unrelated compounds out of cells, conferring the multidrug resistance phenotype in cancer. Tariquidar is a potent, specific, noncompetitive inhibitor of P-glycoprotein. Tariquidar inhibits the ATPase activity of P-glycoprotein, suggesting that the modulating effect is derived from the inhibition of substrate binding, inhibition of ATP hydrolysis or both. In clinical trials, tariquidar is tolerable and does not have significant pharmacokinetic interaction with chemotherapy. In patients, inhibition of P glycoprotein has been demonstrated for 48 h after a single dose of tariquidar. Studies to assess a possible increase in toxicity of chemotherapy and the impact of P-glycoprotein inhibition on tumor response and patient outcome are ongoing. Tariquidar can be considered an ideal agent for testing the role of P glycoprotein inhibition in cancer. PMID- 17428166 TI - VIth meeting on cancer-induced bone disease. PMID- 17428167 TI - Perspectives of clinical proteomics in gastrointestinal cancer. AB - Every year approximately 2 million Europeans develop a malignancy; the group of gastrointestinal cancers is the single largest group, with approximately 600,000 new cancers annually. Despite recent innovations and developments in the diagnosis and management of these cancers, prognosis remains poor and treatment options limited. In recent years, new technological advances in proteome analysis and its application to patient management have been made and are the subject of ongoing clinical studies. The identification of biomarkers and biomarker patterns has raised hope that noninvasive diagnosis of cancers in their early stages may soon be an option. However, before the results of proteome analysis can be implemented in the management of cancer patients, further validation of these markers and the issues of sensitivity, specificity, reproducibility and accuracy need to be addressed and solved. PMID- 17428168 TI - Is 18F FDG PET-CT cost effective in lung cancer? AB - Medical advances are often technology driven. One of the new technologies used in imaging is the positron emission tomography (PET)-computed tomography (CT) scanner, which combines the advantages of the functional imaging of PET with CT. This enables both functional and anatomical data to be acquired at the same time. However, such technology is expensive and not always widely available. In the assessment of patients suffering from non-small-cell lung cancer the only curative treatment is surgery or radical radiotherapy, but this is only possible if disease is limited to the affected lung and local nodes. Imaging with CT has not been reliable in determining disease load and many patients undergo futile thoracotomies, which are both expensive and also carry a significant morbidity. In this clinical scenario there is evidence that (18)F fluorodeoxyglucose PET-CT is the best discriminator of disease load and, if used in the correct clinical setting, will not only reduce the number of futile operations but also reduce costs. PMID- 17428169 TI - Chemotherapy for colorectal cancer in the metastatic and adjuvant setting: past, present and future. AB - There have been significant advances in the use of chemotherapy in the treatment of colorectal cancer patients over the last 20 years. Initial improvements in treatment were made with increased understanding of the pharmacology of 5 fluorouracil and the discovery of modulators of its activity (e.g., leucovorin). However, in the last few years the discovery of new cytotoxic drugs with efficacy in large bowel cancer (e.g., oxaliplatin and irinotecan) and monoclonal antibodies (e.g., bevacizumab and cetuximab) have significantly improved patient outcome and prognosis. Systemic chemotherapy in the metastatic setting has been shown to prolong survival and improve quality of life. Chemotherapy now also has a clear role as an adjunct to surgery to improve survival in stage III and certain 'high-risk' stage II colorectal cancer patients. The evolution of chemotherapy use, current practice in the metastatic and adjuvant setting and possible future directions are discussed. PMID- 17428170 TI - Molecular markers of chemotherapeutic response and toxicity in colorectal cancer. AB - Outcomes in colorectal cancer have improved over the last 15 years; this is in part due to the optimization of 5-fluorouracil schedules and the introduction of new and effective chemotherapeutic agents, such as irinotecan and oxaliplatin. However, not all patients respond to these agents and a proportion may suffer severe side effects from particular chemotherapy drugs. These observations have resulted in a concerted research effort to identify markers of chemotherapy efficacy and toxicity. Here we review the evidence for using molecular markers to individualize chemotherapy treatment in colorectal cancer. PMID- 17428171 TI - Predicting response to epidermal growth factor receptor-targeted therapy in colorectal cancer. AB - The discovery over 20 years ago by the Nobel Laureate Stanley Cohen of epidermal growth factor and its receptor, followed by the recognition that this receptor is overexpressed in multiple cancer types, has been of phenomenal significance. From these events the 'Holy Grail' of targeted therapy has looked increasingly realistic. Over the last 5 years this work has come of age with the licensing of multiple agents targeting this important mitogenic pathway in multiple tumor types. However, these agents and the technology behind them, while impressive, have resulted in lower clinical response rates than anticipated. In this review we will focus on the epidermal growth factor receptor-targeted therapies in colorectal cancer, why our expectations from these therapies have not yet been fulfilled and how we may predict those cancers that are likely to respond or be resistant to these therapies through a greater appreciation of the intricacy, diversity and dynamism of cellular signaling mechanisms. PMID- 17428172 TI - Intestinal homeostasis and neoplasia studied using conditional transgenesis. AB - Constitutive mouse models of intestinal neoplasia, such as the Apc(min/+) (multiple intestinal neoplasia) mouse have proven valuable tools both for furthering our understanding of tumorigenesis and for the development of therapeutic strategies. However, the in vivo study of a number of genes has been precluded by their absolute requirement during embryonic development. This has led to the development of conditional strategies that allow gene regulation in vivo. This review describes the principal techniques used to achieve conditional transgenesis within the mouse intestine, with a particular focus upon the Cre-Lox and Tet-regulable systems. Further, we discuss how these techniques are being used to dissect the mechanisms governing both normal homeostasis and neoplastic development within the intestine. PMID- 17428173 TI - Medical treatment of pancreatic cancer. AB - Pancreatic carcinoma is a devastating disease with the worst prognosis of all solid tumors; the only cure is surgery. The vast majority of patients are inoperable at the time of diagnosis and require palliative treatment. With a median survival time oscillating around 6 months, indicating an almost complete resistance to conventional cytotoxic and radiation therapy, there is ample room for improvement. Therefore, pancreatic carcinoma has been used to trial many new substances and novel concepts. All aspects of palliative antitumor treatment will be presented in detail and discussed. Finally, some outlooks are given into the future of pancreatic cancer treatment. PMID- 17428174 TI - Molecular predictors of chemotherapy response in non-small-cell lung cancer. AB - Lung cancer is the number one cause of cancer-related mortality. In order to improve the outcome of patients, advances in the understanding of cancer biology and the development of therapeutic modalities that target key proliferation and survival mechanisms are needed. In vitro data have demonstrated that the genes RRM1 and ERCC1 are important components of these mechanisms. Recently, how these genes affect lung cancer therapy has been explored in the clinical setting with the goal of finding customized treatment algorithms to optimize efficacy, improve outcomes and minimize toxicity. PMID- 17428175 TI - Targeting the vascular endothelial growth factor pathway in the treatment of multiple myeloma. AB - Multiple myeloma is a clonal plasma cell malignancy within the bone marrow associated with bone loss, renal disease and immunodeficiency. Despite new insights into the pathogenesis of multiple myeloma and novel targeted therapies, the median survival remains 3-5 years. It is now well established that the intimate relation between the tumor cells and components of the microenvironment plays a key role in multiple myeloma pathogenesis. Specifically, tumor cells impact the bone marrow and thereby cause immune suppression and lytic bone lesions; conversely, components of the bone marrow provide signals that influence the behavior of multiple myeloma cells, including tumor cell growth, survival, migration and drug resistance. Important contributing effectors are tumor cell stroma cell and cell-extracellular matrix contacts, the bone marrow vasculature, and a variety of cytokines and growth factors in the bone marrow milieu. PMID- 17428176 TI - Superficial bladder cancer: part 2. Management. AB - In the second section of a two-part article, the recent literature is reviewed and the management of nonmuscle-invasive transitional cell carcinoma of the bladder is discussed. Particular attention is given to the indications and timing of intravesical chemotherapy and immunotherapy and the differences in efficacy and side-effect profiles of the available agents. The indications and role of second-look transurethral resection are reviewed. Additionally, the role of bacillus Calmette-Guerin in the management of this disease in terms of definitive treatment and maintenance therapy is discussed. We also offer a review of the literature regarding therapies for bacillus Calmette-Guerin-refractory nonmuscle invasive transitional cell carcinoma of the bladder and their current place in practice. PMID- 17428177 TI - Development of histone deacetylase inhibitors for cancer treatment. AB - Histone deacetylase (HDAC) inhibitors are an exciting new addition to the arsenal of cancer therapeutics. The inhibition of HDAC enzymes by HDAC inhibitors shifts the balance between the deacetylation activity of HDAC enzymes and the acetylation activity of histone acetyltransferases, resulting in hyperacetylation of core histones. Exposure of cancer cells to HDAC inhibitors has been associated with a multitude of molecular and biological effects, ranging from transcriptional control, chromatin plasticity, protein-DNA interaction to cellular differentiation, growth arrest and apoptosis. In addition to the antitumor effects seen with HDAC inhibitors alone, these compounds may also potentiate cytotoxic agents or synergize with other targeted anticancer agents. The exact mechanism by which HDAC inhibitors cause cell death is still unclear and the specific roles of individual HDAC enzymes as therapeutic targets has not been established. However, emerging evidence suggests that the effects of HDAC inhibitors on tumor cells may not only depend on the specificity and selectivity of the HDAC inhibitor, but also on the expression patterns of HDAC enzymes in the tumor tissue. In this review, the recent advances in the understanding and clinical development of HDAC inhibitors, as well as their current role in cancer therapy, will be discussed. PMID- 17428179 TI - Reniform in U.S. cotton: when, where, why, and some remedies. AB - The reniform nematode, Rotylenchulus reniformis, is an emerging problem in U.S. cotton. The impact of this nematode and the extent to which it has and will continue to spread across the U.S. cotton belt are controversial. Long-term changes in cotton production and unique biological attributes of R. reniformis are key factors. Expert opinion surveys indicate that R. reniformis has replaced the root-knot nematode (Meloidogyne incognita) as the major nematode of cotton in Mississippi, Louisiana, and Alabama. In neighboring states the incidence of heavily infested fields has increased during the past 10 years. Estimated annual loss to the U.S. cotton crop is $ 130 M. Crop rotation and nematicides can reduce losses. Introgression of genetic resistance from primitive accessions of other cotton species offers the most promising opportunity to effectively control this pathogen in the long term. Laboratories in several institutions are currently pursuing this goal, with the promise of resistant cultivars adapted to U.S. cotton production regions within three years. PMID- 17428178 TI - The micromachinery of mechanotransduction in hair cells. AB - Mechanical stimuli generated by head movements and changes in sound pressure are detected by hair cells with amazing speed and sensitivity. The mechanosensitive organelle, the hair bundle, is a highly elaborated structure of actin-based stereocilia arranged in precise rows of increasing height. Extracellular linkages contribute to its cohesion and convey forces to mechanically gated channels. Channel opening is nearly instantaneous and is followed by a process of sensory adaptation that keeps the channels poised in their most sensitive range. This process is served by motors, scaffolds, and homeostatic mechanisms. The molecular constituents of this process are rapidly being elucidated, especially by the discovery of deafness genes and antibody targets. PMID- 17428180 TI - Conserved and tissue-specific genic and physiologic responses to caloric restriction and altered IGFI signaling in mitotic and postmitotic tissues. AB - Caloric restriction (CR), the consumption of fewer calories without malnutrition, and reduced insulin and/or IGFI receptor signaling delay many age-related physiological changes and extend the lifespan of many model organisms. Here, we present and review microarray and biochemical studies indicating that the potent anticancer effects of CR and disrupted insulin/IGFI receptor signaling evolved as a byproduct of the role of many mitotic tissues as reservoirs of metabolic energy. We argue that the longevity effects of CR are derived from repeated cycles of apoptosis and autophagic cell death in mitotically competent tissues and protein turnover and cellular repair in postmitotic tissues. We review studies showing that CR initiated late in life can rapidly induce many of the benefits of lifelong CR, including its anticancer effects. We also discuss evidence from liver and heart indicating that many benefits of lifelong CR are recapitulated in mitotic and postmitotic tissues when CR is initiated late in life. PMID- 17428181 TI - ChREBP, a transcriptional regulator of glucose and lipid metabolism. AB - Dysregulations in hepatic lipid synthesis are often associated with obesity and type 2 diabetes, and therefore a perfect understanding of the regulation of this metabolic pathway appears essential to identify potential therapeutic targets. Recently, the transcription factor ChREBP (carbohydrate-responsive element binding protein) has emerged as a major mediator of glucose action on lipogenic gene expression and as a key determinant of lipid synthesis in vivo. Indeed, liver-specific inhibition of ChREBP improves hepatic steatosis and insulin resistance in obese ob/ob mice. Since ChREBP cellular localization is a determinant of its functional activity, a better knowledge of the mechanisms involved in regulating its nucleo-cytoplasmic shuttling and/or its post translational activation is crucial in both physiology and physiopathology. Here, we review some of the studies that have begun to elucidate the regulation and function of this key transcription factor in liver. PMID- 17428182 TI - Multidrug-resistant tuberculous meningitis in an HIV-positive patient: a challenging disease. AB - We report two cases of disseminated multidrug-resistant tuberculosis with meningitis in HIV-positive patients, who were both recent emigrants from sub Saharan Africa. Our two cases highlight new challenges in the care of HIV and tuberculosis coinfection including early diagnosis and treatment of multidrug resistant tuberculosis that is spreading. PMID- 17428183 TI - Changing treatment paradigms: hepatitis C virus in HIV-infected patients. AB - The evaluation of hepatic histology and treatment of hepatitis C virus (HCV)/HIV coinfected patients is rapidly changing. HCV has become an important cause of mortality in HIV-infected patients. Consequently, assessment of liver histology in all coinfected patients is particularly important. The evaluation of hepatic histology is shifting from reliance on the liver biopsy toward noninvasive modalities. Additionally, the importance of HCV-associated morbidity and mortality in HIV-infected patients has prompted increasing numbers of these patients to be HCV treatment candidates. Prospective trials in coinfected patients have reported lower sustained virologic responses compared to HCV monoinfected patients. Consequently, the numbers of coinfected nonresponders to pegylated interferon/ribavirin continues to increase. Because none of the presently available treatment modalities for pegylated interferon/ribavirin nonresponders are clearly efficacious, management decisions must be individualized. The options include, HCV-specific medications, maintenance therapy, the use of alternative interferon formulations, and observation. HCV specific agents, particularly protease and polymerase inhibitors, show early promise in HCV-monoinfected individuals. Their use in coinfected patients, however, is likely to be delayed for several years, and they are likely to be used in combination with interferon. Low-dose pegylated interferon, administered in an attempt to slow fibrosis progression (maintenance therapy), is being evaluated in several large prospective trials in monoinfected and coinfected patients. Observation may be best for nonresponders with relatively mild hepatic histology. In summary, HCV treatment will likely follow the example forged by HIV. In the near future, combinations of different drugs will likely be used simultaneously to result in durable viral suppression. PMID- 17428184 TI - Western blot-indeterminate results in Nigerian patients HIV serodiagnosis: the clinical and public health implication. AB - The clinical and public health implication of HIV Western blot (WB) indeterminate results is yet to be appraised in sub-Saharan Africa, including Nigeria. Using HIV Tri Line Test enzyme-linked immunosorbent assay (ELISA), 1286 patients (600 males and 686 females; age range, 5-60 years) with symptoms suggestive of HIV infection were screened. A total of 1020 (79.3%, 95% confidence interval [CI] 76.8-81.5) of the patients comprising of 514 (85.7%) males and 506 (73.8%) females were HIV seropositive and the difference was statistical significantly (chi(2) = 5.72, df = 1, p < 0.05). Western blot analysis of sera from the 1020 HIV-seropositive individuals using the BIO-RAD NEW LAV-BLOT I specifying World Health Organization (WHO) interpretive criteria, confirmed the HIV serostatus of 815 (79.9%, 95% CI, 77.4-82.4) of them with 205 (20.1%, 95% CI, 17.6-22.6) individuals having indeterminate results consisting of either; 1 env +/- gag +/- pol, gag + pol, gag only or pol only. Of these, 102 (19.8%) were males and 103 (20.4%) were females. Patients aged 11-20 years old recorded the highest percentage of indeterminate results (31.7%, 95% CI, 20.2-43.2) while those aged 21-30 years recorded the least (14.2%, 95% CI, 10.6-17.8) and the difference was statistically significant (chi(2) = 15.73, df = 5, p < 0.05). Result confirmed the limitation of Western blot assays in HIV confirmatory serodiagnosis. After obtaining HIV indeterminate Western blot result, clinicians should consider the total profile for the patient, reassess risk factors for HIV infection, perform a HIV retesting at 3-month intervals for 6 months or use an alternate HIV antibody confirmatory assay and running antibody tests for other human retroviruses. PMID- 17428185 TI - Utilization of health care services in hard-to-reach marginalized HIV-infected individuals. AB - To benefit from HIV treatment advances individuals must utilize ambulatory primary care services. Few studies focus on marginalized populations, which tend to have poor health care utilization patterns. This study examined factors associated with health care utilization in hard-to-reach marginalized HIV infected individuals. As part of a multisite initiative evaluating outreach programs that target underserved HIV-infected individuals, 610 participants were interviewed about their HIV disease, health services utilization, substance use, mental health, and case management. Primary outcomes included ambulatory, emergency department, and inpatient visits. Generalized estimating equations were used in logistic regression analyses. On regression analyses ambulatory visits were associated with having insurance (adjusted odds ratio [AOR] = 2.46), mental health medications (AOR = 7.46), and case management (AOR = 4.81). Emergency department visits were associated with having insurance (AOR = 1.74), homelessness (AOR = 2.23), poor health status (AOR = 2.02), length of HIV infection (AOR = 2.02), mental health care (AOR = 1.47), mental health medications (AOR = 1.59), and heavy alcohol intake (AOR = 1.46). Hospitalizations were associated with high school education (AOR = 1.57), having insurance (AOR = 10.45), homelessness (AOR = 2.18), poor health status (AOR = 2.64), length of HIV infection (AOR = 2.03), and mental health medications (AOR = 1.87). In hard-to reach marginalized HIV-infected individuals, having insurance, case management and mental health care were associated with increased ambulatory visits. These findings support HIV multidisciplinary care with marginalized populations. Understanding factors associated with health care utilization is essential for outreach programs to facilitate engagement in HIV care. PMID- 17428186 TI - Testing practices and knowledge of HIV among prenatal care providers in a low seroprevalence state. AB - As the prevalence of heterosexually transmitted HIV increases among women of childbearing age in the United States, so too does the potential for vertical transmission from mother to child. Early maternal diagnosis and appropriate management are critical to minimizing the risk of perinatal infection. We designed a study to evaluate current prenatal care provider testing practices and knowledge of HIV as it relates to pregnancy in a low seroprevalence state. A written questionnaire was mailed to 642 prenatal care providers in Kentucky. Responses were compared to a similar survey conducted in 1998 and to current federal guidelines for HIV management. Nearly all respondents reported to offer HIV testing to all prenatal patients, demonstrating a marked improvement since 1998 (p < 0.001). However, clinicians did not report adequate follow-up when testing is refused and appear to have limited knowledge of the disease as it relates to pregnancy. Only 9.3% of respondents demonstrated proficiency on two knowledge assessment questions. Those with previous experience treating prenatal patients with HIV were more likely to respond correctly (odds ratio [OR] 3.03; 95% confidence interval [CI] 1.08-8.50). Providers with little experience treating patients with HIV may not possess the basic knowledge required to manage the disease during pregnancy. Additional educational interventions are needed in low seroprevalence areas to ensure the appropriate treatment of all HIV-positive pregnant patients and to minimize the risk of preventable perinatal transmission. PMID- 17428187 TI - Predictors of delayed HIV diagnosis in a recently diagnosed cohort. AB - Delayed diagnosis of HIV is associated with a worse prognosis despite highly active antiretroviral therapy. Many persons with HIV infection are diagnosed late in the disease process. We conducted a study of 119 persons recently diagnosed with HIV infection to determine the association of health literacy and other factors with delayed diagnosis. Patients were recruited from four publicly funded facilities in Houston, Texas. Health literacy was measured with the Test of Functional Health Literacy in Adults (TOFHLA). Delayed diagnosis was assessed by CD4 cell count at diagnosis. Sixty-five percent of patients had CD4 cell counts 350 cells/mm(3) or less. Twenty-eight percent had inadequate health literacy, but literacy was not associated with CD4 cell count. Thirty-eight percent were tested because they "felt sick." In multivariable analysis, female gender (p = 0.005), reason tested other than "felt sick" (p < 0.001), and marijuana use (p = 0.004) and other illicit drug use (p = 0.01) were predictors of having a higher CD4 cell count at diagnosis. These results confirm that late diagnosis of HIV is common among users of public health care facilities. Expanded routine testing for HIV infection is needed with attention directed to men and persons who may not recognize that they are at risk for contracting HIV infection. PMID- 17428188 TI - Urine-based asymptomatic urethral gonorrhea and chlamydia screening and sexual risk-taking behavior in men who have sex with men in greater Boston. AB - To assess the prevalence of asymptomatic urethral gonorrhea and chlamydia men who have sex with men (MSM) living in greater Boston, 206 men attending routine medical appointments consented to urine-based chlamydia and gonorrhea screening using urine LCR amplification. Of those screened, 201 patients also completed a seven-question survey to assess sexual risk behaviors associated with urethral sexually transmitted infections. Less than 1% of the asymptomatic patients screened tested positive for urethral chlamydia; none tested positive for urethral gonorrhea. Forty-eight percent reported multiple sexual partners in the 30 days prior to screening, with HIV-infected patients reporting fewer partners and less unprotected insertive anal sex than HIV-uninfected patients. Almost 25% of patients screened used the Internet in the 30 days prior to screening to find a sexual partner. Internet use was associated with increased numbers of sexual partners in the 30 days prior to screening. Findings suggest that asymptomatic urethral chlamydia and gonorrhea may be uncommon in MSM living in the greater Boston area and that the recent rise in the prevalence of sexually transmitted infections may not be due to untreated asymptomatic infections. Increased awareness of STD symptoms among patients and medical providers is critical to timely diagnosis and treatment of STDs in MSM. PMID- 17428189 TI - Improvement of psychosocial adjustment to HIV-1 infection through a cognitive behavioral oriented group psychotherapy program: a pilot study. AB - The present study evaluated the efficacy of a group therapy program in improving psychosocial adjustment to HIV infection, and tried to identify variables predictive of greater improvement. The outcome of 47 completing patients was analyzed, comparing the measures between T1 (1 month before therapy), and T2 (first session), and between T2 and T3 (last session) using the Wilcoxon matched pairs signed-ranks test for each dimension of the Psychosocial Adjustment to Illness Scale (PAIS). The therapy consisted of 16 weekly 2-hour sessions following a structured time-limited cognitive-behavioral group psychotherapy program. During the intervention (between T2 and T3) a significant improvement was observed in health care orientation, vocational environment, domestic environment, sexual relation, extended family relationships, social environment, and total PAIS. There were no changes during baseline (between T1 and T2) in any of the PAIS subscales, or in the total PAIS score. Sexual route of transmission was independently associated with an improvement in health care orientation (beta = 2.525). Time since HIV diagnosis (beta = 0.022) and being employed (beta = 2.548) were independently associated with an improvement in adjustment to vocational environment. Men who have sex with men showed a poorer improvement in adjusting to family relations after the intervention (beta = -2.548). Finally, a lower CD4 count (beta = -0.005) and being employed (beta = 3.054) were independently associated with an improvement in adjustment to social environment. Our psychotherapy program improved psychosocial functioning in a heterogeneous sample of HIV-1-infected patients referred to a consultation-liaison psychiatry unit. PMID- 17428190 TI - Prezista gets EU approval. PMID- 17428192 TI - Low-intensity ultrasound inhibits apoptosis and enhances viability of human mesenchymal stem cells in three-dimensional alginate culture during chondrogenic differentiation. AB - Many studies have investigated optimal chondrogenic conditions, but only a few of them have addressed their effects on cell viability or the methods to enhance it. This study investigated the effect of low-intensity ultrasound (LIUS), a well known chondrogenic inducer, on the viability of human mesenchymal stem cells (hMSCs) during chondrogenic differentiation in three-dimensional (3-D) alginate culture. The hMSCs/alginate layer was cultured in a chondrogenic defined medium and treated with transforming growth factor-beta1 (TGF-beta1) and/or LIUS for 2 weeks. Along with chondrogenic differentiation for 2 weeks, the 3-D alginate culture and TGF-beta1 treatment resulted in the decrease of cell viability, which appeared to be mediated by apoptosis. In contrast, co-treatment with LIUS clearly enhanced cell viability and inhibited apoptosis under the same conditions. The effect of LIUS on the apoptotic event was further demonstrated by changes in the expression of apoptosis/viability related genes of p53, bax, bcl-2, and PCNA. These results suggest that the LIUS treatment could be a valuable tool in cartilage tissue engineering using MSCs as it enhances cell viability and directs the chondrogenic differentiation process, its well-known activity. PMID- 17428193 TI - The mammalian oxysterol-binding protein-related proteins (ORPs) bind 25 hydroxycholesterol in an evolutionarily conserved pocket. AB - OSBP (oxysterol-binding protein) homologues, ORPs (OSBP-related proteins), constitute a 12-member family in mammals. We employed an in vitro [3H]25OH (25 hydroxycholesterol)-binding assay with purified recombinant proteins as well as live cell photo-cross-linking with [3H]photo-25OH and [3H]photoCH (photo cholesterol), to investigate sterol binding by the mammalian ORPs. ORP1 and ORP2 [a short ORP consisting of an ORD (OSBP-related ligand-binding domain) only] were in vitro shown to bind 25OH. GST (glutathione S-transferase) fusions of the ORP1L [long variant with an N-terminal extension that carries ankyrin repeats and a PH domain (pleckstrin homology domain)] and ORP1S (short variant consisting of an ORD only) variants bound 25OH with similar affinity (ORP1L, K(d)=9.7x10(-8) M; ORP1S, K(d)=8.4 x10(-8) M), while the affinity of GST-ORP2 for 25OH was lower (K(d)=3.9x10(-6) M). Molecular modelling suggested that ORP2 has a sterol-binding pocket similar to that of Saccharomyces cerevisiae Osh4p. This was confirmed by site-directed mutagenesis of residues in proximity of the bound sterol in the structural model. Substitution of Ile249 by tryptophan or Lys150 by alanine markedly inhibited 25OH binding by ORP2. In agreement with the in vitro data, ORP1L, ORP1S, and ORP2 were cross-linked with photo-25OH in live COS7 cells. Furthermore, in experiments with either truncated cDNAs encoding the OSBP-related ligand-binding domains of the ORPs or the full-length proteins, photo-25OH was bound to OSBP, ORP3, ORP4, ORP5, ORP6, ORP7, ORP8, ORP10 and ORP11. In addition, the ORP1L variant and ORP3, ORP5, and ORP8 were cross-linked with photoCH. The present study identifies ORP1 and ORP2 as OSBPs and suggests that most of the mammalian ORPs are able to bind sterols. PMID- 17428194 TI - A highly efficient method for the generation of a recombinant Bombyx mori nuclear polyhedrosis-virus Bacmid and large-scale expression of foreign proteins in silkworm (B. mori) larvae. AB - In the post-genomic era, one of the challenges and a source of competition is the development of high-throughput, large-scale and low-cost eukaryotic cDNA cloning and expression systems. The baculovirus expression system is the most popular one and plays an important role in the high-level expression of eukaryotic proteins. In the present study, a convenient, rapid and highly efficient method for the construction of recombinant BmNPV (Bombyx mori nuclear polyhedrosis virus)-Bacmid vector (BmBacmid) for low-cost protein expression in silkworm (B. mori) larvae was established by using the MAGIC (mating-assisted genetically integrated cloning) strategy. By simply mixing the donor bacteria strain containing the constructed donor vector pCTdual harbouring foreign genes and the recipient strain containing modified BmBacmid, 99.8% positive recombinant BmNPV-Bacmids were obtained. Reporter genes egfp (enhanced green fluorescent protein gene) and DsRed (Discosoma sp. red fluorescent protein gene) and target gene man (beta mannanase gene) encoding beta-mannanase were expressed in the silkworm larvae of B. mori at high level by injection of recombinant BmBacmid DNA directly with the standard calcium phosphate transfection procedure. The possibility of constructing a high-quality baculovirus cDNA library by transferring an ordinal plasmid cDNA library into the recipient BmBacmid in Escherichia coli was explored. PMID- 17428195 TI - Construction of a restriction-endonuclease-Eam1105I-generated T-vector for high throughput cloning and expression. AB - A novel T-vector was constructed that could be used for direct cloning and expression of PCR-amplified cDNA. The technique was based on the insertion into the parent vector of two endonuclease-Eam1105I restriction sequences spaced by an expression cassette of the full-length beta-galactosidase, which helped to improve cloning efficiency and to minimize the non-recombinant background of the T-vector when used to clone PCR products. Moreover, this method took advantage of the reconstitution of the rarest restriction sequence of MssI to enable directional cloning. These advantages make the T-vector suitable for high throughput expression and analysis. PMID- 17428196 TI - Normal levels of Rac1 are important for dendritic but not axonal development in hippocampal neurons. AB - BACKGROUND INFORMATION: Rho family GTPases are required for cytoskeletal reorganization and are considered important for the maturation of neurons. Among these proteins, Rac1 is known to play a crucial role in the regulation of actin dynamics, and a number of studies indicate the involvement of this protein in different steps of vertebrate neuronal maturation. There are two distinct Rac proteins expressed in neurons, namely the ubiquitous Rac1 and the neuron-specific Rac3. The specific functions of each of these GTPases during early neuronal development are largely unknown. RESULTS: The combination of the knockout of Rac3 with Rac1 down-regulation by siRNA (small interfering RNA) has been used to show that down-regulation of Rac1 affects dendritic development in mouse hippocampal neurons, without affecting axons. F-actin levels are strongly decreased in neuronal growth cones following down-regulation of Rac1, and time-lapse analysis indicated that the reduction of Rac1 levels decreases growth-cone dynamics. CONCLUSIONS: These results show that normal levels of endogenous Rac1 activity are critical for early dendritic development, whereas dendritic outgrowth is not affected in hippocampal neurons from Rac3-null mice. On the other hand, early axonal development appears normal after Rac1 down-regulation. Our findings also suggest that the initial establishment of neuronal polarity is not affected by Rac1 down-regulation. PMID- 17428197 TI - Scythe regulates apoptosis through modulating ubiquitin-mediated proteolysis of the Xenopus elongation factor XEF1AO. AB - Scythe was originally identified as a novel Reaper-binding anti-apoptotic protein, although the mechanisms of its functions remain largely obscure. Our previous analysis revealed that Scythe can bind to a proteasomal subunit via N terminal domains and that the domains are required for appropriate development of Xenopus embryos. In the present study, we show evidence that the N-terminus of Scythe interacts with XEF1AO, a maternal form of Xenopus laevis EF1A that was suggested to be a potential inducer of apoptosis in vertebrates, and that the binding enhances the poly-ubiquitin modification and subsequent degradation of XEF1AO. Scythe is required for degradation of XEF1AO, since immunodepletion of Scythe from embryonic extracts stabilized XEF1AO significantly. Furthermore, we show that apoptosis induced by accumulation of XEF1AO can be suppressed by co expression of the full-length form of Scythe. These observations indicate that the proteolytic regulation of XEF1AO, mediated through Scythe, is essential to prevent inappropriate accumulation of XEF1AO and resulting apoptotic events during the course of Xenopus development. PMID- 17428198 TI - Polyamine analogues: potent inducers of nucleosomal array oligomerization and inhibitors of yeast cell growth. AB - Polyamines are naturally occurring intracellular polycations that are essential for viability and growth of eukaryotes. Dysregulation of polyamine metabolism is a hallmark of cancer and the carcinogenic process, and consequently development of polyamine analogues has emerged as a viable strategy for therapeutic intervention. Previously, we showed that the naturally occurring polyamines spermidine and spermine were quite effective at inducing the oligomerization of nucleosomal arrays in vitro, suggesting that polyamines may play a key role in regulating higher order chromatin structures in vivo. Here, we analyse the ability of a number of synthetic polyamine analogues to potentiate formation of higher order chromatin structures in vitro. We find that a class of long-chain polyamines called oligoamines are potent inducers of nucleosomal array oligomerization in vitro and that these same polyamine analogues rapidly block yeast cell growth. PMID- 17428199 TI - Clinical and pathological features of fat embolism with acute respiratory distress syndrome. AB - FES (fat embolism syndrome) is a clinical problem, and, although ARDS (acute respiratory distress syndrome) has been considered as a serious complication of FES, the pathogenesis of ARDS associated with FES remains unclear. In the present study, we investigated the clinical manifestations, and biochemical and pathophysiological changes, in subjects associated with FES and ARDS, to elucidate the possible mechanisms involved in this disorder. A total of eight patients with FES were studied, and arterial blood pH, PaO(2) (arterial partial pressure of O(2)), PaCO(2) (arterial partial pressure of CO(2)), biochemical and pathophysiological data were obtained. These subjects suffered from crash injuries and developed FES associated with ARDS, and each died within 2 h after admission. In the subjects, chest radiography revealed that the lungs were clear on admission, and pulmonary infiltration was observed within 2 h of admission. Arterial blood pH and PaO(2) declined, whereas PaCO(2) increased. Plasma PLA(2) (phospholipase A(2)), nitrate/nitrite, methylguanidine, TNF-alpha (tumour necrosis factor-alpha), IL-1beta (interleukin-1beta) and IL-10 (interleukin-10) were significantly elevated. Pathological examinations revealed alveolar oedema and haemorrhage with multiple fat droplet depositions and fibrin thrombi. Fat droplets were also found in the arterioles and/or capillaries in the lung, kidney and brain. Immunohistochemical staining identified iNOS (inducible nitric oxide synthase) in alveolar macrophages. In conclusion, our clinical analysis suggests that PLA(2), NO, free radicals and pro-inflammatory cytokines are involved in the pathogenesis of ARDS associated with FES. The major source of NO is the alveolar macrophages. PMID- 17428200 TI - The history of anatomy in Persia. AB - The study of human anatomy can be found throughout the rich history of Persia. For thousands of years, morphological descriptions derived from this part of the world have contributed to and have helped form our current anatomical knowledge base. In this article we review the major influential Persian periods and the individuals who have contributed to the development of anatomy. We have divided the history of Persia into five eras: (1) the period of the Elamites, Medes, early Persians and Babylonians (10th millennium to 6th century BC); (2) following the establishment of the Persian Empire (6th century BC) to the 7th century AD; (3) after the Islamic conquest of Persia to the ascendency of Baghdad (7th to 13th century AD); (4) from the Mongol invasion of Persia to the foundations of modern anatomy (13th to 18th century AD); and (5) modern Persia/Iran (18th century AD to present). Evidence indicates that human dissection was commonplace in the first era, which led to a disciplined practice of surgery in the centuries leading to the foundation of the Persian Empire. By the emergence of Zoroastrianism in the Persian Empire, the microcosm theory was widely used to understand internal anatomy in relation to the external universe. The world's first cosmopolitan university and hospital were built in Gondishapur, south western Persia, in the third century AD. Greek and Syriac knowledge influenced the second era. With the gradual ruin of Gondishapur and the foundation of Baghdad following the Islamic conquest of Persia (637-651 AD), a great movement took place, which led to the flourishing of the so-called Middle Age or Islamic Golden Age. Of the influential anatomists of this period, Mesue (777-857 AD), Tabbari (838-870 AD), Rhazes (865-925 AD), Joveini (?-983 AD), Ali ibn Abbas (930 994 AD), Avicenna (980-1037 AD) and Jorjani (1042-1137 AD) all hailed from Persia. There is evidence in the Persian literature as to the direct involvement of these scholars in human dissection. Syro-Indian, Byzantine, Greek, Chinese and Arabic knowledge all influenced the third era. In the fourth period, the first colour illustrated anatomical text (by Mansur, 14th century AD) was compiled. Chinese and Indian anatomical styles were embraced, though there was a strong religious siege of anatomy late in this era. By the 19th century, Persia had entered a new era of modernizing movements and academic contact with the West through the reforms of Mirza Tagi Khan Amir Kabir. Knowledge of anatomy for this region in the 20th century was greatly influenced by Europe and America. PMID- 17428201 TI - Effects of aneuploidy on skull growth in a mouse model of Down syndrome. AB - Adult craniofacial morphology results from complex interactions among genetic, epigenetic and environmental factors. Trisomy causes perturbations in the genetic programmes that control development and these are reflected in morphology that can either ameliorate or worsen with time and growth. Many of the specific changes that occur in Down syndrome can be studied in the Ts65Dn trisomic mouse, which shows direct parallels with specific aspects of adult craniofacial dysmorphology associated with trisomy 21. This study investigates patterns of craniofacial growth in Ts65Dn mice and their euploid littermates to assess how the adult dysmorphology develops. Three-dimensional coordinate data were collected from microcomputed tomography scans of the face, cranial base, palate and mandible of newborn (P0) and adult trisomic and euploid mice. Growth patterns were analysed using Euclidean distance matrix analysis. P0 trisomic mice show significant differences in craniofacial shape. Growth is reduced along the rostro caudal axis of the Ts65Dn face and palate relative to euploid littermates and Ts65Dn mandibles demonstrate reduced growth local to the mandibular processes. Thus, the features of Down syndrome that are reflected in the mature Ts65Dn skull are established early in development and growth does not appear to ameliorate them. Differences in growth may in fact contribute to many of the morphological differences that are evident at birth in trisomic mice and humans. PMID- 17428202 TI - Developmental connections between cranial components and the emergence of the first permanent molar in humans. AB - The age of emergence of the first molar (M1) is a developmental event correlated with many variables of primate life history, such as adult brain size. The evolution of human life history is characterized by the inclusion of childhood, which takes place between weaning and M1 emergence. Children still depend on adults for nutrition due to their small digestive system and their immature brains. By contrast, juveniles are not dependent because of M1 emergence, which enables shifting to adult type diet, and attainment of nearly adult brain size. In this study, developmental connections between M1 emergence and growth of cranial components were explored in two ways in order to understand the developmental basis of their evolutionary connections: (1) differences in growth trajectories of cranial components with respect to M1 emergence and (2) differences between individuals with and without fully emerged M1. Growth of anteroneural, midneural, posteroneural, otic, optic, respiratory, masticatory and alveolar cranial components was analysed in human skulls of individuals aged 0-20 years and in an adult reference skull. Volumetric indices were calculated to estimate size. Two subsamples were selected in order to focus on the transition between deciduous and permanent dentition: those with full deciduous dentition and before M1 reaches the occlusal plane; and those who present M1 in full emergence and no other cheek-tooth at the occlusal plane. The principal results were as follows. (1) Trajectories fitted using the whole sample are characterized by an inflection point that takes place before M1 emergence for neural components and around M1 emergence for facial components. (2) Associations between growth and age tend to be strong in those with full deciduous dentition, and weak in those who present M1 in full emergence. (3) Individuals who present M1 in full emergence are larger than those with full deciduous dentition. (4) Growth of components linked to the central nervous system is not linear until M1 emergence. Individuals who present M1 in full emergence are only larger than individuals with full deciduous dentition by 4-5% of adult size. (5) The alveolar component does not show increments between full deciduous dentition and M1 emergence. (6) When volumetric indices were standardized by age, the growth trajectories of individuals with full deciduous dentition and of those with M1 were not decoupled. In general terms, M1 emergence does not show a strong association with growth of the components that may explain differences in life histories. However, the main changes in neural and alveolar components occur in the first 3 years of life, which may be developmentally connected with M1 crown formation. PMID- 17428203 TI - Ultrastructural and immunocytochemical analyses of osteopontin in reactionary and reparative dentine formed after extrusion of upper rat incisors. AB - Reactionary dentine and reparative dentine are two strategies used by the dentine pulp complex to respond to injury. The reactionary dentine is secreted by original odontoblasts, while the reparative dentine is formed by odontoblast-like cells. Osteopontin (OPN) is a non-collagenous protein usually present in the repair of mineralized tissues. It is likely to be present in newly formed dentine but there are no studies attempting to detect it in reactionary and reparative dentine. The aim of the present study was to examine the ultrastructural characteristics, as well as the presence and distribution of OPN in reactionary and reparative dentine by provoking extrusion of the rat incisor. The right upper incisors of 3-month-old male rats were extruded 3 mm and then repositioned into their original sockets. At 3, 7, 10, 15, 20, 30 and 60 days after surgery, the incisors were fixed in glutaraldehyde-formaldehyde and then processed for scanning and transmission electron microscopy and for immunocytochemistry for OPN. After extrusive trauma, the dentine-pulp interface showed the presence of reactionary and reparative dentine, which varied in aspect, thickness and related cells. OPN was not detected in the physiological and reactionary dentine, while it was strongly immunoreactive in the matrix that surrounded the entrapped cells of reparative dentine. In addition, original odontoblasts subjacent to the physiological dentine contained OPN in their Golgi region. The present findings showed that reparative dentine shares some structural characteristics with primary bone, especially in relation to its OPN content. The odontoblast-like cells resemble osteoblasts rather than odontoblasts. PMID- 17428204 TI - Localization of the spinal nucleus of accessory nerve in rat: a horseradish peroxidase study. AB - The spinal nucleus of the accessory nerve (SNA) comprises the group of somata (perikarya) of motor neurons that supply the sternocleidomastoid and trapezius muscles. There are many conflicting views regarding the longitudinal extent and topography of the SNA, even in the same species, and these disagreements prompted the present investigation. Thirty Sprague-Dawley rats (15 males, 15 females) were used. The SNA was localized by retrograde axonal transport of horseradish peroxidase. Longitudinally, the SNA was found to be located in the caudal part (caudal 0.9-1.2 mm) of the medulla oblongata, the whole lengths of cervical spinal cord segments C1, C2, C3, C4, C5 and rostral fourth of C6. In the caudal part of the medulla oblongata, the SNA was represented by a group of perikarya of motor neurons lying immediately ventrolateral to the pyramidal fibres that were passing dorsolaterally after their decussation. In the spinal cord, the motor neuronal somata of the SNA were located in the dorsomedial and central columns at C1, in the dorsomedial, central and ventrolateral columns at C2 and in the ventrolateral column only at C3, C4, C5 and rostral quarter of C6. The perikarya of motor neurons supplying the sternocleidomastoid were located in the caudal part (caudal 0.9-1.2 mm) of the medulla oblongata ventrolateral to the pyramidal fibres that were passing dorsolaterally after their decussation. They were also located in the dorsomedial and central columns at C1, in the dorsomedial, central and ventrolateral columns at C2 and only in the ventrolateral column at the rostral three-quarters of C3. The perikarya of motor neurons supplying the trapezius muscle were located in the ventrolateral column only in the caudal three-quarters of C2, the whole lengths of C3, C4 and C5, and in the rostral quarter of C6. PMID- 17428205 TI - Microfibrils, elastin fibres and collagen fibres in the human intervertebral disc and bovine tail disc. AB - The distribution of microfibrils was studied immunohistochemically in intervertebral discs taken from young normal human surgical cases and from the bovine tail. Co-localization of microfibrils and elastin fibres was examined by dual immunostaining of fibrillin-1 and elastin. Collagen fibre network orientation was studied by using polarized filters. A similar microfibrillar network was seen in both bovine and human discs with network organization being completely different from region to region. In the outer annulus fibrosus (OAF), abundant microfibrils organized in bundles were mainly distributed in the interterritorial matrix. In addition, the microfibril bundles were orientated parallel to each other and co-localized highly with elastin fibres. Within each lamella, co-localized microfibrils and elastin fibres were aligned in the same direction as the collagen fibres. In the interlamellar space, a dense co localized network, staining for both microfibrils and elastin fibres, was apparent; immunostaining for both molecules was noticeably stronger than within lamellae. In the inner annulus fibrosus, the microfibrils were predominantly visible as a filamentous mesh network, both in the interterritorial matrix and also around the cells. The microfibrils in this region co-localized with elastin fibres far less than in the OAF. In nucleus pulposus, filamentous microfibrils were organized mainly around the cells where elastin fibres were hardly detected. By contrast, sparse elastin fibres, with a few of microfibrils, were visible in the interterritorial matrix. The results of this study suggest the microfibrillar network of the annulus may play a mechanical role while that around the cells of the nucleus may be more involved in regulating cell function. PMID- 17428206 TI - Functional specialisation of the thoracic limb of the hare (Lepus europeus). AB - We provide quantitative anatomical data on the muscle-tendon architecture of the hare thoracic limb (specifically muscle mass, fascicle length, pennation angle, tendon mass and length). In addition, moment arms of major thoracic limb muscles were measured. Maximum isometric force and power of muscles, the moment of force about a joint, and tendon stress and strain were estimated. Data are compared with those from other cursorial mammals. The thoracic limb of the hare consists predominantly of extrinsic musculature with long parallel fascicles, specialised for generating force over a large range. A large shoulder flexor/elbow extensor muscle mass is present, in particular Triceps brachii. The pennate nature of the long head of this muscle suggests it has an important role in stabilising the elbow joint during stance, whilst moment arm curves suggest that it may also play a role in initiating shoulder flexion. In addition, Supraspinatus and Infraspinatus are capable of generating high forces, potentially to stabilise the shoulder joint during the stance phase of locomotion. Supraspinatus may in addition play an important role in forelimb protraction. The Subscapularis muscle was capable of generating surprisingly high forces, suggesting that the hare must be able to withstand/produce high forces during activities that need medio lateral stability, such as turning. Distally, tendons were relatively short, showing little potential for elastic energy storage when compared with both their pelvic limb counterparts and their equivalents in the horse thoracic limb. Thus, a 'stiffer' thoracic limb may be beneficial in terms of behaving like a strut, simply supporting and deflecting the body during high-speed running. This more distal/less proximal distribution of limb mass is also likely to be important in retaining the manipulative/adaptive/non-locomotor capabilities of the limb. PMID- 17428207 TI - The concept of health and health promotion. PMID- 17428208 TI - Stress in children: how fifth-year pupils experience stress in everyday life. AB - In many cases, stress in children takes the form of psychosomatic symptoms. The prevalence of headache in children has increased, as has stomach pain and self reported psychological problems. Many children also experience stress without exhibiting any symptoms, as children perceive, understand and evaluate stress in a variety of different ways. The aim of this study was to describe how fifth-year pupils experience stress in everyday life. An explorative and descriptive design based on phenomenology was chosen to investigate children's lifeworld and lived experience of the phenomenon of stress. The data analysis was based on Giorgi's method. The study comprised 29 children, who were interviewed regarding their experience of stress in everyday life. The results revealed five key constituents: 'fear of being late', 'not having sufficient time', 'physical and mental consequences', 'both a negative and a positive feeling' and 'experiencing significant others' stress'. Together, these key constituents form the overall result. This study demonstrates that stress is a part of children's everyday life. The children described stress that was often acute and linked to situations that occur in everyday life. The results can assist in identifying the factors behind the stress experienced by children. This in turn will facilitate observation and intervention when a child exhibits stress symptoms. PMID- 17428209 TI - Stress in women's daily life before and after a myocardial infarction: a qualitative analysis. AB - BACKGROUND: Little is known about women's perceptions of their daily life before and after a myocardial infarction (MI), especially with regard to stress, which is a risk factor for coronary heart disease (CHD). AIM: To describe and explore women's perceptions of stress before and after an MI. METHOD: Two interviews with women who suffered an MI, the first at the hospital (n = 20) and the second 4-10 months after the MI (n = 14), were analysed using a phenomenographic approach. FINDINGS: The stress emanated either from within themselves (personal traits) or as an effect of their immediate surroundings. The period before the MI was stressful due to the different roles they had to maintain in their private and professional lives. They lost control over their daily life. After hospital discharge they both wanted and needed support, as they were terrified when they returned home. They did not know how much they could do and neither did their relatives, while little or no support was provided by the healthcare professionals. CONCLUSIONS: To prevent CHD in daily life and avoid reinforcing stress, it is important to place greater emphasis on stress as an important risk factor. An understanding of this phenomenon can assist primary healthcare, coronary care unit (CCU) and rehabilitation nurses in supporting these women as well as their partners to adapt their daily lives both before and after an MI. It is essential to formulate and implement individualised treatment plans and to provide support groups for women. PMID- 17428210 TI - Gender differences in predictors of survival in elderly nursing-home residents: a 3-year follow up. AB - This study focus on predicting factors of survival possible to modify by nursing care, and the incidence and mortality rate of nursing-home-acquired pneumonia, allocated to 1, 2 and 3 years of follow ups. The residents consisted of 156 women and 78 men living in special housing for the elderly. Data on chronic disease and medication were obtained at baseline, and activities of daily living (ADL) status, nutritional status and body temperature were assessed. The incidence of pneumonia was noted prospectively for 1 year and retrospectively for the following 2 years. Predictive factors for survival were explored by Cox hazard regression analysis. The results showed that age, functional and cognitive impairment were predictors of mortality irrespective of gender, while poor nutritional status in women and chronic obstructive pulmonary disease, heart disease and medication with sedatives in men were gender-specific predictors. ADL correlated positively with dementia and negatively with S-albumin irrespective of gender, while malnutrition correlated positively with ADL in women and positively with chronic obstructive pulmonary disease in men. To promote the quality of daily living in elderly individuals, it is of importance to improve the capabilities in daily functions and nutritional status, especially in women with functional impairment, and to prevent anxiety particularly in men. The findings also clarify that pneumonia is as common as cerebral vascular insult and heart failure as cause of death in this population. PMID- 17428211 TI - End-of-life care in intensive care units: family routines and environmental factors. AB - The purpose of this study was to describe family care routines and to explore environmental factors when patients die in Swedish intensive care units (ICUs). The main research questions were: what are the physical environmental circumstances and facilities when caring for patients in end-of-life and are there any routines or guidelines when caring for dying patients and their families? A questionnaire was sent to 79 eligible Swedish ICUs in December 2003, addressed to the unit managers. The response rate was 94% (n = 74 units). The findings show that, despite recommendations highlighting the importance of privacy for dying ICU patients and their families, only 11% of the respondents stated that patients never died in shared rooms in their ICU. If a patient dies in a shared room, nurses strive to ensure a dignified goodbye by moving the body to an empty room or to one specially designated for this purpose. The majority (76%) of the units had waiting rooms within the ICU. The study also revealed that there is a need for improvements in the follow-up routines for bereaved families. Many units reported (51%) that they often or almost always offer a follow-up visit, although in most cases the bereaved family had to initiate the follow-up by contacting the ICU. Guidelines in the area of end-of-life care were used by 25% of the ICUs. Further research is necessary to acquire a deeper knowledge of the circumstances under which patients die in ICUs and what impact the ICU environment has on bereaved families. PMID- 17428212 TI - Caring for dying patients outside special palliative care settings: experiences from a nursing perspective. AB - The aim of the study was to describe nurses' experiences in caring for gravely ill and dying patients outside special palliative care settings. Tape-recorded qualitative interviews were conducted with a total of nine nurses in primary home care, community care and hospitals. The interviews were analysed according to phenomenological methodology, which resulted in the three common structures: ambition and dedication, everyday encounters, and satisfaction/dissatisfaction. In the 'everyday encounters' structure, the following key constituents emerged: responsibility, cooperation, experience and knowledge, feelings, and time and resources. The results describe the nurses' high ambitions to give dying patients and their relatives high-quality care. Despite this, they experienced greater or lesser degrees of dissatisfaction because of insufficient cooperation, support, time and resources. They experienced satisfaction through contact with patients and relatives, functioning collegial cooperation, and the knowledge, experience and personal growth the care had given them. The results indicate that nurses need the resources such as time, improved methods of communication and cooperation as well as more support in order to give quality palliative care and achieve satisfaction with the outcome. The need for discussion about the conditions for giving palliative care outside the hospices and other special palliative care settings is also elucidated. PMID- 17428213 TI - Caring and uncaring experiences as narrated by persons with long-term pain after a stroke. AB - The aim of this study was to describe how persons with long-term pain after a stroke experience their care. The study is part of a larger research project concerning a group persons suffering from long-term pain after a stroke. Qualitative interviews were performed with 43 persons suffering from central poststroke pain, nociceptive pain or tension-type headache after their stroke incident. Content congruence emerged among the narratives (n = 43), and therefore the most information-rich ones (n = 23) were selected for deeper investigation by means of qualitative content analysis. The results reveal the patients' need for being respected, understood and supported, also for being given adequate time and information. Accessibility and continuity in the professional contacts and with regard to medical and physical treatment was emphasized. The participants' narratives highlight the importance of the professionals having knowledge of pain and pain management. PMID- 17428214 TI - Quality of life in nursing homes: perception of physically frail elderly residents. AB - The purpose of this research was to disclose the characteristics of quality of life as perceived by physically frail but lucid elderly people living in nursing homes to increase the understanding of the phenomenon of quality of life in this setting. Eight elderly residents living in two nursing homes in Iceland were interviewed on two occasions. The interviews were audio-taped and transcribed to generate text for hermeneutic phenomenological analysis. Observation was also undertaken at both nursing homes to enhance the understanding of the residents' narration. The participants' concern in relation to quality of life emerges in the following main themes: (i) Securing the insecure body; (ii) Seeking solace; (iii) Preparing for departure; and (iv) Affirmation of self. The findings indicate that the phenomenon quality of life is manifold and complex, having many dimensions. The most important aspects of quality of life were for the residents to feel secure in the nursing home, have a place of their own where they could be alone with their thoughts, set their affairs in order and be prepared for death. Furthermore, it mattered to be recognized as an individual with his or her roots in their own respective family and doing meaningful things. These aspects of life in a nursing home contribute to living in a meaningful world in which humanity is preserved. It is important in caring for this particular group of residents in the nursing home to know what matters most in relation to their quality of life. PMID- 17428215 TI - Living with chronic obstructive pulmonary disease (COPD): part II. RNs' experience of nursing care for patients with COPD and impaired nutritional status. AB - This study is the second part of a project with the main purpose of obtaining a deeper understanding of the consequences of living with chronic obstructive pulmonary disease (COPD) concerning meal-related situations and an impaired nutritional status. COPD is a slowly progressive lung disease that results in several complications, including malnutrition. Nutritional status is an important part of COPD treatment, and there are criteria recommended for nutritional assessment and interventions among patients with COPD. Despite this, patients with extreme malnutrition and unnoticed weight loss are reported. The aim of the study was to investigate how Registered Nurses (RNs) in primary care describe nutritional assessment practices and interventions in COPD patients with impaired nutritional status. An interview approach using semistructured questions and case vignettes was chosen. The sample included 19 RNs working specifically with COPD patients. Data from interviews were analysed using qualitative content analysis, and nine categories corresponding to the aim were identified. The RNs reported that their assessment of nutritional status was based largely on intuition. Assessment also included detection of the patients' current beliefs and being sensible about information provision - When and How. Interventions were supportive eating interventions, practical and cognitive participation, and making patients aware of the illness trajectory. An overall category that influenced nursing was respecting patients' feelings of shame and guilt about a self-inflicted disease. It seems that RNs use intuition because of a lack of knowledge of systematic methods of nutritional assessment. The findings also indicate that the RNs attempted to build a relationship of trust with the patients rather than provide early information on sensitive topics (e.g. nutritional information). The study reports areas of nursing care for COPD patients that must be improved in the future. PMID- 17428216 TI - The impact of arthritis on daily life with the patient perspective in focus. AB - BACKGROUND: Rheumatoid arthritis (RA) often has a major impact on many areas of an individuals' life. The unpredictability of the prognosis as well as the day-to day symptoms make it impossible to plan ahead. The aim of this study was to identify the domains of concern in the early course of RA. METHODS: Interviews were carried out according to the focus group method. Twenty-three patients with RA, aged between 27 and 67 years and with a disease duration ranging from 1 to 5 years, were included. Four groups were established, the composition of which was based on age, gender and social situation. RESULTS: Arthritis had a major impact on daily life of the patients, especially on the way they performed activities. It also strongly influenced their mood and social life. The participants tried to cope by 'dosing their activities', 'using new strategies' or to 'stretching their limits'. Their sense of helplessness and uncertainty led to both 'anger' and 'depression'. When activities could no longer be maintained, the role positioning in the family also changed. CONCLUSION: The results of this study support the view that more psychosocial support is required to encourage the recently diagnosed RA patient in the new situation of having a chronic disease. PMID- 17428217 TI - Outcomes for patients with chronic low back pain treated using instrumented fusion. AB - The aim of this study was to describe the pain experience at 1-8 years follow-up of patients who were treated with instrumented fusion because of chronic low back pain, and to evaluate the impact of demographic- and disease-specific variables on pain. The sample comprised 101 of 126 Norwegian patients (response rate 80%), aged 25-60 years (mean 46 years) who were treated in a Swedish hospital between 1993 and 2000. The visual analogue scale and Norwegian Pain Questionnaire were used to evaluate pain. Independent t-tests, anova, correlation and multiple linear regression analyses were performed. Sixteen per cent of patients reported no pain, 17% mild pain, 29% moderate pain and 38% strong to excruciating pain following treatment using instrumented fusion. Demographic- and disease-specific variables explained 29% of the variance in back and hip pain intensities, and 19%, 30%, 6% and 23% of the sensory, affective, evaluative and total pain experience, respectively. Patients who needed to use pain-killing reported more pain than those who did not (p < 0.001). Patients suffering from other chronic conditions also reported more affective pain (p < 0.001). In conclusion, at 1-8 years follow up after instrumented fusion most of the patients suffered from moderate pain or less. Further, amount of pain-killing and comorbidity conditions seemed to be highly related to pain in patients treated with instrumented fusion. PMID- 17428218 TI - Cognitive function and pressure ulcers in hip fracture patients. AB - The primary aim of this study was to analyse the documented frequency of pressure ulcers in relation to the patients' cognitive status during the acute care after a hip fracture. The secondary aim was to investigate to what extent the information about pressure ulcers is passed on to the next caregiver. The patient population consisted of 356 patients being admitted to hospital because of a hip fracture. The patients' cognitive function was assessed with the Short Portable Mental Status Questionnaire. The outcome for patients with severe cognitive dysfunction was compared with the outcome for those without severe cognitive dysfunction. The overall incidence of pressure ulcers was 16%. Twenty-three percent of the patients with severe cognitive dysfunction developed a pressure ulcer during hospitalization compared with 14% of patients without severe cognitive dysfunction (p < 0.05). Although patients with severe cognitive dysfunction had an increased risk of developing pressure ulcers, the severity and the localization of their ulcers were comparable to those without severe cognitive dysfunction. Information regarding the presence of the pressure ulcers in the nurses' report to the next caregiver was lacking for 42% of the patients with ulcers regardless of the cognitive function status. In summary, the incidence of pressure ulcers was increased in hip fracture patients with severe cognitive dysfunction. In the acute setting in routine health care, the caregiver needs a validated and feasible instrument for assessing the patient's cognitive function as well as robust criteria for identifying patients at risk for developing pressure ulcers. PMID- 17428219 TI - Prevalence of violence and its relation to caregivers' demographics and emotional reactions: an explorative study of caregivers working in group homes for persons with learning disabilities. AB - The aim of the study was to investigate the prevalence of violence directed towards caregivers working in group homes for persons with learning disabilities, and to examine the relation between violent incidents and caregivers' demographics such as gender, age, years in service, years at the present workplace and education, as well as emotional reactions to violence expressed by the caregivers. A questionnaire was distributed to all caregivers, i.e. Registered Nurses, assistant nurses and nurse's aides, working in group homes for persons with learning disabilities. The results showed that 31% of the caregivers (n = 120) had been exposed to violence during the preceding year with physical violence being the most common type of violence. All categories of caregivers were exposed to violence and emotional reactions were common. Weak relations were found between reported exposure to violence and various demographics among caregivers, such as age and education. Daytime work was the only independent factor in a regression model predicting violence towards the caregivers. Feelings of powerlessness, insufficiency and anger were the most frequently reported emotional reactions elicited by violent situations. PMID- 17428220 TI - Professional roles and research utilization in paediatric care: newly graduated nurses experiences. AB - The aim of the study was to describe and analyse how newly graduated nurses perceive themselves as professionals and their perceptions of barriers to research utilization when starting to work within paediatric care. The nurses were employed in two paediatric university hospitals in Sweden and had been working as staff nurses for 1-3 months. They answered two questionnaires: The Professional Self-Description Form and the Barriers Scale. The results showed that in the assessment of their professional level the highest scores were found in consideration, ambition and sensitivity, and the lowest scores in grasp of ideas, leadership and discrimination. In perceptions of research utilization (Barriers Scale), the items with the most common barriers were 'there is insufficient time on the job to implement new ideas', 'the nurse does not have time to read research' and 'the facilities are inadequate for implementation'. These results indicate that newly graduated nurses seem to be prepared for the nursing profession when it concerns themselves as human individuals but not so prepared for the practice in the real day-to-day work. The most common barriers in the research utilization were connected to the work organization and not to themselves as individuals. The evaluation of professional self, barriers to research utilization and the relationship between these aspects indicates a need for further investigation into the significant meaning of professional self and how this influences the implementation and use of research in everyday work. The results so far also raise the question of how to best support and develop newly graduated nurses in their professional role, to help them to be the highly competent nurses that we need in providing the best and evidence-based care to those who need it. PMID- 17428221 TI - Nursing gaze as framework for nursing practice: a study from acute care settings in Korea, Norway and the USA. AB - The aim of the study was to investigate the nature of nurses' clinical practice in terms of what frames their clinical engagement, and to examine how clinical constructions were made. The study is based on a descriptive design using a clinical field approach in acute care settings in Korea, the USA and Norway. A theoretical sample consisted of between four and six Registered Nurses in each country: up to three from medical wards and three from surgical wards. Data were collected through participant observation of and in-depth interviews with the nurses, as well as from nursing documentation about the patients. The result showed an overall model of a theory of nursing practice consisting of three processes: nursing gaze, clinical construction and clinical engagement. This first article outlines the overall descriptive theory of nursing practice and provides a detailed description of the first aspect: the nursing gaze. The model of the structuring of nursing gaze is set within the philosophy of nursing, consisting of the ontology of client and the ontology of practice. The dimensions of normality and needs constitute the ontology of client, and the dimension of clinical expectations constituted the ontology of practice. PMID- 17428222 TI - Factors influencing satisfaction and well-being among parents of congenital heart disease children: development of a conceptual model based on the literature review. AB - AIM AND METHOD: The treatment and management of congenital heart disease (CHD) has improved dramatically over the past 25 years, necessitating re-evaluation of satisfaction with care and well-being among CHD children and their parents (PCCHD). The present study reviews the published literature over the past 25 years on parental satisfaction with the paediatric care of CHD and well-being among the parents, with the specific aim of: (a) assessing the extent of psychosocial problems and grade of satisfaction with care and (b) modelling factors associated with satisfaction and well-being among the parents. RESULTS: There is general agreement in the literature that PCCHD experience psychosocial morbidity to a higher degree than parents of children with other paediatric conditions and parents of healthy children. The research on satisfaction with care among PCCHD is not conclusive, though there is considerable agreement that a substantial proportion of PCCHD may not be receiving adequate information regarding the ill-child's condition, treatment and medical prognosis. Finally, based on the review of factors affecting satisfaction and well-being, a model is generated indicating that interactions between parental perception of CHD, psychosocial resources and social vulnerability may account for differences in well-being among PCCHD, which in turn may explain differences in satisfaction with care among them. CONCLUSION: A holistic approach to the care of CHD that acknowledges the role of parents' perception of CHD, need for psychosocial resources and social vulnerability in the adaptation process is recommended to improve parental satisfaction with the care of CHD. PMID- 17428223 TI - Parents' postnatal sense of security (PPSS): development of the PPSS instrument. AB - BACKGROUND: There is a need to develop an instrument that measures both the parents' experiences and sense of security during the first postnatal week. No instrument measuring positive dimensions which can be influenced and supported by the postnatal health care has been developed. OBJECTIVE: The aim of the study was to develop a specific instrument to assess both mothers' and fathers' postnatal sense of security concerning the first postnatal week. DESIGN: The study has a methodological and developmental design and was carried out in four steps: construction of the items, face validity, data collection and data analysis. SETTINGS AND PARTICIPANTS: One hundred and thirteen mothers who had given birth in hospitals in southern Sweden and 99 fathers comprised the study participants. DATA ANALYSIS: Statistical analysis, testing for construct validity with explorative factor analysis, internal consistency reliability and comparative validity was carried out. RESULTS: The parents' postnatal sense of security (PPSS) instrument, mother's version, was reduced from 37 to 18 items (explained variance 66.8%, Cronbach's coefficient alpha 0.88) comprising the following dimensions: a sense of the midwives'/nurses' empowering behaviour, a sense of general well-being, a sense of affinity within the family and a sense that breast feeding was manageable. The father's version was reduced from 36 to 13 items (explained variance 69%, Cronbach's coefficient alpha 0.77), and comprised the following dimensions: a sense of the midwives'/nurses' empowering behaviour, a sense of the mother's general well-being including breast feeding, a sense of general well-being and a sense of affinity within the family. CONCLUSIONS: The PPSS instrument is valid and reliable and the only specific instrument measuring postnatal sense of security that is useful for both parents. The instrument needs to be further evaluated. PMID- 17428224 TI - 'Oral health is not my department'. Perceptions of elderly patients' oral health by general medical practitioners in primary health care centres: a qualitative interview study. AB - The purpose of this study was to explore general medical practitioners' (GPs) perceptions of the oral health of their elderly patients. The design was a qualitative study based on individual in-depth interviews with GPs. The criterion for inclusion in the study was that the GP was a specialist in family medicine working in a primary health care centre (PHCC:s) in the county of Stockholm. The participants took part in the study after informed consent. Eleven GPs were interviewed. The interview started with semi-structured questions about the respondents' clinical presentation of their elderly patients', e.g. medication, medical treatment and socioeconomic status. The interview concluded with questions about the respondents' experiences of and perceptions of the oral health of their patients. This process started with the first interview and proceeded with successive interviews until no new relevant information was forthcoming. The initial semi-structured part of the interview guide was analysed for content with special reference to descriptive answers. The final open questions were analysed by a method inspired by grounded theory (GT) and comprised three stages: open coding, axial coding and selective coding. In the GT influenced analysis process, three categories, health perspective, working conditions and cultural differences, each in turn containing subcategories, were identified and labelled. The most significant category, cultural differences, was identified as the core category, explaining the central meaning of the respondents' perceptions of the oral health of their elderly patients. The GPs in this study showed little or no awareness of the oral health of their elderly patients. The interviews disclosed several contributing factors. Barriers to closer integration of oral and general health in the elderly were identified. There existed a cultural gap between the disciplines of dentistry and medicine, which does not enhance and may be detrimental to the overall well-being of elderly patients. PMID- 17428225 TI - Relationships between demographic, clinical and pain variables and health-related quality of life in patients with chronic low back pain treated with instrumented fusion. AB - The purpose of this cross-sectional study was to assess the relationships between demographic, clinical and pain variables and health-related quality of life (HRQOL) in patients who had undergone instrumented spinal fusion because of chronic low back pain (CLBP). The sample comprises 101 patients (70% women, mean age 46 years) who had received surgery between 1993 and 2000. The SF-36 Health Survey was used to assess HRQOL. Pain was assessed by sensory, affective and evaluative subgroups of the Norwegian Pain Questionnaire (modified McGill Pain Questionnaire). Age, work status, number of years after surgery, physical training, other chronic conditions and affective and evaluative pain (all p < 0.05) were the most prominent factors affecting HRQOL. The highest adjusted R(2) to explain the variance was physical function (52%) and the lowest was emotional role limitations (15%). Pain, particularly affective and evaluative pain, contributed more to the explained variance of HRQOL than demographic and clinical variables. PMID- 17428226 TI - A Nordic meeting on G-protein-coupled receptors: from molecular aspects to novel therapy. PMID- 17428227 TI - G-protein-coupled receptors: an update. AB - The receptors that couple to G proteins (GPCR) and which span the cell membranes seven times (7-TM receptors) were the focus of a symposium in Stockholm 2006. The ensemble of GPCR has now been mapped in several animal species. They remain a major focus of interest in drug development, and their diverse physiological and pathophysiological roles are being clarified, i.a. by genetic targeting. Recent developments hint at novel levels of complexity. First, many, if not all, GPCRs are part of multimeric ensembles, and physiology and pharmacology of a given GPCR may be at least partly guided by the partners it was formed together with. Secondly, at least some GPCRs may be constitutively active. Therefore, drugs that are inverse agonists may prove useful. Furthermore, the level of activity may vary in such a profound way between cells and tissues that this could offer new ways of achieving specificity of drug action. Finally, it is becoming increasingly clear that some of these receptors can signal via novel types of pathways, and hence that 'GPCRs' may not always be G-protein-coupled. Thus there are many challenges for the basic scientist and the drug industry. PMID- 17428228 TI - Seven transmembrane receptors: something old, something new. AB - Receptors for hormones, neurotransmitters, drugs, sensory stimuli and many other agents represent the gateway to cellular metabolism and activity. They regulate virtually all physiological processes in mammals. Yet as recently as 40 years ago their very existence was still in question. One class of receptors, those coupled to G proteins (also known as GPCRs or seven transmembrane receptors) comprise by far the largest group (approx. 1000), and are the most important target of clinically used drugs. Here I provide a very personal retrospective of research over the past 35 years which ultimately led to the identification, purification, reconstitution and cloning of the adrenergic receptors; the discovery of their homology with the seven transmembrane spanning visual light receptor rhodopsin and the realization that there was a large gene family of G protein coupled receptors; the elucidation of the molecular mechanisms of receptor desensitization and signalling through G protein-coupled receptor kinases and beta-arrestins; and the appreciation that the structure, signalling, and regulatory mechanisms of the receptors are all highly conserved across the large receptor superfamily. PMID- 17428229 TI - Mining the gene repertoire and ESTs for G protein-coupled receptors with evolutionary perspective. AB - The purpose of this article was to review recent progress in mining the gene repertoire and expressed sequence tags (ESTs) for the super-family of G protein coupled receptors (GPCRs) in the form of a proceeding from the Nordic GPCR meeting held at the Nobel Forum, Karolinska Institute in August 2006. We update and give an overview of the expansion of the main families of GPCRs; Glutamate, Rhodopsin, Adhesion, Frizzled and Secretin (GRAFS) in perspective of fully sequenced genomes. We look into the most recent findings including the work that has been carried out on the spotted green puffer fish (Tetraodon nigroviridis), mouse (Mus musculus), chicken (Gallus gallus), slime mold (Dictyostelium discoideum) and the plant pathogenic fungus Magnaporthe grisea. We use examples from our recent work on chicken GPCRs to highlight the importance of detailed assembly and curation of sequences and how that can affect percentage similarity and phylogeny. ESTs can give valuable information about expression patterns. GPCRs have comparatively low numbers of EST suggesting that GPCRs are in generally expressed in lower amount than other genes. We discuss similarities in the evolution of the trace amine associated receptors with other sensory receptors. PMID- 17428230 TI - Novel aspects of G-protein-coupled receptor signalling--different ways to achieve specificity. AB - Our understanding of signal transduction via G-protein-coupled receptors (GPCR) has developed dramatically during the last decades. The initial idea of linear signalling pathways transferring information from the cell membrane to the nucleus has evolved into a complicated network of signalling pathways offering the possibility of crosstalk, fine tuning and specific regulation at multiple levels. During the recent meeting on GPCRs at the Karolinska Institutet, Stockholm novel aspects of GPCR signalling were presented and discussed. Here, we will discuss several possibilities for GPCRs to achieve specificity in signal transduction, such as the phenomenon of biased agonism, receptor multimerization, the role of co-receptors, the regulation of heterotrimeric G proteins as well as multiple G(s)-dependent pathways to extracellular single-regulated protein kinases. PMID- 17428231 TI - Regulation of G protein-coupled receptor trafficking. AB - G protein-coupled receptors (GPCRs) mediate cellular responses to diverse extracellular stimuli to play a vital role in the control of physiology and behaviour. GPCR trafficking is of fundamental importance for the regulation of GPCRs signaling. In this mini review, we will discuss some of the recent findings on the mechanisms that regulate GPCR trafficking, which include (i) large dense core vesicle (LDCV)-associated GPCR delivery which could be a general cell biological mechanism for rapid modulation of membrane receptors in response to certain stimuli; (ii) lateral diffusion of GPCRs in the plasma membrane for rapid change of the number of neurotransmitter receptors during synaptic plasticity and (iii) constitutive internalization of GPCRs, that contributes to receptor resensitization and distribution, including axonal polarization. PMID- 17428232 TI - Depletion of the lipid raft constituents, sphingomyelin and ganglioside, decreases serotonin binding at human 5-HT7(a) receptors in HeLa cells. AB - AIM: The localization and function of several G protein-coupled receptors, including beta-adrenergic receptors and NK 1 receptors, are regulated via lipid rafts in the plasma membrane. These domains are enriched in cholesterol, gangliosides and sphingolipids, and play an important role in regulating signal transduction in most cell types. Serotonin (5-hydroxytryptamine, 5-HT), acting via 14 different receptors, regulates as diverse effects as mood, metabolism and smooth muscle contraction. 5-HT(7) receptors are involved in the regulation of depression, circadian rhythms, thermoregulation and vasodilatation. Ligand binding and signalling via the 5-HT(7) receptor are regulated by membranous cholesterol. Here we investigated the role of sphingomyelin and gangliosides on binding of 5-HT to 5-HT(7) receptors to further examine the role of lipid raft constituents on 5-HT(7) receptor function. METHODS: HeLa cells stably transfected with the human 5-HT(7) receptor were treated with Fumonisin B(1) or (+/-)-threo-1 Phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP) to reduce sphingomyelin or ganglioside levels, respectively. The effects of these treatments were investigated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) viability assay, cholesterol analysis and [(3)H]5-HT binding studies on intact cells. RESULTS: Treatments with 20 mum Fumonisin B(1) for 24 h or with 10 mum PDMP for 48 h had no effects of total levels if 5-HT(7) receptors, but caused significant decreases in maximum [(3)H]5-HT binding to 5-HT(7) receptors. The effects were cholesterol-independent as levels of cholesterol remained unaffected by either treatment. CONCLUSION: These data demonstrate a role for sphingomyelin and gangliosides in regulating binding of [(3)H]5-HT to 5-HT(7) receptors. These observations further strengthen that actions of 5-HT via 5-HT(7) receptors are dependent upon lipid raft integrity. PMID- 17428233 TI - Inhibition of endocytosis blocks Wnt signalling to beta-catenin by promoting dishevelled degradation. AB - AIM: The Wnt/Frizzled signalling pathway is highly conserved through evolution. Frizzled, the receptors for Wnts, have the topology of seven transmembrane spanning domain receptors. An important means of regulation of these receptors is internalization and desensitization through clathrin-mediated endocytosis. Therefore, we investigated the effects of endocytosis inhibition on Frizzled4 green fluorescent protein (FZD(4)-GFP) localization, dishevelled levels and Wnt 3a signalling to beta-catenin. METHODS: Experiments were performed in the mouse neuronal cell line SN4741 that has previously proven to be valuable for the investigation of Wnt/Frizzled signalling. FZD(4)-GFP distribution has been examined using confocal laser scanning microscopy. Dishevelled protein expression levels and the activation of beta-catenin upon treatment with endocytosis inhibitors (hyperosmolaric sucrose and K(+) depletion), kinase inhibitors and Wnt 3a were analysed by immunoblotting. RESULTS: Hyperosmotic sucrose and K(+) depletion increased the membrane localization of FZD(4)-GFP, and in parallel triggered fast (1-2 h) and almost complete (approx. 95%) degradation of endogenous dishevelled, which was independent of Wnt-induced, CK1-mediated phosphorylation of dishevelled. In addition, dishevelled depletion induced by endocytosis inhibition completely prevented canonical signalling by Wnt-3a to beta-catenin even when osmotic conditions and endocytosis were reverted to normal. CONCLUSIONS: The data provide evidence for a molecular mechanism that could be a basis for a novel negative feedback loop within the Wnt/Frizzled pathway depending on dishevelled degradation. The identification of molecular details of regulatory mechanisms for the Wnt/Frizzled signalling pathway increases our understanding of pathway regulation, which might be of special physiological significance for embryonic development, cancer and neurological disorders. PMID- 17428234 TI - Sex differences in mouse heart rate and body temperature and in their regulation by adenosine A1 receptors. AB - AIM: To examine cardiac function, body temperature and locomotor behaviour in the awake adenosine A(1) receptor knock out mouse of both sexes. METHODS: Male and female A(1)R (+/+) and (-/-) mice, instrumented with telemetric devices, were recorded during basal conditions and after drug administration. RESULTS: Female mice had higher heart rate, body temperature and locomotion, both during daytime and during the night. Awake A(1)R (-/-) mice had a slightly elevated heart rate, and this was more clear-cut in males. Heart rate was also higher in Langendorff perfused denervated A(1)R (-/-) hearts. Body temperature was higher in A(1)R (-/ ) males and females; locomotor activity was higher in A(1)R (-/-) females, but not in males. The adenosine receptor agonist R-PIA (0.2 mg kg(-1)) decreased heart rate and body temperature, but less in A(1)R (-/-) animals than in A(1)R (+/+) mice (P < 0.001 in both parameters). The unselective adenosine receptor antagonist caffeine had a minor stimulatory effect on heart rate in lower doses, but depressed it at a dose of 75 mg kg(-1). Body temperature was increased after a low dose (7.5 mg kg(-1)) of caffeine in both sexes and genotypes, and markedly reduced after a high dose (75 mg kg(-1)) of caffeine. An intermediary dose of caffeine 30 mg kg(-1) increased or decreased body temperature depending on genotype and sex. Locomotor responses to caffeine were variable depending both on genotype and sex. CONCLUSION: Thus, the adenosine A(1) receptor is involved in the regulation of heart rate, body temperature and locomotor activity, but the magnitude of the involvement is different in males and females. PMID- 17428235 TI - Adenosine is not a direct GHSR agonist--artificial cross-talk between GHSR and adenosine receptor pathways. AB - AIM: To assess if adenosine is a direct growth hormone secretagogue receptor (GHSR) agonist by investigating the mechanism behind adenosine induced calcium release in human embryonic kidney 293s (HEK) cells expressing GHSR. METHODS: Calcium mobilization, cyclic adenosine monophosphate (cAMP) and IP(3) experiments were performed using HEK cells stably expressing GHSR and/or adenosine A(2B) receptor (A(2B)R). RESULTS: Adenosine has been widely reported as a GHSR agonist. In our hands, adenosine and forskolin stimulated calcium release from IP(3) controlled stores in HEK-GHSR cells but not in non-transfected HEK cells. This release was not accompanied by increased IP(3) levels. The calcium release was both cholera toxin and U73122 sensitive, indicating the involvement of both Galpha(s)/adenylyl cyclase and Galpha(q/11)/phospholipase C pathways. Importantly, the GHSR inverse agonist [D-Arg(1) D-Phe(5) D-Trp(7,9) Leu(11)] Substance P (SP-analogue) blocked the adenosine stimulated calcium release, demonstrating that GHSR is involved. Assessment of the GHSR-dependent calcium release using adenosine receptor agonists and antagonists resulted in a rank order of potencies resembling the profile of A(2B)R. A(2B)R over-expression in HEK-GHSR cells enhanced potency and efficacy of the adenosine induced calcium release without increasing IP(3) production. Moreover, A(2B)R over-expression in HEK cells potentiated NECA-induced cAMP production. However, GHSR expression had no effect on intracellular cAMP production. CONCLUSION: In HEK-GHSR cells adenosine activates endogenously expressed A(2B)R resulting in calcium mobilization. We hypothesize that the responsible mechanism is cAMP-dependent sensitization of IP(3) receptors for the high basal level of IP(3) caused by GHSR constitutive activity. Altogether, our results demonstrate that adenosine is not a direct GHSR agonist. PMID- 17428236 TI - Eliminating the antilipolytic adenosine A1 receptor does not lead to compensatory changes in the antilipolytic actions of PGE2 and nicotinic acid. AB - AIM: We examined whether compensatory changes after adenosine A(1) receptor knockout [A(1)R (-/-)] eliminate the antilipolytic actions mediated by this receptor. METHODS: Lipolysis experiments were performed on adipocytes prepared from the wild type A(1)R (+/+), A(1)R (-/-) and heterozygous mice. Gene expression was assayed with cDNA microarray technique and real-time PCR; protein expression with immunoblotting. RESULTS: The A(1)R was the only adenosine receptor involved in lipolysis. The effects of adenosine deaminase and 2 chloroadenosine were abolished in A(1)R (-/-) mice. The IC(50) value of 2 chloroadenosine doubled from 16.6 to 33.6 nm when half of the A(1)Rs were eliminated. Adrenergic alpha(2) agonists had no effects on lipolysis. Prostaglandin E(2) (PGE(2)) inhibited lipolysis with an IC(50) value of 5.8 nm (4.7-7.2 nm) in the A(1)R (+/+) mice and 10.6 nm (9.0-12.6 nm) in the A(1)R (-/-) mice. Nicotinic acid inhibited lipolysis with an IC(50) value of 0.30 microm (0.19-0.46 microm) in the A(1)R (+/+) mice and 0.24 microm (0.16-0.37 microm) in the A(1)R (-/-) mice. G(i)alpha(1) mRNA was significantly up-regulated in adipose tissue from A(1)R (-/-) mice. However, immunoblotting showed that G(ialpha1) was not up-regulated at the protein level. CONCLUSION: The A(1)R mediates the antilipolytic actions of adenosine. Deletion of the A(1)R in mice does not result in compensatory increases in G-protein-mediated antilipolytic actions of PGE(2) or nicotinic acid. PMID- 17428237 TI - Solitary plasmacytoma and multiple myeloma: adhesion molecule and chemokine receptor expression patterns. PMID- 17428238 TI - Dual inhibition of ras and bcr-abl signalling pathways in chronic myeloid leukaemia: a phase I/II study in patients in complete haematological remission. AB - Zoledronic acid inhibits the prenylation of ras-related proteins downstream of bcr-abl and preclinical studies have shown augmentation of the inhibitory effects of imatinib in BCR-ABL expressing cells. A Phase I/II study was designed to assess the safety and efficacy of the addition of zoledronic acid to imatinib in patients with chronic myeloid leukaemia (CML) with a suboptimal response to imatinib alone. Ten patients with CML who had been treated with imatinib for at least 2 years and had achieved and maintained a complete haematological response were included. Zoledronic acid was administered intravenously on one occasion every 28 d. The initial dose of 4 mg was given for three consecutive months; in the absence of significant toxicity and/or response the dose was escalated to 8 mg for an additional 3 months. Efficacy was assessed by serial monitoring of blood levels of BCR-ABL transcripts and bone marrow cytogenetics. Addition of zoledronic acid to imatinib caused no haematological toxicity. There were no grade III or IV non-haematological adverse effects. Grade I fatigue, hypocalcaemia and fever were common side effects. No responses were demonstrated after 6 months on the combination. PMID- 17428239 TI - Ultrasound probe positioning to minimize the risk of recto-urethral fistula during cryosurgical ablation of prostate cancer. AB - OBJECTIVE: To evaluate the ability to increase the distance between the rectal mucosa and prostate, and thus decrease the risk of recto-urethral fistula, and to improve the ability to adequately freeze beyond the prostatic capsule during cryosurgical ablation for prostate cancer. PATIENTS AND METHODS: The transrectal ultrasound probe was manipulated to increase the distance from the rectal mucosa to prostate in 28 men scheduled for cryosurgical ablation for localized prostate cancer. Ten patients were treated for local recurrence after previous definitive radiotherapy, and cryosurgery was chosen as the primary treatment for the remainder. RESULTS: The mean (range) distance from the rectal mucosa to the prostate when the probe was securely applied to the rectal wall was 2.5 (1-7) mm, and could be extended to 9.6 (7-14) mm before the ultrasonogram quality diminished enough to impede accurate placing of the cryoprobe. The mean distance gained as a margin of error was 7.1 (4-12) mm (P < 0.001). In no patient was it felt that visualization during the freezing cycles was impaired. No recto urethral fistula was identified. CONCLUSION: The manoeuvres described here allow the surgeon to freeze beyond the prostatic capsule while maintaining a visible distance from rectal wall. The extra distance created by this manoeuvre might reduce the risk of recto-urethral fistula, and potentially improve tumour control in the posterior aspect of the prostate, based on the ability to freeze beyond the capsule to reach lethal temperatures in all prostatic tissue. PMID- 17428240 TI - Evaluation of peri-operative peripheral and renal venous levels of pro- and anti angiogenic factors and their relevance in patients with renal cell carcinoma. AB - OBJECTIVE: To evaluate peri-operative peripheral and renal venous plasma levels of vascular endothelial growth factor (VEGF), platelet-derived growth factor type BB (PDGF-BB), transforming growth factor (TGF)-beta1, endostatin, and thrombospondin-1 (TSP-1) in relation to pathological variables and prognosis, as pro- and anti-angiogenic factors are important for tumour growth and treatment of patients with renal cell carcinoma (RCC). PATIENTS AND METHODS: The study included 74 consecutive patients with sporadic RCC who had tumour nephrectomy. Peripheral venous blood was drawn 1 day before, immediately and 1, 3 and 5 days after surgery. Renal venous blood was collected in a subgroup of 33 patients during surgery. The variables were analysed using quantitative enzyme-linked immunoassay kits, and associated with pathological variables and disease-specific survival. RESULTS: Soon after surgery, peripheral venous VEGF, PDGF-BB and TGF beta1 levels were decreased, whereas endostatin levels were significantly increased. Renal venous VEGF, PDGF-BB and TGF-beta1 levels were higher than in the general venous blood pool. Renal venous VEGF levels were correlated with tumour diameter and associated with grade and vascular invasion. After a mean follow-up of 30 months, higher peripheral preoperative, early peripheral postoperative and renal venous VEGF levels were associated with a poorer prognosis. However, in a multivariate analysis only Tumour-Node-Metastasis stage and Eastern Cooperative Oncology Group performance status were independent prognosticators of disease-specific survival. CONCLUSIONS: Circulating pro- and anti-angiogenic factors change early after nephrectomy. VEGF, PDGF-BB and TGF beta1 are higher in the renal vein than in the general venous blood pool. Higher renal venous and peripheral levels of VEGF might be associated with a poorer prognosis. PMID- 17428241 TI - Saturation biopsy for detecting and characterizing prostate cancer. PMID- 17428242 TI - Molecular characterization of M2 and M3 muscarinic receptor expression in bladder from women with refractory idiopathic detrusor overactivity. AB - OBJECTIVE: To examine the expression of muscarinic M2 and M3 receptors in human bladder detrusor and mucosa, from controls and patients with idiopathic detrusor overactivity (IDO), as antimuscarinic agents are the primary pharmacological treatment for IDO. PATIENTS AND METHODS: Biopsies from the bladder body were collected at cystoscopy from 20 women with urodynamically confirmed refractory IDO (age range 25-86 years); biopsies were also collected from 30 asymptomatic female controls (age range 32-87 years). Samples were collected into RNA extraction medium and dissected into mucosa (urothelium plus lamina propria) and detrusor. RNA was extracted and the expression of M2 and M3 receptor mRNA determined by quantitative competitive reverse transcription-polymerase chain reaction. Results were normalized to beta-actin expression in the same sample. RESULTS: Expression of M3 receptor mRNA, in mucosa of IDO patients (median 0.057 pg M3/100 ng total RNA; interquartile range 0.03-0.13, 12 samples), was four times (P = 0.039, Mann-Whitney) lower than from the control (median 0.22 pg M3/100 ng total RNA; 0.13-0.51, 11 samples). The expression of muscarinic M3 receptor mRNA was higher (14-35 times) in detrusor (control median 3.17; 26 samples) than in mucosa and did not change in IDO (median 2.03; 14 samples). M2 expression was not significantly different with region or with IDO. CONCLUSIONS: These data show that M3 muscarinic receptor mRNA expression was significantly less in mucosa from IDO patients than from age-matched controls. The role of mucosal M3 receptors is unknown at present and elucidation of this role might provide a greater understanding of the aetiology of IDO. PMID- 17428243 TI - Bone resorption and renal calcium reabsorption in renal cell carcinoma-bearing mice: the effects of bisphosphonate. AB - OBJECTIVE: To examine the contribution of the skeleton and the kidney to the development of humoral hypercalcaemia of malignancy (HHM) in a mouse model of HHM treated with a potent bisphosphonate. MATERIALS AND METHODS: Mice bearing the human RCC cell line RC-9 were treated with bisphosphonate (subcutaneous, 0.25 mg/kg body weight olpadronate) or saline solution. Treatment was initiated at a tumour volume (TV) of approximately 100 mm(3) and 500 mm(3), and the mice were monitored for approximately 4 weeks. Serum calcium and phosphate concentrations and trabecular bone volume (TBV) were assessed during and/or after treatment. RESULTS: Athymic mice implanted with the RCC RC-9, developed severe hypercalcaemia and bone resorption. During tumour growth the mean (sd) serum calcium concentration increased to 4.1 (0.3) mmol/L, and phosphate decreased to 1.6 (0.3) mmol/L, vs 2.3 (0.1) and 2.9 (0.4) mmol/L in controls, respectively. TBV decreased from 8.7 (1.8)% in mice with no tumour, to 5.3 (2.7)% in RC-9 bearing mice. Olpadronate initiated at a Tv of 100 mm(3) prevented the loss of bone induced by RCC RC-9 cells, with a TBV of 12.8 (2.1)%, but the development of hypercalcaemia was unaffected. Olpadronate treatment at a TV of 500 mm(3) did not influence the development of hypercalcaemia and did not protect against bone resorption. Kinetic monitoring showed an identical rate of tumour growth in the presence or absence of bisphosphonate, while under both conditions there was a tumour load-dependent increase in calcium concentration. CONCLUSIONS: Bisphosphonate can prevent parathyroid hormone-related peptide (PTHrP)-mediated bone resorption when administered during the early phase of renal tumour growth, but has no effect on the tumour-induced development of hypercalcaemia, indicating a primary role for renal tubular reabsorption of calcium in the kidney by PTHrP in HHM. PMID- 17428244 TI - Does the molecular staging in pelvic lymph nodes improve the detection of relevant prostate cancer metastases? An assessment after 6 years. AB - OBJECTIVE: To assess the course of cancer-free survival and thus determine how reliably reverse transcriptase-polymerase chain reaction (RT-PCR) can detect prostate-specific antigen (PSA)-expressing cells, as patients with untreated lymph node-positive prostate cancer tend to have a poor prognosis, whereas those treated with radical prostatectomy (RP) and immediate adjuvant hormonal therapy show excellent local disease control and a disease-free survival comparable with that of patients with negative lymph nodes, but the detection of micrometastatic disease in pelvic lymph nodes remains a major challenge. PATIENTS AND METHODS: Quantitative RT-PCR was used to detect PSA mRNA expression in total RNA of 457 pelvic lymph nodes from 70 patients who had RP (53 patients) or laparoscopic lymphadenectomy (17) at our clinic in 1999/2000. For this purpose, alternate sections of lymph node tissue were either snap-frozen for later RNA isolation or examined by standard histopathological methods. Clinicopathological data, adjuvant treatments and follow-up data were recorded for all patients. RESULTS: After January 2006 (6-year observation period), 13 patients had no follow-up data, while 27 had biochemical (PSA) recurrence or other evidence of clinical progression (two died from prostate cancer), and 30 had no signs of recurrence. Compared to the 'reference' standard (histopathology), the PCR method had a sensitivity of 83% and a specificity of 66%. The method had a positive predictive value of 52% and a negative predictive value of 57%. CONCLUSION: Considered alone, pelvic lymph node PSA RT-PCR does not predict the clinical course better than a histopathological assessment of lymph nodes. However, it also identifies some patients with negative histology who later show progression. When added to the pathological classification, PSA RT-PCR improves the detection rate of primary lymphatic dissemination. PMID- 17428245 TI - Roles of attenuated neuronal nitric-oxide synthase protein expression and accelerated arginase activity in impairing neurogenic relaxation of corpus cavernosum in aged rabbits. AB - OBJECTIVE: To investigate whether changes in neuronal nitric oxide synthase (nNOS) protein expression and arginase activity are implicated in impairing the neurogenic cavernosal relaxation in aged rabbits, as NO is important in the neurogenic relaxation of corpus cavernosum during the erectile state. MATERIALS AND METHODS: Cavernosal specimens of young adult (3-6 months old) and aged (36-48 months old) rabbits were used for isometric tension experiments, Western blot analysis, cGMP determination and measurements of NOS and arginase activities. RESULTS: The neurogenic relaxation and cGMP production in response to electrical field stimulation were significantly impaired in aged cavernosal specimens. Western blot analysis showed that nNOS protein was highly expressed in cavernosal specimens from young rabbits, but was undetectable or greatly decreased in old rabbits, with no change in overall NOS activity. Arginase activity in aged cavernosal specimens was significantly higher than in young rabbits. Supplementing with excess l-arginine, or giving S-(2-boronoethyl)-l-cysteine as an arginase inhibitor, significantly increased the neurogenic relaxation at lower frequencies only in the younger rabbits. CONCLUSION: These results suggest that impairment of neurogenic and NO-mediated relaxation in the aged corpus cavernosum possibly results from the down-regulation of nNOS protein. The reduced l-arginine bioavailability to nNOS due to accelerated arginase activity would lead to further impairment of neurogenic NO production, in concert with decreased nNOS protein expression. PMID- 17428246 TI - Long-term functional outcomes of female genital reconstruction in childhood. AB - There are two very interesting paediatric urology papers; in the first, authors from London describe the long-term functional outcomes of female genital reconstruction in childhood, and in the second, authors from Italy present a morphological and manometric study of VUR. PMID- 17428247 TI - Radiotherapy and androgen deprivation in combination after local surgery (RADICALS): a new Medical Research Council/National Cancer Institute of Canada phase III trial of adjuvant treatment after radical prostatectomy. PMID- 17428248 TI - Upper urinary tract tumour after radical cystectomy for transitional cell carcinoma of the bladder: an update on the risk factors, surveillance regimens and treatments. AB - Urothelial carcinoma is characterized by multiple, multifocal recurrences throughout the genitourinary tract; approximately 3% of patients treated by radical cystectomy (RC) for invasive transitional cell carcinoma (TCC) of the bladder will subsequently develop a subsequent TCC in the upper urinary tract (UUT) urothelium. Metachronous upper UUT tumours (mUUT-TCC) typically occur as a late oncological event (>3 years after RC). The vast majority of mUUT-TCCs are detected only after the progression to tumour-related symptoms, e.g. haematuria, flank pain or pyelonephritis, despite strict adherence to surveillance protocols. Failure of imaging and cytology to detect most asymptomatic tumours has led to questions about the need for routine UUT surveillance. Some authors have advocated a more tailored approach to surveillance after RC, targeting high-risk patients and with limiting imaging in those patients at lowest risk of developing a subsequent UUT-TCC. mUUT-TCCs are most common in patients with TCC in the ureter or urethra, and with organ-confined bladder cancer. Although the prognosis is generally poor, long-term survival can be achieved in a subset of patients after radical nephroureterectomy (NU). Minimally invasive techniques, e.g. ureteroscopic and percutaneous resection, have been proposed as renal-sparing alternatives to radical surgery for patients with low-stage and -grade de novo UUT-TCC. However, oncological control of renal-sparing therapies in those with high-risk mUUT-TCC remains largely unconfirmed. Until oncological outcomes equivalent to the standard, radical NU, are reported in patients after RC, conservative treatment strategies should be avoided. PMID- 17428249 TI - 18F-choline and/or 11C-acetate positron emission tomography: detection of residual or progressive subclinical disease at very low prostate-specific antigen values (<1 ng/mL) after radical prostatectomy. AB - OBJECTIVES: To assess the value of positron emission tomography (PET)/computed tomography (CT) with either (18)F-choline and/or (11)C-acetate, of residual or recurrent tumour after radical prostatectomy (RP) in patients with a prostate specific antigen (PSA) level of <1 ng/mL and referred for adjuvant or salvage radiotherapy. PATIENTS AND METHODS: In all, 22 PET/CT studies were performed, 11 with (18)F-choline (group A) and 11 with (11)C-acetate (group B), in 20 consecutive patients (two undergoing PET/CT scans with both tracers). The median (range) PSA level before PET/CT was 0.33 (0.08-0.76) ng/mL. Endorectal-coil magnetic resonance imaging (MRI) was used in 18 patients. Nineteen patients were eligible for evaluation of biochemical response after salvage radiotherapy. RESULTS: There was abnormal local tracer uptake in five and six patients in group A and B, respectively. Except for a single positive obturator lymph node, there was no other site of metastasis. In the two patients evaluated with both tracers there was no pathological uptake. Endorectal MRI was locally positive in 15 of 18 patients; 12 of 19 responded with a marked decrease in PSA level (half or more from baseline) 6 months after salvage radiotherapy. CONCLUSIONS: Although (18)F choline and (11)C-acetate PET/CT studies succeeded in detecting local residual or recurrent disease in about half the patients with PSA levels of <1 ng/mL after RP, these studies cannot yet be recommended as a standard diagnostic tool for early relapse or suspicion of subclinical minimally persistent disease after surgery. Endorectal MRI might be more helpful, especially in patients with a low likelihood of distant metastases. Nevertheless, further research with (18)F choline and/or (11)C-acetate PET with optimal spatial resolution might be needed for patients with a high risk of distant relapse after RP even at low PSA values. PMID- 17428250 TI - Health-insurance status is a determinant of the stage at presentation and of cancer control in European men treated with radical prostatectomy for clinically localized prostate cancer. AB - OBJECTIVE: To determine whether health-insurance status might result in more localized stage at presentation, more favourable stage at surgery and in a lower rate of biochemical recurrence (BCR), in patients diagnosed with prostate cancer and treated with radical prostatectomy (RP), as despite uninhibited access to healthcare, private and public health insurance are available in most European countries. PATIENTS AND METHODS: In all, 4442 consecutive men had RP in two large European centres, of whom 2372 had public and 2070 had private health insurance. The groups were compared for several variables according to insurance status (private vs public). Means and proportions tests were complemented with logistic regression or Kaplan-Meier analyses. RESULTS: Serum prostate-specific antigen level (P < 0.001), clinical stage (P < 0.001), pathological Gleason sum (P = 0.02), positive surgical margin rate (18.4% vs 25.4%, P < 0.001), extracapsular extension rate (17.7% vs 20.0%, P = 0.047) and seminal vesicle invasion rate (9.6% vs 11.6%, P = 0.04) were more favourable in privately insured patients. Conversely, the rate of lymph-node involvement was higher in those with private than public insurance (4.4% vs 3.3%, P = 0.045). In univariate analyses addressing pathological variables, private insurance was invariably protective (all P < 0.05). The Kaplan-Meier analyses showed that privately insured patients had a lower rate of BCR after RP (log-rank P = 0.017). CONCLUSION: Despite uninhibited access to healthcare, insurance status represents a rate-limiting variable, which affects stage at presentation and the outcome of cancer control. PMID- 17428251 TI - Multidetector computed tomography urography for diagnosing upper urinary tract urothelial tumour. AB - OBJECTIVE: To evaluate multidetector computed tomography urography (MDCTU) for diagnosing upper urinary tract (UUT) urothelial tumour by comparison with retrograde ureteropyelography (RUP). PATIENTS AND METHODS: MDCTU and RUP were used in a selected series of adult patients presenting with haematuria. Entry criteria were based on findings on intravenous urography and were chosen to ensure a high prevalence of UUT urothelial tumour to allow a valid retrospective comparison of the diagnostic techniques. MDCTU and RUP studies were scored for the presence and absence of UUT urothelial tumour by two radiologists, retrospectively and independently, and while unaware of the demographic and clinical information. The reference standards were the histopathology and clinical follow-up. RESULTS: MDCTU and RUP were used in 106 patients over a 24 month period. RUP was attempted in 151 of 212 UUTs; the corresponding MDCTU for each UUT was reviewed. MDCTU was a true-positive (TP) for urothelial tumour in 31, true-negative (TN) in 111, false-positive (FP) in eight and false-negative (FN) in one UUT, giving a sensitivity of 0.97, a specificity of 0.93, a positive predictive value (PPV) of 0.79 and a negative PV (NPV) of 0.99. RUP was technically successful and diagnostic in 96% of the UUTs (143/151). For diagnosing urothelial tumour, RUP was TP in 26, TN in 112, FP in four and FN in one UUT, giving a sensitivity of 0.97, specificity of 0.93, a PPV of 0.79 and NPV of 0.99. CONCLUSION: This study validates quantitatively the use of MDCTU for diagnosing UUT urothelial tumour. PMID- 17428252 TI - Diabetes in non-obese diabetic mice is not associated with quantitative changes in CD4+ CD25+ Foxp3+ regulatory T cells. AB - The role of regulatory T cells (Tregs) in maintaining self tolerance has been intensively researched and there is a growing consensus that a decline in Treg function is an important step towards the development of autoimmune diseases, including diabetes. Although we show here that CD25+ cells delay diabetes onset in non-obese diabetic (NOD) mice, we found, in contrast to previous reports, neither an age-related decline nor a decline following onset of diabetes in the frequency of CD4+ CD25+ Foxp3+ regulatory T cells. Furthermore, we demonstrate that CD4+ CD25+ cells from both the spleen and pancreatic draining lymph nodes of diabetic and non-diabetic NOD mice are able to suppress the proliferation of CD4+ CD25- cells to a similar extent in vitro. We also found that pretreatment of NOD mice with anti-CD25 antibody allowed T cells with a known reactivity to islet antigen to proliferate more in the pancreatic draining lymph nodes of NOD mice, regardless of age. In addition, we demonstrated that onset of diabetes in NOD.scid mice is faster when recipients are co-administered splenocytes from diabetic NOD donors and anti-CD25. Finally, we found that although diabetic CD4+ CD25+ T cells are not as suppressive in cotransfers with effectors into NOD.scid recipients, this may not indicate a decline in Treg function in diabetic mice because over 10% of CD4+ CD25+ T cells are non-Foxp3 and the phenotype of the CD25- contaminating population significantly differs in non-diabetic and diabetic mice. This work questions whether onset of diabetes in NOD mice is associated with a decline in Treg function. PMID- 17428253 TI - Telomere dysfunction and inactivation of the p16(INK4a)/Rb pathway in pyothorax associated lymphoma. AB - Previous studies have indicated that genome instability is involved in the lymphomagenesis of pyothorax-associated lymphoma (PAL), which develops in patients with a long-standing history of pyothorax. One of the well-known causes of genome instability is telomere dysfunction. In the present study, the condition of telomeres was analyzed in the cell lines and clinical samples from PAL. Telomere length (TL) in PAL cell lines was extremely short (<4.5 kbp). TL in tumor samples was broad in range, and shorter than that in the peripheral blood leukocytes from the matched patients. Three of five PAL cell lines showed frequent loss of telomere signals (telomere erosion); however, telomerase activity in PAL cell lines was similar to that in Burkitt lymphoma cell lines. Rb expression was detected in three PAL cell lines and four of 15 clinical samples, respectively. Rb protein expressed in three PAL cell lines was heavily phosphorylated, indicating that function of Rb protein was suppressed. p16(INK4a) expression was not detected in either cell lines or clinical samples. The promoter region in p16(INK4a) was heavily methylated in all cell lines as well as the clinical samples. Inactivation of the p16(INK4a)/Rb pathway may allow continuous cell division and critical telomere shortening, which induce genome instability, finally leading to malignant transformation. Taken together, telomere dysfunction and inactivation of the p16(INK4a)/Rb pathway might play a role for PAL development. PMID- 17428254 TI - Combined treatment with green tea catechins and sodium nitrite selectively promotes rat forestomach carcinogenesis after initiation with N-methyl-N'- nitro N-nitrosoguanidine. AB - Combined treatment with several phenolic antioxidants and sodium nitrite (NaNO(2)) has already shown to enhance rat forestomach carcinogenesis. In the present experiment, effects of green tea catechins (GTC) alone or in combination with NaNO(2) on gastric carcinogenesis were investigated in a rat two-stage carcinogenesis model. Groups of eight, 6-week-old F344 male rats were given 0.01%N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in their drinking water and 5% NaCl in the diet for 10 weeks for glandular stomach initiation and a single intragastric administration of 100 mg/kg/bodyweight of MNNG at week 9 for forestomach initiation. From week 11, they received either drinking water containing 0.2% NaNO(2) and a diet supplemented with 1% GTC in combination, each individual chemical alone or a basal diet until the end of week 42. In the forestomach, incidences and multiplicities of neoplastic lesions were clearly increased by the combined treatment, in spite of GTC alone suppressing the occurrence of papillomas. In a short-term experiment with similar protocol without MNNG pretreatment, a significant increase of 8-hydroxydeoxyguanosine (8 OHdG) levels in forestomach DNA occurred 24 h after the combined treatment, concomitant with erosion and inflammatory cell infiltration. In an in vitro study, electron spin resonance demonstrated hydroxyl radical formation after incubation of epigallocatechin gallate or epicatechin gallate with the NO generator, NOC-7. Thus, GTC alone showed a weak chemopreventive effect on forestomach carcinogenesis, but in the presence of NaNO(2) it exerted a promotive effect which might involve hydroxyl-radical-associated oxidative DNA damage. However, no influence was exerted in the glandular stomach. PMID- 17428255 TI - Expression and DNA methylation patterns of Tslc1 and Dal-1 genes in hepatocellular carcinomas induced by N-nitrosodiethylamine in rats. AB - To assess the involvement of the TSLC cascade in hepatocarcinogenesis, we investigated the expression and DNA methylation patterns of the genes Tslc1 and Dal-1 in hepatocellular carcinomas (HCC) induced using N-nitrosodiethylamine (DEN) in rats. Six-week-old male F344 rats received a single intraperitoneal injection of DEN at a dose of 10 mg/kg body weight, followed by combined treatment with partial hepatectomy and colchicine to induce cell-cycle disturbance and a selection procedure consisting of 2-acetylaminofluorene and carbon tetrachloride. Total RNA was extracted from 10 HCC, and the expression levels of Tslc1 and Dal-1 were measured using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis. Three of 10 HCC showed reduced expression of Tslc1, compared with normal liver tissues, but no changes in the expression level of Dal-1 were found. For DNA methylation analysis, bisulfite sequencing was performed. The 5' upstream region of Tslc1 was methylated in the three HCC in which its expression was reduced, but was unmethylated in normal liver tissue. Western blot analysis also revealed reduced expression of Tslc1 protein in the three HCC. These results suggest that alterations to the TSLC cascade might have a role in hepatocarcinogenesis using DEN in rats. PMID- 17428256 TI - Fully human antibody exhibits pan-human leukocyte antigen-DR recognition and high in vitro/vivo efficacy against human leukocyte antigen-DR-positive lymphomas. AB - HD8, a fully human monoclonal antibody specific for human leukocyte antigen-DR (HLA-DR), was generated by using the transchromosome mouse that bears the human immunoglobulin genes. HD8 could bind to all 13 tested HLA-DR-positive cell lines and 35 B-cells from healthy donors. Epitope mapping revealed that while the antibody recognizes the most polymorphic region of the HLA-DRB chain, its critical epitope residues are conserved in the major alleles. Indeed, HD8 could recognize 99.2% of HLA-DRB alleles. Since its essential epitope residues are also largely conserved in HLA-DP and HLA-DQ, HD8 could recognize 100% and 66% of the HLA-DP and HLA-DQ alleles tested, respectively. HD8 exerted strong antibody dependent cellular cytotoxicity and complement-dependent cytotoxicity in vitro, and significantly extended the life span of immunocompromised mice inoculated with non-Hodgkin lymphoma cell lines. The HD8 antibody may be highly useful in HLA-DR-targeted immunotherapy as it is likely to evoke similarly strong responses in individuals carrying different HLA-DR alleles. PMID- 17428257 TI - von Willebrand factor type D domain mutant of SVS-1/SUSD2, vWD(m), induces apoptosis in HeLa cells. AB - SVS-1/SUSD2 is a novel gene, which inhibits growth and reverses tumorigenic phenotypes of cancer cells in vitro. Here we report identification of a mutant of SVS-1, designated SVS-1-vWD(m), in which conserved amino acids GLLG at positions 591-594 in von Willebrand factor type D (vWD) domain are replaced by AAAA. As observed by laser confocal microscope, intracellular localization of the mutant protein has changed such that both the N-terminus and the C-terminus of SVS-1 vWD(m) were localized in the inner surface of the plasma membrane, whereas the N terminus of SVS-1 was localized in the outer surface of the plasma membrane. Additionally, SVS-1-vWD(m) was processed much less efficiently and in a slightly different manner. In in vitro studies, adenovirus-mediated transduction of the SVS-1-vWD(m)gene induced growth suppression of HeLa cells in a dose-dependent manner, as the wild-type gene and inhibition of anchorage-independent growth. Of great interest is the finding that the mutant protein, vWD(m), but not the wild type one induced apoptosis, as observed by nuclear as well as DNA fragmentation. Activation of caspase-3 and -9, but not caspase-8 or -12, was also demonstrated in vWD(m)-expressing cells. An inhibition of Akt phosphorylation, a major survival signaling component, also occurred in vWD(m)-expressing HeLa cells. Together these data suggest that vWD(m) induces apoptosis by inactivation of survival signaling component Akt and activation of caspase cascade (mitochondrial pathway) in HeLa cells. We propose SVS-1-vWD(m)as an alternative gene for use in developing new therapeutic strategies for the treatment of cancer. PMID- 17428258 TI - Isolation of a novel mouse gene, mSVS-1/SUSD2, reversing tumorigenic phenotypes of cancer cells in vitro. AB - We report isolation of a novel tumor-reversing gene, tentatively named SVS-1, encoding a protein of 820 amino acids with localization on the plasma membrane as a type I transmembrane protein. The gene was found among those downregulated in the activated oncogene-v-K-ras-transformed NIH3T3 cells, Ki3T3, with tumorigenic phenotype. SVS-1 protein harbors several functional domains inherent to adhesion molecules. Histochemical staining of mouse tissues using antibody raised against the protein showed the expression of the protein in restricted regions and cells, for example, strongly positive in apical membranes of epithelial cells in renal tubules and bronchial tubes. The protein inducibly expressed in human fibrosarcoma HT1080 cells and cervical carcinoma HeLa cells was found to be localized primarily on the plasma membrane, as stained with antibodies against FLAG tag in the N-terminus and against the C-terminal peptide of the protein. Expression of the protein in cells induced a variety of biological effects on cancer cells: detachment from the substratum and aggregation of cells and growth inhibition in HeLa cells, but no inhibition in non-tumorigenic mouse NIH3T3 cells. Inhibition of clonogenicity, anchorage-independent growth, migration and invasion through Matrigel was also observed. Taken together these results suggest that the SVS-1 gene is a possible tumor-reversing gene. PMID- 17428259 TI - Association of maternal fat and alcohol intake with maternal and umbilical hormone levels and birth weight. AB - High levels of estrogen during pregnancy have been hypothesized to increase the risk of breast cancer in offspring. Some studies have reported a positive association of estrogen level during pregnancy with fetal size, which has been linked to the subsequent risk of breast cancer in offspring. We examined whether maternal diet, including fat and alcohol intake, was associated with hormone levels during pregnancy, as well as with birth weight. The concentrations of estradiol, estriol, and testosterone were measured in the maternal serum and umbilical cord blood of 189 women during pregnancy and at delivery. Intakes of fat, alcohol, and other nutrients were assessed by 5-day diet records at approximately the 29th week of pregnancy before blood sampling. Intake of polyunsaturated fatty acids was moderately but significantly positively correlated with the umbilical cord estriol level (r = 0.17, P = 0.03) after controlling for covariates. The positive association between intake of polyunsaturated fatty acids and birth weight was of borderline significance (r = 0.14, P = 0.06). Intake of long-chain n-3 fatty acids was significantly inversely correlated with the umbilical cord estradiol and testosterone levels (r = -0.18, P = 0.02 and r = -0.24, P = 0.002, respectively). Alcohol intake was significantly positively correlated with the maternal estradiol level in the 29th week of pregnancy (r = 0.19, P = 0.01), but was unrelated to birth weight. Estrogen level during pregnancy may be regulated by dietary polyunsaturated fatty acids and mediate their effects on fetal growth. PMID- 17428260 TI - Improving the efficacy of trastuzumab in breast cancer. AB - Although overexpression of human epidermal growth factor receptor 2 (HER2) protein, amplification of the gene or both are associated with poor prognosis in breast cancer, trastuzumab has clearly provided clinical benefits in metastatic breast cancer, adjuvant treatment settings and primary systemic therapy. However, even in those HER2 overexpressors, the majority of patients who achieve an initial response generally acquire resistance within 1 year. Therefore, it is critical to elucidate the mechanism of resistance and to search for better combination treatments with chemotherapeutic agents or other novel modalities. Here, we discuss both clinical and preclinical data regarding these issues. PMID- 17428261 TI - Simvastatin inactivates beta1-integrin and extracellular signal-related kinase signaling and inhibits cell proliferation in head and neck squamous cell carcinoma cells. AB - The 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase inhibitors, also called statins, are commonly used as lipid-lowering drugs that inhibit cholesterol biosynthesis. An anticancer effect, as a pleiotropic function of certain statins, has been hypothesized. In the present study, we investigated the effect of simvastatin, one of the natural statins, on cell proliferation, cell cycle, invasive activity, and molecular expressions associated with cell-extracellular matrix adhesion, signal transduction, and DNA synthesis in Tu167 and JMAR cells from head and neck squamous cell carcinoma. The addition of simvastatin resulted in a dose-dependent inhibition of cell growth and migration into the extracellular matrix. Considerable morphological changes occurred after treatment with simvastatin, demonstrating loss of cell adhesion and disruption of actin filaments in cytoplasm. The inhibitory effect of simvastatin on cell proliferation seemed to be associated with cell cycle arrest and increased expression of p21, p27, and activated caspase-3. The expression of beta1 integrin, a counter adhesion for the extracellular matrix, phosphorylated FAK, and phosphorylated ERK was decreased by treatment with simvastatin. The proapoptotic effect of simvastatin was inhibited by treatment with mevalonate. cDNA microarray assay demonstrated that molecular changes resulting from treatment with simvastatin included the up-regulation of cell cycle regulators and apoptosis-inducing factors and the down-regulation of integrin-associated molecules and cell proliferation markers. Of down-regulated genes induced by simvastatin treatment, a significant depletion of thymidylate synthase was confirmed using western blot analysis. These results imply that simvastatin has the potential to be effective for the prevention of the growth and metastasis of cancer cells. PMID- 17428262 TI - Characterization and expression of a maternal axolotl cyclin B1 during oogenesis and early development. AB - The M phase promoting factor (MPF) is a dimer composed of a catalytic Cdk1 subunit and a Cyclin B regulatory subunit. We have characterized a cDNA containing the entire coding sequence of an axolotl Cyclin B1 protein that is able to promote MPF activity when added to a fraction from prophase I oocytes that contains monomeric Cdk1. The axolotl cyclin B1 gene is expressed as a maternal mRNA in oocytes and early embryos. Its poly(A) tail length increases in metaphase II oocytes and then decreases regularly during the first embryonic cell cycles. Endogenous Cyclin B1 protein is first expressed during oocyte meiotic maturation. Its level oscillates after fertilization and is coordinated to the phosphorylation level of tyrosine 15 residue of Cdk1 (pTyr15), with both maxima preceding each cell division. As expected, when translated into microinjected oocytes, axolotl Cyclin B1 induces the resumption of meiosis. In electrically activated unfertilized eggs (UFE), Cyclin B1 and pTyr15 cyclic accumulations are observed with kinetics different from those of the early embryonic cycles. The axolotl embryo and UFE provide interesting in vivo comparative models for studying events controlling Cyclin B1 regulation during development. PMID- 17428263 TI - Regional specification of the endoderm in the early chick embryo. AB - In the avian embryo, the endoderm, which forms a simple flat-sheet structure after gastrulation, is regionally specified in a gradual manner along the antero posterior and dorso-ventral axes, and eventually differentiates into specific organs with defined morphologies and gene expression profiles. In our study, we carried out transplantation experiments using early chick embryos to elucidate the timing of fate establishment in the endoderm. We showed that at stage 5, posteriorly grafted presumptive foregut endoderm expressed CdxA, a posterior endoderm marker, but not cSox2, an anterior endoderm marker. Conversely, anteriorly grafted presumptive mid-hindgut endoderm expressed cSox2 but not CdxA. At stage 8, posteriorly grafted presumptive foregut endoderm also expressed CdxA and not cSox2, but anteriorly grafted presumptive mid-hindgut endoderm showed no changes in its posterior-specific gene expression pattern. At stage 10, both posteriorly grafted foregut endoderm and anteriorly grafted mid-hindgut endoderm maintain their original gene expression patterns. These results suggest that the regional specification of the endoderm occurs between stages 8 and 10 in the foregut, and between stages 5 and 8 in the mid-hindgut. PMID- 17428264 TI - L-Maf regulates p27kip1 expression during chick lens fiber differentiation. AB - Organ formation requires spatio-temporal proliferation and differentiation of precursor cells. During lens development, placodal cells in the posterior lens vesicle exit from the cell cycle and enter into the process of differentiation. Cyclin-dependent kinase inhibitors play critical roles in cell cycle exit and promote differentiation in several tissues. We have found that p27kip1 is expressed in the posterior lens cells that undergo differentiation to form the differentiated fiber cells. The transcription factor L-Maf is expressed in these cells earlier than p27kip1. From in ovo gain- or loss-of-function experiments, we have found that L-Maf can, respectively, induce or inhibit the expression of p27kip1 in lens cells. Promoter assays using the 5' upstream sequences of the human p27kip1 gene indicate that L-Maf can activate p27kip1 transcription through the basal regulatory region. We suggest that L-Maf regulates cell cycle exit of the posterior lens cells by activating p27kip1, and thus directs fiber cell differentiation during lens formation in chick. PMID- 17428265 TI - Functional expression of connexin30 and connexin31 in the polarized human airway epithelium. AB - Gap junctions are documented in the human airway epithelium but the functional expression and molecular identity of their protein constituents (connexins, Cx) in the polarized epithelium is not known. To address this question, we documented the expression of a family of epithelial Cx (Cx26, Cx30, Cx30.3, Cx31, Cx31.1, Cx32, Cx37, Cx40, and Cx43) in primary human airway epithelial cells (AEC) grown on porous supports. Under submerged conditions, AEC formed a monolayer of airway cells whereas the air-liquid interface induced within 30-60 days AEC differentiation into a polarized epithelium for up to 6-9 months. Maturation of AEC was associated with the down-regulation of Cx26 and Cx43. The well differentiated airway epithelium exhibited gap junctional communication between ciliated and between ciliated and basal cells. Interestingly, Cx30 was mostly present between ciliated cells whereas Cx31 was found between basal cells. These results are supportive of the establishment of signal-selective gap junctions with maturation of AEC, likely contributing to support airway epithelium function. These results lay the ground for studying the role of Cx-mediated cell cell communication during repair following AEC injury and exploring Cx-targeted interventions to modulate the healing process. PMID- 17428266 TI - Clock genes beyond the clock: CLOCK genotype biases neural correlates of moral valence decision in depressed patients. AB - Gene polymorphisms in the mammalian biological clock system influence individual rhythms. A single nucleotide polymorphism (SNP) in the 3' flanking region of CLOCK (3111 T/C; rs1801260) influenced diurnal preference in healthy humans and caused sleep phase delay and insomnia in patients affected by bipolar disorder. Genes of the biological clock are expressed in many brain structures other than in the 'master clock' suprachiasmatic nuclei. These areas, such as cingulate cortex, are involved in the control of many human behaviors. Clock genes could then bias 'nonclock' functions such as information processing and decision making. Thirty inpatients affected by a major depressive episode underwent blood oxygen-level dependent (BOLD) functional magnetic resonance imaging (fMRI). The cognitive activation paradigm was based on a go/no-go task. Morally connoted words were presented. Genotyping of CLOCK was performed for each patients. We measured activity levels through actimetry during the day before the fMRI study. CLOCK 3111 T/C SNP was associated with activity levels in the second part of the day, neuropsychological performance and BOLD fMRI correlates (interaction of genotype and moral valence of the stimuli). Our results support the hypothesis that individual clock genotype may influence several variables linked with human behaviors in normal and psychopathological conditions. PMID- 17428267 TI - Downregulation of mu opioid receptor by RNA interference in the ventral tegmental area reduces ethanol consumption in mice. AB - Pharmacological and genetic studies have implicated the mu opioid receptor (MOR) in the regulation of ethanol intake in animal models and humans. Non-specific antagonists of opioid receptors have been shown to affect ethanol consumption when infused directly into the ventral tegmental area (VTA) of rats. However, administration of MOR-selective antagonists into the VTA has yielded mixed results. We used RNA interference (RNAi) to specifically decrease levels of MOR messenger RNA in the VTA of mice and examined the effect on ethanol consumption in a two-bottle choice paradigm. Mice were injected in the VTA with lentivirus expressing either a small hairpin RNA (shRNA) targeting MOR or a control shRNA. One week after virus injection, mice were examined for ethanol consumption in a two-bottle choice experiment with increasing concentrations of ethanol over the course of 1 month. Expression of an shRNA targeting MOR in the VTA led to a significant reduction in ethanol consumption. These results strengthen the hypothesis that MOR in the VTA is one of the key brain substrates mediating alcohol consumption. The RNAi combined with lentiviral delivery can be used successfully in brain to effect a sustained reduction in expression of specific genes for behavioral analysis. PMID- 17428268 TI - The effect of the baby-friendly hospital initiative on long-term breast feeding. AB - The aim of this study was to evaluate the effects of 'baby-friendly hospital initiative' (BFHI) on breast feeding. In the four consecutive months after BFHI in Gazi University Hospital (November 2002-February 2003), breast feeding status until the second year of life in 297 babies, born in the same hospital was compared with the values of 258 babies born before BFHI (November 2001-February 2002). The exclusive breast feeding rate in the first 6 months was higher in the babies born after BFHI. Cox regression analysis revealed that BFHI increases the duration of breast feeding 1.5 times. At the end of the second year, cumulative rate of breast feeding was higher in the group after-BFHI (p=0.0036). The rate of breast feeding was increased by BFHI implementation. PMID- 17428269 TI - Annihilation of the sinus node. PMID- 17428270 TI - Time course of esophageal lesions after catheter ablation with cryothermal and radiofrequency ablation: implication for atrio-esophageal fistula formation after catheter ablation for atrial fibrillation. AB - BACKGROUND: Atrio-esophageal fistulas have been described as a consequence of radiofrequency (RF) ablation for the treatment of atrial fibrillation (AF). However, whether cryoablation can avoid this potential fatal complication remains unclear. METHODS AND RESULTS: We studied the effects of direct application of RF and cryoablation on the cervical esophagus in 16 calves. Cryoablation was performed with a 6.5-mm catheter probe using a single 5-minute freeze at <-80 degrees C, and RF ablation was delivered with an 8-mm catheter electrode at 50 W and 50 degrees C for 45-60 seconds. Histopathologic assessments were performed at 1, 4, 7, and 14 day(s) after completion of the ablation protocol: four animals were examined each day. A total of 85 direct esophageal ablations were performed: 41 with RF and 44 with cryoablation. There were no significant differences in lesion width, depth, or volume between cryoablation and RF ablation at Day 1, 4, and 14 after the procedure (P > 0.05). However, lesion width and volume were significantly larger with RF than with cryoablation at Day 7. Although acute (Day 1) and chronic (Day 14) RF and cryoablation lesions were of comparable size, histologic evidence of partial- to full-wall esophageal lesion ulceration was observed in 0 of 44 (0%) lesions with cryoablation, compared with 9 of 41 (22%) lesions with RF ablation (P = 0.0025). CONCLUSIONS: Direct application of cryoablation and RF ablation created similar acute and chronic lesion dimensions on the esophagus. However, cryoablation was associated with a significantly lower risk of esophageal ulceration, compared with RF ablation. PMID- 17428271 TI - Incidence of atrial arrhythmias detected by permanent pacemakers (PPM) post pulmonary vein antrum isolation (PVAI) for atrial fibrillation (AF): correlation with symptomatic recurrence. AB - BACKGROUND: Studies examining AF recurrences post-PVAI base recurrence on patient reporting of symptoms. However, whether asymptomatic recurrences are common is not well known. OBJECTIVE: To assess the incidence of atrial tachycardia/fibrillation post-PVAI as detected by a PPM and whether these recurrences correlate to symptomatic recurrence. METHODS: Eighty-six consecutive patients with symptomatic AF and PPMs with programmable mode-switch capability underwent PVAI. Mode switching was programmed post-PVAI to occur at an atrial sensed rate of >170 bpm. Patients were followed with clinic visits, ECG, and PPM interrogation at 1, 3, 6, and 9 months post-PVAI. The number and duration of mode switching episodes (MSEs) were recorded at each visit and is presented as median (interquartile range). RESULTS: The patients (age 57 +/- 8 years, EF 54 +/- 10%) had paroxysmal (65%) and persistent (35%) AF pre-PVAI. Sensing, pacing, and lead function were normal for all PPMs at follow-up. Of the 86 patients, 20 (23%) had AF recurrence based on symptoms. All 20 of these patients had appropriate MSEs detected. Of the 66 patients without symptomatic recurrence, 21 (32%) had MSEs detected. In 19 of these patients, MSEs were few in number, compared with patients with symptomatic recurrence (16 [4-256] vs 401 [151-2,470], P < 0.01). The durations were all <60 seconds. All of these nonsustained MSEs occurred within the first 3 months post-PVAI, gradually decreasing over time. The other 2 of 21 remaining patients had numerous (1,343 [857-1,390]) and sustained (18 +/- 12 minutes) MSEs that also persisted beyond 3 months (1 beyond 6 months). Therefore, the incidence of numerous, sustained MSEs in asymptomatic patients post-PVAI was 2 of 66 (3%). CONCLUSIONS: Detection of atrial tachyarrhythmias by a PPM occurred in 30% of patients without symptomatic AF recurrence. Most of these episodes were <60 seconds and waned within 3 months. Sustained, asymptomatic episodes were uncommon. PMID- 17428272 TI - Avoidance of right ventricular pacing in cardiac resynchronization therapy improves right ventricular hemodynamics in heart failure patients. AB - BACKGROUND: Cardiac resynchronization therapy (CRT) applied by pacing the left and right ventricles (BiV) has been shown to provide synchronous left ventricular (LV) contraction in heart failure patients. CRT may also be accomplished through synchronization of a properly timed LV pacing impulse with intrinsically conducted activation wave fronts. Elimination of right ventricular (RV) pacing may provide a more physiological RV contraction pattern and reduce device current drain. We evaluated the effects of LV and BiV pacing over a range of atrioventricular intervals on the performance of both ventricles. METHODS: Acute LV and RV hemodynamic data from 17 patients with heart failure (EF = 30 +/- 1%) and a wide QRS (138 +/- 25 msec) or mechanical dyssynchrony were acquired during intrinsic rhythm, BiV, and LV pacing. RESULTS: The highest LV dP/dt(max) was achieved during LV pre- (LV paced prior to an RV sense) and BiV pacing, followed by that obtained during LV post-pacing (LV paced after an RV sense) and the lowest LV dP/dt(max) was recorded during intrinsic rhythm. Compared with BiV pacing, LV pre-pacing significantly improved RV dP/dt(max) (378 +/- 136 mmHg/second vs 397 +/- 136 mmHg/second, P < 0.05) and preserved RV cycle efficiency (61.6 +/- 14.6% vs 68.6 +/- 11.4%, P < 0.05) and stroke volume (6.6 +/ 4.4 mL vs 9.0 +/- 6.3 mL, P < 0.05). Based on LV dP/dt(max), the optimal atrioventricular interval could be estimated by subtracting 30 msec from the intrinsic atrial to sensed RV interval. CONCLUSIONS: Synchronized LV pacing produces acute LV and systemic hemodynamic benefits similar to BiV pacing. LV pacing at an appropriate atrioventricular interval prior to the RV sensed impulse provides superior RV hemodynamics compared with BiV pacing. PMID- 17428273 TI - Implantable cardioverter-defibrillator shocks increase T-wave alternans. AB - INTRODUCTION: While implantable defibrillator shocks save lives, shock can lead to ventricular arrhythmias. However, the mechanism of shock-related proarrhythmia remains unclear. We evaluated the impact of ICD shock on repolarization instability, a factor associated with ventricular arrhythmogenesis. METHODS AND RESULTS: Sixty-five patients with ICDs underwent ambulatory ECG monitoring during defibrillation testing 3 months postimplant. TWA was analyzed continuously in the time domain during baseline, sedated, and post-shock states. RR, QRS, and QT intervals and catecholamines were also measured continuously. Adequate pre- and post-shock Holter data were recorded in 55 patients, 48 male, mean 64 +/- 12 years, 50 with coronary disease, 48 with prior spontaneous or induced arrhythmia. TWA significantly increased after shock, from 9.6 +/- 0.5 to 11.9 +/- 0.6 microV, as did QRS duration, epinephrine, and norepinephrine levels, compared with sedated and baseline states. RR intervals decreased minimally. TWA changes with shock were not associated with RR or QRS duration changes, but were associated with changes in epinephrine. CONCLUSIONS: ICD shock, even in the sedated state, increases repolarization instability as measured by TWA, an effect mediated in part by sympathetic stimulation. This association between shock and TWA may have important mechanistic and clinical implications for optimization of defibrillation therapy. PMID- 17428274 TI - Successful control of Scedosporium prolificans septic arthritis and probable osteomyelitis without radical surgery in a long-term renal transplant recipient. AB - Scedosporium species are increasingly isolated from immunocompromised and immunocompetent patients. Scedosporium infections are generally resistant to multiple antifungals, and Scedosporium prolificans is particularly resistant to all single antifungal agents currently in use with in vitro testing. We report here a long-term renal transplant recipient who developed isolated S. prolificans septic monoarthritis and probable osteomyelitis. The infection was successfully treated with a combination of voriconazole and terbinafine in addition to joint washout but did not require radical surgery. This combination has been shown to have synergistic in vitro effect, and anecdotal in vivo success has also been reported recently. We also review the clinical presentation, treatment, and outcome of S. prolificans infection in patients with solid organ transplantation. PMID- 17428275 TI - Development of Kaposi's sarcoma under sirolimus-based immunosuppression and successful treatment with imiquimod. AB - Kaposi's sarcoma (KS) is a vascular neoplasm typically observed in the immunocompromised patient populations, such as acquired immunodeficiency syndrome or transplant patients. KS can appear simultaneously at multiple sites as red to purple, maculo-papular or nodular cutaneous lesions sometimes showing a visceral extension. Sirolimus, an immunosuppressive agent with potent antitumor activity, has been effective in combating post-transplant KS. However, an aggressive regimen of immunosuppression for therapy of severe acute rejection episodes may abolish the antitumor effects of sirolimus. The following is a description of KS development under immunosuppressive therapy with sirolimus, and the successful treatment of KS lesions utilizing the topical application of imiquimod 5% cream, an immune response modifier. PMID- 17428276 TI - Suppression of cell-mediated immunity by a donor-transmitted lymphocytic choriomeningitis virus in a kidney transplant recipient. AB - Infection with lymphocytic choriomeningitis virus (LCMV) in rodents, the primary host, is known to cause suppression of cell-mediated immunity. Serial determinations using a functional cell-mediated immune assay in a kidney transplant recipient with donor-transmitted LCMV also suggested profound suppression of cellular immunity. This suppression persisted in spite of reduction of immunosuppression. With the clearance of the virus there was reconstitution of the cellular immune response. PMID- 17428277 TI - Rhodococcus lung abscess complicating kidney transplantation: successful management by combination antibiotic therapy. AB - In this report, a renal transplant recipient with Rhodococcus lung abscess is described. A high clinical suspicion and appropriate combination antibiotic therapy obviated the need for surgical intervention and was associated with a good clinical outcome. The optimal regimen of combination antibiotic therapy is discussed. PMID- 17428278 TI - Blastomycosis in solid organ transplant recipients. AB - BACKGROUND: Blastomyces dermatitidis, the etiologic agent of blastomycosis, causes severe disease and substantial mortality in those immunocompromised by acquired immunodeficiency syndrome or malignancy. In solid organ transplant recipients, the epidemiology, clinical features, and outcomes have not been fully described. METHODS: We conducted a retrospective case-series at the University of Wisconsin Hospital and Clinics. Case patients were solid organ transplant recipients with blastomycosis. RESULTS: From 1986 to 2004, we identified 11 cases of post-transplant blastomycosis with 64% occurring between 2000 and 2004. Onset of infection occurred a median of 26 months post transplantation with near equal distribution before and after the first year of transplantation. Rejection did not precede any case of post-transplant blastomycosis. Opportunistic co infections were common, occurring in 36% of patients. Pneumonia was the most common clinical presentation and was frequently complicated by acute respiratory distress syndrome (ARDS). Extrapulmonary disease predominantly involved the skin and spared the central nervous system. The overall mortality rate was 36%; however, this increased to 67% in those with ARDS. None of the surviving patients relapsed or received routine secondary antifungal prophylaxis. CONCLUSION: Blastomycosis is an uncommon infection following solid organ transplantation that is frequently complicated by ARDS, dissemination, and opportunistic co-infection. After cure, post-transplant blastomycosis may not require lifelong antifungal suppression. PMID- 17428279 TI - Cerebral toxoplasmosis in an allogeneic peripheral stem cell transplant recipient: case report and review of literature. AB - We present a patient who underwent allogeneic peripheral stem cell transplantation (PSCT) for chronic myelocytic leukemia. Twenty months after the PSCT he experienced status epilepticus. Magnetic resonance imaging (MRI) revealed a focus in the ventral thalamus-hypothalamus region. Using stereotactic biopsy with histology and specific polymerase chain reaction investigation from brain tissue, cerebral toxoplasmosis was diagnosed and treated with antiparasitic therapy. Early recognition of such serious and potentially lethal disease enabled prompt specific treatment. This case report emphasizes the role of stereotactic biopsy in diagnosis of cerebral toxoplasmosis. Other methods such as MRI are non invasive but not sufficiently specific and sensitive. PMID- 17428280 TI - Case of Staphylococcus schleiferi subspecies coagulans endocarditis and metastatic infection in an immune compromised host. AB - Staphylococcus schleiferi subspecies coagulans is a relatively recently described organism with limited human pathogenicity. Described case reports of infection are primarily limited to canine external otitis with only one reported case of human involvement, localized to a surgical wound infection. Similarities in phenotypic and microbiological characteristics between S. schleiferi subsp. coagulans and S. aureus have resulted in frequent mistakes using common methods of identification. We describe the first case of disseminated human infection with this species, definitively identified using DNA sequencing, and successfully treated with conventional therapy. PMID- 17428282 TI - Trends in organ donation and transplantation in the United States, 1996-2005. PMID- 17428283 TI - Organ donation and utilization in the United States, 1996-2005. AB - The success of clinical transplantation as a therapy for end-stage organ failure is limited by the availability of suitable organs for transplant. This article discusses continued efforts by the transplant community to collaboratively improve the organ supply. There were 7593 deceased organ donors in 2005. This represents an all-time high and a 6% increase over 2004. Increases were noted in deceased organ donation of all types of organs; notable is the increase in lung donation, which occurred in 17% of all deceased donors. The percentage of deceased donations that occurred following cardiac death has also reached a new high at 7%. The number of living donors decreased by 2%, from 7003 in 2004 to 6895 in 2005. This article discusses the continued efforts of the Organ Donation Breakthrough Collaborative and the Organ Transplantation Breakthrough Collaborative to support organ recovery and use and to encourage the expectation that for every deceased donor, all organs will be placed and transplanted. PMID- 17428284 TI - Pediatric transplantation in the United States, 1996-2005. AB - Solid organ transplantation is accepted as a standard lifesaving therapy for end stage organ failure in children. This article reviews trends in pediatric transplantation from 1996 to 2005 using OPTN data analyzed by the Scientific Registry of Transplant Recipients. Over this period, children have contributed significantly to the donor pool, and although the number of pediatric donors has fallen from 1062 to 900, this still accounts for 12% of all deceased donors. In 2005, 2% of 89,884 candidates listed for transplantation were less than 18 years old; in 2005, 1955 children, or 7% of 28,105 recipients, received a transplant. Improvement in waiting list mortality is documented for most organs, but pretransplant mortality, especially among the youngest children, remains a concern. Posttransplant survival for both patients and allografts similarly has shown improvement throughout the period; in most cases, survival is as good as or better than that seen in adults. Examination of immunosuppressive practices shows an increasing tendency across organs toward tacrolimus-based regimens. In addition, use of induction immunotherapy in the form of anti-lymphocyte antibody preparations, especially the interleukin-2 receptor antagonists, has increased steadily. Despite documented advances in care and outcomes for children undergoing transplantation, several considerations remain that require attention as we attempt to optimize transplant management. PMID- 17428285 TI - Kidney and pancreas transplantation in the United States, 1996-2005. AB - Kidney and pancreas transplantation in 2005 improved in quantity and outcome quality, despite the increasing average age of kidney graft recipients, with 56% aged 50 or older. Geography and ABO blood type contribute to the discrepancy in waiting time among the deceased donor (DD) candidates. Allocation policy changes are decreasing the median times to transplant for pediatric recipients. Overall, 6% more DD kidney transplants were performed in 2005 with slight increases in standard criteria donors (SCD) and expanded criteria donors (ECD). The largest increase (39%) was in donation after cardiac death (DCD) from non-ECD donors. These DCD, non-ECD kidneys had equivalent outcomes to SCD kidneys. 1-, 3- and 5 year unadjusted graft survival was 91%, 80% and 70% for non-ECD-DD transplants, 82%, 68% and 53% for ECD-DD grafts, and 95%, 88% and 80% for living donor kidney transplants. In 2005, 27% of patients were discharged without steroids compared to 3% in 1999. Acute rejection decreased to 11% in 2004. There was a slight increase in the number of simultaneous pancreas-kidney transplants (895), with fewer pancreas after kidney transplants (343 from 419 in 2004), and a stable number of pancreas alone transplants (129). Pancreas underutilization appears to be an ongoing issue. PMID- 17428286 TI - Liver and intestine transplantation in the United States, 1996-2005. AB - The number of liver transplants performed yearly has slowly and steadily increased over the last 10 years, reaching 6441 procedures in 2005. The number of living donor liver transplants performed rose steadily from 1996 to 2001, when it peaked at 519; since 2003 there have been approximately 320 such procedures performed each year. The continual increase in the size of the waiting list for a liver transplant, which peaked in 2001 at 14 897 patients, was interrupted in 2002 by the implementation of the allocation system based on the model for end stage liver disease and pediatric end-stage liver disease (MELD/PELD). Activity in all areas of intestinal transplantation continues to increase. One-year patient and graft survival following intestine-alone transplantation now seem to be superior to outcomes following liver-intestine transplantation. Other topics covered here include the recent 'Share 15' component of the MELD allocation system; liver transplantation following donation after cardiac death; simultaneous liver-kidney transplantation and waiting list and post-transplant outcomes for both liver and intestine transplantation, broken out by a variety of clinical and demographic factors. PMID- 17428287 TI - Heart and lung transplantation in the United States, 1996-2005. AB - This article examines the Organ Procurement and Transplantation Network/Scientific Registry of Transplant Recipients data on heart and lung transplantation in the United States from 1996 to 2005. The number of heart transplants performed and the size of the heart waiting list continued to drop, reaching 2126 and 1334, respectively, in 2005. Over the decade, post-transplant graft and patient survival improved, as did the chances for survival while on the heart waiting list. The number of deceased donor lung transplants increased by 78% since 1996, reaching 1407 in 2005 (up 22% from 2004). There were 3170 registrants awaiting lung transplantation at the end of 2005, down 18% from 2004. Death rates for both candidates and recipients have been dropping, as has the time spent waiting for a lung transplant. Other lung topics covered are living donation, recent surgical advances and changes in immunosuppression regimens. Heart-lung transplantation has declined to a small (33 procedures in 2005) but important need in the United States. PMID- 17428288 TI - Developing organ offer and acceptance measures: when 'good' organs are turned down. AB - Turndowns of offers of deceased donor kidneys for transplantation can contribute to inefficiencies in the organ distribution system and inequality in access to donated organs. Match run data were obtained for 4967 'good' kidneys placed and transplanted in 2005 after fewer than 50 offers. These kidneys were not recovered from donation after cardiac death or expanded criteria donors, or from donors with a history of substance abuse. On average, these good kidneys were not accepted until after seven offers to candidates and after offers to 2.4 programs. Models for the likelihood of acceptance found several donor and candidate characteristics to be significantly related to acceptance rates (p < 0.05). After accounting for these variables, there remained 2- to 3-fold differences among transplant programs in acceptance rates. These models could be used to identify kidney transplant centers with exceptional acceptance practices. Several strategies might be employed to increase acceptance rates for good organs. PMID- 17428289 TI - Geographic variability in access to primary kidney transplantation in the United States, 1996-2005. AB - This article focuses on geographic variability in patient access to kidney transplantation in the United States. It examines geographic differences and trends in access rates to kidney transplantation, in the component rates of wait listing, and of living and deceased donor transplantation. Using data from Centers for Medicare and Medicaid Services and the Organ Procurement and Transplantation Network/Scientific Registry of Transplant Recipients, we studied 700,000+ patients under 75, who began chronic dialysis treatment, received their first living donor kidney transplant, or were placed on the waiting list pre emptively. Relative rates of wait-listing and transplantation by State were calculated using Cox regression models, adjusted for patient demographics. There were geographic differences in access to the kidney waiting list and to a kidney transplant. Adjusted wait-list rates ranged from 37% lower to 64% higher than the national average. The living donor rate ranged from 57% lower to 166% higher, while the deceased donor transplant rate ranged from 60% lower to 150% higher than the national average. In general, States with higher wait-listing rates tended to have lower transplantation rates and States with lower wait-listing rates had higher transplant rates. Six States demonstrated both high wait-listing and deceased donor transplantation rates while six others, plus D.C. and Puerto Rico, were below the national average for both parameters. PMID- 17428290 TI - Repeat organ transplantation in the United States, 1996-2005. AB - The prospect of graft loss is a problem faced by all transplant recipients, and retransplantation is often an option when loss occurs. To assess current trends in retransplantation, we analyzed data for retransplant candidates and recipients over the last 10 years, as well as current outcomes. During 2005, retransplant candidates represented 13.5%, 7.9%, 4.1% and 5.5% of all newly registered kidney, liver, heart and lung candidates, respectively. At the end of 2005, candidates for retransplantation accounted for 15.3% of kidney transplant candidates, and lower proportions of liver (5.1%), heart (5.3%) and lung (3.3%) candidates. Retransplants represented 12.4% of kidney, 9.0% of liver, 4.7% of heart and 5.3% of lung transplants performed in 2005. The absolute number of retransplants has grown most notably in kidney transplantation, increasing 40% over the last 10 years; the relative growth of retransplantation was most marked in heart and lung transplantation, increasing 66% and 217%, respectively. The growth of liver retransplantation was only 11%. Unadjusted graft survival remains significantly lower after retransplantation in the most recent cohorts analyzed. Even with careful case mix adjustments, the risk of graft failure following retransplantation is significantly higher than that observed for primary transplants. PMID- 17428291 TI - Role of myocardial contractility and autonomic control in the hypotensive response to a limited access ethanol paradigm in SHRs. AB - BACKGROUND: Previous experimental studies that evaluated the chronic hemodynamic effect of ethanol employed the continuous exposure protocol of ethanol, which does not mimic the pattern of alcohol consumption in humans. This study dealt with the long-term hemodynamic and cardiovascular autonomic effects of ethanol, in a limited-access regimen in telemetered spontaneously hypertensive rats (SHRs). METHODS: Changes in blood pressure (BP), heart rate (HR), myocardial contractility (dP/dt(max)), and spectral cardiovascular autonomic profiles during the ethanol exposure period (2.5 or 5% w/v, 8 h/d, 8:30 am till 4:30 pm) were followed for 12 weeks. RESULTS: Compared with control pair-fed SHRs, body weight and urine output, osmolality, and potassium levels were decreased in SHRs receiving 5% but not 2.5% ethanol. Blood pressure showed progressive falls during ethanol-feeding periods with a maximum effect observed at week 5. The peak hypotensive effect was maintained thereafter in SHRs receiving 5% ethanol in contrast to steady rises in BP in the 2.5% ethanol group to near-control levels by the conclusion of the study. Heart rate was slightly but significantly increased by ethanol 5% whereas dP/dt(max) showed persistent reductions. Power spectral analysis showed that ethanol attenuated the baroreflex gain of HR as suggested by the reductions in index alpha, the spectral index of spontaneous baroreflex sensitivity (BRS). CONCLUSIONS: It is concluded that limited access ethanol drinking in SHRs elicited hypotension that was concentration dependent and mediated, at least partly, through reductions in myocardial contractility. Baroreflex sensitivity attenuation by ethanol appeared to have limited the tachycardic response to ethanol and perhaps its capacity to offset the evoked hypotension. PMID- 17428292 TI - Long-term ethanol self-administration by the nonhuman primate, Macaca fascicularis, decreases the benzodiazepine sensitivity of amygdala GABA(A) receptors. AB - BACKGROUND: Rodent models of chronic alcohol exposure are typically constrained to relatively short periods of forced ethanol due to the lifespan of these animals. Nonhuman primate models, particularly those employing long-term self administration, are conceptually more similar to human alcoholic individuals. METHODS: We performed whole-cell patch clamp recordings on acutely dissociated amygdala neurons isolated from cynomolgus macaque coronal temporal lobe slices. Slices were prepared from control monkeys or monkeys allowed to self-administer oral ethanol for 18 months. Flunitrazepam and acute ethanol modulation of currents gated by exogenous gamma-aminobutyric acid (GABA) application was assessed in these isolated neurons. Complementary experiments were performed on amygdala total RNA using quantitative real-time reverse transcription/polymerase chain reaction to understand potential ethanol-dependent adaptations to subunit composition. RESULTS: Gamma-aminobutyric acid-gated currents from ethanol-exposed macaque amygdala neurons exhibited reduced modulation by flunitrazepam compared with control neurons. However, this was specific for benzodiazepines as the modest inhibition of GABA-gated currents by acute ethanol was not affected by the chronic ethanol consumption. We also measured mRNA expression levels for the beta, gamma, and delta subunits in total amygdala RNA isolated from control and ethanol-drinking animals. beta1 and gamma2 expression was significantly reduced in samples from ethanol-exposed amygdala. CONCLUSIONS: Our findings demonstrate that chronic ethanol self-administration reduces the benzodiazepine sensitivity of amygdala GABA(A) receptors. This reduced sensitivity may be the result of decreased expression of an amygdala gamma subunit. These findings suggest that, while rodent and nonhuman primate models of chronic ethanol exposure share many characteristics, the specific molecular adaptations associated with the amygdala GABAergic system may not be identical. PMID- 17428293 TI - L-Buthionine (S,R) sulfoximine depletes hepatic glutathione but protects against ethanol-induced liver injury. AB - BACKGROUND: L-Buthionine (S,R) sulfoximine (BSO) is an inhibitor of glutathione biosynthesis and has been used as an effective means of depleting glutathione from cells and tissues. Here we investigated whether treatment with BSO enhanced ethanol-induced liver injury in mice. METHODS: Female C57Bl/6 mice were pair fed with control and ethanol-containing liquid diets in which ethanol was 29.2% of total calories. During the final 7 days of pair feeding, groups of control-fed and ethanol-fed mice were given 0, 5 or 7.6 mM BSO in the liquid diets. RESULTS: Compared with controls, ethanol given alone decreased total liver glutathione. This effect was exacerbated in mice given ethanol with 7.6 mM BSO, causing a 72% decline in hepatic glutathione. While ethanol alone caused no decrease in mitochondrial glutathione, inclusion of 7.6 mM BSO caused a 2-fold decline compared with untreated controls. L-Buthionine (S,R) sulfoximine did not affect ethanol consumption, but serum ethanol levels in BSO-treated mice were nearly 6 fold lower than in mice given ethanol alone. The latter decline in serum ethanol was associated with a significant elevation in the specific activities of cytochrome P450 2E1 and alcohol dehydrogenase in livers of BSO-treated animals. Ethanol consumption caused a 3.5-fold elevation in serum alanine aminotransferase levels but the enzyme fell to control levels when BSO was included in the diet. L Buthionine (S,R) sulfoximine administration also attenuated ethanol-induced steatosis, prevented the leakage of lysosomal cathepsins into the cytosol, and prevented the ethanol-elicited decline in proteasome activity. CONCLUSIONS: L Buthionine (S,R) sulfoximine, administered with ethanol, significantly depleted hepatic glutathione, compared with controls. However, despite the decrease in hepatic antioxidant levels, liver injury by ethanol was alleviated, due, in part, to a BSO-elicited acceleration of ethanol metabolism. PMID- 17428294 TI - Serum free sialic acid as a marker of alcohol abuse. AB - BACKGROUND: Previous studies have shown that serum total sialic acid (TSA) concentration significantly increases during alcohol abuse. Chronic ethanol consumption impairs glycosylation of many proteins. The increased desialylation rate of serum glycoproteins is one of the effects of alcohol abuse. The aim of this study was to investigate the diagnostic value of free sialic acid (FSA) as a marker of alcohol abuse. METHODS: We determined serum FSA concentrations in the group of 156 alcoholic subjects and 35 healthy control subjects by means of a modification of the thiobarbituric acid method. The alcoholic group was divided into subgroups according to their history of abuse. RESULTS: The FSA concentration was significantly higher in alcoholic subjects than in healthy controls. The subjects who consumed alcohol for longer than a week showed significantly higher FSA level than those who consumed alcohol for a shorter period. The serum FSA concentration was significantly higher in alcoholic subjects with elevated markers of liver dysfunction. The diagnostic accuracy of FSA was high, although it did not differ from TSA, and was limited by its low sensitivity. CONCLUSIONS: This study shows that FSA concentration in the sera of alcoholic subjects is increased. The low diagnostic sensitivity is accompanied by high specificity, however the accuracy is high and similar to the accuracy of TSA. Free sialic acid does not seem to be a better marker of alcohol abuse than TSA and current markers. PMID- 17428295 TI - Influence of subject eligibility criteria on compliance with National Institutes of Health guidelines for inclusion of women, minorities, and children in treatment research. AB - BACKGROUND: Many alcohol treatment outcome studies exclude some patients with particular problems, such as psychiatric disorders, noncompliance, and homelessness. Such criteria may increase the likelihood of a study being successfully conducted, but may also have the unintended consequence of reducing a study's ability to comply with National Institutes of Health guidelines for inclusion of racial minorities, women, and children in treatment research. METHODS AND RESULTS: This paper examined this issue empirically using 5 prior studies of treatment systems enrolling over 100,000 alcohol patients. Widely used eligibility criteria in the alcohol treatment field typically exclude between one fifth to one-third of patients from enrolling in research. Under several eligibility criteria, most notably those for drug use and social/residential instability, women and African-American patients are substantially more likely to be excluded than are men and non-African-American patients, respectively. CONCLUSIONS: In designing treatment studies with many eligibility criteria, researchers may therefore inadvertently be thwarting their own good faith efforts to ensure that a range of vulnerable populations are able to participate in research. We analyze the implications of this dilemma for the generalizability of treatment results and for research design, and provide data that may help researchers working in different treatment systems estimate the impact of various eligibility criteria. PMID- 17428296 TI - Heterogeneous effects of alcohol on dopamine release in the striatum: a PET study. AB - BACKGROUND: A dopaminergic response to alcohol in humans has not been demonstrated consistently with positron emission tomography (PET). We hypothesized that the effect of alcohol on striatal dopamine (DA) release may be anatomically heterogeneous between subjects. Our approach was to identify voxels that exhibited alcohol-induced DA responses within the striatum, and to determine the relationships between DA responses and alcohol-related behavior. METHODS: A novel method was developed to examine the anatomic extent and magnitude of striatal DA responses to alcohol across subjects. Thirteen healthy control subjects underwent 2 PET scans with [11C]raclopride (1 at baseline, 1 with an i.v. alcohol infusion to a target breath alcohol concentration of either 60 or 80 mg%). Parametric images of striatal binding potential (BP) were used to create maps of change in BP (deltaBP, an index of changes in DA levels). The anatomic extent and magnitude of DA responses were determined with voxel extraction methods. Subjective responses ("High," "Intoxication") to the alcohol infusion and behavioral data from the 90-day time-line follow back were assessed for relationships with DA responses to alcohol. RESULTS: A voxel-wise t-test between baseline and alcohol BP images did not show any differences in D2/D3 receptor availability between the conditions. Data from the striatal deltaBP maps nevertheless showed that the anatomic extent and magnitude of alcohol-induced DA release in the striatum are correlated with subjective responses to alcohol. CONCLUSIONS: The heterogeneity of dopaminergic responses to alcohol across subjects may be a reason for the lack of reports demonstrating DA involvement in alcohol-related behaviors. By allowing for different spatial patterns of DA release within each subject's striata, we showed correlations between alcohol induced DA release in the striata and behavioral outcomes related to alcohol. PMID- 17428297 TI - Alcohol effects on behavioral control: the impact of likelihood and magnitude of negative consequences. AB - BACKGROUND: This study utilized Signal Detection Theory to examine the relative influence of likelihood and magnitude of consequences when assessing alcohol effects on behavioral control. METHODS: Moderate to heavy drinkers (N=101) were assigned to alcohol or placebo and completed 4 administrations of a go/stop task that varied the likelihood and magnitude of punishment. RESULTS: Participants in the alcohol condition correctly responded to fewer go signals, and showed a trend toward reduction in sensitivity to task stimuli. Alcohol effects on sensitivity to go and stop signals were most evident when the likelihood of rewards and punishments were similar whereas the magnitude of punishment was not related to the strength of alcohol effects. CONCLUSION: Results suggest that likelihood of punishment is a more powerful determinant of alcohol-induced disinhibition than is magnitude. PMID- 17428298 TI - Prevalence of trigeminal autonomic symptoms in migraine: a population-based study. AB - Epidemiological data on trigeminal unilateral autonomic symptoms in patients with migraine are scarce. The authors wanted to provide a population-based evaluation of the prevalence of unilateral autonomic features in migraine patients and an assessment of the expression of unilaterality of autonomic symptoms and head pain in patients with UAs compared to other migraine patients. A population based sample of 6000 inhabitants of the city of Essen in Germany was screened using a previously validated standard questionnaire. Three thousand three hundred and sixty subjects (56% of a total 6000) responded. 841 subjects had migraine, out of which 226 reported accompanying unilatral auetonomic symptoms (26.9%, CI 95% [23.9-30%]). Unilateral autonomic symptoms in patients with migraine are common and have been widely underestimated in the past. One out of four migraine patients regularly experiences one or more unilateral autonomic symptoms during their attack. Migraine patients with accompanying autonomic symptoms seem to experience their pain more unilateral and more severe than non-UA patients. PMID- 17428299 TI - A multicentre, double-blind, randomized, placebo-controlled, parallel group study of multiple treatments of botulinum toxin type A (BoNTA) for the prophylaxis of episodic migraine headaches. AB - Our aim was to evaluate the safety and efficacy of botulinum toxin type A (BoNTA; BOTOX) for prophylaxis of episodic migraine. In this double-blind, placebo controlled study, patients were randomized to 225, 150 or 75 U of BoNTA or placebo after a 30-day placebo run-in for three 90-day treatment cycles. The primary efficacy end-point was the mean reduction from baseline in the frequency of migraine episodes at day 180 in the placebo non-responder stratum. All groups (N = 495) improved, with no significant differences. At day 180, the frequency of migraine episodes was reduced from baseline means of 4.3, 4.7, 4.7 and 4.4 by 1.6, 1.7, 1.5 and 1.4 for BoNTA 225 U, 150 U and 75 U and placebo, respectively. The primary end-point was not met. Treatment-related adverse events were transient and mild to moderate. BoNTA treatment was safe and well tolerated but did not result in significantly greater improvement than placebo in this study. Several factors may have confounded the results. PMID- 17428300 TI - Laser-evoked cortical potentials in cluster headache. AB - Craniofacial nociceptive processing in patients (n = 25) suffering from unilateral cluster headache was assessed by laser-evoked cortical potentials (LEPs). Latencies and amplitudes of late (N2, P2) and middle-latency (N1) LEPs were measured in chronic (CCH, n = 9) and episodic cluster headache (ECH, n = 17). In CCH patients on headache side N1c occurred later and P2 amplitude was smaller than on the healthy control side. In active periods of ECH patients P2 latency was shorter on the headache side. In remission periods of ECH patients the N2P2 ratio was lower on the headache side. In 19 out of 26 examinations in 25 headache patients LEP deviated from normative data in healthy controls (n = 10) without any specific pattern of altered parameters. LEPs document pathological changes in craniofacial nociception in cluster headache. However, there seems to be no pathognomonic deviation pattern that enables reliable diagnosis of cluster headache and application of LEPs in further studies of pathophysiological mechanisms. PMID- 17428301 TI - Biogeochemistry of methane and methanogenic archaea in permafrost. AB - This study summarizes the findings of our research on the genesis of methane, its content and distribution in permafrost horizons of different age and origin. Supported by reliable data from a broad geographical sweep, these findings confirm the presence of methane in permanently frozen fine-grained sediments. In contrast to the omnipresence of carbon dioxide in permafrost, methane-containing horizons (up to 40.0 mL kg(-1)) alternate with strata free of methane. Discrete methane-containing horizons representing over tens of thousands of years are indicative of the absence of methane diffusion through the frozen layers. Along with the isotopic composition of CH(4) carbon (delta(13)C -64 per thousand to -99 per thousand), this confirms its biological origin and points to in situ formation of this biogenic gas. Using (14)C-labeled substrates, the possibility of methane formation within permafrost was experimentally shown, as confirmed by delta(13)C values. Extremely low values (near -99 per thousand) indicate that the process of CH(4) formation is accompanied by the substantial fractionation of carbon isotopes. For the first time, cultures of methane-forming archaea, Methanosarcina mazei strain JL01 VKM B-2370, Methanobacterium sp. strain M2 VKM B 2371 and Methanobacterium sp. strain MK4 VKM B-2440 from permafrost, were isolated and described. PMID- 17428302 TI - Separation of bacteria of the Clostridium leptum subgroup from the human colonic microbiota by fluorescence-activated cell sorting or group-specific PCR using 16S rRNA gene oligonucleotides. AB - Molecular methods based on 16S rRNA gene sequence analyses have shown that bacteria of the Clostridium leptum subgroup are predominant in the colonic microbiota of healthy humans; this subgroup includes bacteria that produce butyrate, a source of energy for intestinal epithelial cells. To improve our understanding of the species within this important group, separation methods using fluorescence-activated cell sorting (FACS) and specific PCR were combined with 16S rRNA gene sequence analyses. FACS was developed for bacteria labelled in situ with two rRNA oligonucleotide probes, namely EUB338-FITC for total bacteria and Clep866-CY5/cp or Fprau645-CY5 for bacteria of the C. leptum subgroup. Bacterial cell sorting allowed a selective recovery of members of the C. leptum subgroup from the human microbiota with efficiencies as high as 95%. Group specific PCR amplification of the C. leptum subgroup was developed, and temporal thermal gradient gel electrophoresis showed host-specific profiles with low complexity, with a sharing of common bands between individuals and bands stable over 2 months for the same individual. A library of 16S rRNA gene cloned sequences (106 sequences) was prepared with DNA obtained from both separation methods, and 15 distinct phylotypes were identified, among which 10 have no cultivable or currently cultivated representative in reference collections. PMID- 17428303 TI - Stress response of methanogenic archaea from Siberian permafrost compared with methanogens from nonpermafrost habitats. AB - We examined the survival potential of methanogenic archaea exposed to different environmental stress conditions such as low temperature (down to -78.5 degrees C), high salinity (up to 6 M NaCl), starvation (up to 3 months), long-term freezing (up to 2 years), desiccation (up to 25 days) and oxygen exposure (up to 72 h). The experiments were conducted with methanogenic archaea from Siberian permafrost and were complemented by experiments on well-studied methanogens from nonpermafrost habitats. Our results indicate a high survival potential of a methanogenic archaeon from Siberian permafrost when exposed to the extreme conditions tested. In contrast, these stress conditions were lethal for methanogenic archaea isolated from nonpermafrost habitats. A better adaptation to stress was observed at a low temperature (4 degrees C) compared with a higher one (28 degrees C). Given the unique metabolism of methanogenic archaea in general and the long-term survival and high tolerance to extreme conditions of the methanogens investigated in this study, methanogenic archaea from permafrost should be considered as primary candidates for possible subsurface Martian life. PMID- 17428304 TI - Changes in the diversity of pig ileal lactobacilli around weaning determined by means of 16S rRNA gene amplification and denaturing gradient gel electrophoresis. AB - Our study aimed to provide a comprehensive characterization of changes in porcine intestinal Lactobacillus populations around the time of weaning based on 16S rRNA gene amplification and denaturing gradient gel electrophoresis (DGGE). DNA was extracted from the ileal contents of piglets at weaning (28 days of age) and after 1, 2, 5 and 11 days. PCR amplicons (V2-V3 fragments of 16S rRNA genes) were separated using DGGE. Predominant bands were excised and sequenced after reamplification. A band corresponding to Lactobacillus salivarius was present 1 and 2 days post-weaning (pw), while Lactobacillus crispatus was detected only 1 and 11 days pw. Lactobacillus sobrius gave the most dominant band in all animals. The number of bands decreased from 13+/-3 at weaning to 9+/-1 at 5 days pw, but the species richness had recovered by 11 days pw. The similarity of profiles between sampling days was high for 1 and 2 days pw (>91%), but was low for 5 and 11 days pw (<59%). The diversity of the profiles was lower 5 days pw, based on the Shannon diversity index (0.83+/-0.076 vs. 1.02+/-0.127 at weaning, P=0.042), but had recovered to preweaning values by 11 days pw. The application of group specific DGGE showed that the Lactobacillus community within the porcine ileum undergoes dramatic, partly reversible changes as a consequence of weaning. PMID- 17428305 TI - The role of neutrophils and monocytic cells in controlling the initiation of Clostridium perfringens gas gangrene. AB - Clostridium perfringens is a common cause of the fatal disease gas gangrene (myonecrosis). Established gas gangrene is notable for a profound absence of neutrophils and monocytic cells (phagocytes), and it has been suggested that the bactericidal activities of these cells play an insignificant role in controlling the progression of the infection. However, large inocula of bacteria are needed to establish an infection in experimental animals, suggesting phagocytes may play a role in inhibiting the initiation of gangrene. Examination of tissue sections of mice infected with a lethal (1 x 10(9)) or sublethal (1 x 10(6)) inoculum of C. perfringens revealed that phagocyte infiltration in the first 3 h postinfection was inhibited with a lethal dose but not with a sublethal dose, indicating that exclusion of phagocytes begins very early in the infection cycle. Experiments in which mice were depleted of either circulating monocytes or neutrophils before infection with C. perfringens showed that monocytes play a role in inhibiting the onset of gas gangrene at intermediate inocula but, although neutrophils can slow the onset of the infection, they are not protective. These results suggest that treatments designed to increase monocyte infiltration and activate macrophages may lead to increased resistance to the initiation of gas gangrene. PMID- 17428306 TI - Use of a novel coinfection system reveals a role for Rac1, H-Ras, and CrkII phosphorylation in Helicobacter pylori-induced host cell actin cytoskeletal rearrangements. AB - The Helicobacter pylori CagA protein induces profound morphological changes in the host cytoskeleton and cell scattering, but the signalling involved is poorly understood. Pseudomonas aeruginosa also affects host actin cytoskeleton in a variety of ways by injecting the ExoS and ExoT toxins which encode N-terminal GTPase activating protein and C-terminal ADP-ribosyltransferase (ADPRT) activities. In this study we developed a novel coinfection assay to gain new insights into CagA effector protein functions. We found that P. aeruginosa injecting either ExoT or ExoS efficiently prevented the H. pylori-induced scattering phenotype. Both the Rho-GAP and the ADPRT domains of ExoS were needed to block the H. pylori-induced actin cytoskeletal rearrangements, whereas either domain of ExoT was sufficient for this activity. This strategy revealed common pathways subverted by different pathogens, and aided in the definition of signalling cascades that control the CagA-mediated cell scattering and elongation. We identified Crk adapter proteins, Rac1 and H-Ras, but not RhoA or Cdc42, which are the ExoS and/or ExoT targets, as crucial components of the CagA induced phenotype. In addition, we show that ADP-ribosylation of CrkII by ExoT blocks phosphorylation of CrkII at Y-221, which is also important for the CagA induced signalling. PMID- 17428307 TI - Pathogenic and opportunistic free-living amoebae: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri, and Sappinia diploidea. AB - Among the many genera of free-living amoebae that exist in nature, members of only four genera have an association with human disease: Acanthamoeba spp., Balamuthia mandrillaris, Naegleria fowleri and Sappinia diploidea. Acanthamoeba spp. and B. mandrillaris are opportunistic pathogens causing infections of the central nervous system, lungs, sinuses and skin, mostly in immunocompromised humans. Balamuthia is also associated with disease in immunocompetent children, and Acanthamoeba spp. cause a sight-threatening infection, Acanthamoeba keratitis, mostly in contact-lens wearers. Of more than 30 species of Naegleria, only one species, N. fowleri, causes an acute and fulminating meningoencephalitis in immunocompetent children and young adults. In addition to human infections, Acanthamoeba, Balamuthia and Naegleria can cause central nervous system infections in animals. Because only one human case of encephalitis caused by Sappinia diploidea is known, generalizations about the organism as an agent of disease are premature. In this review we summarize what is known of these free living amoebae, focusing on their biology, ecology, types of disease and diagnostic methods. We also discuss the clinical profiles, mechanisms of pathogenesis, pathophysiology, immunology, antimicrobial sensitivity and molecular characteristics of these amoebae. PMID- 17428308 TI - Glucose controls multiple processes in Saccharomyces cerevisiae through diverse combinations of signaling pathways. AB - We have studied how the lack of glucose sensors in the plasma membrane, or of the enzymes Hxk1, Hxk2, Glk1, which catalyze the first intracellular step in glucose metabolism, affect the different responses of Saccharomyces cerevisiae to glucose. Lack of the G-protein-coupled receptor Gpr1 or of Snf3/Rgt2 did not affect glucose repression of different genes or activation by glucose of plasma membrane ATPase, whereas lack of Gpr1 decreased, in an additive manner with lack of Mth1, the degradation of fructose 1,6-bisphosphatase that takes place in the presence of glucose. In an hxk1 hxk2 glk1 strain, unable to phosphorylate glucose, all of these responses to the sugar were suppressed or strongly reduced. In the absence of Hxk2 (or Hxk1 and Hxk2), glucose repression of SUC2, GAL1 and GDH2 was relieved, but that of FBP1 and ICL1 was maintained. Hxk1 or Hxk2 were needed for activation of plasma membrane ATPase but not for degradation of FbPase. PMID- 17428309 TI - KNQ1, a Kluyveromyces lactis gene encoding a transmembrane protein, may be involved in iron homeostasis. AB - The original purpose of the experiments described in this article was to identify, in the biotechnologically important yeast Kluyveromyces lactis, gene(s) that are potentially involved in oxidative protein folding within the endoplasmic reticulum (ER), which often represents a bottleneck for heterologous protein production. Because treatment with the membrane-permeable reducing agent dithiothreitol inhibits disulfide bond formation and mimics the reducing effect that the normal transit of folding proteins has in the ER environment, the strategy was to search for genes that conferred higher levels of resistance to dithiothreitol when present in multiple copies. We identified a gene (KNQ1) encoding a drug efflux permease for several toxic compounds that in multiple copies conferred increased dithiothreitol resistance. However, the KNQ1 product is not involved in the excretion of dithiothreitol or in recombinant protein secretion. We generated a knq1 null mutant, and showed that both overexpression and deletion of the KNQ1 gene resulted in increased resistance to dithiothreitol. KNQ1 amplification and deletion resulted in enhanced transcription of iron transport genes, suggesting, for the membrane-associated protein Knq1p, a new, unexpected role in iron homeostasis on which dithiothreitol tolerance may depend. PMID- 17428310 TI - CD45-associated protein promotes the response of primary CD4 T cells to low potency T-cell receptor (TCR) stimulation and facilitates CD45 association with CD3/TCR and lck. AB - Although it is clear that the CD45 tyrosine phosphatase is required for efficient T-cell activation and T-cell development, the factors that regulate CD45 function remain uncertain. Previous data have indicated that there is an association of CD45 with CD4 and the T-cell receptor (TCR) complex controlled by the variable ectodomain of CD45 and, following activation, by high- and low-potency peptides. This suggests that controlling substrate access to CD45 may be an important regulatory mechanism during T-cell activation. In the present study we have examined the role of the transmembrane adapter-like molecule CD45-associated protein (CD45-AP) in regulating the association of CD45 with CD3/TCR and lck, and in regulating primary CD4(+) T-lymphocyte activation. In CD4(+) T cells from CD45 AP-deficient mice, coimmunoprecipitation of CD45 with the CD3/TCR complex, in addition to lck, is significantly reduced compared with wild-type T cells. Functionally, this correlates with a decreased proliferative response, a decrease in interleukin (IL)-2 production, and a decrease in calcium flux upon stimulation with a low-potency altered peptide ligand. However, the response of CD45-AP deficient T cells to stimulation with a high-avidity agonist peptide was largely intact, except for a modest decrease in IL-2 production. These data suggest that CD45-AP promotes or stabilizes the association of CD45 with substrates and regulates the threshold of T-cell activation. PMID- 17428311 TI - Sphingosine kinase inhibitor suppresses dendritic cell migration by regulating chemokine receptor expression and impairing p38 mitogen-activated protein kinase. AB - The migration of dendritic cells (DCs) to secondary lymphoid organs plays a crucial role in the initiation of adaptive immune responses. Although lipopolysaccharide enhances chemokine receptor 7 (CCR7) expression on DCs, the second signal for the migration of DCs toward the chemokine CCL19 remains unknown. In this study, we show that sphingosine kinase inhibitor (SKI) inhibits the migration of DCs toward CCL19 through the down-regulation of CCR7. Inhibition of p38 mitogen-activated protein kinase (MAPK) activation by SKI may be responsible for the SKI-mediated effects on the regulation of chemokine receptor expression. Impairment of DC migration by the inhibition of p38 MAPK and down regulation of CCR7 expression may contribute to the protective effects of SKI in DC-related disorders. These results suggest that sphingosine kinase-mediated signalling plays a role in the innate and adaptive immune responses by altering DC migration. PMID- 17428312 TI - Tolerogenic dendritic cells pulsed with enterobacterial extract suppress development of colitis in the severe combined immunodeficiency transfer model. AB - Immunomodulatory dendritic cells (DCs) that induce antigen-specific T-cell tolerance upon in vivo adoptive transfer are promising candidates for immunotherapy of autoimmune diseases. The feasibility of such a strategy has recently proved its efficacy in animal models of allotransplantation and experimental allergic encephalitis, but the effect in inflammatory bowel disease has not yet been demonstrated. In severe combined immunodeficient (SCID) mice, adoptively transferred CD4(+) CD25(-) T cells repopulate the lymphoid tissues and lead to development of chronic colitis characterized by CD4(+) T-cell proliferation against enterobacterial extract in vitro. In this model, we adoptively transferred in-vitro-generated bone-marrow-derived DCs exposed to interleukin-10 (IL-10) and an enterobacterial extract. We show that these cells are CD11c positive with intermediate expression of CD40, CD80 and CD86 and have a diminished secretion of IL-6, IL-12 p40/70, tumour necrosis factor-alpha and keratinocyte-derived chemokine (KC) compared to DCs treated with enterobacterial extract alone. In vivo, these cells prevented weight loss in SCID mice adoptively transferred with CD4(+) CD25(-) T cells, resulted in a lower histopathology colitis score and tended to result in higher serum levels of IL-1alpha, IL-10, IL 12, IL-13, IL-17, KC and monokine induced by interferon-gamma (MIG). These data underscore the potential of using immunomodulatory DCs to control inflammatory bowel disease and demonstrate its potential use in future human therapeutic settings. PMID- 17428313 TI - Immunological research using RNA interference technology. AB - RNA interference (RNAi) is a potent method of gene silencing that has developed rapidly over the past few years as a result of its extensive importance in the study of genetics, molecular biology and physiology. RNAi technology has also recently yielded significant insight into the innate and adaptive immune systems by helping to elucidate numerous mechanisms that regulate the development, activation and function of cells that mediate immunity. In addition, because of its ability to suppress gene expression effectively, this technique may be used to regulate the immune response for clinical purposes. Nonetheless, before RNAi can be successfully administered into human patients as a medical treatment, it is necessary to overcome several major limitations of this technology, such as inefficient in vivo delivery, incomplete silencing of target genes, non-specific immune responses, and off-target effects. As novel developments and discoveries in molecular biology swiftly continue to unfold, it is likely that RNAi may soon translate into a potent form of in vivo gene silencing with profound applications to vaccination and immunotherapy. In the present review, we examine the current progress of immunological studies employing RNAi and discuss the prospects for the implementation of this technique in the clinical arena. PMID- 17428314 TI - Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer. AB - INTRODUCTION: The impact of interactions between the two estrogen receptor (ER) subtypes, ERalpha and ERbeta, on gene expression in breast cancer biology is not clear. The goal of this study was to examine transcriptomic alterations in cancer cells co-expressing both receptors and the association of gene expression signatures with disease outcome. METHODS: Transcriptional effects of ERbeta overexpression were determined in a stably transfected cell line derived from ERalpha-positive T-47D cells. Microarray analysis was carried out to identify differential gene expression in the cell line, and expression of key genes was validated by quantitative polymerase chain reaction. Microarray and clinical data from patient samples were then assessed to determine the in vivo relevance of the expression profiles observed in the cell line. RESULTS: A subset of 14 DNA replication and cell cycle-related genes was found to be specifically downregulated by ERbeta. Expression profiles of four genes, CDC2, CDC6, CKS2, and DNA2L, were significantly inversely correlated with ERbeta transcript levels in patient samples, consistent with in vitro observations. Kaplan-Meier analysis revealed better disease outcome for the patient group with an expression signature linked to higher ERbeta expression as compared to the lower ERbeta expressing group for both disease-free survival (p = 0.00165) and disease specific survival (p = 0.0268). These findings were further validated in an independent cohort. CONCLUSION: Our findings revealed a transcriptionally regulated mechanism for the previously described growth inhibitory effects of ERbeta in ERalpha-positive breast tumor cells and provide evidence for a functional and beneficial impact of ERbeta in primary breast tumors. PMID- 17428315 TI - Constructing osteoarthritis through discourse--a qualitative analysis of six patient information leaflets on osteoarthritis. AB - BACKGROUND: Health service policy in the United Kingdom emphasises the importance of self-care by patients with chronic conditions. Written information for patients about their condition is seen as an important aid to help patients look after themselves. From a discourse analysis perspective written texts such as patient information leaflets do not simply describe the reality of a medical condition and its management but by drawing on some sorts of knowledge and evidence rather than others help construct the reality of that condition. This study explored patient information leaflets on osteoarthritis (OA) to see how OA was constructed and to consider the implications for self-care. METHODS: Systematic and repeated readings of six patient information leaflets on osteoarthritis to look for similarities and differences across leaflets, contradictions within leaflets and the resources called on to make claims about the nature of OA and its management. RESULTS: Biomedical discourse of OA as a joint disease dominated. Only one leaflet included an illness discourse albeit limited, and was also the only one to feature patient experiences of living with OA. The leaflets had different views on the causes of OA including the role of lifestyle and ageing. Most emphasised patient responsibility for preventing the progression of OA. Advice about changing behaviour such as diet and exercise was not grounded in lived experience. There were inconsistent messages about using painkillers, exercise and the need to involve professionals when making changes to lifestyle. CONCLUSION: The nature of the discourse impacted on how OA and the respective roles of patients and professionals were depicted. Limited discourse on illness meant that the complexity of living with OA and its consequences was underestimated. Written information needs to shift from joint biology to helping patients live with osteoarthritis. Written information should incorporate patient experience and value it alongside biomedical knowledge. PMID- 17428316 TI - Ovarian dysfunction and FMR1 alleles in a large Italian family with POF and FRAXA disorders: case report. AB - BACKGROUND: The association between premature ovarian failure (POF) and the FMR1 repeat number (41> CGGn< 200) has been widely investigated. Current findings suggest that the risk estimation for POF can be calculated in the offspring of women with pre-mutated FMR1 alleles. CASE PRESENTATION: We describe the coexistence in a large Italian kindred of Fragile X syndrome and familial POF in females with ovarian dysfunctions who carried normal or expanded FMR1 alleles. Genetic analysis of the FMR1 gene in over three generations of females revealed that six carried pre-mutated alleles (61-200), of which two were also affected by POF. However a young woman, who presented a severe ovarian failure with early onset, carried normal FMR1 alleles (<40). The coexistence within the same family of two dysfunctional ovarian conditions, one FMR1-related and one not FMR1 related, suggests that the complexity of familial POF conditions is larger than expected. CONCLUSION: Our case study represents a helpful observation and will provide familial cases with heterogeneous etiology that could be further studied when candidate genes in addition to the FMR1 premutation will be available. PMID- 17428317 TI - A previously unobserved conformation for the human Pex5p receptor suggests roles for intrinsic flexibility and rigid domain motions in ligand binding. AB - BACKGROUND: The C-terminal tetratricopeptide (TPR) repeat domain of Pex5p recognises proteins carrying a peroxisomal targeting signal type 1 (PTS1) tripeptide in their C-terminus. Previously, structural data have been obtained from the TPR domain of Pex5p in both the liganded and unliganded states, indicating a conformational change taking place upon cargo protein binding. Such a conformational change would be expected to play a major role both during PTS1 protein recognition as well as in cargo release into the peroxisomal lumen. However, little information is available on the factors that may regulate such structural changes. RESULTS: We have used a range of biophysical and computational methods to further analyse the conformational flexibility and ligand binding of Pex5p. A new crystal form for the human Pex5p C-terminal domain (Pex5p(C)) was obtained in the presence of Sr2+ ions, and the structure presents a novel conformation, distinct from all previous liganded and apo crystal structures for Pex5p(C). The difference relates to a near-rigid body movement of two halves of the molecule, and this movement is different from that required to reach a ring-like conformation upon PTS1 ligand binding. The bound Sr2+ ion changes the dynamic properties of Pex5p(C) affecting its conformation, possibly by making the Sr2+-binding loop - located near the hinge region for the observed domain motions - more rigid. CONCLUSION: The current data indicate that Pex5p(C) is able to sample a range of conformational states in the absence of bound PTS1 ligand. The domain movements between various apo conformations are distinct from those involved in ligand binding, although the differences between all observed conformations so far can be characterised by the movement of the two halves of Pex5p(C) as near-rigid bodies with respect to each other. PMID- 17428318 TI - Patient Engagement and Coaching for Health: The PEACH study--a cluster randomised controlled trial using the telephone to coach people with type 2 diabetes to engage with their GPs to improve diabetes care: a study protocol. AB - BACKGROUND: The PEACH study is based on an innovative 'telephone coaching' program that has been used effectively in a post cardiac event trial. This intervention will be tested in a General Practice setting in a pragmatic trial using existing Practice Nurses (PN) as coaches for people with type 2 diabetes (T2D). Actual clinical care often fails to achieve standards, that are based on evidence that self-management interventions (educational and psychological) and intensive pharmacotherapy improve diabetes control. Telephone coaching in our study focuses on both. This paper describes our study protocol, which aims to test whether goal focused telephone coaching in T2D can improve diabetes control and reduce the treatment gap between guideline based standards and actual clinical practice. METHODS/DESIGN: In a cluster randomised controlled trial, general practices employing Practice Nurses (PNs) are randomly allocated to an intervention or control group. We aim to recruit 546 patients with poorly controlled T2D (HbA1c >7.5%) from 42 General Practices that employ PNs in Melbourne, Australia. PNs from General Practices allocated to the intervention group will be trained in diabetes telephone coaching focusing on biochemical targets addressing both patient self-management and engaging patients to work with their General Practitioners (GPs) to intensify pharmacological treatment according to the study clinical protocol. Patients of intervention group practices will receive 8 telephone coaching sessions and one face-to-face coaching session from existing PNs over 18 months plus usual care and outcomes will be compared to the control group, who will only receive only usual care from their GPs. The primary outcome is HbA1c levels and secondary outcomes include cardiovascular disease risk factors, behavioral risk factors and process of care measures. DISCUSSION: Understanding how to achieve comprehensive treatment of T2D in a General Practice setting is the focus of the PEACH study. This study explores the potential role for PNs to help reduce the treatment and outcomes gap in people with T2D by using telephone coaching. The intervention, if found to be effective, has potential to be sustained and embedded within real world General Practice. PMID- 17428319 TI - Adjuvant TACE inhibitor treatment improves the outcome of TLR2-/- mice with experimental pneumococcal meningitis. AB - BACKGROUND: Streptococcus (S.) pneumoniae meningitis has a high lethality despite antibiotic treatment. Inflammation is a major pathogenetic factor, which is unresponsive to antibiotics. Therefore adjunctive therapies with antiinflammatory compounds have been developed. TNF484 is a TNF-alpha converting enzyme (TACE) inhibitor and has been found efficacious in experimental meningitis. Toll-like receptor 2 (TLR2) contributes to host response in pneumococcal meningitis by enhancing bacterial clearing and downmodulating inflammation. In this study, TNF484 was applied in mice, which lacked TLR2 and exhibited a strong meningeal inflammation. METHODS: 103 CFU S. pneumoniae serotype 3 was inoculated subarachnoidally into C57BL/6 wild type (wt) mice or TLR2-/-, CD14-/- and CD14-/ /TLR2-/- mice. Severity of disease and survival was followed over 9 days. Response to antibiotics (80 mg/kg ceftriaxone i.p. for 5 days) and/or TACE inhibitor treatment (1 mg/kg s.c. twice daily for 4 days) was evaluated. Animals were sacrificed after 12, 24, and 48 h for analysis of bacterial load in cerebrospinal fluid (CSF) and brain and for TNF and leukocyte measurements in CSF. RESULTS: TLR2-/- mice were significantly sicker than the other mouse strains 24 h after infection. All knockout mice showed higher disease severity after 48 h and died earlier than wt mice. TNF release into CSF was significantly more elevated in TLR2-/- than in the other strains after 24 h. Brain bacterial numbers were significantly higher in all knockout than wt mice after 24 h. Modulation of outcome by antibiotic and TACE inhibitor treatment was evaluated. With antibiotic therapy all wt, CD14-/- and TLR2-/-/CD14-/- mice, but only 79% of TLR2-/- mice, were rescued. TACE inhibitor treatment alone did not rescue, but prolonged survival in wt mice, and in TLR2-/- and CD14-/- mice to the values observed in untreated wt mice. By combined antibiotic and TACE inhibitor treatment 95% of TLR2-/- mice were rescued. CONCLUSION: During pneumococcal meningitis strong inflammation in TLR2-deficiency was associated with incomplete responsiveness to antibiotics and complete response to combined antibiotic and TACE inhibitor treatment. TACE inhibitor treatment offers a promising adjuvant therapeutic strategy in pneumococcal meningitis. PMID- 17428320 TI - Identification of women with an increased risk of developing radiation-induced breast cancer: a case only study. AB - INTRODUCTION: Radiation exposure at a young age is one of the strongest risk factors for breast cancer. Germline mutations in genes involved in the DNA-damage repair pathway (DDRP) may render women more susceptible to radiation-induced breast cancer. METHODS: We evaluated the contribution of germline mutations in the DDRP genes BRCA1, BRCA2, CHEK2 and ATM to the risk of radiation-induced contralateral breast cancer (CBC). The germline mutation frequency was assessed, in a case-only study, in women who developed a CBC after they had a first breast cancer diagnosed before the age of 50 years, and who were (n = 169) or were not (n = 78) treated with radiotherapy for their first breast tumour. RESULTS: We identified 27 BRCA1, 5 BRCA2, 15 CHEK2 and 4 truncating ATM germline mutation carriers among all CBC patients tested (21%). The mutation frequency was 24.3% among CBC patients with a history of radiotherapy, and 12.8% among patients not irradiated for the first breast tumour (odds ratio 2.18 (95% confidence interval 1.03 to 4.62); p = 0.043). The association between DDRP germline mutation carriers and risk of radiation-induced CBC seemed to be strongest in women who developed their second primary breast tumour at least 5 years after radiotherapy. Those patients had an odds ratio of 2.51 (95% confidence interval 1.03 to 6.10; p = 0.049) of developing radiation-induced breast cancer, in comparison with non carriers. CONCLUSION: This study shows that carriers of germline mutations in a DDRP gene have an increased risk of developing (contralateral) breast cancer after radiotherapy; that is, over and above the risk associated with their carrier status. The increased risk indicates that knowledge of germline status of these DDRP genes at the time of breast cancer diagnosis may have important implications for the choice of treatment. PMID- 17428321 TI - Comparative genomics and experimental promoter analysis reveal functional liver specific elements in mammalian hepatic lipase genes. AB - BACKGROUND: Mammalian hepatic lipase (HL) genes are transcribed almost exclusively in hepatocytes. The basis for this liver-restricted expression is not completely understood. We hypothesized that the responsible cis-acting elements are conserved among mammalian HL genes. To identify these elements, we made a genomic comparison of 30 kb of 5'-flanking region of the rat, mouse, rhesus monkey, and human HL genes. The in silico data were verified by promoter-reporter assays in transfected hepatoma HepG2 and non-hepatoma HeLa cells using serial 5' deletions of the rat HL (-2287/+9) and human HL (-685/+13) promoter region. RESULTS: Highly conserved elements were present at the proximal promoter region, and at 14 and 22 kb upstream of the transcriptional start site. Both of these upstream elements increased transcriptional activity of the human HL (-685/+13) promoter region 2-3 fold. Within the proximal HL promoter region, conserved clusters of transcription factor binding sites (TFBS) were identified at -240/ 200 (module A), -80/-40 (module B), and -25/+5 (module C) by the rVista software. In HepG2 cells, modules B and C, but not module A, were important for basal transcription. Module B contains putative binding sites for hepatocyte nuclear factors HNF1alpha. In the presence of module B, transcription from the minimal HL promoter was increased 1.5-2 fold in HepG2 cells, but inhibited 2-4 fold in HeLa cells. CONCLUSION: Our data demonstrate that searching for conserved non-coding sequences by comparative genomics is a valuable tool in identifying candidate enhancer elements. With this approach, we found two putative enhancer elements in the far upstream region of the HL gene. In addition, we obtained evidence that the -80/-40 region of the HL gene is responsible for enhanced HL promoter activity in hepatoma cells, and for silencing HL promoter activity in non-liver cells. PMID- 17428322 TI - Diagnosis of left ventricular diastolic dysfunction in the setting of acute changes in loading conditions. AB - INTRODUCTION: Conventional pulsed wave Doppler parameters are known to be preload dependent, whereas newly proposed Doppler indices may be less influenced by variations in loading conditions. The aim of the present study was to evaluate the effects of haemodialysis-induced preload reduction on both conventional and new Doppler parameters for the assessment of left ventricular (LV) diastolic function. METHODS: This prospective observational study was conducted in a medical-surgical intensive care unit (ICU) and nephrology department of a teaching hospital. In total, 37 haemodialysis patients with end-stage renal disease (age [mean +/- standard deviation]: 52 +/- 13 years) and eight ventilated ICU patients with acute renal failure receiving vasopressor therapy (age 57 +/- 16 years; Simplified Acute Physiology Score II 51 +/- 17) were studied. Echocardiography was performed before and after haemodialysis. Conventional pulsed wave Doppler indices of LV diastolic function as well as new Doppler indices, including Doppler tissue imaging early diastolic velocities (E' wave) of the septal and lateral portions of the mitral annulus, and propagation velocity of LV inflow at early diastole (Vp) were measured and compared before and after ultrafiltration. RESULTS: The volume of ultrafiltration was greater in haemodialysis patients than in ICU patients (3.0 +/- 1.1 l versus 1.9 +/- 0.9 l; P = 0.005). All conventional pulsed wave Doppler parameters were altered by haemodialysis. In haemodialysis patients, E' velocity decreased after ultrafiltration when measured at the septal mitral annulus (7.1 +/- 2.5 cm/s versus 5.9 +/- 1.7 cm/s; P = 0.0003), but not at its lateral portion (8.9 +/- 3.1 cm/s versus 8.3 +/- 2.6 cm/s; P = 0.37), whereas no significant variation was observed in ICU patients. Vp decreased uniformly after ultrafiltration, the difference being significant only in haemodialysis patients (45 +/- 11 cm/s versus 41 +/- 13 cm/s; P = 0.04). Although of less magnitude, ultrafiltration induced variations in Doppler parameters were also observed in haemodialysis patients with altered LV systolic function. CONCLUSION: In contrast to other Doppler parameters, Doppler tissue imaging E' maximal velocity measured at the lateral mitral annulus represents an index of LV diastolic function that is relatively insensitive to abrupt and marked preload reduction. PMID- 17428323 TI - RepSeq--a database of amino acid repeats present in lower eukaryotic pathogens. AB - BACKGROUND: Amino acid repeat-containing proteins have a broad range of functions and their identification is of relevance to many experimental biologists. In human-infective protozoan parasites (such as the Kinetoplastid and Plasmodium species), they are implicated in immune evasion and have been shown to influence virulence and pathogenicity. RepSeq http://repseq.gugbe.com is a new database of amino acid repeat-containing proteins found in lower eukaryotic pathogens. The RepSeq database is accessed via a web-based application which also provides links to related online tools and databases for further analyses. RESULTS: The RepSeq algorithm typically identifies more than 98% of repeat-containing proteins and is capable of identifying both perfect and mismatch repeats. The proportion of proteins that contain repeat elements varies greatly between different families and even species (3-35% of the total protein content). The most common motif type is the Sequence Repeat Region (SRR)--a repeated motif containing multiple different amino acid types. Proteins containing Single Amino Acid Repeats (SAARs) and Di-Peptide Repeats (DPRs) typically account for 0.5-1.0% of the total protein number. Notable exceptions are P. falciparum and D. discoideum, in which 33.67% and 34.28% respectively of the predicted proteomes consist of repeat-containing proteins. These numbers are due to large insertions of low complexity single and multi-codon repeat regions. CONCLUSION: The RepSeq database provides a repository for repeat-containing proteins found in parasitic protozoa. The database allows for both individual and cross-species proteome analyses and also allows users to upload sequences of interest for analysis by the RepSeq algorithm. Identification of repeat-containing proteins provides researchers with a defined subset of proteins which can be analysed by expression profiling and functional characterisation, thereby facilitating study of pathogenicity and virulence factors in the parasitic protozoa. While primarily designed for kinetoplastid work, the RepSeq algorithm and database retain full functionality when used to analyse other species. PMID- 17428324 TI - Spatial and temporal patterns of enzootic raccoon rabies adjusted for multiple covariates. AB - BACKGROUND: With the objective of identifying spatial and temporal patterns of enzootic raccoon variant rabies, a spatial scan statistic was utilized to search for significant terrestrial rabies clusters by year in New York State in 1997 2003. Cluster analyses were unadjusted for other factors, adjusted for covariates, and adjusted for covariates and large scale geographic variation (LSGV). Adjustments were intended to identify the unusual aggregations of cases given the expected distribution based on the observed locations. RESULTS: Statistically significant clusters were identified particularly in the Albany, Finger Lakes, and South Hudson areas. The clusters were generally persistent in the Albany area, but demonstrated cyclical changes in rabies activity every few years in the other areas. Cluster adjustments allowed the discussion of possible causes for the high risk raccoon rabies areas identified. CONCLUSION: This study analyzed raccoon variant rabies spatial and temporal patterns in New York that have not been previously described at a focal (census tract) level. Comparisons across the type of spatial analysis performed with various degrees of adjustment allow consideration of the potential influence of geographical factors for raccoon rabies and possible reasons for the highest risk areas (statistically significant clusters). PMID- 17428325 TI - Common variants in the ATM, BRCA1, BRCA2, CHEK2 and TP53 cancer susceptibility genes are unlikely to increase breast cancer risk. AB - INTRODUCTION: Certain rare, familial mutations in the ATM, BRCA1, BRCA2, CHEK2 or TP53 genes increase susceptibility to breast cancer but it has not, until now, been clear whether common polymorphic variants in the same genes also increase risk. METHODS: We have attempted a comprehensive, single nucleotide polymorphism (SNP)- and haplotype-tagging association study on each of these five genes in up to 4,474 breast cancer cases from the British, East Anglian SEARCH study and 4,560 controls from the EPIC-Norfolk study, using a two-stage study design. Nine tag SNPs were genotyped in ATM, together with five in BRCA1, sixteen in BRCA2, ten in CHEK2 and five in TP53, with the aim of tagging all other known, common variants. SNPs generating the common amino acid substitutions were specifically forced into the tagging set for each gene. RESULTS: No significant breast cancer associations were detected with any individual or combination of tag SNPs. CONCLUSION: It is unlikely that there are any other common variants in these genes conferring measurably increased risks of breast cancer in our study population. PMID- 17428326 TI - Geographic prediction of tuberculosis clusters in Fukuoka, Japan, using the space time scan statistic. AB - BACKGROUND: Tuberculosis (TB) has reemerged as a global public health epidemic in recent years. Although evaluating local disease clusters leads to effective prevention and control of TB, there are few, if any, spatiotemporal comparisons for epidemic diseases. METHODS: TB cases among residents in Fukuoka Prefecture between 1999 and 2004 (n = 9,119) were geocoded at the census tract level (n = 109) based on residence at the time of diagnosis. The spatial and space-time scan statistics were then used to identify clusters of census tracts with elevated proportions of TB cases. RESULTS: In the purely spatial analyses, the most likely clusters were in the Chikuho coal mining area (in 1999, 2002, 2003, 2004), the Kita-Kyushu industrial area (in 2000), and the Fukuoka urban area (in 2001). In the space-time analysis, the most likely cluster was the Kita-Kyushu industrial area (in 2000). The north part of Fukuoka Prefecture was the most likely to have a cluster with a significantly high occurrence of TB. CONCLUSION: The spatial and space-time scan statistics are effective ways of describing circular disease clusters. Since, in reality, infectious diseases might form other cluster types, the effectiveness of the method may be limited under actual practice. The sophistication of the analytical methodology, however, is a topic for future study. PMID- 17428327 TI - Data Mining in Bioinformatics (BIOKDD). PMID- 17428328 TI - Decisions by regulatory agencies: are they evidence-based? AB - Contradictory statements about the non-steroidal anti-inflammatory drugs from the European Medicines Agency and the United States Food and Drug Administration have raised questions about whether regulatory decisions are evidence-based. For the selective COX-2 inhibitors, there are clear contraindications and warnings in Europe, but only a vaguely worded Black Box warning in the United States. All the non-selective agents are given an almost "clean bill of health" in Europe, while all of them are judged to have a similar risk-benefit ratio as celecoxib in the United States. The regulatory agencies have failed to recognize the clinical trial evidence that the risk of cardiovascular events varies substantially among the non-selective agents, with diclofenac carrying the highest risk of harm. PMID- 17428329 TI - Comparative genomics using Fugu reveals insights into regulatory subfunctionalization. AB - BACKGROUND: A major mechanism for the preservation of gene duplicates in the genome is thought to be mediated via loss or modification of cis-regulatory subfunctions between paralogs following duplication (a process known as regulatory subfunctionalization). Despite a number of gene expression studies that support this mechanism, no comprehensive analysis of regulatory subfunctionalization has been undertaken at the level of the distal cis regulatory modules involved. We have exploited fish-mammal genomic alignments to identify and compare more than 800 conserved non-coding elements (CNEs) that associate with genes that have undergone fish-specific duplication and retention. RESULTS: Using the abundance of duplicated genes within the Fugu genome, we selected seven pairs of teleost-specific paralogs involved in early vertebrate development, each containing clusters of CNEs in their vicinity. CNEs present around each Fugu duplicated gene were identified using multiple alignments of orthologous regions between single-copy mammalian orthologs (representing the ancestral locus) and each fish duplicated region in turn. Comparative analysis reveals a pattern of element retention and loss between paralogs indicative of subfunctionalization, the extent of which differs between duplicate pairs. In addition to complete loss of specific regulatory elements, a number of CNEs have been retained in both regions but may be responsible for more subtle levels of subfunctionalization through sequence divergence. CONCLUSION: Comparative analysis of conserved elements between duplicated genes provides a powerful approach for studying regulatory subfunctionalization at the level of the regulatory elements involved. PMID- 17428330 TI - Statistical tools for synthesizing lists of differentially expressed features in related experiments. AB - We propose a novel approach for finding a list of features that are commonly perturbed in two or more experiments, quantifying the evidence of dependence between the experiments by a ratio. We present a Bayesian analysis of this ratio, which leads us to suggest two rules for choosing a cut-off on the ranked list of p values. We evaluate and compare the performance of these statistical tools in a simulation study, and show their usefulness on two real datasets. PMID- 17428331 TI - A major zebrafish polymorphism resource for genetic mapping. AB - We have identified 645,088 candidate polymorphisms in zebrafish and observe a single nucleotide polymorphism (SNP) validation rate of 71% to 86%, improving with polymorphism confidence score. Variant sites are non-random, with an excess of specific novel T- and A-rich motifs. We positioned half of the polymorphisms on zebrafish genetic and physical maps as a resource for positional cloning. We further demonstrate bulked segregant analysis using the anchored SNPs as a method for high-throughput genetic mapping in zebrafish. PMID- 17428332 TI - Global transcriptome analysis of murine embryonic stem cell-derived cardiomyocytes. AB - BACKGROUND: Characterization of gene expression signatures for cardiomyocytes derived from embryonic stem cells will help to define their early biologic processes. RESULTS: A transgenic alpha-myosin heavy chain (MHC) embryonic stem cell lineage was generated, exhibiting puromycin resistance and expressing enhanced green fluorescent protein (EGFP) under the control of the alpha-MHC promoter. A puromycin-resistant, EGFP-positive, alpha-MHC-positive cardiomyocyte population was isolated with over 92% purity. RNA was isolated after electrophysiological characterization of the cardiomyocytes. Comprehensive transcriptome analysis of alpha-MHC-positive cardiomyocytes in comparison with undifferentiated alpha-MHC embryonic stem cells and the control population from 15-day-old embryoid bodies led to identification of 884 upregulated probe sets and 951 downregulated probe sets in alpha-MHC-positive cardiomyocytes. A subset of upregulated genes encodes cytoskeletal and voltage-dependent channel proteins, and proteins that participate in aerobic energy metabolism. Interestingly, mitosis, apoptosis, and Wnt signaling-associated genes were downregulated in the cardiomyocytes. In contrast, annotations for genes upregulated in the alpha-MHC positive cardiomyocytes are enriched for the following Gene Ontology (GO) categories: enzyme-linked receptor protein signaling pathway (GO:0007167), protein kinase activity (GO:0004672), negative regulation of Wnt receptor signaling pathway (GO:0030178), and regulation of cell size (O:0008361). They were also enriched for the Biocarta p38 mitogen-activated protein kinase signaling pathway and Kyoto Encyclopedia of Genes and Genomes (KEGG) calcium signaling pathway. CONCLUSION: The specific pattern of gene expression in the cardiomyocytes derived from embryonic stem cells reflects the biologic, physiologic, and functional processes that take place in mature cardiomyocytes. Identification of cardiomyocyte-specific gene expression patterns and signaling pathways will contribute toward elucidating their roles in intact cardiac function. PMID- 17428333 TI - Core strength: a new model for injury prediction and prevention. AB - OBJECTIVE: Many work in injury prone awkward positions that require adequate flexibility and strength in trunk stabilizer muscle groups. Performance on a functional movement screen (FMS) that assessed those factors was conducted and an intervention was designed. METHODS: A battery of FMS tests were performed on 433 firefighters. We analyzed the correlation between FMS performance and injuries and other selected parameters. An intervention to improve flexibility and strength in trunk stabilizer or core muscle groups through a training program was evaluated. RESULTS: The intervention reduced lost time due to injuries by 62% and the number of injuries by 42% over a twelve month period as compared to a historical control group. CONCLUSION: These findings suggest that core strength and functional movement enhancement programs to prevent injuries in workers whose work involves awkward positions is warranted. PMID- 17428334 TI - Rapid detection of Pfcrt and Pfmdr1 mutations in Plasmodium falciparum isolates by FRET and in vivo response to chloroquine among children from Osogbo, Nigeria. AB - BACKGROUND: Chloroquine (CQ) has been in use in Africa for a long time. Because of misuse, this drug has now lost its efficacy due to the emergence of resistance strains in most parts of Africa. Recently, it was shown that after chloroquine has been withdrawn from the market, chloroquine-sensitive Plasmodium falciparum re-emerged and chloroquine could again be used successfully as an antimalarial. Surveillance of parasite populations is, therefore, important to decide whether chloroquine could be re-introduced. METHODS: To estimate the prevalence of the most pivotal polymorphisms, including Pfcrt K76T, Pfmdr1 N86Y and Pfmdr1 Y184F mutations, and their contributions to the outcome of CQ treatment, isolates from Osogbo Western Nigeria were tested using the Fluorescence Resonance Energy Transfer (FRET) method on a real-time PCR instrument. RESULTS: 116 children with acute uncomplicated P. falciparum malaria infections were treated with the standard dosage of CQ and followed-up for 28 days. Blood samples were collected on filter paper at enrollment and during follow-up for identification of parasite carrying the chloroquine resistant transporter (pfcrt) and P. falciparum-multi drug resistance (pfmdr1) gene mutations. Parasitological assessment of response to treatment showed that 62% of the patients were cured and 38% failed the CQ treatment. The presence of single mutant pfcrt (T76) alleles (P = 0.003) and in combination with mutant pfmdr1 Y86 (P = 0.028) was significantly associated with in vivo CQR. No other mutation on its own or in combinations was significantly associated with treatment outcome. Mutant pfcrt was more prevalent in both pre- and post-treatment isolates. No association was observed between age or initial level of parasitaemia and chloroquine treatment outcome. CONCLUSION: The result established the usefulness and accuracy of real time PCR in pfcrt and pfmdr1 mutation detection and also give further evidence to the reliability of the pfcrt T76 point mutation as a molecular marker for CQ resistance. PMID- 17428335 TI - Identification of a robust gene signature that predicts breast cancer outcome in independent data sets. AB - BACKGROUND: Breast cancer is a heterogeneous disease, presenting with a wide range of histologic, clinical, and genetic features. Microarray technology has shown promise in predicting outcome in these patients. METHODS: We profiled 162 breast tumors using expression microarrays to stratify tumors based on gene expression. A subset of 55 tumors with extensive follow-up was used to identify gene sets that predicted outcome. The predictive gene set was further tested in previously published data sets. RESULTS: We used different statistical methods to identify three gene sets associated with disease free survival. A fourth gene set, consisting of 21 genes in common to all three sets, also had the ability to predict patient outcome. To validate the predictive utility of this derived gene set, it was tested in two published data sets from other groups. This gene set resulted in significant separation of patients on the basis of survival in these data sets, correctly predicting outcome in 62-65% of patients. By comparing outcome prediction within subgroups based on ER status, grade, and nodal status, we found that our gene set was most effective in predicting outcome in ER positive and node negative tumors. CONCLUSION: This robust gene selection with extensive validation has identified a predictive gene set that may have clinical utility for outcome prediction in breast cancer patients. PMID- 17428336 TI - Curative and organ-preserving treatment with intra-arterial carboplatin induction followed by surgery and/or radiotherapy for advanced head and neck cancer: single center five-year results. AB - BACKGROUND: This study evaluated the feasibility, toxicity, response rate and survival of neoadjuvant superselective intra-arterial infusion of high dose carboplatin in advanced head and neck cancer. METHODS: Forty-six patients with primary head and neck squamous cell carcinoma received 3 cycles of intra-arterial carboplatin (300 to 350 mg/m2 per cycle every 2 weeks), followed by radiotherapy or surgery plus radiotherapy. RESULTS: No complications or severe toxicity occurred. Sixteen patients (35%) were complete responders, 20 (43%) partial responders while 10 (22%) did not respond to treatment. After completion of the multimodality treatment, 38/46 patients (83%) were complete responders. After a 5 year follow-up period, 18/46 patients (39%) are alive and disease-free, 3 (6,5%) have died of a second primary tumor and 25 (54.5%) have died of the disease. CONCLUSION: Intra-arterial carboplatin induction chemotherapy is a safe, well tolerated technique that discriminates between responders and non-responders and so may have prognostic significance in planning further integrated treatments aimed to organ preservation for advanced head and neck carcinomas. PMID- 17428337 TI - Coronary artery-pulmonary artery fistula: case report. AB - BACKGROUND: Coronary artery fistulas are rare congenital or acquired coronary artery anomalies that can originate from any of the three major coronary arteries and drain in all the cardiac chambers and great vessels. CASE PRESENTATION: An 11 year-old boy was referred for evaluation of an exertional dyspnoea. He reported recent history of few episodes of shortness of breath associated with moderate entity physical activity. At physical examination a mild continuous murmur could be heard mainly at the level of the second intercostal space of the left parasternal area. A transthoracic echocardiogram showed a continuous flow at color Doppler analysis in the high parasternal short axis view, originating from a small entry site on the wall of the main pulmonary artery. A selective left coronary angiography revealed a fistula connecting the proximal portion of the left anterior descending coronary artery with the main pulmonary artery. CONCLUSION: A combination like the one described in the present case is unusual since fistulas originate from the left coronary artery in about 35% of cases and drainage into the pulmonary artery occurs in only 17%. PMID- 17428338 TI - The concentration of oxygen, lactate and glucose in the central veins, right heart, and pulmonary artery: a study in patients with pulmonary hypertension. AB - INTRODUCTION: Decreases in oxygen saturation (SO2) and lactate concentration [Lac] from superior vena cava (SVC) to pulmonary artery have been reported. These gradients (Delta SO2 and Delta[Lac]) are probably created by diluting SVC blood with blood of lower SO2 and [Lac]. We tested the hypothesis that Delta SO2 and Delta[Lac] result from mixing SVC and inferior vena cava (IVC) blood streams. METHODS: This was a prospective, sequential, observational study of hemodynamically stable individuals with pulmonary artery hypertension (n = 9) who were about to undergo right heart catheterization. Catheters were advanced under fluoroscopic guidance into the IVC, SVC, right atrium, right ventricle, and pulmonary artery. Samples were obtained at each site and analyzed for SO2, [Lac], and glucose concentration ([Glu]). Analysis of variance with Tukey HSD test was used to compare metabolite concentrations at each site. RESULTS: There were no differences in SO2 or [Lac] between IVC and SVC, both being greater than their respective pulmonary artery measurements (P < 0.01 for SO2 and P < 0.05 for [Lac]). SO2 and [Lac] in right atrium, right ventricle, and pulmonary artery were similar. Delta SO2 was 4.4 +/- 1.4% (mean +/- standard deviation) and Delta[Lac] was 0.16 +/- 0.11 mmol/l (both > 0; P < 0.001). Delta[Glu] was -0.19 +/- 0.31 mmol/l, which was not significantly different from zero, with SVC [Glu] being less than IVC [Glu]. CONCLUSION: Mixing of SVC with IVC blood does not account for the development of Delta SO2 and Delta[Lac] in hemodynamically stable individuals with pulmonary artery hypertension. An alternate mechanism is mixing with coronary sinus blood, implying that Delta SO2 and Delta[Lac] may reflect changes in coronary sinus SO2 and [Lac] in this patient population. PMID- 17428339 TI - Discovery of chemically induced mutations in rice by TILLING. AB - BACKGROUND: Rice is both a food source for a majority of the world's population and an important model system. Available functional genomics resources include targeted insertion mutagenesis and transgenic tools. While these can be powerful, a non-transgenic, unbiased targeted mutagenesis method that can generate a range of allele types would add considerably to the analysis of the rice genome. TILLING (Targeting Induced Local Lesions in Genomes), a general reverse genetic technique that combines traditional mutagenesis with high throughput methods for mutation discovery, is such a method. RESULTS: To apply TILLING to rice, we developed two mutagenized rice populations. One population was developed by treatment with the chemical mutagen ethyl methanesulphonate (EMS), and the other with a combination of sodium azide plus methyl-nitrosourea (Az-MNU). To find induced mutations, target regions of 0.7-1.5 kilobases were PCR amplified using gene specific primers labeled with fluorescent dyes. Heteroduplexes were formed through denaturation and annealing of PCR products, mismatches digested with a crude preparation of CEL I nuclease and cleaved fragments visualized using denaturing polyacrylamide gel electrophoresis. In 10 target genes screened, we identified 27 nucleotide changes in the EMS-treated population and 30 in the Az MNU population. CONCLUSION: We estimate that the density of induced mutations is two- to threefold higher than previously reported rice populations (about 1/300 kb). By comparison to other plants used in public TILLING services, we conclude that the populations described here would be suitable for use in a large scale TILLING project. PMID- 17428340 TI - Health related quality of life and psychosocial function among patients with carcinoid tumours. A longitudinal, prospective, and comparative study. AB - BACKGROUND: The aim was to investigate HRQoL and psychosocial function among patients with carcinoid tumours, longitudinally and prospectively, and to compare HRQoL among patients with carcinoid tumours to that of the Swedish general population. The aim was also to investigate the prevalence of distress during the first year after diagnosis. METHODS: At four assessments during the first year after diagnosis, HRQoL was measured by the EORTC QLQ-C30 3.0, anxiety and depression by the HADS, and prevalence, and worst aspects of distress by an interview guide. ANOVA was performed in order to study changes over time with regard to HRQoL, anxiety and depression. Comparisons regarding HRQoL between patients and the Swedish population were made by the use of one-sample t-tests and changes over time regarding the prevalence of distress was investigated by means of Cochran's Q. RESULTS: High levels of physical-, emotional-, cognitive-, and social function and somewhat lower levels of role function and global quality of life were reported at all assessments. Role- and emotional function increased over time. Patients reported lower role function and global quality of life and more problems with fatigue and diarrhoea than the Swedish general population, at all assessments. Fatigue, limitations to work and pursue daily activities, and worry that the illness will get worse were among the most prevalent aspects at all assessments. At all assessments the majority reported worrying about the family's situation, the ability to care for the family, and worrying before the check-up. CONCLUSION: It is concluded that HRQoL and psychosocial function among patients with carcinoid tumours remains stable during the first year, that the patients report a lower HRQoL than the Swedish general population, and that a majority of the patients report a number of aspects of emotional distress. In the clinical care, it should be considered that the majority of patients report not only fatigue and diarrhoea but also worries about their prognosis, their families, tests, and examinations. Efforts to reduce these worries should be made. PMID- 17428341 TI - How high is the level of technical noise in microarray data? AB - BACKGROUND: Microarray gene expression data are commonly perceived as being extremely noisy because of many imperfections inherent in the current technology. A recent study conducted by the MicroArray Quality Control (MAQC) Consortium and published in Nature Biotechnology provides a unique opportunity to probe into the true level of technical noise in such data. RESULTS: In the present report, the MAQC study is reanalyzed in order to quantitatively assess measurement errors inherent in high-density oligonucleotide array technology (Affymetrix platform). The level of noise is directly estimated from technical replicates of gene expression measurements in the absence of biological variability. For each probe set, the magnitude of random fluctuations across technical replicates is characterized by the standard deviation of the corresponding log-expression signal. The resultant standard deviations appear to be uniformly small and symmetrically distributed across probe sets. The observed noise level does not cause any tangible bias in estimated pair-wise correlation coefficients, the latter being particularly prone to its presence in microarray data. CONCLUSION: The reported analysis strongly suggests that, contrary to popular belief, the random fluctuations of gene expression signals caused by technical noise are quite low and the effect of such fluctuations on the results of statistical inference from Affymetrix GeneChip microarray data is negligibly small. PMID- 17428343 TI - The clinical effect of a new infant formula in term infants with constipation: a double-blind, randomized cross-over trial. AB - BACKGROUND: Nutrilon Omneo (new formula; NF) contains high concentration of sn-2 palmitic acid, a mixture of prebiotic oligosaccharides and partially hydrolyzed whey protein. It is hypothesized that NF positively affects stool characteristics in constipated infants. METHODS: Thirty-eight constipated infants, aged 3-20 weeks, were included and randomized to NF (n = 20) or a standard formula (SF; n = 18) in period 1 and crossed-over after 3 weeks to treatment period 2. Constipation was defined by at least one of the following symptoms: 1) defecation frequency < 3/week; 2) painful defecation; 3) abdominal or rectal palpable mass. RESULTS: Period 1 was completed by 35 infants. A significant increase in defecation frequency (NF: 3.5 pre versus 5.6/week post treatment; SF 3.6 pre versus 4.9/week post treatment) was found in both groups, but was not significantly different between the two formulas (p = 0.36). Improvement of hard stool consistency to soft stool consistency was found more often with NF than SF, but did not reach statistical significance (90% versus 50%; RR, 1.8; 95% CI, 0.9 3.5; p = 0.14). No difference was found in painful defecation or the presence of an abdominal or rectal mass between the two groups. Twenty-four infants completed period 2. Only stool consistency was significantly different between the two formulas (17% had soft stools on NF and hard stools on SF; no infants had soft stools on SF and hard stools on NF, McNemar test p = 0.046). CONCLUSION: The addition of a high concentration sn-2 palmitic acid, prebiotic oligosaccharides and partially hydrolyzed whey protein resulted in a strong tendency of softer stools in constipated infants, but not in a difference in defecation frequency. Formula transition to NF may be considered as treatment in constipated infants with hard stools. PMID- 17428344 TI - Flexible mapping of homology onto structure with homolmapper. AB - BACKGROUND: Over the past decade, a number of tools have emerged for the examination of homology relationships among protein sequences in a structural context. Most recent software implementations for such analysis are tied to specific molecular viewing programs, which can be problematic for collaborations involving multiple viewing environments. Incorporation into larger packages also adds complications for users interested in adding their own scoring schemes or in analyzing proteins incorporating unusual amino acid residues such as selenocysteine. RESULTS: We describe homolmapper, a command-line application for mapping information from a multiple protein sequence alignment onto a protein structure for analysis in the viewing software of the user's choice. Homolmapper is small (under 250 K for the application itself) and is written in Python to ensure portability. It is released for non-commercial use under a modified University of California BSD license. Homolmapper permits facile import of additional scoring schemes and can incorporate arbitrary additional amino acids to allow handling of residues such as selenocysteine or pyrrolysine. Homolmapper also provides tools for defining and analyzing subfamilies relative to a larger alignment, for mutual information analysis, and for rapidly visualizing the locations of mutations and multi-residue motifs. CONCLUSION: Homolmapper is a useful tool for analysis of homology relationships among proteins in a structural context. There is also extensive, example-driven documentation available. More information about homolmapper is available at http://www.mcb.ucdavis.edu/faculty labs/lagarias/homolmapper_home/homolmapper%20web%20page.htm. PMID- 17428345 TI - A phase I, randomized study of combined IL-2 and therapeutic immunisation with antiretroviral therapy. AB - BACKGROUND: Fully functional HIV-1-specific CD8 and CD4 effector T-cell responses are vital to the containment of viral activity and disease progression. These responses are lacking in HIV-1-infected patients with progressive disease. We attempted to augment fully functional HIV-1-specific CD8 and CD4 effector T-cell responses in patients with advanced chronic HIV-1 infection. DESIGN: Chronically infected patients with low CD4 counts T-cell counts who commenced antiretroviral therapy (ART) were subsequently treated with combined interleukin-2 and therapeutic vaccination. METHODS: Thirty six anti-retroviral naive patients were recruited and initiated on combination ART for 17 weeks before randomization to: A) ongoing ART alone; B) ART with IL-2 twice daily for 5 days every four weeks starting at week 17 for 3 cycles; C) ART with IL-2 as in group B and Remune HIV-1 vaccine administered once every 3 months, starting at week 17; and D) ART with Remune vaccine as in group C. Patients were studied for 65 weeks following commencement of ART, with an additional prior 6 week lead-in observation period. CD4 and CD8 T-cell counts, evaluations of HIV-1 RNA levels and proliferative responses to recall and HIV-1 antigens were complemented with assessment of IL-4 secretion alongside quantification of anti-HIV-1 CD8 T-cell responses and neutralizing antibody titres. RESULTS: Neither IL-2 nor Remune vaccination induced sustained HIV-1-specific T-cell responses. However, we report an inverse relationship between HIV-1-specific proliferative responses and IL-4 production which continuously increased in patients receiving immunotherapy, but not patients receiving ART alone. CONCLUSION: Induction of HIV-1-specific cell mediated responses is a major challenge in chronically HIV-1-infected patients even when combining immunisation with IL-2 therapy. An antigen-specific IL-4 associated suppressive response may play a role in attenuating HIV-specific responses. PMID- 17428342 TI - The use of global transcriptional analysis to reveal the biological and cellular events involved in distinct development phases of Trichophyton rubrum conidial germination. AB - BACKGROUND: Conidia are considered to be the primary cause of infections by Trichophyton rubrum. RESULTS: We have developed a cDNA microarray containing 10250 ESTs to monitor the transcriptional strategy of conidial germination. A total of 1561 genes that had their expression levels specially altered in the process were obtained and hierarchically clustered with respect to their expression profiles. By functional analysis, we provided a global view of an important biological system related to conidial germination, including characterization of the pattern of gene expression at sequential developmental phases, and changes of gene expression profiles corresponding to morphological transitions. We matched the EST sequences to GO terms in the Saccharomyces Genome Database (SGD). A number of homologues of Saccharomyces cerevisiae genes related to signalling pathways and some important cellular processes were found to be involved in T. rubrum germination. These genes and signalling pathways may play roles in distinct steps, such as activating conidial germination, maintenance of isotropic growth, establishment of cell polarity and morphological transitions. CONCLUSION: Our results may provide insights into molecular mechanisms of conidial germination at the cell level, and may enhance our understanding of regulation of gene expression related to the morphological construction of T. rubrum. PMID- 17428346 TI - Two novel missense mutations in the myostatin gene identified in Japanese patients with Duchenne muscular dystrophy. AB - BACKGROUND: Myostatin is a negative regulator of skeletal muscle growth. Truncating mutations in the myostatin gene have been reported to result in gross muscle hypertrophy. Duchenne muscular dystrophy (DMD), the most common lethal muscle wasting disease, is a result of an absence of muscle dystrophin. Although this disorder causes a rather uniform pattern of muscle wasting, afflicted patients display phenotypic variability. We hypothesized that genetic variation in myostatin is a modifier of the DMD phenotype. METHODS: We analyzed 102 Japanese DMD patients for mutations in the myostatin gene. RESULTS: Two polymorphisms that are commonly observed in Western countries, p.55A>T and p.153K>R, were not observed in these Japanese patients. An uncommon polymorphism of p.164E>K was uncovered in four cases; each patient was found to be heterozygous for this polymorphism, which had the highest frequency of the polymorphism observed in the Japanese patients. Remarkably, two patients were found to be heterozygous for one of two novel missense mutations (p.95D>H and p.156L>I). One DMD patient carrying a novel missense mutation of p.95D>H was not phenotypically different from the non-carriers. The other DMD patient was found to carry both a novel mutation (p.156L>I) and a known polymorphism (p.164E>K) in one allele, although his phenotype was not significantly modified. Any nucleotide change creating a target site for micro RNAs was not disclosed in the 3' untranslated region. CONCLUSION: Our results indicate that heterozygous missense mutations including two novel mutations did not produce an apparent increase in muscle strength in Japanese DMD cases, even in a patient carrying two missense mutations. PMID- 17428347 TI - Severe sepsis due to severe falciparum malaria and leptospirosis co-infection treated with activated protein C. AB - Co-infection with falciparum malaria and leptospirosis is uncommon. The aim of this study is to report a case of severe sepsis secondary to dual infection with falciparum malaria and leptospirosis. The literature is also reviewed on the clinical course of such co-infections, and the possible mechanisms and treatment of patients with life-threatening malaria and leptospirosis with activated protein C. The patient was a 25-year old male admitted in the Respiratory Intensive Care Unit (RICU) with fever, haemolysis, acute renal failure, hepatitis, acute lung injury (ALI) and altered sensorium. A syndromic evaluation was done and investigations revealed falciparum parasitaemia. He was treated with parenteral artesunate, ceftriaxone and doxycycline, and adjunctive therapies as for severe sepsis. Infusion of activated protein C was started 20 hours after onset of organ dysfunction, and intensive haemodialysis was instituted. Over the next four days the patient became afebrile with progressive resolution of ALI, renal failure and hepatitis. His Leptospira serology (requested as part of the evaluation) was reported positive on day 5. Dual infections are common and under recognized in the tropics. Failure to treat potential co-infections may lead to poor outcomes. Acute lung injury in falciparum malaria has high mortality rates and therapy as for severe sepsis may improve survival. Adjunctive therapies, including activated protein C, cannot replace source eradication. PMID- 17428348 TI - Alternative splicing variations in mouse CAPS2: differential expression and functional properties of splicing variants. AB - BACKGROUND: Ca2+-dependent activator protein 2 (CAPS2/CADPS2) is a secretory vesicle-associated protein involved in the release of neurotrophin. We recently reported that an aberrant, alternatively spliced CAPS2 mRNA that lacks exon 3 (CAPS2Deltaexon3) is detected in some patients with autism. Splicing variations in mouse CAPS2 and their expression and functions remain unclear. RESULTS: In this study, we defined 31 exons in the mouse CAPS2 gene and identified six alternative splicing variants, CAPS2a-f. CAPS2a is an isoform lacking exons 22 and 25, which encode part of the Munc13-1-homologous domain (MHD). CAPS2b lacks exon 25. CAPS2c lacks exons 11 and 22. CAPS2d, 2e, and 2f have C-terminal deletions from exon 14, exon 12, and exon 5, respectively. On the other hand, a mouse counterpart of CAPS2Deltaexon3 was not detected in the mouse tissues tested. CAPS2b was expressed exclusively in the brain, and the other isoforms were highly expressed in the brain, but also in some non-neural tissues. In the brain, all isoforms showed predominant expression patterns in the cerebellum. In the developing cerebellum, CAPS2b showed an up-regulated expression pattern, whereas the other isoforms exhibited transiently peaked expression patterns. CAPS2 proteins were mostly recovered in soluble fractions, but some were present in membrane fractions, except for CAPS2c and 2f, both of which lack the PH domain, suggesting that the PH domain is important for membrane association. In contrast to CAPS2a and 2b, CAPS2c showed slightly decreased BDNF-releasing activity, which is likely due to the C-terminal truncation of the PH domain in CAPS2c. CONCLUSION: This study indicates that, in mouse, there are six splicing variants of CAPS2 (CAPS2a-f), and that these are subdivided into two groups: a long form containing the C-terminal MHD and a short form lacking the C-terminal MHD. These results demonstrate that the splicing variations correlate with their expression patterns and intracellular distribution, and affect BDNF release; however, whether or not the short forms possess activities other than BDNF release, for example as natural dominant-negative isoforms, remains to be determined. PMID- 17428349 TI - Association of TGFbeta1, TNFalpha, CCR2 and CCR5 gene polymorphisms in type-2 diabetes and renal insufficiency among Asian Indians. AB - BACKGROUND: Cytokines play an important role in the development of diabetic chronic renal insufficiency (CRI). Transforming growth factor beta1 (TGF beta1) induces renal hypertrophy and fibrosis, and cytokines like tumor necrosis factor alpha (TNFalpha), chemoattractant protein-1 (MCP-1), and regulated upon activation and normal T cell expressed and secreted (RANTES) mediate macrophage infiltration into kidney. Over expression of these chemokines leads to glomerulosclerosis and interstitial fibrosis. The effect of MCP-1 and RANTES on kidney is conferred by their receptors i.e., chemokine receptor (CCR)-2 and CCR-5 respectively. We tested association of nine single nucleotide polymorphisms (SNPs) from TGFbeta1, TNFalpha, CCR2 and CCR5 genes among individuals with type-2 diabetes with and without renal insufficiency. METHODS: Type-2 diabetes subjects with chronic renal insufficiency (serum creatinine > or = 3.0 mg/dl) constituted the cases, and matched individuals with diabetes of duration > or = 10 years and normoalbuminuria were evaluated as controls from four centres in India. Allelic and genotypic contributions of nine SNPs from TGFbeta1, TNFalpha, CCR2 and CCR5 genes to diabetic CRI were tested by computing odds ratio (OR) and 95% confidence intervals (CI). Sub-analysis of CRI cases diabetic retinopathy status as dependent variable and SNP genotypes as independent variable in a univariate logistic regression was also performed. RESULTS: SNPs Tyr81His and Thr263Ile in TGF beta1 gene were monomorphic, and Arg25Pro in TGF beta1 gene and Delta32 polymorphism in CCR5 gene were minor variants (minor allele frequency <0.05) and therefore were not considered for case-control analysis. A significant allelic association of 59029G>A SNP of CCR5 gene has been observed and the allele 59029A seems to confer predisposition to development of diabetic CRI (OR 1.39; CI 1.04 1.84). In CRI subjects a compound group of genotypes "GA and AA" of SNP G>A -800 was found to confer predisposition for proliferative retinopathy (OR 3.03; CI 1.08-8.50, p = 0.035). CONCLUSION: Of the various cytokine gene polymorphisms tested, allele 59029A of CCR5 gene is significantly associated with diabetic renal insufficiency among Asian Indians. Result obtained for 59029G>A SNP of CCR5 gene is in conformity with reports from a Japanese population but due to sub optimal power of the sample, replication in larger sample set is warranted. PMID- 17428350 TI - Monitoring and analysis of dynamic growth of human embryonic stem cells: comparison of automated instrumentation and conventional culturing methods. AB - BACKGROUND: Human embryonic stem cells (hESCs) are a potential source of cells for use in regenerative medicine. Automation of culturing, monitoring and analysis is crucial for fast and reliable optimization of hESC culturing methods. Continuous monitoring of living cell cultures can reveal more information and is faster than using laborious traditional methods such as microscopic evaluation, immunohistochemistry and flow cytometry. METHODS: We analyzed the growth dynamics of two hESC lines HS237 and HS293 in a conventional culture medium containing serum replacement and a xeno-free X-vivo 10 medium. We used a new automated culture platform utilizing machine vision technology, which enables automatic observation, recording and analysis of intact living cells. We validated the results using flow cytometry for cell counting and characterization. RESULTS: In our analyses, hESC colony growth could be continuously monitored and the proportion of undifferentiated cells automatically analyzed. No labeling was needed and we could, for the first time, perform detailed follow up of live, undisturbed cell colonies, and record all the events in the culture. The growth rate of the hESCs cultured in X-vivo 10 medium was significantly lower and a larger proportion of the cells were differentiated. CONCLUSION: The new automated system enables rapid and reliable analysis of undifferentiated growth dynamics of hESCs. We demonstrate the effectiveness of the system by comparing hESC growth in different culture conditions. PMID- 17428351 TI - Interspecific and intraspecific interactions in the monogenean communities of fish: a question of study scale? AB - Monogenean communities of fish have generally been considered non-interactive as negative interspecific interactions have rarely been reported. Most of the earlier studies on monogenean communities, however, have been conducted not only in systems with relatively low parasite abundances but, more importantly, at study scales where microhabitat-level interactions between the parasites are easily overlooked. We examined the communities of 3 abundant Dactylogyrus (Monogenea) species on the gills of crucian carp (Carassius carassius) by analysing the interactions at the scale of individual gill filaments, where interactions between the species, if any, should most likely take place. Contrary to our expectations, we did not find evidence for competitive exclusion between the species, which suggests that monogenean communities are non-interactive even in high parasite abundances. At the species level, individual parasites were highly aggregated within the filaments, essentially showing a strong tendency to occur at either end of a filament. This, together with the result of differences in the distribution of juvenile parasites within the filaments compared to adults, suggests that these parasites are able to actively seek out their conspecifics in small-scale microhabitats during maturation, which again could enhance their mate-finding. PMID- 17428352 TI - The anti-schistosomal drug praziquantel is an adenosine antagonist. AB - The mechanism of action of praziquantel (PZQ), the drug of choice against schistosomiasis, is still unclear. Since exposure of schistosomes to the drug is associated with calcium influx and muscular contraction, calcium channels have been suggested as the target, although direct combination of PZQ with their subunits was never demonstrated. We report a hitherto unknown effect of PZQ, namely the inhibition of nucleoside uptake, as observed in living worms using radio-isotope labelled adenosine and uridine. This effect is clearly seen in schistosomes but is absent in mammalian cells in culture. Moreover it is a specific pharmacological effect seen exclusively with the active levo-R(-)stereo isomer of the drug, and is shared by at least one benzodiazepine having antischistosomal activity. This novel effect acquires significance given that schistosomes cannot synthesize purine nucleosides de novo. A possible relationship between this novel effect and the known action of PZQ on calcium channels is discussed, since adenosine is known to bind to specific receptors and to behave as an indirect antagonist of calcium release in mammalian cells. If calcium channels were correlated with adenosine receptors also in schistosomes, as they are in mammals, this would support the hypothesis that PZQ-induced calcium influx may be correlated to adenosine receptor blockade. PMID- 17428353 TI - Evolutionary diversity in polystomatids infecting tetraploid and octoploid Xenopus in East African highlands: biological and molecular evidence. AB - Species of Protopolystoma are monogenean flukes that only infect allopolyploid hosts in the anuran genus Xenopus. Multivariate analyses of morphometric sclerite characters in the nominal species Protopolystoma simplicis suggest that morphologically distinguishable populations occur in the tetraploid host, Xenopus laevis victorianus, and in each of the octoploid hosts, X. vestitus and X. wittei. The species-level divergence of a lineage specific to X. laevis is supported by sequence variation in the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Protopolystoma simplicis from X. laevis is redesignated P. microsclera n. sp., with P. simplicis being retained for populations in octoploid hosts. This division is consistent with large differences in egg hatching schedule, fixed differences at the mannose-6-phosphate isomerase and fumarate hydratase loci, and host-specificity in experimental analyses. Although the respective P. simplicis populations in X. vestitus and X. wittei also show significant diversity in allozyme expression, morphometrics and egg hatching schedule, they are retained in the same species because their level of mitochondrial DNA divergence is similar to that found within other Protopolystoma species. The consequences of splitting P. simplicis for a recent interpretation of the origin of Protopolystoma faunas in octoploid Xenopus spp. is discussed. PMID- 17428354 TI - A novel high sensitivity ELISA for detection of antineutrophil cytoplasm antibodies against proteinase-3. AB - OBJECTIVE: Conventional direct enzyme-linked immunosorbent assays (ELISA) for the detection of anti-neutrophil cytoplasm antibodies (ANCA) often lack sensitivity because epitopes of the target antigen are hidden by binding to the ELISA plate. This study was designed to evaluate a novel ELISA method for detection of ANCA against proteinase-3 (PR3) for the diagnosis of Wegener's granulomatosis (WG) using PR3 presented in its native form. METHODS: Sera from four subgroups of patients with a diagnosis of WG (n=86), 80 healthy controls and 450 disease controls were tested for the presence of C-ANCA/PR3-ANCA by anchor ELISA, direct ELISA, capture ELISA, indirect immunofluorescence (IFT) and immunoblotting. RESULTS: In prospectively analysed consecutive patients, anchor ELISA showed the highest sensitivity for a diagnosis of WG of 96.0% (95% CI: 79.6-99.3), followed by IFT 92.0% (73.9-98.8), capture ELISA 72.0 (50.6-87.9) and direct ELISA 60.0 (38.7-78.8). Specificity was high for all methods and ranged from 98.5 (97.0 99.4) to 95.5% (97.9-99.8). Receiver operating characteristics curve analysis revealed that the overall diagnostic performance of the anchor ELISA was significantly superior compared to the direct ELISA and the capture ELISA in patients with generalized WG, and also compared to IFT and immunoblotting in patients with localised WG. CONCLUSION: Anchor ELISA is a novel highly sensitive and specific method for the detection of PR3-ANCA in patients with WG, which may replace the need for a combined analysis with IFT and ELISA in the future. PMID- 17428355 TI - Lack of association between interleukin-6 promoter polymorphism at position -174 and Henoch-Schonlein pur pura. AB - OBJECTIVE: To assess whether polymorphism of the interleukin (IL)-6 gene at the position -174 was implicated in the incidence of Henoch-Schonlein pur-pura (HSP). A further objective was to determine if any relationship existed with severe systemic complications of HSP, in particular with severe renal and gastrointestinal involvement. METHODS: Unselected patients from Northwest Spain with primary cutaneous vasculitis classified as HSP according to proposed criteria were studied. All patients included in the present study were required to have had at least 2 year's follow-up. Patients and controls were genotyped for a single biallelic (G/C) nucleotide polymorphism in the promoter region at the position -174 of the IL-6 gene by a polymerase reaction chain-restriction fragment length polymorphism (PCR-RFLP) method. RESULTS: Forty-six Caucasian HSP patients and 124 healthy matched controls were studied. No allele or genotype differences between the whole group of HSP and controls were observed. This was also the case when HSP patients were stratified by the presence of gastrointestinal complications, nephritis, and permanent renal involvement (renal sequelae). CONCLUSION: The polymorphism in IL-6 gene promoter (-174 G/C) does not appear to be a genetic risk factor for HSP in Northwest Spain. PMID- 17428356 TI - Kidney involvement in Takayasu arteritis. AB - OBJECTIVE: To evaluate whether the presence of glomerulonephritis is or is not associated with the extent of arterial wall inflammatory cell infiltrate in Takayasu arthritis (TA). METHODS: Retrospective chart and pathology review of large artery and kidney specimens of TA autopsy cases. Kidney specimens were classified, according to their histopathological findings, in those with specific glomerular entities and those with non-specific, ischemic and/or hypertensive, glomerular changes. A control group of autopsy kidney specimens was utilized for comparison. Morphometric analysis was used to assess the extent of the arterial inflammatory infiltrates; results were compared among the different groups with kidney lesions. RESULTS: We included 25 kidney specimens from 25 autopsies. Specific glomerular entities were present in 14 specimens; 10 (40%) were classified as diffuse mesangial proliferative glomerulonephritis (DMPG [Group A]), and 4 (16%) as other associated glomerulopathies (Group B). Non-specific changes were observed in 11 (44%) specimens (Group C). The arterial inflammatory infiltrate proportion was 9.4 % for group A, 1.4% for group B, and 2.7% for group C. Furthermore, a larger proportion of vascular inflammation was confirmed for group A when compared with the other groups (p<0.05). Group A patients were younger than those in groups B and C (p<0.005) and exhibited shorter disease duration. CONCLUSION: The presence of DMPG was associated with a larger extent of vascular inflammatory cell infiltrate, suggesting a relationship between both phenomena. PMID- 17428357 TI - The incidence of giant cell arteritis in Jerusalem over a 25-year period: annual and seasonal fluctuations. AB - OBJECTIVE: Giant-cell arteritis (GCA) incidence is reported to be rising. A cyclic pattern of annual incidence rates and seasonal variations were reported by several groups. However, such fluctuations were not observed by others. We examined both annual and seasonal rates of GCA over a period of 25 years in Jerusalem. METHODS: Charts of all patients diagnosed as GCA between 1980-2004 were reviewed. In 170 cases GCA was biopsy-proven. Thirty-six additional cases were included as they met the American College of Rheumatology GCA classification criteria. Data on the Jerusalem population throughout the study period was collected from the annual publications of the Israel Bureau of Statistics. Age- and sex-specific incidence rates per 100000 population aged>or=50 were calculated. RESULTS: For the whole period, the average age-adjusted incidence rate was 11.3 per 100000, and 9.5 for the biopsy-positive cases. The female: male ratio was 1.4:1. Cyclic fluctuations of GCA incidence with 3 distinctive peaks, 8 10 years apart, were observed. Altogether, there was no apparent increase in GCA incidence during this period. Seasonal variations were observed: in 192 patients we were able to estimate the time of onset of GCA symptoms. It showed a peak in the months of May and June, with the number of patients being twice as expected for this period (p<0.001). CONCLUSION: GCA onset was more common in late spring and early summer, and fluctuations in GCA annual incidence with 3 distinctive peaks were observed during a 25-year period. These suggest infectious or other environmental etiology, however thus far no such agents were proven. PMID- 17428358 TI - HLA alleles in Korean patients with Takayasu arteritis. AB - In this study, we investigated the HLA allele and haplotype frequencies, and the association of HLA alleles with serious complications and angiographic findings in Korean patients with Takayasu arteritis (TA) compared with healthy controls. Sixty-one patients (56 women, 5 men), diagnosed with TA between January 1995 and December 2005, were studied. Ninety-five healthy men and women were selected as controls. Clinical manifestations were assessed and angiographies were performed at the time of diagnosis in all TA patients. Genotypes of the HLA-A, -B and -DRB1 loci were determined using the polymerase chain reaction-sequencing-based typing (PCR-SBT) method. The mean age at the time of diagnosis of TA was 37.0+/-12.1 years. Compared with controls, the frequencies of A*3001 (p=0.048), B*5201 (p=0.025), and DRB1*1502 (p=0.046) alleles were significantly higher in TA patients, and the frequency of A*2602 was significantly lower in TA patients when compared with controls (p=0.047). The haplotype containing A*2402-B*5201 DRB1*1502 was significantly increased in TA patients (chi2=5.45, p=0.01). Further, among the serious complication of TA, congestive heart failure (CHF) was found to be associated with B*5201 (OR=5.94, p<0.05, 95% CI=1.04 33.85). These data suggest that A*3001, B*5201, and DRB1*1502 alleles might increase the susceptibility to TA, while A*2602 might protect against TA. Further, our results reveal that the haplotype A*2402-B*5201-DRB1*1502 could be a risk factor for TA, and the allele B*5201 is significantly associated with CHF. PMID- 17428359 TI - Adjunction of rituximab to steroids and immunosuppressants for refractory/relapsing Wegener's granulomatosis: a study on 8 patients. AB - OBJECTIVE: Rituximab, an anti-CD20 biotherapy, has been effective against refractory and/or relapsing Wegener's granulomatosis (WG). But the frequency of and time to responses to rituximab, and its effects on various clinical WG manifestations remain to be thoroughly evaluated. METHODS: Retrospective study of 8 patients with refractory/relapsing WG. In addition to their ongoing therapy, 7 patients received rituximab (375 mg/m2 weekly for 4 weeks) and another received 2 rituximab infusions (1 g on days 1 and 15). Disease activity was assessed using BVAS 2003 before and 6 months after the first rituximab infusion. RESULTS: The median BVAS before rituximab was 14.3 (range 4-30). At 6 months, 5/8 patients had BVAS=0; 3/8 were in complete remission; 3/8 in partial remission (lung nodules persisted in 2 patients, scored 0 in BVAS); 2/8 did not respond. One patient relapsed 1 year after stopping rituximab and responded successfully to a second cycle. Dissociated responses of constitutional and 'vasculitis' symptoms, as opposed to granulomatous manifestations, were observed: the former regressed within days or weeks, while the latter regressed more slowly, over several months. Tolerance was good for 7 patients but 1 developed an urticarial rash during the last 3 infusions. Corticosteroids could be tapered in all patients. CONCLUSION: Rituximab, when prescribed in conjunction with corticosteroids and immunosuppressants to treat refractory/relapsing WG, was able to improve clinical outcome. But the dissociation of response times in patients with predominantly granulomatous manifestations, as opposed to vasculitis symptoms, merits further study before an optimal rituximab regimen can be defined. PMID- 17428360 TI - Intravenous immunoglobulins in polyarteritis nodosa restricted to the limbs: case reports and review of the literature. AB - Polyarteritis nodosa (PAN) of the calf muscles is a rare form of vasculitis. We present two cases of PAN limited to the calf and a review of the literature, based on a MEDLINE (PubMed) search of the English literature from 1980 to 2005, using the key words "vasculitis restricted to limbs", "polyarteritis nodosa", and "intravenous immunoglobulin". PAN limited to the calf muscles is a condition presenting with severe shin pain and walking difficulties. In contrast to classic PAN, there is no skin, joint, visceral or nerve system involvement in this form of the disease. The main clinical signs are tenderness and swelling of the calf. Inflammatory markers, such as erythrocyte sedimentation rate and C-reactive protein, are usually elevated, and a perinuclear pattern of anti-neutrophil cytoplasm antibodies can be found. Electromyography of the calf is not contributory. Magnetic resonance imaging may be useful in recognizing the limb restricted vasculopathy and selecting the muscle biopsy site, which is obligatory for diagnosis. Corticosteroids (CS) are the main treatment regimen, but CS resistant cases have been reported. The patients presented here failed to respond to CS but were successfully treated with intravenous immunoglobulin therapy (IVIG). In the absence of vital organ involvement, the addition of cytotoxic drugs is controversial. IVIG seems to be an efficient alternative therapy in PAN limited to the calf muscles especially for patients with limitations to conventional cytotoxic treatment. PMID- 17428361 TI - Low back pain as presenting manifestation of giant cell arteritis associated to abdominal aortitis. AB - Giant cell arteritis (GCA) is the most common vasculitis in Western countries in individuals over the age of 50. The diagnosis is relatively straightforward when typical features, such as headache, jaw claudication or other ischemic complications are present. Although atypical presentations of GCA have been described, herein we report for first time low back pain as the presenting manifestation of this vasculitis. We also emphasize the importance of considering the use of positron emission tomography (PET) in the evaluation of GCA patients presenting without "overt" cranial ischemic manifestations. PMID- 17428362 TI - Gallium-67 scintigraphy in polymyalgia rheumatica. AB - A patient with atypical presentation of polymyalgia rheumatica is presented. Her major symptoms were mild weakness, pain on passive movement of the shoulder and hip girdles, with mild tenderness but no joint stiffness. Muscle enzymes were within the normal range. The diagnostic work-up included gallium-67 scintigraphy, which showed intense uptake in both shoulders, with remarkable improvement following steroid treatment. A prospective study may delineate the possible role of this imaging modality in establishing a diagnosis of polymyalgia rheumatica. PMID- 17428363 TI - Giant-cell arteritis of the female genital tract associated with occult temporal arteritis and FDG-PET evidence of large-vessel vasculitis. AB - We describe a case of giant cell arteritis (GCA) of the female genital tract. Fluorine-18-fluorodeoxyglucose positron emission tomography (18F-FDG PET) and CT scan showed evidence of large-vessel vasculitis involving the thoracic aorta and its branches, while temporal artery biopsy showed arteritis despite the absence of clinical manifestations suggestive of GCA. We review the literature and discuss the relationship between "cranial" GCA, large-vessel GCA and female genital GCA. PMID- 17428364 TI - Lack of association of epithelial cell-derived neurophil-activating peptide (ENA) 78 gene polymorphism with susceptibility to biopsy-proven giant cell arteritis. PMID- 17428365 TI - Churg-Strauss syndrome and Wells syndrome: coincidence or pathogenetic association? A new case report. PMID- 17428366 TI - Endothelium and regulatory cell interactions. AB - Endothelial cells are injury targets in vasculitis and other diseases. However, their abilities to regulate their own fate are becoming increasingly recognized and may influence their susceptibility to injury in different vascular beds. PMID- 17428367 TI - Current knowledge on cellular interactions in the WG-granuloma. AB - Wegener's granulomatosis (WG) usually starts as granulomatous disease of the respiratory tract (so-called localized WG) before it converts to systemic disease (generalized WG) with the emergence of proteinase 3-specific antineutrophil cytoplasmic autoantibodies (PR3-ANCA) and PR3-ANCA associated autoimmune vasculitis. So far, it remains unresolved how tolerance to "Wegener's autoantigen" PR3 is broken and the immune response to PR3 sustained. Further, the relationship between granulomatous lesions and systemic vasculitis is poorly understood. None of the ANCA-animal-models has reproduced granulomata typical of WG so far. A number of endogenous and exogenous factors (HLA DPB1*0401/PTPN22*620W, respiratory epithelial barrier dysfunction? S. aureus, cPR3?) could favour initial formation of granulomata in the respiratory tract and break of tolerance. PR3 induces dendritic cell maturation via the protease activated receptor (PAR)-2 and evokes a strong Th1-type T-cell res-ponse in WG. Clusters of PR3+ cells (neutrophils/monocytes) surrounded by antigen-presenting cells, Th1-type CD4+CD28- effector memory T-cells, maturing B- and plasmacells are found in WG-granulomata of the upper respiratory tract. Thus, WG-granulomata might provide the necessary "proinflammatory environment" for the break of tolerance and display features of lymphoid-like tissue neoformation, in which autoimmunity to PR3 could be sustained. Subsequent PR3-ANCA associated systemic vasculitis gives rise to new inflammatory lesions in many other organs, thereby promoting a self-perpetuating pathology characterized by inflammation and autoimmunity to PR3. PMID- 17428368 TI - The role of pathology in the diagnosis of systemic vasculitis. AB - Pathologic processes are underlying defining features of systemic vasculitis. When these pathologic processes can not be observed directly, surrogate signs and symptoms of disease must be used to conclude that vasculitis is present in a patient and, if so, to determine what specific type of vasculitis is present. This review briefly describes the most defining pathologic features of giant cell arteritis, Takayasu arteritis, polyarteritis nodosa, Henoch-Schonlein purpura, cryoglobulinemic vasculitis, Kawasaki disease, microscopic polyangiitis, Wegener's granulomatosis and Churg-Strauss syndrome; and discusses how these pathologic features can be integrated with clinical and laboratory data to reach an actionable diagnosis. PMID- 17428369 TI - Towards diagnostic criteria for the ANCA-associated vasculitides. PMID- 17428370 TI - Large vessel vasculitis. PMID- 17428371 TI - Imaging studies in the diagnosis of large vessel vasculitis. PMID- 17428372 TI - Giant cell arteritis: recent advances and guidelines for management. AB - This article reviews some of the recent work in epidemiology and pathology of giant cell arteritis (GCA), with particular regard to the immuno-histochemical findings in temporal artery biopsy (TAB) specimens. The diagnostic as well as prognostic role of biopsy histology is discussed. The role of novel imaging techniques e.g. duplex ultrasonography and FDG-PET scanning in diagnosis and staging of disease extent is reviewed. Existing evidence on the treatment is also discussed to propose guidelines on management of GCA. PMID- 17428373 TI - Henoch-Schonlein purpura: recent advances. AB - Recent developments in relation to Henoch-Schonlein purpura (HSP) include: a) a proposed new classification of childhood vasculitides including new classification criteria for HSP; b) the identification of various, potentially important, genetic polymorphisms in HSP that may be relevant in terms of predisposition to or protection from complications; c) evidence that prophylactic steroid at the onset of disease does not protect against renal or gastrointestinal complications but does seem to have beneficial effects in treating them. PMID- 17428374 TI - Lessons learned from an animal model of Kawasaki disease. AB - Kawasaki disease is the most common cause of multisystem vasculitis in childhood. Kawasaki disease has been reported throughout the world and affects children of all ethnicity. Coronary artery damage from Kawasaki disease is the leading cause of acquired heart disease in children in the developed world. Diagnostic tests and prognostic markers are lacking, and questions remain unanswered in our understanding of the etiopathogenesis of the disease, thus limiting our ability to improve therapy and coronary outcome. In this article I will review advances made in an animal model of disease, which has helped advance our understanding of the etiology and pathogenesis of this fascinating clinical syndrome. PMID- 17428375 TI - Part 1: The need for novel treatment regimens for ANCA-associated vasculitis. PMID- 17428376 TI - Part 2: Synopsis of B-lymphocyte targeted therapy of ANCA-associated vasculitis. PMID- 17428377 TI - Part 3: Newer therapies for ANCA-associated vasculitis. PMID- 17428378 TI - [Plasmid encoding human IL-12 improve protective efficacy of Mtb8.4 gene vaccine with signal sequence against infection of Mycobacterium Tuberculosis]. AB - AIM: To study the specific celluar immune response and protective efficacy induced by co-immunization of DNA vaccine of Mtb8.4 with signal sequence (MS) and plasmid encoding human interleukin 12 (hIL-12). METHODS: Fourty C57BL/6N mice were randomly divided into the following groups: MS gene vaccine plus plasmid of hIL-12, MS gene vaccine, BCG, vacant vector alone and PBS. Mice were immunized intramuscularly in both hind limbs three times at the intervals of three weeks or once subcutaneously with 1 x 10(6) of viable M.bovis BCG Pasteur at the time of the first DNA immunization (BCG group). Four weeks after the final inoculation, three mice per group were sacrificed to assess cytokine response by ELISA methods and CTL activities with LDH release assay. The other five mice per group were challenged intravenously in a lateral tail vein with 1 x 10(6) CFU of virulent M.Tuberculosis H37Rv. Spleen and the left lung were harvested from each mouse at 4 weeks after infection and homogenized in sterile. Serial dilutions of organ homogenates were plated on L-J agar and incubated 37 degrees Celsius until colonies were visible 4 weeks later. Protective efficacies in each experiment were expressed as reduced CFU and were compared with that in the negative control group. The right lung was obtained from each mouse and inflated with and stored in 10% formalin immediately. Tissues were embedded in paraffin, sectioned and stained with hematoxylin and eosin. RESULTS: Co-immunization with MS gene vaccine and plasmid encoding hIL-12 induced the secretion of more of Th1 cytokines, but not IL-4, and enhanced CTL activity; while BCG induced the secretion of both types of cytokines (IFN-gamma, IL-2 and IL-4). Co-immunization with MS gene vaccine and plasmid encoding hIL-12 could remarkably reduced CFU counts in organs. CONCLUSION: Plasmid encoding hIL-12 can improve the specific cellular immune response and protective efficacy induced by the MS gene vaccine alone. PMID- 17428379 TI - [Effects of (60)Co gamma rays on the biological behaviour of lung fibroblasts in C57BL/6J and C3H/HeN mice]. AB - AIM: To investigate the effects of (60)Co gamma rays on the biological behaviour of lung fibroblasts (LFs) in C57BL/6J and C3H/HeN mice. METHODS: LFs in C57BL/6J and C3H/HeN mice were isolated, cultured and irradiated by 2, 4, 6 and 8 Gy (60)Co gamma rays. The proliferative activity and cell cycle were detected by MTT colorimetry, AgNOR staining and fluorescence-activated cell sorting(FACS). Then the expressions of alpha-smooth muscle actin (a-SMA), matrix metal proteinase (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1) in LFs were observed by immunocytochemical staining. RESULTS: The proliferation of LFs in C57BL/6J mice was not promoted by 2, 4, 6 and 8 Gy (60)Co gamma rays, but the number of aneuploid cells increased. MMP-1 was weak positive. The expression of a-SMA and TIMP-1 was enhanced by (60)Co gamma rays. The proliferation of LFs in C3H/HeN mice was not promoted by 2, 4, 6 and 8 Gy (60)Co gamma rays either, but the number of cells in G2-M phase increased. The expression of MMP-1 and TIMP-1 was enhanced, but the expression of a-SMA was decreased by (60)Co gamma rays. CONCLUSION: The biological behaviours of the irradiated LFs in C57BL/6J and C3H/HeN mice are different. LFs in C57BL/6J mice can be pactivatedq. Our study will provide a celluar basis for the researches into the C57BL/6J mice which are prone to radiation pulmonary fibrosis (RPF). PMID- 17428380 TI - [Immunity and EGFPC3 expression of recombinant attenuated salmonella typhimurium carrying plasmid pcDNA3s and EGFPC3 in mice]. AB - AIM: To study the immune characterization of HBsAg DNA recombinant attenuated salmonella typhimurium vaccine and the expression of enhanced green fluorescence protein gene pEGFPC3 in mice. METHODS: Plasmid pcDNA3s and pEGFPC3 was transformed into attenuated salmonella typhimurium SL8786 by electricity instrument to construct recombinant vaccine SL8786/EGFPC3 and SL8786/pcDNA3s. Mice were immunized with the recombinant vaccine and the quantity of anti-HBs antibody in the sera was measured by time resolved fluorescence immunoassay(TRFIA), While the cytotoxocity of CTL was measured by LDH release assay. RESULTS: It produced strong specificity CTL response after taking SL8786/pcDNA3s orally to mice and anti-HBs was highest in 11 weeks. EGFPC3 expression in two mice spleen cells was detected by flow cytometry(FCM). EGFP masculine ratio is 19.20% and 17.36% respectively. but SL8786/pcDNA3 in two mice spleens cell EGFP masculine ratio only is 1.95% and 1.63%. The mice was taken SL8786/EGFPC3 orally after three weeks, fluorescence expression in its liver, spleen, kidney, chest gland, duodenum, muscle, etc. were observed. CONCLUSION: HBV DNA vaccine can induce specific cellular and humoral immune responses in vivo in mice by oral. Recombinant attenuated salmonella typhimurium SL8786/EGFPC3 we constructed. It has green fluorescence expression in mice's partial organization, and it offers a model for research gene engineering vaccine. PMID- 17428381 TI - [Isolation, culture and identification of two types of endothelial progenitor cells from peripheral blood in rabbits]. AB - AIM: To investigate how to isolate, culture and identify two types of endothelial progenitor cells from peripheral blood in rabbits. METHODS: Mononuclear cells(MNCs) were isolated from rabbit peripheral blood. Endothelial progenitor cells (EPCs) and endothelial outgrowth cells (EOCs) were obtained from MNCs through different ways of isolation and culture. Two types of cells were assessed by DiI-ac-LDL uptake and lectin binding, and then they were identified by immunofluorescence of flk-1, immunocytochemistry of CD34 and VIII factor related antigen and vasculogenesis activity in vitro. RESULTS: Two types of endothelial progenitor cells were obtained from rabbit peripheral blood through different ways of isolation and culture. EPCs on the seventh day and EOCs on the sixteenth day were positive for ac-LDL uptake and lectin binding, and both of them expressed CD34, flk-1 and VIII factor related antigen. EOCs were assembled into primitive vascular tube-like structures when plated in matrigel. CONCLUSION: EPCs and EOCs could be obtained from rabbit peripheral blood when different ways of isolation and culture were performed. The system of cell culture can be applied to subsequent experiments in cell transplantation. PMID- 17428382 TI - [Construction and expression of fusion gene eukaryotic expression plasmid of pEGFP-C(3)-insig2 and its influence to downstream genes]. AB - AIM: To construct the eukaryotic expression plasmid of insig2 gene and detect the expression of downstream gene adiponectin and adipocyte fatty acid-binding protein 2 (AP2) after the transfection of 3T3-L1 cells. METHODS: Insig2 gene of the mouse was amplified by RT-PCR and then cloned into the eukaryon expression vector pEGFP-C(3), After confirmed by double restriction enzyme digestion analysis and DNA sequencing, pEGFP-C(3)-insig2 was transfected into 3T3-L1 cells by lipofectamine 2000. The expression of insig2 and downstream gene in the 3T3-L1 cells were detected by RT-PCR and fluorescence microscope. RESULTS: The eukaryotic expression plasmid of pEGFP-C(3)-insig2 was constructed. The expression of fusion protein in the endochylema was confirmed. The transcription of adiponectin mRNA and AP2 mRNA was down-regulated after transfection for 24 h and 72 h. CONCLUSION: The eukaryotic expression plasmid of pEGFP-C(3)-insig2 is successfully constructed. The transfected insig2 may have an effect on fat metabolism of 3T3-L1 cells. PMID- 17428383 TI - [Expression of fatty acid binding protein in human breast cancer tissues]. AB - AIM: To detect the expression pattern of fatty acid binding protein (FABP) in human breast cancer, and to find potential markers and therapeutic targets for breast cancer. METHODS: adipocyte-FABP (A-FABP), heart or muscle FABP (H-FABP), brain-FABP (B-FABP), epidermis or psoriasis FABP (E-FABP), liver FABP (L-FABP), intestinal FABP (I-FABP) and gastro-FABP (G-FABP) expression in 35 ductal infiltrating carcinoma and 16 fibroadenoma of breast were detected by RT-PCR, immunohistochemical staining and Western blot analysis, respectively. RESULTS: E , L-, and H-FABP were up regulated significantly in ductal infiltrating carcinoma when compared with those in benign tissue (P<0.05). However, there were not significant difference in A-, B-, G-, and I-FABP expression between ductal infiltrating carcinoma and benign tissue (P>0.05). Interestingly, H-FABP was not only found in benign tissues but in some of ductal infiltrating carcinomas, furthermore, H-FABP level was elevated in malignant tissue, compared with that in benign tissue. CONCLUSION: E-, L-, and H-FABP may play a key role in the progress of invasiveness and metastasis in human breast cancer. Furthermore, the secretion of these FABPs has the potential to serve as a diagnostic marker for breast cancer. PMID- 17428384 TI - [The expression of transforming growth factor beta1 and its I, II receptors in the development of rat embryo and embryonic lung]. AB - AIM: To investigate the expression of TGFbeta1, TbetaRI, TbetaRII in the development of rat embryo and embryonic lung and discuss the interrelationship and function mechanisms between them. METHODS: Half-quantity RT-PCR and immunohistochemical staining were performed in studying these changes. RESULTS: Half-quantity RT-PCR demonstrated that the expressions of these three factors increased in the 13th-15th days and decreased in the 16th-17th days of the embryonic development. Immunohistochemical staining showed that TGFbeta1 was mainly expressed in the developing bronchus epithelial cells.Its receptors were obviously expressed in primitive pulmonary alveolus. CONCLUSION: TGFbeta1 and its receptors play an important regulatory role in the development of rat embryo and embryonic lung, especially in organic morphodifferentiation. PMID- 17428385 TI - [Relationship between TNFalpha and tumor rejection induced by a single dose of melphalan in C57BL/6 mice]. AB - AIM: To investigate the relationship between TNFalpha and tumor rejection induced by a single dose of melphalan in C57BL/6 mice. METHODS: Different gene type mice (TNFR1(+/+), TNFR1(+/-) and TNFR1(-/-)) with the same genetic background of C57BL/6 were used in this experiment. Murine lymphoma EL4 cells were inoculated subcutaneously into the different gene type mice simultaneously. Twelve days later, 7.5 mg/kg melphalan was used intraperitoneally to treat the tumor-bearing mice with TNFR1(+/+), TNFR1(+/-) and TNFR1(-/-). The tumors in the different gene type mice were observed and recorded every one to three day. RESULTS: After the treatment of 7.5 mg/kg melphalan during the first week, the tumors in the different gene type mice shrank at a similar rate. In the following 2 months, the tumors in the TNFR1(+/+) and TNFR1(+/-) C57BL/6 mice gradually shrank and were cured but most tumors in the TNFR1(-/-) C57BL/6 mice relapsed after melphalan treatment. CONCLUSION: TNFalpha plays an important role in melphalan-induced tumor rejection. The anti-tumor effect of melphalan has no relationship with the expression of tumor necrosis factor 1 in tumor-bearing mice. TNFR1 is required to prevent or avoid the relapse of tumors in mice instead of tumor cells. PMID- 17428386 TI - [Expression of Langerhans cell-related chemokine genes in condyloma acuminatum epidermis]. AB - AIM: To investigate differential gene expression of Langerhans cell-related chemokines in condyloma acuminatum (CA) epidermis and normal epidermis. METHODS: Gene expression of Langerhans cell-related chemokines in three CA epidermal lesions and in three normal controls was screened using Affymetrix oligonucleotide microarrays HG-U 133A 2.0, and part of the above differential gene expression was confirmed by semi-quantitative RT-PCR. RESULTS: With microarrays, seven down-regulated genes of Langerhans cell-related chemokine were detected in three CA epidermal lesions as compared with three normal controls, and the down-regulation of CXCR4 and CCL20 was verified by semi-quantitative RT PCR. CONCLUSION: Several Langerhans cell-related chemokine genes are found down regulated in CA epidermis as compared with normal epidermis, and the down regulation of these genes may contribute to the decreased number and the homing disturbance of LC in CA epidermis. PMID- 17428387 TI - [Effect of Rapamycin on CD4(+) CD25(+) regulatory T cells in allo-transplantation tolerance model]. AB - AIM: To study the effect of Rapamycin(Rapa) on CD4(+) CD25(+) regulatory T cells in allo-transplantation tolerance model. METHODS: The model of skin allo transplantation was established. The recipient BALB/c mice were injected with allogeneic donor spleen cells from B6 on the day before grafting and with cyclosporine(CsA) after transplantation. T cells were purified on the day 14 from the tolerant group and cultured with Rapa and/or IL-2 in different concentrations in vitro. Mixed lymphocyte reaction (MLR) was used to analyze the specific recall reaction of T cells. Percentages of CD4(+) CD25(+) regulatory T cells were examined by flow cytometry (FCM). The expression of Foxp3 mRNA was measured by RT PCR, and the level of IL-10 in supernatant of T cells cultured in vitro was determined by ELISA. BALB/c-SCID mice underwent allo-transplantation were given adoptive transfer of T cells treated with Rapa and/or IL-2, then the survival conditions of the grafted skin were observed daily. RESULTS: CsA plus donor splenocyte injection can prolong allograft of BALB/c significantly (P<0.05). The number of CD4(+) CD25(+) regulatory T cells and the expression of Foxp3 mRNA for T cell in the tolerant group treated with Rapa and/or IL-2 were obviously increased. Adoptive transfusion of T cells from tolerant mice treated with IL 2/Rapa obviously prolonged the allograft survival time in allografted-SCID mice. CONCLUSION: Rapa can increase the ratio of CD4(+) CD25(+) regulatory T cells in vitro and prolong the graft survival time obviously after adoptive immunity, and these effects are enhanced by low-dose of IL-2. PMID- 17428388 TI - [The changes of CD4(+) CD25(high) regulatory T cells in the course of rheumatoid arthritis and their significances]. AB - AIM: To explore the percentages of CD4(+) CD25(high) regulatory T cells in peripheral blood or synovial fluid from patients with rheumatoid arthritis (RA) on different stages, and to study their correlations with the activity of the disease. METHODS: Peripheral blood lymphocytes were obtained from the patients with active RA who had received no previous disease modifying (DMARDs) therapy(n=11), from individuals with stable, well-controlled RA (n=12), from subjects whose disease were poorly improved after DMARDs therapy(n=9), and from healthy controls(n=8). The frequencies of CD4(+) CD25(high) T cells were quantified by using flow cytometry(FCM). Meanwhile, the correlations between the percentage of CD4(+) CD25(high) T cells and the level of Anti-CCP antibody, CRP, ESR and RF were also investigated. Paired blood and synovial fluid was analysed in a small group of RA and other patients. RESULTS: There was a smaller proportion of CD4(+) CD25(high) T cells in the peripheral blood of the active RA patients(mean 5.24%) and poorly-improved RA patients(mean 6.43%) than in patients with stable well-controlled RA or in healthy controls(mean 11.79% and 17.17%, respectively, P<0.01 in each case). The frequency of CD4(+) CD25(high) T cells from RA was negatively associated with Anti-CCP antibody(58.0 Ru/mL), ESR(38.8 mm/h) and CRP(2.73 mug/L), (P<0.05 for each). On contrast, The frequency of CD4(+) CD25(high) T cells from healthy controls was not significantly correlated with the level of Anti-CCP antibody(<5.0 Ru/mL), ESR(4.67 mm/h), CRP(0.15 mug/L) and RF(1.37) (all P>0.1). The percentage of CD4(+) CD25(high) regulatory T cells from synovial fluid of RA patients (24.32%) was significantly lower than that of AS patients(30.24%) (P<0.05). CONCLUSION: The results demonstrate a smaller CD4(+) CD25(high) regulatory T cell population in peripheral blood of individuals with active RA prior to disease modifying treatment and with poorly-improved RA, and this is negatively associated with the activity of the disease. PMID- 17428389 TI - [Preparation and characterization of human colon tumor-associated antigen and its clinical significance]. AB - AIM: To purify the colon tumor-associated antigen from cultured colon tumor cells, and to investigate its expression in the sera of patients with colon cancer. METHODS: The monoclonal antibody (mAb) against human colon tumor associated antigen 4D10 was employed as the ligand for the immunoaffinity chromatography to purify the colon tumor-associated antigen from the lysate of colorectal tumor cell LOVO. The purified antigen was identified by SDS-PAGE and Western blot. The expression of colon tumor-associated antigen in the sera of patients with colon cancer and in normal sera was detected by Sandwich ELISA. RESULTS: The purified colon tumor-associated antigen binding to mAb 4D10 was a heterodimer composed of two subunits with relative molecular mass M(r) of 30 x 10(3) and 35 x 10(3) respectively. The antigen was significantly higher expressed in sera from patients with colon cancer than that in normal sera (P<0.01). CONCLUSION: The tumor-associated antigen obtained from the colon tumor cells has been successfully purified through immunoaffinity chromatography with mAb 4D10, which may be useful for diagnosis on clinic. PMID- 17428390 TI - [Enhancement of HSP-MUC1 antitumor activity by type C CpG-ODN BW005]. AB - AIM: To investigate the enhancement of anti-tumor effect of HSP-MUC1 by self designed type C CpG-ODN BW005. METHODS: The immunostimulatory effect of CpG-ODN BW005 was detected by IFN protection assay and (3)H proliferation assay in vitro. Sixty C57BL/6 mice were separated into 5 groups randomly, including Sodium Chloride control, HSP-MUC1 control, HSP-MUC1/1585, HSP-MUC1/1826 and HSP MUC1/BW005. Mice were injected s.c. with agents on day 0, 14 and 28 and were implanted MUC1-EL4 tumor cells s.c. on day 33. Tumor growth and murine death were recorded. Blood was collected in 57 day from tail vein. Subtype of anti-HSP and anti-MUC1 IgG in serum was detected by indirect ELISA. RESULTS: CpG-ODN BW005 could stimulate the proliferation of hPBMC and mice spleoncyte and IFNalpha production. HSP-MUC1/BW005 postponed tumor development, with the average tumor developed day of 44.8, and prolonged the survival of mice with the average survival day of 49.5. Moreover, final tumor-developed rate of this group was 33.33%, which was the lowest; final survival rate of this group was 66.67%, which was the highest. Levels of anti-HSP and anti-MUC1 IgG2a in HSP-MUC1/CpG-ODNs group were enhanced. CONCLUSION: CpG-ODN BW005, a kind of type C CpG-ODN, could enhance the anti-tumor effect of HSP-MUC1. PMID- 17428391 TI - [Myocardial autoimmune response induced by myosin activate T lymphocytes]. AB - AIM: To investigate whether myosin activated-T lymphocytes induce myocardial autoimmune response. METHODS: Dendritic cells and T lymphocytes were isolated and purified from spleens of Lewis rats. After cocultured in the absence or presence of cardiac myosin (20 mug/mL), the sensitized or unsensitized T lymphocytes were transferred into the naive syngeneic Lewis rats by a single tail vein infusion, respectively. The recipient rats were killed on the third day, at the first and fourth week and then the histopathological changes of the rats were observed. RESULTS: T lymphocyte infiltration was observed on the third day. It reached the peak with occasional myocardium necrosis at the first weekend. T cell infiltration distinctly lessened at the fourth weekend. No T lymphocyte infiltration was found in kidney, liver, lung, and brain. T lymphocyte infiltration wad not detected in the heart of the transferred unsensitized T lymphocytes of the rats. CONCLUSION: Myocardial autoimmune response can be induced by myosin activated T lymphocytes. PMID- 17428392 TI - [Lipopolysaccaride influenced peritoneal mesothelial cells on the growth and expression of tumor necrosis factor through nuclear factor kappaB pathway]. AB - AIM: To investigate whether lipopolysaccharide (LPS) influence peritoneal mesothelial cells (PMCs) on their apoptosis, growth and expression of tumor necrosis factor (TNF) through IkappaBalpha, nuclear factor p65 pathway. METHODS: PMCs were isolated, cultured and passaged by enzymatic disaggregation, then identified by phase contrast inverted microscope, transmission electron microscope and scanning electron microscope, with immunocytochemistry method. The PMCs were treated under conditions, which included different concentration LPS (normal control group, 0.1 mg/L, 1.0 mg/L, 10 mg/L, 50 mg/L, 100 mg/L) and different time (0, 1, 3, 6, 12, 24 hours). Hoechst 33258 fluorescence dye method was used to detect the apoptosis of PMC. The inhibitory effect of LPS on cell growth was measured with MTT method. Immunofluorescence dye was used to observe the activity of p65. Western blot method was used to detect the expression of IkappaBalpha. L929 cell line was used to detect the activity of TNF. RT-PCR was used to measure the expression of TNFalpha mRNA. RESULTS: LPS could inhibit growth of PMC and induce apoptosis. LPS could lead IkappaBalpha to degration and p65 into nucleus. LPS could also induce the expression of TNF. The expression of TNF-alpha mRNA reached the peak at the 1st hour and highest TNF activity at the 3rd hour. CONCLUSION: LPS inhibit the growth of PMC and induced the apoptosis. LPS induced PMC to express TNF-alpha in a dose-dependent manner. In the early state, PMC expressed a great deal of TNF-alpha, then began to fall quickly. During the period that LPS influence PMC on apoptosis, growth and expression of TNF, IkappaBalpha-NF-kappaB p65 pathway played the important role. PMID- 17428393 TI - [Prepare and study on the characterization of a new monoclonal antibody to germ tube Candida albicans]. AB - AIM: To prepare and identify the monoclonal antibody(mAb) specific for the germ tube of Candida albicans. METHODS: mAb were prepared via hybridoma technique. Indirect immunofluorescence (IIF) was used to analyze the activity and specificity of the mAb. The protection of the mAb was analyzed via the inhibition of the mAb to the germ tube formation and the adhesion of C.albicans to the epithelial and endothelial cells. The frozen hybridoma cells were cultured to analyze the ability of the secretion of the mAb. RESULTS: mAb03.2C1-C2 belongs to IgG1 subclass. Relative molecular mass (M(r)) of antigens recognized by the mAb was 156, 48, 22 and 12. The target antigen could be detected 30 minutes after the germ tube formation. The epitope might lie in the N-carbohydrate chain. mAb03.2C1 C2 was shown by IIF test to be specific to the surface of the germ tube or mycelium phase of C.albicans. In the protective mAb screening experiment, it was found that mAb3.2C1-C2 could inhibit C.albicans germ tube formation and the adhension of C.albicans to the epithelial and endothelial cells. CONCLUSION: In vitro, mAb03.2C1-C2 could inhibit the formation of germ tube of C.albicans, which could depress the invasiveness of C.albicans. This anti-C.albicans cell wall surface-specific mAb may be a good candidate for use in tests for the rapid differentiation of the two closely related species C.albicans and C.dubliniensis. PMID- 17428394 TI - [Preparation of the monoclonal antibodies against human BORIS and the expression pattern of BORIS in normal and diseased human mammary tissues]. AB - AIM: To prepare monoclonal antibodies(mAb) to BORIS antigen and analyze its expression pattern in breast cancer, benign breast disease and normal breast tissues. METHODS: Female BALB/c mice were immunized with recombination human BORIS protein. Hybridoma cell lines were established by hybridoma technique. BORIS antigen was detected by immunohistochemical (ICH) staining. RESULTS: Four clones of hybridomas stably secereting mAb against human BORIS were obtained. mAb FMMU-BORIS13 was selected to be employed in ICH. Positive staining was localized in the nuclei of the spermatocytes and spermatogoniums, which was consistent with the typical expression pattern of CT antigen. BORIS antigen was detected in 85%(94/110) of breast cancer, 95%(21/22) of benign breast diseases and 6 normal breast tissues(surrouding tumor free breast tissue). CONCLUSION: Four clones of anti-BORIS mAb have been successfully prepared. The expression of BORIS antigen is much stronger in breast cancer than that in benign breast disease and normal breast tissues, which indicate that BORIS may be involved in the pathogenesis of the breast cancer. PMID- 17428395 TI - [Preparation and characterization of monoclonal antibodies against human CD112 (Nectin2/PRR2)]. AB - AIM: To express and purify fusion protein of human CD112 extracellular region, prepare monoclonal antibodies (mAb) to CD112 and investigate the distribution of CD112 molecule. METHODS: The CD112-Fc fusion protein was expressed with gene recombinant technique in eukaryotic system and purified with affinity chromatography column. The BALB/c mice were immunized with purified CD112-Fc for preparation of mAb by hybridoma technique. The prepared mAbs were identified by indirect ELISA, Western blot and flow cytometry (FCM). Subsequently, the distribution of CD112 on different human cell lines was investigated by FCM. RESULTS: The effective expression plasmid pIg3C-CD112 was constructed, and human CD112-Fc was successfully expressed and purified with a purity of more than 90%. Nine clones of hybridoma were obtained, which were designated as FMU-CD112.1-FMU CD112.9. FMU-CD112.1, 3, 6 and 8 specifically reacted with recombinant CD112-Fc protein in Western blot and FMU-CD112.1, 4, 6 and 8 could be used in FCM. The investigation of CD112 distribution showed high expression on the cell lines differentiated from epithelial cells. CONCLUSION: The hybridomas secreting mAbs to human CD112 are established successfully, which may lay the foundation for further research on the biological function of CD112. PMID- 17428396 TI - [Prokaryotic expression and purification of RPS13 and preparation of polyclonal antibody against RPS13]. AB - AIM: To construct prokaryotic expression plasmid of RPS13, express and purify the protein for the preparation of polyclonal antibody. METHODS: RPS13 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from the highly expressed cell of gastric multidrug cell SGC7901/VCR. After sequenced, the gene was cloned into the expression vector pET-28a(+) to construct RPS13 expression plasmid pET-28a(+)-RPS13. The expression plasmid was transformed into the E.coli BL21 and screened by ampiline, and then the E.coli BL21 containing the expression plasmid was induced by IPTG and the protein was purifed by Histag column. BALB/c mice were immunized by the immunogen His-RPS13. The titer was measured by ELISA and the polyclonal antibody was obtained. RESULTS: The expression plasmid pET-28a(+)-RPS13 was constructed. The 19 kDa recombined protein was successfully expressed in the E.coli BL21 by IPTG for 3 hours, purified by Histag column and confirmed by SDS-PAGE and Western blot. The polyclonal anti-His-RPS13 antibody was obtained by immunizing the mice with RPS13 protein. RPS13 could be specifically recognized by the antibody based on Western blot. CONCLUSION: RPS13 has bene expressed and purified successfully and polyclonal anti-His-RPS13 antibody has been prepared. Our study can be used for the preparation of monoclonal anti-His-RPS13 antibody and for further research of the function of the RPS13 protein in tumor. PMID- 17428397 TI - [Effect of low serum culture on the synchrony of cell cycle of mesenchymal stem cells]. AB - AIM: To investigate a suitable method of inducing cell cycle synchronization at G0/G1 stage of mesenchymal stem cells (MSCs). METHODS: MSCs were cultured and identified with CD44, CD90, CD71 and CD11b by flow cytometer. Cell cycle and apotosis under normal and low serum culture were detected by flow cytometer. RESULTS: MSCs were positive for CD44, CD90 and CD71 and negative for CD11b. The cells at G0/G1 cell stage decreased, while the cells at S and G2 cell stage increased in 1 day of 50 mL/L fetal bovine serum (FBS) and 1 to 3 days of 5 mL/L FBS. However, prolonged culture in serum-starvation medium induced cell cycle arrest at G0/G1 stage. The ratio of apoptotic cells did not increase in 50 mL/L FBS. In 5 mL/L FBS, the ratio of apoptotic cells increased in 3 days and then decreased in 4 to 5 days. The proportion of cells at G0/G1 phase was significantly increased from 75.9% to 89.4% while the ratio of apoptotic cells was only 0.162% in 5 days of 5 mL/L FBS. CONCLUSION: It is a good method to induce the synchrony of cell cycle of MSCs at G0/G1 stage in five days of 5 mL/L FBS. PMID- 17428398 TI - Climatic change and asthma. PMID- 17428399 TI - Specific immunotherapy is effective in the prevention of new sensitivities. AB - BACKGROUND: Allergen specific immunotherapy is the only specific and curative approach in the treatment of IgE-mediated allergic diseases such as bronchial asthma and allergic rhinitis. The safety and clinical efficacy of this treatment are well documented but data on the prevention of new sensitizations remain scarce. OBJECTIVE: To demonstrate the efficacy of specific immunotherapy in allergic respiratory diseases in childhood and to determine whether this treatment prevents the development of new sensitizations in children sensitized to house dust mite or pollen species. METHODS: Fifty-six patients received specific immunotherapy (43 sensitive to house dust mite and 13 sensitive to pollen). Fifty-one patients not receiving immunotherapy but treated with pharmacotherapy were enrolled in the control group. The patients were followed-up for at least 4 years and treatment efficacy and the development of new sensitizations were compared between the two groups. RESULTS: The number of patients with symptoms was significantly decreased in both groups at the end of the treatment period. No new sensitizations were found in 35 of the 43 (81.39 %) patients in the house dust mite immunotherapy group and in 10 of 13 (76.92 %) patients in the pollen immunotherapy group. In contrast, 20 of 51 (39.21 %) patients in the control group showed new sensitizations. The difference between the house dust mite and pollen immunotherapy groups and the control group in this parameter was statistically significant (p = 0.033). CONCLUSION: Our data demonstrate that administration of specific immunotherapy in allergic patients significantly reduced symptoms and the development of new sensitivities. PMID- 17428400 TI - Therapeutic effects of the anti-tumor necrosis factor monoclonal antibody, infliximab, in four children with refractory juvenile idiopathic arthritis. AB - OBJECTIVE: To report the results of treatment with infliximab in patients with refractory juvenile idiopathic arthritis (JIA). PATIENTS AND METHODS: A prospective study of four children with refractory JIA was carried out. Infliximab (100 mg) was administered in weeks 0, 2 and 6. Subsequently, the drug was administered every 8 weeks. The following parameters were assessed at the beginning and at the end of the follow-up period: number of joints with active arthritis, number of joints with a limited range of motion, physician overall assessment of disease activity, parent assessment of the child's overall well being, pain assessment scores, and erythrocyte sedimentation rate. Improvement was rated according to the definition of the American College of Rheumatology (ACR 30). Paired sample tests were used for statistical analysis. RESULTS: Three girls and one boy aged between 10 and 16 years old with a history of JIA ranging from 1 to 9 years were included. The patients received infliximab for a period of 11 to 33 months (average 22 months). There was a significant decrease in the number of swollen joints (p < 0.05), joints with a limited range of movement (p < 0.04), pain score assessment (p < 0.005), physician overall assessment (p = 0.002), maternal evaluation (p < 0.001), the patient's own evaluation (p < 0.001), and duration of morning stiffness (p < 0.001). Both steroids and methotrexate dosages were reduced and no adverse effects or infections were registered. CONCLUSIONS: Infliximab improved joint inflammatory indexes and clinical assessments. This improvement increased the quality of life of the patients and their families, suggesting that the use of biological therapy is a good option in refractory JIA. PMID- 17428401 TI - Chronic urticaria: what is new, where are we headed. AB - Chronic urticaria can be defined as the occurrence of widespread daily or nearly daily wheals for at least 6 weeks, which may be accompanied by angioedema. It is a disease with a considerable impact on patients' quality of life. Furthermore, these patients may undergo extensive laboratory evaluations seeking a cause only to be frustrated when none is found. There is no curative treatment for this disorder and we do not understand the mechanisms that lead to the onset of disease. However, in recent years there have been significant advances in the understanding of some of the molecular mechanisms that cause cutaneous inflammation that is manifest as urticaria and angioedema. In this review we will summarize our recent contributions to this field and try to offer insights regarding future directions for research. PMID- 17428402 TI - Intensity of bronchial hyperresponsiveness and asthma relapse risk in the young adult. AB - BACKGROUND: The evolution of asthma starting in childhood varies and depends on a series of factors (atopy, allergens, and environmental irritants, etc). Treatment may influence the evolution of the disease and even cause the symptoms to disappear. However, there remains a risk of relapse years later. OBJECTIVES: To assess the role of bronchial hyperresponsiveness in asthma relapse in young adulthood in patients with symptoms that disappeared after treatment prescribed in childhood. MATERIAL AND METHODS: To determine the evolution of asthma and patients' personal opinions, 78 patients were sent a questionnaire several years after having been discharged without symptoms in the previous 2 years, and without the need for medication. The methacholine test was used to evaluate bronchial hyperresponsiveness at discharge. The 40 patients who correctly completed the questionnaire were divided into three groups according to the methacholine dose required to obtain a 20 % decrease in forced expiratory volume in 1 second (PD20): group 1 (15 patients), < 1000microg; group 2 (10 patients) between 1001 and 2000 microg; and group 3 (15 patients) > 2100 microg. The mean age at discharge was 16 years (range 13-25 years) versus 26 years at the time of response (range 18-33 years), with a similar distribution in all three groups. Age at disease onset, with estimation of severity, age at the first visit and at the start of treatment, and respiratory function were evaluated. RESULTS: Thirty of the interviewed patients considered themselves to be cured. Seven of the patients (three in group 1, one in group 2, and three in group 3) did not consider themselves to be cured, although their symptoms were minimal and they rarely used medication. Health status was described as "regular" with sporadic symptoms by one patient in each group. No correlation with methacholine response was observed. CONCLUSION: No relationship was found between the degree of bronchial hyperresponsiveness and the risk of relapse in young adults who suffered asthma in childhood. PMID- 17428403 TI - Partial response to cyclosporine in a patient with Schnitzler's syndrome. AB - Schnitzler's syndrome is an unusual clinical association of chronic urticaria, intermittent fever and monoclonal immunoglobulin M (IgM) gammopathy. The pathogenesis of the urticaria is unclear and treatment is problematic. We describe the case of a 61-year-old woman with a long history of chronic urticaria with severe pruritus, spiking fever and malaise. The IgM-kappa monoclonal component was detected in the patient's serum 4 years after symptom onset. After ineffective treatment with antihistamines and systemic corticosteroids, oral cyclosporine resulted in complete remission of the fever and malaise, which has persisted after an 18-month follow-up. Partial but maintained remission of the urticaria was also observed, allowing corticosteroid doses to be decreased. PMID- 17428404 TI - Cross-reactivity between Parietaria pollen and beet. AB - Allergy to beet is very rare. Until now, only a few reports about asthma induced by inhaling the vapor of cooked beet have been published. We describe two patients with allergic rhinitis and positive skin prick tests to Parietaria and beet only. To investigate possible cross-reactivity between Parietaria pollen and beet, we performed laboratory assays that showed beet-specific IgE in the sera of both patients and possible cross-reactivity between Parietaria and beet in one patient. PMID- 17428405 TI - Re: a prospective safety study of allergen immunotherapy in daily clinical practice. PMID- 17428406 TI - [Difficulties understanding compressed speech in elderly listeners with mild-to moderate hearing loss]. AB - OBJECTIVE: In this work we evaluated the difficulties in understanding rapid speech (normal, fast and very fast rates) in elderly listeners with and without hearing loss (presbycusis and moderate hearing loss). Rapid speech is common in daily communication, yet few studies have been conducted to assess this problem in Spanish-speaking listeners, as has been done for English speakers. MATERIAL AND METHOD: We compared the recognition of sentences presented at normal, fast, and very fast speech rates in 3 groups of listeners. Recognition scores were correlated to their audiological measures and the score obtained on a Communication Difficulties Questionnaire. RESULTS: Compressed sentences were more poorly perceived in the 2 groups of listeners with hearing loss, compared to the normal group. Moreover, we obtained significant correlations among the 3 measures: recognition of rapid speech, the audiological measures, and the score in the Questionnaire. CONCLUSIONS: Our results suggest that rapid-speech recognition skills deteriorate in the presence of hearing loss, whether due to presbycusis or other moderate hearing loss in all frequency ranges. PMID- 17428407 TI - [Treatment of epistaxes in hereditary haemorrhagic telangiectasia (Rendu-Osler Weber disease) with tranexamic acid]. AB - OBJECTIVE: Recurrent epistaxis is the most frequent clinical manifestation of hereditary haemorrhagic telangiectasia (HHT). Its treatment occasionally presents difficulties as there is no consensus on the appropriate therapeutic protocol. Our objective was to explore the utility of oral tranexamic acid for the treatment of epistaxes in HHT patients. PATIENTS AND METHOD: A 3-year prospective study was carried on HHT patients with epistaxis treated with oral tranexamic acid in the HHT unit at our hospital. RESULTS: Ten patients with HHT were treated with oral tranexamic acid during the study. Most of them improved both the frequency and severity of their epistaxis and were satisfied with the treatment. No treatment-related complications were recorded. Two patients needed more aggressive treatments to control epistaxis. CONCLUSIONS: Oral tranexamic acid is useful for achieving significant reductions in epistaxis frequency and intensity in selected patients with HHT. In those presenting severe epistaxis, however, it may need to be combined with more aggressive therapies. PMID- 17428408 TI - [Facial nerve repair techniques]. AB - OBJECTIVE: Post-operative facial function was analyzed in 14 patients undergoing different techniques for facial nerve repair. MATERIAL AND METHOD: The aetiology of facial paralysis included vestibular schwannomas, facial schwannomas, cranioencephalic trauma, and iatrogenic lesions. RESULTS: The mean interval between paralysis and reconstruction surgery was 6 months, although in 2 cases it exceeded 12 months. The first facial signs of recovery were achieved around the sixth month post-operatively, in every patient. All patients achieved a facial function graded III or IV in the House-Brackmann scale, equivalent to a grade II or III on the May scale. There were no differences between the surgical techniques compared, both termino-terminal anastomosis and hypoglossal-facial anastomosis. CONCLUSIONS: The surgical reconstruction techniques for facial nerve repair should be known by all ENT surgeons. At least facial symmetry at rest and complete eye closure are achieved in almost every patient when the procedure is carried out at the proper time. PMID- 17428409 TI - [Autoimmune hearing loss: improving diagnostic performance]. AB - OBJECTIVES: Due to the lack of specific serological markers for the diagnosis of immune-mediated hearing loss, an exhaustive immunologic work-up study for patients is usually performed. The aim of the present study is to find the most cost-effective laboratory tests used for the diagnosis of this entity. PATIENTS AND METHOD: Comparative study between 2 groups of patients with a high suspicion of suffering diverse clinical forms of immune-mediated hearing loss, subjected to different serologic testing designs: the classical immunologic work-up study (125 patients) in comparison with a more restricted examination analysis (57 patients), based on a high risk profile recently reported. RESULTS: Diagnostic efficiency was similar. CONCLUSIONS: Since financial resources are limited, ANA and immunophenotype of PBL (peripheral blood lymphocytes) are recommended for the evaluation of a patient with suspected immune-mediated hearing loss. PMID- 17428410 TI - [Conventional audiometry versus cochlear microphone audiometry]. AB - OBJECTIVE: The recording and processing of cochlear microphone potentials in hearing studies is currently in the definitive validation phase against results obtained with other objective procedures. The purpose of this work is to contribute to its validation. MATERIAL AND METHOD: The equipment used was exclusively designed for recording cochlear microphone potentials. The study has been carried out in adults to compare subjective audiometric results with those obtained from cochlear microphones. RESULTS: We present a statistical concordance study between subjective audiometry and cochlear microphone audiometry. CONCLUSIONS: In view of the results obtained, this method is particularly valid for early diagnosis. We obtained an identical profile to the subjective audiogram at audiometric frequencies of 250, 500, 1000, 2000, and 4000 Hz. PMID- 17428411 TI - [Effect of irrigation sequence in vestibular paresis results of bithermal caloric tests]. AB - OBJECTIVE: Bithermal caloric tests are among those most commonly used in vestibular function studies and require a properly systematized approach. Irrigation sequence varies between laboratories. Our objective is to study the effect of irrigation sequence on the percentage of vestibular paresis. MATERIAL AND METHOD: We analysed 1214 records, 77.51% cold to hot sequences (C-H) and the rest hot to cold (H-C). We considered results above 24% of vestibular asymmetry to be pathological. RESULTS: In general, 33.36% of the records were pathological. Groups compared showed that H-C sequence had more significant (P <0.001) pathological results than C-H (46.52% vs 29.54%). CONCLUSION: Irrigation sequence has a significant influence on the outcome of caloric tests to for the detection of vestibular paresis. More studies are needed to confirm the best irrigation sequence. PMID- 17428412 TI - [Tumour recurrence in squamous head and neck cancer]. AB - INTRODUCTION: For most patients with squamous head and neck cancer (HN-SCC), locoregional tumour recurrence (TR) carries an extremely poor prognosis and is therapeutically challenging. OBJECTIVE: To define the clinical aspects of TR and their implication on the survival in patients with HN-SCC. PATIENTS AND METHOD: The clinical management and the outcome of 652 patients with HN-SCC were reviewed. RESULTS: The overall incidence of TR in this series of HN-SCC was 19.9% (n=130). The most frequent locations of the primary cancers were oropharynx (32%), hypopharynx (24%), and oral cavity (21%). The rates of recurrence were locoregional 50%, local 43 % and stomal recurrence 7%. The appearance of a TR reduces the overall survival of patients with HN-SCC to 15%. Survival is better in glottic (38%) and supraglottic (27%), carcinomas, and worse in oro-hypopharynx tumours (2-4%). CONCLUSIONS: RT are more frequent in pharyngeal tumours, especially locoregional recurrences. Patients with recurrence in pharynx were definitely associated with poor prognosis and in these cases salvage surgery seems not to improve survival rates. PMID- 17428413 TI - [Invasive sinusal mycosis due to Chrysosporium tropicum]. AB - An invasive sinusal mycosis in a patient with acquired immunodeficiency is reported. The clinical course and causal agent (Chrysosporium tropicum) were both rather atypical. This fungus has been seldom isolated as a cause of nasosinusal infections. Just one more case like this is found when literature is reviewed. We emphasize importance of early diagnosis and treatment to reduce morbi-mortality. PMID- 17428414 TI - [Calcification of auricular cartilages in adrenal insufficiency]. AB - Calcification of the auricular cartilages is a very rare finding observed in metabolic disorders like Addison's disease, adrenal insufficiency or diabetes. Its aetiology and physiopathological mechanisms remain unclear. It may be caused by exposure to cold or trauma in the cartilage but in a significant number of cases, ossification occurs without the identification of the precipitating cause or event. We present the clinical and pathologic findings of a patient suffering from Addison's disease who developed auricular bilateral calcification. The literature is reviewed and the physiopathological hypotheses discussed. PMID- 17428415 TI - [Schwannoma of the external auditory canal: an exceptional location]. AB - Solitary schwannoma is a benign, encapsulated tumour of Schwann cell origin, therefore the olfactory and optic nerves are never affected. About 25%-45% of all schwannomas occur in the head and neck but schwannoma of the external auditory canal is a rare finding, and we have found only 6 previous cases reported in the literature world-wide. Our patient was discovered by chance during a stapedectomy because the tumour was sited in the external auditory canal without distorting it. PMID- 17428416 TI - [Specific qualifications in phoniatry: consequences of their abandonment]. PMID- 17428417 TI - [First Spanish Conference on Health Prevention and Promotion in Clinical Practice: an inter-professional and inter-disciplinary collaborative effort]. PMID- 17428418 TI - [No more ''clicking'' on boxes]. PMID- 17428419 TI - [Initial treatment of patients with subjective memory complaints and/or cognitive impairment in primary care: ISSEA Study]. AB - OBJECTIVE: To describe the initial treatment of patients >60 years who had subjective memory complaints and/or cognitive impairment for at least 6 months. DESIGN: Observational, longitudinal, multicentre, and naturalistic study, with a follow-up period of 12 months. SETTING: A total of 105 primary care centres. PARTICIPANTS: The study included 921 patients who attended a clinic. MAIN MEASUREMENTS: In the baseline visit, the social demographic characteristics, diagnosis, treatment, and the health care referrals were recorded, and in the next 2 (6 and 12 months) the diagnostic change, treatment, and referrals. RESULTS: The majority of subjects were female (66.9%) and the mean age was 74.3+/ 6.8 years. In the initial visit, 50.5% (95% confidence interval [CI], 47.3-53.7) of the diagnoses were classed as syndromic and 33.3% (95% CI, 30.3-36.3), aetiological. The primary care doctor modified an unconfirmed initial diagnosis in 22% (95% CI, 19.3-24.7) of the patients during the 12 months follow-up study. A diagnosis was made in 63.8% (95% CI, 60.7-66.9) of patients by anamnesis, physical examination, the screening test, and laboratory data. In the initial visit, el 52.6% (95% CI, 49.4-55.8) received treatment of mainly nootropics and neuroprotectors, and later, the percentage of patients on cholinesterase inhibitors increased. The primary care doctor referred 54.9% (95% CI, 51.7-58.1) of the patients during the 12 months of the study. CONCLUSIONS: The primary care doctor diagnoses the majority of patients with the means available, mainly based on anamnesis, examination, the screening test, and laboratory data. They prescribe drugs that lack efficacy for this disease and few of those that have been shown to be effective. PMID- 17428420 TI - [Commentary: cognitive impairment: where are we?]. PMID- 17428421 TI - [Usefulness of a glaucoma and ocular hypertension screening strategy in primary care]. AB - OBJECTIVES: To evaluate the usefulness of a glaucoma and intraocular hypertension screening strategy for new cases detected. To evaluate the acceptability of taking intra-ocular pressure (IOP) and the appearance of side effects. DESIGN: Cross-sectional, descriptive study. SETTING: An urban health centre and the ophthalmology clinic of its main hospital. PARTICIPANTS: A total of 2044 patients aged over 40, 63.5% women and 36.5% men, with a mean age of 61.23 (SD, 11.42). They were selected by consecutive sampling from patients who visited the health centre over a 9-month period. Subjects diagnosed with glaucoma, ocular hypertension (OH), conjunctivitis, or corneal pathology were excluded. INTERVENTIONS: Taking of IOP with Tonopen XL in primary care. Subjects with IOP > or = 21 mm Hg were referred to ophthalmology. In these patients, IOP was measured with Goldmann, and patients with confirmed OH received ophthalmoscopy and campimetry. MAIN MEASUREMENTS: Percentage of subjects with glaucoma, suspected glaucoma, and OH confirmed in ophthalmology. Positive predictive value (PPV) for OH. RESULTS: One hundred subjects with OH were detected (4.89%; 95% CI, 3.93% 5.85%), of whom 21 were diagnosed with glaucoma (1.04%; 95% CI, 0.57-1.49) and 10 with suspected glaucoma (0.49%; 95% CI, 0.16-0.82). The PPV for OH was 44.27%. The acceptability of the test was 98.09%. No patients presented with side-effects following the taking of their IOP. CONCLUSIONS: The strategy evaluated is useful in terms of the number of subjects with glaucoma and OH detected. The acceptability of taking IOP with Tonopen XL was high. PMID- 17428422 TI - [Short Zarit interview on burden of care for caregivers in primary care]. AB - OBJECTIVE: To determine which Short Zarit Interview behaves best for use in establishing caregivers' burden in primary care. DESIGN: Study of validation of diagnostic tests. LOCATION: Vilaboa Health Centre, Pontevedra, Spain. PARTICIPANTS: Main caregivers of elderly dependent patients who live at home. INTERVENTIONS: Katz Index to determine dependence, Zarit Burden of Care Interview and palliative care version of the Short Zarit Interview, Bedard, Bedard screening version, and the Japanese version. MAIN MEASUREMENTS: Zarit Interview in its 22-item version, with subsequent calculation of the sensitivity (S), specificity (Sp), positive predictive value (PPV), and negative predictive value (NPV) of the various reduced versions. RESULTS: The Short Zarit Interview used in palliative care cases to determine the family giving up has an S of 100%, Sp 90.5%, PPV 95.45%, and NPV 100% in defining caregivers' burden in primary care. CONCLUSIONS: The scale that behaves best in primary care is the Short Zarit Interview for palliative care. PMID- 17428423 TI - [Detection of adolescents at risk of suffering eating disorders]. AB - OBJECTIVES: To analyse which risk factors are related to eating disorders and to identify useful questions for their early detection in primary care. DESIGN: Observational, cross-sectional study. SETTING: Six secondary schools in Leganes, Madrid, Spain. PARTICIPANTS: Adolescents aged between 12 and 18 years old attending secondary school. MAIN MEASUREMENTS: Risk of suffering an eating disorder (Eating Attitudes Test-26) and questions about weight, height, family, attitudes towards body image, use of health services for eating disorders, and mental illness history. RESULTS: An 8.8% of the adolescents were at risk of developing an eating disorder (4.6% male; 11.8% female). Age in males (OR=0.76) and 24-hour fasting to lose weight (OR for males =7.44; OR for females =3.09) were associated with the risk of eating disorders. Early menarche (OR=1.69), having dieted for social or environmental reasons, and self-induced vomiting (OR=5.71) were associated with eating disorders in women. The specificity and negative predictive value of asking about self-induced vomiting and 24-hour fasting in order to detect an eating disorder were around 95%. CONCLUSIONS: Age is a protective factor for eating disorders, especially in males. In women, early menarche is a risk factor. Asking about self-induced vomiting and 24-hour fasting enables eating disorders in women to be ruled out. PMID- 17428424 TI - [Management systems for the renewal of medical prescriptions: the vision of users and health professionals]. AB - OBJECTIVE: To find the degree of satisfaction of users and doctors who use the repeat medical prescription system. DESIGN: Multi-centre, cross-sectional study. SETTING: Four primary care centres covering 160 000 inhabitants. PARTICIPANTS: Users included in the repeat medical prescription system and doctors who use it. MAIN MEASUREMENTS: Questionnaire on satisfaction. MAIN RESULTS: Users (52.4% women) filled in 429 questionnaires (mean age, 64.2; SD, 13.98). Average medicines taken were 4.4 (SD, 3.04). The period established for picking up prescriptions was usually 60 days (77.4%). 83.9% (95% CI, 80.4-87.4) of those questioned stated they had no problems, with a mean degree of satisfaction at 8.4 (SD, 1.52). There were no significant differences for gender (P=.53). There was a very weak but significant positive correlation between the score and the age of those questioned (r(2)=1%; P=.03). Mean score of satisfaction had statistically significant differences, depending on the PCC. 86.9% (95% CI, 83.8-90.1) of users collected the prescriptions within the term set. The factors associated with greater adherence were age and a greater number of medicines per person. Doctors filled in 47 questionnaires. Mean score was 7.42 (SD, 1.09) and all except one (98%; 95% CI, 89.1-99.9) thought that the system meant an improvement in clinical management. CONCLUSION: Users are very satisfied, although they believe certain organisational points should be improved in order to improve accessibility. PMID- 17428425 TI - [Cardiovascular PAPPS 2005: a critical analysis]. PMID- 17428426 TI - [Family medicine in Colombia]. PMID- 17428427 TI - [Clinical and therapeutic management of respiratory tract infections. Consensus document of the Andalusian infectious diseases society and the Andalusian family and comunitary medicine society]. AB - Respiratory tract infections are frequent and they are one of the commonest causes of antibiotic prescription. However, there are few clinical guidelines that consider this group of infections. This document has been written by the Andalusian Infectious Diseases Society and the Andalusian Family and Community Medicine Society. The primary objective has been to define the recommendations for the diagnosis and antibiotic treatment of respiratory tract infections apart from pneumonia. The clinical syndromes evaluated have been: a) pharyngitis; b) sinusitis; c) acute otitis media and otitis externa; d) acute bronchitis, laryngitis, epiglottitis; e) acute exacerbation of chronic bronchitis; and f) respiratory infectious in patients with bronchiectasis. This document has focused on immunocompetent patients. PMID- 17428428 TI - [BODE Index, can we classify COPD better in primary care?]. PMID- 17428429 TI - [Comorbidity recorded in patients diagnosed with fibromyalgia at a primary care centre]. PMID- 17428430 TI - [Lights and shadows of rural medicine: concerning teaching]. PMID- 17428431 TI - [Professionals' access to drug package inserts]. PMID- 17428432 TI - Genotyping of Giardia duodenalis from human and animal samples from Brazil using beta-giardin gene: a phylogenetic analysis. AB - Giardia duodenalis is one of the major diarrhea agents in human and animals distributed worldwide, and present high levels of genetic diversity, showing seven genotypes: A, B, C, D, E, F, and G. Only Assemblages A and B have been detected in humans and in a wide range of other mammalians hosts, whereas the remaining Assemblages (C-G) are host-specific. Molecular characterization of cysts of human and animal origin are useful to address the co-circulate isolates between these host, and represents an objective means to evaluate zoonotic infection hypothesis. In the present work the G. duodenalis genotypes were characterized by restriction fragment length polymorphisms and DNA sequencing analysis of PCR products of the beta-giardin gene. The cysts were collected in the city of Rio de Janeiro, in Brazil, from a population composed by humans (n=366, 310 children and 56 adults), domestic animals (n=11) from a municipal daycare center in the surroundings of a slum and neighborhood medium-high class domestic animals (n=18). Parasitological exams were developed in human fecal samples. Parasites were found in 60% (186/310) and 66% (37/56) of the samples from children and adults, respectively. Among children's samples, 27.7% (86/310) were positive for G. duodenalis. Only 1.7% (1/56) of the adults was positive for this parasite. In general a total of 87 fecal samples (86 from children and 1 from adult) from all population studied were positive for G. duodenalis, and 62 of these were subjected to molecular analysis using a PCR that amplified a fragment of the beta-giardin gene. Sixty samples were typed as genotype A1, two as genotype A2 and genotype B was not encountered. Among domestic animals samples (n=29), eight (seven dogs and one cat) from the slum community were identified as genotype A1, and all control samples (n=18) were negative in the molecular assay. The host-specific genotypes C, D and, F were not found. In this study we described single case of G. duodenalis infection associated with a child and her dog and both isolates characterized as genotype A1. Despite the low incidence, this data suggest the putative existence of a zoonotic cycle of G. duodenalis in the studied population. PMID- 17428433 TI - Nanocomposite of functionalized multiwall carbon nanotubes with nafion, nano platinum, and nano gold biosensing film for simultaneous determination of ascorbic acid, epinephrine, and uric acid. AB - A unique bimetallic, nano platinum (Pt) with nano gold (Au) on nafion (NF) incorporated with functionalized multiwall carbon nanotubes (f-MWCNTs) composite film (f-MWCNTs-NF-PtAu) was developed by the potentiostatic method. The composite film exhibits promising efficient catalytic activity towards the oxidation of mixture of biochemical compounds and simultaneous measurement of ascorbate anion, epinephrine and urate anion in aqueous buffer solution (pH 6.75). Both, the cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used for the measurement of electroanalytical properties of neurotransmitters by means of composite film modified electrodes. Well-separated voltammetric peaks were obtained for ascorbate, epinephrine and urate anions with the peak separations of 0.222 and 0.131V. The composite film can also be produced on gold and transparent semiconductor indium tin oxide electrodes for different kinds of studies such as electrochemical quartz crystal microbalance (EQCM), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The incorporation of Pt and Au onto the f-MWCNTs-NF was revealed by the EQCM technique and the morphology of the film was studied using SEM, AFM and scanning electrochemical microscopy (SECM) techniques. Further, extensive studies were carried out using SECM for obtaining the surface current topographic images of composite film modified electrodes, and these indicated the presence of f-MWCNTs-NF-PtAu composite film on the electrode. PMID- 17428434 TI - Crystal structure of an aerobic FMN-dependent azoreductase (AzoA) from Enterococcus faecalis. AB - The initial critical step of reduction of the azo bond during the metabolism of azo dyes is catalyzed by a group of NAD(P)H dependant enzymes called azoreductases. Although several azoreductases have been identified from microorganisms and partially characterized, very little is known about the structural basis for substrate specificity and the nature of catalysis. Enterococcus faecalis azoreductase A (AzoA) is a highly active azoreductase with a broad spectrum of substrate specificity and is capable of degrading a wide variety of azo dyes. Here, we report the crystal structure of the AzoA from E. faecalis determined at 2.07 A resolution with bound FMN ligand. Phases were obtained by single wavelength anomalous scattering of selenomethionine labeled protein crystals. The asymmetric unit consisted of two dimers with one FMN molecule bound to each monomer. The AzoA monomer takes a typical NAD(P)-binding Rossmann fold with a highly conserved FMN binding pocket. A salt bridge between Arg18 and Asp184 restricts the size of the flavin binding pocket such that only FMN can bind. A putative NADH binding site could be identified and a plausible mechanism for substrate reduction is proposed. Expression studies revealed azoA gene to be expressed constitutively in E. faecalis. PMID- 17428435 TI - Kinetic variations determine the product pattern of phytoene desaturase from Rubrivivax gelatinosus. AB - In bacteria and fungi, the degree of carotenoid desaturation is determined by a single enzyme, the CrtI-type phytoene desaturase. In different organisms, this enzyme can carry out either three, four or even five desaturation steps. The purple bacterium Rubrivivax gelatinosus is the only known species in which reaction products of a 3-step and a 4-step desaturation (i.e. neurosporene and lycopene derivatives) accumulate simultaneously. The properties of this phytoene desaturation to catalyze neurosporene or lycopene were analyzed by heterologous complementations in Escherichia coli and by in vitro studies. They demonstrated that high enzyme concentrations or low phytoene supply favor the formation of lycopene. Under these conditions, CrtI from Rhodobacter spheroides can be forced in vitro to lycopene formation although this carotene is not synthesized in this species. All results can be explained by a model based on the competition between phytoene and neurosporene for the substrate binding site of phytoene desaturase. Mutations in CrtI from Rvi. gelatinosus have been generated resulting in increased lycopene formation in Escherichia coli. This modification in catalysis is due to increased amounts of CrtI protein. PMID- 17428436 TI - Characterisation and biological activity of Glu3 amino acid substituted GIP receptor antagonists. AB - Glucose-dependent insulinotropic polypeptide (GIP) is an important gastrointestinal hormone, which regulates insulin release and glucose homeostasis, but is rapidly inactivated by enzymatic N-terminal truncation. Here we report the enzyme resistance and biological activity of several Glu(3) substituted analogues of GIP namely; (Ala(3))GIP, (Lys(3))GIP, (Phe(3))GIP, (Trp(3))GIP and (Tyr(3))GIP. Only (Lys(3))GIP demonstrated moderately enhanced resistance to DPP-IV (p<0.05 to p<0.01) compared to native GIP. All analogues demonstrated a decreased potency in cAMP production (EC(50) 1.47 to 11.02 nM; p<0.01 to p<0.001) with (Lys(3))GIP and (Phe(3))GIP significantly inhibiting GIP stimulated cAMP production (p<0.05). In BRIN-BD11 cells, (Lys(3))GIP, (Phe(3))GIP, (Trp(3))GIP and (Tyr(3))GIP did not stimulate insulin secretion with both (Lys(3))GIP and (Phe(3))GIP significantly inhibiting GIP-stimulated insulin secretion (p<0.05). Injection of each GIP analogue together with glucose in ob/ob mice significantly increased the glycaemic excursion compared to control (p<0.05 to p<0.001). This was associated with lack of significant insulin responses. (Ala(3))GIP, (Phe(3))GIP and (Tyr(3))GIP, when administered together with GIP, significantly reduced plasma insulin (p<0.05 to p<0.01) and impaired the glucose lowering ability (p<0.05 to p<0.01) of the native peptide. The DPP-IV resistance and GIP antagonism observed were similar but less pronounced than (Pro(3))GIP. These data demonstrate that position 3 amino acid substitution of GIP with (Ala(3)), (Phe(3)), (Tyr(3)) or (Pro(3)) provides a new class of functional GIP receptor antagonists. PMID- 17428437 TI - Multi-site inhibition of human plasma cholinesterase by cationic phenoxazine and phenothiazine dyes. AB - Two cationic phenoxazine dyes, meldola blue (MB) and nile blue (NB), and the structurally related phenothiazine, methylene blue (MethB), were found to act as complex inhibitors of human plasma cholinesterase (butyrylcholinesterase, BChE). Studied at 25 degrees C, in 100mM MOPS buffer (pH 8.0), with butyrylthiocholine as substrate, the kinetic pattern of inhibition indicated cooperative I binding at 2 sites. Intrinsic K' values ( identical with[I](0.5)(2) extrapolated to [S]=0) for MB, NB and MethB were 0.64+/-0.05, 0.085+/-0.026 and 0.42+/-0.04 microM, respectively. Under the same experimental conditions the dyes acted as single-occupancy, hyperbolic-mixed inhibitors of electric eel acetylcholinesterase (AChE), with K(i)=0.035+/-0.010, 0.026+/-0.0034 and 0.017+/ 0.0063 microM (for MB, NB, MethB); alpha (coefficient of competitive interaction)=1.8-2.4 and beta (coefficient of noncompetitive interaction)=0.15 0.28. The complexity of the BChE inhibitory effect of phenoxazine/phenothiazine dyes contrasted with that of conventional ChE inhibitors which cause single occupancy (n=1), competitive or mixed inhibition in both AChE and BChE and signaled novel modes of ligand interaction at (or remote from) the active site gorge of the latter enzyme. PMID- 17428438 TI - Modulation of in vitro and in vivo angiogenesis by alternagin-C, a disintegrin like protein from Bothrops alternatus snake venom and by a peptide derived from its sequence. AB - We have previously demonstrated that alternagin-C (ALT-C), a disintegrin-like protein from the venom of the Brazilian snake Bothrops alternatus, induces human vascular endothelial cell (HUVEC) proliferation by up-regulating the expression of vascular endothelial growth factor (VEGF). Here, we show that ALT-C is also able to induce in vivo angiogenesis using the model of matrigel plug in nude mice. Fibroblast growth factor (FGF) alone or supplemented with ALT-C was mixed with melted matrigel and subcutaneously injected in nude mice. After two weeks, the matrigel plugs were removed and analyzed to verify endothelial cell migration and new vessel formation. ALT-C (1 and 10 ng) strongly induced endothelial cell migration as well as the formation of new vessels. However, in higher concentrations, ALT-C strongly inhibited angiogenesis. In low concentrations (1 and 10nM), ALT-C also up-regulates the expression of VEGF receptor 2 (VEGFR2, KDR) mostly after 48 h, but it did not affect VEGFR1 (Ftl-1) in HUVEC cells as demonstrated by real-time PCR analysis. However, in higher concentrations (100 nM) the expression of both receptors is down-regulated. A peptide derived from ALT-C primary structure also affects HUVEC proliferation in vitro and angiogenesis in vivo. In conclusion, the present study shows for the first time the in vivo angiogenesis induced by a disintegrin-like molecule and the modulation of VEGFRs as well. PMID- 17428439 TI - Reaction pattern and mechanism of light induced oxidative water splitting in photosynthesis. AB - This mini review is an attempt to briefly summarize our current knowledge on light driven oxidative water splitting in photosynthesis. The reaction leading to molecular oxygen and four protons via photosynthesis comprises thermodynamic and kinetic constraints that require a balanced fine tuning of the reaction coordinates. The mode of coupling between electron (ET) and proton transfer (PT) reactions is shown to be of key mechanistic relevance for the redox turnover of Y(Z) and the reactions within the WOC. The WOC is characterized by peculiar energetics of its oxidation steps in the WOC. In all oxygen evolving photosynthetic organisms the redox state S(1) is thermodynamically most stable and therefore this general feature is assumed to be of physiological relevance. Available information on the Gibbs energy differences between the individual redox states S(i+1) and S(i) and on the activation energies of their oxidative transitions are used to construct a general reaction coordinate of oxidative water splitting in photosystem II (PS II). Finally, an attempt is presented to cast our current state of knowledge into a mechanism of oxidative water splitting with special emphasis on the formation of the essential O-O bond and the active role of the protein environment in tuning the local proton activity that depends on time and redox state S(i). The O-O linkage is assumed to take place within a multistate equilibrium at the redox level of S(3), comprising both redox isomerism and proton tautomerism. It is proposed that one state, S(3)(P), attains an electronic configuration and nuclear geometry that corresponds with a hydrogen bonded peroxide which acts as the entatic state for the generation of complexed molecular oxygen through S(3)(P) oxidation by Y(Z)(ox). PMID- 17428440 TI - Both chlorophylls a and d are essential for the photochemistry in photosystem II of the cyanobacteria, Acaryochloris marina. AB - We have measured the flash-induced absorbance difference spectrum attributed to the formation of the secondary radical pair, P(+)Q(-), between 270 nm and 1000 nm at 77 K in photosystem II of the chlorophyll d containing cyanobacterium, Acaryochloris marina. Despite the high level of chlorophyll d present, the flash induced absorption difference spectrum of an approximately 2 ms decay component shows a number of features which are typical of the difference spectrum seen in oxygenic photosynthetic organisms containing no chlorophyll d. The spectral shape in the near-UV indicates that a plastoquinone is the secondary acceptor molecule (Q(A)). The strong C-550 change at 543 nm confirms previous reports that pheophytin a is the primary electron acceptor. The bleach at 435 nm and increase in absorption at 820 nm indicates that the positive charge is stabilized on a chlorophyll a molecule. In addition a strong electrochromic band shift, centred at 723 nm, has been observed. It is assigned to a shift of the Qy band of the neighbouring accessory chlorophyll d, Chl(D1). It seems highly likely that it accepts excitation energy from the chlorophyll d containing antenna. We therefore propose that primary charge separation is initiated from this chlorophyll d molecule and functions as the primary electron donor. Despite its lower excited state energy (0.1 V less), as compared to chlorophyll a, this chlorophyll d molecule is capable of driving the plastoquinone oxidoreductase activity of photosystem II. However, chlorophyll a is used to stabilize the positive charge and ultimately to drive water oxidation. PMID- 17428441 TI - Prediction of subcellular protein localization based on functional domain composition. AB - Assigning subcellular localization (SL) to proteins is one of the major tasks of functional proteomics. Despite the impressive technical advances of the past decades, it is still time-consuming and laborious to experimentally determine SL on a high throughput scale. Thus, computational predictions are the preferred method for large-scale assignment of protein SL, and if appropriate, followed up by experimental studies. In this report, using a machine learning approach, the Nearest Neighbor Algorithm (NNA), we developed a prediction system for protein SL in which we incorporated a protein functional domain profile. The overall accuracy achieved by this system is 93.96%. Furthermore, comparisons with other methods have been conducted to demonstrate the validity and efficiency of our prediction system. We also provide an implementation of our Subcellular Location Prediction System (SLPS), which is available at http://pcal.biosino.org. PMID- 17428442 TI - Magneto immuno-PCR: a novel immunoassay based on biogenic magnetosome nanoparticles. AB - We describe an innovative modification of the Immuno-PCR technology for automatable high sensitive antigen detection. The Magneto Immuno-PCR (M-IPCR) is based on antibody-functionalized biogenic magnetosome nanoparticles revealing major advantages over synthetic magnetic particles. The general principle of the M-IPCR is similar to that of a two-sided (sandwich) immunoassay. However, antibody-functionalized magnetosome conjugates were employed for the immobilization and magnetic enrichment of the signal generating detection complex enabling the establishment of a surface independent immunoassay. To this end, the M-IPCR was carried out by simultaneously tagging the antigen with the reagent for read-out, i.e., a conjugate comprising the specific antibody and DNA fragments, in the presence of the antibody-functionalized magnetosomes. To demonstrate the general functionality of the M-IPCR, the detection of recombinant Hepatitis B surface Antigen (HBsAg) in human serum was established. We observed a detection limit of 320pg/ml of HBsAg using the M-IPCR, which was about 100-fold more sensitive than the analogous Magneto-ELISA, established in parallel for comparison purposes. PMID- 17428443 TI - Incorporation of low molecular weight molecules into alpha(2)-macroglobulin by nucleophilic exchange. AB - alpha(2)-Macroglobulin (alpha(2)M) is a proteinase inhibitor that functions by a trapping mechanism which has been exploited such that the receptor-recognized, activated form (alpha(2)M( *)) can be employed to target antigens to antigen presenting cells. Another potential use of alpha(2)M( *) is as a drug delivery system. In this study we demonstrate that guanosine triphosphate, labeled with Texas red (GTP-TR) formed complexes with alpha(2)M( *) following activation by proteolytic or non-proteolytic reactions. Optimal incorporation occurred with 20 microM GTP-TR, pH 8.0 for 5h at 50 degrees C. NaCl concentration (100 or 200 mM) had little effect on incorporation at this pH or temperature, but was significant at sub-optimum temperature and pH values. Maximum incorporation was 1.2 mol GTP TR/mol alpha(2)M( *). PAGE showed that 70-90% of the GTP-TR is bound in a SDS/2 mercaptoethanol resistant manner. Guanosine, adenosine, and imidazole competed with GTP-TR to form complexes with alpha(2)M( *). PMID- 17428444 TI - Caldesmon inhibits the actin-myosin interaction by changing its spatial orientation and mobility during the ATPase activity cycle. AB - Orientation and mobility of acrylodan fluorescent probe specifically bound to caldesmon Cys580 incorporated into muscle ghost fibers decorated with myosin S1 and containing tropomyosin was studied in the presence or absence of MgADP, MgAMP PNP, MgATPgammaS or MgATP. Modeling of various intermediate states of actomyosin has shown discrete changes in orientation and mobility of the dye dipoles which is the evidence for multistep changes in the structural changes of caldesmon during the ATPase hydrolysis cycle. It is suggested that S1 interaction with actin results in nucleotide-dependent displacement of the C-terminal part of caldesmon molecule and changes in its mobility. Thus inhibition of the actomyosin ATPase activity may be due to changes in caldesmon position on the thin filament and its interaction with actin. Our new findings described in the present paper as well as those published recently elsewhere might conciliate the two existing models of molecular mechanism of inhibition of the actomyosin ATPase by caldesmon. PMID- 17428445 TI - Factors affecting the substrate specificity of histone deacetylases. AB - Histone deacetylases (HDACs) catalyze the deacetylation of epsilon-acetyl-lysine residues within the N-terminal tail of core histones and thereby mediate changes in the chromatin structure and regulate gene expression in eukaryotic cells. So far, surprisingly little is known about the substrate specificities of different HDACs. Here, we prepared a library of fluorogenic tripeptidic substrates of the general format Ac-P(-2)-P(-1)-Lys(Ac)-MCA (P(-1), P(-2)=all amino acids except cysteine) and measured their HDAC-dependent conversion in a standard fluorogenic HDAC assay. Different HDAC subtypes can be ranked according to their substrate selectivity: HDAH > HDAC8 > HDAC1 > HDAC3 > HDAC6. HDAC1, HDAC3, and HDAC6 exhibit a similar specificity profile, whereas both HDAC8 and HDAH have rather distinct profiles. Furthermore, it was shown that second-site modification (e.g., phosphorylation) of substrate sequences as well as corepressor binding can modulate the selectivity of enzymatic substrate conversion. PMID- 17428446 TI - An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone. AB - Mechanistic aberrations leading to Gemcitabine (2',2'-dFdCyd,2,2 difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance. PMID- 17428447 TI - L-454,560, a potent and selective PDE4 inhibitor with in vivo efficacy in animal models of asthma and cognition. AB - Type 4 phosphodiesterases (PDE4) inhibitors are emerging therapeutics in the treatment of a number of chronic disorders including asthma, chronic obstructive pulmonary disease (COPD) and cognitive disorders. This study delineates the preclinical profile of L-454,560, which is a potent, competitive and preferential inhibitor of PDE4A, 4B, and 4D with IC50 values of 1.6, 0.5 and 1.2 nM, respectively. In contrast to the exclusive binding of cilomilast and the preferential binding of roflumilast to the PDE4 holoenzyme state (Mg2+-bound form), L-454,560 binds to both the apo-(Mg2+-free) and holoenzyme states of PDE4. The intrinsic enzyme potency for PDE4 inhibition by L-454,560 also results in an effective blockade of LPS-induced TNFalpha formation in whole blood (IC50 = 161 nM) and is comparable to the human whole blood potency of roflumilast. The cytokine profile of inhibition of L-454,560 is mainly a Th1 profile with significant inhibition of IFNgamma and no detectable inhibition of IL-13 formation up to 1 microM. L-454,560 was also found to be efficacious in two models of airway hyper-reactivity, the ovalbumin (OVA) sensitized and challenged guinea pig and the ascaris sensitized sheep model. Furthermore, L-454560 was also effective in improving performance in the delayed matching to position (DMTP) version of the Morris watermaze, at a dose removed from that associated with potential emesis. Therefore, L-454,560 is a novel PDE4 inhibitor with an overall in vivo efficacy profile at least comparable to roflumilast and clearly superior to cilomilast. PMID- 17428448 TI - Mechanism of Ca(2+)/calmodulin-dependent protein kinase IV activation and of cyclic AMP response element binding protein phosphorylation during hypoxia in the cerebral cortex of newborn piglets. AB - Previously we showed that hypoxia results in increased neuronal nuclear Ca(2+) influx, Ca(2+)/calmodulin-dependent protein kinase IV activity (CaM KIV) and phosphorylation of c-AMP response element binding (CREB) protein. The aim of the present study was to understand the importance of neuronal nuclear Ca(2+) in the role of CaM KIV activation and CREB protein phosphorylation associated with hypoxia. To accomplish this the present study tests the hypothesis that clonidine administration will block increased nuclear Ca(2+) influx by inhibiting high affinity Ca(2+)/ATPase and prevent increased CaM KIV activity and CREB phosphorylation in the neuronal nuclei of the cerebral cortex of hypoxic newborn piglets. To accomplish this piglets were divided in three groups: normoxic, hypoxic, and hypoxic-treated with clonidine. The piglets that were in the Hx+Cl group received clonidine 5 min prior to hypoxia. Cerebral tissue hypoxia was confirmed biochemically by tissue levels of ATP and phosphocreatine (PCr). The data show that clonidine prevents hypoxia-induced increase in CaM KIV activity and CREB protein phosphorylation. We conclude that the mechanism of hypoxia induced activation of CaM KIV and CREB phosphorylation is nuclear Ca(2+) influx mediated. We speculate that nuclear Ca(2+) influx is a key step that triggers CREB mediated transcription of apoptotic proteins and hypoxic mediated neuronal death. PMID- 17428449 TI - Altered cholesterol metabolism in APP695-transfected neuroblastoma cells. AB - Cholesterol has been implicated to play an important role in the generation of Abeta peptides, which are the main component of beta-amyloid plaques in the brains of patients suffering from Alzheimer's disease (AD). Epidemiological data implicate that lowering cholesterol levels has beneficial effects on the extent of beta-amyloid pathology. Thus therapeutic intervention using cholesterol lowering drugs like statins seems to be a promising approach. A couple of studies, in vitro or in vivo by the use of AD transgenic mouse models, focused on the manipulation of cholesterol levels and the resulting effects on Abeta generation. In contrast, there is not much known about the effect of the amyloid precursor protein (APP) on cholesterol levels. In the present report, we transfected human neuroblastoma cells with human APP695 and compared cellular cholesterol levels with the respective levels in Mock-transfected control cells. Furthermore, we determined the levels of diverse cholesterol precursors and metabolites using gas chromatography-mass spectrometry (GC-MS). Significant differences in the levels of the respective cholesterol precursors were observed, whereas inhibition of gamma-secretase activity by the gamma-secretase inhibitor DAPT did not have a significant effect on cellular cholesterol metabolism. PMID- 17428450 TI - Trigemino-cardiac reflex during transsphenoidal surgery for pituitary adenomas: methodological description of a prospective skull base study protocol. AB - A systematic clinical neuroscience protocol is described for the use to examine the trigemino-cardiac reflex (TCR) response in humans. Target neurosurgical conditions are operations that require manipulations around the peripheral and central part of the trigeminal nerve and its branches, e.g. the cerebellopontine angle or the sellar region. To assess the hemodynamic and cardiac responses of patients after TCR initiation, anesthetic monitoring has been applied. The TCR is defined as a drop of more than 20% of the heart rate and the mean arterial blood pressure compared with the baseline values before the stimulus and coinciding with the surgical manipulation at or around any branches of the trigeminal nerve. By help of illustrative cases, we present for the first time preliminary results regarding the differentiation of the TCR in a central and a peripheral induction during transsphenoidal surgery of pituitary adenomas. Based on these results, we can conclude that we have developed a battery of preoperative examination procedures based on event-related diagnostics that was useful to differentiate different subgroups of TCR during transsphenoidal surgery. The presented protocol can be performed directly pre-, intra- and postoperatively and applied for assessment of TCR even in patients with known risk factors. PMID- 17428451 TI - ERP evidence for successful voluntary avoidance of conscious recollection. AB - We investigated neurocognitive processes of voluntarily avoiding conscious recollection by asking participants to either attempt to recollect (the Think condition) or to avoid recollecting (the No-Think condition) a previously exposed paired associate. Event-related potentials (ERPs) during Think and No-Think trials were separated on the basis of previous learning success versus failure. This separation yielded temporal and topographic dissociations between early ERP effects of a Think versus No-Think strategy, which were maximal between 200 and 300 ms after stimulus presentation and independent of learning status, and a later learning-specific ERP effect maximal between 500 and 800 ms after stimulus presentation. In this later time-window, Learned Think items elicited a larger late left parietal positivity than did Not Learned Think, Learned No-Think, and Not Learned No-Think items; moreover, Learned No-Think and Not Learned Think items did not differ in late left parietal positivity. Because the late left parietal positivity indexes conscious recollection, the results provide firm evidence that conscious recollection of recollectable information can be voluntarily avoided on an item-specific basis and help to clarify previous neural evidence from the Think/No-Think procedure, which could not separate item specific from strategic processes. PMID- 17428452 TI - Cellular localization of mGluR3 and mGluR5 mRNAs in normal and injured rat brain. AB - In order to understand the role of metabotropic glutamate receptors (mGluRs) in the brain, it is important to know how the mGluRs are differentially expressed among the different cell types. At present, the cellular expression of mGluR3 and mGluR5 has been mostly studied in terms of proteins with observations suggesting the expression of both mGluR3 and mGluR5 in neuronal and in glial cells. In order to verify the brain cell type-expressing mGluR3 and mGluR5 mRNAs, both in normal and injured brain, we performed a double labeling analysis, by in situ hybridization for mGluR3 or mGluR5 mRNA and immunohistochemistry for specific cellular markers. This approach allowed us to find mGluR3 mRNA expressed in neurons (NeuN-positive cells), and in glial cells, such as astrocytes (GFAP positive cells) and oligodendrocytes (CNPase-positive cells). The same analysis showed that only NeuN-positive cells express mGluR5 mRNA. The time course of mGluR3 mRNA expression in two models of hippocampal formation lesion, kainate induced seizures or ibotenic acid injection, showed an increased expression of mGluR3 in the area of lesion. This effect appears 1 week after the injury and was localized in GFAP- and CNPase-positive cells. In contrast, mGluR5 was not found expressed in the area of lesion. The present results contribute to extend available data on cell type-expressing mGluR3 and mGluR5 in normal and injured brain and could be relevant to understand the mechanisms that drive neuron-glial cells interaction both in normal and repairing processes. PMID- 17428453 TI - Infarct volume after transient middle cerebral artery occlusion (MCAo) can be reduced by attenuation but not by inactivation of c-Jun action. AB - Stroke therapy aims to save penumbral tissue from apoptosis that is activated in response to the ischemic injury. Since the c-Jun transcription factor plays a crucial role in promoting apoptosis, inhibition of its activation might reduce the final infarct size and thus increase functional outcome. To test this hypothesis we made use of four genetically modified mouse lines influencing the c Jun pathway at various steps. Upon transient middle cerebral artery occlusion for 90 min and 24 h of reperfusion, infarct volume and number of ATF-2-, TUNEL- and cleaved Caspase-3-positive cells were determined in conditional c-Jun knock-out mice (cond. c-Jun), mice overexpressing JunB (JunBtg), mice lacking the phosphoacceptor serines 63 and 73 of c-Jun (JunAA) and in mice overexpressing Bcl 2 (Bcl-2tg). Cond. c-Jun as well as JunAA mice did not show significant differences in the infarct size when compared to their non-mutant controls. By contrast smaller infarct volumes were detected in transgenic mice merely attenuating c-Jun action (JunBtg and Bcl-2tg). ATF-2, TUNEL or cleaved Caspase-3 staining revealed no significant differences between the experimental groups. A complete lack of functional c-Jun might be compensated by other cellular mechanisms, in contrast to its reduced function. Thus, our data suggest that attenuation rather than a complete block of c-Jun action appears to be more promising for therapy of stroke. PMID- 17428454 TI - Chorda tympani nerve transection impairs the gustatory detection of free fatty acids in male and female rats. AB - Sprague-Dawley rats with intact (SHAM) and bilaterally transected chorda tympani nerves (CTX) received conditioned taste aversions (CTAs) to the free fatty acids (FFAs), linoleic and oleic acid, at micromolar quantities. Two-bottle preference tests showed that CTX eliminated avoidance of 88 muM linoleic acid but did not affect CTA avoidance of corn oil or 250 mM sucrose. Short-duration stimulus tests following single-pairing CTAs revealed that 8-s stimulus durations resulted in higher detection thresholds for linoleic acid than 30-s trials. In these short duration tests, CTX rats showed 2-fold elevations in threshold for linoleic acid compared to the SHAM rats. A single-pairing CTA did not produce avoidance of oleic acid during the short-duration tests; however, 3 consecutive days of CTA pairings did produce avoidance of oleic acid in both male and female rats. Finally, both male and female rats received SHAM or CTX surgery after demonstrating successful CTAs to either 100 microM linoleic or oleic acid. The ability to detect and avoid linoleic and oleic acid was eliminated by CTX for both sexes. Differences in the ability of rats to form CTAs to linoleic and oleic acid suggest that linoleic acid is a more salient stimulus than oleic acid. Our results suggest that FFAs stimulate afferent taste signals in the chorda tympani nerve of male and female rats and that these signals play an important role in the gustatory behavior of accepting or avoiding taste stimuli following a conditioned taste aversion. PMID- 17428455 TI - Acute inhibition of Rho-kinase improves cardiac contractile function in streptozotocin-diabetic rats. AB - OBJECTIVE: The purpose of the present study was to determine whether increased activation of the RhoA/Rho-kinase (ROCK) pathway occurs in diabetic cardiomyopathy and whether acute inhibition of this pathway improves contractile function of the diabetic heart. METHODS: Male Wistar rats were made diabetic with streptozotocin. Twelve to fourteen weeks later, the effects of acute administration of the ROCK inhibitors Y-27632 and H-1152 on cardiac contractile function were measured both in vitro, in isolated working hearts, and in vivo, using echocardiography. Changes in the expression and activity of RhoA, and the effect of ROCK inhibition on changes in the phosphorylation of the downstream target of ROCK, LIM kinase 2, and on actin polymerization in diabetic hearts were also determined. RESULTS: Perfusion of isolated working hearts from diabetic rats with Y-27632 or H-1152 acutely improved left ventricle developed pressure and the rates of contraction and relaxation. Acute administration of H-1152 also significantly improved the percent fraction shortening, an index of left ventricle contractility, in vivo in diabetic rats. The expression and activity of RhoA in cardiomyocytes from diabetic rats were significantly increased, as was the phosphorylation of LIM kinase 2. This was associated with an increase in actin polymerization (the F-actin to G-actin ratio). Both the increase in LIM kinase 2 phosphorylation and actin polymerization were attenuated by ROCK inhibition. CONCLUSIONS: These data suggest that activation of the RhoA/ROCK signaling pathway plays a critical role in the development of diabetic cardiomyopathy, and that ROCK is an excellent therapeutic target in the treatment of this condition. PMID- 17428456 TI - Microsatellite mutation in the maternally/paternally transmitted D18S51 locus: two cases of allele mismatch in the child. AB - BACKGROUND: We investigated 2 cases of paternity dispute with 17 autosomal short tandem repeats (STR), that indicated a mismatch to the maternally and paternally inherited allele at D18S51 locus in children under inquiry. METHODS: 17 autosomal and Y STR loci were analyzed using AmpFlSTR Identifiler, PowerPlex 16, AmpFlSTR(R)Y-filertrade mark kits. The mitochondrial DNA hypervariable regions HV1 and HV2 and 6 STR markers on X chromosome were amplified and sequenced. RESULTS: In case M1, allelic representation in the mother, questioned child and suspected father was 14/19, 12/20 and 12/14 respectively. A complete match with the mother at 6 X STR loci and mitochondrial hypervariable regions was observed. In case F1, allelic representation was 13/14, 14/20 and 16/18 respectively. A complete match with the father at 17 Y chromosome STR loci was observed. D18S51 sequence analysis indicates the expansion of 1 repeat in M1 and 2 repeats in F1 leading to allele mismatch in the child. CONCLUSION: The probability of maternity and paternity were 0.999999 and 0.999999 respectively. This is the first report of a maternally/paternally transmitted D18S51 mutations in the paternity DNA testing. These results conclusively determined that the mother and suspected father are the biological parents of the questioned children in both the cases. PMID- 17428457 TI - Comparison of the CEDIA and MEIA assays for the measurement of sirolimus in organ transplant recipients. AB - OBJECTIVES: This study evaluated two immunoassays, the CEDIA assay and the MEIA assay, used for the measurement of whole blood levels of sirolimus in organ transplant recipients. DESIGN AND METHODS: We report on the performance characteristics (total precision, limit of quantitation (functional sensitivity), limit of detection (analytical sensitivity), linearity, accuracy) for each assay. Patient correlation studies were performed, and the results were analyzed using Bland-Altman plots and Passing-Bablok analysis. RESULTS: Total precision for the MEIA assay, corresponding to three mean concentrations of 5.0, 10.6 and 20.2 ng/mL, was 10.5, 8.5, and 6.7%, respectively. The limit of detection was determined to be 1.1 ng/mL and the limit of quantitation was 1.5 ng/mL. The mean recovery for CEDIA was 105.4%, and analysis of proficiency material demonstrated a large negative bias with respect to the mass spectrometry peer mean-later determined to be due to matrix interference. Results for the CEDIA assay showed a total precision, corresponding to a mean concentration of 5.4, 10.5 and 20.7 ng/mL, of 13.5, 5.6, and 4.1%, respectively. The limit of detection was found to be 4.8 ng/mL, with a limit of quantitation of 5.2 ng/mL. The mean recovery for MEIA was 110.1%, and analysis of proficiency material demonstrated good agreement with the mass spectrometry peer mean with a slight positive bias. Both assays were acceptably linear over the reportable range of the assay. Patient correlation studies demonstrated a positive average bias for both assays versus results from LC-MS measurement (0.9 ng/mL for MEIA, 2.1 ng/mL for CEDIA). CONCLUSION: Based on this evaluation, the MEIA demonstrated acceptable performance for use in clinical monitoring of sirolimus. However, based on a higher limit of quantitation that falls within the therapeutic interval, the CEDIA is not recommended for clinical monitoring of sirolimus. PMID- 17428458 TI - Effect of temperature on protein binding of cortisol. AB - OBJECTIVES: Biological effects of cortisol are substantially determined by protein binding of the hormone. The aim of our study was to characterize temperature effects on cortisol protein binding by use of an equilibrium dialysis method. DESIGN AND METHODS: Serum samples obtained from ten healthy volunteers were submitted to equilibrium dialysis. Each sample from the individuals was incubated for 16 h at 37 degrees C, 38 degrees C, 39 degrees C, 40 degrees C and 41 degrees C, respectively. In the dialysate samples obtained, cortisol concentrations were measured by immunoassay. RESULTS: For samples incubated at 37 degrees C, a mean dialysate cortisol concentration of 0.41 microg/dL (SD=0.14) was found. Gradual increase of dialysate cortisol concentration was observed with increasing incubation temperatures. For samples incubated at 41 degrees C, a mean dialysate cortisol of 0.75 microg/dL (SD=0.24) was found. Thus, the mean percentage of free-to-total cortisol increased by about 80% from 3.7% (SD=1.1) at 37 degrees C to 6.7% (SD=1.8) at 41 degrees C. CONCLUSIONS: The results of our in vitro experiments suggest that during fever the free-to-total ratio of cortisol is increased substantially compared to normal conditions, and that administration of antipyretic drugs might potentially be associated with substantial changes in the bioavailability of cortisol. PMID- 17428459 TI - Gc-globulin concentrations and C5 haplotype-tagging polymorphisms contribute to variations in serum activity of complement factor C5. AB - OBJECTIVES: To investigate the role of Gc-globulin and C5 gene variants as co factors in the regulation of profibrogenic C5 serum activities. DESIGN: Retrospective clinical investigation with 100 healthy probands. Genomic DNA was isolated from whole blood and examined for the human C5 htSNPs rs17611 and rs2300929. Actin-free Gc-globulin-, total Gc-globulin- and total C5 concentrations in serum were measured using ELISA assays; C5 activities in serum were determined using radial immunodiffusion. RESULTS: C5 serum concentrations were significantly elevated in individuals carrying at least one profibrogenic allele of the C5 htSNP rs17611, but no association between C5 htSNPs and C5 serum activities was detected, albeit C5 activities correlated positively with C5 concentrations in serum. However, C5 activities were also positively correlated with total and actin-free Gc-globulin concentrations. CONCLUSION: Our findings indicate that C5 gene variants and Gc-globulin levels co-define the proinflammatory and profibrogenic effects of C5 in patients at-risk for progression of liver fibrosis. PMID- 17428461 TI - Germline stem cells and neo-oogenesis in the adult human ovary. AB - It remains unclear whether neo-oogenesis occurs in postnatal ovaries of mammals, based on studies in mice. We thought to test whether adult human ovaries contain germline stem cells (GSCs) and undergo neo-oogenesis. Rather than using genetic manipulation which is unethical in humans, we took the approach of analyzing the expression of meiotic marker genes and genes for germ cell proliferation, which are required for neo-oogenesis, in adult human ovaries covering an age range from 28 to 53 years old, compared to testis and fetal ovaries served as positive controls. We show that active meiosis, neo-oogenesis and GSCs are unlikely to exist in normal, adult, human ovaries. No early meiotic-specific or oogenesis associated mRNAs for SPO11, PRDM9, SCP1, TERT and NOBOX were detectable in adult human ovaries using RT-PCR, compared to fetal ovary and adult testis controls. These findings are further corroborated by the absence of early meiocytes and proliferating germ cells in adult human ovarian cortex probed with markers for meiosis (SCP3), oogonium (OCT3/4, c-KIT), and cell cycle progression (Ki-67, PCNA), in contrast to fetal ovary controls. If postnatal oogenesis is confirmed in mice, then this species would represent an exception to the rule that neo oogenesis does not occur in adults. PMID- 17428460 TI - The competence of Xenopus blastomeres to produce neural and retinal progeny is repressed by two endo-mesoderm promoting pathways. AB - Only a subset of cleavage stage blastomeres in the Xenopus embryo is competent to contribute cells to the retina; ventral vegetal blastomeres do not form retina even when provided with neuralizing factors or transplanted to the most retinogenic position of the embryo. These results suggest that endogenous maternal factors in the vegetal region repress the ability of blastomeres to form retina. Herein we provide three lines of evidence that two vegetal-enriched maternal factors (VegT, Vg1), which are known to promote endo-mesodermal fates, negatively regulate which cells are competent to express anterior neural and retinal fates. First, both molecules can repress the ability of dorsal-animal retinogenic blastomeres to form retina, converting the lineage from neural/retinal to non-neural ectodermal and endo-mesodermal fates. Second, reducing the endogenous levels of either factor in dorsal-animal retinogenic blastomeres expands expression of neural/retinal genes and enlarges the retina. The dorsal-animal repression of neural/retinal fates by VegT and Vg1 is likely mediated by Sox17alpha and Derriere but not by XNr1. VegT and Vg1 likely exert their effects on neural/retinal fates through at least partially independent pathways because Notch1 can reverse the effects of VegT and Derriere but not those of Vg1 or XNr1. Third, reduction of endogenous VegT and/or Vg1 in ventral vegetal blastomeres can induce a neural fate, but only allows expression of a retinal fate when both BMP and Wnt signaling pathways are concomitantly repressed. PMID- 17428462 TI - Survivin and inducible nitric oxide synthase production during 4NQO-induced rat tongue carcinogenesis: a possible relationship. AB - This study was undertaken to investigate, by immunohistochemistry, the expression of survivin and inducible nitric oxide synthase during 4NQO-induced rat tongue carcinogenesis. Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12, and 20 weeks. Ten animals were used as negative control. Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure, survivin and iNOS were expresssed (p<0.05) in some cells of the 'normal' oral epithelium. In pre-neoplastic lesions at 12 weeks following carcinogen exposure, the levels of survivin and iNOS were increased (p<0.05) when compared to negative control, being the strongest effect observed to iNOS. In well-differentiated squamous cell carcinoma induced after 20 weeks of treatment with 4NQO, survivin and iNOS were expressed in some tumor cells. Lack of immunoreactivity for both markers was observed in the negative control group. Taken together, our results support the belief that expression of survivin and iNOS are early events during malignant transformation and conversion of the oral mucosa. PMID- 17428463 TI - TOR-induced resistance to toxic adenosine analogs in Leishmania brought about by the internalization and degradation of the adenosine permease. AB - TOR is an atypical multidrug resistance protein present in the human protozoan parasite, Leishmania. Resistance to the toxic adenosine analog tubercidin was brought about by redirecting the adenosine permease from the plasma membrane to the multivesicular tubule lysosome. The cells became resistant to tubercidin because they were unable to take up and accumulate this toxic purine. The domain, which was recognized by TOR in this internalization pathway, was identified by expressing portions of this transporter in Leishmania and assessing whether they were capable of hindering the multidrug resistance capability of TOR. This approach identified the adenosine permease region spanning Met289 to Trp305. This region was also the epitope recognized by the internalization mechanism. An internal deletion mutant lacking Met289-Trp305 was functionally active but could no longer be internalized in cells with high TOR levels. The internalization and altered trafficking of the adenosine permease by TOR was observed in yeast and human embryonic kidney cells co-expressing these two Leishmania proteins indicating that the internalization process was conserved in evolutionary diverse organisms. The inability of Saccharomyces with a temperature-sensitive ubiquitin ligase to internalize adenosine permease suggested that ubiquitination was involved in this altered trafficking. PMID- 17428464 TI - Sensitive detection of GFP utilizing tyramide signal amplification to overcome gene silencing. AB - The green fluorescent protein (GFP) is among the most commonly used expression markers in biology. GFP-tagged cells have played a particularly important role in studies of cell lineage. Sensitive detection of GFP is crucially important for such studies to be successful, and problems with detection may account for discrepancies in the literature regarding the possible fate choices of stem cells. Here we describe a very sensitive technique for visualization of GFP. Using it we can detect about 90% of cells of donor origin while we could only see about 50% of these cells when we employ the methods that are in general use in other laboratories. In addition, we provide evidence that some cells permanently silence GFP expression. In the case of the progeny of bone marrow stem cells, it appears that the more distantly related they are to their precursors, the more likely it is that they will turn off the lineage marker. PMID- 17428465 TI - Synthetic nanostructures inducing differentiation of human mesenchymal stem cells into neuronal lineage. AB - Human mesenchymal stem cells (hMSCs) have been shown to trans-differentiate into neuronal-like cells by culture in neuronal induction media, although the mechanism is not well understood. Topography can also influence cellular responses including enhanced differentiation of progenitor cells. As extracellular matrix (ECM) in vivo comprises topography in the nanoscale, we hypothesize that nanotopography could influence stem cell differentiation into specific non-default pathways, such as transdifferentiation of hMSCs. Differentiation and proliferation of hMSCs were studied on nanogratings of 350 nm width. Cytoskeleton and nuclei of hMSCs were aligned and elongated along the nanogratings. Gene profiling and immunostaining showed significant up-regulation of neuronal markers such as microtubule-associated protein 2 (MAP2) compared to unpatterned and micropatterned controls. The combination of nanotopography and biochemical cues such as retinoic acid further enhanced the up-regulation of neuronal marker expressions, but nanotopography showed a stronger effect compared to retinoic acid alone on unpatterned surface. This study demonstrated the significance of nanotopography in directing differentiation of adult stem cells. PMID- 17428466 TI - Phosphatidylinositol 3-kinase/Akt signaling enhances nuclear localization and transcriptional activity of BRCA1. AB - Signaling pathways involved in regulating nuclear-cytoplasmic distribution of BRCA1 have not been previously reported. Here, we provide evidence that heregulin beta1-induced activation of the Akt pathway increases the nuclear content of BRCA1. First, treatment of T47D breast cancer cells with heregulin beta1 results in a two-fold increase in nuclear BRCA1 as assessed by FACS analysis, immunoblotting and immunofluorescence. This heregulin-induced increase in nuclear BRCA1 is blocked by siRNA-mediated down-regulation of Akt. Second, mutation of threonine 509 in BRCA1, the site of Akt phosphorylation, to an alanine, attenuates the ability of heregulin to induce BRCA1 nuclear accumulation. These data suggest that Akt-catalyzed phosphorylation of BRCA1 is required for the heregulin-regulated nuclear concentration of BRCA1. Because most functions ascribed to BRCA1 occur within the nucleus, we postulated that phosphorylation dependent nuclear accumulation of BRCA1 would result in enhanced nuclear activity, specifically transcriptional activity, of BRCA1. This postulate is affirmed by our observation that the ability of BRCA1 to transactivate GADD45 promoter constructs was enhanced in T47D cells treated with heregulin beta1. Furthermore, the heterologous expression of BRCA1 in HCC1937 human breast cancer cells, which have constitutively active Akt, also induces GADD45 promoter activity, whereas the expression of BRCA1 in which threonine 509 has been mutated to an alanine is able to only minimally induce promoter activity. These findings implicate Akt in upstream events leading to BRCA1 nuclear localization and function. PMID- 17428467 TI - Troglitazone induces differentiation in Trypanosoma brucei. AB - Trypanosoma brucei, a protozoan parasite causing sleeping sickness, is transmitted by the tsetse fly and undergoes a complex lifecycle including several defined stages within the insect vector and its mammalian host. In the latter, differentiation from the long slender to the short stumpy form is induced by a yet unknown factor of trypanosomal origin. Here we describe that some thiazolidinediones are also able to induce differentiation. In higher eukaryotes, thiazolidinediones are involved in metabolism and differentiation processes mainly by binding to the intracellular receptor peroxisome proliferator activated receptor gamma. Our studies focus on the effects of troglitazone on bloodstream form trypanosomes. Differentiation was monitored using mitochondrial markers (membrane potential, succinate dehydrogenase activity, inhibition of oxygen uptake by KCN, amount of cytochrome transcripts), morphological changes (Transmission EM and light microscopy), and transformation experiments (loss of the Variant Surface Glycoprotein coat and increase of dihydroliponamide dehydrogenase activity). To further investigate the mechanisms responsible for these changes, microarray analyses were performed, showing an upregulation of expression site associated gene 8 (ESAG8), a potential differentiation regulator. PMID- 17428468 TI - Kinetic analyses as a critical parameter in defining the side population (SP) phenotype. AB - The side population (SP) phenotype has been reported as a method to identify hematopoietic stem cells in the bone marrow based upon differential staining with the fluorescent dye, Hoechst 33342. This technique has drawn great interest in the stem cell community, as it may provide a simple approach to the enrichment of progenitor cells from a variety of normal and malignant tissues. The frequency of these cells and their performance in functional assays has varied considerably within the literature. To investigate mechanisms that may contribute to the SP phenotype, we measured the fluorescence emission of Hoechst-stained bone marrow cells as a function of both time and dye concentration using a custom flow cytometer and data acquisition software. These measurements demonstrate that all nucleated cells within the bone marrow undergo an identical staining pattern at varying rates, even under conditions previously reported to abrogate the SP. Therefore, the SP phenotype is not unique to stem cells, but rather represents a transient feature of marrow cells exposed to Hoechst 33342 for varying amounts of time. We propose that heterogeneity of SP-defined populations may be a consequence of the rate at which differing cell populations accumulate Hoechst 33342. Further, we suggest that dye uptake kinetics will likely be an important factor for optimal use of Hoechst 33342 in isolating stem cells. PMID- 17428469 TI - Regulation of transient receptor potential channels of melastatin type 8 (TRPM8): effect of cAMP, cannabinoid CB(1) receptors and endovanilloids. AB - The transient receptor potential channel of melastatin type 8 (TRPM8), which is gated by low (<25 degrees C) temperature and chemical compounds, is regulated by protein kinase C-mediated phosphorylation in a way opposite to that observed with the transient receptor potential channel of vanilloid type 1 (TRPV1), i.e. by being desensitized and not sensitized. As TRPV1 is sensitized also by protein kinase A (PKA)-mediated phosphorylation, we investigated the effect of two activators of the PKA pathway, 8-Br-cAMP and forskolin, on the activity of menthol and icilin at TRPM8 in HEK-293 cells stably overexpressing the channel (TRPM8-HEK-293 cells). We also studied the effect on TRPM8 of: (1) a series of compounds previously shown to activate or antagonize TRPV1, and (2) co stimulation of transiently co-expressed cannabinoid CB(1) receptors. Both 8-Br cAMP (100 microM) and forskolin (10 microM) right-shifted the dose-response curves for the TRPM8-mediated effect of icilin and menthol on intracellular Ca(2+). The inhibitory effects of 8-Br-cAMP and forskolin were attenuated by the selective PKA inhibitor Rp-cAMP-S. Stimulation of human CB(1) receptors transiently co-expressed in TRPM8-HEK-293 cells also inhibited TRPM8 response to icilin. Finally, some TRPV1 agonists and antagonists, but not iodinated antagonists, antagonized icilin- and much less so menthol-, induced TRPM8 activation. Importantly, the endovanilloids/endocannabinoids, anandamide and NADA, also antagonized TRPM8 at submicromolar concentrations. Although these findings need to be confirmed by experiments directly measuring TRPM8 activity in natively TRPM8-expressing cells, they support the notion that the same regulatory events have opposing actions on TRPM8 and TRPV1 receptors and identify anandamide and NADA as the first potential endogenous functional antagonists of TRPM8 channels. PMID- 17428470 TI - Hair follicle-specific keratins and their diseases. AB - The human keratin family comprises 54 members, 28 type I and 26 type II. Out of the 28 type I keratins, 17 are epithelial and 11 are hair keratins. Similarly, the 26 type II members comprise 20 epithelial and 6 hair keratins. As, however, 9 out of the 37 epithelial keratins are specifically expressed in the hair follicle, the total number of hair follicle-specific keratins (26) almost equals that of those expressed in the various forms of epithelia (28). Up to now, more than half of the latter have been found to be involved in inherited diseases, with mutated type I and type II members being roughly equally causal. In contrast, out of the 26 hair follicle-specific keratins only 5 have, at present, been associated with inherited hair disorders, while one keratin merely acts as a risk factor. In addition, all hair follicle-specific keratins involved in pathologies are type II keratins. Here we provide a detailed description of the respective hair diseases which are either due to mutations in hair keratins (monilethrix, ectodermal dysplasia of hair and nail type) or hair follicle specific epithelial keratins (two mouse models, RCO3 and Ca(Rin) as well as pseudofolliculitis barbae). PMID- 17428471 TI - Gadd45a, the gene induced by the mood stabilizer valproic acid, regulates neurite outgrowth through JNK and the substrate paxillin in N1E-115 neuroblastoma cells. AB - Valproic acid (VPA), a mood stabilizer and anticonvulsant, has a variety of neurotrophic functions; however, less is known about how VPA regulates neurite outgrowth. Here, using N1E-115 neuroblastoma cells as the model, we show that VPA upregulates Gadd45a to trigger activation of the downstream JNK cascade controlling neurite outgrowth. VPA induces the phosphorylation of c-Jun N terminal kinase (JNK) and the substrate paxillin, while VPA induction of neurite outgrowth is inhibited by JNK inhibitors (SP600125 and the small JNK-binding peptide) or a paxillin construct harboring a Ser 178-to-Ala mutation at the JNK phosphorylation. Transfection of Gadd45a, acting through the effector MEKK4, leads to the phosphorylation of the JNK cascade. Conversely, knockdown of Gadd45a with siRNA reduces the effect of VPA. Taken together, these results suggest that upregulation of Gadd45a explains one of the mechanisms whereby VPA induces the neurotrophic effect, providing a new role of Gadd45a in neurite outgrowth. PMID- 17428472 TI - High tolerance to apoptotic stimuli induced by serum depletion and ceramide in side-population cells: high expression of CD55 as a novel character for side population. AB - Cancer stem cells are supposed to be resistant to apoptosis, but information for this is quite limited. Cancer stem cells are usually isolated as dye-effluxing cells with Hoechst 33342 staining, called side-population (SP) cells. Because Hoechst 33342 dye itself induces apoptosis, the SP cells isolated by such method are not suitable for evaluation of apoptosis. For accurate assessment, SP cells must be isolated without Hoechst 33342. Here, we found that CD55 was highly expressed in SP cells of two mammary gland carcinoma cell lines. Then, the high expression of CD55 was used for isolation of cancer stem cells among mammary carcinoma cell lines as a surrogate character. Cells expressing high level of CD55 (CD55(hi)) were resistant to apoptosis induced by serum depletion as in the case of SP cells. In ceramide-inducing apoptosis, CD55(hi) cells showed high tolerance. Anti-apoptotic molecules such as Bcl-2 were abundantly expressed in both SP cells and CD55(hi) cells. These findings indicated that SP cells as revealed to be CD55(hi) cells were tolerant to apoptosis. The high expression of CD55 may be a useful character for SP cells in evaluating their functions. PMID- 17428473 TI - Post-transcriptional control of neurofilaments in development and disease. AB - Tight coordination of the expression of neurofilament subunits is integral to the normal development and function of the nervous system. Imbalances in their expression are increasingly implicated in the induction of neurodegeneration in which formation of neurofilamentous aggregates is central to the pathology. Neurofilament expression can be controlled not only at the transcriptional level but also through post-transcriptional regulation of mRNA localization, stability, and translational efficiency. The critical role that post-transcriptional mechanisms play in maintaining neurofilament homeostasis is highlighted, for example, by the human disease amyotrophic lateral sclerosis, in which selective destabilization of NF-L mRNA (or failure to stabilize it) is associated with the formation of neurofilamentous aggregates - a hallmark of the disease process. This review discusses the post-transcriptional regulatory mechanisms and associated ribonucleoproteins that have been implicated to date in controlling neurofilament expression during normal development and in disrupting neurofilament homeostasis during neurodegenerative disease. PMID- 17428474 TI - Electrical stimulation promotes sensory neuron regeneration and growth-associated gene expression. AB - Brief electrical stimulation enhances the regenerative ability of axotomized motor [Nix, W.A., Hopf, H.C., 1983. Electrical stimulation of regenerating nerve and its effect on motor recovery. Brain Res. 272, 21-25; Al-Majed, A.A., Neumann, C.M., Brushart, T.M., Gordon, T., 2000. Brief electrical stimulation promotes the speed and accuracy of motor axonal regeneration. J. Neurosci. 20, 2602-2608] and sensory [Brushart, T.M., Jari, R., Verge, V., Rohde, C., Gordon, T., 2005. Electrical stimulation restores the specificity of sensory axon regeneration. Exp. Neurol. 194, 221-229] neurons. Here we examined the parameter of duration of stimulation on regenerative capacity, including the intrinsic growth programs, of sensory neurons. The effect of 20 Hz continuous electrical stimulation on the number of DRG sensory neurons that regenerate their axons was evaluated following transection and surgical repair of the femoral nerve trunk. Stimulation was applied proximal to the repair site for 1 h, 3 h, 1 day, 7 days or 14 days at the time of nerve repair. Following a 21-day regeneration period, DRG neurons that regenerated axons into the muscle and cutaneous sensory nerve branches were retrogradely identified. Stimulation of 1 h led to a significant increase in DRG neurons regenerating into cutaneous and muscle branches when compared to 0 h (sham) stimulation or longer periods of stimulation. Stimulation for 1 h also significantly increased the numbers of neurons that regenerated axons beyond the repair site 4 days after lesion and was correlated with a significant increase in expression of growth-associated protein 43 (GAP-43) mRNA in the regenerating neurons at 2 days post-repair. An additional indicator of heightened plasticity following 1 h stimulation was elevated expression of brain-derived neurotrophic factor (BDNF). The effect of brief stimulation on enhancing sensory and motoneuron regeneration holds promise for inducing improved peripheral nerve repair in the clinical setting. PMID- 17428475 TI - Abundance of microRNA target motifs in the 3'-UTRs of 20527 human genes. AB - A mechanism, selective avoidance, proposes that microRNA (miRNA) target sites are selectively depleted in the 3'-UTRs of genes expressed at the same time and place as a miRNA. If this mechanism is ubiquitous, the target motif occurrences in the 3'-UTRs would be decreased. To test this hypothesis, we examined miRNA target motif occurrences in the 3'- and 5'-UTRs of 20527 human protein-coding genes. The results revealed that miRNA target motifs appeared more frequently than non target motifs and were enriched in the 3'-UTRs. This enrichment was relatively reduced in a set of 2525 genes coexpressed with miR-124a in the prefrontal cortex, but still remained at a high level, suggesting that miRNA target motifs are fostered by some other factors that surpass the influence of selective avoidance. PMID- 17428476 TI - Stress molecules in sepsis and systemic inflammatory response syndrome. AB - During sepsis, microbial derived products ("pathogen-associated molecular patterns", PAMPs) are recognized as exogenous danger signals by specific sensors of the host ("pattern recognitions receptors", PRRs). This interaction leads to the release of numerous stress proteins that are a prerequisite to fight infection, though their overzealous production can contribute to tissue damage, organ dysfunction and eventually death. In critically ill patients, translocation of PAMPs can occur from the gut, and injured tissues and cells release endogenous danger signals called "alarmins" (e.g. High mobility group box-1); that share some properties with PAMPs. Thus, numerous similarities occur during infectious and non-infectious systemic inflammation. PMID- 17428477 TI - GM1 and GM3 gangliosides highlight distinct lipid microdomains within the apical domain of epithelial cells. AB - The apical domain of epithelial cells is composed of distinct subdomains such as microvilli, primary cilia and a non-protruding region. Using the cholesterol binding protein prominin-1 as a specific marker of plasma membrane protrusions we have previously proposed the co-existence of different cholesterol-based lipid microdomains (lipid rafts) within the apical domain [Roper, K., Corbeil, D. and Huttner, W.B. (2000), Retention of prominin in microvilli reveals distinct cholesterol-based lipid microdomains in the apical plasma membrane. Nat. Cell Biol. 2, 582-592]. To substantiate the hypothesis that the microvillar plasma membrane subdomains contain a distinct set of lipids compared to the planar portion we have investigated the distribution of prominin-1 and two raft associated gangliosides GM(1) and GM(3) by fluorescence microscopy. GM(1) was found to co-localize with prominin-1 on microvilli whereas GM(3) was segregated from there suggesting its localization in the planar region. Regarding the primary cilium, overlapping fluorescent signals of GM(1) or GM(3) and prominin-1 were observed. Thus, our data demonstrate that specific ganglioside-enriched rafts are found in different apical subdomains and reveal that two plasma membrane protrusions with different structural bases (actin for the microvillus and tubulin for the cilium) are composed of distinct types of lipid. PMID- 17428478 TI - Pelvic pain after gonadotropin administration as a potential sign of endometriosis. AB - We describe five patients who developed significant pelvic pain, requiring narcotics, during a controlled ovarian hyperstimulation cycle and who were surgically diagnosed with significant endometriosis. Severe pain, especially if it requires narcotics, is unusual for patients undergoing controlled ovarian hyperstimulation and may be an indicator of endometriosis. PMID- 17428479 TI - GnRH agonist and antagonist protocols for stage I-II endometriosis and endometrioma in in vitro fertilization/intracytoplasmic sperm injection cycles. AB - OBJECTIVE: To investigate the outcomes of intracytoplasmic sperm injection (ICSI) cycles after controlled ovarian hyperstimulation (COH) with GnRH antagonist or GnRH agonist (GnRH-a) in mild-to-moderate endometriosis and endometrioma. DESIGN: Prospective randomize trial. SETTING: A private IVF center. PATIENT(S): A total of 246 ICSI cycles in 246 patients were divided into three groups: women with mild-to-moderate endometriosis (n = 98); women who had ovarian surgery for endometrioma (n = 81); women with endometrioma and no history of previous surgery (n = 67). INTERVENTION(S): Patients in each group were randomized to COH with either triptrolein or cetrorelix. MAIN OUTCOME MEASURE(S): Clinical parameters, characteristics of COH, and ICSI results were analyzed. RESULT(S): Outcomes of COH with both GnRH antagonist and GnRH-a were similar in patients with mild-to moderate endometriosis. Implantation rates were 15.9% vs. 22.6% and clinical pregnancy rates were 27.5% vs. 39% with GnRH antagonist and GnRH-a protocols, respectively, in patients who had ovarian surgery for endometrioma. Implantation rates were 12.5% vs. 14.8% and clinical pregnancy rates were 20.5% vs. 24.2% with GnRH antagonist and GnRH-a protocols, respectively, in patients with endometrioma and no history of ovarian surgery. CONCLUSION(S): Considering the implantation and clinical pregnancy rates, COH with both GnRH antagonist and GnRH-a protocols may be equally effective in patients with mild-to-moderate endometriosis and endometrioma who did and did not undergo ovarian surgery. PMID- 17428480 TI - Letrozole for ovulation induction and fertility preservation by embryo cryopreservation in young women with endometrial carcinoma. AB - OBJECTIVE: To reduce estrogen exposure in women with endometrial cancer undergoing in vitro fertilization using an aromatase inhibitor. DESIGN: Prospective case series. SETTING: Academic center for reproductive medicine. PATIENT(S): Endometrial carcinoma patients presenting for fertility preservation or fresh embryo transfer to gestational carrier. INTERVENTION(S): Four patients with endometroid carcinoma underwent five IVF cycles for immediate or delayed embryo transfer to gestational carriers before or after staging and definitive surgery. To prevent surge in E(2) levels, letrozole was started 2 days before gonadotropin administration and then given concomitantly. Embryos were either cryopreserved for fertility preservation or transferred freshly to a surrogate. MAIN OUTCOME MEASURE(S): Peak E(2) level during stimulation, pregnancy in a gestational carrier. RESULT(S): Peak E(2) level during stimulation was 386.67 +/- 102.93 pg/mL. A mean of 7 +/- 2.85 oocytes were retrieved, resulting in 4.8 +/- 1.76 embryos per cycle. In one patient, two embryos were transferred to a surrogate, resulting in a triplet pregnancy. The delivery occurred at 31 weeks and the infants did not show any congenital or developmental abnormalities. Three patients had their embryos cryopreserved for future use. CONCLUSION(S): The use of letrozole and gonadotropins is associated with lower E(2) levels compared with standard stimulation cycles in endometrial carcinoma patients. Combination of this approach with surrogacy may enable these young women to preserve their fertility. PMID- 17428481 TI - Clinical Challenges and Images in GI. Solid pseudopapillary tumor of the pancreas. PMID- 17428482 TI - Predicting surgical risk in patients with cirrhosis: from art to science. PMID- 17428483 TI - Characterization and quantification of corticosteroid-binding globulin in a southern toad, Bufo terrestris, exposed to coal-combustion-waste. AB - Corticosteroid-binding globulin (CBG) is a plasma protein that binds corticosterone and may regulate access of hormone to tissues. The role of CBG during a stress response is not clear. At least two hypotheses have been proposed: 1) CBG levels may increase in response to a stressor, thereby decreasing the amount of circulating free corticosterone, or 2) CBG levels may decline, making corticosterone available for its role in increased metabolic needs during stress. In this study, southern toads, Bufo terrestris, were exposed to a chronic pollutant (coal-combustion-waste), to determine changes in CBG and free corticosterone levels. Since toads exposed to chronic pollutants in previous studies did not exhibit the predicted changes in metabolic rate and mass, but did experience a significant elevation in total corticosterone, we hypothesized that CBG would likewise increase and thus, mitigate the effects of a chronic (i.e. 2 months) pollutant stressor. To conduct this study, we first characterized the properties of CBG in southern toads. Toad CBG has a K(d)=20.6+/-1.0 nM and a B(max)=332.2+/-5.1 nmol/L plasma. The rank order potencies for steroid inhibition of tritiated corticosterone are: dihydrotestosterone > corticosterone >> progesterone=testosterone >>> estrogen=dexamethasone. After characterization, we monitored the changes in CBG, total corticosterone, and free corticosterone in male toads that were exposed to either coal-combustion-waste or control conditions. CBG increased in all groups throughout the experiment. Total corticosterone, on the other hand, was only significantly elevated at four weeks of exposure to coal-combustion-waste. The increase in CBG did not parallel the increase in total corticosterone; as a result, free corticosterone levels were not buffered by CBG, but showed a peak at four weeks similar to total corticosterone. This finding indicates that, in this species, CBG may not provide a protective mechanism during long-term pollution exposure. PMID- 17428484 TI - Intraperitoneal delivery of exogenous corticosterone via osmotic pump in a passerine bird. AB - Selecting the most reliable method to deliver exogenous steroids remains a problem for researchers, particularly when designing experiments on small birds. We used intraperitoneal (IP) osmotic pumps to deliver exogenous corticosterone (CORT) and RU486 in captive White-throated Sparrows. Males received implants containing either a low (LD) or moderate dose (MD) of CORT, RU486 (RU), or polyethylene glycol vehicle only (V). This method provided sustained elevations in baseline CORT in both LD and MD males compared to V males, with higher CORT levels induced in MD males. Slight increases in post-implant CORT resulted in V males, but not in RU males. We observed no significant change in the condition of V males in terms of body mass, furcular fat score or food intake rates, although locomotor activity declined slightly after implantation. Taken together, our results suggest that IP pumps had little impact on the overall health of the birds. CORT levels were maintained within natural ranges for this species, suggesting that our results are biologically relevant and useful for future endocrine studies with small birds. PMID- 17428485 TI - Developmental exposure to xenoestrogen enhances spatial learning in male rats. AB - Steroid hormones have profound effects on the development and function of the nervous system. Environmental estrogens or xenoestrogens are manmade or are natural compounds, which mimics the action of estrogen hormones. The experimental evidence for impairment of cognitive functions in humans and mammals following exposure to xenoestrogens has been fiercely debated. The strongest arguments against such studies have been that the route, time course, and intensity of exposure did not simulate environmental exposure, and that the chemicals tested have additional, non-estrogenic toxic effects, hindering a generalization of actual "xenoestrogenic" effects. Here we show that an environmental-like exposure to the pure estrogen, 17alpha-ethynylestradiol (EE2) during development enhances spatial learning abilities in adult male Sprague-Dawley rats. To simulate an environmental exposure, we used a very low dose (4 ng/kg/day) of EE2 equivalent to concentrations measured in European and US streams which was given orally with a non-invasive method, and we extended the treatment for the entire course of development, from conception to puberty. The animals were tested in a Morris water maze protocol at 6 months of age. Male rats treated with EE2 during development showed a faster learning during the training phase, and remembered better the position of the hidden platform in the short term. Our study demonstrates that actual levels of exposure to xenoestrogens can permanently alter cognitive abilities of a mammalian species. PMID- 17428486 TI - Finasteride, a 5alpha-reductase inhibitor, potentiates antinociceptive effects of morphine, prevents the development of morphine tolerance and attenuates abstinence behavior in the rat. AB - It has been shown that morphine increases 5alpha-reductase enzyme activity in the rat central nervous system; however importance of this finding on morphine analgesia, tolerance and dependence has not been reported. In the present study, we investigated inhibition of 5alpha-reductase enzyme on morphine effects using finasteride. To determine whether the 5alpha-reductase enzyme interact with morphine analgesia, finasteride (5 mg/kg, i.p.) was administrated with morphine (5 and 7 mg/kg, i.p.). The tail-flick test was used to assess the nociceptive threshold, before and 15, 30, 45, 60 and 90 min after drug administration. In tolerance experiments, morphine 20 mg/kg was injected i.p., twice daily for 4 days. The development and expression of dependence were assessed in the naloxone precipitation test 5 days after the morphine (20-30 mg/kg, i.p.) administration. We found that finasteride could potentiate the antinociceptive effect of morphine. In addition, chronic finasteride administration effectively blocked development of tolerance and dependence to morphine. Following chronic morphine administration, single dose injection of finasteride failed to reverse tolerance but prevented naloxone precipitate withdrawal syndrome. Therefore, it was concluded that there is a functional relationship between 5alpha-reductase enzyme and morphine. PMID- 17428487 TI - Extension of IMC tuning correlations for non-self regulating (integrating) processes. AB - The filter term of a PID with Filter controller reduces the impact of measurement noise on the derivative action of the controller. This impact is quantified by the controller output travel defined as the total movement of the controller output per unit time. Decreasing controller output travel is important to reduce wear in the final control element. Internal Model Control (IMC) tuning correlations are widely published for PI, PID, and PID with Filter controllers for self regulating processes. For non-self regulating (or integrating) processes, IMC tuning correlations are published for PI and PID controllers but not for PID with Filter controllers. The important contribution of this work is that it completes the set of IMC tuning correlations with an extension to the PID with Filter controller for non-self regulating processes. Other published correlations (not based upon the IMC framework) for PID with Filter controllers fix the filter time constant at one-tenth the derivative time regardless of the model of the process. In contrast, the novel IMC correlations presented in this paper calculate a filter time constant based upon the model of the process and the user's choice for the closed-loop time constant. The set point tracking and disturbance rejection performance of the proposed IMC tunings is demonstrated using simulation studies and a bench-scale experimental system. The proposed IMC tunings are shown to perform as well as various PID correlations (with and without a filter term) while requiring considerably less controller action. PMID- 17428488 TI - Re-assessing the locations of components of the classical vesicle-mediated trafficking machinery in transfected Plasmodium falciparum. AB - The malaria parasite, Plasmodium falciparum, exports proteins beyond the confines of its own plasma membrane, however there is debate regarding the machinery used for these trafficking events. We have generated transgenic parasites expressing chimeric proteins and used immunofluorescence studies to determine the locations of plasmodial homologues of the COPII component, Sar1p, and the Golgi-docking protein, Bet3p. The P. falciparum Sar1p (PfSar1p) chimeras bind to the endoplasmic reticulum surface and define a network of membranes wrapped around parasite nuclei. As the parasite matures, the endomembrane systems of individual merozoites remain interconnected until very late in schizogony. Antibodies raised against plasmodial Bet3p recognise two foci of reactivity in early parasite stages that increase in number as the parasite matures. Some of the P. falciparum Bet3p (PfBet3p) compartments are juxtaposed to compartments defined by the cis Golgi marker, PfGRASP, while others are distributed through the cytoplasm. The compartments defined by the trans Golgi marker, PfRab6, are separate, suggesting that the Golgi is dispersed. Bet3p-green fluorescent protein (GFP) is partly associated with punctate structures but a substantial population diffuses freely in the parasite cytoplasm. By contrast, yeast Bet3p is very tightly associated with immobile structures. This study challenges the view that the COPII complex and the Golgi apparatus are exported into the infected erythrocyte cytoplasm. PMID- 17428489 TI - Purification of the seven tetranortriterpenoids in neem (Azadirachta indica) seed by counter-current chromatography sequentially followed by isocratic preparative reversed-phase high-performance liquid chromatography. AB - Counter-current chromatography (CCC) sequentially followed by isocratic preparative reversed-phase high-performance liquid chromatography was used to isolate the seven bio-actives (azadirachtin A, azadirachtin B, azadirachtin H, desacetylnimbin, desacetylsalannin, nimbin and salannin) from the seed concentrate (NSC) of the neem tree (Azadirachta indica A. Juss). Reproducible, narrow polarity range, high purity fractions were obtained from repeated injections of the NSC (700 mg loadings/injection), on to a relatively small volume CCC coil (116 mL). The CCC biphasic solvent system chosen was hexane:butanol:methanol:water (1:0.9:1:0.9, v/v). A mass balance of injected material showed that 95+% were recovered. PMID- 17428490 TI - Service life of counter-current chromatography coils. AB - A multilayer coil of PTFE tubing, which failed after being used each workday for about 3 years in a type J centrifuge, was examined. Two types of defects were found. One, called crazes, occurs throughout the coil and does not leak initially, but may eventually lead to a short, axially oriented slit. Another, called indentations, is seen primarily in the innermost and other nearby layers. They are elongated, about 5 mm, indentations, usually on the central side of the tubing. These eventually crack and leak. PTFE tubing is permeable to air and hexane and expands by more than 1% when immersed in hexane, heptane or chloroform for a few days. It is suggested that the crazes result from exposure of the somewhat flexible tubing to the undulating centripetal force field in the coil planet centrifuge, especially when further softened by solvent absorption. The indentations may result from carriage of the excess tubing length, created by solvent absorption, from the coil periphery to the coil center by the centripetal force field, which continuously travels from the peripheral tail to the central head of the coil. A 1% increase in coil length creates 74 cm of excess tubing in the 160-ml coils examined in this study. It is suggested that fluorinated ethylene propylene (FEP) tubing, especially when etched on the outside, may provide more stable CCC coils, since its expansion when exposed to organic solvents is 0.1 or less than that of PTFE. PMID- 17428491 TI - Optimization of a rapid microwave assisted extraction method for the liquid chromatography-electrospray-tandem mass spectrometry determination of isoflavonoid aglycones in soybeans. AB - A very fast chromatographic separation of isoflavonoids genistein, daidzein, formononetin and biochanin A was developed on a C18 high-speed column under isocratic conditions. The method was validated in terms of detection limits, quantitation limits (LOQs), linearity and precision. LOQs in 0.04-0.2 microg/g range were calculated, making feasible the determination of these compounds of nutritional concern at trace levels. Good linearity was demonstrated over three concentration orders of magnitude for each analyte (r2 0.990-1.000). The intra day and inter-day repeatability was evaluated in terms of relative standard deviation (RSD%) at two concentration levels for each analyte (RSD% <9%). An optimization strategy was adopted to find the best conditions for the extraction of isoflavonoid aglycones from yellow soybeans using microwave-assisted extraction. The most relevant parameters resulted to be the microwave power, the extraction time and the acid concentration, optimal values being 600 W, 1 min and 12 M, respectively. When performing sample treatment on a fortified soybean sample, high recovery percentage was obtained for both compounds (94+/-8% for daidzein and 97+/-5% (n = 4) for genistein). The concentration level at which daidzein and genistein were found in the soybean sample were 1.21+/-0.15 mg/g and 2.38+/-0.09 mg/g (n=4), respectively. PMID- 17428492 TI - Recent advances in sheathless interfacing of capillary electrophoresis and electrospray ionization mass spectrometry. AB - On line sheathless capillary electrophoresis (CE)-electrospray ionization (ESI) mass spectrometry is developing as a powerful method in bioanalytics as it provides high resolution, sensitivity, relatively short analysis times, and amenability to a wide class of compounds. However, unlike the popular nano liquid chromatography (nano LC) or sheath-flow CE/ESI-MS, the sheathless coupling lacks standardized designs and protocols. For this reason, sheathless CE/ESI is a subject of conceptual and technical upgrading more than any other liquid-based separation method hyphenated to MS. Here, recent innovations in sheathless CE/ESI MS interfacing are gathered in a survey covering the 2005/2006 period. In the first part of the review, the current concepts and methods for in-laboratory production of sturdy designs based on either conductive emitters or electrodeless interfaces are described. The second part is dedicated to microchip CE platforms with externally connected emitters for sheathless coupling to ESI-MS and advanced microfluidic devices integrating CE and sheathless electrospray in a single chip substrate. The advantages, limitations and feasibility for certain applications of all these systems as well as the perspectives for their performance improvement are concurrently assessed. PMID- 17428493 TI - Widespread dispersal of the microsporidian Nosema ceranae, an emergent pathogen of the western honey bee, Apis mellifera. AB - The economically most important honey bee species, Apis mellifera, was formerly considered to be parasitized by one microsporidian, Nosema apis. Recently, [Higes, M., Martin, R., Meana, A., 2006. Nosema ceranae, a new microsporidian parasite in honeybees in Europe, J. Invertebr. Pathol. 92, 93-95] and [Huang, W. F., Jiang, J.-H., Chen, Y.-W., Wang, C.-H., 2007. A Nosema ceranae isolate from the honeybee Apis mellifera. Apidologie 38, 30-37] used 16S (SSU) rRNA gene sequences to demonstrate the presence of Nosema ceranae in A. mellifera from Spain and Taiwan, respectively. We developed a rapid method to differentiate between N. apis and N. ceranae based on PCR-RFLPs of partial SSU rRNA. The reliability of the method was confirmed by sequencing 29 isolates from across the world (N =9 isolates gave N. apis RFLPs and sequences, N =20 isolates gave N. ceranae RFLPs and sequences; 100% correct classification). We then employed the method to analyze N =115 isolates from across the world. Our data, combined with N =36 additional published sequences demonstrate that (i) N. ceranae most likely jumped host to A. mellifera, probably within the last decade, (ii) that host colonies and individuals may be co-infected by both microsporidia species, and that (iii) N. ceranae is now a parasite of A. mellifera across most of the world. The rapid, long-distance dispersal of N. ceranae is likely due to transport of infected honey bees by commercial or hobbyist beekeepers. We discuss the implications of this emergent pathogen for worldwide beekeeping. PMID- 17428494 TI - FtsZ polymer-bundling by the Escherichia coli ZapA orthologue, YgfE, involves a conformational change in bound GTP. AB - Cell division is a fundamental process for both eukaryotic and prokaryotic cells. In bacteria, cell division is driven by a dynamic, ring-shaped, cytoskeletal element (the Z-ring) made up of polymers of the tubulin-like protein FtsZ. It is thought that lateral associations between FtsZ polymers are important for function of the Z-ring in vivo, and that these interactions are regulated by accessory cell division proteins such as ZipA, EzrA and ZapA. We demonstrate that the putative Escherichia coli ZapA orthologue, YgfE, exists in a dimer/tetramer equilibrium in solution, binds to FtsZ polymers, strongly promotes FtsZ polymer bundling and is a potent inhibitor of the FtsZ GTPase activity. We use linear dichroism, a technique that allows structure analysis of molecules within linear polymers, to reveal a specific conformational change in GTP bound to FtsZ polymers, upon bundling by YgfE. We show that the consequences of FtsZ polymer bundling by YgfE and divalent cations are very similar in terms of GTPase activity, bundle morphology and GTP orientation and therefore propose that this conformational change in bound GTP reveals a general mechanism of FtsZ bundling. PMID- 17428495 TI - Structural and functional differences of SWIRM domain subtypes. AB - SWIRM is a conserved domain found in several chromatin-associated proteins. Based on their sequences, the SWIRM family members can be classified into three subfamilies, which are represented by Swi3, LSD1, and Ada2. Here we report the SWIRM structure of human MYb-like, Swirm and Mpn domain-containing protein-1 (MYSM1). The MYSM1 SWIRM structure forms a compact HTH-related fold comprising five alpha-helices, which best resembles the Swi3 SWIRM structure, among the known SWIRM structures. The MYSM1 and Swi3 SWIRM structures are more similar to the LSD1 structure than the Ada2alpha structure. The SWIRM domains of MYSM1 and LSD1 lacked DNA binding activity, while those of Ada2alpha and the human Swi3 counterpart, SMARCC2, bound DNA. The dissimilarity in the DNA-binding ability of the MYSM1 and SMARCC2 SWIRM domains might be due to a couple of amino acid differences in the last helix. These results indicate that the SWIRM family has indeed diverged into three structural subfamilies (Swi3/MYSM1, LSD1, and Ada2 types), and that the Swi3/MYSM1-type subfamily has further diverged into two functionally distinct groups. We also solved the structure of the SANT domain of MYSM1, and demonstrated that it bound DNA with a similar mode to that of the c Myb DNA-binding domain. PMID- 17428496 TI - Solution structure of the MID1 B-box2 CHC(D/C)C(2)H(2) zinc-binding domain: insights into an evolutionarily conserved RING fold. AB - The B-box type 2 domain is a prominent feature of a large and growing family of RING, B-box, coiled-coil (RBCC) domain-containing proteins and is also present in more than 1500 additional proteins. Most proteins usually contain a single B-box2 domain, although some proteins contain tandem domains consisting of both type 1 and type 2 B-boxes, which actually share little sequence similarity. Recently, we determined the solution structure of B-box1 from MID1, a putative E3 ubiquitin ligase that is mutated in X-linked Opitz G/BBB syndrome, and showed that it adopted a betabetaalpha RING-like fold. Here, we report the tertiary structure of the B-box2 (CHC(D/C)C(2)H(2)) domain from MID1 using multidimensional NMR spectroscopy. This MID1 B-box2 domain consists of a short alpha-helix and a structured loop with two short anti-parallel beta-strands and adopts a tertiary structure similar to the B-box1 and RING structures, even though there is minimal primary sequence similarity between these domains. By mutagenesis, ESI-FTICR and ICP mass spectrometry, we show that the B-box2 domain coordinates two zinc atoms with a 'cross-brace' pattern: one by Cys175, His178, Cys195 and Cys198 and the other by Cys187, Asp190, His204, and His207. Interestingly, this is the first case that an aspartic acid is involved in zinc atom coordination in a zinc-finger domain, although aspartic acid has been shown to coordinate non-catalytic zinc in matrix metalloproteinases. In addition, the finding of a Cys195Phe substitution identified in a patient with X-linked Opitz GBBB syndrome supports the importance of proper zinc coordination for the function of the MID1 B-box2 domain. Notably, however, our structure differs from the only other published B-box2 structure, that from XNF7, which was shown to coordinate one zinc atom. Finally, the similarity in tertiary structures of the B-box2, B-box1 and RING domains suggests these domains have evolved from a common ancestor. PMID- 17428497 TI - An ATP hydrolysis sensor in the DNA packaging motor from bacteriophage T4 suggests an inchworm-type translocation mechanism. AB - Tailed bacteriophages and large eukaryotic viruses employ powerful molecular motors to translocate dsDNA into a preassembled capsid shell. The phage T4 motor is composed of a dodecameric portal and small and large terminase subunits assembled at the special head-tail connector vertex of the prohead. The motor pumps DNA through the portal channel, utilizing ATP hydrolysis energy provided by an ATPase present in the large terminase subunit. We report that the ATPase motors of terminases, helicases, translocating restriction enzymes, and protein translocases possess a common coupling motif (C-motif). Mutations in the phage T4 terminase C-motif lead to loss of stimulated ATPase and DNA translocation activities. Surprisingly, the mutants can catalyze at least one ATP hydrolysis event but are unable to turn over and reset the motor. This is the first report of a catalytic block in translocating ATPase motor after ATP hydrolysis occurred. We suggest that the C-motif is an ATP hydrolysis sensor, linking product release to mechanical motion. A novel terminase-driven mechanism is proposed for translocation of dsDNA in viruses. PMID- 17428498 TI - Crystal structure of tryptophanyl-tRNA synthetase complexed with adenosine-5' tetraphosphate: evidence for distributed use of catalytic binding energy in amino acid activation by class I aminoacyl-tRNA synthetases. AB - Tryptophanyl-tRNA synthetase (TrpRS) is a functionally dimeric ligase, which specifically couples hydrolysis of ATP to AMP and pyrophosphate to the formation of an ester bond between tryptophan and the cognate tRNA. TrpRS from Bacillus stearothermophilus binds the ATP analogue, adenosine-5' tetraphosphate (AQP) competitively with ATP during pyrophosphate exchange. Estimates of binding affinity from this competitive inhibition and from isothermal titration calorimetry show that AQP binds 200 times more tightly than ATP both under conditions of induced-fit, where binding is coupled to an unfavorable conformational change, and under exchange conditions, where there is no conformational change. These binding data provide an indirect experimental measurement of +3.0 kcal/mol for the conformational free energy change associated with induced-fit assembly of the active site. Thermodynamic parameters derived from the calorimetry reveal very modest enthalpic changes, consistent with binding driven largely by a favorable entropy change. The 2.5 A structure of the TrpRS:AQP complex, determined de novo by X-ray crystallography, resembles that of the previously described, pre-transition state TrpRS:ATP complexes. The anticodon binding domain untwists relative to the Rossmann-fold domain by 20% of the way toward the orientation observed for the Products complex. An unexpected tetraphosphate conformation allows the gamma and deltad phosphate groups to occupy positions equivalent to those occupied by the beta and gamma phosphates of ATP. The beta-phosphate effects a 1.11 A extension that relocates the alpha phosphate toward the tryptophan carboxylate while the PPi mimic moves deeper into the KMSKS loop. This configuration improves interactions between enzyme and nucleotide significantly and uniformly in the adenosine and PPi binding subsites. A new hydrogen bond forms between S194 from the class I KMSKS signature sequence and the PPi mimic. These complementary thermodynamic and structural data are all consistent with the conclusion that the tetraphosphate mimics a transition-state in which the KMSKS loop develops increasingly tight bonds to the PPi leaving group, weakening linkage to the Palpha as it is relocated by an energetically favorable domain movement. Consistent with extensive mutational data on Tyrosyl tRNA synthetase, this aspect of the mechanism develops high transition-state affinity for the adenosine and pyrophosphate moieties, which move significantly, relative to one another, during the catalytic step. PMID- 17428499 TI - Neurospora crassa FKBP22 is a novel ER chaperone and functionally cooperates with BiP. AB - FK506 binding proteins (FKBPs) belong to the family of peptidyl prolyl cis-trans isomerases (PPIases) catalyzing the cis/trans isomerisation of Xaa-Pro bonds in oligopeptides and proteins. FKBPs are involved in folding, assembly and trafficking of proteins. However, only limited knowledge is available about the roles of FKBPs in the endoplasmic reticulum (ER) and their interaction with other proteins. Here we show the ER located Neurospora crassa FKBP22 to be a dimeric protein with PPIase and a novel chaperone activity. While the homodimerization of FKBP22 is mediated by its carboxy-terminal domain, the amino-terminal domain is a functional FKBP domain. The chaperone activity is mediated by the FKBP domain but is exhibited only by the full-length protein. We further demonstrate a direct interaction between FKBP22 and BiP, the major Hsp70 chaperone in the ER. The binding to BiP is mediated by the FKBP domain of FKBP22. Interestingly BiP enhances the chaperone activity of FKBP22. Both proteins form a stable complex with an unfolded substrate protein and thereby prevent its aggregation. These results suggest that BiP and FKBP22 form a folding helper complex with a high chaperoning capacity in the ER of Neurospora crassa. PMID- 17428500 TI - Thermodynamic and structural basis for transition-state stabilization in antibody catalyzed hydrolysis. AB - Catalytic antibodies 6D9 and 9C10, which were induced by immunization with a haptenic transition-state analog (TSA), catalyze the hydrolysis of a nonbioactive chloramphenicol monoester derivative to generate a bioactive chloramphenicol. These antibodies stabilize the transition state to catalyze the hydrolysis reaction, strictly according to the theoretical relationship: for 6D9, k(cat)/k(uncat)=895 and K(S)/K(TSA)=900, and for 9C10, k(cat)/k(uncat)=56 and K(S)/K(TSA)=60. To elucidate the molecular basis of the antibody-catalyzed reaction, the crystal structure of 6D9 was determined, and the binding thermodynamics of 6D9 and 9C10 with both the substrate and the TSA were analyzed using isothermal titration calorimetry. The crystal structure of the unliganded 6D9 Fab was determined at 2.25 A resolution and compared with that of the TSA liganded 6D9 Fab reported previously, showing that the TSA is bound into the hydrophobic pocket of the antigen-combining site in an "induced fit" manner, especially at the L1 and H3 CDR loops. Thermodynamic analyses showed that 6D9 binds the substrate of the TSA with a positive DeltaS, differing from general thermodynamic characteristics of antigen-antibody interactions. This positive DeltaS could be due to the hydrophobic interactions between 6D9 and the substrate or the TSA mediated by Trp H100i. The difference in DeltaG between substrate and TSA-binding to 6D9 was larger than that to 9C10, which is in good correlation with the larger k(cat) value of 6D9. Interestingly, the DeltaDeltaG was mainly because of the DeltaDeltaH. The correlation between k(cat) and DeltaDeltaH is suggestive of "enthalpic strain" leading to destabilization of antibody-substrate complexes. Together with X-ray structural analyses, the thermodynamic analyses suggest that upon binding the substrate, the antibody alters the conformation of the ester moiety in the substrate from the planar Z form to a thermodynamically unstable twisted conformation, followed by conversion into the transition state. Enthalpic strain also contributes to the transition-state stabilization by destabilizing the ground state, and its degree is much larger for the more efficient catalytic antibody, 6D9. PMID- 17428501 TI - Modeling the mechanisms of acute hepatitis B virus infection. AB - Mathematical models have been used to understand the factors that govern infectious disease progression in viral infections. Here we focus on hepatitis B virus (HBV) dynamics during the acute stages of the infection and analyze the immune mechanisms responsible for viral clearance. We start by presenting the basic model used to interpret HBV therapy studies conducted in chronically infected patients. We then introduce additional models to study acute infection where immune responses presumably play an important role in determining whether the infection will be cleared or become chronic. We add complexity incrementally and explain each step of the modeling process. Finally, we validate the model against experimental data to determine how well it represents the biological system and, consequently, how useful are its predictions. In particular, we find that a cell-mediated immune response plays an important role in controlling the virus after the peak in viral load. PMID- 17428502 TI - C(i,j) matrix: a better numerical characterization for graphical representations of biological sequences. AB - We find that the traditional numerical characterizations of biological sequences, such as E matrix, D/D matrix, L/L matrix and their "high order" matrices, have their limitations to characterize the biological sequences exactly, but they are widely used to analyze the biological sequences. Here, we propose a better numerical characterization for graphical representations of biological sequences, C(i,j) matrix. It is associated with the curvature of every point and has many advantages: (1) It can characterize the graphical representations for DNA sequences exactly, because it can overcome the limitation of the traditional matrices. (2) If we choose an appropriate fixed point, we can make the elements of the C(i,j) matrix less than or equal to 1. PMID- 17428503 TI - Optimal mix of screening and contact tracing for endemic diseases. AB - Two common means of controlling infectious diseases are screening and contact tracing. Which should be used, and when? We consider the problem of determining the cheapest mix of screening and contact tracing necessary to achieve a desired endemic prevalence of a disease or to identify a specified number of cases. We perform a partial equilibrium analysis of small-scale interventions, assuming that prevalence is unaffected by the intervention; we develop a full equilibrium analysis where we compare the long-term cost of various combinations of screening and contact tracing needed to achieve a given equilibrium prevalence; and we solve the problem of minimizing the total costs of identifying and treating disease cases plus the cost of untreated disease cases. Our analysis provides several insights. First, contact tracing is only cost effective when prevalence is below a threshold value. This threshold depends on the relative cost per case found by screening versus contact tracing. Second, for a given contact tracing policy, the screening rate needed to achieve a given prevalence or identify a specified number of cases is a decreasing function of disease prevalence. As prevalence increases above the threshold (and contact tracing is discontinued), the screening rate jumps discontinuously to a higher level. Third, these qualitative results hold when we consider unchanged or changed prevalence, and short-term or long-term costs. PMID- 17428504 TI - Modulation of the genotoxicity of bleomycin by amines through noncovalent DNA interactions and alteration of physiological conditions in yeast. AB - The effects of amines on the induction of mitotic gene conversion by bleomycin (BLM) were studied at the trp5 locus in Saccharomyces cerevisiae strain D7. BLM induces double-strand breaks in DNA and is a potent recombinagen in this assay. The polyamine spermidine causes concentration-dependent protection against the genotoxicity of BLM, reducing the convertant frequency by over 90% under the most protective conditions. Spermine, diethylenetriamine, ethylenediamine, putrescine, and ethylamine were also antigenotoxic in combined treatments with BLM. There was a general correspondence between the protective effect and the number of amino groups, suggesting that more strongly cationic amines tend to be stronger antirecombinagens. Electrostatic association of the amines with DNA probably hinders BLM access to the 4' position of deoxyribose where it generates a free radical. Other amines interact with BLM differently from these unbranched aliphatic amines. The aminothiol cysteamine inhibits the genotoxicity of BLM under hypoxic conditions but increases it under euoxic conditions. In contrast, pargyline potentiates the genotoxicity of BLM under hypoxic conditions but not under euoxic conditions. The antirecombinagenic effect of cysteamine apparently involves DNA binding and depletion of oxygen needed for BLM activity, whereas its potentiation of BLM entails its serving as an electron source for the activation of BLM. Pargyline may enhance BLM indirectly by preventing the depletion of oxygen by monoamine and polyamine oxidase. The planar 9-aminoacridine weakly induces gene conversion in strain D7, but it is strongly synergistic with BLM. Enhancement of BLM activity by this compound and by the related nitroacridine Entozon is apparently mediated by intercalation of the acridine ring system into DNA. Thus, the influence of amines on the genotoxicity of BLM in yeast encompasses antigenotoxic, potentiating, and synergistic interactions. The underlying mechanisms involve noncovalent association with DNA, altered BLM access to DNA, and modulation of physiological conditions. PMID- 17428505 TI - Potassium bromate treatment predominantly causes large deletions, but not GC>TA transversion in human cells. AB - Potassium bromate (KBrO(3)) is strongly carcinogenic in rodents and mutagenic in bacteria and mammalian cells in vitro. The proposed genotoxic mechanism for KBrO(3) is oxidative DNA damage. KBrO(3) can generate high yields of 8 hydroxydeoxyguanosine (8OHdG) DNA adducts, which cause GC>TA transversions in cell-free systems. In this study, we investigated the in vitro genotoxicity of KBrO(3) in human lymphoblastoid TK6 cells using the comet (COM) assay, the micronucleus (MN) test, and the thymidine kinase (TK) gene mutation assay. After a 4h treatment, the alkaline and neutral COM assay demonstrated that KBrO(3) directly yielded DNA damages including DNA double strand breaks (DSBs). KBrO(3) also induced MN and TK mutations concentration-dependently. At the highest concentration (5mM), KBrO(3) induced MN and TK mutation frequencies that were over 30 times the background level. Molecular analysis revealed that 90% of the induced mutations were large deletions that involved loss of heterozygosity (LOH) at the TK locus. Ionizing-irradiation exhibited similar mutational spectrum in our system. These results indicate that the major genotoxicity of KBrO(3) may be due to DSBs that lead to large deletions rather than to 8OHdG adducts that lead to GC>TA transversions, as is commonly believed. To better understand the genotoxic mechanism of KBrO(3), we analyzed gene expression profiles of TK6 cells using Affymetrix Genechip. Some genes involved in stress, apoptosis, and DNA repair were up-regulated by the treatment of KBrO(3). However, we could not observe the similarity of gene expression profile in the treatment of KBrO(3) to ionizing-irradiation as well as oxidative damage inducers. PMID- 17428507 TI - Mechanisms of regulation for interleukin-1beta in neurodegenerative disease. AB - The interleukin-1 family of cytokines are central to the pathology of acute and chronic diseases of the central nervous system. We describe current evidence on the transcriptional and post-transcriptional regulation of interleukin-1beta production, secretion and activity in the brain. Regarding the induction of protein synthesis, the possible involvement of Toll like receptor-4 is discussed including evidence that ischemic brain damage is reduced in Toll like receptor-4 knockout mice. The post-translational involvement of the P2X7-receptor and caspase-1 in the processing and release of active IL-1beta is also considered, as is evidence suggesting a possible extracellular cleavage of pro-IL-1beta by neutrophil derived proteases. We provide some fresh perspectives on how interleukin-1beta may be regulated and how these mechanisms could be targeted in disease. PMID- 17428506 TI - The role of tobacco smoke induced mitochondrial damage in vascular dysfunction and atherosclerosis. AB - The majority of individuals chronically exposed to tobacco smoke will eventually succumb to cardiovascular disease (CVD). However, despite the major cardiovascular health implications of tobacco smoke exposure, concepts of how such exposure specifically results in cardiovascular cell dysfunction that leads to CVD development are still being explored. Moreover, surprisingly little is known about the effects of prenatal and childhood tobacco smoke exposure on adult CVD development. Herein, it is proposed that the mitochondrion is a central target for environmental oxidants, including tobacco smoke. By virtue of its multiple, essential roles in cell function including energy production, oxidant signaling, apoptosis, immune response, and thermogenesis, damage to the mitochondrion will likely play an important role in the development of multiple common forms of human disease, including CVD. Specifically, this review will discuss the potential role of tobacco smoke and environmental oxidant exposure in the induction of mitochondrial damage which is related to CVD development. Furthermore, mechanisms of how mitochondrial damage can initiate and/or contribute to CVD are discussed, as are experimental results that are consistent with the hypothesis that mitochondrial damage and dysfunction will increase CVD susceptibility. Aspects of both adult and developmental (fetal and childhood) exposure to tobacco smoke on mitochondrial damage, function and disease development are also discussed, including the future implications and direction of studies involving the role of the mitochondrion in influencing disease susceptibility mediated by environmental factors. PMID- 17428508 TI - White matter volume predicts reaction time instability. AB - Information processing speed is a central concept in cognitive psychology and neuropsychology. Previous studies have mostly focused on mean reaction time (RT), and largely ignored intra-individual differences (the standard deviation of the RT: sdRT). Still, intra-individual inconsistency across trials has been shown to correlate with age, neurological disorders, intelligence, and performance on cognitive tests. However, sdRT has not been correlated with neuroanatomical variables. Such knowledge is important to the understanding of the neurobiological foundation for intra-individual variability. In the present study, white matter (WM) and cortical gray matter (GM) volume obtained from the average of two MR scans of 71 healthy participants (aged 20-88 years) were correlated with sdRT and mean RT obtained from a 3-stimulus visual oddball task. Negative correlations were hypothesized between sdRT and WM and between mean RT and cortical GM volume. These hypotheses were confirmed. The correlation between sdRT and WM volume was significant also independently of effects of age, gender, and mean RT, while there was a trend towards a significant correlation (p=.085) between cortical GM volume and mean RT independently of age. A path model was constructed, showing that age and sdRT gave independent contributions to the variance in performance intelligence, and that WM volume predicted performance score through the influence of sdRT. Further, sdRT was a stronger predictor of performance intelligence than mean RT. It is concluded that sdRT and mean RT may have different neuroanatomical correlates, and that sdRT is related to WM characteristics of the brain. PMID- 17428509 TI - Confabulation: motivated reality monitoring. AB - The study addressed the hypothesis that the content of confabulation is emotionally biased. Confabulating amnesic patients were compared with amnesic non confabulating patients in a memory recognition experiment that manipulated the valence (pleasant, unpleasant), temporal source (past, present, future) and selection agent (self, other) of the to-be-recognised memories. The results revealed that confabulating patients were more likely than amnesic non confabulating patients to incorrectly recognise past autobiographical events or thoughts as currently relevant memories, and this was more pronounced for pleasant compared to unpleasant events. These findings suggest that motivational factors, along with defective reality and temporality monitoring, contribute to confabulation. PMID- 17428510 TI - Assessment of positive emotions in animals to improve their welfare. AB - It is now widely accepted that good welfare is not simply the absence of negative experiences, but rather is primarily the presence of positive experiences such as pleasure. However scientific investigation of positive emotions has long been neglected. This paper addresses two main issues: first, it reviews the current state of scientific knowledge that supports the existence of positive affective states in animals and, second, it suggests possible applications of this knowledge that may enhance quality of life under animal management conditions. In the first part of the paper, recent advances in psychology and neuroscience are reviewed to provide pragmatic frameworks based on cognitive processes (such as positive anticipation, contrast and controllability) for further investigations of positive emotions in animals. Thereafter, the neurobiological bases of positive emotions are highlighted in order to identify behavioral and physiological expressions of positive experiences in animals. Monitoring both the autonomic nervous system (via heart rate and its variability) and the immune system could offer relevant tools to better assess emotional states in animals, complementary to classical adrenocortical measures. In the second part of the paper, useful strategies for enhancing positive experiences (such as physical, social and cognitive enrichment or putative genetic selection) are outlined. Then this paper emphasizes practical applications for assessing and promoting positive emotions that may help in providing animals with a better quality of life. Play, affiliative behaviors and some vocalizations appear to be the most promising convenient indicators for assessing positive experiences in laboratory and farm animals under commercial conditions. PMID- 17428511 TI - The acute effect of amylin and salmon calcitonin on energy expenditure. AB - The pancreatic B-cell hormone amylin is known to be involved in the regulation of meal ending satiation and it also shares typical features of adiposity signals. Chronic amylin administration has recently been shown to increase energy expenditure under certain conditions. Here we investigate the acute effect of peripheral administration of amylin or its agonist salmon calcitonin (sCT) on energy expenditure and respiratory quotient (RQ). First, rats were injected with amylin (5 microg/kg IP) or saline just before dark onset. Despite significantly decreased food intake in amylin-treated rats compared to control until 2 h post injection (p<0.05), amylin did not influence energy expenditure or RQ. Reduced food intake, which reduces energy expenditure, may have confounded a stimulatory effect of amylin on energy expenditure. Therefore, in the second experiment, amylin (1, 5 and 10 microg/kg IP) or saline was injected in the middle of the light phase (t=0 h) without access to food during 3 h post-injection. Amylin had no significant effects on energy expenditure or RQ. In a similar paradigm, the effect of sCT (0.1, 1.0 and 5.0 microg/kg IP) was tested. During food restriction, 5.0 microg/kg sCT significantly stimulated energy expenditure compared to control (p<0.05). Subsequent to refeeding at t=3 h, energy expenditure was decreased compared to control at t=8 h and t=10 h after 5.0 microg/kg sCT, probably due to sCT's strong anorectic action. Thus amylin may prevent the compensatory decrease in energy expenditure normally seen in animals that eat less. The longer acting sCT stimulated energy expenditure in animals without food access. PMID- 17428512 TI - Localisation and expression of a myelin associated neurite inhibitor, Nogo-A and its receptor Nogo-receptor by mammalian CNS cells. AB - Axon regeneration failure in the adult mammalian central nervous system (CNS) is partly due to inhibitory molecules associated with myelin. The Nogo receptor (NgR) plays a role in this process through an extraordinary degree of cross reactivity with three structurally unrelated myelin-associated inhibitory ligands namely; Nogo-A, myelin associated glycoprotein (MAG) and oligodendrocyte myelin glycoprotein (OMgp). The major aim of the study was to investigate and explore the cellular localisation and expression pattern of NgR and Nogo-A in the mammalian nervous system. We therefore generated a rabbit polyclonal anti-NgR antibody from the leucine rich repeat (LRR) No. 9 domain of the NgR polypeptide chain. Together with a commercially available polyclonal antibody specific for NgR, and in conjunction with double labeling immunofluorescence methods on cryosections and cell cultures, NgR immunoreactivity was observed in the CNS and dorsal root ganglia (DRG). In cellular populations, it was confined to neuronal cell bodies and their processes. NgR was also localised on the surface of extending DRG intact axons and growth cones in live staining experiments. Nogo-A, a member of the reticulon family protein, was widely distributed in the mammalian brain, spinal cord, and DRG. Intense Nogo-A immunoreactivity was also detected in oligodendrocyte cell bodies and their myelin sheaths in nerve fibre tracts of the CNS. Furthermore, numerous populations of neurons in the brain and spinal cord expressed Nogo-A to a variable extent in their cell bodies and neurites, suggesting additional, as-yet-unknown, functions of this protein. These results confirm results obtained by other researchers with different sets of antibodies. However, they also raise the question of the mechanism and circumstances under which NgR interacts with Nogo-A, as the latter appears to be confined to the cytoplasm and can therefore not be expected to bind NgR on the axon surface. PMID- 17428513 TI - Cardiac complications of a dengue fever outbreak in Sri Lanka, 2005. AB - A high incidence of cardiac complications was observed in an outbreak of dengue fever at General Hospital, Peradeniya, Sri Lanka, in 2005. This report describes 120 serologically confirmed dengue fever patients who presented during the outbreak. Seventy-five (62.5%) of these patients had electrocardiogram changes (T inversion, ST depression, bundle branch blocks) and were assigned to the 'cardiac group' (50 females, 25 males; median age 34 years, range 13-76). These patients were more susceptible to fatigue, dyspnoea, low peripheral oxygen saturation in room air (P=0.001), chest pain (P=0.001) and flushing of skin (P=0.05) than 45 (37.5%) patients who had normal electrocardiograms and made up the 'non-cardiac group'. In the cardiac group there were 31 primary and 44 secondary dengue patients. In the cardiac group, 17 (23%) patients had hypotension and 58 (77%) developed tachycardia and bradycardia (P<0.001) compared to four (9%) in the non cardiac group, suggestive of significant cardiac dysfunction. There was no correlation between pulse rate and body temperature: cardiac group (r=0.05; P=0.63); non-cardiac group (r=0.11, P=0.46). RT-PCR detected DEN-3 in three cardiac patients. PMID- 17428514 TI - Pfcrt haplotypes and in-vivo chloroquine response in Sundergarh district, Orissa, India. AB - The Plasmodium falciparum chloroquine resistance transporter (Pfcrt) K76T mutation and haplotype (amino acids 72-76) were analyzed as markers of chloroquine (CQ) resistance in the blood samples of patients from two sites of different intensities of malaria transmission (high, n=70; low, n=68) in Sundergarh district of Orissa, India and correlated with the in-vivo response. Early treatment failure (ETF) was significantly more frequent in the high endemic area (32.9 vs. 7.4%, P<0.001), with children below 5 years suffering more. A high frequency of pfcrt K76T mutation was observed in both the areas (87.1 vs. 79.4%, P=0.22). Patients carrying pfcrt 76T were the most likely to develop ETF (odds ratio 36; 95% CI 3.35-1653.3; P<0.001). The ratio of 76T:K76 was 22:9 and 11:14, respectively, in high and low endemic areas (odds ratio 3.1; 95% CI 0.9-11.03; P=0.04), which may be used as a measure of drug pressure. Sequences of pfcrt codons 72-76 showed 16 of the CQ-resistant haplotypes to be SVMNT, 5 CVMNT and 12 CVIET. The CQ-sensitive haplotypes were mostly CVMNK in 10 samples; CVIEK in 2 samples. Both Southeast Asian and South American haplotypes were present, with the latter predominating. PMID- 17428515 TI - Comparative study on the cell toxicity and enzymatic activity of two northern scyphozoan species Cyanea capillata (L.) and Cyanea lamarckii (Peron & Leslieur). AB - Two species of venomous pelagic cnidaria are compared according to their enzymatic, cytotoxic and haemolytic potency. The widely distributed jellyfish Cyanea capillata and Cyanea lamarckii were collected in the North Sea at the coasts of the Orkney Island and the Island of Helgoland. Purified cnidocyst extracts from fishing and mesenteric tentacles were prepared and tested for their bioactivity. The haemolysis induced by toxins of C. capillata was determined with respect to organism size and toxigenic organs. The haemolytic activity of the related species C. lamarckii was documented for the first time. Dose dependent haemolytic activities have been detected by means of protein equivalents at concentrations above 20mug(protein)/mL. Extracts of fishing tentacle cnidocysts showed a less potent haemolytic activity compared to extracts of mesenteric tentacles. In vitro studies with permanent cells of a hepatoma cell line have shown a time and concentration dependent loss of cell vitality up to 90% at 33.3mug(protein)/mL (10mug(protein)/10(5) cells). Supplementing the cell based toxicity tests an enzyme assay was performed to measure a phospholipase A(2) (PLA(2)) activity. A PLA(2)-like activity could be demonstrated in cnidocysts extracts prepared from mesenteric and fishing tentacles of both jellyfish species. PMID- 17428516 TI - Immunogenicity of hybrid DNA vaccines expressing hepatitis B core particles carrying human and simian immunodeficiency virus epitopes in mice and rhesus macaques. AB - An effective HIV vaccine will likely need to induce broad and potent CTL responses. Epitope-based vaccines offer significant potential for inducing multi specific CTL, but often require conjugation to T helper epitopes or carrier moieties to induce significant responses. We tested hybrid DNA vaccines encoding one or more HIV or SIV CTL epitopes fused to a hepatitis B core antigen (HBcAg) carrier gene as a means to improve the immunogenicity of epitope-based DNA vaccines. Immunization of mice with a HBcAg-HIV epitope DNA vaccine induced CD8(+) T cell responses that significantly exceeded levels induced with DNA encoding either the whole HIV antigen or the epitope alone. In rhesus macaques, a multi-epitope hybrid HBcAg-SIV DNA vaccine induced CTL responses to 13 different epitopes, including 3 epitopes that were previously not detected in SIV-infected macaques. These data demonstrate that immunization with hybrid HBcAg-epitope DNA vaccines is an effective strategy to increase the magnitude and breadth of HIV specific CTL responses. PMID- 17428517 TI - Entry inhibitor-based microbicides are active in vitro against HIV-1 isolates from multiple genetic subtypes. AB - Inhibitors of viral entry are under consideration as topical microbicides to prevent HIV-1 sexual transmission. Small molecules targeting HIV-1 gp120 (BMS 378806) or CCR5 (CMPD167), and a peptide fusion inhibitor (C52L), each blocks vaginal infection of macaques by a SHIV. A microbicide, however, must be active against multiple HIV-1 variants. We therefore tested BMS-C (a BMS-378806 derivative), CMPD167, C52L and the CXCR4 ligand AMD3465, alone and in combination, against 25 primary R5, 12 X4 and 7 R5X4 isolates from subtypes A-G. At high concentrations (0.1-1 microM), the replication of most R5 isolates in human donor lymphocytes was inhibited by >90%. At lower concentrations, double and triple combinations were more effective than individual inhibitors. Similar results were obtained with X4 viruses when AMD3465 was substituted for CMPD167. The R5X4 viruses were inhibited by combining AMD3465 with CMPD167, or by the coreceptor-independent compounds. Thus, combining entry inhibitors may improve microbicide effectiveness. PMID- 17428519 TI - Comparisons of water quality parameters from diverse catchments during dry periods and following rain events. AB - In this study, 12 catchments sites located along the north coast of New South Wales in Australia were grouped into the four categories of septic, cattle, sewage treatment plant (STP) and forested sites via cluster analysis based on their land use patterns. Water samples from all these sites were collected between October 2004 and June 2006 at a regular monthly interval and within 48 h of rain events. The samples were analyzed for bacterial counts including faecal coliform and total coliform; faecal sterols including coprostanol, epicoprostanol, cholesterol, cholestanol, 24-ethylcoprostanol, campesterol, stigmasterol and beta-sitosterol; and the elements including Na, Rb, Sr, Ag, Cd, Sn, Cs, Ba, Hg, Tl, Pb, Bi, U, Mg, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, K, As, Se, P and Mo. Over the course of the sampling period, the STP site had the highest average coprostanol level of 1693+/-567 ng/L which was significantly higher (p<0.05) than the septic sites (190+/-71 ng/L), the cattle sites (163+/-94 ng/L) and forested sites (14+/-4 ng/L). As expected, the forested sites had significantly lower average level of faecal coliforms (373+/-87 cfu/100 mL) compared with the STP (1395+/-574 cfu/100 mL), septic (1243+/-494 cfu/100 mL) and cattle sites (535+/-112 cfu/100 mL). The concentrations of coprostanol were not correlated with the numbers of faecal coliform bacteria when the entire data set was evaluated. The forested sites generally had the lowest average levels of elemental compositions, with significantly lower levels noted for Na, U, Mg, V, Cu, Sr, K, As, P and Mo, whereas Fe was the only element notably higher in the forested sites. Temporal and rain events analyses of the data set revealed that elevated levels of both coprostanol and faecal coliforms were not exclusive to rain events. The average coprostanol levels in rain event samples at each site were not significantly different compared with the corresponding dry event samples. Conversely, faecal coliform numbers increased by 2-4 times in rain events samples from septic, cattle and forested sites, but did not alter in the STP site. Multivariate analyses identified coprostanol and Sr as major contributing factors for the discrimination of septic, cattle, STP and forested sites for both rain and dry events samples. It was clear that each land use type of catchment could be characterized by biochemical, bacteriological and elemental parameters. PMID- 17428518 TI - Cell surface expression of CCR5 and other host factors influence the inhibition of HIV-1 infection of human lymphocytes by CCR5 ligands. AB - Several CCR5 ligands, including small molecules and monoclonal antibodies (MAbs), are being developed as therapies for infection with strains of human immunodeficiency virus type 1 (HIV-1) that use CCR5 for entry (R5 viruses). The efficacy of such therapies could be influenced by inter-individual differences in host factors, such as CCR5 expression levels. To study this, we used peripheral blood mononuclear cells (PBMCs) from humans and rhesus macaques. The half-maximal inhibitory concentrations (IC(50)) of the small-molecule CCR5 ligands CMPD167, UK427,857 and SCH-D, and of the PRO 140 MAb, differ by >2 logs in a donor dependent manner. We studied this variation by using flow cytometry to measure CCR5 expression on PBMCs from six of the human donors: the IC(50) values of both SCH-D and PRO 140 correlated with CCR5 expression (R(2)=0.64 and 0.99, respectively). We also determined the efficacy of the CCR5 ligands against HIV-1 infection of HeLa-derived cell lines that express CD4 at the same level but vary 2-fold in CCR5 expression (JC.48 and JC.53 cells). The moderately greater CCR5 expression on the JC.53 than the JC.48 cells was associated with proportionately higher median IC(50) values for all four CCR5 ligands but not for a soluble CD4 based inhibitor or a non-nucleoside reverse transcriptase inhibitor. We conclude that differences in CCR5 expression on human PBMCs, which can be affected by CCL3L1 gene dose, may influence the antiviral potency of CCR5 ligands in vitro, but other host factors are also likely to be involved. These host factors may affect the clinical activity of CCR5 inhibitors, including their use as topical microbicides to prevent HIV-1 transmission. PMID- 17428520 TI - Determining estrogenic steroids in Taipei waters and removal in drinking water treatment using high-flow solid-phase extraction and liquid chromatography/tandem mass spectrometry. AB - River water and wastewater treatment plant (WWTP) effluents from metropolitan Taipei, Taiwan were tested for the presence of the pollutants estrone (E1), estriol (E3), 17beta-estradiol (E2), and 17alpha-ethinylestradiol (EE2) using a new methodology that involves high-flow solid-phase extraction and liquid chromatography/tandem mass spectrometry. The method was also used to investigate the removal of the analytes by conventional drinking water treatment processes. Without adjusting the pH, we extracted 1-L samples with PolarPlus C18 Speedisks under a flow rate exceeding 100 mL/min, in which six samples could be done simultaneously using an extraction station. The adsorbent was washed with 40% methanol/60% water and then eluted by 50% methanol/50% dichloromethane. The eluate was concentrated until almost dry and was reconstituted by 20 microL of methanol. Quantitation was done by LC-MS/MS-negative electrospray ionization in the selected reaction monitoring mode with isotope-dilution techniques. The mobile phase was 10 mM N-methylmorpholine aqueous solution/acetonitrile with gradient elution. Mean recoveries of spiked Milli-Q water were 65-79% and precisions were within 2-20% of the tested concentrations (5.0-200 ng/L). The method was validated with spiked upstream river water; precisions were most within 10% of the tested concentrations (10-100 ng/L) with most RSDs<10%. LODs of the environmental matrixes were 0.78-7.65 ng/L. A pre-filtration step before solid-phase extraction may significantly influence the measurement of E1 and EE2 concentrations; disk overloading by water matrix may also impact analyte recoveries along with ion suppression. In the Taipei water study, the four steroid estrogens were detected in river samples (ca. 15 ng/L for E2 and EE2 and 35-45 ng/L for E1 and E3). Average levels of 19-26 ng/L for E1, E2, and EE2 were detected in most wastewater effluents, while only a single effluent sample contained E3. The higher level in the river was likely caused by the discharge of untreated human and farming waste into the water. In the drinking water treatment simulations, coagulation removed 20-50% of the estrogens. An increased dose of aluminum sulfate did not improve the performance. Despite the reactive phenolic moiety in the analytes, the steroids were decreased only 20-44% of the initial concentrations in pre- or post-chlorination. Rapid filtration, with crushed anthracite playing a major role, took out more than 84% of the estrogens. Except for E3, the whole procedure successfully removed most of the estrogens even if the initial concentration reached levels as high as 500 ng/L. PMID- 17428521 TI - Determination of estrogens and estrogenic activity in wastewater effluent by chemical analysis and the bioluminescent yeast assay. AB - A scheme of bioassay-directed analysis has been developed which combines a yeast assay screening for estrogenic activity with a liquid chromatographic-mass spectrometric (LC-MS/MS) chemical analysis, chromatographic fractionation, solid phase extraction and freeze-drying. The test scheme was applied on effluent samples collected from a municipal sewage treatment plant. The aim was to determine the substances responsible for main portion of the estrogenic activity in the samples and to compare the efficiency of different procedures for isolation and concentration of estogenicity. LC-MS/MS analyses were used for the quantification of 17beta-estradiol, estrone, estriol and 17alpha ethinylestradiol, and the measured concentrations compared with the activities found in the yeast assay. Following conversion of the concentrations measured by LC-MS/MS to 17beta-estradiol equivalents it was concluded that freeze-drying, solid phase extraction and the chemical analysis gave comparable activities. Since estrone was the major estrogen in the effluent, this estrogen was also the major contributor to the estrogenic activity in the effluent. The estrogenic activity was equivalent to 4-7 ng/L of 17beta-estradiol. The yeast assay results from the tests of the chromatographic fractions showed that the major activity resides in the fraction where estrone, 17beta-estradiol and 17alpha ethinylestradiol eluted. The activity of this fraction was substantially higher than the activity of the original wastewater sample. The reason for this could in part be explained by an inhibition of activity occurring in the original water sample. PMID- 17428522 TI - A relevant historical perspective on the current toxicity of IP chemotherapy: implications for the future. PMID- 17428523 TI - Endometrial carcinoma first presenting as paraneoplastic cerebellar degeneration. AB - BACKGROUND: Paraneoplastic degeneration (PCD) is an immune-mediated disorder affecting the cerebellum, characterized by subacute onset of cerebellar dysfunction progressing to severe disability, and is associated with an underlying malignancy. Ovarian carcinoma and breast carcinoma are commonly implicated. CASE: We report a rare case PCD in an elderly woman, later diagnosed with endometrial carcinoma. CONCLUSION: PCD is an unusual and rare manifestation of gynecological malignancies, including endometrial carcinoma, and a high degree of suspicion is required in these cases in order to make the correct diagnosis. PMID- 17428524 TI - The brain metabolic enhancer methylene blue improves discrimination learning in rats. AB - Methylene blue (MB) is a metabolic enhancer that has been demonstrated to improve memory retention when given post-training in low doses in a variety of tasks in rats, including inhibitory avoidance, spatial memory (in both normal and metabolically-impaired subjects), object recognition, and habituation to a familiar environment. MB has been also shown to improve memory retention of extinction of fear conditioning in the rat. No experiments have been conducted to determine the effects of MB on more complex learning such as in discrimination tasks that require repeated days of training. This study examined the effects of daily MB on spatial discrimination memory in a baited holeboard maze. Following three days of discrimination training, subjects treated daily with post-training MB (1 mg/kg) reliably discriminated between rewarded (baited) and non-rewarded (unbaited) trials as indicated by a greater number of correct responses on rewarded trials than non-rewarded trials during the last three days of discrimination training. No such discrimination effects were observed in the saline-treated control group during the same training period. To determine whether the memory-enhancing effects of MB are associated with an increase in metabolic energy capacity in the brain, cytochrome c oxidation was measured in brains from rats treated with 1 mg/kg MB or saline for three days. The number of daily injections was chosen based on the behavioral data which revealed group differences three days after the beginning of MB treatment. Brain cytochrome oxidase activity in the MB-treated group was approximately 70% higher than in saline-treated rats. The findings suggest that repeated post-training MB may improve memory consolidation between days of learning by an induction in the enzyme cytochrome oxidase, leading to increased metabolic capacity in brain regions requiring more energy during discrimination learning. PMID- 17428525 TI - Who needs liquor stores when parents will do? The importance of social sources of alcohol among young urban teens. AB - OBJECTIVES: Examine sources of alcohol over time in a large, ethnically diverse adolescent population from a poor, urban environment. METHODS: Surveys were administered at four time points (6th-8th grades) assessing demographic characteristics, past year alcohol use and sources of alcohol to youth in Chicago, Illinois 2002-2005. Growth curve analysis was used to examine alcohol access trends among all alcohol using youth and consistent alcohol users. Interactions by race and gender were tested. RESULTS: Social sources of alcohol were the most prevalent source over time. Parents were the primary source of alcohol, but their prominence significantly decreased over time. Taking alcohol from home, and getting alcohol from other adults, individuals under age 21, and commercial sources significantly increased as sources of alcohol over time. Males were significantly more likely than females to get alcohol from commercial sources and friends' parents. CONCLUSIONS: Greater attention for reducing social access to alcohol, particularly among parents, is needed for alcohol prevention efforts prior to and during middle school. PMID- 17428526 TI - Inadequate and infrequent are not alike: ERPs to deviant prosodic patterns in spoken sentence comprehension. AB - The current study on German investigates Event-Related brain Potentials (ERPs) for the perception of sentences with intonations which are infrequent (i.e. vocatives) or inadequate in daily conversation. These ERPs are compared to the processing correlates for sentences in which the syntax-to-prosody relations are congruent and used frequently during communication. Results show that perceiving an adequate but infrequent prosodic structure does not result in the same brain responses as encountering an inadequate prosodic pattern. While an early negative going ERP followed by an N400 were observed for both the infrequent and the inadequate syntax-to-prosody association, only the inadequate intonation also elicits a P600. PMID- 17428527 TI - Evaluation of narrative abilities in patients suffering from Duchenne Muscular Dystrophy. AB - The present work investigated cognitive, linguistic and narrative abilities in a group of children suffering from Duchenne Muscular Dystrophy, an allelic X-linked recessive disorder caused by mutations in the gene encoding dystrophin. The patients showed mildly reduced IQ with lower Verbal than Performance Intelligence Quotient and were mildly affected in visual attention and short-term memory processing. At the linguistic assessment, neither receptive (word comprehension) nor expressive (naming tasks and fluency) lexical abilities were impaired. However, their narratives were qualitatively inferior with respect to those produced by a group of typically developing children. Their speech samples were characterized by the presence of fewer verbs and complete sentences. It is suggested that the reduced production of complete sentences is due to a selective problem in verb argument structure generation. Since the lack of dystrophin is assumed to produce effects on the maturation of the cerebellum, whose involvement has been recently suggested in verb and syntactic processing, these findings may lend indirect support to the hypothesis of a cerebellar-cortical circuit specialized in verb and sentence production. PMID- 17428528 TI - The explicit learning of new names for known objects is improved by dexamphetamine. AB - A randomised, double-blind, placebo-controlled, between subjects study design (N=37) was used to investigate the effects of dexamphetamine on explicit new name learning. Participants ingested 10mg of dexamphetamine or placebo daily over 5 consecutive mornings before learning new names for 50 familiar objects plus fillers. The dexamphetamine group recognised and recalled the new names more accurately than the placebo group over the 5 days and 1 month later. Word learning success was not associated with baseline neuropsychological performance, mood, cardiovascular arousal, or sustained attention. These results may have implications for the pharmacological treatment of acquired naming difficulties. PMID- 17428529 TI - How aging affects the recognition of emotional speech. AB - To successfully infer a speaker's emotional state, diverse sources of emotional information need to be decoded. The present study explored to what extent emotional speech recognition of 'basic' emotions (anger, disgust, fear, happiness, pleasant surprise, sadness) differs between different sex (male/female) and age (young/middle-aged) groups in a behavioural experiment. Participants were asked to identify the emotional prosody of a sentence as accurately as possible. As a secondary goal, the perceptual findings were examined in relation to acoustic properties of the sentences presented. Findings indicate that emotion recognition rates differ between the different categories tested and that these patterns varied significantly as a function of age, but not of sex. PMID- 17428530 TI - Adhesion between dental ceramic and bonding resin: quantitative evaluation by Vickers indenter methodology. AB - OBJECTIVES: The purpose of this study was to evaluate the adhesion to dental ceramic by Vickers indenter methodology. This technique allows the creation of adhesive fractures and determines the influence of the surface treatment on adhesive capacities. METHODS: A single bond adhesive system (One Step Bisco) was applied to ceramic Vitapan 3D Master CE 0124 samples. Ceramic samples were polished with 500 or 4000-grit paper, sandblasted or not (Sa/NSa), treated with fluorhydric acid or not (A/NA) and silane or not (Si/NSi). The experimental groups (Gr) were: (Gr 1) 4000; (Gr 2) 4000+Si; (Gr 3) 4000+Sa+A; (Gr 4) 4000+Sa+A+Si; (Gr 5) 500+Sa+A+Si. Each sample was indented with the diamond Vickers indenter Leitz Durimet 2 (Wetzlar, Germany) using a load of 20N for 30s. The surfaces of the debonded areas were observed in an optical microscope providing a digital image of the debonded surface. The adhesion bond strength was calculated according to the formula of Engel and Roshon [Engel PA, Roshon DD. Indentation-debonding of an adhered surface layer. J Adhesion 1979;10(33): 237 53]. The statistical analysis was conducted using Student's t test (p<0.05). RESULTS: The values obtained for each group were: (Gr 1) 32MPa; (Gr 2) 52MPa; (Gr 3) 112MPa; (Gr 4) 131MPa; (Gr 5) 265MPa. There is a significant improvement in bond strength with the silane application on the 4000 polished surface (Gr 2). However, there is no significant difference when silane is applied or not on a sanded and etched ceramic (Gr 4). Bond strengths were higher with 500 grit polished, sanded, etched with silane application on the surface (Gr 5). SIGNIFICANCE: The Vickers indenter methodology is able to discriminate between the influences of different surface treatments on the adhesion of an adhesive layer on a feldspathic ceramic. PMID- 17428531 TI - Influence of molecular architecture of polyether-co-polyester dendrimers on the encapsulation and release of methotrexate. AB - In the present study, effects of alterations in the chemical structure of polyester-co-polyether (PEPE) dendrimers on the encapsulation and release of methotrexate (MTX) was investigated. A series of PEPE dendrimers of different architecture were synthesized. Biocompatibility of the resulting dendrimers was evaluated in vitro by assessing their cytotoxicity on RAW 264.7 cells using lactate dehydrogenase (LDH) assay. Dendrimers caused no cell death even at the concentration of 250 microg/mL, suggesting that they are acceptable for pharmaceutical applications. They also showed good capacity to encapsulate MTX, with loading as high as 24.5% w/w. Increase in the number of branches and the size of internal voids were shown to enhance the encapsulation. On the other hand, absence of aromatic rings as branching units drastically reduced the loading capacity. Physical entrapment, weak hydrogen bonding and hydrophobic interactions were established to be the mechanisms of encapsulation. Release of MTX was biphasic, which included a burst release in 6h followed by a slower release over a period of 50 or 168 h. Increase in the number of branches profoundly decreased this initial burst release; in contrast, absence of aromatic rings in the dendritic structure resulted in a very rapid release. PMID- 17428532 TI - Effect of wettability and surface functional groups on protein adsorption and cell adhesion using well-defined mixed self-assembled monolayers. AB - Self-assembled monolayers (SAMs) of alkanethiols, which can provide flat and chemically well-defined surfaces, were employed as model surfaces to understand cellular interaction with artificial materials. SAMs presenting a wide range of wettabilities were prepared by mixing two kinds of alkanethiols carrying terminal methyl (CH(3)), hydroxyl (OH), carboxylic acid (COOH), or amino (NH(2)) groups. Adhesion behavior of human umbilical vein endothelial cells (HUVECs) and HeLa cells on these mixed SAMs were examined. The number of adhered HUVECs reached a maximum on CH(3)/OH mixed SAMs with a water contact angle of 40 degrees , while cell adhesion increased with decreasing water contact angle up to 60-70 degrees and then leveled off on CH(3)/COOH and CH(3)/NH(2) mixed SAMs. Numbers of adhered HeLa cells showed a maximum on CH(3)/OH and CH(3)/COOH mixed SAMs with a water contact angle of 50 degrees . These facts suggest that cell adhesion is mainly determined by surface wettability, but is also affected by the surface functional group, its surface density, and the kinds of cells. The effect of exchange of adsorbed proteins on cell adhesion was also examined. HUVECs were cultured on the mixed SAMs preadsorbed with albumin. Cell adhesion was effectively prohibited on hydrophobic SAMs pretreated with albumin. Albumin strongly adsorbed and resisted replacement by cell adhesive proteins on hydrophobic SAMs. On the other hand, cells adhered to albumin-adsorbed hydrophilic SAMs. Displacement of preadsorbed albumin with cell adhesive proteins effectively occurs on these hydrophilic SAMs. This effect contributes to induce SAMs with moderate wettability to give suitable surfaces for cell adhesion. PMID- 17428533 TI - Si-C linked oligo(ethylene glycol) layers in silicon-based photonic crystals: optimization for implantable optical materials. AB - Porous silicon has shown potential for various applications in biology and medicine, which require that the material (1) remain stable for the length of the intended application and (2) resist non-specific adsorption of proteins. Here we explore the efficacy of short oligo(ethylene glycol) moieties incorporated into organic layers via two separate strategies in achieving these aims. In the first strategy the porous silicon structure was modified in a single step via hydrosilylation of alpha-oligo(ethylene glycol)-omega-alkenes containing three or six ethylene glycol units. The second strategy employs two steps: (1) hydrosilylation of succinimidyl-10-undecenoate and (2) coupling of an amino hexa(ethylene glycol) species. The porous silicon photonic crystals modified by the two-step strategy displayed greater stability relative to the single step procedure when exposed to conditions of physiological temperature and pH. Both strategies produced layers that resist non-specific adsorption of proteins as determined with fluorescently labelled bovine serum albumin. The antifouling behaviour and greater stability to physiological conditions provided by this chemistry enhances the suitability of porous silicon for biomaterials applications. PMID- 17428534 TI - Assembled alginate/chitosan nanotubes for biological application. AB - Biodegradable nanotubes were fabricated through the layer-by-layer (LbL) assembly technique of alternate adsorption of alginate (ALG) and chitosan (CHI) onto the inner pores of polycarbonate template with the subsequent removal of the template. The assembled materials present good film formation ability. The thickness of nanotubes wall can be controlled by changing the assembled layers. The assembled tubular structure was verified by the confocal laser scanning microscope (CLSM) using fluorescent-labeled ALG as well as the measurements of scan electron microscope (SEM) and transmission electron microscopy (TEM). Atomic force microscopy (AFM) images confirm the biodegradable feature of the assembled nanotubes as they are immersed in the pancreatin. Confocal microscopy images show that the assembled ALG/CHI nanotubes can be internalized into the cancer cell readily. The cell viability experiment proves the low cytotoxicity of ALG/CHI nanotubes. The final assembled nanotubes have presented good biodegradability and low cytotoxicity. PMID- 17428535 TI - A guide to sparkology: the taxonomy of elementary cellular Ca2+ signaling events. AB - Since the discovery of the Ca(2+) spark as an elementary event of cellular Ca(2+) signaling almost 15 years ago, the family of newly described Ca(2+) signal entities has been ever growing. While scientists working in Ca(2+) signaling may have maintained an overview over the specifics of this nomenclature, those outside the field often make the complaint that they feel hopelessly lost. With the present review we collect and summarize systematic information on the many Ca(2+) signaling events described in a variety of tissues and cells, and we emphasize why and how each of them has its own importance. Most of these signals are taking place in the cytosol of the respective cells, but several events have been recorded from intracellular organelles as well, where they may serve their own specific functions. Finally, we also try to convey an integrated view as to why cellular microdomain signaling is of fundamental biological importance. PMID- 17428536 TI - Brefeldin A triggers apoptosis associated with mitochondrial breach and enhances HA14-1- and anti-Fas-mediated cell killing in follicular lymphoma cells. AB - Follicular lymphoma (FL) remains a fatal disease of increasing worldwide incidence. Since patients with FL eventually develop resistance to conventional anticancer agents, and due to BCL-2 overexpression present with profoundly compromised execution of mitochondrial pathway of apoptosis, targeting alternative pathways of cell demise may appear therapeutically beneficial. Herein we report for the first time the effects of an ER-Golgi transport inhibitor, Brefeldin A (BFA), alone and in combination with a small molecule Bcl-2 inhibitor HA14-1 or agonistic anti-Fas mAb, in the recently established human FL cell lines. All cell lines tested were sensitive to BFA-induced cytotoxicity and apoptosis. Moreover BFA-induced cell death was associated with profound ER stress, mitochondrial breach and subsequent caspase cascade activation, including caspase 2 activation. Interestingly, BFA-induced ER stress did not result in appearance of autophagic morphology in FL cells. Of importance, small molecule Bcl-2 antagonist, HA14-1 and agonistic anti-Fas mAb significantly enhanced BFA mediated cytotoxicity and apoptosis, revealing novel and previously unexplored means to enhance ER stress-mediated cell killing in follicular lymphoma cells. PMID- 17428537 TI - Therapeutic efficacy of NSC606985, a novel camptothecin analog, in a mouse model of acute promyelocytic leukemia. AB - Nanomolar concentrations of NSC606985, a novel camptothecin (CPT) analog, effectively induces apoptosis in vitro in acute myeloid leukemic (AML) cells. Here we investigated the potential therapeutic effects of NSC606985 on the mice model with acute promyelocytic leukaemia (APL), a unique subtype of AML. The results showed that NSC606985 at single dose rapidly eliminated the infiltration with apoptosis induction of leukemic cells in peripheral blood and tissues. Treatment of NSC606985 with two regimens, continuous monotherapy and intermittent long-term therapy, significantly prolonged the survival of leukemic mice with tumor regression. These results propose that NSC606985 warrants further preclinical and clinical investigations for AML treatment. PMID- 17428538 TI - Ontogenesis of haemocytes in shrimp (Fenneropenaeus chinensis) studied with probes of monoclonal antibody. AB - Monoclonal antibodies (Mabs) specifically against haemocytes of shrimp (Fenneropenaeus chinensis) were employed to study the ontogenesis of haemocytes by immunofluorescence assay technique (IFAT) with the aids of avidin-biotin complex method. Different developmental stages were identified by means of histochemical staining. Stages including blastula, gastrula, limb bud stage, embryonic-nauplius and nauplius were examined. With Mab 2A3 against both hyline, semi-granular and granular haemocytes, the first positive cells were detected at the limb bud stage of the embryos. Distinguishable positive haemocytes with a diameter of about 7 microm appeared in embryonic-nauplius. Whereas, with Mab 1H11 specific to cytoplasmic granulars in haemocytes, the first positive granular haemocytes were observed at the stage of embryonic-nauplius. Increased number of both total and granular haemocytes at the stage of nauplius impressed a possible enhancement of the immune system as the embryogenesis progressed. PMID- 17428539 TI - Rickettsia peacockii, an endosymbiont of Dermacentor andersoni, does not elicit or inhibit humoral immune responses from immunocompetent D. andersoni or Ixodes scapularis cell lines. AB - Ixodes scapularis and Dermacentor andersoni cell lines were stimulated with heat killed Escherichia coli and Micrococcus luteus to investigate whether infection by Rickettsia peacockii, an endosymbiont of D. andersoni, modifies humoral immune responses. Radial diffusion assays, western blotting, flow cytometry, and quantitative reverse-transcription PCR were used to determine if expression of bacteriolytic peptides, including lysozyme and defensin, was upregulated by bacterial stimulation or infection with R. peacockii. The I. scapularis line IDE12 upregulated expression of lysozyme and defensin following stimulation. The D. andersoni cell line DAE15 also expressed defensin and lysozyme, but only lysozyme was upregulated by bacterial stimulation. R. peacockii infection alone, or in cells stimulated with bacteria, did not modify defensin or lysozyme expression in either cell line. These results suggest tick endosymbionts may avoid recognition by the tick immune system, and infection may not affect humoral immune responses to bacteria not normally associated with ticks. PMID- 17428540 TI - Roman concept of mental capacity to make end-of-life decisions. AB - When assessing decisional competence of patients, psychiatrists have to balance the patients' right to personal autonomy, their condition and wishes against principles of medical ethics and professional discretion. This article explores the age-old legal and ethical dilemmas posed by refusal of vital medical treatment by patients and their mental capacity to make end-of-life decisions against the background of philosophical, legal and medical approaches to these issues in the time of the Younger Pliny (c62-c113 CE). Classical Roman discourse regarding mental competency and "voluntary death" formed an important theme of the vast corpus of Greco-Roman writings, which was moulded not only by legal permissibility of suicide but also by philosophical (in modern terms, moral or ethical) considerations. Indeed, the legal and ethical issues of evaluating the acceptability of end of life decisions discussed in the Letters are as pertinent today as they were 2000 years ago. We may gain valuable insights about our own methodologies and frames of reference in this area of the law and psychiatry by examining Classical Roman approaches to evaluating acceptability of death-choices as described in Pliny's Letters and the writings of some of his peers. PMID- 17428541 TI - Pharmacokinetics and tissue distribution of novel traditional Chinese medicine platinum anticancer agents in rats. AB - The pharmacokinetics and tissue distribution profiles of a novel series of traditional Chinese medicine-platinum (TCM-Pt) compounds [Pt(C(8)H(8)O(5))(NH(2)R)(2)]: 1 (where R=H), 3 (R=CH(3)) and 5 (R=C(6)H(10)), were studied in Sprague-Dawley rats following a single bolus intravenous (i.v.) injection. Platinum concentrations in total plasma, plasma ultrafiltrate, urine and tissues were measured by flameless atomic absorption spectroscopy. Pharmacokinetic studies showed that plasma concentrations of total and free platinum for the novel TCM-Pt compounds as well as cisplatin and carboplatin declined in a biexponential manner with a short distribution half-life (t(1/2alpha): 0.12-0.34h). Compared with cisplatin, the novel TCM-Pt compounds had a longer elimination half-life (t(1/2beta)), larger dose normalized area under the curve (AUC/D), larger volume of distribution at steady-state (V(ss)), slower clearance (CL) of free platinum and higher percentage of cumulative urinary excretion (CUE), which can be attributed to their lower chemical reactivities. In tissues, the highest Pt concentrations were found in the kidney, followed by the liver and the lowest in the heart; no Pt was detected in the brain. Twenty-four hours after drug administration, platinum concentrations in tissues were significantly lower for the novel TCM-Pt compounds. These findings suggest that the novel compounds might afford higher clinical efficacy and reduced systemic side effects, when compared with cisplatin. PMID- 17428542 TI - QuantiFERON-TB Gold test in the identification of latent tuberculosis infection in immigrants. AB - BACKGROUND: The identification and treatment of latent tuberculosis infection (LTBI) among immigrants are an effective strategy for TB control in developed countries. A new test for LTBI identification that uses more specific antigens of Mycobacterium tuberculosis is now commercially available under the brand name of QuantiFERON-TB Gold test. OBJECTIVE: To compare QuantiFERON-TB Gold test to tuberculin skin testing (TST) for the detection of LTBI among immigrants from high endemic TB areas. PATIENTS AND METHODS: Undocumented immigrants attending a district medical center were enrolled if they originated from high endemic TB areas, the time of arrival in Italy was < or = 5 years, had neither active TB disease nor known immunodeficiency status. The TST was applied according to standards and QuantiFERON-TB Gold test was performed following the manufacturer's instructions. RESULTS: Hundred subjects were included in the comparative analysis. TST was positive in 44% of subjects; 15% had a positive QuantiFERON-TB Gold test result. The total agreement between TST and QuantiFERON-TB Gold test was 71%, for a kappa statistics of 0.37; agreement was 100% for TST negative results, but only 34% for TST positive ones. In the multivariate logistic regression analysis, previous BCG vaccination was independently associated with a lower chance of disagreement between the tests. CONCLUSION: The prevalence of LTBI among immigrants was lower when determined by QuantiFERON-TB Gold; this may be a consequence of more specific MTB antigens used. Our results suggest that QuantiFERON-TB Gold may be used as confirmatory test for TST positive immigrants candidate to preventive therapy. PMID- 17428543 TI - Parent-infant interaction: a growth model approach. AB - The aim of this study is to compare the interaction of fathers and mothers with their 10-12 months old infants (n=97; parental sensitivity and mood, and infant mood) during five structured contiguous play segments, and to examine the utility of individual growth modeling. Conventional comparison of means across play segments showed that parents were equally responsive, but mothers were happier than fathers, and infants were equally happy during interaction with both parents. Sensitivity and mood were more strongly related for mothers than for fathers. Uni- and multivariate growth models revealed fine-grained patterns not seen in conventional analysis: (a) parental and infant mood decreased across play more for mothers than for fathers, (b) parental sensitivity in one play-segment predicted parental mood and infant mood in the next segment, (c) change in infants' mood was related to change in sensitivity in mothers, and to change in mood in fathers, and (d) mothers' sensitive interaction with the infant was predicted by family socio-demographic background. PMID- 17428544 TI - Temperament and personality in bipolar I patients with and without mixed episodes. AB - BACKGROUND: Personality and temperament are supposed to have an impact on the clinical expression and course of an affective disorder. There is some indication, that mixed episodes result from an admixture of inverse temperamental factors to a manic syndrome. In a preliminary report [Brieger, P., Roettig, S., Ehrt, U., Wenzel, A., Bloink, R., Marneros, A., 2003. TEMPS-a scale in 'mixed' and 'pure' manic episodes: new data and methodological considerations on the relevance of joint anxious-depressive temperament traits. J. Affect. Disord. 73, 99-104] we reported support for this assumption. The present study completes the preliminary results and compares patients with and without mixed episodes with respect to personality and personality disorders in addition. METHODS: Patients who had been hospitalized for bipolar I disorder were reassessed after 4.8 years. We examined temperament (TEMPS-A), personality (NEO-FFI) and frequency of personality disorders (SCID-II). Furthermore, illness-related parameters like age at first treatment, depressive and manic symptomatology, frequency and type of episodes and level of functioning were obtained and patients with and without mixed episodes were compared. RESULTS: Patients with (n=49) and without mixed episodes (n=86) did not differ significantly with regard to the illness-related parameters and personality dimensions. The frequency of personality disorders was significantly higher in patients with prior mixed episodes. With respect to temperament, scores of the depressive, cyclothymic, irritable and anxious temperament were significantly higher in patients with mixed episodes. LIMITATIONS: We were not able to assess premorbid temperament and premorbid personality. CONCLUSIONS: The findings of the present study support the assumption of Akiskal [Akiskal, H.S., 1992b. The distinctive mixed states of bipolar I, II, and III. Clin. Neuropharmacol. 15 Suppl 1 Pt A, 632-633.] that mixed episodes are more frequent in subjects with inverse temperament. PMID- 17428545 TI - Polymorphisms of CD1 genes in chronic dysimmune neuropathies. AB - CD1 are MCH-like glycoproteins specialized in capturing and presenting glycolipid to T cells. Expression of CD1 molecules has been observed on endoneurial machrophages in patients with chronic inflammatory demyelinating polyneuropathy (CIDP) and vasculitis and polymorphisms of CID1A and CD1E genes have been associated with susceptibility to develop Guillain-Barre syndrome. In 46 patients with CIDP, in 13 patients with multifocal motor neuropathy and in 132 controls we genotyped exon 2 of CD1A and CD1E genes. We found no association between chronic dysimmune neuropathies, with or without anti-ganglioside antibodies, and polymorphisms of CD1A and CD1E genes. PMID- 17428546 TI - Enhanced cell-to-cell contacts between activated microglia and pyramidal cell dendrites following kainic acid-induced neurotoxicity in the hippocampus. AB - Microglia participate in immune responses in the brain. However, little is known about the contact-mediated interaction between microglia and neurons. We report here that the cell-to-cell contacts between microglial processes and dendrites of hippocampal CA1 neurons were dramatically increased in density and area following local injection of kainic acid (KA). A similar KA-induced increase in the degree of intercellular contacts was observed in mice lacking telencephalin (TLCN), a neuronal dendritic adhesion molecule of ICAM family. The results suggest that adhesive contacts independent of TLCN and contact-mediated interactions between microglia and dendrites were promoted by excitotoxic brain injury. PMID- 17428547 TI - Aggravation of experimental autoimmune neuritis in TNF-alpha receptor 1 deficient mice. AB - The role of tumor necrosis factor (TNF)-alpha and its receptors in the pathogenesis of experimental autoimmune neuritis (EAN) induced by P0 peptide 180 199 in TNFR1 (p55) deficient (TNFR1-/-) mice was investigated. Compared to wild type EAN mice, TNFR1-/- EAN mice developed significantly more severe clinical signs, in parallel with enhanced numbers of inflammatory infiltrating cells in peripheral nerves and splenic P0-reactive T cell proliferation, as well as increased obviously MHC class II and CCR3 expression on the macrophages in the cauda equina. Our data indicated that TNF-alpha might have anti-inflammatory effect preventing the development of EAN in this mouse model. PMID- 17428548 TI - Murine hematopoietic stem cells and progenitors express adrenergic receptors. AB - Association between the nervous and immune system is well documented. Immune cells originate within the bone marrow that is innervated. Thermal injury induces adrenergic stimulation, augments monocytopoiesis and alters the beta-adrenergic receptor (AR) profile of bone marrow monocyte committed progenitors. This provides an impetus to study AR expression in hematopoietic progenitors along myeloid lineage. Using FACS analysis and confocal microscopy, we report the expression of alpha1-, alpha2- and beta(2)-AR in enriched populations of ER MP209(+) and ER-MP12(+) myeloid progenitors, CD117(+) and CD34(+) multi-potential progenitors and more importantly pluripotent stem cells suggesting a plausible role for catecholamine in hematopoietic development. PMID- 17428549 TI - Interferon-alpha effects are exaggerated when administered on a psychosocial stressor backdrop: cytokine, corticosterone and brain monoamine variations. AB - Immunotherapy involving interferon-alpha (IFN-alpha) treatment is often accompanied by symptoms of depressive illness. These effects may stem from the direct actions of the cytokine, or may be unique to individuals undergoing considerable strain. In two experiments using CD-1 mice, we demonstrate that intraperitoneal administration of IFN-alpha dose dependently influences plasma corticosterone and sickness behaviors, and modestly influences norepinephrine turnover in brain. However, when mice are exposed to a psychosocial stressor (social disruption by transferring mice from isolated to grouped conditions, and to a moderate extent a transfer from grouped housing to isolation), the effects of IFN-alpha on sickness, plasma corticosterone and hippocampal norepinephrine, as well as on the levels of circulating IL-6, TNF-alpha and IL-10 (but not IL 1beta or IFN-gamma) are greatly augmented. It is suggested that the depressive effects of immunotherapy in humans likewise reflects the synergistic actions of the cytokine and the ongoing distress experienced by patients. PMID- 17428550 TI - High carrier frequency of the GJB2 mutation (35delG) in the north of Iran. AB - OBJECTIVE: Mutations in the GJB2 gene are a major cause of autosomal recessive and sporadic non-syndromic hearing loss in many populations. A single mutation of this gene (35delG) accounts for approximately 70% of mutations in Caucasians with a carrier frequency of 2-4% in Europe. This study aims to determine the rate of 35delG carrier frequency in Iran. METHODS: Genomic DNA was extracted from a total of 550 unaffected unrelated subjects from 4 provinces of Iran following the standard phenol chloroform procedure. The one base pair deletion (35delG) was analysed using a nested PCR procedure; 35delG mutation carriers were subsequently confirmed by sequence analysis. Moreover, using the Binomial probability distribution, we compared the 35delG carrier frequency of Iranian population with the various Middle Eastern and overall European populations. RESULTS: Of the four populations studied, we found a high carrier frequency of 2.8% in Gilan province in the north of Iran. The overall 35delG carrier frequency was found to be 1.25% in the populations studied (our present and previous data) which is similar to the overall 35delG carrier frequency detected in Middle Eastern populations, but Significantly lower than that identified in European populations. PMID- 17428551 TI - Severity of juvenile onset recurrent respiratory papillomatosis is not associated with socioeconomic status in a setting of universal health care. AB - BACKGROUND: Juvenile onset recurrent respiratory papillomatosis (JORRP) results from HPV transmission. Cervical cancer, also transmitted via HPV, is known to be correlated with socioeconomic status (SES). This study aims to determine if an association exists between SES and severity of JORRP. METHODS: Cross-sectional study of all active JORRP patients at the Hospital for Sick Children in Toronto in 2005. SES information from Hollingshead surveys, Postal walk Census data, and Low Income Cutoff Data were compared with Derkay-Wiatrak disease severity scores, peak annual surgical frequency, and age of diagnosis. Statistical analysis was performed using Spearman, Mann-Whitney, and linear regression analyses. RESULTS: Twenty-one patients were surveyed. Hollingshead results were as follows: two patients (10%) were class I (major business and professional); 11 patients (52%) were class II (medium business, minor professional, technical); 4 patients (19%) were class III (skilled craftsmen, clerical, sales workers); 4 patients (19%) were class IV (machine operators, semiskilled workers); 0% were from class V (unskilled laborers, menial service workers). Interestingly, based on postal code data nine patients (45%) were below the low income cutoff as compared to the Toronto (metropolitan) and Ontario (provincial) rates of low income (17% and 14%, respectively). There was significant correlation between each of the SES measures and between disease severity measures. However, analysis of the SES measures versus disease severity measures did not demonstrate any significant relationship. CONCLUSIONS: Though almost half the patients lived below the low income cutoff, this study did not demonstrate a significant correlation between socioeconomic status and severity of disease in JORRP. One possible explanation is that universal access to the Canadian health care system is able to provide support despite a large proportion of patients being socioeconomically vulnerable. A national level study is underway to further detect any relationship between SES and JORRP severity in the general population. PMID- 17428552 TI - Development of an oligonucleotide-based DNA microarray for transcriptional analysis of Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) genes. AB - A modified oligonucleotide-based two-channel DNA microarray was developed for characterization of temporal expression profiles of select Choristoneura fumiferana nucleopolyhedrovirus (CfMNPV) ORFs including its 7 unique ORFs. The microarray chip contained oligonucleotide probes for 23 CfMNPV ORFs and their complements as well as five host genes. Total RNA was isolated at different times post infection from Cf203 insect cells infected with CfMNPV. The cDNA was synthesized, fluorescent labelled with Cy3, and co-hybridized to the microarray chips along with Cy5-labelled viral genomic DNA, which served as equimolar reference standards for each probe. Transcription of the 7 CfMNPV unique ORFs was detected using DNA microarray analysis and their temporal expression profiles suggest that they are functional genes. The expression levels of three host genes varied throughout virus infection and therefore were unsuitable for normalization between microarrays. The DNA microarray results were compared to quantitative RT PCR (qRT-PCR). Transcription of the non-coding (antisense) strands of some of the CfMNPV select genes including the polyhedrin gene, was also detected by array analysis and confirmed by qRT-PCR. The polyhedrin antisense transcript, based on long-range RT-PCR analysis, appeared to be a read-through product of an adjacent ORF in the same orientation as the antisense transcript. PMID- 17428553 TI - Plant lectins are potent inhibitors of coronaviruses by interfering with two targets in the viral replication cycle. AB - We describe the antiviral activity of plant lectins with specificity for different glycan structures against the severe acute respiratory syndrome coronavirus (SARS-CoV) and the feline infectious peritonitis virus (FIPV) in vitro. The SARS-CoV emerged in 2002 as an important cause of severe lower respiratory tract infection in humans, and FIPV infection causes a chronic and often fatal peritonitis in cats. A unique collection of 33 plant lectins with different specificities were evaluated. The plant lectins possessed marked antiviral properties against both coronaviruses with EC(50) values in the lower microgram/ml range (middle nanomolar range), being non-toxic (CC(50)) at 50-100 microg/ml. The strongest anti-coronavirus activity was found predominantly among the mannose-binding lectins. In addition, a number of galactose-, N acetylgalactosamine-, glucose-, and N-acetylglucosamine-specific plant agglutinines exhibited anti-coronaviral activity. A significant correlation (with an r-value of 0.70) between the EC(50) values of the 10 mannose-specific plant lectins effective against the two coronaviruses was found. In contrast, little correlation was seen between the activity of other types of lectins. Two targets of possible antiviral intervention were identified in the replication cycle of SARS-CoV. The first target is located early in the replication cycle, most probably viral attachment, and the second target is located at the end of the infectious virus cycle. PMID- 17428554 TI - The influence of interleukin-1beta on oxytocin signalling in primary cells of human decidua. AB - OBJECTIVE: Oxytocin (OT) and its corresponding receptor (OTR), synthesized within the pregnant uterus, play a key role in the process of (preterm) labor as part of a paracrine system that regulates symmetrical contractility. In the setting of intrauterine infection, a major cause of preterm labour and birth, decidua serves as a major source of cytokine production. The present study evaluates the time dependent effect [0-24 h] of the inflammatory cytokine Interleukin-1beta (IL 1beta) treatment on OT signalling and OT stimulated prostaglandin release in primary cultures of human decidua. STUDY DESIGN: Primary cultures of human decidua (n=6) were treated with IL-1beta [5 ng/ml] for 0-24h and or indomethacin [100 microM]--an inhibitor of the prostaglandin synthesis--for 0-24 h or for 24 h. OT peptide expression, OTR binding, Inositol triphosphate production (IP(3)), and arachidonic acid (AA) as well as prostaglandin (PGE(2)) release were measured. RESULTS: IL-1beta transiently reduced cytoplasmic OT peptide at 4-6 h of IL-1beta incubation, while its secretion into the media was increased after 6 h of stimulation. The later was completely blocked by indomethacin. A decrease in OTR mRNA expression and a loss of OTR binding were detected after 8 h and 16 h of IL-1beta treatment, respectively. IL-1beta also decreased IP(3) production and AA release, but significantly enhanced OT mediated PGE(2) production. This effect was completely suppressed by the cyclooxygenase-2 (COX-2) inhibitor NS-398. CONCLUSION: Our data suggest, that IL-1beta indirectly increases OT secretion in primary cultures of human decidua in a time dependent fashion through the production of prostaglandins through COX-2 and that this increase in OT peptide may secondarily down-regulate the OTR and its signalling cascade. These findings might explain the poor effectiveness of oxytocin receptor antagonists as tocolytic agents in the setting of intrauterine infection. PMID- 17428555 TI - Ras oncogenes and their downstream targets. AB - RAS proteins are small GTPases, which serve as master regulators of a myriad of signaling cascades involved in highly diverse cellular processes. RAS oncogenes have been originally discovered as retroviral oncogenes, and ever since constitutively activating RAS mutations have been identified in human tumors, they are in the focus of intense research. In this review, we summarize the biochemical properties of RAS proteins, trace down the evolution of RAS signaling and present an overview of the spatio-temporal activation of major RAS isoforms. We further discuss RAS effector pathways, their role in normal and transformed cell physiology and summarize ongoing attempts to interfere with aberrant RAS signaling. Finally, we comment on the role of micro RNAs in modulating RAS expression, contribution of RAS to stem cell function and on high-throughput analyses of RAS signaling networks. PMID- 17428556 TI - Improvement in Doppler alternans in patients with severe heart failure: Levosimendan versus dobutamin. AB - Mechanical alternans, an ominous sign of terminal systolic heart failure, could be indirectly shown by Doppler analysis. On the other hand, Levosimendan, a novel agent, improves contractility in patients with HF. We searched the effect of L versus dobutamin onto Doppler alternans in patients with systolic HF. We found that Levosimendan improves beat-to-beat alteration in patients with systolic HF, demonstrated by pulsed Doppler compared to dobutamin. PMID- 17428557 TI - Culprit only versus complete coronary revascularization during primary PCI. AB - BACKGROUND: Primary percutaneous intervention (PCI) is the treatment of choice for acute ST elevation myocardial infarction. Currently it is recommended to treat only the culprit artery during the acute procedure. Only few reports describe the results of simultaneous non-culprit vessel PCI. The study hypothesizes that complete revascularization during primary PCI can be achieved safely with an improved clinical outcome during the indexed hospitalization. METHODS: One hundred and twenty consecutive patients presented with acute ST elevation myocardial infarction (STEMI) and multivessel coronary stenosis. Ninety five underwent complete revascularization (CR): the culprit artery was opened first followed by dilatation of the other significantly narrowed arteries. Twenty five had culprit only revascularization (COR): the culprit artery only was dilated and the other arteries were left untreated during the primary PCI. RESULTS: Complete revascularization (CR) was associated with reduced incidence of major cardiac events (recurrent ischemia, reinfarction, acute heart failure and in-hospital mortality 16.7 versus 52%, P=0.0001). There was a significant lower rate of recurrent ischemic episodes (4.2% versus 32%, P=0.002), myocardial reinfarction (3.1% versus 16%, P=0.01), reintervention (7.3% versus 32%, P=0.001), acute heart failure (9.4% versus 32%, P=0.01) during the indexed hospitalization and shorter hospitalization (4.4+/-1.27 versus 9.6+/-2.3, P=0.001) in the CR group. Transient renal dysfunction was more common in CR patients (8.4% versus 4% P=0.01). In-hospital and one year mortality were similar between the two groups. CONCLUSION: Multivessel PCI during acute myocardial infarction is feasible and safe. Complete revascularization resulted in an improved acute clinical course. These data support a policy of complete revascularization during primary PCI for STEMI. PMID- 17428558 TI - Early diastolic impairment of diabetic heart: the significance of right ventricle. AB - We read with great interest the study by Karamitsos et al. the methods and interpretation of the results, however, raise several concerns: 1. Are the LVMI higher in patients with type 1 diabetes mellitus than controls? If yes, do patients with type 1 diabetes mellitus and normal LVMI have impaired diastolic function? 2. Is the significant alteration of left and right ventricular diastolic function by tissue Doppler echocardiography in patients with type 1 diabetes mellitus in this study caused by the higher level of glycemia, or is tissue Doppler echocardiography sensitive enough to evaluate early alteration of cardiac function in patients with type 1 diabetes? Therefore, we think that further studies about left and right ventricular systolic and diastolic functions when glycemic control improves and aggravates are needed. PMID- 17428559 TI - Haemodynamic significance of an anomalous right coronary with inter-arterial course assessed with intracoronary pressure measurements during dobutamine challenge. AB - Autopsy studies have associated congenital coronary anomalies with the risk of sudden cardiac death. However, not all patients with anomalous coronary arteries die suddenly. A means of assessing the potential for ischaemia and thus predicting the risk of sudden death in these patients may be necessary for directing treatment. We present the case of a patient with an anomalous right coronary artery originating from the aortic wall above the left sinus of Valsalva, with an anterior inter-arterial course. The haemodynamic significance of this anomaly was assessed using intracoronary pressure-wire measurements at rest and during dobutamine stress. This technique could be a valid means for assessing stress-induced ischaemia and, therefore, by inference, the risk of sudden cardiac death in these patients. PMID- 17428560 TI - A new method for the construction of a mutant library with a predictable occurrence rate using Poisson distribution. AB - A yeast transcriptional activator, Gcn4p, induces the expression of genes that are involved in amino acid and purine biosynthetic pathways under amino acid starvation. Gcn4p has an acidic activation domain in the central region and a bZIP domain in the C-terminus that is divided into the DNA-binding motif and dimerization leucine zipper motif. In order to identify amino acids in the DNA binding motif of Gcn4p which are involved in transcriptional activation, we constructed mutant libraries in the DNA-binding motif through an innovative application of random mutagenesis. Mutant library made by oligonucleotides which were mutated randomly using the Poisson distribution showed that the actual mutation frequency was in good agreement with expected values. This method could save the time and effort to create a mutant library with a predictable mutation frequency. Based on the studies using the mutant libraries constructed by the new method, the specific residues of the DNA-binding domain in Gcn4p appear to be involved in the transcriptional activities on a conserved binding site. PMID- 17428561 TI - The best of both worlds: phase-reset of human EEG alpha activity and additive power contribute to ERP generation. AB - Some authors have proposed that event-related potentials (ERPs) are generated by a neuronal response which is additive to and independent of ongoing activity, others demonstrated that they are generated by partial phase-resetting of ongoing activity. We investigated the relationship between event-related oscillatory activity in the alpha band and prestimulus levels of ongoing alpha activity on ERPs. EEG was recorded from 23 participants performing a visual discrimination task. Individuals were assigned to one of three groups according to the amount of prestimulus total alpha activity, and distinct differences of the event-related EEG dynamics between groups were observed. While all groups exhibited an event related increase in phase-locked (evoked) alpha activity, only individuals with sustained prestimulus alpha activity showed alpha-blocking, that is, a considerable decrease of poststimulus non-phase-locked alpha activity. In contrast, individuals without observable prestimulus total alpha activity showed a concurrent increase of phase-locked and non-phase-locked alpha activity after stimulation. Data from this group seems to be in favor of an additive event related neuronal response without alpha-blocking. However, the dissociable EEG dynamics of total and evoked alpha activities together with a complementary simulation analysis indicated a partial event-related reorganization of ongoing brain activity. We conclude that both partial phase-resetting and partial additive power contribute dynamically to the generation of ERPs. The prestimulus brain state exerts a prominent influence on event-related brain responses. PMID- 17428562 TI - Involvement of substance P and calcitonin gene-related peptide in development and maintenance of neuropathic pain from spinal nerve injury model of rat. AB - Recently, it has been suggested that uninjured primary sensory neurons contribute to neuropathic pain induced by peripheral nerve injury. However, there is lack of evidences of roles of normal pain transmitting substances such as substance P and calcitonin gene-related peptide (CGRP) in neuropathic pain. Whether substance P and CGRP have a role in spinal nerve-injured neuropathic pain model was tested. Male rats were subjected to L5 and L6 spinal nerve transection (SNT), and mechanical hyperalgesia was evaluated by measuring paw withdrawal threshold (PWT). SNT induced a persistent PWT decrease, a sign of neuropathic pain. Lidocaine was soaked on spinal nerves or intrathecally injected 10 min before SNT to block neuronal discharges caused by the injury, and L703,606 (NK1 receptor antagonist) and CGRP8-37 (CGRP receptor antagonist) were intrathecally injected into the rats to block actions of substance P and CGRP released from central nerve terminals in the spinal cord by injury discharges. The treatments with lidocaine, L703,606 and CGRP8-37 delayed the onset of neuropathic pain by 1-4 days, compared with the saline-treated rats. After neuropathic pain was established, intrathecal injections of L703,606 and CGRP8-37 significantly mitigated mechanical hyperalgesia for 20 min. These results suggest that substance P and CGRP are involved in the development and maintenance of neuropathic pain and that these peptides from the central terminals of intact sensory neurons contribute to the maintenance of peripheral nerve injury-induced neuropathic pain. PMID- 17428563 TI - Family-based association study between GRIK2 polymorphisms and autism spectrum disorders in the Korean trios. AB - Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder with strong genetic components. The present study comprises the detection of four single nucleotide polymorphisms (SNPs) in GRIK2 followed by a family-based association analysis of the SNPs in 126 Korean ASD trios by using the transmission disequilibrium test (TDT) and haplotype analysis. We found preferential transmission of the C allele at the rs3213607 (P<0.001) of GRIK2 in ASD and haplotype analysis revealed that one haplotype demonstrated a significant association (P=0.023). These results suggest a potential association between GRIK2 and ASD in the Korean population. PMID- 17428564 TI - Occurrence of Shiga toxin-producing Escherichia coli in a Spanish raw ewe's milk cheese. AB - The aim of the present study was to investigate the occurrence of Escherichia coli O157:H7 and other Shiga toxin-producing E. coli (STEC) in 'Castellano' cheese, a non-cooked and hard or semi-hard Spanish cheese made from ewe's milk. A total of 83 raw milk cheese samples with different ripening times (2.5, 6 and 12 months) were taken at 30 cheese factories. Samples were examined for the presence of STEC using in the first stage the Association of Official Analytical Chemists (AOAC) official method number 997.11, and then, in the second stage, isolates were tested for virulence genes using genotypic (PCR) methods. Three STEC strains were detected in two samples (2.4%) of 'Castellano' cheese, one with 2.5 and the other one with 12 month-ripening period. From those STEC isolates, two were identified as E. coli O14 and the third presented an O-specific polysaccharide not-groupable serologically (ONG). PCR showed that all isolates were characterized by harbouring the Shiga toxin (stx) stx1 gene and by the absence of the genes for stx2, eaeA, and ehxA virulence factors. This study revealed the potential of STEC to survive in long-ripened-hard cheeses. PMID- 17428565 TI - Survival of Shigella sonnei on smooth tomato surfaces, in potato salad and in raw ground beef. AB - This study investigated the survival of a five-strain Shigella sonnei cocktail on smooth tomato surfaces, in potato salad and in raw ground beef. All inocula were resistant to the antibiotic rifampicin to allow simple detection of the target culture among the indigenous microflora of the food samples. Inoculated tomatoes were stored at 13 degrees C/85% relative humidity, the standard holding conditions for mature, green tomatoes. Inoculated potato salad and ground beef samples were stored at 2.5 degrees C and 8.0 degrees C to study the effects of varied refrigerated temperatures. Surviving populations were estimated using a three-tube most probable number (MPN) method. Tryptic soy broth tubes supplemented with 100 ppm rifampicin were inoculated with appropriate dilutions of the recovered inocula and scored for growth after overnight enrichment. S. sonnei populations declined rapidly to undetectable levels (2 days) when dried on smooth surfaces of tomatoes. S. sonnei populations did not decrease in potato salad and ground beef stored at 2.5 degrees C and 8.0 degrees C over the shelf life of the products. PMID- 17428566 TI - Biocontrol of blue and gray mold diseases of pear fruit by integration of antagonistic yeast with salicylic acid. AB - This study was conducted to evaluate the efficacy of the biocontrol yeast Cryptococcus laurentii and salicylic acid (SA) in suppressing the blue and gray mould rots in pear fruit and to explore possible mode of action involved. Our results showed that the combined treatment of pear fruit with C. laurentii with SA at 100 microg ml(-1) resulted in a remarkably improved control of Penicillium expansum and Botrytis cinerea infections, including the pre-inoculated P. expansum, in comparison with the application of C. laurentii or SA alone. The biocontrol yeast C. laurentii proliferated rapidly within the first 24 h of incubation in pear fruit wounds. Although SA at 100 microg ml(-1) neither affected the population growth of C. laurentii nor directly inhibited the blue mold when the inoculation concentrations of P. expansum were above 5 x 10(2) spore per ml in vivo, it induced the fruit resistance to the blue and gray mold rots when the time interval between SA treatment and pathogens inoculation was more than 48 h, being associated with a rapid and strong activation of the peroxidase activity in pear fruit. Thus we assume that SA may be regarded as a secondary defense line in a combination of C. laurentii and SA, which could reinforce the biocontrol efficacy of C. laurentii by induction of the fruit natural resistance. PMID- 17428567 TI - Evidence of natural recombination in classical swine fever virus. AB - Classical swine fever (CSF) virus, one member of the family Flaviviridae is the pathogen of CSF, an economically important and highly contagious disease of pigs. Although homologous recombination has been demonstrated in many other members of the family, it is unknown whether there is recombination in natural populations of CSFV. To detect possible recombination events, we performed a phylogenetic analysis of 25 full-length CSFV strains isolated all over the world. Putative recombinant sequences were identified with the use of SimPlot program. Recombination events were confirmed by bootscaning. A mosaic virus, CSFV 39 (AF407339) isolated in China was found. And its two putative parental-like strains CSFV Shimen (AF333000) and GXWZ02 (AY367767) were identified. Our work revealed that homologous recombination occurred in natural CSFV populations, generating genetic diversity. This would provide some insights for the role homologous recombinant plays in CSFV evolution. PMID- 17428568 TI - Hepatitis C virus molecular clones and their replication capacity in vivo and in cell culture. AB - The hepatitis C virus (HCV) is a positive-strand RNA virus that belongs to the genus Hepacivirus of the family Flaviviridae. The development of a system to propagate this human pathogen in cell culture took more than a decade since the first molecular cloning of the HCV genome. It was a stepwise achievement that began with the construction of the first functional HCV genome with proven in vivo infectivity. It was then followed by the establishment of subgenomic replicons that self-amplify in cultured human hepatoma cells, and culminated in the generation of infectious HCV upon transfection of these cells with a particular molecular HCV clone designated JFH-1. In this review, we will summarize the development and current state of molecular HCV clones and discuss the prospects and implications of the most recent achievements. PMID- 17428569 TI - Effervescent fast-disintegrating bacterial formulation for biological control of rice sheath blight. AB - A lack of effective, easily applied and stable formulation has been a major obstacle to widespread use of biocontrol agents for control of rice sheath blight. In this study, effervescent fast-disintegrating granules containing endospores of Bacillus megaterium were developed for use either by broadcast or spray application. The formulation composed of lactose, polyvinyl pyrrolidone K 30 (PVP, K-30) and effervescent base (citric acid, tartaric acid and sodium bicarbonate). The number of living bacteria in effervescent granules that performed mycelial growth inhibition was in the range of 10(9) CFU/g after 12 months storage at room temperature. The number of viable bacteria after applying into the water and spraying on the rice seedling for 7 days in the greenhouse tests were also satisfactory high (10(9) CFU/g of granules and 10(6) CFU/g of plant, respectively). The scanning electron microscope (SEM) was used to observe bacterial antagonist on the surface of leaf sheath and leaf blade after spraying with formulation. Effervescent formulation applied either broadcasting or spraying reduced incidence of sheath blight disease in the greenhouse experiments. PMID- 17428570 TI - Towards controlled release of BDNF--manufacturing strategies for protein-loaded lipid implants and biocompatibility evaluation in the brain. AB - It was the aim of this study to establish triglyceride matrices as potential carriers for long-term release of brain-derived neurotrophic factor (BDNF), a potential therapeutic for Huntington's disease. First, four different manufacturing strategies were investigated with lysozyme as a model substance: either lyophilized protein was mixed with lipid powder, or suspended in organic solution thereof (s/o). Or else, an aqueous protein solution was dispersed by w/o emulsion in organic lipid solution. Alternatively, a PEG co-lyophilization was performed prior to dispersing solid protein microparticles in organic lipid solution. After removal of the solvent(s), the resulting powder formulations were compressed at 250 N to form mini-cylinders of 2 mm diameter, 2.2 mm height and 7 mg weight. Protein integrity after formulation and release was evaluated from an enzyme activity assay and SDS-PAGE. Confocal microscopy revealed that the resulting distribution of FITC-lysozyme within the matrices depended strongly on the manufacturing method, which had an important impact on matrix performance: matrices with a very fine and homogeneous protein distribution (PEG co lyophilization) continually released protein for 2 months. The other methods did not guarantee a homogeneous distribution and either failed in sustaining release for more than 1 week (powder mixture), completely liberating the loading (s/o dispersion) or preserving protein activity during manufacturing (w/o emulsion, formation of aggregates and 25% activity loss). Based on these results, miniature sized implants of 1 mm diameter, 0.8 mm height and 1 mg weight were successfully loaded by the PEG co-lyophilization method with 2% BDNF and 2% PEG. Release studies in phosphate buffer pH 7.4 at 4 and 37 degrees C revealed a controlled release of either 20 or 60% intact protein over one month as determined by ELISA. SDS-PAGE detected only minor aggregates in the matrix during release at higher temperature. In vivo evaluation of lipid cylinders in the striatum of rat brains revealed a biocompatibility comparable to silicone reference cylinders. PMID- 17428571 TI - Retention of protein complex membership by ancient duplicated gene products in budding yeast. AB - To investigate functional divergence of gene duplicates, we examined the protein protein interactions and coexistence in complexes of paralogs resulting from an ancient whole-genome duplication in yeast. Strikingly, half the surveyed paralog pairs were found to be co-clustered in protein complexes, and were more conserved and highly expressed than non-co-clustered paralogs; however, their discordant expression patterns and conservation rates indicate differential regulation of subfunctionalized paralogs. These results highlight the value of protein complex membership in studying functional divergence among gene duplicates. PMID- 17428572 TI - Combined effects of genetic polymorphisms in six selected genes on lung cancer susceptibility. AB - Various molecular epidemiological studies have been performed to find genetic etiology for lung cancer. Particularly, genetic polymorphisms in NAD(P)H-quinone oxidoreductase (NQO1), cytochrome P450 (CYP)1A1, myeloperoxidase (MPO), glutathione-S-transferase (GST)P1, GSTT1, and GSTM1, and have been suspected to affect lung cancer risk. However, there was no study that examined the combined effects of these genes on lung cancer risk. We studied the combined genetic effects on lung cancer risk in 671 Korean subjects including 318 lung cancer patients and 353 controls. They filled questionnaires, which included lifestyle and childhood- and current environment data. Based on single nucleotide polymorphisms and gene deletions, genetic polymorphisms of the above six genes were determined with PCR-RFLP and TaqMan methods. As results, genetic polymorphisms in the GSTP1, MPO, and CYP1A1 among the genetic factors showed associations with lung cancer risk. The reference, which is supposed to have the lowest risk for cancer, was subjects who were homozygous wild type of the GSTP1 and CYP1A1 and had the MPO- mutant allele. After combination study of the three gene-polymorphism, the subjects who were most different with the reference, i.e. had the mutant allele of the GSTP1 and CYP1A1 and homozygous wild type of the MPO, showed approximately 5-fold-higher risk for lung cancer than the reference (95% CI, 2.05-12.05). Therefore, our study suggests that the combination of the GSTP1, MPO, and CYP1A1 variations affects susceptibility to lung cancer. PMID- 17428573 TI - Evaluation of groundwater flow patterns around a dual-screened groundwater circulation well. AB - Dual-screened groundwater circulation wells (GCWs) can be used to remove contaminant mass and to mix reagents in situ. GCWs are so named because they force water in a circular pattern between injection and extraction screens. The radial extent, flux and direction of the effective flow of this circulation cell are difficult to measure or predict. The objective of this study is to develop a robust protocol for assessing GCW performance. To accomplish this, groundwater flow patterns surrounding a GCW are assessed using a suite of tools and data, including: hydraulic head, in situ flow velocity, measured hydraulic conductivity data from core samples, chemical tracer tests, contaminant distribution data, and numerical flow and transport models. The hydraulic head data show patterns that are consistent with pumping on a dual-screened well, however, many of the observed changes are smaller than expected. In situ thermal perturbation flow sensors successfully measured horizontal flow, but vertical flow could not be determined with sufficient accuracy to be useful in mapping flow patterns. Two types of chemical tracer tests were utilized at the site and showed that much of the flow occurs within a few meters of the GCW. Flow patterns were also assessed based on changes in contaminant (trichloroethylene, TCE) concentrations over time. The TCE data clearly showed treated water moving away from the GCW at shallow and intermediate depths, but the circulation of that water back to the well, except very close to the well, was less clear. Detailed vertical and horizontal hydraulic conductivities were measured on 0.3 m-long sections from a continuous core from the GCW installation borehole. The measured vertical and horizontal hydraulic conductivity data were used to construct numerical flow and transport models, the results of which were compared to the head, velocity and concentration data. Taken together, the field data and modeling present a fairly consistent picture of flow and transport around the GCW. However, the time and expense associated with conducting all of those tests would be prohibitive for most sites. As a consequence, a sequential protocol for GCW characterization is presented here in which the number of tools used can be adjusted to meet the needs of individual sites. While not perfect, we believe that this approach represents the most efficient means for evaluating GCW performance. PMID- 17428574 TI - Individual differences in food perceptions and calorie estimation: an examination of dieting status, weight, and gender. AB - People frequently place foods into "health" or "diet" categories. This study examined whether (1) evaluations of "healthiness/unhealthiness" influence "caloric" estimation accuracy, (2) people evaluate foods for "healthiness/unhealthiness" or "weight gain/loss" differently, and (3) food evaluations differ by gender, diet status, and weight. Also, undergraduate dieters attempting to lose weight on their own were compared to obese weight loss program participants. Undergraduate students (N=101) rated eight "healthy" and "unhealthy" foods on perceived "healthiness/unhealthiness," "weight loss/gain capacity" and "caloric" content. Open-ended questions inquiring why a food was "healthy/unhealthy" or would "contribute to weight gain/loss" were coded into independent food categories (e.g., high fat). Results indicate that calories were systematically underestimated in healthy/weight loss foods, while they were systematically overestimated in unhealthy/weight gain foods. Dieters were more accurate at estimating "calories" of healthy foods and more attentive to the foods' fat, "calorie", and sugar content than non-dieters. Overweight participants commented more on fat and sugar content than normal weight participants. Undergraduate dieters used fewer categories for evaluating foods than weight loss program participants. Individual difference characteristics, such as diet-status, weight, and gender, influence people's perceptions of foods' healthiness or capacity to influence weight, and in some instances systematically bias their estimates of the caloric content of foods. PMID- 17428575 TI - Fungal contamination of hospital healthcare workers' overalls. PMID- 17428576 TI - Outbreak of Enterobacter hormaechei septicaemia in newborns caused by contaminated parenteral nutrition in Brazil. PMID- 17428577 TI - Long-term effect of a 1.5 minute surgical hand rub with a propanol-based product on the resident hand flora. PMID- 17428578 TI - The problem with Penicillium. PMID- 17428579 TI - Isolation of vancomycin-resistant Staphylococcus aureus in a teaching hospital in Tehran. PMID- 17428580 TI - Increased levels of C-type natriuretic peptide in patients with idiopathic left ventricular dysfunction. AB - C-type natriuretic peptide (CNP) is expressed in the vascular endothelium. It is not known whether CNP is specifically increased in patients with idiopathic left ventricular systolic dysfunction (ILVDys) with or without overt heart failure, and whether in these patients it is related with indicators of myocardial and/or endothelial/microvascular impairment. We determined plasma CNP levels in 51 ILVDys and in 60 controls. We observed a significant increase in patients with (7.0+/-0.9 pg/ml) or without (6.1+/-0.53 pg/ml) overt heart failure (p<0.001) in respect to controls (2.5+/-0.12 pg/ml). CNP was significantly correlated with LVEF (p<0.001), end-diastolic dimension (p<0.05), ANP (p<0.001) and BNP (p<0.001), interleukin-6 (p<0.001), total cholesterol (p<0.05), low-density lipoprotein (p=0.05), ratio total cholesterol/ high-density lipoprotein (p=0.05) and, in a subgroup of patients, with abnormal vasodilating capacity of the coronary microcirculation. In conclusion, CNP is activated in patients with LV dysfunction but without coronary artery disease, independently of the presence of overt heart failure and in tune with the extent of myocardial functional involvement. In these patients CNP is also related with both systemic and coronary indicators of endothelial/microvascular damage. PMID- 17428581 TI - Diagnostic performance of a CSF-biomarker panel in autopsy-confirmed dementia. AB - To establish diagnostic performance of the cerebrospinal fluid (CSF) biomarkers beta-amyloid peptide (Abeta(1-42)), total tau-protein (T-tau) and tau phosphorylated at threonine 181 (P-tau(181P)) compared to clinical diagnosis, biomarker levels were determined in CSF samples from 100 autopsy-confirmed dementia and 100 control subjects. As the control and dementia groups were not age-matched and given the significant associations of biomarker concentrations with age in controls, age-corrected biomarker concentrations were calculated. New models were constructed by means of logistic regression. Using all biomarkers, dementia could be discriminated from controls (sensitivity (S)=86%, specificity (Sp)=89%). T-tau and Abeta(1-42) optimally discriminated Alzheimer's disease (AD) from other dementias (NONAD) and controls (S=90%, Sp=89%). AD was optimally discriminated from NONAD using P-tau(181P) and Abeta(1-42) (S=80%, Sp=93%). Diagnostic accuracy of the latter model (82.7%) was comparable to clinical diagnostic accuracy (81.6%) that was based on a whole clinical work-up (including imaging). Using this model, in cases with clinically doubtful diagnoses, a correct diagnosis would have been established in 4/6 autopsy-confirmed AD and 3/3 autopsy-confirmed NONAD cases. The value of biomarkers in differential dementia diagnosis was shown, using pathological diagnosis as a reference. New models have been developed, achieving sensitivity, specificity and diagnostic accuracy levels, consistently exceeding 80%. PMID- 17428582 TI - Motor memory preservation in aged monkeys mirrors that of aged humans on a similar task. AB - We studied long-term motor memory preservation in rhesus monkeys tested on a task similar to that employed in humans. First, motor speed and rate of motor decline was measured in 23 animals ranging from 4 to 26 years old. The task for the animals consisted of removing a food reward from a curved rod within the inner chamber of an automated panel. Young animals performed twice as fast as the aged animals. Second, young (n=6) and aged (n=10) animals were re-tested 1 year later on the same task with no intervening practice. We anticipated a decline in motor speed of 144 ms/year, instead the average performance time recorded during the repeat session improved significantly by 17% in the aged animals. This finding mirrors that of a longitudinal study conducted in humans using a similar test panel and supports that, while initial performance times of a novel motor task decline with age, motor memory traces are preserved over an extended time interval, even without continued practice. The data also support that the rhesus monkey could be used as a model to study the mechanisms by which long-term retention of motor memory occurs in aging. PMID- 17428583 TI - Solvatochromism, DNA binding, antitumor activity and molecular modeling study of mixed-ligand copper(II) complexes containing the bulky ligand: bis[N-(p tolyl)imino]acenaphthene. AB - Four mixed-ligand copper(II) complexes of the nitrogen ligand bis[N-(p tolyl)imino]acenaphthene 1 (p-Tol-BIAN). These complexes, namely [Cu(p-Tol BIAN)2](ClO4)2 2, [Cu(p-Tol-BIAN)(acac)](ClO4) 3, [Cu(p-Tol-BIAN)Cl2] 4 and [Cu(p Tol-BIAN)(AcOH)(2)](ClO4)2 5, were prepared and characterized. Solvatochromism of the novel copper complexes in various solvents has been studied. Molecular mechanics (MM+) and molecular dynamic simulations have been performed to learn more about the solvatochromic behaviour and the DNA binding affinity of these complexes. PMID- 17428584 TI - Implementation and evaluation of a pilot education programme in colorectal cancer management for nurses in Scotland. AB - This paper describes the results of an evaluation of a nursing education programme for nurses caring for patients with colorectal cancer. A sample of 67 registered nurses from 6 location sites within the 3 Regional Cancer Networks in Scotland were recruited to the pilot. The programme was adapted from an evidence based education manual for nurses in the management of colorectal cancer developed by the European Oncology Nursing Society as part of a Nursing in Colorectal Cancer Initiative (NICCI) [Hawthorn, J., Redmond, K., 1999. A Guide to Colorectal Cancer. AstraZeneca Oncology, UK]. The format for evaluating the programme was based on the TELER method of treatment evaluation [Le Roux, A.A., 1995. TELER: the concept. Physiotherapy 79 (11), 755-758] that had previously been developed along side the training manual [Grocott, P., Richardson, A., Ambaum, B., Kearney, N., Redmond, K, 2001a. Nursing in colorectal cancer initiative--the audit phase. Part 1. Development of the audit tool. European Journal of Oncology Nursing 5 (2), 100-111; Grocott, P., Richardson, A., Ambaum, B., Kearney, N., Redmond, K., 2001b. Nursing in colorectal cancer initiative: the audit phase. Part 2. Content validity of the audit tool and implications of the standards set for clinical practice. European Journal of Oncology Nursing 5 (3), 165-173] for cytotoxic chemotherapy to provide the NICCI Audit Tool (Le Roux, 2003). This model was developed further in the current study to include the domains of: Disease, Diagnosis and Staging, Treatment, Nursing Issues and General Issues. Data were analysed descriptively and are discussed. Overall the results from this study demonstrate a statistically significant improvement in disease related knowledge (p=<0.001) and in the best practice statements for nursing issues (p=<0.001) and general issues (including attitudes) (p=0.023) that were maintained at four months post completion of the course. PMID- 17428585 TI - A novel RNA silencing vector to improve antigen expression and stability in Chinese hamster ovary cells. AB - Chinese hamster ovary (CHO) cells and dihydrofolate reductase (dhfr)/methotrexate (MTX) gene amplification system are routinely used to generate stable producer CHO cell clones in biopharmaceutical industries. The present study proposes a novel method by the co-amplification of the silencing vector targeted to dhfr gene for improvements of selecting high-producing clones in dhfr-deficient and wild-type CHO cells. Using the silencing vector also resulted in improving the stability of the recombinant protein expression in the absence of MTX in the CHO/dhFr(-) and wild-type CHO cells. This new method is proposed to generate highly expressed stable cell clones of both dhfr-deficient and wild-type CHO cells for recombinant antigen production. Utilization of the silencing vector designed in this study can improve antigen expression through dhfr-directed gene amplification in other dhfr-competent cell lines for vaccine development. PMID- 17428586 TI - A Vibrio cholerae serogroup O1 vaccine candidate against CTX ET Phi infection. AB - Cholera is a severe diarrheal disease that may spread rapidly. Vaccination is considered a valid measure against it. We developed a new vaccine candidate, IEM109, against Vibrio cholerae. To generate this candidate, a chromosomal fragment containing the TLC element, attB of the CTX Phi integration site, and RTX cluster responsible for the cytotoxic activity for mammalian cells was deleted through homologous recombination from the previously described El Tor biotype, IEM101. The protective genes ctxB and rstR, which establish resistance to CTX Phi infections, were inserted into that same location on the chromosome of IEM109 to enhance the safety and genetic stability of the vaccine candidate and to prevent horizontal gene transfer. In in vivo tests, cell cultures showed that the cytotoxic effect of IEM109 on Hep-2 was negative. Furthermore, the infection rate of El Tor biotype CTX Phi to that of IEM109 in the rabbit intestine is 3000 fold lower than that of IEM101. Intraintestinal vaccination of rabbits with a single dose of IEM109 elicits high titers of anti-CTB IgG and vibriocidal antibodies. When challenged with 0.5-2 microg CT and 10(5) to 10(8)CFU of four wild toxigenic strains of different biotypes and serogroups, IEM109 conferred full protection. Thus, IEM109 is a stable vaccine candidate that evokes not only antitoxic and vibriocidal immunities, but also resistance to the El Tor biotype CTX Phi infection. PMID- 17428587 TI - Conjugating recombinant proteins to Pseudomonas aeruginosa ExoProtein A: a strategy for enhancing immunogenicity of malaria vaccine candidates. AB - Conjugation of polysaccharides to carrier proteins has been a successful approach for producing safe and effective vaccines. In an attempt to increase the immunogenicity of two malarial vaccine candidate proteins of Plasmodium falciparum, apical membrane antigen 1 (AMA1) to a blood stage vaccine candidate and surface protein 25 (Pfs25) a mosquito stage vaccine candidate, were each independently chemically conjugated to the mutant, nontoxic Pseudomonas aeruginosa ExoProtein A (rEPA). AMA1 is a large (66kD) relatively good immunogen in mice; Pfs25 is a poorly immunogenic protein when presented on alum to mice. Mice were immunized on days 0 and 28 with AMA1- or Pfs25-rEPA conjugates or unconjugated AMA1 or Pfs25, all formulated on Alhydrogel. Remarkably, sera from mice 14 days after the second immunization with Pfs25-rEPA conjugates displayed over a 1000-fold higher antibody titers as compared to unconjugated Pfs25. In contrast, AMA1 conjugated under the same conditions induced only a three-fold increase in antibody titers. When tested for functional activity, antibodies elicited by the AMA1-rEPA inhibited invasion of erythrocytes by blood-stage parasites and antibodies elicited by the Pfs25-rEPA conjugates blocked the development of the sexual stage parasites in the mosquito midgut. These results demonstrate that conjugation to rEPA induces a marked improvement in the antibody titer in mice for the poor immunogen (Pfs25) and for the larger protein (AMA1). These conjugates now need to be tested in humans to determine if mice are predictive of the response in humans. PMID- 17428588 TI - Scientific consultation on immunological correlates of protection induced by dengue vaccines report from a meeting held at the World Health Organization 17-18 November 2005. AB - Several dengue vaccine candidates have been evaluated in early clinical phase, and some are scheduled for efficacy testing in population-based studies. Given the advancements in dengue vaccine development, there is an increased interest in identifying immunological correlates of protection for these vaccines in order to facilitate their evaluation, further refinement, production and registration. To this end, the WHO Initiative for Vaccine Research (IVR) convened a consultation on primary and secondary immunological correlates of protection induced by dengue vaccines. The meeting was held on the 17th and 18th of November, 2005 at WHO headquarters in Geneva. The consultation was a first dedicated review of the available data in support of establishing correlates. It is concluded that it is not yet possible to define one specific set of correlates, the consultation concluded in recommendations that should help to gather the missing evidence in conjunction with future vaccine trials. PMID- 17428589 TI - Radioactivity and radiological risk associated with effluent sediment containing technologically enhanced naturally occurring radioactive materials in amang (tin tailings) processing industry. AB - The processing of amang, or tin tailings, for valuable minerals has been shown to technologically enhance NORM and this has stirred significant radiological safety and health concerns among Malaysia's regulatory authority. A growing radiological concern is now focused on the amang effluent containing NORM in recycling ponds, since these ponds may be reclaimed for future residential developments. A study was carried out to assess the radiological risk associated with amang processing and the accumulated effluent in the recycling ponds. Twenty-six sediment samples from the recycling ponds of two amang plants in the states of Selangor and Perak, Malaysia, were collected and analyzed. The maximum activity concentrations of (238)U, (226)Ra, (232)Th and (40)K recorded in sediments from these ponds were higher than Malaysia's and the world's natural highest. Correspondingly, the mean radium equivalent activity concentration indices, Ra(eq), and gamma radiation representative level index, I(gammar), were higher than the world's average. The enhancement of NORM in effluent sediments as a consequence of amang processing, and the use of a closed water management recycling system created Effective Dose Rates, E (nSv h(-1)), that signal potential environmental radiological risks in these ponds, should they be reclaimed for future land use. PMID- 17428590 TI - The current content of artificial radionuclides in the water of the Tobol-Irtysh river system (from the mouth of the Iset River to the confluence with the Ob River). AB - Data on content of (90)Sr, (137)Cs, (239,240)Pu and (3)H in water of the Tobol Irtysh part of the Techa-Iset-Tobol-Irtysh-Ob river system (through which the "Mayak" PA radioactive wastes are transported) are presented and discussed. The data were received in 2004-2005 under the ISTC project on radioecological monitoring of the Tobol and Irtysh rivers. Monthly observations of (137)Cs, (90)Sr and (3)H content in water in the area of the Tobol and Irtysh confluence have been conducted starting from May 2004. To obtain information on the investigated river system as a whole, the radioecological survey of the Tobol and Irtysh rivers at the section from the mouth of the Iset River to the confluence with the Ob River was carried out in 2004. It is shown that the impact of "Mayak" PA waste transport by (90)Sr is distinctly traced as far as the area of the Irtysh and Ob confluence. PMID- 17428591 TI - Measurements of short-lived cosmic-ray-produced radionuclides in rainwater. AB - Cosmic-ray-produced (CP) nuclides with half-lives shorter than 21h were measured in rainwater by ultra-low-background gamma spectrometry at Ogoya Underground Laboratory. As levels of CP nuclides are extremely low and their half-lives are very short, quick sampling methods for a large volume of rainwater and rapid chemical separations by ion exchange method were developed. The nuclides measured were short-lived (24)Na, (28)Mg, (38)S, (38)Cl, (39)Cl, as well as nuclides with longer half-lives (7)Be and (22)Na. The number of atoms of CP nuclides in rainwater were evaluated to range from 30 to 1500L(-1) for (24)Na (n=16, mean; 520 [6.7mBqL(-1)]), 80 to 600L(-1) for (28)Mg (n=13, mean; 260 [2.4mBqL(-1)]), 400 to 1900L(-1) for (39)Cl (n=6, mean; 1200 [250mBqL(-1)]), 1x10(6) to 4x10(7)L( 1) for (7)Be (n=16, mean; 7x10(6) [1.05BqL(-1)]) and 2x10(3) to 1x10(5)L(-1) for (22)Na (n=9, mean; 2x10(4) [0.2mBqL(-1)]). PMID- 17428592 TI - Multiple stressor effects of radiation and metals in salmon (Salmo salar). AB - These experiments were designed to look at the cellular effects in key organs in Atlantic salmon (Salmo salar) after exposure in vivo to radiation and subtoxic levels of aluminum (Al) and cadmium (Cd), alone or in combination. Salmon (25g) were exposed to a single 0.5Gy dose of gamma-irradiation in water containing Cd, Al or Cd+Al. Three fish per group were sacrificed after 1h and the liver, pronephros, fin and gill of each was dissected. Small explants of each tissue were set up. After 2 days, the culture medium was harvested and filtered then placed on a reporter cell line for determination of stress signal activity (bystander effects). Radiation in combination with Cd and/or Al, caused bystander effects in tissues harvested from in vivo exposed salmon. The effects vary between different organs and are not consistently additive or synergistic for a given treatment. Tissue type appears to be critical. Liver cultures produce a toxic factor which is lethal to reporter cells, and therefore no liver data could be obtained. It is hoped that this stress signal response will prove to be a useful indicator of environmental stress in species inhabiting aquatic ecosystems. PMID- 17428593 TI - Metal contamination of Posidonia oceanica meadows along the Corsican coastline (Mediterranean). AB - The aim of this study is to determine metal (Cd, Co, Cr, Hg, Ni, Pb) concentrations in Posidonia oceanica tissues along the Corsican coastline. The results show that except for Cr, all the metals are preferentially accumulated in the blades; this is particularly interesting as it means that future metal analyses may be carried out only on the blades avoiding thus the removal of the shoots. Moreover, they show that metal concentrations may reflect the "background noise" of the Mediterranean Sea. Station 15 (Canari) can however be distinguished from the others due to its high Co, Cr and Ni concentrations. This result may be related to the presence of a previous asbestos mine, located near this station. Therefore, this study reinforces the usefulness and the relevance of Posidonia oceanica as a tracer of spatial metal contamination and as an interesting tool for water quality evaluation. PMID- 17428594 TI - Future climate scenarios and rainfall--runoff modelling in the Upper Gallego catchment (Spain). AB - Global climate change may have large impacts on water supplies, drought or flood frequencies and magnitudes in local and regional hydrologic systems. Water authorities therefore rely on computer models for quantitative impact prediction. In this study we present kernel-based learning machine river flow models for the Upper Gallego catchment of the Ebro basin. Different learning machines were calibrated using daily gauge data. The models posed two major challenges: (1) estimation of the rainfall-runoff transfer function from the available time series is complicated by anthropogenic regulation and mountainous terrain and (2) the river flow model is weak when only climate data are used, but additional antecedent flow data seemed to lead to delayed peak flow estimation. These types of models, together with the presented downscaled climate scenarios, can be used for climate change impact assessment in the Gallego, which is important for the future management of the system. PMID- 17428595 TI - Is an adjusted rhizosphere-based method valid for field assessment of metal phytoavailability? Application to non-contaminated soils. AB - Previously recommended rhizosphere-based method (RHIZO) applied to moist rhizosphere soils was integrated with moist bulk soils, and termed adjusted-RHIZO method (A-RHIZO). The A-RHIZO and RHIZO methods were systematically compared with EDTA, DTPA, CaCl2 and the first step of the Community Bureau of Reference (BCR1) methods for assessing metal phytoavailability under field conditions. Results suggested that moist bulk soils are equally suited or even better than rhizosphere soils to estimate metal phytoavailability. The A-RHIZO method was preferred to other methods for predicting the phytoavailability of Ni, Cu, Zn, Cd, Pb and Mn to wheat roots with correlation coefficients of 0.730 (P<0.001), 0.854 (P<0.001), 0.887 (P<0.001), 0.739 (P<0.001), 0.725 (P<0.001) and 0.469 (P<0.05), respectively. When including soil properties, other extraction methods were also able to predict phytoavailability reasonably well for some metals. Soil pH, organic matter and Fe-Mn oxide contents, and cation-exchange capacity mostly influenced the extraction and phytoavailability of metals. PMID- 17428596 TI - The 'world of yoga': the production and reproduction of therapeutic landscapes. AB - Yoga is becoming more and more fashionable all around the world. This activity, partly considered as therapeutic, reveals contemporary ways of producing global practices. Via a questionnaire completed by individuals at yoga centres in India and France, the paper analyses this phenomenon using the concept of therapeutic landscapes. Furthermore, it examines how these therapeutic landscapes are influenced by globalisation. Bringing together the concepts of therapeutic landscapes, globalisation and the practice of yoga, the paper analyses the production and reproduction of yogic therapeutic landscapes in the world's space. Constituted of natural physical elements and built structures, these therapeutic landscapes are also strongly linked to emotional qualities and intimate feelings evoked by the place and related to health and well-being. The paper also underlines that the emergence of internationally recognised therapeutic landscapes demands a cultural and social geographical approach that could help to examine the repercussions of local and global phenomena on therapeutic landscapes. Indeed, very often, yogis, all around the world, refer to ideal places where they rarely go. Consequently, they create an imagined therapeutic environment that the paper describes. PMID- 17428597 TI - "I wanted to do a good job": experiences of 'becoming a mother' and breastfeeding in mothers of very preterm infants after discharge from a neonatal unit. AB - In mothers of preterm infants, the process of becoming a mother is initiated in a public and medical environment, in which the mothers become dependent on the benevolence and support of the staff. This setting and an experience of insecure social bonds impair the ability to become a mother during the infant's stay at the neonatal unit (NU), and breastfeeding may become a duty and not be mutually satisfying. Studies on how women experience becoming a mother and breastfeeding after the infant's discharge are sparse and this question is addressed in the present grounded theory study. Twenty five mothers, whose very preterm infants had received care in seven NUs in Sweden, were interviewed once, 1-12 months after discharge. We propose a model to increase understanding of the process of becoming a mother and breastfeeding, after the infant's discharge from the NU. The mother's emotional expressions in this process showed pendular swings from feeling emotionally exhausted to feeling relieved, from experiencing an insecure to a secure bond, and from regarding breastfeeding as being non-reciprocal to being reciprocal. Unresolved grief, the institutional authority at the NU and experiences of shame were three of the central barriers to a secure and reciprocal relationship. The pendular changes give us a deeper understanding of the variations in both attachment and attunement. Perhaps the negative extremes are more prominent among these mothers on account of their infant's illness and their NU experiences. If our proposed model is valid, it is vital that these findings are considered by those involved in the short- and long-term care in order to support the mothers to establish a secure bond, comprising both attachment and attunement. PMID- 17428598 TI - Maternal and child health problems: the inter-generational consequences of early maternal aggression and withdrawal. AB - The purpose of this study was to determine the impact of childhood aggression and social withdrawal on adult health, and to examine the transfer of health-risk to offspring. Previous studies have linked aggression to poor health habits, whereas social withdrawal has been linked to increased symptom reporting. Little research has attempted to examine the effects of parents' childhood behaviour problems on health in the next generation. Seventy-four parents from disadvantaged neighbourhoods in Montreal (roughly half of whom were identified in childhood as being highly aggressive and/or withdrawn) and their offspring aged 9-11-years old participated in this prospective, longitudinal study. Health histories of mothers and target children were taken during structured home interviews. Regression analyses tested the relationship between parents' behaviour problems in childhood and subsequent health outcomes in both generations. Mothers who were high on both aggression and withdrawal in childhood showed the earliest signs of health problems. Maternal childhood aggression predicted lower self-rated health and some current health problems, whereas maternal childhood withdrawal predicted more somatizing symptoms such as frequent headaches. Offspring of socially withdrawn mothers or fathers were more likely to be prescribed medication for respiratory problems and Ritalin, despite no matching maternal reports of higher incidence of behavior or respiratory problems. In summary, there were distinct health patterns for families of individuals who are highly aggressive and socially withdrawn in childhood, with implications for transfer of poor health habits to the next generation. PMID- 17428599 TI - Heat-shock protein-70 genes and response to antidepressants in major depression. AB - In the search of predictors of antidepressant efficacy, much interest has recently focused on pro-inflammatory proteins, as they were found to be elevated during major depressives states and decreased by antidepressant drugs. In the present paper we investigated the role of the genes coding for heat-shock-70 family proteins, recently hypothesized to be activated by antidepressants and thus mediate the reduction of pro-inflammatory cytosines. One hundred and forty two hospitalised patients, affected by major depression and treated with antidepressants drugs for a major depressive episode were evaluated for depressive severity at the baseline and at the discharge and genotyped for five SNPs within the genes HSPA1L, HSPA1A and HSPA1B. Markers were not individually associated with symptom severity after treatment. Instead, we found a three markers haplotype, including SNPs within HSPA1L and HSPA1A, associated with a poorer response to antidepressant treatment (p=0.005). Single markers as well as haplotypes were not associated with other clinical features. In conclusion, genetic variants within the genes coding for HSP-70 family proteins may affect the action of antidepressants and thus their therapeutic efficacy. PMID- 17428600 TI - Long-term follow-up of composite resin restorations with self-etching adhesives. AB - OBJECTIVE: To test the hypothesis, derived from a previous short-term (7-day) assessment, that the absence of conventional pulp protection is not responsible for long-term pulp complications of composite resin restorations with self etching adhesives. METHODS: All 150 patients who received the restorations with self-etching adhesives were recalled at least 2 years after the placement of restorations. Of the 47 patients (31%) who responded, 106 restorations aged from 2.2 to 6.5 years were examined for tooth sensitivity and pulp vitality regarding long-term pulp complications. The results were subjected to a multivariable logistic regression analysis with regard to cavity depth, provision of conventional pulp protection and short-term pulp complications. RESULTS: No positive cases were found in the assessment of tooth sensitivity. Four restorations (3.7%) made in deep cavities with conventional pulp protection resulted in pulpectomy due to inflammation, of which three cases presented short term pulp complications. The 95% confidence intervals for the odds ratios estimated by the multivariable logistic regression analysis were (1.54, infinity) for cavity depth (1.50, infinity) for short-term pulp complications and (0.02, infinity) for conventional pulp protection. Namely, the last variable had no significant effect on long-term pulp complications and thus the hypothesis was verified. CONCLUSIONS: The absence of conventional pulp protection was not responsible for long-term pulp complications even in deep cavities with the use of self-etching adhesives. A deep cavity and the existence of short-term pulp complications were two critical predictors for the occurrence of long-term pulp complications. PMID- 17428601 TI - The N-formyl peptide receptors and the anaphylatoxin C5a receptors: an overview. AB - Leukocyte recruitment to sites of inflammation and infection is dependent on the presence of a gradient of locally produced chemotactic factors. This review is focused on current knowledge about the activation and regulation of chemoattractant receptors. Emphasis is placed on the members of the N-formyl peptide receptor family, namely FPR (N-formyl peptide receptor), FPRL1 (FPR like 1) and FPRL2 (FPR like-2), and the complement fragment C5a receptors (C5aR and C5L2). Upon chemoattractant binding, the receptors transduce an activation signal through a G protein-dependent pathway, leading to biochemical responses that contribute to physiological defense against bacterial infection and tissue damage. C5aR, and the members of the FPR family that were previously thought to be restricted to phagocytes proved to have a much broader spectrum of cell expression. In addition to N-formylated peptides, numerous unrelated ligands were recently found to interact with FPR and FPRL1. Novel agonists include both pathogen- and host-derived components, and synthetic peptides. Antagonistic molecules have been identified that exhibit limited receptor specificity. How distinct ligands can both induce different biological responses and produce different modes of receptor activation and unique sets of cellular responses are discussed. Cell responses to chemoattractants are tightly regulated at the level of the receptors. This review describes in detail the regulation of receptor signalling and the multi-step process of receptor inactivation. New concepts, such as receptor oligomerization and receptor clustering, are considered. Although FPR, FPRL1 and C5aR trigger similar biological functions and undergo a rapid chemoattractant-mediated phosphorylation, they appear to be differentially regulated and experience different intracellular fates. PMID- 17428602 TI - Influence of air-purifying respirators on the simulated first response emergency treatment of CBRN victims. AB - AIM: Medical first responders and emergency room personnel potentially are threatened by exposure to primary or secondary intoxication by chemical, biological, radiological or nuclear (CBRN) substances. The impact of personal respiratory protection by air-purifying respirators on the performance of resuscitation requires evaluation. This will help to improve major incident planning and measures for protecting medical staff. METHODS: We investigated the influence of two air-purifying respirator designs on the resuscitation of simulated CBRN victims. Fourteen UK paramedics followed a standardised resuscitation algorithm, either unprotected or wearing a bi-ocular and a panoramic visor respirator in a randomised crossover design. Treatment times and wearer comfort was determined and compared. RESULTS: We did not find any difference in treatment times between the groups wearing respiratory protection and the controls (189+/-8.3s for the controls, 191+/-9.5s for the panoramic visor mask and 206+/-9.1s for the bi-ocular respirator [mean+/-S.E.M.]). Tracheal intubation appeared to be the most time consuming task. In a questionnaire, volunteers were of the opinion that orientation whilst wearing the respirator with the panoramic visor was better compared to the bi-ocular one (85% versus 15%). With respect to the fit, the majority (79%) rated the bi-ocular respirator as more comfortable. CONCLUSIONS: Modern personal respiratory protection has only a negligible effect in the delay on the short term treatment during well defined simulated scenarios with a single CBRN casualty. Furthermore, air-purifying respirators with panoramic visors seem to allow a better orientation for medical first responders during simulated resuscitation. PMID- 17428603 TI - Structure and functions of the human amyloid precursor protein: the whole is more than the sum of its parts. AB - The amyloid precursor protein (APP) is a transmembrane protein that plays major roles in the regulation of several important cellular functions, especially in the nervous system, where it is involved in synaptogenesis and synaptic plasticity. The secreted extracellular domain of APP, sAPPalpha, acts as a growth factor for many types of cells and promotes neuritogenesis in post-mitotic neurons. Alternative proteolytic processing of APP releases potentially neurotoxic species, including the amyloid-beta (Abeta) peptide that is centrally implicated in the pathogenesis of Alzheimer's disease (AD). Reinforcing this biochemical link to neuronal dysfunction and neurodegeneration, APP is also genetically linked to AD. In this review, we discuss the biological functions of APP in the context of tissue morphogenesis and restructuring, where APP appears to play significant roles both as a contact receptor and as a diffusible factor. Structural investigation of APP, which is necessary for a deeper understanding of its roles at a molecular level, has also been advancing rapidly. We summarize recent progress in the determination of the structure of isolated APP fragments and of the conformations of full-length sAPPalpha, in both monomeric and dimeric states. The potential role of APP dimerization for the regulation of its biological functions is also discussed. PMID- 17428604 TI - Use of an activated carbon from antibiotic waste for the removal of Hg(II) from aqueous solution. AB - Porous carbon has been prepared from waste antibiotic material by a chemical activation method using K(2)CO(3) as an activating reagent. Carbon was studied systematically by the adsorption of nitrogen and iodine. It was found that the process parameters such as activation temperature and activation time are crucial for preparing high-quality activated carbon. The proper choice of the preparation conditions allows to produce microporous activated carbon with a micropore volume up to 0.492 cm(3)/g and a BET surface area of 1260 m(2)/g. Adsorption of mercury(II) from an aqueous solution on antibiotic carbon was investigated under the varying conditions of agitation time, metal ion concentration and pH. The adsorption capacity of the carbon is 129 mg/g. PMID- 17428605 TI - Decolorization of C.I. Acid Yellow 23 solution by electrocoagulation process: investigation of operational parameters and evaluation of specific electrical energy consumption (SEEC). AB - This study investigates the evaluation of specific electrical energy consumption (SEEC) and the influence of operating parameters on the color removal efficiency of a dye solution containing C.I. Acid Yellow 23 by electrocoagulation process. Firstly, the operational parameters including current density, initial dye concentration, initial pH and time of electrolysis were optimized. Then the effects of the conductivity, the interelectrode distance and the area of cross section of the electrodes on specific electrical energy consumption (SEEC) were studied under the optimum conditions. Our results indicated that for a solution of 50mg/l C.I. Acid Yellow 23, almost 98% color and 69% chemical oxygen demand (COD) were removed, when the pH was about 6, the time of electrolysis was 5min and the current density was approximately 112.5A/m(2). In addition, the results of our study revealed that when the conductivity and area of cross-section of the electrodes increased and interelectrode distance decreased, the cell voltage and specific electrical energy consumption would be decreased. PMID- 17428606 TI - Electro-analysis of energetic materials. AB - Cyclic voltammetric studies of triaminoguanidine nitrate (TAGN), 3,3'-hydrazino bis(bis[6,6'-(3,5-dimethylpyrazol-lyl])-1,2,4,5-tetrazine (HBPT), 4,6 dinitrobenzofuroxan (DNBF) and 3,3'-diamino-4,4'-azoxyfurazan (DAAF) were carried out at different pH conditions in 50% aqueous acetonitrile using glassy carbon electrode. Optimum pH was selected for individual compounds. Influence of scan rate and concentration on the voltammetric response were studied in optimum pH. The number of electron transferred was determined by controlled potential coulometry. All compounds undergo diffusion controlled electrochemical reaction. Based on cyclic voltammetric results, differential pulse and square wave voltammetric methods have been developed for the analytical determination. Instrumental parameters such as initial scan potential, amplitude, pulse increment, pulse period, pulse width and frequency were studied. Optimum experimental conditions for each compound were obtained. After fixing optimum conditions, the effect of concentration was studied and calibration plot was arrived. These plots can be used to determine the traces of the above said four energetic materials. PMID- 17428607 TI - Auditory-visual temporal integration measured by shifts in perceived temporal location. AB - The perceived time of occurrence of a visual stimulus may be shifted towards the onset of an auditory stimulus occurring a short time later. The effect has been attributed to auditory-visual temporal integration although an unknown portion of the shift may be explained by the different processing times of visual and auditory stimuli. Here, perceived onset time is measured in a novel way that separates and compares the magnitude of these effects. Participants observed either a sequence consisting of a visual stimulus followed by an auditory stimulus and then another visual stimulus or the reverse. The temporal location of the second stimulus was varied systematically between the onset of the first and third stimuli, which were separated by a fixed duration. Two timescales were used: a short timescale that allowed for auditory-visual temporal integration to occur, and a long timescale that did not. Psychometric curves were fitted for both timescales, to the percentage the first interval was perceived is shortest, as a function of first interval duration. For the long timescale condition the point of subjective equality (PSE) of the two interval lengths was consistent with the different processing latencies. When visual and auditory stimuli occurred within 125 ms significant additional shifting of the PSE occurred. These results indicate that temporal integration shifts the perceived timing of a visual stimulus by an amount much larger than can be explained differential processing latencies. PMID- 17428608 TI - Mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequence of Spirocerca lupi (Nematoda, Spirurida): avenues for potential implications. AB - Canine spirocercosis is a life-threatening parasitosis caused by Spirocerca lupi (Nematoda, Spirurida) that is presently emerging in several countries. This study characterised an informative region within the mitochondrial (mtDNA) gene encoding for the cytochrome c oxidase subunit 1 (cox1) of S. lupi by Polymerase Chain Reaction (PCR)-coupled sequencing. Specimens from five different countries in Europe, Asia and Africa were examined and two different sequence variants of cox1 (i.e. haplotypes) were determined, displaying nucleotidic variation at 6 of 689 positions. All of these positions were invariable among all the parasite individuals from Europe (haplotype 1) and among the African and Asian individuals (haplotype 2), but differed between Europe and Asia/Africa. The S. lupi cox1 sequences were consistent with those of other common Spirurida previously reported at both nucleotidic and phylogenetic levels. This study provides molecular information essential for identification of the nematode, irrespective of its life cycle stage. Crucial implications for the specific molecular diagnosis of clinical spirocercosis and investigation of the evolution, population genetics, ecology and epidemiology of S. lupi are discussed. PMID- 17428609 TI - Flavonoids and their oxidation products protect efficiently albumin-bound linoleic acid in a model of plasma oxidation. AB - Although LDL esterified polyunsaturated fatty acids (PUFA) contribute largely to the pool of oxidizable lipids in plasma, they coexist with a non-negligible content of free PUFA. In some pathological conditions, the free PUFA/albumin ratio becomes abnormally elevated. Modeling was performed in a system constituted of linoleic acid bound to human serum albumin (HSA) in which oxidation was initiated by hydrophilic AAPH. Inhibition of lipid peroxidation was evaluated for various flavonoids. The accumulations of hydroperoxyoctadecadienoic acids (HPODE), hydroxyoctadecadienoic acids (HODE) and ketooctadecadienoic acids (KODE) were similarly inhibited: isoquercitrin>quercetin>catechin=isorhamnetin>>kaempferol>quercetin-4'-beta-D glucoside=quercetin-3,4'-di-beta-D-glucoside. Surprisingly, quercetin and isorhamnetin afforded a protection to linoleic acid long after their consumption. Elucidation by mass spectrometry and NMR of the quercetin oxidation products and assessment of their antioxidant capacity pointed out that 3,4-dihydroxybenzoic acid and 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxybenzofuran-3(2H)-one are major contributors to the apparent quercetin antioxidant capacity. PMID- 17428610 TI - A choice behavior for morphine reveals experience-dependent drug preference and underlying neural substrates in developing larval zebrafish. AB - Transparent larval zebrafish offer the opportunity to unravel genetic and neuronal networks underlying behavior in a developing system. In this study, we developed a choice chamber paradigm to measure reward-associated behavior in larval zebrafish. In the chamber where larval zebrafish have a choice of spending their time in either a water- or morphine-containing compartment, larvae that have previously experienced morphine spend significantly more time in the compartment containing morphine. This behavior can be attentuated by pre treatment with antagonists of the opioid receptor or the dopamine receptor, and furthermore, is impaired in the too few mutant, which has a genetic deficiency in the production of specific groups of dopaminergic and serotonergic neurons in the ventral forebrain. These results uncover a choice behavior for an addictive substance in larval zebrafish that is mediated through central opioid and monoaminergic neurotransmitter systems. PMID- 17428611 TI - Sex differences in the effect of finasteride on acute ethanol withdrawal severity in C57BL/6J and DBA/2J mice. AB - The neurosteroid allopregnanolone (ALLO) is a potent positive modulator of GABAA receptors that can modulate ethanol (EtOH) withdrawal. The 5alpha-reductase inhibitor finasteride can block the formation of ALLO and other GABAergic neurosteroids and also reduce certain effects of EtOH. Treatment with finasteride during chronic EtOH exposure decreased EtOH withdrawal severity and blood EtOH concentrations (BECs), suggesting an additional effect of finasteride on EtOH pharmacokinetics. Thus, the purpose of the present study was to determine the effect of finasteride on acute EtOH withdrawal severity, to minimize the effect of finasteride on EtOH metabolism. Male and female C57BL/6J and DBA/2J mice received a pretreatment of finasteride (50 mg/kg i.p.) or vehicle 24 h prior to an injection of EtOH (4 g/kg i.p.) or saline. Handling-induced convulsions (HICs) were scored at baseline, and then over a 24 h period after EtOH or saline injection. In another experiment, plasma estradiol and corticosterone levels were assessed at selected time points (0, 2, 8, and 24 h). In a final study, retro orbital blood samples were collected at 30, 60, 120, and 240 min post-EtOH administration to access finasteride's effects on EtOH clearance parameters. Pretreatment with finasteride increased acute EtOH withdrawal severity in female C57BL/6J and DBA/2J mice but decreased withdrawal severity in male mice of both strains. Finasteride did not alter BECs, EtOH clearance, estradiol, or corticosterone concentrations in a manner that appeared to contribute to the sex difference in finasteride's effect on acute EtOH withdrawal severity. These findings suggest that male and female C57BL/6J and DBA/2J mice differ in their sensitivity to changes in ALLO or other GABAergic neurosteroid levels during acute EtOH withdrawal. Sex differences in the modulation of GABAergic 5alpha reduced steroids may be an important consideration in understanding and developing therapeutic interventions in alcoholics. PMID- 17428612 TI - Alkmaion's discovery that brain creates mind: a revolution in human knowledge comparable to that of Copernicus and of Darwin. AB - Without special examination the brain offers no clue that it is the organ of the mind. From the dawn of time man thus either ignored the problem as to the source of thought, or attributed it to a variety of anatomical structures, usually the heart. The brain held no place in such intuitions, and in most languages it is analogized to bone marrow. Furthermore, nothing in early medical systems claimed any intellectual capacity for the brain; and the Egyptians, so fastidious in care for their afterlife, heedlessly discarded the brain in funerary practice. It was thus a unique event in world history when Alkmaion of Kroton (Alcmaeon, ca. 500 bc), based on anatomical evidence, proposed that the brain was essential for perception. Although no writings of Alkmaion survived, it was probably via a fortuitous linkage that his idea of the mental primacy of the brain was transmitted to, and preserved within, the teachings of the Hippocratic school. Nothing, of course, was secure as to mechanism, two millennia unfolding until the search for mind passed from the ventricles to the cerebral cortex. Nonetheless, Alkmaion was the beginning, and the ensuing understanding that he initiated is still transforming humanity's perception of the natural world, and their place within it. PMID- 17428613 TI - Comparison of in vivo and in vitro gene expression profiles in subventricular zone neural progenitor cells from the adult mouse after middle cerebral artery occlusion. AB - Stroke stimulates neurogenesis in the adult rodent brain. The molecules that mediate stroke-induced neurogenesis are not definitely known. Using microarrays containing approximately 400 known genes associated with stem cell and angiogenesis, we compared transcriptional profiles of subventricular zone (SVZ) tissue with cultured neural progenitor cells isolated from the SVZ 7 days after ischemic stroke in the adult mouse. In SVZ tissue, we found that stroke upregulated 58 genes which are involved in multiple signaling pathways during embryonic development, suggesting that stroke recaptures embryonic molecular signals. In neural progenitor cells cultured in growth medium, 23 gene expressions were increased after stroke and 8 of 23 genes overlapped with upregulated genes in stroke SVZ tissue. Expression alterations of selected genes were confirmed by real-time RT-PCR and immunohistochemistry. These in vivo and in vitro data provide new insight into the genetic program of adult SVZ neural progenitor cells after stroke and demonstrate gene expression differences between SVZ tissue and cultured SVZ cells. PMID- 17428614 TI - Prostaglandin transporter expression in mouse brain during development and in response to hypoxia. AB - Prostaglandins (PGs) are bioactive lipid mediators released following brain hypoxic-ischemic injury. Clearance and re-uptake of these prostaglandins occur via a transmembrane prostaglandin transporter (PGT), which exchanges PG for lactate. We used Western blot analyses to examine the PGT developmental profile and its regional distribution as well as changes in transporter expression during chronic hypoxia in the neonatal mouse brain. Microsomal preparations from four brain regions (cortex, hippocampus, cerebellum and brainstem/diencephalon) showed gradual increases in prostaglandin transporter expression in all brain regions examined from postnatal day 1 till day 30. There was a significant regional heterogeneity in the prostaglandin transporter expression with highest expression in the cortex, followed by cerebellum and hippocampus, and least expressed in the brainstem/diencephalon. To further delineate the pattern of prostaglandin transporter expression, separate astrocytic and neuronal microsomal preparations were also examined. In contrast to neurons, which had a robust expression of prostaglandin transporters, astrocytes had very little PGT expression under basal conditions. In response to chronic hypoxia, there was a significant decline in PGT expression in vivo and in neurons in vitro, whereas cultured astrocytes increased their PGT expression. This is the first report on PGT expression in the CNS and our studies suggest that PGTs have 1) a widespread distribution in the CNS; 2) a gradual increase and a differential expression in various regions during brain development; and 3) striking contrast in expression between glia and neurons, especially in response to hypoxia. Since PGTs play a role as prostaglandin-lactate exchangers, we hypothesize that PGTs are important in the CNS during stress such as hypoxia. PMID- 17428615 TI - Smoke and mirrors: magnified beliefs that cigarette smoking suppresses weight. AB - Research suggests that for some smokers, weight concerns interfere with smoking cessation. Studies with individuals with eating disorders and weight concerns have indicated that weight-concerned individuals place undue faith in the effectiveness of certain weight control strategies; i.e., adopt a brand of magical thinking pertaining to food rules and dieting behaviors. The current study investigated whether weight-concerned smokers endorsed exaggerated beliefs in the ability of smoking to suppress body weight. Participants were 385 individuals undergoing treatment for smoking cessation. Prior to treatment, participants completed the Smoking Consequences Questionnaire-Adult (SCQ-A), the Dieting and Bingeing Severity Scale, and the Perceived Risks and Benefits Questionnaire (PBRQ). Results indicated that heightened beliefs in the effectiveness of smoking to control weight were related to eating and weight concerns; specifically, strong associations were observed between SCQ-A Weight Control scores and fear of weight gain, loss of control over eating, and body dissatisfaction. Although SCQ-A Weight Control scores were related to (self reported) weight gain during a previous quit attempt, scores did not predict actual weight gain over the course of the cessation trial. Reported weight gain at previous attempts was also unrelated to actual weight gain over the current trial. These findings indicate that eating and weight-concerned smokers may benefit from psychoeducation concerning the relatively modest and temporary ability of nicotine to suppress weight. PMID- 17428616 TI - Frequency of heavy drinking and perceived peer alcohol involvement: comparison of influence and selection mechanisms from a developmental perspective. AB - The present study investigated social influence and selection explanations for the association between frequency of heavy drinking and perceived peer alcohol involvement in emerging and early adulthood. Participants were 489 young adults recruited from a university setting who were taking part in an 11-year longitudinal study, which includes 6 waves of data. Piecewise latent growth curve analyses indicated that patterns of change from ages 18 to 30 for both frequency of heavy drinking and perceived peer alcohol involvement are best represented by two distinct developmental periods (i.e., college and post-college years). Several models were compared to identify a framework that yielded the best estimates of influence and selection effects. Evidence suggests that selection mechanisms may best account for the relation between frequency of heavy drinking and perceived peer alcohol involvement especially during the post-college years. PMID- 17428617 TI - Motivational enhancement therapy for high-risk adolescent smokers. AB - The majority of regular adult smokers begin smoking in adolescence and there is a clear need for youth-targeted smoking cessation interventions. The present randomized, controlled trial tested the effectiveness of motivational enhancement therapy (MET) to reduce smoking among 81 adjudicated adolescents. Participants received either MET or an education control. Smoking abstinence, quantity, and frequency were assessed at 1 and 6 months post treatment. Results suggest that although between-group differences on outcome measures were not significant at follow-up, smoking behavior decreased in both groups with approximately 10% achieving 1-month smoking abstinence at 6-month follow-up. Furthermore, participant response to MET varied by level of alcohol use and impulsivity such that participants with lower levels of alcohol use and impulsivity had significantly greater response to MET. In contrast, participants who endorsed higher rates of alcohol use and impulsivity responded better to the control than the MET condition. Results suggest that MET may be an effective intervention for some adolescent smokers but may be contraindicated for adolescents who have concomitant problems with alcohol use or impulsivity. PMID- 17428618 TI - beta(3)-Adrenoceptor antagonism improves clinical outcomes of chronic heart failure. AB - Chronic heart failure (CHF) is a leading cause of morbidity and mortality throughout the world. The pathogenesis of CHF is complex but beta(1) adrenoceptors are critical in the process, because beta(1)-adrenoceptor blockers have been shown to significantly reduce the mortality and hospitalization rates in patients with CHF. Recent animal and human studies have demonstrated that there is an over expression of beta(3)-adrenoceptors in the failing heart, and stimulation of these receptors leads to further depression in ventricular function. We hypothesize that beta(3)-adrenoceptors and their over activities are one of the critical mechanisms of CHF, and addition to conventional heart failure therapies, beta(3)-adrenoceptor antagonism would further improve cardiac function and clinical outcomes. PMID- 17428619 TI - Huntington's disease and HDACi: would sulpiride and valproate be of therapeutic value? PMID- 17428620 TI - Body mass, polymorphisms in obesity-related genes, and the risk of developing breast cancer among women with benign breast disease. AB - BACKGROUND: A cohort study was conducted among post-menopausal women to determine whether genetic polymorphisms in selected obesity-related genes (PPARG, LPL, LEPR, PON1, PON2, TNF-alpha) were associated with the progression of benign breast disease (BBD) to breast cancer and whether the selected polymorphisms modified the association between body mass and breast cancer among women with BBD. METHODS: Among participants in an ongoing cohort study, 994 Caucasian post menopausal women had a breast biopsy for BBD. Of these women, 61 subsequently developed breast cancer. A short questionnaire was administered at baseline in 1989. Genotypes were determined using DNA extracted from blood collected in 1989. RESULTS: In this cohort, body mass index (BMI) was positively associated with the risk of developing breast cancer. In contrast, polymorphisms in PON1 (Gln192Arg) and LEPR (IVS2+6920) were associated with a decreased risk of developing invasive breast cancer. No statistically significant associations were observed for polymorphisms in PPARG, PON2, LPL, or TNF and breast cancer risk or for interactions between the polymorphisms and BMI and breast cancer risk. CONCLUSIONS: The findings suggest that specific polymorphisms in the PON1 and LEPR genes may play a role in progression of BBD to breast cancer among post menopausal Caucasian women. PMID- 17428621 TI - Social learning in horses from a novel perspective. PMID- 17428622 TI - Indicators of use of methamphetamine and other substances among men who have sex with men, San Francisco, 2003-2006. AB - BACKGROUND: Substance use has been associated with high-risk sexual behavior among men who have sex with men (MSM) both in the U.S. and around the world. Recent efforts by local organizations in San Francisco have specifically targeted methamphetamine use in this population. METHODS: We tracked methamphetamine and other substance use among men who have sex with men (MSM) in San Francisco from 2003 to 2006 using an indicator available in community outreach surveys of a prevention education program targeting MSM (n=4602). RESULTS: Overall, use of diverse substances tended to decrease from 2003 to 2006, many significantly so. Reported use of methamphetamine significantly decreased among HIV-negative MSM. However, methamphetamine and alcohol use during sex was associated with unprotected potentially HIV serodiscordant sex. CONCLUSION: Intensified prevention efforts to reduce methamphetamine use in San Francisco may be having some impact; however, strong associations of substance use and high-risk sex persist. PMID- 17428623 TI - Decreased susceptibility to doxycycline associated with a 16S rRNA gene mutation in Brachyspira hyodysenteriae. AB - The objective of this study was to assess whether nucleotide substitutions in the 16S rDNA sequence of selected Brachyspira hyodysenteriae isolates could explain differences in doxycycline minimal inhibitory concentrations (MICs). The main part of the 16S rRNA gene was sequenced and compared for 19 isolates with different doxycycline MICs. A mutation in the 16S rRNA gene at the position corresponding to 1058 in Escherichia coli has been shown to cause tetracycline resistance in other bacteria. In the B. hyodysenteriae sequences a G1058C mutation was found for all isolates with increased doxycycline MICs whereas all susceptible isolates had the wild type sequence. PMID- 17428624 TI - Protein tyrosine nitration and poly(ADP-ribose) polymerase activation in N-methyl N-nitro-N-nitrosoguanidine-treated thymocytes: implication for cytotoxicity. AB - 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG) is a DNA alkylating agent. DNA alkylation by MNNG is known to trigger accelerated poly(ADP-ribose) metabolism. Various nitroso compounds release nitric oxide (NO). Therefore, we set out to investigate whether MNNG functions as NO donor and whether MNNG-derived NO or secondary NO metabolites such as peroxynitrite contribute to MNNG-induced cytotoxicity. MNNG in aqueous solutions resulted in time- and concentration dependent NO release and nitrite/nitrate formation. Moreover, various proteins in MNNG-treated thymocytes were found to be nitrated, indicating that MNNG-derived NO may combine with cellular superoxide to form peroxynitrite, a nitrating agent. MNNG also caused DNA breakage and increased poly(ADP-ribose) polymerase activity and cytotoxicity in thymocytes. MNNG-induced DNA damage (measured by the comet assay) and thymocyte death (measured by propidium iodide uptake) was prevented by the PARP inhibitor PJ-34 and by glutathione (GSH) or N-acetylcysteine (NAC). The cytoprotection provided by PJ-34 against necrotic parameters was paralleled by increased outputs in apoptotic parameters (caspase activity, DNA laddering) indicating that PARP activation diverts apoptotic death toward necrosis. As MNNG induced cytotoxicity showed many similarities to peroxynitrite-induced cell death, we tested whether peroxynitrite was responsible for at least part of the cytotoxicity induced by MNNG. Cell-permeable enzymic antioxidants (superoxide dismutase and catalase), the NO scavenger cPTIO or the peroxynitrite decomposition catalyst FP15 failed to inhibit MNNG-induced DNA breakage and cytotoxicity. In conclusion, MNNG induces tyrosine nitration in thymocytes. Furthermore, MNNG damages DNA by a radical mechanism that does not involve NO or peroxynitrite. PMID- 17428625 TI - Estrogenic effects of mixtures of phyto- and synthetic chemicals on uterine growth of prepubertal rats. AB - Through the diet humans are exposed to many weak estrogenic phytochemicals (PCs) and synthetic chemicals (SCs), but most experimental studies used individual compounds rather than mixtures. Estrogenic effects were determined in the rat juvenile uterotrophic assay using a predefined phytochemical mixture (PCmix) containing coumestrol, genistein, naringenin, (+,-)catechin, (-,-)epicatechin and quercetin, and a predefined synthetic chemical mixture (SCmix) containing nonyl-, and octylphenol, beta-hexachlorocyclohexane, methoxychlor, bisphenol A and dibutylphthalate. The mixture composition was based on human dietary uptake and actual ratios in serum. 17beta-Estradiol and genistein were also tested individually. It was found that combinations of phytoestrogens and exogenous 17beta-estradiol act additive. In contrast SCmix, inactive by itself even at high dose levels relative to human exposure, caused no synergistic or antagonistic uterotrophic effect with E(2) and/or the PCmix. Based on ED(05) and ED(01) values of the PCmix the margin of exposure in regular human diet for a uterotrophic effect is estimated many orders of magnitude. However, food supplements with phytochemicals might bring individual exposure around ED(05) and ED(01) values of the PCmix. Based on the results of our study the contribution of SCs to total estrogenicity in human diet can probably be neglected. PMID- 17428626 TI - Endocrine and ovarian response after a 2-day controlled suckling and eCG treatment in lactating rabbit does. AB - Synchronization methods are used to obtain higher fertility when artificial insemination (AI) is applied to lactating rabbit does. The most common methods are eCG administration or temporary doe-litter separation. Nevertheless, drawbacks have been reported, such as negative side effects of hormonal treatment in the doe and low litter growth due to absence of suckling, respectively. Recently, improved reproductive performance (without visible consequences on young rabbit growth), has been obtained by applying a 2-day controlled nursing method before AI, by allowing for a 10 min nursing of the litter 24 h of separation. The present study was undertaken to examine the pituitary (PRL, LH, FSH) and the ovarian response (follicle size and number) to those methods. A total of 442 lactating does inseminated on day 11 post-partum were distributed in three experimental groups: 2CN (closing of nest box on day 9, controlled nursing on days 10 and 11), eCG (20 IU administered on day 9 post-partum) and CONTROL (untreated). Blood samples were obtained from 10 does per group at 48, 24 and 0 h before AI, and 1h after AI. Both 2CN and eCG treatments similarly improved sexual receptivity (76.3, 77.5 and 58.2%, respectively; P<0.001) and fertility (63.1, 64.1 and 48.4%, respectively; P<0.05) in lactating does, compared to the CONTROL group. Similar plasma FSH levels in all groups of does and sampling times were observed. Due to the absence of suckling, plasma concentration of PRL on day 10 post-partum in the 2CN group was lower than in the CONTROL group (P<0.05); this endocrine change in PRL levels could explain the better reproductive performances obtained with 2CN treatment. At 1h after exogenous administration of GnRH (at the moment of AI) a high LH response was observed in all groups (P<0.001). Ovaries from 20 rabbits treated in the same way but uninseminated (2CN, n=10; eCG, n=5; CONTROL, n=5 does) were obtained on day 11 post-partum in order to check the morphometric status (weight, width and height) and to make histological and immunohistochemical studies to detect growth hormone receptor (GH-R). As a result, synchronization methods did not show any significant difference in relation to the CONTROL group. However, a small increase in the number of primary follicles was evidenced in the 2CN group with respect to the eCG group, similarly to the CONTROL group (23.0+/-3.7, 9.4+/-4.9 and 14.8+/-4.92 primary follicles, respectively; P=0.1). GH-R immunostaining-presence was more evident in the 2CN and the eCG groups, including primordial follicles and oocytes themselves. Thus, there could have been some direct effects of GH on follicular development, as described in other species. Some ovarian parameters described open new ways to study intra-ovarian mechanism of follicular development in the post-partum period of rabbit does. PMID- 17428627 TI - Solid lipid nanoparticles loaded with insulin by sodium cholate phosphatidylcholine-based mixed micelles: preparation and characterization. AB - Solid lipid nanoparticles (SLNs) loaded with insulin-mixed micelles (Ins-MMs) were prepared by a novel reverse micelle-double emulsion method, in which sodium cholate (SC) and soybean phosphatidylcholine (SPC) were employed to improve the liposolubility of insulin, and the mixture of stearic acid and palmitic acid were employed to prepare insulin loaded solid lipid nanoparticles (Ins-MM-SLNs). Some of the formulation parameters were optimized to obtain high quality nanoparticles. The particle size and zeta potential measured by photon correlation spectroscopy (PCS) were 114.7+/-4.68 nm and -51.36+/-2.04 mV, respectively. Nanospheres observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM) showed extremely spherical shape. The entrapment efficiency (EE%) and drug loading capacity (DL%) determined with high performance liquid chromatogram (HPLC) by modified ultracentrifuge method were 97.78+/-0.37% and 18.92+/-0.07%, respectively. Differential scanning calorimetry (DSC) of Ins-MM-SLNs indicated no tendency of recrystallisation. The core-shell drug loading pattern of the SLNs was confirmed by fluorescence spectra and polyacrylamide gel electrophoresis (PAGE) which also proved the integrity of insulin after being incorporated into lipid carrier. The drug release behavior was studied by in situ and externally sink method and the release pattern of drug was found to follow Weibull and Higuchi equations. Results of stability evaluation showed a relatively long-term stability after storage at 4 degrees C for 6 months. In conclusion, SLNs with small particle size, excellent physical stability, high entrapment efficiency, good loading capacity for protein drug can be produced by this novel reverse micelle-double emulsion method in present study. PMID- 17428628 TI - Molecular interactions between dimethoxycurcumin and Pamam dendrimer carriers. AB - Dimethoxycurcumin, a lipophilic analog of curcumin found as a major pigment in the Indian species turmeric (Curcuma longa Linn.), is known to possess significant activity against various cancer cell lines, but its use as an anticancer drug is hindered by its poor water solubility. The conjugation of dimethoxycurcumin to water-soluble PAMAM dendrimers (generations 3.5 and 4) is demonstrated. The maximum drug-dendrimer incorporation efficiency is 4.3 and 5.0 molar for G3.5 and G4, respectively. The FTIR-ATR investigation of the neat compounds and the drug-dendrimer systems indicate that dimethoxycurcumin is in the enolic form, while its interaction with the integer generation dendrimer involves the major conformational change of the terminal ethylene amine groups. PMID- 17428629 TI - Anti-angiogenic activity of the methanol extract and its fractions of Ulmus davidiana var. japonica. AB - This study aimed to elucidate anti-angiogenic activity of Ulmus davidiana var. japonica that has been widely used in folk medicine. The methanol extract (UDE) of Ulmus davidiana var. japonica concentration-dependently displayed a strong inhibition in the chick chorioallantoic membrane (CAM) angiogenesis. The n butanol fraction of UDE and subsequent 30% MeOH subfraction were identified to be most responsible for the anti-angiogenic activity. PMID- 17428630 TI - Antimicrobial activity of the methanolic extract, fractions and four flavonoids from the twigs of Dorstenia angusticornis Engl. (Moraceae). AB - The methanolic extract (DAT), fractions (FRS) and four flavonoids, namely Gancaonin Q (1), Stipulin (2), Angusticornin B (3) and Bartericin A (4), isolated from the twigs of Dorstenia angusticornis (Moraceae), were tested for their in vitro antimicrobial activity. A total of 22 microbial cultures belonging to three Candida species, 6 Gram-positive and 13 Gram-negative bacterial species were used in this study. The inhibition zones (IZ) of the test samples against the pathogens were determined by the Agar Hole Diffusion test while the Liquid dilution method was used to determine their minimal inhibition concentrations (MIC) and their minimal microbicidal concentrations (MMC). Results indicate that DAT, compounds 3 and 4 inhibited the growth of all test pathogens. DAT, FRS 3-6, compounds 3 and 4 were both antibacterial and anticandidal. A single-dose oral toxicity performed in accordance with the OPPTS 870.1100 and OECD 401guideline showed that DAT was not toxic. Our findings provide a possible basis for the potential use of twigs from Dorstenia angusticornis in the treatment of infectious diseases. PMID- 17428631 TI - Sweat testing in addicts under methadone treatment: an Italian experience. AB - In the last years the interest in monitoring drug exposure with human sweat as alternative biological fluid, is increasing. Sweat collection is convenient, less invasive and difficult to adulterate compared to traditional specimens. The objective of this study was to determine the excretion profile of methadone and other drugs into human sweat. Pharmscope sweat patches (Medical Europe Diagnostic, Madrid, Spain) were used on heroin abusers under methadone treatment. Sweat patches were applied to 10 heroin addicts and 3 drug free volunteers admitted into the study. Sweat patches were worn for about 1 week; urine, saliva and hair samples were collected at the time of the removal of patches. After the extraction, sweat eluates were directly analyzed by GC/MS for the presence of nicotine, cotinine, caffeine, methadone, EDDP and cocaine. The extracts were subsequently derivatized to detect benzoylecgonine, ecgonine methyl ester, morphine, codeine and 6-acetylmorphine. No false positive results were obtained on the drug free samples. All the patches showed positive results for methadone. Cocaine was detected in two cases. Mainly the parent drug was identified rather than the metabolites. The results obtained show the usefulness of sweat as complementary specimen to saliva and urine providing a longer detection window. Moreover, sweat testing offers the advantage of being a non-invasive means of obtaining information about drug exposure. PMID- 17428632 TI - Intelligent bioprocessing for haemotopoietic cell cultures using monitoring and design of experiments. AB - The need for successful ex-vivo expansion and directed differentiation of haematopoietic stem cells (HSCs) for therapeutic applications has increased over the past decade. Haematopoietic cell cultures are complex and full characterisation of the process environment has yet to be achieved. The complexity and transient nature of HSC cultures make the identification, maintenance and control of optimal operating conditions challenging. Application of real-time, on-line monitoring techniques and process control strategies enhances the ability to operate bioprocesses of desired reproducibility and high product quality. In this review, we discussed the methods by which in vitro culture information necessary for bioprocess control may be obtained, including process considerations, monitoring and analytical tools, and design of experiments (DOE). The successful application of these tools may result in time- and cost-effective cultures for directed differentiation and expansion of haematopoietic components intended for clinical use. PMID- 17428633 TI - Alterations of NR2B and PSD-95 expression after early-life epileptiform discharges in developing neurons. AB - As an extreme form of abnormally synchronized activity, epilepsy may modify patterns of organization in the nervous system. It is clear that enhanced glutamatergic excitatory synaptic transmission with alterations in the expression of ionotropic glutamate receptors is a mechanism critical for seizure susceptibility and excitotoxicity. However, the exact quomodo and the roles of regulated N-methyl-D-aspartate receptor (NMDAR) composition and expression of a major postsynaptic density (PSD) scaffolding molecule, PSD-95, are as yet unclear. To study protein expression changes after epileptiform discharges in cultured immature rat cortical neurons, we divided cells into three groups which were transiently exposed to regular Neurobasal/B27 (control group), physiological solution (PS group) and magnesium-free physiological solution (MGF group) at cultured day 6. Neurons at three different culture ages (DIV7, DIV12 and DIV17) were collected for immunoblotting analysis. We found a decrease in expression of NR2B NMDAR subunit and PSD-95 (P<0.05) shortly after insult (within 24 h), which may show that brief magnesium-free media treatment of primary cultured rat cortical neurons, an in vitro model of seizure brain injury, has a major influence on the expression of NR2B subunit and PSD-95. PMID- 17428634 TI - Conflict, collusion or collaboration in consultations about medically unexplained symptoms: the need for a curriculum of medical explanation. AB - OBJECTIVE: To identify the basis of the communication problems that characterise consultations about medically unexplained symptoms (MUS) and to identify implications for clinical education. METHOD: Recent research into the details of clinical communication about MUS was reviewed narratively and critically, and broader research literature was scrutinised from the perspective of a practitioner who wishes to provide patients with explanations for such symptoms. RESULTS: Consultations about MUS often involve contest between patients' authority, resting on their knowledge of their symptoms, and practitioners' authority, based on the normal findings of tests and investigations. The outcome of consultations can therefore depend on the strategies that each party uses to press their authority, rather than on clinical need. CONCLUSION: Contest is a product of patients and practitioners occupying separate conceptual 'ground'. Avoiding contest requires the practitioner to find common conceptual ground within which each party can understand and discuss the symptoms. Finding common ground by collusion with explanations that patients suggest can damage clinical relationships. Instead the practitioner needs to fashion explanation that is acceptable to both parties from available medical and lay material. PRACTICE IMPLICATIONS: Although practitioners commonly fashion such explanations, this aspect of their professional role seems not to be greatly valued amongst practitioners or in medical curricula. Clinical education programmes could include curricula in symptom explanation, drawing from research in medicine, psychology and anthropology. PMID- 17428635 TI - Gastric GIST: a single institutional retrospective experience with surgical treatment for primary disease. AB - AIMS: Because gastric GISTs show variable clinical behavior, we reviewed our experience with primary gastric GISTs after surgical treatment and imatinib mesylate treatment for advanced disease. METHODS: Between December 1995 and December 2005, 111 patients who underwent surgical treatment for primary gastric GISTs were enrolled in this study. Patients were grouped according to the risk assessment classification, and clinicopathological features, tumor recurrence and patient survival were assessed. RESULTS: One patient was included in the very low risk group, 35 in the low risk group, 31 in the intermediate risk group and 44 in the high-risk group. All patients with very low, low and intermediate risk GISTs and 70% of patients with high risk GISTs underwent R0 resection. While there was no recurrence or metastasis in patients with very low, low and intermediate risk GISTs, 23% of those with high risk GISTs showed a distant metastasis at diagnosis and 35% of these patients had a recurrence after R0 resection. The overall 5-year survival rate of the high risk patients was 77.1%. Nineteen patients received imatinib mesylate therapy due to an incomplete resection or recurrence; 7 with no measurable lesion at the CT scan by a local tumor control showed no tumor progression after imatinib mesylate therapy, however, 12 patients with measurable lesions showed variable clinical courses after treatment. The overall 5-year survival rate of 19 patients with imatinib mesylate treatment was 80.0%. CONCLUSIONS: The clinical outcome of the very low, low and intermediate risk gastric GISTs was excellent, while high risk gastric GISTs had a high rate of recurrence and therefore a less favorable outcome. A complete resection is the most important treatment for cure; however imatinib mesylate treatment may improve the clinical outcome of the patients with metastatic or recurrent gastric GISTs. PMID- 17428636 TI - Infection-induced viscerosensory signals from the gut enhance anxiety: implications for psychoneuroimmunology. AB - Infection and inflammation lead to changes in mood and cognition. Although the "classic" sickness behavior syndrome, involving fatigue, social withdrawal, and loss of appetites are most familiar, other emotional responses accompany immune activation, including anxiety. Recent studies have shown that gastrointestinal bacterial infections lead to enhanced anxiety-like behavior in mice. The bacteria induced signal is most likely carried by vagal sensory neurons, and occurs early on (within 6h) during the infection. These signals induce evidence of activation in brain regions that integrate viscerosensory information with mood, and potentiate activation in brain regions established as key players in fear and anxiety. The findings underline the importance of viscerosensory signals arising from the gastrointestinal tract in modulation of behaviors appropriate for coping with threats, and suggest that these signals may contribute to affective symptoms associated with gastrointestinal disorders. PMID- 17428637 TI - Computerized tomography based on DEI refraction information. AB - Conventional computerized tomography (CT) technique is based on the absorption contrast. In recent years, X-ray phase-contrast CT (PCCT) has been developing rapidly. It uses the phase information to reconstruct the object and provide high contrast and spatial resolution. Diffraction enhanced imaging (DEI) method is one of the three phase-sensitive X-ray imaging techniques. DEI method employs an analyzer crystal to extract the object's refraction information which can be used for CT. However, when DEI refraction CT is combined with the conventional CT algorithm, it should be satisfied that the refraction information of an arbitrary point in the object is invariable at every projection view. In this paper, the invariance condition of refraction information is analyzed and two feasible methods are provided for reconstruction. Using these two methods, two samples of weak absorption are reconstructed with the experimental data obtained at Beijing Synchrotron Radiation Facility (BSRF). PMID- 17428638 TI - Towards building a photo-realistic virtual human face for craniomaxillofacial diagnosis and treatment planning. AB - The aim of this investigation was to assess the feasibility of building a virtual human face digitally by superimposing a photo-realistic three-dimensional (3D) soft-tissue surface on bone in the correct relationship and evaluating the registration errors associated with this method. The 3D soft-tissue surface of the face was captured using a fast stereophotogrammetry method and the underlying bone was recorded using a 3D computed tomography (CT) scanner. Using the Procrustes registration method, the outer surface of the 3D CT scan and the photo realistic soft-tissue surfaces were merged into a single Virtual Reality Modelling Language (VRML) file and displayed using a standard VRML viewer. Quantitative measurements of registration errors were calculated in the reconstructed human head models using the signed closest point distance from the photo-realistic skin surface to the transformed CT skin surface. The registration errors between most parts of the aligned surfaces were within +/-1.5mm. The errors were relatively large around the eyebrows, eyelids and cheeks. Simultaneous recording of the face and skull may reduce this error. PMID- 17428639 TI - Tolerogenic dendritic cells and regulatory T cells: a two-way relationship. AB - At first glance, dendritic cells (DCs) and regulatory T cells (Tregs) do not have much in common. DCs are characterized by their unsurpassed T cell stimulatory capacity, whereas Tregs are marked by the ability to suppress proliferation of effector T cells. However, only mature/activated DCs stimulate T cell proliferation, whereas immature DCs induce Tregs. This provides a means by which peripheral tolerance is maintained: in the absence of inflammation and disease, DCs encounter apoptotic cells and "self" detritus in peripheral tissues. Thus, DCs constantly sample the peripheral environment and, accordingly, the presentation of "self" by these steady state DCs results in induction of suppressive Tregs. Vice versa, Tregs are able to affect DC development, preventing maturation and inducing IL-10, as well as immunosuppressive molecules of the B7-H family, in DCs. Therefore, these novel findings establish a mutual interaction between DCs and Tregs for the upkeep of immunosuppression: immature DCs induce Tregs and inversely Tregs prepare DCs to become immunosuppressive, thereby extending the immunosuppressive function of Tregs. The possible means of cellular interactions as well as the consequences for tolerance and immunity are discussed in this review. PMID- 17428640 TI - Infective endocarditis caused by USA300 methicillin-resistant Staphylococcus aureus (MRSA). AB - We report seven cases of infective endocarditis caused by USA300 methicillin resistant Staphylococcus aureus (MRSA) at an urban, tertiary care, academic institution. Five strains were community associated and two were healthcare associated. All patients were injection drug users. Staphylococcus aureus isolates were characterised as USA300-type MRSA using pulsed-field gel electrophoresis. Five cases were right-sided endocarditis and two cases were left sided. The mean length of in-hospital antimicrobial therapy was 23 days and the mean length of total antibiotic therapy was 55 days. Complications included heart failure resulting in valve replacement in one patient as well as death in that patient. As USA300 strains of MRSA continue to increase in prevalence, clinicians must be aware of the increasing spectrum of illness in considering management and prevention strategies. PMID- 17428641 TI - Development, evaluation and application of an isocratic high-performance liquid chromatography (HPLC) assay for the simultaneous determination of aciclovir and its metabolite 9-carboxymethoxymethylguanine in human serum and cerebrospinal fluid. AB - 9-Carboxymethoxymethylguanine (CMMG), the main metabolite of aciclovir (ACV), is a putative neurotoxin. Measurement of CMMG in body fluids may aid patient management. We describe the development, validation and application of a high performance liquid chromatography (HPLC) method for the simultaneous determination of ACV and CMMG in human serum and cerebrospinal fluid (CSF). Recovery was between 94% and 100% at all concentrations both from serum (range 0 20 mg/L) and CSF (0-5 mg/L). The intra-assay precision (coefficient of variation (CV)) was <2% and the inter-assay precision (CV) was <5%. The limits of detection and quantification were 0.1 and 0.25 mg/L, respectively, in both body fluids. Significant interference from endogenous material or from drugs in clinical samples was not seen. CMMG was detected in most of the 55 clinical samples containing ACV, but little correlation was found between the levels of the drug and its metabolite. PMID- 17428642 TI - The role of Transient Receptor Potential Vanilloid 1 (TRPV1) channels in pancreatitis. AB - Premature activation of digestive enzymes within the pancreas which leads to autodigestion of the gland is an early step in the pathogenesis of pancreatitis. Pancreatic injury is followed by other manifestations of inflammation including plasma extravasation, edema, and neutrophil infiltration which constitute the features of pancreatitis. Recent studies indicate that neural innervation of the pancreas may play an important role in the initiation and maintenance of the inflammatory response to injury. The pancreas is innervated by vagal, sympathetic and parasympathetic neurons, as well as sensory neurons. Activation of pancreatic primary sensory neurons causes the release of inflammatory neuropeptides both in the spinal cord to signal pain and in the pancreas itself where they produce plasma extravasation and neutrophil infiltration. Recent studies indicate that primary sensory neurons of the pancreas express transient receptor potential V1 (TRPV1) channels whose activation induces pancreatic inflammation. Moreover, blockade of these TRP channels significantly ameliorates experimental pancreatitis. This review describes our current understanding of the role of TRPV1 channels in pancreatitis and illustrates how this mechanism might be used to direct future treatments of pancreatic diseases. PMID- 17428643 TI - Kinetic studies on the interactions between glycolipid biosurfactant assembled monolayers and various classes of immunoglobulins using surface plasmon resonance. AB - Kinetic studies on the interactions between self-assembled monolayers of mannosylerythritol lipids (MELs), which are glycolipid biosurfactants abundantly produced by microorganisms, and various classes of immunoglobulins including human IgG, IgA, and IgM were performed using surface plasmon resonance (SPR). The effect of the MEL structure on the binding behavior of HIgG was examined. Assembled monolayers of MEL-A having two acetyl groups on the headgroup gave a high affinity (K(d)=1.7x10(-6)M) toward HIgG, while those of MEL-B or MEL-C having only one acetyl group at C-6' or C-4' position gave little affinity. Our kinetic analysis revealed that the binding manner of HIgG, HIgA (K(d)=2.4x10( 7)M), and HIgM (K(d)=2.2x10(-7)M) to the assembled monolayers of MEL-A is not the monovalent mode but the bivalent mode, and both the first and second rate association constants (k(a1), k(a2)) increase with an increase in the number of antibody binding sites, while those for dissociation (k(d1), k(d2)) changed little. Moreover, we succeeded in directly observing great amounts of HIgG, HIgA, and HIgM bound to MEL-A monolayers using atomic force microscopy (AFM). Finally, we found that MEL-A assembled monolayer binds toward various IgG derived from mouse, pig, rabbit, horse, goat, rat, and bovine as well as human IgG (HIgG), and the only exception was sheep IgG. These results clearly demonstrate that MEL-A assembled monolayers would be useful as noble affinity ligand system for various immunoglobulins. PMID- 17428644 TI - Logical properties of foundational relations in bio-ontologies. AB - OBJECTIVE: We compare the advantages of specifying the semantics of foundational relations in bio-medical terminology systems using different types of formal deductive systems: first-order logic (FOL) and description logics (DLs). METHOD: As our focus example, we use a terminology whose basic terms are supposed to designate proper parthood relations, subdivision relations, and surrounded-by relations. Each type of relation captures an important and distinct aspect of the spatial organization of anatomical structures: the general part-whole structure (proper parthood), the division of salient anatomical objects into discrete, tree like structures (subdivision-of), and the nesting of anatomical objects into containers (surrounded-by). We show that all three types of relations are strict partial orderings (i.e., asymmetric and transitive). Ontologies whose purpose is to specify the semantics of terms referring to these types of relations must include axioms strong enough to formally distinguish among them. We compare the extent to which axioms characterizing proper parthood, subdivision, and surrounded-by relations can be represented in first-order logic and various description logics. CONCLUSIONS: The development of bio-medical ontologies requires a rigorous formal analysis of foundational relations. Different kinds of formal tools may be used in this process. Ideally, an analysis in a highly expressive language, such as first-order logic, should be worked out in conjunction with analyses in less expressive but computationally tractable deductive systems such as description logics. PMID- 17428645 TI - The isolation and characterization of virulence factors of Cryptococcus spp. from saprophytic sources in the city of Ribeirao Preto, Sao Paulo, Brazil. AB - Yeasts of the Cryptococcus genus are distributed in nature associated to animal and vegetal organic residues. Occasionally, species other than C. neoformans may be responsible for infectious diseases in human and animals. This study aims to determine the occurrence of Cryptococcus species in the atmosphere and bird droppings in the city of Ribeirao Preto, Sao Paulo, Brazil, and to evaluate three virulence factors: capsule formation, growth at 37 degrees C and melanin production. We analyzed 86 environmental samples (54 droppings and 32 air). Of the 41 strains isolated, 15 were C. neoformans var. neoformans (12 droppings and 3 air), 15 C. albidus (12 droppings and 3 air), 9 C. laurentii (7 droppings and 2 air) and 2 C. uniguttulatus (from droppings). Capsules were produced by 93.3% of C. neoformans var. neoformans, 66.7% of C. albidus, 88.9% of C. laurentii and 50% (1/2) of C. uniguttulatus. All strains of C. neoformans, 20% of C. albidus and 44.4% of C. laurentii were able to grow at 37 degrees C. The melanin production on DOPA agar was verified in C. neoformans (93.3%), C. albidus (26.7%) and C. laurentii (66.7%). We concluded that different Cryptococcus species coexist in the same ecological niche and they are able to produce virulence factors. PMID- 17428646 TI - Solubilization and characterization of a cell wall-bound trehalase from ascospores of the fission yeast Schizosaccharomyces pombe. AB - The genome of the fission yeast Schizosaccharomyces pombe lacks sequence homologs to ath1 genes coding for acid trehalases in other yeasts or filamentous fungi. However, acid trehalase activity is present at the spore stage in the life cycle of the fission yeast. The enzyme responsible for this activity behaves as a surface enzyme covalently linked to the spore cell walls in both wild-type and ntp1 mutant strains devoid of neutral trehalase. Lytic treatment of particulated cell wall fractions allowed the solubilization of the enzyme into an active form. We have characterized this soluble enzyme and found that its kinetic parameters, optimum pH and temperature, thermal denaturation and salt responses are closely similar to other conventional acid trehalases. Hence, this rather unusual enzyme can be recognized as acid trehalase by its biochemical properties although it does not share genetic homology with other known acid trehalases. The potential role of such acid trehalase in the mobilization of trehalose is discussed. PMID- 17428647 TI - Use of AFLP for differentiation of Metschnikowia pulcherrima strains for postharvest disease biological control. AB - Metschnikowia pulcherrima occurs naturally on fruits, buds and floral parts of apple trees. Some strains are effective as biocontrol agents against postharvest decay of apples and other fruits. The usefulness of the amplified fragment length polymorphism (AFLP) technique was evaluated for the genetic analysis of 26 strains of M. pulcherrima, isolated from different sources in different geographical regions. With six AFLP primer pairs, 729 polymorphic bands were scored. The technique showed a high discriminatory power. Genetic relationships between strains were also estimated using AFLP. All the isolates from the carposphere of apple, previously tested as biocontrol agents, were grouped in a single cluster with a high bootstrap value (97), indicating robustness and reproducibility. AFLP patterns could clearly distinguish the different strains and research is in progress to use some putative specific bands for single tag sequence (STS) conversion to develop isolate-specific markers. PMID- 17428648 TI - NKT cells: T lymphocytes with innate effector functions. AB - Natural killer T (NKT) cells are innate-like T lymphocytes that recognize glycolipid antigens in the context of the MHC class I-related glycoprotein CD1d. Recent studies have identified multiple ways in which NKT cells can become activated during microbial infection. Mechanisms of CD1d-restricted antigen presentation are being unraveled, and a surprising connection has been made to proteins that control lipid metabolism and atherosclerosis. It appears that several microorganisms have developed strategies to interfere with the CD1d antigen-presentation pathway. New studies have also provided important insight into the mechanisms that control effector cell differentiation of NKT cells and have revealed specialized functions of distinct NKT cell subsets. Finally, there is continued enthusiasm for the development of NKT cell-based therapies of human diseases. PMID- 17428649 TI - Two distinct clinical types of interstitial lung disease associated with polymyositis-dermatomyositis. AB - Most patients with interstitial lung disease (ILD) associated with collagen vascular diseases (CVD) have a chronic indolent course with a relatively favorable prognosis; however, acute progression has been reported in some polymyositis-dermatomyositis patients. This study evaluated the prevalence, clinical features, and outcome relative to the presentation type of ILD in polymyositis-dermatomyositis (PM-DM). Ninety-nine patients with newly diagnosed polymyositis-dermatomyositis seen at the Asan Medical Center in Korea between January 1990 and December 2004 were enrolled. The clinical, radiological, and pathological findings were retrospectively reviewed. ILD were divided into acute (dyspnea within 1 month before diagnosis) or chronic types. ILD was found on chest radiographs in 33 patients (33.3%), and 11 (33.3%) of these were considered acute. The acute group presented with more severe respiratory symptoms, hypoxemia, and poorer lung function. Patients with an acute presentation had ground glass opacity and consolidation on high-resolution computed tomography (HRCT), in contrast to reticulation and honeycombing in the chronic type. Surgical lung biopsy of one acute-type patient revealed diffuse alveolar damage, whereas biopsies in the chronic type showed usual interstitial pneumonia (UIP) in four cases and nonspecific interstitial pneumonia (NSIP) in another four. Eight acute-type patients (72.7%) died of respiratory failure within 1-2 months despite steroid therapy. The 3-year mortality rate of the chronic-type patients (21.2%) was not statistically significantly different from that of the patients without ILD (10.2%). In polymyositis-dermatomyositis, the acute, severe form of ILD was not infrequent. PMID- 17428650 TI - Identification of individuals with alpha-1-antitrypsin deficiency by a targeted screening program. AB - BACKGROUND: Alpha-1-antitrypsin deficiency (AATD) is significantly underdiagnosed. The early detection of AATD would enable affected persons to make lifestyle changes such as quitting smoking. It was the aim of the study to determine whether the combination of an awareness program with the offer of a cost-free diagnostic test results in the identification of a significant number of individuals with severe AATD. METHODOLOGY: We combined a series of measures to promote awareness with the offer of a diagnostic test at no charge. Test blood was applied to a filter paper and sent to our laboratory. The level of AAT was measured by nephelometry, the presence of the S- or Z-allele was determined by PCR, and phenotyping was performed by isoelectric focusing. RESULTS: During 37 months 17688 testing kits were distributed and 2722 were sent back to our laboratory. We identified 335 patients with severe AATD including 16 individuals with rare genotypes. Prescreening by determining the AAT serum levels by the submitting physician increased the detection rate as compared to similar programs that screened unselected individuals. SUMMARY: These data show that the combination of an awareness program with the offer of free diagnostic testing results in the identification of a large number individuals with severe AATD. PMID- 17428651 TI - Mounier-Kuhn syndrome: a rare cause of severe bronchial dilatation with normal pulmonary function test: a case report. AB - Tracheobronchomegaly (TBM) (Mounier-Kuhn syndrome) is dilatation of the trachea and major bronchi because of atrophy or absence of elastic fibers and smooth muscle cells. We present a case of TBM with normal pulmonary function test (PFT). The patient was a 37-year-old man with increasing productive cough and without fever, wheezes, chest pain, weight loss or any respiratory disease. Chest helical computed tomography (CT) scan showed tracheomegaly with transversal diameters of the trachea of 44mm. CT scan showed collapse of the trachea. Few large diverticular out-pouching and openings in the trachea was seen in bronchoscopy. PFT results were normal. PFT in large airway disorders may be normal while abnormalities may indicate underlying small airway disorder. An underlying small airway disorders is responsible for abnormal reports in PFT of these patients. We may need to re-evaluate the role of PFT within follow-up of patients with large airway disorder. PMID- 17428652 TI - Comparative evaluation of life cycle assessment models for solid waste management. AB - This publication compares a selection of six different models developed in Europe and America by research organisations, industry associations and governmental institutions. The comparison of the models reveals the variations in the results and the differences in the conclusions of an LCA study done with these models. The models are compared by modelling a specific case - the waste management system of Dresden, Germany - with each model and an in-detail comparison of the life cycle inventory results. Moreover, a life cycle impact assessment shows if the LCA results of each model allows for comparable and consecutive conclusions, which do not contradict the conclusions derived from the other models' results. Furthermore, the influence of different level of detail in the life cycle inventory of the life cycle assessment is demonstrated. The model comparison revealed that the variations in the LCA results calculated by the models for the case show high variations and are not negligible. In some cases the high variations in results lead to contradictory conclusions concerning the environmental performance of the waste management processes. The static, linear modelling approach chosen by all models analysed is inappropriate for reflecting actual conditions. Moreover, it was found that although the models' approach to LCA is comparable on a general level, the level of detail implemented in the software tools is very different. PMID- 17428653 TI - LCA-IWM: a decision support tool for sustainability assessment of waste management systems. AB - The paper outlines the most significant result of the project 'The use of life cycle assessment tools for the development of integrated waste management strategies for cities and regions with rapid growing economies', which was the development of two decision-support tools: a municipal waste prognostic tool and a waste management system assessment tool. The article focuses on the assessment tool, which supports the adequate decision making in the planning of urban waste management systems by allowing the creation and comparison of different scenarios, considering three basic subsystems: (i) temporary storage; (ii) collection and transport and (iii) treatment, disposal and recycling. The design and analysis options, as well as the assumptions made for each subsystem, are shortly introduced, providing an overview of the applied methodologies and technologies. The sustainability assessment methodology used in the project to support the selection of the most adequate scenario is presented with a brief explanation of the procedures, criteria and indicators applied on the evaluation of each of the three sustainability pillars. PMID- 17428654 TI - p73: a chiaroscuro gene in cancer. AB - p73 is a member of the p53 family which is gaining increasing importance in the field of cancer. Its structural homology with p53 led to the assumption that it could act as a new tumour suppressor gene. Increasing knowledge of its function, however, has cast doubts on this role. A particularly interesting characteristic of p73 is that the cell contains different isoforms with distinct and sometimes opposite functions. Evidence in the last few years clearly indicates that p73 does share some activities with p53 but also that it has some distinct functions. This review focuses on p73's role in the development and progression of cancer, analysing the gene structure and regulation and discussing similarities with p53 and differences. Recent results obtained with specific detection methods on the levels and functions of the different isoforms in tumours are also discussed. PMID- 17428655 TI - DT56a (Femarelle): a natural selective estrogen receptor modulator (SERM). AB - A selective estrogen receptor modulator (SERM) is defined as a substance with dissimilar effects on different tissues: agonist in some and antagonists in others. The natural compound DT56a (Femarelle) was shown to activate estrogen receptors in human cultured female derived osteoblasts. It was also shown to relieve menopausal symptoms and to increase bone mineral density with no effect on sex steroid hormone levels and on the endometrial thickness. DT56a, similarly to estradiol-17beta (E2), stimulated the specific activity of creatine kinase (CK) in skeletal and vascular tissues of female rats, as a marker of estrogen receptor (ER) activation. However, in the uterus, CK was activated only by E2 but not by DT56a. In order to prove that DT56a is a SERM, we examined the mutual interaction between DT56a and E2, at supra physiological doses, in different tissues in both intact and ovariectomized female rats, as well as in human cultured vascular and bone cells. Administration of DT56a or E2 stimulated CK in all tissues tested, but when given simultaneously, in intact immature female rats, DT56a completely abolished E2 stimulation of CK in all organs except in the diaphyseal bone where the inhibition was partial. In ovariectomized female rats, DT56a abolished E2's stimulation of CK in diaphyseal bone, thymus, uterus and pituitary but caused a partial inhibition in aorta, left ventricle and epiphyseal cartilage. In human bone cells E2 stimulation of CK, of alkaline phosphatase (AP) activity and of DNA synthesis was completely abolished by DT56a in post menopausal cells and partially inhibited in pre-menopausal cells. In human vascular cells, inhibition of DNA synthesis by E2 was completely abolished by DT56a and E2-induced CK was partially inhibited by DT56a. The results support the finding that DT56a is a SERM; it stimulated different parameters similar to E2, but when given simultaneously, at supra physiological doses, inhibited these E2's effects. Further investigations regarding intra and extra cellular mechanism of action of DT56a are currently performed. PMID- 17428656 TI - 3beta-Hydroxysteroid dehydrogenase/5-ene-4-ene isomerase mRNA expression in rat brain: effect of pseudopregnancy and traumatic brain injury. AB - Evidence that endogenous progesterone (PROG) is neuroprotective after traumatic brain injury (TBI) is supported by the findings that pseudopregnant female rats present less edema and achieve better functional recovery than do male rats. PROG in the nervous system may originate from steroidogenic glands or can be locally synthesized. 3beta-Hydroxysteroid dehydrogenase/5-ene-4-ene isomerase (3beta-HSD) is the key enzyme in the biosynthesis of PROG. In the present study, we investigated the effects of pseudopregnancy and TBI on brain 3beta-HSD mRNA expression and on PROG levels. Twenty-four hours after bilateral contusion of the medial prefrontal cortex of rats, 3beta-HSD mRNA expression was analyzed by in situ hybridization while PROG levels were measured by gas chromatography/mass spectrometry. Similar levels of 3beta-HSD mRNA expression were observed in males and pseudopregnant females in the non-injured groups. At this time point, there was a significant decrease in the 3beta-HSD mRNA expression in the contusion site within the frontal cortex in both males and pseudopregnant females. In all other regions analyzed, 3beta-HSD mRNA expression was not affected by TBI and there was no difference between males and pseudopregnant females. The high decrease in the expression of the 3beta-HSD mRNA in the lesion site 24 h after TBI suggests a possible decrease in locally synthesized PROG in lesion site without change in the other brain regions. This decrease has less impact in pseudopregnant females since they have high plasmatic and brain levels of PROG compared to males. PMID- 17428657 TI - Potent bradykinin B1 receptor antagonists: 4-substituted phenyl cyclohexanes. AB - Selective bradykinin (BK) B(1) receptor antagonists have been shown to be antinociceptive in animal models and could be novel therapeutic agents for the treatment of pain and inflammation. Elucidation of the structure-activity relationships of the biphenyl moiety of the lead compound 1 provided a potent new structural class of BK B(1) receptor antagonists. PMID- 17428658 TI - A tetra(L-lysine)-grafted poly(organophosphazene) for gene delivery. AB - In order to develop a new gene delivery vector, a novel cationic poly(organophosphazene) was synthesized by stepwise nucleophilic substitutions of poly(dichlorophosphazene) with a hydrophilic methoxy-poly(ethylene glycol) (MPEG) as a shielding group and a branched tetra(L-lysine), LysLys(LysEt)(2), as a cationic moiety. The cationic polymer has shown to form a polyplex by DNA condensation and very low in vitro cytotoxicity probably due to the shielding effect of MPEG, which provides a basis for improving the low gene transfection yield of cationic polyphosphazenes. PMID- 17428659 TI - Synthesis and structure-activity relationships of 4-hydroxy-4-phenylpiperidines as nociceptin receptor ligands: Part 1. AB - A series of 4-hydroxy-4-phenylpiperidines have been synthesized and bind to the nociceptin receptor with high affinity. The synthesis and structure-activity relationships at the N-1 and C-4 are described. PMID- 17428660 TI - Synthesis and structure-activity relationships of novel pyrimido[1,2-b]indazoles as potential anticancer agents against A-549 cell lines. AB - A series of novel pyrimido[1,2-b]indazoles 5, 7 have been prepared from 3 trifluoromethyl-5-phenyl-2,6-dicyano anilines 1 via novel indazole regioisomers 3 and 4 through a facile strategy. Specific examples were evaluated for anticancer activity in vitro and found to exhibit promising activity against A-549 cell lines and are more effective than Etoposide. QSAR models were developed and validated by cross-validation method. The results of the best QSAR model were further compared with the crystal structure of tubulin protein. The binding energies estimated were found to have a good correlation with the experimental inhibitory potencies. PMID- 17428661 TI - The thioesterase domain from the pimaricin and erythromycin biosynthetic pathways can catalyze hydrolysis of simple thioester substrates. AB - The recombinant polyketide synthase thioesterase domains from the pimaricin and 6 deoxyerythronolide B biosynthetic pathways catalyze hydrolysis of a number of simple N-acetylcysteamine thioester derivatives. This study demonstrates that thioesterases are not highly substrate selective in formation of the acyl-enzyme intermediate, in contrast to non-ribosomal peptide synthase thioesterase domains that show very high specificity for substrate loading. This observation has important implications for the engineering of biosynthetic pathways to produce polyketide products. PMID- 17428662 TI - Structure-based organic synthesis of unnatural aeruginosin hybrids as potent inhibitors of thrombin. AB - Based on X-ray crystallographic data of complexes of chlorodysinosin A with the enzyme thrombin, a series of analogs were synthesized varying the nature of the P(1), P(2), and P(3) pharmacophoric sites and the central octahydroindole carboxyamide core. In general, introduction of a hydrophobic substituent on the d leucine amide residue dramatically improved the inhibition of the enzyme. This is rationalized based on a better fit of the P(3) subunit in the hydrophobic S(3) enzyme site. Single digit nanomolar inhibition expressed as IC(50) was observed for several analogs. PMID- 17428663 TI - Glaziovianin A, a new isoflavone, from the leaves of Ateleia glazioviana and its cytotoxic activity against human cancer cells. AB - A new isoflavone, named glaziovianin A (1), was isolated from the leaves of Ateleia glazioviana (Legminosae) by means of a cytotoxicity-guided fractionation procedure against HL-60 leukemia cells. The chemical structure of 1 was determined by analysis of its extensive spectroscopic data. Glaziovianin A (1) displayed differential cytotoxicities in the Japanese Foundation for Cancer Research 39 cell line panel assay. The pattern of the differential cytotoxicities of 1 was found to correlate to that shown by TZT-1027, suggesting that 1 inhibited tubulin polymerization as an action mechanism. Although 1 had little influence on microtubule networks in interphase cells, 1-treated cells showed abnormal structures with unaligned chromosomes. PMID- 17428664 TI - Of Roman chariots and goats in overcoats: the syndrome of Charles Bonnet. AB - Charles Bonnet syndrome (CBS) is a widely under-recognised disorder typically characterised by complex visual hallucinations in the visually impaired. The lack of consensus over a uniform definition for CBS has much to do with the unresolved pathophysiology of the disorder. A leading hypothesis proposes that complex hallucinations arise from visual association cortical areas following their de afferentation from the central visual pathway. While treatment aimed at improving the visual deficit can limit the symptoms of CBS, at present there is no reliably effective pharmacotherapy for the disorder. Once correctly recognised, a key management principle is to reassure the patient that CBS is a well-documented clinical entity that is not a harbinger of psychiatric illness. PMID- 17428665 TI - How accurate is Whiteside's line as a reference axis in total knee arthroplasty? AB - There is an increasing focus on the precision with which prostheses in the knee are inserted with advent of computer assisted surgery. Much attention has been paid to the differences between this and conventional alignment jig techniques. Both techniques however rely on accurate identification of bony morphology and utilising this information to correctly orientate the prosthesis. Correct rotational alignment of the femoral prosthesis in total knee arthroplasty is important for correct patella tracking, patellofemoral joint contact forces, varus-valgus positioning in flexion, correct rotational alignment of the tibia in extension and the avoidance of anterior femoral notching. Whiteside's line is considered to be perpendicular to the epicondylar axis and therefore a reliable axis of reference. A cadaveric study was performed to assess the reliability and reproducibility of Whiteside's line in 50 cadaveric distal femora. Our results have shown that Whiteside's line is perpendicular to the epicondylar axis in the majority of cases (mean of 91 degrees). However the variation of values about the mean (range 80-102 degrees), and a SD of 4.7 degrees suggests that this should not be used alone as a rotational assessment guide. Rotation should ideally be checked against several axes to avoid errors in rotation positioning of the femoral prosthesis. PMID- 17428666 TI - Articular cartilage defects: study of 25,124 knee arthroscopies. AB - This retrospective study aimed to provide data on the prevalence, epidemiology and etiology of the knee articular cartilage lesions and describe and estimate, on the ground of a large database, the number of patients who might benefit from cartilage repair surgery. The analysis of 25,124 knee arthroscopies performed from 1989 to 2004 was conducted. Information concerning cartilage lesion, associated articular lesions and performed procedure were collected. Cartilage lesions were classified in accordance with the Outerbridge classification. Chondral lesions were found in 60% of the patients. Documented cartilage lesions were classified as localized focal osteochondral or chondral lesion in 67%, osteoarthritis in 29%, osteochondritis dissecans in 2% and other types in 1%. Non isolated cartilage lesions accounted for 70% and isolated lesions accounted for 30%. The patellar articular surface (36%) and the medial femoral condyle (34%) were the most frequent localization of the cartilage lesions. Grade II according to Outerbridge classification was the most frequent grade of the cartilage lesion (42%). The most common associated articular lesions were the medial meniscus tear (37%) and the injury of the anterior crucial ligament (36%). Articular cartilage lesions are a common pathology of the knee joint. The potential candidates for cartilage repair surgery, patients with one to three localized grade III and IV cartilage lesions, under the age of 40 were found in 7% and under the age of 50 years in 9% of all analysed patients. However, because these patients are a heterogeneous group and the natural history of cartilage lesions remains so far unknown, also the total number of patients in our study, who might benefit from cartilage repair, remains unknown precisely. PMID- 17428667 TI - Synthesis of andrographolide derivatives: a new family of alpha-glucosidase inhibitors. AB - Andrographolide (1), the cytotoxic agent of the plant Andrographis paniculata, was subjected to semi-synthetic studies leading to a series of new derivatives, a novel family of glucosidase inhibitors. Nicotination of 3,19-hydroxyls in 15 alkylidene andrographolide derivatives (9) was favorable to alpha-glucosidase inhibition activity. Among them, 15-p-chlorobenzylidene-14-deoxy-11,12-didehydro 3,19-dinicotinateandrographolide (11c) was a very potent inhibitor against alpha glucosidase with an IC50 value of 6 microM. However, all compounds concerned for beta-glucosidase showed no inhibition. All compounds synthesized were characterized by the analysis of NMR, IR, HRMS spectra and the stereochemistry of 2 was confirmed by X-ray analysis. PMID- 17428668 TI - The preparation and antioxidant activity of the sulfanilamide derivatives of chitosan and chitosan sulfates. AB - Chitosan (CS) and chitosan sulfates (CSS) with different molecular weight (Mw) were reacted with 4-acetamidobenzene sulfonyl chloride to obtain sulfanilamide derivatives of chitosan and chitosan sulfates (LSACS, HSACS, LSACSS, HSACSS). The preparation conditions such as different reaction time, temperature, solvent, and the molar ratio of reaction materials are discussed in this paper. Their structures were characterized by FTIR spectroscopy and elemental analyses. The antioxidant activities of the derivatives were investigated employing various established in vitro systems, such as hydroxyl-radical ((*)OH) superoxide anion (O2(*-)) scavenging and reducing power. All kinds of the compounds (HCS, LCS, HCSS, LCSS, HSACS, LSACS, HSACSS, LSACSS) showed stronger scavenging activity on hydroxyl radical than ascorbic acid (Vc). The inhibitory activities of the derivatives toward superoxide radical by the PMS-NADH system were obvious. The experiment showed that the superoxide radical scavenging effect of sulfanilamide derivatives of chitosan and chitosan sulfates was stronger than that of original CS and CSS. All of the derivatives were efficient in the reducing power. The results indicated that the sulfanilamide group were grafted on CS and CSS increased the reducing power of them obviously. PMID- 17428669 TI - Synthesis, antimicrobial, and QSAR studies of substituted benzamides. AB - A series of new substituted benzamides were synthesized and tested in vitro for their antibacterial activity against Gram-positive and Gram-negative bacteria and as well for antifungal activity. The compounds 8i and 9 showed better activity among the different benzamides synthesized. The structural characteristics governing antibacterial activities of substituted benzamides were studied using QSAR methodology. The results showed that the antimicrobial activity could be modeled using the topological descriptors, molecular connectivity indices (2chi(v) and 2chi) and Kiers shape index (kappaalpha1). The low residual activity and high cross-validated r2 values (r(cv)2) observed indicated the predictive ability of the developed QSAR models. PMID- 17428670 TI - Inhibitory effect of obovatal on the migration and invasion of HT1080 cells via the inhibition of MMP-2. AB - Because the activation of matrix metalloproteinases (MMP) is a key factor in the metastatic process, compounds with the ability to inhibit MMP activity have a potential in the treatment of tumor. From the examination of 2000 plant extracts, obovatal isolated from the extract of the leaves of Magnolia obovata THUNB was a potent inhibitor of MMP-2 enzyme in vitro. In human fibrosarcoma cells (HT1080) activated with MMP-2, obovatal inhibited MMP-2 enzyme activity and expression. In addition, the compound blocked migration and invasion of the cells. This study demonstrates that obovatal exerts its anticancer effects through blocking migration and invasion by inhibition of MMP-2 expression and activity, and also will be a good lead molecule for the development of anti-tumor drug. PMID- 17428671 TI - Quantitative diffusion tensor imaging detects dopaminergic neuronal degeneration in a murine model of Parkinson's disease. AB - Early diagnosis of Parkinson's disease (PD) is required to improve therapeutic responses. Indeed, a clinical diagnosis of resting tremor, rigidity, movement and postural deficiencies usually reflect >50% loss of the nigrostriatal system in disease. In a step to address this, quantitative diffusion tensor magnetic resonance imaging (DTI) was used to assess nigrostriatal degeneration in the 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intoxication model of dopaminergic nigral degeneration. We now demonstrate increased average diffusion (p<0.005) and decreased fractional anisotropy (p<0.03) in the substantia nigra (SN) of 5- to 7-day MPTP-treated animals when compared to saline controls. Transverse diffusivity demonstrated the most significant differences (p < or = 0.002) and correlated with the numbers of SN dopaminergic neurons (r=-0.75, p=0.012). No differences were found in the striatum, corpus callosum, cerebral cortex, or ventricles. These results demonstrate that DTI may be used as a surrogate biomarker of nigral dopaminergic neuronal degeneration. PMID- 17428672 TI - UV-Filter combinations under UV-A exposure: concomitant quantification of over all spectral stability and molecular integrity. AB - Efficient UV-absorbing molecules are designed to protect against UV-light over exposure. However, upon UV exposure they may change spectral performance or act as photooxidants via generation of free radicals and reactive oxygen species alone or in combination with others. Therefore, information about their photointegrity which comprises (i) stable absorbance and (ii) absence of UV induced molecular breakdown, is fundamental. In this study, seven commonly used UV-A, UV-B and broad spectrum UV-AB filters and their combinations, were incorporated into phosphatidylcholine (PC)-based liposomes and exposed to UV-A (275 kJ/m(2)). Spectral integrity, evaluated by recording UV-absorbance spectra of the extracted filter molecules and molecular integrity, assessed indirectly via quantification of UV-A induced PC peroxidation, revealed that spectral stability of filter molecules alone or in combination (e.g. trianilino p carboxyethylhexyl triazine, EHT plus ethylhexyl p-methoxycinnamate, OMC) does not necessarily imply absence of radical generation and that spectral lability does not necessarily have to lead to radical generation and molecular decay (e.g. OMC). This simple system capable of discriminating between essentially photostable and photounstable UV-absorbing molecules alone and in mixtures, might be useful for determining the influence of UV-protection as well as of photostability of UV-absorbers with regard to UV-induced genotoxic/phototoxic and photoageing-related, radical-based processes. PMID- 17428673 TI - Tumour necrosis factor-alpha (TNFalpha) and interleukin-10 are crucial mediators in post-operative systemic inflammatory response and determine the occurrence of complications after major abdominal surgery. AB - BACKGROUND: The course of serum cytokine levels in patients with postoperative systemic inflammatory response syndrome (SIRS) after major abdominal surgery remains currently unclear. METHODS: Blood was sampled pre- and post-operatively and on days 1 and 2 in 40 patients undergoing major abdominal surgery. Concentrations of tumour necrosis factor-alpha (TNFalpha), interleukin (IL) -6, IL-8, and IL-10 were measured by the LINCOplex assay; those of soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) by an enzyme immunoassay. RESULTS: Compared to their pre-operative values, sTREM-1 was elevated on day 2; TNFalpha on day 1; IL-6 and IL-10 post-operatively and on days 1 and 2; and IL-8 post-operatively and on day 1. The duration of operation correlated with TNFalpha and IL-10 at all sampling times, and with IL-6 post-operatively. There were no differences in cytokine concentrations between patients who exhibited post operative complications and those who did not. IL-10/TNFalpha below 30 was found in all patients with complications (100%) and in 20 patients without complications (64.5%, p: 0.043). CONCLUSIONS: SIRS following major surgery is characterised by complex alterations in cytokine concentrations. The balance between TNFalpha and IL-10 seems to determine the occurrence of post-operative complications. PMID- 17428674 TI - PPAR dual agonists: are they opening Pandora's Box? AB - Cardiovascular disorders are the major cause of mortality in patients of diabetes mellitus. Peroxisome proliferator activated receptors (PPARs) are ligand activated transcription factors of nuclear hormone receptor superfamily comprising of three subtypes such as PPARalpha, PPARgamma and PPARdelta/beta. Activation of PPARalpha reduces triglycerides and involves in regulation of energy homeostasis. Activation of PPARgamma causes insulin sensitization and enhances glucose metabolism, whereas activation of PPARdelta enhances fatty acid metabolism. Current therapeutic strategies available for the treatment of diabetes do not inhibit the associated secondary cardiovascular complications. Hence, the development of multimodal drugs which can reduce hyperglycemia and concomitantly inhibit the progression of secondary cardiovascular complications may offer valuable therapeutic option. Several basic and clinical studies have exemplified the beneficial effects of PPARalpha and PPARgamma ligands in preventing the cardiovascular risks. The PPARalpha/gamma dual agonists are developed to increase insulin sensitivity and simultaneously prevent diabetic cardiovascular complications. Such compounds are under clinical trials and proposed for treatment of Type II diabetes with secondary cardiovascular complications. However, PPARalpha/gamma dual agonists such as muraglitazar, tesaglitazar and ragaglitazar have been noted to produce several cardiovascular risks and carcinogenicity, which raised number of questions about the clinical applications of dual agonists in diabetes and its associated complications. The ongoing basic studies have elucidated the cardio protective role of PPARdelta. Therefore, further studies are on the track to develop PPARalpha/delta and PPAR gamma/delta dual agonists and PPARalpha/gamma/delta pan agonists for the treatment of diabetic cardiovascular complications. The present review critically analyzes the protective and detrimental effect of PPAR agonists in diabetic cardiovascular complications. Moreover, the newly developed PPARalpha/delta and PPAR gamma/delta dual agonists and PPARalpha/gamma/delta pan agonists have also been discussed which may open a new vista in the management of diabetic cardiovascular complications in near future. PMID- 17428676 TI - BDNF regulates the expression and traffic of NMDA receptors in cultured hippocampal neurons. AB - The neurotrophin BDNF regulates the activity-dependent modifications of synaptic strength in the CNS. Physiological and biochemical evidences implicate the NMDA glutamate receptor as one of the targets for BDNF modulation. In the present study, we investigated the effect of BDNF on the expression and plasma membrane abundance of NMDA receptor subunits in cultured hippocampal neurons. Acute stimulation of hippocampal neurons with BDNF differentially upregulated the protein levels of the NR1, NR2A and NR2B NMDA receptor subunits, by a mechanism sensitive to transcription and translation inhibitors. Accordingly, BDNF also increased the mRNA levels for NR1, NR2A and NR2B subunits. The neurotrophin NT3 also upregulated the protein levels of NR2A and NR2B subunits, but was without effect on the NR1 subunit. The amount of NR1, NR2A and NR2B proteins associated with the plasma membrane of hippocampal neurons was differentially increased by BDNF stimulation for 30 min or 24 h. The rapid upregulation of plasma membrane associated NMDA receptor subunits was correlated with an increase in NMDA receptor activity. The results indicate that BDNF increases the abundance of NMDA receptors and their delivery to the plasma membrane, thereby upregulating receptor activity in cultured hippocampal neurons. PMID- 17428677 TI - Overexpression in Escherichia coli and functional reconstitution of the liposome binding ferriheme protein nitrophorin 7 from the bloodsucking bug Rhodnius prolixus. AB - A number of ferriheme proteins, termed nitrophorins (NPs), occur in the saliva of the bloodsucking insect Rhodnius prolixus ('kissing bug'), which is a vector for Chagas' disease. Nitrophorins bind the heme b cofactor in the beta-barrel of their lipocalin fold, which is further anchored through a proximal histidine Fe(III) bond. The distal Fe(III) coordination site then binds nitric oxide (NO) for delivery into a host's tissues during blood feeding, where, upon NO release, the distal Fe(III) site acts as a histamine trap to delay the victim's immune response. Previously, four nitrophorins from R. prolixus, NP1 to NP4, have been extensively characterized. Recently, another nitrophorin, NP7, was discovered in a cDNA library derived from the same insect. Among the R. prolixus nitrophorins, NP7 was found to be unique in its ability to bind to negatively charged cell surfaces. However, the yield of functional recombinant NP7 was rather low when the established protocol for NP1-4 was followed. Here, we report on a novel expression and reconstitution method for NP7 that yields sufficient amounts of pure protein for extensive characterization (28-fold increase). This method may prove useful for the reconstitution of other proteins with a lipocalin fold. PMID- 17428678 TI - Expression and purification of the minor histocompatibility antigen, HA-1H generated in Escherichia coli. AB - The minor histocompatibility antigen HA-1H is a potential immunotherapeutic molecule. It can be used as a target for graft versus leukaemia reactions to eliminate residual HA-1H expressing leukaemic cells in patients following haemopoietic stem cell transplantation, whereby HA-1H primed donor cells can be transferred into a patient via adoptive immunotherapy. However, thus far only synthetic peptides corresponding to a HLA-A *0201 restricted HA-1H epitope have been used to generate HA-1H specific T cells. We are the first laboratory to clone, express and purify a region of HA-1H using an Escherichia coli expression system. The recombinant HA-1H protein was purified under denaturing conditions and the affinity purification tag removed using thrombin to remove non-specific amino acids. The 92 amino acid recombinant protein was characterised by mass spectrometry. Our rationale is that by using a recombinant HA-1H protein rather than peptide, HA-1H specific T cells may be generated from presentation of multiple HA-1H epitopes complexed in different HLA molecules. A multi-epitope approach may have wider applicability and maybe more effective at leukaemia control. The recombinant HA-1H protein may also be used as a research tool to identify novel CD4(+) helper T cell and CD8(+) cytotoxic T cell epitopes. PMID- 17428679 TI - PML protein association with specific nucleolar structures differs in normal, tumor and senescent human cells. AB - Promyelocytic leukemia protein (PML), a tumor suppressor, forms in most human cell types discrete multiprotein complexes termed PML nuclear bodies. Here, we have used indirect immunofluorescence and confocal microscopy to describe various forms of a novel nuclear PML compartment associated with nucleoli that is found under growth-permitting conditions in human mesenchymal stem cells (hMSC) and skin fibroblasts but not in several immortal cell lines with defects in the p53 and pRb pathways. In addition, we found that shut-off of rRNA synthesis induced by actinomycin D causes PML translocation to the surface of segregated nucleoli. This translocation is dynamic and reversible, following changes in nucleolar activity. Intriguingly, treatment causing premature senescence restores PML binding to nucleoli-derived structures and to the surface of segregated nucleoli in HeLa cells. These findings indicate that PML may be involved in nucleolar functions of normal non-transformed or senescent cells. The absence of nucleolar PML compartment in rapidly growing tumor-derived cells suggests that PML association with the nucleolus might be important for cell-cycle regulation. PMID- 17428680 TI - Detecting molecular interactions that stabilize, activate and guide ligand binding of the sodium/proton antiporter MjNhaP1 from Methanococcus jannaschii. AB - Integral membrane proteins are involved in virtually every cellular process. Precisely regulating these machineries would allow controlling many human and vertebrate diseases. Embedded into cellular membranes, membrane proteins establish molecular interactions that sensitively react to environmental changes and to molecular compounds, such as ligands or inhibitors. We applied atomic force microscopy (AFM) to image the Na(+)/H(+) antiporter MjNhaP1 from Methanococcus jannaschii, and single-molecule force spectroscopy (SMFS) to probe molecular interactions that drive the protein structure-function relationship. High-resolution AFM topographs showed the dimeric assembly of MjNhaP1 being reconstituted into a lipid bilayer. SMFS of MjNhaP1 unraveled molecular interactions stabilizing individual structural domains. Transmembrane domains exhibited certain probabilities to unfold individually or cooperatively with other domains resulting in different unfolding pathways. Helices VIII and X established pH sensitive interactions altering significantly upon MjNhaP1 activation, while removal of the ligand (Na(+)) destabilized the entire antiporter except helix VIII. It is assumed that Asp234/235 of helix VIII are involved in the ligand-binding site and that helix X plays a functional role in the activation of the transporter. PMID- 17428681 TI - Dissociation and unfolding of bovine odorant binding protein at acidic pH. AB - Dissociation of bovine odorant binding protein (bOBP) dimers to monomers at pH 2.5 has been confirmed through size exclusion chromatography experiments. Moreover, structural and binding properties of the acidic monomer and neutral dimer have been compared using a combination of experimental (circular dichroism and fluorescence) and computational (molecular dynamics) techniques. The secondary and tertiary structures of bOBP are largely maintained at acidic pH, but molecular dynamics simulations suggest the loop regions (N-terminal residues, Omega-loop and C-terminal segments) are more relaxed and Phe36 and Tyr83 residues are involved in the regulation of the binding cavity entrance. The formation of a molten globule state at acidic pH, suggested by the strong enhancement of fluorescence of 8-anilino-1-naphtalenesulphonic acid (ANS), is not confirmed by any significant change in the near UV circular dichroism spectrum. Functionality measurements, deduced from the interaction of bOBP with 1-amino-anthracene (AMA), show that the binding capacity of the protein at acidic pH is preserved, though slightly looser than at neutral pH. Unfolding of acidic bOBP, induced by guanidinium chloride (GdnHCl), was investigated by means of CD spectroscopy, steady state fluorescence, fluorescence anisotropy and light scattering. The stability of the acidic monomer is lower than that of the neutral dimer, owing to the loss of the swapping interactions, but renaturation is completely reversible. Finally, in contrast with the neutral dimer, at low denaturant concentration some aggregation of the acidic monomer, which vanishes before the unfolding transition, has been observed. PMID- 17428682 TI - Molecular cloning and characterization of a short type peptidoglycan recognition protein (CfPGRP-S1) cDNA from Zhikong scallop Chlamys farreri. AB - Peptidoglycan recognition protein (PGRP) specifically binds to peptidoglycan and plays a crucial role in the innate immune responses as a pattern recognition receptor (PRR). The cDNA of a short type PGRP was cloned from scallop Chlamys farreri (named CfPGRP-S1) by homology cloning with degenerate primers, and confirmed by virtual Northern blots. The full length of CfPGRP-S1 cDNA was 1073 bp in length, including a 5' untranslated region (UTR) of 59 bp, a 3' UTR of 255 bp, and an open reading frame (ORF) of 759 bp encoding a polypeptide of 252 amino acids with an estimated molecular mass of 27.88 kDa and a predicted isoelectric point of 8.69. BLAST analysis revealed that CfPGRP-S1 shared high identities with other known PGRPs. A conserved PGRP domain and three zinc-binding sites were present at its C-terminus. The temporal expression of CfPGRP-S1 gene in healthy, Vibrio anguillarum-challenged and Micrococcus lysodeikticus-challenged scallops was measured by RT-PCR analysis. The expression of CfPGRP-S1 was upregulated initially in the first 12 h or 24 h either by M. lysodeikticus or V. anguillarum challenge and reached the maximum level at 24 h or 36 h, then dropped progressively, and recovered to the original level as the stimulation decreased at 72 h. There was no significant difference between V. anguillarum and M. lysodeikticus challenge. The results indicated that the CfPGRP-S1 was a constitutive and inducible acute-phase protein which was involved in the immune response against bacterial infection. PMID- 17428683 TI - Human brain activation during phonation and exhalation: common volitional control for two upper airway functions. AB - Phonation is defined as a laryngeal motor behavior used for speech production, which involves a highly specialized coordination of laryngeal and respiratory neuromuscular control. During speech, brief periods of vocal fold vibration for vowels are interspersed by voiced and unvoiced consonants, glottal stops and glottal fricatives (/h/). It remains unknown whether laryngeal/respiratory coordination of phonation for speech relies on separate neural systems from respiratory control or whether a common system controls both behaviors. To identify the central control system for human phonation, we used event-related fMRI to contrast brain activity during phonation with activity during prolonged exhalation in healthy adults. Both whole-brain analyses and region of interest comparisons were conducted. Production of syllables containing glottal stops and vowels was accompanied by activity in left sensorimotor, bilateral temporoparietal and medial motor areas. Prolonged exhalation similarly involved activity in left sensorimotor and temporoparietal areas but not medial motor areas. Significant differences between phonation and exhalation were found primarily in the bilateral auditory cortices with whole-brain analysis. The ROI analysis similarly indicated task differences in the auditory cortex with differences also detected in the inferolateral motor cortex and dentate nucleus of the cerebellum. A second experiment confirmed that activity in the auditory cortex only occurred during phonation for speech and did not depend upon sound production. Overall, a similar central neural system was identified for both speech phonation and voluntary exhalation that primarily differed in auditory monitoring. PMID- 17428684 TI - Cerebral activation in patients with somatoform pain disorder exposed to pain and stress: an fMRI study. AB - Patients with somatoform pain disorders are supposed to suffer from an early acquired defect in stress regulation. In order to look for common alterations of the pain- and stress-responsive cortical areas, we prospectively recorded cerebral activations induced by pin-prick pain, by cognitive stress and emotional stress using functional magnetic resonance imaging (fMRI) in a group of 17 patients and an age-matched control group. In addition, the hippocampal volumes of both groups were measured. Patients showed increased activations of the known pain-processing areas (thalamus, basal ganglia, operculo-insular cortex), but also of some prefrontal, temporal and parietal regions during first pain exposure and of temporal and parietal areas during cognitive stress, but reduced activations during emotional stress. In contrast to these functional differences, hippocampal volume was not significantly reduced in patients. Although the superior temporal gyrus was the only common area of an "overactivation" in patients in the pain and stress condition, findings of our study support the current concept of mechanisms involved in somatoform pain disorders: central processing of pain and of cognitive stress is increased in patients possibly due to exaggerated memory and/or anticipation of pain exposure and to a disturbance of stress-regulating systems which has to be worked out on a cortical level in more detail. Our finding of a reduced responsiveness to emotional stress is surprising, but not contradictive to these results because some sort of neglect or coping mechanisms may have developed over time as a response to early adversities. PMID- 17428685 TI - Dynamics of hemispheric activity during metaphor comprehension: electrophysiological measures. AB - Brain imaging studies have lead to conflicting findings regarding the involvement of the right hemisphere (RH) in metaphor comprehension. Some report more relative RH activation when processing figurative expressions but others have shown just the opposite. The inconsistencies might be a result of the low temporal resolution related to current brain imaging techniques which is insufficient to uncover patterns of hemispheric interaction that change over time. Event-related potentials and a source estimation technique (LORETA) were used to investigate such temporal interactions when processing two-word expressions denoting literal, conventional metaphoric, and novel metaphoric meaning, as well as unrelated word pairs. Participants performed a semantic judgment task in which they decided whether each word pair conveyed a meaningful expression. Our findings indicate that during comprehension of novel metaphors there are some stages of considerable RH involvement, mainly of the temporal and superior frontal areas. Although the processing mechanisms used for all types of expressions were similar and require both hemispheres, the relative contribution of each hemisphere at specific processing stages depended on stimulus type. Those stages correspond roughly to the N400 and LPC components which reflect semantic and contextual integration, respectively. The present study demonstrates that RH mechanisms are necessary, but not sufficient, for understanding metaphoric expressions. Both hemispheres work in concert in a complex dynamical pattern during literal and figurative language comprehension. Electrophysiological recordings together with source localization algorithms such as LORETA are a viable tool for measuring this type of activity patterns. PMID- 17428686 TI - Functional magnetic resonance imaging: measuring versus estimating. AB - Brain imaging techniques largely spread in neuroscience literature. Due to initial technical limitations such as the very low signal-to-noise ratio, group experiments became the rule. This fact, together with the wide use of standard brains to localize the activations, lead several experimenters to the wrong idea that the brain can be described by a Cartesian coordinate system, neglecting at the same time the importance of individual neuroanatomy. My commentary on the paper by Devlin and Poldrack reinforces their reminder that it is necessary to deal with anatomy. Moreover, it adds some considerations on the relevance of single subjects studies and on the importance of the BOLD intensity signal, which should be used to describe brain activity together with the most used statistical tools. PMID- 17428687 TI - Automatic detection and quantification of hippocampal atrophy on MRI in temporal lobe epilepsy: a proof-of-principle study. AB - In temporal lobe epilepsy (TLE), hippocampal atrophy (HA) is a marker of poor prognosis regarding seizure remission, but predicts success of anterior temporal lobe resection. Manual quantification of HA on MRI is time-consuming and limited by investigator availability. Normal ranges of hippocampal volumes, both in absolute terms and relative to intracranial volume, and of hippocampal asymmetry were defined using an automatic label propagation and decision fusion technique based on thirty manually derived atlases of healthy controls. Manual test-retest reliability and overlaps of automatically and manually determined hippocampal volumes were quantified with similarity indices (SIs). Correct clinical identification of ipsilateral HA, and contralaterally normal hippocampal volumes, was determined in nine patients with histologically confirmed hippocampal sclerosis in terms of volumes and asymmetry indices (AIs) for standard statistical thresholds and with receiver operating characteristic (ROC) analysis. Manual test-retest reliability was very high, with SIs between 0.87 and 0.90. Manual and automatic hippocampus labels overlapped with a SI of 0.83 on the unaffected but with 0.76 on the atrophic side. Accuracy was higher for less atrophic hippocampi. The automatic method correctly identified 6/9 HAs in terms of absolute volume, 7/9 in terms of relative volume at a standard 2 SD threshold, and 9/9 for AIs. ROC-determined thresholds allowed clinically desirable correct identification of all HAs (100% sensitivity) with 85-100% specificity for volumes, and 100% specificity for AIs. The method has the potential to automatically detect unilateral HA, but further work is needed to determine its performance in detecting clinically important bilateral disease. PMID- 17428688 TI - Phylogeography of rusty-necklaced partridge (Alectoris magna) in northwestern China. AB - Rusty-necklaced partridges (Alectoris magna) are found in Qinghai, Ningxia and Gansu provinces of northwestern China, distributed in mountainous areas that were heavily affected by cyclical climate changes throughout the Pleistocene. We investigated the potential impact of Pleistocene range shifts on phylogeographic patterns using 458 nucleotides of mitochondrial control-region from 102 individuals sampled from 10 populations across the current range. We found 31 haplotypes defined by 33 polymorphic positions. Phylogenetic analyses revealed two robustly supported clades, one corresponding to the Chaidamu Basin and the other including all individuals from all other populations. At the population level we also found significant evidence of differentiation. The phylogeographic structure of the rusty-necklaced partridge is consistent with the "phylogenetic discontinuity, spatial separation" pattern outlined by Avise et al. [Avise, J.C., Arnold, J., Ball, R.M., Bermingham, E., Lamb, T., Niegel, J.E., Reeb, C.A., Saunders, N.C., 1987. Intraspecific phylogeography: the mitochondrial DNA bridge between population genetics and systematics. Ann. Rev. Ecol. Syst. 18, 489-522]. PMID- 17428689 TI - Health-related quality of life in patients with osteoarthritis after total knee replacement: factors influencing outcomes at 36 months of follow-up. AB - OBJECTIVES: (1) To evaluate health-related quality of life (HRQL) in patients with severe osteoarthritis (OA) undergoing total knee replacement (TKR) and (2) to identify the influence of sociodemographic, clinical, intra-operative and postoperative variables on HRQL at 36 months after TKR. DESIGN: Prospective study with a 36-month follow-up. Preoperative interviews were carried out with 90 in patients. The disease-specific Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaire was used to measure the health status. Sociodemographic, clinical, intra-operative degree of difficulty, in-patient and postoperative data were collected. Associations were analyzed using linear regression models. RESULTS: Of the 90 potentially eligible patients, 67 (54 females, mean age 74.83, standard deviation [SD] 5.57) completed follow-up assessment. There were significant differences between preoperative and postoperative WOMAC pain, stiffness and function scores (P<0.001, P=0.005 and P<0.001, respectively). Variables retained in each of the models explained between 15% and 23% (R(2) adjusted) of the variability of each WOMAC dimension. Higher preoperative WOMAC scores were associated with greater postoperative improvement (P<0.001). Chronic musculoskeletal pain unrelated to knee OA was associated with higher WOMAC pain, stiffness and function dimension scores (P=0.004, P=0.029 and P=0.005, respectively). Severe (Class III) obesity (body mass index [BMI] 35-39.9) was associated with more pain (P=0.049). CONCLUSIONS: In patients with severe OA, HRQL significantly improved at 36 months after TKR, especially in the pain dimension. Lower preoperative WOMAC scores, chronic pain unrelated to knee OA, and severe obesity negatively influenced postoperative WOMAC scores. This disease-specific questionnaire may help to identify patients at increased risk of negative outcomes after surgery. PMID- 17428690 TI - Detection of potassium currents and regulation of multidrug resistance by potassium channels in human gastric cancer cells. AB - The present study aimed to investigate the potassium currents and further explore the role of potassium channels in drug response of gastric cancer cells. By patch clamp technique, potassium currents of human gastric cancer cell SGC7901 were recorded in the mode of voltage clamp. Both 4-aminopyridine (4-AP) and tetraethylammonium (TEA) could almost completely block this current. The chemotherapeutic drugs, adriamycin or 5-fluorouracil could significantly increase the K(+) current density on SGC7901 cells in a dose-dependent manner. 4-AP or TEA was found to restrain adriamycin-induced apoptosis and enhance multidrug resistant phenotype of SGC7901 cells. Up-regulation of Kv1.5, which has been found widely expressed in gastric cancer cells including SGC7901, increased the K(+) current density and sensitivity of SGC7901 cells to multiple chemotherapeutic drugs, whereas down-regulation of Kv1.5 enhanced the drug resistant phenotype of SGC7901 cells. In conclusion, potassium channels may exert regulatory effects on multidrug resistance by regulating drug-induced apoptosis in gastric cancer cells. PMID- 17428691 TI - Involvement of the cytoskeleton in the secretory pathway and plasma membrane organisation of higher plant cells. PMID- 17428692 TI - Strong association of interleukin-4 (-590 C/T) polymorphism with increased risk for oral squamous cell carcinoma in Europeans. AB - OBJECTIVE: The present study was performed to investigate the possible association of -590 C/T polymorphism in the interleukin-4 (IL-4) gene which affects its expression with the risk for development of oral cancer. STUDY DESIGN: Polymerase chain reaction-based restriction analysis was performed in DNA samples of 156 patients with oral squamous cell carcinoma (OSCC) and 162 healthy control subjects of equivalent gender, age, and ethnicity (Greek and German). Statistical analyses were performed conducting Fisher exact test. RESULTS: The T/T genotype was associated with an increased risk for the development of OSCC (P = .018; OR 1.47, 95% CI 0.66-3.28), especially for early stages of this malignancy (P < .0001; OR 3.17, 95% CI 1.31-7.65). CONCLUSIONS: The above findings are consistent with the growth-promoting role of IL-4 in head and neck cancer and its inhibitory effect on neoangiogenesis and metastasis. The present study in Europeans is not in accordance with a previous report of unclear association of this polymorphism in a Chinese population. PMID- 17428693 TI - Intraoral granulocytic sarcoma presenting as multiple maxillary and mandibular masses: a case report and literature review. AB - Granulocytic sarcoma (GS) is an unusual localized tumor composed of immature granulocytic precursor cells that occurs in extramedullary sites. However, GS involving the oral cavity is rare. We report a case of intraoral GS with an unusual clinical presentation, including a history of chronic myelogenous leukemia in remission, multiple maxillary and mandibular gingival masses mimicking acute inflammation that developed over a short period, complete remission after 1 week of treatment with imatinib mesylate (Gleevec), and no bone marrow or peripheral blood involvement over a 6-month follow-up period. To our knowledge, this is the first report of treatment of intraoral GS with Gleevec resulting in a complete remission. PMID- 17428694 TI - Diagnosing osteoporosis by using dental panoramic radiographs: the OSTEODENT project. AB - OBJECTIVES: Measurement of cortical thickness and subjective assessment of cortical porosity on panoramic radiographs are methods previously reported for diagnosing osteoporosis. The aims of this study were to determine the relative efficacy of the mandibular cortical index and cortical width in detecting osteoporosis, both alone and in combination, and to determine the optimal cortical width threshold for referral for additional osteoporosis investigation. STUDY DESIGN: Six hundred seventy-one postmenopausal women 45 to 70 years of age were recruited for this study. They received dual energy x-ray absorptiometry (DXA) scans of the left hip and lumbar spine (L1 to L4), and dental panoramic radiographic examinations of the teeth and jaws. Three observers separately assessed the mandibular cortical width and porosity in the mental foramen region of the mandible. Cortical width was corrected for magnification errors. Chi squared automatic interaction detection analysis (CHAID) software was used (SPSS AnswerTree, version 3.1, SPSS Inc., Chicago, IL). RESULTS: Chi-squared automatic interaction detection analysis showed that the cortical porosity was a poorer predictor of osteoporosis than mandibular cortical width. For the 3 observers, a mandibular cortical width of <3 mm provided diagnostic odds ratios of 6.51, 6.09, and 8.04. The test is therefore only recommended in triage screening of individuals by using radiographs made for purposes other than osteoporosis. CONCLUSION: When evaluating panoramic radiographs, only those patients with the thinnest mandibular cortices (i.e., <3 mm) should be referred for further osteoporosis investigation. PMID- 17428695 TI - The effect of thermal treatment on the resistance of nickel-titanium rotary files in cyclic fatigue. AB - OBJECTIVE: The purpose of this study was to determine the effect of various thermal treatments on the fatigue resistance of a nickel-titanium (NiTi) engine driven endodontic file. STUDY DESIGN: Fifteen groups of 5 files each of ISO 30 and taper .04 were tested in this study. The cutting tip (5 mm from the end) of files from 14 groups were heat treated for 30 minutes in temperatures 250 degrees C, 300 degrees C, 350 degrees C, 375 degrees C, 400 degrees C, 410 degrees C, 420 degrees C, 425 degrees C, 430 degrees C, 440 degrees C, 450 degrees C, 475 degrees C, 500 degrees C, and 550 degrees C, respectively, while 1 group was used as reference. The files were placed in a device that allowed the instruments to be tested for rotating bending fatigue inside an artificial root canal. The number of rotations to breakage was recorded for each file. The mean values of all groups were statistically analyzed using 1-way analysis of variance and Student Newman Keuls multiple comparison test at alpha = .05. RESULTS: The 430 degrees C and 440 degrees C groups showed the highest values, with fatigue resistance decreasing for thermal treatment at lower and higher temperatures. This may be the result of metallurgical changes during annealing. CONCLUSION: Within the limitations of the low sample size and the specific instrument size tested, it appears that the appropriate thermal treatment may significantly increase the fatigue resistance of the NiTi file tested. PMID- 17428696 TI - Alteration of the mandibular plane during sagittal split advancement: short- and long-term stability. AB - OBJECTIVES: We investigated predictors of long- and short-term stability of surgical mandibular advancements with bilateral sagittal split osteotomy (BSSO). STUDY DESIGN: Class II patients (n = 127) received mandibular advancement through BSSO with either rigid internal fixation or wire osteosynthesis. We used multiple linear regression analysis to assess the association of predictor variables with post-treatment horizontal and vertical B-point movement through 2 years. RESULTS: Counterclockwise rotation of the mandibular plane angulation (MPA) was associated with greater horizontal and vertical relapse at all time periods except 8 weeks. Wire osteosynthesis, larger advancements, younger age, and genioplasty were significantly associated with relapse. CONCLUSIONS: Surgically closing the MPA is associated with late horizontal and vertical relapse, whereas fixation type is related to early B-point movement. Large advancements with forward and upward repositioning of the mandible, genioplasty, and young age also play a role in relapse after BSSO. PMID- 17428697 TI - Peripheral and central giant cell granulomas of the jaws: a demographic study. AB - OBJECTIVE: This study aimed to assess the demographic characteristics of peripheral giant cell granulomas (PGCGs) and central giant cell granulomas (CGCGs) in patients treated at our centers. STUDY DESIGN: This 12-year retrospective study was based on existing data. Files of patients from 1993-2004 with a definite diagnosis of PGCGs and CGCGs from the oral pathology departments of our universities were assessed. Information regarding age distribution, gender, the jaw involved, the presenting area of the lesion, surgical treatment, and recurrence was documented. RESULTS: During the study period, 204 patients with CGCGs were treated. The patients with CGCGs varied in age from 5 to 72 years, and the mean age patients was 23.72 years. Among these, 127 cases (62.87%) occurred in the second and third decades of life. One hundred thirty cases (63.75%) occurred in females and 74 (36.25%) in males. Ninety cases (44.1%) presented in posterior parts of the jaws. One hundred forty-four cases (70.58%) appeared in the mandible. Peripheral GCGs presented in 575 patients, who varied in age from 2 to 85 years with a mean age of 31.02 years. Among these, 297 cases (51.65%) occurred in females and 278 (48.34%) in males. Four hundred sixty-seven cases (81.2%) occurred in the first five decades of life, and 352 cases (61.21%) appeared in the mandible. CONCLUSIONS: Giant cell granulomas comprised 9.29% of oral lesions. Peripheral GCG lesions occurred more than 2 times more frequently than CGCGs. Central GCGs occurred about 2 times more frequently in females, whereas PGCGs had an equal prevalence in both genders (P < .05). The mean age for patients with CGCGs was less than patients with PGCGs (P < .05). Central GCGs involved the mandible approximately 2 times more frequently than the maxilla (P < .05). However, when presenting in the maxilla, CGCGs most frequently presented in the area anterior to the canines (P < .05). Peripheral GCGs involved the mandible approximately 1.5 times more frequently than the maxilla (P < .05). Thorough curettage was the main treatment modality used. There were 9 cases (4.41%) of recurrence of CGCGs and 8 cases (1.39%) of recurrence of PGCGs documented during the follow-up period (ranging from 1-12 years). PMID- 17428698 TI - Comparison of corticocancellous block and particulate bone grafts in maxillary sinus floor augmentation for bone healing around dental implants. AB - OBJECTIVE: The aim of this study was to compare 2 types of bone used for maxillary sinus floor augmentation, corticocancellous block or particulate bone grafts, on bone healing around dental implants when installed simultaneously with the implant. STUDY DESIGN: The mucous membranes of 12 sinuses in 6 dogs were elevated bilaterally. On one side of the maxillary sinus, autogenous corticocancellous block bone was grafted into the space between the membrane and sinus wall. On the opposite side, autogenous corticocancellous particulate bone was grafted. Simultaneously, 2 dental implants were inserted into the grafting material through the maxillary sinus floor. The animals were killed 6 months after surgical procedure. RESULTS: The mean bone-implant contact was 56.7% on the block side and 32.1% on the particulate side (P < .05). The mean height of newly formed bone in the augmented area was 12.3 mm on the block side and 9.7 mm on the particulate side (P < .05). CONCLUSION: Our results show that maxillary sinus floor augmentation using corticocancellous block bone grafts, when installed simultaneously with the implant, is superior to corticocancellous particulate bone grafts for bone healing around dental implants. PMID- 17428699 TI - Surgery of lymphatic malformations in oral and cervicofacial regions in children. AB - OBJECTIVE: The objective of this study was to determine prognostic factors and complication rates related to the surgical management of lymphatic malformations of oral and cervicofacial regions in children. STUDY DESIGN: The charts of 117 children operated on for oral and cervicofacial lymphatic malformations were retrospectively reviewed. Treatment outcomes were analyzed for correlation with several factors, including age at presentation, sex, associated symptoms, anatomical site of involvement, extent of disease, operative complications, histological pattern, and recurrence. The chi(2) test was used to compare treatment failure rates and complication rates between patients with and without these factors. RESULTS: The tongue was the most frequent site of involvement (40.17%). The lesions with oral and facial involvement had a higher failure or recurrence rate (29.23%) than those with cervical involvement (8.33%; P < .05), and lesions with involvement of multiple sites had a higher recurrence rate (48.28%) than those with involvement of a single site or 2 sites (11.67%; P < .01). Although patients who underwent surgical procedure at less than 1 year of age had a higher recurrence rate (28.95%) than those more than 1 year (19.61%) of age, and microcystic lesions had a higher recurrence rate (28.33%) than macrocystic lesions (13.79%), no significant difference was found between them (P > .05). The lesions with involvement of 3 or more sites had significantly higher operative complication rates (37.8%) than those with involvement of 1 or 2 sites (15.58%; P < .01). CONCLUSION: Factors correlated with a worse prognosis in lymphatic malformations of oral and cervicofacial regions include the involvement of the oral cavity and/or face and involvement of multiple anatomical sites, which also may be associated with higher operative complications. PMID- 17428700 TI - Changes in the temporomandibular joint after mandibular setback surgery in monkeys: intraoral vertical versus sagittal split ramus osteotomy. AB - OBJECTIVE: This study was to investigate the changes in condylar position and structure of the temporomandibular joint (TMJ) after mandibular setback using 2 forms of the ramus osteotomy. STUDY DESIGN: Twelve adult male rhesus monkeys were randomly divided into groups A (n = 6) and B (n = 6) for mandibular setback surgical procedure. An intraoral vertical ramus osteotomy (IVRO) was performed in group A, whereas a sagittal split ramus osteotomy (SSRO) was performed in group B. Changes in condylar position were quantified by computed tomography (CT) preoperatively and postoperatively. All animals in groups A and B were killed at 12 weeks after surgical procedure. The TMJ specimens were harvested and processed for histological examination. RESULTS: In group A, the CT examinations showed a significant anteroinferior displacement of the condyle after surgical procedure. In group B, slight posterior displacement and lateral tilting of the condyle were noted after surgical procedure. Thickened cartilage layer and endochondral ossification were seen in the condyles of group A, but only minimal alteration in articular cartilage was found in group B. CONCLUSIONS: Both SSRO and IVRO can be biologically sound procedures for correction of mandibular prognathism. Intraoral vertical ramus osteotomy procedure leads to condylar adaptive remodeling, which may have favorable effects on TMJ, and it could be considered as a preferred surgical treatment for those patients with preoperative TMJ disorders. PMID- 17428701 TI - Peripheral alcohol injections in the management of trigeminal neuralgia. AB - OBJECTIVE: To assess the effectiveness of peripheral alcohol injections in the management of trigeminal neuralgia. STUDY DESIGN: Retrospective case audit of patients who received peripheral alcohol injections in 1994-1999 was conducted. We examined for occurrence and duration of effect of alcohol blocks, the effect of age and repeated administration, and the effect of their use on other surgical treatments. RESULTS: Peripheral alcohol injections lasted for a mean of 11 months. Their effectiveness and complication rates were not affected by age or repeated administration. Their use did not affect, nor was their effectiveness affected by, the use of other surgical treatments. CONCLUSION: Peripheral alcohol injections continue to have a role in the management of trigeminal neuralgia. They are particularly useful in those refractory to medical management and in those who are unable or unwilling to undergo neurosurgical treatment. Peripheral alcohol injections also have a role in those awaiting neurosurgical opinions or treatment. PMID- 17428702 TI - Overexpression of HFE in HepG2 cells reveals differences in intracellular distribution and co-localization of wt- and mutated forms. AB - Liver is the primary target organ of Hereditary Hemochromatosis Type I, with the HFE mutations C282Y and H63D recognized as markers of this iron-overload disease. Hepatocytes are also the main site of synthesis of HFE. However, most early studies of overexpression of HFE were done in non-hepatic, non-HFE-expressing, cell lines. Here we report the setting up of a stable transfection model of wt- and mutant-HFE (H63D and C282Y) proteins in a hepatic cell line (HepG2), the analysis of its intracellular distribution and the effect of diferric transferrin on HFE localization. The C282Y mutant is retained in the ER, whereas HFE-wt and H63D co-localize with TfR1 exclusively in early recycling endosomes. Holotransferrin induces a re-localization of wt- and H63D-HFE, from early recycling endosomes to the cytoplasmic membrane. In conclusion our results establish the HepG2 cell line as a valuable model for the study of HFE. PMID- 17428703 TI - Transferrin receptor 2 is frequently expressed in human cancer cell lines. AB - Different proteins ensure the fine control of iron metabolism at the level of various tissues. Among these proteins, it was discovered a second transferrin receptor (TfR2), that seems to play a key role in the regulation of iron homeostasis. Its mutations are responsible for type 3 hemochromatosis (Type 3 HH). Although TfR2 expression in normal tissues was restricted at the level of liver and intestine, we observed that TfR2 was frequently expressed in tumor cell lines. Particularly frequent was its expression in ovarian cancer, colon cancer and glioblastoma cell lines; less frequent was its expression in leukemic and melanoma cell lines. Interestingly, in these tumor cell lines, TfR2 expression was inversely related to that of receptor 1 for transferrin (TfR1). Experiments of in vitro iron loading or iron deprivation provided evidence that TfR2 is modulated in cancer cell lines according to cellular iron levels following two different mechanisms: (i) in some cells, iron loading caused a downmodulation of total TfR2 levels; (ii) in other cell types, iron loading caused a downmodulation of membrane-bound TfR2, without affecting the levels of total cellular TfR2 content. Iron deprivation caused in both conditions an opposite effect compared to iron loading. These observations suggest that TfR2 expression may be altered in human cancers and warrant further studies in primary tumors. Furthermore, our studies indicate that, at least in tumor cells, TfR2 expression is modulated by iron through different biochemical mechanisms, whose molecular basis remains to be determined. PMID- 17428705 TI - Placental transfer of etomidate in pregnant ewes after an intravenous bolus dose and continuous infusion. AB - Etomidate (ETO) is a short-acting intravenous (IV) anaesthetic characterised by cardiopulmonary stability and favourable pharmacokinetics. Although ETO has been used satisfactorily in obstetrical anaesthesia, little is known about placental transfer and the drug's pharmacokinetics in the fetus. Placental transfer in pregnant ewes has been evaluated following the administration of an IV bolus of 1mg/kg ETO; and after a 1-h infusion of 100 microg/kg min(-1) ETO preceded by an IV bolus of 1mg/kg. In ewes, ETO concentration and AUC were higher than those found in fetuses. After the ETO bolus dose, the fetus:ewe AUC ratio was 0.45+/ 0.32, and the mean residence time (MRT) was 20+/-7 min for dams and 22+/-3 min for the fetuses. After ETO infusion, the AUC ratio was 0.37+/-0.08, and MRT was 46+/-12 min for ewes and 46+/-22 min for fetuses. Although ETO crosses the placenta very rapidly and reaches the fetus in high amounts, a certain placental barrier effect limits its transfer. There is no evidence of cumulative effects of the drug in the fetus as fetal ETO elimination was as rapid as in the dam. PMID- 17428706 TI - Does the assessment of general movements without video observation reliably predict neurological outcome? AB - OBJECTIVE: To assess the clinical value of a modified version, not employing video recording, of Precthl's method on the qualitative assessment of general movements (GMs) in preterm, term and young infants at neurological risk. MATERIALS AND METHODS: One-hundred and fifteen infants consecutively enrolled in our follow-up program were selected for the study (103 preterm and 12 term infants). While being video recorded, each infant's spontaneous motor activity was directly observed and documented using a written proforma. An evaluation of the video was later performed by a different assessor blind to the infant's clinical history. RESULTS: The correlation between the two techniques was significant both at writhing age (birth to 6 weeks post-term age) and at fidgety age (9-15 weeks post-term age). Both methods showed a very high sensitivity for the prediction of cerebral palsy, as no false negatives were observed. The direct assessment showed a lower specificity, particularly during the writhing period. CONCLUSIONS: These results support the use of the direct assessment of GMs when the full application of the standard video observation cannot be routinely applied, restraining the use of video recordings to the abnormal or doubtful cases. This may facilitate the wished integration of the assessment of spontaneous motility into more general protocols of neurological examination and into clinical follow-up programs. PMID- 17428707 TI - Thyroid dysfunction associated with increased low-density lipoprotein cholesterol in epileptic children treated with carbamazepine monotherapy: a causal relationship? AB - OBJECTIVE: Lipid abnormalities and thyroid dysfunction have been reported in patients treated with antiepileptic drugs. The aim of this study was to evaluate prospectively the association between thyroid and lipid profile in children treated with carbamazepine (CBZ) monotherapy. MATERIALS AND METHODS: Thyroid function was evaluated in 18 epileptic children, previously reported with CBZ induced changes in serum lipid profile, before and at 6, 12 and 24 months of CBZ monotherapy. RESULTS: All children had normal thyroid function before the initiation of CBZ treatment. During CBZ therapy thyroid dysfunction, with increased thyrotropin (TSH) and decreased thyroxine (T4), free thyroxine (FT4) and triiodothyronine (T3) was found, while, significant association was revealed between serum low-density lipoprotein cholesterol (LDL-C) and TSH levels at 6 (r=0.469; p=0.043) and 12 (r=0.730; p=0.001) months of treatment. CONCLUSION: Lipid abnormalities may be associated with thyroid hormone disturbance in children treated with CBZ monotherapy. Since thyroid dysfunction and hypercholesterolemia are both associated with a higher atherosclerotic risk special attention and further studies are needed in epileptic patients treated with CBZ monotherapy. PMID- 17428708 TI - Sumatriptan nasal spray in the acute treatment of migraine in adolescents and children. AB - About 4-10% of children and adolescents suffer from migraine. In the last few years, several studies have been performed to assess the efficacy and safety of triptans for the acute treatment of migraine in children and adolescents. Only sumatriptan nasal spray has been approved for the treatment of acute migraine with or without aura in adolescents aged 12-17 years in Europe. This review describes the results of the studies with sumatriptan nasal spray that have been performed in children and adolescents, including a study performed in the Netherlands. PMID- 17428709 TI - Phenytoin desensitization monitored by antigen specific T cell response using carboxyfluorescein succinimidyl ester dilution assay. AB - We evaluated drug-specific T cell responses in a patient with refractory partial seizures and paroxysmal kinesigenic choreoathetosis successfully treated with clinical desensitization to phenytoin. Drug-induced lymphocyte transformation test before desensitization was negative with a stimulation index of 130%. The frequencies and cytokine-producing phenotypes of phenytoin-specific T cells were examined simultaneously by using a carboxyfluorescein succinimidyl ester (CFSE) dilution assay. Before desensitization, the proportion of CFSElow CD4+ cells in whole CD4+ was 3.09%; 13.6% of CFSElow CD4+ cells were stained with anti interferon gamma antibody. After desensitization, phenytoin-specific CFSElow CD4+ cells decreased to background level. These results indicate that CFSE dilution assay will be useful for the diagnosis and monitoring of drug hypersensitivity. PMID- 17428710 TI - Siblings with infantile cerebral stroke and delayed multivessel involvement--a new hereditary vasculopathy? AB - We describe an unusual vasculopathy in two sisters of non-consanguineous parents. The first child developed an acute hemiparesis and focal seizures at the age of 6 months during a febrile illness. Magnetic resonance imaging (MRI) of the brain showed bilateral cortical-subcortical infarction not confined to a vascular territory. Subsequently, the child had a persistent stable neurological deficit. Her younger sister had a similar encephalitis-like episode at the age of 4 months, with left-sided cortical-subcortical ischaemic lesions. Two months later she had left-sided focal seizures. MRI showed a right-sided cortical enhancement, magnetic resonance angiography (MRA) was normal. The neurological deficit was stable and she was seizure free. These episodes were initially interpreted as metabolic strokes, but work-up was normal and mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) was excluded. In their teens both sisters were diagnosed with pulmonary and systemic hypertension and, due to the arterial hypertension, myocardial hypertrophy. Renal artery stenosis, pathological pulmonary arteries, and stenosis and rarefication of coronary arteries were found; the aorta and retinal vessels were normal. Repeat cranial MRI and MRA showed multiple collaterals, while the carotid and basilar arteries were extremely narrowed (moyamoya appearance). We suggest the diagnosis is a hereditary systemic vasculopathy of unknown origin. PMID- 17428711 TI - An optimization method for designing SENSE imaging RF coil arrays. AB - An optimization method in RF coil array design for SENSE imaging is described. Using this method the optimized RF coil geometries can be calculated numerically given the required SENSE imaging performance. Although this method can be applied to optimize the RF coil arrays for both 1D and 2D SENSE imaging, to demonstrate the potential applications of this method, we designed RF coil arrays for 2D SENSE imaging and compared their performance by simulation. An optimized 4 channel receive-only RF coil array designed for 2D SENSE imaging was implemented and tested to demonstrate the feasibility of the proposed technique. Imaging results showed reasonable agreement with the simulations, thus the method can be applied to RF coil array designs for SENSE imaging when optimum imaging performance is desired. PMID- 17428712 TI - Quantitative discrimination of water and hydrocarbons in porous media by magnetization prepared centric-scan SPRITE. AB - MRI has considerable potential as a non-destructive probe of porous media, offering the possibility of rapid quantification of local oil and water content. This potential has not yet, however, been completely realized. In this paper, we explore a general magnetization preparation approach to the discrimination of water and oil in a model, representative, porous medium. These measurements have, as a common element, a centric scan pure phase encode readout based on the SPRITE methodology. Magnetization preparation permits facile T1, T2 and diffusion coefficient mapping as the basis for oil and water discrimination. Diffusion coefficient mapping proved to be the most robust approach to discrimination of oil and water. These methods are illustrated through static experiments and a dynamic immiscible fluid displacement experiment. PMID- 17428713 TI - Sildenafil improves endothelial function in patients with pulmonary hypertension. AB - BACKGROUND: Sildenafil has been shown to be effective in the treatment of pulmonary hypertension, and has favourable effects on endothelial function. Our hypothesis is that a part of the beneficial effects of sildenafil in patients with pulmonary hypertension is due to the improvement of the endothelial function. METHODS: Nine patients (seven females, age 67+/-9 years) with thromboembolic pulmonary hypertension were treated with sildenafil, at a mean dose of 150+/-75 mg/die. At baseline and after 6 months all patients underwent: right-heart catheterization, 6-min walking distance, and a study of endothelial function, including the measure of the flow-mediated vasodilation of the brachial artery, and the dosage of plasma levels of endothelin-1 and von Willebrand factor. RESULTS: During follow-up we found a significant reduction of mean pulmonary artery pressure and arteriolar resistances. Accordingly, the functional capacity improved (an average of+37 m). Sildenafil improved endothelial-dependent vasodilation and reduced plasma concentrations of endothelin-1 (from 4.5+/-0.6 to 3.1+/-0.7 pg/mL; p<0.0001) and von Willebrand factor (from 183.1+/-10.1 to 149.1+/-17.6 mU/mL; p<0.0001). CONCLUSION: Improvement of the endothelial function may represents one of the mechanisms able to explain the favourable effects sildenafil has shown in patients with pulmonary hypertension. PMID- 17428714 TI - Inhaled furosemide does not alleviate respiratory effort during flow-limited exercise in healthy subjects. AB - Expiratory muscle loading results in increased perception of respiratory effort; this response is mediated by non-vagal reflexes originating in the chest wall. Furosemide, due to its vagal effect, might not affect the perception of respiratory effort during expiratory flow-limited incremental exercise. In this study, we compared in nine healthy subjects the following determinants of exercise performance such as respiratory effort (Borg), workload (W'), ventilation (V'E), tidal volume (VT), respiratory frequency (f), and mean inspiratory flow (VT/TI), an index of central respiratory drive, during either standard incremental cycling exercise, or expiratory flow-limited incremental exercise. In addition we examined the effect of inhaled placebo, furosemide (40 or 80 mg) on the perception of respiratory effort following standard incremental cycling exercise and expiratory flow-limited incremental exercise. Compared with standard incremental cycling exercise, expiratory flow-limited incremental exercise increased the Borg score and VT/Ti, and decreased W',V'E ,VT, and f in all subjects at iso-workload. Neither placebo nor furosemide modified peak ventilatory variables, slopes, or intercepts of the relationships of the Borg score with W', V'E, VT/TI and VT during expiratory flow-limited incremental exercise. We conclude that (a) compared with standard incremental exercise, expiratory flow limited exercise increases central respiratory drive and perception of respiratory effort, and (b) furosemide does not affect the sensation of respiratory effort under the present conditions of increased drive to the respiratory muscles. PMID- 17428715 TI - Pigment dispersing hormone generates a circadian response to light in the crayfish, Procambarus clarkii. AB - Photoreceptor cells have been identified as important structures in the organization of the circadian system responsible for the generation and expression of the electroretinogram (ERG) circadian rhythm. They are the structures where the circadian periodicity is expressed (effectors) and which transform information from external light signals to be conducted to the pacemaker in order to induce adjustments of the rhythm (synchronizers). After isolation, eyestalks perfused in a pigment dispersing hormone (PDH) solution, show significant changes in receptor potential (RP) amplitude and duration. Exogenous PDH injected into intact crayfish induces a migration of retinal shielding pigments to a light-adapted state. A single dose of PDH produces advances or delays in the circadian rhythm of response to light of visual photoreceptors. All these effects depend on the circadian phase of PDH application. Consequently, the determination of the action of exogenous PDH on photoreceptor cells proved to be very helpful in understanding some mechanisms underlying the circadian organization of crayfish. PMID- 17428716 TI - Comparative biochemistry and physiology in Latin America over the last decade (1997-2006). AB - We investigate the distribution of Latin American comparative biochemistry and physiology across subject areas and systematic groups. Our study focuses on papers published over the last decade (1997-2006) in four leading topical journals. Brazil dominates the production of papers, followed by Argentina, Mexico and Chile, which together account for 91% of the production in numbers. These countries differ in a number of variables that we discuss in the text. Questions regarding the physiology of wild animals are particularly common, but comparative approaches to study practical issues are also well represented throughout the continent. The most frequent topics in the latter context include the action of snake venoms, the physiology of pathogens and their hosts, and the physiology of domestic or cultured species. The subjects of metabolism, thermoregulation and digestive physiology constitute 40% of the contributions in the database. Regarding systematic groups, most attention has been devoted to vertebrates, mostly anuran amphibians. Other highlighted groups are octodontid rodents, phocid mammals and characid fish. Among invertebrates, malacostracan decapods are by far the best studied group. Many taxa of wild animals are represented by just one or a few studies, thus limited information is available about a large number of Latin American species. PMID- 17428717 TI - Stress biomarkers in rats submitted to swimming and treadmill running exercises. AB - The objective of the present work was to compare stress biomarkers (serum ACTH and corticosterone hormones) during known intensity swimming and treadmill running exercises performed by rats. Adult Wistar rats (n=41) weighing 320-400 g at the beginning and 420-500 g at the end of the experiment, previously adapted to exercise and with Maximal Lactate Steady State (MLSS) already determined were used. The animals were divided into the following subgroups: (1) sacrificed shortly after session of 25 min of exercise (swimming or treadmill) at the MLSS intensity or (2) sacrificed after exhaustive exercise (swimming or treadmill) at intensity 25% higher than MLSS. For comparison, a control group C was sacrificed at rest. Two-way ANOVA was used to identify differences in the stress parameters (P<0.05). At both exercise intensities serum ACTH concentrations were significantly higher for the swimming group compared to running and control groups, while serum corticosterone concentrations in swimming and running groups were significantly higher than in the control group. The differences were more pronounced at the higher intensity (25% higher than MLSS). The swimming group showed higher concentrations for both hormones in relation to the running group. Only acute swimming exercise induced activity of the hypothalamic-pituitary adrenal axis responses expected to stress: elevations in the serum ACTH and corticosterone concentrations. PMID- 17428718 TI - Organic osmolytes in the developing kidney of the Australian brush-tailed possum, Trichosurus vulpecula. AB - Developmental changes in the plasma and urine composition of 120 immature possums, Trichosurus vulpecula, were investigated and correlated with changes in the osmolyte composition of the kidneys. In early life, when the animal is confined in the pouch, the urine is more or less isosmotic with the plasma (300+/ 70 mOsm kg(-1)), but rises steeply in animals more than 90 days old, up to 667+/ 53 mOsm kg(-1) in the oldest age group. In parallel with this change there are corresponding increases in medullary osmolytes. Measured as mmol kg(-1) wet weight, sorbitol was found to increase from 2.1+/-0.8 to 9.7+/-2.2, myo-inositol from 10.9+/-6.2 to 33.6+/-11.7, while the methylamines glycerophosphorylcholine and betaine rose from 6.2+/-0.5 to 15.3+/-3.1 and 3.9+/-2.2 to 9.4+/-2.5 mmol kg( 1) wet weight respectively. Medullary taurine showed no significant changes with age in young possums, while urea increased from 11+/-4.6 to 49+/-7.1 mmol kg(-1) wet weight. These values are similar to, but less than, those found in adult kidneys, but probably further refinement of the concentrating ability of the kidney occurs after the young animal becomes independent. PMID- 17428719 TI - Cathepsin D from Atlantic cod (Gadus morhua L.) liver. Isolation and comparative studies. AB - The isolated cathepsin D-like enzyme from Atlantic cod (Gadus morhua L.) liver was shown to be a monomer with a molecular mass of approximately 40 kDa. It was inhibited by Pepstatin A and had an optimum for degradation of haemoglobin at pH 3.0. The purified enzyme had lower temperature stability than bovine cathepsin D. Antibodies raised against the purified enzyme and against two C-terminal peptides of cod cathepsin D recognized a 40 kDa protein in immunoblotting of the samples from the purification process. Both antisera showed cross reactivity with a similar sized protein in liver from cod, saithe (Pollachius virens L.), Atlantic herring (Clupea harengus L.) and Atlantic salmon (Salmo salar L.). A protein of same size was detected in wolffish (Anarhichas lupus L.) liver with the antibody directed against the purified enzyme. This antibody also recognized the native enzyme and detected the presence of cathepsin D in muscle of cod, saithe, herring and salmon. These antibodies may be useful in understanding the mechanisms of post mortem muscle degradation in fish by comparing immunohistochemical localization and enzyme activity, in particular in cod with different rate of muscle degradation. They may also be used for comparing muscle degradation in different fish species. PMID- 17428720 TI - Ulnar neuropathy type-III at the hand due to chronic non-specific synovitis. PMID- 17428721 TI - Effect of beta-blockers on bone mass and biomechanical parameters of the femoral neck in males with acute myocardial infarction. AB - OBJECTIVES: Evaluate the effect of cardioselective beta-blockers on bone mass and biomechanical properties of the femoral neck in males with acute myocardial infarction. METHODS: Forty males with acute myocardial infarction were studied during one year. Seventy-five percent of the patients (30 patients) were treated with cardioselective beta-blockers and 10 were not similarly treated. A hip densitometry was performed upon release and one year later. The BMD was measured in the femoral neck and in biomechanical elements obtained by DXA. RESULTS: Both groups had similar clinical conditions at the beginning of the study and after a one-year follow-up. No differences in the BMD (0.934+/-0.12 vs. 0.921+/-0.14) were observed in the group without beta-blockers or in the group with beta blockers (0.980+/-0.12 vs. 0.977+/-0.12). No differences were observed in the measured structural parameters. CONCLUSION: The cardioselective beta-blockers do not modify bone mass or the structural bone parameters in males with acute myocardial infarction. PMID- 17428722 TI - The Hsp40 proteins of Plasmodium falciparum and other apicomplexa: regulating chaperone power in the parasite and the host. AB - Extensive structural and functional remodelling of Plasmodium falciparum (malaria)-infected erythrocytes follows the export of a range of proteins of parasite origin (exportome) across the parasitophorous vacuole into the host erythrocyte. The genome of P. falciparum encodes a diverse chaperone complement including at least 43 members of the heat shock protein 40kDa (Hsp40) family, and six members of the heat shock protein 70kDa (Hsp70) family. Nearly half of the Hsp40 proteins of P. falciparum are predicted to contain a PEXEL/HT (Plasmodium export element/host targeting signal) sequence motif, and hence are likely to be part of the exportome. In this review we critically evaluate the classification, sequence similarity and clustering, and possible interactors of the P. falciparum Hsp40 chaperone machinery. In addition to the types I, II and III Hsp40 proteins all exhibiting the signature J-domain, the P. falciparum genome also encodes a number of specialized Hsp40 proteins with a J-like domain, which we have categorized as type IV Hsp40 proteins. Analysis of the potential P. falciparum Hsp40 protein interaction network revealed connections predominantly with cytoskeletal and membrane proteins, transcriptional machinery, DNA repair and replication machinery, translational machinery, the proteasome and proteolytic enzymes, and enzymes involved in cellular physiology. Comparison of the Hsp40 proteins of P. falciparum to those of other apicomplexa reveals that most of the proteins (especially the PEXEL/HT-containing proteins) are unique to P. falciparum. Furthermore, very few of the P. falciparum Hsp40 proteins have human homologs, except for those proteins implicated in fundamental biological processes. Our analysis suggests that P. falciparum has evolved an expanded and specialized Hsp40 protein machinery to enable it successfully to invade and remodel the human erythrocyte, and we propose a model in which these proteins are involved in chaperone-mediated translocation, folding, assembly and regulation of parasite and host proteins. PMID- 17428723 TI - Cytogenetic monitoring by use of the micronucleus assay among hospital workers exposed to low doses of ionizing radiation. AB - The aim of this study was to assess occupationally induced chromosomal damage in a large population of hospital workers exposed to low doses of ionizing radiation. We used the cytokinesis-block micronucleus (CBMN) assay in the peripheral lymphocytes of 132 exposed workers compared with 69 controls matched for gender, age and smoking habits. The CBMN assay was combined with fluorescence in situ hybridization with a human pan-centromeric DNA probe in 32 exposed subjects and 30 controls randomly chosen from the initial populations. Occupational dosimetry records were collected over the last 10-year period and revealed very low exposure levels. The average binucleated micronucleated cell rate (BMCR) was significantly higher in the exposed subjects than in the controls (14.9 per thousand+/-8.1 versus 11.8 per thousand+/-6.5; P=0.011). About one third of the micronuclei were centromere-negative in the exposed and control groups. BMCR significantly positively correlated with donor age in the exposed population; this correlation was at the border of significance in the control group. In the two groups, BMCR was significantly greater in females than in males, and the significant correlation between age and BMCR was observed in the female population, but not in the male one. No effect of smoking habits emerged. Univariate analysis revealed a possible influence of familial cancer history and diagnostic medical radiation dose (estimated from examinations reported in the questionnaire) on BMCR. Multiple regression analysis, taking into account all the previous confounding factors, showed that only occupational exposure status, gender and age had a significant effect on BMCR. In conclusion, the present study shows that chromosomal damage leading to micronucleated lymphocytes is more frequent in hospital workers exposed to ionizing radiation than in controls, despite the very low levels of exposure. PMID- 17428724 TI - GST genotypes and lung cancer susceptibility in Asian populations with indoor air pollution exposures: a meta-analysis. AB - About half of the world's population is exposed to smoke from heating or cooking with coal, wood, or biomass. These exposures, and fumes from cooking oil use, have been associated with increased lung cancer risk. Glutathione S-transferases play an important role in the detoxification of a wide range of human carcinogens in these exposures. Functional polymorphisms have been identified in the GSTM1, GSTT1, and GSTP1 genes, which may alter the risk of lung cancer among individuals exposed to coal, wood, and biomass smoke, and cooking oil fumes. We performed a meta-analysis of 6 published studies (912 cases; 1063 controls) from regions in Asia where indoor air pollution makes a substantial contribution to lung cancer risk, and evaluated the association between the GSTM1 null, GSTT1 null, and GSTP1 105Val polymorphisms and lung cancer risk. Using a random effects model, we found that carriers of the GSTM1 null genotype had a borderline significant increased lung cancer risk (odds ratio (OR), 1.31; 95% confidence interval (CI), 0.95-1.79; p=0.10), which was particularly evident in the summary risk estimate for the four studies carried out in regions of Asia that use coal for heating and cooking (OR, 1.64; 95% CI, 1.25-2.14; p=0.0003). The GSTT1 null genotype was also associated with an increased lung cancer risk (OR, 1.49; 95% CI, 1.17-1.89; p=0.001), but no association was observed for the GSTP1 105Val allele. Previous meta- and pooled analyses suggest at most a small association between the GSTM1 null genotype and lung cancer risk in populations where the vast majority of lung cancer is attributed to tobacco, and where indoor air pollution from domestic heating and cooking is much less than in developing Asian countries. Our results suggest that the GSTM1 null genotype may be associated with a more substantial risk of lung cancer in populations with coal exposure. PMID- 17428725 TI - Molecular identification of the etiological agent of the human anisakiasis in Japan. AB - Anisakis simplex complex presently comprises three sibling species, A. simplex sensu stricto, A. pegreffii and A. simplex C. A. simplex is a common parasite in fishes and cephalopods and capable of causing anisakiasis in humans. Therefore, identification of sibling species of A. simplex was important for human health. In this study, one hundred Anisakis type I larvae isolated from eighty five patients with anisakiasis in Hokkaido and Kyushu in Japan were analyzed by adapting the new molecular method that can identify the sibling species of A. simplex complex. Based on the restriction fragment length polymorphism (RFLP) pattern of ITS regions including 5.8 subunit rRNA gene, we identified two sibling species, A. simplex s. str. and A. pegreffii. However, the infection rate of A. simplex s. str. was significantly higher than that of A. pegreffii. Eighty four (98.8%) out of the eighty five patients were infected with A. simplex s. str. On the contrary, one patients (1.2%) in Kyushu infected with A. pegreffii. This study provided basic information about human infection with A. simplex complex. Furthermore, we suggested that A. simplex s. str. is the most important etiological agent in Japan. PMID- 17428726 TI - Optical absorption and EPR studies on beaverite mineral. AB - The behaviour of transition metal ions in beaverite mineral has been studied by spectroscopic techniques such as electron paramagnetic resonance and absorption spectroscopy in the UV-vis and NIR regions. The ground state of Cu(II) ion in beaverite is confirmed as (2)B(1g) since g(parallel)>g(perpendicular) (2.42>2.097). A resonance noticed at g=2.017 is ascribed to Fe(III) impurity. Two sets of three characteristic bands observed in the optical absorption spectra are assigned to the same transitions, (2)B(1g)-->(2)A(1g), (2)B(1g)-->(2)B(2) and (2)B(1g)-->(2)E(g) of Cu(II) ion in tetragonal field. The presence of Fe(III) bands is supportive evidence for iron impurity in the mineral. Mid infrared spectrum is due to overtones and combination tones of water and hydroxyl groups. PMID- 17428727 TI - Comparative lipidomics analysis of cellular development and apoptosis in two Taxus cell lines. AB - A comparative lipidomics approach was employed to investigate the changes in membrane phospholipids during the procession of cellular development and apoptosis of two plant cell lines, Taxus cuspidata and Taxus chinensis var. mairei. Analysis of lipids by LC/ESI/MS(n) showed more than 90 phospholipid molecular species and indicated significant differences in the abundance throughout a 3-week period. Phosphatidic acid (PA), phosphatidylcholine (PC) and lysophosphatidylcholine (LysoPC) were three important lipid groups that were responsible for the discrimination between the apoptotic T. chinensis var. mairei and living T. cuspidata cells. Continuous increase of phospholipase D (PLD) activity led to PA production in apoptotic T. chinensis var. mairei cells suggesting that the PLD activation and PA formation mediated the apoptosis. Comparison of the profiles of phosphatidylbutanol (PtdBut) with those of PC or phosphatidylethanolamine (PE) indicated that PC rather than PE was the major substrate of PLD in vivo. These results suggest that the alternation of membrane phospholipids may regulate apoptosis, triggering an increase in taxol production of T. chinensis var. mairei cells. PMID- 17428728 TI - Evidence for oxylipin synthesis and induction of a new polyunsaturated fatty acid hydroxylase activity in Chondrus crispus in response to methyljasmonate. AB - Signaling cascades involving oxygenated derivatives (oxylipins) of polyunsaturated fatty acids (PUFAs) are known to operate in response to external stimuli. The marine red alga Chondrus crispus uses both oxygenated derivatives of C18 (octadecanoids) and C20 (eicosanoids) PUFAs as developmental or defense hormones. The present study demonstrates that methyljasmonate (MeJA) triggers a cascade of oxidation of PUFAs leading to the synthesis of prostaglandins and other oxygenated fatty acids. As a result of a lipoxygenase-like activation, MeJA induces a concomitant accumulation of 13-hydroxy-9Z,11E-octadecadienoic acid (13 HODE) and 13-oxo-9Z,11E-octadecadienoic acid (13-oxo-ODE) in a dose-dependent manner in C. crispus. Furthermore, MeJA increases the level of mRNA encoding a gluthatione S-transferase and induces the activity of a new enzyme catalyzing the regio- and stereoselective bisallylic hydroxylation of polyunsaturated fatty acids from C(18) to C(22). The enzyme selectively oxidized the omega minus 7 carbon position (omega-7) and generated the stereoselective (R)-hydroxylated metabolites with a large enantiomeric excess. The enzyme specificity for the fatty acid recognition was not dependent of the position of double bonds but at least requires a methylene interrupted double bond 1,4-pentadiene motif involving the omega-7 carbon. PMID- 17428729 TI - The lipid-binding SEC14 domain. AB - Protein-lipid interactions are important for protein targeting, signal transduction, lipid transport, lipid biosynthesis, lipid metabolism, and the maintenance of cellular compartments and membranes. Specific lipid-binding protein domains, such as PH, FYVE, PX, PHD, C2 and SEC14 homology domains, mediate interactions between proteins and specific phospholipids. Here we review the published literature, plus some of our most recent unpublished findings, regarding the biology of the SEC14 domain, also known as CRAL_TRIO domain. PMID- 17428730 TI - Safety issues and prospects for future generations of PPAR modulators. AB - Because of their wide range of actions on glucose homeostasis, lipid metabolism and vascular inflammation, peroxisome proliferator-activated receptors (PPARs) are promising targets for the development of new drugs for the treatment of metabolic disorders such as diabetes, dyslipidemia and atherosclerosis. In clinical practice, PPARalpha agonists, such as the already available fibrates, improve dyslipidemia, while PPARgamma agonists, such as thiazolidinediones, improve insulin resistance and diabetes. The complementary action of simultaneous activation of each PPAR in patients suffering from metabolic syndrome and type 2 diabetes has led to new pharmacological strategies focused on the development of agonists targeting more than one receptor such as the dual PPARalpha/gamma agonists. However, despite the proven benefits of targeting PPARs, safety concerns have recently led to late stage development failures of various PPAR agonists including novel specific PPARgamma agonists and dual PPARalpha/gamma agonists. These safety concerns include potential carcinogenicity in rodents, signs of myopathy and rhabdomyolysis, increase in plasma creatinine and homocysteine, weight gain, fluid retention, peripheral edema and potential increased risk of cardiac failure. Although the discontinued compounds shared common side effects, the reason for discontinuation was always compound specific and the toxicological or adverse effects which have motivated the discontinuation could be either due to the activation of PPARgamma, PPARalpha or both (class effect) or due to a PPAR unrelated effect. Thus, the risk evaluation of each adverse effect should be viewed on a case by case basis considering both the PPAR profile of the drug, its absorption/distribution profile, the nature of the side effect and the putative PPAR-related mechanism of action. This review mainly focuses on the preclinical and clinical adverse events of PPAR agonists that could be of concern when considering the development of new PPAR agonists. The selective modulation of PPAR activities is a promising approach to develop new drugs with preserved efficacy but diminished adverse effects. PMID- 17428731 TI - SELEX experiments: new prospects, applications and data analysis in inferring regulatory pathways. AB - Systematic Evolution of Ligands by EXponential enrichment (SELEX) is an experimental procedure that allows extraction, from an initially random pool of oligonucleotides, of the oligomers with a desired binding affinity for a given molecular target. The procedure can be used to infer the strongest binders for a given DNA or RNA binding protein, and the highest affinity binding sequences isolated through SELEX can have numerous research, diagnostic and therapeutic applications. Recently, important new modifications of the SELEX protocol have been proposed. In particular, a modification of the standard SELEX procedure allows generating a dataset from which protein-DNA interaction parameters can be determined with unprecedented accuracy. Another variant of SELEX allows investigating interactions of a protein with nucleic-acid fragments derived from the entire genome of an organism. We review here different SELEX-based methods, with particular emphasis on the experimental design and on the applications aimed at inferring protein-DNA interactions. In addition to the experimental issues, we also review relevant methods of data analysis, as well as theoretical modeling of SELEX. PMID- 17428732 TI - Superior vena cava obstruction by tumour thrombus in invasive thymoma: diagnosis and surgical management. AB - Thymomas are common mediastinal tumours. We report a rare case of thymoma invasion into the superior vena cava with resultant venous obstruction. The tumour was resected. The superior vena cava and left brachiochephalic vein were reconstructed with autologous pericardial patch. PMID- 17428734 TI - An unusual case of late bioprosthetic mitral valve thrombosis successfully managed with anticoagulation. AB - In this report, we present the case of a patient with hemodynamically significant thrombosis of a mitral bioprosthesis occurring 11 years after valve replacement. The diagnosis was suspected on the basis of a subacute increase in the transvalvular gradient seen on transthoracic echocardiography and was con- firmed by transesophageal echocardiography which disclosed unusual features. A hypercoagulable work up showed raised plasma factor VIII levels. The patient was successfully managed with oral anticoagulation. PMID- 17428735 TI - Acetylcholinesterase activity in the earthworm Eisenia andrei at different conditions of carbaryl exposure. AB - Recent reports have stressed the need for a better understanding of earthworm biomarker responses. We aimed at investigating acethylcholinesterase (AChE) activity in the earthworm Eisenia andrei after exposure to carbaryl or its commercial formulation Zoril 5 under different in vitro and in vivo experiments. In addition, lysosome membrane stability was assessed by neutral red retention assay in the same experimental conditions. AChE basal Km and Vm values were about 0.16 mM and 41 nmol min(-1) mg protein(-1), respectively. Carbaryl dose dependently decreased Vmax, while not affecting Km values. Carbaryl reduced earthworm AChE activity within 1 day of in vivo exposure to contaminated filter paper. Tested on soil, carbaryl inhibited AChE with the maximum effect after 3 days; in contrast, lysosome membrane stability of coelomocytes indicated a maximum toxicity after one day, followed by a recovery. AChE inhibition by Zoril 5 was highest after one day, while lysosome membrane stability declined progressively. In all cases, carbaryl dose-dependently decreased Vmax while not affecting Km values. In conclusion, E. andrei AChE activity assessed in vitro is dose-dependently inhibited by the carbamate compound carbaryl, which acts as a pure competitive inhibitor. In vivo experiments suggested that pure and co formulated carbaryl have different time and/or dose dependent effects on earthworms. Our results further support the use of AChE inhibition as an indicator of pesticide contamination, to be included in a battery of biomarkers for monitoring soil toxicity. PMID- 17428736 TI - Lipid dynamics in goldfish (Carassius auratus) during a period of gonadal recrudescence: Effects of beta-sitosterol and 17beta-estradiol exposure. AB - The potential for contaminants to alter lipid or cholesterol dynamics in fish is rarely investigated and may include critical physiological endpoints that are impacted by exposure to endocrine-active substances. The current study investigated plasma and tissue lipid dynamics over a period of recrudescence in goldfish, while also examining the potential for beta-sitosterol (beta-sit), a phytosterol and 17beta-estradiol (E(2)), an endogenous estrogen, to alter lipid homeostasis. Goldfish were exposed to 0 microg/g (no chemical; control), 200 microg/g beta-sit (72.3% sitosterol mixture) or 10 microg/g 17beta-estradiol (E2) via Silastic implants for a period of five months. Plasma lipids peaked in control fish coincident with maximum liver size, while gonadal cholesterol concentration was highest concomitant with maximum gonad size. Plasma lipid concentrations were highly affected by E2 but not beta-sit exposure; E2 elevated total cholesterol (p<0.001) and triglyceride (TG; p<0.001) and decreased high density lipoprotein (HDL) concentration (p<0.001) in male fish. Tissue cholesterol concentrations were minimally affected by beta-sit exposure, while hepatic cholesterol concentrations were increased in E2 exposed females (p=0.041), indicating elevated liver lipogenesis in response to E2, but not beta sit, exposure. The present study demonstrates differential effects by beta-sit and E2 on plasma lipoprotein profile and TG concentration and indicates estrogen specific effects on hepatic lipid metabolism during gonadal development. PMID- 17428737 TI - Clinical recognition of Al type amyloidosis of the luminal gastrointestinal tract. AB - BACKGROUND & AIMS: Amyloidosis is characterized by the pathologic deposition of specific proteins throughout the body. Gastrointestinal involvement with amyloid associated with plasma cell dyscrasias (AL type amyloidosis) is common, but systematic description of the condition is lacking. The aim of this investigation was to characterize the clinical presentation, endoscopic findings, and histopathologic correlates in a series of patients with systemic AL amyloidosis of the luminal gastrointestinal tract. METHODS: Eligible patients were identified by interrogating the histopathology database of our institution during a 14-year time period. Medical record, histopathologic, and laboratory data were collected, analyzed, and correlated with endoscopic findings. RESULTS: Nineteen patients with systemic AL amyloidosis of the luminal gastrointestinal tract were identified. Gastrointestinal symptoms or signs related to amyloid involvement were noted in 95% of patients; abdominal pain, change in bowel habits, overt gastrointestinal bleeding, and complaints related to altered motility were the predominant presentations. Endoscopic abnormalities were found in nearly three fourths of patients, including ulcerations and submucosal hematomas. When gastrointestinal bleeding was the presenting symptom, submucosal hematomas were a common finding during endoscopic evaluation. CONCLUSIONS: AL type amyloidosis of the luminal gastrointestinal tract is a rare disease that presents with common, nonspecific complaints. The endoscopic detection of a submucosal hematoma in the setting of gastrointestinal bleeding in patients with plasma cell dyscrasias should raise suspicion for the disease. PMID- 17428738 TI - Granular cell tumor in colonic polyp found on screening colonoscopy. PMID- 17428739 TI - Chronic hepatitis B virus carriers in the immunotolerant phase of infection: histologic findings and outcome. AB - BACKGROUND & AIMS: The indication for histologic evaluation of the liver is controversial for patients in the immunotolerant phase of chronic hepatitis B virus (HBV) infection. METHODS: Results of liver biopsy examination and follow-up evaluation were assessed retrospectively in 40 patients who satisfied the following criteria: presence of hepatitis B surface antigen and hepatitis B e antigen, HBV DNA level greater than 10(7) cp/mL, normal alanine aminotransferase level, absence of co-infection with other viruses, and absence of antiviral or immunosuppressive treatment. RESULTS: On liver biopsy examination, according to the Metavir scoring system, fibrosis was absent in 20 patients, and mild (F1) in 20 patients. During a median follow-up period of 37.7 months in 31 patients, loss of tolerance was observed at a median age of 30.7 years in 12 (38%): 6 had transition to inactive disease, 3 developed chronic hepatitis, and 3 had a transient increase of alanine aminotransferase levels. Among baseline characteristics, only alanine aminotransferase levels were significantly higher in patients with subsequent loss of tolerance. CONCLUSIONS: In patients in the immunotolerant phase of chronic HBV infection, liver biopsy examination shows only minimal changes and probably is unnecessary. Loss of tolerance, occurring at a median age of 30.7 years, is characterized by a rapid transition to an inactive carrier state in two thirds of patients, and to chronic hepatitis in one third of patients. PMID- 17428740 TI - Proteases in irritable bowel syndrome: a lot more than just digestive enzymes. PMID- 17428741 TI - Primary endpoints for irritable bowel syndrome trials: a review of performance of endpoints. AB - The choice of primary endpoint for a clinical trial is one of the most important determinants of the ability of a clinical trial to demonstrate efficacy of therapeutic agents. Although there are still no clear, universally accepted guidelines on the definition of clinical benefit for irritable bowel syndrome (IBS), consensus guidelines stress the importance of using validated endpoints. This article reviews the evidence available in the literature on the psychometric validation and performance of the 3 endpoints recommended by the Rome III Committee for use as primary endpoints in treatment trials of IBS. The Rome III Committee recommends 2 types of measures: binary endpoints addressing the construct of relief (that is, adequate relief and satisfactory relief) and an integrative symptom questionnaire that addresses the change in severity of a representative group of symptoms of IBS (that is, the IBS Severity Scale). The current evidence suggests that at present, adequate relief should be recognized by regulatory authorities as an acceptable primary endpoint in clinical trials. This analysis also suggests that data from individual clinical trials should be pooled and undergo meta-analysis, and that prospective studies should be considered to further characterize the performance of available endpoints as outcome measures in pharmacotherapeutic trials in IBS. PMID- 17428742 TI - Eosinophilic gastroenteritis. PMID- 17428743 TI - Correlation of duodenal histology with tissue transglutaminase and endomysial antibody levels in pediatric celiac disease. AB - BACKGROUND & AIMS: IgA antibodies against tissue transglutaminase (TTGA) and endomysium (EMA) are sensitive and specific markers for celiac disease (CD). Data correlating TTGA and EMA levels with degree of villous atrophy are limited. We compared duodenal histopathology in pediatric CD patients with TTGA and EMA serologies, symptoms, height, and weight. METHODS: We identified 117 pediatric patients retrospectively who had serologic testing for IgA TTGA and IgA EMA and duodenal biopsies graded by modified Marsh criteria as 0-3c. Data were analyzed with Spearman rank correlation and multinomial logistic regression. RESULTS: IgA TTGA (r = .704, P < .001) and IgA EMA (r = 0.740, P < .001) correlated with intestinal villous atrophy in pediatric CD patients by Spearman rank correlation. Similar correlations were found in a subset of 23 patients younger than 3 years of age. Multinomial logistic regression revealed increased probability of Marsh 3a or greater changes with increasing TTGA or EMA levels. Strongly positive antibody levels (TTGA >100 units or EMA titer >1:1280) were highly specific (>98%) for Marsh 3a or greater lesions. Among symptoms, abdominal distention and diarrhea were associated with abnormal histology. CONCLUSIONS: IgA TTGA and EMA levels correlate with duodenal villous atrophy in pediatric CD patients. IgA TTGA >100 or EMA >1:1280 were nearly always associated with CD histopathology. With further validation of this observation, strongly positive titers might be considered sufficient for diagnosis of pediatric patients at risk for CD. Symptoms, height, and weight are not reliable predictors of CD. PMID- 17428744 TI - Alternative designs of phase II trials considering response and toxicity. AB - Phase II clinical trials in oncology are used to initially evaluate the therapeutic efficacy of a new treatment regimen. Simon's two-stage design is commonly used for such trials. However, he only focused on the "response rate", the proportion of patients experiencing tumor regression. In clinical practice, it is preferred of a sequential design to monitor antitumor activity as well as toxicity. Conaway and Petroni proposed a method for designing phase II trials on the basis of both treatment efficacy and safety, which imply an equal weighing of response and toxicity. In this paper, we developed an alternative test to cope with the trade-off between safety and efficacy. The main idea is to control for the marginal type I errors of response rate and toxicity rate separately. We provide guides on searching the stopping and rejecting regions and determination of sample size. The proposed method has advantage over other designs, including those of Conaway and Petroni's and Bryant and Day's, that it can definitely control one type I error of the interests such as treatment antitumor activity or safety and is robust against the real association parameter. Furthermore, it is conceptive intuitive, very simple to implement, and also feasible for the requirement of small sample size in a phase II trial. PMID- 17428745 TI - Evidence for non-chemical, non-electrical intercellular signaling in intestinal epithelial cells. AB - Synchrony between mechanically separated biological systems is well known. We posed the question: can cells induce synchronous behavior in neighboring cells which are mechanically separated and which cannot communicate via chemical or electrical mechanisms. Caco-2 cell cultures were divided into three groups. "Inducer" cells were exposed to H(2)O(2). "Detector" cells were placed in separate containers near the inducer cells but were not exposed to H(2)O(2). Control cells were exposed to fresh media and were kept in a distant laboratory area. Samples were measured for total protein concentration, NFkappaB activation and structural changes, 10, 30 and 60 min after exposure respectively. Exposing inducer cells to H(2)O(2) resulted in a significant reduction in total protein content (-50%), an increase in nuclear NFkappaB activation (+38%), and structural damage (56%) compared to controls. There was a similar reduction in total protein content (-48%), increase in the nuclear fraction of NFkappaB (+35%) and structural damage (25%) in detector cells. These findings provide evidence in support of a non-chemical, non-electrical communication. This signaling system possibly plays a role in synchronous, stimulus-appropriate cell responses to noxious stimuli and may explain a number of cellular behaviors that are hard to explain based only on conventional cell signaling systems. PMID- 17428746 TI - Effect of shape on the dielectric properties of biological cell suspensions. AB - In this note, we analyze the effect of cell shape on the dielectric and conductometric behavior of biological cell suspension, in a frequency range where the interfacial polarization characteristic of highly heterogeneous systems occurs. We consider two different families of curves, both of them capable of generating a variety of symmetric or asymmetric shapes, ranging from oval, to dog bone like, to lemniscate curves. These curves, which differ from those generally employed in dielectric models of biological cell suspensions, describe in principle different cells including discocytes, cup-shaped cells, pear-shaped cells, dumbbells and cells with spherical protrusions or invaginations. Our analysis, based on a numerical solution of the Laplace equation by means of boundary element methods, is carried out in the attempt of separating the contributions associated with the different electrical properties of the dielectric media involved from the ones mainly associated with the shape of the cell. We determine the dielectric strength of the dielectric dispersion for a variety of cell shapes and the phenomenological correlation between this parameter of the relaxation and the cell geometry is briefly discussed and commented. PMID- 17428747 TI - Monitoring ATM kinase activity in living cells. AB - DNA double strand breaks (DSB) in mammalian cells result in the activation of the ATM protein kinase. This leads to phosphorylation of numerous downstream transducer and effector proteins that coordinate a cellular response including DNA repair, cell cycle arrest or apoptosis. We have developed a reporter protein that allows the measurement of ATM kinase activity in single living cells. This CFP-YFP FRET-based biosensor uses an ATM phosphorylation site and an FHA phosphospecific binding domain to produce a phosphorylation-induced change in conformation, which alters the FRET efficiency between CFP and YFP. We show that the reporter provides a measurable output in response to DSBs and is specific for ATM over ATR or DNA-PK. We expect the description of the spatiotemporal dynamics of ATM activity in living cells that this reporter provides will be helpful in providing a more detailed understanding of the DNA damage response. PMID- 17428748 TI - Postnatal development of GABAB-receptor-mediated modulation of potassium currents in brainstem respiratory network of mouse. AB - The GABA(B)-receptor is known to activate a potassium conductance that is inwardly-rectifying, Ba(2+)-sensitive and mediated by G-protein-coupled mechanism. The network that generates respiratory rhythm is located in the brainstem and is modulated by GABA(B)-receptors. The present study investigated the mechanisms by which GABA(B)-receptor activation modulates respiratory rhythm and how these effects change during the first 2 weeks of postnatal development (P0-P15). Whole-cell patch clamp recordings were obtained from inspiratory neurons in the ventral respiratory column of acute brain stem slice of mouse. In presence of TTX and cadmium, application of baclofen, a GABA(B)-receptor agonist, activated an inwardly-rectifying potassium current. The reversal potential of the current was around -78 mV, which was close to the calculated equilibrium potential of potassium. The action of baclofen was dose-dependent and could be partially blocked (>85%) by a selective GABA(B)-receptor antagonist CGP 55845A. The current density of the baclofen-activated potassium currents increased over the first 2 postnatal weeks. At the cellular level, baclofen-activated potassium currents hyperpolarized inspiratory neurons in a concentration- and age-dependent manner. At the network level, the frequency of the respiratory rhythm decreased or was abolished depending on the concentration of baclofen applied. Our results indicate that the endogenous modulation of respiratory rhythm by GABA(B) receptors that we have demonstrated previously is mediated at least in part through activation of an inwardly rectifying K(+) conductance and that this effect increases postnatally. PMID- 17428749 TI - Low molecular weight protein tyrosine phosphatase (LMW-PTP) and its possible physiological functions of redox signaling in the eye lens. AB - Low molecular weight protein tyrosine phosphatase (LMW-PTP) was cloned from human lens epithelial B3 cells (HLE B3) and the recombinant enzyme was purified to homogeneity. The pure enzyme reacted positively with anti-LMW-PTP antibody, displayed tyrosine-specific phosphatase activity and was extremely sensitive to H(2)O(2). The inactivated LMW-PTP could be regenerated by thioltransferase (TTase)/GSH system as demonstrated by both activity assay and by mass spectrometry (MS). The MS study also showed that an intramolecular disulfide bond was formed between C13 and C18 at the active site, and was reduced by the TTase/GSH system. The putative role of LMW-PTP in regulating platelet derived growth factor (PDGF)-stimulated cell signaling was demonstrated in wild type mouse lens epithelial cells (LEC) in which LMW-PTP was transiently inactivated, corroborated with the transient phosphorylation of Tyr857 at the active site of PDGF receptor and the downstream signaling components of Akt and ERK1/2. In contrast, LMW-PTP activity in PDGF-stimulated LEC from TTase(-/-) mice was progressively lost, concomitant with the high basal and sustained high phosphorylation levels at Tyr857, Akt and ERK1/2. We conclude that the reversible LMW-PTP activity regulated by ROS-mediated oxidation and TTase/GSH reduction is the likely mechanism of redox signaling in lens epithelial cells. PMID- 17428750 TI - The influence of local and systemic preconditioning on oxygenation, metabolism and survival in critically ischaemic skin flaps in pigs. AB - Stress proteins represent a group of highly conserved intracellular proteins that provide adaptation against cellular stress. The present study aims to elucidate the stress protein-mediated effects of local hyperthermia and systemic administration of monophosphoryl lipid A (MPL) on oxygenation, metabolism and survival in bilateral porcine random pattern buttock flaps. Preconditioning was achieved 24h prior to surgery by applying a heating blanket on the operative site (n = 5), by intravenous administration of MPL at a dosage of 35 microg/kg body weight (n = 5) or by combining the two (n = 5). The flaps were monitored with laser Doppler flowmetry, polarographic microprobes and microdialysis until 5h postoperatively. Semiquantitative immunohistochemistry was performed for heat shock protein 70 (HSP70), heat shock protein 32 (also termed haem oxygenase-1, HO 1), and inducible nitrc oxide synthase (iNOS). The administration of MPL increased the impaired microcirculatory blood flow in the proximal part of the flap and partial oxygen tension in the the distal part by approximately 100% each (both P<0.05), whereas both variables remained virtually unaffected by local heat preconditioning. Lactate/pyruvate (L/P) ratio and glycerol concentration (representing cell membrane disintegration) in the distal part of the flap gradually increased to values of approximately 500 mmol/l and approximately 350 micromol/l, respectively (both P<0.01), which was substantially attenuated by heat application (P<0.01 for L/P ratio and P<0.05 for glycerol) and combined preconditioning (P<0.01 for both variables), whereas the effect of MPL was less marked (not significant). Flap survival was increased from 56% (untreated animals) to 65% after MPL (not significant), 71% after heat application (P<0.05) and 78% after both methods of preconditioning (P<0.01). iNOS and HO-1 were upregulated after each method of preconditioning (P<0.05), whereas augmented HSP70 staining was only observed after heat application (P<0.05). We conclude that local hyperthermia is more effective in preventing flap necrosis than systemic MPL administration because of enhancing the cellular tolerance to hypoxic stress, which is possibly mediated by HSP70, whereas some benefit may be obtained with MPL due to iNOS and HO-1-mediated improvement in tissue oxygenation. PMID- 17428751 TI - Comment on 'Reconstruction of pectus excavatum with silicone implants'. PMID- 17428752 TI - Double-pedicle abdominal perforator free flaps for unilateral breast reconstruction: new horizons in microsurgical tissue transfer to the breast. AB - The DIEAP (deep inferior epigastric artery perforator) flap is a suitable option for breast reconstruction resulting in excellent aesthetic outcome, and minimal donor site morbidity. Contraindications for use of the DIEAP flap may include previous abdominal liposuction and/or surgery, or lack of abdominal tissue. The purpose of this paper is to describe options of using abdominal perforator flaps, based on double-pedicle techniques, despite these contraindications. A retrospective evaluation was carried out on a series of 16 patients who required abdominal double-pedicle free perforator flaps for unilateral breast reconstruction since June 2002. The indications were multiple abdominal scars, previous abdominal liposuction and thin patients in five, three and eight cases, respectively. Preoperative mapping of the vascular network was done using Duplex and/or multi-detector CT scan imaging. Clinical evaluation of medical charts was done regarding patients' characteristics, surgical techniques, ischaemia/total operative time and complications. A clinical evaluation was done on all patients with average follow up of 15 months. Fat necrosis was investigated clinically and by mammogram examination. Different microsurgical techniques were performed to provide enough blood supply to the requested flaps: Perforator (P) to contralateral Deep Inferior Epigastric (DIE) anastomosis (P/DIEAP), in two patients; bilateral DIE vessels (DIEAP/DIEAP) in seven patients; and DIE with SIE (superficial inferior epigastric) vessels in seven patients (DIEAP/SIEA). One pedicle was always anastomosed to the internal mammary vessels. The second pedicle was anastomosed end-to-end to a side branch of the DIE or end-to-side with the DIE pedicle in 13 cases. The thoracodorsal vessels were used as recipient vessels for the second pedicle in three cases. Average operative time was 6h 30min (range 5h 30min-8h). All 16 flaps survived and fat necrosis occurred in one case. The harvesting of perforator free flaps may be contraindicated in some patients, however they are still a feasible option as long as the vessels to the skin are present. Preoperative planning combined with high expertise in microsurgical techniques are the key points in the high success rate in these difficult cases. PMID- 17428754 TI - Comparison of dynamic (effortful) touch by hand and foot. AB - Spatial perception by dynamic touch is a well-documented capability of the hand and arm. Morphological and physiological characteristics of the foot and leg suggest that such a capability may not generalize to that putatively less dexterous limb. The authors examined length perception by dynamic touch in a task in which weighted aluminum rods were grasped by the hand and wielded about the wrist or secured to the foot and wielded about the ankle. Participants' (N = 10) upper and lower extremities were comparable in terms of (a) the accuracy and consistency of length perception and (b) their sensitivity to manipulations of the moments of the mass distribution of the rods. The authors discuss those results in terms of the field-like structure of the haptic perceptual system, an organization that may underlie what appears to be functional, rather than anatomical, specificity. PMID- 17428755 TI - Evidence of proactive forefoot control during landings on inclined surfaces. AB - The notion of proactive control of landings is generally accepted, and some underlying mechanisms have already been described. Only little is known on adjustments at the foot level, however. The authors therefore investigated the foot and ankle behavior of 24 participants as they landed on differently inclined surfaces. A 4-segment model of the foot and ankle provided 3-dimensional kinematics. They also analyzed activation patterns from several muscles and the ground reaction force. Participants anticipated the different surfaces, as shown by the forefoot kinematics and the activation patterns before touch down. Anticipation of the surface inclination led to adjustments in forefoot orientation and probably also in joint stiffness. The authors suggest that those adjustments tend to enhance the self-stabilizing potential of the mechanical system. The enhancement of that potential would ease the subsequent stabilization, reducing the demands on the neural system. PMID- 17428756 TI - Planning and control of sequential rapid aiming in adults with Parkinson's disease. AB - Eight people with Parkinson's disease (PD), 8 age-matched older adults, and 8 young adults executed 3-dimensional rapid aiming movements to 1, 3, 5, and 7 targets. Reaction time, flight time, and time after peak velocity to the 1st target indicated that both neurologically healthy groups implemented a plan on the basis of anticipation of upcoming targets, whereas the PD group did not. One suggested reason for the PD group's deficiency in anticipatory control is the greater variability in their initial force impulse. Although the PD group scaled peak velocity and time to peak velocity similarly to the other groups, their coefficients of variation were greater, making consistent prediction of the movement outcome difficult and thus making it less advantageous to plan too far in advance. A 2nd finding was that the PD group exhibited increased slowing in time after peak velocity in the final segments of the longest sequence, whereas the other 2 groups did not. The increased slowing could be the result of a different movement strategy, increased difficulty modulating the agonist and antagonist muscle groups later in the sequence, or both. The authors conclude that people with PD use more segmented planning and control strategies than do neurologically healthy older and young adults when executing movement sequences and that the locus of increased bradykinesia in longer sequences is in the deceleration phase of movement. PMID- 17428757 TI - Symmetry constraints mediate the learning and transfer of bimanual coordination patterns across planes of motion. AB - The authors investigated whether neuromuscular and directional constraints are dissociable limitations that affect learning and transfer of a bimanual coordination pattern. Participants (N = 9) practiced a 45 degrees muscular relative phasing pattern in the transverse plane over 4 days. The corresponding to-be-learned spatial relative phasing was 225 degrees. Before, during, and following practice, the authors administered probe tests in the sagittal plane to assess transfer of learning. In the probe tests, participants performed various patterns characterized by different muscular and spatial relative phasing (45 degrees, 45 degrees, 45 degrees, 225 degrees, 225 degrees, 45 degrees, and 225 degrees, 225 degrees). The acquisition of the to-be-learned pattern in the transverse plane resulted in spontaneous positive transfer of learning only to coordination patterns having 45 degrees of spatial relative phase, irrespective of muscular phasing. Moreover, transfer occurred in the sagittal plane to coordination patterns that had symmetry properties similar to those of the to-be learned pattern. The authors conclude that learning and transfer of spatial features of coordination patterns from the transverse to the sagittal plane of motion are mediated by mirror-symmetry constraints. PMID- 17428758 TI - Postural stabilization of perceptual but not cognitive performance. AB - In 2 experiments, the authors independently varied the degree of cognitive and perceptual difficulty of suprapostural tasks. Participants were 23 students in Experiment 1 and 15 in Experiment 2. Increases in perceptual difficulty tended to be correlated with decreases in the variability of postural sway, consistent with the hypothesized functional integration of postural control with suprapostural tasks. Sway variability was not influenced by changes in the cognitive difficulty of tasks when perceptual difficulty was held constant, contrary to predictions derived from the perspective that postural and suprapostural activities compete for a limited pool of central processing resources. The results underscore the need for researchers to differentiate between suprapostural tasks that require perceptual contact with the environment and those that do not. PMID- 17428759 TI - Developmental differences in the use of visual information during a continuous bimanual coordination task. AB - The authors examined the influence of different amounts of visual information when children 4 (CH4), 6 (CH6), and 8 (CH8) years of age, and adults (n = 12 in each group) performed a steady-state bimanual circle-drawing coordination task at self-selected speeds. All participants maintained in-phase coordination, but different strategies for maintaining the pattern emerged. A predictable relationship between variability and age was not observed, in that the CH8 group was not necessarily more consistent than the CH6 and CH4 groups. The authors conclude that children are transitioning from dependence on kinesthetic feedback to reliance on visual feedback around age 8, as suggested by L. Hay, C. Bard, M. Fleury, and N. Teasdale (1991; L. Hay, M. Fleury, C. Bard, & N. Teasdale, 1994; L. Hay & C. Redon, 1997), and that future studies are needed to further explore visual and kinesthetic feedback as potential control parameters during coordination tasks in developing children. PMID- 17428761 TI - Unconventional nanotubes self-assembled in alumina channels: morphology and surface potential of isolated nanostructures at surfaces. AB - Synthetic nanographenes have been self-assembled from solution on the surface of nanometric channels of an alumina membrane template. By controlling the interplay between intermolecular and interfacial interactions, the molecules have been adsorbed either 'face-on' or 'edge-on' on the pore's surfaces, leading to the formation of columnar stacks in the latter case. Upon thermal treatment at high temperature, the molecular cross-linking of the columns has been triggered, transforming the delicate supramolecular arrangement into robust carbon nanotubes, with the graphitic planes at predetermined orientations with respect to the tube axis. Scanning force microscopy characterization of single nanotubes deposited from suspensions on mica showed that the nanotubes can self-assemble on flat surfaces adopting preferential alignments which reflect the threefold symmetry of the mica substrate. Kelvin probe force microscopy studies revealed that the nanotubes possess a surface potential much smaller than the work function of both graphite and conventional vacuum-processed nanotubes, providing evidence for their more confined electronic structure. PMID- 17428762 TI - Energy migration in conjugated polymers: the role of molecular structure. AB - Conjugated polymers undergo facile exciton diffusion. Different molecular structures were examined to study the role of the excited state lifetimes and molecular conformations on energy transfer. There is a clear indication that extended fluorescence lifetimes give enhanced exciton diffusion as determined by fluorescence depolarization measurements. These results are consistent with a strong electronic coupling or Dexter-type energy transfer as the dominating mechanism. The control of polymer conformations in liquid crystal solvents was also examined and it was determined that more planar conformations gave enhanced energy transfer to emissive low band-gap endgroups. PMID- 17428763 TI - Soft materials with graphitic nanostructures. AB - This review article focuses on our recent studies on novel soft materials consisting of carbon nanotubes. Single-walled carbon nanotubes, when suspended in imidazolium ion-based ionic liquids and ground in an agate mortar, form physical gels (bucky gels), where heavily entangled bundles of carbon nanotubes are exfoliated to give highly dispersed, much finer bundles. By using bucky gels, the first printable actuators that operate in air for a long time without any external electrolyte are developed. Furthermore, the use of polymerizable ionic liquids as the gelling media results in the formation of electroconductive polymer/nanotube composites with enhanced mechanical properties. The article also highlights a new family of nanotubular graphite, via self-assembly of amphiphilic hexabenzocoronene (HBC) derivatives. The nanotubes consist of a graphitic wall composed of a great number of pi-stacked HBC units and are electroconductive upon oxidation. The use of amphiphilic HBCs with functional groups results in the formation of nanotubes with various interesting properties. PMID- 17428764 TI - The development of self-assembled liquid crystal display alignment layers. AB - From simple pocket calculators to mobile telephones and liquid crystal display (LCD)-TV, over the past few decades, devices based on LCD technology have proliferated and can now be found in all conceivable aspects of everyday life. Although used in cutting-edge technology, it is surprising that a vital part in the construction of such displays, namely the alignment layer, relies essentially on a mechanical rubbing process, invented almost 100 years ago. In this paper efforts to develop alignment layers (also called command layers) by processes other than rubbing, namely self-assembly of molecular and macromolecular components will be discussed. Two topics will be presented: (i) tuneable command layers formed by stepwise assembling of siloxane oligomers and phthalocyanine dyes on indium tin oxide surfaces and (ii) command layers formed by self-assembly of porphyrin trimers. The potential use of these layers in sensor devices will also be mentioned. PMID- 17428765 TI - What determines the mobility of charge carriers in conjugated polymers? AB - In a conjugated polymer, the mobility of charge carriers is not a well-defined coefficient of a particular material as it is in an inorganic crystalline semiconductor but depends on the time domain of detection. On a time-scale of typically 100 fs, the on-chain mobility is ultra-high and controlled by the electronic band width of the polymer chain. When a carrier hits a chain imperfection, subsequent mesoscopic on-chain motion is retarded and controlled by intrachain disorder to which the chain environment contributes. Macroscopic transport commences after a time when interchain carrier jumps become rate limiting. It is routinely probed by time-of-flight experiments and can be rationalized in terms of random walk within a rough energy landscape. Experimental signatures of the various modes of transport are discussed. PMID- 17428766 TI - Expanding benzene to giant graphenes: towards molecular devices. AB - Phenylene-based conjugated materials provide versatile platforms for the development of molecular devices. Architectures of one- and two-dimensional polyphenylenes, which self-assemble into three-dimensional objects with advantageous electronic properties, have been investigated. Systematic relations between the size, substitution and shape with function were found, which enabled the further optimization of the materials. Hand in hand with the development of suitable methods for visualization and processing, promising results were obtained for performance of nanoscale electronic devices. PMID- 17428767 TI - Hole-vibronic coupling in oligothiophenes: impact of backbone torsional flexibility on relaxation energies. AB - Density functional theory calculations together with highly resolved gas-phase ultraviolet photoelectron spectroscopy have been applied to oligothiophene chains with up to eight thiophene rings. One of the important parameters governing the charge transport properties in the condensed phase is the amount of energy relaxation upon ionization. Here, we investigate the impact on this parameter of the backbone flexibility present in oligothiophenes as a result of inter-ring torsional motions. With respect to oligoacenes that are characterized by a coplanar and rigid backbone, the torsional flexibility in oligothiophenes adds to the relaxation energy and leads to the broadening of the first ionization peak, making its analysis more complex. PMID- 17428769 TI - Supramolecular self-assembly codes for functional structures. AB - Small-molecule self-assembly has proven to be a rich field for the controlled synthesis of supramolecular objects with the size scale of polymers and interesting properties. Using several recent examples from our laboratory, we discuss the development of chemical structure codes for supramolecular self assembly objects with defined shapes. The resulting materials formed by these objects are promising for electronic functions and biological functions for regenerative medicine. PMID- 17428770 TI - An agent-based model of group decision making in baboons. AB - We present an agent-based model of the key activities of a troop of chacma baboons (Papio hamadryas ursinus) based on the data collected at De Hoop Nature Reserve in South Africa. We analyse the predictions of the model in terms of how well it is able to duplicate the observed activity patterns of the animals and the relationship between the parameters that control the agent's decision procedure and the model's predictions. At the current stage of model development, we are able to show that across a wide range of decision parameter values, the baboons are able to achieve their energetic and social time requirements. The simulation results also show that decisions concerning movement (group action selection) have the greatest influence on the outcomes. Those cases where the model's predictions fail to agree with the observed activity patterns have highlighted key elements that were missing from the field data, and that would need to be collected in subsequent fieldwork. Based on our experience, we believe group decision making is a fertile field for future research, and agent-based modelling offers considerable scope for understanding group action selection. PMID- 17428771 TI - Action selection and refinement in subcortical loops through basal ganglia and cerebellum. AB - Subcortical loops through the basal ganglia and the cerebellum form computationally powerful distributed processing modules (DPMs). This paper relates the computational features of a DPM's loop through the basal ganglia to experimental results for two kinds of natural action selection. First, functional imaging during a serial order recall task was used to study human brain activity during the selection of sequential actions from working memory. Second, microelectrode recordings from monkeys trained in a step-tracking task were used to study the natural selection of corrective submovements. Our DPM-based model assisted in the interpretation of puzzling data from both of these experiments. We come to posit that the many loops through the basal ganglia each regulate the embodiment of pattern formation in a given area of cerebral cortex. This operation serves to instantiate different kinds of action (or thought) mediated by different areas of cerebral cortex. We then use our findings to formulate a model of the aetiology of schizophrenia. PMID- 17428772 TI - Spatial models of political competition with endogenous political parties. AB - Two important human action selection processes are the choice by citizens of parties to support in elections and the choice by party leaders of policy 'packages' offered to citizens in order to attract this support. Having reviewed approaches analysing these choices and the reasons for doing this using the methodology of agent-based modelling, we extend a recent agent-based model of party competition to treat the number and identity of political parties as an output of, rather than an input to, the process of party competition. Party birth is modelled as an endogenous change of agent type from citizen to party leader, which requires describing citizen dissatisfaction with the history of the system. Endogenous birth and death of parties transforms into a dynamic system even in an environment where all agents have otherwise non-responsive adaptive rules. A key parameter is the survival threshold, with lower thresholds leaving citizens on average less dissatisfied. Paradoxically, the adaptive rule most successful for party leaders in winning votes makes citizens on average less happy than under other policy-selection rules. PMID- 17428773 TI - Biologically constrained action selection improves cognitive control in a model of the Stroop task. AB - The Stroop task is a paradigmatic psychological task for investigating stimulus conflict and the effect this has on response selection. The model of Cohen et al. (Cohen et al. 1990 Psychol. Rev. 97, 332-361) has hitherto provided the best account of performance in the Stroop task, but there remains certain key data that it fails to match. We show that this failure is due to the mechanism used to perform final response selection-one based on the diffusion model of choice behaviour (Ratcliff 1978 Psychol. Rev. 85, 59-108). We adapt the model to use a selection mechanism which is based on the putative human locus of final response selection, the basal ganglia/thalamo-cortical complex (Redgrave et al. 1999 Neuroscience 89, 1009-1023). This improves the match to the core human data and, additionally, makes it possible for the model to accommodate, in a principled way, additional mechanisms of cognitive control that enable better fits to the data. This work prompts a critique of the diffusion model as a mechanism of response selection, and the features that any response mechanism must possess to provide adaptive action selection. We conclude that the consideration of biologically constrained solutions to the action selection problem is vital to the understanding and improvement of cognitive models of response selection. PMID- 17428774 TI - Extending a biologically inspired model of choice: multi-alternatives, nonlinearity and value-based multidimensional choice. AB - The leaky competing accumulator (LCA) is a biologically inspired model of choice. It describes the processes of leaky accumulation and competition observed in neuronal populations during choice tasks and it accounts for reaction time distributions observed in psychophysical experiments. This paper discusses recent analyses and extensions of the LCA model. First, it reviews the dynamics and examines the conditions that make the model achieve optimal performance. Second, it shows that nonlinearities of the type present in biological neurons improve performance when the number of choice alternatives increases. Third, the model is extended to value-based choice, where it is shown that nonlinearities in the value function explain risk aversion in risky choice and preference reversals in choice between alternatives characterized across multiple dimensions. PMID- 17428775 TI - Understanding decision-making deficits in neurological conditions: insights from models of natural action selection. AB - Models of natural action selection implicate fronto-striatal circuits in both motor and cognitive 'actions'. Dysfunction of these circuits leads to decision making deficits in various populations. We review how computational models provide insights into the mechanistic basis for these deficits in Parkinson's patients and those with ventromedial frontal damage. We then consider implications of the models for understanding behaviour and cognition in attention deficit/hyperactivity disorder (ADHD). Incorporation of cortical noradrenaline function into the model improves action selection in noisy environments and accounts for response variability in ADHD. We close with more general clinical implications. PMID- 17428776 TI - Is there a brainstem substrate for action selection? AB - The search for the neural substrate of vertebrate action selection has focused on structures in the forebrain and midbrain, and particularly on the group of sub cortical nuclei known as the basal ganglia. Yet, the behavioural repertoire of decerebrate and neonatal animals suggests the existence of a relatively self contained neural substrate for action selection in the brainstem. We propose that the medial reticular formation (mRF) is the substrate's main component and review evidence showing that the mRF's inputs, outputs and intrinsic organization are consistent with the requirements of an action-selection system. The internal architecture of the mRF is composed of interconnected neuron clusters. We present an anatomical model which suggests that the mRF's intrinsic circuitry constitutes a small-world network and extend this result to show that it may have evolved to reduce axonal wiring. Potential configurations of action representation within the internal circuitry of the mRF are then assessed by computational modelling. We present new results demonstrating that each cluster's output is most likely to represent activation of a component action; thus, coactivation of a set of these clusters would lead to the coordinated behavioural response observed in the animal. Finally, we consider the potential integration of the basal ganglia and mRF substrates for selection and suggest that they may collectively form a layered/hierarchical control system. PMID- 17428777 TI - Multilevel structure in behaviour and in the brain: a model of Fuster's hierarchy. AB - A basic question, intimately tied to the problem of action selection, is that of how actions are assembled into organized sequences. Theories of routine sequential behaviour have long acknowledged that it must rely not only on environmental cues but also on some internal representation of temporal or task context. It is assumed, in most theories, that such internal representations must be organized into a strict hierarchy, mirroring the hierarchical structure of naturalistic sequential behaviour. This article reviews an alternative computational account, which asserts that the representations underlying naturalistic sequential behaviour need not, and arguably cannot, assume a strictly hierarchical form. One apparent liability of this theory is that it seems to contradict neuroscientific evidence indicating that different levels of sequential structure in behaviour are represented at different levels in a hierarchy of cortical areas. New simulations, reported here, show not only that the original computational account can be reconciled with this alignment between behavioural and neural organization, but also that it gives rise to a novel explanation for how this alignment might develop through learning. PMID- 17428778 TI - Towards an executive without a homunculus: computational models of the prefrontal cortex/basal ganglia system. AB - The prefrontal cortex (PFC) has long been thought to serve as an 'executive' that controls the selection of actions and cognitive functions more generally. However, the mechanistic basis of this executive function has not been clearly specified often amounting to a homunculus. This paper reviews recent attempts to deconstruct this homunculus by elucidating the precise computational and neural mechanisms underlying the executive functions of the PFC. The overall approach builds upon existing mechanistic models of the basal ganglia (BG) and frontal systems known to play a critical role in motor control and action selection, where the BG provide a 'Go' versus 'NoGo' modulation of frontal action representations. In our model, the BG modulate working memory representations in prefrontal areas to support more abstract executive functions. We have developed a computational model of this system that is capable of developing human-like performance on working memory and executive control tasks through trial-and-error learning. This learning is based on reinforcement learning mechanisms associated with the midbrain dopaminergic system and its activation via the BG and amygdala. Finally, we briefly describe various empirical tests of this framework. PMID- 17428780 TI - Compromise strategies for action selection. AB - Among the many properties suggested for action-selection mechanisms, a prominent one is the ability to select compromise actions, i.e. actions that are not the best to satisfy any active goal in isolation, but rather compromise between multiple goals. This paper briefly reviews the history of compromise behaviour and presents experimental analyses of it in an attempt to determine how much compromise behaviour aids an agent. It concludes that optimal compromise behaviour has a surprisingly small benefit over non-compromise behaviour in the experiments performed, and presents some reasons why this may be true and hypothesizes cases where compromise behaviour is truly useful. In particular, it hypothesizes that a crucial factor is the level at which an action is taken (low level actions are specific, such as 'move left leg'; high-level actions are vague, such as 'forage for food'). This paper hypothesizes that compromise behaviour is more beneficial for high- than low-level actions. PMID- 17428779 TI - Cortical mechanisms of action selection: the affordance competition hypothesis. AB - At every moment, the natural world presents animals with two fundamental pragmatic problems: selection between actions that are currently possible and specification of the parameters or metrics of those actions. It is commonly suggested that the brain addresses these by first constructing representations of the world on which to build knowledge and make a decision, and then by computing and executing an action plan. However, neurophysiological data argue against this serial viewpoint. In contrast, it is proposed here that the brain processes sensory information to specify, in parallel, several potential actions that are currently available. These potential actions compete against each other for further processing, while information is collected to bias this competition until a single response is selected. The hypothesis suggests that the dorsal visual system specifies actions which compete against each other within the fronto parietal cortex, while a variety of biasing influences are provided by prefrontal regions and the basal ganglia. A computational model is described, which illustrates how this competition may take place in the cerebral cortex. Simulations of the model capture qualitative features of neurophysiological data and reproduce various behavioural phenomena. PMID- 17428781 TI - The ecology of action selection: insights from artificial life. AB - The problem of action selection has two components: what is selected and how is it selected? To understand what is selected, it is necessary to distinguish between behavioural and mechanistic levels of description. Animals do not choose between behaviours per se; rather, behaviour reflects interactions among brains, bodies and environments. To understand what guides selection, it is useful to take a normative perspective that evaluates behaviour in terms of a fitness metric. This perspective, rooted in behavioural ecology, can be especially useful for understanding apparently irrational choice behaviour. This paper describes a series of models that use artificial life (AL) techniques to address the above issues. We show that successful action selection can arise from the joint activity of parallel, loosely coupled sensorimotor processes. We define a class of AL models that help to bridge the ecological approaches of normative modelling and agent- or individual-based modelling (IBM). Finally, we show how an instance of apparently suboptimal decision making, the matching law, can be accounted for by adaptation to competitive foraging environments. PMID- 17428782 TI - Do we expect natural selection to produce rational behaviour? AB - We expect that natural selection should result in behavioural rules which perform well; however, animals (including humans) sometimes make bad decisions. Researchers account for these with a variety of explanations; we concentrate on two of them. One explanation is that the outcome is a side effect; what matters is how a rule performs (in terms of reproductive success). Several rules may perform well in the environment in which they have evolved, but their performance may differ in a 'new' environment (e.g. the laboratory). Some rules may perform very badly in this environment. We use the debate about whether animals follow the matching law rather than maximizing their gains as an illustration. Another possibility is that we were wrong about what is optimal. Here, the general idea is that the setting in which optimal decisions are investigated is too simple and may not include elements that add extra degrees of freedom to the situation. PMID- 17428783 TI - Introduction. Modelling natural action selection. AB - Action selection is the task of resolving conflicts between competing behavioural alternatives. This theme issue is dedicated to advancing our understanding of the behavioural patterns and neural substrates supporting action selection in animals, including humans. The scope of problems investigated includes: (i) whether biological action selection is optimal (and, if so, what is optimized), (ii) the neural substrates for action selection in the vertebrate brain, (iii) the role of perceptual selection in decision-making, and (iv) the interaction of group and individual action selection. A second aim of this issue is to advance methodological practice with respect to modelling natural action section. A wide variety of computational modelling techniques are therefore employed ranging from formal mathematical approaches through to computational neuroscience, connectionism and agent-based modelling. The research described has broad implications for both natural and artificial sciences. One example, highlighted here, is its application to medical science where models of the neural substrates for action selection are contributing to the understanding of brain disorders such as Parkinson's disease, schizophrenia and attention deficit/hyperactivity disorder. PMID- 17428784 TI - Structure of the human lung cytochrome P450 2A13. AB - The human lung cytochrome P450 2A13 (CYP2A13) activates the nicotine-derived procarcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) into DNA altering compounds that cause lung cancer. Another cytochrome P450, CYP2A6, is also present in human lung, but at much lower levels. Although these two enzymes are 93.5% identical, CYP2A13 metabolizes NNK with much lower K(m) values than does CYP2A6. To investigate the structural differences between these two enzymes the structure of CYP2A13 was determined to 2.35A by x-ray crystallography and compared with structures of CYP2A6. As expected, the overall CYP2A13 and CYP2A6 structures are very similar with an average root mean square deviation of 0.5A for the Calpha atoms. Like CYP2A6, the CYP2A13 active site cavity is small and highly hydrophobic with a cluster of Phe residues composing the active site roof. Active site residue Asn(297) is positioned to hydrogen bond with an adventitious ligand, identified as indole. Amino acid differences between CYP2A6 and CYP2A13 at positions 117, 300, 301, and 208 relate to different orientations of the ligand plane in the two protein structures and may underlie the significant variations observed in binding and catalysis of many CYP2A ligands. In addition, docking studies suggest that residues 365 and 366 may also contribute to differences in NNK metabolism. PMID- 17428785 TI - Defining paxilline biosynthesis in Penicillium paxilli: functional characterization of two cytochrome P450 monooxygenases. AB - Indole diterpenes are a large, structurally and functionally diverse group of secondary metabolites produced by filamentous fungi. Biosynthetic schemes have been proposed for these metabolites but until recently none of the proposed steps had been validated by biochemical or genetic studies. Using Penicillium paxilli as a model experimental system to study indole diterpene biosynthesis we previously showed by deletion analysis that a cluster of seven genes is required for paxilline biosynthesis. Two of these pax genes, paxP and paxQ (encoding cytochrome P450 monooxygenases), are required in the later steps in this pathway. Here, we describe the function of paxP and paxQ gene products by feeding proposed paxilline intermediates to strains lacking the pax cluster but containing ectopically integrated copies of paxP or paxQ. Transformants containing paxP converted paspaline into 13-desoxypaxilline as the major product and beta-PC-M6 as the minor product. beta-PC-M6, but not alpha-PC-M6, was also a substrate for PaxP and was converted to 13-desoxypaxilline. paxQ-containing transformants converted 13-desoxypaxilline into paxilline. These results confirm that paspaline, beta-PC-M6, and 13-desoxypaxilline are paxilline intermediates and that paspaline and beta-PC-M6 are substrates for PaxP, and 13-desoxypaxilline is a substrate for PaxQ. PaxP and PaxQ also utilized beta-paxitriol and alpha-PC-M6 as substrates converting them to paxilline and alpha-paxitriol, respectively. These findings have allowed us to delineate clearly the biosynthetic pathway for paxilline for the first time. PMID- 17428786 TI - The structure of 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase provides insights into the mechanism of uric acid degradation. AB - The complete degradation of uric acid to (S)-allantoin, as recently elucidated, involves three enzymatic reactions. Inactivation by pseudogenization of the genes of the pathway occurred during hominoid evolution, resulting in a high concentration of urate in the blood and susceptibility to gout. Here, we describe the 1.8A resolution crystal structure of the homodimeric 2-oxo-4-hydroxy-4 carboxy-5-ureidoimidazoline decarboxylase, which catalyzes the last step in the urate degradation pathway, for both ligand-free enzyme and enzyme in complex with the substrate analogs (R)-allantoin and guanine. Each monomer comprises ten alpha helices, grouped into two domains and assembled in a novel fold. The structure and the mutational analysis of the active site have allowed us to identify some residues that are essential for catalysis, among which His-67 and Glu-87 appear to play a particularly significant role. Glu-87 may facilitate the exit of the carboxylate group because of electrostatic repulsion that destabilizes the ground state of the substrate, whereas His-67 is likely to be involved in a protonation step leading to the stereoselective formation of the (S)-allantoin enantiomer as reaction product. The structural and functional characterization of 2-oxo-4 hydroxy-4-carboxy-5-ureidoimidazoline decarboxylase can provide useful information in view of the potential use of this enzyme in the enzymatic therapy of gout. PMID- 17428787 TI - Characterization of a novel cell wall-anchored protein with carboxylesterase activity required for virulence in Mycobacterium tuberculosis. AB - Pooled mutant competition assays have shown that the Mycobacterium tuberculosis MT2282 gene (Rv2224c, annotated as encoding a proteinase) is required for bacterial survival in mice. To understand the mechanism of this requirement, we conducted a genetic and biochemical study of the MT2282 gene and its product. MT2282 encodes a member of the microbial esterase/lipase family with active site consensus sequences of G-X-S-X-G, and we have concluded that the MT2282 protein is, in fact, a cell wall-associated carboxylesterase rather than a proteinase, as initially annotated. The MT2282 gene product preferentially hydrolyzes ester bonds of substrates with intermediate carbon chain length. Purified MT2282 is a monomer with enzymatic catalysis properties that fit in the Michaelis-Menten kinetic model. Esterase activity was inhibited by paraoxon and dichlorvos. Replacement of Ser215, Asp450, and His477 by Ala in the consensus motifs completely abolishes esterase activity, suggesting that Ser215-Asp450-His477 forms a catalytic triad with Ser215 as an active site residue. To evaluate the role of the MT2282 in pathogenesis, the gene was deleted from the M. tuberculosis genome. BALB/c mouse aerosol infections showed reduced colony-forming unit loads in lungs and spleens and less lung pathology for the DeltaMT2282 mutant. High dose intravenous infection of mice with the mutant resulted in a significantly delayed time to death compared with the wild type or complemented mutant. These results indicate that MT2282 encodes a cell wall-associated carboxylesterase, which is required for full virulence of M. tuberculosis. We propose that MT2282 (Rv2224c) and its adjacent paralogous gene MT2281 (Rv2223c) be named caeA and caeB respectively, for carboxylesterase A and B. PMID- 17428788 TI - Overlapping roles of the methylated DNA-binding protein MBD1 and polycomb group proteins in transcriptional repression of HOXA genes and heterochromatin foci formation. AB - Methylated DNA binding domain (MBD) proteins and Polycomb group (PcG) proteins maintain epigenetic silencing of transcriptional activity. We report that the DNA methylation-mediated repressor MBD1 interacts with Ring1b and hPc2, the major components of Polycomb repressive complex 1. The cysteine-rich CXXC domains of MBD1 bound to Ring1b and the chromodomain of hPc2. Chromatin immunoprecipitation analysis revealed that MBD1 and hPc2 were present in silenced Homeobox A (HOXA) genes which could be reactivated by knockdown of either MBD1 or hPc2, suggesting that MBD1 and hPc2 cooperate for transcriptional repression of HOXA genes. In the nuclei of HeLa cells, MBD1 existed in close association with these PcG proteins in some heterochromatin foci, whereas an MBD1 mutant lacking the CXXC domains or an hPc2 mutant lacking the chromodomain lost this colocalization in foci. Use of the DNA demethylating agent 5-azadeoxycytidine abolished the formation of MBD1 foci but not PcG foci. Knockdown of MBD1 by small interfering RNAs did not affect the foci containing hPc2 and Ring1b, whereas the MBD1 foci were not influenced by knockdown of hPc2. These indicate that the heterochromatin foci showing MBD1 and hPc2 colocalization arise through the interaction of MBD1 and hPc2 and that the foci of MBD1 are separable from those of the PcG proteins per se. Our present findings suggest that MBD1 and PcG proteins have overlapping roles in epigenetic gene silencing and heterochromatin foci formation through their interactions. PMID- 17428789 TI - Phosphatidylethanolamine, a limiting factor of autophagy in yeast strains bearing a defect in the carboxypeptidase Y pathway of vacuolar targeting. AB - Vps4p and Vps36p of Saccharomyces cerevisiae are involved in the transport of proteins to the vacuole via the carboxypeptidase Y pathway. We found that deletion of VPS4 and VPS36 caused impaired maturation of the vacuolar proaminopeptidase I (pAPI) via autophagy or the cytosol to vacuole targeting pathway. Supplementation with ethanolamine rescued this defect, leading to an increase of the cellular amount of phosphatidylethanolamine (PtdEtn), an enhanced level of the PtdEtn-binding autophagy protein Atg8p and a balanced rate of autophagy. We also discovered that maturation of pAPI was generally affected by PtdEtn depletion in a psd1Delta psd2Delta mutant due to reduced recruitment of Atg8p to the preautophagosomal structure. Ethanolamine supplementation provided the necessary amounts of PtdEtn for complete maturation of pAPI. Since the expression level of Atg8p was not compromised in the psd1Delta psd2Delta strain, we concluded that the amount of available PtdEtn was limiting. Thus, PtdEtn appears to be a limiting factor for the balance of the carboxypeptidase Y pathway and autophagy/the cytosol to vacuole targeting pathway in the yeast. PMID- 17428790 TI - Potential role for phosphorylation in differential regulation of the assembly of dynein light chains. AB - The homodimeric light chains LC8 and Tctex-1 are integral parts of the microtubule motor cytoplasmic dynein, as they directly associate with dynein intermediate chain IC and various cellular cargoes. These light chains appear to regulate assembly of the dynein complex by binding to and promoting dimerization of IC. In addition, both LC8 and Tctex-1 play roles in signaling, apoptosis, and neuronal development that are independent of their function in dynein, but it is unclear how these various activities are modulated. Both light chains undergo specific phosphorylation, and here we present biochemical and NMR analyses of phosphomimetic mutants that indicate how phosphorylation may regulate light chain function. For both LC8 and Tctex-1, phosphorylation promotes dissociation from IC while retaining their binding activity with other non-dynein proteins. Although LC8 and Tctex-1 are homologs having a common fold, their reduced affinity for IC upon phosphorylation arises by different mechanisms. In the case of Tctex-1, phosphorylation directly masks the IC binding site at the dimer interface, whereas for LC8, phosphorylation dissociates the dimer and indirectly eliminates the binding site. This modulation of the monomer-dimer equilibrium by phosphorylation provides a novel mechanism for discrimination among LC8 binding partners. PMID- 17428791 TI - Crystal structures and site-directed mutagenesis of a mycothiol-dependent enzyme reveal a novel folding and molecular basis for mycothiol-mediated maleylpyruvate isomerization. AB - Mycothiol (MSH) is the major low molecular mass thiols in many Gram-positive bacteria such as Mycobacterium tuberculosis and Corynebacterium glutamicum. The physiological roles of MSH are believed to be equivalent to those of GSH in Gram negative bacteria, but current knowledge of MSH is limited to detoxification of alkalating chemicals and protection from host cell defense/killing systems. Recently, an MSH-dependent maleylpyruvate isomerase (MDMPI) was discovered from C. glutamicum, and this isomerase represents one example of many putative MSH dependent enzymes that take MSH as cofactor. In this report, fourteen mutants of MDMPI were generated. The wild type and mutant (H52A) MDMPIs were crystallized and their structures were solved at 1.75 and 2.05 A resolution, respectively. The crystal structures reveal that this enzyme contains a divalent metal-binding domain and a C-terminal domain possessing a novel folding pattern (alphabetaalphabetabetaalpha fold). The divalent metal-binding site is composed of residues His52, Glu144, and His148 and is located at the bottom of a surface pocket. Combining the structural and site-directed mutagenesis studies, it is proposed that this surface pocket including the metal ion and MSH moiety formed the putative catalytic center. PMID- 17428792 TI - Ataxia telangiectasia mutated (ATM) signaling network is modulated by a novel poly(ADP-ribose)-dependent pathway in the early response to DNA-damaging agents. AB - Poly(ADP-ribosyl)ation is a post-translational modification that is instantly stimulated by DNA strand breaks creating a unique signal for the modulation of protein functions in DNA repair and cell cycle checkpoint pathways. Here we report that lack of poly(ADP-ribose) synthesis leads to a compromised response to DNA damage. Deficiency in poly(ADP-ribosyl)ation metabolism induces profound cellular sensitivity to DNA-damaging agents, particularly in cells deficient for the protein kinase ataxia telangiectasia mutated (ATM). At the biochemical level, we examined the significance of poly(ADP-ribose) synthesis on the regulation of early DNA damage-induced signaling cascade initiated by ATM. Using potent PARP inhibitors and PARP-1 knock-out cells, we demonstrate a functional interplay between ATM and poly(ADP-ribose) that is important for the phosphorylation of p53, SMC1, and H2AX. For the first time, we demonstrate a functional and physical interaction between the major DSB signaling kinase, ATM and poly(ADP ribosyl)ation by PARP-1, a key enzyme of chromatin remodeling. This study suggests that poly(ADP-ribose) might serve as a DNA damage sensory molecule that is critical for early DNA damage signaling. PMID- 17428793 TI - Structural basis of Na+ activation mimicry in murine thrombin. AB - Unlike human thrombin, murine thrombin lacks Na+ activation due to the charge reversal substitution D222K in the Na+ binding loop. However, the enzyme is functionally stabilized in a Na+-bound form and is highly active toward physiologic substrates. The structural basis of this peculiar property is unknown. Here, we present the 2.2 A resolution x-ray crystal structure of murine thrombin in the absence of inhibitors and salts. The enzyme assumes an active conformation, with Ser-195, Glu-192, and Asp-189 oriented as in the Na+-bound fast form of human thrombin. Lys-222 completely occludes the pore of entry to the Na+ binding site and positions its side chain inside the pore, with the Nzeta atom H-bonded to the backbone oxygen atoms of Lys-185, Asp-186b, and Lys-186d. The same architecture is observed in the 1.75 A resolution structure of a thrombin chimera in which the human enzyme carries all residues defining the Na+ pore in the murine enzyme. These findings demonstrate that Na+ activation in thrombin is linked to the architecture of the Na+ pore. The molecular strategy of Na+ activation mimicry unraveled for murine thrombin is relevant to serine proteases and enzymes activated by monovalent cations in general. PMID- 17428794 TI - Regulation of type II collagen expression by histone deacetylase in articular chondrocytes. AB - Histone deacetylase (HDAC) regulates various cellular processes by modulating gene expression. Here, we investigated the role of HDAC in the expression of type II collagen, a marker of differentiated chondrocytes. We found that HDAC activity in primary articular chondrocytes decreases during dedifferentiation induced by serial monolayer culture and that the activity recovered during redifferentiation induced by three-dimensional culture in a cell pellet. Inhibition of HDAC with trichostatin A or PXD101 was sufficient to block type II collagen expression in primary culture chondrocytes. HDAC inhibition also blocked the redifferentiation of dedifferentiated chondrocytes by suppressing the synthesis and accumulation of type II collagen. HDAC inhibition promoted the expression of Wnt-5a, which is known to inhibit type II collagen expression, and knockdown of Wnt-5a blocked the ability of HDAC inhibitors to suppress type II collagen expression. In addition, the induction of Wnt-5a expression by HDAC inhibitors was associated with acetylation of the Wnt-5a promoter. Taken together, our results suggest that HDAC promotes type II collagen expression by suppressing the transcription of Wnt-5a. PMID- 17428795 TI - Ligand binding and calcium influx induce distinct ectodomain/gamma-secretase processing pathways of EphB2 receptor. AB - Binding of EphB receptors to ephrinB ligands on the surface of adjacent cells initiates signaling cascades that regulate angiogenesis, axonal guidance, and neuronal plasticity. These functions require processing of EphB receptors and removal of EphB-ephrinB complexes from the cell surface, but the mechanisms involved are poorly understood. Here we show that the ectodomain of EphB2 receptor is released to extracellular space following cleavage after EphB2 residue 543. The remaining membrane-associated fragment is cleaved by the presenilin-dependent gamma-secretase activity after EphB2 residue 569 releasing an intracellular peptide that contains the cytoplasmic domain of EphB2. This cleavage is inhibited by presenilin 1 familial Alzheimer disease mutations. Processing of EphB2 receptor depends on specific treatments: ephrinB ligand induced processing requires endocytosis, and the ectodomain cleavage is sensitive to peptide inhibitor N-benzyloxycarbonyl-Val-Leu-leucinal but insensitive to metalloproteinase inhibitor GM6001. The ligand-induced processing takes place in endosomes and involves the rapid degradation of the extracellular EphB2. EphrinB ligand stimulates ubiquitination of EphB2 receptor. Calcium influx- and N-methyl d-aspartic acid-induced processing of EphB2 is inhibited by GM6001 and ADAM10 inhibitors but not by N-benzyloxycarbonyl-Val-Leu-leucinal. This processing requires no endocytosis and promotes rapid shedding of extracellular EphB2, indicating that it takes place at the plasma membrane. Our data identify novel cleavages and modifications of EphB2 receptor and indicate that specific conditions determine the proteolytic systems and subcellular sites involved in the processing of this receptor. PMID- 17428796 TI - Tissue-specific mechanisms for CCN2/CTGF persistence in fibrotic gingiva: interactions between cAMP and MAPK signaling pathways, and prostaglandin E2-EP3 receptor mediated activation of the c-JUN N-terminal kinase. AB - Prostaglandin E(2) blocks transforming growth factor TGF beta1-induced CCN2/CTGF expression in lung and kidney fibroblasts. PGE(2) levels are high in gingival tissues yet CCN2/CTGF expression is elevated in fibrotic gingival overgrowth. Gingival fibroblast expression of CCN2/CTGF in the presence of PGE(2) led us to compare the regulation of CCN2/CTGF expression in fibroblasts cultured from different tissues. Data demonstrate that the TGFbeta1-induced expression of CCN2/CTGF in human lung and renal mesangial cells is inhibited by 10 nm PGE(2), whereas human gingival fibroblasts are resistant. Ten nm PGE(2) increases cAMP accumulation in lung but not gingival fibroblasts, which require 1 mum PGE(2) to elevate cAMP. Micromolar PGE(2) only slightly reduces the TGFbeta1-stimulated CCN2/CTGF levels in gingival cells. EP2 prostaglandin receptor activation with butaprost blocks the TGFbeta1-stimulated expression of CCN2/CTGF expression in lung, but not gingival, fibroblasts. In lung fibroblasts, inhibition of the TGFbeta1-stimulated CCN2/CTGF by PGE(2), butaprost, or forskolin is due to p38, ERK, and JNK MAP kinase inhibition that is cAMP-dependent. Inhibition of any two MAPKs completely blocks CCN2/CTGF expression stimulated by TGFbeta1. These data mimic the inhibitory effects of 10 nm PGE(2) and forskolin that were dependent on PKA activity. In gingival fibroblasts, the sole MAPK mediating the TGFbeta1 stimulated CCN2/CTGF expression is JNK. Whereas forskolin reduces TGFbeta1 stimulated expression of CCN2/CTGF by 35% and JNK activation in gingival fibroblasts, micromolar PGE(2)-stimulated JNK in gingival fibroblasts and opposes the inhibitory effects of cAMP on CCN2/CTGF expression. Stimulation of the EP3 receptor with sulprostone results in a robust increase in JNK activation in these cells. Taken together, data identify two mechanisms by which TGFbeta1-stimulated CCN2/CTGF levels in human gingival fibroblasts resist down-regulation by PGE(2): (i) cAMP cross-talk with MAPK pathways is limited in gingival fibroblasts; (ii) PGE(2) activation of the EP3 prostanoid receptor stimulates the activation of JNK. PMID- 17428797 TI - The phosphorylation state of GluR1 subunits determines the susceptibility of AMPA receptors to calpain cleavage. AB - The alpha-Amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid receptor (AMPAR) is an ionotropic glutamate receptor that governs most of excitatory synaptic transmission in neurons. In vitro biochemical assay has shown that calpain, a Ca2+-activated protease, can cleave AMPAR GluR1 subunits. Our physiological study found that calpain, which was activated by prolonged stimulation of the N-methyl D-aspartate receptor (100 microM, 10 min), caused a substantial suppression of AMPAR currents in cortical neurons. Since the phosphorylation sites of GluR1 by several protein kinases are located in close proximity to the calpain cleavage sites, we investigated the effect of phosphorylation on the susceptibility of GluR1 to calpain cleavage. Interestingly, we found that the calpain regulation of AMPAR currents was diminished by inhibition of Ca2+/calmodulin-dependent protein kinase II (CaMKII) but was augmented by inhibition of protein phosphatase 1/2A (PP1/2A). In agreement with this, in vitro assay showed that the calpain-induced proteolytic cleavage of GluR1 C-terminal fusion protein was strongly potentiated by adding the purified active CaMKII, and GluR1 phosphorylated at Ser831 by CaMKII is much more sensitive to calpain cleavage. Taken together, our data suggest that calpain activation suppresses AMPA receptor currents via proteolytic cleavage of GluR1 subunits, and the susceptibility of AMPARs to calpain cleavage is determined by the phosphorylation state of GluR1 subunits, which is mediated by CaMKII-PP1/2A activity. PMID- 17428798 TI - Solution structure of human secretory component and implications for biological function. AB - Secretory component (SC) in association with polymeric IgA (pIgA) forms secretory IgA, the major antibody active at mucosal surfaces. SC also exists in the free form, with innate-like neutralizing properties against pathogens. Free SC consists of five glycosylated variable (V)-type Ig domains (D1-D5), whose structure was determined by x-ray and neutron scattering, ultracentrifugation, and modeling. With a radius of gyration of 3.53-3.63 nm, a length of 12.5 nm, and a sedimentation coefficient of 4.0 S, SC possesses an unexpected compact structure. Constrained scattering modeling based on up to 13,000 trial models shows that SC adopts a J-shaped structure in which D4 and D5 are folded back against D2 and D3. The seven glycosylation sites are located on one side of SC, leaving known IgA-binding motifs free to interact with pIgA. This work represents the first analysis of the three-dimensional structure of full-length free SC and paves the way to a better understanding of the association between SC and its potential ligands, i.e. pIgA and pathogenic-associated motifs. PMID- 17428799 TI - Beta-globin active chromatin Hub formation in differentiating erythroid cells and in p45 NF-E2 knock-out mice. AB - Expression of the beta-globin genes proceeds from basal to exceptionally high levels during erythroid differentiation in vivo. High expression is dependent on the locus control region (LCR) and coincides with more frequent LCR-gene contacts. These contacts are established in the context of an active chromatin hub (ACH), a spatial chromatin configuration in which the LCR, together with other regulatory sequences, loops toward the active beta-globin-like genes. Here, we used recently established I/11 cells as a model system that faithfully recapitulates the in vivo erythroid differentiation program to study the molecular events that accompany and underlie ACH formation. Upon I/11 cell induction, histone modifications changed, the ACH was formed, and the beta-globin like genes were transcribed at rates similar to those observed in vivo. The establishment of frequent LCR-gene contacts coincided with a more efficient loading of polymerase onto the beta-globin promoter. Binding of the transcription factors GATA-1 and EKLF to the locus, although previously shown to be required, was not sufficient for ACH formation. Moreover, we used knock-out mice to show that the erythroid transcription factor p45 NF-E2, which has been implicated in beta-globin gene regulation, is dispensable for beta-globin ACH formation. PMID- 17428800 TI - Tau aggregation and toxicity in a cell culture model of tauopathy. AB - Intracellular aggregation of the microtubule-associated protein tau into filamentous inclusions is a defining characteristic of Alzheimer disease. Because appearance of tau-aggregate bearing lesions correlates with both cognitive decline and neurodegeneration, it has been hypothesized that tau aggregation may be directly toxic to cells that harbor them. Testing this hypothesis in cell culture has been complicated by the resistance of full-length tau isoforms to aggregation over experimentally tractable time periods. To overcome this limitation, a small-molecule agonist of the tau aggregation reaction, Congo red, was used to drive aggregation within HEK-293 cells expressing full-length tau isoform htau40. Formation of detergent-insoluble aggregates was both time and agonist concentration dependent. At 10 microM Congo red, detergent-insoluble aggregates appeared with pseudo-first order kinetics and a half-life of approximately 5 days. By 7 days in culture, total tau levels increased 2-fold, with approximately 30% of total tau converted into detergent-insoluble aggregates. Agonist addition also led to rapid losses in the tubulin binding activity of tau, although tau was not hyperphosphorylated as judged by occupancy of phosphorylation sites Ser396/Ser404. Tau aggregation was associated with decreased viability as detected by ToPro-3 uptake. The results, which establish a new approach for analysis of tau aggregation in cells independent of tau hyperphosphorylation, suggest that conformational changes associated with aggregation are incompatible with microtubule binding, and that toxicity associated with intracellular tau aggregation is not acute but develops over a period of days. PMID- 17428801 TI - CARMA1 coiled-coil domain is involved in the oligomerization and subcellular localization of CARMA1 and is required for T cell receptor-induced NF-kappaB activation. AB - T lymphocyte (T cell) activation and proliferation is induced by the activation of multiple signal transduction pathways. Earlier studies indicate that CARMA1, a Caspase Recruitment Domain (CARD) and Membrane-associated GUanylate Kinase domain (MAGUK)-containing scaffold protein, plays an essential role in NF-kappaB activation induced by the costimulation of T cell receptor (TCR) and CD28 molecules. However, the molecular mechanism by which CARMA1 mediates TCR-CD28 costimulation-induced NF-kappaB activation is not fully understood. Here we show that CARMA1 is constitutively oligomerized. This oligomerization of CARMA1 is through its Coiled-coil domain. Disruption of the predicted structure of the Coiled-coil domain of CARMA1 impaired its oligomerization and, importantly, abrogated CARMA1-mediated NF-kappaB activation. Interestingly, disruption of the CC1 domain abrogates CARMA1 localization, whereas disruption of the CC2 domain seems to inhibit CARMA1 self-association. Together, our results demonstrate that the oligomerization of CARMA1 is required for TCR-induced NF-kappaB activation. PMID- 17428802 TI - RNA binding-independent dimerization of adenosine deaminases acting on RNA and dominant negative effects of nonfunctional subunits on dimer functions. AB - RNA editing that converts adenosine to inosine in double-stranded RNA (dsRNA) is mediated by adenosine deaminases acting on RNA (ADAR). ADAR1 and ADAR2 form respective homodimers, and this association is essential for their enzymatic activities. In this investigation, we set out experiments aiming to determine whether formation of the homodimer complex is mediated by an amino acid interface made through protein-protein interactions of two monomers or via binding of the two subunits to a dsRNA substrate. Point mutations were created in the dsRNA binding domains (dsRBDs) that abolished all RNA binding, as tested for two classes of ADAR ligands, long and short dsRNA. The mutant ADAR dimer complexes were intact, as demonstrated by their ability to co-purify in a sequential affinity-tagged purification and also by their elution at the dimeric fraction position on a size fractionation column. Our results demonstrated ADAR dimerization independent of their binding to dsRNA, establishing the importance of protein-protein interactions for dimer formation. As expected, these mutant ADARs could no longer perform their catalytic function due to the loss in substrate binding. Surprisingly, a chimeric dimer consisting of one RNA binding mutant monomer and a wild type partner still abolished its ability to bind and edit its substrate, indicating that ADAR dimers require two subunits with functional dsRBDs for binding to a dsRNA substrate and then for editing activity to occur. PMID- 17428803 TI - Mammalian Sec16/p250 plays a role in membrane traffic from the endoplasmic reticulum. AB - Coat protein complex II (COPII)-coated vesicles/carriers, which mediate export of proteins from the endoplasmic reticulum (ER), are formed at special ER subdomains in mammals, termed ER exit sites or transitional ER. The COPII coat consists of a small GTPase, Sar1, and two protein complexes, Sec23-Sec24 and Sec13-Sec31. Sec23 Sec24 and Sec13-Sec31 appear to constitute the inner and the outermost layers of the COPII coat, respectively. We previously isolated two mammalian proteins (p125 and p250) that bind to Sec23. p125 was found to be a mammalian-specific, phospholipase A(1)-like protein that participates in the organization of ER exit sites. Here we show that p250 is encoded by the KIAA0310 clone and has sequence similarity to yeast Sec16 protein. Although KIAA0310p was found to be localized at ER exit sites, subcellular fractionation revealed its predominant presence in the cytosol. Cytosolic KIAA0310p was recruited to ER membranes in a manner dependent on Sar1. Depletion of KIAA0310p mildly caused disorganization of ER exit sites and delayed protein transport from the ER, suggesting its implication in membrane traffic out of the ER. Overexpression of KIAA0310p affected ER exit sites in a manner different from that of p125. Binding experiments suggested that KIAA0310p interacts with both the inner and the outermost layer coat complexes, whereas p125 binds principally to the inner layer complex. Our results suggest that KIAA0310p, a mammalian homologue of yeast Sec16, builds up ER exit sites in cooperation with p125 and plays a role in membrane traffic from the ER. PMID- 17428804 TI - chi-Conotoxin and tricyclic antidepressant interactions at the norepinephrine transporter define a new transporter model. AB - Monoamine neurotransmitter transporters for norepinephrine (NE), dopamine and serotonin are important targets for antidepressants and analgesics. The conopeptide chi-MrIA is a noncompetitive and highly selective inhibitor of the NE transporter (NET) and is being developed as a novel intrathecal analgesic. We used site-directed mutagenesis to generate a suite of mutated transporters to identify two amino acids (Leu(469) and Glu(382)) that affected the affinity of chi-MrIA to inhibit [(3)H]NE uptake through human NET. Residues that increased the K(d) of a tricyclic antidepressant (nisoxetine) were also identified (Phe(207), Ser(225), His(296), Thr(381), and Asp(473)). Phe(207), Ser(225), His(296), and Thr(381) also affected the rate of NE transport without affecting NE K(m). In a new model of NET constructed from the bLeuT crystal structure, chi MrIA-interacting residues were located at the mouth of the transporter near residues affecting the binding of small molecule inhibitors. PMID- 17428805 TI - Noncovalent binding of small ubiquitin-related modifier (SUMO) protease to SUMO is necessary for enzymatic activities and cell growth. AB - SUMO proteases possess two enzymatic activities to hydrolyze the C-terminal region of SUMOs (hydrolase activity) and to remove SUMO from SUMO-conjugated substrates (isopeptidase activity). SUMO proteases bind to SUMOs noncovalently, but the physiological roles of the binding in the functions of SUMO proteases are not well understood. In this study we found that SUMO proteases (Axam, SENP1, and yeast Ulp1) show different preferences for noncovalent binding to various SUMOs (SUMO-1, -2, -3, and yeast Smt3) and that the hydrolase and isopeptidase activities of SUMO proteases are dependent on their binding to SUMOs through salt bridge. Expression of Smt3 suppressed the phenotype of yeast mutant lacking smt3, which exhibits growth arrest, and the binding of Ulp1 to Smt3 was essential for this rescue activity. Although expression of an Smt3 mutant (smt3R64E(GG)), which conjugates to substrate but loses the ability to bind to Ulp1, rescued the phenotype of yeast lacking smt3 partially, the mutant cells showed an increment in the doubling time and a delay of desumoylation of Smt3-conjugated Cdc3. These results indicate that the noncovalent binding of SUMO protease to SUMO through salt bridge is essential for the enzymatic activities and that the balance between sumoylation and desumoylation is important for cell growth control. PMID- 17428806 TI - Transcriptional silencing of nonsense codon-containing immunoglobulin micro genes requires translation of its mRNA. AB - Eukaryotes have evolved quality control mechanisms that prevent the expression of genes in which the protein coding potential is crippled by the presence of a premature translation-termination codon (PTC). In addition to nonsense-mediated mRNA decay (NMD), a well documented posttranscriptional consequence of the presence of a PTC in an mRNA, we recently reported the transcriptional silencing of PTC-containing immunoglobulin (Ig) mu and gamma minigenes when they are stably integrated into the genome of HeLa cells. Here we demonstrate that this transcriptional silencing of PTC-containing Ig-mu constructs requires active translation of the cognate mRNA, as it is not observed under conditions where translation of the PTC-containing mRNA is inhibited through an iron-responsive element in the 5'-untranslated region. Furthermore, RNA interference-mediated depletion of the essential NMD factor Upf1 not only abolishes NMD but also reduces the extent of nonsense-mediated transcriptional gene silencing (NMTGS). Collectively, our data indicate that NMTGS and NMD are linked, relying on the same mechanism for PTC recognition, and that the NMTGS pathway branches from the NMD pathway at a step after Upf1 function. PMID- 17428807 TI - dp5/HRK is a c-Jun target gene and required for apoptosis induced by potassium deprivation in cerebellar granule neurons. AB - In cerebellar granule neurons, a BH3-only Bcl-2 family member, death protein 5/harakiri, is up-regulated in a JNK-dependent manner during apoptosis induced by potassium deprivation. However, it is not clear whether c-Jun is directly involved in the induction of dp5. Here, we showed that the up-regulation of dp5, but not fas ligand and bim, after potassium deprivation was suppressed by the expression of a dominant negative form of c-Jun. Deletion analysis of the 5' flanking sequence of the dp5 gene revealed that a major responsive element responsible for the induction by potassium deprivation is an ATF binding site located at -116 to -109 relative to the transcriptional start site. Mutation of this site completely abolished promoter activation. Furthermore, a gel shift assay showed that a specific complex containing c-Jun and ATF2 recognized this site and increased in potassium-deprived cerebellar granule neurons. Chromatin immunoprecipitation demonstrated that c-Jun was able to bind to this site in vivo. Finally, we demonstrated that knockdown of Dp5 by small interfering RNA rescued neurons from potassium deprivation-induced apoptosis. Taken together, these results suggest that dp5 is a target gene of c-Jun and plays a critical role in potassium deprivation-induced apoptosis in cerebellar granule neurons. PMID- 17428808 TI - Anti-desmoglein 3 (Dsg3) monoclonal antibodies deplete desmosomes of Dsg3 and differ in their Dsg3-depleting activities related to pathogenicity. AB - Pemphigus vulgaris (PV) is an autoimmune blistering disease, characterized by the loss of cell-cell adhesion between epidermal keratinocytes and the presence of autoantibody against desmoglein 3 (Dsg3), which provides adhesive integrity to desmosomes between adjacent keratinocytes. We have previously shown that PV-IgG purified from patients depletes desmosomes of Dsg3. However, PV-IgG contains not only antibodies against a variety of different epitopes of Dsg3 but also against other unknown antigens. Therefore, we examined whether the Dsg3-depleting activity of PV-IgG is generated specifically by anti-Dsg3 activity in a human squamous cell carcinoma cell line (DJM-1) and normal human keratinocytes by using four different pathogenic and nonpathogenic monoclonal antibodies against Dsg3. We demonstrate that these monoclonal antibodies deplete cells and desmosomes of Dsg3, as PV-IgG does. Individual monoclonal anti-Dsg3 antibodies display characteristic limits to their Dsg3-depleting activity, which correlates with their pathogenic activities. In combination, these antibodies exert a cumulative or synergistic effect, which may explain the potent Dsg3-depleting capability of PV-IgG, which is polyclonal. Finally, although Dsg3-depletion activity correlated with AK-monoclonal antibody pathogenicity in mouse models, the residual level of Dsg3, when below approximately 50%, does not correlate with the adhesive strength index in the present study. This may suggest that although the Dsg3 depletion is not indicative for adhesive strength, the level of Dsg3 can be used as a read-out of pathogenic changes within the cell and that the Dsg3 depletion from desmosomes plays an important role in skin fragility or susceptibility to blister formation in PV patients. PMID- 17428809 TI - Commentary: back to basics--good news from a birthplace of DOTS. PMID- 17428810 TI - Effect of recombinant human activated protein C on the bactericidal activity of human monocytes and modulation of pro-inflammatory cytokines in the presence of antimicrobial agents. AB - OBJECTIVES: To determine the effects of recombinant human activated protein C (rhAPC) on the antimicrobial activity and cytokine production of normal human monocyte-derived macrophages (MDMs) in the presence and absence of Escherichia coli infection, with and without treatment with levofloxacin or ampicillin. METHODS: MDM monolayers were infected with E. coli ATCC 25922 and treated with levofloxacin or ampicillin in the presence or absence of rhAPC. Antimicrobial activity and cytokine (TNF-alpha, IL-1beta, IL-6 and IL-8) concentrations in the supernatants were measured. RESULTS: When low concentrations of levofloxacin were used, a therapeutic concentration of rhAPC enhanced intracellular antibacterial activity at all time points. With ampicillin, antibacterial activity increased, was unaffected or diminished depending upon the drug concentration and assay time. Without antibiotics, rhAPC had no antibacterial effect. E. coli caused cytokine production to increase many fold. This increase was significantly greater with antibiotics (P < 0.01). Without antibiotics, rhAPC decreased production of TNF-alpha, IL-1beta and IL-6, but not IL-8. At high levofloxacin concentrations, rhAPC was associated with further increases in the concentrations of these cytokines. Cytokine concentrations at 24 h were unaffected by rhAPC in the presence of ampicillin and E. coli. CONCLUSIONS: rhAPC can affect the bactericidal activity and cytokine production of human MDM in the presence of infection and antibiotic therapy. Importantly, factors such as type and concentration of antibiotics, presence of bacteria and timing must be taken into consideration when evaluating cytokine data from septic patients. PMID- 17428811 TI - Reduced mitochondrial density in the vastus lateralis muscle of patients with COPD. AB - Skeletal muscle dysfunction is a well-recognised hallmark of chronic obstructive pulmonary disease (COPD) leading to exercise intolerance. The vastus lateralis of COPD patients is characterised by reduced mitochondrial enzyme activity; however, this is not the case in the tibialis anterior. It is, however, unclear whether the compromised oxidative capacity in the vastus is due to reduced mitochondrial volume density. Muscle biopsies were obtained from the vastus lateralis of six COPD patients and four healthy age-matched controls, and from the tibialis anterior of another six COPD patients and six controls. Mitochondrial number, fractional area and morphometry, as well as Z-line width (as a surrogate marker of fibre type), were analysed using transmission electron microscopy. Mitochondrial number (0.34 versus 0.63 n.microm(-2)) and fractional area (1.95 versus 4.25%) were reduced in the vastus of COPD patients compared with controls. Despite a reduced mitochondrial number (0.65 versus 0.88 n.microm(-2)), the mitochondrial fractional area was maintained in the tibialis of COPD patients compared with controls. It can be concluded that the reduced mitochondrial fractional area is likely to contribute to the decreased oxidative capacity in the vastus of chronic obstructive pulmonary disease patients, whereas the maintained mitochondrial fractional area in the tibialis may explain the normal oxidative capacity. PMID- 17428812 TI - Severity of scleroderma lung disease is related to alveolar concentration of nitric oxide. AB - The alveolar concentration of exhaled nitric oxide (CA,(NO)) is increased in patients with systemic sclerosis (SSc), but whether this increase is related to the severity of interstitial lung disease (ILD) in SSc has not yet been investigated. In total, 58 SSc patients prospectively underwent pulmonary function tests (PFTs), echocardiogram and fibrosis scoring on pulmonary computed tomography (CT). Patients were divided into two groups according to the presence (or not) of ILD. Measurements of CA,(NO) were assessed in all SSc patients and compared with those obtained in 19 healthy volunteers. Relationships were sought between CA,(NO) PFTs and CT scan fibrosis scores. Overall, CA,(NO) was significantly increased in SSc patients (median (range) 6.2 (3.8-9.9) ppb) as compared with controls (2.0 (1.2-3.0) ppb). Among SSc patients, CA,(NO) was significantly higher in patients with ILD compared with patients without ILD (n = 33, 7.5 (5.2-11.9) ppb versus n = 25, 4.9 (3.1-7.0) ppb, respectively). CA,(NO) was inversely related to total lung capacity (r = -0.34) and the diffusing capacity of the lung for carbon monoxide (r = -0.37) and was directly related to CT scan fibrosis scores (r = 0.36). An increased alveolar concentration of exhaled nitric oxide could, at least in part, either reflect or contribute to the severity of lung disease and could be used to noninvasively assess the extent of interstitial lung disease in systemic sclerosis. PMID- 17428813 TI - Beta2-Adrenoceptor polymorphisms and asthma phenotypes: interactions with passive smoking. AB - The aim of the present study was to assess the effects of possible interactions between beta(2)-adrenoceptor gene polymorphisms and passive smoking on forced expiratory volume in one second (FEV(1)), forced vital capacity (FVC) and exhaled nitric oxide (eNO) in children aged 11 yrs. A cross-sectional analysis of the longitudinal cohort was conducted for associations between beta(2)-adrenoceptor gene polymorphisms and lung function and eNO with regard to passive smoking. Among children exposed to tobacco smoke, those with Arg16 (at least one Arg allele) exhibited lower adjusted mean FEV(1) (2.19 versus 2.38 L) and FVC (2.43 versus 2.64 L) than Gly16 homozygotes. Those with Gln27 (at least one Gln allele) also exhibited a lower adjusted mean FEV(1) relative to Glu27 homozygotes (2.24 versus 2.39 L). Among children with no exposure to smoking, those with Arg16 or Gln27 showed lower adjusted geometric mean eNO levels compared with Gly16 homozygotes (15.4 versus 30.9 ppb) and Glu27 homozygotes (18.0 versus 49.7 ppb). In conclusion, passive smoking had a significant effect on associations between beta(2)-adrenoceptor gene polymorphisms and asthma-related phenotypes, enhancing the relationship between Arg16 and lung function and removing the relationship between Arg16 or Gln27 and exhaled nitric oxide levels. PMID- 17428814 TI - Store-refilling involves both L-type calcium channels and reverse-mode sodium calcium exchange in airway smooth muscle. AB - The aim of the present study was to examine the relative contributions to store refilling of the two following voltage-regulated calcium ion influx pathways: 1) L-type-Ca(2+) channels; and 2) the reverse-mode of the sodium-calcium exchanger (NCX). Successive acetylcholine-induced contractions, triggered in bovine tracheal smooth muscle strips, were measured to determine the effect of intervention on contractions as an indication of the extent of store-refilling. Pre-treating the tissues with cromakalim significantly reduced the magnitude of successive contractions. Zero-Ca(2+) bathing media abolished the contractions, an effect that was completely reversed upon reintroduction of Ca(2+). Inhibition of L-type Ca(2+) channels, with nifedipine, significantly reduced the magnitude of the contractions. Similarly, inhibition of the reverse-mode of the NCX, with KB R7943, significantly reduced the magnitude of the contractions. However, neither nifedipine nor KB-R7943 alone reduced contractions to the same extent as observed under zero-Ca(2+) conditions. Concurrent treatment with nifedipine and KB-R7943 almost abolished successive contractions. Furthermore, concurrent treatment with nifedipine and zero-Na(+) bathing media displayed a significantly greater effect than nifedipine alone. Probing the expression of NCX1 isoforms by Western blotting revealed the presence of three bands of 160, 120 and 110 kDa. The 120- and 110-kDa bands were identified as variably spliced NCX isoforms, NCX1.1 and NCX1.3, respectively. Taken together, these data suggest that influx of calcium ions through both L-type calcium channels and the reverse-mode of the sodium calcium exchanger is necessary for complete store-refilling in airway smooth muscle. PMID- 17428815 TI - An integrative genomics strategy for systematic characterization of genetic loci modulating phenotypes. AB - Naturally occurring genetic variations may affect certain phenotypes through influencing transcript levels of the genes that are causally related to those phenotypes. Genomic regions harboring common sequence variants that modulate gene expression can be mapped as quantitative trait loci (QTLs) using a newly developed genetical genomics approach. This enables a new strategy for systematically mapping novel genetic loci underlying various phenotypes. In this work, we started from a seed set of genes with variants that are known to affect behavioral and neurological phenotypes (as recorded in Mammalian Phenotype Ontology Database) and used microarrays to analyze their expression levels in brain samples of a panel of BXD recombinant inbred mouse strains. We then systematically mapped the QTLs controlling the expression of these genes. Candidate causal genes in the QTL intervals were evaluated for evidence of functional genetic polymorphisms. Using this method, we were able to predict novel genetic loci and causal genes for a number of behavioral and neurological phenotypes. Lines of independent evidence supporting some of our results were provided by transcription factor binding site analysis and by biomedical literature. This strategy integrates gene-phenotype relations from decades of experimental mutagenesis studies and new genomic resources to provide an approach to rapidly expand knowledge on genetic loci modulating phenotypes. PMID- 17428816 TI - Opa1 deficiency in a mouse model of autosomal dominant optic atrophy impairs mitochondrial morphology, optic nerve structure and visual function. AB - OPA1 is a ubiquitously expressed, nuclear dynamin-related GTPase, targeted to the inner mitochondrial membrane, which plays a role in mitochondrial fusion. Mutations in the OPA1 gene on chromosome 3q28-qter are associated with autosomal dominant optic atrophy (ADOA), the most common inherited optic neuropathy, in which retinal ganglion cells (RGCs) are lost and visual acuity is impaired from an early age. We have generated a novel ENU-induced mutant mouse carrying a protein-truncating nonsense mutation in opa1 in order to explore the pathophysiology of ADOA. The heterozygous mutation, B6; C3-Opa1(Q285STOP), located in exon 8 immediately before the central dynamin-GTPase, leads to approximately 50% reduction in opa1 protein in retina and all tissues on western analysis. The homozygous mutation is embryonic lethal by 13.5 days post coitum, demonstrating the importance of Opa1 during early development. Fibroblasts taken from adult heterozygous mutant mice show an apparent alteration in morphology, with an increase in mitochondrial fission and fragmentation. Heterozygous mutants show a slow onset of degeneration in the optic nerve electron microscopy. Furthermore, they demonstrate a functional reduction in visual function on testing with the optokinetic drum and the circadian running wheel. These findings indicate that the opa1 GTPase contains crucial information required for the survival of RGCs and that Opa1 is essential for early embryonic survival. The Opa1 +/- mice described here provide a means to directly investigate the cellular pathophysiology of OPA1 ADOA. PMID- 17428817 TI - Decision making in asthma exacerbation: a clinical judgement analysis. AB - BACKGROUND: Clinical decisions which impact directly on patient safety and quality of care are made during acute asthma attacks by individual doctors based on their knowledge and experience. Decisions include administration of systemic corticosteroids (CS) and oral antibiotics, and admission to hospital. Clinical judgement analysis provides a methodology for comparing decisions between practitioners with different training and experience, and improving decision making. METHODS: Stepwise linear regression was used to select clinical cues based on visual analogue scale assessments of the propensity of 62 clinicians to prescribe a short course of oral CS (decision 1), a course of antibiotics (decision 2), and/or admit to hospital (decision 3) for 60 "paper" patients. RESULTS: When compared by specialty, paediatricians' models for decision 1 were more likely to include level of alertness as a cue (54% vs 16%); for decision 2 they were more likely to include presence of crepitations (49% vs 16%) and less likely to include inhaled CS (8% vs 40%), respiratory rate (0% vs 24%) and air entry (70% vs 100%). When compared to other grades, the models derived for decision 3 by consultants/general practitioners were more likely to include wheeze severity as a cue (39% vs 6%). CONCLUSIONS: Clinicians differed in their use of individual cues and the number included in their models. Patient safety and quality of care will benefit from clarification of decision-making strategies as general learning points during medical training, in the development of guidelines and care pathways, and by clinicians developing self-awareness of their own preferences. PMID- 17428818 TI - Computerised paediatric asthma quality of life questionnaires in routine care. AB - BACKGROUND: Asthma quality of life questionnaires are not readily incorporated into clinical care. We therefore computerised the Paediatric Asthma Quality of Life Questionnaire (standardised) (PAQLQ(S)) and the Paediatric Asthma Caregivers Quality of Life Questionnaire (PACQLQ), with a colour-coded printed graphical report. OBJECTIVES: To (a) assess the feasibility of the electronic questionnaires in clinical care and (b) compare the child's PAQLQ scores with the parent's score, physician's clinical score and spirometry. METHODS: Children with asthma were given a clinical severity score of 1-4 (increasing severity) and then completed the PAQLQ(S) electronically (scores 1-7 for increasing quality of life in emotional, symptoms and activity limitation domains) followed by spirometry and physician review. Parents completed the PACQLQ. Inclusion criteria required fluent Hebrew and reliable performance of spirometry. Children with additional chronic diseases were excluded. RESULTS: 147 children with asthma aged 7-17 years completed PAQLQs and 115 accompanying parents completed PACQLQs, taking 8.3 (4.3 15) and 4.4 (1.5-12.7) min, respectively (mean (range)). Graphical reports enabled physicians to address quality of life during even brief visits. Children's (PAQLQ) and parents' (PACQLQ) total scores correlated (r = 0.61, p<0.001), although the children's median emotional score of 6.3 was higher than their parents' 5.7 (p<0.001), whereas median activity limitation score was lower than their parents': 5.0 and 6.8, respectively (p<0.001). No correlation was found with physician's clinical score or spirometry. CONCLUSIONS: Electronic PAQLQs are easy to use, providing additional insight to spirometry and physician's assessment, in routine asthma care. Future studies must assess impact on asthma management. PMID- 17428819 TI - Prevalence and pathogenesis of congenital anomalies in cerebral palsy. AB - BACKGROUND: It has been hypothesised that cerebral palsy (CP) and other congenital anomalies are attributable to feto-fetal transfusion problems in a monochorionic multiple gestation. Thus more than one organ could be compromised leading to the coexistence of two or more anomalies in a fetus. Such anomalies in a singleton birth may be attributable to early demise of the co-conceptus as a vanishing twin. AIM: To determine whether the coexistence of congenital anomalies and CP is greater than a chance finding by comparing the prevalence of congenital anomalies in children with CP with that in the general population of children. METHODS: A population-based register of children with CP born in 1966-1991 in the counties of Merseyside and Cheshire, UK, comprised the index population. Coexisting congenital anomalies were recorded. For comparison the population prevalence of congenital anomalies was obtained from eight congenital malformation registers in the UK. RESULTS: Children with CP were found to have highly significant increases in risk for microcephaly, isolated hydrocephaly, congenital anomalies of the eye, congenital cardiac anomalies, cleft lip and/or palate and congenital dislocation of the hips and talipes (p<0.001) and atresias of the oesophagus (p<0.001) and intestines (p<0.01). The relative risks ranged from 3.1 (95% CI 1.9 to 4.8; p<0.001) for congenital malformations of the cardiac septa to 116.09 (95% CI 84.0 to 162.3; p<0.001) for microcephaly. CONCLUSIONS: Congenital anomalies in children with CP are found much more frequently than expected by chance. A common pathogenic mechanism may account for the coexistence of disparate congenital anomalies. A hypothesis is proposed for such a common pathogenic mechanism. PMID- 17428820 TI - Consequential alterations in haemoglobin structure upon glycation with fructose: prevention by acetylsalicylic acid. AB - Increased fructose concentration in erythrocytes of diabetic patients subject haemoglobin (Hb) to be glycated by fructose. Haemoglobin glycation results in early and advanced glycation end products which are known as HbA(1c) and Hb-AGE, respectively. In diabetics the Hb-AGE content raises and the mean parameter multiplication value (PMV) for Hb-AGE has been calculated at 6 against non diabetics. We are emphasizing on fluorescence descriptive pairs especially of 370/440 nm (lambda(ex)/lambda(em)) and thioflavin T binding process to characterize short-term Hb-AGE formation upon glycation with fructose or fructation. Secondary structure elements were estimated after far-UV circular dichroism spectropolarimetery which reveals up to 11.3% beta-content attainment for 20 days incubated Hb with fructose which is concurred with enhanced hydrophobicity of the protein. The later is described through enhanced 1-anilino 8-naphthalene sulphonate (ANS) binding to fructated protein. The preventive effect of acetylsalicylic acid on glycation of methemoglobin and consequently, Hb AGE formation also has been included. PMID- 17428821 TI - Contribution of the metabolic syndrome to sudden death risk in asymptomatic men: the Paris Prospective Study I. AB - AIMS: To compare the risk of sudden death and non-sudden death from myocardial infarction associated with the metabolic syndrome (MetS) in asymptomatic men. METHODS AND RESULTS: The mortality status of 6,678 middle-aged men from the Paris Prospective Study I, who were free of diabetes and coronary heart disease (CHD) at the baseline examination, has been investigated over 21 years of follow-up. The sagittal abdominal diameter was substituted for waist circumference, and HDL cholesterol was unavailable. The presence of three abnormalities and the presence of abdominal adiposity plus at least two abnormalities defined the MetS, using the NCEP-ATP III and IDF criteria, respectively. Frequency estimate of the MetS was 14.4 and 16.7%, using the NCEP-ATP III and IDF criteria, respectively. The MetS increased the risk of sudden death and non-sudden death by 68% [95% confidence interval (CI) 1.05-2.70] and 38% (95% CI 0.95-2.01), respectively, after adjustment for other CHD risk factors (P for the comparison of the hazard ratios = 0.25). Hazards ratio using the IDF criteria were 2.02 (95% CI 1.30-3.14) and 1.69 (95% CI 1.20-2.38), respectively, (P = 0.26). CONCLUSION: In healthy middle-aged men, the MetS increased the risk of sudden death and, to a lesser extent, the risk of non-sudden death over 21 years independent of CHD risk factors. PMID- 17428822 TI - How to diagnose diastolic heart failure: a consensus statement on the diagnosis of heart failure with normal left ventricular ejection fraction by the Heart Failure and Echocardiography Associations of the European Society of Cardiology. AB - Diastolic heart failure (DHF) currently accounts for more than 50% of all heart failure patients. DHF is also referred to as heart failure with normal left ventricular (LV) ejection fraction (HFNEF) to indicate that HFNEF could be a precursor of heart failure with reduced LVEF. Because of improved cardiac imaging and because of widespread clinical use of plasma levels of natriuretic peptides, diagnostic criteria for HFNEF needed to be updated. The diagnosis of HFNEF requires the following conditions to be satisfied: (i) signs or symptoms of heart failure; (ii) normal or mildly abnormal systolic LV function; (iii) evidence of diastolic LV dysfunction. Normal or mildly abnormal systolic LV function implies both an LVEF > 50% and an LV end-diastolic volume index (LVEDVI) <97 mL/m(2). Diagnostic evidence of diastolic LV dysfunction can be obtained invasively (LV end-diastolic pressure >16 mmHg or mean pulmonary capillary wedge pressure >12 mmHg) or non-invasively by tissue Doppler (TD) (E/E' > 15). If TD yields an E/E' ratio suggestive of diastolic LV dysfunction (15 > E/E' > 8), additional non invasive investigations are required for diagnostic evidence of diastolic LV dysfunction. These can consist of blood flow Doppler of mitral valve or pulmonary veins, echo measures of LV mass index or left atrial volume index, electrocardiographic evidence of atrial fibrillation, or plasma levels of natriuretic peptides. If plasma levels of natriuretic peptides are elevated, diagnostic evidence of diastolic LV dysfunction also requires additional non invasive investigations such as TD, blood flow Doppler of mitral valve or pulmonary veins, echo measures of LV mass index or left atrial volume index, or electrocardiographic evidence of atrial fibrillation. A similar strategy with focus on a high negative predictive value of successive investigations is proposed for the exclusion of HFNEF in patients with breathlessness and no signs of congestion. The updated strategies for the diagnosis and exclusion of HFNEF are useful not only for individual patient management but also for patient recruitment in future clinical trials exploring therapies for HFNEF. PMID- 17428823 TI - Wiring the nervous system: from form to function. AB - The RIKEN Centre for Developmental Biology recently hosted a joint UK-Asian Pacific Developmental Biology Network meeting called 'Development and Emergence of Function in the Nervous System'. The meeting's program, which was organized by James Briscoe and Krishnaswamy VijayRaghavan, covered a spectrum of processes and mechanisms in neurodevelopment, ranging from the patterning of neural tissue to the initiation of a functional nervous system. One idea to have emerged during this meeting is that 'form underlies function'. Here we discuss some of the themes that were addressed and provide a broad impression of what was a highly stimulating and successful conference. PMID- 17428824 TI - Nab controls the activity of the zinc-finger transcription factors Squeeze and Rotund in Drosophila development. AB - Nab proteins form an evolutionarily conserved family of transcriptional co regulators implicated in multiple developmental events in various organisms. They lack DNA-binding domains and act by associating with other transcription factors, but their precise roles in development are not known. Here we analyze the role of nab in Drosophila development. By employing genetic approaches we found that nab is required for proximodistal patterning of the wing imaginal disc and also for determining specific neuronal fates in the embryonic CNS. We identified two partners of Nab: the zinc-finger transcription factors Rotund and Squeeze. Nab is co-expressed with squeeze in a subset of neurons in the embryonic ventral nerve cord and with rotund in a circular domain of the distal-most area of the wing disc. Our results indicate that Nab is a co-activator of Squeeze and is required to limit the number of neurons that express the LIM-homeodomain gene apterous and to specify Tv neuronal fate. Conversely, Nab is a co-repressor of Rotund in wing development and is required to limit the expression of wingless (wg) in the wing hinge, where wg plays a mitogenic role. We also showed by pull-down assays that Nab binds directly to Rotund and Squeeze via its conserved C-terminal domain. We propose two mechanisms by which the activation of wg expression by Rotund in the wing hinge is repressed in the distal wing. PMID- 17428825 TI - Specification of Arabidopsis floral meristem identity by repression of flowering time genes. AB - Flowering plants produce floral meristems in response to intrinsic and extrinsic flowering inductive signals. In Arabidopsis, the floral meristem identity genes LEAFY (LFY) and APETALA1 (AP1) are activated to play a pivotal role in specifying floral meristems during floral transition. We show here that the emerging floral meristems require AP1 to partly specify their floral identities by directly repressing a group of flowering time genes, including SHORT VEGETATIVE PHASE (SVP), AGAMOUS-LIKE 24 (AGL24) and SUPPRESSOR OF OVEREXPRESSION OF CO1 (SOC1). In wild-type plants, these flowering time genes are normally downregulated in emerging floral meristems. In the absence of AP1, these genes are ectopically expressed, transforming floral meristems into shoot meristems. By post translational activation of an AP1-GR fusion protein and chromatin immunoprecipitation assays, we further demonstrate the repression of these flowering time genes by induced AP1 activity and in vivo AP1 binding to the cis regulatory regions of these genes. These findings indicate that once AP1 is activated during the floral transition, it acts partly as a master repressor in floral meristems by directly suppressing the expression of flowering time genes, thus preventing the continuation of the shoot developmental program. PMID- 17428826 TI - Akt mediates self-renewal division of mouse spermatogonial stem cells. AB - Spermatogonial stem cells have unique properties to self-renew and support spermatogenesis throughout their lifespan. Although glial cell line-derived neurotrophic factor (GDNF) has recently been identified as a self-renewal factor for spermatogonial stem cells, the molecular mechanism of spermatogonial stem cell self-renewal remains unclear. In the present study, we assessed the role of the phosphoinositide-3 kinase (PI3K)-Akt pathway using a germline stem (GS) cell culture system that allows in vitro expansion of spermatogonial stem cells. Akt was rapidly phosphorylated when GDNF was added to the GS cell culture, and the addition of a chemical inhibitor of PI3K prevented GS cell self-renewal. Furthermore, conditional activation of the myristoylated form of Akt-Mer (myr-Akt Mer) by 4-hydroxy-tamoxifen induced logarithmic proliferation of GS cells in the absence of GDNF for at least 5 months. The myr-Akt-Mer GS cells expressed spermatogonial markers and retained androgenetic imprinting patterns. In addition, they supported spermatogenesis and generated offspring following spermatogonial transplantation into the testes of infertile recipient mice, indicating that they are functionally normal. These results demonstrate that activation of the PI3K-Akt pathway plays a central role in the self-renewal division of spermatogonial stem cells. PMID- 17428827 TI - Dynamic decapentaplegic signaling regulates patterning and adhesion in the Drosophila pupal retina. AB - The correct organization of cells within an epithelium is essential for proper tissue and organ morphogenesis. The role of Decapentaplegic/Bone morphogenetic protein (Dpp/BMP) signaling in cellular morphogenesis during epithelial development is poorly understood. In this paper, we used the developing Drosophila pupal retina--looking specifically at the reorganization of glial-like support cells that lie between the retinal ommatidia--to better understand the role of Dpp signaling during epithelial patterning. Our results indicate that Dpp pathway activity is tightly regulated across time in the pupal retina and that epithelial cells in this tissue require Dpp signaling to achieve their correct shape and position within the ommatidial hexagon. These results point to the Dpp pathway as a third component and functional link between two adhesion systems, Hibris-Roughest and DE-cadherin. A balanced interplay between these three systems is essential for epithelial patterning during morphogenesis of the pupal retina. Importantly, we identify a similar functional connection between Dpp activity and DE-cadherin and Rho1 during cell fate determination in the wing, suggesting a broader link between Dpp function and junctional integrity during epithelial development. PMID- 17428828 TI - A crucial role for Olig2 in white matter astrocyte development. AB - The mechanisms underlying astrocyte heterogeneity in the developing mouse brain are poorly understood. The bHLH transcription factor Olig2 is essential for motoneuron and oligodendrocyte formation; however, its role in astrocyte development remains obscure. During cortical development, Olig2 is transiently expressed in immature developing astrocytes at neonatal stages and is progressively downregulated in astrocytes at late postnatal stages. To assess the function of Olig2 in astrocyte formation, we conditionally ablated Olig2 in a spatiotemporally controlled manner. In the Olig2-ablated cortex and spinal cord, the formation of astrocytes in the white matter is severely compromised. Temporally controlled mutagenesis revealed that postnatal Olig2 function is required for astrocyte differentiation in the cerebral white matter. By contrast, astrocytes in the cortical gray matter are formed, but with sustained GFAP upregulation in the superficial layers. Cell type-specific mutagenesis and fate mapping analyses indicate that abnormal astrocyte formation is at least in part attributable to the loss of Olig2 in developing astrocytes and their precursors. Thus, our studies uncover a crucial role for Olig2 in white matter astrocyte development and reveal divergent transcriptional requirements for, and developmental sources of, morphologically and spatially distinct astrocyte subpopulations. PMID- 17428829 TI - Foxp2 and Foxp1 cooperatively regulate lung and esophagus development. AB - The airways of the lung develop through a reiterative process of branching morphogenesis that gives rise to the intricate and extensive surface area required for postnatal respiration. The forkhead transcription factors Foxp2 and Foxp1 are expressed in multiple foregut-derived tissues including the lung and intestine. In this report, we show that loss of Foxp2 in mouse leads to defective postnatal lung alveolarization, contributing to postnatal lethality. Using in vitro and in vivo assays, we show that T1alpha, a lung alveolar epithelial type 1 cell-restricted gene crucial for lung development and function, is a direct target of Foxp2 and Foxp1. Remarkably, loss of a single Foxp1 allele in addition to complete loss of Foxp2 results in increased severity of morphological defects in mutant lungs and leads to perinatal loss of all Foxp2(-/-);Foxp1(+/-) mice. Expression of N-myc and Hop, crucial regulators of lung development, is compromised in Foxp2(-/-);Foxp1(+/-) mutants. In addition to the defects in lung development, esophageal muscle development is disrupted in Foxp2(-/-);Foxp1(+/-) embryos, a tissue where Foxp2 and Foxp1 are co-expressed. These data identify Foxp2 and Foxp1 as crucial regulators of lung and esophageal development, underscoring the necessity of these transcription factors in the development of anterior foregut-derived tissues and demonstrating functional cooperativity between members of the Foxp1/2/4 family in tissues where they are co-expressed. PMID- 17428830 TI - Selective requirements for NRP1 ligands during neurovascular patterning. AB - Blood vessels and neurons share several types of guidance cues and cell surface receptors to control their behaviour during embryogenesis. The transmembrane protein NRP1 is present on blood vessels and nerves. NRP1 binds two structurally diverse ligands, the semaphorin SEMA3A and the VEGF164 isoform of vascular endothelial growth factor. SEMA3A was originally identified as a repulsive cue for developing axons that acts by signalling through receptor complexes containing NRP1 and plexins. In vitro, SEMA3A also inhibits integrin function and competes with VEGF164 for binding to NRP1 to modulate the migration of endothelial cells. These observations resulted in a widely accepted model of vascular patterning in which the balance of VEGF164 and SEMA3A determines endothelial cell behaviour. However, we now demonstrate that SEMA3A is not required for angiogenesis in the mouse, which instead is controlled by VEGF164. We find that SEMA3A, but not VEGF164, is required for axon patterning of limb nerves, even though the competition between VEGF164 and SEMA3A for NRP1 affects the migration of neuronal progenitor cells in vitro and has been hypothesised to control axon guidance. Moreover, we show that there is no genetic interaction between SEMA3A and VEGF164 during vasculogenesis, angiogenesis or limb axon patterning, suggesting that ligand competition for NRP1 binding cannot explain neurovascular congruence, as previously suggested. We conclude that NRP1 contributes to both neuronal and vascular patterning by preferentially relaying SEMA3A signals in peripheral axons and VEGF164 signals in blood vessels. PMID- 17428831 TI - pygopus 2 has a crucial, Wnt pathway-independent function in lens induction. AB - Drosophila Pygopus was originally identified as a core component of the canonical Wnt signaling pathway and a transcriptional coactivator. Here we have investigated the microophthalmia that arises in mice with a germline null mutation of pygopus 2. We show that this phenotype is a consequence of defective lens development at inductive stages. Using a series of regionally limited Cre recombinase transgenes for conditional deletion of Pygo2(flox), we show that Pygo2 activity in pre-placodal presumptive lens ectoderm, placodal ectoderm and ocular mesenchyme all contribute to lens development. In each case, Pygo2 is required for normal expression levels of the crucial transcription factor Pax6. Finally, we provide multiple lines of evidence that although Pygo2 can function in the Wnt pathway, its activity in lens development is Wnt pathway-independent. PMID- 17428832 TI - Responses of photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice. AB - BACKGROUND AND AIMS: Salinity is a widespread soil problem limiting productivity of cereal crops worldwide. Rice is particularly sensitive to salt stress during the seedling stage, with consequent poor crop establishment, as well as during reproduction where salinity can severely disrupt grain formation and yield. Tolerance at the seedling stage is weakly associated with tolerance during reproduction. Physiological responses to salinity were evaluated for contrasting genotypes, during the seedling and reproductive stages. METHODS: Three rice genotypes differing in their tolerance of salinity were evaluated in a set of greenhouse experiments under salt stress during both seedling stage and reproduction. KEY RESULTS: Photosynthetic CO2 fixation, stomatal conductance (gs) and transpiration decreased substantially because of salt stress, but with greater reduction in the sensitive cultivar IR29. The tolerant lines IR651 and IR632 had more responsive stomata that tended to close faster during the first few hours of stress, followed by partial recovery after a brief period of acclimation. However, in the sensitive line, gs continued to decrease for longer duration and with no recovery afterward. Chlorophyll fluorescence measurements revealed that non-photochemical quenching increased, whereas the electron transport rate decreased under salt stress. Salt-tolerant cultivars exhibited much lower lipid peroxidation, maintained elevated levels of reduced ascorbic acid and showed increased activities of the enzymes involved in the reactive oxygen scavenging system during both developmental stages. CONCLUSIONS: Upregulation of the anti-oxidant system appears to play a role in salt tolerance of rice, with tolerant genotypes also maintaining relatively higher photosynthetic function; during both the vegetative and reproductive stages. PMID- 17428833 TI - Responses of rice cultivars with different nitrogen use efficiency to partial nitrate nutrition. AB - BACKGROUND AND AIMS: There is increased evidence that partial nitrate (NO3-) nutrition (PNN) improves growth of rice (Oryza sativa), although the crop prefers ammonium (NH4+) to NO3- nutrition. It is not known whether the response to NO3- supply is related to nitrogen (N) use efficiency (NUE) in rice cultivars. Methods Solution culture experiments were carried out to study the response of two rice cultivars, Nanguang (High-NUE) and Elio (Low-NUE), to partial NO3- supply in terms of dry weight, N accumulation, grain yield, NH4+ uptake and ammonium transporter expression [real-time polymerase chain reaction (PCR)]. KEY RESULTS: A ratio of 75/25 NH4+ -N/NO3- -N increased dry weight, N accumulation and grain yield of 'Nanguang' by 30, 36 and 21 %, respectively, but no effect was found in 'Elio' when compared with those of 100/0 NH4+ -N/NO3- -N. Uptake experiments with 15N-NH4+ showed that NO3- increased NH4+ uptake efficiency in 'Nanguang' by increasing Vmax (14 %), but there was no effect on Km. This indicated that partial replacement of NH4+ by NO3- could increase the number of the ammonium transporters but did not affect the affinity of the transporters for NH4+. Real time PCR showed that expression of OsAMT1s in 'Nanguang' was improved by PNN, while that in 'Elio' did not change, which is in accordance with the differing responses of these two cultivars to PNN. Conclusions Increased NUE by PNN can be attributed to improved N uptake. The rice cultivar with a higher NUE has a more positive response to PNN than that with a low NUE, suggesting that there might be a relationship between PNN and NUE. PMID- 17428834 TI - Effect of temperature, light and salinity on seed germination and radicle growth of the geographically widespread halophyte shrub Halocnemum strobilaceum. AB - BACKGROUND AND AIMS: The small leafy succulent shrub Halocnemum strobilaceum occurs in saline habitats from northern Africa and Mediterranean Europe to western Asia, and it is a dominant species in salt deserts such as those of north west China. The effects of temperature, light/darkness and NaCl salinity were tested on seed germination, and the effects of salinity were tested on seed germination recovery, radicle growth and radicle elongation recovery, using seeds from north-west China; the results were compared with those previously reported on this species from 'salt steppes' in the Mediterranean region of Spain. METHODS: Seed germination was tested over a range of temperatures in light and in darkness and over a range of salinities at 25 degrees C in the light. Seeds that did not germinate in the NaCl solutions were tested for germination in deionized water. Seeds from which radicles had barely emerged in deionized water were transferred to NaCl solutions for 10 d and then back to deionized water for 10 d to test for radicle growth and recovery. KEY RESULTS: Seeds germinated to higher percentages in light than in darkness and at high than at low temperatures. Germination percentages decreased with an increase in salinity from 0.1 to 0.75 M NaCl. Seeds that did not germinate in NaCl solutions did so after transfer to deionized water. Radicle elongation was increased by low salinity, and then it decreased with an increase in salinity, being completely inhibited by > or = 2.0 M NaCl. Elongation of radicles from salt solutions < 3.0 M resumed after seedlings were transferred to deionized water. CONCLUSIONS: The seed and early seedling growth stages of the life cycle of H. strobilaceum are very salt tolerant, and their physiological responses differ somewhat between the Mediterranean 'salt steppe' of Spain and the inland cold salt desert of north west China. PMID- 17428835 TI - Evolution of perianth and stamen characteristics with respect to floral symmetry in Ranunculales. AB - BACKGROUND AND AIMS: Floral symmetry presents two main states in angiosperms, namely polysymmetry and monosymmetry. Monosymmetry is thought to have evolved several times independently from polysymmetry, possibly in co-adaptation with specialized pollinators. Monosymmetry commonly refers to the perianth, even though associated androecium modifications have been reported. The evolution of perianth symmetry is examined with respect to traits of flower architecture in the Ranunculales, the sister group to all other eudicots, which present a large diversity of floral forms. METHODS: Characters considered were perianth merism, calyx, corolla and androecium symmetry, number of stamens and spurs. Character evolution was optimized on a composite phylogenetic tree of Ranunculales using maximum parsimony. KEY RESULTS: The ancestral state for merism could not be inferred because the basalmost Eupteleaceae lack a perianth and have a variable number of stamens. The Papaveraceae are dimerous, and the five other families share a common trimerous ancestor. Shifts from trimery to dimery (or reverse) are observed. Pentamery evolved in Ranunculaceae. Ranunculales except Eupteleaceae, present a polysymmetric ancestral state. Monosymmetry evolved once within Papaveraceae, Ranunculaceae and Menispermaceae (female flowers only). Oligandry is the ancestral state for all Ranunculales, and polyandry evolved several times independently, in Papaveraceae, Menispermaceae, Berberidaceae and Ranunculaceae, with two reversions to oligandry in the latter. The ancestral state for androecium symmetry is ambiguous for the Ranunculales, while polysymmetry evolved immediately after the divergence of Eupteleaceae. A disymmetric androecium evolved in Papaveraceae. The ancestral state for spurs is none. Multiple spurs evolved in Papaveraceae, Berberidaceae and Ranunculaceae, and single spurs occur in Papaveraceae and Ranunculaceae. CONCLUSIONS: The evolution of symmetry appears disconnected from changes in merism and stamen number, although monosymmetry never evolved in the context of an open ground plan. In bisexual species, monosymmetry evolved coincidently with single spurs, allowing us to propose an evolutionary scenario for Papaveraceae. PMID- 17428836 TI - Aberrant hemichannel properties of Cx26 mutations causing skin disease and deafness. AB - Mutations in the human GJB2 gene, which encodes connexin26 (Cx26), underlie various forms of hereditary deafness and skin disease. While it has proven difficult to discern the exact pathological mechanisms that cause these disorders, studies have shown that the loss or abnormal function of Cx26 protein has a profound effect on tissue homeostasis. Here, we used the Xenopus oocyte expression system to examine the functional characteristics of a Cx26 mutation (G45E) that results in keratitis-ichthyosis-deafness syndrome (KIDS) with a fatal outcome. Our data showed that oocytes were able to express both wild-type Cx26 and its G45E variant, each of which formed hemichannels and gap junction channels. However, Cx26-G45E hemichannels displayed significantly greater whole cell currents than wild-type Cx26, leading to cell lysis and death. This severe phenotype could be rescued in the presence of elevated Ca(2+) levels in the extracellular milieu. Cx26-G45E could also form intercellular channels with a similar efficiency as wild-type Cx26, however, with increased voltage sensitive gating. We also compared Cx26-G45E with a previously described Cx26 mutant, A40V, which has an overlapping human phenotype. We found that both dominant Cx26 mutants elicited similar functional consequences and that cells coexpressing mutant and wild-type connexins predominantly displayed mutant-like behavior. These data suggest that mutant hemichannels may act on cellular homeostasis in a manner that can be detrimental to the tissues in which they are expressed. PMID- 17428837 TI - Adverse effects of the classic antioxidant uric acid in adipocytes: NADPH oxidase mediated oxidative/nitrosative stress. AB - Uric acid is considered a major antioxidant in human blood that may protect against aging and oxidative stress. Despite its proposed protective properties, elevated levels of uric acid are commonly associated with increased risk for cardiovascular disease and mortality. Furthermore, recent experimental studies suggest that uric acid may have a causal role in hypertension and metabolic syndrome. All these conditions are thought to be mediated by oxidative stress. In this study we demonstrate that differentiation of cultured mouse adipocytes is associated with increased production of reactive oxygen species (ROS) and uptake of uric acid. Soluble uric acid stimulated an increase in NADPH oxidase activity and ROS production in mature adipocytes but not in preadipocytes. The stimulation of NADPH oxidase-dependent ROS by uric acid resulted in activation of MAP kinases p38 and ERK1/2, a decrease in nitric oxide bioavailability, and an increase in protein nitrosylation and lipid oxidation. Collectively, our results suggest that hyperuricemia induces redox-dependent signaling and oxidative stress in adipocytes. Since oxidative stress in the adipose tissue has recently been recognized as a major cause of insulin resistance and cardiovascular disease, hyperuricemia-induced alterations in oxidative homeostasis in the adipose tissue might play an important role in these derangements. PMID- 17428838 TI - Viscoelasticity of the human red blood cell. AB - We report here the first measurements of the complex modulus of the isolated red blood cell (RBC). Because the RBC is often larger than capillary diameter, important determinants of microcirculatory function are RBC deformability and its changes with pathologies, such as sickle cell disease and malaria. A functionalized ferrimagnetic microbead was attached to the membrane of healthy RBC and then subjected to an oscillatory magnetic field. The resulting torque caused cell deformation. From the oscillatory forcing and resulting bead motions, which were tracked optically, we computed elastic and frictional moduli, g' and g", respectively, from 0.1 to 100 Hz. The g' was nearly frequency independent and dominated the response at all but the highest frequencies measured. Over three frequency decades, g" increased as a power law with an exponent of 0.64, a result not predicted by any simple model. These data suggest that RBC relaxation times that have been reported previously, and any models that rest upon them, are artifactual; the artifact, we suggest, arises from forcing to an exponential fit data of limited temporal duration. A linear range of response was observed, but, as forcing amplitude increased, nonlinearities became clearly apparent. A finite element model suggests that membrane bending was localized to the vicinity of the bead and dominated membrane shear. While the mechanisms accounting for these RBC dynamics remain unclear, methods described here establish new avenues for the exploration of connections among the mechanical, chemical, and biological characteristics of the RBC in health and disease. PMID- 17428840 TI - Migration of leukocytes across an endothelium-epithelium bilayer as a model of renal interstitial inflammation. AB - Interstitial inflammation has emerged as a key event in the development of acute renal failure. To gain better insight into the nature of these inflammatory processes, the interplay between tubular epithelial cells, endothelial cells, and neutrophils (PMN) was investigated. A coculture transmigration model was developed, composed of human dermal microvascular endothelial (HDMEC) and human renal proximal tubular cells (HK-2) cultured on opposite sides of Transwell growth supports. Correct formation of an endoepithelial bilayer was verified by light and electron microscopy. The model was used to study the effects of endotoxin (LPS), tumor necrosis factor (TNF)-alpha, and alpha-melanocyte stimulating hormone (alpha-MSH) by measuring PMN migration and cytokine release. To distinguish between individual roles of microvascular endothelial and epithelial cells in transmigration processes, migration of PMN was investigated separately in HK-2 and HDMEC monolayers. Sequential migration of PMN through endothelium and epithelium could be observed and was significantly increased after proinflammatory stimulation with either TNF-alpha or LPS (3.5 +/- 0.58 and 2.76 +/- 0.64-fold vs. control, respectively). Coincubation with alpha-MSH inhibited the transmigration of PMN through the bilayer after proinflammatory stimulation with LPS but not after TNF-alpha. The bilayers produced significant amounts of IL-8 and IL-6 mostly released from the epithelial cells. Furthermore, alpha-MSH decreased LPS-induced IL-6 secretion by 30% but had no significant effect on IL-8 secretion. We established a transmigration model showing sequential migration of PMN across microvascular endothelial and renal tubular epithelial cells stimulated by TNF-alpha and LPS. Anti-inflammatory effects of alpha-MSH in this bilayer model are demonstrated by inhibition on PMN transmigration and IL-6 secretion. PMID- 17428839 TI - Selective knockdown of AT1 receptors by RNA interference inhibits Val5-ANG II endocytosis and NHE-3 expression in immortalized rabbit proximal tubule cells. AB - Receptor-mediated endocytosis of extracellular ANG II has been suggested to play an important role in the regulation of proximal tubule cell (PTC) function. Using immortalized rabbit PTCs as an in vitro cell culture model, we tested the hypothesis that extracellular ANG II is taken up by PTCs through angiotensin type 1 receptor (AT(1); or AT(1a)) receptor-mediated endocytosis and that inhibition of ANG II endocytosis using a selective AT(1) receptor small-interfering RNA (siRNA; AT(1)R siRNA) or endocytotic inhibitors exerts a physiological effect on total and apical sodium and hydrogen exchanger isoform 3 (NHE-3) protein abundance. Western blots and live cell imaging with FITC-labeled ANG II confirmed that transfection of PTCs with a human specific AT(1)R siRNA for 48 h selectively knocked down AT(1) receptor protein by 76 +/- 5% (P < 0.01), whereas transfection with a scrambled siRNA had little effect. In nontransfected PTCs, exposure to extracellular ANG II (1 nM) for 60 min at 37 degrees C increased intracellular ANG II accumulation by 67% (control: 566 +/- 55 vs. ANG II: 943 +/- 160 pg/mg protein, P < 0.05) and induced mitogen-activated protein kinase extracellular signal-regulated kinase (ERK) 1/2 phosphorylation (163 +/- 15% of control, P < 0.01). AT(1)R siRNA reduced ANG II endocytosis to a level similar to losartan, which blocks cell surface AT(1) receptors (557 +/- 37 pg/mg protein, P < 0.05 vs. ANG II), or to colchicine, which disrupts cytoskeleton microtubules (613 +/- 12 pg/mg protein, P < 0.05 vs. ANG II). AT(1)R siRNA, losartan, and colchicine all attenuated ANG II-induced ERK1/2 activation and total cell lysate and apical membrane NHE-3 abundance. The scrambled siRNA had no effect on ANG II endocytosis, ERK1/2 activation, or NHE-3 expression. These results suggest that AT(1) receptor-mediated endocytosis of extracellular ANG II may regulate proximal tubule sodium transport by increasing total and apical NHE-3 proteins. PMID- 17428841 TI - Human NADH:ubiquinone oxidoreductase deficiency: radical changes in mitochondrial morphology? AB - Malfunction of NADH:ubiquinone oxidoreductase or complex I (CI), the first and largest complex of the mitochondrial oxidative phosphorylation system, has been implicated in a wide variety of human disorders. To demonstrate a quantitative relationship between CI amount and activity and mitochondrial shape and cellular reactive oxygen species (ROS) levels, we recently combined native electrophoresis and confocal and video microscopy of dermal fibroblasts of healthy control subjects and children with isolated CI deficiency. Individual mitochondria appeared fragmented and/or less branched in patient fibroblasts with a severely reduced CI amount and activity (class I), whereas patient cells in which these latter parameters were only moderately reduced displayed a normal mitochondrial morphology (class II). Moreover, cellular ROS levels were significantly more increased in class I compared with class II cells. We propose a mechanism in which a mutation-induced decrease in the cellular amount and activity of CI leads to enhanced ROS levels, which, in turn, induce mitochondrial fragmentation when not appropriately counterbalanced by the cell's antioxidant defense systems. PMID- 17428842 TI - Olfactory epithelia exhibit progressive functional and morphological defects in CF mice. AB - In normal nasal epithelium, the olfactory receptor neurons (ORNs) are continuously replaced through the differentiation of progenitor cells. The olfactory epithelium (OE) of the cystic fibrosis (CF) mouse appears normal at birth, yet by 6 mo of age, a marked dysmorphology of sustentacular cells and a dramatic reduction in olfactory receptor neurons are evident. Electroolfactograms revealed that the odor-evoked response in 30-day-old CF mice was reduced approximately 45%; in older CF mice, a approximately 70% reduction was observed compared with the wild type (WT) response. Consistent with studies of CF airway epithelia, Ussing chamber studies of OE isolated from CF mice showed a lack of forskolin-stimulated Cl(-) secretion and an approximately 12-fold increase in amiloride-sensitive sodium absorption compared with WT mice. We hypothesize that the marked hyperabsorption of Na(+), most likely by olfactory sustentacular cells, leads to desiccation of the surface layer in which the sensory cilia reside, followed by degeneration of the ORNs. The CF mouse thus provides a novel model to examine the mechanisms of disease-associated loss of olfactory function. PMID- 17428843 TI - Plasma gelsolin: in search of its raison d'etre. Focus on "Modifications of cellular responses to lysophosphatidic acid and platelet-activating factor by plasma gelsolin". PMID- 17428844 TI - Hypotonicity causes actin reorganization and recruitment of the actin-binding ERM protein moesin in membrane protrusions in collecting duct principal cells. AB - Hypotonicity-induced cell swelling is characterized by a modification in cell architecture associated with actin cytoskeleton remodeling. The ezrin/radixin/moesin (ERM) family proteins are important signal transducers during actin reorganization regulated by the monomeric G proteins of the Rho family. We report here that in collecting duct CD8 cells hypotonicity-induced cell swelling resulted in deep actin reorganization, consisting of loss of stress fibers and formation of F-actin patches in membrane protrusions where the ERM protein moesin was recruited. Cell swelling increased the interaction between actin and moesin and induced the transition of moesin from an oligomeric to a monomeric functional conformation, characterized by both the COOH- and NH(2) terminal domains being exposed. In this conformation, which is stabilized by phosphorylation of a conserved threonine in the COOH-terminal domain by PKC or Rho kinase, moesin can bind interacting proteins. Interestingly, hypotonic stress increased the amount of threonine-phosphorylated moesin, which was prevented by the PKC-alpha inhibitor Go-6976 (50 nM). In contrast, the Rho kinase inhibitor Y 27632 (1 microM) did not affect the hypotonicity-induced increase in phosphorylated moesin. The present data represent the first evidence that hypotonicity-induced actin remodeling is associated with phosphorylated moesin recruitment at the cell border and interaction with actin. PMID- 17428845 TI - Emergence of mammalian cell-adapted vesicular stomatitis virus from persistent infections of insect vector cells. AB - Arboviruses (arthropod-borne viruses) represent quintessential generalists, with the ability to infect and perform well in multiple hosts. However, antagonistic pleiotropy imposed a cost during the adaptation to persistent replication of vesicular stomatitis virus in sand fly cells and resulted in strains that initially replicated poorly in hamster cells, even when the virus was allowed to replicate periodically in the latter. Once a debilitated strain started replicating continuously in mammalian cells, fitness increased significantly. Fitness recovery did not entail back mutations or compensatory mutations, but instead, we observed the replacement of persistence-adapted genomes by mammalian cell-adapted strains with a full set of new, unrelated sequence changes. These mammalian cell-adapted genomes were present at low frequencies in the populations with a history of persistence for up to a year and quickly became dominant during mammalian infection, but coexistence was not stable in the long term. Periodic acute replication in mammalian cells likely contributed to extending the survival of minority genomes, but these genomes were also found in strictly persistent populations. PMID- 17428846 TI - Pocket factors are unlikely to play a major role in the life cycle of human rhinovirus. AB - Human rhinovirus 14 (HRV14) is a member of the rhinovirus genus, which belongs to the picornavirus family, which includes clinically and economically important members, such as poliovirus, foot-and-mouth disease virus, and endomyocarditis virus. Capsid stability plays an important role in the viral infection process, in that it needs to be stable enough to move from cell to cell and yet be able to release its genetic material upon the appropriate environmental cues from the host cell. It has been suggested that certain host cell molecules, "pocket factors," bind to the WIN drug-binding cavity beneath the canyon floor and provide transient stability to a number of the picornaviruses. To directly test this hypothesis, HRV14 was mutated in (V1188M, C1199W, and V1188M/C1199W) and around (S1223G) the drug-binding pocket. Infectivity, limited proteolysis, and matrix-assisted laser desorption ionization analyses indicate that filling the drug-binding pocket with bulky side chains is not deleterious to the viral life cycle and lends some stabilization to the capsid. In contrast, studies with the S1223G mutant suggest that this mutation at least partially overcomes WIN drug mediated inhibition of cell attachment and capsid breathing. Finally, HRV16, which is inherently more stable than HRV14 in a number of respects, was found to "breathe" only at 37 degrees C and did not tolerate stabilizing mutations in the drug-binding cavity. These results suggest that it is the drug-binding cavity itself and not the putative pocket factor that is crucial for the capsid dynamics, which is, in turn, necessary for infection. PMID- 17428847 TI - Cytidine deaminases APOBEC3G and APOBEC3F interact with human immunodeficiency virus type 1 integrase and inhibit proviral DNA formation. AB - APOBEC3G (A3G) is a single-stranded DNA cytidine deaminase that targets retroviral minus-strand DNA and has potent antiviral activity against diverse retroviruses. However, the mechanisms of A3G antiviral functions are incompletely understood. Here we demonstrate that A3G, A3F, and, to a lesser extent, the noncatalytic A3GC291S block human immunodeficiency virus type 1 (HIV-1) replication by interfering with proviral DNA formation. In HIV-1 virions, A3G interacted with HIV-1 integrase and nucleocapsid, key viral factors for reverse transcription and integration. Unlike A3G, the weak antiviral A3C cytidine deaminase did not interact with either of these factors and did not affect viral reverse transcription or proviral DNA formation. Thus, multiple steps of the HIV 1 replication cycle, most noticeably the formation of proviral DNA, are inhibited by both cytidine deamination-dependent and -independent mechanisms. PMID- 17428848 TI - Epidermal growth factor receptor is not required for human cytomegalovirus entry or signaling. AB - Human cytomegalovirus (HCMV) can bind, fuse, and initiate gene expression in a diverse range of vertebrate cell types. This broad cellular tropism suggests that multiple receptors and/or universally distributed receptors mediate HCMV entry. Our laboratory has recently discovered that certain beta1 and beta3 integrin heterodimers are critical mediators of HCMV entry into permissive fibroblasts (A. L. Feire, H. Koss, and T. Compton, Proc. Natl. Acad. Sci. USA 101:15470-15475, 2004). It has also been reported that epidermal growth factor receptor (EGFR) is necessary for HCMV-mediated signaling and entry (X. Wang, S. M. Huong, M. L. Chiu, N. Raab-Traub, and E. E. Huang, Nature 424:456-461, 2003). Integrins are known to signal synergistically with growth factor receptors, and this coordination was recently reported for EGFR and beta3 integrins in the context of HCMV entry (X. Wang, D. Y. Huang, S. M. Huong, and E. S. Huang, Nat. Med. 11:515 521, 2005). However, EGFR-negative cell lines, such as hematopoietic cells, are known to be infected by HCMV. Therefore, we wished to confirm a role for EGFR in HCMV entry and then examine any interaction between beta1 integrins and EGFR during the entry process. Surprisingly, we were unable to detect any role for EGFR in the process of HCMV entry into fibroblast, epithelial, or endothelial cell lines. Additionally, HCMV did not activate the EGFR kinase in fibroblast cell lines. We first examined HCMV entry into two EGFR-positive or -negative cell lines but observed no increase in entry when EGFR was expressed to high levels. Physically blocking EGFR with a neutralizing antibody in fibroblast, epithelial, or endothelial cell lines or blocking EGFR kinase signaling with a chemical inhibitor in fibroblast cells did not inhibit virus entry. Lastly, we were unable to detect phosphorylation of EGFR in fibroblasts cells in response to HCMV stimulation. Our findings demonstrate that EGFR does not play a significant role in HCMV entry or signaling. These results suggest that specific integrin heterodimers either act alone as the primary entry receptors or interact in conjunction with an additional receptor(s), other than EGFR, to facilitate virus entry. PMID- 17428849 TI - The host protein Staufen1 participates in human immunodeficiency virus type 1 assembly in live cells by influencing pr55Gag multimerization. AB - Human immunodeficiency virus type 1 (HIV-1) requires the sequential activities of virus-encoded proteins during replication. The activities of several host cell proteins and machineries are also critical to the completion of virus assembly and the release of infectious virus particles from cells. One of these proteins, the double-stranded RNA-binding protein Staufen1 (Stau1), selectively associates with the HIV-1 genomic RNA and the viral precursor Gag protein, pr55Gag. In this report, we tested whether Stau1 modulates pr55Gag assembly using a new and specific pr55Gag oligomerization assay based on bioluminescence resonance energy transfer (BRET) in both live cells and extracts after cell fractionation. Our results show that both the overexpression and knockdown of Stau1 increase the pr55Gag-pr55Gag BRET levels, suggesting a role for Stau1 in regulating pr55Gag oligomerization during assembly. This effect of Stau1 on pr55Gag oligomerization was observed only in membranes, a cellular compartment in which pr55Gag assembly primarily occurs. Consistently, expression of Stau1 harboring a vSrc myristylation signal led to a 6.5-fold enrichment of Stau1 in membranes and a corresponding enhancement in the Stau1-mediated effect on pr55Gag-pr55Gag BRET, demonstrating that Stau1 acts on assembly when targeted to membranes. A role for Stau1 in the formation of particles is further supported by the detection of membrane-associated detergent-resistant pr55Gag complexes and the increase of virus-like particle release when Stau1 expression levels are modulated. Our results indicate that Stau1 influences HIV-1 assembly by modulating pr55Gag pr55Gag interactions, as shown in a live cell interaction assay. This likely occurs when Stau1 interacts with membrane-associated assembly intermediates.